PMID- 3024096 TI - Test and teach. Number fifty-three. Diagnosis: Fibrolamellar carcinoma of the liver. PMID- 3024097 TI - Foot deformities in infants and children. AB - Foot deformities may reflect a generalized disorder, especially a neurologic problem; thus, the child should have a brief general examination. Many infantile foot deformities, such as calcaneovalgus, are postural and self-correcting. Metatarsus varus is not referred for treatment until age 2 months and then only if the deformity is moderate or severe. Fixed forefoot equinus and heel varus characterize a clubfoot, which requires immediate treatment. Corrective shoes are not advised as the primary treatment for metatarsus varus or clubfoot but often are prescribed to maintain the corrected position after serial casts. Flexible flatfoot is a manifestation of a constitutional laxity affecting all ligaments and joints. The feet appear abnormal because of weight-bearing stresses. Most children with flatfoot achieve a partial correction spontaneously. Current research does not document that treatment with corrective shoes or inserts produces a result better than the partial correction that occurs naturally. PMID- 3024098 TI - Microcephaly and congenital cytomegalovirus infection: a combined prospective and retrospective study of a Swedish infant population. AB - Microcephaly and its etiology were studied in an unselected Swedish urban infant population. Virtually, all live-born infants (14,724) born between October 1977 and December 1983 in the city of Malmo, Sweden, were included in the study. Special attention was given to the role of congenital infections, particularly to cytomegalovirus infection. The infant population was studied from two points of view. One part of the study was prospective and based on regular cytomegalovirus isolation in urine within the first week of life. About 80% of the newborns were adequately studied by this test. None of 56 infants shown to be cytomegalovirus excreters (congenitally infected) and followed up were born with or developed microcephaly (head circumference smaller than 3 SD below the mean for age and sex) during the first 1 to 7 years of life. However, two of the 56 infants had a head circumference of -2 SD. In the beginning of 1985, an inventory was made of the presence of symptomatic microcephaly in the above mentioned population still living in the city or deceased there. Of about 10,000 such children, 12 were found to have symptomatic microcephaly. By studies of personal, clinical, and laboratory data and by retrospective serologic studies of frozen pre- and postconceptional maternal sera, a possible explanation or a recognized syndrome was obtained in ten of the 12 cases. In one of them, the mother had a primary cytomegalovirus infection, possibly in early pregnancy. Although the infant had symptoms compatible with a congenital infection, no laboratory evidence of transmitted infection was found. In no case were congenital rubella virus or Toxoplasma gondii infections suspected. PMID- 3024099 TI - A comparison of miniature end-plate potentials at normal, denervated, and long term botulinum toxin type A poisoned frog neuromuscular junctions. AB - The effects of denervation and long-term botulinum toxin type A (BoTx) poisoning on miniature end-plate potentials (m.e.p.p.s) in the frog were studied with intracellular microelectrode recording. BoTx reduced the frequency of m.e.p.p.s to less than 1% of the level seen in untreated frogs, leaving a large percentage of tiny m.e.p.p.s and slow-rising m.e.p.p.s (slow m.e.p.p.s). Unlike what is observed in the rat, the frequency of slow m.e.p.p.s never increased above the low rate measured in the untreated controls, and in fact slightly but significantly decreased after BoTx. A comparison of the m.e.p.p.s seen after BoTx poisoning (BoTx m.e.p.p.s) and m.e.p.p.s seen after denervation (Schwann m.e.p.p.s) revealed many similarities between the two including amplitude and time-to-peak distributions, temperature Q10 values and responses to several drugs and procedures. However, it was concluded that BoTx m.e.p.p.s do not originate from the Schwann cells because denervation of BoTx-paralysed frogs abolishes all m.e.p.p.s and the drug 4-aminoquinoline affects BoTx m.e.p.p.s and Schwann m.e.p.p.s in opposite ways, increasing the frequency of the former while almost eliminating the latter. BoTx m.e.p.p.s and Schwann m.e.p.p.s probably represent similar processes of secretion which are non-specific in nature, having a lower energy barrier than for normal release and not originating from specialized areas of transmitter release. PMID- 3024100 TI - [Transcatheter therapy of unresectable hepatocellular carcinoma--analysis of prognostic factors and predicted prognoses by multivariate analysis]. PMID- 3024101 TI - Insertion elements and transitions in cloned mouse mammary tumour virus DNA: further delineation of the poison sequences. AB - The provirus of mouse mammary tumour virus (MMTV) is reputed to contain sequences within the viral gag gene that prevent or inhibit its propagation as a recombinant DNA clone in Escherichia coli. Here we report the successful isolation of several lambda and plasmid clones comprising the 5' virus-host DNA junction fragments from integrated MMTV proviruses in BR6 mice. Although the lambda clones appeared intact, almost all of the plasmids were found to contain the bacterial insertion sequences IS1 or IS2 within a small region of the gag gene. One nondisrupted clone was recovered which had undergone multiple G to A transitions, some of which created stop codons in gag. These results have provided more precise information as to the location of the poison sequences and are discussed in relation to possible explanations for the phenomenon. PMID- 3024102 TI - The control of herpes simplex virus type-1 late gene transcription: a 'TATA box'/cap site region is sufficient for fully efficient regulated activity. AB - The transcriptional programme of herpes simplex virus type 1 (HSV-1) is organised into three principle phases; immediate-early (IE), early (E) and late. The appearance of IE gene products provides the switch for E transcription. Abundant expression of late genes requires viral DNA replication. There is some overlap between E and late genes according to their degree of dependence on DNA replication. The pattern of expression of gene US11 is regulated with 'true-late' kinetics (Johnson et al., 1986). In a transient assay system, regulation of a plasmid-borne US11 promoter mimics its viral counterpart, and has a similar dependence on DNA replication for abundant expression. Using plasmids which contain a functional HSV-1 origin of replication (ORIS), we have identified the sequence requirements for the expression of late genes. All DNA sequence elements necessary for fully efficient regulated expression of US11 lie within 31 bp of the RNA cap sites; therefore it appears that a late gene promoter consists only of a proximal 'TATA-box' and cap-site region. We tested this hypothesis by removing the distal upstream region of the gD promoter (which is required for its normal regulation as an early promoter) and linking this truncated promoter to ORIS. This resulted in the conversion of gD promoter regulation to late gene kinetics during virus superinfection. The implications of these results for the mechanisms of HSV gene regulation are discussed. PMID- 3024103 TI - DNA amplification--deletion in a spontaneous mutation of the hamster aprt locus: structure and sequence of the novel joint. AB - In a collection of spontaneous mutants of Chinese hamster ovary cells selected for deficiency in adenine phosphoribosyl transferase (aprt) activity, one was detected having not only a deletion of aprt coding sequences but also an apparent amplification of remaining sequences. The HindIII fragment bearing the novel joint was cloned and sequenced revealing a complex gene rearrangement. A deletion of at least 9 kb extending upstream from the aprt locus is accompanied by an inverted duplication of flanking sequences 672 bp downstream from the novel joint. This unit is amplified three to four times with the net result of some sequences being increased as much as eight fold in copy number because of the duplication. The fidelity of the sequences involved is preserved. We propose a model which could account for this inverted duplication. PMID- 3024104 TI - Genomic localization of hepatitis B virus in a human hepatoma cell line. AB - The integration of hepatitis B viral sequences in the human hepatoma Alexander cell line has been investigated after fractionation of the cell line DNA by centrifugation in a Cs2SO4/BAMD (3,6-(bis-acetato mercurimethyl) dioxane) density gradient. Eight out of nine integrated viral sequences were localized in DNA component H3, which only represents 4% of the human genome and matches the base composition of HBV sequences. These results indicate a targeting and/or a higher stability of the latter in a specific, small compartment of the host genome. PMID- 3024105 TI - The alpha-amylase gene in Drosophila melanogaster: nucleotide sequence, gene structure and expression motifs. AB - We present the complete nucleotide sequence of a Drosophila alpha-amylase gene and its flanking regions, as determined by cDNA and genomic sequence analysis. This gene, unlike its mammalian counterparts, contains no introns. Nevertheless the insect and mammalian genes share extensive nucleotide similarity and the insect protein contains the four amino acid sequence blocks common to all alpha amylases. In Drosophila melanogaster, there are two closely-linked copies of the alpha-amylase gene and they are divergently transcribed. In the 5'-regions of the two gene-copies we find high sequence divergence, yet the typical eukaryotic gene expression motifs have been maintained. The 5'-terminus of the alpha-amylase mRNA, as determined by primer extension analysis, maps to a characteristic Drosophila sequence motif. Additional conserved elements upstream of both genes may also be involved in amylase gene expression which is known to be under complex controls that include glucose repression. PMID- 3024106 TI - The mRNA and RNA-copy pseudogenes encoding TM30nm, a human cytoskeletal tropomyosin. AB - We have determined the sequence of a 2.5 kb mRNA in human fibroblasts encoding a 248 amino acid cytoskeletal tropomyosin. The protein product of this mRNA is TM30nm, one of five tropomyosin-like proteins in human fibroblasts. The structural gene encoding this mRNA can also produce a 1.3 kb mRNA encoding a 285 amino acid skeletal muscle alpha-tropomyosin by tissue-specific alternative mRNA splicing. However, the multiple RNA-copy pseudogenes of this gene family are derived largely if not exclusively from transcripts processed according to the pattern observed in non-muscle cells. PMID- 3024107 TI - Splice site consensus sequences are preferentially accessible to nucleases in isolated adenovirus RNA. AB - The conformation of RNA sequences spanning five 3' splice sites and two 5' splice sites in adenovirus mRNA was probed by partial digestion with single-strand specific nucleases. Although cleavage of nucleotides near both 3' and 5' splice sites was observed, most striking was the preferential digestion of sequences near the 3' splice site. At each 3' splice site a region of very strong cleavage is observed at low concentrations of enzyme near the splice site consensus sequence or the upstream branch point consensus sequence. Additional sites of moderately strong cutting near the branch point consensus sequence were observed in those sequences where the splice site was the preferred target. Since recognition of the 3' splice site and branch site appear to be early events in mRNA splicing these observations may indicate that the local conformation of the splice site sequences may play a direct or indirect role in enhancing the accessibility of sequences important for splicing. PMID- 3024108 TI - The purification of the Escherichia coli UvrABC incision system. AB - The UvrA, UvrB and UvrC proteins of Escherichia coli have been purified in good yields to homogeneity with rapid three- or four-step purification procedures. The cloned uvrA and uvrB genes were placed under control of the E. coli bacteriophage lambda PL promoter for amplification of expression. Expression of the uvrC gene could not be amplified by this strategy, however, subcloning of this gene into the replication-defective plasmid pRLM24 led to significant overproduction of the UvrC protein. The purified UvrA protein, with its associated ATPase activity, has a molecular weight of 114,000, the purified UvrB is an 84,000 molecular weight protein and the UvrC protein has a molecular weight of 67,000. PMID- 3024109 TI - The effect of Escherichia coli Uvr protein binding on the topology of supercoiled DNA. AB - The effects of the binding of the E. coli UvrA and UvrB proteins on the linking number (delta L) of superhelical DNA has been measured. The effects of cofactor ATP structure on UvrAB-nucleoprotein complex formation revealed that nucleotide binding, not hydrolysis, is sufficient to locally unwind the DNA helix of both ultraviolet light-damaged as well as undamaged DNAs. The extent of this unwinding is of the same order of magnitude as the nucleotide distances of the double incision sites generated by the UvrABC endonucleolytic reaction. PMID- 3024110 TI - Nucleotide sequence of the cellulase gene celD encoding endoglucanase D of Clostridium thermocellum. AB - The nucleotide sequence of the celD gene, encoding the previously crystallized endoglucanase D of Clostridium thermocellum, is reported. The enzyme shares a conserved, reiterated domain with the COOH-terminal end of endoglucanases A and B from the same organism. The overexpression in Escherichia coli of celD subcloned in pUC8 appears to result from a translational fusion of the NH2-terminal end of the endoglucanase with the NH2-terminal end of beta-galactosidase. PMID- 3024111 TI - Efficient construction of cDNA libraries in plasmid expression vectors using an adaptor strategy. AB - We describe a method for the construction of large DNA fragment libraries in plasmid vectors, in which complementary, single-stranded extensions are ligated onto both vector and insert DNA using un-phosphorylated adaptor oligonucleotides. Special consideration has been taken of the requirements of expression screening as follows: cDNA synthesis using random oligonucleotide primers is described which maximises the probability of obtaining open reading frame fragments from large mRNA molecules, the adaptors use codons found in high abundance E. coli proteins to minimise problems of premature termination when using strong promoters, and the sequence encoded by the adaptors, when cloned into the bacterial expression vector pEX1, promotes a surface location for the foreign antigenic determinant where it is accessible to antibodies used for screening. PMID- 3024112 TI - Nucleotide sequence of the AAD(2'') aminoglycoside adenylyltransferase determinant aadB. Evolutionary relationship of this region with those surrounding aadA in R538-1 and dhfrII in R388. AB - The nucleotide sequence of the aadB gene which confers resistance to kanamycin, gentamicin, and tobramycin has been determined. The size of the longest reading frame is 747 bases encoding a protein of predicted size 27,992 daltons. A segment of the aadB gene sequence (including the promoter region) was found upstream of the aadA gene in R538-1 and of the dhfrII gene in R388 and the proposed promoters for these genes coincide with the aadB promoter region. The sequence homology extends upstream to the end of the sequenced regions of R388 and R538-1. Almost perfect homology was also found between the sequences 3'- to the aadB gene and 3' to the aadA genes of R538-1 and pSa. This segment includes a 59 base element previously found flanking the Tn7 aadA gene. A model is presented for the evolution of this region of the plasmid genomes in which the 59- base element functions as an insertional "hot spot" and the possibility that this region is analogous to the aadA/aadB region of the Tn21- like transposon family is considered. PMID- 3024113 TI - Nucleotide sequence and analysis of the 58.3 to 65.5-kb early region of bacteriophage T4. AB - The complete 7.2-kb nucleotide sequence from the 58.3 to 65.5-kb early region of bacteriophage T4 has been determined by Maxam and Gilbert sequencing. Computer analysis revealed at least 20 open reading frames (ORFs) within this sequence. All major ORFs are transcribed from the left strand, suggesting that they are expressed early during infection. Among the ORFs, we have identified the ipIII, ipII, denV and tk genes. The ORFs are very tightly spaced, even overlapping in some instances, and when ORF interspacing occurs, promoter-like sequences can be implicated. Several of the sequences preceding the ORFs, in particular those at ipIII, ipII, denV, and orf61.9, can potentially form stable stem-loop structures. PMID- 3024114 TI - The DNA replication origins of herpes simplex virus type 1 strain Angelotti. AB - The nucleotide sequences of the origins of DNA replication (ori) of the S- and L component (oriS, oriL) of the herpes simplex virus type 1 (HSV-1) standard genome were determined from HSV-1 strain Angelotti (ANG). In contrast to other HSV-1 strains, the ANG oriS sequence revealed an insertion of an TA-dinucleotide in an otherwise very conserved but imperfect palindromic sequence of 47 bp. The oriL sequence of the standard ANG genome was found to be identical to that of an ANG class II defective genome which exhibits a duplication of a 144 bp palindrome. A model is presented to explain the origination of the amplified ANG oriL sequences from the parental genome with a single copy of oriL via illegitimate recombination. Alignment of the ori sequences of HSV, adeno- and papovaviruses unveiled that the HSV ori region can be subdivided into two distinct sites of homology to the DNA initiation signals of papova- and adenoviruses, suggesting that the HSV origins of replication comprise elements for DNA replication by both, cellular and virus-encoded DNA polymerases. PMID- 3024116 TI - pG95 alpha 1-7dIII/RI, a single copy clone at Xp11.4 which recognises a TaqI polymorphism (DXS209). PMID- 3024115 TI - Conserved sequence motifs upstream from the co-ordinately expressed vitellogenin and apoVLDLII genes of chicken. AB - The vitellogenin and apoVLDLII yolk protein genes of chicken are transcribed in the liver upon estrogenization. To get information on putative regulatory elements, we compared more than 2 kb of their 5' flanking DNA sequences. Common sequence motifs were found in regions exhibiting estrogen-induced changes in chromatin structure. Stretches of alternating pyrimidines and purines of about 30 nucleotides long are present at roughly similar positions. A distinct box of sequence homology in the chicken genes also appears to be present at a similar position in front of the vitellogenin genes of Xenopus laevis, but is absent from the estrogen-responsive egg-white protein genes expressed in the oviduct. In front of the vitellogenin (position -595) and the VLDLII gene (position -548), a DNA element of about 300 base-pairs was found, which possesses structural characteristics of a mobile genetic element and bears homology to the transposon like Vi element of Xenopus laevis. PMID- 3024117 TI - Hypersensitive sites in the 5' and 3' flanking regions of the cysteine proteinase I gene of Dictyostelium discoideum. AB - The cysteine proteinase I gene of Dictyostelium discoideum is a developmentally regulated single copy gene. Specific sites in the 5' and the 3' flanking regions of the gene were cleaved by an endogenous nuclease when the gene was being transcribed. The majority of these sites were not cut when the gene was inactive. A dramatic change in the pattern of micrococcal nuclease and DNase I hypersensitive sites occurred in the 5' flanking region when transcription commenced at the 8 h stage of development. The major sites, doublets at -220/-300 bp and -670/-770 bp upstream of the transcription start site, corresponded to those cut by the endogenous nuclease. When transcription subsequently ceased the hypersensitive sites did not significantly change, indicating the gene remained in an activated state. The micrococcal nuclease hypersensitive sites in the 3' flanking region did not change significantly during development. PMID- 3024118 TI - Antibiotics which can alter the rotational orientation of nucleosome core DNA. AB - Four well-characterised DNA-binding ligands have been tested for effects on reconstituted nucleosome core particles containing the 160 bp tyrT DNA fragment. Two, netropsin and berenil, were found to change the rotational orientation of the DNA on the surface of the protein as judged by marked alterations in the pattern of fragments produced by exposure to DNAase I. Qualitatively their effects were very similar to those previously reported for the related antibiotic distamycin, suggesting that the phenomenon of induced rotation may be a characteristic property of ligands which bind in the narrow groove of the DNA helix. Two intercalators did not produce the effect but, at high concentrations, caused gross disruption of the nucleoprotein structure with apparent release of DNA from the histone octamer. At moderate concentrations little or no effect was detectable with nogalamycin, suggestive of failure to bind as a result of constraints on local opening of the DNA helix. With moderate concentrations of actinomycin, protection of GpC sequences was clearly visible together with some evidence of increase in helix pitch, but no sign of altered phasing of DNA within the nucleosome core particles. PMID- 3024119 TI - Model for alternative RNA processing in human calcitonin gene expression. AB - The alternative RNA processing pathways in human calcitonin gene (CALC-I gene) expression were investigated using steady state RNA isolated from human medullary thyroid carcinoma (MTC) and from a culture line derived from this tumor. On Northern blots the mature 1.0 kilobases (Kb) calcitonin (CT) - and 1.1 Kb calcitonin gene-related peptide (CGRP) mRNAs were detected with CALCI gene specific probes as well as high molecular weight poly (A) containing RNAs of 2.1, 2.3, 3.3, 4.2, 5.0 and 5.7 Kb. The 5.7 Kb RNA was identified as the poly(A) tailed primary transcript containing sequences corresponding to all 6 exons and 5 introns of the CALC-I gene. From the composition of the other RNAs the splicing order of the different introns could be deduced. The results suggest the following model. First all introns not involved in alternative processing (introns 1, 2 and 5) are spliced from the 5.7 Kb RNA in rapid successive reactions yielding a 3.3 Kb RNA, which accumulates. From this 3.3 Kb RNA, the last common intermediate in the alternative processing pathway, CT mRNA is formed by splicing of intron 3 and poly(A) addition at exon 4, in this order or the reverse order via 2.3 Kb or 2.1 Kb RNA intermediates respectively. Alternatively, the whole intron 3-exon 4-intron 4 region is spliced from the 3.3 Kb RNA yielding CGRP mRNA. The temporal sequence of poly(A) addition at exons 4 and 6 may relate to the observed structural differences between the poly(A) addition signals at these sites. The ratio of CT- to CGRP mRNA may relate also to the differences in the primary structures of the intron 3- and intron 4 splice acceptor sites. PMID- 3024120 TI - Gene organization of the small subunit of human calcium-activated neutral protease. AB - The gene for the small subunit of human calcium-activated neutral protease was isolated and sequenced. It is 11 kb long and comprises 11 exons. No TATA or CAT box was found upstream of the possible transcription initiation sites, but there are three so-called G-C box sequences and one G-C box-like sequence, which are usually found in "house-keeping" genes. The first exon (exon 1) contains only the 5'-noncoding sequence and exon 2 encodes the Gly-rich hydrophobic domain. Each of the four calcium-binding loop regions is encoded by one exon (exons 7-10). The intron breakpoints in the C-terminal calcium-binding domain (exons 4-11) completely coincide with those of the chicken large subunit gene. These findings suggest that the small and large subunits have evolved from the same ancestral calcium-binding protein and have retained the original gene organization. PMID- 3024121 TI - A chimeric mouse histone H4 gene containing either an intron or poly(A) addition signal behaves like a basal histone. AB - We have modified the basic structure of the mouse H4 histone gene by introducing, in one case, the IVS-II of the human beta globin gene in the middle of the H4 coding region and, in the second case, the poly(A) addition signal from either the chicken vimentin gene or the alpha globin gene, displacing the hairpin loop structure in the 3' direction. Constructs were placed into the vector, PSV2gpt, and stably transformed into L cells. Pools of 100-500 independent transformants were analyzed for H4 expression. Even though the intron is processed correctly, the growth regulated expression of the modified gene is lost and the gene is now expressed at a constant basal level. Furthermore, unprocessed transcripts accumulate in the nucleus of Go cells when compared to exponentially growing cultures. Polyadenylated H4 RNA is correctly processed but expressed at reduced levels (30 fold) in a constitutive manner, independent of the growth state of the cell. The altered expression of these chimeric H4 genes compared to the endogenous copy or the transfected wild type gene suggests a structural model to explain the cell cycle independent expression of the basal histones. PMID- 3024122 TI - A growth-responsive gene (16C8) in normal mouse fibroblasts homologous to a human collagenase inhibitor with erythroid-potentiating activity: evidence for inducible and constitutive transcripts. AB - We present the DNA sequence of an essentially full-length cDNA clone of 16C8, a growth factor-inducible gene isolated from a mouse embryo fibroblast cDNA library. The 0.9-kb mRNA encodes an Mr 22,500 protein that has substantial homology to a human protein with the reported abilities to potentiate erythroid differentiation and to inhibit collagenases and other tissue metalloproteinases. The N-terminus of the predicted protein has a hydrophobic nature characteristic of secreted proteins, and two potential sites for N-linked glycosylation are present. The cytoplasmic concentration of 16C8 mRNA is maximal in mid G1 at about 6 h after serum stimulation of quiescent fibroblasts. Northern blot analysis showed a progressive reduction in the size of the induced 16C8 transcripts with increasing time after serum stimulation. This was shown to be due to the reduction in length of the poly(A) tails. S1 analysis of the 5' portion of the mRNA revealed the presence of three different species of transcript, only one of which was inducible. PMID- 3024123 TI - Adipsin, the adipocyte serine protease: gene structure and control of expression by tumor necrosis factor. AB - We have isolated, mapped and sequenced adipsin, the adipocyte differentiation dependent serine protease gene. This gene, which is present in a single form in the mouse, spans 1.7 kilobases and contains five exons. While the basic exon structure characteristic of serine protease genes is conserved in adipsin, there is also a fusion of two exons that are separate in other serine proteases. The sequence data also suggests a mechanism of alternative splicing which appears to account for the generation of two adipsin mRNA species differing by only three nucleotides and encoding two different signal peptides. To investigate the control of adipsin expression we have examined the effects of tumor necrosis factor (TNF) on adipocytes. The level of adipsin RNA is dramatically decreased by hormone treatment, but the change occurs more slowly than for other fat cell mRNAs, such as glycerophosphate dehydrogenase. These results show that adipsin is a novel serine protease gene whose expression is regulated by a macrophage derived factor which modulates expression of other adipocyte-specific RNAs. PMID- 3024124 TI - Deletion and rearrangement of plasmid DNA during transformation of Escherichia coli with linear plasmid molecules. AB - When E. coli was transformed with linearized pBR322 DNA, many transformants contained recircularized plasmids bearing deletions and other rearrangements. Most aberrant molecules were less than monomeric length and had lost the restriction site used for linearization, with the deleted region extending mono- (type Ia) or bi-directionally (type Ib). Type II deletants were greater than monomeric but less than dimeric and contained the pBR322 sequence in direct repeat with deletion at one or both junctions (type IIa) or in inverted repeat with loss of sequence at both junctions (type IIb). Type III deletants were greater than dimeric but less than trimeric, consisting of pBR322 sequences in both direct and inverse repeat with deletions at two or more junctions. Transformation frequencies for linear DNA were drastically reduced in xth-1- bacteria with type IIb deletants predominating in transformants. This indicates that exonuclease III is important for perfect recyclization of plasmids and the generation of type I deletants. In vivo recyclization of in vitro ligation products explains many of the aberrant DNA molecules that are encountered during gene cloning. PMID- 3024125 TI - Altered DNA conformations in the gene regulatory region of torsionally-stressed SV40 DNA. AB - We used mung bean nuclease to probe the SV40 genome for DNA unwinding and unpairing. Cleavage occurred at a limited number of specific sites in supercoiled, but not relaxed DNA. The number and location of cleavage sites depended upon Mg2+ concentration. Without Mg2+, cutting occurred mainly in one early denaturation region located 3' to the t antigen gene and within the T antigen gene intron. With Mg2+, cleavage occurred at a number of alternative sites in the genome. Certain Mg2+ concentrations favored cleavage in the gene regulatory region. These cleavages were mapped at single nucleotide resolution and occurred in both transcriptional enhancers and upstream from the start of major late gene transcription. The cleavages occurred between 5 bp inverted repeat sequences, consistent with the recognition of unusually small cruciform structures. PMID- 3024126 TI - Sendai virus induces high levels of tumor necrosis factor mRNA in human peripheral blood leukocytes. AB - Sendai virus induces human peripheral blood leukocytes to produce high levels of tumor necrosis factor (TNF) mRNA. TNF mRNA can represent as much as 0.6% of the total mRNA. Kinetic studies indicate that the level of TNF mRNA peaks about 2 hours before that of IFN-alpha mRNA produced in the same system. Although the peak levels of TNF and IFN-alpha mRNA were similar, TNF in the culture supernatants was at a 200 fold lower level than IFN-alpha. Cloning and sequence analysis of TNF cDNA isolated from peripheral blood leukocytes RNA showed that normal human cells in response to Sendai virus produce TNF identical to that previously isolated and cloned from tumor-derived cell lines. A bacterial expression system was used to produce the cloned TNF at a maximum level of 2 X 10(6) units per ml of culture. PMID- 3024127 TI - 1731, a new retrotransposon with hormone modulated expression. AB - We report here the characterisation of 1731, a new copia-like element of Drosophila melanogaster. 1731 was first isolated in a screening for ecdysterone modulated genes. This element is about 4.6 Kb long and is flanked by two long terminal repeats (LTRs) 336 base pairs in length. The whole 1731 element is transcribed into polyA+ RNAs, and these transcripts decrease rapidly upon hormonal treatment. 1731 is moderately repeated in the fly genome and slightly amplified in Kc/cells where extrachromosomal circular forms are found. The LTRs were sequenced in one cloned copy of 1731 and show a structural organisation similar to that of several other copia-like elements and retroviral proviruses. Small nucleotide stretches, similar to those found in Mouse Mammary Tumor Virus LTRs and known to be important in its regulation by a steroid hormone, occur in 1731 LTRs. PMID- 3024128 TI - Site directed mutagenesis experiments suggest that Glu 111, Glu 144 and Arg 145 are essential for endonucleolytic activity of EcoRI. AB - We have constructed a plasmid (pRIF 309+) carrying the EcoRI restriction endonuclease gene and the f1 origin of replication. Upon transformation of this plasmid into E. coli and infection with bacteriophage f1 single stranded plasmids are produced which can be used for sequencing and site directed mutagenesis. Using this single stranded DNA and synthetic oligodeoxynucleotides we have introduced point mutations at defined positions of the EcoRI gene. Since in pRIF309+ the EcoRI gene is under the control of the pL-promoter, high level expression of the mutated EcoRI gene could be obtained upon induction. Mutant EcoRI enzymes were purified to homogeneity and characterized in structural and functional terms. Our results demonstrate that the Glu 111----Gln, Glu 144----Gln and Arg 145----Lys -mutants adopt a very similar conformation as the wild type enzyme, but have by two orders of magnitude smaller specific activities than the wild type enzyme, mainly due to a reduction of the Vmax-value. PMID- 3024129 TI - Cloning of cDNA for human T-cell replacing factor (interleukin-5) and comparison with the murine homologue. AB - We have cloned cDNA for T-cell replacing factor (interleukin-5), which replaces T cell helper function for normal B cells which secrete immunoglobulin, from human T cell leukemia line, ATL-2, using mouse interleukin-5 cDNA as probe. Total nucleotide sequence of the cDNA (816 base pairs) was determined and compared with that of mouse interleukin-5 cDNA. The cloned cDNA encoded the interleukin-5 precursor of 134 amino acids containing an N-terminal signal sequence. Although the human interleukin-5 precursor is one amino acid longer than the murine homologue, the sizes of the mature proteins appear similar. The nucleotide and amino acid sequence homologies of the coding regions of human and murine interleukin-5 are 77% and 70%, respectively. Human interleukin-5 synthesized by the direction of the cloned cDNA induced immunoglobulin synthesis in human B cells stimulated by Staphylococcus aureus mitogen. PMID- 3024130 TI - Structure of a Bacillus subtilis endo-beta-1,4-glucanase gene. AB - The nucleotide sequence of the portion of a Bacillus subtilis (strain PAP115) 3 kb Pst I fragment which contains an endo-beta-1, 4-glucanase gene has been determined. This gene encodes a protein of 499 amino acid residues (Mr = 55,234) with a typical B. subtilis signal peptide. Escherichia coli which has been transformed with this gene produces an extracellular endoglucanase with an amino terminus corresponding to the thirtieth encoded amino acid residue. The gene is preceded by a cryptic reading frame with a rho-independent terminator structure, and itself has such a structure in the immediate 3'-flanking region. We have also identified, in the 5'-flanking region, nucleotide sequences which resemble promoter elements recognized by Bacillus RNA polymerase E sigma 43. Comparison of the encoded amino acid sequence to other known beta-glucanases reveals a small region of similarity to the encoded protein of the Clostridium thermocellum celB gene. These similar regions may contain substrate-binding and/or catalytic sites. PMID- 3024131 TI - A simple and efficient enzymatic method for covalent attachment of DNA to cellulose. Application for hybridization-restriction analysis and for in vitro synthesis of DNA probes. AB - Single-stranded DNAs (ssDNAs) were covalently bound by a simple and efficient enzymatic method to a solid support matrix and used to develop several new procedures for gene analysis. The novel procedure to prepare a ssDNA stably coupled to a solid support employed T4 DNA ligase to link covalently oligo (dT) cellulose and (dA)-tailed DNA. Beginning with essentially any double stranded DNA the procedure generates a ssDNA linked by its 5' end to a cellulose matrix in a concentration of over 500 ng per mg. DNA from the plasmid pBR322 (4300 bp) and a fragment of the beta-globin gene (1800 bp) were coupled to the solid support and used for several experiments. The ssDNAs on the cellulose efficiently hybridized with as little as 5 pg of complementary double-stranded DNAs. The DNA hybrids formed on the solid support were specifically and efficiently cleaved by restriction endonucleases. These specific restriction cuts were utilized for the diagnosis of correct sequences. In addition, the ssDNA on the solid support served as an efficient template for the synthesis of complementary ssDNAs. The complementary synthesized ssDNAs were uniformly labeled, more than two kilobases in size, and largely full length. About 85% of the ssDNA linked to cellulose was available for the synthesis of complementary DNA, and after strand-separation, the preparation was reusable for the synthesis of additional complementary DNA. PMID- 3024132 TI - Nucleotide sequence of a Trypanosoma cruzi minicircle. PMID- 3024133 TI - Restriction mapping by preferential ligation of adjacent digestion fragments. PMID- 3024134 TI - [Radiotherapy of chemodectoma of the temporal bone area]. PMID- 3024135 TI - [Malignant fibrous histiocytoma of the palatine tonsil]. PMID- 3024136 TI - [Increased levels of alpha and beta subunits of chorionic gonadotropin in the serum of patients with primary lung cancer]. PMID- 3024137 TI - The spectrum of dental manifestations in vitamin D-resistant rickets: implications for management. PMID- 3024138 TI - Small cell carcinoma of the thyroid. A reclassification of cases originally diagnosed as small cell carcinomas of the thyroid. AB - 99 cases of undifferentiated carcinomas of the thyroid and nine cases of primary malignant Non-Hodgkin lymphomas of the thyroid were examined from 1967 to 1983 in our institute. Among the undifferentiated carcinomas nine cases were classified as small cell subtype. Over the years, the histopathological handling in regard to small cell subtype of undifferentiated carcinoma and primary malignant Non Hodgkin lymphoma has changed. The frequency of primary malignant Non-Hodgkin lymphoma has increased conspicuously in the last few years, whereas the number of small cell carcinomas decreased. A reclassification, based on immunohistochemical investigation for tumor markers of the nine cases originally diagnosed as small cell carcinomas from 1967 to 1983 revealed that five cases were poorly differentiated carcinomas or undifferentiated carcinomas of the spindle cell type. In three cases the primary diagnosis had to be revised into malignant Non Hodgkin lymphoma of the diffuse "histiocytic" type. The postmortem examination of the patient with the remaining small cell carcinoma "of the thyroid" revealed a clinically undetected small cell carcinoma of the lung with metastases to the cervical lymph nodes and the thyroid gland. These findings are in agreement with the results of several recently published papers indicating that true small cell carcinoma of the thyroid must be a very rare tumor. PMID- 3024139 TI - Light and electron microscopical morphometry of pituitary adenomas in hyperprolactinemia. AB - Two highly differentiated acidophil prolactin-cell adenomas with hyperprolactinemia (group I), 8 large cell chromophobe adenomas with hyperprolactinemia (group II), and 2 small cell chromophobe adenomas (group III), one of which was combined with hyperprolactinemia, were studied immunohistologically. Morphometry was performed on the light- and electron microscopical level. The 11 active adenomas were immunohistologically positive for prolactin, the 12th adenoma with normal prolactin plasma level was negative for prolactin. Light microscopical morphometry displayed significantly more cells of smaller size in the "small cell chromophobe" adenomas, whereas the large cell chromophobe adenomas and the highly differentiated prolactin cell adenomas were not different. Ultrastructural morphometry demonstrated significant differences between highly differentiated prolactin cell adenomas (group I), and large cell chromophobe adenomas (group II). The latter contain smaller "relative volumes" of nucleoli and of secretory granules, whereas the rough endoplasmic reticulum, the Golgi fields and the nuclei were not different. Comparison of large cell chromophobe adenomas (group II), and small cell chromophobe adenomas (group III) revealed significantly larger relative volumes of nuclei and of mitochondria but smaller volumes of rough endoplasmic reticulum and of Golgi fields in the small cell chromophobe adenomas. Significant differences between the active and the inactive adenoma of small cell chromophobe type in the group III were not found. In spite of the low quantity of small cell chromophobe adenomas and of acidophil prolactin cell adenomas, our data demonstrate that there exist distinct and significant light microscopical and ultrastructural differences between the three adenoma types.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3024140 TI - The spectrum of Epstein-Barr virus hepatitis in children. AB - The clinical and pathologic features of Epstein-Barr virus (EBV) hepatitis in 3 children are described. Manifestations included fever, hepatomegaly, disseminated intravascular coagulation, and failure of uptake of technetium by the reticuloendothelial system of the liver. Histologic features may mimic chronic active hepatitis and lymphoid malignancy. Two patients underwent exploratory laparotomy because of suspected tumor. Recognition of the wide spectrum of hepatic involvement in infectious mononucleosis is important in the differential diagnosis of hepatomegaly. Diagnosis should be made by measurement of IgM specific EBV antibodies. PMID- 3024141 TI - [Does asymmetric hamstring reflex indicate L5 radicular lesion?]. PMID- 3024142 TI - [Active principles of plant origin: a tool for studying membrane receptors]. AB - Ever since ancient times, considerable interest has been shown in plants for therapeutic use. Nowadays, however, studies of plant extracts are no longer based on empiricism; they provide great support to fundamental research, especially for a better understanding of the mediator/receptor couples. This applies to the study of cholinergic (atropine, muscarine, etc.), adrenergic (yohimbine, rauwolscine, etc.), dopaminergic (apomorphine, bromocriptine, etc.), purinergic (caffeine, theophylline, etc.), opiate (morphine), GABA (strychnine, muscimol, bicuculline, etc.), cardiac glycosides (gitaloxin, digitoxin) and PAF-acether receptors (ginkgolides from Ginkgo biloba). PMID- 3024143 TI - [Toxicity of oxygen, free radicals and defense mechanisms]. AB - Oxygen is essential for the life of aerobic organisms as the terminal acceptor of electrons. Due to its high affinity for lone electrons it fosters radicular reactions and opposes recombinations of radicals after rupture of covalent linkages. Lipoperoxidation processes then occur, attacking the cellular membranes. These radicular reactions are at the origin of many pathological phenomena. A complex arsenal of endogenous protective agents is normally present in all cells to face up to the danger of lone electron structures in oxygenated media. Food and natural or synthetic pharmacological agents compensate for the deficiencies. PMID- 3024144 TI - [Effect of Ginkgo biloba extract on the hemato-encephalic barrier]. AB - The different methods used to explore the blood-brain barrier (made up of cerebral capillary vessels), and notably, at molecular level, isolated microvessel preparations, have greatly improved our knowledge in this particular field. Some of these methods could be used to evaluate the protective effects of therapeutic substances, such as Ginkgo biloba extract, on the blood-brain barrier. PMID- 3024145 TI - [Treatment of the disorders of aging with Ginkgo biloba extract. From pharmacology to clinical medicine]. AB - Ginkgo biloba extract is prescribed in psychic and behavioural disorders of the elderly, in peripheral vascular deficiency and in functional disorders of ischaemic origin in the E.N.T. and eye areas. Numerous controlled clinical trials justify these prescriptions and are in agreement with the pharmacological data currently available. Experimentally, Ginkgo biloba extract has proved active on the circulatory and rheological functions, on neuronal metabolism threatened by ischaemia or hypoxia, on neurotransmission and on membrane lesions caused by free oxygenated radicals. Concerning Alzheimer's disease and dementia, no firm conclusion can be drawn for the time being due to the lack of animal model. However, experimental data suggest that the product may act on a number of major elements of these diseases. From what is already known about Ginkgo biloba extract, it appears that it fulfills the conditions laid down by the W.H.O. concerning the development of drugs effective against cerebral ageing. PMID- 3024146 TI - [Endocrinology at the time of hormonal receptivity]. PMID- 3024147 TI - [Cellular disorders induced by ischemia. The effect of trimetazidine]. AB - In contrast with the classical anti-anginal drugs, trimetazidine appears to be a very specific treatment, especially to the ischaemic cell. Whereas beta-blockers, nitrates and calcium antagonists all act outside the real ischaemic area (on peripheral veins or arteries, coronary vessels, whole heart muscle contractility, sinus node, endo-epicardial blood flow ratio, etc...) trimetazidine is only efficient on the ischaemia-induced loss of membrane functions and its consequences. The disturbance of tissue oxygen supply during ischaemia decreases mitochondrial ATP production and increases the generation of free radicals (O2-., OH-.). By diminishing the bioavailability of free radicals, trimetazidine lessens all their toxic effects: trimetazidine acts on the inactivation of enzymatic membrane proteins (which induces ATP over-reduction, creatine phosphokinase and lactate release outside the cell and electrolyte shifts); trimetazidine corrects the elevation of passive membrane permeability (increased by free radical-induced peroxidation of unsaturated membrane lipids); it antagonises free radical-induced stimulations of phospholipase A2 and thromboxane synthetase. In conclusion, trimetazidine restores energy-producing processes in the ischaemic heart cell by lessening the toxic effects of oxygenated free radicals. PMID- 3024148 TI - [Effect of trimetazidine on membrane changes induced by oxygen free radicals in human red cells]. AB - It is well known that oxygen free radicals are generated in different ischaemic tissues and can mediate cell injury. Furthermore, a loss of intracellular potassium is involved in heart ischemia. We previously developed a method to study a possible connection between these two phenomena in human erythrocytes treated in vitro with an oxygen free radical generator: phenazine methosulfate. We showed that transport systems specific to potassium are poorly inhibited (sodium-potassium pump; sodium-potassium cotransport) or unaffected (Gardos effect, chlorine-dependent potassium transport); that the erythrocyte potassium loss is essentially due to an increase in passive potassium permeability resulting from lipid peroxidation and that alterations of transport pathways are enhanced by diethyldithiocarbamate, a superoxide dismutase inhibitor. On the other hand, a cardioprotective drug, trimetazidine, indicated in ischaemic heart disease, which has no calcium antagonist action or coronary vasodilator effect, was studied as a possible candidate for preventing such damage by free radicals. Red cells collected from healthy donors previously treated with trimetazidine were incubated in vitro with phenazine methosulfate and diethyldithiocarbamate. We observed significant decreases in oxygen free radical-dependent passive potassium permeability and lipid peroxidation. This strongly suggests that trimetazidine possesses a free radical scavenger activity which may explain its cardioprotective role. PMID- 3024149 TI - [Monoclonal antibodies and new markers in breast oncology]. PMID- 3024150 TI - [Effect of hydrocortisone on the density-gradient distribution of autoreactive and 5'-nucleotidase-separating thymocytes]. AB - Rat thymocytes forming rosettes with autologous erythrocytes, are separated by centrifugation in the ficoll-verografin density gradient into two unequal fractions. A minor part of rosette forming cells is found in the gradient layers with the density of 1.069 and 1.073, a major part in the layers with the density of 1.090 and 1.080. After the administration of hydrocortisone to rats (2.5 mg per 100 g of mass) the number of rosette forming cells increased but in the gradient light fraction (d = 1.069). At 37 degrees C this thymocyte fraction separates the greatest amount of protein with 5'-nucleotidase activity. Under hydrocortisone influence the number of cells in this fraction and the amount of separable protein per 10(6) cells as well as 5'-nucleotidase activity per 10(6) increased in this fraction only. It has been assumed that an increase in 5' nucleotidase separated at 37 degrees C, is a more constant feature of cortisone resistant thymocytes than a low affinity to autologous erythrocytes which is usually regarded as a feature of mature thymocytes. PMID- 3024151 TI - Cloning and sequence analysis of rat bone sialoprotein (osteopontin) cDNA reveals an Arg-Gly-Asp cell-binding sequence. AB - The primary structure of a bone-specific sialoprotein was deduced from cloned cDNA. One of the cDNA clones isolated from a rat osteosarcoma (ROS 17/2.8) phage lambda gt11 library had a 1473-base-pair-long insert that encoded a protein with 317 amino acid residues. This cDNA clone appears to represent the complete coding region of sialoprotein mRNA, including a putative AUG initiation codon and a signal peptide sequence. The amino acid sequence deduced from the cDNA contains several Ser-Xaa-Glu sequences, possibly representing attachment points for O glycosidically linked oligosaccharides and one Asn-Xaa-Ser sequence representing a likely site for the N-glycosidically linked oligosaccharide. An interesting observation is the Gly-Arg-Gly-Asp-Ser sequence, which is identical to the cell binding sequence identified in fibronectin. The presence of this sequence prompted us to investigate the cell-binding properties of sialoprotein. The ROS 17/2.8 cells attached and attained a spread morphology on surfaces coated with sialoprotein. We could demonstrate that synthetic Arg-Gly-Asp-containing peptides efficiently inhibited the attachment of cells to sialoprotein-coated substrates. The results show that the Arg-Gly-Asp sequence also confers cell-binding properties on bone-specific sialoprotein. To better reflect the potential function of bone sialoprotein--we propose the name "osteopontin" for this protein. PMID- 3024152 TI - Characterization of a thrombomodulin cDNA reveals structural similarity to the low density lipoprotein receptor. AB - We have isolated a partial-length cDNA for bovine thrombomodulin from a lambda gt11 bovine adrenal capillary endothelial cell expression library. This was accomplished by immunoscreening with rabbit anti-thrombomodulin IgG heteroantibody and then rescreening with the initial positive recombinant insert. The cDNA obtained was authenticated by showing that it coded for the primary structure of two separate regions of bovine thrombomodulin. The nucleotide sequence of the largest cDNA allowed us to establish the structure of about 80% of the mature thrombomodulin transcript, which encodes the C-terminal half of the polypeptide. This membrane component is structurally similar to coated-pit receptors and is organized into domains that resemble those of the low density lipoprotein receptor. PMID- 3024153 TI - Identification of a domain within the phosphoprotein of vesicular stomatitis virus that is essential for transcription in vitro. AB - A full-length cDNA copy of the phosphoprotein (NS) mRNA of vesicular stomatitis virus (New Jersey serotype) was inserted into pGEM4 vector downstream of the promoter for bacteriophage SP6 RNA polymerase. Transcription of the cDNA in vitro resulted in the synthesis of NS mRNA, which was subsequently translated into NS protein in a cell-free rabbit reticulocyte system. The biological activity of the expressed NS protein was demonstrated by in vitro synthesis of mRNA by transcription-reconstitution with purified viral L protein and N-RNA template. Deletion mapping of the NS gene defined a specific domain between amino acid residues 213 and 247, which was essential for in vitro transcription. Removal of the COOH-terminal 21 amino acids, on the other hand, did not have a significant effect on transcription. This domain appears to be involved in efficient binding of NS protein to the N protein-RNA template. PMID- 3024154 TI - Human cDNA clones for four species of G alpha s signal transduction protein. AB - lambda gt11 cDNA libraries derived from human brain were screened with oligonucleotide probes for recombinants that code for alpha subunits of G signal transduction proteins. Eleven alpha s clones were detected with both probes and characterized. Four types of alpha s cDNA were cloned that differ in nucleotide sequence in the region that corresponds to amino acid residues 71-88. The clones differ in the codon for alpha s amino acid residue 71 (glutamic acid or aspartic acid), the presence or absence of codons for the next 15 amino acid residues, and the presence or absence of an adjacent serine residue. S1 nuclease protection experiments revealed at least two forms of alpha s mRNA. A mechanism for generating four species of alpha s mRNA by alternative splicing of precursor RNA is proposed. PMID- 3024155 TI - Glucocorticoid inhibition of transcription from episomal proopiomelanocortin gene promoter. AB - Glucocorticoid hormones alter transcription of specific genes. Glucocorticoid stimulated genes have been especially useful in unraveling molecular events responsible for positive gene regulation in mammals. The gene encoding proopiomelanocortin (POMC), which is under feedback inhibition by glucocorticoids, provides a model system to study negative gene regulation. Using an episomal bovine papilloma virus vector, we now demonstrate that a 769-base pair fragment containing the rat POMC promoter is sufficient to confer glucocorticoid inhibition. Transcription from the episomal POMC promoter starts at the same site and is inhibited by glucocorticoids to the same extent as POMC transcription in the anterior pituitary. Glucocorticoid inhibition is specific for POMC transcripts; neither bovine papilloma virus nor cellular actin mRNAs are affected by glucocorticoids. Thus, the episomal bovine papilloma virus/POMC system can be used to study the relationship between negative regulation of POMC transcription and chromatin structure. PMID- 3024156 TI - Topoisomerase I mutants: the gene on pBR322 that encodes resistance to tetracycline affects plasmid DNA supercoiling. AB - Plasmid pBR322 DNA isolated from topoisomerase I mutants of Escherichia coli and Salmonella typhimurium exhibits a distinctive supercoiling distribution characterized by an extremely heterogeneous distribution of linking numbers that contains highly negatively supercoiled topoisomers. Analysis of the supercoiling distributions of deletion and insertion derivatives of pBR322 shows that the presence of the gene on pBR322 encoding resistance to tetracycline is responsible for the unusual supercoiling distribution. Both an intact promoter and a portion of the remainder of the gene, but not the gene product, are required. However, no particular section of the gene outside the promoter appears to be necessary; only the size of the section remaining appears to be important. These observations suggest that transcription of this gene may be responsible for its effect on DNA supercoiling. PMID- 3024157 TI - Human cholesterol side-chain cleavage enzyme, P450scc: cDNA cloning, assignment of the gene to chromosome 15, and expression in the placenta. AB - Conversion of cholesterol to pregnenolone is mediated by P450scc [cholesterol, reduced-adrenal-ferrodoxin: oxygen oxidoreductase (side-chain-cleaving), EC 1.14.15.67]. RNA from several human adrenal samples was translated in vitro and immunoprecipitated with anti-bovine P450scc, indicating that P450scc mRNA represents about 0.5% of human adrenal mRNA in normal, hypertrophied, and malignant adrenals. A 1626-base-pair human adrenal P450scc cDNA was cloned in bacteriophage lambda gt10. Primer extension data indicated P450scc mRNA is about 1850 bases long and that all adrenal P450scc mRNA has the same 5' end. A full length clone containing 1821 bases was obtained from a human testis cDNA library to yield the complete sequence. The encoded human preP450scc contains 521 amino acids with a molecular weight of 60189.65. The testis and adrenal sequences were identical; the human cDNA and amino acid sequences are 82% and 72% homologous, respectively, with the bovine sequences. P450scc cDNA was used to probe DNA from a panel of mouse-human somatic cell hybrids, showing that the single human P450scc gene lies on chromosome 15. The human P450scc gene is expressed in the placenta in early and midgestation; primary cultures of placental tissue indicate P450scc mRNA accumulates in response to cyclic AMP. PMID- 3024158 TI - Varicella zoster virus glycoprotein gpI is selectively phosphorylated by a virus induced protein kinase. AB - Varicella zoster virus glycoprotein I (VZV gpI; Mr 98,000) was phosphorylated in virus-infected human cell monolayers, while two other major VZV glycoproteins (gpII and gpIII) were not similarly modified. Phosphorylation of VZV gpI was not blocked by inhibitors of glycosylation, nor were the phosphoryl groups enzymatically removed by endoglycosidases. Phosphoamino acid analysis revealed the presence of phosphoserine and phosphothreonine residues on the polypeptide backbone. The selective nature of the phosphorylation event was further demonstrated in vitro by a protein kinase (Mr 50,000), which was present in virus infected cells but absent from uninfected cells or purified virions. The enzyme catalyzed the transfer of 32Pi from [gamma-32P]ATP to gpI but not to gpII and gpIII. Like VZV gpI, this virus-induced protein kinase was also a constituent of the plasma membrane of live VZV-infected cells. PMID- 3024159 TI - Molecular dynamics simulations of cooling in laser-excited heme proteins. AB - In transient optical experiments the absorbed photon raises the vibrational temperature of the chromophore. In heme proteins at room temperature conversion of a 530-nm photon into vibrational energy is estimated to raise the temperature of the heme by 500-700 K. Cooling of the heme is expected to occur mainly by interacting with the surrounding protein. We report molecular dynamics simulations for myoglobin and cytochrome c in vacuo that predict that this cooling occurs on the ps time scale. The decay of the vibrational temperature is nonexponential with about 50% loss occurring in 1-4 ps and with the remainder in 20-40 ps. These results predict the presence of nonequilibrium vibrational populations that would introduce ambiguity into the interpretation of transient ps absorption and Raman spectra and influence the kinetics of sub-ns geminate recombination. PMID- 3024160 TI - Temporal comparisons in bacterial chemotaxis. AB - Responses of tethered cells of Escherichia coli to impulse, step, exponential ramp or exponentiated sine-wave stimuli are internally consistent, provided that allowance is made for the nonlinear effect of thresholds. This result confirms that wild-type cells exposed to stimuli in the physiological range make short term temporal comparisons extending 4 sec into the past: the past second is given a positive weighting, the previous 3 sec are given a negative weighting, and the cells respond to the difference. cheRcheB mutants (defective in methylation and demethylation) weight the past second in a manner similar to the wild type, but they do not make short-term temporal comparisons. When exposed to small steps delivered iontophoretically, they fail to adapt over periods of up to 12 sec; when exposed to longer steps in a flow cell, they partially adapt, but with a decay time of greater than 30 sec. cheZ mutants use a weighting that extends at least 40 sec into the past. The gain of the chemotactic system is large: the change in occupancy of one receptor molecule produces a significant response. PMID- 3024161 TI - Localization of Epstein-Barr virus-encoded small RNAs by in situ hybridization. AB - Human B lymphocytes latently infected with Epstein-Barr virus (EBV) synthesize two low molecular weight RNAs designated EBER 1 and 2. Using an in situ hybridization technique we have localized EBER 1 and 2 within the nucleus of single EBV-harboring B lymphocytes from established and recently transformed cell lines. As controls, the locations of the small nuclear RNA, U1, and the small cytoplasmic RNA, 7SL, were examined in HeLa and EBV-harboring cells. Because of possible functional similarities between EBERs and the adenovirus-associated (VA) RNAs, VAI was also localized; it appeared to be in the nucleus and cytoplasm, implying that VAI may have a different role than that of the nuclear-localized EBERs. PMID- 3024162 TI - Artificial mitochondrial presequences. AB - Synthetic oligonucleotides were used to construct artificial mitochondrial presequences that contained, besides the initiator methionine, only arginine, serine, and leucine. The ratio of these three amino acids was adjusted to match that of basic, hydroxylated, and hydrophobic residues in natural mitochondrial presequences. When these sequences were fused to the N terminus of yeast cytochrome oxidase subunit IV lacking its own presequence, they directed the attached subunit IV to its correct intramitochondrial location in vivo. They also mediated import of subunit IV into isolated yeast mitochondria. In contrast, artificial sequences containing glutamine, arginine, and serine residues following the initiator methionine were inactive. Thus, the targeting function of mitochondrial presequences does not depend on specific amino acid sequences but may instead depend on the overall balance between basic, hydrophobic, and hydroxylated amino acids. PMID- 3024163 TI - Membrane-associated inhibitor of DNA synthesis in senescent human diploid fibroblasts: characterization and comparison to quiescent cell inhibitor. AB - Cell membranes prepared from senescent human diploid fibroblasts (HDF) inhibited entry into S phase by 35% when added to the medium of replicating young HDF. This membrane-associated inhibitory activity was (i) sensitive to trypsin, heat, and periodate, which suggests that the inhibitor is a glycoprotein, and (ii) not able to inhibit DNA synthesis in simian virus 40-transformed HDF, which indicates that not all types of cells are sensitive to this inhibitor. Quiescent young HDF also have a surface membrane-associated inhibitor of DNA synthesis. A comparison of the senescent HDF and quiescent HDF inhibitor activities indicates that they may have the same chemical and physical nature and the same specific activity, but their regulation is different. The inhibitory activity of quiescent young HDF is abolished within 20 hr after refeeding with fresh serum-containing medium, whereas that of senescent HDF remains unchanged. Quiescent old HDF (two or three population doublings remaining) exhibit an intermediate response to serum with approximately two-thirds of the inhibitory activity abolished. The fraction of cells in S phase at 20-24 hr post-stimulation (37% in young HDF, 24% in old HDF, and 0% in senescent HDF) is inversely proportional to inhibitor levels. This suggests that inability to neutralize the inhibitory activity in response to serum stimulation could be involved in the inability of senescent HDF to enter S phase. Disappearance of the inhibitory activity from quiescent young HDF occurs late in G1 phase. Thus, the inhibitor may play a role in determining the length of the G0 to S phase transition in these cells. PMID- 3024164 TI - Transgenesis by means of blastocyst-derived embryonic stem cell lines. AB - This study demonstrates that blastocyst-derived embryonic stem cells (ES cells) can be used as a vehicle for transgenesis. The method is nearly as efficient as other methods, and the introduced neomycin phosphotransferase (neo) gene is stably transmitted through several generations with no apparent loss in G418 resistance. An important factor contributing to the efficiency of this process is the rigorous selection, before blastocyst injection, of genetically transformed cells for in vitro developmental pluripotency. One of the advantages of the ES cell route to transgenesis is that it provides investigators with the opportunity to screen for the desired genetic alterations before reintroducing the ES cells into the animal. PMID- 3024165 TI - Escherichia coli K-12 restricts DNA containing 5-methylcytosine. AB - We have observed that plasmids containing certain cloned modification methylase genes of type II restriction-modification systems cannot be transformed into many laboratory strains of Escherichia coli K-12. The investigation of this phenomenon, reported here, has revealed (i) DNA containing 5-methylcytosine is biologically restricted by these strains, while DNA containing 6-methyladenine is not; (ii) restriction is due to two genetically distinct systems that differ in their sequence specificities, which we have named mcrA and mcrB (for modified cytosine restriction). Since 5-methylcytosine containing DNA is widespread in nature, the Mcr systems probably have a broad biological role. Mcr restriction may seriously interfere with molecular cloning of 5-methylcytosine-containing foreign DNAs. The Mcr phenotypes of some commonly used strains of E. coli K-12 are reported. PMID- 3024166 TI - A method for identifying the viral genes required for herpesvirus DNA replication. AB - Several laboratories have shown that transfected plasmid DNAs containing either of the two known origins of herpes simplex virus (HSV) DNA replication, oriS or oriL, are replicated in HSV-1-infected cells or in cells cotransfected with virion DNA. I have found that HSV-1 (KOS) DNA digested to completion with the restriction enzyme Xba I is as efficient as intact viral DNA in supporting the in vivo replication of cotransfected plasmids containing oriS. On the basis of this result, several of the Xba I restriction fragments of HSV-1 DNA were cloned into the plasmid vector pUC19, and combinations of cloned DNAs were tested for their ability to supply the trans-acting functions required for HSV origin-dependent replication. A combination of five cloned fragments of HSV-1 can supply all of the necessary functions: Xba I C (coordinates 0.074-0.294), Xba I F (coordinates 0.294-0.453), Xba I E (coordinates 0.453-0.641), Xba I D (coordinates 0.641 0.830), and EcoRI JK (coordinates 0.0-0.086; 0.830-0.865). Transient plasmid replication in this system is dependent on the presence of either oriS or oriL in cis. The plasmid containing Xba I F can be replaced by two smaller plasmids, one of which contains only the gene for the HSV-encoded DNA polymerase, and the other of which contains only the gene for the major DNA binding protein (ICP8). Thus, plasmid DNA replication in this system depends on two of the genes known from genetic studies to be essential for viral DNA replication in infected cells. This system defines a simple complementation assay for cloned fragments of HSV DNA that contain other genes involved in viral DNA replication and should lead to the rapid identification of all such genes. PMID- 3024167 TI - Stimulatory and inhibitory influences of human immunodeficiency virus on normal B lymphocytes. AB - B-lymphocyte dysfunction is a characteristic feature of the acquired immunodeficiency syndrome (AIDS) and of the AIDS-related complex. The aim of the present study was to further examine the influences exercised by the human immunodeficiency virus (HIV; formerly called human T-lymphotropic virus type III or lymphadenopathy-associated virus, HTLV-III/LAV) on normal human B lymphocytes. An unfractionated protein preparation, made from HIV purified by density gradient centrifugation, was previously shown to induce differentiation of normal human B lymphocytes into immunoglobulin-secreting cells. In the present analyses, this B lymphocyte response peaked on day 6 or 7 after culture initiation and was found to be independent of the requirement for monocytes but to require T cells. Responses could also be elicited in cultures of purified B cells by the addition of T cells that had been exposed to HIV antigen. Inhibitors of protein synthesis (puromycin and cycloheximide) abrogated the responses. In contrast to its stimulatory effects, the same virus preparation was previously shown to inhibit polyclonal responses that are normally elicited in peripheral blood lymphocyte cultures by a T-dependent stimulus (pokeweed mitogen) and T-independent stimulus (Epstein-Barr virus). The present studies suggest that the inhibitory effects of the HIV antigen studied herein are targeted primarily at the B lymphocytes. The role of T lymphocytes in the HIV antigen-mediated inhibitory effects, although demonstrated, could not be conclusively established as an essential pathway. These findings elucidate mechanisms by which components of HIV exert stimulatory as well as inhibitory effects on human B lymphocytes and thereby lead to the dysfunction of these cells in HIV infection. PMID- 3024168 TI - Mechanism of action of some inhibitors of endothelium-derived relaxing factor. AB - The mechanism of the inhibitory action of phenidone, 3-amino-1-[m (trifluoromethyl)phenyl]-2-pyrazoline (BW 755C), dithiothreitol, hydroquinone, and pyrogallol on the vascular relaxation induced by endothelium-derived relaxing factor (EDRF) was investigated. EDRF was released from porcine aortic endothelial cells in culture and bioassayed on a cascade of superfused rabbit aortic strips. These compounds inhibited EDRF-induced relaxation of vascular strips, without affecting the relaxation induced by glyceryl trinitrate, and their inhibitory potency was markedly attenuated (by more than 1 order of magnitude) by the addition of superoxide dismutase (5-15 units/ml) or oxidized cytochrome c (20-40 microM) but not by catalase (30 units/ml) or heat-inactivated superoxide dismutase. These data indicate that the above five inhibitors inactivate EDRF through the formation of superoxide ions, which have recently been shown to destroy EDRF. The inhibition of EDRF by these compounds is therefore attributable to their redox properties rather than to any specific biological action. PMID- 3024169 TI - Sequence homology between acquired immunodeficiency syndrome virus envelope protein and interleukin 2. AB - A region of homology has been found between the envelope (env) protein of the acquired immunodeficiency syndrome (AIDS) virus and a portion of interleukin 2 (IL-2) that purportedly binds to the IL-2 receptor. This homology, between two proteins associated with opposing biological functions, suggests possible mechanisms for the immunosuppressive activity of the AIDS virus. Two mechanisms are proposed in which the AIDS virus env protein interferes with IL-2 activity either directly or indirectly. A region of similarity to the purported IL-2 receptor binding site on IL-2 and AIDS virus env is present in the env proteins of other retroviruses associated with immunosuppression. A synthetic peptide vaccine for AIDS is suggested based on the IL-2 receptor binding sequence in AIDS virus env. PMID- 3024170 TI - Activation of phospholipases A and C in human platelets exposed to epinephrine: role of glycoproteins IIb/IIIa and dual role of epinephrine. AB - Human platelets stimulated by epinephrine undergo enhanced turnover of phosphatidylinositol 4,5-bisphosphate, accumulate inositol trisphosphate, diacylglycerol, and phosphatidic acid, and phosphorylate a 47-kDa protein. All of these phenomena indicate stimulation of phospholipase C. These responses are blocked completely by inhibitors of alpha 2-adrenergic receptors (yohimbine), cyclooxygenase (aspirin or indomethacin), phospholipase A [2-(p amylcinnamoyl)amino-4-chlorobenzoic acid (ONO-RS-082)], Na+/H+ exchange [ethylisopropylamiloride (EIPA)], fibrinogen binding to glycoprotein IIb/IIIa (antibody A2A9), Ca2+/Mg+ binding (EDTA), or removal of fibrinogen. Epinephrine evokes (i) an increased turnover of ester-linked arachidonic acid in aspirin treated platelets that is inhibited by ONO-RS-082, EDTA, yohimbine, or the absence of fibrinogen and (ii) a rapid cytoplasmic alkalinization that is inhibited partially by blockage of cyclooxygenase activity and completely by A2A9 or EIPA. In contrast, when incubated with subaggregatory concentrations of the prostaglandin H2/thromboxane A2 analogue [(15S)-hydroxy-11 alpha,9 alpha (epoxymethano)prosta-5,13-dienoic acid (U46619) and epinephrine, aspirin-treated platelets show a potentiation of phospholipase C activation that is unaffected by the above inhibitors. We propose that epinephrine, in promoting exposure of glycoprotein IIb/IIIa sites for fibrinogen binding, leads to a cytoplasmic alkalinization, which, in conjunction with local shifts in Ca2+, promotes low level activation of phospholipase A. The resulting free arachidonic acid is converted to cyclooxygenase products, which, potentiated by epinephrine, activate phospholipase C. This further amplifies the initial stimulatory response. PMID- 3024171 TI - Nondefective spleen necrosis virus-derived vectors define the upper size limit for packaging reticuloendotheliosis viruses. AB - We constructed a nondefective retrovirus vector based on spleen necrosis virus (SNV), a replication-competent reticuloendotheliosis virus. We introduced different DNA sequences into this vector and studied the ability of the resulting viruses to replicate in chicken embryo fibroblasts. The replication efficiency of SNV-derived viruses decreased with increasing virus size. Viruses larger than 9.4 kilobases (kb) were rapidly overgrown by replication-competent deletion mutants. The size restriction for the efficient replication of nondefective SNV-derived viruses prevented the production of viruses larger than 10.0 kb. Analysis of the kinetics of virus particle release indicated that the size restriction occurred during virus encapsidation. PMID- 3024172 TI - Purification of a benzodiazepine from bovine brain and detection of benzodiazepine-like immunoreactivity in human brain. AB - An endogenous brain substance that binds to the central-type benzodiazepine receptors with agonist properties is present in both rat and bovine brains. This substance has been purified to homogeneity from bovine brain by immunoaffinity chromatography on immobilized monoclonal anti-benzodiazepine antibody followed by gel filtration on Sephadex G-25 and two reversed-phase HPLC steps. The purified substance was characterized as the benzodiazepine N-desmethyldiazepam (nordiazepam). The techniques used for the identification were mass spectrometry, HPLC, spectrophotometry, benzodiazepine receptor binding, and immunological techniques. Benzodiazepine-like immunoreactivity was also found in all the human brains tested, including six brains that had been stored in paraffin since 1940, fifteen years before the first synthesis of benzodiazepines. These results show that benzodiazepine-like molecules of natural origin--and possibly benzodiazepines themselves--are present in human and other mammalian brains. PMID- 3024173 TI - Phosphoinositide hydrolysis and smooth muscle function. PMID- 3024174 TI - Fat, lipotropes, hypolipidemic agents and liver cancer. PMID- 3024175 TI - Fat, calories and fiber. PMID- 3024176 TI - The epidemiology of colon cancer and dietary fat. PMID- 3024177 TI - Pharmacology and experimental basis of therapy with LHRH agonists in women. PMID- 3024178 TI - Extra pituitary actions of LHRH analogues in tissues of the human female and investigation of the existence and function of LHRH-like peptides. PMID- 3024179 TI - Increase in beta- and alpha 1-adrenoceptor binding sites in the rat brain and in the alpha 1-adrenoceptor functional sensitivity after the DSP-4-induced noradrenergic denervation. AB - Changes in the density of beta- and alpha 1-adrenoceptors were studied following denervation of the rat cerebral cortex and hippocampus, caused by systemic administration of DSP-4. The noradrenergic denervation increased both beta- and alpha 1-adrenoceptor density by about 30 and 17%, respectively in the cortex, and by about 30% in the hippocampus. In order to estimate the behavioral response of normal and DSP-4-treated rats to alpha 1-agonist, the influence of phenylephrine (25 micrograms ICV) on the exploratory activity of rats in the open field was measured. Phenylephrine failed to change the exploratory activity of normal rats, but significantly increased it in DSP-4 animals. The results indicate that noradrenergic denervation produces an increase in number of both beta- and alpha 1-adrenoceptors and the functional supersensitivity to the alpha 1-adrenergic agonists. PMID- 3024180 TI - Effect of (1-24)adrenocorticotrophin stimulation on the rate of corticosterone synthesis by the rat adrenal cortex. AB - The rate of corticosterone synthesis by the rat adrenal gland was measured in vitro, using a cell-free system, following the in vivo administration of (1 24)ACTH. The doses of ACTH used were 50, 100 and 250 micrograms ACTH/kg body weight. With all 3 doses of ACTH there was a significant increase in the rate of corticosterone synthesis within the first 5 min; this initial increase in rate did not vary with the dose of ACTH given. With both the 50 and 100 micrograms/kg doses the new rate of synthesis was maintained without further change, up to 30 min post-injection. In the case of the 250 micrograms/kg dose there was a second significant increase in the rate of corticosterone synthesis observed after 20 min. The results are discussed in the light of the hypothesis that the differential response of the adrenal gland represents the binding of ACTH to two receptors; a high affinity receptor of low abundance and a low affinity receptor of greater abundance. The results are consistent with the initial steroidogenic response resulting from binding to the high affinity receptors. Because of their low abundance these receptors may be completely occupied even at low doses of ACTH, thus explaining the dose-independent nature of the initial response to ACTH. Binding to the more abundant low affinity receptors may be associated with the secondary dose-dependent enhanced synthesis rate, a response which may be mediated via c-AMP. PMID- 3024181 TI - Metabolic interactions of phencyclidine (PCP) and delta 9-tetrahydrocannabinol (THC) in the rat. AB - The in vitro effects of THC on the metabolism of PCP by rat liver were determined. Samples containing 1 mM PCP were incubated for 1 hr at 37 degrees C with an NADPH-generating system containing 10,000 X g supernatant or Ca++ precipitated rat liver microsomes. These incubations were carried out in the presence or absence of THC and at the end of 1 hr, PCP metabolites were determined by gas chromatography. In the presence of 0.1, 0.05, 0.025 and 0.0125 mM THC, the production of 1-(1-phenyl-4-hydroxycyclohexyl)piperidine (metabolite I) by the 10,000 X g supernatant was decreased by 46, 29, 23 and 16% respectively. Similarly, production of 1-(1-phenylcyclohexyl)-4-hydroxypiperidine (metabolite II) was reduced significantly by 58, 44, 34 and 23% with the respective concentrations of THC. However, the production of 1 phenylcyclohexylamine (metabolite III) was increased by 18, 32, 30 and 22% with 0.1, 0.05, 0.025 and 0.0125 mM THC. Incubations with Ca++-precipitated liver microsomes revealed similar trends in PCP metabolism in the presence or absence of THC. Metabolites I and II were reduced by 62 and 67% by 0.1 mM THC. Another concentration of THC (0.025 mM) caused a 50 and 62% decrease in I and II. These observations suggest that THC alters the in vitro microsomal metabolism of PCP. PMID- 3024182 TI - Alpha-adrenoceptors and monoamine contents in the cerebral cortex of the rodent Jaculus orientalis: effects of acute cold exposure. AB - The tritiated adrenergic antagonists prazosin ([3H]PRZ) and idazoxan ([3H]IDA, or RX-781094) bind specifically and with high affinity to alpha 1- and alpha 2 adrenoceptors respectively, and were used to measure adrenoceptors in membrane preparations obtained from the cerebral cortex of Jaculus orientalis. Membrane preparations were also obtained from a group of cold exposed animals, to determine whether these adrenoceptors could be modified by a thermic stress. The density of receptors (Bmax; maximum binding capacity) and the dissociation constant (Kd 25 degrees C) were estimated by iterative modelling, and by using the procedure of Hill. After acute cold exposure (16 hr, 5 degrees C) there was a decrease in the affinity of the alpha 1-adrenoceptors, as judged by the Kd 25 degrees C for [3H]PRZ, with no changes in the Bmax. The alpha 2-sites did not show any significant changes, as revealed by [3H]IDA binding. Pretreatment of the membrane preparations from control animals with the disulfide and sulfhydryl reactives DL-dithiothreitol, 5,5'-dithiobis-(2-nitrobenzoic acid) and N ethylmaleimide decreased specific [3H]PRZ and [3H]IDA binding, with minor changes in non-specific counts, indicating that the fixation of these ligands was to the receptor proteins. The endogenous cortical monoamine contents were also determined in the frontal cerebral cortex of these same animals, using high performance liquid chromatography with electrochemical detection. The catecholamine levels and their major metabolites were found to be stable in the cortex after the acute thermic stress, but there was a marked reduction in serotonin with a normal content in 5-hydroxyindole-3-acetic acid. PMID- 3024183 TI - Discriminative stimulus properties of phencyclidine (PCP)-related compounds: correlations with 3H-PCP binding potency measured autoradiographically. AB - Several PCP analogs, the putative PCP agonist MDP, and the sigma receptor agonists SKF-10,047 and dexoxadrol were tested for their ability to substitute for PCP in animals trained to discriminate PCP from saline. The potencies of these compounds in substituting for PCP in the behavioral task correlated with their abilities to inhibit the specific binding of 3H-PCP to rat hippocampal sections measured autoradiographically, which occurred at a single class of sites with an affinity of 85 nM and a capacity of 2646 fmol/mg protein. In addition to this specific binding, an additional nonspecific but displaceable fraction of total 3H-PCP binding was present. These results suggest that the specific 3H-PCP binding site measured in the hippocampus may be the type of binding site which mediates the behavioral effects of PCP and related compounds. Therefore, measurement of the inhibition of 3H-PCP binding at this site might aid in the search for PCP antagonists. PMID- 3024184 TI - Comparison of anticonvulsant effect of pentobarbital and phenobarbital against seizures induced by maximal electroshock and picrotoxin in rats. AB - Pentobarbital and phenobarbital exhibited anticonvulsant effect against maximal electroshock (MES) and picrotoxin-induced seizures in rats. Bicuculline, a GABAA receptor antagonist, reversed the anticonvulsant effect of pentobarbital, but not of phenobarbital, at a dose having no effect per se. Although picrotoxin (2 mg/kg, IP) potentiated MES seizures, it did not reverse the anticonvulsant effect due to either pentobarbital or phenobarbital. GABAB receptor antagonists such as delta-amino-n-valeric acid and homotaurine failed to modify the anticonvulsant effect due to pentobarbital or phenobarbital. Furthermore, GABAA agonist muscimol but not baclofen, a GABAB receptor agonist, exhibited the anticonvulsant effect against MES-induced seizures. However, baclofen when combined with sub-effective dose of pentobarbital or phenobarbital offered protection against MES seizures. Pentobarbital and phenobarbital were effective in almost equivalent doses against MES, as well as against picrotoxin-induced seizures. These observations indicated that pentobarbital exhibits anticonvulsant effect against MES seizures through the involvement of GABAA receptors, and activation of GABAB receptors alone does not seem to play any significant role in MES seizures and in the anticonvulsant effect of pentobarbital. However, activation of GABAB receptor does potentiate the facilitatory effect of barbiturates on GABAAergic transmission and in their anti-MES effect. Moreover, these results also suggest that the anticonvulsant effect of barbiturates against MES-seizures may involve other mechanisms in addition to GABAAergic transmission. PMID- 3024185 TI - Brain asparaginase, ACE activity and plasma cortisol level in morphine dependent rats: effect of aspartic acid and naloxone. AB - The activities of the brain L-asparaginase and angiotensin converting enzyme (ACE), and the plasma cortisol level were found to be decreased in the rats implanted with morphine (M) containing pellets. Even though 10 mg/kg of naloxone (N) itself showed an inhibitory effect on ACE it abolished the inhibitions seen in the M dependent rats five min following subcutaneous injection. The chronic administration of L-aspartic acid (ASP) during the development of physical dependence or just before the N injection prevented the increase of the plasma cortisol caused by N. It is concluded that in addition to the inhibition of the brain L-asparaginase activity which was previously hypothesized to be the main reason of the development of physical dependence on opiates as a result of the related experimental and clinical data, the inhibition by M of the brain ACE activity may take part in the development of physical dependence. With regard to the plasma cortisol level, the concomitant administration of ASP with M blocks, to a great extent, the development of physical dependence on opiate. The single dose of ASP administration before N injection prevents the effect of N, the manifestation of abstinence syndrome. PMID- 3024186 TI - Suppression of behavior by food pellet-lithium chloride pairings in squirrel monkeys. AB - Responding by squirrel monkeys was maintained under a 30-response fixed-ratio schedule of food presentation; during different sessions responding produced either sucrose-flavored or banana-flavored food pellets. Pre-session administration of doses of lithium chloride (LiCl) less than 3.0 mEq/kg did not alter rates of responding whereas pairing either type of pellet with post-session injections of 1.8 mEq/kg LiCl suppressed both lever pressing and consumption during subsequent sessions in which that pellet type was available. When post session injections of LiCl were discontinued, responding recovered within 14 sessions. The suppression of responding, but not pellet consumption, was then reliably reproduced in each monkey by pairing post-session LiCl with the previously non-paired type of pellet. Pre-session administration of chlordiazepoxide (CDAP, 3.0-17.0 mg/kg) increased rates of suppressed responding in a dose-related manner, but did not increase pellet consumption. These data indicate that different mechanisms may be involved in the suppression of responding and the suppression of consumption of food by post-session injections of drugs. The suppression of responding by post-session injections of drugs in primates appears to be qualitatively similar to the suppression of responding by other noxious stimuli such as electric shock in that it is reversible, it can be reinstated by re-exposure to post-session drug injections, and it can be attenuated by pre-session administration of CDAP. PMID- 3024187 TI - A measurement of w for 150 MeV protons in nitrogen and argon. AB - Ionisation chamber dosimetry for proton radiation therapy requires accurate measurements of w, the average energy necessary to produce an ion pair for protons in the therapeutic energy range of 30-240 MeV. In this investigation, such measurements were made at the Harvard Cyclotron Laboratory for 150 MeV protons in nitrogen and argon with an accuracy of 2%. Ionisation and energy loss were measured simultaneously in a gas cell 1 m in length. A parallel plate ionisation chamber situated inside the gas cell was used to measure ionisation, and the energy loss was determined from time-of-flight and range measurements. The data indicate that w is 26.5 +/- 0.6 eV/(ion pair) for argon and 36.3 +/- 0.8 eV/(ion pair) for nitrogen. These results are consistent with measurements at lower energies. PMID- 3024188 TI - Rheumatoid arthritis of the shoulder. AB - The shoulder frequently is affected by rheumatoid arthritis (RA), often resulting in musculoskeletal dysfunction. The primary purpose of this article is to provide the clinician with an in-depth look at the pathological factors and treatment of RA as it affects the shoulder. The review includes sections on pathological factors, radiology, morbidity, clinical manifestations, and both conservative and surgical management. The secondary purpose of this article is to acquaint the clinician with some of the other rheumatic diseases in which shoulder dysfunction may occur. I hope that the presentation of management principles will stimulate discussion and interest regarding the scientific rationale for the treatment of musculoskeletal manifestations of RA. PMID- 3024189 TI - Antiviral activity of the photoactive thiophene alpha-terthienyl. PMID- 3024190 TI - Further studies on the antiviral activity of harmine, a photoactive beta carboline alkaloid. PMID- 3024191 TI - Effects of learned flavor avoidance on grooming behavior in rats. AB - In Experiment 1, rats were conditioned to avoid saccharin in tapwater by pairing it with LiCl in carboxymethylcellulose (CMC) applied to the fur. Conditioned flavor avoidance (CFA) of saccharin was then assessed in drinking and grooming tests. In Experiment 2, rats were given saccharin CMC on their fur and NaCl in water (or vice-versa) as conditioned stimuli in a CFA paradigm. Two-choice tests (saccharin vs. NaCl) followed in drinking and grooming contexts. In Experiment 3, rats were given saccharin CMC on one flank and vehicle (CMC only) on the other. After grooming, animals were injected with LiCl and then given 2-choice tests, first between saccharin and water, then between saccharin-CMC and plain-CMC, and finally, between saccharin and water. Strong CFA was exhibited in drinking tests in all 3 experiments. This was not the case in grooming tests. Rats continued to groom when tastant was applied to only one flank (Experiment 1), and exhibited only weak CFA when a different tastant was applied to each flank (Experiments 2 and 3). We conclude that grooming can be directed to minimize the ingestion of noxious substances, but that such ingestion is not sufficiently reduced to affect the efficacy of grooming as a delivery method for unpalatable substances (e.g., rodenticides, chemosterilants). We speculate that grooming represents a weakness in rodents' defenses against dietary poisoning, and that it might be used to deliver toxicants as part of crop protection schemes that make use of CFA. PMID- 3024192 TI - Effects of estrous cycles and ovarian steroids on body weight and energy expenditure in Syrian hamsters. AB - In Experiment 1, highly significant changes were observed over the estrous cycle in body weight gain, but not in food intake, daytime resting oxygen consumption or brown fat thermogenesis in Syrian hamsters. In Experiments 2 and 3, body weight and composition, food intake, resting oxygen consumption, and brown fat thermogenesis were measured following estradiol or estradiol plus progesterone treatment in ovariectomized hamsters. The significant changes in body weight could not be explained by changes in food intake, and were not accompanied by significant alterations in daytime oxygen consumption or brown fat thermogenic activity. In Experiment 4, resting oxygen consumption and body weight were measured every 6 hours over the estrous cycle. There was a striking absence of the usual nocturnal peak in resting oxygen consumption on the night of estrus (the night of the largest body weight gain). However, brown fat thermogenic activity did not differ among groups of hamsters killed on different nights of the estrous cycle. Estradiol-induced changes in energy storage may be mediated by changes in the daily rhythm of energy expenditure which are not dependent on alterations in brown fat thermogenesis. PMID- 3024193 TI - Age-dependent day/night variations of alpha 1- and beta-adrenoceptors in the rat cerebral cortex. AB - Receptor binding studies in the cerebral cortex of young (2-4 months) and mature (11-12 months) rats were conducted at two points of the circadian time: 8.00 and 20.00 hr. In addition, the exploratory activity in the open field was assessed. In young rats the density of alpha 1- and beta-adrenoceptors increased at 20.00 in comparison with 8.00 hr, while no changes in the exploratory activity were observed. However, in mature rats the activity was higher at 20.00 than 8.00 hr; at the same time, a rise in the density of alpha 1- but not beta-adrenoceptors was observed. Obtained data suggest that the increased exploratory activity of mature rats at 20.00 hr may appear as a result of a higher density of alpha 1 adrenoceptors and a simultaneous lack of changes in beta-adrenoceptors. PMID- 3024194 TI - Disruption of conditioned taste aversion: the effect of ECS after the taste illness interval. AB - Rats were taught a conditioned taste aversion (CTA) by pairing a 10% sucrose solution (CS) with lithium chloride-induced poisoning (UCS) 30 min later. The extent to which electroconvulsive shock (ECS) (80 mA for 600 msec) impaired the acquisition of the CTA was studied by either interpolating the ECS within the CS UCS interval or by administering it at various times following the UCS (0, 5, 10, 15, 30, 60 and 120 min). There was a pronounced although limited loss of learning among all groups when ECS was delivered no later than 10 min after the UCS. When ECS was administered between 15 and 120 min, the overall aversion acquired did not differ significantly from that of the poisoned controls. It is concluded that while the stability of CTA seems to increase at about the same time the behavioral concomitants of lithium toxicity become apparent (10 min), this phenomenon does not necessarily imply that the neural trace underlying CTA has consolidated into a less labile state. PMID- 3024195 TI - Oxytocin release during coitus in male and female rabbits: effect of opiate receptor blockade with naloxone. AB - Blood was collected from freely moving rabbits pretreated with saline or naloxone both before and after coitus. In the males, ejaculation was associated with a marked increase in plasma oxytocin concentrations. In the females, oxytocin release was an all-or-nothing phenomenon-either there was no change in oxytocin or it rose to levels very similar to that found post-ejaculation in the males. Naloxone did not alter the proportion of occasions in which females showed an oxytocin response, nor did it have a significant effect on the levels of oxytocin achieved during coitus in either the male or the female rabbits. PMID- 3024196 TI - Tetrahydrocannabinol content and differences in marijuana smoking behavior. AB - Changes in smoking behavior in response to a change in marijuana potency were measured in marijuana users. A marijuana cigarette containing 1.2% or 3.9% tetrahydrocannabinol (THC) was smoked on separate days by ten experienced users. Puff volume, duration and number, interpuff interval, inhalation volume and duration were averaged for each cigarette. The high potency cigarettes were smoked with more puffs and longer interpuff intervals, but also with greater inhaled volumes of air, thereby diluting the marijuana smoke. PMID- 3024197 TI - An observational analysis of the effect of the selective kappa opioid agonist, U 50,488H, on feeding and related behaviours in the rat. AB - The behaviour of partially pre-satiated rats consuming a sweet palatable food and treated with either vehicle or the specific kappa receptor agonist U-50,488H (0.1 3 mg/kg) was recorded on videotape. Analysis revealed that the hyperphagia induced by the kappa agonist (0.3-3 mg/kg) resulted from an increase in the duration of feeding and not from an increase in the local rate of eating. The increase in duration was due, in turn, to a greater frequency of bouts of feeding. The kappa agonist also increased the latency to the final feeding bout. The effect of U-50,488H was consistent with de-satiation, so that the increase in feeding duration was in evidence from the start of the test period, while the temporal pattern of later satiation was preserved but lagged behind that of control animals. At the largest dose, other recorded activities (rearing, locomotor activity, grooming) were suppressed, with a marked increased in inactivity. At the lowest dose (0.1 mg/kg) there was a significant increase in grooming behaviour. The results are discussed with reference to an hypothesis of opioid function in the control of food intake. PMID- 3024198 TI - The possible modulation of morphine analgesia by the supramolecular GABA receptor complex. AB - In the present study, the mechanism of the antagonistic action of 0.5 mg/kg diazepam on the analgesic effect of morphine was investigated. While Ro 15-1788, a benzodiazepine receptor antagonist, was found to partially reverse the inhibitory action of diazepam on morphine analgesia, a chloride channel blocking agent, picrotoxin, produced complete antagonism of the action of diazepam. Furthermore, picrotoxin potentiated the partial antagonistic effect of Ro 15-1788 at a normally ineffective dose to affect the 0.5 mg/kg diazepam-morphine dose response curve. These overall effects of picrotoxin on the supramolecular GABA receptor complex are discussed. PMID- 3024199 TI - Increased central alpha 2-adrenoceptor sensitivity in panic disorder. AB - Cardiovascular responses to an intravenous challenge dose of clonidine (1.5 micrograms/kg) were measured in eight patients with DSM III panic disorder. In comparison with an age- and sex-matched control population panic patients showed significantly greater falls in systolic and diastolic blood pressure, with similar falls in heart rate. These observations support the view of a biological abnormality in panic disorder. PMID- 3024200 TI - Morphological, biochemical and secretory studies on rat pancreatic ducts maintained in tissue culture. AB - Interlobular ducts were isolated from the pancrease of copper-deficient rats and maintained in culture on polycarbonate filter rafts. Within 8 h the ends of the ducts had sealed. This was followed by a marked dilatation of the lumen, a flattening of the epithelium against the surrounding connective tissue layer and an over-all swelling of the duct. Apart from a reduction in their height, a fall in the number of intracellular fat droplets and a widening of intercellular spaces, epithelial cells within the cultured ducts retained all the ultrastructural characteristics of those in freshly isolated preparations. The basal concentration of adenosine 3',5'-phosphate (cyclic AMP) in the cultured ducts was 43.3 +/- 6.8 mumol.1-1 duct epithelium (n = 5) and was increased to 188.9 +/- 55.2 mumol.l-1 duct epithelium (n = 5) in the presence of 10(-8) M secretin. The basal rate of fluid secretion, measured using micropuncture techniques, was 0.16 +/- 0.03 nl.h-1.nl-1 duct epithelium (n = 12). This was increased 14-fold by 10(-8) M secretin while the dose of the hormone required for half-maximal secretion was about 2 X 10(-11) mol.l-1. The concentration of chloride ions in secreted fluid and perifusion buffer were similar. Variation in culture time up to 52 h had no effect on fluid secretion, and the response to secretin was dependent on the presence of bicarbonate ions in the perifusion fluid. N6,O2-dibutyryl adenosine 3',5'-phosphate (dibutyryl cyclic AMP) also increased fluid secretion but caerulein had no effect. We suggest that isolated ducts secrete fluid at comparable rates to ducts in situ within the pancrease of copper-replete rats. PMID- 3024201 TI - Reactions of electron-transfer proteins at electrodes. PMID- 3024203 TI - Induction of L-arabinose isomerase in gamma-irradiated Escherichia coli. AB - Gamma irradiation of Escherichia coli B/r caused a dose-dependent inhibition of the capacity of the cells to synthesize L-arabinose isomerase in response to the inducer. At higher doses (18 krad and above), postirradiation incubation led to further inhibition of the capacity to synthesize L-arabinose isomerase, whereas cells receiving lower doses recovered from the damage to the enzyme synthesizing system following incubation. Cyclic AMP partially reversed the inhibitory effect on L-arabinose isomerase induction produced immediately after irradiation by all gamma-ray doses (up to 30 krad), but the enhanced inhibitory effect caused by induction in cells irradiated at higher doses could not be reversed by the nucleotide. It is suggested that although catabolite repression is partly responsible for causing the inhibition of the enzyme synthesizing capacity of the cells observed immediately after gamma irradiation, the enhanced inhibition caused by incubating cells irradiated at higher doses is not due to interference with the control mechanism regulated by catabolite repression. PMID- 3024202 TI - Comparison of the protective effects of three phosphorothioate radioprotectors in the RIF-1 tumor. AB - Three aminoalkyl phosphorothioates, WR-2721, WR-3689, and WR-77913, were compared as radioprotectors of RIF-1 tumors irradiated in vivo and assayed for cell survival in vitro. The protector doses were 50% of the acute drug LD50. The radiation dose modifying factors for the three drugs were nearly equal, ranging from 1.5 to 1.7 at surviving fractions of 0.1 and 0.05. Using biodistribution data obtained with 35S labeled drugs, the uptake in tumors was calculated as micromoles drug per gram of tumor. On this basis, tumor levels of WR-77913 were 4.5-fold those of WR-2721, and WR-3689 uptake was 2.7-fold greater than uptake of WR-2721. Thus, on a molar basis, WR-2721 appears to be the most effective protector, but all three phosphorothioates protect this tumor moderately well. In diffusible substance autoradiographs of 3H WR-3689 labeled tumors, label was generally distributed over cells with no evidence of preferential localization over nuclei. PMID- 3024204 TI - [Pulmonary microlithiasis following iodine lipiodol lymphography?]. AB - Pulmonary fibrosis with marked calcifications in an 83 year old female patient is reported. Following previously inconspicuous thoracic x-ray findings (1975 and 1978) a bipedal oil lymphography was carried out for diagnosis of a systemic malignant lymphoma. The calcifying pulmonary fibrosis was investigated with the help of tomography, computed tomography, xeroradiography and scintigraphy of the lung and the skeletal system as well as magnetic resonance imaging. A biopsy was refused by the patient. The etiological possibilities of an idiopathic disease as well as an induction by the oil contrast medium and a concomitant malignant lymphoma are discussed. PMID- 3024205 TI - [Roentgen symptoms of early cancer of the breast]. AB - Based on 68 verified cases of early breast cancer, the radiologic signs in early breast cancer are analyzed. The relative incidence of the different mammographic signs and the diagnostic value of each when it occurs in isolation are elaborated. The diagnostic and therapeutic consequences are discussed. PMID- 3024206 TI - Hepatic tumors: US contrast enhancement with CO2 microbubbles. AB - Enhancement of hepatic tumors on sonograms by injection of carbon dioxide microbubbles into the hepatic artery as a contrast material (enhanced ultrasonography) was performed in 43 patients with various histologically confirmed hepatic tumors. Enhanced sonograms were classified into five patterns according to the relative changes of the echo levels between the tumor and the nontumorous parenchyma of the liver as a result of enhancement: hyperechoic change, isoechoic change, hypoechoic change with hyperechoic rim (rim sign), marginal spotty hyperechoic change, and internal spotty hyperechoic change. Eighty-eight percent of hepatocellular carcinomas showed hyperechoic change, 70% of metastatic tumors exhibited hypoechoic change with the rim sign. The marginal spotty hyperechoic change and the internal spotty hyperechoic change were specific for cavernous hemangioma and fibrous granuloma, respectively. This method of enhancement is useful in assessing the nature of liver tumors and in the detection of small nodules in the liver. PMID- 3024207 TI - Portal hemodynamics in patients with hepatocellular carcinoma. AB - The protal blood flow was assessed in 46 patients with hepatocellular carcinoma, 81 with cirrhosis, and 110 control subjects using an ultrasonic B-mode pulsed Doppler duplex system. The cross-sectional area of the portal vein was increased, and the velocity of portal blood flow was decreased in hepatocellular carcinoma and cirrhosis, whereas the blood flow volume was not significantly different. A significant decrease in portal blood flow was found in hepatocellular carcinoma only when at least three of the four major branches of the portal vein were occluded. The change in portal hemodynamics before and after transcatheter arterial embolization (TAE) was investigated. Immediately after TAE, neither portal venous pressure nor portal blood flow showed any constant trend. The portal blood flow reached a peak 1 week after TAE and then returned to its former value after 3-4 weeks, while all cases with poor prognoses showed a drop in portal blood flow after TAE. PMID- 3024209 TI - Clinically occult breast lesions: localization and significance. AB - From early 1974 through September 1985, 927 needle-guided breast biopsies were performed for clinically occult breast lesions. Two hundred and seventy (29%) of these lesions were malignant. This frequency of malignancy was comparable to a 20% frequency in biopsy samples obtained because of clinical (palpable) findings. Of 142 patients with nonpalpable invasive lesions who underwent axillary dissection, 42 (30%) had histologically confirmed axillary metastases. Although these invasive lesions may be clinically occult, they are not inconsequential and demand appropriate treatment. For those patients in this series undergoing mastectomy, the frequency of multicentric cancer in the same breast was 39%. The past 10 years have witnessed a trend in surgical management away from the standard radical or modified radical mastectomy toward limited surgery and radiation therapy for so-called early breast cancer. When planning treatment for these occult lesions, one must consider their biological potential to metastasize as well as their propensity for multicentricity. PMID- 3024208 TI - Pitfall in MR imaging of lymphadenopathy after lymphangiography. AB - In two patients examined with magnetic resonance (MR) imaging after lymphangiography, opacified pelvic lymph nodes could not be distinguished from subcutaneous or retroperitoneal fat because of the short T1 and long T2 relaxation times of lymphangiographic contrast media. Opacified nodes removed from one patient had relaxation times similar to those of fat. Thus, assessment of lymphadenopathy with MR imaging should be performed before lymphangiography to obviate this potential pitfall. PMID- 3024210 TI - Hypervascular hepatic metastases: CT evaluation. AB - Recent reports have indicated that non-contrast material-enhanced scans are of less value than urographic contrast material-enhanced studies in the computed tomographic (CT) evaluation of hepatic metastases. The authors retrospectively reviewed the CT scans of 28 patients with hypervascular liver metastases to determine whether these metastases were more likely to become isodense with the liver after contrast material enhancement, thus necessitating the performance of non-contrast-enhanced scanning. Non-contrast-enhanced and contrast-enhanced incremental dynamic scanning was performed in patients with proved liver metastases from carcinoid tumors (13 patients), islet cell neoplasms (ten patients), pheochromocytomas (four patients), or renal cell carcinoma (one patient). Eleven of the 28 patients (39%) had metastases with non-contrast liver to-lesion attenuation differences greater than 15 HU. These metastases subsequently became isodense or nearly isodense on contrast-enhanced scans. The authors conclude that non-contrast-enhanced CT scanning should be performed in patients with suspected liver metastases from tumors that are usually hypervascular. PMID- 3024211 TI - Tumor thrombus of the inferior vena cava secondary to malignant abdominal neoplasms: US and CT evaluation. AB - Nineteen patients with histologically proved tumor thrombi of the inferior vena cava (IVC) secondary to abdominal neoplasms were studied with the use of ultrasonography (US) and computed tomography (CT). The primary neoplasms were renal cell carcinoma (13 cases), adrenal tumors (two cases), retroperitoneal tumors (two cases), and hepatic tumors (two cases). A positive diagnosis of tumor thrombus was achieved by both methods. The cranial extent of the tumor thrombus was better demonstrated by US studies, which showed echogenic endoluminal material within an enlarged IVC with a bulging anterior wall. On CT scans the tumor thrombus usually appeared as an endoluminal filling defect surrounded by a rim of contrast material. Tumor thrombus was better outlined by CT, particularly when it extended beyond the limits of the IVC wall, as in the retroperitoneal tumors. Neither method could accurately be used to predict IVC wall infiltration when the tumor thrombus remained within the confines of the IVC, nor could either method differentiate tumor from nontumor thrombi. US and CT are complementary in the preoperative assessment of tumor thrombus, and their use obviates the need for venacavography in many cases. PMID- 3024212 TI - [Porcelain veneers--New York University system]. PMID- 3024213 TI - [Advancing periodontitis and root caries]. PMID- 3024214 TI - [Composites--state of the art]. PMID- 3024215 TI - [Macroscopic study of the temporomandibular joint in 5 elderly patients]. PMID- 3024216 TI - [Bacteria causing endodontic complications in restored teeth]. PMID- 3024217 TI - [Non-gamma 2 amalgam in the 1st and 2d generation]. PMID- 3024218 TI - [Kuwata technic. 2]. PMID- 3024219 TI - [Proximal margins of Class II restorations as seen in scanning electron microscopy]. PMID- 3024220 TI - [Occlusal grinding and muscle activity]. PMID- 3024221 TI - [Comparison between palladium alloys and non-precious metal alloys]. PMID- 3024222 TI - Pituitary regulation in endogenous depression. PMID- 3024223 TI - Neuropeptides and cognitive disorders. PMID- 3024224 TI - Pituitary proopiomelanocortin peptides in mental disorders. PMID- 3024225 TI - Anxiety and the benzodiazepine receptor. PMID- 3024226 TI - The design of antagonists of regulatory peptides, and comments on their specificity. PMID- 3024227 TI - Studies on peptide comodulator transmission. New perspective on the treatment of disorders of the central nervous system. PMID- 3024228 TI - Feeding and taste. PMID- 3024229 TI - [The structure of the oncogenic herpesvirus genome and its expression]. PMID- 3024230 TI - [Reconstitution of terminal oxidases of Escherichia coli respiratory chain and its bioenergetics]. PMID- 3024231 TI - [Regulation of cell proliferation by ras gene products through the cAMP cascade system]. PMID- 3024232 TI - [Expression of human c-myc oncogene in prokaryotic and eukaryotic cells]. PMID- 3024233 TI - [The erbB-related growth factor receptors]. PMID- 3024234 TI - [Transforming genes of DNA tumor viruses SV 40, polyoma virus, and adenoviruses]. PMID- 3024235 TI - PAF antagonists: different effects on platelets, neutrophils, guinea pig ileum and PAF-induced vasodilation in isolated rat lung. AB - The effects of structurally different PAF receptor blockers were investigated in platelets, neutrophils, guinea pig ileum, rat isolated lung and rat isolated pulmonary artery. PAF caused serotonin release from platelets and a characteristic shape change and adhesion of neutrophils. The antagonists (CV 3988, alprazolam, 48740 RP and Merck-Sharp and Dohme L-652, 731) inhibited platelet serotonin release but not neutrophil shape change adhesion or lysosomal enzyme release. The antagonists in high concentrations (10(-5)-10(-4)M) inhibited nonspecifically the PAF-induced (10(-8)M) guinea pig ileum contraction, but were ineffective at concentrations which inhibited platelet responses. In the rat lung the compounds, in high concentrations, partially inhibited the low dose PAF induced pulmonary vasodilation and the high dose PAF induced pulmonary vasoconstriction and edema. Our data indicate that some platelet PAF antagonists may be ineffective in blocking the action of PAF on neutrophils and smooth muscle preparations and suggest either PAF-receptor independent actions of PAF or different classes of PAF receptors. PMID- 3024236 TI - Profiles of eicosanoid production by superficial and proliferative colonic epithelial cells and sub-epithelial colonic tissue. AB - The profile of cyclooxygenase and lipoxygenase products in normal rat colonic epithelium and subepithelium was examined. Colons were thoroughly perfused to eliminate contamination with blood. Two preparations of colonic epithelium were employed. The first consisted of intact colonic crypts and epithelial sheets. The second yielded single cell suspensions of superficial versus proliferative epithelial cells. Lipoxygenase product formation by colonic epithelium as measured by hydroxyeicosatetraenoic acid (HETE) and leukotriene B4 (LTB4) production (5-HETE greater than 12-HETE greater than 15-HETE greater than LTB4) accounted for 58% of the total colonic production of these moieties, whereas epithelium accounted for only 20% of total colonic protein. By contrast, prostaglandin (PG) E2 and PGF2 alpha production occurred predominantly (greater than 97%) in the subepithelial layers. The present studies also demonstrate markedly higher levels of accumulation of lipoxygenase products in proliferative versus superficial epithelial cells, whereas prostaglandin accumulation was greater in superficial cells. Previous studies have supported a role for lipoxygenase and cyclooxygenase products in the control of colonic secretion, inflammatory cell infiltration and proliferative activity. The present results raise the possibility that the striking differences in the sites of production of these products within the colon has functional implications. PMID- 3024237 TI - [A case of basal cell carcinoma of the tip of the nose in the form of a pedunculated tumor]. PMID- 3024238 TI - Atrial natriuretic factor-induced cGMP accumulation in rat anterior pituitary cells in culture is not coupled to hormonal secretion. AB - While atrial natriuretic factor (ANF) does not influence ACTH secretion, it was reported to have a marked stimulatory effect on the intracellular accumulation of cGMP in rat anterior pituitary cells in culture. Since many biological actions of ANF appear coupled to its excitatory action on target cell guanylate cyclase, the current study was designed to characterize the ANF-induced cGMP response in anterior pituitary with a view to determining whether the nucleotide plays a regulatory role in the secretory function of this gland. A 3 min exposure of cells in primary culture to 300 nM ANF (99-126) or 100 microM sodium nitroprusside (SNP), a stimulator of guanylate cyclase, causes maximal 10- and 3 fold elevations of cGMP levels, respectively. Following a progressive decrease, 6 and 2-fold increases over basal cGMP levels are still observed after 180 min of incubation with ANF (99-126) and SNP, respectively. The half-maximal stimulation of cGMP accumulation induced by a 10 min exposure to ANF (99-126), or rat atriopeptin II (ANF 103-125) is observed at 9 +/- 2 and 125 +/- 22 nM, respectively. ANF fragments (99-109) and (111-126), as well as human cardiodilatin (hANF 1-16), do not alter cGMP levels. Basal and ANF-induced cGMP levels are at least 10-fold higher in cell populations enriched in gonadotrophs compared to gonadotroph-impoverished preparations. A 3 h incubation of cells with ANF (0.1-1000 nM), however, fails to modify spontaneous or LHRH-induced LH secretion. Similarly, ANF does not alter spontaneous release of GH, TSH or PRL. The data suggest indirectly that gonadotrophs represent a principal site at which ANF acts to stimulate cGMP synthesis, but that the nucleotide is not a specific regulator of the LH secretory process; nor is it generally involved as a second messenger in the secretory function of any cell type of the anterior pituitary gland. PMID- 3024239 TI - [Fundamental and clinical studies on the measurement of beta 2-microglobulin as a tumor marker using the Dainabot beta 2-micro-radioimmunoassay kit]. PMID- 3024240 TI - [Determinative technics of plasma and salivary cortisol in patients with chronic nephropathies in hemodialysis programs]. PMID- 3024241 TI - [Effect of the combination of cyproterone acetate and conjugated estrogens on hirsutism, alopecia and acne]. PMID- 3024242 TI - [Fibrolamellar hepatocarcinoma with a long clinical course and possible production of fibrinogen]. PMID- 3024243 TI - [Double-contrast pharyngography in surgically treated and/or irradiated oro- and hypopharyngeal malignant tumors]. AB - The use of a double contrast technique in the oropharynx and hypopharynx is described and its diagnostic value following treatment is defined. By means of the double contrast pharyngogram, it is possible to demonstrate the pharyngeal mucosa and, post-operatively, the anastomosis. It is the only imaging method which demonstrates the course of swallowing and passage through the pharynx. By paying attention to physiological changes following treatment and taking account of early post-therapeutic appearances, it is possible to form a judgement concerning the peripharyngeal tissue. CT, however, is the method of choice for demonstrating an extraluminal tumour. In view of the ease of the method, double contrast pharyngography is the ideal radiological procedure for close follow-up of patients with malignant tumours of the pharynx. PMID- 3024244 TI - [Nuclear magnetic resonance tomographic staging of tumors of the oral cavity, oro and hypopharynx and the larynx. A comparison with computed tomography and sonography]. AB - Thirty-six patients with tumours of the mouth, the oropharynx and hypopharynx and the larynx were examined by nuclear resonance tomography. The results were compared with the clinical findings of inspection and palpation and with CT and sonography, with respect to T and N classification. In seven patients the classification could be confirmed at operation. NMR provides very good anatomical detail and marked contrast between tumour and the surrounding tissues, particularly on T2 weighted images. NMR showed the best correlation with the clinical findings as regards the T classification and was the most accurate method, as confirmed by surgery. It is superior to CT and sonography for diseases in the oropharynx and hypopharynx. For the examination of the cervical lymphatics, sonography remains the recommended method. PMID- 3024245 TI - [Bronchioalveolar carcinoma]. AB - A study of 21 patients suffering from bronchioalveolar carcinoma and 1028 comparable cases of the literature is presented. By this we gained statistical data concerning radiological and clinical manifestations of this rare tumour. However, radiological signs estimated to be typical of this carcinoma, like "rabbit ear sign" and positive "air bronchogram" are too rare to be really helpful for putting up the diagnosis. At time of diagnosis the mean age was 58 years. Men are hardly more frequently concerned than women. Smoking seems to have no aetiological significance. The most common clinical symptom was chronic cough followed by chest pain. A large amount of sputum--a symptom which sometimes has been estimated to be typical of this tumour--occurred only very rarely. PMID- 3024246 TI - [Causes and x-ray symptomatic types of isolated bronchoceles]. AB - A survey is given of the roentgenographic features of isolated bronchoceles (25 cases: 9 men; 16 women). If some roentgenomorphological peculiarities are taken into consideration, identification is possible via plain radiography and topic related two-dimensional tomography. With regard to preoperative aspects the perception of this disease is very important because the extent of a surgical procedure may not infrequently be limited to a segmental resection. PMID- 3024247 TI - [Computed tomography of lung contusion. An experimental study]. AB - A new experimental model has been developed to determine whether computed tomography has any advantages for the investigation of lung contusions. In 27 dogs, chest wall deformation was produced at speeds varying from 17.3 m/sec to 45.7 m/sec, leading to lung contusion. After the injury, 27 out of 27 (100%) CT examinations demonstrated the contusions, but only nine out of 24 (37.5%) were seen on conventional radiographs. In contrast to conventional radiography, which either failed to show the presence of a contusion, or which under-estimated the extent by more than one cm, in more than half the cases CT agreed with the pathological findings in more than 90%. No lung contusions were found at autopsy which had not been demonstrated by CT. PMID- 3024248 TI - [Accuracy of cardiac CT and echocardiography in the diagnosis of space-occupying processes in the heart]. AB - CT of the heart and echocardiography was performed on 107 patients with various space-occupying lesions of the heart. In addition, 27 patients were subjected to angiocardiography. A comparison of the findings has shown that cardio-CT is superior to two-dimensional echocardiography for demonstrating pericardial masses and intracavity thrombi. There was no significant difference as far as intracavity or intramural tumours were concerned. CT and sonography are superior to angiography in the diagnosis of cardiac space-occupying lesions. PMID- 3024249 TI - [Single-catheter technic for intra-arterial digital subtraction angiography of the aortic arch and selective imaging of the supra-aortic arteries]. AB - A double-curve catheter with side-holes was used for intraarterial digital subtraction angiography of the aortic arch and selective angiography of the supraaortic arteries. Applying this technique the same catheter can be used in the majority of cases (approximately 75%) if aortic arch angiography is followed by selective catheterisation of the supraaortic arteries. PMID- 3024250 TI - Non-invasive evaluation of prosthetic dialysis shunts in asymptomatic patients. AB - A prospective study was done in 16 asymptomatic patients on chronic haemodialysis after implantation of a PTFE vascular access. A high percentage (50%) of these grafts turned out to be stenotic somewhere along their course. All these stenoses were detected by real-time ultrasound (accuracy 100%). Accuracy for Doppler examination was less, being 95% for spectral broadening and 98% for increased systolic peak flow frequency. So real-time ultrasound with or without Doppler examination is the preferable method for early detection of stenosis in PTFE dialysis shunts in asymptomatic patients. PMID- 3024251 TI - [Pulsed duplex sonographic follow-up of varicoceles following sclerotherapy]. AB - Forty-one patients were examined by pulsed duplex sonography immediately before and then one, four and twelve weeks following sclerotherapy of a varicocele. The severity of the varicocele was determined by sonography before treatment and confirmed by phlebography. Sonographic follow-up showed reduction, or disappearance, of the varicocele in 78% of cases. Doppler examination confirmed successful treatment in 85% of patients. Pulsed duplex sonography, combining a non-invasive imaging method and a functional technique, is well suited for the follow-up of varicoceles after treatment. PMID- 3024252 TI - [Gastrointestinal amyloidosis as a differential diagnostic problem]. AB - The diagnostic radiologist may have problems with the differential diagnosis of gastrointestinal amyloidosis combined with only uncharacteristic clinical symptoms. In the stomach upper gastrointestinal series show in most cases stenosing submucosal masses in the gastric antrum with diminished peristalsis and pliability. Sonography reveals a circular thickening of the gastric antrum wall. Only a synopsis of radiologic changes, the patients's history, laboratory tests and biopsies render a clue to the correct diagnosis. In the small bowel segmental or total intestinal dilatation with sonographically demonstrable thickening of the bowel wall and diminished motility, prolonged transit and eventually obstruction or paralytic ileus can be demonstrated. In patients with simultaneous plasmocytoma the radiologist has to take a gastrointestinal involvement by concurrent amyloidosis into account. PMID- 3024253 TI - [Radiological diagnosis of aggressive fibromatosis]. AB - Aggressive fibromatosis (desmoid, desmoid tumour) resembles, in its infiltrating and destructive growth, a fibrosarcoma, but does not metastasize. Because of its high recurrence rate, the tumour remains a surgical problem. Various imaging methods were evaluated retrospectively in 23 patients with histologically confirmed aggressive fibromatosis. Conventional radiological procedures are poor at demonstrating the extent and type of tumour. Modern tomographic methods are more able to determine the size of the lesion and a combination of angiography and CT can frequently provide a definite diagnosis. PMID- 3024254 TI - [Diagnosis of lymphomas on the MR tomogram]. AB - MR tomography provides images equal to, or superior to, CT of lymph node involvement in the thorax or abdomen. It may be possible to achieve tissue characterisation by means of T1 and T2 images. Chronic inflammatory diseases causing lymph node enlargement, such as sarcoidosis, show characteristic T1 intervals (about 650 msecs), whereas lymphomas show intervals of 900 msecs. Enlarged lymph nodes of low malignancy show lower T2 values (78 to 94 msecs), highly malignant show higher T2 values (83 to 145 msecs). Already we believe MR to be the method of choice in evaluating lymphoma and sarcoidosis. PMID- 3024255 TI - [Use of Gd-DTPA in the nuclear magnetic resonance study of the breast]. AB - 55 patients with altogether 60 breast masses have been examined postoperatively by plain MR, MR with Gd-DTPA and mammography at our institution. All breast carcinomas and fibroadenomas did enhance significantly. Non-proliferative dysplasia, normal breast tissue and scar tissue did not enhance significantly. Borderline enhancement has been found in cases of proliferative dysplasia. The diagnostic accuracy has been improved by MR with Gd-DTPA. Advantages have been an improved visualization of tumors in dense breasts, an improved differentiation between irregular dysplasia and carcinoma and an improved differentiation between post-therapeutic changes (after radiation, surgery or silicon implants) and carcinoma. Nevertheless both technical improvements and further clinical experience is necessary. PMID- 3024256 TI - [MR tomography of hemophilic arthropathy of the knee joint]. AB - The value of MR tomography in the diagnosis and follow-up of haemophilic arthropathy of the knee was studied in 40 examinations, using special surface coils. Good spatial resolution and reconstruction in various planes provide an accurate demonstration of the lesion, particularly of the intra-articular structures. Abnormalities of the synovia and of hyaline cartilage can be demonstrated. MR tomography is an important imaging method which is superior to sonography or CT in demonstrating complications resulting from the bleeding, such as effusions and haemarthroses. PMID- 3024257 TI - Diagnosis of osteoid osteoma by digital subtraction angiography. AB - The purpose of this paper is to stress the importance of DSA in the diagnosis of osteoid osteoma. Three cases are reported in which DSA with arterial injection of contrast material demonstrated a typical nidus enhancement. The major role of DSA in establishing the exact nature and localisation of the disease is discussed. PMID- 3024258 TI - [Diagnosis of instability of the upper cervical spine by functional computed tomography]. AB - The evaluation by means of functional x-rays, of rotatory instability of the upper cervical spine as a result traumatic or inflammatory destruction of the ligamentous apparatus, is unsatisfactory. Functional CT of the upper cervical spine allows measurement of the segmental rotatory movements. 9 healthy adults and 30 patients were examined after neck injury via functional CT's. A rotation between occiput and atlas greater than 9 degrees, between atlas and axis over 50 degrees, the left-right difference at the level C0/C1 greater than 6 degrees and at the level C1/C2 over 10.5 degrees point to a suspicion of hypermobility or instability. PMID- 3024259 TI - [The value of early postoperative CT study after lumbar intervertebral disk surgery]. AB - The changes of the early intraspinal conditions few days after an operation of a lumbar disc herniation are little known. We, therefore, examined these operation areas by CT on a representative group of 256 patients. Besides the "common" findings, such as seroma, one must classify conditions such as haematomas, isolated bone fragments and/or residual disc tissue as "unusual" changes. The CT findings are correlated with clinical symptoms. We conducted CT and clinical follow-up studies in 91 patients several weeks later. All patients showed epidural scars of very different intensity. The comparison between early postoperative findings and the controls documents the high accuracy of the interpretation of the early postoperative states seen by CT. Therefore, it is possible to give hints for the indication for early postoperative CT examinations, for the prognosis as well as for the technique for a safe operative procedure. PMID- 3024260 TI - [Cystic changes of the posterior cranial fossa on the nuclear magnetic resonance tomogram]. AB - MR images were obtained in 15 patients with cystic lesions in the posterior fossa and the results compared with computed tomography (CT). The abnormalities imaged included 5 epidermoids, 3 haemangioblastomas, 4 arachnoid cysts, 1 postoperative cyst, and 4 large cisterns. One advantage of MR is the ability to demonstrate lesions in three planes and without artifacts. Liquor-related intensity measurements of cyst contents provided important information, thereby improving diagnostic specificity and patient management. T2-weighted images were found to be more sensitive in characterising cystic lesions of the posterior fossa than were T1-weighted images. Large cisterns, arachnoid and postoperative cysts had an intensity pattern identical or almost identical to cerebral spinal fluid. More proteinaceous cysts, including epidermoids and to a higher degree haemangioblastomas, had clearly higher intensity than cerebral spinal fluid on the T2-weighted images. PMID- 3024261 TI - [Absolute measurement of laminar flow with the aid of an orthogonal excitation technic in NMR tomography]. AB - A method for absolute measurement of flow quantities by excitation of a slice orthogonal to the measuring plane is presented. The developing flow profile can be imaged directly and its dynamic behaviour can be sampled and measured using the multiecho technique. Simple formulas can be derived by means of Hagen Poiseuille's law for quantification. PMID- 3024262 TI - [Biloma in the subhepatic space and omental bursa after revision of the choledochus]. PMID- 3024263 TI - [Asymptomatic duplication of the small intestine--a rare cause of Chilaiditi's syndrome]. PMID- 3024264 TI - [Asymptomatic giant emphysematous bulla of the lung]. PMID- 3024265 TI - [Histological confirmation of monostotic Paget's disease in the sacrum by CT guided bone puncture biopsy]. PMID- 3024266 TI - Traumatic lumbosacral nerve root avulsion demonstrated via computerised tomography. PMID- 3024267 TI - [Cerebellar hemangioblastoma (Lindau tumor) in MR tomography]. PMID- 3024268 TI - [Etomidate: a questionable drug]. PMID- 3024269 TI - [Cytological and ultrastructural diagnosis of human polyoma virus infection (BK) in urinary sediment]. PMID- 3024270 TI - [Endogenous opiates and essential obesity]. PMID- 3024271 TI - [The problem of the addictive capacity of zipeprol. Results of an experimental and clinical investigation]. PMID- 3024272 TI - Papaverine inhibits binding of [3H]-prazosin and [3H]-yohimbine to rat renal cortical membranes. AB - We examined the effects of papaverine on specific [3H]-prazosin and [3H] yohimbine bindings to rat renal cortical membranes and compared these effects with those of verapamil. Papaverine inhibited both ligand bindings with KI values of 4.2 +/- 1.7 microM and 48.4 +/- 8.7 microM (N = 5) in the inhibition of [3H] prazosin and [3H]-yohimbine bindings, respectively. In contrast with verapamil, which competitively inhibited both ligand bindings, papaverine inhibited them in a different manner. Although the affinities of these ligand binding sites for papaverine are low, alpha-receptor blockade is a possible mechanism of action of this compound. PMID- 3024273 TI - Reversible down-regulation of liver cyclo(His-Pro) binding sites by cyclo(His Pro) administration to mice. AB - Intraperitoneal injections of histidyl-proline diketopiperazine [cyclo(His-Pro), 15 mg/kg/day] to mice for 3 days caused a significant loss of cyclo(His-Pro) binding sites in the liver. This loss depended on the decrease in the number of binding sites and not on the change in affinity. The cyclo(His-Pro)-mediated loss was reversible, and the specific binding returned to the original level within 3 days of the cessation of cyclo(His-Pro). These results suggest that cyclo(His Pro) participates in regulating its own binding sites in the liver. PMID- 3024275 TI - Interactions between local and generalised burn edema. AB - In judging the infusion therapy of burn injuries, not only the circulatory and pulmonary effects are considered but also the local effect on burned tissue. Local edema leads to hypoxia as well as to the burn-specific sludge phenomenon. Microcirculation and edema are often adversely influenced by our therapeutic efforts. Therapeutic proteinase inhibition, for example, has a beneficial effect on edema formation but reduces spontaneous fibrinolytic activity and thus increases the sludge phenomenon. Standardized experimental scalds of 30% of the body surface in rabbits increased density of the lungs, as determined by computed tomography. Colloidal infusion therapy was found to diminish fluid shift into the lung more than crystalloid infusion therapy. However, all tested infusion regimens, using identical quantity, increased edema formation in the burned skin. Biseco (natural serum proteins containing igG, igA and igM; Fa. Biotest, Frankfurt, F.R.G.) and Solcoseryl (a protein-free extract of calf blood stimulating the oxygen efficiency; Fa. Solco Basel) were demonstrated to diminish local edema formation as well as lung edema secondary to burn injuries. PMID- 3024274 TI - Comparison of the inhibitory effects of aspirin and ticlopidine on platelet aggregation and Ca2+ mobilization in rat platelets. AB - To study the cause of the sex difference in the inhibitory effects of aspirin on platelet functions, we compared the effects of aspirin and ticlopidine in rat platelets. Aspirin showed the sex difference in the effects on the collagen induced platelet aggregation and the malondialdehyde (MDA) production, but ticlopidine showed no sex difference in the effects on those. The C-AMP level in male rat platelets was the same as that in female rat platelets. Ticlopidine increased the C-AMP level in rat platelets, but aspirin did not. Aspirin suppressed the intracellular Ca2+ mobilization in male rat platelets more strongly than in female rat platelets. Ticlopidine suppressed the Ca2+ mobilization in both sexes equivalently. The above results suggest that the sex difference in the inhibitory effects of aspirin on rat platelet functions comes from the sex difference in the thromboxane (TX) production in rat platelets. PMID- 3024276 TI - Generation of mediators by limited proteolysis during blood coagulation and fibrinolysis--its pathogenetic role in the adult respiratory distress syndrome (ARDS). AB - Products of blood coagulation and fibrinolysis, which are generated by limited proteolysis are able to effect the pulmonary circulation and gas exchange by biochemically mediated actions. Thrombin, fibrin and its degradation products provoke functional and morphological changes in the vessel wall. Fibrinopeptides, fibrin monomers and fibrin (ogen) degradation products induce vasoconstriction and vascular leakage. The vasoconstricting action of fibrin monomers is mediated by thromboxane A2 (TXA2) which is synthetized in the lung tissue itself. Thromboxane synthesis is stimulated by fibrin monomers only in the pulmonary circulation and not in the systemic circulation. Besides their vascular effects, fibrin monomers disturb the function of the surfactant components and increase the surface tension in surfactant monolayers. Proteinase inhibition as a general prophylactic and therapeutic concept with adult respiratory distress syndrome (ARDS) has to include the system of blood coagulation and fibrinolysis predominantly to prevent or stop the generation of products which are able to induce pulmonary vasoconstriction, vascular leakage and impairment of pulmonary gas exchange. PMID- 3024277 TI - The proteinases in shock. AB - The nomenclature of proteinases and their role in shock are reviewed. The regulation of the kallikrein-kinin system, coagulation and fibrinolysis is described with special emphasis on the role of granulocytes for the proteolysis in shock particularly in the lung. The effect of proteinase inhibitors on such systems is described, together with proteinase inhibitors effects on granulocytes and their possible role in the prevention especially of lung damage after shock. PMID- 3024278 TI - Uncontrolled plasma proteolysis: a major threat to the septicemic patient. AB - In order to further elucidate the pathophysiological significance of plasma proteolysis during septicemia, surgical patients with septicemia were studied by means of chromogenic peptide substrate assays. In fatal cases continuous low values for prekallikrein, plasminogen and antithrombin III were found until death. At autopsy a persistent septic focus was found in all but one of the fatal cases. Very low levels of prekallikrein during sepsis and reduced functional inhibition of plasma kallikrein in septic shock indicated a poor prognosis. In the survivors the parameters returned towards the normal range upon successful therapy. Furthermore the paper demonstrates the application of a new parameter, the proenzyme functional inhibition index (PFI-index) in patients with septicemia. The data reveal that by means of this parameter patients at high risk can be identified at an early stage of the disease. PMID- 3024279 TI - Etiology of the pulmonary pathophysiology associated with inhalation injury. AB - This study describes an experimental model of smoke inhalation injury in sheep in which the same pathophysiologic alterations occur as with clinical inhalation injury in man. Diffuse pulmonary mucosal sloughing with atelectasis and emphysema with concomitant development of pulmonary edema results in a decrease in arterial oxygen and progressive pulmonary deterioration which results in a substantial mortality. Increased pulmonary edema fluid is shown to be caused by an increased microvascular permeability to protein with pulmonary lymph and tracheobronchial fluid, a filtrate of plasma. Concomitant with this increase in microvascular permeability is an influx of neutrophils, release of proteolytic enzymes and an identified presence of the metabolite of the prostanoid thromboxane A2 which are postulated as contributors to the progressive pulmonary dysfunction post inhalation injury. PMID- 3024280 TI - Continuous intravenous infusion of elastase in normal and agranulocytic minipigs- effects on the lungs and the blood coagulation system. AB - In order to investigate the acute effects on lung morphology, lung function, hemodynamic and blood coagulation system, elastase (330 units (U) kg-1 h-1) was continuously infused into 16 anesthetized and mechanically ventilated minipigs. Elastase infusion induced a disturbance of blood coagulation leading to hypocoagulability, a pulmonary leukostasis, interstitial edema, a progressive respiratory failure with prompt increase in pulmonary vascular resistance, decrease in systemic vascular resistance, increased venous admixture, and increased dead space ventilation. Agranulocytosis prevented interstitial edema but not disturbances in pulmonary or hemodynamic function or hypocoagulability. The results clearly indicate that elastase may be involved in the pathophysiology of acute lung failure and defects in the blood coagulation system. PMID- 3024281 TI - Effect of a proteolytic enzyme inhibitor on the cardiopulmonary response to endotoxin. AB - Gabexate mesilate (GM), a proteolytic enzyme inhibitor, was given to 31 chronically instrumented sheep either as a pretreatment or treatment to determine its effects on the cardiopulmonary response to endotoxin (ET). Twelve sheep received GM prior to endotoxin (0.75 microgram/kg), 10 after ET and 9 received GM without ET. A typical biphasic response was noted; phase I showed increased lymph flow (QL), reduced lymph to plasma protein ratio (L/P) and increased pulmonary artery pressure (PAP). Phase II occurred two hours after ET; the lymph flow remained elevated and the L/P ratio rose slightly above control. Both phases demonstrated an elevation in total peripheral vascular resistance (TPR) and hematocrit but a reduction in cardiac output (CO). In phase II, QL was reduced 35% with pretreatment; the L/P ratio did not rise as much and the PAP was unchanged. There was also a 35% reduction in lymph flow; no change in the lymph to plasma protein ratio but there was an elevation in pulmonary microvascular pressure. Pretreatment with GM diminished the fall in CO and the rise in TPR seen with endotoxin. The values were unaffected by treatment alone. PMID- 3024282 TI - [Percutaneous transluminal angioplasty in venous systems]. PMID- 3024283 TI - Sodium bicarbonate infusion increases discharge frequency of intrapulmonary chemoreceptors only at high CO2. AB - We felt that earlier determinations of independent effects of extracellular pH and PCO2 on intrapulmonary chemoreceptors (IPC) discharge frequency were difficult to analyze because they used perfused lungs, and ventilation-perfusion changes among parabronchi could not be controlled. We decided to repeat these studies in non-perfused lungs. We cannulated both extrapulmonary bronchi of 10 thoracotomized Pekin ducks anesthetized with sodium pentobarbital (25-35 mg/kg) and unidirectionally ventilated each lung. The perfused right lung maintained gas exchange while the non-perfused left lung received 0.6 L/min of CO2 mixed in air. We recorded the discharge frequency of one IPC per duck at various PCO2, re established circulation, and infused 3.0 mmol/kg of sodium bicarbonate intravenously. After 15 min, discharge frequencies were again measured from the same IPC in the nonperfused lung. The slopes and intercepts of discharge frequencies vs ln PCO2 relationship were depressed in six IPC, increased in two IPC and not significantly affected in two IPC. Arterial pH was increased significantly (0.11 unit) at 38 Torr arterial PCO2. We conclude that acutely increased extracellular sodium bicarbonate affects IPC discharge only by depressing sensitivity of most IPC to PCO2 and does not have an independent effect through pH. PMID- 3024284 TI - [Osteomalacia following gastrectomy. A case report]. PMID- 3024285 TI - [Clinico-pathologic conference: hemiplegia and coma in acute lymphatic leukemia]. PMID- 3024286 TI - [Topical treatment of allergic rhinitis]. PMID- 3024287 TI - [Anatomo-clinical conference at the Salpetriere. October 1985. Sensory neuropathy and memory disorders developing slowly in a 65-year-old woman]. PMID- 3024288 TI - Molecular epidemiology of bacterial infections: examples of methodology and of investigations of outbreaks. AB - Plasmid profile analysis, plasmid and chromosomal bacterial restriction endonuclease DNA analysis, and DNA hybridization are increasingly used in clinical microbiology and epidemiology. These techniques have been applied, singly and in combination, to investigations of outbreaks, including those of diarrheal diseases caused by pandemic Vibrio cholerae, enterotoxigenic and enterohemorrhagic Escherichia coli, Salmonella muenchen, and Salmonella typhimurium. The techniques have been critical in the unraveling of outbreaks of disease caused by nosocomial and community-acquired pathogens. Although there are limitations to these methods, their applications have important ramifications for basic science and public health. PMID- 3024289 TI - Plasmid analysis in the study of the epidemiology of nosocomial gram-positive cocci. AB - Gram-positive pathogens remain important causes of nosocomial infection. Plasmid analysis has provided useful information in the study of the epidemiology of these organisms, particularly Staphylococcus epidermidis. The antimicrobial resistance in these organisms seems to parallel that of gram-negative rods, with transposition, plasmid exchange, and strain dissemination interacting in the development of resistance. PMID- 3024290 TI - Molecular epidemiology of antibiotic resistance plasmids of Haemophilus species and Neisseria gonorrhoeae. AB - Ampicillin resistance in Haemophilus influenzae and Neisseria gonorrhoeae is most commonly due to plasmid-mediated production of the TEM beta-lactamase. The H. influenzae plasmids may have evolved by insertion of various antibiotic resistance transposons into a phenotypically cryptic plasmid found in one of 699 isolates of H. influenzae examined. The small, nonconjugative, beta-lactamase specifying plasmids of N. gonorrhoeae and Haemophilus species are highly related. Phenotypically cryptic plasmids found in several epidemiologically distinct isolates of Haemophilus parainfluenzae are highly related to the beta-lactamase plasmids but carry no transposon A (TnA) sequences. This evidence strongly favors the hypothesis that the beta-lactamase plasmids evolved by the insertion of TnA (possibly introduced from enteric bacteria) into cryptic plasmids resident in H. parainfluenzae. PMID- 3024291 TI - The changing pattern of trimethoprim resistance in Paris, with a review of worldwide experience. AB - From 1972 to 1984, all Enterobacteriaceae isolated from clinical specimens at St. Joseph Hospital in Paris were tested for susceptibility to trimethoprim. During this period, resistance to trimethoprim increased from 17.9% to 25.5%; the increase was due mainly to strains with high-level resistance. Genetic studies, including transferability, incompatibility grouping, determination of the molecular mass of plasmids, and hybridization with dihydrofolate reductase I and II genes, were performed with randomly selected strains, and the results were compared with those of similar studies in other countries. The most striking phenomenon in trimethoprim-resistant strains was the presence of various resistance mechanisms and of different plasmids, transposons, and genetic determinants. PMID- 3024292 TI - Controversies in viral diagnosis. AB - A number of fallacies concerning viral diagnosis persist. It is said that viral studies are useless since there is no treatment for viral diseases, but suitable therapies are available for a number of severe viral infections. Viral culture results are thought to be too slow to affect patient management, but the average time for detection of a virus is three to four days, and greater than 70% can be reported within five days; detection of viral antigen can be provided in hours. It is thought that viral infections are best diagnosed by serologic tests. However, serologic tests requiring demonstration of a rise in titer or seroconversion are inherently slower than cultures and therefore less useful. Genetic probes are insensitive compared with cultures, and enzyme-linked immunosorbent assay methods for detecting viral antigen are no more sensitive than fluorescent antibody techniques and do not provide a means of assaying the quality of the sample. Finally, it is often stated that viral studies should be performed only in reference or regional laboratories. Regional virus laboratories should be located in teaching hospitals, and networks should be established to permit hospital laboratories to be involved at different levels. PMID- 3024293 TI - [Inoperable non-small cell bronchopulmonary cancers. Results of ambulatory polychemotherapy including cisplatin]. AB - Between October, 1981 and October, 1983, 57 subjects with inoperable non small cell carcinoma of the lung (localized in 37, metastatic in 20) were treated as out-patients, after oral rehydration, with a multiple chemotherapy regimen which included Cisplatinum, Doxorubicin, Vincristine and Lomustine. No severe haematological or renal reaction was observed. After 2 courses 23 patients were responders (complete, or partial, or minor response), 2 of them with complete response. Patients with localized tumour had complementary radiotherapy, while chemotherapy was pursued in responsive patients with metastasis. Median survival was 9 months in responders and 5 months in non-responders. Thus, chemotherapy including Cisplatinum in medium doses (80 ms/sq.m) can be given to out-patients without risk of renal damage. The results obtained in this study are comparable to those of studies performed with other drug combinations including Cisplatinum. PMID- 3024294 TI - [Mode of action of corticoids in asthma]. AB - Generally speaking, corticosteroids exert their effects on cells through genomic and non genomic mechanisms. They both suppress and enhance numerous biological phenomena. In asthma, corticosteroids act at different levels. They: inhibit chemical mediators, whether these are performed, like histamine, or newly formed, like arachidonic acid metabolites (prostaglandins and leukotrienes) or the platelet-activating factor (PAF); restore the sensitivity of beta-adrenergic receptors to sympathomimetic drugs; exert a powerful anti-inflammatory effect, notably reducing bronchial mucus secretion; reduce bronchial hyperreactivity and modify the bronchial response to bronchoconstrictors; act on respiratory function and gas exchanges. Many effects of corticosteroids in asthma involve the synthesis of proteins, such as lipomodulin (or macrocortin) which inhibits phospholipase A2, a key-enzyme in the synthesis of numerous chemical mediators derived from membrane phospholipids. The multiple effects of corticosteroids account for their broad spectrum of activity and their effectiveness against both acute and chronic manifestations of asthma. PMID- 3024295 TI - [Association of a granular cell tumor with a hamartochondroma]. PMID- 3024296 TI - [Parvovirus B 19 infections]. AB - We report the human parvovirus (HPV) infection cases diagnosed in 1984 and 1985 in the virological laboratory of C.N.T.S. Detection of viral antigen and total anti-HPV antibodies was performed by electroimmunodiffusion, and specific IgM by antibody-capture-radioimmunoassay. Seven viraemias were found in 38,730 sera sent for detection of hepatitis A and B markers. Twenty-two observations of aplastic crisis are described, with underlying haemolytic anaemias in 21 cases. HPV infection was serologically proven in 4 cases out of 22 rubeolelike illness, and, in 17 cases out of 17 erythema infectiosum (fifth disease). Four cases of vascular purpura (one of which was Schonlein-Henoch purpura) were studied, and HPV antigen was isolated in 2 cases. An observation of arthropathies in a young adult is described. Finally, 2 spontaneous abortions were simultaneous to an HPV infection. Our study allows us to underline the following points: HPV is the principal (but not exclusive) agent of the aplastic crisis in chronic haemolytic anaemias; these acute erythroblastopenias can reveal an unknown haemolytic anaemia, in particular in hereditary spherocytosis; the polymorphism of the clinical expression of HPV infection is important. Some viraemias are asymptomatic, while others are accompanied by vascular purpura, or atypic erythema; HPV has never been isolated in the fifth disease. Nonetheless bringing out of specific IgM and association of HPV infection linked manifestations in a same patient or in a same family shows its responsibility in the fifth disease; HPV responsibility in spontaneous abortions and fetal malformations remains to be demonstrated. PMID- 3024297 TI - Viral antibodies in serum and cryoprecipitate of patients with essential mixed and secondary cryoglobulinemia. Preliminary results. AB - Antibodies to measles virus, CMV, HSV1, HSV2 and EBV have been tested in 9 patients with essential mixed cryoglobulinemia (EMC) and 24 patients with cryoglobulinemia associated with lymphoproliferative, autoimmune or hepatic diseases (secondary cryoglobulinemia, SC). The assays were performed in serum, cryoprecipitate and supernatant by using immunoenzymatic methods. The highest prevalence was observed in the supernatant of both EMC and SC patients. Antibodies to measles virus show a serum/cryoprecipitate ratio higher than that of the other viral agents. Statistically significant differences between EMC and SC patients in the positivity of the various viral antibodies were not shown. PMID- 3024298 TI - Chronic-relapsing polyneuropathy in the course of cryoglobulinemia. Clinical aspects and plasmapheretic treatment. AB - During the last 3 years we have observed 7 cases (4.1%) of cryoglobulinemia associated polyneuropathy out of 167 patients with demonstrated cryoglobulinemia. Nerve involvement in the course of cryoglobulinemia is usually symmetrical, affecting the distal portions of the limbs and has a chronic or chronic-relapsing evolution. In our experience, the combined treatment by plasmapheresis and glucocorticoids and/or cytostatic drugs has proved to be quite effective, particularly when instituted during the symptomatic exacerbations. Therefore, we suggest to perform a systematic investigation of the cryoglobulinemic phenomenon in every patient presenting with symptoms and/or signs of an apparently idiopathic polyneuropathy. PMID- 3024299 TI - Plasma-exchange in mixed cryoglobulinemia. Effects on renal, liver and neurologic involvement. AB - Prolonged plasma-exchange without addition of cytotoxic agents was employed in 16 patients with mixed cryoglobulinemia and kidney, liver or neurologic involvement. Patients with rapidly progressive renal failure or active and reversible lesions generally improved after plasma-exchange, as well as those with a recently occurring sensory-motor peripheral neuropathy. In 4 out of 6 patients with mixed cryoglobulinemia and chronic active hepatitis, plasma-exchange was followed by either normalization or significant reduction of liver enzymes and bromosulfophthalein retention. In all cases responding to plasma-exchange the beneficial effects were evident after the first 2-3 weeks of treatment, while symptoms did not generally recur when the procedures were either slowly tapered or discontinued. Although the pathogenetic mechanism(s) of action of plasma exchange remains largely unknown, preliminary data indicate that these procedures induce quantitative as well as qualitative changes in the immune system. PMID- 3024300 TI - [Dietetic fiber and constipation]. PMID- 3024301 TI - Mammary gland tumors in irradiated and untreated guinea pigs. AB - This is a report of mammary gland tumors from 62 guinea pigs. The tumors arose in the terminal ductal-lobular units as either lobular acinar carcinoma or cystadenocarcinoma or as papillary carcinomas within large ducts near the mammilla. About half the number of the males had terminal ductal-lobular carcinomas and all but 2 of the papillary duct carcinomas also arose in males. Large tumors frequently exhibited squamous, chondromatous, osseous, fatty and myoepitheliomatous types of tissues. In 2 irradiated males and 1 female the tumors metastasized. Whole-body irradiation did not produce significant changes in the number or sex distribution or in the morphology of mammary gland tumors in inbred or outbred guinea pigs. All females had cystic ovaries without increase in granulosa cells, 24 (66.6%) had uterine tumors and 13 (34.2%) had adrenal gland tumors; all males had atrophic testes, 5 (16.5%) had testicular and 6 (22.2%) had adrenal gland tumors. PMID- 3024302 TI - Inhibition of tumor development by dehydroepiandrosterone and related steroids. AB - The naturally occurring adrenal steroid, dehydroepiandrosterone (DHEA), is a potent non-competitive inhibitor of mammalian glucose-6-phosphate dehydrogenase (G6PDH). Oral administration of DHEA to mice inhibits spontaneous breast cancer and chemically induced tumors of the lung and colon. Topical application of DHEA to mouse skin inhibits 7,12-dimethylbenz(a)anthracene (DMBA)-initiated and tetradecanoylphorbol-13-acetate (TPA)-promoted papillomas and DMBA-induced carcinomas at both the initiation and promotion phase. Evidence is presented that critical steps in the initiation process (mixed-function oxidase activation of a carcinogen) and promotion process (enhanced rates of cell proliferation and superoxide formation) all require NADPH and may be inhibited by DHEA and structural analogs as a result of a lowering of the NADPH cellular pool. Results obtained by others with fibroblasts and lymphocytes from individuals with the Mediterranean variant of G6PDH deficiency also indicate that a reduction in the NADPH cellular pool confers resistance to benzo(a)pyrene. Preliminary data suggest that food restriction may depress G6PDH levels and this may contribute to the tumor preventive effect of underfeeding. PMID- 3024303 TI - [Use of traps in tsetse control: effects of treated and untreated supports]. AB - Tsetse control trials were carried out in forest zone of Cote d'Ivoire with biconical traps. Some of the traps were impregnated with deltamethrin (1, 2 g per trap) and others were not. Results show a rapid and effective action of the treated supports. In nine days, the tsetse population had been reduced by 85% on the treated supports and by 60% on the non treated supports. After one month, a rapid decrease of the efficacy of the treated traps was observed. The fall in the effectiveness (more masked in the treated traps) was certainly due to heavy rains (875 mm) which caused an important increase in growth of vegetation. PMID- 3024304 TI - [Analysis of the dietary fiber level in Polish food products. III. Dietary fiber levels in fruit, vegetables and legume seeds]. PMID- 3024305 TI - [New trends in applying biology to medicine--DNA polymorphism]. PMID- 3024306 TI - Platelets augment granulocyte aggregation and cytotoxicity: uncovering of their effects by improved cell separation techniques using Percoll gradients. AB - The 'standard' technique of granulocyte preparation for in vitro studies uses dextran removal of erythrocytes and Ficoll-Hypaque gradient centrifugation to increase granulocyte purity. The procedure is lengthy, approximately 150 min in our hands, and provides granulocytes significantly contaminated with platelets (approx. 5 platelets/PMN). We report a technique that replaces dextran with hydroxy-ethylstarch and Ficoll-Hypaque with Percoll. Preparation time is reduced by approximately 40% and platelet contamination by more than 80%. Granulocytes, so prepared, function metabolically (O2-generation, chemiluminescence, HMP-shunt maxima) and, in motility/phagocytosis assays, identically to 'standard' preparations. However, an augmentatory effect of platelets in granulocyte aggregation responses and their mediation of cytotoxicity is uncovered. Ficoll Hypaque purified cells (platelet-rich) aggregate to a significantly greater degree with FMLP or activated complement lectins and excessively kill 51Cr labelled target cells when compared to Percoll-preparations (platelet-poor). Re addition of purified platelets or of platelet release supernatants to the latter reproduces results using the 'standard' preparations. PMID- 3024307 TI - Impaired chemiluminescence response by neutrophils in patients with multiple myeloma. AB - Neutrophil chemiluminescence (CL) as a measure of oxygen-dependent killing activity was evaluated in 3 groups of patients: (a) 63 patients with multiple myeloma (MM); (b) 31 subjects with monoclonal gammopathy of undetermined significance (MGUS); (c) 32 healthy controls. Neutrophil CL response was shown to be significant reduced both in patients with MM (p less than 0.001) and in subjects with MGUS (p less than 0.001). A significant difference was also observed between the results obtained in MM and those of MGUS (p less than 0.001). Treated MM patients showed a more severe impairment of neutrophil chemiluminescence response than that observed in untreated patients (p less than 0.001). It is suggested that the impairment of neutrophil CL response, possibly related to decreased killing activity, may play a role, along with other known causes, in the increased susceptibility to infection observed in MM patients. PMID- 3024308 TI - True idiopathic splenomegaly--a distinct clinical entity. AB - 10 asymptomatic young male patients with moderate splenomegaly detected at a routine examination are presented. The history and clinical examination failed to reveal the aetiology of the splenomegaly. Further investigations, including screening for blood dyscrasias, clotting abnormalities and reticuloendothelial abnormalities were likewise unrevealing. Liver biopsies, rectal biopsies for bilharzia and bone marrow aspirates for Gaucher's Disease were found to be normal. Serology for malaria and Ebstein Barr Virus infection was also negative. Positive immunofluorescent tests for IgG antibodies specific for cytomegalovirus were found in 5 patients. We consider that these patients have splenomegaly which is not of a specific nature, but may be associated with a severe antigeneic response to the previous cytomegalovirus infection. In view of the otherwise negative findings these patients should be considered to have 'True Idiopathic Splenomegaly', a term which would indicate the benign nature of the splenic enlargement. This diagnosis should be considered in the differential diagnosis of asymptomatic patients who have splenomegaly of undetermined origin. PMID- 3024309 TI - [Diagnosis, treatment and course of hypophyseal tumors. Retrospective study of 123 cases]. AB - To assess the diagnostic and therapeutic problems associated with pituitary tumors, the clinical records of 123 consecutive patients presenting with radiological changes consistent with pituitary tumors were analyzed retrospectively. This diagnosis was established in 109 cases while the remaining 14 had other intrasellar pathologies. Prolactinoma in women was diagnosed at a significantly younger age (34.3 years) than in men (54.2 years) or than non secreting adenoma in both sexes (52.1 years). In the two latter instances there was, on diagnosis, a larger proportion of visual field impairment due to tumor impingement on the visual pathway. Acromegaly was also recognized late, when skeletal changes were marked. The presence of certain symptoms could have suggested the diagnosis of pituitary tumor earlier in most cases. Surgical treatment was somewhat less effective in correcting hormone hypersecretion than in other series but, on the other hand, it caused less pituitary insufficiency and, thus, less additional discomfort to the patients. Hypopituitarism was frequent after transcranial surgery and radiotherapy (more than 50%) but was very rare after transsphenoidal tumorectomy. It appears that earlier diagnosis would result in less frequent visual impairment and hypopituitarism secondary to tumor extension and the necessary treatment. PMID- 3024310 TI - [Myeloperoxidase deficiency--blemish or disease? Evaluation of 60,337 differential blood pictures]. AB - Leukocyte differential counting by flow cytochemistry (H 6000, Technicon) has shown 32 subjects with partial or complete leukocyte myeloperoxidase deficiency in a population of 60,337 patients screened at a reference laboratory over a one year period. Some of the patients were hospitalized, but most were outpatients. Partial (27 patients) or complete (5 patients) myeloperoxidase deficiency was confirmed by examination of cytochemical stains (Graham-Knoll method). None had hematologic malignancy. Eleven of these 32 patients had recurrent infections; 3 of these patients were found to have a hereditary disturbance which also involved other members of the family. The findings suggest that the incidence of myeloperoxidase deficiency is much higher than was previously suspected. PMID- 3024311 TI - [Fine-needle aspiration biopsy of the breast. Frequency, indication and accuracy, studied on material from the Cytological Laboratory of the Pathology Institute, St. Gallen Canton Hospital, 1981-1984]. AB - 1768 fine needle aspiration biopsies (FNAB) of the breast were evaluated between January 1981 and June 1984 at the Cytologic Laboratory, Institute of Pathology, Cantonal Hospital, St. Gall. The number of FNAB increased from 334 in 1981 to 837 in 1984. By far the most frequent indication for FNAB was a palpable lesion of the breast (in 92.1% of all cases). Other indications were radiological lesions without a palpable mass (2.7%), skin changes of the breast (1.7%) and, in a few cases, unilateral enlargement of the breast. Sensitivity and specificity of the method were calculated on the basis of two groups (A and B). In group A the FNAB was performed by a cytopathologist and in group B by general practitioners and specialists or hospital doctors. Sensitivity of 90.0% was calculated in group A and of 79.0% in group B. The specificity in the evaluated material was 100% for both groups. In group A, 5% of all carcinoma showed a false negative cytologic diagnosis, while in 9% of all FNAB the aspirated material was not representative. Most of the false negative diagnoses occurred with scirrhous carcinoma and with small carcinoma less than 1.5 cm in diameter. The percentage of the cases with a non-representative diagnosis reached 5% of all histologically proven carcinoma in group A and 19.2% in group B. The significantly higher percentage of non representative diagnoses, together with a significantly lower sensitivity in group B (A: 90.0%, B: 79.0%) lead to the conclusion that the experience of the person who has performed the aspiration is of great importance for the diagnostic result.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3024313 TI - [Role of eicosapentaenoic acid in ischemic diseases]. PMID- 3024312 TI - [Pathophysiologic aspects and modern treatment of congestive heart failure]. AB - Congestive heart failure is a clinical syndrome with inadequate cardiac output and increased venous pressure, characterized by hyperfunction of the sympatho neuro-endocrine system, favoring maximal vasoconstriction in non-essential organs and retention of salt and water. Identification of the cause of heart failure and its correction is the best treatment. The suppression of precipitant factors is also essential. Traditionally the treatment of congestive heart failure is based on diuretics, restriction of salt intake, and digitalis. The conventional vasodilators are effective in the short term, but they have a marked tendency to tachyphylaxis. The inhibitors of angiotensin-converting enzyme are, through their specific action on neuro-endocrine dysregulation, the most important advance in recent years. Of the non-glycosidic inotropic agents used in severe heart failure, the new inhibitors of phosphodiesterase need further testing. In certain cases, permanent synchronous pacing has to be considered. PMID- 3024314 TI - [Discovery of GABA]. PMID- 3024315 TI - [Correlations between sonographic and clinical findings in breast cancer]. AB - We recorded the sonographic findings of 108 cases of carcinoma of the breast and compared the different criteria with the respective histological and clinical parameters. When scanned, carcinomas of the breast usually presented as irregular carcinomas with good sound conduction, few internal echoes and rarely with a lateral or dorsal ultrasonic shadow. Sonography showed a smaller diameter of the tumours than measured histologically. Medium-size and solid carcinomas with a smooth surface could be distinguished mos easily. Histological and cytological grading classified these tumours in the high risk group. Tubular and scirrhous tumours were significantly less easy to identify and delineate. This applies also to bigger and smaller tumours. In most cases small carcinomas were smooth, were likely to show sound intensification, and rarely had a dorsal ultrasonic shadow. The bigger they became, the more often we could observe branches; sound intensification decreased, and the dorsal ultrasonic shadow increased. Carcinomas of young women had the same sonographic characteristics as small tumours, whereas carcinomas in elderly women had those of big tumours, although there was no correlation between age and the size of the tumours. We emphasise the importance of sonography in determining tumours dynamics. PMID- 3024316 TI - [Ultrasound-guided incisional biopsy in generalized and focal diseases of the liver]. AB - Sonographically guided liver biopsies under permanent visual control using fine needles are far less traumatic than previous biopsy techniques and carry a very low risk. They may be performed on an outpatient basis, especially when using the newly developed cutting-edge needle which allows to perform core biopsies of various parenchymal organs. Risks are discussed. PMID- 3024317 TI - A deletion truncating the gonadotropin-releasing hormone gene is responsible for hypogonadism in the hpg mouse. AB - Hereditary hypogonadism in the hypogonadal (hpg) mouse is caused by a deletional mutation of at least 33.5 kilobases encompassing the distal half of the gene for the common biosynthetic precursor of gonadotropin-releasing hormone (GnRH) and GnRH-associated peptide (GAP). The partially deleted gene is transcriptionally active as revealed by in situ hybridization histochemistry of hpg hypothalamic tissue sections, but immunocytochemical analysis failed to show the presence of antigen corresponding to any part of the precursor protein. PMID- 3024318 TI - Tissue-specific and ectopic expression of genes introduced into transgenic mice by retroviruses. AB - Recombinant retroviruses containing the complete genomic human beta globin gene (under the control of its own promoter) and the bacterial neomycin phosphotransferase gene (under the control of the normal or enhancerless viral promoter) were used to derive transgenic mouse strains by infection of preimplantation embryos. Expression of the beta globin gene in hematopoietic tissues was observed in all transgenic strains. In addition, one strain showed ectopic expression of beta globin in the same tissues that also expressed high levels of RNA from the viral promoter. It is likely that expression from the long terminal repeat (LTR), in contrast to expression from the internal promoter, is dependent on the site of integration. Thus, retroviral vectors can be used for tissue-specific expression of foreign genes in transgenic mice, as well as for the identification of loci that allow developmental activation of a provirus. PMID- 3024319 TI - How Eco RI recognizes and cuts DNA. PMID- 3024320 TI - The metabolism of phosphoinositide-derived messenger molecules. AB - The phosphoinositides are minor phospholipids present in all eukaryotic cells. They are storage forms for messenger molecules that transmit signals across the cell membrane and evoke responses to extracellular agonists. The phosphoinositides break down to liberate messenger molecules or precursors of messenger molecules. Many different compounds are formed, although the functions of only a few are understood. Recent studies elaborating the pathways for formation of products from phosphoinositides and the factors controlling their metabolism are summarized here. PMID- 3024321 TI - Structure of the DNA-Eco RI endonuclease recognition complex at 3 A resolution. AB - The crystal structure of the complex between Eco RI endonuclease and the cognate oligonucleotide TCGCGAATTCGCG provides a detailed example of the structural basis of sequence-specific DNA-protein interactions. The structure was determined, to 3 A resolution, by the ISIR (iterative single isomorphous replacement) method with a platinum isomorphous derivative. The complex has twofold symmetry. Each subunit of the endonuclease is organized into an alpha/beta domain consisting a five stranded beta sheet, alpha helices, and an extension, called the "arm," which wraps around the DNA. The large beta sheet consists of antiparallel and parallel motifs that form the foundations for the loops and alpha helices responsible for DNA strand scission and sequence-specific recognition, respectively. The DNA cleavage site is located in a cleft that binds the DNA backbone in the vicinity of the scissile bond. Sequence specificity is mediated by 12 hydrogen bonds originating from alpha helical recognition modules. Arg200 forms two hydrogen bonds with guanine while Glu144 and Arg145 form four hydrogen bonds to adjacent adenine residues. These interactions discriminate the Eco RI hexanucleotide GAATTC from all other hexanucleotides because any base substitution would require rupture of at least one of these hydrogen bonds. PMID- 3024322 TI - Proteolytic enzymes in cancer invasion and metastasis. PMID- 3024323 TI - Bolus intramuscular methylprednisolone acetate as a therapeutic adjunct in rheumatoid arthritis. AB - Eighty adult patients with classical or definite rheumatoid arthritis were given 200 bolus injections of intramuscular methylprednisolone acetate (MPA) over four years. This was used strictly as a therapeutic adjunct for polyarticular flares (greater than or equal to 5 swollen joints), not as a singular form of long-term treatment. Good clinical responses, lasting an average of eight weeks, were obtained in the vast majority of patients. Severe toxicity was minimal. Postbolus responses to intravenous ACTH stimulation were measured in 12 patients. We believe that intramuscular bolus injections, used cautiously in carefully selected patients with rheumatoid arthritis, are relatively useful, convenient, inexpensive, and safe. PMID- 3024324 TI - Cytomegalovirus infection in Penang: a serological survey. AB - During 1984-1985, a total of 838 sera obtained from individuals of different age groups, mostly blood donors and those whose sera were received for VDRL tests and other serological investigations. The sera were titrated for complement fixing antibodies against cytomegalovirus (Ad169 strain). Three hundred and fifty two (41%) out of 838 sera showed significant antibody titre. The incidence of this virus infection varied form 26% in the age group of 11-20 years to 59% of those above 50 years of age. Geometric mean titre (GMT) was highest (22) in age groups of 11-20 years and those over 50 years indicating active viral infection in these two age groups. GMT was also significantly higher in females in all age groups except in the age group of 21-30 years and those above 50 years, indicating that active viral infection is more common in females. PMID- 3024325 TI - Aetiology of acute hepatitis in Malaysia. AB - Icteric patients with clinical and biochemical evidence of liver disease, admitted into various hospitals in Malaysia, were investigated to determine the cause of their infection. Of these patients, 11.0% (16/145) were found positive for IgM anti-HAV (EIA), 4.1% (6/145) for IgM anti-HBc (EIA), 1.0% (1/102) for IgM anti-CMV (ELISA), 17.2% (16/64) for rising titres of leptospiral agglutinin, and none for heterophile antibody of EBV. Hepatitis NANB accounted for 67.9% of cases. The mean serum transaminases (ALT and AST) values in patients with hepatitis A and B were higher (more than 500IU) than in patients with leptospirosis or non-A, non-B hepatitis, whereas serum bilirubin levels were higher in patients with hepatitis A and leptospirosis than in patients with hepatitis B. PMID- 3024326 TI - Hepatitis B problem in Thailand. AB - HBV infection is hyperendemic in Thailand. Approximately 5 million Thais are chronic HBV carriers. The prevalence of HBV markers in general population varies from 40-60%. Approximately 10-20% of children between the ages 1-5 years have serologic evidence of HBV infection and this prevalence increases with age reaching a plateau of 40-60% by age 20. High risk groups are household contacts of HBsAg carriers and babies born to HBsAg positive mothers. Approximately 75% of the babies born to HBsAg & HBeAg positive mothers become HBsAg positive at 3 months after birth. A few studies showed that the HBV prevalence of hospital personnel and other high risk groups is similar to that of the general population. The prevalence of chronic HBsAg carrier varies from 5-10% and is highest among age groups 10-30 years. Primary hepatocellular carcinoma (PHC) is the first and third most common cancer among Thai males and females, respectively. Approximately 35%-75% of PHC in adults are HBsAg positive. Histological studies showed that 47.3% of cryptogenic cirrhosis, 58%-66% of PHC and 35%-85% of cryptogenic cirrhosis with PHC were HBsAg positive. Studies on Hepatitis B immune globulin and Hepatitis B vaccine revealed a 70% and 56%, respectively, reduction in the HBsAg prevalence of infants born to HBsAg and HBeAg positive mothers. More epidemiologic, clinical and laboratory studies on HBV infection are being carried out by groups of scientists and investigators in the Ministry of Public Health and many medical schools. A national committee has been appointed to plan strategy for controlling HBV. PMID- 3024327 TI - The National Institute of Mental Health--Epidemiologic Catchment Area (NIMH-ECA) program. Background, preliminary findings and implications. PMID- 3024328 TI - Synovial sarcoma of the thoracic spine. A case report. PMID- 3024329 TI - Pyarthrosis of the sacroiliac joint presenting as lumbar radiculopathy. A case report. PMID- 3024330 TI - Arachnoiditis ossificans of the cauda equina. A case report. PMID- 3024331 TI - Effect of gamma rays at the dihydrofolate reductase locus: deletions and inversions. AB - A series 11 gamma-ray-induced mutants at the dihydrofolate reductase (dhfr) locus in Chinese hamster ovary cells has been examined for the types of DNA sequence change brought about by this form of ionizing radiation. All 11 mutants were found to have suffered major structural changes affecting the dhfr gene. In eight of the mutants, all or part of the dhfr gene has been deleted. The extent of these deletions was examined in seven of these mutants and, for comparison, in two deletion mutants that were induced by UV irradiation. For this purpose, probes from an overlapping set of cosmids that span 210 kb of DNA in this region were used. Three of seven gamma-ray-induced mutants and one UV-induced mutant were shown to have deleted the entire 210-kb region. In the remaining mutants, endpoints ranging from within the dhfr gene to 100 kb downstream were observed. No upstream endpoints were detected, so that an upper limit on the size of these large deletions could not be assigned. Three of the 11 gamma-ray-induced mutants contained an interruption in the dhfr gene without any detectable loss of sequence. Restriction analysis of these interrupted mutants showed that at least 8-14 kb of "foreign" DNA sequence became joined to the gene at the point of disruption. Cytogenetic analysis of these mutants showed that in two cases an inversion of the banding pattern on chromosome Z-2 had taken place. The inverted dhfr mutants contain very low amounts of dhfr RNA sequences, and the 5' end of an inversion mutant gene exhibits the same pattern of DNA methylation and DNase I hypersensitivity as the wild-type gene. Our results suggest that ionizing radiation causes primarily, if not exclusively, large deletions and inversions in mammalian cells. PMID- 3024332 TI - Stability of retrovirally transduced markers in a rat cell line. AB - A MoMLV-based retroviral vector capable of transmitting and expressing both the human hypoxanthine phosphoribosyltransferase (hprt) coding sequence and the Herpes simplex type 1 thymidine kinase (tk) gene has been constructed. After infection of a rat cell line, cell clones were selected on the basis of expressing both markers. They were subsequently found to contain a single provirus of the expected topology. The ease with which loss of expression of the markers can be monitored has allowed us to make observations on the stability of proviral genes. In particular, we have found indirect evidence of strong position effects on proviral gene expression by comparing the characteristic frequency of marker loss in different clonal proviral lines. Effects of the selection protocol on the apparent frequency of variants have also been noted. Finally, a combination of molecular and genetic observations lead us to invoke chromosome loss as the major factor influencing marker stability in this system. PMID- 3024333 TI - Reactivation of X-linked genes in human fibroblasts transformed by origin defective SV40. AB - To determine if expression of genes on the inactive X is inducible in human cells, we looked for reactivation events in a clone of fibroblasts transformed with origin-defective SV40. The karyotype of these cells was grossly heteroploid so that the aneuploidy associated with SV40 transformation occurs even in the absence of viral replication. This transformed clone, heterozygous for hypoxanthine phosphoribosyltransferase (HPRT), lacks HPRT activity, as the mutant allele is on the active X and the normal allele on the inactive X. Reactivation of the HPRT+ allele on the inactive X was observed at a frequency of 6 X 10(-5) per cell and increased approximately eightfold following treatment with the cytidine analogs 5-azacytidine (5azaC) and 5-azadeoxycytidine. The fact that spontaneous reactivation is detectable in some clones, but not all, suggests that the environment of the SV40-transformed cell, although not sufficient to induce generalized derepression, increases the frequency of rare reactivation events. The methylation pattern at the HPRT locus revealed transformation-associated alterations that may have predisposed these cells to reactivation events, spontaneous as well as 5azaC-induced. PMID- 3024334 TI - Detection of amplified sequences in mammalian DNA by in-gel renaturation and SINE hybridization. AB - The presence of amplified sequences in mammalian DNA can be determined by in-gel renaturation of labeled restriction digests of total genomic DNA, provided that such sequences comprise at least 20-30 copies per haploid human or hamster genome or 40-50 copies per mouse genome. To detect amplified DNA in mouse cells at a lower level of amplification, a new procedure has been developed. This procedure combines in-gel DNA renaturation with Southern hybridization using a cloned probe containing a short interspersed repeated element (SINE). Using mouse cell lines containing amplified dihydrofolate reductase (DHFR) or c-Ki-ras genes as a model system, and the cloned B2 repeated sequence as a SINE probe, we have shown that this technique can detect gene amplification at a level as low as 10-15 copies of amplified DNA per haploid mouse genome. In-gel renaturation-SINE hybridization was used to assay for DNA amplification in different tissues and in different mouse strains. No tissue-specific DNA amplification has been detected, but comparison of B2-containing repeated fragments between three inbred mouse lines revealed strain-specific polymorphism. PMID- 3024335 TI - Human insulin-related DNA sequences map to chromosomes 2 and 11. AB - The insulin-related hormone family consists of four known peptides: insulin, the insulin-like growth factors I and II, and relaxin. To investigate the hypothesis that the family contains additional members, we have isolated a series of human genomic clones using the insulin gene as a hybridization probe. Two of these single copy DNA sequences, hIr1 and hIr2, have been localized to chromosome 2 and 11p11----q13, respectively, by Southern blot analysis of mouse-human somatic cell hybrids, a result consistent with other evidence supporting the dispersal of the insulin gene family throughout the genome. Although a biological function for these DNA sequences has not yet been established, hIr1 and hIr2 are potentially useful as molecular probes for mapping, by analysis of restriction fragment length polymorphisms (RFLP), genetic disorders linked to chromosomes 2 or 11. PMID- 3024336 TI - Analgesic actions of naloxone suggest the presence of a hyperalgesic opioid system. PMID- 3024337 TI - Epstein-Barr virus infection in a patient with active pulmonary tuberculosis. A case report. AB - Active pulmonary tuberculosis is highly prevalent among the black population but factors predisposing to a defective cellular immune response to the tubercle bacillus are not readily apparent in many. Serological evidence of a recent reactivation of Epstein-Barr virus (EBV) infection in such a patient could therefore be a relevant finding, since EBV has the potential to suppress cellular immune responses. PMID- 3024338 TI - [Anti-D contaminating human thrombin]. PMID- 3024339 TI - [The amenorrheas]. PMID- 3024340 TI - Preoperative localization and intraoperative glucose monitoring in the management of patients with pancreatic insulinoma. AB - Whipples triad of hypoglycemic episodes associated with fasting blood sugar levels of less than 50 milligrams per 100 milliliters with relief of the symptoms by the administration of glucose intravenously leads to the clinical diagnosis of insulinoma. This diagnosis can be confirmed in a laboratory by the biochemical measurement of an inappropriate insulin elevation in response to fasting or the infusion of calcium or tolbutamide. Since more than 90 per cent of the tumors are benign, the potential for operative cure is high. Unfortunately, because of multicentricity (12 to 13 per cent) and the frequent small size of the tumor, some series report 15 to 30 per cent of inadequate or failed operations. Preoperative localization, in most centers, has depended upon sonography (positive in one of six patients in our series), computerized tomographic scanning (positive in one of five patients), celiac axis angiography (positive in six of 15 patients) and transportal venous sampling for insulin levels (positive in 11 of 13 patients in our series). We found that the combination of arteriography and transportal sampling has been the most accurate means of precise preoperative localization. In conjunction with preoperative localization, we have used intraoperative monitoring of glucose levels as a guide to the completeness of resection of insulin producing tumors. Sustained elevation of blood glucose levels has confirmed the adequacy of surgical intervention. Failure of the blood sugar level to increase had led to the successful search for additional tumors not identified preoperatively or to further resection. The combination of arteriography, transportal sampling and monitoring of glucose levels has led to the cure of 15 patients operated upon at the Mount Sinai Hospital from 1977 to 1984. PMID- 3024341 TI - Local recurrence and the deep resection margin in carcinoma of the breast. AB - Deep resection margin of carcinoma of the breast is an important determinant of the extent of surgical resection as well as the use of adjunctive therapy. To investigate the relationship between deep resection margin and local recurrence, we retrospectively reviewed 105 consecutive female patients with clinical or pathologic Stage II infiltrating ductal carcinoma of the breast. All patients underwent a modified radical mastectomy with the removal of an intact pectoral fascia. The deep margin of resection was determined by histopathologic examination. Detailed follow-up information was obtained for 100 patients. In this study group, the distance from the tumor margin to the pectoral fascia did not correlate with local recurrence. Seventy-seven patients had deep margins of 1 centimeter or less; eight patients had a local recurrence. Recurrence in this group did not correlate with the distance from the deep margin (p = 0.92). The adjunctive administration of radiation therapy or chemotherapy did not appear to influence the frequency of local recurrences. These data suggest a deep margin consisting of an intact pectoral fascia may be adequate to prevent local recurrence in patients with infiltrating carcinoma of the breast. PMID- 3024342 TI - The effect of thyrotropin and cAMP on DNA synthesis and cell growth of human thyrocytes in monolayer culture. AB - Monolayer cultures of human thyrocytes from normal tissue (n = 10), and adenomas (n = 7), differentiated (n = 4), poorly differentiated (n = 2), and undifferentiated (n = 3) thyroid cancers were established to assess the significance of thyrotropin (TSH) and cAMP (adenosine 3',5'-cyclic monophosphate) on cell growth and DNA (deoxyribonucleic acid) synthesis. Cell growth of thyrocytes from normal and adenomatous tissues increased more rapidly (p less than 0.01) after TSH (0.1 IU/ml) was added but was unaffected by cAMP (10(-4) mol/L). In these cells, TSH also enhanced DNA synthesis twofold to twelvefold (p less than 0.01). The adenylate cyclase (AC) inhibitor, 2',3' dideoxyadenosine (ddA), increased DNA synthesis 1.3 to 6 times at a concentration of 2 X 10(-4) mol, whereas the membrane/passable cAMP analogue, dibutyryl-cAMP, and the AC stimulator, forskolin, failed to show any effect on DNA synthesis up to a concentration of 10(-5) mol/L (p less than NS). When administered simultaneously, TSH (1/2 maximum) and ddA (20 mumol) had no cumulative effect on DNA synthesis (p = NS). TSH stimulation in cancerous thyroid tissue (n = 11) demonstrated a lack of TSH response in seven of 11 monolayer cultures with no apparent correlation to cancer differentiation, patient age, or sex. Thus TSH was demonstrated to stimulate DNA synthesis and cell growth of human thyrocytes in monolayer cultures independent of the AC system. However, the TSH effect on cell growth and DNA synthesis was unpredictable in thyrocytes from cancerous tissues. PMID- 3024344 TI - [A case of pleomorphic adenoma of the mouth]. PMID- 3024343 TI - Nonfunctioning malignant neuroendocrine tumors of the pancreas. AB - The clinical features of eight women and three men with nonfunctioning islet cell carcinoma of the pancreas were reviewed. The mean patient age was 58 years (range 44 to 75 years). Weight loss and abdominal pain were the most frequent presenting symptoms. An abdominal mass was palpable in five patients. At operation regional or distant metastases were present in 82% of patients. Only 18% of patients underwent resection for potential cure. All tumors proved histologically to be neuroendocrine in origin. Immunohistochemical staining showed positive reactivity for neuron-specific enolase and chromogranin in all tumors studied but was negative for insulin, glucagon, and somatostatin. Focal positivity for pancreatic polypeptide was seen in one tumor. Nine patients with unresectable disease at operation were available for follow-up. Mean survival for the entire group was 23 +/- 7.2 months (range 4 to 72 months). Survival differences between women and men appeared to favor women but were not statistically significant. Postoperative regional or systemic chemotherapy also had no significant effect on patient survival although two of the longest survivors (36 and 72 months) had received adjunctive chemotherapy. Nonfunctioning islet cell neoplasms are locally aggressive, have a propensity for early metastases, and are rarely resectable for cure. Unlike pancreatic exocrine carcinomas, endocrine malignancies may respond favorably to adjunctive chemotherapy. PMID- 3024345 TI - [New aspects in the diagnosis and treatment of herpes infections]. PMID- 3024346 TI - Resuscitative laser photoresection of a tracheal tumour before elective surgery. PMID- 3024348 TI - Bronchial brushing and bronchial biopsy: comparison of diagnostic accuracy and cell typing reliability in lung cancer. AB - A total of 443 patients with lung cancer underwent brush and forceps biopsy through a fibreoptic bronchoscope. The biopsy was taken from the area of suspected malignancy which had been brushed. Of 443 patients, 400 (90.3%) showed positive results on brushing and 287 (64.8%) on biopsy. A combination of both techniques yielded the highest percentage of positive diagnosis (93.7%). Histologically, there was a high incidence of positive diagnosis for squamous and small cell carcinoma. One hundred and three (83.7%) of 123 specimens obtained by brushing and 75 (81.5%) of 92 specimens obtained by biopsy agreed with the cell type found in the surgical or necropsy specimen. Cell typing accuracy was higher in squamous and in small cell carcinoma in both techniques. As the cell typing accuracy of the two methods is similar, the results obtained by both techniques should be taken into consideration in the management of individual cases of lung cancer. PMID- 3024347 TI - Lung collagen: more than scaffolding. PMID- 3024349 TI - Survival in small cell lung carcinoma after surgery. AB - In a retrospective study of long term survival in patients with small cell carcinoma of the lung who had been treated purely by surgery, 1820 patients with lung cancer seen during the 15 years 1962-77 were reviewed and reclassified histologically and according to the TNM system. Of these patients, 924 had had resections and 284 exploratory thoracotomies. Cancer chemotherapy was not used in this period and radiotherapy was given only occasionally as palliative treatment. Seventy seven of the patients having pulmonary resections had small cell carcinoma (8.4%), and there were six survivors among the 71 with T1-2, N0-1, M0 tumours. The five and 10 year survival rates were both 12%. The histological specimens from these six patients with a small cell carcinoma who survived more than 10 years were re-evaluated and confirmed as small cell by an independent group of pathologists. It seems justified to conclude that a selected group of patients with small cell carcinoma should be treated by surgery alone without adjuvant chemotherapy, which might reduce the long term survival. PMID- 3024350 TI - Relationship between changed alveolar-capillary permeability and angiotensin converting enzyme activity in serum in sarcoidosis. AB - The effect of altered alveolar-capillary permeability on angiotensin converting enzyme (ACE) activity in serum (SACE) was studied in 45 patients with sarcoidosis and 21 healthy controls. In sarcoidosis increased albumin concentrations in the bronchoalveolar lavage fluid (L albumin) and increased ratios of L albumin to albumin in serum (S albumin) indicated an increased permeability of the alveolar capillary membrane. ACE activity in the lavage fluid (LACE) was correlated with the number of alveolar macrophages in controls, indicating that it may come from these cells. LACE was high in active sarcoidosis while in inactive disease it was similar to that in controls. SACE in sarcoidosis was significantly increased. Ninety per cent of patients with increased L albumin had increased SACE. SACE activity was significantly correlated with concentrations of L albumin and with LACE activity. The relationships between signs of increased membrane permeability and SACE and between LACE and SACE suggest that excess SACE in sarcoidosis may, at least partly, originate in the alveolar space. PMID- 3024351 TI - Short duration combination chemotherapy in the treatment of small cell lung cancer. AB - Ninety five patients (57 with limited disease and 38 with extensive disease) with previously untreated small cell lung cancer were entered into a study of short duration combination chemotherapy with intravenous cyclophosphamide (750 mg/m2) on day 1, adriamycin (40 mg/m2) on day 1, and etoposide VP-16 (100 mg/m2) on days 1, 2, and 3, with the addition on day 10 of methotrexate 50 mg/m2 with folinic acid rescue and vincristine 2 mg. The treatment was repeated on day 22 and only three courses were given. No maintenance chemotherapy was given, though patients with a complete response received radiotherapy (30-40 Gy (3000-4000 rads] to the primary site in most cases. Forty nine patients (86%) with limited disease achieved a response, with 26 (46%) complete remissions. Twenty five patients (66%) with extensive disease had a response, but only eight (21%) had a complete response. Actuarial survival analysis for the whole patient population showed a median survival of 13 months for patients with limited disease and seven months for those with extensive disease. The median survival was 14 months for those patients with limited disease who achieved a complete response, but only 10 months for non-responders. Myelosuppression was the major expression of toxicity. There were three deaths related to treatment and seven patients had febrile episodes during neutropenia that required antibiotics. Mucositis, which was usually mild, occurred in 49% of patients. The primary site was the main site of initial relapse in 56% of the patients who relapsed. Among patients with limited disease who achieved a complete response, relapses at the primary site were less common in those who received radiotherapy (five out of 12) than in those who did not (all eight). The results indicate that this short duration chemotherapy in small cell lung cancer gives response rates and the potential for long term survival similar to those obtained in other series while allowing patients the maximum time free from treatment. PMID- 3024352 TI - Increased thromboxane A2 generation and altered membrane fatty acid composition in platelets from patients with active angina pectoris. AB - Lipid composition of platelet membranes and thromboxane A2 (TxA2) generation by platelets were investigated in eighty-seven anginal patients (forty-two with resting angina in active phase and forty-five with effort stable angina or rest angina in inactive phase) and in forty-five clinically healthy subjects of similar age. All subjects were on the same dietary regimen and the adherence to diet was checked by analysis of red blood cell lipids. Platelets from active angina patients produced more TxA2 than platelets from both inactive patients and controls (p less than 0.001). Moreover patients with active angina had higher arachidonic acid (AA, p less than 0.001) and lower eicosapentaenoic (EPA) and docosahexaenoic acid (DHA) levels in phosphatidylcholine (PC, p less than 0.001), than inactive patients and controls. AA and EPA changes in membrane PC significantly correlated with TxA2 production (p less than 0.001) but not with coronary pathoanatomy. Plasma lipids, content of cholesterol, total phospholipids (and their saturated and unsaturated fatty acids) and the different phospholipid fractions in platelet membrane were not different in the three groups. Present results indicate that in platelets from anginal patients phospholipid fatty acid composition is at least in part independent of plasma composition and that in active angina there are modifications leading to increased TxA2 formation and possibly contributing to the occurrence of ischemic attacks. PMID- 3024353 TI - A comparison of methods of removing inhibitors to the chromogenic Limulus assay in normal and jaundiced blood. AB - A comparative study was performed of methods of removing inhibitors to the quantitative chromogenic method of Limulus assay for endotoxin in normal and jaundiced plasma. Chloroform treatment failed to remove inhibitors in normal and jaundiced plasma and did not prevent inhibition of colorimetry by bilirubin. Perchloracetic acid (PCA) treatment decolourised jaundiced samples but gave unreliable results due to poor pH control. Boiling and dilution gave reliable results in jaundiced and normal plasma without significant loss of sensitivity and is the method of choice. PMID- 3024355 TI - [Paramedical treatment in Trondelag. A study of patients with eye diseases]. PMID- 3024354 TI - Phosphorylation of phosphoinositides in human platelets. AB - 32P-labelling of phosphatidylinositol (PI), PI-4-monophosphate (PIP), PI-4,5 bisphosphate (PIP2) and phosphatidic acid (PA) in 32P-labelled intact human platelets was investigated in the presence of various agents which alter intracellular level of cAMP or Ca2+. Addition of dibutyryl cAMP to intact platelets pre- or pulse labelled with 32P resulted in increased 32P-labelling of PIP and in concomitant decreased 32P-labelling of PI without affecting that of PIP2 or PA. Similar changes were observed in intact platelets treated by prostaglandin I2 (PGI2) or a new low Km phosphodiesterase inhibitor (DN-9693). When intracellular Ca2+ was chelated by loading quin 2-AM to intact platelets, 32P-labelling of PIP was significantly increased in a dose related manner. From these observations it was concluded that PI kinase is activated by elevation of cAMP or chelation of Ca2+ in intact platelets. PMID- 3024356 TI - Potential role of activated macrophages in acetaminophen hepatotoxicity. I. Isolation and characterization of activated macrophages from rat liver. AB - Twenty-four hours following treatment of rats with the analgesic acetaminophen (1.2 g/kg), we observed an infiltration of mononuclear cells into centrilobular regions of the liver in the absence of necrosis. To determine whether acetaminophen induces the accumulation and activation of mononuclear phagocytes, we compared the morphologic and functional characteristics of macrophages obtained from livers of acetaminophen-treated rats with those of resident macrophages (Kupffer cells) from untreated control animals. Macrophages were isolated from rat livers by combined collagenase/pronase perfusion, selective digestion, and differential centrifugation on a metrizamide gradient. Acetaminophen treatment resulted in a twofold increase in macrophage yields from the liver compared with controls. Macrophages isolated from treated animals were generally larger than resident Kupffer cells, were highly vacuolated, and adhered to culture dishes more rapidly. Liver macrophages from both treated and untreated rats phagocytized sheep red blood cells (sRBC) in a time-dependent manner, reaching a maximum after 60-75 min incubation with sRBCs. However, macrophages from livers of acetaminophen-treated rats phagocytized two to three times more sRBC than did resident Kupffer cells. Using the Boyden chamber technique, both macrophage populations were found to be chemotactic to a number of stimuli including the complement fragment, C5a, and synthetic collagenous peptides related to tissue breakdown products. Levels of migration of macrophages from livers of acetaminophen-treated rats were four to seven times greater than those of resident Kupffer cells. In addition, compared with resident Kupffer cells, macrophages from acetaminophen-treated rats released 30% more superoxide anion in response to the stimulus, 12-O-tetradecanoyl-phorbol-13-acetate. Taken together, these results suggest that acetaminophen treatment of rats leads to the recruitment and activation of macrophages in the liver. PMID- 3024357 TI - Potential role of activated macrophages in acetaminophen hepatotoxicity. II. Mechanism of macrophage accumulation and activation. AB - Treatment of rats with acetaminophen (1.2 g/kg) results in the accumulation of activated macrophages in the centrilobular regions of the liver. To study the mechanism by which these cells accumulate and become activated, we examined the release of chemotactic and activating factors from cultured hepatocytes treated with acetaminophen (10-100 microM). We found a dose- and time-related generation of Kupffer cell and monocyte chemotactic activity by acetaminophen-treated hepatocytes. The maximum response was observed with a 25% dilution of medium collected 24 hr following treatment of hepatocytes with acetaminophen. Using a checkerboard assay, the factor in conditioned medium was determined to induce chemotaxis as well as chemokinesis in both Kupffer cells and monocytes. The hepatocyte-derived chemotactic factor was also found to be stable to freeze thawing but to lose activity following heat or trypsin treatment. These results, together with our findings that chemotactic activity was eluted in the void volume following Sephadex G-25 size exclusion chromatography, suggested that the chemotactic factor released by hepatocytes is a large molecular weight protein. The release of Kupffer cell activating factors by acetaminophen-treated hepatocytes was also examined. Hepatocyte-conditioned medium was found to stimulate Kupffer cell phagocytosis and superoxide anion release, two characteristics of activated macrophages. These effects were maximal with conditioned medium collected from hepatocytes 24 hr following treatment with 50 100 microM acetaminophen. Acetaminophen alone had no effect on chemotaxis, phagocytosis, or superoxide anion production by Kupffer cells or monocytes. Taken together, these results suggest that macrophage accumulation and activation in the liver following acetaminophen treatment may be mediated, at least in part, by factors released from hepatocytes. PMID- 3024358 TI - Ah receptor mediated suppression of the antibody response in mice is primarily dependent on the Ah phenotype of lymphoid tissue. AB - Halogenated aromatic hydrocarbons act through the aromatic hydrocarbon (Ah) receptor in mice to produce a series of toxic effects of the immune system. The receptor protein is a product of the Ah gene locus. Ah responsive (Ahb/Ahb) mice express a high affinity receptor in both lymphoid and nonlymphoid tissues whereas nonresponsive Ahd/Ahd mice express a poor affinity receptor. To determine the role of the Ah receptor of lymphoid tissue relative to that of nonlymphoid tissue in the induction of immune impairment, bone marrow was used to reconstitute lethally irradiated mice of the same or opposite Ah phenotype. All mice were given 3,3',4,4'-tetrachlorobiphenyl (35 and 350 mumol/kg) ip 2 days before immunization with sheep erythrocytes (SRBC). The immune response to this T dependent antigen and organ weights were determined 5 or 7 days later in normal or chimeric mice, respectively. Monoclonal Lyt 1.1 and Lyt 1.2 antibodies were used to establish the origin of the cells which repopulated the chimeric thymuses. The immune responses of both BALB/cBy (Ahb/Ahb) and the BALB/cBy X DBA/2 hybrid, CByD2F1 (Ahb/Ahd), were significantly suppressed but DBA/2 mice were unaffected. The immune responses of chimeric BALB/cBy----BALB/cBy and BALB/cBy----DBA/2 (donor----recipient) mice were also significantly suppressed and thymic atrophy was observed in both cases. The serum anti-SRBC antibody titers of DBA/2----BALB/cBy chimeras were also significantly decreased although not to the same extent as in BALB/cBy----DBA/2 mice. Chimeric DBA/2----DBA/2 mice were not affected. These results indicate that the sensitivity to Ah receptor mediated suppression of the antibody response is primarily determined by the Ah phenotype of the lymphoid tissue. PMID- 3024359 TI - Nicotine potentiates superoxide anion generation by human neutrophils. AB - Cytotoxic neutrophil-derived oxygen radicals have been implicated in the pathogenesis of a variety of cardiovascular, pulmonary, and neoplastic disorders for which cigarette smoking is a prominent risk factor. Although nicotine alone failed to provoke neutrophil oxidative metabolism, the alkaloid caused dose dependent (0.1 to 10 microM) potentiation of superoxide anion release induced by either phorbol myristate acetate or N-formyl-methionyl-leucyl-phenylalanine. The potentiating effect of nicotine was not attenuated by either atropine or hexamethonium nor was it mimicked by acetylcholine, suggesting involvement of noncholinergic receptors or a membrane-fluidizing effect of the alkaloid. Nicotine-induced exacerbation of neutrophil superoxide anion production may be involved with the enhanced risk of cardiovascular, pulmonary, or neoplastic disease in individuals who smoke. PMID- 3024360 TI - The accumulation of pyrophosphate by rat hepatocytes. AB - Hepatocytes that were isolated from 48 hr starved rats and incubated in Krebs Henseleit bicarbonate buffer containing 10 microM A23187, 10mM l-lactate, 1mM pyruvate and 2mM l-lysine were found to contain 0.064 mumol of inorganic pyrophosphate/g wet wgt cells. Addition of either 20mM acetate or butyrate, which caused the formation of pyrophosphate in both the cytosol and the mitochondrial matrix or only the matrix, resulted in an increase of 0.915 and 1.91 mumol pyrophosphate/g wet wgt cells, respectively. The accumulation of pyrophosphate was shown to be non-linear with time and dependent on the calcium concentration of the incubation media. In contrast, incubations containing a combination of 10 mM NH4Cl and 5 mM ornithine, which resulted in the formation of pyrophosphate only in the cytosol, had a pyrophosphate content of 0.032 mumol/g wgt cells. When isolated hepatocytes that had been incubated with acetate or butyrate were subjected to digitonin fractionation, all of the recoverable pyrophosphate was present in the particulate fraction. It is concluded that pyrophosphate accumulates in isolated rat hepatocytes only in the presence of calcium and a calcium ionophore, only within the mitochondrial matrix and only when pyrophosphate is formed within the mitochondrial matrix. PMID- 3024361 TI - The cytosolic receptor binding affinities and AHH induction potencies of 29 polynuclear aromatic hydrocarbons. AB - The dose-response rat hepatic cytosolic receptor-binding avidities, aryl hydrocarbon hydroxylase (AHH) and ethoxyresorufin O-deethylase (EROD) induction potencies in rat hepatoma H-4-II E cells in culture were determined for 29 polynuclear aromatic hydrocarbons. It was apparent that the magnitude of the EC50 values for these in vitro responses were strongly dependent on structure. Dibenz[a,h]anthracene (1.6 X 10(-8) M), 7-methylbenz[a]anthracene (1.6 X 10(-8) M), 3-methylcholanthrene (2.8 X 10(-8) M) and picene (4.5 X 10(-8) m) exhibited the highest affinity for the receptor protein and these compounds were only 5 fold less active the 2,3,7,8-tetrachlorodibenzo-p-dioxin (1 X 10(-8) M). All of the compounds which were active in the receptor-binding and monooxygenase enzyme induction assays possessed one common structural feature, namely the presence of a phenanthrene structure fused with at least 1 benzo ring. The results also demonstrated that there was not any apparent correlation between the receptor binding avidities and in vitro monooxygenase enzyme-induction potencies for the most active polynuclear aromatic hydrocarbons. PMID- 3024362 TI - Further studies on the formation of oxygen radicals by potassium superoxide in aqueous medium for biochemical investigations. AB - Potassium superoxide (KO2) forms superoxide anion (O2-) in aqueous medium as measured by the superoxide dismutase (SOD)-inhibitable cytochrome c reduction assay of McCord and Fridovich. The reduction of cytochrome c by O2- formed by KO2 was observed only above pH 7.0 and demonstrated a pH optimum at pH 9.6. SOD was an effective inhibitor of the cytochrome c reduction produced by KO2. Hydroxyl radical scavengers and singlet oxygen quenchers did not interfere with the reduction of cytochrome c by KO2. These data demonstrate that addition of KO2 to aqueous medium is an easy chemical method for the production of O2- in controlled amounts. PMID- 3024363 TI - [Augmentation of the residual alveolar ridge of the mandible]. PMID- 3024364 TI - Experimental assessment of the vector competence of Culex (Culex) neavei Theobald with West Nile and Sindbis viruses in South Africa. AB - Culex univittatus is the maintenance vector of West Nile (WN) and Sindbis (SIN) viruses on the inland plateau of South Africa and also transfers infection to man. Both viruses have frequently been isolated from this mosquito and human immunity is high on the plateau. In the Natal lowlands where Cx univittatus is replaced by Cx neavei, apparently the main vector there, immune rates are low and few isolations have been made from this species. Vector competence experiments were carried out with Cx neavei to compare with those previously done with Cx univittatus. It was thought that a difference in such competence might explain the contrasting epidemiological situation in the two regions. After infective feeds on chicks with high viraemias of each virus, Cx neavei showed a high transmission rate. However, the 50% infection thresholds determined by exposing birds with differing viraemias to mosquitoes of this species, were 4.4 logs per ml (WN) and 5.8 logs (SIN). These thresholds are both higher than those for Cx univittatus. It is concluded that Cx neavei is a poorer vector of both viruses which accounts for the lower viral activity, including the lower incidence of human infection, in the Natal lowlands. PMID- 3024365 TI - Onchocerca volvulus: enzyme polymorphism in relation to the differentiation of forest and savannah strains of this parasite. AB - Isozyme analysis was carried out on Onchocerca volvulus worms collected from Liberia, Ivory Coast, Burkina Faso and Sudan to see whether this technique could detect differences between forest and savannah populations of this parasite. A total of 243 forest and 189 savannah individual female worms were electrophoresed and stained for seven enzymes. Four showed some polymorphism, LDH, MDH, PGM and MPI and the other three, GAPDH, PEP and GPI were invariant. Statistical analysis of the results showed that the relative proportions of genotypes from within the different countries conformed to Hardy-Weinberg expectations. Pairwise comparisons of allele frequencies between countries showed that populations from Liberia and Ivory Coast had a very similar composition; there was some divergence between all the other pairs of populations and the genetic distance was calculated to summarize the degree of divergence. The number of loci examined was small and the genetic distances were within the range expected for separate geographical populations of the same species. The usefulness of this technique in worm identification is discussed. PMID- 3024366 TI - Neutralizing antibody response to Japanese encephalitis inactivated mouse brain vaccine among laboratory personnel. AB - Among vaccinees given two doses of JE Nakayama strain liquid vaccine, 41.8% had significant neutralizing (N) antibody response to the P20778 strain and 48.6% to the Japanese Nakayama strain. Among vaccinees who received three doses of the freeze-dried vaccine, the proportion of positive reactors was 53.8% when the sera were tested with the P20778 strain and 47.4% with the Nakayama strain. A fairly large proportion of those vaccinees who had responded positively to the latter vaccine were found to have lost detectable N antibodies to the P20778 strain and a smaller proportion to the Nakayama strain 13 to 17 months after the third dose of vaccine. Following the administration of a booster dose of the vaccine after this time interval, 65.2% and 56.5% of vaccinees responded positively to the P20778 strain and the Nakayama strain, respectively. The differences between the proportion of positive reactors to the two strains were not significant either for the liquid vaccine or the freeze-dried vaccine. An important finding was the priming effect of infection with West Nile (WN) virus before vaccination. Those vaccinees who had N antibody to WN virus before vaccination had a significantly higher N antibody response to the P20778 strain of JE virus than those who had no detectable antibody to WN virus. These findings indicate that the JE Nakayama strain vaccine would be efficacious in India, particularly in view of the widespread prevalence of WN virus infection. PMID- 3024367 TI - Malaria and ELISA HTLV-III antibody reactivity. PMID- 3024368 TI - Expression of HLA-DR antigens on peripheral blood T lymphocytes and renal graft tubular epithelial cells in association with rejection. AB - Since the expression of HLA-DR antigens on peripheral blood T lymphocytes and renal graft tubular epithelial cells may be associated with immunological stimulation, we investigated the expression of these antigens on blood T lymphocytes and graft tubular epithelium in 84 renal transplant patients. Peripheral blood T lymphocytes were monitored by flow cytometry during the first 3 months after transplantation. Since the DR+ lymphocytes were selected by a double-labeling technique used for the Leu2a phenotype, the majority of the DR+ lymphocytes were also Leu2a+ with a small percentage of unidentified Leu2a- lymphocytes. Forty-one patients were treated with cyclosporine (CsA) and 43 with azathioprine (Aza), while both groups received low-dose steroids. Frozen sections of 57 renal biopsies from 43 patients (21 on CsA and 22 on Aza) were stained for HLA-DR antigens. In the Aza group, clinical rejection episodes correlated with an increased percentage of DR+ peripheral lymphocytes (P = 0.0005), and the expression of DR antigens on graft epithelial cells (P less than 0.001). In the CsA group, no relation between the expression of DR antigens on blood lymphocytes and clinical rejection episodes was evident, and the correlation between tubular DR staining and clinical rejection episodes was weaker than in the Aza group (P = 0.03). In both the Aza and CsA group, an increase in DR+ peripheral lymphocytes correlated with positive staining of the renal tubular cells for HLA-DR antigens (P less than 0.001). PMID- 3024369 TI - Pulmonary complications of orthotopic liver transplantation. AB - Pulmonary complications following orthotopic liver transplantation (OLT) were prospectively evaluated in 18 individuals transplanted at the New England Deaconess Hospital. Of sixteen patients who survived the immediate postoperative period, 12 (75%) sustained a pulmonary complication. Of these complications, 64% were noninfectious--whereas 22% were infectious, and 14% probably infectious. Six of eight documented infections were caused by viruses of the herpes group. In four cases of viral pneumonitis other pulmonary pathogens were isolated (fungi-3, protozoan-1, bacteria-1). Unlike noninfectious complications, pulmonary infections were associated with a fatal outcome in five of six patients who died after OLT. Pulmonary complications are frequent and serious occurrences after OLT, and contribute to both the morbidity and mortality of this procedure. Compared with pulmonary complications seen after transplantation of other organs, OLT was associated with a higher proportion of noninfectious complications but a similar spectrum of pulmonary infections. PMID- 3024370 TI - Interstitial pneumonitis following autologous bone marrow transplantation. AB - The incidence of interstitial pneumonitis (IP) was reviewed in 70 consecutive patients who received autologous marrow transplants for hematologic malignancies. All patients were treated with total-body irradiation (TBI), with or without other chemotherapeutic agents. Seven patients (10%) developed IP, 3 were due to cytomegalovirus, 1 due to Pneumomcystis carinii, and 3 of unknown cause (idiopathic). Risk factors for developing IP were increasing age and a prior history of irradiation to the chest. The use of methotrexate posttransplant did not increase the incidence of IP. PMID- 3024371 TI - A prospective study of unexplained nausea and vomiting after marrow transplantation. AB - We prospectively studied patients with enigmatic nausea and vomiting after allogeneic marrow transplantation to define the causes of this syndrome. Fifty consecutive episodes of persistent vomiting were investigated using physical examination and laboratory tests, endoscopic biopsies and brushings, and clinical follow-up for four weeks. Potential causes of vomiting were identified in 39 of the 50 cases (78%). Fifteen cases had gastrointestinal infections (mainly herpesviruses), 13 had unsuspected acute intestinal graft-versus-host disease (GVHD), 8 had intestinal infection plus acute GVHD, and 3 had other causes (subdural hematomas, bacteremia, and encephalitis). In the remaining 11 cases, no cause of vomiting was found. Endoscopy was necessary for diagnosis in 36 cases and required a combination of methods: routine histology, cytology, viral culture, and immunohistology using monoclonal antibodies to cytomegalovirus (CMV) and herpes simplex virus type 1. Patients with unexplained vomiting or intestinal GVHD had significant improvement of nausea and vomiting over the four-week observation period, but those with CMV did not (P = .01). We conclude that most allogeneic marrow transplant patients with enigmatic nausea and vomiting have gastrointestinal herpesvirus infections, acute GVHD, or both. Untreated CMV infections and persistent GVHD are associated with protracted vomiting in these patients. PMID- 3024372 TI - Isolation of rat pancreatic islets by ductal injection of collagenase. PMID- 3024373 TI - [Cancer of the cervix. I. A disease transmitted by venereal infection?]. PMID- 3024374 TI - [Polyneuropathy associated with monoclonal serum proteins and plasma cell dyscrasias]. PMID- 3024375 TI - Percutaneous renal biopsies: accuracy, safety, and indications. AB - Fifty-one percutaneous renal needle biopsies were performed on 46 patients. Initial biopsy was adequate for diagnosis in 89% of patients. When a second biopsy was performed, this accuracy increased to 98%. Thirty-four of 51 (67%) biopsies were for renal masses and 17 (33%) for medical indications. Computed tomographic guidance was utilized in 94% of cases. Biopsies of renal masses were performed with 18-21-gauge needles, while biopsies for medical indications were performed with an 18-gauge cutting needle or 14-gauge Trucut. A rate of serious complications of 6% was noted. PMID- 3024376 TI - [Studies of the cell kinetics of human malignant testicular tumors]. AB - The cellular DNA-content of 75 malignant testicular germ-cell tumours was determined by flow cytometry. The results are: Aneuploidy is a certain tumour marker in 95% of germ-cell tumours. The DNA-indices range from 2 c (diploid) to 6 c with a modal value around 3 c. 23% of germ-cell tumours are multiclonal. The S phase percentage is higher than those of the most other solid tumours investigated so far and is 25.5% in seminomas and 35% in non-seminomatous germ cell tumours. Further investigations of a greater number of tissues will show the importance of DNA-contents measurement by flow cytometry for the prognosis of testicular tumours, representing an addition to histopathology. PMID- 3024377 TI - Oncogenic viruses of domestic animals. AB - The objective of this article is to review the relevant oncogenic viruses of small animals. Basic scientific principles that have been discovered by research into feline leukemia and other animal cancer viruses have enhanced our understanding of oncogenesis and have led to practical methods of cancer control and prophylaxis. PMID- 3024379 TI - Parainfluenza virus type 3 in pigs. PMID- 3024378 TI - Monoclonal antibodies. Clinical uses and potential. AB - An overview of monoclonal antibody technology and some examples of its relevance to veterinary medicine are presented in this article. A technical description of the generation of immune spleen cells and hybridization is included. Feline leukemia, canine parvovirus, and their respective diseases are included as examples of cases in which monoclonal antibodies can be applied in the diagnosis and characterization of these diseases and their etiologic agent. PMID- 3024380 TI - Influenza A virus infection of a pheasant. PMID- 3024381 TI - Bovine herpes mammillitis therapy. PMID- 3024382 TI - Enzyme-linked immunosorbent assay for single serum diagnosis of canine parvovirus disease. PMID- 3024383 TI - Studies on kinetic properties of non-specific phosphomonoesterases in the sheep nematode, Bunostomum trigonocephalum Rudolphi, 1808. AB - The maximum activity (Vmax) of acid phosphomonoesterase (E.C.3.1.3.2.) at pH 5.5 and 37 degrees C was found to be 2.68 +/- 0.25 and 3.85 +/- 0.24 mu moles phenol mg protein-1 min-1 in male and female Bunostomum trigonocephalum, respectively. The Vmax of alkaline phosphomonoesterase (E.C.3.1.3.1) at pH 10.0 and 37 degrees C was 0.75 +/- 0.04 and 1.15 +/- 0.05 mu moles phenol mg protein-1 min-1 in male and female B. trigonocephalum, respectively. The Michaelis constant (Km) values were 10.25 mM and 11.76 mM for acid and 8.69 mM and 9.09 mM for alkaline phosphomonoesterase in male and female worms, respectively. Enzymal activities were optimum at 7.0 and 9.0% enzyme concentrations, at incubation periods of 60 and 20 min and at temperatures of 50 and 45 degrees C for acid and alkaline phosphomonoesterases, respectively. Dialysis in distilled water decreased the activity of both enzymes, while only acid phosphomonoesterase activity increased in citrate buffer (pH 5.5) and alkaline phosphomonoesterase activity in carbonate buffer (pH 10.0). PMID- 3024384 TI - Human effects of veterinary biological products. AB - A 14-year-old male drank two glasses of milk from a gallon inoculated with 21 vials of live virus vaccine intended to immunize 1000 baby chicks against Newcastle Disease. The patient was managed with catharsis and careful observation, and remained completely asymptomatic in the following 28 days. Newcastle Disease virus (NDV) may cause paralytic neurotropic, viscerotropic, or pneumonotropic disease in chickens, but has limited effects on humans. Systemic exposure to this vaccine has not previously been reported in humans, although an aerosol NDV vaccine has caused self-limited conjunctivitis following ocular exposure. The human effects of veterinary biologicals remain poorly defined. The limited literature existing in regards to the human experience with common veterinary vaccines is reviewed. PMID- 3024385 TI - Effect of pre and postnatal lithium chloride ingestion on the developing mouse. AB - The effect of pre and postnatal maternal ingestion of 1 mEq LiCl, given ad libitum in drinking water, on neonatal development was studied in the mouse. The measurements made included evaluation of body and selected organ weights of the weaning offspring. In addition, determination of weaning mouse liver alcohol dehydrogenase (L-ADH), aldehyde dehydrogenase (L-ALDH) were performed to assess the sensitivity of the offspring to ethanol intoxication due to the clinical trials of lithium salts in alcoholism. Heart lactate dehydrogenase isoenzymes were also assayed to assess developmental aspects of the offspring. Little changes occurred in body weight and only moderate increase in spleen weight was noted without concomitant changes in brain, kidney or liver weight of the nursing dams. Maternal Li+ exposure resulted in increased kidney, liver and spleen but not brain weights from controls in the weaning animals. This effect was greater in the female than in the male offspring. The weaning mice showed an induction of L-ALDH as a consequence of maternal Li-treatment. No changes occurred in the other hepatic and cardiac enzymes studied. The results suggest sex-dependent neonatal sensitivity towards maternal ingestion of a small concentration of LiCl in drinking fluid. Hepatic changes of L-ALDH may represent an early expression of hepatic toxicity in the weaning mouse. PMID- 3024386 TI - Host cell modulation of foot-and-mouth disease virus procapsid synthesis. AB - BHK21 (clone 13S) cells of high (BHK-SH) and low (BHK-SL) passage number were infected with foot-and-mouth disease virus (FMDV) subtypes A24, A25 and C3. While the amount of virus specific RNA produced in BHK-SH cells was 25% of that in BHK SL cells and the virion production was 27% (C3) to 53% (A24) lower, the synthesis of viral proteins was comparable, associated with an accumulation of procapsids in BHK-SH cells. The results suggest that changes in viral infection pattern with increasing BHK21 cell passage number should be considered in FMDV vaccine production. PMID- 3024387 TI - [Structure and mechanism of action of restriction endonucleases]. AB - The structure and the mode of action of restrictional endonucleases are dealt with in detail. Described are some more important representatives of the three types of endonucleases. Stated are their role and place in present-day molecular biology. PMID- 3024388 TI - [Isolation of lymphocyte cultures from leukemic cattle and electron microscopic study of the leukemia virus]. AB - Several methods were employed to obtain lymphocyte cultures from blood samples taken from normal cattle and from cattle affected with enzootic leukosis. Biologic and virologic experiments revealed that the cultures from diseased cattle contained an oncogenic leukosis virus, while those obtained from healthy animals were exempt from such virus. Electron microscopy was applied to study the morphologic aspects of the bovine leukosis virus with a total of 18 lymphocyte cultures. The viral particles noted were shown to have typical configuration and size of type C oncogenic viruses as described in the literature--possessing a central dense nucleotide and double membranes. Seen were also various stages in the development of the virus. Mature virus particles were likewise observed in a stable cell line FLK (percent of the virus) as well as in ultrathin cross sections of the spleen of leukosis-affected animals. PMID- 3024389 TI - [Creation of an avirulent and immunogenic mutant from the rhinopneumonitis virus]. AB - An avirulent immunogenic virus strain mutant of the causative agent of rhinopneumonia was found to cause abortions and respiratory diseases in horses. The mutant was obtained with the use of a virulent strain that induced strongly manifested clinical symptoms of the disease, and was cultured in cell media containing 5-iodine-2-desoxiuridine as an antimetabolite, following a definite pattern. It was found that the mutant completely lost its virulence, however, it retained its immunogenicity. It likewise retained these newly acquired biologic properties with regard to its being stable and irreversible. Animals vaccinated with this mutant did not act as carriers, neither did they shed any virus. The mutant is now used in the production of a vaccine for the practice. PMID- 3024390 TI - Transferrin receptor (TrfR) expression in breast carcinoma and its possible relationship to prognosis. An immunohistochemical study. AB - TrfR, a primitive membrane protein was demonstrated by immunohistochemistry in 87.6% of 105 cases of breast carcinoma, predominantly on the cell surface and in a strong and rather uniform pattern. Sporadic staining in a patchy fashion was observed. No difference between individual tumour types was seen, neither in cytomorphological staining pattern nor in staining intensity. Exceptionally, mucoid carcinomas showed weaker intensity for receptor expression. Because of the heterogenous expression of TrfR within most of the tumours the extent of staining reaction was determined by semiquantitative grading (low, moderate, high). These results were compared with grade of anaplasia, tumour staging and nodal status of the axilla. The extent of immunoreactivity revealed significant correlation with grade of anaplasia, whereas no correlation was found with staging and status of axillary lymph nodes. Tumours with higher degree of malignancy (GII-GIII) showed a higher extent of staining. The presence of TrfR in a high degree of expression thus implies some prognostic value. Its quantitative determination can provide kinetic data on the neoplasm. PMID- 3024391 TI - The nucleotide sequence of the gB glycoprotein gene of HSV-2 and comparison with the corresponding gene of HSV-1. AB - The nucleotide sequence of the gB glycoprotein gene of HSV-2 has been determined and compared with the homologous gene of HSV-1. The two genes are specified by the same total number of codons (904); eight additional codons of the HSV-1 gene are found within the signal sequence, and eight additional codons of the HSV-2 gene are found at three different sites in the gene. The signal cleavage, membrane-spanning, and eight potential N-linked oligosaccharide sites, as well as 5'- and 3'-regulatory signals are largely conserved. The overall amino acid homology is 85%; least conserved are the N- and C-terminal regions of the protein. Secondary structure plots were determined for the two proteins, and the structures were compared with each other and with alterations in structure due to several mutations in the HSV-1 gB gene for which sequence analysis is available. The high homology in primary and secondary structure suggests a conserved, essential function for the gene. PMID- 3024392 TI - Characterization of the chimeric SV40 large T antigen which has a membrane attachment sequence of polyoma virus middle T antigen. AB - A chimeric SV40 mutant, pMTPY, was constructed which codes for a large T antigen having the putative membrane attachment sites of polyoma virus middle T antigen at the carboxy-terminal portion. The mutant T antigen was detected exclusively in the cytoplasm of CV-1 cells transfected with pMTPY by a fluorescent antibody test. This mutant could not support viral DNA replication, but could immortalize secondary cultured rat brain (RB) cells. Immortalized RB cells produced nonkaryophilic large T antigen and also small T antigen. The amount of p53 expressed in those cells was larger than that in control RB cells. In addition, this mutant had the ability to transform NIH3T3 cells. The mutant nonkaryophilic large T antigen in NIH3T3 transformant was localized in cytoplasmic membrane fractions. PMID- 3024393 TI - Expression of gangliosides as receptors at the cell surface controls infection of NCTC 2071 cells by Sendai virus. AB - The involvement of gangliosides as receptors for Sendai virus was established previously using experimentally produced receptor-deficient cells. In the search for a naturally occurring counterpart, NCTC 2071 cells emerged as a likely candidate. These cells in their native state were not agglutinated nor infected by Sendai virus, but were infected by the virus when the gangliosides GD1a, GT1b, or GQ1b were supplied in the culturing medium. Preliminary analysis indicated that NCTC 2071 cells contained an unusually high ratio of sialoglycoproteins to gangliosides. A brief treatment of the cell surface with the protease trypsin made greater than 99% of the native monolayer susceptible to infection by the wild-type virus which contains the viral attachment protein HN. (Incubation of the trypsin-treated cells with a temperature-sensitive mutant missing HN produced no detectable infection.) The increased binding of cholera toxin, a ganglioside specific probe, after incubation of the cells with trypsin and sialidase, was consistent with the hypothesis that gangliosides more complex than GM1 are on the surface of NCTC 2071 cells and that trypsin treatment increases their accessibility. The presence of receptor gangliosides in lipid extracts of NCTC 2071 cells was confirmed by thin-layer chromatography of the ganglioside fraction and by the binding of cholera toxin. These results demonstrate that cells containing receptor gangliosides may still be resistant to infection because these are not expressed properly at the cell surface as receptors for interaction with the HN protein of Sendai virus. PMID- 3024394 TI - Nucleotide sequence and deduced amino acid sequence of the structural proteins of dengue type 2 virus, Jamaica genotype. AB - The nucleotide sequence of the 5'-terminal 2469 bases of dengue 2 (Jamaica genotype) virus has been determined and the encoded proteins compared with those of yellow fever and West Nile viruses, which belong to different flavivirus serogroups. The cDNA clone which was sequenced contains a 5'-noncoding region of 96 nucleotides followed by a single open reading frame coding for the structural proteins 5'-C-prM(M)-E-3' and the beginning of the NS1 nonstructural protein. The amino acid sequence homology between the structural polyprotein precursor of dengue 2 virus and those of yellow fever and West Nile viruses is 36.5 and 42%, respectively. The dengue virus structural proteins are similar in size and composition to those of the other flaviviruses. The basic capsid protein and the membrane and envelope proteins have hydrophobic regions at their C termini. The dengue 2 capsid C, membrane M, and envelope E proteins share 13, 36, and 43% homology, respectively, with the cognate proteins of yellow fever virus, and 33, 32, and 47% homology with the cognate proteins of West Nile virus. All 6 cysteine residues in the dengue 2 premembrane protein and all 12 cysteine residues in the dengue 2 envelope protein are conserved in the cognate proteins of yellow fever and West Nile viruses. PMID- 3024395 TI - Sindbis virus infection increases hexose transport in quiescent cells. AB - Sindbis virus infection of baby hamster kidney cells or chick embryo cells resulted in a significant increase in the rate of uptake of [2-3H]deoxy-D-glucose ([3H]dGlu). Stimulation of hexose transport in Sindbis virus-infected cells occurred only if the cells were rendered quiescent by culturing at high density or by serum starvation. In contrast, Sindbis virus-induced inhibition of potassium transport, measured as a decrease in the uptake of 86Rb+, was independent of cell growth state. Stimulation of [3H]dGlu uptake in Sindbis virus infected cells was the result of an increase in the Vmax of the hexose transporter, but not a change in the Km. The stimulation of [3H]dGlu uptake induced by Sindbis virus was insensitive to the drug actinomycin D, but was blocked by cordycepin. The stimulation was also insensitive to treatment with tunicamycin, which prevented the virally induced inhibition of the plasma membrane-associated Na+/K+ ATPase and termination of host protein synthesis. PMID- 3024396 TI - Generation of cDNA clones of the bacteriophage phi 6 segmented dsRNA genome: characterization and expression of L segment clones. AB - Bacteriophage phi 6 has three dsRNA genome segments of about 3.0, 4.0, and 6.4 kbp. More than 90% of the segmented phi 6 dsRNA genome has been cloned as subchromosomal cDNA fragments, generated by reverse transcription of denatured polyadenylated dsRNA, RNA removal, annealing, filling, size fractionation, tailing, and insertion at the PstI site of pBR322. All of the large (L) segment is represented by five overlapping fragments, 98% of the small (S) segment is present in three fragments, and 67% of the medium (M) segment is contained in two fragments. Fragments have been aligned in linear arrays by Southern blot hybridization and restriction enzyme analysis. The orientation of the ordered fragments with respect to genomic RNA and phi 6 transcriptional direction was determined by comparison of terminal DNA sequences with RNA sequences at the genomic ends of phi 6 RNA. Expression of L segment clones using both Escherichia coli minicells and T7 polymerase/promoter vectors indicate that the order of known phi 6 genes on the large chromosome is: 5'--gene 7, gene 2, gene 4, gene 1- 3'. cDNA complementation of a ts mutant, ts411, has located this mutation in gene 4. PMID- 3024397 TI - Poliovirus temperature-sensitive mutant containing a single nucleotide deletion in the 5'-noncoding region of the viral RNA. AB - The effect on viral replication of deleting nucleotide 10 of the poliovirus RNA genome was determined. This deletion, which removes a base pair from a predicted hairpin structure in the viral RNA, was introduced into full-length cDNA. Virus recovered after transfection of HeLa cells with the mutated cDNA contained the expected deletion and was temperature sensitive for plaque formation. Analysis of viral replication by one-step growth experiments indicated that mutant virus production at the nonpermissive temperature was at least 100 times less than that of wild type virus, and release of virus from mutant-infected cells was delayed. The synthesis of positive- and negative-strand viral RNA in mutant virus-infected cells was temperature sensitive. Virus-specific protein synthesis in mutant virus infected cells was not temperature sensitive but occurred at a slower rate than that of wild type virus at permissive and nonpermissive temperatures. Replication of the mutant virus was sensitive to actinomycin D, in contrast to the wild type parent virus, which was resistant to the drug. Mutant virus stocks contained a small percentage of ts+ viruses that were able to form plaques at the nonpermissive temperature. Nucleotide sequence analysis of genomic RNA from these ts+ viruses revealed a single base change at position 34 from a G to U. In the positive RNA strand, the effect of this mutation is to restore to the hairpin structure the single base pair whose formation was prevented by the original deletion. The ts+ pseudorevertants replicated to similar titers as wild type virus at 33 and 38.5 degrees and were partially sensitive to actinomycin D. PMID- 3024398 TI - Increased sensitivity of virus-infected cells to inhibitors of protein synthesis does not correlate with changes in plasma membrane permeability. AB - Semliki Forest virus-infected BHK cells or herpes simplex virus-infected Vero cells were incubated with the protein synthesis inhibitors hygromycin B and gougerotin. Infected cells take up no more [3H]hygromycin or [3H]gougerotin than do mock-infected cells, at a time p.i. at which either compound is more inhibitory to protein synthesis in infected, than in mock-infected cells. The concentrations of hygromycin and gougerotin required to inhibit protein synthesis in intact cells (irrespective of whether they are infected or not) are several orders of magnitude higher than those required in either permeabilized cells or in cell-free systems. Infected cells take up 86Rb+ at the same rate as mock infected cells, their intracellular content of K+ is the same, and the activity of the Na+ pump is the same. It is concluded that the increased efficacy of hygromycin and gougerotin in virus-infected cells is a consequence of altered intracellular compartmentation and that increases in permeability of the plasma membrane, if any, are so small as to be undetectable by direct methods. PMID- 3024399 TI - The noncoding region of HPV-6vc contains two distinct transcriptional enhancing elements. AB - HPV-6vc subgenomic fragments were inserted into an enhancer-dependent expression vector for chloramphenicol acetyltransferase (CAT) and assayed for the presence of transcriptional enhancing elements. A transcriptional enhancing element was detected in the noncoding region (NCR) of the HPV-6vc viral genome when the CAT assays were performed in viral transformed human kidney cell lines (293 and 324K), in human cervical carcinoma cell lines (HeLa and Siha), and in bovine papillomavirus type 1 (BPV-1) transformed mouse cells (C127-53). The NCR region of the HPV-6b genome was only capable of enhancing transcription of the CAT gene in the HeLa cell line at a level one-third that of the HPV-6vc NCR. The HPV-6vc NCR enhancing activity in C127-53 cells was further stimulated by the addition of sodium butyrate to the growth medium. Localization of the DNA sequences in the HPV-6vc NCR responsible for enhancing transcription revealed two distinct enhancer elements. One element (HPV-6vc position 7218-7544) was active in the 293, HeLa, Siha, and C127-53 cells. The second enhancer element (HPV-6vc position 7544-7971) was only capable of stimulating transcription in HeLa, C127-53, and Siha cells. When the HPV NCR-CAT expression vectors were cotransfected with a competitor plasmid (pNCR75) into C127-53 or HeLa cells then transcriptional enhancement decreased, indicating competition of cellular factors which affect both segments of the HPV-6vc NCR. PMID- 3024400 TI - Selective integration of avian leukosis virus in different hematopoietic tissues. AB - Hematopoietic tissues obtained from avian leukosis virus (ALV)-infected Hyline SC chickens were analyzed for the presence of integrated viral DNA sequences. Cells were prepared from bone marrow, bursa, spleen, thymus, and peripheral blood. Following the removal of erythrocytes, cellular DNAs from each of these tissues were examined by Southern analysis. During the first few weeks of infection, DNA from the bone marrow contained as many as 0.5 copies of viral DNA per haploid genome. Cells from the bursa and peripheral blood contained between 0.05 and 0.15 copies per haploid genome. In contrast, neither splenic nor thymic DNA contained significant levels of viral DNA sequences even though infected birds developed titers of circulating virus between 10(5) and 10(6) IU/ml of plasma. DNA prepared from erythrocytes isolated from the peripheral blood of these birds contained approximately 0.4 copies of integrated viral sequences per haploid genome at 2 weeks after infection. Despite greater levels of integrated sequences in other tissues, by 9 weeks after infection viral sequences were detected only in DNA from bursal lymphocytes. Cells prepared from the spleen and thymus of infected birds were also examined for their size distribution, their internal complexity and their surface expression of immunoglobulin. None of the populations examined differed from normal, uninfected control preparations. These results suggest that ALV infection occurs primarily in the bone marrow and that the different tissues of the hematopoietic system are selectively infected. Further, these results indicate that ALV infection persists longer in bursal lymphocytes than in other hematopoietic tissues. Previous studies have demonstrated that the lymphoid tumors that develop in white leghorn chickens following ALV infection are bursal dependent B-cell lymphomas that express immunoglobulin M. The observations presented in this communication offer, in part, an explanation for the restricted phenotype of the lymphoid tumor that develops in the SC chicken. Further, the data suggest an explanation for the bursal-dependent nature of the ALV-induced lymphoma. PMID- 3024401 TI - Biosynthesis and chemical and immunological characterization of avian reticuloendotheliosis virus env gene-encoded proteins. AB - Two glycosylated proteins designated gp90 and gp20 were purified from replication competent avian reticuloendotheliosis associated virus (REV-A). The N-terminal sequences of gp90 and gp20 were determined and found to match the REV-A-env-gene sequence. The alignments of the determined amino acid sequences with the predicted sequence indicate that gp20 and gp90 are the REV-A-encoded viral transmembrane and surface glycoprotein, respectively, and predict a signal peptide of 36 residues on the 5' end of the env-gene. Furthermore, gp90 of REV-A was detected by Western blot analysis with antibodies to a tridecapeptide corresponding to an env-gene nucleotide segment immediately preceding gp20 and thus representing the C-terminal portion of gp90. The env-gene precursor polyprotein gPr75-79env and Pr22(E), the precursor to gp20 and p2(E) were identified in the infected cells by monospecific antibodies raised against purified gp20. Thus the organization of gPR75-79env is likely to be N-gp90-gp20 p2(E), resembling that of M-MuLV gp85env. Sequence comparisons showed that the env gene of REV-A is highly related to both baboon endogenous virus and Type D retroviruses. In Western blot analyses, antibodies to REV-A gp20 cross-reacted with a panel of mammalian Type C and Type D viruses. Evolutionary aspects of these findings are discussed. PMID- 3024402 TI - Butyrate-induced reversal of herpes simplex virus restriction in neuroblastoma cells. AB - The synthesis of herpes simplex virus (HSV) in mouse neuroblastoma cells (NB, clone 41A3) is restricted. There was a disappearance of infectious virus upon serial passage of infected cells. NB cells treated with sodium-n-butyrate for 24 hr before infection synthesized 200-2000 times more HSV than untreated cells. Infectious center assays demonstrated that the number of cells capable of producing HSV was increased as a result of butyrate pretreatment. Although host protein synthesis was inhibited by HSV infection, viral-induced protein and DNA syntheses were not detected in the absence of butyrate. Cycloheximide blocked the induction of permissiveness by butyrate suggesting that a protein(s) was responsible for allowing HSV synthesis in NB cells. Regulatable host factors involved in HSV replication in neural cells can be studied in the system described. PMID- 3024403 TI - Herpesvirus (pseudorabies virus) latency in swine: occurrence and physical state of viral DNA in neural tissues. AB - The occurrence of the pseudorabies virus (PRV, herpes suis 1) genome in various neural tissues of latently infected pigs was investigated. During the latent phase of infection, between 7 and 52 weeks p.i., the average amount of PRV DNA ranged between 0.3 and 0.05 genome copies per cell. The results obtained by in situ cytohybridization and reassociation kinetic experiments indicated that each latently infected cell harbored at least 30 viral genome copies. PRV DNA could be demonstrated in similar frequencies (about 30% of cases) in the trigeminal ganglia, the olfactory bulb, and the medulla oblongata, and less frequently in the brain stem and the spinal cord. Southern blot analysis showed that in general the physical state of the latent genome was linear and nonintegrated. Only in 2 of 15 animals could the presence of circular or concatemeric viral DNA be observed. Thus, we could show that over a period of 13 months after infection the PRV genome persisted both qualitatively and quantitatively in a stable state in different areas of both the peripheral and the central nervous system. PMID- 3024404 TI - Coding strategy of the S genome segment of Hantaan virus. AB - Hantaan virus is the type species of the recently recognized Hantavirus genus of Bunyaviridae. The small (S) RNA segment of the negative-sense, tripartite genome was molecularly cloned and the nucleotide sequence was determined. The RNA sequence derived from the cDNA copy was found to contain 1696 nucleotides. A single open reading frame of sufficient size to encode the virus nucleocapsid protein was detected in the cDNA corresponding to viral complementary-sense RNA. RNA transcripts of the cDNA were synthesized with SP6 polymerase and were used to program cell-free reticulocyte lysate translation systems. Viral complementary sense transcripts served as efficient messages in translation systems and generated Hantaan nucleocapsid protein. No translation products were detected when lysates were programmed with viral-sense transcripts. This coding assignment of the nucleocapsid protein to the viral complementary-sense RNA of the S genome segment is consistent with those of other members of this family. Unlike other Bunyaviridae, which encode both a nucleocapsid protein and a nonstructural (NSs) protein of similar sizes, a NSs protein has not been identified for Hantaan virus. Furthermore, other than the nucleocapsid protein gene sequence, the only potential open reading frame in Hantaan S RNA encoded a short, 48-amino acid polypeptide which initiated two codons beyond the termination of the nucleocapsid protein in the same reading frame. These data demonstrate that the coding strategy of the Hantaan virus S RNA is different than those reported for other viruses in this family. PMID- 3024405 TI - Characterization of subviral particles in cells infected with simian rotavirus SA11. AB - Subviral particles were isolated from lysates of simian rotavirus SA11-infected cells by sedimentation through sucrose gradients and separated by equilibrium centrifugation in CsCl gradients. A cell-free system that supports rotavirus RNA replication and transcription was used to identify particles in the CsCl gradients with associated polymerase activity. These data indicated that particles with densities of 1.34 and 1.38 g/cm3 were responsible for most of the transcriptase activity present in infected cells. Electrophoretic analysis showed that particles at 1.34 g/cm3 were analogous to double-shelled virus, consisting of the inner shell proteins VP1, VP2, and VP6, the outer shell proteins VP3 and VP7, and DS RNA. Particles of 1.38 g/cm3 were similar to single-shelled virus containing the inner shell proteins and DS RNA. The pellets of the CsCl gradients were enriched for subviral particles with replicase activity. Analysis of the pellets suggested that replicase particles contain a core of VP1 and VP2 that is similar to that found in single- and double-shelled virus but contain significantly less VP6 protein per particle than those with transcriptase activity. Two particles were detected in infected cells that contain no detectable polymerase activity; one consisted primarily of the structural proteins VP2, VP3, and VP6 and the other of the nonstructural protein NS35. PMID- 3024406 TI - Monoclonal antibodies to the v-fes product and to feline leukemia: virus P27 interspecies-specific determinants encoded by feline sarcoma viruses. AB - Monoclonal antibodies to p27 gag and v-fes specific determinants on the gag-onc poly-protein encoded by Snyder-Theilen feline sarcoma virus (ST-FeSV) were prepared. In order to obtain hybridoma clones specific to the antigenic determinants encoded by the FeSV genome, Lou rats were immunized with ST-FeSV transformed, virus-nonproducing syngeneic cells, and boosted with either the same cells or affinity-purified feline leukemia virus (FeLV) p27. Three distinct clones reactive to both FeLV p27 and p85gag-fes, and one clone specific for a p85fes determinant were established. The anti-p27 monoclonal antibodies also reacted with the polyproteins p95gag-fes and p83gag-fgr, from Gardner-Arnstein (GA) and Theilen-Pedersen (TP1) FeSV, respectively. The anti-p27 monoclonal antibodies reacted with MuLV p30 and RD114 p28 but not with RSV, MMTV, or BLV. These results indicated that the part of the p27 gag gene that is preserved in ST , GA, and TP1-FeSV encodes interspecies-specific p27 determinants. PMID- 3024407 TI - Differences in bovine parainfluenza 3 virus variants studied by monoclonal antibodies against viral glycoproteins. AB - We previously showed that of three bovine parainfluenza 3 virus strains the M strain, which is neurovirulent for young mice, has an extensive syncytium inducing activity, whereas avirulent SC and 910N strains are weak in this activity. It was also demonstrated that both M and SC strains have very low hemagglutination and neuraminidase activities, while the 910N strain shows these activities to high levels. In the present study, monoclonal antibodies (Mabs) were raised against the glycoproteins of the 910N strain, and utilized to further characterize these three viral strains. Five Mabs against the hemagglutinin neuraminidase protein, which were classified into four different epitope recognizing groups, neutralized the M strain much more effectively than the 910N and SC strains, while the Mabs showed lower hemagglutination inhibition (HI) titers against the M and SC strains than the 910N strain. Three Mabs against the fusion protein neutralized the M strain but not the 910N and SC strains, while they showed no HI activity against any of these strains. These findings suggested that the M strain is considerably different from other strains in the structure of the viral envelope proteins. PMID- 3024408 TI - The processing of pseudorabies virus glycoprotein gX in infected cells and in an uninfected cell line. AB - Pseudorabies virus (PRV) produces a glycoprotein, gX, that accumulates in the medium of infected cells. The gX gene was expressed in Chinese hamster ovary cells (CHOgX cells) using the cytomegalovirus Towne major immediate early promoter. Like PRV-infected cells, CHOgX cells produced gX and exported it into the medium. Tunicamycin reduced the molecular weight of the gX in the medium to 89 kDa, compared with 99 kDa for gX made in the absence of drug. In the presence of tunicamycin gX produced by both PRV-infected cells and CHOgX cells was still glycosylated, as indicated by incorporation of [14C]glucosamine. The most likely form of this glycosylation is O-linked. In a pulse-chase experiment, gX first appeared in a 90-kDa form, then a 115-kDa form. This 115-kDa form is probably cleaved to give the 99-kDa form of gX that is released into the medium. The 115 kDa form was much more persistent in the PRV-infected Vero cells than in the CHOgX cells. In both cell types, gX was labeled by [35S]sulfate in the presence and absence of tunicamycin. PMID- 3024409 TI - Infection of the HTLV-II-bearing T-cell line C3 with HTLV-III/LAV is highly permissive and lytic. AB - The recent discovery that HTLV-I infected T-cells have an enhanced susceptibility to HTLV-III/LAV infection with concurrent cell lysis (S. Harada, Y. Koyanagi, and N. Yamamoto (1985) Science, 229, 563-566) led us to investigate a possible role for HTLV-II infected T cells in this respect. HTLV-II-bearing C3 cells were exposed to HTLV-III/LAV and the subsequent infection monitored by indirect immunofluorescence (IIF) of cells, reverse transcriptase (RT) activity in culture fluids, and hemacytometer cell counts. HTLV-III/LAV antigen expression was detected within 2 days whereas RT activity was detected within 3 days postinfection. The presence of immunofluorescent positive cells and RT activity was accompanied by cell lysis. Double infection of C3 cells was confirmed by IIF using a goat polyclonal antibody directed against the core p24 protein for detecting HTLV-II, and high titer serum from a pre-AIDS patient for detecting HTLV-III/LAV. We conclude that HTLV-I and HTLV-II infected T-cells share an enhanced susceptibility to HTLV-III/LAV infection with the promotion of cell lysis. PMID- 3024410 TI - Characterization of hepatitis A virus structural proteins. AB - HAV particles isolated from infected cells banded at buoyant densities of 1.42, 1.32, and 1.20 g/ml, and distinctive protein patterns were established by gel electrophoresis and reverse phase high performance liquid chromatography. The relatively higher amounts of p30 in particles with lower buoyant densities suggest that this protein is VP0 and is part of the immature picornavirion. The protein elution profiles obtained by HPLC were virtually identical for all the HAV strains examined but differed from those of other picornaviruses. The N terminal amino acid sequence of VP1 and VP2 was determined and aligned to the nucleotide sequence. Sequencing VP0 and VP3 was not possible, probably because the amino termini are blocked. VP1, VP3, and VP0 induced specific antibodies in rabbits. PMID- 3024411 TI - Expression of endogenous retroviral envelope glycoprotein as a determinant of immunity to Rous sarcoma. AB - To analyze the effect of the expression of endogenous retroviral envelope glycoprotein on tumor immunity, patterns of sarcoma growth were compared in inbred FP line chickens infected with either of two strains of avian sarcoma virus, Pr-B (subgroup B) or cl.85 (subgroup G). These viruses were chosen for analysis because the envelope glycoprotein of Pr-B, but not of cl.85, is antigenically cross-reactive with the endogenous retroviral envelope glycoprotein expressed in the FP line. Inoculation of 1-day-old hatchmates with either virus yielded a significant percentage of chickens with distal sarcomas localized to visceral organs. By contrast, a marked difference in the percentage of chickens bearing distal sarcomas was noted when sarcoma tissue excised from virus inoculated donors was implanted in 1-day-old recipients; a high proportion of the recipients of Pr-B-induced sarcoma tissue (Pr-B-sarcoma recipients), but only a low proportion of the cl.85-sarcoma recipients, exhibited distal sarcomas. At 3 weeks posthatch, a significantly higher percentage of donor-derived cells was detected in the primary tumors of the cl.85- versus the Pr-B-sarcoma recipients. A model of immune control, premised on the tolerogenicity of endogenous viral glycoprotein, is proposed to rationalize these results. PMID- 3024412 TI - Isolation and characterization of covalently closed circular proviral DNA molecules of several type D retroviruses isolated from human cell lines. AB - Infection of a human lymphoblastoid B cell line (Raji cells) with type D retroviruses, originally isolated either from subhuman primates (MPMV, LV) or from permanent human cell lines (PMFV, HeLaV, HEp-2V) led to the production of type D retrovirus particles. Subsequent cocultivation of uninfected and virus producing Raji cells was employed for the generation of sufficient amounts of covalently closed circular DNA molecules (cccDNA). Highest amounts of cccDNA were obtained after cocultivation of virus-producing Raji cells and homologous uninfected cells at a ratio of 1 to 3 for 72 hr. The cccDNAs of type D retroviruses migrated at about 4.3 kbp compared to lambda DNA/HindIII markers. Digestion of cccDNAs with restriction endonucleases which have one recognition site generated molecules of approximately 8 kbp. The restriction endonuclease site analysis of the cccDNA of type D retroviruses revealed a genomic heterogeneity among the different isolates. PMID- 3024413 TI - [Effect of chemotherapy on the nucleoside phosphate kinase activity in experimental tumors]. AB - In animals with experimental transplantable tumors, an effective treatment with antitumor compounds brought about interrelated changes in nucleoside phosphate kinase activity. The decrease in thymidine phosphate kinase activity was as a rule matched by an increase in that of uridine phosphate kinase. Consequently, "thymidine kinase-uridine kinase" shunt responsible for thymidine-uridine interconversion was suggested, which is switched on when the homeostasis of tumor cells is disturbed. In treatment of tumor-bearing animals, the effective dosage of antitumor drugs was considerably decreased due to a combination of the said compounds (inhibiting nucleoside kinases) or with azauridine which inhibits uridine monophosphate synthesis from orotidine-5'-phosphate. PMID- 3024414 TI - [Hygienic standards for the concentration of quartz and coal dusts in the air of a work area (a review of the literature)]. PMID- 3024415 TI - Applying information technology to clinical medicine. PMID- 3024417 TI - [How dangerous is NMR tomography?]. PMID- 3024416 TI - [Immunohistochemical studies in the differential diagnosis of malignant fibrous histiocytoma]. AB - Malignant fibrous histiocytomas (MFH) belong to the most frequent soft tissue tumours in adults and have to be discriminated from other tumours with similar morphology. Various tumour markers aid the differential diagnosis. Twenty cases of MFH were studied immunohistochemically using antibodies to vimentin, TPA, desmin, lysozyme, alpha 1-antitrypsin, alpha 1-antichymotrypsin, S-100 protein, neurone-specific enolase (NSE), laminin, fibronectin and ferritin. Vimentin and lysozyme were found in the tumour cells of all, alpha 1-antitrypsin of 18, alpha 1-antichymotrypsin of 19, fibronectin of 16 and ferritin of 12 cases. Antibodies of TPA, desmin, S-100 protein, NSE and laminin did not reveal positive immunoreactivity. Exclusion of spindle-cell carcinoma can be made by positive vimentin and negative TPA reactivity, of melanoma by negative S-100 reactivity, and of leio- and rhabdomyosarcoma by lack of desmin immunoreactivity. Schwannomas contain S-100 protein, but lack lysozyme, alpha 1-antitrypsin, alpha 1 antichymotrypsin and fibronectin. Pleomorphic liposarcomas cannot be distinguished from MFH on the basis of immunohistochemical staining. Vimentin, alpha 1-antitrypsin, alpha 1-antichymotrypsin and fibronectin can, therefore, be regarded as useful markers in the differential diagnosis of MFH. PMID- 3024418 TI - [Pathogenetic mechanisms in autoimmune diseases]. AB - In addition to an earlier review [Z. ges. inn. Med. 35 (1980) 58] newer concepts in the pathogenesis of autoimmune diseases are presented. Acantholysis in pemphigus is caused by proteases released from cells of stratified squamous epithelium stimulated by antibodies to cell membrane antigens. The dermoepidermal cleavage in pemphigoid is presumably due to proteases from granulocytes accumulating at the dermoepidermal junction by antibody-mediated complement activation. Receptor autoantibodies can directly stimulate or block receptor function, increase cell membrane receptor degradation, or activate complement with lysis of receptor-carrying membrane structures. Tissue injury in immune complex diseases is caused by reactive oxygen metabolites and proteases released from stimulated phagocytes. PMID- 3024419 TI - Towards a selective chemotherapy of virus infections. Development of bromovinyldeoxyuridine as a highly potent and selective antiherpetic drug. PMID- 3024420 TI - [Function of the hypophyseo-adrenal system in patients with psoriasis and psoriatic arthritis]. PMID- 3024421 TI - [The sinusoid wall of the liver in viral infections]. AB - The elements of the "peri-sinusoidal functional complex" (i.e. non-hepatocytes, space of Disse, vascular pole of hepatocytes) can react to the presence of virus in the sinusoidal blood stream by retaining viruses, sometimes followed by viral proliferation, by reactive proliferation, or by degenerative changes in the cells which may culminate in cell death. Immune processes may be also activated or alternatively, there may be no response whatever to the viremia. In hepatitis B infection the principal changes are in the Kupffer cells and in the vascular pole of the hepatocytes. Along with immunologic responses there are proliferative and degenerative events in the Kupffer cells, which may deleteriously affect the sinusoidal cell membrane of the hepatocytes. These factors, as well as the receptor structure of these cell membranes, are regarded as responsible for the uptake of virus in the hepatocyte. Cytotoxic T-cells which have migrated to the region also play a role in the overall process. The point should be made, however, that these concepts are largely hypothetical and that hepatitis B virus particles have yet to be demonstrated in the peri-sinusoidal complex. PMID- 3024422 TI - [Experiences with determining carcinoembryonic antigen in tumors of the digestive tract]. AB - CEA levels (CEA = carcino-embryonic antigen) were checked as indicators for the presence of malignant diseases of the gastro-intestinal tract. Tests were applied to 1,102 blood samples, including 1,012 relating to malignant alterations of the digestive tract and 32 relating to benign changes. Sixty-eight samples were taken from tumours localised elsewhere. The importance is stressed of high-continuity observation of CEA levels which have proved to enable dimensional assessment of colorectal carcinoma. Presurgical and postsurgical CEA values can be related to surgical radicality. Regular postoperative monitoring is recommended for prognostication, early detection of recurrences, and early assessment of spreading of the disease. PMID- 3024423 TI - [The surgical abdomen in AIDS patients]. PMID- 3024424 TI - Modulation of Bhanja virus infection in mice. AB - Randombred (ICR) and inbred (C57B1/6) 4-wk-old SPF male mice were infected extraneurally with Bhanja virus (Bunyaviridae) and subjected to various treatments. Immunosuppression with cyclophosphamide (CPA) affected the course of the infection when a higher dose (10(6) suckling mouse intracerebral LD50) of the virus and 2 or 3 injections of CPA (150 mg/kg each) were given: then a part of the animals died due to viral encephalitis, whereas all the CPA-untreated infected mice survived. A dual peripheral infection with Bhanja and Lipovnik (Reoviridae) viruses did not cause any symptomatic response, and the host's humoral antibody was slightly stimulated. When Bhanja virus was given prior to, or simultaneously with, tick-borne encephalitis virus (Flaviviridae), a moderate decrease of the mortality (due to tick-borne encephalitis) occurred. A mixed peripheral infection of mouse with Bhanja virus and Cryptococcus neoformans, did not result in a fatal virus encephalitis of the host, nor was cryptococcosis affected substantially. However, formalin-killed cells of the fungus ("cryptococcin") administered before the extraneural inoculation of Bhanja virus caused an 8-fold increase of antibodies neutralizing the virus; a mild therapeutic or protective effect of cryptococcin on encephalitis after an intracerebral application of Bhanja virus was also observed. PMID- 3024425 TI - [Further studies with the 3:3 procedure in the diagnosis of leukosis and thoughts on a change in leukosis regulations]. PMID- 3024426 TI - Effect of a heme-peptide derived from cytochrome-c on lipid peroxidation. II. Experiments with liver microsomes. AB - ADP-Fe2+ stimulated, NADPH dependent lipid peroxidation of liver microsomes (as measured by malondialdehyde formation) was not inhibited by c-heme-nonapeptide, unlike the same process in brain microsomes. However, in the presence of 5 mM aminopyrine (causing partial inhibition) or SKF-525A (a specific inhibitor of cytochrome P450) the residual activity of lipid peroxidation of liver microsomes was markedly inhibited by c-heme-nonapeptide. Further, c-heme-nonapeptide itself prevented the transient accumulation of lipid hydroperoxides during ADP-Fe2+ stimulated lipid peroxidation. These results led us to suggest two different pathways of lipid peroxidation. The first route involves cytochrome P450. The second pathway, which can be inhibited by c-heme-nonapeptide, appears to be more important physiologically. PMID- 3024427 TI - Effect of a heme-peptide derived from cytochrome-c on lipid peroxidation. I. Effects on brain microsomes. AB - Heme-nonapeptide inhibits NADH and NADPH dependent lipid peroxidation of brain microsomes in the presence or absence of ADP-Fe complex. The transient accumulation of lipid peroxides during NADH or NADPH dependent, ADP-Fe stimulated lipid peroxidation, is inhibited by heme-nonapeptide. Oxygen consumption of brain microsomes in the presence of NADH or NADPH is stimulated by heme-nonapeptide. Reduction of cytochrome-c and nitro-tetrazolium-blue by O2- generated by xanthine oxidase is inhibited by heme-nonapeptide. PMID- 3024428 TI - [An uncommon course of Paget's disease]. PMID- 3024429 TI - Use of biotinylated DNA probes in screening cells obtained from cervical swabs for human papillomavirus DNA sequences. AB - A nonradioactive DNA-detection procedure using biotinylated DNA probes in the screening of cells from cervical swabs for DNA sequences homologous to human papillomavirus (HPV) DNA was tested. This alternative DNA-detection method yielded results comparable to those obtained with radioisotope-labeled DNA probes in 32 cases tested. This procedure obviates the special precautions required for radioisotope materials. Accordingly, this technique can be made available to many laboratories, and conclusive evidence as to the relation of HPV infection to cervical cancer may thus be accumulated. PMID- 3024430 TI - Cerebrospinal fluid cytology diagnosis of meningeal carcinomatosis in patients with small-cell carcinoma of the lung. A study of interobserver and intraobserver variability. AB - The interobserver and intraobserver variation in the cytologic diagnosis of malignancy was determined in 62 cerebrospinal fluid (CSF) specimens from 46 patients with small-cell carcinoma of the lung. In all patients, lumbar puncture was carried out because of suspected central nervous system metastases. Forty CSF specimens from 26 patients with meningeal carcinomatosis and thus with a high probability of a positive CSF cytology were mixed with 22 specimens from 20 patients without meningeal carcinomatosis. The slides were evaluated blindly by two observers, one of whom evaluated all specimens on two separate occasions; only positive, negative and suspicious conclusions were permitted. The consistency of the intraobserver and interobserver conclusions on the initial CSF specimen in each case was 87%. In 13% of the initial CSF specimens in each case, a suspicious conclusion was reached in one of the three evaluations. For all 62 CSFs, the intraobserver and interobserver disagreement was 2% and 3%, respectively. In the first and second evaluations by the one observer and the single evaluation by the other, 17 (65%), 15 (58%) and 12 (46%), respectively, of the 26 "high probability" patients were found to have malignant cells in the CSF. CSF cytology was negative in all 20 patients without meningeal carcinomatosis. Of 10 patients with autopsy-proven meningeal carcinomatosis, 40% were not diagnosed while alive. Multiple CSFs from repeated lumbar punctures increased the number of positive evaluations by 30%. At least 60% of those patients with a suspicious CSF cytology did in fact have meningeal carcinomatosis. On the other hand, 30% of the patients with a positive lumbar puncture had a subsequent negative one. PMID- 3024431 TI - Bronchioloalveolar carcinoma presenting with meningeal carcinomatosis. Cytologic diagnosis in cerebrospinal fluid. AB - The cytologic findings in a 35-year-old patient with bronchioloalveolar carcinoma who initially presented with central nervous system involvement are reported. Following the cytologic diagnosis of carcinomatous meningitis (metastatic adenocarcinoma), an open lung biopsy was performed, which confirmed the presence of a primary pulmonary neoplasm (bronchioloalveolar carcinoma). This case illustrates the importance of the cytologic diagnosis of a clinically unsuspected primary neoplasm. Further, together with three earlier reported cases, it indicates that, in young patients, tumor cells shedding into the cerebrospinal fluid can be the first indication of bronchioloalveolar carcinoma. PMID- 3024432 TI - Malignant pericardial effusion and cardiac tamponade. AB - Cardiac tamponade due to malignant effusion, though rarely the initial manifestation of malignancy, is usually secondary to adenocarcinoma of the lung. Two cases are reported. One patient presented with cardiac tamponade; the other had diffuse cutaneous involvement of the left neck and shoulder two months before he presented with cardiac tamponade. Cytologic examination of both fluids revealed adenocarcinoma. Ultrastructural examination showed poorly differentiated adenocarcinoma in the first patient and bronchioloalveolar carcinoma in the second; carcinoembryonic antigen levels in the fluids were 9.4 ng/mL and over 60 ng/mL, respectively. The computed tomographic (CT) scans of both patients revealed mediastinal fullness with no lung involvement. Even in the absence of a pulmonary mass, lung carcinoma may be the likely primary in patients with malignant pericardial effusions. PMID- 3024433 TI - Fine needle aspiration cytology of sarcomas metastatic to the lung. AB - A review was made of the morphologic features of cells aspirated from 17 sarcomas (5 malignant fibrous histiocytomas, 3 fibrosarcomas, 3 leiomyosarcomas, 3 endometrial stromal sarcomas, 1 osteosarcoma and 2 poorly differentiated sarcomas) metastatic to the lung, paying particular attention to the morphologic differences between the cells of sarcoma and carcinoma and between the cells of the different types of sarcoma. In general, sarcomas were characterized by loosely cohesive, rather flat, cellular aggregates and isolated cells. Three dimensional cell balls or clusters were not present in any case. Cellular pleomorphism was a common, though not invariable, feature. Each type of sarcoma showed some morphologic distinctiveness; however, certain morphologic features were common to more than one type of sarcoma. By comparing the cytologic features of metastatic sarcomas in aspirates with the histologic features of the primary neoplasms, it should usually be possible to decide if a pulmonary lesion is a metastatic sarcoma. PMID- 3024434 TI - Effect of vitamin D3 loading and thyroid hormone replacement therapy on the decreased serum 25-hydroxyvitamin D level in patients with hypothyroidism. AB - Twelve hypothyroid subjects, 13 healthy and 12 healthy women with a slight deficiency of vitamin D were studied to distinguish seasonal changes from the thyroxine-dependent ones. Serum 25-hydroxyvitamin D levels of hypothyroid patients were lower than those of healthy individuals when the sera were obtained in the autumn. In hypothyroid patients a single oral dose of 100,000 IU vitamin D3 resulted in a smaller increase in 25-hydroxyvitamin D concentration than in controls having subclinical exogenous vitamin D deficiency. Substitution therapy with thyroid hormone, started in our study always in autumn, increased the 25 hydroxyvitamin D concentration in hypothyroid patients, which was opposite to the autumn-to-spring variation of this hormone observed in healthy controls. The increase of 25-hydroxyvitamin D, dehydroepiandrosterone and its sulphate values following substitution therapy in the hypothyroid patients may indicate that thyroid hormone(s) is (are) involved in the regulation of steroid hormone synthesis. PMID- 3024435 TI - Stimulation of thyroid adenylate cyclase activity by sera from patients with non thyroidal illness. AB - We have previously shown that sera from many hypothyroid patients stimulated adenylate cyclase activity as measured by serum bioactive TSH concentrations produced by FRTL-5 cell line. This TSH-stimulating activity did not correlate with serum immunoreactive TSH. IgG fractions of these sera did not stimulate FRTL 5 cells. The present study was, therefore, undertaken to investigate the thyroid stimulating activity of sera from patients with non-thyroidal illness. Studies were performed in 36 patients with various non-thyroidal illness. In these patients, serum concentrations of T4, free thyroxine, T3, and TSH were determined. In addition, sera were incubated with FRTL-5 cells or porcine thyroid cells in primary culture in the presence of 0.4 mM MIBX, and medium cAMP concentrations were determined by radioimmunoassay. Sera obtained from some patients with various non-thyroidal illness increased cAMP concentrations in culture media of FRTL-5 cells as well as that of porcine thyroid cells. The thyroid stimulating effects of sera were not disease specific and significantly correlated inversely with serum T3 and T4 concentrations. Serum TSH concentrations in these patients were within the normal range even by the newly developed ultrasensitive assay. Although the nature of substance(s) present in sera of patients with low T3 syndrome which stimulates thyroid adenylate cyclase is not entirely known, it is conceivable that there exist mechanisms independent of TSH to compensate the decreased serum T3 levels in low T3 syndrome. PMID- 3024436 TI - Transient lack of response to TSH of human cultured thyroid cells obtained from hyperfunctioning tissue. AB - TSH-induced cAMP accumulation in cells obtained from normal and pathological thyroid tissue was studied during the first 12 days of primary culture. In normal thyroid tissue cultures (N = 7), the response of cAMP to TSH was present from the second day of culture and reached its maximum after 8 days. A similar behaviour was observed in cultures obtained from euthyroid sporadic goitres (N = 8), even if the rate of response was slightly lower than that of normal tissue. Similarly, cultured cells from euthyroid 'autonomous' nodules (N = 8) appeared to be responsive to TSH during the period of study, but the rate of response was also lower than in the controls. On the contrary, in cultures obtained from toxic adenomas (N = 5) and from diffuse toxic goitres (N = 5) the response to TSH was absent during the first 4 days of culture. The cells became sensitive to TSH from 6 and 6 day onwards, with the rate of response increasing progressively and reaching its maximum on day 12. Finally, in cultured cells obtained from different areas of multinodular toxic goitres (N = 4), the response to TSH was similar to that of euthyroid goitres in cells prepared from 'cold' areas, and to that of toxic adenomas in cells obtained from 'hot' areas. The present data demonstrate the existence of an inhibitory action of unknown factors, possibly iodothyronines or thyroglobulin, on the TSH effect in short-term cultures obtained from thyrotoxic tissues. A normal TSH responsiveness can be restored when the culture is prolonged. PMID- 3024437 TI - Steroidogenic activity of highly potent melanotropic peptides in the adrenal cortex of the rat. AB - Highly purified ACTH and MSH peptides were studied in isolated rat glomerulosa and inner zone cells and their activity compared with that in an Anolis melanophore assay. While both ACTH1-39 and ACTH1-24 were almost equally potent steroidogenic peptides in the two cell types (ED50 between 1 and 4 X 10(-12) M), alpha-MSH displayed only weak steroidogenic activity. Although it was a full agonist, it was about 10(4)-fold less potent in both capsular and inner zone cells. beta-MSH (porcine) was even 10-fold less active in capsular cells than alpha-MSH, and in inner zone cells it was a partial agonist. Highly potent melanotropic peptides, such as (Nle4, D-Phe7)-alpha-MSH or cyclic (Cys4, Cys10) alpha-MSH were either inactive or exhibited only a very slight partial steroidogenic activity in both cell types. Comparison of the activity profile of additional compounds, such as des-acetyl alpha-MSH, (Tyr(I)2)-alpha-MSH, (Trp(For)9)-alpha-MSH or (Nva12)-alpha-MSH, in the adrenocortical and pigment cell assays led to the conclusion that alpha-MSH does not exert its steroidogenic effect through a typical melanocyte-type of MSH receptor, but rather through a low affinity-type of ACTH receptor. PMID- 3024438 TI - Interaction of PGF2 alpha with prolactin on the release of cyclic AMP and progesterone from the isolated perfused ovary of the pregnant mare serum gonadotropin-treated rat. AB - A newly developed model for perfusion of the isolated rat ovary was employed to study the interactions of Prl with PGF2 alpha in respect to the effects of LH on cAMP formation and progesterone production in the 5 day old corpus luteum of the PMSG-treated rat. An inhibitory effect of PGF2 alpha on both basal and LH stimulated progesterone secretion was found. This block also involved inhibition of the ovarian cAMP release which was not associated with a reduction of the flow of the medium to the ovary. When Prl was present in the medium the PGF2 alpha block of LH-induced cAMP release was reversed. However, Prl failed to restore block of LH stimulation of progesterone secretion in 4 out of 9 experiments, indicating an additional site of action of PGF2 alpha distal to the cAMP in these experiments. PMID- 3024439 TI - In vivo evidence for dopaminergic regulation of the canine pituitary intermediate lobe. AB - In order to examine regulation of pituitary intermediate lobe secretion, plasma immunoreactive (i)ACTH, cortisol, and alpha-MSH responses to iv bolus injections of CRF, quipazine maleate (serotonin agonist), isoproterenol (beta-adrenergic agonist) or haloperidol (dopamine antagonist) were determined in conscious, unrestrained dogs. Endocrine responses to these test substances were also determined in dogs pre-treated with dexamethasone. Administration of one or more doses of each test substance resulted in significant elevations in plasma iACTH and cortisol concentrations. Only haloperidol injection caused significant increases in plasma i alpha-MSH. Following dexamethasone pre-treatment, plasma iACTH and cortisol increases in response to all test substances were considerably reduced or abolished. Dexamethasone did not alter baseline or haloperidol stimulated plasma i alpha-MSH concentrations. However, infusion of bromocriptine mesylate (dopamine agonist) in combination with dexamethasone pre-treatment reduced the plasma i alpha-MSH response to haloperidol. We conclude that a dopaminergic pathway is important in the in vivo regulation of pituitary intermediate lobe activity in dogs. PMID- 3024440 TI - Changes in the responsiveness of perifused rat adenohypophysial cells to repeated stimulation with luteinizing hormone releasing hormone. AB - The ability of luteinizing hormone releasing hormone (LRH) to stimulate the release of luteinizing hormone (LH) from columns of enzymatically dispersed perfused adenohypophysial cells is being used to study the mechanisms controlling the secretion of LH. LRH stimulated the release in vitro of LH from columns of rat pituitary cells. However, when exposed repeatedly (1 pulse every 12 min) to the same submaximal dose (8 nmol/l) of LRH the cells always exhibited a marked progressive increase and subsequent decrease in their responsiveness. Similar effects occurred when the interval between pulses was extended to 20, 30 or 45 min. The enhanced responsiveness of the cells was prevented by the inclusion of protein synthesis inhibitors, cycloheximide or puromycin, in the perifusion fluid. Cells removed from rats ovariectomized 14 days previously also failed to exhibit increased responsiveness when stimulated repeatedly with LRH. LH secretion was also elicited by K+ (50 nmol/l), 8-bromoadenosine 3'-5'-cyclic monophosphate (8-Br-cAMP, 6 nmol/l), 8-bromoguanosine 3'-5'-cyclic monophosphate (8-Br-cGMP, 6 nmol/l) and a calcium ionophore (A23187, 40 mumol/l) but the responses to these secretagogues differed markedly from those to LRH for the tachyphylaxis which resulted from repeated exposure was not preceded by an increase in responsiveness. The decreased responsiveness to K+ developed in parallel with that to LRH. Diminished responses to the cyclic nucleotides and the Ca++ ionophore developed more rapidly, but the refractory cells responded readily to stimulation with LRH or K+.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3024441 TI - End-organ resistance to PTH infusion in hypercalcaemic and normocalcaemic patients with solid tumours. AB - Ten hypercalcaemic patients with solid tumours were studied to evaluate the renal response on PTH infusion as assessed by nephrogenous cAMP excretion and maximum tubular re-absorption of phosphate. In addition, 20 normocalcaemic patients, 11 with an adenocarcinoma and 9 with a squamous cell carcinoma, were studied. All cancer patients had moderately extensive disease. Results were compared with those of 9 patients with primary hyperparathyroidism and with 10 elderly controls. All groups studied had comparable renal function, magnesium and 25 hydroxy-vitamin D levels. Comparable results were obtained in patients with an adenocarcinoma and in controls. cAMP response (delta nephrogenous cAMP) was significantly lower in the hypercalcaemic patients with a solid tumour compared with the controls (8.13 +/- 4.68 nmol/100 ml glomerular filtrate vs 29.52 +/- 25.62 nmol/100 ml glomerular filtrate; P less than 0.005). In the group of patients with primary hyperparathyroidism delta nephrogenous cAMP was 13.41 +/- 7.54 nmol/100 ml glomerular filtrate (P less than 0.06 vs controls). The group of patients with a squamous cell cancer showed an intermediate value of 14.83 +/- 10.74 nmol/100 ml glomerular filtrate (P less than 0.025 vs the normocalcaemic adenocarcinoma patients, but NS vs controls). In two hypercalcaemic patients with a solid tumour in whom PTH infusion was repeated after normalization of serum calcium no influence on renal responsiveness was observed. Responses of maximum tubular re-absorption of phosphate were lowest in the group of hypercalcaemic patients with a solid tumour and in the patients with primary hyperparathyroidism compared with controls (0.11 +/- 0.10 vs 0.22 +/- 0.09 mmol/l and 0.09 +/- vs 0.22 +/- 0.09 mmol/l; P less than 0.025 and P less than 0.005, respectively).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3024442 TI - Growth promoting effect of head activator in cultured chick embryo brain cells. AB - To investigate the regulatory role of head activator (HA) and its synthetic analogue, (Arg1,Phe5)-HA(AHA) on brain cell growth, we measured serial uptakes of [3H]thymidine, [3H]uridine and [3H]leucine and changes in cyclic AMP content in cultured chick embryo brain cells. HA stimulated all of these uptakes at a concentration of 10(-10) M, while 10(-9) M AHA suppressed them. The stimulatory effect of HA on [3H]thymidine uptake was observed after 4 h of the treatment, reached a maximum of 200% of the initial value at 8 h and declined to the pre treatment level at 14 h. [3H]uridine uptake began to increase after 6 h of HA treatment, and the effect lasted for 4 h. Increase in [3H]leucine followed after 12 h and sustained for 4 h. Prior to the initiation of HA stimulation, cyclic AMP also began to rise, reaching 170% of the pre-treatment level at 6 h. In contrast, depression of [3H]thymidine uptake by AHA was noted at 6 h and continued for 8 h. Uptake of [3H]uridine and [3H]leucine showed similar tendency. Cyclic AMP in AHA treated cells at 6 h was significantly lower than that in non-treated cells. These results indicate that HA stimulates DNA, RNA and protein synthesis in an early stage of growing brain cells, in which cAMP may be involved as a regulator of nerve cell growth. PMID- 3024443 TI - Modification of corticosteroid synthesis by etomidate/fentanyl and air anesthesia. AB - The characteristics of cortisol synthesis blockade by an etomidate/fentanyl combination was explored in a group of seven patients undergoing major abdominal vascular surgery. Cortisol, androstene-dione 11-deoxy cortisol (compound S) and ACTH were measured during surgery for three hours. In the fourth hour an ACTH1-24 stimulation test was performed and the reaction of the corticosteroid synthesis was assessed. ACTH and androstene-dione showed a stable concentration during the study, the reaction of androstene-dione to ACTH1-24 was blunted but normal. Compound S and cortisol concentrations decreased during anesthesia and showed no significant increase after stimulation with ACTH1-24. These results indicate that the infusion of etomidate and fentanyl may cause a blockade of the corticosteroid synthesis. The blockade is situated at the place where the conversion of cholesterol to pregnenolone occurs. Because the study was done in clinical setting the results should be interpreted carefully, and confirmed by experimental laboratory results. PMID- 3024444 TI - Bilateral brainstem connections of the rat supratrigeminal region. AB - Efferent and afferent connections of the supratrigeminal region were studied in the rat using iontophoretically delivered horseradish peroxidase and Phaseolus vulgaris leuco-agglutinin. Projections of supratrigeminal efferents were found to the contralateral supratrigeminal region, to the ipsi- and contralateral trigeminal motor nuclei and the medullary reticular formation, and to the ipsilateral facial and hypoglossal motor nuclei. Neurons projecting to the supratrigeminal region were located in the contralateral supratrigeminal nucleus, in the ipsilateral mesencephalic trigeminal nucleus and bilaterally in the medullary reticular formation. This organization is discussed with respect to bilateral oral motor control mechanisms. PMID- 3024445 TI - Arrangement and connections of mesencephalic trigeminal neurons in the rat. AB - The morphology of the mesencephalic trigeminal nucleus was examined microscopically in serial frozen sections. The nucleus extends over a length of about 4.5 mm, and its cell number was calculated to range from 1,000 to 1,600. 60% of the cells were located in the caudal third of the nucleus. Clustering of large unipolar cells was seen throughout the nucleus. Small spindle-shaped multipolar cells were found in the pontine part of the nucleus. The efferent connections of the mesencephalic trigeminal neurons were investigated by means of iontophoretically delivered Phaseolus vulgaris leuco-agglutinin or horseradish peroxidase after electrophysiological identification of mesencephalic trigeminal neurons. All projections were found ipsilateral to the injection site; they were confined to the trigeminal motor nucleus, especially to its lateral part, and to the dorsolateral reticular formation. The latter projection area included the supratrigeminal nucleus, the nucleus of Probst, and the parvocellular reticular zone. There were no direct projections to the facial or hypoglossal motor nuclei. It is concluded that proprioceptive input from one side is mediated polysynaptically to the bilateral oral final common-path neurons, with the exception of the ipsilateral trigeminal motoneurons. PMID- 3024446 TI - Immunoreactivity of surfactant-apoprotein in adenocarcinomas, large cell and small cell carcinomas of the lung. AB - Fifty seven adenocarcinomas, 43 large cell and 30 small cell lung carcinomas were immunohistochemically examined using monospecific IgG against pulmonary surfactant apoprotein. Six peripheral adenocarcinomas and one peripheral large cell carcinoma were found to be histogenetically related to type II pneumocytes. They showed an acinar, papillary or solid growth pattern. The immunohistochemical study using anti-surfactant apoprotein IgG gave a granular reaction product in both the cytoplasm and nucleus of tumor cells whose intensity was variable. Intranuclear inclusions were generally the most densely stained structures. These results suggest that lung carcinomas derived from alveolar type II cells are found not only in bronchioloalveolar tumors, but are also found in other types of adenocarcinoma and in large cell carcinomas. PMID- 3024447 TI - Studies on the pathogenesis of hepatocellular carcinoma in HBV-negative alcoholic cirrhotics. AB - Ninety five cases of HBV marker-negative cirrhosis with excess alcohol intake were examined clinicopathologically to obtain some clues and insights into the pathogenesis of hepatocellular carcinoma (HCC). The following data were obtained: cases were divided morphologically into 37 cases of macronodular cirrhosis (MacCir), 16 mixed cirrhosis (MixCir), and 42 micronodular cirrhosis (MicCir), the mean age at death was the oldest in MacCir (61 yrs), the youngest in MicCir (51 yrs), and intermediate in MixCir (59 yrs), association of HCC was common both in MacCir and MixCir (78 and 63%, respectively) but infrequent in MicCir (17%), all livers of MicCir with HCC had broad collapse and a small number of macronodules in non-cancerous areas and the mean age of them was older than that of MicCir without both the collapse and macronodules (56 vs 48 yrs), in total cases, the mean age at death of patients with HCC was 7 years older than that without HCC (60 vs 53 yrs), the mean liver weight was the largest in MicCir (1,211 g), the smallest in MacCir (829 g), and intermediate in MixCir (1,022 g), the incidence of MacCir was significantly higher in patients who had given up alcohol for more than one year before death than those without abstinence, and neither the subtypes of cirrhosis nor the incidence of HCC was significantly related to the total amount of alcohol intake. These data indicate that the development of HCC in HBV-negative alcoholics with cirrhosis occurs in relation to the development of macronodules and loss of liver weight, most likely along with the prolongation of the life span. PMID- 3024448 TI - Establishment of monoclonal antibodies reactive with epithelial and myoepithelial cells in the breast. AB - Monoclonal antibodies which are considered to be able to differentiate epithelial and myoepithelial cells in the breast have been developed. Human mammary carcinoma cell line (HBC-4W) was used for immunization. Monoclonal antibodies B4B2F10 (epi-1), E9E8B7 (myo-1)-with IgM was examined using tissues from diseased breast by avidin-biotin-peroxidase assay. Epi-1 antibody reacted with epithelial cells while myo-1 antibody reacted with myoepithelial cells in the mammary glands, respectively. The reaction was markedly visible, in particular, in fibroadenoma, mastopathy, and papilloma, which showed clear two-cell-type structures. In the infiltrating ductal carcinoma, epi-1 antibody reacted with carcinoma cells, while myo-1 antibody reacted with stromal cells rather than carcinoma cells suggesting that infiltrating ductal carcinoma was mainly of epithelial origin. In the infiltrating lobular carcinoma, however, myo-1 as well as epi-1 antibodies reacted with carcinoma cells. It is suggested that infiltrating lobular carcinoma was of a mixture of epithelial and myoepithelial cell origin. PMID- 3024449 TI - Restricted heterogeneity of the early antibody response to Aleutian disease virus in mink kits. AB - Mink kits born of Aleutian disease (AD) negative dams were infected neonatally with different isolates of Aleutian disease virus (ADV). They were then sacrificed at different days after infection (10-45 days) and their sera were analysed by different techniques for concentrations of immunoglobulins and quality of the produced antibodies to ADV. The infected mink kits produced significantly higher concentrations of IgM and IgG immunoglobulins than non infected-age matched control mink (p less than 0.001). Eight percent of the sera from the infected mink kits exhibited a restricted heterogeneous gammaglobulin profile as shown by serum electrophoresis. These findings were further investigated by other techniques such as SDS-PAGE and crossed serum line immunoelectrophoresis. It is concluded that mink kits when infected neonatally with ADV start to develop hypergammaglobulinemia soon after infection, and that a small percentage of the mink react with a gammaglobulin response of restricted heterogeneity. PMID- 3024450 TI - Serological diagnosis of mumps and parainfluenza type-1 virus infections by enzyme immunoassay, with a comparison of two different approaches for detection of mumps IgG antibodies. AB - Two solid-phase enzyme-linked immunosorbent assays (ELISA) for detection of mumps IgG antibodies, viz., indirect ELISA and catching-antibodies (C.A.)-ELISA, are described and the results obtained with both assays are compared with each other and with the conventional complement-fixation (CF) test. In the indirect method, mumps antigens are used for coating the wells of the microtest plates, whereas in the C.A.-ELISA method mumps antigens are selectively bound to rabbit anti-mumps antibodies coated surfaces. A positive correlation was found between the optical density (O.D.) values given by both ELISA assays and CF-antibody titers. The ELISA assays showed improved sensitivity compared to the CF test, since 54% (C.A. ELISA) and 33% (Indirect-ELISA) of additional positive reactions were detected by these assays. In terms of specificity, however, only the C.A.-ELISA was superior to CF, since significant rises of IgG-antibodies were detected only in paired sera of mumps patients. Conversely, when the indirect method was used significant IgG antibody rises were demonstrated in paired sera from mumps patients and in serum pairs of six patients with parainfluenza type-1 virus infection. With the CF test, heterologous antibodies responses were demonstrated in 2 of these patients. Absorption experiments of mumps sera with mumps and parainfluenza virus strains demonstrated that the IgG antibodies detected by the C.A.-ELISA are specific for mumps virus and therefore interference due to heterologous antibody responses were not observed. Results with purified mumps virus proteins demonstrated that the antigen-antibody reactions that partake in the C.A.-ELISA are mainly associated with the nucleoprotein antigen. Detection of IgG-antibodies to parainfluenza virus type-1 was assessed by ELISA (paraflu-T1-ELISA) using only the indirect approach. The results obtained with this assay showed improved sensitivity compared to a paraflu T1-CF test, since 47% of additional positive reactions were demonstrated by ELISA. In terms of specificity, however, heterologous antibody responses were detected by both the ELISA and the CF test in 4 out of 20 patients with mumps infections. PMID- 3024451 TI - [Chromatography of Bungarus fasciatus venom and preliminary studies of its neurotoxin--fraction IX]. PMID- 3024452 TI - [Effects of polymyxin B on action potentials of mice right ventricular myocardial cell]. PMID- 3024453 TI - Alveolar lymphocyte proliferation induced by Propionibacterium acnes in sarcoidosis patients. AB - The proliferation of lymphocytes induced by Propionibacterium acnes (P. acnes) was measured by the in vitro incorporation of 3H-thymidine. The mean response rate of alveolar lymphocytes obtained by bronchoalveolar lavage was 2.23 +/- 0.89 in nine untreated sarcoidosis patients, 0.85 +/- 0.17 in five sarcoidosis patients given corticosteroids and 0.78 +/- 0.29 in 11 controls. The proliferation was significantly enhanced in the untreated patients compared to both the treated patients (p less than 0.01) and controls (p less than 0.001), but there was no significant difference in response rates between the treated patients and controls. The response rate of alveolar lymphocytes was significantly higher in four active patients (3.05 +/- 0.61) than in four inactive patients (1.77 +/- 0.44) (p less than 0.05) and in the controls (p less than 0.001). In sarcoidosis patients, the response rates showed a good correlation with activities of serum lysozyme (r = 0.695, p less than 0.01), and with percentages of lymphocytes in bronchoalveolar lavage fluid (r = 0.591, p less than 0.05). There was a low correlation between angiotensin-converting enzyme activities and the response rates (r = 0.508, p less than 0.1). Neither peripheral blood lymphocytes in sarcoidosis patients nor in controls showed any response to P. acnes, but alveolar lymphocytes of the untreated active sarcoidosis patients were sensitive to P. acnes. The lymphocytes activated by P. acnes may play a central role in the induction of alveolitis in sarcoidosis patients. PMID- 3024454 TI - Laboratory diagnosis of herpes viruses in the immunocompromised host. PMID- 3024455 TI - Herpesvirus infections in the immunocompromised host: diagnosis and management. PMID- 3024456 TI - Anatomical organization of excitatory amino acid receptors and their properties. PMID- 3024457 TI - Na+ fluxes as a tool to identify anticonvulsant antagonists of neuroexcitation. PMID- 3024458 TI - Involvement of excitatory amino acid receptors in the mechanisms underlying excitotoxic phenomena. PMID- 3024459 TI - Excitatory amino acid antagonists as novel anticonvulsants. PMID- 3024460 TI - Expression of vertebrate amino acid receptors in Xenopus oocytes. PMID- 3024462 TI - Excitatory amino acids and divalent cations in the kindling model of epilepsy. PMID- 3024461 TI - Neurobehavioral, neuroendocrine and neurochemical effects of zinc supplementation in rats. PMID- 3024463 TI - Effect of zinc on neuronal activity in the rat forebrain. AB - Zinc ions, which are unevenly distributed in the CNS and can be released from nerve terminals, have been implicated as causative agents in epileptogenesis. The present study has shown that intraventricular administration to anesthetized rats causes seizure activity of the ECOG and convulsions. Since the manner in which zinc influences neuronal activity and triggers convulsions is unclear, studies were also made of its effect on spontaneous and evoked activity in the rat forebrain. It was found that iontophoretic application of zinc to cortical neurons causes slow and often prolonged increases in firing rate, usually accompanied by bursts of high frequency discharge in just under half the studies. Another cation, barium, evoked excitatory responses of a similar type and a reduction in potassium permeability may underlie the effects of both cations. In contrast, calcium, magnesium, manganese and cerium caused short duration depressant effects. The depression induced by calcium, but not by the other cations, could be blocked by zinc. Similarly, in the hippocampus zinc depressed calcium-dependent potentiation in subfield CA3 evoked by paired-pulse stimulation of mossy fibers; excitatory effects (namely an increase in spike amplitude and appearance of multiple population spikes) were seen at higher zinc concentrations. The depressant effects of an enkephalin analog on cortical firing rate were also blocked by zinc, consistent with studies from another laboratory suggesting enkephalin/zinc interactions. In contrast, the depressant effect of GABA could not be blocked by zinc, although an antagonism has been reported in the lobster muscle. Firm conclusions regarding the mechanism(s) underlying the triggering of seizure activity by zinc cannot yet be drawn, but the results of these studies would be consistent with an interference with calcium and/or potassium ion activity rather than with GABA binding sites. PMID- 3024465 TI - Selective and non-selective seizure related brain damage produced by kainic acid. PMID- 3024464 TI - Inciting excitotoxic cytocide among central neurons. AB - Here I have reviewed evidence from electron microscopic studies showing that each of several sustained limbic seizure syndromes is associated with a type of acute brain damage which is ultrastructurally indistinguishable from the brain damage induced by Glu and other excitotoxins. In addition, I have presented evidence that persistent stimulation of specific axonal tracts that use Glu as transmitter results in Glu-like excitotoxic degeneration of postsynaptic neurons innervated by such tracts. Phencyclidine and ketamine, which powerfully block the neurotoxicity of the Glu analog NMA, protect against seizure-related brain damage. This may be explained by either an anticonvulsant or antiexcitotoxic mechanism, or both. Recent evidence suggests that an excitotoxic mechanism (excessive activation of Glu/Asp receptors) may underlie both seizure-mediated and anoxic brain damage. The acute fulminating type of neuronal degeneration induced by Glu is a Na+ and Cl- but not Ca2+ dependent phenomenon. According to a recent study, however, Glu may induce neuronal necrosis not only by an acute Ca2+ independent process but by a more slowly evolving Ca2+ dependent process. If, as these data suggest, an excitotoxic mechanism underlies brain damage associated with anoxia and epilepsy, a better understanding of excitotoxic mechanisms may lead eventually to prophylactic approaches for preventing such forms of brain damage. PMID- 3024466 TI - Quinolinic acid: a pathogen in seizure disorders? AB - The evidence for an involvement of QUIN in human seizure disorders is clearly circumstantial. Importantly, QUIN is not a classical neurotransmitter and may thus play only a negligible or no role at all in normal brain function (Foster et al., 1984). We have yet to understand if and how such a possibly inert metabolite may turn into a pathogen. Several crucial questions remain to be addressed before a case can be made for a 'quinolinic acid hypothesis' of temporal lobe epilepsy. Among the most prominent ones figure the extracellular concentration of QUIN in the human brain under normal and pathological ('epileptic') conditions, the relationship between QUIN metabolism in the brain and its extracellular concentration and, a related issue, the regulation of cerebral QUIN metabolism (i.e., turnover). It is of equal importance to assess if NMDA-receptors, particularly those in the hippocampus and other parts of the limbic system, can exert a modulatory function upon brain QUIN. Unquestionably, future experiments with selective NMDA-antagonists will prove useful for the elucidation of such possible (feedback) interactions. PMID- 3024467 TI - Functional adaptations of transbilayer proteins. PMID- 3024468 TI - The interaction of egg peptides with spermatozoa. PMID- 3024469 TI - Internalization of hormone receptor complexes: route and significance. PMID- 3024470 TI - Ligand-receptor dynamics and signal amplification in the neutrophil. PMID- 3024471 TI - [A case of metastatic urinary bladder tumor from a gastric remnant carcinoma]. AB - We report a case of a metastatic urinary bladder tumor from gastric remnant carcinoma. On August 23, 1984, a 70-year-old-woman visited us with the complaint of dysuria. She had undergone gastrectomy for gastric ulcer 25 years earlier. Cystoscopy revealed a non-stalk tumor in the dome of the bladder and the examination of the upper gastro-intestinal tract revealed gastric remnant carcinoma. We treated her with adriamycin, cis-diamminedichloroplatinum and mitomycin C but unfortunately she died of cachexia two weeks later. An autopsy revealed that the urinary bladder tumor was a signet ring cell carcinoma, metastasized from gastric remnant carcinoma. PMID- 3024472 TI - Malignant gestational trophoblastic disease: CT findings. AB - Eight patients with malignant gestational trophoblastic disease had CT of the pelvis as part of their staging before chemotherapy. CT appearance of the uterus fell into three major types: normal size with irregular areas of hypodensity, uniform enlargement with areas of hypodensity, and focal areas of enlargement with or without areas of hypodensity. Patients with the second and third types were much more likely to have distant metastases and to require hysterectomy for successful treatment. CT findings of tumor nodules in the parametrium or myometrium were confirmed in three of the four surgical specimens of the uterus available for correlation. Myometrial tumor nodules were seen as areas of focal enlargement or as irregular eccentric areas of hypodensity. In one patient, parametrial extension was seen as an enhancing mass adjacent to the uterus. CT may be accurate in defining the extent of myometrial and adnexal disease and may have prognostic and therapeutic value. PMID- 3024473 TI - MR imaging of paragangliomas. AB - MR imaging of 15 paragangliomas in 10 patients was compared with CT of 13 of the lesions in eight patients. All lesions were confirmed with angiography. All lesions were detected by MR and CT with the exception of one small glomus tympanicum tumor that was seen only in retrospect with MR. CT better demonstrated subtle osseous changes of the skull base and the relation of the tumor to the middle ear structures. MR better demonstrated the relation of the tumor to the adjacent internal jugular vein and carotid artery. The paragangliomas had a characteristic MR appearance based on their vascularity. Serpiginous areas of signal void representing high vascular flow were interspersed among areas of high signal intensity caused by slowly flowing blood and tumor cells. This "salt-and pepper" pattern was seen in all lesions greater than 2 cm in maximal dimension. MR was therefore able to accurately characterize the tumors as highly vascular. Multiplanar imaging and good tissue contrast and anatomic detail permitted display of the relations of these neoplasms to surrounding carotid sheath vessels and to intracranial structures better than did CT. In this experience, the MR appearance of paragangliomas was quite characteristic and differed markedly from meningiomas, neuromas, and metastatic disease of the skull base. PMID- 3024474 TI - Embolization of epistaxis and juvenile nasopharyngeal angiofibromas. AB - This article reviews arterial embolization of epistaxis and juvenile nasopharyngeal angiofibromas. A protocol for the complete and rapid exploration of epistaxis is suggested that is based upon which arteries providing blood to the nasal fossa may be responsible for the bleeding and the most optimal way for their demonstration and evaluation. The relevant anatomy of the major arteries, their branches, and important anastomoses are described. The goals of embolization are to control severe or recurrent epistaxis, prevent recurrence if possible, and avoid occlusion of any vessels not responsible for the hemorrhage. The most appropriate embolic material is chosen, realizing that recurrence can be caused by the use of resorbable embolic material or by a very proximal or too complete embolization. Embolization may be used for specific causes of epistaxis including idiopathic, hereditary hemorraghic telangiectasia, hemorrhagic tumors, vascular malformations, trauma, disorders of hemostasis, and postsurgical problems. Juvenile nasopharyngeal angiofibromas are discussed with respect to their clinical presentation, classification, computed tomographic, MR imaging, and angiographic evaluation, treatment, future trends in treatment, and precautions and complications. PMID- 3024476 TI - Cardiac alpha-receptors and cardiac hypertrophy in genetic predisposition to hypertension. PMID- 3024475 TI - Diagnosis of subclinical varicocele in infertility. AB - The clinically obvious varicocele is perhaps the most common identifiable and correctable cause of male infertility. However, less is known about the subclinical (not palpable) varicocele and its relationship to infertility. We undertook this study to compare the ability of high-resolution sonography and radionuclide scrotal scanning to detect subclinical varicocele. Fifty patients who were referred to our department with a diagnosis of infertility, an abnormal semen analysis, and a normal physical examination of the scrotum underwent both sonography and nuclear scanning. The final study group included 20 men who agreed to surgical ligation of the spermatic vein(s) after a positive sonographic and/or radionuclide study. Sonography was considered positive for subclinical varicocele in 95% of patients, while nuclear scanning was considered positive in only 55%. Postoperatively, all patients showed improvement in their semen and 40% (eight patients) became fertile. Subclinical varicocele seems to be an important causal factor in infertility and, in our experience, high-resolution sonography is superior to radionuclide scanning in its diagnosis. PMID- 3024477 TI - Relationship of dietary fat, protein, cholesterol, and fiber intake to atherogenic lipoproteins in men. AB - Nutritional components (g/1000 kcal) obtained from 3-day diet records are compared to triglyceride, total cholesterol, low-density (LDL), intermediate density (IDL), and very low-density (VLDL) lipoprotein concentrations of 77 free living men. Polyunsaturated-fatty acid consumption correlated negatively with concentrations of triglycerides, total cholesterol, LDL- and VLDL-cholesterol, and total-lipoprotein mass of smaller-LDL particles (Sf0 0-7), IDL (Sf0 12-20), and VLDL (Sf0 20-400) in serum and plasma. Animal-protein consumption correlated positively and plant-protein consumption correlated negatively with triglycerides, smaller-LDL mass, VLDL-cholesterol, and VLDL-mass levels. Serum concentrations of smaller-LDL particles were also positively correlated with dietary-cholesterol intake and negatively correlated with crude-fiber consumption. Thus, diet-lipoprotein relationships observed cross-culturally and experimentally are further supported when detailed dietary measurements from 3 day diet records and lipoprotein measurements from repeated blood samplings are correlated in free-living men. PMID- 3024478 TI - The relationship between estrogen levels and diets of Caucasian American and Oriental immigrant women. AB - The relationship between diet and estrogens was studied in two groups of women with different dietary habits and breast cancer risks. Plasma estrogens and androgens and 24-h urinary and fecal excretion of estrogens were measured in premenopausal and postmenopausal Caucasians and recent Oriental immigrants from Southeast Asia to Hawaii. Premenopausal Caucasians had 30-75% higher plasma estrone and estradiol levels than their age-matched cohorts in Hawaii, and the postmenopausal Caucasians had 3-fold higher plasma levels of estradiol. The Oriental women excreted more than twice the amount of estrogen in their feces but they excreted significantly less in their urine. Thus, the ratio of urinary-to fecal excretion was approximately 3-5 times higher in young Caucasian women. Analysis of dietary components and plasma estrogens in premenopausal women showed a positive correlation between daily intake of total fat and saturated fat and plasma estrone and estradiol concentrations. PMID- 3024479 TI - Inflammatory breast carcinoma. PMID- 3024480 TI - False positive indirect fluorescent antibody (IFA) tests for antibody to herpes simplex virus 1. Comparison of four commercially available methods. AB - Four commercially available herpes simplex virus 1 (HSV-1) indirect fluorescent antibody (IFA) kits were compared with the use of sera selected because they were negative for HSV antibody by complement fixation (CF less than 1:8) and by ELISA (less than 1:100). However, 14 of 24 (58.3%) of these HSV-1 antibody-negative sera were positive at greater than or equal to 1:10 with the use of the HSV-1 IFA kit from Electronucleonics, 15 of 24 (62.5%) were positive with the Clinical Sciences HSV-1 IFA kit, 4 of 24 (16.7%) were positive with Zeus Scientific, and 4 of 18 (22.2%) were positive with the Gull Laboratories product. HSV-1 induces Fc receptors that commonly cause false positive IFA tests for HSV antibody. Therefore, further studies were undertaken to determine whether Fc receptors accounted for these false positive results. Staphylococcal protein A (SPA) is known to bind to the Fc portion of human IgG and therefore could be used to distinguish between the binding of an antibody by its Fab or its Fc portion. Thus, when fluorescein isothiocyanate conjugated (FITC) SPA was used as conjugate instead of FITC antibody to human IgG, true HSV-1 antibody-containing sera remained positive, but the false positives identified in the commercial IFA kits did not. The authors conclude that HSV-1-induced Fc receptors are responsible for most of the problem of these false positives and that HSV-1 serology probably should not be done by this type of IFA method until this problem is corrected. PMID- 3024481 TI - Hepatic disorders in the acquired immune deficiency syndrome: a clinical and pathological study. AB - We reviewed the clinical data, hepatic histology, and microbiological features of 21 patients with previously documented acquired immune deficiency syndrome who had liver biopsies. Diagnoses of specific infections were made on liver biopsy in 11/21 patients (57%). Granulomas were found in 10/21 patients (48%) and were most often a manifestation of infection with Mycobacterium avium-intracellulare. Elevated levels of serum alkaline phosphatase and longer duration of diagnosed illness were significantly associated with the presence of granulomatous disease. PMID- 3024482 TI - Mortality from lung cancer and respiratory disease among pottery workers exposed to silica and talc. AB - A cohort mortality study of white men employed for at least one year between 1939 and 1966 at three plants of a single United States company was conducted to evaluate the risk of lung cancer and nonmalignant respiratory disease among workers exposed to silica dust and nonfibrous (nonasbestiform) talc in the manufacture of ceramic plumbing fixtures. Follow-up of 2,055 men through January 1, 1981, indicated a substantial excess of nonmalignant respiratory disease among those with high levels of exposure to silica dust (standardized mortality ratio = 2.26). The risk of nonmalignant respiratory disease rose with the number of years exposed, was not further enhanced by talc exposure, and appeared to be appreciably lower among those exposed in more recent time periods. For lung cancer, men exposed to high levels of silica dust with no talc exposure had a nonsignificant standardized mortality ratio of 1.37. However, those exposed to nonfibrous talc in addition to high levels of silica had a significant 2.5-fold excess risk of lung cancer. Among this group, the lung cancer standardized mortality ratio rose with increasing years of talc exposure to 3.64 among those exposed for 15 or more years. Although the role of silica as a cofactor cannot be ruled out, these data suggest that nonfibrous talc exposure is associated with excess lung cancer risk. PMID- 3024483 TI - Segregation of marker loci in families with an inherited paracentric insertion of chromosome 9. AB - Cytogenetic, enzyme dosage, serological, and electrophoretic analyses of blood samples from members of three Newfoundland kindreds in which one specific paracentric insertion chromosome inv ins(9)(q22.1q34.3q34.1) is segregating provide data indicating that ABO lies in 9q22.1-q34.3, AK1 in 9q34.1-q34.3, and ORM in 9q34.3-qter. PMID- 3024484 TI - Early detection and signs of hepatoangiosarcoma among vinyl chloride workers. AB - Health examinations of 108 workers exposed to vinyl chloride monomer (VCM) at a Japanese chemical plant were carried out in 1979. The polymerization of vinyl chloride was started at the plant in 1949. In this study, the highest concentration of VCM in autoclaves was determined to be 250 ppm in 1961. However, the workers at the plant had been exposed to higher concentrations of VCM several times before 1960. More recent VCM exposure was considered negligible. Examinations assessed data on age, height, weight, obesity index, sake consumption, VCM exposure concentration, latent period, cumulative exposure, ICG (indocyano green test), serum bilirubin, GOT (glutamic oxaloacetic transaminase), GPT (glutamic pyruvic transaminase), A1-P (alkaline phosphatase), GGT(gamma glutamyl transpeptidase), ZTT (zinc turbidity test), LDH (lactate dehydrogenase), cholesterol, TTT (thymol turbidity test), A/G (albumin globulin ratio), and thrombocytes. Variation in VCM exposure did not affect tests of pigment excretion from the liver, such as ICG; thrombocytes; and enzyme activity (such as GPT); nor bilirubin or flocculation reaction in serum. PMID- 3024486 TI - Perlman syndrome: familial renal dysplasia with Wilms tumor, fetal gigantism, and multiple congenital anomalies. PMID- 3024485 TI - Familial growth hormone deficiency resulting from a 7.6 kb deletion within the growth hormone gene cluster. AB - We report on two sibs with familial isolated growth hormone deficiency (IGHD) resulting from homozygosity for a 7.6 kb deletion within the growth hormone gene cluster. The deletion not only affects the structural gene for growth hormone (GH N) but also alters sequences adjacent to the chorionic somatomammotropin-like (CS L) gene. In contrast to previously reported cases with IGHD type IA, our two patients responded well to growth hormone substitution and formation of blocking antibodies did not occur. PMID- 3024487 TI - Forskolin: unique diterpene activator of adenylate cyclase in pregnant and nonpregnant guinea pig myometrial membranes. AB - In guinea pig myometrium, beta-adrenergic receptors are functionally coupled to adenylate cyclase. beta-Adrenergic receptor agonists in the presence of guanosine triphosphate stimulate adenylate cyclase activity, thus increasing 3'5'-cyclic adenosine monophosphate synthesis and promoting myometrial relaxation. In pregnant animals close to term (65 days), beta-adrenergic receptor density as well as basal and (-)isoproterenol-dependent (in the presence of guanosine triphosphate) adenylate cyclase activity is significantly higher than that in nonpregnant animals or those in early pregnancy. Since this system appears to be made up of at least three components (beta-adrenergic receptor, guanosine triphosphate-binding protein, and a catalytic component), these observations on total adenylate cyclase activity may reflect alterations in one or more of these components. To answer the question whether the catalytic unit of this system can be directly assayed and whether its activity is influenced by pregnancy, we have performed in vitro experiments to measure the enzymatic activity of the catalytic component of the beta-adrenergic receptor-adenylate cyclase complex in guinea pig myometrial membranes. We have used two compounds that stimulate the catalytic component: forskolin and manganese chloride. Forskolin, regardless of the presence or absence of guanosine triphosphate, is the most potent stimulator of adenylate cyclase activity in myometrial membranes from nonpregnant and pregnant animals; manganese chloride is a less potent activator. The degree of adenylate cyclase stimulation by forskolin tends to be higher in uteri from pregnant (greater than or equal to 0.5 gestation) than from nonpregnant or postpartum animals. It was concluded: that adenylate cyclase stimulation by forskolin does not depend on the presence of beta-adrenergic receptor agonists or guanosine triphosphate and that with advancing gestation there might be a qualitative or quantitative change with regard to the interaction between forskolin and the presumed catalytic component of the beta-adrenergic receptor-adenylate cyclase complex. PMID- 3024488 TI - The effect of estrogen-progestin treatment on opioid control of gonadotropin and prolactin secretion in postmenopausal women. AB - Naloxone (10 mg) was given intravenously to seven postmenopausal women not receiving hormone treatment and to six postmenopausal women receiving Premarin Provera treatment during the Premarin phase and also during the Premarin-Provera phase of therapy. Baseline estrone and estradiol levels (mean +/- SEM) were significantly lower in the group not receiving hormones (46.0 +/- 5.2 pg/ml and 28.4 +/- 3.1 pg/ml, respectively) than in the group in the Premarin phase of therapy (154 +/- 14 pg/ml and 79 +/- 13 pg/ml) and the group in the Premarin Provera phase (135.1 +/- 8.3 pg/ml and 57.5 +/- 3.0 pg/ml) (p less than 0.005). Follicle-stimulating hormone, luteinizing hormone, and prolactin levels were 118.7 +/- 5.3 mIU/ml, 118.7 +/- 9.5 mIU/ml, and 9.2 +/- 0.7 ng/ml, respectively, with no significant change after naloxone administration in untreated women. With hormone therapy the basal follicle-stimulating hormone and luteinizing hormone levels decreased significantly while basal plasma estrone and estradiol increased significantly. In both the group in the Premarin phase of therapy and the group in the Premarin-Provera phase, luteinizing hormone levels increased significantly at 30 (135% +/- 10%, 144% +/- 8%), 45 (150% +/- 12%, 133% +/- 11%), 60 (149% +/- 15%, 128% +/- 11%), and 90 (139% +/- 15%, 132% +/- 13%) minutes after naloxone administration (p less than 0.01 to p less than 0.001). Follicle-stimulating hormone levels did not change significantly whereas prolactin levels showed a trend toward a decrease. These findings indicate that opioid inhibition of gonadotropins is reduced in postmenopausal women but increased with Premarin Provera treatment. The effect of sex steroid on the opioid system in the postmenopausal women differs from that in the premenopausal women. PMID- 3024489 TI - Forskolin-stimulated adenylate cyclase activity in fetal and adult rabbit myocardial membranes. AB - It has previously been reported that there is an increase in the concentration of myocardial beta-adrenergic receptors as well as an increase in the sensitivity of adenylate cyclase stimulation by L-isoproterenol in fetal rabbit myocardial membranes from 21 to 31 days' gestation. To investigate whether a change in the catalytic component of the beta-adrenergic receptor-adenylate cyclase complex is responsible for the observed increase in adenylate cyclase activity, I have measured the latter in the presence of forskolin and manganese chloride, compounds that can directly stimulate the catalytic component. Myocardial membranes were obtained from fetal rabbits at 21, 28, and 31 days' gestation and from pregnant adult animals. There was an increase in basal adenylate cyclase activity from the fetal to the adult stage. The percent of maximal stimulation by L-isoproterenol in fetuses from 21 through 28 to 31 days' gestation and in the adult animal. However, although forskolin was the most potent stimulator of adenylate cyclase, the degree of stimulation was equivalent in all groups tested. Similar results were obtained with manganese chloride. We conclude that the catalytic component of the beta-adrenergic receptor-adenylate cyclase complex in myocardial membranes is stimulated to an equivalent degree at all gestational ages in fetal and adult rabbits and shows no maturational changes. PMID- 3024490 TI - Plasma adrenocorticotropic hormone and cortisol and adrenal blood flow during sustained hypoxemia in fetal sheep. AB - We examined the effect of sustained hypoxemia with progressive acidemia on pituitary-adrenal endocrine function (adrenocorticotropic hormone, cortisol) and on adrenal blood flow in fetal sheep. Hypoxemia was induced by the maternal sheep breathing a gas mixture containing 9% oxygen, with 3% carbon dioxide added. Induced hypoxemia resulted in a progressive fetal metabolic acidosis but with little change in maternal pH. During induced hypoxemia there was little change in maternal plasma adrenocorticotropic hormone or cortisol level. Fetal adrenocorticotropic hormone and cortisol increased to peak values within 2.8 hours of induced hypoxia but by 7.2 hours had begun to fall to values that were not significantly different from those at 1.4 hours. Fetal adrenal blood flow (microsphere technique) also increased significantly and remained elevated throughout the duration (7.2 hours) of hypoxemia. The maximum fetal adrenal blood flow achieved during hypoxemia was significantly correlated with the basal (prehypoxemia) flow to the adrenals. We conclude that the changes in fetal adrenocorticotropic hormone, cortisol, and adrenal blood flow seen in short-term hypoxemia are reproduced during sustained hypoxemia with acidemia. Furthermore, the noted rise in the fetal adrenocorticotropic hormone level may be an important factor contributing to the increase in adrenal blood flow during hypoxemia. PMID- 3024491 TI - Coxsackievirus B-3 myocarditis. T-cell autoimmunity to heart antigens is resistant to cyclosporin-A treatment. AB - A cardiotropic variant of coxsackievirus group B, Type 3 (CVB3) induces myocarditis in inbred Balb/c mice. Myocardial injury is predominantly mediated by T lymphocytes recognizing normal myocyte antigens, making this an autoimmune disease. Nonetheless, the autoimmune response cannot be inhibited by cyclosporin A (CSA) treatment of the infected animals. Mortality in treated mice was increased 2-4 times, but neither virus-specific antibody or cytolytic T lymphocyte responses were affected, and maximal virus concentrations in the hearts of CSA-treated and control animals were similar. Cardiac damage remains T cell-mediated, because mice given both CSA and rabbit anti-thymocyte serum (ATS) failed to develop significant myocardial inflammation. CSA did suppress immunity in Balb/c mice to an allogeneic C57B1 lymphoma, EL4. Subcutaneous inoculation of mice with 2.5 X 10(6) ascites tumor cells resulted in 100% of the CSA-treated animals having tumors averaging 340 mg 10 days later, compared with less than 10% of control animals having tumors averaging only 30 mg. Humoral immunity to the tumor was absent in CSA-treated mice. Therefore, whether CSA induces immunosuppression depends upon the antigenic stimulus used. PMID- 3024492 TI - Presence of neurophysins in the human pituitary corticotrophs, Cushing's adenomas, and growth hormone-producing adenomas detected by immunohistochemical study. AB - Neurophysins have been recognized as the carrier proteins of vasopressin and oxytocin. The distribution of neurophysins is immunohistochemically confirmed in the hypothalamus, median eminence, and posterior lobe of the pituitary gland. The authors detected neurophysins in the human corticotrophs and pituitary adenomas with the use of the immunohistochemical method with antiserum to human neurophysins, which did not cross-react with adrenocorticotropic hormone (ACTH), beta-endorphin, and corticotropin-releasing factor. All of ten pituitary glands obtained by autopsy revealed the presence of neurophysin-positive cells in the anterior, intermediate, and the posterior lobes. The neurophysin-positive cells were similar to the corticotrophs in shape and distribution. Simultaneous staining for ACTH and neurophysins in the serial sections revealed that neurophysin-positive cells were also ACTH-positive. One hundred twenty-four cases of pituitary adenoma operated upon were investigated. All of 7 Cushing's adenomas were composed of neurophysin-positive cells. Six tumors with giantism showed sparsely distributed neurophysin-positive cells. No neurophysin-positive cells were observed in any other cases. This study is the first reported evidence of the presence of neurophysins in the human corticotrophs and pituitary adenomas. PMID- 3024494 TI - Suprascapular neuropathy related to the use of crutches. AB - A 41-year-old man with shoulder pain was found to have a suprascapular neuropathy. The nerve injury was believed to be secondary to improper crutch usage. The pathophysiology of this unusual cause of suprascapular neuropathy involves injury to the nerve due to exaggerated movements around the shoulder joint. Suprascapular entrapment neuropathy is another potential hazard of improper crutch usage. Therefore, properly fitted crutches and adequate crutch training are essential for the prevention of additional disability. PMID- 3024493 TI - Dietary cholesterol-induced changes in macrophage characteristics. Relationship to atherosclerosis. AB - In diet-induced hypercholesterolemia, circulating monocytes adhere to the endothelium of the vessel wall and emigrate into the intima. Atherosclerotic lesions may develop, characterized by the presence of lipid-laden macrophages and proliferating smooth muscle cells recruited from the media. Using rat peritoneal macrophages, the authors examined the influence of diet-induced hypercholesterolemia on several variables of macrophage function that may contribute to lesion formation, including adhesion to bovine aortic endothelial cells (BAECs) and vascular smooth muscle cells (VSMCs), the production of chemoattractants and mitogens for VSMCs, and the release of the reactive oxygen species, superoxide. In general, a hypercholesterolemia-induced augmentation of macrophage function was observed. In comparison with macrophages from normal animals (N M phi s), macrophages from hypercholesterolemic animals (H M phi s) were 50-80% more adhesive to BAECs and VSMCs. H M phi-secreted products increased VSMC migration 6 to 7-fold, whereas N M0s only stimulated motility 2.5-fold. In addition, H M phi-conditioned media produced increased VSMC growth 5-fold, compared with a 2.5-fold increase produced by N M phi-conditioned media. Although the production of superoxide was found to be the same for both N M phi s and H M phi s, the release of superoxide by macrophages found in the intima of hypercholesterolemic animals may contribute to the necrosis of cells in the developing lesion. These results suggest that dietary cholesterol may accelerate atherosclerotic lesion formation by inducing specific changes in the properties of circulating monocytes and intimal macrophages. PMID- 3024495 TI - 3-O-methyl-D-glucose transport in tumoral insulin-producing cells. AB - Tumoral insulin-producing cells of the RINm5F line were exposed at different temperatures, and for various lengths of time to increasing concentrations of 3-O methyl-D-[U-14C]glucose. The uptake of the hexose represented a temperature sensitive and saturable process, so that no rapid equilibration of hexose concentrations across the plasma membrane was reached, especially at low temperature and/or high concentrations of 3-O-methyl-D-glucose. The uptake of 3-O methyl-D-[U-14C]glucose was not affected by a prior loading of the cells with the unlabeled hexose and its release from prelabeled cells was observed in the absence of any concentration gradient across the plasma membrane. The uptake of D [U-14C]glucose and utilization of D-[5-3H]glucose was inhibited by 3-O-methyl-D glucose, which failed, however, to affect D-[U-14C]glucose oxidation. At variance with the situation found in normal insulin-producing cells, the transport of D glucose into the tumoral cells may thus play a regulatory role in its metabolism. PMID- 3024496 TI - D-glucose transport and concentration in tumoral insulin-producing cells. AB - The uptake of D-[U-14C]glucose and D-[5-3H]glucose by tumoral insulin-producing cells of the RINm5F line was measured over 3-30 min of incubation at 7-37 degrees C. The apparent distribution space of the hexose ranged from values as low as the L-[1-14C]glucose space to values 10 times higher than the 3H2O space. Although a major fraction of the radioactivity recovered in the cellular pellet corresponded to the metabolites of D-glucose, the results suggested that the transport of D glucose into the tumoral cells represents a saturable and temperature-dependent process. When D-glucose was measured by an enzymic procedure in cells incubated for 30 min at 37 degrees C in the presence of 50.0 mM D-glucose, the apparent distribution space of the hexose remained lower than the intracellular water space. These results indicate that the RINm5F cells have lost an essential attribute of the glucose-sensing device in normal insulin-producing cells, namely the ability to ensure the equilibration of D-glucose concentrations across the plasma membrane. PMID- 3024497 TI - Na+-H+ exchanger of human placental brush-border membrane: identification and characterization. AB - Syncytiotrophoblast brush-border membrane vesicles isolated from full-term human placentas were shown to transport Na+ against a concentration gradient in the presence of an outward proton gradient [( H+i] greater than [H+]o). This proton gradient-coupled Na+ uptake was markedly inhibited and the uphill transport abolished when the electrochemical proton gradient was dissipated by carbonylcyanide 4-(trifluoromethoxy) phenylhydrazone. The presence of nigericin also eliminated the concentrative uptake of Na+ in these vesicles. Dimethylamiloride and harmaline inhibited the proton gradient-induced Na+ uptake. The apparent inhibition constant for this process was 0.32 microM for dimethylamiloride was freely reversible and the inhibitor reduced the Na+ uptake by directly interacting with the exchanger protein rather than by dissipating the H+ gradient. The dimethylamiloride-sensitive Na+ uptake was saturable with respect to Na+. The affinity constant for Na+ was 7.8 +/- 1.2 mM and the maximal velocity was 38.7 +/- 2.4 nmol X mg protein-1 X min-1. The dimethylamiloride insensitive Na+ uptake was not saturable and probably represented simple diffusion. The diffusional component accounted for only 10% of the total uptake. Li+ strongly competed with Na+ for the uptake process and the apparent inhibition constant was 3.6 +/- 0.4 mM. Tetraethylammonium also caused significant inhibition of Na+ uptake, whereas K+, Rb+, Cs+, and choline had no effect. These data provide evidence for the existence of a Na+-H+ exchanger in human placental brush-border membrane, and the properties of this exchanger are similar to those of the Na+-H+ exchanger identified in the brush-border membrane of mammalian kidney and small intestine.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3024498 TI - Calcium binding to cardiac sarcolemmal vesicles: potential role as a modifier of contraction. AB - Passive Ca binding to cardiac sarcolemmal vesicles isolated from rabbit ventricles was measured under ionic conditions similar to intracellular and extracellular media. The first of two main goals was to evaluate whether certain agents induce changes in Ca binding at the external sarcolemmal surface that might contribute to the overall effect of these agents on cardiac muscle contraction. The agents studied were ouabain, verapamil, nifedipine, Bay K 8644, caffeine, ryanodine, and milrinone over a broad range of concentrations, including concentrations at which these agents exert strong effects on cardiac contractile performance. None of these agents produced significant alterations in Ca binding, such that it is unlikely that any part of their actions can be attributed to changes in Ca binding to the external sarcolemmal surface. In contrast, when [Na] is reduced from 140 mM, sarcolemmal Ca binding increases or decreases depending on what replacement is used to avoid changes of osmolarity. Thus the possible effect of Na reduction on surface Ca must be considered in physiological experiments where extracellular [Na] is changed. The second main goal was to evaluate the effects of membrane potential, Na and Mg on Ca bound to the inner surface of the sarcolemma under ionic conditions similar to those expected intracellularly (e.g., [Ca] = 0.3-5.0 microM). Ca binding was inhibited by physiological concentrations of Na and Mg and was sensitive to membrane potential such that depolarization of a normally polarized cell would cause Ca to be released from these sarcolemmal sites. From a quantitative standpoint, it is not clear whether the effect of depolarization would be to contribute sarcolemmal Ca to the activation of the myofilaments or merely to limit the ability of the inner sarcolemmal surface to buffer the rise in intracellular [Ca] associated with contraction. PMID- 3024499 TI - Coordinate regulation of zinc metabolism and metallothionein gene expression in rats. AB - Regulation of zinc metabolism by dibutyryl cAMP, glucagon, and epinephrine was examined in rats fed adequate amounts of zinc. Dibutyryl cAMP, epinephrine, and glucagon each produced an increase in liver metallothionein levels by 10 h after they were first administered. The increase in liver metallothionein was inversely related to the serum zinc concentration. Treatment with dexamethasone, a glucocorticoid, accentuated these effects to some extent. Both metallothionein I and II were induced by dibutyryl cAMP and glucagon. Levels of metallothionein mRNA in total liver RNA extracts were measured by dot blot hybridization using a synthetic 21-base oligonucleotide complimentary to the 5' region of both the metallothionein I and II genes. Individual administration of dibutyryl cAMP, glucagon, and epinephrine increased the number of metallothionein mRNA molecules per cell by up to fourfold. The data suggest that glucagon and epinephrine are primary regulators of metallothionein gene expression acting at least in part via cAMP. In adrenalectomized rats, glucagon, dibutyryl cAMP, and epinephrine had a less potent effect in terms of metallothionein induction and depression of serum zinc concentrations. These effects were largely restored when dexamethasone was also given. Collectively these data suggest that changes in zinc metabolism associated with acute stress involve coordinate regulation mediated by many factors, including glucocorticoids and cAMP. PMID- 3024500 TI - Insulin-sensitive glucoreceptors in rat preoptic area that regulate FFA mobilization. AB - Previous studies indicated that a longitudinal pathway connecting preoptic, lateral, and posterior hypothalamic areas participates in the process of free fatty acid (FFA) mobilization in the rat. In the present experiments, the presence of sensitive neurons in the preoptic area was investigated by examining the effects of topical stimulation with 2-deoxyglucose (2-DG) or insulin on the levels of plasma FFA in conscious unrestrained rats. Microinjections of minute amounts (50 micrograms and 1 microliter) of 2-DG into the preoptic area of fed animals induced rapid increases in the concentration of plasma FFA. Microinjections of insulin (5 microU and 0.5 microliter) produced sharp decreases of the elevated plasma FFA levels in fasted rats. Both 2-DG and insulin induced small increases in plasma glucose that did not differ from similar increases induced by equal volumes of 0.15 M NaCl. The results provide direct evidence for the presence within the preoptic area of insulin-sensitive glucoreceptors involved in FFA mobilization. The data suggest that activation of these receptors and increased sympathetic outflow to adipose tissue contributes to fasting lipolysis. PMID- 3024501 TI - Lower esophageal sphincter relaxation is associated with increased cyclic nucleotide content. AB - Experiments were conducted to determine whether relaxation of the opossum isolated lower esophageal sphincter (LES), induced by electrical field stimulation (EFS) or various pharmacological agents, is associated with changes in cyclic nucleotide content. EFS relaxed the LES in a frequency-dependent manner with 0.7 Hz producing half-maximal relaxation. Control tissues and tissues stimulated at various frequencies were clamp-frozen and assayed for cyclic nucleotide content. EFS had no effect on adenosine 3',5'-cyclic monophosphate (cAMP) content but increased guanosine 3',5'-cyclic monophosphate (cGMP) content in a frequency-dependent manner. Tetrodotoxin eliminated both the relaxation and cGMP accumulation in response to EFS. Vasoactive intestinal polypeptide (VIP) relaxed the LES with an EC50 of 0.1 microM. In contrast to the results with EFS, VIP enhanced cAMP content but had no effect on cGMP content. Relaxation of the LES produced by sodium nitroprusside or atriopeptin II was accompanied by an increase in cGMP accumulation, whereas isoproterenol- and dopamine-induced relaxation was accompanied by an increase in cAMP content. The data indicate that, depending on the stimulus, increases in either cAMP or cGMP content can accompany LES relaxation. These results are consistent with the proposed role of cyclic nucleotides as second messengers mediating LES relaxation. PMID- 3024502 TI - Cyclic nucleotide-dependent protein kinases in the lower esophageal sphincter. AB - The characteristics and regulation of adenosine 3',5'-cyclic monophosphate (cAMP) dependent and guanosine 3',5'-cyclic monophosphate (cGMP)-dependent protein kinases (PKs) in opossum, canine, and human lower esophageal sphincter (LES) were investigated. As measured by the incorporation of 32P from [gamma-32P]adenosine 5'-triphosphate (ATP) into histone, LES homogenates from all three species contained three distinct types of PK: cAMP-dependent PK, cGMP-dependent PK, and cyclic nucleotide-independent PK. In all three species, cAMP-dependent PK comprised approximately 80%, cGMP-dependent PK comprised approximately 10%, and cyclic nucleotide-independent PK comprised approximately 10% of the total PK activity in the LES. Diethylaminoethyl-sepharose column chromatography of the supernatant fraction of opossum LES homogenates revealed that of the total cAMP dependent PK, 10% was Type I and 90% was Type II. In contrast, equal amounts of Type I and Type II cAMP-dependent PK were present in both the human and canine LES. Isoproterenol-induced relaxation of the isolated opossum LES was accompanied by an increase in cAMP content and an activation of cAMP-dependent PK. The results of this study support the proposal that cyclic nucleotides and cyclic nucleotide-dependent PKs regulate LES tone. PMID- 3024503 TI - A sodium-hydrogen exchange system in isolated apical membrane from LLC-PK1 epithelia. AB - We have monitored transmembrane pH gradients using acridine orange fluorescence quenching and traced Na+ flux to study the properties of Na+-H+ exchange in apical membrane vesicles isolated from LLC-PK1 epithelia. The membranes have low conductance for Na+, H+, and K+ ions. An outwardly directed K+ gradient in the presence of valinomycin and carbonyl cyanide p-trifluoromethoxyphenyl hydrazone produced intravesicular acidification. This pH gradient was collapsed by addition of extravesicular Na+ or Li+ ions but not by tetramethylammonium. Amiloride (10( 4) M) inhibited the effect of both Na+ and Li+. An outwardly directed Na+ gradient stimulated H+ influx, which was also inhibited by 10(-4) M amiloride. Membrane short-circuit conditions affected neither Na+ nor H+ flux, consistent with transport mediated by an electroneutral process. The interaction of amiloride and sodium is consistent with noncompetitive inhibition with Ki = 100 +/- 10 microM for amiloride and an apparent Km for Na+ of approximately 20 mM. This finding is in agreement with previous studies of intact LLC-PK1 epithelia but differs from observations in brush-border membrane vesicles isolated from kidney proximal tubule in which competitive and mixed inhibition have been reported. These observed differences can be reconciled if two types of Na+-H+ exchange systems exist along the nephron, one with competitive and the other with noncompetitive inhibition, and if only the latter is expressed in the homogeneous cultured cells. PMID- 3024504 TI - Vasopressin-induced increases of cytosolic calcium in LLC-PK1 cells. AB - LLC-PK1 cells, a permanent cell line of renal tubule origin, which has vasopressin (VP) receptors and an adenosine 3',5'-cyclic monophosphate (cAMP) response to VP were grown to confluence on glass cover slips and loaded for 30-45 min with fura-2. Exposure to fura-2 did not affect cell viability (greater than 99%), K+, or ATP levels. Free cytosolic Ca2+ (Caf) was estimated spectrofluorometrically on washed cover slips. Basal levels averaged 73 +/- 3 nM. Peak Caf levels induced were: 10(-8) M VP - 128 +/- 24 nM, 10(-7) M VP - 301 +/- 51 nM, 10(-6) M VP - 537 +/- 117 nM. Peak Caf after 10(-6) M VP was reached in 42 +/- 5 s, followed by a return toward basal levels. Addition of a second dose of 10(-6) M VP following an initial dose of 10(-6) M VP did not raise Caf. Chelation of medium Ca2+ with ethyleneglycol-bis(beta-aminoethylether)-N,N'-tetraacetic acid just prior to 10(-6) M VP did not reduce the response of Caf (peak of 359 +/ 53). In contrast to VP, 10(-6) M calcitonin and PTH did not significantly increase Caf. The response to 10(-6) M VP was not significantly modified by prior prostaglandin E2 (3 microM) or dibutyryl-cAMP (100 microM). The response to 1 desamino-8-D-arginine vasopressin was less than that to VP. However, studies with VP-receptor antagonists did not allow definitive delineation of receptor type. These data provide evidence for VP-induced increases of Caf via release from intracellular stores in a renal epithelial cell. PMID- 3024505 TI - Enhanced cAMP response to vasopressin in the CCT of DOCA-Na hypertensive rats. AB - We characterized altered adenosine 3',5'-cyclic monophosphate (cAMP) regulation in deoxycorticosterone acetate (DOCA)-Na hypertensive rats using endogenous cAMP accumulation in the intact cell system of microdissected renal tubule fragments. Increased cAMP accumulation in response to vasopressin (VP) in cortical collecting tubules (CCT) began on day 5 (67%) after exposure to DOCA-Na and increased by 320% on day 42. Increased blood pressure began after day 7 and polyuria after day 17. The increased response to VP was DOCA dependent and appears to be exaggerated by dietary NaCl. Anatomic and hormone specificity studies were done on days 21-30. These included cAMP responses to prostaglandin E2, parathyroid hormone, thyrocalcitonin, VP, and isoproterenol in the CCT. The cAMP response to VP was measured in the glomerulus, proximal convoluted tubule, thin descending limb of Henle, medullary thick ascending limb of Henle, cortical thick ascending limb of Henle, medullary collecting tubule, and CCT. The supersensitivity occurred only to VP and only in the CCT. Thus this alteration in the VP response is anatomic and hormone specific and does not appear to be an acute effect of DOCA, since it was not present on day 1 and on day 3 of DOCA exposure. DOCA-Na hypertension is VP dependent. A specific exaggerated cAMP response to this hormone in the CCT would be expected to cause increased sodium retention. Whether increased sodium retention at this site contributes to hypertension in the DOCA-Na rat is unknown. PMID- 3024506 TI - Electrical properties of sodium bicarbonate symport in kidney epithelial cells (BSC-1). AB - Intracellular potentials of BSC-1 kidney epithelial cells known to express a Na+ HCO3- symport ranged between -40 and -70 mV, mean value Vm = -55.1 +/- 10.1 mV. Lowering HCO3- at constant partial pressure of CO2 (PCO2) or complete removal of HCO3-/CO2, rapidly depolarized, whereas readding HCO3- hyperpolarized Vm (40 +/- 8 mV/decade HCO3- at constant PCO2). This response was largely reduced by 1 mM 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS) (which depolarized Vm to about -20 mV) and in Na+-free medium, but Ba2+ had no effect. In HCO3(-)-Ringer, Na+ removal rapidly depolarized Vm (by 32 +/- 7 mV), and readdition hyperpolarized Vm. This was reduced in HCO3(-)-free medium or by 1 mM DIDS, but not by amiloride (10(-5) and 10(-3) M) or ouabain (10(-4) M). In the absence of HCO3- and/or Na+, steady-state Vm was reduced to -12 +/- 5 mV. Cl- removal had no effect on the responses to Na+ and/or HCO3- and led to a slow steady-state depolarization. Both in the presence and absence of HCO3-, raising pHi (changed by NH4Cl or butyrate) depolarized, whereas lowering pHi hyperpolarized Vm. Lowering pHo in HCO3(-)-free Ringer depolarized Vm (23 +/- 4 mV/decade H+). The slope conductance for K+ is only 6 +/- 2 mV/decade. Thus BSC-1 cells display typical electrical characteristics of Na+-HCO3- symport. In contrast to other systems, the data are compatible with a net electrogenic inward transport of Na+ and HCO3-. There might be an additional H+-(OH-) conductance operating also under nominally bicarbonate-free conditions. PMID- 3024507 TI - Calmodulin: biochemical, physiological, and morphological effects on Schistosoma mansoni. AB - Results of radioimmunoassays for the Ca2+-binding protein, calmodulin, revealed that this receptor constitutes 0.53 +/- 0.12% of the total protein in adult male Schistosoma mansoni. Schistosome calmodulin purified by Ca2+-dependent hydrophobic interaction chromatography showed an apparent molecular weight of 19 kDa, and its mobility on sodium dodecyl sulfate polyacrylamide gels was influenced by the presence of Ca2+ but not the antischistosomal drug praziquantel. Calmodulin from the parasite effected a four-fold stimulation of bovine heart adenosine 3',5'-cyclic monophosphate phosphodiesterase; this process was inhibited by removal of Ca2+ with ethyleneglycol-bis(B-aminoethylether)-N,N' tetraacetic acid but not by praziquantel. Inhibition of calmodulin-activated processes with antipsychotic compounds in vitro resulted in a number of time- and concentration-dependent changes, including inhibition of schistosome calmodulin stimulation of bovine heart phosphodiesterase, disruption and depolarization of the parasite's tegument, and positive inotropic effects on longitudinal musculature. Results of this study indicate that calmodulin is a functional component of schistosomes and suggest that the role it serves is analogous to that obtained in other eukaryotes; i.e., it is an important component of numerous processes regulated, in part, by Ca2+. PMID- 3024508 TI - Insulin and metabolic efficiency in rats. I. Effects of sucrose feeding and BAT axotomy. AB - The role of insulin and brown adipose tissue (BAT) thermogenesis in metabolic efficiency (ME, the efficiency of body wt gain) was examined in rats with varied basal insulin status. Long-lasting insulin was administered using a protocol that did not alter food intake, yet increased ME in both groups. Half the rats were fed sucrose to stimulate BAT growth and thermogenesis. Insulin overrode the exaggerated decrease in ME in sucrose-fed diabetics, with only partial attenuation in controls. Interscapular BAT (IBAT) lipoprotein lipase activity was decreased in diabetic rats, restored by insulin treatment, and not affected in controls. Sucrose-fed diabetics and controls had their IBAT sham or bilaterally surgically denervated. Insulin decreased the thermogenic potential of BAT [cytochrome oxidase activity (COA)] in intact controls and diabetics; in the latter, insulin restored COA independent of BAT innervation. We conclude that insulin can increase ME without an associated increase in energy intake, regardless of basal insulin status, both insulin deficiency and excess decrease BAT thermogenic potential (COA), and hyperinsulinemia-induced increases in ME may result from decreased BAT mitochondrial proliferation. PMID- 3024509 TI - Insulin and metabolic efficiency in rats. II. Effects of NE and cold exposure. AB - The role of insulin in metabolic efficiency (ME, i.e., efficiency of body wt gain) was examined under conditions of maximal energy expenditure in control and diabetic rats. Long-lasting insulin was administered using a protocol that did not affect food intake and increased ME in both groups. Half the animals were injected chronically with norepinephrine (NE). NE alone in controls decreased body weight and ME and increased brown adipose tissue (BAT) growth, thermogenic potential [cytochrome c oxidase activity (COA)], and lipoprotein lipases (LPL) activity; however, in diabetics, body weight, ME, and food intake all decreased and only BAT LPL activity and DNA content increased. The combination of NE and insulin increased BAT protein and COA in diabetics; in controls, all BAT measures were further increased and ME was intermediate to that of either treatment alone. Cold exposure decreased body weight and ME, increased food intake and qualitatively produced similar increases in BAT growth, COA, and LPL activity in both controls and diabetics. In diabetics, combined cold exposure and insulin did not affect the increase in BAT growth or LPL activity resulting from either treatment alone, but in controls this combination decreased BAT growth and COA. It is concluded that, even under conditions of maximal energy expenditure, both extremes of basal insulin status result in decreased BAT growth and thermogenic potential, but have opposite effects on ME. PMID- 3024510 TI - Liver glycogenolysis during exercise in adrenodemedullated male and female rats. AB - Previous studies have shown that adrenodemedullation has no effect on the rate of liver glycogenolysis during exercise in male rats. Mature female rats have been reported to have a higher hepatic beta-adrenergic receptor activity than do male rats of the same age. The present study was undertaken to determine the role of plasma epinephrine in stimulating liver glycogenolysis during exercise in female rats. Both male and female rats were adrenodemedullated or sham operated. Three weeks later rats were run for 60 min at 21 m/min up a 15% grade. The rate of liver glycogenolysis during exercise was not affected by adrenodemedullation in either female rats or male rats. Hepatic adenosine 3',5'-cyclic monophosphate increased to approximately the same extent in sham operated as in adrenodemedullated female rats during exercise. Adrenodemedullation caused a significant reduction in the amount of glycogen utilized by the soleus muscle and in the degree of hyperglycemia during exercise. We conclude that epinephrine is unessential for stimulation of liver glycogenolysis during exercise in either male or female rats. PMID- 3024511 TI - Possible mechanism for increased beta-adrenergic dipsogenic response in food deprived rats. AB - The dipsogenic responsiveness to isoproterenol was studied in food-deprived male rats. Unstimulated water intake was similar between control and fasted groups, and parallel dose-response curves for the dipsogenic response induced by isoproterenol (10-50 micrograms/kg) were observed. A twofold elevation in dipsogenic responsiveness was observed in the fasted rats, and this enhanced response was correlated with a dose-dependent increase in plasma renin activity when compared with the control rats after administration of isoproterenol. beta Adrenergic receptor binding assays were performed on both heart and renal cortical tissues. In the heart the receptor concentration was decreased after food deprivation, whereas the affinity of the receptor for the beta-adrenergic antagonist [125I]CYP remained unchanged. On the other hand, an increased beta adrenergic receptor concentration without change in affinity was found in renal cortices of fasted rats. Results from these receptor binding studies correlated with the attenuated chronotrophic response and the increased dipsogenic response to beta-adrenergic stimulation in the fasted rat. Therefore stimulation of these increased renal receptors by isoproterenol could result in an enhanced activation of the renin-angiotensin system and thus be a factor responsible for the increased dipsogenic response induced by isoproterenol observed in the fasted rats. PMID- 3024512 TI - Pituitary-adrenal responses to repeated small hemorrhage in conscious dogs. AB - Potentiated pituitary-adrenocortical responses to the second of two identical small hemorrhages, spaced 24 h apart, are seen in the pentobarbital sodium anesthetized dog. To investigate the role of pentobarbital anesthesia in these results and to better define the range of the effect, we studied awake trained dogs with chronic adrenal venous catheters. Each dog was bled an amount between 8.7 and 21.8% of measured blood volume [131I] (MBV) over 3 min, and peripheral and adrenal blood were sampled. Blood was reinfused 1 h later, and the dogs were fed. The same hemorrhage and experimental protocol were repeated 24 h later. Steroids were assayed by high-performance liquid chromatography-ultraviolet (HPLC UV) and adrenocorticotropic hormone (ACTH) by radioimmunoassay (RIA). Secretory rates of cortisol were calculated using measured adrenal blood flow rates. Maximal secretion of cortisol was determined after injection of 100 mU ACTH following each experiment. Dogs whose day 1 cortisol secretion after hemorrhage was submaximal (hem volume = 14.8 +/- 3.7% MBV; n = 7) showed a greater cortisol secretory response to the same hemorrhage on day 2 (P less than 0.005). This increased cortisol response on day 2 was accompanied by an increased ACTH presentation rate (APR) (P less than 0.025) and by increased adrenal sensitivity to ACTH (P less than 0.025). The increased APR was caused by both an increased venous ACTH and by an increased adrenal blood flow. If posthemorrhage cortisol secretion was maximal on day 1, ACTH, APR, and ABF were not different on the 2 days. No hemodynamic differences were seen to explain these findings. These results confirm and extend our previous results.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3024513 TI - Vasopressin responses to corticotropin releasing factor and hyperosmolality in conscious dogs. AB - We recently reported that ovine corticotropin releasing factor (CRF) infusion in conscious dogs elevated plasma vasopressin. The present study examines the vasopressin, adrenocorticotropic hormone (ACTH), and cortisol responses to CRF infusion (20 ng X kg-1 X min-1), to hypertonic saline infusion (NaCl 0.054 meq X kg-1 X min-1), and to simultaneous coinfusion of CRF and NaCl (CRF + NaCl) without (no-dex) or with (dex-treated) dexamethasone pretreatment in six conscious dogs (6-8 experiments/dog). CRF had no significant effect on plasma sodium or osmolality, blood pressure, or heart rate. NaCl increased plasma sodium from 146 +/- 1 to 151 +/- 1 meq/l and plasma osmolality from 298 +/- 3 to 305 +/- 3 mosmol/kg. Vasopressin increased significantly during CRF (2.1 +/- 0.5 to 4.8 +/- 1.1 pg/ml) and NaCl (1.9 +/- 0.3 to 5.0 +/- 0.8 pg/ml). Coinfusion of CRF and NaCl resulted in a response larger than the sum of the two infusions alone (3.0 +/- 1.6 to 31.4 +/- 18.5 pg/ml). The ACTH response to CRF (45 +/- 8 to 288 +/- 88 pg/ml) was not augmented by coinfusion with NaCl. DEX attenuated the vasopressin and ACTH responses to each infusion. We conclude that CRF-induced increases in vasopressin are augmented by a simultaneous osmotic stimulus. In addition, the plasma vasopressin responses to CRF and/or hypertonic saline infusion are inhibited by glucocorticoid pretreatment. PMID- 3024514 TI - Serum antibodies to Epstein-Barr virus in patients with major depressive disorder. AB - To determine whether major depressive disorder might be associated with serologic evidence for a chronic active Epstein-Barr virus infection, viral-specific antibodies were measured in two separate groups of depressed patients (N=43) and in 46 appropriately matched healthy volunteers. No evidence that depression affects cellular immunity to the point that a persistent Epstein-Barr virus carrier state becomes activated was found. There was also no evidence that depression results from an unrecognized chronic active Epstein-Barr virus infection. The authors conclude that the routine clinical determination of expensive commercial Epstein-Barr virus antibody profiles is not indicated in most patients with major depressive disorder in the absence of other signs of chronic active Epstein-Barr viral infection. PMID- 3024515 TI - Hereditary gastrointestinal polyposis syndromes. AB - Hereditary gastrointestinal polyposis syndromes can be divided into adenomatous and hamartomatous types. Familial adenomatous polyposis coli (FAPC) is the prototype adenomatous polyposis syndrome and is defined by the autosomal dominant transmission of multiple (more than 100) colorectal adenomas. Virtually all affected patients develop colorectal carcinoma if untreated. Adenomas may develop also in the stomach and small bowel in FAPC patients, but the incidence of carcinoma in these sites is low. A variety of extracolonic manifestations has been reported in FAPC, with the name Gardner's syndrome applied to kindreds with osteomas of the skull and mandible, multiple epidermal cysts, and other skin and soft-tissue lesions. In Turcot's syndrome, brain tumors are present. The distinction between Gardner's and Turcot's syndromes and classical FAPC has become blurred because of marked overlap between them; some authorities consider them to be varying manifestations of a single genetic defect. The hamartomatous polyposes include Peutz-Jeghers syndrome, familial juvenile polyposis, Cowden's disease, intestinal ganglioneuromatosis, and the Ruvalcaba-Myrhe-Smith syndrome. The incidence of gastrointestinal cancer in patients with Peutz-Jeghers syndrome and familial juvenile polyposis exceeds that in the normal population, but is relatively low. In Cowden's disease, the gastrointestinal tract may be the site of multiple hamartomas, but there is no associated increase in the incidence of gastrointestinal cancers; instead, there is an increased incidence of carcinoma of the breast and thyroid. Intestinal ganglioneuromatosis occurs in von Recklinghausen's disease, in association with multiple endocrine neoplasia, type 2b, or as an isolated abnormality. Patients with ganglioneuromatosis do not appear to have an increased risk of developing gastrointestinal cancer. Ruvalcaba Myrhe-Smith syndrome comprises macrocephaly, mental deficiency, an unusual craniofacial appearance, hamartomatous intestinal polyposis, and pigmented macules on the penis. No increased risk of developing cancer has been identified in the few reported cases. PMID- 3024517 TI - [Changes in the activity of natural killer cells in patients with benign and malignant ovarian neoplasms]. PMID- 3024516 TI - Biology of Arboledas virus, a new phlebotomus fever serogroup virus (Bunyaviridae: Phlebovirus) isolated from sand flies in Colombia. AB - Six isolates of a new phlebotomus fever serogroup virus, designated Arboledas virus, were obtained from sand flies (Lutzomyia spp.) collected in northeastern Colombia. One of the isolates was made from a pool of male sand flies. By immunofluorescence, Arboledas virus is related to Caimito and Pacui viruses; by neutralization test, it is distinct. Arboledas virus neutralizing antibodies were found in the sera of opossums (Didelphis marsupialis) and humans living in the study area. D. marsupialis inoculated with the virus developed a viremia of four days' duration, and sand flies (Lutzomyia gomezi) feeding on a viremic opossum were readily infected. Transovarial transmission of Arboledas virus was also demonstrated in experimentally infected Lu. gomezi. Results of the above laboratory studies suggest that Arboledas virus is maintained in nature by two mechanisms: vertical (transovarial) transmission in the insect vector, and an alternating marsupial-sand fly cycle. The implications of this complex maintenance cycle for other phleboviruses are discussed. PMID- 3024518 TI - [Use of the muscle relaxant arduan in obstetric and gynecologic operations]. PMID- 3024519 TI - Effects of nedocromil sodium and placebo delivered by pressurised aerosol in bronchial antigen challenge. PMID- 3024520 TI - Exercise-induced anaphylaxis: inhibition with sodium bicarbonate. PMID- 3024521 TI - Effect of AA-861, a 5-lipoxygenase inhibitor, on leukotriene synthesis in human polymorphonuclear leukocytes and on cyclooxygenase and 12-lipoxygenase activities in human platelets. AB - AA-861, a selective inhibitor of 5-lipoxygenase of arachidonic acid, was tested for ability to inhibit leukotriene C4 and leukotriene B4 synthesis in human polymorphonuclear leukocytes after calcium ionophore stimulation. AA-861 dose dependently inhibited leukotriene B4 and leukotriene C4 generation in human polymorphonuclear leukocytes; the concentration required to inhibit generation by 50% (IC50) was 3 X 10(-7) M for leukotriene B4 and 1 X 10(-8) M for leukotriene C4. BW-755C inhibited the generation of leukotriene C4 with an IC50 of about 10( 5) M, indicating that AA-861 is about 1,000 times more potent than BW-755C. AA 861 did not affect the activity of either cyclooxygenase or 12-lipoxygenase at a concentration up to 10(-5) M in human platelets. AA-861 did not inhibit histamine release from human basophils. These results indicate that AA-861 selectively inhibits 5-lipoxygenase but not cyclooxygenase or 12-lipoxygenase in human specimens. PMID- 3024522 TI - [Glomangioma or glomus tumor of the nasal vestibulum]. PMID- 3024523 TI - The management of labour using continuous lumbar epidural analgesia in a patient with a malignant cerebral tumour. AB - A mother presented at 28 weeks with epileptiform convulsions which were due to a malignant cerebral tumour. Labour and delivery at 34 weeks were managed under epidural analgesia. The infant was healthy, but the mother died several days later following cerebral decompression. PMID- 3024524 TI - Derivatization of ethylene dibromide with silica-supported silver picrate for improved high-performance liquid chromatographic detection. PMID- 3024525 TI - Alkaline phosphatase and 5'-nucleotidase enzymes in the carotid rete-cavernous sinus complex of sheep and goats. AB - 12 sheep and 4 goats were used to detect the presence of alkaline phosphatase (A.P.) and 5'-nucleotidase (5'-N) enzymes in the carotid rete-cavernous sinus structure. Different methods of preservation were used. The calcium and the lead methods were used to detect the presence of A.P. and 5'-N, respectively. Best results in their detection were obtained with liquid nitrogen preservation. A.P. enzyme was found in and around areas in which blood capillaries were present, indicating active transport of materials through the capillary membrane. Slight enzymatic activity was seen on the endothelial surface of the rete branches, while the enzyme seemed to be absent from the cavernous sinus. 5'-N was discernible in the tunica adventitia and in the endothelial cells, while the tunica media of the rete branches was apparently devoid of this enzyme. Possible role of these enzymes in the vascular wall metabolism of this structure has been discussed. PMID- 3024526 TI - Cytomegalovirus infections: occupational risk for health professionals. AB - Health care personnel are becoming increasingly aware of potential hazards associated with caring for patients with contagious diseases. The cytomegalovirus is of special concern, because infection with this virus in a pregnant female employee could be associated with significant neurologic injury in her fetus. Nosocomial transmission from patient to health care worker has not been documented. A review of cytomegalovirus excretion in hospitalized patients and prospective evaluations of primary infection in hospital personnel do not support frequent occurrence of cytomegalovirus infection despite ample opportunity for exposure. Adherence to proper isolation techniques should be adequate to prevent nosocomial transmission of the cytomegalovirus. PMID- 3024527 TI - Isocyanate-induced respiratory disease. PMID- 3024528 TI - [Distribution of different phenotypes of paraoxonase in a French population]. AB - Earlier studies show that there is a polymorphism of the paraoxonase and that the heterozygous phenotype can be distinguished from both homozygous phenotypes, by studying the ratio of paraoxonase to arylesterase activities. The separation of the three phenotypes of the paraoxonase was possible on our population using the same techniques as Eckerson et al. and by a statistic method from Schroeder, 32 families have been studied. The gene frequencies for the A and B alleles are: 0.6875 and 0.3125 respectively in a french population. They are very close to those of an American Caucasian population: A: 0.685 and B: 0.315 of Eckerson et al. PMID- 3024529 TI - [Collagenases in rheumatology]. AB - An important role is supposed to be played by the collagenases in the physiological or pathological degradation of collagen. However, their exact effects in rheumatic diseases, especially in rheumatoid arthritis is still controversial. The aim of the authors is to give facts and datas up to date, about this important question of the pathogenic role of collagenases in rheumatoid arthritis, osteoarthritis and in several other rheumatic diseases. PMID- 3024530 TI - [Cartilage and collagenases]. AB - The articular cartilage is frequently destroyed in rheumatic diseases. Among the various responsible factors, enzymes and especially collagenases play a leading role. Their action explains the collagen degradation linked with cartilage destruction. The author recalls the origin, mechanism of action of the articular collagenases and discusses their role in the articular cartilage degradation in rheumatic diseases. PMID- 3024532 TI - An improved method for phenotyping individuals for the human serum paraoxonase arylesterase polymorphism. AB - Human serum paraoxonase/arylesterase is a polymorphic enzyme, determined by two allelic genes at one autosomal locus. These two isozymes are called (A) and (B), and the three corresponding phenotypes A, AB and B. We measure paraoxon hydrolysis (paraoxonase activity), with, or without 1 M NaCl, and phenylacetate hydrolysis (arylesterase activity) with, or without 0.1 mM chlorpromazine. The resulting four measurements make it possible to determined essentially every individual's phenotype. The combination of qualitative tests, based on distinctive characteristics of the two isozymes, greatly improves our ability to determine individual phenotypes and detect other isozymic variants of the esterase. For several years, we have been interested in the possible correlation of the phenotypes of paraoxonase/arylesterase and sensitivity or resistance to organophosphate exposure [1]. Recent reports show that the paraoxonase gene may be closely linked to the gene for cystic fibrosis [2]. If this will be confirmed, it could be another very practical reason for typing people for this polymorphic marker. Being able to identify those heterozygous for the paraoxonase/arylesterase polymorphism, about 43% of the European population, should greatly enhance the value of phenotyping for this simple, convenient, polymorphic trait. PMID- 3024531 TI - [Collagen and steroid hormones]. AB - The authors analyse the effects of steroid hormones on collagen, from up to date datas previously published and personal works. Glucocorticoids have catabolic effects; their molecular effects are reviewed. Conversely, oestrogens and androgens have an anabolic effect. PMID- 3024533 TI - [Simultaneous presence of isoenzymes CK-BB and CKm of serum creatine kinase in small-cell bronchopulmonary cancer. Limitations of the use of M subunit immuno inhibition technics]. PMID- 3024534 TI - Detection of latent pseudorabies virus in porcine tissue, using a DNA hybridization dot-blot assay. AB - A DNA-hybridization dot-blot technique was developed to detect the presence of pseudorabies virus (PRV) DNA in porcine tissue. Seven 32P-nick translated probes of high specific activity were prepared from transformed Escherichia coli plasmids into which Bacillus amyloliquefaciens H (Bam HI) restriction fragments of PRV-DNA had been inserted. Samples of DNA that had been extracted from porcine tissue or from PRV grown in tissue culture were transferred to nitrocellulose paper, using a microsample filtration manifold and were hybridized to the probes under high-stringency conditions. Under optimal hybridization conditions, the minimum detection amount of PRV-DNA was 10(-11) g, which is equivalent to 1 copy of the PRV genome/80 host cells. Four probes did not show cross hybridization with DNA extracted from tissues of known PRV-negative swine, and these were subsequently used to detect PRV-DNA in infected porcine tissues. Generally, correlation between virus isolation and hybridization data was good for tissues from swine that had died of acute PRV infection. Furthermore, PRV-DNA was present in specific tissues of all 4 seropositive swine that had recovered from pseudorabies and in which no infective virus or viral products were detected at necropsy. Pseudorabies virus DNA was present in the rostralis cerebral cortex (n = 2) or in the medulla oblongata (n = 1) and trigeminal ganglion (n = 1). This probably indicated the portal of entry of the virus into the CNS. In another seropositive pig, there was evidence of a productive infection in the tonsils, although virus was not isolated in a tissue culture system.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3024535 TI - In vitro induction of cytotoxic lymphocytes from infectious bovine rhinotracheitis virus hyperimmune cattle. AB - Cytotoxic lymphocytes against infectious bovine rhinotracheitis virus (IBRV) infected cells were induced by in vitro stimulation with IBRV antigen of peripheral blood leukocytes obtained from hyperimmune cattle. After one in vitro stimulation with IBRV, cytotoxicity was detected against IBRV-infected autologous target cells, but not against heterologous IBRV-infected or K562 target cells. After 4 in vitro IBRV stimulations at weekly intervals (in the presence of autologous feeder cells followed by interleukin 2 treatment at 2-day intervals), cytotoxicity was detected against autologous-, and to a lesser extent, heterologous-infected cells. Most cells in cytotoxic cultures were positive to a monoclonal antibody shown to react with bovine T lymphocytes. PMID- 3024536 TI - Detection of transmissible gastroenteritis coronavirus antigens by a sandwich enzyme-linked immunosorbent assay technique. AB - A new sandwich enzyme-linked immunosorbent assay, using monoclonal and polyclonal antibodies, was developed to detect transmissible gastroenteritis virus antigens from cell culture and from intestinal wash or feces obtained from experimentally infected pigs. This technique was shown to be suitable for the detection of virulent field strain unadapted to cell culture. Cross reactions had not been observed with other enteric pathogens, rotavirus, porcine epizootic diarrhea virus, and Escherichia coli. PMID- 3024537 TI - Asbestos exposure and retention as determinants of airway disease and asbestos alveolitis. AB - To evaluate the relationships of asbestos exposure, retention, airway response, and the asbestos alveolitis, we exposed 2 groups of sheep every 2 wk for 3 yr to either 100 ml phosphate-buffered saline (PBS) or 100 mg UICC chrysotile fibers in 100 ml PBS. The sheep were evaluated periodically by pulmonary function tests (PFT), chest radiograph (CR), bronchoalveolar lavage (BAL), and transbronchial lung biopsy (TLB). At Month 24 of the study, all asbestos-exposed sheep had significant increases in lung resistance and upstream resistance. However, only 9 of the 16 asbestos-exposed sheep had significant changes in TLB, CR, Cst, and VC, which clearly separated them from the other 6 sheep in these parameters. The 2 groups, however, had similar air-flow limitation. At lung biopsy, all asbestos exposed sheep had significant peribronchiolar fibrosis, with significant alveolitis only in the group of 9 sheep with radiographic and functional changes of early asbestosis. The 9 sheep also had significant changes in BAL cellularity and biochemical profile, which differentiated them from the other 6 asbestos exposed sheep. Analysis of BAL fiber content at that point revealed that despite identical exposure, the group with interstitial lung disease had significantly more fiber retention (p less than 0.01). The data demonstrate that whereas asbestos airway disease appears to be primarily an exposure-dose-related response, the lung response appears to be more closely related to alveolar retention of the dust. PMID- 3024538 TI - Hydroxyurea inhibits airway hyperresponsiveness in guinea pigs by a granulocyte independent mechanism. AB - Exposure of guinea pigs to toluene diisocyanate (TDI) causes an increase in airway responsiveness to inhaled acetylcholine. This increased airway responsiveness is temporally associated with an increase in polymorphonuclear leukocytes (PMN) in the tracheal wall. To determine whether PMN play a mechanistic role in this increase in acetylcholine responsiveness, we studied the effects of PMN depletion on this response with 2 different cytotoxic drugs, hydroxyurea and cyclophosphamide. Airway responsiveness was measured in untreated, hydroxyurea-treated, or cyclophosphamide-treated animals while they breathed spontaneously or during mechanical ventilation through a tracheostomy. In untreated animals, exposure to TDI caused a significant increase in airway responsiveness to acetylcholine for both spontaneously breathing and anesthetized and ventilated animals. This TDI-induced increase in airway responsiveness was associated with a significant influx of PMN into both the extravascular and intravascular trachea. Treatment with hydroxyurea, to reduce PMN counts in the bloodstream to less than 200/mm3, inhibited both the TDI-induced increase in airway responsiveness and the TDI-induced influx of PMN into the trachea of both spontaneously breathing and mechanically ventilated animals. In mechanically ventilated animals, treatment with cyclophosphamide, until PMN counts in the bloodstream were less than 200/mm3, also inhibited the influx of PMN into the trachea but did not inhibit the TDI-induced increase in airway responsiveness. These results suggest that PMN are not necessary for the TDI-induced increase in airway responsiveness and that hydroxyurea inhibits this effect by a mechanism other than PMN depletion. PMID- 3024539 TI - Uptake of low density lipoproteins by the hamster lung. Interactions with capillary endothelium. AB - The mechanism by which the circulating low density lipoproteins (LDL) contribute to the lung surfactant cholesterol was investigated by perfusing the hamster lung in situ with LDL either radiolabeled or coupled to gold, or both. Part of [125I] LDL and [3H]-cholesterol LDL were taken up by a specific process which was time- and concentration-dependent and reached saturation within 20 to 30 min of perfusion. Competition experiments and removal of receptor-bound LDL by heparin suggested that about 50% of LDL uptake is receptor-independent. Experiments using double labeled LDL showed a preferential uptake of 3H-cholesterol versus 125I by the lung both in situ and in vivo. LDL-gold particles (LDL-Au), recirculated through the isolated lung, bound to the endothelial luminal plasma membrane and to features potentially involved in receptor-mediated endocytosis (coated pits, coated vesicles, lysosomelike structures) and in transcytosis (plasmalemmal vesicles). The results suggest that LDL uptake by the lung takes place by both receptor-mediated and receptor-independent mechanisms. Cholesterol may be in part transferred to the lung without the apoprotein moiety; the alveolar capillary endothelium appears to be the first monitor of this complex process. PMID- 3024540 TI - [Partially differentiated cystic nephroblastoma]. AB - Two new cases of cystic partially differentiated nephroblastoma, which were treated by nephrectomy, are reported. Their gross appearance is undistinguishable from a multilocular cyst, with the only difference being the presence of buds of blastema within the cystic septa. While cystic partially differentiated nephroblastoma as well as multilocular cyst have never been reported to be associated with local or metastatic recurrence and therefore their treatment consists of simple nephrectomy, the presence of blastema suggests a differentiates stage of Wilms' tumor, thus demanding a close surveillance of the patient. PMID- 3024541 TI - The metabolism of plutonium and related elements. International Commission on Radiological Protection. A report of a Task Group of Committee 2. PMID- 3024542 TI - Intravenous immune globulin for prevention of cytomegalovirus infection and interstitial pneumonia after bone marrow transplantation. AB - The effects of high doses of polyvalent intravenous immune globulin given for prophylaxis of cytomegalovirus infection and interstitial pneumonia in recipients of allogeneic marrow transplants were evaluated in a randomized controlled trial. Both symptomatic cytomegalovirus infection (21% compared with 46%, p = 0.03) and interstitial pneumonia (18% compared with 46%, p = 0.02) occurred less frequently in the recipients of intravenous immune globulin than in control patients. Prophylactic intravenous immune globulin was also associated with a lower incidence of graft-versus-host disease (34% in recipients compared with 65% in controls, p = 0.01), but its reduction in rates of interstitial pneumonia was independent of graft-versus-host disease and occurred in both patients with and without graft-versus-host disease. The high doses of immune globulin were well tolerated. Prophylactic intravenous immune globulin can modify the severity of cytomegalovirus infection and prevent interstitial pneumonia and possibly graft versus-host disease in patients having allogeneic marrow transplantation. PMID- 3024543 TI - Itraconazole for cryptococcal infection in the acquired immunodeficiency syndrome. PMID- 3024544 TI - Cytomegalovirus and acute cholecystitis. PMID- 3024545 TI - Central thrombocytopenia and liver disease. PMID- 3024546 TI - Bone marrow examination in small cell lung cancer. PMID- 3024547 TI - Immunization of children infected with human T-lymphotropic virus type III/lymphadenopathy-associated virus. Recommendations of the Immunization Practices Advisory Committee. Centers for Disease Control, Department of Health and Human Services. AB - This document is intended to summarize available information and to assist health care providers in developing policies for the immunization of children infected with human T-lymphotropic virus type III/lymphadenopathy-associated virus (HTLV III/LAV) [now known as the human immunodeficiency virus], the virus that causes acquired immunodeficiency syndrome (AIDS). These policies may vary depending upon the prevalence of HTLV-III/LAV infection and the incidence of vaccine-preventable diseases in the community, individual assessment of a child's health status, and the risks and benefits of immunization in a particular situation. This discussion considers the risks and benefits of immunization for children residing in the United States based on the risks of vaccine-preventable diseases and the prevalence of HTLV-III/LAV infection and is intended for use by health-care providers in the United States. The recommendations may not pertain to other countries with different risks of vaccine-preventable diseases and prevalence of HTLV-III/LAV infection among children. Since these recommendations are based upon information and knowledge available at this time, periodic reassessment and revision will be required as more data concerning risk and benefits associated with immunization of HTLV-III/LAV-infected children become known and as the prevalences of specific vaccine-preventable diseases and HTLV-III infection change. PMID- 3024548 TI - Monoclonal autoantibodies that react with multiple organs. Basis for reactivity. AB - We have been able to make monoclonal MOR antibodies in several different ways. First, we have been able to prepare MOR antibodies from mice with autoimmune disease and from humans with autoimmune disease. Second, we have prepared MOR antibodies from normal mice without autoimmune disease and from healthy humans without evidence of autoimmune disease. Third, we have prepared MOR antibodies by hybridoma technology and by transformation of lymphocytes with EBV. We have shown that MOR antibodies are common and are part of the host's normal B-cell repertoire. Our studies raise the possibility that most monoclonal autoantibodies, when extensively screened, will to a greater or lesser degree be of the MOR type. PMID- 3024549 TI - Postinfectious autoimmunity: two distinct phases of coxsackievirus B3-induced myocarditis. PMID- 3024550 TI - Diagnosis and treatment of small cell carcinoma of the larynx: a critical review. AB - Small cell carcinoma of the larynx is an uncommon neuroendocrine tumor with particular pathologic, therapeutic, and prognostic connotations. The first case of this lesion was observed in Canada in 1972. Fourteen cases of small cell carcinoma of the larynx were observed in the Ear, Nose, and Throat Department of Padua University in a series of 3,284 primary and secondary laryngeal and hypopharyngeal malignant neoplasms. This number constitutes the largest collection from a single institution in the literature and brings the total recorded cases to 66. The tumor is thought to arise from the argyrophilic Kulchitsky cells normally found in laryngeal mucosa. The diagnosis is based on the light microscopic appearance of the neoplasm and can be confirmed by electron microscopy. The differential diagnosis must be made from carcinoid, atypical carcinoid, small cell squamous carcinoma, small cell ductal carcinoma, lymphoma, mycosis fungoides, and metastatic lung small cell cancer. Systemic chemotherapy with radiation therapy is the accepted manner of treatment today. The survival of the patients treated with these modalities may be significantly improved, and some patients may be cured. PMID- 3024552 TI - On the assessment of the in vitro biopotency and site(s) of action of drugs affecting adrenal steroidogenesis. AB - Dispersed guinea-pig adrenal cells have been employed in the in vitro estimation of the biological potency and sites of action of drugs acting against the adrenal. The effect of 12 drugs on cortisol secretion from cells stimulated with adrenocorticotrophin (ACTH, 50 ng/L, a 95% saturating dose) has been tested. All the drugs depressed cortisol output in a dose-related fashion. The concentration of drug which inhibited secretion by 50% was (mumol/L, mean +/- SEM): etomidate 0.1 +/- 0.002; epostane 0.44 +/- 0.02: 17-ketotrilostane 0.55 +/- 0.04: trilostane 1.3 +/- 0.1: metyrapone 3.5 +/- 0.6: cyproterone acetate 4.6 +/- 0.2: megestrol acetate 11 +/- 2: danazol 22 +/- 2: aminoglutethimide 41 +/- 5: stanozolol 50 +/- 4: thiopentone 160 +/- 18: propofol 170 +/- 18. The sites of the anti-steroidogenic effect of seven of these drugs have also been established using a method based upon the sequential stimulation by the exogenous precursor steroids of the various steps leading to the biosynthesis of cortisol by adrenal cells. Propofol acts between ACTH binding and pregnenolone production, trilostane, megestrol acetate and cyproterone acetate are 3 beta-hydroxysteroid dehydrogenase inhibitors whereas metyrapone, etomidate and thiopentone act at 11 beta-hydroxylase. PMID- 3024551 TI - Demonstration of human papillomavirus capsid antigen in carcinoma in situ of the larynx. AB - We have undertaken a retrospective study to investigate the role of human papillomavirus (HPV) in the development of laryngeal carcinoma in situ (CIS). Sixty paraffin-embedded tissue specimens, collected from 20 patients with a history of laryngeal CIS, were examined for the presence of HPV capsid antigen. All but four individuals were men, with an average age at diagnosis of 64 years. An immunoperoxidase technique showed that 20 specimens from 14 patients contained detectable HPV capsid antigen. An independent evaluation for histopathologic features characteristic of HPV infection identified viral changes in the 14 patients as well as an additional two. No correlation was found between clinical course, as determined by histologic severity of vocal cord lesions, and presence of HPV. These results suggest that HPV should be considered an etiologic agent in the development of laryngeal CIS. PMID- 3024553 TI - Determination of urinary 3-methoxy-4-hydroxymandelic and 3-methoxy-4 hydroxyphenylacetic acids by fused silica capillary gas chromatography. PMID- 3024554 TI - Long-term effect of bran ingestion on lipid metabolism in healthy man. AB - The long-term effect of the addition of 20 g wheat bran to the diet is studied over 7 weeks in 4 healthy subjects. No change is found in plasma cholesterol and triacylglycerol. In this experimental design, we find no modification in the plasma levels of the various lipoproteins, except for VLDL cholesterol, which remains significantly lower after 7 weeks of wheat bran ingestion. PMID- 3024555 TI - Locus ceruleus lesions and eosinophilic inclusions in MPTP-treated monkeys. AB - Systemic administration of the recently discovered neurotoxin 1-methyl-4-phenyl 1,2,3,6-tetrahydropyridine (MPTP) produces severe clinical parkinsonism and degeneration of the substantia nigra in humans and monkeys. In previous studies, no convincing structural damage to nerve cells outside the substantia nigra could be demonstrated in subhuman primates. Using a protracted MPTP regimen and older animals, we now report locus ceruleus lesions and eosinophilic inclusion bodies in squirrel monkeys. The inclusions were seen only in areas where Lewy bodies are found in human Parkinson's disease. No such abnormalities were seen in control animals. These findings suggest that similarities between the neuropathology of MPTP-induced parkinsonism in the monkey and human Parkinson's disease are greater than first thought and increase the usefulness of the MPTP monkey model for research in Parkinson's disease. PMID- 3024556 TI - X-linked neuropathy: gene localization with DNA probes. AB - We used probes for DNA polymorphisms on the X chromosome to study genetic linkage in four families with X-linked neuropathy. Despite clinical variability, all four families showed the same linkage pattern. We found evidence in each family of linkage to the marker DXYS1 on the proximal long arm of the X chromosome, as reported by others. We also found linkage to p58-1 (DXS14) on the proximal short arm. We found only loose linkage or nonlinkage to nine other markers located elsewhere on the chromosome. Our analysis places the gene defect for this disorder in the region of DXYS1 and p58-1, near the centromere of the X chromosome. PMID- 3024557 TI - Subacute sensory neuronopathy secondary to dorsal root ganglionitis in primary Sjogren's syndrome. AB - Sensory neuropathies, particularly trigeminal neuropathy, have been recognized as neurological complications of Sjogren's syndrome, but the pathogenesis has not been established. We describe a woman with primary Sjogren's syndrome who developed a progressive debilitating subacute sensory neuronopathy. Results of electrophysiological studies were consistent with involvement of the trigeminal and dorsal root ganglia. A thoracic dorsal root ganglion biopsy showed lymphocytic infiltration and degeneration of ganglion cells. We believe that this is the first description of biopsy-documented dorsal root ganglionitis in a subacute sensory neuronopathy associated with Sjogren's syndrome and that the finding suggests an immunopathogenic basis. PMID- 3024558 TI - Transfection of human skeletal muscle cells with SV40 large T antigen gene coupled to a metallothionein promoter. AB - We have undertaken to increase the proliferative capacity of cultured human skeletal myocytes by transfection with a plasmid construct that contains the immortalizing and transforming large T antigen gene of simian virus 40 (SV40) under the control of a zinc-sensitive metallothionein promoter. This construct was chosen to permit rapid growth of transformants in zinc-containing medium, which induces high levels of T antigen expression, and muscle-specific differentiation after withdrawal of exogenous zinc, which reduces levels of T antigen. When grown in 100 microM Zn2+, transformed myocytes expressed the large T antigen, divided rapidly, and acquired an apparently unlimited proliferative capacity. Transfer of these cells to a zinc-poor medium resulted in decreased T antigen immunofluorescence, growth rate, and saturation density as well as a return to a physiological spindle morphology. Despite transformation, these cells expressed differentiation markers characteristic of myoblasts: the B isoform of creatine kinase, and surface antigens 5.1H11, D5, and Thy 1 in the presence or absence of Zn2+. When grown to high density in a serum-poor medium, these cells differentiated further into typical multinucleated myotubes that expressed the M isoform of creatine kinase and increased levels of surface antigen 5.1H11, creatine kinase, and nicotinic acetylcholine receptors, but no detectable Thy 1 antigen. The specific activity of these differentiation markers was higher when the cells were grown in the absence of added zinc. These results indicate that transformation of human skeletal myocytes with a regulatable SV40 large T antigen gene allows an increase of the proliferative capacity of these cells with preservation of their capacity to differentiate in a physiological manner. PMID- 3024559 TI - [Inactivation characteristics of polyene macrolide antibiotics]. AB - Roseofungin, a pentaen was used as an example in investigation of certain characteristics of inactivation of polyenic antibiotics. An inactivation product of the antibiotic having no antiviral or antimycotic activity was isolated by TLC and column chromatography on Sefadex LH-20. UV, IR and EPR spectroscopy showed that this product was cys-tetraen without conjugation of the lactone carbonyl to the polyenic system. PMID- 3024560 TI - In vitro and in vivo activity of LY 146032, a new cyclic lipopeptide antibiotic. AB - The in vitro activity of LY 146032, a cyclic lipopeptide antibiotic belonging to the class of agents designated A21978C, was compared with those of vancomycin, cefpirome, cefotaxime, and clindamycin against selected gram-positive bacteria. The new drug inhibited all staphylococcal isolates, including methicillin resistant strains, at concentrations of less than or equal to 1.0 microgram/ml. The activity of LY 146032 was comparable to that of vancomycin against most streptococci, but the latter demonstrated greater potency against Streptococcus faecium and penicillin-resistant strains of pneumococci and viridans group streptococci. LY 146032 was markedly less active than vancomycin against Listeria monocytogenes (MICs for 90% of strains tested, 16 and 1.0 microgram/ml, respectively). The activity of LY 146032 was enhanced as the concentration of calcium in the test medium was increased. MBCs were within eightfold of the MIC for each of 12 strains tested. In a rat model of enterococcal endocarditis, the administration of LY 146032 resulted in increased survival and a reduction in the bacterial titer within cardiac vegetations compared with untreated control animals. PMID- 3024561 TI - Comparative methods for detection of thymidine kinase-deficient herpes simplex virus type 1 strains. AB - Four methods for analyzing viral susceptibility to antiviral substances were compared. In two methods viral products were measured: late viral proteins were measured by an enzyme-linked immunosorbent assay and viral DNA was measured by DNA hybridization. Infectious virus was quantified in the other two assays as the number of plaques and the yield of virus. The enzyme-linked immunosorbent assay procedure in our hands detected the smallest amounts (lowest proportions) of thymidine kinase-deficient herpes simplex virus type 1 mixed with wild-type virus. The thymidine kinase-deficient proportion of the herpes simplex virus type 1 isolate increased rapidly in the presence of acyclovir in cell culture. PMID- 3024562 TI - Mode of action, toxicity, pharmacokinetics, and efficacy of some new antiherpesvirus guanosine analogs related to buciclovir. AB - 9-[4-Hydroxy-3-(hydroxymethyl)butyl]guanine (3HM-HBG), (RS)-9-[4-hydroxy-2 (hydroxymethyl)butyl]guanine ([+/-]2HM-HBG), and cis-9-(4-hydroxy-2 butenyl)guanine (2EN-HBG), new acyclic guanosine analogs structurally related to buciclovir (BCV [(R)-9-(3,4-dihydroxybutyl)guanine]), were evaluated in parallel with buciclovir as anti-herpes simplex virus (HSV) agents. In cell cultures, replication of different strains of HSV type 1 (HSV-1) and HSV-2 was inhibited at nontoxic drug concentrations. The concentrations giving 50% inhibition of plaque formation were, however, dependent on virus strain and cell type. In most cell types, the order of activity against HSV-1 strains was 3HM-HBG greater than (+/ )2HM-HBG greater than BCV greater than 2EN-HBG, whereas the drugs showed an approximately equivalent activity against HSV-2 strains in different cells. The cytotoxic effects of the drugs were also cell type dependent, the order of activity being BCV greater than 3HM-HBG = (+/-)2HM-HBG greater than 2EN-HBG. At growth-inhibitory concentrations, the guanosine analogs BCV, 3HM-HBG, and (+/ )2HM-HBG showed clastogenic effects in human lymphocytes, mainly because of the induction of chromatid breaks. When evaluated for their anti-HSV effects in systemic HSV-1 infections in mice, the order of activity was BCV = 3HM-HBG greater than (+/-)2HM-HBG greater than 2EN-HBG, and in mice infected systemically with HSV-2, only BCV and 3HM-HBG showed efficacy. The differences between efficacy in vitro and in vivo could be explained in part by differences in kinetics of the drugs in mouse plasma, as the more efficacious drugs, BCV and 3HM HBG, showed lower clearances and longer half-lives than the less efficacious ones, (+/-)2HM-HBG and 2EN-HBG. When used topically against a cutaneous HSV-1 infection in guinea pigs, 3HM-HBG showed an effect equivalent to that of BCV, whereas (+/-)2HM-HBG and 2EN-HBG were inactive. Mechanistically, the guanosine analogs were characterized by a high affinity for the viral thymidine kinase and a low affinity fo a cellular thymidine kinase and by their inhibition of viral DNA synthesis in infected cells. PMID- 3024563 TI - Improved Listeria monocytogenes selective agar. AB - By increasing the LiCl concentration to 5 g/liter and adding 20 mg of moxalactam per liter to modified McBride agar base, it was possible to inhibit the growth of many bacteria which interfered with the recovery of Listeria monocytogenes from beef. PMID- 3024564 TI - Location of plasmid-mediated citrate utilization determinant in R27 and incidence in other H incompatibility group plasmids. AB - Citrate utilization (Cit+) is encoded by a specific subgroup of incompatibility HI plasmids, viz., IncHI1 plasmids. Only one IncHI1 plasmid, pRG1271, which originated in a Mexican typhoid outbreak in 1972, did not specify Cit+. All other Cit+ plasmids hybridized to a Cit+ probe, a 2-kilobase BglII fragment derived from the Cit+ transposon Tn3411. The position of the Cit+ determinant was mapped to a 13.5-kilobase ApaI fragment within the prototype IncHI1 plasmid R27. No other functions have been mapped within this region. Citrate utilization mediated by IncHI1 was observed only after a prolonged lag period of approximately 150 h, and certain Escherichia coli strains, e.g., E. coli K-12 J53-1, were not able to utilize citrate specified by the H plasmids. Most E. coli strains harboring the multicopy Cit+ plasmid pOH2, a derivative of pBR322, required only 18 to 24 h to express the Cit+ phenotype, but E. coli J53-1 (pOH2) required at least 72 h for expression. PMID- 3024565 TI - The ultrastructure of adrenocortical (interrenal) cells of normal and ACTH treated chick embryos. AB - The ultrastructural features of the interrenal cells have been studied in 20-days old chick embryos following the administration of adrenocorticotropin (ACTH), on the 8th or 15th day of incubation. The interrenal cells of normal embryos contain more SER than RER, mitochondria with tubular cristae, lipid droplets and Golgi complex. Adjacent cells had numerous regions of pentalaminar fusion and intermediate junctions. Neither a precise organization of medullary tissue in relation to interrenal tissue nor any structural differences between interrenal cells were found. Changes in the fine structure of the interrenal cells following ACTH treatment were extensive. The organelles that are known to be involved in the biosynthesis of steroids displayed structural modifications. These were mainly SER, mitochondria, and lipid droplets. Changes were also noticeable in the Golgi complexes, membrane-bound dense bodies (especially in 15 days treated embryos). These results indicate the well developed organelles in the interrenal cells of 20-days-old embryos, and their capacity to respond to ACTH stimulation as early as the 8th day of embryogenesis. PMID- 3024566 TI - Oxidative inactivation of the calcium-stimulated neutral proteinase from human red blood cells by divicine and intracellular protection by reduced glutathione. AB - Calpain, the micromolar Ca2+-requiring form of Ca2+-stimulated neutral proteinase purified from human red cells, is remarkably inactivated during autoxidation of divicine (2,6-diamino-4,5-dihydroxypyrimidine), an aglycone implicated in the pathogenesis of favism. Inactivation of purified calpain is produced, in decreasing order of efficiency, by transient, probably semiquinonic species arising from autoxidation of divicine, by the H2O2 that is formed upon autoxidation itself, and by quinonic divicine, respectively. Purified procalpain, the millimolar Ca2+-requiring form that can be converted to the fully active calpain form by a variety of mechanisms, is less susceptible than calpain itself to inactivation by the same by-products of divicine autoxidation. When intact red cells are exposed to autoxidizing divicine, procalpain undergoes a significant loss of activity. At 1 mM divicine, intracellular inactivation is observed with procalpain only, while the activity of a number of red cell enzymes is unaffected. Inactivation of procalpain is consistently greater in red cells from glucose-6-phosphate dehydrogenase-deficient subjects than in normal cells. Restoration of normal levels of glucose-6-phosphate dehydrogenase activity by means of entrapment of homogeneous human glucose-6-phosphate dehydrogenase in the deficient red cells results in normal stability of intracellular reduced glutathione; decreased susceptibility of procalpain to inactivation by autoxidizing divicine. These findings suggest that in the glucose-6-phosphate dehydrogenase-deficient red cells the procalpain-calpain system is a major target of divicine cytotoxicity. PMID- 3024567 TI - Hepatic 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase: phosphate dependence and effects of other oxyanions. AB - The effects of various oxyanions on the activities of rat liver 6-phosphofructo-2 kinase/fructose-2,6-bisphosphatase (EC 2.7.1.105/3.1.3.46) were examined. No evidence could be found for an absolute dependence of the kinase activity on inorganic phosphate as was recently reported by M. Laloux, E. Van Schaftingen, and H.-G. Hers ((1985) Eur. J. Biochem. 148, 155-159). Rather, phosphate and arsenate activated the kinase by decreasing the enzyme's Km for fructose 6 phosphate without affecting its Km for ATP or Vmax. The Km of the kinase for fructose 6-phosphate in the presence of inorganic phosphate was found to be significantly lower (6 microM) than previously reported (30 microM) when the hydrolysis of fructose 2,6-bisphosphate by the concomitant bisphosphatase activity at low Fru 6-P concentrations was taken into account. The KA's for phosphate and arsenate activation of the kinase were 0.2 and 0.3 mM, respectively. A number of other oxyanions, including pyrophosphate, sulfate, tungstate, selenate, and molybdate all inhibited the kinase by increasing the Km for fructose 6-phosphate. The apparent Ki's for inhibition of the kinase were in the 0.5-1 mM range. In contrast, all of these oxyanions activated the bisphosphatase, with half-maximal effects requiring millimolar concentrations. Inorganic phosphate was the most potent activator with a KA of 1 mM. In contrast to the other oxyanions, vanadate and meta-periodate inhibited the kinase but had no effect on the bisphosphatase. Vanadate appeared to be a noncompetitive inhibitor since its effects were not overcome by Pi, ATP, or fructose 6 phosphate, and the species responsible was shown to be decavanadate. Like vanadate, meta-periodate had no effect on the bisphosphatase, though it was a potent inhibitor (I0.5 = 30 microM) of the kinase. Its effects were shown to be time-dependent and reversed by dithiothreitol, suggesting that it acted by an oxidative mechanism. These results augment the mounting body of evidence that the enzyme's two reactions are catalyzed at discrete active sites. PMID- 3024568 TI - A dose-response study of forskolin, stimulatory hormone, and guanosine triphosphate analog on adenylate cyclase from several sources. AB - We have described relationships involving forskolin stimulation of adenylate cyclase (AC) from a variety of sources and the potentiation of forskolin effects by stimulatory hormones (glucagon, ACTH, and epinephrine) and beta, gamma imidoguanosine 5'-triphosphate (Gpp(NH)p). The effects on AC were examined using membrane preparations of rabbit adipocytes, rat adipocytes, rat erythrocytes, and rat liver. Also examined was the AC of liver membranes of rat pretreated with pertussis toxin as well as that solubilized from rat liver membranes. Maximal forskolin stimulation of AC in all preparations studied revealed a consistent 10 fold increase in AC activity. The EC50 for forskolin was 10 microM for rat liver, 15 microM for rabbit and rat adipocytes and 17 microM for rat erythrocyte AC stimulation. In all cases the AC activity attained by forskolin stimulation was further enhanced by stimulatory hormones in a dose-dependent manner. Furthermore, a combination of all three activators (forskolin, stimulatory hormone, and Gpp(NH)p) resulted in an even greater overall stimulation to levels ranging from 25- to 30-fold over unstimulated activity levels. In the presence of saturating levels of each stimulatory hormone and Gpp(NH)p, the EC50 for forskolin diminished markedly to the range of 0.5 to 4.0 microM. In the absence of any apparent tissue specificity for forskolin stimulation, the general pattern of these results further implicates the catalytic site of the AC complex as the site of forskolin activation. Furthermore, activation of additional components of the complex by Gpp(NH)p and tissue specific hormones may further influence the AC activity and thereby potentiate the stimulation by forskolin. PMID- 3024569 TI - Conditional inhibition of forskolin-activated adenylate cyclase by guanosine diphosphate and its analog. AB - Forskolin-activated adenylate cyclases (AC) in intact membranes, solubilized with Lubrol or eluted following adsorption on a forskolin-Sepharose column, were examined for inhibition by GDP and GDP beta S. AC in intact membranes of rat or rabbit adipocytes was activated by 100 microM forskolin and further potentiated by 10 microM Gpp(NH)p in combination with either 230 microM epinephrine or 50 mU X ml-1 ACTH. GDP (0-1 mM) or GDP beta S (0-500 microM) inhibited activation in a dose-dependent manner to a level similar to or slightly below that produced by 100 microM forskolin alone. Forskolin at 100 microM stimulated solubilized AC of rabbit adipocytes and rat liver membranes for 10 +/- 4 to 160 +/- 10 and from 26 +/- 2 to 274 +/- 21 pmol(mg X min)-1, respectively, in the absence of GDP beta S; forskolin-activated activity decreased from 160 +/- 10 to 157 +/- 6 and from 274 +/- 21 to 238 +/- 14 pmol(mg X min)-1 in the presence of 500 microM GDP beta S. Forskolin-activated solubilized enzyme was further potentiated by 10 microM Gpp(NH)p from 160 +/- 10 to 289 +/- 52 and from 274 +/- 21 to 702 +/- 50 pmol(mg X min)-1. GDP beta S at 500 microM inhibited 93 and 103% of the Gpp(NH)p potentiated activity. AC of rat adipocytes eluted from forskolin-Sepharose affinity column with 500 mM NaCl and 100 microM forskolin was not significantly activated by Gpp(NH)p nor inhibited by GDP beta S. However, it was activated by forskolin. The lack of inhibition of unmodified forskolin-activated activity by GDP or GDP beta S in contrast to the inhibition of Gpp(NH)p-activated enzyme or Gpp(NH)p-potentiated forskolin-activated enzyme may be a general phenomenon descriptive of the action of forskolin on AC. Furthermore, inhibition of forskolin-activated AC by GDP and its analog may be a useful index in analyzing the degree of guanine nucleotide potentiation of this enzyme. PMID- 3024570 TI - Carbon-centered free radical intermediates in the hematin- and ram seminal vesicle-catalyzed decomposition of fatty acid hydroperoxides. AB - Heme-catalyzed decomposition of unsaturated hydroperoxy fatty acids has been proposed to proceed via carbon-centered free radicals (delocalized at positions C11, C12, and C13 for 15-hydroperoxy-eicosatetraenoic acid (15-HPETE). The stable products are usually epoxy fatty acids and epoxy alcohols. Hydroperoxides from arachidonic acid can decompose via this mechanism to form leukotrienes of potential biological significance and can catalyze the epoxidation of proximal carcinogens to ultimate carcinogenic metabolites. We have used electron spin resonance spin-trapping techniques to detect carbon-centered radicals formed by heme- or ram seminal vesicle-catalyzed decomposition of 15-HPETE. For both systems we detect both a short- and a long-lived radical adduct. We proposed that these radical adducts are derived from C11 and C13 carbon-centered free radicals generated in the decomposition of 15-HPETE. PMID- 3024571 TI - Partial purification and characterization of the interconvertible forms of trehalase from Saccharomyces cerevisiae. AB - Cryptic trehalase from Saccharomyces cerevisiae was purified about 3000-fold. The recovery of 970% of the original "activity" indicated the removal of an inhibitor of the enzyme. Active trehalase, obtained through phosphorylation of cryptic trehalase by cAMP-dependent protein kinase, was isolated by chromatography on DEAE-cellulose. A major phosphorylated protein, with an apparent Mr of 86,000, was detected after SDS-polyacrylamide gel electrophoresis. This protein band correlated exactly with the elution profile of trehalase activity and 32Pi incorporation into the enzyme on DEAE-cellulose chromatography. Partially purified active trehalase showed absolute specificity towards trehalose with an apparent Km of 4.79 X 10(-3) M. Both forms of the enzyme showed an apparent molecular weight of 160,000, by gel filtration. Centrifugation on a glycerol density gradient indicated multiple forms of trehalase-c, with Mr of 320,000, 160,000, and 80,000. After activation of each of these forms by protein kinase, a single form of trehalase-a was observed, with a Mr of 160,000. Trehalase-c appears to be a totally inactive form of the enzyme. The only mechanism of activation seems to be phosphorylation by cAMP-dependent protein kinase. When the protein kinase concentration was varied, at a fixed trehalase-c concentration, a sigmoidal activation plot was obtained. This result suggests the occurrence of multiple forms of cryptic trehalase. PMID- 3024572 TI - Regulation of 25-hydroxyvitamin D3 metabolism in a human promyelocytic leukemia cell line (HL-60): 1,25-dihydroxyvitamin D3 stimulates the synthesis of 24,25 dihydroxyvitamin D3. AB - The human promyelocytic leukemia cell line HL-60 undergoes macrophage-like differentiation after exposure to 1,25-dihydroxyvitamin D3 [1,25(OH)2D3], the biologically active metabolite of vitamin D3. In the current study, we demonstrate that 1,25(OH)2D3 also regulates 25-hydroxyvitamin D3 [25(OH)D3] metabolism in HL-60 cells. The presence of 1,25(OH)2D3 in the culture medium of HL-60 cells stimulated the conversion of 7-10% of the substrate [25(OH)D3] to a more polar metabolite, which was identified as 24,25-dihydroxyvitamin D3 [24,25(OH)2D3] from the elution positions on sequential HPLC systems and the sensitivity to periodate treatment. The HL-60 subclone HL-60 blast, which is unresponsive to 1,25(OH)2D3 in terms of differentiation, also responded to 1,25(OH)2D3 treatment with the production of 24,25(OH)2D3. Maximal stimulation of 24,25(OH)2D3-synthesis (approximately 7 pmol/5 X 10(6) cells) in HL-60 cells was noted with a 12-h exposure to 10(-9) M 1,25(OH)2D3. The ability of vitamin D3 metabolites other than 1,25(OH)2D3 to induce the synthesis of 24,25(OH)2D3 in HL 60 cells was, with the exception of 1 alpha-hydroxyvitamin D3, in correlation with their reported affinities for the specific 1,25(OH)2D3 receptor which is present in HL-60 cells. Treatment of HL-60 cells with phorbol diesters abolished the 1,25(OH)2D3 responsiveness, while treatment with dimethylsulfoxide and interferon gamma did not markedly alter the 25(OH)D3 metabolism of HL-60 cells. Small amounts (approximately 1% of substrate) of two 25(OH)D3 metabolites, which comigrated with 5(E)- and 5(Z)-19-nor-10-keto-25-hydroxyvitamin D3 on two HPLC solvent systems, were synthesized by HL-60 cells, independently from 1,25(OH)2D3 treatment or stage of cell differentiation. Our results indicate that 1,25(OH)2D3 influences 25(OH)D3 metabolism of HL-60 cells independently from its effects on cell differentiation. PMID- 3024573 TI - Structure of monkey kidney cell RNA polymerase II: characterization of RNA polymerase associated with SV40 late transcriptional complexes. AB - Three subspecies of RNA polymerase II have been described in eucaryotic cells and designated IIO, IIA, and IIB. Although their relative proportions vary among different sources, RNA polymerases IIA and IIB constitute the bulk of most purified RNA polymerase II preparations. Antibodies against calf thymus RNA polymerase II were used to estimate the amount of polymerase II subspecies in monkey kidney cells, isolated nuclei, and SV40 late transcriptional complexes. We have found that RNA polymerase IIO is present in whole cells and isolated nuclei in higher proportions than previously reported. Subspecies IIO was found associated with SV40 minichromosomes engaged in transcription during late lytic infection. The observation that RNA polymerase IIO is associated with the cellular chromatin and SV40 minichromosomes suggest that this form of the enzyme is the subspecies active in in vivo transcription. PMID- 3024574 TI - Cytochrome b is necessary for the effective processing of core protein I and the iron-sulfur protein of complex III in the mitochondria. AB - The effect of cytochrome b on the assembly of the subunits of complex III into the inner mitochondrial membrane has been studied in a mutant of yeast (W-267, Box 6-2) that lacks a spectrally detectable cytochrome b and synthesizes a shortened form of apocytochrome b. We recently reported that several cytochrome b deficient mutants contained significantly diminished amounts of core proteins I and II as well as the iron-sulfur protein, but contained equal amounts of cytochrome c1 compared to the wild type (K. Sen and D. S. Beattie, Arch. Biochem. Biophys. 242, 393-401, 1985). In the present study, the time course of processing of precursors of both core protein I and the iron-sulfur protein which had accumulated in cells treated with the uncoupler carbonyl m-chlorophenyl hydrazone (CCCP) was noted to be significantly lower in the mutant compared to the wild type. The amounts of the mature forms of these proteins in mitochondria pulse labeled under different conditions was also considerably decreased at all times studied. The synthesis of both proteins appeared to be unaffected in the mutant, as the precursor forms of both proteins accumulated to the same extent when processing in vivo was blocked by CCCP. Furthermore, translation of RNA in a reticulocyte lysate in vitro indicated that the messenger RNAs for both proteins were present in the mutant and translated with equal efficiency. The import into isolated mitochondria of the precursor forms of the iron-sulfur protein synthesized in the cell-free system was also decreased in the mutant mitochondria. In addition, the precursor form was bound to the exterior of the mitochondrial membrane where it was sensitive to digestion with proteases. By contrast, the synthesis and processing of cytochrome c1 appeared to be unaffected in these mutants. These results suggest that cytochrome b is necessary for the proper processing and assembly of both core protein I and the iron-sulfur protein, but not for cytochrome c1, into complex III of the inner mitochondrial membrane. PMID- 3024575 TI - Inability of glucagon to regulate glycogen metabolism in rat hepatocytes isolated after fasting and refeeding high-carbohydrate diets. AB - Studies are described which demonstrate that the ability of glucagon, epinephrine, and dibutyryl-cAMP to stimulate glycogenolysis is impaired in rat hepatocytes isolated from animals starved for 24 h and then refed a sucrose-rich diet or refed standard rat chow. The impaired regulation of glycogenolysis by glucagon was observed within 24 h after refeeding and persisted for at least 3 days. The inability of glucagon to stimulate glycogen breakdown in the refed condition appeared to be due to a suppressed activation of glycogen phosphorylase and phosphorylase b kinase by the hormone. The capacity of glucagon to regulate pyruvate kinase and glycolysis was not altered by refeeding, suggesting that the defect lies beyond interaction of the hormone at its receptor. Prolonged incubation of hepatocytes from refed rats was accompanied by depletion of glycogen reserves and was accompanied by restoration of hormonal stimulation of glycogenolysis. Addition of glycogen to cell-free extracts was found to inhibit phosphorylase b kinase but not phosphorylase. The findings of this investigation are consistent with the interpretation that high levels of glycogen present of liver after refeeding may lead to a diminished activity of phosphorylase b kinase and its hormonal regulation. PMID- 3024576 TI - A possible mechanism of induction and translocation into blood stream of rat alkaline phosphatase activity by bile duct ligation. AB - We investigated the effects of bile duct ligation on alkaline phosphatase (ALP) activities in liver, calvarium, duodenum, and ileum in rats and its possible mechanism of action. ALP isozyme activities in the ligated rats were significantly elevated in the liver and duodenum, while those in the ileum and calvarium were markedly decreased. The ALP isozyme activity elevated by the ligation was obviously suppressed by prior administration of indomethacin, an inhibitor of prostaglandin synthesis. Moreover, phorbol ester also elevated the ALP activity as well as the phosphatase level in the ligated rat. However, other drugs, such as an inhibitor of protein kinase C and calmodulin, showed different effects: calmodulin stimulated an 11.0-, 1.3-, or 1.5-fold increase in ALP activity in the ileum, duodenum, or calvarium, respectively; whereas the hepatic enzyme activity was not affected. The induction by calmodulin was markedly different from that by the ligation. Moreover, imipramine, an inhibitor of protein kinase C, had little effect. These results suggest that prostaglandin is a possible ALP inducer in ligated rats, probably working by elevating the cAMP level. On the other hand, the ligation induced simultaneously de novo synthesis of the membranous and soluble ALP isozymes; and the release rate of the soluble enzyme was greater than that of the membranous isozymes, indicating that the soluble enzyme might be a main source of the induced serum ALP. Lectin affinity chromatography indicated that the soluble enzyme or induced serum enzyme may contain more fucose than that of the membranous one, suggesting that the sugar moiety in the ALP molecule may relate to the clearance of ALP from or its release into the circulation. PMID- 3024577 TI - Cooperative effects of Ca2+ and Sr2+ on sarcoplasmic reticulum adenosine triphosphatase. AB - The intrinsic fluorescence of purified Ca-ATPase from skeletal sarcoplasmic reticulum was measured in the presence of various concentrations of Ca2+, Sr2+, and Ba2+. Ca2+ and Sr2+ induce positive cooperative fluorescence enhancement, whereas Ba2+ does not change the fluorescence of ATPase. ATP does not seem to modify the kinetic parameters of Ca2+ and Sr2+ binding to ATPase. Nevertheless, p nitrophenylphosphate hydrolysis, activated by Ca2+ or Sr2+ at various pHs, changes the affinity and the cooperative behavior for both cations and two components appear in the Hill plots. For Ca2+, nH of 1.6 to 3.5 were obtained, and 1.06 to 1.83 for Sr2+; nH changes of the second component seem to be pH dependent. Differences in the ratio between rates of Ca2+ transport and substrate hydrolysis by sarcoplasmic reticulum were found, i.e., two for ATP and one for p nitrophenylphosphate. For Sr2+ this ratio was one for either ATP or p nitrophenylphosphate. PMID- 3024578 TI - [Regulation of cell growth by retinoids and gene expression]. AB - Retinoids are compounds that can elicit specific biological responses by virtue of their binding to and activating a specific receptor or a set of receptors. Retinoids produce various specific biological effects, including induction of terminal differentiation, regulation of cell proliferation, regulation of gene expression and regulation of the activity of specific enzymes in cells. In this article, the effects of retinoids on gene expression are reviewed. Among these effects suppression of myc expression and induction of EGF-receptor mRNA expression are considered to be closely related to regulation of cell proliferation. The effects of retinoids on cell growth are discussed on the basis of these two actions: myc mRNA suppression and EGFR mRNA induction. The mode of retinoidal action seems to be similar to that of steroids, as many investigators suggest. The molecular mechanism of retinoidal action is considered to be the formation of a retinoid-receptor complex and its interaction with regulatory elements of DNA. The possibility of application of the methodology used in the investigation of steroidal action to the study of retinoidal action is also discussed. PMID- 3024579 TI - [Phase II study of bronchial artery infusion of mitomycin C in non-small cell lung cancer]. AB - A phase II study of bronchial artery infusion of mitomycin C (MMC) was performed in 14 patients with non-small cell lung cancer (6 patients with adenocarcinoma, 6 patients with squamous cell carcinoma and 2 patients with large cell carcinoma). MMC at a dose of 20 mg was infused into the bronchial artery (total dose 20-60 mg, mean 27 mg). Among the 14 patients, one with adenocarcinoma of the lung showed partial response. The response rate for bronchial artery infusion of MMC was thus 7.1%. The toxic effects included anemia (35.7%), leukopenia (28.6%), thrombopenia (14.3%), elevation of GPT (14.3%), anorexia (14.3%), nausea (7.1%) and eruption (7.1%). PMID- 3024580 TI - [Chemoembolization therapy with cisplatin.lipiodol (CDDP.lipiodol) in primary liver cancer--with special reference to hepatocellular carcinoma]. AB - From Jan., 1985 to Mar., 1986 thirty-six patients with primary liver cancer received transcatheter arterial chemoembolization therapy with Cisplatin (100 mg) blended into Lipiodol (5 ml) and simple embolization therapy with Gelfoam particles. Thirty-three cases out of 36 had hepatocellular carcinoma, one had hepatoblastoma and one had adenocarcinoma. Ten (31%) out of 32 had hepatocellular carcinoma, and showed objective tumor reduction greater than 50% (partial response) regarding the main tumor. Of the 33 there was one sudden death due to intracerebral hemorrhage. Only two out of 25 cases with daughter nodules showed slight reduction. Almost all cases with daughter nodules showed no response to chemoembolization therapy. Five patients died after chemoembolization therapy during the fifteen-month study period. Two patients died of liver abscess or cholecystitis and surrounding abscess, one died of intracerebral hemorrhage, one died of hepatic failure and the remaining case was one of tumor death. PMID- 3024581 TI - [A phase I study of recombinant human tumor necrosis factor (rHu-TNF: PT-050). The PT-050 Study Group]. AB - A Phase I study of rHu-TNF (PT-050) was conducted in patients with various malignant tumors refractory to conventional therapy. rHu-TNF was administered by 30-min intravenous (i.v.) infusion or intratumor (i.t.) injection. The starting dose of 1 X 10(5) U/body was increased to 5 X 10(6) U/body in the i.v. group and to 2 X 10(6) U/body in the i.t. group. rHu-TNF was evaluated in 41 patients among the enrolled 43 patients of the i.v. group, and in 9 out of 10 in the i.t. group. In the i.v. group, fever (68.3%), chills (75.6%), hypotension (46.3%), general fatigue (34.1%), nausea/vomiting (22.0%/22.0%), pain in the extremities (17.1%), etc. were observed as adverse reactions (ADRs), and elevation of GOT/GPT (46.3%/43.9%), elevation of ALP(26.8%)and decrease in platelets (12.2%), etc. were observed as abnormal laboratory findings. Among these, hypotension was recognized as the dose-limiting factor and the maximum tolerated dose was considered to be 1 X 10(6) U/body. Plasma levels of rHu-TNF after 30-min i.v. administration were dose-related, and decreased with half-lives of 0.5-2.4 hours. In the i.t. group, ADRs occurred with a lower incidence than in the i.v. group except for fever, chills and general fatigue. Plasma levels after i.t. administration were all within the assay limit. Evident tissue necrosis was observed in the region where rHu-TNF was administered in the i.t. group. PMID- 3024582 TI - [Combination therapy of high-dose carboquone and OK-432 in unresectable non-small cell lung cancer]. AB - A combination of OK-432 and high-dose Carboquone (12-22 mg/m2) was administered to 17 patients with unresectable non-small cell lung cancer. The response rate was 42.9% (CR-1, PR-5) among 14 patients in whom measurement of tumor diameter was possible. With regard to hematologic adverse effects, 13 patients had a WBC count of less than 3,000, and 6 patients had a platelet count of less than 50,000. Duration of WBC nadir was not longer than 3 days, and there were no cases of infectious complication or bleeding tendency. Other side effects were all transient. PMID- 3024583 TI - [Chemotherapy of balloon-occluded arterial infusion in a patient with unresectable hepatocellular carcinoma and autoimmune hemolytic anemia]. AB - The patient, a 76-year-old woman, was found to have a tumor in the epigastrium in April 1983 and was admitted to our hospital. She was diagnosed as having hepatocellular carcinoma in the left lobe of the liver with intrahepatic metastases in the right lobe. The patient also had autoimmune hemolytic anemia. Because of this condition and the metastases, we decided that the tumor was not resectable. Transcatheter arterial embolization was unsuccessful, and therefore, beginning on June 28, 1983, the patient was treated three times using balloon occluded arterial infusion of 10 mg of mitomycin C and 30 mg of adriamycin into the proper hepatic artery. After these treatments, the serum alpha-fetoprotein level returned to normal levels. CT scans and hepatic angiography showed that the main tumor and the metastases had become smaller. The patient presently shows no evidence of disease, three years after treatment. PMID- 3024584 TI - [Intraperitoneal administration of beta-interferon in gastric cancer patients with peritoneal recurrence (ascites and serum IFN levels and its antitumor effect)]. PMID- 3024585 TI - Characteristic immunologic profile of large atypical cells in lymphomatoid papulosis. Possible implications for histogenesis and relationship to other diseases. PMID- 3024586 TI - Lymphomatoid papulosis. Histologic and immunohistochemical studies in a patient with a scaly pigmented eruption. AB - A patient with lymphomatoid papulosis type A showed a peculiar, scaly pigmented eruption on the broad skin areas as well as papulonodular lesions. Large atypical cells characteristic of the infiltrate of the disease were observed not only in the dermal infiltrate of papular lesions, but also in the perivascular infiltrate of the scaly pigmented lesions, indicating that the latter was one of the skin manifestations of lymphomatoid papulosis. Immunohistochemical studies showed that these atypical cells expressed Ki-1 and cellular activation-associated antigens such as HLA-DR, Tac, and T9, but were not reactive with T-cell specific antibodies Leu-1, Leu-3a, and Leu-2a. PMID- 3024587 TI - Reduction of lipoxygenase products in psoriatic skin homogenates by QA 208-199. PMID- 3024588 TI - Enzyme activities involved in connective tissue metabolism in the skin of tight skin (TSK) mice. PMID- 3024589 TI - Localization of ultrastructural alterations induced in rat liver by dietary polybromobiphenyls (FireMaster BP-6). PMID- 3024591 TI - A morphologic study of cells from a human malignant fibrous histiocytoma "in situ" and cultured "in vitro". PMID- 3024590 TI - Decreased phagocytosis and superoxide anion production in alveolar macrophages of rats exposed to nitrogen dioxide. PMID- 3024592 TI - Human parvovirus infection in early rheumatoid and inflammatory arthritis. AB - Evidence of recent infection with human parvovirus B19 (HPV) was found in two patients with early rheumatoid arthritis (RA) and in four patients with acute inflammatory arthritis (IA). Both of the patients with RA but only one of the four patients with IA carried RA associated haplotypes. No evidence of persistent infection with HPV was found, but evidence of past infection with HPV was significantly more common in patients with RA than in controls. The results confirm the arthritogenic potential of HPV and are consistent with the hypothesis that rheumatoid arthritis may develop in a genetically predisposed patient after an arthritogenic insult such as an HPV infection. PMID- 3024593 TI - Effect of treatment on erythrocyte phosphoribosyl pyrophosphate synthetase and glutathione reductase activity in patients with primary gout. AB - The activities of erythrocyte phosphoribosyl pyrophosphate (PRPP) synthetase and glutathione reductase (GTR) were studied in 26 patients with primary gout who were receiving no treatment or treatment with either allopurinol or azapropazone, and compared with the activity in a group of healthy controls. The activity of PRPP synthetase was significantly higher in the gout group and was not influenced by either drug. No significant difference in the activity of GTR was observed. The failure of either drug to suppress the increased activity of PRPP synthetase associated with gout is discussed. PMID- 3024594 TI - Sexual function in women after proctocolectomy. AB - One hundred women who had undergone proctocolectomy with a continence-preserving procedure (50 Kock pouches, 50 ileoanal anastomoses) for ulcerative colitis or polyposis coli were interviewed regarding their preoperative and postoperative sexual function. Frequency of intercourse increased and the incidence of dyspareunia decreased after operation in both groups. Patients who had a Kock pouch had a greater incidence of persistent postoperative dyspareunia than patients who underwent an ileoanal procedure (38% vs. 18%, p less than 0.02). Only one patient in each group reported a postoperative disturbance in ability to achieve orgasm. Most women reported no change in their menstrual cycle, but patients with a Kock pouch had more episodic vaginal discharge than patients with an ileoanal anastomosis (18% vs. 0%, p less than 0.001). Postoperative fertility was minimally impaired. Overall, the majority of women in this study who underwent proctocolectomy for benign diseases experienced enhanced sexual function after operation, which they attributed mainly to improved health. PMID- 3024595 TI - Pylorus-preserving pancreatoduodenectomy. A clinical and physiologic appraisal. AB - Since 1978, 252 patients from different centers in the world have undergone pylorus-preserving pancreatoduodenectomy. Fifty-five per cent of the patients had malignant tumors in the region of the head of the pancreas. The overall operative mortality rate was 2.8%. Anastomotic leakage and fistulae occurred in 19% of the patients. Pancreatic, biliary, and enteric fistulae represented 11%, 4%, and 4%, respectively. Peptic ulcers were subsequently diagnosed in seven patients (3%), two of whom required vagotomy and antrectomy. Delayed recovery of gastric function was the most common complication of this operation, with an overall incidence of 30%. Although the cause of this gastric dysfunction is unknown, its transient nature in most patients makes expectant therapy with gastric tube drainage the best remedy when the problem is encountered. Pylorus-preserving pancreatoduodenectomy decreased the incidence of postgastric surgery syndromes that are commonly associated with the standard Whipple operation. The existing data support the continued use of the operation and the need for future laboratory and clinical investigation of its physiologic impact. PMID- 3024596 TI - Intracardiac extension of Wilms' tumor. A report of the National Wilms' Tumor Study. AB - Extension of Wilms' tumor through the inferior vena cava into the heart presents a formidable clinical challenge. Excision of such a tumor without provoking emobilization may require cardiopulmonary bypass (CPB). The completeness of excision and the likelihood of tumor embolization during operation guide subsequent radiation therapy (RT) and chemotherapy. To help define these issues, the clinical records of 15 patients enrolled in three National Wilms' Tumor Studies (NWTS) who had intracardiac tumor extension (ICE) were reviewed. The median age at diagnosis was 4 years. One patient had clear cell sarcoma (CCS); the remainder had favorable histologic findings (FH). The clinicopathologic stage was stage II in one patient, stage III in eight patients, and stage IV in six patients. ICE was detected before operation in six patients, during operation in five patients, and after operation in five patients. CPB was used in 10 patients. Eleven patients (73%) had operative complications, with major intraoperative hemorrhage occurring most often (six patients). Complications occurred less often when ICE was recognized before operation (three of six patients) than when it was not (eight of nine patients). Embolization occurred in only two patients. There were no operative deaths. The patient with CCS died. Eleven of 14 patients with FH survived, with an actuarial event-free, 2-year survival rate of 86%. There were no patients in the first NWTS. Of the six patients in the second NWTS (NWTS 2), four died (67%). All nine patients in the third NWTS (NWTS-3) survived, but follow-up was shorter (median 4 years 9 months vs. 2 years 7 months). No particular surgical procedure was associated with an increased death rate. This review suggests Wilms' tumor with ICE presents a formidable surgical undertaking but has a relatively good prognosis. Embolization is an uncommon event in ICE (two patients, 13.3%), allowing a planned operative approach. Echocardiography and ultrasonography provide accurate preoperative diagnosis. And ICE should be suspected in patients with extensive vena cava thrombosis or who have hypotension or heart failure during examination or surgery. PMID- 3024597 TI - Five-year survival in treated stage I and II small cell carcinoma of the lung. AB - Ten consecutive patients were treated more than five years ago, for small cell carcinoma of the lung in clinical and surgical stages I or II. Patients underwent initial surgical resection, followed by intensive combination chemotherapy for at least a year, or to limit of tolerance. Four patients were classified as stage II, T2 N1; 4 had T2 N0; and 2 had T1 N0. One patient (T2 N1) died of tumor recurrence in the central nervous system 14 months after resection. Two died of other causes before five years, one (T2 N0) of a pulmonary embolus on the seventh postoperative day, and the other (T2 N1) of carcinoma of the prostate at 50 months. Seven patients (70%) remained well and disease-free at five years postoperation. Two of the 7 died of unrelated causes, one (T2 N0) at 72 months and one (T2 N1) at 108 months. Five remain well at 61 to 112 months after resection. Although this series is small, no reports have shown comparable survival in a defined group of small cell carcinoma patients treated nonoperatively or by surgical resection alone. PMID- 3024598 TI - Temporal effects of diethylstilbestrol administration in vivo on testosterone production in Leydig cells. AB - The temporal nature of estrogenic suppression of Leydig cell testosterone production was investigated. Adult rats were injected SC with 50 micrograms/100 g BW of DES or vehicle. Animals were sacrificed at 4, 8, or 12 h following a single injection or at 12 h following the latter of two daily injections for 1 or 2 days. Collagenase-dispersed interstitial cells were obtained, and population I and II Leydig cells were subsequently isolated on metrizamide gradients. Population I and II Leydig cells produced in vitro testosterone levels of 7.19 +/ 0.86 and 12.84 +/- 1.86 ng/10(6) cells/3 h, respectively. These levels were increased to 10- to 13-fold in the presence of hCG of dbcAMP. No significant difference was noted in the responsiveness of these two populations to the in vitro additions. DES administration in vivo for 8-48 h resulted in dramatic and significant decreases in basal and stimulated testosterone production in vitro in both populations. However, DES treatment for 4 h was relatively ineffective in blocking testosterone production in vitro. The inhibitory patterns exhibited by the two populations differed considerably. Population I displayed a uniform degree of inhibition throughout the treatment, whereas population II exhibited a more transient suppression by estrogen. Thus, population II appeared to be less sensitive to the estrogenic effects than population I at 48, 24, and 12 h of treatment. These data indicate that both population I and population II Leydig cells become sensitive to the inhibitory effects of estrogens between 4 and 8 h of in vivo treatment and suggest that certain differences exist between the two populations with respect to the temporal action of estrogens. PMID- 3024599 TI - Influence of dihydrolysergic acid amide on serotonergic and alpha-adrenergic receptors in human blood platelets and femoral veins in vitro. AB - The effect of dihydrolysergic acid amide (DLSA) on responses of human blood platelets and isolated postmortem femoral veins to serotonin and catecholamines was studied in vitro in comparison to dihydroergotamine (DHE). DLSA inhibited the 5-HT-potentiated, ADP-induced platelet aggregation in approximately the same concentration range as DHE. It was three orders of magnitude less potent in inhibiting the adrenaline-induced aggregation and specific binding of 3H yohimbine to intact platelets than DHE. In femoral vein strips, DLSA caused an increase in tone in the same concentration range (0.01-1.0 mumol/l) as DHE; however, its efficacy was somewhat lower. At concentrations of 0.1 to 1.0 mumol/l it antagonized the 5-HT-induced contractile response in a noncompetitive manner, the antagonist potency being one order of magnitude lower than that of DHE. DLSA (1 mumol/l) did not inhibit the noradrenaline-induced venoconstriction. The results show that DLSA possesses agonist activity in femoral veins and 5-HT antagonist activity in platelets and veins. PMID- 3024600 TI - A new vasodilator 3-isobutyryl-2-isopropylpyrazolo[1,5-a]pyridine (KC-404) has a dual mechanism of action on platelet aggregation. AB - 3-Isobutyryl-2-isopropylpyrazolo [1,5-a]pyridine (KC-404) at a concentration of greater than or equal to 4.34 X 10(-5) M inhibited adenosine diphosphate-, arachidonic acid- and collagen-induced aggregation of rabbit platelets. In rabbit, KC-404 (0.5 and 2 mg/kg, i.v.) caused a decrease in weight of collagen strip extracorporeally superfused with arterial blood, because of a disaggregation of deposited platelet aggregates. This disaggregatory effect of KC 404 was markedly diminished by the pretreatment of animals with aspirin. KC-404 (greater than or equal to 4.34 X 10(-6) M) and its major metabolite diOH-KD-404 (greater than or equal to 3.78 X 10(-7) M) significantly potentiated the anti aggregatory action of prostacyclin on rabbit platelets. KC-404 (greater than or equal to (4.34 X 10(-8) M) exerted a similar effect in rat platelets. KC-404 had no significant effect on 6-keto-PGF1 alpha and thromboxane A2 formation by rat aortic segment and rabbit platelets, respectively. KC-404 inhibited cyclic AMP phosphodiesterase (Ki = 91 microM). The present results indicate that KC-404 exhibits its anti-platelet effect via the inhibition of cyclic AMP phosphodiesterase activity in platelets and via the potentiation of anti aggregatory activity of prostacyclin on platelets. PMID- 3024601 TI - Control of enzyme synthesis in the oxalurate catabolic pathway of Streptococcus faecalis ATCC 11700: evidence for the existence of a third carbamate kinase. AB - Streptococcus faecalis ATCC 11700 uses oxalurate as a sole energy source for growth. An oxamate carbamoyltransferase and a carbamate kinase, both induced by oxalurate, are involved in this process. The oxalurate-induced kinase is specific for the pathway. Its properties are different from those of the previously characterized agmatine and arginine-induced kinases. Glucose, but not arginine, nor agmatine, two other energy sources, represses the oxalurate pathway. In contrast, oxalurate was found to be at least as effective as glucose in repressing the arginine deiminase pathway in arginine grown cells, or the agmatine deiminase pathway during growth on agmatine. PMID- 3024602 TI - AK phenotypic changes observed in some hematologic diseases. AB - Determination of AK types in 372 patients of Hematologic Clinic, revealed in many cases changes in electrophoretic pattern of AK1 type namely the occurrence of an additional protein band. These changes were observed mostly in the acute granulocytic leukemia, lymphoblastic leukemia, and aplastic anemia. In the chronic granulocytic leukemia they were present as a rule, during the blastic crisis. Phenotypic changes were transient and the repeated examinations showed disappearance of an additional band in some patients. Etiology of the changes observed is still unclear. PMID- 3024603 TI - Extent of axillary dissection preceding irradiation for carcinoma of the breast. AB - "Limited" surgery and irradiation have become more popular therapeutic options for women with stage I and stage II breast cancer, and surgical attention to the axilla is part of this approach. To understand the limitations of whatever axillary procedure is recommended, we undertook a retrospective analysis of the records of 277 women who had undergone radical or modified radical mastectomy. Of this group, 127 had metastases to at least one axillary or interpectoral lymph node. Skip metastases occurred in 13% of women with positive nodes; two women (1.6%) had metastases only to level III nodes, and two women had metastases only to interpectoral nodes. The extremely uncommon occurrence of metastases to level III alone or to interpectoral nodes alone, but the greater likelihood of skip metastases to level II, argues for both level I and level II axillary dissection preceding irradiation for patients with invasive carcinomas of the breast. PMID- 3024604 TI - [Urea utilization in growing lambs. 4. N balances with unprocessed rations]. AB - N balance experiments were carried out with lambs of the ages of 8, 12 and 15 weeks fed with wheat rations with and without 2% urea supplement (N 1 and N 2) as well as with 3% urea and 20% straw (N 3) or with a soya bean meal supplement (N 4). There were no significant differences in the digestibility of the crude nutrients and in per cent of N retention between the individual ages. The straw supplement decreased the digestibility of organic matter, crude protein, crude fibre and NfE. The supplements of soya bean meal or urea increased the crude protein content in comparison to the wheat ration without supplements by 6% in the dry matter and resulted in N intakes 55 ... 60% higher and in 23 ... 38% higher N retention, which was, however, lower in relation to N intake. There were no significant differences with regard to N retention between N 2, N 3 and N 4. Consequently urea supplement to the feed mixture with 14% native crude protein resulted in increased N retention, which was not lower than with a soya bean meal supplement. PMID- 3024605 TI - Ophthalmoscopic and histologic findings in cytomegalovirus retinitis treated with BW-B759U. AB - Recent reports have suggested a role for BW-B759U in the treatment of cytomegalovirus retinitis. The present study presents the ophthalmoscopic features in three treated patients, and examines the microscopic findings in the eyes of the first patient ever treated with BW-B759U for cytomegalovirus retinitis. Ophthalmoscopic findings of improvement on drug therapy included remission of perivascular infiltration, stabilization of the extent of the retinal area involved, and development of a pigmented chorioretinal scar in the involved area. Relapses occurred with cessation of therapy, and repeated courses of therapy were required. One patient developed a rhegmatogenous detachment with retinal breaks in an atrophic area of treated retinitis. Electron microscopic examination of the eyes of one patient disclosed the persistence of cytomegalovirus inclusions in retinal cells despite previous drug therapy. These findings suggest a virostatic action for BW-B759U. PMID- 3024606 TI - Treatment of cytomegalovirus retinopathy in patients with acquired immunodeficiency syndrome. Use of the experimental drug 9-[2-hydroxy-1 (hydroxymethyl)ethoxymethyl]guanine. AB - Cytomegalovirus (CMV) retinopathy, a relentlessly progressive disease that results in permanent blindness, is the most common opportunistic infection of the eye in patients with the acquired immunodeficiency syndrome. Twenty patients with the acquired immunodeficiency syndrome with CMV retinopathy were treated with a new, experimental, antiviral drug, 9-[2-hydroxy-1 (hydroxymethyl)ethoxymethyl]guanine (BW B759U), in dosages ranging from 5.0 to 14.0 mg/kg/d for a ten- to 20-day course. In 19 patients (95%), treatment halted the progression of infection and decreased retinal opacification, hemorrhage, and vasculitis. Vision remained stable in most cases. Six patients received no additional treatment. Fourteen patients received continued treatment with a lower maintenance dosage. Retinal disease reactivated in all patients who did not receive maintenance therapy immediately after initial treatment, indicating persistence of live virus despite drug therapy. Reactivation of disease also developed in four (40%) of ten patients receiving continuous, uninterrupted maintenance therapy for longer than three weeks. Reversible neutropenia, requiring cessation of treatment, developed in five (25%) of 20 patients on initial treatment and five (36%) of 14 patients receiving maintenance therapy. Rhegmatogenous retinal detachment was a late complication in four patients. BW B759U appears to be useful in the management of CMV retinopathy by reducing or delaying visual loss. PMID- 3024607 TI - Demonstration of papillomavirus capsid antigen in human conjunctival neoplasia. AB - To investigate the association of human papillomavirus with conjunctival neoplasia, we identified 50 resected papillomas from 47 patients. Papillomas were composed of papillary or, less commonly, flat proliferations of predominantly nonkeratinizing squamous epithelium with admixed goblet cells. Koilocytosis was focally present in 30 tumors (60%). Atypia that ranged from mild to severe was present in ten lesions (20%). In addition, we examined specimens of conjunctival dysplasia or carcinoma from 61 patients. The lesions were predominantly flat proliferations of atypical epithelial cells. Twenty biopsies performed for suspected sarcoidosis were used as controls. Papillomavirus capsid antigen was demonstrated using an immunoperoxidase technique in nuclei of mature superficial epithelial cells of 23 papillomas (46%) and five dysplasias or carcinomas (8.2%) but not in the control biopsy specimens. These results suggest that papillomavirus may play a role in the etiology of conjunctival papilloma, dysplasia, and carcinoma. PMID- 3024608 TI - Genital tract papillomavirus type 6 in recurrent conjunctival papilloma. AB - An infant boy born of a mother who had condylomata (genital warts) during pregnancy and at delivery developed recurrent conjunctival papillomas and papillomas on the soft palate and the false vocal cords. A conjunctival lesion was first noticed by the mother when the infant was 4 months old and was excised and histologically diagnosed as a papilloma when he was 11 months old. The DNA sequences of genital tract human papillomavirus type 6 (HPV-6) were identified in conjunctival papilloma tissue by Southern transfer hybridization of tissue DNA extracted from a lesion excised at 29 months of age as well as by in situ hybridization of paraffin sections of the diagnostic biopsy specimen obtained at 11 months of age. It is probable that the infant acquired conjunctival infection from the mother, very likely during passage through the infected birth canal. PMID- 3024609 TI - Antigens of herpes simplex virus in whole corneal epithelial sheets from mice. AB - Mice were inoculated with herpes simplex virus in the skin of the snout or by scarification on the cornea and then examined for eye disease using a slit lamp. Whole mounts of corneal epithelium were stained for virus antigens by the peroxidase-antiperoxidase method, and infectious virus was isolated from eyewashings. Antigens were present one day after corneal inoculation, but after inoculation of the snout, there was a delay of three days before antigens were seen. This delay and the distribution of antigens were evidence of zosteriform spread from the snout to the eye via the nervous system. Disease of the cornea varied in severity and timing depending on the site of inoculation. The peroxidase-antiperoxidase method was more sensitive than isolation of virus from eyewashings and allowed the site and distribution of infected cells to be seen. PMID- 3024610 TI - Conjugative, staphylococcal plasmids carrying hitch-hiking transposons similar to Tn554: intra- and interspecies dissemination of erythromycin resistance. AB - Two staphylococcal plasmids, pWG4 and pWG25, encode production of a diffusible pigment and resistance to erythromycin and spectinomycin. The former was found occurring naturally in a clinical isolate of Staphylococcus aureus and the latter in S. epidermidis. Both plasmids are conjugative, capable of high-frequency, interspecies transfer, only isolated in the open-circular form and identical in molecular weight and pattern of restriction-endonuclease fragments. The only difference between the plasmids is in the expression of resistance, pWG4 encoding inducible and pWG25 constitutive erythromycin resistance. The resistance determinants of both plasmids behave as hitch-hiking transposons in cultural conditions that favour phage-mediated or phage-independent conjugation, always inserting a copy of themselves into the recipient's chromosome, except in S. epidermidis in which the chromosomal insertion site may be absent. The resistance determinants have been cloned and located on a 4 X 7 kbp EcoR1/HindIII restriction fragment which has a restriction map similar to that of the right arm of Tn554 (Murphy and Lofdahl, 1984). The hitch-hiking transposon of plasmid pWG25 has been designated Tn3853. PMID- 3024611 TI - Caprine arthritis-encephalitis virus infection: from recognition to eradication. PMID- 3024612 TI - The effect of colostrum-derived antibody on neo-natal transmission of caprine arthritis-encephalitis virus infection. AB - Two groups of 6 newborn goat kids were artificially fed colostrum containing antibody to caprine arthritis-encephalitis (CAE) virus, obtained from clinically affected does. Kids in group A were fed the colostrum from birth until 7 days of age, while kids in group B were fed colostrum from 1 to 3 days after birth for 7 days. Kids were fed cow's milk at all other times. Serum antibody resulting from the consumption of colostrum, detected by agar gel immunodiffusion (AGID) tests, lasted for up to 8 weeks in group A, but none was detected in group B. Four kids from each group became infected with CAE virus as demonstrated by the emergence of active immunity and by virus isolation procedures. It appeared that uptake of colostral antibody by group A did not prevent viral transmission, interfere with development of active immunity, or modify the outcome of the CAE virus infection. PMID- 3024613 TI - Caprine retrovirus infection in New South Wales: virus isolations, clinical and histopathological findings and prevalence of antibody. AB - Caprine arthritis-encephalitis virus (CAEV) was isolated by explant cultures of carpal synovial membranes and lung from 7 goats in New South Wales. These goats were clinically affected with the arthritic, neurologic, and pneumonic forms of CAEV infection either singly or in combination. CAEV antibody was detected by the gel immunodiffusion precipitin (GDP) test in 5 of the 7 goats. Serum samples from 2,708 goats, from 115 herds, were examined for CAEV antibody using the GDP test. Approximately one-third of the animals and 82% of the herds tested had CAEV antibody. The infection was common in all breeds of dairy goats with an indication of a significantly lower prevalence in the Saanen breed (24.4%) compared to Nubians, British Alpines and Toggenbergs (43.8%, 38.7% and 39.1% respectively). CAEV antibody was also demonstrated in 11 of 230 Angora goats. The infection was equally common in all age groups, with slightly higher prevalence in males (83 of 230, 36%) compared to females (648 of 2,232, 29%). Among seropositive animals 85% were clinically normal. Of 280 clinically affected goats tested only 42% had detectable antibody. One of 5 sheep that had been in contact with infected goats in one herd had CAEV serum antibody. PMID- 3024614 TI - Detection of extracellular matrix antigens (fibronectin, laminin, type IV in collagen) in paraffin embedded sections by avidin-biotin-peroxidase complex labelling. AB - An avidin-biotin-peroxidase labelling method was applied to frozen sections and to routine-fixed, paraffin-embedded sections, and compared with immunohistochemical results of stromal antigen detection (type IV collagen, laminin and fibronectin) obtained in parallel on different neoplasias. Lung, breast and gastrointestinal tract tumors were studied. Superimposable sections were obtained from cryostat and formalin-fixed, paraffin-embedded specimens (previously digested by pepsin). The data demonstrate the potential use of immunohistochemical investigation of paraffin-embedded tissues for histological analyses of tumors. PMID- 3024615 TI - Metabolic aspects of the development of experimental cardiac hypertrophy. AB - In three models of cardiac hypertrophy the significance of catecholamines and the adenylate cyclase-cyclic AMP-system was examined. Two approaches were utilized: 1. The time course of cyclic AMP alterations was correlated with the changes in adenine nucleotide and protein biosynthesis. 2. The effect of beta-receptor blockade on the obligatory increase in adenine nucleotide and protein synthesis was evaluated. In isoproterenol-elicited cardiac hypertrophy, the elevation of the cyclic AMP content was one of the earliest metabolic alterations preceding the enhancement of the biosynthesis of adenine nucleotides and proteins. beta Receptor blockade with propranolol abolished the increase in adenine nucleotide synthesis. In pressure-induced cardiac hypertrophy due to constriction of the abdominal aorta, catecholamines and the adenylate cyclase-cyclic AMP-system were found not to play a significant role. In triiodothyronine-elicited hypertrophy, the cyclic AMP level was increased very early, but beta-receptor blockade did not prevent hypertrophy nor the enhancement of cardiac adenine nucleotide biosynthesis, although the positive chronotropic and inotropic effects of triiodothyronine were abolished. This result can best be interpreted to indicate a direct effect of triiodothyronine on myocardial carbohydrate metabolism including the pentose phosphate pathway. PMID- 3024616 TI - Calcium sensitivity of myofilaments in cardiac muscle--effect of myosin phosphorylation. AB - We investigated the influence of the extent of phosphorylation of the myosin P light-chain on the calcium sensitivity of skinned heart fibres. Treatment of skinned heart fibres with PCM-phosphatase decreased the phosphorylation level of P-light-chain from 0.3 mol P/mol LC-2 and 0.2 mol P/mol LC-2* to 0.16 mol P/mol LC-2 and 0.14 mol P/mol LC-2*. Isometric tension development decreased by 34% at submaximal Ca2+-concentration (pCa 5.5) after incubation with PCM-phosphatase while tension achieved at maximum Ca2+-concentration (pCa 4.3) was not affected. The effect of desensitization on skinned fibres could be reversed by washing out the phosphatase or by addition of myosin light chain kinase. PMID- 3024617 TI - Alterations of beta-adrenoceptors subsequent to myocardial infarction. AB - Elevations and reductions of the number of beta-adrenoceptor binding sites are dependent on the strength and the duration of receptor interaction with respective agonists. In the paper presented here, results obtained by the authors concerning biosynthesis, storage and release of catecholamines following experimentally induced infarction of the myocardium in rats are compared with those of other authors for other species. Principally, storage and release of noradrenaline from ischemic hearts do not differ with the mode of inducing tissue hypoxia (stopped-flow ischemia, coronary artery ligation, occlusion of the great cardiac vein), nor for various species (rat, dog, guinea-pig). Differences are, however, present in the results of several beta-adrenoceptor binding studies performed after experimental myocardial infarction. Following acute infarction, an increase in the number of beta-adrenoceptor binding sites is generally observed, which is explained on the basis of an externalization of receptors from the cytoplasm ot the sarcolemmal membrane. Results pertaining to 2-3 days after infarction are not uniform: in guinea-pig hearts a marked drop in the number of beta-adrenoceptors has been reported, a mild rise in the number has been detected in the left and right ventricle of rat hearts. These divergent observations could arise from the experimental protocol employed, for instance in the binding assay and in the pretreatment given to the hearts prior to assay. PMID- 3024619 TI - Ligninase of Phanerochaete chrysosporium. Mechanism of its degradation of the non phenolic arylglycerol beta-aryl ether substructure of lignin. AB - This study examined the ligninase-catalysed degradation of lignin model compounds representing the arylglycerol beta-aryl ether substructure, which is the dominant one in the lignin polymer. Three dimeric model compounds were used, all methoxylated in the 3- and 4-positions of the arylglycerol ring (ring A) and having various substituents in the beta-ether-linked aromatic ring (ring B), so that competing reactions involving both rings could be compared. Studies of the products formed and the time courses of their formation showed that these model compounds are oxidized by ligninase (+ H2O2 + O2) in both ring A and ring B. The major consequence with all three model compounds is oxidation of ring A, leading primarily to cleavage between C(alpha) and C(beta) (C(alpha) being proximal to ring A), and to a lesser extent to the oxidation of the C(alpha)-hydroxy group to a carbonyl group. Such C(alpha)-oxidation deactivates ring A, leaving only ring B for attack. Studies with C(alpha)-carbonyl model compounds corresponding to the three basic model compounds revealed that oxidation of ring B leads in part to dealkoxylations (i.e. to cleavage of the glycerol beta-aryl ether bond and to demethoxylations), but that these are minor reactions in the model compounds most closely related to lignin. Evidence is also given that another consequence of oxidation of ring B in the C(alpha)-carbonyl model compounds is formation of unstable cyclohexadienone ketals, which can decompose with elimination of the beta-ether-linked aromatic ring. The mechanisms proposed for the observed reactions involve initial formation of aryl cation radicals in either ring A or ring B. The cation radical intermediate from one of the C(alpha)-carbonyl model compounds was identified by e.s.r. spectroscopy. The mechanisms are based on earlier studies showing that ligninase acts by oxidizing appropriately substituted aromatic nuclei to aryl cation radicals [Kersten, Tien, Kalyanaraman & Kirk (1985) J. Biol. Chem. 260, 2609-2612; Hammel, Tien, Kalyanaraman & Kirk (1985) J. Biol. Chem. 260, 8348-8353]. PMID- 3024618 TI - Proton translocation by cytochrome oxidase in (antimycin + myxothiazol)-treated rat liver mitochondria using ferrocyanide or hexammineruthenium as electron donor. AB - When O2 was injected into an anaerobic suspension of valinomycin-treated rat liver mitochondria inhibited with rotenone, antimycin, and myxothiazol, a small amount of O2 (0.23-0.33 ng-atom of O/mg of protein) was reduced extremely rapidly (within the 2 s time-resolution of the oxygen electrode). The subsequent steady state rate of flow of electrons to oxygen was very low [less than 3 nequiv. X s-1 X (g of mitochondrial protein)-1]. In the presence of valinomycin there was a rapid ejection of protons synchronous with the rapid phase of O2 consumption corresponding to 0.38-0.61 nequiv. of H+ X (mg of mitochondrial protein)-1. When valinomycin was replaced by carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP) there was a rapid alkalification of the medium corresponding to 0.20-0.42 nequiv. of H+ X (mg of mitochondrial protein)-1. When 2 mM-Fe(CN)6(4-) was present to re-reduce endogenous cytochrome c, O2 consumption was still biphasic but the second phase of O2 consumption was very much more rapid [600 nequiv. X s 1 X (g of protein)-1], and resulted in the virtually complete consumption of the O2 in the pulse within 4 s. With 60 microM-Ru(NH3)6(2+) as reductant, O2 consumption was even faster [1200 nequiv. X s-1 X (g of protein)-1]. In a medium containing 150 mM-choline chloride with Ru(NH3)6(2+) as reductant, the proton per reducing equivalent stoichiometry (delta H+O/e-) was +0.95 in the presence of valinomycin and -0.94 in the presence of FCCP. In choline chloride medium containing Ru(NH3)6(2+) and valinomycin, there was an uptake of K+ ions corresponding to 1.86 K+/e-. It is concluded that nearly 1 proton is translocated outwards through cytochrome oxidase per oxidizing equivalent injected in this medium. In low ionic strength sucrose-based medium, with Ru(NH3)6(2+) as reductant, delta H+O/e- was 1.05 in the presence of valinomycin, and -0.71 in the presence of FCCP. It is concluded that the translocation of protons is accompanied by net acid production in this medium. PMID- 3024620 TI - Pertussis toxin abolishes angiotensin II-induced phosphoinositide hydrolysis and prostaglandin synthesis in rat renal mesangial cells. AB - Incubation of rat renal mesangial cells with angiotensin II (0.1 microM) resulted in transient breakdown of phosphatidylinositol 4,5-bisphosphate, rapid generation of diacylglycerol and phosphatidic acid, increased 45Ca2+ influx, increased intracellular [Ca2+] as measured by quin 2, and increased prostaglandin E2 synthesis. All of these processes were markedly inhibited time- and dose dependently by prior exposure of cells to pertussis toxin. In contrast, the effects of the ionophore A23187 on 45Ca2+ influx and prostaglandin E2 synthesis were not altered by the exposure of the cells to pertussis toxin. The action of the toxin was not associated with alterations in cellular concentrations of cyclic AMP. Incubation of membrane fraction of mesangial cells with pertussis toxin resulted in ADP-ribosylation of Mr-42,000 protein. From all these results, it is likely that a G protein is involved in receptor-mediated signal transduction in renal mesangial cells. PMID- 3024621 TI - The role of Ca2+ in steroidogenesis in Leydig cells. Stimulation of intracellular free Ca2+ by lutropin (LH), luliberin (LHRH) agonist and cyclic AMP. AB - The requirements of purified rat Leydig cells for intra- and extra-cellular Ca2+ during steroidogenesis stimulated by LH (lutropin), cyclic AMP analogues and LHRH (luliberin) agonist were investigated. The intracellular Ca2+ concentrations ([Ca2+]i) were measured by using the fluorescent Ca2+ chelator quin-2. The basal [Ca2+]i was found to be 89.4 +/- 16.6 nM (mean +/- S.D., n = 25). LH, 8-bromo cyclic AMP and dibutyryl cyclic AMP increased [Ca2+]i, by 300-500 nM at the highest concentrations of each stimulator, whereas LHRH agonist only increased [Ca2+]i by a maximum of approx. 60 nM. Low concentrations of LH (less than 1 pg/ml) and all concentrations of LHRH agonist increased testosterone without detectable changes in cyclic AMP. With amounts of LH greater than 1 pg/ml, parallel increases in cyclic AMP and [Ca2+]i occurred. The steroidogenic effect of the LHRH agonist was highly dependent on extracellular Ca2+ concentration ([Ca2+]e), whereas LH effects were only decreased by 35% when [Ca2+]e was lowered from 2.5 nM to 1.1 microM. No increase in [Ca2+]i occurred with the LHRH agonist in the low-[Ca2+]e medium, whereas LH (100 ng/ml) gave an increase of 52 nM. It is concluded that [Ca2+]i can be modulated in rat Leydig cells by LH via mechanisms that are both independent of and dependent on cyclic AMP, whereas LHRH agonist action on [Ca2+]i is independent of cyclic AMP. The evidence obtained suggests that, at sub-maximal rates of testosterone production, Ca2+, rather than cyclic AMP, is the second messenger, whereas for maximum steroidogenesis both Ca2+- and cyclic-AMP-dependent pathways may be involved. PMID- 3024622 TI - The phosphatidylinositide-Ca2+ hypothesis does not apply to the steroidogenic action of corticotropin. AB - The hypothesis that ACTH (corticotropin) stimulates steroidogenesis by a mechanism that involves breakdown of polyphosphoinositides and increase in intracellular Ca2+ (called here the 'phosphatidylinositide-Ca2+ mechanism') was tested in Y-1 adrenal-tumour cells and in bovine fasciculata cells, by using incorporation of 32P and myo-[3H]inositol to study phospholipid metabolism, and quin-2 and fura 2 to measure intracellular Ca2+. As a positive control, we repeated experiments showing that angiotensin II stimulates glomerulosa cells by way of the phosphatidylinositide-Ca2+ mechanism, by using the same methods. With Y-1 and fasciculata cells, no change was observed in the incorporation of either of the labelled precursors into any phosphatidylinositide or into any of three major phosphoinositols, i.e. inositol phosphate, bisphosphate and trisphosphate. Moreover, no change in mass of any of these compounds was seen. No change was observed in the concentration of intracellular Ca2+ in Y-1 or fasciculata cells on addition of ACTH, by using either quin-2 or fura 2. By contrast, decreased incorporation of 32P into phosphatidylinositol bisphosphate and an increase in intracellular Ca2+ were seen when glomerulosa cells were treated with angiotensin II. It is concluded that the phosphatidylinositide-Ca2+ mechanism is not used by Y-1 adrenal or bovine fasciculata cells in the steroidogenic response to ACTH unless the mechanism is radically different from that seen with all other hormones so far tested in which this mechanism occurs. PMID- 3024624 TI - A computer-supported oxystat system maintaining steady-state O2 partial pressures and simultaneously monitoring O2 uptake in biological systems. AB - A feedback-controlled oxystat system is described maintaining steady-state O2 partial pressures (pO2) between 0.01 mmHg (14 nM-O2) and 150 mmHg (210 microM-O2) and simultaneously monitoring O2 uptake at rates between 0.1 and 120 microM-O2 X min-1 in suspensions of cells, in subcellular fractions and in solutions of enzymes. At pO2 values between 0.2 and 150 mmHg (0.28 and 210 microM-O2) a polarographic O2 sensor was used, and below a pO2 of 0.2 mmHg (0.28 microM-O2) the O2-dependent luminescence of the photobacterium Vibrio fischeri was utilized to monitor the actual pO2. At a selected pO2, O2 supply is maintained by injecting appropriate amounts of O2-saturated aqueous medium into the reaction chamber by using a motor-driven burette. The oxystat system is under control of a computer that reads the O2 sensors, interacts with the motor-driven burette, calculates the O2 uptake from the amounts of O2-saturated medium added, collects data from further measuring devices and provides the documentation of the results during incubation. PMID- 3024623 TI - Stimulation of phosphoinositide metabolism in hamster brown adipocytes exposed to alpha 1-adrenergic agents and its inhibition with phorbol esters. AB - The present experiments were undertaken to investigate the role of the phosphoinositides phosphatidylinositol 4-phosphate (PtdIns-4-P) and phosphatidylinositol 4,5-biphosphate (PtdIns-4,5-P2) in the alpha 1-adrenergic stimulation of respiration in isolated hamster brown adipocytes. Exposure of isolated brown adipocytes to the alpha-adrenergic-receptor agonist phenylephrine provoked a breakdown of 30-50% of the PtdIns-4-P and PtdIns-4,5-P2 after prelabelling of the cells with [32P]Pi. Coincident with the breakdown of phosphoinositides was an accumulation of labelled phosphatidic acid, which continued for the duration of the cell incubation. The time course of phosphoinositide breakdown was defined more precisely by pulse-chase experiments. Under these conditions, phenylephrine caused radioactivity in phosphatidylinositol, PtdIns-4-P and PtdIns-4,5-P2 to fall by more than 50% within 30 s and to remain at the depressed value for the duration of the incubation (10 min). This phospholipid response to alpha-adrenergic stimulation was blocked by exposure of the cells to phorbol 12-myristate 13-acetate (PMA); likewise phenylephrine stimulation of respiration was prevented by PMA. beta Adrenergic stimulation of respiration and inhibition of respiration by 2 chloroadenosine and insulin were, however, unaffected by treatment with PMA. On the assumption that PMA is acting in these cells as an activator of protein kinase C, these results suggest the selective interruption of alpha-adrenergic actions in brown adipocytes by activated protein kinase C. These findings suggest that breakdown of phosphoinositides is an early event in alpha-adrenergic stimulation of brown adipocytes which may be important for the subsequent stimulation of respiration. The results from the pulse-chase studies also suggest, however, that phenylephrine-stimulated breakdown of inositol phospholipids is a short-lived event which does not appear to persist for the entire period of exposure to the alpha 1-adrenergic ligand. PMID- 3024625 TI - Efficient entrapment of large and small compounds during vesiculation of intestinal microvilli. AB - An efficient method is described permitting the encapsulation of membrane impermeable compounds at the interior of intestinal microvilli during vesicle formation. Rat intestinal epithelial cells were isolated by high-frequency vibration and exposed transiently to iso-osmotic medium containing 5 mM-EDTA. Vesiculation of microvilli was effected by freeze-thawing instead of mechanical fragmentation or hypo-osmotic lysis. Solutes to be entrapped were mixed with the extracellular medium before freezing in liquid N2. Microvillous vesicles were isolated from thawed cell suspensions by Ca2+- or Mg2+-aggregation of contaminants and differential centrifugation. The yield, purity, orientation and transport properties of the vesicles were similar, or superior, to preparations described in the literature. A high loading efficiency was demonstrated for small impermeants (cyclic GMP, ATP, Arsenazo III) as well as proteins (albumin); in contrast, loading of isolated vesicles by hypo-osmotic shock was only partially effective (cyclic GMP, ATP) or ineffective (albumin). Entrapment of an ATP regenerating system could partially block a Mg2+-dependent conversion of intravesicular ATP into ADP. No evidence was obtained for the contribution of a proton pump to the intrinsic Mg2+-ATPase of the vesicle. Potential applications of the vesicle-loading technique in studies of brush-border transport regulation by intramicrovillar factors are discussed. PMID- 3024626 TI - The mechanism of the hormonal activation of respiration in isolated hepatocytes and its importance in the regulation of gluconeogenesis. AB - The effects of hormones on the cytochrome spectra of isolated hepatocytes were recorded under conditions of active gluconeogenesis from L-lactate. Glucagon, phenylephrine, vasopressin and valinomycin, at concentrations that caused stimulation of gluconeogenesis, increased the reduction of the components of the cytochrome bc1 complex, just as has been observed in liver mitochondria isolated from glucagon-treated rats [Halestrap (1982) Biochem. J. 204, 37-47]. The effects of glucagon and phenylephrine were additive. The time courses of the increased reduction of cytochrome c/c1 and NAD(P)H/NAD(P)+ caused by hormones, valinomycin, A23187 and ethanol were measured by dual-beam spectrophotometry and fluorescence respectively. Ethanol (14 mM) produced a substantial rise in NAD(P)H fluorescence, beta-hydroxybutyrate/acetoacetate and lactate/pyruvate ratios, no change in cytochrome c/c1 reduction, a 10% decrease in O2 consumption and a 60% decrease in gluconeogenesis. Glucagon, phenylephrine and vasopressin caused a substantial and transient rise in NAD(P)H fluorescence, but a sustained increase in cytochrome c/c1 reduction and the rates of O2 consumption and gluconeogenesis. The transience of the fluorescence response was greater in the absence of Ca2+, when the cytochrome c/c1 response also became transient. The fluorescence response was smaller and less transient, but the cytochrome c/c1 response was greater, in the presence of fatty acids. Both responses were greatly decreased by the presence of 1 mM-pent-4-enoate. Valinomycin (2.5 nM) caused a decrease in NAD(P)H fluorescence coincident with an increase in cytochrome c/c1 reduction and the rate of gluconeogenesis and O2 consumption. A23187 (7.5 mM) caused increases in both NAD(P)H fluorescence and cytochrome c/c1 reduction. The effects of hormones and valinomycin on the time courses of NAD(P)H fluorescence, cytochrome c/c1 reduction and light-scattering by hepatocytes were compared with those of 0.5 microM-Ca2+ or 1 nM-valinomycin on the same parameters of isolated liver mitochondria. It is concluded that hormones increase respiration by hepatocytes in a biphasic manner. An initial Ca2+-dependent activation of mitochondrial dehydrogenases rapidly increases the mitochondrial [NADH], which is followed by a volume-mediated stimulation of fatty acid oxidation and electron flow between NADH and cytochrome c. 10. Amytal (0.5 mM) was able to reverse the effects of hormones on the reduction of cytochromes c/c1 and the rates of gluconeogenesis and O2 consumption without significantly lowering tissue [ATP].(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3024628 TI - [Thermitase--a thermostable serine protease. VIII. Kinetic and ESR investigations on the interactions of enzymes with spin labeled peptide methyl ketones]. AB - An attempt was made to study the structure of the active center of thermitase by means of spin labeled peptide analogues. For this purpose, peptide methyl ketones SL-Alan-PheCH3 of different chain length (n = 0, 1, 2, 3; SL = 2,2,5,5 Tetramethyl-pyrrolin-1-oxyl-3-carbonyl) were synthesized. Synthesis and physico chemical properties of these compounds are described and inhibition constants Ki for the interaction of these compounds with thermitase were measured. SL-Ala2 PheCH3 and SL-Ala3-PheCH3 are strong reversible inhibitors of thermitase with Ki values of 8.9 X 10(-6) M and 4.8 X 10(-7) M, respectively, whereas the analogous compounds with n = 0 and n = 1 represent only reduced affinity for this enzyme. ESR spectra of SL-Ala2-PheCH3 and SL-Ala3-PheCH3 in the presence of thermitase reveal that a great part of the SL residues of enzyme bound inhibitor molecules is not measurably restricted in their mobility whereas another part is hindered by unspecific interaction with the protein. The kinetic and ESR data are discussed with regard to the substrate binding region of the enzyme. PMID- 3024627 TI - The effects of lithium on platelet phosphoinositide metabolism. AB - The effects on phosphoinositide metabolism of preincubation of platelets for 90 min with 10 mM-Li+ were studied. Measurements were made of [32P]phosphate labelled phosphoinositides and of [3H]inositol-labelled inositol mono-, bis- and tris-phosphate (InsP, InsP2 and InsP3). Li+ had no effect on the basal radioactivity in the phosphoinositides or in InsP2 or InsP3, but it caused a 1.8 fold increase in the basal radioactivity in InsP. Li+ caused a 4-, 3- and 2-fold enhanced thrombin-induced accumulation of label in InsP, InsP2 and InsP3 respectively. Although the elevated labelling of InsP2 and InsP3 returned to near basal values within 30-60 min, the high labelling of InsP did not decline over a period of 60 min after addition of thrombin to Li+-treated platelets, consistent with inhibition of InsP phosphatase by Li+. The effect of Li+ was not due to a shift in the thrombin dose-response relationship; increasing concentrations of thrombin enhanced the initial rate of production of radiolabelled inositol phosphates, whereas Li+ affected either a secondary production or the rate of their removal. The only observed effect of Li+ on phosphoinositide metabolism was a thrombin-induced decrease (P less than 0.05) in labelled phosphatidylinositol 4 phosphate in Li+-treated platelets; this suggests an effect on phospholipase C. Li+ enhanced (P less than 0.05) the thrombin-induced increase in labelled lysophosphatidylinositol, suggesting an effect on phospholipase A2. It is concluded that Li+ inhibits InsP phosphatase and has other effects on phosphoinositide metabolism in activated platelets. The observed effects occur too slowly to be the mechanism by which Li+ potentiates agonist-induced platelet activation. PMID- 3024629 TI - Characterization of the DNA of the hamster papovavirus. III. Mapping of inverted repeated DNA sequences within the viral genome. AB - Sequences with 2-fold axis of symmetry (inverted repeated sequences) have been detected and mapped on the hamster papovavirus (HaPV) genome by their ability to form secondary structures like hairpins or stem-loops on single-stranded HaPV DNA. DNA regions with secondary structure were visualized by electron microscopy and mapped according to the physical map of HaPV. Eleven positions containing inverted repeats could be determined. The nucleotide sequence within all of these inverted repeats may be related since these regions can crosshybridize with each other. The possible functions of these sequences within the HaPV genome are discussed with respect to conditions found for genomes like those from SV40 and polyoma virus to get first indications for the localization of DNA regions representing the origin and termination of replication of HaPV DNA. PMID- 3024630 TI - Differences in interferon-gamma response of psoriatic lymphocytes to stimulation with various mitogens. AB - Upon ConA stimulation, peripheral blood mononuclear leukocytes from psoriatic patients show an impaired IFN-gamma production. Normal IFN-gamma values were obtained, however, with PHA or PWM as inducers. Moreover, psoriatic cells responded well to vesicular stomatitis virus as inducer of IFN-alpha. Thus, the defect is not an all-out inability of all lymphocytes to produce IFN, but rather a failure to respond to weak mitogenic stimuli. Possible mechanisms are discussed. PMID- 3024631 TI - Methionine or not methionine at the beginning of a protein. PMID- 3024632 TI - Eicosanoids and aspirin in immune cell function. PMID- 3024633 TI - Oncogene homologues in yeast. PMID- 3024634 TI - Using molecular mimicry to produce anti-receptor antibodies. PMID- 3024635 TI - The role of cAMP in controlling yeast cell division. PMID- 3024637 TI - Mobile DNA elements in Drosophila melanogaster: a retrospective on serendipity. PMID- 3024636 TI - Newly discovered activities for calcitriol (1,25-dihydroxyvitamin D3): implications for future pharmacological use. PMID- 3024638 TI - Cell-surface receptors: puzzles and paradigms. PMID- 3024639 TI - Growth factors, receptors and cancer. PMID- 3024640 TI - Retroviral elements and suppressor genes in Drosophila. PMID- 3024642 TI - Additive effect of VIP or isoproterenol on forskolin-stimulated cyclic AMP accumulation in rat prostatic epithelial cells. AB - The effects of forskolin alone or in combination with vasoactive intestinal peptide (VIP) and the beta-adrenergic agonist isoproterenol on cyclic AMP accumulation in epithelial cells of rat ventral prostate were examined. Forskolin stimulated cyclic AMP in a time- and temperature-dependent manner. At 15 degrees C, forskolin behaved as a highly potent and relatively efficient stimulatory agent. The combination of forskolin with maximal doses of VIP or isoproterenol were purely additive. These results suggest that forskolin might act directly upon the catalytic subunit of adenylate cyclase in this particular class of cells. PMID- 3024641 TI - Calmodulin and ATP dependence of the high and low calcium affinity p nitrophenylphosphatase from human erythrocyte membranes. AB - In calmodulin depleted membranes from human erythrocytes, the Ca2+-dependent phosphatase showed different sensitivity to calmodulin and ATP with variable affinity towards free calcium concentrations: a calmodulin-dependent activity with high calcium affinity, K1/2 = 1.2 X 10(-7) mol/l calcium, that was fully activated at submicromolar calcium concentrations, higher concentrations being rather inhibitory; an ATP-dependent activity with lower calcium affinity, K1/2 = 10(-6) mol/l calcium, that was fully activated at 10(-5) mol/l calcium in the presence of 50-200 mumol/l ATP and was insensitive to calmodulin, and a calcium dependent phosphatase that was active at a wider ranger of free calcium, 10(-8) 10(-5) mol/l, and required the presence of both calmodulin and ATP. PMID- 3024643 TI - Theoretical and experimental study on the competition of NANA-aldolase and cytidine-5'-monophosphosialate-synthase for their common substrate N acetylneuraminic acid. AB - The competition of NANA-Aldolase and Cytidine-5'-monophosphosialate-Synthase for their common substrate NANA has been studied, with, (i) the two enzymes in solution, (ii) NANA-Aldolase in solution and Cytidine-5'-monophosphosialate Synthase immobilized in an artificial membrane (diffusion coefficient: 1.2 X 10( 3) cm2 h-1). The relation of the reaction rates for both enzyme was found 1:1 in case (i) and 2:1 in case (ii), in favor of NANA-Aldolase. These results are in agreement with the results obtained by computer simulation, where the Michaelian assumption and the diffusion effects had been considered. It was also calculated that the regulation of this branch point for the metabolic pathway of NANA is dependent on the input of NANA produced by the previous steps of the pathway and not on the concentration of CTP (second substrate of Cytidine-5' monophosphosialate-Synthase) or the parameters controlling the diffusion of NANA. Computer simulations were performed by numerical analysis. PMID- 3024644 TI - Phorbol 12-myristate 13-acetate induces beta-adrenergic receptor uncoupling and non-specific desensitization of adenylate cyclase in human mononuclear leukocytes. AB - Tumour-promoting phorbol esters such as phorbol 12-myristate 13-acetate (PMA) have been reported to modulate beta-adrenergic receptor responses in various cell types, presumably by the activation of protein kinase C. In the present investigation we assessed the effect of PMA on the beta-adrenergic receptor adenylate cyclase system of human mononuclear leukocytes (MNL). It was found that incubation of MNL with PMA resulted in a time- and concentration-dependent desensitization of isoproterenol-induced adenylate cyclase activity. However, the effect of PMA was not restricted to the beta-adrenergic receptor system, since basal adenylate cyclase activity and histamine-, prostaglandin E1-, 5' guanylylimidodiphosphate (GppNHp)-, and NaF-stimulated values were also reduced. By contrast, no effect was found on the forskolin-induced adenylate cyclase activity. The inactive phorbol ester, 4 alpha-phorbol 12,13-didecanoate had no effect on adenylate cyclase at all, suggesting that the observed PMA effect was specifically mediated by activation of protein kinase C. The reduced beta adrenergic response induced by PMA was not associated with a reduced beta adrenergic receptor number, indicating uncoupling of this receptor from adenylate cyclase. Isoproterenol competition curves for 3H-dihydroalprenolol binding to membranes from untreated and PMA-treated cells demonstrated that the uncoupling was due to a reduced ability of the agonist to promote formation of the guanine nucleotide-sensitive high affinity state of the receptor. The results indicate that PMA may cause desensitization of catecholamine-responsive adenylate cyclase in MNL, and that the major locus of alteration is the guanine nucleotide regulatory protein. PMID- 3024645 TI - Dual effects of a new hypocalcemic agent, WR-2721, on cytoplasmic Ca2+ and parathyroid hormone release of dispersed parathyroid cells from patients with hyperparathyroidism. AB - The effects of the new hypocalcemic agent, WR-2721, on calcium-regulated parathyroid hormone (PTH) release, cytoplasmic Ca2+ (Ca2+i) and membrane potential were measured in dispersed parathyroid cells from patients with hyperparathyrodisim (HPT). The drug had no effects in the absence of extracellular Ca2+ but acted synergistically with Ca2+ in the 0.5-1.5 mM range by depolarizing the cells, increasing Ca2+i and inhibiting PTH release. Although the depolarizing effect of 3.0 mM Ca2+ was unaffected by WR-2721 the drug antagonized the effect of Ca2+ by decreasing Ca2+i and stimulating PTH release. Whereas the inhibitory actions of WR-2721 on PTH release may result from the activation of the mechanism for Ca2+ gating in the parathyroid cell plasma membrane, the stimulatory effect probably reflects increased intracellular Ca2+ sequestration. The drug is considered potentially important for the treatment of HPT. PMID- 3024646 TI - A possible role for membrane lipid peroxidation in anthracycline nephrotoxicity. AB - Adriamycin causes both glomerular and tubular lesions in kidney, which can be severe enough to progress to irreversible renal failure. This drug-caused nephrotoxicity may result from the metabolic reductive activation of Adriamycin to a semiquinone free radical intermediate by oxidoreductive enzymes such as NADPH-cytochrome P-450 reductase and NADH-dehydrogenase. The drug semiquinone, in turn, autoxidizes and efficiently generates highly reactive and toxic oxyradicals. We report here that the reductive activation of Adriamycin markedly enhanced both NADPH- and NADH-dependent kidney microsomal membrane lipid peroxidation, measured as malonaldehyde by the thiobarbituric acid method. Adriamycin-enhanced kidney microsomal lipid peroxidation was diminished by the inclusion of the oxyradical scavengers, superoxide dismutase and 1,3 dimethylurea, and by the chelating agents, EDTA and diethylenetriamine pentaacetic acid (DETPAC), implicating an obligatory role for reactive oxygen species and metal ions in the peroxidation mechanism. Furthermore, the inclusion of exogenous ferric and ferrous iron salts more than doubled Adriamycin stimulated peroxidation. Lipid peroxidation was prevented by the sulfhydryl reacting agent, p-chloromercuribenzenesulfonic acid, by omitting NAD(P)H, or by heat-inactivating the kidney microsomes, indicating the requirement for active pyridine-nucleotide linked enzymes. Several analogs of Adriamycin as well as mitomycin C, drugs which are capable of oxidation-reduction cycling, greatly increased NADPH-dependent kidney microsomal peroxidation. Carminomycin and 4 demethoxydaunorubicin were noteworthy in this respect because they were three to four times as potent as Adriamycin. In isolated kidney mitochondria, Adriamycin promoted a 12-fold increase in NADH-supported (NADH-dehydrogenase-dependent) peroxidation. These observations clearly indicate that anthracyclines enhance oxyradical-mediated membrane lipid peroxidation in vitro, and suggest that peroxidation-caused damage to kidney endoplasmic reticulum and mitochondrial membranes in vivo could contribute to the development of anthracycline-caused nephrotoxicity. PMID- 3024647 TI - Deglycosylated mammalian beta 2-adrenergic receptors are still able to undergo functional coupling to Ns. AB - Mammalian beta 2-adrenergic receptors (R) have been shown to be structurally heterogeneous with respect to glycosylation (Stiles et al. J. biol. Chem. 259, 8655 (1984). They are also heterogeneous with respect to functional coupling to Ns. The ternary H.R.Ns complex can be frozen in the presence of the alkylating reagent N-ethylmaleimide. In hamster lung membranes 45% of the receptors are agonist/N-ethylmaleimide sensitive (i.e. coupling-prone receptors). beta Receptors in both native and isoproterenol/N-ethylmaleimide pretreated membrane preparations are retained by affinity chromatography on concanavalin A and wheat germ agglutinin and are equally sensitive to neuraminidase treatment. This is exhibited by the increase in mobility of the 125I-iodocyanopindolol-azide photoaffinity labeled receptor peptide in SDS-polyacrylamide gel electrophoresis. These observations suggest that there is no link between the structural and functional heterogeneity of the receptors. Moreover, both partial (using neuraminidase) and near total (using endoglycosidase F) deglycosylation of membrane-bound receptors does not affect the H.R.-Ns coupling capacity as compared to native receptors. PMID- 3024648 TI - Beta-adrenergic receptors in guinea-pig liver plasma membranes and thermal lability of [3H]dihydroalprenolol binding sites. AB - beta-Adrenergic receptors in guinea-pig liver plasma membranes were characterized by radioligand binding, using l-[3H]dihydroalprenolol ([3H]DHA), l-3 [125I]iodocyanopindolol ([125I]CYP) and dl-[3H]4-(3-tertiarybutylamino-2 hydroxypropoxy)-benzimidazole-2- one hydrochloride [( 3H]CGP-12177). The binding of both [125I]CYP and [3H]CGP-12177 to membranes exhibited high affinity (Kd = 3.5 +/- 0.2 pM for [125I]CYP and 0.75 +/- 0.10 nM for [3H]CGP-12177) and stereospecificity; the maximal binding sites were 130 +/- 15 and 137 +/- 8 fmoles/mg protein respectively. Catecholaminergic agonists competed for these binding sites in the order l-isoproterenol greater than l-epinephrine greater than l-norepinephrine, which is typical for beta 2-adrenergic receptors. The binding data are supported by parallel experiments on adenylate cyclase activation by catecholamines, and on antagonism of this activation by beta 1- and beta 2-selective blockers. The binding of [3H]DHA was excessive (Bmax = 21.4 pmoles/mg protein), exhibited low affinity (Kd = 34.6 nM), and lacked stereospecificity. When liver membranes were incubated at 50 degrees for 40 min in the presence of an agonist, l-isoproterenol, the binding of [3H]DHA to the heat-treated membranes exhibited high affinity (Kd = 1.07 +/- 0.17 nM) and the Bmax was reduced to 139 +/- 22 fmoles/mg protein. In such membranes, as opposed to native membranes, stereospecificity was evident and catecholaminergic agonists competed for the binding sites in the order typical for beta 2-adrenergic receptors. However, agonist competition of the binding to the heat-treated membranes could not be modulated by guanine nucleotides, indicating a loss of communication between the receptor and the guanine nucleotide regulatory protein. PMID- 3024649 TI - Characterization of the topoisomerase II-induced cleavage sites in the c-myc proto-oncogene. In vitro stimulation by the antitumoral intercalating drug mAMSA. AB - In an attempt to get an insight into the activity of mAMSA (a DNA topoisomerase II-mediated drug) on the human proto-oncogene c-myc, an in vitro system consisting of purified calf thymus DNA topoisomerase II and a c-myc DNA inserted in lambda phage was utilized. The occurrence of discrete bands, detected by hybridization of Southern blots with appropriate c-myc probes, indicated the presence of cleavage sites in the sole presence of DNA topoisomerase II. The band intensity increased in the presence of mAMSA, while no significant difference occurred in the cleavage pattern. The location of the cleavage sites along the c myc locus revealed a striking correspondence with that of some DNase hypersensitive sites. These results indicate that DNA topoisomerase II is most certainly implicated in the mAMSA activity and that the drug stimulates the topoisomerase II cleaving activity at specific sites, which may be involved in the biological activity of the drug. PMID- 3024650 TI - Isolation and kinetic studies of nucleoside diphosphokinase from human platelets and effects of cAMP phosphodiesterase inhibitors. AB - Nucleoside diphosphokinase (NDK) of human platelets has been purified by chromatography on Blue Sepharose CL-6B gel (purification factor of 950) and shown to be free of adenylate kinase, ATPase and adenylate cyclase. The molecular weight was 70,000 with subunits of 17,000. The pH optimum was 8.0 Km values for ATP and dTDP were determined in two ways using the pyruvate kinase-lactate dehydrogenase coupled enzyme assay. Values of 0.38 and 0.20 mM were obtained for ATP and 0.29 and 0.21 mM for dTDP. Km values for ADP (0.024 mM) and GTP (0.12 mM) were determined with the hexokinase-glucose-6-phosphate dehydrogenase coupled enzyme assay. These values are in agreement with those reported for NDK from other sources. Theophylline, which inhibits the NDK activity of intact platelets and platelet membrane preparations and inhibits the ADP-induced shape change of platelets, was shown to be a competitive inhibitor of both the free and phosphorylated forms of NDK with competitive inhibition constants (Kic) of 9.3 and 9.6 mM respectively. Papaverine, another cAMP phosphodiesterase inhibitor, which also inhibits the ADP-induced shape change of platelets, had no inhibitory effect on platelet NDK. It was concluded that the inhibitory effect of theophylline on the activity of the purified enzyme was due to the structural similarity between the methylxanthine and the adenine moiety of ADP. PMID- 3024651 TI - An antibody to dihydropyridine calcium entry blockers. A comparison with the calcium channel receptor in skeletal muscle. AB - Antibodies that recognize dihydropyridine (DHP) calcium entry blockers were elicited from rabbits. A sensitive and specific radioimmunoassay for dihydropyridines was developed and its specificity compared to the DHP binding site in skeletal muscle membranes. The antibody bound [3H]nitrendipine with a higher affinity (KD = 0.155 nM) than did the DHP receptor of skeletal muscle (KD = 1-3 mM); however, in contrast to the DHP receptor, the antibody recognized only those DHP drugs with meta-nitrophenyl substituents at the 4-position on the DHP ring. Both the antibody and receptor exhibited stereospecificity, with each site recognizing the (+)-isomer of nicardipine as the more potent. This antibody should prove useful in our studies of some potentially irreversible DHP molecules. PMID- 3024652 TI - Hydrolysis of neo-kyotorphin (Thr-Ser-Lys-Tyr-Arg) and [Met]enkephalin-Arg6-Phe7 by angiotensin-converting enzyme from monkey brain. AB - Angiotensin-converting enzyme (ACE; dipeptidyl carboxypeptidase, EC 3.4.15.1) from monkey brain was partially purified 274-fold with 4.5% yield. The optimum pH of the enzyme was 8.2, and its Km was 3.3 mM, with hippuryl-His-Leu as the substrate in 300 mM NaCl. Its molecular weight (Mr) was estimated to be approximately 260,000 by gel filtration on Sephadex G-200. On high-performance liquid chromatographic analysis, ACE hydrolyzed neo-kyotorphin Thr-Ser-Lys-Tyr Arg) with liberation of kyotorphin (Tyr-Arg), the [Met]enkephalin releaser. ACE also converted [Met]enkephalin-Arg6-Phe7 to [Met]enkephalin; then the enzyme slowly hydrolyzed the resulting [Met]enkephalin. The Km values of the enzyme for neo-kyotorphin and [Met]enkephalin-Arg6-Phe7 were 0.58 and 0.30 mM respectively. Thus, brain ACE may have a role in the formation of kyotorphin and [Met]enkephalin from their precursors but has little part in [Met]enkephalin degrading processes. PMID- 3024653 TI - Species difference in the specific receptors of platelet activating factor. AB - Relative potencies of platelet activating factor (PAF) and PAF analogs and several PAF receptor antagonists when inhibiting the [3H]PAF specific binding to human and rabbit platelet membranes and membrane fragments of human lung tissues were compared. In rabbit platelets, L-652,731 was found to be most potent in the list of PAF receptor antagonists with an equilibrium inhibition constant (Ki) of 9.83 (+/- 2.92) X 10(-9) M followed by L-653,150 greater than kadsurenone congruent to Ono-6240 greater than ginkgolide B greater than CV-3988 greater than L-651,142, whereas in human platelets the relative potencies of these PAF receptor antagonists were as follows: Ono-6240 greater than L-653,150 congruent to L-652,731 congruent to kadsurenone greater than ginkgolide B greater than CV 3988 greater than L-651,142. Ono-6240 was the most potent one with a Ki of 4.86 (+/- 1.44) X 10(-8) M which was roughly two times more potent than that in rabbit platelets, whereas the affinity of L-652,731 was about ten times less in human platelets (Ki = 1.03 (+/- 0.15) X 10(-7) M) compared to that in rabbit platelets (Ki = 9.83 (+/- 2.92) X 10(-9) M). These variations between species among PAF antagonists strongly suggest that there exists a species difference at or near the binding site of the receptor of platelet activating factor. The relative potency of these PAF receptor antagonists in human lung membranes differed very little from that in human platelets and was found to be Ono-6240 greater than L 653,150 congruent to kadsurenone congruent to L-652,731 greater than ginkgolide B greater than CV-3988 greater than L-651,142. Even though C16-PAF showed slightly higher potency in human lung, and CV-3988 and Ono-6240 showed slightly lower, the difference was too small to suggest that there is a difference in the PAF receptors between human platelets and human lung tissues. PMID- 3024654 TI - Effects of adenosine and adenosine analogues on glycogen metabolism in isolated rat hepatocytes. AB - Adenosine and adenosine analogues were incubated with isolated rat hepatocytes. Adenosine and 5'-deoxy-5'-chloroadenosine stimulated glucose release, glycogen loss, and the conversion of glycogen phosphorylase b to a. The effect was of short duration for adenosine, but of long duration for 5'-deoxy-5' chloroadenosine. The effects on glucose release and phosphorylase were blocked by theophylline, an R-receptor blocking agent, but not by nitrobenzylthioinosine or dipyridamol which are nucleoside transport inhibitors. A dose-dependent rise in cyclic AMP concentration was observed in hepatocytes 1 min after adding adenosine. It is concluded that adenosine exerts these effects in liver by activating adenylcyclase. Adenosine may be involved in the short-term regulation of hepatic glycogen phosphorylase. PMID- 3024655 TI - Inhibition of neutrophil superoxide formation by 1-(5-isoquinolinesulfonyl)-2 methylpiperazine (H-7), and inhibitor of protein kinase-C. AB - Superoxide formation of human neutrophils stimulated by phorbol 12-myristate 13 acetate (PMA), N-formyl-methionyl-leucyl-phenylalanine, or calcium ionophore A23187 was inhibited by pretreatment of the cells with 1-(5-isoquinolinesulfonyl) 2-methylpiperazine (H-7), an inhibitor of protein kinase-C, but was not inhibited by N-(2-guanidinoethyl)-5-isoquinolinesulfonamide which has a less inhibitory effect on the protein kinase-C. H-7 also inhibited superoxide formation of PMA activated cytoplasts, which lack nuclei and granules. The phosphorylation of proteins induced by PMA in the cytoplasts as well as the intact neutrophils was also inhibited by preincubation with H-7. Among several phosphoproteins affected by H-7, one protein with a molecular weight of 19,000 (pI = 4.9) was inhibited markedly. N-(2-Guanidinoethyl)-5-isoquinolinesulfonamide did not inhibit the phosphorylation of proteins induced by PMA. These findings support the possibility that the protein kinase-C is involved in the activation process of superoxide formation. PMID- 3024656 TI - Labeling of sarcoplasmic reticulum peptides with 32P-phosphate and fluorescein 5' isothiocyanate. PMID- 3024657 TI - Production of superoxide anion radicals during the oxidative metabolism of amino chloramphenicol. PMID- 3024658 TI - Comparative effects of ethanol and other depressant drugs on membrane order in rat synaptosomes using ESR spectroscopy. AB - The effects of ethanol, t-butanol and pentobarbitone on the membrane order of rat synaptosomal membranes have been compared using 3 spin-label probes, 5-doxyl stearic acid which reports from a lipid site near the membrane surface, 16-doxyl stearic acid which reports from a deeper lipid site, and maleimide-TEMPO which covalently binds to membrane protein. The sensitivity of the membrane proteins to a fluidizing effect of ethanol was increased by lowering the concentration of protein-binding probe. Significant decreases in membrane order were observed at anaesthetic concentrations of ethanol and t-butanol with all three probes; pentobarbitone produced a similar effect but only at very high concentrations. Pentobarbitone caused a marked change in high-field peak shape of the 16-doxyl stearic acid spectra at anaesthetic concentrations; this effect was seen slightly with t-butanol and trichlorethanol but not with ethanol. These studies indicate that the membrane sites of action of ethanol and pentobarbitone as shown by ESR probes are different. PMID- 3024659 TI - GABA enhancement of flunitrazepam binding in mice selectively bred for differential sensitivity to ethanol. AB - The binding of the benzodiazepine [3H]flunitrazepam (FNZ) and the allosteric enhancement of FNZ binding by gamma-aminobutyric acid (GABA) were investigated in brain tissue from short-sleep (SS) and long-sleep (LS) mice, lines selectively bred for differential sensitivity to ethanol. GABA enhanced FNZ binding in a dose dependent manner in both lines. This enhancement was greater in SS than in LS cortical and cerebellar regions, but did not differ between lines in midbrain or hindbrain regions. In whole brain, no difference was observed between the two lines in the number or affinity of benzodiazepine receptors, as determined by FNZ binding. These results suggest that the nature of allosteric interactions within the GABA-benzodiazepine receptor complex is different for LS and SS mice. PMID- 3024660 TI - [Hospital infection caused by rotaviruses in infants, Santiago, Chile]. PMID- 3024661 TI - Approach and escape responses to mesencephalic central gray stimulation in rats: effects of morphine and naloxone. AB - Two experiments were conducted to examine whether regional differences in mesencephalic central gray (CG) could be delineated according to the reinforcing (rewarding and/or punishing) effects of electrical stimulation of CG, and whether such reinforcing properties of CG stimulation would be mediated by endogenous opioid system. In Expt. I, rats with electrodes in CG were trained to turn CG stimulation on and off by running from one compartment to another in a shuttlebox. Rats whose electrode tips were verified at the ventral part of CG, showed both approach and escape responses to the brain stimulation, while those stimulated to more dorsal CG yielded only escape response. In Expt. II, effects of morphine (4 and 8 mg/kg) and naloxone (2 and 10 mg/kg) were investigated for 3 h after injection in the rats which showed both approach and escape responses to CG stimulation in Expt. I. Naloxone significantly increased approach latency to CG stimulation without any effect on escape latency. On the other hand, morphine markedly enhanced the number of shuttling responses and tended to decrease approach latency to stimulation in the late postinjection period, although it increased both approach and escape latencies in the early period. The results were discussed in terms of rewarding and punishing effects of CG stimulation and their underlying endogenous opioid-opiate receptor mechanism. PMID- 3024662 TI - Effects of flunitrazepam on passive avoidance behaviour in mice subjected to immobilization stress or familiarized with the testing apparatus. AB - The effects of flunitrazepam on passive avoidance behaviour were investigated in DBA/2 mice. In a first set of experiments retention performance impairment was observed in mice injected with the drug immediately but not 120 min after training. In a second set of experiments, immobilization stress enhanced, while familiarization with the apparatus decreased, the effects of flunitrazepam, suggesting involvement of emotional factors. All the effects observed were antagonized by naltrexone, showing involvement of opioid receptors. PMID- 3024663 TI - An examination of parkinsonian versus anhedonia contributions to self-stimulation impairments induced by dopamine dysfunction. AB - Interference with brain dopamine neurotransmission can severely impair brain stimulation reward behavior. The significance of this impairment in reward behavior, however, has been a problematic issue. That is, it has been difficult to determine whether the dopamine dysfunction has attenuated the reward effect of the stimulation or has merely rendered the animal less able to generate the behavior required to obtain reinforcement. To experimentally re-assess this issue, the present studies examine the effect of unilateral 6-hydroxydopamine lesions of forebrain dopamine neurons on brain stimulation reward in animals. In general, these studies highlight the substantial motoric deficits produced by the lesion treatments. By combining lesion and neuroleptic drug treatments, however, it appeared that effects on reward could be detected. PMID- 3024664 TI - Modulation of the brain aversive system by GABAergic and serotonergic mechanisms. AB - Experiments performed in our laboratory, using electrical stimulation combined with microinjection of drugs in the dorsal midbrain central grey (CG) of the rat, evidenced that direct stimulation of GABA receptors with locally administered gamma-aminobutyric acid (GABA) or the GABAA receptor agonists 4,5,6,7 tetrahydroisoxazolo[5,4-c]pyridin-3-ol, isoguvacine and muscimol raised the aversive threshold, defined as the lowest electrical current intensity inducing flight or escape behaviour when applied to the dorsal CG. The GABAB receptor agonist baclofen was ineffective. Also, enhancement of endogenous GABA action through local injection of the benzodiazepines chlordiazepoxide and midazolam or of pentobarbital resulted in anti-aversive effects. Ro 15-1788 antagonized both chlordiazepoxide and midazolam, suggesting benzodiazepine receptor mediation. In contrast to pro-GABAergic drugs, microinjection of the GABA antagonists bicuculline and picrotoxin into the CG elicited flight behaviour, like the electrical stimulation. Similar experiments with drugs influencing serotonergic neurotransmission evidenced that intra-CG microinjection of serotonin (5-HT) or of the direct 5-HT receptor agonist 5-methoxy-N,N-dimethyltryptamine increased the aversive threshold. The anti-aversive effect of 5-HT was potentiated by the selective inhibitor of 5-HT neuronal uptake, zimelidine. Also, the latter drug increased the aversive threshold when given alone. The anti-aversive effect of 5 HT was antagonized by local pretreatment with either metergoline or ketanserin, the latter being a selective blocker of 5-HT2 receptors. In contrast to the GABA antagonists mentioned above, the 5-HT receptor blockers did not evoke aversive behaviour per se. Therefore, both GABAergic and serotonergic mechanisms are likely to play an inhibitory role in the dorsal CG integrating aversive behaviour. The former seem to act tonically, whereas 5-HT would act in a phasic way. The implications of these results for the pathophysiology and drug treatment of chronic anxiety, panic states and pain disorders are briefly discussed. PMID- 3024665 TI - Isolation, expression and characterization of a human apolipoprotein B 100 specific cDNA clone. AB - The isolation and characterization of a human apolipoprotein B 100-specific cDNA clone (lambda gt-B1) containing a 1321 base pairs (bp) spanning insert is described. It encodes the 3'-nontranslated 281 bp long region up to the polyadenylation site and 1040 bp of the C-terminal coding region of 345 amino acid residues of human apo B 100 and the stop codon. The lambda gt-B1 cDNA clone has been isolated from a human hepatoma cDNA expression library by immunoscreening using affinity-purified polyclonal anti apo B 100 antibodies. The nucleotide sequence of the apo B 100 insert has been determined. A part of the polypeptide sequence derived from this nucleotide sequence was identical with the amino-acid sequence obtained by protein sequencing of a purified cyanogen bromide fragment of apo B 100. The fusion protein consisting of beta-galactosidase and the 345 amino-acid residue long C-terminus of apo B 100 had an apparent molecular mass of 148 kDa in NaDodSO4 polyacrylamide gel electrophoresis. In Northern blot hybridization analysis the insert of the apo B 100-cDNA clone hybridized to a 20 to 22 kb mRNA from adult human liver. PMID- 3024666 TI - Immunochemical characterization of two activator proteins stimulating enzymic sphingomyelin degradation in vitro. Absence of one of them in a human Gaucher disease variant. AB - Two nonenzymic activator proteins shown previously to strongly stimulate enzymic sphingomyelin degradation in vitro were purified from human Gaucher type 1 and control spleen. Activator A1 (molecular mass 6,500 Da) had affinity for ConA Sepharose, while activator A2 (molecular mass 3,500 Da) did not. Monospecific antibodies to each activator protein were prepared in rabbits by immunization with protein purified from type 1 Gaucher spleen. A1 and A2 activators from Gaucher type 1 spleen were shown to be immunochemically identical to A1 and A2 activators from control spleen. However, A1 and A2 activators, whether isolated from Gaucher type 1 or control spleen, were shown to be distinct proteins. Immunochemical examination of all collected fractions during the purification revealed the existence of a third activator (molecular mass 6,000 Da), which was antigenically identical to A1 activator but had no affinity for ConA-Sepharose. The two forms of A1 activator showed similar mobility on immunoelectrophoresis differing from that of A2 activator. Fibroblast extracts from controls and patients with different variants of Gaucher disease were investigated using immunodiffusion against antisera to A1 or A2 activator. In contrast to normal and Gaucher (types 1, 2 and 3) cell extracts, those of a Gaucher patient with normal glucosylceramidase activity had no visible precipitin line towards the antiserum against the two forms of A1 activator. The lack of crossreacting material to antibodies against A1 activator was confirmed by radial immunodiffusion and rocket immunoelectrophoresis. A1 activator stimulated the basal glucosylceramidase activity 5-6 fold in fibroblasts from this patient, whereas the normal effect was only a 1.2-1.5-fold stimulation. The immunological results together with the biochemical data provide evidence for the lack of an activator protein in a variant form of human Gaucher disease for the first time. PMID- 3024667 TI - Purification and properties of an acetate kinase from Rhodopseudomonas palustris. AB - An acetate kinase from the photolithoautotrophically grown purple bacterium Rhodopseudomonas palustris was purified to apparent homogeneity by use of high resolving liquid chromatography steps. The monomeric enzyme was characterized by a relative molecular mass of 46,500 and an isoelectric point of 4.9. There was an absolute requirement for divalent metal ions in the enzymatic reaction. Mg2+ and Mn2+ were the most activating cations. The acetate kinase used pyrimidine and purine nucleotides almost equally well as phosphoryl donors. The enzyme phosphorylated acetate, propionate, butyrate and isobutyrate. ATP and acetate revealed the lowest apparent Km values and seemed to act as the favoured substrates. The apparent Km values for ATP formation were considerable lower than those for the formation of acetyl phosphate. The activation energy Ea = 21 kJ/mol of the acetyl phosphate formation was determined by application of Arrhenius plots. PMID- 3024668 TI - Inhibition by FOY of kininase II in human plasma. AB - In this investigation we studied the inhibitory effect of FOY S 983 (gabexate mesilate) and FOY S 980 (camostate mesilate) on the kininase II activity in human plasma in vitro. Both compounds were able to inhibit kinase II, however, compared to captopril or EDTA only very weakly. The inhibitory effect of FOY S 983 or FOY S 980 could be diminished neither by NaOH-induced hydrolysis of the inhibitor nor by dialysis or ultrafiltration, but could be clearly reduced by dialysis against ZnCl2 solution. The inhibition of kininase II activity in human plasma by FOY S 983 was due to its 6-guanidinocaproate component probably acting as Zn2+ complexing agent. The ethyl 4-hydroxybenzoate component of FOY S 983 had no inhibitory effect. PMID- 3024669 TI - The galactose-recognizing system of rat peritoneal macrophages. Receptor-mediated binding and uptake of glycoproteins. AB - Binding and phagocytosis of sialidase-treated cells by peritoneal macrophages is mediated by a galactose-specific receptor. So far, only cells or particles exposing terminal galactose residues were demonstrated to be ligands. We present results obtained with a newly developed radio-receptor assay, which proves both binding and uptake of glycoproteins mediated by the galactose-recognizing receptor of peritoneal macrophages. Requirement of Ca2+ for binding is used to distinguish between reversibly surface-bound and irreversibly internalized ligands. By using this approach, the uptake of the ligand is followed and its inhibition with phenylglyoxal and N-ethylmaleimide is demonstrated. Evidence was also obtained that internalization is followed by degradation of the ligand. Studies on the specificity show that only galactose is recognized but that the binding strength depends on the arrangement of galactose residues presented by the ligand. PMID- 3024670 TI - Choosing a treatment modality for the infant, child and adolescent with endstage renal disease. AB - The factors involved in choosing a treatment modality for the infant, child and adolescent with endstage renal disease (ESRD) are different than those utilized when counseling an adult patient. Age at the time ESRD develops, mental status, psychosocial status and the primary renal disease must be taken into consideration when contemplating the optimal therapeutic modality for the pediatric patient with ESRD. PMID- 3024671 TI - [Evaluation of NAD glycohydrolase in leukocytes from peripheral blood and broncho alveolar lavage in man]. PMID- 3024673 TI - Effect of different cell detachment techniques on fibroblast chemotaxis to fibronectin and conditioned medium. PMID- 3024672 TI - [Production of oxygen radicals and the UV spectrum of furanocoumarins]. PMID- 3024674 TI - [Effects of calcium pyrophosphate and hydroxyapatite crystals on the production of oxygen free radicals by polymorphonuclear neutrophil leukocytes]. PMID- 3024675 TI - [Effect of forskolin on the active transport of sodium and on the permeability of skin isolated from Bufo bufo]. PMID- 3024676 TI - Natural killer cells and cytotoxic T lymphocytes activities can be differentiated by their different sensitivities to pituitary hormones in vitro (LH, FSH, ACTH, GH). PMID- 3024677 TI - Vitamin E action on D3 hypervitaminosis. PMID- 3024678 TI - Retinoic acid action on D3 hypervitaminosis. PMID- 3024679 TI - Control of heme internalization and metabolism in tumor cells: evidence for a heme-receptor? PMID- 3024681 TI - [Molecular genetics of Herpes simplex viruses: mapping of the mutation conferring resistance to benzhydrazone and of three syncytial loci simultaneously present in a mutant of the Herpes simplex virus type 1]. PMID- 3024680 TI - [Molecular genetics of the Herpes simplex viruses: physical and functional mapping of a new syncytial locus in the region coding thymidine kinase and glycoprotein H]. PMID- 3024682 TI - [Radioimmunoassay determination of LH, FSH, prolactin, TSH and ACTH in the preovulatory phase under basal conditions and after treatment with a prostaglandin analog]. PMID- 3024683 TI - [Functional characteristics of rat hepatocytes in primary culture]. PMID- 3024684 TI - Oncogenous osteomalacia: a new case secondary to a malignant tumor. AB - A case of oncogenous osteomalacia secondary to a fibrous malignant histiocytoma in a 31-year-old male is described. The patient also demonstrated a lupuslike anticoagulant. Clinical signs of osteomalacia preceded by 9 years those of the tumor, a feature occurring in only 8% of these malignancies. Surgical resection of the tumor and surrounding tissues was followed by a clinical improvement and a return to normal of serum phosphorus and tubular reabsorption of phosphate, though the lupuslike anticoagulant persisted. This first description of a fibrous malignant histiocytoma with associated osteomalacia and lupuslike anticoagulant makes compulsory the inclusion of these syndromes among those already described that may appear with this tumor. PMID- 3024686 TI - Soluble pyrophosphates and remineralisation. PMID- 3024687 TI - Reversible renal failure after combined treatment with enalapril and frusemide in a patient with congestive heart failure. PMID- 3024685 TI - Different metabolism of vitamin D2/D3 in epileptic patients treated with phenobarbitone/phenytoin. AB - Serum concentrations of vitamin D metabolites were measured before and during treatment with either vitamin D2 or vitamin D3, 4000 IU per day for 24 weeks, in 22 epileptic outpatients receiving phenobarbitone/phenytoin. The serum concentration of total 1,25(OH)2D did not change during the treatment period in any of the treatment groups. On the other hand, in the vitamin D2 group, serum 25(OH)D2, total 25(OH)D, and 24,25(OH)2D increased significantly during the trial, whereas serum concentrations of the vitamin D3 metabolites were unchanged. In the vitamin D3 group, serum concentrations of the vitamin D3 metabolites increased significantly, whereas the vitamin D3 metabolite levels remained unchanged. However, vitamin D3 treatment resulted in a 2-4-fold greater increase in serum concentrations compared to vitamin D2 treatment. Treatment with vitamin D2 and vitamin D3 in the same dose in IU results in considerably different serum concentrations of the vitamin D metabolites. PMID- 3024688 TI - Assessment of the Datex Relaxograph during anaesthesia and atracurium-induced neuromuscular blockade. AB - The Datex Relaxograph is a recently introduced electromyographic monitor of neuromuscular transmission. It has been assessed in patients requiring neuromuscular blockade and the results compared with those obtained simultaneously with a force transducer. There was a good correlation between the two methods of measurement for both twitch height and train-of-four ratio (correlation coefficients 0.93 and 0.97, respectively). The Datex Relaxograph proved easy to use in the clinical setting. PMID- 3024689 TI - Effects of a mu-opioid receptor agonist (codeine phosphate) on visuo-motor coordination and dynamic visual acuity in man. AB - Effects of codeine (30, 60 and 90 mg) on visuo-motor coordination and dynamic visual acuity, together with critical flicker fusion, digit symbol substitution, complex reaction time and subjective assessments of mood, were studied from 0.75 2.0 h after ingestion by six healthy female adults. The study was double-blind and placebo controlled, and triprolidine (10 mg) was used as the active control. The effect on visuo-motor coordination was limited and was dose related and linear, and performance was altered on visuo-motor coordination with 60 and 90 mg codeine, and on dynamic visual acuity with 90 mg codeine (P less than 0.05). No other effect of codeine was detected. Modulated neuromuscular function is likely to be the common denominator of the changes in performance with codeine, though nausea, but not sedation, may be a contributory factor. It is possible that altered performance with codeine may involve interactions with different receptors than those which lead to sedation. PMID- 3024690 TI - Plasma angiotensin-converting enzyme activity in patients with bronchial carcinoma. AB - Plasma angiotensin-converting enzyme (ACE) activities were measured in 58 consecutive patients presenting with bronchial carcinoma. The mean ACE activity before treatment was significantly lower than that of a control population (P less than 0.005). There was a significant and direct relationship between the initial plasma ACE activity and survival time (P less than 0.01) which could not be explained by further analysis for age, clinical staging, or respiratory function, as judged by % FEV. There was a significant increase in plasma ACE activity (P less than 0.03) in nine patients with three or more plasma samples after treatment with chemotherapy or radiotherapy. These results suggest that low plasma ACE activity is associated with poor prognosis in bronchial carcinoma. PMID- 3024691 TI - Disseminated intravascular coagulation (DIC) induced by liquoid (polyanetholsulfonate) in the rat. V. Effects on circulating fibronectin. AB - Levels of plasma fibronectin (Fn) were 63% lower than normal 15 min after the intravenous injection of liquoid (P less than 0.01); 3 h later they were still low but rebounded to 35% above normal (P less than 0.01) by 24 h. Concurrently microthrombi containing fibrinogen, Fn and Factor VIII related-antigens (VIII:Ag) were detected in the kidneys and lungs by immunohistopathological studies. Ultrastructurally, thrombi were composed of dense granular and occasional fibrillar non-striated material. In liquoid-injected rats 125I-fibrinogen mainly localized in kidneys and lungs, especially in the latter (P less than 0.01), and the lungs had a higher wet-to-dry weight ratios than did controls (P less than 0.01). It is concluded that the polyanion (liquoid)-induced intravascular coagulation-like reaction sequestered Fn concomitantly with the precipitation of fibrinogen and VII:Ag in the microclots. The reduced concentration of plasma Fn may have impaired the disposal of coagulation products thus enhancing the expression of the coagulopathy-mediated renal and pulmonary histopathology. It is suggested that the liquoid-related coagulopathy may have resulted in enzymatic lysis of Fn. PMID- 3024692 TI - In vitro inhibition of lipopolysaccharide-induced bone resorption by polymyxin B. AB - Lipopolysaccharide (LPS) purified from Haemophilus actinomycetemcomitans stimulates resorption and inhibits collagen synthesis in mouse calvaria bones in vitro. Addition of polymyxin B caused a dose-related inhibition of LPS-stimulated bone resorption and reversal of the inhibition of collagen synthesis. A polymyxin B to LPS ratio of 2:1 prevented bone resorption and restored collagen synthesis to control levels. The activity of polymyxin B was specific for LPS as bone resorption induced by prostaglandin E2 or parathyroid hormone was unaffected at similar concentrations. These results indicate that polymyxin B and its analogues may have potential in the treatment of periodontal disease by reducing bone loss induced by lipopolysaccharides. PMID- 3024693 TI - Myocarditis in mice infected with Coxsackie virus B3. AB - Perimyocarditis in the heart of BALB/c mice infected with Coxsackie virus group B type 3 (CB3) was studied to determine whether it is limited to the right perimyocardium and to show whether or not perimyocarditis or myocardial lesions are produced in both left and right ventricles. CB3 was recovered from the heart on days from 2 to 13 after inoculation, but thereafter no virus was isolated from any part of the heart. Histopathologically, from days 1 to 4, hyaline or granular degeneration and necrosis of the muscle fibres with or without calcium deposits and an inflammatory mononuclear cell infiltration was limited to the right perimyocardium. On days 6 to 18, however, degeneration and necrosis of the muscle fibres and an inflammatory mononuclear cell infiltration were found not only in the right perimyocardium, but also in both left and right ventricular wall, the left perimyocardium, both right and left endomyocardium and the septum. In the right ventricular lesions, the incidence and intensity of the histopathological changes in the perimyocardium were greater than those in the muscular layer or septum. In contrast, in left ventricular lesions, the incidence and intensity of the histopathological changes in the muscular layers were greater than those in the peri- and endo--cardium. It is inferred, therefore, that the right perimyocardium and left ventricular wall are more susceptible to CB3 infection than right ventricular wall or left peri- and endocardium. It is concluded that CB3 can produce not only right-sided perimyocarditis, but also both right and left ventricular lesions and endocardial or septal changes in the mouse heart. PMID- 3024694 TI - Adriamycin and altered membrane functions in rat hearts. AB - To evaluate the potential roles of alterations of membrane functions and resulting calcium overload in the pathogenesis of acute adriamycin cardiotoxicity, we observed sarcolemmal, sarcoplasmic reticular and mitochondrial functions in isolated hearts perfused by the Langendorff technique and exposed to adriamycin. Myocardial tissue calcium content was increased to 120 and 130% of the control level after 30 and 60 min perfusion with adriamycin (50 micrograms/ml), respectively. Sarcolemmal ouabain-sensitive Na+,K+-ATPase activity was decreased by 46% after 30 min perfusion, compared with the control. Mitochondrial calcium uptake was also depressed but sarcoplasmic reticular calcium uptake and binding remained unaltered. Mitochondrial respiratory activity was depressed after 60 min perfusion, when glutamate was used as a substrate, thereby indicating that adriamycin had an inhibitory effect on the NADH dehydrogenase system. Thus, among membrane functions directly or indirectly regulating calcium movement, the inhibition of enzyme systems susceptible to lipid peroxidation may play important roles in calcium overload induced by adriamycin. PMID- 3024695 TI - Follow up study of workers manufacturing chrysotile asbestos cement products. AB - A cohort study has been carried out of 2167 subjects employed between 1941 and 1983 at an asbestos cement factory in England. The production process incorporated the use of chrysotile asbestos fibre only, except for a small amount of amosite during four months in 1976. Measured airborne fibre concentrations available since 1970 from personal samplers showed mean levels below 1 fibre/ml, although higher levels had probably occurred previously in certain areas of the factory. No excess of lung cancer was observed in the mortality follow up by comparison with either national or local death rates, and analyses of subgroups of the workforce by job, exposure level, duration of employment, duration since entry, or calendar years of employment gave no real suggestion of an asbestos related excess for this cause of death. There was one death from pleural mesothelioma and one with asbestosis mentioned as an associated cause on the death certificate, but neither is thought to be linked to asbestos exposure at this factory. Other suggested asbestos related cancers, such as laryngeal and gastrointestinal, did not show raised risks. Although the durations of exposure were short in this study, the findings are consistent with two other studies of workers exposed to low concentrations of chrysotile fibre in the manufacture of asbestos cement products which reported no excess mortality. PMID- 3024696 TI - Characterization of gonadotrophin receptors in a testosterone-producing ovarian androblastoma. Case report. PMID- 3024697 TI - A recent epidemic of Coxsackie virus type A24 acute haemorrhagic conjunctivitis in Singapore. AB - A recent epidemic of acute conjunctivitis in Singapore showed again the importance of Coxsackie virus type A24 variant as a causative agent of acute haemorrhagic conjunctivitis (AHC). Although the ocular manifestations appeared similar to those described for the 1970 and 1975 outbreaks, a markedly higher rate of respiratory involvements was noted. Not observed in previous epidemics were herpes-like vesicles in the conjunctiva and eyelids of one patient and vesicles in the buccal mucosa and lips of another from whom Coxsackie virus A24 was isolated. The most interesting finding in this study was the isolation of five wild (non-Sabin) poliovirus type 1 strains. Three strains were obtained from conjunctival and two from throat swabs of patients with mild to severe conjunctivitis. It is conceivable that the rare reports of polio-like paralysis or radiculomyelitis accompanying or following AHC in a few Asian countries could be attributed to concurrent infections with a poliovirus and either enterovirus type 70 or Coxsackie virus type A24. PMID- 3024698 TI - Gardner's dento-maxillary stigmas in patients with familial adenomatosis coli. AB - Fifty patients with familial adenomatosis coli (FAC) studied by panoramic tomography (PTG) showed osteomatous jaw changes in 82% as compared with 10% in matched controls. Supernumerary teeth, compound odontomas and/or impacted teeth were seen in 30% of the patients compared with 4% of the controls. A correlation was found between dental abnormalities and the large number of osteomas. The results support the view that both osteomas of the jaws and dental abnormalities are features of FAC. PTG of the jaws may serve as a valuable tool for early detection of FAC by oral surgeons, at least in the case of the most striking changes (about 20%). PMID- 3024699 TI - Protein disulfide-isomerase retains procollagen prolyl 4-hydroxylase structure in its native conformation. AB - Protein disulfide-isomerase was isolated as a homogeneous protein from 15-day-old chick embryos. The enzyme has a molecular weight of 56,000 in SDS-polyacrylamide gel electrophoresis. Its Km value for randomly cross-linked ribonuclease, a protein used as a substrate for the enzyme, was 0.3 microM, and the Km value for DTT was 1.0 microM. Its optimum pH was 7.5 and its optimum temperature, 33 degrees C. The maximal velocity of pure protein disulfide-isomerase from chick embryos under optimal conditions was about 29,000 units/g. Protein disulfide isomerase was able to activate purified prolyl 4-hydroxylase 2- to 3-fold, the activation being higher for enzyme stored for a longer time. This activation is probably due to the repairing of disulfide exchanges occurring in the prolyl 4 hydroxylase structure during purification and storage. Prolyl 4-hydroxylase activity was very stable in microsomes, however, and protein disulfide-isomerase was unable to increase the microsomal prolyl 4-hydroxylase activity, suggesting that prolyl 4-hydroxylase retains its native conformation in microsomes. Protein disulfide-isomerase was able to reactivate prolyl 4-hydroxylase inactivated by mild H2O2 treatment. The activity obtained after this treatment and protein disulfide-isomerase incubation corresponded to the amount of prolyl 4-hydroxylase tetramer found after H2O2 treatment. The data suggest that protein disulfide isomerase is able to activate only the tetramer part of the enzyme preparation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3024700 TI - Phosphate inhibition of the copper- and zinc-containing superoxide dismutase: a reexamination. AB - Phosphate was reported to be an inhibitor of copper- and zinc-containing superoxide dismutase (SOD) [de Freitas, D.M., & Valentine, J.S. (1984) Biochemistry 23, 2079-2082]. Thus SOD activity, in 50 mM 4-(2-hydroxyethyl)-1 piperazineethanesulfonic acid (HEPES) (pH 7.4), was decreased by approximately 50% when the assay was made 10 mM in phosphate, and the ionic strength was adjusted with sodium fluoride. The inhibitory effect of phosphate was attributed to the neutralization of the positive charge on the guanidino residue of Arg-141. We have reexamined the effects of phosphate inhibition of SOD and found that the enzyme has identical activity in phosphate or HEPES buffer when the ionic strength is adjusted with NaBr. The putative inhibitory effect of phosphate appears to have been due to fluoride inhibition of the superoxide generating system of xanthine/xanthine oxidase. We have confirmed this result by using a photochemical generation of O2- in addition to the enzymatic generation of O2-. Chemical modification of the lysine residues to homoarginines does not affect the activity of the enzyme and does not impart a phosphate sensitivity. Chemical modification with phenylglyoxal caused approximately 80% inactivation of the native enzyme and 90% inactivation of the O-methylisourea-modified enzyme. Our results suggest that phosphate does not inhibit the copper- and zinc-containing superoxide dismutase (Cu,Zn-SOD) beyond the expectations of its effect on ionic strength. PMID- 3024701 TI - Nonionic detergents increase the stoichiometry of ligand binding to the rat hepatic galactosyl receptor. AB - When digitonin is used to expose intracellular galactosyl (Gal) receptors in isolated rat hepatocytes, only about half of the binding activity for 125I asialoorosomucoid (ASOR) is found as compared to cells solubilized with Triton X 100. The increased ligand binding in the presence of detergent is not due to a decrease in Kd but could be due either to an increase in the number of ASORs bound per receptor or to exposure of additional receptors. Several experiments support the former explanation. No additional activity is exposed even when 80% of the total cell protein is solubilized with 0.4% digitonin. It is, therefore, unlikely that receptors are in intracellular compartments not permeabilized by digitonin and inaccessible to 125I-ASOR. Digitonin-treated cells are not solubilized by Triton X-100 if they are first treated with glutaraldehyde under conditions that retain specific binding activity. 125I-ASOR binding to these permeabilized/fixed cells increases about 2-fold in the presence of Triton X-100 and a variety of other detergents (e.g., Triton X-114, Nonidet P-40, Brij-58, and octyl glucoside) but not with the Tween series, saponin, or other detergents. When these fixed cells are washed to remove detergent, 125I-ASOR binding decreases almost to the initial level. Affinity-purified Gal receptor linked to Sepharose 4B binds approximately twice as much 125I-ASOR in the presence of Triton X-100 as in its absence. The results suggest that the increase in Gal receptor activity in the presence of nonionic detergents is due to an increase in the valency of the receptor rather than to exposure of additional receptors. PMID- 3024702 TI - Nucleotide sequence and nuclease hypersensitivity of the Chinese hamster dihydrofolate reductase gene promoter region. AB - We have sequenced the 1240 base pairs (bp) upstream from the translation start site of the hamster dihydrofolate reductase (DHFR) gene. The DNA in the 5' flanking region contains several elements that are homologous in both sequence and relative location to corresponding elements in the human and murine DHFR genes: an 11-bp element adjacent to the ATG codon, a 19-bp element that coincides with the major transcription start site, and two 29-bp upstream elements that are represented 4 times in the murine DHFR gene but only once in the human gene. Two clusters of short, G/C-rich elements conforming to the consensus binding sequence for the transcription factor Spl are located in the upstream region in all three genes. The symmetrical placement of the G/C boxes coincides with a symmetrical DNase I hypersensitive pattern in the chromatin, suggesting that the Spl protein may be involved in maintaining chromatin structure in this region. PMID- 3024703 TI - Amino acid sequence of a chicken heat shock protein derived from the complementary DNA nucleotide sequence. AB - The complete nucleotide sequence for a chicken heat shock protein (hsp108) was determined from cDNA clones isolated from hen oviduct and bursal lymphoma recombinant DNA libraries. This protein has certain biochemical similarities to the progesterone receptor, but it is clearly distinct from it. The initial cDNA clone, isolated from a chicken oviduct cDNA library, was detected by antibody screening and hybrid-selected translation [Zarucki-Schulz, T., Kulomaa, M. S., Headon, D. R., Weigel, N. L., Baez, M., Edwards, D. P., McGuire, W. L., Schrader, W. T., & O'Malley, B. W. (1984) Proc. Natl. Acad. Sci. U.S.A. 81, 6358-6362]. The earlier clones were used to screen for additional cDNAs, and cDNAs that define the entire mRNA sequence of hsp108 have been obtained. The nucleotide sequence codes for peptides present in hsp108 as determined by protein microsequencing. The 5' end of the mRNA was determined by primer extension studies. The mRNA contains a noncoding region of 101 nucleotides upstream from the predicted initiation codon. The 3' untranslated region contains 244 nucleotides beyond the termination codon, and it contains a predicted polyadenylation signal 26 nucleotides from the end of the complete cDNA. The coding region of 2385 nucleotides corresponds to a polypeptide chain of 795 amino acids, giving a molecular weight of 91,555 for the hsp108 protein. In another paper, evidence is presented that hsp108 shows a high degree of amino acid sequence homology with two heat shock proteins, hsp90 (yeast) and hsp83 (Drosophila), and is indeed inducible by heat shock [Sargan, D. R., Tsai, M.-J., & O'Malley, B. W. (1986) Biochemistry (following paper in this issue)]. PMID- 3024704 TI - Inhibition of vesicular stomatitis virus protein synthesis and infection by sequence-specific oligodeoxyribonucleoside methylphosphonates. AB - Oligodeoxyribonucleoside methylphosphonates which have sequences complementary to the initiation codon regions of N, NS, and G vesicular stomatitis virus (VSV) mRNAs were tested for their ability to inhibit translation of VSV mRNA in a cell free system and in VSV-infected mouse L cells. In a rabbit reticulocyte lysate cell-free system, the oligomers complementary to N (oligomer I) and NS (oligomer II) mRNAs inhibited translation of VSV N and NS mRNAs whereas oligomer III had only a slight inhibitory effect on N protein synthesis. At 100 and 150 microM, oligomer I specifically inhibited N protein synthesis in the lysate. In contrast, at 150 microM, oligomer II inhibited both N and NS protein synthesis. This reduced specificity of inhibition may be due to the formation of partial duplexes between oligomer II and VSV N mRNA. The oligomers had little or no inhibitory effects on the synthesis of globin mRNA in the same lysate system. Oligomers I III specifically inhibited the synthesis of all five viral proteins in VSV infected cells in a concentration-dependent manner. The oligomers had no effects on cellular protein synthesis in uninfected cells nor on cell growth. An oligothymidylate which forms only weak duplexes with poly(rA) had just a slight effect on VSV protein synthesis and yield of virus. Oligomers I-III have extensive partial complementarity with the coding regions of L mRNA. The nonspecific inhibition of viral protein synthesis in infected cells may reflect the role of N, NS, and/or L proteins in the replication and transcription of viral RNA or result from duplex formation between the oligomers and complementary, plus-strand viral RNA.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3024705 TI - External anions regulate stilbene-sensitive proton transport in placental brush border vesicles. AB - The mechanism for HCO3-(-)independent proton permeability in microvillus membrane vesicles (MVV) isolated from human placenta was examined by using the entrapped pH indicator 6-carboxyfluorescein (6CF). Proton fluxes (JH) across MVV were determined in response to induced pH and anion gradients from the time course of 6CF fluorescence, the MVV buffer capacity, and the 6CF vs. pH calibration. In the absence of anions, JH was 12 +/- 2 nequiv s-1 (mg of protein)-1 (pHin 7.4, pHout 6.0, MVV voltage-clamped with K+/valinomycin, 23 degrees C), corresponding to a proton permeability coefficient of 0.02 cm/s, with an activation energy of 9.1 +/ 0.3 kcal/mol. JH was inhibited 20% by dihydro-4,4'-diisothiocyano-2,2' stilbenedisulfonic acid (H2DIDS) with KI = 8 microM [( Cl-]out = 0 mM). For a 0.5 unit pH gradient JH increased from 1.5 to 4.6 nequiv s-1 (mg of protein)-1 as the internal MVV pH was increased (5.5-7.5). External Cl-, Br-, and I- (but not SO4(2 ) and PO4-) increased JH 1.3-2.5-fold for both inwardly and outwardly directed pH gradients with KD = 1.0 +/- 0.4 mM (Br-) and greater than 100 mM (Cl-). This increase was blocked by 100 microM H2DIDS but not by amiloride or furosemide. Internal Cl- did not alter JH induced by pH gradients nor were proton fluxes induced by anion gradients in the absence of a pH gradient. Experiments in which JH was driven by membrane potentials (induced by valinomycin and K+ gradients) indicated that proton transport was voltage-sensitive. These experiments demonstrate a stilbene-sensitive electrogenic proton transport mechanism in MVV that is regulated allosterically by anions at an external binding site. PMID- 3024706 TI - Characterization of a fragment of bovine von Willebrand factor that binds to platelets. AB - Bovine von Willebrand factor was digested with human plasmin in order to isolate and characterize a fragment that can bind to human platelets. A terminal plasmin digest of bovine von Willebrand factor is composed of five fragments, ranging in relative molecular weight (Mr) from 250,000 to 35,000. The major fragment has a Mr of 250,000 and consists of four disulfide-linked polypeptide chains with Mr from 69,000 to 35,000. The Mr 69,000 and 49,000 polypeptides possess carbohydrate moieties, as indicated by their reaction with periodate-Schiff reagent. Gel filtration studies suggest that, at physiological ionic strength, four of the Mr 250,000 fragments associate into a limited noncovalent oligomer. Monoclonal antibodies were prepared against native von Willebrand factor and used to characterize the distribution of epitopes on native vWF and the Mr 250,000 major fragment. Two of the monoclonal antibodies that recognize the major fragment (2 and H-9) inhibit platelet agglutination. The Mr 250,000 fragment binds to human platelets, and the binding is inhibited by monoclonal antibodies 2 and H-9. The Mr 250,000 fragment does not agglutinate platelets, consistent with a requirement for high molecular weight oligomers of von Willebrand factor for platelet agglutination. The Mr 250,000 fragment can compete with intact, bovine von Willebrand factor for binding to human platelets. However, its affinity is one tenth that of intact von Willebrand factor. PMID- 3024707 TI - Molecular mechanism of opioid receptor selection. AB - Preferred conformations, orientations, and accumulations of 26 opioid peptides on lipid membranes were estimated and compared with pharmacologic and selective binding data taken from the literature. Interaction with mu-receptors was governed by the net positive charge effective at the message domain of the agonist peptides z(eff) as the Boltzmann term ez(eff) that determines relative accumulation on anionic biologic membranes. Selection for delta-receptors was reduced by z(eff) and correlated with e-z(eff). Selection for kappa-receptors was governed by the peptide amphiphilic moment A. A pronounced scalar magnitude A and almost perpendicular orientation of the N-terminal message domain as an alpha helix were favorable for kappa-site selection. Potencies as kappa-agonists and binding affinities correlated with A X ez(eff). The classical site selectivity caused by the receptor requirements for a complementary fit of the agonist to the discriminator site is thus crucially supplemented by a selection mechanism based on peptide membrane interactions (membrane requirements). In the model presented here, the delta-site is exposed to the aqueous compartment surrounding the target cell at a distance comparable to or greater than the Debye-Huckel length and is in a cationic vicinity. The mu-site is exposed to the anionic fixed-charge compartment of the membrane in aqueous surroundings. The kappa-site is buried in a more hydrophobic membrane compartment close to the fixed-charge compartment. The relative accumulation of the opioid message domains in these compartments is determined by the address domains and constitutes a major part of the site selection mechanism.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3024708 TI - Half-life of synovial cell collagenase mRNA is modulated by phorbol myristate acetate but not by all-trans-retinoic acid or dexamethasone. AB - As part of our studies on the mechanisms controlling the synthesis of the neutral proteinase collagenase by rabbit synovial cells, we used a cDNA clone to measure total collagenase mRNA levels and to determine mRNA half-life. Phorbol myristate acetate was used to induce collagenase synthesis while all-trans-retinoic acid and dexamethasone were used to inhibit it. Cells stimulated with phorbol myristate acetate contained substantial amounts of collagenase mRNA, but cells treated with all-trans-retinoic acid or dexamethasone contained decreased amounts of collagenase mRNA which correlated well with levels of collagenase protein. Studies on mRNA half-life showed that the t1/2 for total poly(A+) RNA was about 25 h, while that of collagenase varied from as short as 12 h to as long as 36 h. The half-life was not affected by treatment with all-trans-retinoic acid or dexamethasone but was affected by the level of induction of collagenase mRNA: the greater the amount of collagenase mRNA induced, the longer the t1/2. We conclude that our data are consistent with the hypothesis that retinoic acid and dexamethasone act at the level of transcription to decrease collagenase production and the increased level of collagenase mRNA resulting from stimulation with phorbol esters is, in part, due to increased stability of the induced collagenase mRNA. PMID- 3024709 TI - Binding of amines to the O2-evolving center of photosystem II. AB - The binding of several primary amines to the O2-evolving center (OEC) of photosystem II (PSII) has been studied by using low-temperature electron paramagnetic resonance (EPR) spectroscopy of the S2 state. Spinach PSII membranes treated with NH4Cl at pH 7.5 produce a novel S2-state multiline EPR spectrum with a 67.5-G hyperfine line spacing when the S2 state is produced by illumination at 0 degrees C [Beck, W. F., de Paula, J. C., & Brudvig, G. W. (1986) J. Am. Chem. Soc. 108, 4018-4022]. The altered hyperfine line spacing and temperature dependence of the S2-state multiline EPR signal observed in the presence of NH4Cl are direct spectroscopic evidence for coordination of one or more NH3 molecules to the Mn site in the OEC. In contrast, the hyperfine line pattern and temperature dependence of the S2-state multiline EPR spectrum in the presence of tris(hydroxymethyl)aminomethane, 2-amino-2-ethyl-1,3-propanediol, or CH3NH2 at pH 7.5 were the same as those observed in untreated PSII membranes. We conclude that amines other than NH3 do not readily bind to the Mn site in the S2 state because of steric factors. Further, NH3 binds to an additional site on the OEC, not necessarily located on Mn, and alters the stability of the S2-state g = 4.1 EPR signal species. The effects on the intensities of the g = 4.1 and multiline EPR signals as the NH3 concentration was varied indicate that both EPR signals arise from the same paramagnetic site and that binding of NH3 to the OEC affects an equilibrium between two configurations exhibiting the different EPR signals.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3024710 TI - Effect of the 17- and 23-kilodalton polypeptides, calcium, and chloride on electron transfer in photosystem II. AB - Electron paramagnetic resonance (EPR) measurements were performed on photosystem II (PSII) membranes that were treated with 2 M NaCl to release the 17- and 23 kilodalton (kDa) polypeptides. By using 75 microM 3-(3,4-dichlorophenyl)-1,1 dimethylurea to limit the photosystem II samples to one stable charge separation in the temperature range of 77-273 K, we have quantitated the EPR signals of the several electron donors and acceptors of photosystem II. It was found that removal of the 17- and 23-kDa polypeptides caused low potential cytochrome b559 to become fully oxidized during the course of dark adaptation. Following illumination at 77-130 K, one chlorophyll molecule per reaction center was oxidized. Between 130 and 200 K, both a chlorophyll molecule and the S1 state were photooxidized and, together, accounted for one oxidation per reaction center. Above 200 K, the chlorophyll radical was unstable. Oxidation of the S1 state gave rise to the S2-state multiline EPR signal, which arises from the Mn site of the O2-evolving center. The yield of the S2-state multiline EPR signal in NaCl-washed PSII membranes was as high as 93% of the control, untreated PSII membranes, provided that both Ca2+ and Cl- were bound. Furthermore, the 55Mn nuclear hyperfine structure of the S2-state multiline EPR signal was unaltered upon depletion of the 17- and 23-kDa polypeptides. In NaCl-washed PSII samples where Ca2+ and/or Cl- were removed, however, the intensity of the S2-state multiline EPR signal decreased in parallel with the fraction of PSII lacking bound Ca2+ and Cl-.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3024711 TI - Electron-transfer reactions between flavodoxin semiquinone and c-type cytochromes: comparisons between various flavodoxins. AB - As an extension of previous work from this laboratory using Clostridium pasteurianum flavodoxin [Tollin, G., Cheddar, G., Watkins, J. A., Meyer, T. E., & Cusanovich, M. A. (1984) Biochemistry 23, 6345-6349], we have measured the rate constants as a function of ionic strength for electron transfer from the semiquinones of Clostridium MP, Anacystis nidulans, and Azotobacter vinelandii flavodoxins to the following oxidants: cytochrome c from tuna and horse, Paracoccus denitrificans cytochrome c2, Pseudomonas aeruginosa cytochrome c-551, and ferricyanide. The rate constants extrapolated to infinite ionic strength (k infinity) for the C. MP flavodoxin are all slightly smaller than for the C. pasteurianum flavodoxin, as would be predicted on the basis of the higher redox potential of the C. MP protein. This indicates that there is a close similarity between the surface topographies of the two proteins in the vicinity of the coenzyme binding site. Moreover, the electrostatic interactions between the two flavodoxins and the various oxidants are also approximately the same. These studies justify our previous use of the crystallographic structure of the C. MP flavodoxin to interpret kinetic results obtained with the structurally uncharacterized C. pasteurianum flavodoxin. Despite their lower redox potentials, both Anacystis and Azotobacter flavodoxins are appreciably less reactive toward all of these oxidants (as much as 2 orders of magnitude in some cases) than are the Clostridium flavodoxins.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3024712 TI - Synthesis of in vitro Co1E1 transcripts with 5'-terminal ribonucleotides that exhibit noncomplementarity with the DNA template. AB - A region that forms the S1 nuclease site in Co1E1 DNA is shown to code for an in vitro transcript, called S1 RNA-B, which contains a 5'-terminal GTP residue that exhibits noncomplementarity with the template's DNA sequence. The synthesis of S1 RNA-B initiates four bases upstream from the start point for S1 RNA-C. The initial four bases in S1 RNA-B and S1 RNA-C are identical. The relative synthesis of S1 RNA-B to S1 RNA-C is sensitive to the concentration of GTP, a substrate that is required for elongation past the +4 position in S1 RNA-C. Dinucleotides that are expected to only initiate synthesis of S1 RNA-C yield two transcripts that appear to initiate from the S1 RNA-C and S1 RNA-B start sites. In vitro studies involving other Co1E1 transcripts, RNA-B and RNA-C, provide similar observations concerning the noncomplementary initiation phenomenon. A model involving transcriptional slippage is suggested to explain the noncomplementary initiation phenomenon. The model proposes that the cycling reaction of Escherichia coli RNA polymerase produces tetranucleotides that are transposed to nearby upstream sequences for priming transcription. PMID- 3024713 TI - Site specificity of psoralen-DNA interstrand cross-linking determined by nuclease Bal31 digestion. AB - A novel method for determination of psoralen photo-cross-linking sites in double stranded DNA is described, which is based on a pronounced inhibition of Bal31 exonuclease activity by psoralen-DNA interstrand cross-links. The results using a 51 base pair fragment of plasmid pUC19 and a 346 base pair fragment of pBR322 show that 5'-TA sequences are preferred cross-linking sites compared to 3'-TA sequences. They also indicate that sequences flanking the 5'-TA site influence the cross-linking efficiency at the site. The DNA photo-cross-linking by 4,5',8 trimethylpsoralen and 8-methoxypsoralen was analyzed, and these two psoralens showed identical site specificity. The 5'-TA preference is rationalized on the basis of the local DNA structure in terms of the pi-pi electronic interaction between the thymines and the intercalated psoralens, as well as on the base tilt angles of the DNA. PMID- 3024714 TI - Phenylalanine hydroxylase from Chromobacterium violaceum is a copper-containing monooxygenase. Kinetics of the reductive activation of the enzyme. AB - Pterin-dependent phenylalanine hydroxylase from Chromobacterium violaceum contains a stoichiometric amount of copper (Cu2+, 1 mol/mol of enzyme). Electron paramagnetic resonance spectroscopy of the enzyme indicates that it is a type II copper-containing protein. The oxidized enzyme must be reduced by a single electron to be catalytically active. Dithiothreitol was found to be an effective reducing agent for the enzyme. Electron paramagnetic resonance data and kinetic results indicate the formation of an enzyme-thiol complex during the aerobic reduction of the enzyme by dithiothreitol. 6,7-Dimethyltetrahydropterin also reductively activates the enzyme, but only in the presence of the substrate, and is kinetically less effective than dithiothreitol. The metal center is not reoxidized as a result of normal turnover. However, the data indicate an alternative pathway exists that results in slow reoxidation of the enzyme. The 4a hydrate of 6-methyltetrahydropterin (4a-carbinolamine) is observed during turnover of the enzyme. This intermediate is also observed during the reaction catalyzed by the iron-containing mammalian enzyme, suggesting that the mechanism of oxygen activation is similar for both enzymes. PMID- 3024715 TI - Synthesis and characterization of ubiquitin ethyl ester, a new substrate for ubiquitin carboxyl-terminal hydrolase. AB - A new substrate for ubiquitin carboxyl-terminal hydrolase, the carboxyl-terminal ethyl ester of ubiquitin, has been synthesized by a trypsin-catalyzed transpeptidation. In the presence of 1.6 M glycylglycine ethyl ester, trypsin removes the carboxyl-terminal glycylglycine of ubiquitin and replaces it with the dipeptide ester. The equilibrium mixture under these conditions contains 30% ubiquitin ethyl ester and 70% hydrolysis product, the 74-residue fragment of ubiquitin. Ubiquitin ethyl ester can be purified by gel filtration and ion exchange chromatography. The structure of this product has been verified by identification of the products of base hydrolysis, tryptic cleavage in aqueous solution, and peptide mapping. When ubiquitin ethyl ester is incubated with purified ubiquitin carboxyl-terminal hydrolase, specific cleavage of the ester linkage is observed. A rapid, sensitive assay is described utilizing high performance liquid chromatography. By use of this assay, it has been shown that ubiquitin carboxyl-terminal hydrolase is inactivated in the absence of thiols. Optimal protective effects are seen with 10 mM dithiothreitol. The rate of catalysis is maximal at pH 8.5, with evidence for catalytically important groups with pK values of 5.2, 7.6, and 9.5. These findings are consistent with the participation of a thiol group in the active site. Native ubiquitin is a competitive inhibitor of ubiquitin ethyl ester hydrolysis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3024716 TI - Regulation of rabbit liver fructose-1,6-bisphosphatase by metals, nucleotides, and fructose 2,6-bisphosphate as determined from fluorescence studies. AB - The fluorescent nucleotide analogue formycin 5'-monophosphate (FMP) inhibits rabbit liver fructose-1,6-bisphosphatase (I50 = 17 microM, Hill coefficient = 1.2), as does the natural regulator AMP (I50 = 13 microM, Hill coefficient = 2.3), but exhibits little or no cooperativity of inhibition. Binding of FMP to fructose-1,6-bisphosphatase can be monitored by the increased fluorescence emission intensity (a 2.7-fold enhancement) or the increased fluorescence polarization of the probe. A single dissociation constant for FMP binding of 6.6 microM (4 sites per tetramer) was determined by monitoring fluorescence intensity. AMP displaces FMP from the enzyme as evidenced by a decrease in FMP fluorescence and polarization. The substrates, fructose 6-phosphate and fructose 1,6-bisphosphate, and inhibitors, methyl alpha-D-fructofuranoside 1,6 bisphosphate and fructose 2,6-bisphosphate, all increase the maximal fluorescence of enzyme-bound FMP but have little or no effect on FMP binding. Weak metal binding sites on rabbit liver fructose-1,6-bisphosphatase have been detected by the effect of Zn2+, Mn2+, and Mg2+ in displacing FMP from the enzyme. This is observed as a decrease in FMP fluorescence intensity and polarization in the presence of enzyme as a function of divalent cation concentration. The order of binding by divalent cations is Zn2+ = Mn2+ greater than Mg2+, and the Kd for Mn2+ displacement of FMP is 91 microM. Methyl alpha-D-fructofuranoside 1,6 bisphosphate, as well as fructose 6-phosphate and inorganic phosphate, enhances metal-mediated FMP displacement from rabbit liver fructose-1,6 bisphosphatase.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3024717 TI - Temperature jump as a new technique to study the kinetics of fast transport of protons across membranes. AB - Application of a temperature jump (2.5 degrees C) to a suspension of liposomes, having phosphate (delta pK/delta T approximately 0.005) as the internal buffer and tris(hydroxymethyl)aminomethane (delta pK/delta T approximately 0.031) as the external buffer, created a delta pH (pHin - pHout) of positive sign in ca. 5 microseconds. Decay of this delta pH was monitored by using the fluorescent pH indicator 8-hydroxy-1,3,6-pyrenetrisulfonic acid entrapped inside the liposome. This technique is useful to study transmembrane proton movement in the time range 5 microseconds-10 s at physiological pH values. The kinetics of proton transport aided by ion carriers such as nigericin, monensin, carbonyl cyanide m chlorophenylhydrazone (CCCP), and valinomycin were studied by our method. The electrogenic nature of transport by CCCP and valinomycin and electroneutral ion transport by nigericin and monensin were shown. From the kinetics of proton transport aided by gramicidin, the time-averaged single-channel conductance of gramicidin channels was estimated to be (2.1 +/- 0.5) X 10(-16) S for H+ at pH 7.5. PMID- 3024718 TI - Immunochemical detection of guanine nucleotide binding proteins mono-ADP ribosylated by bacterial toxins. AB - Rabbits immunized with ADP-ribose chemically conjugated to carrier proteins developed antibodies reactive against guanine nucleotide binding proteins (G proteins) that had been mono-ADP-ribosylated by bacterial toxins. Antibody reactivity on immunoblots was strictly dependent on incubation of substrate proteins with both toxin and NAD and was quantitatively related to the extent of ADP-ribosylation. Gi, Go, and transducin (ADP-ribosylated by pertussis toxin) and elongation factor II (EF-II) (ADP-ribosylated by pseudomonas exotoxin) all reacted with ADP-ribose antibodies. ADP-ribose antibodies detected the ADP ribosylation of an approximately 40-kilodalton (kDa) membrane protein related to Gi in intact human neutrophils incubated with pertussis toxin and the ADP ribosylation of an approximately 90-kDa cytosolic protein, presumably EF-II, in intact HUT-102 cells incubated with pseudomonas exotoxin. ADP-ribose antibodies represent a novel tool for the identification and study of G proteins and other substrates for bacterial toxin ADP-ribosylation. PMID- 3024720 TI - Calorimetric and fatty acid spin label study of subgel and interdigitated gel phases formed by asymmetric phosphatidylcholines. AB - Several saturated asymmetric and symmetric phosphatidylcholines were studied by ESR spectroscopy and differential scanning calorimetry in order to determine the behavior of a fatty acid spin labeled near its terminal methyl, 16-doxylstearate, in the mixed interdigitated gel phase and the Lc subgel phase and other properties of these lipids. This spin label was motionally restricted in the mixed interdigitated gel phases of 18:10PC and 18:12PC. The motional restriction was similar to that reported earlier for fully interdigitated phases. This spin label was motionally restricted almost to the same degree in 10:18PC suggesting that this asymmetric lipid may also form a mixed interdigitated bilayer. In contrast the spin label had more motion in the gel phase of 18:14PC than in symmetric forms of PC, consistent with conclusions from X-ray diffraction studies that this less asymmetric lipid does not form a mixed interdigitated phase. The spin label was partially frozen out of the Lc subgel phases of symmetric forms of PC and 18:14PC formed by storage at low temperature. The phase behavior of the other asymmetric lipids also depended on the sample history. Storage at low temperature caused 10:18PC and 18:12PC to go into ordered phases. The enthalpy of the transition of these ordered phases to the liquid-crystalline phase was 2-2.4 times greater than that of the transition of the gel phase formed on cooling back from the liquid-crystalline phase. The temperature of this high enthalpy transition was 0.8 K below that of the lower enthalpy gel to liquid-crystalline phase transition for 18:12PC, but 4.6 K higher for 10:18PC. The spin label was frozen out of these ordered phases, as it was out of the Lc subgel phases, suggesting that 18:12PC and 10:18PC may also form an Lc phase. 18:10PC was not observed to form an ordered phase although storage of the sample at low temperatures did affect the temperature of its transition from the liquid crystalline phase back to the gel phase upon cycling through its phase transition. PMID- 3024719 TI - Rapid incorporation of the solubilized dihydropyridine receptor into phospholipid vesicles. AB - We describe the rapid incorporation of the CHAPS solubilized dihydropyridine receptor into phospholipid vesicles. A series of sucrose gradient sedimentation experiments demonstrate that the (+)-[3H]PN200-110-labeled dihydropyridine receptor is associated with lipid vesicles following detergent removal by Extracti-gel chromatography. Solubilization of the receptor results in a loss of (+)-[3H]PN200-110 binding affinity relative to that observed in native membranes; the high affinity binding of (+)-[3H]PN200-110 can be restored upon reincorporation of the receptor into phospholipid vesicles. Similarly, the incorporation of the receptor restores its stability to incubation at 37 degrees C relative to that of the detergent solubilized receptor, thereby mimicking the properties of the membrane bound form of the receptor. The dissociation rate of (+)-[3H]PN200-110 from the reconstituted receptor is shown to be allosterically regulated by verapamil and diltiazem, indicating that the binding sites for these calcium antagonists have been inserted along with the dihydropyridine receptor into phospholipid vesicles. The results presented in this report, thus demonstrate the successful reconstitution of the dihydropyridine receptor into phospholipid vesicles by a variety of criteria. The reconstitution method described here is rapid and efficient, and should now facilitate structure function studies of this receptor and its interrelationships with other regulatory components of the voltage-sensitive calcium channel system. PMID- 3024721 TI - Membrane ordering effects of the anticancer agent VM-26. AB - The effect of the anticancer agent VM-26 on acyl chain order of cellular and model membranes was examined by electron spin resonance techniques. The order parameter for the paramagnetic probe 5-doxyl stearate was increased when VM-26 was incorporated into the bilayer of fluid-phase dimyristoylphosphatidylcholine (DMPC) or gel-phase dipalmitoylphosphatidylcholine (DPPC) liposomes at concentrations up to 4.8 mol%. The ordering effect of VM-26 in DMPC was greater than that of cholesterol on an equimolar basis. The less cytotoxic congener of VM 26, VP-16, was only one-third as active as VM-26 in its ordering effects on DMPC. Higher order parameters for 5-doxyl stearate were also noted in asolectin liposomes, Ehrlich ascites tumor cells, and CCRF-CEM cells treated with VM-26. We conclude that VM-26 has significant membrane associated activity in addition to its previously recognized nuclear effects. PMID- 3024722 TI - The role of sphingomyelin synthetase and sphingomyelinase in 1,2 dimethylhydrazine-induced lipid alterations of rat colonic plasma membranes. AB - Recently, our laboratory, utilizing the 1,2-dimethylhydrazine model of colonic adenocarcinoma, demonstrated alterations in the 'dynamic component' of fluidity in brush-border membranes prepared from distal colonocytes of rats administered this agent for 5, 10 and 15 weeks, i.e., before the development of colon cancer. Furthermore, changes in the sphingomyelin content and sphingomyelin/phosphatidylcholine molar ratio of these membranes appeared, at least partially, to be responsible for these fluidity alterations. In an attempt to elucidate the mechanism(s) involved in these dimethylhydrazine-induced lipid changes, in the present studies the activities of sphingomyelin synthetase and magnesium-dependent neutral sphingomyelinase, enzymes involved in the synthesis and degradation of this phospholipid, respectively, were examined and compared in distal colonic brush-border membranes prepared from rats after 5, 10 or 15 weeks administration of dimethylhydrazine or diluent. The results of these studies demonstrate that alterations in both these enzymatic activities can be detected after administration of dimethylhydrazine and appear to, at least in part, be responsible for the changes in membrane sphingomyelin composition noted previously. These results as well as a discussion of their possible serve as the basis for the present report. PMID- 3024723 TI - Effects of vanadate on water and electrolyte transport in rat jejunum. AB - The effect of vanadate (orthovanadate, VO4-) on water and ion transport was studied in rat jejunum. Water transport was tested by single-pass perfusion in vivo and ion fluxes by the Ussing-chamber technique in vitro. The results suggest that vanadate has two actions on ion and water transport: At low concentrations (10(-4) M) it causes Cl-, Na+ and water secretion by stimulation of adenylate cyclase; At higher concentrations (10(-3) and 10(-2) M) it decreases net absorption of Na+ and Cl- by inhibition of (Na+ + K+)-ATPase. PMID- 3024724 TI - Escherichia coli ribosomal protein S1 does not protect the 49-nucleotide 3' terminal cloacin fragment of 16 S rRNA from nuclease S1. AB - Footprinting of ribosomal protein S1 on the 49-nucleotide 3' terminal cloacin DF13 fragment of 16 S rRNA at physiological ionic strength, pH and temperature yielded no detectable protection of any nucleotides from subsequent attack by the single strand specific nuclease S1, even at large excesses of ribosomal protein S1. PMID- 3024725 TI - NH2-terminal spin labeling of human hemoglobin--spin states and temperature dependence. AB - In order to explore fully how ligand- and temperature-induced alterations in the spin states of heme iron are related to protein readjustments, the spin label 4 isothiocyanate (I) was covalently attached at beta-93 cysteines and at NH2 terminal valines of various heme-iron ligand forms of human hemoglobin. It was found that the mobility of NH2-terminally bound spin labels depends on the magnetic moment of the heme iron. There is a an approximately linear relationship between the magnetic moment of the heme iron and the mobility of NH2-terminally bound spin labels. In accordance with our previous results, the temperature dependence of ESR spectra of spin-labeled hemoglobin suggests the temperature induced protein conformational change in those heme-iron ligand forms that are characterized by the equilibrium of the spin states of the heme iron. The conformational change was sensed at both spin-label-binding sites: at beta-93 cysteines and at NH2-terminal valines. PMID- 3024726 TI - The effect of oxidants on neutrophil-mediated degradation of glomerular basement membrane collagen. AB - The contribution of activated oxygen species to neutrophil-mediated degradation of basement membrane collagen was investigated. In preliminary experiments, pre exposure of either albumin or glomerular basement membrane to neutrophil myeloperoxidase with H2O2 and chloride increased their susceptibility to proteolysis 2-3-fold. In the basement membrane model, neutrophils are stimulated by trapped immune complexes to adhere, produce oxidants and degranulate. Degradation, measured as the amount of hydroxyproline solubilised, was due to neutral proteinases, particularly elastase, and depended on cell number and the amount of proteinase released. Experiments with oxidant scavengers and inhibitors and with neutrophils from donors with chronic granulomatous disease or myeloperoxidase deficiency showed that oxidants did not affect degradation of the basement membrane when this was measured on a per cell basis. However, oxidative inactivation of the released granule enzymes occurred. Activities of elastase, beta-glucuronidase and lysozyme were 1.5-2-times higher in the presence of catalase, but were unaffected by superoxide dismutase or hydroxyl radical scavengers. Inactivation did not occur with chronic granulomatous disease or myeloperoxidase deficient neutrophils. When related to the activity of released elastase, or to other degranulation markers, collagen degradation was decreased in the presence of catalase, or with chronic granulomatous disease or myeloperoxidase deficient cells. This implies that the basement membrane was made more digestible by myeloperoxidase-derived oxidants, as occurred in the cell-free experiments. Taken together, the results indicate that neutrophil oxidants have two opposing effects. They increase the susceptibility of the collagen to proteolysis and inactivate the proteinases responsible. PMID- 3024728 TI - The effect of dibutyryl cyclic AMP on the excretion of taurocholic acid from isolated rat liver cells. AB - Dibutyryl cyclic AMP (50-1000 microM) was found to increase the initial rate of efflux of taurocholic acid from isolated rat hepatocytes. Efflux of the bile acid was inhibited by sodium, and in the absence of sodium dibutyryl cyclic AMP failed to stimulate the rate. Increasing the concentration of calcium from 0 to 1.2 mM had no effect on the initial rate of taurocholic acid efflux from the cells, but the absence of calcium markedly reduced the effect of dibutyryl cyclic AMP. The results suggest that changes in the fluxes of sodium and calcium are involved in the effect of the cyclic nucleotide on taurocholic acid efflux from the cells. PMID- 3024727 TI - Cytoplasmic pH regulation in activated human neutrophils: effects of adenosine and pertussis toxin on Na+/H+ exchange and metabolic acidification. AB - When stimulated, neutrophils undergo a complex change in cytoplasmic pH (pHi): an incipient acidification, followed by an alkalinization which is due to activation of Na+/H+ exchange. When the latter is inhibited by amiloride or by removal of extracellular Na+, the actual magnitude of the initial acidification can be fully appreciated. The acidification is thought to be of metabolic origin, but the precise origin of the H+ (equivalents) remains undefined. We used adenosine, a modulator of neutrophil responsiveness, to identify the source of metabolic acid in cells stimulated by either formylmethionylleucylphenylalanine (fMet-Leu-Phe) or 12-O-tetradecanoylphorbol 13-acetate (TPA). Pretreatment of the cells with adenosine inhibited the fMet-Leu-Phe-induced respiratory burst, but secretion of specific and azurophilic granules, as well as aggregation were unaffected. In fMet-Leu-Phe-treated cells, adenosine reduced the acidification recorded in Na+ free media, but had no effect on the activation of the Na+/H+ antiport. Adenosine had little or no effect on the TPA-induced responses, including the pHi changes. The respiratory burst, as well as the cytoplasmic acidification were also inhibited in parallel by pretreating the cells with 'islet-activating protein' from Bordetella pertussis. It was concluded that activation of the NADPH-oxidase and/or the associated stimulation of the hexose monophosphate shunt play a major role in the metabolic acidification of stimulated neutrophils. PMID- 3024729 TI - Changes in myoblast membrane order during differentiation as measured by EPR. AB - The events which make possible the characteristic fusion of the cell membranes of embryonic myoblasts are known to involve modification of the cell membrane (Hausman, R.E., Dobi, E.T., Woodford, E.J., Petrides, S., Ernst, M. and Nichols E.B. (1986) Dev. Biol. 113, 40-48). Myoblasts from chick embryos were allowed to differentiate in gyrotory aggregate culture and the order of their membranes was measured by EPR. Two spin-labels which insert at different depths into the lipid bilayer were used. Measurement with the 5-nitroxystearate label showed an increase in myoblast membrane order (2T' parallel) from 0-15 h of culture and again from 26-38 h of culture. Measurement with the 12-nitroxystearate label showed the 0-15 h increase in order but the second increase was greatly reduced and shifted in time. While the specific sources of these changes in membrane order cannot yet be identified, the changes observed correlated well with known events of myogenic differentiation in vitro. The initial increase in membrane order occurred while the myoblasts were recovering from the effects of trypsin dissociation and undergoing gyrotory aggregation. The second increase in membrane order occurred during the known period of prostaglandin receptor activity and increased cell-cell adhesion. PMID- 3024730 TI - Enhanced uptake of carnitine by perfused rat liver following starvation. AB - Previously, the release of carnitine from the perfused rat liver was found to be protein-mediated, dependent on the nutritional state but not on metabolic energy. Further, it was shown to exceed the physiological demand by about 10-fold (Sandor et al. (1985) Biochim. Biophys. Acta 835, 83-91). In the present study the uptake of carnitine by perfused rat liver has been investigated. The liver tissue and the perfusate were in equilibrium when the carnitine concentration in the perfusate was close to 45 microM, physiological in the rat plasma. Under this condition, when no net carnitine transport occurred, an unidirectional uptake of L-[3H]carnitine was observed. Quantitatively, the uptake rate was 355 +/- 60 (S.D.) nmol/h per 100 g body weight at 45-50 microM perfusate concentration. This uptake capacity balances the previously reported excessive release (Sandor et al., op. cit.). On this basis we propose that a futile release/uptake cycle operates in carnitine transport across the liver cell membrane. Liverse of 24-h starved rats took up L-[3H]carnitine at 56% higher rate from the perfusate (75 microM) than livers of fed rats. Kinetic analysis revealed that fasting caused a decrease in Km value from 4.22 mM to 2.59 mM, whereas Vmax remained practically unchanged, average 0.95 mumol/min per 100 g body weight. D-[3H]Carnitine was transported at the same rate as L-carnitine and underwent the effect of fasting as well. The uptake was partially inhibited by 1 mM 2,4-dinitrophenol and 5 mM KCN, showing its dependency on metabolic energy. If Li+ replaced Na+ a strong inhibitory effect (to 20% of control) was observed, which suggests a co-transport of carnitine with Na+. Mersalyl, an SH reagent, had no effect on the uptake, whereas it practically abolished the release of carnitine from the perfused livers. This observation suggests that the inward and outward transport of carnitine are mediated by two different proteins. PMID- 3024731 TI - A covalent complex between horse heart cytochrome c and yeast cytochrome c peroxidase: kinetic properties. AB - The kinetic properties of a 1:1 covalent complex between horse-heart cytochrome c and yeast cytochrome c peroxidase (ferrocytochrome-c:hydrogen-peroxide oxidoreductase, EC 1.11.1.5) have been investigated by transient-state and steady state kinetic techniques. Evidence for heterogeneity in the complex is presented. About 50% of the complex reacts with hydrogen peroxide with a rate 20-40% faster than that of native enzyme; 20% of the complex exists in a conformation which does not react with hydrogen peroxide but converts to the reactive form at a rate of 20 +/- 5 s-1; 30% of the complex does not react with hydrogen peroxide to form the oxidized enzyme intermediate, cytochrome c peroxidase Compound I. Intramolecular electron transfer between covalently bound ferrocytochrome c and an oxidized site in cytochrome c peroxidase Compound I is too fast to measure, but a lower limit of 600 s-1 can be estimated at 5 degrees C in a 10 mM potassium phosphate buffer at pH 7.5. Free ferrocytochrome c reduces cytochrome c peroxidase Compound I covalently bound to ferricytochrome c at a rate 10(-4) to 10(-5)-times slower than for free Compound I. The transient-state ferrocytochrome c reduction rates of Compound I covalently linked to ferricytochrome c are about 70-times too slow to account for the steady-state catalytic properties of the 1:1 covalent complex. This indicates that hydrogen peroxide can interact with the 1:1 complex at sites other than the heme of cytochrome c peroxidase, generating additional species capable of oxidizing free ferrocytochrome c. PMID- 3024732 TI - The electronic and magnetic properties of rubredoxin: a low-temperature magnetic circular dichroism study. AB - Oxidized rubredoxin from Clostridium pasteurianum has been investigated by magnetic circular dichroism (MCD) spectroscopy over the temperature range 1.5 to 150 K and at magnetic fields between 0 and 4.5 tesla. The results show that studies of the temperature and field dependence of MCD transitions afford insight into the polarization of electronic transitions for ground states with large g value anisotropy, in addition to estimates of ground-state g values and zero field splitting parameters. In agreement with the assignment made by Eaton and Lovenberg (Eaton, W.A. and Lovenberg, W. (1973) in Iron-Sulfur Proteins, Vol. II (Lovenberg, W., ed.), pp. 131-162, Academic Press, New York), the ultraviolet visible spectrum of oxidized rubredoxin is assigned to two S----Fe(III) charge transfer transitions (both 6A1----6T2 under tetrahedral symmetry), each spanning a range of 650-430 nm and 430-330 nm, respectively. The observed splitting in each of these transitions is attributed to a predominant axial distortion in the excited state resulting in effective D2d symmetry. PMID- 3024733 TI - Spectroscopic studies of the seven-iron-containing ferredoxins from Azotobacter vinelandii and Thermus thermophilus. AB - The seven-iron-containing ferredoxins from Azotobacter vinelandii and Thermus thermophilus have been investigated by low-temperature magnetic circular dichroism (MCD) and electron paramagnetic resonance (EPR) spectroscopies and room temperature ultraviolet-visible absorption spectroscopy. The results confirm the presence of one trinuclear and one tetranuclear iron-sulfur cluster in both ferredoxins and facilitate comparison of the electronic and magnetic properties of the oxidized and reduced [3Fe-xS] clusters. MCD magnetization data are consistent with an S = 2 ground state for both reduced [3Fe-xS] clusters, but indicate differences in the rhombicity of the zero-field splittings. The data permit rationalization of the absence of a delta M = 4 EPR transition for the reduced [3Fe-xS] cluster in A. vinelandii ferredoxin I. Spectroscopic studies of anaerobically isolated A. vinelandii ferredoxin I do not support the hypothesis that the [3Fe-xS] cluster arises as a result of aerial oxidative damage to a [4Fe 4S] cluster during isolation. The possibility that two distinct forms of [3Fe-xS] clusters can exist in A. vinelandii ferredoxin I was investigated by spectroscopic studies as a function of pH. The results reveal two distinct and interconvertible forms of the reduced [3Fe-xS] cluster, but do not permit rationalization of the inconsistencies in the structural data that have been reported for the oxidized clusters. PMID- 3024734 TI - Purification and some properties of peroxidases of rat bone marrow. AB - Myeloperoxidase and eosinophil peroxidase were separated and purified from rat bone marrow cells using cetyltrimethylammonium bromide as the solubilizer and then with column chromatographies on CM-Sephadex C-50 and Con A-Sepharose. Both purified enzymes were observed to be apparently homogeneous by SDS-polyacrylamide gel electrophoresis. Myeloperoxidase consisted of two subunits of Mr 57,000 and 15,000, and eosinophil peroxidase two of 53,000 and 14,000. On structural analysis of the enzymes, their visual and ESR spectra revealed that the structure surrounding the heme in myeloperoxidase was different from that in eosinophil peroxidase. Moreover, substrate specificity and sensitivity to inhibitors such as azide and cyanide differed between the two enzymes. Rat bone marrow possesses two distinct peroxidases, myeloperoxidase and eosinophil peroxidase, which have different subunits and different heme microenvironments. Therefore, the difference in enzymatic function between the two peroxidases may be due to their structures. PMID- 3024735 TI - Glucocorticoid-induced elevation of serum high-density lipoprotein-cholesterol and its reversal by adrenocorticotropin in the rat. AB - The effect of administration of a high dose of glucocorticoid (triamcinolone) on serum lipids and lipoproteins was studied in rats. Changes in serum lipids, especially cholesterol, were most marked when 5 mg/kg body weight of triamcinolone was injected daily for 5 days. Serum lipoproteins were separated by ultracentrifugation followed by gel-filtration chromatography. Cholesterol distribution between apolipoprotein B-containing lipoproteins (very-low-density and low-density lipoproteins), high-density lipoprotein1 (HDL1), and HDL2 was determined after administration of triamcinolone with or without additional treatment with adrenocorticotropin (ACTH; Cortrosyn, 6 IU/rat). When triamcinolone was administered, cholesterol concentrations in HDL1 and HDL2 were elevated in a dose-dependent manner, but there was no significant change in apolipoprotein B-containing lipoprotein cholesterol levels. When ACTH was administered in combination with triamcinolone, the concentrations of all serum lipids except triacylglycerol were significantly lowered compared with rats treated with triamcinolone alone. HDL1-cholesterol concentration in serum was significantly (P less than 0.001) lowered from 69 +/- 13 mg/dl (mean +/- S.D.) in triamcinolone-treated rats to 36 +/- 4 mg/dl by the administration of ACTH plus triamcinolone. The additional administration of ACTH in triamcinolone-treated rats caused a slight, but significant, decrease in cholesterol concentration in apolipoprotein B-containing lipoproteins; however, HDL2-cholesterol level was not significantly affected, although there was a tendency for it to be lowered. PMID- 3024736 TI - Phosphoinositide turnover enhanced by angiotensin II in isolated rat glomeruli. AB - To clarify the signal transduction mechanism of angiotensin II in renal glomeruli, we studied the effect of the hormone on phospholipid metabolism using isolated rat glomeruli. Stimulation of the glomeruli pulse-chase labeled with [3H]glycerol by angiotensin II caused a rapid (within 15 s) breakdown of phosphatidylinositol 4,5-bisphosphate (PIP2) with a concurrent production of 1,2 diacylglycerol. This effect of angiotensin II was in a dose-dependent manner within the range from 10(-12) M to 10(-6) M, and was inhibited by saralasin. Angiotensin II also decreased the 3H radioactivity of PIP slightly only at 15 s and increased that of phosphatidic acid after 15 s, with no significant effect upon the labelings of phosphatidylinositol (PI), phosphatidylcholine (PC) and phosphatidylethanolamine (PE) within 1 min. The change in phospholipid metabolism by angiotensin II was similar when the glomeruli were labeled with [32P]orthophosphate: the decrease in the labeling of PIP2 and the increase in the labeling of phosphatidic acid after 15 s. In addition, 32P labeling of PI increased after 2 min. These results suggest that angiotensin II, after binding to glomerular receptors, induces initial PIP2 hydrolysis to diacylglycerol and subsequent resynthesis of PIP2 through phosphoinositide turnover. PMID- 3024737 TI - Production of inositol trisphosphates and inositol tetrakisphosphate in stimulated pancreatic islets. AB - Glucose and carbamylcholine caused concentration-dependent increases in the production of total [3H]inositol phosphates in [3H]inositol-labelled rat pancreatic islets. When extracts from islets stimulated with glucose, carbamylcholine or depolarising concentrations of K+ were analysed using anion exchange high performance liquid chromatography, increased production of [3H]Ins1,4,5-P3 was detected, and in addition, elevated levels of two other labelled compounds which co-chromatographed with Ins1,3,4-P3 and Ins1,3,4,5-P4. In the case of carbamylcholine and high K+, such an effect was apparent within 20 s, whereas glucose appeared to cause a delayed response. In the presence of 5 mM LiCl, the accumulation of Ins1,3,4-P3 was more marked. The presence of LiCl had no major influence on the levels of Ins1,4,5-P3 or Ins1,3,4,5-P4. It is suggested that the stimulation of pancreatic islets with glucose, carbamylcholine or high K+ results in the hydrolysis of inositol lipids with the production of Ins1,4,5 P3 and in addition, Ins1,3,4-P3 and Ins1,3,4,5-P4. The physiological functions of these novel inositol phosphates in islets remain to be established. PMID- 3024738 TI - Studies on the mechanism of interleukin 1 stimulation of platelet activating factor synthesis in human endothelial cells in culture. AB - Interleukin 1 promotes the conversion of the biologically inactive lyso-platelet activating factor (lyso-PAF) to the bioactive platelet activating factor (PAF) by an acetylation reaction in cultured human endothelial cells. After 2 h stimulation with interleukin 1, 1-O-alkyl-2-lysoglycero-3-phosphocholine (GPC): acetyl CoA acetyltransferase is activated, reaching a plateau after 6 h and then declining to the basal value within 24 h. This time course is comparable to that of PAF production. These cells are able to incorporate [3H]acetate and [3H]lyso PAF into PAF. Synthetized [3H]PAF is then catabolized in [3H]alkylacyl phosphoglycerides. 1-O-alkyl-2-acetylglycerol: CDP-choline cholinephosphotransferase and 1-O-alkyl-2-acetyl-GPC: acetylhydrolase activities are both present in endothelial cells, but are not activated under our conditions of stimuli. These findings indicate that interleukin 1 induces the PAF synthesis by a deacylation/reacetylation mechanism into human endothelial cells. PMID- 3024739 TI - The synthesis and turnover of 5'-nucleotidase in primary cultured hepatocytes. AB - The synthesis and degradation of 5'-nucleotidase has been studied in rat hepatocytes. Primary cultures of rat hepatocytes were established with the cells showing evidence of polarity after 24-36 h in culture. After a 30 h lag period 5' nucleotidase activity increased to a plateau level similar to the activity found in whole liver. The half life of the enzyme after reaching the plateau of activity was 22.8 h. Pulse-chase biosynthetic labelling studies of 5' nucleotidase in the cultured hepatocytes using [35S]methionine showed that the 5' nucleotidase monomer was synthesised as an Mr 67,000 form which was converted to the mature Mr 72,000 form. [35S]Methionine labelling studies in the presence of tunicamycin showed that the unglycosylated protein monomer was an Mr 57,000 form. The immature Mr 67,000 form of 5'-nucleotidase was sensitive to endoglycosidase H, whereas the mature form was sensitive only to endoglycosidase F. The data presented are consistent with 5'-nucleotidase in a polarised cell being synthesised and processed like other membrane glycoproteins, in contrast to earlier reports. PMID- 3024740 TI - Oncogenes and the control of cell growth: fitting together the pieces of the puzzle. PMID- 3024741 TI - The opioid peptides and their receptors. PMID- 3024742 TI - A single gene produces multiple sericin messenger RNAs in the silk gland of Bombyx mori. AB - The sericins are a family of major cocoon proteins specifically synthesized in the middle silk gland of the silkworm Bombyx mori. The 5' part of one sericin gene had been cloned and described by Okamoto et al. (1982, J. Biol. Chem. 257, 15192-15199). Using a differential screening procedure of Bombyx genomic libraries, we obtained the 3' part of this gene. We demonstrate that it consists of a single gene extending over 24 kb, present in two allelic forms in hybrid strains. This gene encodes for four mRNAs which result from a unique transcript by an alternative splicing mechanism. This explains, at least partially, the diversity of the sericins found in the cocoon. PMID- 3024744 TI - The Na+/H+ antiport of eukaryotic cells: relationship between the kinetic properties of the system and its physiological function. AB - The Na+/H+ antiport is present in the plasma membrane of virtually all vertebrate cells and it plays a central role in cell homeostasis. The pharmacological properties and the characteristics of the interaction of extracellular Na+, Li+, H+ and of intracellular H+ with the Na+/H+ antiport are reviewed herein. The kinetic properties of the system are shown to be essential for defining its four main physiological functions: transepithelial ion transport, control of the pHi, control of the intracellular Na+ concentration, and control of the cell volume. The activity of the Na+/H+ antiport can be modulated by a large number of effectors which are thought to act via protein kinases. At least three mechanisms of activation of the Na+/H+ exchanger are defined from the analysis of the kinetic properties of the system. Activation of the Na+/H+ antiport leads to very different consequences, depending upon the activity of other ion transporting systems in the membrane. PMID- 3024743 TI - Phospholipid dependence of the neutral sphingomyelinase in rat liver plasma membranes. AB - Investigations have been carried out on the influence of the phospholipid composition of rat liver plasma membranes and of their physico-chemical properties on the activity of membrane-bound neutral sphingomyelinase. The membrane phospholipid composition was modified by the incorporation of different phospholipids into the membrane bilayer by means of lipid transfer proteins, n butanol delipidation or exogenous sphingomyelinase (Staphylococcus aureus) treatment. The results indicate that the activity of neutral sphingomyelinase in liver plasma membranes depends upon phosphatidyl choline presence in the membrane bilayer and not upon membrane fluidity. PMID- 3024745 TI - Gastric (H+,K+)-ATPase. AB - Gastric acid secretion results from the activity of a specific ATPase, the (H+,K+)-ATPase. This enzyme, discovered in 1973, exchanges H+ for K+. It has two ATP binding sites, both involved in enzyme activity, whose affinities vary as a function of the H+ and K+ concentrations. Hydrolysis of ATP at the highest affinity site leads to the synthesis of a covalent aspartyl phosphate which accumulates in the absence of K+. The presence of this cation accelerates dephosphorylation resulting in the stimulation of ATPase (and PNPPase) activity. The structure of membranous (H+,K+)-ATPase is poorly defined. n-Octylglucoside solubilizes an active enzyme of 390-420 kDa which can be partly depolymerized using cholate. The monomer, characterized in SDS has a 95 kDa molecular mass and is inactive. In the presence of magnesium, (H+,K+)-ATPase catalyzes the active and neutral exchange of H+ for K+ at the expense of ATP. In the absence of ATP, (H+,K+)-ATPase acts as a passive transporter exchanging K+ for K+ at maximal rate and H+ for K+ at a 20 times slower rate. PMID- 3024746 TI - Cerebrospinal fluid levels of angiotensin-converting enzyme, acetylcholinesterase, and dopamine metabolites in dementia associated with Alzheimer's disease and Parkinson's disease: a correlative study. AB - Mean levels of the two hydrolases angiotensin-converting enzyme (ACE) and acetylcholinesterase (AChE), the dopamine metabolites dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA), and total protein concentration were examined in cerebrospinal fluid (CSF) samples from a group of patients with dementia of the Alzheimer's type, a group of comparably demented patients with Parkinson's disease, and a neurologically healthy elderly control group. Both pathological groups exhibited a significant decrease in the mean levels of ACE activity and DOPAC per milliliter and were distinguishable from one another based on mean CSH HVA levels. Unlike the Parkinson's disease group, whose mean concentration of HVA was lower than, but not significantly different from that of the control group, the mean HVA concentration of the Alzheimer's disease group was significantly elevated. In contrast, comparisons of the mean CSF AChE activity (expressed per milliliter or per milligram of protein) and CSF total protein concentration did not reveal significant differences for any of the groups. Independent of CSF protein concentration, ACE activity per milliliter exhibited a positive correlation with AChE activity per milliliter within the control and Parkinson's disease groups, whereas a statistically significant correlation for these CSF hydrolases was not observed within the Alzheimer's disease group. Thus, the CSF profiles for patients with mild dementias associated with Alzheimer's or Parkinson's disease differed by at least two neurochemical criteria. Based on the levels of ACE activity, DOPAC, and HVA per milliliter of CSF, two discriminant functions were derived and resulted in the correct classification of 71% of all subjects (n = 38) into Alzheimer's disease, Parkinson's disease, and neurologically healthy control groups. PMID- 3024747 TI - Development of opiate ([3H] naloxone)-binding sites in female rabbit brain: correlation with prepubertal gonadotropin secretion. AB - In neurochemical terms, little is known concerning the control of puberty onset in the female rabbit. In view of the established involvement of brain opiates in the sexual maturation of the rat, we have investigated the prepubertal development of opiate-binding sites in the hypothalamus and cerebral cortex of the rabbit. Binding of the opiate antagonist, [3H] naloxone, to thick (350 micron) slices of rabbit brain was found to be reversible, stereospecific, saturable, and of high affinity. In all respects these sites possessed the characteristics of opiate receptors. Specific binding (Bmax and KD) values were determined at 1, 8, 29, 40, 51, 100, and 168 days after birth in the hypothalamus and cerebral cortex. All all ages binding in the hypothalamus was higher, per mg of tissue, than in the cortex. Major differences in the pattern of development were also evident. In the cortex the Bmax slowly increased from a minimum at Day 1 to a maximum at about 100 days when puberty normally occurs. In contrast, binding in the hypothalamus rose rapidly to a maximum at 40 days and then fell abruptly, by about 40% at Day 51, after which a slow increase through puberty took place. This peak in the hypothalamic Bmax value correlates closely with the major prepubertal surge of gonadotropin secretion. It remains to be determined whether the coincidence of spontaneous gonadotropin secretion with the rapid appearance of hypothalamic opiate receptors is developmentally meaningful for the reproductive system. PMID- 3024748 TI - Proteolysis by collagenase preparations alters the macromolecular composition of the porcine zona pellucida. AB - The zona pellucida (ZP) from pig oocytes was isolated using two different methods. In the first method, the ZP was isolated using sieving procedures. In the second method, an enzymatic step with collagenase was used in addition to sieving procedures. Several commercial collagenase preparations were tested. The macromolecular composition of the ZP isolated by these two methods was determined by two-dimensional polyacrylamide-gel electrophoresis after disulfide bond reduction. The ZP prepared by the sieving method contained four glycoprotein families with apparent molecular weights of 25,000, 55,000, 65,000, and 90,000. The ZP obtained using the enzymatic method was distinctly different, lacking the highest molecular-weight family (90,000) and containing at least three new constituents with apparent molecular weights of 70,000, 40,000, and less than 25,000. Commercial collagenase preparations, when analyzed by two-dimensional polyacrylamide-gel electrophoresis to assess homogeneity, contained numerous protein components. The trypsin-like protease concentration in the collagenase preparations was determined to be 3.4-42 X 10(-8) M as determined by activity measurement using benzoyl-DL-arginine-beta-naphthylamide as substrate or the active site titrant p-nitrophenyl-p'-guanidinobenzoate. Thus, the ZP prepared by the enzymatic method, using collagenase preparations, had an altered macromolecular composition, thereby rendering the ZP unsuitable for most structural, immunological, or sperm-binding studies. PMID- 3024749 TI - Adhesion of carboxylate cements to hydroxyapatite. I. The effect of the structure of aliphatic carboxylates on their uptake by hydroxyapatite. AB - An investigation was made into the effect of the structure and functionality of aliphatic carboxylates on their sorption onto hydroxyapatite with a view to ascertaining factors affecting the adhesion of polyacrylate cements to tooth materials. In general, the amount of carboxylate sorbed was found to increase with the number of groups contained in the molecule. Thus, polyacrylate was found to be much more strongly sorbed than low molecular weight species. Sorption of the low molecular weight species appeared to be related to stereochemical factors rather than to the stability constants of their calcium chelates. PMID- 3024750 TI - An in vitro study of the role of dentine in moderating the cytotoxicity of zinc oxide eugenol cement. AB - The protective role of different dentine fractions and of dentine slices in moderating the cytotoxicity of zinc oxide eugenol (ZOE) was investigated. The collagen fraction of dentine powder provided increased protection over intact powder. Slices of dentine offered greater protection probably by providing a physical barrier to the diffusion of eugenol which may also bind to the contents of the dentine tubules. With increasing thickness of the dentine slices this protection was increased. ZOE stimulated calcium release from dentine but in view of the low levels attained it is unlikely that this process has any significant effect on the protective role of dentine. PMID- 3024751 TI - The gap junction channel. Its aqueous nature as indicated by deuterium oxide effects. AB - The effects of temperature and solvent substitution with deuterium oxide (D2O) on axoplasmic (ga) and gap junctional (gj) conductances were examined in the earthworm septate median giant axon (MGA). The temperature coefficients (Q10) for ga and gj were 1.4 and 1.5, respectively, between 5 and 15 degrees C. Substitution with D2O rapidly reduced both ga and gj by 20% and increased the Q10's to 1.5 and 1.8, respectively. The reduction in ga upon substitution with D2O and with cooling in either solvent reflects the changes that occur in solvent viscosity, which indicates that ion mobility in axoplasm, as in free solution, is primarily governed by viscous properties of the solvent. The similar initial reduction observed for gj suggests that solvent occupies the gap junction channel volume and influences transjunctional ion mobility. With time there was a further reduction in gj at 20 degrees C and a larger Q10 in D2O. The enhanced effects of D2O on gj cannot be accounted for by solvent viscosity alone and may be due to an increased hydration of the channels and/or the transport ions and by isotope effects of hydrogen-deuterium exchange on the channel protein that reduce gj. PMID- 3024752 TI - Molecular orbital calculations on the oligopeptides netropsin, distamycin and related compounds. PMID- 3024753 TI - The effect of monovalent cations on the association behavior of guanosine 5' monophosphate, cytidine 5'-monophosphate, and their equimolar mixture in aqueous solution. PMID- 3024754 TI - Evaluation of the effects induced by four opiate drugs, with different affinities to opioid receptor subtypes, on anterior pituitary LH, TSH, PRL and GH secretion and on cortisol secretion in normal men. AB - In order to ascertain the subtype(s) of opioid receptors involved in the control of pituitary function the effects of four different opiate drugs (morphine, pentazocine, nalorphine and buprenorphine) were studied in four groups of six normal male volunteers. Each of the drugs tested induced, with varying degrees, both a significant increase in PRL and a significant decrease in LH and cortisol. On the contrary TSH secretion was stimulated by buprenorphine and morphine only and GH by nalorphine only. FSH did not change significantly after administration of any drug. Taking into account the different affinities of the drugs used by us for the different opioid receptors, our data do not allow to demonstrate a well defined correlation between subtypes of opioid receptors and the control of pituitary hormone secretion. The possible explanations of this fact are discussed. PMID- 3024755 TI - Loss of interleukin-2 requirement for the generation of T colonies defines an early event of human T-lymphotropic virus type I infection. AB - Accessory cells and/or soluble factors, together with interleukin 2 (IL2), are required for the proliferation and differentiation of phytohemagglutinin (PHA) activated T lymphocytes. Human T-lymphotropic virus, type I (HTLV-I), a human retrovirus isolated from patients with adult T cell leukemia, can transform T cells in vitro. We investigated the role of HTLV-I-transformed T cell lines as accessory cells in promoting the growth of T colony-forming cells. We found that T cells isolated by E rosetting and then activated with PHA, when seeded with as few as 5 X 10(3) irradiated HTLV-I-producing cells, could generate colonies in the absence of IL2. We analyzed further the effects of HTLV-I virions on T colony formation. Infection of T cells with semipurified HTLV-I viral particles promoted colony formation, in the absence of IL2, of accessory cells or soluble factors. The same results were obtained either with monocyte-depleted T lymphocytes, or with T4 or T8 lymphocytes. Furthermore, T lymphocytes in the presence of heat inactivated HTLV-I (devoid of replicative potential) could form colonies independently of IL2. Finally, experiments with sera positive for HTLV-I antibodies (to abolish binding of viral particles to cellular receptors) indicated that HTLV-I promoted IL2-independent colony formation, only by "touching" T colony-forming cells. These results taken together demonstrate that the loss of the exogenous IL2 (and other growth-helping factors) requirement defines an early event of HTLV-I infection. The results also suggest that viral attachment to T cells possibly supplies an accessory function triggering autocrine secretion of IL2 by these cells. PMID- 3024756 TI - Characterization of the inhibition of tissue factor in serum. AB - Tissue factor (TF) is a lipoprotein cofactor that markedly enhances the proteolytic activation of factors IX and X by factor VIIa. The functional activity of TF is inhibited by serum in a time- and temperature-dependent fashion. The inhibitory effect is also dependent on the presence of calcium ions and can be reversed by calcium chelation (EDTA) and dilution, thus excluding direct proteolytic destruction of TF as the mechanism for inhibition. Using crude TF, serum immunodepleted of factor VII, and serum depleted of the vitamin K dependent coagulation factors by BaSO4 absorption, it is shown that TF factor inhibition requires the presence of VII(a), X(a), and an additional moiety contained in barium-absorbed serum. When each of the other required components were at saturating concentrations, half-maximal inhibition of TF occurred in reaction mixtures containing 2% (vol/vol) of TF at a factor VII(a) concentration of 4 ng/mL (80 pmol/L), a factor X concentration of 50 ng/mL (850 pmol/L), and a concentration of barium-absorbed serum of 2.5% (vol/vol). Catalytically active factor Xa appeared to be required for the generation of optimal TF inhibition. The results are consistent with the conclusions of Hjort that barium-absorbed serum contains a moiety that inhibits the VIIa-Ca2+-TF complex. The role of factor X(a) in the generation of the inhibitory phenomenon remains to be elucidated. The inhibitor present in serum (plasma) may in part be produced by the liver in vivo since cultured human hepatoma cells (HepG2) secrete this inhibitory activity in vitro. PMID- 3024758 TI - Steady state levels of factor X mRNA in liver and Hep G2 cells. AB - In order to examine the control of human factor X biosynthesis we have molecularly cloned the cDNA and investigated the expression of the Factor X gene. A recombinant clone of approximately 1100 base pairs in length containing the sequence of factor X was identified in a lambda gt11 human liver cDNA library by screening with polyclonal antibodies. One plaque was selected and confirmed for specificity with a mixture of five factor X specific monoclonal antibodies (MoAbs). A partial nucleic acid sequence of the 5' end of the cDNA corresponded to the described amino acid sequence between residues 41 and 56 of the light chain of factor X. Northern blot analysis of RNA from human liver and the hepatoma cell line, Hep G2, identified the factor X mRNA as a single molecular species of approximately 1700 bases. Cell lines which do not secrete factor X did not contain factor X mRNA indicating restriction of transcription to hepatocytes. Slot-blot hybridization analysis of factor X and actin mRNA demonstrated no change in the levels of total or specific factor X mRNA in Hep G2 cells following treatment with warfarin or vitamin K. We conclude that modulation of factor X production by these drugs, known to influence gamma-carboxylation and total factor X secretion by these cells, is mediated by changes in posttranscriptional events rather than by effects on the steady state levels of factor X mRNA. PMID- 3024757 TI - Human GM-CSF primes neutrophils for enhanced oxidative metabolism in response to the major physiological chemoattractants. AB - Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a T cell-derived lymphokine which induces hematopoietic precursor cells to proliferate in vitro and differentiate to neutrophils and macrophages. GM-CSF also inhibits the motility of mature neutrophils (NIF-T activity), and primes neutrophils to enhance oxidative metabolism in response to the bacterial chemoattractant, N formyl-methionyl-leucyl-phenylalanine (f-MLP). The present study was designed to determine whether this lymphokine also enhances neutrophil oxidative metabolism in response to the other major physiological chemoattractants which include complement-derived C5a, and the 5-lipoxygenation product of arachidonic acid, leukotriene B4 (LTB4). Superoxide anion production was measured as superoxide dismutase-inhibitable cytochrome C reduction. Purified biosynthetic GM-CSF enhanced superoxide anion production by neutrophils in response to f-MLP, C5a desArg, and LTB4. In contrast to several other factors which prime neutrophils, GM-CSF did not prime for an enhanced oxidative response to phorbol myristate acetate (PMA). These results suggest that GM-CSF may be an endogenous regulator of neutrophil inflammatory responses induced by the major physiological chemoattractants. PMID- 3024759 TI - cis-Retinoic acid stimulates the clonal growth of some myeloid leukemia cells in vitro. AB - We studied the effects of cis-retinoic acid (cisRA) on the clonogenic growth of samples of leukemic cells from 35 patients with acute nonlymphocytic leukemia (ANLL). We observed significant inhibition of leukemic colony growth in 17 samples by 10(-7) to 10(-6)M cisRA. However, we found that retinoid exposure resulted in striking stimulation of clonal growth in ten samples at the same drug concentrations. With the exception of cases with promyelocytic features, there was no morphologic or functional evidence that cisRA induced the leukemic blasts to differentiate. Both inhibition and stimulation were dose-dependent and observable at pharmacologically achievable levels of cisRA. Leukemic cells with monocytic features more frequently demonstrated a stimulatory response than did those without monocytic features. Depletion of T lymphocytes and monocytes did not alter the type of growth response. Assays for cellular retinoic acid-binding protein (CRABP) were performed on five samples (two with inhibitory growth responses, two with stimulatory responses, and one with no growth) and failed to reveal detectable levels of CRABP in any case. The addition of cisRA to liquid suspensions of leukemic cells produced no significant change in the number of viable cells. We conclude that the effects of cisRA on leukemic colony growth are not cytotoxic and not mediated by T lymphocytes, monocytes, or CRABP. More importantly, cisRA appears to enhance the growth of certain human leukemia cells in vitro. Taking into account the increasing use of retinoids in clinical trials for patients with leukemia, the latter findings may represent a significant cautionary note. PMID- 3024760 TI - Immunogenic nature of a Pol gene product of HTLV-III/LAV. AB - The present studies were initiated to define the coding region of a 34 kilodalton (kd) protein (p34) frequently observed with antibodies from HTLV-III/LAV-infected people by immunoblotting and radioimmunoprecipitation (RIP) techniques. We have directly mapped this viral protein to the pol gene of HTLV-III/LAV by radiolabeled amino acid sequence analysis. This region at the 3' end of the pol gene is predicted to encode the endonuclease/integrase of the virus. The seroprevalence rate of antibodies to the pol gene products p64 and p53 and to the endonuclease p34 were evaluated. Of 161 HTLV-III/LAV seropositive people tested by immunoblotting procedures, greater than 98% had antibodies which reacted to p64/p53 and 92.6% reacted to p34 indicating that these viral proteins are highly immunogenic in nature. We have also analyzed the serum of nine healthy people living in West Africa who were infected with HTLV-IV, a closely related retrovirus. Nine of nine seropositive people had antibodies that cross-reacted to p34 of HTLV-III/LAV, whereas only seven of nine reacted to p64/p53. These studies and our earlier observations indicate that current diagnostic procedures for screening for HTLV-III/LAV infection may also detect HTLV-IV seropositive individuals, pointing to a need for more specific assay systems. PMID- 3024761 TI - Purification and properties of bacterially synthesized human granulocyte macrophage colony stimulating factor. AB - Human granulocyte-macrophage colony stimulating factor (GM-CSF) has been synthesized in high yield using a temperature inducible plasmid in Escherichia coli. The human GM-CSF is readily isolated from the bacterial proteins because of its differential solubility and chromatographic properties. The bacterially synthesized form of the human GM-CSF contains an extra methionine residue at position 1, but otherwise it is identical to the polypeptide predicted from the cDNA sequence. The specific activity of 2.9 X 10(7) units/mg of protein for purified bacterially synthesized human GM-CSF indicates that despite the lack of glycosylation, the molecule is substantially in its native conformation. This molecule stimulated the same number and type of both seven- and 14-day human bone marrow colonies as the CSF alpha preparation from human placental conditioned medium. Human GM-CSF had no activity on murine bone marrow or murine leukemic cells. There was no detectable, direct stimulation of adult human erythroid burst forming units (BFU-E) by the bacterially synthesized human GM-CSF. Although impure preparations containing native human GM-CSF (eg, human placental conditioned medium) stimulated the formation of mixed colonies, even in the presence of erythropoietin, the bacterially synthesized human GM-CSF failed to stimulate the formation of mixed colonies from adult human bone marrow cells. The bacterially synthesized human GM-CSF increased N-formyl-methionyl-leucyl phenylalanine (FMLP)-induced superoxide production and lysozyme secretion. Antibody-dependent cytotoxicity and phagocytosis by human neutrophils was stimulated by the bacterially synthesized human GM-CSF and eosinophils were also activated in the antibody-dependent cytotoxicity assay. PMID- 3024762 TI - Divalent cation-dependent and independent surface expression of thrombospondin on thrombin-stimulated human platelets. AB - Thrombospondin (TSP), a platelet alpha-granule protein, becomes expressed on the surface of thrombin-stimulated platelets. The surface expression of this protein occurs through two distinct mechanisms. At low platelet concentrations (1 X 10(8)/mL), a divalent ion-dependent, low-capacity mechanism predominates. At higher cell concentrations, a divalent ion-dependent, higher capacity mechanism prevails that can account for greater than 90% of all the TSP surface expression measured. This mechanism requires the presence of both calcium and magnesium (Ca + Mg). The dependence of the divalent ion-dependent surface expression on platelet concentration suggests that release of the molecule from the cell followed by its binding to the cell surface mediates this component of the endogenous TSP-platelet interaction. These data are consistent with a two receptor model for the platelet-surface expression of the endogenous TSP pool. PMID- 3024763 TI - The relationship of myeloperoxidase activity to neutrophil maturity and other hematologic indicators of infection. AB - Myeloperoxidase activity (MPO) in 100 male hospitalized patients was quantified by enzyme cytochemical flow cytometric analysis. These data were compared with commonly used laboratory indicators of infection. The correlation coefficients were statistically significant for increased MPO activity with band count, toxic granulation, and the Schilling Index; however, the orders of magnitude were too low to suggest the use of MPO as a clinical parameter. Patients were given an infection rank based on their numbers of abnormal laboratory indicators of infection. There was a highly significant difference in mean neutrophil peroxidase (MNP) activity between those patients with three or more indicators of infection and those with two or less: t = 3.68, p less than 0.001. It is concluded that whereas MNP may not be a useful indicator of a left shift or of leukocytosis, it might be helpful as an indicator of infection or inflammation. Further studies will be required to support this hypothesis. PMID- 3024765 TI - [Determination of herpes antibodies in the aqueous humor by the ELISA method in the diagnosis of stromal keratitis]. PMID- 3024764 TI - Effect of synthetic detergents on germination of fern spores. PMID- 3024766 TI - Inpatient group psychotherapy. AB - Psychiatry has available many different treatment methods. Characteristically, there is polarization between psychotherapeutic and medical approaches, seen as competing and incompatible. However, given flexibility they can be complementary and synergistic. This article reviews the place of inpatient group psychotherapy on the contemporary psychiatric ward. Its main theme is that group therapy is an important element in treatment, which to be effective should be central and not peripheral in the ward. PMID- 3024767 TI - Modification of radioresponse of intermediate and B-type spermatogonia by a phosphorothioate. PMID- 3024768 TI - Treatment of parotid gland tumours by conservative parotidectomy. PMID- 3024769 TI - Primary carcinoma of Stensen's duct. PMID- 3024770 TI - Metastases from hepatocellular carcinoma in sclerosed oesophageal varices in cirrhotic patients. PMID- 3024771 TI - Hepatocellular carcinoma in urban born blacks: frequency and relation to hepatitis B virus infection. AB - Chronic hepatitis B virus infection is far less common in urban born than in rural born southern African blacks, who also have a high incidence of hepatocellular carcinoma. A case-control study was carried out to determine the relative frequency of hepatocellular carcinoma and its relation to hepatitis B virus infection in urban born blacks. Three hundred and ninety two black patients with hepatocellular carcinoma and matched controls seen at two city hospitals were classified by questioning as urban born or rural born. The ratio of rural born to urban born blacks among the controls was 1.1:1.0 (207/185), whereas in the patients with cancer the ratio was 4.8:1.0 (324/68) (p less than 0.0001). Analysis of the prevalence of hepatitis B markers in 62 urban born and matched rural born blacks with hepatocellular carcinoma showed no differences in the frequency of current or past hepatitis B virus infection. It is concluded that urban born blacks are less likely than rural born blacks to develop hepatocellular carcinoma, but when they do the tumour is equally likely to be related to infection with hepatitis B virus. The findings lend further support to an important role for chronic hepatitis B virus infection in the aetiology of hepatocellular carcinoma. PMID- 3024772 TI - Hyponatraemia as a cause of reversible ataxia. PMID- 3024773 TI - Incidence of chickenpox in adults and recruitment of plasma donors for manufacture of zoster immunoglobulin. PMID- 3024774 TI - The excitation by neurotensin of nucleus tractus solitarius neurons induces apneustic breathing. AB - The possible involvement of neurotensin in the regulation of respiratory drive has been tested on single brainstem respiratory related neurons and on the global respiratory output. The neuropeptide was locally applied either by microiontophoresis or by pressure injection in the dorsal and ventral respiratory areas of the anesthetized bivagotomized cat. Effects of neurotensin applications were studied, on the one hand on the firing discharge of respiratory related neurons and on the other hand on the phrenic nerve activity and on arterial blood pressure. An increase of the firing frequency of respiratory related neurons was induced by neurotensin applied by iontophoresis or by pressure injection (0.005 33.5 fmol/s) on single neurons. In the latter case, neurotensin was active at concentration 10(3) times lower than glutamate. A bilateral apneustic pattern was induced on the phrenic nerve activities by microinjection of neurotensin (0.23 0.54 pmol/s) in one ventrolateral nucleus tractus solitarius without alteration of arterial blood pressure. These results suggest that the release of neurotensin in the nucleus tractus solitarius regulates respiratory rhythmogenesis by increasing the inspiratory duration. PMID- 3024775 TI - Effects of unilateral decortication on beta-adrenergic receptors in the remaining cortex and in the hypothalamus of female rats. AB - Many experiments have been performed to evaluate the physiological role of catecholaminergic mechanisms in gonadotropin release. The purpose of the present study was to determine the concentration of beta-adrenoreceptors in the remaining (right) cerebral cortex and in right and left hypothalamic halves of hemi decorticated female rats which exhibited elevated plasma gonadotropin levels as observed previously. The density of beta-receptors was measured using a high affinity beta-adrenergic ligand, iodocyanopindolol (ICYP). Scatchard estimates were obtained for maximum binding (Bmax fmol/mg of tissues) from pooled cerebral cortical and hypothalamic tissue of animals under several experimental conditions after hemi-decortication and sham operation. There was an increase in beta adrenoreceptor density in the remaining (right) cerebral cortex at all times examined in hemi-decorticate in comparison with the sham-operated animals (7 days, +10.9%; 21 days, +8.4%; 90 days, +22%; and 90 days plus ovariectomy, +34.8%). The number of beta-adrenoreceptors in the right hypothalamic half in hemi-decorticates decreased at 21 days (-42.20%) and then increased at 90 days (+76.63%) and 90 days plus ovariectomy (+51.75%) when compared with the left hypothalamic half. At the same time there were no significant changes in the sham operated animals when comparing the receptor density in the right and left hypothalamic halves, respectively. Thus, our results suggest a direct or indirect adrenergic pathway by which the left cortex can influence the right cortex and a crossed pathway to the contralateral hypothalamus changing adrenergic activity which can alter the beta-adrenergic receptor binding capacity in the hypothalamus. PMID- 3024776 TI - Vasoactive intestinal peptide stimulates cyclic AMP metabolism in choroid plexus epithelial cells. AB - Some peptides of the glucagon-secretin family were found to stimulate intracellular cyclic AMP accumulation in cultured bovine choroid plexus epithelial cells. Vasointestinal peptide and porcine intestinal peptide at concentrations of 30 and 300 nM, respectively, evoked 50-fold elevations of cyclic AMP; half-maximal responses were obtained with concentrations of 15 and 102 nM for the two peptides, respectively. Secretin and glucagon each produced 25 to 50-fold elevations of cyclic AMP at 330 microM, but showed no effect below 3 microM. Gastric inhibitory peptide and prealbumin had little or no response at any concentration tested. Experiments measuring the cellular cyclic AMP accumulation in response to pairs of peptides suggested that vasointestinal peptide, porcine intestinal peptide and secretin act through a common receptor. Studies with antagonists to isoproterenol and histamine indicated that this receptor is distinct from the beta-adrenergic and H2-histamine receptors known to exist on choroidal cells. PMID- 3024777 TI - Development of high affinity kainate binding sites in human and rat hippocampi. AB - Autoradiographic localization of kainate binding sites has been determined in developing human and rat hippocampi. The results suggest differences. In particular, a transient high density of sites occurs in the supragranular layer of the fascia dentata of the human hippocampus. PMID- 3024778 TI - Behavioural and biochemical evidence for a long-lasting decrease in GABAergic function elicited by chronic administration of FG 7142. AB - Chronic treatment with the beta-carboline derivative FG 7142 (15 mg/kg i.p. twice a day for 10 consecutive days) produced a long-lasting enhancement of shock induced suppression of drinking in rats, without affecting unpunished behaviour. This proconflict effect was observed up to 15 days after withdrawal from FG 7142. A significant sensitization to seizures induced by isoniazid, a drug known to inhibit GABAergic transmission, was also found to occur after long-term (25 days) withdrawal. Moreover, the density of low-affinity GABA receptors was decreased by 30% in the cerebral cortex of rats repeatedly injected with FG 7142 at 5 and 15 days after withdrawal. The capacity of high-affinity GABA receptors, as well as the apparent dissociation constants for both high- and low-affinity GABA receptors were unchanged. Similar modifications in [3H]GABA binding were also observed in the cerebellum. The enhancement of punishment suppressed behaviour, the sensitization to isoniazid-induced convulsions and the decrease in the density of low-affinity GABA receptors suggest that chronic administration of FG 7142 induces a persistent down-regulation of GABAergic transmission in the central nervous system. PMID- 3024779 TI - Characterization of benzodiazepine receptors in the bovine pineal gland: evidence for the presence of an atypical binding site. AB - Bovine and rat pineal benzodiazepine receptors were characterized using ligands with high affinities for either 'central-type' (CBR) or 'peripheral-type' (PBR) benzodiazepine receptors. The characteristics (Bmax = 83 +/- 10 fmol/mg protein, Kd = 3.88 +/- 0.46 nM) of benzodiazepine receptors in bovine pineal membranes measured with [3H]flunitrazepam (using flunitrazepam to define non-specific binding) were consistent with previously reported values. However, if non specific binding was defined using Ro 15-1788 (a selective CBR ligand), the Bmax and Kd of [3H]flunitrazepam decreased 51 and 58%, respectively. In addition, when using PK 11195 to determine non-specific binding, the Bmax of [3H]flunitrazepam binding to bovine pineal decreased further (approximately 80%, Kd decreased approximately 39%). Together, these observations strongly suggested the presence of PBR in the bovine pineal. Bovine pineal PBR characterized with [3H]PK 11195 revealed a high density (relative to CBR) of high affinity binding sites (Kd = 1.08 +/- 0.30, Bmax = 776 +/- 33.0 fmol/mg protein). In contrast, when [3H]Ro 5 4864 (1-20 nM) was used to define PBR, no binding was detectable. These observations are in sharp contrast to the rat pineal gland, in which both [3H]Ro 5-4864 and [3H]PK 11195 bind to a large number of PBR with high affinity (Kd approximately equal to 1.9 nM, Bmax approximately equal to 26 pmol/mg protein). Bovine pineal PBR were further characterized with compounds structurally related to either Ro 5-4864 or PK 11195.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3024780 TI - Characteristics of a Ca2+/calmodulin-dependent binding of the Ca2+ channel antagonist, nitrendipine, to a postsynaptic density fraction isolated from canine cerebral cortex. AB - Synaptic membrane (SM) and postsynaptic density (PSD) fractions isolated from the cerebral cortex (CTX) and cerebellum (CL) of the canine brain were found to contain one class of specific nitrendipine binding sites. The specific binding constants were: CTX-SM, Kd = 110 pM (Bmax = 126 fmol/mg protein); CTX-PSD, Kd = 207 pM (Bmax = 196 fmol/mg); CL-SM, Kd = 100 pM (Bmax = 65 fmol/mg); CL-PSD, Kd = 189 pM (Bmax = 80 fmol/mg). Treatment of the CTX-SM and CTX-PSD fractions with 0.5% deoxycholate and 1.0% N-lauroyl sarcosinate removed 88-91% and 42-51% of the nitrendipine binding, respectively, indicating that the major nitrendipine binding present in the SM fractions are of non-synaptic origin. Moreover, the percentages of total protein and specific nitrendipine binding removed from PSDs by these detergents were similar, indicating no preferential dissociation of the latter, and suggesting that the receptor protein is firmly bound and is probably an intrinsic component of the PSD fraction. Both Ca2+ and calmodulin were found to be important for the binding of nitrendipine to the CTX-SM and CTX-PSD fractions since: R24571, a calmodulin antagonist, was found to inhibit nitrendipine binding to the CTX-SM and CTX-PSD fractions with IC50 values of 1.1 microM and 0.9 microM, respectively; removal of Ca2+ from the CTX-SM and CTX-PSD fractions with 0.2 mM EGTA resulted in losses of specific nitrendipine binding of 80 and 90%, respectively; Ca2+ alone restored nitrendipine binding to EGTA pretreated CTX-SM fractions and not to CTX-PSD fractions, with the latter needing both Ca2+ and calmodulin to restore nitrendipine binding; EGTA treatment removed 14-16% and 89-91% of nitrendipine bound to the CTX-SM and CTX-PSD fractions, respectively, suggesting that calmodulin (but not Ca2+) is needed to maintain the nitrendipine-nitrendipine receptor-calmodulin complex; Ca2+-reconstituted EGTA pretreated CTX-SM fractions and the Ca2+ plus calmodulin-reconstituted EGTA pretreated CTX-SM and CTX-PSD fractions were found to have similar binding constants to those for the corresponding native, untreated fractions; and the Ca2+/calmodulin dependency on nitrendipine binding was similar to the well-known Ca2+/calmodulin dependency on phosphorylation in EGTA-pretreated PSD fractions. It needed much less Ca2+ to saturate Ca2+/calmodulin-dependent phosphorylation of the pretreated CTX-PSD fractions than the nitrendipine binding. Yet, less calmodulin was needed to saturate nitrendipine binding than the phosphorylation.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3024781 TI - [Silent infection by the hepatitis A virus]. PMID- 3024783 TI - Reversibility of ultrastructural, contractile function and Ca2+ transport changes in guinea pig hearts after global ischemia. AB - To understand the subcellular basis of contractile failure due to ischemia reperfusion injury, effects of 20, 60, and 90 min of global ischemia followed by 30 min of reperfusion were examined in isolated guinea pig hearts. Cardiac ultrastructure and function as well as Ca2+ transport abilities of both mitochondrial and microsomal fractions were determined in control, ischemic, and reperfused hearts. Hearts were unable to generate any contractile force after 20 min of ischemia and showed a 75% recovery upon reperfusion. However, there were no significant changes in the subcellular Ca2+ transport in the 20-min ischemic or reperfused hearts. When hearts were made ischemic for 60 and 90 min, the recovery of contractile force on reperfusion was 50 and 7%, respectively. There was a progressive decrease in mitochondrial and microsomal Ca2+ binding and uptake activities after 60 and 90 min of ischemia; these changes were evident at various times of incubation period and at different concentrations of Ca2+. Mitochondrial Ca2+ transport changes were only partially reversible upon reperfusion after 60 and 90 min of ischemia, whereas the microsomal Ca2+ binding, uptake and Ca2+ ATPase activities deteriorated further upon reperfusion of the 90 min ischemic hearts. Ultrastructural changes increased with the duration of the ischemic insult and reperfusion injury was extensive in the 90-min ischemic hearts. These data show that the lack of recovery of contractile function upon reperfusion after a prolonged ischemic insult was accompanied by defects in sarcoplasmic reticulum Ca2+ transporting properties and structural damage. PMID- 3024782 TI - Changes in interaction of bovine dentin phosphophoryn with calcium and hydroxyapatite by chemical modifications. AB - Dentin phosphophoryn was found to lose its precipitability with calcium ion when dephosphorylated or carboxylate-blocked, whereas precipitation of the intact protein was induced at calcium concentration as low as 2 mM. On the other hand, the adsorption capacity of phosphophoryn onto the hydroxyapatite surface increased when acidic groups were diminished by the above modifications. PMID- 3024784 TI - Effects of sulfhydryl reagents on nitroglycerin-induced relaxation of bovine coronary artery. AB - The mechanism whereby nitroglycerin relaxes vascular smooth muscle remains uncertain. A current hypothesis suggests that nitroglycerin reacts with critical cellular sulfhydryl groups to form an intermediate, which activates guanylate cyclase, resulting in cGMP accumulation and relaxation. This study investigated further the potential involvement of sulfhydryls in nitroglycerin-induced vascular smooth muscle relaxation by evaluating effects of a variety of sulfhydryl alkylating and reducing agents on responses to nitroglycerin and other relaxants in bovine coronary arterial strips submaximally contracted using 30 mM K. Whereas 10(-4) M 5,5'-dithiobis-(2-nitrobenzoic acid), 10(-5) MN ethylmaleimide, and 10(-4) MN-naphthylmaleimide did not affect nitroglycerin induced relaxation, 10(-4) MN-ethylmaleimide and 10(-4) M ethacrynic acid significantly inhibited relaxation induced by nitroglycerin. Both ethacrynic acid and N-ethylmaleimide at 10(-4) M also inhibited relaxation induced by sodium nitroprusside. N-ethylmaleimide, but not ethacrynic acid, inhibited relaxation induced by isoproterenol and forskolin. Ethacrynic acid significantly reduced both relaxation and cGMP elevation induced by both 10(-7) M nitroglycerin and 10( 7) M sodium nitroprusside. Ethacrynic acid, but not N-ethylmaleimide, significantly reduced relaxation induced by 8-Br-cGMP. Pretreatment with the sulfhydryl-containing agents N-acetylcysteine, 2-mercaptoethanol, or dithiothreitol, at 10(-3) M did not affect nitroglycerin-induced relaxation in nontolerant arteries. Similarly, N-acetylcysteine and dithiothreitol did not alter the depressed responses to nitroglycerin in arteries in which tolerance to nitroglycerin was induced in vitro. A slight but statistically significant reversal of nitroglycerin-tolerance occurred after treatment of tolerant arteries with 2-mercaptoethanol.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3024785 TI - The role of alpha-adrenergic receptors in carbon monoxide hypoxia. AB - The importance of alpha-adrenergic receptors in the cardiac output and peripheral circulatory responses to carbon monoxide (CO) hypoxia was studied in anesthetized dogs. Phenoxybenzamine (3 mg/kg i.v.) was injected to block alpha-receptor activity and the data obtained were then compared with those from a previous study of CO hypoxia in unblocked animals. Values for cardiac output, hindlimb blood flow, vascular resistance, and oxygen uptake were obtained prior to and at 30 and 60 min of CO hypoxia which reduced arterial oxygen content by approximately 50%. alpha-Adrenergic blockade resulted in a lower (p less than 0.05) control value for cardiac output than observed in unblocked animals, but no differences were present between the two groups at 30 or 60 min of CO hypoxia. Similarly, limb blood flow was lower (p less than 0.05) during the control period in the alpha-blocked group but rose to the same level as that in the unblocked animals at 60 min of COH. No change in limb blood flow occurred during CO hypoxia in the unblocked group. These findings demonstrated that during CO hypoxia alpha receptor mediated venoconstriction does not contribute to the cardiac output response and alpha-receptor mediated vasoconstriction probably does prevent a rise in hindlimb skeletal muscle blood flow. PMID- 3024786 TI - The inhibitory effects of some adenosine analogues on transmitter release at the mammalian neuromuscular junction. AB - An electrophysiological study was made of the effects of four adenosine analogues, 2-chloroadenosine (2-CIA), 5'-N-ethylcarboxamidoadenosine (NECA), L-N6 phenylisopropyladenosine (L-PIA), and 2-(p-methoxyphenyl)-adenosine (CV-1674) on neurotransmitter release in the mouse phrenic nerve - hemidiaphragm preparation. All four drugs decreased miniature end-plate potential frequency in a dose dependent manner. Evoked transmitter release in the cut diaphragm preparation was depressed by 2-CIA and CV-1674 to a similar extent. The ability of theophylline to antagonize the inhibitory effect of CV-1674 on spontaneous transmitter release was also established. On the basis of these results, the rank order of potencies was: L-PIA greater than NECA greater than 2-CIA greater than CV-1674. A clear classification of receptor type could not be made, since the ratio of potencies of L-PIA and NECA was narrow. Different slopes of the concentration-effect curves for 2-CIA and CV-1674 compared with L-PIA and NECA suggest an additional component to simple agonist action in their overall effects. PMID- 3024787 TI - L-649,923, sodium (beta S*, gamma R*)-4-(3-(4-acetyl-3-hydroxy-2-propylphenoxy) propylthio)- gamma-hydroxy-beta-methylbenzenebutanoate, a selective, orally active leukotriene receptor antagonist. AB - L-649,923, Sodium (beta S*, gamma R*)-4-(3-(4-acetyl-3-hydroxy-2 propylphenoxy)propylthio)- gamma- hydroxy-beta-methylbenzenebutanoate is a selective and competitive inhibitor of [3H]leukotriene D4 (Ki value of 400 nM) and to a lesser extent [3H]leukotriene C4 (Ki value of 8.6 microM) binding in guinea-pig lung homogenates. Functionally, it selectively antagonized contractions of guinea pig trachea induced by leukotriene C4, D4, E4, and F4 but not those induced by acetylcholine, histamine, serotonin, prostaglandin F2 alpha, or U-44069 (stable endoperoxide analogue). Schild plot analysis indicated a competitive inhibition of contractions of guinea-pig ileum induced by leukotriene D4 (pA2 8.1) and contractions of guinea-pig trachea induced by leukotrienes E4 and F4 (pA2 7.1 and 6.9, respectively). In contrast, contractions of guinea-pig trachea induced by leukotrienes C4 (pA2 7.2; slope 0.6) and D4 (pA2 7.2; slope 0.7) were inhibited in a noncompetitive fashion. In vivo, intravenously administered L-649,923 selectively blocked bronchoconstriction induced in anesthetized guinea pigs by leukotriene C4 and D4 (ED50 values i.v. 0.38 and 0.26 mg/kg, respectively) but not that induced by histamine, arachidonic acid, serotonin, U-44069, or acetylcholine. Following intraduodenal administration, L 649,923, blocked leukotriene D4 induced bronchoconstriction (5 and 10 mg/kg). The present findings indicate that selective antagonists, such as L-649,923, may be useful for defining the role of leukotrienes in diseases such as bronchial asthma. PMID- 3024788 TI - Ovine fetal adrenal maturation at term and during fetal ACTH administration: evidence that the modulating effect of cortisol may involve cAMP. AB - Exogenous ACTH1-24 promotes adrenal maturation in fetal sheep, and this effect appears to be modulated in part by cortisol (Challis et al. 1985). We have examined whether similar changes in adrenal metabolism of progesterone occur with ACTH-induced labour as at spontaneous term and whether the site of cortisol modulation is on adrenal steroidogenesis or at the level of cAMP generation. Chronically catheterized fetal sheep were infused in utero for 100 h between days 127 and 131 of pregnancy with P-ACTH, P-ACTH + metopirone, P-ACTH + metopirone + cortisol, or saline. After 100 h the metabolism of [3H]progesterone was measured in adrenal homogenates. Similar incubations were performed with adrenal tissue from fetal sheep at day 130 of pregnancy and at spontaneous labour. In the treatment groups of sheep, cAMP output by dispersed adrenal cells in response to ACTH added in vitro was also determined. Similar qualitative patterns of [3H]progesterone metabolism were found in adrenal homogenates after in vivo ACTH or at term. At both times there was an increase in cortisol and in total 17 alpha hydroxycorticosteroid accumulation and also evidence for increased activity of 11 beta-hydroxylase enzyme. The formation of total 17 alpha-hydroxycorticosteroids was not affected significantly by concurrent treatment in vivo with metopirone +/ cortisol. The accumulation of cAMP in vitro was increased after in vivo ACTH, attenuated after ACTH + metopirone, but statistical significance over controls was restored after ACTH + metopirone + cortisol treatment. We conclude that ACTH induced labour and spontaneous parturition in sheep is associated with qualitatively similar changes in progesterone metabolism by the fetal adrenal gland.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3024789 TI - Potentiation of salivary fluid secretion in ixodid ticks: a new receptor system for gamma-aminobutyric acid. AB - gamma-Aminobutyric acid (GABA), having minimal intrinsic activity, potentiates dopamine-induced fluid secretion in salivary glands of female ixodid ticks. Because the effect of GABA was similar to that of spiperone, we tested whether these two drugs act at a common recognition site. Potentiation was not augmented when salivary glands were exposed to supramaximal concentrations of spiperone (1 microM) plus GABA (100 microM). (+/-)-Sulpiride (100 microM), a spiperone antagonist in this system, also blocked GABA-induced potentiation. Picrotoxin (100 microM) and (-)-bicuculline (100 microM), two GABA antagonists, blocked GABA induced and spiperone-induced potentiation. Inhibition of GABA by picrotoxin and (-)-bicuculline was noncompetitive. Muscimol (an agonist at GABAA receptors) also potentiated dopamine-induced secretion. Baclofen (an agonist at GABAB receptors) did not elicit potentiation. We suggest that GABA may function as a neuromodulator for dopamine-induced fluid secretion in tick salivary glands. PMID- 3024790 TI - Vascular role of vasopressin in the presence and absence of influence from angiotensin II or alpha-adrenergic system. AB - The effects of a vasopressin (AVP) pressor antagonist, d(CH2)5Tyr(Me)AVP, on mean arterial pressure (MAP), total peripheral resistance (TPR), cardiac output (CO), and the distribution of CO were investigated by the microsphere technique in three groups of pentobarbital anesthetized rats: intact (I), saralasin pretreated (II), and phentolamine pretreated (III). Saralasin and phentolamine were infused intravenously to inactivate the renin-angiotensin and alpha-adrenergic systems, respectively. The AVP antagonist decreased MAP and TPR in all groups and it caused a greater depressor effect in groups II and III than in group I. In group I, AVP antagonist increased blood flow (BF) to the stomach and skin. In group II, AVP antagonist increased BF to the muscle and skin. In group III, AVP antagonist markedly increased BF to the muscle. Therefore, the degree of vasoconstrictor influence exerted by AVP in different vascular beds varies depending on endogenous vasomotor tone from the renin-angiotensin and (or) sympathetic nervous systems. PMID- 3024791 TI - Comparison of 1-oleoyl-2-acetyl-glycerol and phorbol myristate acetate as secretagogues for human neutrophils. AB - Cellular responses induced in human neutrophils by the synthetic diacylglycerol, 1-oleoyl-2-acetyl-glycerol (OAG), paralleled those induced by phorbol myristate acetate (PMA). Like PMA, OAG caused the preferential release of enzymes from specific granules and promoted superoxide (O2-) generation. The efficacy of OAG was similar to that for PMA, but its potency was lower by four orders of magnitude. First derivative kinetic analysis showed that rates of O2- generation elicited by PMA decayed exponentially in a first order manner; the half life was found to be 21 +/- 6 min. Results obtained in studies carried out with high OAG concentrations were similar except that after 40 min, the rate of decay increased and became complex order. This difference was attributed to the greater susceptibility of OAG to metabolic alteration, and was reflected in the NADPH oxidase activity of granule rich membrane fractions (GRF) of cells stimulated for 90 min with PMA or OAG. It was found that the O2- generating activity of the PMA treated GRF was significantly greater than that for the OAG treated fraction. Current evidence indicates that cellular responses arise from direct activation of protein kinase C by PMA-OAG. The stability of this complex and the bypassing of normal regulatory constraints may account for the relative longevity of the PMA-OAG O2- respiratory burst. PMID- 3024792 TI - Characterization of the stability functions and inverted repeat structure X3 of the IncFI plasmid ColV2-K94. AB - A region of the IncFI plasmid ColV2-K94 which showed homology to the sop partitioning genes of F was cloned and characterized in an attempt to study the stability functions of this element. The sop region contained the incD incompatibility determinant common to many IncFI plasmids, but could not confer on ColV2-K94 miniplasmids the same stable inheritance found in the intact ColV2 K94; thus, other functions appear to be required for efficient plasmid maintenance. Adjacent to the area of sop homology was the X3 region, which was found to contain three inverted IS1-like sequences. The X3 region of ColV2-K94 was similar in organization to the aerobactin iron uptake region of ColV3-K30, but ColV2-K94 lacked the ability to synthesize either the aerobactin siderophore or its outer membrane receptor. PMID- 3024793 TI - Speculations on the role of transmissible agents in the pathogenesis of Alzheimer's disease. AB - Unconventional agents and conventional viruses provide model systems to investigate the pathogenesis of Alzheimer's disease (AD). The essay which follows examines the hypothetical role of herpes simplex in AD and presents some generally applicable experimental approaches to detecting genes in brain tissues. The concluding section, on parallels between AD and diseases of the brain caused by unconventional viruses, defines strategies for isolating genes related to pathology. PMID- 3024794 TI - Lewy body dementia without Alzheimer changes. AB - Three patients with clinical Alzheimer's disease were found at postmortem examination to have Lewy-bodies and Lewy-like bodies in the cerebral cortex and the pigmented brainstem nuclei. Neuritic plaques were found in neocortical areas but no neurofibrillary tangles. The distribution of cortical neuronal inclusions correlated with the proposed projection of dopamine terminals. Neuronal cell loss was marked in the ventral tegmental area (paranigral nucleus) and the basal nucleus of Meynert, suggesting a defect in dopaminergic and cholinergic innervation of the cerebral cortex. Immunohistochemical investigations revealed positive staining of cortical Lewy- and Lewy-like bodies for monoclonal antibodies to phosphorylated neurofilaments (03-44, 06-17, 04-7). Also cerebral neurons containing no inclusions showed positivity, suggesting an early neurofilament abnormality, preceding the formation of Lewy-type inclusions. PMID- 3024795 TI - Further studies on the clinical features and clinicopathological findings of a syndrome of metabolic acidosis with minimal dehydration in neonatal calves. AB - A syndrome of metabolic acidosis of unknown etiology was diagnosed in twelve beef calves 7 to 31 days old. Principal clinical signs were unconsciousness or depression concomitant with weakness and ataxia. Other signs included weak or absent suckle and menace reflexes, succussable nontympanic fluid sounds in the anterior abdomen, and a slow, deep thoracic and abdominal pattern of respiration. The variation in clinical signs between calves was highly correlated (r = 0.87, P less than 0.001) with their acid-base (base deficit) status. Abnormal laboratory findings included reduced venous blood pH, pCO2 and bicarbonate ion concentration as well as hyperchloremia, elevated blood urea nitrogen, increased anion gap and neutrophilic leukocytosis with a left shift. Sodium bicarbonate solution administered intravenously effectively raised blood pH and improved demeanor, ambulation and appetite. All calves did well following a return to a normal acid base status. PMID- 3024796 TI - The alkalinizing effects of metabolizable bases in the healthy calf. AB - The alkalinizing effect of citrate, acetate, propionate, gluconate, L and DL lactate were compared in healthy neonatal calves. The calves were infused for a 3.5 hour period with 150 mmol/L solutions of the sodium salts of the various bases. Blood pH, base excess, and metabolite concentrations were measured and the responses compared with sodium bicarbonate and sodium chloride infusion. D gluconate and D-lactate had poor alkalinizing abilities and accumulated in blood during infusion suggesting that they are poorly metabolized by the calf. Acetate, L-lactate and propionate had alkalinizing effects similar to bicarbonate, although those of acetate had a slightly better alkalinizing effect than L lactate. Acetate was more effectively metabolized because blood acetate concentrations were lower than L-lactate concentrations. There was a tendency for a small improvement in metabolism of acetate and lactate with age. Sodium citrate infusion produced signs of hypocalcemia, presumably because it removed ionized calcium from the circulation. D-gluconate, D-lactate and citrate are unsuitable for use as alkalinizing agents in intravenous fluids. Propionate, acetate and L lactate are all good alkalinizing agents in healthy calves but will not be as effective in situations where tissue metabolism is impaired. PMID- 3024797 TI - Immunization of Canadian Armed Forces personnel with live types 4 and 7 adenovirus vaccines. PMID- 3024798 TI - DNA flow analysis of soft tissue tumors. AB - The cellular DNA content of 81 soft tissue tumors was determined by means of flow cytometry and related to conventional histologic classification of the same tumors. Comparison of histologic and cytometric analysis showed that all 23 benign tumors were diploid (normal DNA content), whereas the malignant group included both diploid and aneuploid (abnormal DNA content) lesions. There appeared to be a relationship between tumor grade and ploidy level in that 92% of Grade II, 28% of Grade III, and 11% of Grade IV lesions were diploid. Cell distribution analysis, feasible in 51 cases, disclosed that diploid lesions had a low proportion of S and G2 + M cells and most aneuploid lesions a high proportion, indicating a relationship between ploidy level and proliferative activity. The current study shows that solid mesenchymal tumors may be analyzed by DNA flow cytometry. Regardless of histogenetic type, it appears that benign and low-grade tumors are diploid and high-grade tumors, in general, are aneuploid. As to exceptions, DNA analysis may prove to give information beyond that obtained by subjective histologic interpretation. Thus, adequate follow-up might show that high-grade lesions with a diploid DNA content are associated with a better prognosis than expected from histologic classification. PMID- 3024799 TI - Cancer among Haitians in Florida. AB - Data on cancer rates from West Indian populations are scarce, and to the authors' knowledge there are no published data on cancer rates and distributions among Haitians. Proportional distributions of cancers among three groups of patients living in Florida were compared: Haitian born blacks, United States born blacks, and non-Haitian Caribbean born blacks. The incidence rate of cancer of the cervix among the Haitian and United States born black groups was also compared. Increased rates of certain malignancies associated with viral infection or immunodeficiency were found in the Haitian group. These tumors were hepatocellular carcinoma, nasopharyngeal carcinoma, reticulum cell sarcoma, Kaposi's sarcoma, and carcinoma of the uterine cervix. The age-adjusted incidence rate of carcinoma of the cervix was especially high among Haitian women even with a liberal estimate of the female Haitian population from whom the cases were drawn. Except for cancer of the cervix, the numbers of cancers of interest were small, and age-adjusted incidence rates were not calculated. Continued epidemiological study of larger numbers of patients is needed to evaluate these findings further. PMID- 3024800 TI - Human monoclonal antibodies derived from lymph nodes of a patient with breast carcinoma react with MuMTV polypeptides. AB - Nine human hybridoma cell lines were established from a fusion of axillary lymph node lymphocytes of a patient with breast ductal carcinoma with a human lymphoblastoid cell line. The human hybridoma nature of the fusion products was confirmed by chromosomal analysis and HLA typing. The hybridomas are stable over a year of growth, and can be frozen, thawed and regrown. The carcinoma cells of the patient harbor mouse mammary tumor virus (MuMTV) cross-reacting antigens. The patient's serum and the purified monoclonal antibodies reacted with MuMTV polypeptides. Radioimmunoprecipitation studies using labeled MuMTV confirmed the binding of the patient's serum to the viral proteins. None of the control immunoglobulins reacted with the virus. No binding of the hybridoma immunoglobulins was observed with two other retroviruses (avian myeloblastosis virus and simian sarcoma virus). The ligand binding characteristics of the monoclonal antibodies suggest binding to epitopes on the various structural virus polypeptides. These monoclonal antibodies may serve as a probe to analyze the MuMTV-human breast carcinoma relationship. PMID- 3024801 TI - Small cell carcinoma of the lung. A progress report of 15 years' experience. AB - To assess the results of therapeutic advances in the treatment of small cell carcinoma of the lung (SCCL) achieved during the past 15-year period at a single large institution, 508 patients treated between 1968 and 1982 were divided into two groups: 157 patients (66 in the category of limited-stage disease and 91 in the extensive-stage disease category) treated with low-dose small-volume radiotherapy (RT) (time dose fractionation [TDF] 49-66) and with cyclophosphamide alone or a COPP program during the first period of 7 years (1968-1974); 351 patients (180 in limited and 171 in extensive stage) treated with multidrug chemotherapy (CT) and high-dose large-volume RT (TDF 73-89) during the second period of 8 years (1975-1982). For patients with limited-stage cancer, 5-year actuarial survivals were 3% versus 7% for the periods 1968-1974 versus 1975-1982, respectively, P less than 0.01. For patients with extensive-stage cancer, the median survival time (MST) and 2-year actuarial survivals were 5 months and 2% versus 7 months and 4% for the periods 1968-1974 versus 1975-1982, respectively. To evaluate the outcome of a contemporary approach, i.e., CT alone, with RT reserved for locoregional failure, 180 patients with limited-stage cancer who were treated (1975-1982) were further analyzed for MST, 2- and 5-year actuarial survival figures, and local-tumor control rates according to the therapeutic approaches employed: CT + RT (112); CT alone (36); RT alone (17); and surgery (S) +/- CT +/- RT (15). Although the 36 patients in CT alone seems a small number, 17 of the 36 patients were enrolled in this approach in 1981-1982, reflecting a shift of emphasis from RT to CT. The MST and 2-year actuarial survival figures were 11 months and 0% versus 13 months and 21% for CT alone versus CT + RT respectively, P less than 0.05. CT + RT achieved a 5-year cure rate of 8%. S +/- CT +/- RT or RT alone also achieved 5-year cure rates of 8% and 10.5%, respectively, in selected subsets of patients. Local relapse rates were 80% (29/36) versus 25% (28/112) for CT alone versus CT + RT. These data emphasize the importance of thoracic RT given at the early phase of treatment to improve long term survival for patients with limited-stage SCCL. PMID- 3024802 TI - Hemophagocytic syndrome complicating T-cell acute lymphoblastic leukemia with a novel t(11;14)(p15;q11) chromosome translocation. AB - A case of hemophagocytic syndrome that developed in a patient with T-cell acute lymphoblastic leukemia (ALL) with a novel chromosome translocation involving 14q11 is reported. A 15-year-old boy with T-cell ALL in relapse showed leukemic cells with an abnormal karyotype of 46,XY,-15,t(11;14)(p15;q11), +der(15)t(15;?)(p11;?). Pancytopenia and extensive hemophagocytosis by macrophages in the bone marrow were observed after reinduction chemotherapy and again at the terminal stage. At autopsy, infiltration of such cells was also found in other organs. The findings suggested occurrence of hemophagocytic syndrome probably associated with cytomegalovirus (CMV) infection. The t(11;14)(p15;q11) may be a novel translocation specific for T-cell ALL, and conceivably, the association of T-cell ALL with the histiocytosis in this patient may not have been coincidental. PMID- 3024803 TI - A survey of human cancers for human papillomavirus DNA by filter hybridization. AB - We examined 217 tissue samples of various human malignancies for the presence of human papillomavirus (HPV) DNA using low-stringency filter hybridization techniques. These techniques were sufficiently sensitive for crosshybridization of the HPV DNA probes to all the known types of papillomavirus DNAs, both human and animal. Approximately 2% of the cancers analyzed contained HPV DNA. These included carcinomas of the lung, cecum, tongue, and neck. Three of four cancers contained HPV-16-related nucleotide sequences. Thus, in addition to previous data demonstrating the association of HPV DNA with certain cancers of the skin and genital tract, data is presented that indicates that several additional human cancers also contain HPV-related nucleotide sequences. PMID- 3024804 TI - Medial regression and its functional significance in tumor-supplying host arteries. A morphometric study of hepatic arteries in human livers with hepatocellular carcinoma. AB - Livers from 30 patients with hepatocellular carcinoma were submitted to morphometric analysis of the arterial smooth muscles to correlate the peculiar circulation in cancer, i.e., the lack of flow regulation, with the structure of blood vessels in and around the tumors. The anatomical radius and medial thickness of the cross-sectioned arteries were determined in a standardized state of the circularly stretched, internal elastic membrane. It was shown that not only the medial smooth muscles were extremely hypoplastic in arterioles contained in the carcinoma, but the media of tumor-supplying host arteries was also significantly thinner than in the controls, even in segments distant from the tumor, showing more or less lowered regulatory activity of the whole tumor bearing arterial tree. Under these circumstances, dissection of the host arteries was frequently found to arise, evolving into the typically non-muscularized tumor vessels. The authors consider the abnormal hemodynamics of cancer to be fully explainable from these vascular alterations. PMID- 3024805 TI - Melanocytic neuroendocrine carcinoma of the thymus. AB - A neuroendocrine carcinoma of the thymus with an ectopic adrenocorticotropic hormone (ACTH) syndrome and melanocytic differentiation is described. ACTH, neuron-specific enolase (NSE) and S-100 protein were identified in the tumor by immunocytochemistry. Neurosecretory granules and melanosomes could be demonstrated in different cell populations by electronmicroscopy. The clinicopathologic findings are presented. The literature is briefly discussed. PMID- 3024807 TI - Orchidectomy alone in stage I nonseminomatous testicular germ cell tumors. AB - Fifty-four patients with Stage I nonseminomatous testicular germ cell tumors (NSTGCT) were treated from 1982 to 1984. In 1982 and 1983, the orchidectomy was followed by an exploratory laparotomy to conclude the dissemination study. In 1984, laparotomy was performed only if indicated. The mean follow-up was 29 months. A relapse occurred in 11 patients (20%). The relapse rate in patients who underwent exploratory laparotomy was as high as that in patients who did not. All patients treated for relapse by chemotherapy and surgery entered a complete remission for at least 1 year. It proved impossible to establish criteria for prediction of a subsequent relapse. Both serum tumor marker assays and roentgenography are important aids in diagnosing a relapse. With careful follow up of Stage I NSTGCT patients, a wait-and-see attitude can be adopted until a relapse occurs. PMID- 3024806 TI - The significance of atypia within teratomatous metastases after chemotherapy for malignant germ cell tumors. AB - The completely resected teratomatous metastases of 55 patients who had been treated with cisplatin-based combination chemotherapy for non-seminomatous germ cell tumors were reviewed to see if cellular atypia had an effect with respect to recurrent disease. The degree of atypia of the epithelial and mesenchymal elements was assessed on the basis of the cytologic features and mitotic activity. Twenty-three percent of the cases contained high-grade epithelial elements, whereas high-grade mesenchymal elements occurred in 18% of the cases; in addition there were nine cases classified as showing frankly malignant teratomatous elements. The presence of cytologically disturbing epithelial and mesenchymal elements (which, however, lacked an invasive malignant pattern) correlated with an increased incidence of recurrent teratoma compared to less atypical teratomatous elements (23% vs. 6% for epithelial elements, and 18% vs. 9% for mesenchymal elements, respectively). This difference, however, was not statistically significant (P greater than 0.05). There was no correlation between teratomatous atypia and recurrent, non-teratomatous germ cell tumor. The presence of an invasive malignant pattern did identify patients at significantly increased risk for recurrent teratoma-derived tumor. The authors conclude that cytologic atypia in the absence of invasion is not sufficient justification for altering the usual therapeutic strategies for patients with teratomatous metastases. PMID- 3024808 TI - Mucinous colon carcinoma in a black family. AB - Carcinoma of the large bowel developed in an autosomal dominant pattern in 13 members of a black-American family. Seven members were affected prior to initial ascertainment of the family in 1976. Thereafter, the remaining six were affected while 0.2 cases were expected (p less than 0.001). Median age at diagnosis of colon cancer was 39 years (range, 22-62 years) in this family, compared with 65 years among black-Americans, in general. Histologic review of surgical specimens from six patients and medical record data for a seventh patient showed mucinous adenocarcinoma of the colon, an uncommon histologic variant. Studies of several family members a decade ago had revealed no biologic markers of cancer susceptibility. PMID- 3024809 TI - Cloning of a non-c-myc DNA fragment from the double minutes of a human colon carcinoid cell line. AB - The cell line COLO 320 DM, derived from an untreated human colon carcinoid tumor, was subcloned to obtain a population (Cl 11) with an average of 37 double minutes (DM) per cell. Fractionation of the chromosomes by differential centrifugation yielded a fraction enriched in DM. DNA isolated from the DM-enriched fraction was inserted into the Pst I site of pBR322. One clone, p446, representative of a number of similar clones, contained a region complementary to genomic unique sequences (region p446U). Southern blot analysis using COLO 320 DNA, and DNA from two other cell lines derived from the same biopsy, COLO 320 HSR and COLO 321 HSR, demonstrated amplification and rearrangement of sequences complementary to p446U when compared with 28 different tumor and normal cell lines, some of which contained DM or homogeneously staining regions (HSR). COLO 320 DM Cl 11 had approximately 110 copies per cell of the p446U sequence, or three copies per DM. COLO 320 HSR, which contained one HSR, had 35 copies per cell, while COLO 321 HSR, which contained two HSR, had 700 copies. In addition, p446U did not hybridize with insert sequences of recombinant plasmid pHM(E + H), which includes the human c-myc coding region, 3 kb of upstream flanking sequences and 0.5 kb of downstream flanking sequences, or with an exon 3 probe, pMYC RI-CLA. Amplification of p446U was also not seen in cell lines containing amplified c-myc or N-myc genes. These results indicate that more than one sequence may be amplified in DM or HSR containing tumor cells, but that they need not be amplified together in other tumors. PMID- 3024810 TI - Cytogenetic analysis in human breast carcinoma. II. Seven cases in the triploid/tetraploid range investigated using direct preparations. AB - This report presents karyotypes of seven breast carcinomas with high ploidy from our total of 111 cases. These karyotypes were highly complex and there was no indication of a specific deletion of 16p12----pter as indicated by the previous analysis of some near-diploid tumors. A comparison of numerical changes did not demonstrate a common loss of chromosome #16 as in the near-diploid tumors, but an equivalent loss of chromosomes #8 and #13 was found. PMID- 3024811 TI - Primitive neuroectodermal tumor cell lines: chromosomal analysis of five cases. AB - A cytogenetic analysis of primitive neuroectodermal tumor (PNET) cell lines was undertaken. PNET are presumed to be embryologically related to, but clinically and histologically distinct from, other tumors of neuroectodermal origin, including neuroblastoma and retinoblastoma. No single chromosome abnormality was found in all five of the tumors studied. In three of the five cases, however, additional 1q material [either as extra chromosome #1 or i(1q)] was found in all cells, and in two of the five, monosomy 13 was noted in all cells; the possible significance of these findings is discussed. PMID- 3024812 TI - Aberrations of chromosome 8 in mixed salivary gland tumors--cytogenetic findings on seven cases. AB - Cytogenetic findings on seven mixed salivary gland tumors are reported herein. The involvement of chromosome #8 in clonal chromosome aberrations in five of the seven tumors was particularly noteworthy. Four tumors had translocations involving chromosome #8 and one or two other chromosomes (#3, #7, #9, #13). The fifth showed a deletion of parts of the long arm of chromosome #8. In an attempt to define the critical segment on chromosome #8, we have identified the part between 8q11 and 8q13 as the critical region involved in all rearrangements. Thus far, our results confirm the results of the Swedish group, though the percentage of cases having #8 abnormalities is somewhat higher in our small series. The relationship between the two groups of cases, those with and those without chromosome abnormalities, will be discussed. PMID- 3024813 TI - Identification of marker chromosomes by restriction endonucleases/Giemsa technique in neoplastic cells. AB - The most recent addition to the various selective staining methods is the technique using restriction endonucleases to digest the metaphase chromosomes and subsequent staining by Giemsa (endonuclease/Giemsa technique). One of the endonucleases, AluI, which induces a characteristic modified C-band pattern is evaluated for its application in cancer cytogenetics using malignant lymphoid cells. The pattern obtained by AluI/Giemsa has been routinely useful in identifying some of the unusual markers that are difficult by routine banding. The centromeric regions are found to be far more heterogeneous by AluI resulting in frequent heteromorphic markers on several chromosomes. This has provided additional information to detect the donor cells in grafts after bone marrow transplantation. PMID- 3024814 TI - Neuroectoderm-associated antigens on Ewing's sarcoma cell lines. AB - The histogenesis of Ewing's sarcoma, the second most frequent primary bone tumor in humans, remains controversial. Ten Ewing cell lines were analyzed by immunological methods. Surface antigens recognized on Ewing cells were found to be related to the neuroectoderm lineage. They included ganglioside GD2, a marker of neuroectodermal tissues and tumors, and an acidic glycolipid detected by monoclonal antibody HNK-1 in the nervous system. The P61 rat monoclonal antibody that reacts with a peptide moiety of neural cell adhesion molecule (N-CAM) and a rabbit antiserum raised to purified mouse N-CAM also stained Ewing cells. Flow cytometry analysis performed using these reagents allowed the definition of four distinct Ewing phenotypes: all reagents equally stained group 1 lines; group 2 lines were strongly reactive with anti-N-CAM reagents, by contrast with a fainter staining with HNK-1 and anti-GD2 antibodies; all reagents but P61 were strongly reactive with group 3 lines; in group 4, Ewing lines were stained by P61 but only poorly by the anti-N-CAM antiserum. Several antibodies to melanoma and neuroblastoma associated antigens including two monoclonal antibodies to the nerve growth factor receptor were also found to react with Ewing cells. By contrast, all antibodies detecting antigens specifically expressed in hematopoietic cell lineages were totally unreactive. HLA class II antigens were never detected while the level of expression of class I antigens varied to a large extent. Ewing cells are characterized by a specific t(11;22)(q23-24;q12) translocation also observed in neuroepithelioma, a neuroectodermal tumor. Thus, Ewing's sarcoma cells share antigenic and karyotypic features with derivatives of the neuroectoderm possibly indicating a related histogenesis. PMID- 3024815 TI - Cell cycle dependence of sister chromatid exchange induction by DNA topoisomerase II inhibitors in Chinese hamster V79 cells. AB - The cell cycle dependence of sister chromatid exchange (SCE) induced by topoisomerase II inhibitors was studied in Chinese hamster V79 cells. 4'-(9 Acridinylamino)methansulfon-m-anisidide, which increases the concentration of covalently linked DNA-topoisomerase II complexes (cleavable complexes), induces SCE strongly in only a short period of the cell cycle. The sensitive period was identified an occurring in early to mid-S phase through the use of labeled thymidine incorporation and flow cytometry. Novobiocin, an inhibitor which prevents formation of the cleavable complex, did not induce SCEs in any part of the cell cycle. However, novobiocin did decrease the level of 4'-(9 acridinylamino)methansulfon-m-anisidide-induced SCEs. These results indicate that the cleavable complex may be important in 4'-(9-acridinylamino)methansulfon-m anisidide-induced SCE. PMID- 3024816 TI - Suppression of in vivo growth of human cancer solid tumor xenografts by 1,25 dihydroxyvitamin D3. AB - It has been demonstrated previously that several human cancer cell lines possess specific, high affinity receptors for 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3, calcitriol] and that 1,25-(OH)2D3 and certain of its metabolites inhibit the growth in vitro of several human breast cancer and malignant melanoma cell lines, i.e., analogous to the effect of estrogens on breast cancer. Furthermore, it has been shown that 1,25-(OH)2D3 and one of its synthetic analogues prolonged the survival in mouse leukemia, induced by inoculation of leukemic cells into syngeneic mice. However, until now no growth-inhibitory effect of 1,25-(OH)2D3 has been demonstrated in vivo for human cancer cells or for solid cancers. This paper describes the suppression by 1,25-(OH)2D3 of the growth of human cancer cell-derived xenografts in immune-suppressed mice. However, the 24-hydroxylated metabolite and the 24-difluorinated analogue of 1,25-(OH)2D3, both of which are active in vitro, were ineffective in this xenograft model system. The suppression by 1,25-(OH)2D3, which was achieved without significant toxicity, was observed with xenografts derived from two 1,25-(OH)2D3 receptor-positive cell lines (COLO 206F, derived from a colonic cancer, and COLO 239F from a malignant melanoma) but not in those from a receptor-negative line (RPMI 7932, also derived from a malignant melanoma). These studies demonstrate that pharmacological doses of 1,25 (OH)2D3 can be tolerated in the presence of a low calcium diet and that these doses can suppress the growth of human solid xenograft tumors in vivo. This is the first report of 1,25-(OH)2D3 growth suppression of solid tumors derived from human cancer cells in an in vivo model system, and it supports the hypothesized potential of the hormone in the treatment of 1,25-(OH)2D3 receptor-positive human cancers. PMID- 3024817 TI - Effect of cell proliferation and chromatin conformation on intercalator-induced, protein-associated DNA cleavage in human brain tumor cells and human fibroblasts. AB - Antineoplastic intercalating agents such as 4'-(9-acridinylamino)methanesulfon-m anisidide (m-AMSA) stabilize a cleavable complex between topoisomerase II and DNA. The production of protein-associated DNA cleavage in whole cells exposed to m-AMSA is thought to represent the cellular correlate of this topoisomerase II mediated reaction. Protein-associated DNA cleavage can be quantified in mammalian cells by using alkaline elution technology. In an attempt to understand the impact of phenotypic and biochemical cellular characteristics on protein associated DNA cleavage, we quantified m-AMSA-induced DNA cleavage in quiescent or proliferative normal human fibroblasts (cell strain 1508) and human glioblastoma cells (line T98G) as well as in asynchronously proliferating HeLa cells. The magnitude of DNA cleavage in quiescent fibroblasts and quiescent glioblastoma cells was identical and low relative to that observed in the HeLa cells. The magnitude of DNA cleavage was enhanced in both cell types following proliferation. This enhancement was greater in the glioblastoma cells than in the fibroblasts. These results were not due to alterations in cellular m-AMSA uptake. Chromatin was more elongated (open) in the quiescent glioblastoma cells than in the quiescent fibroblasts (as visualized by using the premature chromosome condensation assay), suggesting chromatin accessibility to drug per se may not be a critical determinant of the magnitude of m-AMSA-induced DNA cleavage. The onset of the enhanced m-AMSA-induced DNA cleavability that accompanied proliferation closely followed the formation of regions of localized chromatin decondensation, a late G1 event, and coincided with the onset of enhanced thymidine uptake, a marker for the onset of S phase. m-AMSA-induced cytotoxicity was also enhanced in proliferating compared with quiescent cells. The major finding of this study is that the cellular target for m-AMSA, putatively topoisomerase II, is more susceptible to drug action in proliferating cells than in quiescent cells. Effects of chromatin conformation or cellular phenotype upon topoisomerase II mediated events such as m-AMSA-induced DNA cleavage are less certain. PMID- 3024818 TI - Relationship between the neurotoxicity of the hypoxic cell radiosensitizer SR 2508 and the pharmacokinetic profile. AB - Complete pharmacological data from 71 patients treated on the phase I trial of SR 2508 were analyzed to see if the dose-limiting toxicity of peripheral neuropathy is related to the individual patient's pharmacokinetic profile. In a retrospective analysis, the risk of toxicity was best predicted by using the bivariate model of total drug exposure and the time over which the treatment course was given. Drug exposure [area under the curve (AUC)] for a single treatment was calculated by the integral over time of the serum concentration of SR 2508. Since the AUC was constant during the course of a patient's treatment, the total drug exposure (total-AUC) was estimated by the product of the AUC times the number of drug administrations. While the clinical efficacy of hypoxic cell sensitizers remains to be proven, SR 2508 is better tolerated than its predecessors, misonidazole and desmethylmisonidazole, as three times the amount of SR 2508 can be given. If this model is confirmed in the current phase II and III trials, the probability of developing neuropathy would be predictable for an individual patient from measurements made at the time of the first drug dose, allowing for the adjustment of drug schedule to avoid all but minor toxicity. PMID- 3024819 TI - Analysis of murine cellular receptors for tumor-killing factor. AB - Receptors for tumor-killing factor (TKF) on the surface of murine cells were analyzed using radioiodinated TKF. Not only sensitive cells but also insensitive cells were found to have specific receptors. Among the sensitive cells, no clear relation was observed between the number of receptors on the cell surface and sensitivity to TKF. Compounds affecting microfilaments (cytochalasin B and D) and microtubules (colchicine and Colcemid) significantly inhibited cytolysis of sensitive cells induced by receptor-bound TKF. It is concluded that internalization of receptor-bound TKF is a prerequisite for triggering cytolysis. PMID- 3024820 TI - Differential modulation of human chorionic gonadotropin production by methotrexate in normal and malignant placental cultures and its increase by dibutyryl cyclic adenosine monophosphate and/or actinomycin D in normal cultures. AB - The influence of methotrexate (MTX), dibutyryl cyclic AMP, and actinomycin D on production of human chorionic gonadotropin (HCG) in normal first trimester human placental organ cultures was compared. Actinomycin D (10(-8) to 10(-6) M) elevated HCG production by as much as 3.5-fold in normal placenta, and a 2-fold increase in HCG levels was obtained by treatment with dibutyryl cyclic AMP (1 mM) and theophylline (1 mM). The combination of dibutyryl cyclic AMP (1 mM) plus theophylline (1 mM) and actinomycin D (10(-8) M) additively enhanced HCG production by 4.5-fold. In contrast, HCG levels in normal placental organ cultures were unaffected by MTX (10(-8) to 10(-5) M) despite several differing treatment regimens. The JAr line of human choriocarcinoma cells, on the other hand, exhibited an 8-fold increase in HCG levels following MTX exposure (10(-7) M). Incorporation of selected radiolabeled precursors of the de novo and salvage pathways of DNA synthesis was evaluated to assess potential metabolic alterations underlying the differential HCG response of these cultures to MTX. Deoxyuridine incorporation into DNA was decreased similarly in both normal and malignant placenta following MTX exposure. However, deoxycytidine incorporation was inhibited by MTX in normal placental cultures but was elevated by as much as 4 fold in JAr cultures exposed to MTX. Thymidine incorporation into DNA was increased in both groups in the presence of MTX; however, thymidine incorporation was more profoundly stimulated (5-fold) in normal placenta than in JAr cultures (2.5-fold). These data indicate dissimilar utilization of the de novo and salvage pathways of DNA synthesis by these cultures which may explain their differential responsiveness to MTX. PMID- 3024821 TI - Hepatic drug-metabolizing enzymes in primary and secondary tumors of human liver. AB - Significant increases in activities of epoxide hydrolase, UDP glucuronosyltransferase, and glutathione S-transferase, and marked reductions in cytochrome P-450 mixed-function oxidase systems occur in hyperplastic nodules induced in rat liver by chemical mutagens. In contrast, activities of both oxidative (Phase I) and conjugative (Phase II) enzymes are decreased in hepatocellular carcinomas induced by peroxisome proliferators. The present work compares alterations induced by chemical mutagens or peroxisome proliferators with changes in enzyme activities that occur in primary and secondary hepatic tumors in man. The above activities, along with beta-glucuronidase and arylsulfatase, were measured in liver samples from 6 normal livers obtained at immediate autopsy, and liver specimens obtained by surgical biopsy from the following patients: 8 with hepatomas, 5 with nonmetastatic colorectal carcinomas, and 14 with metastatic colorectal carcinomas. Cytochromes P-450MP and P-450NF in addition to epoxide hydrolase were measured by immunoquantitation. Enzymes involved in conjugation reactions were either assayed fluorometrically (UDP glucuronosyltransferase, beta-glucuronidase, sulfotransferase, and sulfatase) or spectrophotometrically (glutathione S-transferase) using umbelliferyl substrates or 1-chloro-2,4-dinitrobenzene. Secondary hepatic tumors showed no significant change in drug-metabolizing enzymes, in contrast to primary hepatomas, which displayed decreases in all of the measured drug metabolizing enzymes. Arylsulfatase was markedly depressed in primary hepatomas (14% of normal values). Thus, activities of drug-metabolizing enzymes in human primary tumors resemble those associated with altered hepatic foci induced by peroxisome proliferators such as ciprofibrate. The marked decreases in sulfatase that occurred in primary but not in secondary human tumors suggest that sulfation of endogenous compounds and xenobiotics may differ in patients with primary and secondary hepatic tumors. PMID- 3024822 TI - Somatostatin receptors in human endocrine tumors. AB - A selection of 90 mainly endocrine but nonpituitary tumors have been tested for their content of specific somatostatin receptors using receptor autoradiography. Somatostatin receptors were detected in the following tumors: in all 5 meningiomas tested; in 3 of 39 malignant breast tumors; and in 3 growth hormone releasing factor-producing tumors, i.e., one mediastinal carcinoid, one intestinal carcinoma, and its liver metastasis. Receptor density varied greatly among individual tumors. Some of the positive tumors were biochemically characterized using in vitro binding assay and were shown to have saturable and high affinity receptors with pharmacological specificity for somatostatin. The following tumors did not contain somatostatin receptors: prostate carcinomas (n = 17); prostate hyperplasia (n = 2); ovarian carcinomas (n = 6); endometrial carcinomas (n = 4); primary liver cell carcinomas (n = 3); pheochromocytomas (n = 3); aldosteronomas (n = 2); medullary thyroid carcinomas (n = 2); one adrenocorticotropic hormone-secreting pulmonary carcinoid; one astrocytoma; one neurofibroma; one lung tumor; and one bladder tumor. Somatostatin receptors can be found in benign or malignant tumors, originating in part from tissue not primarily known as a somatostatin target. The biological function of such receptors is, therefore, partly unknown. If they can mediate antiproliferative properties, as has been suggested to be the case for somatostatin receptors in selected endocrine tumors in rats and humans, the present data could be of potential therapeutic interest. PMID- 3024823 TI - Metabolic epidemiology of colon cancer: effect of dietary fiber on fecal mutagens and bile acids in healthy subjects. AB - Because of potential significance of fecal mutagens and secondary bile acids in the pathogenesis of colonic cancer and of inverse association between dietary fiber and colonic cancer risk, the effect of dietary wheat and rye fiber on fecal mutagenic activity and bile acid levels was studied in 15 healthy men and women who were consuming high fat/moderately low fiber diets and excreting high levels of fecal mutagens and bile acids. Each participant provided two 24-h stool specimens and a 3-day diet record while consuming their normal diet (control). All subjects were then asked to consume their normal diet plus 11 g of supplemental fiber per day in the form of whole grain bread for 4 weeks. During the last week of diet intervention, each subject provided two 24-h stool specimens and a 3-day dietary record. Fecal samples collected from both periods were analyzed for bile acids and for mutagens using Salmonella typhimurium strains TA98 and TA100 with and without microsomal activation. The concentration of fecal secondary bile acids was significantly lower during the fiber supplemental period in all subjects. Fiber supplementation also inhibited the fecal mutagenic activity in TA100 and TA98 with and without microsomal activation. Thus, the increased fiber intake in the form of whole wheat and rye bread may reduce the production and/or excretion of fecal mutagens and decrease the concentration of fecal secondary bile acids in humans. PMID- 3024825 TI - Re: A case-control study of hepatocellular carcinoma and the hepatitis B virus, cigarette smoking, and alcohol consumption. PMID- 3024824 TI - Detection of human papillomavirus DNA in invasive carcinomas of the cervix by in situ hybridization. AB - An examination of 27 invasive cancers of the cervix was performed using the technique of in situ hybridization using human papillomavirus DNA probes. Four tissues, previously found to harbor papillomavirus DNA by filter hybridization, were confirmed by in situ analysis. One further tissue never previously studied was also found to be positive by in situ hybridization. Overall, we found 33% of invasive cancers of the cervix to contain human papillomavirus DNA. In contrast, 55% of carcinoma in situ and severe dysplasia of the cervix were found to be positive for human papillomavirus DNA. These results confirmed that the sample population of patients in our studies have a relatively low association of human papillomavirus DNA with invasive cancers of the cervix and that in situ hybridization provides an effective complementation to filter hybridization for human papillomavirus-infected tumors. PMID- 3024827 TI - Phase II trial of mitomycin, vindesine, and hexamethylmelamine in metastatic non small cell bronchogenic carcinoma. AB - Mitomycin (10 mg/m2 iv on Day 1), vindesine (3 mg/m2 iv on Days 1 and 8), and hexamethylmelamine (100 mg/m2/day orally on Days 1-14) was administered to 32 patients with metastatic non-small cell bronchogenic carcinoma. No patient had been previously treated with chemotherapy and Eastern Cooperative Oncology Group (ECOG) performance status was 0-1 in 21 of 32 patients. Eleven partial responses (34%) were observed, with a median duration of 9 weeks. No complete responses were observed in this group of patients, whose median survival duration was 22 weeks. Moderate leukopenia (median leukocyte count nadir, 2500/mm3) was the major toxic effect. Although this regimen is active and relatively nontoxic, it will not be utilized in future ECOG trials because it has not produced an apparent improvement in survival duration. PMID- 3024826 TI - Combined alkylators and multiple-site irradiation for extensive small cell lung cancer: a Southwest Oncology Group Study. AB - The Southwest Oncology Group performed a study for patients with extensive small cell lung cancer in which two hypotheses were tested: that combined alkylating agents for induction might improve the initial response rate; and that multiple site, involved-field consolidation with irradiation in patients with response to chemotherapy might further improve response quality and duration. A regimen of combined alkylating agents plus vincristine [carmustine, thiotepa, vincristine, and cyclophosphamide (BTOC)] was compared to our standard program of cyclophosphamide, doxorubicin, and vincristine (CAV). Patients with bone marrow metastases and/or disseminated bone metastases were randomized to BTOC or CAV as initial induction therapy, but received no multiple-field irradiation afterward. Among 189 such patients, the overall response rates were similar (48% to BTOC and 61% to CAV, with complete remission in 5% and 15%, respectively). Toxic effects were also comparative, with 23% sustaining life-threatening or fatal complications on BTOC and 16% on CAV. Median survival (5.9 and 7 months for BTOC and CAV, respectively) and overall survival were not different, and are superimposable upon our previously reported results with CAV alone. For patients with no evidence of bone marrow involvement and no metastases identified beyond the confines of ipsilateral hemithorax, liver, and brain, the plan of therapy consisted of induction chemotherapy (BTOC or CAV) every 3 weeks for three cycles, followed by multiple-field irradiation consolidation to sites of prior involvement. Among 239 such patients, response rates to therapy initiated with BTOC and CAV were also similar: 54% for BTOC and 62% for CAV, with complete response in 16% and 13%, respectively. However, the program of BTOC followed by multiple-site irradiation was associated with a higher incidence of severe or life-threatening thrombocytopenia (23% versus 8% for CAV), accounted for almost entirely by patients receiving hepatic irradiation after chemotherapy, among whom the incidence of this complication (grade 3 or 4) was 76% and 14%, respectively. Other toxic effects, including granulocytopenia, were comparable. With the exception of thrombocytopenia in the cited subgroup, multiple-field irradiation consolidation proved feasible and tolerable, with improved quality of response evident after its initiation in 24% of 135 patients. However, a similar proportion developed disease progression during or shortly after radiation therapy, and the median survival of patients who received irradiation was only 9 months (7 months from the start of consolidation).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3024828 TI - Addition of etoposide to cyclophosphamide, doxorubicin, and vincristine for remission induction and survival in patients with small cell lung cancer. AB - A total of 116 patients with small cell lung cancer were randomized to receive either: cyclophosphamide, 750 mg/m2, doxorubicin, 50 mg/m2, and vincristine, 2 mg iv (Regimen A), or the same drugs plus etoposide, 100 mg/m2 iv daily for 2 days (Regimen B) every 3 weeks. Complete responders received whole-brain radiation therapy. The overall response rates were 50% for Regimen A and 65% for Regimen B (P less than 0.05). The complete response rates were 18% for Regimen A and 44% for Regimen B (P less than 0.01). For patients with limited disease, the complete responders were 35% on Regimen A and 52% on Regimen B (P = 0.26); for those with extensive disease, the complete responders were 0% on Regimen A and 35% on Regimen B (P = 0.002). The median survival for complete responders was 17 months on Regimen A and 20 months on Regimen B. The difference is not statistically significant. Toxicity was tolerable for both groups; however, it was greater for the etoposide arm. We conclude that although etoposide improves the overall response rates in patients with small cell lung cancer, especially those with extensive disease, the addition of this drug does not lead to improved survival. PMID- 3024829 TI - Teniposide in the treatment of non-small cell lung carcinoma. AB - Forty-two evaluable patients with advanced non-small cell lung cancer were treated with teniposide at doses ranging from 120 to 180 mg/m2 on Days 1, 3, and 5 every 3 weeks. Thirty-four patients had received no prior chemotherapy. Seven partial responses (16.6%) were obtained (21% in chemotherapy-unexposed patients). Marrow toxicity was the main side effect: life-threatening thrombocytopenia occurred in 9% of patients, and 54.5% experienced severe leukopenia. Teniposide, at the doses and schedule employed, has moderate activity in non-small cell lung cancer. PMID- 3024830 TI - Treatment of non-small cell lung cancer with coumarin and cimetidine. PMID- 3024831 TI - Synthesis of 5-amino-5-deoxy-D-galactopyranose and 1,5-dideoxy-1,5-imino-D galactitol, and their inhibition of alpha- and beta-D-galactosidases. AB - A 12-step route is presented starting from 1,2:5,6-di-O-isopropylidene-alpha-D glucofuranose for the preparation of the title compounds and their L-altro analogues. Their synthesis is based on the reduction with Raney nickel of a protected 5-hydroxyimino derivative of L-arabino-hexofuranos-5-ulose, with the following improvements for the preparation of a D-galactofuranose derivative: oxidation at C-3 with pyridinium dichromate-acetic anhydride, stereospecific reduction of a 3-O-acetyl-hex-3-enofuranose intermediate to the D-gulo derivative, and inversion at C-3 of its 3-tosylate with tetrabutylammonium acetate in chlorobenzene. alpha-D-Galactosidase from coffee beans and from Escherichia coli and beta-D-galactosidase from E. coli and Aspergillus wentii were inhibited with Ki values that ranged from 0.0007 to 8.2 microM. Formation of the enzyme-inhibitor complexes with the D-galactose analogue was on the time scale of minutes, whereas the D-galactitol analogue showed a slow approach to the inhibition only with alpha-D-galactosidase from coffee beans and beta-D galactosidase from A. wentii. N-Alkylation of the D-galactitol analogue was detrimental to the inhibition except for beta-D-galactosidase from E. coli and beta-D-glucosidase from almonds, but, even with these enzymes, the observed affinity enhancements were 10(2) to 10(3)-times smaller than those of N-alkylated D-galactosylamine and D-glucosylamine. PMID- 3024833 TI - The effect of Kupffer cell stimulation or depression on the development of liver metastases in the rat. AB - Tumour cells from a squamous carcinoma (approximately 2.5 X 10(5)) were injected intraportally into a syngeneic strain of rats to produce liver metastases 14 days later. Kupffer cells were stimulated by Corynebacterium parvum (7 mg or 1 mg i.v.) and zymosan (10 mg intraportally). Kupffer cell activity was depressed by the administration of silica, gadolinium chloride or human red cells. The animals in each group were sacrificed at 14 days, the livers removed and the number of visible surface metastases counted and compared. (Mann-Whitney U-test). Kupffer cell stimulation significantly reduced the number of surface liver metastases in all animals (P 0.0048). In contrast depression of Kupffer cell activity significantly increased the number of metastases in all animals (P 0.0045), suggesting that the activity of these cells has an important effect on the development of liver metastases. PMID- 3024832 TI - Clonal analysis and in situ characterization of lymphocytes infiltrating human breast carcinomas. AB - T lymphocytes were isolated from tumor biopsies in 13 patients with breast carcinomas. Immunohistology with monoclonal antibodies confirmed the presence of mononuclear cell infiltrates composed primarily of T lymphocytes in all tumors studied. While the proportion of T lymphocytes expressing the T4 or the T8 surface marker varied from tumor to tumor as determined by morphometric analysis, T8+ cells were more numerous than T4+ cells in 8/12 breast tumors studied. Relatively few T cells (less than 10% in 11/12 tumors) were in an activated state as judged by the surface expression of HLA-DR antigens or the receptor for interleukin-2 (IL-2). In 1 case 20% of the infiltrating mononuclear cells were expressing the IL-2 receptor. The tumor infiltrating lymphocytes (TIL) recovered from 10 tumors were cloned in a microculture system that permits proliferation of nearly 100% of normal peripheral blood T lymphocytes (PBL-T). In contrast to normal and autologous PBL-T, frequencies of proliferating T lymphocyte precursors (PTL-P) were depressed (less than 0.01) in 7/10 TIL preparations indicating a decreased responsiveness of TIL to phytohemagglutinin at the single-cell level. The frequency of PTL-P was noticeably higher in 2 cases (0.03 and 0.09) and close to normal in 1 case (0.39). A total of 170 clones were expanded in vitro and analyzed for different functional capabilities. Most of these clones expressed the T4+/T8-phenotype (73%) and strikingly 53% of these T4+/T8- clones were cytolytic in a lectin-dependent assay, a functional subset which is uncommon among normal PBL-T. Some clones (10%) lysed allogeneic breast tumor cells (MCF7). Only 15% of the clones displayed natural killer activity. Among the cytolytic clones, 17 of 31 tested were also IL-2 producers irrespective of the T4 or T8 phenotype. Our results show that human mammary carcinomas contain many infiltrating T cells with cytolytic potential. Interestingly, among the proliferating cytolytic T cell clones (56% of the microcultures), many expressed the T4+/T8- phenotype. These findings may indicate that the in situ cytolytic reaction (against unknown antigens) is associated preferentially with class II antigens. PMID- 3024834 TI - [Evaluation of the effectiveness of an angiotensin-converting enzyme inhibitor with various methods of continuous recording of blood pressure]. PMID- 3024835 TI - Animal model of silent myocarditis in athymic mice. AB - To clarify the role of the immune system in the development of myocarditis, BALB/c-nu/nu mice (group 1), BALB/c-nu/+ mice (group 2), BALB/c-nu/nu mice injected with 5 X 10(7) spleen cells from BALB/c-nu/+ mice (group 3), and BALB/c nu/nu mice injected with 5 X 10(7) spleen cells from BALB/c-nu/+ mice treated with rat anti-Thy-1.2 monoclonal antibody with complement (group 4) were inoculated with encephalomyocarditis virus. There were no significant differences in the incidence of myocarditis among the four groups. Virus titrations of the heart and serum neutralising antibody titres in the four groups did not show any significant differences. Fifty two per cent (26/50) of group 2 and 43% (20/46) of group 3 died on days 9-15, when congestive heart failure developed. Only 9% (5/54) of group 1 and 8% (1/12) of group 4, however, died on days 9-15. Pathological examination confirmed congestive heart failure in groups 2 and 3 but not in groups 1 and 4. Dilatation of the ventricular cavities, pleural effusion, ascites, and congestion of the lungs and liver were present in groups 2 and 3 but not in groups 1 and 4. Cellular infiltration and myocardial necrosis were severe in groups 2 and 3 but minimal in groups 1 and 4. Thus the severity of myocarditis may be regulated by T cells. So-called silent myocarditis seen in clinical settings may be similar to myocarditis in BALB/c-nu/nu mice. PMID- 3024836 TI - Centrally induced cardiovascular and sympathetic responses to adrenocorticotrophic hormone. AB - To investigate the effects of adrenocorticotrophic hormone on central cardiovascular regulation, an intracerebroventricular injection of the drug was given to male Wistar rats. With doses of 1 microgram and 10 micrograms per rat, blood pressure began to rise within 1 min, attaining a maximum value 5-10 min later. Both heart rate and abdominal sympathetic nerve activity increased simultaneously with the rise in blood pressure. Injection into the fourth ventricle or intravenous administration of the drug elicited no appreciable cardiovascular responses. These results suggest that endogenous adrenocorticotrophic hormone produced locally in the hypothalamus may participate in central cardiovascular regulation by increasing sympathetic outflow. PMID- 3024837 TI - Histochemical examination of energy metabolism in aortic vein grafts in rats. AB - The duration of the presumed metabolic depression of syngeneic vena cava to aorta transplants was determined in rats and the site and type of energy metabolism in the vein grafts assessed. The aerobic metabolic activity was measured from the histochemical reactivity of the enzymes, succinate dehydrogenase and cytochrome oxidase, and the anaerobic activity by staining with lactate dehydrogenase. The activity of the hexose-monophosphate shunt was assessed by the histochemical demonstration of glucose-6-phosphate dehydrogenase. Sixteen hours after grafting a pronounced metabolic depression was noted. Recovery occurred 24 hours after transplantation. The most intense staining was from lactate dehydrogenase in the vein grafts and in the non-transplanted veins. At the end of the observation period of four months the grafts were definitely more strongly stained than the non-transplanted veins, with most of the activity in the thickened intima. This layer had a metabolic profile resembling that of the media of the adjacent aorta. PMID- 3024838 TI - [The effect of a fiber-rich diet on glucoregulation and lipidemia in diabetics]. PMID- 3024839 TI - Adrenergic receptors and the control of cell division in the intestinal epithelium. PMID- 3024840 TI - Fate of annular gap junctions in the papillary cells of the enamel organ in the rat incisor. AB - To investigate the mechanisms whereby annular gap junctions in the papillary cells of the enamel organ are degraded intracellularly, continuously growing rat incisors were examined by electron microscopy of routine thin sections as well as for the cytochemical localization of inorganic trimetaphosphatase activity. Routine thin-section analysis revealed small flat or undulated gap junctions, hemi-annular gap junctions between an invaginated cell process and a cell body, and fully internalized cytoplasmic annular gap junctions. Both hemi-annular and annular gap junctions usually contain various organelles and/or inclusions, such as mitochondria, endoplasmic reticulum, ribosomes, vesicles, and lysosomes in the cytoplasm confined by the junctional membranes. Annular gap junctions are sometimes fused with vesicular or tubulovesicular structures. Cytochemistry of inorganic trimetaphosphatase activity revealed an intense enzymatic reaction within a system of tubular structures and round or oval dense bodies. Both structures are believed to correspond to primary lysosomes. A part of the Golgi apparatus also shows a weak reaction. Although hemi-annular gap junctions never show enzymatic reaction, annular gap junctions sometimes contain reaction products throughout their interior cytoplasm and inclusions. Fusion of annular gap-junctional membranes with reaction-positive tubular structures is also observed. In one instance, revealed in serial sections, an annular gap junction was encircled entirely by a reaction-positive structure. These results suggest that cytoplasmic annular gap junctions are formed by endocytosis of hemi-annular gap junctional membranes from the cell surface and then degraded intracellularly by lysosomal enzymes. PMID- 3024841 TI - Ultracytochemical evidence for the presence of GERL in pinealocytes of the Mongolian gerbil (Meriones unguiculatus). AB - Ultracytochemical reactions for the demonstration of acid phosphatase, glucose-6 phosphatase and thiamine pyrophosphatase, as well as zinc iodide-osmium tetroxide impregnation, revealed the existence of GERL (Golgi apparatus-Endoplasmic Reticulum-Lysosomes) in pinealocytes of the Mongolian gerbil (Meriones unguiculatus). The spatial arrangement of this structure was studied on thick sections using a goniometric stage. Although it was not possible to determine whether GERL in pinealocytes belongs to the Golgi apparatus or to endoplasmic reticulum, it can be concluded that its presence in studied cells signifies that they are considerably more active synthetically than has been believed to date. PMID- 3024842 TI - [A study on the rotavirus antibody in the normal human sera using ELISA]. PMID- 3024843 TI - Arachidonic acid metabolism in inflammation and hypersensitivity reactions: a brief introduction. PMID- 3024844 TI - Regulation of pain sensitivity, influence of prostaglandins. PMID- 3024845 TI - Clinical aspects of prostaglandins and leukotrienes in migraine. PMID- 3024846 TI - Eukaryotic DNA replication: a complex picture partially clarified. PMID- 3024847 TI - A cellular DNA-binding protein that activates eukaryotic transcription and DNA replication. AB - Transcription factor CTF, which is responsible for selective recognition of eukaryotic promoters that contain the sequence CCAAT, was purified to apparent homogeneity by sequence-specific DNA affinity chromatography. Binding sites for CTF in the human Ha-ras and alpha-globin promoters were highly homologous to sequences recognized by nuclear factor I (NF-I), a cellular DNA-binding protein that is required for the initiation of adenovirus DNA replication in vitro. To determine the relationship between CTF and NF-I, we compared the biochemical properties of these two proteins. CTF and NF-I were found to be indistinguishable in polypeptide composition, DNA-binding properties, immunological cross reactivity, and in vitro stimulation of DNA replication and transcription initiation. We conclude that CTF/NF-I can serve both as a transcription selectivity factor for RNA polymerase II and as an initiation factor for adenovirus DNA replication. PMID- 3024849 TI - The type of human papillomavirus present in cervical infections can be determined by the occurrence of specific marker proteins. AB - Four different proteins have been identified on high resolution two dimensional gels of [35S] methionine labelled human cervical biopsies whose expression correlates with the presence of papillomavirus. They are all basic proteins having molecular weights in the region of 48 to 50 kd and are normally expressed individually in different lesions unless the lesion results from a co-infection of two virus types. Comparison of the occurrence of these marker proteins with the actual HPV type present, determined by in situ filter hybridisation, has shown that two are found exclusively with HPV types 6/11 while the other two are found with types 16/18. PMID- 3024848 TI - Comparison of the effect of scorpion venom (Buthus martensii Kashi) on the rat brain and heart mitochondria. AB - A partially purified fraction SVc and a purified homogeneous polypeptide SVIII were isolated from the scorpion (Buthus martensii Kashi) venom, collected in Shan Dong Province of China. SVc decreased the RCR, ADP/O and Qo2 of the rat brain mitochondria. It also decreased the cytochrome oxidase activity and increased the membrane lipid fluidity of the mitochondria. Effect of scorpion venom on the rat heart mitochondria was somewhat different from that of rat brain mitochondria. SVc also decreased RCR, ADP/O and increased the membrane lipid fluidity of heart mitochondria. However, the Qo2 and cytochrome oxidase activity were increased. SVIII has a similar effect on the rat brain and heart mitochondria, but its concentration used is only 1/10 of the effective concentration of SVc. PMID- 3024850 TI - [Classification of germinal tumors of the testis]. AB - Based on the collection of more than 500 personal as well as consiliary cases of TGCT and a thorough review of the existing classifications, the essential synonyms used for designating TGCT entities have been subjected to a critical reappraisal. The tumour entities have been defined exactly to facilitate their coding according to the ICD-O system. The existing classification system have been subjected to a critical reappraisal and compared one to another as well as to our own classification as suggested by ourselves in 1974 and used since then. A detailed discussion and exact coding of well-defined entities is believed to permit an almost complete interconvertibility of the most significant classifications with that of ours and thus to form a reliable basis for amy form of comparison, including the therapeutic results. The bioptic examination of surgical specimens obtained by orchiectomy (other samples are regarded unsuitable) must be carried out in a standardized manner. This should permit not only the usual typing of the respective tumour but also the data necessary for proper tumour staging which today appears to be a natural prerequisite for rational treatment and its monitoring. The main principles of bioptic examination of testicular tumours have been summarized in brief. PMID- 3024851 TI - Cytoxan, epirubicin, methotrexate and 5-fluorouracil with hormonal synchronization (tamoxifen/premarin) in advanced breast cancer. Preliminary results. AB - Thirty-three patients with advanced breast cancer were treated with combination chemotherapy and hormonal synchronization in an attempt to increase the overall response rate generally obtained with traditional modalities of treatment. Among the 31 evaluable patients 1 complete (3%), 7 partial (22%), 13 stable disease (42%) and 10 progression of disease (53%) were obtained. Side effects were quite manageable although there were two episodes of life-threatening hematological toxicity. Taking into account the poor prognostic characteristics of our patients (high percentage of dominant visceral disease, 21/31 previously treated patients), our preliminary results with this regimen are interesting and the treatment deserves further evaluation. PMID- 3024853 TI - Direct electron spin resonance detection of free radical intermediates during the peroxidase catalyzed oxidation of phenacetin metabolites. AB - The oxidation of the phenacetin metabolites p-phenetidine and acetaminophen by peroxidases was investigated. Free radical intermediates from both metabolites were detected using fast-flow ESR spectroscopy. Oxidation of acetaminophen with either lactoperoxidase and hydrogen peroxide or horseradish peroxidase and hydrogen peroxide resulted in the formation of the N-acetyl-4-aminophenoxyl free radical. Totally resolved spectra were obtained and completely analyzed. The radical concentration was dependent on the square root of the enzyme concentration, indicating second-order decay of the radical, as is consistent with its dimerization or disproportionation. The horseradish peroxidase/hydrogen peroxide-catalyzed oxidation of p-phenetidine (4-ethoxyaniline) at pH 7.5-8.5 resulted in the one-electron oxidation products, the 4-ethoxyaniline cation free radical. The ESR spectra were well resolved and could be unambiguously assigned. Again, the enzyme dependence of the radical concentration indicated a second order decay. The ESR spectrum of the conjugate base of the 4-ethoxyaniline cation radical, the neutral 4-ethoxyphenazyl free radical, was obtained at pH 11-12 by the oxidation of p-phenetidine with potassium permanganate. PMID- 3024852 TI - Combination chemotherapy for non small cell lung cancer stage III M0-1 with cisplatin and vinblastine in a divided-dose schedule. AB - Twenty-nine patients with advanced non small cell lung cancer (NSCLC) were treated with a combination of high-dose cisplatin and divided-dose vinblastine. In 27 evaluable patients 15% reached partial response and 59% stable disease. Extension of disease, histological type, performance status and weight loss had no relationship to response. Median duration of response was 10.2 months with a median survival time of 15.4 months in responding patients compared with 14 months of stable disease (p:n.s.) and 4.8 months of progressive disease (p less than 0.001). Gastrointestinal, neurological side effects and the development of a severe debilitation syndrome were the most troublesome toxicities of this treatment. The regimen is not generally suitable for treatment of advanced NSCLC. PMID- 3024854 TI - Generation of superoxide radical and hydrogen peroxide by 1,2,4-triaminobenzene, a mutagenic and myotoxic aromatic amine. AB - 1,2,4-Triaminobenzene, the myotoxic and mutagenic metabolite of several azo dyes, has been shown to generate superoxide radical and hydrogen peroxide during its autoxidation in vitro. Hydrogen peroxide was detected in erythrocytes exposed to the aromatic amine, showing that the autoxidation reaction can occur intracellularly; these cells also suffered oxidative damage, as reflected in glutathione depletion and haemoglobin oxidation. It is suggested that 'active oxygen' species may be involved in the initiation of the toxic changes induced by 1,2,4-triaminobenzene. PMID- 3024855 TI - The Ca2+ antagonist activity of lignans. PMID- 3024856 TI - Biosynthesis of patulin; in vitro conversion of gentisyl alcohol into patulin by microsomal enzyme(s) and retention of one of the carbinol protons in this reaction. PMID- 3024857 TI - [Chemodectomas of the glomus caroticum]. AB - The authors review the nature, biological evolution, diagnosis and treatment of carotid body tumors both on the basis of data reported in the literature and in the light of their own experience with a population of 11 carotid body chemodectomas. All patients (except one) were subjected to total resection of the neoplasm. In one case, surgery was confined to a partial resection. Operative mortality was nil, and complications consisted in one case of Claude-Bernard Horner syndrome, one lesion of the XII cranial nerve and one lesion of the X cranial nerve. Follow-up of the patients (min. 6 months, max. 10 years) has shown no recurrences or metastases. PMID- 3024858 TI - Efficacy and tolerability of vindesine in the treatment of small-cell lung cancer. A phase II study. AB - Eighteen patients suffering from small-cell lung cancer, all with measurable lesions, were treated with vindesine (3 mg/m2 i.v. on days 1 and 8 of a 21-day cycle). Partial responses (PR) were seen in six patients (three of whom had been previously heavily treated). Most patients tolerated the drug extremely well, an important consideration when expected cure rates are very poor. PMID- 3024859 TI - Superoxide dismutase prevents the thrombin-induced increase in lung vascular permeability: role of superoxide in mediating the alterations in lung fluid balance. AB - We investigated the effects of superoxide dismutase (SOD) and SOD linked to Ficoll (mol. wt = 400,000) on the changes in pulmonary transvascular fluid and protein exchange following pulmonary microembolism induced with alpha-thrombin. Studies were made in chronically prepared unanesthetized sheep with lung lymph fistulas. Control thrombin challenged sheep (n = 5) were compared to animals infused with SOD (the SOD-thrombin group, n = 5) or animals infused with SOD linked to high molecular weight Ficoll (the Ficoll-SOD-thrombin group, n = 6). The Ficoll-SOD-thrombin animals were also compared to animals infused with Ficoll alone (the Ficoll-thrombin group, n = 4). In the control-thrombin group, thrombin induced sustained increases in the pulmonary transvascular protein clearance (pulmonary lymph flow X lymph/plasma protein concentration ratio) and pulmonary vascular resistance (PVR). In the SOD-thrombin group, thrombin initially increased both pulmonary transvascular protein clearance and PVR; however, the later increases in protein clearance and PVR were blunted. The pulmonary reflection coefficients for total protein (sigma), a measure of vascular permeability to protein, decreased from a value of 0.70 +/- 0.03 in normal sheep to 0.60 +/- 0.01 following thrombin challenge (p less than 0.05) indicating an increase in lung vascular permeability. The sigma value in the SOD-treated animals was 0.70 +/- 0.02, indicating a protective effect of SOD. The infusion of the Ficoll-SOD complex also attenuated the increases in pulmonary transvascular protein clearance and PVR after thrombin. However, the infusion of Ficoll alone induced a similar protection. The lymph from the SOD-thrombin and Ficoll-SOD thrombin groups prevented the reduction of ferricytochrome C by xanthine/xanthine oxidase, whereas, the lymph from the Ficoll-thrombin animals did not have this effect, indicating SOD activity was present in the animals receiving the enzyme but not in the group infused with Ficoll alone. Differences in the degree of intravascular coagulation could not explain the response to Ficoll since the decreases in fibrinogen concentration following the thrombin were similar in all the groups. Since Ficoll and related dextrans may modify neutrophil function, in particular neutrophil adherence to the endothelium, we examined the effects of Ficoll on neutrophil adherence. The results indicated that when Ficoll was added to the endothelial medium Ficoll reduced the increase adherence of neutrophils to the endothelial cell monolayer. Therefore, Ficoll as a carrier for SOD may provide a direct protection in models of lung vascular injury that are dependent on neutrophils.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3024860 TI - Immediate effect of expiratory loading on left ventricular stroke volume. AB - While the steady-state effects of positive pleural pressure on the circulation have been extensively studied, less is known about the immediate effects of positive intrathoracic pressure on cardiac dynamics. Therefore, we performed electrocardiographically gated radionuclide ventriculography with a respiratory gating technique in nine healthy subjects during quiet breathing and during expiration against a 24 cm H2O expiratory threshold load. During expiration, respiratory loading caused an increase in stroke counts by 29.4% (p less than .001) due to an increase in end-diastolic counts of 26.1% (p less than .001). End systolic counts also rose 18.8% (p less than .05). The ejection fraction did not change significantly. These findings indicate that the increase in left ventricular stroke volume that occurs during the first 1 or 2 beats of a loaded expiration is due to an increase in left ventricular filling and not to augmentation of left ventricular ejection. This immediate increase in pulmonary venous return may reflect increased distensibility of the left ventricle due to decreased filling of the right ventricle. PMID- 3024861 TI - Nuclear magnetic resonance imaging: current applications and future prospects. PMID- 3024862 TI - Adrenergic coronary vasoconstriction during myocardial underperfusion. AB - The effects of adrenergic alpha-receptor-mediated coronary vasoconstriction were examined in the presence of coronary stenosis and during experimental coronary hypoperfusion. Adrenergic coronary vasoconstriction was observed to operate even in the presence of coronary stenosis to limit oxygen delivery to the heart and increase myocardial oxygen extraction, but this vasoconstrictor effect did not result in myocardial lactate production. During constant coronary pressure perfusion the transmural distribution of alpha-receptor coronary vasoconstriction was determined with microspheres. A uniform transmural distribution of alpha receptor-mediated vasoconstriction was observed at normal and subnormal perfusion pressures. During constant flow hypoperfusion, which permits transmural coronary steal, the left ventricular inner/outer blood flow ratio was more favorable with alpha-receptors intact than with alpha-receptors blocked. These findings indicate that alpha-receptor-mediated coronary vasoconstriction has an unexpected beneficial effect by lessening transmural steal during coronary hypoperfusion. PMID- 3024863 TI - Converting-enzyme inhibition and coronary blood flow. AB - The effects of converting-enzyme inhibition (25 mg oral captopril) on coronary hemodynamics in the presence and absence of activation of the renin-angiotension system were studied in 10 patients with mild essential hypertension with no evidence of ischemic heart disease. Coronary blood flow was determined by thermodilution before and after 1 week of diuretic therapy in six patients and before and after placebo in four patients. The diuretic (50 mg/day furosemide) reduced coronary blood flow and increased coronary vascular resistance; in these same patients, captopril reduced mean arterial pressure and the rate-pressure product but increased coronary flow significantly. There was no change in any of these variables after captopril in the placebo group. Similar results were obtained in normotensive rats treated with hydrochlorothiazide; the increase in coronary flow after captopril correlated significantly with the control plasma renin activity. These results in hypertensive humans and normotensive experimental animals indicate that diuretic therapy reduces coronary blood flow significantly, and angiotensin can play a significant role in modulating coronary vascular resistance under conditions associated with activation of the renin angiotensin system. PMID- 3024864 TI - A trial comparing nedocromil sodium (Tilade) and placebo in the management of bronchial asthma. AB - In a double-blind group comparative trial nedocromil sodium (Tilade) at a dose of 4 mg four times daily was compared with placebo in the management of out-patients with bronchial asthma. Treatments were delivered by pressurized aerosol over a period of 28 days following a 2-week base-line during which patients continued on their usual therapy. Twenty-one patients entered the nedocromil sodium group and twenty entered the placebo group. All were using beclomethasone dipropionate aerosol as maintenance steroid therapy plus intermittent use of a bronchodilator taken by inhalation. The dose of steroid was reduced for all patients after 2 weeks of treatment and again for approximately half the patients after 3 weeks trial treatment. Patients in the nedocromil sodium treatment group improved in respect of Diary Card symptom scores and peak expiratory flow rate (PEFR), and in their requirements for inhaled bronchodilators. Patients in the placebo group were worse, particularly in respect of daytime asthma symptoms (P less than 0.01), bronchodilator use (P less than 0.05) and morning PEFR during the third week of trial treatment (P less than 0.05). More patients in the nedocromil sodium group than in the placebo group thought their treatment had been effective (P less than 0.05). Nedocromil was well tolerated. Despite the short duration of treatment imposed at this stage in the clinical evaluation of a new compound, our results were sufficiently encouraging to prompt further evaluation of nedocromil sodium over the longer period required (3-12 months) for the clinical assessment of a new treatment for chronic asthma. PMID- 3024865 TI - Calcium activated neutral proteases (milli- and micro-CANP) and endogenous CANP inhibitor of muscle in Duchenne muscular dystrophy (DMD). AB - Calcium activated neutral protease (milli- and micro-forms) and its endogenous inhibitor have been quantified in muscle from Duchenne muscular dystrophy (DMD) patients. The specific activities of both the enzymes are found to be significantly elevated. Some of the properties studied indicate that the enzymes from DMD and normal are not qualitatively different. The ratios of milli- to micro-enzyme in normal and disease state suggest that these enzymes have independent roles to play. The absence of a significant increase in the level of the endogenous inhibitor is probably indicative of its mode of regulation, in disease condition. PMID- 3024866 TI - Loperamide, an opiate analogue, inhibits plasma ACTH levels in patients with Addison's disease. AB - The effects of loperamide, an opiate analogue of the piperidine class on pituitary hormone secretion were evaluated in eight patients with Addison's disease. In all patients loperamide administration (16 mg orally) induced a marked fall in plasma ACTH levels (P less than 0.01), without affecting GH, PRL and LH levels. Plasma ACTH concentration fell significantly from 854 +/- 167 pg/ml (mean +/- SEM) to 460 +/- 123 pg/ml at 60 min (P less than 0.01). The inhibition persisted throughout the whole test period, the nadir being reached at 300 min. Low dose naloxone infusion 180 min after loperamide administration caused plasma ACTH to rise from 181 +/- 61 pg/ml to 539 +/- 99 pg/ml (P less than 0.01). The present data suggest that the opiate analogue loperamide is a potent inhibitor of ACTH secretion in patients with Addison's disease, which may be acting on mu receptors, since its effect is blocked by low doses of naloxone. PMID- 3024867 TI - ACTH and CRF-producing bronchial carcinoid associated with Cushing's syndrome. AB - A young female patient, with clinical and biochemical manifestations of severe hypercorticism and with the presence of a pituitary adenoma shown by computerized tomography, was thought to have Cushing's syndrome of hypophysial origin. However, the surgically-removed pituitary adenoma contained no ACTH, by immunocytology, and hypercorticism persisted after transsphenoidal adenomectomy. The patient died and autopsy demonstrated an ACTH and corticotrophin releasing factor (CRF)-containing bronchial carcinoid. It can be concluded that bronchial carcinoids can produce ACTH and CRF and can mimic the clinical and biochemical manifestations of pituitary Cushing's syndrome. Thus, the localization of the primary site of hypercorticism can be extremely difficult in patients who have an insidious, occult extrapituitary tumour. Further work is required to establish whether CRF plays a role in the causation of Cushing's syndrome and whether the simultaneous secretion of this peptide can modify the clinical and biochemical manifestations of the ectopic ACTH syndrome. PMID- 3024868 TI - Long-term effect of iodized oil on serum thyroglobulin levels in endemic goitre patients. AB - Serum thyroglobulin (Tg) response to bovine TSH (bTSH) was evaluated in 44 goitrous patients (grades III and IV) living in conditions of chronic iodine (I) deficiency (iodine urinary excretion less than 40 micrograms I/g) and in 26 normal subjects. After the initial clinical evaluation and laboratory tests (bTSH test, T4, T3, anti-Tg and anti-microsomal antibodies) all goitrous patients received 1 ml i.m. of iodized oil (I-oil) and were followed up for 30 months. The bTSH test was repeated at 6, 12, 20 and 30 months after I-oil in 21 subjects. A marked reduction in goitre size was observed in 85% of the patients with a concomitant significant increase in the mean serum T4 and T3 concentrations, a significant fall in the mean serum TSH level and a significant decrease in the T3/T4 ratio. Goitrous patients had elevated serum basal Tg levels (55 +/- 8 SEM micrograms/l) and a significantly mean higher peak Tg value after bTSH (200 +/- 65 micrograms/l) as compared with normal subjects (respectively, 11 +/- 1.4 and 32 +/- 3.4 micrograms/l). Larger goitres (grade IV) had a significantly higher mean peak Tg response as compared with grade III goitres. Treatment with I-oil significantly reduced the mean peak Tg response to bTSH after 6 months (59 +/- 10 micrograms/l) but at 12 and 20 months the peak Tg response after the injection rose, respectively, to 110 +/- 19 micrograms/l and 92 +/- 14 micrograms/l (P less than 0.02 as compared with 6 months), returning to the normal range only at 30 months.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3024869 TI - The effect of cholinergic blockade on the ACTH, beta-endorphin and cortisol responses to insulin-induced hypoglycaemia. AB - To assess the effect of cholinergic blockade on the ACTH, beta-endorphin and cortisol responses to insulin-induced hypoglycaemia, six healthy male volunteers each underwent two insulin tolerance tests in random order, separated by at least 1 week with and without atropine. ACTH levels were significantly greater at +45 min (mean +/- SEM, 223 +/- 21 pg/ml vs 148 +/- 15 pg/ml, P less than 0.01) and at +120 min (54 +/- 11 pg/ml vs 29 +/- 10 pg/ml, P less than 0.05). beta-endorphin levels were significantly greater at +30 min (170 +/- 45 pg/ml vs 96 +/- 32 pg/ml, P less than 0.05) and at +105 min (81 +/- 14 pg/ml vs 54 +/- 7 pg/ml, P less than 0.01). Cholinergic blockade had no effect on plasma glucose or cortisol concentrations. This study demonstrates that cholinergic blockade with atropine facilitates the ACTH and beta-endorphin responses to insulin-induced hypoglycaemia without altering the cortisol responses. PMID- 3024870 TI - Diagnosis and management of ACTH-dependent Cushing's syndrome: comparison of the features in ectopic and pituitary ACTH production. AB - The clinical features, diagnosis and management of 16 consecutive patients with ectopic ACTH production are described and biochemical data are compared with those of 48 consecutive patients with pituitary-dependent Cushing's disease. In 10 cases the ectopic ACTH secreting tumour was completely occult to routine clinical and radiological investigation, and no basal or dynamic investigation of adrenal-pituitary function was able clearly to differentiate these patients from those with Cushing's disease. High dose dexamethasone suppression testing assessed by plasma cortisol was usually helpful but unexpected responses were seen in both diagnostic groups; the metyrapone test yielded no useful information and should now be abandoned. Hypokalaemia was seen in all patients with ectopic ACTH production but in only 10% of those with Cushing's disease who were not on diuretics at presentation. Successful diagnosis and tumour localization was most frequently achieved by a combination of CT scanning of the chest and abdomen and venous catheter sampling for ACTH. All patients in whom the ectopic ACTH secreting tumour was obvious at presentation died of their primary tumour within 8 months, whereas seven of the 10 patients with occult tumours at presentation are alive 1.5-16.5 years later, and appear cured. Occult ectopic ACTH secretion may be impossible to distinguish from pituitary Cushing's disease. Multiple and repeated investigations are often required to make this differential diagnosis, essential for appropriate therapy. PMID- 3024871 TI - A study of corticotroph adenomas in Cushing's disease: no evidence of intermediate lobe origin. AB - There is little evidence for a separate functional or anatomical intermediate lobe in the adult human pituitary gland. Nevertheless, Lamberts et al. (1982) proposed that a subgroup of corticotroph adenomas in Cushing's disease arise in that lobe and can be identified by the presence of argyrophil (? neural) fibres, and that these tumours are more often associated with corticotroph hyperplasia and hyperprolactinaemia than those arising in the anterior lobe. We have examined a series of corticotroph adenomas from patients with Cushing's disease for evidence of argyrophil fibres, and have correlated this with tumour site, corticotroph distribution in the para-adenomatous gland, serum PRL levels and PRL immunoreactive cells in the tumour. Argyrophil fibres were identified not only in tumours adjacent to the posterior lobe, but also in tumours situated deep in the anterior lobe. There was no correlation between the presence of fibres or the site of the tumour and corticotroph hyperplasia. Whilst the two patients with the highest serum PRL levels did have argyrophil fibres they also had a subpopulation of PRL immunoreactive cells in the tumour. On the basis of these results, we propose that the 'intermediate lobe' hypothesis as outlined above should not be accepted. PMID- 3024872 TI - The immunosuppressive effect of methimazole on cell-mediated immunity is mediated by its capacity to inhibit peroxidase and to scavenge free oxygen radicals. AB - We have investigated the effect of methimazole (MMI) on cell-mediated immunity and ascertained the mechanisms of immunosuppression produced by the drug. Methimazole (greater than or equal to 10(-5) M) produced a dose-dependent inhibition in 'active' (early) rosette formation with sheep red cells and in phytohaemagglutinin (PHA)-induced lymphocyte transformation. A concentration of 10(-4) M MMI inhibited the immediate rise in intracellular cAMP triggered by PHA and the subsequent time dependent decrement over 24 h. The drug (10(-3) M) also exerted a significant inhibitory effect on antibody-dependent cell-mediated cytotoxicity (ADCC) over six-fold difference in target/effector cell ratios. At a concentration of 10(-5) M, MMI inhibited zymosan-induced respiratory burst (determined by change in the chemiluminescence of oxidized luminol) in polymorphonuclear and mononuclear cell preparations. Ninety-five per cent of the chemiluminescence in the latter preparation was due to monocytes. At concentrations between 10(-7) and 10(-6) M, MMI significantly inhibited (in cell free systems) horseradish peroxidase-dependent generation of chemiluminescence as well as the oxidation of luminol by hydrogen peroxide. Methimazole exerts its inhibitory effects on measures of cell-mediated immunity by at least two mechanisms: inhibition of peroxidase and scavenging free oxygen radicals. Insensitivity of the test systems or poor access of MMI to leucocytes may account for the need for greater than or equal to 10(-5) M MMI to inhibit cell-mediated immunity significantly. PMID- 3024873 TI - The effect of ovine corticotrophin releasing factor (oCRF), bromocriptine and TRH on the secretion of ACTH and alpha-MSH in Nelson's syndrome and Cushing's disease. AB - The circulating levels of ACTH and alpha-melanocyte stimulating hormone (alpha MSH) were measured in 9 patients with Nelson's syndrome after the administration of saline, ovine corticotrophin releasing factor (oCRF), bromocriptine or TRH. The concentrations of ACTH were grossly elevated and alpha-MSH levels ranged from undetectable to higher than the normal range. In seven of eight subjects there was a rapid corticotrophic response, but no change in the alpha-MSH level, following oCRF. This response was delayed in one subject. Following oCRF injection, the plasma oCRF profile was variable but circulating oCRF was detectable even at the end of the experiment in all cases. There was no significant change in circulating ACTH or alpha-MSH following either bromocriptine or TRH. Cultured tumour cells from one case of Cushing's disease showed a corticotrophic response but no change in alpha-MSH to oCRF and the response was enhanced by vasopressin. Bromocriptine added to the same tumour depressed ACTH secretion without affecting the output of alpha-MSH. The present data suggest that the tumours in these subjects are responsive to oCRF and arise from corticotrophs rather than melanotrophs. PMID- 3024874 TI - Adrenergic activity and aldosterone regulation: no evidence for an alpha-1 adrenoceptor-mediated influence in normal subjects. AB - In normal man the sympathetic nervous system could exert an inhibitory influence on aldosterone responsiveness to angiotensin II. The possible role of alpha-1 adrenoceptors in the modulation of aldosterone response was assessed by studying the changes of plasma aldosterone during infusion of angiotensin II at the doses of 1, 2, 5 and 10 ng/kg.min or after corticotrophin injection, 0.25 mg, in 9 normal subjects before and after treatment with the selective alpha-1 adrenoceptor antagonist, prazosin. Prazosin, given during 3 weeks, did not modify supine arterial pressure, heart rate and the plasma levels of angiotensin II, renin, aldosterone or adrenaline but caused a significant (P less than 0.05) increase of plasma noradrenaline. The correlation relating plasma aldosterone to plasma angiotensin II levels before and during angiotensin II infusion and the response of plasma aldosterone to corticotrophin was not modified by prazosin. These findings suggest that in normal man there is no inhibitory influence of the noradrenergic system on aldosterone responsiveness to angiotensin II mediated by an alpha-1 dependent mechanism. PMID- 3024875 TI - Studies of hypothalamic pituitary structure and function in patients previously treated with bilateral adrenalectomy alone for Cushing's disease. AB - A controversy still exists in regard to hypothalamic pituitary function long-term after cure of hypercortisolism due to Cushing's disease. In an attempt to resolve this controversy, we have studied 15 patients, treated at least 6 years previously, by bilateral adrenalectomy. None of these patients had had pituitary directed therapy. The maximum increment response of serum TSH in response to TRH was greater than 5 mU/l in 13 of the 15. Serum PRL response to TRH, GH response to insulin-induced hypoglycaemia, gonadotrophin responses to LHRH and nocturnal PRL secretion were normal in all patients studied. When nocturnal GH secretion was corrected for age, body mass index and menopausal status it was definitely abnormal in only two patients. The mean nocturnal GH secretion did not differ from that measured in a control group of Addisonian patients. The series of patients also did not differ significantly from the Addisonian patients in relation to the pattern of changes in plasma ACTH, over 24 h after an 0800 h oral dose of hydrocortisone. There was a significant rise in plasma ACTH between 2200 h and 0600 h in both groups of patients. The plasma ACTH concentrations were significantly higher in post-adrenalectomy patients. Hypothalamic pituitary function is normal in the long-term in the majority of patients treated by bilateral adrenalectomy for Cushing's disease. PMID- 3024876 TI - Pituitary pro-opiomelanocortin-cell carcinoma occurring in conjunction with a glioblastoma in a patient with Cushing's disease and subsequent Nelson's syndrome. AB - Pituitary carcinoma is defined as a malignant pituitary tumour associated with blood- or lymph-borne metastases. Cushing's disease is frequently present in patients with this condition. After adrenalectomy for Cushing's disease, a 37 year-old man developed Nelson's syndrome resulting from a pituitary carcinoma with metastases to the spinal cord, cauda equina, heart, liver, and pancreas. The primary tumour and its metastases showed immunocytochemical staining for ACTH, beta-lipotrophin, and variably for beta-endorphin and alpha-melanocyte stimulating hormone (alpha-MSH). A coincidental glioblastoma was also present. Nine cases of Cushing's disease associated with pituitary carcinoma, including the present patient, are documented in the literature. The case reported is only the second in which immunohistochemical staining of the primary pituitary tumour and its metastases was performed, and the first in which ACTH-related peptides, in addition to ACTH itself, were demonstrated in the carcinoma cells. PMID- 3024877 TI - Alpha-human atrial natriuretic polypeptide reduces the plasma arginine vasopressin concentration in human subjects. AB - The synthetic alpha-human atrial natriuretic polypeptide (alpha-hANP) was infused into six normotensive, euvolaemic, healthy volunteers to examine the effect on the plasma arginine vasopressin (AVP) concentration. The intravenous administration of alpha-hANP (0.1 microgram/kg/min, 20 min) led to a remarkable reduction in mean blood pressure (-10 mmHg, P less than 0.05), and there was an increase in pulse rate (+10 bpm, P less than 0.05), in each subject. The urinary volume, sodium excretion and cyclic 3',5'-guanosine monophosphate (cyclic-GMP) excretion were increased to 3.5 (P less than 0.05), 2.5 (P less than 0.05) eight fold (P less than 0.01), respectively, during the alpha-hANP infusion. The dose and duration of the synthetic alpha-hANP in the present study was sufficient to induce these cardiovascular and renal effects. The plasma AVP concentrations decreased from 0.39 +/- 0.09 pg/ml to the undetectable level during the alpha hANP administration. After infusion, the plasma concentrations of the AVP promptly returned to preinfusion levels (0.46 +/- 0.14 pg/ml). However, there was no significant change in plasma AVP concentration during placebo infusion. The marked suppression in plasma AVP concentration may account for the remarkable diuresis, in addition of the direct renal effects of the synthetic alpha-hANP. PMID- 3024878 TI - The effect of peripheral catecholamine concentrations on the pituitary-adrenal response to corticotrophin releasing factor in man. AB - To evaluate the effect of changes in plasma catecholamines on the pituitary adrenal response to ovine corticotrophin releasing factor (CRF) in normal man, the response to CRF alone (10 subjects) was compared responses after infusions of adrenaline (6 subjects), noradrenaline (6 subjects) and after oral administration of the alpha 2 agonist clonidine (10 subjects). Compared to control levels, plasma adrenaline and noradrenaline concentrations were increased three- and four fold respectively by exogenous infusions, whereas plasma noradrenaline was significantly lowered by clonidine. Despite these changes in plasma catecholamine levels, the responses of plasma ACTH, cortisol and aldosterone to CRF did not differ significantly from control (CRF alone) in any of the three studies. Neither clonidine pretreatment nor catecholamine infusions altered basal levels of plasma ACTH, cortisol or aldosterone and no effect of CRF or catecholamine manipulations on plasma arginine vasopressin concentration was observed. These results show that modulation of peripheral plasma catecholamine levels within physiological limits does not affect CRF-stimulated release of ACTH or the adrenal response in normal man. PMID- 3024879 TI - Atropine selectively blocks GHRH-induced GH secretion without altering LH, FSH, TSH, PRL and ACTH/cortisol secretion elicited by their specific hypothalamic releasing factors. AB - The role of acetylcholine in the regulation of the hypothalamo-pituitary system in man was assessed using atropine, which selectively blocks cholinergic muscarinic receptors. Paired tests were performed in 10 normal men using either GHRH (1 microgram/kg i.v.), or TRH (300 micrograms i.v.) plus LHRH (100 micrograms i.v.) plus corticotrophin releasing hormone (CRH) (1 microgram/kg i.v.) with or without atropine given 30 min previously (1 mg i.m.). The GHRH induced GH secretory peak (17.8 +/- 3.0 ng/ml) was completely blocked by atropine administration (2.8 +/- 0.6 ng/ml) (P less than 0.05). Atropine did not, however, modify TRH-induced TSH and PRL secretion, nor FSH and LH release induced by the LHRH pulse. ACTH/cortisol secretion elicited by CRH was also unaffected by atropine. These results suggest that atropine blockade of GHRH-induced GH secretion is highly specific, and constitutes an indication of the importance of cholinergic control of GH function. Furthermore, atropine's lack of action on the other pituitary hormones rules out the possibility that it acts non-specifically, i.e. via blood flow changes or toxic effects. PMID- 3024880 TI - Immunological abnormalities in intravenous drug abusers and relationship to the prolonged generalized lymphadenopathy syndrome in Italy. AB - The prolonged generalized lymphadenopathy syndrome (PGL) has been considered a prodromal condition to the Acquired immunodeficiency syndrome (AIDS), but the clinical, virological and immunological characteristics of patients who will develop AIDS are not known. We report on the immunological profile of intravenous drug abusers with or without PGL in Northeastern Italy. We found a reduction of lymphocyte-absolute numbers with reversal of the T4/T8 ratio and decreased Leu 11b+ cells. The response to mitogens and natural killer activity are compromised in PGL patients. Neutrophil function is reduced both in drug abusers with or without lymphadenopathy. The serological investigations revealed a high prevalence of antibodies against HTLV III and the Epstein-Barr viruses. The recognition of immune dysfunction in the intravenous drug abusers appears to be important since these patients develop AIDS and these abnormalities may precede AIDS. PMID- 3024881 TI - Immunological studies on adult T cell leukaemia virus (ATLV) carriers. AB - Adult T cell leukaemia associated antibody (ATLA-Ab) positive people who were considered to be adult T cell leukaemia virus (ATLV) carriers were found in 0.75% of the adult population in the non-endemic area of Nagoya, Japan. Immunological studies on these people revealed that T lymphocyte subpopulations, as defined by nine monoclonal antibodies reactive to T lymphocyte surface antigens including T cell activation antigen, showed no differences between ATLA-Ab positive and ATLA Ab negative individuals. Only a slightly higher percentage of Ia positive T lymphocytes was found in ATLA-Ab positive persons. Furthermore, the serum IgG level and the antibody titre of cytomegalovirus were significantly increased in ATLA-Ab positive individuals, while serum IgA, IgM level and the antibody titre of herpes simplex virus and mumps virus, showed no differences in both groups. This data suggests the possibility that ATLV carriers have some mild immunological abnormalities. PMID- 3024882 TI - Immunomodulating effect of low density lipoprotein on human monocytes. AB - Low density lipoprotein (LDL) isolated from sera of healthy volunteers in 50 micrograms protein/ml concentration induced an early adenylate cyclase activation in human monocytes followed by elevation of cGMP level. In addition, a rapid 45Ca2+ influx was also detected on addition of 25-100 micrograms protein/ml concentrations. The monocyte activating effect of LDL under in vitro circumstances was characterized by an enhanced O2 consumption, H2O2 generation and by the increased release of lysosomal enzymes such as beta-glucuronidase and elastase like protease (ELP). On the other hand, LDL diminished markedly the Fc gamma receptor (Fc gamma R) mediated rosette formation, phagocytosis and the antibody dependent cellular cytotoxicity (ADCC) of monocytes without a significant decrease in the IgG binding capability of cells. High levels of serum LDL may play a significant role in the arterial wall injury by elastase like protease as well as biologically active oxygen species released from monocytes of patients suffering from arteriosclerosis. PMID- 3024883 TI - An antigenic study of human plasma cells in normal tissue and in myeloma: identification of a novel plasma cell associated antigen. AB - A mouse monoclonal antibody named BU11 which detects an antigen strongly expressed on human plasma cells is described. The antibody stains plasma cells in tonsil sections, fresh and cultured plasmacytoid cells from the bone marrow of patients with multiple myeloma and cells of the plasmacytoid cell line RPMI 8226 used as the immunogen. In vitro studies of pokeweed mitogen (PWM) stimulated peripheral blood B cells and Epstein-Barr virus (EBV) stimulated tonsil B cells show that the antigen is present mainly on cells coexpressing the OKT10 antigen and containing cytoplasmic immunoglobulin (cIg). The BU11 antigen is expressed weakly on some normal B cells and is not present on T cells, monocytes or granulocytes. The antigen is of molecular weight 58kD under reducing conditions and is biochemically distinct from previously described plasma cell antigens. PMID- 3024884 TI - In vivo activated cytotoxic T cells in the thyroid infiltrate of patients with Hashimoto's thyroiditis. AB - High proportions of T8+ cells with inverted T4/T8 ratio were found in freshly isolated thyroid lymphocytes from patients with Hashimoto's thyroiditis. In addition, about one third of thyroid infiltrating cells expressed the TAC antigen, whereas in patient peripheral blood (PB) or normal lymphocytes from PB or lymphoid organs the percentage of TAC-positive cells was consistently lower than 10%. Following negative selection with OKT4 or OKT8 monoclonal antibodies and complement, TAC+ T cells were enriched in the T8+ cell population. Thyroid infiltrating T cells from two patients underwent two different cloning procedures. In the first, single T cells were initially activated with phytohaemagglutinin (PHA) and interleukin 2 (IL-2), in the other with recombinant IL-2 (rIL-2) alone. The majority of T cell clones obtained by initial PHA stimulation (55-65%) had the T8+ phenotype, but the frequency of T8+ clones obtained by stimulating T cells with rIL-2 alone was even higher (78 & 71%, respectively). The majority of T8+ clones elicited by PHA (35/37 & 36/38) and all the T8+ clones (36/36 & 22/22) obtained from thyroid infiltrates with initial stimulation by rIL-2 displayed cytolytic activity. Most of cytolytic T8+ clones obtained from thyroid infiltrates with both cloning procedures, displayed NK activity against human K562 and MOLT-4 target cells, but not against a NK resistant target, such as Raji cells. These data suggest that in Hashimoto's disease a considerable proportion of thyroid infiltrating T cells are in vivo activated T8+ cytolytic T cells with NK activity, which may be of importance in determining or maintaining the tissue damage of the target gland. PMID- 3024885 TI - Expression of Tac antigen in B cell lymphomas. AB - In a series of 55 cases of B cell derived non-Hodgkin's lymphoma the reactivity of two distinct anti-Tac monoclonal antibodies was examined using a sensitive immunoperoxidase technique on cryostat sections. Eighteen out of the thirty-five cases of B cell lymphomas of low or intermediate grade of malignancy were found to be reactive while six out of 20 cases of high-grade malignancy lymphomas showed a positive immunostaining. No correlation was found between anti-Tac reactivity and surface immunoglobulin phenotype, T65 antigen, or calla expression. These findings showed that IL2 receptor expression is not restricted to activated T cells, and raise the question of the possible role of IL2 in the regulation of malignant B cell clone expansion. PMID- 3024886 TI - Prognostic value of T lymphocyte subset ratios for renal transplant survival in patients on different immunosuppressive regimens. AB - Previously we reported that the pre-transplant and pre-rejection OKT4/OKT8 ratio can be used to predict renal allograft survival. Patients on azathioprine (Aza) and low-dose steroids (St) with a pretransplant ratio less than or equal to 1.6 exhibited a 6-month graft survival of 33% compared with 79% for those with a ratio greater than 1.6 (P = 0.02). Furthermore, 100% of the rejection episodes treated with high doses of prednisone in patients with a prerejection ratio less than or equal to 1.6 were irreversible in comparison with only 10% for patients with a ratio greater than 1.6 (P less than 0.001). In the present study, we investigated the prognostic value of the OKT4/OKT8 ratio for patients who received rabbit antithymocyte globulin (RATG) as anti-rejection therapy or cyclosporin A (CsA) as basic immunosuppressive therapy. No correlation was found between the pre-transplant OKT4/OKT8 ratio and 6-month graft survival for either treatment group because of an improved graft survival among patients with a pretransplant ratio less than or equal to 1.6 (78% for patients who received RATG and 85% for CsA-treated patients). For Aza-treated patients with an OKT4/OKT8 ratio less than or equal to 1.6 at the time of rejection, rejection episodes that were treated with RATG were reversible in 78% of the cases, whereas among CsA treated patients rejection episodes treated with high doses of prednisone were reversible in 72% of the cases. No significant differences in graft survival or reversibility of rejection episodes between patients with a pre-transplant or prerejection OKT4/OKT8 ratio greater than 1.6 were found. Furthermore, in both the CsA and the Aza-treated patients (with or without RATG), the OKT4/OKT8 ratio had decreased significantly 3 months after transplantation. This decrease was associated with cytomegalovirus infections rather than the type of immunosuppressive therapy. PMID- 3024888 TI - Baroreflex sensitivity alteration following transient hemispheric ischaemia in rats: protective effect of alphamethyldopa and guanfacine. AB - The reflex tachycardia to arterial vasodilation was analysed in anaesthetized and ventilated Long Evans rats. Nicardipine was given by slow intravenous injection. The decrease in blood pressure was accompanied by a sympathetically mediated reflex tachycardia. The slope of the mean blood pressure-pulse period relationship was considered as an index of baroreflex sensitivity. Two injections of nicardipine were given during a single experiment. For studying the suprapontine control of the baroreflex arc, rats were subjected to transient (10 min) bilateral hemispheric ischaemia. These rats exhibited a blood pressure drop following recirculation and baroreflex sensitivity was impaired. Pretreatment with alpha-methyldopa, guanfacine and enalapril lowered mean blood pressure to a similar extent. In those rats pretreated with alpha-methyldopa and guanfacine transient hemispheric ischaemia did not alter baroreflex sensitivity while enalapril pretreated rats exhibited an impaired baroreflex. The protective effect of alpha-methyldopa and guanfacine against the cardiovascular consequence of hemispheric ischaemia may depend on reduced monoamine turnover resulting from central alpha-adrenoceptor stimulation. PMID- 3024887 TI - Release of leukotriene B4 and 5-hydroxyeicosatetraenoic acid during phagocytosis of artificial immune complexes by peripheral neutrophils in chronic inflammatory bowel disease. AB - The capacity of peripheral neutrophils for activation of the arachidonic acid (AA) metabolism was studied during phagocytosis of IgG containing immune complexes (ICs) binding to Fc-receptors. Release of approximately 9% of the intracellular pool of radiolabelled AA in phospholipids, and release of the pro inflammatory mediators, leukotriene B4 (LTB4), constituting 1.8%, and 5 hydroxyeicosatetraenoic acid (5-HETE), constituting 2.9% of the total radioactivity released, were demonstrated in 15 patients with untreated Crohn's disease, 15 patients with ulcerative colitis, and in 15 healthy volunteers. The concentrations of LTB4 and 5-HETE released were within the range of chemotactic activity for the two lipoxygenase products. Multiple large IgG containing ICs were revealed in neutrophils after phagocytosis by immunofluorescence. A minor defect in the IC uptake in patients with Crohn's disease observed in the absence of complement only, did not result in a subnormal activation of arachidonic acid release or metabolism. The study suggests that complexes of the IgG-class previously demonstrated in chronic inflammatory bowel disease, particularly in Crohn's disease, may activate inflammatory neutrophils leading to release of significant amounts of the pro-inflammatory lipoxygenase metabolites, LTB4 and 5 HETE. PMID- 3024889 TI - Pressor sensitivity to angiotensin I and angiotensin II during the development of experimental renal hypertension in the rat. AB - Treatment with the potent angiotensin converting enzyme inhibitor perindopril completely prevented any rise in blood pressure in the 2-kidney, 1-clip (2K1C) model of renal hypertension in rats. Withdrawal of this inhibitor was followed by a slow rise in blood pressure. In 2K1C rats treated with perindopril, pressor responses to angiotensin I fell during the treatment period, but returned to normal after the inhibitor was stopped. Pressor responses to angiotensin II (AII) increased during treatment with perindopril; this was presumably due to increased receptor sensitivity consequent on the falls in endogenous AII levels. Responses to AII fell to control levels after the inhibitor was stopped. It is concluded that an increased pressor sensitivity to AII is not the cause of the slowly developing hypertension in the 2K1C model of hypertension, and that the slow pressor response to AII must be due to other factors. PMID- 3024890 TI - Effect of L-thyroxine on serum angiotensin converting enzyme activity in sheep. AB - The effect of L-thyroxine on serum angiotensin converting enzyme activity using a sheep model was examined. Following two weeks of L-thyroxine treatment, the activity of serum angiotensin converting enzyme was increased from 6.92 U/ml to 8.65 U/ml (s.e.m. = 0.40, n = 13, P less than 0.01). Discontinuation of L thyroxine treatment resulted in lowering the activity of serum angiotensin converting enzyme within three weeks to values close to those observed during the control period. It was concluded that L-thyroxine modulates serum angiotensin converting enzyme activity. The sheep is an appropriate animal model for the study of the factors that control serum angiotensin converting enzyme activity. PMID- 3024891 TI - Decreased 5'-nucleotidase activity in suppressor (OKT8) T lymphocytes from homosexuals with AIDS-related complex: nonassociation with enhanced deoxynucleoside toxicity. AB - The purine metabolic enzymes adenosine deaminase (ADA), purine nucleoside phosphorylase (PNP), and 5'nucleotidase (5NT) have been shown to be important for normal lymphocyte maturation. Abnormalities of these enzymes have been associated with hereditary as well as acquired immunodeficiency states. Enzyme activity was measured in helper (OKT4) and suppressor (OKT8) lymphocyte subsets from 10 homosexuals with AIDS-related complex (ARC) and in 10 healthy controls. There were no significant differences in either mean ADA activity or mean PNP activity between ARC OKT4 cells and control OKT4 cells and between ARC OKT8 cells and control OKT8 cells. By contrast, mean 5NT activity was slightly decreased in OKT4 cells from ARC patients compared with that of controls and more significantly diminished in ARC OKT8 cells compared with that of controls. Both deoxyadenosine and deoxyguanosine, when incubated separately with OKT4 and OKT8 cells in the presence of EHNA, an ADA inhibitor, did not significantly inhibit lymphocyte blastogenesis to a greater extent in ARC patients than in controls. Hence, the decreases in 5NT activity most likely reflect lymphocyte immaturity and are not associated with biochemical abnormalities leading to increased deoxynucleoside toxicity. PMID- 3024893 TI - Effect of LTB4 and its isomers on human leucocyte migration into skin chambers. AB - The in vivo chemotactic effect of LTB4 and of its isomers, 6-trans-LTB4, 12 epi-6 trans-LTB4 and 5S, 12S-DHETE, was tested with a skin chamber technique in healthy volunteers and in parallel in vitro with an under-agarose technique. LTB4 had an in vivo chemotactic effect at 10(-7) mol/l in 24-hour experiments, while its isomers had no in vivo chemotactic effect at this concentration. LTB4 was also in vitro a more effective attractant than its isomers. In addition, C5ades Arg was tested using zymosan-activated serum, and was found to have an in vivo chemotactic effect at 1.5 X 10(-10) mol/l. However, when LTB4 and C5ades Arg were studied in 6-hour experiments in skin chambers there was an alteration in relative potency, LTB4 being relatively more potent at shorter test durations. This is most likely due to metabolisation of LTB4 in the presence of PMN:s and precludes a strict comparison of the in vivo chemotactic effects of LTB4 and C5ades Arg. When zymosan-activated serum or LTB4 was replaced by PBS after six hours in skin chamber experiments more leukocytes accumulated in the chambers at 24 hours than in chambers containing PBS for the whole 24 hour period. The reason for the increased migration even after the removal of the chemo-attractants as well as the relevance of LTB4 and C5a as chemo-attractants in the inflammatory process is discussed. PMID- 3024892 TI - Engagement of adenosine receptors inhibits hydrogen peroxide (H2O2-) release by activated human neutrophils. AB - Adenosine and its analogs, acting at specific cell surface receptors, inhibit generation of superoxide anion by neutrophils. Since it has been suggested that hydrogen peroxide (H2O2) release may not be contingent upon superoxide anion release, we studied the effects of 2-chloroadenosine, a potent adenosine receptor agonist, on the formation of H2O2 by neutrophils exposed to various stimuli: n formyl-methionyl-leucyl-phenylalanine (FMLP), concanavalin A, phorbol myristate acetate (PMA), serum-treated zymosan particles (STZ), and immune complexes. 2 Chloroadenosine (0.01-10 microM) inhibited formation of H2O2 by neutrophils exposed to FMLP, concanavalin A, and STZ particles. As we have found with O2- generation, 2-chloroadenosine failed to inhibit H2O2 release by neutrophils stimulated by either phorbol myristate acetate or immune complexes. The data show that whereas adenosine and its analogs inhibit neutrophil release of H2O2 and superoxide anion in response to most ligands, they fail to inhibit activation of neutrophils by immune complexes. Nor do they inhibit neutrophil activation by PMA, an agent which bypasses cell surface receptors by direct activation of protein kinase C. Surprisingly, we found that adenosine deaminase activity was adsorbed onto zymosan particles during opsonization and enhanced release of H2O2 by neutrophils exposed to STZ. These studies with yeast cell walls suggest that if microorganisms adsorb adenosine deaminase from serum, then the intracellular microbicidal activity of neutrophils is enhanced. PMID- 3024895 TI - [A case of acute polyradiculoneuritis with adenovirus-induced conjunctivitis]. PMID- 3024894 TI - Intracerebral malignant fibrous histiocytoma: a light and electron microscopic study with immunohistochemistry. AB - A 65-year-old man presented with a tumor within the right cerebral hemisphere. Biopsy showed a pleomorphic neoplasm which was diagnosed as a malignant fibrous histiocytoma only after immunohistochemical and electron microscopic examination. Full clinical and radiological investigation, including total body CAT scanning, failed to reveal any other deposits, and thus it is believed to have arisen in the brain. It is emphasised that this diagnosis could not have been reached had only conventional histological techniques been available. PMID- 3024896 TI - Gastric interposition in esophageal carcinoma as a cause of pseudomass on thyroid imaging. PMID- 3024897 TI - Controversies in the management of Cushing's disease. PMID- 3024898 TI - The significance of the pathologic findings in endometrial cancer. AB - Endometrial carcinoma includes several specific subtypes which have differing prognostic implications. The two most common subtypes are adenocarcinoma, not otherwise specified (NOS), and adenoacanthoma. Also included in the adenocarcinoma NOS are secretory carcinoma and mucinous carcinoma which have the same natural history as do adenocarcinomas without these features. Fortunately, the above types have the best prognosis and constitute approximately 80% of all endometrial carcinomas. They also present the major diagnostic problem for the pathologist in distinguishing atypical hyperplasia and some of the metaplasias from well-differentiated carcinoma. The tendency would appear to be the over interpretation of these hyperplastic lesions. Other subtypes have a much less favorable outlook. They include papillary carcinoma, adenosquamous carcinoma, glassy cell carcinoma and clear cell carcinoma. There are two distinct types of papillary carcinoma, the papillary clear cell and the papillary nonclear cell carcinoma. These can readily be separated, and should be, on the basis of prognostic implications. Other pathologic parameters play a significant role in patient management and in the estimation of prognosis. These include postsurgical pathological staging, measurement of depth of myometrial invasion, lymphatic and blood-vascular invasion, serosal involvement, local spread and, perhaps most importantly, tumor grade. Nuclear grading proved to be a better predictor of treatment outcome than did either the FIGO or WHO grading systems. This was especially true in adenocarcinoma NOS, adenoacanthoma and papillary carcinoma. PMID- 3024899 TI - A study of intracellular orthophosphate concentration in human muscle and erythrocytes by 31P nuclear magnetic resonance spectroscopy and selective chemical assay. AB - In order to study the relationship between extracellular and intracellular concentrations of orthophosphate (Pi), phosphorus nuclear magnetic resonance spectra were recorded, at rest, from the flexor digitorum superficialis muscle of hypophosphataemic patients with vitamin D-resistant rickets, and patients with Paget's disease of bone before and after they had been made hyperphosphataemic by treatment with the drug ethylidene-1-hydroxy-1,1-bisphosphonate. Changes in intramuscular P1 were estimated from the ratio of the areas of the Pi to adenosine 5'-triphosphate peaks. Even though the plasma Pi concentration in these patients spanned a fourfold range (0.5-2.0 mmol/l) the corresponding intramuscular Pi concentration increased by only 70%. A similar effect was observed in erythrocytes, from patients with these disorders, which were incubated in autologous plasma at 37 degrees C, under an atmosphere of O2 + CO2 (95:5, v/v). However, chloride ions, which are transported passively across the cell membrane, showed no change in distribution between cells and plasma, indicating that there was no general effect on passive anion distribution. When erythrocytes from normal subjects were incubated in autologous plasma (1.0 mmol of Pi/l) and in plasma supplemented with Pi (2.3 mmol of Pi/l), the Pi concentration in the cells, at steady state, increased only from 0.57 to 0.78 mmol/l cells, suggesting that the effect was not an artifact of disease or drug therapy. It is concluded that, in human skeletal myocytes and erythrocytes, the percentage change in the concentration of cytoplasmic Pi is lower than that in plasma. This implies that these cells can buffer or regulate cytoplasmic Pi when the extracellular concentration is disturbed. PMID- 3024901 TI - The biochemistry of haemostasis. PMID- 3024900 TI - Non-esterified fatty acids may regulate human leucocyte sodium pump activity. AB - Human leucocyte sodium pump activity was studied in normal fasting subjects by measuring the ouabain-sensitive 22Na+ efflux rate constants. This 22Na+ efflux rate constant was inversely related to the fasting plasma non-esterified fatty acid level (rs = -0.73, P less than 0.0001). An oral glucose load (40 g/m2 surface area) led to an increase in the leucocyte ouabain-sensitive 22Na+ efflux rate constant after 2 h (1.97 +/- 0.25 to 2.44 +/- 0.19 h-1, P less than 0.0001, n = 11). There was a concomitant fall in the plasma non-esterified fatty acid level. Incubation of leucocytes in vitro with 100 mumol/l linoleic acid inhibited the leucocyte ouabain-sensitive 22Na+ efflux rate constant (1.52 +/- 0.27 vs 0.84 +/- 0.24 h-1, P less than 0.001, n = 8). The leucocyte Na+,K+-dependent adenosine triphosphatase (Na+,K+-ATPase) activity was inhibited in vitro by long chain non esterified fatty acids, especially when unsaturated. Non-esterified fatty acids may account for some of the Na+,K+-ATPase inhibitory activity of plasma. PMID- 3024902 TI - [Effects of the integration of a habitual diet with glucomannan fiber in hypercholesterolemia. A clinical study in familial and sporadic hyperlipoproteinemia with lipo-proteic phenotype IIa and IIb]. PMID- 3024903 TI - The ten commandments for immunotherapeutic drugs at the example of sulfur containing agents. AB - Immunotherapeutic agents (IMT) are intended to restore or reequilibrate a faltering immune system. Levamisole was the first chemically defined IMT endowed with immunomodulatory effects. In order to clarify the mechanisms of its ambivalent activities, thiocompounds were tested for their ability to modify immune responses. All thiomidazoles are immunomodulatory drugs which dose dependent effectiveness is related with the cholinergic-like influence of the imidazole moiety. The findings suggest also a relationship among thiodrugs between chemical structure and influence on the immune system. These studies lead to the development of sodium diethyldithiocarbamate (Imuthiol), as a non-toxic immunostimulant agent, specifically active on the T-cell lineage. Our studies unequivocally put forth on the specific requirements that are essential to characterize a potential IMT prior to clinical release: in vivo activities no immunosuppression through prolonged administration no antigenicity or hapten-like activity no carcinogenicity or tumor-promoting influence experimental route of administration related to that in humans assays in various animal strains pharmacokinetics chemical characterization known toxicities, teratogenesis, and mutagenesis assays on models of cancer, autoimmune diseases, immunodeficiencies, and infections. The results will allow to determine a range of doses, and predict most direct and secondary effects. They will also specify whether the agent is an adjuvant, a reticulostimulant, an immunomodulatory, an immunostimulatory, an immunorestorative, or an immunoregulating drug. Clinicians will therefore be provided with the data which are essential to perform useful clinical testing. PMID- 3024904 TI - Comparative biochemistry of the ubiquinol-cytochrome c oxidoreductase (EC 1.10.2.2) isolated from different heart mitochondria. AB - The ubiquinol-cytochrome c oxidoreductase (bc1 complex, EC 1.10.2.2) has been isolated from the heart mitochondria of beef, chicken, turkey, duck and tuna with an identical procedure. The polypeptide composition of the different complexes, compared using SDS-polyacrylamide gel electrophoresis, shows that the three subunits carrying the prosthetic groups of the enzyme are highly conserved in all species. Also the large subunits I and II (core proteins) and band VI appear to be conserved in structure, while subunits VII and VIIa show a most remarkable structural variation in the various complexes. The steady-state ubiquinol cytochrome c reductase analysis of the active enzymes indicates that all the bc1 complexes follow essentially a ping-pong mechanism, with the cytochrome c substrate displaying a partial competitive inhibition vs the ubiquinol substrate. The cytochrome c specificity of the reductase activity clearly is different in the various bc1 complexes, whereas the quinol specificity appears to be identical in all the enzymes. PMID- 3024905 TI - Paraganglioma metastatic to the cervical spine. AB - Paragangliomas are usually benign tumors which may be locally invasive. Only 10% of these tumors are malignant. When metastases occur the most common sites are lung, liver and bone. When bony metastases occur the spine is a common site. In the present case cervical spine metastases with resultant bony encroachment and epidural extension of tumor with cord compression was diagnosed with myelography and CT. PMID- 3024906 TI - Antiarrhythmic efficiency of Craviten and changes in the activities of serum dopamine beta-hydroxylase and erythrocyte membrane ATPases. AB - Among 30 patients with ventricular arrhythmia resistant to conventional antiarrhythmic therapy, 33% showed normalization of heart rhythm after single i.v. injection of Craviten at a dose of 6 mg. In all patients, sensitive and resistant to this dose of Craviten, serum dopamine beta-hydroxylase activity was initially twice as high as that in healthy controls. After Craviten administration, enzyme activity normalized in the sensitive persons only, parallelly with rhythm normalization. In this group of patients the initially increased erythrocyte membrane ATPase activities (total and ouabain-insensitive) also normalized. PMID- 3024907 TI - Biochemical markers in sarcoidosis. AB - The clinical course of sarcoidosis is varying and unpredictable. Once the diagnosis has been made, the clinician needs simple tests to detect and predict remission or progression, to determine whether treatment is effective or not, and to assess the clinical activity of the disease. Sarcoidosis is a multisystem disease, but the lungs are almost always involved. Traditionally, the clinical management has therefore included chest X-rays and lung function studies. Extrapulmonary lesions have been followed in different ways. Sensitive and reproducible biochemical tests would be helpful in evaluating the clinical course of patients with sarcoidosis, if they measure functions related to the granulomatous inflammation. This review will deal with measurements of serum and urinary calcium, and 1,25-dihydroxyvitamin D. The usefulness of single and serial determinations of lysozyme, angiotensin converting enzyme, beta 2-microglobulin, collagenase, carboxypeptidase and glucuronidase in serum, bronchoalveolar lavage fluid, and other biological fluids will be discussed. PMID- 3024908 TI - Cytomegalovirus infection: a pediatrician's perspective. PMID- 3024909 TI - Production of superoxide by phagocytic leukocytes: a paradigm for stimulus response phenomena. PMID- 3024911 TI - Possible role of immunoglobulin recombination sequences in the genesis of variant t(2;8) translocations of Burkitt lymphoma. PMID- 3024910 TI - Regulation of adrenergic receptor function by phosphorylation. AB - Mounting evidence suggests that the physiological function of the various subtypes of adrenergic receptors is controlled by phosphorylation/dephosphorylation reactions. It seems intuitively unlikely that this phenomenon will be limited simply to the adrenergic receptors, since these receptors share transmembrane signaling pathways with a host of other plasma membrane receptors. Different types of kinases appear to be involved. On the one hand, phosphorylation reactions may operate in a classical feedback regulatory sense. Thus, the cAMP-dependent protein kinase, once activated by a beta-agonist, can feedback-regulate the function of the receptors by phosphorylating and desensitizing them. Similarly, protein kinase C appears to be able to feedback regulate the function of alpha 1-adrenergic receptors by phosphorylation. There may also be "cross talk" between the systems. Thus, protein kinase C, when stimulated by phorbols, is able to phosphorylate and desensitize the beta adrenergic receptors. Moreover, very recently we have found that the cAMP dependent protein kinase can phosphorylate the alpha 1-adrenergic receptors in vitro. These are examples of one transmembrane signaling system regulating the function of another. Perhaps most interestingly, it appears that there may be a previously unappreciated class of receptor kinases in the cytosol of cells. The first of these, which we have recently found and named beta-ARK, serves to phosphorylate only the agonist-occupied form of the beta-adrenergic receptor. As noted, it is somewhat analogous to the rhodopsin kinase. Such highly specific receptor kinases, which can phosphorylate only the agonist-occupied form of a receptor, represent a potentially elegant mechanism for controlling the function of receptors in a fashion which is linked to their physiological stimulation. How widespread such kinases are, and the actual roles which they play in regulating receptor function, remain to be determined. Finally, it should be stressed that although this review has focused on the regulatory role of receptor phosphorylation, it is by no means our intent to suggest that receptors are the only locus for physiological control of sensitivity to hormone and drug reaction. There is already evidence that guanine nucleotide regulatory proteins can be regulated, and it seems likely that each of the components of the system, including the adenylate cyclase, are likely to be involved in various forms of complex regulation. To date, however, the receptors represent that component of the system whose regulation we understand in the greatest detail. PMID- 3024912 TI - Tumors of newborn NFS/N mice infected with murine retroviruses containing avian v myc. PMID- 3024913 TI - Normal and neoplastic B cell development in the bursa of fabricius. PMID- 3024915 TI - Molecular analysis of myc gene mutants. PMID- 3024914 TI - Restrictions that influence avian leukosis virus-induced lymphoid leukosis. PMID- 3024916 TI - Induction of fgr proto-oncogene mRNA in B lymphocytes as a consequence of Epstein Barr virus infection. PMID- 3024917 TI - Epstein-Barr virus induced differentiation of early B-lineage cells. PMID- 3024918 TI - Epstein-Barr virus gene expression during primary B-lymphocyte infection, in transformed and Burkitt lymphoma-derived cell lines. PMID- 3024919 TI - EBV-activation of human B-lymphocytes. PMID- 3024920 TI - Induction of hematopoietic tumors using a viral construct containing c-myc cDNA from normal mouse spleen. PMID- 3024921 TI - Altered c-myc RNA metabolism in Burkitt's lymphomas and mouse plasmacytomas. PMID- 3024922 TI - Studies on c-myc regulation in normal and transformed cells. PMID- 3024924 TI - Determinants of Abelson murine leukemia virus pathogenesis. PMID- 3024923 TI - Transformation and insertional mutagenesis in vitro of primary hematopoietic stem cell cultures. PMID- 3024925 TI - Lymphohematopoietic and other malignant neoplasms occurring spontaneously in transgenic mice carrying and expressing MTV/myc fusion genes. PMID- 3024926 TI - Transposable elements and cancer. PMID- 3024927 TI - Exploratory nail plate removal as a diagnostic aid in painful subungual tumors: glomus tumor, neurofibroma, and squamous cell carcinoma. AB - Three rare, painful subungual tumors were precisely identified and evaluated following exploratory nail plate removal. Surgical excision of each was curative. PMID- 3024928 TI - Angiotensin converting enzyme in bronchoalveolar lavage in ARDS. AB - Angiotensin converting enzyme (ACE) is present in the endothelial cells of the normal lung where it converts angiotensin I to angiotensin II and inactivates bradykinin. It has been suggested that during endothelial injury ACE is sloughed into the blood, and that if the alveolar capillary membrane is injured, also into the alveolar lining fluid. Seven patients with adult respiratory distress syndrome (ARDS), were compared to 11 normal control subjects, nine patients with sarcoidosis, and six with idiopathic pulmonary fibrosis. Total, differential cell counts and ACE determinations were performed on bronchoalveolar lavage fluid in the ARDS group. ACE was detectable in the BAL of all but one ARDS patient. It was concluded that BAL ACE is elevated in some ARDS patients, especially those with infectious causes of lung injury. Increased ACE may reflect endothelial damage or local increase in ACE production in response to sepsis. PMID- 3024929 TI - Results of an analysis of the subcellular structure in 20 cases of primary hepatocellular carcinoma. PMID- 3024930 TI - [Phylloides tumors of the breast. A rare disease picture with a predominantly favorable prognosis]. AB - The phyllodes tumours, a group of rare (0.3%) neoplasms of the breast, consisting predominantly of mesenchymal tissue, are presented by an own case. A certain distinction between benign and malignant types can not always be obtained. The tumours are characterized by a high rate of local recurrences, but their prognosis is better than that of breast carcinomas, even in case of malignity. Metastasizing tumours mostly spread by an hematogenic pathway. The therapeutic access is surgical by complete local excision with a margin of security. Other oncological therapies failed to be successful. PMID- 3024932 TI - [Advances in the research on vitamin D and its metabolites]. PMID- 3024931 TI - Organization and chromosomal distribution of a novel repetitive DNA component from Muntiacus muntjak vaginalis with a repeat length of more than 40 kb. AB - The organization and chromosomal distribution of the repetitive DNA component IB from Muntiacus muntjak vaginalis (MMV) was investigated. DNA fragments of component IB were cloned in cosmids and their structure analysed using restriction nucleases and blot-hybridization experiments. Two cosmids were found to be practically identical by restriction enzyme mapping. The repeat unit of component IB DNA is more than 40 kb and contains the 11 and 18 kb Bam HI fragments, which have previously been shown to cross-hybridize with MMV satellite IA. In addition, the repeat unit contains long stretches of DNA sequences which are unique to component IB. In situ hybridization experiments showed that component IB has the properties characteristic of long interspersed repetitive DNA rather than tandemly repeated satellite DNA. Consistent with this conclusion, only a minor fraction of component IB is located on the X chromosome as demonstrated by the analysis of somatic cell hybrids. This is in marked contrast to satellite IA that is specific for the X chromosome. These results have interesting implications for the evolution of the component I DNA family of the MMV genome. PMID- 3024934 TI - Cronkhite-Canada syndrome. PMID- 3024933 TI - Histopathology of colorectal carcinomas and adenomas in cancer family syndrome. AB - Seventy-five colorectal carcinoma patients (100 separate cancers) with verified cancer family syndrome were re-examined for the evaluation of histologic characteristics in carcinomas and adenomatous polyps in this inherited syndrome in a comparison with control patients with colorectal carcinoma but no hereditary background. In the cancer family syndrome group there were significantly more mucinous carcinomas (35 to 39 percent vs. 20 percent; P less than 0.05-0.01), and also more poorly differentiated tumors (24 vs. 12 percent) than in the control group. The differences could not be explained by the site or stage of the tumors or by the age or sex of the patients. Additional adenomas occurred quite often both in cancer family syndrome patients (19 percent) and in the controls (16 percent). In the cancer family syndrome group, however, there were more adenomas with moderate or severe dysplasia (P less than 0.01) and more adenomas with villous features (P less than 0.05) than in the control group. Mucinous histologic features in colorectal carcinoma, although not fully specific, might be characteristic of cancer family syndrome, and thus serve as one sign in the identification of the syndrome. The presence of the adenoma-carcinoma sequence in cancer family syndrome also was supported, and the histologic aggressivity of the associated adenomas might signify an accelerated advancement of this phenomenon in cancer family syndrome. PMID- 3024935 TI - Cytopathology of fine-needle aspiration biopsy of the brain and spinal cord. AB - The cytomorphologic findings of 37 intraoperative fine-needle aspiration (FNA) biopsies are presented. Thirty-two of the biopsies were performed at the time of craniotomy, and five were performed through a burr-hole. All cases used direct smear preparations stained with Papanicolaou and modified Wright stain (Diff Quik). Immediate evaluation of the Diff-Quik-stained smears was performed for assessment of the adequacy of the specimen, and in most cases, a rapid preliminary diagnosis (similar to a frozen-section report) was rendered in the operating room. Cytopathologic features of anaplastic astrocytomas, glioblastoma multiforme, pituitary adenoma, meningioma, metastatic carcinoma, epidermoid cyst, herniated disc, hemangioblastoma, cerebral hemorrhage, and malignant lymphoma are described and illustrated. The cytologic preparation demonstrated superior cellular detail, which served as a considerable aid for diagnosis, and lacked the artifactural distortion often seen in the frozen-section preparations. Reliance on the fine-needle aspiration biopsy specimen enabled better triage of the small amount of tissue often available for permanent sections and special studies including electron microscopy and immunohistochemistry. Although not meant to replace tissue biopsy, FNA of the central nervous system though burr holes under radiologic guidance can be advantageous in selective cases. PMID- 3024936 TI - Immunocytochemical localization of human papilloma virus and cytomorphologic correlation in smears and biopsies of cervical flat condylomata. AB - The cytomorphologic features in cervical biopsies and smears associated with human papilloma virus antigen (Ag) expression as demonstrated by immunoperoxidase staining techniques are presented. There was good concordance between cytology and biopsy results with respect to immunoperoxidase staining for human papilloma virus Ag. Cytomorphologic scoring of low-grade lesions (cervical intraepithelial neoplasias, grades O and I) showed atypical mitoses, macronucleated koilocytes, superficial cell keratohyaline granules, and multinucleation to be more frequent in Ag+ biopsy specimens. Chronic inflammation was more evident in Ag- biopsy specimens (P less than 0.05). Parakeratosis was more frequent in Ag+ cytologic specimens. Koilocytosis was not a reliable indicator of Ag expression in either preparation. Human papilloma virus (HPV) Ag testing by immunoperoxidase techniques appears to be a useful adjunct for screening low-grade atypias of the cervix. PMID- 3024937 TI - Cytomegalovirus infection of the lung: cytomorphologic diagnosis by fine-needle aspiration cytology. AB - Cytomegalovirus (CMV) infection of the lung was diagnosed by fine-needle aspiration cytology (FNAC) in a patient who underwent renal transplantation due to end-stage renal failure. This case illustrates that CMV can be diagnosed by FNAC and, when seen, must be reported in an immunocompromised host. PMID- 3024938 TI - Carcinoid tumors of the lung: cytologic differential diagnosis in fine-needle aspirates. AB - Pulmonary carcinoid tumors presenting as peripheral or coin lesions are rare and radiologically may resemble other primary or metastatic neoplasms in the lung. This study consisted of the cytologic evaluation of fine-needle aspirates from five peripheral carcinoid tumors of the lung with particular reference to the differences between the cytologic manifestations of this neoplasm and of small cell carcinoma and well-differentiated adenocarcinoma. Aspirates of typical carcinoid tumors are characterized by isolated cells and loose aggregates of cells; spindle- and oval-shaped cells of uniform size with scanty, pale eosinophilic cytoplasm; and nuclei with evenly dispersed finely granular chromatin and usually prominent single micronucleoli with occasional macronucleoli. In contrast, the cells of small-cell carcinoma are pleomorphic and arranged in noncohesive loose aggregates, their cytoplasm is scanty, and they show nuclear molding. Their nuclei are hyperchromatic with fine to coarsely granular chromatin. The cells of well-differentiated adenocarcinoma are arranged in three-dimensional clusters or loose aggregates of relatively uniform cells with a columnar configuration. The nuclei are uniformly round and hyperchromatic with finely granular chromatin. Macronucleoli are usually prominent. This study demonstrates that cytologic analysis of fine-needle aspirates can play a significant role in the evaluation and management of peripheral lung tumors. PMID- 3024939 TI - [Inhibitory effect of tumor promoters on the intercellular metabolism of lucifer yellow in a culture of transformed Djungarian hamster fibroblasts]. PMID- 3024940 TI - [Synergistic effect of stimulators of the adenylate cyclase system and phosphoinositide metabolism in the perfused rabbit aorta]. PMID- 3024941 TI - [The rat genome contains an active locus structurally associated with the mos proto-oncogene]. PMID- 3024942 TI - [Reactivation of DNA synthesis in macrophage heterokaryons and cells transformed with ts-mutant virus SV40 is temperature-dependent]. PMID- 3024943 TI - [Mechanism of the antioxidant effect of ceruloplasmin]. PMID- 3024944 TI - [Differences in opioid binding in the left and right visual cortex of the turtle]. PMID- 3024945 TI - Selective drug action on two Ca2+ uptake processes in normal and hyperexcitable presynaptic membrane. AB - Synaptic transmission in the bullfrog sympathetic ganglion in vitro reflects Ca2+ dependent presynaptic and Na+-dependent postsynaptic membrane excitabilities. Aminopyridines (e.g., 3-AP, 4-AP,3,4-DAP) produce Ca2+-dependent presynaptic membrane hyperexcitability (increased transmitter release), evident as synchronized, stimulus-bound repetitive postsynaptic spike responses (SBR) to each single preganglionic stimulus (0.1 Hz). The SBR induced by 3,4-DAP is selectively eliminated as the normal [Ca2+]0 of 1.8 mM is reduced to 0.6 mM, whereas failure of the primary spike response begins only at 0.33 mM [Ca2+]0 and is complete at 0.07 mM. These differences in Ca2+ dependence suggest that two separate presynaptic Ca2+ uptake processes are involved in transmission (the primary spike) and in SBR. The authors' present work with Ca2+-channel blockers (CCBs) reinforces the preceding evidence for quantitatively separate presynaptic Ca2+ uptake processes. Thus, aminopyridine-induced SBR is also selectively abolished by verapamil (V) or diltiazem (D). From threshold to complete abolition of SBR the effective CCB ranges are: V,0.04-0.15 mM; D,0.01-0.08 mM. Higher concentrations are required to block synaptic transmission (the primary spike), the effective CCB ranges being: V,0.25-0.75 mM; D,0.1-0.4 mM. The CCBs thus display considerable concentration selectivity in stabilizing the hyperexcitable presynaptic membrane and in depressing its normal excitability. This is fully analogous to the authors' earlier work, in which SBR induced by physostigmine or 3,4-DAP was selectively abolished by d-tubocurarine or lidocaine concentrations below transmission-blocking levels.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3024946 TI - A pH-dependent disordering efficacy of new local anaesthetics with carbanilate structure. A spin label study in model membranes. AB - Electrophysiological investigations of new drugs with carbanilate structure (heptacaine, carbisocaine) on axons and sciatic nerves of the rat have shown their high local anaesthetic effect at physiological pH. This effect is not only maintained at acid pH (6.0); it even increases on shifting pH from basic to acid values. To study this unique pH dependence of the effect of new local anaesthetics, drug-lipid interactions were investigated in liposomes prepared from egg yolk phosphatidylcholine or dimyristoyl phosphatidylcholine. The effect of the local anaesthetics on the molecular ordering of lipids in the liposomes was measured from spectral anisotropy of spin-labelled fatty acid incorporated in the membrane. It was concluded that the change of pH from 9.5 to 6.5 not only sustains the disordering effect of heptacaine and carbisocaine; it even potentiates it with the maximum disordering effect recorded at pH 6.5. This trend towards pH dependence in the effect of the new local anaesthetics is similar to that observed in biological preparations and is quite unlike the case of classic local anaesthetics (procaine, lidocaine). The possible mechanism of pH dependence is discussed. PMID- 3024947 TI - [Estrogen receptor status in breast carcinoma. Results of comparative immunohistochemical and biochemical studies]. AB - Comparative biochemical and immunohistochemical investigations on receptors were performed on 100 breast cancers of different types. The biochemical analysis was carried out by means of radioimmunoassay (RIA) and immunohistochemical detection was performed with monoclonal antibodies against nuclear receptor protein (ER ICA, Abbott Lab.). The semiquantitative evaluation is based on a histological score considering a graduated staining intensity and the percentage of positively reacting cells. The results of the two methods corresponded in 92% in positive and in 68% in negative receptor detection. The age-dependence is confirmed. The nuclear grading and histological grading show that the number of estrogen receptors gradually decreases with increasing degree of malignancy of the carcinomas. The advantage of immunohistochemical receptor assay consists in the microscopic investigation of homogeneous and heterogeneous degrees of intensity as well as in the possibility of determining receptor status in very small breast cancers, in punch biopsies and in tumours in which the tumour parenchyma is largely necrotic, sclerosed or surrounded by large amounts of mucus. PMID- 3024948 TI - [Therapy of primary resistant or recurrent small cell bronchial carcinoma with vindesine and ifosfamide]. AB - A combination of vindesine (3 mg/m2, day 1) and ifosfamide (60 mg/kg, days 1-5 + Mesna) was administered every three weeks to 11 patients with primary resistant and 23 with recurrent small-cell bronchial carcinoma. All patients had been pre treated with chemotherapy, 16 in addition with radiotherapy. At the onset of the vindesine-ifosfamide treatment the cancer was in a localized regional stage in ten patients, while in 24 it was in a more widely spread stage. In 29 patients whose treatment results could be evaluated the remission rate was 38%, with two complete and nine partial remissions. In a further eight patients the cancer was arrested. The patients with complete remission (for 46 and 53 weeks, respectively), those with partial remission (median of 39 weeks) and those with stationary disease (median of 31 weeks) survived significantly longer than those with progressing disease (13 weeks). There was no correlation between treatment result and pre-treatment. On recurrence after complete remission or in the localized regional stage the remission rate was 70% and 60%, respectively, and the survival time was extended in 90% of cases. In addition to nausea, alopecia and myelosuppression, side-effects included vomiting, reversible CNS symptoms, polyneuropathy and urotoxicity. On the basis of acceptable toxicity, combined vindesine and ifosfamide constitute an effective treatment of otherwise treatment refractory cases of small-cell bronchial carcinoma. PMID- 3024951 TI - Use of technetium--99m uptake studies in the rapid assessment of thyroid function. PMID- 3024950 TI - Sorbitol, myo-inositol and sodium-potassium ATPase in diabetic peripheral nerve. AB - Slowing of nerve conduction, a hallmark of both experimental and human diabetic neuropathy, is improved or corrected by aldose reductase inhibitors such as sorbinil. Animal experiments suggest that a myo-inositol-related defect in nerve sodium-potassium adenosine triphosphatase (ATPase) is responsible for the acute reversible slowing of nerve conduction in diabetes mellitus. This myo-inositol related defect is at present viewed as a cyclic metabolic defect. Aldose reductase inhibitors have been shown to restore to normal both the myo-inositol content and the sodium-potassium ATPase activity of nerve. This suggests that the acute effects of aldose-reductase inhibitors on nerve conduction in both diabetic animals and human patients may be modified by the correction of an underlying myo inositol-related defect of nerve sodium-potassium ATPase. Furthermore, this myo inositol-related defect may contribute to other biochemical, functional and structural abnormalities of diabetic peripheral neuropathy. PMID- 3024952 TI - Comparative kinetics study of the evolution of freshwater aquatic toxicity and biodegradability of linear and branched alkylbenzene sulfonates. AB - Evolution of both primary biodegradability and acute toxicity to daphnia and zebra fish of a linear alkylbenzene sodium sulfonate (LAS) and a branched alkylbenzene sodium sulfonate (BAS) have been measured simultaneously. In six of eight experiments, LAS was biodegraded to 90% in 7 days and BAS to 70% in 7 days. In the two other experiments, both LAS and BAS have shown the same biodegradation speed and reached the same biodegradation level in 7 days: 45% in one experiment and 55% in the other. The composition of bacteria population and the level of cellular ATP of the inoculum play a decisive role in the biodegradation. These results confirm that it is essential to know the composition of bacteria population present in the inoculum as well as their biochemical characteristics to accurately interpret results of biodegradation tests. In the case of a rapid primary biodegradation of LAS and BAS, the acute toxicity of LAS remains three times as high as that of BAS for at least 24 hr toward daphnia and 48 hr toward zebra fish. Their acute toxicity to daphnia and zebra fish become equivalent only after 72 hr. When primary biodegradation of both products is slower, the acute toxicity of LAS remains higher than that of BAS for more than 7 days. PMID- 3024953 TI - Effect of morphine on hypothalamic corticotropin-releasing factor (CRF) and pituitary-adrenocortical activity. AB - The effects of intraperitoneal and intra-third ventricular administration of morphine on the hypothalamic corticotropin-releasing factor (CRF) and the pituitary-adrenocortical activity were examined in unanesthetized, freely moving rats. Hypothalamic CRF was measured by rat CRF radioimmunoassay. Intraperitoneal or intra-third ventricular administration of morphine increased blood concentrations of ACTH and corticosterone while intraperitoneal administration tended to increase CRF concentration in the whole hypothalamus including the median eminence and intra-third ventricular administration increased CRF concentration in the hypothalamus excluding the median eminence. However, morphine seemed to inhibit the increase in CRF concentration in the hypothalamus induced by the ether-laparotomy stress. The main site of morphine action on the hypothalamo-pituitary-adrenocortical system seemed to be in the hypothalamic area. PMID- 3024954 TI - Cortisol responsiveness to insulin-induced hypoglycemia in Cushing's syndrome with huge nodular adrenocortical hyperplasia. AB - A 51-yr-old male patient with a 3 yr history of Cushing's syndrome is described. The baseline plasma cortisol level was elevated, while the plasma ACTH levels remained at an undetectable level. Dynamic testing of pituitary-adrenal function revealed no suppression after 8 mg of dexamethasone, and there was no response to metyrapone or CRF, while plasma cortisol showed a hyperresponse to synthetic ACTH. Plasma cortisol responded to insulin-induced hypoglycemia without an obvious ACTH response. These and the computerized tomography data suggested a "huge" bilateral nodular adrenocortical hyperplasia which was later confirmed by surgery. The left and right adrenal glands weighed 55 and 76 g, respectively. In vitro experiments, using the adrenal tissue, showed that there was an adrenal cortisol response to 1-39 ACTH but not to regular insulin, arginine vasopressin, angiotensin II, norepinephrine or epinephrine. These results indicate that plasma cortisol responded to a slight hypoglycemia-induced plasma ACTH change which was not detected in the ACTH radioimmunoassay or to factors other than ACTH which might be induced by hypoglycemia. PMID- 3024955 TI - Demineralized dentin, hydroxylapatite and dentin chips as apical plugs. PMID- 3024956 TI - Gastroduodenal polyps in familial adenomatous and juvenile polyposis. AB - Upper gastrointestinal endoscopy revealed gastroduodenal polyps in 45 (90%) out of 50 patients with familial adenomatosis (FA), and in 11 (92%) out of 12 patients with juvenile polyposis (JP). The polyps in the fundic and corpus regions of the stomach were hamartomatous cystic body gland polyps in 28 (56%), and adenomas in 3 (6%) of the FA patients, while 6 (50%) JP patients had hyperplastic polyps in the corresponding region. The polyps in the gastric antrum and duodenum were adenomas in 6 (12%) and 25 (50%) of the FA patients, respectively, while the polyps of the corresponding regions were hyperplastic in 10 (83%) and 2 (17%) of the JP subjects. One focal adenomatous lesion in an antral hyperplastic polyp, and one duodenal adenoma were found in the JP patients. Furthermore, one duodenal adenocarcinoma was observed among the FA subjects. It is concluded that gastroduodenal polyps belong inherently both to adenomatous and to juvenile varieties of gastrointestinal polyposis, and that these two conditions have characteristic histological patterns and topography of polyps in the upper gastrointestinal tract. PMID- 3024957 TI - Decreased GABA, benzodiazepine, and picrotoxinin receptor binding in brains of rats after cobalt-induced epilepsy. AB - In previous studies of experimental and human epilepsy, defects have been shown in the gamma-aminobutyric acid (GABA) receptors. We further investigated the role of the GABA/benzodiazepine/picrotoxinin receptor complex in the epileptic focus and also in other regions of the rat brain. The focus was induced by cobalt implantation to the right motor cortex, and the brains were dissected 16-19 days after the operation. Benzodiazepine (using [3H]flunitrazepam as a ligand; FLU), GABA [3H]muscimol; MUS), and picrotoxinin [( 35S]t-butylbicyclophosphorothionate; TBPS) receptor bindings were measured in different brain areas and the values were compared with glass-implanted controls. In the focal area, the specific receptor binding decreased in the order TBPS greater than FLU greater than MUS. In the perifocal area only TBPS binding decreased, and Scatchard analysis showed a decrease in the number of binding sites (p less than 0.05) without any effect on binding affinity. No change was seen in the binding characteristics of the other areas studied. According to our results, in cobalt-induced epilepsy the GABA/benzodiazepine/picrotoxinin receptor complex is modulated in the focal area; this may lead to a defect in chloride conductance, which in turn induces disturbed control of neuronal activity in the epileptic focus. PMID- 3024958 TI - Failure of ACTH to alter transfer kindling in the immature brain. AB - For a study of the effects of ACTH on established seizures, 16-day-old rats that had been previously kindled in the amygdala were given a high daily dosage of ACTH gel for 15 days and then underwent transfer kindling in the contralateral amygdala. There were no significant differences in the rate of transfer kindling between the controls and ACTH-treated rats. This study indicates that in the immature animal ACTH does not alter the kindled state. PMID- 3024949 TI - The treatment of heart failure. A methodological review of the literature. AB - In this article literature concerning the major drugs used in the treatment of heart failure is reviewed. Because of major discrepancies in results from short term and uncontrolled studies versus long term randomised control trials, only the latter group of studies are addressed in detail. Of 3 randomised control trials of digoxin, 1 has been positive, and 2 negative. Digoxin is probably of benefit to a minority of heart failure patients. Four randomised control trials of oral nitrates have shown a reduction in left ventricular filling pressure, and trends favouring active treatment for the indices of exercise capacity and functional status. Of 2 randomised control trials of hydralazine one is totally negative, the other difficult to interpret because of major loss of patients to followup. Of 5 trials of quinazolone derivatives (prazosin and trimazosin), 2 have been positive, 2 showed non-statistically significant trends favouring active treatment, and 1 was completely negative. These results are consistent with a modest benefit of prazosin and trimazosin in some heart failure patients. Five trials of angiotensin-converting enzyme inhibitors (captopril and enalapril) have shown dramatic and consistent benefit in exercise capacity and functional status. These results support a clinical policy of initial treatment with diuretics and addition of either captopril or enalapril for patients who remain symptomatic on optimal diuretic therapy. A trial of digoxin is warranted in patients whose functional capacity remains reduced on this regimen. PMID- 3024960 TI - Net-charge models in the analysis of ion and electron transfer in enzyme reactions. AB - A number of key questions arising in the study of proton-relay chains and of the driving forces causing them to operate are discussed with main emphasis on the net-charge distribution of relevant model systems. Special reference is made to the role of water, histidine imidazole, and peptide bonds and to the description of hydride transfer in ADH-NAD+ complexes. Suggestions are given for treating H bonds by the so-called Del Re method. PMID- 3024959 TI - Clinical effects of thyrotropin-releasing hormone for severe epilepsy in childhood: a comparative study with ACTH therapy. AB - We investigated the effects and side effects of thyrotropin-releasing hormone (TRH) on severely epileptic children to evaluate the clinical usefulness of TRH in the treatment of epilepsy and compared them with the results of ACTH therapy. The subjects were 64 patients admitted consecutively between 1980 and 1986. Their seizures were frequent, more than one a day or more than one a week. The subjects were divided into two groups; 33 patients treated with ACTH and 31 treated with TRH. The mean follow-up periods in TRH and ACTH therapy were 8 months and 3.0 years, respectively. The daily dose of TRH-t 0.5-1 mg was administered intravenously (i.v.) or intramuscularly (i.m.) for 1-4 weeks. The follow-up periods were 3-12 months (mean 6 months). In the TRH group, complete control of seizures was achieved in 7 of 13 (53.7%) of those with infantile spasms, and marked improvement of EEGs were observed in 8 of 13 (61.5%) of them. In the ACTH group, seizure cessation was observed in 75% of infantile spasms. Of the patients who received ACTH, 66.7% had various side effects, including pneumonia, huge subcutaneous abscess, hypokalemia, cataracts, and brain shrinkage as shown on computed tomography (CT), whereas only 16.7% of the patients treated with TRH had transient reduction of urine volume without other laboratory and physical abnormalities. The results of the study indicated that some patients who received TRH had cessation of infantile spasms and improved EEG findings with no serious side effect. Because of the untoward side effects of ACTH therapy, TRH is considered a possible new treatment for children with infantile spasms. PMID- 3024961 TI - Effect of local environment and protein on the mechanism of action of superoxide dismutase. AB - Quantum mechanical simulations of the mechanism of action of superoxide dismutase (SOD) indicate that the presence of Arg-141 in the active site of the enzyme is responsible for the formation of an intermediate complex between superoxide and the enzyme in which the copper is not reduced. The analysis of the local environmental effects of Arg-141 shows that this residue prevents the reduction of copper by forming a hydrogen bond to superoxide and by generating an electric field in the active site that opposes the transfer of an electron from superoxide to copper. The protein enhances the effect of the opposing field generated by Arg 141. Local changes in the environment of the copper ion, simulated by stretching the Cu-NE2 (His-61) bond also do not induce an electron transfer from superoxide to copper. The protein increases the energies required for this stretch through the electric field it generates near the active site. These results are further support for the new proposed mechanism of action of SOD which is based on the inability of superoxide to reduce the cupric ion in the enzyme. PMID- 3024962 TI - Mapping the collagen-binding site of human fibronectin by expression in Escherichia coli. AB - The collagen-binding domain of human fibronectin has been expressed as a cro/beta galactosidase fusion protein in Escherichia coli. The hybrid polypeptide was recognized by an anti-(human plasma fibronectin) serum and bound specifically to gelatin-Sepharose. The collagen-binding region was subdivided by constructing a series of overlapping bacterial expression plasmids. The fusion proteins produced by these constructs were analysed for gelatin-binding activity. The results indicate that the binding site lies within an approximately 12.5 kd fragment of fibronectin, and show that the following 14 amino acid sequence is critical for gelatin-binding activity: Ala-Ala-His-Glu-Glu-Ile-Cys-Thr-Thr-Asn-Glu-Gly-Val Met. This sequence links the second type II homology unit with the adjacent type I repeat in the amino-terminal third of the fibronectin molecule. PMID- 3024963 TI - Mutations blocking the transport of the influenza virus hemagglutinin between the rough endoplasmic reticulum and the Golgi apparatus. AB - Mutants ts1 and ts227 of fowl plague virus have a temperature-sensitive defect in the transport of the hemagglutinin from the rough endoplasmic reticulum to the Golgi apparatus. The primary structure of the hemagglutinin of the mutants and of a number of revertants derived from them has been analysed by nucleotide sequencing. The transport block of the hemagglutinin of ts227 can be attributed to a single amino acid exchange. It involves the replacement of aspartic acid at position 457 by asparagine thereby introducing a new glycosylation site which appears to be located in a cryptic position in the lower part of the hemagglutinin stalk. Attachment of carbohydrate to this site is temperature dependent. At permissive temperature only a small fraction of the monomers (approximately 30%) is glycosylated in this position, whereas at nonpermissive temperature this is the case with all subunits. The data suggest that under the latter conditions the new oligosaccharide interferes by steric hindrance with the trimerization of the hemagglutinin. The hemagglutinin of ts1 has an essential amino acid exchange at position 275 where serine is replaced by glycine. This substitution may increase the flexibility of the molecule in the hinge region between the globular domain and the stalk. The exchange of a conserved glutamic acid residue at position 398 that is involved in the interaction between different monomers contributes also to the structural instability of the ts1 hemagglutinin. These observations support the notion that the transport of the hemagglutinin from the rough endoplasmic reticulum to the Golgi apparatus depends on trimer assembly. PMID- 3024964 TI - Chromosome 8 breakpoint far 3' of the c-myc oncogene in a Burkitt's lymphoma 2;8 variant translocation is equivalent to the murine pvt-1 locus. AB - The 2;8 variant translocation of human Burkitt's lymphomas is closely related cytogenetically to the t(6;15) of murine plasmacytomas; both involve a reciprocal exchange between the Ig kappa locus and a band region indistinguishable from that bearing the c-myc oncogene. To define their molecular relationship, we have compared cloned chromosome 8 DNA from the t(2;8) breakpoint in the human Burkitt's lymphoma JBL2 with cloned DNA from the murine pvt-1 locus, the major chromosome 15 breakpoint region in murine t(6;15). DNA sequencing and Southern blot analysis shows that these two regions are homologous. Thus the t(2;8) in JBL2 is the molecular equivalent of many murine t(6;15). The murine pvt-1 locus lies an unknown distance 3' of c-myc; analysis of DNA from several tumours with c myc amplification reveals that pvt-1 is co-amplified in at least one case, placing pvt-1 approximately 100-500 kb 3' of c-myc. The significance of these results with respect to the role of pvt-1 in tumorigenesis is discussed. PMID- 3024965 TI - Intragenic pausing and anti-sense transcription within the murine c-myc locus. AB - We present a detailed analysis of strand-specific transcription in different regions of the murine c-myc locus. In normal and transformed cell lines, RNA polymerase II directed transcription occurs in the sense and anti-sense direction. Three noncontiguous regions show a high level of transcription in the anti-sense orientation: upstream of the first exon, within the first intron and in the 3' part of the gene (intron 2 and exon 3). In a cell line carrying a c-myc amplification (54c12), anti-sense transcription is not uniformly increased throughout the locus and is differentially affected by inhibition of protein synthesis. These results suggest that anti-sense transcription in various parts of the locus is independently regulated. In the sense orientation, transcriptional activity is higher in the first exon than in the rest of the gene indicating that transcription pauses near the 3' end of the first exon. The extent of this intragenic pausing varies among different cell lines and is most severe in cells with a c-myc amplification. Transcription initiation and pausing are both negatively regulated by labile proteins. PMID- 3024966 TI - Induction of interleukin 2 receptor gene expression by p40x encoded by human T cell leukemia virus type 1. AB - Human T-cell leukemia virus type 1 (HTLV-1) is an etiologic agent of adult T-cell leukemia (ATL). A viral product, p40x, encoded by the pX sequence of HTLV-1 is a trans-acting transcriptional activator of the long terminal repeat (LTR) and has been suspected of involvement in leukemogenesis, activating the cellular genes. The cellular interleukin-2 (IL-2) and its receptor (IL-2R), the latter of which is expressed on ATL leukemic cells, were shown to be transiently induced by transfection of plasmid pMTPX expressing pX in two T-cell lines, Jurkat and HSB 2, but not in other human T- or B-cell lines. The cell type specificity of IL-2R induction by pX expression was the same as that by phytohaemagglutinin/phorbol ester activation, indicating the requirement for some specific cellular factors or a certain state of cellular differentiation. Induction of IL-2 and IL-2R at mRNA level was also demonstrated in transfected cells. Transfections with mutants of pMTPX in which the open reading frames for p40x, p27x-III and p21x-III were inactivated indicated that p40x alone was sufficient for induction of the IL-2R in inducible cells. This induction of the IL-2R by p40x of HTLV-1 may contribute to preferential proliferation of HTLV-1 infected cells at an early stage of ATL development and eventually increase the number of putative target cells for malignant transformation. PMID- 3024967 TI - Sequences capable of restoring poly(A) site function define two distinct downstream elements. AB - Several recent studies have shown that a functional poly(A) site consists of both an AAUAAA element as well as sequences downstream of the cleavage site. Two downstream regions were analyzed in an attempt to accurately locate and define the critical sequences. Chemically synthesized oligonucleotides of sequence from the early SV40 and the adenovirus E2A poly(A) sites were able to restore efficient cleavage to a deleted SV40 poly(A) site. Inversion of the sequence completely abolished poly(A) site function. A series of base substitution mutants were generated in each downstream sequence. Certain single base changes drastically altered poly(A) site function. Thus, it is concluded that a defined downstream sequence of limited complexity is important for efficient processing of the primary transcript at the poly(A) site. The position of the downstream elements relative to the AAUAAA and cleavage site was found to be critical since moving either the E2 element or the SV40 element an additional 40 nucleotides downstream abolished function. There were differences, however, in the effect of spacing on the function of the two elements. This observation, along with the fact that the two sequences are clearly different, indicates that there are at least two distinct genetic elements that direct efficient cleavage at the poly(A) site. PMID- 3024968 TI - Alpha-thalassaemia caused by a poly(A) site mutation reveals that transcriptional termination is linked to 3' end processing in the human alpha 2 globin gene. AB - We have investigated the process of transcriptional termination in the duplicated human alpha globin genes which lie 4 kb apart on chromosome 16. In the human erythroleukemic cell line, K562, which expresses high levels of alpha globin, nuclear run-off experiments suggest that termination occurs within a region of 600 bp past the poly(A) site of both alpha 1 and alpha 2 globin genes. However, a thalassaemic alpha 2 globin gene with a non-functional poly(A) site AAUAAG, when transfected into HeLa cells, not only fails to 3' end process but also fails to terminate transcription. Studies on both steady-state RNA and nuclear run-off analysis of the primary transcripts show that transcription of the mutant alpha 2 globin gene reads through into the intergenic sequence past the normal termination site. These results demonstrate that transcriptional termination and 3' end processing of mRNA are coupled events for the alpha 2 globin gene. PMID- 3024969 TI - Formation of the 3' end on U snRNAs requires at least three sequence elements. AB - The structural requirements for 3' end formation on the Xenopus laevis U1B snRNA gene have been studied. Three sequence elements are shown to be required. The first is a conserved sequence element found immediately 3' of all vertebrate U snRNA genes studied so far. The second is a gene internal sequence potentially capable of forming a stem-loop structure close to the 3' end of the RNA. The third element lies upstream of these, and may be part of the gene promoter. Experiments designed to investigate the mechanism of 3' end formation on primary U1B snRNA transcripts failed to find evidence for a processing event. PMID- 3024970 TI - Isolation of the eclosion gene cluster and the developmental expression of the Gld gene in Drosophila melanogaster. AB - During the development of Drosophila melanogaster the expression of glucose dehydrogenase (GLD) changes from non sex-limited to male limited. We have isolated the Gld gene and three other functionally related genes in the eclosion gene cluster by the method of chromosome walking. The Gld gene has been identified by two deletions and a translocation which genetically define the gene. A 2.8-kb RNA has been identified as the putative GLD mRNA. The temporal and spatial expression of this RNA is correlated with the expression of the GLD enzyme and levels of the steroid hormone ecdysterone. Using single-strand antisense probes we have detected three RNA species. However these three transcripts are not derived from the Gld locus. One of these RNAs is weakly detected by the multiple cloning site of the pSP65 vector. The level of detection of this latter RNA is greatly increased by the insertion of a specific Gld gene fragment in the pSP65 vector. PMID- 3024971 TI - The Ace locus of Drosophila melanogaster: structural gene for acetylcholinesterase with an unusual 5' leader. AB - The Ace locus of Drosophila melanogaster has been mapped at the molecular level. cDNA clones from the locus have been isolated and their sequence determined, confirming that Ace forms the structural gene for acetylcholinesterase (AChE). The cDNAs have a 1950 nucleotide open reading frame from which the complete amino acid sequence of AChE has been deduced. The Drosophila enzyme is found to have extensive homology to the known sequence of Torpedo AChE. Ace cDNAs have an unusual structure with a long 5' leader and several short upstream open reading frames. PMID- 3024972 TI - Role of sog polypeptides specified by plasmid ColIb-P9 and their transfer between conjugating bacteria. AB - The sog gene of the conjugative plasmid ColIb-P9 specifies two sequence-related polypeptides with the N-terminal third of the larger product having DNA primase activity. To resolve the function of the C-terminal portion of the polypeptides, we constructed a ColIb mutant containing a Tn5 insertion in the 3' region of sog. The mutation truncated sog gene products without inactivating DNA primase and rendered the plasmid defective in conjugation. Tests for the presence of conjugative pili, for complementation by a sog+ recombinant, and for mobilization of small origin of transfer (oriT) recombinant plasmids indicated that the mutant ColIb allows conjugative aggregation of cells but it is defective in DNA transfer at some stage subsequent to its initiation at oriT. Physical evidence is given that normal sog polypeptides are among a group of proteins transferred selectively from the donor to the recipient cell by a conjugation-specific process. No transfer of the mutant sog proteins was detected. It is proposed that the C-terminal region of sog polypeptides facilitates transfer of single-stranded ColIb DNA between conjugating cells following initiation of transfer at the oriT site, and that in this role the proteins are transmitted to the recipient cell. PMID- 3024974 TI - In-vivo-modified gonococcal plasmid pJD1. A model system for analysis of restriction enzyme sensitivity to DNA modifications. AB - The 4207-bp cryptic plasmid (pJD1) of Neisseria gonorrhoeae has 5-methylcytosine bases present at several positions in the DNA sequence. Fortuitously, these modified bases lie in the recognition sequences of many restriction enzymes. This feature makes the cryptic plasmid a model system for assaying the effect of these modified cytosines on the activities of the following restriction endonucleases and their isoschizomers: R X AvaII, R X BamHI, R X BglI, R X Fnu4HI, R X HaeII, R X HaeIII, R X HhaI, R X HpaII, R X KpnI, R X MspI, R X NaeI, R X NarI, R X NciI, R X NgoI, R X NgoII, and R X Sau96I. Of particular interest was the finding that methylation of one of the external cytosines of the palindrome 5'-CCGG-3' prevented its cleavage by R X MspI, but not by R X HpaII as had been suggested by Walder et al. [J. Biol. Chem. (1983) 258, 1235-1241]. PMID- 3024973 TI - Identification of the human cytomegalovirus glycoprotein B gene and induction of neutralizing antibodies via its expression in recombinant vaccinia virus. AB - A human cytomegalovirus (HCMV) glycoprotein gene with homology to glycoprotein B (gB) of herpes simplex virus and Epstein-Barr virus and gpII of varicella zoster virus has been identified by nucleotide sequencing. The gene has been expressed in recombinant vaccinia virus and the gene product recognized by monoclonal antibodies and human immune sera. Rabbits immunized with the recombinant vaccinia virus produced antibodies that immunoprecipitate gB from HCMV-infected cells and neutralize HCMV infectivity in vitro. These data demonstrate a role for this protein in future HCMV vaccines. PMID- 3024975 TI - Purification and properties of guanylate kinase from bovine retinas and rod outer segments. AB - The presence of three soluble nucleotide phosphotransferases in bovine rod outer segments was demonstrated: guanylate kinase (EC 2.7.4.8), nucleoside-diphosphate kinase (EC 2.7.4.6) and adenylate kinase (EC 2.7.4.3). The enzyme guanylate kinase, which catalyzes the reaction GMP + ATP in equilibrium GDP + ADP, was purified to homogeneity from isolated bovine rod outer segments as well as from bovine retinas. The enzyme preparations obtained from both sources are identical in their chromatographic properties, molecular mass (20-23 kDa for both native enzyme and dodecylsulfate-denatured polypeptide), Km values (13 microM for GMP and 430 microM for ATP), specific activities, and nucleotide specificities. The enzyme's turnover number was estimated to be 130 s-1. The minimum amount of enzyme found in rod outer segments is about 1 copy per 800 rhodopsin molecules. The role of the enzyme in the cyclic GMP cycle in rod outer segments is discussed. PMID- 3024976 TI - Core region of Citrobacter lipopolysaccharide from strain PCM 1487. Structure elucidation by two-dimensional 1H-NMR spectroscopy at 500 MHz and methylation analysis/mass spectrometry. AB - The core structure of Citrobacter PCM 1487 lipopolysaccharide has been established using methylation analysis/mass spectrometry, chemical degradations and one- and two-dimensional 1H-NMR spectroscopy at 500 MHz. 1H-NMR assignments are given for all sugar components of the core oligosaccharide. In the formula shown below, the alternative locations of branch terminal heptose (LDHep) and diphosphorylethanolamine (PPEtN) residues are marked by dashed lines; dOclA stands for 3-deoxy-D-manno-octulosonic acid. (Formula: see text). The sample of the core oligosaccharide showed some microheterogeneity due to a slightly incomplete substitution by terminal N-acetylgalactosamine and a partial splitting of diphosphorylethanolamine residues. PMID- 3024977 TI - Conformational studies of d(m5CpGpm5CpG) and d(CpGpCpG) by 1H and 31P NMR. AB - Exhaustive conformational studies of d(CpG)2 and d(m5CpG)2, two convenient targets for DNA bisintercalating drugs, have been carried out by 1H and 31P NMR in low salt concentration and in the presence of 30% ethanol. Unambiguous 31P assignments of the B for are obtained with low-power heteronuclear decoupling experiments, while 31P assignments in the Z form are obtained by two-dimensional homonuclear chemical exchange experiments. The 31P chemical shifts and 3JH3'P coupling constants studied at various temperatures in methylated and non methylated tetranucleotides, are interpreted as resulting from conformational differences between the compounds. These features are corroborated by homonuclear proton nuclear Overhauser effect experiments showing the steric role of the 5 methylcytosine in the induction of an alternating B form in d(m5CpG)2. PMID- 3024978 TI - The transmembrane arrangement of the ADP/ATP carrier as elucidated by the lysine reagent pyridoxal 5-phosphate. AB - The lysine reagent pyridoxal 5-phosphate was applied to the ADP/ATP carrier (AAC) in order to elucidate topological and functional properties of the numerous lysines within the primary structure. To establish appropriate labeling conditions, the influence of pyridoxal-P on transport and inhibitor binding to the AAC was examined. The ADP/ATP transport is sensitive to low concentrations of pyridoxal-P with a Ki = 0.4 mM. Binding of [3H]carboxyatracylate and [3H]bongkrekate is largely inhibited by pyridoxal-P treatment with Ki approximately 1 mM. [3H]Carboxyatractylate is not and [3H]bongkrekate weakly removed by pyridoxal-P, whereas [3H]atractylate is displaced to a large extent. Under optimized conditions of pyridoxal-P concentration, of pH and of time exposure, the AAC was exposed to [3H]pyridoxal-P in mitochondria, in submitochondrial particles and in the detergent-solubilized carrier. The [3H]pyridoxal-P-labeled AAC was isolated from mitochondria and particles. After citraconylation thermolysinolytic peptides were prepared. The pyridoxyl-lysine containing peptides were purified and the pyridoxal-P incorporation to specific lysines was determined by sequencing. The pyridoxal-P incorporation into the AAC in various states was evaluated with regard to structural and functional aspects. First, by comparing pyridoxal-P incorporation in mitochondria and sonic particles, the segments of the polypeptide chain exposed to the cytosolic and matrix side of the membrane are detected. Second, the additional lysine incorporation into the isolated as compared to the membrane-bound carrier is attributed to the protein collar facing the phospholipid headgroups. Third, the difference between lysine incorporation into the carboxyatractylate-AAC and bongkrekate-AAC complexes reflect either conformational changes or lysines involved in the translocation channel through the protein. Fourth, the additional lysine labeled in the atractylate-carrier complex as compared to the carboxyatractylate-carrier complex is attributed to a cationic site in the binding center. These results are incorporated into a transmembrane folding model of the carrier. PMID- 3024979 TI - Characterization of the elongation factors from calf brain. 3. Properties of the GTPase activity of EF-1 alpha and mode of action of kirromycin. AB - The GTPase activity of purified EF-1 alpha from calf brain has been studied under various experimental conditions and compared with that of EF-Tu. EF-1 alpha displays a much higher GTPase turnover than EF-Tu in the absence of aminoacyl tRNA (aa-tRNA) and ribosomes (intrinsic GTPase activity); this is due to the higher exchange rate between bound GDP and free GTP. Also the intrinsic GTPase of EF-1 alpha is enhanced by increasing the concentration of monovalent cations, K+ being more effective than NH+4. Differently from EF-Tu, aa-tRNA is much more active than ribosomes in stimulating the EF-1 alpha GTPase activity. However, ribosomes strongly reinforce the aa-tRNA effect. In the absence of aa-tRNA the rate-limiting step of the GTPase turnover appears to be the hydrolysis of GTP, whereas in its presence the GDP/GTP exchange reaction becomes rate-limiting, since addition of EF-1 beta enhances turnover GTPase activity. Kirromycin moderately inhibits the intrinsic GTPase of EF-1 alpha; this effect turns into stimulation when aa-tRNA is present. Addition of ribosomes abolishes any kirromycin effect. The inability of kirromycin to affect the EF-1 alpha/guanine nucleotide interaction in the presence of ribosomes shows that, differently from EF-Tu, the EF-1 alpha X GDP/GTP exchange reaction takes place on the ribosome. PMID- 3024981 TI - Analysis of the puromycin reaction. The ribosomal exclusion principle for AcPhe tRNA binding re-examined. AB - The standard technique for determination of the ribosomal site location of bound tRNA, viz. the puromycin reaction, has been analyzed with regard to its applicability under tRNA saturation conditions. The criteria derived have been used to re-examine the exclusion principle for peptidyl-tRNA binding, which states that only one peptidyl-tRNA (AcPhe-tRNA) can be bound per ribosome although in principle two sites (A and P site) are available. The following results were obtained. The puromycin reaction is only appropriate for a site determination if the reaction conditions prevent one ribosome from performing more than one puromycin reaction. With an excess of AcPhe-tRNA over ribosomes, and in the absence of EF-G, this criterion is fulfilled at 0 degree C, where the P-site-bound material reacts with puromycin (quantitative reaction after 50 h), while the A-site-bound material does not. In contrast, at 37 degrees C the extent of the puromycin reaction can exceed the binding values by 2-4-fold ('repetitive reaction'). In the presence of EF-G a repetitive puromycin reaction is seen even at 0 degree C, i.e. EF-G can already promote a translocation reaction at 0 degree C. However, the extent of translocation becomes negligibly low for short incubation times (up to 60 min) at 0 degree C, if only catalytic amounts of EF-G are used. Using the criteria outlined above, the validity of the exclusion principle for Escherichia coli ribosomes was confirmed pursuing two different experimental strategies. Ribosomes were saturated with AcPhe-tRNA at one molecule per 70S ribosome, and a quantitative puromycin reaction demonstrated the exclusive P-site location of the AcPhe-tRNA. The same result was also found in the presence of viomycin, which blocks the translocation reaction. These findings also indicate that here nearly 100% of the ribosomes participate in AcPhe-tRNA binding to the P site. Precharging the P sites of 70S ribosomes with one Ac[14C]Phe-tRNA molecule per ribosome prevented additional Ac[3H]Phe-tRNA binding. In contrast, 70S particles carrying one molecule of [14C]tRNAPhe per ribosome were able to bind up to a further 0.64 molecule Ac[3H]Phe-tRNA per ribosome. PMID- 3024980 TI - Chemical modification of the brown-fat-mitochondrial uncoupling protein with tetranitromethane and N-ethylmaleimide. A cysteine residue is implicated in the nucleotide regulation of anion permeability. AB - Treatment of brown adipose tissue mitochondria with tetranitromethane or N ethylmaleimide decreases the affinity with which inhibitory nucleotide GDP binds to the tissue-specific uncoupling protein. Both reagents modify cysteine residues which are 'accessible' and 'buried' to 5,5'-dithio-bis(2-nitrobenzoic acid) (Nbs2). Modification of the single Nbs2-accessible residue correlates with the loss of high-affinity binding sites for GDP. Tetranitromethane does not affect the Cl- or H+ permeability of the protein in the absence of nucleotide, while N ethylmaleimide increases both by 70-80%. Bound GDP is a less effective inhibitor of Cl- permeability after N-ethylmaleimide or tetranitromethane treatment, but retains much of the ability to inhibit H+ permeation. PMID- 3024982 TI - Cooperative DNA binding by lambda integration protein--a key component of specificity. AB - Quantitative analysis of nitrocellulose filter binding data by the method of Clore, Gronenborn and Davies [(1982) J. Mol. Biol. 155, 447-466] has been used to show that lambda integration protein (Int) exhibits cooperativity in binding to specific recognition sites within the attachment site region (lambda attP) of bacteriophage lambda DNA. Optimal values of the equilibrium constant obtained were 3.0(+/- 1.0) X 10(10) M-1 for the P' site using a model of three sites with equal affinity and 1.9(+/- 0.4) X 10(10) M-1 for the P1 site on a two-site model. The value of the cooperativity parameter alpha is 172(+106)(-66) in all cases. The occurrence of a consensus recognition sequence is necessary but not sufficient for strong binding; cooperative interaction between Int molecules binding to adjacent members of an array of binding sites is also essential. The occurrence of binding site arrays distinguishes lambda attP very clearly from other DNA sequences containing single recognition sites by chance. PMID- 3024983 TI - Fructose 2,6-bisphosphate in Dictyostelium discoideum. Independence of cyclic AMP production and inhibition of fructose-1,6-bisphosphatase. AB - The occurrence of fructose 2,6-bisphosphate was detected in Dictyostelium discoideum. The levels of this compound were compared with those of cyclic AMP and several glycolytic intermediates during the early stages of development. Removal of the growth medium and resuspension of the organism in the differentiation medium decreased the content of fructose 2,6-bisphosphate to about 20% within 1 h, remaining low when starvation-induced development was followed for 8 h. The content of cyclic AMP exhibited a transient increase that did not correlate with the change in fructose 2,6-bisphosphate. If after 1 h of development 2% glucose was added to the differentiation medium, fructose 2,6 bisphosphate rapidly rose to similar levels to those found in the vegetative state, while the increase in cyclic AMP was prevented. The contents of hexose 6 phosphates, fructose 1,6-bisphosphate and triose phosphates changed in a way that was parallel to that of fructose 2,6-bisphosphate, and addition of sugar resulted in a large increase in the levels of these metabolites. The content of fructose 2,6-bisphosphate was not significantly modified by the addition of the 8-bromo or dibutyryl derivatives of cyclic AMP to the differentiation medium. These results provide evidence that the changes in fructose 2,6-bisphosphate levels in D. discoideum development are not related to a cyclic-AMP-dependent mechanism but to the availability of substrate. Fructose 2,6-bisphosphate was found to inhibit fructose-1,6-bisphosphatase activity of this organism at nanomolar concentrations, while it does not affect the activity of phosphofructokinase in the micromolar range. The possible physiological implications of these phenomena are discussed. PMID- 3024984 TI - Phosphorylation of the glycogen-binding subunit of protein phosphatase-1G by cyclic-AMP-dependent protein kinase promotes translocation of the phosphatase from glycogen to cytosol in rabbit skeletal muscle. AB - The glycogen-bound form of protein phosphatase-1 (termed protein phosphatase-1G) is composed of the catalytic (C) subunit complexed to a glycogen-binding (G) subunit that anchors the enzyme to glycogen [Stralfors et al. (1985) Eur. J. Biochem. 149, 295-303]. Incubation of purified protein phosphatase-1G with cyclic AMP-dependent protein kinase and MgATP, which leads to stoichiometric phosphorylation of the G-subunit [Caudwell et al. (1986) FEBS Lett. 194, 85-90], was found to promote the release of the phosphatase from glycogen; similar observations were made using glycogen-protein particle preparations. An intravenous injection of adrenaline decreased protein phosphatase-1 activity associated with the glycogen-protein particles by 50% with a corresponding increase in the amount present in the cytosol. By contrast, adrenaline did not affect the distribution of glycogen synthase or glycogen phosphorylase which remained entirely bound to glycogen in these experiments. The specific release of protein phosphatase-1 from glycogen may facilitate its inactivation by inhibitor 1 in the cytosol, thereby preventing dephosphorylation of the glycogen metabolising enzymes. Translocation of protein phosphatase-1 may represent a novel mechanism for the activation of glycogenolysis and inhibition of glycogen synthesis by adrenaline. PMID- 3024985 TI - Assessment of SPECT ventilation-perfusion imaging in patients with lung cancer. AB - Ventilation and perfusion SPECT images during tidal breathing were studied in 15 cases of lung cancer using 81mKr gas and 99mTc-microspheres. Furthermore, functional images of V/Q ratio and Q/V ratio were prepared, and their clinical significance is discussed with reference to general lung function. There was a decrease in %VC and %FEV 1.0 in 7 of 15 cases, and an increase of AaDo2 in the blood gas analysis in 12 of 15 cases. Both planar and SPECT images showed ventilation and perfusion abnormalities in all 15 cases. Of these, 12 patients showed matched ventilation and perfusion defects, 2 patients a dead-space effect and 1 patient a shunt effect. In comparing planar and SPECT images, depiction of ventilation and perfusion impairments were equally clear in 11 cases, but in 3, showing a lobar or segmental defect with a shunt effect, the SPECT images were superior. In a patient with markedly impaired function of the affected lung, the remaining function could not be depicted by SPECT. From the above, it seems that better information can be obtained for understanding the ventilation and perfusion states of lung cancer by adding the SPECT images to the planar image. PMID- 3024986 TI - 99mTc-methylene diphosphonate uptake by ossifications and calcifications of non osseous metastatic tumors. AB - Two cases of extra-osseous uptake of 99mTc-methylene diphosphonate (MDP) by non osseous metastatic tumors are reported. One was a metastasis with ossification in the abdominal wall from carcinoma of the sigmoid colon and the other was a metastasis with calcification from an ovarian carcinoma. The mechanism of extra osseous uptake of 99mTc-MDP is discussed. Bone scintigraphy can be a potential means to assess tumor spread with ossifications and calcifications. PMID- 3024988 TI - Systemic lupus erythematosus presenting with recurrent acute demyelinating polyneuropathy. AB - We report a patient who in the course of 3 years developed 3 distinct episodes of an acute demyelinating neuropathy, each fulfilling the criteria set up by an ad hoc committee of the National Institute of Neurological and Communicative Disorders and Stroke for acute Guillain-Barre syndrome. Each episode was associated with strong serological evidence for lupus erythematosus. The episodes were separated by 1-2 years and each was followed by near complete clinical recovery. The only systemic feature was a pleural effusion during the last 2 episodes. PMID- 3024987 TI - Vitamin D-dependent rickets type II: extreme end organ resistance to 1,25 dihydroxy vitamin D3 in a patient without alopecia. AB - Vitamin D-dependent rickets type II (VDDR II) is a rare syndrome resulting in severe rickets and is resistant to treatment with vitamin D and its derivatives. Patient with this disease, who are frequently the children of consanguinous marriages, present with elevated circulating concentrations of 1,25-dihydroxy vitamin D, the active metabolite of vitamin D, and in vitro studies have indicated a failure of intracellular binding of the hormone. Alopecia has been noted in many of these patients and it has been suggested that this feature may indicate a more marked resistance to treatment. However we describe a 3-year-old boy with this disease who, although having normal hair growth, displayed extreme resistance to treatment with active vitamin D metabolites. In vitro studies of skin fibroblasts disclosed not only an absence of hormone binding or 1,25(OH)2D3 induced 24-hydroxylase activity but reduced metabolism of 1,25(OH)2D3 itself. In this child, treatment with exogenous 1,25-dihydroxy vitamin D3 at doses of up to 24 micrograms/day, which increased the circulating concentration of the metabolite to greater than 100 times the normal adult mean, failed to alleviate his condition and he died at the age of 39 months. This would therefore suggest that absence of alopecia, in this condition, cannot be regarded as a constant predictive sign of a lesser resistance and of responsiveness to Vitamin D treatment. PMID- 3024989 TI - Motor neuropathy with proximal multifocal persistent conduction block, fasciculations and myokymia. Evolution to tetraplegia. AB - We describe a patient with chronic asymmetric motor neuropathy, which began in the upper extremity. The paretic muscles showed abundant fasciculations and myokymia but only little amyotrophy. Electrophysiologic examination revealed proximal multifocal persistent conduction block (CB) not located at the usual entrapment sites, and arrhythmic isolated or grouped fasciculation potentials originating distally on blocked axons. Over the years, new CBs developed, which led to tetraplegia, and amyotrophy slowly increased with progressive denervation. This patient differs from the cases of chronic acquired demyelinating polyneuropathy described in the literature by the absence of sensory deficit and the proximal location of CB. PMID- 3024990 TI - Styrene-induced peripheral neuropathy. A case report. AB - A case of peripheral neuropathy due to styrene is described. The diagnosis of peripheral neuropathy was confirmed by a motor-nerve conduction study and sural nerve biopsy. PMID- 3024991 TI - Emetic and antiemetic effects of opioids in the dog. AB - The emetic and antiemetic effects of opioid agonists were studied in awake dogs. The mu-agonists morphine, fentanyl and methadone, in sedative doses, prevented the emetic response to apomorphine and copper sulphate; only morphine induced emesis, at doses lower than those required to prevent emesis. The delta-agonist [D-Ala2,Met5]enkephalinamide (DALA) and [Leu5]enkephalin induced emesis in some of the dogs studied but had no antiemetic activity. The kappa-agonists bremazocine and ethylketocyclazocine (EKC) did not induce emesis but, at sedative doses, prevented the emetic response to apomorphine. The emetic effect of DALA was antagonized by naloxone in some dogs; the antiemetic effect of morphine, bremazocine and EKC was blocked by both naloxone and MR 2266. The non-opioid sedatives diazepam, phenobarbital and xylazine, administered in sedative doses, did not prevent apomorphine-induced emesis. Our results suggest that a delta receptor is involved in the emetic effect and a mu- and/or or kappa-receptor in the antiemetic effect of opioids. PMID- 3024992 TI - Multiple tachykinin binding sites in hamster, rat and guinea-pig urinary bladder. AB - Binding of the 125I-Bolton-Hunter labelled tachykinins substance P, substance K, eledoisin and neuromedin K (BHSP, BHSK, BHE, BHNK) was examined in urinary bladders of hamster, rat and guinea-pig using crude membrane suspensions and by autoradiography. High-affinity binding of BHSK was observed in hamster and rat bladder and high-affinity binding of BHSP was seen in rat and guinea-pig bladder. Characterization of this binding showed that the hamster bladder contains very large numbers of K-type binding sites, where BHSK is displaced by substance K greater than kassinin greater than eledoisin greater than neuromedin K greater than substance P greater than physalaemin, and has very few P-type binding sites, where BHSP is displaced by substance P greater than substance K much greater than neuromedin K. In contrast, the rat bladder contains moderate and approximately equal numbers of both K (KD, 0.74 nM; Bmax 2.9 fmol/mg wet weight tissue) and P (KD, 0.12 +/- 0.01 nM; Bmax 2.6 +/- 0.2 fmol/mg wet weight tissue) sites. The guinea-pig bladder possesses predominantly P sites. Most tachykinin binding sites are localized over smooth muscle and probably represent functional receptors mediating the direct contractile effects of tachykinins in these tissues. Few E type binding sites, as previously described in rat brain, were found, although some BHNK binding sites were seen in the mucosa of guinea-pig bladder. PMID- 3024993 TI - Different types of relationship between beta-adrenergic relaxation and activation of cyclic AMP-dependent protein kinase in canine saphenous and portal veins. AB - We examined the relationships between the relaxation mediated by beta adrenoceptor and either the associated cyclic AMP production or the activation of cyclic AMP-dependent protein kinase (A-PK) in canine saphenous and portal veins. In the saphenous vein constricted with methoxamine, isoproterenol caused concentration-dependent relaxation (maximum relaxation 92.7%), and concomitant increases in cyclic AMP and A-PK activity ratio (from 52.8 to 73.5%). The portal vein was only slightly relaxed by isoproterenol (14.7% in the longitudinal strips) after constriction with methoxamine, while cyclic AMP and A-PK activation increased significantly. Isoproterenol markedly activated A-PK of the portal vein after KCl constriction (from 52.6 to 74.6%) but the maximum relaxation was only slight (13.3%). The portal vein also showed a smaller relaxation response to either forskolin or dibutyryl cyclic AMP than the saphenous vein. These results indicated that the relationship between the relaxation response to isoproterenol and either cyclic AMP production or A-PK activation was different in the saphenous and portal veins. PMID- 3024994 TI - Evaluation of 4-methylpiperidine analogs of hemicholinium-3. AB - A series of substituted piperidine analogs of hemicholinium-3 was evaluated for their ability to inhibit neuromuscular transmission, to decrease acetylcholine content of caudate slices, to inhibit choline acetyltransferase activity, and to produce toxicity. Quaternary and tertiary amine derivatives of 4-methyl- and 4 hydroxyl-substituted piperidine analogs containing beta-carbonyl or beta-hydroxyl substitutions in the phenylethyl spacing moiety were tested. 4-Methyl piperidine derivatives maintained potent hemicholinium-3 like activity. Reduction of activity was seen with the 4-hydroxyl piperidine analogs. Compounds with beta hydroxyl substitution were more potent than those with beta-carbonyl substitution. The tertiary amine, 4-methyl piperidine derivative with a hydroxyl group on the beta-carbon of the ethyl side chain also possessed hemicholinium-3 like activity. However, tertiary amine analogs were substantially less potent than hemicholinium-3 or their quaternary amine analogs. PMID- 3024995 TI - Pharmacological evidence for the existence of interactions between dopaminergic and opioid peptidergic systems in guinea-pig ileum myenteric plexus. AB - Apomorphine or bromocriptine treatment at doses that act on dopamine autoreceptors resulted in a significant elevation of the release of opioid peptides from the myenteric plexus of guinea-pig, since these drugs produced an increase of the inhibitory response which was reversed by naloxone. 6 Hydroxydopamine treatment also resulted in an increase in opioid peptide release. These findings would indicate that the interruption of dopaminergic transmission in the myenteric plexus produces an increase in the release of opioid peptides and suggest an inhibitory modulation of opioid peptidergic neurons by dopamine systems at this level. PMID- 3024996 TI - Stereoselective inhibition of the binding of [3H]PK 11195 to peripheral-type benzodiazepine binding sites by a quinolinepropanamide derivative. AB - The specific binding of [3H]PK 11195 to the peripheral-type benzodiazepine binding site is inhibited by the l-enantiomer of N,N-diethyl-alpha-methyl-2 phenyl-4-quinolinepropanamide ((-)Q1) but not by its d-enantiomer ((+)Q1). (-)Q1 inhibited [3H]PK 11195 binding to several rat tissues with an IC50 of less than 10 nM whereas (+)Q1 was at least 500 times less potent. This stereoselectivity was observed in all the tissues tested (brain, heart, kidney and adrenals). The same stereoselectivity was found for the displacement of the binding of [3H]PK 11195 in vivo, where (-)Q1 had an ID50 between 4-15 mg/kg and (+)Q1 was completely inactive at all doses tested (i.e. up to 40 mg/kg). Neither isomer had appreciable affinity for central-type benzodiazepine binding sites ([3H]diazepam) nor for voltage-sensitive calcium channels ([3H]PN 200210 and [3H]verapamil). PMID- 3024997 TI - Inhibitory action of capsaicin on cholinergic responses induced by GABAA agonists in the guinea-pig ileum. AB - Pretreatment of the guinea-pig ileum with capsaicin resulted consistently in depression of the neurogenic cholinergic contractions induced by the GABAA receptor agonists 3-aminopropane sulphonic acid (3-APS) and muscimol. Since capsaicin acts mainly by releasing and depleting substance P from its stores in intestinal nerves, it is likely that substance P plays a role in the response caused by GABAA-mimetic compounds, On the whole, our results suggest that excitatory responses to 3-APS and muscimol result from both direct and indirect activation of intrinsic intestinal cholinergic neurons innervating smooth muscle cells. PMID- 3024998 TI - Ontogenesis of delta-opioid receptors in rat brain using [3H][D-Pen2,D Pen5]enkephalin as a binding ligand. AB - The ontogenesis of delta-opioid receptors has been studied in the postnatal period up to day 50 using the highly selective delta-site ligand [3H][D-Pen2,D Pen5]enkephalin ([3H]DPDPE) in binding studies. Analyses of saturation curves revealed marked increases in binding capacities between the second and fourth postnatal week with little change in affinity. In contrast to findings with [3H][D-Ala2,D-Leu5]enkephalin binding could not be detected before postnatal day 10 which may be associated with the low affinity and specific activity of DPDPE. A low specific binding for [3H]DPDPE poses methodological problems in the use of this ligand for ontogenic studies. PMID- 3025000 TI - [D-Pen2,D-Pen5]enkephalin (DPDPE): a delta-selective enkephalin with low affinity for mu 1 opiate binding sites. PMID- 3024999 TI - 8-Bromo-cyclic GMP abolishes TEA-induced slow action potentials in canine trachealis muscle. AB - Using intracellular microelectrodes, we investigated whether 8-bromo-guanosine 3':5'-cyclic monophosphate (cGMP) influenced the electromechanical effects of tetraethylammonium (TEA) on canine tracheal smooth muscle. We found that 20 mM TEA depolarized airway smooth muscle cells from -58 +/- 3 mV (means +/- S.D.) to 44 +/- 2 mV and caused spontaneous action potentials (APs) to develop which were 31 +/- 2 mV in amplitude. These APs, and the phasic contractions electrically coupled to them, were totally abolished in buffer containing 0.1 mM cGMP. Our findings suggest that cGMP markedly affects the channels mediating TEA-induced APs in airway smooth muscle. PMID- 3025001 TI - Effects of memantine on the frog neuromuscular junction. AB - The effects of memantine, an adamantane derivative, on neuromuscular transmission in the frog sartorius muscle preparation were studied by measuring the endplate current (EPC) by the voltage clamp method. Memantine (0.5-50 microM) reduced the peak amplitude and shortened the duration of the EPC, and the membrane voltage peak EPC relationship became non-linear. Since it decreased the quantal height of the EPC without affecting the mean quantal content, the effects were considered to be mainly postsynaptic. Noise analysis revealed that memantine decreased the mean lifetime of the ACh-activated channel. In addition, lineweaver-Burk analysis of the response to ionophoretically applied carbamylcholine showed that memantine acted essentially as a linear mixed-type inhibitor. Thus memantine seems to block neuromuscular transmission by reacting with the ACh receptor-ion channel complex in both the open and closed states. PMID- 3025002 TI - Opioid receptor types and antinociceptive activity in chronic inflammation: both kappa- and mu-opiate agonistic effects are enhanced in arthritic rats. AB - The antinociceptive effects obtained in arthritic rats with morphine, the opioid mu-agonist DAGO [D-Ala2,MePhe4,Gly-ol5]enkephalin, the delta-selective agonist DTLET [D-Thr2, Leu5]enkephalyl-Thr, and the kappa-agonist U-50,488H were compared to their corresponding effects in normal animals and morphine-pretreated arthritic rats, respectively, using a paw pressure test. The effects of the mu- and kappa-agonists were increased in arthritic rats. While morphine-treated rats were cross-tolerant to the mu- and kappa-agonists, no tolerance to the delta selective agonist was found. The possibility that the potent action of morphine in this model for chronic inflammatory pain is mediated partly through kappa mechanisms is discussed. PMID- 3025003 TI - The action of trelibet, a new antidepressive agent on [3H]noradrenaline release from rabbit pulmonary artery. AB - Trelibet, a new antidepressant, used at 10(-7)-10(-4) M failed to affect the [3H]noradrenaline ([3H]NA) release evoked from the isolated main pulmonary artery of the rabbit low frequency (2 Hz) nerve stimulation whether the neuronal uptake inhibitor cocaine (3 X 10(-5) M) was present or not. Its metabolite (EGYT-2760) however, potentiated the nerve-evoked release of [3H]NA. In the absence of cocaine both the resting and the stimulation-evoked release of 3H increased in response to EGYT-2760. These effect were accompanied by muscle contraction. The EGYT-2760-potentiated transmitter release was inhibited either by exogenously applied 1-noradrenaline (10(-6) M) or clonidine (10(-6) M), preferential agonists of presynaptic alpha 2-adrenoceptors. The 1-noradrenaline-induced inhibition of transmitter release potentiated by EGYT-2760 was antagonized by 3 X 10(-7) M yohimbine, a preferential alpha 2-adrenoceptor inhibitor. In the absence of cocaine, Ca2+ removal from the external medium failed to affect the 3H outflow increasing effect of EGYT-2760 but abolished the nerve-evoked release potentiating action of this compound. It is concluded that the metabolite of trelibet exerts a 'yohimbine-like' action, as well as a 'tyramine-like' effect in peripheral sympathetic nerve fibres. PMID- 3025004 TI - Ca-channel blockers and the electrophysiology of synaptic transmission of the guinea-pig olfactory cortex. AB - Slices of guinea-pig olfactory cortex have been used to compare the potency of various Ca-blockers on the electrophysiology of synaptic transmission. Listed in the order of potency, the divalent cations Cd2+, Ni2+, Mn2+, Co2+, La3+ and Mg2+ depressed synaptic transmission. The organic Ca-blockers, nifedipine or nimodipine or verapamil and diltiazem were ineffective up to 0.01 mmol/l. Verapamil, D600 or diltiazem (0.1-0.3 mmol/l) depressed both synaptic transmission and the sodium-mediated presynaptic action potential. These results reaffirm the idea that 'organic Ca-antagonist' do not block all Ca-channels in brain and the high Cd2+ sensitivity suggests the Ca-channels in post- and presynaptic membranes have dissimilar pharmacological profiles. PMID- 3025006 TI - Reduction of striatal dopaminergic neurotransmission elevates striatal proenkephalin mRNA. PMID- 3025005 TI - Photoaffinity labeling of neurotensin binding sites on rat brain sections. AB - The photoaffinity labeling of neurotensin (NT) binding sites was carried out on rat midbrain sections using a monoiodo analogue of NT (125I-azidobenzoyl [Trp11] NT; 125IAB-NT). Autoradiographic data showed that the 125IAB-NT binding site localization was quite similar to that obtained with 125I-NT, with high densities in both substantia nigra and ventral tegmental area. Covalent specific binding was only observed when sections were irradiated with UV after the incubation, followed by various histological treatments necessary for light and electron microscopy. PMID- 3025007 TI - Rat lateral septum in slice preparation with viable transmission. AB - A procedure is described which makes it possible to prepare slices from the rat brain that comprise nearly the entire lateral septum together with the major part of the fimbria-fornix afferent fibers to the lateral septum. The field potentials and monosynaptic excitation of lateral septal neurons elicited by electrical stimulation of the fimbria-fornix afferent fibers were employed to demonstrate that the neurophysiological features of the lateral septal neurons in vitro are similar to those found previously in vivo, and the synaptic transmission between fimbria-fornix afferent fibers and neurons of the lateral septum could be maintained in vitro without significant alterations for at least a 6-h period of incubation. PMID- 3025008 TI - Facilitatory effects of dexamethasone on neuromuscular transmission. AB - The beneficial effects of glucocorticoids in myasthenia gravis are attributed to their immunosuppressive actions. There are also studies reporting direct facilitatory as well as depressant effects of glucocorticoids on neuromuscular transmission. The effects of dexamethasone on neuromuscular transmission were studied by intracellular and extracellular microelectrode recording techniques in the mouse phrenic nerve-diaphragm preparation. Creatinine had to be added to the bathing media to prevent precipitation of the glucocorticoid with Ca2+ and Mg2+; creatinine had no effect. One hour of perfusion with dexamethasone (10(-4) to 10( 3) M) increased the frequency of miniature end-plate potentials (MEPPs), as well as the amplitude and quantum content of end-plate potentials (EPPs), but did not change MEPP amplitude, suggesting an increase in acetylcholine release. Dexamethasone also enhanced presynaptic facilitation and potentiation during repetitive stimulation. It had no effect on muscle resting membrane potential but increased the amplitude, overshoot, and rate of rise of muscle action potentials. The amplitudes of nerve terminal action potentials were also enhanced by dexamethasone. These findings suggest that glucocorticoids have a direct facilitatory action on neuromuscular transmission by a presynaptic action. PMID- 3025009 TI - Myocyte cytochrome oxidase activity and neuromuscular conduction associated with metabolic alkalemia. AB - Many studies have shown increased affinity of hemoglobin for oxygen during metabolic alkalemia and dependence of intramitochondrial cytochrome oxidase activity on arterial oxyhemoglobin saturation. The present studies tested the hypothesis that metabolic alkalemia produces tissue hypoxia independent of arterial oxygen desaturation. Neuromuscular conduction latency was used as an indicator of functional impairment, and was measured following electrostimulation of the sciatic nerve and recording of the electromyogram from the gastrocnemius muscle of rats anesthetized with pentobarbital sodium. To increase the affinity of hemoglobin for oxygen, sodium bicarbonate was administered in graded doses every 15 min. Statistical significance of changes was estimated by the paired Student's t test. Arterial bicarbonate ion concentration increased from 25 +/- 1.3 to 39.0 +/- 3.0 mM while arterial pH increased from 7.30 +/- 0.02 to 7.50 +/- 0.03 (P less than 0.01). Neuromuscular conduction latency increased from 1.9 +/- 0.13 to 2.7 +/- 0.18 ms (P less than 0.01). Tissue hypoxia was suggested by a greater decrease in mass spectrometric determinations of gastrocnemius muscle oxygen tension (PO2) in separate groups of control (arterial pH 7.38 +/- 0.04) and experimental (arterial pH 7.48 +/- 0.03) rats. These changes were accompanied by markedly decreased uptake of 3.3'-diaminobenzidine (DAB) by gastrocnemius muscle mitochondria, suggesting decreased intracellular activity of cytochrome oxidase and intracellular oxygen availability to myocytes in the absence of arterial oxygen desaturation. PMID- 3025010 TI - Alterations in beta-adrenergic receptor binding in partially and fully amygdala kindled juvenile and adult rats. AB - Twenty-eight-day-old (juvenile) rats were kindled with hourly stimulations to partial or fully kindled status. Adult rats were stimulated with hourly or daily stimulations. Alterations in [3H]dihydroalprenolol binding were determined 3 weeks after the last stimulation. We found that partially kindled, hourly stimulated juvenile rats showed a significant increase in the dissociation constant (Kd), with no change in maximal binding values. Fully kindled juvenile rats showed no change in Kd or Bmax. Partially kindled, hourly stimulated adult rats showed a significant decrease in Kd, with no change in Bmax. There was no change in Kd or Bmax values in fully kindled, hourly stimulated adult rats. Fully kindled, daily stimulated adult rats showed a decrease in maximal binding, with no change in Kd values. These findings indicate that kindling-induced beta adrenergic receptor alterations were influenced by the age of the animal and the kindling parameters used, as well as the extent to which the animals were kindled. PMID- 3025011 TI - Relative activities of 5'-nucleotidase and adenosine deaminase in atrial and ventricular myocardium--the enzyme paradox. AB - The 5'-nucleotidase and adenosine deaminase activities were determined for tissue extracts of up to 10 defined regions of the normal dog heart. The specific activity of 5'-nucleotidase was very significantly higher in atrial than in ventricular myocardium (4 X) and was also higher in right than in left sites of the heart (2 X). The reason for the markedly higher specific activity in atrial muscle is not clear and appears paradoxical relative to adenine nucleotide content and pattern of ATP decay during oxygen deprivation. In contrast to 5' nucleotidase activity the total adenosine deaminase activity was found to be similar in all sampling sites examined. PMID- 3025012 TI - Leukocyte mediated vein injury and thrombosis is reduced by a lipoxygenase inhibitor. AB - We have previously demonstrated an in vivo model of deep vein thrombosis which suggests that the neutrophil promotes vascular injury and thrombosis following blood flow stasis. Since leukotrienes are potent mediators of vascular injury and neutrophil (PMN) chemotaxis, we wished to determine if in vivo inhibition of 5 lipoxygenase would reduce neutrophil mediated events in our model. Lipoxygenase was inhibited in vivo with 2,3-diethyl-4-methoxy,1-naphthalenol acetate (U 66,855). The in vivo activity of U-66,855 was demonstrated in 4 cats. Each animal was treated with 5 mg/kg of U-66,855 intravenously. Blood cell leukotriene B4 (LTB4) and thromboxane A2, via its metabolite thromboxane B2 (TBX2) was assessed before and 30, 60, and 120 min after dosing. Blood cell LTB4 and TBX2 production was stimulated by A23187 (24 microM) and assayed by radioimmunoassay. We exposed and isolated a 3-cm segment of the jugular veins from 10 additional cats 5 of which were treated with U-66,855 (5 mg/kg, iv). In order to assess the effect of stasis, the jugular veins were ligated at the thoracic inlet for 2 hr after which the veins were perfused, fixed in 2.5% glutaraldehyde, and prepared for electron microscopy. U-66,855 reduced LTB4 production significantly (P less than 0.01), but not TBX2. In untreated cats, PMNs adhered to and migrated underneath the venous endothelium. Additionally, platelets, fibrin and formed thrombi were found on the basement membrane exposed by the migrating neutrophils. In contrast, we observed significantly reduced PMN adhesion as well as no fibrin deposition in veins obtained from cats treated with U-66,855. The results suggest that 5 lipoxygenase inhibition can significantly reduce undesirable neutrophil/vessel wall interactions. PMID- 3025013 TI - Poliomyelitis immunity status at different intervals from vaccination. AB - Research on neutralizing antibodies against the 3 serotypes of poliovirus was carried out on 441 blood specimens drawn from children and young people living in Parma, a town of about 200,000 inhabitants, where the vaccination services were well organized. From June 1983 to June 1984 blood samples were taken from different groups of subjects at different stages of vaccination (before vaccination, and after 1, 2, 3, and 4 doses of trivalent balanced OPV). In addition, groups of children and young people vaccinated from 2 to more than 14 years earlier were bled. Ten per cent of unvaccinated babies (aged 1-3 months) were lacking in neutralizing antibodies for types 1 and 3 and five per cent for type 2; among children aged 4-5 months vaccinated by one dose of OPV a residual 5% of subjects without antibodies for type 1 was also found. All subjects who received two, three, or four doses, had antibodies against all serotypes of poliovirus. As for the number of administered doses, significant differences among GMTs of antibodies were recorded only for type 1, when the group given 3 doses, was compared with the group which received two and one dose, respectively. At different intervals from completion of vaccination (2-14 years), all groups we examined still had antibodies against all three serotypes. For poliovirus type 1 antibody, GMTs were not significantly different in the examined groups. For types 2 and 3 significant decreases were observed in the groups vaccinated from 2 to 5 years. The high levels of immunity observed earlier in Parma were not found throughout the entire territory.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3025014 TI - Phosphorylation of the basal site of hormone-sensitive lipase by glycogen synthase kinase-4. AB - In rat adipocytes hormone-sensitive lipase is phosphorylated at two sites termed 'regulatory' and 'basal', in the former case by cyclic AMP-dependent protein kinase causing an activation of the lipase [(1984) Proc. Natl. Acad. Sci. USA 81, 3317-3321]. Here, the basal phosphorylation site was found to be phosphorylated by glycogen synthase kinase-4 without any effects on lipase activity, or on the extent of its activation subsequent to phosphorylation of the regulatory site. Glycogen synthase kinase-3, casein kinase-I, and casein kinase-II did not phosphorylate the lipase. Phosphorylase kinase phosphorylated it to a very low extent at a third phosphorylation site not phosphorylated in the fat cell. PMID- 3025015 TI - The second pX product p27 chi-III of HTLV-1 is required for gag gene expression. AB - The human T-cell leukemia virus type-1 (HTLV-1) contains a unique pX region, which encodes the gene products p40 chi, p27 chi-III and p21 chi-III. p40 chi is required for transcriptional trans-activation, whereas p27 chi-III and p21 chi III have no such function. Transfection of pX expression plasmids containing different combinations for the three gene products into cells integrated with HTLV-1 proviruses defective in pX expression revealed that both p40 chi and p27 chi-III are required for expression of the gag protein and accumulation of gag mRNA. These observations suggest that the pX product p40 chi activates transcription and p27 chi-III controls the level of gag mRNA by post transcriptional modulation. PMID- 3025016 TI - Plasminogen activator inhibitor type-1: reactive center and amino-terminal heterogeneity determined by protein and cDNA sequencing. AB - Both the urokinase-type and tissue-type plasminogen activator can convert their approximately 54 kDa type-1 inhibitor (PAI-1) to an inactive form with a lower apparent molecular mass. We have determined the amino-terminal amino acid sequences of human native and converted PAI-1, and isolated PAI-1 cDNA and determined the nucleotide sequence in regions corresponding to the amino-terminus and the cleavage site. The data show that the conversion of the inhibitor consists of cleavage of an Arg-Met bond 33 residues from the carboxy-terminus, thus localizing the reactive center of the inhibitor to that position. In addition, a heterogeneity was found at the amino-terminus, with a Ser-Ala-Val-His His form and a two-residue shorter form (Val-His-His-) occurring in approximately equal quantities. PMID- 3025017 TI - Polyethylene glycol-stimulated microsomal GTP hydrolysis. Relationship to GTP mediated Ca2+ release. AB - It has recently been observed that GTP mediates Ca2+ release from internal Ca2+ stores. In contrast to effects on permeabilized cells, GTP-dependent Ca2+ release in isolated microsomes requires the presence of polyethylene glycol (PEG). We have investigated the effects of PEG on microsomal GTPase activity and report that PEG stimulates a high-affinity (Km = 0.9 microM) GTPase. The effects of PEG reflect an increase in the Vmax of this activity; no effects on Km were observed. The concentration dependence for PEG-dependent stimulation of the high-affinity GTPase exactly mimicked that for GTP-dependent Ca2+ release. The stimulation of GTP hydrolysis by PEG was specific for the microsome fraction; only small effects were obtained with plasma membrane or cytosol fractions. As observed for GTP dependent Ca2+ release, the microsomal PEG-stimulated GTPase was competitively inhibited by the GTP analog GTP gamma S (Ki = 60 nM). It is proposed that the PEG stimulated GTPase may represent an intrinsic activity of the guanine nucleotide binding protein involved in the regulation of reticular Ca2+ fluxes. PMID- 3025018 TI - Preliminary X-ray studies of the tetra-heme cytochrome c3 and the octa-heme cytochrome c3 from Desulfovibrio gigas. AB - A tetra-heme and an octa-heme cytochrome c3 from the sulfate bacterium Desulfovibrio gigas have been crystallized. Diffraction quality crystals of the tetra-heme cytochrome are obtained from solution by the addition of polyethylene glycol at pH 6.5. The crystals are orthorhombic, space group P2(1)2(1)2 with unit cell parameters a = 42.27 A, b = 52.54 A and c = 52.83 A. The octa-heme cytochrome crystals develop from low ionic strength solutions of phosphate or Tris-Cl in the pH range 6.2-7.6. The crystals belong to the trigonal system, space group P3(1) or the enantiomorph P3(2), with unit cell parameters a = b = 57.4 A, c = 97.3 A, gamma = 120 degrees. Single crystal diffraction studies of the structures of these two low-potential cytochromes are in progress. PMID- 3025019 TI - Identification of p26Xb and p24Xb of human T-cell leukemia virus type II. AB - Human T-cell leukemia virus type II (HTLV-II) isolated from a T-cell variant of hairy cell leukemia contains gag, pol and env genes as well as a fourth gene termed X, which can code three major open reading frames Xa, Xb and Xc. Proteins with molecular masses of 26 kDa (p26Xb) and 24 kDa (p24Xb) encoded by the Xb open reading frame were identified with antisera directed against synthetic peptides corresponding to the N-terminal and C-terminal amino acid sequences deduced from the structure of the Xb open reading frame. More than half the Xb products were found to be located in the nuclear fraction of HTLV-II-infected cells. PMID- 3025020 TI - Absence of a unique relationship between active transport of lactose and protonmotive force in E. coli. AB - The relationship between active transport of lactose via the lactose permease and the protonmotive force has been determined in E. coli cells using either the respiratory chain inhibitor cyanide or protonophores to decrease the protonmotive force progressively. In contradiction with the prediction of the delocalized chemiosmotic theory, two different relationships were obtained depending on the method used. PMID- 3025021 TI - Receptor binding, cGMP stimulation and receptor desensitization by atrial natriuretic peptides in cultured A10 vascular smooth muscle cells. AB - Receptor characteristics for atrial natriuretic peptides (ANP) were demonstrated in the permanent tissue culture system of vascular smooth muscle cell (VSMC) line. 125I-ANP exhibited reversible and saturable binding to A10 rat VSMC, and the equilibrium dissociation constant, Kd was 157 pM and maximal binding capacity, Bmax amounted to 115 fmol/mg of protein. Binding of the 125I-ligand was highly specific for certain potently displacing ANP analogues with inhibition constants (Ki values) in the nano- or even subnanomolar concentration range. Pretreatment of VSMC with ANP yielded receptor desensitization to one half of the ANP receptor density, and supports the conclusion of receptor autoregulation by ANP in A10 VSMC. The receptor coupled intracellular cGMP system was stimulated, and thus demonstrates the application of A10 cells as a model for the study of ANP receptor interaction involved in vascular smooth muscle relaxation. PMID- 3025022 TI - Regulation of mitochondrial proteolysis. Selective degradation of inner membrane polypeptides. AB - The in vitro degradation of respiratory chain polypeptide components by a proteinase associated with the intermembrane space fraction was studied in rat liver mitochondria. Differences in susceptibility to proteolysis were detected by gel analysis after electrophoretic separation of the degraded polypeptides. A 55 kDa subunit is protected from proteolysis by the ATP molecule. PMID- 3025023 TI - The expression in E. coli of a polymeric gene coding for an esterase mimic catalyzing the hydrolysis of p-nitrophenyl esters. AB - We have prepared a hybrid protein consisting of seven esterase units, Glu-Ala-His Ala-Ser-Phe-Phe-Phe, fused to the N-terminal of galactokinase (E. coli). The structural gene for this bifunctional protein was obtained by cloning a polymer made up of three chemically synthesized oligonucleotides to which the galactokinase gene was fused in frame. The hybrid protein was purified to homogeneity with the aid of the galactokinase moiety and showed an Mr of 51,000 53,000. The preparation could catalyze the hydrolysis of p-nitrophenyl esters and, due to the inbuilt hydrophobic spacers, Phe-Phe-Phe, improved catalysis of more hydrophobic substrates was obtained. PMID- 3025024 TI - Degradation of nuclear proteins: studies on transplanted B82 cell karyoplast proteins. AB - Karyoplasts were prepared from B82 cells (thymidine kinase deficient mouse L cells) by cytochalasin B mediated enucleation. Morphological measurements show that the nucleus constitutes 89% of a karyoplast by volume. Homokaryons were obtained by Sendai virus mediated karyoplast-B82 cell fusion. Transplanted nuclei were not destroyed catastrophically but were maintained intracellularly for at least 140 h. Transplanted nuclear proteins were degraded with an average half life of 84 +/- 7 h by processes partially sensitive to inhibition by NH4Cl (50%) and leupeptin (30%). The data therefore suggest that some nuclear proteins are translocated to the cytoplasm for lysosomal degradation. PMID- 3025025 TI - Phosphorylation of liver gap junction protein by protein kinase C. AB - The 27 kDa protein, a major component of rat liver gap junctions, was shown to be phosphorylated in vitro by protein kinase C. The stoichiometry of the phosphorylation indicated that approx. 0.33 mol phosphate was incorporated per mol 27 kDa protein. Phosphorylation was entirely dependent on the presence of calcium and was virtually specific for serine residues. For comparison, the gap junction protein was also examined for its phosphorylation by cAMP-dependent protein kinase, the extent of phosphorylation being one-tenth that exerted by protein kinase C. PMID- 3025026 TI - Oxidation of reduced cytochrome c by hydrogen peroxide. Implications for superoxide assays. AB - Hydrogen peroxide, formed directly or as a product of superoxide dismutation, can oxidize ferrocytochrome c at rates comparable to those at which ferricytochrome c is reduced by superoxide. This reoxidation can significantly affect estimates of rates and amounts of superoxide production using absorbance changes for cytochrome c at 550 nm as the assay. The oxidation can be inhibited by catalase. PMID- 3025028 TI - [Modern methods of diagnosing malignant neoplasms of the retroperitoneal space in children]. PMID- 3025027 TI - Carbachol and sodium fluoride, but not TSH, stimulate the generation of inositol phosphates in the dog thyroid. AB - In dog thyroid slices prelabeled with myo-[2-3H]inositol, carbachol (10(-7)-10( 4) M) and NaF (10-20 mM) stimulated IP1, IP2 and IP3 generation. These effects did not require the presence of extracellular calcium. Atropine and PDBu inhibited the action of the cholinergic agonist. No effect of TSH (1-100 mU/ml) could be detected on PIP2 hydrolysis and IP production. These results suggest that IP3 could play a role in the metabolic actions of carbachol in the thyroid; a G-protein coupling the hormone-receptor binding to phospholipase C activation exists in the thyroid membrane; the well known TSH-induced increased PI turnover does not result in IP3 accumulation. PMID- 3025029 TI - Purification of nuclear cAMP-independent protein kinases from rat ventral prostate. AB - Two nuclear cAMP-independent protein kinases (designated PK-N1 and PK-N2) were purified from rat ventral-prostate and liver. The yield of enzyme units was 4-5% and 7-9% for each enzyme from the prostatic nuclei and liver nuclei, respectively. The average fold purification for prostatic nuclear protein kinase N1 and N2 was 1360 and 1833, respectively. The respective average specific activity of the two enzymes towards casein was 81,585 and 110,000 nmol 32P incorporated/hr/mg of enzyme. Protein kinase N1 comprised one polypeptide of Mr 35,000 which underwent phosphorylation in the presence of Mg2+ + ATP. Protein kinase N2 comprised two polypeptides Mr 40,000 and 30,000 of which only the Mr 30,000 polypeptide was autophosphorylated. Both enzymes were active towards casein, phosvitin, dephosphophosvitin, spermine-binding protein, and non-histone proteins in vitro. Little activity was detected towards histones. Both enzymes were stimulated by 150-200 mM NaCl. MgCl2 requirement varied with the protein substrate but was between 2-4 mM for both enzymes. With dephosphophosvitin as substrate, the apparent Km for ATP for N1 protein kinase was 0.01 mM. GTP did not replace ATP in this reaction. Protein kinase N2 was active in the presence of ATP or GTP. The apparent Km was 0.01 mM for ATP, but 0.1 mM for GTP. PMID- 3025030 TI - Purification of cytosolic cAMP-independent protein kinases from rat ventral prostate. AB - Two cAMP-independent protein kinases were purified from rat ventral-prostate and liver cytosol, and were designated PK-C1 and PK-C2 to distinguish them from the nuclear protein kinases described in the preceding paper. The yield of the prostate enzymes was about 5% each, and about 10% each for the liver enzymes. The average fold purification of the prostatic enzymes was 1892 and 3176 for protein kinase C1 and C2, respectively. Their average respective specific activity towards casein was 40,111 and 67,340 nmol 32P incorporated/hr per mg of enzyme protein. protein kinase C1 comprised one polypeptide of Mr 39,000 which underwent phosphorylation in the presence of Mg2+ + ATP. Protein kinase C2 comprised three polypeptides of Mr 41,000; 38,000; 26,000. Of these only the Mr 26,000 polypeptide was autophosphorylated. The Mg2+ requirement for protein kinase C1 and C2 was between 1 and 4 mM depending on the nature of the protein substrate. Both enzymes were stimulated by 100-200 mM NaCl. Km for ATP for C1 and C2 kinases was 0.01 mM; GTP could be used only by protein kinase C2 but with a markedly lower affinity. The enzymes were active towards casein, phosvitin, dephosphophosvitin, and spermine-binding protein in vitro, but demonstrated little activity towards histones. Despite several similarities in these general properties of cytosolic protein kinases C1 and C2 with those of nuclear protein kinases N1 and N2, a number of differences are also noted. PMID- 3025031 TI - Myosin light chain phosphorylation in contraction and relaxation of intact rat thoracic aorta. AB - Myosin light chain phosphorylation in intact rat thoracic aorta was elevated during contraction induced by 0.3 microM norepinephrine, but was not maintained. Addition of 0.5 microM sodium nitroprusside to norepinephrine treated rat aorta strips led to elevation of cyclic GMP levels, relaxation of tension, and dephosphorylation of myosin light chain. Depletion of extracellular calcium or addition of calmodulin antagonists trifluoperazine and W7 diminished the contraction and phosphorylation of myosin light chain by norepinephrine, but did not prevent dephosphorylation by sodium nitroprusside or the elevated levels of cyclic GMP. Isoproterenol, 8-bromo cyclic GMP, and dibutyryl cyclic AMP all caused dephosphorylation of myosin light chain and induced relaxation during the period of development of tone. Eight other proteins had increased phosphorylation following norepinephrine treatment and one protein had less phosphorylation. The different proteins phosphorylated by norepinephrine showed varying degrees of sensitivity to Ca2+-free solution and to the calmodulin antagonists. The pattern of protein phosphorylation caused by sodium nitroprusside was best mimicked by 8 bromo cyclic GMP, rather than isoproterenol and dibutyryl cyclic AMP. These proteins were, generally, unaffected by Ca2+-free solution and the calmodulin antagonists. The present observations support the hypothesis that vasodilators inhibit tone development through myosin light chain dephosphorylation. Furthermore, the nitrovasodilators act through elevation of cyclic GMP and phosphorylation of proteins by cyclic GMP-dependent protein kinase. PMID- 3025032 TI - Cyclic AMP changes in the component cells of Graafian follicles: possible influences on maturation in the follicle-enclosed oocytes of hamsters. AB - Mature antral follicles were removed from the ovaries of pregnant mare serum gonadotropin (PMSG)-primed hamsters at proestrus prior to the LH surge. Following various incubation times with either LH (ovine) or FSH (rat), cAMP levels were determined in whole follicles, cumulus-oocyte complexes (COCs), and zona-intact or zona-free oocytes. LH produced a dose- and time-dependent change in follicle cAMP but had a minimal effect on the COCs and caused no change in cAMP in zona free oocytes. By contrast, rFSH stimulated a small rise in follicular cAMP but significantly increased levels in COCs and zona-free oocytes. In a second series of experiments follicles were exposed for short periods to various additives after which they were washed and returned to hormone-free medium for a 6-hr total incubation period. LH (1 microgram/ml) initiated maturation in follicle-enclosed oocytes after a 5- to 15-min exposure period while groups incubated with 100 ng/ml required 60 min. FSH did not stimulate maturation after a 60-min exposure and when combined with 1 microgram or 100 ng/ml of LH negated the maturational effects seen with LH alone. It was postulated that the reason that lower concentrations of LH did not stimulate maturation following short-term incubations was due to an insufficient rise in cAMP. However, neither dbcAMP nor forskolin augmented the capacity of LH to initiate maturation following short term exposure. By contrast dbcGMP and the guanylate cyclase activator, sodium nitroprusside (NP) did augment the maturation-inducing effects of LH. NP + LH raised cGMP concentrations in the follicle and oocyte and decreased follicular cAMP at 30 and 120 min. The results of this study indicate that the component cells within a follicle respond selectively with cAMP changes, depending on the gonadotropin, in a variable time- and dose-dependent manner. While LH is the more potent activator of cAMP in whole follicles, cAMP levels in the cumulus oophorus and oocyte show the greatest increase following exposure to FSH. LH was the more potent initiator of maturation, possibly through its effects on the mural granulosa cells. FSH appears to exert a more inhibitory role which may be due in part to elevated cAMP levels and/or a putitative inhibitor in the COC and oocyte. PMID- 3025033 TI - Isolation and clonal analysis of satellite cells from chicken pectoralis muscle. AB - Satellite cells, liberated from the breast muscle of young adult chickens by sequential treatment with collagenase and trypsin, were fractionated by Percoll density centrifugation to remove myofibril fragments and cell debris which otherwise heavily contaminate the preparation. This procedure allowed direct measurements of cell yields (1.5-4 X 10(5) cells/g tissue), plating efficiencies (27-40%) and identification of single cells in culture. In mass cultures, satellite cells gave rise to myotubes on the fifth day, and the progeny of these cells were sequentially passaged several times without losing myogenic traits. In clonal studies, over 90% of the satellite cells gave rise to large clones of which more than 99% were myogenic as demonstrated by the appearance of myotubes. The results obtained with satellite cells differ from observations made using embryonic muscle cell preparation from chicks. In the embryonic system massive formation of myotubes was observed following the third day of culture; sequential subculturing led to overgrowth of fibroblast-like cells following the first passage; and cells gave rise to both small myogenic clones (up to 16 terminally differentiated cells per clone) and non-myogenic clones in addition to large myogenic clones. We conclude that the isolated satellite cells represent a homogeneous cell population and reside in a stem cell compartment. PMID- 3025034 TI - Synthesis and ubiquitination of histones during myogenesis. AB - One and two-dimensional polyacrylamide gel electrophoresis have revealed that cultures of postmitotic (G0) chicken skeletal myotube cells synthesize significant but reduced quantities of histone proteins as compared to their proliferating myoblast precursors. In addition, modulation of variant synthesis within the histone H2A and H3 classes may accompany myotube formation. That the histone bands contain no nonhistone contaminants was shown by exclusion of [3H]tryptophan. It is unlikely that these results reflect synthesis of histone by contaminating replicating cells, since a single treatment with cytosine arabinoside at the time of fusion effectively removed unfused cells while suppressing synthesis of DNA in the myotube cultures. The relatively sparse incorporation of label by major variants of the H2A class in dividing myoblasts was shown to be caused by heterogeneity due to phosphorylation and extensive ubiquitination, which decline at the time of myotube formation. As determined by quantitative Western-blotting, dividing myoblasts and myotubes contain an average of 1.0 and 0.4 molecules of ubiquitinated H2A (uH2A), respectively, per 10 nucleosomes. PMID- 3025035 TI - Reduction in in vivo testing at the National Veterinary Services Laboratories of the United States Department of Agriculture. AB - Efforts are being made at the National Veterinary Services Laboratories to reduce in vivo testing of USDA licensed veterinary vaccines. A hemagglutination test for determining potency of killed parvovirus vaccine is currently being used for canine and swine adjuvanted and nonadjuvanted products; a serum neutralization inhibition test (SNIT) is being developed for potency testing of killed adjuvanted infectious bovine rhinotracheitis (IBR), bovine virus diarrhea (BVD) and parainfluenza (PI3) vaccines: and a tissue culture titration method for live avian encephalomyelitis virus vaccine is being pursued as a replacement for the old hatch-out chick embryo titration method. Difficulties in separating the antigen from oil emulsion products are preventing significant advances in developing in vitro testing procedures for poultry killed-virus vaccines. PMID- 3025036 TI - The use of serum neutralizing antibody assay for the determination of the potency of foot and mouth disease (FMD) vaccines in cattle. AB - The Permanent Commission of O.I.E. on foot and mouth disease has proposed a series of potency standards for foot and mouth disease vaccines. No totally in vitro assay has yet been developed to satisfy these requirements, and most Control Authorities require a potency assay to be carried out in cattle using challenge with virulent virus. There are various reasons why challenge tests should be replaced by serological tests, but a stumbling block to this is the requirement that the 50% protective antibody level (PA50) is valid only for a specific combination of vaccine virus seed lot with cattle virus seed lot in a laboratory. The results of challenge and antibody tests for 38 U.K. Control Authority vaccine batch tests were analysed to examine the correlation between potencies measured by the two methods. The correlation between the methods was high and the proportion of misclassification of batches was low. It is suggested that the results provide a good basis for future use of the serological method, but that it is still sometimes necessary to carry out challenge tests. PMID- 3025037 TI - Reduction of animal usage in the control of foot-and-mouth disease vaccines. AB - FMD vaccines were evaluated by comparing the results of a series of assays made on virus suspensions and on the final product, to mean values derived from a data base containing information on more than 300 accepted vaccine lots. The potency was estimated from the average percentage characterizing the vaccine, using the general mean in vivo protective dose, also provided by the data base. The correlation between the in vitro estimated potency and the in vivo tested potency as made routinely, was relatively low. However the results obtained by both methods were always in good agreement. The estimation method appeared sufficient for the evaluation of the potency of FMD vaccines, at the stage of the production. This method could allow an important reduction in the animal usage for the control of FMD vaccines. PMID- 3025038 TI - Development of in vitro tests for detection of extraneous agents. AB - The use of animals in tests to detect extraneous agents is not only undesirable from the ethical viewpoint but also because of the expense and length of time involved in such tests. We have carried out tests on a variety of potential contaminating avian pathogens to determine whether tests in chicks offer any advantage over tests in embryos or cell cultures. In many cases, but not all, in vitro tests were shown to be more sensitive. The use fluorescent antibody or enzyme-linked assays serves to enhance the sensitivity of the tests. In the future it may be possible to adapt techniques such as nucleic acid hybridisation to the detection of extraneous agents. PMID- 3025039 TI - Dot immunobinding assay of viral antigen and antibodies. AB - A dot-immunobinding assay for the rapid and specific detection of viral antibody as well as the identification of a virus is described. Based on the use of nitrocellulose membranes for the adsorption of viral protein, this test may be used to detect the presence of virus antibody or identify an isolate within 4 to 6 hours. Use of goat anti-human IgG-alkaline phosphatase followed by a naphthol AS-MX-phosphate: Fast Red substrate permits visual detection of a positive reaction. The use of psoralen inactivated herpes B virus permits the incorporation of this virus into the battery of test antigens. PMID- 3025040 TI - A comparison of titration methods for live avian encephalomyelitis virus vaccines. AB - Factors associated with the indirect fluorescent antibody test used for the titration of avian encephalomyelitis virus (AEV) in chick embryo brain cell cultures were examined for their influence on virus replication. It was found that virus should be inoculated onto semi-confluent cell cultures and adsorbed for two hours at room temperature. The cells should then be examined for fluorescence after five days' incubation. Using these conditions, the cell culture assay was compared with the embryo and chick assays for its ability to estimate the virus content of live commercial AEV vaccines. In most cases titres obtained by the chick assay were slightly, but not significantly, higher than those obtained in the cell culture assay, although the reliability of the chick assay was, at times, questionable. In all cases titres obtained in the embryo assay were low. It is recommended that the cell culture assay be adopted as the method of choice for titrating AEV vaccines because it is rapid, reproducible, specific and greatly reduces the requirement for experimental animals. PMID- 3025041 TI - A semi-automated system for the assessment of toxicity to cultured mammalian cells based on detection of changes in staining properties. AB - We have established a semi-automated microtiter-based system for the quantification of dye binding to cultured eukaryotic cells. This system has been applied to the quantitation of toxic activities that disrupt cell monolayers and their neutralization. We have used this background as a basis for developing a detection and characterization system for activities that do not cause such gross toxicity. A prototype system has been established based on three staining procedures which in broad terms assess cellular dehydrogenase activity, and protein, DNA, and RNA content. The activity of several agents affecting cyclic nucleotide metabolism, including cholera toxin, on the staining properties of exposed monolayers has been assessed. Several new categories of cellular response are readily discernible in this latter system indicating that biological activities may be identified on the basis of the pattern of such responses. Since microtiter based systems show considerable potential for automation, it is suggested that the further development of this approach could offer a realistic prospect for numerous forms of toxicity testing on an industrial scale. PMID- 3025042 TI - Effects of fasting, feeding, and insulin on enhancing effect of GTP on cAMP binding in rat hepatic cytosol. AB - GTP, in physiologic concentration, enhances the binding of cAMP to a protein in the hepatic cytosol that may be the regulatory subunit of protein kinase II. Ingestion of carbohydrate suppresses hepatic gluconeogenesis and glycogenolysis, two processes that are stimulated by cAMP. In this study, we have examined the possibility that carbohydrate inhibits these processes partly by decreasing the sensitivity of the GTP-responsive cAMP-binding protein to the effect of GTP. We found that 100 muM GTP was much less effective in enhancing cAMP binding in the hepatic cytosol of rats given 15% glucose for 2 days than in the cytosol of fasted rats [21 +/- 3% (mean +/- SE) increase vs. 67 +/- 6%, P less than .01]. Corresponding results were noted in diethylaminoethyl (DEAE)-cellulose extracts of the hepatic cytosol of these rats. GTP stimulation of cAMP binding was also diminished in the hepatic cytosol of diabetic rats treated for 7 days with insulin compared with that of untreated diabetic rats (29 +/- 10 vs. 81 +/- 11% increase, P less than .01), but this could have been due to increased food intake in the treated rats. We conclude that GTP stimulation of hepatic cAMP binding is decreased in the carbohydrate-fed state and that this effect may be mediated by the increase in plasma insulin induced by carbohydrate. Our observations suggest that some of the cellular effects of cAMP may be regulated by modulation of the stimulatory effect of GTP on the GTP-responsive cAMP-binding protein. PMID- 3025043 TI - Effect of aldose reductase inhibitor ONO 2235 on reduced sympathetic nervous system activity and peripheral nerve disorders in STZ-induced diabetic rats. AB - To test the hypothesis that inhibitors of aldose reductase, a key enzyme for polyol synthesis, prevent the decrease of sympathetic nervous system (SNS) activity and improve motor nerve conduction velocity (MNCV) through the reduction of sorbitol accumulation in streptozocin (STZ)-induced diabetic rats, norepinephrine (NE) turnover (reliable indicator of SNS activity in the interscapular brown adipose tissue (IBAT), heart, and pancreas), and MNCV were measured in untreated STZ-induced diabetic rats, STZ-induced diabetic rats treated with an aldose reductase inhibitor (ARI) (ONO 2235, 50 mg X kg-1 X day-1, orally), STZ-induced diabetic rats treated with insulin therapy, control rats, and control rats treated with ARI. As expected, results from studies with the inhibition of NE biosynthesis with alpha-methyl-p-tyrosine or radiolabeled NE to measure NE turnover demonstrated significant reductions in SNS activity in IBAT, heart, and pancreas of untreated STZ-induced diabetic rats, compared with those in control rats. MNCV determined with the tail nerve was significantly reduced in untreated STZ-induced diabetic rats, compared with that of the controls. Both daily ARI treatment and insulin therapy in STZ-induced diabetic rats prevented partially but significantly the decrease of NE turnover in IBAT, heart, and pancreas, ameliorated MNCV, and reduced sorbitol accumulation in the nerve tissue and red blood cells. ARI treatment in control rats had no effect on NE turnover, MNCV, or sorbitol content. These results suggest that STZ-induced diabetic rats had not only motor neuropathy but also sympathetic nervous dysfunction and that ARI treatment might prevent these as well as insulin therapy does through the reduction of sorbitol accumulation. PMID- 3025044 TI - Presence of complement-dependent cytotoxic activity against clonally-derived rat islet tumour cells in sera from type 1 (insulin-dependent) diabetic patients and control subjects. AB - Heat-inactivated sera from newly diagnosed Type 1 (insulin-dependent) diabetic patients and control subjects were tested for the presence of antibodies to islet cell surface antigens by means of a sensitive immunofluorescent, microcytotoxicity assay using two clones of a rat islet cell tumour as antigens. Complement-dependent cytotoxicity was found in 74% of diabetic patient sera and 87% of control sera, and there were no significant differences in titres between diabetic patients and control subjects. A minority of the sera from both patients and controls were cytotoxic for only one of the two clones, suggesting the presence of multiple antigen-antibody systems. Preadsorptions of the sera with rat liver powder, sheep erythrocytes, and/or protein A-conjugated agarose beads were inconsistently effective in decreasing levels of lytic activity in control sera. It is concluded that more information is required concerning the antigens of rat islet cells and islet cell cytotoxic factors present in normal sera before such cells and assays can be reliably used for the detection of islet cell surface antibodies. PMID- 3025046 TI - Production and turnover of cAMP signals by prestalk and prespore cells in Dictyostelium discoideum cell aggregates. AB - Dictyostelium discoideum prestalk cells and prespore cells from migrating slugs and culminating cell aggregates were isolated by Percoll density centrifugation. Several activities relevant to the generation, detection, and turnover of extracellular cyclic AMP (cAMP) signals were determined. It was found that: the two cell types have the same basal adenylate cyclase activity; prespore cells and prestalk cells are able to relay the extracellular cAMP signal equally well; intact prestalk cells show a threefold higher cAMP phosphodiesterase activity on the cell surface than prespore cells, whereas their cytosolic activity is the same; intact prestalk cells bind three to four times more cAMP than prespore cells; no large differences in cAMP metabolism and detection were observed between cells derived from migrating slugs and culminating aggregates. The results are discussed in relation to the possible morphogenetic role of extracellular cAMP in Dictyostelium cell aggregates. On the basis of the properties of the isolated cells we assume that a gradient of extracellular cAMP exists in Dictyostelium aggregates. This gradient appears to be involved in the formation and stabilization of the prestalk-prespore cell pattern. PMID- 3025045 TI - Reduction of erythrocyte (Na+-K+)ATPase activity in type 1 (insulin-dependent) diabetic subjects and its activation by homologous plasma. AB - The (Na+-K+)ATPase and (Mg2+)ATPase activities of erythrocyte membranes of Type 1 (insulin-dependent) diabetic patients were found to be significantly reduced compared to matched controls (p less than 0.005). On the contrary, erythrocyte Na+ and K+ contents were similar in diabetic patients and in normal subjects. When erythrocyte membranes from diabetic patients were incubated with their own plasma, a significant increase was observed in sodium-potassium ATPase activity (p less than 0.005), whereas (Mg2+)ATPase activity was not affected. The plasma stimulatory effect showed saturation kinetics. Maximum average stimulation was 96% (+/- 21.3). A similar stimulation pattern, although more limited in extent (maximum 48.3% +/- 12.2), was found when erythrocyte membranes from normal subjects were incubated with diabetic plasma. Normal plasma exhibited a modest stimulatory effect on erythrocyte (Na+-K+)ATPase activity. Similar stimulatory effects by diabetic plasma were observed on a (Na+-K+) ATPase preparation from beef heart. It is proposed that diabetic plasma contains a specific (Na+ K+)ATPase activator in a higher concentration than normal plasma. This may explain why a normal cellular electrolyte content was found in diabetic erythrocytes in spite of a reduced Na+-K+ pump activity. Purification experiments indicate that the plasma activator is a protein with a molecular weight greater than 50,000. Both the (Na+-K+)ATPase activity and the stimulatory effect of diabetic plasma were not influenced by the metabolic control, since they did not correlate significantly with fasting blood glucose and daily insulin dosage. Moreover, no correlation was found with duration of diabetes or age at diagnosis of diabetes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3025047 TI - Control of laminin synthesis during differentiation of F9 embryonal carcinoma cells. A study using cDNA clones complementary to the mRNA species for the A, B1 and B2 subunits. AB - Molecular clones complementary to the mRNA species for the A, B1 and B2 chains of murine laminin were identified by hybrid-selection and in vitro translation. Northern blot analysis demonstrated that the three clones, p59 (A), p2 (B1) and p16 (B2) hybridized to mRNA species 9.8, 6.0, and 8.0 kb in length, respectively. The three clones were used as probes to monitor the steady-state levels of laminin mRNA species during differentiation of F9 embryonal carcinoma cells induced by treatment with retinoic acid and dibutyryl cyclic AMP. The steady state levels of the three mRNA species appeared to increase in a coordinate manner. Undetectable levels at the beginning of induction were followed by a dramatic increase in the levels of the three mRNA species between 48 and 72 h. The kinetics parallel the increase in laminin synthesis and the striking morphological changes previously reported. PMID- 3025048 TI - Differentiation of embryonic mouse GH cells and ACTH cells in vitro. AB - The ability of cells that produce growth hormone (GH) and cells that produce adrenocorticotropic hormone (ACTH) to differentiate in various culture media was analyzed by means of ultrastructural immunocytochemistry on 13-day embryonic mouse pituitaries that were maintained in organ culture for 3-11 days. At the time of culture, relatively undifferentiated nongranulated or poorly granulated cells that were unreactive with anti-growth-hormone serum (anti-GH) and anti adrenocorticotropic-hormone serum (anti-ACTH) were present in the pituitary. After 10-11 days in culture, immunoreactive GH cells were obtained only in media that were supplemented with cortisol, whereas ACTH cells were obtained in all media tested, including Medium 199 alone. In cortisol-supplemented media, the GH cells showed ultrastructural features typical of those that occur in vivo, and anti-GH immunoreactivity was obtained after as little as 3 days in culture, i.e., at a stage comparable to that which occurs in vivo. The results indicate that mouse GH cells are capable of differentiating in Medium 199 supplemented only with cortisol, without the addition of fetal calf serum or insulin; cortisol therefore appears to be an essential component of the embryonic milieu for the production of GH-secretory granules. PMID- 3025049 TI - [Adenylate cyclase activity--after stimulation of beta-receptors with isoproterenol--in relation to the presence or absence of monocytes in a population of mononucleated cells]. PMID- 3025050 TI - [Presence of DNA sequences of hepatitis B virus in a fibrolamellar carcinoma of the liver]. AB - We report the case of a twenty-one-year old woman using oral contraceptives who underwent partial hepatectomy for fibrolamellar carcinoma. Anti-HBs and anti-HBc antibodies were present in the serum and chronic hepatitis was associated with the carcinoma. Study of the tumor with DNA hybridization techniques showed the presence of integrated DNA sequences of the hepatitis B virus in the tumor cells. This observation underlines the possible interactions of different factors in hepatic carcinogenesis, and suggests that estrogens may possibly act as a cocarcinogenic factor associated with viral infection. PMID- 3025051 TI - Dietary fibers in the preparation of the bowel for diagnostic barium enema. AB - The effect on colon cleanness of adding dietary fiber or placebo to a standard bowel preparation scheme before diagnostic barium enemas was tested in a randomized, double-blind study including 120 consecutive outpatients admitted for diagnostic barium enemas. Colon cleanness was scored as satisfactory, intermediate, or unsatisfactory. No case of unsatisfactory cleansing appeared in the group given fiber. Mean score was slightly better in the fiber group but the difference was not statistically significant. Dietary fiber does not have a negative effect on colon cleansing but may instead have a beneficial effect, which is possibly more pronounced in patients with constipation. PMID- 3025053 TI - [Cytomegalovirus infection after bone marrow transplantation]. PMID- 3025052 TI - Vasoactive intestinal polypeptide actions on the guinea pig intestinal mucosa during neural stimulation. AB - Electrical stimulation of the mucosal innervation of the guinea pig ileum results in an increase in chloride secretion that is mediated in part by excitation of muscarinic cholinergic receptors on enterocytes. This study investigated the involvement of vasoactive intestinal peptide in the cholinergic and noncholinergic phases of the secretory response evoked by electrical stimulation of submucosal neurons in the guinea pig ileum. Flat sheets of ileum set up in Ussing flux chambers responded to exogenous vasoactive intestinal peptide by an increase in baseline short-circuit current which was reduced by furosemide and by vasoactive intestinal peptide antiserum. When submucosal neurons were electrically stimulated, a biphasic change in short-circuit current was evoked. Vasoactive intestinal peptide, forskolin, and isobutylmethylxanthine enhanced the cholinergic portion of the response, whereas the antiserum prevented or reduced the effects of the peptide but not of forskolin. In the presence of atropine to eliminate the cholinergically mediated response, vasoactive intestinal peptide reduced the noncholinergic phase of the response and its action was prevented by the antiserum. Vasoactive intestinal peptide enhanced the increase in short circuit current evoked by the muscarinic agonist bethanechol. These results demonstrate that vasoactive intestinal peptide and other substances that stimulate secretion by increasing cyclic 3',5'-adenosine monophosphate levels in enterocytes potentiate the calcium-dependent, cholinergic phase of the chloride secretory response evoked by neural stimulation of the guinea pig ileum. No evidence was found for vasoactive intestinal peptide as the mediator of the noncholinergic phase of the response. PMID- 3025054 TI - [Study of the functional activity of the membrane Na+, K+-ATPase of preserved erythrocytes using a radionuclide method]. PMID- 3025055 TI - [Cytochrome oxidase in bone marrow cells of the rabbit after intravenous administration of hydroxyethylated glycerin]. PMID- 3025056 TI - Inverse modulation of extracellular Na+- and K+-activities by ascorbate or methylene blue. AB - On analyzing the mechanisms of the internal environment type redox regulation of physiological processes it was observed on frog rectus muscles that during acetylcholine contractures methylene blue pretreatment inhibited, but ascorbate pretreatment enhanced the slow transient changes of extracellular Na+-activity. At the same time, these modifications were inverse for K+-transients. Because k strophantoside was capable of influencing these effects radically it seems highly plausible to assume that the principal site of action of these modulations is the inhibitory impact of methylene blue, while the enhancing effect of ascorbate on (Na+ + K+)-ATPase may likely be explained on redox basis. PMID- 3025057 TI - A comparative study of microscopic and macroscopic parameters of lipid bilayers. PMID- 3025058 TI - Stable non-mutator stocks of maize have sequences homologous to the Mu1 transposable element. AB - Mutator stocks of maize produce mutants at many loci at rates 20- to 50-fold above spontaneous levels. Current evidence suggests that this high mutation rate is mediated by an active transposable element system, Mu. Members of this transposable element family are found in approximately 10-60 copies in Mutator stocks. We report here an initial characterization of previously undetected sequences homologous to Mu elements in eight non-Mutator inbred lines and varieties of maize that have a normal low mutation rate. All stocks have approximately 40 copies of sequences homologous only to the terminal repeat and show weak homology to an internal probe. In addition, several of the stocks contain an intact Mu element. One intact Mu element and two terminal-specific clones have been isolated from one non-Mutator line, B37. The cloned sequences have been used to demonstrate that in genomic DNA the intact element, termed Mu1.4B37, is modified, such that restriction sites in its termini are not accessible to cleavage by the HinfI restriction enzyme. This modification is similar to that observed in Mutator lines that have lost activity. We hypothesize that the DNA modification of the Mu-like element may contribute to the lack of Mutator activity in B37. PMID- 3025060 TI - [Comparative analysis of the organization of the NPL-1 plasmid controlling naphthalene oxidation in Pseudomonas putida and its derivatives]. AB - The paper contains the data on the structure of NPL-1 plasmid controlling naphthalene biodegradation. The plasmid which pertains to the P-9 incompatibility group is transferrable conjugatively and is maintained stably within a wide range of gram-negative bacteria. The analysis of mutants and transposon derivatives of the plasmid made it possible to localize nah-genes in a DNA fragment, 23 kb in size. An inverted DNA segment of 4.2 kb was discovered which may participate in the regulation of nah-genes expression. The other peculiar features of NPL-1 were found distinguishing it from NAH and NAH7 plasmids described in literature. PMID- 3025059 TI - Thymidine utilization by tut mutants and facile cloning of mutant alleles by plasmid conversion in S. cerevisiae. AB - Plasmid pJM81 contains a Herpes simplex virus thymidine kinase (TK) gene that is expressed in yeast. Cells containing the plasmid utilize thymidine (TdR) and the analogue 5-bromodeoxyuridine (BUdR) for specific incorporation into DNA. TdR auxotrophs, harboring plasmid pJM81 and a mutation in the yeast gene TMP1 require high concentrations of TdR (300 micrograms/ml) to support normal growth rates and the wild-type mitochondrial genome (rho+) cannot be maintained. We have identified a yeast gene, TUT1, in which recessive mutations allow efficient utilization of lower concentrations of TdR. Strains containing the mutations tmp1 and tut1, as well as plasmid pJM81, form colonies at 2 micrograms/ml TdR, grow at nearly normal rates and maintain the rho+ genome at 50 micrograms/ml TdR. These strains can be used to radiolabel DNA specifically and to synchronize DNA replication by TdR starvation. In addition, the substitution of BUdR for TdR allows the selective killing of DNA-synthesizing cells by 310-nm irradiation and allows the separation of replicated and unreplicated forms of DNA by CsCl equilibrium density banding. We also describe a unique, generally applicable system for cloning mutant alleles that exploits the fact that Tk+ yeast cells are sensitive to 5-fluorodeoxyuridine (FUdR) and that gene conversions can occur between a yeast chromosome and a TK-containing plasmid. PMID- 3025061 TI - [Mutations predetermined by the primary structure of DNA]. AB - This study is concerned with an experimental verification of hypotheses postulating the involvement of self-complementary nucleotide sequences in the formation of deletions and insertions. It was suggested that deletions can arise in the regions of self-complementary nucleotide sequences, which allows the formation of the hairpin structures in a single-stranded DNA, arising during excision repair. These hairpin structures can be eliminated by nucleases or during DNA replication. Insertions can arise as a result of homologous recombination, when a migrating DNA strand contains a self-complementary sequence which forms hairpin structure. Model experiments were carried out with the pBR322 plasmid. A plasmid DNA with premutational damage in the palindrome-containing region was constructed by in vitro dimethylsulfate modification of one strand of EcoRI-BamHI restriction fragment. The plasmid was used for transformation of Escherichia coli. Restriction mapping and nucleotide analysis of the mutant DNAs demonstrated that they all contained deletions. The end points of the deletions coincide with the palindrome. To model homologous recombination, a plasmid with D loop was constructed. A single-stranded DNA fragment containing palindrome forming a hairpin structure was introduced into the plasmid DNA and covalently fixed in the complex. When E. coli cells were transfected with this DNA, plasmid mutants containing insertions predetermined by palindromic structure arose. The evolutionary role of mutations predetermined by primary DNA structure is discussed. PMID- 3025062 TI - [Interaction of mobile elements P and mdg3 in Drosophila melanogaster: genetic aspects]. AB - It was found earlier that two unstable sn mutants isolated from natural populations are connected with insertion of mobile element mdg3 into the 7D1-2 region where singed gene (1-21.0) is localised. From two original sn mutants, a series of unstable sn alleles, both mutant and normal for phenotype, was extracted. Then we studied, how they change the mutation rate in germinal and somatic cells of different hybrids with pi 2 stock having P cytotype and active P elements in the chromosomes. Addition of P chromosomes, independently of the background of cytoplasm, proved to reduce the sn instability. The level of sn mutability was decreased with increasing the dose of P chromosomes. It is suggested that mutation events are caused by transposition of mdg3 and that both mdg3 and P elements compete for the same cellular factor, capable of activation of transposition process. PMID- 3025063 TI - Cell death in the development of the human retina: phagocytosis of pyknotic and apoptotic bodies by retinal cells. AB - Apoptosis is a natural form of cell death and has features in common with aspects of cell deletion observed in the course of human retinal development. In this report, we describe the occurrence of apoptotic cells in various layers of the developing retina. Pyknotic residues were observed within phagosomes of neighbouring retinal cells. Our observations imply that most of the debris resulting from cell death is taken up by adjacent tissue cells rather than by mononuclear phagocyte series cells (macrophages) during early stages of human retinal development. PMID- 3025064 TI - [Work conditions and diseases of the shoulder girdle among women engaged in plastering and painting]. PMID- 3025065 TI - Topics and controversies in enteral nutrition. Introduction. PMID- 3025066 TI - Mixed mesodermal tumor of the uterus (RJ-984): case report and in vitro study. AB - This report describes the establishment of a cell line of a human uterine mixed mesodermal tumor. The tumor of origin derived from a hysterectomy specimen, has been maintained for 14 months in vitro and continues to grow as an established cell line. The original tumor as well as the cell line exhibited no estrogen receptors. alpha-Fetoprotein was not detected in the cultured cells or in the spent culture medium. Karyotyping revealed 46 XX chromosome complement with a balanced 11-16 translocation. This is the first documentation of such a chromosome abnormality in a genital tract carcinoma. Steroids inhibited cell growth at high (10.0 micrograms/ml) concentrations. This cell line continues to be studied and further characterized. The cell line is readily available for study of this aggressive human neoplasm. PMID- 3025067 TI - Integration of transposon Tn10 into phage L (Salmonella typhimurium). AB - Transposon Tn10 was transposed into phage L (Salmonella typhimurium) from F'ts114lac+zzf::Tn10 plasmid of strain TT629 (Chumely et al. 1979). Phage L with the insertion Tn10 (L::Tn10-8) was isolated in the form of a prophage in the lysogenic strain S. typhimurium LT2-18 (L::Tn10-8), in which it can be induced with UV light. The phage induced in this way is defective; however, it forms plaques at a multiplicity of infection (moi) greater than one and transduces the tetracycline-resistance determinant to tetracycline-sensitive cells. Analysis of its DNA by restriction endodeoxyribonucleases revealed insertion of the intact transposon Tn10 of 9300 bp in the E fragment, formed during the action of EcoRI, at a distance of 16,800 bp from the pac site. PMID- 3025068 TI - Kaluresis independent K-homeostasis: glucagon and B receptor blockade in pancreatectomized dogs. AB - Seventy minutes post pancreatectomy, in dogs that are K loaded - made abruptly hyperkalemic and "life threatened" - by infusion with 2 mEgKC1/kg-/hr until prelethal ECG changes of hyperkalemic cardiotoxicity appear, a kaluresis independent K homeostatic mechanism transfers about 2/3 of administered K to intracellular fluid. Treatment of K loaded pancreatectomized dogs with glucagon or a B receptor blockading dosage of propranolol does not alter the proportion transferred, but treatment with glucagon and propranolol reduces it. It appears that in pancx dogs there is a reciprocal relation between insulin and B receptor mediated K transfer and that glucagon is involved in activity of the kaluresis independent K homeostatic mechanism. PMID- 3025069 TI - Cell-mediated cytotoxicity in hepatitis A virus infection. AB - We studied cell-mediated cytotoxicity to hepatitis A virus-infected cells in seven patients with acute type A hepatitis and two controls. Skin fibroblast cultures obtained from the skin biopsies of seven patients after acute hepatitis A virus infection and from two persons without history of current or past hepatitis A virus infection were inoculated with hepatitis A virus. Infection of fibroblast cultures always resulted in an inapparent, persistent infection with production and release of infectious hepatitis A virus. Peripheral blood lymphocytes were collected from the same patients at different times after onset of icterus and were stored in liquid nitrogen. Cytolytic activity of peripheral blood lymphocytes was determined by a microcytotoxicity assay using autologous 51Cr-labeled hepatitis A virus-infected and uninfected target cells. Cytotoxic peripheral blood lymphocytes capable of lysing autologous hepatitis A virus infected skin fibroblasts were detected in all patients with hepatitis A but were not demonstrable in the controls without antibodies against hepatitis A virus. The clinical course of the hepatitis A virus infection was normal in five patients; and in these patients, cytolytic activity of peripheral blood lymphocytes against hepatitis A virus-infected autologous targets peaked 2 to 3 weeks after onset of icterus. A clinically protracted form of the disease with persistent elevation of aminotransferases for at least 5 months after onset was present in two patients. In these cases, the highest cytolytic activity was demonstrated in peripheral blood lymphocytes collected 8 to 12 weeks after onset of icterus.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3025070 TI - Brainstem auditory evoked potentials in hepatic encephalopathy. AB - Brainstem auditory evoked potentials were obtained from 30 patients primarily with viral hepatitis to determine the functional status of the brainstem in hepatic encephalopathy. The patients were divided into four groups: Group 1 with compensated hepatic diseases; Group 2 with hepatic failure but no hepatic encephalopathy; Group 3 with Grade 1 or 2 hepatic encephalopathy, and Group 4 with Grade 3 or 4 hepatic encephalopathy. The brainstem auditory evoked potential central conduction times (I-V interval) of four patient groups were not significantly different from that of the age-matched control group. The present data suggest that the brainstem auditory pathways remain undisturbed in hepatic encephalopathy. PMID- 3025071 TI - Monoclonal antibodies against antigens expressed on human hepatocellular carcinoma cells. AB - Monoclonal antibodies with selectivity for human hepatoma cell lines were produced by immunizing BALB/c mice with human hepatoma cell lines, HA22T/VGH or Hep 3B, and fusing sensitized mouse spleen cells with mouse myeloma cells. Two monoclonal antibodies recognizing antigens present only on human hepatoma cell lines were investigated. The monoclonal antibody IB1 was found to react with 3 of 9 hepatoma cell lines. Monoclonal antibody 9B2 reacted with all nine hepatoma cell lines. None of the other 20 cell lines tested was bound by IB1 and 9B2. The immunoperoxidase staining of monoclonal antibodies on frozen sections of paired hepatoma and normal liver tissues from the same individuals were studied. Antibody IB1 reacted with 3 of 13 hepatoma tissues, but with none of the normal liver and other tissues, and antibody 9B2 was reactive with antigens appearing on the bile canalicular domain of hepatoma and normal liver tissues. The antibody 9B2 stained no normal tissues with the exception of proximal tubules of kidney. Radioimmunoprecipitation tests identified two antigens reacting with 9B2. The major antigen had an apparent molecular weight of 140,000 and a minor one of 130,000. Therefore, antibody IB1 seems to be specific for antigens present on a group of human hepatoma cells and may be useful for classification and diagnosis of human hepatomas. Antibody 9B2 is quite specific to human liver cells and may be used to provide clues for the characterization of tumor cell lines, identification of metastatic tumors with hepatocytic origin, and study of the structure and function of bile canaliculi. PMID- 3025072 TI - Intrauterine latent herpes simplex virus infection: I. Spontaneous abortion. AB - Herpes simplex virus (HSV, probably type 2) antigen was detected in nonpregnant and pregnant endometria, placentae, umbilical cords, and neonatal tissues (companion paper) by avidin-biotin complex immunohistochemical studies. HSV cytologic abnormalities were not detected in any of the 380 cases examined: included were specimens from therapeutic and spontaneous abortions (200 cases) and endometrial curettage (180 cases). The presence of inflammation was not correlated with HSV positivity. Endometrial HSV positivity was significantly correlated with normal late secretory phase (40 per cent of specimens positive), abnormal secretory phase (67 per cent positive), and therapeutic (33 per cent positive) versus spontaneous (26 per cent positive) abortions. Placental HSV positivity was significantly correlated with spontaneous (39 per cent positive) versus therapeutic (14 per cent positive) abortions and with blighted ova (67 per cent positive). No significant correlation was found between HSV positivity and a clinical history of oral or genital HSV infection in either the patient or the male partner. The data support the concept of a subclinical latent intrauterine endometrial HSV infection that is hormonally regulated and can produce transplacental infection of the embryo or fetus, with variable consequences. PMID- 3025073 TI - Intrauterine latent herpes simplex virus infection: II. Latent neonatal infection. AB - Herpes simplex virus (HSV, probably type 2) antigen has been detected in endometria and abortion tissue (companion paper) and in placentae, umbilical cords, and fetal and neonatal organs by avidin-biotin complex immunohistochemical studies. HSV cytologic abnormalities were not detected in any of the 12 normal and 64 abnormal cases analyzed, nor was HSV detected by culture or electron microscopy in selected cases. Antigen was present in single epithelial and, rarely, mesenchymal cells of various organs. Clinically unexplained fetal or neonatal problems associated with HSV antigen positivity included intrauterine death, fetal growth retardation, cystic brain degeneration, hydrops, interstitial pneumonitis, necrotizing enterocolitis, hepatitis, encephalitis, myocarditis, and renal failure. Maternal floor infarct of placenta and calcifying funisitis are the manifestations of intrauterine HSV infection in most cases. Maternal history of HSV infection was uncommon. It is concluded that intrauterine HSV infection may persist in the fetus and neonate in a latent fashion without cytologic abnormalities or detectable virus. This latent infection may be associated with intrauterine and neonatal death, organ damage, and neonatal disease. PMID- 3025074 TI - In situ hybridization detection of human papillomavirus DNAs and messenger RNAs in genital condylomas and a cervical carcinoma. AB - Routinely processed formalin-fixed paraffin-embedded sections from anogenital condyloma acuminatum and an invasive squamous cell carcinoma of the cervix were examined by in situ hybridization for the detection of human papillomavirus (HPV) DNAs and messenger RNAs. Asymmetric, single-stranded, tritium-labeled RNA probes for both the coding and the nonsense strands of HPVs 6, 11, 16, 18, and 31 were hybridized and washed under stringent conditions and detected by autoradiography. Type-specific HPV DNAs were detected with specific nuclear localization, while HPV messenger RNAs gave much higher signals and had clear-cut cytoplasmic localization. Cross-hybridization was observed only with closely related viruses. The level of signal obtained seemed to be linked to the degree of cellular differentiation, with koilocytotic cells labeling the most heavily. However, messenger RNA could be detected in even relatively undifferentiated cells within areas of dysplasia and invasive carcinoma. In situ hybridization is a sensitive and specific method for investigation of the dynamic interplay of papillomavirus replication and gene expression, cellular differentiation, and neoplastic transformation. PMID- 3025075 TI - Atypical endocrine tumors of the lung: a histologic, ultrastructural, and clinical study of 19 cases. AB - Lung cancers are divided by light microscopic criteria into several categories, but only two categories are recognized clinically--small cell and non-small cell carcinomas. Transmission electron microscopy has revealed unexpected complexity within each category, blurring the distinctions between them. The present study was undertaken to determine the incidence of dense-core, neuroendocrine-type granules in lung tumors diagnosed by light microscopy as non-small cell carcinomas, i.e., atypical endocrine tumors, and the clinical significance of their identification. Of 205 consecutive primary and metastatic lung cancers, 19 (9 per cent) diagnosed as non-small cell carcinomas by light microscopy were seen to contain neuroendocrine-type granules by electron microscopy and thus were reclassified as atypical endocrine tumors of the lung. Staining with silver stains, periodic acid-Schiff (PAS), PAS with diastase digestion, and mucicarmine was positive in 18, 15, 14, and eight of the 19 cases, respectively. Electron microscopy revealed glandular differentiation in 12 cases and tonofilaments in eight cases, although none of the tumors met the criteria for identification as squamous cell carcinomas. Clinically, the cancers appeared to resemble non-small cell carcinoma more closely than small cell carcinoma. Median survival (12 months) and response to combination chemotherapy (22 per cent) were in the range reported for non-small cell carcinoma. There were no complete responses, despite the use in some cases of regimens active against small cell carcinoma. However, one patient, the only one to date so treated, had a dramatic response to streptozotocin/5-fluorouracil, suggesting that, as in metastatic carcinoid, this combination may have value in the treatment of atypical endocrine tumors of the lung. PMID- 3025076 TI - An elongated segment of DNA observed between two human alpha globin genes. AB - Detailed restriction enzyme analysis of the DNA from a Chinese female showed that one of her chromosomes had a greater than 17.5 kb deletion of DNA, including the psi alpha, alpha 2, and alpha 1 globin genes, which is present in many Southeast Asians with an alpha-thalassemia-1 chromosome. Her "normal" chromosome had the expected cluster of alpha-like globin genes (5'-zeta-psi zeta-psi alpha-alpha 2 alpha 1-3'), but the segment of DNA between the two alpha globin genes was elongated by some 0.5-0.7 kb. Analyses of various restriction sites suggested that this normal variant of the human alpha globin gene complex is due to a crossover between a normal chromosome with (alpha alpha) and a chromosome with an alpha-thalassemia-2 (-alpha 3.7) and an -alpha 2 alpha 1-hybrid gene. PMID- 3025077 TI - New regional localisations for HAGH and PGP on human chromosome 16. AB - The chromosomal locations for the electrophoretic markers hydroxyacyl glutathione hydrolase (HAGH) and phosphoglycolate phosphatase (PGP) were examined using a human-mouse hybrid panel of chromosome 16. The assignment for HAGH was confirmed to chromosome 16 using a cell line with chromosome 16 as the only human chromosome. Both HAGH and PGP were present only in cell lines containing human 16p13. This localisation for PGP indirectly places the tightly linked genes for the alpha-globin cluster and adult polycystic kidney disease on 16p13. PMID- 3025079 TI - Phosphatidate phosphohydrolase from Culex pipiens fatigans. PMID- 3025078 TI - Population structure of eastern Sicily. AB - A sample of 465 persons from Eastern Sicily was studied for 11 red-cell enzymes, namely GLO, GPT, EsD, PGP, PGD, Dia, AcP, PGM, SOD, CAI and CAII. The allele frequencies were compared with those of other Italian populations and showed that the island is homogeneous with the mainland for these systems. The rate of heterozygosity was studied as a function of interparental distance; although high (0.77) the correlation did not reach significance. PMID- 3025080 TI - Effect of dietary protein on cholesterol lowering action of blackgram fiber. PMID- 3025081 TI - EB virus induction is associated with B-cell maturation. AB - EB virus genome-carrying B-cell lines have been double-stained for B-cell activation or maturation antigens and viral antigens using indirect immunofluorescence. The vast majority of cells express activation antigens, whereas the plasma cell antigen PC1 is present on only a few cells. Staining for this antigen doubles with staining for EB viral capsid antigen. Thus it marks cells in the virus replicative cycle, and these cells are negative for the EB virus nuclear antigen EBNA. We suggest that within EB virus genome-carrying B cell lines the maturation of a few cells to the stage of expression of plasma cell markers leads to the loss of EBNA expression and consequent loss of the immortalized state of the cell. This would allow completion of the lytic cycle with the production of virus particles and cell death. The implications of these findings are discussed. PMID- 3025082 TI - [Primary mucinous carcinoma of the skin]. PMID- 3025083 TI - Recombination hot spots within the I region of the mouse H-2 complex map to the E beta and E alpha genes. AB - Recombinant mouse strains with crossovers in the I region of the H-2 major histocompatibility complex were examined by restriction fragment analysis for the presence of polymorphic restriction sites within the E beta and E alpha genes. Nine recombinant mouse strains were shown to have crossed over within a 5 kb DNA segment that contains the large intron between the second and third exons of the E beta gene. These results are in accord with previous studies mapping a recombination hot spot within this gene. Seven recombinant mouse strains between the p and k haplotypes were shown to have crossed over in a 6 kb segment within the E alpha gene. These results show the existence of a recombination hot spot within the E alpha gene. Comparison of the H-2 haplotypes involved in these two recombination hot spots suggests that a specific DNA sequence in b, s, f, and q haplotypes may act to promote recombination in the E beta gene and a specific DNA sequence in the p haplotype may act to promote recombination in the E alpha gene. PMID- 3025084 TI - Extensive deletions in the Q region of the mouse major histocompatibility complex. AB - By use of Southern blot analyses and low copy number probes, the fine structure of the Q region of the mouse major histocompatibility complex was studied in more detail. With a probe recognizing the even-numbered genes Q4, Q6, and Q8, it was evident that Q4 and/or the regions flanking Q4 are polymorphic, whereas Q6 and Q8, and their flanking regions are nonpolymorphic. Perhaps the most noteworthy finding is that at least two strain haplotypes, H-2k and H-2f, possessed extensive deletions in the Q region. The most striking deletion was found in the H-2f haplotype, where the Q1 through Q9 genes appear to be missing. Because of these extensive deletions the functional importance of the Q region is questioned. PMID- 3025085 TI - Polymorphic and autoreactive H-2-specific monoclonal antibody isolated after injections of syngeneic Sendai virus-coated lymphocytes. AB - An H-2-specific monoclonal antibody (mAb Q-1) was obtained from B10.Q (H-2q) mice injected with syngeneic Sendai virus-coated cells. The IgM monoclonal antibody recognizes the public determinant H-2.25 shared by H-2k (Kk) and H-2r haplotypes and cross-reacts with H-2d, H-2s, H-2p, and H-2q cells, the latter being the haplotype of the challenged B-cell donor. The binding of mAb Q-1 to H-2d, H-2s, H 2q, and H-2p cells was lower than to H-2k and H-2r and of decreasing affinity but could be clearly distinguished from the negative reactions with H-2b and H-2f cells. MAb Q-1 distinguishes between Sendai virus-coated and uncoated lymphocytes only cells with low-affinity binding. On virus-coated or infected (H-2p, H-2q, H 2d, H-2s) cells lysis was stronger than on normal lymphocytes. We interpret the enhanced lysis of Sendai virus-positive cells by mAb Q-1 to be due to recognition of a modified exposure of public H-2 determinants induced by Sendai virus. PMID- 3025087 TI - DNA cross-linking by chloroethylating agents. PMID- 3025086 TI - Characterization of non-T effector cell subpopulations involved in production of human alpha interferon. AB - The phenotype of the human effector leukocyte subset involved in production of alpha interferon was examined using positive and negative selection techniques including murine monoclonal antibodies. The data suggest that the effector cell responsible for the elaboration of human alpha interferon is surface immunoglobulin positive, lacks either the E-rosette receptor or any monocyte determinants and is also negative for expression of the surface antigens identified by Leu-10 and Leu-11b monoclonal antibodies. Neither monocytes nor polymorphonuclear leukocytes were shown to play a role in regulation or production of alpha interferon. The data further imply, however, that the Leu-7+, large granular lymphocyte subpopulation may play a critical role in regulating human alpha interferon production by surface immunoglobulin positive B cell effectors. PMID- 3025088 TI - Renin inhibitors. PMID- 3025089 TI - Effects of angiotensin inhibition and renal denervation in two-kidney, one clip hypertensive rats. AB - Neural and angiotensin-mediated influences that alter hemodynamic and excretory behavior of the nonclipped kidney of two-kidney, one clip hypertensive rats were assessed by sequential acute surgical denervation of the nonclipped kidney and intravenous infusion of converting enzyme inhibitor (SQ 20881), 3 mg/kg X hr. Normal and two-kidney, one clip hypertensive rats (0.2-mm silver clip on the right renal artery 3-4 weeks before study) were prepared to allow study of each kidney. Mean arterial blood pressure of two-kidney, one clip hypertensive rats fell significantly from control values of 149 +/- 6 to 135 +/- 6 mm Hg after denervation of the nonclipped kidney. Despite this decrease in arterial pressure, the nonclipped kidney exhibited significant increases in glomerular filtration rate (from 1.00 +/- 0.08 to 1.24 +/- 0.08 ml/min), sodium excretion (from 88 +/- 39 to 777 +/- 207 nEq/min), fractional sodium excretion (from 0.06 +/- 0.02 to 0.54 +/- 0.14%), and urine flow rate (from 3.7 +/- 0.5 to 8.2 +/- 1.1 microliter/min). A significant decrease in glomerular filtration rate (from 1.12 +/- 0.07 to 0.85 +/- 0.08 ml/min) with no change in excretory function was observed for the clipped kidney following denervation of the nonclipped kidney. Intravenous addition of converting enzyme inhibitor significantly increased renal blood flow (from 7.0 +/- 1.3 to 10.6 +/- 1.5 ml/min) and sodium excretion (from 777 +/- 207 to 1384 +/- 425 nEq/min) for the nonclipped kidney; blood pressure decreased from 135 +/- 6 to 123 +/- 4 mm Hg, and renal vascular resistance decreased significantly (from 22 +/- 3 to 13 +/- 2 mm Hg X min/ml).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3025091 TI - Neuropeptide regulation of interleukin-1 activities. Capacity of alpha-melanocyte stimulating hormone to inhibit interleukin-1-inducible responses in vivo and in vitro exhibits target cell selectivity. AB - alpha-Melanocyte stimulating hormone (alpha-MSH), a 13 amino acid neuropeptide produced by the pituitary gland, was found to markedly inhibit the capacity of exogenously administered interleukin-1 (IL-1) to stimulate the enhanced synthesis of acute-phase proteins and induce neutrophilia in vivo. The administration of ACTH or glucocorticosteroids lacked most of these direct IL-1 inhibitory properties. Therefore, in addition to the previously reported antipyretic action of alpha-MSH, this hormone can also inhibit two other known IL-1 sensitive cellular targets in vivo. Further, alpha-MSH was incapable of modifying the comitogenic influence of IL-1 on murine thymocytes or on an IL-1 responsive T cell line. These findings suggest a target cell specificity to the IL-1 inhibitory activities of alpha-MSH and fail to support the hypothesis that alpha MSH functions through competitive inhibition of specific cellular receptors for IL-1. PMID- 3025090 TI - Clostridial collagenase. A chemoattractant for human neutrophils. AB - Leukocyte chemoattractants markedly alter the morphology and membrane functions of leukocytes. Bacterial collagenase causes a change in cell shape similar to that seen with the leukocyte chemoattractant, f-Met-Leu-Phe, and also promotes capping of concanavalin A. Human neutrophils in suspension or adherent to cover glasses were exposed to clostridial collagenase (10-250 units/ml) for up to 30 min at 37 degrees C and then fixed. Collagenase (125 units/ml) caused more than 85% of PMNs to assume an asymmetric or motile morphology even in the presence of 1% gelatin or 10 mg/ml bovine serum albumin. Trypsin alone (0.01-1%) did not induce a shape change. A similar morphology was seen in some untreated PMNs (less than 5% of all cells) and is characteristic of f-Met-Leu-Phe-treated cells (more than 90%). Collagenase inhibitors (i.e., reduced glutathione, cysteine, and acid soluble collagen), however, prevented the shape change induced by collagenase but not by f-Met-Leu-Phe. At 4 degrees C, fluorescein-Con A (20 micrograms/ml) bound uniformly to both untreated and collagenase-treated cells. Upon further incubation at 37 degrees C, Con A was internalized over the entire cell periphery of the rounded, untreated cells but on collagenase-treated PMNs was rapidly gathered into a cap overlying the uropod or protuberant region of cytoplasm where it was subsequently internalized. Checkerboard Boyden chamber assays showed clostridial collagenase to be chemokinetic and chemotactic for human PMNs. In receptor binding experiments, the clostridial collagenase preparation competed poorly with [125I]formylhexapeptide for binding to PMN formylpeptide receptors (less than 15% reduction in binding at 200 units/ml collagenase). Thus, collagenase does not seem to interact strongly with the neutrophil formylpeptide receptor and may stimulate PMN motility by interacting at an altogether different site. PMID- 3025092 TI - Effects of dietary fish oil supplementation on polymorphonuclear leukocyte inflammatory potential. AB - Polymorphonuclear leukocytes (PMNLs) are an important contributor to inflammation and are thus a part of the pathophysiology of many human diseases. We assessed the effect of fish oil on PMNL inflammatory potential by measuring chemiluminescence and superoxide production before and after six weeks of daily cod liver oil ingestion by healthy volunteers. Phagocytosing PMNLs demonstrated a 27% decrease in chemiluminescence (P less than 0.05) and a 64% decrease in superoxide production (P less than 0.01), following the cod liver oil supplementation. Analysis of PMNL and platelet fatty acids revealed the appearance of eicosapentaenoic acid and a significant decrease in arachidonic acid in both types of cells. PMID- 3025093 TI - Carrageenan stimulates reduction of nitroblue tetrazolium by human neutrophils without membrane depolarization, myeloperoxidase secretion, or increased oxygen consumption. AB - Carrageenan, a sulfated polyanionic polysaccharide, is commonly used to induce inflammation in experimental animals, and this model is used to screen for the effectiveness of antiinflammatory drugs. Carrageenan-induced inflammation has been attributed to a variety of autocoids including histamine, bradykinin, complement, superoxide, and prostaglandins. This study examines the effects of carrageenan on human PMN in a serum-free system. Carrageenan was found to stimulate the reduction of NBT by PMNs without stimulating membrane depolarization, oxygen consumption, H2O2 production, or myeloperoxidase secretion. Carrageenan stimulates a heat-labile, NBT-reducing system which is unassociated with the usual stimulus-response coupling seen with other PMN activators such as PMA, FMLP, and zymosan. PMID- 3025094 TI - Altered function of synovial fluid granulocytes in patients with acute inflammatory arthritis: evidence for activation of neutrophils and its mediation by a factor present in synovial fluid. AB - In rheumatoid arthritis (RA) a chronic inflammatory state exists in which the synovial fluid is periodically filled with large numbers of polymorphonuclear leukocytes (PMNs). Oxygen radicals produced by these cells have been implicated as mediators of tissue damage and may be directly involved in the pathogenesis of RA. We examined the production of oxygen radicals by synovial fluid PMNs (SF PMNs) and peripheral blood PMNs (PB-PMNs) by measuring chemiluminescence (CL) as well as superoxide anion (O2-) release. Increased spontaneous CL in the presence of luminol and increased CL in response to phorbol myristate acetate (PMA) was observed in SF-PMNs when compared to PB-PMNs. When zymosan was used as the stimulus in the absence of luminol, a slightly lower CL response was observed in SF-PMNs as compared to PB-PMNs. No significant differences were observed in the generation of O2- generation with any stimulus. Preincubation of normal PB-PMNs in 10% synovial fluid enhanced the luminol-dependent spontaneous and PMA stimulated CL as well as zymosan-stimulated CL. When O2- release from normal PB PMNs pretreated with 10% synovial fluid was compared to untreated controls, enhancement of spontaneous O2- release was observed. PMA- and zymosan-stimulated responses did not differ significantly from controls. Increased spontaneous and PMA-stimulated release of myeloperoxidase (MPO) was also observed in normal PB PMNs pretreated with synovial fluid. These findings may explain the increased luminol-dependent CL since this type of CL requires the presence of MPO. Our findings suggest that the enhanced chemiluminescence observed in normal PMNs treated with synovial fluids may be related to increases in spontaneous O2- generation and myeloperoxidase release. Increased MPO release may account for enhanced CL observed in SF-PMNs. PMID- 3025095 TI - Persistent bactericidal defect in neutrophils from a young woman who recovered from toxic shock syndrome. AB - We have previously found transient menstruation-associated abnormalities in the in vitro bactericidal function of neutrophils from females who have recovered from toxic shock syndrome (TSS). We now report the case of a young woman who has also recovered from TSS, but who has a persistent, non-menstruation-associated defect in the ability of her neutrophils to kill Staphylococcus aureus in vitro. PMID- 3025096 TI - Increased incidence of menstruation-associated bactericidal defects in neutrophils from women who have recovered from toxic shock syndrome. AB - There is a growing suspicion that a host abnormality may contribute to the pathogenesis of toxic shock syndrome (TSS). We found that females (5 of 5) who had recovered from TSS had transient, menstruation-associated decreases (greater than or equal to 9%) in the ability of their neutrophils to kill Staphylococcus aureus. 502A in vitro more often (P = 0.040 by Fisher's exact test) than non-TSS affected control subjects (5 of 12). In addition, the average decrease in bactericidal activity in neutrophils obtained during menstruation from recovered TSS patients was 30 +/- 9% compared to 7 +/- 7% for neutrophils from non-TSS affected control subjects. The results are consistent with the possibility that transient menstruation-associated decreases in neutrophil bactericidal function may indicate susceptibility and/or contribute to the development of TSS. PMID- 3025097 TI - Mechanism of leukotriene generation in polymorphonuclear leukocytes by staphylococcal alpha-toxin. AB - The effects of staphylococcal alpha-toxin on arachidonic acid metabolism in rabbit polymorphonuclear leukocytes (PMNs) were investigated and compared with those of the ionophore A23187 and the chemotactic tripeptide formylmethionyl leucyl-phenylalanine (fMLP). Sublytic amounts of alpha-toxin stimulated the release of leukotriene B4 (LTB4) in PMNs in a dose-dependent manner. The toxin was several times more potent than fMLP but was not as effective as the ionophore. Preincubation of the toxin with neutralizing antibodies abolished the effect. Extracellular calcium was strictly required for eliciting LTB4 generation. Verapamil, a calcium channel blocker, inhibited fMLP-mediated LTB4 generation but had no effect on alpha-toxin- or A23187-exposed PMNs. Agents such as trifluoperazine and N-6(aminohexyl)-5-chloro-1-naphthalene sulfonamid that interfered with calmodulin activity, however, inhibited LTB4 generation in all cases. One minute after the addition of alpha-toxin, PMNs exhibited a severalfold enhancement in passive permeability to 45Ca2+. In addition, these cells became permeable to sucrose but not to inulin or dextran. The influx pattern was consistent with the previous observation that alpha-toxin creates discrete transmembrane channels in erythrocytes with an effective internal diameter of 2 to 3 nm. The results suggest that alpha-toxin triggers the arachidonic acid pathway in PMNs by facilitating calcium influx into the cells, possibly via transmembrane toxin pores that serve as calcium gates. Generation of arachidonic acid metabolites in PMNs by sublytic amounts of alpha-toxin may represent an important cellular reaction that generally occurs during infections with Staphylococcus aureus. PMID- 3025099 TI - Induction of mucosal immunoglobulin A immune response by preparations of Neisseria gonorrhoeae porin proteins. AB - The aim of this study was to develop an immunization scheme appropriate for the induction of an immunoglobulin A (IgA) response against Neisseria gonorrhoeae at the mucosae. For several reasons, the major outer membrane protein of N. gonorrhoeae (gonococcal PI) is attractive as a component of a gonococcal vaccine. Purified PI obtained from strain B2 was used in Zwittergent 3-14 for immunization. Rats received the antigen subcutaneously, intraintestinally, or directly in Peyer's patches with or without the adjuvant N-acetylmuramyl dipeptide (MDP) or AlPO4. The immune response was studied in situ by a newly developed antigen-specific three-step immunoperoxidase method, whereas specific antibodies in the serum were measured by an enzyme-linked immunosorbent assay procedure. Subcutaneous immunizations triggered peripheral lymphoid organs, whereas intraintestinal and intra-Peyer's patch immunizations triggered mucosa associated lymphoid organs. This was reflected not only in the lymph nodes involved, popliteal versus mandibular and mesenteric lymph nodes, but also in the isotypes of the produced anti-PI antibodies, IgG versus IgA. The adjuvants AlPO4 and MDP appeared to act differently during subcutaneous and intraintestinal immunizations. AlPO4 augmented subcutaneous immune responses, whereas MDP had no effects. In contrast, intraintestinal immune responses increased most with the adjuvant MDP. In summary, we concluded that PI is capable of inducing a mucosa associated IgA response when administered intraintestinally and that this response can be augmented by the adjuvant MDP. PMID- 3025098 TI - Protein sources of heme for Haemophilus influenzae. AB - Although Haemophilus influenzae requires heme for growth, the source of heme during invasive infections is not known. We compared heme, lactoperoxidase, catalase, cytochrome c, myoglobin, and hemoglobin as sources of heme for growth in defined media. The minimum concentration of heme permitting unrestricted growth of strain E1a, an H. influenzae type b isolate from cerebrospinal fluid, was 0.02 micrograms/ml. Using molar equivalents of heme as lactoperoxidase, catalase, cytochrome c, myoglobin, and hemoglobin, we determined that myoglobin and hemoglobin permitted unrestricted growth at this concentration. To determine the ability of host defenses to sequester heme from H. influenzae, we used affinity chromatography to purify human haptoglobin and hemopexin, serum proteins which bind hemoglobin and heme. Plate assays revealed that 12 strains of H. influenzae acquired heme from hemoglobin, hemoglobin-haptoglobin, heme-hemopexin, and heme-albumin. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of outer membrane proteins of strain E1a grown in heme-replete and heme-restricted conditions revealed a heme-repressible outer membrane protein with an apparent molecular mass of 38 kilodaltons. These results demonstrated that, unlike Escherichia coli, H. influenzae may acquire heme from hemoglobin-haptoglobin. H. influenzae also may acquire heme from hemopexin and albumin, which have not been previously investigated. The role of outer membrane proteins in the acquisition of heme is not yet clear. PMID- 3025100 TI - Histoplasma capsulatum fails to trigger release of superoxide from macrophages. AB - The yeast form of the dimorphic fungus Histoplasma capsulatum survives within macrophages after phagocytosis. To do so, it must avoid, inhibit, or resist a variety of toxic oxygen metabolites. Using ferricytochrome c reduction to assay superoxide release, we examined the response of mouse macrophages to the yeast form of various H. capsulatum strains. Doses of zymosan as low as 20 particles per macrophage elicited superoxide, whereas H. capsulatum failed to induce superoxide even at 160 yeast cells per macrophage. This phenomenon was observed with two virulent strains of H. capsulatum (G217B and G186A) and with an avirulent variant of G186A. Over a 15- to 150-min observation period, zymosan stimulated increasing reduction of ferricytochrome c, but H. capsulatum did not. When added concurrently with zymosan, H. capsulatum had no effect on superoxide production. Therefore, H. capsulatum was unable either to inactivate the oxygen radical or inhibit host cell superoxide response to other competent stimuli. Enzymatically generated superoxide reduced ferricytochrome c even in the presence of H. capsulatum, again implying that the organism does not readily inactivate superoxide. This experiment also demonstrated that the yeast did not interfere with the assay used. Thus, rather than inhibiting superoxide generation or inactivating the anion, H. capsulatum yeast cells appear to avoid the toxic effects of superoxide by failing to trigger its release. PMID- 3025103 TI - Compression neuropathies: medical aspects and legal implications. PMID- 3025102 TI - Anti-delta antibody in primary hepatocellular carcinoma patients in the Gizan area of Saudi Arabia. PMID- 3025101 TI - Effects of cancer chemotherapy on the human aerobic oropharyngeal flora. AB - Since various agents used in cancer chemotherapy exhibit antimicrobial activity in vitro, we performed sequential quantitative cultures of saline gargles obtained from patients receiving cancer chemotherapy to determine if such chemotherapy alters the composition of the aerobic oropharyngeal flora. When we compared results of cultures obtained from 12 patients just before and at various times after receiving courses of cancer chemotherapy, we observed small, though significant reductions in the numbers of total bacteria, alpha-hemolytic streptococci and inhibitory streptococci two to seven days after courses of chemotherapy. A concomitant increase in the percentage of patients colonized by gram-negative bacilli occurred. Of the chemotherapeutic agents used to treat our subjects, only doxorubicin exhibited antimicrobial activity in vitro. All four alpha-hemolytic streptococci, but none of the seven strains of gram-negative bacilli examined, were inhibited by doxorubicin at concentrations of less than or equal to 12.5 mg/l. Doxorubicin had a modest enhancing effect on in vitro adherence of gram-negative bacilli to human embryonic lung cells. These data suggest that cancer chemotherapy might play a role in colonization of cancer patients by gram-negative bacilli by creating a microbiologic vacuum conducive to such colonization. In this way, cancer chemotherapy might contribute to the high incidence of gram-negative bacillary pneumonia among patients with malignant neoplasms. PMID- 3025104 TI - Plant-derived diterpene esters enhance HTLV-I-induced colony formation of lymphocytes in co-culture. AB - The addition to culture dishes of 10-50 ng/ml of the essential diterpene ester of Sapium sebiferum, 12-O-hexadecanoylphorbol-13-acetate (HPA), increased colony formation of normal peripheral blood lymphocytes co-cultured with gamma irradiated HTLV-I-producing HUT 102 cells. The cells in the stimulated colonies showed an approximately 3-fold increase in the expression of interleukin-2 (IL-2) receptors and a 1.5- to 2.0-fold increase in human T-lymphotropic virus type- I (HTLV-I) p19-positive cells. This biological potency was analogous to that induced by the most potent tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) and stronger than that of 12-O-hexadecanoyl-16-hydroxyphorbol-13-acetate (HHPA) isolated from Aleurites fordii. PMID- 3025105 TI - Retinoblastoma cell differentiation in culture. AB - Human retinoblastoma cells were grown in tissue culture and their differentiation into Flexner-Wintersteiner rosettes was investigated. This process of photoreceptor cell differentiation was only observed in primary cultures and subsequent cell passages of tumors which showed these structures in vivo. Rosettes formed spontaneously within 5-9 d after plating of the tumor cells. Under optimal conditions in certain tumor cell strains, up to 80% of the cultured cell aggregates contained one or more differentiated Flexner-Wintersteiner rosettes. Exposure of retinoblastoma cells to RA or dcAMP did not alter the number of rosettes in culture. Retinoblastoma cells within rosettes continued to synthesize DNA, and mitotic figures were frequently observed in histological sections. Ultrastructural analysis of rosettes formed in vitro showed many of the characteristics described in those found in vivo including a polarized shape, established cell junctions (Zonula adherens), extensive accumulation of mitochondria and microtubules in the apical part of the cells, numerous basal bodies and centrioles as well as cilia typical of the mature photoreceptor cell. Lamellated stacks of membranes were also found and their nature is discussed. PMID- 3025106 TI - EBV-negative and -positive Burkitt cell lines variably express receptors for B cell activation and differentiation. AB - The expression of receptors for proliferation and differentiation factors was analyzed by indirect immunofluorescence on 29 Burkitt lymphoma (BL) cell lines previously classified into 3 groups on the basis of their reactivity with 8 monoclonal antibodies (MAbs), including anti-CALLA, BL13 and TU1. BL13 and HB5 antibodies recognize different epitopes of the EBV/CR2 receptors. The determinant recognized by BL13 has been previously shown to be expressed only on cell lines of the first two groups, supposed to derive from the germinal center and to be negative on a third group of lines of putative BM origin and established from sporadic cases of BL. In contrast, and as expected from its reactivity on normal B cells in the BM or in the lymph nodes, HB5 antibody reacts with all BL lines except one. The receptor for transferrin is expressed on the 29 lines. Two new MAbs, Bac-1 and B1H5, could recognize respectively receptors for BCGF1 and BCGF2. Bac-1 reacts with 15 of 17 BL lines belonging to the first two groups and 7 of 12 BL lines of the third group; 14 of 15 EBV + lines express Bac-1. No BL line expresses B1H5. The IL2 receptor is weakly expressed on 5 EBV + cell lines and one EBV (-) line. All delta are BCGF1-positive. The almost constant expression of BCGF1 receptor on EBV + cell lines is the only strict relation between the expression of receptors for growth factors and their characteristics (i.e. EBV association, translocation, ethnic origin and clinical presentation). The maturation stage or the origin of BL cell lines in relation to the expression of growth factor receptors and the functional significance of these receptors will be discussed. PMID- 3025107 TI - Epidemiological and environmental evidence of the health effects of exposure to erionite fibres: a four-year study in the Cappadocian region of Turkey. AB - An environmental and epidemiological study has been carried out in Central Cappadocia, Turkey, aiming at investigating the relationship between exposure to naturally occurring erionite fibres and the reported high incidence of malignant mesotheliomas. Airborne fibre levels are generally low but show a higher proportion of erionite fibres in the villages affected by malignant disease than in a control village. The same pattern is confirmed by analysis of the fibre content in lung tissues of sheep from several villages, both affected and unaffected by malignant disease. The 3 villages with the highest proportion of erionite fibres have high rates of malignant pleural mesothelioma, malignant peritoneal mesothelioma and lung cancer. No case of malignancy for the same sites has been reported during the study period from the control village. The relationships between these findings and their consistency with the results from experimental studies indicate erionite fibres as a carcinogenic agent, although some aspects of the exposure are not fully clarified. PMID- 3025108 TI - Reactive T cells in the immune repertoire: self-restricted and allo-restricted helper T-cell clones to Epstein-Barr virus. AB - Helper T-cell clones were generated by stimulation with autologous or allogeneic lymphoblastoid B cells (B-LCL) transformed by the Epstein-Barr virus (EBV). Some of these T-cell clones were allo-reactive and others were specific to EBV transformed B-LCL. Helper T-cell clones specific to EBV-transformed B-LCL were restricted either by class-I or by class-II HLA molecules of self. T-cell clones restricted by class-I HLA molecules were stained by OKT3 and OKT8 monoclonal antibodies (MAbs), whereas class-II-restricted clones stained with OKT3 and OKT4. Not all helper T-cell clones specific to EBV-transformed B-LCL were restricted to self: one clone restricted by allo-HLA antigen was established. This finding suggests that in humans, as in mice, some T cells in the T-cell repertoire can be allo-restricted. This allo restriction may represent cross-reactivity of T cells, whereby "self + X" equals "allo + Y." Activation of these cross-reacting T cells restricted by allogeneic HLA molecules during infectious mononucleosis will give a T-cell response which may appear unrestricted by self HLA molecules. This mechanism helps to explain, at least in part, the HLA unrestricted cytotoxicity to B-LCL observed in infectious mononucleosis. PMID- 3025109 TI - The differentiated form of nasopharyngeal carcinoma contains Epstein-Barr virus DNA. AB - Immunologic studies of Epstein-Barr virus (EBV) have implicated EBV in undifferentiated and partially differentiated, non-keratinizing nasopharyngeal carcinoma (NPC). Patients with the well-differentiated, keratinizing form of NPC have EBV serologic patterns similar to those of control populations. In addition, viral DNA has not been detected in the differentiated tumors using viral cRNA probes to DNA immobilized on filters. In this study we have tested for EBV DNA using recombinant DNA probes to Southern blots of DNA from 33 NPC specimens. The 24 undifferentiated and 4 partially differentiated specimens generally contained a relatively high number of EBV genome equivalents, while the 5 well differentiated NPC all contained detectable EBV, but at low copy number. The viral DNA from one of the well-differentiated specimens was cloned into a cosmid vector. Five recombinant clones representing the fused viral termini were obtained, indicating the presence of episomal, intracellular DNA in the tumor. These findings indicate that all histologic subsets of NPC contain EBV DNA. PMID- 3025110 TI - Hepatitis B virus, tobacco smoking and ethanol consumption in the etiology of hepatocellular carcinoma. AB - Tobacco smoking and alcohol drinking histories were obtained from 194 patients with hepatocellular carcinoma (HCC) and 456 hospital controls, and the results were analysed in conjunction with the results of serological determinations of hepatitis B surface antigen (HBsAg), antibody to HBsAg (anti-HBs) and antibody to hepatitis B core antigen (anti-HBc) in all subjects, as well as the presence or absence of cirrhosis in HCC patients. The relative risk (RR) of HCC (and 95% confidence interval) among HBsAg-positive subjects was 13.7 (8.0-23.5), whereas the excess risk among antibody-positive subjects was small and statistically non significant. In the presence of cirrhosis the RR for HBsAg-positive subjects was considerably higher (30.7 vs. 7.1 among HBsAg-positive subjects without cirrhosis) indicating that HBV may affect the development of HCC through at least two different and potentially multiplicative mechanisms (DNA integration and liver regeneration). Moderate ethanol consumption does not affect the risk of HCC, but there is a statistically significant and dose-dependent association between tobacco smoking and HBsAg-negative HCC. In most of the developed countries of Europe and North America, where the prevalence of HBsAg carrier state is very low and tobacco smoking very common, more cases of HCC may be due to tobacco smoking than to HBV, even though the RR for HCC is much higher among HBsAg carriers than among tobacco smokers. PMID- 3025112 TI - Alpha-radiation-induced amplification of integrated SV40 sequences is mediated by a trans-acting mechanism. AB - Treatment of Chinese hamster embryo cells with alpha irradiation (4 MeV, emitted by 241americium) induces a 15-fold amplification of integrated SV40 sequences. The extent of amplification depends on the dose of irradiation and on the presence of a functional T-antigen encoded by the SV40 A gene. The inducing signal can be transmitted to a non-irradiated nucleus following cell fusion. Amplification is therefore the result of some trans-acting process, and this could explain how SV40 amplification can occur after doses of alpha irradiation that are too low to cause direct DNA damage within the SV40 replicon. PMID- 3025111 TI - Association of BK virus with human brain tumors and tumors of pancreatic islets. AB - BK virus (BKV) DNA was detected by Southern blot hybridization in 19 out of 74 (25.6%) human brain tumors and in 4 out of 9 (44.4%) human tumors of pancreatic islets. BKV DNA was free, in an episomal state and generally in a low copy number (0.2 to 2 genome equivalents per cell). Only occasional tumors contained 10 to 20 genome copies per cell. Viral DNA sequences integrated into cellular DNA were not detected. A number of tumors expressed BKV-specific RNA and T antigen. By transfection of total tumor DNA into human embryonic fibroblasts, viruses with the biological and antigenic properties of BKV were rescued from 6 brain tumors and from 2 tumors of pancreatic islets. Restriction endonuclease mapping of the genomes of the rescued viruses showed that they differ from wild-type BKV. They are all similar to each other and to BKV-IR, a virus previously rescued from a human tumor of pancreatic islets, suggesting the possible association of a BKV variant with specific types of human neoplasms. The significance of the relationship of these BKV variants to human tumors and their possible etiologic role in human oncogenesis are discussed. PMID- 3025113 TI - Lung function improvement, tremor measurements and c-AMP determinations in a group of ten patients with asthmatic bronchitis after one week sustained release theophylline medication (theophylline plasma level +/- 10 mg/l) compared to one week placebo. AB - After one week theophylline therapy with a sustained release preparation (TheoDur 200 and 300 mg), a mean theophylline level of 10 mg/l was obtained in a group of ten patients with chronic asthmatic bronchitis with a mean dosage of 760 mg theophylline/24 h. A good improvement of the lung function was achieved, although only statistically significant for the mid maximal expiratory flow rate (MMEF) and the end expiratory specific airway conductance (sGaw) compared to placebo therapy. Tremor measurements and cyclic-AMP determinations showed a slight increase after theophylline therapy compared to placebo, although not statistically significant. PMID- 3025114 TI - Chagasic IgG binds and interacts with cardiac beta adrenoceptor-coupled adenylate cyclase system. AB - It has been previously shown that sera from chagasic patients have an antibody specific for beta adrenoceptors, independently of other tissue-reactive antibodies as the EVI (endocardium, blood vessels, interstitium) system, and it is highly specific to other heart diseases. In this paper we demonstrate that the IgG present in chagasic sera was able to bind to beta adrenoceptors of the heart and also to interact with the membrane bound adenylate cyclase complex, inducing stimulation of enzymatic activity. Moreover, this antibody stimulated contractile activity of guinea pig myocardium, that could be blocked by a specific beta adrenoceptor antagonist. Chagasic IgG inhibited the binding of (-)-(3H) dyhidroalprenolol to a beta-adrenergic receptor of purified guinea pig myocardial membranes behaving as non-competitive inhibitor. This IgG also exerted a non competitive inhibition upon the mechanical effect of exogenous norepinephrine. PMID- 3025115 TI - Synthesis of four diastereomeric enkephalins incorporating cyclopropyl phenylalanine. AB - Four diastereomeric D-Ala2, Leu5-enkephalins have been synthesized and their CD spectra and rat brain binding affinities determined. Only the peptides containing Z-cyclopropyl phenylalanine residues showed strong binding affinity. No correlation between CD spectra and bioactivity could be made. PMID- 3025116 TI - Intermolecular charge transfer involving tryptophan, tyrosine and three electron bonded intermediates derived from methionine. AB - Oxidation processes of radiation-generated three-electron-bonded intermediates derived from methionine Met2[S+...S] and Met[S...X] (X=Cl,Br) were investigated through reaction with tryptophan and tyrosine, using the optical pulse radiolysis method. Bimolecular rate constants have been measured for the reactions Met2[S+...S] with TrpH(k=3.8 x 10(8) dm3 mol-1 s-1 and k = 4.9 X 10(8) dm3 mol-1 s-1 at at ph 1 and 4.3, respectively) and Met2[S+...S] with tyrosine, k=3.8 x 10(7) dm3 mol-1 s-1. Rate constants for intermolecular transformation of Met[S...Br] and Met[S...Cl] into TrpH+. or Trp. were also estimated. They varied from 4.7 X 10(8) dm3 mol-1 s-1 (bromide species) to 1.0 X 10(9)dm3 mol-1 s-1 (chloride species). It has also been established azide radicals N-6, N.3 in contrast to dihalide radicals (X-2) do not form transients of Met[S...X] (X = N3) type. However, oxidation of N-3 by Met2[S+...S] occurs with a bimolecular rate constant of 2.8 X 10(8) dm3 mol-1 s-1. These results are discussed in the light of some equilibria which have been proposed earlier for methionine-halide systems. PMID- 3025117 TI - The reactions of the hydroxymethyl radical with 1,3-dimethyluracil and 1,3 dimethylthymine. AB - Hydroxymethyl radicals .CH2OH, generated by the radiolysis of methanol (0.5 mol dm-3) in N2O-saturated aqueous solutions, were reacted with 1,3-dimethyluracil or 1,3-dimethylthymine (10(-3) mol dm-3). The products were identified and their G values determined. It has been concluded that in 1,3-dimethyluracil .CH2OH attack occurs only at C(6) while in 1,3-dimethylthymine there is partitioning between addition (two-thirds) and H-abstraction from the C(5)-methyl group (one-third). A rate constant for CH2OH addition to 1,3-dimethyluracil of about 10(4) dm3 mol-1 s 1 is estimated. Complexities that may arise in the radiolysis of pyrimidines such as 1,3-dimethylthymine, apparently as a consequence of the formation of 5 alkylidenepyrimidines, are discussed. A value of 0.15 has been estimated for the disproportionation/combination ratio for the hydroxymethyl radical self termination reaction. PMID- 3025118 TI - Biological effectiveness of low energy protons. I. Survival of Chinese hamster cells. AB - The biological effectiveness of monoenergetic protons was investigated with the track-segment method. Protons were accelerated by a Tandem Van de Graaff accelerator and their final energies were 3.0 and 7.4 MeV. The biological system used was Chinese hamster V-79 cells and their survival ability following proton irradiation was investigated. Cobalt-60 gamma-rays were used as reference radiation to assess proton relative biological effectiveness (RBE). Survival curves were obtained for the gamma-ray and proton irradiations, and the relation S = exp (-alpha D-beta D2) was fitted to the data and the parameters alpha and beta were determined. The RBE values, calculated on the basis of the mean inactivation dose D and other pertinent parameters, were found to be 1.7 +/- 0.1 and 2.8 +/- 0.2 for 7.4 and 3.0 MeV protons, respectively. Comparisons were made with the results published by other investigators and it was concluded that in this low energy range the biological effectiveness increases substantially with decreasing proton energy. PMID- 3025119 TI - Purine and its analogues and radiation damage in Bacillus megaterium spores. AB - As an extension of results obtained from radiation studies on caffeine both in other laboratories and more recently in this laboratory using the bacterial spore as the test system, six compounds with chemical structures closely resembling that of caffeine were tested as radiation modifiers. Of these compounds, purine, adenine and hypoxanthine resembled caffeine in sensitizing spores to radiation, while theobromine, xanthine and theophylline did not. These responses are discussed in relation to the electron sequestration hypothesis of cellular sensitization to high-energy radiation. PMID- 3025120 TI - Binding experiment KD values and physiologically active GABA concentrations: an only apparent contradiction? AB - An apparent contradiction exists between GABAA receptor binding KD's (less than 10(-6) M) and GABA concentrations physiologically active in various systems (greater than 10(-6) M). Taking in account that synaptic cleft GABA action and removal should be rather quick (within 2 ms) we show that in principle there is no contradiction between low KD values (100-150 nM) and high physiologically active GABA concentrations (greater than 40 microM). PMID- 3025121 TI - Tourette syndrome: a case for noradrenergic and opiatergic mechanisms. PMID- 3025122 TI - Is excessive opioid activity relevant to the Korsakoff psychosis? PMID- 3025123 TI - The probable role of superoxide produced by blast cells in leukaemic cutaneous spreading. AB - Leukaemic blast cells from 12 patients with acute leukaemia were examined in order to study their capacity to produce anion superoxide in the absence and in the presence of phorbol-myristate acetate (PMA). Blast cells are able to mount an oxidative respiratory burst upon challenge with PMA, as demonstrated by superoxide release. The production of anion superoxide by blasts committed to monocytic differentiation might be an additional factor contributing to the tissue damage observed in leukaemic patients. PMID- 3025124 TI - Cardiovascular alterations in the rabbit infused with platelet activating factor (PAF): effect of kadsurenone, a PAF-receptor antagonist. AB - The ECG and haemodynamic alterations caused by the i.v. infusion of platelet activating factor (PAF) in the rabbit were studied after pretreatment with Kadsurenone, a specific PAF-receptor antagonist. Infusion of PAF at 0.8 microgram/kg in the rabbit caused important ECG changes, such as ST-segment depression and conduction arrhythmias, concomitantly with a marked reduction in left ventricular systolic pressure, mean arterial pressure and cardiac output, with a rise in total peripheral resistances and mean right atrial pressure. These physiological parameters became maximal at 75-120 seconds after PAF challenge, and returned to near prechallenge values within 25-60 minutes. Pretreatment with Kadsurenone, administered either intravenously (0.014 M, 1 ml/kg) or intraperitoneally (0.14 M, 1 ml/kg), exerted a quite complete protective effect in regard to the ECG changes and caused a significant reduction in the magnitude of all the haemodynamic alterations observed after intravenous infusion of PAF (0.8 microgram/kg). These results suggest that Kadsurenone is an effective inhibitor of PAF-induced cardiovascular changes in the rabbit, probably due to its competitive antagonism against PAF binding to specific receptors. PMID- 3025125 TI - Prevalence and distribution of spotted fever and typhus infections in Sierra Leone and Ivory Coast. AB - A serosurvey for evidence of rickettsial infections was conducted in the rural populations of several tropical rain forest areas in Sierra Leone and Ivory Coast. Seropositivity rates were surprisingly high in both countries, with more than 7% of the individuals in some districts having antibodies to spotted fever group rickettsiae. No significant difference was found in the overall prevalence of diagnostic antibody titers to spotted fever-group rickettsiae in Sierra Leone (5.3%) and Ivory Coast (6.2%). However, there was a significant difference (p less than 0.001) in the prevalence of diagnostic antibody titers to typhus rickettsiae in the two countries. There were no marked geographic differences within either country in overall prevalence of rickettsial infections, but there were possible area differences in specific seropositivity rates to typhus- and spotted fever-group rickettsiae in Sierra Leone. In both countries, age and sex differences were important in determining seropositivity, but there was no indication of an age-sex interaction. In Sierra Leone, 59 of the 80 positive sera (73.8%) were from persons age 15 or above (p less than 0.001), and 50 of the 80 (62.5%) were from males (p = 0.05). In Ivory Coast, 33 of the 37 positive sera (89.2%) were from the greater than or equal to 15-age group, and 28 of the 37 (75.7%) were from males (p less than 0.001 for both age and sex). The identification of specific areas endemic for these rickettsial diseases should facilitate the diagnosis and treatment of patients with rickettsial illnesses in West Africa. PMID- 3025126 TI - 'Milker's nodule' contracted from pox in water buffaloes. PMID- 3025127 TI - Plasmid analysis of simultaneous nosocomial outbreaks of methicillin-resistant Staphylococcus aureus. AB - A large outbreak of infections caused by methicillin and aminoglycoside resistant Staphylococcus aureus provided the opportunity to evaluate mechanisms of resistance and compare the usefulness of typing systems. Between January 1979 and December 1980, 63 patients developed infections with S aureus resistant to multiple antibiotics, including methicillin and tobramycin. All isolates had an identical antibiogram and were phage type 47/54/75/77/83A. Beginning in January 1981, a superimposed outbreak caused by S aureus of the same phage type but with a resistance pattern now including gentamicin occurred. The two strains contained different aminoglycoside inactivating enzymes. The initial strain contained a single plasmid of 21.5 mDa molecular weight, whereas the subsequent strain which had acquired gentamicin resistance contained this plasmid plus a heavier one of 33 mDa. Plasmid analysis complements the analysis of antibiograms and phage types and aids in defining epidemiologic patterns of transmission. PMID- 3025128 TI - [Significance of prostaglandins and leukotrienes in gastroenterology]. PMID- 3025129 TI - Inhibition of lens opacification in x-irradiated rats treated with WR-77913. AB - Radiation induced cataracts are models for studying mechanisms of lens opacification. WR-77913, S-3-(amino-2-hydroxypropyl) phosphorothioate (NCS 318809), has been identified as a radioprotective agent. Injection of WR-77913 (1160 mg/kg, i.p.) 15 to 30 min before exposure to 15.3 gray of x-irradiation inhibited rat lenses from developing radiation cataracts. Irradiated rats which did not receive the drug developed dense cataracts. Lenses from control rats which received no radiation remained transparent. Individual lenses were weighed, homogenized, and assayed for protein content using the Lowry method. The molecular weight distribution of soluble protein was determined by HPLC. Mean lens weights were: controls 48.2 mg; irradiated, drug-treated 45.9 mg; and irradiated, nontreated 45.5 mg. Protein accounted for over 40% of the lens weight in control and drug-treated rats and less than 20% for the nontreated cataractous lenses. Water was less than 60% of the lens weight in control and drug-treated rats and over 80% in cataractous lenses. Insoluble protein ranged from 12 to 17% of the total lens weight for each group. The ratio of insoluble to soluble lens protein was 0.40 for control, 0.65 for drug-treated, and 11.28 for cataractous rat lenses. HPLC confirmed a dramatic loss of soluble protein and a complete absence of protein below 25K daltons in cataractous lenses. Proteins below 25K daltons accounted for over 25% of the soluble protein in control and drug-treated rat lenses. WR-77913 stabilizes protein composition and appears to be an effective inhibitor of radiation cataractogenesis. PMID- 3025130 TI - Adenovirus type 3 infection with systemic manifestation in apparently normal children. AB - Between July 1983 and February 1984, eight children with adenovirus Type 3 infection, proven by virus isolation from sputum, stool or nasopharyngeal swabs and a fourfold increase in complement fixation antibody titers against the virus, were treated in our department. All eight patients had fever lasting at least 7 days, hepatomegaly, diffuse pulmonary infiltrates and abnormal liver function tests. Seven of the patients exhibited dyspnea and pulmonary wheezing. Six of the patients developed changes in state of consciousness, and three had repeated convulsions. EEG patterns in three of the patients were compatible with encephalopathy. Other clinical manifestations included: follicular tonsillitis in two patients, diarrhea in two, pneumothorax in one, and shock with disseminated intravascular coagulation in one. The spectrum of adenovirus Type 3 infection reported here has been described previously only in the viral hemorrhagic fevers. This adenovirus Type 3 infection shares the potential for disseminated disease that has been described previously for Type 7, simulating Reye's syndrome. PMID- 3025131 TI - Studies on the effect of intralipid on human monocyte functions in vitro. AB - Intralipid (IL) particles were ingested by human monocytes in culture. These particles remained within the cells for periods of up to 3 weeks in culture. The presence of IL particles did not alter the normal secretion of lysozyme or prostaglandin E2 (PGE2), nor was the normal process of biochemical activation altered, as evidenced by the expected increase in PGE2 secretion and superoxide anion production by stimulated monocytes. Phagocytosis of zymosan particles was increased in monocytes that had ingested IL. These results indicate that in this in vitro system essential monocyte functions were not altered following ingestion of IL. PMID- 3025132 TI - Sarcomatous degeneration of giant cell tumours. AB - Sarcomatous degeneration in giant cell tumours (G.C.T.) usually only occurs in patients of late adult age. Unlike other Centres, we have not up to now observed primary malignant G.C.T. In 327 cases registered at the Tumour Centre of the Rizzoli Institute, there were 10 cases of sarcomatous degeneration. In 8 of these, radiotherapy had been used as the initial treatment of the tumour. The incidence of sarcomatous degeneration when the radiation dose exceeded 4000r was 29%. The prognosis in such cases is very serious, the only feasible treatment being amputation. The possibility of sarcomatous degeneration must be considered before deciding to adopt radiotherapy as the method of primary treatment in giant cell tumours. PMID- 3025133 TI - [Extramammary Paget's disease associated with stomach cancer]. AB - A case of extramammary Paget's disease in the perianal region is reported. The disease presented as a whitish lesion with central erosion. Examination revealed an underlying adenocarcinoma of the stomach. The disease was treated by cryosurgery. PMID- 3025134 TI - Small cell carcinoma of the head and neck. AB - Treatment methods for patients with small cell carcinoma of the head and neck are changing. Increasing clinical experience with these tumors and a better appreciation of patterns of recurrence, the need for thorough tumor staging, and the importance of chemotherapy as a form of primary therapy have contributed to these changes. Current concepts regarding small cell carcinoma arising in the head and neck are reviewed. Reported results of treatment for a variety of head and neck sites and site-specific relapse rates are summarized. New cases of small cell carcinoma arising in the hypopharynx, paranasal sinus, and from an unknown primary are added to the reported experience in the literature. PMID- 3025135 TI - Metacarpal measurements in X-linked hypophosphataemic rickets. AB - In 15 girls and 7 boys with hypophosphataemic vitamin D-resistant rickets, midshaft diameter, combined cortical thickness, cortical area and metacarpal length were determined in the second metacarpal on radiographs of the left hand. The values obtained were compared with age-matched and height-matched controls. In a longitudinal study the same bone measurements were taken in these 22 patients and in additional 3 boys. The metacarpal diameter in the patients was increased whereas the combined cortical thickness was decreased. The former feature appeared to have its onset in early childhood. Cortical area was normal for age and increased for height, which was taken to indicate a normal or increased bone mass in these children. Metacarpal length was normal for age and height in the boys and increased for height but still within the normal range in girls. This is in agreement with the normal arm-span that has been found in these children. PMID- 3025136 TI - The Greig cephalopolysyndactyly syndrome. PMID- 3025137 TI - Effect of wheat bran and pectin on bile acid and cholesterol excretion in ileostomy patients. AB - The effect of addition of pectin or wheat bran to a constant low-fibre diet on bile acid and cholesterol excretion from the small intestine has been studied in ileostomy patients. The study was designed to minimize bacterial alteration of ileostomy contents. An addition of 15 g of citrus pectin increased bile acid excretion by 35 per cent (P less than 0.05) and net cholesterol excretion by 14 per cent (P less than 0.05) in six patients, while 16 g of wheat bran to another six patients caused no consistent change. Ileostomy fat excretion increased on the diet with added pectin (P less than 0.05) but not on that with bran. The results support the concept of dietary fibre exerting its effects on lipid metabolism by altering intestinal excretion of sterols. PMID- 3025138 TI - [Hearing loss caused by aminoglycoside antibiotics: affect on the membrane component PIP2 in outer hair cells as the mechanism of action]. AB - Experiments with live isolated outer hair cells are described that provide an explanation of the toxic mechanism of aminoglycoside antibiotics. Outer hair cells were isolated from the guinea pig cochlea by microsurgery and maintained in artificial perilymph. The presence of the membrane lipid PIP2 (phosphatidylinositol bisphosphate) and other components of the phosphoinositide cycle was demonstrated through their labeling with radioactive phosphate. The lipids play a key role in the function of outer hair cells as part of a second messenger and amplification system that controls the motility of these cells. Furthermore, it was shown that the ototoxic aminoglycoside gentamicin has a selective and extremely high binding affinity to PIP2. The specific toxicity of aminoglycoside antibiotics is explained by the inhibition of the function of PIP2. PMID- 3025139 TI - Resistance to suppression and acquisition of memory by human T cells. AB - Human memory cells acquire resistance to several types of suppressor cells, including MLR generated suppressor cells. These data suggest one possible mechanism of that resistance, namely retention of IL-2 receptors in the resting state. Cells from a 7-day MLR were separated on a single step percoll gradient. All proliferating cells were found in the interface. Pellet cells were nondividing. Interface and pellet cells had equivalent memory function in a secondary MLR. Thus, there appear to be at least two subpopulations of memory cells, including one that is primed without undergoing division. These subpopulations are functionally distinct. Interface memory cells were 30-50% more resistant than pellet memory cells to MLR generated suppressor cells. On culture day 10, neither pellet nor interface cells displayed significant spontaneous proliferation but exogenous Interleukin 2 (IL-2) produced up to five times as much proliferation in interface cells as in pellet cells. Further, FACS analysis with an anti-TAC equivalent antibody also showed that significantly more interface cells have surface receptors for IL-2. Thus, cells that had previously divided continue to have more and/or higher affinity receptors for IL-2 even after return to the resting state. If a mechanism of suppression in the mixed lymphocyte reaction is to reduce the synthesis/release of Il-2, memory cells may acquire their relative resistance to this suppression by virtue of the increased IL-2 sensitivity of this discrete subpopulation. PMID- 3025141 TI - Canary pox causing high mortality in an aviary. AB - In an aviary housing 200 six-month-old canaries, 165 became ill and 145 died over a 6-week period from a disease initially characterized by lethargy, ruffled feathers, open-mouth breathing, and death in 2 to 3 days. Proliferative "pox like" lesions around the eyes and mouth were not seen until the 4th week. At necropsy, initially affected birds had cloudy air sacs and patchy pneumonia. Histologically, the lungs had proliferative necrotizing bronchitis. Birds necropsied later had proliferative skin lesions and intracytoplasmic inclusions typical of poxvirus in the epidermis and airway epithelium. A virus was isolated from an organ pool of lung, air sac, liver, and skin of affected birds and was identified by electron microscopy as poxvirus. PMID- 3025140 TI - Chemodectoma of the carotid body and ganglion nodosum treated with radiation therapy. AB - A total of six chemodectomas of the carotid body or ganglion nodosum in four patients were treated with radiation therapy. The purpose of this paper is to present the results of treatment in these patients. One patient experienced complete regression of her bilateral carotid body tumors over the ensuing 2.5 years and the other 3 patients have had no evidence of disease progression of their chemodectomas for 2, 3, and 4.5 years, respectively. PMID- 3025142 TI - Effectiveness of canine parvovirus vaccines. PMID- 3025143 TI - Evaluation of a six-hour combined dexamethasone suppression/ACTH stimulation test in dogs with hyperadrenocorticism. AB - Seventeen dogs with hyperadrenocorticism were studied. Three dogs had functioning adrenocortical tumors and 14 had pituitary-dependent hyperadrenocorticism. Each dog was evaluated by determining the endogenous plasma ACTH concentration and by performing 4 tests: ACTH stimulation, dexamethasone screening, dexamethasone suppression, and a 6-hour combined dexamethasone suppression/ACTH stimulation test. The combined test was less reliable as a screening test in diagnosing hyperadrenocorticism than was the dexamethasone screening test or the ACTH stimulation test. Compared with the endogenous plasma ACTH concentration, results of the dexamethasone suppression portion of the combined test were less reliable in distinguishing dogs with adrenocortical tumors from those with pituitary dependent hyperadrenocorticism. It was concluded that the combined test cannot be recommended for use. PMID- 3025144 TI - Inhibition of biological actions of 12-O-tetradecanoylphorbol-13-acetate by inhibitors of protein kinase C. AB - Evidence has been accumulated that the phorbol ester receptor in intact cells is protein kinase C. However, it is not certain whether the various actions of 12-O tetradecanoylphorbol-13-acetate (TPA) on cultured cells are all mediated by the activation of protein kinase C. Therefore, we examined the effects of inhibitors of protein kinase C, palmitoylcarnitine (PC) and phloretin (PH), on several actions of TPA on cells. PC at the concentration of 30 micrograms/ml completely prevented the inhibitory actions of TPA on differentiation of Friend leukemic cells (FLC) induced by hexamethylene bisacetamide (HMBA) and on metabolic cooperation of V79 cells. PC, however, did not prevent the TPA-induced promotion of the transformation of BALB/3T3 cells, even at the concentration of 40 micrograms/ml. On the other hand, PH at the concentration of 30 micrograms/ml did not inhibit the actions of TPA to inhibit differentiation of FLC and metabolic cooperation of V79 cells, but completely inhibited the transformation of BALB/3T3 cells and its promotion by TPA. These results indicate that protein kinase C possibly mediates some of the actions of TPA, such as the inhibition of differentiation and metabolic cooperation, but not that of the promotion of in vitro transformation. PMID- 3025145 TI - Antitumor effect of adriamycin entrapped in liposomes conjugated with monoclonal antibody against tumor-associated antigen of bovine leukemia cells. AB - Monoclonal antibody against tumor-associated antigen (TAA) expressed on bovine leukemia cells was conjugated to liposomes containing adriamycin (ADM), and the specificity and therapeutic effects of the conjugates were examined in vitro and in vivo using a TAA-positive bovine leukemia cell line as the target tumor. In vitro studies with the TAA-positive cell line clearly indicated that the antibody conjugated liposomes containing ADM exerted selective effects on TAA-positive cells in the inhibition assay of 3H-thymidine incorporation. Three injections of liposomes containing ADM (4 mg/kg) into tumor-bearing nude mice significantly inhibited the tumor growth and the therapeutic effect of the antibody-conjugated liposomes was far greater than that of normal mouse IgG-conjugated liposomes as assessed in terms of tumor size. PMID- 3025146 TI - Direct evaluation of phenylacetyl-CoA: 6-aminopenicillanic acid acyltransferase of Penicillium chrysogenum by bioassay. AB - The enzyme phenylacetyl-CoA: 6-Aminopenicillanic acid acyltransferase of Penicillium chrysogenum was evaluated by direct bioassay against Micrococcus luteus ATCC 9341. The enzyme required dithiothreitol, was inactivated by 0.2 mM Hg2+ (100%), Zn2+ (80%), Cu2+ (60%), 1 mM N-ethylmaleimide (80%), and showed maximal catalytic activity at pH 8.4 and 20 degrees C. The V50 values for phenylacetyl-CoA and 6-aminopenicillanic acid were 0.55 mM and 1 microM, respectively. When octanoyl-CoA was employed as substrate similar results were obtained. In both cases the product generated showed strong antibacterial activity which was quickly lost when incubation was carried out with beta lactamase. Reactions performed in the presence of Escherichia coli penicillin acylase did not generated active products when phenylacetyl-CoA was the substrate; they did with octanoyl-CoA. Time-course experiments revealed that the highest enzyme levels are found in 36 hours mycelium and remained almost constant from 48 to 96 hours; thereafter the level of the enzyme slowly decreased. PMID- 3025147 TI - Oxetanocin, a novel nucleoside from bacteria. PMID- 3025148 TI - General aspects of virus drug resistance with special reference to herpes simplex virus. AB - The features of virus drug resistance are reviewed with examples from studies of herpes simplex virus drug-resistant mutants. Virus drug resistance, compared with drug resistance of bacteria or eukaryotes, is distinguished by its ability to provide information on drug selectivity. Identification of genes in which mutations arise to confer drug resistance defines gene products which contribute to antiviral selectivity. The gene products can then be dissected functionally with the aid of these mutations. Laboratory studies of the frequency of mutation to drug resistance and the features of drug-resistant mutants may have predictive value for the clinic. PMID- 3025150 TI - Molecular basis of drug resistance to new antirhinovirus agents. AB - The modes of action of antirhinovirus agents and mechanisms of resistance to the agents are compared. Chalcone Ro 09-0410, 4',6-dichloroflavan (DCF) and RMI 15,731, which were active against rhinoviruses, inactivated the virus directly. Inactivation was associated with the binding of the agents to the virus particles, since the infectivity, reduced by exposure to the compounds, was restored to the original level by extraction of the agents with chloroform. The binding of [3H]chalcone to rhinovirus type 2 was inhibited by any of the three unlabelled agents. Furthermore, virus sublines selected for resistance to both dichloroflavan and RMI-15,731 showed cross-resistance to chalcone and vice versa. These findings indicate that the three agents exert their activities through the same mode of action (namely binding to or interaction with a specific site on the viral capsid protein) and that the binding or interaction sites for these three agents are either the same or very close to each other. Since the sublines resistant to chalcone and to RMI-15,731 have little or no capability to bind to chalcone, the resistance to these agents is conferred by changes in the viral capsid protein. On the other hand, flavone Ro 09-0179 and enviroxime, which were active widely against picornaviruses, had no ability to inactivate the virus directly, and their antiviral activity was not associated with the capsid protein. PMID- 3025149 TI - Sensitivity monitoring of herpes simplex virus isolates from patients receiving acyclovir. AB - A simple plaque-reduction assay was used to determine the acyclovir sensitivity of herpes simplex virus isolates taken from patients enrolled int he acyclovir clinical trial programme. The resultant data revealed no reduction in acyclovir sensitivity in virus from those patients, with a normal immune status, receiving topical, oral or intravenous acyclovir for the treatment of acute disease episodes. In the treatment or prophylaxis of chronic herpes infections in immunocompromised patients reductions in sensitivity were observed but these were infrequent. Sensitive virus was later recovered from a small number of patients who had yielded resistant virus when they were followed through to the next recurrence. PMID- 3025151 TI - Susceptibility of HSV strains from patients with genital herpes treated with various formulations of acyclovir. AB - Virus isolates from 55 patients with recurrent or first-episode genital herpes, treated with various formulation of acyclovir, were tested for their susceptibility to the drug. In all but one of these patients pre-treatment isolates were available for comparison with later isolates. In none of patients, even those receiving long-term prophylaxis, was the susceptibility of the post treatment isolates significantly reduced. PMID- 3025152 TI - Clinical resistance to acyclovir of herpes simplex virus infections in immunocompromised patients. AB - Herpes simplex virus strains, isolated from three immunocompromised patients whose infections showed clinical resistance to acyclovir, were studied as treatment progressed. Virus isolated from two patients remained sensitive to acyclovir throughout. Isolates from the third patient, who had received a prolonged course of oral acyclovir, showed a sharp decrease in drug sensitivity which corresponded to loss of thymidine kinase activity. No changes in restriction endonuclease profiles were observed in isolates from the same patient as treatment with acyclovir progressed. PMID- 3025154 TI - The efficacy and tolerance of intranasal interferons: studies at the Common Cold Unit. AB - Intranasal sprays of interferons (IFNs) given one day before and for three days after virus challenge can protect human volunteers from infection with rhinoviruses, coronavirus, and influenza. Longer dosage of IFN gives rise to nasal symptoms and signs such as bloodstained nasal discharge. More effective IFNs and regimes are therefore needed. IFN beta is active but the degree to which it will irritate the nose is unknown. Combining IFNs with synthetic antiviral drugs can produce synergistic increases in antiviral activity. It is suggested that these increases may be exploited in future experiments. PMID- 3025153 TI - Characterization of acyclovir-resistant and -sensitive herpes simplex viruses isolated from a patient with an acquired immune deficiency. AB - Several different genital and non-genital HSV isolates were obtained from a patient with an acquired immune deficiency of unknown aetiology. The patient was initially treated with topical acyclovir (ACV) and later with topical and intravenous ACV. In spite of treatment with antiviral drugs the patient continued to shed virus and to have extensive genital ulcerations. Restriction endonuclease (RE) analyses of the viral DNA revealed that all the isolates had characteristic HSV-2 patterns and that there were three genetically distinct virus groups among the ten isolates tested. Three post-therapy isolates, with the same RE pattern, were found to be devoid of thymidine kinase activity (TKD), highly resistant to ACV in cell culture, but sensitive to vidarabine (ara-A), phosphonoacetate and phosphonoformate. Two of these TKD isolates were obtained during and after topical ACV therapy and before intravenous treatment. Mice inoculated intracerebrally with a lethal dose of each of the three TKD viruses were refractory to ACV, but responded to vidarabine or a combination of ACV and ara-A. Mice inoculated with the TK+ viruses (including the pre-therapy isolate) responded to ACV and/or ara-A treatment. The results indicate that: (i) TKD variants may be produced in humans after topical ACV therapy; (ii) different ACV resistant or sensitive HSV-2 variants can establish latency at different body sites and reactivate; and (iii) when drug-resistant viruses are isolated from patients with multiple reactivations, the drug in question should not be discontinued, since the patients may also be shedding drug-sensitive virus at a different body site. PMID- 3025155 TI - Anti-herpes virus combinations in relation to drug resistance. AB - Two broad categories of combinations have been subject to experimental investigation against HSV-1 and HSV-2; those consisting of an active antiviral drug and an inhibitor of its catabolism and those which contain two active antiviral agents. The latter seek to exploit biochemical mechanisms likely to lead to a synergistic or complementary interaction. Examples of both types of combination for the suppression of resistance development and for the treatment of infection caused by resistant strains are discussed. Their possible relevance to the latent state is considered. PMID- 3025156 TI - The antiviral activity against herpes simplex virus of the triterpenoid compounds carbenoxolone sodium and cicloxolone sodium. AB - Dose-response experiments show that the presence of 300 microM cicloxolone sodium (CCX) or 500 microM carbenoxolone sodium (CBX) during the HSV replication cycle reduced the infectious virus yield by 10,000- to 100,000-fold: CCX is the more potent anti-herpes agent. HSV-2 replication was consistently more severely restricted by either drug than was that of HSV-1. The ED50 values obtained for either drug against HSV-1 or HSV-2 correlate well with data from dose-response curves. CCX, and to a lesser extent CBX, can be cytotoxic but the degree of cytotoxicity varies between cell lines and is also affected by the physiological state of the cells. Triterpenoid drugs exhibit some activity against virus particles in suspension but the effect is small and contributes little to the overall antiviral effect. The drugs appear to be active throughout the replication cycle. In contrast to all other anti-herpesvirus agents in clinical use the triterpenoid compounds do not appear to act directly to block virus DNA synthesis. HSV mutants resistant to ACG and PAA, or lacking a thymidine kinase gene, appear as sensitive as wild-type virus to CCX inhibition. HSV growth in the presence of the drugs resulted in a lower number of assembled virus particles but reduced to a much greater extent the infectious virus yield: thus the progeny virus quality is greatly diminished. Thermolability of progeny virus correlated well with this diminution of quality in increasing CCX concentration. SDS PAGE analysis of the structural proteins of virus particles made in cells treated with 300 microM CCX revealed numerous differences in the relative intensities of protein bands, which is in keeping with the changed quality of the drug-produced virus. SDS PAGE analysis of the polypeptides induced in drug treated infected cells revealed two effects; some polypeptides were synthesised in reduced amounts and the nuclear/cytoplasmic distribution of certain proteins was affected. Post translational processing by glycosylation and sulphation of both cellular and HSV induced proteins was strongly inhibited by the triterpenoid drugs, while phosphorylation of only a few polypeptides appeared to be affected. PMID- 3025157 TI - Clinical and laboratory experience with acyclovir-resistant herpes viruses. AB - The emerging field of antiviral sensitivity testing must depend upon some of the lessons learned from prior experience with anti-bacterial sensitivity testing, but also take into account additional factors including intracellular drug distribution and metabolism as well as the unique features of viral replication. There is an urgent need to standardise in-vitro sensitivity testing, and an international collaborative study is essential so that different laboratories may compare and define their sensitivity results in the context of the outcome of therapy in infected patients. It is only by such a study that the influence of patient and therapeutic factors such as immune function, anatomical location of the infection and dosage regimens used can be understood. PMID- 3025159 TI - Induction of acyclovir-resistant mutants of herpes simplex virus type I in athymic nude mice. AB - Induction of acyclovir resistance was studied in the immune compromised host by multiple passage of plaque-purified herpes simplex type I strains in athymic nude mice receiving suboptimal antiviral therapy. Mice infected with a highly pathogenic clinical isolate rapidly developed infections that were resistant to therapy. Viruses isolated from these mice had decreased in-vitro sensitivities to acyclovir, as well as altered characteristics when assayed by [125I]plaque autoradiography. In contrast, less virulent laboratory strains, or a genetically stable clinical isolate, showed no indication of mutation to resistance after extended passage in this mouse model. Highly pathogenic viruses may increase the probability of mutation to resistance because of the large amount of infectious virus they produce, while viruses of equivalent virulence may produce different amounts of drug-resistant progeny because of alterations in the replication fidelity of the viral DNA polymerase. PMID- 3025158 TI - Extended acyclovir therapy for herpes genitalis: changes in virus sensitivity and strain variation. AB - The in-vitro sensitivity of herpes simplex virus isolated from patients enrolled in clinical trials of orally administered chronic suppressive acyclovir therapy was determined. Approximately 500 isolates from 250 patients were examined. There was no increased incidence of recovery of virus less sensitive to acyclovir in vitro following chronic administration of acyclovir or intermittent therapy for recurrences during treatment periods of four months to four years. Changes in in vitro sensitivity of virus isolated from two patients who shed virus less sensitive to acyclovir were studied to determine the possible role of continued antiviral therapy in selecting for progressively more resistant phenotypes. Increasing resistance was not documented. Both patients who shed less sensitive virus derived clinical benefit from chronic orally administered acyclovir treatment. PMID- 3025160 TI - Cyclic AMP, prostaglandins and leukaemia: relevance to lithium-induced leukaemia. PMID- 3025162 TI - Non-starch polysaccharides as a protective factor in human large bowel cancer. AB - The hypothesis that lack of dietary fibre in the diet is responsible for a variety of large bowel problems, including cancer, has stimulated much discussion and research over the past 15 years. However, the epidemiological examination of this hypothesis has been hampered by the absence of data on the fibre content of most of the world's foods. In Scandinavia and Britain where the consumption of the major chemical fraction of dietary fibre, the non-starch polysaccharides has been measured using accurate methods, significant negative associations have been shown with large bowel cancer occurrence. These studies suggest that non-starch polysaccharides may be protective in populations at otherwise high risk of large bowel cancer from an excess of meat and fat. However, methodological problems in the assessment of non-starch polysaccharide consumption in individuals preclude the use of case control studies in verifying these associations within a single homogeneous population. PMID- 3025161 TI - Significance of L-ascorbic acid and urinary electrolytes in promotion of rat bladder carcinogenesis. AB - The present studies report on the significance of L-ascorbic acid (AA) and urinary electrolytes for promotion of rat urinary bladder carcinogenesis. Male F344 rats were given an oral administration of 0.05% N-butyl-N-(4 hydroxybutyl)nitrosamine (BBN) as an initiator for 4 weeks, and were then subjected to treatment with dietary supplements of test chemicals for 32 weeks. Administration of 5% sodium L-ascorbate (SA), the sodium ion form of AA significantly promoted urinary bladder carcinogenesis, whereas administration of 5% AA did not. The urine of rats given SA but not AA was characterized by an apparent elevation of pH, an increase of sodium ion concentration, and increases in the urinary content of total AA and its metabolite, dehydroascorbic acid. Administration of 3% NaHCO3, which induced elevation of pH and increase of sodium ion concentration in the urine, promoted BBN bladder carcinogenesis. When rats were given 5% AA plus 3% NaHCO3, AA enhanced the promoting activity of NaHCO3. Lowering of pH by 1% NH4Cl clearly reduced the promoting activity of 5% SA when these two compounds were given concurrently. Treatment with 5% AA plus 3% K2CO3 promoted BBN bladder carcinogenesis in rats, whereas addition of 5% CaCO3 or 5% MgCO3 to AA did not. These results strongly indicate the important role of urinary sodium or potassium ion concentration and pH in modulating urinary bladder carcinogenesis by AA. PMID- 3025163 TI - Measurement of individual aflatoxin exposure among people having different risk to primary hepatocellular carcinoma. AB - The establishment of the carcinogenic role of aflatoxins has been impeded by the lack of suitable tests to measure individual exposure for long-term studies. It was shown that the use of immuno-concentration followed by high pressure liquid chromatography (HPLC) or immunoassay can regularly detect aflatoxins down to pg/ml in fluids including urine. Aflatoxin M1 (AFM1) in free form was identified as the major metabolite in urine suitable for use as an approximate dosimetric indicator of recent exposure to aflatoxin B1 (AFB1). A panel of monoclonal antibodies of IgG class were developed in the murine system against AFB1 and/or AFM1. Their affinity constants are at the level of 10(8)-10(9) L/mol, and they are suitable for use in either effective immuno-concentration or in high sensitivity immunoassays. It was shown in a high risk area of liver cancer (Qidong of China) that 10% or more of the local inhabitants had a urinary output of AFM1 one or two orders of magnitude higher than that of Beijing people, especially during the wet seasons. The ingestion of AFB1 among these local people was estimated to exceed 1 mg/year. The major source came from contaminated corn and rice, but local alcoholic beverages also contributed. The increased excretion of AFM1 was more pronounced in patients with chronic active hepatitis. This observation of very low AFM1 content in urine of a significant percentage of local people implies their food storage practices may offer a feasible method for such prevention. The value and possible problems in conducting long-term studies on primary prevention are discussed. PMID- 3025164 TI - Dietary fibre in the Japanese diet. AB - The low risk of colon cancer among the Japanese suggests their high intake of dietary fibre. Composite diet mixtures for 1959, 1970, and 1979 were prepared utilising the food consumption data from the National Nutrition Survey in Japan and analysed for non-starch polysaccharides (NSP) at the Dunn Clinical Nutrition Centre in Cambridge. The results showed that average intake of NSP by a Japanese in the above years did not exceed 13 g per day, which is as low as the corresponding intake by the Scandinavians and the British whose risk of colon cancer is known to be high. Since all the materials were analysed by the same methods and at the same laboratory, the results are well comparable and indicate that the low risk of colon cancer among the Japanese cannot readily be explained by their intake of NSP alone. PMID- 3025165 TI - Diet and nutrition as risk factors for cancer. AB - Involvement of diet and nutrition as risk factors for human cancers has been established by two general types of evidence. Epidemiological studies in human populations have identified associations between patterns of incidence for various forms of cancer and diet composition or food consumption patterns. Such associations are particularly evident in studies on migrant populations and in patterns of geographic localization of specific forms of cancer. Reduction in gastric cancer (and increase in colon cancer) incidence has been observed in immigrants from Japan to the U.S. via Hawaii with concurrent change in dietary habits. High fat consumption has been linked to increased incidence of breast and colon cancer, and evidence is accumulating that suggests increased intake of dietary fiber (or some specific component of it) may be associated with diminished risk of colorectal cancer. Laboratory studies have demonstrated the existence of dietary constituents that might impact cancer risk in people consuming them. Substances in this class fall into two general categories: genotoxic carcinogens/mutagens; and protective factors which inhibit experimentally-induced chemical carcinogenesis in animals. Numerous naturally occurring carcinogens/mutagens have been identified. Examples include aflatoxins, cycasin, and bracken fern carcinogen(s), among others. Genotoxic substances associated with the use of intentional food additives are the N-nitroso compounds formed as nitrosation products from nitrite. Potential carcinogenic risk from food constituents also comes from mutagens found in foods as natural components, or formed in the course of cooking.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3025166 TI - Cancer, diet, and public policy. AB - The response of Government to the relationship between diet and chronic disease is a legitimate public health policy issue, yet the proper role of Government is a subject of continuing debate. Regulatory mechanisms designed to address questions of food additives or contaminants in food are ill-suited to address questions of diet/disease relationships because of the different standards and types of evidence involved. In this paper we examine, through the use of several examples, the question of whether current evidence on the relationship of diet to cancer is sufficient to make changes in public health policy. Finally, we discuss potential Governmental approaches, both short- and long-term, to implementing policy in the area of diet and cancer. PMID- 3025167 TI - Characterization of the ftsB gene as an allele of the nrdB gene in Escherichia coli. AB - A temperature-sensitive, salt-rescuable ftsB cell division mutant, MFT84, was found to be hydroxyurea sensitive on low-salt medium. Complementation studies with plasmids and a marker rescue study with bacteriophage M13 nrd indicated that ftsB is an allele of nrdB and that the mutation occurs in the region corresponding to nucleotides 6729 to 7032 of the nrdB gene. Enzymatic characterization demonstrated that the B2 subunit of ribonucleoside-diphosphate reductase encoded by ftsB was responsible for the decreased activity and the thermolability of the enzyme. The ftsB-encoded B2 subunit was activated by the addition of 0.1 M NaCl to an in vitro assay, corroborating the in vivo temperature-dependent salt requirement was a result of a defective B2 subunit. PMID- 3025169 TI - Negative modulation of Escherichia coli NAD kinase by NADPH and NADH. AB - NAD kinase was purified 93-fold from Escherichia coli. The enzyme was found to have a pH optimum of 7.2 and an apparent Km for NAD+, ATP, and Mg2+ of 1.9, 2.1, and 4.1 mM, respectively. Several compounds including quinolinic acid, nicotinic acid, nicotinamide, nicotinamide mononucleotide, AMP, ADP, and NADP+ did not affect NAD kinase activity. The enzyme was not affected by changes in the adenylate energy charge. In contrast, both NADH and NADPH were potent negative modulators of the enzyme, since their presence at micromolar concentrations resulted in a pronounced sigmoidal NAD+ saturation curve. In addition, the presence of a range of concentrations of the reduced nucleotides resulted in an increase of the Hill slope (nH) to 1.7 to 2.0 with NADH and to 1.8 to 2.1 with NADPH, suggesting that NAD kinase is an allosteric enzyme. These results indicate that NAD kinase activity is regulated by the availability of ATP, NAD+, and Mg2+ and, more significantly, by changes in the NADP+/NADPH and NAD+/NADH ratios. Thus, NAD kinase probably plays a role in the regulation of NADP turnover and pool size in E. coli. PMID- 3025168 TI - Isolation of the outer membrane and characterization of the major outer membrane protein from Spirochaeta aurantia. AB - The outer membrane of Spirochaeta aurantia was isolated after cells were extracted with sodium lauryl sarcosinate and was subsequently purified by differential centrifugation and KBr isopycnic gradient centrifugation. The purified outer membrane was obtained in the form of carotenoid-containing vesicles. Four protein species with apparent molecular weights of 26,000 (26K), 36.5K, 41K, and 48.5K were readily observed as components of the vesicles. The 36.5K protein was the major polypeptide and constituted approximately 90% of the outer membrane protein observed on sodium dodecyl sulfate-polyacrylamide gels. Under mild denaturing conditions the 36.5K major protein exhibited an apparent molecular weight of approximately 90,000. This, together with the results of protein cross-linking studies, indicates that the 36.5K polypeptide has an oligomeric conformation in the native state. Reconstitution of solubilized S. aurantia outer membrane into lipid bilayer membranes revealed the presence of a porin, presumably the 36.5K protein, with an estimated channel diameter of 2.3 nm based on the measured single channel conductance of 7.7 nS in 1 M KCl. PMID- 3025170 TI - Reconstitution of pyrroloquinoline quinone-dependent D-glucose oxidase respiratory chain of Escherichia coli with cytochrome o oxidase. AB - D-Glucose dehydrogenase is a pyrroloquinoline quinone-dependent primary dehydrogenase linked to the respiratory chain of a wide variety of bacteria. The enzyme exists in the membranes of Escherichia coli, mainly as an apoenzyme which can be activated by the addition of pyrroloquinoline quinone and magnesium. Thus, membrane vesicles of E. coli can oxidize D-glucose to gluconate and generate an electrochemical proton gradient in the presence of pyrroloquinoline quinone. The D-glucose oxidase-respiratory chain was reconstituted into proteoliposomes, which consisted of two proteins purified from E. coli membranes, D-glucose dehydrogenase and cytochrome o oxidase, and E. coli phospholipids containing ubiquinone 8. The electron transfer rate during D-glucose oxidation and the membrane potential generation in the reconstituted proteoliposomes were almost the same as those observed in the membrane vesicles when pyrroloquinoline quinone was added. The results demonstrate that the quinoprotein, D-glucose dehydrogenase, can reduce ubiquinone 8 directly within phospholipid bilayer and that the D-glucose oxidase system of E. coli has a relatively simple respiratory chain consisting of primary dehydrogenase, ubiquinone 8, and a terminal oxidase. PMID- 3025171 TI - Insertion-sequence-dependent rearrangements of Pseudomonas cepacia plasmid pTGL1. AB - Pseudomonas cepacia 249 (ATCC 17616) harbors a 170-kilobase (kb) plasmid designated pTGL1. We identified three insertion sequences, IS405, IS408, and IS411, on this plasmid. Various prototrophic and auxotrophic derivatives in our collection contained variants of pTGL1 formed by accretion and deletion of other elements. Plasmid pTGL6, the variant in one prototroph, evolved from pTGL1 by the addition of three copies of IS401 (1.3 kb) and one of IS402 (1 kb), to generate pTGL5, and recombination between two of the copies of IS401 on pTGL5 to form pTGL6. The latter event entailed loss of one copy of IS401 and an additional 5.4 kb of plasmid DNA. Derivatives of the broad-host-range plasmid pRP1 carrying the above insertion sequences and recombinant plasmids carrying fragments of plasmids pTGL6 and pTGL5 were used as probes to ascertain the extent of reiteration of the various elements in the P. cepacia genome. The data indicate a high frequency of genomic rearrangements which presumably contributes to the extraordinary adaptability of this bacterium. PMID- 3025173 TI - Biosynthesis and degradation of nodule-specific Rhizobium loti compounds in Lotus nodules. AB - Two nodule-specific Rhizobium loti compounds were identified in Lotus tenuis and Lotus pedunculatus nodules induced by strain NZP2037. One, a silver nitrate positive cation called rhizolotine, has been characterized as the riboside of a novel alpha-hydroxyimino acid containing a 1,4,5,6-tetrahydropyrimidine ring (G. J. Shaw, R. D. Wilson, G. A. Lane, L. D. Kennedy, D. B. Scott, and G. J. Gainsford, J. Chem. Soc. Chem. Commun., p. 180-181, 1986), and the other, yellow 1, stains yellow with ninhydrin. Both compounds were degraded by R. loti NZP2037 but not by strains of Rhizobium meliloti, Rhizobium trifolii, or Agrobacterium tumefaciens. Under the conditions tested neither compound was able to serve as a sole source of C or N for growth of R. loti NZP2037. Rhizolotine and yellow-1 were found in nodules from a range of different legumes inoculated with NZP2037, suggesting that the Rhizobium and not the host plant determines their synthesis. Neither compound was found in nodulelike structures of L. pedunculatus induced by transposon Tn5-induced noninfectious (Inf-) mutants of NZP2037 or in similar structures induced by a transconjugant of NZP2037 containing the symbiotic (Sym) cointegrate plasmid pPN1 of R. trifolii. Both compounds were also absent in the ineffective nodules induced by the bacterial-release-negative (Bar-) mutant, strain PN239. However, both compounds were present in nodules induced by the fixation-negative (Fix-) mutant PN235 and in Fix+ nodules formed by a plasmid cured derivative of NZP2037. These results would suggest that infection and bacterial release from the infection thread are necessary for nodule (symbiotic) synthesis of these compounds. PMID- 3025172 TI - Complementation of nitrogen-regulatory (ntr-like) mutations in Rhodobacter capsulatus by an Escherichia coli gene: cloning and sequencing of the gene and characterization of the gene product. AB - In vivo genetic engineering by R' plasmid formation was used to isolate an Escherichia coli gene that restored the Ntr+ phenotype to Ntr- mutants of the photosynthetic bacterium Rhodobacter capsulatus (formerly Rhodopseudomonas capsulata; J. F. Imhoff, H. G. Truper, and N. Pfenning, Int. J. Syst. Bacteriol. 34:340-343, 1984). Nucleotide sequencing of the gene revealed no homology to the ntr genes of Klebsiella pneumoniae. Furthermore, hybridization experiments between the cloned gene and different F' plasmids indicated that the gene is located between 34 and 39 min on the E. coli genetic map and is therefore unlinked to the known ntr genes. The molecular weight of the gene product, deduced from the nucleotide sequence, was 30,563. After the gene was cloned in an expression vector, the gene product was purified. It was shown to have a pI of 5.8 and to behave as a dimer during gel filtration and on sucrose density gradients. Antibodies raised against the purified protein revealed the presence of this protein in R. capsulatus strains containing the E. coli gene, but not in other strains. Moreover, elimination of the plasmid carrying the E. coli gene from complemented strains resulted in the loss of the Ntr+ phenotype. Complementation of the R. capsulatus mutations by the E. coli gene therefore occurs in trans and results from the synthesis of a functional gene product. PMID- 3025174 TI - Escherichia coli dnaK null mutants are inviable at high temperature. AB - DnaK, a major Escherichia coli heat shock protein, is homologous to major heat shock proteins (Hsp70s) of Drosophila melanogaster and humans. Null mutations of the dnaK gene, both insertions and a deletion, were constructed in vitro and substituted for dnaK+ in the E. coli genome by homologous recombination in a recB recC sbcB strain. Cells carrying these dnaK null mutations grew slowly at low temperatures (30 and 37 degrees C) and could not form colonies at a high temperature (42 degrees C); furthermore, they also formed long filaments at 42 degrees C. The shift of the mutants to a high temperature evidently resulted in a loss of cell viability rather than simply an inhibition of growth since cells that had been incubated at 42 degrees C for 2 h were no longer capable of forming colonies at 30 degrees C. The introduction of a plasmid carrying the dnaK+ gene into these mutants restored normal cell growth and cell division at 42 degrees C. These null mutants showed a high basal level of synthesis of heat shock proteins except for DnaK, which was completely absent. In addition, the synthesis of heat shock proteins after induction in these dnaK null mutants was prolonged compared with that in a dnaK+ strain. The well-characterized dnaK756 mutation causes similar phenotypes, suggesting that they are caused by a loss rather than an alteration of DnaK function. The filamentation observed when dnaK mutations were incubated at a high temperature was not suppressed by sulA or sulB mutations, which suppress SOS-induced filamentation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3025175 TI - DNA element of Corynebacterium diphtheriae with properties of an insertion sequence and usefulness for epidemiological studies. AB - The segment of DNA which is inserted within the tox gene of bacteriophage gamma and is responsible for its Tox- phenotype was found to be present and repeated approximately 30 times in the chromosome of Corynebacterium diphtheriae Belfanti 1030. Other C. diphtheriae strains contained a variable number of copies (1 to 25) of the same element. Sequence analysis showed that this repeated and interspersed DNA element was flanked by 9-base-pair direct repeats and that the 5' and 3' ends of the insertion contained sequences forming an imperfect inverted repeat. Therefore, the DNA segment here described has most of the typical structural features of a bacterial insertion sequence element. We show that different C. diphtheriae isolates derived from the same outbreak of diphtheria have an identical genomic distribution of this DNA element and that such DNA can be useful for epidemiological studies. PMID- 3025176 TI - Characterization of nonattaching mutants of Agrobacterium tumefaciens. AB - The first step in tumor formation by Agrobacterium tumefaciens is the site specific binding of the bacteria to plant host cells. Transposon mutants of the bacteria which fail to attach to carrot suspension culture cells were isolated. These mutants showed no significant attachment to carrot cells with either microscopic or viable cell count assays of bacterial binding. The nonattaching mutants were all avirulent. When revertants of the mutants were obtained by enriching for bacteria which do bind to carrot cells, the bacteria were found to have regained the ability to bind to carrot cells and virulence simultaneously. These results suggest that the ability of the bacteria to bind to plant cells is required for virulence. Like the parent strain, all of the nonattaching mutants synthesized cellulose, but unlike the parent strain, they failed to aggregate carrot suspension culture cells. The transposon Tn5, which was used to obtain the mutants, was located on a 12-kilobase EcoRI fragment of the bacterial chromosomal DNA in all of the nonattaching mutants from strain C58. That the mutant phenotype was due to the Tn5 insertion was shown by cloning the Tn5-containing DNA fragment from the mutant bacteria and using it to replace the wild-type fragment in the parent strain by marker exchange. The resulting bacteria had the same mutant phenotype as the original Tn5 mutants; they did not attach to carrot cells, they did not cause the aggregation of carrot cells, and they were avirulent. No difference was seen between the parent strain and the nonattaching mutants in hydrophobicity, motility, flagella, fimbriae, beta-2-glucan content, size of lipopolysaccharide, or ability of the lipopolysaccharide to inhibit bacterial attachment to tissue culture cells. Differences were seen between the parent strain and the nonattaching mutants in the polypeptides removed from the bacteria during the preparation of spheroplasts. Three of the mutants were lacking a polypeptide of about 34 kilodaltons (kDa). One mutant was lacking the 34-kDa polypeptide and another polypeptide of about 38 kDa. The fifth mutant was lacking a polypeptide slightly smaller than the 34-kDa polypeptide missing in the other four mutants. These missing polypeptides all reappeared in the revertants of the mutants. Thus, bacterial binding to plant cells appears to require the presence of these polypeptides. PMID- 3025177 TI - Expression of the gene for NAD-dependent glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides cloned in Escherichia coli K-12. AB - The structural gene (zwf) for Leuconostoc mesenteroides glucose-6-phosphate dehydrogenase has been cloned into Escherichia coli on pBR322 by selecting for complementation of a zwf mutation of E. coli which eliminates glucose-6-phosphate dehydrogenase. The gene has been subcloned on a 3.4-kilobase DNA segment. The encoded glucose-6-phosphate dehydrogenase displays the dual nucleotide specificity (reacting with NAD and NADP) of the L. mesenteroides enzyme and has a subunit Mr of 55,000. A second gene encoding protein of subunit Mr 24,000 is also encoded on the 3.4-kilobase DNA segment. The genes have been located by the analysis of deletions and insertions generated in vitro and by transcriptional mapping with a promoterless chloramphenicol acetyl transferase cartridge inserted at different sites in the 3.4-kilobase fragment. PMID- 3025178 TI - Cloning and analysis of ermG, a new macrolide-lincosamide-streptogramin B resistance element from Bacillus sphaericus. AB - To analyze the regulation of a newly discovered macrolide-lincosamide streptogramin B resistance element (ermG) found in a soil isolate of Bacillus sphaericus, we cloned this determinant and obtained its DNA sequence. Minicell analysis revealed that ermG specifies a 29,000-dalton protein, the synthesis of which is induced by erythromycin. S1 nuclease mapping was used to identify the transcriptional start site. These experiments demonstrated the presence on the ermG mRNA of a 197 to 198-base leader. Within the leader are two small open reading frames (ORFs) capable of encoding 11- and 19-amino-acid peptides. Each ORF is preceded by a suitably spaced Shine-Dalgarno sequence. The ermG protein is encoded by a large ORF that encodes a 244-amino-acid protein, in agreement with the minicell results. This protein and the 19-amino-acid peptide are highly homologous to the equivalent products of ermC and ermA. We conclude, on the basis of this homology, that ermG encodes an rRNA transmethylase. The leader of ermG can be folded into a structure that sequesters the Shine-Dalgarno sequence and start codon for the large ORF (SD3). On the basis of these data and on the observed greater responsiveness of the ermG system than of the ermC system to low concentrations of erythromycin, we propose a model for the regulation of this gene in which the stalling of a ribosome under the influence of an inducer, while reading either peptide, suffices to uncover SD3 and allow translation of the rRNA transmethylase. The evolution of ermG is discussed. PMID- 3025179 TI - Gene algD coding for GDPmannose dehydrogenase is transcriptionally activated in mucoid Pseudomonas aeruginosa. AB - Transcriptional regulation of alginate biosynthesis by Pseudomonas aeruginosa was studied. A DNA region complementing the alg-5 mutation within the alginate gene cluster was found by RNA-DNA dot blot and Northern hybridization to be transcriptionally activated in mucoid P. aeruginosa. This region was subcloned as a 3.2-kilobase BglII-ClaI DNA fragment on the broad-host-range controlled transcription vector pMMB24, and gene products were analyzed by expression from the tac promoter. A 48-kilodalton polypeptide was detected in extracts of P. aeruginosa and 35S-labeled Escherichia coli maxicells. By using the same expression system, GDPmannose dehydrogenase activity was detected in both P. aeruginosa and E. coli. Thus, gene algD coding for this enzyme was found to be present in the transcriptionally active DNA area. Insertion of the xylE gene within the BglII-ClaI fragment disrupted the induction of the 48-kilodalton polypeptide, GDPmannose dehydrogenase activity, and alg-5 complementing ability. With the algD-xylE transcription fusion, activation of algD gene expression was shown to occur in mucoid P. aeruginosa of different origins. In addition, regulation of the algD promoter activity was demonstrated to be mediated by a diffusible factor. PMID- 3025180 TI - Phenol: a complex chemoeffector in bacterial chemotaxis. AB - Earlier observations that phenol is a repellent for Salmonella typhimurium but an attractant for Escherichia coli were confirmed. This behavioral difference was found to correlate with a difference in the effect phenol had on receptor methylation levels; it caused net demethylation in S. typhimurium but net methylation in E. coli. On the basis of mutant behavior and measurement of phenol stimulated methylation, the attractant response of E. coli was shown to be mediated principally by the Tar receptor. In S. typhimurium, two receptors were found to be sensitive to phenol, namely, an unidentified receptor, which mediated the repellent response and showed phenol-stimulated demethylation; and the Tar receptor, which (as with E. coli) mediated the attractant response and showed phenol-stimulated methylation. In wild-type S. typhimurium, the former receptor dominated the Tar receptor, with respect to both behavior and methylation changes. However, when the amount of Tar receptor was artificially increased by the use of Tar-encoding plasmids, S. typhimurium cells exhibited an attractant response to phenol. No protein analogous to the phenol-specific repellent receptor was evident in E. coli, explaining the different behavioral responses of the two species toward phenol. PMID- 3025181 TI - Characterization of the specific pyruvate transport system in Escherichia coli K 12. AB - A mutant of Escherichia coli K-12 lacking pyruvate dehydrogenase and phosphoenolpyruvate synthase was used to study the transport of pyruvate by whole cells. Uptake of pyruvate was maximal in mid-log phase cells, with a Michaelis constant for transport of 20 microM. Pretreatment of the cells with respiratory chain poisons or uncouplers, except for arsenate, inhibited transport up to 95%. Lactate and alanine were competitive inhibitors, but at nonphysiological concentrations. The synthetic analogs 3-bromopyruvate and pyruvic acid methyl ester inhibited competitively. The uptake of pyruvate was also characterized in membrane vesicles from wild-type E. coli K-12. Transport required an artificial electron donor system, phenazine methosulfate and sodium ascorbate. Pyruvate was concentrated in vesicles 7- to 10-fold over the external concentration, with a Michaelis constant of 15 microM. Energy poisons, except arsenate, inhibited the transport of pyruvate. Synthetic analogs such as 3-bromopyruvate were competitive inhibitors of transport. Lactate initially appeared to be a competitive inhibitor of pyruvate transport in vesicles, but this was a result of oxidation of lactate to pyruvate. The results indicate that uptake of pyruvate in E. coli is via a specific active transport system. PMID- 3025182 TI - Transcription control of the aroP gene in Escherichia coli K-12: analysis of operator mutants. AB - The nucleotide sequence of the region containing the promoter-operator for the aroP gene was determined. The start site of aroP transcription was identified by using S1 nuclease mapping and primer extension techniques. Examination of the nucleotide sequence revealed the presence of two "TYR R" boxes which are similar to those identified in the regulatory regions of other genes in the tyrR regulon. Bisulfite-induced aroP operator-constitutive mutants were analyzed, and the base pair changes responsible for alterations in aroP regulation were located within these boxes. PMID- 3025183 TI - Molecular cloning and expression of the 3-chlorobenzoate-degrading genes from Pseudomonas sp. strain B13. AB - The genes specifying the utilization of 3-chlorobenzoate by Pseudomonas sp. strain B13 WR1 have been cloned by using a broad-host-range cosmid cloning system. Analysis of the catabolic products of the enzymatic reactions encoded by two hybrid cosmids, pMW65 and pMW90, by thin-layer and high-performance liquid chromatography demonstrated that both encoded the genes for the complete catabolism of 3-chlorobenzoate. Physical analysis of one of the cosmid derivatives, pMW65, by restriction endonuclease mapping and subcloning demonstrated that the pathway genes are encoded on a fragment no larger than 11 kilobases. PMID- 3025184 TI - Isolation of competition-defective mutants of Rhizobium fredii. AB - We coupled Tn5 mutagenesis with a competition assay to isolate mutants of Rhizobium fredii USDA 257 that are defective in competition for nodulation of soybeans. Two mutants with single Tn5 inserts in the chromosome showed reduced competitiveness in vermiculite but were identical to the wild-type strain in symbiotic properties when inoculated alone. Recombination of Tn5 and flanking genomic regions cloned from the mutants into the parent strain showed that Tn5 was responsible for the mutant phenotype. PMID- 3025186 TI - Transposition of IS91 does not generate a target duplication. AB - We determined the DNA sequences surrounding the junctions of IS91 in two insertion derivatives: pSU234 (pACYC184::IS91) and pSU240 (pBR322::IS91). The termini of IS91 consist of two imperfect inverted repeats eight base pairs long. Their sequence is 5'-TCGAGTAGG. . . CCTATCGA-3'. Insertion of IS91 did not generate direct repetitions in the target DNAs. PMID- 3025185 TI - Genetic and molecular characterization of the genes involved in short-chain fatty acid degradation in Escherichia coli: the ato system. AB - The structural organization and regulation of the genes involved in short-chain fatty acid degradation in Escherichia coli, referred to as the ato system, have been studied by a combination of classic genetic and recombinant DNA techniques. A plasmid containing a 6.2-kilobase region of the E. coli chromosome was able to complement mutations in the ato structural genes, atoA (acetyl-coenzyme A [CoA]:acetoacetyl [AA]-CoA transferase) and atoB (thiolase II), as well as mutations in the ato regulatory locus, atoC. Complementation studies performed with mutants defective in acetyl-CoA:AA-CoA transferase suggest that two loci, atoD and atoA, are required for the expression of functional AA-CoA transferase. The ato gene products were identified by in vitro transcription and translation and maxicell analysis as proteins of 48, 26.5, 26, and 42 kilodaltons for atoC, atoD, atoA, and atoB, respectively. In vitro and insertional mutagenesis of the ato hybrid plasmid indicated that the ato structural genes were arranged as an operon, with the order of transcription atoD-atoA-atoB. Although transcribed in the same direction as the atoDAB operon, the atoC gene appeared to use a promoter which was distinct from that used by the atoDAB operon. A delta atoC plasmid expressed the atoD, atoA, and atoB gene products only in strains containing a functional atoC gene. Although the exact mechanism of control was not evident from these studies, the data suggest that the atoC gene product is an activator which is required for the synthesis or activation of the atoDAB-encoded enzymes. PMID- 3025187 TI - Nitrogen fixation ability of exopolysaccharide synthesis mutants of Rhizobium sp. strain NGR234 and Rhizobium trifolii is restored by the addition of homologous exopolysaccharides. AB - Several transposon Tn5-induced mutants of the broad-host-range Rhizobium sp. strain NGR234 produce little or no detectable acidic exopolysaccharide (EPS) and are unable to induce nitrogen-fixing nodules on Leucaena leucocephala var. Peru or siratro plants. The ability of these Exo- mutants to induce functioning nodules on Leucaena plants was restored by coinoculation with a Sym plasmid-cured (Nod- Exo+) derivative of parent strain NGR234, purified EPS from the parent strain, or the oligosaccharide from the EPS. Coinoculation with EPS or related oligosaccharide also resulted in formation of nitrogen-fixing nodules on siratro plants. In addition, an Exo- mutant (ANU437) of Rhizobium trifolii ANU794 was able to form nitrogen-fixing nodules on white clover in the presence of added EPS or related oligosaccharide from R. trifolii ANU843. These results demonstrate that the absence of Rhizobium EPSs can result in failure of effective symbiosis with both temperate and subtropical legumes. PMID- 3025188 TI - The signal sequence suffices to direct export of outer membrane protein OmpA of Escherichia coli K-12. AB - We studied whether information required for export is present within the mature form of the Escherichia coli 325-residue outer membrane protein OmpA. We had previously analyzed overlapping internal deletions in the ompA gene, and the results allowed us to conclude that if such information exists it must be present repeatedly within the membrane part of the protein encompassing amino acid residues 1 to 177 (R. Freudl, H. Schwarz, M. Klose, N. R. Movva, and U. Henning, EMBO J. 4:3593-3598, 1985). A deletion which removed the codons for amino acid residues 1 to 229 of the OmpA protein was constructed. In this construct the signal sequence was fused to the periplasmic part of the protein. The resulting protein, designated Pro-OmpA delta 1-229, was processed, and the mature 95 residue protein accumulated in the periplasm. Hence, information required for export does not exist within the OmpA protein. PMID- 3025189 TI - Identification of transposable elements which activate gene expression in Pseudomonas cepacia. AB - This study demonstrated that transposable elements in Pseudomonas cepacia could be inserted upstream of a poorly expressed gene and increase its expression more than 30-fold. Five elements, TnPc1, IS402, IS403, IS404, and IS405, were isolated by their ability to increase expression of the beta-lactamase gene of the broad host-range plasmid pRP1. Increased expression resulted only from insertion of these elements, suggesting that insertional activation is an important means of elevating gene expression in this organism. Four of the elements inserted between a PstI site within the beta-lactamase gene and a BamHI site located 375 base pairs upstream of its promoter. The element IS403 inserted distal to the BamHI site within the coding region for the gene tnpR, suggesting that insertional activation can act over greater than expected distances. In addition, the element IS402 activated the beta-lactamase genes carried on plasmids pRP1 and pMR5 (temperature-sensitive pRP1) equally well in opposite orientations, demonstrating that insertional activation by this element occurs independent of its orientation. PMID- 3025191 TI - Effect of oxygen limitation on the formation of the electron transport system of the phytopathogenic fluorescent bacterium Pseudomonas cichorii. AB - The composition of the membrane-bound electron transport system of the phytopathogenic bacterium Pseudomonas cichorii underwent modification in response to oxygen supply. Growth adaptation to low oxygen concentrations was characterized by repression of cytochromes involved in ubiquinol-cyt. c oxidoreductase and cyt. c oxidase activities. By contrast, cyto. o, i.e., the alternative cyanide-insensitive oxidase of P. cichorii, was unaffected by low oxygen tension. No a-type cytochromes could be detected at any stage of growth. PMID- 3025190 TI - Cloning and analysis of transcription of the mei2 gene responsible for initiation of meiosis in the fission yeast Schizosaccharomyces pombe. AB - We have isolated a hybrid plasmid, pDB(mei2)2, containing a 7.4-kilobases (kb) DNA fragment from a Schizosaccharomyces pombe genomic library which is able to complement the mei2 mutation of S. pombe. Integration of the cloned DNA sequence at the mei2 site on chromosome I demonstrated that it contained the mei2 gene. This gene was localized on a 4.7-kb HindIII-PvuII fragment in the subclone pFMV402. Transcriptional regulation was studied by Northern blot analysis in which polyadenylated RNA was prepared from a heterozygous (h+N/h-S) diploid strain cultured either in nitrogen-rich growth medium or in nitrogen-free sporulation medium. The size of the major mei2 mRNA, which always gave a broad band, was estimated to be 4.2 +/- 0.2 kb, and a few minor bands (e.g., 3.2 and 1.8 kb) appeared as well. These transcripts appeared more abundantly in sporulating cells than in growing cells. Neither the mating type genes (mat) nor the mei3 gene was essential for transcription of the mei2 gene, since ample mei2 mRNA was detected in sporulation-deficient cells transferred to sporulation medium, such as h+N/h+N and h-S/h-S homozygotes, as well as mei1 and mei3 mutants. PMID- 3025193 TI - Cation-induced aggregation of membrane vesicles isolated from vascular smooth muscle. AB - Cations stimulated aortic muscle membrane aggregation with increasing potency according to their effective charge, e.g., K+ less than Mg2+ less than La3+, and the stimulation is reciprocally related to the apparent affinity for these cations. Divalent metal ion-induced membrane aggregation showed a dependence on the ionic radius, being optimal for Cd2+. Polyvalent cation-induced membrane aggregation was reversibly suppressed by high ionic strength as well as by metal ion chelators, irreversibly inhibited by the cross-linking agent glutaraldehyde, and enhanced by increasing concentrations of ethanol and increased temperature of the medium. When the pH is lowered below 6.0, membrane aggregation progressively increased with a concomitant decrease in cation-induced aggregation. The patterns of aggregation of microsomal membranes and further purified plasma membranes were almost identical whereas the aggregation of the heterogeneous mitochondrial membrane-enriched fraction was distinctly different in the initial rate of aggregation, its pH dependence, and metal ion concentration dependence. Our results indicate that cation-induced membrane aggregation can also be used to isolate a plasma membrane-enriched fraction from vascular smooth muscle. PMID- 3025192 TI - Transplasmalemma electron transport is changed in simian virus 40 transformed liver cells. AB - Transplasma membrane electron transport activity by fetal rat liver cells (RLA209 15) infected with a temperature-sensitive strain of SV40 has been measured with cells grown at the restrictive temperature (40 degrees C) and permissive temperature (33 degrees C). The transformed cells grown at 33 degrees C had only one-half the rate of external ferricyanide reduction as the nontransformed cells held at 40 degrees C. Both the Km and Vmax for ferricyanide reduction were changed in the transformed state. The change in Vmax can be based on a decrease of NADH in the transformed cells. The change in rate with ferricyanide does not depend on change in surface charge. Reduction of external ferricyanide was accompanied by release of protons from the cells. The ratio of protons released to ferricyanide reduced was higher in the transformed cells than in the non transformed cells. Since the transplasma membrane electron transport has been shown to stimulate cell growth under limiting serum, the changes in the plasma membrane electron transport and proton release in transformed cells may relate to modification of growth control. PMID- 3025194 TI - Fluoride and beryllium interact with the (Na + K)-dependent ATPase as analogs of phosphate. AB - Fluoride irreversibly inhibits the (Na + K)-ATPase, and this inactivation requires divalent cations (Mg2+, Mn2+, or Ca2+), is augmented by K+, but is diminished by Na+ and by ATP. Prior incubation with the aluminum chelator deferoxamine markedly slows inactivation, whereas adding 1 microM AlCl3 speeds it, consistent with AlF-4 being the active species. Prior incubation of the enzyme with vanadate also blocks inactivation by fluoride added subsequently. Fluoride stimulates ouabain binding to the enzyme, and thus the analogy between AlF-4 and both orthophosphate and orthovanadate is reflected not only in the similar dependence on specific ligands for their enzyme interactions and their apparent competition for the same sites, but also in their common ability to promote ouabain binding. Beryllium also irreversibly inhibits the enzyme, and this inactivation again requires divalent cations, is augmented by K+, but is diminished by Na+ and by ATP. Similarly, prior incubation of the enzyme with vanadate blocks inactivation by beryllium added subsequently. Inactivation by beryllium, however, does not require a halide, and, unlike inactivation by fluoride, increases at basic pHs. These observations suggest that beryllium, as beryllium hydroxide complexes, acts as a phosphate analog, similar to AlF-4 and vanadate. PMID- 3025196 TI - Two different phosphorylation-dephosphorylation cycles of Na,K-ATPase proteoliposomes accompanying Na+ transport in the absence of K+. AB - The phosphorylated intermediate (EP) of the Na,K-ATPase proteoliposomes (PL) prepared from the electric eel enzyme is composed of an ADP-sensitive K+ insensitive form (E1P), an ADP- and K+-sensitive form (E*P), and a K+-sensitive ADP-insensitive form (E2P). The composition of the intermediate varied with the cholesterol content of the lipid bilayer. The PL containing less than 30 mol % cholesterol (LCPL) formed E2P-rich EP in the presence of 10 mM Na+ on both sides at 15 degrees C, while the PL containing more than 35 mol % cholesterol (HCPL) formed E*P-rich EP under the same condition. In the presence of ionophore (monensin, nigericin, A23187), the HCPL formed E2P-rich EP as reported in the preceding paper. The turnover rate of Na-ATPase activity (the ratio of Na-ATPase to the EP level) in the LCPL was lower than that in the HCPL, and the addition of 20 microM monensin or A23187 to the HCPL reduced the Na-ATPase activity. The coupling ratio of Na+ influx (cellular efflux):Na+ efflux (cellular influx):ATP hydrolysis was 2.8:1.8:1 in the LCPL, although 1.6:0.6:1 in the HCPL. The coupling ratio of Na+ influx:ATP hydrolysis in the HCPL increased to 2.8:1 in the presence of A23187. Moreover, the increase of ATP concentration enhanced not only the Na-ATPase activity in the LCPL and HCPL with monensin but also the Na+ influx in the LCPL. This ATP enhancement was not found, however, in the HCPL without ionophores. The ADP enhancement of the Na+ influx was not observed in either the HCPL or the LCPL. We conclude from these observations that there are at least two different phosphorylation-dephosphorylation cycles (an E2P cycle and an E*P cycle) in the PL in the absence of K+. The E2P cycle transports three Na+ from the extravesicular (cytoplasmic) to the intravesicular (extracellular) side and two Na+ in the opposite direction per cycle and is similar to the ATP-dependent Na+-Na+ exchange system already reported (Blostein, R. (1983) J. Biol. Chem. 258, 7948-7953; Cornelius, F., and Skou, J. C. (1985) Biochim. Biophys. Acta 818, 211 221). However, the E*P cycle transports one Na+ from the extravesicular to the intravesicular side/cycle and has not yet been previously reported. PMID- 3025195 TI - Phosphorylated intermediates of Na,K-ATPase proteoliposomes controlled by bilayer cholesterol. Interaction with cardiac steroid. AB - Fragmental Na,K-ATPase from the electric eel forms three phosphorylated intermediates (EP) with MgATP and Na+: ADP-sensitive K+-insensitive EP (E1P), ADP and K+-sensitive EP (E*P), and K+-sensitive ADP-insensitive EP (E2P). The EP composition varied with the Na+ concentration. In the reconstituted Na,K-ATPase proteoliposomes (PL), the EP composition of the inside-out form was controlled not only by the intravesicular (extracellular) Na+ concentration, but also by the temperature and the cholesterol content of the lipid bilayer. When the lipid bilayer of PL contained less than 30 mol % cholesterol, the E*P content did not change significantly while the E2P content increased with an elevation in temperature (3-20 degrees C). In contrast, when the lipid bilayer contains more than 35 mol % cholesterol, the E*P content increased while the E2P content stayed less than 10% under the same temperature change. These observations suggest that a high cholesterol content in the lipid bilayer interferes with the E*P to E2P conversion. This cholesterol effect was reversed by ionophores (monensin, nigericin, and A23187). Therefore, E1P-rich EP, E*P-rich EP, or E2P-rich EP could be obtained in the PL under a constant Na+ concentration by using various concentrations of cholesterol and ionophores. The reaction between the proteoliposomal EPs and digitoxigenin (lipid-soluble cardiac steroid) occurred in a single turnover, thereby avoiding unphysiologically high Na+ concentrations. The increase in the ADP- and K+-insensitive EP, which indicated formation of the digitoxigenin-Na,K-ATPase complex, was equivalent to the decrease in the E*P under six different sets of conditions, without any significant change in the E1P and E2P contents. This result indicated that E*P is the active intermediate of the Na,K-ATPase for cardiac steroid binding. Although the E2P has been thought to be the active form for binding, it cannot bind with the cardiac steroid in the presence of Na+ and in the absence of free Mg2+. PMID- 3025197 TI - Drosophila deoxyuridine triphosphatase. Purification and characterization. AB - Deoxyuridine triphosphatase (dUTPase), an enzyme that catalyzes hydrolysis of dUTP to deoxyuridylate and inorganic pyrophosphate, has been purified approximately 6,000-fold from Drosophila embryos. The enzyme has a native molecular weight of 46,000 and a sedimentation coefficient of 3.5 S. The enzyme is most likely a metalloenzyme. It is specific for dUTP among the DNA nucleotides tested, with an apparent Km of 1 microM. The expression of dUTPase appears stage specific, with embryos representing the only step in the life cycle of Drosophila with clearly detectable levels of the enzyme. While other possibilities exist, these results suggest an enhanced opportunity for the inclusion of uracil into Drosophila DNA subsequent to embryonic development. PMID- 3025198 TI - Calcium and the phosphoinositide cycle in WRK-1 cells. Effects of A23187 on metabolism of specific phosphatidylinositol pools. AB - WRK-1 cells possess a labile, hormone-sensitive pool of phosphatidylinositol which appears to be separate from the stable, hormone-insensitive phosphatidylinositol. It is the sensitive pool which turns over in response to treatment with vasopressin. Addition of the calcium ionophore A23187, on the other hand, selectively stimulates precursor incorporation into the hormone insensitive pool of phosphatidylinositol, while causing nonspecific breakdown of both pools. The polyphosphoinositides are similarly affected. Ionophore stimulated breakdown appears to be predominantly phospholipase C-mediated, since there is a concomitant increase in inositol phosphates. These inositol phosphates are localized predominantly in the extracellular medium. Permeabilization of the cells may explain the extracellular location of the breakdown products. When added together with the hormone, A23187, at concentrations greater than 5 X 10( 6) M, inhibits both hormone-induced synthesis and breakdown of phosphatidylinositol. Omission of calcium from the medium abolishes the effects of the ionophore. PMID- 3025199 TI - In vitro expression of the Escherichia coli nusA-infB operon. AB - The expression of the nusA-infB operon has been investigated using an in vitro system based on the formation of the first dipeptide of the gene product. A series of plasmids containing various deletions of the operon were used as templates in this study. Of the four genes coding for protein products, 15Ka, nusA, infB, and 15Kb, only 15Ka was not expressed in this dipeptide system. The initial dipeptides for the other gene products, fMet-Asn (pnusA), fMet-Thr (IF-2 alpha), and fMet-Ala (p15Kb), were synthesized even from plasmids lacking the primary promoters. It appears that secondary (internal) promoters in the operon can efficiently direct the expression of these genes. No regulation of the expression was observed with IF-2 alpha, but pnusA inhibited the expression of the nusA gene (autoregulation) as well as the p15b gene. Experiments using an uncoupled system indicated that the effect of pnusA on nusA expression was at the level of transcription, but that both a transcriptional and a post transcriptional effect of pnusA was seen on 15Kb expression. PMID- 3025200 TI - Characterization of receptors for vasoactive intestinal peptide solubilized from the lung. AB - The zwitterionic detergent CHAPS was used to solubilize functional receptors for vasoactive intestinal peptide (VIP) from guinea pig lung. The solubilized receptors were resolved by high performance gel filtration in 3 mM CHAPS into two active fractions with apparent Stokes radii of 5.9 +/- 0.1 and 2.3 +/- 0.1 nm. The binding of 125I-VIP to the two receptor fractions was time-dependent, reversible, and saturable. Trypsin destroyed the binding activity of the receptor fractions, indicating their proteinic nature. Unlabeled VIP competitively displaced the binding of 125I-VIP to the 5.9-nm fraction (IC50 = 240 pM) and the 2.3-nm fraction (IC50 = 1.2 microM). Scatchard analysis indicated a single class of binding sites in each receptor fraction, with Kd values 300 pM and 0.97 microM for the 5.9- and 2.3-nm Stokes radii fractions, respectively. When the high affinity, 5.9-nm Stokes radius fraction was rechromatographed in 9 nM CHAPS, 46% of the binding activity eluted in the low affinity, 2.3-nm Stokes radius fraction, indicating that the latter is a product of dissociation of the high affinity receptor complex. GTP inhibited the binding of 125I-VIP to the high affinity complex but not the low affinity species. Scatchard plots of VIP binding by the high affinity receptors treated with GTP suggested the presence of two distinct binding sites (Kd 4.4 and 153 nM), compared to a single binding site (Kd = 0.3 nM) obtained in untreated receptors. The nonhydrolyzable GTP analog, guanyl 5'-yl-imidodiphosphate, inhibited VIP binding by the high affinity receptor fraction with potency nearly equivalent to that of GTP. These observations suggest that GTP-binding regulatory proteins are functionally coupled to the VIP binding subunit in the high affinity receptor complex. The peptide specificity characteristics of the two receptor fractions were different. Peptide histidine isoleucine and growth hormone releasing factor, peptides homologous to VIP, were 87.5- and 22.9-fold less potent than VIP in displacing 125I-VIP binding by the high affinity receptor complex, respectively. On the other hand, growth hormone releasing factor was more potent (22.7-fold) and peptide histidine isoleucine was less potent (31.3-fold) than VIP in displacing the binding by the low affinity species. PMID- 3025201 TI - Light activation of phospholipase A2 in rod outer segments of bovine retina and its modulation by GTP-binding proteins. AB - Light stimulates phospholipase A2 activity in rod outer segments (ROS) of bovine retina as measured by the liberation of arachidonate from phosphatidylcholine, in in vitro assays of dark-adapted ROS. A role for GTP-binding proteins (G or N proteins) in the light activation of phospholipase A2 is suggested by the capacity for guanosine 5'-O-(thiotriphosphate) (GTP gamma S) to activate phospholipase A2 in dark-adapted ROS. In contrast, addition of GTP gamma S coincident with light exposure inhibited the light activation of phospholipase A2, suggesting that phospholipase A2 activity in the ROS is under dual regulation by G proteins. Transducin, the major G protein of the ROS, mediates the activation of cGMP phosphodiesterase by light and is a substrate for both cholera and pertussis toxin. Treatment of dark-adapted ROS with either toxin inhibits both basal and light-activated phospholipase A2, mimicking the action of the toxins on the light-induced cGMP phosphodiesterase activity of ROS. There is a loss of light-sensitive phospholipase A2 activity coincident with extraction of transducin from ROS membranes. In addition, the light-sensitive phospholipase A2 activity can be partially restored by the addition of purified transducin to the extracted ROS membranes. Light activation of phospholipase A2 in ROS membranes thus occurs by a transducin-dependent mechanism. PMID- 3025202 TI - The effects of bombesin on polyphosphoinositide and calcium metabolism in Swiss 3T3 cells. AB - Bombesin, a peptide mitogen for a variety of cell types, acts as a typical Ca2+ mobilizing hormone in Swiss 3T3 fibroblasts. At its mitogenic concentrations (1 25 nM), bombesin stimulates polyphosphoinositide turnover, i.e. breakdown of phosphatidylinositol 4,5-bisphosphate and a concomitant increase in inositol phosphates in a time- and dose-dependent manner. In particular, bombesin induces an initial transient increase in inositol 1,4,5-trisphosphate concentration, followed by an increase in the concentration of inositol 1,3,4-trisphosphate. Also, within 30 s of bombesin addition, the mass of 1,2-diacylglycerol nearly doubles and remains at this level for up to 60 min. Intracellular [Ca2+] measurements with a photoprotein, aequorin, demonstrate that bombesin stimulates a transient rise in cytosolic free Ca2+ concentration. A mobilization of Ca2+ from an intracellular pool is observed as a dose-dependent, transient increase in 45Ca2+ efflux from prelabeled cells, both in the presence and absence of extracellular Ca2+. Bombesin also induces a sustained increase in Ca2+ influx rate and stimulates 3-O-methyl-D-glucose transport across the plasma membrane. These composite results indicate that the mitogenic effect of bombesin is mediated through an activation of the Ca2+ messenger system. PMID- 3025203 TI - Crucial role of the connecting region joining the two functional domains of yeast tryptophan synthetase. AB - We constructed a hybrid plasmid expressing yeast tryptophan synthetase in Escherichia coli. Several deletion variants lacking the A or B domains of this polypeptide (recognized by their homology to the alpha and beta subunits of prokaryotic tryptophan synthetase) showed no enzymatic activity and failed to substitute for the corresponding E. coli subunits. To examine the role of a presumed interdomain connecting region in the yeast enzyme, we constructed a variant lacking 18 amino acids in that region. The variant polypeptide was completely inactive. Replacing 14 of the 18 missing amino acids with a segment having a different sequence partially restored activity. A spontaneous revertant was characterized and shown to have a duplication of 16 amino acid residues in this region; the activity of the duplication polypeptide was better than that of the 14-residue replacement. If confirmed by additional studies, our finding that the length of the connecting region is more critical than its sequence has implications for understanding the origin of gene fusions during evolution as well as for designing artificial fusions. PMID- 3025204 TI - Regulation of insulin-stimulated glucose transport in the isolated rat adipocyte. cAMP-independent effects of lipolytic and antilipolytic agents. AB - This paper examines the modulation of insulin-stimulated glucose transport activity in rat adipose cells by ligands for receptors (R) that mediate stimulation (Rs; lipolytic) or inhibition (Ri; antilipolytic) of adenylate cyclase. The changes in glucose transport activity and cAMP, as assessed by 3-O methylglucose uptake and (-/+) cAMP-dependent protein kinase (A-kinase) activity ratios, respectively, were monitored under conditions that maintain steady-state A-kinase activity ratios (Honnor, R. C., Dhillon, G. S., and Londos, C. (1985) J. Biol. Chem. 260, 15122-15129). Removal of endogenous adenosine with adenosine deaminase decreased insulin-stimulated glucose transport activity by approximately 30%, which was prevented or restored with Ri agonists such as phenylisopropyladenosine, nicotinic acid, and prostaglandin E1. These changes in transport activity were not accompanied by changes in A-kinase activity ratios, indicating that Ri-mediated effects on transport are independent of cAMP changes. Addition of an Rs ligand, isoproterenol, in the presence of adenosine increased kinase activity but did not change glucose transport activity. Conversely, upon removal of adenosine, addition of Rs ligands such as isoproterenol, adrenocorticotropic hormone, or glucagon strongly inhibited transport (approximately 50%) and stimulated kinase activity. However, subsequent addition of phenylisopropyladenosine nearly restored transport activity without alteration of A-kinase activity. These data and additional kinetic experiments suggest that Rs-mediated glucose transport modulations are also independent of cAMP. The interchangeability of ligands for both Rs and Ri receptors in modulating transport activity suggests that these cAMP-independent effects are mediated by the stimulatory (Ns) and inhibitory (Ni) guanyl nucleotide-binding regulatory proteins of adenylate cyclase. All Rs-and Ri-induced changes in transport activity occurred without a change in glucose transporter distribution, as assessed by D-glucose-inhibitable cytochalasin B binding, suggesting that Rs and Ri ligands modulate the intrinsic activity of the glucose transporter present in the plasma membrane. PMID- 3025205 TI - The role of nicotinamide adenine dinucleotide in the inhibition of bovine liver S adenosylhomocysteine hydrolase by neplanocin A. AB - Neoplanocin A, a cyclopentenyl analog of adenosine, has been shown recently to be a tight binding inhibitor of S-adenosylhomocysteine (AdoHcy) hydrolase (EC 3.3.1.1), exhibiting a stoichiometry of one molecule of inhibitor per molecule of the enzyme tetramer (Borchardt, R. T., Keller, B. T., and Patel-Thombre, U. (1984) J. Biol. Chem. 259, 4353-4358). In the present study a detailed analysis was performed of the possible role of the enzyme-bound NAD+ in the inactivation of AdoHcy hydrolase by neplanocin A. The NAD+/NADH content was quantitated using a fluorescence technique. The native enzyme showed intrinsic fluorescence with an emission maximum at 460 nm when excited at 340 nm, partially due to NADH bound to the enzyme. It was found that the content of NAD+ and NADH in freshly prepared, native enzyme is equal, having a stoichiometry of two nucleotides per enzyme molecule (tetramer). In addition, it was observed that the enzymatic activity of the native enzyme can be increased by about 30% following preincubation with NAD+. Furthermore, it was demonstrated that the mechanism of inhibition of AdoHcy hydrolase by neplanocin A involves the reduction of enzymatically bound NAD+ to NADH. Catalytic activity of the inactivated enzyme could be fully recovered in a time-dependent manner by further incubation with NAD+ (but not NADH). It was also found that inhibition by neplanocin A does not involve dissociation of the bound NAD+ or NADH from the enzyme, but simply reduction of the NAD+ to NADH. PMID- 3025206 TI - Regulation of glycogen synthase activity by growth factors. Relationship between synthase activation and receptor occupancy. AB - Platelet-derived growth factor (PDGF) was found to stimulate the activity of glycogen synthase, an enzyme subjected to regulation by phosphorylation dephosphorylation reactions. In Swiss mouse 3T3 cells, the time course of enzyme activation by PDGF is very similar to that of epidermal growth factor (EGF) and insulin. A 3-fold maximal stimulation was observed by 30 min, and the enzyme activity ratio returned to basal levels by 100 min. The PDGF effect was maximal at 1 ng/ml (30 pM) and half-maximal stimulation was observed at 0.2 ng/ml (6 pM). Parallel measurements of 125I-PDGF binding indicate that binding was maximal by 10 min and thus should not be rate-limiting for enzyme activation. In addition, presaturation of the receptors with PDGF at 4 degrees C did not expedite subsequent enzyme activation at 37 degrees C. Removal of PDGF in the media after the 4 degrees C pretreatment did not affect the enzyme activation response, indicating that the continued presence of PDGF in the medium is not necessary after receptor binding is saturated. Results of sequential addition of PDGF, EGF, and insulin indicated a refractory period in the response system. This property is evident even when the second addition involved a different growth factor and is independent of the sequence of addition of the factors. There was little additivity in the actions of the three growth factors in effecting enzyme activation and suggests a common intermediate element in the three signalling pathways. PMID- 3025207 TI - Agents that elevate cAMP levels in platelets decrease thrombin binding. AB - The effect of high intracellular levels of cAMP on the ability of rabbit and human platelets to bind and respond to thrombin was examined. Control rabbit platelets differed from human platelets in two interesting respects: they showed thrombin-dependent up-regulation of thrombin binding, but also a 3- to 5-fold lower thrombin-binding capacity. Nevertheless, treatment with prostaglandin E1 + theophylline or with forskolin decreased thrombin binding to both rabbit and human platelets by 60 to 70%. This effect was associated with a marked increase in the level of cAMP and seemed to depend on a decrease in number rather than affinity of thrombin-binding sites. Changes in thrombin binding correlated closely with changes in thrombin-stimulated incorporation of 32Pi into phosphatidic acid and a 40-kDa protein. However, regardless of the amount of thrombin that bound to treated platelets, thrombin-stimulated phosphorylation of a 20-kDa protein and serotonin secretion were severely inhibited. Thus, increased levels of platelet cAMP are associated with a reduced ability to bind and respond to thrombin. However, thrombin binding to platelets correlates more closely with some responses than others, presumably because cAMP inhibits additional platelet reactions. PMID- 3025208 TI - Ascaris suum phosphofructokinase. Phosphorylation by protein kinase and sequence of the phosphopeptide. AB - Phosphorylation of the ascarid phosphofructokinase with the catalytic subunit of beef heart cyclic AMP-dependent protein kinase results in the incorporation of 1 mol of P/mol of subunit. Accompanying the phosphorylation there is a 3-4-fold increase in catalytic activity when measured at pH 6.8 with inhibitory levels of ATP. Studies on the effect of phosphorylation on the ATP saturation curve demonstrated that phosphorylation decreased the inhibitory action of ATP. The apparent Km of the catalytic subunit for the phosphofructokinase was 11.2 microM. Chymotryptic or subtilisin digestion of the labeled enzyme released distinct but overlapping phosphopeptides that were purified by high pressure liquid chromatography and sequenced by gas phase peptide sequencing. The sequence of the chymotryptic peptide was Ala-Lys-Gly-Arg-Ser-Asp-Ser(P)-Ile-Val-Pro-Thr. Based on these results and earlier observations, it is proposed that phosphorylation of phosphofructokinase plays an important role in the regulation of energy metabolism in the parasitic helminth. PMID- 3025209 TI - Short-term feedback regulation of cAMP by accelerated degradation in rat tissues. AB - A recent study showed that cAMP analogs lowered cAMP levels in rat hepatocytes (Corbin, J.D., Beebe, S.J., and Blackmore, P.F. (1985) J. Biol. Chem. 260, 8731 8735). The present work demonstrates that cAMP analogs also lowered cAMP in a rapid, concentration-dependent manner in heart and fat cells. In order to determine if the cAMP-dependent protein kinase mediated this effect, techniques were developed to assay the protein kinase activity ratio in hepatocytes treated with cAMP analogs. The activation of protein kinase and phosphorylase in hepatocytes by 8-pCl phi S-cAMP (where 8-pCl phi S- indicates 8 parachlorothiophenyl-) was concentration-dependent and occurred in parallel to proportionate decreases in cAMP. More than 20% of the cAMP binding sites on the protein kinase were unoccupied at concentrations of 8-pCl phi S-cAMP that produced maximal cAMP lowering. Thus, the possibility that 8-pCl phi S-cAMP lowered cAMP by displacing it from protein kinase binding sites, making it available for hydrolysis, seemed unlikely. In adipocytes, the lowering of cAMP by 8-pCl phi S-cAMP occurred in parallel with increases in lipolysis and activation of low Km phosphodiesterase, suggesting that the phosphodiesterase was responsible for the cAMP lowering. Further evidence for this assertion was the finding that in hepatocytes preloaded with low concentrations of 8-pCl phi S cAMP, glucagon lowered 8-pCl phi S-cAMP by about 50%, an amount similar to the cAMP lowering observed with 8-pCl phi S-cAMP treatment. The results were consistent with a cAMP-dependent protein kinase-catalyzed activation of a phosphodiesterase and suggested that 8-pCl phi S-cAMP-mediated hydrolysis of cAMP mimicked a physiologically significant response. The observation of this phenomenon in several tissues further suggested that it may be a general mechanism for dampening and terminating the hormonal signal through accelerated degradation of cAMP. PMID- 3025210 TI - The dynamics of ubiquitin pools within cultured human lung fibroblasts. AB - The dynamics of ubiquitin pools within the cultured human lung fibroblast line IMR-90 were examined using solid phase immunochemical methods to quantitate free and conjugated polypeptide. Fetal calf serum was found to contain a nondialyzable factor that induced a transient accumulation of ubiquitin. During the induction, free and conjugated ubiquitin pools changed in concert so that the fraction conjugated remained constant. The induction of ubiquitin by the serum factor resulted from an enhanced rate of protein synthesis. Within experimental error no change in the first order rate constant for intracellular ubiquitin degradation was observed. Pulse-chase studies revealed ubiquitin to turn over with a half life of 28-31 h in conditioned and freshly fed cultures. Withdrawal of serum from cultures led to a rapid decline in total ubiquitin during which the fractional level of conjugation remained constant. The accelerated ubiquitin turnover following removal of serum likely involves lysosomal autophagy since 10 mM NH4Cl led to an accumulation of the polypeptide. Since no similar effect of the lysosomotropic compound was observed in conditioned or freshly fed cultures, nonlysosomal processes are probably responsible for ubiquitin turnover under nutritional balance. The dynamics of these intracellular pools suggests that the ubiquitin ligation system is subject to regulatory constraints not previously suspected. The short half-life for ubiquitin is consistent with the apparent ability of cells to alter ubiquitin levels in response to external stimuli and stress. PMID- 3025211 TI - Purification of the surface cAMP receptor in Dictyostelium. AB - We have previously identified and demonstrated reversible ligand-induced modification of the major cell surface cAMP receptor in Dictyostelium discoideum. The receptor, or a subunit of it, has been purified to homogeneity by hydroxylapatite chromatography followed by two-dimensional preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The purification was monitored by following 32Pi incorporated by photoaffinity labeling with 8-azido [32P]cAMP or by in vivo labeling with 32Pi. Two interconvertible forms of the receptor, designated R (Mr 40,000) and D (Mr 43,000), co-purified. Two dimensional peptide maps of independently purified and 125I-iodinated R and D forms of the receptor were nearly identical but did have several distinct peptides. The estimated 6000-fold purification required is consistent with the number of cell surface binding sites assuming there are not multiple binding sites/polypeptide. In the accompanying article we report the generation of a monospecific polyclonal antiserum which has helped to further elucidate the physical properties and developmental regulation of the cAMP receptor. PMID- 3025212 TI - The surface cyclic AMP receptor in Dictyostelium. Levels of ligand-induced phosphorylation, solubilization, identification of primary transcript, and developmental regulation of expression. AB - A monospecific polyclonal antiserum to the surface cAMP receptor of Dictyostelium has been developed by immunization with purified receptor immobilized on particles of polyacrylamide and on nitrocellulose paper. In Western blots, the antiserum displays high affinity and specificity for both the R (Mr 40,000) and D (Mr 43,000) forms of the receptor previously identified by photoaffinity labeling with 8-azido-[32P] cAMP. These bands, labeled with the photoaffinity label or with 32 Pi, were quantitatively and specifically immunoprecipitated, supporting co-purification data that all represent the same polypeptide. The R form, found in unstimulated cells, contained at least 0.2 mol of phosphate/mol of receptor. The D form, generated by cAMP stimulation of intact cells, contained at least 4 mol of phosphate/mol of receptor. In the absence of detergents, the receptor was exclusively located on membranes. The receptor was solubilized effectively in Triton X-100 and sedimented as a broad peak of 5-7 S on sucrose velocity gradients. Western blots of membranes isolated at different times after starvation indicate that the appearance of cell surface cAMP binding sites during the aggregation stage of development (5-6 h) is due to de novo synthesis of receptor protein. Pulse labeling with [35S]methionine indicated that the receptor is most rapidly synthesized during the preaggregation stage of development (1-3 h), prior to its maximal accumulation in membranes. The serum specifically immunoprecipitates a polypeptide of Mr 37,000 from an in vitro translation reaction using RNA isolated from preaggregation stage cells. The time course of expression of the mRNA coding for the Mr 37,000 polypeptide parallels the rate of receptor synthesis in vivo. PMID- 3025213 TI - Mossbauer study of Clostridium pasteurianum hydrogenase II. Evidence for a novel three-iron cluster. AB - Hydrogenase II contains two iron-sulfur clusters, one of the [4Fe-4S] type and one of unknown structure with unusual spectral properties (H-cluster). Using Mossbauer spectroscopy we have studied the H-cluster under a variety of conditions. In the reduced state the cluster exhibits, in zero magnetic field, spectra with the typical 2:1 quadrupole pattern of reduced [3Fe-4S] clusters. However, whereas the latter are paramagnetic (S = 2) the H-cluster is diamagnetic (S = 0). Upon oxidation and exposure to CO the H-cluster exhibits an S = 1/2 EPR spectrum with g values at 2.03, 2.02, and 2.00. In this state, the Mossbauer spectra reveal two cluster subsites with magnetic hyperfine coupling constants AI = +26.5 MHz and AII = -30 MHz. ENDOR data obtained by Hoffman and co-workers (Telser, J., Benecky, M. J., Adams, M. W. W., Mortenson, L. E., and Hoffman, B. M. (1986) J. Biol. Chem. 261, 13536-13541) show a 57Fe resonance at AIII approximately equal to 9.5 MHz. Analysis of the Mossbauer spectra shows that this resonance represents one iron site. Our studies of the reduced and CO-bound oxidized states of hydrogenase II suggest that the H-cluster contains three iron atoms. The data obtained for the oxidized H-cluster suggest a novel type of 3-Fe cluster and bear little resemblance to those reported for oxidized [3Fe-4S] clusters with g = 2.01 EPR signals. In the reduced sample the [4Fe-4S]1+ cluster appears to occur in a mixture of two distinct electronic states. PMID- 3025214 TI - The Lebanese allele at the low density lipoprotein receptor locus. Nonsense mutation produces truncated receptor that is retained in endoplasmic reticulum. AB - We here describe a mutant low density lipoprotein receptor gene that produces a shortened receptor protein lacking three domains: the region of clustered O linked carbohydrates, the membrane-spanning region, and the cytoplasmic tail. The defect is attributable to a single nucleotide substitution that creates a premature termination codon at amino acid 660, eliminating 180 residues from the mature protein. The truncated protein retains only two domains: a complete ligand binding region (residues 1-292) and a partial epidermal growth factor precursor homology region (residues 293-659). The termination codon occurs in the middle of a cysteine-rich sequence that is part of the epidermal growth factor precursor homology domain. The mutant protein is present in markedly reduced amounts and may be translated at a reduced rate. After synthesis, most of the receptor remains within the cell for several hours with its N-linked carbohydrate in an unprocessed endoglycosidase H-sensitive form. This finding suggests that the shortened receptor leaves the endoplasmic reticulum at an abnormally slow rate, which is likely attributable to abnormal folding of the truncated protein. The mutation creates a new restriction site for the enzyme HinfI, thus permitting diagnosis by Southern blotting of genomic DNA. Two copies of this mutant gene were present in each of four unrelated Arab patients with homozygous familial hypercholesterolemia (three from Lebanon and one from Syria). We believe that this mutation, hereafter referred to as the "Lebanese allele," is responsible for the extraordinarily high incidence of familial hypercholesterolemia in Lebanon. PMID- 3025215 TI - Nuclear endo-exonuclease of Neurospora crassa. Evidence for a role in DNA repair. AB - The major nuclease activity in nuclei of mycelia of Neurospora crassa has been identified as that of endoexonuclease, an enzyme purified and characterized previously from mitochondria and vacuoles which acts endonucleolytically on single-stranded DNA and RNA and possesses highly processive exonuclease activity with double-stranded DNA. Cross-contamination from the other organelles was eliminated as a source of the activity. Endo-exonuclease of nucleoplasm, chromatin, and nuclear matrix showed 80-100% cross-reaction with antisera raised to purified extranuclear endoexonuclease and was also strongly inhibited by 20 microM aurin tricarboxylic acid. In addition, it yielded some of the same-sized polypeptides on activity gel analysis. Nuclei also contained immunochemically cross-reactive trypsin-activable endo-exonuclease activity, a form of enzyme that was shown previously to occur in high amounts in the cytosol and in a tightly bound form associated with the mitochondrial inner membrane. Pretreatment of wild type mycelia for 1 h with 4-16 micrograms/ml the DNA-damaging agent, 4 nitroquinoline-1-oxide (4-NQO), which caused about 50-80% growth inhibition, resulted in a dose-dependent loss of up to 80% of inactive endo-exonuclease from nuclei. At low doses of 4-NQO, this was accompanied by increases in the level of active enzyme. Nuclei of the DNA repair-deficient uvs-3 mutant were found to contain only 12% of the active enzyme and about 32% of inactive enzyme as that in wild-type nuclei. Mycelial growth of this mutant was 10 times more sensitive to 4 NQO than the wild-type. At a dose which resulted in equivalent growth inhibition, 4-NQO had no effect on the level of active endo-exonuclease in uvs-3 nuclei and caused an increase (over 30%) in the level of inactive enzyme. These data are consistent with a role of endo-exonuclease in the repair of nuclear DNA. PMID- 3025216 TI - Immunoprecipitation of the lutropin/choriogonadotropin receptor from biosynthetically labeled Leydig tumor cells. A 92-kDa Glycoprotein. AB - The structure of the lutropin/choriogonadotropin (LH/CG) receptor has been studied by immunoprecipitating the receptor from biosynthetically labeled cultured Leydig tumor cells (designated MA-10). This was performed by binding human choriogonadotropin (hCG) to the labeled cells, solubilizing the hormone receptor complex, partially purifying the complex by lectin chromatography, and immunoprecipitating the complex with an antibody that recognizes receptor-bound hCG. The conditions used for the release of the radiolabeled receptor from the immunoprecipitate and the subsequent analysis of this material on sodium dodecyl sulfate gels allowed us to determine directly the structure of the free (not hormone-occupied) LH/CG receptor. From experiments using cells labeled with [35S]methionine and [35S]cysteine, we show that the LH/CG receptor is composed of a single polypeptide chain that migrates as a 92-kDa protein on sodium dodecyl sulfate gels whether analyzed in the absence or presence of reducing agents. Other studies presented demonstrate that the LH/CG receptor is a glycoprotein. PMID- 3025217 TI - Hydroxyl radical formation and iron-binding proteins. Stimulation by the purple acid phosphatases. AB - The effect of the purple acid phosphatases with binuclear iron centers (uteroferrin and bovine spleen phosphatase) on hydroxyl radical formation by iron catalyzed Haber-Weiss-Fenton chemistry has been compared to that of lactoferrin and transferrin. Using 5,5-dimethyl-1-pyrroline-1-oxide to detect superoxide and hydroxyl radicals and the xanthine-xanthine oxidase system to generate superoxide and hydrogen peroxide, we have observed by ESR spectroscopy that both phosphatases were able to promote hydroxyl radical formation. Lactoferrin and transferrin were found incapable of giving rise to these reactive species. This can be explained by the fact that lactoferrin and transferrin carry two Fe(III) atoms per molecule, neither of which are readily reduced by biological reductants. In contrast, the phosphatases possess a binuclear iron center in which one of the iron atoms is stabilized in the ferric state, but the other freely undergoes one-electron redox reactions. The redox-active iron may act as a catalyst of the Haber-Weiss-Fenton sequence, thus enabling the reactions generating hydroxyl radical to proceed. The iron complex of diethylenetriamine penta-acetic acid, also redox active, was investigated and found as well to promote Haber-Weiss-Fenton chemistry. PMID- 3025218 TI - The sorting of proteins to the plasma membrane in epithelial cells. PMID- 3025219 TI - Topoisomerase II: A specific marker for cell proliferation. AB - We have used an antibody probe to measure the levels of topoisomerase II in several transformed and developmentally regulated normal cell types. Transformed cells contain roughly 1 X 10(6) copies of the enzyme. During erythropoiesis in chicken embryos the enzyme level drops from 7.8 X 10(4) copies per erythroblast to less than 300 copies per erythrocyte concomitant with the cessation of mitosis in the blood. Cultured myoblasts also lose topoisomerase II upon fusion into nonproliferating myotubes. When peripheral blood lymphocytes (which lack detectable topoisomerase II) commence proliferation, they express topoisomerase II de novo. Appearance of the enzyme exactly parallels the onset of DNA replication. These results suggest that topoisomerase II is not required for transcription in higher eukaryotes, but that it may function during DNA replication. Furthermore, topoisomerase II is a sensitive and specific marker for proliferating cells. PMID- 3025220 TI - Protein translocation across the yeast microsomal membrane is stimulated by a soluble factor. AB - We have found that a soluble activity present in the postribosomal supernatant fraction of Saccharomyces cerevisiae stimulates posttranslational translocation of yeast prepro-alpha-factor across yeast microsomal membranes. Stimulation of translocation is not due to a nonspecific affect on ATP levels. The activity is likely to be due to protein(s) as it is destroyed by N-ethylmaleimide, protease, or heat treatment but not by incubation with RNase. Its apparent sedimentation coefficient is approximately 9.6 S. PMID- 3025221 TI - Identification of an alternatively spliced site in human plasma fibronectin that mediates cell type-specific adhesion. AB - We have compared the molecular specificities of the adhesive interactions of melanoma and fibroblastic cells with fibronectin. Several striking differences were found in the sensitivity of the two cell types to inhibition by a series of synthetic peptides modeled on the Arg-Gly-Asp-Ser (RGDS) tetrapeptide adhesion signal. Further evidence for differences between the melanoma and fibroblastic cell adhesion systems was obtained by examining adhesion to proteolytic fragments of fibronectin. Fibroblastic BHK cells spread readily on fl3, a 75-kD fragment representing the RGDS-containing, "cell-binding" domain of fibronectin, but B16 F10 melanoma cells could not. The melanoma cells were able to spread instead on f9, a 113-kD fragment derived from the large subunit of fibronectin that contains at least part of the type III connecting segment difference region (or "V" region); f7, a fragment from the small fibronectin subunit that lacks this alternatively spliced polypeptide was inactive. Monoclonal antibody and fl3 inhibition experiments confirmed the inability of the melanoma cells to use the RGDS sequence; neither molecule affected melanoma cell spreading, but both completely abrogated fibroblast adhesion. By systematic analysis of a series of six overlapping synthetic peptides spanning the entire type III connecting segment, a novel attachment site was identified in a peptide near the COOH terminus of this region. The tetrapeptide sequence Arg-Glu-Asp-Val (REDV), which is somewhat related to RGDS, was present in this peptide in a highly hydrophilic region of the type III connecting segment. REDV appeared to be functionally important, since this synthetic tetrapeptide was inhibitory for melanoma cell adhesion to fibronectin but was inactive for fibroblastic cell adhesion. REDV therefore represents a novel adhesive recognition signal in fibronectin that possesses cell type specificity. These results suggest that, for some cell types, regulation of the adhesion-promoting activity of fibronectin may occur by alternative mRNA splicing. PMID- 3025222 TI - Distinct molecular interactions mediate neuronal process outgrowth on non neuronal cell surfaces and extracellular matrices. AB - We have compared neurite outgrowth on extracellular matrix (ECM) constituents to outgrowth on glial and muscle cell surfaces. Embryonic chick ciliary ganglion (CG) neurons regenerate neurites rapidly on surfaces coated with laminin (LN), fibronectin (FN), conditioned media (CM) from several non-neuronal cell types that secrete LN, and on intact extracellular matrices. Neurite outgrowth on all of these substrates is blocked by two monoclonal antibodies, CSAT and JG22, that prevent the adhesion of many cells, including neurons, to the ECM constituents LN, FN, and collagen. Neurite outgrowth is inhibited even on mixed LN/poly-D lysine substrates where neuronal attachment is independent of LN. Therefore, neuronal process outgrowth on extracellular matrices requires the function of neuronal cell surface molecules recognized by these antibodies. The surfaces of cultured astrocytes, Schwann cells, and skeletal myotubes also promote rapid process outgrowth from CG neurons. Neurite outgrowth on these surfaces, though, is not prevented by CSAT or JG22 antibodies. In addition, antibodies to a LN/proteoglycan complex that block neurite outgrowth on several LN-containing CM factors and on an ECM extract failed to inhibit cell surface-stimulated neurite outgrowth. After extraction with a nonionic detergent, Schwann cells and myotubes continue to support rapid neurite outgrowth. However, the activity associated with the detergent insoluble residue is blocked by CSAT and JG22 antibodies. Detergent extraction of astrocytes, in contrast, removes all neurite-promoting activity. These results provide evidence for at least two types of neuronal interactions with cells that promote neurite outgrowth. One involves adhesive proteins present in the ECM and ECM receptors on neurons. The second is mediated through detergent-extractable macromolecules present on non-neuronal cell surfaces and different, uncharacterized receptor(s) on neurons. Schwann cells and skeletal myotubes appear to promote neurite outgrowth by both mechanisms. PMID- 3025223 TI - Cell surface proteoglycan associates with the cytoskeleton at the basolateral cell surface of mouse mammary epithelial cells. AB - The cell surface proteoglycan on normal murine mammary gland mouse mammary epithelial cells consists of an ectodomain bearing heparan and chondroitin sulfate chains and a lipophilic domain that is presumed to be intercalated into the plasma membrane. Because the ectodomain binds to matrix components produced by stromal cells with specificity and high affinity, we have proposed that the cell surface proteoglycan is a matrix receptor that binds epithelial cells to their underlying basement membrane. We now show that the proteoglycan surrounds cells grown in subconfluent or newly confluent monolayers, but becomes restricted to the basolateral surface of cells that have been confluent for a week or more; Triton X-100 extraction distinguishes three fractions of cell surface proteoglycan: a fraction released by detergent and presumed to be free in the membrane, a fraction bound via a salt-labile linkage, and a nonextractable fraction; the latter two fractions co-localize with actin filament bundles at the basal cell surface; and when proteoglycans at the apical cell surface are cross linked by antibodies, they initially assimilate into detergent-resistant, immobile clusters that are subsequently aggregated by the cytoskeleton. These findings suggest that the proteoglycan, initially present on the entire surface and free in the plane of the membrane, becomes sequestered at the basolateral cell surface and bound to the actin-rich cytoskeleton as the cells become polarized in vitro. Binding of matrix components may cross-link proteoglycans at the basal cell surface and cause them to associate with the actin cytoskeleton, providing a mechanism by which the cell surface proteoglycan acts as a matrix receptor to stabilize the morphology of epithelial sheets. PMID- 3025224 TI - Isolation and characterization of the gene for myosin light chain two of Drosophila melanogaster. AB - A recombinant lambda-phage DNA clone containing Drosophila melanogaster sequences encoding the gene for myosin light chain (MLC) two has been isolated from a library of randomly sheared DNA. The Drosophila MLC2 gene is located in region 99E1-3 on the right arm of chromosome 3, several bands removed from the site reported for the other myosin light chain gene at 98B. The MLC2 sequence at 99E1 3 appears to encode all of the isoforms of Drosophila MLC2. The polypeptide encoded at 99E was identified as MLC2 by the following criteria: the in vitro translation product is identical in size to MLC2 isolated from Drosophila muscle, and on two-dimensional gels the in vitro translation product can be separated into two or more peptides that co-migrate with isoforms of larval and thoracic MLC2. RNA encoding the polypeptide was detected in embryos only after the onset of muscle differentiation and was also abundant in adult thoracic muscle. The nucleotide sequence of cDNA generated from late embryonic RNA would be translated to yield a protein sequence with multiple regions of homology to vertebrate MLC2. (There are shorter regions of homology to vertebrate MLC1). Like a number of vertebrate muscle proteins, Drosophila MLC2 has an acetylated amino-terminus. PMID- 3025226 TI - Peripheral nerve surgery. PMID- 3025225 TI - Signal recognition particle causes a transient arrest in the biosynthesis of prepromelittin and mediates its translocation across mammalian endoplasmic reticulum. AB - The translocation of prepromelittin (pPM) across mammalian endoplasmic reticulum was studied in both wheat germ and reticulocyte lysate. In the wheat germ system, signal recognition particle (SRP) caused a transient arrest in the synthesis of pPM. This was indicated by a slowdown in the rate of synthesis of pPM in the presence of SRP. The arrest was specific, dependent on the concentration of SRP, and more effective at early incubation time. In a tightly synchronized translation system, SRP had no apparent effect on the elongation of pPM, indicating that the effect of SRP on pPM chain synthesis might be at the final stages of chain elongation and release from the ribosome. This was reflected in a transient accumulation of pPM as peptidyl tRNA. Because pPM is composed of only 70 amino acids, arrest by SRP may be very close to chain termination. Arrest at this stage of chain synthesis seems to be unstable and the nascent chain gets terminated and released from the ribosome after a transient delay. The translocation of pPM was shown to be dependent on both SRP and docking protein. The difference in the translocation efficiency of pPM in reticulocyte and wheat germ lysates may reflect a difference in the targeting process in the two systems. PMID- 3025227 TI - Transmission and scanning electron microscopy of peripheral nerves. AB - Electron microscopy of peripheral nerve tissue by SEM and TEM facilitates a better understanding of the complex nerve architecture described by early light microscopists. Details of axonal morphology and the Schwann cell/myelin sheath complex in healthy and diseased nerves have been described in the literature. The techniques of tissue harvesting and processing and the observed appearance of peripheral nerves by electron microscopy should be useful to clinicians and researchers involved in peripheral nerve surgery and research. PMID- 3025228 TI - The significance of longitudinal excursion in peripheral nerves. AB - It has been shown that nerves do, in fact, have a longitudinal excursion. This excursion is accentuated during adjacent joint motion. It is important for the surgeon treating nerve injuries, compression lesions, and adherent nerves to realize that the restoration of the longitudinal excursion of the peripheral nerves must be accomplished in order to effectively treat the inciting lesion. PMID- 3025229 TI - Comparison of the relative importance of tyrosine-specific vinculin phosphorylation and the loss of surface-associated fibronectin in the morphology of cells transformed by Rous sarcoma virus. AB - We have investigated the relative importance of tyrosine-specific phosphorylation of vinculin and the loss of surface-associated fibronectin in the maintenance of the rounded morphology characteristic of chick embryo fibroblasts (CEF) transformed by Rous sarcoma virus (RSV). To address this question we have examined the interaction of CEF and RSV-CEF in vitro with exogenously added fibronectin in both 3-day culture experiments and short-term, 3-h spreading experiments. We report that the addition of human plasma fibronectin to cultures of RSV-CEF results in the restoration of a near-normal morphology, as has been described previously, with the added fibronectin incorporated into an extracellular matrix. However, the phosphotyrosine content of vinculin in these cells was unchanged from that of control, untreated RSV-CEF despite the change in morphology. In short-term spreading experiments RSV-CEF were unable to adopt a fully spread morphology on fibronectin substrates, with defects in the formation of adhesion plaques and microfilament bundles compared with untransformed CEF. pp60v-src was present in the newly formed adhesion plaques of RSV-CEF spreading on fibronectin substrates. The relevance of these results to the maintenance of the transformed phenotype is discussed. PMID- 3025230 TI - Contraction of collagen lattice by peritubular cells from rat testis. AB - Peritubular cells from 15- and 25-day-old rat testis trapped in collagen lattices caused those lattices to contract. Contraction proceeded more rapidly and to a greater extent using cells from younger rats. When 36,000 cells from 15- and 25 day-old rats were trapped in 800 mm2 lattices, the areas were reduced to 28 mm2 and 170 mm2, respectively, within 24 h. The cells from older rats were less effective at contracting the lattice than cells from younger rats. Cytochalasin B (5 micrograms ml-1) inhibited lattice contraction and caused disruption of actin filaments as seen by fluorescent staining with Rh-phalloidin. Cholera toxin (10 micrograms ml-1), and 1 mM-dibutyryl cAMP inhibited lattice contraction, as did 10 microM-trifluoperazine, commonly an inhibitor of calmodulin. The total intracellular concentration of cAMP was greater in peritubular cells from 25-day old rats than in those from 15-day-old rats: 427 +/- 34 and 120 +/- 16 pmol mg-1 cell protein, respectively. When peritubular cells in monolayer were permeabilized with glycerol, the addition of ATP caused the cells to contract. Cell contraction was greater in cells from 15-day-old rats than 25-day-old rats. When cells were grown on silicone rubber, they caused that surface to wrinkle. Peritubular cells from 15-day-old rats caused the onset of wrinkling at 4 h. At the same time, no wrinkling was observed with cells from 25-day-old rats. Studies of lattice contraction and cell contraction were also made using cells from 20 day-old rats. In each case, contraction was intermediate between that of cells from 15-and 25-day-old rats. The possibility exists that lattice contraction, cell contraction and wrinkling of silicone film result from a mechanism of actin filament sliding, generated by myosin ATPase activity, and is inhibited by cAMP. The reduced rate of contraction in cells from 25-day-old rats may be related to their higher intracellular levels of cAMP. Evidence exists to show that cAMP blocks myosin ATPase activity by inhibiting the phosphorylation of its regulatory peptide, myosin light chain. PMID- 3025231 TI - Inositol 1,4,5-trisphosphate and calcium stimulate actin polymerization in Dictyostelium discoideum. AB - The effect of chemoattractants such as cyclic AMP and folate on amoebae of the cellular slime mould Dictyostelium discoideum is to cause a series of rapid intracellular responses. One of the most rapid of these responses is the polymerization of actin associated with the cytoskeleton, an event correlated with pseudopodium formation, which occurs within 3-5 s of chemotactic receptor stimulation. We report that this response can be mimicked by addition of 5 microM inositol 1,4,5-triphosphate (IP3) or by addition of 100 microM-Ca2+ to saponin permeabilized amoebae. The data suggest that cytoskeletal actin polymerization occurs in normal cells as a result of IP3 formation in response to cell surface receptor stimulation and the consequent release of Ca2+ from internal stores. PMID- 3025232 TI - Direct cell-to-cell transmission of vesicular stomatitis virus. AB - Vesicular stomatitis virus (VSV) infection of kidney-derived, LLC-PK1 epithelial cells resulted in the budding of new viral particles into the basolateral space of the cultures. In lateral regions where cells were in close apposition, the majority of assembling viral particles in the process of budding from the producing cell had their apex already engaged in clathrin-coated pits of the neighbouring cell surface. These observations suggest that the viral envelope plasma membrane interaction triggers the focal formation of clathrin-coated pits; they also show how VSV infection could spread throughout a tissue with only minimal exposure to a host's extracellular environment. PMID- 3025233 TI - Ascorbate induced changes in glycosaminoglycan synthesis and distribution of normal and SV40-transformed fibroblasts. AB - The effect of ascorbate on the glycosaminoglycans synthesized by normal and simian virus 40(SV40)-transformed human skin fibroblasts was examined. Cells were incubated in the presence or absence of ascorbate, and radiolabelled with [3H]glucosamine and [35S]sulphate for 48 h, 3 days after reaching confluence. Glycosaminoglycans were analysed in the medium, a collagenase extract, and in the trypsin/cell-associated fraction. Hyaluronic acid was the main 3H-labelled glycosaminoglycan in all but the collagenase extracts, and showed a large decrease in normal fibroblast cultures, but a significant increase in SV40 transformed fibroblast cultures following feeding with ascorbate. Incorporation of [3H]glucosamine into sulphated glycosaminoglycans was reduced in normal fibroblast cultures but increased slightly in SV40-transformed cultures following ascorbate supplementation. [35S]sulphate incorporation remained essentially unaltered in both cell cultures. Ascorbate stimulated the deposition of glycosaminoglycans into the insoluble matrix of normal fibroblasts while reducing the deposition in SV40-transformed fibroblast cultures. The observed changes may in part be related to ascorbate-induced deposition of collagen in normal fibroblast cultures and the inability of the transformed fibroblast cells to deposit an extensive extracellular matrix, in addition to possible changes in the specific activity of the UDP-N-acetyl-[3H]hexosamine pool. PMID- 3025234 TI - Comparison of single-photon emission computed tomography with [123I]iodoamphetamine and xenon-enhanced computed tomography for assessing regional cerebral blood flow. AB - Regional CBF (rCBF) images obtained from xenon-enhanced computed tomography (XeCT) and single-photon emission computed tomography (SPECT) with N-isopropyl-p [123I]iodoamphetamine (IMP) done with a rotating gamma-camera were compared in nine patients. Both XeCT and SPECT/IMP demonstrated flow abnormalities at all sites of infarction identified by CT, while detecting reduced rCBF in areas normal by CT in eight of the nine patients. All areas that were abnormal on XeCT were abnormal on the comparable SPECT/IMP images. The major advantages of XeCT are its greater resolution and potential for noninvasive quantitation of rCBF, while the major advantage of SPECT/IMP is its visualization of the entire brain on transverse, coronal, and sagittal sections. PMID- 3025235 TI - 'Sick with the flu' for five years. PMID- 3025236 TI - Rapid purification of glucokinase and glycerokinase from Bacillus stearothermophilus by hydrophobic interaction chromatography. PMID- 3025238 TI - An ion-exchange capture technique for routine identification of faecal viruses by electron microscopy. AB - Faecal specimens from 520 patients with non-bacterial, gastroenteritis were examined by electron microscopy using four methods. These were (1) a direct dip method, (2) low-speed centrifugation, (3) ultracentrifugation and (4) a calcium phosphate method. The calcium phosphate method combined with low-speed centrifugation (750 X g, 2,100 X g) was considered overall best. The calcium phosphate method makes it possible to handle a large number of faecal specimens by saving considerable time and labour. PMID- 3025237 TI - The causes of false-positives encountered during the screening of old-world primates for antibodies to human and simian retroviruses by ELISA. AB - Sera from 526 old-world primates representing 50 different species were screened by ELISA for antibodies to human T-lymphotropic viruses I and III, and simian retrovirus type 1 (SRV-1). About one-fourth of the sera were positive by ELISA. There was a tendency, however, for the same sera to be positive for all three human and simian retroviruses. Only about one in five of the ELISA antibody positive sera were confirmed to be positive by Western blotting. False-positive ELISA antibody tests were particularly common among sera from mandrills, crab eating macaques, lion-tailed macaques, African green monkeys, and DeBrazza's and moustached guenons. Sera that were falsely positive in ELISA antibody tests to the three human and simian retroviruses were found to contain antibodies that reacted at comparable intensity with feline leukemia, infectious peritonitis and panleukopenia viruses. The false anti-viral activity of these sera was found to be due to antibodies that reacted with non-viral proteins that were copurified with all five virus preparations. These proteins were present in normal cat and human cells used to grow the various viruses and in gelatin. The implications of nonspecific cell-protein antibodies in primate sera were discussed in the light of this and previous seroepidemiologic studies of man and old-world monkeys. PMID- 3025239 TI - An improved ELISA method, using a streptavidin-biotin complex, for detecting Marek's disease virus antigens in feather-tips of infected chickens. AB - To improve sensitivity in the detection of Marek's disease virus (MDV) antigens in extracts of feather tips from infected chickens, we added a preformed streptavidin-biotin complex to the standard enzyme-linked immunosorbent assay (ELISA). Rabbit anti-chicken IgG-alkaline phosphatase that is used in the standard ELISA as the conjugate was replaced by a biotinylated rabbit anti chicken IgG plus the streptavidin-biotin peroxidase complex (ABC) system. The ABC ELISA system was correlated to the standard ELISA. There was increased sensitivity in the detection of MDV antigens present at low concentrations, while at the higher concentrations detection was similar to that in the standard ELISA. Both ELISA systems had the same increased sensitivity when compared with that of the agar gel precipitation (AGP) test. PMID- 3025240 TI - Rapid method for the identification and screening of herpesviruses by DNA fingerprinting combined with blot hybridization. AB - Rapid characterization of herpesvirus isolates exemplified by equine herpesviruses is described. Total DNAs were isolated from virus infected small scale cell cultures. The DNA fragments obtained after restriction enzyme digestion were separated on agarose gels, transferred and immobilized on filter membranes. A radioactively labelled probe derived from the purified DNA of an EHV 1 reference strain was used for hybridization in order to detect the restriction fragments of different EHV-1 field isolates. This method allows the typing of many isolates within a short period of time. PMID- 3025242 TI - A microneutralization test for the identification of enterovirus isolates. AB - Two-hundred and thirty-four enterovirus isolates were identified using a recently modified, microneutralization procedure. Neutralization tests were performed in 96 well plastic 'V' plates, using 20 microliter quantities of antisera and virus, then inoculated onto monolayers of buffalo green monkey kidney cells, growing in 48 well tissue culture plates. Utilization of this microneutralization procedure resulted in considerable savings of time, material, and particularly, neutralizing antisera. PMID- 3025241 TI - An improved in situ DNA hybridization protocol for detection of human papillomavirus (HPV) DNA sequences in paraffin-embedded biopsies. AB - In situ DNA hybridization is becoming rapidly an important technique for detection and typing of human papillomaviruses (HPV) in epithelial lesions, some of which (those due to HPV 16, 18 and 31) seem to possess an increased risk for progression into an invasive squamous cell carcinoma. An improved in situ DNA hybridization technique (Technique II) was described, and the results obtained in a series of cervical and penile HPV lesions were compared with those given by the in situ hybridization technique (Technique I) previously used in our laboratory. Special emphasis was made to increase the sensitivity with three basic alterations of the hybridization protocol; omission of the 0.2 N HCl wash, use of increased proteinase K concentration (from 50 micrograms/ml to 1 mg/ml), and elevated denaturation temperature (obtained by a heating block instead of an incubator). Poly-D-lysine as a slide-coating medium was replaced by Kodak Photo Flo 200 to improve the attachment of the tissue sections on the slides. Identical HPV DNA types were discovered by the two hybridization techniques. The attachment of the tissue sections was equal on the slides coated with either poly-D-lysine or Kodak Photo-Flo 200, and the latter did not interfere with the sensitivity of in situ hybridization. The hybridization signals for HPV DNA were weak or moderate in 15/16 lesions with Technique I, but intense in 10/16 lesions with Technique II (P less than 0.001). Furthermore, the resolution of Technique II seemed to be superior to that of Technique I, while being capable of disclosing HPV DNA in the intermediate cell layers (P less than 0.001) and in basal/parabasal cell layers (P less than 0.025) of both the cervical and penile lesions. The discovery of HPV DNA in the parabasal cells provides important clues to the understanding of the biology of HPV infection in the squamous epithelium, and makes this improved in situ DNA hybridization technique invaluable in assessing the lesions, where low copy numbers of HPV are to be expected. PMID- 3025243 TI - Use of [32P]RNA probes for the dot-hybridization detection of potato spindle tuber viroid. AB - A dot-hybridization assay using 32P-labelled RNA probes (+RNA and cRNA) transcribed from potato spindle tuber viroid (PSTV) cDNA was described. A complete cDNA copy of PSTV, originally cloned in pBR 322 (pAV 401) was subcloned in the BamHI site of a 'Riboprobe' cloning vectors pSP 64 and pSP 65 in opposite orientations. The reconstructed plasmids were designated pDX 1 and pDX 4, respectively. Transcription of pDX 1 and pDX 4 plasmids by SP6 RNA polymerase resulted in the generation of PSTV-specific RNA (+RNA) and PSTV complementary RNA (cRNA), respectively. The cRNA probe was much more sensitive than the +RNA probe and the nick-translated cDNA probe from the plasmid pAV 401 for the detection of PSTV in clarified plant sap. As little as 1.4 pg of purified PSTV mixed in clarified sap from uninoculated tomato leaves has been detected using cRNA probe. A relatively simple procedure using cetyltrimethyl ammonium bromide (CTAB) as nucleic acid precipitant and an enrichment step for the purification of PSTV was described. PMID- 3025244 TI - Two enzyme immunoassays for the detection of antibody to rodent coronaviruses. AB - Two enzyme immunoassays for detection of antibody to rodent coronaviruses were compared. Mouse hepatitis virus (MHV), strain JHM, antigen was in the form of formalin-fixed, infected 17 C1 1 cells. This antigen detected antibody to the homologous strain of MHV as well as to two heterologous MHV strains and a serologically related rat coronavirus, sialodacryodenititis virus. Antibody titers in assays using horseradish peroxidase (HRP)-conjugated or ureiase conjugated anti-mouse IgG were substantially higher than in an indirect immunofluorescence assay. The ureiase assay was somewhat more sensitive than the HRP assay. MHV-JHM antigen was stable under a variety of storage conditions for at least two months. PMID- 3025245 TI - Diminished aldosterone responses to angiotensin II and adrenocorticotropin in hypocalcemic subjects: restoration of responsiveness with normocalcemia. AB - ACTH-, angiotensin II (AII)-, and K+-mediated aldosterone responses in vitro are dependent on extracellular and intracellular Ca concentrations. This study examined in vivo the relationship of changes in ambient serum calcium (serum Ca) to ACTH- and AII-mediated aldosterone release in hypoparathyroid subjects. Plasma aldosterone (PA) responses to graded dose infusions of ACTH and AII were examined in hypoparathyroid (HypoPTH) patients before (n = 8) and after correction of hypocalcemia (n = 6) and compared to responses in 20 normotensive normocalcemic subjects. ACTH and AII were infused for 90 min at rates increasing from 12.5 to 50 mIU/30 min and 0.5 to 2.0 ng/kg X min, respectively. Pretreatment mean serum Ca was 6.8 +/- 0.2 (+/- SEM) mg/dl, and it rose to 9.3 +/- 0.2 mg/dl after 3-8 weeks of vitamin D administration. In the untreated HypoPTH patients, basal mean PA (5.4 +/- 1.3 ng/dl) was lower (P less than 0.01) than in the normal subjects (10.6 +/- 0.6 ng/dl) or treated HypoPTH patients (9.5 +/- 1.8 ng/dl). There was a marked reduction in PA responses to ACTH at all doses in the untreated HypoPTH patients compared to the normal subjects. With normalization of serum Ca in four patients, the mean peak PA response to ACTH (25.1 +/- 6.0 ng/dl) was not significantly different from normal (28.9 +/- 1.7 ng/dl). During graded dose AII infusion in five untreated HypoPTH patients, mean PA levels increased from 6.9 +/ 1.2 to 11.6 +/- 2.2 ng/dl; when the serum Ca was normal, the corresponding values were 8.7 +/- 1.8 and 20.2 +/- 3.61 ng/dl. There was a positive correlation (r = 0.475; P less than 0.05) between basal PA and serum Ca levels. In addition, maximum changes in mean arterial pressure in response to AII infusions were significantly greater after correction of hypocalcemia. These observations indicate that in HypoPTH patients, extracellular Ca concentrations can influence humoral aldosterone response to ACTH and AII and pressor responses to AII. PMID- 3025246 TI - The relationship of saline-induced changes in vasopressin secretion to basal and corticotropin-releasing hormone-stimulated adrenocorticotropin and cortisol secretion in man. AB - To investigate the effect of endogenous arginine vasopressin (AVP) on ACTH secretion, normal subjects were given infusions of either hypertonic saline (HS) or isotonic saline (NS) combined with human corticotropin-releasing hormone (CRH) or placebo. Basal plasma AVP was 2.3 +/- 0.3 (+/- SE) pg/ml, did not change with NS treatment, and rose to 5.4 +/- 0.6 pg/ml during HS infusion (P less than 0.01). Both basal and CRH-stimulated plasma ACTH and cortisol concentrations increased during HS infusion. Peak plasma ACTH and cortisol levels were 11.4 +/- 1.5 pg/ml and 8.6 +/- 0.8 micrograms/dl, respectively, during the HS (plus placebo) infusion. During the NS (plus placebo) infusion, plasma ACTH and cortisol gradually declined to 6.8 +/- 0.5 pg/ml and 2.6 +/- 0.4 micrograms/dl. The timing of the rise in ACTH during the HS infusion paralleled the rise in AVP. When an iv dose of 1 microgram/kg CRH was administered during the saline infusions, peak plasma ACTH and cortisol levels were 27.7 +/- 6.3 pg/ml and 17.5 +/- 1.0 micrograms/dl, respectively, during the HS infusion and 15.6 +/- 1.7 pg/ml and 13.4 +/- 1.2 micrograms/dl during the NS infusion. When the areas under the hormone response curves were compared, CRH stimulated ACTH and cortisol secretion to a greater extent than did HS (P less than 0.05). The hormonal stimulation due to combined CRH and hypertonic saline was greater than that attributable to either factor alone (P less than 0.025), but was not different than the sum of the effects of the individual factors. These results indicate that increases in endogenous AVP produced by HS are associated with increases in both basal and CRH-stimulated ACTH and cortisol release. The effect of HS appears to be additive to but not consistently synergistic with the effect of CRH. PMID- 3025247 TI - Increased leukotriene B4 synthesis in patients with primary hyperparathyroidism is normalized after parathyroidectomy: a study comparing parathyroidectomy to thyroid adenomectomy. AB - Synthesis of leukotriene B4 (LTB4) upon stimulation of peripheral blood samples with the calcium ionophore A 23187 was studied in patients with primary hyperparathyroidism who underwent parathyroidectomy. Preoperatively, an increased LTB4 concentration of 1.76 +/- 0.19 ng/ml plasma was found, vs. 0.95 +/- 0.28 ng/ml in healthy individuals. On the fourth day after operation, the LTB4 concentration was almost normalized, reaching 1.25 +/- 0.23 ng/ml plasma. At the same time, mean serum calcium levels were reduced from 6.1 +/- 0.6 meq/liter before operation to 4.53 +/- 0.28 meq/liter after operation. In a control group, euthyroid patients with thyroid adenomas who underwent adenomectomy had normal LTB4 levels before operation (0.84 +/- 0.11 ng/ml) and did not show significant changes in LTB4-synthesizing capacity. The results indicate that synthesis of LTB4 in vivo may depend in part on factors related to serum calcium concentration or calcium metabolism. PMID- 3025248 TI - Typing of herpes simplex virus by capture biotin-streptavidin enzyme-linked immunosorbent assay and comparison with restriction endonuclease analysis and immunofluorescence method using monoclonal antibodies. AB - A sensitive enzyme-linked immunosorbent capture assay using biotin and streptavidin (capture B/SA ELISA) was developed using type-specific monoclonal antibodies for typing of herpes simplex virus. Rabbit anti-herpes simplex virus immunoglobulin G was used as the capturing antibody, and biotin-linked type-1 specific mouse monoclonal antibody or rabbit type-1- or type-2-specific polyclonal antibody served as the detecting antibody. The captured antigen was detected by an ELISA with alkaline phosphatase-conjugated streptavidin, which reacted with biotin molecules on the detector antibody. The capture B/SA ELISA was compared with other methods for efficiency and reliability in typing. Results obtained by restriction endonuclease digestion of the radiolabeled viral genome were used to determine the type (1 or 2) of clinical isolates. These results were then used as a reference for determining the accuracy of the capture B/SA ELISA, as well as that of the immunofluorescence method, both of which are easily adaptable for use in the clinical laboratory. The three methods were in perfect agreement. It was determined that both the capture B/SA ELISA and the immunofluorescence method using monoclonal antibodies provided typing results with 100% specificity and 100% sensitivity and thus were accurate and reliable. However, the ELISA was the method of choice because of its simplicity, rapidity, and use of nonradioisotopic reagents. PMID- 3025250 TI - Cultivation and characterization of human rotaviruses with "super short" RNA patterns. AB - Two group A, subgroup I, rotavirus strains possessing "super short" RNA patterns were adapted to growth in MA-104 cells. Both produced marked cytopathic effect in primary culture. Reciprocal cross-neutralization titers, polypeptide analysis, and the serum neutralizing antibody response of an infected child suggested that super short viruses are serotypically distinct from the four recognized human serotypes. PMID- 3025251 TI - Immunoglobulin M to cytomegalovirus in primary and reactivation infections in renal transplant recipients. AB - Two commercially available enzyme immunoassays and one assembled in house were used to measure immunoglobulin M (IgM) antibody to cytomegalovirus (CMV) in a total of 220 serum specimens from 104 renal transplant recipients. All assays included a step in which interfering IgG antibody was removed or complexed. Concordance of results between pairs of assays ranged from 84 to 96%. All sera from patients with recent seroconversion (primary CMV infection) had measurable anti-CMV IgM. Among those already seropositive to CMV when transplanted, 26 to 55% had IgM antibody posttransplant, depending on the assay. This was observed regardless of the CMV serologic status of the kidney donor, indicating that reactivation of endogenous CMV, as well as reinfection, can induce this antibody in transplant recipients. Four cadaver donors known to transmit CMV to eight recipients did not have measurable IgM antibody to CMV. PMID- 3025249 TI - Epidemiology of equine herpesvirus 2 (equine cytomegalovirus). AB - The epidemiology of equine herpesvirus 2 was examined by using restriction endonuclease DNA fingerprints to distinguish viruses isolated from two groups of horses. The first group consisted of three yearlings isolated from other horses but in contact with each other for 418 days, whereas the second comprised seven mares and their foals, which were sampled at monthly intervals from parturition until the foals were about 180 days old. There was a complex pattern of transmission, with 15 different viruses isolated from both groups. Four distinguishable viruses were isolated from the three yearlings by day 16 of quarantine, and by day 141 an additional two viruses were isolated. Up to five different viruses were isolated from one yearling. Although four repeat isolations of one virus from the nasal cavity of one yearling over 54 days indicated that equine herpesvirus 2 established persistent infection with constant shedding, most repeat isolations yielded distinguishable viruses. Identical viruses were isolated from the nasal cavity and leukocytes of one yearling and the nasal cavity and vagina of another, indicating that a particular equine herpesvirus 2 strain was not site specific. Although seven different viruses were isolated from the three yearlings throughout the quarantine period, two appeared to establish latent infections; one virus was not isolated until 141 days after quarantine, whereas the second was first isolated 16 days after quarantine and then for the second time, from the same horse, 402 days later. Multiple concurrent local infections were demonstrated by the isolation of two or more viruses from the same nasal swab. PMID- 3025252 TI - Kinetics of actin monomer exchange at the slow growing ends of actin filaments and their relation to the elongation of filaments shortened by gelsolin. AB - The kinetics of actin monomer exchange with the slow growing pointed ends of actin filaments have been determined by measuring rates of monomer addition to or loss from filaments with their fast-growing barbed ends blocked by the protein gelsolin. Direct measurements of filament length by electron microscopy confirmed that each gelsolin acts as a nucleus for an actin filament. The rate constants ascertained are k- = 0.03 s-1; k+ = 0.06 microM-1 s-1 at 23 degrees C and k- = 0.11 s-1; k+ = 0.09 microM-1 s-1 at 37 degrees C. They are approximately independent of pH from 7.0 to 8.0 at both temperatures. These rates are far slower than those reported on the basis of some electron microscopic studies of filaments assembled on to actin bundles. The rate constants also predict a higher critical monomer concentration for the pointed end at 37 degrees C than at room temperature, consistent with direct measurements of this quantity. The relative slowness of the monomer exchange at the pointed end suggests that actin filaments with blocked barbed ends are relatively stable. The rate of redistribution of actin monomers from filaments stabilized at their barbed ends by gelsolin-calcium complex to longer filaments was measured following removal of Ca2+, which decreases the capacity of gelsolin to nucleate filaments. The elongation occurs at a rate consistent with the measured rates of monomer exchange and is quantitatively described by Hill's model for filament elongation by random exchange of monomers from one end. PMID- 3025253 TI - A simple and rapid preparation of fully phosphorylated and fully dephosphorylated skeletal muscle myosin. Application to the preparation of a phosphorylated LC2 modified artificial isozyme. AB - Fast skeletal myosin LC2 is phosphorylated on ser-15 by a specific myosin light chain kinase (MLCK) in the presence of Ca2+ and calmodulin, and dephosphorylated by a muscle phosphate in the presence of Mg2+. Fully dephosphorylated myosin is obtained by dialysis of muscle crude extract (0.06 M NaCl, 0.01 M Tris-HCl, pH 7.5, 50 microM EGTA); fully phosphorylated myosin is obtained by addition of Ca2+ (0.2 mM), Mg2+ (10 mM) and ATP (3 mM) and 5 min incubation at 28 degrees C. The following reaction characteristics were noted. The crude extract is a very efficient phosphorylating complex and can be diluted to phosphorylate or dephosphorylate purified myosin. Phosphorylation and dephosphorylation appear monophasic, showing no evidence of negative cooperativity in this particular type of myosin and medium. Phosphorylation is 24 times slower in the presence of 0.45 M KCl, 5 mM pyrophosphate. Thiophosphorylated myosin is slowly dephosphorylated by phosphatase. At the crude myosin stage the dephosphorylation reaction is efficiently inhibited (at 0-4 degrees C) by the presence of 70 mM NaF. Myosin [(T)-LC2'] (a myosin species in which LC2 has been selectively modified by trypsin) is an interesting species refractory to phosphorylation. The myosin-[(T) LC2'] isozyme can be obtained fully phosphorylated by phosphorylation of myosin followed by limited tryptic proteolysis as described earlier. Urea-PAGE as used separates LC2, phosphoryl-LC2, LC2' and phosphoryl-LC2' effectively and in this order. Through this procedure the (de)-phosphorylating complex is ipso facto specific to the myosin species considered; the method avoids lengthy preparations of purified proteins and is easy, rapid and efficient. PMID- 3025254 TI - Analysis of Ia induction on Lewis rat astrocytes in vitro by virus particles and bacterial adjuvants. AB - Viral particles of a neurotropic murine hepatitis virus (JHM) and various substances known to have immunoregulatory effects, including bacterial lipopolysaccharide (LPS) and synthetic adjuvant peptide (muramyl dipeptide) (AP), were tested for their ability to induce Ia antigen expression on Lewis rat astrocytes in vitro. JHM virus, LPS and AP are all capable of inducing Ia molecules on astrocytes, however, in a pattern and kinetics distinct from recombinant rat gamma interferon (gamma-IFN). Whereas gamma-IFN induced Ia expression on astrocytes and all macrophages after 48 h treatment, JHM virus, LPS and AP required 4-7 days for maximal induction of Ia on astrocytes, but had little to no effect on the macrophage population. This indicates that astrocytes are uniquely reactive to components derived from infectious agents and that these components are immunoregulatory with respect to Ia expression on astrocytes. We have also attempted to determine possible mechanisms by which these agents induce astrocyte Ia and show that phorbol myristate acetate and Ca2+ ionophore A23187 have similar effects. These findings suggest that infectious agents may directly stimulate antigen presenting functions of astrocytes in the brain through gamma IFN-independent mechanisms. PMID- 3025256 TI - Endotoxin enhances tissue factor and suppresses thrombomodulin expression of human vascular endothelium in vitro. AB - Endotoxemia is frequently associated clinically with disseminated intravascular coagulation (DIC); however, the mechanism of endotoxin action in vivo is unclear. Modulation of tissue factor (TF) and thrombomodulin (TM) expression on the endothelial surface may be relevant pathophysiologic mechanisms. Stimulation of human umbilical vein endothelial cells with endotoxin (1 microgram/ml) increased surface TF activity from 1.52 +/- 0.84 to 11.89 +/- 8.12 mU/ml-10(6) cells at 6 h (n = 11) which returned to baseline by 24 h. Repeated stimulation at 24 h resulted in renewed TF expression. Endotoxin (1 microgram/ml) also caused a decrease in TM expression to 55.0 +/- 6.4% of control levels at 24 h (n = 10) that remained depressed at 48 h. Both effects were dose and serum dependent. A temporary rise in TF expression accompanied by a sustained fall in TM expression comprise a shift in the hemostatic properties of the endothelium that would favor intravascular coagulation and may contribute to the pathogenesis of DIC in gram negative septicemia. PMID- 3025255 TI - Circulating and tissue angiotensin systems. PMID- 3025257 TI - Sialic acid glycoproteins inhibit in vitro and in vivo replication of rotaviruses. AB - We investigated the interactions of rotaviruses with glycoproteins and cells that support rotaviral replication. We found that a wide range of naturally occurring glycoproteins, including ovalbumins and ovomucoids from chicken and turkey eggs, and mucin derived from bovine submaxillary glands, inhibit the replication of rotaviruses in MA-104 cells. Our studies further indicated that the glycoproteins bind directly to rotaviruses and that virus-glycoprotein binding is dependent largely upon interactions with sialic acid oligosaccharides. We found that accessible sialic acid oligosaccharides are required for efficient rotavirus infection of MA-104 cells, thus demonstrating that sialic acid oligosaccharides play an important role in the interactions of rotaviruses with both glycoproteins and cells that support rotaviral replication. Bovine submaxillary mucin and chicken ovoinhibitor can also prevent the shedding of rotavirus antigen and the development of rotavirus gastroenteritis in a mouse model of rotavirus infection. Our findings document that a range of glycoproteins inhibit the in vivo and in vitro replication of rotaviruses and suggest that the alteration in the quantity or chemical composition of intestinal glycoproteins is a potential means for the modulation of enteric infections. PMID- 3025258 TI - Effects of vitamin A deficiency and repletion on rat insulin secretion in vivo and in vitro from isolated islets. AB - We studied the effects of vitamin A deficiency and repletion on rat insulin release and islet cellular retinol binding protein (CRBP) and cellular retinoic acid binding protein (CRABP). Biphasic insulin release from vitamin A-deficient perifused islets was markedly impaired. Release remained impaired with retinoic acid (RA) repletion, 2 micrograms/g diet compared to release from islets of rats repleted with retinol in the form of retinyl palmitate, 4 micrograms/g diet. Release normalized with RA, 8 micrograms/g diet. Vitamin A deficiency did not affect islet insulin content, cell size, number or structure. In vivo, vitamin A deficient rats had impaired glucose-induced acute insulin release and glucose intolerance, which improved with repletion. Normal islets had greater concentrations of CRBP than CRABP; vitamin A deficiency reduced CRBP but not CRABP levels. We conclude retinol is required for normal insulin secretion. Retinoic acid may substitute for retinol in this function. PMID- 3025259 TI - Studies on the mechanism of omega-hydroxylation of platelet 12 hydroxyeicosatetraenoic acid (12-HETE) by unstimulated neutrophils. AB - Stimulated platelets, in the presence or absence of aspirin, synthesize significant quantities of 12-hydroxyeicosatetraenoic acid (12-HETE), which is chemotactic and chemokinetic, and enhances mononuclear cell procoagulant activity. During a cell-cell interaction between stimulated platelets and unstimulated neutrophils, platelet 12-HETE is metabolized to 12,20 dihydroxyeicosatetraenoic acid (12,20-DiHETE) by neutrophils. Characteristics of the enzyme system in unstimulated neutrophils responsible for this omega hydroxylation were investigated. A broad range of cytochrome P-450 inhibitors, as well as leukotriene B4, blocked formation of 12,20-DiHETE. Owing largely to released proteases, neutrophil homogenization abolished activity. Pretreatment with diisopropylfluorophosphate preserved activity in neutrophil homogenates. omega-Hydroxylation of 12-HETE was confined solely to the microsomal fraction. Specific activity increased 6.6-fold compared with neutrophil sonicates. The electron donor NADPH was a required cofactor. These results indicate that the enzyme in unstimulated human neutrophils, which metabolizes 12-HETE from stimulated platelets to 12,20-DiHETE in this cell-cell interaction, is a cytochrome P-450 monooxygenase. PMID- 3025260 TI - Stimulation of inositol trisphosphate and diacylglycerol production in renal tubular cells by parathyroid hormone. AB - Parathyroid hormone (PTH) produced a dose-dependent immediate stimulation of inositol triphosphate and diacylglycerol production in the opossum kidney cell line, primary culture proximal tubular cells, and basolateral membranes from canine proximal tubular segments. The increase in inositol triphosphate production was accompanied by a minor increase in inositol phosphate and no significant increase in inositol bisphosphate production. Associated with the changes in inositol triphosphate and diacylglycerol, there was an immediate hydrolysis of phosphatidylinositol 4'5-bisphosphate. The effect on phospholipid hydrolysis was followed by stimulation of phosphorylation of phosphatidylinositol 4' monophosphate and phosphatidylinositol. PTH produced a sudden increase in cytoplasmic Ca2+ in opossum kidney cells that persisted for approximately 1 min. Inositol triphosphate transiently increased cytoplasmic Ca2+ in saponin-treated opossum kidney and primary culture proximal tubule cells. The effects of PTH were not mimicked by cyclic nucleotides. In fact, cyclic AMP appeared to diminish inositol triphosphate production. These results demonstrate that PTH may activate renal tubular epithelial cells by the production of inositol triphosphate and diacylglycerol. PMID- 3025261 TI - Degradation of human glomerular basement membrane by stimulated neutrophils. Activation of a metalloproteinase(s) by reactive oxygen metabolites. AB - We examined the role of reactive oxygen metabolites in the degradation of human glomerular basement membrane (GBM) by stimulated human neutrophils. Neutrophils stimulated with phorbol myristate acetate (PMA) caused a significant degradation of GBM over 3 h resulting in 11.4 +/- 0.9% (SEM), n = 11 release of hydroxyproline compared with 0.3 +/- 0.09%, n = 11 release by unstimulated neutrophils. Superoxide dismutase, a scavenger of superoxide, did not inhibit the GBM degradation, whereas catalase, a scavenger of hydrogen peroxide, caused a marked inhibition (-60 +/- 7%, n = 4, P less than 0.001) of hydroxyproline release. Neither alpha-1 proteinase inhibitor, an inhibitor of elastase, nor soya bean trypsin inhibitor, an inhibitor of cathepsin G, caused any significant inhibition of GBM degradation. GBM degradation by cell-free supernatants obtained from stimulated neutrophils was markedly impaired in the presence of metal chelators EDTA (-72 +/- 7, n = 6, P less than 0.001) and 1,10,phenanthroline (-85 +/- 5%, n = 3, P less than 0.001). Considering these results, we postulated that reactive oxygen metabolites generated by the stimulated neutrophils activate a latent GBM degrading metalloproteinase(s). GBM degradation by supernatants obtained from incubations with catalase, azide, an inhibitor of myeloperoxidase, and methionine and taurine, scavengers of hypochlorous acid, was markedly reduced. Our data thus indicate that degradation of the GBM by PMA-stimulated neutrophils is due to activation of a latent metalloproteinase by hypochlorous acid or a similar oxidant generated by the myeloperoxidase-hydrogen peroxide halide system. PMID- 3025262 TI - Low sodium diet corrects the defect in lymphocyte beta-adrenergic responsiveness in hypertensive subjects. AB - To determine the role of dietary sodium intake in the reduction in beta adrenergic sensitivity in hypertension, lymphocyte beta-receptors from 8 borderline hypertensive and 16 normotensive subjects were studied after 5 d on a high sodium diet (400 meq/d) and also following a low sodium diet (10 meq/d). During the high sodium diet, lymphocyte beta-receptor-stimulated adenylate cyclase activity, expressed as the relative increase over basal levels stimulated by the beta-agonist isoproterenol, was significantly (P less than 0.025) decreased in hypertensive (24 +/- 5%, mean +/- SE) compared with normotensive (42 +/- 4%) subjects. Neither beta-receptor density nor the proportion of nonsequestered beta-receptors differed between groups. A low sodium diet significantly increased beta-receptor-stimulated adenylate cyclase activity in hypertensives (low sodium, 51 +/- 7%; high sodium, 24 +/- 5%, P less than 0.025) to a level not different than that of normotensives (46 +/- 5%). Thus, reduced lymphocyte beta-receptor responsiveness in hypertensive subjects is not due to beta-receptor sequestration and is corrected on a low sodium diet. Dietary sodium may be an important factor in the beta-receptor defect in early hypertension. PMID- 3025263 TI - Suppressor T cell clones from patients with acute Epstein-Barr virus-induced infectious mononucleosis. AB - Suppression and/or cytotoxicity are believed to play an important role in the defense against Epstein-Barr virus (EBV) infection. To analyze the role of suppressor T cells in relation to EBV, we sought to clone and study these T cells. Analysis of 152 T cell clones derived from the peripheral blood of two patients with acute EBV-induced infectious mononucleosis (IM) yielded 11 highly suppressive clones that had no cytotoxic activity for the natural killer sensitive K562 cell line, an autologous EBV-infected cell line, or an allogeneic EBV-infected B cell line. Four of six suppressor T cell clones also profoundly inhibited EBV-induced immunoglobulin production, and five of five clones delayed the outgrowth of immortalized cells. These results indicate that during acute IM, suppressor T cells capable of inhibiting B cell activation in the absence of cytotoxicity can be identified, and may play a key role in the control of EBV infection. PMID- 3025264 TI - Malignant fibrous histiocytoma of the orbit. AB - A recent review of orbital tumors at the Armed Forces Institute of Pathology has shown that fibrous histiocytoma is the most common primary mesenchymal tumor of the orbit in adults. This surprising statistic is explained by the fact that the histologic classification of mesenchymal neoplasms has been revised extensively during the last two decades to include previously distinct tumors of fibrous tissue under the common histopathologic diagnosis of fibrous histiocytoma. We present the computed tomographic findings of a primary malignant fibrous histiocytoma of the orbit, along with a review of the literature. PMID- 3025265 TI - Paraneoplastic tonic pupils. AB - Tonic pupils developed in two patients with malignancies outside the nervous system. Symptoms and signs of more generalized somatic and autonomic nervous system involvement were also present. Although the exact morphologic basis for autonomic dysfunction in patients with paraneoplastic neurologic deterioration is uncertain, recent studies suggest that in some cases an autoimmune mechanism is responsible and may be directed against autonomic ganglion cells. PMID- 3025266 TI - Histological and immunocytochemical study of cervical intraepithelial neoplasia (CIN) with associated HPV 6 and HPV 16 infections. AB - Histological and immunocytochemical features of cervical intraepithelial neoplasia (CIN) associated with HPV 6 and HPV 16, either singly or in combination, were studied in 48 cases. Features of HPV infection (koilocytosis, binucleation, multinucleation, giant irregular nuclei and individual cell dyskeratosis) were present in high prevalence in both HPV 6 and HPV 16 associated CIN. Abnormal mitoses seemed to be a good indicator of CIN and were present in about 50% of cases of CIN associated with either HPV 6 or HPV 16 infection. This finding provides no support for the view held by some investigators that associated HPV 16 infection can be predicted by the presence of abnormal mitoses. Expression of HPV antigen was shown in about 40% of cases with a slight, but not significantly, higher prevalence in cases of combined HPV 6 and HPV 16 infection. Conventional histology and immunocytochemistry could not distinguish CIN associated with HPV 6 from CIN associated with HPV 16 infection. PMID- 3025267 TI - Serum angiotensin converting enzyme in pneumonias. AB - Serum concentrations of angiotensin converting enzyme (ACE) were studied in pneumonias caused by different pathogens and in cases in which the aetiology could not be defined. In all aetiological groups, except in viral pneumonia, there was a significant increase in ACE during recovery (p less than 0.001). In several patients the lowest values during the acute phase of disease and the highest values during recovery were outside the reference limits. In cases with known aetiology the highest ACE values and the difference between the lowest and the highest values correlated positively with C-reactive protein concentrations at admission (p less than 0.001). The pathophysiology behind the fluctuations of the ACE concentrations is unknown. PMID- 3025268 TI - Importance of anticomplement immunofluorescence antibody titration for diagnosing varicella-zoster virus infection in Bell's palsy. AB - Anticomplement Immunofluorescence was used for antibody titration against varicella-zoster virus (VZV) in 43 patients with peripheral facial palsy. Nine of 31 patients (29%) with Bell's palsy and eight of 12 patients (75%) with Ramsey Hunt syndrome had anticomplement immunofluorescence antibody titres of greater than or equal to 1/10. On the other hand, none of 14 patients with herpes simplex virus (HSV) infection and 51 healthy adults showed anticomplement immunofluorescence antibody titres of greater than or equal to 1/10. The anticomplement immunofluorescence antibody titre in two patients with Ramsey-Hunt syndrome increased later and decreased sooner than the indirect immunofluorescence antibody titre, becoming undetectable at 66 and 104 days, respectively, after onset of the disease. There was no cross reaction between anti-VZV and anti-HSV antibodies in the patients who showed a positive antibody rise for VZV. As the acute stage of VZV infection is obscure in the patients with peripheral facial palsy without herpes the screening of anticomplement immunofluorescence antibody to VZV at titres greater than or equal to 1/10 may be useful for the diagnosis of VZV infection in patients with peripheral facial palsy. PMID- 3025269 TI - Human parvovirus associated with erythroblastopenia in iron deficiency anaemia. PMID- 3025270 TI - Localization of glutamic-acid-decarboxylase-immunoreactive axon terminals in the inferior olive of the rat, with special emphasis on anatomical relations between GABAergic synapses and dendrodendritic gap junctions. AB - Immunocytochemical and electron microscopic methods were used to examine the GABAergic innervation of the inferior olivary nucleus in adult rats. This neuronal system was visualized with an antibody against glutamic acid decarboxylase (GAD, EC 4.1.1.15), the GABA-synthesizing enzyme. A GAD-positive reaction product was encountered only in short segments of preterminal axons and in axon terminals. Their relative number per unit area of neuropil was very similar in all olivary subnuclei. Despite this homogeneity in density, obvious intraregional differences existed. Some regions were strongly immunoreactive (the "c" subgroup, the beta nucleus, and the mediolateral outgrowth of the medial accessory olive), whereas others were weakly labeled (the dorsomedial cell column and the central zones of the medial accessory and principal olives). The strongly immunoreactive areas contained the largest and most intensively labeled axon terminals. Areas of weak labeling were filled with small, weakly immunoreactive nerve terminals. Thus, variations in size and in intensity of labeling create a specific pattern of GABA innervation, revealed by an almost continuous gradient between the above-mentioned extremes. The GAD-positive axon terminals established conventional synapses with dendrites (94% of the samples) or with cell bodies (6%). The vast majority of these synapses were type II (84%) and only a small proportion formed type I synaptic contacts (16%), regardless of the nature of the postsynaptic element. Immunoreactive terminals were also involved in the complex synaptic arrangements--the glomeruli, which characterize the olivary neuropil. Within these formations, olivary neurons were electrotonically coupled through dendrodendritic gap junctions. There was a constant association between GAD positive axon terminals and small dendritic appendages linked by gap junctions. This association was revealed not only by the systematic presence of immunolabeled terminals directly apposed to the dendritic appendages but, more importantly, by the frequent presence of type II synapses straddling both elements. These synapses were in close proximity to the low-resistance pathways represented by the gap junctions. The strategic location of these GABA synapses is discussed in relation to recent findings indicating the possibility of a synaptic modulation of the electrical coupling: the release of GABA, by increasing nonjunctional membrane conductance, could shunt the coupling between olivary neurons. The functional decoupling of selected gap junctions would be responsible for the spatial organization of the olivary electrotonic coupling. PMID- 3025271 TI - Neurotensin binding sites in the forebrain and midbrain of the pigeon. AB - An autoradiographic method was used to assess the distribution of binding sites for [3H]neurotensin (NT) in the forebrain and midbrain of the pigeon. Within the telencephalon the highest levels of NT binding sites were observed within the hyperstriatum ventrale (HV). Moderate to high levels of NT binding were observed within the archistriatum, neostriatum intermedium, and hyperstriatum accessorium. These telencephalic regions and HV are thought to be comparable to portions of mammalian neocortex. Lower levels of binding sites were observed within the striatal complex including the laterally situated paleostriatum augmentatum and medially situated lobus parolfactorius. The lowest levels of NT binding sites in the telencephalon were observed within the paleostriatum primitivum (PP, considered comparable to mammalian globus pallidus), ectostriatum (comparable to layer IV of mammalian extrastriate visual cortex), field "L" (comparable to layer IV of mammalian auditory cortex), hippocampus, septum, and preoptic area. Despite considerable regional variation, the overall level of NT binding throughout the pigeon telencephalon appears to be significantly higher than that reported for mammals, particularly within pallial areas. Within the brainstem, moderate levels of NT binding sites were observed in the lateral habenular nuclei, the ventral tegmental area of Tsai, nucleus tegmentipedunculopontinus, pars compacta (comparable to the mammalian substantia nigra, pars compacta), locus coeruleus, and the nucleus subcoeruleus dorsalis. The latter four cell groups contain numerous catecholaminergic neurons. Corresponding catecholaminergic cell groups in mammalian forms also contain high levels of NT receptors. As in mammals, lower levels of NT binding were observed in most diencephalic nuclei. Somewhat higher levels of NT binding were observed within the pretectal nuclei spiriformis lateralis and spiriformis medialis. Moderate levels of NT binding sites were observed within the retinal terminal layers of the tectum (i.e., layers 1-7). Immunohistochemical experiments (Reiner and Carraway; Brain Res. 341:365-371, '85; Reiner: ARVO Abstracts: p. 185, '86) localizing NT and a related hexapeptide, LANT6, have shown that LANT6 is present in retinal ganglion cells, in cells of the paleostriatum, and in the striatotegmental and striatopretectal fiber pathways of the pigeon. Thus some features of the NT binding observed here in pigeon brain including the existence of substantial NT binding sites in the brainstem catecholamine nuclei, pretectum (nucleus spiriformis lateralis), and optic tectum may reflect the existence, at least in part, of functional receptors for LANT6.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3025272 TI - Changes in the numerical density of synaptic contacts with long-term potentiation in the hippocampal dentate gyrus. AB - Long-term potentiation (LTP) in the rat dentate gyrus is a multifaceted phenomenon, including synaptic potentiation; simultaneous synaptic depression at neighboring, unconditioned synapses; and a change in the amount of cell firing produced by a specified amount of synaptic current (see Levy and Desmond: In G. Buzsaki and C. Vanderwolf (eds): Electrical Activity of The Archicortex. Budapest: Akademiai Kiado, pp. 359-373, '85b). This study presents long-term anatomical modifications that seem related to excitatory synaptic modification. These anatomical alterations appear early and persist for at least 60 minutes following conditioning stimulation. Each animal received test pulse stimulation delivered alternately to the angular bundles before and after brief, unilateral high-frequency conditioning stimulation that is typical of many LTP paradigms. Anatomical preparation followed standard procedures. Double-blind scoring procedures quantified the number of asymmetric synapses in the dentate molecular layer. These counts were converted to the number of synapses per unit volume using stereological corrections that combined geometrically derived theory and modest serial sectioning. Multivariate analysis of variance evaluated the statistical significance of changes in synapse density. Across all three groups of animals, conditioning stimulation does not significantly change the density of synaptic contacts across the entire molecular layer. There is a trend for a decreased density of synaptic contacts in the middle molecular layer, the region activated by the conditioning stimulation. Here the density of concave spine profiles increases significantly in all three groups of animals with conditioning stimulation. This increase accompanies significant decreases in the density of nonconcave, simple and ellipsoid, spine profiles. No significant changes in the density of shaft synapses occur with LTP-inducing conditioning stimulation. These data suggest that the concave spine profiles are a correlate of LTP-inducing stimulation and may be the potentiated synapses. We hypothesize that with synaptic potentiation there occurs an interconversion of spine synapses such that some nonconcave spine profiles become concave spine profiles. Such an interconversion apparently begins shortly after the conditioning stimulation and persists for at least 60 minutes. PMID- 3025273 TI - Changes in the postsynaptic density with long-term potentiation in the dentate gyrus. AB - The present study documents alterations in the size of the postsynaptic density (PSD) of synapses formed by entorhinal afferents with granule cell dendritic spines with long-term potentiation (LTP). These changes appear early and persist for at least 60 minutes after LTP-inducing conditioning stimulation. Each animal received test and conditioning stimulation typical of LTP paradigms. Electron microscopic preparation of the dentate gyri from each animal followed conventional procedures. PSD trace lengths of identified asymmetric synaptic profiles were measured. The total PSD length for four categories of synaptic profiles was determined for each third of the molecular layer. PSD surface area per unit volume of tissue (SV) was then computed from these data. Statistical analysis of the SV data used multivariate analysis of variance. PSD surface area per synapse was also estimated. Total PSD surface area per unit volume does not change significantly throughout the entire molecular layer with LTP-inducing conditioning stimulation. However, in the activated portion of the molecular layer, total PSD surface area per unit volume tends to increase with conditioning stimulation. In the middle third of the molecular layer, total PSD surface area per unit volume associated with the concave spine profiles increases significantly while there is a statistically significant decrease in total PSD SV associated with the nonconcave spine profiles. The PSD surface area per synapse also increases markedly. Since it seems that there is an interconversion of spine synapses from nonconcave to concave with LTP (Desmond and Levy: J. Comp. Neurol. In press, '86a), these data suggest that potentiated synapses have larger responses because, in part, they have larger neurotransmitter receptive regions. PMID- 3025274 TI - Adenylate cyclase in sarcoplasmic reticulum of skeletal muscle: distribution, orientation, and regulation. AB - We found specific activity of adenylate cyclase (AC) to be as high in rat skeletal muscle sarcoplasmic reticulum (SR) as in sarcolemma (SL) (39 +/- 5 pmol/mg per min and 34 +/- 5 pmol/mg per min). Detection of AC in SR could not be due to SL contamination. Activity in SR was similar in triads (heavy SR) and longitudinal reticulum (light SR), despite virtual absence of surface membrane markers in preparations of light SR. Also, AC of SR and SL may differ biochemically. In the presence of 10(-5) M 5'-guanylylimidodiphosphate, 10(-4) M isoproterenol increased SL activity 508%, crude SR 46.4%, heavy SR 68.3%, light SR only 24.3%. SR activity was 50% higher at 0.32 micromolar Ca++ than at 1 nanomolar Ca++ (p less than 0.05); higher concentrations of Ca++ noncompetively inhibited activity (Ki 0.87 micromolar). In contrast, Ca++ monophasically inhibited SL activity. Permeabilization of SR vesicles with alamethacin indicated that AC is on the cytoplasmic surface of SR; its regulation by physiological changes in cytoplasmic Ca++ could influence SR Ca++ flux. PMID- 3025275 TI - Identification of a rat liver cAMP-dependent protein kinase, type II, which binds DNA. AB - A novel protein kinase which specifically binds single strand DNA was identified in rat liver by chromatography on double strand- and single strand- DNA cellulose. This protein kinase activity was stimulated by cAMP and was inhibited by the heat stable inhibitor, suggesting it was a cAMP-dependent protein kinase. Isoelectric focusing studies confirmed that the single strand DNA-binding protein kinase was indeed a cAMP-dependent protein kinase and had the same pI as cAMP dependent protein kinase, Type II. The DNA binding capacity of this kinase was primarily localized in the regulatory subunit. These results support the recent hypothesis that in addition to regulating enzymatic activity by phosphorylating proteins, cAMP-dependent protein kinase, Type II, may regulate mammalian gene expression through a mechanism similar to that in prokaryotes. PMID- 3025276 TI - NADP+ enhances cholera and pertussis toxin-catalyzed ADP-ribosylation of membrane proteins. AB - [32P]ADP-ribosylation of membrane proteins catalyzed by either cholera toxin or pertussis toxin was markedly enhanced by NADP+. The effect was concentration dependent; with 20 microM [32P]NAD+ as a substrate maximal enhancement was obtained at a concentration of 0.5-1.0 mM NADP+ for rabbit and guinea-pig liver membranes and 0.1 mM NADP+ for human erythrocyte membranes. NADP+ appears to act by inhibiting the degradation of NAD+ by NAD+-glycohydrolase (NADase) present in membrane preparations, probably as an alternate substrate for the enzyme. Among inhibitors tested (NADP+, isonicotinic acid hydrazide, imidazole, nicotinamide, L arginine methyl ester and HgCl2) to suppress the enzyme activity, NADP+ was the most effective and, at 10 mM, inhibited hepatic NADase activity by about 90%. The effect of NADP+ was much greater than that of other known effectors of ADP ribosylation such as Mg2+ and phosphate, or the NADase inhibitors, isonicotinic acid hydrazide and isonicotinamide. In membranes which contain substantial activities of NADase the inclusion of NADP+ in the assay system is necessary to achieve maximal ADP-ribosylation of membrane proteins. PMID- 3025278 TI - Muscarinic cholinergic receptor-induced enhancement of PGE1-stimulated cAMP accumulation in neuroblastoma X glioma cells: prevention by pertussis toxin. AB - Chronic treatment of neuroblastoma X glioma hybrid cells (NG 108-15) with the muscarinic cholinergic agonist carbachol, which acutely inhibits adenylate cyclase, resulted in a 104 +/- 10% increase in PGE1-stimulated cAMP accumulation. Pretreatment of intact cells with pertussis toxin can structurally modify the inhibitory regulatory protein, Gi, by ADP-ribosylation and thus abolish the acute inhibition by carbachol. Pretreatment of the cells with pertussis toxin also resulted in a 27 +/- 8% increase in PGE1-stimulated cAMP accumulation. In the pertussis toxin-treated cells, chronic treatment with carbachol did not further enhance the PGE1 stimulation. These results suggest that functional Gi is required for the development of muscarinic cholinergic-induced enhancement of PGE1-stimulated cAMP accumulation. PMID- 3025277 TI - cAMP metabolism in an S49 mouse lymphoma variant with diminished phosphodiesterase activity. AB - This report presents the results of detailed examinations of cAMP metabolism in B1r, an S49 lymphoma protein kinase variant with very low phosphodiesterase activity. Among the conclusions reached are: As expected from the low phosphodiesterase activity previously reported, the cAMP turnover rate was relatively slow (t1/2 was 18-23 min at 37 degrees C). Basal cAMP levels ranged from about 1% to 5% of cellular ATP (i.e., 20-100 microM or 100-500 pmol/mg protein). These were many times higher than in most S49 wild type cells. The high basal cAMP levels made measurements of turnover in the absence of a hormone possible. The turnover constant for cAMP in unstimulated cells was 0.030 +/- 0.003 min-1. This was not significantly different than the value measured during epinephrine stimulation, which was 0.035 +/- 0.004 min-1. These turnover values were used to determine precise levels of adenylate cyclase activity throughout the time course of epinephrine stimulation. Desensitization was both rapid and profound, with the level of adenylate cyclase activity falling by 70% within the first 4 minutes of stimulation. This suggested that desensitization may be a very major factor in the attenuation of catecholamine action, at least in some cell types. PMID- 3025279 TI - Physiological role of dietary fiber: a ten-year review. AB - It is accepted nowadays that dietary fiber is an important constituent of the diet. There is growing evidence that the low fiber Western diets and the low consumption of whole grain products are important factors in several common diseases of the large bowel. Cereal fiber differs from that present in vegetables and fruit. A low intake of cereal fiber has been implicated in cancer of the large bowel, diverticular disease of the colon and coronary heart disease. High fiber diets are often prescribed for diabetes. Although fiber consumption by British and American consumers has decreased over the past century, consumption of whole wheat breads and fiber-rich breakfast cereals has received new attention during the past ten years. PMID- 3025281 TI - Vagal receptors sensitive to lipids in the small intestine of the cat. AB - In anesthetized cats, the unitary activity of 53 sensory vagal neurons was recorded in nodose ganglia by means of extracellular glass microelectrodes. All the neurons had non-medullated fibres, with conduction velocities ranging from 0.8 to 1.2 m/s. Forty of these cells were stimulated by perfusion of the small intestine with lipids. Two types of receptors were identified: 21 endings were activated by glycerol and short chain lipids, and 19 endings were activated by long chain lipids. These receptors did not respond to either mechanical or osmotic stimulation. The discharge frequency generally increased with the concentration. The short latency suggested that they were located close to the enterocyte. The role of vagal intestinal receptors sensitive to lipids is discussed. Their functional characteristics along with previous experimental data suggest that they may be involved in the regulation of gastric emptying and alimentary behaviour, particularly satiety mechanisms. PMID- 3025280 TI - Beta-adrenoceptors in the rat parotid gland enlarged by salivariectomy and bulk diet. Effects of denervation. AB - The numbers of beta-adrenergic receptors and level of cyclic AMP (cAMP) of the parotid gland of adult female rats were determined 4 weeks after introduction of a regimen that induced a 2-fold increase in gland weight. This regimen consisted of ablation of the submandibular-sublingual glands and substitution of the normal chow diet with a bulk diet consisting of 50% inert cellulose and 50% ground solid chow. There was a 2.4-fold increase in number (density) of beta-adrenoceptors in the enlarged parotid gland when comparison was made with parotid glands of control rats. The beta-adrenoceptor present in the enlarged and normal glands was of the beta 1 subtype. Removal of either autonomic pathway at the time of partial salivariectomy and dietary substitution was followed by a small reduction in number of beta-adrenoceptors (4-9% with either denervation), but when both nerves were removed the reduction was 25%; in magnitude, these changes were generally similar to those observed with denervated parotid glands of chow-fed rats. The norepinephrine concentration of the enlarged gland was much less than that of normal glands (reduced 38%); sympathectomy of normal or enlarged parotid glands resulted in a marked lowering of norepinephrine concentrations (to 1-5% of control levels); parasympathectomy had no effect on norepinephrine concentration of enlarged parotid glands but caused a decrease in that of the parotid of normal size. Apparently, the number of beta-adrenoceptors depends on the degree of activity of both the parasympathetic and sympathetic nerves to the parotid.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3025282 TI - Beta-adrenergic receptors and cAMP levels of rat parotid and submandibular glands during chronic isoproterenol treatment. AB - The number of cell surface beta-adrenergic receptors and the level of cyclic AMP of the parotid and the submandibular gland were examined in rats treated for up to 10 days with twice daily injections of the beta-adrenergic agonist, isoproterenol. Receptor densities of 125 +/- 8.7 fmol/mg membrane protein for the parotid and 60.1 +/- 5.6 fmol/mg for the submandibular glands were found with [3H]dihydroalprenolol (beta-adrenergic receptor antagonist) binding of glands from control rats. No change from levels of controls was found in the number of beta-receptors of the submandibular gland with chronic isoproterenol stimulation; the parotid glands, on the other hand, showed a 22% decrease in dihydroalprenolol binding from the 4th until the 8th day of treatment. By day 10 of isoproterenol treatment the parotid gland demonstrated a shift from a population consisting of primarily beta-adrenergic receptors to one consisting of equal numbers of beta 1- and beta 2-adrenoceptors. The basal level of cAMP present in cell lysates remained unchanged in the isoproterenol-treated submandibular gland while the parotid gland showed a 30-40% decrease. Control and isoproterenol-treated animals demonstrated the same time course of cAMP accumulation after a single challenge with isoproterenol. PMID- 3025283 TI - Immunocytochemical localization of pro gamma MSH, gamma MSH, ACTH and beta endorphin/beta lipotrophin in the fetal sheep pituitary: an ontogenetic study. AB - An immunocytochemical staining technique was used to localize four fragments [pro gamma MSH, gamma MSH, ACTH and beta endorphin/beta lipotrophin (beta endorphin/beta LPH)] of the proopiomelanocortin molecule in both the adult and fetal sheep pituitary. In the adult sheep anterior pituitary each fragment was localized in cells that were darkly stained, stellate and widely distributed throughout the gland. The same cells, identified in three serial sections, stained with anti-pro gamma MSH, anti-ACTH and anti-beta endorphin/beta LPH. In the fetal sheep anterior pituitary all the proopiomelanocortin derived fragments were present at 38 days gestation. Between about 90 and 130 days of gestation both adult type proopiomelanocortin cells (small, stellate) and uniquely fetal cells (large, columnar) were present. Both adult-type and fetal proopiomelanocortin cells were identified in serial sections of the fetal anterior pituitary, stained with anti-pro gamma MSH, anti-ACTH and anti-beta endorphin/beta LPH. The adult intermediate lobe was immunoreactive with anti-pro gamma MSH and anti-beta endorphin/beta LPH but not with anti-gamma MSH or anti ACTH. The fetal intermediate lobe was immunoreactive with all four antisera from 60 days gestation. PMID- 3025285 TI - Symphalangism associated with constriction rings: syndactyly and brachytelophalangy in a black patient. AB - A 7-year-old girl presented with symptoms of constriction ring syndrome, syndactyly, and brachytelophalangy in the presence of proximal unilateral symphalangism. Physical findings revealed previous surgical correction of constriction rings of the right index and long fingers, a short right thumb with a hypoplastic distal phalanx, a shortened long finger with a stiff proximal interphalangeal joint and characteristic absence of skin creases, and incomplete syndactyly of the right first, second, and third web spaces. A brief review of symphalangism and a surgical treatment plan are presented. This is believed to be the first case report of symphalangism in a black patient with multiple congenital anomalies. PMID- 3025284 TI - Lack of anion effect on volume expansion natriuresis in the developing canine kidney. AB - The renal response to volume expansion with sodium chloride or sodium bicarbonate was studied in 15 newborn and 13 adult dogs. Proximal and distal nephron function were estimated using the technique of distal nephron blockade. Fractional sodium reabsorption was 99.0 +/- 0.3% in newborn and 96.6 +/- 0.06% in adult during the NaCl expansion (P less than 0.01) and 98.1 +/- 0.7% in the newborn and 93.2 +/- 0.7% in the adult during NaHCO3 expansion (P less than 0.001). With either anion the higher fractional sodium reabsorption in the newborn was due to reabsorption of a greater fraction of the load presented to the distal nephron segment. The percent of the distal sodium load that was reabsorbed was 98.0 +/- 0.6% in the newborn and 92.2 +/- 1.0% in the adult during NaCl expansion, and 96.1 +/- 1.3% in the newborn and 81.5 +/- 2.4% in the adult during NaHCO3 expansion. Differences in distal nephron chloride, potassium and bicarbonate reabsorption among the groups support the hypothesis that the enhanced distal sodium reabsorption in the newborn occurred largely in the ascending loop of Henle with NaCl expansion, while it occurred in the late distal and cortical collecting tubules with NaHCO3 expansion. There was no difference between the natriuretic responses to NaCl or NaHCO3 in the newborn (P greater than 0.20); however, the natriuretic response to NaCl was less than that to NaHCO3 in the adult (P less than 0.001). This suggests that the bulk of the sodium that escaped reabsorption in Henle's loop during NaHCO3 expansion was reabsorbed in the late distal tubule in the newborn, but not in the adult. PMID- 3025286 TI - Comparison of three tests for the detection of rotavirus in feces with standard ELISA. PMID- 3025287 TI - Significance of serum bilirubin level in response of hepatocellular carcinoma to doxorubicin. AB - To determine which of several clinical and laboratory features could be of significance in the response of hepatocellular carcinoma to doxorubicin we have analysed the frequency of remission in 143 patients treated with this drug. Of those features investigated including age, sex, presence and aetiology of underlying cirrhosis, duration of symptoms, performance grade and liver function tests, a normal serum bilirubin level was shown to be the only one on logistic regression analysis to predict a greater change of response (response rate = 46% with serum bilirubin less than 20 mumol/l and 10% with bilirubin greater than 20 mumol/l). The most likely explanation is that reduction in doxorubicin dosage according to the serum bilirubin level, based on the view that the risk of myelosuppression is thereby lessened, may lead to suboptimal dose administration. PMID- 3025289 TI - Immunocytochemical localization of monoamine oxidases A and B in human peripheral tissues and brain. AB - Monoamine oxidases (MAO; EC 1.4.3.4.) A and B occur in the outer mitochondrial membrane and oxidize a number of important biogenic and xenobiotic amines. Monoclonal antibodies specific for human MAO A or B and immunocytochemical techniques were used to visualize the respective enzymes in human placenta, platelets, lymphocytes, liver, brain, and a human hepatoma cell line. MAO A was observed in the syncytiotrophoblast layer of term placenta, liver, and a subset of neurons in brain, but was not observed in platelets or lymphocytes, which are known to lack type A enzyme. MAO B was observed in platelets, lymphocytes, and liver, but not in placenta, which contains little or no MAO B. MAO B was also observed in a subset of neurons in the brain that was distinct from that which contained MAO A. MAO A and MAO B were also observed in some glia. Unlike most tissues examined, liver cells appeared to contain both forms of the enzyme. These studies show that MAO A and MAO B can be specifically visualized by immunocytochemical means in a variety of human cells and tissues and can provide a graphic demonstration of the high degree of cell specificity of expression of the two forms of the enzyme. PMID- 3025288 TI - Cholestatic features in hepatitis A. AB - The clinical, biochemical and histological characteristics of 13 cases of acute hepatitis A were evaluated. In 10 biopsies moderate to severe cholestasis was seen consisting of bile thrombi, cholestatic liver cell rosettes and ductular transformation of hepatocytes as shown on keratin- and S-100 immunostaining. The periportal spotty necrosis may play a role in the pathogenesis of cholestasis in hepatitis A by creating an interruption in continuity of bile flow. In 6 cases, abnormal ductular epithelium was seen resembling the ductular lesion in septicemia. Such ductular lesion may be related to the accumulation of leucotrienes as a result of cholestasis. PMID- 3025291 TI - Localization of 125I-atrial natriuretic factor (ANF)-binding sites in rat renal medulla. A light and electron microscope autoradiographic study. AB - Using light and electron microscope autoradiography in vivo, the localization of 125I-(Arg 101-Tyr 126) atrial natriuretic factor (ANF)-binding sites was studied in the renal medulla of rats. At the light microscopic level, the autoradiographic reaction was mainly distributed in patches in the outer medulla, and followed the tubular architecture in the innermost part of the inner medulla. At the electron microscopic level, binding sites were mainly found in the outer medullary descending vasa recta and inner medullary collecting ducts. These results suggest that, in rats, the renal medulla may participate in the natriuresis and diuresis produced by ANF through vascular and tubular effects; the former by changing medullary blood flow at the level of descending vasa recta and the latter by acting on electrolyte and water transport at the level of collecting ducts. PMID- 3025290 TI - Distribution of phosphodiesterase I in normal human tissues. AB - Phosphodiesterase I (PDE I) is an exonuclease capable of hydrolyzing a variety of phosphate ester and pyrophosphate bonds. Cell fractionation and histochemical studies in animal tissues have localized PDE I in the plasma membrane of various epithelia. This suggests a role for the enzyme in active transport. Distribution of PDE I in human tissues has not previously been studied. We have produced a polyclonal antiserum to bovine intestinal PDE I and have demonstrated crossreactivity with the human intestinal enzyme. This polyclonal antiserum was used in PAP immunocytochemistry to localize immunoreactive PDE I in a variety of human tissues. Localization was prominent in the gastrointestinal tract, including the cytoplasm of gastric mucosa parietal cells, cytoplasm of surface epithelium and isolated crypt cells in small intestine, and the colonic epithelial cytoplasm and brush border. Parotid gland acinar cells and scattered ductal cells showed positive cytoplasmic staining. Acinar and scattered pancreatic islet cells contained immunoreactive PDE I, as did Kupffer cells of the liver sinusoids. Immunoreactive PDE I was found in all vascular endothelia. The epithelium of the urinary tract showed extensive immunoreactivity. This included the distal convoluted and collecting tubules of the kidney, and ureteral and bladder urothelium. In previous histochemical studies of animal tissues, no evidence of PDE I activity was noted in male or female reproductive tract. In this study, immunoreactive PDE I was localized to human Sertoli cells and to basal epithelium of the epididymis and prostate acini. Fallopian tube epithelium of female reproductive tract also demonstrated immunoreactive PDI I, as did several cell types in term placenta. Our immunocytochemical results with human tissues differ significantly from previous histochemical studies in animal tissues, principally in the genitourinary system. This may be due in part to the different detection systems employed as well as the higher sensitivity of the immunoperoxidase technique. This underscores the importance of adjunct techniques in tissue surveys. The widespread epithelial distribution of immunoreactive PDE I detected by this polyclonal antibody implies an integral role in cell function, probably in active transport. PMID- 3025292 TI - Prenatal exposure to alcohol alters the Golgi apparatus of newborn rat hepatocytes: a cytochemical study. AB - The effect of prenatal exposure to ethanol on the Golgi apparatus of newborn rat hepatocytes has been studied cytochemically using several trans-Golgi markers (thiamine pyrophosphatase, uridine diphosphatase, inosine diphosphatase, acid phosphatase, and 5'-nucleotidase) as well as a cis-side marker (osmium impregnation). The amount of cerium phosphate formed in the cytochemical reactions was roughly quantitated by stereologic methods. The Golgi apparatus of about 40% of the hepatocytes appeared disorganized after alcohol treatment, and in the other 60%, the electron density of reaction product deposits for all phosphatases investigated was decreased. 5'-Nucleotidase was completely absent in cisternae of Golgi apparatus of treated cells. In control cells impregnated with osmium tetroxide, reduced osmium compounds were observed in most Golgi cisternae and in nearby vesicles. In contrast, only small vesicles appeared positive in treated hepatocytes. These results suggest that prenatal alcohol exposure alters some Golgi functions. Thus, the decrease in nucleoside diphosphatase and 5' nucleotidase cytochemical activities after ethanol exposure strongly suggests that this treatment could affect glycosylation in the Golgi apparatus of newborn rat hepatocytes. PMID- 3025293 TI - Polymyxin B binding sites in Escherichia coli as revealed by polymyxin B-gold labeling. AB - A complex of polymyxin B, bovine serum albumin, and colloidal gold was prepared and used for the ultrastructural localization of polymyxin B binding sites on thin sections of Epon-embedded Escherichia coli cells. Gold particles were found on the outer membrane of E. coli, which is consistent with reported biochemical findings. We concluded that gold labeling with polymyxin B is useful in localizing the binding sites of polymyxin. PMID- 3025294 TI - Comparison of the intracellular pathways of transferrin recycling and vesicular stomatitis virus membrane glycoprotein exocytosis by ultrastructural double-label cytochemistry. AB - Transferrin is taken up by receptor-mediated endocytosis into intracellular vesicles and tubules, and then recycles rapidly to the plasma membrane (diacytosis). We applied double-label cytochemistry to study whether the recycling structures containing transferrin fuse with the intracellular membranous structures that deliver newly synthesized membrane glycoproteins from the ER to the plasma membrane (exocytosis) or whether they remain independent. KB and Vero cells were infected with the temperature-sensitive transport mutant 0-45 of vesicular stomatitis virus (VSV). Temperature-regulated exocytosis of membrane glycoprotein "G" occurred simultaneously with diacytosis of transferrin. The exocytic "G" protein, as detected by immunoperoxidase electron microscopy, passed through the cisternal Golgi stacks and vacuolar, tubular, vesicular, and pit-like structures of the Golgi system. A transferrin-ferritin conjugate used in ultrastructural double-label experiments was detected in diacytic vesicles and tubules that accumulated in the proximal (trans-reticular) Golgi area of the cell. The ferritin-labeled vesicles/tubules were often close to and intermixed with the VSV-"G" containing membranous structures, but in most cases at early times (15-20 min) the transferrin and VSV-"G" containing vesicular structures remained distinct. At later times (30-45 min), the two labels were occasionally found in the same structures. These results indicate that rapid recycling of endocytosed materials and exocytosis of membrane glycoproteins to the cell surface usually occur in distinct vesicles, possibly along the same general morphologic exit pathway. PMID- 3025295 TI - Simultaneous visualization of myeloperoxidase reactivity and immunogold labeling by backscattered electron imaging with the scanning electron microscope. AB - We describe a method to label circulating human granulocytes with an immunogold marker and then incubate them for demonstration of myeloperoxidase activity. The cells were observed in the backscattered electron imaging (BEI) mode of the scanning electron microscope. This permits simultaneous visualization of cell surface morphology, the immunological surface marker, and the cytochemical reactivity of each individual cell. Enhanced identification of the various cell types and more precise characterization of cell surface features in the different steps of leukocyte differentiation are expected to result from application of this technique. PMID- 3025297 TI - Effects of ouabain on blood pressure regulation in rats. AB - In order to test the hypothesis that a circulating inhibitor of the sodium potassium ATPase pump may cause a concomitant rise in blood pressure and increased sodium excretion, we studied chronic effects of continuous infusion of ouabain, an inhibitor of sodium-potassium ATPase, for up to 6 days on systolic blood pressure and urinary sodium excretion in conscious rats. We also evaluated the effect of this substance in rats with hypertension induced by chronic infusion of norepinephrine. Continuous infusion of ouabain (1.2 mg/kg per day) into the jugular vein by an osmotic minipump did not induce any changes in systolic blood pressure and urinary sodium excretion in intact rats on regular diets. Furthermore it did not cause a change in systolic blood pressure in rats drinking 1% NaCl, and in unilaterally nephrectomized rats drinking 1% NaCl, when compared with vehicle-infused animals. When the same dose of ouabain was administered simultaneously with 1.8 mg/kg per day norepinephrine infused intraperitoneally by another osmotic minipump in conscious rats, systolic blood pressure rose on day 1 to only 129.3 +/- 2.8 mmHg compared with the rist to 145.0 +/- 2.0 mmHg when norepinephrine alone was infused (P less than 0.01). The antihypertensive effect of ouabain was sustained for the entire experimental period lasting for 6 days and was not associated with any changes in urinary sodium excretion. The administration of ouabain to rats made hypertensive by a 3 day infusion of norepinephrine, returned the blood pressure to control levels, and the antihypertensive effect was sustained throughout the experimental period lasting a further 3 days and was not associated with any changes in urinary sodium excretion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3025296 TI - Prevalence of human (H1N1) influenza virus-antibody in Japanese swine. AB - A total of 571 swine sera collected at an abattoir in the city of Obihiro, Hokkaido during the period February-November 1984 were tested for antibody against human (H1N1) influenza virus strains. A high prevalence of antibody was observed for only 3 months from April to June in that year, in 81/180 sera (45.0%) to A/USSR/92/77 strain and in 50/180 sera (27.8%) to a current epidemic strain (A/Hokkaido/1/84). Some cross-reactions were observed between the A/USSR/92/77 and A/Hokkaido/1/84 antibodies (r = 0.75). Only minor relationships were noted between the A/New Jersey/8/76 (swine type H1N1) and A/USSR/92/77 (r = 0.35) or A/Hokkaido/1/84 (r = 0.51) antibodies. Absorption of sera positive for antibody to the A/Hokkaido/1/84 strain with the homologous virus strain removed all detectable antibodies, while the absorption of the sera with the A/New Jersey/8/76 strain produced incomplete absorption in one half of the sera tested. These results strongly suggest that the swine became infected with a human H1N1 virus as piglets during an epidemic of influenza which occurred in the human population during January and February 1984. PMID- 3025298 TI - Analysis of immune complexes in rheumatoid arthritis for Epstein-Barr virus antigens reveals cross-reactivity of viral capsid antigen and human IgG. AB - We recently defined the immunochemical characteristics of immune complexes (IC) isolated from synovial fluid (SF) of patients with rheumatoid arthritis with the use of Western blot analysis. In the present study, we probe for exogenous antigens in the IC by examining the specificity of antisera raised against the IC. Anti-IC antisera demonstrated strong reactivity against the viral capsid antigen (VCA) of Epstein Barr virus (EBV), which was not explained by preimmune reactivity, polyclonal B cell activation, or Fc-mediated binding in the immunofluorescence or ELISA systems used to measure antibody titers. However, comparable anti-VCA reactivity was detected in antisera raised against non rheumatoid SF. This phenomenon was not due to antigen since monoclonal anti-VCA antibody probing the IC by Western blot detected only IgG, nor to idiotype/anti idiotype interaction since normal IgG absorbed out the anti-VCA reactivity. A monoclonal anti-VCA antibody competitively inhibited the binding of anti-IgG to IgG, and Fc fragment of IgG competitively inhibited the monoclonal antibody binding to VCA. No relationship between IgG anti-VCA antibody and IgG rheumatoid factor could be demonstrated. These data demonstrate an unexpected cross reactivity of Fc fragment of IgG and VCA of EBV through the analysis of SF IC. PMID- 3025299 TI - Suppressor T cell growth and differentiation: evidence for induced receptors on suppressor T cells that bind a suppressor T cell differentiation factor. AB - T suppressor cell differentiation factor (TsDF) induces the differentiation of alloantigen-primed suppressor T cells (MLR-Ts) to expression of their effector function, i.e., to active TsF production. The initial activation stimulus to Ts is provided by alloantigen binding; after this binding, Ts are functionally responsive only for a period of hours to the additional stimulus provided by TsDF. The present studies addressed the possibility that MLR-Ts responsiveness to TsDF reflects the induced and transient display of TsDF-binding receptors. TsDF receptor expression was investigated by determining the capacity of TsDF responsive MLR-Ts to adsorb TsDF activity and to respond to that TsDF pulse by TsF production. Primed Ts populations that were alloantigen restimulated for 8 hr adsorbed TsDF in a cell dose-dependent fashion and produced TsF in response to that adsorption, whereas alloantigen-stimulated naive cells or primed but nonrestimulated cells neither responded to nor bound TsDF. Primed and restimulated L3T4-Ly-2+ but not L3T4+-Ly-2--enriched T cells bound TsDF. TsDF adsorption was saturable and time and temperature dependent. Glutaraldehyde fixation did not prevent TsDF adsorption by restimulated MLR-Ts, whereas pronase treatment abolished their TsDF-binding capacity. Kinetic analyses demonstrated that the capacity to bind TsDF developed rapidly after alloantigen reexposure, with maximal binding within 8 hr, followed by rapid decay with loss of TsDF binding by 36 hr. The kinetics of TsDF-induced TsF production correlated precisely with those of TsDF binding. These observations provide strong evidence that TsDF affects primed alloantigen-reactive Ts by interaction with antigen induced and transiently expressed cell surface receptors. TsDF-receptor binding is then the stimulus for expression of Ts effector function. PMID- 3025300 TI - Synthesis and release of leukotriene C4 by human eosinophils. AB - When human peripheral blood eosinophils isolated to 92.5% +/- 6.9 purity were stimulated with either the calcium ionophore A23187 or N-formyl-L-methionyl-L leucyl-L-phenylalanine (FMLP), immunoreactive leukotriene C4 (LTC4) was initially localized intracellularly and was subsequently released to the external medium in kinetically distinguishable steps. Eosinophils were stimulated with 2.5 microM A23187 in the presence of 20 mM L-serine, a hypochlorous acid scavenger that prevents the oxidative metabolism of sulfidopeptide leukotrienes. Total production of immunoreactive LTC4, the sum of intra- and extracellular LTC4, was complete within 5 to 10 min. At 5, 10, and 30 min, 65.9% +/- 15.2, 42.3% +/- 24.3, and 5.5% +/- 3.9, respectively, of the total amount of LTC4 measured remained intracellular as detected after the media and cells were separated and the latter was extracted with methanol. The time course for the intracellular synthesis and extracellular release of immunoreactive LTC4 from eosinophils pretreated with 5 micrograms/ml cytochalasin B and stimulated with 0.5 microM FMLP was like that obtained with ionophore, although the total LTC4 production was only approximately 10%. The identity of the intracellular LTC4 was confirmed by elution with reverse-phase high pressure liquid chromatography followed by scanning UV spectroscopy, radioimmunoassay, and bioassay. Eosinophils that were stimulated with A23187 in the absence of L-serine metabolized newly synthesized LTC4 to 6-trans-LTB4 diastereoisomers and subclass-specific diastereoisomeric sulfoxides that were identified only in the extracellular medium. Thus the response of purified eosinophils to two different stimuli demonstrates a transient intracellular accumulation of biologically active LTC4, the distinct extracellular release, and the apparent limitation of oxidative metabolism to the extracellular location. PMID- 3025301 TI - Modulation of the macrophage oxidative burst by Histoplasma capsulatum. AB - The production of reactive oxygen species by phagocytic cells is an important host defense against invading microorganisms. Because pathogens that achieve intracellular survival escape destruction by reactive oxidants, we investigated the relationship between the intracellular survival of H. capsulatum and the macrophage oxidative burst. H. capsulatum yeast failed to stimulate the release of reactive oxygen metabolites in unprimed murine macrophages despite extensive phagocytosis of the microorganisms. This effect was observed with live as well as heat-killed fungi over a wide range of yeast-to-macrophage ratios. Preincubation of murine macrophages with heat-killed H. capsulatum (but not with latex spheres), followed by incubation with unopsonized zymosan, resulted in inhibition of oxidative burst triggering without inhibition of zymosan phagocytosis. Ingestion of H. capsulatum yeast opsonized with the cognate mouse antibody resulted in significant oxidant release, suggesting that suppression of the respiratory burst may be circumvented through Fc-mediated phagocytosis. PMID- 3025302 TI - Human neonatal keratinocytes have very high levels of cellular vitamin A-binding proteins. AB - Since cellular retinol- and retinoic acid-binding proteins (CRBP and CRABP) mediate the effects of vitamin A on epidermal differentiation, the levels of these binding proteins were measured in the epidermal and dermal layers of newborn, human foreskin as well as in primary cultures of keratinocytes and fibroblasts from these layers. Ligand binding assays with saturating concentrations of all trans-[3H]retinol or of all trans-[11-3H]retinoic acid were used to quantitate amounts of binding proteins in cytosols prepared from these skin layers or cultured cells. The epidermal levels of CRABP and CRBP (60.9 +/- 14.4 and 7.3 +/- 1.7 pmol per mg cytosol protein, respectively) were markedly higher than that reported for adult epidermis but were comparable to levels in keratinocytes cultured from neonatal foreskin epidermis (61.8 +/- 7.8 and 10.7 +/ 2.5, respectively). The levels of CRABP were much lower in the foreskin dermis than in the epidermis and the levels measured in the fibroblasts cultured from this dermis were consistent with the dermal levels. However, CRBP levels in cultured dermal fibroblasts were very low and could not account for the dermal CRBP levels, suggesting that another dermal cell type has high levels of CRBP. PMID- 3025303 TI - Absence of antibodies to human immunodeficiency virus in homosexual, hemophiliac, and heterosexual men in Budapest, Hungary in 1983-1984. PMID- 3025304 TI - Human enteric coronaviruses: further characterization and immunoblotting of viral proteins. PMID- 3025305 TI - Nucleic acid sequences of cytomegalovirus in cells cultured from human arterial tissue. PMID- 3025306 TI - Humoral and cell-mediated immunity in neonates with herpes simplex virus infection. AB - Fifty-nine neonates with herpes simplex virus (HSV) infection were evaluated with use of assays for neutralizing antibody (NAb), lymphocyte transformation (LT), alpha interferon production, and virus-specific antibody (immunoblots). Infants with disseminated disease or onset in the first week of life were more likely to lack NAb. Patients treated with vidarabine were more likely than those treated with acyclovir to develop a fourfold rise in NAb titer. Infants with encephalitis showed a broader spectrum of IgG and IgM antibody reactivity against HSV proteins by immunoblotting than did those who had earlier onset of mucocutaneous illness. Only 10 of 33 infants had HSV-specific LT, compared with eight of eight adults with primary HSV. Neonates with positive LT were more likely to show a fourfold rise in NAb titer. In vitro alpha interferon production was diminished in infants, compared with values in adults. PMID- 3025307 TI - Detection of antibodies to herpes simplex virus in the cerebrospinal fluid of patients with herpes simplex encephalitis. AB - Cerebrospinal fluid (CSF) and serum specimens from patients with presumed herpes simplex encephalitis (HSE) were characterized for antibodies to herpes simplex virus (HSV) by immunoblot and other immunoassays. Specimens from patients proven to have HSE (biopsy-proven) were compared with those from patients with other diseases diagnosed by brain biopsy (biopsy-negative for HSV). Immunoblot of CSF demonstrated that antibodies to HSV-specific polypeptides, particularly to glycoprotein B, were present in a matched serum specimen. When purified glycoprotein B was used to detect CSF antibodies, 34 of 35 specimens from biopsy proven patients were reactive (97% sensitivity) compared with only six of 22 specimens obtained from biopsy-negative patients (73% specificity). If leakage of antibodies to a marker virus (adenovirus) was determined and controlled, the specificity increased to 100%. These diagnostic assays provide useful tools for the retrospective assessment of CSF specimens obtained from patients with presumed HSE. PMID- 3025308 TI - Antiviral activities of human monoclonal antibodies to herpes simplex virus. AB - Hybridomas producing human monoclonal antibodies (MAbs) against herpes simplex virus (HSV) were established by fusing human tonsillar lymphocytes with mouse myeloma cells. Three hybridomas have been stably producing MAbs for more than 16 months. All three MAbs--H1, H2, and H3--were of the IgG1 isotype and recognized the gB glycoprotein of HSV types 1 and 2 (HSV-1 and HSV-2). MAbs H2 and H3 not only bound to the surface membrane of HSV-infected cells but also neutralized both HSV-1 and HSV-2, whereas MAb H1 had neither activity. In mouse infection experiments, MAbs H2 and H3 showed a potent protective effect against HSV-1 infection, whereas MAB H1 was less protective. Furthermore, the development of zosteriform skin lesions in athymic nude mice was suppressed by administering MAb H2. These results suggest that human MAbs might provide passive immunization against HSV infections in humans. PMID- 3025309 TI - AIDS and antibodies to human immunodeficiency virus (HIV) in children and their families. AB - Infection with human immunodeficiency virus (HIV, previously known as HTLV III/LAV) documented by a sensitive, specific immunoblotting (western blot) technique is described in 14 children with symptoms of AIDS or AIDS-related complex. For serodiagnosis of HIV infection, immunoblots blocked with milk were more sensitive than enzyme-linked immunosorbent assays or immunoblots blocked with gelatin. One or both parents of 13 of these children abused intravenous drugs. Sixteen of 17 parents of the affected children but only one of eight siblings living in the same household were positive for antibody to HIV. All siblings had experienced infection and acquired antibodies to Epstein-Barr virus, which is thought to spread by saliva. In contrast to their HIV-infected parents, children with AIDS or AIDS-related complex were less likely to have decreased numbers of circulating T4 cells, and their sera recognized fewer HIV polypeptides on western blots. PMID- 3025310 TI - [Pathogenesis of atrial myocarditis in monkeys inoculated with Cb1oxsackie virus B3]. PMID- 3025311 TI - [Acute respiratory infections among infants and children in Japan. The first report: Virus isolation employing a new microplate method]. PMID- 3025312 TI - [A possible physiological role of NDP-kinase in cell differentiation and cell proliferation]. PMID- 3025313 TI - [An ultrastructural study of the localization of alkaline phosphatase and Na-K ATPase in the labyrinth from the IUGR rat]. AB - To elucidate the pathogenesis of intrauterine growth retardation (IUGR), the ischemic placentae produced by uterine artery and vein ligation were morphologically and histochemically investigated in rats. In the ischemic condition, fetal growth was remarkably retarded. The labyrinthi were atrophic and trophoblasts were degenerated ultrastructurally in association with mitochondrial swelling, increased microvesicles, dense bodies and fibrin deposition in the dilated intercellular spaces. Alkaline phosphatase and Na-K ATPase were localized on the plasma membrane in control rats. Na-K ATPase activity was observed in the basal side of cytotrophoblasts. However, the enzyme activities were decreased in the ischemic condition. The reaction products of horseradish peroxidase were present in the dilated intercellular spaces and increased cytoplasmic vesicles. These results suggest that in the ischemic condition, IUGR might be induced by the plasma membrane dysfunction following decreased enzyme activities and irregular distribution of horseradish peroxidase. PMID- 3025314 TI - [Immunohistological study on malignant and premalignant lesions of the uterine cervix associated with herpes simplex virus]. AB - Herpes Simplex Virus 2 (HSV-2) has become the object of public attention as an etiological cause of cervical cancer. Uneven distribution of HSV antigen in tissue was examined dy dyeing tissue materials of cervical intraepithelial neoplasia by the method called peroxidase-antiperoxidase using HSV-2 antibody. The positive rate in the control group was 10.3%, while it was 10.7% in the mild dysplasia group, 17.5% in the moderate dysplasia group, 25.5% in the severe dysplasia group, 31.3% in the carcinoma in situ (CIS) group and 41.4% in the cervical cancer group. The positive rate in the cervical cancer group showed a significant difference from the control, mild displasia, moderate dysplasia and severe dysplasia groups (p less than 0.01). The rate in the CIS group demonstrated a significant difference from the control and mild dysplasia groups (p less than 0.05, p less than 0.01). The rate in the severe dysplasia group showed a significant difference from the control and mild dysplasia groups (p less than 0.05, p less than 0.01). The positive rate in the cervical cancer group in patients 40 approximately 49 years of age was significantly higher than that in the control and mild dysplasia groups (p less than 0.01). It also showed a significant difference from the moderate dysplasia and severe dysplasia groups (p less than 0.05). The rate in the cervical cancer group in patients aged 50 approximately 59 years showed a significant difference from the control, moderate dysplasia and mild dysplasia groups (p less than 0.05, p less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3025316 TI - [Experimental conditions for fluorapatite formation on tooth enamel using the silica gel method "in vivo". A study of fluorapatite crystallized by a modified silica gel method]. PMID- 3025315 TI - [Immunobiological activity of 41K protein (porin) derived from the cell envelope of Fusobacterium nucleatum]. PMID- 3025317 TI - [Alcoholic nervous disorders]. PMID- 3025318 TI - Calcium-induced degeneration of the cytoskeleton in monkey and human peripheral nerves. AB - Biopsy specimens of human and monkey peripheral nerves, when incubated in calcium containing media, showed a loss of neurofilaments and microtubules with replacement by granular debris. Cytoskeletal structures remained intact when incubated in calcium-free media. Disruption of neurofilaments and microtubules in calcium containing media was inhibited by the thiol protease inhibitor, leupeptin. Similar incubations of excised Pacinian corpuscles revealed evidence of early terminal axon degeneration in the presence of calcium. These data substantiate the hypothesis that neural cytoskeletal degradation in primates and in man is calcium-mediated. PMID- 3025319 TI - Degenerated muscle grafts used for peripheral nerve repair in primates. AB - The basement membrane matrix of skeletal muscle has a tubular configuration resembling that of peripheral nerves. Grafts made of autogenous skeletal muscle denatured by freezing and thawing were used to repair the ulnar nerve in marmosets. By six months, normal hand function had returned and the grafts were shown to transmit normal compound extracellular action potentials in both directions. Morphological examination of the grafts and distal nerves revealed normal axon numbers and axon maturity. Myelination in the graft was found to take place more slowly than in the distal nerve segment. It is suggested that such grafts might be of use in the repair of human peripheral nerves. PMID- 3025321 TI - Synovial sarcoma of the hand--a literature study. AB - Ninety cases of synovial sarcoma of the hand, including eight case reports have been described during the last fifty years, (1934-1984). 8.5% of all synovial sarcomata involve the hand, and affect predominantly individuals under the age of thirty. The five-year survival of these cases was 18% and the ten-year survival 9%. PMID- 3025320 TI - Congenital anomalies of the upper limb among the Japanese in Sapporo. AB - Nine hundred and forty three patients with nine hundred and fifty-five congenital upper limb anomalies are classified according to the Swanson's classification. Relative incidence of each type of anomaly, sex incidence, affected side, associated abnormalities and familial history are analysed. The occurrence of congenital upper limb anomalies is also compared with Chinese and Western studies. The characteristic feature in this series is that the incidence of typical syndactyly is not as high as expected. Two problems of Swanson's classification encountered in this analysis are discussed. One of the problems is the relation between polydactyly, syndactyly and typical cleft hand. The other is the relation between brachysyndactyly, atypical cleft hand and transverse deficiency. PMID- 3025322 TI - Calcifying synovial sarcoma. AB - The presence of a calcified mass in the soft tissue may be the result of any number of processes. Post-traumatic changes and metabolic alterations may both lead to such calcification. The presence of calcification is much less commonly seen in malignant neoplasms. A case of calcifying synovial sarcoma with unusual features was recently seen. PMID- 3025323 TI - Evidence that chylomicron remnants and beta-VLDL are transported by the same receptor pathway in J774 murine macrophage-derived cells. AB - To characterize lipoprotein uptake by macrophages, we studied J774 murine macrophage-derived cells. Uptake of 125I-labeled beta-VLDL and 125I-labeled chylomicron remnants was saturable, specific, and of high affinity. Maximal specific uptake and the concentration at which half-maximal uptake occurred were similar for both beta-VLDL and chylomicron remnants. Specific uptake of 125I labeled chylomicrons was only 1/5 that of the other two lipoproteins. Cholesterol loading decreased 125I-labeled chylomicron remnant and 125I-labeled beta-VLDL uptake by 25%. Chylomicron remnants and beta-VLDL were equipotent in cross competition studies; acetyl-LDL did not compete, and human LDL was a poor competitor. Although the amounts of cell-associated lipoproteins were similar, beta-VLDL and chylomicron remnants had different effects on cellular lipid metabolism. beta-VLDL produced a threefold stimulation while chylomicron remnants caused a decrease in [3H]oleate incorporation into cholesteryl ester. beta-VLDL had no effect while chylomicron remnants caused a threefold increase in [3H]oleate incorporation into triacylglycerol. beta-VLDL produced a 44% suppression and chylomicron remnants produced a 78% increase in HMG-CoA reductase activity. In summary, J774 macrophages express a receptor site that recognizes both beta-VLDL and chylomicron remnants; however, these lipoproteins exhibit strikingly different effects on intracellular lipid metabolism. PMID- 3025324 TI - Effects of dietary retinoic acid on cellular retinol- and retinoic acid-binding protein levels in various rat tissues. AB - A study was conducted to explore the effects of retinoic acid, fed to retinol deficient rats, on the tissue distribution and levels of cellular retinol-binding protein (CRBP) and cellular retinoic acid-binding protein (CRABP). Sensitive and specific radioimmunoassays were employed to measure the levels of both CRBP and CRABP. Two groups of six male rats each were fed a purified retinoid-deficient diet supplemented with either: i) retinyl acetate (control group); or ii) retinoic acid (30 mg/kg diet) (retinol deficient-retinoic acid group). The retinoic acid supplementation was begun after 38 days on the retinoid-deficient diet alone, and was continued for 52-54 days. Analysis of the data indicated that only the CRBP level of the proximal epididymis in the retinol-deficient/retinoic acid group differed significantly from (was lower than) the corresponding control level, at the 1% confidence level. CRABP tissue levels did not differ significantly between the two groups. Thus, a moderately large intake of retinoic acid, as the only source of retinoids, had very little effect on the tissue distribution or levels of either its own cellular binding protein (CRABP) or of CRBP. This study provides further information showing that the tissue levels of the cellular retinoid-binding proteins are highly regulated and maintained in rats, even in the presence of marked changes in retinoid nutritional status. PMID- 3025325 TI - Juvenile nasopharyngeal angiofibroma presenting as a facial swelling. A case report. AB - Juvenile nasopharyngeal angiofibroma is a rare benign neoplasm occurring almost exclusively in adolescent males. When it is confined to the nasopharynx, surgery is often curative. In 20% of cases, there is intracranial extension, and radiotherapy may be used to avoid the risk of life-threatening haemorrhage. The authors report an unusual case which presented with a swelling of the cheek and an abducens nerve palsy. The lesion extended from the nasopharynx across the pterygomaxillary fissure, as well as intracranially. Radiotherapy was given, and the patient remains disease-free after one year. PMID- 3025326 TI - The characteristics of beta-adrenergic binding sites on pancreatic islets of Langerhans. AB - The sympathetic nervous system is believed to play a part in the control of insulin release from the pancreatic islets of Langerhans. Stimulation of alpha adrenoceptors is thought to inhibit the release of insulin whereas stimulation of beta-adrenoceptors enhances insulin release. The present experiments were conducted to establish the existence of beta-adrenergic receptors on guinea-pig and rat islet cells and to quantify them using the selective beta-adrenergic ligands [3H]dihydroalprenolol (DHA) and [125I]cyanoiodopindolol (CYP). Guinea-pig islets had 62 fmol beta-adrenoceptors/mg protein using [3H]DHA, corresponding to 43,700 binding sites/cell and 25 fmol beta-adrenoceptors/mg protein using [125I]CYP, corresponding to 17,400 sites/cell. Rat islet cells were found to have 4.6 fmol beta-adrenoceptors/mg protein using [125I]CYP, corresponding to 7200 sites/cell. Adenylate cyclase activation exhibited a positive dose-response relationship when exposed to the beta-adrenoceptor agonist isoprenaline, with a maximum response (190 +/- 21% above basal) at 10 mumol isoprenaline/l. This response was abolished with 1 mumol/l of the beta-adrenergic antagonist l alprenolol. Insulin secretion in the presence of 10 mmol glucose/l, but in the absence of the alpha-adrenoceptor blocker phentolamine, was not affected by 10 mumol isoprenaline/l. However, perifusion experiments showed that secretion of insulin from isolated rat islets in the presence of 10 mmol glucose/l was significantly increased (332%) by 10 mumol isoprenaline/l in the presence of 10 mumol phentolamine/l. These results suggest that binding of selective radiolabelled ligands occurs to beta-adrenergic receptors on the B cell surface of the islets of Langerhans, and that these receptors are functionally coupled to insulin secretion through modulation of adenylate cyclase activity. PMID- 3025327 TI - The distribution of immunoreactive alpha-melanocyte-stimulating hormone cells in the adult human pituitary gland. AB - It has been suggested that a proportion of the adenomas and the nodular hyperplasia of cells in the pituitary gland in cases of Cushing's disease are derived from cells of the pars intermedia rather than the pars anterior. The evidence can be summarized as follows: the posterior site of adenoma or nodular hyperplasia in the pituitary, the innervation of cells and the suppressive response to the dopamine agonist bromocriptine in vivo or to dopamine in vitro. All these observations infer analogy with cells of the pars intermedia of other species, which are controlled by direct neural tonic dopaminergic inhibition. The adult human pituitary gland, however, does not possess a morphologically distinct pars intermedia, due to regression of the rudimentary fetal pars intermedia after birth, with mixing of cells into the pars anterior and pars nervosa. Since cells of the pars intermedia characteristically synthesize alpha-MSH, we have studied this peptide in order to assess the occurrence and distribution of intermedia derived cells in the adult human pituitary. Sections from 100 pituitaries, removed at autopsy, were stained by an indirect immunoperoxidase technique using non-cross-reacting antisera specific for alpha-MSH and ACTH. Immunoreactive alpha MSH (IR-alpha-MSH) cells were found in a total of 97 specimens. Of these, only ten cases showed a marked concentration of IR-alpha-MSH cells in the zona intermedia. In the majority of pituitaries, IR-alpha-MSH cells were more commonly seen in the pars anterior than in the zona intermedia; in 41 cases, IR-alpha-MSH cells were completely absent from the zona intermedia.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3025328 TI - The pituitary adrenocorticotropes originate from neural ridge tissue in Xenopus laevis. AB - A series of grafting experiments was conducted to determine pituitary origins prior to brain tube closure in Xenopus laevis. Extirpation experiments indicated that the ventral neural ridge (VNR) tissue of stage-18+ embryos was essential for pituitary development. Bolton-Hunter reagent was used to label stage-18+ VNR tissue with 125I, and this tissue was then returned to the donor and its subsequent ontogenesis followed. Labelled tissue was ultimately found in the ventral hypothalamus, the ventral retina, and the anterior pituitary. Using immunocytochemical techniques with antisera to adrenocorticotropin (ACTH), it was found that some of the VNR-derived cells were corticotropes. A region of the nucleus infundibularis which was radioactive labelled also gave ACTH-positive immunoreaction. This might indicate that some ACTH-containing neurones of the hypothalamus are VNR in origin. We suggest that stage-18+ VNR is the site of attachment of brain and anterior pituitary ectoderm. Part of this adherence point is eventually incorporated into the anterior pituitary and will form corticotropes. It is concluded that the ventral retina, the preoptic region of the hypothalamus, some hypothalamic ACTH-immunoreactive cells, and the most anterior portion of the adenohypophysis are all ventral neural ridge in origin. PMID- 3025329 TI - Retinoic acid-binding protein in the chick limb bud: identification at developmental stages and binding affinities of various retinoids. AB - The application of retinoic acid (RA) to the developing chick limb bud causes 6 digit double posterior limbs to form instead of the normal 3-digit limb. As an attempt to begin a molecular analysis of this phenomenon we have identified and characterized a soluble cytoplasmic receptor for RA, namely cytoplasmic retinoic acid-binding protein (CRABP), from the cells of the chick limb bud. It is present from stages 20-35 at similar levels and has an apparent Kd of 140-280 nM. In competition experiments with other retinoids Ro 13-7410 was found to be the most effective at competing for sites on CRABP followed by all-trans-RA, 13-cis-RA, Ro 10-1670 and retinal. Retinol, retinyl palmitate, retinyl acetate, etretinate and arotinoid showed low or no affinity for CRABP. Specificity for binding was thus demonstrated since analogues with an acid end group competed effectively, the aldehyde competed less effectively and the ester or alcohol groups did not compete. At the concentration of RA that needs to be administered to cause duplications in the pattern of the limb bud, we estimate that 4% of the CRABP present in the limb bud has RA bound. The similarities between steroid receptors in the mediation of steroid hormone action and CRABP in the mediation of RA action is discussed. In this regard we note that while there are 10(4) steroid receptors per cell in other cell types we estimate that there are about 10(5) RA receptors per cell in the chick limb bud. PMID- 3025330 TI - Leishmania promastigotes are recognized by the macrophage receptor for advanced glycosylation endproducts. AB - In this paper we demonstrate the involvement of the macrophage receptor for advanced glycosylation endproducts (AGE) in the phagocytosis of Leishmania major promastigotes. Blocking of this receptor with the ligand, AGE-BSA, leads to a 50% decrease in phagocytosis relative to controls, and a comparable decrease in the respiratory burst. The inhibition of phagocytosis by AGE-BSA was specific to leishmania. The binding of zymosan or C3bi-RBC and the phagocytosis of IgG-RBC or latex beads was not affected by the presence of AGE-BSA. Blocking of both the AGE receptor and CR3 decreases leishmania binding by nearly 90%, and reduces the respiratory burst by 80%, indicating that the two receptors account for the bulk of L. tropica promastigote recognition and uptake by the macrophage. PMID- 3025332 TI - Autoantibodies to neurofibrillary tangles and brain tissue in Alzheimer's disease. Establishment of Epstein-Barr virus-transformed antibody-producing cell lines. AB - Multiple EBV-transformed B cell lines were established from five patients with a clinical diagnosis of Alzheimer's disease (AD) and six age-matched controls. The supernatants were screened for antibody activity against SDS-treated isolated neurofibrillary tangles (NFT). Reactive supernatants were identified from both the AD and control group. The frequencies of anti-NFT antibody-secreting lines were 6.3 and 1.6% for the AD and the control groups, respectively. A proportion of these supernatants also stained NFT in situ and neurons and/or glia in sections of the frontal and the temporal cortexes of autopsied AD and normal brains, as well as cells from three cell lines (HeLa, fibroblast, and neuroblastoma). Several patterns of staining were revealed by these supernatants, indicating different reactive antigens. One supernatant stained NFT and astrocytes in sections from AD brains. It did not stain sections from two normal brains. This cell line is the result of the immortalization of a circulating B cell making antibody specific for an antigen in AD. The present approach may provide new insights in the pathogenesis of AD. PMID- 3025334 TI - Cyclic AMP stimulation of Cl- secretion by the opercular epithelium: the basolateral chloride uptake mechanism. AB - The isolated, short-circuited opercular epithelium of Fundulus heteroclitus, secretes Cl- by a mechanism dependent on the presence of serosal Na+ and inhibited by bumetanide and furosemide. Under serosal Na+-free conditions the active Cl- secretion is abolished. However, subsequent elevations of intracellular cyclic AMP (cAMP) levels with isoproterenol or forskolin stimulated Cl- secretion markedly. This stimulation was unaffected by SITS, DIDS, methazolamide, and HCO-3-free solutions, but was blocked by furosemide and bumetanide. Determinations of relative intracellular 36Cl- levels showed a Na+ dependence of intracellular 36Cl- in epithelia not stimulated by isoproterenol and a Na+ independence of intracellular 36Cl- in isoproterenol stimulated epithelia. In both conditions, the intracellular 36Cl- was bumetanide sensitive. The results indicate that cAMP stimulation of Cl- secretion can occur by a Na+ independent, loop diuretic-inhibitable mechanism, which may be operative even in the presence of Na+. Whether this is a separate Cl- uptake mechanism or a cAMP induced alteration in the normal Na+-dependent mechanism could not be determined. In either instance, an alternative to the Na+ gradient as a source of energy for Cl- uptake into the cell across the basolateral membrane is required. PMID- 3025333 TI - Reversibility of gelsolin/actin interaction in macrophages. Evidence of Ca2+ dependent and Ca2+-independent pathways. AB - We have developed an immunoadsorption technique for quantitating EGTA-resistant gelsolin/actin complexes in macrophages extracted with Triton X-100. We report here that the proportion of gelsolin complexed irreversibly to actin is low in freshly harvested macrophages. The amount of the EGTA-resistant complex increases spontaneously during incubation of the cells in suspension at 37 degrees C, or after exposure to the Ca2+ ionophore ionomycin. On the other hand, exposure of suspended cells to the chemotactic oligopeptide, FMLP, or plating of the cells onto tissue culture dishes causes the EGTA-resistant complex to dissociate rapidly. Plating even prevents Ca2+ ionomycin-treated cells with elevated intracellular Ca2+ from inducing this complex. Therefore, our results suggest that macrophages possess a mechanism, not directly involving Ca2+, for dissociating actin/gelsolin EGTA-resistant complexes. This mechanism may be a Ca2+-independent signal for leukocyte activation. PMID- 3025331 TI - Role of the adherence-promoting receptors, CR3, LFA-1, and p150,95, in binding of Histoplasma capsulatum by human macrophages. AB - The principal host cell of H. capsulatum (Hc) is the M phi within which the pathogenic yeast phase of the fungus multiplies during active disease. The initial interaction between Hc yeasts and M phi therefore is a crucial step in the pathogenesis of histoplasmosis. In the present study, we have identified the major receptor mechanism that mediates the attachment of unopsonized Hc yeasts to human monocyte-derived M phi from peripheral blood. Binding of Hc yeasts by M phi is rapid, temperature dependent, and requires both Ca and Mg ions for optimum activity. Recognition of Hc yeasts does not require Fc receptors, mannosyl/fucosyl receptors, beta-glucan receptors, or secretion of C3 by M phi. Studies were performed on the effect of down regulating specific receptors of the CR3/LFA-1/p150,95 adherence-promoting protein family from the apical portion of M phi to determine the effects upon binding of Hc yeasts. Anti-beta chain mAbs that recognize all three of these proteins blocked binding of yeasts. However, removal of individual receptors with antibodies against the alpha polypeptides caused negligible depression of binding, and removal of any pair caused only modest depression. Thus, each of the members of the CR3/LFA-1/p150,95 family is independently capable of binding Hc. The delineation of this new mechanism for nonopsonic recognition by M phi that is exploited by Hc yeasts will aid in future studies to identify the Hc ligand, to elucidate the stoichiometry of CR3/LFA 1/p150,95 binding, and to determine triggering mechanisms for release of toxic oxygen metabolites. PMID- 3025335 TI - Properties of in vitro recombinant derivatives of pJV1, a multi-copy plasmid from Streptomyces phaeochromogenes. AB - The 10.8 kb plasmid pJV1, isolated from Streptomyces phaeochromogenes, has a high copy number (about 150) and a broad host range among Streptomyces spp. Several pJV1 derivatives carrying the thiostrepton resistance gene (tsr) of S. azureus were made. One derivative, pWOR191, was shown to promote its own transfer and to mobilize chromosomal markers in S. lividans. Another derivative, pWOR109, was non transmissible. Deletion in vitro of a segment of pWOR109 gave pWOR120 (5.6 kb), which has single BamHI and Bg/II sites shown to be capable of accepting 'foreign' DNA such as a previously cloned S. antibioticus DNA fragment encoding tyrosinase, giving vectors (pWOR125, pWOR126) with properties resembling the well-established multicopy vector pIJ702. Shuttle vectors capable of functioning in both S. lividans and Escherichia coli were also constructed. The region of pJV1 essential for replication and maintenance was localized to a 2.5 kb segment. Stable maintenance of pWOR109 and pWOR120 was observed in the presence of derivatives of pIJ101, the progenitor of pIJ702. PMID- 3025336 TI - Effect of mitochondrial cytochromes and haem content on cytochrome P450 in Saccharomyces cerevisiae. AB - It is well established that the mitochondrial and the microsomal cytochromes in Saccharomyces cerevisiae are regulated differently. Mutations affecting the mitochondrial cytochromes aa3 or c had no effect on the concentration of the microsomal cytochrome P450 even during haem limitation. Moreover, a defect in the cytochrome P450 gene did not affect mitochondrial cytochromes. However, a regulatory mutation present in strain SG1 decreased both mitochondrial and microsomal cytochrome contents. This mutation also affected the intracellular haem concentration. The haem precursor 5-aminolaevulinate increased both mitochondrial and microsomal cytochrome contents. Our results indicate that carbon source and haem concentration are involved in the regulation of cytochrome P450. PMID- 3025337 TI - Separation of Mycobacterium gadium from other rapidly growing mycobacteria on the basis of DNA homology and restriction endonuclease analysis. AB - DNA was isolated from Mycobacterium gadium with high purity. Its G + C content was between 64 and 67 mol%. The homology of M. gadium DNA with DNA from three other rapidly growing mycobacteria was less than 22%, which indicates that M. gadium is a discrete genomic species. Analysis of the DNAs with restriction endonucleases supported this finding. PMID- 3025338 TI - A multi-resistance plasmid isolated from commensal Neisseria species is closely related to the enterobacterial plasmid RSF1010. AB - pFM739, an R plasmid from Neisseria sicca that encodes penicillin, streptomycin and sulphonamide resistance, and the enterobacterial IncQ(P-4) plasmid RSF1010, which encodes streptomycin and sulphonamide resistance, were incompatible, and were mobilized by the same conjugative plasmids. Restriction mapping confirmed a high degree of similarity between both R plasmids; pFM739 carried DNA fragments corresponding to the known replication and resistance regions of RSF1010. pFM739 also carried an extra segment with the same restriction map as that described for the beta-lactamase-coding region of transposon Tn3. It is suggested that the R plasmids isolated from commensal Neisseria sp. could have resulted from transposition of a Tn3-like genetic element to an RSF1010-like plasmid, and that they contain deletion derivatives of transposon Tn3. PMID- 3025339 TI - Isolation and characterization of a herpes simplex virus type 1 mutant containing a deletion within the gene encoding the immediate early polypeptide Vmw110. AB - Transfection experiments with plasmids containing immediate early (IE) genes of herpes simplex virus type 1 (HSV-1) have previously demonstrated a role for the IE polypeptide Vmw110 (ICP0) in stimulating expression from plasmid-encoded early gene promoters. To gain further insights into the function of Vmw110 we isolated a deletion mutant specifying a truncated form of the polypeptide which had been shown to be inactive in transfection assays. This mutant, dl1403, contained a 2 kb deletion within both the TRL and IRL copies of the Vmw110 gene, and encoded a polypeptide consisting of the original N-terminal 105 amino acids followed by 56 amino acids specified by a reading frame not used by Vmw110. dl1403 was able to replicate and produce plaques on baby hamster kidney (BHK) cells but the yield of infectious virus was 20- to 100-fold lower than obtained with wild-type HSV-1. Surprisingly, comparison of polypeptide synthesis, DNA replication and DNA encapsidation in cells infected with 5 p.f.u./cell dl1403 or wild-type HSV-1 revealed no significant differences. In addition similar numbers of particles were produced in cells infected with the two viruses, resulting in stocks of dl1403 exhibiting significantly higher particle/p.f.u. ratios. The efficiency of plaquing of dl1403 was greatly reduced in Vero and human foetal lung cells compared with BHK cells, but following infection with 5 p.f.u./cell similar yields of infectious virus were obtained from all three cell lines. Marker rescue experiments verified that the reduced yield of dl1403 in BHK cells was a consequence of the deletion within the Vmw110 gene. The results suggest that the effect of this deletion is manifest primarily at low multiplicities of infection and can be largely overcome by increasing the virus dose. PMID- 3025340 TI - Occurrence of novel small RNAs with concomitant inhibition of host cellular U small nuclear RNA synthesis in Vero cells infected with herpes simplex virus type 1. AB - In eukaryotic cells, small nuclear and small cytoplasmic RNAs (sn- or scRNAs) are associated with distinct proteins, forming ribonucleoproteins (snRNPs or scRNPs). In the present study we analysed the protein composition as well as the small RNA pattern in non-infected and herpes simplex virus type 1 (HSV)-infected Vero cells. We found that concomitantly with the shut-off of host cell mRNA synthesis, synthesis of U-snRNAs was stopped. Due to their stability, however, U-snRNAs were still present in cells 36 h after HSV infection. Besides these RNAs, two novel small RNAs which we termed HVR1 and HVR2 were detected in infected cells. On the basis of their relative mobilities in urea gels, the apparent chain lengths of these newly synthesized RNAs were determined to be 255 and 154 nucleotides respectively. The small RNA-binding proteins Sm, RNP, Ro and La were found to increase up to 15-fold after HSV infection. The data presented suggest that new, virus-coded small RNPs are synthesized which might play a role in the maturation and regulation of HSV-coded RNA transcripts. PMID- 3025341 TI - Human cytomegalovirus replicates in primary human bone marrow cells. AB - As an attempt to elucidate further the pathogenesis of human cytomegalovirus (HCMV) infection the replication of HCMV in primary human bone marrow cells (BMC) has been investigated. It was found that BMC held in culture in general were susceptible to HCMV infection. Compared to human embryonic lung cells, however, the replicative cycle of HCMV AD169 in BMC as determined by the analysis of viral protein and DNA synthesis was delayed and productive virus infection was restricted to a subset of BMC not exceeding 21% of the total cell population. Both of these phenomena may explain the short-term persistence of HCMV in BMC cultures which was observed over 3 months. By experiments with specifically enriched and depleted cell populations and by indirect double immunofluorescence experiments we found that both bone marrow fibroblasts and a subset of bone marrow stem cells supported productive virus infection. The finding that HCMV replicates in early stem cells of the human bone marrow may explain important aspects of the pathogenesis of HCMV infection including the presence of HCMV in peripheral blood leukocytes. PMID- 3025342 TI - Human cytomegalovirus persistent infection in a human central nervous system cell line: production of a variant virus with different growth characteristics. AB - The susceptibility of human central nervous system cell lines to human cytomegalovirus (HCMV) and the fate of infected cultures were studied. Significant amounts of infectious progeny virus were produced in 118MGC glioma and IMR-32 neuroblastoma, but not in KGC oligodendroglioma cells when the cultures were infected with wild-type virus (HCMVwt) at an m.o.i. of 10 p.f.u. per cell. Further passage of infected 118MGC cells resulted in the establishment of a long-term persistent infection. This infection, designated 118MGC/Towne, continuously produced infectious virus (HCMVpi) with titres ranging from 10(2) to 10(5) p.f.u./10(6) cells up to 360 days post-infection (corresponding to 50 subcultures). Since no temperature-sensitive mutants, defective interfering particles or interferon-like activity were found in the 118MGC/Towne cultures, maintenance of the persistent infection seemed to be due to a balance between the release of infectious virus and the growth of uninfected cells. The HCMVpi produced in long-term persistently infected cultures was shown to be different from the HCMVwt originally used to infect by the following characteristics: HCMVpi replicated slowly and yielded lower amounts of progeny virus than HCMVwt; HCMVpi induced a 73,000 mol. wt. immediate early protein that was not synthesized in HCMVwt-infected cells; HCMVpi had a different DNA structure from that of HCMVwt. These results suggest that HCMVpi is a slower growing variant of HCMVwt and probably plays an important role in the maintenance of the persistent infection. PMID- 3025343 TI - Kinetics, tissue specificity and pathological changes in murine rotavirus infection of mice. AB - Mice that did not contain antibodies to rotavirus were orally infected with murine rotavirus (EDIM strain) and observed over 7 days. As judged by ELISA, only the small intestine was infected, not the colon. The infection was biphasic, viral antigen peaks being observed at 48 h and approximately 120 h post infection. Clinically evident diarrhoea was maximal at 72 h. Virus in the upper, middle and lower regions of the small intestine was mainly tissue-associated; most virus was found in the middle small intestine. Two peaks (48 h and 120 h post-infection) of virus antigen were observed in the colon, but these corresponded to luminal, not tissue-associated viral antigen. Only enterocytes in the upper two-thirds of villus epithelia were infected as judged by fluorescent antibody analysis and transmission electron microscopy. Scanning electron microscopy revealed morphological appearances not hitherto correlated with the progress of the infection: villus tips were convoluted, corresponding to the shedding of virus-infected cells but the lower regions of infected villi were shrunken and considerably narrowed compared to tips. PMID- 3025344 TI - Characterization of the mutations responsible for the electrophoretic mobility differences in the NS proteins of vesicular stomatitis virus New Jersey complementation group E mutants. AB - Temperature-sensitive (ts) mutants of vesicular stomatitis virus, New Jersey serotype, classified in complementation group E contain lesions in the NS gene, which manifest as marked electrophoretic mobility differences of the mutant NS proteins in SDS-polyacrylamide gels. We have cloned full-length cDNA copies of the mutant NS mRNAs, and have determined their nucleotide sequences. tsE1 and tsE3 had single nucleotide changes, and tsE2 had two nucleotide changes, compared to the wild-type NS gene. Three of the mutations were clustered in a region of 18 nucleotides. All the nucleotide differences resulted in amino acid substitutions, which in each case changed the charge of the amino acid concerned. Analysis of the wild-type and mutant NS protein sequences by the method of Chou & Fasman indicated that single amino acid substitutions can radically alter the predicted secondary structure, and these data are discussed in relation to the observed electrophoretic mobility differences. PMID- 3025345 TI - Nucleotide sequence of the gene encoding the fusion glycoprotein of Newcastle disease virus. AB - The nucleotide sequence of the gene encoding the fusion (F) glycoprotein of the Beaudette C strain of Newcastle disease virus (NDV) has been determined from cDNA clones obtained from virion RNA. The gene is 1792 nucleotides long, including mRNA start and polyadenylation signals typical of paramyxoviruses. The single open reading frame encodes a polypeptide of 553 amino acids, with a predicted molecular weight of 59042. The F polypeptide has three regions of high hydrophobicity: an N-terminal signal peptide, the N terminus of F1 (known from protein sequencing) and a C-terminal membrane-spanning region by which the F glycoprotein is anchored to the membrane. The cleavage site of F0 is located in a highly basic region of the F polypeptide. Five potential asparagine-linked glycosylation sites are present in the amino acid sequence, of which one is in F2 and the others in F1. Comparison of the NDV F amino acid sequence to those from other paramyxoviruses reveals homology to Sendai virus, simian virus 5 and human respiratory syncytial virus. There is also limited homology between the N terminus of F1 of NDV and the N termini of HA2 of influenza viruses. Post translational modifications of the NDV F polypeptide are discussed in the light of information provided by the amino acid sequence. PMID- 3025346 TI - Sequence analysis of the P and C protein genes of human parainfluenza virus type 3: patterns of amino acid sequence homology among paramyxovirus proteins. AB - The complete nucleotide sequence of the P + C mRNA of human parainfluenza virus type 3 (PF3) was determined by sequencing cDNA, viral genomic RNA and mRNA. The P + C mRNA is 2009 nucleotides in length, exclusive of poly(A), and contains two overlapping open reading frames (ORFs). The P + C mRNA encodes two proteins, the 602 amino acid nucleocapsid phosphoprotein P and the 199 amino acid non structural protein C. Peptide mapping confirmed that the two proteins are unrelated. Hybrid-arrest translation experiments assigned each of the two proteins to its respective ORF. These studies showed that the coding strategy of the PF3 P + C mRNA is similar to that of Sendai virus. Amino acid sequence alignment showed that the P and C proteins of PF3 and Sendai virus represent homologous pairs. However, these homologies are represented by high contents of accepted amino acid substitutions and by similarity in hydropathy profiles rather than by high contents of exact amino acid matches. Homology with the P and C proteins of measles, canine distemper and respiratory syncytial viruses was at the threshold of significance. The patterns of amino acid sequence homology among the paramyxovirus HN, F, NP, P and C proteins are compared. PMID- 3025347 TI - T cell-macrophage interactions in the immune response to herpes simplex virus: the significance of interferon-gamma. AB - The antiviral properties of a herpex simplex virus type 1-specific 'helper' T cell clone were investigated. The clone was found to be deficient in interleukin 2 production, although it produced interleukin 3 and interferon-gamma upon stimulation with the virus in vitro. Supernatants containing these lymphokines were observed to increase the virocidal activity of macrophages in vitro and furthermore induced these cells to mediate cytotoxic activity against virus infected target cells. Macrophage activation was linked to the presence of interferon-gamma in the clone supernatant. The implications of these results for protection against this virus in vivo are discussed. PMID- 3025348 TI - Comparison of the spike precursor sequences of coronavirus IBV strains M41 and 6/82 with that of IBV Beaudette. AB - The nucleotide sequences of the spike precursor genes of infectious bronchitis virus strains M41 and 6/82 have been determined and compared with that of the Beaudette strain which we have previously sequenced. The two Massachusetts strains, M41 and Beaudette, were found to be remarkably similar, having only 3.7% of the amino acids different. The situation with 6/82, one of the new field isolates, is quite different and this strain had 13.8% of its amino acids different from Beaudette. The differences identified are discussed in terms of the structural features of the spike protein. PMID- 3025349 TI - Nucleotide sequence of a cDNA clone of RNA segment 10 of bluetongue virus (serotype 10). AB - The complete sequence of the double-stranded RNA segment that codes for a non structural protein (P8) of bluetongue virus serotype 10 has been determined from a cDNA clone inserted into the plasmid pBR322. The segment 10 RNA of the virus (S10 RNA) is deduced to be 822 base pairs long (0.5 X 10(6) daltons) and has an open reading frame in one strand capable of coding for a protein with a calculated size of 25,572 daltons (229 amino acids) and a net charge of +5.5 at neutral pH. PMID- 3025350 TI - Rotavirus fecal IgA antibody response in adults challenged with human rotavirus. AB - Our studies of rotavirus challenge in adult volunteers enabled us to evaluate the relationship of pre-existing antirotavirus fecal IgA antibody to infection and illness and to investigate the local response to this infection. No relationship could be found between the pre-existing levels of fecal antirotavirus IgA antibody and protection from infection or illness. A greater than six-fold increase in the level of antibody was seen in 16/19 infected volunteers with determinable increases but in 0/15 controls who received less than the minimal infectious dose of rotavirus. Antibody levels increased rapidly in infected volunteers and were consistent with an anamnestic response. Two of seven volunteers who received an infectious dose of rotavirus but were considered uninfected on the basis of other laboratory methods had greater than or equal to six-fold increases of fecal antibody and one of these experienced symptoms compatible with a rotavirus infection. This finding indicates that an increase in fecal antibody may be a reliable indicator of rotavirus infection even in the absence of detectable shedding or seroconversion. PMID- 3025351 TI - Significance of specific Epstein-Barr virus IgA and elevated IgG antibodies to viral capsid antigens in nasopharyngeal carcinoma patients. AB - The feasibility of using elevated Epstein-Barr virus (EBV) specific-IgG antiviral capsid antigen (VCA) and IgA anti-VCA antibody levels as an aid in diagnosis of nasopharyngeal carcinoma (NPC) was analyzed by determination of serum antibody titers to EBV in 54 NPC patients, 114 healthy blood donors, and 40 family members by the immunoperoxidase assay (IPA). No significant difference was found in the prevalence rate of EBV IgG anti-VCA antibodies (titer greater than or equal to 20) between the patient group and the control and family groups (100% vs 92% and 90%, respectively). The prevalence rate of elevated EBV IgG anti-VCA titers (greater than or equal to 80, greater than or equal to 160, greater than or equal to 320, greater than or equal to 640) was significantly higher in the NPC patients than in controls. For example, at an IgG titer of greater than or equal to 320, the prevalence rate was 82% in the NPC patient group and 1.7% in the controls (P less than 0.0001). The prevalence of EBV IgA anti-VCA antibodies (greater than or equal to 10) was significantly higher in the NPC patients than in control and family groups (82% vs 6.1% and 0%, respectively). The prevalence rate for elevated EBV IgA anti-VCA (greater than or equal to 20) was found to be significantly higher (P less than 0.0001) in NPC patients than in the control group (70% vs. 1.7%). A significantly high proportion (P = 0.0004) of NPC patients who had serum EBV IgA anti-VCA titers of less than 20 had elevated IgG titers to VCA greater than or equal to 320 (21% vs 1.7% among controls). It appears that testing for IgG antibodies at a serum dilution of 1:320 and for IgA antibodies at a dilution of 1:20 by the IPA technique comprises the best combination for the differentiation between NPC patients and health controls (91% vs 3.4%), and it is suggested that these be used as screening markers for NPC patients. PMID- 3025352 TI - Visualization of replicating herpes simplex virus in cervical dorsal root ganglia of mice following explant of individual ganglion onto susceptible indicator cells. AB - Herpes simplex virus (HSV) establishes a latent state in the sensory ganglia of the peripheral nervous system of its natural or experimental host following primary infection. At various times thereafter, the virus can be reactivated from the latent state whereby it migrates back to the periphery and sometimes initiates a clinical syndrome referred to as recurrent disease. We inoculated mice in the right ear pinna and, following recovery from primary infection, killed the mice at various intervals following either the presence or absence of peripheral stimulations. Explanted cervical dorsal root ganglia yielded HSV in culture and was positive for HSV-like virus particles when viewed with the electron microscope. Hematoxylin and eosin staining showed neuron degeneration, and corresponding HSV-specific immunoperoxidase stains were also positive. The data indicate that ganglionic cells are capable of supporting replicating HSV and that, in vitro, numerous ganglionic cells can be infected simultaneously. PMID- 3025353 TI - Human cytomegalovirus replicates in gamma-irradiated fibroblasts. AB - Because of the unique interdependence of human cytomegalovirus (HCMV) and the physiological state of the host cell, we evaluated the ability of human foreskin fibroblasts (HFF), exposed to gamma radiation, to support HCMV growth. Irradiation of HFF with 2,500 rADS prevented cellular proliferation and suppressed cellular DNA, but not RNA or protein synthesis. Treatment of HFF cells with 2,500 rADS 6 or 48 hours prior to infection did not alter the time course or virus yield during HCMV replication. Virus plaquing efficiency in irradiated cells was comparable to that of nonirradiated cells. As judged by thymidine incorporation and BUdR inhibition of virus replication, HCMV infection induced both thymidine kinase activity and host cell DNA synthesis in irradiated cells. In addition, virus could be recovered from HFF exposed to radiation 0-2 days after infection with HCMV. These studies indicate that the damage to cells by gamma irradiation does not alter the capacity of host cells to support HCMV replication. PMID- 3025354 TI - Estimation of the B lymphocyte precursor frequencies to herpes simplex type 1 glycoproteins by a limiting dilution assay. AB - The precursor frequency of B lymphocytes from Balb/c mice producing HSV-1 glycoprotein B (gB), glycoprotein C (gC), and glycoprotein D (gD) antibody was determined by limiting dilution analysis under conditions to detect antibody from the clonal progeny of a single B cell precursor. In spleens of naive mice the average gC frequency was 1/48,917 +/- 5,550, while gD was 1/73,330 +/- 15,898, and gB frequency was in excess of 1/100,000. Immunization with live HSV-1 (KOS) increased the B cell frequencies of all three glycoproteins to approximately 1:3,000; however, the serum gB antibody ELISA titer was fivefold higher than gC or gD. PMID- 3025355 TI - Unusual monocyte-lymphocyte interactions determine the specificity of the immune specific interferon response induced by Newcastle disease and herpes simplex viruses. AB - Virus-induced immune-specific interferon (IS-IFN) is produced by previously sensitized peripheral blood mononuclear leukocytes (PBML) three to five days after they are placed in tissue culture. This IS-IFN is readily identified on the basis of its time of production and its synergistic composition of alpha and gamma interferons. The current studies demonstrate that circulating monocytes control the specificity and magnitude of the IS-IFN response. No IS-IFN is produced by PBML that are heavily depleted of monocytes. Immune-specific IFN production is enhanced in PBML cultures that are partially depleted of monocytes. Partial monocyte depletion permits virus to induce the production of IS-IFN by unsensitized PBML. PMID- 3025356 TI - Protective effect of preexisting rotavirus-specific immunoglobulin A against naturally acquired rotavirus infection in children. AB - The preexisting levels of rotavirus IgA and IgG were measured in 225 children aged 6 months to 7 years in November, ie, before the "rotavirus season" from January to April. During the following 6 months, all episodes of acute gastroenteritis (GE) were evaluated clinically according to a score system and feces was examined for rotavirus, pathogenic bacteria, and parasites. Furthermore, rotavirus GE (n = 45) as well as asymptomatic rotavirus infections (n = 29) were diagnosed serologically. The preexisting concentrations of rotavirus IgA and IgG measured by ELISA were similar in these two groups. However, preexisting rotavirus IgA in the group of children who developed rotavirus GE correlated with less severe symptoms. Thus vomiting was found in 24% and 63% of the children with detectable and undetectable rotavirus IgA, respectively (P less than 0.025). Moreover, according to the total symptom score of rotavirus GE, 52% of the children with detectable preexisting rotavirus IgA had mild symptoms compared with only 13% of those with undetectable concentrations (P less than 0.025). Rotavirus IgG did not have any protective effect. Age per se had a protective effect; older age (greater than 1.5 years) was related to mild symptoms. According to previous studies of local and intestinal antibody response to a rotavirus GE, it is suggested that rotavirus IgA in serum reflects the immunological status of the intestine concerning rotavirus. It is recommended that studies of rotavirus vaccines include rotavirus IgA response and its protective effect. PMID- 3025357 TI - Detection of varicella-zoster virus in lymphocytes by DNA hybridization. AB - The availability of cloned varicella-zoster virus (VZV) DNA probes allows rapid detection of viral-specific DNA by "dot-blot" hybridization in lymphocytes or in lesion aspirates. Thirty-six blood specimens were obtained from 25 patients with suspected varicella or zoster. VZV-specific DNA was demonstrated in 15 lymphocyte preparations of nine patients with varicella and in one with disseminated zoster out of five patients with zoster. VZV-specific DNA was detected prior to rise in antibodies, indicating early viremia in these patients. Virus isolation from lesions and serological tests confirmed VZV infections. VZV-specific DNA was detected in lymphocytes of three patients out of six with active herpetic lesions, whereas it was not detected in lymphocyte specimens from two patients with undiagnosed rash or four with lymphoproliferative diseases, who did not present varicella or zoster, or from 18 healthy controls. No signal was obtained in herpes simplex virus (HSV)-infected and -uninfected cell lines. The hybridization assay proved that specific and viral or cellular DNAs other than VZV did not cross-hybridize with the probe. The sensitivity limit of detection was 4-15 pg of homologous DNA, and the assay was accomplished within 72-96 hr. These results point to the possible rapid diagnosis of VZV infection in patients suspected of varicella or generalized zoster. In addition, simultaneous infection with both VZV and HSV seems to occur in some patients. PMID- 3025358 TI - Protection of Junin virus-infected marmosets by passive administration of immune serum: association with late neurologic signs. AB - Argentine hemorrhagic fever (Junin virus) is a human viral disease for which immune therapy proves effective, though a late neurologic syndrome is occasionally associated with the treatment. We attempted to determine in the infected marmoset Callithrix jacchus whether immune therapy leads to protection and/or CNS damage. Fifteen C jacchus were inoculated with 10(3) tissue culture infectious dose 50% (TCID50) of the XJ strain of Junin virus. On day 6 post infection (pi), 12 primates were treated with homologous immune serum. Animals were observed daily; and hematologic, serologic, virologic, and histologic studies were performed. All primates, both treated and controls, presented leukopenia, thrombocytopenia, anemia, and weight loss from day 14 pi onward. The three control animals died on days 22, 25, and 32 pi. Among the 12 treated monkeys, 3 died on days 21, 22, and 29. Hematologic values returned to normal during the second month; initial weight was recovered by the fourth month. Three out of the nine survivors showed neurologic alterations of various degrees, with hind-limb paralysis in the most severe case. Among treated monkeys, viremia and viral titers in the lungs, kidney, and lymph nodes were lower than in controls. Neutralizing antibodies were present in high titers in all treated marmosets, except in the one presenting paralysis in which values were minimal and viral persistence was detected in CNS. In conclusion, immune serum treatment of Junin virus-infected marmosets was found to reduce mortality from 100% to 25%. Viremia and viral titers in organs were lowered, and late neurologic signs appeared in 30% of treated survivors. PMID- 3025359 TI - Ethanolamine kinase activity in purified myelin of rat brain. AB - Highly purified rat brain myelin showed a significant level of ethanolamine kinase, amounting to 17% of the specific activity of whole brain homogenate. This kinase level in myelin was an order of magnitude higher than that of lactate dehydrogenase, a marker for cytosol. Subcellular distribution studies revealed that in addition to myelin, this kinase was present in the P1, P2, P3, and cytosolic fractions with highest relative specific activity in the latter. The possibility that myelin activity resulted from adsorption of the soluble enzyme was unlikely since activity was retained in myelin that had been washed with buffered sodium chloride or taurocholate. Mixing experiments and repeated purification further indicated that the enzyme is intrinsic to myelin. Kinetic studies indicated similar Km values for ethanolamine in the microsomal, cytosolic, and myelin fractions but a significantly lower apparent Km for ATP in myelin. This and other differences suggested the possible existence of isozymes. Establishment of the presence of this kinase completes the list of phospholipid synthesizing enzymes needed to synthesize phosphatidylethanolamine from diacylglycerol within the myelin membrane. PMID- 3025360 TI - Translocation of protein kinase C in anterior pituitary tumor cells. AB - Previous studies have shown that phorbol esters and lithium each stimulate the secretion of adrenocorticotropic hormone (ACTH) by the anterior pituitary tumor cell line AtT20/D16-16. Pretreatment with either lithium or phorbol ester desensitizes the cells to subsequent stimulation by phorbol ester. An early consequence of phorbol ester action in other systems is the translocation of protein kinase C from cytosol to membranes. We have assayed protein kinase C activity in cytosol and membranes of AtT20 cells after treatment with phorbol dibutyrate, lithium, or other agents that stimulate secretion of ACTH in these cells. Phorbol dibutyrate clearly induced translocation of protein kinase C, but lithium treatment did not cause translocation itself, nor did pretreatment with lithium affect the translocation induced by phorbol dibutyrate. These results are consistent with a role for translocation of protein kinase C in the stimulatory and desensitizing effects of phorbol esters but fail to implicate translocation in the actions of lithium on AtT20 cells. PMID- 3025361 TI - [3H]neurokinin B and 125I-Bolton Hunter eledoisin label identical tachykinin binding sites in the rat brain. AB - [3H]Neurokinin B ([3H]NKB) of high specific activity (75 Ci/mmol) was synthesized for study of its binding to crude synaptosomes from the rat cerebral cortex. The specific binding of [3H]NKB (75% of total binding) was temperature dependent, saturable, and reversible. Scatchard analyses and Hill plots showed the existence of a single population of noninteracting binding sites (KD = 4.3 nM; Bmax = 123 fmol/mg of protein). Competition studies indicated the following rank order of potencies among tachykinins: NKB greater than eledoisin (E) greater than kassinin greater than physalaemin greater than neurokinin A (NKA) greater than substance P (SP), a result suggesting that NKB might be the endogenous ligand for [3H]NKB binding sites. It is of interest that 127I-Bolton Hunter (BH) NKA (127I-BHNKA) was much more potent than NKA in inhibiting the specific binding of [3H]NKB, which raises certain questions concerning the use of 125I-BHNKA as a ligand for NKA binding sites in the brain. These results, as well as those obtained with different SP analogues, show a close similarity to those obtained previously with 125I-BHE binding to cortical synaptosomes. This suggested that the two ligands labeled identical binding sites. In addition, using either [3H]NKB or 125I-BHE as ligands, similar displacement curves were obtained with increasing concentrations of NKB and 127I-BHE. The similarity of the [3H]NKB and 125I-BHE binding sites was further confirmed by comparison of their localization on rat brain sections by autoradiography. The distribution of binding sites for [3H]NKB and 125I-BHE was identical throughout the brain, and the highest density of binding sites for the two ligands was found in layers IV and V of the cerebral cortex, the paraventricular nucleus of the hypothalamus (magnocellular part), and the ventral tegmental area. PMID- 3025362 TI - gamma-Aminobutyric acid-activated 36Cl- influx: a functional in vitro assay for CNS gamma-aminobutyric acid receptors of insects. AB - The effects of gamma-aminobutyric acid (GABA) on the uptake of 36Cl- into a membrane microsac preparation from isolated nerve cords of the cockroach Periplaneta americana was studied. On addition of 1 microM GABA (after 4-s incubation, then rapid quenching) the influx of 36Cl- was stimulated to a level 75% above that of the control value. This stimulation was reduced by picrotoxin (100 microM), but was not significantly affected by bicuculline (100 microM). Results of 36Cl- influx experiments are in agreement with data obtained from radiolabelled ligand binding assays and electrophysiological investigations on the same tissue. The method described represents a functional in vitro assay for CNS GABA receptors of insects. PMID- 3025363 TI - Purification and amino terminal sequencing of human melanoma nerve growth factor receptor. AB - The nerve growth factor (NGF) receptor, solubilized with Triton X-100 detergent, has been purified from human melanoma cell line A875. Purification to near homogeneity was achieved by chromatography on wheat germ agglutinin-agarose, followed by immunoaffinity chromatography on Sepharose columns coupled with anti NGF receptor monoclonal antibody (MAb). The purified receptor, a 75,000-dalton protein, retains the capacity to bind NGF as well as anti-receptor MAbs. Final purification was achieved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The sequence of amino acid residues at the amino terminus has been determined. Possible sequence homology between the NGF receptor and several other proteins is discussed. Using the purified receptor as immunogen, new MAbs to the NGF receptor have been produced. The NGF receptor was visualized by immunoperoxidase staining in tissue sections of dorsal root ganglia from monkeys. PMID- 3025365 TI - Stimulation-evoked Ca2+ fluxes in cultured bovine adrenal chromaffin cells are enhanced by forskolin. AB - Forskolin, 1 microM, increased acetylcholine (ACh)-stimulated 45Ca uptake by chromaffin cells. The stimulatory effects of forskolin decreased with increasing concentration of ACh. The attenuation of the effect of forskolin on 45Ca uptake as a function of ACh concentration correlated well with changes in the forskolin effect on ACh-evoked catecholamine (CA) release. Forskolin increased excess KCl- and veratrine-evoked CA release and 45Ca uptake. Forskolin by itself stimulated 45Ca efflux and enhanced ACh-, excess KCl-, and veratrine-stimulated 45Ca efflux. High doses of forskolin inhibited both ACh-evoked 45Ca uptake and CA release. The inhibitory action of forskolin was specific to receptor-mediated response because excess KCl- and veratrine-stimulated 45Ca uptake and CA release were not inhibited. Forskolin, 0.3-30 microM, dose-dependently increased caffeine stimulated CA release and 45Ca efflux in the absence of Ca2+ in the medium, and the effects were mimicked by dibutyryl cyclic AMP. These results suggest that cyclic AMP increases stimulation-induced CA release by enhancing calcium uptake across the plasma membrane and/or altering calcium flux in an intracellular calcium store. PMID- 3025364 TI - Enhancement of stimulation-evoked catecholamine release from cultured bovine adrenal chromaffin cells by forskolin. AB - Acetylcholine (ACh) increased cyclic AMP levels in cultured bovine chromaffin cells with a peak effect at 1 min after the addition. Pretreatment with forskolin (0.3 microM) enhanced the ACh-evoked cyclic AMP increase. The catecholamine (CA) release induced by ACh was enhanced by forskolin, but forskolin alone did not enhance the CA release. The effect of forskolin increased dose-dependently up to 1 microM, but decreased at higher concentrations. Dibutyryl cyclic AMP (DBcAMP) also enhanced ACh-evoked CA release, but the effect was less potent than that of forskolin. Forskolin enhanced both [3H]norepinephrine ([3H]NE) and endogenous CA release evoked by 30 mM K+ from cells that were preloaded with [3H]NE. The effects of forskolin were substantial when CA release was evoked with low concentrations of ACh or excess K+, but decreased with higher concentrations of the stimulants. Forskolin also enhanced the CA release induced by ionomycin and veratrine, or by caffeine in Ca2+-free medium. The potentiation by forskolin of the ACh-evoked CA release was manifest in low Ca2+ concentrations in the medium, but decreased when Ca2+ concentration was increased. These results suggest that cyclic AMP may play a role in the modulation of CA release from chromaffin cells. PMID- 3025366 TI - Stimulation of phosphoinositide hydrolysis by serotonin in C6 glioma cells. AB - 5-Hydroxytryptamine (serotonin or 5-HT) stimulated the incorporation of 32Pi into phosphatidylinositol (PI) but not into polyphosphoinositides in C6 glioma cells with an EC50 of 1.2 X 10(-7) M. The phosphoinositide response was blocked by the 5-HT2 antagonists ketanserin and spiperone but inhibited only partly by methysergide and mianserin. Atropine, prazosin, and yohimbine did not block the response, whereas fluphenazine and haloperidol did so partially but also inhibited basal incorporation by approximately 30%. The 5-HT1A agonist 8-hydroxy 2(di-n-propylamino)tetralin did not cause stimulation. Incubation with 5-HT (1 microM) for 1 h increased the incorporation of [2-3H]myoinositol into all phosphoinositides but not into inositol phosphates (IPs). Li+ alone at 10 mM increased labeling in inositol bisphosphate (IP2) and trisphosphate (IP3), whereas labeling in IP and phosphoinositides remained unaltered. Addition of 5-HT had no effect on this increase. Mn2+ at 1 mM enhanced labeling in PI, PI-4 phosphate, lyso-PI, glycerophosphoinositol, and IP, but the presence of 5-HT again did not cause further stimulation. 5-HT also stimulated the release of IPs in cells prelabeled with [2-3H]myo-inositol, incubated with LiCl (10 mM) and inositol (10 mM), and then exposed to 5-HT (1 microM). Radioactivity in IP2 and IP3 was very low, was stimulated approximately 50% as early as 30 s, and remained elevated for at least 20 min. Radioactivity in IP was at least 10 times as high as in IP3 but was increased only from 3 min on with a peak at 20 min, when the elevation was approximately 40 times that in IP3.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3025367 TI - Release of leukotriene B4 from sublethally injured oligodendrocytes by terminal complement complexes. AB - In the present study, the interaction of the terminal complement complexes with oligodendrocytes was investigated for observation of its effect on membrane lipid hydrolysis. [14C]Arachidonic acid was incorporated into the membrane lipids of cultured oligodendrocytes before sensitization with anti-galactocerebroside antiserum. Cells were then exposed to excess C6-deficient rabbit serum reconstituted with limiting doses of C6 to form various numbers of C5b-9 complexes. Qualitative analysis of the supernatants by HPLC revealed the presence of compounds that coeluted with arachidonic acid and its oxygenated derivatives, prostaglandin E2, leukotrienes E4 and B4, and 15-hydroxyeicosatetraenoic acid. The kinetics of leukotriene B4 release by excess C5b-8 was quantitated by radioimmunoassay. Leukotriene B4 release approached a maximum around 30 min, and C6 dose-response studies performed at 1 h showed that maximal levels of leukotriene B4 were detected over a range of sublytic C5b-9 attack. Maximal release of leukotriene B4 was also achieved by C5b-8 without further enhancement by addition of lytic doses of C9. Results indicate that sublytic attack of oligodendrocytes by complement induces release of lipid-derived inflammatory mediators. PMID- 3025368 TI - Degradation of luteinizing hormone-releasing hormone by neuroblastoma cells and their membrane: evidence for the involvement of a thiol protease and angiotensin converting enzyme. AB - Luteinizing hormone-releasing hormone was degraded by cells of the N-18 line of mouse neuroblastoma and their membrane. Cleavage products were separated by HPLC and identified by amino acid analysis. Fragments (1-3), (4-5), and (6-10) were major cleavage products. All the products increased in level as a function of time except for fragment (1-5), which increased in amount only during a short incubation time and then decreased. The accumulation of fragment (1-5) was increased in the presence of captopril or EDTA, whereas that of fragments (1-3) and (4-5) decreased inversely. On the other hand, the generation of either fragment (1-3) or (4-5) was stimulated by the presence of Cl-. The results suggest that the conversion of fragment (1-5) into fragments (1-3) and (4-5) is catalyzed by angiotensin-converting enzyme. p-Chloromercuribenzoate inhibited the formation of fragment (1-5), a result suggesting the involvement of a thiol protease in this formation. Thus, the degradation of luteinizing hormone releasing hormone by neuroblastoma cells and their membrane seems to take place mainly through the cleavage of the Tyr5-Gly6 bond by a thiol protease, followed by the release of the dipeptide Ser-Tyr from the liberated fragment (1-5) by angiotensin-converting enzyme. It is further suggested that the thiol protease and angiotensin-converting enzyme are also responsible for the initial minor cleavages of the Gly6-Leu7 bond and the Trp3-Ser4 bond, respectively. PMID- 3025369 TI - Photoaffinity labeling of benzodiazepine receptor proteins with the partial inverse agonist [3H]Ro 15-4513: a biochemical and autoradiographic study. AB - Photolabeling of the benzodiazepine receptor, which to date has been done with benzodiazepine agonists such as flunitrazepam, can also be achieved with Ro 15 4513, a partial inverse agonist of the benzodiazepine receptor. [3H]Ro 15-4513 specifically and irreversibly labeled a protein with an apparent molecular weight of 51,000 (P51) in cerebellum and at least two proteins with apparent molecular weights of 51,000 (P51) and 55,000 (P55) in hippocampus. Photolabeling was inhibited by 10 microM diazepam but not by 10 microM Ro 5-4864. The BZ1 receptor selective ligands CL 218872 and beta-carboline-3-carboxylate ethyl ester preferentially inhibited irreversible binding of [3H]Ro 15-4513 to protein P51. Not only these biochemical results but also the distribution and density of [3H]Ro 15-4513 binding sites in rat brain sections were similar to the findings with [3H]flunitrazepam. Thus, the binding sites for agonists and inverse agonists appear to be located on the same proteins. In contrast, whereas [3H]flunitrazepam is known to label only 25% of the benzodiazepine binding sites in brain membranes, all binding sites are photolabeled by [3H]Ro 15-4513. Thus, all benzodiazepine receptor sites are associated with photolabeled proteins with apparent molecular weights of 51,000 and/or 55,000. In cerebellum, an additional protein (MW 57,000) unrelated to the benzodiazepine receptor was labeled by [3H]Ro 15-4513 but not by [3H]flunitrazepam. In brain sections, this component contributed to higher labeling by [3H]Ro 15-4513 in the granular than the molecular layer. PMID- 3025370 TI - Localization of the phosphorylation sites for different kinases in the microtubule-associated protein MAP2. AB - The phosphorylation of microtubule-associated protein 2 (MAP2) by four different kinases was studied in vitro to determine whether MAP2 is phosphorylated in its tubulin binding region or in the microtubule projection portion. Fragments corresponding to both regions of MAP2 were produced not only by chymotrypsin or trypsin digestion, but also using pepsin, a broad chain-specificity protease, a result supporting previous notions of the two-domain structure of MAP2. The position of these two functional domains was determined with respect to the carboxy terminal of the molecule, by labeling MAP2 exclusively at the carboxy terminal and subjecting it to pepsin digestion. The results suggested that the projection region of MAP2 contained the carboxy terminal of the protein. A phosphorylation map was constructed by subjecting phosphorylated MAP2 to enzymatic digestion using Staphylococcus aureus V8 protease or to chemical cleavage using N-chlorosuccinimide. The results indicated that all four kinases phosphorylated MAP2 in a 42-kilodalton peptide that contained the tubulin binding region but differed in the level and localization of the sites at which they phosphorylated the projection of MAP2. PMID- 3025371 TI - Regulation of dipeptidyl aminopeptidase I and angiotensin converting enzyme activities in cultured murine brain cells by cortisol and thyroid hormone. AB - Cultures of dissociated brain cells from 15-day-old fetal mice were grown in the presence and absence of 20 or 50 nM triiodothyronine (T3), 30 or 300 nM cortisol, and 30 nM cortisol plus 50 nM T3 added to chemically defined media or in media supplemented with 15% serum from control and hypothyroid calves. The specific activities of five lysosomal enzymes--N-acetyl galactosaminidase, beta glucuronidase, beta-galactosidase, cathepsin B, and dipeptidyl aminopeptidase I (DAP-I)--were higher in cells grown in calf serum than in cells grown in defined media. Of these enzymes, only DAP-I was elevated in activity when the cells were grown in hypothyroid calf serum instead of control calf serum. Elevation of DAP-I activity was reversed by addition of 20 nM T3 to hypothyroid calf serum. The enzymatic properties of DAP-I were similar whether the cells were grown in control or hypothyroid calf serum and were similar to those reported for human fibroblasts and the purified enzyme. When the cells were grown in defined media, cortisol decreased the activities of all lysosomal enzymes, with 300 nM cortisol being more effective than 30 nM cortisol. Addition of 50 nM T3 to 30 nM cortisol decreased DAP-I activity more than 30 nM cortisol alone, but 50 nM T3 alone in defined media did not alter DAP-I levels. The reduction of DAP-I activity in these cells by T3 required cortisol, unidentified components in serum, or both.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3025372 TI - Dissociation of the stellate morphology from intracellular cyclic AMP levels in cultured rat brain astroglial cells: effects of ganglioside GM1 and lysophosphatidylserine. AB - Secondary microcultures of newborn rat cerebrum astroglial (AG) cells, maintained in a serum-free, chemically defined medium, were treated with various agents known to elevate intracellular cyclic AMP (cAMP) levels. Earlier studies had shown these drugs to induce a process-bearing (stellate) morphology in the AG cells, a response that was antagonized by the presence of gangliosides. One millimolar dibutyryl cyclic AMP (dBcAMP), 10 microM forskolin, 12 nM cholera toxin, and 30 microM isoproterenol all raised intracellular cAMP levels, from basal values of 3 pmol/10(6) cells to 30-30,000 pmol/10(6) cells, depending on the agent tested. dBcAMP caused the greatest elevation, and forskolin the least. The timing and/or the level of the AMP response did not precisely correlate with those of the stellation response. Values of ED50 with the four agents, as determined for the cAMP response, were always higher than stellation ED50 values in all treatments, and ED50 did not correlate with the maximal levels of cyclic AMP induced by the four agents. The capacity of ganglioside GM1 to block the stellation response to the four agents was not accompanied by a similar capacity to block the cAMP responses. Lysophosphatidylserine (lysoPS) had the capacity to induce AG cell stellation as well, without altering the basal level of cAMP. Both lysoPS and gangliosides, therefore, may act directly on the cellular machinery underlying the stellation response without involving changes in intracellular AMP. PMID- 3025373 TI - Proenkephalin B messenger RNA in porcine tissues: characterization, quantification, and correlation with opioid peptides. AB - Concentrations of proenkephalin B (PENK B) mRNA in porcine brain, pituitary, spinal cord, and peripheral tissues were measured using RNA blotting and solution hybridization. A single hybridizing species of approximately 2,800 bases in size was present in the CNS, with the highest concentration in the caudate nucleus, followed by hypothalamus and hippocampus. The abundance of PENK B mRNA ranged from 22 pg/micrograms of poly(A)-rich RNA in caudate nucleus to less than 0.1 pg/microgram in cerebellum. Concentrations of immunoreactive PENK B-derived peptides showed a similar distribution, with the exception of the hypothalamus, which had lower PENK B mRNA levels than expected from peptide concentrations. PENK B mRNA of the same size as in the brain was also found in the anterior lobe of the pituitary and in the heart ventricle, whereas in intestine, lung, and kidney, smaller mRNA species of 1,800 bases became apparent by RNA blot analysis. An intermediate size of 2,200 bases was found in heart atrium. As revealed by S1 mapping, however, these smaller mRNAs are not completely homologous with PENK B mRNA, but rather may represent closely related mRNAs from a different gene(s). PMID- 3025374 TI - The rigid spine syndrome. AB - Four patients are reported, 3 females and 1 male, with (as a prominent symptom of muscle disease) limitation of flexion of cervical and dorsolumbar spine. The nosological classification of these cases is discussed. In two patients there was evidence of an inclusion body myositis. At necropsy one of these patients had a remarkable distribution of muscle changes. PMID- 3025375 TI - Regional cerebral blood flow in patients with Parkinson's disease under chronic levodopa therapy: measurements during "on" and "off" response fluctuations. AB - In ten Parkinsonians who developed dose-related response fluctuations under long term levodopa therapy, regional cerebral blood flow (rCBF) and plasma levodopa levels were measured simultaneously, once during an "off" phase and again at an "on" stage of clinical benefit induced by a single oral dose of levodopa. Although plasma levodopa increased threefold during the "on" period, rCBF and the degree of its reduction from normal age-matched control values remained unchanged and similar to those in the "off" phase. Study suggests that the rCBF decreases in Parkinson's disease are unaffected by levodopa and are not due to deficient dopaminergic neurotransmission in the brain. PMID- 3025376 TI - The clinical spectrum of inflammatory-angiopathic neuropathy. AB - Thirty-three patients with inflammatory-angiopathic neuropathy diagnosed by sural nerve biopsy, were investigated to determine the underlying disease. Twenty-six patients had symmetrical sensorimotor polyneuropathy and seven had mononeuropathy multiplex. An aetiology for inflammatory-angiopathic neuropathy was found in only eight patients: typical collagen vascular disease in five and malignant tumour in three. Sixteen patients received prednisone and/or immunosuppressive drug therapy and 12 (75%) responded to treatment. This study demonstrates that typical collagen vascular diseases are not the most common cause of inflammatory angiopathic neuropathy, that symmetrical polyneuropathy is seen in 75.8% of inflammatory-angiopathic neuropathy patients. Prednisone and/or immunosuppressive agents appear effective regardless of aetiology. PMID- 3025377 TI - Neuromuscular transmission in the murine mutants "motor end-plate disease" and "jolting". AB - Mice with the inherited disorder "motor end-plate disease" suffered from a progressive neuromuscular weakness and muscular wasting. The weakness resulted from a failure of evoked transmitter release from the motor nerve terminals. The failure in transmission was all-or-nothing in nature. The numbers of muscle fibres in skeletal muscle and myelinated axons in several major nerve trunks were no different from normal. The loss in muscle bulk was caused by the neuromuscular defect and not from a loss of motor units or muscle fibres. The inherited murine disorder "jolting" was allelic with "motor end-plate disease". Affected "jolting" mice suffered no detectable morphological abnormality in skeletal muscle or peripheral nerve. The physiological properties of skeletal muscle and the characteristics of neuromuscular transmission were indistinguishable from normal. PMID- 3025378 TI - Serum protein masking of the thermal sensitivity of the antiviral activity of purified human beta interferon: implications for clinical studies. AB - Human beta-interferon (HuIFN-beta) exhibits antiproliferative and antiviral properties. Successful clinical application of this drug depends on knowledge of the thermal stability of these activities under physiological conditions. In the present study, both the antiproliferative and antiviral activities were stabilized by the addition of very small quantities of serum proteins. This supplement was sufficient to mask the slightly higher thermosensitivity of the antiviral activity. In the absence of serum proteins, the values of both the half life and the energy of activation were higher for the antiproliferative activity than for the antiviral function. Each had a half-life of at least 24 h and identical values for the energy of activation in the presence of proteins furnished by 1% fetal bovine serum. This study provides additional evidence to support a thesis recently advanced by Carter et al. that the antiproliferative domain of glycosylated beta interferon may be separable from the antiviral domain. It is concluded that the efficacy of HuIFN-beta, under clinical conditions, will not be seriously impaired by thermal inactivation. Antiviral assays of serum may be freely substituted for antiproliferative assays during pharmacological studies. PMID- 3025379 TI - Murine cytomegalovirus infection of reaggregate cultures of fetal mouse brain. AB - We used cultures of reaggregate embryonic mouse brain cells to study murine cytomegalovirus (MCMV) infection of neural tissues. After 21 to 28 days in culture, aggregates were infected with MCMV and studied sequentially for 14 days using virus assay, electron microscopy and indirect immunofluorescence. Infectious virus could be recovered from aggregate cultures beginning three days after infection, and peak virus titers were observed on day 7 in aggregate tissues and on day 14 in culture fluids. By transmission electron microscopy, intranuclear viral nucleocapsids were identified in neural cells at the periphery of the aggregates on day 3. Infection then spread centripetally into aggregate tissues so that by day 14 the majority of neural cells contained intranuclear inclusions, numerous nucleocapsids, and mature virus particles. Virion production in neural cells was the result of a sequence of events that included budding of nucleocapsids from the nucleus and envelopment of cytoplasmic virus particles by membranes of the Golgi apparatus. These studies indicate that MCMV infection of murine brain aggregate cultures is a potentially useful in vitro system for the study of CMV infections of neural tissue. PMID- 3025380 TI - Quantitative characterization of ceruleospinal inhibition of nociceptive transmission in the rat. AB - A total of 51 dorsal horn units responsive to heat were isolated and their receptive fields characterized (i.e., response properties and adequate stimuli determined) in pentobarbital-anesthetized, paralyzed rats. In 39 of the 51 units, the descending inhibition of heat-evoked activity produced by focal electrical stimulation in the locus ceruleus/subceruleus (LC/SC) was examined. All units studied responded to mechanical stimulation, to electrical stimulation of the ipsilateral tibial nerve at intensities supramaximal to activate A-alpha, delta- and C-fibers, and to noxious heating (50 degrees C) of the footpad. The cutaneous receptive fields of all units were confined to the glabrous skin of the toes and footpad. All neurons examined were located in the dorsal horn of the spinal cord in laminae I-VI. Tracking experiments established that inhibition of heat-evoked dorsal horn unit activity could be reliably produced by focal electrical stimulation in both the contralateral and ipsilateral LC/SC. The inhibition produced by electrical stimulation in the LC/SC was intensity-, pulse duration-, and frequency-dependent. In six experiments, the efficacy of LC/SC stimulation produced inhibition of heat-evoked activity was compared using two pulse durations (100 and 400 microseconds); greater inhibition of heat-evoked activity was produced at lower intensities of stimulation at the 400-microseconds pulse duration. In 10 experiments, the frequency of stimulation was varied (25-200 Hz); stimulation at a frequency of 100 Hz resulted in maximal inhibition of heat evoked activity for stimulation sites both inside (n = 7) and outside (n = 3) the LC/SC. Inhibition of heat-evoked dorsal horn unit activity could be reliably produced by focal electrical stimulation in sites inside the LC/SC (n = 18). Significant descending inhibition of noxious heat-evoked spinal neuronal activity could also be produced by stimulation in pontine sites located outside the LC/SC, however, not as reliably. Systematic electrode tracks were made through the pons, using a searching stimulus of 100 microA, to locate sites medial, lateral, and ventral to the LC/SC from which significant descending inhibition could be produced. Stimulation in 156 sites outside the LC/SC at 100 microA produced inhibition of heat-evoked spinal unit activity to 50% of control or less in only 37 sites. Descending inhibition was characterized quantitatively from 14 of these 37 sites; the mean intensities of stimulation to inhibit heat-evoked activity to 50% of control were experimentally determined, and the mean thresholds of stimulation for inhibition and the mean recruitment indices were calculated.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3025381 TI - Suppression of the membrane defect by divalent cations in the Drosophila mutant shibire. AB - The single-gene mutant shibire (shits) is temperature-sensitive. It causes reversible paralysis at heat pulses greater than 29 degrees C by blocking synaptic transmission. The synapses of these heat-pulsed flies are depleted in vesicles but contain numerous cisternae. We report that such alterations in synaptic physiology and morphology of heat-pulsed flies can be suppressed by internal perfusion of salines with high concentrations (10-18 mM) of the divalent cations Ca2+ or Mg2+. Synaptic morphology in these perfused flies remains normal even when exposed to nonpermissive temperatures (greater than 29 degrees C); in addition, synaptic transmission maintains a high resistance to failure both in frequency and stimulus duration. We also observed many cisternae in close association with extrajunctional as well as with postsynaptic regions of the sarcolemma in heat-pulsed shits flies not perfused with the increased concentrations of divalent cations. Flies perfused with increased amounts of divalent cations lacked such cisternae in the sarcolemma. The evidence suggests that the divalent cations can mitigate an overall membrane defect expressed by the shits gene, perhaps by influencing lipid-phase transition behavior. PMID- 3025382 TI - Regulation of serotonin-stimulated phosphoinositide hydrolysis: relation to the serotonin 5-HT-2 binding site. AB - The hypothesis that serotonin (5-HT)-stimulated phosphoinositide hydrolysis is mediated by the 5-HT-2 binding site in cerebral cortex was tested by comparing antagonist Kd values determined by Schild analyses with Ki values at the 5-HT-2 binding site. A significant correlation was found between Kd values and Ki values at competing for 3H-ketanserin binding (R = 0.98), suggesting that the phosphoinositide-linked receptor and the 5-HT-2 site are identical. The 5-HT-2 mediated phosphoinositide response was then used as a measure of 5-HT-2 receptor sensitivity after in vivo treatments that alter the availability of 5-HT. Chronic treatment with the 5-HT-2 antagonist mianserin resulted in a significant decrease (52%) in the density of 5-HT-2 binding sites and a significant decrease (49%) in the maximal phosphoinositide hydrolysis response to 5-HT. Depletion of 5-HT levels with para-chlorophenylalanine or chemical denervation of serotonergic neurons with 5,7-dihydroxytryptamine had no significant effect upon 5-HT-2 receptor density or upon the phosphoinositide response to 5-HT. These data suggest that changes or lack of changes in 5-HT-2 receptor density following in vivo manipulations reflect the functional state of the receptor. PMID- 3025383 TI - Cross-linking of atrial natriuretic peptide to binding sites in rat olfactory bulb membranes. AB - Binding sites for 125I-atrial natriuretic peptide (ANP)2 in rat olfactory bulb membranes have been studied using pharmacological and biochemical methods. Various unlabeled ANP-related peptides were tested for the ability to inhibit the binding of the radioligand in membrane binding assays. ANP(92-126) and ANP(99 126) were the most potent inhibitors tested, both exhibiting an IC50 value of 0.40 nM. ANP(103-126) and ANP(103-123) were 3 and 70 times less potent, respectively. ANP(111-126) was unable to inhibit the binding of the radioligand at a concentration of 1 microM. Several peptides unrelated to ANP were unable to inhibit the binding of the radioligand to rat olfactory bulb membranes. Membranes labeled with 125I-ANP were incubated with cross-linking agents and subjected to SDS-PAGE followed by autoradiography. A band possessing an apparent molecular mass of 116 kDa was identified. The labeling of this band was progressively decreased by increasing concentrations of unlabeled ANP(99-126) (IC50 = 0.6 nM) and by several other ANP-related peptides at nanomolar concentrations. For comparison purposes, ANP binding sites in rat aorta membranes were labeled with 125I-ANP and cross-linked using identical techniques. Three bands possessing molecular masses of 120, 72, and 62 kDa were identified. These results indicate that the ANP binding site in rat olfactory bulb membranes displays pharmacological and biochemical properties similar to peripheral ANP receptors. PMID- 3025384 TI - Hodgkin's disease with hypercalcemia detected by thallium-201 scintigraphy. AB - A 53-yr-old man with hypercalcemia was referred after an unsuccessful operative attempt to find a parathyroid adenoma. Metabolic evaluation showed relatively suppressed levels of parathyroid hormone with an elevation of serum 1,25 dihydroxyvitamin D. Thallium-technetium dual isotope imaging revealed localized mediastinal thallium uptake. A vascular mediastinal lesion was then demonstrated by arteriography, with subsequent surgical removal of a mass that proved to be lymphocyte predominant Hodgkin's disease. This case is noteworthy for the finding of isolated lymphocyte predominant Hodgkin's disease in the chest, the association of elevated serum 1,25-dihydroxyvitamin D with hypercalcemia that resolved postoperatively, and the uptake of thallium by the tumor. PMID- 3025385 TI - Radionuclide imaging of sequential torsions of the appendix testis. AB - Radionuclide imaging is the diagnostic procedure of choice for evaluation of acute disorders of the scrotum, permitting rapid differentiation of torsion of the spermatic cord from inflammatory processes. The method is less successful in diagnosing torsion of the testicular appendages. In the majority of reported cases of torsion of the appendix testis the scan appears normal. In a very limited number of cases increased perfusion to the affected side has been noted. We report a case of metachronous torsion of both the appendices testis. In each instance, there was decreased tracer uptake on the affected side, suggestive of early torsion of the testicle. The sequential involvement of each testis with a nine month interval between respective torsions provides a unique example where each testis serves, sequentially, as a normal control for the torsed contralateral appendage. PMID- 3025386 TI - Myocardial damage delineated by indium-111 antimyosin Fab and technetium-99m pyrophosphate. AB - Technetium-99m-labeled pyrophosphate and radiolabeled antimyosin antibodies are two infarct localizing agents with apparently different kinetics of localization. To determine whether these agents localize in a similar fashion in early acute myocardial necrosis, we studied the simultaneous distribution of 111In-labeled antimyosin and 99mTc-labeled pyrophosphate in dogs after intracoronary (i.c.) (n = 9) or intravenous (i.v.) (n = 9) administration of a mixture of these two agents in a reperfused infarct model. The mean infarct size (+/- s.d.) delineated by pyrophosphate [20.2 +/- 14.1 (i.v.), 29.8 +/- 12.3 (i.c.)] was larger than that by antimyosin [14.2 +/- 11.3 (i.v.) (p = 0.05), 20.0 +/- 11.8 (i.c.) (p = 0.05)] which was larger than that by triphenyl tetrazolium chloride [13.9 +/- 8.0 (i.v.) (p = 0.05), 15.3 +/- 6.5 (i.c.) (p = 0.05)]. This overestimation persisted whether the radiopharmaceuticals were administered by intracoronary or intravenous injections, although the latter with antimyosin was only slightly larger (TTC:AM = 13.9:14.2) (p = n.s.). There was a good correlation, however, between antimyosin and pyrophosphate delineated infarct sizes in dogs with intracoronary injection (y = 0.82x + 13.33, r = 0.79) or i.v. injection (y = 1.208x + 3.01, r = 0.97) of the mixture of the two agents. Since the images of the 111In and 99mTc activities were obtained consecutively by identical methods, the overestimation of infarct size by pyrophosphate cannot be due to differences in spatial resolution of the techniques used. The differences in the areas of myocardial damage delineated by pyrophosphate and antimyosin in our study most probably denote the area of viable but compromised myocardium. PMID- 3025387 TI - Standardization of nuclear medicine methods in cardiology. PMID- 3025389 TI - Multicentric reticulohistiocytosis accompanied by oral and temporomandibular joint manifestations. AB - A case of multicentric reticulohistiocytosis in a 56-year-old man with oral and temporomandibular joint manifestations is presented, which demonstrates typical clinical features of skin and mucosal nodules, and polyarthritis. PMID- 3025388 TI - Apparent small intestinal absorption of nitrogen and minerals from soy and meat protein-based diets. A study on human ileostomy subjects. AB - The apparent absorption of nitrogen and minerals was studied in 8 ileostomy subjects. Four different test diets containing 60 g of meat, rice and bread protein, or a 25% replacement of the protein with soy flour, soy concentrate or soy isolate, were randomly assigned to the subjects in 2-d periods. All animal protein was replaced by soy isolate for a fifth 2-d period in two of the subjects. Ileostomy contents were collected in 2-h intervals during the day and in one portion during night and immediately deep-frozen. The fiber components and the phytic acid in the diet were almost completely recovered in the ileostomy contents, whereas unabsorbed starch was less than 2% of the intake. A significantly lower protein digestibility was observed when the diets containing soy protein were fed. No difference in protein digestibility was found between the different soy protein products. A 25% replacement by soy protein had no obvious effect on apparent mineral absorption. A low protein digestibility was also observed when soy was the main source of protein, and a negative apparent absorption of zinc was found in both subjects. Although 25% of soy protein in the diet does not seem to impair mineral absorption significantly, small intestinal net absorption of nitrogen is less from the soy diets than from the meat diet. PMID- 3025390 TI - A method to quantify the hydroxylapatite needed for ridge augmentation. PMID- 3025392 TI - Determination of brain death. Ad Hoc Committee on Brain Death. PMID- 3025391 TI - Characterization of two cell lines, derived from two malignant fibrous histiocytomas. AB - We have investigated the phenotype and ultrastructure of tumour cells from two cell lines each derived from a malignant fibrous histiocytoma (MFH) as a means of studying the histogenesis of this group of tumours. The first MFH (MFH-I) was of the pleomorphic subtype, with a predominantly histiocytic appearance, the second was of the pleomorphic subtype associated with myxoid and storiform areas (MFH II). In vitro tumour cells from both neoplasms showed aberrant growth properties. Xenografts in nude mice from both neoplasms showed a similar histology to that of the original tumour. Both tumours showed hyaluronidase sensitive alcian blue staining. Phenotypic studies of the two cell lines and of the tumour tissues demonstrated that the cells differed in the presence of collagen types I and III. They did not show evidence of histiocytic, endothelial, leiomyoblastic, rhabdomyoblastic, lipoblastic of schwannian origin. Ultrastructurally, the two cell lines were found to be different. In vitro and in xenografts the cell type of MFH-I resembled a primitive mesenchymal cell. Whereas that of MFH-II resembled a fibroblast-like cell. We concluded that the group of MFH is heterogeneous and is probably derived from more than one progenitor cell. PMID- 3025393 TI - Live attenuated varicella vaccine. PMID- 3025394 TI - Acquisition of cytomegalovirus infection from birth to 10 years: a longitudinal serologic study. AB - We performed serial serologic tests for cytomegalovirus (CMV) antibody in 177 children born to low- and middle-income families in Houston from 1975 to 1983. Mean duration of participation in the study was 4.8 years (range 1 to 9.6 years). Most rapid acquisition of antibody occurred during the first and second years of life, 13.6% and 12%, respectively; thereafter, annual acquisition varied from 1.5% to 4.6%, up to 10 years. Overall, 59 (33%) of the group were known to seroconvert by age 10 years. This was a minimal figure because of loss to follow up. Analysis by the Kaplan-Meier method indicated that the probability of remaining seronegative was 65% at age 6 years, and 58% at age 8 years. Variables positively related to seroconversion by multivariate analysis were order of birth, seroconversion in a family member, and breast-feeding. During the first year of life, acquisition of CMV antibody was related to the seroimmune status of the mother. The variables of socioeconomic status, race, age of the mother, and attendance in a day care center did not appear to be related to seroconversion in these children. PMID- 3025395 TI - Human parvovirus infection in patients with sickle cell disease with and without hypoplastic crisis. PMID- 3025396 TI - Diagnostic red blood cell scintigraphy in GI tract bleeding from an intestinal hemangioma. AB - 99mTc-labeled red blood cell (RBC) scintigraphy in vivo established the diagnosis of intestinal hemangioma in a 6-year-old girl suffering from gastrointestinal (GI) tract bleeding who had undergone an operation for removal of hemangioma at the nail bed. Other examinations, double-contrast barium meal study and angiography, were unsuccessful in making the diagnosis. Intestinal hemangioma is rare in childhood, but a useful technique such as RBC scintigraphy should be performed to find the bleeding point of the GI tract, especially in case the patient has another obvious hemangioma. PMID- 3025397 TI - Childhood obesity: plasma potassium and Na-K-ATPase indicate that the electrolyte content of food may be important. PMID- 3025399 TI - Multiple duplications of the small intestine. AB - A case of coexistent cystic and tubular duplications of the small intestine is presented. Staged resection resulted in maximal preservation of small intestinal length. Intraoperative radionuclide scanning is suggested as a technique to assure complete removal of ectopic gastric mucosa. The literature regarding multiple duplications is reviewed and the management of duplications involving long segments of the small intestine is discussed. PMID- 3025398 TI - Chronic intussusception due to antral myoepithelioma. AB - Intussusception was seen on abdominal sonography and computed tomography in a 15 year-old boy who presented with a 6-week history of weight loss, vomiting, abdominal pain, abdominal mass, and hyperamylasemia. Laparotomy revealed a chronic gastroduodenal intussusception, the lead point of which was an antral myoepithelioma, a rare entity in this age group. PMID- 3025400 TI - Resection of a vipoma of the pancreas in a 15-year-old girl. AB - A 15-year-old girl had massive watery and protein-losing diarrhea. She was found to have a pancreatic islet cell tumor secreting high levels of vasoactive intestinal polypeptide. Eighty-five percent pancreatic resection was needed to remove the tumor. She has normal serum VIP levels and no tumor recurrence 6 years later. PMID- 3025401 TI - Congenital Wilms' tumor associated with consumption coagulopathy and hyperbilirubinemia. AB - A case of a term newborn infant with a congenital Wilm's tumor who developed severe jaundice and consumption coagulopathy is reported. After the removal of the tumor, the clotting factors became normal and the jaundice resolved. It is suggested that there was an association between the presence of the tumor and hemolysis and consumption coagulopathy. PMID- 3025402 TI - Bilateral Wilms' tumor. AB - Recent studies indicate a favorable outcome with bilateral Wilms' tumor. From 1971 to 1985, ten children between the ages of 6 months and 5 years were treated for this disease. During the early part of the series, five patients had nephrectomy on one side and partial nephrectomy on the other side, and one patient had bilateral partial nephrectomies at the same operation. More recently four patients initially had a biopsy of both tumors and lymph node sampling followed by chemotherapy. At the second-look procedure two patients had multiple biopsies because there was no obvious tumor. Histologically there was no tumor on the third-look procedure in these two patients. Nine patients had a favorable histology of the Wilms' tumor. One patient had a favorable histology on one side and an unfavorable type of histology on the other side. Eight patients are surviving between 6 months and 13 years. Two died of extensive disease within 16 months of diagnosis. One patient had an unfavorable histology. The good results following partial nephrectomies have led us to attempt to conserve additional tissue, as has been done in the last four patients. Our early results suggest biopsy of the tumor followed by chemotherapy, then a second look, and if necessary, third-look procedures may result in preservation of functioning renal tissue. PMID- 3025403 TI - Collagenase and collagenase inhibitor activities in crevicular fluid of patients receiving treatment for localized juvenile periodontitis. PMID- 3025404 TI - An unusual cause of bilateral mental neuropathy in an AIDS patient. Report of a case. AB - A case is reported of a patient with sudden onset, generalized toothache accompanied with a numb chin and lower lip. A thorough oral examination was negative. A complete medical evaluation revealed a positive HTLV-III antibody titer and acute lymphoblastic leukemia (Burkitt's-type). An unexplained trigeminal neuropathy should prompt the dentist to refer the patient for complete medical evaluation. A high index of suspicion of a malignant process should be maintained in all cases of unexplained numb lower lip and/or chin. PMID- 3025405 TI - Bioceramic implants in surgical periodontal defects. A comparison study. AB - The healing response of two commercially available bioceramics, beta tricalcium phosphate (TCP) and hydroxylapatite (HA), was compared after implantation in surgically created defects in dogs. Three 18 to 24 month old female Labrador dogs were used. Under general anesthesia 3-wall defects were created on the canines and premolars. The roots were planed, and reference notches were placed to identify the alveolar crest and the apical extent of the defects. TCP or HA was placed in alternating canine defects. The premolars received no implants and served as controls. Plaque management was accomplished by biweekly brushing with 0.2% chlorhexidine. Following healing periods of 5, 12 and 16 weeks, the dogs were sacrificed and perfused with 10% formalin. Six-mu step serial sections were evaluated by light microscopy. Healing against the root planed surface varied from a long junctional epithelium to a connective tissue reattachment in new cementum. TCP particles were actively resorbed by giant cells and macrophages and were incorporated into new bone matrix. The HA particles were encapsulated by fibrous connective tissue and rarely seen in contact with repairing bone. Bone formation was slower around HA particles at all time periods. Some evidence of HA particle resorption was seen at each time period. PMID- 3025406 TI - Structure-activity relationship of cardiac steroids having a doubly linked sugar and related compounds for the inhibition of Na+,K+-adenosine triphosphatase. AB - Forty-three different cardiac steroids having a doubly linked sugar and related compounds were tested for inhibition of Na+,K+-adenosine triphosphatase from guinea pig heart, and the structure-activity relationship has been discussed. The doubly linked glycosides showed higher activities than the respective genins. The inhibitory activities of these compounds were also dependent upon the presence of ring C substituents. The bufadienolide rhamnosides exhibited greater than three times the inhibitory activity of the parent genin, while the glucosyl residue exerted no significant activity. PMID- 3025407 TI - The action of mitomycin C on the induction of cyclic AMP phosphodiesterase and the development of desensitization of adenylate cyclase by isoproterenol in rat ascites hepatoma AH130 cells. AB - The treatment of AH130 cells with isoproterenol (IPN) resulted in an increase in the activity of cyclic adenosine 3':5'-monophosphate (cyclic AMP) phosphodiesterase with a low Km value and a decrease in IPN-stimulated activity of adenylate cyclase. The activity of cyclic AMP phosphodiesterase was increased by IPN in a dose-dependent manner and reached a maximal increase at 30 min after the addition of 10(-4)M IPN. The addition of mitomycin C (MMC) at 15 min after the initiation of the treatment with IPN inhibited the increase in the activity of cyclic AMP phosphodiesterase in a dose-dependent manner. The extent of the development of desensitization, which was observed as a decrease in IPN stimulated activity of adenylate cyclase, was dependent on the treatment time with IPN and the concentration of IPN. Desensitization was also observed as a decrease in prostaglandin E1-stimulated activity but not in basal and NaF stimulated activity. The combined treatment with IPN and MMC showed a higher IPN stimulated activity of adenylate cyclase than with a single treatment with IPN. This effect resulted from the enhancement of IPN-stimulated activity of the enzyme by MMC itself. These results show that the treatment of AH130 cells with IPN caused an induction of cyclic AMP phosphodiesterase and the development of desensitization of adenylate cyclase. Since the combined treatment with MMC inhibited both phenomena, the high intracellular cyclic AMP level appeared to be maintained by the combined treatment for a longer term than by a single treatment with IPN. PMID- 3025408 TI - Tissue residues, pathology and viral-induced mortality in mice chronically exposed to different cadmium salts. AB - Male Swiss Webster mice were exposed to one of three different cadmium salts (sulfate, acetate and chloride) in the drinking water for 10 weeks and then inoculated with a 14-day LD75 or LD35 dilution of encephalomyocarditis virus. the incidence of viral-induced mortality was lower in 17 of the 18 cadmium-treated groups of mice as compared to controls. Cadmium tissue residues and histopathologic lesions were essentially similar at comparable doses in all groups of mice regardless of the form of cadmium in the diet. PMID- 3025410 TI - Ultrastructure of liver tumors induced in F344 rats by methapyrilene. AB - Liver tumors induced in F344 rats by methapyrilene were studied by electron microscopy. The tumor cells constituting hepatocellular carcinomas showed a pronounced increase in the number of mitochondria and conformational changes of these organelles while the content of lipid, glycogen and smooth endoplasmic reticulum was greatly reduced. The cholangiocarcinomas consisted of bile duct epithelia at varying stages of squamous metaplasia. PMID- 3025409 TI - Studies on nonoxynol-9. IV. Biochemical and morphological effects in the liver, kidneys and lungs of rats following intravaginal and intraperitoneal administration. AB - The content of DNA and collagen was significantly increased in the liver of young adult rats injected intraperitoneally for five consecutive days with Nonoxynol-9 (N-9; 5 mg/100 g). No changes were observed in the lung. A single dose of N-9 increased the activity of SGOT, reaching a maximum 4 to 8 hours after injection. In transmission electron microscopy the hepatocytes showed a dramatic increase of the rough endoplasmic reticulum. Rats injected intravaginally 5 mg/100 g of N-9 daily demonstrated after 15 injections a significant increase of collagen in the liver and kidney. After 15 days, DNA was significantly increased in the kidney. SGOT increased after 5 applications of N-9. Light and electron microscopy of the liver showed signs of nonspecific inflammation. We conclude that N-9 administered to young adult rats intravaginally or intraperitoneally induces biochemical and morphological changes in organs involved in the excretion of this detergent. PMID- 3025411 TI - Enzyme changes in the brain, liver and kidney following repeated administration of mercuric chloride. AB - Alterations in the levels of selected enzymes have been studied in the liver, kidney and brain of mouse following mercuric chloride (1 mg/Kg body wt./d) administration for 10, 20 and 30 d. The activity of acid phosphatase increased in all the tissues, the highest increase was recorded in the kidneys which showed as much as 4.5 fold elevation following mercuric chloride administration for 30 d. Although the alkaline phosphatase activity in the liver and the brain increased following HgCl2 administration, the kidneys experienced a tremendous decline in this enzyme following the same treatment. Mercury-induced changes in ATPase were complex inasmuch as the nature and magnitude of these changes varied with the tissue as well as the duration of the treatment. Whereas the liver ATPase declined after all the treatment intervals, this enzyme increased in the kidney and brain following administration of HgCl2 for 10 d. However, both the kidneys and brain registered a substantial fall in ATPase activity when HgCl2 administration was continued for 30 d. The levels of both glucose-6-phosphatase and succinic dehydrogenase decreased in all the tissues following HgCl2 administration. Invariably, the magnitude of decrease was the highest after 30 d treatment with HgCl2. PMID- 3025412 TI - Regulation of lung fibroblast proliferation and collagen synthesis by alveolar macrophages in experimental silicosis. I: Effect of macrophage conditioned medium from silica instilled rats. AB - Alveolar macrophages were lavaged from silica or saline instilled rats 0, 3, 7 and 14 days after exposure. Macrophages were cultured for twenty-four hours and the conditioned media assayed for the ability to stimulate rat lung fibroblast proliferation and collagen synthesis. Macrophages remained viable throughout the culture period. DNA synthesis was significantly elevated by macrophage conditioned media (MCM) from silica instilled rats (S-MCM) compared to untreated fibroblasts or fibroblasts exposed to MCM from saline instilled control animals (C-MCM). Stimulation of DNA synthesis was not observed when S-MCM was exposed to non-proliferating fibroblasts. Collagen synthesis quantitated as 3H hydroxyproline accumulation or percent collagen synthesis was also increased by day 3 and day 7 S-MCM. Specific activity measurements of intracellular 3H-proline minimized the possibility that the increase in 3H-proline incorporation into collagen was a reflection of increased proline transport. Non-collagen protein synthesis was also increased in fibroblasts exposed to day 14 S-MCM. These results suggest that alveolar macrophages elaborate factors following silica exposure capable of altering the DNA and protein synthetic activity of lung fibroblasts. These changes in fibroblast DNA and protein metabolism are similar to those observed for lung tissue in vivo and lend further support to the hypothesis of macrophage mediation of the pulmonary response following silica exposure. PMID- 3025413 TI - Alterations in phospholipid biosynthesis by polychlorinated biphenyls and polychlorinated biphenylols. AB - Inhibition and stimulation of synthesis of phospholipids by polychlorinated biphenyls (PCB) have been reported by several authors. The mode of action of PCB on the synthesis of phospholipids has not been determined. Results of experiments in the present report indicate that both PCB and polychlorinated biphenylols alter the activities of key enzymes of glyceride and phospholipid synthesis. 4 Chloro-4-biphenylol (CB-OH), 2',3',4',5,5'-pentachlorobiphenylol (PCB-OH) and 2,4,5,2',4',5'-hexachlorobiphenyl (HCB) inhibited sn-glycerol-3-phosphate acyltransferase activity. Inhibition by PCB-OH was noncompetitive with apparent Ki of 2.6 X 10(-4) M. PCB-OH stimulated phosphatidase activity, but no apparent change was observed with CB-OH. PCB inhibited phosphorylcholine glyceride transferase activity, but had no significant effect on diglyceride acyl transferase. PMID- 3025415 TI - Nifedipine attenuates both alpha-1 and alpha-2 adrenoceptor-mediated pressor and vasoconstrictor responses in conscious dogs and primates. AB - Effects of the calcium channel blocker, nifedipine, were examined on the pressor and vasoconstrictor responses to stimulation of alpha-1 and alpha-2-adrenoceptors in chronically instrumented conscious dogs and Rhesus monkeys. Norepinephrine (NE), a mixed alpha-1 and alpha-2 adrenoceptor agonist, phenylephrine (PE) and methoxamine (M), selective alpha-1 adrenoceptor agonists, and B-HT 920, a selective alpha-2 adrenoceptor agonist, were injected i.v. after ganglionic (hexamethonium), beta adrenoceptor (propranolol) and muscarinic receptor (atropine methyl bromide) blockade. In the dog, NE (0.1 microgram/kg i.v.), PE (1 microgram/kg i.v.), M (20 micrograms/kg i.v.) and B-HT 920 (1 microgram/kg i.v.) produced similar increases in mean arterial pressure (NE, 52 +/- 4 mmHg; PE, 42 +/- 4 mm Hg; M, 43 +/- 7 mm Hg; B-HT 920, 45 +/- 6 mm Hg) and total peripheral resistance (NE, 20.8 +/- 5.8 mm Hg/l/min; PE, 23.1 +/- 4.2 mm Hg/l/min; M, 18.2 +/- 2.1 mm Hg/l/min; B-HT 920, 24.8 +/- 7.1 mm Hg/l/min). Nifedipine (0.5 microgram/kg/min i.v.) caused a similar attenuation of the pressor (NE, -54 +/- 8%; PE, -43 +/- 8%; M, -49 +/- 6%; B-HT 920, -56 +/- 8%) and vasoconstrictor (NE, -66 +/- 11%; PE, -52 +/- 9%; M, -60 +/- 13%; B-HT 920, -57 +/- 10%) responses to each of the alpha-adrenoceptor agonists. Nifedipine also attenuated pressor responses to alpha-1 and alpha-2 adrenoceptor agonists similarly in conscious monkeys. Thus, in conscious dogs and monkeys, calcium channel blockade attenuates similarly both alpha-1 and alpha-2 adrenoceptor-mediated vasoconstriction. PMID- 3025414 TI - Activation of excitatory amino acid receptors may mediate the folate-induced stimulation of locomotor activity after bilateral injection into the rat nucleus accumbens. AB - Folic acid (FA) and 5-formyltetrahydrofolic acid (FTHF) have been shown previously to produce a marked stimulation of locomotor activity after bilateral injection into the rat nucleus accumbens. This study was designed to determine whether the hypermotility response produced by the folates is mediated through the activation of excitatory amino acid receptors in the nucleus accumbens. Although FA stimulated locomotor activity, pteroic acid, a congener of FA that lacks the glutamate moiety, was ineffective, suggesting that the glutamate portion of the molecule is essential for the hypermotility response. The N-methyl D-aspartic acid (NMDA) receptor antagonists, D-alpha-aminoadipic acid, DL-alpha epsilon-diaminopimelic acid and MgCl2, at doses that attenuated NMDA-induced hypermotility, were ineffective in decreasing the folate-induced hypermotility response. This behavioral observation is consistent with the biochemical observation that the folates, at a 1 mM concentration, were unable to stimulate the release of [3H]acetylcholine from striatal slices, a model system that is sensitive to the activation of NMDA receptors. In contrast to the ineffectiveness of the NMDA antagonists in inhibiting the response to the folates, the antagonist, glutamic acid diethylester, which inhibited the response to quisqualic acid, but not NMDA, also inhibited the response to both FA and FTHF. Two recently characterized dipeptides, gamma-D-glutamylaminomethylsulfonic acid and gamma-D-glutamyltaurine, antagonized the stimulation of locomotor activity produced by quisqualic acid, FA and FTHF. However, these dipeptides also inhibited the response to NMDA, suggesting that these compounds are not able to distinguish between quisqualate and NMDA receptors in the nucleus accumbens.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3025416 TI - Effects of papaverine and theophylline on renal adenosine transport. AB - We have studied the effects of papaverine and theophylline on the transport of adenosine by the isolated perfused rat kidney and by isolated renal luminal (L) and antiluminal (AL) membrane vesicles. In filtering isolated kidneys perfused with 0.1 mM adenosine, 1 mM papaverine inhibited adenosine removal from the perfusate by 40% and elevated the fractional excretion of adenosine from 0.4 +/- 0.1 to 0.7 +/- 0.0. Theophylline (10 mM) inhibited 0.1 mM adenosine removal in the filtering kidney by 18% and decreased the fractional excretion of adenosine to 0.2 +/- 0.1. In nonfiltering isolated kidneys perfused with 0.1 mM adenosine, 1 mM papaverine or 10 mM theophylline inhibited the rate of adenosine removal from the perfusate by 41 or 19%, respectively. Papaverine (50 or 100 microM) was a potent cis inhibitor of the initial (15 sec) rate of 1 microM [3H]adenosine influx into L and AL membrane vesicles. The influx of 1 microM [3H]adenosine was inhibited partially by 50 microM and inhibited completely by 100 microM papaverine. Papaverine at 10 microM trans stimulated the efflux of 50 microM [3H]adenosine preloaded into AL vesicles. However, theophylline (up to 100 microM) had no effect on the influx of 1 microM [3H]adenosine into L and AL vesicles. The data suggest that papaverine is a potent, direct inhibitor of adenosine transport across both L and AL membranes whereas it is likely that theophylline increases net adenosine influx across the L membrane and decreases adenosine influx across the AL membrane by indirect metabolic effects. PMID- 3025417 TI - Effects of naftidrofuryl on adrenergic nerves, endothelium and smooth muscle in isolated canine blood vessels. AB - Experiments were designed to determine the effects of the vasoactive drug naftidrofuryl on vascular smooth muscle, endothelial cells and adrenergic nerves in isolated canine blood vessels. Naftidrofuryl inhibited contractions of basilar arteries (in a decreasing order of potency), evoked by 5-hydroxytryptamine greater than KCl = anoxia (in rings with endothelium) greater than prostaglandin F2 alpha = uridine-5'-triphosphate. Naftidrofuryl antagonized competitively the contractions evoked by 5-hydroxytryptamine in the femoral artery and the saphenous vein. Naftidrofuryl caused the release of an endothelium-derived relaxing factor(s) from the endothelium of femoral arteries. The compound depressed contractions of saphenous veins evoked by electrical stimulation of the adrenergic nerve endings, but not those caused by the indirect sympathomimetic amine tyramine or exogenous norepinephrine. In saphenous veins incubated previously with [3H]norepinephrine, the drug inhibited the contractions and the release of transmitter evoked by electrical stimulation. Thus, naftidrofuryl acts at different levels in the blood vessel wall to cause: release of endothelium derived relaxing factor(s); inhibition of S2-serotonergic receptors on vascular smooth muscle; prejunctional inhibition of adrenergic neurotransmission; and nonselective inhibition of the contractile process in vascular smooth muscle, which is particularly pronounced in cerebral arteries. PMID- 3025418 TI - Cardiac and vascular actions of decapeptide angiotensin analogs. AB - We have reported previously the characterization of the angiotensin (A) II myocardial receptor and provided biological evidence that the inotropic activities of the octapeptide AII are receptor mediated. In addition to the inotropic activities that this compound demonstrates, it also has potent vascular contractile activities. The decapeptide AI exhibits both cardiac (+)-inotropic and vascular contractile activities. The responses to AI are in large part secondary to conversion to smaller peptides, principally the octapeptide AII. To attempt to separate the inotropic and vascular response to these peptides, several decapeptide A analogs with amino acid substitutions in either the 1, 5 or 7 positions were studied. The compounds were as follows: [Sar1Ile5Ala7]AI; [Sar1Ile5 alpha-MeAla7]AI; [Sar1Val5N-MeAla7]AI and [Sar1Val5Sar7]AI. These analogs were studied in rabbit point-stimulated left atria, isometrically contracting aortic rings and competition for [125I]AII binding to myocardial ventricular membranes. All the peptides exhibited partial AII agonist activities in cardiac and vascular tissues with potencies equivalent to or less than AI. The inotropic and vascular contractile response to the decapeptides was decreased in the presence of the A converting enzyme inhibitor enalaprilat. The inotropic and vascular activities of these compounds in the presence of A converting enzyme inhibitor suggest that A converting enzyme may be responsible for the conversion to smaller peptides. Biologically active compounds were obtained with amino acid substitutions in the no. 1, 5 and 7 positions. Sarcosine substitution in position 1 did not enhance vascular potency as was observed with AII analogs. PMID- 3025419 TI - Decline in beta adrenergic receptor-mediated vascular relaxation with aging in man. AB - Beta adrenergic relaxation of vascular smooth muscle, mediated by cyclic AMP, is blunted with age in a variety of experimental animals. The applicability of these observations to man is uncertain. The dorsal hand vein technique provides an excellent method to examine the direct effects of aging on vascular responsiveness. Thirty-nine healthy male volunteers over the age range of 19 to 79 were studied. No differences in vascular responsiveness to phenylephrine, an alpha adrenergic agonist, were found for either the ED50 (dose producing 50% vasoconstriction) or Emax (maximum vasoconstriction attained). In marked contrast, vascular relaxation induced by isoproterenol, a beta adrenergic agonist, was significantly different in both the ED50 (dose producing 50% of maximum relaxation from a preconstricted state) and Emax (maximum relaxation attained). ED50 +/- S.E.M. for the youngest and oldest deciles were 8.9 +/- 2.3 and 60 +/- 17.0 ng/min, respectively (P less than .05); Emax +/- S.E.M. were 96.7 +/- 3.3 and 37.7 +/- 8.7%, respectively (P less than .001). Nitroglycerin, a smooth muscle relaxant whose effects are not mediated through the cyclic AMP system, was also used to examine the specificity of this blunted response to isoproterenol. Almost complete relaxation was achieved with the infusion of nitroglycerin in the older group. These results suggest that aging is associated with a specific decrease in beta adrenoreceptor-mediated vascular relaxation. PMID- 3025420 TI - Effects of mu-opioid receptor stimulation in the hypothalamic paraventricular nucleus on basal and stress-induced catecholamine secretion and cardiovascular responses. AB - Previous work from this laboratory has demonstrated that opioid peptides, acting at mu-receptors in the brain, stimulate central sympathetic outflow thereby increasing plasma catecholamine concentrations in unstressed rats. Brain sites involved in opioid-mediated catecholamine secretion have not been characterized fully. Additionally, brain opioid effects on sympathoadrenal catecholamine secretion during stress have not been defined. Because the paraventricular hypothalamic nucleus (PVN) plays a central role in autonomic and cardiovascular regulation, we administered the mu-selective enkephalin analog, D-Ala2-NMe-Phe4 Gly(ol)5enkephalin (DAGO), directly into PVN in conscious, unstressed rats and determined the changes in plasma catecholamine concentrations, blood pressure and heart rate. Then, during the peak response, rats were subjected to restraint stress and the same parameters were again measured. Under basal conditions, picomolar doses of DAGO injected into PVN increased plasma concentrations of catecholamines, especially epinephrine, and raised blood pressure. These effects were dose-related (0.01-0.3 nmol) and antagonized by naloxone given either systemically or directly into PVN. Tachycardia was also observed except at the highest dose of DAGO (0.3 nmol). Thus, mu-receptor stimulation in PVN increases central sympathetic outflow in nonstressful situations, producing increased adreno-medullary catecholamine secretion, blood pressure and heart rate. During restraint stress, PVN microinjections of DAGO blunted stress-induced tachycardia, apparently by a vagal mechanism as this effect was blocked by atropine methyl nitrate. PVN DAGO had no significant effect on the plasma catecholamine responses to restraint stress. In contrast, naloxone injected into PVN augmented stress induced epinephrine secretion. Thus, PVN mu-receptors may regulate heart rate during stress, and an endogenous opioid released during restraint stress may modulate adrenomedullary responses to stress. PMID- 3025421 TI - Coronary and myocardial effects of activated neutrophils in perfused rabbit hearts. AB - Studies were conducted in Langendorff-perfused rabbit hearts to assess the actions of phorbol myristate acetate (PMA)-stimulated neutrophils on coronary vascular resistance and left ventricular contraction. Whereas PMA or neutrophils alone were without effects in this preparation, PMA-stimulated neutrophils evoked pronounced increases in coronary resistance and decreases in left-ventricular pulse pressure. The changes in coronary resistance and left ventricular pulse pressure induced by PMA-stimulated neutrophils were attenuated by scavengers of superoxide anion (superoxide dismutase), hydrogen peroxide (catalase) and hydroxyl radical (dimethylthiourea), thereby implying a pivotal role for hydroxyl radical. Inasmuch as the increase in coronary resistance could be reversed partially by the nonspecific vasodilator, sodium nitroprusside, and inasmuch as none of the free radical scavengers influenced PMA-induced neutrophil aggregation, the increase in coronary resistance evoked by PMA-stimulated neutrophils is related in part to oxygen radical-mediated coronary vasoconstriction. PMID- 3025422 TI - Endothelial modulation of vascular relaxation to nitrovasodilators in aging and hypertension. AB - Blood vessel responses to relaxant drugs have been reported to change with aging and with the development of hypertension. In view of the requirement of endothelial cells for the activity of many relaxant drugs, we examined the role of the endothelium in the relaxation response of vascular tissue. Aortic and mesenteric ring segments from normotensive and hypertensive rats, ages 5 to 6, 15 to 18 and 30 to 31 weeks, were examined for relaxation to sodium nitroprusside, sodium nitrite, atrial natriuretic factor and 8-bromo-cyclic GMP. Relaxation responses to the nitrovasodilators were reduced progressively with aging in ring segments of Wistar-Kyoto rats (WKYs) and spontaneously hypertensive rats (SHRs) with intact endothelium; however, intact SHR ring preparations displayed less relaxation to nitrovasodilators at 15 to 18 and 30 to 31 weeks than those of WKYs. Rubbed (endothelium denuded) ring preparations displayed greatest relaxation to nitrovasodilators with no difference being observed between SHR and WKY preparations at any age tested. Relaxation to atrial natriuretic factor and 8 bromo-cyclic GMP was not different between rubbed and unrubbed ring segments or between SHRs and WKYs, indicating no detectable impairment of the overall relaxation response in the vascular smooth muscle of SHRs. These results suggest that the total functional capacity of vascular smooth muscle to relax to nitrovasodilators is not changed with aging or hypertension. However, the endothelial cells exert modulatory influences upon the vascular smooth muscle to reduce overall responsiveness to nitrovasodilators, an effect that is enhanced with aging and the development of genetic hypertension. PMID- 3025423 TI - delta 9-Tetrahydrocannabinol differentially affects sensory-evoked potentials in the rat dentate gyrus. AB - The effects of low doses of delta 9-tetrahydrocannabinol (THC) on auditory-evoked potentials recorded from the outer molecular layer of the dentate gyrus of the rat hippocampus were assessed during performance of an auditory two-tone discrimination task. Conditioned behavior was disrupted up to 2 hr after i.p. injections of delta 9-THC at 1.0 to 2.0 mg/kg: responses to the rewarded stimulus decreased significantly and latency to respond increased significantly. Concurrent recordings indicated that these same doses of THC did not grossly distort the outer molecular layer averaged evoked potential waveform, but did produce opposing alterations in the amplitudes of the previously studied averaged evoked potential components N1 and N2. The amplitude of the N1 component was decreased significantly 0 to 2 hr after THC injection, whereas the amplitude of the N2 component increased significantly. THC injection at 0.5 mg/kg had no significant effects on performance or on N1 or N2 amplitudes. Recovery from the effects of 1.0 to 2.0 mg/kg of delta 9-THC was essentially complete 2 to 4 hr after injection for both measures of performance and for both N1 and N2 amplitudes. The results demonstrate that low doses of delta 9-THC distort behaviorally relevant sensory information converging on the dentate gyrus and support the hypothesis that the psychoactive effects of marijuana may be mediated by action within the hippocampus. PMID- 3025424 TI - Effects of delta 9-tetrahydrocannabinol on sensory-evoked discharges of granule cells in the dentate gyrus of behaving rats. AB - Extracellular action potentials were recorded from identified cells in the dentate gyrus of the awake freely moving rat during performance of a two-tone discrimination task. The effects of low doses of delta 9-tetrahydrocannabinol (delta 9-THC) were assessed on the firing patterns of granule cells to the tone stimuli. Intraperitoneal injections of delta 9-THC at 1.0 and 2.0 mg/kg, but not 0.5 mg/kg, produced a significant suppression of granule cell activity lasting up to 4 hr. This suppression was present in both the spontaneous (pretone) activity and tone-evoked responses of granule cells. The tone responses of cells recorded from the inferior colliculus were unaffected by THC injection at 1.0 and 2.0 mg/kg, implicating further the hippocampus as a site of specific action of delta 9-THC. In the preceding paper it was demonstrated that such doses produced both impairment of discrimination behavior and modifications of sensory-evoked potentials recorded from the dentate gyrus. However, the influence of delta 9-THC on cell firing in the dentate gyrus was more severe both in magnitude and duration of suppression than were the effects on behavior and on sensory-evoked potentials. PMID- 3025425 TI - Effects of mono and divalent cations on total and partial reactions catalysed by pig kidney Na,K-ATPase. AB - Interactions of Na, K, Ca and Mg ions with partially purified Na,K-ATPase from pig kidneys were investigated using as a tool simultaneous determinations of adenosine 5'-triphosphate (ATP)-adenosine 5'-diphosphate (ADP) exchange and ATPase activity catalysed by the enzyme. In the presence of 120 mM-NaCl and 0.5 mM-MgCl2, the effects 12 mM-KCl on ATP-ADP exchange depended on the ATP and ADP concentrations: stimulation (about 10-fold) with 3 mM-ATP-0.75 mM-ADP, no effect with 0.04 mM-ATP-0.01 mM-ADP and inhibition when ATP and ADP concentrations were 0.003 mM and 0.001 mM respectively. The apparent affinities (K0.5) for K stimulation of ATP-ADP exchange and Na,K-ATPase activity were indistinguishable from each other both at 20 mM- and 120 mM-Na. All Na and Na+K-dependent exchanges were completely abolished by 10(-4) M-ouabain. In the absence of K, intermediate Na concentrations were inhibitory of ouabain-sensitive ATP-ADP exchange and ATPase activity. This Na inhibition was more pronounced at low than at high ionic strength and was more noticeable in the exchange reaction. K ions antagonized these Na effects; this K-Na antagonism was more effective on ATPase than on the ATP-ADP exchange. Ca ions, in the absence of Mg, could sustain ATP-ADP exchange. Qualitatively the behaviour of the exchange reaction was similar to that observed with Mg. Quantitatively the rates were always lower with Ca. In the presence of Mg, at low ionic strength and at concentrations which stimulated phosphatase activity, Ca ions were not able to counteract Na inhibition of ATPase and ATP-ADP exchange or to stimulate ATPase activity in the presence of Na. This is another indication of the different reactivity of the K sites involved in phosphatase and ATPase activities as well as of those responsible for the release of Na inhibition of ATPase and exchange reaction. In K-free solutions containing 0.5 mM Na and low ionic strength, Mg was a powerful inhibitor of ouabain-sensitive ATPase and ATP-ADP exchange; the curves relating remaining activities to Mg concentrations were identical for both reactions. When Na was increased to 5 mM, ATP-ADP exchange became more sensitive and ATPase more resistant to Mg inhibition. These data are consistent with (i) a Na-Mg antagonism at the phosphorylation step, and (ii) a Na-Mg synergism in blocking the E2PNa-E1PNa transformation.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3025426 TI - Selective block of inward but not outward rectification in rat sensory neurones infected with herpes simplex virus. AB - Neurones from the dorsal root ganglia of neonatal rat pups were grown in tissue culture and voltage clamped using patch electrodes for whole cell recording. The electrodes were filled with a KMeSO4 solution containing 1.1 mM-EGTA. 15-24 h prior to recording cultures were infected with one of three strains of herpes simplex virus type 1 previously characterized on the basis of a cell fusion assay in cultures of the MRC-5 and BHK-21 cell lines. Infection of sensory neurones with either of the two non-syncitial strains of virus caused a nearly complete loss of the hyperpolarization-activated inward rectifying conductance normally present in these neurones, but did not appear to affect depolarization-activated, calcium-dependent outward currents or voltage-dependent delayed outward rectification. The loss of inward rectification appeared to result from inactivation of the underlying conductance mechanism, rather than from a shift in the gating properties of the conductance mechanism to unphysiological values. In contrast to these results inward rectification was present in sensory neurones infected with another, syncitial, strain of herpes simplex virus. However, quantitative analysis suggested a reduced availability compared to that recorded in uninfected neurones. PMID- 3025427 TI - Forskolin and calcium: interactions in the control of renin secretion and perfusate flow in the isolated rat kidney. AB - Forskolin (activator of adenyl cyclase), high concentrations of K+ and high renal perfusion pressure (manoeuvres known to increase Ca2+ permeability), and calmidazolium (the specific blocker of calmodulin) were used to investigate the mechanisms whereby adenosine 3',5'-phosphate (cyclic AMP) and Ca2+ interact to control renin secretion and perfusate flow in the isolated perfused rat kidney. Forskolin stimulated renin secretion and caused vasodilation in a dose-dependent manner in medium containing 5 mM-Ca2+. Reducing the Ca2+ concentration to 1.25 mM did not affect the renin stimulatory response but blunted the vasodilation. High K+ concentration reversed the forskolin-induced renin secretion and vasodilation. Conversely, forskolin reversed the high K+-induced renin inhibition of renin secretion and vasoconstriction. These effects of forskolin and high K+ were absent when Ca2+ was withheld from the perfusion medium. High renal perfusion pressure also reversed the forskolin-induced renin secretion. Calmidazolium prevented the inhibition mediated by high K+ and high perfusion pressure and thereby restored the forskolin-induced stimulation. Calmidazolium also caused a prompt and marked vasoconstriction. The calmidazolium-induced stimulation of renin secretion was Ca2+-dependent since the drug was ineffective in the absence of Ca2+. On the other hand, the prompt and potent vasoconstriction was present even in the Ca2+-free medium. These results support the hypothesis that cyclic AMP stimulates renin secretion by a mechanism which involves a lowering of membrane permeability to Ca2+ in addition to lowering cytosolic Ca2+ concentration. High K+ and high renal perfusion pressure inhibit renin secretion by raising the membrane permeability to Ca2+, thereby raising the intracellular Ca2+ concentration which then inhibits renin secretion by a calmodulin-dependent process. A further general conclusion from these studies is that membrane permeability to Ca2+ and cellular Ca2+ concentration are of central importance in the control of renin secretion and renal blood flow. PMID- 3025428 TI - Experimental human muscle damage: morphological changes in relation to other indices of damage. AB - The effects of eccentric exercise have been examined in human calf and biceps muscles. Release of muscle creatine kinase and uptake of technetium pyrophosphate have been followed for up to 20 days after the exercise and the results are related to the morphological changes seen in needle biopsy samples. The response to exercise was variable, all subjects developing pain and tenderness in the exercised muscles after 1-2 days and this was followed, in most subjects, by a large increase in plasma creatine kinase 4-6 days after the exercise. This was paralleled by an increased uptake of technetium pyrophosphate into the exercised muscle. Biopsies of the affected muscles showed little or no change in the first 7 days after the exercise but later degenerating fibres were seen, as well as infiltration by mononuclear cells and eventually, by 20 days, signs of regeneration. Very extensive changes were seen in the calf muscle of one subject; changes in the biceps were qualitatively similar but not so severe. In the severely affected calf muscle type II fibres were preferentially damaged. Mononuclear cell infiltration both between and within degenerating fibres was maximal well after the time of peak plasma creatine kinase and it is likely that in eccentrically exercised muscle infiltrating mononuclear cells act to scavenge cellular debris rather than to cause damage to the muscle. PMID- 3025429 TI - Mechanism of vasodilation induced by alpha-human atrial natriuretic polypeptide in rabbit and guinea-pig renal arteries. AB - Effects of alpha-human atrial natriuretic polypeptide (alpha-HANP) on electrical and mechanical properties of smooth muscle cells of the guinea-pig and rabbit renal arteries and of the guinea-pig mesenteric artery were investigated. alpha HANP (up to 10 nM) modified neither the membrane potential nor resistance of smooth muscle cells of the guinea-pig and rabbit renal arteries. In the guinea pig mesenteric and renal arteries, alpha-HANP (up to 10 nM) had no effect on the amplitude and facilitation (mesenteric artery) or depression (renal artery) of excitatory junction potentials nor on action potentials. In the guinea-pig renal artery, alpha-HANP (up to 10 nM) had no effect on the depolarization induced by noradrenaline (NA) (up to 10 microM) but markedly inhibited NA-induced contraction. alpha-HANP (10 nM) slightly inhibited the K-induced contraction. In the rabbit renal artery, alpha-HANP (10 nM) inhibited the NA-induced contraction and to a lesser extent the K-induced contraction. In the rabbit renal artery, the effects of alpha-HANP on the release of Ca from the cellular storage by two applications of NA, and its re-storage, were investigated in Ca-free solution containing 2 mM-EGTA. When 5 nM-alpha-HANP was applied before and during the first application of 0.5 microM-NA, the contraction was markedly inhibited but the contraction to a second application of 10 microM-NA was potentiated. If the first dose of NA was 10 microM the effect was very small. Under the same experimental procedures, nitroglycerine (10 microM) showed almost the same effects as alpha-HANP on the NA-induced contractions. When both the first (3 mM) and second (10 mM) contractions were evoked by caffeine in Ca-free solution, alpha-HANP (5 nM) and nitroglycerine (10 microM) inhibited both contractions to the same extent. In the rabbit renal artery, applications of alpha-HANP or nitroglycerine increased the amount of guanosine 3',5'-phosphate (cyclic GMP) in a dose-dependent manner. However, a much higher concentration of nitroglycerine was required (2 X 10(3) times). In the rabbit renal artery, hydrolysis of phosphatidyl inositol 4,5-bisphosphate (PI-P2) activated by 0.5 microM-NA was inhibited by alpha-HANP, in a dose-dependent manner, but activation by 10 microM NA was not inhibited by alpha-HANP (up to 100 nM).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3025430 TI - Venous mechanoreceptor input to neurones in the inferior mesenteric ganglion of the guinea-pig. AB - Intracellular recordings were made in vitro from neurones of guinea-pig inferior mesenteric ganglia (i.m.g.) attached to the mesenteric vasculature of the distal colon region. The inferior mesenteric vein was cannulated for distension. Non evoked continuous excitatory post-synaptic potentials (e.p.s.p.s) and action potentials were recorded from 38% of neurones in the absence of the colon or any imposed perturbation. Continuous e.p.s.p.s were blocked by hexamethonium, tetrodotoxin and by transection of the lumbar colonic nerves. Distension of the inferior mesenteric vein altered the frequency of continuous activity in 23% of cells which exhibited continuous activity. Venous distension was associated with a depolarization in 31% of cells tested. The depolarization averaged 2.8 mV and in 89% of these was associated with an increase in membrane resistance. A further 14% of cells exhibited an increase in membrane resistance in the absence of depolarization. Venous distension increased the amplitude of fast e.p.s.p.s generated by stimulation of the lumbar splanchnic nerve or the intermesenteric nerve in 38% of cells tested. The results of this study demonstrate the existence of a mechanosensory pathway from the venous bed of the distal mesenteric region to the i.m.g. of the guinea-pig and suggest that distension may enhance neural transmission by increasing the excitability of ganglionic cells. PMID- 3025431 TI - The role of anion transport in the passive movement of lead across the human red cell membrane. AB - Passive Pb transport across the red cell membrane has been studied by measuring Pb uptake from Pb-buffered solutions into resealed ghosts containing EGTA. Over 90% of Pb uptake occurs by a pathway which is inhibited by drugs which block anion transport. The order of effectiveness is 4,4'-diisothiocyanostilbene-2,2' disulphonic acid (DIDS) and 4-acetamido-4'-isothiocyanostilbene-2,2'-disulphonic acid (SITS) greater than phloretin greater than furosemide and bumetanide. Ouabain and cytochalasin B are ineffective. This implicates the anion-exchange mechanism in Pb uptake. The rate of Pb uptake by this route is directly proportional to external Pb2+ and HCO3- concentrations, and inversely proportional to the H+ concentration. These findings suggest that Pb transport depends on the formation of PbCO3 in solution. Pb transport depends upon the presence of a second anion. In the presence of HCO3-, the rate is stimulated in the order ClO4- less than NO3- and CH3CO2- less than F- less than Cl- less than Br- less than I-. The temperature dependence of Pb uptake is similar to that of HCO3-(-)Cl- exchange. Changes in membrane potential appear to influence Pb transport. The effects are small and somewhat variable, but in general a negative internal potential accelerates uptake and reduces exit. A positive internal potential reduces uptake and accelerates exit. These results suggest that Pb is transported on the anion exchanger. Exchange of PbCO3 for a monovalent anion best fits the experimental data, although transport of a ternary PbCO3(-)anion- complex is a possibility. PMID- 3025432 TI - Calcium and long-term transmission damage following anoxia in dentate gyrus and CA1 regions of the rat hippocampal slice. AB - The mechanism of long-term anoxic damage in brain tissue is investigated using the rat hippocampal slice as a model system. The effects of short durations of anoxia on subsequent transmission through two neural pathways are studied. 10 min of anoxia irreversibly abolishes transmission between the perforant path and the dentate granule cells while only 7 min of anoxia irreversibly abolishes transmission between the Schaeffer collaterals and the CA1 pyramidal cells. We examine the involvement of Ca2+ in this irreversible transmission damage and, also, the differential sensitivities of the dentate gyrus and CA1 regions. Substitution of a buffer containing 0 Ca2+ and 10 mM-Mg2+ during the anoxic period substantially improves the recovery of synaptic transmission in both regions of the slice. Dentate gyrus transmission recovers completely after 20 min of anoxia and CA1 transmission survives 10 min of anoxia. These results suggest that Ca2+ influx during anoxia may be an important cause of the long-term damage. The uptake of 45Ca2+ into the intracellular space of the slice is increased during anoxia. This effect is approximately twice as large in CA1 as in the dentate gyrus. Thus, in the dentate gyrus the calculated exchangeable pool of Ca2+ is increased 30% by anoxia and in the CA1 it is increased by 70%. Two incubating conditions which decrease the amount of 45Ca2+ uptake during anoxia protect transmission against long-term damage. (a) Pre-incubation of the slices with 25 mM-creatine elevates tissue phosphocreatine and attenuates the fall in adenosine 5'-triphosphate (ATP) during anoxia. This is associated with partial protection against transmission damage and an approximate 50% attenuation of the anoxic uptake of 45Ca2+. (b) Inclusion of 2 mM-cobalt in the buffer reduces the normoxic uptake of 45Ca2+ so that the uptake during anoxia is no greater than normoxic uptake in the absence of cobalt. This is associated with a complete protection against long-term transmission damage following 10 min of anoxia in the dentate gyrus. A kinetic analysis of the 45Ca2+ uptake shows that the anoxic uptake results primarily from inhibition of the unidirectional efflux of Ca2+ from the cells; there is no calculable increase in the undirectional influx. This suggests that anoxia increases Ca2+ uptake by inhibiting one or more Ca2+ extrusion processes and not by opening depolarization-sensitive Ca2+ channels.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3025433 TI - Inhibition of acid formation by epidermal growth factor in the isolated rabbit gastric glands. AB - The effects of epidermal growth factor (EGF) on basal and stimulated (with histamine, dibutyryl cyclic AMP, and high concentrations of K+) acid formation have been studied in isolated glands from the rabbit gastric mucosa. The changes in the accumulation of [14C]aminopyrine [14C]AP have been used as an indirect measurement of acid production in the glands. Unstimulated gastric glands accumulated [14C]AP indicating the existence of basal acid production in these glands, and EGF caused a small but significant reduction in basal [14C]AP uptake. A similar reduction of basal [14C]AP uptake was observed after exposure to omeprazole but not after ranitidine or prostaglandin E2 (PGE2). Histamine, dibutyryl cyclic AMP and K+ caused a strong and dose-dependent stimulation of acid formation by the glands. EGF, like omeprazole, reduced dose-dependently the [14C]AP accumulation stimulated by both histamine and dibutyryl cyclic AMP, while ranitidine and PGE2 reduced histamine- but not dibutyryl-cyclic-AMP-stimulated accumulation of [14C]AP. In the absence of other external stimuli, an increased K+ concentration enhanced [14C]AP accumulation to levels similar to those produced by histamine and this effect was not changed by EGF, ranitidine or PGE2 but was inhibited by omeprazole. We conclude that EGF interferes with the final steps of acid production between cyclic nucleotides and proton pump of the parietal cells. PMID- 3025434 TI - [Ganglionic transmission and peptides]. PMID- 3025436 TI - Successful management of an androgen-induced hepatic adenoma by embolisation of the hepatic artery. PMID- 3025435 TI - Trypanosoma cruzi: structure and transcription of kinetoplast DNA maxicircles of cloned stocks. AB - Restriction endonuclease mapping of the Trypanosoma cruzi kinetoplast DNA maxicircle was performed in nine cloned stocks using maxicircle probes from T. brucei. Analysis of the maxicircle 13-15-kbp encoding region allowed cloned stocks to be divided into three groups: A, B, and C. Parasites from groups A and B had 3% sequence divergence while parasites from group C showed 16-17% sequence divergence with regard to parasites from groups A and B. Cross-hybridization experiments demonstrated that the 23-25-kbp maxicircle divergent region was similar in sequence in group A and B, but different in group C parasites. The high homology of the T. cruzi and T. brucei encoding regions allowed the use of T. brucei probes to detect T. cruzi transcripts, whose overall picture did not differ for parasites from any of the nine assayed clones. PMID- 3025437 TI - A rare, resectable cause of progressive obstructive airways diseases. PMID- 3025438 TI - Combined extradural-intradural metastatic spinal tumour. PMID- 3025439 TI - Paraquat induced radiosensitization of mammalian cells. PMID- 3025440 TI - Block of cleavage of ultraviolet-irradiated DNA with restriction endonucleases. PMID- 3025441 TI - Effects of delta 9-THC, the principal psychoactive component of marijuana, during pregnancy in the rhesus monkey. AB - The effect of delta 9-tetrahydrocannabinol (THC), the principal psychoactive component in marijuana, was studied in pregnant and lactating rhesus monkeys. THC (2.5 mg/kg/d) or vehicle was administered during different periods of gestation, and effects on pregnancy outcome and hormone concentrations during pregnancy were studied. The most obvious effects were observed with administration early in pregnancy; three of five pregnancies aborted within days after the drug injections began, and one pregnancy resulted in a stillbirth at term. The three abortions were associated with a rapid decrease in chorionic gonadotropin and a subsequent fall in progesterone concentrations to nondetectable levels. In the two pregnancies that continued until term, estradiol concentrations were significantly higher than in vehicle control pregnancies. Daily THC administration during the middle or third portion of gestation resulted in lesser pregnancy loss (one premature birth and four live births at term with THC treatment during the middle portion; two premature births and three live births at term with THC treatment during the third portion). All the premature infants died within two weeks of birth. The weights of the infants at birth and weaning were not significantly different for the infants from vehicle control pregnancies and for full-term infants exposed to THC during gestation. Also, no effects on intrauterine growth and development were detected with ultrasound in the drug treated pregnancies. With acute administration, THC readily crossed the placenta at term in rhesus monkeys and was transferred into the milk of nursing mothers. Significant blood levels of THC and depressant effects were observed in both mothers and neonates when the drug was administered to the mothers one hour before birth or during lactation. PMID- 3025442 TI - Bacterial myocarditis secondary to parvovirus enteritis in a puppy. AB - A purulent bacterial myocarditis, secondary to parvovirus enteritis was diagnosed in a 3-month-old St Bernard puppy. The clinical course was of 2 days duration and was characterized by pyrexia, severe vomition, haemorrhagic diarrhoea and dehydration. Post mortem examination revealed a haemorrhagic enteritis and multifocal purulent myocarditis. Histopathological examination proved that the latter was of bacterial origin. It is postulated that this resulted from a bacteraemia secondary to the intestinal lesions caused by parvovirus infection. PMID- 3025443 TI - Failure of chickens to act as sentinels during an epizootic of eastern equine encephalitis in southern New Jersey, USA. PMID- 3025444 TI - Rearing black flies (Diptera: Simuliidae) in the laboratory: mass-scale in vitro membrane feeding and its application to collection of saliva and to parasitological and repellent studies. PMID- 3025445 TI - Studies on the Vibrio cholerae mucinase complex. I. Enzymic activities associated with the complex. AB - Mucinase enzymes were isolated and partially purified from the culture fluid of Vibrio cholerae grown in proteose peptone-colostrum medium. The mucinase complex contained neuraminidase, endo-beta-N-acetylhexosaminidase, nicotinamide-adenine dinucleotidase and proteinases. Traces of phospholipase activity were detected but the complex lacked aldolase activity. PMID- 3025446 TI - Experimental Argentine hemorrhagic fever: myocardial involvement in Cebus monkey. AB - Four Cebus apella monkeys, to test a recently proposed model for testing neurovirulence of Junin virus (JV) strains, were intracerebrally infected with 10(5) LD50 of the XJ clone 3 strain of JV. There were no significant electrocardiographic abnormalities or gross lesions, but all infected monkeys exhibited a varying degree of histologic myocardial lesions including focal lymphoblastic infiltrates, vascular ruptures, and mild interstitial reactive change. One Cebus showed lymphocytic infiltrates in the caudal portion of the A-V node without specific cell involvement. These preliminary results demonstrate cardiac involvement in experimental infection of the Cebus monkey with Junin virus. PMID- 3025448 TI - Regulation of intracellular pH in LLC-PK1 cells by Na+/H+ exchange. AB - Suspensions of LLC-PK1 cells (a continuous epitheliod cell line with renal characteristics) are examined for mechanisms of intracellular pH regulation using the fluorescent probe BCECF. Initial experiments determine suitable calibration procedures for use of the BCECF fluorescent signal. They also determine that the cell suspension contains cells which (after 4 hr in suspension) have Na+ and K+ gradients comparable to those of cells in monolayer culture. The steady-state intracellular pH (7.05 +/- 0.01, n = 5) of cells which have recovered in (pH 7.4) Na+-containing medium is not affected over several minutes by addition of 100 microM amiloride or removal of extracellular Na+ (Na+o less than 1 mM). In contrast, when the cells recover from an acid load (caused by NH4 preincubation and removal), the recovery is largely Na+ dependent and is sensitive to 100 microM amiloride. These results suggest that with resting pH near neutrality, both Na+o/H+i and Na+i/H+o exchange reactions are functionally inactive (compared to cellular buffering capacity). In contrast, Na+o/H+i exchange is activated by an increased cellular acid load. This activation may be observed directly either as a stimulation of net H+ efflux or net Na+ influx with decreasing intracellular pH. The extrapolation of this latter data suggests a "set point" of Na+/H+ exchange of approximately pH 7.0, consistent with the observed resting intracellular pH of approximately 7.05. PMID- 3025447 TI - Nucleoside transport in rat erythrocytes: two components with differences in sensitivity to inhibition by nitrobenzylthioinosine and p-chloromercuriphenyl sulfonate. AB - The sensitivity of nucleoside transport by rat erythrocytes to inhibition by nitrobenzylthioinosine (NBMPR) and the slowly permeating organomercurial, p chloromercuriphenyl sulfonate (pCMBS), was investigated. The dose response curve for the inhibition of uridine transport (100 microM) by NBMPR was biphasic--35% of the transport activity was inhibited with an IC50 value of 0.25 nM, but 65% of the activity remained insensitive to concentrations as high as 1 microM. These two components of uridine transport are defined as NBMPR-sensitive and NBMPR insensitive, respectively. Uridine influx by both components was saturable and conformed to simple Michaelis-Menten kinetics, and was inhibited by other nucleosides. The uridine affinity of the NBMPR-sensitive transport component was threefold higher than for the NBMPR-insensitive transport mechanism (apparent Km for uridine 50 +/- 18 and 163 +/- 28 microM, respectively). The two transport systems also differed in their sensitivity to pCMBS. NBMPR-insensitive uridine transport was inhibited by pCMBS with an IC50 of approximately 25 microM, while 1 mM pCMBS had little effect on NBMPR-sensitive transport by intact cells. pCMBS inhibition was reduced in the presence of uridine and adenosine and reversed by the addition by beta-mercaptoethanol, suggesting that the pCMBS-sensitive thiol group is located on the exterior surface of the erythrocyte membrane within the nucleoside binding site of the transport system. Inhibition of uridine transport by NBMPR was associated with high-affinity [3H]NBMPR binding to the cell membrane (apparent Kd 46 +/- 25 pM). Binding of inhibitor to these sites was competitively blocked by uridine and inhibited by adenosine, thymidine, dipyridamole, dilazep and nitrobenzylthioguanosine. Assuming that each NBMPR-sensitive transport site binds a single molecule of NBMPR, the calculated translocation capacity of each site is 25 +/- 6 molecules/site per sec at 22 degrees C. pCMBS had no effect on [3H]NBMPR binding to intact cells but markedly inhibited binding to disrupted membranes indicating that the NBMPR-sensitive nucleoside transporter probably has a thiol group located on the inner surface of the membrane. Exposure of rat erythrocyte membranes to UV light in the presence of [3H]NBMPR resulted in covalent radiolabeling of a membrane protein(s) (apparent Mr on SDS gel electropherograms of 62,000). Labeling of this protein was abolished in the presence of nitrobenzylthioguanosine. We conclude that nucleoside transport by rat erythrocytes occurs by two facilitated-diffusion systems which differ in their sensitivity to inhibition by both NBMPR and pCMBS. PMID- 3025449 TI - Regions of conservation and divergence in the 3' untranslated sequences of genomic RNA from Ross River virus isolates. AB - The 3' untranslated (UT) sequences of the genomic RNAs of five geographic variants of the alphavirus Ross River virus (RRV) were determined and compared with the 3' UT sequence of RRV T48, the prototype strain. Part of the 3' UT region of Getah virus, a close serological relative of RRV, was also sequenced. The RRV 3' UT region varies markedly in length between variants. Large deletions or insertions, sequence rearrangements and single nucleotide substitutions are observed. A sequence tract of 49 to 58 nucleotides, which is repeated as four blocks in the RRV T48 3' UT region, occurs only once in the 3' UT region of one RRV strain (NB5092), indicating that the existence of repeat sequence blocks is not essential for RRV replication. However, the precise sequence of the 3' proximal copy of the repeat block and its position relative to the poly(A) tail were identical in all RRV isolates examined, suggesting that it has an important role in RRV replication. Nucleotide substitutions between RRV variants are distributed non-randomly along the length of the 3' UT region. The sequence of 120 to 130 nucleotides adjacent to the poly(A) tail is strongly conserved. Getah virus RNA contains three repeat sequence blocks in the 3' UT region. These are similar in sequence to those in RRV RNA but differ in their arrangement. Homology between the RRV and Getah 3' UT sequences is greatest in the 3' proximal repeat sequence block that shows three differences in 49 nucleotides. The 3' proximal repeat in Getah RNA occurs at the same position, relative to the poly(A) tail, as in all RRV variants. The RRV and Getah virus 3' UT sequences show extensive homology in the region between the 3' proximal repeat and the poly(A) tail but, apart from the repeat blocks themselves, they show no significant homology elsewhere. PMID- 3025450 TI - Models for the structure of outer-membrane proteins of Escherichia coli derived from raman spectroscopy and prediction methods. AB - The secondary structure of porin, maltoporin and OmpA protein reconstituted in lipid membranes is determined by Raman spectroscopy. The three proteins have similar structures consisting of 50 to 60% beta-strand, about 20% beta-turn, and less than 15% alpha-helix. Employing a method for structural prediction that accounts for amphipathic beta-strands, folding models are developed for porin and for the segment of OmpA protein incorporated into the membrane. In the model, the OmpA fragment consists of eight amphipathic membrane-spanning beta-strands that form a beta-barrel. Similarly, porin is folded into ten amphipathic membrane spanning beta-strands that are located at the surface of the trimer towards the lipids and eight predominantly hydrophilic strands in the interior. PMID- 3025451 TI - Co-operativity and enzymatic activity in polymer-activated enzymes. A one dimensional piggy-back binding model and its application to the DNA-dependent ATPase of DNA gyrase. AB - The binding of a ligand to a one-dimensional lattice in the presence of a second ("rider") ligand, which binds only to the first ligand (piggy-back binding), is studied. A model derived from this study is used to analyze the effects of co operativity on the reaction rates of enzymes activated by polymeric cofactors that provide multiple binding sites for the enzyme. It is found that in the presence of strong co-operativity, the steady-state reaction rates of polymer activated enzymes can be very different from the Michaelis-Menten paradigm. By adjusting the co-operativity parameters and the binding constants of the ligands, the model can generate apparent auto-catalytic enhancement by substrates at low substrate concentrations and apparent substrate inhibition at high substrate concentrations. The model is shown to be able to explain the differences in the rates of ATP hydrolysis by DNA gyrase in the presence of long versus short DNA molecules and in the presence of long DNA molecules at different gyrase to DNA ratios. PMID- 3025452 TI - Bacillus thuringiensis var. israelensis delta-endotoxin. Nucleotide sequence and characterization of the transcripts in Bacillus thuringiensis and Escherichia coli. AB - The nucleotide sequence of a 1408 base-pair DNA fragment encoding the insecticidal 27,340 Mr delta-endotoxin of Bacillus thuringiensis var. israelensis has been determined by analysis of a recombinant plasmid from Escherichia coli. The hydropathy plot of the protein shows it to be highly hydrophobic, consistent with a postulated cytolytic mechanism of action for the toxin. In addition, the delta-endotoxin transcriptional start points that are used in B. thuringiensis and an E. coli recombinant have been determined. In B. thuringiensis var. israelensis, transcription initiates from a single start point, and gene-specific transcripts are not observed before stage II of sporulation. This is the stage at which delta-endotoxin antigen is first detected, indicating that control of expression is primarily at the transcriptional level for this protein. Analysis of gene-specific transcription in E. coli indicates that at least three start points are utilized in this organism. Interestingly, the highest level of delta endotoxin mRNA is seen during mid-exponential growth of E. coli and the level appears to decrease as the cells enter the stationary phase of growth. PMID- 3025453 TI - Bacillus thuringiensis var. israelensis delta-endotoxin. Cloning and expression of the toxin in sporogenic and asporogenic strains of Bacillus subtilis. AB - A plasmid-borne gene from Bacillus thuringiensis var. israelensis encoding a 27,340 Mr insecticidal delta-endotoxin has been cloned on a bifunctional multicopy plasmid in a wild-type sporogenic strain and two asporogenic mutants of Bacillus subtilis. The delta-endotoxin gene is expressed at a low level during vegetative growth in all three strains, but the synthesis of the toxin increases markedly during the third hour of stationary phase for both the sporogenic strain and an asporogenic mutant containing the OJ lesion. However, in a stage OA mutant, this increase in delta-endotoxin synthesis is not observed. In both the wild-type sporogenic B. subtilis and the asporogenic OJ strain, phase-bright inclusions, resembling the israelensis crystal in appearance, are visible during late stationary phase. The insoluble inclusions from the B. subtilis transformants, consisting solely of the 27,340 Mr polypeptide, were purified by density gradient centrifugation and found to be extremely toxic to Aedes aegypti larvae. After solubilization in alkaline buffer, this polypeptide was also shown to be haemolytic for human erythrocytes and to lyse Aedes albopictus cells with the same LC50 value as native israelensis delta-endotoxin crystals. During stationary phase, novel mRNA species appear in both the wild-type strain and the OJ mutant, but not in the OA mutant, and these appear to be the major gene specific transcripts. Transcriptional mapping of delta-endotoxin-specific mRNA has shown that the same region of initiation is used at a relatively low level in all three strains during vegetative growth. PMID- 3025454 TI - Simple model for the effect of Glu165----Asp165 mutation on the rate of catalysis in triose phosphate isomerase. AB - We present an ab-initio self-consistent field calculation with a 4-31G basis set on a simple model for proton abstraction from hydroxyacetone (a model for dihydroxyacetone phosphate; DHAP) by formate, which is a model for Glu165 in triose phosphate isomerase. Earlier, we showed that the electrophilic groups on the enzyme (the NH3+ of Lys13 and the NH of His95) were essential to efficient catalysis by triose phosphate isomerase. These groups stabilized the enediolate formed by proton abstraction from the DHAP model so that proton transfer from this molecule to Glu165 became likely. In this study, we carry this analysis one step further. First, we re-examine the energy profile for proton transfer, using the fact that our earlier calculations showed that the combined effect of His95 and Lys13 on the reactant DHAP and intermediate enediolate was to make them equal in energy. Then, we analyze the likely effect of changing Glu165 to Asp165 and relate this to experiments on the kinetics of enzyme catalysis by the Glu165--- Asp165 mutant. PMID- 3025455 TI - Evidence for replicative transposition of Tn5 and Tn9. AB - Two basic types of models, conservative and replicative, have been proposed to account for the mechanism of transposition in bacteria. A method was developed to test these models by positive selection of various transposon-promoted events as galactose-resistant colonies from plasmid-containing cells. The results show that recA plays an important role in the transposition of Tn5 and Tn9 in Escherichia coli. All Tn5-promoted events (cointegrates, deletions and transpositions) are suppressed in recA-, and restored in recA+. In the case of Tn9, however, only transpositions (but not cointegrates or deletions) are diminished in recA-. Therefore, the recA function is required for cointegrate formation by Tn5, and for cointegrate resolution by Tn9. Both Tn5 and Tn9 cointegrates segregate transpositions (which can be seen as sectors on indicator plates) in recA+ hosts. In recA-, the unresolved Tn9 cointegrates undergo a second round of cointegrate formation to excise plasmids bearing galactose-resistant deletions. In growing cultures, the proportion of cointegrates declines steadily while transpositions increase so that, in late stages, cultures are rich in transpositions and contain few cointegrates. This explains the failure of previous workers to identify cointegrates as essential intermediates in transposition. Hence, with the exception of the recA requirement, the mechanism of transposition of these composite transposons is not very different from simple transposons like Tn3. It is concluded that transposition of Tn5 and Tn9 is normally a replicative process. PMID- 3025456 TI - Enhancement of bacterial gene expression by insertion elements or by mutation in a CAP-cAMP binding site. AB - The regulatory region (bglR) of the cryptic bgl operon was characterized by DNA sequence analysis and transcription mapping. Bgl(-)-specific transcription was found to occur in both the wild-type Bgl- and mutant Bgl+ cells. However, the steady-state level of bgl RNA was much higher in the Bgl+ mutant than in the wild type. Activation of the bgl operon by insertion sequence-mediated bglR mutations or point mutations in bglR is therefore the result of increased transcription. The ethylmethane sulfonate-induced point mutations in bglR are alterations in a single base in the cAMP binding protein (CAP) binding site, leading to a stronger binding of the CAP-cAMP complex. The IS1 and IS5-mediated bglR mutations analyzed show that the insertion sequences can activate the bgl operon by integration 78 to 125 base-pairs upstream from the transcription initiation site. The role of the insertion sequences in activation of the bgl operon is discussed. PMID- 3025457 TI - Simian virus 40 protein VP1 is involved in spacing nucleosomes in minichromosomes. AB - We have investigated the average nucleosome spacing in the chromatin from several simian virus 40 virion assembly mutants temperature-sensitive in the major capsid protein VP1. Viral assembly intermediates that accumulate in cells infected with mutants that block virion assembly at the propagation step (tsB) have an average nucleosome repeat length similar to that of wild-type SV40 chromatin, approximately 198(+/- 4) base-pairs. This repeat length is longer than that of the host (BSC-40) cellular chromatin, which has a value of 187(+/- 4) base-pairs. In contrast, SV40 chromatin from cells infected with virus containing a mutation that blocks virion assembly at the initiation step (tsC) has a significantly shorter average repeat length of 177(+/- 4) base-pairs. At the permissive temperature (33 degrees C), tsC chromatin has a nucleosome spacing periodicity essentially the same as that of wild-type SV40 chromatin. In addition to possessing a chromatin structure with nucleosomes that are, on the average, closer together, tsC chromatin contains a nuclease-hypersensitive or open region in nearly all molecules, but apparently the same number of nucleosomes. These findings suggest that nucleosomes are deposited initially on newly replicated SV40 chromatin in such a way as to leave the DNA region containing the origin of replication and transcription enhancers uncovered. Subsequent interaction with capsid proteins appears to increase the average nucleosome spacing and consequently to cover the open region for encapsidation. PMID- 3025458 TI - Na/H exchange in cultured chick heart cells: secondary stimulation of electrogenic transport during recovery from intracellular acidosis. AB - Intracellular acidosis is capable of stimulating a rapid amiloride-sensitive Na/H exchange mechanism in the cell membrane of cultured chick heart cells. The sequence of changes of intracellular sodium and potassium contents during recovery from an acid load in heart cells was determined by atomic absorption spectrophotometry and correlated with electrophysiological measurements. Induction of an intracellular acid load by removal of NH4Cl from the bathing solution caused a rapid rise in sodium content that was amiloride-sensitive. Following a peak, sodium content declined concomitant with a rise in potassium content; these changes were ouabain-sensitive and corresponded with a ouabain sensitive membrane hyperpolarization beyond the calculated potassium equilibrium potential. These observations indicate that pHi regulation in cardiac muscle, following an intracellular acid load involves extrusion of H+ by electroneutral Na/H exchange with the consequent rise in Nai stimulating the electrogenic Na/K pump to return Nai to control level. In the presence of amiloride (10(-4) M), the hyperpolarization was slower although still present: this suggests the existence of another sodium uptake mechanism which contributes to stimulation of electrogenic transport. PMID- 3025459 TI - Interaction of ischemia and reperfusion with subtoxic concentrations of acetylstrophanthidin in isolated cardiac ventricular tissues: effects on mechanisms of arrhythmia. AB - The aim of this study was to determine if "ischemia" and/or reperfusion potentiate digitalis toxicity through effects on oscillatory afterpotentials. Isolated canine Purkinje tissue-papillary muscle preparations were studied using standard microelectrode techniques. Tissues were superfused for 10 min with an "ischemic" solution that mimicked hypoxia, acidosis, elevated lactate, zero substrate and normo- or hyperkalemia. Reperfusion with "normal" Tyrode's solution was then reinstated for 60 min. Next, subthreshold oscillatory after potentials were induced with acetylstrophanthidin (ACS) and the protocol was repeated with ACS in all solutions. Without ACS, ischemic conditions with 4 mM KCl caused depolarization of Purkinje and muscle tissues. Reperfusion resulted in hyperpolarization of Purkinje tissue followed by mild depolarization, and then recovery. Purkinje tissue exposed to ischemic conditions with hyperkalemia responded similarly, except that hyperpolarization upon reperfusion was absent. In the presence of ACS, ischemic conditions with 4 mM KCl abolished oscillatory afterpotentials and caused marked depolarization of Purkinje tissue. Reperfusion decreased the coupling intervals and increased the amplitude of oscillatory afterpotentials relative to pre-ischemic levels, and frequently elicited arrhythmic activity. Arrhythmias ceased and tissues recovered by 60 min of reperfusion. Ischemic conditions incorporating hyperkalemia also abolished ACS induced oscillatory afterpotentials and delayed their reappearance upon reperfusion. All other reperfusion responses were similar. This study demonstrates that "ischemic" suppresses oscillatory afterpotential-mediated effects of digitalis in canine Purkinje tissue, whereas reperfusion potentiates oscillatory afterpotential-induced arrhythmias. PMID- 3025460 TI - Primary bilateral breast cancer. AB - At the St. Francis Medical Center during the 20-year period 1960-1980, there were 1,148 primary breast cancers with a 7.1 percent incidence of synchronous and metachronous invasive primary breast cancer. Twenty-one cases (1.8 percent of the total series) illustrate noninvasive cancer of the breast. The essential approaches toward the diagnosis of a contralateral cancer were breast self examination, history and physical examination, and x-ray mammography. Biopsy of contralateral discrete breast masses was advocated. If biopsies were positive for malignancy, definitive surgical management with indicated supplemental oncological therapy were recommended. "Blind focal biopsy" of the contralateral breast was discouraged. The five- to 25-year disease-free survival rate for 82 invasive bilateral breast cancers was 51 percent with a follow-up percentage of 98 percent during an interval of five to ten years. The survival statistics in the primary bilateral breast cancers when compared to results in primary unilateral breast cancers were not as grave as anticipated. PMID- 3025462 TI - Sarcomatoid carcinoma of the kidney. AB - Among 315 cases of parenchymal renal carcinoma resected between 1975 and 1985, 19 cases (6 per cent) of sarcomatoid carcinoma were identified. All patients were symptomatic or had a palpable mass in the flank at hospitalization. Tumor stage generally was advanced at operation and metastases were detected in 8 patients (Robson stage IV), all of whom died after an average postoperative survival of 8 months. In 3 patients there was tumor invasion of the renal vein (Robson stage IIIA), and they died after an average postoperative period of 11 months. Of 4 patients with perinephric fat invasion (Robson stage II) 2 died of cancer after an average survival of 15 months and 2 are alive with no evidence of disease for an average of 73 months postoperatively. In these later 2 cases the sarcomatoid areas constituted less than 5 per cent of the entire tumor and the remaining tumor was low grade carcinoma. In 3 patients adequate followup is not available and 1 was lost to followup. This histological variant of parenchymal cell carcinoma is a high grade malignancy with a poor prognosis. Operative treatment appears to be ineffective in modifying the behavior of the tumor. PMID- 3025461 TI - Uptake of GABA and nipecotic acid in astrocytes and neurons in primary cultures: changes in the sodium coupling ratio during differentiation. AB - The coupling ratio between sodium and GABA or the GABA analog, (RS)- nipecotic acid, in neuronal and glial uptake of GABA or nipecotic acid was investigated as a function of the morphological differentiation of these cell types in primary cultures. Both in neurons and astrocytes a high-affinity uptake of GABA and nipecotic acid was observed regardless of the degree of differentiation. The values of Km were essentially identical in undifferentiated and differentiated cells. On the other hand, Vmax was significantly increased both in 10-day-old neurons compared to 1- and 3-day-old neurons and in astrocytes treated with dBcAMP compared to untreated cells, i.e., Vmax increased as a function of differentiation in both cell types. In neurons as well as in astrocytes the morphological differentiation resulted in an alteration in the coupling ratio between sodium and GABA. From calculated Hill coefficients it could be deduced that the coupling ratio between sodium and GABA was changed from 1 to 2 during differentiation. Similar results were obtained for (RS)-nipecotic acid. PMID- 3025463 TI - Immunosuppressive acidic protein in patients with testicular cancer. AB - Serum immunosuppressive acidic protein was compared to lactic dehydrogenase as a marker for testicular cancer in 54 patients with testicular cancer, 62 with benign urological diseases and 203 healthy controls. The mean value of serum immunosuppressive acidic protein in patients with testicular cancer (598 +/- 293 micrograms. per ml.) was statistically higher than that in patients with benign disease (429 +/- 163 micrograms. per ml.) and healthy controls (368 +/- 73 micrograms. per ml.). There were statistically significant differences in serum immunosuppressive acidic protein levels between controls and patients with stage 2 (p less than 0.0001) or stage 3 (p less than 0.001) testicular cancer, and between those with stage 1 and stage 2 (p less than 0.0001) or stage 3 (p less than 0.001) disease, respectively. The usefulness of immunosuppressive acidic protein as a marker for testicular cancer also was compared to that of lactic dehydrogenase. Immunosuppressive acidic protein and lactic dehydrogenase levels were elevated almost equally in patients with stage 2 or 3 disease (range 71 to 92 per cent). However, immunosuppressive acidic protein levels were elevated in only 3 of 25 patients with stage 1 cancer (12 per cent), compared to 11 of 25 (44 per cent) with elevated lactic dehydrogenase levels. Immunosuppressive acidic protein was correlated better with tumor stage. In conclusion, serum immunosuppressive acidic protein determinations may be useful in patients with testicular cancer for staging, monitoring treatment results and predicting recurrence. PMID- 3025464 TI - Alpha 2-adrenergic receptor levels in obstructive and spastic Raynaud's syndrome. AB - The present study examines the hypothesis that alterations in the activity of alpha 2-adrenergic receptors (A2R) may underlie the clinical vasospasm seen in patients with Raynaud's syndrome. Platelets were isolated from 13 normal subjects, from 50 patients with vasospastic Raynaud's syndrome, and from 20 patients with obstructive Raynaud's syndrome and A2R levels measured. Binding capacity as determined in femtomoles per milligram of protein (fmol/mg of protein) and affinity were measured by Scatchard plot analysis. In a separate experiment normal human platelets were incubated with either buffer, normal serum, or serum from patients with spastic Raynaud's syndrome and A2R levels were measured. A2R levels in normal subjects averaged 112 +/- 18 fmol/mg; in the patients with spastic Raynaud's syndrome, 191 +/- 14 fmol/mg, p less than 0.01; and in the patients with obstructive Raynaud's syndrome, 164 +/- 31 fmol/mg, p greater than 0.05 (ns). Of the patients with spastic Raynaud's syndrome, 26% had values that were less than the mean value of the normal subjects (69 +/- 7 fmol/mg, p less than 0.05). The A2R levels decreased after incubation with serum from patients who had spastic Raynaud's syndrome by 17.4 +/- 3.1 fmol/mg (p less than 0.05). These results indicate that most patients with vasospastic Raynaud's syndrome have increased platelet A2R levels, which may constitute a primary pathophysiologic abnormality underlying this condition. The presence of subnormal A2R levels in a portion of the patients and the finding of a decrease in measurable A2R levels after incubation in serum from patients with spastic Raynaud's syndrome suggests the possibility of receptor modulation as a mechanism for increased cellular receptor synthesis. PMID- 3025466 TI - Papillomavirus antigens in anorectal condyloma and carcinoma in homosexual men. AB - Squamous cell carcinoma of the cervix and vulva has been associated with infection by human papillomavirus. Recent epidemiologic studies have observed more anal carcinoma among single than married men. To investigate this association between human papillomavirus and squamous proliferative lesions of the anus, we studied eight homosexual or bisexual men with recent diagnoses of anorectal carcinoma and six older patients of unknown sexual orientation with similar diagnoses 20 to 30 years ago. Using an immunohistochemical stain for papillomavirus, we found evidence of its presence in the carcinomas of five (63%) of the eight homosexual men and in two (33%) of the six older patients. Only one homosexual patient had the acquired immunodeficiency syndrome. In the possibly immunocompromised patient, human papillomavirus antigen is present in anorectal dysplasia and carcinoma. PMID- 3025465 TI - Outcome of pregnancy in survivors of Wilms' tumor. AB - Outcome of pregnancy was reported by 99 patients who were cured of childhood Wilms' tumor at seven pediatric cancer centers during 1931 to 1979. These patients carried or sired 191 singleton pregnancies of at least 20 weeks in duration. Among the 114 pregnancies in women who had received abdominal radiotherapy for Wilms' tumor, an adverse outcome occurred in 34 (30%). There were 17 perinatal deaths (five in premature low-birth-weight infants) and 17 other low-birth-weight infants. Compared with white women in the United States, the irradiated women had an increased perinatal mortality rate (relative risk, 7.9) and an excess of low-birth-weight infants (relative risk, 4.0). In contrast, an adverse outcome was found in two (3%) of the 77 pregnancies in nonirradiated female patients with Wilms' tumor and wives of male patients. The high risk of adverse pregnancy outcome should be considered in the counseling and prenatal care of women who have received abdominal radiotherapy for Wilms' tumor. PMID- 3025467 TI - Neonatal herpes simplex virus infection occurring in second twin of an asymptomatic mother. Failure of a modern protocol. AB - A case of neonatal herpes that occurred after vaginal delivery in the absence of genital lesions is presented. The mother had a history of drug addiction and genital herpes. Asymptomatic shedding of herpes simplex virus type 2 from the cervix was noted in the second trimester of pregnancy. Despite being followed with a protocol of close surveillance and serial culturing of the genital tract, the patient gave birth to twins, one of whom developed herpes simplex virus type 2 in the postpartum period. This report presents discussion of this case and herpes surveillance protocols. PMID- 3025468 TI - [Effect of intrathecal morphine and beta-endorphin on pituitary-adrenal function following electrical tooth pulp stimulation]. PMID- 3025469 TI - [The effect of induced hyperthermia on kidney function in dogs]. PMID- 3025470 TI - [Combined radiotherapy and resection in carcinoma of the lung]. AB - To evaluate the therapeutic effect of surgery combined with radiotherapy in lung cancer, 385 operative cases, and 308 received combined treatment, were analyzed. The 5-year survival rate of T3 cases among pTNM-stage III cases undergoing surgery was 2.4% (1/42), while that of cases receiving combined radiotherapy was 18.9% (14/74), which showed some effectiveness. The 5 year survival rate of N2 (n3), operation only cases and combined treatment cases were 5.3% (2/38), and 8.9% (8/90), respectively. Long-term survival among cases of extensive mediastinal metastasis (# 1-9) confirmed the effect of combined therapy. Combined therapy effect was recognized in squamous cell carcinoma. PMID- 3025471 TI - [HCG-producing intrahepatic cholangiocarcinoma]. AB - The serum level of human chorionic gonadotropin (hCG) in a 60-year-old man with intrahepatic cholangiocarcinoma was found to be abnormal (44.6 mIU/ml) at the diagnosis. The patient underwent right hemihepatectomy, and the high serum hCG returned to normal 28 days postoperatively. Six months after the operation, the cholangiocarcinoma recurred in the remnant liver and the hCG level was again elevated to 268 mIU/ml. The patient died of liver failure eight months after the operation. Immunohistochemical studies of the resected tumor showed hCG-positive cells in the cancer nest. In this case, it is (believed) that the intrahepatic cholangiocarcinoma produced hCG and secreted it to the serum. PMID- 3025472 TI - [Antibodies to adult T-cell leukemia-associated antigen (ATLA) in hematological and collagen diseases--comparison of the gelatin particle agglutination (PA) and indirect immunofluorescence (IF) methods]. PMID- 3025473 TI - [Interstitial pneumonia in leukemia patients after allogeneic bone marrow transplantation]. PMID- 3025474 TI - [Tissue affinity of pathogens: virus latent infections and their recurrence]. PMID- 3025475 TI - [Pathology of herpesvirus infections]. PMID- 3025476 TI - Comparison of survival of patients with lung cancer between elderly (greater than or equal to 70) and younger (70 greater than) age groups. AB - Results of radiotherapy for lung cancer in the elderly were compared with those in younger patients. A total of 129 patients were treated by radiation therapy with or without chemotherapy. Fifty-six patients (43.4%) were 70 years old or more (elderly group) at the time of treatment, and the remaining 73 patients were below age 70 (younger group). Treatment results in the elderly group were nearly the same as those in the younger group in all stages of the disease. Survival time was longer for patients with performance status (PS) of 0 to 2 than for those with PS 3 or 4 regardless of age (p less than 0.05). The survival time of the elderly group was nearly the same as that of the younger group for squamous cell carcinoma, but somewhat shorter for adenocarcinoma although the difference was not statistically significant. There was no difference in survival between the two groups when they were treated by combined radiation therapy and chemotherapy. Our results suggest that elderly patients can be treated as safely as younger patients by radiotherapy alone or combined radiation therapy and chemotherapy. PMID- 3025478 TI - A case report of mixed mesodermal tumor of the uterine cervix (mixed, heterologous and homologous sarcoma of the uterine cervix). AB - A case of mixed sarcoma of the uterine cervix in a 17-year-old girl is reported. The tumor showed polypoid features resembling sarcoma botryoides, and was histologically composed of chondrosarcoma as a heterologous element and stromal sarcoma as a homologous one. Leiomyosarcoma was also found in the tumor, but striated rhabdomyoblasts were not present. No epithelial component was detected. After a total hysterectomy, the patient was given chemotherapy and is in good condition. PMID- 3025477 TI - Factors influencing the postoperative course 113 patients with early gastric cancer. AB - One hundred and thirteen patients with early gastric cancer operated on during the period from 1967 to 1982 were followed up until 1985; 24 of them died. The 5- and 10-year cumulative survival rates of 99 patients, excluding 14 (12.4%) who died of diseases unrelated to gastric cancer, were 97.8% and 89.1%, respectively. Of the 24 deaths, seven were due to recurrence of gastric cancer, one to pulmonary metastasis found preoperatively and 16 to diseases unrelated to gastric cancer. Recurrence took the form hepatic metastasis in four cases, bone metastasis in two and recurrence in the gastric remnant in one. The metastases were distant in the majority of cases of recurrence, and recurrence characteristically occurred late, with six patients dying more than 5 years and one dying 10 years after surgery. The recurrences were mostly found in patients with poorly differentiated adenocarcinoma. On the other hand, the causes of death in 16 patients were diseases unrelated to gastric cancer, i.e., primary cancer of other organs in six, operative complications, heart diseases, senility, and pneumonia in two each, and a traffic accident and apoplexy in one each. Thus, Many of the deaths were due to primary cancer of other organs. Four patients underwent non-curative resection. One had lung metastasis found preoperatively and the remaining three had positive margins. The latter three did not undergo a second operation, but the causes of their deaths were not recurrence of gastric cancer. It is necessary to follow up patients from the standpoint not only of recurrence of gastric cancer, but also of diseases other than gastric cancer and multiple gastric cancer in elderly patients. PMID- 3025479 TI - [An assessment of angiographic findings after liver biopsy]. PMID- 3025480 TI - [Adrenal arterial embolization of two cases with a metastatic adrenal tumor from a hepatocellular carcinoma]. PMID- 3025481 TI - [The effect of sulphasalazine, sulphapyridine, 5-aminosalicylic acid and prednisolone on superoxide production by human polymorphonuclear leukocytes: in vitro studies]. PMID- 3025482 TI - [Trace elements in hepatocellular carcinoma--comparison between hepatoma tissue and non-hepatoma liver tissue]. PMID- 3025483 TI - [Forensic pathological studies on autopsy of a patient who died after ingesting the liquid fertilizer Hyponex]. PMID- 3025484 TI - [Renal tubular enzymes]. PMID- 3025485 TI - Antihypertensive action of a non-sulfhydryl angiotensin converting enzyme inhibitor (CV-3317) in various hypertensive models. AB - The antihypertensive action of N-[N-[(S)-1-ethoxycarbonyl-3-phenyl-propyl]-L alanyl]-N-(indan-2-yl) glycine hydrochloride (CV-3317), a nonsulfhydryl compound characterized as an angiotensin converting enzyme inhibitor in our previous work, was examined in hypertensive animal models. In 2-kidney, 1 clip hypertensive rats and dogs, CV-3317 (3 and/or 10 mg/kg, p.o.) produced a sustained antihypertensive action of about 15 to 25 mmHg. Daily oral administrations of CV-3317 (1 to 10 mg/kg/day) to spontaneously hypertensive rats (SHR) for 5 weeks produced a sustained antihypertensive action of 20 to 40 mmHg. When CV-3317 (3 mg/kg) was combined with hydrochlorothiazide (10 mg/kg), its antihypertensive action was intensified in potency and duration. CV-3317 (30 mg/kg) induced a slight hypotension (5 to 10 mmHg) in normotensive rats, but had no effect on the blood pressure of 1-kidney, 1 clip hypertensive rats and on that of a low renin type of DOCA/salt hypertensive rat. The antihypertensive activity of CV-3317 was more potent than that of captopril. In pithed SHR, the pressor response induced by an electrical stimulation of the preganglionic sympathetic nerve, but not the pressor response to norepinephrine, was attenuated by both agents (0.3 mg/kg, i.v.). Both agents may exert their antihypertensive action not only primarily by inhibiting the renin-angiotensin system, but also by inhibiting norepinephrine release from the sympathetic nerve terminals indirectly by reducing the formation of vascular angiotensin II. PMID- 3025486 TI - Intrarenal localization of receptors for alpha-rat atrial natriuretic polypeptide: an autoradiographic study with [125I]-labeled ligand injected in vivo into the rat aorta. AB - We examined intrarenal localization of receptors for alpha-rat atrial natriuretic polypeptide (alpha-rANP) by injecting [125I]-labeled ligand in vivo into the rat aorta. We found that the receptors for alpha-rANP are distributed also on the vasa recta of the outer and inner medulla in addition to the previously reported sites, i.e., the renal arteries, renal pelvis, glomeruli, and inner medullary tissues including collecting tubules. In the vascular bundle of the outer medulla, the majority of grains was preferentially localized on the arterial vasa recta. The electron microscopic autoradiography of the glomerulus showed that the binding sites were mainly localized on the foot process of the podocyte. Since alpha-rANP injected into the aorta under physiological conditions was bound to the glomerulus and vasa recta in the kidney, the effect of ANP on these binding sites may be important in the mechanism of natriuresis. PMID- 3025487 TI - Inhibition of angiotensin converting enzyme by CV-3317, a non-sulfhydryl compound. AB - N-[N-[(S)-1-Ethoxycarbonyl-3-phenylpropyl]-L-alanyl]-N-(indan-2- yl)glycine hydrochloride (CV-3317) and its de-esterified products, CV-3317-COOH and CV-3317 (5-OH)-COOH, inhibited rabbit lung angiotensin converting enzyme (ACE) with the IC50s of 1.2 X 10(-7), 4.0 X 10(-8) and 4.9 X 10(-8) M, respectively, angiotensin I (A-I)-induced vasoconstriction of the rat aorta (IC50: 2.6 X 10(-7), 2.6 X 10( 8) and 5.4 X 10(-8) M, respectively), and A-I-induced pressor response of the rat kidney (IC50: 3.9 X 10(-7), 3.5 X 10(-8) and 2.8 X 10(-8) M, respectively). In these 3 experiments, both de-esterified products were 4 to 14 times more potent than captopril. In rats, CV-3317 (0.0138 to 138 mumol/kg, p.o.) inhibited plasma and lung ACEs, and the effects at a dose of 0.46 mumol/kg lasted more than 8 hr. CV-3317 inhibited the A-I-induced pressor action in rats (0.138 to 13.8 mumol/kg, p.o. or 0.046 to 0.138 mumol/kg i.v.) and dogs (0.46 to 4.6 mumol/kg, p.o.) in a dose-related manner. CV-3317 was more potent and longer acting than captopril in these in vivo ACE inhibitions. CV-3317 augmented bradykinin-induced hypotension (dogs) and contraction of the ileum (guinea pigs) less potently than captopril. In spontaneously hypertensive rats (SHR), CV-3317 (3 mg/kg, p.o.) markedly inhibited plasma and tissue (aorta, kidney, lung and brain) ACEs; and when administered daily for 2 weeks, it inhibited the plasma, aorta, kidney and lung ACEs; in particular, it markedly inhibited the aortic ACE. Captopril (30 mg/kg, p.o.) markedly inhibited tissue ACEs and slightly plasma ACE, but its inhibitory effects on tissue ACEs, except for the aorta, were unclear by repeated dosings and its effect on plasma ACE was rather enhanced. Thus, the inhibition of vascular ACE may be particularly important for the antihypertensive effect of the ACE inhibitors, including CV-3317, in SHR. PMID- 3025488 TI - Effects of putative calmodulin antagonists on the binding of [3H]opioid ligands to rat brain membranes: possible involvement of the change in the membrane fluidity. AB - The effects of putative calmodulin antagonists on the binding of [3H]opioid ligands to rat brain membranes were examined. Chlorpromazine, trifluoperazine, N (6-aminohexyl)-5-chloro-1-naphthalenesulfonamide (W-7) and N2-dansyl-L-arginine-4 t-butylpiperidine amide (TI 233, No. 233) inhibited the binding of [3H] [D-Ala2, Met5]enkephalinamide ([3H]DALAMID) in a dose-dependent manner, and this inhibition was not necessarily specific for Ca2+. The order of the inhibitory potency on the binding of [3H]DALAMID was chlorpromazine. W-7, TI 233 and trifluoperazine. From the Scatchard plots, the effects of these compounds on the binding of [3H]DALAMID were biphasic. Low concentrations of calmodulin antagonists caused a small but significant increase in the maximal binding (Bmax), but upon a further increase of the concentration, a significant decrease in Bmax was observable; the apparent dissociation constant for the ligand receptor complexes increased with the increase in the concentration of calmodulin antagonists. TI 233 (1-30 X 10(-6) M) inhibited, somewhat in a ligand selective manner, the binding of [3H]opioid ligands that have been thought to be selective for opioid receptor types. Calmodulin antagonists caused to increase in the fluorescence anisotropy obtained from 1,6-diphenyl-1,3,5-hexatriene-labeled membrane fractions from rat brains, indicating that these compounds were effective for the decrease in the membrane fluidity. The present results indicate that putative calmodulin antagonists can affect the binding of the opioid ligands to rat brain membranes, probably through a mechanism other than one involving calmodulin-related processes. PMID- 3025489 TI - Binding sites of calcitonin gene-related peptide (CGRP): abundant occurrence in visceral organs. AB - Calcitonin gene-related peptide (CGRP) is a novel peptide amide of 37 amino acid residues, which was first identified as the product of alternative processing of RNA transcripts of the calcitonin gene in humans and rats. Using 125I-human CGRP (hCGRP) as the binding ligand and hCGRP or salmon calcitonin (sCT) as the specific inhibitor of binding, it was examined how the receptor-like binding sites distribute among rat tissues including the nervous system, which is already known to contain binding sites in discrete regions. Some visceral organs (liver, spleen and lung) and possibly the bone marrow of Wistar male rats (8-10 weeks old) were found to be relatively rich in these binding sites. The following parameters were calculated by Scatchard analysis of binding data for the cerebellum, spleen and liver; KD (nM) and Bmax (fmol/mg protein) were 0.61 and 408, 1.08 and 858, and 0.89 and 356, respectively. In these three tissues, both hCGRP and sCT were able to completely suppress the specific binding; the IC50s (nM) of hCGRP for the cerebellum, spleen and liver were 2.57, 2.29 and 3.02, respectively, and the IC50s (microM) of sCT 2.69, 0.41 and 1.78, respectively. The results obtained herein strongly suggest the physiological function of CGRP in these visceral organs including bone marrow. PMID- 3025490 TI - Comparison of typical and atypical benzodiazepines on the central and peripheral benzodiazepine receptors. AB - The affinities of typical and atypical benzodiazepines (BZs) for 3H-flunitrazepam (FLU) and 3H-Ro 5-4864, "peripheral" BZ, binding sites in the central nervous system (CNS) were compared. Each dissociation constant (Kd) value of the 3H-FLU binding sites in the cerebellum, cortex, hippocampus and spinal cord showed the similar results, although the binding maximum (Bmax) for the cortex and cerebellum yielded the largest value and the smallest Bmax appeared in the spinal cord. In contrast, Bmax for 3H-Ro 5-4864 binding sites were about three-fold higher in the spinal cord than in the cerebellum. Among the drugs tested, brotizolam demonstrated the highest affinity for 3H-FLU binding sites in the four regions. Typical and atypical BZs did not normally show different affinities for the two BZ receptor subtypes, except for CL 218,872 and Ro 5-4864. A series of compounds including Ro 5-4864, PK 11195, diazepam and brotizolam had high affinity for the 3H-Ro 5-4864 binding sites in the cerebellum and spinal cord. Other BZs did not show affinity for "peripheral" BZ binding sites in the CNS. In conclusion, brotizolam (or atypical BZ) had the binding affinity to both "central" and "peripheral" BZ receptors, like diazepam (a typical BZ). PMID- 3025491 TI - The effect of calcium antagonists on the activation of guinea pig neutrophils. AB - The role of calcium ions in the activation of guinea pig neutrophil functions was examined by evaluating the effects of calcium antagonists. The data presented here show that calcium antagonists inhibit the activation of guinea pig neutrophil functions elicited by N-formyl-methionyl-leucyl-phenylalanine (FMLP) such as chemotaxis, superoxide anion generation and granule enzyme release in a concentration-dependent manner. The concentrations of calcium antagonists demonstrating the inhibition of neutrophil functions may be somewhat different for each function and higher than that of smooth muscle cells. Calcium ionophore A23187 (A23187) caused superoxide anion generation and granule enzyme release of the neutrophils. A23187 also potentiated the FMLP-induced superoxide anion generation and granule enzyme release of the neutrophils. On the contrary, A23187 neither elicited neutrophil chemotaxis nor affected FMLP-induced neutrophil chemotaxis. These results indicate the possibility that the inhibitory effect of calcium antagonists on the activation of neutrophil functions is probably not simply mediated by inhibition of calcium uptake but also by inhibition of a calcium-dependent intracellular target. PMID- 3025492 TI - Prompt effect of progesterone on the adrenergic response of smooth muscles. AB - The contractile effects of epinephrine on the uterus and ductus deferens of the rabbit and the ductus deferens of the monkey were inhibited by the preincubation with progesterone (6.4 X 10(-5) M) for 1 or 3 min in Locke-Ringer solution. Epinephrine relaxed the guinea pig uterus and taenia caecum. The relaxant effects were enhanced by preincubation with progesterone. Their effects were in a dose dependent manner. There was no apparent change in the number and affinity of alpha-adrenergic receptors in the uterus of rabbits and the ductus deferens of guinea pigs during the incubation with progesterone. Progesterone has no direct effect on alpha-adrenergic receptors. All smooth muscles yielded reproducible contractile reactions to Ca2+ when maintained in depolarizing Tyrode's solution containing K+ (40 mmol/l). Their concentration-response curves were inhibited by preincubation with progesterone (6.4 X 10(-5) M), and they were shifted to the right in a concentration-dependent manner. Established Ca2+-induced contractions were rapidly relaxed by the addition of progesterone (6.4 X 10(-5) M). It suggests that progesterone directly affects the plasma membrane and inhibits the voltage-dependent Ca2+ channel and then inhibits smooth muscle contraction. PMID- 3025493 TI - Enhancement of [3H]clonidine binding to rat cerebral synaptic membranes by treatment with arachidonic acid, prostaglandin (PG) D2, PGE2 and PGF2 alpha. AB - Pretreatment of cerebral synaptic membranes with arachidonic acid (10 nM-1 microM) at 37 degrees C for 40 min caused a significant increase in [3H]clonidine binding but pretreatment at 4 degrees C did not. Scatchard analysis showed that 100 nM arachidonic acid pretreatment resulted in a significant elevation of the Bmax value of the low affinity site. PGD2, PGE2 and PGF2 alpha (10-100 microM) pretreatment also enhanced binding. These polyunsaturated fatty acids could be involved in regulation of the alpha 2-adrenoceptor binding in rat cerebral cortical membranes. PMID- 3025495 TI - Intrahepatic calculi associated with cholangiocarcinoma. AB - A unique case of hepatolithiasis associated with cholangiocarcinoma is described. The intrahepatic calculi consisted mainly of cholesterol rather than calcium bilirubinate. A bacteriological study of the intrahepatic and gallbladder bile was negative, though bacterial infection of the bile duct has been considered a main factor responsible for formation of intrahepatic calculi. PMID- 3025496 TI - [The effects of low dose DBcAMP on hemodynamics and metabolism in patients following open heart surgery]. PMID- 3025494 TI - Tamoxifen binding sites in human mammary cancers. AB - Tamoxifen binding sites (TBS) were measured using 3H-tamoxifen, the objective being to evaluate the relationships among TBS and hormone receptors and/or clinical and pathological characteristics in malignant tissues from 60 patients with mammary cancer. TBS were detected in most (96.7 per cent) cancers in the breast tissues, and the mean content and affinity were 569 fmol/mg X protein with Kd: 1.98 nM. There was no significant correlation between TBS and the estrogen receptor and/or progesterone receptor, with respect to positivity or content. However, there was a significant correlation between TBS and histological grading, thereby indicating the differentiation and the proliferative activity in this tissue. The content of TBS was significantly higher in the group with a high grade of malignancy. The TBS content significantly increased in parallel with the degree of malignancy, as related to tubule formation, nuclear pleomorphism and mitotic activity. On the other hand, there was no significant correlation between TBS and age, tumor size, lymph node status or clinical stage. These results suggest the possibility that TBS may be associated with differentiation and cell proliferation in breast cancer tissues. PMID- 3025497 TI - [A case report of adenoid cystic carcinoma of the trachea]. PMID- 3025498 TI - [A study of hypercalciuria in calcium-containing urolithiasis]. PMID- 3025499 TI - Enhancement of production of superoxide anion by human polymorphonuclear leukocytes exposed to products of the HT-29 human colonic adenocarcinoma cell line. AB - Generation of superoxide anion (O2-) by human polymorphonuclear leukocytes (PMNs) in response to stimulation by opsonized zymosan was enhanced about 100% by prior exposure of the PMNs to human colonic adenocarcinoma cells (HT-29 cell line) or their conditioned culture medium. In addition, HT-29 cells produced substances that had an appreciable chemokinetic activity on PMNs. These tumor-secreted substances appeared to act directly on the PMNs rather than indirectly by interacting with nonadherent mononuclear cells, e.g., lymphocytes. Such a priming activity to display enhanced production of O2- was also found in conditioned medium from F344 rat FR3T3 embryonic fibroblasts but not in conditioned medium from HT-29 repolarized cells (by culture in galactose-containing medium) or from nontumorous human colonic mucosa explants. Such active substances may be important in the host-tumor relationship and, therefore, in the outcome of tumor growth. PMID- 3025500 TI - Lymphocytes and antibody in retrovirus-induced feline pure red cell aplasia. AB - The possible role of antibody and T-lymphocytes was investigated in the pure red cell aplasia (PRCA) associated with feline leukemia virus, subgroup C (FeLV-C), infection. In previous studies, erythroid colony-forming cells were undetectable in marrow culture of cats with PRCA. Yet erythroid burst-forming cells (BFU-E) remained, suggesting that BFU-E were able to differentiate in vitro but not in vivo. It was inferred that immunologic suppression may contribute to the pathogenesis of feline PRCA, and the interactions of antibody and T-lymphocytes with erythroid and granulocyte-macrophage progenitors were studied. Incubation of normal or PRCA marrow cells with PRCA serum or IgG concentrated from this serum and then complement (C') failed to decrease hematopoietic colony growth when compared to the results obtained with cultures of marrow cells incubated with C' alone. In crossover coculture studies, T-cells from Safari cats with PRCA had no inhibitory effect on colony growth from normal or autologous PRCA marrow cells. For the determination of whether feline PRCAs were associated with a clonal T cell process, lymphocytes were obtained periodically from glucose-6-phosphate dehydrogenase (Glc-6-PD) heterozygous cats following FeLV-C infection and were expanded with a crude preparation of interleukin-2. The ratios of Glc-6-PD enzyme types in these samples did not change as cats developed anemia, suggesting that the inhibition of erythropoiesis was not associated with the clonal expansion of T-cells. These studies, therefore, do not support the premise that feline PRCA results from the interaction of antibody or T-cells with erythroid progenitors. PMID- 3025501 TI - Modulation of murine neuroblastoma in nude mice by opioid antagonists. AB - Naltrexone, an opioid antagonist, had an inhibitory effect on the growth of murine S20Y neuroblastoma in BALB/c nude mice. Daily injections of 0.1 mg naltrexone/kg, which invoked a receptor blockade for 6-8 hours/day, resulted in 31-92% delay in latency time prior to tumor expression and a 27-49% increase in mean survival time; the magnitude of antitumor response was governed by tumor burden. Inoculation of neuroblastoma (10(6)-2.5 X 10(4) cells) resulted in measurable tumors in 10-13 days and mean survival times of 30-34 days. Immunoreactive beta-endorphin was detected in tumor tissue (39.7 pg/mg protein). Receptor binding assays revealed specific saturable binding of ligands related to delta- and kappa-binding sites, but not for the mu-binding site. These results demonstrate that opioid antagonist modulation of neuro-oncogenesis is not dependent on the integrity of T-cell-mediated immunity and suggest the feasibility of utilizing the nude mouse model in exploring the role of endogenous opioids in human cancers. PMID- 3025502 TI - Endogenous retroviral env expression in primary murine leukemias: lack of xenotropic antigens but presence of distinct mink cell focus-forming env subtypes correlating with ecotropic virus inoculated and mouse strain. AB - The expression of endogenous retroviral env products on primary leukemia cells of mice was studied with the use of a panel of monoclonal antibodies that discriminate between the various classes of murine leukemia viruses [MuLVs; ecotropic, xenotropic, and mink cell focus-forming (MCF)], as well as between various subtypes within each class. Most spontaneous AKR or Friend MuLV (F-MuLV)- or Moloney MuLV (M-MuLV)-induced AKR or NFS mouse leukemia cells expressed no xenotropic viral envelope antigens but always expressed MCF proteins. Spontaneous C58 lymphomas, on the other hand, often expressed xenotropic proteins in addition to MCF proteins. The subtype of MCF envelope antigens present on leukemia cells, as well as on isolated MCF viruses, varied in a reproducible manner, depending on the mouse strain inoculated and the ecotropic virus used (F-MuLV or M-MuLV). Specifically, F-MuLV consistently induced certain type(s) of MCF envelope antigens on leukemia cells of NFS mice, whereas M-MuLV induced different ones. Similar antigenic patterns were found on the MCF viruses isolated from these mice. Furthermore, MCF envelope antigens (on viruses or leukemia cells) induced in NFS mice by M-MuLV differed from those induced in AKR mice. This finding demonstrated a mouse strain influence on the endogenous MCF env sequences expressed following infection by a given ecotropic virus. The endogenous MCF env sequences in mice thus appear to be a set of genes highly expressed during leukemogenesis, with particular ones specifically expressed in a given mouse strain infected with a given ecotropic virus. PMID- 3025503 TI - Neoexpression of ABH and Lewis blood group antigens in human hepatocellular carcinomas. AB - Expression of blood group ABH, Lewis, and sialylated-Lea antigens in human hepatocellular carcinomas and the adjacent nontumorous liver tissues was investigated with the use of seven monoclonal antibodies against these carbohydrate determinants. Chromatogram antibody-binding assay and solid-phase enzyme immunoassay of the upper-phase neutral glycolipids revealed the tumor associated expression of blood group A-active glycolipids incompatible with blood type status of the patients, a blood group A-active glycolipid with mobility on thin-layer chromatography between the known 6- and 8-sugar blood group A-active glycolipids in human erythrocytes, blood group H-active glycolipids, and blocked synthesis of Lea-active glycolipids with or without concomitant accumulation of Leb-active glycolipids. Immunohistochemical analysis of the fixed tissues with the use of an avidin-biotin-peroxidase complex method revealed blood group antigens in biliary epithelial cells but not in parenchymal liver cells. However, hepatocellular carcinoma cells in some cases expressed H and Leb antigens. Although only type 1 chain H antigen was detected in biliary epithelial cells, both type 1 and type 2 chain H antigens were found in hepatocellular carcinoma cells. PMID- 3025504 TI - Epidemiology of borderline ovarian tumors. AB - Ovarian tumors of low malignant potential, often termed "borderline tumors," have been defined as those that have some but not all of the morphologic features of malignancy (i.e., they are not invasive). With the use of data obtained by the western Washington population-based Cancer Surveillance System for 1975-83, the incidence of serous and mucinous borderline epithelial ovarian tumors was analyzed, as well as the survival of women who developed them. The incidence of borderline tumors increased with increasing age, although at a pace somewhat slower than that of malignant ovarian tumors. There was an upward trend in the incidence of borderline tumors starting in the late 1970's, a trend not present for malignant tumors. Only 12% of borderline tumors were not confined to the ovary, as opposed to 40% of malignant Grade I and 73% of other malignant ovarian neoplasms. At 5 years following diagnosis, the survival of women with borderline tumors was 93% that of the general female population. This percentage varied little by stage or histologic type. Given the reduced survival of women with these ovarian tumors and the lack of a sharp histologic distinction between borderline and Grade I malignant lesions, it is recommended that borderline ovarian tumors be routinely ascertained by population-based cancer registries. PMID- 3025505 TI - Products of cells cultured from gliomas. V. Cytology and morphometry of two cell types cultured from glioma. AB - Explants of cells of a human glioma were evaluated with the nuclear fluorochrome 4',6-diamidino-2-phenylindole, by phase-contrast illumination, and by Giemsa staining correlated with double immunofluorescence for glial fibrillary acidic protein (GFAP) and fibronectin (FN). FN-positive (FN+) cells lacked GFAP detectable by immunofluorescence. Their mean nuclear-to-cytoplasmic ratio was large (0.192). Actual mean areas of nuclei (1,252 microns2) and cytoplasm (8,376 microns2) of FN+ cells compared with mean areas of fibroblasts suggested that the high nuclear-to-cytoplasmic ratio of FN+ cells was due to their microscopically evident reduced cytoplasmic spreading rather than to larger nuclei. Some FN+ cells showed marked variation in nuclear and nucleolar size and shape. Others had abnormal mitoses or hyperchromatic nuclei. GFAP-positive (GFAP+) cells lacked FN detectable by immunofluorescence. GFAP+ cells were smaller and less round than FN+ cells. Their usual location was growing on a layer of FN+ cells. The mean nuclear-to-cytoplasmic ratio (0.245) of GFAP+ cells was the highest in the study, surpassing the ratio of the continuous glioma line LM (0.176). Mean areas of nuclei (289 microns2) and of cytoplasm (1,350 microns2) of GFAP+ cells suggested that their high nuclear-to-cytoplasmic ratio was due to their microscopically evident reduced cytoplasmic spreading. Reduced spreading was associated with extension of long, thin cytoplasmic processes. The majority of GFAP+ cells showed marked cytoplasmic basophilia, nuclear hyperchromasia, and clumped chromatin. Features observed in both FN+ and GFAP+ cells from this high-grade astrocytoma are features associated with malignant transformation in more thoroughly studied tumor systems. PMID- 3025506 TI - Histopathology of lung cancer in New Mexico, 1970-72 and 1980-81. AB - In conjunction with a population-based case-control study of lung cancer in New Mexico, the histopathology of cases diagnosed during 1980 and 1981 and during 1970-72 was reviewed. Adequate histologic or cytologic material was obtained for 725 cases, with 308 during 1970-72 and 417 during 1980-81. The light microscopic histologic type was classified on the basis of review by 2 pathologists. No significant differences were found in the histologic-type distributions in Hispanics and non-Hispanic whites. In males, the distributions of histologic types were similar in the two time periods, but in non-Hispanic white women the proportion of adenocarcinoma declined during 1980-81 as the proportion of small cell carcinoma increased. The panel classification was compared with that recorded by the New Mexico Tumor Registry. Overall agreement was 52.1% for 1970 72 and increased to 65.2% for 1980-81. The discrepancies between the two classifications were largest for the categories of large cell undifferentiated carcinoma and "other malignancy." PMID- 3025507 TI - [Reconstructive-plastic operation in recurrence of a bronchial adenoma]. PMID- 3025508 TI - [Diagnosis and treatment of bronchial adenoma]. PMID- 3025509 TI - [Studies of the role of human Cytomegalovirus infection in the etiology of retinitis and choroiditis. Preliminary report]. PMID- 3025510 TI - Effect of angiotensin I converting enzyme inhibition on circulating atrial natriuretic peptide in humans. AB - We investigated the effects of inhibition of angiotensin II (ANG II) formation with an ANG I converting enzyme inhibitor, namely Enalapril, on circulating levels of atrial natriuretic peptide (ANP). Seven normal volunteers received a low sodium diet in order to stimulate the renin-angiotensin system (period I). Subsequently, subjects maintained their low dietary sodium intake and received Enalapril perorally (period II). Each study period lasted for 6 days. Results indicated that subjects maintained a low dietary sodium intake with expected changes of the renin system before and during ANG I converting enzyme inhibition. Enalapril elicited a substantial fall in circulating ANP levels in six of the seven subjects studied. The causes of this change will have to be established but may possibly relate to hemodynamic consequences of Enalapril. PMID- 3025511 TI - Atriopeptin III induces endothelium-independent relaxation and increases cGMP levels in rabbit aorta. AB - Atriopeptin III (AP III) is a 24 amino acid synthetic peptide and a fragment of the atrial natriuretic factor. Using isolated rabbit aortic segments with intact or functionally destroyed endothelium, the effect of AP III on muscular relaxation was examined. cAMP- and cGMP levels were also determined. Aortic segments were pre-contracted with norepinephrine 10(-8) M, angiotensin II 10(-7) M or potassium chloride 20 mM. Addition of AP III (10(-10)-10(-7) M) to these pre contracted segments exerted a concentration-dependent relaxation which was independent of intact endothelium, with EC50 values of 2.2 X 10(-9) M, 2.0 X 10( 9) M and 3.1 X 10(-8) M, respectively. In aortic segments with functionally destroyed endothelial surface, basal levels of cGMP were lower compared with intact vascular tissue. The process of relaxation, induced by AP III, was associated with marked increases of cGMP in intact vascular tissue. cAMP levels were unchanged in both preparations. Our results suggest that AP III elicits a direct, endothelium-independent vascular relaxation associated with increased levels of cGMP in tissue with intact endothelium. Extrusion of intracellular Ca++ by activation of cGMP-dependent protein-kinase may be part of the vasorelaxant profile of AP III. PMID- 3025512 TI - [What is meant by angiotensin converting enzyme inhibitors?]. PMID- 3025513 TI - Neutrophil metabolic dysfunction in genetically heterogeneous patients with glycogen storage disease type 1b. PMID- 3025514 TI - Hepatocarcinogenesis: a dynamic cellular perspective. PMID- 3025515 TI - Rocky Mountain spotted fever and sensory neuropathy. PMID- 3025516 TI - Site of gossypol inhibition of steroidogenesis in purified mouse Leydig cells. AB - Gossypol, a phenolic compound that has been studied as a potential male contraceptive, inhibits basal and LH-stimulated testosterone release from Leydig cells in vitro. The present study investigates the mechanism of this inhibition using preparations of purified mouse Leydig cells. Gossypol inhibited LH stimulated progesterone, 17-hydroxyprogesterone and androstenedione production by mouse Leydig cells incubated in vitro. It also inhibited dibutyryl cyclic AMP stimulated production of testosterone but was without effect in the presence of added pregnenolone or 25-hydroxycholesterol. These results suggest gossypol exerts its major inhibitory effect on Leydig cell function at a point between LH dependent stimulation of cyclic AMP dependent protein kinase activity and increased availability of cholesterol for side chain cleavage. PMID- 3025517 TI - ACTH and thyroid hormone regulation of 3 beta-hydroxysteroid dehydrogenase activity in human fetal adrenocortical cells. AB - The regulation of 3 beta-hydroxysteroid dehydrogenase, delta 4,5-isomerase (3 beta-HSD) and hydroxysteroid sulfotransferase (HST) activities by ACTH and thyroid hormones was investigated in cell cultures from the fetal zone or definitive zone of the human fetal adrenal cortex, using a serum-free, defined medium. ACTH alone maximally stimulated the 3 beta-HSD activity several-fold, whereas triiodothyronine (T3) alone had no effect on this enzyme activity in cell cultures from each zone. However, treatment of cultures with maximal concentrations of 10 nM ACTH plus 1 nM T3 significantly increased the 3 beta-HSD activity an additional 59-115% over that for ACTH alone, without alteration of the ACTH ED50 (0.3 nM). The T3 ED50 was 31 pM for this interaction with ACTH. Thyroxine at a reduced sensitivity had the same interaction with ACTH. T3 similarly increased the stimulation of 3 beta-HSD activity by the steroidogenic agents, cholera toxin and a cAMP analog. The HST activity was not affected by T3 alone but was stimulated by ACTH alone. This stimulation was an order of magnitude less than that for the 3 beta-HSD activity in the same cultures. ACTH plus T3 did not have the synergistic effect on HST activity as observed for the 3 beta-HSD activity. These studies show an interaction between ACTH and thyroid hormone for the stimulation of 3 beta-HSD activity in cell cultures of the human fetal adrenal cortex. PMID- 3025518 TI - Biological activity of vitamin D metabolites and analogs: dose-response study of 45Ca transport in an isolated chick duodenum perfusion system. AB - We have previously reported that vascular perfusion of the normal vitamin D3 replete chick duodenum with physiological amounts of 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] increases the unidirectional movement of 45Ca from the lumen to the venous effluent under conditions of normal (0.9 mM) Ca2+ concentrations in both the lumen and vascular perfusate [Endocrinology 115: 1476 1984)]. The purpose of the present study was to determine the dose responsivity of this perfused intestinal calcium transport system for 1,25(OH)2D3 and some structurally related congeners. The dose-response curve was biphasic for all compounds studied; for 1,25(OH)2D3 initial stimulation of transport was detected at only 30 pM [the plasma concentration of 1,25(OH)2D3 is normally 125 pM] while maximal stimulation was 154% above control at a concentration of 650 pM. Above 650 pM 1,25(OH)2D3 the stimulation fell off sharply and transport had returned to basal levels by 1.3 nM. The relative potency of the D homologs tested was respectively 1,25(OH)2D3: 10,000; 1-alpha-hydroxyvitamin D3: 400; 25-hydroxyvitamin D3: 200; 24R,25 dihydroxy-vitamin D3: 137; vitamin D3: 34; 5,6-trans-25-hydroxyvitamin D3: 3. These results establish the usefulness of the perfused intestinal calcium transport system to study the nongenomic actions of 1,25(OH)2D3 on intestinal calcium transport. PMID- 3025519 TI - Inhibition of pituitary-gonadal function in male rats by a potent GnRH antagonist. AB - The inhibitory effects of the potent GnRH antagonist, [Ac-D-pCl-Phe1,2,D-Trp3,D Arg6,DAla10]GnRH (GnRHant) upon pituitary-gonadal function were investigated in normal and castrated male rats. The antagonist was given a single subcutaneous (s.c.) injections of 1-500 micrograms to 40-60 day old rats which were killed from 1 to 7 days later for assay of pituitary GnRH receptors, gonadal receptors for LH, FSH, and PRL, and plasma gonadotropins, PRL, and testosterone (T). In intact rats treated with low doses of the antagonist (1, 5 or 10 micrograms), available pituitary GnRH receptors were reduced to 40, 30 and 15% of the control values, respectively, with no change in serum gonadotropin, PRL, and T levels. Higher antagonist doses (50, 100 or 500 micrograms) caused more marked decreases in free GnRH receptors, to 8, 4 and 1% of the control values, which were accompanied by dose-related reductions in serum LH and T concentrations. After the highest dose of GnRHant (500 micrograms), serum LH and T levels were completely suppressed at 24 h, and serum levels of the GnRH antagonist were detectable for up to 3 days by radioimmunoassay. The 500 micrograms dose of GnRHant also reduced testicular LH and PRL receptors by 30 and 50% respectively, at 24 h; by 72 h, PRL receptors and LH receptors were still slightly below control values. In castrate rats, treatment with GnRHant reduced pituitary GnRH receptors by 90% and suppressed serum LH and FSH to hypophysectomized levels. Such responses in castrate animals were observed following injection of relatively low doses of GnRHant (100 micrograms), after which the antagonist was detectable in serum for up to 24 h. These data suggest that extensive or complete occupancy of the pituitary receptor population by a GnRH antagonist is necessary to reduce plasma gonadotropin and testosterone levels in intact rats. In castrate animals, partial occupancy of the available GnRH receptor sites appears to be sufficient to inhibit the elevated rate of gonadotropin secretion. PMID- 3025520 TI - Estradiol membrane binding sites on human breast cancer cell lines. Use of a fluorescent estradiol conjugate to demonstrate plasma membrane binding systems. AB - A fluorescent estradiol macromolecular complex was used to study and to characterize steroid binding to membranes of living target cells. Ligand binding to plasma membranes was quantitated with a sensitivity of 0.1 nM. In this way, we found two types of estradiol-binding sites on hormone sensitive MCF-7 cells. Type A sites (8000-16000 sites per cell) were rapidly saturated at low concentrations of the estradiol-bovine serum albumin-fluorescein isothiocyanate macromolecular complex (E2-BSA-FITC). They had a greater affinity for the complex than did the type B sites for which a phenomenon of cooperative fixation was shown. The complex binding was displaced by estrogenic molecules, but not by non-estrogenic compounds, such as cortisol or progesterone. We also studied complex binding on another breast cancer cell line, MDA-MB-231 (MDA), without intracellular estrogen receptors. These cells showed a specific plasma membrane binding system for estrogen, but lacked the high affinity type A binding site. Then, we report the effects of enzyme treatments (trypsin, phospholipase A2 and neuraminidase) on E2 BSA-FITC binding to MCF-7 cell membranes. The quantity of complex bound to membranes decreased after phospholipase and neuraminidase treatments and increased after trypsin. But, in the three cases, the binding was no longer specific because it could not be displaced by E2-BSA or by estradiol. The enzymatic effects were reversible and specific binding was totally restored within 24 h. However, in the presence of the protein synthesis inhibitor, cycloheximide, no restoration of specific binding occurred on trypsin-treated cells. Estrogen binding to MCF-7 and MDA cell plasma membranes thus possesses the three characteristics of all mediated transport processes across biological membranes: saturability, substrate specificity, and specific inhibition. However, the high affinity type A binding site was found only on the estrogen-sensitive cell line, MCF-7. PMID- 3025521 TI - Effect of ketoconazole on placental aromatase, 3 beta-hydroxysteroid dehydrogenase-isomerase and 17 beta-hydroxysteroid dehydrogenase. AB - Ketoconazole, an orally-active, broad spectrum mycotic agent, was shown to inhibit in vitro human placental microsomal aromatase but was without effect on 3 beta-hydroxysteroid dehydrogenase-isomerase (3 beta-HSD-I) and 17 beta hydroxysteroid dehydrogenase (17 beta-HSD) activities. The Km of placental aromatase for testosterone was 30 +/- 1.1 nmol/l (mean +/- SEM, n = 6). Inhibition (determined by Lineweaver-Burk plot) was non-competitive with respect to substrate with a Ki value of 3.0 +/- 1.4 mumol/l (mean +/- SEM, n = 6). Ketoconazole was without effect on the 3 beta-HSD-I and 17 beta-HSD activities when using [3H] pregnenolone and [3H] oestradiol, respectively, as substrates. Since ketoconazole is known to inhibit cytochrome P-450-dependent enzyme reactions, the results of the present study support the contention that cytochrome P-450 is involved in the aromatisation process. PMID- 3025522 TI - Lack of effect of acute alcohol ingestion on erythrocyte Na+, K+ -ATPase activity or passive sodium uptake in vivo in man. AB - Erythrocyte ouabain-sensitive 86rubidium uptake (as an index of Na+,K+-ATPase activity) as well as passive sodium uptake were measured in a group of young men in response to drinking either nonalcoholic beer (as a control) or the same drink with alcohol (1 ml/kg) added. Plasma alcohol concentration rose to 16.7 mM within 70 min of commencement of drinking. There was no change in uptake of 86rubidium or sodium after 60 min in either the alcohol or control studies. There was a late increase in plasma sodium levels 90 min after alcohol ingestion, attributed to fluid volume contraction following diuresis. In contrast, plasma potassium levels fell after alcohol ingestion. This was associated with a decrease in urinary potassium excretion, hence ascribed to an intracellular shift of potassium ions, a change inconsistent with NA+,K+-ATPase inhibition. It is concluded that acute moderate doses of alcohol do not influence NA+,K+-ATPase activity or passive sodium uptake in circulating erythrocytes. Unless vascular smooth muscle is more sensitive to the effects of alcohol in vitro, these findings make it less likely that inhibition of NA+,K+-ATPase mediates alcohol-related hypertension in man. PMID- 3025523 TI - [Effect of chronic chloroquine poisoning on the cardiac binding sites of calcium inhibitors in the rat]. AB - Among the antimalarial drugs, chloroquine (CLQ) and 4-aminoquinoline derivatives display important interferences with biological membranes. The present study reports the effects of CLQ on dihydropyridine binding sites, measured in the heart of rats intoxicated with the drug. Acute intoxication does not entail any modification of the sites. On the other hand, binding sites were dramatically decreased by a chronic intoxication realized twice a week. In such a case, this decrease may be directly related to CLQ concentration in the heart. The regulation mechanisms involved are discussed. PMID- 3025524 TI - [Effect of GABA-linoleamide and glycine linoleamide on pentamethylenetetrazole convulsions]. AB - We have studied the effect of GABA-linoleamide (GL) (100 mg/kg i.p.) and glycine linoleamide (LG) (70 mg/kg i.p.), on the pentamethylenetetrazole (PTZ) (82 mg/kg i.p.) convulsions and lethality of rats. LG antagonize more efficiently than GL the PTZ convulsions and lethality. In another experiment, rats have received haloperidol (2 mg/kg i.p.) for 12 days. Four days after the last administration of haloperidol, rats received as previously GL (100 mg/kg i.p.) or LG (70 mg/kg i.p.) and PTZ (82 mg/kg i.p.). GL only antagonized the PTZ convulsions and lethality. The above results seem to demonstrate the importance of GABA-ergic and glycinergic receptors in the control of PTZ convulsions and they are supported by the recent data on the role of the inhibiting neuromediators, GABA and glycine, in the substantia nigra. PMID- 3025525 TI - Cimetidine does not alter free unchanged captopril pharmacokinetics and biological effects in healthy volunteer. AB - The effects of cimetidine (single or repeated administration) on free unchanged captopril plasma levels, pharmacokinetic parameters and plasma converting enzyme inhibitory effects have been investigated in normal healthy volunteers. Cimetidine affected neither captopril pharmacokinetic parameters nor its biological effects, suggesting that no change in captopril dosing is necessary when cimetidine is co-administered. PMID- 3025526 TI - Introns as relict retrotransposons: implications for the evolutionary origin of eukaryotic mRNA splicing mechanisms. AB - A model is presented for the evolutionary origin of intron sequences within eukaryotic protein-coding genes. We propose that introns are the vestiges of transposable elements and, specifically, that they represent a novel class of retrovirus-like transposons. The attraction of the retrotransposon model is that it gives the RNA splicing mechanism a central role in the evolution of introns. There is a growing body of evidence to suggest that several aspects of splicing are intron-encoded. Consequently, it is reasonable to look for evolutionary explanations of the splicing mechanism in the context of the evolution of the intron sequences themselves. According to this model the ancestral intron genomes were replicated into RNA copies simply because of their insertion within transcriptionally active regions of the host genome. Splicing was necessary not only to minimize their negative effects on host gene expression, but also, and perhaps more importantly, to generate new copies of the intron genome free of flanking exon sequences. These spliced intron copies were then available for reverse transcription and reinsertion elsewhere in the genome. Thus, splicing can be seen as an essential step in the intron replication cycle. Most modern introns have probably lost the majority of their original genetic content and may be considered as degenerate evolutionary relicts. An exception to this degeneracy is the set of splicing signals which must be retained because of its continued importance to host cell survival.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3025527 TI - Presence of circulating endotoxins during cardiac operations. AB - Ten patients having coronary artery bypass grafting were intraoperatively and postoperatively analyzed for endotoxins with the Limulus amoebocyte lysate test. A new highly sensitive rocket immunoelectrophoretic assay for reading the reactions of endotoxins with Limulus amoebocyte lysate was used. Preoperatively, all blood samples from the patients had negative Limulus amoebocyte lysate tests, negative blood cultures, normal total white cell counts, and were clinically without signs of infection. Intraoperatively, a substantial amount of endotoxins were found in samples from the extracorporeal circuit, the pulmonary artery, and the cardiac suction lines, which persisted during the cardiopulmonary bypass. The endotoxin content decreased significantly (p less than 0.05) 6 hours after cardiopulmonary bypass and further decreased within the seventh postoperative day (p less than 0.01). A positive Limulus amoebocyte lysate test was also found in some of the fluids administered during the operation, that is, the cardioplegic fluids, the priming fluids for the extracorporeal circuit, the blood transfusions, and the ice for local cooling. Postoperatively, all patients had rectal temperatures below 38.5 degrees C, but no correlation was found between the magnitude of endotoxin content and the degree of fever. Only one of the patients had positive blood cultures. Despite the measured endotoxin content, no intraoperative or postoperative complications were found. PMID- 3025528 TI - Structure-activity relationships of aromatic retinoids on the differentiation of the human histiocytic lymphoma cell line U-937. AB - The differentiation-inducer activity of a series of derivatives modified on the terminal ring, the polyene side-chain or the polar end group of the retinoic acid molecule was tested on the human histiocytic lymphoma cell line U-937 and compared with that of all-trans-retinoic acid. Only retinoids with both an unsaturated terminal end ring and a free carboxyl polar end group were found to be active in this system. Introduction of an aromatic ring in place of the first double bond of the side chain increased highly the activity whereas cyclisation of the last two double bonds decreased it. Replacement of the unsaturated terminal ring by an aromatic ring abolished the activity as did the esterification of the carboxyl end group or its replacement by a sodium sulfinate, sodium sulfonate or ethyl sulfone end group. All but only the active retinoids induced the same morphological and biochemical changes on U-937 cells, suggesting that they have the same route of action. However, biologically inactive retinoids were shown to be able to inhibit the differentiation of U-937 cells, induced by active ones, indicating that they can compete for a common "receptor". PMID- 3025529 TI - Dexamethasone caused alterations in poly(A) polymerase levels of a human leukemic cell line. AB - The effect of glucocorticosteroids upon the poly(A) synthetic and degrading activity has been studied in steroid sensitive and resistant human leukemic cell lines. At least a two-fold increase in the levels of poly(A) polymerase activity was found in soluble, cytoplasmic extracts from the steroid sensitive human malignant T-cells of the MOLT3 line after 24-h treatment with dexamethasone. Longer exposure time of the cultured cells to the steroid resulted in a gradual decrease of the poly(A) polymerase activity level. Pretreatment of the cells with progesterone, a competitive inhibitor of dexamethasone, prevented the dexamethasone induced increase in the level of poly(A) polymerase activity, while progesterone alone had no effect on the enzyme level. In contrast, the levels of poly(A) nucleases activity remained constant following steroid treatment. In the steroid resistant human leukemic B-cell line Daudi no alterations in the poly(A) metabolizing enzymes could be measured in response to dexamethasone. Thus we suggest that poly(A) polymerase levels may be used to predict sensitivity of leukemic cells to glucocorticosteroid treatment. PMID- 3025531 TI - Histone gene stability during cellular senescence. AB - The extent to which human histone gene organization is conserved during the in vitro lifespan of human diploid fibroblast-like cells was determined by comparing the restriction patterns of a human H4 and an H3 histone gene from cells of various in vitro ages. No age related change in the organization of these two genes was detected. PMID- 3025530 TI - Bronchoscopic phototherapy with hematoporphyrin derivative for treatment of localized bronchogenic carcinoma: a 5-year experience. AB - Between December 1980 and April 1986 at our institution, 38 patients with cancer that involved the tracheobronchial tree (a total of 40 carcinomas) completed at least one course of hematoporphyrin derivative phototherapy. A complete response occurred in 13 patients (with 14 carcinomas). Eleven of these carcinomas did not recur during follow-up periods that ranged from 3 to 53 months. Three carcinomas recurred at 9, 12, and 35 months, respectively. For 26 carcinomas, the response was less than complete, and alternative therapy was necessary. The carcinomas in patients with a complete response were radiographically occult, were less than 3 cm2 in surface area, and appeared superficial at bronchoscopy. Our experience supports the use of hematoporphyrin derivative phototherapy as an alternative to surgical resection in carefully selected patients. PMID- 3025532 TI - [Incidence of hepatocarcinoma in our milieu in the last 10 years. Comparison with a previous study]. PMID- 3025533 TI - [Acute effects of guar gum after a test meal in type I diabetes. Insulin requirements with an artificial pancreas]. PMID- 3025534 TI - [Pneumonias associated with cytomegalovirus in immunodepressed patients]. PMID- 3025535 TI - Pituitary cyclic AMP and plasma hormone responses to epinephrine administration in vivo. AB - The present study was conducted to characterize the in vivo effects of epinephrine administration on levels of pituitary cyclic AMP and plasma hormones. Rats were injected with saline or epinephrine bitartrate (1 mg/kg lP) and sacrificed by decapitation 1, 5, 15, 30 or 60 min post-injection. Levels of pituitary cyclic AMP and plasma ACTH, beta-endorphin, beta-LPH, corticosterone and prolactin were determined by radioimmunoassays. The injection procedure itself was somewhat stressful as demonstrated by increased levels of plasma prolactin and ACTH 5 min following either saline or epinephrine injection. This "stress" response was rapid and short-lasting for the pituitary hormones. The response of the adrenal hormone, corticosterone, to saline injection was slower in onset and longer in duration. Pituitary cyclic AMP levels did not increase following saline injection. Epinephrine-injected animals displayed markedly elevated plasma levels of ACTH, beta-endorphin and beta-LPH at 15, 30 and 60 min as compared to control or saline-injected rats. In addition, levels of pituitary cyclic AMP were increased over 10 fold at these times. Levels of plasma prolactin, a stress-responsive hormone, were not significantly increased in epinephrine-injected animals as compared to saline-injected rats indicating that these later responses seem to be specific to epinephrine rather than to stress. PMID- 3025537 TI - Anatomical localization of corticotropin-releasing activity in the human brain. AB - Corticotropin-releasing activity (CRa) and arginine-vasopressin (AVP) content were measured in seven human hypothalami. The hypothalami were obtained from routine autopsy of patients suffering from no obvious neuroendocrinological abnormality. Twelve distinct hypothalamic areas were dissected in the frozen state and extracted in aqueous solution. CRa was measured by a bioassay measuring the aCTH released by rat pituitary cells in vitro, and vasopressin by direct radioimmunoassay. CRa was detectable in almost every area with the highest values in the supraoptic, paraventricular and infundibular (arcuate) areas. Vasopressin concentrations were maximum in the supraoptic nucleus, followed by the paraventricular and infundibular nuclei. We conclude that: hypothalami obtained from routine autopsy at a general hospital can be used for consistent CRa and vasopressin assay. Vasopressin and CRa are similarly distributed in man and in the rat. In both species, high CRa, which is not explained by AVP, is found in the paraventricular nucleus. The infundibular (arcuate) nucleus seems to display non AVP-dependent CRa much greater in the human than in the rat. PMID- 3025538 TI - Endogenous digitalis-like substance in pig left ventricle. AB - A crude fraction was isolated from pig heart left ventricle (150 g) homogenates after extraction of lipids, chromatographic separation and desalting. The extract contained an ionic content of 0.21, 0.27, 0.33 and 1.7 mM respectively for Mg2+, Ca2+, K+, and Na+. The albumin extract, used as a reference control, contained an ionic content of 0.88 and 2.1 mM respectively for K+ and Na+ and negligible amounts of Mg2+ and Ca2+. The isolated fraction exhibited digitalis-like properties in the inhibition of sarcolemmal Na+, K+-ATPase in a dose dependent manner, the displacement of [3H]-ouabain binding from membrane receptor sites and produced +ve inotropic response in isolated perfused heart in a dose dependent manner. The albumin extract tested in the same manner showed no digitalis-like properties. The ventricular fraction was unable to displace (-) 3H-DHA binding from membrane sites and its inotropic action was not blocked by propranolol. The data suggests that the fraction isolated from pig heart left ventricle contains a substance which has some properties like digitalis. PMID- 3025536 TI - Enhancing effects of angiotensin I on the vasopressin-stimulated water flow of toad bladder through increased cyclic AMP in mucosal cells. AB - The effects of angiotensins I and II on 10 mU/ml vasopressin-stimulated water flow across toad bladder were examined. Angiotensin I at concentrations of 10(-6) and 10(-7) M enhanced the water flow, but angiotensin II failed to do so at these concentrations. Angiotensin I had no effect on 5 mM cyclic AMP-stimulated water flow. After being preincubated for 30 min with angiotensin II, angiotensin I failed to have any stimulatory effect on vasopressin-stimulated water flow. At 10(-6) M angiotensin I significantly enhanced vasopressin-stimulated cyclic AMP content in bladder mucosal cells. These results indicate that angiotensin I enhances vasopressin-stimulated water flow by increasing cyclic AMP production in bladder cells and that angiotensin II may possibly interfere with angiotensin I in a competitive manner. PMID- 3025539 TI - Selective proliferation of brain kappa opiate receptors in spontaneously hypertensive rats. AB - The binding of tritiated ligands for various opiate receptor subtypes to brain membranes prepared from spontaneously hypertensive rats and normotensive Wistar Kyoto rats was determined. The density (Bmax) or the apparent dissociation constant (Kd) for the binding of the mu-ligand (naltrexone) and delta-ligand (Tyr D-Ser-Gly-Phe-Leu-Thr) to brain membranes of hypertensive and normotensive rats did not differ. However, the Bmax for the binding of kappa-ligand (ethylketocyclazocine, EKC) to brain membranes after the suppression of mu and delta-sites by 100 nM each of unlabeled D-Ala2-MePhe4-Gly-ol5-enkephalin and D Ala2-D-Leu5-enkephalin, respectively, was significantly greater in hypertensive rats compared to normotensive rats. The Kd values for the binding of 3H-EKC in the two groups did not differ. The binding of 3H-EKC in brain regions was in the order: hypothalamus greater than midbrain greater than striatum greater than cortex greater than pons + medulla. The increase in the binding of 3H-EKC in the brain of hypertensive rats compared to normotensive rats was due to increased binding in the hypothalamus and cortex. These results provide for the first time evidence of selective proliferation of kappa-opiate receptors in the brain of hypertensive rats, and suggest that brain kappa-opiate receptors may play an important role in the pathophysiology of hypertension. PMID- 3025540 TI - Adrenaline causes potassium influx in skeletal muscle and potassium efflux in cardiac muscle in rats: the role of Na/K ATPase. AB - Previous in vitro evidence suggests that adrenaline causes K influx in skeletal muscle by stimulating a ouabain sensitive Na/K ATPase membrane pump. However in rabbits, adrenaline induced hypokalaemia was not significantly altered by pretreatment with digoxin (50 micrograms/kg). Rats were infused with adrenaline or saline after being given a tracer dose of 42KCl. Adrenaline caused a highly significant uptake of 42K in skeletal muscle and a decrease in 42K uptake in ventricle. Rats were also studied after receiving a high dose of digoxin (1.4 mg/kg) which by itself produced a significant increase in plasma K, a decrease in plasma Na and a decreased uptake of 42K in ventricle and lung. These results suggest that adequate widespread Na/K ATPase inhibition had been achieved by this dose of digoxin but despite this, adrenaline still caused hypokalaemia and also still caused significant 42K tissue uptake by skeletal muscle. These results suggest that adrenaline causes K influx by skeletal muscle and K efflux by cardiac tissue. Furthermore, the former mechanism was not inhibited by pretreatment with digoxin. PMID- 3025541 TI - Angiotensin I converting enzyme inhibitors containing unnatural alpha-amino acid analogues of phenylalanine. AB - The activity of three angiotensin I converting enzyme (ACE) inhibitors with unique related structures was assessed in vitro and in vivo. The three compounds were (S)(-)-1,2,3,4-tetrahydro-2-(3-mercapto-1-oxopropyl)-3-isoquinoline carboxylic acid (EU-4865), 1,2,3,4-tetrahydro-2-(3-mercapto-1-oxopropyl)-1- isoquinolinecarboxylic acid (EU-4881), and (S)(-)-1,2,3,4-tetrahydro-1-(3 mercapto-1-oxopropyl)-2- quinolinecarboxylic acid (EU-5031). In vitro EU-4881 was a competitive inhibitor that lacked potency (IC50 = 1980 nM) against purified ACE. The other two compounds were equipotent (IC50 = 41 nM) against purified ACE but differed in their inhibition kinetics. EU-4865 (Ki = 38 nM) was a noncompetitive inhibitor, and EU-5031 (Ki = 6.9 nM) was a competitive inhibitor. Against caveolae membrane-bound ACE EU-4881 also lacked potency (IC50 = 2852 nM). In vivo in the conscious acute aortic coarctate (AAC) rat it also lacked potency, having an ED30 (oral dose decreasing blood pressure 30 mmHg) greater than 100 mg/kg. The activity of EU-4865 and EU-5031 in the caveolae membrane-bound ACE and AAC rat reflected their different Ki values rather than their similar IC50 values. In vitro, EU-4865 and EU-5031 had IC50 values of 19 and 6.7 nM, respectively, and in vivo, they had ED30 values of 52 and 1.1 mg/kg, respectively. These results suggest that ACE has a binding requirement for a carboxy-terminus, hydrophobic amino acid that is important for in vivo activity. PMID- 3025542 TI - Comparative effects of forskolin and isoproterenol on the cyclic AMP content of human adipocytes. AB - Alterations in adipocyte cyclic AMP concentrations in response to 100 microM forskolin and 10 microM isoproterenol over a 4 hour period were found to be similar; with each agent, a peak response was noted within 30 minutes. In general, the greater the magnitude of peak response, the more rapid the decline of cyclic AMP concentration during the ensuing 3 1/2 hours. Alpha-2 adrenergic activation, achieved with 10 microM clonidine or 10 microM epinephrine, substantially reduced the cyclic AMP concentrations in cells stimulated by 100 microM forskolin or 10 microM isoproterenol. Isoproterenol-stimulated cells appeared to be more sensitive to alpha adrenergic inhibition than did forskolin stimulated cells. Cells preincubated for 3 hours with 100 microM forskolin were markedly less responsive to a second exposure to the diterpine. Cells exposed to forskolin for 3 hours also had a reduced response when incubated with isoproterenol; thus, desensitization to forskolin appears to be heterologous. Forskolin desensitization did not appear to be dependent on cellular ATP depletion since cells mildly stimulated during preincubation were as severely desensitized as those adipocytes strongly stimulated. Maximum desensitization required a preincubation time of 1-2 hours with either isoproterenol or forskolin. PMID- 3025543 TI - Differential changes in atrial natriuretic peptide and vasopressin receptor bindings in kidney of spontaneously hypertensive rat. AB - To elucidate the role of atrial natriuretic peptide (ANP) and vasopressin (VP) in a hypertensive state, ANP and VP receptor bindings in spontaneously hypertensive rat (SHR) kidney were analyzed using the radiolabeled receptor assay (RRA) technique. Systolic blood pressure of SHR aged 12 weeks was statistically higher than that of age-matched Wistar Kyoto (WKY) rats. Maximum binding capacity (Bmax) of [125I]-ANP binding to the SHR kidney membrane preparations was statistically lower than that of WKY rats, but dissociation constant (Kd) was not significantly different. On the other hand, Bmax of [3H]-VP binding to the SHR kidney membrane preparations was statistically higher than that of WKY rats, but Kd were similar. Since the physiological action of ANP is natriuresis and VP is the most important antidiuretic hormone in mammalia, these opposite changes of ANP and VP receptor bindings in SHR kidney suggested that these peptides may play an important role in the pathophysiology of the hypertensive state, although it has not been confirmed as yet. PMID- 3025545 TI - [3H]muscimol binding sites increased in autopsied brains of chronic schizophrenics. AB - [3H]muscimol binding and glutamic acid decarboxylase (GAD) activity in the prefrontal cortex and caudate nucleus of autopsied brains from 19 chronic schizophrenics and 17 control subjects were investigated. In the schizophrenics, saturation analysis with varying concentrations of [3H]muscimol revealed an increase in the number of GABAA receptors, but there was no significant difference in the affinity. In addition, the enhancement of [3H]muscimol binding by diazepam was significantly greater in schizophrenics than in controls. GAD activity did not differ between controls and schizophrenics. The possibility that GABAergic mechanisms might play a role in case of chronic schizophrenia should be given further attention. PMID- 3025544 TI - Postnatal changes in enzyme activities of rat myocardial adenine nucleotide catabolic pathway. AB - Three catabolic enzymes, 5'-nucleotidase (5'NT), adenosine deaminase (ADA), purine nucleoside phosphorylase (PNP) and one anabolic enzyme, myokinase (MK) involved in adenine nucleotide (AN) metabolism were studied in myocardium from 4 to 105 day old rats. The specific enzyme activities (nmoles/min/mg protein) at day 4 were 35.3 for 5'NT, 28.4 for ADA, 43.3 for PNP, and 5 X 10(3) for MK. At day 7, 5'NT, activities rose to 450%; PNP and ADA 150%; and MK 120%; of the day 4 level. The activities of the three catabolic enzymes were elevated for one or two weeks then declined rapidly. By day 34, they were slightly above the adult values. MK activity displayed a different time course. It continued to increase slowly with age after the initial surge. Compared to the adult heart, the total activities of these catabolic enzymes in the one- to three-week-old heart were 30% to 220% higher. This transient elevation in AN catabolic enzyme activities may be related to active DNA synthesis and cell proliferation occurred in the rat myocardium during the same period. PMID- 3025546 TI - 16-Me cyprenorphine (RX 8008M): a potent opioid antagonist with some delta selectivity. AB - 16-Me cyprenorphine (RX 8008M) has been investigated in a number of isolated tissue preparations and found to be a pure opioid antagonist with Ke values at the delta, mu and kappa receptors of 0.73, 1.77 and 59.6 nM respectively. Comparisons of the mu, kappa and delta Ke values with a number of other antagonists in the mouse vas deferens have been made and show that the 16-Me substituent results in a marked enhancement of delta activity, making RX 8008M the most selective non-peptide delta antagonist available at the present time. PMID- 3025548 TI - [Dynamic roentgenometry and possibilities of prognosis in lung tumor response to treatment]. PMID- 3025547 TI - Interaction of [D-Ser2,Leu5]enkephalin-Thr6 (DSLET), a relatively selective delta ligand, with mu1 opioid binding sites. AB - Using binding approaches, we have confirmed the high selectivity of [D Ser2,Leu5]enkephalin-Thr6 (DSLET) to delta, as opposed to morphine-preferring (mu2) sites in rat brain. However, detailed experiments studies indicate that this ligand also labels mu1 sites with very high affinity. Saturation studies of 3H-DSLET binding reveal curvilinear plots. Treating tissue with naloxonazine to block mu1 sites, eliminates the higher affinity binding component. Competition studies of the other peptides against 3H-DSLET and 3H[D Ala2,MePhe4,Gly(ol)5]enkephalin (3H-DAMPGO) binding also implied high affinity binding of these peptides to mu1 sites. The ability of these peptides to interact with mu1 sites may help explain some of their pharmacological actions. PMID- 3025549 TI - Efficacy of Dexon microsutures in rat major abdominal vessel anastomosis. I: Application to heterotopic heart transplant. AB - A total of 52 syngeneic heterotopic heart transplants were performed using 9-0 Dexon microsutures to see if these anastomoses could be tolerable when challenged by the burden of transplanted heart and lung. Eighteen long-term surviving transplants were verified by reliable ECG, Doppler tracings, and direct palpation of the heart grafts. Animals sacrificed at monthly intervals beginning 3 to 6 months posttransplantation were free of thrombus, lung abscesses, and separation at the anastomoses. Even though foreign body reaction was still present up to 5 months posttransplantation, no actual sutures were present after 3 months. From these observations, Dexon microsuture is acceptable in organ transplantation procedures. PMID- 3025550 TI - Ouabain and low extracellular potassium inhibit PTH secretion from bovine parathyroid cells by a mechanism that does not involve increases in the cytosolic calcium concentration. AB - We have previously found that high extracellular calcium (Ca++) concentrations inhibit PTH release in association with a threefold to fourfold rise in cytosolic Ca++ concentration. Recent data have also shown that low extracellular potassium (K+) concentration or ouabain also inhibits PTH release to an extent comparable to that seen with high Ca++ and produce a marked rise in the intracellular sodium (Na+) content. These results suggested that low K+ and ouabain might modulate PTH release through increases in cytosolic Ca++ related to alterations in Na+-Ca++ exchange. In the present studies, we have examined further the mechanism(s) by which inhibition of the Na+-K+-ATPase regulates PTH release. Exposure of cells loaded with the Ca++-sensitive dye QUIN-2 to low K+ produced a 10% to 17% increase in cytosolic Ca++ at 0.5 to 1.0 mmol/L extracellular Ca++, which was statistically significant only at 0.75 mmol/L Ca++. In contrast, low K+ caused a statistically significant decrease in cytosolic Ca++ at 1.5 to 2 mmol/L Ca++, while ouabain lowered cytosolic Ca++ significantly by 23% to 46% at all Ca++ concentrations examined (0.5 to 2 mmol/L). Low K+ or ouabain had no effect on cellular levels of ATP or GTP or intracellular pH measured using the pH-sensitive dye BCECF [2', 7'-bis(carboxyethyl)-5,6-carboxyfluorescein]. The inhibition of secretion by low K+ or ouabain, unlike that due to high extracellular Ca++, was not reversed by TPA (12-O-tetradecanoyl phorbol 13-acetate), an activator of protein kinase C. Low K+ did produce a modest (30% to 40%) lowering of agonist stimulated but not basal cAMP content.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3025551 TI - Effects of diet restriction on insulin-sensitive phosphodiesterase in rat fat cells. AB - Effects of insulin on insulin-sensitive phosphodiesterase were investigated using fat cells from diet-restricted rats. The enzyme activities in diet-restricted rats were higher in case of 0 to 30 nmol/L insulin concentrations than in ad lib fed rats. In fat cells from the diet-restricted rats, the curves shifted to the left and half-maximum stimulation was obtained at 0.04 nmol/L, compared to that of 0.18 nmol/L in ad lib fed rats. Specific binding of insulin for fat cells from diet restricted and ad lib fed rats was 6.0% and 5.4%/2 X 10(5) cells, respectively. However when insulin binding was expressed per unit cell surface area, it was significantly increased in fat cells from diet-restricted rats compared with that from control rats. These results suggest that the insulin effector system related to the phosphodiesterase activation is improved in fat cells from diet-restricted rats, in particular, sensitivity to insulin. These increases in sensitivity to the phosphodiesterase are possibly due to improvements of insulin receptor binding. PMID- 3025552 TI - Free amino acid levels in muscle and liver of a patient with glucagonoma syndrome. AB - This study set out to measure the amino acid concentrations in the femoral artery, femoral vein, hepatic vein, muscular and hepatic tissue, and urine of a patient with the glucagonoma syndrome. The total plasma amino acid concentration was severely reduced on admission (737 mumol/L, 26% of normal), with only a slight increase during intravenous administration of 200 g of amino acids per day. The total intracellular amino acid levels in the muscle were 86%, and those of the liver were 47% of the normal range. Only 0.62% of the amino acids administered were found in the urine. Arteriovenous amino acid concentration differences across the muscle and splanchnic tissue indicated the release of amino acids (mainly glutamine, glycine, and alanine) from the muscle and the absorption of amino acids by the splanchnic bed. This study shows that the infusion of a high amount of amino acids cannot increase the subnormal plasma AA levels of patients with the glucagonoma syndrome. The low total plasma AA levels are paralleled by decreased intracellular free amino acid levels in the muscle and liver. PMID- 3025553 TI - Regulation of transcription of the chromosomal dnaA gene of Escherichia coli. AB - By comparative S1 analysis we investigated the in vivo regulation of transcription of the chromosomal dnaA gene coding for a protein essential for the initiation of replication at the chromosomal origin. Inactivation of the protein in dnaA mutants results in derepression, whereas excess DnaA protein (presence of a DnaA overproducing plasmid) leads to repression of dnaA transcription. Both dnaA promoters are subject to autoregulation allowing modulation of transcriptional efficiency by at least 20-fold. Increasing the number of oriC sequences (number of DnaA binding sites) in the cell by introducing oriC plasmids leads to a derepression of transcription. Autoregulation and binding to oriC suggest that the DnaA protein exerts a major role in the regulation of the frequency of initiation at oriC. The efficiency of transcription of the dnaA2 promoter is reduced in the absence of dam methylation, which is involved in the regulation of oriC replication. PMID- 3025554 TI - Repeated sequences with unusual properties from the divergent region of Crithidia oncopelti maxicircle kinetoplast DNA. AB - The occurrence of bacterial promoter-like sequences was shown in the divergent region of Crithidia oncopelti maxicircular kinetoplast DNA. A promoter-cloning vector based on the neomycin phosphotransferase II (NPT II) gene was used to localize promoters in three homologous blocks of repeated sequences. The elements found displayed greater promoter strength in Escherichia coli than the normal promoter of the NPT II gene. Sequences of three promoter-containing inserts from different blocks were determined and typical prokaryotic promoter sequences were localized. PMID- 3025555 TI - The carboxy-terminal region of haemolysin 2001 is required for secretion of the toxin from Escherichia coli. AB - As a first step in the detailed analysis of the mechanism of secretion of haemolysin, we sought to identify sequences or domains within haemolysin A (HlyA) that are essential for its secretion. For this purpose we examined the properties of a deletion and Tn5 insertions into the region of the HlyA gene encoding the C terminal part of the protein, since both of these are relatively simple to generate. We showed that removal of 27 amino acids from the C-terminus of HlyA is sufficient to inhibit secretion drastically, although the residual polypeptide is still haemolytically active. Cellular fractionation studies showed that haemolytic activity does not accumulate in large amounts within the periplasmic space during normal secretion. More significantly, activity does not appear to accumulate within this compartment when the export functions hlyB and hlyD are removed. These results are consistent with a mechanism in which interaction of the C-terminus of HlyA with the secretion machinery, located in the inner membrane, is followed by direct transfer of haemolysin to the medium. PMID- 3025557 TI - T-even type phages can change their host range by recombination with gene 34 (tail fibre) or gene 23 (head). AB - T-even type phages recognize their cellular receptors with the tip of their long tail fibres. The gene products involved in receptor recognition are proteins 37 and 38. While screening libraries of phage K3 with a probe of gene 38 from phage T2, a class of weakly hybridizing clones was found in addition to the expected clones of gene 38 of K3. One of these clones was identified as being from gene 23 of the phage which codes for the major head subunit; another clone originated from gene 34, which codes for the proximal half of the long tail fibres. Neither gene product 23 nor 34 is involved in receptor recognition. Phages can recombine with the DNA of the gene 23 and gene 34 clones and change the host range. PMID- 3025556 TI - A new cell division operon in Escherichia coli. AB - At 76 min on the E. coli genetic map there is a cluster of genes affecting essential cellular functions, including the heat shock response and cell division. A combination of in-vivo and in-vitro genetic analysis of cell division mutants suggests that the cell division gene fts E is the second gene in a 3 gene operon. A cold-sensitive mutant, defective in the third gene, is also unable to divide at the restrictive temperature, and we designate this new cell division gene fts X. Another cell division gene, fts S, is very close to, but distinct from, the 3 genes of the operon. The fts E product is a 24.5 Kd polypeptide which shows strong homology with a small group of proteins involved in transport. Both the fts E product and the protein coded by the first gene (fts Y) in the operon have a sequence motif found in a wide range of heterogeneous proteins, including the Ras proteins of yeast. This common domain is indicative of a nucleotide binding site. PMID- 3025558 TI - An upstream signal is required for in vitro transcription of Neurospora 5S RNA genes. AB - The DNA sequences upstream of the 5S RNA genes in Neurospora crassa are largely different from one another, but share a short consensus sequence located in the segment 29 to 26 nucleotides preceding the transcribed region. Differences among flanking sequences do not appear to affect transcription. Deletion analysis indicates, however, that a DNA segment including the conserved "TATA box" is required for in vitro transcription of Neurospora 5S RNA genes. PMID- 3025559 TI - Characterization of the gene products produced in minicells by pSM1, a derivative of R100. AB - At least ten polypeptides larger than 6 kilodaltons (K) are produced in minicells from the miniplasmid pSM1 in vivo. pSM1 (5804 bp) is a small derivative of the drug resistance plasmid R100 (ca. 90 kb) and carries the R100 essential replication region as well as some non-essential functions. Cloned restriction fragments of pSM1 and plasmids with deletions within pSM1 sequences were used to assign eight of the ten observed polypeptides to specific coding regions of pSM1. Two of these polypeptides were identified as RepA1 and RepA2, proteins encoded by the essential replication region of pSM1/R100. The nucleotide sequence consisting of 885 bp outside the essential replication region is presented here. This sequence contains an open reading frame, orf4, for a protein 22.9 K in size, and one of the pSM1-encoded polypeptides was identified as the orf4 gene product. Five additional polypeptides were shown to be the products of other open reading frames mapping outside the essential replication region. Specific functions have been assigned to four of these polypeptides and tentatively to the fifth. PMID- 3025560 TI - Physical and genetic characterisation of the gene cluster for the antibiotic actinorhodin in Streptomyces coelicolor A3(2). AB - We determined the physical and transcriptional organisation of the set of previously cloned biosynthetic genes involved in the production of the polyketide antibiotic actinorhodin by Streptomyces coelicolor A3(2). Complementation and mutational cloning analyses (in part using new phi C31 phage vectors incorporating a transcriptional terminator to block transcription from vector promoters into the cloned DNA) indicate that all the biosynthetic genes, including at least one regulatory (activator) gene, are clustered in a chromosomal region of about 26 kb. The genes are organised in at least four separate transcription units, ranging in size from 1 kb for the class III gene, to a polycistronic transcript of at least 5 kb for the class I, VII and IV genes. Indirect evidence shows that resistance to actinorhodin is also determined by the cloned DNA. PMID- 3025561 TI - Genetic characterization and isolation of the Saccharomyces cerevisiae gene coding for uridine monophosphokinase. AB - We selected a 5-fluorouracil-resistant, thermosensitive mutant of the uridine monophosphokinase step in Saccharomyces cerevisiae. The mutant displays very weak thermolabile uridine monophosphokinase activity and wild-type uridine diphosphokinase activity. Growth of the mutant at the non-permissive temperature causes immediate reduction of pyrimidine triphosphate pools to 10% of the wild type level as well as significantly lowering total RNA and protein synthesis. These conditions also provoke derepression of the first gene of the pathway, URA2, at both the levels of enzymatic activity and transcription. The mutation segregates independently of all known genes of the pyrimidine biosynthetic pathway. The corresponding gene has been isolated on a 4.8 kb fragment by complementation of the mutant phenotype. The new gene, named URA6, codes for a 2.2 kb polyadenylated messenger RNA, exists in a single copy per haploid genome, and was mapped to the centromere of chromosome XI. PMID- 3025562 TI - Complete nucleotide sequence of the low copy number plasmid pRAT11 and replication control by the RepA protein in Bacillus subtilis. AB - The 2.6 kb kanamycin-resistant (Kmr) plasmid, pRAT11, was constructed using both the replication determinant (repA) region of the 10.8 kb tetracycline-resistant (Tcr) low copy number plasmid pTB52 and another fragment (0.9 kb) that contained solely the Kmr gene of pUB110. The complete nucleotide sequence of this plasmid was determined. The repA region contained a large open reading frame encoding RepA protein (396 amino acid residues). In vitro transcription and translation of the repA gene were confirmed. RepA protein was shown to be indispensable for plasmid replication, and acted in trans on DNA. The part of the repA gene encoding the specific recognition region of the RepA protein was located and contained 3.5 direct repeats of 24 bp (GGTTTCAAAAATGAAACGGTGGAG). Upstream and downstream of the direct repeats were the recognition sequence (TTATCCACA) of the Escherichia coli DnaA protein and an AT-rich region, respectively. The replication control mechanism of the low copy number Bacillus plasmid is discussed. PMID- 3025564 TI - Ultrastructural study of biochemically modulated ADCC in HSV-1 infected and uninfected Chang liver cells. AB - The effect of cytochalasin B, ouabain and 25-OH cholesterol on specific lysis due to antibody dependent cellular cytotoxicity (ADCC) in allogeneic and xenogeneic systems was studied using Herpes simplex I infected Chang liver cells. Cytochalasin B reduced both cytotoxicity and lymphocyte/target (LT) binding in the allogeneic system whereas cytotoxicity but not LT binding was reduced in the xenogeneic system. Ouabain inhibited ADCC in both systems as well as LT binding in the allogeneic system; however, binding in the xenogeneic system was not significantly reduced. The 25-OH cholesterol produced a marked decrease in ADCC in both systems but had no significant effect on LT binding in either system. The biochemical and ultrastructural data suggest that the modulators act at different stages in the ADCC response and that there may be more than one mechanism of ADCC to handle different types of target antigens. PMID- 3025563 TI - Molecular cloning and sequence of the Bacillus stearothermophilus translational initiation factor IF2 gene. AB - The structural gene for the Bacillus stearothermophilus initiation factor IF2 was localized to a 6 kb HindIII restriction fragment by cross-hybridization with the SstI-SmaI fragment of the Escherichia coli infB gene. This fragment corresponds to the central region of the molecule containing the GTP-binding domain which is homologous in E. coli IF2, EF-Tu, EF-G and the human ras1 oncogene protein. After cloning into pACYC177, the HindIII fragment was further analysed by restriction mapping and cross-hybridization. A smaller (2.2 kb) SphI-HindIII fragment, which showed cross-hybridization, was subcloned into M13 phage and sequenced by the dideoxy chain-terminating method. This fragment was found to contain the entire IF2 gene except for the region coding for the N-terminus. This remaining region, coding for 45 amino acids, was located by homologous hybridization on an overlapping ClaI-SstI fragment which was also subcloned and sequenced. Overall, the B. stearothermophilus IF2 gene codes for a protein of 742 amino acids (Mr = 82,043) whose primary sequence displays extensive homology with the C-terminal two-thirds (but little or no homology with the N-terminal one-third) of the corresponding E. coli IF2 molecule. When cloned into an expression vector under the control of the lambda PL promoter, the B. stearothermophilus IF2 gene, reconstituted by ligation of the two separately cloned pieces, could be expressed at high levels in E. coli cells. PMID- 3025565 TI - Vascular tracers alter hemodynamics and airway pressure in anesthetized sheep. AB - The technique of vascular labeling was developed to mark sites of increased microvascular permeability. We used the vascular labeling technique in anesthetized sheep and found that hemodynamics and airway pressure were adversely affected by intraarterial infusions of two vascular tracers. Monastral blue (nine sheep) immediately caused systemic arterial hypotension, pulmonary arterial hypertension, and bronchoconstriction. All three physiological responses were partially blocked by a cyclooxygenase inhibitor (indomethacin) but not by an H1 antihistamine (chlorpheniramine). Colloidal gold (nine sheep) caused immediate, but less dramatic, pulmonary arterial hypertension which was not attenuated by the blocking agents. We conclude that these two vascular tracers caused detrimental physiological side effects in sheep at the usual doses used to label injured microvessels in other species. PMID- 3025566 TI - Monoclonal antibodies against Chlamydia psittaci. AB - Five monoclonal antibodies were prepared against Chlamydia (C.) psittaci strain Pigeon-1041 isolated from a feral pigeon in Sapporo. Reactions of these antibodies to chlamydiae were examined using five strains of C. psittaci and two strains of C. trachomatis in an enzyme-linked immunosorbent assay, microimmunofluorescent test and complement fixation test. The antibodies were divided into two groups: three genus-specific (A2, D2, and I21) and two strain specific (F2 and H9) antibodies. The antigenic determinant site of A2 was KIO4 sensitive, but those of D2, F2, and H9 were not affected greatly by KIO4 treatment. Nine C. psittaci strains from feral pigeons and 16 strains from budgerigars were classified into three groups and four groups, respectively, by reaction patterns against the monoclonal antibodies. PMID- 3025567 TI - Protective effect of acidic mannan fraction of bakers' yeast on experimental candidiasis in mice. AB - An acidic fraction of bakers' yeast mannan, WAM025, showed a significant protective effect against Candida albicans infection in mice, but a neutral fraction of the same bakers' yeast mannan, WNM, did not exhibit this effect. Moreover, pretreatment with WAM025 resulted in a marked reduction of proliferation of C. albicans cells in the organs of the infected mice. We investigated the stimulative effect of these mannan fractions on the function of mouse peritoneal phagocytes, and found that mice administered WAM025 showed a greater increase in the number of peritoneal exudate cells, macrophages and polymorphonuclear leucocytes (PMN), than the mice treated with WNM, especially in the proportion of PMN. Peritoneal phagocytes, PMN and macrophages obtained from WAM025-treated mice showed marked candidacidal activity. Of the phagocytes, PMN were responsible for the larger part of the candidacidal activity. The myeloperoxidase activities of PMN and macrophages in WAM025-treated PEC were greater than in untreated macrophages. The myeloperoxidase activity of WAM025 treated PMN was significantly greater than that of WAM025-treated macrophages. This activity paralleled the active oxygen-releasing activity of the phagocytes. On the other hand, the phagocytic activity of phagocytes from mice administered WNM or WAM025 for C. albicans cells was identical to that of untreated phagocytes. WAM025 seems to cause enhance elimination of the pathogen from mice, by increasing the number and candidacidal activity of phagocytic cells. PMID- 3025568 TI - A new concept in the pathogenesis of drug-induced psoriasis. AB - Since drug-induced psoriasis is triggered by a known agent, studies of this type of psoriasis may provide clues to understanding the pathogenesis of the disease. Analysis of the reported cases of drug-induced psoriasis lead us to the postulation of a new concept of this disease. According to this concept drug induced psoriasis is an auto-immune disease of the beta receptors of the skin. The initial and central event is the alteration of the immunological properties of the receptor due to an interaction with the drug. This leads further to the activation of an immunological cascade that results in a reduction in the functional activity of the receptors and through a known pathway to the development of the clinical manifestation of the disease. PMID- 3025569 TI - The frequency of non-A, non-B hepatitis in acute and chronic liver disease. PMID- 3025570 TI - The hepatic side-effects of drugs. PMID- 3025571 TI - Comparison between Premarin and ethinyl oestradiol. PMID- 3025572 TI - [Asbestos and talc: determination of the fibrous mineral particle content in commercial pulverized talc using combined optical microscope technics]. PMID- 3025573 TI - Drugs for hypertension. PMID- 3025574 TI - Fish oil for the heart. PMID- 3025575 TI - [Activity of the cyclic adenosine monophosphate phosphodiesterase of the blood lymphocytes in patients with a Lamblia infection susceptible and resistant to lambliacides]. PMID- 3025576 TI - [Corticotropin releasing factor (CRF) stimulation test in asthmatic patients with different steroid medications]. PMID- 3025577 TI - [Oncogenes and neoplasms in children: solid tumors. I]. PMID- 3025579 TI - [Hemoperitoneum: the initial symptom of hepatoblastoma in a child operated on with success]. PMID- 3025578 TI - [Silver-Russell syndrome. A possible example of genetic heterogenicity]. PMID- 3025580 TI - How do the elderly fare in english society? Care of the aged in Oxford, England. Part I. PMID- 3025581 TI - Neurology of sex steroids and oral contraceptives. AB - Under normal circumstances, sex steroids interact with diverse neural substrates to modulate a host of activities essential to the preservation of the individual and the species. In addition, sex hormones play an important role in various human neurologic conditions including strokes, migraine, certain movement disorders and peripheral neuropathies, and possibly even the behavior of CNS neoplasms. PMID- 3025582 TI - Neuropeptides in Alzheimer's disease. Clinical implications. AB - Since the isolation and sequencing of the enkephalins in the mid-1970s, there has been an explosion of knowledge concerning putative peptide neurotransmitters in the brain. This article focuses on selected developments in neuropeptide research that may have bearing on our understanding of and clinical approach to the dementing degenerative disorder Alzheimer's disease (AD), a cardinal feature of which is impairment of memory. PMID- 3025583 TI - Neuropeptides and seizures. AB - There are four lines of evidence for or against a role of neuropeptides in epilepsy: Administration of a variety of opiate agonists into the ventricles or brain of animals produces a constellation of electrical and behavioral changes, seemingly receptor-specific, both sensitive to the specific opiate antagonist naloxone as well as certain anticonvulsant drugs. The primary reservation concerning these data in terms of their relevance to epilepsy regards the fact that the peptides are exogenously administered in relatively high doses. Hence, these data may reflect neurotoxic effects of peptides rather than physiologic function. A variety of opiate agonists are anticonvulsant and naloxone shortens the postictal state in some experimental seizure models. One could attempt to reconcile these data with those in No. 1 by hypothesizing that the spikes and behavioral changes examined in the latter experimental parodynes represented a sort of isolated model of the postictal state. Naloxone has little effect in clinical epilepsy. These data are far from conclusive for two reasons. First, few patients have been studied. Second, because of the issue of opiate receptor heterogeneity and the high doses of naloxone needed experimentally to block non mu opiate effects, the doses of naloxone used clinically to date are too low to rule out possible delta- or epsilon-mediated effects. The negative clinical data are illustrative of the dangers and difficulties of extrapolating data generated in animal models of seizures to the human condition. ACTH, a peptide that is derived from the same precursor molecule as beta-endorphin, is clearly an effective anticonvulsant in certain childhood seizure states. However, whether this is due to a direct or indirect (that is, cortisol) effect on brain is far from clear. Paradoxically, in contradistinction to other data concerning pro- and anticonvulsant properties of various opioid peptides, there is no animal model of infantile spasms to help resolve this important question. PMID- 3025584 TI - Neuroendocrine aspects of pineal tumors. AB - The evaluation and treatment of pineal region tumors has changed dramatically in the past decade. New imaging techniques result in earlier diagnosis. Surgical techniques now permit removal of benign tumors (about one third of cases). Pathologic diagnosis is obtained for the remainder, and postoperative therapy can be planned rationally. Germ cell tumors pose a particularly difficult problem for the pathologist because they often contain mixed germ cell elements. Biologic markers, beta HCG and AFP, aid diagnosis and can be monitored in serum and CSF to assess response to treatment. Only boys develop precocious puberty with pineal tumors. What appears to be puberty is actually pseudoprecocious puberty, due to ectopic production of HCG by their neoplasms (choriocarcinomas or germinomas with syncytiotrophoblastic giant cells). HCG can stimulate tests to produce testosterone, but FSH is necessary (together with LH or HCG) to stimulate ovaries to produce estrogen. Diabetes insipidus with pineal tumors is usually due to spread to the hypothalamus by ventricular seeding, but we report a case of aqueductal stenosis with diabetes insipidus resulting from a massively dilated third ventricle. Rarely, hydrocephalus may trigger true precocious puberty, a syndrome easily differentiated from pseudoprecocious puberty by endocrinologic tests. There are no biologic markers to diagnose pineal parenchymal tumors. Elevation of melatonin in plasma or CSF and increased tumor biosynthetic activity has been reported in isolated cases, but the range of melatonin values in normals is very wide, and melatonin levels do not correlate with specific pathologic tumor types. Both parenchymal and nonparenchymal tumors may increase melatonin nonspecifically by interfering with regulatory mechanisms of the normal pineal gland. PMID- 3025586 TI - Sites of P element insertion and structures of P element deletions in the 5' region of Drosophila melanogaster RpII215. AB - Several P element insertion and deletion mutations near the 5' end of Drosophila melanogaster RpII215 have been examined by nucleotide sequencing. Two different sites of P element insertion, approximately 90 nucleotides apart, have been detected in this region of the gene. Therefore, including an additional site of P element insertion within the coding region, there are at least three distinct sites of P element insertion at RpII215. Both 5' sites are within a noncoding portion of transcribed sequences. The sequences of four revertants of one P element insertion mutation (D50) indicate that the P element is either precisely deleted or internally deleted to restore RpII215 activity. Partial internal deletions of the P element result in different RpII215 activity levels, which appear to depend on the specific sequences that remain after excision. PMID- 3025585 TI - Transcriptional analysis of Ty1 deletion and inversion derivatives at CYC7. AB - One class of Ty insertion mutation in Saccharomyces cerevisiae activates expression of adjacent structural genes. The CYC7-H2 mutation, in which a Ty1 element is inserted 5' to the iso-2-cytochrome c coding region of CYC7, causes a 20-fold increase in CYC7 expression. Deletion analysis of CYC7-H2 has shown that distal regions of the Ty1 element are not essential for the transcriptional activation at CYC7. In this report, we have analyzed Ty1 and CYC7 RNA from two CYC7-H2 deletion derivative genes to determine whether a direct correlation exists between transcription of Ty1 and transcription of the adjacent gene. Assuming that all Ty1 elements in the genome are transcribed equally, amounts of CYC7-H2 deletion derivative Ty1 RNA were found to be at least fivefold lower than the amount estimated for the average Ty1 element. These same Ty1 deletion derivatives caused a 20-fold increase in adjacent CYC7 expression. This finding suggests that the mechanism by which Ty1 activates adjacent gene expression does not require normal levels of Ty1 transcription. Two inversion derivatives of the CYC7-H2 Ty1 have also been analyzed. These derivatives did not produce any iso-2 cytochrome c or any normal CYC7 mRNA. Instead they were found to produce a Tyl CYC7 fusion RNA. Consistent with our findings on CYC7-H2 Ty1 transcription, the amount of the fusion RNA was very low. In addition, the Ty1 inversion derivatives produced a new RNA that mapped to sequences upstream from the inverted Ty1 segment. Similar to Ty1 insertions that activate transcription, the new RNA was found to be transcribed away from Ty1. PMID- 3025587 TI - Expression of the Saccharomyces cerevisiae inositol-1-phosphate synthase (INO1) gene is regulated by factors that affect phospholipid synthesis. AB - The INO1 gene of Saccharomyces cerevisiae encodes the regulated enzyme inositol-1 phosphate synthase, which catalyzes the first committed step in the synthesis of inositol-containing phospholipids. The expression of this gene was analyzed under conditions known to regulate phospholipid synthesis. RNA blot hybridization with a genomic clone for INO1 detected two RNA species of 1.8 and 0.6 kb. The abundance of the 1.8-kb RNA was greatly decreased when the cells were grown in the presence of the phospholipid precursor inositol, as was the enzyme activity of the synthase. Complementation analysis showed that this transcript encoded the INO1 gene product. The level of INO1 RNA was repressed 12-fold when the cells were grown in medium containing inositol, and it was repressed 33-fold when the cells were grown in the presence of inositol and choline together. The INO1 transcript was present at a very low level in cells containing mutations (ino2 and ino4) in regulatory genes unlinked to INO1 that result in inositol auxotrophy. The transcript was constitutively overproduced in cells containing a mutation (opi1) that causes constitutive expression of inositol-1-phosphate synthase and results in excretion of inositol. The expression of INO1 RNA was also examined in cells containing a mutation (cho2) affecting the synthesis of phosphatidylcholine. In contrast to what was observed in wild-type cells, growth of cho2 cells in medium containing inositol did not result in a significant decrease in INO1 RNA abundance. Inositol and choline together were required for repression of the INO1 transcript in these cells, providing evidence for a regulatory link between the synthesis of inositol- and choline-containing lipids. The level of the 0.6-kb RNA was affected, although to a lesser degree, by many of the same factors that influence INO1 expression. PMID- 3025588 TI - Multiple sequence elements are required for maximal in vitro transcription of a human histone H2B gene. AB - As part of our studies on the cell cycle regulation of human histone gene expression, we examined the elements governing transcription of a human histone H2B gene in nuclear extracts derived from human HeLa cells. Circular templates were transcribed at 5- to 10-fold higher levels than were linear templates. A series of deletion, linker-substitution, and point mutants defined cis-acting promoter sequences that were recognized in nuclear extracts. These sequences extended from 118 to 21 base pairs 5' to the transcription initiation site. Elements recognized included (from 5' to 3') a series of direct repeats, a CCAAT homology, a human histone-specific hexamer, an H2B consensus element, and a TATA box. Sequence elements 5' to the hexamer were required for its function. In contrast, the H2B consensus element could function independently of more-5' promoter elements and in turn was essential for the function of upstream elements. An interesting feature of this consensus is that its core octanucleotide (ATTTGCAT) is found in several nonhistone genes. By comparison with functional elements in an H4 promoter, we infer that a combinatorial interaction of general and gene-specific factors may contribute to the S-phase elevation of H2B transcription. PMID- 3025590 TI - 5'-flanking sequence required for regulated expression of a muscle-specific Drosophila melanogaster actin gene. AB - We have functionally tested derivatives of a muscle-specific Drosophila melanogaster actin gene in which 5'-flanking sequences have been deleted or rearranged. From our results we conclude that approximately 1,000 nucleotides of 5'-flanking sequence are required for wild-type levels of mRNA accumulation during flight muscle development. Derivatives having 875 or 865 nucleotides of upstream sequence could be expressed normally, but were prone to influence by flanking foreign DNA sequences. Derivatives retaining 600 or fewer nucleotides of flanking DNA did not direct detectable levels of mRNA accumulation. The sequence residing between -919 and -640 could be inverted and yet retain normal function. Deletion of this sequence reduced mRNA accumulation markedly, but did not affect its spatial localization, suggesting that elements which confer tissue specificity reside close to the point of transcription initiation. PMID- 3025589 TI - Differential response to retinoic acid of Syrian hamster embryo fibroblasts expressing v-src or v-Ha-ras oncogenes. AB - It has been shown that treatment of many but not all tumor cell lines with retinoids affects cell proliferation and expression of the transformed phenotype. To determine whether the response of the tumor cell to retinoids is influenced by specific oncogenes activated in the cell, we studied the action of these agents in the immortal, nontumorigenic Syrian hamster embryo cell lines DES-4 and 10W transfected with either v-Ha-ras or v-src oncogenes. In this paper we show that in transformed DES-4 cells expressing v-src, retinoic acid inhibited anchorage independent growth, reduced saturation density, and inhibited the induction of ornithine decarboxylase by the phorbol ester 12-O-tetradecanoylphorbol-13 acetate. In contrast, retinoic acid enhances the expression of the transformed phenotype in DES-4-derived cells that express v-Ha-ras. In these cells retinoic acid increases the number and the average size of colonies formed in soft agar. Moreover, retinoic acid enhances ornithine decarboxylase activity and acts in a synergistic fashion with 12-O-tetradecanoylphorbol-13-acetate. These results indicate that oncogenes activated in cells can indeed influence the response of cells to retinoids. Retinoic acid does not appear to alter the levels of pp60src or p21ras proteins in these cells, suggesting that retinoic acid does not affect the synthesis of these oncogene products. Furthermore, retinoic acid does not affect the protein kinase activity of pp60src. Transformed cell lines derived from 10W cells responded differently, indicating that the presence of a specific oncogene is not the only factor determining the response to retinoids. Possible mechanisms by which retinoic acid may interfere with the expression of the oncogene products are discussed. PMID- 3025591 TI - Repair of heteroduplex plasmid DNA after transformation into Saccharomyces cerevisiae. AB - Purified heteroduplex plasmid DNAs containing 8- or 12-base-pair insertion mismatches or AC or CT substitution mismatches were used to transform Saccharomyces cerevisiae. Two insertion mismatches, separated by 943 base pairs, were repaired independently of each other at least 55% of the time. This suggested that repair tracts were frequently shorter than 1 kilobase. The two insertion mismatches were repaired with different efficiencies. Comparison of the repair efficiency of one mismatched site with or without an adjacent mismatch suggests that mismatches promote their own repair and can influence the repair of neighboring mismatches. When two different plasmids containing single-insertion mismatches were transformed into S. cerevisiae cells, a slight preference towards insertion was detected among repair products of one of the two plasmids, while no repair preference was detected among transformants with the second plasmid. PMID- 3025592 TI - Activated v-myc and v-ras oncogenes do not transform normal human lymphocytes. AB - Activated v-myc (pSV v-myc) and v-Ha-ras (GT10) oncogenes were introduced into normal human lymphocytes, NIH 3T3 fibroblasts, B-lymphoblastoid cells, and human epithelial cells, using a reconstituted Sendai virus envelope-mediated gene transfer technique. Efficient transfer of the plasmid in each cell type was demonstrable within 1.5 h of transfection by Southern blotting of extrachromosomal DNA extracts, which unexpectedly revealed that v-myc plasmid DNA was unstable in normal lymphocytes but not in the other cell types. The v-myc plasmid was stabilized when cotransfected into lymphocytes together with v-Ha ras. The transfected v-Ha-ras plasmid was stable in all the cell types tested. v myc plasmid expression was clearly detectable by 5 h in all cell types except human lymphocytes. Lymphocytes expressed v-myc when transfected together with v Ha-ras. Transfected ras oncogene was efficiently expressed in all the cell types tested. Expression of the transfected genes increased at 24 and 48 h after transfection. Even though plasmid stability and expression were achieved in myc ras-cotransfected lymphocytes, no effects on cellular DNA synthesis or immortalization were observed, in contrast to efficient transformation of NIH 3T3 fibroblasts by the same procedure. Our data suggest that efficient expression of transfected myc and ras oncogenes in normal quiescent human lymphocytes is not sufficient for the induction of cell growth and immortalization. PMID- 3025593 TI - Isolation and characterization of expressible cDNA clones encoding the M1 and M2 subunits of mouse ribonucleotide reductase. AB - Mammalian ribonucleotide reductase consists of two nonidentical subunits, proteins M1 and M2, which are differentially regulated during the cell cycle. We have isolated expressible cDNA clones of both subunits from an Okayama-Berg cDNA library made with mRNA from hydroxyurea-resistant, M2 protein-overproducing mouse TA3 cells. Expression of M2 protein could be demonstrated by electron paramagnetic resonance spectroscopy after transfection of COS-7 monkey cells with the plasmid. Electrophoresis and blot analyses of the parent and hydroxyurea resistant TA3 mRNA revealed two M2 transcripts, a major one of 2.1 kilobases and a minor one of about 1.6 kilobases. Restriction endonuclease mapping of the corresponding cDNAs indicated that the two mRNAs differed only in the length of the 3' untranslated ends. By contrast, there was only one mRNA corresponding to the M1 protein, and its mobility corresponded to about 3.1 kilobases. The hydroxyurea-resistant TA3 cells contained a 50- to 100-fold excess of the M2 mRNAs over that of the parent cells and a 10-fold excess of the M1 mRNA. However, a Southern blot analysis of the corresponding genomic DNA sequences showed that the M2 gene was amplified fivefold but the M1 gene was still single copy. The complete nucleotide sequence of the 2,111-base-pair-long M2 cDNA revealed an open reading frame coding for 390 amino acids, which corresponds to a molecular weight of 45,100. The mouse M2 protein sequence was quite homologous to the equivalent protein in the clam Spisula solidissima, while the homology to the smaller subunits of Epstein-Barr virus, herpes simplex virus type 2, and Escherichia coli ribonucleotide reductases were less pronounced. PMID- 3025594 TI - Pyrimidine dimers block simian virus 40 replication forks. AB - UV light produces lesions, predominantly pyrimidine dimers, which inhibit DNA replication in mammalian cells. The mechanism of inhibition is controversial: is synthesis of a daughter strand halted at a lesion while the replication fork moves on and reinitiates downstream, or is fork progression itself blocked for some time at the site of a lesion? We directly addressed this question by using electron microscopy to examine the distances of replication forks from the origin in unirradiated and UV-irradiated simian virus 40 chromosomes. If UV lesions block replication fork progression, the forks should be asymmetrically located in a large fraction of the irradiated molecules; if replication forks move rapidly past lesions, the forks should be symmetrically located. A large fraction of the simian virus 40 replication forks in irradiated molecules were asymmetrically located, demonstrating that UV lesions present at the frequency of pyrimidine dimers block replication forks. As a mechanism for this fork blockage, we propose that polymerization of the leading strand makes a significant contribution to the energetics of fork movement, so any lesion in the template for the leading strand which blocks polymerization should also block fork movement. PMID- 3025595 TI - Identification of separate domains in the adenovirus E1A gene for immortalization activity and the activation of virus early genes. AB - The transformation and early adenovirus gene transactivation functions of the E1A region were analyzed with deletion and point mutations. Deletion of amino acids from position 86 through 120 had little effect on the lytic or transforming functions of the E1A products, while deletion of amino acids from position 121 through 150 significantly impaired both functions. The sensitivity of the transformation function to alterations in the region from amino acid position 121 to 150 was further indicated by the impairment of transforming activity resulting from single amino acid substitutions at positions 124 and 135. Interestingly, conversion of a cysteine residue at position 124 to glycine severely impaired the transformation function without affecting the early adenovirus gene activating functions. Single amino acid substitutions in a different region of the E1A gene had the converse effect. All the mutants produced polypeptides of sufficient stability to be detected by Western immunoblot analysis. The single amino acid substitutions at positions 124 and 135, although impairing the transformation functions, did not detectably alter the formation of the higher-apparent molecular-weight forms of the E1A products. PMID- 3025596 TI - Amino-terminal fragments of delta 1-pyrroline-5-carboxylate dehydrogenase direct beta-galactosidase to the mitochondrial matrix in Saccharomyces cerevisiae. AB - delta 1-Pyrroline-5-carboxylate (P5C) dehydrogenase, the second enzyme in the proline utilization (Put) pathway of Saccharomyces cerevisiae and the product of the PUT2 gene, was localized to the matrix compartment by a mitochondrial fractionation procedure. This result was confirmed by demonstrating that the enzyme had limited activity toward an externally added substrate that could not penetrate the inner mitochondrial membrane (latency). To learn more about the nature of the import of this enzyme, three gene fusions were constructed that carried 5'-regulatory sequences through codons 14, 124, or 366 of the PUT2 gene ligated to the lacZ gene of Escherichia coli. When these fusions were introduced into S. cerevisiae either on multicopy plasmids or stably integrated into the genome, proline-inducible beta-galactosidase was made. The shortest gene fusion, PUT2-lacZ14, caused the production of a high level of beta-galactosidase that was found exclusively in the cytoplasm. The PUT2-lacZ124 and PUT2-lacZ366 fusions made lower levels of beta-galactosidases that were mitochondrially localized. Mitochondrial fractionation and protease-protection experiments showed that the PUT2-lacZ124 hybrid protein was located exclusively in the matrix, while the PUT2 lacZ366 hybrid was found in the matrix as well as the inner membrane. Thus, the amino-terminal 124 amino acids of P5C dehydrogenase carries sufficient information to target and deliver beta-galactosidase to the matrix compartment. The expression of the longer hybrids had deleterious effects on cell growth; PUT2 lacZ366-containing strains failed to grow on proline as the sole source of nitrogen. In the presence of the longest hybrid beta-galactosidase, the wild-type P5C dehydrogenase was still properly localized in the matrix compartment, but its activity was reduced. The nature of the effects of these hybrid proteins on cell growth is discussed. PMID- 3025597 TI - Bidirectional promoter elements of simian virus 40 are required for efficient replication of the viral DNA. AB - Mutants of simian virus 40 (SV40) lacking parts of the 72- and 21-base-pair repeat regions were made deficient in large T antigen by recombination with dlA 4000, a mutant containing a frameshift deletion near the amino terminus of the T antigen genes. These double mutants were transfected into COS cells, and the amounts of replicated viral DNA were measured at various times thereafter. It was found that deletion of either the 72- or 21-base-pair repeat region did not significantly reduce the accumulation of viral DNA. However, cells transfected with mutants lacking both of these promoter elements accumulated 100-fold less viral DNA than cells transfected with wild-type SV40. This indicates that the 72- and 21-base-pair repeat regions are each sufficient for supplying a function required for efficient replication of SV40 DNA. In addition, the ability of either of these regions to support efficient replication was gradually reduced as the number of promoter elements within each was decreased. Since the 72- and 21 base-pair repeat regions bidirectionally induce transcription, our results indicate that bidirectional promoter elements play a role in the replication of viral DNA. However, fewer of these elements are required for efficient replication than for efficient transcription. PMID- 3025598 TI - Overproduction of protein p53 contributes to simian virus 40-mediated transformation. AB - The possible involvement of p53 overproduction in simian virus 40 (SV40)mediated transformation was studied by using the rat embryo fibroblast focus formation assay. Transformation by wild-type SV40 was enhanced two- to threefold by cotransfection of a plasmid overexpressing mouse p53. More significantly, such a plasmid could partially complement a transformation-defective deletion mutant of SV40. Hence, the ability of SV40 T antigen to induce high p53 levels may indeed be directly relevant to the viral transforming potential. PMID- 3025599 TI - A transcriptional enhancer and an interferon-responsive sequence in major histocompatibility complex class I genes. AB - The major histocompatibility complex class I antigens play an indispensable role in cell-cell interactions. Perturbation of their expression has been shown to have deleterious physiological consequences, including the escape of transformed cells from immune detection. In an attempt to understand how class I genes are regulated, we dissected the Ld gene to identify potential control regions. By using a test vector containing the simian virus 40 early promoter placed upstream of the bacterial chloramphenicol acetyltransferase (cat) gene, we demonstrated the presence of a transcriptional enhancer within the 5'-flanking region. The sequence is functional in both orientations and has been mapped within 350 base pairs upstream of the Ld transcriptional start site. Although human adenovirus 12 can suppress endogenous class I genes, it cannot down-regulate the activity of the transiently transfected cat gene which has been placed under the control of the Ld enhancer and promoter. Our results suggested that if the human adenovirus 12-induced function regulates the expression of class I genes by a trans mechanism, then its target site must not be within 1.9 kilobases of the 5' flanking region. Treatment of cells with interferon increases the accumulation of class I transcripts. Expression of the cat gene under the control of the Ld enhancer and promoter also can be up-regulated by interferon. Our study shows that the target sequence required for this enhancement resides, at least in part, within the same 350-base pair segment which contains the transcriptional enhancer. PMID- 3025600 TI - Excision repair functions in Saccharomyces cerevisiae recognize and repair methylation of adenine by the Escherichia coli dam gene. AB - Unlike the DNA of higher eucaryotes, the DNA of Saccharomyces cerevisiae (bakers' yeast) is not methylated. Introduction of the Escherichia coli dam gene into yeast cells results in methylation of the N-6 position of adenine. The UV excision repair system of yeast cells specifically responds to the methylation, suggesting that it is capable of recognizing modifications which do not lead to major helix distortion. The UV repair functions examined in this report are involved in the incision step of pyrimidine dimer repair. These observations may have relevance to the rearrangements and recombination events observed when yeast or higher eucaryotic cells are transformed or transfected with DNA grown in E. coli. PMID- 3025601 TI - Saccharomyces cerevisiae SPT3 gene is required for transposition and transpositional recombination of chromosomal Ty elements. AB - Mutations in the Saccharomyces cerevisiae SPT3 gene have dramatic effects on the expression of Ty elements and genes adjacent to the element. The SPT3 gene is essential for Ty transposition, because transposition of chromosomal Ty elements ceased when the SPT3 gene was replaced with the frameshift mutation spt3-101. Presumably, the elimination of transposition was due to the effect of the SPT3 gene product on Ty transcription; the transcripts of chromosomal Ty elements were largely abolished in the spt3-101 strain (F. Winston, K. J. Durbin, and G. R. Fink, Cell 39:675-682, 1984). Ty transcription in an spt3-101 strain could be reestablished by introduction of the pGTyH3 plasmid, in which transcription of the Ty element TyH3 is under the control of the GAL1 promoter; these plasmid derived Ty transcripts were SPT3-independent. Ty transposition resumed after galactose induction in spt3-101 strains containing the pGTyH3 plasmid. In spt3 mutants nearly all of the resulting transposition events derived from pGTyH3 plasmids and not from chromosomal elements. PMID- 3025603 TI - Gene-sized macronuclear DNA molecules are clustered in micronuclear chromosomes of the ciliate Oxytricha nova. AB - Following the sexual phase of its life cycle, the hypotrichous ciliate Oxytricha nova transforms a copy of its chromosomal micronucleus into a macronucleus containing short, linear DNA molecules with an average size of 2.2 kilobase pairs. In addition, more than 90% of the DNA sequences in the micronuclear genome are eliminated during this process. We have examined the organization of macronuclear DNA molecules in the micronuclear chromosomes. Macronuclear DNA molecules were found to be clustered and separated by less than 550 base pairs in two cloned segments of micronuclear DNA. Recombinant clones of two macronuclear DNA molecules that are adjacent in the micronucleus were also isolated and examined by DNA sequencing. The two macronuclear DNA molecules were found to be separated by only 90 base pairs in the micronuclear genome. PMID- 3025602 TI - Activation of the adenovirus and BK virus late promoters: effects of the BK virus enhancer and trans-acting viral early proteins. AB - We have examined the activation of the adenovirus major late promoter (MLP) by the cis-acting enhancer element of the human polyomavirus BK and by the trans acting simian virus 40 (SV40) T antigen and adenovirus E1A proteins. By using chloramphenicol acetyltransferase expression vectors, we found that the MLP (pLP CAT) was trans-activated in human and monkey kidney cells expressing the SV40 T antigen. In addition, the MLP could be cis-activated by the BK virus enhancer in both human and monkey kidney cells; approximately 20 times more chloramphenicol acetyltransferase was produced from expression vectors containing a hybrid promoter (BL), in which the BK enhancer was upstream of the MLP, than from expression vectors containing the MLP alone. This same level of enhancement of the MLP by the BK enhancer was observed in cells expressing the T antigen of SV40. However, in the 293 cell line, greater enhancement of MLP activity (70 fold) was observed with the BK enhancer sequence. In contrast, MLP activity in the 293 cell line was unchanged by the SV40 enhancer. In cotransfection experiments, MLP activity, augmented by the BK enhancer, could be further stimulated with a plasmid coding for the E1A gene products. By creating deletion mutants, we determined that the high-level activation of the hybrid BL transcriptional unit by the E1A proteins requires both MLP sequences and an intact BK virus enhancer. On the other hand, activation of the BL transcriptional unit by the T antigen did not require an intact enhancer sequence. Our results suggest that the SV40 T antigen and E1A proteins trans-activate the BL promoter by different mechanisms. We also demonstrate in cotransfection experiments that the BK late promoter is activated 45-fold by the SV40 T antigen. PMID- 3025604 TI - Comparison of the trans-activation capabilities of the human T-cell leukemia virus type I and II chi proteins. AB - The mechanism of cellular transformation by the human T-cell leukemia viruses (HTLVs) is thought to involve a novel retrovirus gene known as chi. The chi gene is essential for HTLV replication and acts by enhancing transcription from the viral long terminal repeat. By using the HTLV type I and II chi gene-coding regions inserted into a highly efficient expression vector, we directly compared the efficiencies of the two chi proteins to trans activate the HTLV type I and II long terminal repeats. We demonstrate that the two chi proteins have different patterns of trans activation. The patterns were highly reproducible in all mammalian cells tested. A different pattern of activation was observed in avian cells. These results suggest that the mechanism of trans activation involves specific cellular factors that are highly conserved throughout mammalian species but different in avian cells. Understanding the mechanism of trans activation by the chi gene product may provide insights into mechanisms of cellular transformation by HTLV. PMID- 3025606 TI - Transcriptional control signals of a herpes simplex virus type 1 late (gamma 2) gene lie within bases -34 to +124 relative to the 5' terminus of the mRNA. AB - The cis-acting DNA sequences required for regulated expression of a herpes simplex virus type 1 (HSV-1) late (gamma 2) gene were studied by using viruses containing specific deletions in the 5' transcribed noncoding and upstream regions of the HSV-1 glycoprotein C (gC) gene, a model gamma 2 gene. Nine mutant viruses which had variable 5' and 3' deletions within bases -569 to +124 relative to the 5' terminus of the gC mRNA were isolated. The mutants were isolated by a simple in situ hybridization screening procedure not requiring any prior selective pressure for or against expression of the gC gene. Analysis of RNA extracted from cells infected with individual mutants showed that the DNA sequences required for regulated expression of this gamma 2 gene lay within bases -34 to +124. This 158-base-pair fragment was sufficient to confer accurate and quantitative expression of gC mRNA and to maintain the stringent requirement on viral DNA replication for expression of this gene. Moreover, it was found that sequences located between -34 and +14 contained signals essential for expression of gC. To determine whether the -34 to +124 sequences would function as a gamma 2 promoter when moved to another region of the HSV-1 genome, the 158-base-pair fragment was substituted for the normal thymidine kinase promoter-regulatory sequences in the thymidine-kinase gene locus. Transcription of this chimeric gene was regulated as a gamma 2 gene in that its expression in infected cells was dependent on viral DNA synthesis. The only recognizable consensus sequence upstream of the transcription initiation site for this gene was the TATAAA sequence at -30. PMID- 3025607 TI - Minimal transcriptional enhancer of simian virus 40 is a 74-base-pair sequence that has interacting domains. AB - We have assayed the ability of segments of the simian virus 40 (SV40) 72-base pair (bp) repeat enhancer region to activate gene expression under the control of the SV40 early promoter and to compete for trans-acting enhancer-binding factors of limited availability in vivo in monkey CV-1 or human HeLa cells. The bacterial chloramphenicol acetyltransferase and the herpes simplex virus type 1 thymidine kinase genes were used as reporters in these assays. A 94-bp sequence located between SV40 nucleotides 179 and 272, including one copy of the 72-bp repeat, has been termed the minimal enhancer in previous studies. In the present study, we found that the 20-bp origin-proximal region located between nucleotides 179 and 198 was dispensable, since its removal caused only a slight reduction in enhancer activity. However, the deletion of another 4 bp up to nucleotide 202 abolished the enhancer activity. We propose that the minimal enhancer is a 74-bp sequence located between nucleotides 199 and 272, including 52 bp of one copy of the 72-bp repeat and a 22-bp adjacent sequence up to the PvuII site at 272. The nonamer 5' AAGT/CATGCA-3', which we term the K core, occurred as a tandem duplication around the SphI site at nucleotide 200, and we found that this duplication was essential for enhancement and factor-binding activities. A heterologous core element (which we term the C core), 5'-GTGGA/TA/TA/TG-3', identified earlier (G. Khoury and P. Gruss, Cell 33:313-314, 1983; Weiher et al., Science 219:626-631, 1983) also occurred in duplicate, with one of the copies located within the 22-bp sequence near nucleotide 272 present outside the 72-bp repeat. We provide direct evidence that this 22-bp sequence augments enhancer activity considerably. We also found that in addition to the heterologous interaction occurring normally between the K and C cores within the minimal enhancer, certain homologous interactions were also permitted provided there was proper spacing between the elements. PMID- 3025605 TI - Human homologs of TU transposon sequences: polypurine/polypyrimidine sequence elements that can alter DNA conformation in vitro and in vivo. AB - We previously have shown that homologs of the outer domain segment of the inverted repeat termini (IVR-OD) of the sea urchin TU transposons are conserved among multiple eucaryotic species, including humans. We report here that two cloned human DNA IVR-OD homologs, Hut2 and Hut17, consist of a series of tandem repeats of the trimer AGG/TCC, forming segments (313 and 221 base pairs in length, respectively) of polypurine/polypyrimidine (pPu/pPy or "Puppy") asymmetry in the two DNA strands; these are punctuated at certain sites with variant trimers, which are different for the two clones. Sequences homologous to the Hut2 pPu/pPy tract exist at multiple sites in the DNA of a wide variety of eucaryotes. Hybridization of human DNA with a Hut2 probe or with a previously described chicken DNA pPu/pPy sequence indicates that pPu/pPy sequences can be grouped into families distinguishable by the extent of their homology with each probe at different hybridization stringencies. Moreover, particular pPu/pPy tracts show species-specific differences in their distribution. Both the Hut2 and Hut17 pPu/pPy tracts are cleaved by S1 nuclease when tested on supercoiled plasmids. Most if not all of the 313-base-pair Hut2 pPu/pPy tract is also sensitive to S1 in its native location in HeLa cell chromatin, indicating that the sequence contains conformational information that can be expressed in vivo. This view is supported by evidence that exogenously derived Hut2 pPu/pPy tracts introduced into mouse L cells and integrated in chromatin can assume an S1-sensitive conformation. PMID- 3025608 TI - Structure of the protein encoded by the chicken proto-oncogene c-myb. AB - The retroviral oncogene v-myb arose by transduction of the chicken proto-oncogene c-myb. We isolated and sequenced cDNA that represents the entire coding domain of chicken c-myb. By transcribing the cDNA into mRNA in vitro and then translating the RNA, we were able to document the integrity of the cDNA and to identify the codon responsible for initiation of translation from c-myb. Two different alleles of v-myb are extant, one in the genome of avian myeloblastosis virus (AMV) and the other in the genome of erythroblastosis virus 26 (E26V). The proteins encoded by the AMV and E26V alleles of v-myb differ from the product of c-myb in three ways: at their amino termini, they lack 71 and 80 amino acids respectively; at their carboxy termini, they are deficient in 199 and 278 residues; and 11 substitutions of amino acids are scattered throughout the product of AMV allele, whereas the product of the E26V allele contains only a single substitution. The structural origins of tumorigenicity by v-myb and the biological functions of c myb remain enigmatic. The findings and molecular clones described here should now permit a systematic exploration of these enigmas. PMID- 3025609 TI - Crystallin gene expression and lentoid body formation in quail embryo neuroretina cultures transformed by the oncogenic retrovirus Mill Hill 2 or Rous sarcoma virus. AB - The lens-specific proteins alpha and delta crystallins and lentoid bodies, structures that follow a differentiation pathway similar to that of the lens, regularly appear after 4 to 5 weeks in quail embryo neuroretina monolayer cultures. We have investigated the effects of the avian oncogenic retroviruses Mill Hill 2 and Rous sarcoma virus on this process. Quail embryo neuroretina cells transformed by Mill Hill 2 virus were established into permanent cultures that synthesized alpha and delta crystallins and contained stem cells for the production of lentoid bodies. In contrast, transformation with the Rous sarcoma virus mutant tsNY-68 blocked the appearance of mRNA crystallins, but cytoplasmic alpha and delta crystallin mRNA and alpha crystallin appeared 44 h after a shift to the nonpermissive temperature. However, delta crystallins and lentoid bodies were only present after 7 days. The crystallins of transformed quail neuroretina cultures were immunologically indistinguishable from those of quail lenses and of normal quail embryo neuroretina cultures. PMID- 3025610 TI - Two functional alpha-tubulin genes of the yeast Saccharomyces cerevisiae encode divergent proteins. AB - Two alpha-tubulin genes from the budding yeast Saccharomyces cerevisiae were identified and cloned by cross-species DNA homology. Nucleotide sequencing studies revealed that the two genes, named TUB1 and TUB3, encoded gene products of 447 and 445 amino acids, respectively, that are highly homologous to alpha tubulins from other species. Comparison of the sequences of the two genes revealed a 19% divergence between the nucleotide sequences and a 10% divergence between the amino acid sequences. Each gene had a single intervening sequence, located at an identical position in codon 9. Cell fractionation studies showed that both gene products were present in yeast microtubules. These two genes, along with the TUB2 beta-tubulin gene, probably encode the entire complement of tubulin in budding yeast cells. PMID- 3025611 TI - Characterization of two nonallelic pairs of late histone H2A and H2B genes of the sea urchin: differential regulation in the embryo and tissue-specific expression in the adult. AB - Two nonallelic pairs of late H2A and H2B genes of the sea urchin Psammechinus miliaris were isolated on two different cosmid clones. The genes of cosmid PmL1 are separated by 11 kilobases of DNA and code for the late H2A-2 and H2B-2 variants. The genes of clone PmL2 are divergently transcribed with 1,060 base pairs of intergenic spacer DNA and code for novel variants of the H2A-2 and H2B-2 type. A comparison of the promoter sequences revealed little homology upstream of the TATA box with the exception of a 24-base-pair-long conserved sequence which is present at the same position in both late H2B promoters and part of which is identical with the "H2B-specific" 5' element. The mRNAs of the H2A and H2B genes of cosmid PmL1 reach their maximal levels early in the mesenchyme blastula embryo, whereas the transcripts of both genes of clone PmL2 accumulate maximally only later in the pluteus larva. In the adult sea urchin all four mRNAs are present in the tube foot but not in the intestine and lantern muscle. This pattern of differential expression in the embryo and tissue-specific expression in the adult suggests cell lineage-specific regulation of the late H2A-2 and H2B 2 genes. Another class of late histone genes represented by the H2A-3 and H2B-1 genes was shown to be expressed in all three adult tissues tested, whereas transcripts of the late H2A-1 genes could not be detected, suggesting that these genes are active exclusively during sea urchin development. PMID- 3025612 TI - Glycolytic gene expression in Saccharomyces cerevisiae: nucleotide sequence of GCR1, null mutants, and evidence for expression. AB - In Saccharomyces cerevisiae, the gcr mutation is known to have a profound effect on the levels of most glycolytic enzymes, reducing them to 5% of normal or less in growth on noncarbohydrates. Here I report the preparation of chromosomal gcr insertion and deletion mutations. The null mutations were recessive, were not lethal, and caused a pattern of glycolytic enzyme deficiency similar to that seen earlier for the gcr1-1 allele, including the partial inducibility by glucose of the residual enzyme activities. DNA sequence analysis showed that GCR1 encoded a protein of molecular weight 94,414, with a very low codon bias index, characteristic of several S. cerevisiae regulatory genes; adjacent 5' and 3' sequences contained elements suggesting that it was transcribed, polyadenylated, and translated. RNA gel transfer hybridization experiments with purified polyadenylated RNA and a probe complementary to the 5' portion of the open reading frame showed that Ger was expressed as a polyadenylated transcript. Together with previous work, the present results suggest that the Gcr product may be a transcriptional factor necessary specifically for the high-level transcription of a limited set of genes whose products, the enzymes of glycolysis, constitute a substantial fraction of cell proteins and are responsible for the primary metabolic flux in many cells. PMID- 3025613 TI - Initiation of simian virus 40 DNA replication in vitro: aphidicolin causes accumulation of early-replicating intermediates and allows determination of the initial direction of DNA synthesis. AB - Aphidicolin, a specific inhibitor of DNA polymerase alpha, provided a novel method for distinguishing between initiation of DNA synthesis at the simian virus 40 (SV40) origin of replication (ori) and continuation of replication beyond ori. In the presence of sufficient aphidicolin to inhibit total DNA synthesis by 50%, initiation of DNA replication in SV40 chromosomes or ori-containing plasmids continued in vitro, whereas DNA synthesis in the bulk of SV40 replicative intermediate DNA (RI) that had initiated replication in vivo was rapidly inhibited. This resulted in accumulation of early RI in which most nascent DNA was localized within a 600- to 700-base-pair region centered at ori. Accumulation of early RI was observed only under conditions that permitted initiation of SV40 ori-dependent, T-antigen-dependent DNA replication and only when aphidicolin was added to the in vitro system. Increasing aphidicolin concentrations revealed that DNA synthesis in the ori region was not completely resistant to aphidicolin but simply less sensitive than DNA synthesis at forks that were farther away. Since DNA synthesized in the presence of aphidicolin was concentrated in the 300 base pairs on the early gene side of ori, we conclude that the initial direction of DNA synthesis was the same as that of early mRNA synthesis, consistent with the model proposed by Hay and DePamphilis (Cell 28:767-779, 1982). The data were also consistent with initiation of the first DNA chains in ori by CV-1 cell DNA primase-DNA polymerase alpha. Synthesis of pppA/G(pN)6-8(pdN)21-23 chains on a single-stranded DNA template by a purified preparation of this enzyme was completely resistant to aphidicolin, and further incorporation of deoxynucleotide monophosphates was inhibited. Therefore, in the presence of aphidicolin, this enzyme could initiate RNA-primed DNA synthesis at ori first in the early gene direction and then in the late gene direction, but could not continue DNA synthesis for an extended distance. PMID- 3025614 TI - Mating type-like conversion promoted by the 2 micrograms circle site-specific recombinase: implications for the double-strand-gap repair model. AB - Double-strand breaks in DNA are known to promote recombination in Saccharomyces cerevisiae. Yeast mating type switching, which is a highly efficient gene conversion event, is apparently initiated by a site-specific double-strand break. The 2 micrograms circle site-specific recombinase, FLP, has been shown to make double-strand breaks in its substrate DNA. By using a hybrid 2 micrograms circle::Tn5 plasmid, a portion of which resembles, in its DNA organization, the active (MAT) and the silent (HML) yeast mating type loci, it is shown that FLP mediates a conversion event analogous to mating type switching. Whereas the FLP site-specific recombination is not dependent on the RAD52 gene product, the FLP induced conversion is abolished in a rad52 background. The FLP-promoted conversion in vivo can be faithfully reproduced by making a double-stranded gap in vitro in the vicinity of the FLP site and allowing the gap to be repaired in vivo. PMID- 3025615 TI - trans activation of an Epstein-Barr viral transcriptional enhancer by the Epstein Barr viral nuclear antigen 1. AB - Two regions of the Epstein-Barr virus (EBV) genome together make up an element, oriP, which acts in cis to support plasmid replication in cells that express the EBV nuclear antigen 1 (EBNA-1). The two components of oriP are a region containing a 65-base-pair (bp) dyad symmetry and a region containing 20 copies of a 30-bp direct repeat. Here we show that the 30-bp family of repeats of oriP can function as a transcriptional enhancer that is activated in trans by the EBNA-1 gene product. In either EBV-genome-positive cells or in cells that express EBNA 1, the 30-bp family of repeats, when positioned in either orientation upstream or downstream, enhances expression of the chloramphenicol acetyltransferase (CAT) gene expressed from either the simian virus 40 early promoter or the herpes simplex virus type 1 thymidine kinase promoter. The extent of transcriptional enhancement varies with the promoter and cell type. This enhanced CAT expression reflects an increased level of CAT mRNA and does not result from amplification of the plasmids expressing CAT. In addition, plasmids carrying the gene for resistance to hygromycin B and the 30-bp family of repeats yielded 10 to 100 times more hygromycin B-resistant colonies than the vector lacking the 30-bp family of repeats in both EBV-genome-positive cells and cells that express EBNA 1. EBNA-1 is known to bind to sequences within the 30-bp family of repeats (D. R. Rawlins, G. Milman, S. D. Hayward, and G. S. Hayward, Cell 42:859-868, 1985), and these trans- and cis-acting elements together have at least two functional roles: (i) they are required for DNA replication dependent upon oriP, and (ii) they can enhance expression of genes linked to the 30-bp family of repeats of oriP. PMID- 3025616 TI - Rat-brain Na,K-ATPase beta-chain gene: primary structure, tissue-specific expression, and amplification in ouabain-resistant HeLa C+ cells. AB - We deduced the complete amino acid sequence of the rat brain Na,K-ATPase beta subunit from cDNA. The rat brain beta-subunit exhibits a high degree of primary sequence and secondary structural homology with the human and Torpedo beta subunit polypeptides. Analysis of rat tissue RNA reveals that the beta-subunit gene encodes four separate mRNA species which are expressed in a tissue-specific fashion. In ouabain-resistant HeLa C+ cells, beta-subunit DNA sequences are amplified (approximately 20-fold) and beta-subunit mRNAs are overproduced relative to levels in parental HeLa cells. These results suggest that the beta subunit plays an important role in Na,K-ATPase structure-function and in the mechanism underlying cellular resistance to the cardiac glycosides. PMID- 3025617 TI - Structural and functional analysis of the MAL1 locus of Saccharomyces cerevisiae. AB - We describe the isolation of a 22.6-kilobase fragment of DNA containing the MAL1 locus of Saccharomyces cerevisiae. Our results demonstrate that the MAL1 locus, like the MAL6 locus, is a complex locus containing three genes. These genes were organized similarly to their MAL6 counterparts. We refer to them as MAL11, MAL12, and MAL13 and show that they are functionally homologous to the MAL61 (encoding maltose permease), MAL62 (encoding maltase), and MAL63 (encoding the positive regulator) genes of the MAL6 locus. Transcription from each of the three genes was analyzed in a strain carrying the undisrupted MAL1 locus and in strains carrying single disruptions in each of the MAL1 genes. The MAL1 and MAL1 loci were found to be highly sequence homologous and conserved throughout the region containing these three genes. The strain used to isolate the MAL1 locus also carried the tightly linked SUC1 gene. The SUC1 gene was found to be located on the same 22.6-kilobase fragment containing the MAL1 locus and 5 kilobases from the 3' end of the MAL12 gene. The meaning of these results with regard to the mechanism of regulation of maltose fermentation is discussed. PMID- 3025618 TI - Structural and functional analysis of chicken U4 small nuclear RNA genes. AB - Two distinct chicken U4 RNA genes have been cloned and characterized. They are closely linked within 465 base pairs of each other and have the same transcriptional orientation. The downstream U4 homology is a true gene, based on the criteria that it is colinear with chicken U4B RNA and is expressed when injected into Xenopus laevis oocytes. The upstream U4 homology, however, contains seven base substitutions relative to U4B RNA. This sequence may be a nonexpressed pseudogene, but the pattern of base substitutions suggests that it more probably encodes a variant yet functional U4 RNA product not yet characterized at the RNA level. In support of this, the two U4 genes have regions of homology with each other in their 5'-flanking DNA at two positions known to be essential for the efficient expression of vertebrate U1 and U2 small nuclear RNA genes. In the case of U1 and U2 RNA genes, the more distal region (located near position-200 with respect to the RNA cap site) is known to function as a transcriptional enhancer. Although this region is highly conserved in overall structure and sequence among U1 and U2 RNA genes, it is much less conserved in the chicken U4 RNA genes reported here. Interestingly, short sequence elements present in the -200 region of the U4 RNA genes are inverted (i.e., on the complementary strand) relative to their usual orientation upstream of U1 and U2 RNA genes. Thus, the -200 region of the U4 RNA genes may represent a natural evolutionary occurrence of an enhancer sequence inversion. PMID- 3025619 TI - New class of polyomavirus mutant that can persist as free copies in F9 embryonal carcinoma cells. AB - A small circular DNA was found extrachromosomally in a clone of F9 embryonal carcinoma (EC) cells at high copy numbers per cell. The DNA was cloned in plasmid pUC19. Restriction endonuclease analyses of the DNA indicated that the DNA (fPyF9) was a mutant of polyomavirus (Py) DNA and had a mutation in a noncoding regulatory region. There have been many reports on the isolation of Py mutants capable of replication in undifferentiated cells. However, fPyF9 was different from other Py mutants in the following aspects: it was harbored stably as a free copy at 1 X 10(4) to 5 X 10(4) copies per cell in EC cells; it replicated in undifferentiated cells better than in differentiated cells; it was extremely rearranged in the sequences of the enhancer B domain; and it carried in the enhancer B domain three copies of an exogenous sequence which does not exist in Py strain A2. From these observations, we propose a new class of Py EC mutant which has an autonomous state similar to that of plasmid and small circular DNA in host cells. PMID- 3025620 TI - Effects of poly[d(pGpT).d(pApC)] and poly[d(pCpG).d(pCpG)] repeats on homologous recombination in somatic cells. AB - Sequencing studies have shown that in somatic cells alternating runs of purines and pyrimidines are frequently associated with recombination crossover points. To test whether such sequences actually promote recombination, we have examined the effects of poly[d(pGpT).d(pApC)] and poly[d(pCpG).d(pCpG)] repeats on a homologous recombination event. The parental molecule used in this study, pSVLD, is capable of generating wild-type simian virus 40 DNA via recombination across two 751-base-pair regions of homology and has been described previously (Miller et al., Proc. Natl. Acad. Sci. USA 81:7534-7538, 1984). Single inserts of either a poly[d(pGpT).d(pApC)] repeat or a poly[d(pCpG).d(pCpG)] repeat were positioned adjacent to one region of homology in such a way that the recombination product, wild-type simian virus 40 DNA, could be formed only by recombination within the homologies and not by recombination across the alternating purine-pyrimidine repeats. We have found that upon transfection of test DNAs into simian cells, a poly[d(pCpG).d(pCpG)] repeat enhanced homologous recombination 10- to 15-fold, whereas a poly[d(pGpT).d(pApC)] repeat had less effect. These results are discussed in terms of the features of these repeats that might be responsible for promoting homologous recombination. PMID- 3025621 TI - Induction and repression of the urea amidolyase gene in Saccharomyces cerevisiae. AB - The DUR1,2 gene from Saccharomyces cerevisiae has been isolated on recombinant plasmids along with all DNA between the DUR1,2 and MET8 loci. DUR1,2 was found to encode a 5.7-kilobase transcript, which is consistent with our earlier suggestion that the DUR1 and DUR2 loci are two domains of a single multifunctional gene. Steady-state levels of the DUR1,2 transcript responded to induction and nitrogen catabolite repression in the same way as urea amidolyase activity. dal81 mutants (grown with inducer) contained barely detectable amounts of DUR1,2 RNA, whereas dal80 mutants (grown without inducer) contained the same amount as a wild-type induced culture. These observations support our earlier hypothesis that DUR1,2 is transcriptionally regulated, with control being mediated by the DAL80 and DAL81 gene products. We cloned the DUR1,2-Oh mutation and found it to be a Ty insertion near sequences required for complementation of dur1,2 mutations. The ROAM phenotype of the DUR1,2-Oh mutation is sharply different from that of cis dominant, DUR80 mutations, which enhance DUR1,2 expression but do not affect the normal control pattern of the gene. There is evidence that DUR80 mutations may also be Ty insertions, which generate phenotypes that are different from those in DUR1,2-Oh mutations. PMID- 3025622 TI - Developmental regulation of Dictyostelium discoideum actin gene fusions carried on low-copy and high-copy transformation vectors. AB - The Dictyostelium discoideum genome contains an estimated 17 to 20 actin genes. We report the identification of a new member of this multigene family, actin 15, and its complete nucleotide sequence and transcription initiation sites. We constructed transformation vectors carrying either the actin 15 promoter fused to the neomycin phosphotransferase gene from transposon Tn903 or the actin 6 promoter fused to the neomycin phosphotransferase gene from Tn5. Cells transformed with the actin 15 vector carried less than five copies of vector DNA, while cells transformed with the actin 6 vector carried more than 200 copies. In both cases, the vector appeared to be integrated into the chromosome as a tandem array. Gene fusion RNAs transcribed from the actin 15 and actin 6 vectors were regulated like endogenous actin genes during D. discoideum development. DNA sequences required for temporal and cell type-specific regulation of these genes were contained within 2.8 kilobases of 5' noncoding DNA for actin 15 and 0.7 kilobases of 5' noncoding DNA for actin 6. PMID- 3025623 TI - Varied interactions between proviruses and adjacent host chromatin. AB - Retroviruses integrated at unique locations in the host genome can be expressed at different levels. We have analyzed the preintegration sites of three transcriptionally competent avian endogenous proviruses (evs) to determine whether the various levels of provirus expression correlate with their location in active or inactive regions of chromatin. Our results show that in three of four cell types, the chromatin conformation (as defined by relative nuclease sensitivity) of virus preintegration sites correlates with the level of expression of the resident provirus in ev+ cells: two inactive proviruses (ev-1 and ev-2) reside in nuclease-resistant chromatin domains and one active provirus (ev-3) resides in a nuclease-sensitive domain. Nuclear runoff transcription assays reveal that the preintegration sites of the active and inactive viruses are not transcribed. However, in erythrocytes of 15-day-old chicken embryos (15d RBCs), the structure and activity of the ev-3 provirus is independent of the conformation of its preintegration site. In this cell type, the ev-3 preintegration site is organized in a nuclease-resistant conformation, while the ev-3 provirus is in a nuclease-sensitive conformation and is transcribed. In addition, the nuclease sensitivity of host sequences adjacent to ev-3 is altered in ev-3+ 15d RBCs relative to that found in 15d RBCs that lack ev-3. These data suggest that the relationship between preintegration site structure and retrovirus expression is more complex than previously described. PMID- 3025624 TI - Tripartite sequences within and 3' to the sea urchin H2A histone gene display properties associated with a transcriptional termination process. AB - We have defined a DNA sequence that behaves as an RNA polymerase II termination signal by using the human HeLa cell transient expression system. Surprisingly, this sequence is tripartite, including part of the coding region of the sea urchin H2A histone gene together with two separate sequences in the 3' flanking region of the gene. We demonstrate that this signal functions both in its normal gene environment and also when placed within the human alpha-globin gene. However, we have failed to detect a discrete 3' terminus. Rather, our data indicate the presence of an extremely heterogeneous series of nonpolyadenylated RNAs. These heterogeneous nonpolyadenylated RNAs are stable when transcribed from the intact histone gene but are highly unstable within the human alpha-globin gene. This provides evidence for the role of poly(A) in the stability of mRNA. PMID- 3025625 TI - Adenovirus E1a proteins repress expression from polyomavirus early and late promoters. AB - We have examined the effects of the E1a products of adenovirus types 5 and 12 on the expression of polyomavirus early and late promoters. In cotransfection experiments in HeLa cells, plasmids expressing the E1a region of adenovirus type 5 or 12 repressed both the early and late promoters of polyomavirus, and deletion analysis indicates that the polyomavirus enhancers were the target of the E1a repression. With mutants lacking enhancer sequences, the polyomavirus early promoter but not the late promoter was trans-activated by E1a. Chimeric mutant plasmids with deletions in the regulatory region that contained either the A enhancer or the B enhancer were repressed to the same extent, indicating that E1a can repress both elements. Polyomavirus variant plasmids with rearrangements in the regulatory region conferring activity in embryonal carcinoma stem cells were repressed by E1a as was the wild type, suggesting that the repressor function is quite general. We discuss a model in which the influence of E1a on the transcriptional activity of a gene is the sum of positive and negative effects on promoter and enhancer elements and discuss possible mechanisms of negative regulation of enhancer function. PMID- 3025626 TI - Accessibility of the promoter sequence in the J-chain gene is regulated by chromatin changes during B-cell differentiation. AB - The gene for the immunoglobulin M (IgM)-polymerizing protein, the J chain, is activated when the mature B cell is triggered to secrete pentamer IgM. Activation of the gene was found to be associated with chromatin changes in a 240-base-pair region at the 5' end of the gene. Analyses of lymphoid lines showed that the 5' region was resistant to nuclease digestion at the immature B-cell stage; it became slightly more accessible in mature B cells and cells at an early stage in the IgM response and then displayed an open, hypersensitive structure in IgM secreting cells. In addition, analyses of normal, mitogen-stimulated lymphocytes showed that the open hypersensitive structure was coinducible with J-chain gene expression. These results suggest that the 5' chromatin changes precede transcription, making control sequences within the site accessible to regulatory factors. PMID- 3025627 TI - A nuclear gene of Saccharomyces cerevisiae needed for stable maintenance of plasmids. AB - We have isolated host mutants of Saccharomyces cerevisiae in which the 2 microns plasmid is poorly maintained. All the mutants tested constituted one complementation group, which was designated map1 (maintenance of plasmid). Minichromosomes carrying a chromosomal replication origin and a centromere were affected in the mutants. Two types of hybrid plasmids generated in vivo and in vitro appeared to compensate for the mutations and had DNA regions containing multiple ARS (autonomously replicating sequence) or a set of 2 microns inverted repeat sequences. These results suggested that poor maintenance of plasmids was due to low levels of replication, probably at the initiation of replication. PMID- 3025628 TI - Purification from a human hepatoma cell line of a basic fibroblast growth factor like molecule that stimulates capillary endothelial cell plasminogen activator production, DNA synthesis, and migration. AB - A 17,500-dalton protein which stimulates plasminogen activator production in cultured bovine capillary endothelial cells has been purified from a SK-Hep-1 human hepatoma cell lysate by using heparin affinity chromatography and fast protein-liquid ion exchange chromatography. The purified molecule stimulated plasminogen activator production in a dose-dependent manner between 0.01 and 1 ng/ml. It also stimulated collagenase synthesis, DNA synthesis, and motility in capillary endothelial cells in the same concentration range. This molecule was identified as a basic fibroblast growth factor-like molecule on the basis of its biological activity, its affinity for heparin-Sepharose, and its cross-reactivity with a polyclonal antibody raised against the human placental basic fibroblast growth factor. PMID- 3025629 TI - Different tissue-specific expression of the amylase gene Amy-1 in mice and rats. AB - We cloned the rat alpha-amylase gene Amy-1 and compared its structure and expression with its mouse counterpart. The results showed that the general organization of the transcriptionally active rat Amy-1 gene was similar to that of its mouse counterpart; i.e., the rat gene also contained two independent transcriptional promoters. The distance between the two promoters in the rat gene was, however, more than double (6 kilobases) that measured in the mouse gene (2.8 kilobases). In addition, the rat genome also contained an independent, orphonlike version of the weaker Amy-1 promoter, which was transcriptionally silent. In spite of the similar overall organization of the Amy-1 genes in mouse and rat cells, an interesting difference was observed in the expression of the weak promoter in these two closely related rodents. In rats this promoter was significantly active only in liver cells, while in mice it was utilized with similar efficiencies in parotid, liver, and pancrease cells. Moreover, the transcripts produced in rat liver had a very heterogeneous population of 5' ends, located between 180 and 220 nucleotides upstream of the two homologous start sites observed for this promoter in mouse liver, even though the sequences around this region were strongly conserved between the two species. PMID- 3025630 TI - T-antigen-DNA polymerase alpha complex implicated in simian virus 40 DNA replication. AB - We have combined in vitro DNA replication reactions and immunological techniques to analyze biochemical interactions between simian virus (SV40) large T antigen and components of the cellular replication apparatus. First, in vitro SV40 DNA replication was characterized with specific origin mutants. Next, monoclonal antibodies were used to demonstrate that a specific domain of T antigen formed a complex with cellular DNA polymerase alpha. Several antibodies were identified that coprecipitated T antigen and DNA polymerase alpha, while others were found to selectively prevent this interaction and concomitantly inhibit DNA replication. DNA polymerase alpha also bound efficiently to a T-antigen affinity column, confirming the immunoprecipitation results and providing a useful method for purification of the complete protein complex. Taken together, these results suggest that the T-antigen-polymerase association may be a key step in the initiation of SV40 DNA replication. PMID- 3025631 TI - Deregulation of the c-myc oncogene in virus-induced thymic lymphomas of AKR/J mice. AB - A high frequency (greater than or equal to 65%) of thymomas induced by mink cell focus-forming virus 69L1 in AKR/J mice contain proviral integrations which are clustered 0.7-kilobase upstream of the c-myc oncogene predominantly in the opposite transcriptional orientation. Blot hybridization experiments showed that on the average there was only a twofold elevation of steady-state c-myc RNA in the thymomas as compared with levels in normal AKR/J thymocytes. Such an increase would not appear to be sufficient as a mechanism of oncogene activation in this system. In contrast, S1 nuclease analysis of transcripts initiated from the two known c-myc promoters indicated a strong shift in promoter usage in virtually all thymomas tested. In normal thymus the ratio of transcripts initiated from the proximal promoter P1 to the distal promoter P2 was 0.2 to 0.3. In contrast, most of the thymomas tested (18 of 23) showed an average P1/P2 ratio of 1.2 regardless of whether or not proviral integrations could be detected within a 21-kilobase EcoRI fragment containing the three c-myc exons. We conclude that alterations in P1/P2 ratios are good indicators of c-myc deregulation in thymic lymphomas. PMID- 3025632 TI - Sequences involved in accurate and efficient transcription of human c-myc genes microinjected into frog oocytes. AB - By microinjecting a series of deletion mutant constructs into Xenopus laevis oocytes, transcriptional control regions, two promoters, of the human c-myc gene were defined. In the case of the first promoter, sequences between -60 and -37 relative to the transcription start site contained an element essential for promoter activity. In the case of the second promoter, sequences between -66 and 56 relative to the initiation site appeared to be involved in accurate and efficient transcription. In both cases, the region identified as the essential promoter element contained GGGCGG or GGCGGG,GC box-like sequences, suggesting that c-myc gene promoter activity may be controlled by transcription factor Sp1 binding in the microinjected oocytes. PMID- 3025633 TI - The mouse immunoglobulin heavy-chain gene enhancer contains sequences that inhibit transcription in vitro in HeLa cell extracts. AB - The effect of the cell-specific mouse immunoglobulin heavy-chain gene (IgH) enhancer on transcription from heterologous promoter elements was studied in vitro with a HeLa whole-cell extract. No stimulation of transcription could be seen under conditions in which an activation was observed with the simian virus 40 enhancer. We found, however, that a specific segment of the IgH enhancer region contains sequences which inhibit transcription in vitro. PMID- 3025634 TI - Active X chromosome DNA is unmethylated at eight CCGG sites clustered in a guanine-plus-cytosine-rich island at the 5' end of the gene for phosphoglycerate kinase. AB - The 5' control region and first exon for human X-chromosome-linked phosphoglycerate kinase is contained in a G + C-rich island. We measured methylation at all HpaII sites in this 5' region of leukocyte DNA. By use of a blotting procedure that allows analysis of small DNA fragments, we found that the HpaII sites are entirely methylated when from an inactive X chromosome and entirely unmethylated when from an active one. In contrast, methylation of HpaII sites in more downstream regions of the gene is essentially the same in active and inactive X chromosomes. PMID- 3025635 TI - Chromatin structure of a P-element-transduced hsp-28 gene in Drosophila melanogaster. AB - The Drosophila hsp-28 gene was heat inducible when transduced to novel chromosomal sites even when no direct selection for transduced gene expression was imposed. The pattern of DNase I-hypersensitive sites 5' to the wild type and transduced copy of hsp-28 was similar. In addition, DNase I-hypersensitive sites occurred within the P-element sequences flanking transduced loci. PMID- 3025636 TI - In vivo competition of delta-crystallin gene expression by DNA fragments containing a GC box. AB - Expression of the chicken delta-crystallin gene 1 injected into the nuclei of mouse cells is lens specific. Coinjection of GC box-containing DNA fragments from delta-crystallin, simian virus 40 early, and herpes simplex virus type 1 tk promoters effectively suppressed delta-crystallin expression in the lens, but coinjection with DNA fragments not containing the GC box did not. This suppression was likely due to the competition of an Sp1-like transcription factor(s) and indicates involvement of the apparently ubiquitous factor(s) in the tissue-specific expression of the delta-crystallin gene. PMID- 3025637 TI - Abelson virus abrogation of interleukin-3 dependence in a lymphoid cell line. AB - Among several tyrosine-protein kinases, only v-abl could abrogate interleukin 3 dependence of a lymphoblastoid cell line; v-src and v-fps proteins gave partial or no interleukin 3 independence, respectively. Lymphokine independence was achieved via a nonautocrine mechanism. Direct involvement of c-myc in this process was not evident. PMID- 3025638 TI - Extensive mutagenesis of the nuclear location signal of simian virus 40 large-T antigen. AB - Site-directed mutagenesis was used to change Lys-128 of the simian virus 40 large T nuclear location signal to Met, Ile, Arg, Gln, Asn, Leu, or His. Except for the large-T antigen of the Arg mutation, which was present in cytoplasmic and nuclear compartments, the resultant proteins were unable to enter the nucleus. By contrast, mutations at other sites within the signal were generally less severe in their effect. In some cases (Lys-128 to Gln, Asn, and His), the apparently cytoplasmic variants were able to support limited plasmid DNA replication, suggesting that low levels of large-T antigen undetectable by immunofluorescence were present in the nucleus. Such mutants did not support viral DNA replication. We conclude that there is a strong requirement for a basic residue at position 128 in the large-T nuclear location signal, with Lys the preferred residue. PMID- 3025639 TI - Adenovirus 2 peptide IX gene is expressed only on replicated DNA molecules. AB - Transcription of the adenovirus type 2 peptide IX (pIX) gene was examined in transient expression assays. When a nonreplicating plasmid DNA containing the pIX gene was introduced into HeLa cells by the DEAE-dextran method, no pIX gene transcript was detected. In contrast, efficient transcription was observed in the cells transfected with a replicating plasmid containing the pIX gene. Adenovirus early genes did not affect the level of transcription of the pIX gene on either a nonreplicating or a replicating plasmid. Inhibition of plasmid replication with cytosine arabinoside prevented transcription of the pIX gene. By quantitative analysis of the amount of the pIX gene and its transcript in transfected cells, it was concluded that active transcription of the pIX gene occurred only on DNA molecules replicated in the cell. PMID- 3025640 TI - An unusual adenine phosphoribosyltransferase pseudogene is syntenic with its functional gene and is flanked by highly polymorphic DNAs. AB - A mouse adenine phosphoribosyltransferase (aprt) pseudogene that had previously been recovered from a BALB/c sperm DNA library possessed several unusual features. Its nucleotide sequence, like that of other processed pseudogenes, was colinear with its corresponding mRNA, but it was truncated at its 3' end and lacked a poly(A) tail. The pseudogene was 82% homologous with corresponding regions of the functional gene and had incurred mutations that included transitions, transversions, deletions, and a point insertion. Even though the pseudogene was truncated within the protein-coding region of the corresponding functional gene, it was flanked at both ends by 13-base-pair direct repeats. Curiously, the direct repeats exhibited homology to APRT mRNA at the site of pseudogene divergence. The pseudogene appeared to be common to BALB/c and A/J mice, but it was contained on a 3-kilobase EcoRI fragment in the former strain and a 4.5-kilobase EcoRI fragment in the latter. The BALB/c and apparently the A/J pseudogene both mapped to chromosome 8, which also contains the functional aprt gene. The DNA sequences immediately surrounding the pseudogene in the two strains appeared to be similar, suggesting that the BALB/c and A/J pseudogenes are allelic. However, DNA sequences more distal to the pseudogene in the two strains appeared to vary. Thus, the EcoRI polymorphism was not due to simple loss of an EcoRI site, but was more complex. The pattern of flanking restriction sites was different for each of several enzymes, consistent with extensive DNA rearrangement. Double digests of BALB/c and A/J genomic DNAs revealed complex polymorphisms on both sides of the pseudogene. The results were consistent with insertion, deletion, or other rearrangement of DNA sequences that flank the pseudogene and suggest that this region of mouse chromosome 8 may be a region active for mutation or recombination. PMID- 3025642 TI - Activation of an enhancerless gene by chromosomal integration. AB - Expression of enhancerless (E-) and enhancer-containing (E+) genes that are chromosomally integrated was examined. An E- plasmid (pE-cat) containing a chloramphenicol acetyltransferase (cat) gene linked to the simian virus 40 (SV40) early promoter or its E+ counterpart plasmid (pE+-cat) containing the SV40 enhancer was cotransfected into thymidine kinase (TK)-deficient L cells with a cloned tk gene. A number of TK+ transformants were isolated, and expression of the cointegrated cat gene in these cell lines was quantitatively determined by the assay of CAT activity. The results indicated unexpectedly that the E- cat gene was as actively expressed as the E+ cat gene. Analysis of CAT mRNA by primer extension indicated that the E- cat gene, as well as the E+ cat gene, was transcribed from the "native" initiation site contained in the SV40 early promoter region. The active expression of the E- cat gene was maintained in secondary TK+ transformants that arose by transfection with genomic DNA from the primary transformant. These results suggest that expression of the integrated E- cat gene is activated by endogenous enhancer elements. PMID- 3025641 TI - Binding in vitro of multiple cellular proteins to immunoglobulin heavy-chain enhancer DNA. AB - Seven protein-binding sites on the immunoglobulin heavy-chain (IgH) enhancer element have been identified by exonuclease III protection and gel retardation assays. It appears that the seven sites bind a minimum of four separate proteins. Three of these proteins also bind to other enhancers or promoters, but one protein seems to recognize exclusively IgH enhancer sequences. A complex of four binding sites, recognized by different proteins, is located within one 80-base pair region of IgH enhancer DNA. Close juxtaposition of enhancer proteins may allow protein-protein interactions or be part of a mechanism for modulating enhancer protein activity. All IgH enhancer-binding proteins identified in this study were found in extracts from nonlymphoid as well as lymphoid cells. These data provide the first direct evidence that multiple proteins bind to enhancer elements and that while some of these proteins recognize common elements of many enhancers, others have more limited specificities. PMID- 3025643 TI - Random isolation of gene activator elements from the human genome. AB - Long-range-acting gene activator elements were randomly isolated from the human genome by functional selection. HeLa cells were transfected with an enhancer trap, a plasmid containing an enhancerless xanthine-guanosine phosphoribosyltransferase (gpt) gene transcribed from the simian virus 40 early promoter, and stably transformed GPT+ cells were selected. From several transformants, human DNA sequences flanking the enhancer trap were cloned. Two gene activators (GA1 and GA2) were found in the cloned human DNAs. GA1 and GA2 showed strong enhancer activity both in a stable transformation assay and in a transient expression assay. They had functional properties similar to those of other known enhancers: GA1 and GA2 activated the expression of a linked gene over distances of at least 5 kilobases both upstream and downstream in an orientation independent fashion. GA1 may be required for the initial establishment of gene activation but was not essential for the maintenance of active expression. GA1 and GA2 were active not only in HeLa cells but also in other types of human cells, such as neuroblastoma cells. This indicates a limited but relatively broad cell type specificity. The HeLa genome contains multiple copies of GA1, while GA2 exists once in the genome. PMID- 3025645 TI - Topoisomerase inhibitors can selectively interfere with different stages of simian virus 40 DNA replication. AB - I have found that antineoplastic drugs which are known to be inhibitors of mammalian DNA topoisomerases have pronounced and selective effects on simian virus 40 DNA replication. Ellipticine, 4'-(9-acridinylamino)methanesulfon-m aniside, and Adriamycin blocked decatenation of newly replicated simian virus 40 daughter chromosomes in vivo. The arrested decatenation intermediates produced by these drugs contained single-strand DNA breaks. Ellipticine in particular produced these catenated dimers rapidly and efficiently. Removal of the drug resulted in rapid reversal of the block and completion of decatenation. The demonstration that these drugs interfere with decatenation suggests that they may exert their cytotoxic and antineoplastic effects by preventing the separation of newly replicated cellular chromosomes. Camptothecin rapidly breaks replication forks in growing Cairns structures. It is likely that the target of camptothecin is the "swivel" topoisomerase required for DNA replication and that it is located at or very near the replication fork in vivo. Evidence is presented that many of the broken Cairns structures are in fact half-completed sister chromatid exchanges. One pathway for the resolution of these structures is completion of the sister chromatid exchange to produce a circular head-to-tail dimer. PMID- 3025644 TI - Expression of a human cytomegalovirus late gene is posttranscriptionally regulated by a 3'-end-processing event occurring exclusively late after infection. AB - A phenomenon of posttranscriptional regulation has been previously identified in cytomegalovirus-infected human fibroblast cells (Wathen and Stinski, J. Virol. 41:462, 1982). A region typifying this phenomenon has been located within the large unique component of the viral genome (map units 0.408 to 0.423). Even though this transcriptional unit was highly transcribed at early times after infection, mRNAs from this region were only detectable on the polyribosomes after viral DNA replication. Thus, this region is believed to code for a late gene. Single-strand-specific nuclease mapping experiments of viral transcripts established that the transcriptional initiation sites and the 5' ends of a downstream exon were identical at early and late times. However, the late transcripts differed from the early transcripts by the processing of the 3' end of the viral RNAs. This involved either the removal of a distinct region of the transcript by the selection of an upstream cleavage and polyadenylation site or the differential splicing of the RNA molecule. The upstream cleavage and polyadenylation site was identified by nuclease mapping analyses and DNA sequencing. The 3'-end processing of these transcripts is necessary for the detection of these viral RNAs within the cytoplasm of the infected cell. We propose that human cytomegalovirus either codes for a factor(s) that is involved in the 3'-end-processing event at late times after infection or stimulates the synthesis of a host cell factor(s) involved in this complex regulatory event. This level of regulation may have an influence on the types of cells that permit productive cytomegalovirus replication. PMID- 3025646 TI - DNA rearrangement causes a high rate of spontaneous mutation at the immunoglobulin heavy-chain locus of a mouse myeloma cell line. AB - The spontaneous mutation rate of immunoglobulin genes expressed in myeloma cells is well above that of other genes expressed in these or in other cell types. The nature of such mutations in one myeloma cell line, MPC11, was explored at the molecular level. Included in this study were MPC11 variants representing 24 independent and spontaneous mutations affecting immunoglobulin secretion. Of the mutants studied, 19 had ceased immunoglobulin heavy chain (IgH) production (nonproducers), and 5 produced from as little as 1/1,000 to as much as 1/10 the amount of immunoglobulin produced by MPC11 (low producers). Only one of the MPC11 mutants (a nonproducer) showed no evidence of DNA rearrangement in or near the expressed IgH gene. The formerly expressed gamma 2b gene had been deleted in 18 of the 19 nonproducers. All of the low producers had undergone DNA rearrangement in or near the expressed IgH gene, and three of them produced immunoglobulin of a new heavy chain class. The cause for reduced heavy-chain synthesis in the low producers is not yet known. However, in several of these mutants, the defect appeared to be posttranscriptional. In these cell lines, steady-state IgH mRNA levels were much lower than in the parent cell line, while the heavy-chain gene transcription rate remained unchanged. PMID- 3025647 TI - AKXD recombinant inbred strains: models for studying the molecular genetic basis of murine lymphomas. AB - We analyzed the lymphoma susceptibility of 13 AKXD recombinant inbred mouse strains derived from AKR/J, a highly lymphomatous strain, and DBA/2J, a weakly lymphomatous strain. Of the 13 strains used, 12 showed a high incidence of lymphoma development. However, the average age at onset of lymphoma varied considerably among the different AKXD strains, suggesting that they have segregated several loci that affect lymphoma susceptibility. A relatively unambiguous classification of lymphomas was made possible by using histopathology in addition to detailed molecular characterization of rearrangements in immunoglobulin heavy and kappa light genes and in T-cell receptor beta-chain genes. Among the 12 highly lymphomatous strains, only 2 were identified that, like the parental AKR/J strain, died primarily of T-cell lymphomas. Three strains died primarily of B-cell lymphomas, and one strain primarily of myeloid lymphomas. Six strains were susceptible to both T-cell and B-cell lymphomas. Thus, these strains have segregated genes that affect both lymphoma susceptibility and lymphoma type and should prove to be useful models for studying the molecular genetic basis of murine lymphomas. PMID- 3025648 TI - Multiple control elements in the TRP1 promoter of Saccharomyces cerevisiae. AB - The TRP1 promoter generates two groups of mRNAs, transcript I and transcript II. The difference in size between the largest and smallest mRNAs is about 200 base pairs. A series of one-sided and internal deletions were constructed in vitro throughout the TRP1 promoter, and the effect of each deletion on transcription was assessed by Northern blotting. We showed that 395 base pairs of the TRP1 promoter were sufficient for the normal transcription of all RNAs and that the promoter contained two control domains. The control domain for transcript I consisted of one positive element and one negative element, while the control domain for transcript II contained two positive elements. The negative element, mapped between -293 and -318, expression of transcript I. Two regions of transcript I. Two regions (-280 to -236 and -235 to -209) were required for accurate initiation of transcript I. Each region contained sequences homologous to known consensus sequences of the TATA box. PMID- 3025649 TI - Structure of the Saccharomyces cerevisiae HO gene and analysis of its upstream regulatory region. AB - The HO gene product of Saccharomyces cerevisiae is a site-specific endonuclease that initiates mating type interconversion. We have determined the nucleotide sequence of a 3,129-base-pair (bp) segment containing HO. The segment contains a single long open reading frame encoding a polypeptide of 586 amino acids, which has unusual (unbiased) codon usage and is preceded by 762 bp of upstream region. The predicted HO protein is basic (16% lysine and arginine) and is calculated to have a secondary structure that is 30% helical. The corresponding transcript is initiated approximately 50 nucleotides prior to the presumed initiation codon. Insertion of an Escherichia coli lacZ gene fragment into the putative HO coding segment inactivated HO and formed a hybrid HO-lacZ gene whose beta-galactosidase activity was regulated by the mating type locus in the same manner as HO (repressed by a 1-alpha 2). Upstream regions of 1,360 and 762 bp conferred strong repression; 436 bp led to partial constitutivity and 301 bp to full constitutivity. Thus, DNA sequences that confer repression of HO by a1-alpha 2 are at least 250 nucleotides upstream of the transcription start point and are within 436 nucleotides of the HO initiation codon. The progressive loss of repression suggests that both the -762 to -436 and the -436 to -301 intervals contain sites for regulation by a1-alpha 2. The HO gene contains two distinct regions that promote autonomous replication of plasmids in S. cerevisiae. These regions contain sequences that are homologous to the two conserved sequences that are associated with ARS activity. PMID- 3025650 TI - Nonhomologous recombination in mammalian cells: role for short sequence homologies in the joining reaction. AB - Although DNA breakage and reunion in nonhomologous recombination are poorly understood, previous work suggests that short sequence homologies may play a role in the end-joining step in mammalian cells. To study the mechanism of end joining in more detail, we inserted a polylinker into the simian virus 40 T-antigen intron, cleaved the polylinker with different pairs of restriction enzymes, and transfected the resulting linear molecules into monkey cells. Analysis of 199 independent junctional sequences from seven constructs with different mismatched ends indicates that single-stranded extensions are relatively stable in monkey cells and that the terminal few nucleotides are critical for cell-mediated end joining. Furthermore, these studies define three mechanisms for end joining: single-strand, template-directed, and postrepair ligations. The latter two mechanisms depend on homologous pairing of one to six complementary bases to position the junction. All three mechanisms operate with similar overall efficiencies. The relevance of this work to targeted integration in mammalian cells is discussed. PMID- 3025651 TI - Multiple protein-binding sites in the 5'-flanking region regulate c-fos expression. AB - We tested sequences flanking the mouse c-fos gene for the ability to form specific DNA-protein complexes with factors present in crude nuclear extracts prepared from mammalian cells. Three such complexes were detected. One complex formed in a region necessary for the induction of c-fos expression by serum growth factors. Two additional complexes formed at sequences that contribute to basal c-fos promoter activity in vivo. These complexes represent three novel sequence-specific DNA-binding activities which appear to participate in the regulation of c-fos transcription. PMID- 3025653 TI - Immortalization of rat embryo fibroblasts by mutant polyomavirus large T antigens deficient in DNA binding. AB - We have identified a putative DNA-binding domain in polyomavirus large T antigen. Mutations introduced into the gene between amino acids 290 and 310 resulted in proteins that no longer bound to the high-affinity binding sites on the polyomavirus genome, showed no detectable nonspecific DNA binding, and were not able to initiate DNA replication from the viral origin. These mutant T antigen genes were introduced into rat embryo fibroblasts together with the neomycin resistance gene to allow selection for growth in the presence of G418. All the mutations tested facilitated the establishment of these cells in long-term culture at an efficiency indistinguishable from that of the wild-type protein. PMID- 3025652 TI - Determination of the orientation of an integral membrane protein and sites of glycosylation by oligonucleotide-directed mutagenesis: influenza B virus NB glycoprotein lacks a cleavable signal sequence and has an extracellular NH2 terminal region. AB - The membrane orientation of the NB protein of influenza B virus, a small (Mr, approximately 18,000) glycoprotein with a single internal hydrophobic domain, was investigated by biochemical and genetic means. Cell fractionation and protein solubility studies indicate NB is an integral membrane protein, and NB has been shown to be a dimer under nonreducing conditions. Treatment of infected-cell surfaces with proteinase K and endoglycosidase F and immunoprecipitation with a site-specific antibody suggests that the 18-amino-acid NH2-terminal region of NB is exposed at the cell surface. Oligonucleotide-directed mutagenesis to eliminate each of the four potential sites of N-linked glycosylation and expression of the mutant NB proteins in eucaryotic cells suggest that the two sites adjacent to the NH2 terminus are glycosylated. This provides further evidence that NB, which lacks a cleavable NH2-terminal signal sequence, has an exposed NH2 terminus at the cell surface. PMID- 3025655 TI - A noncatalytic domain conserved among cytoplasmic protein-tyrosine kinases modifies the kinase function and transforming activity of Fujinami sarcoma virus P130gag-fps. AB - Proteins encoded by oncogenes such as v-fps/fes, v-src, v-yes, v-abl, and v-fgr are cytoplasmic protein tyrosine kinases which, unlike transmembrane receptors, are localized to the inside of the cell. These proteins possess two contiguous regions of sequence identity: a C-terminal catalytic domain of 260 residues with homology to other tyrosine-specific and serine-threonine-specific protein kinases, and a unique domain of approximately 100 residues which is located N terminal to the kinase region and is absent from kinases that span the plasma membrane. In-frame linker insertion mutations in Fujinami avian sarcoma virus which introduced dipeptide insertions into the most stringently conserved segment of this N-terminal domain in P130gag-fps impaired the ability of Fujinami avian sarcoma virus to transform rat-2 cells. The P130gag-fps proteins encoded by these transformation-defective mutants were deficient in protein-tyrosine kinase activity in rat cells. However v-fps polypeptides derived from the mutant Fujinami avian sarcoma virus genomes and expressed in Escherichia coli as trpE-v fps fusion proteins displayed essentially wild-type enzymatic activity, even though they contained the mutated sites. Deletion of the N-terminal domain from wild-type and mutant v-fps bacterial proteins had little effect on autophosphorylating activity. The conserved N-terminal domain of P130gag-fps is therefore not required for catalytic activity, but can profoundly influence the adjacent kinase region. The presence of this noncatalytic domain in all known cytoplasmic tyrosine kinases of higher and lower eucaryotes argues for an important biological function. The relative inactivity of the mutant proteins in rat-2 cells compared with bacteria suggests that the noncatalytic domain may direct specific interactions of the enzymatic region with cellular components that regulate or mediate tyrosine kinase function. PMID- 3025654 TI - Neoplastic transformation of rat 3Y1 cells by a transcriptionally activated human c-myc gene and stabilization of p53 cellular tumor antigen in the transformed cells. AB - Transformed foci were obtained in rat 3Y1 fibroblasts cotransfected with pRmyc 27 (transcriptionally activated c-myc) and pSV2neo DNA. RmycY cell lines (1 to 7) were established from these foci. RmycY cells were small and round and contained enlarged nucleoli in the nucleus. The myc gene was expressed in these cell lines at a much higher level than in 3Y1 cells and at a level similar to that in HL-60 cells. These cell lines formed colonies in soft-agar culture and tumors in syngeneic rats transplanted with RmycY cells. Expression of the gene and colony formation in soft-agar culture were analyzed in subclones from RmycY cell line 1. A correlation between myc gene expression and the ability to form colonies in soft-agar culture was observed in these cells. Antibody against p53 cellular tumor antigen was detected in some sera from tumor-bearing rats. p53 cellular tumor antigen stabilized and accumulated in RmycY cells to the same extent as in simian virus 40-transformed cells. The results suggest that elevated c-myc expression and an increased amount of p53 cause 3Y1 cells to become a more tumorigenic cell line. PMID- 3025656 TI - Sequence and expression of the chicken beta 5- and beta 4-tubulin genes define a pair of divergent beta-tubulins with complementary patterns of expression. AB - We have determined the nucleotide sequence of the chicken beta 5 (c beta 5) tubulin gene. The gene displayed the coding structure common to all previously studied vertebrate beta-tubulin genes and was divided into four exon sequences interrupted by three intervening sequences (located between codons 19 and 20, within codon 56, and within codon 93). Comparison of the predicted polypeptide sequence encoded by c beta 5 with those of four other available chicken beta tubulin sequences revealed that c beta 5 encoded a highly divergent beta-tubulin polypeptide isotype which was distinguished from previously known sequences primarily by two discrete variable sequence domains. However, c beta 5 uniquely shared identity in 16 residue positions with another divergent chicken beta tubulin gene, c beta 4. These common sequences distinguished c beta 4 and c beta 5 from the remaining three chicken beta-tubulin genes. Analysis of the expression of c beta 5 and c beta 4 revealed a strikingly complementary pattern of gene expression: c beta 5 was expressed in a wide variety of cell and tissue types but not in neurons, whereas c beta 4 expression was detected uniquely in neuronal cells. Overall, these findings suggest the existence of two divergent families of beta-tubulin sequences in the chicken and further raise the possibility that the complementary expression of the c beta 4 and c beta 5 genes may fulfill a requirement for the presence of a divergent beta-tubulin polypeptide isotype in all cell types. PMID- 3025657 TI - Isolation and characterization of PRT1, a gene required for the initiation of protein biosynthesis in Saccharomyces cerevisiae. AB - We isolated a cloned DNA fragment containing PRT1, a gene required for the initiation of protein biosynthesis in Saccharomyces cerevisiae, by complementation of the temperature-sensitive prtl-1 mutation. The entire PRT1 gene is contained within a 3.2-kilobase-pair segment of the cloned DNA in YEp13 H1.2. Southern blot analysis demonstrated that PRT1 is a single copy gene which is transcribed into a 2.3-kilobase RNA. We determined the direction of transcription and mapped the 5' and 3' ends of the gene. PMID- 3025658 TI - Modulation of novel-length DOPA decarboxylase transcripts by 20-OH-ecdysone in a Drosophila melanogaster Kc cell subline. AB - The induction of DOPA decarboxylase (DDC) activity by 20-OH-ecdysone (20-OHE) in a subline of Drosophila melanogaster Kc cells was investigated. Cells cultured in the continuous presence of the steroid hormone exhibited a 96-h temporal lag prior to a peak of DDC enzyme activity while arrested in the G2 phase of the cell cycle. The concentration of Ddc RNA increased sixfold between 72 and 96 h after initial exposure to hormone. Similarly, this increase was correlated temporally with a 26-fold increase in DDC enzyme activity. The Kc Ddc primary transcript, processing intermediate, and mature mRNA all were approximately 500 nucleotides longer than the corresponding transcripts observed for newly eclosed adult D. melanogaster. In vitro translation of poly(A)+ RNA from Kc cells resulted in an immunoprecipitable polypeptide which exhibited similar mobility on sodium dodecyl sulfate gels to that of DDC synthesized in vitro by larval epidermal poly(A)+ RNA. PMID- 3025659 TI - Multiple hormone-inducible enhancers as mediators of differential transcription. AB - Sets of genes under a common regulatory control in a given cell type are often differentially transcribed. The possibility that this differential transcription can be modulated by the number or strength of cis-acting regulatory sequences associated with a given gene was tested by using the glucocorticoid-responsive enhancer element associated with the mouse mammary tumor virus promoter. Results indicate that differential levels of hormone-inducible gene expression can be modulated in an additive way by the number of glucocorticoid-responsive enhancers associated with this promoter. Realization of these effects shows little preference for position of the additional elements with respect to the promoter. When sequences that bind the glucocorticoid receptor in vitro with somewhat lower affinity than the enhancer were tested, these additive effects were not detected. The results support that differential transcription of genes subject to a common regulatory control can be mediated, at least in part, by the number or strength of their associated cis-acting regulatory sequences. PMID- 3025660 TI - The MES-1 murine enhancer element is closely associated with the heterogeneous 5' ends of two divergent transcription units. AB - The location in the mouse genome of the 149-base pair MES-1 element, previously isolated by its ability to restore expression to an enhancerless selectable gene, was analyzed. The active moiety of the single-copy MES-1 element is located between the 5' ends of two divergent transcription units, SURF-1 and SURF-2, both of which specify more than one mRNA species by differential splicing. The heterogeneous 5' ends of the SURF transcripts are separated by only 50 to 75 base pairs, and this sequence possesses a high G + C content (65%) and contains neither the TATA and CAAT box motifs normally associated with many highly expressed genes nor the GC box motif (Sp1-binding site) associated with a number of housekeeping genes. Although MES-1 appears to have enhancerlike properties when linked to heterologous genes, its normal genomic location suggests that it functions as a bidirectional promoter. Thus, MES-1 may represent a new class of enhancer-promoter element. PMID- 3025661 TI - Functional analysis of the role of the A + T-rich region and upstream flanking sequences in simian virus 40 DNA replication. AB - One boundary of the minimal origin of replication of simian virus 40 DNA lies within the A + T-rich region. Deletion of only a few bases into the adenine thymine (AT) stretch results in a DNA template which is defective for replication both in vivo and in vitro (B. Stillman, R. D. Gerard, R. A. Guggenheimer, and Y. Gluzman, EMBO J. 4:2933-2939, 1985). In the present study, such deletion mutations have been reconstructed into a simian virus 40 genome containing an intact early promoter-enhancer region. The resulting mutants synthesized wild type levels of T antigen, but were defective for replication and would not form plaques on CV-1 monkey cells. Replication-competent phenotypic revertants were selected after transfection of large quantities of the replication-defective viral DNAs into CV-1 cells. DNA sequence analysis showed that most of these revertants contained insertions or point mutations which partially regenerate the length of the AT stretch. These genotypic alterations were shown to be responsible for the revertant phenotype by replication analysis in vivo of subcloned revertant origin fragments. In general, our results emphasize the importance of the AT region to simian virus 40 origin function. However, one revertant retained the altered AT region but deleted six nucleotides upstream. Experiments using this mutant indicate that the 21-base-pair repeats identified as part of the early transcriptional promoter may compensate for defects in simian virus 40 DNA replication in vivo caused by mutations in the A + T-rich region when positioned at an appropriate distance from the core origin. PMID- 3025662 TI - The adenine-thymine domain of the simian virus 40 core origin directs DNA bending and coordinately regulates DNA replication. AB - The simian virus 40 origin of replication contains a 20-base-pair adenine-thymine rich segment with the sequence 5'-TGCATAAATAAAAAAAATTA-3'. The continuous tract of eight adenines is highly conserved among polyomaviruses. We used single-base substitutions to map structural and functional features of this DNA. Mutations in the AAA and AAAAAAAATT sequences significantly reduce DNA replication and thus identify two sequence-specific functional domains or a single domain with two parts. The AAAAAAAATT sequence also determines a DNA conformation that is characteristic of DNA bending. Single-base mutations in this domain change the degree of net bending, presumably by altering the length and location of the bending sequence. Thus, DNA bending in the correct conformation and location may be a structural signal for replication in polyomavirus origins and perhaps in other origins of replication with consecutive runs of adenines. The first five base pairs (TGCAT) of the 20-base-pair segment and the T between the AAA and AAAAAAAATT domains serve a sequence-independent function that may establish proper spacing within the core origin. PMID- 3025664 TI - Molecular basis for heterogeneity of the human p53 protein. AB - The human p53 tumor antigen comprises several physically distinct proteins. Two p53 proteins, separable by polyacrylamide gel electrophoresis, are expressed by the human transformed cell line SV-80. The individual cDNAs which code for these proteins were isolated and constructed into the SP6 transcription vector. The proteins encoded by these clones were identified by in vitro transcription with the SP6 vector and translation in a cell-free system. p53-H-1 and p53-H-19 cDNA clones code for the faster- and slower-migrating p53 protein species, respectively, of SV-80. The in vitro-expressed proteins of p53-H-1 and p53-H-19 had the same antigenic determinants and were structurally indistinguishable from their in vivo counterparts. By expressing defined restricted cDNA fragments in vitro, the region of heterogeneity between the respective cDNAs was located at the 5' end of the cDNAs. Exchanging the 5' fragments of interest and expressing the chimeric clones in vitro confirmed that the DNA heterogeneity was responsible for the difference in the electrophoretic mobility of these proteins. The sequences of the two cDNAs revealed a single base pair difference (G versus C) in the coding region of the clones. This sequence difference resulted in an arginine being coded for in clone p53-H-1 and a proline being coded for at the equivalent position in clone p53-H-19. This variation accounted for the change in the electrophoretic mobility of the individual p53 protein species. PMID- 3025663 TI - The chicken ubiquitin gene contains a heat shock promoter and expresses an unstable mRNA in heat-shocked cells. AB - A chicken genomic library was screened to obtain genomic clones for ubiquitin genes. Two genes that differ in their genomic location and organization were identified. One gene, designated Ub I, contains four copies of the protein-coding sequence arranged in tandem, while the second gene, Ub II, contains three. The origin of the two major mRNAs that are induced after heat shock in chicken embryo fibroblasts was determined by generating DNA probes from the 5'-and 3'-noncoding regions of the two genes. Both mRNAs are transcribed from Ub I, the larger being the unspliced precursor of the smaller. A 674-base-pair intron was located within the 5'-noncoding region of Ub I. The second gene, Ub II, does not appear to code for an RNA species in normal or heat-shocked chicken embryo fibroblasts. The expression of ubiquitin mRNA during heat shock and recovery was examined. Addition of actinomycin D before heat shock completely abolished the response of ubiquitin mRNA to the stress. Analysis of the stability of the mRNA during recovery revealed that the mRNA accumulated during the heat shock is rapidly degraded with a half-life of approximately 1.5 h, suggesting a specialized but transient role for ubiquitin during heat shock. PMID- 3025665 TI - A single Saccharomyces cerevisiae upstream activation site (UAS1) has two distinct regions essential for its activity. AB - Several site-directed mutagenesis regimens were used to generate single- and multiple-base substitutions in the upstream activation site UAS1 of the Saccharomyces cerevisiae CYC1 gene. Mutations resulting in large reductions in activity of the site lie in two distinct regions. Six single-base changes in a region A, between -288 and -285, all resulted in a 15-fold reduction in activity. Synthetic sites built up solely of multimers of the -289 to -285 sequence ACCGA behaved as carbon catabolite-sensitive UASs. In addition, substitution mutations in a second region, at nucleotides -266 and -265, virtually eliminated UAS1 activity. These mutations abolished the binding of a heme-dependent protein factor in vitro. Thus, UAS1 contains two essential regions both of which are required for its activity. PMID- 3025666 TI - Transcriptional control of the mouse prealbumin (transthyretin) gene: both promoter sequences and a distinct enhancer are cell specific. AB - The mouse genomic clone for the prealbumin (transthyretin) gene was cloned, and its upstream regulatory regions were analyzed. The 200 nucleotides 5' to the cap site when placed within a recombinant plasmid were sufficient to direct transient expression in HepG2 (human hepatoma) cells, but this DNA region did not support expression in HeLa cells. The sequence of the 200-nucleotide region is highly conserved between mouse and human DNA and can be considered a cell-specific promoter. Deletions of this promoter region identified a crucial element for cell specific expression between 151 and 110 nucleotides 5' to the RNA start site. A region situated at about 1.6 to 2.15 kilobases upstream of the RNA start site was found to stimulate expression 10-fold in HepG2 cells but not in HeLa cells. This far upstream element was invertible and increased expression from the beta-globin promoter in HepG2 cells. Unlike the simian virus 40 enhancer, the prealbumin enhancer would not stimulate beta-globin synthesis in HeLa cells, and even the simian virus 40 enhancer did not stimulate the prealbumin promoter in HeLa cells. Thus, we identified in the prealbumin gene two DNA elements that respond in a cell-specific manner: a proximal promoter including a crucial sequence between 108 and -151 nucleotides and a distant enhancer element located between 1.6 and 2.15 kilobases upstream. PMID- 3025667 TI - Activation of oncogenicity of the c-rel proto-oncogene. AB - Reticuloendotheliosis virus strain T (Rev-T) induces a lethal lymphoma in young birds and transforms avian lymphoid cells in vitro. The transforming gene of Rev T, v-rel, was derived from the turkey proto-oncogene c-rel. Comparison of the nucleotide sequences of v-rel and c-rel indicates that in addition to several internal amino acid changes relative to c-rel, p59v-rel has amino acid sequences at both ends derived from the reticuloendotheliosis virus strain A-related virus env gene (K. C. Wilhelmsen, K. Eggleton, and H. M. Temin, J. Virol. 52:172-182, 1984). In this report, the v-rel sequences important for transformation were defined by constructing recombinant retroviruses in which c-rel sequences replaced the analogous v-rel sequences. These recombinant viruses expressing chimeric proteins were tested for their ability to transform spleen cells in vitro and to induce tumors in young chickens. Activation of the oncogenicity of c rel in Rev-T required alteration of the amino terminus and the central region of the protein. Deletion of the noncoding sequences 3' to c-rel and of most of the helper virus-related env sequences was necessary for the formation of Rev-T. PMID- 3025668 TI - In vitro cleavage of the simian virus 40 early polyadenylation site adjacent to a required downstream TG sequence. AB - Exogenous RNA containing the simian virus 40 early polyadenylation site was efficiently and accurately polyadenylated in in vitro nuclear extracts. Correct cleavage required ATP. In the absence of ATP, nonpoly(A)+ products accumulated which were 18 to 20 nucleotides longer than the RNA generated by correct cleavage; the longer RNA terminated adjacent to the downstream TG element required for polyadenylation. In the presence of ATP analogs, alternate cleavage was not observed; instead, correct cleavage without poly(A) addition occurred. ATP-independent cleavage of simian virus 40 early RNA had many of the same properties as correct cleavage including requirements for an intact AAUAAA element, a proximal 3' terminus, and extract small nuclear ribonucleoproteins. This similarity in reaction parameters suggested that ATP-independent cleavage is an activity of the normal polyadenylation machinery. The ATP-independent cleavage product, however, did not behave as an intermediate in polyadenylation. The alternate RNA did not preferentially chase into correctly cleaved material upon readdition of ATP; instead, poly(A) was added to the 3' terminus of the cleaved RNA during a chase. Purified ATP-independent cleavage RNA, however, was a substrate for correct cleavage when reintroduced into the nuclear extract. Thus, alternate cleavage of polyadenylation sites adjacent to a required downstream sequence element is directed by the polyadenylation machinery in the absence of ATP. PMID- 3025669 TI - [Type III osteogenesis imperfecta associated with hypophosphatemic vitamin D resistant rickets]. AB - In a 7 year old girl presenting with bone deformities, dwarfism, and a history of recurrent fractures osteogenesis imperfecta had been diagnosed at birth. Although she had been hospitalized several times, radiologic signs of rickets remained unnoticed. Laboratory data proved existence of hypophosphatemic vitamin D resistant type of rickets, which was effectively treated with 1 alpha hydroxycholecalciferol and phosphorus substitution. The combination of osteogenesis imperfecta type III and hypophosphatemic rickets may be coincident. It proves, however, the necessity to consider the possible simultaneous occurrence of two rare diseases. The therapeutic consequences could be important. PMID- 3025670 TI - Prenatal ACTH and corticosteroids, development and behavior in rat. PMID- 3025671 TI - Neuropeptide-neurotransmitter interaction due to GRF and CRF stimulation in experimental and clinical conditions during development. PMID- 3025672 TI - Adrenergic influences on sexual differentiation of the rat brain. PMID- 3025673 TI - Neuroendocrine response to oestrogen in transsexual men. PMID- 3025674 TI - [Cloning in Escherichia coli of the mutant gene coding the diphtheria toxin]. AB - Bacteriophage beta 45 of Corynebacterium diphtheriae was harvested. The extracted DNA of the bacteriophage was digested by the restriction endonuclease BamHI and inserted into the BamHI cleavage site of pUC19 vector plasmid. Plasmid pNVY5 containing a mutant gene crm45 of diphtheriae toxin in a 3.9 bpn fragment was isolated from the hybrid plasmids obtained. Cell free extracts of E. coli strain TG1 (pVNY5) contain the nontoxic protein crm45 possessing the specific enzymatic activity of diphtheriae toxin (ADP ribosylation on wheat elongation factor two). According to orientation of BamHI fragment in pNVY5 plasmid it is concluded that the crm45 gene is expressed using its own promoter. PMID- 3025675 TI - [Conjugative R-plasmid of facultative methylotrophic bacteria Pseudomonas sp. M]. AB - 50 Md conjugative plasmid, designated pM3, has been found in the cells from natural isolates of Pseudomonas sp M. The plasmid determines the resistance to tetracycline and streptomycin and is capable of conjugative transfer between the cells of Pseudomonas and Escherichia coli. The conjugative derivatives of pM3 deleted for 14 Md of molecular mass were isolated after acridine dyes treatment of cells harbouring plasmid pM3. The discovered plasmid was not shown to belong to IncP1 incompatibility group. PMID- 3025676 TI - [Different effect of mutations in Escherichia coli K12 dna-genes on the transposition of Tn5- and Tn10-elements]. AB - The effect of mutations in dnaA(dnaA46), dnaG(dnaG3), dnaC (dnaC1 and dnaC2) and dnaB genes on transposition of two transposons, Tn5 and Tn10, from bacteriophage lambda genome into the chromosome of host cells has been studied. Transposition was performed at permissive temperatures for the mutant recipients. The mutations in dnaA, dnaC, dnaG genes were shown to decrease the transposition of Tn10 for some orders of magnitude as compared with transposition registered in wild type cells. Independence of Tn5 transposition of the above mentioned genes was demonstrated, providing evidence on the different modes of transposition of these two Tn-elements. PMID- 3025677 TI - [Isolation of Yersinia pestis plasmids with transposon markers]. AB - Transposons Tn1, Tn7, Tn9, Tn10 have been inserted into each of three known plasmids in Yersinia pestis and have been shown to mutagenize the different plasmid genes. The marked plasmids are shown to be transduced by bacteriophage P1 cml clr 100 ts in intrageneric crosses. The genes of 61-65 Md plasmid were found to be impaired with high frequencies by Tn9 and Tn10 insertions blocking the synthesis of fraction I antigen. The genes are also impaired in course of transduction of transposon marked plasmids. PMID- 3025679 TI - [Deletion of the phage lambda att80 Tn9 genome; the type of intramolecular recombination]. AB - The intramolecular deletion-generating recombination which transforms lambda bacteriophage genomes into the plasmids (named pLS) proved to be site-specific to a certain extent. Using electron microscopy heteroduplex analysis three preferential sites for this recombination were found in seven independent pLS isolates studied. Att-sites were not registered to be involved in the formation of deletions in isolates studied. It was shown that recombination operating in our system was independent of the phage int and bacterial recA genes. PMID- 3025678 TI - [Transcription, in frog oocytes and transgenic mice, of recombinant plasmids with long terminal repeats of retrovirus proviruses]. AB - RNA-DNA hybridization was used to study the transcription efficiency of the genes controlled by the long terminal repeats (LTR) from two different retroviral proviruses (exogenous provirus of the chicken Rous sarcoma virus and endogenous xenotropic provirus of the mouse). The oocyte nuclei of Xenopus laevis and the liver of transgenic mice were used as the transcription systems. The transcription efficiency of genes with the above mentioned promoters was shown to be roughly the same for both systems. Thus, the LTR of two proviruses of different origin possess the promoters of comparable strength and do not show any marked tissue or species specificity. PMID- 3025680 TI - [Identification of the plasmid pLG13 coding for the DNA modification-restriction system EcoRV and properties of strains carrying this plasmid]. PMID- 3025681 TI - [Cloning of DNA fragments of the genetic transfer factor pAP39]. AB - Large HindIII digested fragments of the plasmid pAP39 have been cloned on the cosmid vector pHC79. The study of the structure of HindIII fragments of plasmid pAP39 in the recombinant plasmids has shown that these fragments are represented by f1 + f2 fragments from the plasmid pAP1055, by f1 + f6 fragments from the plasmid pAP1056, by f2 + f3 fragments from the plasmid pAP1057 and by two f3 fragment from the plasmid pAP1058. Physical maps of the recombinant plasmids have been constructed. The plasmid pAP39 is shown to contain two functionally active tra regions. PMID- 3025682 TI - [Effect of mutation changes in RNA-polymerase and transcription termination factor rho on expression of various operons in E. coli]. AB - Six mutations, impairing DNA polymerase of E. coli in combination with the wild type gene for rho factor or ts-mutation rho 15 have been studied in relation to the expression of seven operons having different types of regulation. The expression of genes for glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase is shown to be constitutive and resistant to mutationally altered RNA polymerase and rho factor. The expression of genes for adenine phosphoribosyltransferase and of deo operon is regulated by rho dependent attenuators with attenuation being lifted incomplete medium. Mutation rho 15 decreases the level of enzymes of thr and lac operons independent of mRNA levels of these operons. Mutation rho 15 effect on posttranscriptional level is modified by mutations damaging RNA polymerase. The data obtained suppose RNA polymerase to affect all stages of realization of genetic information, beginning with promoter recognition and RNA synthesis and including the protein synthesis on mRNA. PMID- 3025683 TI - [Hybrid plasmid pBS251 containing genes for n-alkane degradation]. AB - The strain of Pseudomonas aeruginosa BS316 utilizing H-alkanes of the C6-C12 series (Alk+) harbours the 96 Md plasmid pBS250. The use of plasmid RP4 to mobilize Alk+ markers in conjugal transfer to Pseudomonas aeruginosa and Pseudomonas putida has resulted in isolation of transconjugants resistant to antibiotics (due to genes coded by plasmid RP4) and capable of growth on H alkanes. A transconjugant from this series harbours plasmid pBS251, a derivative of plasmid RP4 containing the genes for octane and octanol catabolism. A fragment of DNA inserted into RP4 has a mol mass 3.8 Md, possesses two restriction sites for EcoRI, one site for PstI, is not restricted by SmaI and BamHI restriction endonucleases, and is localized in the region 4.5-5.7 Md on the physical map of plasmid RP4. PMID- 3025684 TI - [Identification of the plasmid R1drd-19 region complementing the mutant phenotype recB- in Escherichia coli K12 strain]. AB - Plasmid R1-19 and its copy number mutants markedly increase the recombinational efficiency of a recB- strain of E. coli K12 and its resistance to the lethal action of UV and mitomycin C. These effects are associated with the appearance of a new ATP-dependent exonuclease activity in recB- cells known to be deficient in the ATP-dependent exonuclease V. Using hybrid plasmids carrying different EcoRI fragments of R1-19 (in the pSF124 vector), the gene(s) responsible for effect of R1-19 in recB-cells were localized in the EcoRI-C fragment (8.5 MD) belonging to the RTF portion of R1-19. Expression of the gene(s) in hybrid plasmids depends on the orientation of EcoRI-C fragment in the vector. The copy number of the EcoRI-C fragment was not strictly correlated with the degree of expression of the effects in the recB- mutant. PMID- 3025685 TI - [Recombinant plasmids capable of integration into the chromosome of the cyanobacterium Anacystis nidulans R2]. AB - Recombinant plasmids, series pIAB and pIAH, have been constructed by insertion of BamHI or HindIII chromosomal fragments from Anacystis nidulans R2 into the tet gene of plasmid pACYC184. Plasmids pIAB and pIAH are stably maintained in Escherichia coli cells and transfer the CmR marker in transformation of Anacystis nidulans. Blot hybridization technique has shown the formation of CmR clones in transformation to result from integration of plasmid pACYC184 with the chromosome of cyanobacterium. PMID- 3025687 TI - [Molecular biology of natural and constructed in vitro recombinants of human adenoviruses and SV40]. AB - This review is devoted to the molecular and biological properties of natural and artificial Ad-SV40 hybrids. The conditions of adenovirus and SV40 recombinant formations are discussed. The principal one is the ability of the C-terminal portion of the SV40 T-antigen to carry a helper function for growing human adenoviruses in the cultures of simian cells, in which the adenoviruses themselves are not reproduced. The data about the structural and functional organization of the hybrid RNA and proteins, tumor properties of defective and nondefective Ad-SV40 recombinants are provided. A special chapter is devoted to the construction of Ad-SV40 hybrids in vitro. The possibility of using them not only for the investigation of adenoviral properties, but for the super production of different proteins is marked. Alongside the corresponding region of SV40 genome may perform the function of a selection marker for the creation of eucaryotic vectors on the basis of adenoviruses. PMID- 3025686 TI - [Variants of the plasmid pAS8 delta with increased frequency of integration into Rhodopseudomonas sphaeroides chromosome--the result of IS8-element duplication]. AB - The possible participation of IS8 and IS elements of Rhodopseudomonas sphaeroides in cointegrate formation by chromosome of the purple bacterium and plasmid pAS8 121 delta has been studied. The plasmid derivatives having deleted Tn7 have been studied. Plasmid integration into the chromosome of the purple bacterium is shown to be mediated by IS8 element of the plasmid. Plasmid derivatives having the integration potential increased for two orders were isolated by a series of intergeneric conjugational crosses during which plasmid pAS8-121 delta was transferred from Rhodopseudomonas sphaeroides (cointegrate of plasmid and chromosome) to Escherichia coli (plasmid in an autonomous state) and back to Rhodopseudomonas sphaeroides. The restriction analysis of plasmid DNA digested by Hpal and Smal restriction endonucleases has revealed the tandem duplications of IS8 in plasmids capable of integration into the chromosome of the purple bacterium with a high frequency. PMID- 3025688 TI - [Antiviral activity of proteinase inhibitors in cultured cells infected with alpha-viruses]. AB - The ability of synthetic inhibitors of trypsin-like (TLCK) and chymotrypsin-like (TPCK) proteinases and natural antiproteinase oligopeptides of animal (aprotinin) and microbial (enzistatin) origin to suppress multicycle replication of different alpha viruses (Semliki, Sindbis, Venezuelan equine encephalomyelitis viruses) in cultured cells was studied. Antiviral activity was found to be induced by TPCK and aprotinin (Gordox). These compounds were shown to reduce virus yield 100-fold and to prevent the involvement of cells into infection process. The mechanisms of antiviral activity and chemotherapeutic possibilities of antiproteinase compounds are discussed. PMID- 3025689 TI - [Recovery of activity of the gene coding for tetracycline resistance in the plasmin pBRS188]. AB - The spontaneous recovery of activity of tet gene deleted of the promoter region was studied. Plasmid pBRS188 was used as a model for studying this problem. The plasmid has the fragment of tet gene of pBR322, from which it originates, between the sites of restriction endonucleases EcoRI and HindIII cleavage resulting in inactivation of tet promoter. E. coli cells harbouring the plasmid were shown to revert the TcR phenotype with the frequency 10(-9). The gene activation coincided with intraplasmid recombination revealed by restriction analysis. In some cases the recovery of tet gene activity coincided with the formation of multimeric plasmids. PMID- 3025691 TI - [Expression of cloned gene for methyltransferase from Bacillus subtilis bacteriophage SPbetaB]. AB - Expression of the methyltransferase gene from Bacillus subtilis lysogenizing phage SP beta B was studied by analyzing the sensitivity of the hybrid plasmid DNAs to restriction by the enzymes BspRI, Hpall and Mspl. The gene produces the methylase M. BsuP beta BI with specificity for 5'-GGCC. The fragment carrying the SP beta B derived gene also directs the synthesis in E. coli of a second methylase activity (M. BsuP beta BII) with 5'-CCGG specificity. Indirect evidence suggests that the two SP beta B modification activities are encoded by the same gene. PMID- 3025690 TI - [Effectiveness of incorporation of spermine-condensed viral DNA into liposomes. Interaction of liposomes with cells]. AB - DNA from bacteriophage lambda or monkey adenovirus type 7 have been condensed with spermine and included into three types of liposomes: ethereal, obtained by inverted phases technique and by Ca++ fusion. Compact DNA form presents a tor with 100 nm external diameter and 430 nm width. It can be included into 100 nm or larger liposomes. Total preparation contains 15-18% of the required liposomes. Liposomal fractions with included DNA were separated from empty liposomes by step gradient of ficoll 400. Liposomal fraction having included DNA contains 15-18% of common lipid. Liposomal interaction with monkey cell cultures has been studied. PMID- 3025692 TI - [Characteristics of mitochondrial DNA from Candida spec. EH15]. AB - The restriction endonucleases EcoRI, PstI and BglII were used to digest the mitochondrial DNA from Candida spec. EH 15. Molecular masses of single digested DNA fragments were calculated. The mitochondrial DNA size (28-30 x 10(6) D) comprises 2/3 of the mitochondrial DNA from Saccharomyces cerevisiae. PMID- 3025693 TI - [Characteristics of incomplete reproduction cycle of ts-mutant of influenza virus A at non-permissive temperature]. AB - Incomplete reproduction cycle of influenza virus A/134/17/57, the attenuation donor being used for preparation of recombinant vaccine strains, hs been analyzed with the use of molecular biology methods. Virus A/134/17/57 with two mutations in P3, NP and M genes remains capable of synthesis of viral polypeptides that are devoid of ability to be inserted into cellular plasma membrane, when the virus is propagated in MDCK culture at non-permissive temperature. The process is preceded by defects in the process of formation of RNP structures connected, evidently, with deficient synthesis of viral RNA due to mutations in the gene coding for P3 protein. PMID- 3025694 TI - [Restriction mapping of genetic transfer factor pAP39]. AB - A model of calculation of molecular weights of fragments EcoRI, Hind III and PvuI is formulated. A restriction site map of factor pAP39 is constructed automatically. Sites to EcoRI and PvuI are distributed in the segment with molecular weight 9.1 MD. PMID- 3025695 TI - [Mutagenic effect during transduction of (Gm-Km)R markers of the R323 plasmid in Yersinia pestis]. AB - Genes of resistance to some aminoglycoside antibiotics from plasmid R323 were transduced by bacteriophage P1 cml clr100 ts in Yersinia pestis. The resistance markers were capable of insertion into the chromosome or plasmid in the recipient cells causing the mutagenic effect. The results obtained suggest the transposon nature of plasmid fragment coding for gentamicin-kanamycin resistance. PMID- 3025697 TI - [Plasmid recombination stimulated by restriction endonuclease EcoRI in vivo: formation of recombinant plasmids in recA+-cells of E. coli]. AB - The possible participation of restriction endonuclease EcoRI in recombination of compatible nonhomologous plasmids in E. coli cells has been studied. To study the process, plasmids RP4 and R245 have been transferred by conjugation into the recipient cells of E. coli harbouring one of isogenic plasmids, pSA14 and pSA25, different for the genes coding restriction endonuclease EcoRI. The genetic analysis of transconjugant phenotypes, coded by the plasmids, has permitted to register the recombinant plasmids after compatibility of parent plasmids in E. coli cells. Recombination of plasmid RP4 with the plasmid pSA14, carrying EcoRI genes, has been registered in E. coli cells, producing the restriction endonuclease, while plasmid recombination has not been found in the cells harbouring plasmid pSA25, isogenic for all genes, except for EcoRI genes, with plasmid pSA14. Restriction endonuclease EcoRI is concluded to stimulate site specific recombination of nonhomologous compatible plasmids in vivo. EcoRI mediated recombination of plasmid R245 with plasmid pSA14 is discussed. PMID- 3025696 TI - [A system for insertion of heterologous DNA into chromosome of group H streptococci]. AB - A system for insertion of genes into the chromosome of group H streptococci has been elaborated. It consists of a recipient strain (Gallis GS10/1), having the fragments of lambda L-47-1 bacteriophage DNA inserted into the chromosome, and lambda 202 vector. The constructed system suggests the preliminary cloning of genes in E. coli cells with their subsequent insertion into the chromosome of group H streptococci. PMID- 3025698 TI - [Isolation of a restriction endonuclease with HindIII-specificity from Bordetella bronchiseptica]. AB - Site-specific restriction endonuclease BbrI has been found in bacteriophage resistant strain B. bronchioseptica 4994. The technique was elaborated for purification of BbrI to the stage free of nuclease and phosphatase contamination. The yield of purified enzyme is 6000-20 000 units per 10 g of biomass. BbrI recognises and cleaves the same DNA sequence as HindIII with the formation of four-nucleotide cohesive ends. The simplicity of cultivation, security for human, presence of the single restriction endonuclease and the high level of its production make B. bronchioseptica 4994 a promising producer of BbrI restriction endonuclease, isoshizomeric to HindIII, for use in experimental practice in industry. PMID- 3025699 TI - [Cloning of pectate-lyase genes of Erwinia chrysanthemi in Escherichia coli cells]. AB - Erwinia chrysanthemi DNA fragment digested by restriction endonuclease EcoRI and carrying the gene EC16 determining the synthesis of pectatelyase with Rf 0.20 and mol. mass 40kD has been cloned in plasmid pUC 9 plasmid in Escherichia coli HB101 cells. Three genes for pectatelyases of Erwinia chrysanthemi ENA49 have been cloned in vector phage lambda 47.1 in Escherichia coli cells. Two genes determining the synthesis of pectatelyases with Rf 0.06 and 0.19 and mol. masses 40 kD and 39 kD have been cloned as a part of an 7 kb Eco RI-fragment, that suggested their close location on the chromosome of Erwinia chrysanthemi ENA49. All of the cloned pectatelyase genes are expressed constitutively with pectatelyases accumulating in periplasm and being unable to secret into the cultural medium. PMID- 3025700 TI - [Molecular cloning of provirus sequences of Rauscher leukemia virus from mouse erythroleukemia cell genome]. AB - Provirus from a component of Rauscher leukaemia virus (RLV) has been cloned. The provirus (the size of 5000 b. p.) contains two LTR sequences and shares expressed sequence homology with Mo-MuLV. Restriction analysis and determination of the LTR nucleotide sequence and of the site from 3'-end of proviral genome have shown the cloned provirus to be the SFEV component of RLV. LTR from this cloned provirus contains all sites necessary for transcription: CAAT and TATA sequences, "cap" site and polyadenylation signal. The LTR of the cloned provirus from SFEV component of RLV has been shown to function as a promoter in E. coli cells. PMID- 3025701 TI - [Restriction map of pesticinogenicity plasmid pYP1 of Yersinia pestis]. AB - The restriction map of Yersinia pestis pesticinogenicity plasmid pYP1 has been constructed with the use of 18 restriction endonucleases. Plasmid dimensions (6.3 Md) have been specified, the genes for pesticin synthesis, for pesticin immunity protein, fibrinolysin and plasmocoagulase have been localized by molecular cloning of single plasmid DNA fragments in vector plasmid pBR322. PMID- 3025702 TI - [Effect of mutations in recB, recC, sbcB and recF genes controlling the homologous recombination in Escherichia coli cells on transposition of Tn1]. AB - The mutations in the genes controlling the homologous DNA recombination in Escherichia coli cells effect the efficiency of Tn1 transposition. Mutations in recB and recC genes decrease 50-fold the frequencies of Tn1 transposition. Introduction of an additional mutation in sbcB gene increase transposition frequency for three orders as compared with the one registered in wild type cells. Inactivation of sbcB gene in the wild type cells does not affect transposition significantly. Mutation in recF gene results in the great decrease of transposition when it is introduced into multiple recBC sbcB mutant, but not into the wild type bacteria. The possibility of two pathways for Tn1 transposition existing in Escherichia coli cells is discussed, as well as possibility of existence of similar stages in transposition and recombination controlled by the same genes. PMID- 3025703 TI - [Separation of modification methylation and restriction enzymes from Shigella sonnei 47]. AB - Two systems for DNA host specificity have been demonstrated for Shigella sonnei cells, SsoI and SsoII. The aim of the present work was to separate the modificating methylases and restriction endonucleases from Shigella sonnei and to study the modificating functions of methylases MSsoI and MSsoII. The possibilities to separate the methylation and restriction enzymes by column chromatography on affine, ionoexchange and hydrophobic sorbents were analyzed. The scheme for separation of methylases and restriction endonucleases of Shigella sonnei was elaborated, consisting of the fractioning of total preparation on phenyl-sepharose and subsequent isoelectrofocusing on ampholines. The modification functions of MSsoI and MSsoII methylases obtained by this technique and devoid of concomitant restriction endonucleases were studied. The in vitro experiments have shown the acceptor DNA methylated by MSsoI or MSsoII to be RSsoI or RSsoII restriction proof. PMID- 3025704 TI - [Viability and radioresistance of polB1 ssb-1 mutants of Escherichia coli K12]. PMID- 3025705 TI - [Translocation of ampicillin transposon Tn1 in Escherichia coli cells during sexual reproduction]. AB - The efficiency of Tn1 transposition has been shown to increase considerably in course of bacterial conjugation. Usually, the frequency of Tn1 transposition from plasmid pSA2001, a derivative of RP4, into the chromosome never exceeded 0.1% per cell. Percentage of His+ transconjugants, marked by transposon Tn1 during conjugation between Hfr donor, carrying plasmid pSA2001, and auxotrophic recipient, was about 30%. Transposon Tn1 transfer into the recipient cells does not depend on the recA+ gene function in donor cells or on conjugative transfer of plasmid pSA2001. The transfer requires the recA+ gene function in recipients as well as the Hfr function in donor cells. Southern's blot-hybridization revealed the insertion of transposon Tn1 into the different sites of the chromosome of His+ transconjugants. The transposon inserted during conjugation retains the ability to potential further translocation into new sites on the chromosomal DNA. PMID- 3025706 TI - [Characteristics of RecA-independent recombination of plasmids in E. coli cells producing restriction endonuclease EcoRI]. AB - The restriction endonuclease EcoRI dependent recombination of compatible plasmids has been studied in RecA cells of Escherichia coli. Plasmid RP4 and the isogenic ColE1 type plasmids pSA14 or pSA25, differing in restriction-modification RM EcoRI genes, have been used to study this type of recombination. EcoRI dependent recombination of plasmids is demonstrated in RecA cells and, thus, is independent of general system of homologous recombination. The classes of recombinant plasmids isolated from RecA cells differ from the classes isolated from wild type cells. Levels of tetracycline resistance conferred by plasmid RP4 are shown to be dependent on the alleles of RecA+ gene, being extremely low in RecA cells. This property is demonstrated to be useful for obtaining the multicopy recombinant plasmids resulting from EcoRI dependent recombination in RecA cells of Escherichia coli. PMID- 3025707 TI - [Properties of the plasmid pFT15/10-1 isolated from the vaccine strain of Francisella tularensis]. AB - Plasmid, designated pFT15/10-1, was isolated from Francisella tularensis vaccine strain 15/10. The plasmid is presented by the homogeneous 5.02 +/- 0.054 Md monomeric circular DNA molecules in electron microscopic preparations. Plasmid size is 7-7.3 kb as defined by electrophoresis in agarose gel. The restriction analysis has revealed that plasmid pFT15/10-1 possesses a single specific cleavage site for restriction endonuclease EcoRI, two sites for restriction endonucleases BamHI, BgIII, HincII, HindIII, PstI, three sites for BglI and SalI, some for AluI, TagI, MvaI, CfrI. Plasmid is not digested by restriction endonucleases SmaI, XmaI, KpnI, MluI. Restriction map of the plasmid was constructed for most frequently used restriction endonucleases. PMID- 3025708 TI - [Synthesis of aminoglycoside phosphotransferase is under control the PL-promoter of phage lambda]. AB - The recombinant plasmid pKP145 PL has been constructed containing the gene for aminoglycosidephosphotransferase (APT). Expression of the APT gene is under the control of lambda bacteriophage PL promoter. Escherichia coli cells harbouring this plasmid synthesize APT in quantity up to 13-15% of the total cellular protein. The technique for isolation of APT from superproducing cells has been elaborated. Preparations of the enzyme devoid of contaminating bacterial proteins have been obtained. PMID- 3025709 TI - [New specific endonucleases PaeI and PaeII from Pseudomonas aeruginosa]. AB - Specific endonuclease activities have been found it two Pseudomonas aeruginosa strains. Isolation and purification of enzymes and determining their specific activities have permitted one to find out that PaeI is an isoshizomer of SphI and digests the sequence 5'-GCATG C-3'. Another isolated enzyme PaeII is an isoshizomer of SmaI and cleaves DNA in a fragment 5'-CCC GGG-3'. The use of PaeI and PaeII enzymes in genetical engineering and their advantages are discussed. PMID- 3025710 TI - [Isolation of Escherichia coli K12 mutants with deficient in precise excision of transposons]. AB - Plasmid pNM1, the derivative of R100.1, has been constructed by insertion of transposon Tn5 into structural tet genet (Tn10) of the parental plasmid. The frequency of precise excision of Tn5 from plasmidic genome is 10(-5). The high frequency of precise excision obtained in this system permits one, to use it for isolation of mutants having low frequencies of precise excision. Two mutants were isolated in which the frequencies of precise excision of Tn5 were decreased for two orders. The pex1 and pex2 mutations responsible for the effect decrease the precise excision of Tn5 from R100.1 as well as from RP4 genomes. PMID- 3025711 TI - [Development of bacteriophage Mu in E. coli gyrBts mutant strain]. AB - The study deals with Mu growth in cells carrying a temperature-sensitive mutation in the gene of DNA gyrase B subunit. At a nonpermissive temperature the Mu growth is shown to be blocked in the host gyrB ts mutant both on infection and on prophage induction. Mu DNA does not get integrated in the host chromosome upon the infection of mutant cells, as demonstrated by DNA-DNA hybridization experiments. In the case of prophage induction in mutant cells, as opposed to the wild type cells early mRNA synthesis is practically fully inhibited while the total RNA synthesis is three times reduced after 20 min of induction. The transcription of phage DNA associated with the changed superhelicity of DNA in the cell. PMID- 3025712 TI - [Comparative study of non-conjugative R-plasmids from enterobacteria and Pseudomonas aeruginosa]. AB - Nonconjugative R-plasmids pBS76 and pBS94 (Sm Su), pBS95 and pBS96 (Sm Su Ap) isolated from clinical strains of Pseudomonas aeruginosa and plasmids pKMR281 pKMN284 (Sm Su), pKMR285-pKMR286 (Sm Su Tc) isolated from clinical strains of enterobacteria have been studied. Restriction maps of these plasmids are presented in the paper with some of plasmid genes for antibiotic resistance localized on them. The resistance determinants of plasmids pBS95 and pBS96 are shown to be included in transposon Tn3612 analogous to Tn3. Plasmids pBS76, pBS94 96 are of the wide host range and belong to incompatibility group P4 (IncQ). Plasmids pKMR281-pKMR286 are mutually incompatible and share the conspicuous DNA homology. They are inherited only by enterobacteria and are compatible with IncQ plasmids but in contrast to them are mobilized by RP4 plasmid with lower frequency. PMID- 3025713 TI - [Sedimentation characteristics of virion RNA of Machupo virus reproducing in the presence of actinomycin D]. AB - Actinomycin D treatment (0.005-05 g/ml) of Vero and BHK-21 cells infected with Machupo virus suppressed the synthesis of ribosomal RNAs but did not affect the production of infectious Machupo virus. Virion RNAs contained 3 high molecular weight RNA species: 28-31 S, 22-24 S and 18 S. In the presence of actinomycin D [3H]-uridine incorporated only in 30-31 S and 22-24 S RNA species. The data are supported by previous results which show that Machupo virus genome contains two RNA species: "large" (30-31 S) and "small" (22-24 S). PMID- 3025715 TI - [New yeast vectors containing the autonomously replicating sequences from Candida maltosa genome]. AB - Two different DNA sequences from the yeast Candida maltosa confer the ability to replicate autonomously to the yeast integrative vector pLD700 on which they are cloned. The recombinant plasmids pLD701 and pLD702 with autonomously replicating sequences (ARS) from Candida maltosa and LEU2 gene from Saccharomyces cerevisiae transform the auxotrophic strain S. cerevisiae DC5 with the efficiency 3-5 x 10(3) per microgram of DNA. Like other yeast vectors harbouring ARS, these plasmids are not stable in yeast cells. Restriction and hybridization analyses have revealed the pLD701 plasmid to contain ARS from chromosomal DNA of C. maltosa. Plasmid pLD701 appears to be a useful vector for yeast transformation. PMID- 3025714 TI - [The role of chromosome unwinding proteins in the regulation of DNA repair and replication]. AB - DNA repair and replication are regulated by cellular capability to support the definite optimal level of single strand DNA binding optimal level of single strand DNA binding proteins (SSB). SSB deficiency as well as it's overproduction in the cell cause, by means of different mechanisms, the destruction of DNA macromolecules due to impairment in timing of the nucleotides hydrolysis and resynthesis reactions. This asynchronization hinders the normal processing of DNA repair and replication. PMID- 3025716 TI - [Preparative isolation of myxovirus glycoproteins and the study of their immunogenic properties]. AB - Preparative isolation of glycoproteins from ortho- and paramyxoviruses is described. The purified concentrated virus has been treated with nonionic detergent MESK with subsequent removal of viral cores by centrifugation. Supernatant was sterilized by filtration through the nuclear filters and cleared from detergent by dialysis. Glycoproteins obtained have not contained contaminating cellular or core viral proteins or viral shell lipids. In the absence of detergent, glycoproteins have formed the peculiar mycelial complexes. Biological activity of glycoproteins was kept at high level. Glycoproteins output at isolation from different strains of influenza viruses A, B and Sendai virus varied from 75 to 98%. Immunogenetic study of the preparations obtained has demonstrated their capability to stimulate the formation of antibodies against both viral glycoproteins comparable with the capability of intact virus. The obtained level of immunity was enough to protect organism against homologous infection. Samples of glycoproteins obtained are up to standards for subunit vaccines, and the technique of their preparation is perspective as far as the production of vaccine preparations is concerned. PMID- 3025717 TI - [Effect of dioxidine, antineoplastic agents and other mutagens on the precise excision of Tn1 and Tn10 transposons in E. coli K12]. AB - Induction of precise excision of transposons Tn1 and Tn10 from the genes met::Tn1 and cys::Tn10 by chemical agents, having mutagenic and DNA damaging activities, has been studied. The drugs dioxydin, NMU, photrin, phopurine, thiophosphamid, rongeron as well as sodium azide, 2-NP, DDDTDP are shown to differ in their ability to stimulate the precise excision of transposons of different classes and in the efficiency of stimulated process. Results of the present paper are in proof of the possible using of experimental model, based on registering the precise excision of transposons, for screening the mutagenic and cancerogenic activities of chemical agents from the environment. PMID- 3025718 TI - [Plasmids of the cyanobacterium Synechocystis sp. 6803]. AB - Three cryptic plasmids have been isolated from cyanobacterium Synechocystis sp. 6803::pSS2 (1.4 Md), pSS3 (36 Md), pSS4 (60 Md). Plasmid DNA was isolated in Cs Cl-EB density gradient and analyzed by gel electrophoresis and electron microscopy by gel electrophoresis and electron microscopy techniques. The restriction map is constructed for plasmid pSS2 having the cleavage sites for Sau3a, HincII, HindIII, MspI restriction endonucleases. The plasmid may be used to construct the recombinant vector DNAs capable of autonomous replication in cyanobacterium Synechocystis sp. 6803. cells. PMID- 3025719 TI - [Localized mutagenesis of the tetracycline gene in the plasmid pBR322 induced by sodium bisulfite in vitro]. AB - The technique of localized in vitro mutagenesis in the cohesive ends of plasmid pBR322 DNA has been elaborated (separately for BamHI and HindIII sites). Plasmid DNA digested by restriction endonucleases has been treated with sodium bisulphite deaminating cytosine to form uracil in single stranded DNA (cohesive ends of the plasmid). The mutagenized plasmid DNA, free of mutagen, has been treated with bacteriophage T4 ligase. E. coli C600 cells were subsequently transformed by the ligated DNA preparation. The clones having tetracycline gene mutagenized represented 4.0-11.1% and 1.2-3.1% among HindIII and BamHI mutants, respectively, selected as TcR----TcS transformants. Selection of mutagenized DNA by the second endonuclease restriction has increased the mutant yields up to 55.6-78.0% and 10.0-75.4%, respectively. The yield of TcS mutations in the control DNA treated at all stages of experiment, except for mutagen treatment, has reached 0.06% and 0.2%, respectively. PMID- 3025720 TI - [The use of transposons for the mutagenesis of R. phaseoli and R. japonicum]. AB - Plasmid pSUp2011 has been used to transfer transposon Tn5 into the cells of R. japonicum 110 and R. phaseoli 693. Transposition of Tn5 into the chromosomes of R. japonicum and R. phaseoli has been demonstrated,resulting in isolation of auxotrophic and symbiotic mutants of both strains. The frequencies of selected auxotrophic mutations have reached 4% in R. japonicum 110 and 0.6% in R. phaseoli 693. Streptomycin resistance gene locating on Tn5 has been found to be phenotypically expressed in R. japonicum 110 and R. phaseoli 693 cells. PMID- 3025721 TI - [Characteristics of the genome of poliovirus type 3 isolated from patients with poliomyelitis in Moldavia]. AB - Twelve poliovirus isolates of serotype 3 from patients with paralytic poliomyelitis have been analyzed by oligonucleotide mapping of the viral genomes. All the studied strains were isolated from patients in different regions of the Moldavian SSR in 1982. The maps of all isolates are similar but they do not practically possess any large oligonucleotides characteristic of the vaccine strain of type 3 poliovirus. It is concluded that a wild neurovirulent strain of type 3 poliovirus, that circulated in 1982 in the Moldavian SSR was the cause of paralytic poliomyelitis cases. All the studied isolates are suggested to have been derived relatively recently from the common ancestor. PMID- 3025722 TI - Mutagenic DNA repair in Escherichia coli. XIII. Proofreading exonuclease of DNA polymerase III holoenzyme is not operational during UV mutagenesis. AB - We have introduced a mutD5 mutation (which results in defective 3'-5'-exonuclease activity of the epsilon proofreading subunit of DNA polymerase III holoenzyme) into excision-defective Escherichia coli strains with varying SOS responses to UV light. MutD5 increased the spontaneous mutation frequency in all strains tested, including recA430, umuC122::Tn5, and umuC36 derivatives. It had no effect on UV mutability or immutability in any strain or on misincorporation revealed by delayed photoreversal in UV-irradiated umuC36, umuC122::Tn5, or recA430 bacteria. It is concluded that the epsilon proofreading subunit of DNA polymerase III holoenzyme is excluded, inhibited, or inoperative during misincorporation and mutagenesis after UV. PMID- 3025723 TI - Biological and biochemical consequences of the human ERCC-1 repair gene after transfection into a repair-deficient CHO cell line. AB - The consequences of the presence of the human gene ERCC1 in repair-deficient 43 3B cells were examined. The gene restores the sensitivity of this mutant not only to UV but also to 4NQO, N-Ac-AAF and alkylating agents to the normal level. Also, the frequency of mutation induction by UV at the Na+/K+-ATPase locus returns to the level of CHO wild-type cells. Additionally, the rate of cyclobutane pyrimidine dimer removal approaches that in wild-type CHO cells. The results obtained indicate that the human gene ERCC-1 restores the impaired functions in 43-3B, and that the gene is probably functionally homologous to the defective one in the 43-3B cell line. Some evidence was found for a difference between the human gene product and its rodent counterpart, as the restoration of normal sensitivity to 4NQO, ENU and N-Ac-AAF was complete whereas it was not for UV. PMID- 3025724 TI - A cell cycle-associated pathway for repair of DNA-protein crosslinks in mammalian cells. AB - Bulky adducts to DNA including DNA-protein crosslinks formed with trans platinum(II)diammine-dichloride are repaired largely by the nucleotide excision pathway in mammalian cells. The discovery in this laboratory that cells deficient in nucleotide excision repair, i.e., SV40-virus transformed SV-XP20S cells, can efficiently repair DNA-protein crosslinks implicates a second pathway. In this report, details concerning this pathway are presented. DNA-protein crosslinks induced with 20 microM trans-platinum were assayed by the membrane alkaline elution procedure of Kohn. DNA replication was measured by CsCl gradient separation of newly synthesized DNA that had incorporated 5-bromodeoxyuridine. The following results indicate that this new repair pathway is associated with cell cycling: Whereas rapidly proliferating human cells deficient in excision repair (SV40 transformed XP20S, group A) are proficient in repair of DNA-protein crosslinks, the more slowly growing untransformed parent line is deficient but can complete repair after prolonged periods of 4-6 days, the approximate doubling time of the cell population. Either "used" culture medium or cycloheximide (1 microgram/ml) inhibits cell proliferation, protein synthesis, DNA replication and crosslink repair. In the presence of increasing concentrations of cycloheximide (0.01-5 micrograms/ml) the percent of DNA replication decreases and is essentially equivalent to the percent of crosslink repair. The following results indicate that this new repair pathway, though associated with cell cycling, is independent of DNA replication per se. The rates of DNA-protein crosslink repair and DNA replication are essentially the same in mouse L1210 cells rapidly proliferating in 20% serum supplement; however, to slower proliferation rates in 1% serum rate of crosslink repair is slower but differs from that of DNA replication. In the presence of aphidicolin (10 micrograms/ml) cells can repair DNA-protein crosslinks in virtually the complete absence of DNA replication, though the rate is slower in both nucleotide excision-proficient and -deficient cells. Thus, DNA replication is not essential for repair of DNA-protein crosslinks. Comparison of the kinetics of replication and DNA-protein crosslink repair of pulse-labeled indicates that, in the absence of metabolic inhibitors, repair of the crosslinks is independent of replication per se and, therefore, DNA recombination events are not involved in this repair process. We conclude, therefore, that the new repair pathway is not coupled with DNA replication but is with cell cycling. PMID- 3025725 TI - Human parvovirus infection in pregnancy and hydrops fetalis. AB - Human parvovirus is the causative agent of erythema infectiosum, a mild epidemic illness. In a recent outbreak in northeast Scotland, six women had serologic evidence of having contracted human parvovirus infection during pregnancy. Two of the women had midtrimester abortions, and both abortuses were grossly hydropic with anemia. They had similar microscopical histopathological features--a pronounced leukoerythroblastic reaction, hepatitis, excessive iron pigment in the liver, and eosinophilic changes in the hematopoietic cell nuclei. Dot hybridization with radiolabeled human parvovirus DNA probes revealed viral DNA in several tissues from both fetuses, indicating that they had been infected by the virus in utero. The remaining four women had uncomplicated pregnancies and delivered apparently healthy babies, none of whom had human parvovirus-specific IgM antibody at delivery. We conclude that this common virus may pose a serious risk to the fetus after maternal infection. PMID- 3025726 TI - Corticotropin-releasing hormone in the hypercortisolism of depression and Cushing's disease. PMID- 3025727 TI - Low risk of herpes simplex virus infections in neonates exposed to the virus at the time of vaginal delivery to mothers with recurrent genital herpes simplex virus infections. AB - We studied the risk of herpes simplex virus (HSV) infections in neonates exposed to HSV at the time of vaginal delivery to mothers with a history of recurrent genital HSV infections. None of 34 infants exposed to HSV type 2 acquired an HSV infection. On the basis of this sample, the 95 percent confidence limit for the theoretical maximum infection rate is 8 percent. Cord blood or blood obtained during the first two weeks of life was available from 33 of the 34 exposed, uninfected neonates. All 33 of the samples possessed demonstrable neutralizing antibody to HSV type 2, and 79 percent had titers above 1:20. These results were compared with those in a previously studied group of neonates with HSV infections; the latter infants were significantly less likely at the onset of symptoms to have demonstrable neutralizing antibody to HSV type 2 (P = 0.000148) or to have titers above 1:20 (P less than 0.00001). We conclude that given the low attack rate, empirical antiviral therapy is not warranted in all infants of mothers with recurrent genital HSV infection who are exposed to the virus in the birth canal. Our findings suggest that the presence and titer of neutralizing antibody to HSV contribute to the low attack rate. PMID- 3025728 TI - Properties of receptors mediating opioid effects: discrimination of receptor types. AB - Quantitative characterization of opioid receptor types is now feasible by several methods which examine different aspects of ligand interaction with receptor and the induction of agonist effect. Application of these approaches to opioid receptor classification, using receptor-type selective agonists and antagonists, should lead to the development of profiles of the properties of each type of receptor which will be useful in defining the receptors mediating opioid action in other systems. PMID- 3025729 TI - Opiate receptor subtypes and brain function. PMID- 3025730 TI - Kappa isoreceptors: neuroendocrine and neurochemical evidence. AB - It has become clear, in the area of opiate research, that there are multiple opioid receptors and endogenous ligands. The functional roles of these opioid systems are being established as these concepts evolve and as our analytical capabilities become more sophisticated and thorough. When complete dose-response studies are performed and a number of antagonists are examined, a clearer definition of isoreceptor populations may also occur. Such appears to be the case with kappa agonists, which may in fact comprise two groups of compounds (table 1). This separation into two groups has also been observed in the dog cerebral artery (table 1) (Altura et al. 1984). Taken together, these data are sufficient to propose the testable hypothesis that there are kappa isoreceptor populations in the brain that probably subserve a vast array of discrete and different regulatory functions. PMID- 3025731 TI - Multiple receptor types in opioid discrimination. PMID- 3025732 TI - Neurobiological substrates of drug self-administration. PMID- 3025733 TI - Neurochemical substrates for opiate reinforcement. AB - The studies reported herein summarize our work to date aimed at determining the neurochemical substrates for the reinforcing properties of opiates. Rats were trained to self-administer heroin intravenously in daily 3-hour sessions, and pharmacological blockade and neurotoxin-induced lesions were used to define the neurochemical substrates for this reinforcing action. Low-dose DA receptor blockade failed to alter heroin self-administration but significantly increased cocaine self-administration, presumably reflecting a decrease in the reinforcing effectiveness of cocaine. Destruction of presynaptic DA terminals within the N.Acc. produced extinction of cocaine, but not heroin, self-administration. Opiate receptor blockade with systemic naloxone increased heroin, but not cocaine, self-administration. Methylnaloxonium injections into the N.Acc. were effective in increasing heroin self-administration at doses one-eighth those observed for intracerebroventricular injections. Reinforcement has been explored using a place-preference procedure and a self-administration drug-substitution paradigm. Mu/delta agonists such as B-END readily produce a naloxone-reversible place preference. Fentanyl derivatives also produce place preference and substitute for heroin during self-administration. The kappa agonist U50-488 produces place aversion, not place preference, and does not readily substitute for heroin. Altogether, these results suggest that mu/delta receptor subtypes in the region of the N.Acc. may be an important neurochemical substrate for opiate reinforcement. PMID- 3025734 TI - Opiate receptor subtypes associated with the brain mechanisms of feeding and reward. PMID- 3025735 TI - In vivo interactions among opiate receptor agonists and antagonists. PMID- 3025736 TI - Selective radioligands for characterization and neuroanatomical distribution studies of brain opioid receptors. PMID- 3025737 TI - Opiate-inhibited adenylate cyclase in mammalian brain membranes. PMID- 3025738 TI - Modulation of opioid peptide metabolism by seizures: differentiation of opioid subclasses. AB - Until now, we have measured dynorphin-ir and enkephalin-ir at only a few time points after a single seizure or after multiple seizures in most of the models we have employed. Except for the genetically seizure-prone gerbil, our data consistently show a transient and robust decrease in dynorphin-ir and a sustained increase in enkephalin-ir in the hippocampal formation subsequent to kainic acid , ECS-, or amygdaloid-kindled convulsive seizures. At this point, kainic acid appears to have the most dramatic effects on hippocampal enkephalin and dynorphin levels, causing an initial decrease followed by a rebound increase beyond control levels, which, for met5-enkephalin, is maintained for at least 2 weeks. Recurrent seizures leading to neurotoxic effects on CA3 pyramidal cells, which are not present after ECS or kindling, may underlie the sustained alteration in enkephalin metabolism after kainic acid. Further investigation into the time course of seizure-induced enkephalin and dynorphin metabolic changes using RIA, ICC, and measurements of opioid mRNA levels may reveal a common pattern of depletion due to immediate release, rebound synthesis according to the severity of demand, and stabilization at a new equilibrium over several days or even weeks in each seizure model. Our preliminary time points suggest striking differences in the rate of metabolism of hippocampal dynorphin and enkephalin in response to seizures. We would like to find out if other perturbations of the hippocampus, primarily the elimination of the influence of its known neurochemical afferents by lesion (as performed on the septohippocampal system described above) or pharmacological blockade, can alter the metabolism of hippocampal opioid peptides and influence subsequent seizure transmission. Distinguishing the physiological conditions that induce metabolic changes in discrete opioid peptidergic pathways may help us to understand how endogenous opioids are involved in the regulation of neuronal excitability in specific brain regions, as well as to understand more about the differential regulation of opioid peptide metabolism in different brain pathways. PMID- 3025739 TI - A superfamily of potentially oncogenic hormone receptors. PMID- 3025741 TI - High frequency of unequal recombination in pseudoautosomal region shown by proviral insertion in transgenic mouse. AB - The mammalian X and Y chromosomes, in contrast to the autosomes, pair during male meiosis only near the telomeres. Alleles localized in this region can undergo reciprocal exchange during meiosis. Because such sequences do not show strict sex linked inheritance, they have been termed pseudoautosomal. In man, several DNA sequences have been described which show pseudoautosomal transmission and which are localized in the pairing region at the ends of the short arms of both the X and Y chromosomes (refs 6-9, and D. Page, unpublished results). We now show that the transgenic mouse strain, Mov-15, contains a single Moloney murine leukaemia virus (M-MuLV) genome in its germline, and genetic evidence indicates that the provirus is integrated into the pseudoautosomal region of the sex chromosome. Proviral copies are lost or gained in 7% of male meioses in this strain, and mouse sequences flanking the provirus are tandemly repeated and highly variable. We conclude that unequal recombination events occur with high frequency in the pairing region, possibly because of the presence of repeated sequences. PMID- 3025740 TI - Predominant use of a V alpha gene segment in mouse T-cell receptors for cytochrome c. AB - The T-cell receptor is a cell surface heterodimer consisting of an alpha and a beta chain that binds foreign antigen in the context of a cell surface molecule encoded by the major histocompatibility complex (MHC), thus restricting the T cell response to the surface of antigen presenting cells. The variable (V) domain of the receptor binds antigen and MHC molecules and is composed of distinct regions encoded by separate gene elements--variable (V alpha and V beta), diversity (D beta) and joining (J alpha and J beta)--rearranged and joined during T-cell differentiation to generate contiguous V alpha and V beta genes. T-helper cells, which facilitate T and B cell responses, bind antigen in the context of a class II MHC molecule. The helper T-cell response to cytochrome c in mice is a well-defined model for studying the T-cell response to restricted antigen and MHC determinants. Only mice expressing certain class II molecules can respond to this antigen (Ek alpha Ek beta, Ek alpha Eb beta, Ev alpha Ev beta and Ek alpha Es beta). Most T cells appear to recognize the C-terminal peptide of cytochrome c (residues 81-104 in pigeon cytochrome c). We have raised helper T cells to pigeon cytochrome c or its C-terminal peptide analogues in four different MHC congenic strains of mice encoding each of the four responding class II molecules. We have isolated and sequenced seven V alpha genes and six V beta genes and analysed seven additional helper T cells by Northern blot to compare the structure of the V alpha and V beta gene segments with their antigen and MHC specificities. We have added five examples taken from the literature. These data show that a single V alpha gene segment is responsible for a large part of the response of mice to cytochrome c but there is no simple correlation of MHC restriction with gene segment use. PMID- 3025743 TI - Molecular cloning and polymorphism of the human immune deficiency virus type 2. AB - We recently reported the isolation of a novel retrovirus, the human immune deficiency virus type 2 (HIV-2, previously named LAV-2), from patients with acquired immune deficiency syndrome (AIDS) originating from West Africa. This virus is related to HIV-1, the causative agent of the AIDS epidemic now spreading in Central and East Africa, as well as the USA and Europe (see ref. 3 for review) both by its morphology and by its tropism and in vitro cytopathic effect on CD4 (T4) positive cell lines and lymphocytes. But preliminary hybridization experiments indicated that there are substantiated differences between the sequences of the two genomes. Furthermore, the proteins of HIV-1 and HIV-2 have different sizes and their serological cross-reactivity is restricted to the major core protein, as the envelope glycoproteins of HIV-2 are not immunoprecipitated by HIV-1-positive sera. We now report the molecular cloning of the complete 9.5 kilobase (kb) genome of HIV-2, the observation of restriction site polymorphism between different isolates, and a preliminary analysis of the relationship of HIV 2 with other human and simian retroviruses. PMID- 3025742 TI - Steroid-free glucocorticoid receptor binds specifically to mouse mammary tumour virus DNA. AB - Steroid hormones are thought to modulate gene expression through their interaction with receptor proteins. The intracellular localization of unoccupied receptor proteins has been a subject of controversy: free glucocorticoid receptor appears to reside in the cytoplasm and moves to the cell nucleus only after binding the steroid. The purified hormone-bound glucocorticoid receptor has been shown to bind selectively to hormone regulatory elements (HRE) in the vicinity of hormonally-inducible promoters and, in particular, in the long terminal repeat (LTR) region of mouse mammary tumour virus (MMTV). We have tackled the question of whether the hormone itself is required for the interaction of the receptor protein with the HRE. Using monoclonal antibodies to the receptor we find that upon heat-activation the steroid-free glucocorticoid receptor present in rat liver cytosol binds specifically in vitro to the HRE of MMTV. No qualitative differences in the DNaseI-footprints were detected when hormone-free receptor was compared to the hormone-receptor complex or even receptor complexed with the hormone antagonist RU486. We conclude that the steroid ligand is not an absolute requirement for generating the conformation of the glucocorticoid receptor that allows its interaction with the HRE in vitro. An alternative function of the hormone in vivo could be to modulate nuclear partitioning of the receptor. PMID- 3025744 TI - New approaches to vitamin D. PMID- 3025745 TI - Mitochondrial DNA and human evolution. AB - Mitochondrial DNAs from 147 people, drawn from five geographic populations have been analysed by restriction mapping. All these mitochondrial DNAs stem from one woman who is postulated to have lived about 200,000 years ago, probably in Africa. All the populations examined except the African population have multiple origins, implying that each area was colonised repeatedly. PMID- 3025746 TI - Multiple classes of glutamate receptor on depolarizing bipolar cells in retina. AB - Multiple subtypes of excitatory amino acid receptor have been found on individual dissociated neurones. These findings were obtained from cells without intact synaptic connections, so the functional roles for such receptor subtypes are unknown. We have recorded intracellular responses from depolarizing bipolar cells (DBC) that receive direct synaptic input from two distinct populations of neurones: rods and cones. We report here that 2-amino-4-phosphonobutyrate (APB), a glutamate analogue, reveals two subtypes of glutamate receptors on DBCs. APB acts on the same receptor that mediates synaptic transmission from rods but has no action on the second subtype of glutamate receptor. These results show that the rod and cone inputs to DBCs are mediated by pharmacologically distinct receptors and that subtypes of glutamate receptor existing on single neurones can subserve separate, functionally defined synaptic inputs. PMID- 3025747 TI - A new mechanism for induced vitamin D deficiency in calcium deprivation. AB - Synthesis of vitamin D in the skin in response to ultraviolet light is the main determinant of vitamin D status in man and it is therefore surprising that rickets and osteomalacia, clinical signs of vitamin D deficiency, remain common in tropical and subtropical countries. Skin pigmentation can reduce vitamin D formation but this is a negligible limitation in people exposed to abundant ultraviolet light. Earlier studies in animals and man suggested that another environmental factor, the low calcium/high cereal diet typical of susceptible populations, might affect the efficiency of vitamin D utilization. We show here in rats that the rate of inactivation of vitamin D in the liver is increased by calcium deprivation. The effect is mediated by 1,25-dihydroxyvitamin D, produced in response to secondary hyperparathyroidism, which promotes hepatic conversion of vitamin D to polar inactivation products that are excreted in bile. This finding has widespread implications both for understanding the pathogenesis of endemic rickets and in that it provides a unifying mechanism for the development of vitamin D deficiency in many clinical disorders. PMID- 3025748 TI - Herpes simplex virus type-1 can reactivate transcription of latent human immunodeficiency virus. AB - The human immunodeficiency virus, HIV (formerly T-cell lymphotropic virus type III, HTLV-III or lymphadenopathy-associated virus, LAV) is the primary cause of AIDS (acquired immune deficiency syndrome). Patients with AIDS have profound immunosuppression as a result of almost complete absence of the OKT4+ cell population and are predisposed to a number of opportunistic infections as well as to certain malignant diseases such as Kaposi's sarcoma and B-cell tumours. The majority of the opportunistic infections observed in AIDS patients are from the herpesvirus group and these are frequently the cause of death in AIDS patients. We have therefore investigated the effect of herpes virus infection on the expression of HIV and we provide evidence that herpes simplex virus type I (HSV I) can reactivate transcription of latent HIV. In OKT4+ human T-cells HIV replicates to high virus titres, resulting in high level expression of viral RNA. This high level of expression has been attributed to virus-associated trans acting factors that increase gene expression, directed by the HIV long terminal repeats (LTR), post-transcriptionally. In our studies we have tested whether transcription directed by the LTR of HIV is stimulated by HSV-I. PMID- 3025750 TI - Oestradiol induction of a glucocorticoid-responsive gene by a chimaeric receptor. AB - Steroid hormone receptors are a class of cell-specific trans-acting transcription regulatory factors whose activity is controlled by specific binding of the hormone. The hormone-receptor complex appears to associate with promoter/enhancer elements of specific target genes, resulting in activation of transcription (see refs 1 and 2 for reviews). Sequence comparison between the oestrogen, glucocorticoid and progesterone receptors (refs 7, 8 and unpublished results) and site-directed mutation analysis, has identified in each at least two functional domains important for steroid receptor function. Region E (Fig. 1a), is the hormone-binding domain; region C is a 66-amino-acid region (Figs 1a,b) that is more highly conserved than the hormone-binding domain and has the potential to form at least two zinc-stabilized 'DNA-binding fingers' analogous to those proposed for the Xenopus transcription factor TFIIIA. We and others have suggested that this region may be the receptor's DNA-binding domain. We show here that point mutations replacing two cysteines by two histidines in the first potential DNA-binding finger of the human oestrogen receptor (hER) prevent it from activating gene transcription. We further show that a chimaeric receptor formed by replacing this 66-amino-acid region of the hER with that of the human glucocorticoid receptor (hGR) activates expression of a glucocorticoid-inducible gene, but not of an oestrogen-inducible gene, in the presence of oestradiol. Thus, region C determines the receptor's specificity for target genes. PMID- 3025749 TI - Bovine papillomavirus E2 trans-activating gene product binds to specific sites in papillomavirus DNA. AB - Enhancers are cis-acting elements that activate transcription in higher eukaryotes independently of their position or orientation relative to the promoter that they activate. The mechanisms by which enhancers activate transcription are poorly understood, in part because, with the exception of the glucocorticoid receptor, the proteins that directly interact with enhancers have not been purified, nor have the genes encoding them been cloned. The upstream regulatory region (URR) that immediately precedes the early genes of the bovine papillomavirus type 1 genome (BPV) has enhancer activity when it is activated by a trans-acting gene product of the BPV E2 open reading frame (ORF) (Fig. 1). It is not known whether this enhancement represents a direct or indirect effect of E2 on the URR. We have used an E2 peptide expressed in bacteria and a DNA-protein complex immunoprecipitation assay to study E2-mediated enhancement of transcription by the URR. We show here that this peptide directly binds to four specific sites in the BPV URR, and to one site in the human papillomavirus (H)PV16 URR. All the binding sites contain a related sequence of nucleotides; a 23 base pair (bp) fragment containing this sequence can specifically prevent binding of the E2 protein to the BPV URR. The BPV E2-URR enhancer interaction may therefore represent a useful model system for studying the mechanism of transcriptional enhancement, as both an effector protein and its target enhancer can be purified and genetically manipulated. PMID- 3025751 TI - [The surgeon and mass screening for breast cancer]. PMID- 3025753 TI - Active oxygen in methylguanidine synthesis. AB - Methylguanidine (MG), a toxin reported in uremia, is thought to be a product of creatinine oxidation. This study is designed to demonstrate the role of active oxygen in the oxidation of creatinine under conditions compatible with those found in uremia. MG synthesis is moderately stimulated by the superoxide radical derived from 3 mM hypoxanthine and 0.015 units/ml xanthine oxidase and inhibited by the addition of superoxide dismutase. This is increased markedly by the addition of 0.05% hydrogen peroxide and augmented to about 56,000 times the control rate in the presence of hydroxyl radicals derived from the reaction of 10 mM FeSO4 and 0.05% hydrogen peroxide. In addition, MG synthesis is inhibited by the addition of sorbitol, lactulose or ethanol, the scavengers of hydroxyl radicals. These results indicate that creatinine can be oxidized to MG by various species of active oxygen and that one of the mechanisms of MG synthesis is such oxidation. MG, therefore, may be a useful indicator of peroxidation in vivo. PMID- 3025752 TI - Histopathological subclassification of small cell carcinoma of the lung. Evaluation of its prognostic significance. PMID- 3025754 TI - Pharmacokinetics and pharmacodynamics of captopril in patients undergoing continuous ambulatory peritoneal dialysis. AB - Pharmacokinetics and pharmacodynamics of captopril were studied in 5 continuous ambulatory peritoneal dialysis (CAPD) patients (including 2 hypertensive patients) after single oral administration of 50 mg captopril. The pharmacokinetic parameters for plasma free unchanged captopril were time to maximal concentration 1.1 +/- 0.3 h, maximal plasma concentration 387 +/- 75 ng X ml-1, elimination half-life 1.0 +/- 0.3 h, and the area under the concentration time curve 711 +/- 144 ng X h X ml-1. For plasma total captopril (the sum of free unchanged captopril and its disulfide compounds) the values were time to maximal concentration 3.5 +/- 0.6 h and maximal plasma concentration 2,777 +/- 429 ng X ml-1. Captopril was detected in the dialysis fluid in all CAPD patients. Blood pressures in the 2 hypertensive CAPD patients were lower at 24 h after than before captopril administration. These results suggest that captopril may be eliminated by CAPD. In addition, there is a possibility that the antihypertensive effects of captopril may be prolonged in hypertensive CAPD patients. PMID- 3025755 TI - In vitro inhibition of Na-K-ATPase by trace metals: relation to renal and cardiovascular damage. AB - The potential role of trace metals in the pathogenesis of various disease states, especially of renal and cardiovascular disease, has been recognized increasingly. Moreover, altered membrane transport was recently incriminated to play a role in renal tubular syndromes, such as the Fanconi syndrome, as well as in the pathogenesis of volume dependent hypertension. We therefore investigated the possible in vitro effects of various trace metals on Na-K-ATPase, the biochemical correlate of active cellular transmembrane sodium and sodium-dependent transport. To more closely mimic the in vivo situation, we deliberately chose as enzyme source renal tissue homogenate, which may contain protective agents. Under these experimental conditions, the metals studied inhibited the enzyme quantitatively in the following order: Hg greater than Pb greater than Cd greater than Ur greater than Cu greater than Zn greater than Mn greater than Ba greater than Ni greater than Sr. Enzyme kinetic studies showed that Hg, Pb, and Cd competitively, and Cu noncompetitively, inhibited the enzyme. In general, Mg-ATPase was significantly less sensitive to the trace metals. Accumulation of these metals may present serious health hazards by producing a general defect in cell membrane transport. From the other metals studied, i.e., Mn, Ba, Ni and Sr, some may have toxic effects via other mechanisms, whereas some may exert a protective role against toxicity of other agents including metal ions. PMID- 3025756 TI - [Radioisotopic quantification of kidney function using Tc-99m-DMSA. Comparison with creatinine clearance in children with a single kidney]. AB - To assess the accuracy of renal function quantification with Tc 99m-DMSA in children, we compared DMSA renal uptake and creatinine clearance in 16 cases of children with single kidney. The age of the patients ranged from two month to fourteen years. DMSA renal uptake was measured 7 hours after injection and was normalized in percent of the injected activity. A significant correlation was found between creatinine clearance and DMSA uptake (Pearson's r = 0.866, p less than 0.01). Normal creatinine clearance in children (80 to 120 ml/min-1 X 1.73 m 2) allowed determination of normal renal uptake (36 to 60%). This study indicates that in cases of asymmetrical renal impairment renal uptake reflects split renal creatinine clearance. Since the former is much easier to measure, DMSA should play an important role in the evaluation of differential renal function. PMID- 3025757 TI - Alpha-2-adrenergic control of prolactin release. AB - The role of the alpha 2-adrenergic system in regulating prolactin release has been studied in vivo in male rats. Yohimbine administration alone, at doses ranging from 0.2 to 5.0 mg/kg, resulted in a dose-related elevation of plasma prolactin levels from a basal level of 8 +/- 2 to 65 +/- 6 ng/ml at the highest dose. In the same experiment clonidine, 0.2 mg/kg, suppressed basal prolactin levels to 4 +/- 1 ng/ml and returned prolactin levels of all animals receiving 0.2-5.0 mg/kg yohimbine to basal levels. Rats were treated with increasing doses of clonidine (0.05-1.0 mg/kg) in the presence or absence of a constant dose (1.0 mg/kg) of yohimbine. Clonidine alone at doses of 0.05 and 0.2 mg/kg again significantly suppressed prolactin levels, while a dose of 1.0 mg/kg did not (failure of high dose clonidine to suppress prolactin levels suggests an additional effect of clonidine on prolactin secretion unrelated to alpha 2 adrenergic agonist action). All three doses of clonidine completely reversed yohimbine-induced prolactin release. Basal prolactin levels were also significantly reduced by the selective alpha 2-adrenergic agonist UK-14,304 at a dose of 0.2 mg/kg. Yohimbine-induced prolactin release was reversed by UK-14,304 at doses of 0.2 and 1.0 mg/kg, but not at the lowest dose studied, 0.05 mg/kg. The lower potency of UK-14,304 than clonidine in this assay is consistent with the lower potency of UK-14,304 as an alpha 2-adrenergic-agonist antihypertensive agent. Several alpha 2-antagonists in addition to yohimbine were studied.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3025758 TI - Effect of catechol estrogens on the preovulatory content of luteinizing hormone releasing hormone in the median eminence of the rat. AB - Cycling rats injected with 2-hydroxyestrone or 2-hydroxyestradiol at 09.00 or 10.00 h in the morning of proestrus do not express the normal preovulatory LH surge in the afternoon of the day. The LHRH content of the median eminence in control animals decreases sharply in the afternoon from elevated noon and morning levels. The catechol estrogen-treated rats fail to show the decrease. Thus the catechol estrogens block the LH surge at its usual time by influencing the changes in the concentration of LHRH in the median eminence on proestrus. Since the catechol estrogens have short biological half-lives, their effect on the LHRH content in the afternoon must originate in the morning at the time of the endogenous estradiol (E2) peak. These results have implications in the physiological processes responsible for the positive feedback of estradiol on the preovulatory LH surge in the rat. PMID- 3025759 TI - Reversal of the delta-9-tetrahydrocannabinol inhibitory effect on prolactin secretion by rostral deafferentation of the medial basal hypothalamus. AB - The effect of rostral deafferentation of the medial basal hypothalamus (MBH) on delta-9-tetrahydrocannabinol (THC)-induced changes in serum prolactin (PRL) concentrations was investigated in female rats having retrochiasmatic frontal cuts that transected the rostral hypothalamus. Cuts dorsal to the hypothalamus were produced in the same plane in other animals in order to control for possible effects of the surgical procedure or dorsal brain damage. All animals were ovariectomized 28-35 days after stereotaxic surgery to obviate potential confounding effects of differences in ovarian function between groups. Unlesioned rats were ovariectomized to provide a positive control group for THC inhibitory activity. At least 4 weeks after ovariectomy, animals were treated intravenously with THC (0.5 or 1.0 mg/kg body weight) or vehicle at the midpoint of a 110-min experimental period during which blood samples were obtained at 10-min intervals via indwelling atrial cannulae. Serum PRL concentrations were determined by radioimmunoassay and cut locations were confirmed histologically. When administered to ovariectomized animals without brain lesions, THC suppressed serum PRL concentrations from the average treatment level within 30 min (p less than 0.05), and PRL levels remained suppressed for the remainder of the posttreatment sampling period. Treatment with the vehicle alone was without effect. Animals with retrochiasmatic plane cuts that did not transect the rostral hypothalamus similarly displayed PRL suppression in response to THC administration (p less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3025760 TI - Control of ACTH secretion by the central nucleus of the amygdala: implication of the serotoninergic system and its relevance to the glucocorticoid delayed negative feedback mechanism. AB - The possible implication of the amygdaloid central nucleus (ACE) of the rat in the control of ACTH secretion in response to immobilization stress was assessed. The ACTH secretion, in response to stress and/or bilateral lesions of the ACE, was correlated with the serotoninergic activity in specific hypothalamic and amygdaloid nuclei. Bilateral lesions of the ACE produced a striking decrease of plasma ACTH levels in response to stress. However, basal plasma ACTH levels measured between 7 and 11 a.m. were identical in both control and lesioned groups. Stress, applied to intact animals, did not modify the serotoninergic activity in any of the following areas: hypothalamic paraventricular (PVH), ventromedial (VMH) and dorsomedial (DMH) nuclei; the anterior hypothalamic area (AHA); the lateral part of the basal amygdaloid nucleus (ABL), the amygdaloid medial (AME) and cortical (ACO) nuclei. However, lesion of the ACE increased the serotoninergic activity in all these structures except for the VMH. Immobilization stress applied to lesioned animals decreased the serotoninergic activity to control levels in the PVH, AHA and DMH and decreased the activity to below control levels in the VMH. The serotoninergic activity remained at an increased level in the glucocorticoid receptor-rich areas of the amygdala, namely the AME, ACO and ABL nuclei. These results provide evidence for a stimulatory role of the central nucleus of the amygdala in the control of ACTH secretion. Moreover, they substantiate an implication of the amygdaloid complex in the control of the delayed negative feedback of glucocorticoids on ACTH secretion via interaction with the serotoninergic system. PMID- 3025761 TI - Activation of the CNS noradrenergic system may inhibit as well as facilitate pituitary luteinizing hormone release. AB - Earlier work established that neural secretion of luteinizing hormone-releasing hormone (LH-RH) and the resultant release of pituitary gonadotropins could be facilitated by activating alpha-receptors of a central noradrenergic (NA) system. The present study emphasizes that central NA mechanisms may also inhibit LH release largely through activation of beta-adrenergic receptors. PMID- 3025762 TI - Acute infusion of lithium chloride raises blood pressure in the conscious rat. AB - The effects of acute infusion of lithium chloride (LiCl) were studied on mean arterial pressure (MAP) and magnocellular activity as shown by the concentrations of vasopressin-associated neurophysin ([VP-RNP]) and concentrations of oxytocin associated neurophysin ([OT-RNP]) in plasma in conscious Long-Evans rats. Chronically-cannulated rats were infused intravenously at 10 microliter/100 g body wt/min with 13% LiCl for 20 min (total dose = 6.16 mequiv./kg body wt) or 0.65% LiCl for 60 min (total dose = 0.92 mequiv./kg body wt). Effects of 13% LiCl on mean arterial pressure were also examined in vasopressin-deficient homozygous Brattleboro rats. For Long-Evans rats, infusion of 13% LiCl produced rapid and significant (P less than 0.001) increases in mean arterial pressure, the concentration of lithium in plasma ([Li+]), plasma osmolality (Posmol), [VP-RNP], [OT-RNP] and significant decreases in heart rate and sodium concentration in plasma ([Na+]). For similar changes in plasma osmolality, lithium had a greater effect than sodium on mean arterial pressure, [VP-RNP], [OT-RNP]. For the 20 min infusion of 13% LiCl, there was a significant relationship (P less than 0.033) between delta MAP and log delta[VP-RNP] with a slope of 11.9 mmHg fmol-1 ml-1 (r = 0.5678). Unlike that of Long-Evans rats, infusion of 13% LiCl only did not produce significant changes of mean arterial pressure in Brattleboro rats. For Long-Evans rats, infusion of 0.65% LiCl resulted in more gradual and smaller elevations of blood pressure, [Li+] and smaller decreases in heart rate with no significant changes in plasma osmolality, [Na+], [VP-RNP] and [OT-RNP].(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3025763 TI - Novel GABA analogues as hypotensive agents. AB - The actions of four analogues of gamma-aminobutyric acid (GABA) on blood pressure and heart rate were measured in the cat after intracerebroventricular administration. These compounds were previously found to inhibit binding to GABAA receptors of neuronal membranes from the CNS of the rat. Each of the drugs, together with GABA, produced an average maximum reduction in blood pressure of 27.63% +/- 12.5. However, aminoethanethiosulfonic acid (AETS) was the most potent (ED50 = 2.24 X 10(-10) mol/kg) of the drugs, followed by 5-phenyl-2-pyrrole propionic acid (PPP), urocanic acid (UCA), m-aminobenzoic acid (MABA) and GABA. None of the compounds produced a significant effect on heart rate. The fact that these analogues mimicked the action of GABA on the cardiovascular system of the cat and that they were able to inhibit binding to GABAA receptors, indicates that they may be GABA agonists. PMID- 3025764 TI - Interactions between three pyridazinyl-GABA derivatives and central GABA and glycine receptors in the rat, an in vivo microiontophoretic study. AB - Three pyridazinyl-gamma-aminobutyric acid (GABA) derivatives, SR 95103, SR 42641 and SR 95531, have previously been shown to be specific, competitive and reversible GABAA antagonists. For all three compounds selectivity was claimed mainly on the basis of biochemical results. However the absence of an interaction with the binding site for strychnine does not preclude an interaction with the glycine receptor. Therefore the interaction between these compounds and GABA- and glycine-elicited responses in the rat cuneate nucleus was examined in vivo by microiontophoretic techniques using extracellular recordings. Preliminary experiments in the somesthetic cortex of the rat (18 cells) confirmed that SR 95103 (100 mM; 0-100 nA) blocked GABA-evoked responses. In the cuneate nucleus SR 95103 (100 mM), SR 42641 (5 mM), SR 95531 (5 mM) and bicuculline methochloride (BMC; 5 mM) blocked GABA-elicited responses in a dose-dependent, competitive and reversible fashion for ejection currents up to 100 nA. The compound SR 95103 appeared to be less potent than bicuculline and also antagonized glycine-induced responses. Both SR 42641 and SR 95531 appeared to be equipotent to bicuculline and selective for the GABA receptor. Based on these results, it is postulated that SR 42641 and SR 95531 are potent and reversible GABAA antagonists and could be useful tools to further characterize the GABA receptor. PMID- 3025765 TI - Effects of delta-9-tetrahydrocannabinol, 11-hydroxy-delta-9-tetrahydrocannabinol and cannabidiol on neuromuscular transmission in the frog. AB - Intracellular recording techniques were used on neuromuscular junctions of the sartorius muscle of the frog, in vitro, to define the synaptic pharmacology of delta-9-tetrahydrocannabinol (THC), 11-hydroxy-THC and cannabidiol (CBD). The frequency of miniature endplate potentials was increased by THC, decreased by CBD and was unaffected by 11-hydroxy-THC, whereas the amplitude of the miniature endplate potentials was depressed by all three cannabinoids. In addition, the mean quantum content of the endplate potential (m) was first increased and then decreased by THC and 11-hydroxy-THC, but CBD produced only depression. Changes in m and the frequency of the miniature endplate potential indicated presynaptic sites of drug action and reduction of the amplitude of the miniature endplate potential suggested a postsynaptic site. The findings suggest possible mechanisms of action for the central excitatory and depressant properties of the cannabinoids. PMID- 3025766 TI - Specific binding of a phenyl-pyridazinium derivative endowed with GABAA receptor antagonist activity to rat brain. AB - SR 95531 has been shown to be a potent, selective, reversible and competitive GABAA antagonist. In the present study we report that (3H)SR 95531 binds with high affinity and in a specific and saturable manner to rat brain membranes. Scatchard analysis revealed two binding sites (KD: 6 nM; Bmax: 0.24 pmol/mg protein and KD: 38 nM; Bmax: 0.66 pmol/mg protein). Only GABA ligands were effective displacers of (3H)SR 95531. The respective IC50 values obtained with these compounds suggests that (3H)SR 95531 labels the GABA receptor in its antagonist conformation. PMID- 3025767 TI - The pharmacokinetic profile of lithium in rat and mouse; an important factor in psychopharmacological investigation of the drug. AB - The pharmacokinetic characteristics of lithium and the profile of plasma lithium concentration at steady state in both the mouse and the rat have been determined. The half life of lithium in both rodents was shorter (3.5 h and 6 h) than that found during maintenance therapy in man. Following a loading dose (10 mmol/kg s.c.) and twice daily maintenance injections (3 mmol/kg s.c.) of lithium chloride the plasma concentration remained above the accepted human therapeutic minimum (0.4 mM) for 16 of every 24 h in the mouse and throughout the entire 24 h period in the rat. Maximum concentrations in both species were below the range at which toxic effects might be expected to occur. PMID- 3025768 TI - Effect of enkephalins on the function of calcium-regulating endocrine glands. AB - Experiments on rats have shown that administration of a synthetic analog of leucine enkephalin at a dose of 100 mg/kg did not change the content of the immunoreactive parathyroid hormone in the blood plasma, but eliminated the parathyroidin-stimulating effect on the cAMP concentration in the renal tissue and AP activity in the blood. We conclude that enkephalins do not influence the secretion and metabolism of the parathyroid hormone, but block its peripheral effects and stimulate thyroid C-cell function. PMID- 3025769 TI - Taurine interferes with spiperone binding in the striatum. AB - The effects of taurine and its structural analogues and two new anticonvulsant derivatives, taltrimide and MY-103, on the function of brain dopaminergic systems were studied by assessing their interference with the binding of [3H]spiperone to synaptic membranes isolated from rat striata. Two populations of binding sites were detected. The binding was effectively displaced by (+)butaclamol and dopamine, serotonin being less potent by one order of magnitude. Taurine, taltrimide and MY-103 inhibited spiperone binding moderately. The binding constants of both high- and low-affinity components and the maximal binding capacity of the low-affinity component decreased in the presence of taurine. The results show that taurine and its novel anticonvulsant derivates could modulate the function of striatal dopaminergic systems. PMID- 3025770 TI - Autoradiographic localization in rat brain of kappa opiate binding sites labelled by [3H]bremazocine. AB - [3H]Bremazocine, in the presence of saturating concentrations of mu and delta receptor blocking agents, was used to label putative kappa opiate binding sites in rat brain. The binding of [3H]bremazocine under these conditions was completely displaced with high affinity by U-50488H and dynorphin1-17, and the potency of a series of opiate ligands was consistent with an action at kappa receptors. Therefore, [3H]bremazocine, in the presence of mu and delta blockers, was used to localize U-50488H-displaceable kappa binding sites by autoradiography. A distribution different from that of mu and delta receptors was seen, with levels highest in the claustrum, striatum, medial preoptic area, suprachiasmatic nucleus, medial amygdala and superior layer of the superior colliculus. The results show that the U-50488H-displaceable kappa sites have a distinct distribution which is discussed in terms of the possible functional roles of kappa receptors. PMID- 3025771 TI - Distribution of opioid binding sites in the guinea pig hippocampus as compared to the rat: a quantitative analysis. AB - In vitro autoradiography of cryostat sections revealed major differences between the distribution of opioid binding sites in the hippocampus of the guinea pig and the rat. Only very low binding was found in the pyramidal cell layer, the dentate granular cell layer and the commissural-associational zone of the dentate molecular layer of the guinea pig, whereas these areas were moderately to densely labeled in the rat. In the guinea pig an enrichment of sites was observed in the terminal field of the mossy fiber system in the hilus which was absent in the rat. Binding sites in the guinea pig were found to be mainly of the kappa and mu type. The distribution of [Leu]enkephalin immunoreactivity does not correlate well with the distribution of delta opioid binding sites in the hippocampus. Quantification of opioid binding sites in the hippocampus demonstrates that no one type of site can be assigned to a specific hippocampal subregion nor does the intensity or the pattern of distribution of binding types agree well with the distribution of endogenous opioid peptides. PMID- 3025772 TI - Cytomegalovirus encephalitis associated with episodic neurologic deficits and OKT 8+ pleocytosis. AB - After 3 days of symptoms suggesting a viral illness, a 35-year-old man experienced three episodes of aphasia, right-sided sensory symptoms, and bifrontal headache. Each lasted several hours. CSF examination revealed a moderate lymphocytosis consisting of 80% OKT-8+ cells. Serum anti-cytomegalovirus (anti-CMV) antibody titer was elevated at 1:1,024 and subsequently fell to 1:64. Episodic symptoms recurred 5 months later, at which time the anti-CMV titer peaked at 1:8,192. A trial of inhaled oxygen aborted two episodes after several minutes each. PMID- 3025773 TI - Incidence of antibody to HTLV-I is not increased in Japanese MS patients. AB - To examine the association between MS and anti-human T-cell lymphotropic virus-I (HTLV-I) antibody, we studied serum and CSF antibody to HTLV-I in 27 Japanese MS patients by an indirect immunofluorescence method sensitive and specific enough to detect carriers of HTLV-I. The antibody was detected in 3 of 27 MS patients (11.1%), in 4 of 48 patients (8.3%) with other neurologic diseases, and in 8.3% of 2,500 healthy blood donors. There was no significant difference in the incidence between the three groups. The titer of the antibody was low in CSF when compared with that in serum in all seropositive MS patients. Fluctuations in the CSF antibody titer were not observed in any of 14 MS patients sampled repeatedly. PMID- 3025774 TI - Peripheral neuropathy and light chain myeloma: case report. PMID- 3025775 TI - Mitochondrial encephalomyopathy and partial cytochrome c oxidase deficiency. AB - A 52-year-old man had slowly progressive weakness and wasting of limb-muscles, sensorineural hearing loss, and complex partial seizures. CT showed cerebral atrophy, but he was not demented. Muscle biopsy showed ragged-red fibers and decreased histochemical stain for cytochrome c oxidase. Biochemical studies showed decreased cytochrome c oxidase activity in crude muscle extracts and in isolated mitochondria (44 and 30% of normal), while other mitochondrial enzymes were normal. A comparable decrease of immunologically reactive enzyme protein was shown by immunotitration with antibodies against human heart cytochrome c oxidase. Partial defects of cytochrome c oxidase may cause adult-onset, slowly progressive mitochondrial encephalomyopathies. PMID- 3025776 TI - Benign reversible muscle cytochrome c oxidase deficiency: a second case. AB - A 6-week-old boy had generalized weakness, requiring assisted ventilation, and lactic acidosis. At 6 months, the lactic acidosis resolved, and the patient started to improve; assisted ventilation was discontinued at 15 months. Muscle biopsies at 4 and 11 months showed accumulation of mitochondria, lipid, and glycogen; cytochrome c oxidase (COX) activity was 11% of the lowest control in the first biopsy and 57% in the second. Immunocytochemistry and immunotitration showed presence of immunologically reactive enzyme protein in both biopsies. This case confirms a previous report of benign infantile myopathy due to reversible COX deficiency. The severe fibrosis in the second biopsy may explain the slower rate of clinical recovery in this child. PMID- 3025777 TI - [Diffuse familial polyposis]. PMID- 3025778 TI - [Therapy of chronic constipation. Observations on the use of a food supplement rich in dietary fiber]. PMID- 3025779 TI - [Cryotherapeutic destruction of invasive tracheo-bronchial tumors. Personal case histories]. AB - Data are presented on 15 cases of invasive tracheobronchial tumours subjected to cryotherapy in 1984-85. The technique is indicated in patients who cannot be given surgical or radiation treatment and in cases of asphyxial syndrome requiring faster deobstruction than is obtainable with radiation treatment. PMID- 3025781 TI - [Malignant mixed mullerian tumors. Presentation of 2 cases and review of the literature]. PMID- 3025780 TI - [Functional compromise of the small airways in subjects exposed to SiO2]. AB - Aims of our study were: to evaluate small airway function of subjects with past or present silica dust exposure and normal spirometric values; to investigate whether small airway disease is related to radiographic signs of silicosis, to cumulative dust exposure (ES) and to cigarette smoking. Maximal expiratory flow at 50% (MEF50) and 25% (MEF25) of forced expired vital capacity were measured in 112 subjects, 69 with radiographic signs of silicosis, group I, and the remaining 43 with normal chest X-rays. Even if age and ES were significantly higher in group I, no significant difference in respiratory function tests and in prevalence of small airway disease was found between the two groups. In both groups small airway function was significantly negatively related to smoking habits, while it was independent of the other variables considered. Multiple regression analysis with MEF50 and MEF25 as dependent variables did not show any significant relationship. We conclude that small airway disease due to encroachment of bronchiolar walls by SiO2 deposition is masqued by the damage produced by cigarette smoking, even in the presence of radiographic signs of silicosis. PMID- 3025782 TI - [Dietary fiber in the prevention and therapy of constipation in pregnancy. A multicenter study]. PMID- 3025783 TI - A potent excitatory input to the nucleus locus coeruleus from the ventrolateral medulla. AB - Our recent anatomic experiments reveal major innervation of the nucleus locus coeruleus (LC) from the nucleus paragigantocellularis (PGi), located in the rostral ventrolateral medulla. In the present studies, low intensity, single pulse electrical stimulation of the PGi synaptically activated most LC neurons examined (69%), while a smaller percentage of LC cells (20%) exhibited pure inhibitory responses. Pharmacologic experiments suggest that the excitatory response may be mediated by an amino acid transmitter. PMID- 3025784 TI - An inborn error of cholesterol biosynthesis. PMID- 3025785 TI - Lead-associated protein in kidney and brain. PMID- 3025786 TI - Fifth disease and arthritis: common immune-mediated responses to parvovirus infection. PMID- 3025787 TI - Human parvovirus and fifth disease. PMID- 3025788 TI - Fifth disease: the frequency of joint involvement in adults. PMID- 3025789 TI - Parenchymal calcification in hepatocellular carcinoma. PMID- 3025790 TI - Oral implantology case reports. PMID- 3025791 TI - Flaviviruses in South Africa: diagnostic procedures. AB - Employing rabbit immune serum, 10 flaviviruses known to be present in South Africa could be divided into 5 serological subgroups. The subgroups conform to the general pattern described for the group. Sera from experimentally infected calves and lambs were monospecific in neutralization tests, but cross-reacted in haemagglutination inhibition tests. These results suggest that sheep and cattle sera from the field can best be tested by microneutralization tests. The greater sensitivity of embryonated hen's eggs for some viruses and of one-day-old mice for other viruses necessitates the employment of both systems for the isolation of flaviviruses from field specimens. PMID- 3025792 TI - Flaviviruses in South Africa: pathogenicity for sheep. AB - Sheep are susceptible to at least 5 of the 10 flaviviruses known to be present in South Africa. Sheep, 7-9 months of age, injected with Wesselsbron, West Nile, Banzi, Uganda-S and AR 5189 (an unidentified virus related to Banzi and Uganda S), responded with a moderate febrile reaction, a low grade viraemia of short duration and the production of virus neutralizing antibodies. The most pronounced manifestations of infection were encountered in pregnant ewes. Infection with West Nile, Banzi and AR 5189 resulted in abortion, stillbirth and neonatal death, characterized by congenital abnormalities of the brain. PMID- 3025793 TI - Detection of human papillomavirus DNA in oral mucosal lesions using in situ DNA hybridization applied on paraffin sections. AB - The in situ DNA hybridization technique, carried out under stringent conditions, was used to detect human papillomavirus (HPV) DNA of types 6, 11, and 16 in paraffin sections of 32 surgically treated oral mucosal lesions. Expression of HPV structural proteins was analyzed by means of the immunoperoxidase (IP-PAP) method. A total of 10 (31.3%) of the 32 lesions proved to express HPV antigens, which were found in 4 of 7 squamous cell papillomas, in 2 of 2 classic condylomas, in 2 of 10 papillary hyperplasias, and in 2 of 3 leukoplakia lesions. Two of the squamous cell papillomas contained HPV 6 DNA, and 4 additional lesions were positive for HPV 11 DNA. In one of the condylomas, a double infection by HPV 6 and 11 was found, while the second was positive for HPV 11 alone. Both the HPV antigen-positive papillary hyperplasias contained HPV 6 DNA, as did the HPV antigen-positive leukoplakia lesions. Of the latter, one was infected by HPV 6 and 11. DNA of the "high-risk" HPV 16 was contained in two lesions: one lichen planus lesion and one of the two squamous cell carcinomas. The results confirm the previously reported evidence of HPV involvement in oral mucosal lesions. The implications of these findings are discussed in terms of the well-established premalignant character of oral leukoplakia and oral lichen planus, although far less commonly versus leukoplakia, with special emphasis on the discovery of the "high-risk" HPV 16 in the latter as well as in oral cancer.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3025794 TI - Experimental sialography: the effects of retrograde infusion of radiographic contrast media on salivary gland morphology and function. A review article. AB - An experimental model for sialography, involving retrograde infusion of contrast medium into the rat submandibular gland under continuous pressure monitoring, was developed. Morphologic alterations induced during infusion of water- and lipid soluble radiographic contrast media and the distribution of the media were examined with light and electron microscopy. An uneven distribution of the media throughout the tissue and variations in the degree of the alterations in different parts of the gland were seen at all stages of filling. The changes primarily consisted of dilation of the luminal system, which successively became more pronounced as infusion continued. Infusion of the lipid-based medium consistently resulted in more severe alterations, including changes in the acini. Tracer studies showed that the water-soluble medium leaked out through the intercellular spaces and to the surrounding tissue, whereas the lipid-based medium appeared to be confined to the luminal space. Ductal filling with either medium resulted in a relatively fast recovery; after prolonged infusion, recovery was slower and additional alterations occurred. The acini showed confluence of secretory granules and vacuole formation. After infusion of the lipid-based compound, autophagic vacuoles and granulomatous alterations developed. Prolonged infusion with either medium resulted in atrophy of the parenchyma and connective tissue proliferation in some glands. An inflammatory infiltrate was present in all glands during the recovery period. Functional alterations, primarily consisting of a decrease in flow rate, were noted after prolonged infusion. The increase in intraluminal pressure induced during infusion was most likely the primary cause of the alterations. In addition, the presence of contrast medium contributed to the long-term effects of the procedure. PMID- 3025795 TI - [Effect of indomethacin on beta adrenergic receptor function in bronchial asthma]. PMID- 3025796 TI - [Primary malignant histiocytoma of the liver]. PMID- 3025797 TI - Juvenile nasopharyngeal angiofibroma. AB - JNA is a highly vascular, benign, yet locally invasive tumor that occurs in preadolescent males. Diagnosis is based on history, physical examination, and radiographic findings. CT scanning is invaluable for evaluating tumor extent. Angiography combined with embolization aids surgeons in identifying the main feeding vessels and decreasing intraoperative blood loss. Surgery is the mainstay of therapy with radiation therapy reserved for inoperable masses. For highly aggressive, recurrent angiofibroma, chemotherapy has shown some promise. PMID- 3025798 TI - Nonsquamous tumors of the nose and paranasal sinuses. AB - Nonsquamous tumors of the nose and paranasal sinuses are an uncommon group of neoplasms. Information is only available from case reports and short series. Therefore, conclusions concerning natural history and response to therapy are uncertain for tumors in these sites. The need for complete and timely evaluation of new and changing clinical signs must be emphasized. For each tumor, maximal interaction among the clinician, radiologist, and pathologist is needed to reach the correct diagnosis and plan effective therapy. This interaction is especially important for the deceptively benign-appearing tumors listed in Table 2. These tumors behave biologically in an unpredictable fashion, and long-term follow-up is mandatory. PMID- 3025800 TI - [Effect of microcrystalline cellulose on the degree of experimental hyperlipidemia]. PMID- 3025799 TI - [Role of adrenoreception in achieving protective reaction to plasmin in the bodies of mammals]. PMID- 3025801 TI - Beta-adrenoceptor function in white blood cells from newborn infants: no relation to plasma catecholamine levels. AB - The maturity of beta-adrenoceptors in newborn infants was studied in relation to the catecholamine surge during labor. Umbilical blood was collected at birth from 12 infants delivered vaginally and 13 infants delivered by elective cesarean section. Granulocytes and lymphocytes were isolated. Receptor numbers and binding affinity were determined in the granulocytes by incubation with 125I iodohydroxybenzylpindolol. Receptor responsiveness was tested by assessing isoproterenol-induced cyclic AMP accumulation in lymphocytes. Significantly higher plasma noradrenaline, adrenaline, and dopamine concentrations were found in infants born vaginally (108; 8.9; 0.9 nmol/liter, liter, respectively, median values) as compared with those delivered by cesarean section (11.0; 2.4; 0.2 nmol/liter). No significant differences in beta-adrenoceptor binding sites (receptor number: 39.2 +/- 2.6 versus 44.7 +/- 5.9 fmol/mg protein and binding affinity: 66.6 +/- 7.8 versus 65.0 +/- 6.2 pM) or responsiveness (maximal isoprenaline induced cAMP formation 52.4 +/- 10.3 versus 40.6 +/- 8.9 pmol/10(6) cells) were found between the two groups of infants. Lymphocyte beta-adrenoceptor sensitivity was similar to that found in adults. The beta-adrenoceptors on whole blood cells seem to be mature at birth and have the same responsiveness as in adults. The higher catecholamine surge during vaginal delivery as compared to elective cesarean section does not seem to affect beta-adrenoceptor function. Our results do not support the idea that reduced beta-adrenoceptor function is the cause of the previously observed inappropriately small cardiovascular and metabolic responses to the exceptionally high plasma catecholamine concentrations at birth. PMID- 3025802 TI - Lymphocyte and granulocyte phosphatidylethanolamine N-methyltransferase: properties and activity in cystic fibrosis. AB - Human lymphocyte and granulocyte membranes contain an enzyme, phosphatidylethanolamine N-methyltransferase (PEMT), which catalyzes the transfer of a methyl group from S-adenosylmethionine to the polar head group of phosphatidylethanolamine to form phosphatidylmonomethylethanolamine. This enzyme, in lymphocyte membranes, has Km for S-adenosylmethionine of 7.01 +/- 2.9 (SD) microM, and specific activity 0.57 +/- 0.31 pmol/mg protein/15 min, is inhibited by S-adenosylhomocysteine, displays optimal activity at pH 8.0-9.0, and is stimulated by isoproterenol in dose-dependent, propranolol-inhibitable fashion, to a lesser extent by epinephrine, but not by norepinephrine, prostaglandin E1, concanavalin A, or adenosine 3':5' cyclic monophosphate, with or without phosphodiesterase inhibitors. Granulocyte membrane PEMT has Km for S adenosylmethionine of 4.4 microM and specific activity 0.54 +/- 0.51 pmol/mg protein/15 min, is inhibited by S-adenosylhomocysteine, displays optimal activity at pH 8.0-9.5, and is stimulated by isoproterenol greater than epinephrine greater than norepinephrine, but not by prostaglandin E1, serum-treated zymosan, formyl-methionyl-leucyl-phenylalanine, or adenosine 3':5' cyclic monophosphate. Because activation of PEMT reportedly contributes to several processes known to be abnormal in cystic fibrosis, including coupling of the beta-adrenergic receptor to adenylate cyclase, activity of PEMT was compared in lymphocyte and granulocyte membrane preparations from cystic fibrosis patients and healthy controls, in which abnormal coupling of beta-adrenergic receptor to adenylate cyclase had been demonstrated. For both cell types, the Km and specific activity of PEMT were comparable in normal and cystic fibrosis samples. Therefore, the hypothesis that reduced PEMT activity accounts for the impaired coupling of beta adrenergic receptor to adenylate cyclase in lymphocytes and granulocytes in cystic fibrosis is rejected. PMID- 3025803 TI - Gluconeogenesis from lactate in the small intestinal mucosa of suckling rats. AB - Glucose formation from uniformly labeled 14C-lactate was studied in the small intestinal mucosa of rats and rabbits. It was found to occur in infant but not in adult (weaned) animals and to be increased by the presence of dibutyryl cyclic AMP or tetradecyl glycidic acid. Similarly the formation of glyceride glycerol was enhanced by tetradecyl glycidic acid but not by glucagon or cyclic AMP. The glycogen content of the intestinal mucosa was always low, but increased significantly at the time of weaning. It is suggested that gluconeogenesis occurs in the small intestinal mucosa of infant rodents to supply glucose to the muscular part of the small intestine. PMID- 3025805 TI - Lactoferrin inhibits prostaglandin E2 secretion by breast milk macrophages. AB - The interaction between human breast milk macrophages and lactoferrin (LF) in its native form was studied in in vitro culture. Competitive inhibition-binding studies with 125I-LF and unlabeled LF showed that a specific receptor for LF was present on breast milk macrophages. LF at concentrations of 10(-6) -10(-9)M resulted in a dose-dependent inhibition of prostaglandin E2 secretion by breast milk macrophages (control-45 +/- 7; LF 10(-6)M -9 +/- 1 ng/ml/10(6) cells). This inhibitory effect was also observed when the macrophages were stimulated with Concanavalin A (LF 10(-6) M -80 +/- 5; 10(-9) M -45 +/- 8% inhibition of prostaglandin E2 secretion by Concanavalin A stimulated macrophages). Lactalbumin and lactoglobulin had no effect. Similar concentrations of LF had no effect on lysozyme production. We also demonstrated that human milk macrophages are capable of eliciting an oxidative burst as measured by superoxide or hydrogen peroxide production when stimulated by phorbol myristate acetate in in vitro culture. (basal superoxide -1.4 +/- 0.3; phorbol myristate acetate 28.8 +/- 3.5 nmol/1 X 10(6) cells/90 min; basal hydrogen peroxide 11.7 +/- 4.6; phorbol myristate acetate -57.5 +/- 2.3 nmol/mg protein/90 min). LF had no effect on the oxidative burst. These results suggest that interaction of aqueous and cellular components of breast milk may occur and result in varied physiological effects. PMID- 3025804 TI - Effect of delta-9-tetrahydrocannabinol on the in vitro uptake of alpha-amino isobutyric acid by term human placental slices. AB - Tetrahydrocannabinol (THC), the active component in marijuana smoke, crosses the placenta and is a potential fetotoxic agent. In both human and animal studies, the most consistent fetal effect of THC is intrauterine growth retardation. Since fetal somatic growth is dependent on placental transfer of nutrients, including essential amino acids, we studied the effect of THC upon the in vitro uptake of amino acid by term human placental slices. Uptake of alpha-amino isobutyric acid was inhibited in a dose-dependent fashion, correlating with the log of the dose (1-100 microM THC; r = 0.945; p less than 0.01). Compared to control tissue, significant impairment of alpha-amino isobutyric acid uptake began at 20 microM THC. Similar results were found for valine. The time course (30-120 min) for alpha-amino isobutyric acid uptake showed linearity for both control and THC-(50 microM) treated tissue, but there was a marked reduction in the THC slope. Uptake of alpha-amino isobutyric acid was significantly reduced at all times. The sustained effect of THC was slightly, but significantly, reversed by removal of THC from the medium after 90 min of 50 microM THC exposure. Only partial reversal may have been due to the 15- to 20-fold accumulation of THC in the placental tissue. Uptake kinetics showed noncompetitive inhibition with decreased Vmax: control Vmax = 51.66 +/- 6.26 versus 50 microM THC = 26.96 +/- 6.22 (mmol/liter intracellular water per h) (p less than 0.01); and no change in diffusion constant (Km): control Km = 0.78 +/- 0.08 versus 50 microM THC = 0.80 +/- 0.09 (mM).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3025806 TI - Vesicoureteral reflux in asymptomatic siblings of patients with known reflux: radionuclide cystography. AB - The familial nature of vesicoureteral reflux among siblings of patients with vesicoureteral reflux has been reported to be from 8% to 32%. These included both symptomatic and asymptomatic siblings. The incidence of vesicoureteral reflux in asymptomatic siblings, however, has not been studied extensively. Sixty asymptomatic siblings of patients known to have vesicoureteral reflux were studied with radionuclide voiding cystography. Their ages ranged from 2 months to 15 years (mean, 4.2 years). Vesicoureteral reflux was detected in 27 of 60 (45%) of the siblings. Vesicoureteral reflux was unilateral in 15 and bilateral in 12 of the siblings. Radionuclide cystography is more sensitive than radiographic cystography and results in a very low radiation dose to the patient. The gonadal dose with radionuclide cystography is only 1.0 to 2.0 mrads. Because of these features, radionuclide cystography is a nearly ideal technique for the diagnosis of vesicoureteral reflux in siblings of patients with known vesicoureteral reflux. All siblings (symptomatic or asymptomatic) of patients with known vesicoureteral reflux should have a screening radionuclide cystography. PMID- 3025807 TI - Infant pneumonitis associated with cytomegalovirus, Chlamydia, Pneumocystis, and Ureaplasma: follow-up. AB - A total of 205 infants who were hospitalized when younger than 3 months of age for pneumonitis were followed longitudinally. Of these patients, 145 (70%) had evidence of infection with one or more pathogens. The most common etiologic agents were Chlamydia trachomatis 61/193 (36%), respiratory syncytial virus 33/142 (23%), cytomegalovirus 42/203 (20%), Pneumocystis carinii 30/171 (17%), and Ureaplasma urealyticum 21/125 (16%). The initial clinical presentation was characterized by cough, rales, normal temperature, and diffuse obstructive airways disease by chest roentgenogram. Regardless of etiology, significant association occurred for cough and cytomegalovirus, apnea and Pneumocystis, and conjunctivitis and Chlamydia. Longitudinal follow-up demonstrates a mortality of 7/205 (3.4%). Morbidity was manifest as recurrent wheezing episodes in 86/187 (46%) patients, persistently abnormal chest roentgenographic findings for at least 12 months in 17/109 (15%) patients, and abnormal pulmonary functions in 15/25 (60%) patients. These abnormalities occurred irrespective of prematurity, atopy, or the initial etiologic agent associated with the pneumonitis. These data add further evidence that respiratory infections during infancy may well be predecessors of obstructive airways disease in later life. PMID- 3025808 TI - The effect of cyanide on apparent potassium conductance across the peritubular cell membrane of frog proximal tubules. AB - To test for the effect of cyanide on frog proximal renal tubules the potential difference across the peritubular cell membrane (PDpt) has been recorded continuously before and during peritubular application of 1 mmol/l cyanide using conventional microelectrodes. Before application of cyanide PDpt amounts to -61.5 +/- 2.2 mV in the absence of luminal substrate. Cyanide depolarizes the peritubular cell membrane by +18.8 +/- 2.3 mV/10 min in the presence and by +4.5 +/- 0.9 mV/10 min in the absence of luminal substrate. The rapid depolarization of the cell membranes to addition of glucose to luminal perfusate is not significantly influenced by exposure to cyanide, whereas the influence of altered peritubular potassium concentration (from 3 to 9 mmol/l) is significantly reduced from +15.2 +/- 1.7 mV to +8.7 +/- 1.8 mV. Following exposure to cyanide the lumped resistance of the luminal and peritubular cell membranes increases significantly by 36 +/- 7%/6 min, and the cellular core resistance significantly by 14 +/- 6%/6 min. As a result, cyanide markedly decreases the peritubular potassium conductance, depolarizes the cell membranes and reduces the driving force for sodium coupled transport processes. Thus cyanide fully mimics the effects of ouabain, although cyanide in contrast to ouabain is expected to deplete the cells from ATP. In conclusion ATP/ADP is not likely to play a major role in the regulation of sodium coupled transport processes and peritubular potassium conductance in amphibian proximal tubules. PMID- 3025809 TI - Effect of steroid hormones on the regulation of uterine contractility. AB - The kinetics of myosin light chain (LC20) phosphorylation has been studied during in vitro contraction and relaxation of rat uteri. Phosphorylation preceded contraction and continued during tetanus induced by KCl; the degree of phosphorylation was proportional to the percentage of contraction. On the other hand, during relaxation induced by isoproterenol, the level of phosphorylation did not change in the seconds following relaxation. A rapid change in cAMP did not appear to trigger a rapid change in LC20 phosphorylation. In cyclic rats, progesterone decreased the extent of LC20 phosphorylation: 50%-60% of the LC20 was phosphorylated in untreated animals or estrogen treated animals. This value fell to 30% after progesterone treatment of cyclic rats. In ovariectomized rats, steroid hormones did not affect the phosphorylation reaction. Under the same conditions, the level of cAMP-dependent protein kinases did not change during the cycle, or after estradiol-treatment, but in cyclic rats it doubled after progesterone treatment. Hence, the decrease in the level of LC20 phosphorylation observed in cyclic rats treated by progesterone could be due to a decrease in myosin light chain kinase (MLCK) activity, resulting from a higher proportion of the phosphorylated form of this enzyme. The concomitant increase in the proportion of activatable cAMP-dependent protein kinases could favor the maintenance of a higher level of phosphorylated MLCK for longer periods of time. PMID- 3025810 TI - [Thickening of the posterior wall of the bronchus intermedius in patient with lung cancer]. PMID- 3025811 TI - [Diagnostic value of CT with hepatic arterial infusion of lipiodol (Lp-CT) for detection of daughter nodules of hepatocellular carcinoma--comparison with CT during arterial portography]. PMID- 3025813 TI - Xrep, a plasmid-stimulating X chromosomal sequence bearing similarities to the BK virus replication origin and viral enhancers. AB - The human X chromosome-linked fragment, "Xrep," was sequenced because it exerts a positive effect on plasmid growth in both E. coli and Saccharomyces cerevisiae. The sequence revealed three features similar to the human BK virus replication origin: Xrep has a true palindrome, CCTCC(T)3CCTCC, which is similar to "true" palindrome-like sequences found at the replication origins of polyoma [CCTC(T/C)10CTCC], BK [CCTC(A/G)8CCTCC] and SV40 [CCTCC(A)6GCCTCC] viruses. Twenty nucleotides away from the true palindrome, Xrep has the sequence GAATCCTATTCACTTTT while BK virus, the human analogue of SV40, has GAAATCCCTATTCTTTT in exactly the same position relative to the true palindrome. These two 17-mers differ only in the positions of two nucleotides comparing Xrep and BK virus. Also similar to the replication origins of DNA viruses, Xrep appears to have a cluster of enhancers adjacent to the origin-like sequences. Potent enhancer-like activity was detected in pSV1 X CAT/Xrep constructs. Xrep may originate from an endogenous virus, or from an X chromosomal replication origin. PMID- 3025812 TI - Comparison of promoter suppression in avian and murine retrovirus vectors. AB - Previously, we described "promoter suppression" in infectious retrovirus vectors with two genes and an internal promoter. Here, we examined several parameters of promoter suppression and found that the amount of suppression in an integrated retrovirus vector was dependent both on whether the vector was derived from spleen necrosis virus or murine leukemia virus and on which internal promoter was present in the vector. Murine leukemia virus vectors showed less suppression than analogous spleen necrosis virus vectors. Furthermore, the amount of suppression was not dependent on either the relative strengths of the promoters nor the distance between the promoters. Moreover, we found that in vectors in which one promoter was suppressed, there was an inverse correlation between the DNaseI sensitivity of the chromatin surrounding a promoter and the suppression of its expression. PMID- 3025814 TI - Characterization of a supercoil-dependent S1 sensitive site 5' to the Drosophila melanogaster hsp 26 gene. AB - We have analyzed the prominent supercoil-dependent S1 nuclease cleavage site 5' to hsp 26 in the plasmid 88B13, which contains 11.7 kilobases from the Drosophila locus 67B1. The double stranded cleavage product is generated by initial nicking on the purine strand, six preferred sites occurring between positions -96 and -90 (relative to the start of transcription) with weaker ones extending to position 84, followed by cleavage on the pyrimidine strand at positions -86 and -84. A derivative of 88B13, 88B13-X, was generated by insertion of an Xho I linker at position -84; this does not affect the positions or strand specificity of the S1 cleavage in that region. A small deletion, delta 41.1, removes the homopurine/homopyrimidine stretch from positions -86 to -132 and is no longer sensitive to cleavage by S1 nuclease 5' to hsp 26. Mung bean and P1 nucleases recognize the same site 5' to hsp 26 and give the same general pattern of cleavage. All three nucleases show an initial cleavage of 88B13 DNA at this site at pH 5.5 but not at pH 6.5, indicating that the DNA structure there may be pH dependent in vitro. PMID- 3025815 TI - Simian virus 40 DNA replication in vitro: purification and characterization of replication factors from mouse cells. AB - We have previously developed simian virus 40 (SV40) DNA replication system in vitro (Ariga and Sugano, J. Virol. 48, 481, 1983). This system is composed of human HeLa or mouse FM3A nuclear extract and cytoplasmic extract of SV40 infected CosI cells. Here FM3A nuclear extract was fractionated by DEAE Sephacel and single-stranded DNA cellulose chromatography into three components required for accurate in vitro SV40 DNA replication. One fraction (A fraction) contained DNA polymerase-primase, and the second component (B fraction) contained DNA topoisomerase. Third component was further purified to near homogenuity using DEAE-Sephacel, single-stranded DNA cellulose, and glycerol gradient centrifugation. The purified protein (named factor I) bound to the origin containing fragment of SV40 DNA. The factor I enhanced the initiation of SV40 DNA replication catalyzed by SV40 infected CosI cytoplasm alone. When all four fractions consisting of A, B fractions, factor I, and SV40 infected CosI cytoplasm were mixed together, the system was reconstituted, meaning that initiation and subsequent elongation were completed to generate the full sized daughter molecules. PMID- 3025816 TI - Kinetics of the proton-deuteron exchange at position H8 of adenine and guanine in DNA. AB - Proton-NMR has been used to determine the activation energies and pre-exponential factors for the deuterium exchange of AH8 in poly(dA-dT).poly(dA-dT), and for GH8 in poly(dG-dC).poly(dG-dC). No simple relationship between the kinetic parameters and molecular conformation was found. By addition of 4.5 M NaCl a transition from the B to the Z conformation was induced for poly(dG-dC).poly(dG-dC), and an increased exchange rate was observed. The exchange rate for poly(dA-dT).poly(dA dT) also increased below 64 degrees C, and a significant decrease in activation energy on addition of 4.5 M NaCl was observed. The exchange rates at T = 55.8 degrees C were also measured for the AH8 and GH8 in random sequence calf thymus DNA. From the difference in exchange rates, a method of preferential labeling of either the AH8 or the GH8 in high molecular weight DNA is evaluated. PMID- 3025817 TI - Nucleotide sequence of a retrotransposon 297 isolated from Drosophila simulans. PMID- 3025818 TI - lambda PJ4A, a lambda replacement vector carrying amber mutations for cloning of EcoRI fragments. PMID- 3025819 TI - Equilibrium radionuclide angiocardiography to select inotropic therapy in patients with left ventricular aneurysm. AB - Fourteen patients with postinfarctual ventricular aneurysm underwent equilibrium radionuclide angiocardiography at rest (ERNA) before and after oral digoxin administration in order to evaluate the effects of increasing myocardial contractility upon both ventricular aneurysm mechanical behaviour and global ventricular function. The ejection fraction (EF) was not significantly affected by digoxin therapy. However, digoxin induced changes in EF (delta EF) correlated inversely with changes in aneurysm size and directly with changes in the extent of the hypokinetic area. Two types of aneurysm were observed: high-compliance aneurysm the size of which increased after digoxin administration while both EF and the extent of the hypokinetic area fell, and low compliance aneurysm for which opposite changes occurred. This different behaviour of ventricular aneurysm may have important practical implications as surgery would be probably more effective than medical treatment in improving resting ventricular function in patients with high-compliance aneurysm. PMID- 3025820 TI - Survival of cytomegalovirus in paper diapers and saliva. AB - We studied the survival of human cytomegalovirus inoculated experimentally into saliva and urine-soaked paper diapers. Infectious virus could be recovered from saliva maintained at room temperature or 37 degrees C for up to 2 hours after inoculation. Cytomegalovirus survived in paper diapers for periods as long as 48 hours, with the quantity of virus in urine remaining relatively constant for the initial 12 hours. These observations provide further support for the concept that fomites may have a role in the transmission of cytomegalovirus infections. PMID- 3025821 TI - Failure of varicella-zoster immunoglobulin in modification of severe congenital varicella. PMID- 3025822 TI - Characterization of vasoactive intestinal peptide (VIP) receptors in mammalian lung. AB - 125I-VIP bound specifically to sites on human, rat, guinea pig, and rabbit lung membranes with a dissociation constant (KD) of 60-200 pM and binding site maxima of 200-800 fmol/mg of protein. The presence of a second lower affinity site was detected but not investigated further. High affinity 125I-VIP binding was reversible and displaced by structurally related peptides with an order of potency: VIP greater than rGRF greater than PHI greater than hGRF greater than secretin = Ac Tyr1 D Phe2 GRF. 125I-VIP has been covalently incorporated into lung membranes using disuccinimidyl suberate. Sodium dodecyl sulfate polyacrilamide gel electrophoresis of labeled human, rat, and rabbit lung membranes revealed major 125I-VIP-receptor complexes of: Mr = 65,000, 56,000, and 64,000 daltons, respectively. Guinea pig lung membranes exhibited two 125I-VIP receptor complexes of Mr = 66,000 and 60,000 daltons. This labeling pattern probably reflects the presence of differentially glycosylated forms of the same receptor since treatment with neuroaminidase resulted in a single homogeneous band (Mr = 57,000 daltons). Soluble covalently labeled VIP receptors from guinea pig and human lung bound to and were specifically eluted from agarose-linked wheat germ agglutinin columns. Our studies indicate that mammalian lung VIP receptors are glycoproteins containing terminal sialic acid residues. PMID- 3025823 TI - ACTH-immunoreactive neurons and their projections in the cat forebrain. AB - The organization of adrenocorticotropin (ACTH)-immunoreactive (IR) cell bodies and fibers in the cat forebrain is described. ACTH-IR cell bodies are found only in and around the arcuate nucleus of the hypothalamus (ARH). They are not detected elsewhere even after pretreatment with colchicine. ACTH-IR fibers are present in discrete areas of the hypothalamus, the septo-limbic areas and in the paraventricular thalamic nucleus. Complete electrolytic lesions of the ARH destroy ACTH-IR cell bodies as well as fibers in all parts of the brain. These results suggest that, in the cat forebrain, the ARH is the only source of ACTH-IR fibers. PMID- 3025824 TI - Neuropeptide Y (NPY) and the pig heart: release and coronary vasoconstrictor effects. AB - The effects of electrical stimulation of the stellate ganglia on the arterio venous concentration differences of neuropeptide Y (NPY)-like immunoreactivity (LI) over the pig heart were studied in vivo in relation to changes in heart rate and left ventricular pressure. Furthermore, the effects of NPY on coronary vascular tone were analysed in vivo and in vitro. Stellate ganglion stimulation at a high frequency (10 Hz) caused a clear-cut, long lasting increase in plasma levels of NPY-LI in the coronary sinus compared to the aorta, suggesting release of this peptide from sympathetic terminals within the heart. The stimulation evoked overflow of NPY-LI from the heart was enhanced about 3-fold by alpha adrenoceptor blockade using phenoxybenzamine, suggesting that NPY release is under prejunctional inhibitory control by noradrenaline (NA). Combined alpha- and beta-adrenoceptor blockade abolished most of the positive inotropic response of the heart upon stellate ganglion stimulation, while a considerable positive chronotropic effect remained. After guanethidine treatment, stellate ganglion stimulation still produced a small positive inotropic and chronotropic effect on the heart. The stimulation evoked NPY overflow was markedly reduced by guanethidine indicating an origin from sympathetic nerve terminals. Injection of NPY into the constantly perfused left anterior descending artery in vivo caused a long lasting, adrenoceptor antagonist resistant increase in perfusion pressure, suggesting coronary vasoconstriction. NPY contracted coronary arteries in vitro via a nifedipine-sensitive mechanism. NA dilated coronary vessels both in vivo and in vitro via beta-adrenoceptor activation. It is concluded that sympathetic nerve stimulation increases overflow of NPY-LI from the heart suggesting release from cardiac nerves in vivo.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3025825 TI - ACTH-(1-24) and alpha-MSH antagonize feeding behavior stimulated by kappa opiate agonists. AB - ACTH-(1-24) and alpha-MSH, intracerebroventricularly (ICV) injected at the doses of 4 and 10 micrograms/animal, respectively, markedly inhibited spontaneous feeding in adult Sprague-Dawley rats, the effect remaining significant for 6-9 hours. At these same doses, ACTH-(1-24) and alpha-MSH abolished the feeding stimulatory effect of the kappa opiate receptor agonist pentazocine, intraperitoneally (IP) injected at the dose of 10 mg/kg. The same antagonism was obtained by ICV injection of ACTH-(1-24) into rats IP treated with other kappa opiate agonists, bremazocine and tifluadom, at the doses of 1 and 5 mg/kg, respectively. These data suggest that melanocortin peptides play an inhibitory role in the complex regulation of food intake, and further support and extend the hypothesis of a melanocortin-opioid homeostatic system, its two neuropeptide components usually having opposite, mutually-balancing effects. PMID- 3025826 TI - A fragment of vasoactive intestinal peptide, VIP(10-28), is an antagonist of VIP in the colon carcinoma cell line, HT29. AB - The 19 amino acid carboxyl terminus fragment of vasoactive intestinal peptide (VIP), VIP(10-28), inhibits [125I]VIP binding in intact HT29 colonic adenocarcinoma cells and in membranes from these cells. However, VIP(10-28) alone has no effect on adenylate cyclase activity (membranes) or cyclic AMP synthesis (intact cells) in HT29 cells although VIP receptor agonists are markedly stimulatory. The indicated antagonist character of VIP(10-28) was confirmed by rightward parallel shifts of VIP dose response curves in the presence of VIP(10 28) in adenylate cyclase and cyclic AMP synthesis experiments. The equilibrium dissociation constant values for VIP(10-28) from these experiments agree with values from inhibition binding studies. The lack of effect of VIP(10-28) on forskolin dose response curves in HT29 adenylate cyclase assays indicates the specificity of the VIP(10-28) antagonism, thus suggesting that VIP(10-28) may be a useful tool in studying VIP receptor regulation and other aspects of the mechanisms of VIP action. The potential regulatory role of a proteolytically generated fragment of VIP acting antagonistically at VIP receptors is discussed. PMID- 3025827 TI - Binding sites of atrial natriuretic peptide in tree shrew adrenal gland. AB - Adrenal gland binding sites for atrial natriuretic peptide-(99-126) (ANP) were quantitated in tree shrew (Tupaia belangeri) by incubation of adrenal sections with (3-[125I]-iodotyrosyl28) atrial natriuretic peptide-(99-126), followed by autoradiography with computerized microdensitometry. In the adrenal glands, there are three types of ANP binding sites. One is located in the zona glomerulosa (BMax 84 +/- 6 fmol/mg protein; Kd 122 +/- 9 pM); the second in the zona fasciculata and reticularis (BMax 29 +/- 2 fmol/mg protein; Kd 153 +/- 6 pM) and the third in the adrenal medulla (BMax 179 +/- 1 fmol/mg protein; Kd 70 +/- 2 pM). Besides the influence of ANP on the regulation of adrenocortical mineralcorticoid and glucocorticoid secretion our findings raise the possibility for a local site of action of atrial natriuretic peptide in the regulation of adrenomedullary catecholamines in the tree shrew, primates and man. PMID- 3025828 TI - Naturally occurring opioid peptides modulate H+-production by isolated rat parietal cells. AB - The rationale for the present study was to determine the effects of naturally occurring opioid peptides on H+-production by isolated rat parietal cells as indirectly measured by [14C]-aminopyrine uptake. In crude preparations (18 to 25% parietal cells) and in enriched (80 to 90%) parietal cell fractions stimulation by submaximal histamine- or dibutyryl cAMP-concentrations (10(-6)-10(-4) mol/l) was augmented by 20-30% in the presence of methionine-enkephalin (Met-Enk) and Met-Enk Arg6Phe7 (10(-7) to 10(-5) mol/l). This augmentation was blocked by the opiate receptor antagonist (-)naloxone (10(-6) mol/l) suggesting specificity of the action of Met-Enk and Met-Enk Arg6Phe7. At 10(-6) mol/l (-)naloxone did not exert nonspecific toxic effects. Yet, even in the absence of exogenous opioids, histamine-induced H+-production was inhibited by 3 X 10(-5) or 10(-4) mol/l ( )naloxone. Since similar inhibition occurred with (+)naloxone, an inactive stereoisomer which does not interact with opiate receptors, effects of ( )naloxone at concentrations above 10(-5) mol/l must be considered nonspecific. We conclude that Met-Enk and Met-Enk Arg6Phe7 have no effect on basal, but augment stimulated H+-production by a direct effect on the parietal cells. At nontoxic concentrations (-)naloxone antagonizes this augmentation indicating that it is mediated by specific opiate receptors on the parietal cells. PMID- 3025829 TI - Enzymatic activity of glycogen metabolism in chorionic villi. AB - Glycogen content and six major enzymatic activities involved in glycogen metabolism were analysed in chorionic villi (CV). Glycogen levels were found to be lower than those known to exist in liver and muscle. Activities of alpha glucosidase, amylo-1,6-glucosidase, phosphorylase b and phosphorylase kinase were detectable by standard methods. The enzymatic activities of glucose-6-phosphatase and phosphorylase a were undetectable. These findings suggest that CV biopsies can be useful for first-trimester diagnosis of glycogen storage disease types II, III and VI, but not for type I (glucose-6-phosphatase deficiency). PMID- 3025830 TI - [Importance of Foscarnet in the treatment of cytomegalovirus pneumopathy in a patient with acquired immunodeficiency syndrome]. PMID- 3025831 TI - An Epstein-Barr virus transcript from a latently infected, growth-transformed B cell line encodes a highly repetitive polypeptide. AB - By screening a cDNA library prepared from poly(A)+ RNA isolated from an Epstein Barr virus latently infected, growth-transformed human B-lymphoblastoid cell line, we have recovered a clone corresponding to a highly spliced viral transcript encoded largely by the major internal repeat (IR1). The 5' region contains one copy of a 26-base-pair (bp) exon (W0) [which is 28 bp downstream from a CAATT-(N)34TATAAA sequence (N, unspecified base)] and seven copies of two small exons (W1, 66 bp; W2, 132 bp). In addition, there are three exons from the "unique" region of the BamHI Y fragment of the viral genome. Two other cDNA clones that have been described, corresponding to latent viral transcripts, share homology in their 5' regions with this clone and are clearly divergent at their 3' ends. The cDNA clone described in this paper contains one long open reading frame that extends through the repeat element. In vitro transcription and translation of this open reading frame yielded a 62-kDa polypeptide that could be immunoprecipitated by an Epstein-Barr virus-positive human serum. PMID- 3025832 TI - Cloning and characterization of the high-affinity cAMP phosphodiesterase of Saccharomyces cerevisiae. AB - A gene, PDE2, has been cloned from the yeast Saccharomyces cerevisiae that, when present in high copy, reverses the phenotypic effects of RAS2Val19, a mutant form of the RAS2 gene that renders yeast cells sensitive to heat shock and starvation. It has previously been shown that the RAS proteins are potent activators of yeast adenylate cyclase. We report here that PDE2 encodes a high-affinity cAMP phosphodiesterase that shares sequence homology with animal cell phosphodiesterases. These results therefore imply that the effects of RAS2Val19 are mediated through its changes in cAMP concentration. PMID- 3025833 TI - Identification of a conserved domain among cyclic nucleotide phosphodiesterases from diverse species. AB - Partial amino acid sequences have been determined for the Ca2+/calmodulin stimulated cyclic nucleotide phosphodiesterase from bovine brain and the cGMP stimulated cyclic nucleotide phosphodiesterase from bovine heart. Examination of these sequences for homologous segments and comparison with protein sequences derived from the nucleotide sequences of the yeast PDE2 gene and the Drosophila dunce+ gene [Chen, C.-N., Denome, S. & Davis, R. L. (1986) Proc. Natl. Acad. Sci. USA 83, 9313-9317; Sass, P., Field, J., Nikawa, J., Toda, T. & Wigler, M. (1986) Proc. Natl. Acad. Sci. USA 83, 9303-9307] reveal a 200- to 270-residue segment in each that is homologous to the others. The molecular masses of the four proteins vary from 40 kDa to 105 kDa, and the structural resemblance appears to be constrained to a single segment of each protein. These related segments are proposed to comprise the catalytic domains in this set of enzymes. The lack of absolute sequence identity between the two bovine enzymes shows that they are unique gene products that are not produced by alternative processing of a larger protein or of a single mRNA precursor. The data also strongly support the conclusion that the dunce+ gene locus of Drosophila and the PDE2 gene locus in yeast code for structural genes of cyclic nucleotide phosphodiesterases. PMID- 3025835 TI - Polarized expression of a chimeric protein in which the transmembrane and cytoplasmic domains of the influenza virus hemagglutinin have been replaced by those of the vesicular stomatitis virus G protein. AB - In polarized epithelial cells, influenza virus buds exclusively from the apical domain of the plasma membrane, whereas vesicular stomatitis virus (VSV) buds exclusively from the basolateral domain. In virus-infected cells, the envelope proteins, influenza hemagglutinin (HA) and vesicular stomatitis virus G (VSV G), are likewise transported to and localized in the same domain of the plasma membrane from which the viruses bud. Previous studies have shown that influenza HA and VSV G proteins, when expressed from cloned cDNAs, are accumulated preferentially on the proper domains (apical and basolateral, respectively), indicating that the signal(s) for polarized transport resides in the polypeptide backbone of the proteins. To further elucidate the structural features required for apical vs. basolateral transport, we have constructed a gene that encodes a chimeric protein (H1GA) containing the external domain of HA and the transmembrane and cytoplasmic domains of VSV G. When the chimeric protein (H1GA) is expressed in CV1 cells using a simian virus 40 late expression vector, it is transported to the cell surface with kinetics similar to that of the native HA protein. Further, the chimeric protein, when expressed in polarized MDCK cells using a vaccinia virus early expression vector, is transported only to the apical surface, suggesting that the ectodomain of HA contains a signal for apical transport. PMID- 3025834 TI - Molecular analysis of cDNA clones and the corresponding genomic coding sequences of the Drosophila dunce+ gene, the structural gene for cAMP phosphodiesterase. AB - We have isolated and sequenced cDNA clones representing portions of the polyadenylylated transcripts of the dunce+ gene. These define an open reading frame of 1086 bases and some of the 5'- and 3'-untranslated regions of the transcripts. The deduced amino acid sequence is strikingly homologous to the amino acid sequence of a Ca2+/calmodulin-dependent cyclic nucleotide phosphodiesterase isolated from bovine brain and more weakly related to the predicted amino acid sequence of a yeast cAMP phosphodiesterase. These homologies, together with prior genetic and biochemical studies, provide unambiguous evidence that dunce+ codes for a phosphodiesterase. In addition, the dunce+ gene product shares a seven-amino acid sequence with a regulatory subunit of cAMP-dependent protein kinase that is predicted to be part of the cAMP binding site. We also identify a weak homology between a region of the dunce+ gene product and the egg-laying hormone precursor of Aplysia californica. The open reading frame is divided in the genome by four introns. PMID- 3025836 TI - Angiotensin-stimulated production of inositol trisphosphate isomers and rapid metabolism through inositol 4-monophosphate in adrenal glomerulosa cells. AB - The production and metabolism of inositol phosphates in rat adrenal glomerulosa cells prelabeled with [3H]inositol and stimulated with angiotensin II were analyzed by high-performance anion-exchange chromatography. Exposure to angiotensin II was accompanied by a rapid and substantial decrease in the phospholipid precursor, phosphatidylinositol (PtdIns) 4,5-bisphosphate with only a slight and transient increase in the level of the biologically active product, inositol 1,4,5-trisphosphate (Ins-1,4,5-P3), to a peak at about 5 sec. Inositol 1,3,4-trisphosphate (Ins-1,3,4-P3), the putative metabolite of Ins-1,4,5-P3, was also formed rapidly and maintained an elevated steady-state level during stimulation by angiotensin II. Inositol 1,4-bisphosphate (Ins-1,4-P2) exhibited a simultaneous and prominent increase that could not be accounted for solely by direct breakdown of PtdIns 4-phosphate, indicating that large amounts of Ins 1,4,5-P3 must also have been produced and metabolized. The rapid formation of a substantial amount of inositol 4-monophosphate (Ins-4-P), with no significant change in the level of inositol 1-monophosphate (Ins-1-P) during the first minute of stimulation, was a notable feature of the glomerulosa cell response to angiotensin II. These observations indicate that PtdIns-4,5-P2 catabolism in the angiotensin-stimulated glomerulosa cell initially proceeds via Ins-1,4,5-P3 through Ins-1,3,4-P3 and Ins-1,4-P2 to form Ins-4-P rather than Ins-1-P and that direct hydrolysis of PtdIns by phospholipase C does not occur during the initial phase of angiotensin action. In glomerulosa cells stimulated by angiotensin II in the presence of Li+, the progressive accumulation of both Ins-4-P, and after a short lag period, Ins-1-P indicated that dephosphorylation of both isomers was inhibited by Li+. The increase of Ins-P isomers in the presence of Li+ was associated with increased and progressive accumulation of Ins-1,4-P2 and Ins 1,3,4-P3 but not of Ins-1,4,5-P3. These data demonstrate that sustained and massive breakdown of PtdIns phosphates begins within seconds during cell activation by angiotensin II. The Ca2+-mobilizing metabolite, Ins-1,4,5-P3, is rapidly converted to Ins-1,3,4-P3 and degraded through Ins-1,4-P2 and Ins-4-P, in contrast to the previous view that conversion to Ins-1-P is the major route of PtdIns 4,5-bisphosphate metabolism. PMID- 3025837 TI - Photoaffinity labeling of the porcine brain alpha 2-adrenergic receptor using a radioiodinated arylazide derivative of rauwolscine: identification of the hormone binding subunit. AB - A functionalized derivative of the alpha 2-selective antagonist rauwolscine formed the basis for a photoaffinity adduct that has allowed identification of the hormone-binding subunit of the brain alpha 2-adrenergic receptor protein. Rauwolscine carboxylate underwent reaction with 4-N-t-butyloxycarbonyl aminoaniline, leading to the synthesis of rauwolscine 4-aminophenyl carboxamide (Rau-AmPC). Rau-AmPC was radioiodinated and converted to the arylazide derivative, 17 alpha-hydroxy-20 alpha-yohimban-16 beta-[N-(4-azido-3 [125I]iodo)phenyl] carboxamide (125I-Rau-AzPC), via a diazonium salt intermediate. The characterization of 125I-Rau-AzPC as a photolabile probe employed alpha 2-adrenergic receptors, which were first solubilized from porcine brain membranes and partially purified by affinity chromatography utilizing a yohimbine-agarose affinity matrix. In the partially purified receptor preparation incubated with 125I-Rau-AzPC, photolysis resulted in covalent labeling of a major (Mr, 62,000) peptide as determined by NaDodSO4/PAGE and autoradiography. Labeling of this peptide was inhibited by the alpha 2-selective antagonist, yohimbine, and the non-subtype-selective alpha-antagonist, phentolamine, but not by the alpha 1 antagonist, prazosin, or the beta-receptor antagonist, (-)-alprenolol. The alpha adrenergic agonist epinephrine also inhibited labeling in a stereoselective manner. These data indicate that the photolabeled Mr 62,000 peptide is the hormone-binding subunit of the alpha 2-adrenergic receptor protein. The availability of this radioiodinated photoaffinity probe for the alpha 2 adrenergic receptor should facilitate further structural and biophysical characterization of the receptor protein. PMID- 3025838 TI - Two DNA recognition domains of the specificity polypeptides of a family of type I restriction enzymes. AB - The hsd genes of Salmonella typhimurium and Salmonella potsdam encode related type I restriction and modification systems designated SB and SP, respectively; the polypeptide encoded by the hsdS gene dictates the DNA sequence recognized. The hsdS genes of the SB and SP systems have a conserved sequence of around 100 base pairs flanked by two nonhomologous (variable) regions of around 500 base pairs. Recombination between the hsdS genes of SB and SP generated a system (SQ) with a different recognition specificity. We have localized the position of the crossover in the central conserved region by analysis of nucleotide sequences. Concomitant with the generation of a new combination of flanking variable regions is the recombination of minor differences in the central conserved region. A polypeptide domain encoded on the 5' side of the crossover dictates recognition of the trinucleotide component of the target sequence, and a second domain, encoded on the 3' side of the crossover, similarly governs recognition of the tetra- or penta-nucleotide component. Our analysis implicates at least parts of the variable regions in the determination of the specificity of interaction between protein and DNA. Furthermore, the trinucleotide components of the recognition sequences of S. typhimurium and Escherichia coli K-12 are identical, and the 5' segments of their hsdS genes are strikingly homologous rather than variable. PMID- 3025840 TI - Functional expression of a human Na+/H+ antiporter gene transfected into antiporter-deficient mouse L cells. AB - To clone the gene for the human Na+/H+ antiporter, we first constructed a stable mouse LTK- cell line (LAP1) lacking Na+/H+ antiport activity. Second, we devised a selective technique based on acid killing that specifically sorts out cells expressing low levels of Na+/H+ antiport activity from a population of antiporter deficient cells (AP-). LAP1 cells (TK- and AP-) were cotransformed with human genomic DNA and the thymidine kinase (TK) gene. TK+ transformants, first selected, were submitted to acid loading. The rare transformants that survived (frequency, 2-8 X 10(-7) expressed Na+/H+ antiport activity (AP+). We found that: transformation with mouse LAP1 DNA did not give rise to AP+ transformants; transformation of LAP1 cells with DNA from an altered Na+/H+ antiporter hamster variant led to AP+ transformants expressing the altered Na+/H+ antiporter of the DNA donor; human repeated sequences were present in all primary, secondary, and tertiary mouse AP+ transformants; six identical EcoRI human DNA fragments (55 kilobase pairs of the human genome) cosegregated with the Na+/H+ antiport activity in secondary and tertiary transformants. These results strongly suggest that we have stably expressed the structural gene for the human Na+/H+ antiporter in mouse cells. PMID- 3025839 TI - Diacylglycerol-induced translocation of diacylglycerol kinase: use of affinity purified enzyme in a reconstitution system. AB - Diacylglycerol-induced translocation of diacylglycerol kinase (ATP:1,2 diacylglycerol 3-phosphotransferase, EC 2.7.1.107) from the soluble to the membrane-bound compartments was demonstrated both in crude tissue homogenates and in a reconstituted enzyme-membrane model system. In homogenates of either rat brain or liver, incubation with diacylglycerol or phospholipase C, but not phospholipase A2 or phospholipase D, resulted in the translocation of diacylglycerol kinase activity from the soluble to the particulate fraction. This observation formed the basis for the first step in a two-step purification of diacylglycerol kinase. Enzyme extracted in 1 M salt from membranes of rat brain homogenates made in the presence of phospholipase C was purified further by affinity chromatography on a column containing phosphatidylserine, diacylglycerol, and cholesterol immobilized in polyacrylamide. This step yielded an enzyme preparation (step 2 enzyme) that was 500- to 750-fold purified (relative to the tissue homogenate) and required phosphatidylserine for stability. All other lipids tested failed to stabilize the enzyme. The properties of the enzyme preparation were similar to those of mammalian diacylglycerol kinases described by others. Reconstitution experiments showed that the soluble step 2 enzyme bound to inside-out vesicles of human erythrocytes only in the presence of diacylglycerol or phospholipase C but not phospholipase A2 or D. Redistribution of the kinase from soluble to vesicle-bound forms occurred rapidly and was dependent on the concentration of phospholipase C used to treat the vesicles. Physiological concentrations of calcium (50-1000 nM) did not enhance the phospholipase C-mediated translocation of the kinase. Thus, diacylglycerol kinase can translocate from cytosol to membranes in a manner dependent on the content of membrane-bound diacylglycerol but independent of the ambient concentration of calcium. PMID- 3025842 TI - Phosphatidylinositol from ethanol-fed rats confers membrane tolerance to ethanol. AB - The presence of ethanol disorders (fluidizes) biological membranes, but its chronic administration confers resistance to this perturbation (membrane tolerance). The latter effect has been invoked as an explanation for behavioral tolerance in alcoholics, but the molecular basis for membrane tolerance is obscure. To study the molecular mechanisms of this acquired resistance to disordering, we fed rats ethanol (36% of total calories) for 35 days, after which we quantitatively separated the phospholipids of hepatic microsomal membranes by high-performance liquid chromatography. Multilamellar vesicles were prepared from the recombined phospholipid classes, and their physical properties were examined by electron spin resonance. Vesicles composed of phospholipids from untreated rats were disordered (fluidized) in the presence of ethanol, whereas those made from phospholipids of ethanol-fed rats were resistant to this effect. When phosphatidylcholine (66.5 mol %), phosphatidylethanolamine (21 mol %), or phosphatidylserine (4.0 mol %) from ethanol-fed rats replaced their corresponding phospholipids in vesicles prepared from microsomal phospholipids from untreated rats, the membranes were still disordered by ethanol. In contrast, when 2.5-8.5 mol % phosphatidylinositol from ethanol-fed rats replaced phosphatidylinositol from untreated rats, the reconstituted membranes were rendered resistant to ethanol-induced disordering. Liver microsomal phosphatidylinositol (2.5-8.5 mol %) from ethanol-fed rats also conferred membrane tolerance to vesicles composed of bovine liver and brain phospholipids, an effect which demonstrates that the ability of phosphatidylinositol to confer membrane tolerance is not restricted to the microsomal membrane. PMID- 3025841 TI - Structural analysis of a rat liver glutathione S-transferase Ya gene. AB - We have isolated and characterized a complete structural gene encoding a rat liver glutathione S-transferase (glutathione transferase; EC 2.5.1.18) Ya subunit. The gene spans approximately 11 kilobases and is comprised of seven exons separated by six introns. A sequence similar to the Goldberg-Hogness promoter ("TATA" box), TATTA, is located 32 base pairs upstream from the transcription initiation site. Exons 2 and 4 of the glutathione S-transferase gene encode amino acid sequences of the Ya subunit that are highly conserved in the Yc subunit, whereas exons 3 and 5 encode amino acids that are divergent in the Yc subunit. These data suggest that exons 2 and 4 may encode domains of the Ya subunits that have similar structural or functional properties to the corresponding domains in the Yc subunit (e.g., glutathione binding site), whereas exons 3 and 5 may encode domains of the Ya subunit that have unique structural or functional properties to the corresponding domains in the Yc subunit (e.g., substrate binding site). PMID- 3025843 TI - Phosphorylation/dephosphorylation of the beta-adrenergic receptor regulates its functional coupling to adenylate cyclase and subcellular distribution. AB - Prolonged exposure of cells or tissues to drugs or hormones such as catecholamines leads to a state of refractoriness to further stimulation by that agent, known as homologous desensitization. In the case of the beta-adrenergic receptor coupled to adenylate cyclase, this process has been shown to be intimately associated with the sequestration of the receptors from the cell surface through a cAMP-independent process. Recently, we have shown that homologous desensitization in the frog erythrocyte model system is also associated with increased phosphorylation of the beta-adrenergic receptor. We now provide evidence that the phosphorylation state of the beta-adrenergic receptor regulates its functional coupling to adenylate cyclase, subcellular translocation, and recycling to the cell surface during the process of agonist induced homologous desensitization. Moreover, we show that the receptor phosphorylation is reversed by a phosphatase specifically associated with the sequestered subcellular compartment. At 23 degrees C, the time courses of beta adrenergic receptor phosphorylation, sequestration, and adenylate cyclase desensitization are identical, occurring without a lag, exhibiting a t1/2 of 30 min, and reaching a maximum at approximately 3 hr. Upon cell lysis, the sequestered beta-adrenergic receptors can be partially recovered in a light membrane vesicle fraction that is separable from the plasma membranes by differential centrifugation. The increased beta-adrenergic receptor phosphorylation is apparently reversed in the sequestered vesicle fraction as the sequestered receptors exhibit a phosphate/receptor stoichiometry that is similar to that observed under basal conditions. High levels of a beta-adrenergic receptor phosphatase activity appear to be associated with the sequestered vesicle membranes. The functional activity of the phosphorylated beta-adrenergic receptor was examined by reconstituting purified receptor with its biochemical effector the guanine nucleotide regulatory protein (Ns) in phospholipid vesicles and assessing the receptor-stimulated GTPase activity of Ns. Compared to controls, phosphorylated beta-adrenergic receptors, purified from desensitized cells, were less efficacious in activating the Ns GTPase activity. These results suggest that phosphorylation of the beta-adrenergic receptor leads to its functional uncoupling and physical translocation away from the cell surface into a sequestered membrane domain. In the sequestered compartment, the phosphorylation is reversed thus enabling the receptor to recycle back to the cell surface and recouple with adenylate cyclase. PMID- 3025844 TI - Actin-severing activity copurifies with phosphofructokinase. AB - Microinjection of muscle 6-phosphofructokinase (PFK; EC 2.7.1.11) into tissue culture cells led to a reversible disintegration of microfilament bundles (stress fibers). The mode of disruption as well as of recovery of stress fibers was very similar to that found previously in experiments performed with the actin-severing protein brevin, an extracellular variant of gelsolin. PFK, like brevin, was also capable of disrupting stress fibers in detergent-extracted cells and in ethanol fixed cells, in a Ca2+-dependent manner. When compared with heart muscle gelsolin, PFK comigrated with the 85- to 90-kDa band. Antibodies against PFK crossreacted with gelsolin from the same species. These results point to a tight association between polypeptides with similar biochemical and immunological parameters present in both preparations. They suggest hitherto unexpected cellular control mechanisms for both microfilament functions and glycolysis. PMID- 3025845 TI - 2-(N-acetoxy-N-acetylamino)fluorene mutagenesis in mammalian cells: sequence specific hot spot. AB - Mutations induced by 2-(N-acetoxy-N-acetylamino)fluorene were studied using temperature-sensitive simian virus 40 (SV40) mutants as probe in monkey kidney cells. In vitro treatment of the SV40 virions with 2-(N-acetoxy-N acetylamino)fluorene increased mutagenesis and decreased survival in the viral progeny. A lethal hit of approximately 85 acetylaminofluorene adducts per SV40 genome was calculated. UV irradiation of cells prior to infection did not modify the results. Molecular analysis of independent SV40 revertants showed that 2-(N acetoxy-N-acetylamino)fluorene induces base substitutions that are located not opposite putative acetylaminofluorene adducts but next to them. Moreover, a hot spot of mutation restoring a true wild-type genotype was observed in 10 of the 16 revertants analyzed. This hot spot, not targeted opposite a major DNA lesion, was not observed using UV light as damaging agent in the same genetic assay. Two models involving the stabilization, by acetylaminofluorene adducts, of the secondary structure of a specific quasipalindromic SV40 sequence are proposed to explain this sequence-specific hot spot. PMID- 3025846 TI - Molecular cloning, encoding sequence, and expression of vaccinia virus nucleic acid-dependent nucleoside triphosphatase gene. AB - A rabbit poxvirus genomic library contained within the expression vector lambda gt11 was screened with polyclonal antiserum prepared against vaccinia virus nucleic acid-dependent nucleoside triphosphatase (NTPase)-I enzyme. Five positive phage clones containing from 0.72- to 2.5-kilobase-pair (kbp) inserts expressed a beta-galactosidase fusion protein that was reactive by immunoblotting with the NTPase-I antibody. Hybridization analysis allowed the location of this gene within the vaccinia HindIIID restriction fragment. From the known nucleotide sequence of the 16-kbp vaccinia HindIIID fragment, we identified a region that contains a 1896-base open reading frame coding for a 631-amino acid protein. Analysis of the complete sequence revealed a highly basic protein, with hydrophilic COOH and NH2 termini, various hydrophobic domains, and no significant homology to other known proteins. Translational studies demonstrate that NTPase-I belongs to a late class of viral genes. This protein is highly conserved among Orthopoxviruses. PMID- 3025847 TI - Molecular analysis of the maize anthocyanin regulatory locus C1. AB - The C1 gene of maize plays a regulatory role in the production of anthocyanin pigments in the aleurone layer of the endosperm. As an initial step toward understanding the molecular details of how C1 controls pigment biosynthesis, we cloned the C1 gene. This was accomplished by first cloning a mutable allele of C1, c1-m5, which contains the transposable element Spm. A combination of molecular and genetic analysis was used to identify the Spm at the C1 locus. Individual genomic DNAs from a population in which the c1-mutable phenotype was segregating with the recessive c1 phenotype were digested with methyl-sensitive restriction enzymes and probed with a small DNA fragment derived from a defective Spm. One Sal I restriction fragment complementary to the Spm probe was shown to be present in the DNA of individuals with the c1-m5 phenotype but absent from DNA of individuals with a recessive c1 phenotype. Subsequent cloning and restriction analysis of this fragment revealed sequences flanking the Spm that proved to be C1-specific. A DNA fragment derived from the flanking sequences was then used as a probe to clone the wild-type C1 gene and several additional alleles of C1, including one stable recessive, two mutations caused by Ds insertions, one mutation induced by insertion of a defective Spm, and two dominant mutations, C1 S and C1-I. RNA blot hybridization analysis of three C1 alleles indicates that C1 regulation of the Bz1 and A1 structural genes in the anthocyanin biosynthetic pathway is at the transcriptional level. PMID- 3025848 TI - Elevated levels of mRNA can account for the trans-activation of human immunodeficiency virus. AB - The genome of human immunodeficiency virus encodes a protein that dramatically elevates amounts of viral proteins. The precise mechanism of this trans activation remains to be established. It has been reported that trans-activation can occur without major changes in the levels of mRNA. We constructed recombinant plasmids containing those viral sequences required in cis for trans-activation linked to the chloramphenicol acetyltransferase gene. These plasmids were introduced into cultured cells in either the presence or absence of a second plasmid that directed expression of the viral trans-activator protein. Expression of the chloramphenicol acetyltransferase gene was measured at the level of protein (by enzymatic assay) and RNA (by ribonuclease protection and primer extension). Our results demonstrate that trans-activation is accompanied by large increases in mRNA levels; these increases may be sufficient to explain the elevated levels of trans-activated protein. PMID- 3025849 TI - Blockade of electrical activity promotes the death of mammalian retinal ganglion cells in culture. AB - During the first 2 postnatal weeks, up to 50% of the ganglion cells in the mammalian retina normally die. Natural cell death may result from several factors, and electrical activity has been proposed as one critical element. Recent experiments in vivo using intraocular injection of tetrodotoxin (TTX) have suggested that competition for survival between ganglion cells from the two eyes is mediated by their degree of neuronal activity. In addition, the level of activity of afferents to the ganglion cells has been postulated to be an important variable in determining their survival. To investigate the mechanism of cell death engendered by altered activity, I studied the effect of electrical blockade with TTX (to block sodium channels and thus action potentials) or low Ca/high Mg (to block transmitter release and hence synaptic activity) on individual neurons in vitro. For this purpose, identified retinal ganglion cells (RGCs) from postnatal rats were maintained in culture. Unlike the previous in vivo experiments, this approach permitted the exact concentration of each agent to be controlled and the electrical activity of the RGCs to be recorded. In cultures from animals of postnatal day 2-10 (P2-10), 1 microM TTX or 0.2 mM Ca/20 mM Mg resulted in the death of about 50% of the RGCs, representing those cells that had displayed spontaneous electrical activity, but did not affect RGCs that lacked activity. However, the death of RGCs with spontaneous activity from P11-13 animals was not influenced by these drugs. These findings suggest that during a critical period of development neurons become dependent upon electrical activity, and the cessation of this activity can result in their death. In addition, conditioned medium, collected from cultures lacking TTX, rescued from death a large proportion of TTX-treated RGCs. Thus, the critical element for survival may represent modulation of a trophic factor related to the level of activity rather than electrical activity itself. Since, in vivo, natural cell death occurs in neurons of similar type and age, and in the same proportion as that induced by the artificial blockade of electrical activity in culture, these findings may be germane to the mechanism of natural cell death in the retina. PMID- 3025850 TI - Mutational analysis of the signal-anchor domain of influenza virus neuraminidase. AB - Influenza virus neuraminidase (NA; EC 3.2.1.18) possesses a signal-anchor hydrophobic domain at the amino terminus. To characterize the nature of this signal-anchor domain we have introduced single amino acid changes in this domain by oligonucleotide-directed mutagenesis. Three mutant NA proteins that were synthesized contained a single charged amino acid residue in place of a hydrophobic amino acid residue at position 11, 17, or 26 of the signal-anchor domain. When the altered NA proteins were expressed in CV-1 cells, two phenotypes were observed: substitution of arginine in place of glycine at position 11 and substitution of aspartic acid for valine at position 17 did not abolish the signal, the anchor, or the transport functions. On the other hand, substitution of arginine for isoleucine at position 26 blocked the migration of the NA protein from the Golgi complex to the cell surface. Double mutants were constructed from these single point mutations and they exhibited two phenotypes: one double mutant (aspartic acid at position 17 and arginine at position 26) was present mostly in the cytoplasm and the other (arginine at positions 11 and 26) was present mostly in the rough endoplasmic reticulum. These results indicate that the hydrophobic amino acids at positions 11, 17, and 26 are required for intracellular transport. Furthermore, the accumulation of the mutant proteins in the rough endoplasmic reticulum or the Golgi apparatus suggests the existence of putative intracellular transport (or traffic) signals in the signal-anchor domain of NA. PMID- 3025852 TI - Asymmetrical pairings of transposons in and proximal to the white locus of Drosophila account for four classes of regularly occurring exchange products. AB - An explanation for the origins of four classes of regularly occurring duplication and deficiency chromosomes is provided through examination of their molecular structures. The duplications and deficiencies occur as the reciprocal products of crossing-over, following two different patterns of asymmetrical synapsis between transposons positioned in and proximal to the white locus of Drosophila melanogaster. Three copies of the retrovirus-like transposon roo are involved in the exchanges. Evidence suggests that transposon-mediated asymmetrical exchange is a general phenomenon in eukaryotes, which adds significantly to the effects of transposons in the restructuring of eukaryotic genomes. PMID- 3025851 TI - Simian virus 40 (SV40) DNA replication: SV40 large T antigen unwinds DNA containing the SV40 origin of replication. AB - The simian virus 40 (SV40) large T antigen (large tumor antigen), in conjunction with a topoisomerase, a DNA binding protein, and ATP, catalyzed the conversion of a circular duplex DNA molecule containing the SV40 origin of replication to a form with unusual electrophoretic mobility that we have named form U. Analysis of this molecule revealed it to be a highly underwound covalently closed circle. DNA unwinding was not detected with DNA containing a SV40 T-antigen binding site II mutation that renders the DNA inactive in replication. The unwinding reaction requires the action of a helicase, and SV40 T-antigen preparations contain such an activity. The T-antigen-associated ability to unwind DNA copurified with other activities intrinsic to T antigen [ability to support replication of SV40 DNA containing the SV40 origin, poly(dT)-stimulated ATPase activity, and DNA helicase]. However, in contrast to the unwinding activity, the SV40 T-antigen associated helicase activity was not sequence-specific. A variety of labeled oligonucleotides hybridized with circular single-stranded DNA were displaced by T antigen in the presence of ATP. PMID- 3025853 TI - Nucleotide sequence of a preferred maize chloroplast genome template for in vitro DNA synthesis. AB - Maize chloroplast DNA sequences representing 94% of the chromosome have been surveyed for their activity as autonomously replicating sequences in yeast and as templates for DNA synthesis in vitro by a partially purified chloroplast DNA polymerase. A maize chloroplast DNA region extending over about 9 kilobase pairs is especially active as a template for the DNA synthesis reaction. Fragments from within this region are much more active than DNA from elsewhere in the chromosome and 50- to 100-fold more active than DNA of the cloning vector pBR322. The smallest of the strongly active subfragments that we have studied, the 1368-base pair EcoRI fragment x, has been sequenced and found to contain the coding region of chloroplast ribosomal protein L16. EcoRI fragment x shows sequence homology with a portion of the Chlamydomonas reinhardtii chloroplast chromosome that forms a displacement loop [Wang, X.-M., Chang, C.H., Waddell, J. & Wu, M. (1984) Nucleic Acids Res. 12, 3857-3872]. Maize chloroplast DNA fragments that permit autonomous replication of DNA in yeast are not active as templates for DNA synthesis in the in vitro assay. The template active region we have identified may represent one of the origins of replication of maize chloroplast DNA. PMID- 3025854 TI - Phorbol ester-induced growth arrest of murine myelomonocytic leukemic cells with virus-disrupted myb locus is not accompanied by decreased myc and myb expression. AB - A class of mouse neoplasms termed Abelson virus-induced plasmacytoid lymphosarcomas has previously been found to express abnormal myb transcripts due to a helper virus insertion into the 5' end of the c-myb locus. We have established clonal cell lines from these tumors and have shown that they all express markers characteristic of myelomonocytic, rather than lymphoid, cells. Treatment of the plasmacytoid lymphosarcoma lines with phorbol 12-myristate 13 acetate led to various degrees of growth arrest, presumably due to myelomonocytic differentiation. To date, characterization of myeloid cell lines with varying responses to phorbol 12-myristate 13-acetate has not been reported. The different clonal plasmacytoid lymphosarcoma lines should, therefore, provide a good model to help elucidate the role of altered myb locus and other protooncogenes in myelomonocytic leukemia. Contrary to studies on induced differentiation of myelomonocytic cells with normal myb locus, our results on plasmacytoid lymphosarcoma cells indicate that reduced myb and myc expression may not be obligatory for growth arrest to occur. The present study, however, supports the previous notion that the myb transforming ability may be restricted to cells of the myelomonocytic lineage. In addition, we found that only the more mature cells can undergo prolonged phorbol 12-myristate 13-acetate-induced growth arrest, suggesting that the maintenance of these leukemic cells in their proliferative state, presumably by the myb gene product, can be overcome with appropriate differentiation signal(s). PMID- 3025855 TI - Molecular cloning of Ly-1, a membrane glycoprotein of mouse T lymphocytes and a subset of B cells: molecular homology to its human counterpart Leu-1/T1 (CD5). AB - We report the isolation of cDNA clones of the mouse lymphocyte differentiation antigen Ly-1. One of these cDNA clones was confirmed to be full-length by DNA sequencing and by expression of Ly-1 by L cells transfected with this clone. Analysis of the predicted amino acid sequence indicated that the Ly-1 polypeptide is synthesized with a 23 amino acid leader and that the mature protein consists of an amino-terminal region of 347 amino acids, a transmembrane sequence of 30 residues, and a carboxyl-terminal region of 94 amino acids. The amino-terminal region appears to be divided into two subregions by a threonine- and proline-rich sequence of 23 amino acids that is highly conserved between Ly-1 and its human homologue Leu-1 (CD5) in position and amino acid composition. The first amino terminal subregion of 111 amino acids is predicted to be arranged in a beta pleated sheet structure of six strands. The entire amino-terminal region is rich in cysteine, with all of its 22 cysteine residues conserved between Ly-1 and Leu 1. The carboxyl-terminal region has no cysteines. Ly-1 and Leu-1 are 63% identical, with a gradient of identical residues from 43% for the first amino terminal subregion to 58% for the second amino-terminal subregion and 90% for the carboxyl-terminal region. The predicted secondary structure of the first amino terminal subregion and identities of certain conserved residues among most members of the immunoglobulin gene superfamily suggest that Ly-1 and Leu-1 are distant members of this family. PMID- 3025857 TI - Multiple nuclear proteins bind upstream sequences in the promotor region of a T cell receptor beta-chain variable-region gene: evidence for tissue specificity. AB - DNA-nuclear protein binding interactions were examined in the promoter region of a representative T-cell receptor Ti beta-chain variable-region gene by means of electrophoretic mobility-shift and DNase I-protection analysis. Within 175 bases upstream of the transcription initiation site, four protected regions ("footprints") were identified on the coding strand, at nucleotides -46 to -68 (I), -72 to -92 (II), -113 to -134 (III), and -136 to -175 (IV). Nuclear proteins (0.6 M NaCl fraction from a heparin-Sepharose column chromatography of nuclear extracts) of a variety of cell types produced footprints I, III, and IV and a fifth footprint (beyond nucleotide -200). In contrast, footprint II was produced only by T-cell extracts, although nuclear extracts of a transformed B lymphoblastoid line produced a partial footprint in this region. Furthermore, footprint analysis of the noncoding strand showed that a continuous region of protection corresponding to the entire region of footprints I and II was generated, along with a DNase I-hypersensitive site, by nuclear proteins derived from T cells but not other cell types. Footprint I has the sequence structure of many well-defined protein-DNA binding sites. Nucleotide sequences in the region of footprint II bore no homology to known sequences, whereas those in the areas of footprints III and IV were similar to motifs within immunoglobulin and other enhancers. These findings may have implications for the tissue specificity of human Ti beta-chain gene transcription. PMID- 3025856 TI - Chimeric antibody with human constant regions and mouse variable regions directed against carcinoma-associated antigen 17-1A. AB - We have cloned the genomic DNA fragments encoding the heavy and light chain variable regions of monoclonal antibody 17-1A, and we have inserted them into mammalian expression vectors containing genomic DNA segments encoding human gamma 3 and kappa constant regions. The transfer of these expression vectors containing mouse-human chimeric immunoglobulin genes into Sp2/0 mouse myeloma cells resulted in the production of functional IgG that retained the specific binding to the surface antigen 17-1A expressed on colorectal carcinoma cells. PMID- 3025858 TI - Suppressive effect on polyclonal B-cell activation of a synthetic peptide homologous to a transmembrane component of oncogenic retroviruses. AB - Purified feline leukemia virus, UV light-inactivated feline leukemia virus, and a synthetic peptide (CKS-17) homologous to a well-conserved region of the transmembrane components of several human and animal retroviruses were each studied for their effects on IgG production by feline peripheral blood lymphocytes. Using a reverse hemolytic plaque assay, both the viable virus and the UV-inactivated feline leukemia virus, but not the CKS-17, activated B lymphocytes to secrete IgG. When staphylococcal protein A, a polyclonal B-cell activator, was used to stimulate IgG synthesis by feline lymphocytes, the viable virus, the UV-inactivated virus, and the CKS-17 peptide each strongly suppressed IgG secretion without compromising viability of the lymphocytes. These findings suggest that the immunosuppressive influences of feline leukemia virus on immunoglobulin synthesis may reside in a conserved portion of the envelope glycoprotein that includes the region homologous to CKS-17. PMID- 3025859 TI - Molecular analysis of TCRB and ABL in a t(7;9)-containing cell line (SUP-T3) from a human T-cell leukemia. AB - A translocation between chromosomes 7 and 9, t(7;9), has been described in cell lines derived from the malignant cells of children with acute T-cell lymphoblastic leukemia or lymphoma. Our cytogenetic analysis of one such cell line, SUP-T3, demonstrates that the breakpoints on chromosomes 7 and 9 lie within bands q36 and q34, respectively, corresponding to the location of the gene encoding the beta chain of the T-cell receptor, TCRB, and the gene homologous to the transforming gene of the Abelson murine leukemia virus, ABL. We investigated the role of these genes in the t(7;9). In situ chromosomal hybridization of TCRB and ABL probes to metaphase cells from SUP-T3 demonstrated that ABL is translocated from chromosome 9 to 7 and that all or part of TCRB is translocated from chromosome 7 to 9. Southern blot analysis revealed that both TCRB alleles were rearranged; however, it could not be determined whether the translocation breakpoint lies within this gene. Pulsed-field gel electrophoresis and Southern blot analysis were used to examine more than 500 kilobases of the ABL locus; we concluded that there are no rearrangements within 250 kb in either direction of the sequences homologous to v-abl. Additionally, no abnormal ABL protein was detected in an in vitro phosphorylation assay. These results indicate that, in SUP-T3, the breakpoint on chromosome 9 lies proximal to ABL and that the break results in no apparent alteration of the ABL protein. We therefore hypothesize that another gene on chromosome 9, at band q34, plays a role in this translocation. This study also demonstrates that pulsed-field gel electrophoresis is a powerful new tool for the analysis of human chromosomal translocations. PMID- 3025860 TI - Immunoprecipitation of the parathyroid hormone receptor. AB - An 125I-labeled synthetic analog of bovine parathyroid hormone, [8-norleucine,18 norleucine,34-tyrosine]PTH-(1-34) amide ([Nle]PTH-(1-34)-NH2), purified by high pressure liquid chromatography (HPLC), was employed to label the parathyroid hormone (PTH) receptor in cell lines derived from PTH target tissues: the ROS 17/2.8 rat osteosarcoma of bone and the CV1 and COS monkey kidney lines. After incubation of the radioligand with intact cultured cells, the hormone was covalently attached to receptors by using either a photoaffinity technique or chemical (affinity) cross-linking. In each case, covalent labeling was specific, as evidenced by a reduction of labeling when excess competing nonradioactive ligand was present. After covalent attachment of radioligand, membranes were prepared from the cells and solubilized in the nonionic detergent Nonidet P-40 or octyl glucoside. The soluble membrane fraction present in the supernatant of a 100,000 X g centrifugation was incubated with IgG prepared from anti-PTH antiserum generated to the amino-terminal region, residues 1-34, of PTH. The IgG PTH-receptor complex was precipitated with staphylococcal protein A-Sepharose. Analysis of the immunoprecipitate on NaDod-SO4/polyacrylamide gel electrophoresis followed by autoradiography revealed the presence of a doublet of apparent molecular mass 69-70 kDa. Specifically labeled bands of approximate molecular mass 95 and 28 kDa were also observed. The anti-PTH IgG was affinity purified by passage over a PTH-Sepharose column and used to make an immunoaffinity column. The 70- and 28-kDa bands were also observed after labeled solubilized membrane preparations were allowed to bind to this column and then were eluted by using a [Nle]PTH-(1-34)-NH2-containing buffer or acetic acid. These studies suggest that the use of an anti-PTH antiserum that binds receptor-bound hormone is likely to be a useful step in the further physiochemical characterization and purification of the PTH receptor. PMID- 3025861 TI - Involvement of a specific protein in the regulation of a circadian rhythm in Aplysia eye. AB - Our previous results indicated that protein synthesis was necessary for serotonin (5-hydroxytryptamine, 5-HT) to regulate the phase of the biological clock in the Aplysia eye. Also, we showed that 5-HT appeared to increase the synthesis of a 34 kDa protein with an isoelectric point of 7.2. Subsequent studies were carried out to quantitate the effect of 5-HT on the 34-kDa protein and to examine whether the 34-kDa protein was involved in the circadian timing system. The regional specificity of the effect of 5-HT on the 34-kDa protein was investigated. The proximal portion of the eye appeared to synthesize much more of the 34-kDa protein than the distal portion. Also, 5-HT had a much larger effect on the synthesis of the 34-kDa protein in the proximal portion than in the distal portion. The proximal location of synthesis and the 5-HT effect on the synthesis of the 34-kDa protein correlate with the proximal location of cells and processes that are necessary for the expression of the circadian rhythm. The relationship between the effect of 5-HT on the circadian rhythm and the effect of 5-HT on the 34-kDa protein was also examined. As 5-HT causes phase shifts in the rhythm by activating adenylate cyclase to increase cAMP, forskolin and 8 benzylthioadenosine 3',5'-cyclic monophosphate mimicked the effect of 5-HT on the 34-kDa protein. We also found that 5-HT significantly increased the synthesis of the 34-kDa protein at phases when 5-HT delays or advances the phase of the rhythm but did not increase the synthesis of the 34-kDa protein at a phase when 5-HT did not phase shift. This phase-dependent effect of 5-HT on the 34-kDa protein qualitatively accounts for the phase dependence of the effect of 5-HT on the circadian rhythm. These results, when considered together with our earlier data, suggest that the synthesis of the 34-kDa protein is directly involved in the phase shift produced by 5-HT. The 34-kDa protein is worthy of future investigation as a candidate for a component of the circadian oscillator. PMID- 3025862 TI - Organization of the neural cell adhesion molecule (N-CAM) gene: alternative exon usage as the basis for different membrane-associated domains. AB - The neural cell adhesion molecule, N-CAM, is expressed as at least three polypeptide chain, (ld, sd, and ssd chains) specified by a single gene and derived by alternative splicing and polyadenylation-site selection during RNA processing. We describe here the characterization of seven overlapping genomic phage clones reactive with N-CAM cDNA, indicating that the chicken N-CAM gene is more than 50 kilobases long. Analysis of the gene shows that there are at least 19 exons and that the coding sequences for the ld, sd, and ssd chains are assembled from 18, 17, and 15 exons, respectively. The first 14 exons appear to be common to all three chains and encode the amino-terminal portion of N-CAM, which contains five tandem homologous repeats resembling those seen in the immunoglobulin gene superfamily. In contrast to other genes containing such domains, each of these segments in N-CAM is specified by two exons. The carboxyl terminal portion of each N-CAM chain is different as a result of the alternative use of exons. A single exon encodes the carboxyl-terminal 26 amino acids of the ssd chain and the 3' untranslated region of its mRNA, ending with a poly(A) addition site. Two exons encode the transmembrane and cytoplasmic sequences common to the ld and sd chains, and another exon encodes the additional 261 amino acids found in the cytoplasmic domain of the ld chain. The carboxyl-terminal 21 amino acids common to the ld and sd chains and the 3' untranslated region common to their mRNAs are encoded by a single large exon of 3475 base pairs that ends with a second poly(A)-addition site. Sequences from the 13-kilobase intron that separates the exons encoding the amino-terminal and carboxyl-terminal regions of the molecule hybridize to a 2-kilobase poly(A)+ RNA transcript of unknown identity. This description of the chicken N-CAM gene provides a basis for determining the mechanisms that regulate the differential expression of the N-CAM polypeptide chains during development. PMID- 3025863 TI - cDNA for the human beta 2-adrenergic receptor: a protein with multiple membrane spanning domains and encoded by a gene whose chromosomal location is shared with that of the receptor for platelet-derived growth factor. AB - We have isolated and sequenced a cDNA encoding the human beta 2-adrenergic receptor. The deduced amino acid sequence (413 residues) is that of a protein containing seven clusters of hydrophobic amino acids suggestive of membrane spanning domains. While the protein is 87% identical overall with the previously cloned hamster beta 2-adrenergic receptor, the most highly conserved regions are the putative transmembrane helices (95% identical) and cytoplasmic loops (93% identical), suggesting that these regions of the molecule harbor important functional domains. Several of the transmembrane helices also share lesser degrees of identity with comparable regions of select members of the opsin family of visual pigments. We have localized the gene for the beta 2-adrenergic receptor to q31-q32 on chromosome 5. This is the same position recently determined for the gene encoding the receptor for platelet-derived growth factor and is adjacent to that for the FMS protooncogene, which encodes the receptor for the macrophage colony-stimulating factor. PMID- 3025865 TI - Solubilization of a membrane-bound diol dehydratase with retention of EPR g = 2.02 signal by using 2-(N-cyclohexylamino)ethanesulfonic acid buffer. AB - A procedure for solubilization of the oxygen-labile, membrane-bound diol dehydratase from Clostridium glycolicum with retention of enzymatic activity is described. The procedure involves sonication of crude membrane preparations anaerobically in 0.1 M 2-(N-cyclohexylamino)ethane-sulfonic acid (CHES) buffer (pH 8.6-9.0) containing 2 mM dithiothreitol. The addition of dimethylsulfoxide (30%) and lysophosphatidylcholine (0.15 mg/ml) to the solubilization buffer resulted in a 10-fold increase in recovery of solubilized diol dehydratase activity. After ultracentrifugation, an overall recovery of 50% of the activity initially present in the crude membrane preparations was achieved. Active membrane preparations and the solubilized enzyme exhibited an EPR signal at g = 2.02. Both enzyme activity and EPR signal were sensitive to oxygen and the radical scavengers, NH2OH and hydroxy-urea. PMID- 3025864 TI - Host cell proteins bind to the cis-acting site required for virion-mediated induction of herpes simplex virus 1 alpha genes. AB - The herpes simplex virus 1 genes form at least five groups (alpha, beta 1, beta 2, gamma 1, and gamma 2) whose expression is coordinately regulated and sequentially ordered in a cascade fashion. In productively infected cells, the alpha genes are expressed first, and a virion protein, the alpha-trans-inducing factor (alpha-TIF), acts in trans to enhance their expression. Induction of the alpha genes by alpha-TIF requires the presence of a trans-induction cis-acting site (alpha-TIC), and one to three homologs of the alpha-TIC sequence are contained in the regulatory domains of all alpha genes. We report that small DNA fragments from regulatory domains of alpha 0, alpha 4, and alpha 27 genes containing alpha-TIC homologs formed complexes with host but not viral proteins. DNase protection studies indicated that the major host protein complex alpha-H1 detected in DNA gel retardation assays bound asymmetrically across the alpha-TIC site. All DNA fragments containing alpha-TIC homologs, but not those lacking the homolog, competed for the binding of this complex. The location of the binding site of the other host proteins is not yet known. Simian virus 40 DNA fragments containing a homolog of the alpha-TIC sequence also competed with herpes simplex virus DNA fragments carrying authentic alpha-TIC homologs for the alpha-H1 protein complex. PMID- 3025866 TI - NH2-terminal sequences of two src proteins that cause aberrant transformation. AB - Two isolates of recovered avian sarcoma viruses (rASVs), rASV157 and rASV1702, transform cells in culture, but have greatly reduced in vivo tumorigenicity. The src proteins of rASV157 and rASV1702 have alterations within their NH2 termini, are not myristoylated, and have an altered subcellular localization. We have molecularly cloned and determined the nucleotide sequences of the src genes of rASV157 and rASV1702. We found that their src proteins have unusual NH2 termini: the rASV157 src protein NH2 terminus consists of 30 amino acids of the env signal peptide attached to Ser-6 of the src sequence, while the rASV1702 src protein NH2 terminus consists of 45 amino acids of the env signal peptide attached to Ala-76 of the src sequence. Expression of recombinant Rous sarcoma virus constructs containing the molecularly cloned rASV src genes produced src proteins with the same properties as those of the parental viruses. Our results suggest that the NH2-terminal structures are responsible for many unusual properties of the mutant src proteins. PMID- 3025867 TI - Polylysine-containing peptides, including the carboxyl-terminal segment of the human c-Ki-ras 2 protein, affect the activity of some key membrane enzymes. AB - Polylysine-containing peptides are found to affect membrane protein kinases, phosphatidylinositol kinases, and adenylate cyclase. Poly(L-lysine), poly(D lysine), random copolymers of lysine and serine or lysine and alanine, and poly(L ornithine) produced large increases in the in vitro phosphorylation of some membrane proteins present in Xenopus laevis oocyte membranes. Poly(L-arginine) did not cause a similar stimulation. In these membranes the phosphorylation of polydisperse protein of approximately 25 kDa was also greatly increased by 1 mM spermine and spermidine, by 10 microM histone H1, or by 200 microM peptide containing the 14-residue sequence at the carboxyl terminus of the human c-Ki-ras 2 gene product, which has eight lysines. Similar specific stimulation of protein phosphorylation was observed with membranes of NG-108-15 nerve cells in culture. Polylysine peptides, including the c-Ki-ras 2 segment, also stimulate the in vitro phosphorylation of membrane inositolphospholipids, to produce mainly phosphatidylinositol 4-phosphate and less phosphatidylinositol 4,5-bisphosphate. Polylysine also alters the activity of oocyte adenylate cyclase, assayed in the presence of either F- or 5'-guanylyl imidodiphosphate. PMID- 3025868 TI - Delta-crystallin genes become hypomethylated in postmitotic lens cells during chicken development. AB - Although it has been argued that the loss of 5-methylcytosine from specific sites in DNA plays an important role in activation of specific genes, the mechanism of hypomethylation is not well understood. One model links the process to DNA replication, proposing that it occurs by not remethylating cytosine on newly synthesized DNA. An alternative model argues that hypomethylation results from excision of part or all of the 5-methylcytosine. We were able to test whether hypomethylation can occur without replication by analysis of methylation changes in the delta-crystallin genes of the chicken lens. During embryonic development a large fraction of cells in the lens stops dividing as part of the differentiation process. Shortly after this stage, the delta-crystallin genes in samples of the whole lens become hypomethylated, suggesting the possibility that this process might be occurring in the subset of cells that is no longer dividing. We found that hypomethylation of these genes does occur in postmitotic lens cells, a result that implicates an excision mechanism in this tissue. PMID- 3025869 TI - Phosphorylation by protein kinase C of a 20-kDa cytoskeletal polypeptide enhances its susceptibility to digestion by calpain. AB - Incubation of the cytoskeletal fraction from human neutrophils with the proteolytically activated form of protein kinase C results in the phosphorylation of several components, including a 20-kDa polypeptide, probably consisting of myosin light chains. The 20-kDa polypeptide is also specifically phosphorylated by activated protein kinase C in a solubilized 20-kDa/80-kDa complex that was obtained after sonication of the insoluble cytoskeletal fraction. Phosphorylation of this polypeptide, in either the insoluble cytoskeletal fraction or the soluble 20-kDa/80-kDa complex, greatly enhances its susceptibility to digestion by the Ca2+-requiring proteinase (calpain, EC 3.4.22.17) of human neutrophils. Thus, signals that activate calpain by mobilizing intracellular calcium would lead to proteolytic activation of protein kinase C, phosphorylation of cytoskeletal proteins, and remodeling of the cytoskeleton by proteolysis of at least one cytoskeletal component. PMID- 3025870 TI - Cytochrome P450c17 (steroid 17 alpha-hydroxylase/17,20 lyase): cloning of human adrenal and testis cDNAs indicates the same gene is expressed in both tissues. AB - P450c17 is the single enzyme mediating both 17 alpha-hydroxylase (steroid 17 alpha-monooxygenase, EC 1.14.99.9) and 17,20 lyase activities in the synthesis of steroid hormones. It has been suggested that different P450c17 isozymes mediate these activities in the adrenal gland and testis. We sequenced 423 of the 509 amino acids (83%) of the porcine adrenal enzyme; based on this partial sequence, a 128-fold degenerate 17-mer was synthesized and used to screen a porcine adrenal cDNA library. This yielded a 380-base cloned cDNA, which in turn was used to isolate several human adrenal cDNAs. The longest of these, lambda hac17-2, is 1754 base pairs long and includes the full-length coding region, the complete 3' untranslated region, and 41 bases of the 5'-untranslated region. This cDNA encodes a protein of 508 amino acids having a predicted molecular weight of 57,379.82. High-stringency screening of a human testicular cDNA library yielded a partial clone containing 1303 identical bases. RNA gel blots and nuclease S1 protection experiments confirm that the adrenal and testicular P450c17 mRNAs are indistinguishable. These data indicate that the testis possesses a P450c17 identical to that in the adrenal. The human amino acid sequence is 66.7% homologous to the corresponding regions of the porcine sequence, and the human cDNA and amino acid sequences are 80.1 and 70.3% homologous, respectively, to bovine adrenal P450c17 cDNA. Both comparisons indicate that a central region comprising amino acid residues 160-268 is hypervariable among these species of P450c17. Comparison of the amino acid sequence of P450c17 with two other human steroidogenic cytochromes P450 show much greater homology with P450c21 (28.9%), another microsomal enzyme, than with P450scc (12.3%), a mitochondrial enzyme. PMID- 3025871 TI - Dependence of thermodynamic efficiency of proton pumps on frequency of oscillatory concentration of ATP. AB - In order to evaluate the utilization of variable ATP concentration produced by an oscillatory reaction (as in anaerobic glycolysis), we analyze the thermodynamic efficiency of power output of a cyclic, ATP-driven proton pump found in the plasma membrane of plant cells. The model used includes the coupling of potassium and calcium ion transport. Oscillations in the concentration of ATP can lead to either increases or decreases in efficiency compared to that at constant ATP concentration, with corresponding decreases and increases in dissipation in the irreversible processes of the proton pump, depending on the frequency of the oscillations. Variations of imposed frequencies induce, in the periodic response, variations of phase shifts between the components of the total membrane current, which consist of the pump's proton current and the currents of potassium and calcium ions. Increases in efficiency are attained when the phase shifts are such that maxima (or minima) in the proton pump current and membrane potential occur simultaneously. PMID- 3025872 TI - Stable expression and replication of hepatitis B virus genome in an integrated state in a human hepatoma cell line transfected with the cloned viral DNA. AB - A human hepatocellular carcinoma cell line (Huh6-c15) was transfected with a recombinant DNA molecule that consists of tandemly arranged hepatitis B virus (HBV) genome and a neomycin-resistant gene. One clone resistant to G-418 produces and releases surface antigen and e antigen into medium at a high level and accumulates core particles intracellularly. This clone has a chromosomally integrated set of the original recombinant DNA and produces a 3.5-kilobase transcript corresponding to the pregenome RNA as well as HBV DNAs in an extrachromosomal form. Most of these DNAs were in single-stranded or partially double-stranded form and were packaged in the intracellular core particles. In the medium, particles were detected that contained HBV DNA and were morphologically indistinguishable from Dane particles. These results demonstrate that the HBV genome in an integrated state acted as a template for viral gene expression and replication. The cells were maintained for more than 6 months without losing the ability to produce the extrachromosomal HBV DNA and Dane-like particles. Thus, the cells can be used as a model system for analyses of gene expression and DNA replication of HBV in human hepatocytes. PMID- 3025873 TI - A pancreas specificity results from the combination of polyomavirus and Moloney murine leukemia virus enhancer. AB - An infectious recombinant polyomavirus was constructed in which a regulatory region of its genome, the B enhancer region (nucleotides 5128-5265) has been replaced with the 72- or 73-base-pair repeat enhancer from the Moloney murine leukemia virus genome. We show that this recombinant polyomavirus displays a strong tissue specificity for the pancreas of mice. This organ was not permissive for either the parental polyomavirus, which is predominantly kidney and salivary gland specific, or the Moloney murine leukemia virus, which is lymphotropic. This result indicated that tissue specificity can be achieved by a combination of apparently modular elements. Some of the implications of a modular mechanism of tissue specificity are considered. PMID- 3025874 TI - Cytoplasts made from human blood polymorphonuclear leukocytes with or without heat: preservation of both motile function and respiratory burst oxidase activity. AB - Anucleate fragments (cytoplasts) from polymorphonuclear leukocytes (PMN) are simplified systems that can be used to elucidate specific pathways by which cell function is altered. PMN cytoplasts in current use are defective either in activatable respiratory burst oxidase activity or in motile function. By centrifugation of PMN on discontinuous gradients of Ficoll without cytochalasin B, we have created granule-poor cytoplasts in which both these capacities are preserved. Specifically, they generate superoxide anion (O2-.) and reduce nitroblue tetrazolium dye on appropriate stimulation; they respond chemotactically to erythrocytes destroyed by laser microirradiation or to the specific chemoattractants fMet-Leu-Phe (10 nM) and C5a (zymosan-activated serum); and they ingest and kill staphylococci. We can improve the yield of these fragments progressively by preheating (45 degrees C) the cells in suspension for increasing periods of time, but those treatments are attended by a decreasing percentage of cytoplasts with activatable oxidase activity, and a progressive inability of the cytoplasts to ingest and to kill staphylococci. These easily made and multipotent cytoplasts readily lend themselves to studies of PMN physiology. PMID- 3025875 TI - Structural features of cartilage matrix protein deduced from cDNA. AB - cDNAs encoding the Mr 54,000 chicken cartilage matrix protein (CMP) were selected from a cartilage cDNA expression library by immunological means. Antibodies elicited against insert-encoded protein purified from one of the clones reacted specifically with chicken CMP in immunoblots of total cartilage extract, providing positive identification of the cDNA clones isolated. The cDNAs detect a 3.4-kilobase transcript that was present in sternal cartilage and in cartilaginous but not in precartilaginous embryonic limb tissues. The cDNAs code for 416 amino acids of the chicken CMP, including its COOH terminus. There are two striking features in the deduced CMP amino acid sequence: first, it contains a region with significant homologies to repeat sequences in the precursor for epidermal growth factor; and second, it is made up of two large homologous repeat sequences. These results provide the first detailed structural information on the CMP and establish it as a developmentally regulated marker of cartilage differentiation. PMID- 3025876 TI - Evolution of the genus Leishmania as revealed by comparisons of nuclear DNA restriction fragment patterns. AB - Restriction endonuclease DNA fragment patterns have been used to examine the relationships among 28 isolates of Leishmania as well as Crithidia, Endotrypanum, and Trypanosoma cruzi. Fragments of nuclear DNA were generated with six restriction enzymes, and blots were hybridized with probes from three loci. Among the major lineages the fragment patterns are essentially completely different, while within the major lineages various degrees of divergence are found. Molecular evolutionary trees were constructed using the method of Nei and Li to estimate the percent nucleotide sequence divergence among strains from the fraction of fragments shared. Defined groups, such as species or subspecies within the major lineages, are also grouped by nuclear DNA comparisons. Within the donovani complex, we find Leishmania donovani chagasi and Leishmania donovani infantum to be as similar as strains within Leishmania donovani donovani, consistent with the proposal by other workers that New World visceral leishmaniasis originated quite recently. PMID- 3025877 TI - A locus for a human hereditary cataract is closely linked to the gamma-crystallin gene family. AB - Within the human gamma-crystallin gene cluster polymorphic Taq I sites are present. These give rise to three sets of allelic fragments from the gamma crystallin genes. Together these restriction fragment length polymorphisms define eight possible haplotypes, three of which (Q, R, and S) were found in the Dutch and English population. A fourth haplotype (P) was detected within a family in which a hereditary Coppock-like cataract of the embryonic lens nucleus occurs in heterozygotes. Haplotype P was found only in family members who suffered from cataract, and all family members who suffered from cataract had haplotype P. The absolute correlation between the presence of haplotype P and cataract within this family shows that the gamma-crystallin gene cluster and the locus for the Coppock like cataract are closely linked [logarithm of odds (lod) score of 7.58 at its maximum at phi = 0]. This linkage provides genetic evidence that the primary cause of a cataract in humans could possibly be a lesion in a crystallin gene. PMID- 3025879 TI - his operons of Escherichia coli and Salmonella typhimurium are regulated by DNA supercoiling. AB - The hisW mutations of Salmonella typhimurium are highly pleiotropic mutations that elevate his operon expression, reduce ilv gene expression, alter stable RNA metabolism, and confer defective growth properties. The hisW mutations are highly linked to a naladixic acid-resistant gyrA mutation of S. typhimurium. Multicopy recombinant plasmids containing the Escherichia coli gyrA gene are able to complement both the growth defects and the elevated his operon expression associated with the hisW mutations. We conclude that hisW mutations are alleles of the gyrA gene. The hisU1820 mutant of S. typhimurium exhibits many of the same phenotypes as hisW mutants. Several lines of evidence, including high transduction linkage to recF, suggest that hisU1820 is an allele of gyrB. Finally, well-characterized gyrA and gyrB alleles of E. coli are also his regulatory mutations. We propose that a wild-type degree of chromosomal superhelicity is required for maximal production of histidyl-tRNA and normal his operon regulation. PMID- 3025878 TI - Human collagen genes encoding basement membrane alpha 1 (IV) and alpha 2 (IV) chains map to the distal long arm of chromosome 13. AB - At least 20 genes encode the structurally related collagen chains that comprise greater than 10 homo- or heterotrimeric types. Six members of this multigene family have been assigned to five chromosomes in the human genome. The two type I genes, alpha 1 and alpha 2, are located on chromosomes 17 and 7, respectively, and the alpha 1 (II) gene is located on chromosome 12. Our recent mapping of the alpha 1 (III) and alpha 2 (V) genes to the q24.3----q31 region of chromosome 2 provided the only evidence that the collagen genes are not entirely dispersed. To further determine their organization, we and others localized the alpha 1 (IV) gene to chromosome 13 and in our experiments sublocalized the gene to band q34 by in situ hybridization. Here we show the presence of the alpha 2 type IV locus also on the distal long arm of chromosome 13 by hybridizing a human alpha 2 (IV) cDNA clone to rodent-human hybrids and to metaphase chromosomes. To our knowledge, these studies represent the only demonstration of linkage between genes encoding both polypeptide chains of the same collagen type. PMID- 3025880 TI - Kinetoplast DNA minicircles: regions of extensive sequence divergence. AB - Previous work has shown that the kinetoplast minicircle DNA of Leishmania species exhibits species-specific sequence divergence and this observation has led to the development of a DNA probe-based diagnostic test for leishmaniasis. In the work reported here, we demonstrate that the minicircle is composed of three types of DNA sequences with differing specificities reflecting different rates of DNA sequence change. A library of cloned fragments of kinetoplast DNA (kDNA) from Leishmania mexicana amazonensis was prepared and the cloned subfragments were found to contain DNA sequences with different taxonomic specificities based on hybridization analysis with various species of Leishmania. Four groups of subfragments were found, those that hybridized with a large number of Leishmania sp. as well as sequences unique to the species, subspecies, or isolate. Analysis of nested deletions of a single, full-length minicircle demonstrates that these different taxonomic specificities are contained within a single minicircle. This implies that different regions of a single minicircle have DNA sequences that diverge at different rates. These sequences represent potentially valuable tools in diagnostic, epidemiologic, and ecological studies of leishmaniasis and provide the basis for a model of kDNA sequence evolution. PMID- 3025881 TI - Antibody responses to Epstein-Barr virus-determined nuclear antigen (EBNA)-1 and EBNA-2 in acute and chronic Epstein-Barr virus infection. AB - Five distinct Epstein-Barr virus (EBV)-determined nuclear antigens (EBNA-1 to EBNA-5) were recently identified. Antibody responses to these antigens could conceivably differ, and thus prove of serodiagnostic value, in EBV-associated disease processes. As a first step, murine or human cell lines transfected with appropriate EBV DNA fragments and stably expressing either EBNA-1 or EBNA-2 were used to determine the frequency and time of emergence of antibodies to these two antigens in the course of acute and chronic infectious mononucleosis (IM) and to assess their titers in so-called chronic active EBV infections. Following IM, antibodies to EBNA-2 arose first and, after reaching peak titers, declined again in time to lower persistent or even nondetectable levels. Antibodies to EBNA-1 emerged several weeks or months after anti-EBNA-2 and gradually attained the titers at which they persisted indefinitely. The ratios between the anti-EBNA-1 and anti-EBNA-2 titers therefore were generally well below 1.0 during the first 6 12 months after IM and turned to well above 1.0 during the second year. In clear cases of chronic IM, the inversion of this ratio was delayed or prevented. In the less well-defined chronic EBV infections, low ratios were observed in only some of the patients. Because many of these illnesses were not ushered in by a proven IM and often showed EBV-specific antibody profiles within the normally expected range, a causal role of the virus in these cases remains doubtful. PMID- 3025883 TI - Regulation of phagocyte function by alpha-tocopherol. PMID- 3025882 TI - Structure and expression of the gene encoding the vasoactive intestinal peptide precursor. AB - The gene encoding the human vasoactive intestinal peptide (VIP) and the histidine methionine amide (PHM-27) peptide hormone was isolated from lambda phage libraries. The human gene was found to be composed of seven exons spanning approximately 9 kilobase pairs. The first exon codes for an untranslated leader sequence, and the second exon codes for a putative signal peptide. DNA sequences coding for the VIP and PHM-27 hormones are located in two different exons. Southern blot analysis with genomic DNA suggested that a single copy of the VIP/PHM-27 gene is present in the human haploid genome. The expression of VIP/PHM 27 precursor mRNA in various tissues in the rat was analyzed by RNA gel blot hybridization. In the organs examined, expression was only detected in the brain and duodenum. RNA isolated from various regions of the rat brain--including the cortex, hypothalamus, and hippocampus--hybridized to both VIP- and PHM-27 specific probes. The same pattern of hybridization was found when VIP- and PHM-27 specific probes were used, suggesting that possible differences in the localization of VIP and PHM-27 peptides between different brain regions cannot be accounted for by differential RNA processing. PMID- 3025884 TI - Epidemiologic studies of diet and cancer. PMID- 3025885 TI - Calorie restriction, ad libitum feeding, and cancer. AB - The inhibition of cancer by calorie restriction was discovered over 50 years ago. By 1950 it had been well characterized and there existed sufficient data to propose a mechanism of action. For reasons that remain unclear, but are probably related to the perception of the calorie restricted rodent as "small" and the ad libitum feeding regimen as more "normal," the concept of calorie restriction has been largely ignored by investigators after this time. Hence, despite the fact that calorie restriction is one of the oldest, best-documented, and most effective ways known to reduce cancer risk in rodents, it has had little impact on modern cancer research. In this report the history of calorie restriction is briefly reviewed, and a mechanism of action is proposed that involves increased production of ACTH and decreased production of gonadotrophins. It is further proposed that these changes may come about in part from the restriction of the time during which feeding is permitted as well as from the restriction of food per se. There is renewed interest in calorie restriction due in part to the growing recognition that there are differences in the efficiency of utilization of various sources of energy, in particular that fat calories are utilized more efficiently and provide more usable energy than carbohydrate calories. New data are presented indicating that the apparent enhancement by dietary fat of mammary cancer in rats is really a manifestation of the caloric effect. Further, the effect is abolished by moderate calorie restriction of only 15-20%. The application of these findings to the prevention of cancer in humans is considered. PMID- 3025887 TI - Binding of cobalamin analogs to intrinsic factor-cobalamin receptor and its prevention by R binder. AB - It is now known that nonphysiological cobalamin analogs exist in the gastrointestinal tract, but their metabolic behavior is unclear. In this study, [57Co]cobinamide was used to study its affinity to hog intrinsic factor-cobalamin (IF-Cbl) receptor which has no species specificity against human IF-Cbl receptor, and its relation to human saliva R binder. Cobinamide was prepared from [57Co]cyanocobalamin and separated by paper chromatography. Human IF-Cbl complex was bound to IF-Cbl receptor but free cyanocobalamin was not. Although R binder cobinamide was not bound to the IF-Cbl receptor, free cobinamide was bound to the IF-Cbl receptor to a significant extent (about one-half of IF-cyanocobalamin binding to the IF-Cbl receptor). We then investigated the binding of cobinamide to R binder and trypsin-treated R binder. Association constant of cobinamide binding to the IF-Cbl receptor was 1.0 X 10(9) M-1 which was much lower than that of cobinamide binding to trypsin-treated R binder and to untreated R binder. Further study indicated that cobinamide binding to the IF-Cbl receptor was blocked by the addition of R binder and also by trypsin-treated R binder. We conclude that one of the roles of R binder is to prevent binding of free cobalamin analogs to the IF-Cbl receptor in the gut. PMID- 3025886 TI - Relationship between dietary fiber and cancer: metabolic, physiologic, and cellular mechanisms. AB - The relationships between fiber consumption and human cancer rates have been examined, together with an analysis of the effects of individual dietary fibers on the experimental induction of large bowel cancer. The human epidemiology indicates an inverse correlation between high fiber consumption and lower colon cancer rates. Cereal fiber sources show the most consistent negative correlation. However, human case-control studies in general fail to confirm any protective effect due to dietary fiber. Case-control studies indicate that if any source of dietary fiber is possibly antineoplastic then it is probably vegetables. These results may mean that purified fibers alone do not inhibit tumor development, whereas it is likely that some other factors present in vegetables are antineoplastic. Experiments in laboratory animals, using chemical induction of large bowel cancer, have in general shown a protective effect with supplements of poorly fermentable fibers such as wheat bran or cellulose. In contrast, a number of fermentable fiber supplements including pectin, corn bran, oat bran, undegraded carageenan, agar, psyllium, guar gum, and alfalfa have been shown to enhance tumor development. Possible mechanisms by which fibers may inhibit colon tumorigenesis include dilution and adsorption of any carcinogens and/or promoters contained within the intestinal lumen, the modulation of colonic microbial metabolic activity, and biological modification of intestinal epithelial cells. Dietary fibers not only bind carcinogens, bile acids, and other potential toxins but also essential nutrients, such as minerals, which can inhibit the carcinogenic process. Fermentation of fibers within the large bowel results in the production of short chain fatty acids, which in vivo stimulate cell proliferation, while butyrate appears to be antineoplastic in vitro. Evidence suggests that if dietary fibers stimulate cell proliferation during the stage of initiation, then this may lead to tumor enhancement. Fermentation also lowers luminal pH, which in turn modifies colonic microbial metabolic acidity, and is associated with increased epithelial cell proliferation and colon carcinogenesis. Because dietary fibers differ in their physiochemical properties it has been difficult to identify a single mechanism by which fibers modify colon carcinogenesis. Clearly, more metabolic and physiological studies are needed to fully define the mechanisms by which certain fibers inhibit while others enhance experimental colon carcinogenesis. PMID- 3025888 TI - Gastroenteritis caused by human rotaviruses (serotype three) in a suckling mouse model. AB - The pathogenic potential of human rotaviruses of serotypes 1 through 4 was evaluated in suckling mice. Oral inoculation of three different human rotaviruses of serotype 3 into 5-6 day old CD-1 mice caused disease characterized by diarrhea and dehydration. The mean 50% diarrhea inducing dose (DD50) was 5 X 10(5) pfu. Histopathological examination of small intestines revealed villus epithelial cell vacuolization localized to the distal one-third of the villus. Only Serotype 3 rotaviruses exhibited a rapid phase of viral growth in the intestine between 7 and 12 hours post-inoculation. Larger inocula of rotavirus serotypes 1, 2, and 4 did not cause disease or typical histopathologic changes. However, immunoperoxidase staining for rotavirus antigen was positive in all serotypes tested indicating that infection can occur without apparent disease and is not serotype specific. This convenient in-vivo model can be used to evaluate attenuation of human origin vaccine candidates of serotype 3. PMID- 3025889 TI - Spread of herpes simplex virus in lymph nodes after experimental infection of mice. AB - Herpes simplex virus was frequently isolated from ipsilateral popliteal lymph nodes after percutaneous inoculation of the dorsal face of the footpad, and from ipsi- and contralateral submandibular lymph nodes after percutaneous inoculation of the cheek or the orofacial area of mice. Virus was detected only on very rare occasion in nondraining lymph nodes (inguinal or axillary) or in contralateral popliteal lymph nodes, but was frequently isolated in contralateral lumbar lymph nodes after footpad inoculation. The presence of virus in lymph nodes paralleled or followed the invasion of ipsilateral sensory ganglia and was associated with dissemination of virus in contralateral sensory ganglia after unilateral inoculation. In older mice virus was detected only occasionally in lymph nodes and dissemination of virus in contralateral sensory ganglia was generally not observed. The results suggest that lymphatic spread may contribute to dissemination of virus in contralateral sensory ganglia after unilateral inoculation of mice. PMID- 3025890 TI - Passive protection across subgroups of alphaviruses by hyperimmune non-cross neutralizing anti-Sindbis serum. AB - Extended hyperimmunization of rabbits with Sindbis (SIN) or Semliki Forest (SF) viruses causes the production of antisera that are cross-reactive with virus infected cells in antibody-dependent, complement-mediated cytotoxicity assays but that do not cross-neutralize viruses in vitro. C3H/HeJ mice given gamma globulin fractionated from the extended hyperimmune antiserum against SIN, but not control sera, were protected from challenge by 100 LD50 of SF, a virus which is in a different subgroup than SIN. All mice survived if the gamma globulin was given 24 hr before challenge virus and partial protection occurred if the globulin was given 24 hr after the virus. Cobra venom factor treatment of normal C3H mice challenged with SF did not reduce the protection, suggesting that complement was not involved. Methyl palmitate (40 mg/mouse) given before gamma globulin and virus challenge suppressed macrophage activity and reduced the level of protection 23% in females and 70% in males. Silica treatment (3 mg/mouse) reduced the protection equally in both males and females by 92%. In vitro experiments were done to test if it were possible that cross-antibody-dependent cellular cytotoxicity (ADCC) could account for the passive cross-protection observed in this system. Cross-ADCC could be demonstrated in vitro at high dilutions of antiserum (1:25,600). On the basis of the in vitro and in vivo results presented, we suggest that cross-ADCC against SF-infected target cells is one of the likely mechanisms to explain the passive cross-protection observed. PMID- 3025892 TI - Solvents and peripheral neuropathy. PMID- 3025891 TI - Glycosaminoglycans and a newly purified aortic chondroitin proteoglycan block polycationic modulation of protein phosphatase activity. AB - Recently, we described a bovine aortic phosphatase which we called PCM phosphatase (polycation modulable) because its activity in vitro can be modulated by polycations such as polylysine and histone-H1 (Di Salvo J, Gifford D, Kokkinakis A. Modulation of aortic protein phosphatase activity by polylysine. Proc Soc Exp Biol Med 177:24-32, 1984). We We suspected that polycationic modulation might be inhibited by polyanionic glycosaminoglycans. Accordingly, an aortic anionic substance was purified by sequential steps including (a) heating aortic extracts at 90 degrees C, (b) precipitation of protein with (NH4)2 SO4, and (c) anionic-exchange chromatography on a Mono Q HR 5/5 column using the Pharmacia fast protein liquid chromatography system. Electrophoresis (polyacrylamide-agarose) of the purified substance revealed one band which stained metachromatically with toluidine blue; however, no staining occurred with Coomassie blue. Electrophoretic mobility increased following proteolytic digestion of the substance with papain. The substance produced concentration dependent reversal of polylysine-mediated inhibition of myosin light chain dephosphorylation, and it also reversed polylysine-mediated stimulation of phosphorylase phosphatase activity expressed by PCM-phosphatase. Its ability to inhibit or reverse polycationic modulation was abolished after incubation with either chondroitinase AC or chondroitinase ABC. Based on these properties the substance was identified as a chondroitin proteoglycan. Commercially available glycosaminoglycans (heparin and chondroitin sulfates) also reversed polycationic modulation. The results show that modulation of phosphatase activity may be significantly modified by naturally occurring glycosaminoglycans. These studies may also have an important bearing on the purported roles of phosphatase(s) and glycosaminoglycans in calcification of soft tissues. PMID- 3025893 TI - Evidence for the involvement of sulfidopeptide leukotrienes in the pathogenesis of Pichinde virus infection in strain 13 guinea pigs. AB - Strain 13 guinea pigs inoculated subcutaneously with Pichinde virus developed fever, lost body weight, decreased water and food consumption, and died at 14 +/- 0.6 days. After FPL-55712, a leukotriene D4 antagonist, was administered beginning on PID (post-inoculation day) 3 for 18 days, the magnitude of body weight loss decreased and food intake increased, despite a persistent fever. The treated guinea pigs also survived significantly longer than infected animals receiving placebo injection (21 vs 14 days). Using guinea pig ileum bioassay and radioimmunoassay, we detected significant levels of plasma leukotrienes in Pichinde virus-infected guinea pigs on PID 11 and possibly PID 14. These findings strongly suggest that leukotrienes play a role in the pathogenesis of arenavirus infection and may account in part for the observed cardiac depression, pulmonary edema, and eventual death. PMID- 3025894 TI - Immunological challenge with virus initiates leukotriene C4 production in the heart and induces cardiomyolysis in guinea pigs. AB - Cardiac myolysis was observed in guinea pigs sensitized with vesicular stomatitis virus (VSV), following challenge with this antigen. The phenomenon developed within 1 h of challenge, appearing as islands in the myocardium. The speed and focal nature of the damage point to obstruction of blood flow as a cause of the myolysis. The myolysis was not a toxic effect of the virus itself, but probably a consequence of cardiac anaphylaxis. It occurred only after challenge, and was abolished in 71% of the animals by pretreatment with a mixture of the lipoxygenase-cyclooxygenase inhibitor, BW755C and H1 histamine receptor antagonist, diphenhydramine. Treatment with BW755C alone before challenge prevented myolysis from developing in 46% of the animals. Challenge in vitro with VSV to the perfused, spontaneously beating, sensitized isolated guinea pig heart increased sulfidopeptide-leukotriene (LTC4, LTD4, LTE4) production from undetectable levels (0.5 ng LTD4-equivalent/heart/15' to 13 ng LTD4 equivalent/heart/15'. At the same time, there were derangements in cardiac rate, contractility and coronary outflow typical of cardiac anaphylaxis. The reduction in coronary outflow rate during cardiac anaphylaxis is due largely to the powerful vasoconstrictor effect of LT, as well as perhaps platelet-activating factor. Thus it is speculated that there is a causal relationship between LT release, vasoconstriction, ischemia and myolysis in the heart, following VSV challenge to sensitized guinea pigs. PMID- 3025895 TI - Antiviral activities of gossypol and its derivatives against herpes simplex virus type II. AB - Gossypol, a disequiterpene obtained from cottonseed oil, and a series of peri acylated gossylic nitriles were compared for their antiviral activities against HSV-II and for their toxicities to the host Vero cells. All of the peri-acylated gossylic nitriles exhibited lower cytotoxicities to the host cell than did the parent compound gossypol. Both gossypol and the series of derivatives exhibited antiviral activities against HSV-II when the virus was treated with drug at concentrations as low as 5 X 10(-7) M. Two of the derivatives, gossylic nitrile 1,1'-diacetate and gossylic nitrile-1,1'-divalerate, were capable of inhibiting viral multiplication in Vero cells that were infected with virus before administration of the drug. The results of this study indicate that modification of the aldehyde functional groups on gossypol lowers the toxicity of this drug but does not abolish its antiviral properties. Derivatives of gossypol may be useful antiviral agents. PMID- 3025896 TI - Receptors mediating contraction of isolated human vas deferens. AB - A large body of evidences has suggested the role of adrenergic, opioidergic and other peptidergic receptors in the mediation of animal vas deferens motility. Different animal species showed different neurochemical patterns, so it is to be expected that human vas deferens has its own specific response to several substances, in relation to its peculiar function. In this study we report on the effects of monoaminergic (norepinephrine, dopamine, serotonin, isoproterenol, cholinomimetic drugs) and opioidergic (morphine, buprenorphin, beta-endorphin, met-enkephalin and dynorphin) agonists on isolated human vas deferens motility. Norepinephrine and dopamine provoked complex patterns of motility while opioids did not affect the field electroinduced contractions. The implications of this finding are discussed in relation to human vas deferens function. PMID- 3025897 TI - Effects of beta-casomorphin on 3H-ouabain binding to guinea pig heart membranes. PMID- 3025898 TI - Pharmacology of airway secretion. PMID- 3025899 TI - The benzodiazepine (+)-tifluadom (KC-6128), but not its optical isomer (KC-5911) induces opioid kappa receptor-related EEG power spectra and evoked potential changes. AB - Tifluadom, a benzodiazepine with purported opioid receptor-related analgesic properties, was studied in regard to its acute effects on power spectra of the EEG to demonstrate vigilance changes. Additionally, somatosensory-evoked potentials (SEP) were derived to evaluate its effect on the propagation of impulses in sensory nerve fibers. In order to demonstrate stereospecificity, the two enantiomers of tifluadom (KC-5911 and KC-6128) were given in graded doses (20, 40, 80, 160 micrograms/kg i.v.) to awake, unrestrained and trained dogs at 10-min intervals. KC-6128, but not its optical counterpart KC-5911, induced synchronization of the EEG at 20 micrograms/kg with an increase of power (pW) in the delta (1-4 Hz; +300%), theta (4-8 Hz; +450%), and alpha (8-13 Hz +90%) bands. This was accompanied by a reduction of power in the fast beta domain (13-30 Hz; 95%). Vigilance changes were reflected in the beta/delta quotient which dropped from 3.7 (control) to 0.8 (20 micrograms/kg) and to 0.3 (40 micrograms/kg). A further increase in the dose resulted in saturation. At the highest dose (160 micrograms/kg) there was an additional reduction of the beta/delta quotient to 0.1. In order to unmask the receptor population, possibly mediating the observed changes, a benzodiazepine antagonist and opioid antagonists were given. Ro 15 1788 (240 micrograms/kg) had no effect and naloxone (20 micrograms/kg) induced a short term (5 min) arousal. Only the kappa antagonist Mr 2266 (20 micrograms/kg) induced a reversal of the beta/delta quotient back to 5.5. KC-5911 induced an insignificant drop in the beta/delta quotient which was reversed by Ro 15 1788.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3025901 TI - Transport ATPases in the erythrocytes of rats acclimatized to intermittent altitude hypoxia. AB - The activity of Na+/K+- and Ca2+-ATPase and some allosteric properties of Na+/K+ ATPase were studied in whole erythrocytes and their membrane preparations (ghosts) from rats exposed to intermittent altitude hypoxia (10 and 24 exposures, 8 h/day in an altitude chamber, stepwise up to an altitude of 7,000 m). Ca2+ ATPase activity was increased both in whole erythrocytes and ghosts after the first phase of acclimatization (10 exposures). In a standard incubation medium (containing 3 mmol.l-1 MgCl2 ), Na+/K+-ATPase activity in the ghosts was also increased after the initial phase of acclimatization whereas in whole erythrocytes Na+/K+-ATPase was only decreased in the regression phase. At high MgCl2 concentrations (12 mmol.l-1) changes of Na+/K+-ATPase activity both in whole erythrocytes and in the ghosts followed similar time course with a pronounced increase in the first phase of acclimatization (10 exposures) followed by an abrupt drop (24 exposures) and then by a gradual normalization in the regression phase. Sensitivity of the enzyme to mounting MgCl2 concentrations was increased in the ghosts at the end of acclimatization and was decreased in whole erythrocytes during acclimatization and especially in the regression phase. It has been suggested that chronic altitude hypoxia leads to the alteration of cooperative interaction of the Na+/K+-ATPase subunits in the erythrocyte membrane and accumulation of some factor in the cells inhibiting this enzyme. PMID- 3025900 TI - Mechanisms of tetraethylammonium-induced contraction in the canine coronary artery. AB - Tetraethylammonium chloride (TEA) at concentrations over 10(-3) mol/l produced a concentration-dependent contraction in the isolated canine coronary artery. We investigated mechanisms of the contractile response and the effects of various vasodilators on the contraction. While phentolamine, propranolol, guanethidine, atropine and tetrodotoxin did not affect the TEA response, the response was attenuated by a reduction in the extracellular potassium concentration and was augmented by an increase in the concentration of potassium. The response was also augmented by other potassium conductance blockers (CsCl and 4-AP). On the other hand, the contractile response to phenylephrine used as a control drug was not affected by the extracellular potassium concentration or by CsCl. Verapamil or removal of extracellular calcium abolished the TEA, but not the phenylephrine response. Nicorandil, isoproterenol, adenosine or glyceroltrinitrate produced an equipotent relaxation in the coronary arteries contracted by TEA and by phenylephrine, whereas acetylcholine relaxed to a lesser extent the arteries contracted by TEA than those contracted by phenylephrine. These results suggest that the TEA-induced contractions are strictly dependent on the reduction of potassium conductance and extracellular calcium while the phenylephrine-induced contractions are not and that the relaxing effects of tested vasodilators, except for acetylcholine, may not be affected by such different contractile mechanisms. PMID- 3025902 TI - Opioid elements in development of the spontaneous motility of chick embryos. AB - The effects of the acute and chronic administration of a pure opioid antagonist- naltrexone--was studied in chick embryos from the 4th to the 19th day of incubation. In acute administration, naltrexone (40 mg/kg egg weight) induced paroxysmal activation of spontaneous motility in both normal and spinal embryos from the 13th-15th day of incubation. Activation attained 3- to 4-fold the resting activity of chick embryos of the same ages. The chronic administration of naltrexone (7.46 +/- 1.18 mg/kg e.w. per 24 h) from the 4th to the 16th day of incubation was not manifested either in the embryos' somatic development or in the weight of the brain hemispheres, but it depressed the development of spontaneous motility to 26.1-75.8% of the activity of the control embryos. This developmental effect was not demonstrably correlated either to the length of time for which naltrexone was administered, or to when, in the course of incubation, it was administered to the chick embryos. The results are evaluated as evidence of the participation of opioid elements in the development and effectuation of central motor input functions in the early stages of ontogenetic development. PMID- 3025903 TI - The diagnosis of mononucleosis in the office laboratory. AB - Infectious mononucleosis has been known for a long time to be a common infection in young adults. It also infects children. In particular, children under the age of 2 years may not express the illness clinically. Diagnostic criteria vary but in young adults usually include the symptoms of fever, exudative tonsillitis, and cervical lymphadenopathy. Usually, there is a relative and absolute lymphocytosis, with 10 to 20 per cent or greater atypical lymphocytes. Rapid slide tests are accurate and economical and support the diagnosis when positive. False-positive results are known to occur with several other important disease processes; therefore, the clinical presentation and laboratory results must be interpreted in their appropriate context. Quality control is essential even for the rapid slide tests, and experienced examiners are required to review blood smears. The morphology of atypical lymphocytes varies greatly. In addition, morphology interpretation can be hampered by problems in preparation of slides or the holding of blood samples awaiting slide preparation. EBV-specific serodiagnosis has significantly enhanced our ability to study further and differentiate heterophil-negative syndromes that share clinical features with infectious mononucleosis. Acute, past, chronic, and no EBV infection can now be differentiated. Further diagnostic tests for other etiologic agents are necessary when all EBV tests are negative and the clinical situation requires appropriate diagnosis. PMID- 3025904 TI - Individualized network planning for chronic psychiatric patients. PMID- 3025905 TI - Cushing's syndrome after treatment: changes in cortisol and ACTH levels, and amelioration of the depressive syndrome. AB - Twenty-three patients with pituitary adrenocorticotropic hormone (ACTH)-dependent Cushing's syndrome were studied before and after treatment. The relationship between the amelioration of the depressive syndrome and changes in cortisol and ACTH levels was investigated. There was a significant difference in mean change in 24-hour urinary free cortisol (UFC) excretion for changes in the depressed mood score from first to last visit. There were also significant correlations between decreases in UFC and decreases in both the depressed mood score and the modified Hamilton depression score. These relationships were not found for ACTH. Furthermore, with cortisol decreased to normal levels, continued high ACTH levels did not prevent improvement in depressed mood. The possibility that cortisol may also play a role in the pathogenesis and/or maintenance of the mood disorder in psychiatric patients is discussed. PMID- 3025907 TI - Postmortem serotonergic and adrenergic receptor binding to frontal cortex: correlations with suicide. PMID- 3025906 TI - Beta 2-adrenergic receptor regulation in circulating mononuclear leukocytes in anorexia nervosa and bulimia. AB - Mononuclear leukocyte (MNL) beta 2-adrenergic receptors and their coupled adenylate cyclase system were studied in underweight anorectics (n = 12), weight recovered anorectics (n = 8), bulimics (n = 8), and age- and sex-matched controls (n = 39). Compared with controls, underweight anorectics had significantly fewer MNL beta 2-adrenergic receptor sites (Bmax) but did not differ in binding affinity (Kd). Weight-recovered anorectics and bulimics did not differ from controls on either Bmax or Kd. Compared with controls, all three patient groups had significantly reduced plasma levels of triiodothyronine (T3), while only underweight anorectics had significantly elevated plasma levels of cortisol. Plasma norepinephrine (NE) response to orthostasis was significantly lower in the three patient groups than in controls. The reduction in beta 2-adrenergic receptor number in underweight anorectics could reflect their elevated cortisol and reduced T3 levels. The decrease in beta 2-adrenergic receptor sites, together with the lower NE response to orthostasis, could be responsible for the reduced sympathetic activity of underweight anorectics. PMID- 3025908 TI - Aminergic receptor binding correlates of suicide: methodological issues. PMID- 3025909 TI - Changes in receptor stimulated phosphoinositide hydrolysis in brain during ethanol administration, aging, and other pathological conditions. PMID- 3025910 TI - "Autocannibalism" of membrane choline-phospholipids: physiology and pathology. PMID- 3025911 TI - Effector systems coupled to serotonin receptors in brain: serotonin stimulated phosphoinositide hydrolysis. PMID- 3025912 TI - Serotonin receptor subtype agonists: differential effects on sensorimotor reactivity measured with acoustic startle. PMID- 3025913 TI - Corticotropin-releasing hormone--a new tool to investigate hypothalamic-pituitary adrenocortical physiology in psychiatric patients. PMID- 3025914 TI - Multiendocrine assessment in the dexamethasone suppression test. PMID- 3025915 TI - Lack of serotonergic modulation on 3H-imipramine binding sites in basal conditions and during prolonged treatment with desmethylimipramine. PMID- 3025916 TI - Structure-activity relationships and mechanism of action of hallucinogenic agents based on drug discrimination and radioligand binding studies. PMID- 3025917 TI - Norepinephrine as an 'umbrella' neuromodulator. PMID- 3025918 TI - Effect of 24-hour preservation with oxygen free radical scavengers on isolated perfused canine heart-lungs. AB - The effectiveness of 24-hour hypothermic machine perfusion with TP-V (a hyperosmolar colloid solution containing dextrose, sucrose and ATP-MgCl2) alone, or in combination with oxygen free radical scavengers, was evaluated in isolated perfused canine heart-lungs. Heart-lungs were perfused at 4 degrees C in either TP-V (n = 6), TP-V/Allopurinol (500 mg/L) (n = 6), or TP-V/Allopurinol (500 mg/L) & Catalase (5000 U/L) (n = 5). Lung inflation was maintained with 100% nitrogen. Following preservation, the heart-lungs were perfused with an albumin-mannitol perfusate for 3 hours at 37 degrees C, for functional, hemodynamic, and laboratory determinations. Cold preservation with TP-V/Allopurinol, and TP V/Allopurinol & Catalase resulted in physiologically normal LDH levels during the 3-hour normothermic isolated perfusion test period. Significantly lower enzyme activity for CPK was evident at 0 (p less than .005) and 3 hours (p less than .05) of perfusion, while no significant differences in lactate production were seen among the groups. In addition, pH, PCO2, PO2, and left ventricular, aortic, and coronary artery pressures all remained within normal physiologic range, with no significant differences seen among the three groups. 99m Technetium scans demonstrated adequate patency among the heart-lungs, with better flow seen in those perfused with TP-V/Allopurinol & Catalase. Histological specimens confirmed a decrease in myocardial and pulmonary damage when Allopurinol and/or Catalase was used. It appears that oxygen free radical scavengers provide some protection from canine heart-lungs which have been hypothermically preserved for 24 hours. PMID- 3025920 TI - Anionradicals from 5-halouracil substituted oriented DNA after X-irradiation at low temperatures. PMID- 3025919 TI - Radiation damage to phi X174 DNA and biological effects. AB - Dilute aqueous solutions of biologically active DNA can serve as a simplified model system of the cell. As a biological endpoint the survival of the DNA (after transfection to E. coli spheroplasts) is used. Damage in the DNA, irradiated in water with gamma rays, can be ascribed to reactions with primary waterradicals. By introducing additives in such solutions, which will scavenge the primary waterradicals, competition between a scavenger and DNA for such radicals can be studied. Comparison of different additives makes it possible to decide whether a compound behaves like a simple scavenger, radiosensitizer or like a radioprotector. In this context work has been done with the electron-affinic radiosensitizers metronidazole, misonidazole and nifuroxime. We have found that these wellknown cellular sensitizers do not enhance the inactivation of biologically active DNA. They act as simple competitive scavenger for waterradicals. However, if besides a sensitizer a trace of a metalloporphyrin containing compound (e.g. cyt. c) is present during irradiation an enhanced DNA inactivation, which can be interpreted as sensitization, is observed. Without sensitizer metalloporphyrins induce an enhanced protection of DNA. Apart from these effects the consequences of both chemical-(sulphydryl) and enzymatic (excision; recombination) repair has been studied. It has been found that sulphydryl compounds are able to react with DNA radicals, modifying the radiation damage in such a way that e.g. breaks are prevented. Further in double-stranded DNA a considerable amount of OH and also H radical damage appeared to be reparable by the excision-repair mechanism. However, post-replication repair had only very small or no effect on the amount of damage. PMID- 3025921 TI - Tibial bowing exacerbated by partial premature epiphyseal closure in sex-linked hypophosphatemic rickets. AB - Two girls with sex-linked hypophosphatemic rickets are described in whom premature fusion of the medial proximal tibial epiphyseal plate exacerbated lower extremity bowing. Early recognition of this complication and appropriate surgical intervention--excision of the bony bridge in the growth plate, or epiphysiodesis of the still open portion of the growth plate--might prevent further shortening or deformity of the affected limb. PMID- 3025922 TI - The ubiquitin pathway for the degradation of intracellular proteins. PMID- 3025923 TI - The interferon genes. PMID- 3025924 TI - The cyclic AMP second messenger system in man: the effects of heredity, hormones, drugs, aluminum, age and disease on signal amplification. AB - The intracellular effects of a number of hormonal signals are mediated by the cyclic AMP second messenger system in man and the ubiquitous distribution of hormone-stimulated adenylate cyclase suggests the importance of this enzyme complex in normal aging and pathophysiological states. Various vectors including heredity, endogenous catecholamines, steroid hormones, and drugs affect the activity of hormone-stimulated adenylate cyclase in man. The effect of heredity was studied using lymphocytes obtained from monozygotic twin pairs and age and sex-matched sib pairs. Only for forskolin-stimulated activity is a significant proportion of individual variance attributable to heredity, suggesting the relative stability of the catalytic subunit. Beta-adrenergic and prostaglandin E 1 activity are "state" characteristics and their activities are controlled by environmental parameters. A significant reduction in isoproterenol-stimulated cyclic AMP accumulation between the menses and luteal phase of the menstrual cycle is observed in lymphocytes obtained from 11 female subjects. The lowest level of beta-adrenergic receptor activity is associated with the highest levels of progesterone and estradiol hormone levels in blood. Lithium at therapeutic concentrations markedly inhibits adenylate cyclase activity in platelet membranes. Moreover, marked individual differences are observed in sensitivity to lithium as determined by Dixon plot derived Ki values for 9 normal, healthy subjects. Human adenylate cyclase obtained from platelets and lymphocytes is activated by micromolar amounts of aluminum in the presence of NaF. Irreversible activation of adenylate cyclase by aluminum is suggested as a possible mechanism of this metal's neurotoxicity. The biochemical basis for the age-associated decline in beta-adrenergic responsiveness in man is discussed. Several investigations suggest a deficit at two levels in the adenylate cyclase complex: an impaired coupling of the receptor/N protein subunits and an additional lesion distal to the receptor at the level of N/C coupling. Perfusion studies with salbutamol suggest that the decline in beta-adrenergic sensitivity is general and not restricted to lymphocytes. Possible abnormalities in cyclic AMP signal amplification and recognition in various disease states is discussed. Increased prostaglandin E-1-stimulated cyclic AMP accumulation is observed in lymphocytes obtained from patients with Alzheimer's disease compared to age-matched controls and correlated with severity of the disease state.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3025925 TI - Melanocortins, neural plasticity and aging. AB - Peptides derived from ACHT and alpha-MSH are known to exert trophic influences on peripheral and central nervous structures. Age-related brain diseases may in part be related to loss of neural plasticity. Melanocortins improve adaptional abilities of the nervous system. Chronic treatment with melanocortins may counteract age-related brain pathology. PMID- 3025926 TI - Drug therapy of Alzheimer's disease: realistic or not? AB - Drug therapy of Alzheimer's disease is either symptomatic, directed against behavioral manifestations, or specific, presumably directed against some hypothetical pathogenetic mechanism. Symptomatic treatment involves the use of antipsychotic drugs, alone or combined with benzodiazepines, to curb disturbed behavior, restore a normal sleep-wake cycle and improve self-care. Antidepressants can be used cautiously when indicated. Specific treatment has employed many different drugs over the years, including stimulants, cerebral vasodilators, drugs for certain hypothesized causes of the disorder, and drugs assumed to enhance metabolism of the brain, or to protect it against various insults. Of the metabolic enhancing drugs, ergoloid mesylates, nafronyl and piracetam are of the greatest current interest. Ergoloid mesylates have been the most extensively studied, but many questions about this drug, such as its mechanism of action, its proper dose, the proper duration of treatment, and the proper goals of treatment, remain unanswered. Symptomatic treatment is essential, both of the sake of patients and those who attend them. Specific treatment is more controversial. If specific treatment is warranted, ergoloid mesylates might be a reasonable first-choice. A number of experimental approaches aimed at enhancing deficient neurotransmission, or remedying deficient neuropeptides, are currently being investigated. Meanwhile, it is realistic to attempt to do the best one can for the afflicted patients. PMID- 3025927 TI - Neuropeptides: ACTH-peptides in dementia. AB - Studies on the treatment of demented patients with ACTH neuropeptides are reviewed. The discussion is restricted to ACTH4-10 and an orally active peptide, the modified ACTH4-9 fragment: H-Met (O2)-Glu-His-Phe-D-Lys-Phe-O (Org 2766). The first part of the review concentrates on data from animal pharmacology and basic and human studies which have relevance for the clinical effects shown in demented patients. The second part concerns clinical studies in man. Own results from a placebo-controlled dose finding study on the treatment of 156 PDSD patients with Org 2766 are presented. Org 2766 was in this study found to have a significant therapeutic effect in patients with PDSD measured mainly on the basis of SCAG total, although the effect was found to be rather small (average 4 points improvement on SCAG total). Post hoc analyses demonstrated that effects were clinically relevant for only 25 per cent of the patients. Analyses of SCAG factor scores indicated an effect on cognitive function and somatic function. The study failed to show clear dose-response relationship. However, before a final conclusion on the efficacy of Org 2766 can be reached the results must be confirmed by others. PMID- 3025929 TI - [Type I topoisomerases]. PMID- 3025928 TI - Design and interpretation of opiate antagonist trials in dementia. AB - In view of the reports of possible beneficial effects of naloxone in dementia, rationales and strategies for studying endogenous opiate systems are reviewed. Important considerations in the design and interpretation of clinical investigations using naloxone are also reviewed. The nature and distribution of endogenous opiate systems are summarized from an historical perspective. Endogenous opiate systems are distributed throughout the central nervous system and play important roles in a variety of brain functions, including memory and learning. In view of this, several rationales are evident for studying endogenous opiate systems in dementia, since it is a syndrome in which structures known to contain opiate systems are disturbed, functions modulated by opiate systems are disturbed, and other neurotransmitter systems (functionally linked to endogenous opiate systems) are disturbed. Different strategies for studying endogenous opiate systems are reviewed, including examination of body fluids and pharmacologic challenge studies. Naloxone hydrochloride, a competitive opiate receptor antagonist, is a commonly used pharmacologic agent. The design of a multidose naloxone study of 12 dementia patients is discussed, with reference to the pharmacokinetics, pharmacodynamics, and specificity of naloxone as well as to the nature of the dependent measures selected for this study. No cognitive benefit was observed in this study. Behavioral arousal was observed at naloxone doses, with more evident psychomotor retardation at higher doses. These findings are contrasted with the results of naloxone challenges in other studies. The varying effects of naloxone within and across populations can be conceptualized in terms of the basic and clinical considerations previously discussed. The importance of dose-finding studies is stressed for this and other drug trials. PMID- 3025930 TI - Effects of highly purified eicosapentaenoic acid on vascular reactivity to angiotensin II and norepinephrine in the rabbit. AB - There is disagreement about whether supplementation of the diet with fish oil, which is rich in eicosapentaenoic acid (EPA), lowers blood pressure. We gave highly purified EPA in a soft capsule (90% ethyl ester form of EPA; EPA-E), to female rabbits (100 mg/kg/day) for 4 weeks. Vascular response to vasoconstrictor agents was assessed serially by measuring the systolic blood pressure using a Grand-Rothschild capsule in the ear. There was no change in systolic blood pressure of rabbits treated with EPA-E, but rabbits given EPA-E for one week or longer were significantly less responsive to the pressor effects of angiotensin II than the controls. Responses to norepinephrine did not change. Rabbits given EPA-E for four weeks had significantly more EPA in the serum, but there were no differences in serum levels of triglycerides, total cholesterol, or high-density lipoprotein cholesterol. These results suggest that vascular responses to exogenous angiotensin II can be selectively depressed by short-term treatment with EPA-E in rabbits without changing systolic blood pressure. PMID- 3025931 TI - Pharmacology of peptide leukotrienes on ferret isolated airway smooth muscle. AB - The contractile activities of peptide leukotrienes (LT) on isolated spiral strips of ferret trachea were characterized pharmacologically. LTC4 and LTD4 contracted ferret tracheal strips in a concentration-related manner and were 3- to 8-fold more potent than carbachol. In contrast, high concentrations of LTE4 evoked either weak contractions or none at all, whereas LTC4 and D4 were partial agonists compared to carbachol. In tissues which were unresponsive to LTE4, this compound antagonized contractile responses to LTC4 and D4 in an apparently competitive manner: Carbachol-induced contractions were not altered by LTE4. The cyclooxygenase inhibitor, indomethacin (5 microM), LT antagonist, FPL55712 (10 microM), atropine (1 microM), phenoxybenzamine (10 microM), and LTB4 (10 microM) failed to alter LTC4 and D4 concentration-response curves. The results indicate that ferret trachea is sensitive to the contractile activity of LTC4 and LTD4 but not LTE4. The LT-induced contractions appear to be mediated by a direct action of the LT rather than indirectly through release of secondary mediators such as thromboxane, prostaglandin, or acetylcholine. LT receptors in ferret trachea are insensitive to FPL55712 but are antagonized by LTE4. PMID- 3025932 TI - The prostacyclin/thromboxane balance is favourably shifted in Greenland Eskimos. AB - The rare incidence of cardiovascular disease in Eskimos has been ascribed to their diet rich in eicosapentaenoic acid (EPA, C20:5n-3) and hence a possible formation of trienoic prostanoids. In this study we compare endogenous formation of prostacyclin (PGI), which is formed by the endothelial cell, and thromboxane (TXA), which is formed by platelets in 20 Eskimos and 20 age and sex matched Danish controls by measurement of the main urinary metabolites. Considerable formation of bioactive PGI3 from dietary EPA was shown in Eskimos, which was barely detectable in the controls. Furthermore synthesis of PGI2 was significantly higher in Eskimos in spite of a markedly lower arachidonate content in membrane lipids. In contrast formation of TXA2,3 was lower in Eskimos as compared to the Danish controls. We conclude, that the balance between PGI and TXA, which may regulate the interaction of platelet and vessel wall, is favourably shifted in Greenland Eskimos to an antithrombotic state. PMID- 3025933 TI - Prostaglandin production in cultured gastric mucosal cells: role of cAMP on its modulation. AB - The effect of cAMP on prostaglandin production may depend on cell types. To clarify the relationship between PG and cAMP, we examined arachidonate's effects on PG synthesis and intracellular cAMP accumulation in monolayers of rat gastric mucosal cells. These cells produced PGE2, PGI2 and thromboxaneA2 (TXA2) in amounts of 316 +/- 18, 100 +/- 7 and 30 +/- 5 pg per 10(5) cells in 10 min, respectively, in response to 10 microM arachidonic acid (AA). The production of these PG, however, leveled off subsequently. Cells initially exposed to AA responded poorly to a subsequent stimulation by AA. AA simultaneously stimulated intracellular cAMP accumulation; this stimulatory effect on cAMP production was abolished by the pretreatment with indomethacin. Nevertheless, the pretreatments with dibutyryl cAMP (0.1-5 mM) did not alter the amount of subsequent AA-induced PGE2 production. Furthermore, the preincubation with 1mM isobutyl methyl xanthine also failed to affect PGE2 synthesis, while it increased intracellular cAMP accumulation. Our studies suggest AA stimulates intracellular cAMP formation in cultured gastric mucosal cells, linked with conversion of AA to cyclooxygenase metabolites, AA-induced PG production is limited in these cells, and it seems, however, unlikely that intracellular cAMP modulates AA metabolism to PG. PMID- 3025934 TI - Workshop: Potential therapeutic uses of inhibitors of leukotriene generation and function. June 23, 1986, Bethesda, Maryland. Sponsored by the National Institute of Allergy and Infectious Diseases and the World Health Organization Collaborating Center for Allergic Diseases. PMID- 3025935 TI - Suppression of prostaglandin synthesis by analogues of cyclic AMP and forskolin in chondrocyte monolayer cultures. PMID- 3025936 TI - Leukotriene B3, leukotriene B4 and leukotriene B5; binding to leukotriene B4 receptors on rat and human leukocyte membranes. AB - Specific high-affinity binding sites for [3H]-leukotriene B4 have been identified on membrane preparations from rat and human leukocytes. The rat and human leukocyte membrane preparations show linearity of binding with increasing protein concentration, saturable binding and rapid dissociation of binding by excess unlabelled leukotriene B4. Dissociation constants of 0.5 to 2.5 nM and maximum binding of 5000 fmoles/mg protein were obtained for [3H] leukotriene B4 binding to these preparations. Displacement of [3H]-leukotriene B4 by leukotriene B4 was compared with displacement by leukotriene B3 and leukotriene B5 which differ from leukotriene B4 only by the absence of a double bond at carbon 14 or the presence of an additional double bond at carbon 17, respectively. Leukotriene B3 was shown to be equipotent to leukotriene B4 in ability to displace [3H]-leukotriene B4 from both rat and human leukocyte membranes while leukotriene B5 was 20-50 fold less potent. The relative potencies for the displacement of [3H]-leukotriene B4 by leukotrienes B3, B4 and B5 on rat and human leukocyte membranes were shown to correlate well with their potencies for the induction of the aggregation of rat leukocytes and the chemokinesis of human leukocytes. PMID- 3025937 TI - Sex differences in gastric mucosal protection after 16, 16-dimethyl PGE2 and lithium chloride. AB - While the incidence of duodenal ulcer disease has been documented to be greater in men than in women, this observation has not been previously noted in animal studies of the upper gastrointestinal tract. In this study, we questioned whether the cytoprotective properties of 16, 16-dimethyl PGE2 were sex-related by comparing the degree of ethanol-induced hemorrhagic gastritis in male and female rats pretreated with 16,16-dimethyl PGE2 or lithium chloride. Animals receiving 16,16-dimethyl PGE2 or lithium chloride had significantly less ethanol-induced hemorrhagic gastritis (1.17 +/- 0.15 and 1.24 +/- 0.13, respectively, p less than 0.001) when compared with controls (2.69 +/- 0.10). Female rats treated with 16,16-dimethyl PGE2 had 59% less hemorrhagic gastritis than male rats treated similarly (0.76 +/- 0.14 vs 1.86 +/- 0.19 respectively, p less than 0.001). This sex-related difference in hemorrhagic gastritis was not noted in male and female rats receiving lithium chloride (1.24 +/- 0.15 vs 1.23 +/- 0.27, respectively). However, female rats treated with 16, 16-dimethyl PGE2 had significantly less hemorrhagic gastritis when compared with female rats receiving lithium chloride (0.76 +/- 0.14 vs 1.24 +/- 0.15 respectively, p less than 0.05). PMID- 3025938 TI - Questionable role of leukotriene B4 in monosodium urate (MSU)-induced synovitis in the dog. AB - Monosodium urate (MSU)-induced synovitis in the dog's stifle (knee joint) is similar to an acute gouty attack in man in which a loss of function of the joint correlates with massive influx of neutrophils and the release of an assortment of inflammatory mediators (e.g. histamine, bradykinin, lysosomal enzymes, complement and eicosanoids) into the synovial space. We found in the urate-induced inflammatory exudates 3 hr post MSU the following: 88 million leukocytes/ml (approximately 95% neutrophils) and eicosanoid concentrations of LTB4, LTC4, and PGE2 of less than 0.1, 1.4 and 20 ng/ml, respectively. Isotonic saline injected knee joints at 3 hr contained 5 million leukocytes/ml (approximately 95% neutrophils) and concentrations of LTB4, LTC4, and PGE2 of less than 0.1, 0.7 and 0.2 ng/ml, respectively. Intrasynovial injections of 1 microgram LTB4, 10 micrograms PGE2 or the combination of LTB4 and PGE2 produced no reduction of paw pressure for up to 3 hr. Leukocyte concentrations measured at 3 hr in joints injected with these arachidonic acids metabolites were similar to saline controls. These results question the role of LTB4 as a chemotactic and inflammatory mediator in urate-induced synovitis in the dog but confirm the importance of PGE2 and possibly LTC4 in this model. PMID- 3025939 TI - Production of leukotrienes and prostaglandins in the rat uterus during peri implantation period. AB - We have measured by radioimmunoassay the production of leukotrienes (LTC4 and LTB4) and prostaglandins (PGE2 and PGF2 alpha) in the rat uterus on Days 1 through 6 of pregnancy. The production is defined as the synthesis minus the degradation for a defined period. The production of LTC4 or LTB4 remained unaltered on days 1-3, but exhibited a marked increase on Day 4 showing a peak at noon. This was then followed by a sharp decline on Day-5 morning. A small but consistent peak in uterine LT production was also noticed on Day-5 noon prior to implantation and this was followed by a decline on Day-6 morning i.e. after initiation of implantation. The production profile of PGE2 and PGF2 alpha showed a striking resemblance to that of LTs; one exception being that maximal PG production was noticed on Day-4 morning and preceded the peak production of LTs. These vasoactive arachidonate derivatives reached their peak production rates at around the time when a surge in estrogen level is noticed in the uterus on Day 4. Implantation is a local proinflammatory type of reaction that is associated with increased uterine vascular permeability. Vascular changes in inflammatory reactions are provoked by two kinds of chemical mediators: vasodilators and agents that increase vascular permeability. PGs (especially of the E series) are known as vasodilators, while LTs and histamine mediate increases in vascular permeability. Therefore, an interaction between LTs, PGs, and histamine could be important for uterine preparation for implantation and/or implantation per se. PMID- 3025940 TI - 31P-NMR spectroscopy of myopathies: clinical application of whole-body MR. AB - Patients with myopathies were examined by 31P-NMR spectroscopy using a whole body MR system operating at 1.5 T. Spectra from the femoral muscles were measured as a function of time during a post-exercise recovery period. A 4-min scanning time was used to obtain a good S/N ratio and enable us to evaluate the relative intensity and position of each absorption line. The results were expressed by PCr/Pi ratio and pH value and compared with values before exercise. Clear changes were observed in the recovery process after workload depending on the stage of disease. We were able to observe the in vivo dynamic energy metabolism of myopathies by whole-body MR. PMID- 3025942 TI - Localization of atrial natriuretic peptide, ANP-(99-126) binding sites in the rat thymus and spleen with quantitative autoradiography. AB - Binding sites for atrial natriuretic peptide, ANP-(99-126) were studied in lymphoid organs of the rat with quantitative autoradiography. Tissue sections were incubated in the presence of 0.13 nM 125I-ANP-(99-126) followed by autoradiography using [3H]-Ultrofilm, and the results were analyzed by computerized densitometry and comparison to 125I-standards. Specific ANP binding sites were localized in the medulla and the cortex of the rat thymus and in the white pulp of the rat spleen, with apparent binding sites concentrations of 93, 65, and 126 fmol/mg protein, respectively. The presence of ANP binding sites in areas related to the maturation and function of lymphocytes, and to the production of thymic hormones, suggests the possibility of a role of circulating ANP in the modulation of the immune response. PMID- 3025941 TI - Dynamic MRI of hepatic tumors using gadolinium-DTPA--preliminary results. AB - Dynamic magnetic resonance imaging (MRI) was performed in two patients with hepatic tumors (one cavernous hemangioma and one hepatoma). A sequential timed scanning technique with spin echo images (TR: 100 msec; TE: 20 msec; data acquisition time: 26 sec) was used with a bolus injection of 0.05 mmol/kg gadolinium DTPA. Dynamic MRI findings in the two cases were similar to those obtained with dynamic CT and angiography using iodinated contrast medium. Our early experience suggested that the less invasive technique of dynamic MRI could replace conventional dynamic CT in assessing the hemodynamics of hepatic tumors. PMID- 3025943 TI - [Treatment of microcytic carcinoma of the lung with vincristine, adriamycin, cyclophosphamide and VP-16]. PMID- 3025944 TI - [The cyclic AMP, insulin and glucose responses to glucagon stimulation in patients with liver cirrhosis according to the stage of development ]. PMID- 3025945 TI - [Brain involvement in acquired immunodeficiency syndrome (AIDS): computed tomography (CT) and magnetic resonance tomography (MR)]. AB - Involvement of the central nervous system in acquired immune deficiency syndrome (AIDS) is usually due to opportunistic infections; these frequently offer a difficult differential diagnostic problem. Imaging methods play an important part in the elucidation of symptoms. CT and MR findings were analysed in 13 patients with AIDS and neurological symptoms. Some infections of the central nervous system (encephalitis of unknown aetiology, cytomegalic encephalitis, meningitis) may show cerebral atrophy or even no morphological changes. Toxoplasmosis and PML are the most common opportunistic infections typical changes on CT and MR may lead to diagnosis. MR offers advantages compared with CT in its higher sensitivity for the demonstration even of small lesions. PMID- 3025946 TI - [Magnetic resonance tomography in suspected acoustic neurinoma: technic and differential diagnosis]. AB - Sixty-five patients with suspected acoustic neuromas were examined by CT and MR; the optimal diagnostic procedures are discussed. Extrameatal acoustic neuromas (11 cases) were all diagnosed by CT and MR, but only 82% of intrameatal tumours could be diagnosed by CT combined with air cisternography. Using special surface coils and the paramagnetic contrast medium Gd-DTPA, the accuracy of MR was 100%. All tumours greater than 4 mm were demonstrated. Following clinical and neurological examination, MR is the primary diagnostic method in the investigation of acoustic neuromas. PMID- 3025947 TI - [Postoperative follow-up after thromboendarterectomy of the carotid artery using duplex sonography]. AB - Fifty-one patients, on whom a total of 64 carotid thromboendarterectomies had been performed, were followed up by duplex sonography. Five asymptomatic occlusions of the internal carotid artery (8%) and one occlusion of the common carotid artery were found. Two patients (3%) developed a restenosis of more than 50% and another two patients a restenosis of less than 50%. Changes in the vessel wall not producing significant luminal change were observed in another 18 cases (28%). Thirty-six of the operated vessels (56%) showed no change. PMID- 3025948 TI - [Stress myelography. A new functional study technic in diseases of the lumbar spinal canal]. AB - To optimize functional diagnostics in lumbar syndromes a new myelographic technique was developed termed "loading myelography". During the procedure the patient stands with a 10 kg. weight on his outstretched arms. Based on the law of leverage the load exercised on the vertebral column is more than two and a half times of one-half of the body weight. The author tested the efficacy of the method in 119 patients suffering from disc prolapse, spinal canal stenosis, spondylolisthesis or arachnitis. The results of the conventional myelogram compared with myelography under load conditions demonstrate the value of the method: without load the diagnosis would have remained uncertain in 25% and in 18% load myelogram revealed a pathological finding although conventional myelography was normal. We consider as indications for load myelography: Discrepancy between clinical and conventional myelographic findings; clinically expected multisegmental lesions; spinal canal stenosis; and spondylolisthesis. PMID- 3025949 TI - [High-resolution sonography after surgery of cystadenoma lymphomatosum of the parotid gland]. AB - Clinical and sonographic examinations were carried out on 38 patients who had undergone excision of a parotid gland for adenolymphomas and the results were compared. In 23 of these patients (60%) sonography showed further tumours on the same or opposite side, or bilaterally. The incidence of multiple tumours is therefore significantly higher than has been assumed so far. In 76% of patients, sonography showed clinically occult cystadenoma lymphomatosum. Only 14% of tumours smaller than 12 mm were detected clinically. In four patients, tumours larger than 12 mm, but lying deep in the gland were missed on palpation. In view of the frequency of multi-local or bilateral tumours, sonography of both parotids should be performed if there is clinical suspicion of a parotid tumour. PMID- 3025950 TI - [High-resolution computed tomography in the diagnosis of bone-destroying processes of the external ear]. AB - Eleven patients with known malignant tumours of the outer ear and three patients with otitis externa maligna were examined by high resolution CT. CT provided accurate information concerning soft tissue infiltration into the parotid or subtemporal tissues, and of the bony destruction in the mastoid, meatus and tympanic cavity. Absolute differentiation between a malignant tumour and otitis cisterna maligna is not possible, not even by high resolution CT. PMID- 3025951 TI - [Magnetic resonance tomography in injuries of the cervical spine]. AB - Twenty patients who had suffered spinal trauma were examined by magnetic resonance tomography. Fifteen patients with first degree trauma in Erdmann's classification showed no abnormality. Magnetic resonance tomography of the cervical spine appears to be a suitable method for investigating patients with whiplash injuries. It is indicated following severe flexion injuries with subluxations and neurological symptoms, since it is the only method that can demonstrate the spinal cord directly and completely and show the extent of cord compression. For patients with thoracic trauma and rapidly developing neurological symptoms, magnetic resonance tomography is ideal for showing post traumatic syringomyelia. Magnetic resonance tomography following whiplash injuries is recommended if plain films of the cervical spine show any abnormalities, as well as for the investigation of acute or sub-acute neurological abnormalities. The various findings are discussed. PMID- 3025952 TI - [Findings of NMR tomography in the diagnosis of osteomyelitis and arthritis]. AB - The findings on MR tomography in fourteen patients with osteomyelitis and eight patients with arthritis are described. A retrospective analysis showed that MR was superior in demonstrating changes in the bone marrow and spread of infection round the bone. These changes are shown more accurately and with better contrast after the use of gadolinium DTPA. T1 and T2 sequences appear to be necessary. PMID- 3025954 TI - [Rare anomalies of the architecture of the bronchial tree]. AB - Six cases of rare bronchial anomalies are presented (3 complete right-sided hypo arterial bronchial distributions, 1 partial right-sided hypo-arterial bronchial supply of the upper lobe, 2 cases of atresia of the left apico-posterior bronchus). Emphasis is placed on the bronchographic elucidation of the changed bronchial segmental topic if additive or subtractive bronchial anomalies are found endoscopically especially with regard to preoperative aspects. PMID- 3025953 TI - [Gd-DTPA in magnetic resonance diagnosis of chronic myocardial infarct]. AB - 30 patients with myocardial infarction older than three weeks were examined by MRI prior and following intravenous injection of 0.1-0.2 mmol/kg Gd-DTPA, 201TL SPECT and cineventriculography. Gd-DTPA did not cause any significant change (p less than 0.05) of T2-values for infarcted or non-infarcted myocardium. Compared with this, signal intensity for T1-weighted images increased after the application of the contrast agent both for normal (26 +/- 14%) and infarcted myocardium (33 +/- 16%). The intraindividual signal intensity ratio for infarcted and normal myocardium increased from 1.06 +/- 0.07 to 1.12 +/- 0.09 after the injection of Gd-DTPA. The diagnosis of myocardial infarction by visual analysis of signal intensity was not possible in individual cases. Myocardial infarction could only be visualized indirectly by morphological changes such as wall thinning and aneurysm. PMID- 3025955 TI - [Gastric polyps and their relations with gastric carcinoma]. AB - In 31 of 33 patients with polyps of the stomach who underwent gastroscopy and x ray examination the polyps could be diagnosed roentgenologically. 11 patients had multiple polyps, which were in 8 cases hyperplastic polyps, in 3 "other polyps", whereas in 20 patients with solitary polyps 11 cases showed hyperplastic, in 5 cases adenomatous and in 4 cases "other" polyps. After x-ray analysis it can be stated that in our cases the premalignant adenomatous polyp always occurred solitary. PMID- 3025956 TI - [Hypertrophic pyloric stenosis--an indication for sonography]. AB - The pyloric region of 58 infants who presented with vomiting was examined by real time ultrasound. Transverse diameter of the pylorus and single wall thickness were measured. In 18 patients sonographic diagnosis of hypertrophic pyloric stenosis (HPS) was made and subsequently confirmed by surgery. The mean (+/- one standard deviation) of the transverse diameter and of the pyloric wall thickness in patients with HPS was 15 (+/- 1) mm. and 5.5 (+/- 0.7) mm., respectively. Forty infants matched for age and sex distribution without gastrointestinal symptoms served as controls. The measurements for the transverse diameter were 9.6 (+/- 1.8) mm. and for the wall thickness 1.9 (+/- 0.5) mm. Diameter values of patients with HPS were significantly (p less than 0.001) higher than those of the control group and those of the remaining 40 patients without HPS. Real-time sonography proved a reliable tool in the diagnosis of HPS. PMID- 3025957 TI - [Magnetic resonance tomography of focal hepatic lesions using the para-magnetic contrast medium, gadolinium DTPA. First clinical results]. AB - The use of the para-magnetic contrast medium gadolinium DTPA for magnetic resonance tomography of focal lesions in the liver was investigated in 31 patients. Two dosage schedules of the contrast medium (0.1 and 0.2 mmol/kg body weight were used with field strengths of 0.35 and 0.5 Tesla. Using T1 sequences, gadolinium DTPA showed increased signal intensity in the liver and in tumours, but this was significantly more marked in the tumour. On T1 spin-echo sequences, previously iso-intense lesions became visible after administration of contrast. On the other hand, contrast-enhanced lesions were less well seen on inversion recovery sequences because of a reduction in the contrast between tumour and liver tissue. The contrast between tumour and liver tissue was not improved by gadolinium DTPA in comparison with precontrast inversion recovery sequences and T2 spin-echo sequences. The perfusion of intra-hepatic tumours could be elucidated by magnetic resonance tomography after the administration of gadolinium DTPA. PMID- 3025958 TI - [The accuracy of ultrasound in the diagnosis of intra-abdominal abscess formations]. AB - During the last two years 73 patients with a total number of 84 abdominal abscesses have been investigated by ultrasound. Two thirds of the patients (67%) had postoperative abscesses. Spontaneous abscesses occurred in the remaining 33%. Multiple abscesses occurred in the postoperative group only (7 patients). The overall sensitivity in detecting an abscess was 84%. The sensitivity was much better with spontaneous abscesses (92%) than with postoperative abscesses (80%). One third of the patients have also been investigated by computed tomography. The diagnosis was confirmed either by needle aspiration or by operative drainage. Ultrasound was most useful in detecting abscesses in the right and left upper quadrant and in the pelvis, while CT was superior in the mid-abdomen and when dealing with multiple abscesses. PMID- 3025959 TI - [Diagnostic and therapeutic aspects of antegrade ureteropyelography and percutaneous nephropyelostomy in childhood]. AB - In a sample of 18 children aging one day to eighteen years 21 percutaneous nephrostomies and 27 antegrade pyelographies were performed using sonography and fluoroscopy. Thirteen patients had both nephrostomy and pyelography and 5 had only pyelography. Three of 13 patients needed bilateral nephrostomies. The results indicate that the combination of percutaneous nephrostomy and antegrade pyelography is helpful in diagnosis and treatment of various obstructive disorders of the urinary tract especially in the newborn period. PMID- 3025960 TI - [Two-plane angiography of femoropopliteal vascular prostheses with 90 knee flexion]. AB - The technique of two-plane angiography of femoro-popliteal bypasses with 90 degrees knee flexion is described. Shortening, kinking and stenoses of the bypass are clearly demonstrated by this method. PMID- 3025961 TI - [Image exactness in digital subtraction angiography]. AB - The quality of an imaging method intended to show pathological alterations in cases of arteriosclerosis depends to a large extent on the exactness in imaging the surface morphology of the vessel. In digital subtraction angiography (DSA), the spatial resolution capacity is considerably influenced by the concentration of contrast medium. Studies in vascular models showed that in intravenous DSA (IV DSA) the concentrations of contrast medium are not sufficient to depict minor lesions of the vascular walls. Under the conditions of intraarterial DSA (IA DSA), the imaging exactness increases considerably. It must be stated, however, that the precise representation of the actual finding still continually declines towards the edge of the vessel so that minimal lesions may rather be detected in front or rear walls than in the marginal parts of the vessels. This is the field in which digital and conventional angiography differ considerably. In pulsating vessels, imaging quality requires the greatest possible difference between the video signal of the applied contrast medium and the noise of the motion artifact. Whereas in IV-DSA minor alterations of the vascular wall are masked by pulsation movements, the increased concentrations of contrast medium used in IA-DSA are sufficient for detecting e.g. membrane stenoses of 1 mm. breadth. Nonionic contrast media differ from conventional contrast media, among other things, by their increased viscosity. This causes flow artifacts which are less marked if the contrast medium is diluted to values usual in IA-DSA. PMID- 3025962 TI - [Pressure gradients in catheters during digital subtraction angiography]. AB - The F4 and F5 catheters used for digital subtraction angiography are subjected to high pressure gradients. The paper analyses the pressure gradients resulting from contrast media of different viscosities in catheters of varying internal diameters and related to the rate of injection. These are demonstrated diagrammatically. The pressure sequences which occur in F4 and F5 sequences can be demonstrated on an oscillograph. The usual high pressure injections also result in turbulent flow conditions. PMID- 3025963 TI - [Multilevel structure isolation using modules based on the example of thoracic radiography]. AB - Conventional chest radiographs were digitized and processed by multilevel band pass filtering. By this procedure different structural components are isolated according to their local spatial frequencies. In chest radiographs a textural component can be separated from a skeletal component. Evaluation of radiological images is facilitated because changes can be looked for at relevant levels. A reconstruction of the image is possible, using different weighting factors to enhance interesting structural components. PMID- 3025964 TI - [Suprarenal aortic sclerosis: coral-reef arteriosclerosis. Case report and literature review]. PMID- 3025965 TI - [ Colloid cyst of the 3d ventricle visualized by magnetic resonance tomogram (MRT)]. PMID- 3025966 TI - [Skull abnormalities in a patient with Laurence-Moon-Biedl-Bardet syndrome]. PMID- 3025967 TI - [Computed tomography diagnosis of a large pseudoaneurysm of the heart]. PMID- 3025968 TI - [Aneurysms of the left renal vein. An incidental sonographic finding]. PMID- 3025969 TI - [Metastasis of breast carcinoma to the intestinal tract and endometrium]. PMID- 3025970 TI - [Arthrogryposis multiplex congenita with femoral fracture]. PMID- 3025971 TI - [Comparison of various traps for tsetse flies (Diptera, Glossinidae) in the Ivory Coast and the Congo]. PMID- 3025972 TI - [Preliminary data on the comparative efficacy of 3 toxic attractant traps for tsetse flies: the monoconic trap, the biconic trap and the blue-black screen in the Daloa region (Ivory Coast)]. PMID- 3025973 TI - [Note on an enzootic of adenocarcinoma of the pituitary nasal mucosa in Djalonke ewes]. PMID- 3025974 TI - Bluetongue in camels: a serological survey of the one-humped camel (Camelus dromedarius) in the Sudan. PMID- 3025975 TI - [Proctocolectomy with an ileal reservoir and ileo-anal anastomosis for familial polyposis and ulcerative colitis]. PMID- 3025976 TI - [Rare perianal neoplasms]. PMID- 3025977 TI - GABA-A and GABA-B receptors in the cuneate nucleus of the rat in vivo. AB - Electric stimulation of the rat forepaw evokes a negative potential (N-wave) at the ipsilateral cuneate nucleus. The responses of the N-wave to microiontophoretically applied GABA agonists and antagonists have been studied. Applications of GABA-A agonists (3-amino-propanesulfonic acid and muscimol) reduce the amplitude of the N-wave. This effect decreases during prolonged application, suggesting a desensitization of GABA-A receptors. In addition the effect of muscimol is reduced by (-)-bicuculline methiodide. Baclofen (a GABA-B agonist) also depresses the N-wave but its action lasts longer, is less reversible, shows no desensitization and is not blocked by (-)-bicuculline methiodide. The different responses of the N-wave to GABA-A and GABA-B agonists are compatible with the existence of different types of functional receptors for them in the cuneate nucleus of the rat. The receptors activated by muscimol (GABA A) are clearly not the same as the ones activated by baclofen (conceivably GABA B). PMID- 3025978 TI - The effect of guar gum on the acute metabolic response to glyburide. AB - The effect of 5 g guar gum on the acute blood glucose, insulin and C-peptide response to 5 mg glyburide (HB 419) was investigated in 10 healthy volunteers after an overnight fast. The co-administration of guar gum significantly enhanced the insulinogenic and blood glucose lowering effect of glyburide. PMID- 3025979 TI - Rapid changes in central beta-adrenergic receptors after chronic intravenous infusion of antidepressants. AB - The effects of intravenous infusion of desipramine (1, 3, 10, and 60 mg/kg/24 hr), amitriptyline, zimelidine, iprindole (3, 10, 30, 60, and 100 mg/kg/24 hr each), imipramine (10, 30, and 100 mg/kg/24 hr), or U-48,753E (1, 3, 10, and 30 mg/kg/24 hr) on the density of central beta-adrenergic receptors were investigated in Sprague-Dawley rats. A comparative study with oral desipramine (3 mg/kg/24 hr) for 74 hrs was also carried out. Desipramine, amitriptyline, zimelidine, iprindole, imipramine, and U-48,753E reduced the density of beta adrenergic receptors in the cerebral cortex, and the effect seems to be dose dependent. In the oral study, desipramine failed to down-regulate beta-adrenergic receptors. These results indicate that down-regulation of beta-adrenergic receptors can be rapidly achieved by intravenous infusion of drugs. PMID- 3025980 TI - Effect of indapamide on atrial natriuretic polypeptide receptor in spontaneously hypertensive rat kidney. AB - The effect of a hypotensive state on atrial natriuretic polypeptide (ANP) receptors of the kidney treated by indapamide was investigated in spontaneously hypertensive rats (SHRs). SHRs aged 12 weeks were injected intraperitoneally with indapamide (10mg/kg/day) for 10 days and an ANP radiolabeled receptor assay (RRA) was done on the 11th day. The systolic blood pressure of SHRs injected with indapamide (IDP group) was statistically lower than that of SHRs injected with 2% gum Arabic solution (control group). Concerning the RRA of ANP in the SHR kidney, high affinity and low capacity binding sites were observed in the IDP group (Kd = 0.220 +/- 0.059 nM, Bmax = 6.10 +/- 2.36 fmol/100 micrograms) compared with the control group (Kd = 0.401 +/- 0.147 nM, Bmax = 9.96 +/- 2.50 fmol/100 micrograms). These finding suggested that the hypotensive state induced by treatment with indapamide may change the ANP receptor in kidneys of a SHR. PMID- 3025981 TI - DNA binding by adriamycin, melphalan, cis-dichloro-diammineplatinum (II) and nornitrogen mustard assessed by restriction enzyme block. AB - The ability of a number of DNA binding drugs to block the action of selected restriction enzymes has been used to determine the nucleotide base binding preference of these drugs. The three alkylating agents studied all displayed guanine-cytosine (G-C) base binding preference which accords with previous results for cis-DDP and with the preferred sites of alkylation being the N7 and O6 of guanine bases. Adriamycin was markedly more reactive in this assay system compared to the alkylating drugs and in contrast to these drugs showed an adenine thymine (A-T) base binding preference. PMID- 3025982 TI - Arterial ammonemia changes of renal origin induced in the rat by acid and alkaline diets. AB - The aim of this study was to investigate the influence of acid and alkali food supplementation on systemic ammonemia to explain the hyperammonemia previously observed in rats fed a high protein diet. In normal rats, arterial ammonia concentration significantly increases after 4 days of HCl-supplemented diet. Following a NaHCO3-enriched food, there is only a slight but not significant decrease in arterial ammonia level. These changes occur before any variation in arterial acid-base status and are of renal origin. Indeed, there is a positive linear correlation (r = 0.946; P less than 0.001) between arterial ammonia level and the ammonia concentration difference between the renal vein and artery (which varies proportionally to the urinary ammonium excretion). Hindquarter uptake and intestinal release of ammonia do not significantly participate in the arterial ammonia changes observed. Following HCl-enriched diets, increased renal glutamine uptake, enhanced hindquarter glutamine release, and perhaps decreased intestinal glutamine uptake occur simultaneously. In conclusion, acid and alkali food supplementation intervenes on the renal ammonia release into the circulation with concomitant arterial ammonemia changes. PMID- 3025983 TI - Plasma concentrations of angiotensin II and aldosterone during acute left ventricular failure in the dog. Effect of converting enzyme inhibition. AB - Acute left ventricular failure was induced in anesthetized dogs by left coronary embolization. Treatment with the converting enzyme inhibitor enalaprilat (MK-422) was then given. Hemodynamic registrations confirmed the development of left ventricular failure. Plasma concentrations of angiotensin II and aldosterone rose significantly. Treatment with enalaprilat was accompanied by significant reductions in heart preload and afterload and in plasma hormone concentrations. PMID- 3025984 TI - Specific bronchial reactivity to toluene diisocyanate: dose-response relationship. AB - 20 patients with toluene diisocyanate (TDI)-induced asthma were examined in order to assess their threshold of response to TDI during specific bronchial provocative tests (BPT). Specific bronchial hyperresponsiveness was evaluated by performing, on different days, specific BPT with increasing concentrations of TDI until a positive response was obtained; the threshold of response to TDI (low: 0.02-0.05 ppm; moderate: 0.1 ppm; high: 0.2-0.25 ppm) and the pattern of positive response were evaluated in comparison with some clinical features of the disease. The threshold of airway response to TDI was low in 9, moderate in 7 and high in 4 patients. No evident relationship was observed between the threshold of response to TDI and the pattern of positive response to the lower TDI concentration (immediate in 5, late in 8 and dual in 7 subjects) or other clinical features (duration of asthmatic symptoms, smoking habits, cessation of work, nonspecific bronchial hyperresponsiveness to methacholine); however, 6 out of 9 patients with low threshold had nonspecific bronchial hyperreactivity in comparison with 6 out of 11 patients with moderate or high threshold. In 10 out of 13 patients who performed two positive BPT with different TDI concentrations, the pattern of response was the same either at lower and at higher TDI concentrations; 3 subjects who had a late reaction at the lower concentration showed a dual reaction to the higher TDI concentration. A relationship between the degree of the specific bronchial reaction (% fall in FEV1 from baseline value) and TDI concentration during BPT was observed for the immediate reaction but not for the late reaction. PMID- 3025985 TI - Ultrastructure of retinal pigment epithelial and neural cells in the neuronal ceroid-lipofuscinosis affected Dalmatian dog. AB - Ubiquitous accumulation of lipopigments, entailing lamellar, fingerprint, and curvilinear bodies in varying cell types of the retina, and, in addition, peculiar circular inclusions within retinal pigment epithelial cells in the retina of two Dalmatian dogs afflicted with neuronal ceroid-lipofuscinosis, emphasize the identity of its retinal ultrastructural pathology to that observed in NCL-affected English setters, while the retinal layers appear largely preserved. The circular inclusions in RPE cells are unique to canine NCL; they are not encountered in human NCL, nor in primary non-NCL retinal dystrophies of other canine species. Their origin and pathogenesis, so far, remain subject to speculation, rather than to convincing explanation. PMID- 3025986 TI - [Effect of dibutyryl cyclic AMP on hypoxic pulmonary vasoconstriction]. PMID- 3025988 TI - [Protein C]. PMID- 3025987 TI - Influence of lung volume on the mixing defect for an inert gas. AB - For quantitative assessment of how gas mixing in the lung compares to the ideal of perfect mixing, it is necessary to measure pre-inspiratory lung volume as well as to study the inhaled and exhaled gas. Normal young-adult persons in the seated position assumed various lung volumes and then took 1-L test breaths (11% Ar, 22% O2 in N2). Integration of concentration at the mouth against volume yielded amounts of Ar inhaled, exhaled and retained; un-exhaled concentration in the lung after the single breath was calculated from amount retained divided by lung volume (measured by rebreathing). Retention was 60% of the inhaled amount when pre-inspiratory volume was above FRC and less when it was below. Series dead space increased with lung volume. End-tidal Ar was the same as, or slightly above, the concentration predicted for perfect mixing and un-exhaled Ar was slightly below the perfect-mix concentration. INTERPRETATION: three separable influences decrease the transfer of inspirate to the resident gas. A dilution effect, predictable from the volumes involved, decreases retention at low lung volumes. An enlarged series dead space decreases retention at high volumes. In normal persons, a mixing defect that is attributable to poor distribution or incomplete gas-phase diffusion is small and can be ascribed almost completely to the part of the expirate that gives rise to the slope of the alveolar plateau, except when breaths are taken at very low lung volumes. PMID- 3025989 TI - [Neurologic manifestations in systemic vasculitis]. PMID- 3025990 TI - [Opportunistic diseases in patients with HIV infection]. PMID- 3025991 TI - [Right-side abdominal pain, weight loss]. PMID- 3025992 TI - [Painful legs and moving toes. A drug-induced case]. AB - In this case of painful legs and moving toes the essential characteristics were to be secondary to a neuropathy due to vincristine and metronidazole. The disorders regressed in six weeks after the arrest of chemotherapy. To our knowledge, this is the first case due to a toxic, and more precisely to a drug. PMID- 3025993 TI - Immunosuppression in viral infections. AB - Viruses may cause immunosuppression by a variety of mechanisms. This review delineates four categories. First, immunosuppression can result from the direct effects of viral replication on lymphocyte functions. Either all classes of lymphocytes can be affected, as occurs in measles, or the effect can be restricted to a cell subtype, as is the case with human T cell-lymphotropic virus type III. Second, the activity of soluble factors of viral or host origin released from infected cells can affect immunosuppression. A third mechanism results from viral infection of macrophages and affects the function of these cells in natural and acquired immunity. Finally, immunosuppression may result from viral triggering of an imbalance in immune regulation, which culminates in the overactivity of suppressor cells. A detailed knowledge of the mechanisms by which viruses are involved in immunosuppression may help in the design of strategies to reverse the effect. PMID- 3025994 TI - Chemotherapy for Chagas' disease: a perspective of current therapy and considerations for future research. AB - Our current knowledge of the biology of Trypanosoma cruzi and its relation to the development of chemotherapy for Chagas' disease are reviewed. This includes recent developments in the understanding of kinetoplast DNA and glycosomes; the action of oxygen radicals; intermediary metabolism of purines, pyrimidines, and folic acid; and the formation of microtubules. At this time, these organelles and metabolic pathways appear to be the most promising for potential exploitation for chemotherapeutic purposes. Compounds of current experimental interest also are discussed. These are agents that have shown promise in the laboratory and for which data exist regarding probable mechanisms of action. The activities of these agents correlated, in so far as is possible, with those structures or metabolic pathways in the trypanonsome that are affected by their actions. The compounds are of two general groups: nitro compounds and purine analogues. PMID- 3025995 TI - Enzyme-mediated resistance to beta-lactam antibiotics: a symposium on sulbactam/ampicillin. Cairo, Egypt, April 18-19, 1985. PMID- 3025996 TI - Sulbactam: biochemical factors involved in its synergy with ampicillin. AB - Sulbactam is a time-dependent irreversible inhibitor of various beta-lactamases by reversible formation of a Michaelis-type enzyme-inhibitor complex and progressive evolution of this complex into inactivated protein(s). This process is either irreversible (true inactivation) or quasi-irreversible (stable acyl enzyme). In this way, sulbactam efficiently protects ampicillin from degradation by beta-lactamases. Sulbactam itself exhibits a moderate antibacterial activity that is related to an affinity for the penicillin-binding proteins of various bacterial strains, which is similar to the affinity of penicillins such as ampicillin. However, sulbactam binding differs according to the bacterial species involved. In strains producing either low levels of beta-lactamase or none at all, a synergistic effect, minor but not negligible, can be observed when sulbactam is associated with a beta-lactam antibiotic with a complementary affinity for the target sites. PMID- 3025997 TI - Pharmacokinetics of sulbactam/ampicillin in humans: a review. AB - The parenteral kinetics of sulbactam, a potent synergist with ampicillin against a broad range of clinically important organisms in humans, are similar to those of ampicillin. The kinetics of ampicillin were not affected by co-administration of sulbactam, and high levels of both agents were attained: 15-min infusions of 2 g of ampicillin plus 1 g of sulbactam produced peak serum concentrations of approximately 120 micrograms of ampicillin/ml plus 60 micrograms of sulbactam/ml; intramuscular injections of 1 g of ampicillin plus 0.5 g of sulbactam produced peak concentrations of 18 micrograms of ampicillin/ml plus 13 micrograms of sulbactam/ml. The drugs had similar half-lives (approximately 1 hr), and both drugs were excreted primarily in the urine (greater than 75%). Although the kinetics of sulbactam in postpartem women and in surgical patients were similar to the kinetics in young men, the half-life of sulbactam (like that of ampicillin) was altered in the elderly, during labor, in neonates, and in patients with renal impairment. After distribution of the agents in the body, the concentrations of both drugs in blister and parenteral fluid were similar to those in serum. Furthermore, useful antibacterial concentrations of both drugs were found in pus, sputum, and middle-ear fluid. The normally low penetration of sulbactam and ampicillin into cerebrospinal fluid was increased in patients with bacterial meningitis. PMID- 3025998 TI - Single-dose pharmacokinetics of intravenous sulbactam in pediatric patients. AB - The pharmacokinetics of intravenously administered sulbactam were studied in 17 pediatric patients two to 14 years of age. Single doses of 12.5 or 25 mg/kg were infused over 3 min, and in previously healthy children, mean peak plasma concentrations 5 min after dosing were 71 and 163 micrograms/ml, respectively. Noncompartmental and compartmental calculations resulted in similar pharmacokinetic parameters. Linear pharmacokinetics were found in the concentration range studied. The mean terminal-phase half-life was 1.75 hr, the mean total plasma clearance was 180 ml/min per 1.73 m2, and the mean apparent volume of distribution was 340 ml/kg. Approximately 70%-80% of an intravenous dose was excreted unchanged in the urine. In children with cystic fibrosis, both total plasma clearance and apparent volume of distribution were significantly increased. The data support the intravenous administration of 12.5-25 mg of sulbactam/kg every 6 to 8 hr for assessing the adequacy of this drug as an adjunct to beta-lactam therapy for various bacterial infections in children. PMID- 3025999 TI - Pharmacokinetic study of sulbactam and ampicillin administered concomitantly by intraarterial or intravenous infusion in the newborn. AB - The combination of sulbactam and ampicillin was administered to 16 newborn infants, 15 preterm and one term, who required umbilical arterial or venous catheterization and prophylactic antibiotics. The aims were to determine an appropriate dosage regimen and to study the pharmacokinetics. Satisfactory plasma concentrations were achieved with administration of a bolus injection of 50 mg of each drug/kg every 12 hr (mean concentrations: sulbactam, 110 mg/liter; ampicillin, 87 mg/liter 3 hr after dosing; and sulbactam, 105 and 135 mg/liter; ampicillin, 320 and 310 mg/liter 30 min after dosing in two infants. Mean elimination half-lives were longer than those in adults (sulbactam, 7.9 hr; ampicillin, 9.4 hr), and urinary excretion over 12 hr varied considerably (range: sulbactam, 7%-91%; ampicillin, 5%-132%), rates reflecting the immature renal function in the newborn and the relative oliguria characteristic of preterm infants with idiopathic respiratory distress syndrome. There was little evidence of accumulation of either drug, and both were well tolerated. This combination and dosage should be suitable for a trial of therapy for infection in the newborn. PMID- 3026000 TI - Stability of sulbactam/ampicillin in diluents for parenteral administration. AB - Compatibility studies were conducted of sulbactam/ampicillin in infusion diluents that are reported to be compatible with ampicillin sodium. A high-performance liquid chromatographic system that simultaneously detects sulbactam and ampicillin was used to determine whether the infusion diluents and the conditions of use recommended for ampicillin sodium are applicable to sulbactam/ampicillin. The results show that the sulbactam/ampicillin preparation for parenteral administration is compatible with all diluents recommended for ampicillin sodium. In all diluents sulbactam was more stable than ampicillin. At the end of the reported use periods, the average retention value for sulbactam in the combination solutions was 96% (range, 91%-101%), whereas the average retention value for ampicillin in the same solutions was 90% (range, 84%-95%). The presence of sulbactam had no adverse effect on the stability of ampicillin. The average retention value for ampicillin alone in the same diluents was 92% (range, 82% 99%). The conditions of use for sulbactam/ampicillin in diluents for parenteral administration are unrestricted by the presence of sulbactam and are, in fact, governed by those of ampicillin sodium. PMID- 3026001 TI - Sulbactam/ampicillin: in vitro spectrum, potency, and activity in models of acute infection. AB - More than 90% of community hospital-isolated strains of Staphylococcus (including methicillin-resistant isolates), Streptococcus, Haemophilus, Neisseria, Branhamella, Bacteroides, Escherichia coli, Klebsiella, Enterobacter aerogenes, Proteus, and Acinetobacter calcoaceticus were inhibited by the sulbactam/ampicillin (1:2) combination at concentrations of 8 micrograms/16 micrograms per ml. The peak serum level from a 15-min infusion of 1 g/2 g of sulbactam/ampicillin is more than seven times this 90% end point. Excellent bactericidal activity was demonstrated against ampicillin-resistant isolates. Ampicillin-resistant strains did not develop resistance to sulbactam/ampicillin when they were serially transferred in the presence of sublethal concentrations of the combination. In mice the combination was active against a variety of acute, fatal infections produced by ampicillin-resistant bacterial isolates, including methicillin-resistant strains of Staphylococcus aureus and mixed anaerobes. The in vitro and in vivo properties of sulbactam/ampicillin, coupled with its reliable pharmacokinetic performance, appear to make the combination ideally suited for the treatment of polymicrobial (aerobe-anaerobe) infections. PMID- 3026002 TI - Activity of sulbactam/ampicillin in screening and discriminative animal models of infection. AB - The efficacy of sulbactam/ampicillin in the treatment of mice with fatal systemic infections produced by ampicillin-resistant Staphylococcus aureus, Haemophilus influenzae, Klebsiella pneumoniae, or Proteus vulgaris strains is well established. In this paper the demonstrations of efficacy for sulbactam/ampicillin have been extended to a number of clinically relevant models, including bacteremia and meningitis produced by H. influenzae in infant rats, experimental staphylococcal endocarditis in rabbits, localized lesions in mice, urinary tract infections in rats, and prophylaxis in a surgical wound model in mice. In these models, in which ampicillin-resistant organisms were used, sulbactam/ampicillin was either more effective than or as effective as appropriate control agents. Neither sulbactam nor ampicillin used separately displayed significant activity. The results of supportive pharmacokinetic studies, in which differential bioassays were used, demonstrated that sulbactam and ampicillin generally were delivered with equal efficiency to plasma and to extravascular fluids obtained by sampling the contents of implanted cylinders. PMID- 3026003 TI - In vitro bactericidal activity of sulbactam plus ampicillin against methicillin resistant Staphylococcus aureus. AB - By a microtiter checkerboard method, the in vitro bactericidal activity of sulbactam/ampicillin was tested against three groups of Staphylococcus aureus strains: group PR (penicillin-resistant through the production of penicillinase; methicillin-susceptible both at 30 degrees C in medium containing 5% NaCl and at 35 degrees C in medium without NaCl; n = 10); group MR1 (methicillin-resistant under the former conditions but not under the latter; n = 15); and group MR2 (methicillin-resistant under both sets of conditions; n = 20). Sulbactam/ampicillin was synergistic against all strains (fractional bactericidal index, less than or equal to 0.10). Moreover, there was a linear relation in each group between the different concentrations of the two antibiotics required for bactericidal activity. Thus, an equation of bactericidal synergy could be formulated for the combination. The bactericidal activity of sulbactam/ampicillin was assessed against one strain of each group and against a reference strain (S. aureus ATCC 6538 P) in an in vitro model simulating the kinetics of both antibiotics in human blood after intravenous administration of 2 g of ampicillin and 1 g of sulbactam. The observed rate of killing was approximately 99% except with the MR2 strains, against which the combination was bacteriostatic. PMID- 3026004 TI - Induction/inhibition of chromosomal beta-lactamases by beta-lactamase inhibitors. AB - Many species of gram-negative bacteria produce chromosomal beta-lactamases that can be induced to high-level expression by exposure to beta-lactam antibiotics. Fifty-four strains of Escherichia coli, Enterobacter, Proteus, Citrobacter, and Morganella were examined for beta-lactamase inducibility by clavulanic acid, sulbactam, YTR 830, ampicillin, and cefoxitin. Cefoxitin proved to be the most potent inducer, affecting most of the strains of Enterobacter and Morganella. Clavulanic acid induced 30% of all strains studied. Sulbactam and YTR 830 did not induce measurable levels of beta-lactamases. Inhibition of some chromosomal beta lactamases by sulbactam and YTR 830 was demonstrated, whereas inhibitory activity by clavulanic acid was found only in Proteus and Citrobacter. PMID- 3026005 TI - A randomized, double-blind comparison of sulbactam/ampicillin and clindamycin for the treatment of aerobic and aerobic-anaerobic infections. AB - In a randomized, prospective, double-blind trial, sulbactam/ampicillin was compared with clindamycin in terms of efficacy and safety for the treatment of bacterial infections. Both sulbactam/ampicillin and clindamycin were given with gentamicin when this course was indicated by clinical or laboratory findings. In five patients the site of infection was pleuropulmonary; in 14, bone; in 11, skin and soft tissue; and in one, intraabdominal. The commonest anaerobes isolated were anaerobic cocci and Bacteroides species; the commonest aerobic and facultative bacteria were Enterobacteriaceae, Pseudomonas aeruginosa, and various gram-positive cocci. All of six assessable patients given sulbactam/ampicillin alone had satisfactory clinical responses, as did seven of nine patients given sulbactam/ampicillin plus gentamicin, all of six patients given clindamycin alone, and six of nine patients given clindamycin plus gentamicin. Pathogens were totally or partially eradicated in four of five, eight of nine, four of five, and three of nine assessable patients given these same regimens. Adverse reactions and laboratory abnormalities were relatively uncommon. Overall, sulbactam/ampicillin was as effective as clindamycin in the treatment of aerobic or mixed aerobic-anaerobic infections; however, the concomitant use of gentamicin was frequently required with both regimens. PMID- 3026006 TI - Effect of prophylactic administration of sulbactam/ampicillin on the rate of postoperative endometritis after first-trimester abortion. AB - In a single-blind randomized study, the efficacy of a single intravenous dose of sulbactam (0.5 g) plus ampicillin (1 g) was compared with that of placebo in the prophylaxis of early postoperative endometritis after abortion with vacuum aspiration in the first trimester. Administration of either sulbactam/ampicillin or placebo occurred immediately before the operation. The two treatment groups were equally matched with respect to patient's age, gestational age, presence of an intrauterine device, and baseline bacteriologic findings. During the first seven postoperative days, four of 145 patients who had received sulbactam/ampicillin and 11 of 140 patients who had received placebo were diagnosed as having endometritis. No adverse effects and no laboratory test abnormalities attributable to treatment were reported in either group. A single dose of sulbactam/ampicillin reduced the incidence of early postoperative endometritis after abortion with vacuum aspiration in the first trimester. PMID- 3026007 TI - Efficacy of sulbactam plus ampicillin in gynecologic infections. AB - The efficacy of sulbactam plus ampicillin in the treatment of various gynecologic infections was evaluated in 24 women (median age, 35 years). Ten women had pelvic cellulitis plus vaginal cuff abscess; six, pyeloperitonitis; three, vaginal cuff abscess; three, surgical wound sepsis; one, tubo-ovarian abscess; and one, endometritis. Surgical procedures preceding infection included abdominal hysterectomy, ovarian cyst removal, ectopic pregnancy, correction of cystocele, and uterine dilatation and curettage. Twenty patients received 1 g of sulbactam plus 1 g of ampicillin per dose; four received 0.5 g of sulbactam plus 1 g of ampicillin per dose. The combination was given iv every 6 hr for three to four days and then im every 8 hr for three to five days (mean treatment duration, seven days). Pus cultures yielded Enterobacteriaceae (21 cases), enterococci (two), Bacteroides fragilis (12), other Bacteroides species (five), Peptococcus species (nine), Peptostreptococcus species (seven), and other anaerobes (five). Six infections were purely anaerobic; 18 were mixed. All but two infections were cured by both clinical and bacteriologic criteria, with no adverse reactions. Parenteral sulbactam/ampicillin seems safe and effective in the treatment of gynecologic infections of moderate severity. PMID- 3026008 TI - Biliary pharmacokinetics of sulbactam plus ampicillin in humans. AB - The biliary pharmacokinetics of sulbactam and ampicillin was investigated in 19 patients with normal liver function who were undergoing surgery of the biliary tract. The combination of sulbactam (0.5 g) plus ampicillin (1 g) was given intravenously to five patients with T-tube drainage of the common bile duct. Mean peak concentrations of sulbactam (19.4 micrograms/ml) and ampicillin (471 micrograms/ml) in the bile occurred 0.5-1 hr after administration. Biliary excretion was estimated to account for approximately 0.24% of the sulbactam dose and 2.8% of the ampicillin dose. Fourteen other patients received the same dose of sulbactam/ampicillin immediately before elective cholecystectomy. Respective mean concentrations of sulbactam and ampicillin at the time of gallbladder removal were 4.3 micrograms/ml and 15.9 micrograms/ml in gallbladder bile and 6.3 micrograms/g and 7.7 micrograms/g in gallbladder wall tissue. These results, together with the antibacterial spectrum and potency of the combination, suggest that sulbactam/ampicillin is suitable for prophylactic use in biliary tract surgery. PMID- 3026009 TI - An open, comparative study of sulbactam plus ampicillin vs. cefotaxime as initial therapy for serious soft tissue and bone and joint infections. AB - In an open, randomized, comparative study, patients with bone, joint, or soft tissue infections were treated with sulbactam sodium plus ampicillin (sulbactam/ampicillin, 1 g of sulbactam and 2 g of ampicillin three times a day) or with cefotaxime (2 g three times a day) for two weeks as initial therapy. Thirteen patients were entered into the sulbactam/ampicillin group and nine into the cefotaxime group. Two weeks after the end of therapy, clinical cure or improvement was observed in all 13 patients treated with sulbactam/ampicillin and in seven of nine patients treated with cefotaxime. Staphylococcus aureus was not eliminated from one patient in each group. Recurrence of S. aureus infection occurred in two patients in the cefotaxime group within two weeks after the end of therapy. No adverse effects warranting discontinuation of antibiotic therapy were observed. The combination of sulbactam sodium plus ampicillin was at least equivalent to cefotaxime for the initial treatment of acute serious soft tissue and bone and joint infections. PMID- 3026010 TI - Treatment of infections due to multiresistant Neisseria gonorrhoeae with sulbactam/ampicillin. AB - Between January and April 1984, 229 of 448 male patients with urethritis at the Choong-Ku Venereal Disease Clinic in Seoul had positive urethral cultures: 66 for penicillinase-producing Neisseria gonorrhoeae (PPNG) and 163 for non penicillinase-producing N. gonorrhoeae (non-PPNG). Forty-five men with PPNG urethritis were enrolled in a study of the efficacy of treatment with sulbactam/ampicillin plus probenecid. Diagnosis and evaluation of cure were based on culture results. The agar-plate dilution method was used for susceptibility testing, and the chromogenic cephalosporin test was used for detection of beta lactamases. MICs of various antibiotics for the isolates were high. MICs of sulbactam/ampicillin were 0.25-4 micrograms/ml, with an MIC90 of 4 micrograms/ml, a value 16-fold lower than that for ampicillin alone (MIC90 greater than 32 micrograms/ml). Patients were treated with 1 g of probenecid orally and either one vial of sodium sulbactam/ampicillin or two vials intramuscularly. Each vial contained 0.5 g of sodium sulbactam and 1 g of sodium ampicillin. Patients were followed up for three to five days. All patients but one were cured, and no remarkable adverse reactions were noted. The two regimens of sulbactam/ampicillin were equally effective in the treatment of uncomplicated PPNG in men. PMID- 3026011 TI - In vitro activity of aminopenicillins combined with sulbactam, clavulanic acid, or amdinocillin against bacteria isolated from patients with complicated urinary tract infections. AB - The minimal inhibitory concentrations (MICs) of ampicillin, sulbactam/ampicillin, clavulanic acid/amoxicillin, and amdinocillin/ampicillin were determined for 400 bacteria isolated from the urine of 400 urologic inpatients with complicated and/or hospital-acquired urinary tract infections (UTIs). The spectrum of isolates consisted of about one-third each of Escherichia coli, other gram negative strains, and gram-positive strains. About one-third of all isolates were resistant to ampicillin (MICs greater than 8 micrograms/ml). Combination of an aminopenicillin with a beta-lactamase inhibitor improved the activity of the aminopenicillin against gram-negative as well as gram-positive strains. The combination of ampicillin with amdinocillin improved the activity of ampicillin against gram-negative strains only. If the total antibacterial spectrum is considered, the improvements in activity provided by the three combinations were quite similar. PMID- 3026012 TI - Penetration of sulbactam into cerebrospinal fluid of patients with viral meningitis or without meningitis. AB - In a study designed to determine the penetration of sulbactam into the central nervous system, a single intramuscular dose of 0.5-1.0 g was given to 19 patients: nine with viral meningitis and 10 with normal cerebrospinal fluid (CSF). Drug levels in CSF were first detectable 2 hr after administration in patients with viral meningitis and 4 hr after administration in patients with normal CSF. The sulbactam levels were low (0-3.7 mg/ml), and no significant differences were found between levels in the two groups of patients. For the achievement of therapeutic concentrations of sulbactam in CSF, doses larger than those administered in this study should be used in conjunction with a beta-lactam antibiotic. PMID- 3026013 TI - Penetration of sulbactam into the cerebrospinal fluid of patients with bacterial meningitis receiving ampicillin therapy. AB - Concentrations of sulbactam in the CSF of 18 patients with bacterial meningitis who were undergoing treatment with intravenous (iv) ampicillin were determined. Six patients received single doses of sulbactam (1 g) and 12 patients received multiple doses (four times daily) by the iv route at various intervals before lumbar punctures were performed to monitor their condition. Concentrations of sulbactam up to 12 micrograms/ml were detected in the CSF between 1 and 4 hr after dosing, the higher levels being present in the CSF of patients with the most severe meningeal inflammation. There were no significant differences in the concentrations achieved after single or multiple doses of sulbactam, and the concentrations were generally similar to the concurrent concentrations of ampicillin. It is concluded that these results as well as the antibacterial properties of sulbactam plus ampicillin support the evaluation of this combination as an alternative in the treatment of bacterial meningitis. PMID- 3026014 TI - Sulbactam/ampicillin in the treatment of acute epiglottitis in children. AB - Acute epiglottitis, a life-threatening illness, is characterized by the sudden onset and rapid progression of respiratory obstruction. The etiologic agent is almost exclusively Haemophilus influenzae type b (Hib). During the past decade as many as 25% of strains of Hib have been shown to produce beta-lactamase and be resistant to ampicillin. Recommendations for treatment, in addition to the immediate intubation of the airway, include the administration of chloramphenicol in combination with ampicillin. The combination of sulbactam and ampicillin was evaluated in an effort to develop a safer, but equally effective, regimen. Thirty one infants and children (mean age, three years six months) with documented acute epiglottitis received parenteral sulbactam sodium (30 mg/kg per day) in combination with ampicillin (200 mg/kg per day). Of the 31 subjects, 26 (84%) had Hib isolated from the blood; seven (27%) of the 26 strains of Hib isolated were beta-lactamase-positive. Twenty-five cases (96%) of Hib epiglottitis responded rapidly to treatment. The combination of sulbactam and ampicillin appeared to be an effective and safe alternative to chloramphenicol/ampicillin therapy for acute epiglottitis in infants and children. PMID- 3026015 TI - Sulbactam/ampicillin vs. chloramphenicol/ampicillin for the treatment of meningitis in infants and children. AB - Eighty-one patients ages one month to 14 years with meningitis were randomized to receive either sulbactam (50 mg/kg per day) and ampicillin (400 mg/kg per day; 41 patients) or chloramphenicol and ampicillin (40 patients). The groups were comparable in terms of sex and degree of illness; however, more patients treated with chloramphenicol/ampicillin than patients treated with sulbactam/ampicillin were younger than 12 months of age (78% vs. 56%). Pathogens were isolated from the cerebrospinal fluid (CSF) of 65 (80%) of the 81 patients. In the sulbactam/ampicillin group, there were 18 Haemophilus influenzae isolates (one resistant to ampicillin), five Streptococcus pneumoniae, five Neisseria meningitidis, one Klebsiella pneumoniae, one Pseudomonas aeruginosa, and one Listeria. In the chloramphenicol/ampicillin group, there were 19 H. influenzae isolates, 10 S. pneumoniae, three N. meningitidis, one Haemophilus parainfluenzae, and one Citrobacter. Of 63 patients with assessable CSF pathogens, one (3%) of 29 treated with sulbactam/ampicillin died (S. pneumoniae) and six (18%) of 34 treated with chloramphenicol/ampicillin died (two, H. influenzae; three, S. pneumoniae; and one, Citrobacter). Twelve percent in the sulbactam/ampicillin group and 18% in the chloramphenicol/ampicillin group had neurologic sequelae. No clinically significant reactions or toxicities were noted. Sulbactam/ampicillin was as effective as chloramphenicol/ampicillin in the treatment of meningitis. PMID- 3026016 TI - Clinical efficacy of sulbactam/ampicillin in pediatric infections caused by ampicillin-resistant or penicillin-resistant organisms. AB - Twenty-three children two months to 11 years old were treated with sulbactam/ampicillin or sulbactam/penicillin. Eleven had urinary tract infections (UTI), eight had pus-forming cervical adenitis, and four had lobar pneumonia. Pathogens were isolated from 18 patients: Escherichia coli from 10, Staphylococcus aureus from seven, and Klebsiella pneumoniae from one. All isolates were resistant to ampicillin or penicillin alone. Sulbactam (50 mg/kg per day) plus ampicillin (1:2 or 1:3 ratio) or penicillin (1:1.2 or 1:1.8 ratio) was given by intravenous bolus injection at 6-hr intervals for four to 11 days (mean duration, nine days). All pathogens were eradicated during treatment. Two patients with UTI relapsed after completion of treatment; the isolates were resistant to the combination. Clinical response was rapid and consistent with bacteriologic findings. Twenty-two of 23 children had a favorable clinical response. No systemic or local adverse effects were recorded. One child had eosinophilia and another had neutropenia at the end of treatment. Four children had slight and transient increases in hepatic transaminases. These results indicate that sulbactam/ampicillin may prove safe and effective for the treatment of non-life-threatening pediatric infections. PMID- 3026017 TI - A randomized comparative study of sulbactam plus ampicillin vs. metronidazole plus cefotaxime in the management of acute appendicitis in children. AB - Sulbactam is a beta-lactamase inhibitor that, when combined with ampicillin, gives the latter antibiotic a broad spectrum of activity, making it suitable for use as a prophylactic agent in acute appendicitis. In a single-blind, randomized trial, the efficacy of sulbactam plus ampicillin was compared with that of metronidazole plus cefotaxime. Thirty-five children undergoing appendectomy received intravenous sulbactam and ampicillin, while 38 children received metronidazole and cefotaxime. Single doses were given unless the appendix was considered gangrenous or perforated, in which case the drugs were administered for 72 hr. There were three wound infections in the group given sulbactam and ampicillin and five in the group given metronidazole and cefotaxime. The combination of sulbactam and ampicillin was well tolerated and appeared to be at least as effective as that of metronidazole and cefotaxime in the prevention of sepsis following appendectomy. PMID- 3026019 TI - Sulbactam plus ampicillin: interim review of efficacy and safety for therapeutic and prophylactic use. AB - The efficacy and safety of sulbactam/ampicillin has been evaluated in 39 studies of therapeutic use and six studies of prophylaxis. Studies of therapy were conducted in 899 patients: 751 seriously ill, many of whom had multiple concurrent diseases, and 148 with gonorrhea. Overall clinical and bacteriologic success was achieved in 92% of assessable cases; 88% of 768 pathogens in these patients were eradicated. Of these pathogens, 43% were resistant to ampicillin; eradication rates of 91% and 85% were achieved in ampicillin-resistant and ampicillin-sensitive organisms, respectively. In 388 patients who received prophylactic sulbactam/ampicillin, efficacy was similar to that of comparative agents and better than that of a placebo in preventing wound infections after appendiceal, biliary, upper-gastrointestinal, or gynecologic surgery. Adverse reactions were infrequent with the exception of injection-site pain, which occurred mainly after intramuscular injection and was reduced in incidence by concurrent administration of lidocaine. PMID- 3026018 TI - Efficacy and safety of sequential treatment with parenteral sulbactam/ampicillin and oral sultamicillin for skeletal infections in children. AB - Nine children with osteomyelitis and/or septic arthritis were treated sequentially with parenteral sulbactam/ampicillin and oral sultamicillin. Causative pathogens were identified in six cases; all were susceptible to the combination of ampicillin and sulbactam. The mean duration of parenteral therapy was 7.1 days (6-11 days), and the average hospital stay was 10.3 days (6-18 days). Peak serum bactericidal titers of greater than or equal to 1:8 were achieved in all patients during parenteral therapy; only one child receiving oral therapy did not achieve a titer of greater than or equal to 1:4. At follow-up, all of the children were cured clinically and there was no evidence of relapse. Adverse reactions to oral therapy were minimal. The regimen of parenteral sulbactam/ampicillin and oral sultamicillin used sequentially is effective and safe for the treatment of skeletal infections in children. The use of this approach significantly reduced the duration of hospitalization. PMID- 3026020 TI - [Tumors producer of hormones in the digestive system. Vasoactive intestinal polypeptide (VIP). Carcinoid]. PMID- 3026022 TI - Tumor markers and their clinical value. PMID- 3026023 TI - What to tell your patients about dietary fiber. PMID- 3026021 TI - Acute myocardial infarction in chronic Chagas' cardiomyopathy. Report of two cases with no obstructive coronary artery lesions. PMID- 3026024 TI - [Chin neuropathy in myeloma. Apropos of a new case]. PMID- 3026025 TI - [Neural tumors of the tongue. An anatomicoclinical study apropos of 13 cases]. AB - During the ten last years, 13 neural tumors of the tongue have been observed in the department of stomatology and maxillo-facial surgery of Salpetriere Hospital Paris (Prof. J.M. Vaillant): 4 amputation neuromas, 5 benign schwannomas, 1 malignant schwannoma, 2 neurofibromas and 1 multiple endocrine syndrome type III. The authors analyze the clinical and histological data from these case-reports; they point out difficulties encountered in the diagnosis, and the prognostic problems brought up by such infrequent lesions. PMID- 3026026 TI - Studies on the relationship between vitamin D3 status and urinary excretion of calcium in healthy subjects: effects of increased levels of 25-hydroxyvitamin D3. AB - The metabolic consequences of a rapid increase in vitamin D status in healthy subjects were investigated. Circulating levels of 25-hydroxyvitamin D3 were increased by 224% in 12 healthy men by giving oral vitamin D3 for 7 weeks and by 200% in 15 healthy women by UVB irradiation for 7 weeks. No statistically significant effects on the serum levels of calcium, phosphate, creatinine, urate, albumin, PTH, basal urinary excretion of calcium, fasting urinary excretion of cAMP, or urinary excretion of calcium after calcium load tests were observed with the unpaired t-test. With the paired t-test the small stimulatory effects (about 25%) on basal urinary excretion of calcium became statistically significant in both experiments. The ratio between calcium and creatinine in fasting urine was significantly elevated following UVB irradiation (from 0.11 +/- 0.02 to 0.21 +/- 0.04, p less than 0.025 unpaired t-test, p less than 0.02 paired t-test) but not after oral intake of vitamin D3. The level of 1,25-dihydroxyvitamin D in serum was not affected to a statistically significant degree by oral vitamin D3, whereas there was a slight decrease from 48 +/- 3 to 39 +/- 3 pmol/l following UVB irradiation. It is concluded that an increase in the concentration of 25 hydroxyvitamin D3 up to about 125 nmol/l has small and negligible effects on calcium homeostasis in healthy subjects. This finding is discussed in relation to our previous finding that hypercalciuric renal stone formers have elevated serum levels of 25-hydroxyvitamin D3 as compared with normocalciuric stone formers and healthy subjects. PMID- 3026027 TI - 25-Hydroxylase activity in subcellular fractions from human liver. Evidence for different rates of mitochondrial hydroxylation of vitamin D2 and D3. AB - 25-Hydroxylation of vitamin D2 and D3 was studied in subcellular fractions from human liver, using a technique based on isotope dilution-mass spectrometry. The mitochondrial fraction fortified with isocitrate catalysed 25-hydroxylation of vitamin D3 at a rate of about 10 pmol/mg protein X min. Under the same conditions, the rate of 25-hydroxylation of vitamin D2 was less than 2 pmol/mg protein X min. Crude microsomes fortified with NADPH catalysed 25-hydroxylation of vitamin D3 to a very low extent, and this activity was not linear with the amount of microsomal protein. A higher rate of conversion was obtained with a partially purified cytochrome P-450 fraction in the presence of NADPH-cytochrome P-450 reductase and NADPH. This fraction also catalysed 25-hydroxylation of 1 alpha-hydroxyvitamin D3 and 5 beta-cholestane-3 alpha, 7 alpha, 12 alpha-triol. 25-Hydroxylation of vitamin D2 could not be detected, neither with crude microsomes, nor with the microsomal cytochrome P-450 fraction. Since the assay for 25-hydroxyvitamin D2 was less sensitive than that for 25-hydroxyvitamin D3, these experiments do not rule out the presence of some 25-hydroxylase activity towards vitamin D2 in the microsomes. The results are discussed in relation to previous work in which a lower toxicity has been reported for vitamin D2 than for vitamin D3 in some mammalian species. PMID- 3026028 TI - Local immunoreactivity and human papillomavirus (HPV) in oral precancer and cancer lesions. AB - A retrospective longitudinal study was performed on 20 patients with oral leukoplakia, 10 of which developed an oral squamous cell carcinoma, to assess whether any alterations in the local immunologic reactivity could be found of value in predicting the subsequent behavior of the lesions. During the major period of follow-up, the relative frequency of in situ IgA-producing plasma cells was significantly higher in biopsies from patients subsequently developing cancer than in patients showing no cancer development. Preceding the malignant transformation by 12 to 15 months, however, a remarkable shift from IgA to IgG plasma cell predominance was noticed in the biopsies of the cancer series, not detectable in the non-cancer group. HPV group specific capsid antigens were found in seven cases of the cancer series and in six of the non-cancer group. The possible diagnostic implications of these results are discussed. PMID- 3026029 TI - The study of biominerals by high resolution transmission electron microscopy. AB - This paper presents an overview of the study of the ultrastructure of biogenic inorganic solids (biominerals) using high resolution transmission electron microscopy (HRTEM). A range of biominerals have been studied including iron oxides, calcium phosphates, calcium carbonates and silica. The studies have revealed information concerning the structural complexity of these materials and have identified crystallographic order and disorder at the nanometre level. In addition, the results have aided the elucidation of the mechanisms of nucleation and growth of biogenic minerals. PMID- 3026030 TI - The use of a line scan ratemeter for the X-ray microanalytic evaluation of membrane-bound histochemical endproducts. AB - Although X-ray microanalysis represents a useful tool for identifying electron dense histochemical end-products, quantitative microanalytic measurements are seriously hampered in the case of the activities of certain membrane-bound enzymes. For example, the electron histochemical methods revealing K+-dependent pNPPase activity result in a very fine, granular reaction product of lead phosphate. Therefore microanalytic, densitometric of similar evaluations of the reaction, even in semiquantitative terms are not practical by the usual procedures. This paper describes a method of X-ray microanalysis of thick sections (0.5 micron) processed for K+-pNPPase, where a sufficient amount of lead is present for X-ray microanalytic determination. The analysis is performed in the line scan mode on transversely cut membrane profiles by means of the line scan ratemeter of an EDAX System F. This yields quantitative data on the relative lead concentrations in the vicinity of the cell membrane. A method is proposed for calculation of relative enzyme activities based on the Pb-signal of the ratemeter curve and the average "noise"-level of the cytoplasm, containing also non-specifically bound lead. This method avoids the necessity of measuring the section thickness; it may be useful for a variety of purposes in the electron microscopic histochemistry of membrane-bound enzymes. PMID- 3026031 TI - Urolithiasis in a patient ingesting pure silica: a scanning electron microscopy study. AB - A patient who repeatedly produced urinary calculi, had consumed about 3 g of cristobalite (SiO2) per day for many years. Investigations using scanning electron microscopy revealed minute particles containing silicon in the core of the stone as well as in urine sediment. A mechanism similar to that proposed for the effect of silicon-containing drugs against gastric ulcer, may play a role in this formation of silicon-containing urinary stones. PMID- 3026032 TI - Urinary calculi and urinary tract infection. A clinical and microbiological study. AB - The problem of urinary calculi in association with urinary tract infection (UTI) was investigated. Fifty-two (7%) of 796 patients attending our outpatient stone clinic had UTI which was considered of pathogenic importance for their stone formation. Proteus was the most common microorganism. Metabolic disorders were found in one third and anatomical and functional abnormalities in two thirds of the patients. The infected patients had lower urinary calcium excretion and higher serum creatinine than idiopathic stone formers and had a higher frequency of stone operations. The prevalence of staghorn calculi as a cause of urimia was low (1.5% of 481 dialysis patients). Eight such patients were studied and six of them had metabolic and anatomical disorders. The time taken for the uremia to develop was 7.4 +/- 2.0 (SD) years. In 535 patients treated surgically for renal calculi, about one third had positive urine culture at the time of operation and E. coli was the most common bacterial strain (35%). Proteus was found in 28% and these patients had the highest frequency of UTI episodes, most of which occurred before hospitalization. Patients infected with E. coli had a higher frequency of phosphate-containing calculi than non-infected patients, in whom the highest frequency of calcium oxalate calculi was found. A new broad-spectrum cephalosporin, ceftazidime, was used as perioperative prophylaxis in 15 patients operated upon for renal calculi and UTI. Ten had bacterial growth in the renal pelvis and all strains were eradicated. Bacterial growth was found in two out of six cultured stones from patients with bacterial growth in the pelvis. The pharmacokinetics of the drug was studied and the decreases in the ceftazidime levels in serum and renal tissue seemed to be parallel. Bacterial binding of urinary isolates to hydroxyapatite (HAP) particles was studied. Two E. coli strains (A5089 and E7704) and one Proteus mirabilis strain (A5076), all obtained from stone patients, were compared with two E. coli strains (2683 and M7810) with well defined cell-surface properties. Hemagglutination tests were performed and the cell-surface hydrophobicity was determined by a salt-aggregating test. When the strains were cultured at 37 degrees C to promote fimbriae formation they hemagglutinated erythrocytes and displayed hydrophobic cell-surface properties, and showed higher capacity for binding to HAP than when cultured at 18 degrees C to suppress fimbriae formation, when they showed almost complete absence of hemagglutination and low cell-surface hydrophobicity. Bacterial cell-surface properties seem to influence the binding of uropathogens to HAP particles. PMID- 3026033 TI - Leukotriene B4, a mediator of inflammation? PMID- 3026034 TI - Increased collagenase activity in human rheumatoid meniscus. AB - Collagenase activity of the knee joint menisci of patients suffering from rheumatoid arthritis was approximately 3-fold higher than that found in menisci of control patients. The mean collagenase activity in the macroscopically more diseased parts of the rheumatoid menisci was significantly higher than that in the less damaged areas. The specific degradation products resulting from the cleavage of human meniscoid type II collagen by rheumatoid meniscoid collagenase were demonstrated by SDS-polyacrylamide gel electrophoresis. Addition of N ethylmaleimide, which activates latent mammalian collagenases, did not further increase collagenase activity in rheumatoid menisci. Thus in rheumatoid meniscus, collagenase may be synthesized and then activated, probably by proteolytic enzymes involved in the inflammatory reaction. PMID- 3026035 TI - Environmental surveys in the European man-made mineral fiber production industry. AB - This paper presents estimates of airborne fiber concentrations and fiber size for European man-made mineral fiber (MMMF) factories on the basis of measurements made in 1977-1980. The airborne fiber concentrations previously reported at a conference of the World Health Organization (WHO) and the International Agency for Research on Cancer in 1982 have been revised to harmonize the results with the WHO-European MMMF reference counting level. The result was an approximate doubling of the reported airborne fiber levels. After the revisions the average combined occupational group concentrations in the rock- and glass-wool plants were still generally low (less than 0.01 fibers/ml). In the glass continuous filament factories the airborne fiber concentrations were very low (less than 0.01 fibers/ml). The average plant median for fiber length ranged from 10 to 20 microns, and the corresponding median diameters ranged from 0.7 to 2 microns. In general the glass-wool fibers were thinner than the rock-wool fibers. The fiber concentrations measured in other studies in the MMMF production and user industries are reviewed. Higher levels (between 0.1 and 1.0 fibers/ml) have been measured in some insulation wool production, secondary production, and user industries. The highest levels (greater than 1.0 fibers/ml) occurred in very fine glass-fiber production and in other specialist insulation wool usage. PMID- 3026036 TI - Past exposures to airborne fibers and other potential risk factors in the European man-made mineral fiber production industry. AB - A historical environmental investigation was undertaken in European man-made mineral fiber factories (MMMF). The aim was to assess past exposures to MMMF and other environmental risk factors (asbestos, polycyclic aromatic hydrocarbons, polychlorinated biphenyls, formaldehyde, and arsenic). A self-administered questionnaire completed by each plant management and an interview of the respondents were used. Addition of oil to the MMMF, change in the nominal fiber size of the bulk MMMF, and elimination of early discontinuous production techniques were identified as principal changes. The absence of oil, small nominal size, and labor-intensive production have been judged to be associated with higher airborne fiber levels. It was concluded that for epidemiologic purposes the subdivision of the history of each plant into an early, intermediate, and late technological phase based on the aforementioned changes would be appropriate. Corresponding time periods were identified for asbestos usage, use of bitumen as a binder, and production of slag wool. Two factories were singled out for specific attention: one where asbestos was used in calcium silicate brick production and one where olivine, used as a raw material, may have been contaminated with natural mineral fibers and where the production process would have exposed workers to coal-tar pitch volatiles. PMID- 3026037 TI - Contributions to the IARC (International Agency for Research on Cancer) study on mortality and cancer incidence among man-made mineral fiber production workers. PMID- 3026038 TI - Incidence of cancer in the mineral-wool producing industry in Norway. AB - This study concerned the Norwegian phase of a European collaborative investigation on workers in man-made mineral fiber production. A study population of 2,361 men from four Norwegian plants was examined for mortality and cancer incidence, especially lung cancer, based on a comparison of observed and expected figures, the latter determined according to the five-year age-specific mortality and incidence rates for the entire country. Violent deaths among workers with less than one year of employment represented the only significant mortality excess. A borderline, statistically significant excess risk was found for cancer of the buccal cavity and pharynx, but the fact that the excess occurred in one factory only led to the assumption that the risk was associated with factors other than mineral wool. An excess risk for lung cancer was also found among those with 20 years or more since first exposure (9 observed and 4.36 expected). The risk that emerged was presumably initiated before 1960, when the environmental conditions were more hazardous than later. PMID- 3026039 TI - [Intestinal absorption of calcium gluconate and oseine-mineral complex: an evaluation by conventional analyses]. AB - Calcium (Ca) preparations are widely used in the treatment of osteoporosis, usually as soluble salts. Tolerance might be improved by prescription of slowly dissolved Ca preparations, since Ca is also absorbed distally, even in the colon. In this regard the use of natural forms of Ca might be advantageous, but natural products cannot be labeled reliably for easy evaluation of their absorption. To study the intestinal absorption of an osseino-mineral complex (Ossopan) in comparison with Ca-gluconate, healthy males were investigated by means of conventional blood and urinary measurements before and after ingestion of either substance containing 1.58 g Ca. All subjects were placed on a standard diet 2 days before and during test day. Ca-gluconate (n = 7) evoked a marked and transient rise in plasma ionized calcium; total plasma calcium and urinary calcium followed a parallel course, while plasma and urinary phosphate decreased. After administration of osseino-mineral complex (n = 6), a slow but sustained elevation of plasma ionized calcium was observed while total calcium remained unchanged when corrected by the plasma proteins. Plasma phosphate and proteins increased, as did urinary phosphate. Comparing the 24-hour urine of the test day with that of the previous day, the rise in calcium excretion was slightly greater in the subjects treated by osseino-mineral complex than in those who were given Ca-gluconate, while phosphate excretion increased in the first group and decreased in the second. It is concluded that the bioavailability of Ca in osseino-mineral complex is as good as, if not better than, that of Ca gluconate.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3026040 TI - [Are non-bovine Artiodactyla IBR virus reservoirs? I. BHV-1 and CapHV-1 infection and reactivation studies to show virus type specificity of humoral antibodies and characterization of viral antigens]. PMID- 3026041 TI - [Are non-bovine Artiodactyla IBR virus reservoirs? II. Seroepidemiologic studies on goats, sheep, swine and wild Artiodactyla in Switzerland]. PMID- 3026042 TI - [Recent advances in the study of atrial natriuretic factor]. PMID- 3026043 TI - [Recent advances in adenylate cyclase research]. PMID- 3026044 TI - [Endogenous digitalis-like substances]. PMID- 3026045 TI - [Molecular biology of hemopoietic regulators]. PMID- 3026046 TI - [Alpha-receptors in the heart and ischemic arrhythmias]. PMID- 3026047 TI - [Factors influencing the secretion of pulmonary surface active substances]. PMID- 3026048 TI - The atomic structure of Mengo virus at 3.0 A resolution. AB - The structure of Mengo virus, a representative member of the cardio picornaviruses, is substantially different from the structures of rhino- and polioviruses. The structure of Mengo virus was solved with the use of human rhinovirus 14 as an 8 A resolution structural approximation. Phase information was then extended to 3 A resolution by use of the icosahedral symmetry. This procedure gives promise that many other virus structures also can be determined without the use of the isomorphous replacement technique. Although the organization of the major capsid proteins VP1, VP2, and VP3 of Mengo virus is essentially the same as in rhino- and polioviruses, large insertions and deletions, mostly in VP1, radically alter the surface features. In particular, the putative receptor binding "canyon" of human rhinovirus 14 becomes a deep "pit" in Mengo virus because of polypeptide insertions in VP1 that fill part of the canyon. The minor capsid peptide, VP4, is completely internal in Mengo virus, but its association with the other capsid proteins is substantially different from that in rhino- or poliovirus. However, its carboxyl terminus is located at a position similar to that in human rhinovirus 14 and poliovirus, suggesting the same autocatalytic cleavage of VP0 to VP4 and VP2 takes place during assembly in all these picornaviruses. PMID- 3026049 TI - Myosin rod phosphorylation and the catch state of molluscan muscles. AB - "Catch" is a prolonged state of tension in molluscan smooth muscles shown by mechanical measurements to be associated with the level of protein phosphorylation. Myosin isolated from these muscles is unusual in being phosphorylated in the rod portion by an endogenous kinase, like certain nonmuscle myosins. These findings suggest that the myosin rod is a target for phosphorylation and that this reaction may control the transition from catch to relaxation. PMID- 3026050 TI - The lung as a metabolic organ. AB - Recently, the lung has received increasing attention as a metabolic organ. In this role, the lung modulates the composition of the arterial blood by several mechanisms: removing active substances from the plasma, releasing substances into the plasma, temporarily holding substances from circulation, and activating or inactivating substances that pass through the lungs. In this report, the procedures proposed by different investigators for in vivo noninvasive assessment of the lung metabolic functions are reviewed. Most procedures are based on an estimation of the clearance of plasma amines by the lung endothelial cells. This clearance is assessed by measuring the lung uptake or the extraction fraction of an intravenously (IV) injected radiolabeled amine. Our own procedure, which assesses the number of free pulmonary endothelial amine receptors, is discussed in detail. In our procedure, the number of receptors was computed using the number of injected molecules of amine and determining the lung extraction fraction of the amine during its first pass through the lungs. In goats, using N isopropyl-p-iodoamphetamine labeled with 123I as the radiopharmaceutical, the total number of endothelial lung amine receptors was found to be 1.589 X 10(20). The methods for studying the lung metabolic functions, which are discussed in this report can be applied in humans to evaluate either physiological or pathological conditions. PMID- 3026051 TI - Progestational agents in advanced breast cancer: an overview. AB - Progestational agents, such as megestrol acetate and medroxyprogesterone acetate, are effective hormonal treatments for metastatic breast cancer in postmenopausal women. Clinical trials of these agents have demonstrated that 30% to 60% of patients will experience objective tumor response, depending on pretreatment prognostic variables. Although optimal doses and schedules have not been fully defined, current studies are investigating the therapeutic effectiveness of high dose progestins. Toxicity of these drugs is mild and generally limited to weight gain related to their anabolic activity. Progestins are active second-line agents for metastatic breast cancer in postmenopausal women. In selected patients, they appear to be equivalent to tamoxifen as first-line therapy for metastatic disease. As more patients are exposed to prolonged adjuvant tamoxifen therapy, the role of progestins as first-line hormonal therapy at the time of relapse is likely to expand. PMID- 3026052 TI - Megestrol acetate v tamoxifen in advanced breast cancer: correlation of hormone receptors and response. AB - This report describes the preliminary results from a randomized study comparing megestrol acetate with tamoxifen in the treatment of postmenopausal women with advanced breast cancer, correlating estrogen receptors (ER) and progesterone receptor (PgR) status with response. Patients received megestrol acetate (40 mg) orally four times daily or tamoxifen (10 mg) orally twice daily. If the initial therapy failed, patients were crossed over to the alternate treatment. Of 197 patients entered in the study, 190 were considered evaluable. The overall response rates were 35% with megestrol acetate and 42% with tamoxifen. Twenty three percent (7/30) of patients responded to megestrol acetate after being crossed over from tamoxifen, while 22% (6/27) responded to tamoxifen after being crossed over from megestrol acetate. Response did not correlate significantly with combined receptor (ie, ER plus PgR) levels. A significant trend was seen between ER level and response only in the tamoxifen group. There was no association between PgR levels and response for either tamoxifen or megestrol acetate. PMID- 3026053 TI - [Analgesic effect of septal stimulation and the intraseptal opiate receptor on visceral pain]. PMID- 3026054 TI - [Effect of PGE2 on the cyclic nucleotide content of isolated neonatal rat pancreatic islets treated with alloxan]. PMID- 3026055 TI - [Effect of tyrosine on testosterone and cAMP production by Leydig cells in vitro]. PMID- 3026057 TI - Prognostic factors in patients with small-cell carcinoma of the lung. AB - Fifty-two patients with histologically confirmed small-cell carcinoma of the lung, 15 with disease localized to one hemithorax and 37 with extensive metastases, were studied. Only patients whose general condition was considered good enough to tolerate intensive combination chemotherapy were included in this study. The median survival time of these patients was 33 weeks (range 7-192 weeks) with a mean survival time of 43 weeks. Patients who responded with objective tumour improvement within 6 weeks had a median survival time of 39 weeks. Those showing disease stabilization at 6 weeks had a median survival time of 35 weeks while those having disease progression at this time had a median survival of 22 weeks. Patients without liver involvement and a normal serum albumin level at the start of treatment had statistically superior survival times. PMID- 3026058 TI - Carcinoma related to choledochal cysts with internal drainage operations. AB - Carcinoma arising in a choledochal cyst retained after enteric drainage is discussed in comparison with primary carcinoma. In our series, carcinoma developed in eight of 82 patients with choledochal cyst and three of the eight had secondary carcinoma occurring several years after enteric drainage. Forty-two similar instances and 235 instances of primary carcinoma have been reported in Japan. The mean age at detection of carcinoma in enteric drainage was 35 years old, 15 years less than that of primary carcinoma. The mean interval between drainage and detection of carcinoma was ten years. Carcinoma mainly arose from the retained cyst in drainage, while primary carcinoma developed in either the cyst or gallbladder. The prognosis was very poor. Almost all patients undergoing enteric drainage died soon after the detection of carcinoma. This is possibly due to the delay in diagnosis. Pancreatic juice can easily regurgitate into the choledochus through an anomalous junction of the pancreatobiliary ductal system, and enteric drainage causes pancreatic juice in the cyst to become activated due to the influx of intestinal juice. Therefore, inflammatory changes of the cyst are accelerated and probably result in carcinoma. Early excision of the choledocus should be recommended in patients with choledochal cyst retained after enteric drainage. PMID- 3026059 TI - Four new hepatectomy procedures for resection of the right hepatic vein and preservation of the inferior right hepatic vein. AB - Among the accessory hepatic veins, the thickest one is the IRHV and is a significant vessel in 20 to 24 per cent of the patients. In these patients, if the right hepatic vein is totally resected, the right posteroinferior area can be preserved with the IRHV. Four types of hepatectomies n which the IRHV was preserved were proposed and three of the four procedures were performed upon five patients. In these five operations, the hepatectomies were successfully performed and blood losses were from 1,020 to 3,200 milliliters. These operative procedures, which have not been described before, could widen the indication of hepatectomy in patients with reduced liver function and tumor involving the right hepatic vein. In order to perform these operative procedures successfully, intraoperative sonography is indispensable. PMID- 3026060 TI - Colectomy with ileorectal anastomosis for familial adenomatous polyposis: the risk of rectal cancer. AB - One hundred thirty-three patients with familial adenomatous polyposis undergoing colectomy and ileorectal anastomosis have been reviewed for the occurrence of cancer in the rectal stump. Ten patients developed rectal cancer (Actuarial survivorship rate of 88% for those patients free of rectal cancer at 20 years). Potential risk factors for the development of rectal cancer, including age at colectomy, previous colon cancer, number of rectal polyps, and length of the rectal stump, were analyzed and no significant differences were found. A policy of total colectomy with ileorectal anastomosis at 12 to 15 cm with conscientious lifelong follow-up thereafter is advocated for persons affected by familial adenomatous polyposis. PMID- 3026062 TI - [Functional activity of neutrophils in patients with abdominal typhus]. AB - Cytochemistry and cytofluorometry were employed to study the main components of the intraleukocyte microbicide system of neutrophils in 52 patients with typhoid fever. During the disease there was a consistent decrease in the content of cationic protein and in the activity of leukocyte myeloperoxidase depending on the stage and severity of the pathological process. At the same time an increase in the glycogen content and activation of acid and alkaline phosphatases of leukocytes with a maximum at the disease height were detected. Addition of the complications aggravated cytochemical alterations in the cell and led to the delay in their elimination. Profound qualitative and quantitative changes in the activity of intracellular components with a visible clinical recovery of the patients pointed to an unfavourable prognosis. PMID- 3026061 TI - [Status of the hypophysis-adrenal cortex system during treatment with glucocorticoids by the method of intermittent (every other day) administration]. AB - The authors discuss the advantages of the use of glucocorticoids in the treatment of autoimmune diseases, if the drugs are administered every other day. A total of 149 patients with autoimmune thyroiditis, endocrine ophthalmopathy, myasthenia and bronchial asthma were examined. Of these, 3 patients received glucocorticoids every other day, whereas the remainder took them every day. The former method consisted in the use of a single daily dose of glucocorticoids (20 to 80 mg) at 7 to 8 o'clock in the morning over 2 months to 5 years. The function of the pituitary-adrenal system was tested by the measurement of the blood content of ACTH, hydrocortisone and testosterone with the aid of a kit of standard tests. It was shown that glucocorticoids administered every other day did not produce any suppression of the pituitary-adrenal system in contrast to those who received prednisolone every day. The lack of the side effects in the form of exogenous hypercorticoidism can be explained by the anabolic action of steroid hormones produced by the reticular zone of the adrenal cortex. Examination of the blood serum content of testosterone in women who received glucocorticoids every other day has demonstrated that modulations in its secretion were similar to those in the content of ACTH and hydrocortisone. Thus, the stimulating hormonal therapy with glucocorticoids administered every other day appears more physiological as compared to the daily schedule. PMID- 3026063 TI - [Hypercalcemia in childhood. A boy with asymptomatic hypocalciuric hypercalcemia]. AB - The diagnostic work-up of a patient with (familial) hypocalciuric hypercalcemia (FHH) is discussed. The patient showed no clinical signs of hypercalcemia. There were no indications of vitamin D intoxication. In the first degree family members no hypercalcemia was found. Physical examination was normal. In contrast to hyperparathyroidism, FHH is usually symptomless. Furthermore, FHH is characterized by a normal chloride/phosphate ratio, a normal, but relatively high, serum parathyroid hormone level, a relatively low urinary calcium excretion, a calcium-creatinine clearance ratio less than 0.01 and a normal cyclic AMP excretion. PMID- 3026064 TI - The mode of reduction of vanadate(+V) to oxovanadium(+IV) by glutathione and cysteine. AB - Vanadate(+V) is completely reduced to oxovanadium(+IV) in vitro by thiols. A high transient absorbance at 750 nm is observed (intense blue) in solutions of a pH below 5 immediately after starting the reduction. EPR-measurements and determinations of the consumption of free thiol groups prove that the transient absorbance is caused by an intermediate compound not containing vanadium in its reduced state (+IV). The way of reduction of vanadate(+V) by thiols is discussed. We propose the formation of an intermediate vanadate(+V)-thioester which is formed before reduction in a preequilibrium step. PMID- 3026065 TI - New evidence on the nature of hypocorticism in thymomegaly. AB - According to the present clinical-experimental investigations, hypocorticism at the thymomegaly (hyperplasia of the thymus) was shown to occur as the secondary phenomenon as a result of the reduction of the corticotrophin function of adenohypophysis due to the activation of dopaminergic and serotoninergic central inhibition systems. PMID- 3026066 TI - The effects of uremic serum and 3'-5' cyclic AMP on blastogenesis of normal lymphocytes. AB - Phytohemagglutinin (PHA) induced lymphocyte blast transformation is impaired both in uremic lymphocytes and in normal lymphocytes exposed to uremic serum. Cyclic AMP is known to inhibit blast transformation in normal and uremic lymphocytes. This investigation was undertaken to assess quantitatively the effects of uremic serum and cyclic AMP on blastogenesis of normal lymphocytes. Uremic serum or cyclic AMP significantly inhibited blast transformation of normal lymphocytes. These effects were statistically similar and cumulative. We conclude that the inhibition imposed by uremic serum on normal lymphocyte blastogenesis is predominantly mediated by a mechanism(s) different from cyclic AMP. PMID- 3026067 TI - Aqueous production. AB - The formation of aqueous humour by the ciliary body is a complex process. Active transport of solutes by the ciliary process epithelium is an energy-dependent mechanism that selectively transports substances against an electrochemical gradient across the cell membranes. Water passively follows the active solute transport. In addition to these active transport processes, ultrafiltration contributes to the formation of aqueous humour. The ciliary epithelium contains enzyme systems that function in the production of aqueous humour. The enzymes sodium-potassium-activated adenosine triphosphatase [(Na+:K+)ATPase] and carbonic anhydrase participate in the active transport across this epithelium. Inhibition of these enzymes lowers intraocular pressure (IOP) by decreasing aqueous humour production. the ciliary epithelium contains both alpha- and beta-adrenergic receptors. Electrophysiologic studies on the isolated iris-ciliary body (I-CB) preparation provide a means to study direct effects of the adrenergic agents on transepithelial properties of the ciliary epithelium. This paper will discuss the enzymatic and adrenergic properties of the ciliary epithelium as they relate to active transport and thereby aqueous humour production. PMID- 3026069 TI - Disappearance of Chikungunya virus from India and South East Asia. PMID- 3026068 TI - The ophthalmological features of AIDS and AIDS related disorders. AB - The ocular findings in 38 patients infected by the Human T Cell Lymphotropic Virus Type III (HTLV III) are reviewed. Twenty patients were suffering from the Acquired Immunodeficiency Syndrome (AIDS) and 18 from the HTLV III related disease Persistent Generalised Lymphadenopathy (PGL). Cotton wool spots were found in the retinae of 8 patients at some stage of their disease and the prognostic significance of these is discussed. Cytomegalovirus (CMV) Retinitis was an ocular complication in eight of the patients with AIDS. The therapeutic effects of a new anti-viral drug, di-hydroxy propoxymethyl guanine (DHPG), are discussed. PMID- 3026070 TI - Late-onset hemorrhagic cystitis associated with urinary excretion of polyomaviruses after bone marrow transplantation. AB - Hemorrhagic cystitis is a well known complication of allogeneic bone marrow transplantation (BMT) and is normally attributed to the use of high-dose cyclophosphamide in the preparative regimen. Hemorrhagic cystitis occurring late after BMT is unlikely to be due to the effects of this conditioning, and probably has an infective etiology. Three patients undergoing BMT for chronic granulocytic leukemia (CGL) developed terminal dysuria and hematuria at 38, 56, and 149 days post-BMT. Electron microscopy (EM) of urine voided at these times revealed large numbers of papovavirions, which were subsequently identified as BK virus. Urine samples inoculated onto human embryonic lung fibroblasts induced infection of the cells and replication of the virus as detected by EM of tissue culture fluid. Urine from one of these patients was examined by standard cytological techniques, and EM of urothelial cells showed nuclear inclusions consisting of nonencapsulated virus particles of diameter 40 nm, consistent with papovavirus. Five further patients were found to be excreting BK virus without symptoms of cystitis, although one of these patients did experience abnormalities of liver function that were otherwise unexplained. BK virus has already been implicated in hepatic dysfunction posttransplant, and in cystitis in nonimmunosuppressed children. We postulate that it may also be involved in the etiology of late hemorrhagic cystitis after BMT. PMID- 3026071 TI - Loss of myocardial viability following hypothermic perfusion storage from contaminating trace elements in the perfusate. AB - Two groups (A and B) of isolated baboon hearts were preserved by continuous hypothermic perfusion storage for 48 hours using perfusates that, according to the manufacturers, differed only in the concentrations of the contaminating trace elements iron, lead, and arsenic. Storage with the perfusate containing the higher concentration of these elements (perfusate B) led to significantly less gain in heart mass, a greater reduction in coronary flow, coronary sinus effluent lactate, and myocardial arteriovenous oxygen difference and a greater increase in coronary sinus effluent lactate dehydrogenase, when compared with perfusate A. Group B hearts totally failed to support the circulation following orthotopic transplantation, whereas group A hearts showed excellent function. Group B hearts had undergone the typical changes of enhanced resting myocardial tension during the storage period (before warm blood reperfusion); we proposed that these changes were brought about by the production of superoxide anions and radicals by the higher relative concentration of iron, or a combination of contaminating trace elements, in perfusate B. To confirm that these perfusates did differ significantly in the concentration of these trace elements, in particular with regard to iron, the superoxide anion activity in both solutions was measured and was found to be significantly higher in perfusate B. The addition of superoxide dismutase to both solutions inhibited superoxide anion activity by more than 80%. PMID- 3026072 TI - Percutaneous interstitial chemotherapy of a small hepatocellular carcinoma under ultrasound guidance. AB - A small (less than 3 cm) inoperable hepatocellular carcinoma was treated with percutaneous interstitial chemotherapy (PIC). 5-Fluorouracil was injected by a fine needle under ultrasound guidance. After 3 months a fine needle biopsy (FNB) yielded fibronecrotic material. After 18 months another FNB yielded steatosis and dysplastic cells and the lesion showed no increase in size. PIC could be an interesting alternative treatment for small tumors unresponsive to conventional therapies. PMID- 3026073 TI - [Effect of proteolytic enzymes on cyclic nucleotide levels and DNA synthesis in lymphocytes]. AB - It is shown that the treatment of mouse spleen lymphocytes with the optimum mitogenic concentrations of thrombin and plasmin increases the intracellular amount of cGMP, the intracellular level of cAMP remaining the same. An increase of the intracellular level of cGMP and enhancement of the DNA synthesis in lymphocytes, on the one hand and the absence of these metabolic changes with the use of inactivated proteases, on the other, indicate that thrombin- and plasmin induced proliferation of lymphocytes at the initial stages of cell activation is, probably, realized by the cGMP-dependent mechanisms. PMID- 3026074 TI - [Endogenous ouabain-like substances in animal tissues]. AB - Methods of purification and properties of the digitalis-like substances from different animal-origin sources are considered. Possible mechanisms of the cardiac glycoside-effect on the cardiac muscle and endogenous digitalis-like factors' effect on the smooth muscles of the blood vessels are discussed. Criteria that may be used to characterize the digitalis-like activity in samples and extracts obtained from different tissues are presented. The role of these substances in some types of pathologies, especially in the development of hypertension, is considered. PMID- 3026076 TI - Prostatic invasion of rectal carcinoma: does transurethral surgery alter prognosis? AB - Two cases of colorectal carcinoma invading the prostate are presented. Each patient initially had an abdominoperineal resection for colorectal carcinoma followed by a transurethral resection of the prostate (TURP) for benign prostatic disease. In both patients colonic carcinoma developed subsequently in their prostatic fossae. Pathogenesis of late tumor invasion into the prostate is reviewed and consideration given to a more conservative surgical approach. PMID- 3026077 TI - [Surgical treatment of lung cancer in young patients]. AB - The morphological structure of tumors is shown to be the leading factor responsible for the volume of the operative treatment in young patients. The improvement of organization of mass prophylactic x-ray fluorography in young persons aimed at early diagnosis of lung carcinoma is a necessary condition of obtaining better surgical results. PMID- 3026078 TI - A bovine respiratory virus vaccination trial. AB - A respiratory virus vaccination trial was carried out in a commercial calf rearing unit with a history of virus pneumonia. The effects of vaccination on the incidence of virus respiratory disease and growth rate were assessed. Forty-four bought-in calves were allocated to groups and treated as follows: A, unvaccinated controls; B, intranasal temperature-sensitive infectious bovine rhinotracheitis (IBR) vaccine at three and 10 weeks; C, intranasal temperature-sensitive combined IBR and parainfluenza-3 (PI3) vaccine at three and 10 weeks; D, intranasal temperature-sensitive combined IBR and PI3 vaccine at three and 10 weeks plus live attenuated bovine respiratory syncytial (BRS) virus vaccine intramuscularly at seven, 10 and 16 weeks. Two outbreaks of virus pneumonia occurred, one at three to four months of age associated with BRS virus and the other at four to five months of age with PI3 virus. During these outbreaks the incidence of pneumonia was lower and the number of days of elevated temperature and the number of treatments were significantly less in groups vaccinated against the associated virus. Despite these findings there were no significant differences between the growth rates of the groups either during the outbreaks of virus pneumonia or during the 10 month period to slaughter. PMID- 3026079 TI - Control of feline leukaemia virus infection by a removal programme. PMID- 3026075 TI - Structure and function of the epididymis. AB - Testicular spermatozoa are functionally immature in that they cannot fertilize ova. It was first demonstrated by Young that spermatozoa undergo certain changes as they migrate through the epididymis. He proposed that spermatozoa ripen during epididymal transit. It is now known that specific maturational changes occur in spermatozoa during epididymal transit which result in their developing the ability to fertilize ova. Concomitant with this functional maturity are changes in spermatozoal morphology, motility, chemistry, permeability, density and metabolism. It is apparent that in some way not understood these changes are necessary for sperm to achieve the ability to complete the fertilization process. When these mechanisms are understood, we may be able to effectively treat conditions such as necrospermia or abnormally low sperm motility. Furthermore, with the development of the hamster-egg penetration test a "new" type of male infertility has become evident in recent years; the inability of otherwise normal sperm to penetrate an ovum. It is during epididymal transit that this ability is normally acquired. Thus, any insight into how sperm attain the capacity to penetrate an ovum could lead to an effective treatment of patients whose sperm do not have this ability. In addition, the epididymis holds significant promise as the site of action for a male contraceptive. Thus, it is the purpose of this review to describe the structure and function of the mammalian epididymis with particular emphasis on the factors regulating sperm maturation. PMID- 3026080 TI - Immunocompetence of sheep experimentally infected with bovine leukemia virus. AB - The humoral and cellular immunological reactivity of sheep were studied throughout the first 32 weeks following experimental infection with bovine leukemia virus (BLV). Seroconversion of BLV-inoculated sheep occurred within 4 weeks, but infection was not transmitted to contact control sheep. Despite the persistence of the viral infection, no differences were demonstrated in leukograms, serum IgG concentrations, humoral response to immunization with an irrelevant antigen (rabbit red blood cells), phytomitogen (Concanavalin A and Pokeweek mitogen)-induced lymphocyte blastogenesis, or chemical (1-chloro, 2-4 dinitrobenzene) skin contact hypersensitivity, between BLV-infected and uninfected contact control sheep. These results demonstrate the absence of a nonspecific immunosuppressive effect of BLV and further negate the influence of a generalized immunological deficit on the development of clinical disease in BLV infected animals. PMID- 3026081 TI - Cell-mediated cytotoxicity of peripheral blood mononuclear cells stimulated in vitro for infectious bovine rhinotracheitis virus-infected cells. AB - Peripheral blood mononuclear cells (PBMC) from calves infected with and hyperimmunized to infectious bovine rhinotracheitis virus (IBRV) were stimulated in vitro with viral antigens to evaluate their cytotoxicity for a variety of cells. The 51-Cr release assay was used to measure cytotoxicity. Cytotoxicity was not present in fresh nonstimulated cells, but was detected in cultured, IBRV stimulated cells at day 3, was maximal at day 7, and declined thereafter. PBMC stimulated in vitro with IBRV expressed a preference for killing IBRV-infected cells compared to pseudorabies virus (PRV)-infected cells. IBRV-infected, but not PRV-infected, cold target cells inhibited lysis of IBRV-labeled target cells. High concentrations of IBRV hyperimmune serum partially blocked cytotoxicity. Cells expressing a viral preference for cytotoxicity showed no preference for lysis of autologous compared to heterologous bovine cells. PBMC from calves that were either IBRV-immune or not immune were cultured without IBRV stimulation and had similar levels of cytotoxicity for IBRV-infected cells as cells from IBRV infected cattle. PMID- 3026082 TI - Different types of microcalcifications observed in breast pathology. Correlations with histopathological diagnosis and radiological examination of operative specimens. AB - Microcalcifications taken from 50 systematized mammary excisions were submitted to light microscopic and scanning electron microscope analysis. Microprobe and x ray diffraction analyses were also performed. Two main types were observed: Type I microcalcifications composed of weddellite crystals. They were observed in benign breast lesions only (11 cases out of 21) or, in lobular carcinomas in situ (L.C.I.S.) of the breast (5 cases out of 6). They were not seen in 3 cases of intraductal carcinoma (I.D.C.) nor in infiltrating (I.C.) carcinomas (20 cases). Type II microcalcifications, non-cristalline in nature, composed of calcium, phosphate, hydroxyapatite or of phosphorus and calcium associated with other elements, were observed in benign lesions (10 cases out of 21) and in all cases of infiltrating carcinomas. The microcalcifications observed on mammography were also found on the radiographs of systematised mammary excisions from the lesion or from its immediate vicinity, but only when using the appropriate technique. Microcalcifications are therefore an excellent marker of breast lesions but they cannot be simply divided into "benign" or "malignant" types. Nevertheless, the presence of a visible crystalline structure on the radiograph of the specimen argues in favour of a benign breast lesion or of a lobular carcinoma in situ. PMID- 3026083 TI - Prostate-specific acid phosphatase in carcinoid tumors. AB - Although prostate-specific acid phosphatase (PASP) has been recognized as a specific marker of tissue of prostatic origin, several investigators have pointed out that some of the carcinoid tumours and islet cell tumours of the pancreas reacted immunohistochemically to PSAP. We investigated 50 cases immunohistochemically comprising 44 carcinoids of the G-I tract, 3 of the bronchus, 1 each of the ovary, kidney and middle ear. PSAP positive cases were, 30 in G-I tract, one each in ovary and kidney. Eighty percent of tumours of hindgut origin were positive. Apart from the immunohistochemical study, the content of PSAP in preoperative serum and tumour tissue was estimated in a case with a rectal carcinoid. Extremely elevated PSAP was confirmed in both the serum and tumour tissue. Neuroendocrine tumours such as pheochromocytoma, medullary thyroid carcinoma, and islet cell carcinoma were investigated as controls. No cells immunoreactive to PSAP were observed in these control cases. Prostate specific antigen was definitely negative in carcinoids. We would emphasize that PSAP may be an excellent marker of carcinoids especially when derived from hindgut. PMID- 3026084 TI - Immunohistochemical examination of lung cancers using monoclonal antibodies reacting with sialosylated Lewisx and sialosylated Lewisa. AB - In order to explore the relationship between the expression of cancer-associated glycolipids such as sialylated Lex (SLEX) and sialylated Lea (SLEA) and the histological subtypes of lung cancers, 30 cases of small cell carcinoma (SCC) and 47 cases of non-small cell carcinoma (non-SCC) were examined immunohistochemically using monoclonal antibodies reacting with SLEX and SLEA. The forty-seven cases of non-SCC included 20 cases of adenocarcinoma, 20 of squamous cell carcinoma and 7 of large cell carcinoma. Tumour cells of most non SCCs expressed SLEX and SLEA. In adenocarcinomas, the number of tumour cells having SLEX and SLEA was more than that of squamous cell carcinomas, large cell carcinomas and SCC. In SCC, 14 of the 30 cases were found to be positive for both antigens. Although the cancer cells of 11 cases of 17 intermediate cell type SCC had both antigens, the cells of only 3 of 13 oat cell tumours expressed SLEX and SLEA. The present study shows that SLEX and SLEA are useful markers for lung adenocarcinomas, that most cases of intermediate cell type of SCCs have characteristics similar to non-SCC but that many oat cell tumours lack them. PMID- 3026085 TI - The influence of cone adaptation upon rod mediated flicker. AB - The influence of annular fields on sensitivity to sinusoidal flicker was assessed in the dark adapted parafoveal retina. Test stimuli were 2 degrees 20' in diameter; annuli had a 2 degrees 20' inner and 7 degrees 30' outer diameter. Rod flicker was studied with a "green" stimulus too dim to influence cones. Selective cone flicker was obtained using red and green flicker in counterphase and yoked together in modulation depth and scotopic illuminance. Results showed the following. (1) Annular stimulation of rods slightly facilitated rod-mediated flicker sensitivity to frequencies less than 10 Hz. In contrast, annular stimulation of cones greatly facilitated rod-mediated sensitivity, particularly for flicker frequencies greater than 7 Hz. We designate this effect, cone-rod interaction. (2) Annular stimulation of cones has a negligible influence upon sensitivity to cone-mediated flicker frequencies less than 15 Hz. In contrast, annular stimulation of rods has a large influence upon sensitivity to cone mediated flicker, an effect we designate rod-cone interaction. (3) Within limits, both rod-cone and cone-rod interaction increase as the annular illuminance increases and as flicker frequency increases; the limiting frequency and illuminance values, however, are different for the two forms of interaction. Results are compared with prior evidence that rod and cone signals summate to produce an absolute threshold or flicker sensation. We suggest that there are at least three mechanisms for interaction between rod- and cone-related signals. PMID- 3026086 TI - Use of selected preventive care procedures. United States, 1982. PMID- 3026087 TI - [Nasal cylindroma]. PMID- 3026088 TI - [Myocardial scintigraphy with 99mTc-pyrophosphate]. PMID- 3026089 TI - [Radioisotope study of the reticuloendothelial cell function of the liver in chronic bronchitis]. PMID- 3026090 TI - [Function of the hypothalamo-hypophyseal-adrenocortical system in stomach cancer patients]. PMID- 3026091 TI - [Treatment of deep caries with a paste containing ethonium, dimexide and aerosil]. PMID- 3026092 TI - [In vitro transcription of human influenza and parainfluenza viruses and its regulation]. AB - The effect of some factors on in vitro transcription of human influenza and parainfluenza viruses was studied. Dinucleotide AfG was shown to stimulate transcription of RNP of human parainfluenza type 3 virus and Sendai virus in the presence of magnesium but not manganese ions same as in the case of influenza viruses transcription. Among two monoclonal antibodies to NP protein of influenza virus, clone F 81 inhibited transcription of RNP of human influenza A viruses, and its influence on transcription of animal influenza viruses was weak. Another clone, H 12, had a low effect. The M protein isolated from influenza and parainfluenza virions inhibited in vitro transcription in the corresponding homologous systems. The inhibiting effect was exerted also by the M protein heterologous to HPIV-3 (M protein of Sendai virus) which suggests the nonspecificity of M protein interaction with transcriptive complexes in large RNA viruses. PMID- 3026093 TI - [Herpes simplex virus-associated antigen in leukemic cells]. AB - Immunological methods (CFT, indirect IF) detected herpes simplex virus-associated antigen in leukocytes of 21 out of 56 leukemia patients. The antigen was more frequently found in leukocytes of patients with chronic lympho- and myeloleukemias, less frequently in acute forms of these diseases. No antigen was detectable in leukocytes of normal donors. PMID- 3026094 TI - [Comparative study of the effectiveness of immunoelectron microscopic methods for detecting rotaviruses and the hepatitis A virus]. AB - Different variants of immune electron microscopy method used for the detection of rotaviruses and hepatitis A virus in specimens from patients were compared. Immune electron microscopy using filtration into agar was shown to be the optimal method for diagnosis of such prevalent infections as rotavirus gastroenteritis and viral hepatitis A. PMID- 3026095 TI - [The level of herd immunity against poliomyelitis in children during the mass vaccinal prophylaxis of this infection]. AB - The results of examinations of blood sera from normal children for the presence of antibody to polioviruses after two courses of mass vaccinations were carried out according to epidemiological indications although differing in duration of the campaign and the range of the child populations involved, are presented. The advantages of simultaneous mass vaccinations covering children from 2 months to 7 years of age have been demonstrated. PMID- 3026096 TI - [Leukocyte migration inhibiting factor in preparations of alpha and gamma interferons]. AB - High titres of leukocyte migration inhibition factor (LMIF) activity were found in native and partially purified preparations of human alpha- and gamma interferon. In the course of chemical purification of interferon up to 98% of LMIF was lost. The highest content of LMIF was in gamma-interferon preparations, while recombinant alpha 2-interferon (reaferon) showed no LMIF activity. PMID- 3026097 TI - [Stimulation of arbovirus reproduction in cell cultures by hormones]. AB - The possibility of stimulation of reproduction and antigen production of arboviruses (tick-borne encephalitis, Japanese encephalitis, western equine encephalomyelitis) in cell cultures (SPEV, chick embryo fibroblasts) by using various hormones (insulin, ACTH, hydrocortisone) and optimal temperatures was demonstrated. The stimulating effect depended upon the type of cell culture, virus, method of cell treatment, and manifested by a significant increase of the hemagglutinating and infectious activity of the replicating viruses. Differences in the structure of populations, biophysical and antigenic properties of the viruses associated with the conditions of their reproduction in vitro were observed. PMID- 3026098 TI - [Use of the agar diffusion precipitation method for detecting rotavirus antigen]. AB - The possibility of rotavirus detection in feces of patients with acute enteric diseases by the agar gel diffusion (AGD) test was studied. The effectiveness of this method was compared with that of the standard method, direct electron microscopy. Both methods showed good correlation of the results, but the AGD test is methodically much simpler which recommends it for diagnosis of rotavirus infection. Rotavirus-specific hyperimmune calf serum may by used as a serological diagnostic preparation for the detection of human rotavirus antigen. PMID- 3026100 TI - [Course of an experimental infection caused by the Langat virus in mice against a background of restricted movement-related stress]. PMID- 3026099 TI - [Immunomorphological methods of studying blood smears in the diagnosis of ophthalmic herpes]. PMID- 3026101 TI - Interactions of vasodilators with calcium entry- and beta-blockers in patients with coronary heart disease. AB - Combinations of antianginal drugs may be used for an additive effect against angina, but also to off-set unwanted effects of one drug with another, either by direct effects or by a reduction of dosage of each drug. Based on earlier studies with separate drugs we have now examined the effect of 150 mg bupranolol combined with 40 mg isosorbide dinitrate (ISDN) in one retarded tablet, given twice daily. 22 patients with CHD entered the study, 11 of those with and 11 without signs of ischemia during exercise. In an acute radionuclide ventriculographic (RNV) study 2 h after the tablet, ejection fraction (EF) during exercise increased only in patients with exercise ischemia (+6%, p less than 0.001). In the other patients EF did not change. After 21 days of treatment echocardiographically determined end-systolic and end-diastolic diameters decreased, resulting in an increase of shortening fraction by 15.6% (p less than 0.05). Heart rate, systolic and diastolic pressure and ST-segment depression decreased significantly. In another acute RNV study the effect of a venous vasodilator, molsidomine 4 mg s.l., was examined after nifedipine 10 mg s.l. in 19 patients with CHD, 9 with and 10 without exercise ischemia. Differences between drugs were most prominent during exercise. In the nonischemic group EF rose by 6.6% after nifedipine (n.s.) and by 14% after molsidomine (p less than 0.01 against control). In the group with ischemia EF rose by 12.6% after nifedipine and by 17.4% after additional molsidomine, significant against control (p less than 0.01) as well as against nifedipine (p less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3026102 TI - Mode of action of nitrates at the cellular level. AB - Nitrates probably induce vasorelaxation via a rise of cytosolic cGMP, and subsequent phosphorylation of target proteins by cGMP-dependent protein kinase. A dual type of action by this mechanism seems likely: cGMP-dependent protein kinase relaxes chemically skinned vascular smooth muscle which has no functioning cell membrane. Thus, the contractile apparatus with its regulatory and contractile proteins may be one of the targets for their action. Calcium visualization techniques using aequorin or quin-2, and ion flux studies showing suppression of Ca2+-dependent 86Rb efflux by nitrates and 8-Br-cGMP suggest that the cytosolic calcium level is another target for their action. Whether this lowering of intracellular calcium occurs via cGMP-dependent activation of the sarcolemmal Ca2+ extrusion ATPase, requires confirmation. PMID- 3026103 TI - Experimental studies on the mechanism of glyceryl trinitrate-tolerance and dependence. AB - Tolerance to glyceryl-trinitrate (GTN) in arteries and veins was studied in different in vitro and in vivo models. The development of GTN tolerance was associated with a down-regulation of the cGMP system, i.e. an impairment of the nitrocompound-dependent activation of guanylate cyclase and an increase of the cGMP-phosphodiesterase activity. We suggest that a decreased function of the cGMP system might cause an impairment of the negative feedback loop involving cGMP in blood vessels, and a concomitant supersensitivity to contractile stimuli. PMID- 3026104 TI - Synthetic peptide vaccines against foot and mouth disease. AB - Synthetic peptide vaccines against foot-and-mouth disease (FMD) have been under examination for several years. Recent reports indicate that effective synthetic vaccines for use in cattle are being developed. However, there are still some unresolved questions regarding the response of cattle and other animals to these vaccines. This brief review describes some recent studies of synthetic vaccines against FMD and draws attention to some deficiencies in our understanding of the response to these vaccines. PMID- 3026105 TI - Prospects for the clinical management of human cytomegalovirus infections. AB - Infection of susceptible populations by human cytomegalovirus (HCMV) is a significant public health problem in Western societies. Vaccination with live attenuated strains of HCMV has demonstrated some degree of clinical benefit but objections based on the possibility of these viruses becoming latent and their potential oncogenicity must be considered. Our knowledge of the biology and immunology of HCMV, although advancing rapidly, is still a long way short of being able to predict candidate subunit vaccines based on virus encoded proteins or glycoproteins. Treatment of the disease by injection of antibodies awaits a breakthrough and chemicals effective in the control of other human herpes viruses are disappointingly ineffective against HCMV. Clearly, prophylaxis is preferable to therapy and it is in the design of new effective vaccines that endeavours must be channelled so that we can control complications associated with severe clinical infection with this virus. PMID- 3026107 TI - Efficacy of vaccination with Skinner vaccine towards the prevention of herpes simplex virus induced cervical carcinoma in an experimental mouse model. AB - The incidence of cervical carcinoma, which had been induced by vaginal application of inactivated herpes simplex virus type 2 (HSV-2) with 20% croton oil, was significantly reduced in mice prevaccinated with the Skinner herpes vaccine. There was evidence of an immunological response in both vaccinated and unvaccinated mice. PMID- 3026106 TI - Combined vaccination with live oral polio vaccine and the bovine rotavirus RIT 4237 strain. AB - A clinical trial was carried out in 3-month-old infants to assess whether concomitant oral administration of live polio and rotavirus RIT 4237 vaccines would reduce their immunogenicities as a result of mutual interference. One hundred and sixty breast-fed male and female infants were randomly allocated to four study groups to receive in a blind fashion the poliovirus vaccine, the RIT 4237 vaccine, a combination of both vaccines or a placebo preparation. Antibody titres were measured in pre- and postvaccination serum samples by the ELISA test and the neutralizing antibody test (NT) for rotavirus and by the NT for polioviruses types 1 and 3. The percentage of subjects with immune responses to rotavirus in the placebo group was low, indicating the absence of wild rotavirus circulation in the population. Antibody responses against polio types 1 and 3 were found in about a quarter of the infants receiving a placebo because the study was performed during a polio vaccination campaign when vaccine viruses are known to circulate. The results showed that 73% of seroconversion was obtained when RIT 4237 was administered alone and that the responses to polioviruses types 1 and 3 were good. However, simultaneous administration of polio and RIT 4237 vaccines caused a significant reduction of the antibody response to rotavirus but not to polioviruses types 1 and 3. PMID- 3026108 TI - [Regulation of steroid hormone levels in stress in old age (component analysis)]. PMID- 3026109 TI - Further characterization of a morphogenetic mutant of the foot-and-mouth disease virus. AB - In this paper we describe further characterization of a foot-and-mouth disease virus (FMDV) temperature-sensitive mutant, ts 139. This mutant was very sensitive to heat inactivation, suggesting that its viral particles are somehow altered. The electrophoretic analysis of ts 139 structural proteins indicated that VP2 has an altered mobility. Furthermore, two known protein precursors of VP2, VP0 and p88, were shown to be altered, as was p64, which supports a VP2 precursor role for p64. The ts 139 viral particle assembly pathway was analyzed during viral replication. The empty-capsid to complete-viral-particle ratio was clearly increased compared to that found for the wt strain, indicating an alteration in the morphogenetic process. This encouraged us to search for the presence of possible viral precursors of low sedimentation coefficient which have not been described previously in aphthovirus-infected cells. Analysis of the viral morphogenetic process in ts 139-infected cells demonstrated the presence of viral complexes formed by VP0, VP1 and VP3 which sedimented slightly faster than a 12 S marker. The protein composition, the sedimentation coefficient of this complex and the pulse-chase results strongly suggest that it is a morphogenetic precursor of the mature viral particle. PMID- 3026110 TI - Antigenic comparison of foot-and-mouth disease virus serotypes with monoclonal antibodies. AB - The capsid structures of the 7 serotypes of foot-and-mouth disease virus have been compared utilizing a series of neutralizing monoclonal antibodies which were previously shown to recognize at least 4 distinct epitopes on type A12 virus. A radioimmune binding assay using radioactively labeled antigens and the monoclonal antibodies revealed that certain conformation dependent epitopes are conserved among A subtypes, while some continuous epitopes are conserved among A subtypes as well as other FMDV serotypes. On the basis of differential reactivity among other FMDV subtypes and serotypes two additional epitopes have been defined on the A12 particle. Binding and neutralization assays revealed that the presence and function of an epitope are not necessarily correlated. PMID- 3026111 TI - The genome of bovine herpesvirus 1 (BHV-1) strains exhibiting a neuropathogenic potential compared to known BHV-1 strains by restriction site mapping and cross hybridization. AB - Bovine herpesvirus 1 (BHV-1) strains can be differentiated by their DNA and polypeptide patterns, and by antigenic properties as demonstrated by monoclonal antibodies. We classified the BHV-1 strains according to these data as BHV-1.1, BHV-1.2 (a/b) and BHV-1.3 (a/b). BHV-1.1 and BHV-1.2 correspond to the well known 'common' BHV-1 strains, whereas BHV-1.3 has only recently been recognized and exhibits a neuropathogenic potential. In the present paper we describe the structural genome characteristics of BHV-1.3 compared to those of the other BHV-1 strains, examined by means of restriction site mapping, electron microscopy and cross-hybridization. Our results also confirm and complete data concerning BHV 1.1 and BHV-1.2 published by other authors. The following main conclusions can be drawn from our investigations: BHV-1.1 and BHV-1.2 differences are restricted to distinct genomic regions, characterized by loss or gain of restriction sites. BHV 1.3, however, differs from the other BHV-1 strains in restriction site alterations throughout the whole genome. Electron microscopy showed the typical BHV-1 DNA structure for BHV-1.3. Genetic homology between BHV-1.1 and BHV-1.2, reported to be about 95%, was confirmed by cross-hybridization, and a similar high base sequence homology for BHV-1.3 could be shown. PMID- 3026112 TI - Temperature-dependent cytocidal effects of Moloney murine sarcoma virus. AB - Infection of L6E9 rat myoblasts with Moloney murine sarcoma virus (Mo-MuSV) at 37 degrees C resulted in the killing of target cells in less than one week. However, cells infected at 37 degrees C and shifted to 40 degrees C were not killed but underwent morphological transformation and had reduced levels of virus-specific RNA. A marked decrease in a cellular 55 kDa protein, as judged by immunoblotting, correlated with increased expression of Mo-MuSV RNA in infected cells. PMID- 3026113 TI - Localization and characterization of Sendai virus nonstructural C and C' proteins by antibodies against synthetic peptides. AB - Antibodies were raised in rabbits against two synthetic peptides, each 30 residues in length, one corresponding to the predicted common carboxyl termini of the nonstructural C and C' proteins of Sendai virus and the other to the unique amino terminus of the larger C protein. Each peptide was inoculated as a covalent complex with tetanus toxoid or in uncomplexed form. Only antibodies to the free carboxyl-terminal peptide precipitated both C and C' proteins made by in vitro translation of viral mRNA and reacted with the C protein from infected cells. These results confirm that the C and C' proteins are carboxyl-coterminal. Contrasting with the reported colocalization of intracellular measles virus C proteins with nucleocapsid inclusions, immunofluorescence studies revealed that Sendai virus C proteins were uniformly distributed in the cytoplasm whereas the viral P protein was present in inclusions that were mainly perinuclear. Since almost all P protein molecules are associated with viral nucleocapsids, these observations suggested that Sendai virus C protein molecules may be both nucleocapsid-associated and free in the cytoplasm. This interpretation was supported when the C and C' proteins were found in both nucleocapsid and free protein fractions of cell lysates. Anti-C antibodies did not inhibit viral RNA synthesis when added to an extract of infected cells. This result was consistent with the conclusion that the C proteins have no direct role in viral transcription, since virions lack C proteins but are transcriptionally active. Therefore, the functions of the C proteins remain undefined. PMID- 3026114 TI - Duplication of a viral enhancer sequence improves the stability of a vector based on BPV-1 DNA. AB - Various recombinant constructions involving bovine papillomavirus type 1 (BPV-1) DNA and bacterial plasmids have been tested for their ability to transform mouse C127 cells and replicate as intact extrachromosomal monomeric structures. When BPV-1 DNA was linked to pBR328, pAT153 or derivatives of these plasmids lacking the 344 bp HindIII-BamHI fragment or another small segment, the resulting vectors replicated in C127 cells as high molecular weight structures and, in some cases, deleted extrachromosomal forms. The sequences which became deleted were generally the non-BPV-1 sequences. Duplication of the 3' distal enhancer sequence of BPV-1 DNA in one of the vectors increased its stability upon introduction into C127 cells, but some deleted and high molecular weight forms were still observed. PMID- 3026115 TI - Induction of mutations in bacteria by a fragment of DNA from herpes simplex virus type 1. AB - A bacterial assay was developed for the study of mutagenesis by DNA of herpes simplex viruses. The histidine mutations from two of the Ames mutagenesis tester strains were recombined into the Salmonella histidine operon of the F'8 plasmid and each was transferred to a derivative strain of E. coli C from which the resident histidine operon had been deleted. One tester strain could be reverted by a chemical mutagen which induces frameshift mutations and the other could be reverted by a mutagen which induces base-pair substitution mutations. The BamHI G fragment of herpes simplex virus type 1 was cloned in each orientation into the BamHI site of the expression vectors pUC7, pUC8 and pUC9 and were introduced into the new strains of E. coli. The pUC9 plasmid carrying the BamHI G fragment of herpes simplex virus type 1 with the G-E' site closest to the lac promoter showed a higher rate of reversion in the frameshift strain, which varied up to 39-fold greater than the background rate. Since many mutagens are carcinogenic these data suggest the existence of a mutagenic peptide of herpes simplex virus type 1 which might be involved in cell transformation. PMID- 3026116 TI - [Changes in pyramidal tract response to direct stimulation of the sensomotor cortex of the rabbit during formation of a conditioned reflex]. AB - Changes of pyramidal tract (PT) response were analyzed, reflecting the final result of cortical interaction in the process of combinations of direct stimulations of cortical surface in nonimmobilized and unanaesthetized rabbits. It has been shown that in a situation, modelling conditioning, changes take place of the first direct component--the D-component--of the PT response (reflecting the excitability of the PT neurones), as well as changes of the successive indirect synaptic component--I-component (reflecting the excitability of presynaptic cortical elements and of intracortical synaptic connections). I component changes were significantly more expressed. In most cases the I component of the response was increased. The obtained data testify to an increase of synaptic efficiency in the process of temporary connection formation and to possible change (increase or reduction) of excitability of PT neurones. PMID- 3026117 TI - [Cholinergic regulation of the activity of somatosensory cortex neurons during early postnatal ontogeny of the cat]. AB - In 11-14 days kittens, about 20% of neurones in the somatosensory cortical zone react to stimulation of subpallidal region which is a source of cholinergic projections to the cerebral cortex. The effect of subpallidal region stimulation is reproduced in case of microiontophoretic acetylcholine application and blocked by atropine what points to its cholinergic nature. Cholinergic stimulation causes inhibition of the background and evoked activities of the cortical neurones while, as it is known, in adult cats, acetylcholine mainly stimulates a reaction of activation. It is postulated that in kittens at the end of the second week of postnatal development, cholinergic innervation of the cortex significantly differs from the definitive one by its quantitative and functional parameters. PMID- 3026119 TI - [Autopsy findings in patients with acquired immunodeficiency syndrome]. AB - We have evaluated the autopsies of 11 patients with HTLV III/LAV-infection. The clinical diagnosis was AIDS in 10 cases and AIDS related complex (ARC) in one case. The most common infectious disease was pneumocystis carinii pneumonia, occurring in 5 cases. 3 patients showed evidence of mycobacterial infections and another three showed cytomegalovirus infections. Kaposi's sarcoma was found in 4 and other malignancies in 3 cases. Our results are in agreement with the findings of other authors. PMID- 3026118 TI - [Morphometric analysis of fibrocystic breast disease]. AB - We used the automated microscopic image analysis (system "Robotron A 6471"; software AMBA/R) to study the ductal epithelium in biopsies from 88 patients with fibrocystic breast disease, including 52 with proliferative mastopathy. Significant differences were demonstrated using reproducible data and selective caryo- and histometric parameters which allowed the separation of ductal hyperplasia and ductal cell proliferation with atypia. In addition, multiparameter analysis indicated that cases with proliferative mastopathy III can be separated into two groups which have probably a different high risk with respect to carcinoma. PMID- 3026120 TI - [Bone substitutes. Hydroxylapatite]. PMID- 3026121 TI - Establishment of a continuous cell line derived from leukaemic cattle. PMID- 3026122 TI - Experimental infection of red fox with canine parvovirus. PMID- 3026123 TI - Characterization of a sheep pox virus attenuated in cell culture. PMID- 3026124 TI - [Detection of modification-restriction systems in Yersinia enterocolitica]. AB - Four Y. enterocolitica strains (10166, 10373, 2119, 5513) have been studied for the presence of the enzymatic systems of modification-restriction (M-R). As revealed with the use of cross titration, strains 10166 and 10373 contain M-R systems, supposedly of type II. PMID- 3026125 TI - [Choice of the method for isolating conjugates of staphylococcal protein A with peroxidase for immunoenzyme analysis]. AB - An attempt to obtain the preparations of peroxidase-labeled staphylococcal protein A, intended for use in the enzyme immunoassay, by the glutaraldehyde method has failed. The modified periodate method permitting the preparation of active conjugates of staphylococcal protein A with peroxidase has been developed. PMID- 3026126 TI - [Immunohematologic changes in pituitary adenomas]. AB - The authors conducted studies of colony-producing capacity of precursor cells of bone marrow granulomonopoiesis and cytogenetic study of blood T lymphocytes in 13 patients with adenoma of the hypophysis. It was established that adenomas of the hypophysis are attended by the development of functional deficiency of T lymphocytes and the resultant disturbance of T cell regulation of granulomonopoiesis. The disorder of granulomonopoiesis occurs at the level of the precursor cells in the bone marrow. PMID- 3026127 TI - Defective cAMP metabolism and defective memory in Drosophila. PMID- 3026128 TI - Protein phosphorylation in dunce memory-mutant Drosophila. PMID- 3026129 TI - The role of DTNB light chain in the contractile properties of skeletal muscle. AB - The mechanical and molecular dynamical properties of the glycerinated muscle fibres from m. psoas of rabbit were investigated after treatment with 5,5' dithiobis/2-nitrobenzoate (DTNB). After DTNB treatment the fibre bundles shortened and exerted force in ATP-relaxing solution. The rate of the ATP hydrolysis in relaxation-inducing medium increased significantly after DTNB treatment, and simultaneously the amount of the LC2(DTNB) light chain decreased to 40 per cent of the original value measured by gel electrophoresis. The electron paramagnetic resonance spectra of muscle fibres labelled with maleimide spin label after DTNB treatment exhibited high degree of order of label in rigor, but this order decreased after shortening. The experiments support the assumption that, in vertebrate skeletal muscles, besides the troponin-tropomyosin regulation system which developed later in evolution, the LC2 light chain should maintain some regulatory properties. PMID- 3026130 TI - [Prognostic factors in breast carcinoma]. AB - The prognostic factors in breast cancer are numerous and interdependent. The single most important variable is the total tumor cell burden as well as the marked heterogeneity of tumor cell population. In women with clinical localized disease, the progressive increase in the diameter of primary tumor is associated with higher incidence of histologically involved axillary nodes; in turn, this involvement, and particolarly its extent, represents an indirect sign of distant micrometastases. In women with clinical advanced disease, the extent of tumor cell burden (inflammatory carcinoma, multivisceral involvement) is associated with a decrease in the response rate by drug therapy. The percent of drug resistant neoplastic cells is inversely related to the tumor mass; this resistant fraction represents today the limiting factor to a more effective contribution of medical treatment (i.e. long-term complete remission and cure) in all stages of breast cancer. The concept of breast cancer as systemic disease in a higher percent of women than thought in the past, allows to perform breast-saving procedures without affecting prognosis and stimulates treatment strategies utilizing a combined modality approach for high-risk women. PMID- 3026131 TI - [Multisystemic viral infection and juvenile diabetes (type I). Apropos of a clinical case]. PMID- 3026132 TI - Primary malignant lymphoma of the brain: an immunohistochemical study of eight cases using a panel of monoclonal and heterologous antibodies. AB - Frozen sections of eight primary malignant lymphomas of the brain were examined by the immunohistochemical methods using a panel of monoclonal and heterologous antibodies to B lymphocyte (immunoglobulins, BA-1, Leu-12 and HLA-DR), T lymphocyte (OKT-11 and Leu-1) and monocyte (OKM-1) surface markers. Paraffin sections were also used in the examination of cytoplasmic immunoglobulins. Surface and/or cytoplasmic immunoglobulins (Ig) were observed in seven cases, four of which were shown to be distinctly monoclonal and the other three less so. The remaining 1 case showed no distinct staining for Ig. BA-1, Leu-12 and HLA-DR stainings were positive in four, four and five cases, respectively. The marker phenotypes of (BA-1, Leu-12, HLA-DR) were shown to be (+, +, +) in one lymphoma, (+, -, -) in three, (-, +, +,) in three, and (-, -, +) in one. Thus, it was demonstrated that the present lymphoma cases showed a marked immunological heterogeneity, and it was shown that all of them including the Ig-negative case revealed one or more of these three additional B cell markers, indicating B cell lineage of these cases. Examination of T cell and monocyte markers revealed positive staining in normal or reactive lymphoid cells distributed around blood vessels or sporadically in tumor tissues, but not in lymphoma cells. Epstein-Barr virus (EBV)-associated nuclear antigen was not demonstrated in the seven cases examined, making it unlikely that these lymphomas were related with EBV infection. PMID- 3026133 TI - Proliferating potential of folliculo-stellate cells in human pituitary adenomas. Immunohistochemical and electron microscopic analysis. AB - Folliculo-stellate cells (FS cells) in 40 pituitary adenomas and portions of anterior pituitary adjacent to the tumor in 26 cases were investigated immunohistochemically, using polyclonal antisera to S-100 protein (S-100) and glial fibrillary acidic protein (GFAP). The objective was to clarify the histological behavior of the FS cells. In most pituitary adenomas there were few or no S-100- or GFAP-positive cells, in comparison with numerous positive cells in the parts of the adenohypophyses compressed by adenomas. However, positive FS cells were observed in some types of pituitary adenomas. Growth hormone and prolactin producing adenomas frequently contained significant amounts of FS cells. In non-functioning adenomas, an unique case of FS cell adenoma was present. The adenoma was composed mainly of FS cells and immature glandular cells. The FS cells were sometimes located around follicles containing Periodic acid Schiff-positive material. Therefore, the FS cell adenoma is characterized by S-100- and GFAP-positive FS cells and PAS-positive follicles. In this type of adenoma, FS cells seemed to be the main proliferating component. In parts of the adenohypophyses adjacent to the adenomas, GFAP-positive FS cells were numerous. In the pathological conditions FS cells may possess the potential of reactive proliferation. PMID- 3026134 TI - Proliferation of Schwann cells in demyelinated rat sciatic nerve. AB - Experimental demyelination was induced by intraneural injection of anti galactocerebroside serum into the sciatic nerves of rats. Schwann cells undergoing mitotic division were observed between days 3 to 9 after the injection and demyelinated segments were still associated with macrophages. Dividing Schwann cells were often present in association with both unmyelinated and myelinated fibers. Whether or not, daughter Schwann cells migrate along the same fiber towards neighboring demyelinated segments remains unclear. When Schwann cells attached to axon membranes of demyelinated segments were studied at later time points, they were present in clusters randomly at various regions of the segments. There was no proximo-distal gradient for the wave of Schwann cell proliferation. Mean Schwann cell internuclear distances were around 40-50 microns at the earliest time of remyelination. Schwann cell redistribution and remyelination progressed regardless of the length of demyelinated segments. PMID- 3026136 TI - Progressive supranuclear palsy with Lewy bodies. AB - An autopsy case is reported which revealed not only clinical and neuropathological features of progressive supranuclear palsy, but also the presence of large numbers of Lewy bodies in the brain stem nuclei and cerebral cortex. This case seems to be progressive supranuclear palsy with Lewy bodies distributed as in Parkinson's disease. Such case has not been previously reported. PMID- 3026135 TI - Intercellular deposits of basement membrane material in active human pituitary adenomas detected by immunostaining for laminin and electron microscopy. AB - Thirty-eight human pituitary adenomas (24 endocrine active and 14 endocrine inactive tumors) were studied immunohistochemically for the presence of the basement membrane component, laminin, and ultrastructurally for the presence of basement membrane. Immunoreactivity of laminin delineated staining of epithelial and endothelial basement membranes, the reaction product being confined mostly to the perivascular zones. Moreover, a hitherto undescribed presence of intercellular laminin-positive droplets was observed in ten of the active adenomas (nine patients with hyperprolactinemia and/or acromegalia and one patient with Cushing's syndrome). Concurrently, at the ultrastructural level, bunches of basement membrane-like material intermingled between the adenoma cells were demonstrated in seven of these ten active adenomas. Furthermore, secretory granules were entrapped occasionally in this intercellular matrix, indicating a mutual dependence between excessive hormone extrusion and an increase of "misplaced" deposits of basement membrane components, e.g., laminin. PMID- 3026137 TI - Suppressive effect of monocytes in vitro in patients with carcinoma of the uterine cervix. AB - The adherent cell fraction (AdC) of human peripheral blood mononuclear cells (PBM) contains two cell types of opposing function in vitro. Dendritic cells (DC) act as antigen-presenting cells (APC) in vitro, while monocytes (Mo) have a suppressive effect on antigen activation of T cells. In this report we show that patients with cervical carcinoma have a significantly increased number of suppressive Mo compared with healthy controls. The T cell response to Herpes Simplex Virus (HSV-1) and PPD was about the same in the two groups, but after removal of Mo by adherence a significantly higher T cell response was seen among the patients with advanced stage (Figo stage IIb-IVa) compared with patients in early stage (Figo stage Ia-IIa) and healthy controls. These observations indicate that patients with cervical carcinoma have an increased number of T cells reactive with HSV, and normal DC function. The relative suppression expressed on a per Mo basis was the same in all groups, which indicates that the increased suppression in the patients was caused by an increased number of Mo, and not by changes in their activation state. PMID- 3026138 TI - Immunohistochemical study on the distribution and significance of mononuclear cells in human breast carcinoma. AB - Lymphocyte subsets in the tumor nests of breast carcinoma were immunohistochemically investigated and a quantitative analysis was added. The majority of cases showed predominance of T cell and suppressor T cell (T8). A decrease in number of lymphocyte subsets and the helper T (T4)/T8 ratio in the stroma of tumor nests correlated well with the progression of clinical stage and the presence of metastasis. This correlation could not be found in the peripheral region of the tumor nests. Macrophages and NK cells were infrequently observed only in the peripheral region of ductal carcinoma. T cell infiltration was prominent in medullary carcinoma with lymphocyte infiltration (MC), and macrophages, NK cells, and T zone histiocytes were frequently encountered. For the purpose of knowing the activity of T cells, IL-2 receptor (Tac) and transferrin receptor were examined immunohistochemically. The fact that a few activated T cells were found only in the peripheral region of tumor nest suggested the local immune response in ductal carcinoma not to be so active as to reject the tumor cells. Since numerous activated T cells were recognized in the tumor nests of MC, this type of breast carcinoma was thought to have a higher immune reactivity. There was little evidence indicating NK cells to play a role for natural cytotoxicity in breast carcinoma. PMID- 3026139 TI - Giant cell tumor of tendon sheath. An immunohistochemical study of 28 cases. AB - An immunohistochemical study was carried out on 28 cases of giant cell tumor of tendon sheath. Although this tumor has been considered to be of histiocytic origin on the basis of light and electron microscopic findings, there remains some debate about the histogenesis of the tumor. To clarify this point, by using the PAP method, each surgical specimen was stained for alpha 1-antitrypsin, alpha 1-antichymotrypsin, lysozyme, ferritin, neuron specific enolase, and S-100 protein. Tumor cells in fifteen out of 28 cases were positively stained for alpha 1-antitrypsin, 19 for alpha 1-antichymotrypsin, 23 for lysozyme, 22 for ferritin, 22 for neuron specific enolase, but no case for S-100 protein. These results suggest that this tumor is composed of cells with histiocytic character. In addition, from the immunohistochemical point of view, at least two types of giant cells seem to exist in this disease. PMID- 3026140 TI - Activation of cytomegalovirus infection in immunosuppressed cynomolgus monkeys inoculated with varicella-zoster virus. AB - A systemic activated cytomegalovirus (CMV) infection was fortuitously detected in almost all monkeys which had been immunosuppressed with antithymocyte globulin (ATG), cyclophosphamide (CY), and cortisone acetate (CS) before and after experimental inoculation with varicella-zoster virus (VZV). They developed exudative pneumonia, and the lesions in visceral organs and tissues contained cytomegalic cells with intranuclear inclusion bodies, in which viral antigens, specific for CMV, but not inoculated VZV, were detected by immunofluorescence. Serological study of paired sera from these monkeys ascertained preexisting CMV infection. Under the present experimental conditions, this infection was highly reproducible and always occurred within three, but not two, weeks of immunosuppression in monkeys inoculated with VZV. We therefore examined the host factors involved in activation of latent CMV. The immunocompetence of the host was destroyed almost completely with treatment of ATG, CY, and CS, but not with combinations of two of these agents, revealing the systemic depletion of lymphoid cells in tissues including the thymus medulla. Although the role of VZV in the induction of CMV remains uncertain, the heterologous VZV inoculum may have produced some effects equivalent to the allogeneic reaction to release latent CMV. These monkeys may represent an animal model of "opportunistic" CMV infection in immunocompromised and/or allografted humans. PMID- 3026142 TI - Abnormalities of sympathetic regulation after cervical cord lesions. AB - Microneurographic recordings suggest that hypertensive reactions in patients with cervical spinal cord lesions usually are due, not to exaggerated sympathetic nerve reactions, but rather to the combination of more diffuse sympathetic nerve activation than normally, exaggerated vascular reactions and absence of blood pressure restraining reflexes. PMID- 3026141 TI - Computed tomography in early stages of testicular carcinoma. Size of normal retroperitoneal lymph nodes and lymph nodes in patients with metastases in stage II A. A SWENOTECA study: Swedish-Norwegian Testicular Cancer Project. AB - From the SWENOTECA Project, the CT findings in 156 patients treated by bilateral retroperitoneal lymphadenectomy were reviewed. Of these, 112 were in stage I (no metastases) and 44 in stage II A (metastases in normal-sized lymph nodes and in nodes with a maximum diameter in the transverse plane of less than or equal to 20 mm). The normal size of lymph nodes in young Scandinavian men was found to be less than 10 mm X 8 mm above the bifurcation, except in the area below the left renal vein to the left of the aortic midline where the normal size was found to be maximally 14 mm X 10 mm. The addition of a lymphangiographic contrast medium did not change the size of the lymph nodes significantly above the bifurcation, while changes of importance were noticed in the pelvic area. Normal size without contrast medium was found to be at the most 15 mm X 10 mm and after addition of contrast medium to the nodes 28 mm X 12 mm. The results of the CT findings in the stage II A group were not impressive but changed somewhat for the better using the new limits concerning size of retroperitoneal lymph nodes. The impact of using different limits is discussed and it is concluded that metastases in normal sized or almost normal-sized lymph nodes will continue to be a diagnostic problem, at least when using CT. PMID- 3026144 TI - Inactivation of metalloenzymes by lysinoalanine, phenylethylaminoalanine, alkali treated food proteins, and sulfur amino acids. AB - Synthetic lysinoalanine (LAL) may be a more effective inhibitor of the zinc containing enzyme carboxypeptidase A than is ethylenediamine tetraacetic acid (EDTA). The enzyme is also inactivated by alkali-treated, lysinoalanine containing food proteins such as casein, high-lysine corn protein, lactalbumin, soy protein isolate, and wheat gluten, and by alkali-treated zein, which contains no lysinoalanine. Zinc sulfate regenerates only part of the enzymatic activity after exposure to the treated proteins. The extent of inhibition increases with protein concentration and time of treatment. Any inhibition due to phytate is distinct from that due to the treatment. Phenylethylaminoalanine (PEAA), derived from biogenic phenylethylamine, inhibited enzymatic activity of the metalloenzyme carboxypeptidase A (CPA). The inhibition was maximal at pH 7.0 in the pH range 7 to 8.5. The extent of inhibition increased with time of treatment and PEAA concentration. N-acetyl-PEAA did not inhibit the enzyme, suggesting that the free alpha-NH2 group is required for inhibition. PEAA, LAL, sodium phytate, and cysteine also inactivated the copper enzyme, polyphenol, oxidase (tyrosinase) which plays a major role in enzymatic (oxidative) browning of foods. Analogous comparative studies with LAL, EDTA, and sodium phytate suggest that the potency of PEAA as an inhibitor of CPA is similar to that of sodium phytate, and that of the four compounds tested, PEAA is least effective against tyrosinase. Related studies of the iron and copper containing enzyme cytochrome C oxidase showed that EDTA was not inhibitory, PEAA was slightly inhibitory, and LAL and sodium phytate were stronger inhibitors. Mechanistic explanations are offered to account for some of these observations. The possible relevance of these findings to in vivo protein digestion, enzymatic (oxidative) browning of foods, and the mechanism of the lysinoalanine effect on kidney cells are also discussed. PMID- 3026143 TI - Protease inhibitors: their role as modifiers of carcinogenic processes. PMID- 3026145 TI - Effects of carotid artery compression test on regional cerebral blood volume, hemoglobin oxygen saturation and cytochrome-C-oxidase redox level in cerebrovascular patients. PMID- 3026146 TI - Kinetic analysis of cardiac beta-receptors in perfused working rabbit hearts. AB - Myocardial adrenergic beta-receptors were isolated and partially purified from the nonischemic perfused working rabbit hearts. Using highly radioactive beta receptor antagonist, dihydroalprenolol (DHA), properties of beta-receptors were investigated by kinetic equilibrium analysis when the physiological function of the heart appeared to be normal. At the concentration of 10 nM DHA dissociation constant (Kd) was 14.9 nM and there were at least two distinctly different DHA binding sites, based on the analysis of the dissociation rate of DHA-receptor complex. Identification of the two distinctly different DHA binding sites was not obvious from the analysis of Scatchard plot. PMID- 3026148 TI - Near infrared optical monitoring of cat skeletal muscle during hypercapnia. PMID- 3026147 TI - Lowered oxygen supply and calcium (Ca) accumulation in rabbit proximal tubules. PMID- 3026150 TI - Type III hyperlipoproteinemia: a focus on lipoprotein receptor-apolipoprotein E2 interactions. PMID- 3026149 TI - Cellular oxygen utilization and hypoxia: interaction of dithiols with cellular electron transfer systems. PMID- 3026151 TI - Dietary influences on neurotransmission. AB - Diet clearly influences neurotransmission. This can be important in grossly undernourished children. It can also be important in children in whom normal homeostatic mechanisms governing food intake are bypassed. Subtle differences in behavior can occur with physiologic variation in food intake. Components of foods can also be used as drugs. Starvation can impair neuronal maturation and can have lasting effects upon behavior and intellectual performance. The extent of starvation's impact upon the brain depends upon whether undernutrition occurred during a critical phase in brain development. Short-term fasting has small, but significant, effects upon intellectual performance. Even when gross malnutrition is not present, subtle changes in diet may modulate brain function. Tryptophan, tyrosine, and choline in the diet are used as precursors for neuronal synthesis of serotonin, dopamine and norepinephrine, and acetylcholine, respectively. It is likely that the brain's sensitivity to certain components of the diet exists to permit monitoring of food intake by the central nervous system. Tryptophan, tyrosine, and choline may be useful in treatment of humans with sleep disorders, pain depression, mania, hypertension, shock, or dyskinesias. Other components of the diet that may affect behavior include food additives, sugar, and caffeine. Food additives may exacerbate hyperactive symptoms in a small proportion of children with attention deficit disorder. Given that there is little potential for harm and that there is a subpopulation that may respond, a trial of a diet that contains no food additives may be a valid diagnostic approach for children with attention deficit disorder who do not respond to stimulant therapy or for children for whom stimulant therapy is not desired. Refined sugar has been blamed for many behavioral abnormalities. Subtle effects of carbohydrate upon behavior have been reported, but the existing data do not support the hypothesis that sucrose or fructose exert special effects upon neurotransmission. Caffeine is easily detected as a stimulant by humans, but it has little effect upon cognitive function. Administration of large doses of vitamins has no beneficial effect in most humans with schizophrenia, attention deficit disorder, autism, Down's syndrome, or drug addiction. Large doses of niacinamide may even be harmful, as they may cause hepatic damage. PMID- 3026152 TI - Mechanistic aspects of DNA topoisomerases. PMID- 3026154 TI - [Molecular epidemiology of adenoviral conjunctivitis in Sapporo, Japan and Manila, Philippines]. PMID- 3026155 TI - A serological survey of enteric parvovirus infections in Finnish fur-bearing animals. PMID- 3026156 TI - Isolation of a herpesvirus serologically related to bovine herpesvirus 1 from a reindeer (Rangifer tarandus). PMID- 3026157 TI - Cystic masses of the knee: arthrographic and CT evaluation. PMID- 3026153 TI - Leukotrienes in inflammation. PMID- 3026158 TI - Bone scan-positive and radiograph- and CT-negative vertebral lesion in a woman with locally advanced breast cancer. PMID- 3026159 TI - Thoracic manifestations of AIDS. AB - Of 67 hospitalized AIDS patients, 39 had pulmonary pathology. More than half of these patients died of pulmonary disease. Pneumocystis carinii, cytomegalovirus, Cryptococcus neoformans and Mycobacterium avium-intracellulare were the most common pulmonary pathogens, and Kaposi's sarcoma was the most common neoplasm. Infections and neoplasms frequently coexist in the thorax of an AIDS patient. The chest radiograph may be normal in an AIDS patient with active Pneumocystis pneumonia. PMID- 3026160 TI - The effects of cytochrome C on the extent of myocardial infarction and regional function of the ischemic myocardium. AB - The effects of cytochrome C, an electron carrier in the process of oxidative phosphorylation, on infarct size and regional left ventricular function after a coronary artery occlusion were investigated. Thus, in 30 dogs, 1 minute after left anterior descending coronary artery occlusion, 99mTc-labeled albumin microspheres (8 mCi) were injected into the left atrium for subsequent assessment of the hypoperfused zone, that is, the area at risk of infarction. Fifteen minutes after coronary artery occlusion, dogs were randomized into a control group (n = 15) and a cytochrome C-treated group (n = 15). The latter immediately received cytochrome C, 2.5 mg/kg intravenously. Six hours after coronary artery occlusion the dogs were sacrificed and their left ventricles were cut into 3 mm thick slices. Infarct size was determined by triphenyltetrazolium chloride staining and measured by planimetry. The same slices were then submitted to autoradiography and the hypoperfused zone was then measured by planimetry. The hypoperfused zone was 22 +/- 2% and 23 +/- 2% of the left ventricle in the control and treated groups, respectively (NS), indicating that the extent of myocardium at risk before treatment was similar. The extent of the hypoperfused zone which evolved to necrosis was 90 +/- 3% in the control group but only 50 +/- 7% in the treated group (p less than 0.001). Myocardial salvage in the treated group was paralleled by improvement in systolic wall thickness of the ischemic segment as measured by two-dimensional echocardiography. Thus, cytochrome C reduced the extent of myocardial necrosis by 44% and improved systolic function of the ischemic myocardium. PMID- 3026161 TI - Evaluation of air-purifying respirators for protection against toluene diisocyanate vapors. PMID- 3026162 TI - Toxicology of solvents. PMID- 3026163 TI - Man-made vitreous fibers: an overview of studies on their biologic effects. AB - Clinical and roentgenologic studies have not shown that workers in the MMVF industry have demonstrable pulmonary changes attributable to MMVF exposures. Large scale international experimental studies have demonstrated that MMVF cause neither lung cancer, mesothelioma nor lung fibrosis when inhaled by rats. Other studies have confirmed this and extended these findings also to monkeys and hamsters. Epidemiologic studies on a total of over 40,000 workers in the MMVF industry have failed to demonstrate that exposure to MMVF is associated with a significantly increased risk of death from lung cancer or nonmalignant respiratory disease. PMID- 3026165 TI - The glycemic response to meals with six different fruits in insulin-dependent diabetics using a home blood-glucose monitoring system. AB - Glycemic effect of adding 10-g carbohydrate portions of apple, banana, grapes, honeydew, orange, or strawberries to a standard meal on separate occasions was measured in 10 insulin-dependent diabetics monitored at home. The meal comprised 29% of total daily caloric intake and contained green beans, rice, turkey, and margarine (50% carbohydrate, 20% protein, and 30% fat). Blood-glucose response to meals containing grapes, honeydew, orange, or strawberries was slightly higher than meals containing apple, banana, or no fruit and the small amount of starch in apple and banana may have contributed to their lower blood-glucose response compared to the other fruits tested. Age, duration of diabetes, or insulin regimen did not correlate with subjects' responses to fruit. PMID- 3026164 TI - Determination of total body water by deuterium NMR. AB - A new deuterium nuclear magnetic resonance (2H NMR) method is described for determining total body water in humans. The method has been validated against a standard infrared absorption (IR) procedure using a tracer dose of deuterium oxide (2H2O) of approximately 10 g for each human subject. The precision and accuracy for the methods have been compared and found to be very similar. The advantages of the 2H NMR method over other presently available techniques that are based on 2H2O dilution are as follows: it is fast, accurate, needs only a small dose of 2H2O, can be done using any body fluid, and, most importantly, does not require any sample preparation. PMID- 3026166 TI - Fatty acids: inhibition. PMID- 3026167 TI - Dietary fat and fiber: relationship to caloric intake, body growth, and colon tumorigenesis. PMID- 3026168 TI - Cytomegalovirus infection initially diagnosed by skin biopsy. AB - The reported cases of cutaneous involvement by cytomegalovirus (CMV) are mostly limited to descriptions in immunocompromised patients who had other evidence of systemic CMV infection or in patients whose disseminated infection was not diagnosed until autopsy. However, three cases of CMV infection initially diagnosed from antemortem biopsy of skin lesions have been documented in the literature. The authors review in depth one of these three cases and describe two new cases of CMV infection first identified by skin biopsy. The latter two are the first detailed descriptions of this infection in the skin of burned patients. Review of the literature describing cutaneous CMV involvement reveals protean epidermal and dermal manifestations with consistent histologic findings. Recently, however, there has been documentation of histologically occult CMV in patients with disseminated CMV infection. Skin biopsy of cutaneous lesions in immunocompromised patients is important in the diagnosis of systemic CMV infection. PMID- 3026169 TI - Primary oat cell carcinoma of the esophagus. AB - A case of primary oat cell carcinoma of the esophagus is presented. Clinical, radiological, and pathological findings in our case and those reported earlier were reviewed. Primary oat cell carcinoma of the esophagus should be suspected in patients with symptoms of dysphagia, radiating pain to the back with bulky mass on the esophagogram, and diffuse metastasis. PMID- 3026170 TI - Functional analysis of the marginating pool of human polymorphonuclear leukocytes. AB - The intravascular pool of human polymorphonuclear leukocytes (PMN) is composed of one compartment which is circulating and another that is marginated to the vascular endothelium. Administration of B-adrenergic agonists leads to a rapid demargination with an increase in the circulating PMN pool. The marginating PMN has previously been stated to represent an older PMN based on a higher cytochemical alkaline phosphatase activity. With the understanding that circulating PMN are heterogeneous with respect to function and size we undertook the present study to evaluate the contribution of the marginating PMN to functional and volume-dependent heterogeneity. We found that PMN isolated 7 min after epinephrine administration, presumably enriched by marginating PMN, were not different in volume, biochemically measured alkaline phosphatase activity, stimulated superoxide anion release, degranulation, or phagocytosis. These data suggest that the circulating and marginating pools of PMN are interchangeable and that the marginating PMN are not enriched by a particular subpopulation of PMN. PMID- 3026172 TI - Genetic analysis of cystic fibrosis: linkage of DNA and classical markers in multiplex families. AB - Linkage of cystic fibrosis (CF) to DNA and classical markers was studied in 36 families of two or three generations with at least two living affected children. Among the 79 affected children, no recombinants were detected between the disease and the markers MET and pJ3.11, previously shown to be linked to CF. No linkage between the human trypsin gene family (which appears to include at least 10 members) and CF was found, although not all genes of the trypsin family have been screened yet. In one of the CF families, recombination between MET and pJ3.11 was detected in an unaffected sib. Data from our families suggest that the gene order of markers among chromosome 7q is: (7cen;p8.33)collagen(COL1A2);DOCR1 917;paraoxonase+ ++(PON);(MET-cf-J3.11);T-cell receptor beta chain (TCRB);qter. There was no evidence for (or against) either postzygotic selection or meiotic drive to explain the high frequency of CF in Caucasian populations. PMID- 3026171 TI - Linkage of cystic fibrosis to two tightly linked DNA markers: joint report from a collaborative study. AB - A collaborative study involving seven research groups provided an opportunity to investigate the linkage relationships between cystic fibrosis and two DNA marker loci, MET and pJ3.11 (D7S8), on an extended sample of 211 tested families. The maximum lod scores, recombination estimates, and confidence upper bounds (in parentheses) were 91.0 at theta = .004 (.012) for CF and MET, 71.3 at theta = .003 (.011) for CF and D7S8, and 69.3 at theta = .018 (.036) for MET and D7S8. Three-locus analyses yielded best support for the order MET-CF-D7S8, with odds against the alternate orders CF-MET-D7S8 and CF-D7S8-MET of 9:1 and 161:1, respectively. However, the number of observed recombinants was small and only one of the recombinants was jointly informative for all three markers. Significant allelic association was found between CF and both MET and D7S8. Weaker association between the latter two loci is consistent with the order MET-CF-D7S8. PMID- 3026173 TI - Linkage of cystic fibrosis locus and polymorphic DNA markers in 14 families. AB - Linkage relationships between the cystic fibrosis (CF) locus and three polymorphic DNA markers were examined in 14 families, five of which were of Hispanic origin. Tight linkage was found between the CF locus and MET (maximum lod score = 7.16 at theta = .001), and between CF and pJ3.11 (maximum lod score = 3.87 at theta = .001). We observed two recombinations between CF and collagen, yielding a maximum lod score of 0.359 at theta = .125, and one recombination in the cluster CF-MET-pJ3.11. Analysis by the seriation method indicates the order COL-pJ3.11-CF-MET. PMID- 3026174 TI - Human acid beta-glucosidase: Northern blot and S1 nuclease analysis of mRNA from HeLa cells and normal and Gaucher disease fibroblasts. AB - Human acid beta-glucosidase (beta-Glc) mRNA was evaluated by dot blot, Northern blot, and S1 nuclease analyses of extracts of HeLa cells and cultured fibroblasts from normal individuals and Gaucher disease patients. Dot blot quantitation indicated an equal concentration of specific mRNA in all sources. Northern blot analyses demonstrated the presence of three poly (A)+ mRNAs of about 5,600, 2,500, and 2,000 nucleotides in length from all cell extracts. All three mRNAs were present in normal amounts in fibroblast extracts from several subtypes and variants of Gaucher disease. The largest poly (A)+ mRNA species was thought to represent an unspliced nuclear precursor for the two smaller beta-Glc mRNAs. S1 nuclease analyses, using SP6 transcripts of beta-Glc cDNA, indicated that the 2,000 nucleotide mRNA differs from the 2,500 nucleotide form at both the 5' and 3' ends. These results are consistent with the use of an alternate 5' splice site and 3' polyadenylation signal. These results also suggest that the subtype and variants of Gaucher disease result from single base alterations that lead to the synthesis of defective beta-Glc. PMID- 3026175 TI - Inverted duplication of JH associated with chromosome 14 translocation and T-cell leukemia in ataxia-telangiectasia. AB - A specific 14q32 breakpoint is observed in a homologous chromosome 14 translocation [t(14;14)q12q32] occurring in the T-cells of about 10% of patients with ataxia-telangiectasia (AT). To investigate whether the 14q32 breakpoint in AT occurs within the immunoglobulin gene cluster as is frequently detected in B cell lymphoma, immunoglobulin clones were hybridized to Southern blots of DNA isolated from the T-cells of two AT patients with this chromosome 14 translocation. The 14q32 translocation breakpoints in these patients are apparently not within JH, S mu, C mu, S alpha-1 or -2, or C alpha-1 or -2, but one of the patients has an inverted duplication of at least 26 kilobases (kb) of the C mu region, with an associated 5' flanking deletion. The point of origin of the inverted duplication is within JH near the recombination signal for the J4 gene. This suggests that normal JH recombination mechanisms may have played a role in the development of this 14q32 chromosomal aberration. The presence of AT chromosomal breakpoints near other rearranging genes suggests a role for exaggerated recombination in the pathogenesis of chromosomal instability in AT. PMID- 3026177 TI - Acute nonlymphocytic leukemia after treatment of small cell lung cancer. AB - From 1977 to 1982, 377 patients with small cell lung cancer were treated at Vanderbilt University Medical Center. All patients received combination chemotherapy consisting of cyclophosphamide, doxorubicin, and vincristine (CAV) with or without methotrexate, etoposide, and/or hexamethylmelamine. Thoracic and/or prophylactic cranial irradiation was administered to 159 (42 percent) and 192 (51 percent) patients, respectively. Acute nonlymphocytic leukemia was observed in two patients at 22 and 81 months from the start of therapy. The relative risk of leukemia was 154 (95 percent confidence limit, 38 to 293). A Kaplan-Meier estimate of the cumulative probability of leukemia was 1.9 +/- 1.4 percent seven years after the start of treatment. The relative risk of leukemia is significantly increased in this group of patients (p less than 0.0001). Acute nonlymphocytic leukemia is a long-term complication of small cell lung cancer therapy. PMID- 3026176 TI - Genetic studies of low abundance human plasma proteins. III. Polymorphism of the C1R subcomponent of the first complement component. AB - Genetic polymorphism of the C1R subcomponent of human complement component C1 has been detected in normal plasma samples using the high resolving power of isoelectric focusing in 6 M urea followed by immunoblotting. There are two common alleles at the C1R structural locus that show autosomal codominant inheritance. The C1R*1 and C1R*2 allele frequencies in U.S. white and U.S. black blood donors are: .934, .066, and .899, .101, respectively. PMID- 3026178 TI - Viral particles in the conjunctiva of a patient with the acquired immune deficiency syndrome. AB - Conjunctival biopsy from a 39-year-old man with the acquired immune deficiency syndrome revealed the presence of herpes virus particles by electron microscopy. The finding of herpes virus particles in the conjunctiva may be of prognostic significance in documenting the presence of systemic viral infection. PMID- 3026179 TI - Peripheral neuropathy as a component of the neuroleptic malignant syndrome. AB - The neuroleptic malignant syndrome, an idiosyncratic reaction to antipsychotic medication, is defined by the tetrad of autonomic dysfunction, rigidity, fever, and altered mental status. Two patients in whom this syndrome developed acquired severe peripheral neuropathies during the course of their illness. This demyelinative polyneuropathy has not been reported previously as occurring in the neuroleptic malignant syndrome. PMID- 3026180 TI - Rebound hypertension after discontinuation of transdermal clonidine therapy. AB - Three (and possibly all four) elderly hypertensive patients who were followed sequentially after discontinuation of transdermal clonidine monotherapy manifested a rapid rise in blood pressure to levels above control (pre- and post therapy) readings. No signs of an "overshoot" in plasma norepinephrine levels or symptoms of beta-adrenergic overactivity were seen. Such rebound hypertension suggests hypersensitivity to alpha-adrenergic receptor stimulation and could pose a heretofore unreported potential hazard to elderly or otherwise compromised patients who discontinue transdermal clonidine therapy. PMID- 3026181 TI - Germ cell tumors and leukemia. PMID- 3026183 TI - Clinical high-risk designation does not predict excess fetal-maternal hemorrhage. AB - During a 5-year period, an enzyme-linked antiglobulin test was used to screen and quantitate fetal-maternal hemorrhage in 789 consecutive D-negative mothers who were delivered of D-positive babies. Six hundred seventy-two patients (85.2%) had no detectable fetal-maternal hemorrhage, and 117 patients (14.8%) had a detectable fetal-maternal hemorrhage. Eight of the 789 (1%) had a fetal-maternal hemorrhage greater than 30 ml and required more than one vial of Rh immune globulin. Two patients with fetal-maternal hemorrhage of 29 and 30 ml also received additional Rh immune globulin. Each case was reviewed for the presence of high-risk features that are thought to predict patients at risk for fetal maternal hemorrhage. Patients having a cesarean section or complicated vaginal delivery were considered to be in a group at high risk for fetal-maternal hemorrhage, while those with a spontaneous vaginal delivery were considered not to be at risk for fetal-maternal hemorrhage. Thirty-two of 237 patients (13.5%) in the risk group and 82 of 552 patients (15.3%) in the group not at risk had detectable fetal-maternal hemorrhage. The incidence of fetal-maternal hemorrhage for these two groups was not statistically different (p greater than 0.50 by chi 2 analysis). If only patients in the risk group had been screened for fetal maternal hemorrhage, then five of 10 (50%) who required more than one vial of Rh immune globulin would have been undertreated and at risk for developing anti-D antibodies. In addition, newborn birth weight, Apgar scores, and hematocrits were examined for 13 cases of fetal-maternal hemorrhage of greater than or equal to 21 ml, and none of these characteristics could be used to predict patients at risk for fetal-maternal hemorrhage. Therefore, no maternal or newborn characteristics could be identified that would reliably predict patients at risk for fetal maternal hemorrhage. We conclude that all D-negative patients with D-positive babies should continue to be screened for fetal-maternal hemorrhage to identify those patients requiring more than one vial of Rh immune globulin. PMID- 3026182 TI - Clinical significance of serum thermolysin-like metalloendopeptidase and its relationship to serum angiotensin converting enzyme in sarcoidosis. AB - Thermolysin-like metalloendopeptidase (TME) is an oligopeptidase that, like serum angiotensin converting enzyme (ACE), is elevated in patients with active sarcoidosis. It has previously been shown that TME and ACE have similar validity as markers of disease activity, and that when used concurrently, they complement each other in the detection of active disease. In the present study, serum TME and ACE were measured in 122 consecutive patients with sarcoidosis, and the relationship of these enzymes to radiographic stage, disease chronicity, and corticosteroid treatment was examined. Highest values of TME and ACE were seen in patients with hilar adenopathy and parenchymal disease (roentgenographic stage II) (p less than 0.05). The mean TME value for patients with chronic disease (at least two years' duration, with parenchymal infiltrates and/or persistent lymphadenopathy, splenomegaly, uveitis, or bone lesions) was twice that of all other patients (p less than 0.001). In contrast, ACE was not significantly increased in chronic disease. Follow-up measurements of these enzymes in patients subsequently treated with corticosteroids revealed that TME values did not significantly change with treatment, whereas ACE decreased by 50 percent (p = 0.005). In summary, TME and ACE are markers of active sarcoidosis that are most elevated in patients with radiographic stage II disease. TME differs from ACE in that TME is elevated in patients with chronic sarcoidosis and it is not affected by administration of corticosteroids. The data indicate that TME and ACE may reflect different aspects of the disease process. PMID- 3026184 TI - Labial fusion in prepubescent girls: a marker for sexual abuse? AB - Review of more than 500 medical records of patients referred for sexual abuse evaluations revealed 10 cases of labial fusion. Patients ranged in age from 2 months to 5 years, and the duration of the fusion ranged from 2 weeks to 2 1/2 years. No child had vulvovaginitis, dermatitis, or known genital trauma (for instance, straddle injury). Urinary tract problems were present in three patients. History and/or physical findings were consistent with sexual abuse in six of the 10 patients. Anal findings were grossly abnormal and consistent with anal penetration in all patients with a positive finding on examination. Therapy with conjugated estrogen cream was instituted in nine patients, with resolution of symptoms occurring in six. The exact cause of labial fusion is unclear. Fusion secondary to fecal soiling may occur in the young infant. Older girls may develop fusion after trauma such as that associated with sexual abuse, particularly vulvar coitus. PMID- 3026185 TI - Sexually transmitted papillomaviral infections. I. The anatomic distribution and pathologic grade of neoplastic lesions associated with different viral types. AB - Multiple colposcopic biopsy specimens were collected from 160 women, with sampling of principal cervical and vulvar lesions as well as secondary areas of either minor acetowhitening or normal epithelium. Papillomaviral deoxyribonucleic acid was detected by Southern blot hybridization in 197 (90%) of the 218 principal biopsy specimens and 93 (46%) of 198 secondary biopsy specimens. Although different papillomaviruses were found at different sites in 31 women, only six of 416 specimens contained multiple types within the same sample. Specific viral types were associated with specific disease patterns. Only one of 80 type 6 or 11 infections had a diagnosis greater than cervical intraepithelial neoplasia, grade 2. In contrast, 42 of 48 (90%) biopsy specimens of cervical intraepithelial neoplasia, grade 3, or invasive cancer contained type 16, 18, or 31. Nonetheless, 12 of 124 (10%) cases of condyloma and cervical intraepithelial neoplasia, grade 1, were associated with types 16, 18, and 31 infections. Of 58 women with multicentric disease, 46 had positive hybridizations for both cervical and vulvar lesions (32 showing the same type in both samples and 14 showing different viruses). Differing patterns of papillomavirus-induced disease arise partly from the predilection of specific viral types for certain anatomic sites and partly through variations in host response. Detection of viral deoxyribonucleic acid in 46% of the secondary biopsy specimens suggests that disease expression may represent focal breakdown of host surveillance within a field of latent papillomaviral infection. PMID- 3026186 TI - Use of intravitreal ganciclovir (dihydroxy propoxymethyl guanine) for cytomegalovirus retinitis in a patient with AIDS. AB - A patient with acquired immune deficiency syndrome with bilateral cytomegalovirus retinitis was treated with intravitreal 200-micrograms/0.1-ml doses of ganciclovir (9-[2-hydroxy-1-(hydroxymethyl) ethoxymethyl]guanine). The ganciclovir serum and intravitreal concentrations were measured with an enzyme linked immunosorbent assay and pharmacokinetic factors were determined. There was no evidence of systemic absorption of ganciclovir from the eye. The elimination half-life of ganciclovir from the vitreous was estimated to be 13.3 hours. The intravitreal concentration remained above the ID50 of cytomegalovirus for approximately 62 hours after a single injection. Clinically, the patient retained useful vision in his right eye for three months. A total of 28 intravitreal injections were given on an outpatient basis under topical anesthesia and were well tolerated. There was no evidence of retinal toxicity from the drug. PMID- 3026187 TI - Viability of herpes simplex virus type 1 on the applanation tonometer. AB - We performed several experiments to define possible factors that influence the viability of herpes simplex virus type 1 on the applanation tonometer. Infectious virus could be detected on experimentally inoculated tonometer heads for up to two hours during natural drying. If tonometers were kept moist the virus could be detected more than eight hours later. Several ophthalmic solutions, including topical anesthetics, dilating agents, and a fluorescein solution, showed only minimal antiviral activities. Wiping of virus-infected tonometer heads with a dry tissue was ineffective and allowed residual infectious virus to remain. However, no infectious virus could be detected on infected tonometer heads that had been swabbed with 70% isopropyl alcohol. Our results raised concerns regarding effective disinfection of tonometers after eye examinations. We concluded that the applanation tonometer should be swabbed routinely with alcohol after each patient examination to ensure complete virus inactivation and to prevent accidental transmission of virus in the clinical setting. PMID- 3026189 TI - Stocks of variola virus. PMID- 3026188 TI - Gossypol-induced damage to mitochondria of transformed Sertoli cells. AB - Studies on gossypol-induced morphologic changes in transformed Sertoli cells (TM4) were performed at both light- and electron-microscopic levels. Exposure of TM4 cells to 5 micrograms gossypol/ml for greater than 1 hour has severe, deleterious effects on the structure and function of mitochondria. Mitochondrial function in TM4 cells was monitored by employing a fluorochrome, Rhodamine 123, which accumulates rapidly in mitochondria having a high transmembrane potential. In gossypol-treated TM4 cells, Rhodamine 123 mitochondrial staining was reduced significantly 1 hour after the drug addition and reached a minimal level at 3 hour. Concomitantly, cytoplasmic vacuoles were detected even at the light microscopic level. Electron-microscopic studies revealed that these vacuoles were distended mitochondria. The morphology of these damaged organelles changed gradually, starting with the transformation of the tubular mitochondria into the rounded forms. Cristae concurrently collapsed onto the organelles' periphery. In addition, the ground matrixes disappeared, and the mitochondria appeared as empty vacuoles. Further evidence that these vacuoles were distended mitochondria was derived from the cytochemical localization of cytochrome c oxidase in these vacuole-like structures. PMID- 3026190 TI - Polygraphical study on age dependent epileptic encephalopathy--relationship between body movements during sleep and prognosis. AB - Body movements (BMs) during sleep in patients with age dependent epileptic encephalopathy (ADEE) were studied polysomnographically in order to clarify the underlying mechanism of intractability and the age dependent trend. Twenty patients were divided into two groups according to the prognosis of convulsions. In the good prognosis group, BMs were nearly normal except for a low frequency in some cases. In the intractable group with seizures which were uncontrollable by medication and recurred within a year, BMs showed abnormalities as follows: abnormal distribution according to sleep stages, and/or a low frequency; increased BMs on therapy with prednisolone or ACTH. Moreover, a paradoxical increase of BMs with age and recurrence of seizures concomitantly occurred in the course of the disease. Status epilepticus appeared in cases under 1-DOPA administration or with a strikingly high frequency of BMs. Since electrophysiological evidence indicates that BMs during sleep are modulated by the dopaminergic (DA) system, the present data might suggest that prognosis of convulsions in ADEE depend upon, at least in part, the DA system. And denervated supersensitivity of that system might give rise to recurrence of seizures and status epilepticus. PMID- 3026191 TI - Suppression of 2', 3'-cyclic nucleotide 3'-phosphohydrolase activity in suckling rat brain by ACTH. AB - Administration of ACTH at two different doses (0.05 and 0.5 microgram/g/day) to suckling rats resulted in the suppression of both the body and brain weight gains and the developmental increase in brain CNPase activity, and the suppression of the brain CNPase activity persisted for 3 weeks (up to the end of the experiment) after the cessation of ACTH administration in the suckling period, while the suppression of the body and brain weight gains was noticed only during the administration period. The authors emphasized the possibility that long-term therapy with massive doses of ACTH for infantile spasms may be hazardous to the developing brain in many ways. PMID- 3026192 TI - Acute lymphoblastic leukemia in Argentina. Relationship between surface markers and prognosis. AB - Leukemic blasts from patients with acute lymphoblastic leukemia (ALL) were tested initially for the following surface markers: sheep erythrocyte receptors at 4 degrees C and 37 degrees C; receptor for the C3 fraction of complement; mouse erythrocyte receptor; and surface immunoglobulins. We found that 73% of the ALL were devoid of these markers, 9% had C3 receptor only, 2% were B-ALL, and 15% were T-ALL. Only one of 147 cases tested expressed receptors for mouse erythrocyte receptor. C3-ALL predominated in girls and the prognosis was similar to that of the ALL devoid of markers. T-ALL was associated with some high-risk factors, such as high initial leukocyte counts, and a predominance in boys older than 5 years of age. Further studies were done in which cytoplasmic immunoglobulins were evaluated and surface antigens were identified by the following monoclonal antibodies: OKT1, OKT3, OKT4, OKT6, OKT8, OKT9, OKT10, OKT11a, OKIa1, and J5 (anti-CALLA). Less than 1% were B-All, 15% were T-ALL, and approximately 85% were non-T, non-B ALL. Of the latter group, 69% were common ALL, 16% were pre-B ALL, and 15% were null-ALL. Within the T-ALL, the expression of the T antigenic mosaic was heterogeneous. Results by a multivariate analysis demonstrated that sex, age, initial leukocyte count, and T-cell phenotype were independent variables with a negative prognostic value (p less than 0.01), and the only combination with significant interaction was between T-cell phenotype and leukocyte counts. PMID- 3026193 TI - Hepatocellular carcinoma in an 11-year-old girl with Fanconi's anemia. Report of a case and review of the literature. AB - An 11-year-old girl with Fanconi's anemia, who died of Corynebacterium septicemia, was found at autopsy to have a solitary, previously undiagnosed hepatocellular carcinoma (HCC). Although the association between Fanconi's anemia and malignancies such as leukemia and squamous cell carcinoma is well documented, its relationship to HCC remains controversial and obscure. Anabolic steroid therapy for Fanconi's anemia has also been considered a promoter for hepatocellular neoplasms. This report documents the youngest known patient with Fanconi's anemia to develop HCC and discusses the association between these conditions. PMID- 3026194 TI - Bronchoscopic diagnosis of cytomegalovirus pneumonia following pediatric bone marrow transplantation. AB - A pediatric bone marrow transplant (BMT) patient with cytomegalovirus (CMV) pneumonia is reported. The diagnosis was made with flexible fiberoptic bronchoscopy and bronchoalveolar lavage (BAL). The use of BAL in diagnosing CMV pneumonitis in an immunocompromised child after BMT is discussed. PMID- 3026195 TI - Mammary and extramammary Paget's disease. AB - Ten cases of mammary Paget's disease and 10 cases of extramammary Paget's disease were studied for differences of histologic features in them. Based on the epidermal changes alone, Paget cells in 90% of specimens from lesions of extramammary Paget's disease had abundant mucin that stained well, whereas Paget cells in only 40% of specimens from mammary Paget's disease stained for mucin and faintly at that. In 60% they did not stain at all. The conclusion derived from these observations is that the mucin in Paget cells of extramammary Paget's disease is different from that found in cells of mammary Paget's disease. The Paget cells in these two conditions seem to have different origins, i.e., those of mammary Paget's disease ascend to the epidermis from lactiferous ducts, whereas those of extramammary Paget's disease originate in the epidermis itself. PMID- 3026196 TI - Mammary Paget's disease without underlying carcinoma. PMID- 3026197 TI - Reduced sensitivity of red blood cell (Na+,K+)-ATPase to ethanol in vitro in male alcoholic patients: relationship to clinical characteristics. AB - We examined red blood cell (Na+,K+)-ATPase, its sensitivity to inhibition by ethanol in vitro, and its relationship to clinical characteristics and history in 41 newly admitted alcoholic patients and 14 age-matched healthy controls. Sensitivity to ethanol was significantly lower in the alcoholic patients and correlated negatively with ethanol intake. In addition, sensitivity of enzyme to ethanol was lower in patients with high agitation-anxiety ratings and correlated negatively with agitation and anxiety scores on the Brief Psychiatric Rating Scale. There were no relationships between (Na+,K+)-ATPase measures and depressive symptoms, history of treatment for depression, or family history of depression. These data suggest that tolerance to the effects of ethanol on (Na+,K+)-ATPase occurs in man and may be related to the severity of ethanol dependence or withdrawal. PMID- 3026198 TI - The feeding of ethanol in liquid diets. PMID- 3026199 TI - [Lithium and catecholaminergic neurotransmission]. PMID- 3026200 TI - Artifactual phosphate binding due to impurities in [32P]orthophosphate. AB - Many commercial preparations of [32P]orthophosphate contain radioactive impurities that interfere with binding and transport studies in biological systems. One type of impurity is micro-particulate whereas another may be pyrophosphate. Methods of removing these impurities from radiolabeled orthophosphate solutions are described. PMID- 3026201 TI - High-performance liquid chromatographic determination of 6-aminopenicillanic acid by postcolumn alkaline degradation. AB - A high-performance liquid chromatographic method has been developed for the determination of 6-aminopenicillanic acid in amino acid mixtures and human serum. The separation of 6-aminopenicillanic acid was carried out on a C18 column using sodium heptylsulfonate or tetrabutylammonium bromide as an ion-pairing agent and methanol as an organic mobile phase modifier. Detection was based on a postcolumn reaction with sodium hydroxide, mercury(II) chloride, and ethylenediaminetetraacetic acid disodium salt followed by measurement of ultraviolet absorbance (at 300 nm) of the reaction product(s). The method is quantitative for 6-aminopenicillanic acid concentrations down to 0.1 microgram/ml in human serum samples with a 20-microliter injection. At a concentration of 2 micrograms/ml, the within- and between-run precisions (relative standard deviation) were 1.29-3.91% and 2.30%, respectively. PMID- 3026202 TI - A simple procedure for the preparation of pure kinetoplast DNA network free of nuclear DNA from the kinetoplast hemoflagellate Leishmania donovani. AB - A simple, inexpensive procedure for preparing pure kinetoplast DNA network from Leishmania donovani is described. L. donovani promastigotes were lysed by incubating with pronase in presence of sodium dodecylsulfate. Crude kinetoplast DNA networks were obtained by centrifugation of the lysate through a 20% sucrose solution. The pellet containing kinetoplast DNA was deproteinized by phenol extraction. Contaminating nuclear DNAs were removed by denaturation with alkali, neutralization, and addition of polyethylene glycol-8000 to a concentration of 10% to facilitate precipitation of kinetoplast DNA. kDNA isolated after centrifugation was deproteinized several times with phenol and finally precipitated with ethanol. The average yield by this procedure is 30-50 micrograms of kDNA per gram of wet cells. By slot-blot hybridization with a nuclear DNA probe, no nuclear DNA contamination of the kDNA networks could be detected. PMID- 3026203 TI - Protein crosslinking reagents containing a selenoethylene linker are cleaved by mild oxidation. AB - A homobifunctional cleavable crosslinking reagent containing a selenoethylene group in the linker, and related reagents, have been synthesized and tested in a model system involving formation of a complex between albumin and cytochrome c. Functionally, complex formation was suggested by albumin inhibition of the ascorbate reduction of cytochrome c. Structurally, complex formation was demonstrated by crosslinking and subsequent separation of crosslinked complex from non-crosslinked proteins by SDS-polyacrylamide gel electrophoresis. The crosslinks were found to be cleavable by mild oxidation with low concentrations of periodate or with N-chlorobenzenesulfonamide immobilized on polystyrene beads (Iodo-Beads). PMID- 3026204 TI - Cytodifferentiation and immunocharacteristics of adenohypophysial cells in the toad, Bufo melanostictus. AB - Employing the unlabelled antibody enzyme technique cytodifferentiation, immunocharacteristics and topographical distribution of melanotropic (MSH), adrenocorticotropic (ACTH), thyrotropic (TSH), prolactin (PRL), gonadotropic (GTH) and growth hormone (GH) secreting cells in the embryonic/larval as well as adult pituitary gland of the common Indian toad, Bufo melanostictus, have been studied by using antisera raised in rabbit against mammalian hypophysial hormones. Immunoreactive MSH and ACTH cells appear first in the dorsocaudal and rostral regions of the pituitary anlage (PA) at stage 21 (Gosner's classification) of the embryonic development. This is followed by the differentiation of TSH and PRL cells at stage 22 in the midventral and central regions of the PA respectively. Finally, at stage 23 the GTH cells appear in the rostral and the GH cells in the caudal regions of the PA. With the progress of the development, cells showing immunoreactivity to various antisera gradually increase in number, size, granular content and finally occupy the characteristic adult disposition. The MSH cells comprise the pars intermedia. In the pars distalis (PD) the ACTH cells are localized in the rostroventral region, TSH cells in the central region and the GH cells in the dorsocaudal region. However, GTH and PRL cells are distributed throughout the PD. PMID- 3026205 TI - Projection from the inferior colliculus to the superior olivary complex in the albino rat. AB - The origin and termination of the fibers projecting from the inferior colliculus to the superior olivary complex have been studied in rat by means of the Fink and Heimer and horseradish peroxidase techniques (anterograde and retrograde transport of free horseradish peroxidase and peroxidase conjugated to wheat germ agglutinin). The colliculoolivary fibers originate in layer 3 of the external cortex and the adjacent part of the central nucleus, particularly in the former. Via the lateral lemniscus the fibers reach the ipsilateral periolivary region where they terminate in the rostral and medioventral zones. The terminal field contains at least two types of cells which could constitute the next link in the descending projection to the cochlea and/or the cochlear nuclei. One of these is the large olivocochlear neuron, and the other a smaller neuron projecting to the cochlear nuclei. Judged by their topographic relationship in the inferior colliculus and in the superior olive, the colliculoolivary neurons may form a link in a oligosynaptic projection from the auditory cortex to the cochlea and/or the cochlear nuclei. The observations are based on light microscopy, however, and do not allow conclusions concerning synaptic contacts. PMID- 3026207 TI - Absence of beta-adrenergic receptor involvement in cerebrovascular dilation by halothane in monkeys. AB - We determined, in monkeys, whether halothane-induced cerebrovascular dilation is mediated by beta-adrenergic receptors and whether cerebrovascular tone progressively returns to baseline values during prolonged halothane anesthesia. Total cerebral blood flow (CBF), cerebral perfusion pressure, plasma halothane concentration, and arterial blood gas tensions and pH were measured in 14 rhesus monkeys mechanically ventilated with 0.5% (inspired) halothane, 33% O2 and balance N2O. Halothane was increased to 2.0% and the measurements repeated 30 and 60 min later. Then either 0.9% NaCl (controls n = 6) or propranolol (n = 8), 1.0 mg/kg was infused intravenously over 10 min, and the measurements repeated at 70, 90, 120, and 150 min. After 30 min at 2.0% halothane, CBF increased in the controls by 50% (P less than 0.05) from 92 +/- 8 (mean +/- SD) to 137 +/- 39 ml X 100 g-1 X min-1 and in the propranolol group by 30% (P less than 0.05) from 106 +/- 33 to 137 +/- 28 ml X 100 g-1 X min-1. After 2.5 hr of 2.0% halothane anesthesia, CBF remained elevated above baseline levels, but by only 28 and 23% in the control and propranolol groups, respectively. Cerebrovascular resistance was identical in both groups (0.55 +/- 0.33 vs 0.53 +/- 0.13 mm Hg X ml-1 X 100 g 1 X min 1). The results show that there is only a 10-20% return of CBF toward baseline levels after up to 2.5 hr of 2% halothane anesthesia. The results also indicate that halothane-induced cerebrovascular dilation is not mediated by beta adrenergic receptors. PMID- 3026209 TI - Acquired immunodeficiency syndrome: challenge to the infection control practitioner. PMID- 3026208 TI - Neuromuscular blockade of atracurium versus succinylcholine in a patient with complete absence of plasma cholinesterase activity. PMID- 3026206 TI - Thalamic projections to the posterior sylvian and posterior ectosylvian gyri of the sheep brain, revealed with the retrograde transport of horseradish peroxidase. AB - The auditory area of the sheep cerebral cortex was studied on the basis of its afferents from the medial geniculate nucleus, traced with the horseradish peroxidase retrograde transport method. The results show that the medial geniculate nucleus projects only to the anterior parts of the posterior ectosylvian gyrus and the posterior sylvian gyrus. A small area of the posterior ectosylvian gyrus receives afferents exclusively from the ventral part of the medial geniculate nucleus, while the anterior part of the posterior sylvian gyrus receives also afferents from the posterior nucleus of the thalamus and the pulvinar. In addition, it was found that the medial part of the medial geniculate nucleus projects in a sparse way to the auditory cortex. The middle part of the posterior ectosylvian gyrus receives afferents from the posterior nucleus of the thalamus, the suprageniculate nucleus and the pulvinar, while the posterior part of the posterior ectosylvian gyrus together with the posteriormost part of the posterior sylvian gyrus receive afferents from the pulvinar. Finally, the area located between the anterior and the posteriormost part of the posterior sylvian gyrus receives afferents from both the posterior nucleus of the thalamus and the pulvinar. PMID- 3026210 TI - The mechanism of corticosteroids in treating asthma. AB - It would be difficult for physicians or allergists to imagine doing without corticosteroids in managing difficult cases of bronchial asthma. It is beyond any doubt that CS act on many sites to help reverse the pathologic process of bronchial asthma. Corticosteroids enhance the beta-adrenergic response to relieve the muscle spasm. They also act by reversing the mucosal edema, decreasing vascular permeability by vasoconstriction, and inhibiting the release of LTC4 and LTD4. Corticosteroids reduce the mucus secretion by inhibiting the release of secretagogue from macrophages. Corticosteroids inhibit the late phase reaction by inhibiting the inflammatory response and interfering with chemotaxis. This action may be due to the inhibition of LTB4 release. The eosinopenic effect of corticosteroids may help to prevent the cytotoxic effect of the major basic protein and other inflammatory mediators released from eosinophils. Corticosteroids have no effect on the immediate hypersensitivity reaction and have no direct role in bronchial reactivity. By blocking the late reaction, they prevent the increased airway reactivity observed with late bronchial reactions. The limitation of using corticosteroids are their side effects. They vary from tolerable to life threatening side effects. Each tissue in the body is a target for corticosteroids. The mechanism of adverse effects have been studied in extensive detail but many questions are yet to be answered. Alternate-day therapy and inhalation therapy are meant to minimize these side effects. The expansion of using inhaled steroid therapy and finding some inhaled preparations that have even less systemic side effects seems a reasonable approach to deal with severe asthma. PMID- 3026212 TI - Restriction endonuclease patterns of bovine herpesvirus type 1 isolated from bovine mammary glands. AB - Restriction endonuclease analysis was used, in conjunction with viral neutralization and growth-curve experiments, to compare a bovine herpesvirus type 1 (BHV-1) isolate, originally obtained from bovine mammary gland lesions, with a standard BHV-1 strain, infectious bovine rhinotracheitis virus. Although differences were not detected by viral neutralization or growth-curve experiments, restriction fragment patterns generated by Bam HI, Eco RI, Hind III, and Hpa I, revealed definite differences between the isolate and the prototype strain. Additionally, Eco RI, Hind III, and Hpa I patterns revealed that the mammary gland isolate had DNA-fragment patterns characteristic of infectious pustular vulvovaginitis strain of BHV-1, type 2b. Seemingly, type-2b isolates, similar to types 1 and 2a, may be capable of causing divergent types of infection of variable severity in cattle. PMID- 3026211 TI - Ketoconazole-induced changes in selected canine hormone concentrations. AB - The effects of administering ketoconazole at a high dosage (30 mg/kg of body weight/day) and at a low dosage (10 mg/kg/day) on steroidogenesis in the dog were compared. Ketoconazole significantly suppressed basal plasma cortisol concentrations (P = 0.001), cortisol responsiveness to ACTH (P = 0.002 to 0.005), and serum testosterone concentrations (P = 0.0005). The data indicated a rebound effect after ketoconazole treatment was stopped and that testosterone suppression was being overridden at lower ketoconazole doses. Plasma 17-alpha hydroxyprogesterone concentrations (P = 0.0005) and serum progesterone concentrations (P = 0.014 to 0.003) were significantly increased during ketoconazole treatment. Aldosterone, 11-desoxycortisol, and 17-beta-estradiol concentrations did not change significantly during ketoconazole treatment. PMID- 3026213 TI - Hepatitis A virus in stool during clinical relapse. AB - Among 256 patients with acute hepatitis A, 17 (6.6%) had a relapse of the disease between 30 and 90 days after the primary episode. We studied 7 of these patients. Serologic testing showed mean alanine aminotransferase levels of 1668 IU/L during the acute stage, 107 IU/L during the early convalescence, and 1027 IU/L during the relapse. Tests for IgM antibody against hepatitis A virus were positive in the 7 patients at the onset of disease, with decreasing levels in 3 of the 4 patients tested during the evolution of the illness. Stools collected during the relapse phase showed hepatitis A virus by immune electron microscopy, radioimmunoassay, and molecular hybridization using a 32P-labeled cDNA-hepatitis A virus probe. Stools collected from 4 of these patients 6 to 12 months after the onset of disease were negative for the virus. The finding of hepatitis A virus in the stool of these patients during the relapse phase strongly implicates hepatitis A virus as the causative agent of the clinical relapse. PMID- 3026214 TI - Public health applications of a classification system for human immunodeficiency virus infection. PMID- 3026216 TI - The public health perspective on alcoholism. PMID- 3026215 TI - Enalapril in pheochromocytoma. PMID- 3026217 TI - Ethnic/religious differences in the manifestation and treatment of alcoholism. PMID- 3026218 TI - Release of endogenous amino acids, dopamine, and cyclic AMP from the rat brain: methodological aspects and mutual interferences. AB - The release from the rat brain of endogenous dopamine, its metabolites, cyclic AMP, transmitter, and other amino acids was studied with push-pull cannula perfusion or dialysis. The amino acid output from the striatum with either perfusion technique was virtually identical. Relative to the tissue content, the egress of the dopamine metabolites and of the non-transmitter (metabolic) amino acids was higher than that of the transmitter substances. The intracellular second messenger cyclic AMP was also released into the perfusate, and its content was enhanced by local application of various biogenic amines. During and after electroconvulsive shocks the release (and presumably the formation) of alanine was enhanced, which may reflect increased glycolysis and transamination in the brain. Following local application of tricyclic antidepressants, the content of transmitter amino acids in push-pull perfusates of the rat thalamus and striatum was enhanced several fold. Such an increase was not found following systemic injection of high doses of mianserin and desmethylimipramine. In contrast, the effect of amphetamine on dopamine release was seen after both local and systemic applications. Locally applied dopamine produced a specific decrease in the release of gamma-aminobutyric acid in both the substantia nigra and in the striatum. Muscimol decreased, whereas picrotoxine enhanced the release of GABA from the rat striatum. Our observations emphasize that transmitter release and brain metabolism can be monitored by perfusion techniques and that the effects of drugs may depend on the route of administration. As has frequently been shown with in vitro techniques, our data indicate the importance of local control of transmitter release in vivo by both autoreceptors and alloreceptors in addition to firing activity. PMID- 3026219 TI - Embryonic signals in pregnancy. PMID- 3026220 TI - Regulation of lactate dehydrogenase gene expression by cAMP-dependent protein kinase subunits. AB - The studies described in this report suggest a rather complex, albeit incomplete, sequence of molecular events that we believe form part of the cascade of reactions through which a series of hormones, via cAMP, regulates the expression of specific gene products. The majority of our own studies relate to cAMP mediated induction of LDH. Some, if not all, of the molecular steps discussed in this paper may ultimately be recognized as part of a universal mechanism by which cAMP controls gene expression in higher eukaryotes. The idea of a functional role for cAMP-dependent protein kinase subunits in cAMP-mediated gene control has already had experimental support, but our identification of the regulatory subunit RII as a topoisomerase now more firmly points to a complex function for the kinase in regulating gene function at the DNA level. We look forward to the elucidation of the function of those nuclear proteins that serve as substrate for the catalytic subunit of cAMP-dependent protein kinase. Further studies related to the molecular interaction of RII with chromosomal DNA should be a fruitful area for future research. PMID- 3026221 TI - Molecular genetic analysis of cAMP-dependent protein kinase. AB - Genetic evidence has been obtained indicating that the responsiveness of CHO cells to cAMP requires an intact cADepPK system. DNA carrying mutant RI and C subunit genes has been transferred to wild-type cells. Recipient cells carrying the DNA have been detected by selecting for expression of the phenotype for cAMP resistance. We are in the process of cloning a functional mutant RI subunit to use as a moveable genetic element that can confer cAMP resistance on recipient cells. This cloned gene should allow us to test hypotheses concerning the mechanism of cAMP regulation of transcription in mammalian cells. PMID- 3026222 TI - Multihormonal regulation of phosphoenolpyruvate carboxykinase gene transcription. The dominant role of insulin. PMID- 3026223 TI - The relationship between structure and function in cAMP-dependent protein kinases. PMID- 3026225 TI - [Critical study of an assay of human leukocyte and lymphoblastoid interferon on MDBK cells, in microplaque, by the "dye uptake" method. I. Biological factors]. PMID- 3026224 TI - Differential expression of the genes for the mitochondrial and cytosolic forms of phosphoenolpyruvate carboxykinase. PMID- 3026226 TI - [Free radicals and myocardial ischemia]. PMID- 3026227 TI - [Metabolic disorders generated by myocardial ischemia: implication of oxygen free radicals]. AB - A process of myocardial ischemia is developed when there is an imbalance between the tissues needs and the supply in coronary arterial blood. This imbalance causes rapidly a cascade of biochemical and metabolic events which, when the ischemia is particularly severe and extended, may lead, at the end, to tissue necrosis. However, the factors responsible for cellular death are still not precisely known and it seems, from the recent experimental results, that the exacerbated production of free radicals derived from oxygen could be one of the elements implicated in the initiation of the necrotic process. This article proposes to describe the various disturbances of the myocardial metabolism induced by the ischemic process and to consider the eventual role of the oxygen free radicals in the development of these alterations. PMID- 3026228 TI - [Perspectives of in-situ measuring of oxygen free radicals]. AB - It has been proposed that the oxygen free radicals (superoxide and hydroxyl radicals), could be partially responsible for the appearance of cellular alterations in the course of various pathological situations. It has not been possible to measure these kinds of radicals because of their very short life span and the very low concentrations present in the tissues. The paramagnetic electronic resonance (PER), of which the principle lies on the specific detection of free radicals, proves, however, to be ill suited for the measurement of the oxygen free radicals, because of an insufficient sensitivity. This sensitivity may be increased with the "spin trapping" technique, the principle of which is to use a trapping component able to be associated to the unstable radical to form a stable product, which accumulates then in the tissues up to a concentration higher than the detection threshold in PER. PMID- 3026230 TI - [The pharmacology of various classes of cerebral opioid receptors]. AB - The research on endogenous opioid is only a decade old but the considerable number and the variety of studies devoted to this subject suggest that these neuropeptides might play a pivotal role in various biological functions. The most abundant opioid peptides enkephalins are synthesized as large precursors. They bind to several classes of receptors as mu and delta types and are degraded by specific enzymes (aminopeptidase M, enkephalinase, dipeptidylaminopeptidase) belonging to the group of metallopeptidases. The analysis of the functions of the enkephalinergic system can now be investigated by using recently designed selective mu (DAGO, TRIMU 5), delta (DTLET, DEPDPE), kappa (U 50, 488) agonists or antagonists (ICI 174, 864 for the delta type) and kelatorphan a complete inhibitor of enkephalin metabolism. The former probes were obtained by a rational approach based on the conformational adaptability of the endogenous peptides while inhibitors of enkephalin degrading enzymes were designed by taking into account crystallographic data on metallopeptidases. mu and delta receptors present distinct distributions in the brain. Enkephalinase visualized by autoradiography seems to be closely associated with opioid receptors. Pain control could be insured in brain structures by mu receptor-stimulation whereas both mu and delta types might be involved at the level of the spinal cord. In both cases, a "physiological" analgesia is produced by kelatorphan. PMID- 3026229 TI - [Adrenal reactivity to prolonged-action ACTH in hirsute women treated with 50 mg of cyproterone acetate combined with percutaneous estradiol]. AB - A prolonged-action Synacthen (R) test (1 mg.i.m. was performed in 14 hirsute women before and 6 months after instituting combined treatment with cyproterone acetate (50 mg/day/21 days) and percutaneous estradiol (3 mg during the last 10 days of cyproterone acetate administration). Determination of blood cortisol and precursors of cortisol--plasma androgens--was carried out to evaluate possible modification of adrenal enzymatic activity. At the used doses, cyproterone acetate failed to influence adrenal reactivity to synthetic ACTH or the enzymatic activities of 21-hydroxylase and 17-beta-reductase in vivo. PMID- 3026231 TI - Steroid hormone receptors in male breast diseases. AB - Estrogen (ER), progesterone (PR), glucocorticoid (GR) and androgen (AR) receptors were assayed in tumor samples from 8 cases of male breast cancer (MBC) and 20 cases of male gynecomastia. Seven out of eight (87.5%) male tumor samples had positive ER assays with values ranging from 12 to 180 fmol/mg protein. Of the seven ER positive cases of MBC, six, had positive PR activity with high titers. Positive GR and AR values were also detected in 75% of MBC cases. Concentrations of all four receptors were significantly correlated with each other. With gynecomastic tissue, the proportion of receptor-positive patients was 20% ER, 20% PR, 20% AR, and 45% GR. Except for GR, steroid receptor values for MBC individuals were significantly higher than those of gynecomastia patients. PMID- 3026232 TI - Modulation of membrane proteins by vertical phase separation and membrane lipid fluidity. Basis for a new approach to tumor immunotherapy. AB - Cell differentiation and proliferation entail a series of membranal events, which lead to the modulation of proteins at the cell surface. In the case of malignant differentiation this offers the tumor cell the possibility of escaping immune surveillance. Vertical phase separation of membrane proteins appears to play an important role during modulation of membrane proteins. The data reviewed here strongly suggests that the membrane lipid fluidity modulates expression of membrane proteins by vertical phase separation. When the membrane fluidity was elevated the surface expression of some membrane proteins increased, whereas it decreased when the membrane became more rigid. These proteins (e.g. H-2 antigens, hormone receptors and others were termed "syndromic". The membrane proteins which displayed the opposite behaviour with respect to the lipid fluidity were referred to as "antidromic" proteins e.g. human blood group antigens, Thy 1.2 and neuroreceptors). The possibility that the tumor cell plasma membrane contains cryptic antidromic antigens which may become exposed when the membrane lipid fluidity is manipulated has triggered a potentially new experimental approach in the treatment of neoplastic diseases. Autologous tumor cells pretreated to decrease their membrane lipid fluidity were shown to have an increased capability of eliciting specific immune responses when compared to normal control cells subjected to the same treatment. PMID- 3026234 TI - Correlation of aromatase activity and steroid receptors in human ovarian carcinoma. AB - Androgen receptors (AR) predominated (98%) in the cytosols of human ovarian carcinomas (n = 40), compared to estrogen (ER)-45%, and progestin (PgR)-53% receptors. The average age of the patients was 58 years. Serum levels of estradiol-17 beta (E2), testosterone (T), follicle stimulating hormone (FSH) and luteinizing hormone (LH) were within the normal range. One third of the cases showed a measurable aromatase activity, while in the majority of the cases this activity was undetectable. The distribution of PgR in the 40 tumors studied appeared to correlate with aromatase activity. The distribution of ER and AR did not differ significantly in aromatase positive and aromatase negative tumors. Interestingly the majority of aromatase positive cases were the endometrioid type of carcinomas. Patients with this type of ovarian carcinoma may probably benefit from anti-estrogen, progestin or aromatase inhibitor treatment. All tumors which were negative for aromatase activity contained AR. In these aromatase negative tumors androgens may not be further metabolized into estrogens and the tumor cells may be influenced by the androgens through the AR present in them. Anti androgen treatment could possibly benefit the patients with such tumors. PMID- 3026233 TI - Superoxide anion production by adriamycinol from cardiac sarcosomes and by mitochondrial NADH dehydrogenase. AB - This study investigates the effects of both adriamycin and its 13-hydroxylated metabolite adriamycinol on superoxide anion production from cardiac sarcosomes and by mitochondrial NADH dehydrogenase. Superoxide anion production was determined by using the succinoylated cytochrome c reduction assay. Both adriamycin and adriamycinol stimulated superoxide formation in cardiac sarcosomes and by mitochondrial NADH dehydrogenase. In the first case only NADPH was required as a co-factor and in the second case only NADH. From sarcosomes as well as by NADH dehydrogenase, the superoxide production followed Michaelis-Menten kinetics. With both activating enzymatic systems, the Vmax of adriamycinol was found to be similar to that of adriamycin, but the Km for the former anthracycline was higher than for the latter. Adriamycinol also increased the rate of NADPH and NADH consumption, by sarcosomal fractions and by NADH dehydrogenase respectively. At equimolar consentrations, adriamycinol consumed less NADPH and NADH than adriamycin. These results suggest that adriamycinol could contribute to the chronic cardiac toxicity of adriamycin by forming superoxide anions in cardiac cells constituents. PMID- 3026236 TI - Relevance of DNA-fluorimetry and short-term resistance testing for adjuvant treatment of non-small cell lung carcinomas. AB - In a clinical study 127 patients with previously untreated stage III non-small cell lung carcinomas (NSCLC) were investigated using flow cytometry and an in vitro short-term test for predicting resistance to cytostatic agents. Patients with aneuploid tumors and tumors with high proliferative activity had significantly shorter survival times than those with diploid or low proliferating tumors. The aim of this study was to find out whether groups of patients classified according to the additionally observed prognostic factors, experience an advantage or disadvantage from particular modalities of treatment. Seventy nine patients had surgery alone, 18 patients were treated additionally with chemotherapy, and 30 patients with radiation. Patients with aneuploid, low proliferating and in vitro resistant tumors showed no different survival rates after treatment with chemo- and radiotherapy adjuvant to surgery. In contrast, patients with high proliferating tumors died earlier under adjuvant chemotherapy and radiation. Patients with in vitro chemosensitive tumors had shorter survival times after irradiation than patients who had surgery alone or who were treated with adjuvant chemotherapy. PMID- 3026235 TI - In vitro antitumor activity of teniposide against carcinoma of the lung in human tumor clonogenic assay. AB - The antitumor activity of teniposide (VM26) on carcinoma of the lung was tested by comparing it with its congener, etoposide (VP16) in an in vitro soft agar clonogenic assay system (HTCA). Of 56 tumor biopsies placed in culture, 40 tumors were evaluable for drug sensitivity information (i.e., greater than or equal to 30 colonies in the control plate). The overall in vitro response rate (defined as less than or equal to 50% survival of TCFU) at the standard dose (10% peak plasma concentration) was 35%, which is higher than that of VP16 (28%). In cell lines derived from human carcinoma of the lung, VM26 showed 50% inhibition against colony formation of PC-7 cells at 0.45 microgram/ml which is less than that of VP16. A superior antitumor activity of VM26 on PC-9 cells was also observed. VM26 was observed to act faster than VP16 thus indicating its possible superior antitumor activity in vivo. PMID- 3026237 TI - DNA polymerases and DNA topoisomerases as targets for the development of anticancer drugs. AB - Studies of a variety of compounds designed as derivatives of prototype active molecules aphidicolin and doxorubicin are reported. So far none of the aphidicolin simpler analogues is as active as the parental molecule. Ten anthracycline analogues, characterized for their cytotoxicity, antitumor activity and inhibition of the relaxing activity of purified human DNA topoisomerase II can be divided into five groups. The majority of the tested compounds shows properties very similar to those of doxorubicin. Epirubicin shows extremely high inhibitory activity toward the relaxing property of topoisomerase II but its antitumor activity and cytotoxicity are similar to those of the former group. The third group includes a compound with extremely high cytotoxicity. The fourth group is represented by a compound which shows a cytotoxicity. The fourth group is represented by a compound which shows a cytotoxicity. The fourth group is represented by a compound which shows a cytotoxicity typical of anthracyclines and good antitumor activity but which has no specific inhibitory activity on topoisomerase II. A fifth group includes a totally inactive compound. Our results suggest that the inhibition of human DNA topoisomerase II is only partially correlated with antitumor activity. PMID- 3026238 TI - Ambulatory combination chemotherapy with oral etoposide and cisplatin for advanced non small cell lung carcinoma patients. A phase II study. AB - Thirty nine male patients with locally advanced and/or extensive non small cell lung cancer (NSCLC) were treated with oral etoposide (240 mg/m2 days 1 to 3) and cisplatin (100 mg/m2 day 4) according to a fully ambulatory schedule. Eight out of 33 (24%) evaluable patients achieved a partial response (PR) and 6 a minor response (MR). Stable disease (SD) was observed in 7 (25%) and progressive disease (P.D.) in 12 (26%). Median survival time (MST) of all patients was 8 months. No difference in MST was observed between limited (LD) and extensive disease (ED) patients. Only overall responding patients (PR + MR + SD) in the E.D. subgroup lived significantly longer than PD patients. Patients with LD did not obtain a significant survival benefit whether or not a response was achieved. The overall toxic cost of the regimen was low and patient tolerance remarkably good. This combination chemotherapy can safely be recommended for ambulatory use and does not seem to compromise heavily the patients' quality of life. PMID- 3026239 TI - Purification and properties of DNA gyrase from a fluoroquinolone-resistant strain of Escherichia coli. AB - Subunit A and B proteins of DNA gyrase were separately purified from fluoroquinolone-resistant Escherichia coli GN14181 (MIC of ofloxacin, 100 micrograms/ml) and susceptible strain KL-16. The supercoiling activities of reconstituted Ar+Br (r, resistant) and Ar+Bs (s, susceptible) were 250-fold more resistant to new fluoroquinolones than those of As+Bs and As+Br. PMID- 3026240 TI - In vitro activity of LY146032 against staphylococci, streptococci, and enterococci. AB - The in vitro activities of LY146032 and seven comparative antimicrobial agents against 14 species of staphylococci, streptococci, and enterococci were studied. MICs of LY146032 were less than or equal to 0.5 microgram/ml for all staphylococci, including oxacillin-resistant strains; less than or equal to 0.25 microgram/ml for all streptococci (except viridans group streptococci); and less than or equal to 4 micrograms/ml for all viridans group streptococci and enterococci. MICs were minimally affected by variations in inoculum size, and LY146032 was bactericidal against all species tested. PMID- 3026241 TI - Comparative activity of beta-lactamase inhibitors YTR 830, clavulanate, and sulbactam combined with beta-lactams against beta-lactamase-producing anaerobes. AB - The in vitro activities of the beta-lactamase inhibitors YTR 830, clavulanate, and sulbactam combined with six beta-lactams against 88 beta-lactamase-producing anaerobes were determined. When combined with the beta-lactams, the three beta lactamase inhibitors showed no synergy against the 10 Bacteroides fragilis homology group II strains. When the beta-lactams were combined with the inhibitors, their geometric mean MICs against the remaining 78 strains were reduced from 4.2 to 150.2 micrograms/ml to 0.2 to 12.9 micrograms/ml. The activity of the beta-lactams combined with the beta-lactamase inhibitors was significantly greater than that of the beta-lactams alone against all groups except B. fragilis homology group II, with 76 to 100% of the strains susceptible to ampicillin plus inhibitor and greater than or equal to 90% susceptible to the other combinations. PMID- 3026242 TI - Distribution of porin and lipopolysaccharide antigens on a Pseudomonas aeruginosa permeability mutant. AB - Pseudomonas aeruginosa common surface antigens were compared in a permeability mutant (PCC118) and its parent (PAO503). The distribution of lipopolysaccharide and porin antigens in the mutant supports the conclusion that beta-lactam permeability was affected by lipopolysaccharide-side chain presentation rather than by a change in porin number. PMID- 3026243 TI - Antiviral effect of 9-(1,3-dihydroxy-2-propoxymethyl)guanine against cytomegalovirus infection in a guinea pig model. AB - The antiviral activity of 9-(1,3-dihydroxy-2-propoxymethyl)guanine (DHPG) against guinea pig cytomegalovirus (GPCMV) was evaluated in guinea pig cell cultures and in Hartley guinea pigs. The 50% effective dose of DHPG against GPCMV replication in cell cultures was 71 microM. Ultrastructural studies revealed that DHPG inhibited the formation of viral cores and the production of nucleocapsids, enveloped virions and dense bodies, but the drug did not prevent the formation of virus induced intranuclear tubular structures. In vivo, guinea pigs inoculated intraperitoneally with GPCMV were treated with DHPG, 25 mg/kg subcutaneously, twice daily. Treatment was initiated 24 h after infection and continued for 7 days. During the acute infection, the average body weights of DHPG-treated, virus infected guinea pigs were approximately 14% lower than the sham-treated counterparts on day 10, 11 and 13 post-virus inoculation. Virus infectivity titers were higher in the lungs of DHPG-treated guinea pigs on day 10 than the sham-treated ones. Although there was no significant difference on histopathologic lesions in the spleen, liver and lungs of the drug-treated and the sham-treated guinea pigs, DHPG treated animals appeared to have fewer virus induced lesions or inclusions in the kidneys and salivary glands than the sham treated ones. In addition, virus infectivity titers in the salivary gland of DHPG treated guinea pigs were consistently lower than the sham-treated animals. PMID- 3026244 TI - Comparative efficacy of three 2'-fluoropyrimidine nucleosides and 9-(1,3 dihydroxy-2-propoxymethyl)guanine (BW B759U) against pseudorabies and equine rhinopneumonitis virus infection in vitro and in laboratory animals. AB - The three 2'-fluoropyrimidine nucleosides 1-(2-deoxy-2-fluoro-beta-D arabinofuranosyl)-5-iodocytosine (FIAC), 1-(2-deoxy-2-fluoro-beta-D arabinofuranosyl)-5-iodouracil (FIAU), and 1-(2-deoxy-2-fluoro-beta-D arabinofuranosyl)-5-methyluracil (FMAU), showed high activity in RK13 monolayers against equine rhinopneumonitis virus, (EHV-1, IC50 range 0.02-0.18 microM), Aujeszky's disease virus (SHV-1, pseudorabies, IC50 range 0.25-7 microM) and infectious bovine rhinotracheitis virus (1BR, BHV-1, IC50 range 0.1-3 microM). The activity of these compounds was compared with 9-(1,3-dihydroxy-2 propoxymethyl)guanine (BW B759U, DHPG) in two laboratory animal disease models: EHV-1-induced hepatitis in hamsters and SHV-1-induced encephalitis in mice. All the compounds, provided from 3 to 5 h pre-infection for 5 days, were effective in preventing EHV-1 mortality (at 3-5 mg/kg per day) and in significantly reducing SHV-1 mortality (at 60 mg/kg per day). While FIAU had the greatest activity in vitro, FMAU tended to be more potent in vivo. The reasons for these differences between relative in vitro and in vivo activities are briefly discussed. PMID- 3026245 TI - The poliovirus-induced shut-off of cellular protein synthesis persists in the presence of 3-methylquercetin, a flavonoid which blocks viral protein and RNA synthesis. AB - In poliovirus-infected cells, the viral protein and RNA synthesis were severely reduced, provided 3-methylquercetin was present between 1 and 2 h post-infection. Under these conditions, the virally induced host shut-off remained in effect. On the other hand, in uninfected HeLa cells, protein and RNA synthesis was inhibited only slightly by 3-methylquercetin. The inhibition of poliovirus cytopathogenicity in Vero cells by 3-methylquercetin exhibited a similar time dependence. PMID- 3026246 TI - Photodynamic inactivation of influenza and herpes viruses by hematoporphyrin. AB - Hematoporphyrin (HP), at concentrations as low as 0.5 microgram/ml, was found to inhibit the in vitro replication of influenza A and herpes simplex viruses, but not of several other viruses. The effect required exposure of the viruses or cells to visible light and was demonstrable when HP was administered shortly before virus inoculation or during the infection. In studies on the mechanism of action of HP, we found that in the presence of light, HP caused decomposition of GMP but not of various other nucleosides. It caused breakdown of yeast tRNA and inhibited polymerization of RNA and DNA by influenza virus and HSV-1-specific polymerases as well as some other polymerases isolated from bacterial and mammalian sources. Protective effects of HP and light were demonstrable in embryonated eggs infected with the WSN and PR8 strains of influenza A virus and in mice infected with the WSN strain. HSV-1-induced keratitis in rabbits and HSV 2-induced dermatitis in mice were not responsive to HP treatment. PMID- 3026247 TI - Phosphorylation of nucleoside analogs by equine herpesvirus type 1 pyrimidine deoxyribonucleoside kinase. AB - Replication of equine herpesvirus type 1 (EHV-1) was sensitive to 9-(1,3 dihydroxy-2-propoxymethyl)guanine(DHPG) but relatively resistant to E-5-(2 bromovinyl)-2'-deoxyuridine (BVDU). Likewise, plaque formation by EHV-1 was inhibited by DHPG, but not by BVDU. Plaque formation by a thymidine kinase negative (tk-) mutant of EHV-1 was not inhibited by DHPG. In order to investigate biochemical mechanisms determining the differential sensitivity of EHV-1 to these drugs, the EHV-1-encoded thymidine kinase enzyme activity (TK)1 was partially purified from EHV-1-infected cells and analyzed. The EHV-1-induced enzyme utilized both ATP and CTP as phosphate donors and differed in relative electrophoretic mobility from the TKs of mock-infected and HSV-1-infected cells. Phosphorylation of 3H-dThd by the EHV-1 TK was inhibited by AraT, IdUrd, BVDU, and DHPG. The EHV-1 TK phosphorylated 125I-dCyd and 3H-ACV. The results indicate that EHV-1 encodes a pyrimidine deoxyribonucleoside kinase with broad nucleoside substrate specificity. These observations suggest that the failure of BVDU to inhibit EHV-1 replication is not attributable to an inability of the EHV-1 TK to phosphorylate BVDU, but may result from the incapacity of the viral TK to convert BVDU monophosphate to the triphosphate or from lack of inhibitory effect of BVDU triphosphate on viral DNA polymerase reactions. PMID- 3026249 TI - Comparison of the energetics of lactose active transport: artificial versus enzyme-associated energy source. AB - To further consider the thermochemical method as a useful approach for active transport research and to investigate the characteristic of a proton electrochemical potential (delta mu H+) across the membrane, the energetics of lactose active transport across Escherichia coli membrane vesicles coupled with an artificial electron donor (phenazine methosulfate-ascorbate) has been investigated. The results were compared with those obtained with an enzyme associated electron donor (lactate dehydrogenase-D-lactate). The oxidation of an electron donor provided the energy necessary for the transport process. The observed higher heat of ascorbate oxidation reaction in the presence of a proton ionophore (carbonyl cyanide m-chlorophenylhydrazone) further confirmed the formation of delta mu H+ across the membrane. Part of the oxidation energy was utilized to form delta mu H+. Comparison of the energetics revealed that the magnitudes of delta Hox (the enthalpy of the oxidation reaction) and delta Hm (the enthalpy of the formation of delta mu H+) in the two energy sources were comparable (-46 kcal/mol of ascorbate to -40 kcal/mol of D-lactate for delta Hox and 9.6 kcal/mol of ascorbate to 14 kcal/mol of D-lactate for delta Hm). Comparable and low value (about 1%) was also found in the free energy transfer (defined by delta Gm/delta Gox) from the oxidation reaction to the formation of delta mu H+. These results, in combination with the close values of delta mu H+ observed in the two systems, suggested that the characteristic of the created delta mu H+ was independent of the energy source. Examination of delta Hm might provide the information on the ratio of the number of protons produced, as 1 mol of two different electron donors was oxidized. The oxidation reaction in the presence of membrane vesicles was discussed. PMID- 3026248 TI - Catalysis of superoxide dismutation by manganese aminopolycarboxylate complexes. AB - Complexes of manganese, copper, cobalt, and iron with a variety of aminopolycarboxylates at concentrations from 2 X 10(-7) to 3 X 10(-6) M were tested for superoxide dismutase activity with horse ferricytochrome c as the competing reagent for superoxide. In the presence of excess ligand only manganous nitrilotriacetate and manganous ethylenediaminediacetate showed activity with catalytic rate constants of 2.2 X 10(7) and 1.8 X 10(7) M-1 S-1, respectively, at pH 6, 22 +/- 1 degree C, and 10 mM ionic strength. These rate constants decrease considerably at higher pH. Manganous N-hydroxyethylethylenediaminetriacetate is oxidized by superoxide, but does not appear to have catalytic activity. From the experimental conditions under which the two complexes mentioned above exhibit catalysis, and the inactivity of other metal chelates, it is concluded that an open coordination site is essential but not sufficient to catalyze the dismutation reaction. PMID- 3026250 TI - Purification and characterization of collagenase activator protein synthesized by articular cartilage. AB - We have isolated an activator of collagenase from medium conditioned with articular cartilage. The activity is contained in an acidic protein appearing as a doublet band of Mr 57,000 and 56,000 on sodium dodecyl sulfate polyacrylamide gels. Both components of the doublet have identical isoelectric points as demonstrated by gel electrophoresis. Purified synovial collagenase has a high dependence on the presence of this factor for activity. Other known activators of latent proteolytic enzymes such as trypsin and mercurials will stimulate collagenase but only if activator protein is present. The activator protein is itself a latent metalloprotease because in the presence of p-aminophenylmercuric acetate and calcium it will digest casein. The caseinase activity and collagenase activation activity have identical heat inactivation profiles, both being stable to a temperature of 60 degrees C and partially inactivated at 80 degrees C. The synthesis of the activator is localized in the superficial zone of articular cartilage. PMID- 3026251 TI - Stimulation of the synthesis of collagenase activator protein in cartilage by a factor present in synovial-conditioned medium. AB - We have purified a low molecular weight protein from medium conditioned by calf synovium with physical and biological properties similar to the leukocyte cytokine interleukin 1 (IL-1). The factor is active in stimulating the synthesis (three- to fivefold) of collagenase activator protein (CAP) by the surface (1-2 mm) of articular cartilage while CAP synthesis in the deeper zones of articular cartilage is not affected. Recombinant mouse IL-1 and commercially available purified human IL-1 are also capable of stimulating cartilage to synthesize and secrete CAP. The synthesis of other proteins, including collagenase, appeared to be unaffected by either the synovial factors or the human and mouse IL-1. PMID- 3026253 TI - [Serum neuron-specific enolase (NSE) in patients with small cell lung cancer- comparison with carcinoembryonic antigen (CEA)]. AB - Serum neuron-specific enolase (NSE) was determined by RIA in 102 lung cancer patients. Serum NSE was elevated (greater than 10 ng/ml) in 72% (21 of 29 cases) of small cell lung cancer (SCLC) patients, which was a significantly higher positive rate than those in normal adult controls (0%, 0/48), noncancerous lung disease (17%, 4/24), squamous cell carcinoma (19%, 6/31) and adenocarcinoma (16%, 4/25) (p less than 0.05, respectively). There were no NSE-positive cases in stage I-II lung cancer patients. In SCLC, cases of extensive disease had a significantly higher NSE-positive rate (100%, 8/8) than those of limited disease (62%, 13/21) (p less than 0.05), suggesting that NSE levels were related to the bulk of the tumor. There was an excellent correlation between serum NSE and clinical response. Raised NSE levels were identified significantly more frequently than those of CEA in SCLC before chemotherapy and on relapse (or progression) (p less than 0.025, p less than 0.005, respectively). Thus, serum NSE determinations may be more useful than those of CEA for the staging and monitoring of SCLC. PMID- 3026252 TI - Enzymatic reactions involving orthoarsenate: arsenate is competitive with sulfate in the ATP sulfurylase reaction. AB - In the presence of ATP, Mg2+, and arsenate, ATP sulfurylase from yeast will catalyze the formation of inorganic pyrophosphate. Inorganic pyrophosphate was detected by determination of orthophosphate in the presence of inorganic pyrophosphatase. Two moles of Pi were found for each molecule of ATP in the reaction mixture. The activity of ATP sulfurylase with arsenate as an activating anion was from 1 to 3% of the activity observed with molybdate. PMID- 3026255 TI - [Ifomide]. PMID- 3026254 TI - [A case report of patient with advanced stomach carcinoma of linitis plastica type responding to multi-immunotherapy and chemotherapy]. AB - Multiple-drug (OK-432, PSK and SPG) immunotherapy and chemotherapy provided remission of symptoms for 36 months in a patient aged 21 years suffering scirrhous gastric carcinoma associated with carcinomatous peritonitis in which direct infiltration to the pancreas, retroperitoneum and the left colon was observed. A remarkable improvement with time was observed by endoscopic and roentgenographic observation, and a substantial improvement was also observed in the NK-cell ratios of lymphocyte subsets of the OKT series in relation to immunologic parameters. A tumor necrosis factor [TNF]-like substance was thought to have been induced by multiimmunotherapy in this case. PMID- 3026256 TI - [Role of immune skin reactions in progressive lung cancer during the administration of OK-432--relation to reactions to DNCB, PPD and Su-PS]. AB - In order to study the role of immune skin reactions (DNCB, PPD, Su-PS reaction) in progressive lung cancer, various investigations were conducted, in particular during the administration of OK-432, and the following results were obtained. Before and after chemotherapy, only a slight decrease in the skin reactions was noted. Su-PS reaction was intensified by the administration of OK-432, but other skin reactions were not changed after 3 months, being only slightly intensified by long-term administration. At the time of remission achieved through radiochemotherapy during administration of OK-432, Su-PS reaction was intensified compared to the level before treatment, and a lowering tendency was noted at the time of recurrence. PPD reaction presented a similar tendency, but DNCB reaction did not show this trend. Concerning the relationship between skin reaction and survival period, the positive DNCB reaction group before treatment had a significantly extended survival period compared to that of the negative group. During administration of OK-432, Su-PS reaction was most useful at all timings, while PPD reaction occurred during the intermediate period. Upon observing the Su PS reaction after 3 months of treatment, the prognosis was excellent in cases with erythema of 10 mm or more. However, no such tendency was noted for the PPD reaction. Thus, for understanding the pathologic state and prognosis of patients with progressive lung cancer, the Su-PS reaction was most useful during intra dermal administration of OK-432, the PPD reaction was moderately useful, but the DNCB reaction produced different results. Therefore, for the evaluation of prognosis, it was considered essential to select and combine the skin reactions according to the examination timing and treatment schedule. PMID- 3026257 TI - [Determination of various tumor markers, with special reference to neuron specific enolase in lung cancer]. AB - Serum neuron-specific enolase (NSE) was measured in 23 patients with small cell lung cancer (SCLC) and 184 patients with non-small cell lung cancer (non-SCLC), both of which were untreated. Increased levels of serum NSE were observed in 82.6% (19/23) of SCLC, whereas 9.8% (18/184) of non-SCLC had positive results showing an overall positive rate of 17.9% (37/207) in lung cancer cases. In addition, the elevation of serum NSE levels in non-SCLC patients seemed to suggest poor prognosis. Elevated serum NSE levels returned to normal with either surgical resection of the tumor or response to chemotherapy, after which serum NSE levels were again raised to levels higher than the previous ones in cases of relapse or progression. The evaluation of serum NSE may be a useful marker for both diagnosis and monitoring of responsiveness to therapy as well as for recognition of relapse and progression in SCLC. Identification of NSE as assessed by immunohistochemical procedure employing the ABC method on formalin-fixed paraffin-embedded tissue sections in lung cancer cases of each histological type, showed that some materials from non-SCLC cases were positively stained despite the presence of normal serum NSE levels, and did not always parallel the serum levels. Among other various tumor markers determined, serum CA 19-9 had a relatively high positive rate of 38.2% (42/110) in adenocarcinoma of the lung. PMID- 3026258 TI - Pleural migration of chrysotile fibers after intratracheal injection in rats. AB - After intratracheal injection of 0.5 mg of UICC chrysotile, rats were sacrificed at various time intervals over a 1-month period, and their pleural cavity washed with saline. Samples were examined by means of transmission electron microscopy for identification, quantitation, and sizing of chrysotile fibers. No fibers were found in control rats injected with saline. The size and quantity of fibers that reached the pleural cavity of test rats was time-dependent, with two peaks on day 7 (mean number = 9.6 X 10(4); mean length = 1.3 micron) and on day 21 (mean number = 1.3 X 10(5); mean length = 0.3 micron). Such variations are compatible with a heterogeneous migration of chrysotile fibrils toward the pleura. PMID- 3026259 TI - Hepatic resection for hepatocellular carcinoma. Clinical features and long-term prognosis. AB - Ninety-eight hepatic resections for hepatocellular carcinoma were performed on 94 patients from 1963-1985. HBs antigen was positive in 17% of patients, preoperative serum alpha-fetoprotein was more than 20 ng/mL in 70% of patients, and liver cirrhosis was present in 75% of patients. Hospital mortality rate was 19%, and the volume of operative blood loss was the most decisive factor that affected the short-term prognosis. Excluding the 19 hospital deaths, the long term survival rates of 75 patients were 73%, 42%, and 25% for 1, 3, and 5 years, respectively. Prognostic factors that influenced the long-term prognosis were investigated by comparing the survival curves. Significant differences of survival patterns were noted when analyzed on the basis of preoperative alpha fetoprotein level (less than or equal to 200 vs. greater than 200 ng/mL), tumor size (less than or equal to 5 vs. greater than 5 cm), and tumor capsule. The recurrence of carcinoma was the main cause of death in 56% (42 patients) who died after discharge from the hospital. The development of effective prevention and treatment against recurrent tumors is necessary to improve long-term prognosis. PMID- 3026260 TI - Patterns of failure in patients with resected stage I and II non-small-cell carcinoma of the lung. The Ludwig Lung Cancer Study Group. AB - The pattern of failure was studied in 1012 patients with resected Stage I or II non-small-cell carcinoma of the lung. Initial intrathoracic failure (41%) was more common than initial extrathoracic failure (34%) even though a complete resection was the intent in all patients. The most frequent sites of initial failure were the bronchial resection line (16%) and the central nervous system (CNS) (15%). The site distribution of initial failure does not appear to depend on TNM stage or pattern of nodal involvement. Patients with poorly differentiated disease had a greater rate of initial extrathoracic failure (p less than 0.01), predominantly bone or CNS. Implications for therapy and future research are discussed. PMID- 3026261 TI - Mechanism of anaemia in resistant visceral leishmaniasis. AB - We have studied the mechanism of a transfusion-dependent anaemia found in a two year-old Maltese girl with visceral leishmaniasis that was resistant to multiple courses of antimonial therapy. Major factors contributing to the anaemia were haemolysis occurring in both the massively enlarged spleen and liver and haemodilution resulting from expansion of the plasma volume. There was no evidence of significant ineffective erythropoiesis, but a reduced plasma iron in the presence of greatly increased iron stores suggested that reticuloendothelial hyperplasia was accompanied by abnormal iron retention by macrophages typical of the 'anaemia of chronic disorders'. This may limit the erythropoietic response to anaemia in chronic visceral leishmaniasis. PMID- 3026262 TI - Electrocautery in pulmonary resection. PMID- 3026263 TI - Simultaneous cisplatin fluorouracil infusion and radiation followed by surgical resection in regionally localized stage III, non-small cell lung cancer. AB - Sixty-four patients with stage III (M omicron) non-small cell lung cancer were treated with cisplatin fluorouracil infusion chemotherapy and simultaneous radiation therapy for 5 days every other week. A total of 4 cycles (40 Gy) was followed by attempted surgical resection. Clinical response to the preoperative treatment included 5 (8%) complete and 32 (48%) partial responses. Thirty-nine (61%) underwent the planned operation, and in 9 (23%) of these patients the resected specimens were histologically negative. Clinical assessment failed to predict histological response. With 17 months median follow-up (range, 2.4-29 months), estimated 1-year survival was 61% and median survival was 16 months for all patients. PMID- 3026265 TI - [Viral etiology of diarrheal diseases in Madagascan children]. AB - Our study included a total of 318 diarrheic stools and 52 normal stools collected from out-patients with acute diarrhea at welfare Center and children admitted at Antananarivo City children's Hospital, or control free of infectious disease during 8 months period. Enzyme linked immunosorbant assay and tissue cultures revealed the presence of 152 viral particles (47%) from children with diarrhea and 29 viral particles (55%) from control children. Positive cases were distributed according age, sex, and season factors. The highest infection rate was found in 25-36 months old of the children with diarrhea (72%). The two sexes were equally infected. Enteroviruses were isolated from diarrheic stools with a high frequency (43%) during the rainy and warm season while Rotaviruses were the prevailing agent during the dry and cool season, and Adenoviruses came in second place (19%). In view of our results, the etiological role of these viruses in diarrhea is discussed. PMID- 3026266 TI - [Incidence of rotavirus infections in children with diarrhea in the Majunga region]. AB - Rotavirus were detected by enzyme immuno assay method in 36 p. 100 of faecal specimens from 80 children with acute diarrhoea and in 8 p. 100 of 80 controls, over a period of three months in Majunga area. The incidence of rotavirus was studied according to age group, sex, nutritional status, and clinical aspects. PMID- 3026264 TI - [Enalapril in the treatment of essential arterial hypertension]. AB - This study evaluates the antihypertensive effects of enalapril, a new, potent, long acting angiotensin converting enzyme inhibitor. 69 patients with uncomplicated essential hypertension were included in 5 groups. Group I was used to compare the effects of enalapril and propranolol on blood pressure, renal function, plasma renin activity, aldosterone excretion and plasma lipids in 24 patients after 12 weeks. Group II was used to evaluate long term effects (48 weeks) of these drugs in 13 patients. Group III included 32 patients that received enalapril as monotherapy for 6 to 12 weeks. Group IV was studied to estimate the antihypertensive effect of low doses of hydrochlorothiazide in 18 patients receiving enalapril. Group V was used to compare the antihypertensive effect of hydrochlorothiazide, enalapril or enalapril plus hydrochlorothiazide in 19 patients. The effect on mean blood pressure was similar with enalapril and propranolol (117 versus 103 mmHg and 115 versus 104 mmHg respectively); however, glomerular filtration rate decreased with propranolol (105 versus 87 ml/min; p less than .05) and was unaltered with enalapril (102 versus 98 ml/min). Triglycerides rose with propranolol (179 versus 231 mg/dl; p less than .05) and did not change with enalapril (157 versus 121 mg/dl). In the long term, antihypertensive effects were similar and no significant side effects were observed. In 14/32 patients blood pressure became normal with enalapril alone. Low doses of hydrochlorothiazide (12.5 to 25 mg) decreased mean blood pressure by 10 mmHg when added to enalapril. The antihypertensive effect of enalapril plus hydrochlorothiazide was significant greater than that of enalapril or hydrochlorothiazide alone. Used as monotherapy, enalapril normalised blood pressure in 44% of cases. Addition of low doses of hydrochlorothiazide significantly increased the antihypertensive effect of enalapril. These results show that enalapril is a good antihypertensive agent alone or with low doses of diuretic. PMID- 3026267 TI - [Seroepidemiologic study of cytomegalovirus infections at the maternity department of the Befelatanana General Hospital]. AB - A total of 170 sera from a population of malagasy women aged from 15 to 35 year old were assayed in complement fixing test for CMV-antibodies. 80 p. 100 were found positive. Positive cases have been studied according age groups, matrimonial status, number of gestation. PMID- 3026268 TI - [Viral etiology of acute respiratory infections in Madagascan children]. AB - A total of 80 nasopharyngeal secretions collected from malagasy children (53 boys and 27 girls) with viral acute respiratory infection, aged from 6 days to 10 year old admitted to the Pediatric Department of Antananarivo General Hospital from may to July 1983, were investigated by indirect immunofluorescence method. 54- samples were found positive for respiratory viruses. Distribution according age groups and sex has been studied: children belonging to 25-36 month age group and male sex were more infected. Following viral strains were detected in increasing frequency: Para-influenzae 3: 25 p. 100; RSV: 18 p. 10/; Adenovirus: 18 p. 100; Influenzae A: 13 p. 100; Influenzae B: 9 p. 100; Para-influenzae 1: 5 p. 100. PMID- 3026269 TI - Alterations of myocardial alpha 1-adrenergic receptors in hypertensive cardiac hypertrophy in the rat. AB - The hypertrophied myocardium of 2 kidney-1 clip renal hypertensive rats (2K-1C RHR) displays reduced responsiveness to phenylephrine stimulation. We have studied the effects of hypertrophy on alpha 1-receptor binding properties in three models of hypertension. Specific binding density of an alpha 1-selective antagonist [3H]-prazosin to ventricular membrane of 6 and 10 weeks 2K-1C RHR was significantly decreased (45 +/- 9 fmol/mg protein, n = 14, vs 64 +/- 7.8 fmol/mg protein, n = 20, p less than .001) compared to age matched sham operated control group, but with no change in the total alpha 1-receptor content in the hypertrophied ventricles. The 4 week 1K-1C RHR showed a 29% decrease in alpha 1 adrenergic receptor density with no change in affinity. In contrast, the spontaneously hypertensive rats (SHR) showed an increase in alpha 1-receptors in the prehypertensive stage (4 weeks old) (115.2 +/- 4.9 fmol/mg protein, n = 10, vs 88.7 +/- 12 fmol/mg protein, n = 10, p less than .001) and in the adult stage with established hypertension (15 weeks old) (87.1 +/- 10.5 fmol/mg protein, n = 10, vs 67.3 +/- 9 fmol/mg protein, n = 10, p less than .001) compared to age matched or heart weight matched Wistar Kyoto rats (WKY). Reduced responsiveness to phenylephrine in RHR may be due to reduced density of alpha 1-receptors, but the reason for the increase in alpha 1-receptors in SHR's myocardium remains unclear. PMID- 3026270 TI - Low-dose autologous in vitro opsonized erythrocytes. Radioimmune method and autologous opsonized erythrocytes for refractory autoimmune thrombocytopenic purpura in adults. AB - Adult patients with chronic autoimmune thrombocytopenic purpura (ATP), which proved refractory to various treatments, received a single dose of autologous in vitro opsonized erythrocytes with 100 micrograms of anti-D IgG. In 1983, 30 of these patients were treated with autologous erythrocytes that had been opsonized and labeled with 25 mCi (740 MBq) of technetium Tc 99m; this treatment was designated as the radioimmune method. Favorable responses were noted in 36% of patients so treated. In 1985, another group of 16 patients with refractory ATP received therapy with autologous opsonized erythrocytes (AOPE) and 55% of these patients showed favorable responses. Five (17%) of the patients treated using the radioimmune method attained a complete, long-term (greater than 35 months) remission of their ATP, and five (31%) of the patients treated using AOPE remained in complete remission over 270 days after cessation of therapy. Major complications were not seen. We concluded that the interaction of macrophages with low-dose AOPE is a successful therapeutic approach in ATP refractory to standard treatment. PMID- 3026271 TI - [Comparison of the vasomotor effects of dopamine on the renal and iliac arterial beds of the anesthetized rat]. AB - Dopamine remains the reference in the study of the mechanisms involved in the antihypertensive effects of dopaminomimetics. Its renal vasodilator effects are well characterized but relaxation of other vascular beds is less known. The iliac vascular response to dopamine was studied in the anesthetized rat (pentobarbital) and compared to the renal response. Simultaneous measurements of arterial pressure, iliac and renal blood flows (electromagnetic flowmeter probes, Skalar, Delft) allowed iliac and renal vascular resistance (IVR, RVR) to be calculated. Their variations were studied after intravenous injections of increasing doses of dopamine (1.5 to 200 micrograms/kg) in unpretreated animals and in animals receiving various pretreatments. Without pretreatment, dopamine induced a biphasic renal response, vasodilation partially masked by subsequent vasoconstriction for doses above 12.5 micrograms/kg of dopamine. Simultaneously, IVR was increased. After alpha-adrenolytic pretreatment (prazosin 2.5 mg/kg, i.v.), dopamine decreased the RVR while iliac vasoconstriction persisted. The association of yohimbine (5 mg/kg, i.v.) to prazosin completely abolished the vasoconstrictive effects: dopamine lowered then by about 30% both IVR and RVR. Dopamine-induced iliac and renal decrease in vascular resistance persisted in the presence of a beta-adrenoceptor antagonist [+/-)-sotalol 30 mg/kg, i.v.), after depletion of catecholamines from sympathetic terminals (reserpine 10 mg/kg, i.p. 20 h before the experiment) or inhibition of cyclo-oxygenase (indomethacin 2.5 mg/kg, i.p. 20 h and 1 h before dopamine). On the contrary, a specific antagonist of dopamine receptors, (+)-butaclamol (60 micrograms/kg/min, i.v.) stereoselectivity inhibited the dopamine-induced renal vasodilation but did not modify the iliac response.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3026272 TI - [Regulation of V1-vasopressinergic receptors in the rat]. AB - Using tritiated vasopressin, specific binding sites for vasopressin were identified on splenic membranes of Sprague-Dawley rats. One class of high affinity receptors was characterized with an equilibrium dissociation constant of 1.91 +/- 0.16 nM and a maximal binding capacity of 110 +/- 11 fmol/mg of protein. Several experimental evidences suggest that these receptors belong to the V1 vascular type: The affinity of 8 vasopressin agonists for the receptor is correlated to their vasopressor activity in vivo whereas no such relationship exists when their antidiuretic activity is considered. The affinity of 5 vasopressin antagonists for the receptor is correlated to their antivasopressor activity in vivo whereas no such relationship exists when their antidiuretic activity is considered. Vasopressin does not stimulate cyclic AMP production of splenic membranes. The regulation of these receptors was studied in sodium sensitive (S) and sodium-resistant animals (R) of Dahl receiving a special diet (0.1 or 8% NaCl) for two weeks. In both strains of animals, the number of receptors is smaller in R animals than in S animals, whatever the diet ingested. Splenic membranes of rat bear easily accessible V1-vasopressinergic receptors. These receptors are modulated by sodium intake and the difference in receptor number between S and R rats could explain the increased vascular reactivity of S animals to vasopressin. PMID- 3026274 TI - [Clinical pharmacology of enalapril in hypertension with chronic renal failure]. AB - The effects of a single oral dose of enalapril (20 mg) on blood pressure (BP), heart rate (HR) plasma renin activity (PRA) aldosterone (PA), converting enzyme inhibition (CEI) and enalaprilat (E, active metabolite) were investigated during 96 h in 3 groups of 5 hypertensive patients with (1) normal renal function (creatinine clearance: Clcr greater than 80 ml.min-1); (2) moderate chronic renal failure: 80 greater than or equal to Clcr greater than 30 ml.min-1; (3) severe chronic renal failure: 30 greater than or equal to Clcr greater than 10 ml.min-1. Results are as follows (mean +/- SEM): (Table: see text) CEmax: maximal plasma concentration; TEmax: delay corresponding to CEmax; TE 1/2: plasma elimination half-life; AUCE: area under plasma level versus time curve. a: p less than 0.01; b: p less than 0.001; versus (1). In the 3 groups, CEI reached 87-94% as early as the 3rd h; however, at 96 h, CE1 was higher in (3) than in (1) and (2): 77.6 +/- 3.3% versus 6.0 +/- 1.6 and 17.7 +/- 4.8 (p less than 0.001 respectively). In (3). PRA increased at the 1st h and remained elevated: at 96 h, delta PRA was + 3.0 +/- 2.9 ng.ml-1 -.h-1 in (3) versus + 0.10 +/- 0.06 and + 0.25 +/- 0.17 ng.ml 1.h-1 .n (1) and (2) [(3) versus (1): p less than 0.01]; delta PA was lower in (3): -4.56 +/- 2.01 ng. 100 ml-1 versus -0.54 +/- 0.31 and -2.50 +/- 0.38 ng. 100 ml-1 [(3) versus (1): p less than 0.05].(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3026273 TI - [Peripheral hemodynamic and biological effects of perindopril in the healthy subject. Dose-effect relationship]. AB - The effects of three doses of perindopril (4.8 and 16 mg), a new angiotensin I converting enzyme inhibitor, and of a placebo on systemic blood pressure, heart rate, brachial artery flow and diameter, forearm vascular resistance and the renin angiotensin system biological parameters (plasma converting enzyme activity PCEA, renin activity PRA and aldosterone PA) were compared during a double-blind cross-over study performed in six healthy volunteers. Perindopril dose dependently inhibited PCEA, increased PRA, augmented brachial artery flow and diameter and decreased forearm vascular resistance, whereas it did not affect systemic blood pressure and heart rate. The perindopril-induced increase in brachial artery flow was related (a) at the low dose to the sole arteriolar and (b) at the two highest doses to both arteriolar and large vessels dilatation. Finally, during the ten first hours following drug intake, there was a significant correlation between perindopril-induced PCEA inhibition and brachial artery flow increase. PMID- 3026275 TI - [Antihypertensive action and inhibition of tissue conversion enzyme by ramipril, perindopril and enalapril in the spontaneously hypertensive rat (SHRSP)]. AB - Ramipril and perindopril, the active diacids of two new converting enzyme (CE) inhibitors proved to possess a similar inhibitory potency against rat plasma CE in vitro. These diacid compounds were more active than enalaprilic acid or captopril. In stroke-prone spontaneously hypertensive rats (SHRSP) chronic oral treatment for two weeks with enalapril (30 mg/kg per day), ramipril or perindopril (each 1 mg/kg per day) normalized blood pressure. The CE inhibitor induced changes in parameters of the plasma renin-angiotensin system (angiotensin I, angiotensin II, PRC, CE activity) followed the expected pattern, but were not quantitatively related to the antihypertensive action of the three inhibitors. Four weeks of oral equi-dose treatment with the three CE inhibitors (10 mg/kg per day) inhibited tissue CE activity in various organs including kidney, heart, vascular wall and brain. Ramipril and perindopril lowered blood pressure and tissue CE activity more potently than enalapril. These results confirm the hypothesis that CE inhibition in tissue with subsequent local reduction of ANG II synthesis may contribute to the antihypertensive mechanisms of CE inhibitors. PMID- 3026277 TI - Determination of cytomegalovirus antibody titers in sera of volunteer blood donors by use of complement fixation and two immunoenzymatic tests. AB - A total of 255 serum samples taken from male volunteer blood donors were used for the determination of the level of anti-CMV antibody. Among the serum samples tested by CF, ELISA and EIA procedures, 67,8%, 70% and 68%, respectively, showed the presence of CMV antibodies. PMID- 3026276 TI - [Neonatal myocardial infarct associated with maternal-fetal infection due to Coxsackie B4 virus]. AB - Heart failure with myocardial infarction occurred in the course of a neonatal infection with aseptic meningitis. Coxsackie B4 infection was found in the child and its mother. The unusual myocardial action of the virus, coronary embolus, intermediate role of a pancreatitis. By age 8 months, under digitalis glycosides treatment, growth and psychomotor development were normal. PMID- 3026278 TI - A proline-rich polypeptide (PRP) with immunoregulatory properties isolated from ovine colostrum. Murine thymocytes have on their surface a receptor specific for PRP. AB - A proline-rich polypeptide (PRP) was isolated from ovine colostrum. The polypeptide exhibits immunoregulatory properties. It induces maturation of murine thymocytes into functionally mature helper or suppressor T cells. To elucidate the mechanism of interaction of the polypeptide with thymocytes, effect of PRP immobilized on insoluble carriers on thymocytes was compared with the effect of soluble PRP. It was found that PRP covalently linked to cellulose discs or AffiGel 702 or adsorbed on polystyrene latex beads shows activity similar to activity of PRP in solution. A contact of PRP with the cell surface is satisfactory to induce maturation of thymocytes. PRP adsorbed on polystyrene latex beads forms rosettes with thymocytes. Formation of rosettes is specifically inhibited by soluble PRP. The results obtained suggest that murine thymocytes have on their surface a receptor specific for PRP. PMID- 3026280 TI - Immunohistochemical localization of S100 protein in benign and malignant conditions of the breast. AB - A peroxidase--antiperoxidase technique for S100 protein has been applied to 122 breast lesions from 122 patients. These included 35 cases of fibrocystic disease, 16 cases of sclerosing adenosis, 24 cases of papilloma and papillomatosis, 43 intraduct carcinomas, and four intralobular carcinomas. In fibrocystic disease, S100 protein was demonstrable in large amounts in cells between the duct lining cells and the basement membrane of the ducts, being most pronounced in those exhibiting adenosis. Areas of epitheliosis showed scattered positive cells within the ducts with more strongly positive cells around these ducts. Apocrine metaplasia was moderately positive. No S100 protein was demonstrable in the epithelial lining cells of cysts or within the stroma. In sclerosing adenosis individual cells and groups of cells in the fibrous tissue were strongly positive. In papillomatosis and papilloma, the vascular core and epithelium failed to stain, but a discontinuous layer of cells between the epithelium and basement membrane was positive. In intraduct and intralobular carcinoma the tumor cells were uniformly negative, and wherever fibrocystic disease was also present, S100 protein was variably demonstrable. The study corroborated the view that fibrocystic disease and benign proliferative processes of the breast appear to contain cells that correspond to myoepithelial cells, and suggests that S100 protein may serve as a useful marker in the separation of benign proliferative breast lesions from in situ carcinoma. PMID- 3026279 TI - DNA hybridization for human papillomavirus (HPV) in cervical lesions. Relationship of the presence of various viral subtypes to expression of HPV structural proteins, involucrin, and carcinoembryonic antigen. AB - We studied cervical tissue from 20 patients with a variety of condylomatous, preneoplastic, and neoplastic lesions to detect various subtypes of human papillomavirus (HPV) at the molecular level by DNA hybridization. In addition, mirror image biopsy specimens were studied by an immunoperoxidase technique for the presence of HPV structural antigens, carcinoembryonic antigen (CEA), and involucrin, as markers of disturbed maturation and/or neoplasia. Of the 20 patients studied, we were able to demonstrate the presence of either HPV 16 or 18, or both, in three of seven squamous carcinomas tested, and three of five dysplasias. Interestingly, two cases of squamous carcinoma and one moderate dysplasia demonstrated HPV types 6 and 11 concurrently with HPV 16 and/or 18. Four of six condylomas showed HPV subtypes 6/11, and only one condyloma showed weak hybridization with HPV 18. Human papillomavirus structural antigen was seen in three of eight condylomas, and three of five dysplasias, but not in any carcinoma. All condylomas showed intense staining for involucrin in full thickness of the epithelium, but the high-grade dysplasias and carcinomas showed only focal or absence of staining for involucrin. Carcinoembryonic antigen was expressed in 50% of the carcinomas with a pattern similar to that seen with involucrin, but did not correlate with any particular subtype of HPV. The molecular hybridization method seems to be superior for the detection of HPV lesions, and possibly for the prediction of their biologic behavior. PMID- 3026281 TI - Hepar lobatum associated with chemotherapy and metastatic breast cancer. AB - Hepar lobatum is now a rarity in developed countries. In this article, we describe three cases with typical liver morphology associated with carcinoma of the breast metastatic to the liver, treated with combination chemotherapy. The pathogenesis of hepar lobatum in our cases is multifactorial with tumor-related desmoplasia and effects of chemotherapy, with resultant tumor necrosis and fibrosis playing a major role. PMID- 3026283 TI - [Methodologic studies of ileal flow measurement using large lumen intestinal cannulae in swine]. AB - A special operation method and intestinal cannulae were developed for the ascertainment of the ileal nutrient digestibility of roughage and root crops. The presented method has proved reliable over a period of 2 years. The possibility of ascertaining the chyme flow directly (by total collection) or indirectly (with an inert marker) is described. The reduction of the expenditure of labour by cutting down the collecting period is suggested for routine studies. PMID- 3026282 TI - Physiology and metabolism in isolated viral septicemia. Further evidence of an organism-independent, host-dependent response. AB - The hypothesis has been advanced that the human systemic septic response is a function of the host and not of the type of infecting organism. Metabolic and physiologic data from five immunosuppressed transplant recipients with isolated cytomegaloviral sepsis and viremia were prospectively evaluated. Serial cultures obtained from lung, sputum, urine, wound, blood, and invasive lines were positive for virus and negative for bacterial or fungal pathogens. The results were compared with two data banks derived from either victims of multiple trauma without sepsis or surgical patients with early bacterial or fungal sepsis. Statistically significant differences between the patients and the nonseptic reference group were noted for cardiac index, total peripheral resistance, arteriovenous oxygen content difference, oxygen consumption, and levels of triglycerides, proline, phenylalanine, tyrosine, alpha-aminobutyrate, and alanine. No such differences were present for these data compared with the septic reference group. Physiologic data obtained just before death in three patients indicated a failure of oxygen transport. It appears that the systemic septic response to viral agents is indistinguishable by physiologic and metabolic criteria from that resulting from bacterial or fungal agents. PMID- 3026284 TI - Effect of defaunation and refaunation of the rumen on rumen fermentation and N flow in the duodenum of sheep. AB - In order to confirm earlier fragmentary results, the effect of defaunation and refaunation of the rumen on the fermentation pattern and flow of N-components in the proximal duodenum of two sheep was investigated. Defaunation had no effect on acetic acid as a proportion of the total volatile fatty acids in the rumen, while the proportions of propionic acid increased with a concomitant decrease in butyrate. Refaunation resulted in lower acetic acid and higher butyric acid proportions. The concentration of ammonia N in the rumen was clearly decreased after defaunation, already indicating an effect of the elimination of protozoa on nitrogen metabolism in the rumen. Defaunation also increased significantly the flow of total N, non ammonia N and individual and total amino acids in the proximal duodenum. Defaunation resulted in higher bacterial growth efficiency, significantly in one sheep, but the decrease after refaunation was statistically significant for both sheep. Determination of rumen digestibility of organic matter and acid detergent fibre (ADF) revealed lower values in the absence of the protozoa, while total digestibility was only influenced to a much lower extent. This indicated a shift of digestion from rumen to the lower digestive tract. Finally, earlier work is discussed in the light of the present findings. PMID- 3026285 TI - [Nitrogen metabolism in the large intestine of ruminants. 3. Microbial utilization of intracecally administered 14C- and 15N-marked urea in the large intestine of sheep in simultaneous intracecal administration of partially hydrolyzed straw meal]. AB - Two experiments were performed on sheep, receiving on maintenance level a pelleted straw ration high in crude fibre (straw, 70.5%; dried sugar beet pulp, 12%; cereals, 10%; urea, 2%; ammonium hydrogen carbonate, 3%; minerals 2,5%). The animals were fitted with ileo-caecal re-entrant cannulas. The effects of the introduction of HC1-partly hydrolysed straw meal into the digesta of the large intestine on the digestion processes in that segment were studied. Under these conditions the metabolism of 14C and 15N labelled urea, which was given into the caecum, was estimated. In experiment 1 (E 1; 2 animals) unlabelled, precollected digesta were hourly reintroduced together with 14C and 15N labelled urea via the caecal cannula. In experiment 2 (E 2; 3 animals) the digesta were supplemented with partly hydrolysed straw meal (10% of the mean daily DM-intake with the ration). The supplement of partly hydrolysed straw meal caused an increase of the 15N excretion with faeces from 13.4% (E 1) to 19.8% (E 2) of the dose. The 15N was mainly incorporated in the bacterial fraction (98% E 1; 96% E 2). As a reason for the increased 15N incorporation into the bacterial fraction of 106.4 mg15N' in E 2 vs. 67.3 mg15N' in the experiment without straw meal supplement the higher supply of energy as fermentable carbohydrates was assumed. PMID- 3026287 TI - Three different forms of tubular structures associated with the replication of bovine rotavirus in a tissue culture system. Brief report. AB - We demonstrated by electron microscopy that three different forms of tubular structures are present both within infected cells and in cultured fluid of bovine rotavirus in a tissue culture system. PMID- 3026286 TI - Analysis of immediate-early and early proteins of murine cytomegalovirus in permissive and nonpermissive cells. AB - The immediate-early (IE) and early proteins induced by murine cytomegalovirus (Smith strain) in permissively infected 3T3-L1 murine fibroblasts were identified by SDS polyacrylamide gel electrophoresis. Ten proteins were classified as IE by their time of synthesis and by their synthesis in the presence of actinomycin D following reversal of a cycloheximide mediated protein synthesis block. By exclusion, seven proteins were classified as early class. Eleven of these proteins were precipitated by MCMV antiserum. The role of IE and early proteins in the replication of the virus was studied by infection of a murine macrophage cell line (J 774A.1) and human foreskin fibroblast (HFF) cells. The infections were characterized as nonpermissive by several criteria, including lack of production of infectious virus or viral DNA. However, the major IE and early MCMV proteins were detected in the nonpermissively infected cells. The block to virus replication in the macrophage and human fibroblast cells appeared to occur after the switch from IE to early protein synthesis, but before viral DNA replication. PMID- 3026288 TI - Intracellular localization and transport of three different bovine herpesvirus type 1 glycoproteins involved in neutralization. AB - Monoclonal antibodies against 3 different glycoproteins of bovine herpesvirus type 1 (BHV-1) involved in virus neutralization were used in indirect immunofluorescence (IIF) tests to characterize the appearance and transport to the plasma membrane of virus antigens in the infected cells. Antibodies against gp 117 and gp 71 glycoproteins first showed pronounced ring-like nuclear fluorescence at 4 hours post-infection (PI), followed by staining of the perinuclear region, presumably the Golgi apparatus. In contrast, antibody against gp 87 produced staining in cell-to-cell junctional areas at 3 hours PI before any staining close to the nucleus. The expression of the 3 glycoproteins at the surface of the infected cells was confirmed by the use of monoclonal antibodies having neutralizing activity, but not by non-neutralizing antibodies against gp 117 and gp 71. Non-neutralizing antibody against gp 87 detected the surface fluorescence only in those cells showing marked degeneration. Inhibition of glycosylation of the viral glycoproteins with tunicamycin (TM) was followed by interference with transport of gp 117 and gp 87 to the plasma membrane. On the other hand, gp 71 was incorporated into the plasma membrane despite the lack of N linked glycosylation. PMID- 3026290 TI - The role of epithelial cell differentiation in the expression of herpes simplex virus type 1 in normal human oral mucosa in culture. AB - We have examined by immunofluorescent antibody staining technique the expression of herpes simplex virus type 1 (HSV-1) in organ cultures of the normal human oral mucosa. The expression of HSV-1 antigen was found selectively in the epithelial cell layers in relatively undifferentiated states such as basal layer and lower prickle cell layer in addition to the basement membrane. When the epithelial cells dissociated from the oral mucosa were infected with HSV-1 and association of the HSV-1 expression with the cellular differentiation was examined, the epithelial cells containing laminin in an undifferentiated state were permissive for the expression of HSV-1 antigen whereas terminally differentiated epithelial cells with the cornified envelope did not express HSV-1 antigen. These findings indicate that the expression of HSV-1 antigen is restricted in the mucosal epithelial cells in a differentiated state, although the possibility that the cornified envelope might protect the cells from infection is not excluded. PMID- 3026289 TI - Analysis of molecularly cloned DNA reveals minor differences among three virus strains of Aleutian disease of mink parvovirus. Brief report. AB - Molecular clones representing a 1.55 kbp genomic segment from three strains of Aleutian disease parvovirus (ADV) were studied. All three clones directed synthesis of viral structural antigens. In addition, 19 of 23 restriction sites were shared among viruses. PMID- 3026291 TI - Aujeszky's disease vaccination and infection of pigs with maternal immunity: effects on cell- and antibody-mediated immunity. AB - SCC, ADV-SCC, ADV-ADCC and ADV-LYST as well as ND50-titres of neutralizing serum antibodies were examined in 36 passively immune pigs, 25 of which were vaccinated at 3 weeks of age and partly revaccinated 3 weeks later. Twenty-five vaccinated animals and 8 non-immune control pigs were challenged with infectious ADV. Independent of the state of maternal immunity the cytotoxic response of the white blood cells from all the animals was low at WPP 3 but rose with increasing age. ADV-LYST occurred only in some of the animals. A single vaccination evoked no significant effect on our immune parameters, but revaccination led to higher ADV LYST and ADV-ADCC. In pigs vaccinated at WPP 3 the neutralizing serum titres decreased gradually, similar to unvaccinated animals, indicating that the antibodies were of maternal origin. However, after vaccination at WPP 6, no further decline of ND50-titres could be detected, pointing to a limited antibody production. Animals vaccinated at WPP 3 and revaccinated 3 weeks later showed a significant increase of serum neutralizing titres. Whereas the controls showed typical symptoms of Aujeszky's disease, the immune animals, especially the unvaccinated passively immune pigs, showed only elevated temperatures and most of them excreted small amounts of ADV. The development of cellular immunity after infection was rather similar within the maternally immune group independent whether the animals had been vaccinated or not, but ADV-ADCC and ADV-LYST showed a more rapid progress within the vaccinated group than in the non-vaccinated group and the non-immune control group. Infection resulted in significantly higher ND50 titres in vaccinated and revaccinated animals than in unvaccinated animals, indicating a secondary response in those pigs. Thus, ADV sensitization of lymphocytes had been evoked by vaccination despite the presence of maternal antibody. The interpretation of the results was complicated by great individual and litter-dependent variations of the immune parameters. PMID- 3026293 TI - Beta-adrenergic receptors in human trabecular meshwork. PMID- 3026292 TI - Peroxidase release from rat submandibular salivary acinar cells in vitro. AB - Acinar cell aggregates were isolated and then incubated for 60 min with or without isoproterenol, phenylephrine, carbamylcholine, and insulin, and peroxidase activity in the cells and medium assayed. Isoproterenol stimulated the release of 70-80 per cent of total peroxidase activity. Carbamylcholine stimulation resulted in the release of only 15 per cent of this activity; phenylephrine had no effect. Insulin alone failed to stimulate peroxidase secretion, but it potentiated the response of the cells to carbamylcholine. PMID- 3026294 TI - Effects of enalapril and hydrochlorothiazide on blood pressure, renin-angiotensin system, and atrial natriuretic factor in essential hypertension: a double blind factorial cross-over study. AB - The hypotensive and hormonal effects of the angiotensin converting enzyme (ACE) inhibitor enalapril (10 mg twice daily) were compared with those of hydrochlorothiazide (25 mg twice daily), with the two drugs in combination and with placebo in 21 patients with essential hypertension. For each patient there were four randomised double-blind treatment phases, each of four weeks' duration, which comprised a 2 X 2 factorial experiment. All blood pressure parameters were reduced in the three active treatment phases compared to placebo (p less than 0.001). Supine mean blood pressures were 119 mmHg (placebo), 113 mmHg (hydrochlorothiazide), 108 mmHg (enalapril), and 98 mmHg (hydrochlorothiazide plus enalapril) (SEM 3 mmHg, ANOVA). Enalapril and hydrochlorothiazide were equally effective and well tolerated and their hypotensive effects were additive. Enalapril increased plasma renin activity (PRA), reduced plasma angiotensin II (AII) and aldosterone concentrations, and reduced ACE activity, whereas hydrochlorothiazide increased PRA, plasma AII, and aldosterone concentrations without altering ACE activity. With combination treatment the effects of enalapril on PRA and plasma AII concentrations were potentiated whereas those on plasma aldosterone concentration and ACE activity were additive. Atrial natriuretic factor plasma concentration in the placebo phase was 92 pg/ml and increased to 145 pg/ml in the hydrochlorothiazide phase (p less than 0.001, SEM 13 pg/ml), but there was no significant change in either the enalapril or combination phases. PMID- 3026295 TI - Adult onset Still's disease or coxsackie polyarthritis? AB - The clinical manifestations and laboratory findings of two patients with a presumptive diagnosis of coxsackie B4 virus infection are described. A striking feature was the similarity with adult onset Still's disease, with spiking fever, evanescent macular rash, and severe polyarthritis. This latter feature persisted for many weeks and required steroids to control the symptoms. Review of the literature has supported the proposition that many cases of adult onset Still's disease may be due to coxsackie B4 or other viral infection and it is suggested that these agents should be actively sought in future cases. PMID- 3026296 TI - Inclusion body hepatitis associated with adenovirus-like particles in a cockatiel (Psittaciformes; Nymphicus hollandicus). PMID- 3026297 TI - Comparison of two gel diffusion precipitin tests in the serodiagnosis of caprine arthritis encephalitis virus infection in goats. PMID- 3026298 TI - Characterisation of Histophilus ovis and related organisms by restriction endonuclease analysis. AB - The banding profiles generated by Bam H1 restriction endonuclease cleavage of bacterial DNA from clinical and reference isolates of Histophilus ovis, Haemophilus somnus and related bacteria were compared. H. ovis, H. somnus and Haemophilus agni isolates were found to have distinct similarities in banding profiles characterised by 10 common bands between 2.0 and 9.6 kilobases (kb). The close taxonomic relationship of these isolates was reinforced by these findings. The reference isolates examined in this study--Actinobacillus lignieresii, Actinobacillus seminis, H. agni, H. somnus, H. ovis, Haemophilus influenzae, Haemophilus parainfluenzae, Haemophilus parahaemolyticus--could be distinguished from each other on the basis of their characteristic banding profiles. Actinobacillus sp were observed to have more bands between 2 and 23 kb compared with the H. ovis and Haemophilus sp isolates studied. Analysis of isolates from an experimental infection trial illustrated the potential of restriction endonuclease analysis in molecular epidemiological applications. It was possible to demonstrate by this means that the post-challenge isolates had identical banding profiles to the challenge (or infecting) isolate which had a distinctly different banding profile from that of pre-challenge H. ovis isolates. Furthermore, restriction endonuclease analysis of H. ovis isolates obtained from follow-up investigations of a recurrent problem of epididymitis in unmated rams, indicated that the H. ovis isolates implicated in epididymitis, were present as a single strain in a number of sheep over a period of time. This suggested that the mechanism of transmission was by perinatal perputial contamination. PMID- 3026299 TI - Identification of uroliths by infrared spectroscopy. AB - Wet chemical tests have deficiencies when applied to mixtures containing silica, which are common in the uroliths of some domestic animals. Consequently, the applicability of an infrared spectroscopic method was tested on 104 uroliths obtained from cattle, sheep, goats, horses, pigs, dogs, a chicken and a rabbit during diagnostic investigations. The following components were satisfactorily identified: silica, calcium oxalate, calcium carbonate, calcium phosphate, magnesium ammonium phosphate, magnesium phosphate and urates. The infrared characteristics of these compounds and their mixtures are described. PMID- 3026300 TI - Development of an enzyme linked immunosorbent assay (ELISA) for the detection of bovine serum antibody to bovine viral diarrhoea virus. AB - An enzyme linked immunosorbent assay (ELISA) was developed to detect antibody to bovine viral diarrhoea virus (BVDV) in bovine serum. The ELISA results were compared with those of the serum neutralisation test (SNT) using serums from 6 experimentally infected calves bled at intervals from 0 to 154 days postinfection and 886 field samples. The optical density (OD) produced by a single dilution of test serum was compared with a standard curve and the result expressed in ELISA units. Despite wide variation between absolute ELISA and SNT results, an agreement of 97% was obtained when reciprocal SNT titres greater than or equal to 8 and ELISA units greater than or equal to 10 were taken as indicative of a specific reaction. The ELISA was shown to be an efficient method of measuring antibody in bovine serum samples and would assist in any large scale screening of cattle herds for BVDV antibody. PMID- 3026301 TI - Encephalomyocarditis virus disease of pigs associated with a plague of rodents. AB - An epizootic of encephalomyocarditis virus (EMCV) disease in pigs in the central west of New South Wales in association with a plague of mice (Mus musculus) in 1984 is described. The disease was confirmed in 47 outbreaks in 37 piggeries and 1152 pigs died, representing an overall death rate of 17.4% in pigs considered at risk. The disease was diagnosed in both intensively housed pigs and pigs farmed outdoors, with mortality rates higher in piggeries with less than 50 sows. The age at which pigs died ranged from 4 days to 24 weeks with higher death rates in younger pigs. Serological testing of pigs slaughtered at Blayney abattoir indicated EMCV infection to be more widespread than the disease reported. Mice were present in all piggeries reporting the disease while rats were present in 66% of the outbreaks. The role of rodents as natural reservoirs of EMCV and the possibility of variations in pathogenicity amongst strains of the virus are discussed. PMID- 3026302 TI - The application of the ELISA to the diagnosis and control of avian encephalomyelitis. AB - An ELISA for measuring serum antibody against avian encephalomyelitis virus (AEV) was evaluated for its application to the diagnosis and control of avian encephalomyelitis (AE). A scoring system was developed for this ELISA (AE ELISA Index) so that the overall level of antibody in the flock could be presented in a single, convenient number. During suspected outbreaks of disease thought to have been caused by AEV infection, the AE ELISA-Index increased in sequential serum samples. High levels of antibody against AEV were measured in 13 flocks experiencing egg productivity problems. Variable levels of antibody activity against infectious bronchitis virus (IBV) were also observed in 11 of these flocks. The AE ELISA-Index was correlated with the embryo susceptibility test. Application of the AE ELISA has indicated that natural exposure to the virus does not occur in all flocks, and vaccination failures were detected sufficiently early for revaccination to be administered before the onset of lay. PMID- 3026303 TI - Plasma melatonin concentrations in depression. AB - The pineal hormone melatonin was measured in midnight plasma samples from 11 depressed patients and 18 control subjects. Plasma melatonin concentrations were significantly lower in the depressed patients. The implication of these findings for beta-adrenoceptor subsensitivity in depression is discussed. PMID- 3026306 TI - Analysis of a large nontranscribed spacer in the ribosomal DNA of the house cricket, Acheta domesticus (Orthoptera:Gryllidae). AB - An analysis of a 29-kilobase nontranscribed spacer fragment in the ribosomal DNA (rDNA) of the house cricket, Acheta domesticus, revealed a highly repetitious structure. A total of eight EcoRI repeats of three different size classes measuring 259, 420, and 508 base pairs (bp) was mapped to a region 2 kilobases (kb) from the 18 S coding region. The repeats were oriented in a nonrandom manner and had sequences homologous to DNA located immediately adjacent to the repetitive array. DNA sequence analysis showed that the repetitive region was composed of smaller direct repeats 66, 67, and 383 bp in length. There was minor length heterogeneity of the chromosomal restriction fragments containing the entire array, indicating that a variable number of EcoRI repeats is a minor contributor to the total repeat-unit length heterogeneity. Immediately upstream from the EcoRI array there is a 17-kb region composed of 50 to 60 subrepeat elements recognized by a variety of restriction endonucleases. A subcloned SmaI repeat from the array was not homologous to any other part of the rDNA repeat unit or other chromosomal DNA. There was little length heterogeneity in restriction fragments containing the chromosomal 17-kb repetitions region. Immediately upstream from the 17-Kb region there is a 4.1-kb segment with sequences homologous to the EcoRI repeats. PMID- 3026305 TI - Variation in ten lysosomal hydrolase enzyme activities in inbred mouse strains. AB - Activities of 10 lysosomal hydrolase enzymes (beta-hexosaminidase, beta galactosidase, alpha-galactosidase, alpha-mannosidase, beta-mannosidase, alpha-L fucosidase, beta-glucuronidase, alpha-glucosidase, alpha-N acetylgalactosaminidase, and acid phosphatase) were determined in eight organs (brain, liver, kidney, spleen, heart, skeletal muscle, lung, and testis) in males and females of six inbred mouse strains (C57BL/6J, C3H/HeJ, DBA/2J, BALB/cJ, P/J, and 129/J). Examples of enzyme-specific variation, organ-specific variation, and enzyme- and organ-specific variation were found. New enzyme-specific variants with the features of systemic regulators for alpha-L-fucosidase and beta mannosidase were found. Known variants were detected. Organ-specific variants had some of the properties expected for a new class of genes affecting multiple enzymes: organ-specific regulators. PMID- 3026307 TI - Characterization of multiple forms of phosphoinositide-specific phospholipase C purified from human platelets. AB - The origin and physiological significance of the multiple Mr forms of phosphoinositide-specific phospholipase C in human platelets were investigated. The higher-Mr (400,000 and 270,000) forms of the phospholipase C were converted into the 100,000-Mr form without substantial loss of activity by incubation with a Ca2+-dependent proteinase partially purified from human platelets. These three forms of the phospholipase C were purified approx. 200-500-fold from outdated human platelet supernatants. SDS/polyacrylamide-gel electrophoresis and gel filtration analysis suggested that the higher-Mr forms of phospholipase C were complexes of 140,000-Mr subunits, whereas the lower-Mr form consisted of a single 95,000-Mr subunit. The substrate specificity of the purified phospholipase C was investigated by using 32P-labelled polyphosphoinositide substrates purified from human platelets by a new method utilizing h.p.l.c. on an amino column. Activity against all three phosphoinositides was detected at micromolar concentrations of Ca2+; this hydrolysis was markedly stimulated by phosphatidylethanolamine and inhibited by phosphatidylcholine. Comparison of the different forms of purified phospholipase C revealed no major differences in Ca2+-sensitivity or substrate specificity. Thus, although the suggestion that the high-Mr forms of human platelet phosphoinositide-specific phospholipase C were converted into a lower-Mr form by a Ca2+-dependent proteinase has been substantiated, the physiological significance of this process remains to be determined. PMID- 3026308 TI - A spectrophotometric assay for superoxide dismutase activities in crude tissue fractions. AB - A sensitive and reliable assay method was developed to characterize crude cell homogenates and subcellular fractions with regard to their superoxide dismutase (SOD) activities. The determination of SOD activities was based on the well-known spectrophotometric assay introduced by McCord & Fridovich [(1969) J. Biol. Chem. 244, 6049-6055], with partially succinylated (3-carboxypropionylated) rather than native ferricytochrome c as indicating scavenger. Partial succinylation of cytochrome c resulted in minimization of interference associated with the interaction of cytochrome c with mitochondrial cytochrome c oxidase or cytochrome c reductases. The further increase in specificity, with regard to exclusion of cytochrome c oxidase interference, gained as a consequence of the high pH of 10 enabled the analysis of samples as rich in cytochrome c oxidase activity as the mitochondrial fraction in the presence or absence of membrane-disrupting detergents. Linear relationships for the dependence of the SOD activities with protein concentration were obtained with rat liver homogenate, mitochondrial and microsomal fractions, indicating negligible interference. Furthermore, by choosing a high pH for the assay medium, a 4-fold increase in sensitivity compared with the classical SOD assay, carried out at pH 7.8, was gained as well as a more precise resolution of Cu/Zn-SOD and Mn-SOD by 2 mM-KCN in samples with a high ratio of Mn-SOD to Cu/Zn-SOD, such as mitochondria. The complete trapping of the O2.- radicals, which was more feasible at pH 10 than at pH 7.8, enabled the application of a simple equation derived for the calculation of appropriately defined units of SOD activity from a single experiment. PMID- 3026304 TI - Platelet activation in normo- and hyperlipoproteinemias. AB - In the last few years it became obvious that platelets are involved in the development of atherosclerotic diseases. This involvement of platelets has been taken into account in the "response to injury" hypothesis of atherosclerosis. The hypothesis is based on the assumption that atherosclerotic lesions result from endothelial injury, followed by the interaction of vessel wall constituents with lipoproteins, macrophages, and platelets. In the first part of this review, general aspects of platelet activation are summarized and the pathways of platelet aggregation as well as their involvement in blood coagulation are discussed. The second part of this paper describes the influence of cholesterol, lipoproteins, and apolipoproteins upon the activation and metabolic behavior of platelets. Physiological and pathophysiological processes particularly occurring in different types of hyperlipoproteinemias and atherosclerotic disorders are discussed in this context. PMID- 3026309 TI - Thyroliberin action in pituitary cells is not inhibited by pertussis toxin. AB - The effects of pertussis toxin on the responses of rat pituitary-tumour (GH) cells to thyrotropin-releasing hormone (thyroliberin, TRH) were examined. Treatment of cells with pertussis toxin did not alter the affinity or concentration of TRH receptors, or the sensitivity of the TRH receptor to inhibition by guanine nucleotides. TRH caused an increase in low-Km GTPase activity in membrane-containing fractions from both control and pertussis-toxin treated cells. TRH stimulation of inositol phosphate formation was insensitive to pertussis toxin. TRH caused a biphasic increase in the concentrations of cytosolic free Ca2+ as monitored by intracellularly trapped Quin 2, and this increase was the same in control and toxin-treated cultures. The toxin did not alter the increase in prolactin and growth-hormone (somatotropin) release stimulated by TRH or shift the TRH dose-response curve, and it did not affect the TRH-induced rise in prolactin synthesis measured over 24 h. However, pertussis toxin did block the ability of somatostatin and muscarinic agonists to inhibit prolactin and growth-hormone secretion stimulated by vasoactive intestinal peptide when analysed under the same conditions as those in which the TRH system was unaffected. These data indicate that the guanine nucleotide effects on TRH binding and activity are not mediated by Ni, but possibly by another member of the family of guanine-nucleotide-dependent regulatory proteins. PMID- 3026310 TI - Effects of dehydrouramil on protein phosphorylation and insulin secretion in rat islets of Langerhans. AB - Dehydrouramil hydrate hydrochloride (DHU), a stable analogue of alloxan, inhibited the phosphorylation of an endogenous protein of Mr 53,000 catalysed by a Ca2+-calmodulin-dependent protein kinase in extracts of islets of Langerhans. The concentration of DHU required for 50% inhibition was 0.09 mM. DHU did not inhibit islet cyclic AMP-dependent protein kinase and caused only slight inhibition of Ca2+-phospholipid-dependent protein kinase. Inhibition of Ca2+ calmodulin-dependent protein kinase was neither prevented nor reversed by dithiothreitol. DHU did not affect the ability of calmodulin to activate cyclic AMP phosphodiesterase. In intact islets, pre-exposure to DHU impaired the insulin secretory response to glucose and blocked the potentiatory effect on insulin secretion of forskolin, an activator of adenylate cyclase, and of tetradecanoylphorbol acetate (TPA), an activator of Ca2+-phospholipid-dependent protein kinase. The increase in islet cyclic AMP elicited by forskolin was not affected by DHU. The data are consistent with the hypothesis that protein phosphorylation catalysed by a Ca2+-calmodulin-dependent protein kinase may play a central role in the regulation of insulin secretion. PMID- 3026311 TI - Conformational transitions in the Ca2+ + Mg2+-activated ATPase and the binding of Ca2+ ions. AB - We have studied the fluorescence of the Ca2+ + Mg2+-activated ATPase of sarcoplasmic reticulum labelled with fluorescein isothiocyanate. The change in intensity of fluorescein fluorescence caused by addition of Ca2+ to the labelled ATPase can be interpreted in terms of a two-conformation model for the ATPase, one conformation (E1) having a high affinity for Ca2+, the other (E2) a low affinity. Effects of Ca2+ as a function of pH allow an estimate of the effect of pH on the E1/E2 ratio, consistent with kinetic studies. A model is presented for binding of Ca2+ to the ATPase as a function of pH that is consistent both with the data on the E1/E2 equilibrium and with literature data on Ca2+ binding. PMID- 3026313 TI - Catalytic properties of a purified phosphatidylinositol-4-phosphate kinase from rat brain. AB - A phosphatidylinositol-4-phosphate (PIP) kinase activity was purified from rat brain extract through several chromatographic steps to yield an active preparation (specific activity 1 mumol of 32P incorporated into phosphatidylinositol 4,5-bisphosphate/min per mg of protein) with an apparent molecular size of 100-110 kDa in the native form. The isolated PIP kinase required Mg2+ (optimally 20-30 mM) for its activity and was not influenced by Ca2+. The enzyme used ATP (Km 25 microM) and GTP (Km 133 microM) as phosphate sources and appeared specific for PIP (Km 3.3 micrograms/ml) as the lipid substrate. The PIP-phosphorylation reaction was inhibited by micromolar concentrations of heparin [ID50 (concn. giving 50% inhibition) 2 micrograms/ml] and the flavonoid quercetin (ID50 0.2 microM). Whereas heparin behaves as a competitive inhibitor to PIP, quercetin was competitive towards ATP (or GTP). Phosphorylation of the preparation by a highly active purified protein kinase C did not detectably alter PIP kinase activity. Whereas 12-O-tetradecanoylphorbol acetate and various phospholipids had no effect, phosphatidylserine elicited a dose-dependent activation of PIP activity. This suggests that a phosphatidylserine-PIP kinase interaction may be considered as a possible regulatory process at the cell-membrane level. PMID- 3026312 TI - Effect of thrombomodulin on the kinetics of the interaction of thrombin with substrates and inhibitors. AB - Thrombomodulin decreased by 20-30% the Michaelis constant of two tripeptidyl p nitroanilide substrates of thrombin. Thrombomodulin increased the rate of inactivation of thrombin by two peptidyl chloromethane inhibitors by a similar amount. This effect appeared to be due to a decrease in the dissociation constants of the inhibitors. An improved method for the separation of fibrinopeptides A and B by h.p.l.c. was developed, and this method was used to study the effect of thrombomodulin on the thrombin-catalysed cleavage of fibrinogen. In this reaction, thrombomodulin was a competitive inhibitor with respect to the A alpha-chain of fibrinogen. The release of fibrinopeptide B was also inhibited by thrombomodulin. Analysis of the inhibition caused by thrombomodulin with respect to fibrinopeptides A and B yielded the same dissociation constant for the thrombin-thrombomodulin complex. In the presence of thrombomodulin, the rate of inactivation of thrombin by antithrombin III was stimulated 4-fold. This stimulation showed saturation kinetics with respect to thrombomodulin. Thrombomodulin was found to compete with hirudin for a binding site on thrombin. As a result of this competition, hirudin became a slow-binding inhibitor of thrombin at high thrombomodulin concentrations. Estimates of the dissociation constant for thrombomodulin were obtained in several of the above experiments, and the weighted mean value was 0.7 nM. PMID- 3026314 TI - Protein kinase C and an endogenous substrate associated with adenohypophyseal secretory granules. AB - Secretory granules isolated from anterior pituitary glands were examined for Ca2+/phospholipid-dependent protein kinase (protein kinase C) activity as well as the occurrence of granule-associated substrate proteins. Sheep adenohypophyses were fractionated by differential and sucrose-density-gradient centrifugation to yield a granule fraction enriched for luteinizing-hormone (lutropin)-containing secretory granules. Marker-enzyme analysis showed no detectable cytosolic contamination, although there were small amounts of plasma membranes (2-4%) and lysosomes (4-6%) associated with the preparation. As determined by histone-H1 phosphorylation after DEAE-cellulose DE-52 chromatography, protein kinase C activity with a marked dependence on Ca2+ and lipid (4-fold increase in their presence) was evident in the secretory-granule fraction. Phosphorylation in vitro of the secretory-granule fraction by endogenous and exogenous protein kinase C revealed a protein of Mr 36,000, which by two-dimensional SDS/polyacrylamide-gel electrophoresis showed multiple sites of phosphorylation. The Mr-36,000 protein was not found in cytosolic or plasma-membrane fractions and was not phosphorylated by the catalytic subunit of cyclic AMP-dependent protein kinase. Several secretory-granule proteins served as substrates for the catalytic subunit, the most prominent of which were of Mr 63,000, 23,000 and 21,000. From these data, we suggest that phosphorylation of secretory-granule-associated proteins by protein kinase C and by cyclic AMP-dependent protein kinase may be important in secretion regulation in the anterior pituitary gland. PMID- 3026316 TI - The decrease in phosphatidylinositol 4,5-bisphosphate in ADP-stimulated washed rabbit platelets is not primarily due to phospholipase C activation. AB - Addition of 10 micron-ADP to washed rabbit platelets caused platelet shape change and aggregation without release of the contents of the amine-storage granules, and caused a transient decrease (8.8% at 10 s) in the amount of phosphatidylinositol 4,5-bisphosphate (PIP2). By 20 s the decrease in PIP2 was no longer apparent, but by 60 s the amount of PIP2 was again decreased. Addition of thrombin (1 unit/ml), which causes platelet shape change, aggregation and the release of the contents of the amine-storage granules, caused a decrease in the amount of PIP2 (8.0% at 10 s); at 60 s the amount of PIP2 was not significantly different from that in controls. In platelets prelabelled with [3H]glycerol, the specific radioactivity of PIP2 was increased at 10 s in ADP-stimulated platelets, and unchanged in thrombin-stimulated platelets. In platelets prelabelled with [3H]inositol and incubated with 20 mM-Li+ to inhibit the degradation of the inositol phosphates to inositol, there was no increase in the labelling of inositol trisphosphate (IP3) upon stimulation with ADP. In contrast, stimulation with thrombin caused a significant increase in the labelling of IP3 at 10 s. These differences in the changes in polyphosphoinositide metabolism in ADP- and thrombin-stimulated platelets are consistent with the hypothesis that the decrease in PIP2 in ADP-stimulated platelets may be due not to degradation of PIP2 by phospholipase C, but rather to a shift in the equilibrium between PIP2 and phosphatidylinositol 4-phosphate (PIP). Increases in the labelling of phosphatidic acid at 10 s and of inositol bisphosphate and inositol phosphate after 20 s are consistent with phospholipase C being stimulated through some other mechanism that leads to the degradation of PIP and phosphatidylinositol; one possibility is that ADP causes an increase in cytoplasmic Ca2+. PMID- 3026315 TI - Uroporphyrin accumulation produced by halogenated biphenyls in chick-embryo hepatocytes. Reversal of the accumulation by piperonyl butoxide. AB - Cultures of chick-embryo hepatocytes were used to study the mechanism by which 3,4,3',4'-tetrachlorobiphenyl and 2,4,5,3',4'-pentabromobiphenyl cause accumulation of uroporphyrin. In a previous paper, an isoenzyme of cytochrome P 450 induced by 3-methylcholanthrene had been implicated in this process [Sinclair, Bement, Bonkovsky & Sinclair (1984) Biochem. J. 222, 737-748]. Cells treated with 3,4,3',4'-tetrachlorobiphenyl and 5-aminolaevulinate accumulated uroporphyrin and heptacarboxyporphyrin, whereas similarly treated cells accumulated protoporphyrin immediately after piperonyl butoxide was added. Piperonyl butoxide also restored haem synthesis as detected by incorporation of radioactive 5-aminolaevulinate into haem, and decrease in drug-induced 5 aminolaevulinate synthase activity. The restoration of synthesis of protoporphyrin and haem by piperonyl butoxide was not affected by addition of cycloheximide, indicating recovery was probably not due to protein synthesis de novo. Piperonyl butoxide also reversed uroporphyrin accumulation caused by 3,4,5,3',4',5'-hexachlorobiphenyl, mixtures of other halogenated biphenyls, lindane, parathion, nifedipine and verapamil. The effect of piperonyl butoxide was probably not due to inhibition of metabolism of these compounds, since the hexachlorobiphenyl was scarcely metabolized. Other methylenedioxyphenyl compounds, as well as ellipticine and acetylaminofluorene, also reversed the uroporphyrin accumulation caused by 3,4,3',4'-tetrachlorobiphenyl. SKF-525A (2 dimethylaminoethyl-2,2-diphenyl valerate) did not reverse the uroporphyrin accumulation caused by the halogenated biphenyls, but did reverse that caused by phenobarbital and propylisopropylacetamide. We conclude that the mechanism of the uroporphyrin accumulation cannot be due to covalent binding of activated metabolites of halogenated compounds to uroporphyrinogen decarboxylase. PMID- 3026317 TI - The cloning and expression of the aroL gene from Escherichia coli K12. Purification and complete amino acid sequence of shikimate kinase II, the aroL gene product. AB - The aroL gene encoding the enzyme shikimate kinase II was cloned from Escherichia coli K12. Construction of over-expressing strains permitted for the first time the purification to homogeneity of a monofunctional shikimate kinase. The complete amino acid sequence of shikimate kinase II was determined by a combined nucleotide and direct amino acid sequencing strategy. E. coli shikimate kinase II is a monomeric enzyme containing 173 amino acid residues with a calculated Mr 18,937. The amino acid sequence contains a region homologous with other kinases and ATP-requiring enzymes. Evidence is presented suggesting that the transcriptional start site of the aroL gene is located within a potential operator site. PMID- 3026318 TI - Inhibition of hepatic gluconeogenesis by the Rp-diastereomer of adenosine cyclic 3',5'-phosphorothioate. AB - The specific intracellular cyclic AMP-dependent protein kinase antagonist, the Rp diastereomer of adenosine cyclic 3',5'-phosphorothioate (Rp-cAMPS), inhibited both basal and cyclic AMP-agonist-induced rates of gluconeogenesis in hepatocytes isolated from fasted rats. Incubation of the cells in the presence of pyruvate and lactate and either the Sp-diastereomer of adenosine cyclic 3',5' phosphorothioate (Sp-cAMPS) or glucagon produced a concentration-dependent increase in the rate of gluconeogenic glucose production which was shifted to higher concentrations of Sp-cAMPS or glucagon in the presence of Rp-cAMPS. Incubation of the cells with Rp-cAMPS in the absence of agonist produced no increase in the rate of glucose production and, in most cases, 100 microM-Rp cAMPS resulted in 14-20% decrease in the substrate-stimulated rate of glucose production. Sp-cAMPS-induced gluconeogenesis was inhibited half-maximally at 1 microM-Rp-cAMPS and glucagon-induced gluconeogenesis was inhibited half-maximally at 12 microM-Rp-cAMPS. Approx. 10-15% of the inhibition of gluconeogenesis observed in the presence of Rp-cAMPS was due to conversion of glucose 6-phosphate to liver glycogen, consistent with Rp-cAMPS-induced reactivation of glycogen synthase. The remaining 85-90% inhibition of gluconeogenic glucose production resulted from the action of Rp-cAMPS on the cyclic AMP-sensitive enzymes controlling the rate of gluconeogenesis. PMID- 3026319 TI - Aromatic hydroxylation as a potential measure of hydroxyl-radical formation in vivo. Identification of hydroxylated derivatives of salicylate in human body fluids. AB - Attack by .OH radicals, generated by a Fenton system, upon salicylate produces 2,3-dihydroxybenzoate and 2,5-dihydroxybenzoate as major products and catechol as a minor product. H.p.l.c. separation combined with electrochemical detection was used to identify and quantify 2,3-dihydroxybenzoate and 2,5-dihydroxybenzoate in human plasma and synovial fluid. We propose that conversion of salicylate into 2,3-dihydroxybenzoate, or of other aromatic compounds into specific hydroxylated products, may be a useful assay for .OH formation in the human body. PMID- 3026321 TI - Effects of adrenalectomy on binding to and actions of adrenergic receptors. AB - Adrenalectomy results in significant changes in the mechanism of adrenergic activation of hepatic glycogenolysis. In adrenalectomized rats a greater role for the beta-adrenergic receptor is observed, whereas the alpha 1-adrenergic-mediated phosphorylase activation declines. Our present findings document that adrenalectomy causes a significant decrease in the high-affinity population of the alpha 1-adrenergic receptor labelled with [3H]adrenaline. Our data indicate a large increase in the number of beta-adrenergic binding sites after adrenalectomy. This increase was not consistent with the observed modest increase in the beta-adrenergic-mediated activation of cyclic AMP accumulation and glycogen phosphorylase. When alpha-adrenergic antagonists are present along with the catecholamine, a 100% increase in the adrenaline-mediated accumulation of cyclic AMP in hepatocytes from adrenalectomized rats was observed. Adrenalectomy was also shown to cause a significant increase in the hepatic alpha 2-adrenergic binding sites. These data are consistent with an inhibitory role on the beta adrenergic-mediated activation of glycogenolysis by the hepatic alpha 2 adrenergic receptor in adrenalectomy. PMID- 3026320 TI - Free-radical chain oxidation of 2-nitropropane initiated and propagated by superoxide. AB - The superoxide radical O2.-, whether produced by the xanthine/xanthine oxidase reaction or infused as KO2, solubilized by a crown ether in dry dimethyl sulphoxide, initiated a free-radical chain oxidation of anionic 2-nitropropane. Superoxide dismutase, but not catalase, inhibited oxidation of the nitroalkane. Xanthine oxidase suffered a syncatalytic inactivation, during the co-oxidation of 2-nitropropane, which was reversed by dialysis. Cyanide exacerbated this syncatalytic inactivation and rendered it irreversible. The frequently observed oxidations of nitroalkanes by flavoenzymes now need to be re-examined to clarify the extent to which O2.--initiated free-radical chain oxidation contributed to the overall nitroalkane oxidation. PMID- 3026323 TI - Molecular forms of myeloperoxidase in human plasma. AB - A radioimmunoassay for myeloperoxidase was established with the use of affinity purified anti-(human myeloperoxidase) immunoglobulins. By the use of ion-exchange followed by immunoaffinity chromatography a preparation of immunoreactive, catalytically active myeloperoxidase was obtained from fresh human plasma. In non denaturing gel electrophoresis, the plasma preparation showed about four catalytically active components of mobility very similar to that of the granulocyte enzyme. SDS/polyacrylamide-gel electrophoresis combined with protein blotting showed that the two polypeptides of strongest antigenicity in the plasma preparation corresponded in Mr to the large and the small subunits of the granulocyte enzyme. In addition, the plasma preparation contained a higher-Mr immunoreactive polypeptide, possibly a precursor form of the enzyme, together with another of Mr similar to that of the large subunit of eosinophil peroxidase. PMID- 3026322 TI - NADPH oxidase of guinea-pig macrophages catalyses the reduction of ubiquinone-1 under anaerobic conditions. AB - The stimulation-specific NADPH-dependent reduction of ubiquinone-1 (Q-1) in guinea-pig macrophages was studied. The activity was due neither to any modified product of the phagocytosis-specific NADPH oxidase nor to non-specific diaphorases of the cells, since the activity was measured in sonicated or detergent-disrupted cells by subtracting the activity in the resting cells from that in cells activated by phorbol 12-myristate 13-acetate. The activity was not mediated by superoxide anions, since strict anaerobic conditions were employed. The anaerobic reduction of Q-1 was NADPH-specific, like superoxide formation under aerobic conditions, and its maximal velocity was also essentially the same as that of superoxide formation. The oxidase does not directly reduce Q-1 under aerobic conditions [Nakamura, Murakami, Umei & Minakami (1985) FEBS Lett. 186, 215-218], and the electron transfer from NADPH to cytochrome c by the oxidase under aerobic conditions was not enhanced by the addition of Q-1. The observations indicate that the phagocytosis-specific NADPH oxidase reduces Q-1 and that oxygen competes with the reduction of Q-1. Q-1 seems to accept electrons not from the intermediary electron carriers of the oxidase but from the terminal oxygen-reducing site of the enzyme. PMID- 3026325 TI - Excretion of laccase by sycamore (Acer pseudoplatanus L.) cells. Effects of a copper deficiency. AB - Copper-deprived sycamore (Acer pseudoplatanus) cells do not excrete molecules of active laccase in their culture medium. In the range of 2-100 micrograms of copper initially present per litre of nutrient solution, the total laccase activity measured in the cell suspensions at the end of the exponential phase of growth was closely proportional to the amount of added copper. However, copper deprived cells excreted the laccase apoprotein (laccase without copper) at the same rate as copper-supplied cells excreted the active, copper-containing, laccase. When the culture medium was initially supplied with limiting amounts of copper, the active laccase was excreted until all copper molecules were metabolized. Thereafter, the laccase apoprotein was excreted. Consequently, at the end of the exponential phase of growth, the cell supernatants contained a mixture of apoprotein and copper-containing laccase. After purification and concentration, this mixture of copper-containing laccase (blue) and laccase apoprotein (slightly yellow) showed a yellow-green colour. Under copper-limiting culture conditions an equivalent decrease of Type 1, Type 2 and Type 3 Cu2+ was observed. Addition of copper to copper-deficient enzyme solutions does not result in a recovery of the enzyme activity. However, when added to copper-deficient sycamore-cell suspensions, copper induced a recovery of the excretion of active enzyme, at a normal rate, within about 10 h. The first molecules of active laccase were excreted after 3-4 h. PMID- 3026324 TI - Kinetic studies of the reduction of neutrophil cytochrome b-558 by dithionite. AB - The reduction with dithionite of neutrophil cytochrome b-558, implicated in superoxide generation by activated neutrophils, was investigated by a stopped flow technique in non-ionic-detergent extracts of the membranes and in crude membrane particles. The dependence of the pseudo-first-order rate constants on the concentration of dithionite was consistent with a mechanism of reduction that involves the dithionite anion monomer SO2.- as the reactive species. The estimated second-order rate constant was 7.8 X 10(6) M-1 X S-1 for Lubrol PX solubilized cytochrome b-558 and 5.1 X 10(6) M-1 X S-1 for the membrane-bound protein. The similarity of the kinetic constants suggests that solubilization did not introduce gross changes in the reactive site. Imidazole and p chloromercuribenzoate, known as inhibitors of NADPH oxidase, did not affect significantly cytochrome b-558 reduction rates. The reaction rate of cytochrome b 558 with dithionite exhibited a near-zero activation energy. The first-order rate constant for reduction decreased with increasing ionic strength, indicating a positive effective charge on the reacting protein. PMID- 3026326 TI - Induction of haem synthesis in Hep G2 human hepatoma cells by dimethyl sulphoxide. A transcriptionally activated event. AB - Exposure of cultured human hepatoma cells (Hep G2) to medium containing 2% (v/v) dimethyl sulphoxide resulted in an approximate doubling in the activity of delta aminolaevulinate dehydratase, an increase in the haem content and a decreased growth rate; induced enzyme activity was decrease by 50% after treatment with alpha-amanitin. The findings are strikingly similar to those seen in murine Friend-virus-transformed erythroleukaemia cells. PMID- 3026328 TI - Electron nuclear double-resonance (ENDOR) spectroscopy of amine oxidase from pig plasma. AB - Electron nuclear double-resonance ('ENDOR') spectroscopic studies on pig plasma amine oxidase have been carried out at 15 K. Deuterium-exchange studies show the presence of two sets of exchangeable protons, probably from two water molecules; from the magnitude of their hyperfine couplings, one is assigned to be equatorially, and the other axially, co-ordinated. Only one 14N hyperfine coupling is observed, suggesting that the bonding of all amino acid (histidine) or organic cofactor ligands is similar. Upon addition of azide, a further hyperfine coupling to nitrogen is observed which is smaller than that observed for the native enzyme; the hyperfine couplings to the remaining nitrogens are slightly altered. PMID- 3026327 TI - Regulation of superoxide generation by myeloperoxidase during the respiratory burst of human neutrophils. AB - The role of myeloperoxidase in the regulation of the respiratory burst of human neutrophils activated by the chemotactic peptide (N-formyl-L-methionyl-L-leucyl-L phenylalanine) plus cytochalasin B was determined by using anti-(human myeloperoxidase) antibody. The respiratory burst activated under these conditions consisted of an initial (1-2 min) phase with high rates of O2 uptake, luminol dependent chemiluminescence and superoxide radical (O2-.) generation and a second, more sustained, phase of lower magnitude of chemiluminescence and O2 uptake: O2-. generation did not occur during this second phase. In cell suspensions stimulated in the presence of anti-(human myeloperoxidase) antibody, the magnitude of the initial phase of both O2 uptake and O2-. generation was unaffected, but these high rates were maintained over much longer periods than in control suspensions. It is therefore proposed that a product of myeloperoxidase normally regulates the duration of O2-. generation during the respiratory burst, possibly by inhibition of NADPH oxidase. PMID- 3026329 TI - Characterization of a retinylmonophosphatase in the plasma membrane of mouse brain. AB - Retinylmonophosphatase (RMPase) activity in mouse brain paralleled the subcellular distribution of the plasma-membrane marker Na+ + K+-dependent ATPase. The enzyme had a pH optimum between 5.5 and 7.0. The enzyme demonstrated linear kinetics with respect to time and both protein and substrate concentrations. RMPase was saturated by low retinyl monophosphate (RMP) concentrations and exhibited an apparent Km of 4.6 microM. The enzyme did not require MgCl2 for activity, and in fact assays were routinely run in the presence of 10 mM-Na2EDTA. In general, detergents inhibited the enzyme, with 0.05% Triton X-100 causing a 30% loss of activity. Phosphatidic acid was also inhibitory, but phosphatidylcholine and sphingomyelin stimulated phosphatase activity. RMPase was inhibited 35% by 5 mM concentrations of fluoride, phosphate or pyrophosphate. A series of other phosphorylated compounds, including glucose 6-phosphate, alpha glycerophosphate, ATP, AMP, p-nitrophenyl phosphate and thiamin pyrophosphate, showed little or no inhibition. RMPase activity differed in several characteristics from that previously reported for dolichylmonophosphatase. It is concluded that RMP could play a distinct role in the plasma membrane. PMID- 3026330 TI - Evidence for two GTPases activated by thrombin in membranes of human platelets. AB - Thrombin inhibits adenylate cyclase and stimulates GTP hydrolysis by high affinity GTPase(s) in membranes of human platelets at almost identical concentrations. Both of these thrombin actions are similar to those observed with agonist-activated alpha 2-adrenoceptors coupling to the inhibitory guanine nucleotide-binding protein N1. However, stimulation of GTP hydrolysis caused by adrenaline (alpha 2-adrenoceptor agonist) and by thrombin at maximally effective concentrations was partially additive, whereas with regard to adenylate cyclase inhibition no additive response was observed. Furthermore, treatment of platelet membranes with pertussis toxin, which inactivates Ni and largely abolishes thrombin- and adrenaline-induced adenylate cyclase inhibition and adrenaline induced GTPase stimulation, decreased the thrombin-induced stimulation of GTP hydrolysis by only about 30%. Additionally, the thiol reagent N-ethylmalemide (NEM) at rather low concentrations abolished thrombin- and adrenaline-induced stimulation of GTP hydrolysis was decreased by only 30-40% by treatment of platelet membranes with even high concentrations of NEM. Treatment with cholera toxin, which inhibits GTPase activity of the Ns (stimulatory guanine nucleotide binding) protein, has no effect on thrombin-stimulated GTP hydrolysis. The data suggest that thrombin interaction with its receptor sites in platelet membranes leads to stimulation of two GTP-hydrolysing enzymes. One of these enzymes is apparently Ni and is also activated by agonist-activated alpha 2-adrenoceptors and is inactivated by pertussis toxin and NEM treatment. The other GTP hydrolysing enzyme activated by thrombin may represent a guanine nucleotide binding protein apparently involved in the coupling of thrombin receptors to the phosphoinositide phosphodiesterase. PMID- 3026331 TI - Polyphosphoinositide metabolism in baby-hamster kidney cells infected with herpes simplex virus type 1. AB - The incorporation of [32P]Pi and [3H]inositol into the inositol lipids of baby hamster kidney cells was studied in herpes-simplex-virus-type-1(HSV-1)-infected and mock-infected cells. The infection was conducted during incorporation of, as well as after prelabelling with, the precursors. These methods were used in order to study both synthesis de novo of, and steady-state changes in, the phosphoinositides. Both with infection during labelling, and after prelabelling, we found increased [32P]- and [3H]-phosphatidylinositol 4,5-bisphosphate (PIP2) and decreased [32P]- and [3H]-phosphatidylinositol 4-monophosphate in infected as compared with mock-infected cells, whereas no effect was observed on phosphatidylinositol. This altered inositol-lipid metabolism was (at least in the case of PIP2) not present until 3-6 h after infection and remained stable, or increased slightly, throughout the infection period. Polyphosphoinositide metabolism constitutes an important step in signal processing in many forms of cellular stimulation, and the results obtained suggest that HSV-1 infection may induce such events in our cell system. PMID- 3026332 TI - Stimulation of glycogenolysis by adenine nucleotides in the perfused rat liver. AB - Infusion of adenine nucleotides and adenosine into perfused rat livers resulted in stimulation of hepatic glycogenolysis, transient increases in the effluent perfusate [3-hydroxybutyrate]/[acetoacetate] ratio, and increased portal vein pressure. In livers perfused with buffer containing 50 microM-Ca2+, transient efflux of Ca2+ was seen on stimulation of the liver with adenine nucleotides or adenosine. ADP was the most potent of the nucleotides, stimulating glucose output at concentrations as low as 0.15 microM, with half-maximal stimulation at approx. 1 microM, and ATP was slightly less potent, half-maximal stimulation requiring 4 microM-ATP. AMP and adenosine were much less effective, doses giving half-maximal stimulation being 40 and 20 microM respectively. Non-hydrolysed ATP analogues were much less effective than ATP in promoting changes in hepatic metabolism. ITP, GTP and GDP caused similar changes in hepatic metabolism to ATP, but were 10 20 times less potent than ATP. In livers perfused at low (7 microM) Ca2+, infusion of phenylephrine before ATP desensitized hepatic responses to ATP. Repeated infusions of ATP in such low-Ca2+-perfused livers caused homologous desensitization of ATP responses, and also desensitized subsequent Ca2+-dependent responses to phenylephrine. A short infusion of Ca2+ (1.25 mM) after phenylephrine infusion restored subsequent responses to ATP, indicating that, during perfusion with buffer containing 7 microM-Ca2+, ATP and phenylephrine deplete the same pool of intracellular Ca2+, which can be rapidly replenished in the presence of extracellular Ca2+. Measurement of cyclic AMP in freeze-clamped liver tissue demonstrated that adenosine (150 microM) significantly increased hepatic cyclic AMP, whereas ATP (15 microM) was without effect. It is concluded that ATP and ADP stimulate hepatic glycogenolysis via P2-purinergic receptors, through a Ca2+-dependent mechanism similar to that in alpha-adrenergic stimulation of hepatic tissue. However, adenosine stimulates glycogenolysis via P1-purinoreceptors and/or uptake into the cell, at least partially through a mechanism involving increase in cyclic AMP. Further, the hepatic response to adenine nucleotides may be significant in regulating hepatic glucose output in physiological and pathophysiological states. PMID- 3026333 TI - Stimulation of phosphoinositide hydrolysis by oxytocin and the mechanism by which oxytocin controls prostaglandin synthesis in the ovine endometrium. AB - Slices of caruncular endometrium from steroid-treated ovariectomized sheep were incubated with myo-[2-3H]inositol to label tissue phosphatidylinositol. Effects of oxytocin were determined on the rate of incorporation of radioactivity into phosphatidylinositol and on the hydrolysis of phosphoinositides to inositol phosphates and diacylglycerol. Incorporation of radioactivity into phosphatidylinositol was linear during 2 h incubations; 10(-7) M (100 nM) oxytocin caused a 2.8-fold increase in the rate of incorporation. In the presence of Li+, addition of 10(-7) M-oxytocin to slices in which phosphatidylinositol was pre-labelled caused mean increase of 40-fold in the incorporation of radioactivity into inositol mono-, bis- and tris-phosphates. Inositol 1,3,4 trisphosphate was quantitatively the major trisphosphate formed. The action of oxytocin on phosphoinositide hydrolysis was dose- and time-dependent, occurring at concentrations within the range observed in plasma during episodes of secretion in vivo, and with a time course comparable with that of the action of oxytocin on uterine prostaglandin production. The effect of oxytocin on incorporation of radioactivity into inositol phosphates was not affected by inhibitors of prostaglandin synthesis. Diacylglycerol 1- and 2-lipases in caruncular endometrium converted up to 72% of added 2 [3H]arachidonyldiacylglycerol into [3H]arachidonic acid during 30 min incubations at pH 7.0. Caruncular endometrium contained 1.49 mumol of phosphatidylinositol/g, representing approx. 0.2 mumol/g of phosphatidylinositol arachidonic acid. It is proposed that the stimulation of endometrial prostaglandin synthesis by oxytocin is accounted for by increased hydrolysis of phosphoinositides to diacylglycerol and inositol phosphates with subsequent release of arachidonic acid from diacylglycerol. PMID- 3026334 TI - Proactivator-dependent activation of procollagenase induced by treatment with EGTA. AB - A new mechanism for activation of the proactivator of procollagenase [Vater, Nagase & Harris (1983) J. Biol. Chem. 258, 9374-9382] has been found. Collagenolytic and other proteolytic enzyme activities in the medium of cultured rabbit synovial fibroblasts were found to be activated by a new mechanism: short term incubation at 37 degrees C performed in the presence of EGTA followed by replacement of Ca2+ during enzyme assay. The crucial event in procollagenase activation is the production of a functional activator enzyme. Activation of procollagenase in the culture medium did not occur when proactivator was removed by immunoprecipitation. Proteolytic activity of proactivator was fully activated, whereas procollagenase alone could not be activated by the same sequence. EGTA treatment of the culture medium at 0 degrees C did not result in enzyme activation if Ca2+ was replaced before incubation at 37 degrees C. Certain other bivalent metal ions (e.g. Sn2+, Cd2+, Zn2+ and Mn2+) could substitute for Ca2+ to stabilize the proactivator as a zymogen and therefore prevent the appearance of proteolytic activity in culture medium. Isolation of proactivator and procollagenase from EGTA-treated radiolabelled culture medium by immunoprecipitation and subsequent analyses by fluorography revealed that a time dependent proteolysis of both molecules occurred after replacement of Ca2+ and incubation at 37 degrees C. However, comparison of enzyme activity with fluorographic analyses showed that the maximal activation of both enzymes was achieved before any detectable decrease in Mr. The results suggest that the activation of proactivator and the subsequent activation of procollagenase may be initiated by conformational changes in structure of the proactivator molecule produced by removal of stabilizing bivalent metal ions. PMID- 3026335 TI - Oxidation of nicotinamide coenzyme dimers by one-electron-accepting proteins. AB - The nicotinamide nucleotide dimers (NAD)2 and (NADP)2, obtained by electrochemical reduction of NAD+ and NADP+, are able to reduce such single electron acceptors as the proteins cytochrome c, azurin and methaemoglobin, though at different rates. Under the same conditions the reduced nicotinamide coenzymes NADH and NADPH are not able to reduce these proteins at measurable rates unless a catalyst (phenazine methosulphate or NADH-cytochrome c reductase in the case of cytochrome) is present. The redox mechanism seems to involve the formation of an NAD(P). radical that in the presence of O2 gives rise to superoxide (O2.-), since superoxide dismutase inhibited these reactions. PMID- 3026336 TI - Sensitivity to NN'-dicyclohexylcarbodi-imide of proton translocation by mitochondrial NADH:ubiquinone oxidoreductase. AB - Proton extrusion during ferricyanide reduction by NADH-generating substrates or succinate was studied in isolated rat liver mitochondria with the use of optical indicators. NN'-Dicyclohexylcarbodi-imide (DCCD) caused a decrease of 84% in the H+/e- ratio of NADH:cytochrome c reduction, but a decrease of only 49% in that of succinate:cytochrome c reduction, even though electron transfer was decreased equally in both spans. The data indicate that a DCCD-sensitive channel operates in the NADH:ubiquinone oxidoreductase region of the respiratory chain. PMID- 3026337 TI - Age and hormonal dependence of tonin levels in rat submandibular gland as determined by a new direct radioimmunoassay. AB - A simple and sensitive direct radioimmunoassay for tonin (EC.3.4.99.-) has been developed. This assay incorporates a modified and convenient poly(ethylene glycol) technique for separation of free from bound tonin. A rabbit antiserum in a final dilution of 1:160,000 was used and the purified tonin was labelled with 125I by using a lactoperoxidase method. It detects 20 pg of immunoreactive tonin per tube. Serial dilutions of rat submandibular gland extracts showed complete parallelism with tonin standard curves. No cross-reactivity with rat tissue kallikrein was seen. Intra- and inter-assay errors were 3.2 and 5.6%, respectively. Using this assay, immunoreactive tonin was detected in the rat submandibular gland as early as 3 weeks after birth (body wt. approximately 50-60 g). Tonin levels are shown to be dependent on age and sex with significantly higher levels in male than in female rats. Castration results in decrease of tonin levels and 17 alpha-methyltestosterone replacement reversed the level to higher than the sham-operated control rats. Cortisol treatment increased, but thyroxine or oestradiol had no effect, on tonin levels in the submandibular gland of castrated rats. This newly developed radioimmunoassay can now be used to measure low levels of tonin in various tissues and body fluids to address questions about its regulation and functional significance. PMID- 3026338 TI - The effect of phospholipids on the activation of protein C by the human thrombin thrombomodulin complex. AB - Human thrombomodulin, an endothelial-cell-membrane glycoprotein, has been purified from placenta by Triton X-100 extraction and by affinity chromatography on concanavalin A-Sepharose and thrombin-Sepharose. It has been characterized by its ability to promote the activation of human protein C by human alpha-thrombin in the presence of Ca2+ and fulfilled the requirements of a cofactor. Reconstitution of thrombomodulin into phospholipid vesicles containing anionic phospholipids resulted in an increased rate of activation of protein C. Cardiolipin and vesicles containing phosphatidylcholine/phosphatidylserine (1:1, w/w) were the most effective. The apparent Km of the thrombin-thrombomodulin complex for protein C was 2 microM. It was not changed in the presence of phospholipid, whereas the Vmax. could be apparently increased up to 3.2-fold depending on the phospholipid and on its concentration, the catalytic-centre activity reaching 15.7 mol of activated protein C formed/min per mol of thrombin. Above their optimal concentrations, phospholipids inhibited the amidolytic activity of activated protein C. Phospholipids had no effect on the activation of 4-carboxyglutamic acid-domainless protein C, a proteolytic derivative of protein C lacking the 4-carboxyglutamic acid residues. These results show that the positive effect of anionic phospholipids in the activation of protein C by the thrombin-thrombomodulin complex involves a Ca2+-dependent interaction between protein C and phospholipids. They suggest that the enhancement of thrombomodulin activity by such phospholipids may be of functional significance. PMID- 3026339 TI - Differential effects of chlorpromazine on secretion, protein phosphorylation and phosphoinositide metabolism in stimulated platelets. AB - Increasing concentrations of chlorpromazine (30-500 microM) caused a progressive lysis of gel-filtered platelets, as monitored by the extracellular appearance of cytoplasmic ([14C]adenine-labelled) adenine nucleotides. The chlorpromazine induced lysis was markedly enhanced by thrombin and phorbol ester, and complete cytolysis was found at chlorpromazine concentrations of 100 microM and above in the presence of thrombin. At non-lytic concentrations, chlorpromazine caused a dramatic increase in the thrombin- or phorbol ester-mediated incorporation of 32P into phosphatidylinositol 4-phosphate and, to a lesser extent, into phosphatidylinositol 4,5-bisphosphate in platelets pulse-labelled with [32P]Pi. Chlorpromazine alone also caused an incorporation of 32P into the phosphoinositides. Non-lytic concentrations of chlorpromazine had no effect on the phosphorylation of the 47 kDa protein (regarded as the substrate for protein kinase C), but markedly inhibited the accompanying secretion of ATP + ADP and beta-hexosaminidase when platelets were incubated with 0.17 microM-phorbol ester or 0.1-0.2 unit of thrombin/ml. At lower concentrations of thrombin, chlorpromazine did not inhibit, but slightly enhanced, secretion. A protein of 82 kDa was phosphorylated during the interaction of platelets with thrombin and phorbol ester, and this phosphorylation was enhanced by chlorpromazine (non lytic). These results suggest that the previously reported inhibition of protein kinase C by chlorpromazine is probably non-specific and due to cytolysis. However, since non-lytic concentrations of chlorpromazine inhibit secretion, but not protein kinase C, in platelets, activation of protein kinase C is not involved in the stimulation-secretion coupling, or chlorpromazine acts at a step after kinase activation. Possible mechanisms of this inhibition by chlorpromazine are discussed in the light of its effect on phosphoinositide metabolism and protein phosphorylation. PMID- 3026340 TI - Comparisons of copper deficiency states in the murine mutants blotchy and brindled. Changes in copper-dependent enzyme activity in 13-day-old mice. AB - The activity of two copper-dependent enzymes, cytochrome c oxidase and copper, zinc-superoxide dismutase, was determined in six tissues of age-matched (13-day old) copper-deficient mutant and normal mice. In the two mutants 'brindled' and 'blotchy', brain, heart and skeletal muscle had significant enzyme deficiencies. Cytochrome c oxidase was more severely affected than was superoxide dismutase. In these three tissues the degree of deficiency could be correlated with decreased copper concentration; however, enzyme activity was normal in liver, kidney and lung, despite abnormal copper concentrations in these tissues. In nutritionally copper-deficient mice, all six tissues showed decreased enzyme activity, which was most marked in brain, heart and skeletal muscle, the tissues which showed enzyme deficiencies in the mutants. Analysis in vitro of cytochrome c oxidase (temperature coefficient = 2) at a single temperature was found to underestimate the deficiency of this enzyme in hypothermic copper-deficient animals. Cytochrome c oxidase deficiency may therefore be sufficiently severe in vivo to account for the clinical manifestations of copper deficiency. An injection of copper (50 micrograms of Cu+) at 7 days increased cytochrome c oxidase activity by 13 days in all deficient tissues of brindled mice, and in brain and heart from blotchy mice. However, skeletal-muscle cytochrome c oxidase in blotchy mutants did not respond to copper injection. Cytochrome c oxidase activity increased to normal in all tissues of nutritionally copper-deficient mice after copper injection, except in the liver. Hepatic enzyme activity remained severely deficient despite a liver copper concentration three times that found in copper-replete controls. Superoxide dismutase activity did not increase with treatment in either mutant, but its activity was higher than control levels in nutritionally deficient mice after injection. This difference is probably due to sequestration of copper in mutant tissue such as kidney, but a defect in the copper transport pathway to superoxide dismutase cannot be excluded. PMID- 3026341 TI - Antagonistic regulation of the glucose/glucose 6-phosphate cycle by insulin and glucagon in cultured hepatocytes. AB - Flux through the glucose/glucose 6-phosphate cycle in cultured hepatocytes was measured with radiochemical techniques. Utilization of [2-3H]glucose was taken as a measure of glucokinase flux. Liberation of [14C]glucose from [U-14C]glycogen and from [U-14C]lactate, as well as the difference between the utilization of [2 3H]glucose and of [U-14C]glucose, were taken as measures of glucose-6-phosphatase flux. At constant 5 mM-glucose and 2 mM-lactate concentrations insulin increased glucokinase flux by 35%; it decreased glucose-6-phosphatase flux from glycogen by 50%, from lactate by 15% and reverse flux from external glucose by 65%, i.e. overall by 40%. Glucagon had essentially no effect on glucokinase flux; it enhanced glucose-6-phosphatase flux from glycogen by 700%, from lactate by 45% and reverse flux from external glucose by 20%, i.e. overall by 110%. At constant glucose concentrations cellular glucose 6-phosphate concentrations were essentially not altered by insulin, but were increased by glucagon by 230%. In conclusion, under basic conditions without added hormones the glucose/glucose 6 phosphate cycle showed only a minor net glucose uptake, of 0.03 mumol/min per g of hepatocytes; this flux was increased by insulin to a net glucose uptake of 0.21 mumol/min per g and reversed by glucagon to a net glucose release of 0.22 mumol/min per g. Since the glucose 6-phosphate concentrations after hormone treatment did not correlate with the glucose-6-phosphatase flux, it is suggested that the hormones influenced the enzyme activity directly. PMID- 3026342 TI - Further characterization of the platinum-reactive component of the alpha 2 macroglobulin-receptor recognition site. AB - alpha 2-Macroglobulin (alpha 2M)-methylamine that had been allowed to react with cis-dichlorodiammineplatinum(II) (cis-DDP) bound with greatly reduced affinity to specific alpha 2M receptors, as determined by macrophage binding studies in vitro and plasma-clearance experiments in vivo. Subsequent reaction with diethyl dithiocarbamate completely restored receptor recognition function. The optimal effect was obtained when the diethyl dithiocarbamate concentration was twice the total platinum concentration. alpha 2M-methylamine that was allowed to react with H2O2 competed less effectively for specific cell-surface binding sites, as demonstrated by studies both in vivo and in vitro. The apparent dissociation constant was increased nearly 7-fold by a 15 min exposure to H2O2. alpha 2M methylamine was affected significantly less by the H2O2 exposure after pretreatment with cis-DDP. Amino acid analysis indicated that H2O2 treatment of alpha 2M modified 19 of the 25 methionine residues per alpha 2M subunit. Pretreatment with cis-DDP protected two to four of these methionine residues. The only other residue altered by H2O2 treatment of alpha 2M was histidine. A net decrease of two histidine residues per subunit was observed, but cis-DDP pretreatment did not alter this result. In order to rule out the slight possibility that histidine modification might account for the observed H2O2 induced loss in receptor recognition, diethyl pyrocarbonate was employed as a histidine-modifying reagent. This treatment modified 53 histidine residues in both native and fast-form alpha 2M. Fast-form alpha 2M was still recognized by the alpha 2M receptor, as determined by studies both in vivo and in vitro; however, a fraction of the modified protein now cleared via the acyl-low-density lipoprotein receptor as well. Reaction of diethyl pyrocarbonate-treated alpha 2M with hydroxylamine reversed derivatization of 43 of the 53 histidine residues. Moreover, this treatment also resulted in an alpha 2M fast-form preparation that was recognized only by the alpha 2M receptor. It is concluded that cis-DDP and H2O2 modify a critical methionine residue in the primary sequence of the alpha 2M receptor recognition site. PMID- 3026343 TI - Isolation and properties of the glycolytic enzymes from Zymomonas mobilis. The five enzymes from glyceraldehyde-3-phosphate dehydrogenase through to pyruvate kinase. AB - The five glycolytic enzymes glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate kinase, phosphoglycerate mutase, enolase and pyruvate kinase were each purified from extracts of Zymomonas mobilis cells, by using dye-ligand chromatography as the principal step. Two procedures, producing three and two of the enzymes respectively, are described in detail. Z. mobilis glyceraldehyde phosphate dehydrogenase was found to be similar in most respects to the enzyme from other sources, except for having a slightly larger subunit size. Phosphoglycerate kinase has properties typical for this enzyme; however, it did not show the sulphate activation effects characteristic of this enzyme from most other sources. Phosphoglycerate mutase is a dimer, partially independent of 2,3 bisphosphoglycerate, and has a high specific activity. Enolase was found to be octameric; otherwise its properties were very similar to those of the yeast enzyme. Pyruvate kinase is unusual in being dimeric, and not requiring K+ for activity. It is not allosterically activated by sugar phosphates, having a high activity in the absence of any effectors. Some quantitative differences in the relative amounts of these enzymes, compared with eukaryotic species, are ascribed to the fact that Z. mobilis utilizes the Entner-Doudoroff pathway rather than the more common Embden-Meyerhoff glycolytic route. PMID- 3026345 TI - An e.p.r. study of the non-equivalence of the copper sites of caeruloplasmin. AB - The two Type 1 (blue) copper-binding sites of caeruloplasmin were spectroscopically differentiated by the kinetic analysis of the e.p.r. spectra during the redox cycle. One blue copper, with a hyperfine splitting constant (A parallel) of 6.8 mT, which was rapidly reduced, was not reoxidized by oxygen, whereas it was reoxidized by H2O2. The other blue copper (A parallel = 5.8 mT), which was reduced slowly, was rapidly reoxidized by either oxygen or H2O2. A conformational change of the Type 2 copper was concomitant with the fast reduction of Type 1 copper, whereas its reduction occurred during the slow phase. This sequence of events was reversed in the reoxidation step, that is, the Type 2 copper reappeared rapidly as the species with altered conformation and reverted to the symmetry typical of the native state in the slow phase. The specific reaction of a blue-copper site with the H2O2 can tentatively be related to the established ability of caeruloplasmin to prevent 'oxidative' attack of proteins and lipids. PMID- 3026344 TI - Role of a pertussis toxin substrate in the control of lectin-induced cap formation in human neutrophils. AB - We have examined the role of GTP-binding proteins and the associated cyclic AMP- and calcium-related transduction mechanisms in the regulation of capping in human neutrophils. Pertussis toxin (PT), a probe for the GTP-binding protein Ni, abolished capping induced by fluorescein isothiocyanate-conjugated concanavalin A (Con-A), whereas cholera toxin, a probe for the GTP-binding protein Ns, was without effect. Consistent with the latter finding, ligands acting at receptors associated with the Ns protein, namely the prostaglandin E1 and beta-adrenergic agonists, were without effect on the capping reaction. The possible role of mobilization of internal calcium was evaluated by using Quin2-loaded cells. Calcium mobilization was observed at concentrations of Con-A which yielded optimal capping (10 micrograms/ml). Treatment with PT, phorbol myristrate acetate or 8-(NN-diethylamino)octyl-3,4,5-trimethoxybenzoate (TMB-8) abolished both calcium mobilization and capping. Colchicine, which substantially enhanced capping, had no effect on calcium mobilization. At concentrations of the lectin above those required for capping, superoxide generation and enzyme release were noted. These reactions were less susceptible to inhibition by PT, effects being observed only on the Kact. for Con-A-mediated superoxide generation with little effect on the Vmax. The degree of PT-mediated inhibition for enzyme release with Con-A was much lower than that observed with fMet-Leu-Phe. Our results imply that a step involving Ni-mediated calcium mobilization, sensitive to phorbol myristate acetate, is essential to the regulation of capping; a distinct mechanism may be involved in colchicine-mediated enhancement of capping; and Ni may play a relative minor role in the regulation of lectin-mediated exocytosis. PMID- 3026346 TI - Studies on the mechanism of sheep liver cytosolic aldehyde dehydrogenase. The effect of pH on the aldehyde binding reactions and a re-examination of the problem of the site of proton release in the mechanism. AB - Initial-rate measurements and stopped-flow spectrophotometric experiments over a wide range of pH implicate an enzyme group of pKa approximately 6.6 affecting the aldehyde binding reactions. It is possible, though not proved, that the group involved is the cysteine residue involved in catalysis. Stopped-flow fluorescence studies show that a group of pKa greater than 8.5 facilitates hydrolysis of the NADH-containing acyl-enzyme species. The identity of this group is quite unknown. Studies with 4-nitrobenzaldehyde show that this substrate gives marked substrate inhibition at quite low (less than 20 microM) concentrations. The mechanism of catalysis seems to be the same as for propionaldehyde oxidation. It is argued that proton release occurs with both substrates on hydrolysis of the NADH containing acyl-enzyme and not before hydride transfer, as has been previously suggested [Bennett, Buckley & Blackwell (1982) Biochemistry 21, 4407-4413]. PMID- 3026347 TI - Sensitivity of pyruvate dehydrogenase phosphate phosphatase to magnesium ions. Similar effects of spermine and insulin. AB - The effects of Mg2+ on the activity of pyruvate dehydrogenase phosphate phosphatase within intact mitochondria prepared from control and insulin-treated rat epididymal adipose tissue was explored by incubating the mitochondria in medium containing the ionophore A23187. The apparent Ka for Mg2+ was approximately halved in the mitochondria derived from insulin-treated tissue in both the absence and the presence of Ca2+. In this system, the major effect of Ca2+ was also to decrease the apparent Ka for Mg2+, rather than to change the Vmax. of the phosphatase. Damuni, Humphreys & Reed [(1984) Biochem. Biophys. Res. Commun. 124, 95-99] have reported that spermine activates ox kidney pyruvate dehydrogenase phosphate phosphatase. Studies were carried out on phosphatase from pig heart and rat epididymal adipose tissue which confirm and extend this observation. The major effect of spermine is shown to be a decrease in the Ka for Mg2+, which is apparent in both the presence and the absence of Ca2+. Spermine did not affect the sensitivity of the phosphatase to Ca2+ at saturating concentrations of Mg2+. Other polyamines tested were not as effective as spermine. No alteration in the maximum activity or Mg2+-sensitivity of pyruvate dehydrogenase phosphate phosphatase was apparent in extracts of mitochondria from insulin-treated tissue. The close similarity of the effects of spermine and the changes in kinetic properties of pyruvate dehydrogenase phosphate phosphatase within mitochondria from insulin-treated adipose tissue suggests that insulin may activate pyruvate dehydrogenase by increasing the concentration of spermine within the mitochondria. However, it is concluded that insulin is more likely to alter the interaction of the pyruvate dehydrogenase system with some other polybasic intramitochondrial component whose action can be mimicked by spermine. PMID- 3026348 TI - Use of toluene-permeabilized mitochondria to study the regulation of adipose tissue pyruvate dehydrogenase in situ. Further evidence that insulin acts through stimulation of pyruvate dehydrogenase phosphate phosphatase. AB - Rat epididymal-adipose-tissue mitochondria were made selectively permeable to small molecules without the loss of matrix enzymes by treating the mitochondria with toluene under controlled conditions. With this preparation the entire pyruvate dehydrogenase system was shown to be retained within the mitochondrial matrix and to retain its normal catalytic activity. By using dilute suspensions of these permeabilized mitochondria maintained in the cuvette of a spectrophotometer, it was possible to monitor changes of pyruvate dehydrogenase activity continuously while the activities of the interconverting kinase and phosphatase could be independently manipulated. Permeabilized mitochondria were prepared from control and insulin-treated adipose tissue, and the properties of both the pyruvate dehydrogenase kinase and the phosphatase were compared in situ. No difference in kinase activity was detected, but increases in phosphatase activity were observed in permeabilized mitochondria from insulin-treated tissue. Further studies showed that the main effect of insulin treatment was a decrease in the apparent Ka of the phosphatase for Mg2+, in agreement with earlier studies with mitochondria made permeable to Mg2+ by using the ionophore A23187 [Thomas, Diggle & Denton (1986) Biochem. J. 238, 83-91]. No effects of spermine were detected, although spermine diminishes the Ka of purified phosphatase preparations for Mg2+. Since effects of insulin on pyruvate dehydrogenase phosphatase activity are not evident in mitochondrial extracts, it is concluded that insulin may act by altering some high-Mr component which interacts with the pyruvate dehydrogenase system within intact or permeabilized mitochondria, but not when the mitochondrial membranes are disrupted. PMID- 3026349 TI - Species-dependent isoenzyme subtypes of membrane-bound cyclic AMP-dependent protein kinase in highly purified cardiac sarcolemma. AB - Highly purified sarcolemma from dog and pig cardiac muscle has been shown to contain significant activities of a membrane-bound cyclic AMP-dependent protein kinase. In addition, these membranes undergo endogenous phosphorylation when incubated with Mg2+ and [gamma-32P]ATP. By comparing 32P-labelled patterns obtained with [gamma-32P]ATP and the photoaffinity label 8-azidoadenosine 3':5' [32P]monophosphate (8-azido-cyclic [32P]AMP), we have demonstrated that, whereas the major kinase isoenzyme in dog sarcolemma was Type II, that in the pig membrane was the Type I isoenzyme. PMID- 3026351 TI - The phosphoinositides exist in multiple metabolic pools in rabbit platelets. AB - The labelling of the phosphoinositides and phosphatidic acid in washed rabbit platelets incubated with [32P]phosphate or [3H]glycerol was studied in the presence of isotope and after unincorporated isotope had been removed. With both isotopes the increase in the specific radioactivity of phosphatidylinositol 4,5 bisphosphate (PIP2) lagged behind that of phosphatidylinositol 4-phosphate (PIP) but the specific radioactivity remained higher after unincorporated isotope had been removed. This result was consistent with the presence of a second pool of PIP2, which interconverted slowly with the pool of PIP2 which was in direct equilibrium with PIP, proposed to explain the increase in specific radioactivity of PIP2 which accompanies the decrease in amount of PIP2 at 10 s in ADP stimulated platelets. In platelets labelled with [3H]glycerol, the specific radioactivity of PIP2 became higher than that of PIP and the specific radioactivity of PIP became higher than that of phosphatidylinositol (PI). These results were interpreted to indicate that there were two pools of PIP; of these the pool with the higher specific radioactivity was the precursor of PIP2. Similarly, two pools of PI were proposed. The presence of pools of the phosphoinositides with different specific radioactivities necessitates the measurement of chemical amount of these compounds when studying the effect of stimulation of the platelets, since changes in labelling may not accurately reflect changes in the amount of the phosphoinositides. PMID- 3026350 TI - Sensitivity of adipocyte lipolysis to stimulatory and inhibitory agonists in hypothyroidism and starvation. AB - The responsiveness of lipolysis to the stimulatory agonists noradrenaline, corticotropin and glucagon and to the inhibitory agonists N6 phenylisopropyladenosine, prostaglandin E1 and nicotinic acid was investigated with rat white adipocytes incubated with a high concentration of adenosine deaminase (1 unit/ml). The cells were obtained from fed or 48 h-starved euthyroid animals or from fed or starved animals rendered hypothyroid by 4 weeks of treatment with low-iodine diet and propylthiouracil. Hypothyroidism increased sensitivity to and efficacy of all three inhibitory agonists in their opposition of noradrenaline-stimulated lipolysis. Starvation decreased sensitivity to all three inhibitory agonists when opposing basal lipolysis. Hypothyroidism decreased sensitivity to noradrenaline, glucagon and corticotropin by 37-, 4- and 4-fold respectively and decreased the maximum response to these agonists by approx. 50%, 50% and 75% respectively. Starvation reversed decreases in maximum response to these agonists in hypothyroidism. Starvation in the euthyroid state increased sensitivity to glucagon and noradrenaline, but did not alter sensitivity to corticotropin. Cells from hypothyroid rats were relatively insensitive to Bordetella pertussis toxin, which substantially increased basal lipolysis in the euthyroid state. PMID- 3026353 TI - The labelling of polyphosphoinositides with [32P]Pi and the accumulation of inositol phosphates in vasopressin-stimulated hepatocytes. AB - When hepatocytes were incubated with [32P]Pi, the kinetics for the labelling of the monoester phosphate groups of phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate were similar to each other and slightly slower than that for the labelling of the gamma-phosphate of ATP. Analysis of the water-soluble 3H-labelled materials derived from [3H]inositol-labelled hepatocytes revealed that, in addition to inositol and its mono-, bis- and tris phosphates (Ins, InsP, InsP2 and InsP3), these cells contained two unidentified radioactive compounds which co-eluted with InsP on anion-exchange chromatography. When [3H]inositol-labelled hepatocytes were stimulated with 0.23 microM vasopressin in the presence of 10 mM-Li+, there was an accumulation of radioactivity in InsP, InsP2 and InsP3 but not in Ins or the two unidentified compounds. Further analysis of these inositol phosphates by h.p.l.c. revealed that vasopressin also stimulates the accumulation of inositol tetrakisphosphate (InsP4) in these cells. Vasopressin-stimulated InsP and InsP2 accumulations were maximal in the presence of 1-10 mM-Li+ but InsP3 accumulation continued to increase up to 50 mM-Li+. Accumulated inositol phosphates were retained within the cell. Li+ from 1 to 50 mM did not influence the extent of vasopressin stimulated inositol lipid degradation in hepatocytes. In the absence of Li+, radioactivity in vasopressin-stimulated hepatocytes accumulated almost entirely in free inositol. The vasopressin-stimulated accumulation of inositol phosphates in the presence of 10 mM-Li+ was abolished by a V1-vasopressin antagonist. Inositol phosphate accumulation was not influenced by ionophore A23187, dimethyl sulphoxide or indomethacin. PMID- 3026355 TI - The radiosensitivity of rat thymocytes. AB - gamma-Irradiation in vitro apparently blocked a plasma-membrane associated, superoxide-producing, NADPH oxidase in rat thymocytes. Differential centrifugation of the mixed thymocytes indicated the smaller lymphocytes (approx. 6 microns diameter) to be the radiosensitive population. The oxidase system co isolated in part with thymus nuclei and could be solubilized by detergent treatment [Bellavite, Jones, Cross, Papini & Rossi (1984) Biochem. J. 223, 639 648]. Endogenous NADPH was the rate-limiting component for superoxide formation in vitro. The level of NADPH was lowered by gamma-irradiation, an effect mimicked by GSSG in the presence of 50 microM-ZnCl2 to inhibit GSSG reductase. These findings are suggested as the metabolic basis for interphase death of small lymphocytes exposed to ionizing radiation. PMID- 3026352 TI - The role of polyphosphoinositides and their breakdown products in A23187-induced release of arachidonic acid from rabbit polymorphonuclear leucocytes. AB - Stimulation of rabbit polymorphonuclear leucocytes with A23187 causes phospholipase C mediated breakdown of polyphosphoinositides, as evidenced by accumulation of [3H]inositol-labelled inositol bisphosphate and inositol trisphosphate. At the same time the polyphosphoinositides and the products of their breakdown, diacylglycerol and phosphatidic acid, label rapidly with radioactive arachidonic acid. Enhancement of polyphosphoinositide labelling is not as great as enhancement of diacylglycerol or phosphatidic acid labelling, suggesting additional early activation of a second independent synthetic pathway to the last named lipids. Experiments using double (3H/14C) labelling, to distinguish pools with different rates of turnover, suggest the major pool of arachidonic acid used for synthesis of lipoxygenase metabolites turns over more slowly than arachidonic acid in diacylglycerol, but at about the same rate as arachidonic acid esterified in phosphatidylcholine or phosphatidylinositol. Further, when cells are prelabelled with [14C]arachidonic acid, then stimulated for 5 min, it is only from phosphatidylcholine, and to a lesser extent phosphatidylinositol, that radiolabel is lost. Release of arachidonic acid is probably via phospholipase A2, since it is blocked by the phospholipase A2 inhibitor manoalide. The absence of accumulated lysophosphatides can be explained by reacylation and, in the case of lysophosphatidylinositol, deacylation. The importance of phospholipase A2 in phosphatidylinositol breakdown contrasts with the major role of phospholipase C in polyphosphoinositide hydrolysis. Measurements of absolute free fatty acid levels, as well as studies showing a correlation between production of radiolabelled hydroxyeicosatetraenoic acids and release of radiolabel from the phospholipid pool, both suggest that hydrolysis of arachidonic acid esterified into phospholipids is the limiting factor regulating formation of lipoxygenase metabolites. By contrast with A23187, fMet-Leu-Phe (a widely used polymorphonuclear leucocyte activator) is a poor stimulant for arachidonic acid release unless a 'second signal' (e.g. cytochalasin B, or a product of A23187-stimulated cells) is also present. In the presence of cytochalasin B, fMet-Leu-Phe, like A23187, stimulates release of radiolabelled arachidonic acid principally from phosphatidylcholine. PMID- 3026354 TI - Rapid formation of inositol 1,3,4,5-tetrakisphosphate and inositol 1,3,4 trisphosphate in rat parotid glands may both result indirectly from receptor stimulated release of inositol 1,4,5-trisphosphate from phosphatidylinositol 4,5 bisphosphate. AB - Addition of 1 mM-carbachol to [3H]inositol-labelled rat parotid slices stimulated rapid formation of [3H]inositol 1,3,4,5-tetrakisphosphate, the accumulation of which reached a peak 20 s after stimulation, and then declined rapidly towards a new steady state. The initial rate of formation of inositol 1,3,4,5 tetrakisphosphate was slower than that for inositol 1,4,5-trisphosphate. The radioactivity in [3H]inositol 1,3,4,5-tetrakisphosphate fell quickly in carbachol stimulated and then atropine-blocked parotid slices, suggesting that it is rapidly metabolized during stimulation. Parotid homogenates rapidly dephosphorylated inositol 1,4,5-trisphosphate, inositol 1,3,4,5-tetrakisphosphate and, less rapidly, inositol 1,3,4-trisphosphate. Inositol 1,3,4,5 tetrakisphosphate was specifically hydrolysed to a compound with the chromatographic properties of inositol 1,3,4-trisphosphate. The only 3H-labelled phospholipids that we could detect in parotid slices labelled with [3H]inositol for 90 min were phosphatidylinositol, phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate. Parotid homogenates synthesized inositol tetrakisphosphate from inositol 1,4,5-trisphosphate. This activity was dependent on the presence of ATP. We suggest that, during carbachol stimulation of parotid slices, the key event in inositol lipid metabolism is the activation of phosphatidylinositol 4,5-bisphosphate-specific phospholipase C. The inositol 1,4,5-trisphosphate thus liberated is metabolized in two distinct ways; by direct hydrolysis of the 5-phosphate to form inositol 1,4-bisphosphate and by phosphorylation to form inositol 1,3,4,5-tetrakisphosphate and hence, by hydrolysis of this tetrakisphosphate, to form inositol 1,3,4-trisphosphate. PMID- 3026356 TI - Purification and characterization of the phosphate/hydroxyl ion antiport protein from rat liver mitochondria. AB - The phosphate transport protein was purified from rat liver mitochondria by extraction in an 8% (v/v) Triton X-100 buffer followed by adsorption chromatography on hydroxyapatite and Celite. SDS/polyacrylamide-gel electrophoresis (10%, w/v) demonstrated that the purified polypeptide was apparently homogeneous when stained with Coomassie Blue and had a subunit Mr of 34,000. However, lectin overlay analysis of this gel with 125I-labelled concanavalin A demonstrated the presence of several low- and high-Mr glycoprotein contaminants. To overcome this problem, mitochondria were pre-extracted with a 0.5% (v/v) Triton X-100 buffer as an additional step in the purification of phosphate transport protein. SDS/polyacrylamide gradient gel electrophoresis (14 20%, w/v) of the hydroxyapatite and Celite eluates revealed one major band of Mr 34,000 when stained with Coomassie Blue. The known thiol group sensitivity of the phosphate transporter was employed to characterize the isolated polypeptide further. Labelling studies with N-[2-3H]ethylmaleimide showed that only the 34,000-Mr band was labelled in both the hydroxyapatite and Celite fractions, when purified from rat liver mitochondria. Further confirmation of its identity has been provided with an antiserum directed against the 34,000-Mr protein. Specific partial inhibition of phosphate uptake, as measured by iso-osmotic swelling in the presence of (NH4)2HPO4, was achieved when mitoplasts (mitochondria minus outer membrane) were incubated with this antiserum. Finally, amino acid analysis of the rat liver mitochondrial phosphate/hydroxyl ion antiport protein indicates that it is similar in composition to the equivalent protein isolated from ox heart. PMID- 3026359 TI - 4 beta-Phorbol 12-myristate 13-acetate attenuates the glucagon-induced increase in cytoplasmic free Ca2+ concentration in isolated rat hepatocytes. AB - Hepatocytes were isolated from rats and then loaded with the fluorescent Ca2+ indicator quin2. Glucagon caused a sustained increase (at least 5 min) in the fluorescence of the quin2-loaded cells; the increase was much greater than that observed with control, non-quin2-loaded, cells. These observations indicate that glucagon caused an increase in cytoplasmic free Ca2+ concentration [( Ca2+]c). The effects of glucagon were mimicked if forskolin (to activate adenylate cyclase), dibutyryl cyclic AMP or bromo cyclic AMP were added directly to the cells. Thus an increase in cyclic AMP concentration may mediate the effect of glucagon on [Ca2+]c. If 4 beta-phorbol 12-myristate 13-acetate (PMA; an activator of protein kinase C) was added to the cells before glucagon, the magnitude of the increase in [Ca2+]c was greatly diminished. If PMA was added after glucagon it caused a lowering of [Ca2+]c. These effects of PMA on the glucagon-induced increase in [Ca2+]c could not be mimicked if [Ca2+]c was increased by the Ca2+ ionophore ionomycin. Thus an event involved in the mechanism by which glucagon increases [Ca2+]c appears to be required for the action of PMA. If [Ca2+]c was increased by forskolin, dibutyryl cyclic AMP or bromo cyclic AMP, the effect of PMA on [Ca2+]c was similar to that observed when glucagon was used to elevate [Ca2+]c. When [Ca2+]c was raised by dibutyryl cyclic AMP the presence of the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine did not prevent the subsequent addition of PMA from causing [Ca2+]c to decrease. These observations suggest that PMA can inhibit the cyclic AMP-induced increase in [Ca2+]c independently of any changes in cyclic AMP concentration. Glucagon appears to increase [Ca2+]c by releasing intracellular stores of Ca2+ and stimulating net influx of Ca2+ into the cell; PMA greatly diminishes both of these effects. PMID- 3026357 TI - Luteinizing hormone stimulates the formation of inositol trisphosphate and cyclic AMP in rat granulosa cells. Evidence for phospholipase C generated second messengers in the action of luteinizing hormone. AB - The following studies were conducted to determine whether luteinizing hormone (LH), a hormone which increases cellular levels of cyclic AMP, also provokes increases in 'second messengers' derived from inositol lipid metabolism (i.e. inositol phosphates and diacylglycerol). Rat granulosa cells isolated from mature Graafian follicles were prelabelled for 3 h with myo-[2-3H]inositol. LH provoked rapid (5 min) and sustained (up to 60 min) increases in the levels of inositol mono-, bis, and trisphosphates (IP, IP2 and IP3, respectively). Time course studies revealed that IP3 was formed more rapidly than IP2 and IP following LH treatment. The response to LH was concentration-dependent with maximal increases at LH concentrations of 1 microgram/ml. LiCl (2-40 mM) enhanced the LH-provoked accumulation of all [3H]inositol phosphates, presumably by inhibiting the action of inositol phosphate phosphatases. The effectiveness of LH, however, was dependent on the concentration of lithium employed; maximal increases in IP were observed at 10 mM-LiCl, whereas maximal increases in IP2 and IP3 were observed at 20 mM- and 40 mM-LiCl, respectively. The stimulatory effects of LH on inositol phosphate and progesterone accumulation were also compared with changes in cyclic nucleotide levels. LH rapidly increased levels of inositol phosphates, progesterone and cyclic AMP, but transiently reduced levels of cyclic GMP. These results demonstrate that LH increases both cyclic AMP and inositol trisphosphate (and presumably diacylglycerol) in rat granulosa cells. Our findings suggest that two messenger systems exist to mediate the action of LH in granulosa cells. PMID- 3026358 TI - Synergistic stimulation of Ca2+ uptake by glucagon and Ca2+-mobilizing hormones in the perfused rat liver. A role for mitochondria in long-term Ca2+ homoeostasis. AB - A perfused liver system incorporating a Ca2+-sensitive electrode was used to study the long-term effects of glucagon and cyclic AMP on the mobilization of Ca2+ induced by phenylephrine, vasopressin and angiotensin. At 1.3 mM extracellular Ca2+ the co-administration of glucagon (10 nM) or cyclic AMP (0.2 mM) and a Ca2+-mobilizing hormone led to a synergistic potentiation of Ca2+ uptake by the liver, to a degree which was dependent on the order of hormone administration. A maximum net amount of Ca2+ influx, corresponding to approx. 3800 nmol/g of liver (the maximum rate of influx was 400 nmol/min per g of liver), was induced when cyclic AMP or glucagon was administered about 4 min before vasopressin and angiotensin. These changes are over an order of magnitude greater than those induced by Ca2+-mobilizing hormones alone [Altin & Bygrave (1985) Biochem. J. 232, 911-917]. For a maximal response the influx of Ca2+ was transient and was essentially complete after about 20 min. Removal of the hormones was followed by a gradual efflux of Ca2+ from the liver over a period of 30-50 min; thereafter, a similar response could be obtained by a second administration of hormones. Dose-response measurements indicate that the potentiation of Ca2+ influx by glucagon occurs even at low (physiological) concentrations of the hormone. By comparison with phenylephrine, the stimulation of Ca2+ influx by vasopressin and angiotensin is more sensitive to low concentrations of glucagon and cyclic AMP, and can be correlated with a 20-50 fold increase in the calcium content of mitochondria. The reversible uptake of such large quantities of Ca2+ implicates the mitochondria in long-term cellular Ca2+ regulation. PMID- 3026360 TI - Comparison of the processes involved in reduction by the substrate for two homologous flavocytochromes b2 from different species of yeast. AB - A detailed study of the electron exchanges involved between FMN and haem b2 groups within flavocytochrome b2 of yeast Hansenula anomala (H-enzyme) was performed. The results were compared with those for the homologous enzyme of yeast Saccharomyces cerevisiae (Sx-enzyme) re-investigated at 5 degrees C. The mid-point reduction potentials of FMN and haem were determined by two complementary methods: potentiometric titration with substrate, L-lactate, in the presence of dye mediators with quantification of the reduced species performed by spectrophotometry at suitable wavelengths; anaerobic titration of the enzyme by its substrate by monitoring the e.p.r. signals of the semiquinone and Fe3+ species. Values of Em,7 = -19, -23 and -45 V were determined respectively from the data for the three redox systems Ho/Hr, Fo/Fsq and Fsq/Fr in the H-enzyme instead of +6, -44 and -57 mV respectively in the Sx-enzyme [Capeillere-Blandin, Bray, Iwatsubo & Labeyrie (1975) Eur. J. Biochem. 54, 549-566]. Parallel e.p.r rapid-freezing and absorbance stopped-flow studies allowed determination of the time courses of the various redox species during their reduction by L-lactate. The flavin and the haem reduction time courses were biphasic. In the initial fast phase the reduction of flavin monitored by absorbance measurements is accomplished with a rate constant kF = 360 s-1. The reduction of the haem lags the reduction of flavin with a rate constant kH = 170 s-1. The appearance of flavin free radical is slower than the reduction in flavin absorbance and occurs with a rate constant close to that of the reduction of the haem. At saturating L lactate concentration the initial rapid phase (up to 15 ms) involved in the overall turnover can be adequately simulated with a two-step reaction scheme. The main difference between the enzymes lies especially at the level of the first step of electron exchange between bound lactate and flavin, which for the H enzyme is no longer the rate-limiting step in the haem reduction and becomes 8 fold faster than in the Sx-enzyme. Consequently in the H-enzyme for the following step, the intramolecular transfer from flavin hydroquinone to oxidized haem, a reliable evaluation of the rate constants becomes possible. Preliminary values are k+2 = 380 s-1 and k-2 = 120 s-1 at 5 degrees C.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3026362 TI - Abnormal regulation of insulin secretion in the genetically obese (ob/ob) mouse. AB - To determine whether the abnormal insulin-secretory activity encountered in obese mice is due to an anomaly in the production of cyclic AMP, islets of lean and obese mice were incubated with forskolin under various conditions. Our data show that, in addition to the well-known quantitative differences in insulin-secretory activity between islets of lean and obese mice, there are important qualitative differences. The islets of obese mice accumulated less cyclic AMP than did those of lean mice in response to given doses of forskolin, yet their insulin secretion was enhanced to much higher values. In the islets of obese mice, but not in those of lean mice, the stimulatory effect of forskolin on insulin secretion was evident even at non-stimulatory concentrations of glucose or in Ca2+-deprived incubation media, showing that the islet of the obese mouse is less dependent on a primary stimulus (glucose) and on the provision of normal concentrations of Ca2+ in the bathing medium than is the islet of the lean mouse. PMID- 3026361 TI - Effects of Ca2+ on phosphoinositide breakdown in exocrine pancreas. AB - Recent studies have established that inositol 1,4,5-trisphosphate [I(1,4,5)P3] provides the link between receptor-regulated polyphosphoinositide hydrolysis and mobilization of intracellular Ca2+. Here, we report the effects of Ca2+ on inositol trisphosphate (IP3) formation from phosphatidylinositol bisphosphate (PIP2) catalysed by phospholipase C in intact and electrically permeabilized rat pancreatic acinar cells. In permeabilized cells, the Ca2+-mobilizing agonist caerulein stimulated [3H]IP3 formation when the free [Ca2+] was buffered at 140 nM, the cytosolic free [Ca2+] of unstimulated pancreatic acinar cells. When the free [Ca2+] was reduced to less than 10 nM, caerulein did not stimulate [3H]IP3 formation. Ca2+ in the physiological range stimulated [3H]IP3 formation and reduced the amount of [3H]PIP2 in permeabilized cells. The effects of Ca2+ and the receptor agonist caerulein were additive, but we have not established whether this reflects independent effects on the same or different enzymes. The effect of Ca2+ on [3H]IP3 formation by permeabilized cells was unaffected by inhibitors of the cyclo-oxygenase and lipoxygenase pathways of arachidonic acid metabolism; nor were the effects of Ca2+ mimicked by addition of arachidonic acid. These results suggest that the effects of Ca2+ on phospholipase C activity are not a secondary consequence of Ca2+ activation of phospholipase A2. Changes in free [Ca2+] (less than 10 nM-1.2 mM) did not affect the metabolism of exogenous [3H]I(1,4,5)P3 by permeabilized cells. In permeabilized cells, breakdown of exogenous [3H]IP3 to [3H]IP2 (inositol bisphosphate), and formation of [3H]IP3 in response to receptor agonists were equally inhibited by 2,3-bisphosphoglyceric acid. This suggests that the [3H]IP2 formed in response to receptor agonists is entirely derived from [3H]IP3. In intact cells, [3H]IP3 formation was stimulated when ionomycin was used to increase the cytosolic free [Ca2+]. However, a maximal concentration of caerulein elicited ten times as much IP3 formation as did the highest physiologically relevant [Ca2+]. We conclude that the major effect of receptor agonists on IP3 formation does not require an elevation of cytosolic free [Ca2+], although the increase in free [Ca2+] that normally follows IP3 formation may itself have a small stimulatory effect on phospholipase C. PMID- 3026363 TI - The ability of salts to inhibit the reaction between periodate anions and ovotransferrin. AB - The reaction between periodate anions and apo-ovotransferrin results in the rapid abolition of the iron-binding ability of the protein and the loss of approximately 4 mol of tyrosine/mol of protein. The degree of inhibition exerted by a variety of salts on the rate of this reaction is found to be inconsistent with the lyotropic series and suggests the existence of a complex anion-binding site in the apoprotein. The existence of this site may explain the action of periodate anions on ovotransferrin. PMID- 3026364 TI - Identification of the phosphatase deinhibitor protein phosphatases in rabbit skeletal muscle. AB - In rabbit skeletal muscle the polycation-stimulated (PCS) protein phosphatases [Merlevede (1985) Adv. Protein Phosphatases 1, 1-18] are the only phosphatases displaying significant activity toward the deinhibitor protein. Among them, the PCSH protein phosphatase represents more than 80% of the measurable deinhibitor phosphatase activity associated with the PCS phosphatases. The deinhibitor phosphatase activity co-purifies with the PCSH phosphatase to apparent homogeneity. In the last purification step two forms of PCSH phosphatase were separated (PCSH1, containing 62, 55 and 34 kDa subunits, and PCSH2, containing 62 and 35 kDa subunits), both showing the same deinhibitor/phosphorylase phosphatase activity ratio. The activity of the PCSH phosphatase toward the deinhibitor is not stimulated by polycations such as protamine, histone H1 or polylysine, unlike the stimulation observed with phosphorylase as the substrate. The phosphorylase phosphatase activity of PCSH phosphatase is inhibited by ATP, PPi and Pi, whereas the deinhibitor phosphatase activity of the enzyme is much less sensitive to these agents. PMID- 3026365 TI - The structure of the hepatic insulin receptor and insulin binding. AB - Hepatocytes or hepatic plasma membranes were photoaffinity-labelled with radioiodinated N epsilon B29-monoazidobenzoyl-insulin. Analysis of the samples by SDS/polyacrylamide-gel electrophoresis and autoradiography revealed the insulin receptor as a predominant band of 450 kDa. When hepatic plasma membranes were first treated with clostridial collagenase and then photolabelled, the insulin receptor appeared as a predominant band of 360 kDa. This effect of collagenase treatment on the insulin receptor was due to Ca2+-dependent heat-labile proteinases contaminating the preparation of collagenase, and it could be mimicked by elastase. The decrease in size of the insulin receptor to 360 kDa resulted from the loss of a receptor component that was inaccessible to photolabelling. In contrast, the size of the insulin receptor of intact cells was not affected by collagenase treatment. This suggests that the site sensitive to proteolysis was located on the cytoplasmic side of the plasma membrane. In hepatic plasma membranes that were treated with collagenase or elastase, and contained the 360 kDa form of the insulin receptor, the binding affinity for insulin was increased by up to 2-fold. These findings support the concept that a component which is either a part of, or closely associated with, the insulin receptor may regulate its affinity for insulin. PMID- 3026366 TI - [Phosphotyrosine]protein phosphatase in rat brain. A major [phosphotyrosine]protein phosphatase is a 23 kDa protein distinct from acid phosphatase. AB - A [phosphotyrosine]protein phosphatase (PTPPase) was purified almost to homogeneity from rat brain, with [32P]p130gag-fps, an oncogene product of Fujinami sarcoma virus, as substrate. The characteristics of the purified preparation of PTPPase were as follows: the enzyme was a monomer with a molecular mass of 23 kDa; its optimum pH was 5.0-5.5; its activity was not dependent on bivalent cations; its activity was strongly inhibited by sodium vanadate, but was not inhibited by ZnCl2, L(+)-tartrate or NaF; it catalysed the dephosphorylation of [32P]p130gag-fps, [[32P]Tyr]casein, p-nitrophenyl phosphate and L phosphotyrosine, but did not hydrolyse [[32P]Ser]tubulin, L-phosphoserine, DL phosphothreonine, 5'-AMP, 2'-AMP or beta-glycerophosphate significantly. During the purification, most of the PTPPase activity was recovered in distinct fractions from those of conventional low-molecular-mass acid phosphatase (APase), which was reported to be a major PTPPase [Chernoff & Li (1985) Arch. Biochem. Biophys. 240, 135-145], from DE-52 DEAE-cellulose column chromatography, and those two enzymes could be completely separated by Sephadex G-75 column chromatography. APase also showed PTPPase activity with [32P]p130gag-fps, but the specific activity was lower than that of PTPPase with molecular mass of 23 kDa, and it was not sensitive to sodium vanadate. These findings suggested that PTPPase (23 kDa) was the major and specific PTPPase in the cell. PMID- 3026367 TI - Superoxide-dependent and -independent mechanisms of iron mobilization from ferritin by xanthine oxidase. Implications for oxygen-free-radical-induced tissue destruction during ischaemia and inflammation. AB - Xanthine oxidase is able to mobilize iron from ferritin. This mobilization can be blocked by 70% by superoxide dismutase, indicating that part of its action is mediated by superoxide (O2-). Uric acid induced the release of ferritin iron at concentrations normally found in serum. The O2(-)-independent mobilization of ferritin iron by xanthine oxidase cannot be attributed to uric acid, because uricase did not influence the O2(-)-independent part and acetaldehyde, a substrate for xanthine oxidase, also revealed an O2(-)-independent part, although no uric acid was produced. Presumably the amount of uric acid produced by xanthine oxidase and xanthine is insufficient to release a measurable amount of iron from ferritin. The liberation of iron from ferritin by xanthine oxidase has important consequences in ischaemia and inflammation. In these circumstances xanthine oxidase, formed from xanthine dehydrogenase, will stimulate the formation of a non-protein-bound iron pool, and the O2(-)-produced by xanthine oxidase, or granulocytes, will be converted by 'free' iron into much more highly toxic oxygen species such as hydroxyl radicals (OH.), exacerbating the tissue damage. PMID- 3026368 TI - The presence and activity in normal and regenerating rat liver postmicrosomal supernatant fraction of an enzyme with properties similar to those of membrane bound 5'-nucleotidase. AB - An alkaline 5'-nucleotidase with properties similar to those of membrane-bound 5' nucleotidase was recovered in soluble form in the postmicrosomal supernatant fraction (cytosol) of rat liver. The enzyme seems to constitute a quantitatively distinct fraction, since the activity in postmicrosomal supernatants was increased by a further 10% by additional homogenization of livers. Lysosomal acid phosphatase activity increased similarly, whereas other membrane-bound marker enzymes alkaline phosphatase, phosphodiesterase I and glucose-6-phosphatase showed no increase when homogenization of liver tissue was continued. Gel permeation chromatography and pH-dependence studies indicated that enzyme activity in the supernatant fraction with 0.3 mM-UMP or -AMP as substrate at pH 8.1 was about 85 or 100% specific respectively. In regenerating liver the enzyme recovered in soluble form showed decreased specific activity, in contrast with alkaline phosphatase measured for comparison. The nucleotidase activity per mg of cytosolic protein was 2.1 nmol/min with AMP as substrate. The total activity measured in the postmicrosomal supernatant was 1.5% of the homogenate activity measured in the presence of detergent. PMID- 3026369 TI - The phosphorylation of ribosomal protein S6 by protein kinases from cells infected with pseudorabies virus. AB - We examined the ability of protein kinase activities from BHK (baby-hamster kidney) cells infected with pseudorabies virus to catalyse the phosphorylation of ribosomal protein S6 in vitro. When the cytosol from infected cells was fractionated on DEAE-cellulose, 40S ribosomal protein kinase activity was found associated with the two isoforms of the cyclic AMP-dependent protein kinase, protein kinase C and a protein kinase (ViPK, virus-induced protein kinase) only detected in infected cells. The phosphorylation of ribosomal protein by ViPK was of particular interest because the appearance of the protein kinase and the increase in the phosphorylation of protein S6 in infected cells shared a similar time course. At moderate concentrations of KCl the major ribosomal substrate for ViPK was ribosomal protein S7, a protein not found to be phosphorylated in vivo. However, at 600 mM-KCl, or in the presence of 5-10 mM-spermine at 60-150 mM-KCl, the phosphorylation of ribosomal protein S7 was suppressed and ribosomal protein S6 became the major substrate. The maximum stoichiometry of phosphorylation obtained under the latter conditions was 1-2 mol of phosphate/mol of S6, and only mono- and di-phosphorylated forms of S6 were detected on two-dimensional gel electrophoresis. As the infection of BHK cells by pseudorabies virus results in the appearance of phosphorylated species of S6 containing up to 5 mol of phosphate/mol of S6 protein, it appears unlikely that ViPK alone can be responsible for the multiple phosphorylation seen in vivo. Nevertheless, tryptic phosphopeptide analysis did indicate that in vitro ViPK catalysed the phosphorylation of at least one of the sites on ribosomal protein S6 phosphorylated in vivo, so that a contributory role for the enzyme in the phosphorylation in vivo cannot be excluded. PMID- 3026371 TI - Purification and characterization of the active fragment from Bacillus thuringiensis delta-toxin. AB - Limited tryptic hydrolysis of a partially purified delta-toxin (Mr = 100,000) from Bacillus thuringiensis, has produced a polypeptide fragment of Mr = 60,000 containing the full biological activity. The fragment was the only polypeptide observed in the polyacrylamide-gel electrophoresis of the delta-toxin after treatment with trypsin and could be purified by DEAE-cellulose chromatography. Amino acid and partial sequence analyses indicate that the 60,000 Mr fragment has been derived from the mid-section of the holotoxin peptide; over 80% of Lys, 65% of Pro and 50% of His residues in the holotoxin have been lost in the active fragment. This section must contain the active site since its specific insecticidal activity is approximately twice that of the holotoxin. The active fragment shows complete cross-reactivity with the antiserum raised against the native toxin, and appeared to possess higher thermal stability than the mother protein. It provides a powerful tool for studies of the structure involved in the insecticidal activity. PMID- 3026370 TI - The relationship between intra- and extra-cellular surfactant phospholipids in the lungs of rabbits and the effects of silica-induced lung injury. AB - Extensive homogenization of lung tissue by nitrogen decompression in a Parr disruption bomb increased by 5-fold the yields of low-density phospholipid (d = 1.06) achieved by other methods. This intracellular phospholipid preparation was high in phosphatidylcholines (84.3%), particularly disaturated phosphatidylcholine (51.2%). On the basis of its low density, composition, and morphological appearance, we concluded that this phospholipid was derived from the intracellular compartment of pulmonary surfactant. We examined the relationship between intra- and extra-cellular surfactant pools according to age, gender and silica-induced pulmonary injury. In normal animals the intracellular pool of surfactant phospholipids increased from 1.54 +/- 0.14 mg at 1 day after birth to 62.30 +/- 4.50 mg per pair of lungs after 31 months, and over the same time period the extracellular pool increased from 1.04 +/- 0.15 mg to 27.45 +/- 2.30 mg per pair of lungs. The ratio between the extracellular and intracellular pools of surfactant increased from 1.50 +/- 0.19 at 1 day after birth to 2.28 +/- 0.23 after 31 months of age. The ratio between the two pools was not influenced by gender, but was changed by the intratracheal injection of silica into the lungs. Intratracheal injection of silica dust increased the levels of surfactant in both compartments, but not to the same extent, indicating that the ratio between the pools could be changed by toxic materials. These data suggest the existence of a size relationship between the intra- and the extra-cellular pools of surfactant, a relationship which implies a common regulatory mechanism that can be disturbed during pulmonary injury. PMID- 3026372 TI - A phorbol ester and 1-oleoyl-2-acetylglycerol induce Na+/H+ exchange in human platelets. AB - This study aimed at investigating the mechanisms by which stimulation of human platelets results in activation of Na+/H+ exchange. Platelets were suspended in a slightly buffered medium and the stimulus-induced, amiloride-sensitive H+ release, reflecting Na+/H+ exchange, was estimated from changes in the medium pH. H+ release could be evoked by thrombin and by activators of protein kinase C such as 1-oleoyl-2-acetylglycerol (OAG) or 12-O-tetradecanoylphorbol-13-acetate (TPA). Both the thrombin-and the OAG-induced Na+/H+ exchange could be blocked by trifluoperazine, a protein kinase C inhibitor. The thrombin-induced H+ release was also sensitive to increased intracellular cAMP levels, probably due to inhibition of phospholipase C activation, whereas the OAG-induced activation of Na+/H+ exchange was unaffected. Our data suggest that activation of Na+/H+ exchange is mediated by protein kinase C. PMID- 3026373 TI - Suppression of triglyceride secretion by epinephrine in isolated rat hepatocytes. AB - The effect of epinephrine on triglyceride synthesis and secretion was examined in isolated rat hepatocytes. Epinephrine potently inhibited triglyceride secretion but did not affect cellular triglyceride content or the rate of incorporation of radiolabelled glycerol into cell triglyceride. The inhibitory effect of epinephrine was abolished by inclusion of the alpha-adrenergic antagonist prazosin but not the beta-antagonist propranolol. PMID- 3026374 TI - Evidence that neomycin inhibits HSV 1 infection of BHK cells. AB - The effect of neomycin on the Herpes Simplex Virus (HSV) type 1 and 2 infection of baby hamster kidney cells was studied. Neomycin concentrations of 3 mM or more caused a more than 90% inhibition of HSV 1 proliferation, while it had no effect on HSV 2 proliferation, measured as plaque-forming units. Furthermore, neomycin must be present at the time of infection in order to exert full effect, addition 1 hour postinfection was comparable to untreated cells. This indicates that neomycin may specifically interfere with very early stages of HSV 1 infection. PMID- 3026376 TI - Purification of vanadium binding substance from the blood cells of the tunicate, Ascidia sydneiensis samea. AB - Vanadium binding substance has been partially purified through chromatographies on Sephadex G-25 and SE-Cellulose at pH 2.3. The binding substance was colorless, relatively stable and maintained vanadium ion. The vanadium ion in the substance existed in vanadyl form (VO(IV)). Furthermore, the substance had an apparent affinity for exogenous vanadium ion(V) and contained a reducing sugar. PMID- 3026375 TI - Pyrimidine dimer dependent cleavage of single-stranded DNA by T4 UV endonuclease. AB - T4 UV endonuclease cleaves double- and single-stranded DNA with equal specificity for photo-pyrimidine dimers. Thus, the enzyme can be used for mapping and quantifying pyrimidine dimers in single-stranded DNA as well as in double stranded DNA. Mapping of pyrimidine dimers shows that rates of UV-dimerization are not only affected by 5', 3' adjacent bases, but also by position within pyrimidine tracts. Di-pyrimidines at 3' ends of tracts are more photoreactive than those at 5' ends. PMID- 3026377 TI - Evidence for the presence of functional beta-adrenoceptor along the proximal tubule of the rat kidney. AB - Evidence for the presence of beta adrenoceptors on proximal tubules from the rat kidney has been obtained using enriched tubule suspensions prepared by Percoll centrifugation. Intact tubules demonstrated simultaneous enrichment of parathyroid hormone and isoproterenol sensitive cAMP production with no enrichment of antidiuretic hormone sensitive cAMP production. Both norepinephrine and epinephrine were less potent than isoproterenol and the stimulatory effect of catecholamines could be blocked with propranolol but not phentolamine. The stimulatory effect of norepinephrine on cellular phenylalanine uptake is blunted by co-addition of isoproterenol suggesting that the beta receptor may modulatory catecholamine stimulated transport. PMID- 3026379 TI - An ESR study on lipid peroxidation process. Formation of hydrogen atoms and hydroxyl radicals. AB - Lipid peroxidation process was studied by the combination of ESR spectroscopy and spin trapping with 5,5-dimethyl-1-pyrroline-N-oxide. Autoxidation of egg lecithin phosphatidylcholine dispersed in water was found to produce hydrogen atoms and hydroxyl radicals. It was also found that for producing such reactive species, unsaturated fatty acid moieties are needed. The ESR spectrum obtained from this model system was identical with that from body fluid such as serum, indicating that hydrogen atoms and hydroxyl radicals could be generated in living bodies during the course of lipid peroxidation. PMID- 3026378 TI - GTP regulation of platelet-activating factor binding to human neutrophil membranes. AB - Radiolabeled ligand binding studies showed that specific receptors for platelet activating factor are present in human neutrophil membranes. GTP at 10(-7) to 10( 3) M decreased the specific binding of platelet-activating factor to neutrophil membranes in a dose-dependent manner. Inhibition of platelet-activating factor binding was also induced by other guanine nucleotides but not by adenine nucleotides. Our results suggest that platelet-activating factor receptor in human neutrophil membranes may be coupled to a guanine nucleotide binding protein. PMID- 3026380 TI - Monoclonal antibody that blocks phosphoinositide-dependent activation of mouse tumor DNA polymerase alpha. AB - A monoclonal antibody (MaB) against mouse sarcoma DNA polymerase alpha was isolated from the culture medium of an IgG-secreting hybridoma. The MaB demonstrated reactivity against two murine DNA polymerase alpha preparations and a calf thymus DNA polymerase alpha. Murine sarcoma polymerase was activated in vitro by phosphatidylinositol-4-monophosphate (PIP) showing increased deoxynucleotidyltransferase activity and enhanced binding affinity to activated DNA template. The MaB did not neutralize polymerase activity, but blocked further activation of the enzyme by PIP. Treatment of polymerase with MaB prior to treatment with PIP inhibited both increased enzyme activation and increased binding of the enzyme to DNA template. Treatment of polymerase with MaB subsequent to treatment with PIP did not block enzyme activation or increased DNA template binding. The data suggest that this anti-DNA polymerase alpha IgG is directed against a regulatory subunit of the polymerase rather than the catalytic subunit. The antibody may serve to distinguish between DNA polymerase alpha preparations with distinctly different regulatory subunits. PMID- 3026381 TI - Phosphorylation of the mouse hepatitis virus nucleocapsid protein. AB - Analysis of the radiolabeled tryptic peptides derived from the nucleocapsid proteins of two serotypes of mouse hepatitis virus showed each to have a small number of unique peptides; however, two biologically distinct variants of the JHM strain appeared identical. Analysis of [32P]-labeled nucleocapsid-derived peptides showed that phosphorylation occurs at only a few sites and that all three viruses differed in the sites of phosphorylation. No differences in the sites of phosphorylation were found between the nucleocapsid proteins derived from purified virions and the membranes or the cytosol of infected cells, suggesting that post-translational phosphorylation plays no role in the regulation of viral assembly. These data show unequivocal evidence that the nucleocapsid proteins of mouse hepatitis virus strains differ in the sites of phosphorylation. PMID- 3026382 TI - Modulation by phorbol myristate acetate of arachidonic acid release and leukotriene synthesis by human polymorphonuclear leukocytes stimulated with A23187. AB - Phorbol myristate acetate augmented the release of 3H-AA and the synthesis of leukotriene B4 and 5-hydroxyeicosatetraenoic acid by human polymorphonuclear leukocytes stimulated by A23187. PMA alone had no effect. Enhancement of the response to A23187 was not seen when the inactive phorbol ester 4-alpha phorbol didecanoate was added with A23187. These data are consistent with the hypothesis that activation of protein kinase C enhances AA release and metabolism in stimulated polymorphonuclear leukocytes. PMID- 3026383 TI - Use of double spin-labeled histones to monitor histone-chromatin integration. AB - Tyrosine-specific nitroxide spin labels have been synthesized that utilize either deuterium or deuterium and [15N] isotopic substitution within the nitroxide ring. These probes have been used to differentially spin label and simultaneously monitor both histones H1 and H5, during the displacement of endogenous spin labeled H1 from reconstituted chromatin by exogenously added spin-labeled H5. PMID- 3026384 TI - Pyrophosphate-dependent sucrose metabolism and its activation by fructose 2,6 bisphosphate in sucrose importing plant tissues. AB - In the presence of pyrophosphate and uridine diphosphate, sucrose was cleaved to form glucose 1-phosphate and fructose with soluble extracts from sucrose importing plant tissues. The glucose 1-phosphate then was converted through glycolysis to triose phosphates in a pyrophosphate-dependent pathway which was activated by fructose 2,6-bisphosphate. Much less activity, less than 5%, was found in sucrose exporting tissue extracts from the same plants. These findings suggest that imported sucrose is metabolized in the cytoplasm of plant tissues by utilizing pyrophosphate and that sucrose metabolism is partially regulated by fructose 2,6-bisphosphate. PMID- 3026386 TI - [17O]oxygen hyperfine structure for the hydroxyl and superoxide radical adducts of the spin traps DMPO, PBN and 4-POBN. AB - [17O]oxygen hyperfine coupling constants are reported for the superoxide and hydroxyl radical adducts with the spin traps 5,5-dimethyl-1-pyrroline N-oxide, N t-butyl-alpha-phenylnitrone and alpha-(4-pyridyl 1-oxide)-N-t-butylnitrone. These couplings provide spectroscopic evidence that the spin adducts have been correctly identified. PMID- 3026385 TI - A search for oxygen-centered free radicals in the lipoxygenase/linoleic acid system. AB - Studies of the oxygenation of linoleic acid by soybean lipoxygenase utilizing electron spin resonance spectroscopy and oxygen uptake have been undertaken. The spin trap, alpha-(4-pyridyl-1-oxide)-N-t-butylnitrone (4-POBN) was included in the lipoxygenase system to capture short-lived free radicals. Correlation of radical adduct formation rates with oxygen uptake studies indicated that the major portion of radical adduct formation occurred when the system was nearly anaerobic. Incubations containing [17O]oxygen with nuclear spin of 5/2 did not have additional ESR lines as would be expected if an oxygen-centered 4-POBN-lipid peroxyl radical adduct were formed indicating that the trapped radical must be reassigned as a carbon-centered species. To establish the presence of [17O2]oxygen in our incubations, a portion of the gas from the lipoxygenase/linoleate experiments was used to prepare the 4-POBN-superoxide radical adduct utilizing a superoxide producing microsomal/paraquat/NADPH system. PMID- 3026387 TI - Thrombin receptor occupancy initiates cell proliferation in the presence of phorbol myristic acetate. AB - A combination of DIP-thrombin and either PMA (50 ng/ml) or dioctanoyl glycerol stimulates DNA synthesis in serum free cultures of NIL hamster cells similar to that previously reported for the combinatory effect of DIP-thrombin and gamma thrombin. Thus, PMA or dioctanoyl glycerol appears to generate signals normally stimulated by gamma-thrombin interaction with cells. This stimulation was not observed when cells were treated with DIP-thrombin and 4-beta-phorbol or 4-alpha phorbol 12,13-didecanoate. Therefore, it appears that this effect is mediated through activation of protein kinase C and that this activation plays an important role in thrombin mitogenesis. PMID- 3026388 TI - Stimulation of phosphatidylinositol turnover by concanavalin A is not sufficient to activate mouse thymocytes. AB - Mouse thymocytes treated with the lectin Concanavalin A do not proliferate nor do they develop responsiveness to interleukin 2. Co-treatment with Concanavalin A and either lectin-activated splenocyte conditioned medium or phorbol ester caused increased interleukin 2 receptor expression and proliferation. Under these conditions, lectin alone stimulated a 3.4 fold increase in phosphatidylinositol turnover which was unaffected by the presence of conditioned medium. Phosphorylation of a 55 kD protein was stimulated in response to conditioned medium or phorbol ester, but not lectin. These results indicate that stimulation of phosphoinositide turnover is not sufficient to activate thymocytes, and suggest that costimulating factors activate a kinase which is distinct from protein kinase C, or alternatively, activate protein kinase C through a process which is not coupled to phosphoinositide turnover. PMID- 3026389 TI - Reversible shape change in platelets is regulated by phosphatidylinositol metabolism. AB - Reversible platelet activation was studied after mechanical activation by low speed centrifugation (600 xg). Immediately following centrifugation platelets exhibited no shape change response to low doses of thrombin, collagen and ADP. After incubation at 37 degrees C a time-dependent recovery of the shape change response was observed. This was accompanied by a 50% decrease of 32P incorporation into phosphatidic acid (PA) phosphatidylinositol-monophosphate (PIP), but not PIP2, relative to levels observed immediately after centrifugation. After 60 minutes platelet relaxation was complete: [32]PA and [32]PIP reached lowest levels and the shape change response to low doses of thrombin was completely restored. PMID- 3026390 TI - Degrading activity for human parathyroid hormone [PTH-(1-84)] in rat osteoblast like osteosarcoma cell line UMR106. AB - The degrading activity for human parathyroid hormone [hPTH-(1-84)] was studied in a rat osteoblast-like osteosarcoma cell line UMR106. At 37 C,UMR106 cells degraded hPTH-(1-84) into fragments in a time-dependent manner, which was shown by a radioimmunoassay with the use of antibody recognizing the C-terminal and middle regions of PTH molecule, whereas the degradation was completely suppressed at 4 C and failed to occur in the absence of the cells. The Lineweaver-Burk plot of this degrading activity at 37 C showed a fairly good linearity and gave a Km value of 5.1 X 10(-7) M. Reverse-phase high-performance liquid chromatography (HPLC) analysis of immunoreactive PTH fragments in the medium disclosed two peaks aside from intact PTH, indicating a limited PTH-hydrolyzing activity of UMR106 cells cleaving the molecule between at least two separate positions. This study suggests the possible involvement of osteoblasts on the metabolism of intact PTH. PMID- 3026391 TI - Molecular cloning of the structural gene for Acinetobacter citrate synthase. AB - The structural gene for citrate synthase of Acinetobacter anitratum has been cloned in Escherichia coli in a form which expresses the enzyme. A library of EcoRI fragments of Acinetobacter genomic DNA was prepared in the vector lambda gt10, and clones were screened by hybridization with an E. coli citrate synthase clone under conditions of reduced stringency. A 6.5 kbp clone was obtained which was subcloned into pBR322, and shown to direct the formation of Acinetobacter citrate synthase in E. coli hosts. The promoter was located within a BglII fragment, and from this information the orientation of the gene was deduced. PMID- 3026392 TI - Expression of human interleukin-2 receptor cDNA in E. coli. AB - cDNAs for human interleukin-2 receptor were recently cloned and sequenced (Leonard et al., 1984, Nature 311, 626-631; Nikaido et al., 1984, Nature 311, 631 635; Cosman et al., Nature 312, 768-771). In the studies reported here, we describe the expression of a cDNA clone for the human interleukin-2 receptor in E. coli using an "open reading frame" expression vector pMR100. The inserted cDNA was expressed in E. coli transformants as a tripartite fusion polypeptide fused to the lambda cI protein at its amino terminus and to beta-galactosidase at its carboxy terminus. We demonstrate that the bacterially produced IL-2 receptor protein can bind to IL-2. PMID- 3026393 TI - Effect of insulin on glucagon binding to isolated rat epididymal adipocytes. AB - Effect of insulin on glucagon binding to rat epididymal adipocytes was studied in vitro. [125I]iodoglucagon binding to isolated adipocytes was increased by preincubation of the cells with insulin. Maximal increase was observed with 7 X 10(-10) M insulin. In Scatchard analysis, [125I]iodoglucagon competition data generated one binding site with a single affinity for glucagon binding in the cells pretreated with buffer alone. Pretreatment of the cells with insulin increased the affinity without changes in the number of binding sites. [125I]iodoglucagon binding to isolated adipocytes was not affected by pretreatment of the cells with luteinizing hormone, follicle-stimulating hormone, growth hormone, or with prolactin. These results suggest that insulin stimulates glucagon binding to adipocytes. PMID- 3026394 TI - Rapid modulation of tumor necrosis factor membrane receptors by activators of protein kinase C. AB - Tumor necrosis factor membrane receptors are rapidly down-regulated upon treatment of activated T lymphocytes with various activators of protein kinase C. Loss of binding-capacity was half maximal after 2 min. incubation in 10 ng/ml of phorbol 12-myristate 13-acetate. A similar modulation could be induced with either the calcium ionophore A 23187 or the protein kinase C activator 1-oleyl-2 acetyl glycerol, whereas 1,2-diolein and dibutyryl cAMP were ineffective. Protein kinase C inhibitor H7 antagonizes the phorbol ester-induced TNF receptor modulation. These data suggest an important role of protein kinase C in the control of TNF responsiveness by regulation of TNF binding-capacity possibly via direct phosphorylation of specific receptor proteins. PMID- 3026395 TI - Possible involvement of germ cells in the regulation of oestradiol-17 beta and ABP secretion by immature rat Sertoli cells (in vitro studies). AB - The effects of germ cells prepared from adult rats and of media conditioned by some of these germ cells have been studied in vitro on both ABP and oestradiol-17 beta secretion by immature rat Sertoli cells. Addition of the germ cells to the Sertoli cell cultures resulted in both a dose-dependent increase of ABP secretion and a dose-dependent inhibition of oestradiol production. These effects were suppressed after removal of germ cells by hypotonic treatment. Furthermore, spent media of highly viable germ cells (SMGC), but not spent media of an epithelial cell line, mimicked the effects of germ cells themselves on ABP and oestradiol levels after FSH or dbcAMP stimulation. These effects were reversible when SMGC were replaced by fresh media and did not result from a change in the conversion of oestradiol to oestrone. SMGC effects were unaltered by heating at 60 degrees C for 30 min, by freezing and thawing and non dialysable (MW greater than 10,000). However, heating at 100 degrees C for 3 min and treatment by trypsin, suppressed the SMGC effects. This indicates that the stimulation of ABP and inhibition of oestradiol levels by germ cells, in vitro, could be mediated by factor(s) of proteinaceous nature. PMID- 3026396 TI - Chlorate--a potent inhibitor of protein sulfation in intact cells. AB - Chlorate is known to be an in vitro inhibitor of ATP-sulfurylase, the first enzyme in the biosynthesis of PAPS which is the ubiquitous co-substrate for sulfation. Here, the effect of chlorate on protein sulfation in intact cells was investigated. Treatment of various cell cultures with 1 mM sodium chlorate in a medium low in sulfate and sulfur-containing amino acids resulted in an inhibition of protein sulfation greater than 95%. Tyrosine as well as carbohydrate sulfation was blocked. Chlorate did not inhibit protein synthesis and did not exhibit any other toxic effects, even after prolonged treatment of cell cultures. Thus, chlorate treatment provides a powerful tool for studying the biological significance of protein sulfation. PMID- 3026397 TI - Activation of collagenase production in a rat macrophage-like cell line. AB - Collagenase activity from a rat macrophage line, AK-5, has been studied. The enzyme is present in these cells and its release is stimulated by Con A, LPS and a lymphokine preparation from rat spleen cells. Collagenase activity from these cells is inhibited by EDTA, cysteine or dithiothreitol, thereby suggesting it to be similar to other mammalian collagenases. PMID- 3026398 TI - Possible participation of calpain in myosin light chain phosphorylation of human platelets. AB - We previously demonstrated that myosin light chain kinase (MLCK) of gizzard is proteolyzed by platelet calpain. It has been also reported that partially cleaved MLCK may phosphorylate myosin light chain (20K) in the absence of calmodulin. Therefore, a possible participation of calpain in 20K phosphorylation was studied in human platelets, utilizing various inhibitors. An epoxy succinate derivative (E-64) or N-ethylmaleimide (NEM), used as calpain antagonist, inhibited 20K phosphorylation of Ca2+-stimulated lysed platelets. A synergistic effect between these calpain antagonists and calmodulin antagonist W-7 was observed. Also, the similar results were obtained in 20K phosphorylation of intact platelets. From these observations, it was suggested that 20K phosphorylation in platelets is mediated by two separate pathways, namely calmodulin and calpain dependent pathways, provided that calpain activity is specifically inhibited by the antagonists used. PMID- 3026399 TI - Regulation of cytoplasmic poly(A) polymerase activity by dihydrotestosterone in the kidney of rat. AB - Polyadenylation of mRNA, a post-transcriptional phenomenon, is catalysed by cytoplasmic and nuclear poly(A) polymerases. During normal growth and development cytoplasmic poly(A) polymerase activity in the kidney of rat increased from day 20 reaching to a peak at 35 days. Dihydrotestosterone at a dose of 500 micrograms and above significantly stimulated the enzyme activity in the cortex of kidney in the castrated animals. A single dose of dihydrotestosterone caused significant increase in the enzyme activity at 12 h after treatment and was maintained till 24 h. Antiandrogen cyproterone acetate inhibited dihydrotestosterone-induced enzyme activity in kidney cortex. These results show that cytoplasmic poly(A) polymerase of rat kidney is regulated by dihydrotestosterone. PMID- 3026400 TI - Life cycles of cardiac alpha 1- and beta-adrenergic receptors. PMID- 3026401 TI - Protein phosphorylation in rat mast cell granules. Cyclic AMP dependent phosphorylation of a 44K protein associated with broken granules. AB - When rat mast cells are prelabeled with 32PO4 and exposed to non-immunologic or immunologic stimuli under conditions that lead to mediator release from granules, they show rapid increases in labeling of a number of high molecular weight proteins. To determine if granule membrane proteins are subject to protein phosphorylation and perhaps participate in this response, granules with intact or broken membranes were isolated from sonicated, purified rat serosal mast cells on a Percoll gradient. When the granules with broken membranes were incubated with [gamma-32P]ATP and Mg2+ in the absence of exogenous protein kinases, one major radioactive band was recovered in the 44K area after electrophoresis in sodium dodecyl sulfate/polyacrylamide gels. The phosphorylation reaction with ATP required Mg2+, was enhanced by 0.05 to 0.5 microM cyclic AMP, and was inhibited by Ca2+ (0.5 mM and higher). The initial reaction was rapid, and the maximal response was seen at 30 degrees. The 44K band was absent in granules with intact membranes incubated with [gamma-32P]ATP but present when intact granules were lysed with distilled water before adding the [gamma-32P]ATP. These observations indicate that granules have an endogenous phosphorylating system and that the phosphorylation response is on the inner surface of the granule membranes. The possibility was not excluded that a portion of the phosphorylating activity was derived from the cytosol and became firmly associated with broken granules when the intact cells were sonicated. Analysis for possible phosphorylated amino acids in the 44K band after acid hydrolysis showed both phosphoserine and phosphothreonine, indicating that the radioactivity was in a phosphorylated protein or glycoprotein. The 44K phosphorylated protein was made up of several components ranging in pI from approximately 7.6 to 6.6. While the identity of the phosphorylated 44K polypeptide is uncertain, one important possibility is that it is part of an autophosphorylated cAMP dependent protein kinase. The cyclic AMP dependent phosphorylating activity present in granules provides a mechanism by which the granules might respond rapidly to cyclic AMP during mediator release. PMID- 3026403 TI - Bradykinin potentiating peptides isolated from alpha-casein tryptic hydrolysate. PMID- 3026402 TI - Beta-adrenoceptor sensitivity of brown and white adipocytes after chronic pretreatment of rats with reserpine. AB - The effect of pretreatment with reserpine (1.0 mg/kg i.p. daily for 7 days) on beta-adrenoceptor-mediated responses has been measured in epididymal white and interscapular brown adipocytes, left atria and vas deferens of rats in order to investigate the classification of the receptors and whether they are innervated. Lipolysis was measured in adipocytes, and from the same rats, the beta 1 adrenoceptor-mediated positive inotropic responses of isolated paced left atria and beta 2-adrenoceptor-mediated inhibition of field stimulation-induced contraction of the vas deferens were examined. The agonists used were isoprenaline, oxyfedrine (atria only) and (except in brown adipocytes) ritodrine, which was a partial agonist in white adipocytes, atria and vas deferens. Atria and brown adipocytes exhibited beta-adrenoceptor supersensitivity after reserpine pretreatment, whereas vas deferens and white adipocytes did not. Reserpine induced reductions in food intake and body weight did not appear to influence beta-adrenoceptor-mediated lipolysis, since restriction of the diet equivalent to that of reserpine-treated rats produced no change in white adipocyte sensitivity. Responses mediated via beta 1-, but not beta 2-adrenoceptors, display supersensitivity after chronic depletion of neuronal catecholamines with reserpine and this is evidence for innervation of this receptor subtype. Thus, atrial beta 1-adrenoceptors are assumed to be innervated, whereas vas deferens beta 2-adrenoceptors are not. The present results are consistent with histochemical evidence that brown, but not white, adipocyte beta-adrenoceptors are innervated. However, they are not compatible with conventional receptor classification studies, which suggest that rat brown and white beta-adrenoceptors are similar--either both beta 1 or both atypical. PMID- 3026404 TI - Modifications of phospholipid metabolism induced by chlorpromazine, desmethylimipramine and propranolol in C6 glioma cells. AB - The effects of chlorpromazine (CPZ), desmethylimipramine (DMI) and propranolol (PRO) on phospholipid metabolism in C6 glioma cells were studied by following the incorporation of 32Pi, [U-14C]glycerol, [2-3H]glycerol and [1-14C]oleate into lipids. The drugs produced a dose-dependent increase in the incorporation of 32Pi and [U-14C]glycerol, but not of [1-14C] oleate, into total phospholipids, that reached a plateau at 200 microM CPZ and 500 microM DMI and PRO. The three drugs shifted the incorporation of precursors from neutral [phosphatidylcholine (PC) and phosphatidylethanolamine (PE)] to acidic phospholipids [phosphatidic acid (PA), phosphatidylinositol (PI), phosphatidylglycerol, phosphatidylinositol-4 phosphate (PIP) and phosphatidylinositol-4,5-bisphosphate (PIP2)] in a dose dependent, qualitatively similar manner. The incorporation of [2-3H]glycerol into diacylglycerol was also depressed markedly by CPZ. Addition of 1 mM 1,2 dioleoylglycerol, 1-oleoyl-2-acetylglycerol or oleate only partially reversed the decrease in PC labeling caused by CPZ. 12-O-Tetradecanoylphorbol-13-acetate counteracted this effect of CPZ completely but greatly increased PC labeling even in the absence of the drug. Polyphosphoinositides rapidly incorporated 32Pi at early times reaching a plateau in about 40 min. The labeling rate of PI was not parallel to that of PIP or PIP2 and continued to increase even after the polyphosphoinositides had reached a plateau. CPZ increased PI labeling much more than that of PIP and PIP2. These data suggest that cationic amphiphilic drugs may act by inhibiting CTP:phosphocholine cytidylyltransferase, thus decreasing incorporation of precursors into PC and PE; inhibiting PA phosphohydrolase with increased formation of phosphatidyl-CMP, the intermediate for the synthesis of acidic phospholipids; and stimulating the inositol exchange reaction, forming a pool of PI that is not available for PIP and PIP2 synthesis. PMID- 3026405 TI - Altered regulation of adrenal steroidogenesis in 2,3,7,8-tetrachlorodibenzo-p dioxin-treated rats. AB - A single treatment of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) (50 micrograms/kg) produced two distinct effects on adrenal steroidogenesis in rats 13 days post-treatment. In unstressed rats, the very low corticosterone levels early in the light phase (AM) increased 4-fold relative to ad libitum-fed control (ALC) rats, but the peak level of corticosterone that is seen late in the light phase (PM) decreased up to 40% relative to ALC rats. The AM stimulation was also observed in rats pair-fed to compensate for the diminished feed intake of TCDD treated animals, indicating that the change results from nutritional deprivation. The PM suppression, however, was not observed in pair-fed rats. In rats given a lower dose of TCDD (15 micrograms/kg), there was no AM stimulation, whereas the suppression of the PM diurnal peak of corticosterone was retained. Plasma adrenocorticotropin (ACTH) levels and adrenal size were not changed by these treatments, indicating that TCDD affects adrenal responsiveness. TCDD did not, however, have a significant effect on corticosterone secretion in rats receiving high doses of ACTH. In control animals, the availability of cholesterol to cytochrome P-450scc limits the rate of steroidogenesis. While the specific content of the cytochrome was unaffected by TCDD, cholesterol turnover by this enzyme appeared to be affected following TCDD treatment, as evidenced by small increases in the mitochondrial levels of free cholesterol, reactive cholesterol, and in the proportion of P-450scc complexed with cholesterol relative to both ad libitum- and pair-fed controls. This accumulation of mitochondrial cholesterol following TCDD treatment is consistent with an inhibition of cholesterol metabolism at cytochrome P-450scc in vivo that is removed upon isolation of the mitochondria. These TCDD-induced increases were enhanced substantially in ACTH stimulated rats, probably because ACTH enhances cholesterol influx into the mitochondria. Normally, substrate availability is rate limiting in cholesterol side-chain cleavage, and the AM stimulation of steroidogenesis by TCDD may result from such increased cholesterol transfer. The inhibition of cholesterol side chain cleavage resulting from TCDD treatment may, however, only become rate limiting for corticosterone synthesis when cholesterol transfer is more substantially activated, as for peak PM secretion. PMID- 3026406 TI - High sodium diet and Na+-stimulated ATPase activities in basolateral plasma membranes from rat kidney proximal tubular cells. AB - The ouabain-insensitive, Na+-stimulated ATPase activity of kidney proximal tubular cells from rats fed a high Na+ diet for 4 months was increased approximately 70% when compared with control (normal diet) rats. The higher ATPase activity was not due to a change in the affinity of the system toward ATP, Mg2+ or Na+. This increase in Na+-ATPase activity may be due to either a higher number of pumps or to a higher turnover rate of the enzyme or both. The ouabain sensitive, Na+, K+-stimulated ATPase activity, on the other hand, did not change with the high sodium diet. These results can be taken as evidence that the Na+,K+ ATPase and the Na+-ATPase of basolateral plasma membranes of proximal tubular cells from rat kidney are two different entities. PMID- 3026407 TI - Studies on the mechanism of action of the gastric microsomal (H+ + K+)-ATPase inhibitors SCH 32651 and SCH 28080. AB - The novel antisecretory agents SCH 32651 and SCH 28080 were evaluated for their antisecretory activities in vitro as well as for their abilities to inhibit the (H+ + K+)-ATPase enzyme activity in preparations of microsomal membranes from rabbit fundic mucosa. SCH 32651 and SCH 28080 inhibited both the histamine- and dibutyryl cAMP-stimulated uptake of [14C]-aminopyrine into isolated parietal cells with IC50 values of about 1.5 and 0.02 microM respectively. SCH 32651 and SCH 28080 competitively inhibited the K+-stimulated hydrolysis of ATP catalyzed by the (H+ + K+)-ATPase with Ki values of 16.3 and 0.12 microM respectively. The inhibition of the enzyme by both compounds was not affected by the addition of the sulfhydryl reducing agents dithiothreitol or beta-mercaptoethanol, was readily reversible by dilution or washing, and was dependent upon the concentration of KCl used to stimulate the enzyme. These data suggest that SCH 32651 and SCH 28080 are reversible, competitive inhibitors of the K+-stimulated hydrolysis of ATP. PMID- 3026408 TI - [Complex formation of single-stranded phage DNA with synthetic oligodeoxynucleotides and interaction of complexes with DNA-polymerase and restriction endonuclease]. AB - Hybridization of synthetic oligodeoxynucleotides with single-stranded phage M13mp2 DNA has been studied in terms of temperature, ionic strength, oligonucleotide molar excess and chain length, and DNA secondary structure. Combination of two decadeoxynucleotides corresponding to a nicked eicosamer (composite primer) was found to be efficient in the template-directed DNA polymerase-catalyzed chain elongation, where both decamers separately failed. Circular SS DNA was specifically linearized by BamHI cleavage of SS DNA tetradecadeoxynucleotide duplex. PMID- 3026409 TI - Products of polymorphonuclear cell injury inhibit IgG enhancement of monosodium urate-induced superoxide production. AB - The generation of polymorphonuclear cell (PMN) superoxide ion (O2-) by monosodium urate (MSU) crystals may be important in the pathogenesis of acute gout. Coating MSU crystals with IgG prior to exposure to PMN markedly augmented O2- generation. This augmentation was inhibited by supernates, termed cell lysate, derived from sonicated PMN or PMN exposed to MSU crystals for 5 hours at 37 degrees C. Lysate was effective in inhibiting O2- production when incubated with MSU crystals prior to, during, or after MSU crystals were exposed to IgG. No IgG could be eluted from crystals exposed to both lysate and IgG. Immunoelectron microscopy showed virtually no IgG on crystal surfaces after incubation of crystals with lysate and IgG. These data suggest that products of PMN injury can modulate further PMN responses to MSU crystals. This phenomenon provides a negative feedback loop and is one possible mechanism for the self-limitation of acute gouty attacks. PMID- 3026410 TI - Inorganic pyrophosphate release by rabbit articular chondrocytes in vitro. AB - Release of inorganic pyrophosphate (PPi) by rabbit articular chondrocytes in vitro was measured by a newly developed assay which utilizes radioactive orthophosphate (32Pi) labeling and anion exchange high performance liquid chromatography. Chondrocytes in monolayer and high density culture failed to release PPi. Explants (cartilage fragments), however, released newly formed PPi into the culture medium. Trypsin treatment of cartilage fragments almost completely blocked the PPi extrusion. Collagenase treatment had no effect on PPi extrusion. There was no clear correlation between proteoglycan synthesis, measured by 35SO4 incorporation, and PPi release. Suppression of proteoglycan synthesis with tunicamycin did not influence the PPi release of the explants. PMID- 3026412 TI - Long-term effect of mackerel diet on blood pressure, serum lipids and thromboxane formation in patients with mild essential hypertension. AB - Twelve male patients with mild essential hypertension were put on a diet supplemented with 2 cans of mackerel/day (= 2.2 g daily of eicosapentaenoic acid, EPA, C20:5 n-3 and 2.8 g daily of docosahexaenoic acid, DHA, C22:6 n-3) for 2 weeks within an isocaloric regimen and then with 3 cans/week (= 3.3 g/week, equivalent to 0.47 g daily of EPA and 4.2 g/week, equivalent to 0.69 g daily of DHA) for 8 months with a subsequent period of 2 months on normal diet. Eleven male hypertensives matched for age, body weight index, blood pressure and serum lipids with no change in their nutritional habits served as controls. After the first dietary period (2 weeks) a significant decrease of serum triglycerides (TG), total and LDL-cholesterol, blood pressure and thromboxane B2 (TxB2) was found, whereas HDL cholesterol and potassium in erythrocytes were significantly increased. During the second dietary period (8 months) providing the lower dose of EPA, serum lipids and the other biochemical parameters returned to the initial values. Blood pressure, however, remained significantly lower and rose to the basal levels only after the third period (2 months) on normal diet. In the control group no alterations could be seen. The data suggest a dose-related differential effect of dietary EPA on serum lipids, lipoproteins, TxB2 and blood pressure in subjects with mild hypertension. PMID- 3026411 TI - Soy fiber improves lipid and carbohydrate metabolism in primary hyperlipidemic subjects. AB - This study was designed to evaluate the effects of soy fiber, a natural source of dietary fiber that consists of both cellulosic and noncellulosic dietary fiber, on human plasma lipoprotein lipids and glucose tolerance in patients with primary hyperlipidemia. Supplementing 25 g of soy fiber per day provided a significant additional reduction of plasma total-cholesterol by 13 mg/dl (P less than 0.04) and LDL cholesterol by 12 mg/dl (P less than 0.05) beyond that previously achieved by treatment with an NIH Type II-A low-fat, low-cholesterol diet for 12 weeks in Type II-A hypercholesterolemic patients. There were no effects on HDL cholesterol or apoprotein A-I and A-II levels. The hypocholesterolemic effect was greater than in the hyperlipidemic patients with impaired glucose tolerance. Soy fiber supplementation also significantly reduced insulin responses to oral glucose challenge by 20% in Type II-A hypercholesterolemic and by 16.5% in Type IV hypertriglyceridemic patients. Results from this study suggest that supplementing the diet with soy fiber may be beneficial in dietary management of hyperlipidemia in patients with hypercholesterolemia and particularly in hyperlipidemic patients with hyperinsulinemia and impaired glucose tolerance. PMID- 3026413 TI - Late onset congenital adrenal hyperplasia in a Puerto Rican family: hormonal and HLA typing. PMID- 3026414 TI - Intravenous drug abuse and the accident and emergency department: AIDS (HTLV-III) antibodies and hepatitis B markers. PMID- 3026415 TI - [Pipercuronium bromide (Arduan)--a new non-depolarizing muscle relaxant]. PMID- 3026416 TI - [Nursing follow-up after hospital discharge]. PMID- 3026417 TI - [Mixed rapid restorative technics]. PMID- 3026418 TI - [Anti-ulcer action and peripheral opioid activity of the products of dalargin degradation]. PMID- 3026419 TI - Cholera toxin as an intracellular cAMP-inducing agent in the regulation of haemopoietic cell development and differentiation. PMID- 3026420 TI - Cloning and in vitro expression of the measles virus matrix gene. AB - A cDNA library was prepared from Vero cells infected with the Edmonston strain of measles virus. A number of viral specific cDNA clones were isolated and characterized by Northern blot hybridization analysis. A cDNA clone containing a 1500 base pair insert which hybridizes to a viral specific transcript of approximately 1500 nucleotides was subcloned into pSP64 and used as an in vitro transcription template. The resulting RNA transcript was translated in a cell free system, giving rise to a polypeptide which comigrates on polyacrylamide gels with the authentic measles virus matrix protein and is immunoprecipitated with antisera specific for the matrix protein. PMID- 3026422 TI - Diacylglycerol lipase and kinase activities in rabbit aorta and coronary microvessels. AB - Diacylglycerol lipase and kinase activities were measured in particulate and soluble fractions from rabbit aorta (intima-media) and coronary microvessels. With rabbit aorta, the hydrolysis at the sn-1 position of 1-palmitoyl-2-oleoyl-sn glycerol had a pH optimum of 5-6 and was greater than hydrolysis at the sn-2 position (pH optimum of 6.5). Only the 2-monoacylglycerol accumulated during incubations at pH 5 and 6.5. These results are consistent with an ordered two step reaction sequence where the fatty acid at the sn-1 position is released first, followed by the hydrolysis of the fatty acid from the 2-monoacylglycerol by a monoacylglycerol lipase with a neutral pH optimum. Lipase activity (sn-2 hydrolysis) at pH 6.5 was greater than kinase activity at all substrate concentrations. The presence of arachidonate at the sn-2 position of the diacylglycerol increased kinase activity but had little effect on lipase activity. Kinase activity was mainly particulate, whereas 50-60% of diacylglycerol lipase and 50% of monoacylglycerol lipase activity were soluble. Diacylglycerol lipase and kinase were also present in coronary microvessel preparations. Diacylglycerol lipase (sn-2 hydrolysis) activity in coronary microvessels was not enhanced by preincubation of the enzyme preparation with cAMP-dependent protein kinase. PMID- 3026421 TI - Digitalis-like biological activity in rat cerebellum. AB - The cytosolic fraction of rat cerebellum possesses a factor(s) which is capable of inhibiting synaptosomal Na,K-ATPase activity, competing with [3H]ouabain binding to rat brain synaptosomes, and inducing positive inotropy in guinea pig atrial strips. These results demonstrate the existence of a ouabain-like principle in rat cerebella. The inhibitory activity of the factor was found to be partially thermolabile and diminished by a proteolytic agent, and the activity could be augmented by increasing concentrations of Mg2+, suggesting a regulatory mechanism for the endogenous digitalis-like principle. PMID- 3026423 TI - Purification of antigens of mammary tumor of the mouse by gel filtration. AB - This study was designed to isolate the biologically active components from a perchloric acid (PCA) extract of mammary tumor virus (MTV)-induced mouse mammary tumors (AMMT) previously shown to be capable of inducing DNA synthesis in spleen cells of mice presensitized by inoculation of viable syngeneic mammary tumor cells. AMMT (450 mg) was eluted fom an Ultrogel AcA54 column with separation range of 5-70K in a buffer containing Tris-sodium chloride and EDTA. Two fractions were separated with estimated molecular weights of 68-70K (AMMT I) and 35-40K (AMMT II). AMMT I and AMMT II exhibited DNA synthetic activity 78-fold and 35-fold, respectively, greater than AMMT. The feasibility of further purification of AMMT I by isoelectric focusing was also demonstrated. These components provide the opportunity for further investigation into the molecular basis of tumor-host relationship in this animal model. PMID- 3026424 TI - The effects of aluminum loading on the renal response to parathyroid hormone in the vitamin D-replete rat. AB - Aluminum (Al) may cause vitamin D-resistant osteomalacia and depress the serum levels of immunoreactive parathyroid hormone (iPTH) in patients treated with maintenance dialysis and those on total parental nutrition (TPN). Both conditions have been associated with low serum levels of 1,25(OH)2-vitamin D (1,25(OH)2D). Al may inhibit PTH secretion in vitro; however, induction of hypocalcemia can enhance endogenous PTH secretion in Al-loaded dogs and TPN patients. Despite hypocalcemia and/or increased endogenous iPTH levels, Al-loaded TPN patients fail to show the expected rise in serum 1,25(OH)2D levels. Such observations suggest that Al may impair the renal response to PTH. We studied vitamin D-replete rats given Al or saline vehicle IP for 5 days. Al and control rats then received a saline infusion with an IV bolus of PTH 1-34. Urinary cyclic AMP and P excretion rose in Al and control rats by 1 hr post-PTH, without differences between the groups. Serum P and ionized Ca levels were not different between Al and control rats. In other Al and control rats, serum 1,25(OH)2D levels were measured after saline without PTH. Serum 1,25(OH)2D levels were higher in controls given PTH than in those without, but 1,25(OH)2D levels were not different between Al rats given PTH and those with none. Thus, aluminum does not affect cyclic AMP or P excretion but may impair 25(OH)D-1 alpha-hydroxylase activity in response to PTH. PMID- 3026425 TI - Patterns of reactivity of the monoclonal antibody 791T/36 with different tumour metastases in the liver. AB - Reactivity of the monoclonal antibody 791T/36 with secondary malignant deposits has been investigated in frozen sections of 74 human liver biopsy specimens. There was no reactivity with hepatocytes but in some instances binding to fibrous tissues and in one case to portal tract lymphocytes was observed. Sections from 9 biopsy specimens contained malignant deposits. In seven of these 791T/36 bound either to malignant cells or to pseudoacinar contents (3 colorectal adenocarcinomas; 1 probable pancreatic adenocarcinoma; 1 medullary cell carcinoma of thyroid; 1 oat cell carcinoma of bronchus and 1 deposit of nodular sclerosing Hodgkins Disease). Two undifferentiated tumours (1 gastric adenocarcinoma and 1 oat cell bronchial carcinoma) showed no antibody binding. The histological pattern of reactivity previously reported with primary tumours appears to be similar in secondary deposits. A wider range of tumours than recognised hitherto binds 791T/36. Whether the binding to fibrous tissue seen in some instances is sufficient to cause diagnostic uncertainty when 791T/36 is used for scanning requires further investigation. PMID- 3026426 TI - Mortality of hepatoma and cirrhosis of liver in Taiwan. AB - A study of mortality from hepatoma and hepatic cirrhosis was conducted in Taiwan, where their mortality rates are among the highest in the world in 1980 being 26.10 and 8.14 per 100,000 population for males and females, respectively, for hepatoma, and 33.01 and 12.90 for males and females, respectively, for cirrhosis. The secular trends of hepatoma and hepatic cirrhosis death rates have been increasing, especially in males, with consequent increase in the sex ratio. The large difference in mortality rates between males and females and the increasing trends in the sex ratio suggest that other factors besides hepatitis B virus (HBV), are involved in the aetiology of hepatoma and cirrhosis of liver. PMID- 3026427 TI - Determinants of pulmonary fibrosis and lipidosis in the silica model. AB - The conditions which might favour development of the fibrotic or the lipid component of the pulmonary reaction to inhaled quartz were examined in rats. Smaller particle size and freedom from surface contamination by amorphous silica or iron oxide, status of the animals whether specific pathogen-free or conventional, and the resistance of cell membranes to damage appeared to bear on fibrogenesis. Increased membrane stability by treatment with polyvinylpyridine-N oxide abolished not only the fibrosis but also the response of type II cells and hence lipidosis. The rate and intensity of quartz deposition may also affect the response, a low concentration inhaled over a long period favouring nodulation. No other manipulations, environmental or pharmacological, succeeded in inhibiting lipidosis to the benefit of fibrosis. Guinea pigs, however, behaved differently, their reaction being characterized by massive alveolar accumulation of dust bearing macrophages and type II cell hyperplasia but not by lipidosis. The species variation is unexplained but macrophage predominance may represent a phase that later transforms to lipidosis. The experimental findings may have implications for forms of pneumoconiosis other than silicosis. PMID- 3026428 TI - The development of mercury- and selenium-containing deposits in the kidneys following implantation of dental amalgams in guinea pigs. AB - Examination of light microscopical sections of the kidneys of guinea pigs with chronic exposure to mercury as the result of the breakdown of subcutaneous implants of powdered dental amalgam demonstrated the development of black, refractile deposits in the cytoplasm and nuclei of cells in both the straight and convoluted portions of the proximal tubule. The more numerous cytoplasmic deposits were of a particulate nature with dimensions of approximately 1 microgram. The nuclear deposits, which appeared later but which were relatively more common in longer-term animals, took the form of prominent inclusions, 1 to 3 micrograms in diameter. The ratio of nuclear to cytoplasmic deposits was higher in animals receiving high copper as compared with conventional amalgam. At electron microscopical level, the cytoplasmic deposits were seen to consist of collections of fine particles within lysosomes. Similar deposits were also found in far smaller numbers in lysosomes in collecting duct cells. The nuclear inclusions in proximal tubular cells were made up of closely packed electron dense granules. X-ray microanalysis showed both lysosomal and nuclear deposits to contain mercury and selenium. The association of mercury with selenium, which was present in the animals' diet at low levels, probably aided the microscopical visualisation of the deposits. PMID- 3026429 TI - Langerhans cells in human warts. AB - Seventy-six warts (15 plantar, 38 hand, 16 miscellaneous and seven anogenital lesions) taken from 55 patients, were studied by indirect immunofluorescence with monoclonal antibodies specific for T-cell subsets, Langerhans cells (LC) and HLA DR antigen. The results were related to the presence of viral antigen. Approximately 80% of the lesions showed an infiltrate. Only 19 lesions contained helper/inducer or suppressor/cytotoxic T cells. The distribution of LC was abnormal in 65% of biopsies which contained LC in the dermis, and 29% were devoid of LC in the epidermis. Many lesions had reduced numbers of LC in the epidermis. The disappearance of LC from the epidermis was related to the presence of viral antigen, but not to the presence of particular T-cell subsets. Infiltrating cells were sometimes HLA-DR-positive, whereas basal cells did not express HLA-DR antigen, irrespective of the density of the infiltrate. PMID- 3026430 TI - The effect of thalidomide on arachidonic acid metabolism in human polymorphonuclear leukocytes and platelets. AB - The effect of thalidomide on the metabolism of arachidonic acid by purified washed human platelets and polymorphonuclear leukocytes was examined. The drug was found to be without effect under the conditions used. The action of thalidomide cannot therefore be attributed to a direct inhibition of prostaglandin or leukotriene biosynthesis. PMID- 3026431 TI - Human papillomavirus infections in a group of renal transplant recipients. AB - One hundred and twenty renal transplant recipients were investigated. Fifty-eight (48%) were found to have warts, 13 (11%) keratoses and six (5%) to have, or recently to have had cancers. The longer the time of immunosuppression, the greater the prevalence of warts; of those patients who had had their transplant for at least 5 years, 87% had warts. Those with a graft survival time of 10 years or more are at special risk of warts, keratoses and malignancy. Five (10%) of 50 women had genital warts, four of whom had internal lesions (vaginal, cervical or anal) and one developed a carcinoma of the vulva. These findings indicate the advisability of colposcopy for all female renal transplant recipients, a high risk group. Eighty-eight specimens from 42 patients were examined by DNA restriction enzyme analysis and cross hybridization for the presence and type of human papillomavirus (HPV). HPV DNA was detected in 66% of the warts examined, HPV2 and HPV4 occurring most often and HPV1 and HPV3 only infrequently. In sequential specimens from common hand warts of one individual, an HPV was found which could not be precisely identified but was related to HPV4. HPV16 was detected in a vaginal wart from one patient and an HPV6-related virus in a vulval wart of another. HPV DNA of an unknown type was demonstrated in one of 11 keratoses examined. With the probes used to examine the few samples of skin cancers available, HPV16 was found in a squamous cell carcinoma of the vulva, and faint bands from an unidentified type of HPV were detected in two squamous cell carcinomata from a patient's hand. One woman had plaque lesions morphologically and histologically resembling those found in epidermodysplasia verruciformis (EV). HPV5 was identified in these lesions. This is only the third reported case of HPV5, previously thought to be unique to EV, in a renal transplant recipient. PMID- 3026432 TI - Interferon responses of peripheral blood mononuclear cells from normal and leukaemic children. AB - Human peripheral blood mononuclear cell cultures (PBMC) stimulated with Sendai virus or K562 cells produce a mixture of interferons. Temperature and pH stability characteristics and reactions with monospecific antibodies indicate that PBMC cultures from adults produce interferons alpha and gamma in approximately equal proportions. PBMC cultures from children produce lower levels of interferons with a higher proportion of type alpha. The ability of PBMC cultures from children with acute lymphoblastic leukaemia (ALL) to produce interferon was determined. Little or no gamma interferon was induced by either Sendai virus or K562 cells. Cultures from some children with ALL produced alpha interferon but mean levels were significantly lower than from normal children. A group of older children with ALL who had completed their course of therapy and were off treatment produced levels of interferon indistinguishable from those of normal children. This in vitro deficit, possibly induced by chemotherapy, may reflect an in vivo deficit and may contribute to the impaired handling of viruses seen in children being treated for ALL. PMID- 3026433 TI - Certain myeloid cells possess receptors for interleukin-2. AB - A proportion of blasts from five of 10 cases of AML expressed receptors for IL-2 (IL-2R) when tested directly ex-vivo with monoclonal antibodies against the receptor. After in-vitro stimulation with various agents including TPA, gamma interferon and colony stimulating factor, the purified blast cells of all cases of AML tested (10 of 10) showed high levels (50-90% cells positive) of IL-2R expression. Granulocytic cells from the promyelocyte stage onwards lacked IL-2R both before and after in-vitro stimulation. In contrast, leukaemic promonocytes and normal peripheral monocytes expressed IL-2R both before and after stimulation. The receptors were detected with two different monoclonal antibodies (anti-Tac and 4H3--both IgG 2a antibodies) by indirect rosetting. In selected experiments, results were confirmed by fluorescence microscopy and by the APAAP technique. Normal monocytes possessed only small amounts of IL-2R since positivity was clearly detectable only by the indirect rosette assay. Irrelevant IgG 2a first-layer and second-layer-only controls were always negative. The endogenous nature of the IL-2 receptors was demonstrated by re-expression after capping and shedding. That the antigens detected were true IL-2R was confirmed by the fact that monoclonal antibody staining was blocked by recombinant IL-2. The restricted expression of IL-2R on early granulocytic and on monocytoid cells raises the possibility that IL-2 is important in the proliferation of these cell types. PMID- 3026434 TI - Phenotypic characterisation of peripheral blood lymphoid cells in people exposed to fibrous zeolite. AB - Among inhabitants of the village of Karain in Turkey there is an extremely high incidence of malignant mesothelioma, most probably due to exposure to erionite, which is a fibrous zeolite and similar in appearance and properties to asbestos. This mineral may be found in the dust in the village. To characterise possible disturbances in the immune system of people exposed to fibrous zeolite, a phenotypic characterisation of lymphoid cells in the peripheral blood of 74 immigrants to Sweden from Karain was performed. Compared with normal controls, the mean percentages of Leu 4+ cells (Pan-T) and Leu 3a+ cells ("helper/inducer" T cells) were significantly decreased, whereas the mean percentage of Leu 2a+ cells ("suppressor/cytotoxic" T cells) was normal, leading to a significant reduction of the Leu 3a/Leu 2a subset ratio. The percentage of B cells (Leu 12+ cells) was significantly increased, whereas the percentages of both HLA-DR+ and HLA-DQ+ cells were normal. The percentage of natural killer cells (NK) and killer (K) cells as defined by the monoclonal anti-Leu 7 and anti-Leu 11b were also normal. These findings indicate that exposure to fibrous zeolite causes a numerical imbalance between the two phenotypically different T cell subsets similar to that seen in asbestos exposed individuals. PMID- 3026435 TI - A simplified method for determining erythrocyte pyrimidine 5'-nucleotidase (P5N) activity by HPLC and its value in monitoring lead exposure. AB - The method for determining erythrocyte pyrimidine 5'-nucleotidase (P5N EC 3.1.3.5) activity has been simplified using an automated high performance liquid chromatograph (HPLC). The activity determined by the simplified method agreed closely with that obtained by conventional methods. In 161 lead workers P5N activity declined linearly with increasing blood lead concentrations (Pb-B) between 20 and 80 micrograms/100 g, and correlated well with Pb-B (r = -0.87). For the same group of workers, correlation coefficients between Pb-B v ALA-D activity, zinc protoporphyrin, ALA-U, and coproporphyrin were -0.87, 0.73, 0.70, and 0.32, respectively. At Pb-B greater than or equal to 40 micrograms/100 g, the validity of P5N (1.86 at a cut off of 10 less than or equal to units) was higher than that of other indicators examined. P5N activity was fairly stable during the storage of samples for two weeks at 4 degrees C. Determination of P5N activity by this method may be a useful indicator in screening for moderate exposure to lead. PMID- 3026436 TI - Comparative effects of dietary wheat bran and its morphological components (aleurone and pericarp-seed coat) on volatile fatty acid concentrations in the rat. AB - Adult male rats were fed on diets containing 100 g dietary fibre/kg either as alpha-cellulose or wheat bran or the pericarp-seed coat or aleurone layers prepared from that bran by sequential milling and air elutriation and electrostatic separation. After 10 d, concentrations of total volatile fatty acids (VFA) in caecal fluid were significantly different between groups and fell in the order: aleurone greater than wheat bran greater than pericarp-seed coat greater than cellulose. This ranking probably reflected the ease of fermentation of fibre polysaccharides by colonic bacteria which also resulted in a considerably higher faecal bacterial mass in the aleurone group. Because of the differences in the volume of caecal digesta, the mass of caecal VFA was considerably the highest in the aleurone group, intermediate with wheat bran and equally low in the pericarp-seed coat and cellulose groups. The diet based on aleurone gave a relatively higher proportion of propionate but with both pericarp seed coat and wheat bran the contribution of butyrate was raised. VFA concentrations in hepatic portal venous plasma were proportional to caecal concentrations with very high (greater than 3 mM) values being recorded in the aleurone group. The findings are discussed in relation to the apparent susceptibility of the morphological components of wheat bran to fermentation by large bowel bacteria. PMID- 3026437 TI - Amino acid sequence of the a subunit of human factor XIII. AB - Factor XIII is a plasma protein that plays an important role in the final stages of blood coagulation and fibrinolysis. The complete amino acid sequence of the a subunit of human factor XIII was determined by a combination of cDNA cloning and amino acid sequence analysis. A lambda gtll cDNA library prepared from human placenta mRNA was screened with an affinity-purified antibody against the a subunit of human factor XIII and then with a synthetic oligonucleotide probe that coded for a portion of the amino acid sequence present in the activation peptide of the a subunit. Six positive clones were identified and shown to code for the a subunit of factor XIII by DNA sequence analysis. A total of 3831 base pairs was determined by sequencing six overlapping cDNA clones. This DNA sequence contains a 5' noncoding region or a region coding for a portion of a pro-piece or leader sequence, the mature protein (731 amino acids), a stop codon (TGA), a 3' noncoding region (1535 nucleotides), and a poly(A) tail (10 nucleotides). When the a subunit of human factor XIII was digested with cyanogen bromide, 11 peptides were isolated by gel filtration and reverse-phase HPLC. Amino acid sequence analyses of these peptides were performed with an automated sequenator, and 363 amino acid residues were identified. These amino acid sequences were in complete agreement with those predicted from the cDNA. The a subunit of factor XIII contained the active site sequence of Tyr-Gly-Gln-Cys-Trp, which is identical with that of tissue transglutaminase.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3026438 TI - Preferred sites on cytochrome c for electron transfer with two positively charged blue copper proteins, Anabaena variabilis plastocyanin and stellacyanin. AB - Rate constants for the reactions of horse cytochrome c (E'0 of +260 mV) with the copper proteins Anabaena variabilis plastocyanin (E'0 of +360 mV) used as oxidant and stellacyanin (E'0 of +187 mV) used as reductant have been determined at 25 degrees C, pH 7.5 and 7.0, respectively, and an ionic strength of 0.10 M (NaCl). These rate constants were also measured with eight different singly substituted 4 carboxy-2,6-dinitrophenyl (CDNP) horse cytochrome c derivatives, modified at lysine-7, -13, -25, -27, -60, -72, -86, or -87 and with the trinitrophenyl (TNP) derivative modified at lysine-13. The influence of the modifications on the bimolecular rate constants for these reactions defines the region on the protein that is involved in the electron-exchange reactions and demonstrates that the preferred site is at or near the solvent-accessible edge of the heme prosthetic group on the "front" surface of the molecule. Both reactions are strongly influenced by the lysine-72 modification to the left of the exposed heme edge and, to this extent, behave similar to the earlier studied reaction with azurin. These effects span only an order of magnitude in rate constants and are thus many times smaller than those for the physiological protein redox partners of cytochrome c. While the preferred sites of reaction on the surface of cytochrome c for small inorganic complexes appear to be dependent only on the net charge of the reactants, with the copper proteins additional factors intervene. These influences are discussed in terms of hydrophobic patches and the distribution of charges on the surface of the four copper proteins so far examined. PMID- 3026440 TI - Folding/unfolding kinetics of mutant forms of iso-1-cytochrome c with replacement of proline-71. AB - Proline-71, an evolutionally conserved residue that separates two short alpha helical regions, is replaced by valine, threonine, or isoleucine in at least partially functional forms of iso-1-cytochrome c from Saccharomyces cerevisiae [Ernst, J. F., Hampsey, D. M., Stewart, J. W., Rackovsky, S., Goldstein, D., & Sherman, F. (1985) J. Biol. Chem. 260, 13225-13236]. To assign the effects of perturbations at position 71 to steps in the process of protein folding, the kinetic properties of the folding/unfolding reactions of normal protein and the three mutant forms are compared. At pH 6.0, 20 degrees C, fluorescence-detected folding/unfolding kinetics are monitored below, within, and above the equilibrium transition zone by using stopped-flow mixing to perform guanidine hydrochloride concentration jumps. Three kinetic phases are detected for each of the four proteins. The fastest of these phases (tau 3) differs in rate for the wild type and mutant proteins. The remaining kinetic phases (tau 1 and tau 2) have similar rates for all four proteins over the entire range of folding/unfolding conditions. The guanidine hydrochloride dependence of the relative amplitudes of the kinetic phases is complex and is sensitive to the nature of the substituent at position 71: each of the four proteins shows differences in the fraction of folding/unfolding associated with the two fastest rate processes. The results suggest that it is the location of the mutation in the primary structure rather than the nature of the substituent that determines which kinetic step (or steps) is changed in rate.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3026439 TI - Guanidine hydrochloride induced equilibrium unfolding of mutant forms of iso-1 cytochrome c with replacement of proline-71. AB - Proline-71, an evolutionally conserved residue that separates two short alpha helical regions, is replaced by valine, threonine, or isoleucine in at least partially functional forms of iso-1-cytochrome c from Saccharomyces cerevisiae [Ernst, J. F., Hampsey, D. M., Stewart, J. W., Rackovsky, S., Goldstein, D., & Sherman, F. (1985) J. Biol. Chem. 260, 13225-13236]. Treatment of these proteins with a specific sulfhydryl blocking reagent (methyl methanethiosulfonate) to block Cys-102 has allowed investigation of the properties of monomeric forms of the proteins, denoted iso-1-MS. Comparison of the UV-visible absorbance properties (pH 6, 20 degrees C) shows minor differences between the normal Pro-71 iso-1-MS and two of the three mutant proteins. The Val-71 iso-1-MS protein has absorbance properties indistinguishable from those of the normal Pro-71 iso-1-MS protein, but the Ile-71 iso-1-MS and Thr-71 iso-1-MS proteins show reduced intensity of the 695-nm absorbance band and a small shift in the Soret maximum, from 408 nm for the Pro-71 iso-1-MS and Val-71 iso-1-MS proteins to 406 nm for the Thr-71 iso-1-MS and Ile-71 iso-1-MS proteins. Second derivative spectroscopy is used to assess differences in the polarity of the environment of tyrosine residues. The average degree of exposure of tyrosines to solvent is similar in all four proteins: 0.39 for the normal Pro-71 iso-1-MS and Val-71 iso-1-MS proteins; 0.40 for the Ile-71 iso-1-MS protein.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3026441 TI - Effect of Al3+ plus F- on the catecholamine-stimulated GTPase activity of purified and reconstituted Gs. AB - The effects of Al3+ and F- on the catecholamine-stimulated GTPase cycle were studied by using reconstituted phospholipid vesicles that contained purified beta adrenergic receptor and the stimulatory GTP-binding protein of the adenylate cyclase system, Gs. Al3+/F- activated reconstituted Gs to levels previously reported for detergent-solubilized, purified Gs, although both activation and deactivation were faster in the reconstituted preparation. Under these conditions, Al3+/F- did not inhibit by more than 15% the beta-adrenergic agonist stimulated GTPase activity of the vesicles nor did it significantly inhibit the rates of GTP binding, GTP hydrolysis, or GDP release. When Mg2+ (50 mM) was used instead of agonist to promote GTP hydrolysis in the receptor-Gs vesicles, Al3+/F- was found to inhibit GTP gamma S binding, GDP release, and steady-state GTPase activity to unstimulated levels. These data can be interpreted as indicating that the receptor catalyzes nucleotide exchange by Gs faster or more efficiently than does Mg2+. PMID- 3026442 TI - Affinity-purified tetanus neurotoxin interaction with synaptic membranes: properties of a protease-sensitive receptor component. AB - The pharmacokinetic interaction of an affinity-purified 125I-labeled tetanotoxin fraction with guinea pig brain synaptosomal preparations was investigated. Binding of tetanotoxin was time- and temperature-dependent, was proportional to protein concentration, and was saturable at about 8 X 10(-9) M as estimated by a solid-surface binding assay. Binding was optimal at pH 6.5 under low ionic strength buffer and was almost entirely blocked by gangliosides or antitoxin. In analogy to intact nerve cells, binding of toxin to membranes resulted in a tight association operationally defined as sequestration. Binding and sequestration were abolished after membrane pretreatment with sialidase. The enzyme could not dissociate the membrane-bound toxin formed at 4 or 37 degrees C under low ionic strength conditions, which is in part compatible with internalization as defined in nerve cell cultures. In the latter system the toxin could be removed at 4 degrees C but not at 37 degrees C. Binding was significantly reduced upon pretreatment of guinea pig brain membranes by a variety of hydrolytic enzymes. Trypsin and chymotrypsin inhibited binding between 55% and 68% while bacterial protease abolished it by 91-95%. The effect was species-specific as it was not seen in rat or bovine synaptosomes. Collagenase and hyaluronidase had little or no inhibitory effect when applied to synaptosomes (27% and 9%) but inhibited binding to synaptic vesicles by 56% and 49%, respectively. Phospholipases A2 and C caused 42-43% inhibition of binding in vesicles and less than 22% in synaptosomes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3026443 TI - Direct incorporation of guanosine 5'-diphosphate into microtubules without guanosine 5'-triphosphate hydrolysis. AB - Using highly purified calf brain tubulin bearing [8-14C]guanosine 5'-diphosphate (GDP) in the exchangeable nucleotide site and heat-treated microtubule-associated proteins (both components containing negligible amounts of nucleoside diphosphate kinase and nonspecific phosphatase activities), we have found that a significant proportion of exchangeable-site GDP in microtubules can be incorporated directly during guanosine 5'-triphosphate (GTP) dependent polymerization of tubulin, without an initial exchange of GDP for GTP and subsequent GTP hydrolysis during assembly. The precise amount of GDP incorporated directly into microtubules is highly dependent on specific reaction conditions, being favored by high tubulin concentrations, low GTP and Mg2+ concentrations, and exogenous GDP in the reaction mixture. Minimum effects were observed with changes in reaction pH or temperature, changes in concentration of microtubule-associated proteins, alteration of the sulfonate buffer, or the presence of a calcium chelator in the reaction mixture. Under conditions most favorable for direct GDP incorporation, about one-third of the GDP in microtubules is incorporated directly (without GTP hydrolysis) and two-thirds is incorporated hydrolytically (as a consequence of GTP hydrolysis). Direct incorporation of GDP occurs in a constant proportion throughout elongation, and the amount of direct incorporation probably reflects the rapid equilibration of GDP and GTP at the exchangeable site that occurs before the onset of assembly. PMID- 3026444 TI - Chromatin structure of the cytochrome P-450c gene changes following induction. AB - The chromatin structure of cytochrome P-450c and P-450d genes, which in the liver are highly inducible by 3-methylcholanthrene, was studied in normal and carcinogen-treated rats by using a cDNA probe specific for P-450c and a genomic probe that recognizes both genes. Digestion with micrococcal nuclease revealed that the active genes are not present in the typical 200 base pair nucleosomal structure. Gene induction is associated with a rearrangement of the nuclear organization of the genes. By use of indirect end-label hybridization, three DNase I hypersensitive sites were mapped, one in the 5'-terminal region and two in the 3' region of the P-450c gene. Gene induction, by treatment with 3 methylcholanthrene, changes the location of the DNase I site present in the 5' region without affecting the sites present in the 3' region. Rat thymus chromatin does not contain these DNase I hypersensitive sites, suggesting that, in the liver, the chromatin structure is altered so as to allow tissue-specific expression of the P-450c gene. The chromatin structure of the highly inducible P 450c gene is compared to that of the P-450m gene, which is induced to a significantly smaller extent and is constitutively expressed. PMID- 3026445 TI - Dithiothreitol activation of the insulin receptor/kinase does not involve subunit dissociation of the native alpha 2 beta 2 insulin receptor subunit complex. AB - The subunit composition of the dithiothreitol- (DTT) activated insulin receptor/kinase was examined by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis and gel filtration chromatography under denaturing (0.1% SDS) or nondenaturing (0.1% Triton X-100) conditions. Pretreatment of 32P-labeled insulin receptors with 50 mM DTT followed by gel filtration chromatography in 0.1% SDS demonstrated the dissociation of the alpha 2 beta 2 insulin receptor complex (Mr 400,000) into the monomeric 95,000 beta subunit. In contrast, pretreatment of the insulin receptors with 1-50 mM DTT followed by gel filtration chromatography in 0.1% Triton X-100 resulted in no apparent alteration in mobility compared to the untreated insulin receptors. Resolution of this complex by nonreducing SDS polyacrylamide gel electrophoresis and autoradiography demonstrated the existence of the alpha 2 beta 2 heterotetrameric complex with essentially no alpha beta heterodimeric or free monomeric beta subunit species present. This suggests that the insulin receptor can reoxidize into the Mr 400,000 complex after the removal of DTT by gel filtration chromatography. Surprisingly, these apparently reoxidized insulin receptors were also observed to be functional with respect to insulin binding, albeit with a 50% decrease in affinity for insulin and insulin stimulation of the beta subunit autophosphorylation. To prevent reoxidation, the insulin receptors were pretreated with 50 mM DTT followed by incubation with excess N-ethylmaleimide prior to gel filtration chromatography in 0.1% Triton X 100. Under these conditions the insulin receptors migrated as the Mr 400,000 alpha 2 beta 2 complex.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3026446 TI - Electrostatic analysis of the interaction of cytochrome c with native and dimethyl ester heme substituted cytochrome b5. AB - The stability of the complex formed between cytochrome c and dimethyl ester heme substituted cytochrome b5 (DME-cytochrome b5) has been determined under a variety of experimental conditions to evaluate the role of the cytochrome b5 heme propionate groups in the interaction of the two native proteins. Interaction between cytochrome c and the modified cytochrome b5 was found to produce a difference spectrum in the visible range that is very similar to that generated by the interaction of the native proteins and that can be used to monitor complex formation between the two proteins. At pH 8 [25 degrees C (HEPPS), I = 5 mM], DME cytochrome b5 and cytochrome c form a 1:1 complex with an association constant KA of 3 (1) X 10(6) M-1. This pH is the optimal pH for complex formation between these two proteins and is significantly higher than that observed for the interaction between the two native proteins. The stability of the complex formed between DME-cytochrome b5 and cytochrome c is strongly dependent on ionic strength with KA ranging from 2.4 X 10(7) M-1 at I = 1 mM to 8.2 X 10(4) M-1 at I = 13 mM [pH 8.0 (HEPPS), 25 degrees C]. Calculations for the native, trypsin solubilized form of cytochrome b5 and cytochrome c confirm that the intermolecular complex proposed by Salemme [Salemme, F. R. (1976) J. Mol. Biol. 102, 563] describes the protein-protein orientation that is electrostatically favored at neutral pH.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3026447 TI - Kinetics of slow, tight-binding inhibitors of angiotensin converting enzyme. AB - Five phosphorus-containing inhibitors of angiotensin converting enzyme were found to exhibit slow, tight-binding kinetics by using furanacryloyl-L phenylalanylglycylglycine as substrate at pH 7.50 and T = 25 degrees C. Two of the inhibitors, (O-ethylphospho)-Ala-Pro (2) and (O-isopropylphospho)-Ala-Pro (3), are found to follow at minimum a two-step mechanism of binding (mechanism B) to the enzyme. This mechanism consists of an initial fast formation of a weaker enzyme-inhibitor complex (Ki = 130 nM for 2 and 180 nM for 3) followed by a slow reversible isomerization to a tighter complex with measurable forward (K3) and reverse (k4) rate constants (k3 = 4.5 X 10(-2) s-1 for 2 and 5.4 X 10(-2) s-1 for 3; k4 = 9.2 X 10(-3) s-1 for 2 and 3.5 X 10(-3) s-1 for 3). For the remaining three inhibitors, phospho-Ala-Pro (1), (O-benzyl-phospho)-Ala-Pro (4), and (P phenethylphosphono)-Ala-Pro (5), a one-step binding mechanism (mechanism A) is observed under the conditions of the experiment. The second-order rate constants k1 (M-1 s-1) for the binding of these inhibitors to converting enzyme are found to have values more than 3 orders of magnitude lower than the diffusion controlled limit for a bimolecular reaction involving the enzyme, viz., 3.9 X 10(5) for 1, 2.2 X 10(5) for 4, and 4.8 X 10(5) for 5.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3026448 TI - Uncoupling of ATP binding to Na+,K+-ATPase from its stimulation of ouabain binding: studies of the inhibition of Na+,K+-ATPase by a monoclonal antibody. AB - The effects of a monoclonal antibody, prepared against the purified lamb kidney Na+,K+-ATPase, on the enzyme's Na+,K+-dependent ATPase activity were analyzed. This antibody, designated M10-P5-C11, is directed against the catalytic subunit of the "native" holoenzyme. It inhibits greater than 90% of the ATPase activity and acts as a noncompetitive or mixed inhibitor with respect to the ATP, Na+, and K+ dependence of enzyme activity. It inhibits the Na+- and Mg2+ATP-dependent phosphoenzyme intermediate formation. In contrast, it has no effect on K+ dependent p-nitrophenylphosphatase (pNPPase) activity, the interconversion of the phosphoenzyme intermediates, and ADP-sensitive or K+-dependent dephosphorylation. It does not alter ATP binding to the enzyme nor the covalent labeling of the enzyme at the presumed ATP site by fluorescein 5'-isothiocyanate (FITC), but it prevents the ATP-induced stimulation in the rate of cardiac glycoside [3H]ouabain binding to the Na+,K+-ATPase. M10-P5-C11 binding appears to inhibit enzyme function by blocking the transfer of the gamma-phosphoryl of ATP to the phosphorylation site after ATP binding to the enzyme has occurred. In the presence of Mg2+ATP, it also prevents the ATP-induced transmembrane conformational change that enhances cardiac glycoside binding. This uncoupling of ATP binding from its stimulation of ouabain binding and enzyme phosphorylation demonstrates the existence of an enzyme-Mg2+ATP transitional intermediate preceding the formation of the Na+-dependent ADP-sensitive phosphoenzyme intermediate. These results are also consistent with a model of the Na+,K+-ATPase active site being composed of two distinct but interacting regions, the ATP binding site and the phosphorylation site. PMID- 3026449 TI - Isolation of a new vanadium-containing nitrogenase from Azotobacter vinelandii. AB - A new nitrogenase from Azotobacter vinelandii has been isolated and characterized. It consists of two proteins, one of which is almost identical with the Fe protein (component 2) of the conventional enzyme. The second protein (Av1'), however, has now been isolated and shown to differ completely from conventional component 1, i.e., the MoFe protein. This new protein consists of two polypeptides with a total molecular weight of around 200,000. In place of Mo and Fe it contains V and Fe with a V:Fe ratio of 1:13 +/- 3. The ESR spectrum of Av1' also differs from conventional component 1 in that lacks the g = 3.6 resonance that arises from the FeMo cofactor but contains an axial signal with gav less than 2 as well as inflections in the g = 4-6 region possibly arising from an S = 3/2 state. This new enzyme can reduce dinitrogen, protons, and acetylene but is only able to utilize 10-15% of its electrons for the reduction of acetylene. PMID- 3026450 TI - Fe-S centers in lactyl-CoA dehydratase. AB - Lactyl-CoA dehydratase consists of two enzymes, E1 and E2, and requires catalytic quantities of ATP for activity [Kuchta, R. D., & Abeles, R. H. (1985) J. Biol. Chem. 260, 13181-13189]. In contrast to E1, which contains no Fe, E2 contains 8.20 +/- 0.04 mol of Fe/mol of E2, one of which can be removed by 1,10 phenanthroline. E2 also contains 7.33 +/- 0.68 mol of inorganic sulfur/mol of E2, indicating that at least seven of the Fe atoms are present as Fe-S clusters. E1 and E2 contain less than 0.14 mol of Cu, Co, Zn, Mn, and Ni/mol of E1 or E2. Both reduced and oxidized E1 are EPR silent over a 10,000-G scan range at 4 K, while two signals in E2 are observable at 4 K. Identical spectra were obtained with E2 containing either seven or eight Fe atoms, and both signals were only observable at T less than 30 K. Signal 1 has axial symmetry with g = 2.0232 and g = 2.0006. Signal 2 is orthorhombic with g1 = 1.982, g2 = 1.995, and g3 = 2.019. Computer simulation of these spectra with a S = 1/2 spin Hamiltonian was used to extract the g matrices. The intensity of both signals decreases when E2 is reduced with Na2S2O4. We propose that signal 1 is due to an unusual [4Fe-4S] cluster and signal 2 to a [3Fe-3/4S] cluster. Addition of either acrylyl-CoA or lactyl-CoA dramatically alters signal 2.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3026452 TI - Topoisomerase from Ustilago maydis forms a covalent complex with single-stranded DNA through a phosphodiester bond to tyrosine. AB - Highly purified topoisomerase from Ustilago breaks single-stranded DNA, forming a complex with protein covalently bound to the DNA. Methods used to detect the complexes include a nitrocellulose filter assay, electrophoresis of the DNA protein complex in agarose gels containing alkali, and isolation of the complex after removal of all but a small oligonucleotide fragment bound to the protein. The linkage of the Ustilago topoisomerase is to the 3' end of the broken strand of DNA. The DNA-protein complex formed is through a phosphodiester bond to tyrosine. PMID- 3026451 TI - Energetics of cooperative protein-DNA interactions: comparison between quantitative deoxyribonuclease footprint titration and filter binding. AB - Using the binding of cI repressor protein to the lambda right and left operators as a model system, we have analyzed the two common experimental techniques for studying the interactions of genome regulatory proteins with multiple, specific sites on DNA. These are the quantitative DNase footprint titration technique [Brenowitz, M., Senear, D. F., Shea, M. A., & Ackers, G. K. (1986) Methods Enzymol. 130, 132-181] and the nitrocellulose filter binding assay [Riggs, A., Suzuki, H., & Bourgeois, S. (1970) J. Mol. Biol. 48, 67-83]. The footprint titration technique provides binding curves that separately represent the fractional saturation for each site. In principle, such data contain the information necessary to determine the thermodynamic constants for local site binding and cooperativity. We show that in practice, this is not possible for all values of the constants in multisite systems, such as the lambda operators. We show how these constants can nevertheless be uniquely determined by using additional binding data from a small number of mutant operators in which the number of binding sites has been reduced. The filter binding technique does not distinguish binding to the individual sites and yields only macroscopic binding parameters which are composite averages of the various local site and cooperativity constants. Moreover, the resolution of even macroscopic constants from filter binding data for multisite systems requires ad hoc assumptions as to a relationship between the number of ligands bound and the filter retention of the complex. Our results indicate that no such relationship exists. Hence, the technique does not permit determination of thermodynamically valid interaction constants (even macroscopic) in multisite systems. PMID- 3026453 TI - The dnaB protein of Escherichia coli: mechanism of nucleotide binding, hydrolysis, and modulation by dnaC protein. AB - The mechanism of nucleotide binding and hydrolysis by dnaB protein and dnaB X dnaC protein complex has been studied by using fluorescent nucleotide analogues. Binding of trinitrophenyladenosine triphosphate (TNP-ATP) or the corresponding diphosphate (TNP-ADP) results in a blue shift of the emission maximum and a severalfold amplification of the fluorescence emission of the nucleotide analogues. Scatchard analysis of TNP-ATP binding indicates that TNP-ATP binds with a high affinity (Kd = 0.87 microM) and a 8.5-fold enhancement of fluorescence emission of the nucleotide. Only three molecules of TNP-ATP or TNP ADP bind per hexamer of dnaB protein in contrast to six molecules of ATP or ADP binding to a dnaB hexamer. TNP-ATP and TNP-ADP are both competitive inhibitors of single-stranded (SS) DNA-dependent ATPase activity of dnaB protein. TNP-AMP neither binds to dnaB protein nor inhibits the ATPase activity. Formation of dnaB X dnaC complex by dnaC protein results in diminution of the TNP-ATP fluorescence enhancement and a concomitant decrease in the SS DNA-dependent ATPase activity. Kinetic analysis of the ATPase activity of dnaB X dnaC complex indicates that the decrease in the ATPase activity on complex formation is due to a reduction of the maximal velocity (Vmax). The dnaB protein hydrolyzes both TNP-ATP and dATP, however, with an extremely slow rate in the presence of single-stranded M13 DNA. The 2'-OH group of the nucleotide most likely plays an important role in the hydrolysis reaction but not in the nucleotide binding. PMID- 3026454 TI - Nuclease activity of 1,10-phenanthroline-copper ion: reaction with CGCGAATTCGCG and its complexes with netropsin and EcoRI. AB - The self-complementing dodecamer 5'-CGCGAATTCGCG-3' and its complexes with the antibiotic netropsin and the restriction endonuclease EcoRI provide substrates of known three-dimensional structure to study the stereochemistry and mechanism of the artificial nuclease of 1,10-phenanthroline-copper ion [(OP)2Cu+]. Analysis of the reaction products with the 5'-32P dodecamer on 20% sequencing gels has demonstrated the presence of 3'-phosphoglycolate ends in addition to 3' phosphomonoester ends expected from previous studies. A reaction intermediate, which is a precursor to 3'-phosphomonoester termini, has been trapped; in contrast, no comparable species for the 5'-phosphomonoester termini can be detected when 3'-labeled DNAs are utilized as substrates. The reactive oxidative species formed by the coreactants (OP)2Cu+ and hydrogen peroxide is distinguishable in its chemistry from the hydroxyl radicals produced by cobalt-60 gamma-irradiation. The freely diffusible hydroxyl radicals generated by cobalt-60 irradiation produce equivalent amounts of 3'-phosphomonoester and 3' phosphoglycolate termini whereas the 3'-phosphomonoesters are the preferred product of (OP)2Cu+ and H2O2. On the basis of the structures of the products obtained, the principal site of attack of the coordination complex is on the C-1 of the deoxyribose within the minor groove. This conclusion is supported by the footprinting of netropsin binding to the dodecamer. Crystallographic results have demonstrated that netropsin binds to the minor groove at the central AATT residue. A clear protection of attack by the coordination complex at the deoxyriboses associated with A-5, T-6, T-7, and C-9 is fully consistent with attack from the minor groove without intercalation during the course of the cleavage reaction.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3026455 TI - Molecular recognition between oligopeptides and nucleic acids: novel imidazole containing oligopeptides related to netropsin that exhibit altered DNA sequence specificity. AB - Oligopeptides have been synthesized that are structurally related to the antiviral antitumor antibiotic netropsin, but in which each of the pyrrole units is successively replaced by an imidazole moiety, as well as their di- and triimidazole-containing counterparts. These compounds bind to duplex DNA with constants in the range (1.06-1.98) X 10(6) M-1 but not to single-stranded DNA. Since they bind to T4 DNA, it is inferred that, like the parent antibiotic netropsin, they are also minor groove selective. This series of compounds exhibits a progressively decreasing preference for AT sites in binding studies with both native DNAs and synthetic oligonucleotides and a corresponding increasing acceptance of GC base pairs. Footprinting experiments utilizing a 139 base pair HindIII/NciI restriction fragment from pBR 322 DNA revealed that these lexitropsins, or information-reading oligopeptides, recognize more sites than the parent netropsin. In addition, some regions of enhanced nuclease action as the result of drug binding to the fragment were identified. The diimidazole compound in particular recognizes GC-rich sites, implying the formation of new hydrogen bonds between G-C(2)NH2 in the minor groove and the additional N3 imidazole nitrogens. It is clear however that, since the lexitropsins appear to tolerate the original (AT)4 site, an N-methylimidazole group on the ligand will permit either a GC or AT base pair in the binding sequence. Another factor that may be significant in molecular recognition is the high negative electrostatic potential of A X T regions of the minor groove, which is likely to strongly influence binding of these cationic species to DNA.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3026456 TI - Motion and surface accessibility of spin-labeled lipids in a model lipoprotein containing cholesteryl oleate, dimyristoylphosphatidylcholine, and apolipoprotein E. AB - A series of spin-labeled phosphatidylcholines (PCs) and cholesteryl esters (CEs) bearing the paramagnetic 2,2-dimethyloxazolidinyl-1-oxy (doxyl) group at fatty acyl carbon C5', C12', or C16' were used to study acyl chain motions in the polar surface shell and hydrophobic core domains of microemulsion (ME) particles containing cholesteryl oleate and dimyristoylphosphatidylcholine (DMPC), and of particles with apolipoprotein E (apoE) bound to their surfaces. Electron paramagnetic resonance data obtained with the doxyl-labeled PCs indicated a gradient of motion in the ME surface monolayer similar to that observed with the same probes in a bilayer. The 5- and 12-doxyl-CEs clearly demonstrated a higher degree of order for the cholesteryl ester rich core than the corresponding doxyl PCs showed for the phospholipid-rich surface over the entire range 10-60 degrees C. The temperature dependencies of spectra of the 16-doxyl-CE in the core and PC in the surface of the ME were almost identical, suggesting that there was no sharp boundary between core and surface domains. None of the probes detected either the surface phospholipid transition (31 degrees C) or the cholesteryl ester core transition (46 degrees C) measured previously by differential scanning calorimetry and 13C nuclear magnetic resonance. Binding of apoE to spin-labeled DMPC vesicles increased the order of the 5'-position of the sn-2 acyl chain over the range 15-33 degrees C; the thermal transition was broadened and its midpoint elevated. The effect of protein binding was not as striking for the ME particles.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3026457 TI - Kinetic control of mitochondrial ATP synthesis. AB - In order to gain a clearer understanding of the kinetic control of ATP synthesis, rat liver and rat heart mitochondria were incubated under conditions that resulted in various rates of net ATP synthesis or ATP hydrolysis. Radiolabeled phosphate was included in the incubation media, and exchange rates between phosphate and ATP were determined as a function of rates of net ATP synthesis. Since ATP synthase is a highly reversible enzyme, the catalyzed reaction was expected to approach equilibrium especially at low rates of respiration and net ATP synthesis. Thus ADP + Pi V1 in equilibrium V2 ATP. If V1 is the rate of incorporation of radiolabeled phosphate into ATP, then net ATP synthesis (or hydrolysis) is V1 - V2. Since V1 and V1 - V2 could be measured, it was possible to calculate V2. V1 doubled in the transition from zero to maximal net ATP synthesis, whereas V2 decreased by over 90% when the rate of ATP synthesis was high due to high-media ADP. In heart mitochondria at 37 degrees C when respiration increased from 104 +/- 10 to 842 +/- 51 nanoatoms of O2/(min X mg), incorporation of [33P]phosphate into ATP (V1) increased from 1,100 +/- 60 to 1,978 +/- 121 and V2 decreased from 1,100 to near zero. These data demonstrate that mitochondrial ATP synthesis does not occur near equilibrium under physiological conditions and relatively high rates of ATP synthesis. A reaction with a high ratio of forward to reverse flux is obviously not near equilibrium. The important most sensitively controlled reaction appears to be V2, ATP hydrolysis. Possible mechanisms of kinetic control of V2 are discussed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3026459 TI - Production of interleukin 1 by SK hepatoma tumor cells. AB - SK hepatoma cells and SK hepatoma conditioned media contain an 18,000-dalton factor which is pyrogenic, stimulates collagenase and prostaglandin production in skin and synovial fibroblasts, induces bone resorption, and stimulates the proliferation of murine thymocytes. These results are consistent with the finding that this tumor cell line produces interleukin 1 [Doyle, M. V., Brindley, L., Kawasaki, E., & Larrick, J. (1985) Biochem. Biophys. Res. Commun. 130, 768-773] since all these activities have been associated with this cytokine. Greater than 80% of the cellular activity has a molecular weight of 30,000, while in contrast, greater than 80% of the activity in the tumor-conditioned media has a molecular weight of 18,000. When active material from the cells is incubated with trypsin, this high molecular weight material is completely converted into an active 18,000 molecular weight species. The isoelectric point of all active material is always between pI 4.0 and 5.1, regardless of molecular weight. All of these results are consistent with the hypothesis that active, high molecular weight interleukin 1 alpha is first synthesized and stored by the tumor cell. This cytokine is then cleaved by a trypsin-like protease to an active, lower molecular weight species which can be secreted into the media. PMID- 3026458 TI - Spin-label electron paramagnetic resonance and differential scanning calorimetry studies of the interaction between mitochondrial succinate-ubiquinone and ubiquinol-cytochrome c reductases. AB - The interaction between succinate-ubiquinone and ubiquinol-cytochrome c reductases in the purified, dispersed state and in embedded phospholipid vesicles was studied by differential scanning calorimetry and by electron paramagnetic resonance (EPR). When the purified, detergent-dispersed succinate-ubiquinone reductase, ubiquinol-cytochrome c reductase, and cytochrome c oxidase undergo thermodenaturation, they show an endothermic transition. However, when these isolated electron-transfer complexes are embedded in phospholipid vesicles, they undergo exothermodenaturation. The energy released could result from the collapse of the strained interaction between unsaturated fatty acyl groups of phospholipids and an exposed area of the complex formed by removal of interacting proteins. The exothermic enthalpy change of thermodenaturation of a protein phospholipid vesicle containing both succinate-ubiquinone and ubiquinol cytochrome c reductases was smaller than that of a mixture of protein phospholipid vesicles formed from the individual electron-transfer complexes. This suggests specific interaction between succinate-ubiquinone reductase and ubiquinol-cytochrome c reductase in the membrane. This idea is supported by saturation transfer EPR studies showing that the rotational correlation time of spin-labeled ubiquinol-cytochrome c reductase is increased when mixed with succinate-ubiquinone reductase prior to embedding in phospholipid vesicles. These results indicate that succinate-ubiquinone reductase and ubiquinol-cytochrome c reductase are indeed present in the membrane as a supermacromolecular complex. No such supermacromolecular complex is detected between NADH-ubiquinone and ubiquinol-cytochrome c reductases or between succinate-ubiquinone and NADH uniquinone reductases. PMID- 3026461 TI - Purification of a calf thymus DNA-dependent adenosinetriphosphatase that prefers a primer-template junction effector. AB - A purification procedure has been developed that resolves four chromatographically distinct DNA-dependent ATPase activities from calf thymus tissue. One of these activities has been purified to a nearly homogeneous protein, as judged by polyacrylamide gel electrophoresis. This protein has a specific activity of 18 mumol of ATP hydrolyzed per minute per milligram of protein and is active only in the presence of a DNA effector. The DNA-dependent ATPase activity is greatest in the presence of DNA containing a 3'-hydroxyl primer-template junction with a segment of adjacent single strand, i.e., a DNA polymerase substrate. Primer-template effectors that have had the 3'-hydroxyl group eliminated by the addition of a dideoxyribonucleotide are less active as cofactors for ATP hydrolysis than effectors which retain the 3'-hydroxyl group. Other DNAs can serve as cofactors, but with a reduced rate of ATP hydrolysis. DNA cofactors which are single stranded are much more effective at promoting ATPase activity than completely double-stranded cofactors, although the effectiveness of single-stranded DNA decreases as the length of the oligonucleotide decreases. An RNA/DNA hybrid does not promote ATPase activity. These data suggest that ATPase A may be involved in the recognition of primer-template junctions and the elongation phase of DNA synthesis. PMID- 3026460 TI - Effects of hydration on purine motion in solid DNA. AB - Deuterium quadrupole echo spectra and spin-lattice relaxation rates measured at 76.8 and 38.4 MHz as a function of relative humidity are reported for calf thymus DNA deuterated at positions A8 and G8. The amplitude of base pair motion is observed to increase slightly with increasing degree of hydration (up to approximately 20 mol of H2O/nucleotide), and the onset of motion is associated with a more than 100-fold drop in T1. This observed decrease in T1 parallels that observed previously for the phosphate backbone and appears to be characteristic of collective modes of motion. Above approximately 20 mol of H2O/nucleotide, the amplitude of the base motion increases substantially up to a point where slow components of motion lead to a complete loss of the quadrupole echo. PMID- 3026462 TI - DNA binding specificity of a series of cationic metalloporphyrin complexes. AB - The sequence specificities of a series of cationic metalloporphyrins toward a 139 base pair restriction fragment of pBR-322 DNA have been studied by DNase I footprinting methodology. Analysis using controlled digests and quantitative autoradiography/microdensitometry revealed that the 5- and 6-coordinate complexes of meso-tetrakis(N-methyl-4-pyridiniumyl)porphine, MT4MPyP, where M is Mn, Fe, Co, and Zn, were found to bind to AT regions of DNA. Footprinting analysis involving the radiolabel on the opposing strand of restriction fragment showed site skewing in the direction of the 3' end of the fragment, indicating that the porphyrins bind in the minor groove of DNA. The significant increase in DNase I catalyzed hydrolysis observed in various regions of the fragment appeared to be primarily due to a decrease in available substrate DNA upon porphyrin binding with possible contributions from structural changes in DNA caused by ligand binding. The complexes NiT4MPyP and CuT4MPyP were found to bind to both AT and GC regions of the fragment, producing different degrees of inhibition in the two regions. Since the outside-binding porphyrins can neither intercalate or effectively hydrogen bond to DNA, they appear to read sequence by responding to steric and/or electrostatic potential effects located in the minor groove of DNA. PMID- 3026463 TI - Cloning and characterization of the gene for rabbit C-reactive protein. AB - C-reactive protein (CRP), an acute-phase plasma protein of hepatic origin in man and rabbit, is a cyclic pentamer composed of five identical nonglycosylated Mr 22 500 subunits. We have isolated both cDNA and genomic clones for rabbit CRP. These clones were used as probes to demonstrate that when CRP synthesis is increased following an acute inflammatory stimulus, there is a corresponding increase in the level of accumulated CRP mRNA. The rabbit CRP gene is 2.6 kilobases in length containing a single intron of 252 base pairs (bp) which interrupts the codon for amino acid 2 in the protein. The mRNA for CRP contains a 5'-nontranslated region of 113 bp and a 3'-nontranslated region of 1550 bp. Sequencing of the protein coding region of the gene indicates that the primary translation product contains a 20 amino acid N-terminal signal peptide. The deduced amino acid sequence is in general agreement with the published sequence [Wang, C. M., Nguyen, N. Y., Yonaha, K., Robey, F., & Liu, T.-Y. (1982) J. Biol. Chem. 257, 13610-13615] except in the region between amino acids 63 and 73. In this region, the sequence of both cDNA and genomic clones indicates the presence of 28 amino acids not previously reported. This alteration may be the result of genetic heterogeneity or an error in the reported protein sequence. PMID- 3026464 TI - Fusion of phospholipid vesicles induced by alpha-lactalbumin at acidic pH. AB - Alpha-Lactalbumin (alpha-LA), lysozyme, and ribonuclease are found to induce fusion of phosphatidylserine/phosphatidylethanolamine vesicles at low pH. The fusogenic behavior and the binding to phospholipid vesicles of one of these proteins, alpha-LA, are studied at a wide range of conditions. The initial rate of fusion in the presence of alpha-LA increases with increasing acidity below pH 6, and the extent of alpha-LA binding to the vesicles is also found to increase with decreasing pH. Once bound to the vesicles in acidic media, the neutralization to pH 7 fails to dislodge the alpha-LA from the vesicles, and this irreversible binding also increases with decreasing pH. A segment of alpha-LA is found to be resistant to the proteolytic digestion when initially incubated with the vesicles at low pH. The amino acid composition of this fragment was determined, and from this the sequence of alpha-LA fragment, which appears to be inserted into the bilayer, is deduced. Hydrophobic labeling with dansyl chloride renders support that this segment indeed penetrates into the hydrophobic interior of bilayer. Since both the N-terminal and the C-terminal of this vesicle-bound protein are accessible to the externally added proteolytic enzymes, it is concluded that a loop of the polypeptide segment goes into the bilayer. These observations, taken together, suggest a possibility that the penetration by a loop of alpha-LA segment into the phospholipid bilayer is responsible for the fusion. PMID- 3026466 TI - Inhibition of beta nerve growth factor binding to PC12 cells by alpha nerve growth factor and gamma nerve growth factor. AB - Pheochromocytoma (PC12) cells have been found to differ from dorsal root ganglionic cells with respect to the modulation of the beta nerve growth factor (beta NGF) binding properties elicited by alpha NGF and gamma NGF. In contrast to our previous results with intact dorsal root ganglionic cells in which only high affinity binding was blocked, alpha NGF and gamma NGF were found to block competitively all steady-state binding of iodinated beta NGF to PC12 cells at both 37 and 0.5 degrees C. The EC50 that was found for the alpha NGF displacement was 9-10 microM, and the gamma NGF effect had an EC50 of 200 nM, in the predicted range based upon the apparent Kd for dissociation of the alpha beta or the beta gamma complex in solution. The concurrence of the binding EC50 and the Kd for each complex indicates that the formation of alpha beta or beta gamma complexes in solution competes with the process of PC12 receptor binding with 125I-beta NGF. Experiments were carried out examining the dissociation kinetics following the addition of excess unlabeled beta NGF or alpha NGF at both 37 and 0.5 degrees C. Three dissociation components were observed with alpha NGF, in contrast to the two normally found with beta NGF. Lowering the chase temperature to 0.5 degrees C changed the relative contributions made by each component without dramatically changing any of the rate constants. The "slow" receptor was further examined by the dependence on 125I-beta NGF concentration of the slowest component with a chase of either excess alpha NGF or excess gamma NGF at 0.5 degrees C.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3026465 TI - Beta nerve growth factor binding to PC12 cells. Association kinetics and cooperative interactions. AB - The association kinetics of 125I beta nerve growth factor (NGF) binding to the PC12 clonal cell line have been examined in detail at 0.5 and 37 degrees C. These data were examined by utilizing a reversible second-order integrated rate equation, and the results were not consistent with a simple bimolecular process. Two association rates were required to explain the results adequately. At 37 degrees C, the faster component was estimated to have a second-order association rate constant of 1.4 X 10(7) M-1 s-1, while the rate constant for the slower component (3.8 X 10(6) M-1 s-1) was about 4-fold lower. As shown by others, the temperature dependence of the dissociation kinetics indicated that while the rapidly dissociating component was only slightly slowed by lowering the chase temperature to 0.5 degrees C, the second component was slowed by about 270-fold, from 8 X 10(-4) s-1 to 3 X 10(-6) s-1. The binding data that describe the slowly dissociating component were obtained by utilizing this differential temperature dependence and revealed a concave downward Scatchard plot. The binding parameters determined from computer analysis using a nonlinear fitting program (LIGAND) suggest that this component consists of (a) an interacting class of about 4000 sites/cell that have a first stoichiometric steady-state dissociation constant of 65 pM and a second stoichiometric interaction constant of 16 pM, indicative of positively cooperative interactions, and (b) a class of sites consistent with a ratio of sites/Kd of about 11.1 sites/(cell X pM). The steady-state binding results at 37 degrees C indicated only one class of binding sites (155,000 +/- 18,000 sites/cell) that had an apparent Kd of 0.52 +/- 0.03 nM. One class of sites was also observed at 0.5 degrees C, and the receptor concentration was found to be reduced (99,000 +/- 7600 sites/cell) while the Kd was increased (1.7 +/- 0.14 nM). A significant level of positively cooperative interactions was observed frequently at 37 degrees C that was not due to a failure to reach steady state conditions, internalization, or degradation. Since cooperativity of binding was never observed at 0.5 degrees C, a membrane event may be involved. Determination of the contribution of the different classes of NGF receptors found on PC12 cells to the biological actions of NGF requires a clear understanding of their kinetic properties and their relationship to each other. The studies presented here indicate that their interactions are more complex than previously described. PMID- 3026467 TI - Role of free radical processes in stimulated human polymorphonuclear leukocytes. AB - Human polymorphonuclear leukocytes produce large quantities of superoxide when they attack and kill bacteria. However, superoxide is a weak oxidizing and reducing agent, and other more reactive oxygen species derived from reactions of superoxide are suggested to participate in the killing processes. To test the hypothesis that a reactive free radical or singlet oxygen is involved in bactericidal activity, human polymorphonuclear leukocytes were exposed to phagocytozable particles containing lipids that contain the easily autoxidized 1,4-diene moiety. After incubation the preparations were extracted and the extracts reduced with NaBH4 to convert hydroperoxides to stable alcohols. Using gas chromatography/mass spectrometry to analyze the extracts, we were unable to detect products unless iron salts were added to the medium. The products obtained by extraction are those that would be expected if both free radical chain autoxidation and 1O2 oxidation were taking place. In summary, we find that polymorphonuclear leukocytes do not cause peroxidation, implying that formation of strongly oxidizing free radicals is not an intrinsic property of the leukocyte. Added iron catalyzes peroxidation by activated leukocytes yielding an unusual distribution of hydroxylated products. PMID- 3026468 TI - A new acylphosphatase isoenzyme from human erythrocytes: purification, characterization, and primary structure. AB - A new acylphosphatase from human erythrocytes was isolated by an original purification procedure. It is an isoenzyme of the well-characterized human skeletal muscle acylphosphatase. The erythrocyte enzyme shows hydrolytic activity on acyl phosphates with higher affinity than the muscle enzyme for some substrates and phosphorylated inhibitors. The sequence was determined by characterizing the peptides purified from tryptic, peptic, and Staphylococcus aureus V8 protease digests of the protein, and it was found to differ in 44% of the total positions as compared to the human muscle enzyme. About one-third of these differences are in the form of strictly conservative replacements. The protein consists of 98 amino acid residues; it has an acetylated NH2-terminus and does not contain cysteine: (sequence in text). PMID- 3026469 TI - The role of fixed and mobile buffers in the kinetics of proton movement. AB - We derive a simple expression for the effective diffusion coefficient of protons in Fick's second law, Deff, when both spatially fixed, HF, and mobile, HM, buffers are present. These buffers are present at moderately high concentrations ([Ftot], [Mtot] greater than 1 mM) in most biological systems. We consider only the case where the protonation reactions remain at equilibrium during the diffusion process. When the pH is to the alkaline side of the pK values of the fixed and mobile buffers ([H+] less than KF, KM), the effective diffusion coefficient of protons in Ficks second law is: (Formula: see text) where DH is the diffusion coefficient of the protons free in the aqueous phase and DHM is the diffusion coefficient of the mobile buffer. The equation illustrates three features of diffusion in a buffered system. Firstly, the effective diffusion coefficient of protons is always lower than the diffusion coefficient of free protons. Secondly, increasing the concentration of fixed buffers always decreases Deff. Thirdly, increasing the concentration of mobile buffer can increase Deff when fixed buffers are present. PMID- 3026470 TI - Human cytochrome c oxidase isoenzymes from heart and skeletal muscle; purification and properties. AB - Human cytochrome c oxidase was isolated in an active form from heart and from skeletal muscle by a fast, small-scale isolation method. The procedure involves differential solubilisation of the oxidase from mitochondrial fragments by laurylmaltoside and KCl, followed by size-exclusion high-performance liquid chromatography. Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate showed differences between the subunit VI region of cytochrome c oxidases from human heart and skeletal muscle, suggesting different isoenzyme forms in the two organs. This finding might be of importance in explaining mitochondrial myopathy which shows a deficiency of cytochrome c oxidase in skeletal muscle only. In SDS polyacrylamide gel electrophoresis most human cytochrome c oxidase subunits migrated differently from their bovine counterparts. However, the position of subunits III and IV was the same in the human and in the bovine enzymes. The much higher mobility of human cytochrome c oxidase subunit II is explained by a greater hydrophobicity of this polypeptide than of that of the subunit II of the bovine enzyme. PMID- 3026473 TI - Furosemide-sensitive Na+ and K+ transport and human erythrocyte volume. AB - The relationship between cation transport and cell volume in human erythrocytes was investigated by measuring ouabain-sensitive K+ influx, ouabain-resistant, furosemide-sensitive K+ influx, and ouabain + furosemide-resistant K+ influx, and maximal ouabain binding in microcytic, normocytic and macrocytic red cells. A significant correlation was found between the mean corpuscular volume and furosemide-sensitive K+ influx normalized either to cell number (r = 0.636, P less than 0.001) or to cell volume (r = 0.488, P less than 0.001). No relationship was seen between mean corpuscular volume and ouabain-sensitive K+ influx, and the number of ouabain-binding sites per cell was only weakly correlated with mean corpuscular volume (r = 0.337, P less than 0.05). A slight, negative relationship existed between mean corpuscular volume and ouabain + furosemide-resistant K+ influx expressed per volume of cells (r = -0.359, P less than 0.01), and an apparent relationship between furosemide-sensitive K+ influx and mean corpuscular hemoglobin concentration (r = 0.446, P less than 0.01) disappeared when microcytic samples were excluded from analysis. Furosemide sensitive transport, including Na+ influx and K+ and Na+ efflux, was completely absent in microcytic cells from one patient with alpha-thalassemia minor. In addition, these cells exhibited a furosemide-resistant, Cl(-)-dependent K+ influx. Exposure of normal erythrocytes to hypotonic conditions (196 mosM) increased furosemide-sensitive K+ influx by a mean of 45% (P less than 0.05), while exposure to hypertonic conditions (386 mosM) had no significant effect. The results indicate that furosemide-sensitive transport and cell volume are interrelated in human erythrocytes. However, the inability to fully recreate this relationship with in vitro manipulation of cell volume suggest that this relationship is established prior to red cell maturation. PMID- 3026471 TI - Phenylglyoxal modification of the Photosystem II reaction center. AB - Time-resolved spectroscopic techniques, including optical flash photolysis and electron spin resonance (ESR), have been used in conjunction with fluorescence induction and dye-reduction assays to monitor electron transport in Photosystem II (PS II) subchloroplast particles incubated with the covalent modifier, phenylglyoxal. Phenylglyoxal-modified digitonin (D-10) particles from spinach are characterized by a high initial fluorescence yield (Fi) and an abolition of the variable component of fluorescence (Fv); an inhibition of PS-II-mediated reduction of dichlorophenol indophenol (DPIP) by sym-diphenylcarbazide; an abolition of flash-induced absorption transients (t1/2 greater than 2 microseconds) at 820 nm attributed to the primary electron donor, P-680+; the inhibition of photoreduction of the acceptor Qa; and the elimination of the ESR Signal 2s and Signal 2f. These observations suggest the critical participation of specific arginine residues on both the oxidizing and reducing sides of Photosystem II and also implicate phenylglyoxal as a quinone-binding site inhibitor (Golbeck, J.H. and Warden, J.T. (1984) Biochim. Biophys. Acta 767, 263 271). PMID- 3026472 TI - Modification of membrane physical properties, biological response and insulin binding in Friend cells by low serum concentration. AB - The effect of low serum concentration on plasma membrane fluidity and lipid composition, differentiation and insulin binding was investigated in three Friend erythroleukemia clones. Both FLC (clones No. 745) and F(+) (Ostertag F4N) Friend erythroleukemia cells can be induced to differentiate and to produce hemoglobin when exposed to DMSO. Clone R(3) (Ostertag F4-D5-1) is a DMSO-resistant clone when grown under normal conditions (15% serum) but could undergo differentiation with accumulation of protoporphyrin IX upon induction with DMSO when grown in low serum concentration (2.5% serum). Electron spin resonance measurements of the order parameters (S) and S(T parallel) demonstrate that R(3) has a more fluid plasma membrane than the FLC and F(+). The order parameters of the outer hyperfine splittings S(T parallel) at 37 degrees C are 0.60 +/- 0.009, 0.62 +/- 0.008 and 0.64 +/- 0.009 for R(3), F(+) and FLC, respectively. We have used the insulin receptors as a model for a polypeptide hormone receptor associated with the plasma membrane of the Friend clones. Insulin binding studies demonstrated that the receptor of R(3) had a decreased affinity for insulin manifest as a 10 fold increase in the amount of insulin required to compete for half of the tracer binding (18 nM for R(3) vs. 2 nM for FLC and F(+)). Computer-fit Scatchard plot analysis by the negative cooperativity model reveal that R(3) possessed a similar number of sites/cell (about 70,000) as the FLC or F(+) cells, with similar high and low affinities. Growing the DMSO-resistant clone R(3) in low serum concentration caused a decrease in receptor number by 35%, and an increase in receptor affinity to that seen with the differentiable clones. Thus, the abnormal properties of the plasma membrane and insulin receptor of the DMSO-resistant clone in our earlier report (Simon et al. (1984) Biochim. Biophys. Acta 803, 39 47) were partially reversed by growing the cells in a low serum concentration, restoring the cellular response to the differentiation agent. PMID- 3026474 TI - Neurotensin receptors in canine intestinal smooth muscle: preparation of plasma membranes and characterization of (Tyr3-125I)-labelled neurotensin binding. AB - To study the binding of (Tyr3-125I)-labelled neurotensin to intestinal muscle, plasma membranes have been purified from dog intestinal circular smooth muscle. Purification was done by differential centrifugation followed by separation on a sucrose gradient. Electron microscopic study revealed that the dissected circular muscles used as the source of membranes were free of myenteric plexus and that the plasma membrane fraction obtained was free of any mitochondria or synaptosomes. The fraction used was obtained at the interface of 14%-33% sucrose density on the gradient and was 25-times enriched in the plasma membrane marker enzyme 5'-nucleotidase activity as compared to post-nuclear supernatant. This fraction contained negligible activity of mitochondrial membrane marker enzyme cytochrome c oxidase and low activity of a putative endoplasmic reticulum marker enzyme NADPH-cytochrome-c reductase. This membrane fraction contained a high density of neurotensin binding sites. This binding was studied by kinetic and by saturation approaches. Analysis of data from saturation binding studies by the computer programs (EBDA and LIGAND) suggested the presence of a two-site model (Kd1 = 0.118 nM, Kd2 = 3.18 nM, Bmax1 = 9.73 fmol/mg and Bmax2 = 129.8 fmol/mg). A part of specifically bound neurotensin was rapidly dissociated. No cooperativity between the two receptor types could be detected. A kinetic analysis of binding gave the Kd value equal to 0.107 nM. Carboxy terminal amino acid residues 8-13 were found to be essential for the binding activity and replacement of Tyr11 by tryptophan reduced the affinity of the peptide by 10 times in displacement studies. Binding was modulated by sodium ions and a guanine nucleotide Gpp[NH]p. MgCl2, CaCl2 and KCl were also found to reduce the specific binding. Evidence was found of a high specific binding to another membrane fraction poor in plasma membranes and rich in synaptosomes. We concluded that plasma membrane of canine intestinal circular muscle contains neurotensin receptors with recognition properties distinct from those obtained in previous studies of neurotensin binding sites in murine tissues. Another neurotensin binding site may be present on neuronal membranes. PMID- 3026475 TI - Preferential lipid association and mode of penetration of apocytochrome c in mixed model membranes as monitored by tryptophanyl fluorescence quenching using brominated phospholipids. AB - The fluorescence of the single tryptophan residue at position 59 in apocytochrome c, the biosynthetic precursor of the inner mitochondrial membrane protein cytochrome c, was studied in small unilamellar vesicles composed of phosphatidylserine (PS) and phosphatidylcholine (PC) with or without specifically Br-labelled acyl chains at the sn-2 position. The protein has a very high affinity for PS-containing vesicles (dissociation constant Kd less than 1 microM). From the relative quenching efficiency by the brominated phospholipids, it could be concluded that the protein specifically associates with the PS component in mixed vesicles and that maximal quenching occurred with phospholipids in which the bromine was present at the 6,7-position of the 2-acyl chain suggesting that (part of) the bound protein penetrates 7-8 A deep into the hydrophobic core of the bilayer. PMID- 3026477 TI - Inhibition of (H+ + K+)-ATPase by omeprazole in isolated gastric vesicles requires proton transport. AB - Omeprazole was found to inhibit the (H+ + K+)-ATPase activity in isolated gastric vesicles only when acid was accumulated in the vesicle lumen. The ATPase activity was time- and dose-dependently inhibited in the presence of K+ and valinomycin. Under conditions in which no pH-gradient was generated, i.e., in the presence of K+ alone or NH4+, no effect of omeprazole was found. The degree of inhibition was directly correlated to the amount of inhibitor bound to the preparation. A stoichiometry of 2 mol radiolabelled inhibitor bound per mol phosphoenzyme was found on total inhibition of the K+ plus valinomycin-stimulated activity. This inhibitory action of omeprazole on the ATPase activity could be fully reversed by addition of beta-mercaptoethanol. The inhibition of the proton transport in the (H+ + K+)-ATPase-containing vesicles by omeprazole was also strictly correlated to the amount of bound inhibitor. The stoichiometry of binding at total inhibition of this reaction was found to be 1.4 mol per mol phosphoenzyme. The K+ stimulated p-nitrophenylphosphatase activity was inhibited in parallel with the ATPase activity, whereas the phosphoenzyme levels were affected to a lesser extent by omeprazole. Gel electrophoresis of an omeprazole-inhibited vesicle preparation showed that the radiolabel was mainly found at 94 kDa, the molecular weight of the (H+ + K+)-ATPase catalytic subunit(s). PMID- 3026476 TI - The role of protons in the mechanism of galactoside transport via the lactose permease of Escherichia coli. AB - The kinetic mechanism of lactose transport across the cytoplasmic membrane has been investigated and the results related to standard models for the lactose-H+ symport reaction using computer simulation. It is shown that the biphasic kinetics reported for lactose uptake (Kaczorowski, G.J. and Kaback, H.R. (1979) Biochemistry 18, 3691-3697) are consistent with random binding of lactose and protons and rapid subsequent translocation of the ternary lactose-H+-permease complex. Such a model is also shown to explain the observed dependence of the kinetic parameters on the magnitude of the protonmotive force. Both sugar and protons are shown to cause product inhibition of lactose flux and the ability of standard models to account for the pattern of inhibition is discussed. Three apparent dissociation constants have been determined for the protonation reactions in the external medium: two (pKa 6.3 and 9.6) control the activity of the permease, whilst the third (pKa 8.3) controls the affinity of the permease for galactosides. A similar set of dissociation constants has been determined for the internal reactions. Again two (pKa 6 and 9.8) control activity and a third (pKa 8.8) controls the affinity for galactosides. The dissociation reactions characterised by pKa 8.3, 8.8, 9.6 and 9.8 are attributed to the dissociation of the substrate (symported) proton from the binary proton-permease complexes (pKa 8.3 and 8.8) and the ternary proton-galactoside-permease complexes (pKa 9.6 and 9.8). The third pair (pKa 6.3 and 6.0) must be interpreted as describing a separate protonation reaction which may have a regulatory or auxiliary role in transport. PMID- 3026478 TI - Kinetic and topographical studies of the phosphatidylcholine: ceramide choline phosphotransferase in plasma membrane particles from mouse ascites cells. AB - The content of endogenous phospholipids in plasma membrane preparations from Ehrlich ascites cells was depleted by exposure to phospholipase C. The enzyme catalyzing the phosphatidylcholine: ceramide choline phosphotransferase reaction was inactivated by this treatment. However, the activity could be restored with exogenous phosphatidylcholines, demonstrating the dependence of the reaction upon the presence of this substrate. Phosphatidylcholines containing unsaturated fatty acids were 10-fold more effective substrates than the saturated molecular species. The activation energy of the reaction was determined to be 17.2 kcal/mol. Selective trypsin treatment of the plasma membranes suggests that the cholinephosphotransferase may have an asymmetric orientation. The reaction kinetics followed a rate equation similar to that of the ping-pong reaction mechanism, which suggests the formation of an enzyme-bound intermediate of the phosphocholine group being transferred. These results are discussed in terms of possible biological functions of the enzyme. PMID- 3026479 TI - A model for accelerated uptake and accumulation of sugars arising from phosphorylation at the inner surface of the cell membrane. AB - A model transport system for cellular accumulation of sugar coupled to phosphorylation is described. Sugar permeates the cell membrane via a passive facilitated transport system. On the inside surface of the membrane the bound sugar is either phosphorylated to form impermeable hexose phosphate, which is released into the intracellular solution, or released directly into the cytosol. Sugar may be regenerated from hexose phosphate in the cytosol via a phosphatase reaction. The reduction of the proportion of sites on the inner membrane surface occupied by permeable sugar, caused by the kinase reaction, increases both net and unidirectional passive inflow and reduces both net and unidirectional exit of sugar, thereby permitting large stationary state gradients of free sugar to be maintained between the cytosol and bathing solution. In cells where there is a high passive membrane permeability to free sugar, steady-state accumulation of free sugar within the cytosol, linked to metabolism is inexplicable in terms of conventional transport kinetics based on equilibrium thermodynamic assumptions. This phenomenon is analysed in terms of non-equilibrium stationary state flows of ligands via a probability network. The effects of metabolism on exchange transport are also examined. The model provides a framework to explain how sugar transport is loosely coupled to phosphorylation in mammalian epithelial cells, adipocytes, yeasts and bacteria, so that a high rate of substrate accumulation is maintained without requiring a reduction in the intracellular concentration of permeable substrate below that in the external solution. PMID- 3026480 TI - Genetic alterations of integrated avian sarcoma virus DNA sequences in transformed rat cells. AB - Clones and subclones of Schmidt-Ruppin-RSV-D-(SRD-) infected rat cells that were isolated in soft agar have shown differences in their morphology. Some of the subclones were round or spindle-shaped cells with lower anchorage dependence and high growth rate, while others were more fibroblast-like and failed to overgrow one another. In correlation to their morphology, subclones with high degree of phenotypic transformation contained amplified amounts of full length proviral DNA or proviral fragments. The amplification affected the proviral DNA elements together with host cellular DNA sequences. We have also shown a relationship between the copy number of the proviruses in the cells, the level of expression of viral RNA, and between those two parameters and the various degrees of phenotypic transformation. PMID- 3026481 TI - The glyceraldehyde-3-phosphate dehydrogenase gene family in the nematode, Caenorhabditis elegans: isolation and characterization of one of the genes. AB - The isolation and genomic sequence of one of possibly four glyceraldehyde-3 phosphate dehydrogenase genes in the nematode, Caenorhabditis elegans is presented. The complete nucleotide sequence of the coding as well as the noncoding flanking regions of this gene has been determined. The deduced amino acid sequence agrees with the sequence of typical glyceraldehyde-3-phosphate dehydrogenase enzymes and its molecular weight of 36,235 agrees with its size determined previously (Yarbrough, P. and Hecht, R. (1984) J. Biol. Chem. 259, 14711-14720). That this isolated gene encodes a nematode glyceraldehyde-3 phosphate dehydrogenase is additionally confirmed by demonstrating its immunoreactivity to an anti-nematode glyceraldehyde-3-phosphate dehydrogenase antibody after its expression as a fusion protein with dihydrofolate reductase. Codon utilization follows a pattern typical of other expressed nematode genes. The gene is split by two introns that are highly conserved in comparison to other introns observed in C. elegans. The placement of one of these introns is conserved with respect to the chicken glyceraldehyde-3-phosphate dehydrogenase gene. Within the 5' flanking sequence homology to actin and the homology 2 block of the major myosin gene (unc-54) is noted. It is of interest that the 3' flanking region contains a CAAAT box, followed by a TATAAT box, before an open reading frame of a closely linked gene that also contains a small AT-rich intron with the nematode consensus splice junction. PMID- 3026482 TI - Site-specific gap-misrepair mutagenesis by O4-ethylthymine. AB - The mutagenicity of the DNA base O-alkylation adduct, O4-ethylthymine, specifically incorporated into the plasmid vector pUC8 at the unique SalI and HincII recognition sites, was studied in vivo. Escherichia coli, Micrococcus luteus and AMV DNA polymerases catalyze the incorporation of O4-ethylTMP against template adenine and guanine residues, resulting in DNA sequence alteration during subsequent replication in the host E. coli K-12 strain JM83. The greatest mutation frequency was observed with error-prone AMV DNA polymerase. High levels of cognate restriction endonuclease-resistant mutant plasmid isolates were obtained by gap replication repair in the presence of O4-ethylTTP. The yields of mutant isolates were dependent upon the relative concentration of the competing pyrimidine deoxynucleoside triphosphates, TTP and dCTP, in the misreplication reaction. Repair of incorporated O4-ethylTMP of plasmid DNA by in vitro treatment with specific alkyltransferase, prior to transformation in the host, effectively increases the mutagenic efficiency of the adduct. The results obtained are consistent with the high miscoding potential O4-ethylthymine observed in in vitro studies and its ability to base-pair with noncomplementary guanine residues in DNA. PMID- 3026483 TI - The cleavage of single-stranded DNA by the isoschizomeric restriction endonuclease HhaI and CfoI. AB - The cleavage of single-stranded (ss) M13mp8(+) DNA by the isoschizomeric restriction endonucleases HhaI and CfoI has been investigated. The two enzymes differ considerably in their ability to cleave ssDNA. HhaI cleaves ssDNA about two orders of magnitude faster than does CfoI, although both enzymes show the same activity when assayed on double-stranded DNA. From the cleavage of oligonucleotides and of M13mp8(+) DNA fragments it is concluded that cleavage of ssDNA occurs via transiently formed double-stranded hairpin structures. A rough correlation exists between the stability of the secondary structures and the cleavage efficiency. PMID- 3026484 TI - Stereochemical considerations in the enzymatic phosphorylation and antiviral activity of acyclonucleosides. I. Phosphorylation of 2'-nor-2'-deoxyguanosine. AB - The antiviral compound 9-[(1,3-dihydroxy-2-propoxy)methyl]guanine (2'-nor-2' deoxyguanosine, 2'-NDG) is phosphorylated by the HSV-1-induced thymidine kinase to the monophosphate (2'-NDG-MP) and this is further phosphorylated by cellular kinases to the triphosphate (2'-NDG-TP) which is a potent inhibitor of DNA polymerases. Since phosphorylation of 2'-NDG creates a chiral center in the molecule, it was of interest to examine whether both monophosphate enantiomers were produced by the viral thymidine kinase, whether they both could be further phosphorylated by cellular kinases and, if so, whether the respective triphosphates were equally inhibitory to the DNA polymerases. The time course of the phosphorylation by GMP kinase of a chemically synthesized, racemic 2'-NDG-MP was compared to that of a 2'-NDG-MP preparation obtained by enzymatic phosphorylation of 2'-NDG with HSV-1 thymidine kinase. The results indicated that the two enantiomeric monophosphates were phosphorylated by GMP kinase with different rates and that phosphorylation of 2'-NDG by HSV-1 thymidine kinase gave only one of the isomers, whose structure was determined to be S. Both enantiomeric diphosphates were further phosphorylated to the respective triphosphates and it was shown that, in contrast to the triphosphate obtained from the 2'-NDG-MP prepared by viral thymidine kinase which was a potent inhibitor of HSV-1 DNA polymerase, the triphosphate obtained from the slow reacting R isomer had little or no inhibitory activity against this enzyme. PMID- 3026485 TI - Circular dichroism studies on a synthetic peptide corresponding to the membrane spanning region of vesicular stomatitis virus G protein and its fatty acyl derivative. AB - The conformations of synthetic peptides Lys-Phe-Phe-Phe-Ile-Ile-Gly-Leu-Ile-Ile Gly-Leu-Phe-OCH3 and Lys(epsilon-palmitoyl)-Phe-Phe-Phe-Ile-Ile-Gly-Leu-Ile-Ile Gly-Leu-Phe-O CH3, which constitute a part of the membrane-spanning region of the vesicular stomatitis virus G protein, have been studied by circular dichroism (CD) spectroscopy. Secondary structural features are observed for both peptides in trifluoroethanol, methanol, aqueous mixtures of trifluoroethanol and methanol and in a micellar environment. In trifluoroethanol, the CD spectra indicate the presence of a helical conformation, whereas in aqueous mixtures of organic solvents, both helical and beta-conformations are observed. While fatty acid acylation does not directly modulate peptide conformation, it promotes self association of the acylated peptide and association with micelles. In a micellar environment, the acylated peptide adopts an alpha-helical conformation. PMID- 3026487 TI - Early effect of myo-inositol deficiency on phosphatidylinositol metabolism in rat liver. AB - Young rats (100 g) were fed either a myo-inositol-deficient or supplemented (control) diet for up to 14 days following a 12 h fast. At various times during this period animals were killed, livers were removed, and a microsomal fraction was prepared and assayed for CDPdiacylglycerol inositol transferase activity and for phosphatidylinositol-inositol exchange activity. Within 2 days after beginning the regimen, rats consuming the deficient diet had a 40% lower activity of the transferase than rats consuming the control diet. This difference was maintained throughout the feeding period and developed simultaneously with the accumulation of triacylglycerol in the deficient livers. In contrast, the specific activity of the exchange enzyme was unchanged by feeding the deficient diet. PMID- 3026486 TI - Characterization of the specific binding of rat apolipoprotein E-deficient HDL to rat hepatic plasma membranes. AB - We have used a preparation of rat liver plasma membranes to study the binding of rat apolipoprotein E-deficient HDL to rat liver. The membranes were found to bind HDL by a saturable process that was competed for by excess unlabeled HDL. The binding was temperature-dependent and was 85% receptor-mediated when incubated at 4, 22 and 37 degrees C. The affinity of the binding site for the HDL was consistent at all temperatures, while the maximum binding capacity increased at higher temperatures. The specific binding of HDL to the membranes did not require calcium and was independent of the concentration of NaCl in the media. The effect of varying the pH of the media on HDL binding was small, being 30% higher at pH 6.5 than at pH 9.0. Both rat HDL and human HDL3 were found to compete for the binding of rat HDL to the membranes, whereas rat VLDL remnants and human LDL did not compete. At 4 degrees C, complexes of dimyristoylphosphatidylcholine (DMPC) and apolipoproteins A-I, A-IV and the C apolipoproteins, but not apolipoprotein E, competed for HDL binding to the membranes. At 22 and 37 degrees C, all DMPC apolipoprotein complexes competed to a similar extent, DMPC vesicles that contained no protein did not compete for the binding of HDL. These results suggest that the rat liver possesses a specific receptor for apolipoprotein E deficient HDL that recognizes apolipoproteins A-I, A-IV and the C apolipoproteins as ligands. PMID- 3026488 TI - Effect of spermine on the translocation of Mg2+-dependent phosphatidate phosphohydrolase. AB - Adipose cytosol treated with spermine showed an aggregation of a cytosolic component which was isolated by centrifugation at 16,000 X g for 20 min. The resultant pellet contained 10% of protein, 40% of lipid and over 75-97% of Mg2+ dependent phosphatidate phosphohydrolase and CTP:phosphocholine cytidylyltransferase activities present in the original cytosol. The specific activities of these enzymes increased 4-fold by the spermine treatment. Characterization of lipids in this component indicated the presence of mainly phospholipids. These studies suggest that the interaction between spermine, the cytosolic component and microsomal membranes may be involved in the translocation of Mg2+-dependent phosphatidate phosphohydrolase. PMID- 3026489 TI - Transformation of 15-hydroperoxyeicosapentaenoic acid to lipoxin A5 and B5, mono- and dihydroxyeicosapentaenoic acids by porcine leukocytes. AB - 15-Hydroperoxy[1-14C]eicosapentaenoic acid derived from eicosapentaenoic acid (EPA) was incubated with suspensions of porcine leukocytes. Incubation with porcine leukocytes resulted in the formation of seven dihydroxy compounds, one monohydroxy and one hydroxyepoxy compound. After separation by reverse-phase and straight-phase HPLC, GC/MS analysis revealed that these metabolites were four isomers of 8,15-diHEPEs, two isomers of 14,15-diHEPEs, one isomer of 5,15-diHEPE, 15-HEPE and an epoxyalcohol: 13-hydroxy-14,15-epoxyeicosatetraenoic acid. In addition to the above metabolites, two trihydroxytetraene derivatives were also isolated. GC/MS and ultraviolet spectroscopy identified the two trihydroxypentaene derivatives as 5,6,15-trihydroxy-7,9,11,13,17-eicosapentaenoic acid (lipoxin A5) and 5,14,15-trihydroxy-6,8,10,12,17-eicosapentaenoic acid (lipoxin B5). This study demonstrated that the 15-hydroperoxide of EPA can be actively converted to various hydroxylated products via the 5-, 12- and 15 lipoxygenase as well as epoxyisomerase pathways in the porcine leukocytes. PMID- 3026490 TI - Activation of leukocyte movement and displacement of [3H]leukotriene B4 from leukocyte membrane preparations by (12R)- and (12S)-hydroxyeicosatetraenoic acid. AB - Both (12R)- and (12S)-hydroxyeicosatetraenoic acid were demonstrated to produce aggregation of rat leukocytes and enhance human leukocyte chemokinesis. (12R) Hydroxyeicosatetraenoic acid was 10-20-fold more potent than (12S) hydroxyeicosatetraenoic acid but at least 500-fold less potent than leukotriene B4 in these assays. These relative potencies are correlated with the potencies of (12R)- and (12S)-hydroxyeicosatetraenoic acid for competition of [3H]leukotriene B4 binding to rat and human leukocyte membrane preparations. PMID- 3026491 TI - Affinity chromatography and some properties of the angiotensin-converting enzyme from human heart. AB - Human heart angiotensin-converting enzyme has been purified by affinity chromatography on immobilized N-[1(S)-carboxy-5-aminopentyl]-Gly-Gly. The isolation procedure permitted a 1650-fold-purified enzyme to be obtained. The specific activity of angiotensin-converting enzyme was 38 units per mg protein. The molecular weight of enzyme determined by polyacrylamide gel electrophoresis in denaturing conditions was 150,000. The isoelectric point (5.3) of the enzyme was determined by chromatofocusing. The Km values of the enzyme for Bz-Gly-His Leu and angiotensin I are 1.2 mM and 10 microM, respectively. Substrate inhibition of heart angiotensin-converting enzyme with a K's of 14 mM has been shown. The human heart enzyme is inhibited by SQ 20881 (IC50 = 40 nM). It was shown that NaCl, CaCl2 as well as Na2SO4 in the absence of Cl- are activators of the heart angiotensin-converting enzyme, whereas CH3COONa and NaNO3 have no effect on a catalytic activity of the enzyme. PMID- 3026492 TI - Partial purification and characterization of an aldohexose 1-P phosphatase from pig skeletal muscle. AB - An enzyme with a molecular weight of 54,000 which possesses phosphatase activity acting on glucose 1-P, galactose 1-P and mannose 1-P has been partially purified and characterized from pig skeletal muscle. The enzyme is free of phosphoglucomutase and galactokinase activities, and it possesses a neutral optimum pH. Pi acts as an inhibitor; glucose, galactose and mannose do not produce any effect. Divalent cations are required for activity, Mg2+ being the most effective activator. Micromolar levels of fluoride and millimolar levels of chloride act as inhibitors; however, vanadate does not produce any effect. The enzyme may have an important role when galactose accumulates in tissues; for example, in galactosemic patients and in young animals ingesting high-galactose diets. PMID- 3026494 TI - ["Freezing" of the conformation of thin filaments in a single skinned fiber of the rabbit muscle by glutaraldehyde]. AB - It is shown that short treatment of a single skinned rigor fibre from rabbit m X psoas with 0.05% glutaraldehyde in the absence of Ca ions leads to a modified state of the contractile apparatus. After the addition of 5 mM MgATP in the absence of Ca ions to the fibre a sharp rise and subsequent slow decay of tension were observed in contrast to the tension drop in case of the control (unmodified) specimen. The tension transients following quick stretch (L 0.5%) were similar to those for Ca-activated tension. In case of the modified relaxed fibre such a phenomenon was not observed. These results can be explained by "freezing" with glutaraldehyde the thin filament structure either in the "on" or "off" states. The relation of these results to the cooperativity in the regulation mechanism of contraction is discussed. PMID- 3026493 TI - Nucleoside-triphosphatase and hydrolysis of thiamin triphosphate in Escherichia coli. AB - A membrane-bound nonspecific triphosphatase of E. coli was solubilized and purified to a homogeneous SDS-acrylamide gel electrophoresis band. It was found to be a single polypeptide of 16 kDa requiring no Mg2+, with an optimal pH at 6.5. The substrate specificity was broad and a nonspecific Mg2+-independent ribonucleoside-triphosphatase (NTPase) activity was expressed together with thiamin-triphosphatase activity. The molecular size and characteristics were clearly different from the known NTPase (EC 3.6.1.15). Using the purified thiamin triphosphatase II, ATP:thiamin-diphosphate phosphoryl transferase (EC 2.7.4.15) activity was demonstrated with an optimal pH of approx. 5.3. Considering its kinetic parameters and other characteristics, however, the thiamin triphosphate synthesizing activity was not thought to take part in cellular thiamin triphosphate synthesis. The possibility that thiamin-triphosphatase II plays a part in the hydrolysis of thiamin triphosphate to control its cellular level is suggested. PMID- 3026495 TI - [Transformation of ferromagnetic suspensions in the animal body]. AB - Ferromagnetic suspension (FS) was introduced into rat and mouse organisms by different ways. Transformation of FS into some organs was estimated by ESR-method within the temperature region 80-250 K. It was shown that FS introduced in the animal organism was utilized in it very quickly. PMID- 3026496 TI - [Computer analysis of anisotropic ESR spectra and the structure of radical H and OH adducts of nucleic acid components in frozen aqueous solutions]. AB - Computer analysis of ESR spectra recorded under gamma-irradiation of polycrystal and glass-like solutions of nucleotides and nitrous bases was carried out to determine the localization sites of free valency (damage) in DNA macromolecule when interacting with water radiolysis products (OH and H). The analysis was performed on a mini-computer with the program of reconstruction of ESR anisotropic spectra in the variation search regime. When calculating the theoretical spectra anisotropic width of the lines, g-factors and HHS constants on X and Z axes were taken into account. Good reproduction of the experimental spectra on the computer was achieved, as a result the ESR spectra were obtained. Relative contribution of each of OH- and H-adducts of nitrous bases to the total ESR spectrum of gamma-irradiated DNA will be determined in future. PMID- 3026497 TI - [1H-NMR study of the peptide corresponding to the ACTH-like sequence of the variable region of human G1 Eu heavy chain immunoglobin]. AB - Synthetic decapeptide corresponding to ACTH-like sequence of the variable part of the heavy chain of immunoglobulin G1 Eu was studied by two-dimensional 1H-NMR spectroscopy (400 MHz). A complete assignment of signals in the peptide spectrum was made. The decapeptide was shown not to have any ordered spatial structure, and was characterized by a high extent of flexibility of the oligopeptide chain, except for the peptide bond with an N-terminal residue. PMID- 3026499 TI - [Isolation and molecular kinetic properties of the angiotensin-converting enzyme from the human heart]. AB - An angiotensin-converting enzyme was isolated from human heart using N[-1(S) carboxy-5-aminopentyl]glycyl-glycine as an affinity adsorbent. The isolation procedure resulted in an enzyme purified 1650-fold. The enzyme specific activity was 38.0 u./mg protein, Mr = 150 kD. The pH optimum for the angiotensin converting enzyme towards Hip-His-Leu lies at 7.8, Km = 1.2 mM. The enzyme was inhibited by the substrate (Ks' = 14 mM). The enzyme effectively catalyzed the hydrolysis of angiotensin I (Km = 10 microM; kcat = 250 s-1). NaCl, CaCl2 as well as Na2SO4 in the absence of Cl- activated the enzyme, whereas CH3COONa and NaNO3 did not influence the enzyme activity. It was found that the bradykinin potentiating factor inhibited the cardiac angiotensin-converting enzyme with IC50 = 4.0 X 10(-8) M. PMID- 3026498 TI - [Glutamate dehydrogenase--an activator of NAD-kinase in the rabbit liver]. AB - An electrophoretically homogeneous preparation of a NAD-kinase activator from rabbit liver was obtained and its physico-chemical properties were investigated. The molecular mass of the monomer and oligomer, pI, number of SH-groups per enzyme subunit and some other factors were determined. The similarity of activator properties to those of glutamate dehydrogenase and the revealed glutamate dehydrogenase activity of the NAD-kinase activator permitted to identify the latter as glutamate dehydrogenase. It was demonstrated that the enzyme activates NAD-kinase 2-4 times already at the glutamate dehydrogenase: NAD kinase ratio of 2:1. The effect of glutamate dehydrogenase on the enzyme consists in an increase of Vmax; the KmNAD value for the NAD-kinase reaction remains thereby unchanged. The physiological role of the interaction between the two enzymes is discussed. PMID- 3026501 TI - [A three-cycle mechanism of the respiratory chain]. AB - A scheme of the respiratory chain is presented, according to which three delta mu H-generators (complexes I, III and IV) provide for the transmembrane transport of ten H+ ions per atom of adsorbed O2. It is assumed that all the three delta mu H generators operate in accordance with the same mechanism, namely, they translocate electrons at a distance averaging 1/2 of membrane width, whereas protons moving in the opposite direction pass the other halfwidth across the membrane. A redox cycle functions in each of the three sites of the energy coupling mechanism: the flavin (F) cycle in complex I, the Q-cycle in complex III and the O-cycle in complex IV. These cycles are interconnected by mobile carriers: the F- and Q-cycles by ubiquinone and the Q- and O-cycles by cytochrome c. PMID- 3026500 TI - [Inhibition of platelet aggregation by dinitrosyl iron complexes with low molecular weight ligands]. AB - The dinitrosyl iron complexes (DNIC) with thiosulphate, cysteine or phosphate were shown to inhibit in vitro (in citrate plasma) the human platelet aggregation induced by ADP, collagen or adrenaline. This effect cannot be explained by the toxic action of DNIC on the platelet membrane, since DNIC-pretreated platelets are capable of aggregating under the action of 10(-8) M/ml of phorbol ester, which is known to cause direct activation of protein kinase C. The antiaggregatory activity of DNIC exceeds that of Na-nitroprusside and seems to be due to nitric oxide capable to activate guanylate cyclase of platelets. Using the EPR method, it was shown that addition of DNIC to platelet-enriched plasma results in a rapid transfer of Fe(NO)2 groups to the coupled RS(-)-groups proteins of plasma and, apparently, of platelet membrane proteins. These protein DNIC seem to be the source of NO which inhibits human platelet aggregation. PMID- 3026502 TI - Ca2+-sensitive cross-bridge dissociation in the presence of magnesium pyrophosphate in skinned rabbit psoas fibers. AB - We find that at 6 degrees C in the presence of 4 mM MgPPi, at low or moderate ionic strength, skinned rabbit psoas fibers exhibit a stiffness and an equatorial x-ray diffraction pattern similar to that of rigor fibers. As the ionic strength is increased in the absence of Ca2+, both the stiffness and the equatorial x-ray diffraction pattern approach those of the relaxed state. This suggests that, as in solution, increasing ionic strength weakens the affinity of myosin cross bridges for actin, which results in a decrease in the number of cross-bridges attached. The effect is Ca2+-sensitive. Assuming that stiffness is a measure of the number of cross-bridge heads attached, in the absence of Ca2+, the fraction of attached cross-bridge heads varies from approximately 75% to approximately 25% over an ionic strength range where ionic strength in solution weakens the binding constant for myosin subfragment-1 binding to unregulated actin by less than a factor of 3. Therefore, this phenomenon appears similar to the cooperative Ca2+ sensitive binding of S1 to regulated actin in solution (Greene, L. E., and E. Eisenberg, 1980, Proc. Natl. Acad. Sci. USA, 77:2616). By comparing the binding constants in solution and in the fiber under similar conditions, we find that the "effective actin concentration," that is, the concentration that gives the same fraction of S1 molecules bound to actin in solution as cross-bridge heads are bound to actin in a fiber, is in the millimolar range. An effective actin concentration in the millimolar range suggests that the strength of actin binding to cross-bridges in fibers may be several orders of magnitude weaker than the strength of ATP binding. Previously, it has been assumed that these two quantities were equal, as this gives the minimum energy loss when ATP dissociates the cross-bridge from actin (Morales, 1980, J. Supramol. Struct., 3:105:1975; Eisenberg, E.,Hill, T. L. and Y. Chen, 1980, Biophys. J., 29:195). PMID- 3026503 TI - Measurement of rotational molecular motion by time-resolved saturation transfer electron paramagnetic resonance. AB - We have used saturation-recovery electron paramagnetic resonance (SR-EPR), a time resolved saturation transfer EPR technique, to measure directly the microsecond rotational diffusion of spin-labeled proteins. SR-EPR uses an intense microwave pulse to saturate a spin population having narrow distribution of orientations with respect to the magnetic field. The time evolution of the signal is then observed. The signal increases in time as saturation is relieved by spin-lattice relaxation (Tl) as well as by saturation transfer due to spectral diffusion (Tsd), which is a function of rotational diffusion (Tr) and spectral position. In the presence of both events, the recovery is biphasic, with the initial phase related to both Tr and Tl, and the second phase determined only by Tl. We have measured the saturation recoveries of spin-labeled hemoglobin tumbling in media of known viscosities as a function of rotational correlation time (Tr) and pulse duration (tp). The Tr values estimated from the initial phase of recovery were in good agreement with theory. Variation of the pulse time can also be used to determine Tr. For tp less than Tsd, the recoveries were observed to be biphasic, for tp greater than Tsd a single-exponential. T1 values were determined from the recoveries after pulses quenching spectral diffusion or from the second phase of recovery after shorter pulses. These results demonstrate that SR-EPR is applicable to the study of motion of spin-labeled proteins. Its time resolution should provide a significant advantage over steady state techniques, particularly in the case of motional anisotropy or system heterogeneity. PMID- 3026504 TI - Determination of relative spin concentration in some high-spin ferric proteins using E/D-distribution in electron paramagnetic resonance simulations. AB - Lineshape simulations are presented for the multiple, overlapping X-band electron paramagnetic resonance (EPR) spectra in two non-heme, high-spin iron proteins: phenylalanine hydroxylase (PAH) and diferric transferrin. The aim of the calculations is to determine the fraction of iron contributing to each of the sites visible by EPR. The simulations are limited to the experimentally accessible transitions occurring at g-values greater than 1.7. In both PAH and transferrin, at least one of the iron sites is characterized by the ratio of zero field splitting parameters, E/D, near 1/3 and a broad, asymmetric lineshape. A distribution in E/D-values is used in the simulations to account for this breadth and asymmetry. To test the E/D-distribution model, experimental X-band spectra of diferric transferrin at several salt concentrations are fit by simulation. In this test, first the low-field features arising from transitions between the lowest Kramers doublet levels are simulated using E/D-distributions for two sites. Second, parameters that provide a good fit for the lowest doublet transitions are shown also to fit the resonance near an effective g-value of 4.3 from the middle Kramers doublet transition. When applied to spectra of PAH in the resting state, the E/D-distribution approach accounts for the intensity of one of the two major species of iron. The other species is characterized by E/D = 0.032, and the spectrum of this portion of the resting enzyme may be simulated using a frequency-swept Gaussian lineshape. Spectra for the enzyme in an inhibitor saturated state are also simulated. The simulations are consistent with previous biochemical studies that indicate that only the E/D = 0.032 form of iron participates in catalysis. PMID- 3026505 TI - Application of the theory of enzyme subunit interactions to ATP-hydrolyzing enzymes. The case of Na,K-ATPase. AB - The theory developed by T. L. Hill (1977, Proc. Natl. Acad. Sci. USA, 74:3632 3636) for enzyme interactions is applied to a dimeric enzyme, the subunits of which may each exist in three distinct states (as in a uni-bi kinetic mechanism). It is shown that when simultaneous binding of substrate to both subunits is excluded, the complex kinetic mechanism of the dimer reduces to a simpler scheme with two distinct, but analogous, cycles that are in principle separately observable in kinetic experiments. Because of the intersubunit interactions, which are explicitly taken into account, the two cycles have different Michaelis constants and maximal velocities. The model exhibits negative cooperativity and enhanced reactivity, relative to a monomeric enzyme. The theory is applied to Na,K-ATPase for which a complete, bicyclic, kinetic mechanism and rate constants are available. When taken together with other evidence, structural as well as functional, the striking similarity of the observed kinetics with that developed for a dimeric enzyme strongly suggests that the functional unit of Na,K-ATPase is a dimer. The free energy differences (calculated from the known rate constants) between intermediates are 6-16 kJ/mol, comparable, for example, to the free energy associated with the formation of a base pair in a nucleic acid double helix. The possible relevance of these results for other ATPases is briefly discussed. PMID- 3026506 TI - Synthesis of fragments by classical solution methods for use in cytochrome c semisynthesis. PMID- 3026507 TI - The evolution of eukaryotic ribosomal DNA. AB - Mutations occur randomly throughout the ribosomal DNA (rDNA) sequence. Molecular drive (unequal crossing-over, gene conversion, and transposition) spreads these variations through the multiple copies of rDNA. Forces of selection act upon the variants to favor and fix them or disfavor and eliminate them. Selection has not permitted changes in regions within rRNA vital for its function; these sequences are evolutionarily conserved between diverse species. Possible functions for some of these conserved sequences are discussed. The secondary structure of rRNA is also highly conserved during evolution. However, eukaryotic rRNA is larger than prokaryotic rRNA due to blocks of "expansion segments". Arguments are put forward that expansion segments might not play any functional role. Other examples are reviewed of rDNA sequence insertion or deletion, including introns and the internal transcribed spacer 2. PMID- 3026508 TI - Suitability of oligopeptides for induction of hormonal imprinting--implications on receptor and hormone evolution. AB - Hormonal imprinting induced in Tetrahymena and in Chang liver cells with di-, tri , tetra- and pentapeptides (synthetic opioids and their fragments) has shown that both cell types are able to differentiate the related molecules from one another. The dipeptide phenylalanine + proline induced a measurable imprinting in the liver cells, and chain length increase, especially terminal coupling with tyrosine enhanced the imprinting potential enormously. Intra-chain changes in the amino acid sequence had a measurable effect on the intensity of imprinting. The molecules showing the relatively strongest physiological action accounted for the most intensive imprinting in both cell types; this indicates that, in all probability, induction of binding site formation plays a key role in the development of signal molecules, and thereby in hormone evolution. PMID- 3026509 TI - [Electrophysiological research on the coupling of excitation-contraction during alpha-adrenoreceptor activation in blood vessel smooth muscles]. AB - Alpha 1-adrenoceptor blocker--prazosin--was found to inhibit noradrenaline induced depolarization and concentration in the smooth muscles of the portal rabbit vein, indicating that this reaction was due to alpha 1-adrenoceptor activation. In the pulmonary artery both alpha 1 and alpha 2-adrenoceptors appear to be involved in noradrenaline excitatory action, as the effect was not completely inhibited by prazosin. The results suggest that hypotensive action of prazosin is related to the cessation of Ca2+ ion influx through alpha 1-operated calcium channels. The decrease in Ca2+ influx through voltage-dependent calcium channels due to prazosin-evoked elimination of depolarization can also contribute to this effect. PMID- 3026510 TI - [Formation of the superoxide anion radical during the oxidation of biogenic amines catalyzed by mitochondrial monoamine oxidase]. AB - The studies on the activity of monoamine oxidase from human placenta, using 2 phenylethylamine as a substrate, corroborate the hypothesis on the possible superoxide radical generation upon FAD oxidation at the second (aerobic) stage of monoamine oxidase reaction. It has been shown that hydrogen peroxide, but not other activated O2 forms, was the end product of this reaction. No superoxide radical generation took place in such systems. And therefore, the induction of lipid peroxidation in the presence of catalase was impossible in mitochondrial membranes containing monoamine oxidase and amines oxidized by it. PMID- 3026512 TI - [Action of triftazin on the physicochemical properties and the membrane proton pump of the synaptic vesicles in the rat brain]. AB - The effects of trifluoperazine (TFP) on some membrane processes were studied in the isolated rat brain synaptic vesicles (SV). TFP (10(-5)-10(-4) M) was found to cause a sharp rise in the intensity of light scattering by SV suspension which was due both to an increased vesicle aggregation and to changes in the refraction index of the membrane. In addition, TFP blocked the ATP-dependent proton transport into the vesicles (K0.5 = 10(-6) M) with the concomitant stimulation of the ATPase activity which suggests an uncoupling effect caused by the permeation of this weak base through the membrane and subsequent protonation in an acid interior medium resulting in the elimination of a proton gradient. Thus, the neuroleptic drug--TFP has various effects on membrane processes which are apparently unrelated to its recognized role as a calmodulin antagonist. PMID- 3026511 TI - [Effect of thiol-specific reagents on the activity of PaeI and PaeII restrictases from Pseudomonas aeruginosa]. AB - 5,5'-dithiobis-2-nitrobenzoic acid, N-ethylmaleimide, and parachloromercuribenzoate have been demonstrated to inhibit the activity of restrictases PaeI and PaeII from Ps. aeruginosa bacterial cells. Restrictase PaeII was more sensitive to the action of thiol-specific reagents, as compared to PaeI. The minimal concentration of reagents for SH-groups that completely inhibited the activity of restrictases PaeI and PaeII was determined. The protective effect against the inhibitory action of 5,5'-dithiobis-2-nitrobenzoic acid on the activity of PaeII was observed after preincubation of these enzymes with phage lambda DNA and Mg2+ cations. It is suggested that restrictase PaeI and PaeII molecules contain SH-groups, essential for the enzymatic activity. They are believed responsible for restrictase binding with DNA substrate. PMID- 3026513 TI - [Effect of fenazepam on the ACTH content of the blood plasma in inbred mice under stress exposure]. AB - The influence of phenazepam on plasma ACTH level before and after exposure to stress in the "open field" test was investigated in C57BL/6, BALB/c and F1(C57BL/6 X BALB/c) mice. The differences in the dose-dependent drug effect on the hormone level between strains were studied. A correlation between ACTH level and a characteristic animal behaviour has been established for different strains. PMID- 3026514 TI - [Effect of beta-endorphin and myelopeptides on the cAMP level and proliferation of lymphocytes in vitro]. AB - Beta-endorphin (10(-11)-10(-9) M) has been shown to induce naloxone-independent depression of the proliferative activity of human peripheral lymphocytes (HL), stimulated by pokeweed mitogen without affecting PHA-stimulated HL proliferation. Beta-endorphin (10(-10)-10(-7) M) also caused changes in HL cAMP level, that were blocked by naloxone. Marked individual sensitivity to beta-endorphin effects has been noted. It has been also shown that a bone marrow preparation, stimulating antibody production (myelopeptides), causes naloxone-independent depression in the proliferative activity of HL, stimulated by PHA and pokeweed mitogen, as well as naloxone-blocked decrease in cAMP HL level. It has been concluded that beta endorphin interacts with several types of opiate lymphocyte receptors and that opioids, contained in myelopeptides, are involved in the realization of myelopeptide effect on lymphocytes. PMID- 3026515 TI - [Morphofunctional changes in the microcirculatory bed of the synovial membrane in antigen-induced arthritis]. AB - Ultrastructural changes of synovial capillary and venule wall have been investigated in rabbits with antigen-induced arthritis, with the permeability of microcirculatory bed determined using 99mTc pertechnetate. The correlation between the incidence of ultrastructural microvascular damages in exudative phase of arthritis and the increased permeability has been established. Ultrastructural changes occurring in proliferative phase of arthritis are considered as adaptive to conditions of increased permeability. PMID- 3026516 TI - [Effect of polypeptides isolated from the heart on the course of experimental myocardial infarct]. AB - Polypeptides excreted from the heart have been shown to exert a positive influence on myocardial infarction caused by izadrin injection in experimental rats. PMID- 3026517 TI - [Effect of corticotropin on the rate of 32P-orthophosphate incorporation into the synaptosome phosphoinositides of the ischemic rat brain]. AB - A high rate of 32P turnover in polyphosphoinositides (up to 80% of the total radioactivity) was found in synaptosomes of normal and ischemic rat brain. Corticotropin (ACTH) increases the rate of 32P turnover in polyphosphoinositides of normal synaptosomes and decreases it in ischemic synaptosomes. Depolarization (high KCl concentration in the incubation medium) activates polyphosphoinositide metabolism in normal (by 50%) and ischemic (by 30%) synaptosomes. The combined influence of depolarization and ACTH results in the additive effect. Thus, a stimulating effect of ACTH on phosphoinositide metabolism disturbed in ischemia was recovered during depolarization of ischemic synaptosomes. PMID- 3026518 TI - [Induction of blood plasma carboxycathepsin (angiotensin I-converting enzyme) in normo- and hypertensive rats in response to a single administration of captopril]. AB - The experiments were carried out to elucidate the effect of carboxycathepsin (CC) activity inhibition by a specific inhibitor--captopril--on plasma enzyme concentration in normotensive rats and rats with renovascular hypertension. A single oral administration of captopril (10 mg/kg body weight) produced an increase in CC concentration in both hypertensive and sodium-depleted normotensive rats with a parallel decrease in arterial pressure, but had no effect on sodium-repleted normotensive rats. It is suggested that the increase in plasma CC concentration is a compensatory response to the inhibition of CC activity by captopril; it is also possible that the increase observed reflects the state of renin-angiotensin system. PMID- 3026519 TI - [Analysis of the specific 3H-diazepam binding in the brain of inbred mice with differing emotionality]. AB - The influence of sodium chloride and gamma-aminobutyric acid (GABA) on H3 diazepam binding in CNS membrane fraction were investigated in C57Bl/6, BALB/c mice and their F1 hybrids. The addition of GABA (1, 10 and 100 microM) elevated the level of radioligand binding with CNS membranes in all the inbred mice in similar manner. The higher stimulating effect of sodium chloride (50, 100 and 150 mM) on the H3-diazepam reception was found in BALB/c mice and F1 hybrids, as compared to C57Bl/6 mice. It has been suggested that membrane-dependent conformational reconstructions of supramolecular receptor complex are the cause of genetic differences in the regulation of H3-diazepam reception by Cl(-) ionophore. PMID- 3026521 TI - [Regulation of 3H-dopamine release by presynaptic GABA and glutamate heteroreceptors in the synaptosomes of the rat nucleus accumbens]. AB - The influence of GABA, muscimol, delta-aminolevulinic acid (DALA), baclofen and L glutamate on K+-evoked release of 3H-dopamine (3H-DA) from the rat brain n. accumbens crude synaptosomal fraction was studied in superfusion experimental conditions. Both GABA-receptor agonists--GABA and muscimol (50 microM) depressed the 3H-DA release by bicuculline- and picrotoxin-sensitive manner. On the contrary, glutamate, DALA and baclofen led to the increase in 3H-DA efflux independently of the presence of GABA-receptor antagonists. While the action of glutamate was antagonized by glutamate-receptor blocker--glutamic acid diethyl ester (GDEE), the effects of DALA and baclofen were suppressed upon adding to superfusion medium of GABA uptake inhibitors (nipecotic acid and 2,4 diaminobutyric acid) but not GDEE. The data obtained demonstrate that 3-H-DA secretion from n. accumbens is inhibited by GABA- and stimulated by glutamate heteroreceptors. At the same time the mechanism of interaction between baclofen, DALA and GABA-uptake blockers effects with 3H-DA release needs special investigations. PMID- 3026520 TI - [Role of the GABA- and cholinergic systems in the formation of a dissociated state to an antioxidant of the 3-hydroxypyridine class]. AB - The experiments on rats have shown that the elaboration of conditioned drinking reflex in T-maze during administration of 2-ethyl-6-methyl-3-hydroxypyridine antioxidant with an anti-stress effect was accompanied by the development of state dependent learning. However, its formation was slower, as compared to state dependent learning in response to the known psychotropic drugs. The replacing test with the injection of bicuculline, picrotoxin, Ca valproate, Ro-15-1788, benactyzine, Cleregil, etc. during state dependent learning made it possible to establish the role of GABA and cholinergic systems in the formation of state dependent learning and in the development of disorders in emotional behavioural reactions after long-term administration and withdrawal of 3-hydroxypyridine. PMID- 3026522 TI - A monoclonal antibody detecting a novel antigen expressed in the HTLV-I-infected cells. AB - A monoclonal antibody, FTF 148, was prepared by hybridizing murine myelomal cells (NS-1) and spleen cells of BALB/c mice immunized with cultured cells derived from an adult T cell leukemia (ATL) patient (KUT-2 cells). This monoclonal antibody reacted with all of the human T cell leukemia virus I (HTLV-I)-infected cell lines tested but did not react with other T cell lines derived from acute lymphocytic leukemia, Epstein-Barr virus-transformed B cell lines, or an erythroleukemic cell line. This monoclonal antibody was not directed to viral antigens because it reacted equally well with almost all KUT-2 and MT-1 cells, only 1% to 3% of which were ATL-associated antigen-positive. In contrast to interleukin 2 receptors expressed on both ATL cells and normal phytohemagglutinin stimulated blasts, this antigen was not expressed on the latter cells. The antigen, mainly expressed on the cell membrane, was analyzed by metabolic labeling with 3H-leucine and surface labeling with 125I followed by cell lysis and immunoprecipitation with the FTF 148 antibody. The findings obtained by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that p50 and p74 proteins were specifically precipitated and the antigen was also different from the product of the Xs gene of HTLV-I. PMID- 3026523 TI - Spectrin is associated with membrane-bound actin filaments in platelets and is hydrolyzed by the Ca2+-dependent protease during platelet activation. AB - We recently showed that platelets contain submembranous actin filaments that are linked to glycoprotein (GP) Ib on the plasma membrane. In the present study, experiments were performed to determine whether spectrin was associated with these filaments. The membrane-bound filaments were isolated from Triton X-100 (Sigma, St Louis) lysates of unstimulated platelets by differential centrifugation. Platelet spectrin was detected immunologically by using antibodies against human brain and RBC spectrin. Immunoblots showed that platelet spectrin consisted of two polypeptides (mol wt 240,000 and 235,000) that were similar in apparent mol wt to those of the alpha and beta chains of brain spectrin but differed slightly from those of RBC spectrin (mol wt 240,000 and 220,000). Immunoprecipitation experiments identified platelet spectrin as two minor polypeptides migrating on sodium dodecyl sulfate (SDS)-polyacrylamide gels between actin-binding protein (mol wt 250,000) and the platelet polypeptide P235 (mol wt 235,000). Immunoblots of fractions isolated from Triton X-100-lysed platelets revealed that the alpha and beta chains of platelet spectrin were associated almost entirely with the actin filaments that were linked to the plasma membrane. Little spectrin was recovered in the Triton X-100-soluble fraction or with the actin filaments that were not membrane bound. During activation of platelets with thrombin or ionophore A23187, the alpha and beta chains of spectrin were hydrolyzed, generating a major degradation product of mol wt 160,000 and a minor one of mol wt 170,000. These two hydrolytic products were also generated in Triton X-100 lysates incubated in the presence of Ca2+ but were not produced when lysates were treated with leupeptin, ethylene glycol bis(beta aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA), or N-ethylmaleimide, known inhibitors of the Ca2+-dependent protease. These experiments show that spectrin is a previously unidentified component of the membrane-bound actin filament network and that hydrolysis of spectrin by the Ca2+-dependent protease may regulate the interactions of the filaments during platelet activation. PMID- 3026524 TI - Association between gelatinase release and increased plasma membrane expression of the Mo1 glycoprotein. AB - Mo1, a glycoprotein heterodimer (gp 155,95) that functions as an adhesion promoting molecule and as the C3bi receptor of human myeloid cells, is expressed in increased amounts in the plasma membrane after exposure of polymorphonuclear leukocytes (PMNs) to various stimuli. Previous studies have suggested that secondary granules represent an intracellular pool of Mo1 that, upon degranulation, fuse with the plasma membrane resulting in a tenfold increase in surface expression of Mo1. To determine the intracellular location of Mo1, we monitored Mo1 expression by immunofluorescence and compared it to the release of myeloperoxidase (MPO, a marker for the primary granules), vitamin B12 binding protein (B12BP, secondary granules), and gelatinase (gelatinase-containing organelles) following exposure to various stimuli. Human neutrophils stimulated with 20 mmol/L fluoride for 16 minutes exhibited a twofold increase in Mo1 expression and gelatinase release but no enhanced release of primary or secondary granular contents. In a similar fashion, incubation of cells at 37 degrees C for five minutes with 7.5 X 10(-9) to 10(-6) mol/L N-formyl-methionyl-leucyl phenylalanine (FMLP) resulted in significant increases in both surface Mo1 expression (three- to fivefold) and gelatinase release (five- to eightfold) without significant release of either MPO or B12BP. In addition, both the fluoride and FMLP experiments demonstrated that Mo1 up-modulation alone is not sufficient to activate superoxide (O2-) production. These data indicate that at least one intracellular storage pool of Mo1 is the gelatinase-containing organelles and that their fusion with the plasma membrane results in increased expression of Mo1 on the cell surface. PMID- 3026525 TI - Synergistic action of two murine monoclonal antibodies that inhibit ADP-induced platelet aggregation without blocking fibrinogen binding. AB - We have used two murine monoclonal antibodies, each directed against one component of the human platelet membrane glycoprotein (GP) IIb-IIIa complex, to examine further the molecular requirements for fibrinogen binding to the platelet surface and subsequent platelet-platelet cohesion (aggregation). Although neither AP3, which is directed against GPIIIa, nor Tab, which is specific for GPIIb, were individually able to inhibit adenosine diphosphate (ADP)-induced fibrinogen binding, platelet aggregation, or secretion, the combination of AP3 and Tab completely abolished platelet aggregation and the release reaction. Unexpectedly, this synergistic inhibition of platelet-platelet cohesion occurred in the presence of apparently normal fibrinogen binding. Both the number of fibrinogen molecules bound and the dissociation constant for fibrinogen binding remained essentially unchanged in the presence of these two antibodies. Inhibition of aggregation was dependent upon the divalency of both AP3 and Tab because substitution of Fab fragments of either antibody for the intact IgG resulted in a complete restoration of both aggregation and secretion. In contrast to ADP induction, thrombin-activated platelets neither aggregated nor bound fibrinogen in the presence of AP3 plus Tab but were fully capable of secretion, which illustrated the multiple mechanisms by which the platelet surface can respond to different agonists. These data demonstrate that fibrinogen binding to the platelet surface alone is not sufficient to support platelet-platelet cohesion and that an additional post-fibrinogen-binding event(s) that is inhibitable by these two monoclonal antibodies may be required. PMID- 3026526 TI - The importance of SH-containing substances for red blood cells in acute myeloic leukemia. AB - Electron spin resonance (ESR) spectra of lyophilized erythrocytes obtained from patients with acute myeloid leukemia (AML) show, in comparison to controls, a characteristic change especially in the low-field region of the spectrum concomitant with a reduction of the spin concentration. This effect can be simulated by addition of SH-containing substances (e.g. reduced glutathione or cysteine) to healthy erythrocytes. S-S containing compounds exhibit no effect. Since SH-containing substances can hardly permeate plasma membranes, the membrane surface seems to be defective in the case of "AML" erythrocytes. Furthermore, it can be concluded that the concentration of SH-containing substances, such as cysteine, is increased in the plasma of AML-patients, which could be confirmed by HPLC-measurements. In the case of a successful treatment of the patients with alexan, daunoblastin, and thioguanine the spin concentration increased again and the resulting ESR spectrum is very similar to the control spectrum. It should be pointed out, that the ascorbic acid concentration is very low in both plasma and erythrocytes of AML patients. PMID- 3026527 TI - Lymphocytic plasmocytoid lymphoma with a three-banded gammopathy: reactivity of one of these paraproteins with cytomegalovirus. AB - We report the case of a 70-year-old woman suffering from small lymphocytic, plasmocytoid lymphoma with abdominal lymphomas and infiltration of the lung and the bone marrow. A three-banded, IgG lambda, IgM lambda and IgA lambda paraproteinemia was determined using immunofixation. Because of the patient's high antibody titre against cytomegalovirus (CMV), the possible reactivity of these paraproteins with CMV was studied. The immunoglobulins were transferred to nitrocellulose sheets by a contact diffusion blotting system. CMV was applied to these sheets and the IgG lambda-paraprotein was shown to bind CMV. The reactivity of only one of the paraproteins with CMV suggests an oligoclonal origin of this gammopathy. In addition to the malignant disease an abnormal immune response to a CMV infection could be the cause of this three-banded gammopathy. PMID- 3026528 TI - Metal-induced conformational changes in calmodulin. PMID- 3026529 TI - Airflow limitation, asbestos exposure and lung size. PMID- 3026530 TI - A morphometric study of subcellular responses in frog pituitary corticotropic cells to constant environmental conditions. AB - Ultrastructural morphometry (point-counting method) was used to evaluate stereological parameters of rough endoplasmic reticulum (RER), Golgi complex, secretory granules, mitochondria and lysosomes of Rana ridibunda pituitary corticotropic (ACTH) cells in animals subjected to constant environmental conditions (CEC) in relation to temperature and photoperiod. During this experiment, corticotropic cells are particularly sensitive to the effects of changes in environmental conditions in such a way that they course through a phase of RER and Golgi complex hypertrophy after 7 days on CEC and a phase of enhancement in the amount of secretory granules (the 11th day). After 15 days on CEC, the cells seemingly recuperate almost all the control evaluate parameters. These results strongly suggest that, in frog kept in captivity, ACTH cells play an important role in the acclimation process to new constant environmental conditions. PMID- 3026531 TI - p53 and transformation by SV40. AB - The large T antigen of SV40 is able to immortalize and transform primary and established cells in culture, and can, at least in certain cases, confer a tumorigenic phenotype on the infected cell. T antigen has been shown to induce cellular DNA synthesis in the infected cell and this activity is likely to be instrumental in T antigen mediated oncogenesis. A property of T antigen which may be of paramount importance to its oncogenic and mitogenic activities is its ability to specifically bind and stabilize the cellular protein p53. p53 has been implicated in the control of the passage of the cell from G0 arrest to G1 and S phase. Furthermore, altered p53 expression is strongly associated with various phenotypes of the transformed state, and p53 has been identified as an immortalizing oncogene. Thus it is possible that p53-fixation by T antigen is responsible for its transforming potential. In this article, the transforming activities of T antigen and p53 are reviewed, and the possible relevance of p53 binding to T antigen-induced transformation is discussed. PMID- 3026532 TI - Forskolin stimulates cAMP production and the onset of the functional differentiation in the fetal rat thyroid in vitro. AB - Forskolin, a diterpen stimulating the adenylcyclase induces in vitro an accumulation of cAMP in thyroid glands explanted from 17 day-old rat fetuses. Thyroid glands from 15 day-old fetuses exposed to forskolin exhibit, within 24 hr, an active folliculogenesis and 125I fixation in follicular cavities. The results indicate that cAMP probably activates the onset of both morphological and physiological maturation in the fetal rat thyroid. PMID- 3026533 TI - Alkaline protease deficiency in the cr-1 (crisp) mutant of Neurospora crassa. AB - The adenyl cyclase deficient cr-1 mutant of Neurospora crassa grew poorly in bovine serum albumin as an alternative and only source of either sulfur, nitrogen or carbon. The low growth of the cr-1 mutant in protein was correlated with limited secretion of extracellular alkaline protease. The defect was specific for the cr-1 mutant and was suppressed by exogenous cyclic AMP. Cyclic AMP relieved protease deficiency under carbon, nitrogen or sulfur limiting conditions to unequal extents. Protease stimulation was greatest under carbon-limited conditions, but the resulting growth was least. Most of the cyclic AMP-mediated increase of alkaline protease was extracellular. PMID- 3026534 TI - Electrophoretic study of the genome of human rotavirus from Maceio, Brazil. AB - Rotaviruses were detected by enzyme immunoassay (EIA) in 53 (13.3%) of 397 fecal samples from children with acute gastroenteritis in the city of Maceio, Alagoas, Brazil. Polyacrylamide gel electrophoretic (PAGE) patterns characteristic of rotavirus double-stranded RNA were detected in 51 (96.2%) of the 53 EIA-positive samples. Of the RNA-positive samples, 1 (2%) was classified as subgroup 1 (short profile), 49 (96%) as subgroup 2 (long profile) and 1 (2%) could not be classified because of the absence of bands 10 and 11. The strains of subgroup 2 showed a great degree of electrophoretic heterogeneity and could be divided into several subcategories. Two samples showed splitting of one of the genome segments. PAGE, a very sensitive method capable of identifying rotavirus RNA genomes, has demonstrated that human rotaviruses detected in Maceio present many differences in RNA electrophoretic patterns. PMID- 3026535 TI - [An alternative method for the control of anti-aphthae vaccines]. PMID- 3026536 TI - Effects of nifedipine, verapamil, and diltiazem on [3H]-nitrendipine binding to the purified canine cardiac sarcolemmal membrane. PMID- 3026537 TI - Evidence for sympathetic, purinergic transmission in the mesenteric artery of the dog. AB - Electrical transmural stimulation of isolated mesenteric artery of the dog produced a transient contraction which consisted of adrenergic and non-adrenergic components. In contrast to the adrenergic component, the nonadrenergic component was resistant to prazosin and other adrenoceptor-blocking agents. However, the nonadrenergic component was completely blocked by guanethidine and by desensitization with alpha,beteta-methylene-ATP (alpha,beta-MeATP). Desensitization induced by alpha,beta-MeATP also inhibited the contractile response to ATP but not the adrenergic responses induced by electrical transmural stimulation and exogenous noradrenaline. These results suggest that the nonadrenergic contraction induced by electrical transmural stimulation is a sympathetic, purinergic response. PMID- 3026538 TI - Gastrointestinal neurotensin receptors: contribution of the aromatic hydroxyl group in position 11 to peptide potency. AB - Neurotensin structural analogues on tyrosine11 were tested in vitro to determine their ability to contract the fundus or relax the intestine. The rank order of potency was: neurotensin greater than [Phe11]-neurotensin greater than [D-Tyr11] neurotensin greater than [D-Phe11]-neurotensin. All peptides behaved as full agonists. It is concluded that tyrosine11 is part of the neurotensin pharmacophore; the hydroxyl group increases the affinity not the intrinsic activity of the peptide at the receptor. PMID- 3026539 TI - The effect of enalapril (MK421), an angiotensin converting enzyme inhibitor, on the conscious pregnant ewe and her foetus. AB - The effects of enalapril, an angiotensin converting enzyme (ACE) inhibitor, on maternal and foetal blood pressure, heart rate and components of the renin angiotensin-aldosterone system were studied in 9 chronically-cannulated pregnant ewes and their foetuses. Six ewes received 1 mg kg-1 enalapril i.v. while 3 were given 2 mg kg-1. Although the initial fall in blood pressure was slightly greater in the higher dose group, there was substantial overlap of data. The pressor response to angiotensin I, assessing ACE activity, was abolished within 10 min of administration, and did not recover during 3 h of observation. Maternal systolic and diastolic pressures reached a nadir 90 min after administration (P less than 0.001, P less than 0.002 respectively). The maximum tachycardia was seen at 60 min (P less than 0.05). The foetuses of the ewes given 1 mg kg-1 enalapril showed no change in systolic or diastolic blood pressure or heart rate. Those of the ewes given the higher dose showed late-onset hypotension, coincident with the lowest maternal blood pressures. Maternal plasma renin concentration (PRC) had risen significantly by 30 min (P less than 0.02), reaching a maximum at approximately 90 min. Maternal plasma angiotensin II and aldosterone concentrations both fell initially (P less than 0.05) but were almost at basal levels by the end of the experiment. Foetal plasma renin, angiotensin II and aldosterone concentrations were unchanged throughout the experiment. Peak values of enaprilic acid, the active principle, were recorded in maternal plasma 65-90 min after administration of 1 mg kg-1, and 25-30 min after the administration of 2 mg kg-1. A trace amount of the active principle was recorded in the foetal plasma of one lamb, whose mother had been given the higher dose. None was recorded in the plasma from three other lambs. Maternal plasma ACE concentrations fell by an average of 84%; in 4 of the 6 ewes in which concentrations were measured they were undetectable after treatment. Foetal plasma ACE concentrations were unchanged throughout. Enalapril at 1 mg kg-1 thus exerts a depressor effect on the pregnant ewe which is probably related to its blockade of the renin angiotensin system. Both direct measurement of the drug and foetal observation suggest that it does not cross the sheep placenta at this dose. This is consistent with its failure to cross the blood-brain barrier. Foetal effects were noted at 2 mg kg-1, and there was an unexpected foetal death in this group. PMID- 3026540 TI - Prostaglandin endoperoxide analogues which are both thromboxane receptor antagonists and prostacyclin mimetics. AB - Two prostaglandin endoperoxide analogues, EP 035 and EP 157, behave as specific thromboxane receptor antagonists on isolated smooth muscle preparations such as rabbit aorta, dog saphenous vein and guinea-pig trachea. However, in human platelet-rich plasma (PRP) they produce an unsurmountable block of aggregation induced by a wide range of agents (ADP, platelet-activating factor, thrombin); this inhibitory profile is typical of that seen with either prostaglandin I2 (PGI2) or PGD2. EP 035 and EP 157 induce large increases in cyclic AMP levels (up to 20 times basal) in human PRP. Simultaneous exposure to PGE1 markedly reduces their effect on cyclic AMP; exposure to PGD2 is much less effective in this respect. The adenylate cyclase inhibitor SQ 22,536 opposes the inhibitory action of EP 035, EP 157, iloprost (a stable PGI2 analogue) and PGD2 on platelet aggregation. However, the xanthone derivative AH 6809 blocks the inhibitory action of PGD2 but does not affect EP 035, EP 157 and PGI2 and its structural analogues. EP 035 and EP 157 displace [3H]-iloprost binding to the PGI2 receptor on human platelet membranes. Displacing ability is ranked as follows: iloprost greater than 6a-carba PGI2 greater than EP 157 greater than EP 035 greater than EP 164 (alpha-dinor derivative of EP 157). This order of potency is the same as that found for activation of adenylate cyclase in homogenates of washed human platelets and for inhibition of aggregation in washed human platelets. The activities of EP 035 and EP 157 were studied in two other systems containing PGI2 receptor-adenylate cyclase complexes, the NCB-20 cell line and human lung tissue. In both cases stimulation of adenylate cyclase was found but maximum rates were below that achieved with iloprost. These effects of EP 035 and EP 157 could be correlated with their abilities to displace [3H]-iloprost binding. 5 These results indicate that EP 035 and EP 157 inhibit the aggregation of human platelets by acting as agonists at the PGI2 receptor linked to adenylate cyclase. They represent a class of compound with both thromboxane receptor blocking activity and prostacyclin mimetic activity. PMID- 3026541 TI - Autoradiographic analysis of the distribution of beta-adrenoceptors in the dog splenic vasculature. AB - The technique of in vitro labelling and autoradiography has been used to localize beta-adrenoceptors in sections of the splenic vascular bundle of the dog. Binding of (-)-[125I]-cyanopindolol (Cyp) to sections of splenic vascular bundle equilibrated within 150 min and slowly dissociated after addition of (-) propranolol. The process was saturable with a dissociation constant (KD) of 40.3 +/- 4.4 pM and Bmax of 18.9 +/- 1.7 fM (in 6 sections). Binding to sections was stereoselective, the (-)-isomer of propranolol being 90 times more effective than the (+)-isomer in competing for (-)-[125I]-Cyp binding. Delineation of beta adrenoceptor subtypes using the selective antagonists betaxolol (beta 1) and ICI 118,551 (beta 2) indicated that the receptors present were almost exclusively of the beta 2-subtype. Autoradiographic studies under the conditions evaluated in the biochemical experiments showed that beta-adrenoceptors are unevenly distributed in the dog splenic vein, artery and associated nerve bundles. High concentrations of receptors are associated with the splenic nerves and lower but still significant concentrations in the vasculature. Higher resolution studies with nuclear emulsion coated coverslips revealed concentrations of beta adrenoceptors over cells adjacent to the lumen in veins. In arteries most beta adrenoceptors were found associated with the medial layer with fewer receptors towards the intima or adventitia. Serial sections of either artery or vein incubated with (-)-[125I]-Cyp in the presence of (-)-propranolol showed low levels of non-localized binding. PMID- 3026542 TI - Opioid inhibition of adenylate cyclase in the striatum and vas deferens of the rat. AB - The activity of adenylate cyclase in striatal membrane-enriched fractions (25,000 g) was inhibited by morphine, beta-endorphin, [D-Ala2-D-Leu5] enkephalin (DADLenk), fentanyl and bremazocine. Whereas guanosine triphosphate (GTP) appeared essential for the expression of this effect, sodium chloride seemed to enhance the degree of inhibition. Dopamine stimulation and sodium fluoride activation of the enzyme was also suppressed by morphine, beta-endorphin and DADLenk. beta-Endorphin and DADLenk inhibited adenylate cyclase activity in vasa deferentia membrane-enriched fractions (25,000 g); both opioids required GTP and NaCl and were inhibited by a delta-opioid receptor antagonist and by naloxone. Morphine, bremazocine and tifluadom did not significantly alter the activity of the vas deferens enzyme. Basal cyclic AMP values of striatal slices were not significantly altered by morphine, beta-endorphin or DADLenk. However, dopamine induced elevation of cyclic AMP was reduced by morphine and this effect of the opiate was suppressed by naloxone. Only beta-endorphin lowered the basal cyclic AMP values in the vas deferens. The physiological relevance of adenylate cyclase coupling to opioid receptor subtypes is considered. PMID- 3026543 TI - Vasoactive intestinal peptide in bovine pulmonary artery: localisation, function and receptor autoradiography. AB - The role of vasoactive intestinal peptide (VIP) in the control of pulmonary vascular tone was investigated by functional response, immunocytochemical localisation and receptor autoradiography in bovine pulmonary arteries. VIP immunoreactive nerve fibres were present at the adventitial-medial junction and in the media of the vessels. Exposure of precontracted bovine pulmonary artery segments to VIP in vitro resulted in almost complete (86 +/- 3%; mean +/- s.e.mean) relaxation, the concentration needed for 50% relaxation being 4.47 +/- 0.37 X 10(-9)M. VIP effects did not depend on the presence of intact endothelial cells. The distribution of VIP receptors was studied by autoradiography using [125I]-VIP. A high density of VIP receptors was found in arterial vascular smooth muscle, with a gradient of density from adventitia to luminal surface. There were no receptors on endothelial cells. These data show that VIP is a potent vasodilator of bovine pulmonary arteries, via direct activation of VIP receptors in vascular smooth muscle. VIP-immunoreactive nerves may influence pulmonary vascular tone directly and could, therefore, be important in regulating pulmonary blood flow. PMID- 3026544 TI - Bremazocine is an agonist at kappa-opioid receptors and an antagonist at mu opioid receptors in the guinea-pig myenteric plexus. AB - The agonist and antagonist activity of bremazocine at opioid receptors in the guinea-pig myenteric plexus preparation was determined in untreated tissues and in tissues in which either mu-9 or kappa-opioid receptors were blocked preferentially. After pretreatment of the tissue with beta-funaltrexamine for 90 min followed by washing out, the IC50 value of the selective mu-ligand [D Ala2,MePhe4,Gly-ol5]enkephalin was increased 67 fold whereas the IC50 values of the selective kappa-ligand U-69,593 and of the non-selective kappa-ligand bremazocine were not significantly changed. In this experimental design bremazocine acted only on kappa-receptors. After pretreatment of the tissue with beta-chlornaltrexamine and 10 microM of the mu-ligand for 30 min followed by washout, the IC50 value of the mu-ligand was increased 2 fold whereas the IC50 value of the selective kappa-ligand was increased 32 fold and that of bremazocine 62 fold. Under these experimental conditions, it was shown that bremazocine is an antagonist against [D-Ala2,MePhe4,Gly-ol5]enkephalin at the mu-receptor (Ke = 1.6 nM). The residual agonist activity of bremazocine is at the kappa-receptor. In naive myenteric plexus preparations the mu-antagonist activity of bremazocine cannot be demonstrated because its potency at the kappa-receptor is very high. This dual action may be of importance for the responses of bremazocine in other peripheral and central tissues. PMID- 3026545 TI - An investigation into the alpha-adrenoceptor mediating renal nerve-induced calcium reabsorption by the rat kidney. AB - An investigation was undertaken in pentobarbitone-anaesthetized rats to determine the sub-type of alpha-adrenoceptor responsible for the renal nerve-induced increases in the reabsorption of calcium and sodium by the tubules of the kidney. Stimulation of the renal nerves at low frequencies (0.8-1.5 Hz) did not change either renal blood flow or glomerular filtration rate but significantly reduced urine flow by 32%, calcium excretion by 36% and absolute and fractional sodium excretions by 36% and 22%, respectively. In the presence of the selective alpha 1 adrenoceptor antagonist prazosin, renal nerve stimulation (2-3 Hz) caused a significant reduction in renal blood flow of 7% but did not change either glomerular filtration rate, urine flow, calcium excretion or absolute and fractional sodium excretions. During administration of the selective alpha 1 adrenoceptor antagonist, idazoxan, renal nerve stimulation (1.0-1.5 Hz) significantly reduced renal blood flow by 4% and glomerular filtration rate by 7%; at the same time there were significant falls in urine flow of 43%, calcium excretion of 43% and absolute and fractional sodium excretions of 41% and 37%, respectively. These results show that low frequency renal nerve stimulation causes an anticalciuresis, independent of renal haemodynamics, which represents an increase in tubular reabsorption of calcium. This effect was blocked by prazosin but not idazoxan which is consistent with the mediation of alpha 1 adrenoceptors. The neurally-induced antinatriuresis also appeared to be dependent on the activation of alpha 1-adrenoceptors. PMID- 3026547 TI - Atriopeptin II-induced relaxation of rabbit aorta is potentiated by M&B 22,948 but not blocked by haemoglobin. AB - We examined the effects of haemoglobin (which inhibits the vascular responses to stimulation of soluble guanylate cyclase) and of M&B 22,948 (which selectively inhibits cyclic GMP phosphodiesterase) on the relaxation induced in rabbit aorta by the atrial natriuretic peptide, atriopeptin II (which stimulates particulate guanylate cyclase). Pretreatment with M&B 22,948 (100 microM) produced a 2.3 fold potentiation of atriopeptin II-induced relaxation of endothelium-denuded rings of rabbit aorta. Pretreatment with haemoglobin (10 microM) had no effect on the relaxation or the 10.9 fold increase in cyclic GMP content induced by atriopeptin II in endothelium-denuded rings of rabbit aorta. The potentiation by M&B 22,948 suggests a causal role for cyclic GMP in mediating atriopeptin II-induced vasodilatation of rabbit aorta. The inability of haemoglobin to block the atriopeptin II-induced rise in cyclic GMP suggests that it does not block stimulation of particulate guanylate cyclase. Thus, it is unlikely that a ferrous haem-containing receptor site is involved in the activation of the particulate form of guanylate cyclase as it is with soluble guanylate cyclase. PMID- 3026546 TI - Role of cyclic GMP in the modulation by endothelium of the adrenolytic action of prazosin in the rat isolated aorta. AB - The effect of endothelium on the adrenolytic action of prazosin was studied in the rat isolated aorta. Prazosin showed a non-competitive type of antagonism in preparations with intact endothelium while in preparations where endothelium had been removed, prazosin at concentrations between 0.3 nM-10 nM acted as a competitive antagonist. Methylene blue, used to decrease tissue levels of guanosine 3':5'-cyclic monophosphate (cyclic GMP), converted prazosin from a non competitive antagonist into an apparently competitive antagonist in the presence of endothelium. Increasing tissue levels of cyclic GMP by incubation with 8-bromo cyclic GMP converted prazosin from an apparently competitive antagonist into a non-competitive antagonist in the absence of endothelium. Analysis of concentration-response curves for noradrenaline in the presence and absence of endothelium showed that the affinity for noradrenaline was the same but the efficacy, measured by estimating the receptor reserve, was not; it was lower in the presence than in the absence of endothelium. It was concluded that the change in the mode of antagonism of prazosin after endothelium removal could be related to an alteration in the efficacy of the agonist, brought about by a change in the tissue levels of cyclic GMP. PMID- 3026549 TI - Herpes simplex virus encephalitis in children. PMID- 3026548 TI - Action of enflurane on cholinergic transmission in identified Aplysia neurones. AB - Effects of enflurane on the cholinergic transmission in Aplysia neurones were studied by current and voltage clamp methods. Acetylcholine (ACh) evoked three types of postsynaptic responses on different identified neurones: (1) a depolarizing response due to an increase in Na and K conductances (D-response), (2) a fast hyperpolarizing response due to an increase in C1 conductance (C1 response), and (3) a slow hyperpolarizing response due to an increase in K conductance (K-response). Enflurane altered neither the action potential nor the membrane resistance of the neurones but depressed the three ACh-induced responses, non-competitively, in a dose-dependent manner. The K-response was less suppressed than the other two. Blockade of the closed state of ion channel was suggested by a reduction in the first ACh response evoked 1 min after administration of enflurane. The anaesthetic facilitated the decay of the neurally evoked e.p.s.c. and i.p.s.c. in suggesting a reduction in the mean open time of the postsynaptic ion channel. It is concluded that enflurane depresses excitatory and inhibitory cholinergic transmission by reducing the postsynaptic currents. PMID- 3026550 TI - Carcinoma in situ of contralateral testis in patients with testicular germ cell cancer: study of 27 cases in 500 patients. AB - Carcinoma in situ in the contralateral testis was diagnosed in 27 of 500 patients (5.4%) with unilateral testicular germ cell cancer. Eight of the 27 patients received intensive chemotherapy for spread of their initial testicular cancer. Follow up biopsy studies did not detect changes of carcinoma in situ in any of these patients, and none developed a contralateral testicular tumour (observation time 12-88 months). Of the remaining 19 patients with carcinoma in situ, seven developed contralateral testicular cancer. The estimated risk of developing invasive growth was 40% within three years and 50% within five years. None of the 473 patients without carcinoma in situ detected by screening biopsy developed contralateral testicular cancer (observation time 12-96 months). No serious complications arose from the biopsy procedures. All patients with unilateral testicular germ cell cancer should be offered biopsy of the contralateral testis. PMID- 3026551 TI - The clinical features of HIV infection in Africa. PMID- 3026552 TI - Vegetarian diet in mild hypertension: a randomised controlled trial. AB - In a randomised crossover trial 58 subjects aged 30-64 with mild untreated hypertension were allocated either to a control group eating a typical omnivorous diet or to one of two groups eating an ovolactovegetarian diet for one of two six week periods. A fall in systolic blood pressure of the order of 5 mm Hg occurred during the vegetarian diet periods, with a corresponding rise on resuming a meat diet. The main nutrient changes with the vegetarian diet included an increase in the ratio of polyunsaturated to saturated fats and intake of fibre, calcium, and magnesium and a decrease in the intake of protein and vitamin B12. There were no consistent changes in urinary sodium or potassium excretion or body weight. In untreated subjects with mild hypertension, changing to a vegetarian diet may bring about a worthwhile fall in systolic blood pressure. PMID- 3026553 TI - Prevalence of antibody to poliovirus. PMID- 3026554 TI - Chronic demyelinating peripheral neuropathy associated with multifocal central nervous system demyelination. AB - A series of 6 cases is described in which a chronic demyelinating neuropathy was associated with a relapsing multifocal CNS disorder, the clinical features of which resembled multiple sclerosis. Multifocal CNS lesions were demonstrated by CT and MR imaging and the presence of CNS demyelination was indicated by prolonged central conduction times. These cases are discussed in relation to the occurrence of combined peripheral and central demyelination in chronic relapsing experimental allergic encephalomyelitis and neuritis. PMID- 3026555 TI - Phenytoin reduces early acetylcholine release after depolarization. AB - Phenytoin 10 microM inhibits the K-evoked release of acetylcholine (ACh) from synaptosomes, a process which is biphasic. Phenytoin acts only on the early phase of release. Replacement of external Na with Li does not modify phenytoin's effect. Phenytoin augments the spontaneous release of ACh from resting synaptosomes but this effect is eliminated in Li media. It is likely that phenytoin reduces K-evoked Ca uptake and the Na/Ca exchange by separate mechanisms. PMID- 3026556 TI - Characterization of glycine uptake in plasma membrane vesicles isolated from cultured glioblastoma cells. AB - C6 glioblastoma cells in culture were employed to isolate plasma membrane vesicles. After disruption of the glioblastoma cells by homogenization, membrane fractions were obtained by centrifugation on a discontinuous Ficoll density gradient. Fragmented membranes were found mainly in vesicular form. Transport of glycine has been demonstrated in membrane vesicles, using artificially imposed ion gradients as the sole energy source. The uptake of glycine is strictly dependent on the presence of Na+ and Cl- in the medium, and the process can be driven either by an Na+ gradient (out greater than in) or by a Cl- gradient (out greater than in) when the other essential ion is present. The process is stimulated by a membrane potential (negative inside) as demonstrated by the effect of ionophore valinomycin and anions with different permeabilities. The kinetic analysis shows that glycine is accumulated by two systems with different affinities. PMID- 3026557 TI - Increased food intake after opioid microinjections into nucleus accumbens and ventral tegmental area of rat. AB - The nucleus accumbens (ACC) and ventral tegmental area (VTA), two areas believed to subserve reinforcement and increases in locomotor activity produced by opioid microinjections, were examined for their involvement in opioid-produced changes in ingestive behavior. Opioids were infused bilaterally, and food and water intakes were measured for 1 h thereafter. Different morphine doses were administered and, with placements in globus pallidus and lateral ventricles as controls for diffusion, it was found that only ACC and VTA microinjections (0.1 10 nmol) produced dose-related increases in food intake. In both the ACC and VTA low doses of morphine also produced increases in water intake while in ACC high doses produced a decrease. Administration of morphine and an enkephalin analogue (Tyr-D-Met-Gly-Phe(4-NO2)-Pro-NH2) at different depths in the ACC indicated that the increase in food intake occurred at a site separate from that of the decrease in water intake. Using levorphanol, dextrorphan and morphine mixed with naloxone, it was shown that the effects were due to activation of opioid receptors. Additional experiments demonstrated that food intake is increased by ACC morphine under different levels of deprivation, with different times of testing and with availabilities of various goal objects in addition to food. The effect also did not appear to undergo development of tolerance or sensitization. It was concluded that there are sites in the ACC and VTA where increased activity of endogenous opioid peptide systems reliably increase food intake and it was hypothesized that these sites may contribute to changes in ingestive behavior after systemic morphine administration. Also, together with other effects produced by opioid microinjections into the ACC and VTA, the present findings suggest that increased opioid activity in these areas produce a pattern of behaviors similar to that produced in normal animals by food conditioned stimuli. PMID- 3026558 TI - Characterization of rat schwannoma-Schwann cell hybrids. AB - Sciatic nerve Schwann cells from strain LEC rats, homozygous for the c form of 6 phosphogluconate dehydrogenase (6-PGD), and RN22 rat Schwannoma cells, a subclone of RN2 deficient in hypoxanthine phosphoribosyltransferase and expressing the s form of 6-PGD, were fused to produce 'RNS' hybrid clones which proliferate rapidly in a medium containing hypoxanthine, aminopterin and thymidine (HAT) and express c, s and c/s heterodimeric forms of 6-PGD. RNS cells, like both parents, maintain a high baseline activity of 2', 3'-cyclic nucleotide 3'-phosphohydrolase and, as in RN22, activity of this enzyme is further inducible by 1 mM N6, O2' dibutyryl 3', 5'-cyclic AMP. The RNS clones resemble normal Schwann cells in the capacity to bind radioiodinated axolemmal fragments to their plasma membranes. PMID- 3026559 TI - Ro 15-1788 suppresses the development of kindling through the benzodiazepine receptor. AB - beta-Carboline (norharman) has been shown to produce kindled seizures when given systemically for long periods of time. The expression of the kindled seizure activity can be blocked by ligands of the benzodiazepine receptor suggesting the receptor as a site of vulnerability in the kindling mechanism. The present data show that Ro 15-1788, a receptor antagonist, suppresses the development of kindled seizures as demonstrated by the delayed appearance of each behavioral stage and the decreased severity of symptoms within each stage. Animals treated with Ro 15-1788 still expressed lower behavioral stages when Ro 15-1788 was eliminated from the trials indicating that this compound suppresses the very process of kindling itself and not just the expression of the kindled seizures. Ro 15-1788 given with norharman causes an increase in Bmax of [3H]flunitrazepam binding to the benzodiazepine receptor in cortex. It is not known if this increase is instrumental in lowering the kindling rate. PMID- 3026560 TI - Habituation of the local cyclic GMP response during amygdaloid carbachol kindling in the rat. AB - Seizures kindled with amygdaloid carbachol injections are transynaptic, dependent on activation of a specific population of muscarinic receptors, and some components of their expression could be mediated by intracellular second messengers. We measured cyclic GMP and cyclic AMP concentrations in micropunch biopsies of multiple brain regions after microwave fixation during the development and the expression of carbachol-kindled seizures in the rat. In the naive carbachol-injected amygdala, cyclic GMP concentrations rose from 1.03 +/- 0.15 pmol/mg protein to 2.21 +/- 0.46 after 2 min, and significant rises occurred in caudate, hypothalamus and contralateral amygdala. This response did not occur in implanted controls, after injection of mock cerebrospinal fluid, or when carbachol actions were blocked with atropine. The rise in cyclic GMP progressively disappeared upon repeated stimulation (injected amygdala on tenth stimulation: 0.72 +/- 0.23 pmol/mg protein). However, a late rise in both cyclic GMP and cyclic AMP concentrations occurred in many brain regions during convulsive seizures. These data suggest that during the development of kindling, changes in neuronal and synaptic excitability are associated with changes in intracellular second messengers. PMID- 3026561 TI - Electrical coupling and gap junctions between neurosecretory cells in an insect. AB - Intracellular injections of Lucifer yellow into single neurosecretory cells in the posterior (glandular) lobe of the corpus cardiacum of locusts resulted in dye transfer between these cells. Passage of hyperpolarizing or depolarizing current into one cell resulted in changes in the membrane potential of other cells. Freeze-fracture replicas of these cells revealed gap junctions which probably mediate the electrical and dye coupling. It seems apparent from these data that coupling may synchronize the neurosecretory cells' activity. PMID- 3026562 TI - Increased corticotropin-releasing factor receptors in rat cerebral cortex following chronic atropine treatment. AB - Rats were treated chronically with atropine (14 days, 20 mg/kg/day, s.c.) and corticotropin-releasing factor (CRF) receptors and CRF-mediated adenylate cyclase activity were measured in discrete brain regions. Chronic atropine treatment produced significant increases in muscarinic cholinergic receptors in the frontoparietal cortex (30% increase) and hippocampus (20% increase). No significant changes in the concentration of [125I]Tyr0-rat CRF binding sites were observed in olfactory bulb, cerebellum, striatum and hippocampus. In contrast, there was a significant and selective increase (35%) in CRF receptors in the frontoparietal cortex of atropine-treated rats. However, no significant corresponding changes in the Vmax or EC50 of CRF-stimulated adenylate cyclase activity accompanied the upregulation of CRF receptors in the cerebral cortex. These results demonstrate that CRF receptors in rat brain are subject to receptor regulation, the upregulation of CRF receptors occurs as a consequence of chronic muscarinic cholinergic receptor blockade, and this interaction between acetylcholine and CRF may be limited to the cerebral cortex. PMID- 3026563 TI - Regional distribution of high-affinity [3H]somatostatin binding sites in the human brain. AB - The distribution of high-affinity binding sites for [3H]somatostatin has been studied in membrane preparations from a number of regions of normal human brain. The highest densities of binding sites (greater than 48 fmol/mg protein) were found in the cerebral and cerebellar cortices and the hippocampus, with intermediate binding densities (30-46 fmol/mg protein) being present in the basal ganglia, amygdala, septum and claustrum. The lowest densities of binding sites (less than 14 fmol/mg protein) were observed in the hypothalamus, thalamus and substantia nigra. The binding of [3H]somatostatin in both the frontal cortex and cerebellar cortex demonstrated pharmacological specificity, since somatostatin 28, but not somatostatin-28(1-12) or Des AA1,2,4,5,12,13, D-Trp8-somatostatin, competed for the binding sites. Scatchard analysis of the binding in both frontal cortex and cerebellar cortex revealed the presence of two classes of high affinity binding sites. PMID- 3026565 TI - 2-Amino-7-phosphonoheptanoic acid, a selective N-methyl-D-aspartate antagonist, blocks swim-induced elevation of cerebellar cyclic guanosine monophosphate. AB - In order to explore how rapidly locomotor activity induces an elevation in cerebellar cyclic guanosine monophosphate (cGMP) content, Sprague-Dawley rats, pretrained to swim a 2.5-m course, were required to swim from one to 5 laps representing from 7 to 40s of strenuous activity. Immediately after completing the swimming task, each animal was killed by microwave irradiation and the cerebellum was collected for subsequent determination of the cGMP content. There was no difference in the cerebellar cGMP content between rats swimming one lap, i.e. for 7 s, and control rats that did not swim. However, there was a linear increase in the cGMP over control values from 1.8- to 2.4-fold in rats swimming 3 and 5 times, respectively. The first significant elevation of the cerebellar cGMP was seen at 24 s (3 laps). To determine if acidic amino acid pathways were involved in this elevation, a low dosage of a selective NMDA antagonist, 2-amino 7-phosphonoheptanoic acid (APH) was injected intracerebroventricularly 4 min before having rats swim 4 laps. This low dosage of APH, which alone had no effect on the cerebellar cGMP content, completely blocked the swim-induced elevation of this parameter. These data provide the first report of how quickly locomotor activity elevates the cerebellar cGMP content and further suggest that an NMDA receptor-mediated pathway is involved in the activity-induced elevation of this parameter. PMID- 3026564 TI - Differential depressive action of two mu and delta opioid ligands on neuronal responses to noxious stimuli in the thalamic ventrobasal complex of rat. AB - In the present investigation the effects of selective agonists for mu (Tyr-D-Ala Me-Phe-Gly-ol (DAGO)) and delta (Tyr-D-Thr-Gly-Phe-Leu-Thr (DTLET)) opioid receptors on neuronal activities induced by noxious cutaneous stimuli in the rat ventrobasal (VB) thalamus were analyzed. The two agonists produced a clear depressive action on thermal as well as mechanical noxious stimuli. The depressive action of DTLET (3 mg/kg i.v.) was lower and of shorter duration than that of DAGO (2 mg/kg i.v.). However, this effect is unambiguously related to the selective stimulation of opioid receptors since a consistent effect was also observed for a dose as low as 1.5 mg/kg i.v. of DTLET. Moreover, DTLET effect needs a high concentration of naloxone (0.5 mg/kg i.v.) to be reversed, while DAGO effect is totally reversed with 0.1 mg/kg i.v. PMID- 3026566 TI - Low concentrations of penicillin reveal rhythmic, synchronous synaptic potentials in hippocampal slice. AB - Field and intracellular recordings were used to examine the effects of varying concentrations of penicillin on synchronous CA3 activity in guinea pig hippocampal slices. In addition to the high-amplitude bursts, extracellular recordings in the distal apical dendrites (700-1200 microns from the soma) revealed biphasic mini field potentials (MFPs) which were not evident at the soma in 2000 IU/ml. A long-lasting (76 ms) field potential (A potential) with a waveform similar to the positive component of the MFP initiated the bursts. The cellular correlate of the positive component of the MFP and of the A potential appeared to be an EPSP and that of the negative component of the MFP and IPSP. Reductions of penicillin concentration below 2000 IU/ml (3.4 mM) decreased the burst rate and amplitude and increased burst threshold. At concentrations below 250 IU/ml the bursts were blocked and the MFPs increased in amplitude and occurred rhythmically at a mean frequency of 2.6 Hz. At intermediate concentrations the bursts arose from the rhythmic background. This activity more closely resembles that recorded with electroencephalography in human epileptic foci than does the high-dose penicillin preparation and may provide a better model of epileptiform discharge. PMID- 3026567 TI - Hippocampal sharp waves: their origin and significance. AB - This study investigated the spatial distribution and cellular-synaptic generation of hippocampal sharp waves (SPW) in the dorsal hippocampus of the awake rat. Depth analyses of SPWs were performed by stepping the recording electrode in 82.5 microns increments. SPWs were present during slow wave sleep, awake immobility, drinking, grooming and eating (0.01-2/s). The largest negative SPWs were recorded from the middle part of the stratum radiatum of CA1, the stratum lucidum of CA3, the inner molecular layer of the dentate gyrus and from layer I of the subiculum, in that order. The polarity of the SPWs was positive in layers II-IV of the subiculum, in stratum oriens and stratum pyramidale of CA1 and CA3, and in the hilus of the dentate gyrus. The electrical gradients across the null zones of the field SPWs were as large as 8-14 mV/mm. SPWs were associated with population bursts of pyramidal cells and increased discharges of interneurons and granule cells. During the SPW the excitability of granule cells and pyramidal cells to afferent volleys increased considerably. Picrotoxin and atropine and aspiration lesion of the fimbria-fornix increased either the amplitude or the frequency of SPWs. Diazepam and Nembutal could completely abolish SPWs. It is suggested that: hippocampal SPWs are triggered by a population burst of CA3 pyramidal cells as a result of temporary disinhibition from afferent control; and field SPWs represent summed extracellular PSPs of CA1 and subicular pyramidal cells, and dentate granular cells induced by the Schaffer collaterals and the associational fibers of hilar cells, respectively. The relevance of the physiological SPWs to epileptic interictal spikes and long-term potentiation is discussed. PMID- 3026568 TI - Effects of age on beta 1- and beta 2-adrenergic receptors in the brain assessed by quantitative autoradiography. AB - Aging female rats undergo a loss of reproductive cyclicity that has been correlated with alterations in norepinephrine neuronal activity in specific hypothalamic regions involved in the regulation of gonadotropin secretion. We explored the possibility that deteriorating reproductive function may also be due to changes in the density of receptors which bind and mediate responsiveness to norepinephrine. Existing evidence suggests that beta-adrenergic receptors mediate norepinephrine's inhibitory effect on luteinizing hormone (LH). Therefore, we determined whether changes in beta-adrenergic receptor densities occur in specific hypothalamic nuclei associated with reproductive function. Furthermore, we wished to determine whether age affects beta 1- and beta 2-adrenergic receptor densities differentially and whether age-related changes in receptor densities are related to hormonal status and/or vaginal smear cytology of the animals. Old constant estrous rats, old persistent diestrous rats, young cycling rats on estrus, and young cycling rats on diestrus were killed at 1000 h and brain beta 1 and beta 2-adrenergic receptor densities were measured using topical autoradiographic and computerized image analysis methods which permit a high level of neuroanatomical discrimination. We analyzed beta 1- and beta 2 adrenergic receptor densities in 7 brain areas including 4 hypothalamic regions known to be important in reproductive function. The pineal gland, the molecular layer of the cerebellum and the caudate putamen were examined because they contain high concentrations of beta-adrenergic receptors which have been reported to change with age. Serum luteinizing hormone was measured in the same rats. Serum LH concentrations were the same in all rats regardless of vaginal smear cytology and age.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3026569 TI - Protein kinase C in astrocytes: a determinant of cell morphology. AB - Protein kinase C-like activity was found to be present in astrocytes prepared from rat neocortex and maintained in culture. Exposure to phorbol 12-myristate 13 acetate (PMA) caused a redistribution of this kinase from the cytosol to the membrane fraction of these cells. Also PMA was found to cause a profound change in astrocyte morphology; cells were converted from flat, polygonal, undifferentiated cells to process-bearing cells. PMID- 3026570 TI - Anti-seizure and anti-epileptogenic effect of gamma-vinyl gamma-aminobutyric acid in amygdaloid kindling. AB - We examined the effects of systemic administration of gamma-vinyl gamma aminobutyric acid (GVG), a gamma-aminobutyric acid (GABA) transaminase inhibitor, on the kindling model of epilepsy in rats. GVG (1200 or 1500 mg/kg) approximately doubled the number of stimulations required for kindling development. GVG also suppressed both generalized motor seizures and electrographic after discharges in previously fully kindled animals. These results further support the idea that enhanced GABAergic neurotransmission suppresses both seizures and epileptogenesis. The results also suggest that GVG may be an effective anti seizure and anti-epileptogenic agent in humans. PMID- 3026571 TI - Motor actions of excitatory amino acids and their antagonists within the rat ventromedial thalamic nucleus. AB - The present study investigates the role of excitatory amino acid receptors within the rat ventromedial thalamic nucleus (VM) for the mediation of motor behaviour. For this purpose changes of motility were monitored after microinjections of excitatory amino acids and of various excitatory amino acid antagonists into the VM. N-Methyl-D-aspartate (NMDA) and kainate (KA), but not quisqualate (QA), led to a dose-dependent increase of exploratory activity. A specific NMDA antagonist (-)-2-amino-7-phosphonoheptanoate (-)-AP7), preferential non-NMDA antagonists, 1 (p-chlorobenzoyl)-piperazine-2,3-dicarboxylate (pCB-PzDA) and gamma-D glutamylaminomethylsulphonate (GAMS), and a broad spectrum antagonist, kynurenate (KYN), induced catalepsy in a dose-dependent manner. The catalepsy induced by (-) AP7 was antagonized by NMDA, but not by KA, the pCB-PzDA-induced catalepsy was blocked by KA, but not by NMDA and the KYN-induced catalepsy was reversed by either NMDA or KA. These data point to a role of both NMDA and KA receptors within the VM for the regulation of motility. PMID- 3026572 TI - Opposite effects of ventral tegmental and periaqueductal gray morphine injections on lateral hypothalamic stimulation-induced feeding. AB - Eating was induced in sated animals by lateral hypothalamic electrical stimulation following central microinjections of morphine or saline. With stimulation intensity fixed at a moderate level, time to eat 3 food pellets of 45 mg decreased with increases in stimulation frequency, approaching an asymptote near 7 s at approximately 70 Hz. Ventral tegmental area morphine injections (0.8 8 nmol) reduced the minimum frequency required to produce eating of 3 pellets within 20 s and reduced the frequency at which asymptotic performance was produced; it did not substantially change the speed of eating at which performance approached asymptote. Periaqueductal gray morphine injections (1.6-16 nmol) had the opposite effect; they increased the stimulation frequency required to meet either the 20 s (maximum) or the 7 s (asymptotic) response criterion. Since the ventral tegmental injections did not alter the absolute level of asymptotic performance, the changes in eating threshold associated with these injections are assumed to reflect motivational influences of morphine. PMID- 3026573 TI - Dopaminergic substrates of cocaine-induced place conditioning. AB - Recently, Spyraki et al. (Brain Research, 253 (1982) 195-203) reported that cocaine-induced place preference conditioning was unaffected by blockade of central dopamine (DA) or norepinephrine function. In addition, systemic injections of the local anesthetic procaine produced place preference conditioning. The present study was undertaken to further evaluate the possible role of DA in cocaine-induced place conditioning. In Expt. 1, a partial replication of Spyraki et al., systemic cocaine (5.0 mg/kg, i.p.) produced significant place conditioning that was not disrupted with the DA antagonist pimozide (1.0 mg/kg, i.p.). In Expt. 2, cocaine was microinjected unilaterally into the lateral ventricles to eliminate peripheral local anesthesia. Cocaine (50.0 micrograms, i.c.v.) produced place conditioning and pretreatment with pimozide (1.0 mg/kg, i.p.) disrupted the effect. In Expt. 3, place conditioning was not observed when cocaine presentations (50.0 micrograms, i.c.v.) were paired with both compartments. The substrates of cocaine-induced place conditioning were further investigated in Expt 4: Procaine (250 micrograms, i.c.v.) did not produce place conditioning whereas the DA agonist bromocriptine (50.0 micrograms, i.c.v.) did. Results suggest the involvement of central DA in cocaine-induced place conditioning. PMID- 3026575 TI - Beta-carbolines characterized as benzodiazepine receptor agonists and inverse agonists produce bi-directional changes in palatable food consumption. AB - Drugs which bind to specific benzodiazepine recognition sites fall into three categories: agonists, antagonists, and inverse agonists. A set of biochemical parameters is available which distinguishes between the three. In addition, actions of the drugs result in physiological and behavioural effects which are distinguishable. beta-Carboline derivatives provide a group of compounds which show high affinity for the benzodiazepine sites, and which contains examples belonging to each of the three categories. Evidence is reviewed which shows that beta-carboline benzodiazepine receptor agonists (ZK 93423, ZK 91296) produce increases in the consumption of a palatable diet by non-deprived rats, that beta carboline inverse agonists (FG 7142, DMCM) produce an anorectic effect, and that the beta-carboline ZK 93426 acts as a benzodiazepine receptor antagonist. The results support the proposal of bi-directional control of feeding responses through the action of drugs at a common benzodiazepine receptor. Furthermore, benzodiazepine receptor inverse agonists provide a novel class of anorectic agents. Evidence is also reviewed which is suggestive of the modulation of food related reward by drug actions at benzodiazepine receptors. PMID- 3026574 TI - Pharmacological and anatomical evidence of selective mu, delta, and kappa opioid receptor binding in rat brain. AB - While the distribution of opioid receptors can be differentiated in the rat central nervous system, their precise localization has remained controversial, due, in part, to the previous lack of selective ligands and insensitive assaying conditions. The present study analyzed this issue further by examining the receptor selectivity of [3H]DAGO (Tyr-D-Ala-Gly-MePhe-Gly-ol), [3H]DPDPE (2-D penicillamine-5-D-penicillamine-enkephalin), [3H]DSLET (Tyr-D-Ser-Gly-Phe-Leu Thr) and [3H](-)bremazocine, and their suitability in autoradiographically labelling selective subpopulations of opioid receptors in rat brain. The results from saturation, competition, and autoradiographic experiments indicated that the three opioid receptor subtypes can be differentiated in the rat brain and that [3H]DAGO and [3H]DPDPE selectively labelled mu and delta binding sites, respectively. In contrast, [3H]DSLET was found to be relatively non-selective, and labelled both mu and delta sites. [3H]Bremazocine was similarly non-selective in the absence of mu and delta ligands and labelled all three opioid receptor subtypes. However, in the presence of 100 nM DAGO and DPDPE, concentrations sufficient to saturate the mu and delta sites, [3H]bremazocine did label kappa sites selectively. The high affinity [3H]bremazocine binding sites showed a unique distribution with relatively dense kappa labelling in the hypothalamus and median eminence, areas with extremely low mu and delta binding. These results point to the selectivity, under appropriate conditions, of [3H]DAGO, [3H]DPDPE and [3H]bremazocine and provide evidence for the differential distribution of mu, delta, and kappa opioid receptors in rat brain. PMID- 3026576 TI - Autoradiographic localization of beta-adrenergic receptors in the rabbit pituitary. AB - The localization and characterization of beta-adrenergic receptors in the rabbit pituitary have been studied using [125I]cyanopindolol as a specific ligand. Slide mounted frozen sections were used for the autoradiographic localization and characterization of beta-adrenergic receptors. The displacement curves obtained from optical density of radioautograms or counting of scraped off sections demonstrated that beta-adrenergic receptors were mostly of the beta 2-subtype and highly concentrated in the intermediate lobe. Low concentrations of beta 2 adrenergic receptors were also found to be uniformly distributed in the anterior and posterior lobes. These results suggest that epinephrine and/or norepinephrine might play a physiological role in the regulation of the secretion of not only the intermediate lobe but also the anterior and posterior lobes of the rabbit pituitary. PMID- 3026577 TI - Decreased GABAergic innervation of the pituitary intermediate lobe after rostral hypothalamic cuts. AB - The effects of hypothalamic cuts at various rostro-caudal levels on the GABAergic innervation of the neurointermediate lobe of the pituitary gland have been studied. The GABAergic innervation was visualized through glutamate-decarboxylase (GAD) immunocytochemistry. Caudal hypothalamic cuts which transected the pituitary stalk completely abolished the GAD immunoreactive plexus. Rostral cuts which separated about one-third of the median eminence and arcuate nucleus from the pituitary gland decreased the GAD-immunoreactive network in the intermediate lobe but did not affect the neural lobe significantly. Although the precise location of the cell bodies giving rise to the GABAergic innervation of the neurointermediate lobe remains unknown, our findings indicate that their projections are descending ones. They are severed by rostral hypothalamic cuts and show a rostrocaudal arrangement. It is likely that the GABAergic endings of the intermediate lobe originate in the rostral hypothalamus, probably in the rostral part of the arcuate nucleus and/or in the anterior periventricular area. The GABAergic fibers in the neural lobe have a more caudal origin than those innervating the intermediate lobe. PMID- 3026578 TI - GABA-ergic control of alpha-melanocyte-stimulating hormone (alpha-MSH) release by frog neurointermediate lobe in vitro. AB - Measurement of glutamate decarboxylase (GAD) activity in the intermediate lobe of the frog pituitary and brain showed that neurointermediate lobe extracts represented 12% of the GAD activity detected in the whole brain. No significant activity was measured in distal lobe extracts. Immunocytochemical studies revealed GAD-containing fibers among the parenchymal cells of the pars intermedia. The localization of GAD-like material in the intermediate lobe of the frog pituitary suggested a possible role of gamma-aminobutyric acid (GABA) in the regulation of melanotropic cell secretion. Administration of GABA (10(-6) to 10( 4) M), to perifused neurointermediate lobes caused a brief stimulation of alpha melanocyte stimulating hormone (alpha-MSH) release followed by an inhibition. Picrotoxin (10(-4) M), a Cl- channel blocker, abolished only the stimulatory effect of GABA (10(-4) M), whereas bicuculline (10(-4) M), a specific antagonist of GABAA receptors, totally inhibited the effects of GABA (both stimulatory and inhibitory phases). Bicuculline induced by itself a slight stimulation of alpha MSH release, suggesting that GABA-ergic nerve fibers present in the intermediate lobe are functionally active in vitro. The GABAA agonist muscimol (10(-7) to 10( 4) M) mimicked the biphasic effect of GABA on alpha-MSH release. Administration of baclofen, a specific GABAB agonist (10(-7) to 10(-4) M) induced a dose dependent inhibition of alpha-MSH secretion. In contrast to GABA or muscimol, baclofen did not cause any stimulatory effect whatever the dose. Taken together these result suggested that GABAA and GABAB receptors were present on frog melanotrophs.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3026579 TI - Frontal cortex stimulation evoked neostriatal potentials in rats: intracellular and extracellular analysis. AB - Evoked potentials, action potentials and intracellular events were recorded in the neostriatum of urethane anesthetized rats to electrical stimulation of frontal cortex white matter, motor cortex and pre-limbic cortex. Five major waves of the evoked potential were identified. Wave N1 (3.9 msec latency) was small, preceded cellular events and probably represents activation of corticostriate terminals. Wave P1 (10.8 msec latency to peak following white matter stimulation) coincided with an EPSP and neuronal firing. Both wave N2 (38.0 msec latency to peak) and P2 (approximately 110 msec duration) overlapped the intracellularly recorded hyperpolarization and inhibition of cell firing. Based upon this correspondence and upon the behavior of waves N2 and P2 with changing current and during conditioning-test paired pulse stimulation, it was concluded that the waves represent different processes contributing to the cellular hyperpolarization. A late wave, N3 (175 msec onset latency) corresponded to a late rebound firing and cellular depolarization. This late wave was eliminated from the neostriatum, but not from the overlying sensorimotor cortex, by kainic acid lesions that destroyed medial thalamus but left thalamic lateral nuclei and reticular nucleus intact. PMID- 3026580 TI - Oscillatory behavior in inferior olive neurons: mechanism, modulation, cell aggregates. AB - Inferior olive neurons, in brain slices maintained in vitro, display spontaneous, continuous oscillations of their membrane potential which are consonant with olivary rhythmic activity seen in vivo. This oscillatory behavior was studied with intracellular electrophysiological techniques. The 3-10 Hz rhythmicity of these cells from guinea pigs is tetrodotoxin resistant and dependent on a somatic calcium conductance. The oscillatory behavior can exhibit intrinsic frequency modulation and can be altered by synaptic processes. Synaptic alteration of the oscillatory behavior by afferent sources and extensive electrotonic coupling between cells in local aggregates (shown by Lucifer yellow dye-coupling) provide the substrate for a potent central pattern generator with a well established efferent pathway for control of motor functions. PMID- 3026582 TI - Impact of food deprivation on alpha 1- and alpha 2-noradrenergic receptors in the paraventricular nucleus and other hypothalamic areas. AB - Hypothalamic norepinephrine may modulate normal eating behavior through activation of alpha 2-noradrenergic receptors, localized in the paraventricular nucleus (PVN). We investigated whether these receptors, which stimulate food ingestion, may in turn be altered by the nutritional state of the organism. Thus the impact of food deprivation, on the specific binding of [3H]-p-aminoclonidine ([3H]PAC) to alpha 2-noradrenergic receptors in discrete hypothalamic areas, was examined in rats. The results of our first experiment revealed that 48 hr food deprivation reduced (by 50%) the maximum number of binding sites (Bmax) of the high affinity component of [3H]PAC binding to alpha 2 receptors. This effect occurred exclusively in the medial hypothalamus (which includes the PVN), without any change in the affinity (Kd) of these receptors. A smaller decline was seen in the low affinity binding sites of the medial hypothalamus, whereas no changes were observed in the density or affinity of the high and low affinity alpha 2 receptor sites in the lateral hypothalamus or frontal cortex. The alpha 1 noradrenergic receptor sites, as defined by [3H]prazosin and [3H]WB-4101 binding, were also unaffected in the different brain areas by 48 hr food deprivation. An additional analysis of alpha 2 receptors in discrete hypothalamic nuclei demonstrated that the deprivation-induced decline in alpha 2-receptor binding: occurred specifically in the PVN; was apparent after as little as 3 hr food deprivation; and occurred only when this brief deprivation fell at the onset of the dark cycle, as opposed to at the end of the dark cycle.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3026583 TI - [Ambiguities of AIDS]. PMID- 3026581 TI - Raphe dorsalis-spinal cord cografts in oculo: electrophysiological evidence for an excitatory serotonergic innervation of transplanted spinal neurons. AB - Intraocular replicas of descending serotonergic bulbospinal pathways were constructed by means of sequential intraocular grafting of nucleus raphe dorsalis and spinal cord. Using extracellular recordings we have studied the functional connections between such double grafts. Superfusion of single spinal cord grafts with serotonin causes an increase in spontaneous activity. This excitation is reversibly blocked by the specific 5-hydroxytryptamine (5-HT) antagonist metergoline. Stimulation of the raphe part of nucleus raphe dorsalis-spinal cord double grafts causes a long-lasting excitation of the spinal neurons similar to that seen in single spinal cord grafts given serotonin. The electrically induced excitation could also be reversibly blocked with metergoline. It is concluded that serotonin-containing nerves from grafts of nucleus raphe dorsalis are not only morphologically organotypic, but also form functional contacts with neurons in cografted spinal cord. The results further support an excitatory or modulatory role of the descending spinal serotonergic pathways and demonstrate that functional contacts can be established between isolated CNS grafts when 5-HT fibers invade immature or mature spinal cord tissue. PMID- 3026584 TI - [Congenital soft tissue dysplasia: a new anatomicoclinical entity]. PMID- 3026585 TI - [Respective roles of aflatoxin and hepatitis B in the etiology of primary cancer of the liver in man]. PMID- 3026586 TI - [Melatonin and the corticotropic function]. PMID- 3026587 TI - Oncogene proteins and the insulin receptor. PMID- 3026588 TI - Transferrin receptor expression and the control of cell growth. PMID- 3026589 TI - [Development of gastric sensitivity to histamine in the rat]. AB - Sensitivity of the developing rat stomach to histamine (HA) was examined on isolated gastric mucosae of rats of various ages from the fetal to adult periods. Spontaneous acid secretion in mu eq/h.cm2 occurred at all the ages studied, at a basal rate of 0.45 +/- 0.07 in fetuses to 0.22 +/- 0.03 (day 5), 0.11 +/- 0.04 (day 10), 0.12 +/- 0.04 (day 12), 0.22 +/- 0.08 (day 16) and 0.33 +/- 0.04 (adults). In the fetal rats as in the adults, marked responses to respectively 10(-5) and 10(-4) M HA were demonstrated. The H2-receptor antagonist cimetidine diminished HA-induced secretion by 66 and 57% in fetuses and adults respectively. Between these two stages (from days 5 to 12), basal secretion and the response to HA dropped significantly. On day 21 of gestation, as well as on the critical days 5 and 12 after parturition, db-cAMP (10(-4) M) caused maximal stimulation of acid secretion. These results indicate that the development of responsiveness to HA in the rat is biphasic. They suggest that after birth, the H2-receptor adenylate cyclase system undergoes major modifications which might lead to the complete lack of responsiveness to HA by day 12. PMID- 3026590 TI - [Selective stimulation of histamine H3 autoreceptors]. AB - A simple derivative of histamine, alpha-methylhistamine i.e. 4-(2-aminopropyl) imidazole, was shown to potently inhibit the K+-induced release of [3H] histamine from slices of rat cerebral cortex previously incubated in the presence of [3H] histidine. The maximal inhibition elicited by alpha-methylhistamine was of about 60% i.e. similar to that elicited by exogenous histamine. The effect occurred with an EC50 value of 4.3 +/- 1.1 X 10(-9) M about 10 times lower than that of histamine and was reversed by a H3-receptor antagonist. Since alpha methylhistamine is known to display negligible potency at H1- and H2-receptors, this compound appears to be the first highly potent and selective H3-receptor agonist to be identified. PMID- 3026591 TI - Disaggregation of bone into crystals. AB - The sizes, shapes, and organizational states of the crystals in bone are studied by systematic disaggregation of the mineral phase. This is achieved by oxidizing the organic phase with sodium hypochlorite, dispersing the resultant particles by sonication, and separating the crystal aggregates from the crystal monomers by gravity setting in ethanol. Six different bones are compared. Bones in which crystals are intimately associated with the collagen fibrils mostly disaggregate into crystal monomers. In dense bones, where the crystals are mostly located between fibrils, they tend to persist as "fused" aggregates. All the crystals are tabular or plate-shaped. In bones in which the majority of crystals are associated with the collagen fibrils, just less than 90% of the crystals are shorter than about 450 A in length. Their widths are on the average about 250 A, almost an order of magnitude larger than the diameters of individual gap regions within the collagen fibril. The notion that one crystal is located in one gap region is therefore untenable and a reevaluation of the relations between collagen and mineral in bone is necessary. PMID- 3026592 TI - Gallium increases bone calcium and crystallite perfection of hydroxyapatite. AB - Gallium, a group IIIa metal, is known to interact with hydroxyapatite as well as the cellular components of bone. In recent studies we have found gallium to be a potent inhibitor of bone resorption that is clinically effective in controlling cancer-related hypercalcemia as well as the accelerated bone resorption associated with bone metastases. To begin to elucidate gallium's mechanism of action we have examined its effects on bone mineral properties. After short-term (14 days) administration to rats, gallium nitrate produced measurable changes in bone mineral properties. Using atomic absorption spectroscopy, low levels of gallium were noted to preferentially accumulate in regions of active bone formation, 0.54 +/- .07 microgram/mg bone in the metaphyses versus 0.21 +/- .03 microgram/mg bone in the diaphyses, P less than 0.001. The bones of treated animals had increased calcium content measured spectrophotometrically. Rats injected with radiolabeled calcium during gallium treatment had greater 45 calcium content compared to control animals. By wide-angle X-ray analyses, larger and/or more perfect hydroxyapatite was observed. The combined effects of gallium on bone cell function and bone mineral may explain its clinical efficacy in blocking accelerated bone resorption. PMID- 3026595 TI - Effect of soman (pinacolyl methylphosphonofluoridate) on the blood levels of corticosterone and adrenocorticotropin in mice. AB - Mice were poisoned by an extremely toxic organophosphate anticholinesterase soman (pinacolyl methylphosphonofluoridate), 50 or 100 micrograms/kg at 1000, and the serum concentrations of corticosterone were determined fluorometrically at 3-h intervals for at least 24 h. The lower soman dose (50 micrograms/kg) produced a modest increase in serum corticosterone concentrations but by 24 h the levels were not significantly different from control. Following the higher soman dose (100 micrograms/kg) the serum corticosterone levels were elevated significantly (p less than 0.05), for at least 27 h. However, ACTH concentrations were not elevated. It is possible that the elevated levels of corticosterone were due to a reduced metabolism and excretion of corticosterone resulting from the intense hypothermia, following soman poisoning which may change cardiac output and organ (liver and kidney) perfusion and not due to an enhanced release from the adrenal gland. PMID- 3026593 TI - Vitamin D3 and avian bone in vitro: specificity of effect on Japanese quail calvaria. AB - Calcitriol (1,25(OH)2D3) has been shown, under certain conditions, to elicit an in vitro response in adult avian calvarium which may be interpreted as calcium uptake by the bone. The present investigation was undertaken to study the specificity of this response. Calvaria were removed from 6-week-old female Japanese quail and cultured for periods of up to 96 hours at 37 degrees C in 5% CO2/95% air. 1,25(OH)2D3 induced a fall in the medium total and ionic calcium concentrations at both 48 hours and 96 hours of incubation; these responses were not blocked by the presence of 10(-4) M acetazolamide. Bovine parathyroid hormone (bPTH(1-34] at 10(-7) M, and dibutyryl cyclic AMP (DBcAMP) at 10(-4) M, had no effect on the medium calcium. In contrast, forskolin at 10(-4) M induced a marked fall in medium calcium concentrations, particularly at 48 hours. The specificity was also studied with respect to vitamin D3 and its two major metabolites. 1,25(OH)2D3 exhibited a bell-shaped dose-response relationship with the maximal effect at 10(-7) M. In contrast, the other two compounds elicited no effects at 10(-7) M or 10(-6) M; significant responses were observed at 10(-5) M with both agents. In general, 25-dihydroxyvitamin D3 (25OHD3) was more potent than vitamin D3. These findings suggest that the medium calcium response to 1,25(OH)2D3, interpreted as calcium uptake by the cultured adult avian bone, is relatively specific among calcemic agents; the response was elicited by forskolin but not by bPTH(1-34) or DBcAMP.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3026597 TI - Activity of pulmonary sarcoidosis. PMID- 3026596 TI - Sarcoidosis: past, present, and future concepts. PMID- 3026594 TI - Sarcolemmal Ca2+-binding and enzyme activities in myocardium from hypothyroid rat. AB - Effects of hypothyroidism on heart sarcolemmal activities were examined by using membrane preparations obtained by two different methods from rats treated with propylthiouracil for 6 to 8 weeks. ATP-independent Ca2+ binding, sialic acid and phospholipid content, Ca2+ ATPase, Mg2+ ATPase and adenylate-cyclase were not altered in membranes isolated by the hypotonic shock-LiBr treatment method from hypothyroid hearts. On the other hand, depressed activities of ouabain sensitive Na+-K+ ATPase and 5'-nucleotidase were observed in this hypothyroid preparation. Sarcolemma isolated by the sucrose density gradient procedure from hypothyroid hearts exhibited lower ouabain-sensitive Na+-K+ ATPase and higher ATP-dependent Ca2+ binding as well as Ca2+ stimulated ATPase without any changes in the 5' nucleotidase, adenylate cyclase and Mg2+-ATPase activities. The activation of ATP dependent Ca2+ binding and Ca2+ stimulated ATPase by calmodulin in the hypothyroid preparation was greater than the control; these effects of calmodulin were blocked by trifluoperazine. The results suggest some specific changes in the heart sarcolemmal Ca2+-pump during the development of hypothyroidism. PMID- 3026598 TI - Diagnosis of sarcoidosis. PMID- 3026599 TI - Granuloma formation and fibrosis in sarcoidosis. PMID- 3026600 TI - Relationship of cutaneous sarcoidosis to systemic disease. PMID- 3026601 TI - Cutaneous sarcoidosis: prognosis and treatment. PMID- 3026602 TI - Cushing's disease in a young woman with anorexia nervosa: pathophysiological implications. AB - This report describes a 17-year old student who was found to have Cushing's syndrome two years after she had developed anorexia nervosa (AN). The Cushing's syndrome was treated with bilateral resection of enlarged, hyperplastic, non tumorous adrenal glands. The diagnosis was further confirmed four years later when, two to three years after new symptoms had appeared, an ACTH secreting pituitary adenoma (that is, Cushing's disease) was found on surgery. The possible mechanism for the development of Cushing's disease in a patient with prior anorexia nervosa, a sequence of events reported once previously, is discussed. It is suggested that increased hypothalamic-pituitary corticotroph stimulation in association with the anorexia nervosa, a now well-established endocrine phenomenon, activated an occult, inactive pituitary basophil adenoma in this patient, eventually resulting in autonomous pituitary overproduction of ACTH by the tumor. PMID- 3026603 TI - Elevation of CA 125 in patients with benign and malignant ascites. AB - The presence of CA 125, an ovarian cancer-associated antigen, was assessed in serum and ascites from patients with ovarian cancer (n = 47), hepatocellular carcinoma (n = 21), and liver cirrhosis (n = 40). Abnormal levels of serum CA 125 were observed in 49% of ovarian cancer patients, and in 89% of these same patients with ascites. In all cases of cirrhosis or hepatocellular carcinoma with ascites, CA 125 levels were above 35 U/ml. The specificity and sensitivity of CA 125 measurement for detecting ascites in chronic liver diseases were 73% and 100%, respectively. Furthermore, the CA 125 level was always higher in ascites than in serum. The authors conclude that CA 125 is a nonspecific marker of ascites. This result must be considered in the interpretation of CA 125 elevation in patients with ovarian cancer. Moreover, CA 125 may be of value in the diagnosis and monitoring of peritoneal diseases. PMID- 3026604 TI - Differential retention of rhodamine 123 by avian sarcoma virus-induced glioma and normal brain tissue of the rat in vivo. AB - The time course of uptake, retention and clearance of the cationic lipophilic dye, rhodamine 123 (Rh123), within the central nervous system was qualitatively evaluated in rats. Weanling rats were injected intracerebrally with avian sarcoma virus, which induced malignant gliomas in situ before injection of Rh123. Comparison was made of the amount of fluorescence of Rh123 within the normal cerebral cortex, myelinated tracts of the brain, meninges, choroid plexus, and neoplastic foci at 1, 4, 8, 12 and 24 hours after intravenous injection. Fluorescence microscopy was utilized to identify tissues containing the dye. Normal neuropil did not contain Rh123 at any of the time periods studied. Gliomas retained the dye at 1, 4, 8 and 12 hours, with increasing uniformity of distribution and decreasing intensity of fluorescence over this time period. Fluorescence was not detected at 24 hours within the neoplastic tissues, but was evident at all time periods studied within the choroid plexus. The specific retention of Rh123 by malignant glioma and by the choroid plexus in vivo suggests that Rh123 may be useful for photochemotherapeutic treatment of brain neoplasms and disorders of the choroid plexus. PMID- 3026606 TI - Pathologic findings on re-excision of the primary site in breast cancer patients considered for treatment by primary radiation therapy. AB - Gross excision of the tumor followed by radiotherapy is used for treatment of early breast cancer. Local recurrence after this form of treatment is uncommon except in patients with infiltrating ductal carcinoma whose excision specimens reveal an extensive intraductal component (EIC). It is unclear whether this observation is due to a qualitative difference in the response of tumors with EIC to radiation or to a quantitative difference in the tumor burden remaining after gross excision in patients with EIC. To address this question, the authors examined pathologic material in 71 patients with infiltrating ductal carcinoma treated with gross excision of the tumor and then selected for re-excision of the tumor site prior to radiotherapy because of the presence of either EIC in the primary excision specimen or microscopic tumor at or close to margins in the initial excision. Residual carcinoma was seen in 62% of all patients but was more frequent among the 25 patients with EIC than the 46 without (88% vs. 48%, P = 0.002). The nature of the residual tumor differed for patients with and without EIC in the primary excision specimen. Residual carcinoma in patients with EIC was often widespread and composed predominantly of intraductal carcinoma. In contrast, residual tumor in patients without EIC usually consisted of only scattered microscopic foci of infiltrating and/or intraductal carcinoma. The authors conclude that patients with EIC treated with gross excision of the tumor frequently have considerable residual intraductal carcinoma near the primary site. This finding may account for the increased risk of local recurrence observed in patients with EIC treated without re-excision of the tumor site before radiation therapy. PMID- 3026605 TI - Long-term effects of intravenous hyperalimentation administered during intensive chemotherapy for small cell bronchogenic carcinoma. AB - Sixty-five patients with small cell bronchogenic carcinoma received their first two of three courses of intensive induction chemotherapy with (30 patients) or without (35 patients) intravenous hyperalimentation (IVH). Patients predominantly had extensive disease (55%), Zubrod's performance status 0 to 2 (63%) and less than or equal to 6% pretreatment weight loss (68%). Both treatment arms were comparable by prognostic factors. The chemotherapy included six remission induction courses of ECHO chemotherapy (E: epipodophyllotoxin VP-16-213; C: cyclophosphamide; H: hydroxydaunorubicin; O: oncovin [vincristine]) followed by six courses of maintenance with PRIME (PR: procarbazine; I: ifosfamide; ME: methotrexate). Prophylactic brain irradiation was given to all patients. Patients with limited disease received chest irradiation at the completion of ECHO. Fifty of 52 (96%) evaluable patients responded with a complete (56%) or partial (40%) remission. The complete remission (CR) rate was higher in the control arm (66% versus 43%; P = 0.11). Response duration and survival of patients was similar for both treatment arms. Combined median survival duration for all patients with limited and extensive disease was 15.75 and 11.50 months, respectively. Combined median survival duration for CR patients with limited and extensive disease was 25 and 13 months, respectively. Administration of IVH did not ameliorate the hematologic, gastrointestinal and infectious morbidity of ECHO chemotherapy. The IVH was effective in preserving body weight and improving delayed hypersensitivity reaction to a battery of skin test antigens. Administration of intensive ECHO chemotherapy to patients with small cell bronchogenic carcinoma resulted in high response rates, acceptable toxicities and improved survival. Administration of IVH did not improve the short- and long-term results of chemotherapy, and did not ameliorate its morbidity. Nutritional support, however, was helpful in preventing patient's weight loss. PMID- 3026607 TI - Alteration in osteoblast cell number and cell activity in the presence of invading malignant fibrous histiocytoma. AB - Correct prediction of tumor behavior and interpretation of local factors in the tumor microenvironment rely in part upon accurate determination of tissue changes after tumor invasion. The authors examined local bone changes in primary malignant fibrous histiocytoma (MFH) of bone in a 59-year-old woman. Three noninvolved and three tumor-involved sites were evaluated by quantitative determinations of bone structural and dynamic features. Compared to noninvolved sites, tumor-involved bone was characterized by significantly increased osteoblast index (89.4 +/- 15.6 [mean +/- SEM] versus 7.3 +/- 6.0, P = 0.008), percent osteoid area (12.1 +/- 2.7 versus 1.2 +/- 0.5, P = 0.02), percent of trabecular surface covered by osteoid (70.0 +/- 6.0 versus 14.5 +/- 4.8, P = 0.002), and percent osteoid lined by osteoblasts (36.4 +/- 3.6 versus 3.7 +/- 3.0, P = 0.002). Bone 7.8 mm distant from invading tumor cells showed features characteristic of noninvolved sites, whereas bone completely surrounded by tumor showed markedly decreased osteoblast features. Osteoblast function also was affected by tumor; the amount of matrix laid down per day bore a significant positive correlation with the osteoblast index. These data indicate the following: distinctive bone morphologic changes occur in situ during invasion by MFH; changes affect aspects of bone formation but not resorption during invasion; both osteoblast number and osteoblast activity are significantly altered; and changes are local in nature and probably reflect the osteoblast response to local tumor factor(s) and are dependent upon the extent of tumor invasion. PMID- 3026608 TI - Cushing's syndrome due to ectopic proopiomelanocortin gene expression by islet cell carcinoma of the pancreas. AB - Expression of proopiomelanocortin (POMC) was studied in a male patient with Cushing's syndrome and ectopic production of ACTH by a pancreatic carcinoma. Plasma ACTH levels (greater than 200 pg/ml) were elevated, and elevated serum cortisol and urinary free cortisol were partially suppressed to 25% of basal levels by high-dose dexamethasone. Petrosal and jugular vein sampling did not yield a gradient of ACTH. Immunohistochemical staining of tumor tissue removed at pancreatectomy was positive for ACTH and beta endorphin, and negative for corticotropin-releasing factor (CRF). Tumor cells cultured in vitro secreted ACTH and beta-endorphin, which comigrated with their respective radiolabeled standards on gel chromatography. Hydrocortisone suppressed in vitro ACTH secretion and CRF (100 nM) stimulated ACTH by 50% during 72 hours of incubation. Agarose gel electrophoresis of poly-(A) mRNA extracts of tumor tissue followed by hybridization with 32P-cDNA for POMC revealed 2 distinct RNA species. The major RNA species (about 1.0 kb) was smaller than authentic pituitary POMC mRNA (about 1.1 kb); a larger precursor band also was visualized, suggesting either processing or degradation of tumor-POMC mRNA. Cytoplasmic dot blot hybridization of tumor mRNA with POMC cDNA yielded a positive signal with increasing amounts of RNA blotted. Immunohistochemistry and radioimmunoassay (RIA) of ACTH, in vitro regulation of ACTH secretion, and expression of POMC mRNA species by this tumor document expression of the human POMC gene by an islet carcinoma associated with Cushing's syndrome. PMID- 3026609 TI - Unique variants of hCG in sera of choriocarcinoma patients. AB - Human chorionic gonadotropin (hCG) of pregnancy can be separated into several variants by an isoelectric focusing (IEF) method. The hCG produced by choriocarcinoma consists of components found in pregnancy sera (pI 3.9-6.0) and, in addition, unique variants with more acidic isoelectric points ranging from 3.2 to 3.7. The sera of all five patients with advanced choriocarcinoma contained these unique acidic variants of hCG and the immunoreactive levels were significantly higher in these patients than in normal pregnant women or patients afflicted with hydatidiform mole or invasive mole. When the acidic variants of hCG were treated with neuraminidase they migrated to the alkaline region (pI 9 11) of the IEF column. In conclusion, sera of choriocarcinoma patients contain unique variants of hCG. The finding that these highly acidic components can be converted to molecules with an alkaline pI by neuraminidase digestion suggests that this high negative charge may be due to an increased sialic acid content. Patients possessing the acidic variants of hCG in their sera had poor survival rate. Determination of the variants may be useful as a prognosis marker and in evaluating the efficacy of therapy. PMID- 3026610 TI - Chromosome instability in cell lineages of amniocyte clones morphologically transformed by Simian virus 40. AB - Seven clonally derived human amniocyte cell lines from independent Simian virus 40 transformation events were analyzed for chromosomal aberrations in the precrisis stage. The distribution of chromosomal aberrations over the haploid complement was nonrandom. With increasing passage number the number of involved chromosomes increased, but the type of aberrations were the same: dicentric chromosomes and translocations due, respectively, to fusion of telomeres and breakage in the centromere regions. Interruptions to telomeres were also evident from extrachromosomal material terminally on the p arms, especially, of chromosomes #7 and #11. The cytogenetic similarities between the present results and those in transfection experiments are mentioned. All seven clones became highly heterogeneous over 10 to 20 passages (i.e., 30-60 population doublings) when, in general, transformed amniocytes enter crisis. The chromosomes which were nonrandomly involved in aberrations exhibited great diversity in types of rearrangements. Thus, for each clone multiple different rearrangements for specific chromosomes were observed in different cells. Most of these aberrations had a single step origin, which implies that breakage occurred repeatedly for the same chromosome in different cells, suggesting that the specific chromosome had become unstable in an earlier cell generation. Thus, we observed development of clonal chromosome instability, a phenomenon not reported for viral transformed cells. PMID- 3026611 TI - Breakpoint cluster region rearrangements in chronic myelogenous leukemia with a masked Philadelphia chromosome. AB - Cytogenetic and molecular analyses were performed in a case of chronic myelogenous leukemia. The cytogenetic study revealed that the leukemic cells of this patient contained a three-way translocation involving chromosomes #5, #9, and #22, resulting in a masked Philadelphia chromosome; the karyotype of the leukemic cells was 46,XY,t(5;22;9)(q31;q11;q34). Southern blot analysis of leukemic cell DNA was performed using a 1.1 kb HindIII-EcoRI breakpoint cluster region (bcr) probe. The results showed that BglII digested DNA showed two abnormal bcr fragments (i.e., 5.2 kb and 2.7 kb) in contrast to the results with DNA from two patients with a standard Ph chromosome [t(9;22)(q34;q11)] who showed one normal 5.0-kb bcr fragment in addition to altered fragments of about 4.3 kb or 4.0 kb. Bam HI digests of DNA from the leukemic cells of the patient with the masked Ph chromosome showed two bands (3.3 kb and 6.5 kb), whereas, DNA from the two patients with standard Ph translocations showed only a 3.3-kb bcr fragment. The results indicate that the molecular events in a masked Ph affect the bcr locus in a manner similar to that seen in standard Ph chromosomes. PMID- 3026612 TI - Cytogenetic toxicity of paraquat and streptonigrin in Fanconi's anemia. AB - We reported earlier that the frequency of chromosomal aberrations observed in Fanconi's anemia lymphocyte cultures depends on the oxygen tension during growth of the cultures, suggesting that "activated oxygen species" (superoxide, O2-; hydrogen peroxide, H2O2; hydoxyl radical, OH; and singlet oxygen, 1O2) or other reactive products generated during oxygen metabolism may be involved in the production of chromosomal damage in this syndrome. Paraquat and streptonigrin, agents that have been proposed as model compounds exerting cellular toxicity through overproduction of superoxide, were tested for their clastogenic potency in lymphocyte cultures from healthy controls and patients with Fanconi's anemia. Paraquat, at concentrations that severely affected mitotic activity (100-200 micrograms/ml), appeared to be a weak clastogen in human lymphocytes, whereas a clastogenic effect of streptonigrin was demonstrable already at a concentration as low as 0.005 microgram/ml. The results indicate that Fanconi's anemia lymphocytes fail to exhibit an increased sensitivity to the antimitotic and clastogenic effects of paraquat and streptonigrin. This suggests that intracellular superoxide is not critically involved in the generation of spontaneous chromosomal aberrations in Fanconi's anemia. We infer from these and previous data that singlet oxygen (1O2) may be a critical contributor to the chromosomal breakage in this disorder. PMID- 3026613 TI - Biological, pathological and clinical features of small cell lung cancer. AB - Small cell lung cancer, which is not uncommon, and is one of the most malignant and relatively well investigated solid tumors of adults, has been reviewed concerning its biology, pathology and clinical aspects. Although it is histologically very simple, its poorly differentiated epithelial cell characteristics are complicated by features of neuroendocrine cells, such as amine and peptide hormone production and specific enzyme activities, some of which have been found to be good monitoring markers during and after treatment. Because of the relative ease of establishing cell lines in vitro, cell characteristics have been studied in detail. This has led to subtyping of cell lines and may further lead to subtyping of histology. However, accumulation of further evidence has disclosed exceptions and unclassifiable cell lines. The same can be said about chromosomal abnormality. The reactivity of monoclonal antibodies and also oncogenes supports the prevalent concept discriminating small cell cancer from non-small cell cancer. However, concepts concerning histogenesis are still changing. Although it is one of the solid tumors most sensitive to radiation and chemotherapy, the response rate of the tumor to non-surgical treatment appears to have reached a plateau. In order to make a breakthrough in the treatment, strategies based on biological findings must be applied. PMID- 3026614 TI - Lipoprotein-associated oncornavirus-inactivating factor in the genus Mus: effects on murine leukemia viruses of laboratory and exotic mice. AB - High titers of oncornavirus-inactivating factor (OIF) were found previously in sera of laboratory mice. OIF is highly active against mouse xenotropic and polytropic envelope recombinant murine leukemia viruses (MuLVs) but not against ecotropic MuLVs. Of the 20 different mouse species or subspecies currently tested, that represent 4 subgenera, no OIF was found in the 3 subgenera more distant to the laboratory mouse. In the subgenus Mus, 7 of the 8 most distant species had no OIF, whereas all the ancestral species and subspecies of the laboratory mouse (Mus musculus musculus, Mus musculus domesticus), including a more distant member (Mus musculus cookii), had ample titers of OIF. A new separation technique was devised so that potential virus-neutralizing immunoglobulins could be separated by electrophoresis from OIF in small-volume serum samples. Active OIF was recovered from serum high-density lipoprotein, from very-low-density lipoprotein, as well as from chylomicron fractions. Murine sarcoma virus pseudotypes were made with several available exotic MuLV types. These pseudotype MuLVs were not susceptible to standard OIF preparations. The sera of exotic mice also had no factor analogous to OIF which would inactivate their own homologous or heterologous exotic MuLVs. It appears that, with one exception, OIF activity is limited to two subspecies of M. musculus and may be correlated in these subspecies with the presence of endogenous xenotropic MuLVs. PMID- 3026615 TI - Induction of class I major histocompatibility complex antigens in human teratocarcinoma cells by interferon without induction of differentiation, growth inhibition, or resistance to viral infection. AB - The behavior of human teratocarcinoma cells, and especially their stem cells (embryonal carcinoma cells), may provide insights into the properties of human early embryonic cells. We report here that human recombinant gamma-interferon (IFN-gamma) induced the expression of major histocompatibility complex Class I (HLA-A, B, C) antigens and beta 2-microglobulin in the two human embryonal carcinoma cell lines, 2102Ep cl.4D3 and NTERA-2 cl.D1, and in the yolk sac carcinoma cell line, 1411H; human recombinant IFN-alpha and IFN-beta were less effective inducers of these cell surface molecules. No induction was observed in the gestational choriocarcinoma cell line, JAR. Neither IFN-alpha, IFN-beta, nor IFN-gamma caused growth inhibition, expression of major histocompatibility complex Class II (HLA-DR) antigens, resistance to viral (vesicular stomatitis virus) infection, or expression of 2',5'-oligo(A)synthetase in any of the cells. Also, IFN-gamma neither induced differentiation of NTERA-2 cl.D1 cells, which are pluripotent human stem cells, nor influenced their differentiation induced by retinoic acid. However, developmental regulation of responsiveness to IFN was evident, since IFN-gamma induced higher levels of surface expression of HLA-A, B, C and beta 2-microglobulin in the retinoic acid-induced differentiated NTERA-2 cl.D1 cells than in the undifferentiated parental cells. Also, 2',5' oligo(A)synthetase was inducible in the NTERA-2 cl.D1 differentiated cells by IFN alpha and -beta, although not by IFN-gamma, and slight resistance to vesicular stomatitis virus infection was evident in aged cultures of differentiated cells exposed to IFN-alpha. The effect of recombinant mouse IFN-gamma on major histocompatibility complex expression by several murine teratocarcinoma cells was also examined: H-2 Class I (H-2Db), but not class II (I-Ab), antigens were induced in the parietal yolk sac carcinoma lines, PYS and F9Ac cl.9; in cultures of PCC3/A/1 containing both embryonal carcinoma (EC) and differentiated cells; and in cultures of the EC cells, PCC4azaR and PCC4AO, without evidence of differentiation. No induction was observed in the murine EC cell lines, F9 or FA (H-2Kk). Our results indicate that human EC cells, like murine EC cells, exhibit only a partial response to the interferons, and that the extent of this response is developmentally regulated. PMID- 3026617 TI - Selective stimulation of small cell lung cancer clonal growth by bombesin and gastrin-releasing peptide. AB - Human small cell lung cancers (SCLC) produce and secrete the regulatory peptide bombesin (BN) or its mammalian counterpart gastrin-releasing peptide (GRP). In addition, several SCLC tumor lines have been shown to express high affinity receptors for BN/GRP. On the basis of these findings, we investigated the effect of exogenously added BN and GRP on the soft agarose colony growth of a panel of human cell lines. In serum-free defined medium, colony formation of 9 of 10 SCLC cell lines was stimulated up to 150-fold by BN or GRP, with peak colony stimulation observed at 50 nM BN. In contrast, no stimulatory effect of BN was observed on nine non-SCLC cell lines. Although no stimulation of colony growth by BN was seen in serum-supplemented medium, addition of BN to the serum-free medium increased cloning efficiency to that achieved by serum in most of the SCLC cell lines. GRP 1-27, the active mammalian analogue of Bn, stimulated colony growth of SCLC cells similar to the manner of BN, while the physiologically inactive BN analogue, des-Leu (13)-Met (14)-BN, had no effect on colony growth. No correlation was observed in SCLC cell lines between the response of these cells to exogenous BN and the amount of cellular BN/GRP produced or the presence of BN receptors. These data suggest that BN/GRP may in some instances function as an autocrine growth factor for SCLC and indicate new ways for modulating SCLC growth in patients with this tumor. PMID- 3026616 TI - Enhanced therapeutic efficacy of cisplatin by combination with diethyldithiocarbamate and hyperthermia in a mouse model. AB - A spontaneously metastasizing solid tumor model derived by transplanting the TA3Ha murine mammary carcinoma into the s.c. tail tissue of mice was used to develop a treatment strategy for enhancing the therapeutic efficacy of cisplatin (CDDP). This strategy was based on the findings that diethyldithiocarbamate (DDTC) reduces the toxicity of CDDP, and that localized hyperthermia (HT) augments the antitumor efficacy of CDDP. DDTC (500 mg/kg) reduced the CDDP induced nephrotoxicity and gastrointestinal toxicity as well as increased the CDDP LD10 from 8 to 20 mg/kg in strain A mice. When CDDP and DDTC were used in multiple treatment schedules at 5-day intervals, DDTC protected the hosts but not the tumors against the toxicity of CDDP. HT administered locally to the tumor 1 h after the injection of CDDP (8 mg/kg) in 1 ml Hanks' balanced salt solution increased the antitumor effect but not the host toxicity. While administration of 8 mg/kg CDDP alone or with HT three times at 5-day intervals caused 100% host mortality, this dose of CDDP could be used with no mortality by combining it with DDTC. A combination of 8 mg/kg CDDP with DDTC (750 mg/kg) and HT (43 degree C for 60 min), administered three times at 5-day intervals, retarded the local tumor growth significantly compared to the untreated, CDDP plus DDTC plus HT control groups of mice. The frequency of lung metastasis in these groups on day 30 of tumor inoculation were 0, 90, 90, and 80%, respectively. The mean survival days of the mice treated with CDDP plus DDTC plus HT was 61 +/- 6 compared to 34 +/- 5 in the controls. The results presented here demonstrate that by combining CDDP with DDTC, high doses of CDDP can be safely administered. When localized HT is combined with high dose CDDP and DDTC, the tumor growth retardation and the host survival prolongation are significantly better than those obtained with the highest tolerable dose of CDDP alone or CDDP plus HT. PMID- 3026618 TI - Monoclonal antibody that distinguishes small-cell lung cancer from non-small-cell lung cancer. AB - To examine whether a monoclonal antibody, TFS-4, can distinguish small-cell lung cancer from non-small-cell lung cancers, an extensive survey of fresh lung tumors, cancers from other organs, and normal tissue specimens has been carried out. The antibody has been shown to react specifically with small-cell lung cancer (15 of 15) but not with squamous cell carcinoma (0 of 20), adenocarcinoma (0 of 20) of the lung, or large-cell lung cancer (0 of 2). It reacted neither with other malignancies, including colorectal cancer, gastric cancer, and malignant lymphoma, nor with such normal tissues as trachea, lung, liver, pancreas, colon, kidney, spleen, skin, striated muscle, bone marrow, or peripheral blood cells. Interestingly, the antibody cross-reacted with central nervous tissues. The antigenic determinant on small-cell lung cancer and that on human brain were both heat labile and trypsin sensitive, but resisted treatment with neuraminidase, suggesting that they represent similar peptides. TFS-4 may be of clinical use in the diagnosis of small-cell lung cancer, while the antigen may help investigate the nature and origin of small-cell lung cancer. PMID- 3026619 TI - Reduction of hepatic metastases in rabbits by administration of an oily anticancer agent into the portal vein. AB - We studied a prophylactic chemotherapy against hepatic metastases arising from the shedding of tumor cells into the portal circulation. The therapy was done with a lymphographic oily contrast medium, Lipiodol, and a high molecular weight anticancer agent named poly(styrene-maleic acid) copolymer conjugated neocarzinostatin (SMANCS), developed in our laboratory. SMANCS was dissolved in Lipiodol by sonication (SMANCS/Lipiodol, 1 mg of SMANCS in 1 ml of Lipiodol). Twelve rabbits were simply inoculated with the highly malignant carcinoma VX-2. Fifteen rabbits were given injections of SMANCS in glucose and Lipiodol into the portal vein and were subsequently inoculated with the tumor cells. Eighteen were given injections of SMANCS/Lipiodol and then the tumor cells. These rabbits were killed 12 days later. Thirteen were given injections of the tumor cells alone and were allowed to survive. Sixteen were given injections of SMANCS/Lipiodol and then with the tumor cells; they were allowed to survive. Rabbits given injections of SMANCS/Lipiodol before tumor inoculation had significantly fewer (P less than 0.001) metastases than those not treated or those given SMANCS in glucose and Lipiodol. Survival was significantly longer [P less than 0.005; 36.0 +/- 7.7 (SD) days] with SMANCS/Lipiodol before tumor inoculation than without treatment [23.5 +/- 3.0 days]. SMANCS/Lipiodol has a prolonged anticancer effect because it remains in the portal vein and allows sustained drug release from the oil (Lipiodol) to aqueous spaces. Hepatic metastases might be prevented by portal administration of the appropriate oily anticancer agent. PMID- 3026620 TI - Sixth Sapporo Cancer Seminar. PMID- 3026621 TI - In vitro toxicity and DNA cleaving capacity of benzisoquinolinedione (nafidimide; NSC 308847) in human leukemia. AB - Benzisoquinolinedione (nafidimide; NSC 308847) is an investigational drug currently in phase I clinical testing. We have studied the antileukemic activity in vitro, the cellular drug transport, and the molecular mechanism of action with DNA of this new compound. By agarose gel electrophoresis, we verified that nafidimide is an intercalating agent, through its alteration of the electrophoretic migration of DNA products produced by the relaxing action of DNA topoisomerase I. Concentrations of up to 100 microM of nafidimide did not produce topoisomerase I-mediated DNA cleavage. Nafidimide produced DNA single-strand breaks (SSB), double-strand breaks, and DNA-protein cross-links in human myeloid leukemia cells (measured with filter elution). The ratio of SSB/DNA-protein cross links was 1.32 +/- 0.36, a value similar to that produced by 4'-(9 acridinylamino)methanesulfon-m-anisidide (m-AMSA), suggesting that nafidimide, like m-AMSA, produced protein-associated DNA-strand breaks through a topoisomerase II-mediated reaction. The production of double-strand breaks by nafidimide also suggests the involvement of topoisomerase II in the drug-induced DNA cleavage. The cytotoxic activity of nafidimide was quantified in human myeloid leukemia cell lines differing by a factor of 70 in their cytotoxic sensitivity to m-AMSA. The m-AMSA-resistant line was less than 2-fold resistant to nafidimide. Cellular drug uptake was rapid and reached a steady state level in 30 min at 37 degrees C. At the end of exposure, drug egress was rapid, as was the disappearance of the DNA SSB. Rapid cellular uptake of nafidimide, with low retention at the end of exposure and rapid rejoining of DNA SSB suggest that prolonged cellular exposure may be necessary for optimal antitumor effect. In vitro cloning data suggest that nafidimide may be a therapeutic option for patients with leukemia resistant to m-AMSA. PMID- 3026622 TI - Human brain tumor-associated urinary high molecular weight transforming growth factor: a high molecular weight form of epidermal growth factor. AB - Urinary protein obtained from a patient with a highly malignant brain tumor (astrocytoma, grade IV) was adsorbed to trimethylsilyl controlled-pore glass beads and selectively eluted with acetonitrile to yield a high molecular weight (HMW) human transforming growth factor (hTGF). This HMW hTGF promoted clonogenic cell growth in soft agar and competed for membrane receptors with mouse epidermal growth factor. After surgical resection of the tumor, no HMW hTGF was found in urine. HMW hTGF generated a human EGF (hEGF) radioimmunoassay competitive binding curve similar to that of hEGF and parallel to that of a highly purified HMW form of hEGF previously reported to be present in trace concentrations in normal human urine. Both hEGF and HMW hEGF were clonogenic in soft agar, and their clonogenic activity as well as that of HMW hTGF was inhibited by anti-hEGF serum. Both HMW hTGF and HMW hEGF had 20 to 25% of the radioreceptor binding activity of hEGF. HMW hTGF purified from the pooled urine of several patients with malignant astrocytomas and HMW hEGF purified from normal control urine comigrated at Mr 33,000. Thus, HMW hTGF was indistinguishable from HMW hEGF in terms of apparent molecular size, epidermal growth factor receptor binding activity, epidermal growth factor immunoreactivity, and clonogenic activity. Urinary HMW hEGF/hTGF may be of tumor cell origin or may represent a response of normal host tissues to the tumor or its products. PMID- 3026623 TI - Defects of cyclic adenosine 3':5'-monophosphate-dependent protein kinases in initiated clones derived from BALB/c 3T3 mouse fibroblasts. AB - Two-stage carcinogenesis is involved in the transformation of mouse fibroblasts BALB/c 3T3 cells. In order to investigate the role of cyclic adenosine 3':5' monophosphate (cAMP)-dependent protein kinase at the stage of initiation, the following experiments were carried out: (a) two initiated clones (M14, M20) which exhibit 12-O-tetradecanoylphorbol-13-acetate-dependent growth in soft agar medium were isolated from cells treated with N-methyl-N'-nitro-N-nitrosoguanidine. The activity of cAMP-dependent protein kinase in M14 was reduced while that in M20 was similar to the level in parental cells. However, cAMP-binding activity to a regulatory subunit of cAMP-resistant clones were isolated from 4-nitroquinoline oxide- or ethyl methanesulfonate-treated cells. These clones have reduced activities in both cAMP-binding and cAMP-dependent protein kinase itself. Two of three cAMP-resistant clones were proved to be able to grow in soft agar medium only in the presence of 12-O-tetradecanoylphorbol-13-acetate. PMID- 3026624 TI - Buspirone in Parkinson's disease. AB - Buspirone, an anxiolytic unrelated to benzodiazepines that may act at the presynaptic dopamine receptor, was given to 11 patients with Parkinson's disease in an open label study. Seven patients completed the initial 10 week study achieving doses of 50-70 mg/day without any significant change in their clinical status. Six patients continued for an additional 3-11 weeks with increases in dose to 65-100 mg/day. Two of the three most severely affected patients had mild worsening of parkinsonian symptoms. Buspirone is ineffective in the treatment of Parkinson's disease, but at anxiolytic doses (less than 40 mg/day) does not adversely affect parkinsonian disability. PMID- 3026625 TI - Phase I and pharmacokinetic study of trans-N3P3Az2(NHMe)4. AB - trans-N3P3Az2(NHMe)4, an aziridinyl-substituted cyclophosphazene, was tested for its toxicity, pharmacokinetic behavior, and cytostatic activity in a phase I study in 30 patients. A total of 66 courses of a single iv bolus injection were given in five dose steps. Toxicity consisted of leukocytopenia and thrombocytopenia, dose limiting at 70 mg/m2, mild anemia, and some nausea. Leukocyte and platelet count nadirs fell between 2 and 3 weeks, with recovery at 6 weeks. A tendency for cumulative thrombocytopenia was noticed in three of 13 patients at risk. A three-phase plasma elimination model was applicable with t1/2 alpha of 9.9 minutes, t1/2 beta of 78.5 minutes, and t1/2 gamma of 435.5 minutes; renal drug excretion was substantial. Three partial remissions in 21 evaluable patients suggest some clinical activity for this drug. PMID- 3026627 TI - Phase II trial of carboplatin in patients with advanced germ cell tumors refractory to cisplatin. AB - A phase II trial of carboplatin was conducted in 22 patients with advanced germ cell tumors refractory to cisplatin-based chemotherapy. Twenty of 22 patients were evaluable for toxicity and response. Two partial responses and one minor response were seen. The major toxic effect was myelosuppression. Carboplatin has antitumor activity in patients with advanced germ cell tumors refractory to cisplatin, and phase II and III studies of combination chemotherapy are indicated. PMID- 3026626 TI - High-dose carmustine with autologous bone marrow transplantation for the adjuvant treatment of high-grade gliomas of the central nervous system. AB - The long-term survival of patients with high-grade gliomas of the CNS is poor despite the use of radiation therapy and chemotherapy. In an attempt to improve the survival of such patients, we administered adjuvant chemotherapy as high-dose carmustine with autologous bone marrow transplantation. Eighteen patients, 15 with glioblastoma multiforme and three with anaplastic astrocytoma, were treated shortly after completion of standard radiation therapy. One course of carmustine was administered at a total dose of 900-1050 mg/m2 iv over 3 days followed in 3 days by the reinfusion of the previously cryopreserved bone marrow. Acute toxicity was mild, but eight patients had severe pulmonary or CNS toxicity of which four episodes of pulmonary toxicity were fatal. For all patients treated, the median survival was 17.5 months from diagnosis and the estimated probability of surviving greater than 27 months was 22% (95% confidence limits of 14%-32%). Presently, there are four long-term survivors, two with progressive tumor, one suffering from severe encephalomyelopathy, and one alive and well. Although, in this series, we have observed long-term survivors, the overall estimated probability of survival is not substantially different from conventional treatment. In addition, the 22% incidence of fatal pulmonary toxicity suggests that this treatment will not measurably add to the treatment of CNS gliomas. PMID- 3026628 TI - Transient nephrogenic diabetes insipidus following high-dose cyclophosphamide chemotherapy and autologous bone marrow transplantation. PMID- 3026629 TI - Hypokalemia modulates alpha- and beta-adrenoceptor bindings in rat skeletal muscle. AB - Changes in the population of adrenergic alpha- and beta-receptors were examined in rat soleus muscles during hypokalemia by their direct determination using radiolabeled ligands. Only beta-adrenoceptors were detected in the normal rat muscles. Hypokalemia led to a pronounced decrease in beta-adrenoceptors, the number of [3H]DHA binding sites, by 50%, as compared with that in the normal rats. There was a genesis of alpha 1-adrenoceptors in hypokalemic rat muscles, since the competitive potency of adrenergic drugs against [3H]prazosin binding was in the order prazosin much greater than phentolamine greater than (+/-) noradrenaline greater than yohimbine much greater than (+/-)-isoproterenol. The reduction of [3H]DHA binding sites was accompanied by an increase of an approximately equal amount in high-affinity [3H]prazosin binding sites. The Kd determined by kinetic analysis of [3H]prazosin binding was calculated from the ratio K-1/K1 that gave a value of 3.05 nM, which generally agreed with the 1.83 nM determined by saturation experiments (Scatchard plot). This phenomenon of a reduction in the beta-adrenoceptors and the occurrence of alpha 1-adrenoceptors in muscles during hypokalemia is discussed. alpha- and beta-adrenoceptors on soleus muscle membrane may play important but opposite roles in modulating potassium release from the muscle cells. PMID- 3026631 TI - [Detection and frequency of inapparent infections in epidemic foci of hepatitis A]. PMID- 3026630 TI - [Risk of teratogenic infections during pregnancy in mothers in Prague]. PMID- 3026632 TI - [Leukotrienes--mediators of inflammation, ischemia and shock. Their significance in the pathogenesis of central nervous system diseases]. PMID- 3026633 TI - Cell migration velocities in the crypts of the small intestine after cytotoxic insult are not dependent on mitotic activity. AB - The role of mitotic activity in the normal process of intestinal epithelial cell migration was investigated. The movement of [3H]TdR-labelled cells in the crypt villus column was used to study migration both in the crypts and on the villi. Radiation alone or in conjunction with other cytotoxic agents (hydroxyurea, cyclophosphamide and isopropyl-methane sulphonate) was used to eliminate cell division activity and to decrease crypt cellularity. This was done in order to determine the role of 'mitotic pressure' in driving cell migration. It has been clearly demonstrated in this study that cell migration, both within the crypts and on the villi, can take place in the complete absence of mitotic activity and after a drastic decrease in crypt cellularity. These results add to the continually mounting evidence against the idea that the 'pressure' generated by mitoses within the crypt or indeed in other epithelial regions is responsible for propelling epithelial cells. The data also demonstrate that the migration mechanisms are resistant to cytotoxic exposure. PMID- 3026634 TI - Fibronectin-degrading proteases from the membranes of transformed cells. AB - The local degradation of fibronectin substrata by Rous sarcoma virus-transformed chick embryonic fibroblasts requires cell-contact-related metalloendoprotease and serine-protease activities. Using fibronectin-containing SDS gels, two large proteases with apparent molecular weights of 120K and 150K were found only in the membrane fraction of transformed cells and were absent in normal cells. Both 120K and 150K proteases were active at neutral pH, but showed preferential inhibitor sensitivities of serine and metal proteases, respectively. The 150K protease appeared to account for most of the proteolytic activity since metalloendoprotease inhibitors completely blocked proteolytic activity of the 150K in fibronectin gels, more than 80% of the fibronectin-degrading activity of solubilized membranes, and largely suppressed the appearance of fibronectin degradation spots in cultures of transformed cells. PMID- 3026635 TI - Nuclear reconstitution in vitro: stages of assembly around protein-free DNA. AB - We have developed a cell-free system derived from Xenopus eggs that reconstitutes nuclear structure around an added protein-free substrate (bacteriophage lambda DNA). Assembled nuclei are morphologically indistinguishable from normal eukaryotic nuclei: they are surrounded by a double membrane containing nuclear pores and are lined with a peripheral nuclear lamina. Nuclear assembly involves discrete intermediate steps, including nucleosome assembly, scaffold assembly, and nuclear membrane and lamina assembly, indicating that during reconstitution nuclear organization is assembled one level at a time. Topoisomerase II inhibitors block nuclear assembly. Lamin proteins and membrane vesicles bind to chromatin late in assembly, suggesting that these components do not interact with chromatin that is formed early in assembly. Reconstituted nuclei replicate their DNA; replication begins only after envelope formation has initiated, indicating that envelope attachment may be important for regulating replication. PMID- 3026636 TI - Disassembly of the nucleus in mitotic extracts: membrane vesicularization, lamin disassembly, and chromosome condensation are independent processes. AB - We describe a stable cell-free mitotic extract derived from Xenopus eggs that contains activities necessary for nuclear envelope breakdown and chromosome condensation during mitosis. Using these cell-free extracts, we have demonstrated that nuclear envelope vesicularization, lamina solubilization, and chromosome condensation are independent and separable biochemical processes. We present evidence indicating that during mitosis nuclear membrane breakdown may involve the binding of a coating protein, lamin solubilization is enzymatically driven, and chromosome condensation involves both binding proteins and enzymatic activities including topoisomerase II. These results provide a coherent framework for investigating structural modification of the nucleus during mitosis at the biochemical level. PMID- 3026637 TI - Retrovirus antigens recognized by cytolytic T lymphocytes activate tumor rejection in vivo. AB - We have initiated the molecular definition of the antigens recognized by Gross MuLV-specific cytolytic T lymphocytes on the surface of Gross MuLV-induced tumor cells. A panel of target cells was obtained by the double transfection and expression of a retrovirus gene and a foreign H-2 gene in recipient mouse fibroblasts. Our results show that class I H-2 transplantation antigens have a directive influence in determining the antigenicity of proteins encoded by the gag and env MuLV genes. Genes not linked to H-2 influence the intensity and the specificity of the cytolytic T lymphocyte response to Gross MuLV-induced tumors. Finally, MuLV-induced antigens expressed by transfected fibroblasts induce tumor immunity and lead to accelerated tumor rejection in vivo. PMID- 3026639 TI - Steroid-dependent interaction of transcription factors with the inducible promoter of mouse mammary tumor virus in vivo. AB - We have used exonuclease protection in vivo as an assay to detect interaction of nuclear factors with the steroid-inducible promoter of mouse mammary tumor virus. Binding of two factors is detected uniquely at the steroid-activated promoter, and results in protection of sequences between -82 and approximately +12 One factor is identified as the murine homolog of nuclear factor 1. The second (designated factor i) binds downstream of nuclear factor 1 and protects sequences extending over the cap site. Binding activities associated with both factors can be detected in crude nuclear extracts; their apparent concentrations are unaffected by hormone treatment of the cells. These results demonstrate that glucocorticoid induction of transcription results from receptor-mediated establishment of a transcription factor complex at the promoter rather than activation of a preexisting complex. PMID- 3026638 TI - Involvement of common and cell type-specific pathways in c-fos gene control: stable induction of cAMP in macrophages. AB - The c-fos proto-oncogene is rapidly and transiently induced by PDGF in fibroblast and by CSF-1 in macrophages. In both cells, the breakdown of phospholipids with the ensuing activation of protein kinase C (PKC) and intracellular release of Ca2+ seems to play a role in the induction of c-fos. The transient induction of c fos mRNA and protein by PDGF is both increased and prolonged by inhibitors of calmodulin, apparently by inhibiting the degradation of c-fos mRNA. While no response to cyclic nucleotides is observed in fibroblasts, cAMP is a strong inducer of c-fos in macrophages. In contrast to the transient induction by PKC/Ca2+, cAMP induces stable transcription of the c-fos gene for many hours, suggesting the existence of different mechanisms regulating c-fos transcription in the same cell. PMID- 3026640 TI - Stimulation of a Chlamydomonas chloroplast promoter by novobiocin in situ and in E. coli implies regulation by torsional stress in the chloroplast DNA. AB - We have characterized regulation of a complex Chlamydomonas reinhardtii chloroplast (PA) whose activity is stimulated by the DNA gyrase inhibitor novobiocin, both in the alga itself and in a heterologous Escherichia coli plasmid system. Since novobiocin is known to reduce torsional stress in E. coli DNA, we interpret our results to mean that PA is regulated by torsional stress in the chloroplast DNA. In E. coli, where we could readily manipulate PA, we found that this regulation depends on sequences upstream of PA. These sequences contain at least two different kinds of silencing elements that inhibit PA in the absence of novobiocin. Novobiocin stimulates PA only when the promoter-distal silencing element is present. PMID- 3026641 TI - Topological characterization of the simian virus 40 transcription complex. AB - We have used sedimentation analysis as well as agarose gel electrophoresis to characterize the topological state of the DNA of the Simian Virus 40 (SV40) transcription complex. We found that the complex DNA contained constrained topological tension, presumably resulting from nucleosome-like structures, but no detectable unconstrained (i.e., relaxable) topological tension. These results contradict previous conclusions that the SV40 transcription complex contains only unconstrained topological tension. Our findings are also the opposite of what has been proposed to be the case for the 5S gene analyzed in Xenopus oocytes. Thus the proposal that expression from the 5S gene is associated with substantial topological tension is not valid for expression from the SV40 late gene. PMID- 3026642 TI - SV40 small t antigen enhances the transformation activity of limiting concentrations of SV40 large T antigen. AB - A murine recombinant Neo(r) retrovirus encoding the SV40 small t antigen was used to infect Balb/c 3T3 CIA31 cells. From analyses of G418-resistant clones containing at least as much intact t as Cos-1 cells, we found that t, alone, had no detectable A31 transforming activity. In contrast, we noted that SV40 large T promoted A31 agar colony formation when present over a 5- to 7.5-fold concentration range. However, at the low end of the spectrum, its transforming effect was manifest inefficiently except in the presence of t. Thus a major role for t in the SV40 transforming mechanism is to enhance directly or indirectly the transforming function of T. PMID- 3026644 TI - Molecular analysis of the Duchenne muscular dystrophy region using pulsed field gel electrophoresis. AB - Genetic and molecular studies show that the Duchenne muscular dystrophy (DMD) locus at Xp21 is large and complex. We have analyzed this region using pulsed field gel electrophoresis (PFGE) and have determined physical distances between Xp21 probes. The sum of the sizes of the Sfil restriction fragments detected by these probes is greater than 4000 kb. The deletion endpoints in two DMD patients were detected by observing changes in these restriction fragments. In addition, the Xp21 breakpoint for the X;1 translocation in an affected female was mapped. These results demonstrate the applicability of PFGE for analysis of Xp21, and should facilitate the mapping of other translocations and deletions in this region, some of which lead to glycerol kinase deficiency and adrenal hypoplasia as well as DMD. PMID- 3026643 TI - Evidence for aberrant activation of the interleukin-2 autocrine loop by HTLV-1 encoded p40x and T3/Ti complex triggering. AB - In this study we provide evidence that distinct DNA sequences within the 5' flanking regions of the genes for interleukin-2 (IL-2) and its receptor (IL-2R) are involved in human T-cell-specific activation of transcription by p40x, a product of human T cell leukemia virus type I (HTLV-1). The same DNA sequences appear to be responsible for induction of the genes in a T cell line, Jurkat, by mitogens. Although the IL-2 gene sequences are activated by p40x with much lower efficiency than the IL-2R gene sequences, they are synergistically activated by the p40x expression and subsequent extracellular stimulation by Concanavalin-A or anti-T3. We propose a model for two-step activation of the IL-2 autocrine loop in ATL development. PMID- 3026645 TI - Genetic analysis of the E. coli division clock. PMID- 3026646 TI - The transcription complex of vesicular stomatitis virus. PMID- 3026647 TI - Ability of the hydrophobic fusion-related external domain of a paramyxovirus F protein to act as a membrane anchor. AB - The hydrophobic NH2 terminus of F1 (FRED) of the simian virus 5 fusion (F) protein is implicated in mediating cell fusion, but in the inactive F0 precursor the FRED is translocated across membranes. Hybrid proteins containing the FRED as a potential membrane anchorage domain and a mutant of F0 lacking the preceding five-arginine cleavage/activation site were used to study the effect of position on the FRED. The experiments indicate that the SV5 F protein has evolved an exquisite control system for biological activity: the FRED is close to the threshold of hydrophobicity required to function as a membrane anchor. The FRED is not sufficiently hydrophobic to halt translocation when in an internal position, but on cleavage/activation the threshold of hydrophobicity is effectively lowered, and the FRED, now the NH2 terminus of F1, is able to interact stably with membranes. PMID- 3026648 TI - The c-Ha-ras oncogene and a tumor promoter activate the polyoma virus enhancer. AB - A c-Ha-ras oncogene, to a lesser extent the c-Ha-ras proto-oncogene, and the tumor promoter 12-O-tetradecanoylphorbol-13-acetate activate the inactive polyoma virus (Py) enhancer in a myeloma cell line and the partially active Py enhancer in NIH 3T3 fibroblasts, but have no effect on the active Py enhancer in LMTK- fibroblasts. In addition, c-Ha-ras can stimulate the inactive Py enhancer in embryonal carcinoma F9 cells. c-Ha-ras activation in embryonal carcinoma cells does not appear to involve reversal of "E1A-like" inhibition of the enhancer. We suggest that modulation of cellular enhancer activity could play a key role in tumorigenesis by oncogenes. PMID- 3026649 TI - In vivo effects of catecholamines and glucocorticoids on mouse thymic cAMP content and thymolysis. AB - In vivo administration of the beta-adrenergic agonist, isoproterenol, induced a rapid, dose-dependent increase in the mouse thymic cyclic AMP (cAMP) content. Hydrocortisone (1 mg/animal), at a concentration which by itself did not alter the cAMP content of the thymus, markedly potentiated the effect of isoproterenol (5 micrograms/animal). Isoproterenol or hydrocortisone treatment led to a significant decrease in thymic weight and an even greater decrease in thymocyte number. In addition, the simultaneous administration of both agents produced additive effects on thymic atrophy. It appears from these results that glucocorticoids and catecholamines exert a negative control on the thymic size by increasing the programmed cell death of some cell subpopulations. Thus, glucocorticoids and catecholamines, either alone or in association, may influence the immune system under physiological or pathophysiological conditions. PMID- 3026650 TI - Radiation leukemia virus-transformed immunocompetent T cells. II. Antigen-induced macrophage migration inhibition factor and leukocyte migration inhibition factor production. AB - OVA-specific T cells were immortalized by infection with radiation leukemia virus (RadLV). Some clones derived from such population were shown to exhibit helper activity. We then tested clones without such function and found among them some that secreted macrophage migration inhibition factor (MIF) and leukocyte migration inhibition factor (LIF) upon exposure to the antigen in vitro. The lymphokine-producing clones, which were Thy-1+, Ly-1+ and Ly-2-, did not secrete MIF and LIF constitutively. Like other antigen-specific T cells, the immortalized clones could not be stimulated by free soluble antigen but required macrophages for presentation and for triggering the lymphokine production. The antigen activated clones exclusively produced MIF and LIF, but not interleukin 2 or colony-stimulating factor. They neither provided helper activity nor induced delayed-type hypersensitivity. The data suggest that the T-cell clones carry the antigen receptors and that their antigen-inducible biological function is restricted to the migration inhibitory factor production. PMID- 3026652 TI - Allogeneic responses of human fetal (22- to 25-week) peripheral blood lymphocytes: preferential recruitment of cytotoxic effector cells with both CTL activity and NK-like function. AB - In the present study, we have characterized human cytotoxic effector lymphocytes generated following in vitro immunization of normal fetal (22- to 25-week) peripheral blood mononuclear cells (FPBMC) by an allogeneic Epstein-Barr virus transformed B-cell line termed LAZ388. Primary stimulations led to strong FPBMC proliferation. However, subsequent addition of LAZ388 cells to the cultures on Day 8 did not trigger conventional secondary responses. In fact, further proliferation of activated FPBMC required the addition of exogeneous interleukin 2. Cytotoxic activity generated in the mixed-lymphocyte reactions was assayed against LAZ388 immunizing cells as well as against the highly susceptible natural killer (NK) target cell line K562. Eight days after stimulation by LAZ388, there was no specific lysis and a moderate NK-like activity. However, following second and subsequent stimulations a strong killing was measured against both LAZ388 and K562 cells. Blocking experiments performed with relevant monoclonal antibodies suggested that cytotoxicity against immunizing cells was conventionally directed at MHC gene products. Effector cells were further studied using cloning procedures; it was found that all cloned cell lines able to kill LAZ388 cells were also strongly active against K562. Both types of cytotoxic function appeared to be mediated via surface receptors physically or at least functionally associated with T3 proteins. PMID- 3026651 TI - Differential activation of cytotoxic responses by Burkitt's lymphoma (BL)-cell lines: relationship to the BL-cell surface phenotype. AB - Epstein-Barr (EB) virus-positive Burkitt's lymphoma (BL) cell lines, recently established from tumour biopsies and displaying chromosomal translocations indicative of their malignant origin, can be classified into two broad sets: (i) lines growing predominantly as single cells/small clumps whose cell surface markers remain close to those of the original tumour cells, and (ii) lines whose growth pattern and cell surface markers have progressed closer to the more "lymphoblastoid" phenotype displayed by all in vitro transformed B-cell lines (LCLs) of normal origin. When compared to LCLs derived from normal B cells of the same patient, BL-cell lines in set (i) generally showed a lower expression of HLA class I and class II antigens and a reduced ability to activate both allospecific and nonspecific (natural killer-like) cytotoxic responses when cocultured with peripheral blood lymphocytes. By contrast, the HLA antigen expression and in vitro stimulatory capacity of most BL-cell lines in set (ii) were much closer to the values displayed by their corresponding LCLs. Since set (i) rather than set (ii) BL cell lines are phenotypically representative of the malignant cells as they exist in vivo, this work suggests that successful outgrowth of the virus carrying tumour cells in the affected host may be facilitated by the inability of these cells to stimulate strong cytotoxic responses. PMID- 3026653 TI - 5'-nucleotidase activity of murine T-cell subpopulations separated according to their Lyt2 and L3T4 phenotypes. AB - Murine T lymphocytes were separated by "panning" into four subpopulations, according to their Lyt2 and L3T4 phenotypes: Lyt2+L3T4+, Lyt2-L3T4-, Lyt2+L3T4-, and Lyt2-L3T4+. The activity of ecto-5'-nucleotidase in each subpopulation was measured. 5'-Nucleotidase activity was undetectable in Lyt2+L3T4+ cortical cells but was expressed in medullary Lyt2-L3T4+ and Lyt2+L3T4- T lymphocytes. The small cortical subpopulation of thymocyte precursors with the Lyt2-L3T4- phenotype expressed levels of 5'-nucleotidase comparable to the levels of medullary, mature lymphocytes. These results suggest that the use of ecto-5'-nucleotidase as a marker of the degree of T-cell maturation is questionable. PMID- 3026654 TI - Opioid peptides modulate production of interferon gamma by human mononuclear cells. AB - The opioid neuropeptides have previously been shown to bind to and affect leukocyte function including lymphocyte proliferation, NK-cell activity, mononuclear cell chemotaxis, immunoglobulin synthesis, and lymphokine production. The effect of the opioid peptides beta-endorphin and Met-enkephalin on interferon gamma (IFN) production by concanavalin A-stimulated human mononuclear cells was examined. Both beta-endorphin and Met-enkephalin enhanced IFN production by the majority of donor mononuclear cells tested and did so at concentrations between 10(-14) and 10(-10) M. When 10(-12) M beta-endorphin or Met-enkephalin were included in concanavalin A-stimulated mononuclear cell cultures, IFN concentrations were significantly enhanced to 205 +/- 45 and 252 +/- 67% of control, respectively. Although the majority of cell preparations tested exhibited an enhanced production of IFN in response to these opioid peptides, some did not. When beta-endorphin or Met-enkephalin were utilized at 10(-11) M, 10 of 15 and 7 of 11 responded with IFN production greater than 20% above the control (untreated) level. There was not an absolute correlation between an enhanced response to beta-endorphin and Met-enkephalin, suggesting the presence of multiple receptor types on these cells for opioids. The opioid receptor antagonist, naloxone, did not significantly prevent the opiate effect. When 10( 8) M naloxone was included in cultures containing 10(-12) M beta-endorphin or Met enkephalin no significant inhibition of the effect of either opioid on IFN production was observed. PMID- 3026655 TI - Macrophage membrane proteins: possible role in the regulation of priming for enhanced respiratory burst activity. AB - Brief exposure of macrophages to the proteolytic enzymes papain, elastase, or trypsin primed them for enhanced production of superoxide anion (O2-) in response to stimulation by phorbol myristate acetate (PMA). Priming by trypsin was achieved at 0 degree C, at which temperature trypsin functions as a protease but is not internalized, supporting the concept that protease priming depends on modification of the plasma membrane. Analysis of external membrane proteins after radioiodination of intact cells and separation by gel electrophoresis indicated that papain treatment of macrophages resulted in the cleavage of a membrane protein with a molecular weight of approximately 305K. Membranes from macrophages primed by elicitation with Corynebacterium parvum also demonstrated a reduced amount of the membrane protein at approximately 305 kDa, as well as a reduction of a protein at about 270 kDa. Lipopolysaccharide-elicited macrophages showed a reduced amount of a protein at about 175 kDa. Continuous spectrophotometric assays of O2- release from adherent macrophages indicated that after exposure to a stimulus, protease-treated cells produced O2- more quickly than did control cells (reduced lag time). Inhibitors of protein synthesis augmented the priming effect of papain when added with the protease. These results suggest that protease-induced priming results from inactivation of a membrane protein (or proteins) that exerts a down-regulating effect on the respiratory burst. PMID- 3026656 TI - Macrophage activation in rat models of inflammation and arthritis: determination of markers of stages of activation. AB - Disease-associated alterations in macrophage functions were assessed by investigating the stages of activation of peritoneal macrophages obtained from adjuvant-induced arthritic rats. The stages of activation were established by defining several functional parameters in macrophages obtained from normal, sterile-irritant injected and Propionibacterium acnes injected animals. Peritoneal macrophages taken from arthritic rats 17 days post adjuvant injection displayed parameters characteristic of activated, but not elicited or resident macrophages. Specifically, an increased number of macrophages was recovered from arthritic rats which spread readily in culture, exhibited enhanced Fc receptor mediated phagocytosis, increased leucine aminopeptidase ectoenzyme activity, enhanced secretion of prostaglandin E2 and interleukin 1, and ability to lyse tumor cells spontaneously. In addition, these macrophages were impaired in their ability to secrete superoxide anion. These data demonstrate distinct differences in parameters of peritoneal macrophage activation in rats compared to mice and that macrophage activation is associated with disease progression in adjuvant induced arthritic rats. PMID- 3026657 TI - Regulation of the growth of Epstein-Barr virus-infected B cells: temporal profile of the in vitro development of three distinct cytotoxic cells. AB - The time course of the appearance of cytotoxic cells was examined in cocultures of E-rosetting (E+) cells and EBV-infected non-T cells (4:1 ratio) from the blood of VCA-positive healthy adults. Classical HNK-1+ NK cells were present at the initiation of the cultures and they produced 76 +/- 2% specific 51Cr-release from K-562 cells, but they did not effectively lyse the NK-resistant Daudi cells, nor did they kill autologous EBV-induced lymphoblasts (LCLEBV). The NK activity decreased during the first week in culture to 40 +/- 7% cytotoxicity. At the same time, nonspecific cytotoxic cells capable of killing Daudi as well as K562 developed and persisted into the third week in culture when it declined. This later nonspecific cytotoxicity was mediated by 4F2+, T8-, HNK-1- activated E+ cells. After 10 days in culture, killing of autologous LCLEBV increased continuously, from 4 +/- 3% at Day 10 to 38 +/- 4% by Day 22. The cytotoxicity to LCLEBV was mediated by classical T8+ CTL, and it was antigen specific and at least partially HLA Class I restricted. The regression of BEBV growth that occurs in E+/BEBV cocultures coincides with the development of this CTL-mediated cytotoxicity. PMID- 3026658 TI - Elevation of cyclic 3'5' adenosine monophosphate levels by cholera toxin inhibits the generation of interleukin 2 activity. AB - Several molecules can interact with membrane receptors on mononuclear cells to increase intracellular levels of cyclic adenosine monophosphate (cAMP). We used the cholera toxin (CT), a cAMP elevating agent, to study the influence of this nucleotide on the production of interleukin 2 (IL-2) by human peripheral blood mononuclear cells stimulated by phytohemagglutinin and phorbol myristate acetate. Stimulated generation of IL-2 activity was inhibited by CT but not by its B subunit. The inhibition was potentiated by addition of theophylline. Therefore the synthesis and/or release of IL-2 is controlled by intracellular cAMP levels and may be modulated by agents active on this nucleotide system, such as bacterial toxins, glycoprotein hormones, or neurotransmitters. PMID- 3026659 TI - VSV replication in normal and transformed T cells, an assay for T suppressor cell activation. AB - An infectious center viral plaque assay has been utilized to quantitate activated T suppressor (Ts) cells. This assay is based on two observations. Namely, resting T cells do not serve as good replicative hosts for many viruses, including vesicular stomatitis virus (VSV), and that Ts cells can be enriched by their ability to bind to antigen-coated dishes. Our data show that Ts cells specific for either the TNP hapten or for dextran will replicate VSV upon antigenic and/or mitogenic activation, whereas resting Ts and hapten-specific B cells are less efficient in this process. This system will now allow the direct quantitation of Ts cells and their activation properties. PMID- 3026660 TI - 1 alpha, 25-dihydroxyvitamin D3 modulates the growth of 3T3 cells and human skin fibroblasts stimulated by platelet-derived growth factor. AB - We investigated the effect of 1 alpha,25-dihydroxyvitamin D3 (1,25 (OH)2 vit D3) on the 3H-thymidine uptake by Balb/c 3T3 cells and by human skin fibroblasts stimulated by normal human serum or by purified PDGF. We found an inhibitory effect of 1,25 (OH)2 vit D3 on the DNA synthesis of Balb/c 3T3 cells grown in the presence of human serum as well as in the presence of PDGF. At 5% human serum this effect is minimal at 10(-12) M 1,25 (OH)2 vit D3 and is maximal at 10(-9) M. On the DNA synthesis of human fibroblasts stimulated by human serum or by PDGF a modulatory effect of 1,25 (OH)2 vit D3 was shown. On these cells the vitamin had a stimulatory effect between 10(-11) and 10(-9) M and an inhibitory effect at very high concentrations (10(-7) M). Our results suggested that the effect of 1,25 (OH)2 vit D3 on fibroblast DNA synthesis could be mediated by interactions with its specific intracellular receptor. 1,25 (OH)2 vit D3 had no any action on the growth of human fibroblasts stimulated by fibroblast growth factor. PMID- 3026661 TI - Growth inhibitory factor diffusing from chick embryo fibroblasts. AB - Density-dependent inhibition (DDI) of growth is assumed to be the result of diffusion in the medium of growth inhibitory molecules. In this work, we demonstrate the presence of inhibitory molecules (IDFc: chicken inhibitory diffusible factor) in the medium of chick embryo fibroblasts (CEF) cultures. IDFc partially purified by Bio-Gel P150 chromatography followed by reverse phase FPLC. The dose-response curve showed that 250 ng/ml IDFc inhibited 50% DNA synthesis. IDFc was also able to inhibit the growth of sparse cultures of CEF; this inhibition was reversible. IDFc was unable to prevent the DNA synthesis in cells transformed by v-src gene expression. These results suggest that IDFc is involved in the DDI of CEF growth. PMID- 3026662 TI - Adenosine potentiates the delaying effect of dbcAMP on meiosis resumption in denuded mouse oocytes. PMID- 3026663 TI - [Incidence of rotaviruses in the feces of neonates determined by the ROTA-ELISA Bioveta immunoenzyme test kit]. PMID- 3026665 TI - [Levels of adenosine cyclic 3',5'-monophosphate in umbilical cord blood and maternal blood]. PMID- 3026664 TI - [Laboratory diagnosis of infectious mononucleosis]. PMID- 3026666 TI - [Protection of the lens against ionizing radiation injury using gammaphos]. PMID- 3026667 TI - [Neuropathy caused by vincristine]. PMID- 3026669 TI - [Haemostatic effect of crude crystals of processed human hair]. PMID- 3026668 TI - Adriamycin, cyclophosphamide and etoposide (ACE) in the treatment of small cell lung cancer. AB - Thirty-five small cell lung cancer (SCLC) patients were treated with combination chemotherapy including adriamycin, cyclophosphamide, etoposide (ACE). Out of 32 evaluable patients there were 21.9% complete responses and 53.1% partial responses with an overall median survival of 37 weeks (50 weeks for patients with limited disease and 34 weeks for patients with extended disease). Toxicity was generally well tolerated. In conclusion the ACE regimen results in being active and safe in the treatment of SCLC. PMID- 3026670 TI - Diagnostic histopathologic criteria in pediatric tumors: an eclectic review. AB - Malignant tumors of infants and children, unlike their adult counterparts, are commonly undifferentiated or show minimal evidence of differentiation. Because of this, many of these tumors pose a formidable dilemma to the general pathologist in distinguishing one from another. A broad sampling of the more common malignant neoplasms and of newly recognized histologic subtypes which are restricted more or less to the pediatric age group is presented. The minimal microscopic criteria required to establish a diagnosis are given using widely accepted pathologic classifications and seminal journal references as a guide. Ancillary microscopic features, helpful histochemical stains, immunohistochemistry, and ultrastructural morphology are described for most tumors. Histologic distinction from diagnostic mimickers is discussed. Where appropriate, histologic grading and tumor subclassification are presented and their prognostic relevance is reviewed. PMID- 3026672 TI - [Retroviruses]. PMID- 3026671 TI - [Diarrhea caused by Aeromonas in Ivory Coast]. AB - The authors drawn epidemiological, clinical conclusions, and the geographical data concerning the diarrheas with Aeromonas in Ivory-Coast. The results relate to a study realized from January 1982 to July 1985. Out of the 1,594 excrements 627 germes have been isolated and among which 153 strains of aeromonas if for instance 24.4% of the whole germs. Aeromonas caviae is the most frequent species (50%) compared to 27.3% for Aeromonas sobria, and 22.7% for Aeromonas hydrophila. PMID- 3026673 TI - Single-radial-haemolysis test for diagnosing flavivirus infections, particularly Japanese encephalitis. AB - Use of the single-radial-haemolysis (SRH) technique for the diagnosis of flavivirus infections is described. A large number of paired and single convalescent serum samples collected from cases of encephalitis during two major outbreaks in Kolar district of Karnataka State in India during 1977 and 1979 were tested by this technique. The results were compared with those obtained in the haemagglutination inhibition (HI) test in all cases, and the complement fixation (CF) and neutralization tests in some cases. Japanese encephalitis virus was shown by the SRH test to be the major etiologic agent responsible for both epidemics. This was corroborated by the HI, CF and neutralization test results. The single-radial-haemolysis test was found to be simpler and more specific and sensitive than the haemagglutination inhibition test. PMID- 3026674 TI - Enhancement of etoposide-induced cytotoxicity by cyclosporin A. AB - Following the clinical observation of enhanced antineoplastic action of etoposide in the presence of cyclosporin A (CyA), we investigated this drug interaction in several in vitro and in vivo tumor systems. Macromolecular DNA damage induced by etoposide at drug levels comparable to plasma AUC values achieved in patients was increased not only in leukemic peripheral blood cells from patients but also in mononuclear peripheral blood cells from a healthy donor. Intracellular retention of radioactivity from 3H-etoposide was increased by a factor of 1.5 at the most in the presence of CyA. The cytotoxicity of etoposide and adriamycin to L 1210 leukemic cells was clearly enhanced, whereas CyA had no effect on the action of cisplatin or ionizing irradiation. At CyA blood levels not exceeding 1.44 microgram/ml, increased tumor inhibition of etoposide was observed in a human embryonal cancer xenograft, but there was also higher lethality in normal mice. We conclude from our own data and from other recent findings that with respect to chemosensitization the effects of CyA resemble those of calcium channel blockers or anticalmodulin agents. In contrast to calcium channel blockers, however, adequate plasma levels of CyA can well be achieved in patients. PMID- 3026675 TI - The effect of verapamil on the pharmacokinetics of adriamycin. AB - The concurrent administration of adriamycin (intravenous) and verapamil (oral) is of considerable interest because of experimental data suggesting that resistance to adriamycin may be overcome by this means. The potential for a pharmacokinetic interaction between the two drugs has therefore been investigated in five patients with small cell lung cancer treated with combination chemotherapy comprising adriamycin, VP16, vincristine and cyclophosphamide. The data indicate that a significant interaction takes place. Adriamycin peak levels, terminal half life and the volume of distribution at steady state are higher, whereas plasma drug clearance and the volume of the central compartment are lower with co administration of verapamil. There was no evidence of enhanced drug toxicity in this study; however, the data should be considered in the interpretation of clinical trials in which adriamycin and verapamil are used together, both in terms of toxicity and tumour response. PMID- 3026677 TI - Incidence of cardiac septal defects in children with Wilms' tumour and other malignant diseases. AB - In a population-based series of 8045 children with malignant neoplasms in Great Britain, the incidence of septal defects was 0.40%. The high rate of 19% in Down's syndrome children and the overall rate of 0.28% in children without Down's syndrome were both comparable with rates found in previous large prospective studies. The incidence of septal defects in children with Wilms' tumour was 1.82%, greater than 10 times that for non-Down's children with other neoplasms. The presence of this association in Wilms' tumour patients with and without aniridia and the recent finding of a loss of heterozygosity in chromosome 11 in many Wilms' tumours taken from patients without aniridia suggest the possibility of a link between Wilms' tumour, some septal defects and chromosomal abnormalities. PMID- 3026676 TI - DNA sequences in human nasopharyngeal carcinoma cells that specify susceptibility to tumor promoter-induced neoplastic transformation. AB - Homologs of the recently described mouse pro genes, that transfer sensitivity to promotion of neoplastic transformation by tumor promoters, have been cloned from a genomic library of the human nasopharyngeal carcinoma (NPC) cell line CNE2. Both pro-1 and pro-2 homologs were identified by screening this library with mouse probes, but only the pro-1 homologs were able to confer sensitivity to TPA induced transformation when transferred to promotion-insensitive mouse JB6 cells. This suggests a possible role for the putative pro-1 gene in the etiology of human NPC. PMID- 3026678 TI - Induction of thymomas by N-methyl-N-nitrosourea in AKR mice: interaction between the chemical carcinogen and endogenous murine leukaemia viruses. AB - AKR mice develop thymomas spontaneously when greater than 6 months old but when young AKR mice are treated with N-methyl-N-nitrosourea (MNU) they develop thymomas at 3-6 months of age. In this study the potential role of oncogene activation in the development of both the spontaneous and MNU-induced thymomas in AKR mice has been examined by DNA transfection into NIH3T3 mouse fibroblasts and by Southern analysis of tumour DNA. The results show that a high proportion of MNU-induced thymomas contain activated cellular rasK while no activated cellular ras genes were detected in spontaneous thymomas. Southern analysis of tumour DNA revealed that 2/30 spontaneous tumours and 2/52 MNU-induced tumours contained alterations in the c-myc gene while 5/29 spontaneous tumours and 6/56 MNU-induced tumours contained alterations in the Pim-1 gene. A more detailed analysis of the Pim-1 gene demonstrated that the alterations observed in most MNU-induced and spontaneous tumours resulted from proviral integration at the 3' end of this gene. Our analyses also demonstrated that the majority of MNU-induced tumours, including those containing rearrangements in the Pim-1 gene, lacked the somatically acquired recombinant MCF proviruses that are present in most spontaneous AKR lymphomas. These results provide evidence that the mechanisms of development of MNU-induced and spontaneous tumours in AKR mice are distinct and the development of thymomas that contain proviral integrations at the Pim-1 locus in the MNU-treated AKR mice involve cooperation between the chemical carcinogen and endogenous murine leukaemia viruses. PMID- 3026679 TI - Induction of cellular functions in spontaneously immortalized rat-2 cells transfected with cloned herpes simplex virus type 2 (HSV-2) DNA. AB - Experiments were done to determine if cloned transforming sequences from herpes simplex virus type 2 (HSV-2) DNA confer upon transfected Rat-2 cells the capacity to be stimulated in phospholipase and cyclooxygenase activities following 12-O tetradecanoylphorbol-13-acetate (TPA) treatment. Tumor-derived Rat-2 cells transformed with sub-fragments (BamHI-E, HindIII/HpaI-ED, or PstI-C) overlapping the right-hand end of the BglII-C transforming region of HSV-2 DNA were stimulated by TPA in both phospholipase activity, measured by deacylation of arachidonic acid, and cyclooxygenase activity, measured by prostaglandin synthesis. Non-transfected Rat-2 control cells showed no increase in these enzyme activities following TPA treatment. To our knowledge, this is the first evidence that cloned HSV-2 DNA has the capacity to induce cellular functions (phospholipase and cyclooxygenase) in transformed mammalian cells. PMID- 3026680 TI - Induction kinetics of mutations at two genetic loci, DNA damage and repair in CHO cells after different exposure times to N-ethyl-N-nitrosourea. AB - Ouabain resistance (ouar) and 6-thioguanine resistance (6-TGr) mutation frequencies were measured in Chinese hamster ovary cells after treatment with N ethyl-N-nitrosourea (ENU) for varying periods of time. Maximal mutation frequency at the Na+/K+ ATPase gene locus (ouar mutations) was attained within 5 min of exposure, whereas the mutation frequency at the hypoxanthine guanine phosphoribosyltransferase locus (6-TGr mutations) continued to increase up to 60 min, following the theoretical curve for exponential decay of ENU with time. Detection of DNA single strand breaks (ssb) by alkaline elution showed that maximal levels were attained within 5 min of treatment with ENU. Fast repair of DNA ssb occurred early after exposure (greater than 50% repair within 10 min). Analysis of DNA ethylation products by h.p.l.c. showed initially rapid removal of O2-ethylcytosine (25% in the first hour), slow removal of 7-ethylguanine, 3 ethyladenine and 3-ethylguanine and no removal at all of O6-ethylguanine, O4 ethylthymine and ethylphosphotriesters. These time-course studies reveal different target gene responses in the fixation of DNA damage into mutations. PMID- 3026681 TI - Inhibition by naloxone of neutrophil superoxide release: a potentially useful antiinflammatory effect. AB - The working hypothesis of many studies of shock has been that naloxone acts by blocking centrally and/or peripherally located opioid receptors. At plasma concentrations used to treat experimental shock (10(-6) M and above), naloxone inhibited the in vitro release of superoxide (O2-) by human neutrophils that were stimulated by the E. coli peptide N-formyl methionyl leucyl phenylalanine (FMLP). Superoxide release stimulated by phorbol 12,13-dibutyrate (PDB) was also inhibited by naloxone. Naloxone had no effect on the FMLP-stimulated release of beta-glucuronidase or lysozyme. Naloxone had no effect on 3H FMLP receptor binding. Studies utilizing 3H naloxone revealed the presence of a ligand-specific naloxone binding site on human neutrophils with a Kd of 1.2 X 10(-5) M, which is close to the ID50 of the inhibitory effect upon O2- release (1.8 X 10(-5). Thyrotropin releasing factor (TRF) had no effect upon 3H naloxone binding or on O2- release. Verapamil, a calcium channel blocker, inhibited 3H naloxone binding, and O2- release while nifedipine, another calcium channel blocker had no effect on either assay except at 10(-4) M, at which concentration 3H naloxone binding as well as the release of O2- were increased. These experiments suggest that the inhibitory effect of naloxone upon O2- release is mediated via a specific binding site. PMID- 3026682 TI - Sympathetic innervation alters growth and intrinsic heart rate of fetal rat atria maturing in oculo. AB - The influence of sympathetic innervation on the growth and intrinsic rate of beating established by fetal rat heart was studied by culturing fetal atrial tissue in sympathetically innervated and denervated anterior eye chambers of adult Sprague-Dawley rats. One anterior eye chamber in each host rat was sympathetically denervated by removing the ipsilateral superior cervical ganglion. In oculo, atrial grafts were vascularized by blood vessels sprouting from the iris and innervated by sympathetic and parasympathetic fibers from the ground plexus of the iris. Innervation was assessed by light-activated efferent nerve stimulation to the grafts that changed their rates of beating. The norepinephrine contents of 16 atria cultured for 2.5 months in sympathetically innervated and denervated eye chambers were 5.7 +/- 1.1 ng/implant vs. 0.2 +/- 0.07 ng/implant (mean +/- SEM), indicating permanent sympathetic denervation of the anterior eye chamber and the implanted atria. By 8 weeks in oculo, atria maturing in sympathetically innervated anterior eye chambers were 86% larger than those in denervated eye chambers (2.22 +/- 0.29 vs. 1.19 +/- 0.13 mm2); the weight of innervated transplants was over 3 times that of noninnervated grafts (2.35 +/- 0.75 vs. 0.76 +/- 0.21 mg). After implanted atria had ceased growing rapidly (2.5 months in oculo), bipolar electrodes were implanted adjacent to the cornea to record impulses from atrial grafts while host rats were unanesthetized. The dark-adapted baseline heart rates of sympathetically innervated and noninnervated atria were virtually identical (289 vs. 290 bpm). Graft intrinsic heart rate was estimated by combined beta-adrenergic and muscarinic receptor blockade with atenolol (1.0 mg/kg) and methylatropine (10 micrograms/kg). Sympathetically innervated transplants had lower intrinsic heart rates than noninnervated atria (134 +/- 25 vs. 213 +/- 12 bpm). These data suggest that sympathetic innervation of the developing heart influences both growth and intrinsic rate of beating. PMID- 3026684 TI - Variables affecting resolution of lung phospholipids in one-dimensional thin layer chromatography. AB - Resolution of the confusion in the literature about the separation of lung phospholipids in thin-layer chromatographic systems has awaited a systematic study of the variables that potentially affect this separation. In this study I show that: incorporation of ammonium sulfate into silica gel "GHL" has a dramatic effect on separation of lung phospholipids; this effect is equally dramatic but different in activated and nonactivated gels; when it picks up moisture, ammonium sulfate-activated gel very rapidly loses its ability to resolve lecithin from phosphatidylinositol; in gel containing ammonium sulfate, small amounts of phosphatidylethanolamine are hydrolyzed to lyso-phosphatidylethanolamine. PMID- 3026683 TI - The role of endogenous opioids in congestive heart failure: effects of nalmefene on systemic and regional hemodynamics in dogs. AB - We studied the role of endogenous opiates and their interrelationships with the sympathetic nervous system in an experimental preparation of right-sided congestive heart failure (CHF) produced by surgical tricuspid avulsion and progressive pulmonary arterial constriction. Three groups of dogs with CHF and one group of sham-operated dogs were studied. One group of dogs with CHF was given normal saline as pretreatment, while the other two groups were pretreated with either propranolol alone (beta-blockade) or propranolol plus prazosin (alpha plus beta-blockade). CHF was characterized by weight gain, ascites, elevated right atrial pressure, tachycardia, and reduced cardiac output. Compared with sham-operated animals, animals with CHF exhibited significantly higher baseline levels of plasma beta-endorphin and cortisol. Furthermore, only the animals with CHF responded to the opiate receptor-antagonist nalmefene with significant increases in plasma beta-endorphin, cortisol, and adrenocorticotropic hormone. Administration of nalmefene increased aortic blood pressure, cardiac output, left ventricular dP/dt and dP/dt/P, and blood flow to the myocardium, skeletal muscle, and kidneys in dogs with CHF, but had no appreciable effects in sham-operated dogs. beta-Receptor blockade abolished the increase in cardiac output, left ventricular performance, and blood flow produced by nalmefene, but had no effect on the pressor response to nalmefene. The increase in mean aortic pressure in the beta-blockade group was accompanied by an increase in skeletal muscle vascular resistance. Addition of prazosin in the alpha- plus beta-blockade group abolished the increases in mean aortic pressure and skeletal muscle vascular resistance, suggesting that the changes after propranolol probably resulted from unmasking of alpha-receptor-mediated vasoconstriction.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3026685 TI - Noninvasive evaluation of mineral content of bone without use of ionizing radiation. AB - The mineral content of bone can be quantified by recording a 31P NMR spectrum while the bone is stationary. The quantity of mineral in the bone is determined from the spectrum with a reference standard by comparison of relative peak areas. The phosphate of bone mineral is readily distinguished from inorganic phosphate and phosphorylated metabolites dissolved in cytosol and from the head groups of phospholipids in membranes. The technical feasibility of constructing a cost effective instrument for analysis of bone mineral content in the extremities is demonstrated. The possible utility of such a noninvasive assay for detecting osteoporosis and for monitoring the progress of treatment is discussed. PMID- 3026686 TI - Effect of gossypol on the activity of erythrocyte Na+-K+ ATPase activity in vitro. PMID- 3026687 TI - Adaptation of 5'-nucleotidase assay for use with the Olympus Demand. PMID- 3026688 TI - Phospholipase C and phosphatidylinositol phospholipase C. PMID- 3026689 TI - Competitive inhibitor binding assay (CIBA) of captopril and other ACE inhibitors. AB - We describe a new principle for measuring concentrations of pharmacologically active captopril or other angiotensin-converting enzyme (ACE) inhibitors in blood. Serum is incubated with 125I-labelled ACE inhibitor (351A, a p-hydroxy benzamidine derivative of N-(1-carboxy-3-phenylpropyl)-L-lysyl-L-proline) in a nonequilibrated system, in which label and ACE inhibitor compete for binding to added serum ACE. Free label is separated by adsorption to coated charcoal. Sensitivity for captopril is 2 ng/ml, and for other tested ACE inhibitors 0.25-5 ng/ml. Results in healthy volunteers showed rapid absorption of captopril with maximal concentration of active drug within 1 h, and fast disappearance within 2.5 h. Stability of captopril was improved by immediate 1:100 dilution of blood samples with assay buffer. In spite of this precaution, analysis should be performed within two days to avoid loss of active drug due to polymerization and protein binding. Samples of other tested ACE inhibitors can be frozen and later analyzed at convenience. The new principle is simple, sensitive, and specific. PMID- 3026690 TI - Effects of alpha-1 adrenergic blockade on the hormonal response to hypoglycaemic stress in normal man. AB - In order to evaluate the effect of alpha-1 adrenoreceptor regulation of ACTH release during insulin-induced hypoglycaemia, we studied the response to hypoglycaemia with and without prazosin premedication in eight normal men. Prazosin pretreatment did not affect basal or peak plasma ACTH, cortisol or GH during hypoglycaemic stress. However basal plasma levels of noradrenaline were increased (P less than 0.02) as were responses of AVP, angiotensin II (P less than 0.05) and noradrenaline (P less than 0.05) to hypoglycaemia after prazosin. Though it is possible that these augmented responses masked an inhibitory effect of prazosin, we were unable to demonstrate a major role for alpha-1 adrenergic receptors in mediating the ACTH response to hypoglycaemic stress in normal man. PMID- 3026691 TI - Clinical evaluation of a two-site immunoradiometric assay for adrenocorticotrophin in unextracted human plasma using monoclonal antibodies. AB - We have developed a sensitive two-site immunoradiometric assay (IRMA) for intact ACTH and its precursors, pro-opiomelanocortin and 22 kDa peptide in unextracted human plasma. The assay uses two monoclonal antibodies. Antibody 1A12, specific for ACTH 10-18, is radiolabelled and antibody 2A3 specific for the C-terminal region (ACTH 24-39), is coupled to Sephacryl S300 for the solid-phase. Samples are incubated for 18 h with labelled antibody followed by 2 h with solid-phase antibody. Separation employs the sucrose layering technique. Using human pituitary ACTH 1-39 (code 74/555) in diluent containing 10% horse serum to standardize the assay, the sensitivity (upper 99% confidence limit of zero standard) is 3.5 +/- 0.8 ng/l (n - 7). The mean coefficient of variation is 5.9% within-assay and 6.7% between-assay and is less than 10% between 22 and greater than 5000 ng/l. Mean recovery of ACTH 1-39 added to dexamethasone-suppressed human plasma is 109% and endogenous ACTH behaves indistinguishably from standard ACTH on dilution. In normal subjects, mean plasma ACTH levels are 30 ng/l at 0730 h, and 15 ng/l at 1630 h at rest. ACTH concentrations are between 60 and 330 ng/l, 8-10.5 h after metyrapone (2 g orally at 2300 h), between 140 and 320 ng/l, 30-60 min after insulin-induced hypoglycaemia, and less than 4 ng/l, 8 h after dexamethasone (1.5 mg orally at 2300 h). In a range of pathological conditions ACTH concentrations accurately reflect the disorders of the pituitary-adrenal axis. Endogenous ACTH immunoactivity is stable in vitro at 22 degrees C for at least 1 h in whole blood and at least 4 h in plasma. It is concluded that this two-site IRMA for ACTH in unextracted plasma offers a reliable assay for clinical purposes. PMID- 3026692 TI - Re-evaluation of the clinical value of the 30 min ACTH test in assessing the hypothalamic-pituitary-adrenocortical function. AB - The insulin hypoglycaemia test has been widely used for assessing the hypothalamic-pituitary-adrenocortical function. The outcome of this test has been compared to that of the 30 min ACTH test in 200 consecutive patients with proven or suspected hypothalamic-pituitary disorder. A significant correlation (R = 0.83, P less than 0.001) between the results of the two tests was found. In patients in whom blood glucose levels fell to 2.2 mmol/l (40 mg/dl) or below, the ratio between the peak plasma cortisol concentration during hypoglycaemia and plasma cortisol concentration 30 min after ACTH was 1.002. In patients in whom blood glucose concentration declined to 2.3-3.0 mmol/l (41-54 mg/dl) the ratio was significantly lower (0.828). In eight patients discordant results between the two tests were found. In two patients the ACTH test was normal despite impaired response to insulin. They were studied shortly after acute symptoms of a pituitary adenoma necrosis. Two patients had subnormal responses to insulin despite the fact that pituitary-adrenal function appeared to be intact. In both cases relative insufficiency of the hypoglycaemic stimulus was the likely cause of the discrepancy. In the remaining four patients results of one test was marginally below, and that of the other test marginally above the lower limit of normal. It is concluded that the 30 min ACTH test is reliable for assessing integrated hypothalamic-pituitary-adrenal function (except shortly after acute ACTH deprivation). PMID- 3026693 TI - Escape from dexamethasone-induced ACTH and cortisol suppression by corticotrophin releasing hormone: modulatory effect of basal dexamethasone levels. AB - The response of ACTH and cortisol to corticotrophin-releasing hormone (CRH) after pretreatment with various doses of dexamethasone was investigated in five healthy subjects. The five subjects participated in six experiments. In each experiment 200 micrograms ovine CRH was administered as an i.v. bolus injection at 0900 h after pretreatment with respectively: (A) 1 mg dexamethasone orally at 2300 h in the evening before CRH injection, (B) 2 mg dexamethasone orally at 2300 h in the evening before CRH injection, (C) 4 mg dexamethasone orally at 2300 h in the evening before CRH injection, (D) 2 mg dexamethasone orally at 2300 h in the evening before CRH injection, followed by 2 mg dexamethasone orally 1 h before CRH, (E) no dexamethasone and (F) 1 mg dexamethasone orally 1 h before CRH injection. In spite of overnight suppression with a single dose of dexamethasone CRH elicited cortisol rises in all individuals (experiments A-C). Dexamethasone pretreatment in experiment D abolished the CRH-induced stimulation of the pituitary-adrenal axis. There was a significant and negative correlation between the basal dexamethasone levels (i.e. the dexamethasone levels immediately before CRH administration) in the experiments A-D and the areas under the individual ACTH (R = -0.62; P less than 0.01 by Spearman's rank correlation test) and cortisol (R = -0.81; P less than 0.001 by Spearman's test) curves, i.e. the lower the basal dexamethasone levels, the greater the rise in ACTH and cortisol levels after CRH administration.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3026694 TI - Extramammary Paget's disease. PMID- 3026695 TI - Uptake of free adenosine and adenosine from adenosine monophosphate by human peripheral blood lymphocytes: possible kinetic role for ecto-5'-nucleotidase in the regulation of intracellular adenosine. AB - The role of 5'-nucleotidase in the uptake of adenosine from AMP was investigated in lymphocytes from normal subjects and patients with common variable hypogammaglobulinaemia (CVH) and chronic lymphatic leukaemia (CLL). At physiological pH, the Km values for the uptake of adenosine and of adenosine from AMP by intact cells were one order of magnitude higher than the Km values for 5' nucleotidase. The Vmax values for the hydrolysis of AMP by 5'-nucleotidase were two orders of magnitude greater than for the uptake of adenosine itself or the uptake of adenosine from AMP by normal lymphocytes. 5'-Nucleotidase activity is clearly not rate-limiting in normal lymphocytes for uptake of adenosine from AMP in steady state conditions. Patients with common variable hypogammaglobulinaemia showed a low Vmax for 5'-nucleotidase assayed at pH 7.4 in intact cells as compared to values from control subjects. Michaelis constants (Km) for the uptake of free adenosine and adenosine from AMP as well as 5'-nucleotidase were similar compared to those obtained for controls. The uptake of adenosine moiety from AMP in CLL lymphocytes with a low Vmax for 5'-nucleotidase was also reduced, although not to the same extent as the reduction in 5'-nucleotidase activity. One CLL patient with supranormal levels of 5'-nucleotidase activity showed elevated uptake of adenosine moiety from AMP and of free adenosine. These results suggest that 5'-nucleotidase can influence the salvage of purine by lymphocytes from extracellular nucleotides but only when the enzyme activity is greatly reduced. PMID- 3026696 TI - A canine model of induced purine nucleoside phosphorylase deficiency. AB - Purine nucleoside phosphorylase (NP EC 2.4.2.1) deficiency in man is associated with selective T cell dysfunction and normal B cell immunity. To create an in vivo model of this immune deficiency, we administered 8-aminoguanosine to dogs. This water soluble nucleoside was rapidly converted by NP to the more potent product inhibitor 8-aminoguanine, which had a Ki of 0.52 microM. The accumulation of inosine and exogenous deoxyguanosine in plasma provided evidence that administration of 8-aminoguanosine was effectively inhibiting NP activity. Four dogs given 8-aminoguanosine and deoxyguanosine concurrently for 5 consecutive days showed mean reductions in peripheral blood lymphocytes of 65 +/- 9% range (55-75%) over the test period. Granulocytes, red blood cells, and plateletes remained within the normal range. Administration of 8-aminoguanosine to dogs provides a model of NP deficiency that will permit studies of the specific control of lymphopoiesis and in-vivo immune function. PMID- 3026697 TI - Evidence for disregulation in the control of Epstein-Barr virus latency in patients with AIDS-related complex. AB - Patients affected by AIDS-related complex (ARC) show several immunological abnormalities which may lead to a disregulation of immunosurveillance against viral latent infections. In this paper evidence for Epstein-Barr virus (EBV) reactivation in seven out of eight affected by persistent generalized lymphadenopathy is given. These patients showed either elevated levels of circulating EBNA-positive transformed cells or depressed EBV-specific T cell cytotoxicity, as assessed by the regression assay, or both. A direct involvement of EBV in the pathogenesis of ARC is thus suggested. Natural killer cell activity was found decreased, correlating to the evidence of circulating EBV-infected cells and of impaired EBV-specific immune control. These data provide evidence that, when specific immune mechanisms lose control on ubiquitous latent viruses, the risk for reactivation becomes higher. In the case of EBV, direct evidence of this event is provided by the emergence in the peripheral blood of clones of infected cells with unlimited growth potential. PMID- 3026698 TI - Extraction of immune and inflammatory cells from human lung parenchyma: evaluation of an enzymatic digestion procedure. AB - The inflammatory and immune cell populations of the human lung parenchyma have not been characterized in detail. This report describes a novel and efficient procedure for their extraction. Histologically normal human lung tissue samples from pneumonectomy specimens were sliced to 0.5 mm, and digested in collagenase/DNAse. Viable mononuclear cell yields ranged from 15-48 X 10(6)/g, and were markedly in excess of reported methods employing mechanical tissue disruption, which normally yield populations containing almost exclusively macrophages. The lung digest population was examined by flow cytometry using monoclonal antibodies against cell surface receptors, and found to comprise up to 40% T lymphocytes, 10% B lymphocytes and 30% macrophages, contaminated by less than 1% peripheral blood cells. Based upon these figures, the recoverable lung parenchymal lymphoid cell pool appears considerably larger than previously recognized, being of the same order as the peripheral blood pool. Initial functional studies suggest that such cellular activities as antigen-specific T cell proliferation, antigen-presentation, interleukin 1 production and natural killer cell activity survive the extraction process, and controlled enzymatic digestion experiments with peripheral blood cells indicate that the degree of enzyme-mediated damage to these functions and to cell-surface structures, was minimal. The extraction method thus appears suitable for studying the types and functions of human parenchymal lung cells in health and disease. PMID- 3026700 TI - Inhibition of virus replication does not alter malignant rabbit fibroma virus induced immunosuppression. AB - Malignant rabbit fibroma virus (MV) directly suppresses generation of antibody responses and mitogen induced T and B lymphocyte proliferation. We investigated whether this phenomenon required expression of the complete viral genome. Phosphonoacetic acid (PAA) inhibits poxvirus specific DNA polymerases. Adding PAA to cultures reduces both MV replication and mitogen-driven rabbit lymphocyte proliferation in a dose-dependent fashion. A dose of PAA adequate to inhibit MV replication by about 97%, but insufficient to reduce lymphocyte proliferation appreciably, does not affect the ability of MV to suppress lymphocyte proliferation or initiation of antibody production. Spleen cells from MV tumour bearing rabbits contain very little virus, but inhibit the proliferative and antibody forming responses of normal spleen cells. This activity is shown here to reflect the production by T lymphocytes of a soluble mediator of greater than 25 kD molecular weight. Adding PAA to these mixed spleen cell cultures does not alter the ability of MV to induce T suppressor activity in host lymphocytes. Thus, these immunosuppressive capabilities of MV appear to reflect early MV gene functions. PMID- 3026699 TI - Immunoregulation of lung fibroblast growth: alteration in asbestos-induced pulmonary fibrosis. AB - Initial studies on mononuclear cell-fibroblast interactions have shown that stimulated human lymphocytes produced a fibroblast growth inhibitory factor and that asbestos, a fibrogenic dust, interferes with this process in vitro. To investigate the role of these interactions in pathologies characterized by pulmonary fibrosis, we used a rat model of asbestos-induced fibrosis. Rats received a single intratracheal instillation of either saline or 10 mg of chrysotile asbestos fibres. Three months after treatment, peripheral blood mononuclear leucocytes (PBML) supernatant fractions were prepared and their effects on lung fibroblast growth measured. As for human PBML, rat PBML stimulated with Concanavalin A (Con A) produced 24 h after initiation of the cultures a soluble factor which inhibits lung fibroblast DNA synthesis and growth in a dose-dependent fashion. By contrast, Con A-stimulated PBML from rats exposed to asbestos failed to produce significant levels of fibroblast growth inhibitory activity. No significant change of total PBML number or in the proportion of circulating mononuclear cell populations was observed. Furthermore, upon stimulation with lipopolysaccharide (LPS), monocytes from asbestotic animals retained their capacity to produce interleukin (IL-1), a mediator required for lymphokine production. Our study demonstrates that suppression of FGIF production by circulating PBML occurs in animals with lung fibrosis and suggests that mechanisms other than impairment of IL-1 production may be responsible for the suppressive effect of asbestos on the production of such fibroblast regulatory lymphokine. PMID- 3026702 TI - Naloxone attenuates augmentation of cAMP levels and arrhythmias following myocardial ischaemia and reperfusion in the isolated perfused rat heart. AB - The effects of myocardial ischaemia and reperfusion on arrhythmias and cAMP levels were studied using the Langendoff isolated perfused rat heart preparation. Myocardial ischaemia and reperfusion caused arrhythmias and augmentation of cAMP levels concurrently, supporting the suggestion that myocardial cAMP is related to arrhythmias. Pretreatment with naloxone attenuated both arrhythmias and augmentation of cAMP levels to a similar extent. The results suggest that the anti-arrhythmic effect of naloxone may involve myocardial cAMP. PMID- 3026701 TI - Changes in oxidative burst capacity during murine malaria and the effect of vaccination. AB - Adherent spleen and liver cells from mice infected with Plasmodium yoelii 17X or P. chabaudi AS were tested for production of reactive oxygen intermediates to measure their state of activation. Phorbol myristate acetate (PMA) was used to trigger the respiratory burst and production of superoxide anions was measured by the reduction of nitroblue tetrazolium. Spleen cells from mice infected with P. chabaudi showed an early increase in oxidative activity on day 3, and when the oxidative capacity of the whole spleen was calculated, it was maximal on day 9, just as the mice began to recover. In mice infected with P. yoelii, spleen cells showed an early peak in activity on day 5, and then returned to normal, although the mice did not recover for a further 2-3 weeks. However the total oxidative capacity of the spleen remained high throughout the infection. Mice vaccinated against P. yoelii with a killed blood-stage vaccine showed increased activity on day 3 (spleen) and day 5 (liver), compared with infected control mice. Thus macrophages in these organs could, if given an appropriate trigger, release high levels of these potentially toxic molecules during infection. PMID- 3026703 TI - Characterization of angiotensin converting enzyme from rat tissue by radio inhibitor binding studies. AB - Angiotensin converting enzyme (ACE) derived from rat lung, aorta, epididymus, brain, kidney and plasma was characterized by radio-inhibitor (125I-MK351A) binding studies. Under optimal binding conditions at equilibrium 125I-MK351A bound to ACE was displaced from ACE in a concentration related manner by unlabelled MK351A. MK351A binding site concentration for each tissue and equilibrium dissociation constant (KD) was estimated by Scatchard analysis of binding data. Binding sites/mg protein was greatest in lung and least in brain. The KD for kidney ACE was significantly higher than that of lung, aorta, epididymus or brain ACE (P less than 0.005; t-test, d.f. = 10). 125I-MK351A bound to ACE prepared from lung and kidney was displaced in a concentration dependent manner by SQ20881, SQ14225, MK422, and Ro31-3113-000. Concentration of ACE inhibitor required to displace 50% of bound 125I-MK351A (DD50) was consistently higher for kidney-derived ACE than lung-derived ACE. The differences in radio inhibitor binding characteristics of ACE from different rat tissues suggests that the enzyme active site may not be identical in all organs. PMID- 3026704 TI - Quantitation of (C1INH)2 C1r-C1s complexes in glomerulonephritis as an indicator of C1 activation. AB - C1 activation was assessed in several forms of glomerulonephritis by radioimmunoassay quantitation of circulating (C1INH)2 C1r-C1s complexes (INC). Eight patients with active systemic lupus erythematosus (SLE) and nephritis had elevated serum INC (mean = 15.3 vs control = 5.8, P less than 0.01). Their INC levels were normal during remission. Serum INC had a weak inverse correlation with serum C1q greater than 3 mg/dl (r = 0.42, P = 0.02). In longitudinal studies, serum INC also had a weak inverse correlation with serum C3 and C4. Only 1 of 10 patients with type I and 1 of 15 with type III membrano-proliferative glomerulonephritis (MPGN) had elevated serum INC. No patient with type II MPGN had elevated levels. Two of 10 patients with poststreptococcal glomerulonephritis (P-SGN) had elevated serum INC, but all normalized with convalescence. Patients with IgA nephropathy had normal serum INC. The data demonstrate the importance of C1 activation in SLE and P-SGN. The mechanism of complement activation in types I and III MPGN remains unclear; the data suggest, but do not prove, that C1 independent complement activation may occur in these patients. PMID- 3026705 TI - Typing of intraglomerular mononuclear cells associated with transplant glomerular rejection. AB - To investigate the pathogenesis of glomerular injury in renal allografts, we have analyzed intraglomerular mononuclear cells from 20 biopsies with typical features of transplant glomerular rejection (TGR) (segmental or global occlusion of capillaries by swollen cells). Ten biopsies showing cellular rejection but no glomerular pathology were selected as controls. Microwave fixation and an avidin biotin immunoperoxidase technique were used with the following monoclonal antibodies; Leu1 and OKT3 (pan T cell), Leu 3 a + b and OKT4 (helper T cell), OKT8 (cytotoxic T cell), OKB7 (B cell), OKM1 (monocyte) and OKDR (DR positive cell). The results showed a significant increase of T cells, helper T cells, cytotoxic T cells and monocytes in the patients with TGR compared with the controls (all p less than 0.001, Mann-Whitney U test). Of the T cell subsets, cytotoxic T cells outnumbered helper T cells by a mean ratio of 3.2:1. In the interstitium, the distribution of mononuclear cells was not different between the two patient groups. In both, T cells and monocytes were predominant and few B cells were found. The percentage of cytotoxic T cells was similar to that of helper T cells. In this study, there were at least four TGR patients without cytomegalovirus (CMV) infection and the distribution of intraglomerular mononuclear cells in these patients was indistinguishable from that of other TGR patients. There was no significant association of the distribution of mononuclear cells with the severity of glomerular damage. These results suggest that T cells, predominantly of the cytotoxic subset and monocytes are involved in the mediation of TGR.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3026706 TI - Electron microscopy of chronic eosinophilic pneumonia. AB - We have investigated two cases of chronic eosinophilic pneumonia using the electron microscope. The alveolar septa were thickened due to edema and an infiltrate of numerous mononuclear cells and eosinophils, with a few lymphocytes and occasional plasma cells. Macrophages were often located close to eosinophils and extracellular eosinophilic granules. Occasional eosinophilic granules were observed in the cytoplasm of mononuclear cells. The most striking finding was the presence of distinctive elongated, narrow, tubular inclusions in the cytoplasm of several of the mononuclear cells. These inclusions presented complex curved profiles which sometimes terminated in small, dilated, dense vesicles. Some of the narrow tubular sections of the inclusions presented a pentalaminar structure. Elsewhere, the tubular structures showed localized globular dilatations which contained granular material. Elongated strands of electron-dense material, identical to that forming the intracytoplasmic inclusions, were also located extracellularly, between adjacent mononuclear cells and between mononuclear cells and eosinophils. These inclusions are considered to be the product of phagocytosis of cellular debris and to be related to phagolysosomes rather than to Birbeck granules. PMID- 3026707 TI - The molecular basis of atherosclerosis: concepts derived from studies of lipoprotein metabolism and cell biology. PMID- 3026709 TI - The heart as an endocrine gland. AB - Two independent series of biomedical investigations have led to the discovery that the atria constitute a peptide-secreting endocrine gland. The first investigation is mainly morphological and started with the finding that mammalian atrial (but not ventricular) cardiocytes contain "dense bodies." These "dense bodies," later called "specific granules," were found to be different from lysosomes; to be made up of proteins; and to incorporate both 3H-leucine and 3H fucose in a pattern typical of peptide-secreting endocrine cells. The finding that rat atrial granulation varied with the sodium and water balance led to the crucial observation that atrial extracts have natriuretic and diuretic effects. In less than five years, this new natriuretic hormone has been purified, sequenced and synthesized, and its CDNA and gene have been cloned. The atrial natriuretic factor (ANF) gene has been assigned to the distal short arm of chromosome 1 in band 1P36, while the mouse gene is localized in chromosome 4. The native and synthetic hormones exert identical wide ranging effects (possibly through particulate guanylate cyclase stimulation and adenylate cyclase inhibition) on the kidney, blood vessels, adrenal cortex, and pituitary. Physiopathologic implications of the hormone in experimental hypertension, congestive heart failure, and expansion of blood volume are already beginning to emerge. Concurrently, the search for the function of natriuretic hormones or factors (through studies of negative pressure breathing, atrial distension experiments, head-out water immersion, expansion of blood volume, Na+/K+-ATPase inhibition, and parabiosis experiments in Dahl rats) has provided a general framework within which to interpret this new cardiac function. PMID- 3026710 TI - Osteoarthritis associated with a ball-and-socket ankle joint. A case report. AB - A ball-and-socket joint is an unusual disturbance of normal ankle development. Previous reports have noted good function and the absence of osteoarthritic change in ankles with this configuration. A 50-year-old man with no history of injury developed an osteoarthritic ball-and-socket deformity of the ankle joint. Associated musculoskeletal anomalies included toe syndactylization, genu valgum deformity, and tarsal coalition, a condition that may predispose to the development of this ankle anomaly. Associated valgus deformity of the ankle contributed to the osteoarthritic changes. The conspicuous absence of osteoarthritis in other joints of patients previously reported with this abnormality may be explained by the lack of long-term follow-up examinations. PMID- 3026708 TI - The future of hypertension and renin research. PMID- 3026711 TI - Collagenase chemonucleolysis via epidural injection. A review of 252 cases. AB - Since 1975, collagenase has been injected into 252 herniated intervertebral discs for treatment of sciatica. Highly purified preparations of Chinese collagenase were extracted from Clostridium histolyticum CH 1093 in lyophylized form, which had a specific collagenolytic activity. It had a wide margin of safety as proven by repeated experiments in dogs by means of intradiscal, extradural, intradural, and intravenous injections. One hundred to two hundred units of collagenase dissolved in 5 ml of normal saline was injected epidurally in front of the line joining posterior vertebral body margins. The basic assumption is that the injected enzyme digested the nucleus pulposis. Symptoms disappeared or improved gradually within three to eight weeks. The total success rate was about 77%. For recurrent cases of those with less than satisfactory results, reinjections were performed without any visible antigenic reactions or neurologically adverse effects. PMID- 3026712 TI - [Primary systemic amyloidosis with polyneuropathy as the major symptom demonstrating marked myocardial uptake of technetium-99m-hydroxy methylene diphosphonate]. PMID- 3026713 TI - [Recurrent myoclonus associated with Epstein-Barr virus infection in an elderly patient]. PMID- 3026715 TI - [An autopsy case of generalized variant of Pick's disease]. PMID- 3026714 TI - [Dermatomyositis associated with mediastinal germ cell tumor]. PMID- 3026716 TI - Abnormalities of nerve conduction and somatosensory evoked potential in Poland's syndrome. AB - We report a case of Poland's syndrome with associated primary generalized tonic clonic epilepsy. Somatosensory evoked potentials at median nerve stimulation showed delayed latencies and reduced amplitudes of the evoked response from the hypoplastic right upper limb. Nerve conduction studies showed reduced mixed nerve conduction velocity in the right ulnar nerve and decrease in amplitudes of the compound nerve action potentials in the right median and ulnar nerves. We postulate a congenital hypoplasea of the above nerves and brachial plexus in the present case. PMID- 3026717 TI - Angiotensin converting enzyme activity and hypoxia. PMID- 3026718 TI - Studies in vivo and in vitro of terbutaline-induced beta-adrenoceptor desensitization in healthy subjects. AB - Beta-Adrenoceptor function was studied in eight healthy subjects before, during and 24 and 72 h after cessation of 2 weeks continuous oral treatment with the beta 2-adrenoceptor agonist terbutaline (sustained release, 7.5 mg twice daily). In vivo, blood pressure, heart rate, plasma noradrenaline and plasma cyclic AMP responses to isoprenaline (0.01, 0.02 and 0.05 microgram min-1 kg-1 intravenously) were related to the plasma concentrations of isoprenaline. for comparison, beta 2-adrenoceptor function was evaluated in lymphocytes in vitro by studies of isoprenaline-induced accumulation of cyclic AMP and radioligand binding studies using 125I-iodohydroxybenzylpindolol. In vivo, the beta 2 mediated plasma cyclic AMP response to isoprenaline was markedly attenuated during terbutaline treatment and was still reduced by 38% (P less than 0.05) 72 h after discontinuation of treatment. The blood pressure and heart rate responses to isoprenaline were unaffected by treatment. Isoprenaline-induced elevations of plasma noradrenaline concentrations were markedly reduced during terbutaline treatment. This indicates an attenuation of isoprenaline-induced increases in sympathetic nerve function and could explain why no attenuation of the isoprenaline-induced vasodilatation was observed. Thus, plasma cyclic AMP seems to be a better marker than diastolic blood pressure when evaluating beta 2 adrenoceptor responsiveness in vivo in man, since it is not influenced by counter regulatory increases in sympathetic nerve activity and/or noradrenaline overflow from sympathetic nerves. In lymphocytes, the isoprenaline-stimulated cyclic AMP accumulation was reduced by 75% and the beta-adrenoceptor binding sites were reduced by 40% 12 h after dosing. Also the lymphocyte beta 2-adrenoceptors recovered slowly after withdrawal of treatment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3026719 TI - Epidemic polyarthritis and Ross River virus disease. AB - Ross River virus is a mosquito-transmitted alphavirus indigenous to Australia, Papua New Guinea and nearby islands, which recently appeared in other western and central South Pacific islands. Human infection can be manifest by varied constitutional disturbances, rash and rheumatic symptoms, known in Australia as epidemic polyarthritis and broadly similar to certain alphavirus diseases in other regions. Although usually short-lived, the rash can persist for 5 months. Rheumatic effects involve synovial joints, tendon and ligaments, and can continue or recur in peripheral joints and tissues as long as 6 years, though gradually improving without destructive changes. At different times, the disease can closely simulate rubella and other virus diseases, Henoch-Schonlein syndrome, rheumatoid and other chronic rheumatic diseases. Diagnosis rests upon geography, specific serology and judicious interpretation of clinical and supportive laboratory data. Skin and synovial lesions are characterized by infiltration of mononuclear cells. Their pathogenesis most likely depends on the reaction of these cells with persistent foci of virus disseminated during the early viraemic phase of infection. PMID- 3026720 TI - Dietary fiber in disease prevention and treatment. PMID- 3026721 TI - Cytochemical identification of lymphocytes and other mononuclear cells in ovine and bovine hemal nodes. AB - Cytochemical and immunocytochemical studies were carried out with specific markers for B lymphocytes, dendritic reticulum cells (ATPase, 5'Nase, SIg) and T lymphocytes (ANAE, A.P.) in an attempt to identify the mononuclear cells present in bovine and ovine hemal nodes. The results show that primary nodes and mantle of secondary nodes are composed of B cells, whereas T cells are mainly localized in the interfollicular cords. Since such an arrangement resembles the picture in normal lymph nodes, but the direct contact between lymphatic tissue and blood is more reminiscent of the spleen, hemal nodes probably perform immunological functions similar to those of both normal lymph nodes and spleen. PMID- 3026722 TI - [Demonstration of the expression of bovine leukemogenic virus (BLV) in sheep lymphocytes by an immunofluorescence technic using monoclonal antibodies]. AB - An indirect immunofluorescence (IF) test was developed to detect bovine leukemia virus (BLV) antigen expression in infected sheep lymphocytes, using monoclonal antibodies anti BLV-major envelope glycoprotein gp51. Peripheral blood lymphocytes were cultivated for 48 h in presence of phytohemagglutinin (PHA) (50 micrograms/ml), and then fixed with acetone. The cells were assayed for the IF test. All experimentally infected sheep were positive with this test. PMID- 3026723 TI - Immunopharmacology of anaphylatoxin-induced bronchoconstrictor responses. AB - The complement anaphylatoxin peptides, C3a and C5a, are potential mediators of immediate hypersensitivity reactions, eliciting many of the same actions on isolated tissue and cell preparations as specific antigen. Instilled intratracheally in experimental animals, the peptides induce acute bronchospasms and are sometimes lethal. In vitro, they cause dose-dependent contraction of isolated lung tissue preparations, a response which correlates well with bronchospasms observed in vivo, and our current understanding of the cellular and molecular mechanisms of this action are reviewed here. C5a and its catabolic derivative, C5ades Arg, stimulate contraction of isolated guinea pig lung parenchymal strips in part by production of leukotrienes that constitute SRS-A, and by release of histamine. Leukotrienes in turn release thromboxane from lung tissue, and evidence indicates that at least part of the spasmogenic activity of these peptidolipids is mediated by this effect. C3a is considerably less potent than C5a in contracting lung tissues and appears to act primarily by causing the release of spasmogenic cyclooxygenase metabolites. Both peptides may additionally have direct action on contractile cells within the tissue. Platelet-activating factor (PAF), an unusual phospholipid mediator released from inflammatory cells stimulated with C5a and other agents, also contracts isolated lung parenchymal tissues. PAF stimulates release of significant quantities of thromboxane from guinea pig lung; however, indomethacin does not block contractile responses of the tissue. Recent evidence indicates that PAF may act on parasympathetic neurons in lung to release endogenous acetylcholine, and this action may be a major component of tissue responses to this mediator. Thus the complement anaphylatoxins stimulate release of many of the same mediators from lung tissues as are released by antigen challenge of sensitized tissue, and may, therefore, play an important role in the pathogenesis of allergic bronchospasms. PMID- 3026724 TI - Anaphylatoxins: possible roles in disease. AB - Anaphylatoxins, in particular C3a and C5a, have various biological activities which suggest a role as mediators of inflammatory reactions: they cause contraction of smooth muscle, histamine release, increase in capillary permeability, adhesion of leukocytes to vascular endothelium, leukocyte chemotaxis, and aggregation of platelets and leukocytes. Most of these effects are supported by the cooperation of other mediators, in particular arachidonic acid derivatives which may be produced by anaphylatoxin-stimulated cells, e.g. leukocytes or endothelium. In vivo effects of the complement peptides depend very much on the site of their generation: intravascular release in the general circulation leads to adverse symptoms such as adult respiratory distress syndrome and shock lung, mainly due to leukocyte activation, aggregation and their accumulation in lung vessels. Intravascular release may be induced by certain drugs, and by contact of blood with the surfaces of bypass or dialysis apparatus. Induction of local inflammatory and defense reactions requires release of anaphylatoxins in tissue spaces. Tissue fluid differs quantitatively from blood plasma in its concentration of complement components. This raises some problems of how efficient concentrations of C3a and C5a can be attained at the site of a lesion to generate a chemotactic gradient capable of attracting blood leukocytes. PMID- 3026725 TI - Recent findings on 1,25(OH)2 vitamin D3 may provide new concepts for understanding the pathogenesis of uremia. PMID- 3026726 TI - Role of hormonal disturbances in uremic growth failure. PMID- 3026727 TI - Erythropoietin production in cultures of rat renal mesangial cells. PMID- 3026728 TI - Impaired regulation of alpha- and beta-adrenoceptor function in chronic renal insufficiency. PMID- 3026729 TI - Parathyroid hormone secretion. Molecular events and regulation. PMID- 3026731 TI - "Scar sign" on computed tomography and sonography in fibrolamellar hepatocellular carcinoma. AB - Demonstration by computed tomography of a central linear or stellate low attenuation region within a focal liver mass has been reported to be suggestive of focal nodular hyperplasia in appropriate clinical settings. We present a case of a focal liver mass with typical central scarring in which the ultimate diagnosis was fibrolamellar hepatocellular carcinoma. PMID- 3026730 TI - Low-protein diet supplemented with essential amino acids and keto analogues. Effects on uremic polyneuropathy and encephalopathy. PMID- 3026733 TI - Cytochrome oxidase activity of Fuchs' endothelial dystrophy. AB - The normal human corneal endothelial monolayer maintains stromal water equilibrium and thus, transparency, by means of a pump-leak mechanism. Water leaks into the stroma through non-tight lateral cell junctional complexes and is drawn out by an energy dependent cell membrane ion pump. We investigated the histochemical localization of cytochrome oxidase activity (CO), an important energy-deriving mitochondrial enzyme in dysfunctional corneas with Fuchs' endothelial dystrophy (ED), which is a regionally distributed disease. Keratoconus corneas were used as controls for functional control endothelium. In the central area of the corneal button, decreased CO activity was demonstrated which correlated clinically with central corneal edema. This reflects decreased metabolic activity and/or decreased numbers of mitochondria in the attenuated dysfunctional cells. In the mid-periphery, CO activity was increased in the cellular rosettes surrounding guttata, which may be related to increased synthesis of abnormal Descemet's membrane and guttata. Peripherally, the large polygonal cells resembled functional endothelium in their morphology and CO activity. We have, therefore, demonstrated regional differences in energy metabolism in endothelium from Fuchs' ED patients which may be related to decreased numbers of mitochondria in the dysfunctional cells, and/or to synthesis of abnormal Descemet's membrane material. PMID- 3026732 TI - Inhibition of Na,K-ATPase by sodium selenite and reversal by glutathione. AB - Sodium selenite has been shown to inhibit Na,K-ATPase. Glutathione, at sufficient excess, is able to prevent or reverse the inhibition. Dithiothreitol can also reverse much of the inhibition, but KCN cannot. Selenomethionine does not inhibit Na,K-ATPase. The interactions of sodium selenite with Na,K-ATPase and glutathione may aid in understanding the early events in selenium cataractogenesis. PMID- 3026734 TI - Inhibition of tonin activity by growth hormone and testosterone depletion in one kidney, one-clip hypertensive rats. PMID- 3026735 TI - Altered DNA methylation patterns in human lung carcinomas. PMID- 3026736 TI - Response of varicella zoster virus and herpes zoster to silver sulfadiazine. AB - The addition of silver sulfadiazine to cultures of varicella zoster virus resulted in inactivation of the viral infectivity. At a concentration of 10 micrograms/ml or higher the virus was inactivated after thirty minutes exposure at 37 degrees C. Forty-two patients with herpes zoster were treated topically with 1 percent silver sulfadiazine cream applied four times a day. All patients experienced complete drying of vesicles, marked reduction erythema and edema, and striking elimination of pain and burning sensation within twenty-four to seventy two hours. The sooner the treatment began after the onset of symptoms, the more dramatic was the response. Postherpetic neuralgia was either mild or did not occur. Signs of local, systemic, or laboratory-documented toxicity were not observed. PMID- 3026737 TI - The cellular homologue of the transforming gene of SKV avian retrovirus maps to human chromosome region 1q22----q24. AB - We report the chromosomal localization of the cellular oncogene SKI, the putative oncogene of the Sloan-Kettering viruses (SKVs), a group of transforming retroviruses that had been isolated from chicken embryo cells infected with the avian leukosis virus tdB77. Southern blot analysis of DNA from mouse X human somatic cell hybrids with the v-SKI probe established synteny with chromosome 1, but excluding the region 1pter----q21. In situ hybridization of the same probe both to human spermatocyte pachytene and lymphocyte metaphase chromosomes enabled precise localization of the gene to the region 1q22----q24, a region that frequently is involved in translocations and other rearrangements in diverse human tumor types. In situ hybridization studies of metaphase spreads from a small noncleaved cell lymphoma that exhibited a t(1;14)(q21;q32) translocation showed that SKI translocates to the der(14) chromosome. Cytogenetic analysis of 65 prospectively ascertained non-Hodgkin's lymphomas revealed that the SKI region undergoes nonrandom breakage leading to translocations. Further analysis of the chromosome breaks in this group of lymphomas suggested that those involving the SKI site probably are of importance in tumor progression. PMID- 3026738 TI - [Metastases of malignant tumours to the parotid glands. Report of 2 cases]. PMID- 3026739 TI - Antiviral therapy and pulmonary disease. PMID- 3026740 TI - Discrepancy between the antibacterial activities and the inhibitory effects on Micrococcus luteus DNA gyrase of 13 quinolones. AB - Thirteen quinolone antibacterial agents were investigated as to their ability to inhibit Micrococcus luteus DNA gyrase and cell growth, and compared to those of novobiocin and coumermycin. Among the quinolones tested, the most active were found to be CI-934 and ciprofloxacin, which inhibited gyrase full supercoiling activity at concentrations of 100 and 200 micrograms/ml, respectively, while inhibiting cell growth at a concentration of 1 microgram/ml. However, both novobiocin and coumermycin inhibited gyrase full supercoiling activity at concentrations of 0.5 and 1.0 microgram/ml, respectively, which were comparable to those concentrations causing inhibition of cell growth. PMID- 3026741 TI - Detection of human papillomavirus in epithelial cells of uterine cervix-frequency of types 16 and 18 HPV. Preliminary results of a clinical cytological and virological study. PMID- 3026742 TI - Na+ + K+ ATPase of erythrocyte membrane in progressive muscular dystrophy. PMID- 3026743 TI - Strain of hepatitis A virus causing cytopathic effects isolated in A549 cell line. PMID- 3026744 TI - Ultrastructural study of primary hepatocellular carcinoma. An approach to grading. PMID- 3026745 TI - BK virus antibody in population of various age groups in Beijing. PMID- 3026746 TI - Detection of herpes simplex virus antigens in tissue cultures by avidin-biotin peroxidase complex method. PMID- 3026748 TI - [Digestive linitis plastica. Apropos of a case of successive gastric and rectal localization]. PMID- 3026747 TI - [2 cases of mucoid pseudocysts of the peripheral nerves]. PMID- 3026749 TI - [Linitis plastica of stomach. Study of 102 cases surgically treated. Results of the surgical treatment]. PMID- 3026750 TI - [Double gastric and colorectal localization of linitis plastica]. PMID- 3026751 TI - [Radiologic manifestations and pathologic basis of the histologic subtypes of small cell carcinoma of the lung--a radiologic-pathologic correlative study of 70 cases]. PMID- 3026753 TI - [The analysis of the doubling time for 115 cases of primary peripheral carcinoma of lung]. PMID- 3026752 TI - [Roentgen diagnosis of senile pulmonary tuberculosis coexisting with primary lung cancer (an analysis of 108 cases)]. PMID- 3026754 TI - [The 60Co radiotherapy of intracranial tumor (an analysis of 170 cases)]. PMID- 3026755 TI - [The study of the relationship between polymerized human serum albumin receptor and HBV replication]. PMID- 3026757 TI - Relationship of backwash ileitis to ileal pouchitis after ileal pouch-anal anastomosis. AB - To assess whether the presence of backwash ileitis predisposed to the subsequent development of ileal pouchitis after ileal pouch-anal anastomosis, 131 patients who had the operation were studied. Fifteen patients had nonspecific inflammation in the terminal ileum noted at the time of the operation, while 20 patients subsequently developed pouchitis. No correlation between the two conditions was found. Pouchitis developed in two of 15 patients (13 percent) with backwash ileitis and in 18 of 116 patients (16 percent) without the ileitis (P greater than 0.05). Among the 20 patients with pouchitis only two (10 percent) had had previous backwash ileitis. It is concluded that the presence of backwash ileitis does not predispose to later development of pouchitis, and so does not contraindicate use of the inflamed terminal ileum for construction of the ileal pouch and anastomosis. PMID- 3026756 TI - Comparison of tissue disaggregation techniques of transitional cell bladder carcinomas for flow cytometry and chromosomal analysis. AB - DNA index (DI) measurements and chromosomal analysis of 42 transitional cell carcinomas were done after mechanical and enzymatical disaggregation of the tumor specimens. The results obtained with these different disaggregation techniques were compared in the 33 cases (79%) that showed recognizable chromosomes. The enzymatically obtained cell suspensions could not be used for chromosomal analysis after short-term culture of 24 hours. In four cases, the DI after enzymatical treatment could not be estimated. In most cases, the DI obtained from the tumor cells was similar for both aggregation techniques, with the exception of four cases of enzymatically treated cell suspensions in which the DI could not be estimated. The average DI of the aneuploid tumors was 13% higher than the corresponding chromosome count. In 19% of the aneuploid tumors the proportion of aneuploid cells could not be measured after enzymatical treatment. In the remaining suspensions the proportion of diploid cells was higher after enzymatical disaggregation than after mechanical treatment. It is concluded that for flow cytometric and direct chromosomal analysis of bladder tumors, the mechanical disaggregation technique is most suitable. PMID- 3026758 TI - Bowenoid papulosis. AB - Bowenoid papulosis is a recently described condition with a clinically benign appearance of multiple reddish-brown or violaceous papules. The histologic picture, however, is that of squamous-cell carcinoma in situ. In the reported cases the lesions usually occurred on the genitalia of patients in the third or fourth decades of life. We recently encountered a patient who presented with papules in the anal area. Human papillomavirus has been identified as the causative agent. Differentiation from Bowen's disease is usually possible due to the presence of multiple small lesions and the young age of the patients. The long-term consequences of bowenoid papulosis are still unknown. The presence of human papillomavirus in a female or her sexual partner places her at higher risk for cervical neoplasia. The treatment of bowenoid papulosis is ablation or 5-FU cream. PMID- 3026761 TI - [Plasmid vector for gene expression containing the temperature-dependent promoter of phage lambda P'R DNA]. PMID- 3026760 TI - Overwhelming pneumonia. AB - Overwhelming pneumonias remain an important cause of morbidity and mortality. These illnesses may be rapidly fatal; thus, many patients are treated empirically. Although the various etiologic agents cannot be differentiated on the basis of radiographic appearance, epidemiologic information may give a clue to the cause. Community-acquired overwhelming pneumonias are usually due to pyogenic bacteria (especially Streptococcus pneumoniae), mycoplasma, mycobacteria, and fungi. Hospital-acquired pneumonias are usually due to aerobic gram-negative bacilli. If the patient is immunocompromised, Pneumocystis carinii, Candida, and Aspergillus must be considered. Choice of optimal antimicrobial therapy requires that a specific etiology be identified. Gram's stain of sputum is often helpful in the diagnosis of community-acquired pneumonia. Invasive diagnostic techniques such as bronchoscopy and open lung biopsy are often required in nosocomial pneumonias and pneumonias in immunocompromised patients. PMID- 3026759 TI - Sequential morphologic and biochemical studies of naturally occurring wheat sensitive enteropathy in Irish setter dogs. AB - This study has investigated the potential role of wheat in the pathogenesis of a naturally occurring enteropathy in Irish setter dogs. At eight months on a cereal containing diet, jejunal biopsies from affected animals exhibited partial villus atrophy, increased intraepithelial lymphocytes, and distinct biochemical abnormalities in the brush border. Activities of alkaline phosphatase and leucyl 2-naphthylamidase were almost undetectable while disaccharidases were unaltered. Activity of 5'-nucleotidase (basolateral membrane) was low, and reduced malate dehydrogenase reflected a loss of mitochondrial activity, but other organelles were unaffected. Recovery was achieved on a wheat-free diet. Relapse on subsequent wheat challenge was characterized by partial villus atrophy and a selective effect on the brush border: modal density was decreased and there was a severe loss of brush-border alkaline phosphatase activity. These findings document a wheat-sensitive enteropathy in Irish setter dogs and suggest that brush-border alkaline phosphatase is specifically susceptible to damage by wheat. PMID- 3026762 TI - [Isolation of consistently unstable strains of Drosophila melanogaster]. PMID- 3026763 TI - [Point mutation clusters predetermined by the quasi-palindromic nucleotide sequence in DNA]. PMID- 3026764 TI - [Intrauterine fetal death following erythema infectiosum]. PMID- 3026765 TI - [Cytomegaly infection in infancy in a family with demonstrated HIV antibodies]. AB - Cytomegalovirus (CMV) was isolated from the urine of babies from two consecutive pregnancies of a mother. The first baby had died of pneumonia at the age of six months, two years before the second confinement (twins of different sexes). The male twin was examined at the age of eight months on account of generalized lymphomas, pulmonary infiltrates and thrombopenia. He excreted CMV in the urine, and HIV antibodies were found in the serum. All members of the family (also the baby who died in 1983, from whom serum samples had been stored at -20 degrees C) finally proved to be carriers of antibodies against HIV. The parents and the female twin did not show any clinical symptoms indicating a HIV infection; however, CMV was also isolated from a urine sample from the mother. PMID- 3026766 TI - [Porphyria cutanea tarda: pathognomonic needle-like inclusions in the cytoplasm of the hepatocytes]. PMID- 3026767 TI - [Transient HIV-antibody positive tests in heart transplantation patients after repeated administration of a CMV immune globulin preparation]. AB - After repeated administration of relatively high doses of a cytomegalovirus immunoglobulin preparation to two patients after heart transplantation, both the ELISA and the immuno-blot tests became temporarily positive (for 21 and 41 days, respectively) to HIV antibodies. The CMV immunoglobulin preparation was demonstrated to have high HIV antibody titres. The latter ran parallel to antibody titres against measles virus, also transferred passively with the immunoglobulin preparation. This makes it unlikely that HIV antibodies had been formed by active immunization from HIV in the immunoglobulin preparation. Within 126 and 176 days, respectively, after the last positive test against HIV antibodies there was no further demonstration of HIV antibodies or of a corresponding illness. There was thus no evidence of an HIV infection having been transmitted by the immunoglobulin preparation. PMID- 3026769 TI - [Virus-related gastroenteritis]. PMID- 3026770 TI - [Immunogenicity of inactivated, multivalent IB/ND oil emulsion vaccine in hybrid laying and fattening animal flocks]. PMID- 3026768 TI - [Cytomegalovirus-associated superinfection of the lung following kidney transplantation]. AB - In a kidney-transplant patient, there was superinfection of the lungs by Klebsiella pneumoniae and Legionella pneumophilia with multiple necroses in the course of a primary cytomegalovirus infection. In a later phase of the disease, there was an opportunistic colonization of the necrotic cavities and the adjacent lung tissue by Aspergillus fumigatus and Candida albicans. The cytomegalovirus infection led to a pronounced cellular immunosuppression in the patient. This favored superinfections by bacteria and fungi. A poor nutritional state and a chronic lung disease are to be considered as predisposing risk factors. PMID- 3026771 TI - [The clinical use of angiotensin converting enzyme inhibitors]. PMID- 3026772 TI - [Adenomas and carcinomas of the small intestine in familial polyposis and Gardner syndrome--analysis of a literature survey]. AB - An analysis of 70 case records from the literature of patients with familial polyposis or Gardner syndrome and adenomas or carcinomas of the small bowel demonstrated that there is no difference between the neoplasms of the small bowel in familial polyposis or in Gardner syndrome. Between the adenomas and the carcinomas of the small bowel it was possible to show, that there exists a similar close relation as in the large intestine. In some carcinomas of the small bowel rests of adenomas can be observed. The peak incidence of small bowel adenomas is more than a decade earlier than that of carcinomas. The distribution of adenomas in the small bowel is quite similar to that of carcinomas. Therefore it can be supposed, that the significance of familial polyposis coli and Gardner syndrome for the importance of an adenoma-carcinoma-sequence in the large bowel is also existing in the small bowel. PMID- 3026773 TI - [Monoclonal antibodies to RPMI-1788 lymphoblastoid line cells]. AB - Monoclonal antibodies IPO-1--IPO-8 against surface antigens of B-lymphoblastoid cell line RPMI-1788 were prepared. Murine hybridomas were obtained by fusion of immune spleen cells and myeloma cells. Screening of specific antibody production was carried out by indirect immunofluorescence. Expression of these antibodies against a panel of 11 human cell lines was carried out by indirect immunofluorescence. Expression of antigens detected with IPO-1--IPO-8 were investigated on peripheral blood cells of healthy individuals and patients with CLL, ALL, myeloma and on lymph node cells of patients with Hodgkin's disease. Specificity of these MoAbs is discussed. IPO-5 is shown to react with the HLA related determinant. The antibody IPO-3 appears to recognize a differentiation antigen of human B cells. PMID- 3026774 TI - [Motor conduction time of the corticospinal tract in spinal stimulation]. AB - A muscular action potential (MAP) following transcranial and spinal stimulation can be recorded from most of the striped muscles. The motor central conduction time after transcranial and spinal stimulation can be determined at cervical 6/7. It is at 5.0 ms in normal subjects. This was demonstrated in four normal subjects. After spinal stimulation at cervical 6/7 a motor evoked potential can also be recorded from the scalp. It has a latency that is comparable to motor conduction time. In animal experiments in examining ten rabbits a motor evoked potential to stimulation at several levels of the spinal cord could also be determined. PMID- 3026775 TI - [Changes in brain stem potentials in dystrophia myotonica]. AB - In 15 patients with dystrophia myotonica brainstem auditory potentials (BAEP) were examined: in 8 patients (53%) pathological components in the BAEP's (such as increased latency of one peak) and in 80% a pathologic component in the neurography could be found. Comparing the latencies of the peak and the interwave latencies in the patients' and control group there was no significant difference. Hearing disturbances influenced the latencies of the first BAEP-component, increased triglycerides correlated with pathologic nerve conduction velocity and singular pathologic BAEP values correlated with abnormal neurographic parameters. All these correlations emphasize the complexity of alterations in dystrophia myotonica. PMID- 3026776 TI - Use of the reverse hemolytic plaque assay to study the regulation of anterior lobe adrenocorticotropin (ACTH) secretion by ACTH-releasing factor, arginine vasopressin, angiotensin II, and glucocorticoids. AB - A reverse hemolytic plaque assay (RHPA) for ACTH was developed to study the responses of anterior lobe corticotropes to secretagogues-CRF, arginine vasopressin (AVP), angiotensin II (A-II), or to a 6- to 24-h pretreatment with corticosterone. Tests showed that optimal plaque formation was obtained after 3-4 h with 1:100-1:200 anti ACTH-(25-39) and 1:20-1:50 complement. Under optimal basal conditions, 6.6% of pituitary cells from normal male rats formed plaques. The addition of 50-90 micrograms/ml ACTH to the anti-ACTH for 48 h before its use in the RHPA resulted in a decrease in percentage of ACTH plaques to levels not different from those obtained with preimmune serum, or when complement was omitted from the assay (0.9-1.3%). There was a gradual increase in percentage of ACTH plaques to 9.8% of the population after exposure to increasing doses of CRF (0.1-10 nM). These same high percentages of ACTH plaques could also be obtained by the addition of 1 nM AVP or A-II with the lowest doses of CRF (100-500 pM). Exposure to 1 nM AVP alone resulted in no significant increases in percentages above basal values. Average plaque areas were increased to maximal levels of three to four X basal with increasing doses of CRF. Finally, when cells were pretreated with 100 nM corticosterone for up to 24 h, the percentage of plaques formed under basal conditions was reduced by 60% (2.6%) and it did not increase after exposure to 1 nM CRF. The data from the RHPA correlate well with previous studies of corticotropes. Since ACTH cells normally represent 10% of the anterior lobe cell population, the RHPA shows that a subset of corticotropes (6.6%) is actively secreting under basal conditions tested in this study. The remaining 3% can be stimulated to form plaques by high doses of CRF (greater than 1 nM) or low doses of CRF (100 pM) and 1 nM AVP or A-II. Glucocorticoids reduce the percentage of ACTH plaques formed under basal or stimulated conditions and allow no CRF stimulated increase in plaque area. This correlates with recent reports that show glucocorticoids inhibit proopiomelanocortin messenger RNA synthesis and lower the number of pituitary CRF receptors. PMID- 3026777 TI - Parathyroid hormone-like adenylate cyclase-stimulating activity from a human carcinoma is associated with bone-resorbing activity. AB - We found previously that a human renal carcinoma cell line derived from a hypercalcemic patient induces humoral hypercalcemia when grown as allografts in the nude mouse and secretes a protein that activates adenylate cyclase via the PTH receptor. The purpose of this study was to examine the conditioned medium of this cell line for bone-resorbing activity in vitro. Processed conditioned medium produced dose-dependent stimulation of bone resorption in cultured fetal rat limb bone explants. Two PTH antagonists were used to assess the PTH receptor dependence of this bone-resorbing activity. Neither [8Nle,18Nle,34Tyr]bovine (b) PTH-(3-34) amide nor [34Tyr]bPTH-(7-34)amide inhibited bone resorption or limb bone cAMP accumulation induced by either processed conditioned medium or equivalent concentrations of bPTH-(1-34). As an alternate means to assess whether this tumor-derived PTH-like protein had intrinsic bone-resorbing activity, the latter was measured during partial purification of PTH-like adenylate cyclase stimulating activity (ACSA) from conditioned medium by consecutive gel filtration and reverse phase HPLC. The bone-resorbing activity in conditioned medium could not be resolved from PTH-like ACSA by these two separation techniques, indicating that the activities may be intrinsic to the same protein. These results are consistent with the view that a tumor-derived protein with PTH-like ACSA and bone resorbing activity may be responsible for hypercalcemia in vivo. PMID- 3026778 TI - The effect of transforming growth factor-beta on follicle-stimulating hormone induced differentiation of cultured rat granulosa cells. AB - Growth factors have been shown to modulate differentiation of cultured ovarian granulosa cells. Transforming growth factors (TGFs) constitute a family of polypeptide growth factors capable of reversibly inducing anchorage-independent growth in normal cells. Epidermal growth factor (EGF), which has significant structural homology with TGF alpha, has been shown to modulate differentiation of granulosa cells in vitro. Similarly, TGF beta (TGFB) has been found to have significant structural homology with ovarian follicular fluid inhibin. To examine whether TGFB might affect granulosa cell growth or differentiation, rat granulosa cells were cultured in serum-free medium containing insulin for up to 3 days with varying concentrations of TGFB in the presence or absence of FSH. TGFB caused a dose-dependent increase in FSH-stimulated LH/hCG receptor binding, but had no effect on binding in the absence of FSH; TGFB (10.0 ng/ml) further increased FSH stimulated LH/hCG receptor binding by 48 +/- 8% (P less than 0.02). Similarly, FSH-stimulated progesterone production was increased by TGFB in a dose-dependent manner; TGFB (1.0-10.0 ng/ml) increased FSH-stimulated progesterone production 2- to 3-fold (P less than 0.02). In contrast, EGF (10.0 ng/ml) decreased FSH stimulated LH/hCG receptor binding by 93 +/- 1% (P less than 0.02). Neither FSH stimulated intracellular nor extracellular cAMP accumulations were affected by TGFB treatment. However, EGF (10.0 ng/ml) diminished extracellular and intracellular FSH-stimulated cAMP accumulation at 48 and 72 h of culture. Culture protein and DNA content were not significantly affected by TGFB. These results suggest that TGFB may enhance FSH-stimulated LH receptor induction and steroidogenesis by mechanisms that do not further increase net cellular cAMP accumulation; TGFB and EGF can have opposite effects on gonadotropin-dependent differentiation; and products of the TGFB/inhibin gene family may have a capacity for autocrine or paracrine modulation of granulosa cell differentiation. PMID- 3026780 TI - Lipoprotein requirements for secretion of ultraviolet-absorbing corticosteroids by guinea pig adrenocortical cells in vitro: inner versus outer cortices; zona glomerulosa versus zona fasciculata. AB - Most studies of lipoprotein requirements for steroid secretion by the adrenal have examined the mixed cell population of the whole gland; none have examined lipoprotein requirements of guinea pig adrenocortical cells. In this study the effect of exogenous lipoprotein on the ability of cells from each of the different regions of the guinea pig adrenal cortex to synthesize and secrete steroids has been analyzed in vitro, under baseline and ACTH-stimulated conditions. Most studies have assessed the effects of lipoprotein on one or a few selected steroids. In this study the effects of lipoprotein and ACTH were examined both by an assay for fluorogenic steroids and by HPLC analysis of the spectrum of UV-absorbing steroids. Guinea pig outer adrenocortices, containing zona glomerulosa and zona fasciculata, maintained in vitro as fragments or as isolated cells, secreted at least 5 times more steroid than the inner cortex, predominantly zona reticularis, and were dependent upon lipoproteins for their secretion. Corticosterone (B) and cortisol (F) were the predominant products of both zones. Aldosterone (Aldo), 18-hydroxycorticosterone (18-OH B), deoxycorticosterone (DOC), 11 beta-hydroxyandrostenedione (11 beta-OH And), androstenedione (And), and deoxycortisol were less abundant products of the outer cortex, while the inner cortex secreted only very small amounts of these steroids. Each of the outer cortical cell types secreted a distinct spectrum of steroids. Aldo, 18-OH B, and DOC were characteristic of glomerulosa cells, but B was most prominent. Fasciculata cells secreted primarily F, with 11 beta-OH And as their next most prominent product. Low density lipoprotein (LDL) enhanced steroid secretion by glomerulosa cells to a greater extent than that by fasciculata cells, but the stimulation of LDL utilization by ACTH was greatest for fasciculata cells. LDL and ACTH also influenced the pattern of steroids secreted by each cell type. Addition of LDL to glomerulosa cells enhanced secretion of DOC and B, but not that of Aldo or 18-OH B. In fasciculata cell cultures, LDL enhanced secretion of both F and 11 beta-OH And. ACTH, particularly in the presence of LDL, stimulated secretion by glomerulosa cells of Aldo and 18 OH B, as well as that of F, And, and 11 beta-OH And. The combined presence of ACTH and LDL in fasciculata cell cultures preferentially stimulated secretion of F and B.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3026779 TI - Characterization of gamma-aminobutyric acid receptors in the neurointermediate lobe of the amphibian Xenopus laevis. AB - The neurotransmitter gamma-aminobutyric acid (GABA) is involved in the regulation of secretion of MSH from the intermediate lobe of Xenopus laevis. The purpose of this study was to identify the GABA receptor(s) involved by determination of the effect of specific receptor agonists and antagonists on the release of immunoreactive MSH from superfused neurointermediate lobes of Xenopus. Exogenous GABA induces a rapid inhibition of MSH secretion. There was no evidence for a transitory stimulatory effect of GABA as reported for the rat melanotropes. Both the GABA agonists (GABAa) homotaurine and isoguvacine and the GABA agonist (GABAb) baclofen inhibited MSH release in a dose-dependent manner. In vivo, homotaurine and baclofen caused aggregation of pigment in dermal melanophores. The MSH release-inhibiting effect of homotaurine and isoguvacine could be antagonized by the specific GABAa receptor antagonist bicuculline. However, bicuculline and picrotoxin failed to block the effect of exogenous GABA. We conclude that in the neurointermediate lobe of Xenopus laevis both GABAa and GABAb receptors are present, suggesting a dual inhibitory regulation. PMID- 3026781 TI - The influence of plasma membrane cholesterol on the response of adrenal cells to adrenocorticotropin. AB - The concentration of cholesterol in plasma membranes of Y-1 cells was altered between values of the molar ratio of cholesterol to phospholipids (C/P) of 0.6:1.6 to study the influence of plasma membrane cholesterol on the response to ACTH. Increasing concentrations of membrane cholesterol (C/P) were associated with an increase in the number of ACTH receptors and, hence, production of cAMP and steroids without affecting the nature of binding (Kd) of ACTH to its receptor. Binding studies revealed spare ACTH receptors at all concentrations of membrane cholesterol. However, production of cAMP and steroids by Y-1 cells were tightly coupled. Studies with cholera toxin showed that extreme changes in C/P were without influence on the activity of Ns or that of the cyclase itself. It was also shown that production of steroids in the absence of ACTH and that in response to cholera toxin involve some effect of membrane cholesterol not dependent on cAMP. PMID- 3026783 TI - A phorbol ester, phorbol 12-myristate 13-acetate, and a calcium ionophore, A23187, can mimic the luteolytic effect of prostaglandin F2 alpha in isolated rat luteal cells. AB - To explore the possible role of protein kinase C and calcium in the luteolytic process, we treated luteal cells with a protein kinase C activator, the phorbol ester, phorbol 12-myristate 13-acetate (PMA), and with the calcium ionophore, A23187. Lower concentrations of PMA could clearly mimic the inhibitory, luteolytic effects of prostaglandin F2 alpha (PGF2 alpha) on LH-induced cAMP and progesterone production. A nontumor promoting phorbol ester, 4 alpha-phorbol 12,13-didecanoate, had no inhibitory effect, indicating a specific PMA effect. The calcium ionophore, A23187, also gave a marked inhibition of LH-induced cAMP and progesterone production. Hormone-stimulated adenylate cyclase activity was markedly impaired after preincubation of the cells with PGF2 alpha, PMA, or A23187, but no effect was seen when the substances were added to isolated membranes. In addition, the stimulation of progesterone production in the luteal cells with the cAMP analogs, 8-bromo-cAMP and (Bu)2cAMP, was almost totally abolished when PMA or A23187 was present. We conclude that PMA and A23187 in many ways mimic the effect of PGF2 alpha in luteal cells. The inhibition of steroidogenesis is partly dependent on depressed activity of the hormone sensitive adenylate cyclase, but also obtained by inhibiting steps distal to cAMP formation. Both points of action seem to be calcium and/or protein kinase C dependent. In contrast, higher concentrations of PMA markedly stimulated steroidogenesis without affecting the cAMP level, a stimulation not seen after incubation with 4 alpha-phorbol 12,13-didecanoate, suggesting again a specific PMA effect. The stimulation of steroidogenesis by higher concentrations of PMA seems to be specific, but the interpretation of this finding is unclear at present. In conclusion, PMA and A23187 mimic some of the luteolytic properties of PGF2 alpha, not only inhibiting the luteal cAMP system, but also by inducing lesions in the steroidogenic steps beyond the cAMP system. PMID- 3026784 TI - Cyclic hormonogenesis in gray collie dogs: interactions of hematopoietic and endocrine systems. AB - Cortisol, ACTH, T4, and T3 concentrations were determined in six cyclic hematopoietic (CH) dogs to determine if hormonal cycles were a feature of this hematopoietic disorder. It was determined that there were 12- to 14-day cycles of these hormones in CH dogs. Plasma cortisol and ACTH concentrations peaked 4-8 days after the onset of neutropenia and were concurrent with peak neutrophil counts. The peak ACTH concentration occurred 1-2 days before or concurrent with the cortisol peak concentration. T4 and T3 peak concentrations were opposite the cortisol cycles, and maximal concentrations were seen when cortisol levels were lowest. ACTH, GH-releasing factor and TRH response tests were performed in the CH dogs. No frank deficiencies in hormone production were seen with the ACTH, GH releasing factor, and TRH responses in CH dogs relative to those in normal dogs. It was concluded that cyclic hormonogenesis is a central feature of CH disease. These findings are the first demonstration of extrahematopoietic system cyclicity in this rare disease and suggest the presence of common regulatory factors in the hematopoietic and endocrine systems. PMID- 3026782 TI - In vivo association of [125I]-insulin with a cytosolic insulin-degrading enzyme: detection by covalent cross-linking and immunoprecipitation with a monoclonal antibody. AB - A cytosolic insulin-degrading enzyme (Mr = 110,000) was found to be cross-linked to [125I]-insulin in intact human hepatoma cells, HepG2, incubated with the hormone and treated with the bifunctional cross-linker, disuccinimidyl suberate. The labeling of this protein was greatly increased by concurrent treatment of the cells with N-ethylmaleimide, to the extent that the amount of [125I]-insulin cross-linked to the enzyme in these cells was approximately 20 to 50% that cross linked to the insulin receptor. The labeling of the insulin-degrading enzyme required the prior interaction of [125I]-insulin with its receptor as well as a temperature- and energy-dependent processing of the hormone. The present work therefore supports a role for this protease in the cellular processing of insulin. PMID- 3026786 TI - Phorbol myristate acetate inhibits alpha 1-adrenergically but not thyrotropin regulated functions in FRTL-5 rat thyroid cells. AB - The iodination of thyroglobulin and the formation of thyroid hormones are regulated by alpha 1-adrenergic agents as well as TSH in rat FRTL-5 cells. The regulatory effects of the alpha-1-adrenergic agents and TSH on both of these processes are associated with an increase in cytosolic Ca2+ and an increase in that component of iodide efflux that is representative of the movement of iodide from the thyroid cell into the follicular lumen. When FRTL-5 cells are preincubated with phorbol myristate acetate (PMA) for at least 3 min, the norepinephrine-stimulated changes in cytosolic Ca2+ levels and iodide efflux are inhibited. In contrast, PMA pretreatment has no effect on iodide efflux and actually enhances the changes in cytosolic Ca2+ induced by TSH. Phorbol myristate acetate pretreatment has no effect on TSH-stimulated cAMP-mediated iodide uptake in FRTL-5 cells, nor does it affect the binding parameters of the alpha 1 adrenergic receptor antagonist prazosin. These data suggest that protein kinase C is involved in a feedback mechanism regulating alpha 1-adrenergic but not TSH induced changes associated with the iodination of thyroglobulin and the formation of thyroid hormones; and that this feedback effect occurs after the step of ligand binding but before the increase in cytosolic Ca2+ induced by the alpha 1 adrenergic agents. PMID- 3026785 TI - Synergism between systemic corticotropin-releasing factor and arginine vasopressin on adrenocorticotropin release in vivo varies as a function of gestational age in the ovine fetus. AB - In sheep, parturition is associated with maturation of fetal pituitary-adrenal function, and with rises in the concentrations of ACTH and cortisol (F) in fetal plasma. We examined the hypothesis that pituitary ACTH output in response to arginine vasopressin (AVP) and CRF separately and together might change during late pregnancy as a function of fetal age. Fetal sheep were chronically catheterized, and bolus iv injections of equimolar AVP, CRF, AVP plus CRF, or saline (controls) were given on days 110-115, 125-130, and 135-140 of gestation. AVP evoked significant rises in plasma ACTH on days 110-115 and 125-130, but not on days 135-140. After AVP, the peak plasma concentrations of ACTH were attained at 5-10 min, and basal (preinjection) values were reestablished by 30-60 min. After CRF treatment, plasma ACTH rose progressively throughout the 240 min of the study. Evidence was obtained in support of an increase in pituitary responsiveness to CRF between days 110-115 and 125-130 and a decrease in response on days 135-140, when basal F concentrations were higher. The ACTH response to AVP, relative to that to CRF, was greatest in the youngest fetuses. On days 110 115 only, CRF and AVP showed a synergistic response in ACTH output, especially during the first 30 min after agonist injection. Plasma F rose in response to the changes in endogenously released ACTH in a manner consistent with progressive fetal adrenal maturation between days 110-140 of pregnancy. We conclude that in vivo the ovine fetal pituitary responds separately and synergistically to AVP and CRF on days 110-115 of gestation, but the relative role of AVP in stimulating ACTH release decreases with progressive gestational age. PMID- 3026787 TI - Measurement of 1,25-dihydroxyvitamin D3 receptor turnover by dense amino acid labeling: changes during receptor up-regulation by vitamin D metabolites. AB - We have previously demonstrated that 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] receptors are up-regulated by exposure to 24,25-(OH)2D3 and other vitamin D metabolites in several mammalian cell cultures systems as well as in vivo in rats. The goal of the current study was to determine how changes in receptor turnover could give rise to increased receptor levels after treatment with 24,25 (OH)2D3, whether by an increased rate of synthesis or a decreased rate of degradation. Receptor turnover was studied by the dense amino acid shift technique in cultured pig kidney cells (LLC-PK1). In control cells, the half-life of the 1,25-(OH)2D3 receptor was estimated to be 4.3 +/- 0.4 h, with a degradation rate constant (kD) of 0.16 +/- 0.01 h-1. In parallel experiments, cells were treated with 24,25-(OH)2D3 for about 48 h to reach a new up-regulated steady state before the measurement of receptor turnover. In up-regulated cells, the receptor half-life was prolonged to 8.9 +/- 1.0 h and the kD was 0.07 +/- 0.01 h-1. Assuming a steady state, the values obtained for total receptor levels and the degradation rate allow calculation of the synthesis rate (kS) at equilibrium. Control cells synthesized receptors at the rate of 3.4 +/- 0.4 vs. up-regulated cells at 4.9 +/- 0.5 fmol mg protein-1 h-1. In conclusion, the data indicate that up-regulation of receptors after exposure of cells to vitamin D metabolites results predominantly from a prolongation of receptor half-life in addition to a relatively smaller increase in the rate of receptor synthesis. PMID- 3026789 TI - The adenosine triphosphate-dependent calcium pump in rat small intestine: effects of vitamin D deficiency and cell isolation methods. AB - Duodenal villus cells from vitamin D-deficient (-D) and replete rats (+D) were isolated either in citrate buffers according to the method of Stern or by vibration in EDTA-containing solutions according to the method of Harrison and Webster. Basolateral plasma membrane vesicles (BLMV) were purified, and active Ca2+ transport was studied. The rates of ATP-dependent Ca2+-transport in BLMV from -D and +D animals were 2.6 +/- 1.0 and 10.6 +/- 0.6 nmol Ca2+/min X mg protein when cells had been isolated in citrate buffers. These transport rates were 9.2 +/- 0.7 and 9.6 +/- 0.5, respectively, when cells were isolated by vibration. The specific activities of other enzyme markers and vesicle parameters, such as the degree of resealing or purification factors, were not influenced by the cell isolation procedure. Addition of lipase and protease inhibitors to the citrate buffer or fasting the animals before death increase the active Ca2+ transport rates in BLM from -D rats to control levels. These results indicate that the ATP-driven Ca2+ pump in duodenal plasma membranes from vitamin D-deficient rats is more prone to inactivation during enterocyte isolation procedures. With the vibration technique, six populations of cells could be obtained that are sequentially released from villus tip to crypt base. A similar villus-crypt gradient of active Ca2+ transport was present in duodenal BLMV from D and +D animals. In ileal BLMV from vitamin D-deficient rats a significantly lower Ca2+ transport rate was found compared to that in +D animals, even when villus cells were isolated by vibration. It is thought that the mechanisms by which 1,25-dihydroxyvitamin D3 regulate transcellular Ca2+ fluxes in duodenum and ileum must be different. PMID- 3026788 TI - Rabbit myometrial oxytocin and alpha 2-adrenergic receptors are increased by estrogen but are differentially regulated by progesterone. AB - Both myometrial oxytocin and alpha 2-adrenergic receptors are induced by estrogen. To compare the regulation of these two receptor populations by progesterone, we measured myometrial receptor concentration in ovariectomized steroid-treated and in pregnant rabbits. To control for the effects of estrogen withdrawal, we used concomitant rather than sequential presentation of estrogen and progesterone in ovariectomized rabbits. Estradiol increased both myometrial oxytocin and alpha 2-adrenergic receptor concentrations in ovariectomized rabbits after 8 days of treatment. Simultaneous progesterone administration during the last 4 days of estradiol treatment reversed the induction of oxytocin, but not alpha 2-adrenergic, receptors. Similarly, administration of the antiprogestin RU 38486 to pregnant rabbits on day 27 of gestation resulted in premature delivery and evoked an increase in myometrial oxytocin receptor concentration mimicking that observed at term (day 31). However, RU 38486 did not significantly affect alpha 2-adrenergic receptor concentration. Our data provide further support for involvement of oxytocin receptors in parturition, but do not indicate a comparable function for myometrial alpha 2-adrenergic receptors. PMID- 3026790 TI - 1,25-Dihydroxyvitamin D3 induces the synthesis of vitamin D-dependent calcium binding protein in cultured chick kidney cells. AB - A 28,000 mol wt vitamin D-dependent calcium-binding protein (CaBP), first isolated from avian intestine, has recently been shown to exist in kidney and other tissues. To study the mechanism regulating the production of renal CaBP, we used primary cultures of chick kidney cells as an in vitro model. Renal cortical tubules isolated from vitamin D-deficient chicks were grown in serum-free, hormone-supplemented medium. Confluent cells were epithelioid and expressed CaBP, as demonstrated by immunocytochemistry and a specific RIA. 1,25-Dihydroxyvitamin D3 [1,25-(OH)2D3] increased cellular CaBP content in a dose-dependent manner (10( 10)-10(-7) M) from basal concentrations of 50-240 ng/mg protein to maximal concentrations of 600-1200 ng/mg protein 48 h after dosing. Cycloheximide (2 microM) inhibited 1,25-(OH)2D3 induction of CaBP, indicating that the mechanism requires new protein synthesis. Western blotting of cell extracts confirmed the identity of the inducible protein as the 28,000 mol wt CaBP. 25-Hydroxyvitamin D3 and 24,25-dihydroxyvitamin D3 were effective but somewhat less potent inducers of CaBP, whereas vitamin D3 was without significant effect. The activities of 25 hydroxyvitamin D3 and 24,25-dihydroxyvitamin D3 are attributed to their rapid conversion by kidney cells to 1 alpha-hydroxylated metabolites. The dose-response profiles for two additional bioresponses in these cells, namely induction of 24 hydroxylase and inhibition of 1 alpha-hydroxylase activity, were comparable to those for CaBP induction. To our knowledge, this is the first description of a cell culture system that exhibits 1,25-(OH)2D3-inducible CaBP in vitro. This model should permit the study of certain aspects of the physiology of 1,25 (OH)2D3 and CaBP in kidney that are not possible in vivo. PMID- 3026791 TI - Changes in the ratio of bioactive to immunoreactive adrenocorticotropin-like activity released by pituitary cells from ovine fetuses and lambs. AB - We have compared the biological (B) and immunological (I) ACTH-like activities (ALA) released by short term cultured pituitary cells from fetal (63-144 days old) and young lambs (7-150 days old) both under basal conditions, and when stimulated by ovine(o)CRF-(1-41) or arginine vasopressin (AVP), separately or in combination. The bioassay was based on the production of corticosteroids by cultured adrenal cells from adult sheep. The standard used in both the RIA and the bioassay was synthetic ACTH-(1-24). Under every condition, curves for dilution of the incubation media were parallel to the standard curves in both the RIA and the bioassay. When expressed per 5 X 10(5) pituitary cells, B-ALA output increased with gestational age and remained high after birth. During fetal life, B-ALA released was always lower than I-ALA. However, the B/I ratios increased with gestational age from 0.29 +/- (SE) 0.04 at 63 days to 0.70 +/- 0.04 at 144 days. Conversely, for postpartum animals, the B/I ratios were not different from 1, except in the case of stimulation by AVP alone, when they were significantly higher than 1.A 4-day treatment of pituitary cells from 120- to 126-day-old fetuses by AVP or cortisol resulted in an 1.5-fold enhancement of the B/I ratio of the ALA secreted under AVP stimulation. These results indicate that during development of the sheep I-ACTH levels are probably not representative of the corticotropic activity released by the pituitary. Furthermore, they suggest that the prepartum increase in fetal cortisol does result, in part, from an enhanced tropic drive to the fetal adrenal. PMID- 3026792 TI - Insulin generates an enzyme modulator from hepatic plasma membranes: regulation of adenosine 3',5'-monophosphate phosphodiesterase, pyruvate dehydrogenase, and adenylate cyclase. AB - Some of the acute actions of insulin may be mediated by the intracellular generation of a chemical substance that modulates certain enzymes. Such a substance has been identified which is released from liver plasma membranes after exposure to insulin. This substance was purified on sequential ion exchange, reverse phase, and gel permeations columns. The purified substance modulated the activities of cAMP phosphodiesterase, adenylate cyclase, and pyruvate dehydrogenase. The activities that modulated each of these enzymes exhibited singular chromatographic behavior and sensitivity to a variety of chemical reagents. Each activity was also produced by treatment of membranes with a phosphatidylinositol-specific phospholipase C. These results suggested that each of the enzyme-modulating activities was due to a single complex carbohydrate substance which contained inositol, phosphate, glucosamine, and other monosaccharides. The actions of this substance on these three enzymes mimicked those of insulin, suggesting that the release of this enzyme modulator might play a role in mediating some of the actions of the hormone. PMID- 3026793 TI - Selective photolabeling of high and low affinity binding sites for vasoactive intestinal peptide (VIP): evidence for two classes of covalent VIP-receptor complexes in intestinal cell membranes. AB - The biological activities of VIP derivatives as photoaffinity labels are described. The derivatives were obtained by VIP modification with azido phenyl glyoxal at arginyl residues 12 and 14 ([Az Bz Arg12-14]VIP) or only at position 14 ([Az Bz Arg14]VIP). The first derivative exhibited pronounced hydrophobic properties. The level of cAMP produced in HT 29 cells by this derivative represented 15% of that obtained by VIP at equimolar concentration (10(-10) M). The second derivative ([Az Bz Arg14]VIP) was synthesized by a new procedure, in which amino acids of VIP buried in the active site of the receptor were protected from azido phenyl glyoxal. This analog retained a high binding affinity for receptor (Kd, 0.5 nM for [125I-Tyr,Az Bz Arg14]VIP vs. 0.1 nM for [125I] VIP) and was found to be biologically active, as judged by stimulation of adenylate cyclase activity (production of cAMP was 120 pmol/10(6) cells vs. 140 pmol for VIP at 10(-10) M). Photolysis of this analog in the presence of HT 29 cell membranes resulted in a stimulation of adenylate cyclase which persisted in spite of repeated washings. Our findings indicate that [125I-Tyr,Az Bz Arg14] VIP binds covalently to active sites to form an active peptide-receptor complex. When low affinity binding sites were specifically photolabeled using a selective protection of high affinity sites by incubating membranes with 10(-10) M VIP for 5 min, the derivative did not stimulate adenylate cyclase activity. This suggests that VIP acts through a high affinity site to produce the biological activity and that the functional relevance of low affinity sites is unclear. PMID- 3026794 TI - Identification and characterization of two classes of receptors for oxytocin and vasopressin in porcine tunica albuginea, epididymis, and vas deferens. AB - The neurohypophysial hormones oxytocin (OT) and vasopressin (VP) are involved in the regulation of the contractility of the male genital tract in several animal species. We investigated the presence of specific binding sites for [3H]OT and [3H]arginine VP (AVP) in membranes prepared from tunica albuginea, epididymis, and vas deferens from prepubertal pigs 2-16 weeks of age. Membranes were incubated with [3H]OT and [3H]AVP in the presence or absence of the corresponding unlabeled peptides. Binding equilibrium was reached in 60 min at 22 C. Millimolar concentrations of Mg2+ increased the specific binding of both ligands. Analysis of families of self- and cross-displacement curves using the computer program LIGAND clearly demonstrated that two classes of binding sites were present in all tissues investigated. The first class of sites, designated the OT site, shows high affinity for OT, AVP, lysine vasopressin, arginine vasotocin, the selective OT agonists [Thr4,Gly7]OT and [Asu1,6]OT, and the OT antagonists derived from ornithine vasotocin (OVT), namely d(CH2)5Tyr(Et)OVT and dEt2OVT. The second class of sites, designated the VP site, shows high affinity for AVP, lysine vasopressin, arginine vasotocin, and the selective V1 antagonist d(CH2)5Tyr(Me)AVP. The V2 agonist [1-deamino,4-valine]8-D-AVP shows low affinity for both sites. Isotocin, desglycinamide [Arg-8]AVP and tocinoic acid were ineffective in displacing [3H]AVP or [3H]OT. The highest density of OT receptors was found in tunica albuginea and epididymis, whereas the highest density of AVP receptors was found in vas deferens. Adenylate cyclase was not activated in any of the tissues studied by concentrations of AVP or OT up to 100-fold greater than their Kd values. This is the first demonstration and pharmacological characterization of specific OT and V1 VP receptors in the tunica albuginea, epididymis, and vas deferens. The recent demonstration of high local concentration of neurohypophysial hormones in the gonads of several mammals support a physiological role of these OT and VP receptors in regulation of the motility of the male genital tract. PMID- 3026796 TI - Dietary fibers and heavy metal retention in the rat. AB - The metal-binding capacities of some gel-forming polysaccharides and other substances have been investigated in vitro in an attempt to relate their metal binding properties to the retention of dietary Pb and Cd in vivo. In equilibrium dialysis systems, aqueous solutions of alginic acid, pectin, agar, and carrageenan (1 g fiber/100 ml) all bound Pb and Cd to varying degrees. Alginic acid had the greatest binding capacity for Pb (50 micrograms Pb bound/mg fiber) and carrageenan for Cd (9.3 micrograms Cd bound/mg fiber). Addition of any one of these fibers, or indulin or glucuronic acid to the diet increased the tissue retention of one or both of the metals. Only cellulose supplementation reduced the retention of both Pb and Cd. Carrageenan decreased that of Pb and increased that of Cd. In another experiment alginic acid was shown to increase Pb retention in rats even when present at fairly low dietary concentrations (1 g/kg). PMID- 3026795 TI - The role of calmodulin antagonists on steroidogenesis by fetal zone cells of the human fetal adrenal gland. AB - The adrenal gland of the human fetus (HFA) is relatively large compared to that of the adult and exhibits an extremely high rate of steroidogenesis both in vivo and in vitro. The fetal zone cells make up 80-85% of the volume of the HFA and are the major site of steroid production during fetal development. We have recently demonstrated that calcium is involved in the regulation of steroidogenesis in fetal zone cells of the HFA. There is considerable evidence that many actions of calcium within cells are mediated by the calcium-binding protein calmodulin. The purpose of the present investigation was to determine if calmodulin also plays a role in HFA steroidogenesis. To investigate this possibility, the fetal zone was dissected from fetal adrenals of first and second trimester human abortuses. After collagenase digestion of the tissue, dispersed fetal zone cells were maintained in a Krebs-Ringers medium at 37 C for a 3-h incubation. Cells were incubated with and without ACTH (10(-8) M) in the presence of the calmodulin inhibitors trifluoperazine (TFP), chlorpromazine (CPZ), and calmidazolium (CAL) at concentrations of 5-100 microM. The media were assayed for contents of dehydroepiandrosterone sulfate (DS), cortisol (F), pregnenolone, and cAMP by RIA. The addition of ACTH stimulated F secretion 5- to 10-fold compared to that in control fetal zone cells. DS secretion increased up to 5-fold and pregnenolone about 2-fold in the presence of ACTH compared to values in control cells. ACTH also stimulated cAMP secretion by 10-fold compared to that in control cells. The addition of TFP, CPZ, and CAL significantly inhibited ACTH-stimulated DS, F, and pregnenolone secretion in a dose-related fashion to near-control levels. We observed that TFP, CPZ, and CAL inhibited cAMP accumulation as well as Bu2cAMP-stimulated steroid secretion. The metabolism of 22R-hydroxycholesterol to pregnenolone was inhibited by TFP and CPZ, but not by CAL. These studies suggest that calmodulin plays a role in regulating steroidogenesis in fetal zone cells of the HFA. PMID- 3026797 TI - Clearance and dimensional changes of crocidolite asbestos fibers isolated from lungs of rats following short-term exposure. AB - Previous studies in this laboratory have demonstrated fiber clearance and dimensional changes in chrysotile asbestos using a rat inhalational model of short-term exposure. The purpose of the present study was to determine whether or not similar changes occurred in crocidolite asbestos fibers isolated from the lungs of rats at various intervals after termination of exposure. Fibers were recovered on a membrane filter using a sodium hypochlorite digestion concentration technique, and the numbers and dimensions of the fibers assessed using scanning electron microscopy. The mass of crocidolite asbestos retained in the lung was then calculated. Of the respirable fraction, 19% was deposited in the lungs, and 25% of this amount was still present 1 month after exposure. These values are similar to the 23% deposition and 19% retention rates for chrysotile determined in our previous study. There was a progressive increase in mean fiber length with time postexposure (P less than 0.05), but no significant changes in the diameter of the population of crocidolite fibers retained in the lung. Thus it appears that the tendency for longer fibers to be retained within lung tissue is a characteristic shared by serpentine and amphibole asbestos fibers, whereas longitudinal splitting with progressive decrease in mean fiber diameter in vivo occurs primarily with the serpentine fibers. PMID- 3026798 TI - Erythrocyte membrane ATPases in diabetes: effect of dikanut (Irvingia gabonensis). AB - The levels of the three ATPases found in the erythrocyte membrane of diabetic patients were significantly lower than normal subjects. The distribution of the enzymes was also different. Na+,K+-ATPase and Mg2+-ATPase reflected the status of blood glucose more than Ca2+-ATPase. The ratio between two of the ATPases was sensitive to glycemic response. When dikanut, a viscous preparation, was fed to diabetics for 4 weeks, blood glucose became normal and the activities of the three ATPases increased significantly. The ratio among the enzymes also approached that of normal subjects. A relationship was found between the blood glucose level and erythrocyte membrane ATPases which, if linked to insulin binding or level, may provide a rapid inexpensive assay in diabetes research. PMID- 3026799 TI - Eicosanoids and equine leucocyte locomotion in vitro. PMID- 3026800 TI - Detection of adenovirus-specific immunoglobulin A in tears from patients with keratoconjunctivitis. PMID- 3026801 TI - Characterization of an isogenic set of methicillin-resistant and susceptible mutants of Staphylococcus aureus. AB - A low affinity penicillin binding protein (PBP2') present in methicillin resistant strains but absent in isogenic sensitive strains of Staphylococcus aureus is responsible for the phenotypic expression of the methicillin resistance. An additional factor controlling methicillin resistance is known to map on the chromosome at the insertion site omega 2003 (chr::Tn551). The influence of this factor on PBP2' synthesis was analysed. Isogenic methicillin sensitive and resistant mutants derived from a Staphylococcus aureus strain carrying methicillin resistance factors and the omega 2003 (chr::Tn551) insertion were characterized by population analysis and restriction analysis of the DNA surrounding the Tn551 insertion site, and their PBP2' content was determined. Both methicillin resistant and sensitive mutants produced levels of PBP2' similar to those of the original methicillin resistant parent. The mere presence of this PBP2' could therefore not be solely responsible for the phenotypic expression of methicillin resistance. The nature and site of action of the additional factor determined by the omega 2003 (chr::Tn551) site controlling expression of methicillin resistance still remains unknown. PMID- 3026802 TI - Conversion of a homogeneously methicillin-resistant strain of Staphylococcus aureus to heterogeneous resistance by Tn551-mediated insertional inactivation. AB - Plasmid pRN3208, thermosensitive for replication, and carrying the erythromycin transposon Tn551, was used for insertional inactivation of methicillin resistance in a highly and homogeneously resistant strain of Staphylococcus aureus. Two kinds of insertionally inactivated cells were obtained. Cultures of the major class contained highly methicillin resistant cells with a frequency of about 10( 3) to 10(-4), produced DNA with methicillin resistance transforming activity, and also produced penicillin binding protein 2a, the 78 kd low affinity penicillin binding protein characteristic of methicillin resistant Staphylococcus aureus, in apparently normal quantities. The single member of class B had no detectable methicillin resistant cells (less than 10(-8)) with an MIC greater than 1 micrograms/ml, contained no DNA with methicillin resistant transforming activity and no penicillin binding protein 2a. The data suggest that in the class A cells insertional inactivation did not affect the structural gene(s) of methicillin resistance but a regulatory locus or loci needed for the homogeneous expression of resistance. PMID- 3026803 TI - Molecular typing of methicillin and gentamicin resistant Staphylococcus aureus in Dublin. AB - The high incidence of infection in Dublin hospitals caused by non-typable strains of methicillin- and gentamicin-resistant Staphylococcus aureus (MGRSA) has created an important epidemiological problem as conventional methods of sub dividing these organisms have not been useful. This report describes a novel approach to the typing and analysis of MGRSA strains, particularly non-typable isolates, by comparing restriction endonuclease HindIII digest patterns of total cellular DNA; and by using Southern hybridization analysis to detect size variations or polymorphisms in restriction endonuclease cleavage fragments within small regions of the chromosome. Non-typable MGRSA strains and isolates belonging to two phenotypically related groups of phage-type 77 and 77/84 strains were readily subdivided on the basis of molecular size differences in high molecular weight DNA fragments generated by the enzyme HindIII. Restriction endonuclease fragment size polymorphisms were readily detected in many of the non-typable strains tested in hybridization experiments, and these were used for strain sub division. Both techniques were useful tools for the separation of closely related MGRSA strains. PMID- 3026804 TI - Kinetics of the modulation of chloroplastic fructose-1,6-bisphosphatase activity by thioredoxin fb. AB - The inactivation of reduced chloroplast fructose-bisphosphatase by oxidized thioredoxin fb has been studied during the enzyme reaction along the principle of Tian and Tsou [Biochemistry (1982) 21, 1028-1032]. A minimum model for this process is presented and its kinetic and equilibrium parameters have been determined. Thioredoxin fb binding to the enzyme is fast relative to catalysis and product desorption. Under quasi-equilibrium conditions oxidized thioredoxin is a non-competitive inhibitor of the enzyme reaction and must bind to a regulatory 'thioredoxin site'. The slow deactivation is thermodynamically favoured, and as expected from binding data, slowed down by the presence of substrate, fructose bisphosphate. The desorption of thioredoxin fb from the enzyme is extremely slow and this small protein may be regarded as a 'regulatory' subunit of fructose-bisphosphatase. PMID- 3026805 TI - Conformational change accompanies redox reactions of the tetraheme cytochrome c 554 of Nitrosomonas europaea. AB - Cytochrome c-554 of the ammonia-oxidizing chemolithoautotropic bacteria is thought to mediate electron transfer from hydroxylamine oxidoreductase to a terminal oxidase and/or to ammonia monooxygenase. The cytochrome has four c hemes which interact magnetically and have the same redox potential. We report that the kinetics of reduction of ferric cytochrome c-554 by dithionite or the oxidation of ferrous cytochrome c-554 by O2 or H2O2 are complex and multiphasic. Transient rapid-scan difference spectra indicate discrete maxima at approximately 418 nm, 425 nm and 432 nm. Absorbance changes at all three difference maxima appear to occur in all kinetic phases, although not in equal amounts for each wavelength. Reduction by 20 mM dithionite was biphasic. At pH 7.5 the first phase, which involved approximately 50% of the total absorbance change, had a rate constant (20 degrees C) of 140 s-1 and energy of activation of 20 kJ X mol-1. The slow phase had a rate constant 0.43 s-1 and a relatively high energy of activation, 87 kJ X mol-1, suggesting that a change in protein configuration accompanied the reaction. As the pH of the solution increased, the rate constant for both phases decreased and the fraction of absorbance change in the rapid phase increased. Oxidation of ferrous cytochrome c-554 by O2 involved a discrete rapid phase with a rate constant of 14 s-1, accounting for 6% of the absorbance. The remainder of the reaction was multiphasic with rate constants in the range 0.1-0.01 s-1. With H2O2 as the oxidant, the rapid phase involved 39% of the change in absorbance with a rate constant of 19 s-1. The remainder of the reoxidation was multiphasic with rate constants ranging over 0.4-0.01 s-1. PMID- 3026806 TI - Characterization of a new endoglucanase from Erwinia chrysanthemi. AB - The structural gene coding for a new endo-beta-1,4-glucanase of Erwinia chrysanthemi strain 3665, previously identified in a cosmid library, was subcloned into pUC18. The gene is expressed from a 1.9 X 10(3)-base-pair insert and its direction of transcription was determined. The properties of the gene product purified from cell-free extracts of Escherichia coli have been studied. The purified protein has an endoglucanase activity but is significantly different from the major endoglucanase Z secreted by E. chrysanthemi strain 3665. The new enzyme was designated as endoglucanase Y and the related gene celY. In E. coli, most of the endoglucanase activity was found in the periplasmic space. PMID- 3026807 TI - Enhancer-mediated activation of a growth-regulated promoter. AB - We have demonstrated that the collagen alpha 2 type 1 promoter inserted in an expression vector, behaves as a growth-regulated promoter, which is consistent with previous observations that collagen synthesis is growth-regulated in vivo. In contrast, the activity of the H2-K or the simian virus 40 early promoters does not seem to be affected by the rate of cell proliferation. The insertion of a polyoma enhancer 5' or 3' to the collagen transcription unit activates the collagen alpha 2 type 1 promoter, by a threefold greater factor in slowly growing cells compared to cells growing exponentially. These results show that enhancers can also function in slowly proliferating cells and activate the normally low activity of a promoter in these cells. PMID- 3026808 TI - Lateral proton conduction at a lipid/water interface. Effect of lipid nature and ionic content of the aqueous phase. AB - Fast lateral proton conduction was observed along the lipid/water interface using a fluorescence technique. This conduction can be detected for a large number of lipids, both phospholipids and glycolipids. The efficiency of the proton transfer is dependent on the molecular packing of the host lipid at a given surface pressure. The proton conduction which is present in the liquid expanded state is abolished by the transition to the liquid condensed state. The proton transfer is affected slightly by the ionic content of the aqueous subphase except in the case of calcium which can inhibit the conduction along phosphatidylglyceroethanolamine. We suggest that the transfer of the protons occurs along a bidimensional hydrogen-bond network formed from the polar head groups, their water molecules of hydration and the water molecules which are intercalated between the lipid molecules. PMID- 3026809 TI - Effects of pH and fructose 2,6-bisphosphate on oxidized and reduced spinach chloroplastic fructose-1,6-bisphosphatase. AB - This report describes the effects of pH and fructose 2,6-bisphosphate (an analog of fructose 1,6-bisphosphate) on the activity of oxidized and reduced fructose 1,6-bisphosphatase from spinach chloroplasts. Studies were carried out with either fructose 1,6-bisphosphate, the usual substrate, or sedoheptulose 1,7 bisphosphate, an alternative substrate. The reduction of the oxidized enzyme is achieved by a thiol/disulfide interchange. The pK values relative to each redox form for the same substrate (either fructose 1,6-bisphosphate or sedoheptulose 1,7-bisphosphate) are identical, suggesting the same site for both substrates on the active molecule. The finding that the analog (fructose 2,6-bisphosphate) behaves like a competitive inhibitor for both substrates also favours this hypothesis. The inhibitory effect of this sugar is more important when the enzyme is reduced than when it is oxidized. The shift in the optimum pH observed when [Mg2+] was raised is interpreted as a conformational change of oxidized enzyme demonstrated by a change in fluorescence. The reduced and oxidized forms have the same theoretical rates relative to both substrates, but the reduced form has an observed Vmax which is 60% of the theoretical Vmax while that of the oxidized form is only 37% of the theoretical Vmax. The reduced enzyme appears more efficient than the oxidized one in catalysis. PMID- 3026811 TI - Purification and characterization of a hygromycin B phosphotransferase from Streptomyces hygroscopicus. AB - A hygromycin B phosphotransferase activity from Streptomyces hygroscopicus has been highly purified by ammonium sulphate fractionation followed by affinity column chromatography through Sepharose-6B-hygromycin-B. The combined active fractions showed a single protein band (41 kDa) when subjected to polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate. When gel electrophoresis was performed under non-denaturing conditions, the single protein band promoted in situ phosphorylation of hygromycin B, indicating that this protein corresponded to the purified hygromycin B phosphotransferase. The enzyme has been purified 236-fold and approximate Km values of 0.56 microM for hygromycin B and ATP, respectively, were deduced. PMID- 3026810 TI - Cloning and expression in Escherichia coli of a hygromycin B phosphotransferase gene from Streptomyces hygroscopicus. AB - The Streptomyces hygroscopicus hyg gene encoding a hygromycin B phosphotransferase has been introduced into different sites of both the Escherichia coli plasmid pBR322 and the Escherichia coli-Saccharomyces cerevisiae shuttle vector YRp7. When this gene was inserted into the BamHI site of pBR322 and then cloned in E. coli phosphorylating activity was not detected, indicating that the hyg gene promoter was not functional in this bacterium. However, when the hyg gene was inserted into either the unique PstI site of pBR322 or into each of the two PstI sites of YRp7, phosphotransferase activity was observed. Analysis of the translation products from these constructions by coupled in vitro transcription-translation systems suggested that in all cases transcrition was regulated by a promoter not provided by the inserted hyg gene and that the synthesized polypeptide was identical to that present in S. hygroscopicus. PMID- 3026812 TI - Properties of a multimeric protein complex from chloroplasts possessing potential activities of NADPH-dependent glyceraldehyde-3-phosphate dehydrogenase and phosphoribulokinase. AB - A homogeneous multimeric protein isolated from the green alga, Scenedesmus obliquus, has both latent phosphoribulokinase activity and glyceraldehyde-3 phosphate dehydrogenase activity. The glyceraldehyde-3-phosphate dehydrogenase was active with both NADPH and NADH, but predominantly with NADH. Incubation with 20 mM dithiothreitol and 1 mM NADPH promoted the coactivation of phosphoribulokinase and NADPH-dependent glyceraldehyde-3-phosphate dehydrogenase, accompanied by a decrease in the glyceraldehyde-3-phosphate dehydrogenase activity linked to NADH. The multimeric enzyme had a Mr of 560,000 and was of apparent subunit composition 8G6R. R represents a subunit of Mr 42,000 conferring phosphoribulokinase activity and G a subunit of 39,000 responsible for the glyceraldehyde-3-phosphate dehydrogenase activity. On SDS-PAGE the Mr-42,000 subunit comigrates with the subunit of the active form of phosphoribulokinase whereas that of Mr-39,000 corresponds to that of NADPH-dependent glyceraldehyde-3 phosphate dehydrogenase. The multimeric enzyme had a S20,W of 14.2 S. Following activation with dithiothreitol and NADPH, sedimenting boundaries of 7.4 S and 4.4 S were formed due to the depolymerization of the multimeric protein to NADPH dependent glyceraldehyde-3-phosphate dehydrogenase (4G) and active phosphoribulokinase (2R). It has been possible to isolate these two enzymes from the activated preparation by DEAE-cellulose chromatography. Prolonged activation of the multimeric protein by dithiothreitol in the absence of nucleotide produced a single sedimenting boundary of 4.6 S, representing a mixture of the active form of phosphoribulokinase and an inactive dimeric form of glyceraldehyde-3-phosphate dehydrogenase. Algal thioredoxin, in the presence of 1 mM dithiothreitol and 1 mM NADPH, stimulated the depolymerization of the multimeric protein with resulting coactivation of phosphoribulokinase and NADPH-dependent glyceraldehyde-3 phosphate dehydrogenase. Light-induced depolymerization of the multimeric protein, mediated by reduced thioredoxin, is postulated as the mechanism of light activation in vivo. Consistent with such a postulate is the presence of high concentrations of the active forms of phosphoribulokinase and NADPH-dependent glyceraldehyde-3-phosphate dehydrogenase in extracts from photoheterotrophically grown algae. By contrast, in extracts from the dark-grown algae the multimeric enzyme predominates. PMID- 3026813 TI - Reconstitution of purified chicken gizzard 5'-nucleotidase in phospholipid vesicles. Evidence for its transmembraneous character and the existence of functional domains on both sides of the phospholipid bilayer. AB - 5'-Nucleotidase, purified to homogeneity from chicken gizzard using published procedures [Dieckhoff, J., Knebel, H., Heidemann, M. and Mannherz, H. G. (1985) Eur. J. Biochem. 151, 377-383] was incorporated into artificial phospholipid vesicles after prolonged dialysis against detergent-free buffer or by a gel filtration procedure. After dialysis the obtained liposomes exhibit a mean diameter of 80 nm and contain 5'-nucleotidase at random orientation, demonstrated by finding up to 50% of the total liposome-incorporated AMPase activity to be cryptic, i.e. could only be measured after their permeabilization by addition of detergent. By affinity chromatography a phospholipid vesicle fraction could be obtained containing almost exclusively cryptic AMPase activity, thus representing the inside-out orientation of 5'-nucleotidase. Comparative analysis of physiochemical and enzymatic properties of 5'-nucleotidase reveals differences between the detergent-solubilized and the liposome-incorporated 5'-nucleotidase including a changed accessibility of the enzyme to polyclonal and monoclonal antibodies. Binding and AMPase inhibition studies with different polyclonal antibodies strongly indicate to the existence of a cytoplasmic domain of chicken gizzard 5'-nucleotidase. F-actin appears preferentially to interact with the cytoplasmic domain of liposome-incorporated 5'-nucleotidase. PMID- 3026814 TI - Generation of a transmembrane gradient of Na+ in Methanosarcina barkeri. AB - A transmembrane Na+ gradient was generated by Methanosarcina barkeri during methanogenesis. The intracellular Na+ concentration amounted to approximately one fifth of the extracellular one. A secondary Na+/H+ antiport system was shown to be responsible for Na+ extrusion. This system could be inhibited by amiloride. In the presence of amiloride the delta pH across the cytoplasmic membrane increased and a transmembrane Na+ gradient could neither be generated nor maintained. The possible role of Na+ in the oxidation of methanol to the level of formaldehyde is discussed. PMID- 3026815 TI - [A comparison of two antigen strains of each of mouse hepatitis virus and mouse adenovirus for detection of complement fixation antibody in mouse and rat sera]. AB - Detection rates of complement fixation antibodies in mice and rats were compared between two antigen strains of each of mouse hepatitis virus (MHV) and mouse adenovirus (MAV). Among 66 and 47 naturally infected MHV-positive sera of mice (18 facilities) and rats (16 facilities) respectively, 17 mouse and 21 rat sera reacted with both Nu-67 and MHV-2 strains, but 49 mouse and 25 rat sera were positive to Nu-67 strain alone. Only one rat serum reacted with MHV-2 strain alone. In comparison with K87 and FL strains of MAV, all 8 positive mouse sera (3 facilities) reacted with K87 strain alone whereas out of 53 positive rat sera (20 facilities), 43, 6 and 4 sera reacted with K87 strain alone, with FL strain alone and with both the two strains, respectively. PMID- 3026816 TI - Prevalence of antibodies to Sendai virus and rotavirus in laboratory rabbits. AB - One hundred and sixty rabbit sera from 10 breeding colonies and 13 laboratory colonies were tested for antibodies to Sendai virus and rotavirus by enzyme linked immunosorbent assay (ELISA). Antibodies were detected to Sendai virus in 53% and to rotavirus in 81%, indicating the prevalence of these viral infections in laboratory rabbit colonies. PMID- 3026817 TI - A survey of Streptococcus pneumoniae, Streptococcus zooepidemicus, Salmonella spp., Bordetella bronchiseptica and Sendai virus in guinea pig colonies in Japan. AB - S. pneumoniae, S. zooepidemicus, Salmonella spp., B. bronchiseptica and Sendai virus were examined in a total of 45 guinea pig colonies (17 institutional and 28 breeder's colonies) of Hartley strain, and found to be positive in 6, 3, 5, 20 and 14 colonies, respectively. Sendai virus was highly prevalent among guinea pigs, showing so high rates positive as 80 to 100% of animals obtained from 11 of the 14 contaminated colonies. B. bronchiseptica was also shown to be prevalent within contaminated colonies by indication of rates positive more than 40% of animals examined in 14 of the 20 colonies. Infection rates of other 3 pathogens, however, ranged from lower than 20% to higher than 80% according to colonies. All strains of Salmonella isolated in this survey were identified as S. typhimurium and those of S. pneumoniae as serotype 19F. PMID- 3026818 TI - A serological survey on 15 murine pathogens in mice and rats. AB - A serological survey for 15 murine pathogens was performed on 269 mouse sera collected from 21 conventional and 12 barrier colonies, and on 376 rat sera collected from 21 conventional and 23 barrier colonies. Animals having an antibody against at least one of the antigens were contained in 81.0% of conventional and 16.7% of barrier mouse colonies and also in 81.0% of conventional and 43.5% of barrier rat colonies. Main contaminants were mouse hepatitis virus and Sendai virus in mice, and Sendai virus and pneumonia virus of mice in rats. Results also indicated that antibodies to Toolan's H-1, minute virus of mice and PVM were positive in mice from a considerable number of colonies and those to Kilham rat virus, Mycoplasma pulmonis and Toolan's H-1 were sometimes detected in rats, suggesting prevalences of these pathogens in mice and rats in Japan. PMID- 3026820 TI - Lithium inhibits terminal differentiation of erythroleukemia cells. Evidence for a pre-commitment 'priming' event. AB - Lithium has been found to be a novel inhibitor of the terminal differentiation of Friend murine erythroleukemia cells. A general method for the quantitative analysis of differentiation inhibitors has been developed and used to compare the site of inhibition by lithium with that by vanadate. Lithium inhibits the commitment to differentiation (K 1/2 approximately 10 mM) induced by DMSO (dimethylsulfoxide) at non-toxic concentrations that have only a small effect on the rate of proliferation. Inhibition is reversible and probably requires entry of Li+ into the cell, since it is blocked by high KCl in the medium. LiCl is most effective when present during the first 10 h of DMSO treatment, before commitment is initiated. Computer-assisted analysis of the kinetics of commitment demonstrate that inhibition by lithium is best described by including a lithium sensitive 'priming' event, which occurs with high probability prior to commitment. PMID- 3026819 TI - Effect of cytochalasins on cytosolic-free calcium concentration and phosphoinositide metabolism in leukocytes. AB - Cytochalasins are routinely used to stimulate a variety of functions in eukaryotic cells even though their precise mode of action remains to be elucidated. In the present work we used the fluorescent Ca2+ indicator quin2 to study the effect of various cytochalasins, cytochalasins A, B, C, D, E (CA, CB, CC, CD, CE) and dihydrocytochalasin B (dhCB) on the intracellular Ca2+ concentration ([Ca2+]i) in various types of leukocytes, viz, neutrophils and lymphocytes. In human neutrophils, cytochalasins increase [Ca2+]i mainly by releasing Ca2+ from membrane-bound, intracellular stores. Thus, in order to readily appreciate the effect of cytochalasins on [Ca2+ )i, these cells must be loaded with low intracellular quin2 concentrations. On the other hand, in peripheral blood lymphocytes, splenocytes and thymocytes, the increase in [Ca2+]i is predominantly due to an increased Ca2+ influx from the extracellular medium. In addition, we found that in neutrophils these drugs prolong the increase in [Ca2+]i induced by chemotactic peptides, probably by increasing the cell permeability to Ca2+. Finally, in thymocytes, cytochalasins potentiate the production of inositol phosphates induced by the polyclonal mitogen concanavalin A (conA). PMID- 3026821 TI - Cytoskeletal association of pp60src. The transforming protein of the Rous sarcoma virus. AB - Immunoferritin labelling methods have been employed to examine the distribution of the Rous Sarcoma virus (RSV)-transforming protein pp60src in the detergent resistant cytoskeleton of transformed cells. pp60src was found to be localized on actin microfilaments present in adhesion plaques, at adherens junctions between cells and also in microfilament bundles. This localization is consistent with the hypothesis that some of the morphological effects of transformation result from the interaction in situ of pp60src with microfilament-bound target proteins. PMID- 3026822 TI - Transfer of 'immortalizing' oncogenes into rat fibroblasts induces both high rates of sister chromatid exchange and appearance of abnormal karyotypes. AB - Cell lines established after transfer into FR3T3 rat fibroblast cells of 'immortalizing' oncogenes (plt gene (large T protein) of polyoma virus, v-myc gene of MC29 virus, rearranged forms of c-myc) exhibited increased rates of sister chromatid exchange (SCE). This was observed neither in cells which expressed one of the oncogenes responsible for the terminal stages of tumorigenic transformation (polyoma virus pmt (middle T protein), mutated ras genes), nor in cell lines carrying oncogenes of both types. Abnormal chromosome numbers were observed in cell lines expressing plt or myc, but not after transformation by pmt or ras oncogenes. PMID- 3026824 TI - Random integration of SV40 in SV40-transformed, immortalized human fibroblasts. AB - We have studied the relationship between immortalization of SV40-transformed human embryonic fibroblasts and their SV40 integration sites. From several independently transformed cell pools, we have isolated clones which do not harbor unintegrated SV40 DNA. We have analysed whole-cell DNA from these clones, using the Southern blot method. Our results suggest that no specific integration sites in the cellular genome exist which are a prerequisite for the immortalization process. Although some integration sites were found to be predominant in pre crisis clones, they could not be detected in the post-crisis clones. This suggests that none of these predominating sites is selected for during the crisis period. PMID- 3026823 TI - Identification of a surface protein of the rabbit blood platelet with high affinity for collagen. AB - Platelet membrane components adhering with high affinity to collagen fibers were studied by means of an affinity column in which fibrillar type I collagen was physically immobilized. Intact rabbit platelets in 1 mM EGTA adhered to the column but did not aggregate. Adhesion was dependent on the collagen concentration and on the number of platelets applied. Passage through the column without adhesion did not affect the potential for subsequent platelet binding. Surface-labelled whole platelets were passaged through this column, lysed in Triton and in SDS and labelled components adhering to the collagen were analysed on SDS-polyacrylamide gels. It was found that Triton lysis removed most of the major surface glycoproteins but left the cytoskeleton on the column. Subsequent SDS elution removed the cytoskeletal proteins along with the remaining major surface glycoproteins. The label left on the column could not be eluted with 8 M urea or up to 4 M NaCl. Collagenase digestion of the column collagen released a single surface glycoprotein of Mr 80,000. Limited chymotryptic digestion of the labelled platelets prior to their application to the column did not affect their binding. A radiolabelled band of the same molecular weight (MW) became bound to the collagen following passage of the chymotrypsin-treated platelets. This band was trypsin-sensitive following SDS-polyacrylamide gel electrophoresis (SDS PAGE). These results, along with other published evidence, suggest that at least one platelet membrane component, expressed on the surface of the unstimulated platelet, binds with high affinity to fibrillar type I collagen and is probably involved in platelet collagen recognition. PMID- 3026825 TI - In vivo cGMP levels in frog photoreceptor cells as a function of light exposure. AB - By employing a combination of highly sensitive radioimmunoassays and histochemical techniques, an in vivo time course of cGMP levels has been determined in the outer segment, photoreceptor cell and outer plexiform layers of frog retina. Frogs (Rana pipiens) were dark-adapted overnight and either frozen rapidly (approximately 3 sec) in liquid nitrogen or exposed to periods of light varying between 0.1 sec and 2 hr before freezing. Frozen retinal sections were cut, freeze-dried, and samples of individual layers dissected out and analysed for cGMP. In the outer plexiform layer, there was a 42% drop in cGMP concentration after 2 sec of light (250 ft candles) followed by a 34% rise after 2 min; a steep concentration gradient formed around the layer after the 2 min exposure. In both the outer-segment layer and photoreceptor-cell layer (which includes outer segments, inner segments and outer nuclear layers), cGMP levels declined from a dark value of 56 mumol kg-1 (dry) to 9 mumol kg-1 (dry) as a result of increasing exposure to several types of light source: levels appear to be primarily a function of total ft candle min. Cyclic GMP concentrations at the longest exposures (2 min with a fiber optic light source or 2 hr with fluorescent room light) reached identical minimum levels. In the outer segments, a 15% decrease in cGMP was observed after 0.1 sec of light exposure. Although the freezing time is too long to be able to say whether the 15% decrease in cGMP at the 0.1 sec exposure is involved in transduction, the low identical levels reached gradually after longer exposures appear to indicate that a light-induced biochemical adjustment in cGMP metabolism occurs over a relatively long time period separate from the msec time course of the transduction process. PMID- 3026826 TI - Cyclic nucleotide levels of the photosensitive irises of Bufo marinus and Lophius. AB - The sphincter pupillae muscle of several species contracts autonomously after light directly strikes the iris. The response is initiated by isomerization of rhodopsin in the sarcolemma. Light-induced release of CA2+ from internal stores is one step in the transduction process. The levels of cyclic AMP and cyclic GMP were measured after irises were stimulated by light. Stimulation by light does not produce measurable or consistent changes in the levels of either cyclic AMP or cyclic GMP within the photosensitive irises of at least two species, Bufo marinus and Lophius. PMID- 3026827 TI - Nuclear cataract: oxidative damage to the lens. PMID- 3026828 TI - Changes in activity and amount of active elongation factor 1 alpha in aging and immortal human fibroblast cultures. AB - Stoichiometrically estimated amounts of active elongation factor, EF-1 alpha, remain constant in serially passaged Phase II cultures of human fibroblasts, MRC 5, but decrease by 45% towards the end (Phase III) of their lifespan. Catalytic activity of EF-1 alpha is also reduced by 35% in Phase III old cells. The SV40 transformed immortal cell line MRC-5V2 has 30% higher levels of active EF-1 alpha without significant increase in its catalytic activity. Low-serum-associated G1 arrest of normal and transformed cells reduces amounts of active EF-1 alpha by 35% and 20%, respectively. Catalytic activity, however, is reduced rapidly only in G1 arrested normal cells and not in transformed cells. Even though the cell cycle-related changes are reversible both in normal and transformed cells, the age-related decline in amounts of active EF-1 alpha and its activity are irreversible and, most probably, crucial. PMID- 3026830 TI - Effects of "partial axotomy" upon synaptic function. AB - A lesion limited to the dorsal columns, at the level of L3-L4, was carried out in chronic cats. This operation produced a "partial axotomy" type of lesion on the ascending branches of Ia hind limb afferents. Two to six months after this operation, intracellular studies on L7-S1 motoneurons were carried out. Similar studies were done in normal animals. The peak amplitude and the rate of rise (dV/dt) of heteronymous EPSP's were studied during control conditions (sampling at 1 Hz) and during the post-tetanic potentiation produced by a 500 Hz tetanus (for 3 s). The analysis of these synaptic potentials makes us conclude that: The amplitude of the enlarged EPSP's, observed during PTP, seems to be linearly dependent on their amplitude during control (i.e., pre-tetanus) conditions. Judging by their amplitude, there is no difference between potentiated EPSP's of operated and normal animals. There is also a linear relationship between the rate of rise of EPSP's and their peak amplitude. The slope of this relationship becomes steeper after "partial axotomy", i.e., for a given EPSP amplitude, the dV/dt of its rising part is steeper in operated cats. This steeper slope is also present in EPSP's studied during PTP. The sharper rate of rise of EPSP's, induced by the "partial axotomy" of Ia fibers, would be the mechanism behind the larger monosynaptic reflex previously observed in these operated cats. PMID- 3026829 TI - Facilitation and inhibition of synaptic transmission in the spinal cord: an electroneurographic study in humans. AB - The neurographic activity evoked either by stimulation of the tibial nerve at the popliteal fossa or by percussion of the Achilles tendon has been recorded at lumbar and thigh levels, in order to find out whether conduction time, temporal dispersion and central delay of the neural volleys underlying the monosynaptic reflex (H or T) may change as a function of stimulus intensity; under facilitatory or inhibitory experimental conditions; "spontaneously", i.e. during the steady state. The reflexly evoked ventral root discharge (VRD) decreases in latency with increasing stimulus intensity up to the maximum reflex response in the absence of changes in afferent (thigh to spine) or efferent (spine to thigh) conduction times. Reduction of the central delay was greater with mechanical than electrical stimulation, probably due to the combined effect of spatial and temporal summation under the former experimental condition. The latency of the VRD related to the maximal H response was not further modified by supramaximal stimulus strengths. The Jendrassik manoeuvre caused a significant decrease in latency of the VRD, the opposite effect being observed during calf muscle vibration. A significant relationship between amplitude and latency of single VRDs could be demonstrated during the "steady state". Our data point to the existence of a positive correlation between the size of the motoneuronal pool activated by an afferent volley and speed of transmission in the reflex pathway, both during the "steady state" and under either facilitatory or inhibitory experimental conditions, provided that the test stimulus strength does not exceed the maximum reflex response (H or T). No detectable signs of peripheral dispersion of the VRD could be demonstrated, irrespective of the stimulus employed: this suggests that the axon diameters of the motoneurones contributing to the monosynaptic reflex fall within a fairly narrow distribution. PMID- 3026831 TI - The interneuronal nature of GABAergic neurons in the lateral geniculate nucleus of the rhesus monkey: a combined HRP and GABA-immunocytochemical study. AB - Neurons immunoreactive to a GABA antibody, located in the retrogradely labeled segment of LGN of the rhesus monkey obtained after massive injections of HRP in a sector of the striate cortex, were examined for HRP label in plastic semithin (1 micron) sections of LGN. In a total of 691 GABA(+) neurons sampled from the 6 main geniculate layers none was seen to contain HRP grains. In contrast, the vast majority of GABA(-) neurons were densely labeled with HRP reaction product. It is concluded from these results that GABAergic cells in macaque LGN do not project to the striate cortex, as previous studies have shown for equivalent cells in cat LGN, and therefore reasons are given to consider these cells as interneurons in this nucleus. In addition, several GABA(-) neurons which were also unlabeled with HRP were observed isolated in the midst of populations of neurons homogeneously labeled with this enzyme. These cells were mostly located at or near the interlaminar zones and some of them were substantially larger than the neighboring geniculostriate relay cells in the parvocellular subdivision of LGN. For these characteristics, these unlabeled somata are considered to correspond to the geniculate neurons projecting to prestriate cortical visual areas in the macaque described in several studies and which, the present results suggest, do not branch collateral axons to the striate cortex. PMID- 3026833 TI - Chronic infusion of agents that increase cyclic AMP concentration enhances the regeneration of mammalian peripheral nerves in vivo. AB - Our previous investigation indicates that forskolin, a robust activator of adenylate cyclase, promotes sensory nerve regeneration in amphibians. The present study was designed to determine if forskolin had a similar effect in mammals. We also wished to test the hypothesis that cyclic AMP modulates nerve regeneration by comparing the effects of chronically infused forskolin with the effects of infused dibutyryl cyclic AMP, 8-bromo cyclic AMP, and the phosphodiesterase inhibitor, theophylline. Our results indicated that all agents promoted some aspect of regeneration. The two which presumably generated the largest increase in cyclic AMP concentration, forskolin and 8-bromo cyclic AMP, had the most profound effect on axonal elongation. All agents decreased the time to sprout initiation, but theophylline produced the largest decrease and its effect was mimicked by caffeine, a methylxanthine with limited ability to inhibit phosphodiesterase. This suggests that sprout formation may be triggered by an increase in intraaxonal free Ca2+, possibly modulated by cyclic AMP. The role of cyclic AMP in axonal elongation remains to be determined, but may be associated with stimulation of protein synthesis in the nerve cell body. PMID- 3026834 TI - Giardia lamblia: detection and characterization of calmodulin. AB - Calmodulin was detected in Giardia lamblia by radioimmunoassay and cyclic AMP phosphodiesterase activation. This protein was purified to apparent homogeneity by fast protein liquid chromatography with a yield of 260 ng of calmodulin/mg of protozoan protein. Purity was established by gel electrophoresis, gel filtration, and ion exchange chromatography. The parasite calmodulin has properties characteristic of calmodulin isolated from other eukaryotes, e.g., an apparent molecular weight of 16.7 kD; activation in calcium dependent manner of bovine heart cyclic nucleotide phosphodiesterase; and sensitivity to known calmodulin antagonists. PMID- 3026835 TI - Trypanosoma cruzi: subcellular distribution of glycolytic and some related enzymes of epimastigotes. AB - The first six glycolytic enzymes in epimastigote Trypanosoma cruzi were shown to behave similarly during differential centrifugation, when maximum relative specific activity was found in the small granule fraction, and by isopycnic centrifugation, when the bulk of each activity coequilibrated on sucrose gradients with a modal density of 1.23 g/ml. All six showed substantial detergent latency in whole cell homogenates. Electron microscopic examination of fractions from a sucrose gradient with modal density 1.23 g/ml showed the presence of single membrane bound vesicles of diameter 0.2-0.8 micron. It was concluded that these six enzymes were contained in a microbodylike organelle, termed the "glycosome." Phosphoglucose isomerase (EC 5.3.1.9) also possessed substantial soluble activity. No microbody marker enzyme described in other sources could be detected. Peroxidase (EC 1.11.1.7) had an insignificant glycosomal component. Enzymes of amino acid and fatty acid metabolism were not detected in microbody fractions. Marker enzymes for the flagellar pocket and plasma membrane were suggested. PMID- 3026832 TI - Calcium and epileptogenesis. PMID- 3026836 TI - Inhibition of phorbol ester stimulated superoxide production by 1-oleoyl-2-acetyl sn-glycerol (OAG); fact or artefact? AB - OAG-stimulated superoxide (O2) production by HL-60 granulocytes showed enantiomeric specificity but reached a maximum of only 5% of that produced by either phorbol myristate acetate (PMA) or phorbol dibutyrate (PDBu). At 10-100 microM, OAG displaced specifically-bound [3H]PDBu from intact HL-60 cells by only 25%, suggesting limited cell penetration. OAG (10-100 microM) also inhibited PDBu stimulated O2 production by 25%; this inhibition was enantiomerically specific. However, at a lower concentration (3 microM), both enantiomers of OAG fully blocked O2 production stimulated by PMA (0.5 microM). This inhibition is probably artefactual, due to the hydrophobic PMA physically associating with OAG in the extracellular fluid. PMID- 3026837 TI - cDNA cloning and expression in E. coli of a plasminogen activator inhibitor (PAI) related to a PAI produced by Hep G2 hepatoma cell. AB - The human hepatoma line Hep G2 produces an acid- and SDS-sensitive plasminogen activator inhibitor (PAI). This protein has been previously purified and used to raise polyclonal antibodies. This antiserum has been used to isolate cDNA clones from a human placental lambda gt11 cDNA library. The immunologically positive clones were screened for expression of recombinant proteins which inhibit urokinase activity and form an inhibitor-enzyme complex with 125I-urokinase. Two positives (lambda PAI 11.1 and lambda PAI 14.1) have been obtained. The cDNA insert of the longer isolate (lambda PAI 14.1) consists of 1962 base pairs encoding the entire mature Hep G2 PAI and a 3'-noncoding region of 801 base pairs. The clone apparently lacks portions of 5'- and 3'-untranslated sequences. The translated amino acid sequence matches the sequence obtained for the mature Hep G2 PAI and consists of 379 amino acids with a molecular mass of 42 770 Da. Interestingly, this PAI clone is quite different from the placental-type PAI-2 sequence as expected, but matches the sequence of the endothelial-type PAI (PAI 1) reported to be acid-insensitive and SDS-enhancible. PMID- 3026838 TI - Evidence for a GTP-binding protein coupling thrombin receptor to PIP2 phospholipase C in membranes of hamster fibroblasts. AB - Two different methods were used to study directly alpha-thrombin modulation of polyphosphoinositide breakdown in membranes prepared from Chinese hamster lung (CHL) fibroblasts. In the first one we labelled the lipid pool by incubating the intact cells with myo-[3H]inositol prior to membrane isolation; in the other we used exogenous [3H]PIP2 with phosphatidylethanolamine (1:10) added as liposomes to freshly isolated membranes. A Ca2+-dependent PIP2 and PIP phospholipase C activity was characterized by measuring the rate of formation of inositol tris- and bisphosphate. Basal phospholipase C activity was stimulated up to 3-fold by GTP or GTP-gamma-S. Of the two mitogens, alpha-thrombin and EGF, known to stimulate DNA synthesis in Chinese hamster fibroblasts, only alpha-thrombin is a potent activator of PIP2 breakdown in intact cells. Consistent with this observation, alpha-thrombin but not EGF potentiated GTP-gamma-S-dependent phospholipase C activity in membrane preparations. These results strongly support the hypothesis that a GTP-binding protein couples alpha-thrombin receptor to PIP2 hydrolysis. Because both methods used to assay phospholipase C gave identical results, we conclude that the coupling is at the level of PIP2-phosphodiesterase activity. PMID- 3026839 TI - Irreversible inactivation of calcium-dependent proteinases from rat liver by biological disulfides. AB - The effect of low-molecular-mass biological disulfides and their related reduced compounds on the activity of two calcium-dependent neutral proteinases (calpains) from rat liver has been investigated. L-Cystine and L-cystamine bring about the inactivation of both enzymes, while the related reduced compounds L-cysteine and L-cysteamine are without effect. Calpain II is more sensitive to the inactivating effect of glutathione disulfide in comparison with calpain I. The inactivation rates of both calpains depend on the concentration of glutathione disulfide. Reduced glutathione, added at physiological concentration (5 mM), neither affects the proteinase activities nor protects the enzymes from the inactivating effect of glutathione disulfide. The enzymes inactivated by biological disulfides cannot be restored by a large excess of a reducing thiolic compound (dithiothreitol). It is suggested that calcium-dependent proteinases might be inactivated also in vivo by enhanced level of glutathione disulfide. PMID- 3026840 TI - CD4 receptor binding peptides that block HIV infectivity cause human monocyte chemotaxis. Relationship to vasoactive intestinal polypeptide. AB - The octapeptide Ala-Ser-Thr-Thr-Thr-Asn-Tyr-Thr (peptide T) and two structural analogs are potent agonists of human monocyte chemotaxis, evincing identical rank potency orders as was previously shown for their inhibition of human immunodeficiency virus (HIV) envelope binding and T cell infectivity. Chemotactic activity could be inhibited by anti-CD4 monoclonal antibodies (Mabs), but not other mononuclear cell Mabs. The core peptide required for chemotactic activity is a pentapeptide related to the sequence Thr-Thr-Asn-Tyr-Thr. Homologous pentapeptides, identified by computer search, were detected in several other non HIV-related viruses as well as the neuropeptide vasoactive intestinal polypeptide (VIP). The CD4 molecule, therefore, appears to be a recognition molecule for a small signal peptide ligand whose active sequence is a homolog of peptide T and which may be the neuropeptide VIP. PMID- 3026842 TI - Functional expression of cloned cDNA encoding the alpha-subunit of adenylate cyclase-stimulating G-protein. AB - Cloned cDNA encoding the alpha-subunit of the adenylate cyclase-stimulating G protein (Gs), carried by a simian virus 40 vector, has been introduced into the cyc- variant of S49 lymphoma cells by electroporation. In contrast to untransfected cys- cells, clones transformed with the cDNA exhibit an increase in intracellular cyclic AMP concentration in response to a beta-adrenergic agonist. PMID- 3026841 TI - Loss of transforming activity of plasmid DNA (pBR322) in E. coli caused by singlet molecular oxygen. AB - Plasmid DNA pBR322 in aqueous solution was exposed to singlet molecular oxygen (1O2) generated by microwave discharge. DNA damage was detected as loss of transforming activity of pBR322 in E. coli (CMK) dependent on the time of exposure. DNA damage was effectively decreased by singlet-oxygen quenchers such as sodium azide and methionine. Replacement of water in the incubation buffer by D2O led to an increase in DNA damage. 9,10-Bis(2-ethylene)anthracene disulfate was used as a chemical trap for 1O2 quantitation by HPLC analysis of the endoperoxide formed. PMID- 3026843 TI - Structure of the porin from a bacterial stalk. AB - The stalks (hyphae) of a prosthecate bacterium, directly sampled from the water surface of a hot pond, show extended regular patterns on their envelope in the electron microscope. Image processing revealed a structure of the crystalline complexes which is very similar to the gross morphology of the Escherichia coli porins OmpC and OmpF. The natural two-dimensional crystal of the outer membrane protein has p3 symmetry and a lattice constant of 7.95 nm. The three-dimensional structure of the stalk porin has been determined to an almost isotropic resolution of 1.7 nm. The reconstruction revealed a complex network of channels within the membrane matrix with a triplet of pores merging into a common outlet, similar to the structure of the E. coli porin OmpF in reconstituted membranes. In addition, a blindly ending pore exists which appears to be connected to the continuous pores via small channels. The significance of the regularly arrayed porin cylinders with respect to the shape and function of the stalks is discussed. PMID- 3026844 TI - Sulfhydryl groups are involved in H+ translocation via the uncoupling protein of brown adipose tissue mitochondria. AB - Mersalyl inhibits H+ transport via the uncoupling protein (UP) in brown adipose tissue (BAT) mitochondria estimated as swelling in potassium acetate (Ki 67 microM) or as valinomycin-induced H+ extrusion in K2SO4 (Ki 55 microM) and KCl. The swelling in KCl is depressed only slightly. Some other SH-reagents (p hydroxymercuribenzoate, 5,5'-dithiobis(2-nitrobenzoate) and thiolyte DB), but not hydrophobic reagents (N-ethylmaleimide and eosin-5-maleimide), exhibit analogous inhibition. Thus an essential SH-group localized at the water-accessible cytosolic surface of UP was found to be involved in H+ transport via UP but not in Cl- transport. PMID- 3026845 TI - Regulation of adenylate cyclase by hormones and G-proteins. AB - Over the past few years, it has become apparent that a large number of transmembrane signaling systems operate through heterotrimeric G-proteins [( 1] Gilman, A.G. (1984) Cell 36, 577-579; [2] Baker, P.F. (1986) Nature 320, 395). Adenylate cyclase is regulated by stimulatory hormones through Gs(alpha s beta gamma) and inhibitory hormones through Gi(alpha i beta gamma) [( 2]; Katada, T. et al. (1984) J. Biol. Chem. 259, 3586-3595), whereas the breakdown of phosphatidylinositol bisphosphate (PIP2) to inositol trisphosphate (IP3) and diacylglycerol (DG) by phospholipase C is probably also mediated by a heterotrimeric G-protein (Go or Gi) [1,2]. Similarly, the activation of cGMP phosphodiesterase by light-activated rhodopsin is mediated through the heterotrimeric G-protein transducin (Stryer, L. (1986) Rev. Neurosci. 9, 89-119). Other transmembrane signaling systems may also be found to involve G-proteins similar to those already recognized. Because of the emerging universality of G proteins as transducers of receptor-triggered signals, it may be useful to evaluate the current models prevailing in the adenylate cyclase field, as these models seem to guide our way in evaluating the role of G-proteins in transmembrane signaling, in general. PMID- 3026846 TI - Chromogranin A can act as a reversible processing enzyme inhibitor. Evidence from the inhibition of the IRCM-serine protease 1 cleavage of pro-enkephalin and ACTH at pairs of basic amino acids. AB - Bovine parathyroid chromogranin A inhibits the cleavage of Z-Ala-Lys-Arg-AMC by either trypsin or IRCM-serine protease 1 (IRCM-SP1), a putative novel processing enzyme originally isolated from porcine pituitary anterior and neurointermediate lobes. On larger substrates, chromogranin A is a reversible competitive inhibitor of the cleavage at pairs of basic amino acids by IRCM-SP1. The substrates tested included pituitary ACTH and adrenal medulla pro-enkephalin-derived peptides such as the 8.6 kDa synenkephalin-containing precursor and peptide B. Chromogranin A is itself selectively processed by IRCM-SP1, and ACTH was shown to compete for such cleavage. These data suggest that chromogranins as a class of acidic proteins could participate in the tissue-specific processing of pro-hormones. PMID- 3026847 TI - Phorbol ester induces loss of VIP stimulation of adenylate cyclase and VIP binding sites in HT29 cells. AB - Treatment of HT29 cells with the tumor promoting phorbol ester PMA resulted in an attenuation of VIP-stimulated cAMP production in intact cells and VIP-stimulated adenylate cyclase activity in cell membranes. PMA did not decrease the ability of cholera toxin and forskolin to elevate cAMP levels in intact cells. Fluoride stimulated adenylate cyclase activity in HT29 cells homogenates was not affected by PMA. The maximal VIP binding capacity of homogenates prepared from HT29 cells treated with PMA was decreased by 50%. It is concluded that protein kinase C regulates VIP receptor function possibly through phosphorylation of the VIP receptor. PMID- 3026848 TI - Cloning and sequence analysis of the human brain beta-adrenergic receptor. Evolutionary relationship to rodent and avian beta-receptors and porcine muscarinic receptors. AB - Two cDNA clones, lambda-CLFV-108 and lambda-CLFV-119, encoding for the beta adrenergic receptor, have been isolated from a human brain stem cDNA library. One human genomic clone, LCV-517 (20 kb), was characterized by restriction mapping and partial sequencing. The human brain beta-receptor consists of 413 amino acids with a calculated Mr of 46480. The gene contains three potential glucocorticoid receptor-binding sites. The beta-receptor expressed in human brain was homology with rodent (88%) and avian (52%) beta-receptors and with porcine muscarinic cholinergic receptors (31%), supporting our proposal [(1984) Proc. Natl. Acad. Sci. USA 81, 272 276] that adrenergic and muscarinic cholinergic receptors are structurally related. This represents the first cloning of a neurotransmitter receptor gene from human brain. PMID- 3026849 TI - Solute carriers involved in energy transfer of mitochondria form a homologous protein family. AB - The sequences of three mitochondrial carriers involved in energy transfer, the ADP/ATP carrier, phosphate carrier and uncoupling carrier, are analyzed. Similarly to what has been previously reported for the ADP/ATP carrier and the uncoupling protein, now also the phosphate carrier is found to have a tripartite structure comprising three similar repeats of approx. 100 residues each. The three sequences show a fair overall homology with each other. More significant homologies are found by comparing the repeats within and between the carriers in a scheme where the sequences are spliced into repeats, which are arranged for maximum homology by allowing possible insertions or deletions. A striking conservation of critical residues, glycine, proline, of charged and of aromatic residues is found throughout all nine repeats. This is indicative of a similar structural principle in the repeats. Hydropathy profiles of the three proteins and a search for amphipathic alpha-spans reveal six membrane-spanning segments for each carrier, providing further support for the basic structural identity of the repeats. The proposed folding pattern of the carriers in the membrane is exemplified with the phosphate carrier. A possible tertiary arrangement of the repeats and the membrane-spanning helices is shown. The emergence of a mitochondrial carrier family by triplication and by divergent evolution from a common gene of about 100 residues is discussed. PMID- 3026851 TI - Agonist-induced alpha 1-adrenergic receptor changes. Evidence for receptor sequestration. AB - Short-term receptor regulation by agonists is a well-known phenomenon for a number of receptors but little is known about the regulation of alpha 1 adrenergic receptors. In the present study we provide evidence of alpha 1 adrenergic receptor changes induced by agonists on DDT1 MF-2 smooth muscle cells. The cells were preincubated with the agonist and receptor changes were investigated in the cells washed free of the agonist. On cells pretreated with norepinephrine the number of receptors recognized by [3H]prazosin at 4 degrees C was reduced by 38%. The receptors were not degraded as the number of sites was the same in control and norepinephrine-treated cells when binding was measured at 37 degrees C. When binding was measured on fragmented membranes (at 4 degrees C), the number of receptors was the same in control and norepinephrine-treated cells, suggesting that the disruption of cellular integrity might expose receptors which are probably sequestered after agonist treatment. We conclude that agonists induced rapid sequestration of receptors on intact DDT cells. PMID- 3026850 TI - Inhibition of Na+/H+ exchange reduces Ca2+ mobilization without affecting the initial cleavage of phosphatidylinositol 4,5-bisphosphate in thrombin-stimulated platelets. AB - Stimulation of human platelets increases cytoplasmic pH (pHi) via activation of Na+/H+ exchange. We have determined the effect of inhibiting Na+/H+ exchange on (i) thrombin-induced Ca2+ mobilization and (ii) turnover of 32P-labelled phospholipids. Blocking Na+/H+ exchange by removal of extracellular Na+ or by ethylisopropylamiloride (EIPA) inhibited Ca2+ mobilization induced by 0.2 U/ml thrombin, whereas increasing pHi by NH4Cl enhanced the thrombin-induced increase in cytosolic free Ca2+. The effect of EIPA was bypassed after increasing pHi by moneasin. The thrombin-induced cleavage of phosphatidylinositol 4,5-bisphosphate (PIP2) was unaffected by treatments that blocked Na+/H+ exchange or increased pHi. It is concluded that activation of Na+/H+ exchange is a prerequisite for Ca2+ mobilization in human platelets but not for the stimulus-induced hydrolysis of PIP2. PMID- 3026852 TI - Brush border myosin heavy chain phosphorylation is regulated by calcium and calmodulin. AB - Myosin from chicken intestinal brush borders is phosphorylated on its heavy chains at threonine by a kinase isolated from brush borders. In contrast to other heavy chain kinases, the brush border kinase activity is dependent on calcium and calmodulin. The partially purified preparation also phosphorylated myosin on its light chains at serine, but in a calmodulin-independent manner. Phosphorylation of the light chains in the absence of calmodulin or both heavy and light chains in the presence of calmodulin activated its actin-activated ATPase activity about 10-fold, to about 50 nmol/min per mg. PMID- 3026853 TI - Ribose-5-phosphate isomerase and ribulose-5-phosphate kinase show apparent specificity for a specific ribulose 5-phosphate species. AB - Ribose-5-phosphate isomerase and ribulose-5-phosphate kinase appear to show specificity for a particular ribulose 5-phosphate species. The effect of this specificity will be channeling of ribulose 5-phosphate from the isomerase to the kinase during photosynthesis. PMID- 3026854 TI - Slugs and snails and opiate tales: opioids and feeding behavior in invertebrates. AB - There is accumulating evidence that opioid systems are involved in the regulation of fundamental behavioral and physiological processes in invertebrates. Feeding is a basic physiological function that is essential for maintaining homeostasis. Results of studies examining the feeding responses of molluscs and arthropods treated with various opiate agonists and antagonists indicate that delta, kappa, mu, and possibly sigma opioid systems differentially and selectively mediate the components of their natural feeding behavior. Moreover, it appears that at an early evolutionary stage the mu and kappa systems have developed to selectively affect the components of feeding behavior associated with the acquisition and ingestion of food. In addition, evidence suggests that neuropeptides that have been proposed as possible endogenous antagonists of opioid-mediated feeding in mammals may also be involved in the control of feeding in invertebrates. This indicates that there may be an interplay of opioid agonists and antagonists in the regulation of feeding and satiation in invertebrates analogous to that proposed for vertebrates. Moreover, these findings indicate that opioid influences on feeding have been conserved through evolution. PMID- 3026855 TI - Opioid peptides and the control of feeding in sheep. AB - Opioid peptides, particularly beta-endorphin, methionine- (MEK) and leucine enkephalin, and dynorphin, are involved in the regulation of food intake in mammals. The precursor molecules of these peptides undergo differential processing in brain areas producing regional concentration differences in opioids. Intraregional concentration changes also accompany alterations in feeding states. For example, MEK concentrations decrease in the basomedial hypothalamus, amygdala, and olfactory bulb in fed sheep compared with fasted sheep. Moreover, these changes are species specific. In sheep, beta-endorphin decreases in the dorsomedial and posterior hypothalami after feeding, but in the rat it is increased in the ventromedial hypothalamus and decreased in the posterior hypothalamus. In addition, immunohistochemical localization of cell bodies shows interspecies differences in concentrations. For example, dynorphin is found predominantly in the suprachiasmatic area in sheep, but in the paraventricular nucleus in the rat. These observations indicate that regulation of food intake may be differentially controlled in these species. In sheep, kappa agonists increase food intake, whereas stimulation of delta receptors inhibits feeding. Further clarification of the receptors involved in food intake will necessitate studies with more specific agonists. PMID- 3026856 TI - Biochemical characteristics of receptors for vasoactive intestinal polypeptide in nervous, endocrine, and immune systems. AB - Neuropeptides have recently been shown to modulate the immune response. Vasoactive intestinal polypeptide (VIP) modulates lymphocyte migration to Peyer's patches and inhibits natural killer cell activity. VIP receptors have been characterized on lymphocytes and have been compared with the VIP receptor in nervous tissue and in tissues of the gastrointestinal tract. The evidence supports the existence of at least two classes of high-affinity VIP receptors as well as a low-affinity receptor. The development of tissue-specific agonists and antagonists to the VIP receptor may thus be feasible. PMID- 3026857 TI - Molecular and cellular properties of human polymorphonuclear leukocyte receptors for leukotriene B4. AB - The distinctive characteristics of human polymorphonuclear (PMN) leukocyte receptors for leukotriene B4 (LTB4) have been elucidated by studies of binding of [3H]LTB4, the structure of protein constituents of the receptors isolated from plasma membranes, and the effects of antireceptor antibodies. A high-affinity class of 4400 receptors with a KD of 0.4 nM mediates chemotaxis and increased adherence of PMN leukocytes, whereas a low-affinity class of 270,000 receptors with a KD of 61 nM mediates the release of lysosomal enzymes and increases in oxidative metabolism. The low-affinity receptors are composed of a 60,000-dalton protein-binding unit. The high-affinity receptors are composed of the same binding unit in association with a 40,000-dalton guanine nucleotide-binding protein. That antireceptor antibodies as well as LTB4 distinguish the two classes of receptors with different functional consequences suggests the possibility of unique approaches to the regulation of leukocyte function at the receptor level. PMID- 3026858 TI - Binding of leukotriene C4 by glutathione transferase: a reassessment of biochemical and functional criteria for leukotriene receptors. AB - Studies of the molecular structures and biological activities of leukotrienes (LTs) provided evidence for the presence of multiple, subclass-specific receptors on the surface of responding tissues. Distinct receptors for LTC4 and LTD4 have been defined based on functional and metabolic criteria. However, radioligand binding studies of LTC4-binding sites revealed anomalous results that failed to demonstrate a parallel relationship between binding affinity and functional activity for a number of agonists. In this study, we identified a high-affinity binding unit for LTC4 as the Ya subunit containing glutathione transferases (EC 2.5.1.18) that is present in both the cytosolic and the membrane fractions of rat liver homogenate. This enzyme accounted for a substantial portion of the LTC4 binding activity in rat liver cytosol as well as in mitochondrial and microsomal fractions. We suggest that the LTC4-binding sites in tissues are heterogeneous and that some binding units may have functions other than transduction of a signal across the cell membrane. PMID- 3026859 TI - Functional characteristics of neuropeptides in the dorsal medulla oblongata and vagus nerve. AB - The dorsomedial medulla plays an integral role in the processing of primary sensory afferent information from the respiratory, cardiovascular, and gastrointestinal systems. A correlation has also been made between the topographical organization of these vagal afferent fibers in the dorsal medulla and the distribution of a variety of neuropeptides and their receptors in this brain region. In this paper, the evidence for the presence of several neuropeptides and their receptors in the dorsomedial medulla and intra- and/or extracranial segments of the vagus nerve is presented. The possible physiological significance of these peptides and their putative receptors in the vagus nerve is also addressed, with emphasis on angiotensin II and its cardiovascular actions in this region. PMID- 3026861 TI - Aquaculture: still promising. PMID- 3026860 TI - Reflex regulation of the circulation after stimulation of cardiac receptors by prostaglandins. AB - Prostaglandins (PGs) are potent vasoactive substances that may participate in the control of coronary blood flow, platelet aggregation, and inflammation. An important action of PGs may be the stimulation of c fibers in general and vagal cardiac c fibers in particular. The Bezold-Jarisch reflex after intracoronary injection of Veratrum alkaloids is very similar to the vagal bradycardia elicited by stimulation of cardiac PG synthesis or injection of prostacyclin (PGI2). The characteristic features of this reflex are 1) stimulation of c fibers, 2) inferoposterior wall location of receptors, 3) vagal afferents, 4) vagal efferents to the heart, 5) sympathetic efferents to peripheral blood vessels, and 6) interaction with other reflexes. Vagal cardiac c fibers are activated by intracoronary injections of PGI2 or arachidonic acid, resulting in a vagal reflex bradycardia and hypotension due to withdrawal of peripheral alpha-adrenergic tone to resistance vessels. The cardiac receptors are located predominantly in the inferoposterior wall of the left ventricle. When stimulated by PGs, cardiac receptors may also modify the regulation of arterial pressure by the baroreflexes, altering the inverse relationship between systemic arterial pressure and heart rate. Thus, there is a striking parallelism between the veratridine-induced Bezold-Jarisch reflex and PG-induced cardiac reflexes, although the physiological and clinical significance of these reflexes remains to be determined. PMID- 3026863 TI - Studies into disturbing receptor 'memory' in a unicellular (Tetrahymena) model system: changes in the imprinting potential on exposure to combinations of related and unrelated hormones. AB - Signal molecules (hormones) can induce a hormonal imprinting in the unicellular Tetrahymena, as judged from an increase in binding capacity on reexposure in the subsequent generations. Structurally unrelated polypeptide hormones (insulin, FSH) neutralize each other's effect, whereas related hormones (TSH-FSH) may amplify or depress it. The non-signal polypeptide molecule BSA prevents hormonal imprinting on combined exposure. The glucocorticoid hormone enhances binding capacity for insulin (as in mammals) and increases its imprinting potential, whereas on TSH and FSH it has no similar effects. The amino acid hormone serotonin increases the binding capacity for TSH and FSH in non-pretreated cells, but depresses rather than stimulates the imprinting potential of these hormones, measured in the case of the second encounter. PMID- 3026862 TI - Retinoic acid-induced differentiation of human neuroblastoma: a cell variant system showing two distinct responses. AB - Retinoic acid (RA) has been shown to induce the differentiation of human neuroblastoma cells in vitro. In this study, we describe two variants of the SK-N SH human neuroblastoma cell line that have dramatically different responses to RA. RA induces neuronal-like differentiation characterized by extensive neurite outgrowth, thick neurite bundles, and large cellular aggregates of SK-N-SH-N (SH N) cells. In contrast, RA treatment of SK-N-SH-F (SH-F) cultures transforms the small neuroblast cells into large flattened, fibroblastic or epithelial-like cells. Karyotype analysis verified that the SH-N and SH-F cultures were derived from a common precursor cell. Confirmation of their markedly different responses to RA was obtained by metabolic labelling of glycoproteins and SDS-PAGE analysis. While both sublines showed very similar Coomassie-labelled protein bands and glycoprotein profiles in control cultures, dramatic differences between the lines were revealed following RA treatment. In contrast to their similar protein profiles, untreated SH-N and SH-F cells had quite different patterns of ganglioside biosynthesis in that GM3 was detected in SH-F cells but not in SH-N, while GM1 was only detected in SH-N. Cellular RA binding protein (CRABP) was detected in both SH-F and SH-N cells and their RA-transformed derivatives. These results demonstrate heterogeneity in the response to RA of neuroblastoma cells derived from a common origin that cannot be accounted for by differences in CRABP content. The SH-N and SH-F neuroblastoma sublines should provide a useful system for further studies of the molecular processes through which RA exerts its differentiation-inducing activity on this type of tumor. PMID- 3026864 TI - Transbilayer distribution of phospholipid synthesizing enzymes in the sarcoplasmic reticulum membrane. AB - The topology of enzymes involved in phospholipid synthesis in the membrane of sarcoplasmic reticulum vesicles was investigated by means of limited proteolysis. The digestion of sarcoplasmic reticulum proteins with different proteases caused the considerable decrease of the activities of choline- and ethanolaminephosphotransferases without significant inactivation of glycerophosphate and lysophosphatidylcholine acyltransferases. On the basis of these results we concluded that the active sites of phosphotransferases are located on the cytoplasmic side, whereas the active sites of acyltransferase are present either on the extracytoplasmic side or burried into the sarcoplasmic reticulum membrane. PMID- 3026865 TI - Effects of gamma-linolenic acid supplementation on lipogenesis regulation in pregnant zinc-deficient rat and fetus. AB - In the liver of zinc-deficient pregnant rats fatty acid synthetase and delta 9 desaturase activities decreased and diet supplementation with gamma-linolenic acid potentiated this effect. However, in liver microsomes from the foetuses of zinc-deficient mothers, HMG CoA reductase and delta 9-desaturase activities declined but fatty acid activity rose. The same applied to foetuses from mothers whose diet was supplemented with gamma-linolenic acid and here again, the effect of zinc deficiency was potentiated. The fact that delta 9 activity dropped whereas fatty acid synthetase activity rose implied a defect in the mechanism regulating the functioning of these enzymes. In the non zinc-deficient group of pregnant females, gamma-linolenic acid supplementation had no effect on fatty acid synthetase and delta 9-desaturase activities but significantly increased HMG CoA reductase activity. In foetuses from the same group, the activities of MHG CoA reductase, delta 9-desaturase and fatty acid synthetase all increased. PMID- 3026866 TI - Distribution of the short dispersed repetitive sequences B1 and B2 in the mouse genome. AB - The organization of the short dispersed repetitive sequences B1 and B2 in the mouse genome was investigated by hybridization of randomly selected genomic clones with isolated and labelled in vitro B1 and B2. Cloning and restriction mapping experiments indicated that these two DNA sequences were not entirely independently distributed along mouse DNA, but approximately half of them formed heterologous pairs separated by stretches of apparently random DNA. PMID- 3026867 TI - Activity of deoxyribonucleoside-activated nucleotidase in cells from various lymphoid mouse tissues and in concanavalin A-stimulated lymphocytes. AB - The specific activity in cells from lymph nodes, spleen and thymus was 32, 28 and 25 nmol/min per mg of cytosol protein, respectively, whereas that in bone marrow cells was significantly lower (10 units/mg of protein). No difference in specific DAN activity between isolated B- and T-lymphocytes was observed. Two types of lymphoid mouse cell lines (MOPC-31C plasmacytoma cells, S49 Cyc- lymphoma cells) showed specific activities similar to the normal lymphoid cells. In concanavalin A- stimulated spleen lymphocytes in culture there was a rapid increase in DAN activity shortly after maximum DNA synthesis, reaching a plateau 2-3 times the original level. The enzyme (DAN) of mouse tissues possessed the characteristic properties previously detected for the rat enzyme. PMID- 3026868 TI - Preparation and characterization of submitochondrial fractions from adrenal cells. AB - A method for preparing submitochondrial fractions from adrenocortical cells was developed by adapting a procedure that has been successful with yeast mitochondria. The method is based upon osmotic swelling, sonication and centrifugation in sucrose. The preparation yields highly purified fractions of outer membrane, intermembrane space, inner membrane and a less purified fraction of matrix. Recoveries are good so that 10(7) cells yield approximately 170 micrograms of inner membrane protein and 12 micrograms of outer membrane protein. Electron microscopy shows that the outer membrane fraction consists of vesicles (0.2-0.6 micron diameter) while inner membrane appears as densely packed sheets of membranous material. Two-dimensional polyacrylamide gels (isoelectric focusing followed by electrophoresis) of all the fractions give highly reproducible patterns of protein spots with Coomassie staining. Steroidogenic proteins were found only in inner membrane fractions which were shown to contain cytochrome P 450 C27 side-chain cleavage and P-450 11 beta-hydroxylase together with adrenodoxin and adrenodoxin reductase. Inner membrane catalyzes side-chain cleavage of cholesterol (conversions to pregnenolone) and 11 beta-hydroxylation (DOC----corticosterone) when substrate and NADPH are added. The preparation yields highly purified submitochondrial fractions from Y-1 mouse adrenal tumor cells and from porcine and bovine adrenocortical mitochondria. The method has the virtue of yielding highly purified intermembrane fluid which is not true of other methods for fractionation of adrenal mitochondria. The procedure also yields cleaner preparations of the two membranes than two other published methods currently used to prepare submitochondrial fractions from adrenocortical cells. PMID- 3026869 TI - Regulation of crab Y-organ steroidogenesis in vitro: evidence that ecdysteroid production increases through activation of cAMP-phosphodiesterase by calcium calmodulin. AB - In decapod crustaceans steroidogenic glands (Y-organs) produce the molting hormone, ecdysone. A putative neuropeptide, molt-inhibiting hormone (MIH), released from eyestalk neurosecretory cells, directly regulates Y-organs by suppressing steroidogenesis; the effect is mediated by an increase in cAMP. We explored calcium-cAMP interactions in the regulation of Y-organs in vitro of the crab, Cancer antennarius. Basal ecdysteroid production was enhanced by extracellular calcium (EC). MIH suppression did not require EC but its action was blocked by high EC. The inhibitors of Ca2+ flux, lanthanum and ruthenium red, mimicked and enhanced MIH action. Calcium ionophore A23187 raised basal steroidogenesis dose-dependently (10(-6) to 10(-4) M) and with time course (effect evident after 2 h) similar to that of suppression by MIH. Low EC enhanced the suppressive effects on steroidogenesis of forskolin and dibutyryl cyclic AMP ((Bu)2cAMP) but not of MIH, lysine vasopressin (LVP), or 3-isobutyl-1-methyl xanthine (IBMX); basal Y-organ cAMP levels were elevated by low EC and reduced by A23187. A23187 reduced the steroidogenic-suppressive effects of MIH, LVP, forskolin and (Bu)2cAMP but not of IBMX; rises in cAMP induced by MIH, LVP, and forskolin but not by IBMX were blunted by A23187. These findings suggested a stimulatory action of calcium on phosphodiesterase (PDE). The calmodulin (CM) inhibitor trifluoperazine (TFP; 10(-5) to 10(-4) M) reduced basal and A23187 stimulated steroidogenesis, enhanced the inhibitory effects of MIH and (Bu)2cAMP on ecdysteroid production, enhanced the stimulatory effects of MIH and forskolin on cAMP, and blocked the inhibition of cAMP by A23187. Y-organ PDE activity was enhanced by increasing free Ca2+ (10(-7) to 10(-5) M) and inhibited by TFP (10( 5) to 10(-4) M). Adenylate cyclase activity of Y-organ cell particulate fraction was unaffected by Ca2+ or TFP. Calcium stimulates steroidogenesis, apparently by activating a calcium-CM-dependent cAMP-PDE: the action is counter to the cAMP mediated MIH-inhibitory system. Ca2+ fluxes were measured with dispersed Y-organ cells, in the presence and absence of agents that alter cAMP levels. The ionophore A23187, but not MIH or forskolin, increased 45Ca2+ entry by 45% over untreated control cells. Efflux from 45Ca2+-preloaded cells was increased 30% by MIH and forskolin, but not A23187. These data, together with those further above, suggest that MIH suppresses steroidogenesis in part by fostering Ca2+ depletion, and that the effect is mediated by cAMP. PMID- 3026870 TI - Octopamine and cyclic AMP mediate release of adipokinetic hormone I and II from isolated locust neuroendocrine tissue. AB - Octopamine serves as a neurotransmitter in the glandular lobe of the locust corpus cardiacum where it regulates adipokinetic hormone (AKH) secretion from intrinsic neurosecretory cells. Two AKHs (AKH I and II) from the corpus cardiacum of Locusta have been sequenced and synthesized. We have now demonstrated that octopamine mediates release of both AKH I and II from Locusta corpora cardiaca in vitro. Octopamine, IBMX, and forskolin have previously been shown to elevate levels of cAMP in the glandular lobe. In this paper we demonstrate that IBMX and forskolin mediate secretion of AKH I and AKH II thus mimicking the effects of octopamine in this tissue. The cAMP analogs dibutyryl cAMP and 8-bromo cAMP also elicit release of AKH and a subthreshold concentration of IBMX potentiates the effects of octopamine. These observations demonstrate that cAMP participates in regulating AKH release and support the hypothesis that octopamine mediates hormone release at least in part via changes in intracellular levels of cAMP. The release of AKH I and AKH II is apparently regulated by similar mechanisms. The hyperlipemic activity of AKH II released into the perfusates following stimulation by octopamine and agents which elevate cAMP levels is significantly lower than that of AKH I. This differential response is probably due both to the reduced lipid-mobilizing effect of AKH II relative to AKH I, as well as to the release of greater amounts of AKH I. PMID- 3026871 TI - Dissociation between pituitary GnRH binding sites and LH response to GnRH in vitro. AB - Experiments were performed to study gonadotroph responsiveness to gonadotrophin releasing hormone (GnRH) in vitro in dispersed pituitary cells from ovariectomised rats and mice when GnRH binding sites were increased or reduced, respectively. Maximal/basal LH release after GnRH treatment of intact female rat pituitary cells was 4.7 to 9.7-fold (range n = 3 expts.) compared to 3.4 to 5.0 fold for cells from ovariectomised rat donors. Both basal and maximal GnRH stimulated LH release from ovariectomised (OVX) rat pituitary cells were 1.5 to 3 fold greater than from intact rat cells, which corresponded to increased LH content of the cells. There was no significant change in the GnRH ED50 concentration (intact = 2.3 +/- 0.03 X 10(-10) M; OVX = 3.3 +/- 0.08 X 10(-10) M (mean +/- SEM, n = 3 expts.)), despite a 57-88% increase in GnRH binding sites in ovariectomised rat pituitary cells. Basal and maximal LH release from ovariectomised mouse pituitary cells was 1.5 to 3-fold greater than that from intact mouse pituitary cells. There was no change in the GnRH ED50 concentration (intact = 4.3 +/- 2.3 X 10(-9) M; OVX = 3.4 +/- 0.9 X 10(-9) M (mean +/- SEM, n = 3 expts.)), even though GnRH binding sites were reduced by 40-73% in the cells from ovariectomised mice. These data indicate that changes in GnRH binding sites of the magnitude observed after ovariectomy play no part in the regulation of gonadotroph responsiveness to GnRH, which is determined by changes in post receptor events, one of which is an increase in cellular LH. PMID- 3026872 TI - Human growth hormone-stimulated mitogenesis of Nb2 node lymphoma cells is not mediated by an immediate acceleration of phosphoinositide metabolism. AB - Lactogenic hormones stimulate the mitogenesis of Nb2-11C rat lymphoma cells and this stimulation is enhanced by 12-O-tetradecanoyl phorbol ester (TPA). The effect of human growth hormone (hGH) and TPA on phosphoinositide metabolism in Nb2-11C cells was tested by measuring the incorporation of [3H]myo-inositol or 32P or measuring the breakdown of polyphosphoinositides in cells prelabeled with [3H]myo-inositol, 32P or [3H]arachidonic acid. We found that none of these processes is immediately stimulated by either hGH and/or TPA. Our results indicate that the overall phosphoinositide turnover is slow or confined to small fractions only. On the other hand, the phosphorylation-dephosphorylation cycle of polyphosphoinositides is quite fast and remarkably exceeds the rate of incorporation of [3H]myo-inositol. Our present results indicate, therefore, that the mitogenic effect of hGH and its enhancement by TPA is not mediated by an immediate increase in phosphoinositide metabolism. PMID- 3026873 TI - Mechanism of action of prostaglandin F2 alpha-induced luteolysis: evidence for a rapid effect on the guanine nucleotide binding regulatory component of adenylate cyclase in rat luteal tissue. AB - The initial events in prostaglandin F2 alpha-(PGF2 alpha)-induced luteolysis were studied in pregnant mare serum gonadotropin/human chorionic gonadotropin (PMSG/hCG)-treated rats with luteinized ovaries. Injection with a potent PGF2 alpha analog (cloprostenol, 5 micrograms/ml) induced functional luteolysis, as assessed by plasma levels of progesterone and 20 alpha-dihydroprogesterone. At 0.5 and 3 h after cloprostenol administration the luteolytic effect was also evident as a reduced response of luteal adenylate cyclase to all stimulatory agents tested, LH, isoproterenol, fluoride, guanylylimidodiphosphate and forskolin. 24 h after cloprostenol the response to all agents, except to LH, had returned to normal. This general and transient block of the luteal adenylate cyclase system indicates that a common factor, possibly the stimulatory guanine nucleotide binding protein (Ns), is involved in the mechanism of action of PGF2 alpha. To test this hypothesis, we measured the functional coupling of the Ns protein to the beta-adrenergic receptor in luteal membranes. Binding competition curves showed a marked shift to the right in membranes prepared from rats injected with cloprostenol 0.5 and 3 h before membrane preparation, while at 24 h after cloprostenol the shift had disappeared. The total number of beta-adrenergic receptors was, however, not affected by the cloprostenol treatment. Computer analysis of the data indicates that, at 0.5 and 3 h after cloprostenol treatment, there was a reduced number of high affinity binding sites, 38 and 41%, respectively, compared to 53% for control membranes. The cellular mechanism for this action of PGF2 alpha on the Ns protein remains to be elucidated. PMID- 3026874 TI - Melanocyte-stimulating hormone affects melanogenic differentiation of quail neural crest cells in vitro. AB - Quail neural crest cells were treated in vitro with alpha-melanocyte-stimulating hormone (alpha-MSH) or dibutyryl cyclic AMP (dbcAMP) plus theophylline. These treatments increased the proportion of melanocytes to total cells in crest cell outgrowth cultures. Pigmentation of neural crest cell clusters proceeded more rapidly when cultures were treated with alpha-MSH or dbcAMP plus theophylline than when untreated. In clonal cell cultures, the proportion of pigmented colonies to total colonies was increased by MSH treatment. From these results, MSH seems not only to accelerate melanogenic differentiation but also to affect the state of commitment of neural crest cells to melanogenic differentiation in vitro, and this action of MSH appears to be mediated by cAMP. PMID- 3026875 TI - Repeated failure of prenatal ACTH administration to alter masculine behavior in mice. AB - In four experiments, different preparations and modes of prenatal administration of ACTH all failed to produce any substantial effects upon male sexual behavior in mice. In Experiment 1, CD-1 females implanted during the third trimester of pregnancy with osmotic pumps releasing varied dosages of ACTH1-24 produced male offspring with essentially normal copulatory behavior. In Experiment 2, prenatal injections of high doses of ACTH1-24 had no effect upon male sexual activity. In Experiment 3, osmotic pumps releasing ACTH1-39 during the third trimester of pregnancy had no effect upon sexual behavior of offspring. However, aggressive behavior was significantly reduced, relative to untreated controls, in offspring of all females implanted with pumps, including those releasing only saline. In Experiment 4, third-trimester injections of ACTH1-39 in long-acting gel form had no effect on the sexual behavior or aggression of offspring of C57 strain females. In most of these experiments, ACTH treatment significantly reduced body weight. These results do not confirm previous suggestions that pituitary-adrenal hormones influence the perinatal differentiation of sexually dimorphic behavior. PMID- 3026877 TI - Banting lecture 1986. Does a common mechanism induce the diverse complications of diabetes? PMID- 3026876 TI - Inhibitor of calmodulin and cAMP phosphodiesterase activity in BB rats. AB - Diabetes mellitus in humans is associated with increased plasma and tissue levels of cAMP and decreased cAMP phosphodiesterase (PDE) activity. Calmodulin (CM) is a low-molecular-weight protein essential for activation of cAMP PDE. The inhibitor (INH) is a low-molecular-weight substance that inhibits the activity of CM in multiple systems, including PDE. Spontaneously diabetic BB rats (SDR) and their nondiabetic littermates (NDR) were used in this study. Holtzman rats were rendered diabetic by streptozocin (STZ). STZ-induced diabetic rats (STZ-DR) and BB rats were studied with and without the benefit of insulin therapy. Calmodulin was assayed both by bioassay and by specific radioimmunoassay. The inhibitor was bioassayed by its ability to inhibit CM-activated PDE. Results showed that both spontaneous and STZ-induced diabetes are associated with a decrease in activity of the low-Michaelis constant (Km) cAMP PDE in the liver (39%, SDR; 70% STZ-DR). Calmodulin activity was also decreased in the livers of both animals (13%, SDR; 68%, STZ-DR). Similar data were obtained for NDRs. The inhibitor, on the other hand, was increased in the livers of untreated SDRs and STZ-DRs (155%, SDR; 125%, STZ-DR). No change was noted for NDRs. All these changes were restored toward normal after treatment with insulin. These data suggest that in diabetes the defect in the cAMP PDE-CM-INH system is demonstrated in both an environmental model, as illustrated by STZ-DRs, and a genetic model, as shown by SDRs and NDRs. The inhibitor activity, however, is not changed significantly in NDRs. We speculate that the inhibitor activity plays a role in dictating whether the genetic NDR will or will not become clinically diabetic. PMID- 3026878 TI - Insulin secretory responses of a clonal cell line of simian virus 40-transformed B cells. AB - We have evaluated the potential of the clonal insulin-secretory cell line HIT-T15 as a model system for investigating stimulus-secretion coupling in pancreatic B cells. In contrast to other cell lines, HIT cell insulin secretion was consistently stimulated 2- to 3-fold by D-glucose. The maximally effective concentration of glucose was 10 mmol/l; between 2 and 10 mmol/l glucose the increase in insulin release was paralleled by an increased rate of glucose oxidation. The main characteristics of glucose-stimulated insulin release by HIT cells were essentially similar to those of normal islets. Thus, the response was specific for metabolizable sugars (D-mannose and D-glyceraldehyde stimulated insulin release but L-glucose and D-galactose were ineffective); markedly dependent on extracellular Ca2+ concentration; potentiated by forskolin, glucagon, acetylcholine and 12-O-tetradecanoyl phorbol 13-acetate; inhibited by adrenaline or somatostatin; showed a biphasic pattern of release in perifusion experiments, with both phases being potentiated by forskolin. The secretory response of the HIT cells to amino acids was also similar to that of normal islets. Thus, L-leucine and its deamination product 2-ketoisocaproate were effective stimuli, whereas L-isoleucine and L-glutamine were ineffective. Insulin release from HIT cells could also be evoked by the sulphonylureas glibenclamide and tolbutamide and by an increase in concentration of extracellular K+ to 40 mmol/l. The content of cyclic AMP in HIT cells was increased modestly by glucose but not by an increase in extracellular K+. Forskolin elicited a 4-fold increase in cyclic AMP content. We conclude that HIT cells retain the essential features of the insulin secretory response of normal B cells and represent an important tool for further biochemical characterization of the secretory system. PMID- 3026880 TI - [Potential malignancy of cystosarcoma phyllodes of the breast. A contribution to the histogenesis of fibrosarcoma]. AB - The present case report, which describes the transformation of a cystosarcoma phylloides into a "borderline case" and subsequently a fibrosarcoma, makes it clear that fibrosarcoma of the breast cannot always be regarded as a tumor which has developed primarily from mesenchymal breast tissue. Just as a benign cystosarcoma can turn malignant if therapy is inadequate, it can also turn into a purely sarcomatous tumor. The clinical consequences of this are that a cystosarcoma should not only be shelled out, but excised generously, with a wide border of healthy tissue. The "borderline" type of cystosarcoma phylloides, or respectively the fibrosarcoma, requires far more aggressive therapy, i.e., mastectomy. In fibrosarcoma cases additional selective lymphadenectomy may be considered, although the metastasization pattern is predominantly hematogenic. Overall, however, the prognosis for fibrosarcoma cases is better than for those with breast cancer. PMID- 3026879 TI - Chronic effects of streptozotocin diabetes on myocardial sensitivity in the rat. AB - One month after streptozotocin treatment, basal rate in spontaneously beating right atria was decreased and basal developed force in electrically-driven right ventricular tissue was increased. Atrial sensitivity to the chronotropic effects of isoproterenol was not altered. In contrast, sensitivity in ventricular tissue to the inotropic effects of isoproterenol was decreased while sensitivity to calcium was increased. Associated with these changes was a decrease in myocardial beta-adrenoceptor density. Data obtained 3 and 6 months after streptozotocin treatment were similar to the observed alterations at 1 month. These results suggest that alterations in the chronotropic and inotropic responses that are expressed within 1 month after streptozotocin treatment do not significantly progress during the 6 months following induction of diabetes. They therefore reveal the independence of myocardial alterations from age of the animal and duration of diabetes (up to 6 months). PMID- 3026881 TI - Action of melanin-concentrating hormone (MCH) on teleost chromatophores. AB - The in vitro effects of synthetic salmon melanin-concentrating hormone (MCH) on chromatophores of four teleost species were studied. In the erythrophores of the platyfish (Xiphophorus maculatus) and the swordtail (Xiphophorus helleri), and in the xanthophores and amelanotic melanophores of the medaka (Oryzias latipes), pigment aggregation took place in response to MCH even in the absence of Ca2+. In contrast to this, the leucophores of the medaka responded to MCH by the pigment dispersion but only when Ca2+ was present. The motile iridophores of the blue damselfish (Chrysiptera cyanea), which play a predominant role in coloration and its changes, were not affected by the hormone. Pharmacological studies employing various blocking agents suggest that the pigment-aggregating action of MCH is probably mediated through specific receptors possessed by the erythrophores, xanthophores, or amelanotic melanophores, while the pigment-dispersing action on the leucophores might be revealed through the receptors for melanophore stimulating hormone (MSH). PMID- 3026882 TI - Effects of [D-Ala6, Pro9-NEt]-LHRH and catecholaminergic drugs on gonadotropin secretion and ovulation in the Chinese loach (Paramisgurnus dabryanus). AB - The effects of [D-Ala6,Pro9-NEt]-LHRH (LHRH-A) alone and in combination with drugs which influence the actions of dopamine or the synthesis of catecholamines on gonadotropin (GtH) secretion and ovulation in the loach (Paramisgurnus dabryanus) were investigated. LHRH-A alone stimulated an increase in serum GtH levels in the loach, but was a relatively ineffective treatment for the induction of ovulation. Injection of the dopamine receptor antagonist pimozide caused a marked potentiation of the GtH-release response to LHRH-A, and combined injections of pimozide and LHRH-A were an effective treatment for the induction of ovulation. Reserpine, a drug which causes depletion of catecholamines from presynaptic terminals, also caused a marked potentiation of the GtH-release response to LHRH-A and combined treatment induced ovulation. Similarly, administration of alpha-methyl-para-tyrosine to block conversion of tyrosine to L dopa, or carbidopa to block conversion of L-dopa to dopamine, potentiated the GtH release response to LHRH-A and induced ovulation. In contrast, the use of diethyldithiocarbamate, to block conversion of dopamine to norepinephrine, failed to augment the action of LHRH-A on GtH release and ovulation. The present results provide further evidence to suggest that dopamine functions as a gonadotropin release-inhibitory factor in teleosts, and demonstrate that the use of drugs which block either the synthesis or the actions of dopamine potentiates the action of LHRH-A in teleosts. PMID- 3026883 TI - On the stimulatory nature of the control of MSH secretion in ducks. AB - Though birds lack the pars intermedia of the hypophysis, their pituitary glands do secrete MSH. This hormone and ACTH are elaborated in special cells of the pars distalis called corticomelanotrops. The present study was designed to ascertain whether the release of MSH in aves is under either stimulatory or inhibitory hypothalamic control. Extracts of median eminence were injected in ducks and plasma MSH was observed to rise after the injections: on the other hand, when pituitaries were ectopically grafted, significant changes in the levels of circulating MSH were not detected. Twenty days after grafting, the transplants were extirpated and incubated in media containing median eminence extracts. The extracts stimulated the release of MSH not only from grafts but also from pieces of normotopic glands. The grafts showed cells which contained ACTH but not MSH; however, they contained small amounts of MSH, detectable by RIA. The administration of ergocryptine brought about the inhibition of MSH secretion in vivo, and it is suggested that this drug acted on hypothalamic structures rather than directly on the corticomelanotrops. On the basis of the preceding results, it is concluded for the first time that in ducks the release of MSH has stimulatory control from the hypothalamus, contrarily to that occurring in almost all the animals so far investigated. PMID- 3026884 TI - Effect of plasma lipoproteins in gonadotropin stimulation of 17 beta-estradiol production in the ovarian follicle of rainbow trout (Salmo gairdneri). AB - The effect of trout plasma lipoproteins on the production of 17 beta-estradiol by trout ovarian follicles is investigated in vitro. 17 beta-Estradiol secretion into the medium was assayed as a function of follicular diameter in the presence of lipoproteins with and without salmonid gonadotropin (SGA-GTH). The presence of very low-density lipoproteins (VLDL) + chylomicrons (Chy), low-density lipoproteins (LDL), and high-density lipoproteins (HDL) amplified the SGA-GTH effect at the lowest concentrations tested (less than 50 micrograms protein/ml). HDL is the most effective for increasing hormone accumulation on a microgram lipoprotein sterol basis. Autoradiography of 125I-labeled LDL showed that they were preferentially bound by thecal cells. Kinetics of 17 beta-estradiol release indicated that lipoprotein amplification occurred especially after 15 hr and subsequent metabolism of 17 beta-estradiol by follicular layers also led to an equilibrium. At the end of vitellogenesis apoprotein B lipoproteins (VLDL + Chy, LDL) apparently inhibited SGA-GTH stimulation. N',O'-Dibutyryl cAMP (10 mM) considerably stimulated 17 beta-estradiol production but lipoprotein amplification did not occur. Chloroquine (30 microM) inhibition of LDL and HDL amplification indicates that this process requires lysosomal degradation. Plasma lipoproteins in trout modulate SGA-GTH stimulation of 17 beta-estradiol production during exogenous vitellogenesis. Due to the ease and frequency with which the experiments can be carried out, the ovarian follicle of salmonids is an excellent model for the study of the role of lipoproteins in the regulation of ovarian steroids biosynthesis. PMID- 3026885 TI - Adrenocortical responses of pigeons (Columba livia) to treadwheel exercise. AB - Voluntary flight has previously been observed to stimulate adrenocortical activity in pigeons and in the present study increased circulating corticosterone levels were observed in adult feral pigeons forced to exercise in a treadwheel. The magnitude of the corticosterone response to treadwheel exercise was related to the speed of the moving drum, and hence to the work rate involved. Rapidly increasing the ambient temperature also stimulated corticosterone secretion in pigeons and masked the stimulatory effect of exercise. Treadwheel locomotion may serve as a laboratory model for the endocrine physiology of exercise and complements studies on flight. PMID- 3026886 TI - Plasma immunoreactive-growth hormone in domestic fowl: measurement by homologous and heterologous radioimmunoassays. AB - Concentrations of immunoreactive (IR) growth hormone (GH) in the plasma of domestic fowl have been measured by homologous and heterologous radioimmunoassays and the estimates compared. Both assays detected an age-related decline in the circulating IR-GH concentration, an increase in IR-GH secretion following TRH stimulation or fasting, and a fall in the IR-GH concentration following adrenocorticotropin administration. However, while the overall estimates of IR-GH concentration were significantly correlated, the magnitude of the changes in IR GH concentration determined by the homologous assay were far greater than those detected by the heterologous system, which failed to show any inhibitory effect of anesthesia or exogenous thyroid hormones on basal or stimulated IR-GH release. These results suggest that the heterologous GH radioimmunoassay lacks the sensitivity of the homologous chicken GH assay and that circulating GH in birds is probably composed of heterogeneous moieties with differing immunoreactivities with rat GH antibodies. PMID- 3026887 TI - Effects of air ions on the membrane Na, K-ATPase activity of L 1210 cells. AB - The effect of exposure of L 1210 mouse leukemia cells to artificially generated air ions on the activity of membrane Na, K-ATPase in the cells was investigated. The exposure of cells to air ions of both signs gave identical results, i.e. diminution of transport activity of the enzyme, measured by radioactive 86Rb transport into the cell (ouabain-dependent). Passive ouabain-independent transport of Rb+ into the cells remained unchanged in the air ion-treated cells, as did the passive efflux of the radioisotope from preloaded cells. The possible explanation of the phenomena observed is discussed. PMID- 3026888 TI - Heart sarcolemmal (Na+ + K+)-ATPase has an essential amino group in the potassium binding site on the enzyme molecule. AB - Selective modification of primary amino groups of (Na+ + K+)-ATPase by trinitrobenzene sulfonic acid (TNBS) resulted in a considerable inhibition of the specific activity of the enzyme. Investigation by means of enzyme and sorption kinetics of activation of heart sarcolemmal (Na+ + K+)-ATPase by its monovalent cationic ligands added simultaneously with TNBS revealed: a considerable competition between K+-ions and TNBS for the potassium binding site on the enzyme molecule; a non-competitive type of inhibition of Na+-induced activation of the enzyme. Both, potassium and sodium ions depressed, and magnesium ions enhanced the initial rate of TNBS-sorption; however, none of the above cations influenced the equilibrium value of TNBS sorption onto isolated sarcolemmal membranes. Ouabain, on the other hand, did not inhibit the initial rate and decreased the equilibrium value of TNBS sorption onto the membranes. The results obtained enabled the identification of an essential amino group in the potassium binding site of the (Na+ + K+)-ATPase molecule. PMID- 3026889 TI - Effect of calcium on the structure-function relationship of (Na+ + K+)-ATPase in cardiac sarcolemma. AB - Calcium-induced changes in (Na+ + K+)-ATPase activity and structural changes of membrane bound proteins in rat heart sarcolemma were investigated. Increasing concentrations of Ca2+ (0.1-8.0 mmol.l-1) gradually inhibited the (Na+ + K+) ATPase activity and decreased the alpha-helix content of sarcolemmal proteins. Mathematical and graphical analysis of observed data yielded a quantitative relationship between Ca2+-induced changes in (Na+ + K+)-ATPase activity and the secondary structure of membrane proteins in cardiac sarcolemma. PMID- 3026890 TI - Effect of temperature on Ca-ATPase from sarcoplasmic reticulum membranes: ESR studies. AB - Using spin-labeled fatty acid derivatives and maleimide, the effect of temperature on the structural state of various parts of the lipid bilayer of sarcoplasmic reticulum (SR) membranes and the segmental motion of the Ca-ATPase molecule were investigated. The mobility of the spin probes localized in the hydrophobic zone and the outer part of the SR membrane was shown to increase with a rise in temperature from 4 to 44 degrees C, the temperature of 20 degrees C being critical for these changes. In the presence of ATP, critical changes in the spin probe mobility occur at lower temperatures, while in the presence of ATP and Ca2+ they are observed at 20 degrees C for a spin probe localized in the outer part of the SR membrane. The mobility of a spin probe localized in the hydrophobic part of the membrane increases linearly with a rise in temperature. In the absence of ligands, the segmental motion of Ca-ATPase changes linearly within a temperature range of 10-30 degrees C. However, when ATP alone or ATP and Ca2+ are simultaneously added to the incubation mixture, the protein mobility undergoes critical changes at 20 degrees C. The Arrhenius plots for ATPase activity and Ca2+ uptake rate in SR membrane preparations also have a break at 20 degrees C. It is assumed that changes in the structural state of membrane lipids produce conformational changes in the Ca-ATPase molecule; the enzyme seems to be unsensitive to the structural state of the membrane lipid matrix in the absence of the ligands. PMID- 3026891 TI - Sequence and transcripts of the bacteriophage T4 DNA repair gene uvsY. AB - We have cloned, sequenced and analyzed transcription of the phage T4 uvsY gene. This gene is transcribed from a single gp MotA-dependent middle promoter to give a major transcript of approximately 930 nucleotides and a minor transcript of approximately 620 nucleotides. All in vivo and in vitro uvsY transcripts show anomalous migration in agarose gels. The uvsY transcript contains an open reading frame coding for an 137 amino acid [15.8 kilodaltons (kD)] UvsY protein and two unidentified open reading frames, ORF UvsY.-1 (9.0 kD) and ORF UvsY.-2 (6.0 kD). Our DNA sequence differs in only three places from that published by TAKAHASHI et al. However, one of these changes alters the predicted carboxy terminus of the UvsY protein. Marker rescue experiments map gene 25 to the region upstream of uvsY. Gene 25 is likely, although not certain, to correspond to an ORF that is found upstream from uvsY and is translated in the same direction. PMID- 3026892 TI - Coincident gene conversion events in yeast that involve a large insertion. AB - In yeast, spontaneous gene conversion events involving sites that are far apart (16 cM) occur 1000 times more frequently in mitotic cells than is expected for two independent acts of recombination. It has been proposed that a major portion of these could be due to a long, continuous heteroduplex intermediate. We have examined this possibility in further detail by introducing, via transformation, a large plasmid insertion between the LEU1 and TRP5 loci and studying its behavior among coincident convertants involving the flanking sites. Among such convertants, there is frequent loss of the plasmid when it is present in hemizygous or homozygous configuration. Our results could support the long heteroduplex model for coincident recombination events, but only if novel assumptions regarding the formation and fate of mismatched DNA are made. Therefore, an alternative model that proposes multiple, concerted recombination events is discussed. PMID- 3026894 TI - [Bacterial transposons and the mechanisms of their transposition]. AB - The published data on molecular mechanisms of transposons movement inside and between genomes are reviewed. The replicative mechanism of transposition of the family of Tn3-like elements is discussed, as well as the modes of bacteriophage Mu, Tn9, Tn10, Tn903 transposition. The factors affecting the choice of transposition pathways are analysed. PMID- 3026895 TI - [2 types of molecular structure (composition) of a genome in one species of transposable bacteriophages of Pseudomonas aeruginosa]. AB - Comparison of heteroduplexes (HD) between DNAs of different transposable phages of Pseudomonas aeruginosa belonging to two previously described subgroups (D3112 and B3) revealed two types of structure (composition) of the bacteriophages, designated "type A" and "type B". The properties of genome structure of type A (phages of D3112 subgroup) are as follows: high level of conservation (up to 70% of genomes of different phages are represented as blocks of homologous DNA sequences); substitutions in genomes revealed as nonhomology regions in HD are, as a rule, small and located in certain sites; the distribution of the nonhomologous regions in HD of these phages is highly reproducible in independent experiments. Bacteriophages of subgroup B3 have genomes of type B: only a small part (approx. 30%) of genomes retain homology general for all of the phages; the nonhomologous regions are distributed in a large number of sites in HD; the sizes of nonhomologous regions are substantially larger than for the phages of subgroup D3112; distribution of the regions in HD is highly variable, which is characteristic of DNAs with partial homology. There is no difference between genomes of types A and B in G + C content (approx. 61-63%). Viable recombinants can be formed in crosses between phages of different genome types not only in regions with earlier revealed large DNA/DNA homology (right ends of genomes), but also in central portions of the genomes. Nevertheless, functional incompatibility of some regions of phage genomes of types A and B was demonstrated. PMID- 3026893 TI - Molecular population genetics of the alcohol dehydrogenase gene region of Drosophila melanogaster. AB - Variation in the DNA restriction map of a 13-kb region of chromosome II including the alcohol dehydrogenase structural gene (Adh) was examined in Drosophila melanogaster from natural populations. Detailed analysis of 48 D. melanogaster lines representing four eastern United States populations revealed extensive DNA sequence variation due to base substitutions, insertions and deletions. Cloning of this region from several lines allowed characterization of length variation as due to unique sequence insertions or deletions [nine sizes; 21-200 base pairs (bp)] or transposable element insertions (several sizes, 340 bp to 10.2 kb, representing four different elements). Despite this extensive variation in sequences flanking the Adh gene, only one length polymorphism is clearly associated with altered Adh expression (a copia element approximately 250 bp 5' to the distal transcript start site). Nonetheless, the frequency spectra of transposable elements within and between Drosophila species suggests they are slightly deleterious. Strong nonrandom associations are observed among Adh region sequence variants, ADH allozyme (Fast vs. Slow), ADH enzyme activity and the chromosome inversion ln(2L)t. Phylogenetic analysis of restriction map haplotypes suggest that the major twofold component of ADH activity variation (high vs. low, typical of Fast and Slow allozymes, respectively) is due to sequence variation tightly linked to and possibly distinct from that underlying the allozyme difference. The patterns of nucleotide and haplotype variation for Fast and Slow allozyme lines are consistent with the recent increase in frequency and spread of the Fast haplotype associated with high ADH activity. These data emphasize the important role of evolutionary history and strong nonrandom associations among tightly linked sequence variation as determinants of the patterns of variation observed in natural populations. PMID- 3026896 TI - [Characteristics of derivatives of the plasmid RP4 in a broad range of hosts with altered properties of maintenance and inheritance]. AB - Hydroxylamine-induced mutants of the plasmid pPD6 (8.4 kb) were isolated which are resistant to high doses of tetracycline. One of the plasmids studied--pPD21 is a multicopy mutant, another one, pPD12 is a dimeric form of the pPD6 plasmid. The pPD12 plasmid is very unstable, its derivative, pPD13 spontaneous mutant acquiring stability but not the ability to resolve DNA multimeric forms into monomeric forms. Multicopy bireplicon pPD619 plasmid was constructed by joining in vitro pPD6 and pUC19 plasmids. Removing the replicon pUC19 from the bireplicon plasmid gives a new low-copy plasmid pPD620. All of the plasmids constructed were mobilized by the conjugative pRK2013 plasmid into the strains of Escherichia coli, Pseudomonas aeruginosa and Agrobacterium tumefaciens. The pPD6 plasmid and its derivatives can be used as cloning vectors. PMID- 3026897 TI - [Nah-genes of Pseudomonas putida: molecular genetic analysis of the plasmid pBS286]. AB - The hybridization and restriction analysis of the plasmid pBS286 (73 Kb, the P-9 Inc group) as well as parental plasmids NPL-1, NPL-41 demonstrated that pBS286 plasmid (delta NPL-41::TnA) with the constitutive synthesis of naphthalene dioxygenase carried genes for naphthalene oxidation to salicylate and those participating in degradation of catechol. Restriction map of pBS286 using XhoI restriction endonuclease and that of the nah region using EcoRI, BamHI, SalI and XhoI were established. Structural peculiarities of nah genes from pBS286 are compared with previously described NAH7. Some nah genes were localized. An inverted DNA segment involved in nah gene regulation was shown to be closely linked to a proximal part of the nah1 operon or overlapped. Possible occurrence of a regulatory R locus in this region is suggested. PMID- 3026898 TI - [Cloning of genes for proline biosynthesis in Escherichia coli]. AB - Restriction map of Escherichia coli chromosome fragment (7.4 MD) carrying proAB genes was constructed. Localization of proA and proB genes on the cloned chromosome fragment was determined by complementation test and the measuring of glutamylkinase activity (proB gene product). ProA and proB genes were cloned separately on multicopy plasmids of alternative orientation and their expression being, probably, under the control of their own regulatory regions, studied. PMID- 3026899 TI - [Cloning and characteristics of recA gene in Pseudomonas aeruginosa]. AB - A recA-like gene from Pseudomonas aeruginosa was cloned and identified by means of interspecific complementation of gene recA repair defect in Escherichia coli. The gene was mapped in the PvuII-HindIII Ps. aeruginosa chromosome fragment of 1.5 kbp in length. Having been recloned in pUC18 or 19 plasmids in either of possible orientations, this fragment was shown to complement three different defects of E. coli recA mutants: in repair, recombination and SOS functions. PMID- 3026900 TI - [RecE-independent recombination of plasmids in Bacillus subtilis cells]. AB - The pUB110 and pE194 plasmid cointegrates have been isolated and examined in rec+ and recE4 strains of Bacillus subtilis. Cointegrates were shown to be formed by recombination at the specific site present on both parental plasmids as a short region of homology designated RSA. The RSA consists of 63 nucleotides in pE194 and 49 in pUB110; the length of its fully conserved core segment is 10 nucleotides. All cointegrates examined were formed by single crossover event taking place within the core segment, and as a result they have identical nucleotide sequences of recombination junctions. No conversion of mismatched base pairs to nucleotide sequences originally belonging to one of the parental plasmids was found. Though the action of RecE gene did not affect the frequency of cointegrate formation, it was reduced in rec149 host by one order of magnitude. Cointegrates retained their stability during transformation. PMID- 3026901 TI - Secretion of enzymatically active human renin from mammalian cells using an avian retroviral vector. AB - Recombinant plasmid-based retroviral expression vectors were constructed using a modified spleen necrosis virus (SNV) containing the Herpes simplex virus thymidine kinase gene promoter controlling the expression of the Tn5 neomycin phosphotransferase II gene (NPTII gene). The human renin (HRn) gene (hrn) was inserted into the 5' end of the SNV sequences such that in concatemeric plasmid DNA its expression was controlled by the strong promoter in the SNV long terminal repeat (LTR). Dog cells transfected with the concatemeric plasmid DNA secreted a small amount of a HRn-like 43-kDa protein. After cotransfection of chicken cells with concatemeric plasmid DNA and proviral DNA of reticuloendotheliosis virus strain A, infectious stocks of viruses were recovered. Cells infected with the virus carrying the viral LTR-hrn gene oriented for expression secreted the 43-kDa HRn-like protein at about 100-fold higher levels than the cells transfected with the plasmid DNAs. Biological activity of secreted HRn was determined by measuring levels of angiotensin I generated by incubating culture media with either a porcine or human angiotensinogen substrate. Infected dog cells produce about 40 ng of enzymatically active HRn per 10(6) cells per 24 h. These data indicate that retroviral expression vectors provide a good system for obtaining the secretion of high levels of enzymatically active heterologous proteins from mammalian cells. PMID- 3026903 TI - An efficient chloramphenicol-resistance marker for Saccharomyces cerevisiae and Escherichia coli. AB - Chloramphenicol (Cm) was demonstrated to be a suitable selective agent for the plasmid-mediated transformation of haploid and polyploid strains of Saccharomyces cerevisiae. A yeast/Escherichia coli shuttle Cm-resistance (CmR) marker was constructed by inserting the CAT coding sequence from Tn9, and its associated bacterial ribosome-binding site, between a modified yeast ADC1 promoter and CYC1 terminator. When present on a 2 microns-based replicating plasmid, this marker transformed yeast as efficiently as the auxotrophic markers TRP1 and LEU2. When included in an integrating vector, single-copy transformants were formed as efficiently as with LEU2 and HIS3. Industrial yeast strains were transformed with both the replicating and integrating plasmids. The CmR marker could also efficiently transform E. coli. This versatile and efficient performance is currently unique for a yeast dominant marker. PMID- 3026902 TI - Cloning a cDNA for Drosophila melanogaster urate oxidase. AB - A cDNA library from third-instar larval Malpighian tubules of Drosophila melanogaster was constructed and screened for urate oxidase (UO) clones by hybridization selection. The coding sequence for UO was mapped by in situ hybridization to position 28C on the left arm of chromosome 2. The UO activity in Drosophila shows a complex developmental profile. A UO cDNA was used as a probe of Northern blots of poly(A) + RNA from various stages of development. The data show that there is a direct correlation between the transcriptional activity of the UO locus as evidenced by the quantitative changes of UO mRNA and the levels of UO activity and protein during development. PMID- 3026907 TI - A plasmid expression vector that permits stabilization of both mRNAs and proteins encoded by the cloned genes. AB - Two new expression vectors have been constructed to take advantage of several useful properties of bacteriophage T4-infected Escherichia coli. These plasmids, pRDB8 and pRDB9, contain the promoter region and start codon of T4 gene 32, a contiguous multiple cloning site (MCS), and translation and transcription termination signals. DNA fragments inserted into the MCS are transcribed and translated at a high level in both uninfected and phage T4-infected cells. Furthermore, the extreme stability of the hybrid mRNA after infection permits the specific biosynthetic labeling of the protein encoded by the cloned gene. In addition, the cloned gene product is stabilized, since the host-mediated degradation of foreign proteins is inhibited by phage infection. The properties of this expression system were demonstrated with the constant region of a rabbit immunoglobulin lambda light chain (C lambda) gene. Although proteolytic degradation of the C lambda fusion protein was rapid in uninfected cells, degradation was blocked in phage-infected cells and the protein accumulated in greater amounts. PMID- 3026904 TI - Transient expression of murine interferon-alpha genes in mouse and monkey cells. AB - The coding regions of murine interferon-alpha (IFN-alpha) genes were combined with promoter and 3'-noncoding sequences from other eukaryotic genes. Transient expression of these fusion genes was achieved in monkey COS cells and in a mouse cell line (TOP cells) expressing polyoma virus (Py) large T antigen constitutively. The efficiency of the different expression plasmids was determined by measuring the amount of IFN secreted into the medium. Replacement of the 3'-noncoding region of an IFN-alpha gene by that of the rabbit beta-globin gene resulted in a fourfold higher IFN-alpha production. The SV40 early promoter and the Moloney murine leukemia virus (MoMLV) long terminal repeat (LTR) produced similar amounts of IFN-alpha in COS cells. However, a tandem combination of the SV40 enhancer/early promoter and the mouse metallothionein-I promoter appeared fivefold more active than the SV40 early promoter. In TOP cells the MoMLV LTR was found to be threefold more active than the Py early promoter. PMID- 3026908 TI - Luciferase genes cloned from the unculturable luminous bacteroid symbiont of the Caribbean flashlight fish, Kryptophanaron alfredi. AB - Light organs of anomalopid (flashlight) fish contain luminous bacteroids that have never been cultured and, consequently, have been difficult to study. We have characterized the luciferase (lux) region of DNA extracted from light organs of the Caribbean flashlight fish Kryptophanaron alfredi by hybridization of cloned Vibrio harveyi lux genes to restriction-endonuclease-digested, light organ DNA. Comparison of the hybridization pattern of light organ DNA with that of DNA of a putative symbiotic isolate provides a method for identifying the authentic luminous symbiont regardless of its luminescence, and was used to reject one such isolate. Light organ DNA was further used to construct a cosmid clone bank and the luciferase genes were isolated. Unlike other bacterial luciferase genes, the genes were not expressed in Escherichia coli. When placed under the control of the E. coli trp promoter, the genes were transcribed but no luciferase was detected, suggesting a posttranscriptional block to expression. PMID- 3026906 TI - Sequencing and heterologous expression of the gene encoding penicillin V amidase from Bacillus sphaericus. AB - The Bacillus sphaericus gene encoding penicillin V amidase, which catalyzes the hydrolysis of penicillin V, has been characterized. The entire nucleotide sequence of the coding region, as well as 5'- and 3'-flanking regions, was determined using an improved sequencing strategy. The deduced amino acid sequence suggests a protein consisting of 338 residues with an Mr of 37,500. The ATG initiator codon is preceded by a putative ribosome-binding site, typical for genes of Gram-positive origin. High expression of the gene was obtained in Escherichia coli using an inducible promoter, showing that the gene product is stable in this heterologous host. PMID- 3026909 TI - Expression of anti-sense mRNA in H-ras transfected NIH/3T3 cells does not suppress the transformed phenotype. AB - We have attempted to reverse the transformed phenotype of cells expressing the H ras oncogene. A plasmid in which the first exon of the H-ras oncogene was coupled to the SV40 early promoter in an anti-sense orientation was constructed. This construct was introduced into a clone of H-ras-transformed NIH/3T3 cells. Simultaneous expression of both the SV40 anti-sense construct and H-ras was observed. Anti-sense RNA was present in a 10-20-fold excess over sense H-ras RNA. Only a small fraction of the cytoplasmic RNA was present in a sense: anti-sense duplexed form. The expression of anti-sense H-ras RNA was not accompanied by a phenotypic reversion of transformed cells. The only phenotypic reversion we observed was accompanied by a loss of transfected H-ras sequences. The loss of transfected H-ras sequences occurs with a high frequency in cells supertransfected with the SV40 anti-sense construct. PMID- 3026905 TI - Cloning and expression in biologically active form of the gene for human interferon alpha 2 in Streptomyces lividans. AB - A fragment of human DNA encoding the mature form of interferon alpha 2 (hIFN alpha 2), and carrying both an in-phase ATG initiation codon and the ribosome binding site (RBS) of the Escherichia coli membrane lipoprotein gene (lpp), was fused to the aminoglycoside phosphotransferase gene (aph) promoter (aphP) from Streptomyces fradiae. When this construction was inserted, in the two possible orientations, in the Streptomyces plasmid pIJ702, plasmids pNIS19 and pNIS91 were obtained. A 20-kDa polypeptide that immunoreacted with an hIFN-alpha 2 monoclonal antibody was expressed in S. lividans clones carrying these plasmids. Moreover, these clones contained an intracellular antiviral activity similar to that of hIFN-alpha 2. When plasmids pNIS19 and pNIS91 were deprived of the aphP no expression of activity was found. Therefore, it is concluded that the hIFN gene can be efficiently expressed in Streptomyces as directed by the aph gene promoter. PMID- 3026910 TI - Insertion element IS1 can generate a 10-base pair target duplication. AB - Transposable element IS1 is known to generate mainly 9-bp and occasionally 8-bp target duplications upon transposition. We have isolated a plasmid pBR322 derivative having IS1 inserted into a site between the promoter and the structural gene for tetracycline resistance. DNA sequence analysis revealed that integration of this IS1 resulted in a 10-bp target duplication. PMID- 3026911 TI - A simple and rapid nucleotide sequencing strategy and its application in analyzing a rice histone 3 gene. AB - An improved rapid method for sequencing a target DNA is described. A new plasmid, pAA-PZ1, which contains the origin of replication from phage M13 and a portion of the Tn9 transposon was constructed. A long fragment of target DNA cloned into this vector is progressively shortened in vivo from one end by transposon mediated deletions. The plasmids carrying different lengths of target DNA are then made into single-stranded DNA in the same host upon infection with an M13 phage and their sequence is determined using the dideoxynucleotide chain termination method. This method bypasses the in vitro enzymatic manipulations for progressive deletions and requires no subcloning. Using this strategy, we sequenced 1.3 kb of rice DNA containing a histone 3 gene within three weeks. PMID- 3026912 TI - Structure of the two genes coding for polypeptide chain elongation factor 1 alpha (EF-1 alpha) from Saccharomyces cerevisiae. AB - Polypeptide chain elongation factor 1 alpha (EF-1 alpha) of Saccharomyces cerevisiae is encoded by two distinct genes designated EF1 alpha A and EF1 alpha B [Nagata et al., EMBO J. 3 (1984) 1825-1830]. Both genes were cloned, and their nucleotide (nt) sequences were determined [see also Schirmaier and Philippsen, EMBO J. 3 (1984) 3311-3315, and Cottrelle et al., J. Biol. Chem. 260 (1985) 3090 3096]. They contain an open reading frame of 1374 nt coding for an identical protein of 458 amino acid residues, although their nt sequences differed at two positions. In this paper, we determined their 5'- and 3'-flanking sequences which were considerably different each other. From the S1 nuclease mapping of mRNA, both genes are found to be expressed almost to the same extent in exponentially growing cells. The transcription start points for EF1 alpha A and EF1 alpha B mRNAs were precisely located by primer extension procedure at 32 and 23 nt upstream of the start codons, respectively. The sequence which commonly exists in the 5'-flanking regions of ribosomal protein genes of S. cerevisiae was also present in the two EF1 alpha genes. PMID- 3026913 TI - Deletion loop mutagenesis of the nifL promoter from Klebsiella pneumoniae: role of the -26 to -12 region in promoter function. AB - Nine single C-to-T transitions were introduced into the -26 to -12 region of the Klebsiella pneumoniae nifL promoter by bisulphite mutagenesis of M13 heteroduplexes containing a 15 nucleotide single-stranded loop. Mutant promoter fragments were inserted into translational lac fusion vectors to utilise beta galactosidase activity as a measure of promoter efficiency. Mutations in invariant nucleotides found in the consensus sequence for nif promoters gave a strong 'down' promoter phenotype with respect to transcriptional activation. Mutations in semi-conserved residues had a much weaker down phenotype, whereas a mutation which increased homology to the consensus sequence enhanced promoter strength. One mutant showed increased activation by ntrC and decreased activation by nifA. PMID- 3026914 TI - The mosaic cox1 gene in the mitochondrial genome of Schizosaccharomyces pombe: minimal structural requirements and evolution of group I introns. AB - The gene encoding subunit 1 of cytochrome oxidase (cox1) in the fission yeast Schizosaccharomyces pombe is polymorphic. In strain 50 it contains two group I introns with open reading frames (ORFs) in phase with the upstream exons (Lang, 1984). In strain EF1 two additional very short group I introns which do not possess ORFs were detected by DNA sequencing. These two introns (AI2a and AI3) share distinct characteristics concerning their nucleotide sequence and secondary structure and are located at identical positions as the introns AI4 and AI5 beta, respectively, in the cox1 gene of Saccharomyces cerevisiae. The sequence homology of the cob and cox1 genes around the splice points of introns AI2a, AI4, and BI4 (cob intron 4) might reflect horizontal gene transfer between the distantly related species S. pombe and S. cerevisiae. PMID- 3026915 TI - Yeast shuttle and integrative vectors with multiple cloning sites suitable for construction of lacZ fusions. AB - We report yeast/Escherichia coli shuttle vectors suitable for fusing yeast promoter and coding sequences to the lacZ gene of E. coli. The vectors contain a region of multiple unique restriction sites including EcoRI, KpnI, SmaI, BamHI, XbaI, SalI, PstI, SphI and HindIII. The region with the unique cloning sites has been introduced in both orientations with respect to lacZ and occurs proximal to the eighth codon of the gene. All the restriction sites have been phased to three different reading frames. Two series of vectors have been constructed. The first series (YEp) has two origins of replication (ori), i.e., of the yeast 2 mu circle and of the ColE1 plasmid of E. coli, and can therefore replicate autonomously in both organisms. These shuttle vectors also have the ApR gene of E. coli and either the yeast LEU2 or URA3 genes to allow for selection of both E. coli and yeast transformants. The second series of vectors (YIp) are identical in all respects to the YEp vectors except that they lack the 2 mu ori. The YIp vectors can be used to integrate lacZ fusions into yeast chromosomal DNA. None of the vectors express beta-galactosidase (beta Gal) in yeast or E. coli in the absence of inserted yeast promoter sequences. The 5'-nontranslated sequences and parts of the coding sequences of various yeast genes have been cloned into representative lacZ fusion vectors. In-frame gene fusions can be detected by beta Gal activity when either yeast or E. coli clones are plated on media containing XGal indicator. Quantitative determinations of promoter activity were made by colorimetric assay of beta Gal activity in whole cells. Fusion of the yeast CYC1 gene to lacZ in one of the vectors allowed detection of regulated expression of this gene when cells were grown under conditions of catabolite repression or derepression. PMID- 3026916 TI - Direct expression of urogastrone gene in Escherichia coli. AB - Human epidermal growth factor (urogastrone; UG) is a 53-amino acid polypeptide hormone. A 192-bp DNA fragment containing the coding sequence for methionyl UG (Met-UG) and the ribosome-binding site (RBS) was chemically synthesized and placed downstream from the promotor for the Escherichia coli outer-membrane lipoprotein gene (lpp) on a plasmid. E. coli cells harboring the plasmid directed the synthesis of Met-UG at 10(2)-10(3) molecules per cell. Next, the coding sequence for Met-UG was inserted in a runaway-replication plasmid and expressed under the control of the lpp promoter and the RBS derived from bacteriophage Mu cII gene. Upon heat induction, the cells harboring the recombinant plasmid synthesized 10(5) molecules of Met-UG per cell. PMID- 3026917 TI - Synthesis of fusion and mature murine alpha interferons in Escherichia coli. AB - Four murine interferon-alpha (MuIFN-alpha) genes (alpha 1, alpha 4, alpha 5, alpha 6) were previously identified and characterized. The coding regions of these IFN-alpha genes were inserted into bacterial expression vectors behind the lpp promoter under the control of the lac promoter-operator region, resulting in fusion peptides containing additional N-terminal amino acids (aa). Plasmids coding for the expression of mature IFN-alpha 1 and alpha 5 were also constructed using the same vector system, by inserting a 30-bp synthetic oligodeoxynucleotide, which contains a stop codon for the lpp gene, a ribosome binding sequence and an ATG start codon for the IFN peptides. The amounts of IFN polypeptides synthesized in Escherichia coli were estimated in the maxi-cell system and their biological activities were measured on mouse and other mammalian cells. The yields of mature IFN produced in this vector were 2 to 4 X 10(6) units/liter; the antiviral activity of the majority of the MuIFNs on human and bovine cells was 100- to 1000-fold lower than on mouse cells. IFN-alpha 4, which contains an internal deletion of 5 aa, showed a lower antiviral activity than other MuIFNs on mouse cells. PMID- 3026918 TI - Integration of the delta-endotoxin gene of Bacillus thuringiensis into the chromosome of root-colonizing strains of pseudomonads using Tn5. AB - The delta-endotoxin gene (tox) from Bacillus thuringiensis subsp. kurstaki HD-1 was cloned into Tn5 and the resulting Tn5-tox element transposed from a vector plasmid into the chromosome of six corn-root-colonizing strains of Pseudomonas fluorescens and Agrobacterium radiobacter. Chromosomal integration of the tox gene maximized stability and minimized the potential for horizontal transfer of the tox gene to other bacterial species. Expression of the tox gene was demonstrated by Western blot analysis and by toxicity against larvae of the tobacco hornworm (Manduca sexta). The method described illustrates how a given gene can be stably integrated into the chromosome of diverse bacterial species. PMID- 3026919 TI - An improved filamentous helper phage for generating single-stranded plasmid DNA. AB - Gene cloning in plasmid vectors that contain a filamentous phage intergenic region presents several advantages. However, technical difficulties have been a problem, primarily low yields of packaged single stranded (ss) plasmid DNA from the rapid, small scale procedures usually employed, and ambiguities in sequencing reactions attributed to the contamination by helper phage ss DNA. We report here the construction and some properties of a new f1 helper phage. Using this phage, R408, plasmid ss DNA is packaged and exported preferentially over phage ss DNA, and the absolute yield of plasmid ss DNA is usually increased. PMID- 3026920 TI - Reconstitution of an operon from overlapping fragments: use of the lambda SV2 integrative cloning system. AB - We have used the lambda SV2 system [Howard and Gottesman. In Gluzman (Ed.), Eukaryotic Viral Vectors. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, 1982, pp. 211-216; in Inouye, M. (Ed.) Experimental Manipulations of Gene Expression. Academic Press, New York, 1983, pp. 137-153] to reconstitute the Salmonella typhimurium his operon from overlapping fragments. lambda SV2 can be propagated as an autonomously replicating plasmid or as a prophage integrated in the Escherichia coli chromosome at the lambda attachment site; our reconstitution was accomplished in the integrated state. We first inserted a portion of the his operon into lambda SV2 and integrated the resulting plasmid by site-specific recombination into the E. coli chromosome. This was achieved by brief induction of a resident prophage. The lysogen was then transformed with DNA from a lambda SV2 clone carrying the remainder of the his operon on an overlapping DNA fragment. The second plasmid was forced to integrate into the first by homologous recombination. When this recombination occurs at the his overlap, a lysogen carrying two lambda SV2 prophages is produced. One prophage carries the entire his operon and the other carries the his overlap region. The latter is removed by site-specific recombination, permitting further contiguous sequences to be sequentially added to the remaining prophage. This method should be applicable for the reconstitution and maintenance of large genes or gene clusters in the E. coli genome. PMID- 3026922 TI - Mutations resulting in promoter-like sequences which enhance the expression of araC in Salmonella typhimurium. AB - The araC gene in Salmonella typhimurium is autogeneously regulated. Nine non-self regulated mutants were isolated by selecting for increased expression of an araC lacZ fusion in the presence of a repressing AraC protein. S1 mapping experiments demonstrated that the effect of the mutations was to increase the amount of araC mRNA in the cell. The 5'-end of the major araC transcript was the same in the wild-type (wt) and mutant strains. DNA sequence analysis showed that all nine mutations occurred in the araC promoter. Two mutations were a G-to-T transversion at position -25, six were a G-to-T transversion at position -47 and one was a single bp deletion at position -83. The data suggest that the mutations have created a new RNA polymerase binding site which enhances transcription from the wt start point. PMID- 3026923 TI - Plasmid map: a microcomputer program for display and storage of plasmid data. AB - We describe a plasmid map program which runs on an IBM PC microcomputer and facilitates the drawing of circular plasmid maps. The user enters information from the keyboard in the form of restriction enzyme sites, genes and their locations, and other plasmid markers such as promoters, origins, or other sites. This information can then be stored in a file for future reference. The plasmid map can be displayed on the screen, printed on a dot-matrix printer, or plotted on a Hewlett Packard HP7475A plotter. PMID- 3026921 TI - Use of a modified Escherichia coli trpR gene to obtain tight regulation of high copy-number expression vectors. AB - It has been found that with high-copy-number vectors utilising the Escherichia coli trp promoter the amount of repressor protein produced from the single chromosomally located trpR gene is inadequate for tight repression to be obtained. An attempt has been made to overcome this problem by inserting the trpR gene in cis into the expression vector. This proved unsuccessful because transcription from the trp promoter of such a plasmid could not be induced with 3,beta-indole acrylic acid, probably because the trpR gene is autogenously regulated. However, it was found that when the natural trpR promoter was replaced with a relatively weak constitutive promoter a useful self-repressible vector could be formed. A modified trpR gene of this type has been used to obtain tightly controlled expression of human interferon-beta (IFN-beta) from a vector having a copy number of 400. Tight regulation is particularly important in this case as IFN-beta is highly toxic to the E. coli cell. PMID- 3026924 TI - Nucleotide sequence of the Erwinia chrysanthemi NCPPB 1066 L-asparaginase gene. AB - The complete nucleotide sequence of the Erwinia chrysanthemi NCPPB 1066 gene coding for the chemotherapeutic enzyme L-asparaginase has been determined. The structural gene consists of an open reading frame commencing with an ATG start codon of 1044 bp followed by a TGA stop codon. Confirmation of the nucleotide sequence was obtained by comparing the predicted amino acid (aa) sequence with that derived by N-terminal aa sequencing of the purified protein. The gene has been shown to code for a 21-aa signal peptide at its N terminus which closely resembles the signal peptides of other secreted proteins. In common with highly expressed Escherichia coli genes, little use is made of modulator codons. The predicted aa sequence of the enzyme exhibits 46% identity with the determined primary sequence of the E. coli L-asparaginase, although the predicted secondary structure of both proteins indicates more extensive homology. Downstream of the TGA stop codon is a G + C-rich region of dyad symmetry (delta G = -25.4 kcal) characteristic of E. coli Rho-independent transcription terminators. Upstream of the structural gene there are no sequences which bear a strong resemblance to the consensus -35 and -10 regions of E. coli promoters. A sequence is present (CTGGCTCTCCTCTTGAT), however, which exhibits strong homology to the nif promoter consensus sequence (CTGGCACN5TTGCA). Upstream of this region is a sequence which strongly resembles the consensus sequence for promoter regions which are subject to catabolite repression. PMID- 3026925 TI - Cloning and complete nucleotide sequence of the Bacillus stearothermophilus tryptophanyl tRNA synthetase gene. AB - The Bacillus stearothermophilus NCA 1503 tryptophanyl tRNA synthetase (WTS; EC 6.1.1.2) gene has been cloned in Escherichia coli and the amino acid (aa) sequence of the enzyme deduced unequivocally from the DNA sequence of the cloned gene. The predicted aa sequence of the WTS enzyme agrees with the previously determined aa sequence except that the DNA sequence indicates a third Arg residue at the C terminus of the enzyme over the two Arg residues indicated by sequencing the protein itself. The trpS gene consists of a 984-bp open reading frame commencing with an ATG start codon and ending with a TAA stop codon. Putative transcriptional promoters, a Shine-Dalgarno sequence and a transcription terminator have been identified. Thus the trpS gene probably constitutes a single transcriptional unit. PMID- 3026926 TI - Cloning, expression and complete nucleotide sequence of the Bacillus stearothermophilus L-lactate dehydrogenase gene. AB - The structural gene for L-lactate dehydrogenase (LDH; EC 1.1.1.27) from Bacillus stearothermophilus NCA 1503 has been cloned in Escherichia coli and its complete nucleotide sequence determined. The predicted amino acid (aa) sequence of the LDH enzyme agrees with the previously determined aa sequence except to three positions: aa 125 and 126, Ser-Glu, are inverted whilst His at position 130 has been replaced by Ser in our sequence. The lct gene consists of an open reading frame (ORF) commencing from the ATG start codon of 951 bp followed by a TGA stop codon. Upstream from the start codon is a strong (delta G = -14.4 kcal) Shine Dalgarno (SD) sequence, a feature typical of Gram-positive ribosome binding sites. Putative RNA polymerase recognition signals (-35 and -10 regions) have been identified upstream from the lct structural gene but there are no structures resembling Rho-independent transcription termination signals downstream from the TGA stop codon. Two further ORFs, preceded by SD sequences, are present downstream from the lct gene. Thus the lct gene may constitute the first gene of an operon. Subclones of the lct gene have been constructed in the expression plasmid pKK223-3 and the LDH enzyme produced in soluble form at levels of up to 36% of the E. coli soluble cell protein. PMID- 3026927 TI - Plasmids allowing transcription of cloned DNA by Salmonella typhimurium phage SP6 RNA polymerase to produce RNAs with authentic 5'-terminal sequences. AB - We wished to determine whether there is any specific sequence downstream of the start point of the SP6 promoter which is required for its function in the plasmid pSP64 (Melton et al., 1984). Lack of such specificity would permit in vitro synthesis of an RNA molecule having a 5'-terminal sequence identical to its wild type in vivo counterpart. To test its requirement, we replaced all of the SP6 sequence downstream of the transcription start point with heterologous nucleotides (nt) and found that any sequence will suffice to permit efficient and accurate transcription. These results permitted construction of plasmids for synthesis of 'authentic' transcripts from cloned DNA. In one case, by an oligodeoxynucleotide-mediated site-specific deletion, we placed the start point of yeast gene TCM1 at nt + 2 of the SP6 promoter and produced in vitro TCM1 mRNA with a wild-type 5'-terminal sequence. We also constructed a vector, pSP64 delta 1, in which the SalI/AccI/HincII recognition sites of pSP64 reside at nt + 2 through + 7. Plasmids such as pSP64 delta 1 may be more useful in some cases as insertion of any DNA fragment at one of these three sites will yield a transcript in which only two to four nt are derived from the vector. PMID- 3026928 TI - Selection of lambda Spi- transducing phages using the P2 old gene cloned onto a plasmid. AB - The old gene product of the P2 prophage interferes with plaque formation by lambda wild type phage but allows lambda phages whose red and gam genes have been deleted to form small, visible plaques (the lambda Spi- phenotype). The old gene product also kills Escherichia coli recB or recC mutants. We have cloned the old gene into the high-copy-number plasmid pBR322, where it prevents plaque formation by both lambda Spi+ and lambda Spi- phages. We transferred a DNA fragment that carries the old gene to the low-copy-number plasmid pSC101 and found that lambda Spi- phages can be selected on strains that carry this plasmid. The plasmid-borne old gene kills E. coli recB mutants, providing a selection for old- mutants. PMID- 3026929 TI - DNA homology between the 3'-untranslated regions of a developmentally regulated Drosophila gene and a mouse alpha-interferon gene. AB - The Drosophila genome was screened for DNA sequences that have homology with the mouse alpha-interferon gene MuIFN alpha 2, by hybridizing a library of cloned Drosophila genomic DNA with an MuIFN alpha 2 cDNA probe under reduced stringency conditions. Several dispersed regions of homology were identified, one of which was mapped at position 67D8-10 on chromosome 3. The homology in this region occurs within a gene, called ect, which encodes a 2.9-kb poly(A)+RNA transcript. The principal homology between ect and MuIFN alpha 2 involves A + T-rich sequences with short repeats, which are located within the untranslated 3' end of the transcribed region of each gene. Since no homology between the translated regions of ect and MuIFN alpha 2 was detected by hybridization, there appears to be no significant structural or functional homology between the encoded proteins. The ect gene is temporally regulated during Drosophila development, with a major peak of expression during the second half of Drosophila embryogenesis. Expression of ect occurs selectively in ectodermal tissues of the embryo, suggesting a specialized role for the gene in the early development of ectodermal structures. PMID- 3026930 TI - In vitro packaging into phage T4 particles and specific recircularization of phage lambda DNAs. AB - Concatemeric phage lambda imm434 DNA packaged in vitro into phage T4 particles produced plaques on a selective host. Moreover, lambda DNA containing a pBR322 derivative flanked by the lambda attL and attR sites could be specifically recircularized by excisive lambda recombination to yield the pBR322 derivative. A host deficient in generalized recombination and containing a defective lambda c Its prophage which provided Int and Xis proteins was the recipient for this plasmid derivative carried by T4. Such a T4-lambda hybrid may potentially allow almost one T4 headful of donor DNA (166 kb) to be packaged and recircularized. PMID- 3026931 TI - [Fiber bronchoscopic semeiotics and the morphologic diagnosis of small-cell lung cancer]. PMID- 3026932 TI - Incidence and clinical significance of colonic cytomegalovirus infection in idiopathic inflammatory bowel disease requiring colectomy. AB - Evidence of cytomegalovirus infection has been sought in a consecutive series of patients requiring colonic resection for idiopathic inflammatory bowel disease confined to the colon. Colonic tissue was examined by light and electron microscopy for cytomegalovirus inclusion bodies and cytomegalovirus antigen was sought using an immunoperoxidase technique. Cytomegalovirus infection was identified in three of 26 patients studied, but the infection did not appear to influence the course of the colitis. Although all three patients with cytomegalovirus infection needed urgent surgery, none had toxic megacolon. No evidence of cytomegalovirus infection was found in three other patients with toxic megacolon. One patient, who had a rising titre of IgG to cytomegalovirus received treatment with acyclovir which eradicated cytomegalovirus from the colon without altering the course of the colitis. PMID- 3026933 TI - Cystic papillary hidradenoma of the vulva: case report and review of the literature. AB - Papillary hidradenoma of the vulva is a rare, benign neoplasm arising from apocrine sweat glands of the skin. Frequently, this lesion has been mistaken for carcinoma. The treatment of choice is local excision. The prognosis for patients with this tumor is excellent. We present a patient who is unique because she had a lesion which was the largest ever recorded, and which existed over twice as long as any previously described. This case is also presented to remind the clinician that, despite the gross appearance of the tumor which resembles carcinoma on sectioning, biopsy and histological diagnosis should guide the ultimate management of patients with such lesions. The findings in our patient support the view that no matter how large or how long in existence, hidradenoma remains benign. PMID- 3026934 TI - Adenocarcinoma of the endometrium with trophoblastic differentiation and metastases as choriocarcinoma: a case report. AB - Nongestational choriocarcinoma is seldom observed outside of gonads or teratomas. No case of choriocarcinoma arising in endometrium is reported in the literature. Here we present a 70-year-old female with endometrial adenocarcinoma focally differentiating to choriocarcinoma, and metastasizing to liver, kidney, brain, and lung as pure choriocarcinoma. Foci of trophoblastic differentiation in the primary endometrial lesion and metastatic lesions are positive for hCG by immunocytochemical stain. This case is similar to gastrointestinal and lung carcinomas that contain trophoblastic change. Possible histogenesis of this tumor is discussed. PMID- 3026935 TI - [Estrogen replacement therapy for postmenopausal osteoporosis]. PMID- 3026936 TI - [Live attenuated varicella zoster virus (VZV) vaccine]. PMID- 3026938 TI - Detection of delta 9-THC in saliva by capillary GC/ECD after marihuana smoking. AB - A method is described for the determination of delta 9-tetrahydrocannabinol (delta 9-THC) in the saliva by the use of a combination of moving-precolumn injector and glass capillary gas chromatograph with electron capture detector (GC/ECD). There were no interfering peaks due to impurities around the peak of pentafluoropropyl derivative of delta 9-THC (delta 9-THC-PFP). This GC/ECD method was linear over the range of 5-200 ng/ml of delta 9-THC-PFP. The lower detection limit was approximately 1 ng/ml. delta 9-THC content in the saliva after experimental marihuana smoking was measured by this method. It was demonstrated that for at least 4 h after smoking the level of delta 9-THC was sufficient for detection. PMID- 3026937 TI - Hepatic responses to the administration of high doses of BHT to the rat: their relevance to hepatocarcinogenicity. AB - Although butylated hydroxytoluene (BHT) is non-mutagenic, at high doses it has recently been associated with an increased incidence of liver tumours in laboratory rodents. To establish whether chronic liver cell injury may be involved in the genesis of these tumours, BHT was administered to rats by orogastric gavage at doses of 0, 25, 250 or 500 mg/kg/day for up to 28 days and also at daily doses of 1000 and 1250 mg BHT/kg for up to 4 days (sublethal doses). The sublethal doses induced centrilobular necrosis within 48 hr, whereas administration of BHT for 7 or 28 days caused dose-related hepatomegaly and at the highest dose level induced progressive periportal hepatocyte necrosis. The periportal lesions were associated with proliferation of bile ducts, persistent fibrous and inflammatory cell reactions, hepatocyte hyperplasia and hepatocellular and nuclear hypertrophy. Biochemical changes consisted of dose related induction of epoxide hydrolase, dose-related changes in the ratio of cytochrome P-450 isoenzymes and depression of glucose-6-phosphatase. Measurement of BHT demonstrated a dose-related accumulation in fat but not in the liver. Changes in hepatic activating and detoxifying enzyme profiles are implicated both in the mechanism of periportal hepatocyte damage and in the change of site of damage according to the dose and duration of the treatment. The persistent and active nature of the lesions in rats dosed with 500 mg BHT/kg for 28 days, combined with evidence of cell damage at doses equivalent to those associated with hepatic tumours (250 mg BHT/kg), suggests that chronic liver cell damage may be involved in their aetiology. In this and several other studies, there was no evidence that BHT causes liver damage at a dose level of 25 mg/kg/day. As this is several hundred times higher than the normal human intake, it is considered unlikely that BHT poses a threat to human health. PMID- 3026939 TI - [Atrial natriuretic hormone. A factor in the pathogenesis of hypertension]. PMID- 3026940 TI - A patient with familial systemic lupus erythematosus associated with gastric cancer and a family study of HLA. PMID- 3026941 TI - Receptors for polymerized human serum albumin and other hepatitis B virus markers during acute hepatitis B--predictive value of the outcome of the disease. AB - Polymerized human serum albumin virus receptors (pHSA-R) HBsAg, HBeAg, antiHBc IgM, hepatitis B virus (HBV) DNA polymerase activity and HBV-DNA were studied in 47 acute hepatitis B patients, divided into three groups: 26 HBeAg(+) initially, with favorable outcome; 4 HBeAg (+), with chronic outcome; and 17 antiHBe (+), with favorable outcome. In the basal sample only 2 and 8 patients in Group I were HBV-DNAp and HBV-DNA positive, respectively, and became negative during the follow-up. In contrast all patients in Group II remained positive to both HBV markers. After a one-month follow-up 100% of the patients in Group II were positive for pHSA-R and HBeAg, in contrast to 25% among those with a favorable outcome in Group I (p less than 0.005). Meanwhile, only 6 out of 17 patients in Group III remained positive for pHSA-R. A significant decrease in pHSA-R and HBsAg concentrations was observed in patients from Group I (p less than 0.005 and p less than 0.05, respectively) 15 days after the onset of the disease, while concentrations of both parameters did not vary in Group II. A significant decrease in HBsAg and pHSA-R concentrations was found in patients from Group III after 15 days (p less than 0.05) and one month follow-up (p less than 0.05), respectively. As a result, pHSA-R and HBeAg are the best prognostic indicators in acute hepatitis B. A decrease in HBsAg and pHSA-R concentrations two weeks after the onset may have predictive value. PMID- 3026942 TI - What should hepatologists know about membrane fluidity? PMID- 3026943 TI - How proteins move across the endoplasmic reticulum membrane. PMID- 3026944 TI - Apocrine metaplasia in phyllodes tumours of the breast. PMID- 3026945 TI - Poorly differentiated (small cell) carcinoma of the ovary in young women: evidence supporting a germ cell origin. AB - The clinical and pathologic features, including immunohistochemistry and electron microscopy, of six cases of poorly differentiated carcinoma of the ovary (small cell carcinoma) are presented. These tumors occurred in six young patients ranging in age from 10 to 24 years. Two patients had hypercalcemia. All tumors were unilateral, and four patients had advanced stage disease at presentation. Histologic features included sheets, nests, and cords of cells in a fibrous stroma, focal microcysts, and a dimorphic population of small and large cells. Eosinophilic, hyaline globules occurred in five cases, intercellular basement membrane-like substance in two cases, and glycogen in all cases. Five of six cases stained strongly for cytokeratin and vimentin; intracytoplasmic laminin was identified in three cases; and three cases were believed to show faint positivity for alpha-1-antitrypsin. Stains for alpha-fetoprotein were negative. Ultrastructural examination of two cases showed granular material in dilated rough endoplasmic reticulum, intermediate filaments, intracytoplasmic dense globules, maculae adherens, and extracellular basement membrane-like material. All of the cases proved rapidly fatal despite various therapies, as did a histologically similar testicular tumor that was admixed with seminoma and teratoma. We interpret these findings to indicate that this ovarian cancer is most likely of germ cell origin, and it may be related to yolk sac tumor, although it is clearly distinct from the classical yolk sac tumor. PMID- 3026946 TI - Genetic linkage study between the loci for Duchenne and Becker muscular dystrophy and nine X-chromosomal DNA markers. AB - A set of nine polymorphic loci defined by DNA probes was studied for linkage with the disease locus in ten families with a history of Duchenne muscular dystrophy (DMD), and three families with a history of Becker muscular dystrophy (BMD). The results confirm DMD and BMD linkage to all marker loci and suggest closer linkage of several probes than hitherto detected. This will be of practical interest for risk calculations in affected families. PMID- 3026947 TI - The gene encoding vasoactive intestinal peptide is located on human chromosome 6p21----6qter. AB - Vasoactive intestinal peptide (VIP) is a regulatory neuropeptide involved in a wide variety of functions, among them vasodilation, smooth muscle relaxation, sweat secretion, gastrointestinal peristalsis, and pancreatic function. A deficient VIP-innervation of sweat glands was recently described as a possible pathogenic factor in sweating of cystic fibrosis (CF) patients. To investigate a possible role for a defective VIP-gene in cystic fibrosis, we have used a panel of rodent-human hybrid cells, retaining defined complements of human chromosomes to localize the VIP-gene to the human chromosome region 6p21----6qter. As the CF gene was recently mapped to chromosome 7, we conclude that the VIP-gene is not the primary gene defect in this disease. PMID- 3026948 TI - Hemoglobin M Iwate is caused by a C----T transition in codon 87 of the human alpha 1-globin gene. AB - DNA restriction, molecular cloning, and sequencing methods have been used to characterize the mutation leading to the methemoglobinemia HbM Iwate. It could be demonstrated that the HbM Iwate defect is caused by a point mutation involving a transition from C to T in the first position of codon 87 of the alpha 1-globin gene. Furthermore, the HbM Iwate mutation can directly be identified upon RsaI digestion. This direct detection of the mutation on the gene level is of significant advantage for differential diagnostic purposes. PMID- 3026949 TI - Sequences which flank an 11p deletion observed in an hepatocellular carcinoma map to 11p13. AB - There is considerable interest in the 11p13 region because of its involvement in Wilms tumor, sporadic aniridia, and other congenital abnormalities. Cloned DNA sequences from this region might be useful in understanding the chromosomal abnormalities which lead to such disorders. However, few such markers exist. Using somatic cell hybrids which contain defined 11p deletions, two cloned DNA sequences which flank a deletion generated in an hepatocellular carcinoma (as a consequence of hepatitis B virus integration) were mapped to 11p13. Thus both ends of the deletion observed in an hepatocellular carcinoma are within 11p13. PMID- 3026950 TI - An apparent discrepancy between chain length and electrophoretic mobility of restriction fragments: a case of human mitochondrial DNA. AB - DNA sequence analysis and electrophoresis in denaturing gel revealed that a 60 base pair insertion which had been previously postulated on the basis of native polyacrylamide gel electrophoresis of mitochondrial DNA from Japanese (Horai and Matsunaga 1986) did not exist at all. Unusual behavior of certain restriction fragments in native polyacrylamide gels apparently resulted in what appeared to be an insertion. Further study revealed that this behavior is most likely due to secondary structures of the fragments. The results of the present study suggest that adequate care should be taken when assessing molecular weights of restriction fragments by native polyacrylamide gel electrophoresis. PMID- 3026951 TI - DNA-polymorphism of type I collagen gene detected with Bgl II in the genetically isolated Finnish population. AB - The pro alpha 2(I) collagen gene has been screened for Bgl II restriction fragment length polymorphisms (RFLPs) in the Finnish population which has a long background of genetic isolation. As genetically isolated populations tend to demonstrate deviations in the degree of heterozygosity, the RFLP markers described in more heterogeneous populations can not be utilized as such in the former. This proved to be the case with Bgl II polymorphism within the pro alpha 2(I) collagen gene, the gene involved in different disorders of connective tissue. This report describes the presence of three new RFLP-loci and the absence of one RFLP-locus, which had been earlier reported from a population with a genetic background remote from the population studied here. PMID- 3026952 TI - Partial androgen receptor deficiency and mixed gonadal dysgenesis in Drash syndrome. AB - Drash syndrome associates a nephropathy characterized by a diffuse mesangial sclerosis of early onset, Wilms tumor, and male pseudohermaphroditism (MPH). A patient with Drash syndrome is reported with the following: karyotype 46,XY, external genitalia near normal female, mixed gonadal dysgenesis, severe androgen receptor deficiency demonstrated for the first time in this syndrome. The possibility of a common genetic denominator with the del 11p13 WAGR complex is suggested. MPH/nephroblastoma association is common. Androgen receptor deficiency has been observed in one case of each syndrome, respectively. PMID- 3026953 TI - [Limits of conventional nutrition education: an example from a West African urban area]. AB - The failure of conventional nutrition education, as practiced in Western Africa, is acknowledged by the author. This assessment follows the formative and summative evaluation of a nutrition education programme, directed towards the parents of hospitalized children, in a hospital, in the Ivory Coast. Often carried out within conventional health structures (maternal-child health centers, rural health centers, hospitals, etc...), the programme represents an addition to daily health care activities, with actions often too isolated and too limited to exert a decisive impact on public health. Most of the time, these programmes are conceived and implemented by health care workers who are inadequately motivated and trained in both nutrition and education. Poorly conceived messages that lack cultural, economic or social adaptation to the specified target population, authoritarian, unfeeling pedagogy, and inadequate educational tools lead to uncertain results. Results that are difficult to assess due to the absence of a clear definition of objectives and procedures for scientific evaluation. The author, in agreement with other recent studies, suggests that nutrition education should be viewed as the cumulative effect of all the various communication activities aimed at modifying those individual or community behaviours that affect the nutritional status of a given population. Multimedia nutrition education, however, can only be implemented once population needs and available communication channels have been identified. It is with the idea of promoting such multimedia activities that the network for nutrition education in Africa has been created (see p. 56). PMID- 3026954 TI - Organ-specific autoimmunity: a 1986 overview. AB - The normally functioning immune system is subject to intricate networks of regulatory mechanisms: it is therefore not surprising to find that autoimmune diseases present a complex pathogenic picture in which the relative contributions of various factors probably determine the precise nature and course of disease. This is particularly evident in the effector mechanisms of organ-specific autoimmunity which are described in this chapter. These ultimately give rise to the disease symptoms, and can be directly cytotoxic, or may either stimulate or block functional activity or growth of the target cells. Their various contributions to human diseases are becoming more firmly established, as in Type I diabetes, or are only now being described, as in the case of EC-Ab in protracted diarrhea of infancy and as evidenced by the growing lists of receptor stimulating or -blocking antibodies. The nature and precise location of relevant autoantigens is also coming under closer scrutiny. The answers to the question of why these diseases arise in the first place remain more elusive. However, it is again likely that a variety of factors can contribute. The attractive possibility of a role for idiotypic interactions is gaining ground, particularly within the context of antibodies to hormones and their receptors. Another potential mechanism which we believe may be of central importance, particularly in the development of organ-specific destructive autoimmunity, and which we have discussed here in detail, is the aberrant expression of HLA Class II molecules by target cells. Whether this is actually an initiating factor is presently not known, but its potential for promoting pathogenesis both early and late in the process is clear. Furthermore, the complex nature of the regulation of epithelial Class II expression may help to explain the heterogeneity of features and course of disease in different patients with the same underlying pathology. All these advances in our basic understanding of the disease processes should ultimately lead to more effective and specific means of therapeutic intervention. PMID- 3026955 TI - Noncytocidal mechanisms of action of tumor necrosis factor-alpha on human tumor cells: enhancement of HLA gene expression synergistic with interferon-gamma. AB - In this report, we present evidence that tumor necrosis factor alpha (TNF-alpha) exerts regulatory activity on the expression of HLA genes in human tumor cell lines at the level of mRNA expression and at a posttranscriptional level. These mechanisms are independent of direct cytotoxic/cytostatic effects, as enhancement of HLA antigen expression was observed in both sensitive cell lines and in cell lines resistant to TNF-mediated growth inhibition. In a priori HLA-negative cells, a strong TNF-alpha-mediated enhancement of the Interferon gamma (IFN gamma) induced expression of HLA genes was revealed by Northern blot and immunofluorescence analysis. A distinct mechanism of TNF-alpha action is suggested from enhancement of constitutively expressed HLA genes. In this case, TNF-alpha raised HLA antigen density without apparent enhancement of cytoplasmic mRNA levels. This indicates that TNF-alpha influences HLA expression also at the level of translation or at a posttranslational level. TNF-alpha treatment of HLA negative cells did not by itself result in HLA gene induction, suggesting that TNF-alpha acts as an enhancer and not as an inducer of HLA gene expression. PMID- 3026956 TI - Parallelism between superoxide production of peritoneal exudate cells and lung granulomatous response in mice vaccinated with BCG cell walls. AB - Since peritoneal macrophages are reported to be different from alveolar macrophages in their activated states, we examined whether O-2 production, one of the parameters of macrophage activation, in mouse peritoneal exudate cells (PEC) is enhanced under the condition in which lung granuloma, the accumulation of activated macrophages, is produced with Bacillus Calmette-Guerin (BCG) cell wall (CW). As a result, we observed the enhanced O-2 production of PEC that occurs in parallel with lung granuloma formation; high responders, C56BL/6 mice, showed high O-2 production of PEC whereas low responders, C3H/He and DBA/1 mice showed low O-2 production of PEC, suggesting that enhanced O-2 production of PEC as well as lung granuloma formation is genetically controlled. Results from T cell depleted mice and allogeneic bone marrow chimeric mice also showed the occurrence of this parallelism. From these findings, we presumed that circulating macrophage activating factor and other lymphokines produced by BCG CW-sensitized T cells may activate both peritoneal macrophages and lung macrophages. PMID- 3026957 TI - Functional comparison of bone marrow-derived macrophages obtained by cultivation in serum-free or serum-supplemented medium. AB - Bone marrow-derived macrophages obtained by cultivation in a serum-free or in a serum-supplemented medium were compared in terms of the activation of the respiratory burst and the activation of tumor cytotoxicity. Serum-free-cultured macrophages responded to interferon-gamma (IFN-gamma) and to lipopolysaccharide (LPS) by an enhancement of the respiratory burst. Macrophages obtained in a serum supplemented medium are characterized by a diminished capacity to release O2-. These cells did not respond to IFN-gamma unless the stimulation was performed in a serum-containing medium. In terms of activation of tumor cell cytotoxicity, serum-supplemented macrophage cultures seem to be primed by unknown serum constituents because they only need one signal (IFN-gamma or LPS) to become fully cytotoxic. Serum-free cultivated macrophages can be rendered cytotoxic only after exposure to combinations of IFN-gamma and LPS. PMID- 3026958 TI - Human peripheral blood accessory cell: isolation by hypotonic density gradient, functional, and phenotypical characterization. AB - In order to purify the human peripheral blood-derived accessory cell that cooperate with T lymphocytes in the process of mitogenic stimulation, we developed a new density gradient separation. This was based on the principle of hypotonic swelling of the cells to obtain a differential change of the buoyant densities of cells. By this method, we have obtained a highly accessory cell depleted lymphocyte fraction whose proliferative response to sodium periodate stimulation was almost aborted. Another fraction containing high accessory cell activity was further divided into Fc-receptor-positive and -negative cells. The latter revealed the highest accessory activity for T lymphocyte periodate stimulation. The cells were characterized according to a number of markers and appeared to resemble lymphoid dendritic cells. Compared with the monocyte/macrophage fraction, they showed veils and dendritiform elongations and expressed reduced values of monocyte/macrophage specific markers. Compared with the high accessory activity of these cells, monocytes/macrophages expressed a low accessory activity. PMID- 3026959 TI - Regulation of human tonsillar T-cell proliferation by the active metabolite of vitamin D3. AB - We have examined the effects of 1,25(OH)2D3 on T-cell populations isolated by buoyant density and E rosetting from human tonsils. Cell proliferation was assessed by measuring the incorporation of 125iododeoxyuridine; interleukin-2 (IL 2) production was measured using an IL-2-dependent cell line, and the number of 1,25(OH)2D3 receptors was measured by whole-cell nuclear association assay. At a concentration of 10(-7) M, 1,25(OH)2D3 inhibited mitogen-induced T-cell proliferation in all E+ T-cell populations. This effect was more pronounced in the cells from the intermediate and high density layers and was reflected both in cell proliferative responses and in relative IL-2 synthesis. By adding the 1,25(OH)2D3 during the course of the mitogen assay, we demonstrated that activation of the T cell precedes the 1,25(OH)2D3-mediated inhibition. Cells that had been preincubated with mitogen in the presence of the 1,25(OH)2D3 were refractory to further stimulation by mitogens. Receptors for 1,25(OH)2D3 could not be detected in unstimulated T cells. However, activation led to the expression of high-affinity receptors for 1,25(OH)2D3. Co-incubation of the cells with mitogen and 1,25(OH)2D3 increased the number of receptors compared with mitogen alone. The effects provide further evidence for the hypothesis that 1,25(OH)2D3 is an important potential modulator of the immune system through its action on T cells. Taking our observations in conjunction with the known capacity of monocytes to hydroxylate the precursor metabolite (and thus synthesize the active form of cholecalciferol), the results support the suggestion that 1,25(OH)2D3 plays a role as a local mediator of mononuclear phagocyte-T cell interaction in human lymphomedullary tissues. PMID- 3026961 TI - Leukotriene B4 induces enhanced migration of fish leucocytes in vitro. AB - Leukotriene B4 (LTB4) was found to induce enhanced migration of the eosinophilic G1 granulocyte of the dogfish Scyliorhinus canicula in the migration under agarose assay. Higher levels of LTB4, however, were required to produce this effect than with mammalian neutrophils under similar conditions. It is postulated that this may be due to the dogfish granulocytes possessing fewer receptors for LTB4 than their mammalian counterparts. The eosinophilic G3 granulocyte was also tested using the same assay but results were inconclusive. The effect of LTB4 on dogfish G1 and G3 granulocytes was also monitored with the bipolar shape formation (BSF) assay. LTB4 induced BSF in both granulocyte types, and this method appeared to be more sensitive than the migration under agarose assay. Whether the enhanced migration observed is a result of chemotaxis or chemokinesis is not determined. This present study represents the first known report of the function of LTB4 in a non-mammalian vertebrate. PMID- 3026960 TI - Silica decreases phagocytosis and bactericidal activity of both macrophages and neutrophils in vitro. AB - Silica, or silicon dioxide, has been shown to be toxic for macrophages. This is probably because it damages phagolysosomal membranes, allowing lysosomal enzymes to disrupt the cell. Neutrophils also take up particles such as silica and in addition they contain lysosomes. The purpose of this study was to determine whether incubation in vitro with silica inhibits function not only of mouse macrophages, but also of mouse neutrophils. The data show that incubation with silica for 1-3 hr decreases viability of both macrophages and neutrophils. Silica decreases the ability of macrophages and neutrophils to phagocytose both erythrocytes and bacteria, and it inhibits the ability of both cells populations to kill the facultative intracellular bacterium Listeria monocytogenes. Thus, it appears that silica, at least in vitro, is harmful to neutrophils as well as to macrophages. PMID- 3026963 TI - Different inducing activities for cytotoxic T cells by heterogeneous thymocyte activating factors from SV40-transformed human embryo fibroblasts. AB - Thymocyte-activating factors are produced by human embryo fibroblasts and their production is enhanced by SV40-induced transformation (Okai, Gotoh and Yamashita, Immununol. Lett. (1985), 9, 153-159). The activities for thymocyte DNA synthesis in the culture medium from SV40-transformed cells are separated into the two fractions by a DEAE Sephadex A-25 column chromatography eluting at 150 and 500 mM NaCl. Each fraction contains the heterogeneous molecular weight activities as judged by a Sephadex G-100 column chromatography. In the fraction eluted at 150 mM NaCl, the inducing activities for cytotoxic T cells by the factors are dependent upon their molecular weights; the larger factors exhibit much higher cytotoxic activity than that of the smaller factors. In contrast, the heterogeneous molecular weight factors eluted at 500 mM NaCl do not show the remarkable difference of their biological activities. In addition, the inducing activities by thymocyte-activating factors are also dependent upon their charge properties. These different inducing activities for cytotoxic T cells by thymocyte-activating factors is discussed from the aspect of the immunological regulation. PMID- 3026962 TI - MHC class II antigen and immunoglobulin expression in spontaneous phenotypic variants of the Burkitt's lymphoma cell line Namalwa. AB - Phenotypic variant sublines of the Burkitt's lymphoma cell line Namalwa were examined with cDNA probes for the different MHC class II beta chain genes and with monoclonal antibodies specific for the corresponding cell surface antigens (DP, DQ and DR antigens). Expression of MHC class II antigens in the Namalwa sublines (known as CSN/70, IPN/45, PNT and KN2) was compared with that of the B lymphoblastoid cell line DEW1, which is identical to Namalwa in DR allotype (DR 2,4). There were markedly different levels of expression of MHC class II antigens among the cell lines: in DEW1 and the Namalwa KN2 subline DP, DQ and DR antigens were expressed on almost all the cells. On the PNT and IPN/45 sublines, DR antigens were expressed on all the cells, and DP and DQ antigens were expressed at detectable levels on only a proportion of cells. On CSN/70, there was weak expression of DR antigens on a minority of cells and no detectable expression of DP and DQ antigens. When examined with MHC class II-specific cDNAs, restriction fragment patterns of DNA were identical for all the cell lines, suggesting that they had structurally identical MHC class II genes. In the Namalwa cell lines the synthesis of Ig and the expression of MHC class II antigens were coordinately regulated. PMID- 3026964 TI - Age-related changes of beta-adrenoceptors in spleen lymphocytes and cerebral cortex of NZB/BIN mice. AB - The density (Bmax) of beta-adrenoceptors in splenic lymphocytes of NZB/BIN mice decreased up to an age of about 40 weeks and then levelled out. The Bmax in cerebral cortex, on the other hand, increased in the first half of life and then changed relatively little. The dissociation constant of the ligand (Kd) was larger in the cortex than the spleen and showed relatively little age-dependent change. PMID- 3026965 TI - Cooperative action of complement component C3 and phagocytic effector cells in innate murine resistance to Trypanosoma lewisi. AB - Mice display strong natural resistance to the rat-specific Trypanosoma lewisi. Intravenously injected intact T. lewisi parasites were eliminated by mice within 18 to 24 h. In comparison, "nude" T. lewisi organisms lacking the surface coat were rapidly and totally cleared from the bloodstream within 2 h postinoculation. Similarly, in vitro-cultivated trypanosomes were readily eliminated by mice with the conspicuous absence of a lag phase. Elimination of nude T. lewisi, like that of intact trypanosomes, required murine complement component C3. Splenectomy of mice did not affect their ability to eliminate T. lewisi. However, C3 depletion with cobra venom factor rendered splenectomized mice susceptible to this rat trypanosome; in these mice, T. lewisi established prolonged and frequently fatal infections. Beige mice were able to efficiently eliminate T. lewisi. But combined treatments of beige (bg/bg) and heterozygous (bg/+) mice with cobra venom factor and silica dust, or normal rat serum and silica dust, incapacitated the natural resistance of these mice to T. lewisi. Such combined treatments of beige and control mice resulted in fulminating parasitemias and death of the animals. Altogether, the results of the present studies indicate that T. lewisi elimination by mice requires the following: exposure of C3 acceptors on the surface of the parasites; activation of murine C3, probably via the alternative complement pathway; and destruction of the C3b-coated parasites by their interaction with C3b receptor-bearing, phagocytic effector cells that are abundant in the spleen and sensitive to silica dust. PMID- 3026966 TI - "Haemophilus somnus," a facultative intracellular pathogen of bovine mononuclear phagocytes. AB - We have reported previously that bovine neutrophils are unable to kill the bovine respiratory pathogen "Haemophilus somnus." In the present study we expanded our efforts and examined the interaction of bovine mononuclear phagocytes with this important veterinary pathogen. Bovine alveolar macrophages and blood monocytes ingested but did not kill opsonized "Haemophilus somnus" in vitro, whereas these same cells ingested and killed opsonized Escherichia coli. Because this suggested that "H. somnus" was a facultative intracellular pathogen, we developed an assay to monitor the intracellular fate of ingested "H. somnus" within bovine monocytes. Our results indicated that ingested "H. somnus" multiplied within bovine monocytes (1- to 2-log10 increase in 4 h); equivalent intracellular growth was noted for both a laboratory strain and a recent field isolate of "H. somnus." Bovine monocytes killed ingested E. coli (1- to 2-log10 decrease in 4 h) under the same assay conditions that were used to follow intracellular growth of "H. somnus," thus indicating that the assay conditions did not induce a generalized defect in monocyte antibacterial activity. Light and electron microscopic examination of "H. somnus"-infected monocytes confirmed that intracellular growth had occurred. We did not observe an obvious correlation between the release of superoxide anion from bovine mononuclear phagocytes that had ingested opsonized "H. somnus" and E. coli and the subsequent intracellular survival of the bacteria. The results of this study suggest that infected mononuclear phagocytes sustain "H. somnus" infections in cattle and thus contribute to the subacute to chronic clinical course that has been reported. PMID- 3026967 TI - Activity of nedocromil sodium in mast-cell-dependent reactions in the rat. AB - Nedocromil sodium, a pyranoquinoline dicarboxylic acid derivative which differs structurally and physiochemically from sodium cromoglycate, was nonetheless shown to be effective in classical models of anti-allergic activity in the rat. These models, based on immediate hypersensitivity reactions in the passively sensitized rat, involve release of mediators from mast cells following cross-linking of membrane-bound IgE antibodies by specific antigen. In this system, nedocromil sodium exhibited a similar profile of activity to that of sodium cromoglycate. Whilst not predictive for therapeutic activity, rat models of immediate hypersensitivity are believed to reflect one component of obstructive airways disease. The results suggest that nedocromil sodium is worthy of clinical evaluation in this indication. PMID- 3026968 TI - Maternal cannabinoid exposure. Effects on spermatogenesis in male offspring. AB - Maternal exposure to cannabinoids influenced spermatogenesis and fertility in their male offspring examined at 60-80 days of age. Approximately 20% less spermatozoa were found in males whose mothers had received either the non psychoactive cannabinol (CBN) or cannabidiol (CBD) on day 1 postpartum. Males exposed to the major psychoactive component of marihuana, delta 9 tetrahydrocannabinol (THC) appeared to have spermatozoa in number comparable to controls. This finding may be consistent with the additional observation that CBN or CBD, but not THC, reduced the percentage of successful impregnations by cannabinoid-exposed males. However, males exposed to each of these cannabinoids produced significantly less live offspring compared to controls. Plasma levels of testosterone and luteinizing hormone (LH) were reduced significantly in mice exposed to THC on day 12 of gestation, while testicular weight was reduced in adult mice exposed either on day 12 of gestation to CBD or on day 1 post-partum to THC. These results indicate that perinatal exposure to psychoactive and non psychoactive components of marihuana can produce long-term disruption of testicular function including the spermatogenic as well as the steroidogenic components. PMID- 3026969 TI - Pretreatment prognostic factors and scoring system in 407 small-cell lung cancer patients. AB - In 407 patients with small-cell lung cancer (SCLC), 61 pretreatment variables were evaluated in a Cox multiple regression analysis to assess their prognostic value. All patients received short-term intensive regimens (cyclophosphamide, etoposide and methotrexate or ifosfamide and etoposide, both followed by thoracic irradiation if complete response was noted). Lactate dehydrogenase (p = 0.001), tumour stage (p = 0.0001), serum sodium (p = 0.0009), pretreatment Karnofsky performance score (p = 0.0121), alkaline phosphatase (p = 0.0186) and serum bicarbonate (p = 0.0321) were the important prognostic factors. Once these variables were taken into account no other variable provided additional prognostic information. A simple scoring system ("Manchester Score") using these variables was established and shows little loss of information compared to the Cox analysis. The score distinguishes 3 prognostic groups, the best of which contains all long-term survivors, whereas the bad prognostic group contains no patient surviving longer than one year. The scoring system may help to design new treatment strategies and may also facilitate the comparison of different studies. PMID- 3026970 TI - Progenitor and pre-B lymphocytes transformed by Epstein-Barr virus. AB - By rosetting with SRBC coupled to rabbit-anti-human IgM, the surface IgM-negative cells of human fetal bone marrow were enriched, and subsequently infected and transformed by Epstein-Barr virus (EBV). Single clones of the transformed cells were obtained. Ninety percent of the resulting cell clones were surface immunoglobulin-negative, and of 8 clones which were further studied, 5 lacked intracellular, cytoplasmic Ig as measured by immunofluorescence. Control cell clones derived from the same material without pre-selection expressed surface Ig and also secreted Ig. Utilization of a panel of B-cell-specific monoclonal antibodies (MAbs) showed no difference between the cell clones expressing surface Ig and those that did not. The progenitor B-cell lines did not show a phenotype resembling that of cell lines derived from B-cell malignancies, such as high agarose clonability. In spite of their immature Ig-phenotype, these clones showed rearrangement of at least one heavy chain Ig-allele. Efforts to induce differentiation in these clones were unsuccessful. These clones may represent progenitor B cells, or B cells with faulty heavy-chain rearrangement. EBV can apparently be used as a tool to derive cell lines representing different levels of B-cell differentiation, and can also transform immature B cells, which may be useful in the analysis of B-cell differentiation. PMID- 3026971 TI - HTLV-III infection of EBV-genome-positive B-lymphoid cells with or without detectable T4 antigens. AB - The infection of a number of new and established B-cell lines by human T-cell lymphotropic virus III (HTLV-III) was investigated. The B lymphocytes differed in their expression of T4 antigens detected by specific monoclonal antibodies (MAbs) and the presence of Epstein Barr virus (EBV)-DNA or antigens. The presence of the EBV genome was the only requirement for infection of B-lymphocytes by HTLV-III, although its presence did not ensure infection. Two EBV genome and T4 antigen positive B-cell lines, lacking EBV early antigens (EA) and viral capsid antigens (VCA), could be productively infected with no induction of known EBV antigens. Two other EBV genome-positive cell lines, lacking T4, EA, and VCA could also be infected. Another genome-positive cell line (P3HR-I) that was EBV-EA, VCA positive and produced non-transforming EBV, could also be infected by HTLV-III. However, 3 EBV genome- and T4 antigen-negative B-cell lines could only be infected with HTLV-III after successful conversion to an EBV-genome-positive state by pre-infection with EBV. Five other EBV-genome-positive B-cell lines lacking T4 antigens were not infectible with HTLV-III even after super-infection with EBV. Incomplete inhibition of the HTLV-III infection of a T4-positive (LDV 7) and a T4-negative (Craig) was obtained by preadsorption with specific MAb to T4 (OKT4A and Leu 3A). From these observations, it is not clear whether the presence of T4 antigen on the cell surface is needed for the infection of B lymphoblastoid cells; however, successful infection does depend upon the presence of the EBV genome. The mechanism of interaction of HTLV-III and EBV-infected B cell lines permitting this infection is not fully understood. Although the clinical implications of these observations remain to be determined, it is possible that infection of EBV-positive B-cells may contribute to aberrant humoral responses and/or increased frequency of B-cell malignancies observed in HTLV-III-infected individuals. PMID- 3026972 TI - Enhancement of production of superoxide anion by human monocytes exposed to products of HT 29 human colonic adenocarcinoma cell line. AB - The generation of superoxide anion (O2-) by human blood monocytes in response to stimulation by either phorbol myristate acetate (PMA) or opsonized Zymosan was greatly enhanced (range: 100-200% according to donor) by prior exposure of the peripheral blood mononuclear cells (PBM) to human colonic adenocarcinoma cells (HT 29 line) or their conditioned culture medium (DMEM-HT 29). This priming effect was observed after 5 hr and persisted for up to 15 hr of contact between PBM and endotoxin-free DMEM-HT 29. Beyond this time, primed monocytes gradually lost this ability. However, they maintained a higher capacity (about 100%) to produce O2- when compared to controls. DMEM-HT 29-induced monocyte priming requires that the tumor-active substance(s) act(s) on 2 target cells: first, on non adherent mononuclear cells (NA-PBM) to induce cytokine production and, second, on the monocyte itself. Priming activity was also found in conditioned medium from FR3T3 embryonic fibroblasts but not in conditioned medium from HT 29 repolarized cells (by culture in glucose-free medium) or from non-tumorous human colonic mucosa explants. PMID- 3026973 TI - Expression of B-cell-specific markers in different Burkitt lymphoma subgroups. AB - Forty-three Burkitt lymphoma (BL) lines were examined for the expression of 5 monoclonal antibody (MAb)-identified B-cell-specific markers and immunoglobulin production. All (13) EBV-negative BL lines were CALLA+ LB-1-, whereas 30 EBV carrying lines showed a more heterogeneous pattern. In the EBV-negative lines, the follicle mantle zone markers BA-1 and 35.1C5 were expressed concordantly, at a different level in each line. This coordination was disrupted in EBV-carrying lines. In the EBV-negative lines, there was also an inverted correlation between the expression of 35.1C5 and the germinal center marker BLA, suggesting that some etiologically important event, perhaps the translocation, had fixed the cells at different stages of their transition from one zone to the other. This inverted relationship was also disrupted in the EBV-carrying lines, suggesting that EBV can interfere with the maturation program of the BL cell. This conclusion was also supported by a comparison between 5 EBV-negative BL lines and their EBV converted sublines. All converted lines have undergone marker changes, but the degree and nature of these changes was different for each EBV-BL line. Both the coordinated expression of BA-1 and 35.1C5 and the inverted relationship between CALLA and LB-1 were disrupted in several other convertants. We have reexamined our previous finding (Ehlin-Henriksson and Klein, 1984) that the majority of the variant translocation-carrying BL lines were CALLA- LB-1+, in contrast to the majority of the typical translocation carriers that were mostly CALLA+ LB-1-. All II EBV-negative lines were CALLA+ LB-1-, irrespective of the type of translocation. Among the EBV-carrying lines, 4 of 17 typical (8;14) translocation carriers were CALLA- LB-1+, whereas 7 of the 12 variant translocation-carrying lines were CALLA- LB-1+. The remaining two expressed both antigens to some extent. The difference is statistically significant at the 0.03 level. PMID- 3026974 TI - In vitro effects of sodium diethyldithiocarbamate (Imuthiol) on human T lymphocytes. AB - The effect of purified diethyldithiocarbamate, DTC (Imuthiol) on human T-cell dependent functions has been investigated. The mitogenic response of PHA stimulated peripheral blood lymphocytes (PBL) was significantly enhanced by Imuthiol at low drug concentration (10(-7) mg/ml). The same dose of Imuthiol also stimulated IL2 production by human PBL. This enhancement depended on the presence of adherent cells. These results could, in part, explained the immunostimulant activity of Imuthiol. PMID- 3026975 TI - Fate and distribution of radioactive sodium diethyldithiocarbamate (Imuthiol) in the mouse. AB - The distribution of 35S in mouse tissues has been investigated by radioactivity counts and by autoradiography after the intravenous injection of 35S-labeled imuthiol (sodium diethyldithiocarbamate). Radioactive Imuthiol is selectively localized on liver, thymus and brain neocortex, most likely as the methyl ester, within minutes after dosing. Lung or white brain matter did not fix the labeled thiol. Blood, kidney and guts show rapid elimination patterns, and can be considered as passage organs which did not fix Imuthiol. The findings are consistent with the immunopharmacological data which demonstrate that Imuthiol exerts its T-cell recruiting and activating influence through a multi-step pathway involving the brain neocortex, the thymus and the liver. PMID- 3026976 TI - Oriental sore. A look at trends in and approaches to the treatment of leishmaniasis. PMID- 3026977 TI - Atypical fibrous histiocytoma of the tongue. A case report. PMID- 3026978 TI - Human papillomavirus type 16 infection: a morphological spectrum with evidence for late gene expression. AB - Human papillomavirus (HPV) type 16 is a unique strain found almost exclusively in squamous precancerous lesions and invasive carcinomas of the genital tract. A histological and immunohistochemical analysis of 34 cervical biopsies from which HPV 16 was isolated was performed to determine: the morphological spectrum of HPV 16 "infection" and if HPV late genes were expressed in lesions containing this virus. Twenty-eight of thirty-four (82%) biopsy specimens contained areas fulfilling the histological criteria for cervical intraepithelial neoplasia (CIN) (defined as the presence of abnormal mitoses and/or diffuse nuclear atypia in a portion of the biopsy). However, 19 of 28 (68%) CIN lesions also contained focal areas of epithelium indistinguishable from condylomata. Three biopsy specimens each contained condyloma only and normal-appearing squamous epithelium. Three of thirty-one lesions (10%) showed evidence of HPV capsid antigens. The presence of areas of condyloma, as well as capsid antigens, indicates that lesions containing HPV 16 share certain similarities with conventional warts associated with other HPVs. Furthermore, the wide range of morphology in lesions containing HPV 16 suggests that histological or cytological recognition of HPV 16-associated CIN lesions may not always be possible. PMID- 3026980 TI - Mixed mesodermal tumor of the uterus in a 4-year-old girl. AB - A case of malignant mixed mesodermal tumor (MMT) of the uterus in a 4-year-old girl is reported. The patient had a polypoid lesion protruding from the vagina which was initially thought to be a sarcoma botryoides. This is one of the youngest cases of MMT on record. A review of MMT in premenopausal women and children is presented. PMID- 3026979 TI - Sensitivity of koilocytosis, immunocytochemistry, and electron microscopy as compared to DNA hybridization in detecting human papillomavirus in cervical and vaginal condyloma and intraepithelial neoplasia. AB - The sensitivity in detecting human papillomavirus (HPV) by histological observation of koilocytosis, immunocytochemistry, and electron microscopy with reference to the results of Southern blot deoxyribonucleic acid (DNA) hybridization were reviewed in 41 lesions (37 patients) of cervical and vaginal condylomata acuminata and intraepithelial neoplasia. Human papillomavirus DNA was demonstrated in fresh tissues by Southern blot DNA hybridization in all but one lesion of moderate dysplasia (98%). The rate of koilocytosis observed in tissue sections was 80% in condyloma, and ranged from 89-20% in cervical intraepithelial neoplasia (CIN), with steady reduction as the grade of CIN or vaginal intraepithelial neoplasia (VaIN) was higher. The immunocytochemistry for HPV capsid antigens was positive in 80% of condylomata and ranged from 61-0% in CIN or VaIN. The rate declined in inverse proportion to the grade of CIN or VaIN. Electron microscopy of preselected areas containing intranuclear inclusions in paraffin sections of 10 lesions demonstrated HPV-like particles in 90% of the lesions. Although immunocytochemistry and observation of koilocytosis may be useful in detecting HPV in condylomata acuminata and mild dysplasia, their sensitivity was poor in CIN or VaIN of higher grades. Electron microscopy on preselected areas in paraffin blocks showed better sensitivity, presumably due to its ability to detect immature virions. PMID- 3026981 TI - Psychosocial adjustment of familial polyposis patients and participation in a chemoprevention trial. AB - Psychological and social adjustment was assessed in eighty-nine individuals with familial polyposis, a genetically transmitted disease placing one at high risk for colon cancer. Three illness-related concerns were identified: fear about future health due to the high risk for cancer; guilt about transmitting a genetic disease to one's children; and concern about physical disfigurement resulting from surgery. Well-being scores were generally positive, although somewhat lower than those reported in a community sample. Two factors in particular influenced well-being scores: those with higher levels of concern about disfigurement reported lower well-being, and those with accurate information about the disease reported higher well-being. Of the eighty-nine individuals included in this study, sixty-one were participating in a clinical trial and twenty-eight had been invited but declined entry. Demographic and psychosocial factors were examined for their relationship to participation. Only three of these variables, length of time since diagnosis, religious affiliation, and geographic location distinguished participants from nonparticipants. PMID- 3026983 TI - Hydroxyapatite implantation--clinical and histologic analysis of a treated lesion and speculations regarding healing phenomena. PMID- 3026982 TI - Effects of gamma-irradiation on the erythrocyte membrane: ESR, NMR and biochemical studies. AB - The effects of gamma-irradiation on resealed erythrocyte ghosts have been examined with different techniques. Phospholipid analysis reveals peroxidative damage on the polyunsaturated chains of phosphatidylethanolamine. Gel electrophoresis and ESR measurements indicate modifications of the cytoskeletal proteins. 31P Nuclear magnetic resonance data show bilayer modifications that can be interpreted as changes in lipid-protein interactions. The overall picture from the present results favours interaction between lipids and proteins in the inner monolayer of the membrane. PMID- 3026984 TI - The retinal nerve fiber layer. PMID- 3026985 TI - Different kinds of acetylcholine release from the motor nerve. PMID- 3026986 TI - Photoreceptor signals and vision. Proctor lecture. AB - In recent years, there has been rapid progress in understanding the properties and mechanism of generation of the light-evoked electrical signals of vertebrate rods and cones. The graded hyperpolarization that carries information over the length of the cell is generated by closure of cation-selective aqueous pores in the surface membrane of the outer segment. These pores are controlled cooperatively by cyclic GMP, which acts continuously in darkness to keep the pores open. Photoisomerization of rhodopsin or cone pigment produces the rapid amplified activation of phosphodiesterase, which lowers the concentration of cGMP, thereby lowering the conductance of the surface membrane. Calcium ions, once thought to relay excitation to the light-sensitive channels, do not play this role. Instead, they appear to participate in a feedback control mechanism that regulates the nucleotide cascade. Although some general features of the transduction mechanism are now understood, a number of important questions remain. How is the nucleotide cascade shut off? Where does Ca act? What is the structure of the light-sensitive channel? How are stereotyped single photon responses produced? Primate photoreceptors are no longer off limits to single cell electrophysiology. Analysis of the response properties and dark noise of primate rods gives a physiological basis for several fundamental features of human rod vision: single photon detection, poor temporal resolution, the "dark light," rod saturation, scotopic spectral sensitivity, and, perhaps, after-image signals. Primate cones show less sensitive but faster responses shaped by a resonance which may figure in the flicker sensitivity of human cone vision. The spectral sensitivity of the three types of primate cones has been determined over the entire visible region. These sensitivities satisfactorily predict human color matching. The spectral sensitivity curves indicate that the pigment in a given cone is very pure, and that individual cones of a given type normally contain pigments with very similar or identical spectral properties. PMID- 3026987 TI - Gene expression and genetic engineering in the lens. Friedenwald lecture. PMID- 3026988 TI - Malignant myxoid emboli in a patient with a primary tumor of the aorta. PMID- 3026989 TI - Common colds. PMID- 3026990 TI - Replication of Aleutian disease virus in mink lymphocytes infected in vitro. AB - The kinetics of Aleutian disease virus (ADV) replication in mink lymphocytes was followed by the analysis of virus-specific antigens, infectious virus, and viral DNA. Stimulation with pokeweed mitogen (PWM; 20 micrograms/ml) increased the synthesis of ADV DNA in cultivated cells of the B-cell fraction. Maximum virus titers [10(6.5) fluorescence-forming units (FFU)/ml] were achieved after incubating infected cells for 60 h at 32 degrees. At this time, 10.4% of the cells in the B-cell-enriched fraction contained ADV-specific antigens and there was an average of 125 ADV genome equivalents per antigen-positive cell. ADV replication also was detected in T-cell-enriched fractions (with up to 10(5.3) FFU/ml, 5.3% of cells were antigen-positive, with 20 ADV genome equivalents per antigen-positive cell), but was 10 times lower compared with ADV replication in the B-cell fraction. PMID- 3026991 TI - Induction of Junin virus persistence in adult athymic mice. AB - To determine the role of T lymphocytes in adult mice infected with Junin virus, 60-day-old athymic (nu/nu) mice and their immunocompetent (nu/+) littermates were inoculated intracerebrally with 10(3) TCD50 of the XJ strain. None of them exhibited neurologic illness during a 6-month observation period, and mortality was 3% for nu/nu and 7% for nu/+ animals. The main features in infected nu/nu mice were: high viral titers in brain, reaching a late peak (6.5 log/ml) 32 days postinoculation and persisting at least 6 months; low, late viremia appearing simultaneously with the viral peak in the central nervous system (CNS) and persisting up to 3 months after infection; absence of signs of neurologic disease or histologic lesions in brain and almost no mortality; and lack of detectable circulating antibodies either in IgM or in other immunoglobulins (Igs). Circulating anti-Junin antibodies were demonstrated in IgM and other Igs in immunocompetent mice, although no infectious virus or histologic lesions could be detected. These results show an important role for T lymphocytes in the clearance of Junin virus in the CNS, as demonstrated by viral persistence induced in adult athymic mice. PMID- 3026992 TI - Modification of Junin virus neurotropism in mice by selective brain or spinal cord passaging. AB - The percentage of suckling mice that developed paralysis after intracerebral Junin virus (XJ-JV pathogenic strain) inoculation (13.8%) consistently increased after 5 serial passages of virus-infected brain or spinal cord obtained from paralytic animals, reaching 37.9 and 45.7%, respectively. As expected, all paralytic mice exhibited an identical spinal cord histologic picture, with widespread JV antigen in spinal cord astrocytes and neurons, particularly the large motor neurons of the anterior horn. These findings strongly support the existence of a motor neurotropic viral particle subpopulation in parental XJ-JV stock. PMID- 3026993 TI - Bilateral familial breast cancer. PMID- 3026994 TI - Differential transcriptional pattern of the hemopexin gene. AB - In this paper we have analyzed the expression of the human hemopexin gene in different human tissues and cell lines by using the specific cDNA probe previously isolated. The results show that this gene is expressed in liver and, in lower amount, in hepatoma cell lines but not in kidney, spleen, placental cells, and in HeLa, fibroblast cell lines. We suggest that there must be cell specific control mechanisms responsible for the specificity of expression. We have also determined, by S1 mapping, that the transcription initiation site in hepatic cells is 28 base pairs upstream from the AUG initiation codon of the hemopexin gene. PMID- 3026995 TI - Participation of protein kinase C in the activation of human neutrophils by phorbol ester. AB - The calmodulin antagonist N(6-aminohexyl)-5-chloro-1-naphthalene-sulfonamide (W 7) has been examined as an inhibitor of superoxide anion production and granule exocytosis in phorbol ester (PMA)-activated neutrophils. Inhibition of the respiratory burst was observed at a concentration of W-7 identical to that required for inhibition of native protein kinase C (PKC), whereas the concentration required to inhibit the secretory response was found to correspond to that required for inhibition of the proteolytically converted fully active PKC. The IC50 of W-7 was in both cases 5 and 12 fold higher than that required for inhibition of calmodulin dependent kinases. The results confirm the essential role for the membrane-bound PKC in the production of O2- radicals and provide a clear evidence of the direct participation of the proteolytically activated cytosolic PKC to the secretory response of PMA activated neutrophils. PMID- 3026996 TI - Dementia-peripheral neuropathy during combined deficiency of vitamin B12 and folate. Light microscopy and ultrastructural study of sural nerve. AB - We present a case of a 56 year old woman who, in consequence of repeated surgical resections of the small bowel, developed a slowly progressive and mainly motor polyneuropathy affecting the lower limbs. The polyneuropathy due to a combined deficiency of vitamin B12 and folate, was associated with intellectual deterioration, involving especially the mnesic functions. Light microscopy and ultrastructural study of sural nerve revealed: marked reduction of every order of myelinated fibers: aspects of wallerian-like myelin degeneration; various stages of myelin leaflet delamination into the cytoplasm of Schwann cells and/or macrophages; lack of alterations attributable to a primary axonal degeneration. The morphological results suggest a primary demyelination, followed secondarily by either axonal changes or some aspects of wallerian-like degeneration. PMID- 3026997 TI - Peripheral nerve involvement in chronic liver disease. Clinical and electrophysiological study. AB - A clinical and electrophysiological study was carried out on 19 selected patients with chronic liver disease. Clinical signs of peripheral nerve involvement were found in 4 patients (21%); while electrophysiological impairment was present in 11 patients (57.8%). These abnormalities were mostly limited to the sensory and motor fibers of the tibialis posterior nerve. Our data confirm the presence of peripheral nerve involvement in chronic liver disease, and that it may be evidenced by careful electrophysiological examination. PMID- 3026998 TI - [Urogenital and anal papillomavirus infections]. AB - Recently virologists and clinicians have focused attention on infections with human papillomaviruses (HPV). This is due to the ubiquity, the increasing frequency and the possible association of these viruses with the development of squamous cell carcinomas of the skin and of the mucous membranes of the respiratory, gastrointestinal, genitourinary and anorectal tracts. HPV represent a very heterogeneous group of DNA tumor viruses. By means of molecular-biological techniques, more than 40 HPV types have been recognized. In the urogenital and anal tract, papillomaviruses have been associated with venereal warts (condylomata acuminata), which have been known and recognized as a sexually transmitted disease since the Romans. Furthermore, an association has been made recently between HPV and nonpapillomatous, sometimes macular lesions: flat condylomata of the uterine cervix and of the vagina, flat condylomatous lesions and pigmented papules. The latter are localized at the mucocutaneous borders and at the skin of the lower genital tract and of the perianal and crural region. Like epidermodysplasia verruciformis, only some virus types (HPV 16, HPV 18) are regularly found in malignant, invasive squamous cell carcinomas of the genital tract, whereas others (HPV 6, HPV 11, HPV 2, HPV 10, HPV 31) are associated preferentially with benign papillomas and dysplasias. In view of the different possible oncogenic potential of the individual genotypes, early determination of the virus type probably has not only diagnostic but also prognostic value. As HPV 16 DNA is regularly present in bowenoid papulosis (flat condylomatous lesions and pigmented papules of the male genital tract), a natural reservoir has been found from which these viruses could be transmitted to the sexual partner. Knowledge of the HPV-associated clinical pictures is therefore the prerequisite for diagnosis and treatment of both the patient and his sexual partner. Clinical observation, cytology and virus typing from genital smears of both partners represent preventive methods that may contribute to the early detection of genital cancer. PMID- 3026999 TI - Determination of plutonium in feces and urine. AB - A method was developed for the determination of plutonium (239Pu, 238Pu, etc.) in feces and urine excreted by persons for 24 h. Plutonium is successively separated from inorganic macrocomponents present in samples and from other alpha emitters by the help of coprecipitation with BiPO4, LaF3 and extraction with TTA. After separation, the electrodeposition of plutonium on stainless-steel discs can be carried out. The chemical yields of separation of plutonium are about 80%. PMID- 3027000 TI - Preparation of 11C-labelled SCH 23390 for the in vivo study of dopamine D-1 receptors using positron emission tomography. AB - The dopamine D-1 receptor antagonist, SCH 23390 ((R)-(+)-8-chloro-2,3,4,5 tetrahydro-3-methyl-5-phenyl-1H-3- benzazepin-7-ol), was labelled by alkylation of the desmethyl compound SCH 24518 ((R)-(+)-8-chloro-2,3,4,5-tetrahydro-5-phenyl 1H-3-benzazepin-7- ol) with [11C]methyl iodide. A multivariate optimization method, Simplex, was employed to obtain the optimal radiochemical yield. Both straight-phase and reversed-phase preparative HPLC were investigated in the purification of [11C]SCH 23390. Reaction in acetone with subsequent straight phase LC separation resulted in 80% radiochemical yield, based on [11C]methyl iodide, with a total synthesis time of 35-40 min and a radiochemical purity greater than 99%. The average specific activity was on the order of 11.1 GBq/mmol. The 11C-labelled SCH 23390 was used to visualize the dopamine D-1 receptor-rich areas of a monkey brain by positron emission tomography. The data obtained showed a rapid distribution of radioactivity into the brain and a conspicuous accumulation of [11C]SCH 23390 in the striatum. PMID- 3027001 TI - Modular automation in PET tracer manufacturing: application of an autosynthesizer to the production of 2-deoxy-2-[18F]fluoro-D-glucose. AB - A compact autosynthesizer was developed and used successfully for the production of 2-deoxy-2-[18F]fluoro-D-glucose [18FDG] from gaseous acetyl hypo[18F]fluorite. The autosynthesizer performs a sequence of general purpose synthesis procedures named Synthesis Unit Operations (SUO's). Each SUO is controlled through execution of a digital control algorithm with a BASIC language subroutine. This automatic synthesis system is based on two industry standard microcomputer architectures, the IBM PC and STD Bus, and it becomes a component of an evolving distributed microprocessor network of task-dedicated subsystems suitable for automated manufacturing of several useful radiotracers. The yield of 18FDG product using the autosynthesizer and remote manually controlled purification procedures is approximately 20% EOB. Radiochemical purity of this product as measured by thin layer chromatography was 96-99%. Chemical purity of the product was measured to be approximately 96%. 2-Deoxy-2-fluoro-D-mannose impurity from this method was determined to be approximately 4%. PMID- 3027002 TI - An automated high pressure reaction vessel for routine preparation of short-lived radiopharmaceuticals. AB - A remote, automated high-pressure/temperature system for preparation of radiopharmaceuticals is described. The system was routinely employed in our laboratories in the production of multimillicurie amounts of 11C labeled D,L amino acids in more than 90 production runs, with typical final D,L-product activities of 240-400 mCi. The high pressure system, however, is not restricted only to the production of 11C labeled amino acids, but for application in any synthetic procedure requiring high pressure (approximately 300 psia) and temperature (greater than 300 degrees C). PMID- 3027003 TI - Synthesis of DL-[1-11C]methionine. PMID- 3027004 TI - Pathophysiology of heart failure. PMID- 3027005 TI - Can 'mild' periodate oxidation be used for the specific histochemical identification of sialic acid residues? PMID- 3027006 TI - Dietary fibre intakes in the United Kingdom before and after retirement from work. AB - During a 4-year longitudinal nutritional survey at the age of retirement from work, 94 subjects participated, both before and after retirement. Investigations included 7-day weighed dietary records, questionnaire interviews and health screening. A further 89 subjects added to the data by questionnaire interviews. Dietary fibre intakes calculated from the 7-day weighed dietary records were: pre retirement 17.6 +/- 6.5 g/day, range 7-35 g; post-retirement 18.4 +/- 6.1 g/day, range 7-34 g. Before their retirement 88 per cent of the sample were not reaching the NACNE short-term recommendation of 25 g/day and 95 per cent had not reached the long-term recommendation of 30 g/day. There was little change in these percentages after retirement from work. Those individuals whose intakes were greater than 30 g/day had sometimes adopted a somewhat unusual style of eating. The percentage contributions to dietary fibre intake from the main food groups remained consistent, with vegetables and breads as the major sources, followed in importance by breakfast cereals and fruits. In spite of the ready availability of higher fibre foods, and publicity from the mass media and nutritional counselling, the increased awareness of the role of dietary fibre in the prevention of constipation had not, for the majority, altered food choice. There was a significantly higher intake of dietary fibre when breakfast was eaten daily but no significant effect on intake with alterations in food choice caused by dentures, chewing difficulties, indigestion or weight control.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3027007 TI - A dietary survey of an isolated population in the UK: the islanders of Orkney. AB - Isolated communities, such as those in the Highlands and Islands of Scotland have a dietary pattern and lifestyle which may differ from the rest of the country. We have studied the diets of 118 Orkney Islanders (78 women, 40 men) over 2 non consecutive weeks, using a semi-weighed methodology. 'High tea', soups, fish, potatoes and bakery goods were features of the traditional dietary pattern. Fresh fruit and vegetable tended to be low, and intakes of vitamin C and, to a lesser extent, dietary fibre, were low in relation to energy. Energy intakes of men were consistent with an active lifestyle, while fat provided 42 per cent of the energy from food. The implications of these findings are discussed in relation to the COMA recommendations and the prevalence of heart disease in this region of Britain. PMID- 3027009 TI - Antimutagenic properties of WR 2721 and of a radioprotective mixture, ATP-AET serotonin, with regard to X ray induced reciprocal translocations in mouse spermatogonia. AB - Pretreatment by intraperitoneal administration of WR 2721 at 400 mg/kg body weight in mice receiving 4.0 Gy X rays was found to have an appreciable antimutagenic effect with regard to reciprocal translocation induction in spermatogonia. The effectiveness of the product tested proved superior to that of a radioprotective mixture of ATP-AET-serotonin given at optimal dose ratio--360, 24, and 8 mg/kg body weight, respectively. The RF (Reduction Factor) was 2.4 for WR 2721 and 1.8 for the mixture. The effect observed indicated WR 2721 to have potential capabilities for reducing the genetic risk of radiation in male individuals. PMID- 3027010 TI - Public health concerns from human exposure to oncogenic avian herpesviruses. PMID- 3027008 TI - [Recurrence tendency and malignant transformation of pleomorphic adenomas]. AB - Young patients and those with the stroma-rich variant of pleomorphic adenoma have an increased risk of local recurrence. Review of 98 patients with recurrent pleomorphic adenoma showed that the primary operation is nevertheless decisive for the further progress of the disease: incomplete tumour excision and enucleation of pleomorphic adenomas were responsible for tumour recurrence which may be multiple. Recurrence in turn favours the development of carcinoma within pleomorphic adenomas. Of 23 carcinomas, 20 developed within recurrent tumours. These 20 tumours comprised 16.9% of all recurrences observed. A mean time of 16 years relapsed after the primary operation before a carcinoma developed in the recurrence. Slow and protracted tumour growth were characteristic of those carcinomas that could be also demonstrated in the original tumour (0.9% of the group of non-recurrent tumours) with a mean time of 7 years before operation. Immediate tumour removal is therefore the best prophylaxis against carcinoma. In 19 patients we showed that local recurrence due to dissemination can be avoided by appropriate measures, even after intra-operative opening of the tumour capsule. PMID- 3027011 TI - DC potentials in different cells of the stria vascularis measured in vitro. AB - Fine glass microelectrodes were inserted into the stria vascularis (SV) from the endolymphatic side under conditions in vitro. DC potentials were recorded in individual cells of the SV. The electrophoretic dye marking technique was used for identification of cells with different DC potentials. Penetration of the luminal membrane of marginal cells (MC) was accompanied by an unstable transient negative potential of about -15 mV. When the electrode penetrated further into the MC, a positive potential of about +10 mV was recorded. In the basal cells (BC) of the SV a negative DC potential of approx. -45 mV was always measured. Addition of ouabain into the perfusion solution (10(-3) mol/l) abolished the positive potential of MC, whereas the negative potential of BC decreased to approximately one half of the original value. A higher positive potential (+17 mV) was found in the MC of animals which were exposed to noise twenty days before the recording. Results are discussed in the light of the knowledge about localization of different transporting systems within the SV cells. PMID- 3027012 TI - Polymyxin B and polymyxin B nonapeptide alter cytoplasmic membrane permeability in Escherichia coli. AB - The effects of polymyxin B and polymyxin B nonapeptide (PMBN) on the permeability of the Escherichia coli cytoplasmic membrane were investigated. Both compounds caused loss of free amino acids, uracil and K+ from E. coli. The rates of loss promoted by polymyxin B were one and a half to two-fold greater than those caused by PMBN. Although PMBN mediated loss of low molecular weight substances from E. coli, it was not bactericidal. In contrast, polymyxin B treated E. coli lysed and rapidly lost viability. We suggest that the bactericidal activity of polymyxin B may be related to its previously reported ability to release cytoplasmic proteins from bacteria. PMID- 3027013 TI - Evolution of R plasmids by replicon fusion. AB - The occurrence of plasmid-plasmid fusions in nature and their possible influence on the spread of bacterial drug resistance is considered. Three ways in which replicon fusions can be formed are described; specifically fusions mediated by IS elements, "one-ended transposition" systems derived from Tn3 and like transposons, and pUB2380, a prototype for a new type of transposition. Non transposition based systems which mediate replicon fusion are also briefly considered. PMID- 3027014 TI - Antibiotic resistance profiles and molecular epidemiology of Salmonella typhimurium and S. dublin, mainly from cattle. AB - Among 130 strains of Salmonella typhimurium and 191 strains of S. dublin, all isolated from cattle in the early 1980s, 80% and 63%, respectively, were resistant to one or more (up to three) antibiotics. Monoresistance to sulphonamides was most common. In 169 strains of the two serotypes from 1985 the antibiotic resistance load was of a similar size. The latter batch was not studied with respect to plasmids. In the strains from 1980 and 1981 the plasmid load was analysed by conjugation, transformation, extraction of plasmid DNA and subsequent electrophoresis in agarose gels. Plasmid DNA from 38 strains was further analysed by restriction with endonuclease EcoRI. On the basis of this the strains were classified into four groups: two of S. typhimurium, one of S. dublin and one group of both serotypes. This suggested that dissemination of strains and plasmids mainly occurred through clonal spread of strains. PMID- 3027015 TI - Molecular biology of transposable elements. AB - Transposable elements play an essential part in the epidemic spread of antibiotic resistances among bacterial populations. Their ubiquity and their role in the molecular evolution of genomes and organisms has captured the interest of molecular biologists, who investigate the genetic structure and novel mechanisms of recombination underlying transposition. This article, after introducing the general structure of mobile genetic elements and basic mechanistic concepts of transposition in bacteria, uses the well-studied Tn3 family as paradigm for gene organization, gene function and gene regulation. Finally, new applications of transposons in genetic engineering will be described. PMID- 3027016 TI - Role of transposition and homologous recombination in the rearrangement of plasmid DNA. AB - The multi-resistance plasmid pBP16 was used to analyse the variability of R factors from clinical isolates and the molecular structures and processes involved. The observed rearrangement in pBP16 and its derivatives included inversion, deletion, replicon fusion and dissociation, and transposition. All these events could be traced to the presence and activity of multiple copies of the IS-element IS160 within pBP16. Since IS-elements are common in R-factors, it is likely that they are one of the main reasons for plasmid instability and that they are involved in a major way in plasmid evolution. PMID- 3027017 TI - Plasmid-mediated resistance to beta-lactam antibiotics in gram-negative bacteria: the role of in-vivo recyclization reactions in plasmid evolution. AB - Over 20 different plasmid-encoded beta-lactamases have so far been discovered. This paper considers genetic mechanisms by which beta-lactamase genes encoded by plasmids are disseminated across generic boundaries. Particular emphasis is placed on the evolution of plasmids carrying all or part of Tn3 and encoding TEM 1 beta-lactamase in Haemophilus and Neisseria species. Examples of the acquisition of broad host range plasmids carrying Tn3 sequences and of rescue of transposon sequences to indigenous plasmids or to the host chromosome have been found in these two genera. Studies on nonconjugative beta-lactamase plasmids in Neisseria and Haemophilus are consistent with the evolution of a family of plasmids originating from the insertion of Tn3 into an indigenous progenitor plasmid. A series of subsequent insertional and deletional events, most probably occurring as a consequence of genetic transfer, have given rise to the existing group of small, closely related ampicillin-resistance plasmids found currently in these genera. A general model for deletional and other rearrangements caused by recombinational recyclization during the evolution of resistance plasmids is described. PMID- 3027019 TI - Insertions of resistance genes into Tn21-like transposons. AB - The evolution of Tn21 and related multiresistance transposons of natural and artificial origin is described. Variations resulting from insertions and deletions in these transposons indicate the presence of specific recombination sites. There is evidence that these 'hot spots' are flanking several resistance genes. We examined these structures on the basis of the aadA gene, mediating resistance to spectinomycin and streptomycin. Comparison of the DNA sequences of Tn21, Tn7 and pSa indicate the wide spread of these recombination sites. PMID- 3027018 TI - Evolution of transposable elements. AB - Transposable elements are at an advantage compared with other DNA sequences. This has led to the generation of transposable systems from a variety of different sources. Once fabricated, these elements can very simply acquire accessory genes, such as those that encode resistance to antibiotics. This can lead to further evolutionary advantages and to their shorter-term importance. It is likely that many new transposable systems will be discovered, owing to the advantages conferred on sequences by transposition, and it is certain that many new transposons that encode varying drug resistance patterns will arise. PMID- 3027020 TI - Evolution and transfer of aminoglycoside resistance genes under natural conditions. AB - 3'-Aminoglycoside phosphotransferases [APH(3')] were chosen as a model to study the evolution and the transfer of aminoglycoside resistance genes under natural conditions. Comparison of the amino acid sequences of APH(3') enzymes from transposons Tn903 (type I) and Tn5 (type II) detected in Gram-negative bacteria, from the Gram-positive Staphylococcus and Streptococcus (type III), from the butirosin-producing Bacillus circulans (type IV) and from a neomycin-producing Streptomyces fradiae (type V) indicate that they have diverged from a common ancestor. These structural data support the hypothesis that the antibiotic producing strains were the source of certain resistance determinants. We have shown that kanamycin resistance in Campylobacter coli BM2509 was due to the synthesis of an APH(3')-III, an enzyme not detected previously in a Gram-negative bacterium. The genes encoding APH(3')-III in Streptococcus and Campylobacter are identical. These findings constitute evidence for a recent in-vivo transfer of DNA between Gram-positive and Gram-negative bacteria. PMID- 3027021 TI - Firing properties and hypercapnic responses of single phrenic motor axons in the rat. AB - Inspiratory phase activity was recorded from 33 phrenic motoneuron (PM) axonal fibers in anesthetized, vagotomized, artificially ventilated adult rats. During control conditions (no inspired CO2 added), the population of PM fibers could be separated into early and late onset types based on the time of firing onset relative to the onset of whole phrenic nerve activity. Mean discharge frequencies of both types were not significantly different. Compared with late PM's, early PM's had more spikes per inspiration, fired for a longer period, and the last spike occurred later and during the postinspiratory period. Further, the mean minimal interspike interval was shorter for early PM's than for late PM's. Increasing inspired CO2 to 0.03 and 0.05 resulted in earlier firing onsets and a greater number of spikes per inspiration, particularly for late PM's. Increases in mean firing frequency occurred for both PM types. Mean minimal interspike intervals for both types of PM's showed progressive reductions as CO2 rose. For almost all of the firing properties examined in this study, responses of rat PM axons were similar to those previously reported for the cat. PMID- 3027022 TI - Use of the doubly labeled water technique in humans during heavy sustained exercise. AB - We measured energy expenditure with the doubly labeled water technique during heavy sustained exercise in the Tour de France, a bicycle race lasting more than 3 wk. Four subjects were observed for consecutive intervals of 7, 8, and 7 days. Each interval started with an oral isotope dose to reach an excess isotope level of 200 ppm 18O and 130 ppm 2H. The biological half-lives of the isotopes were between 2.25 and 3.80 days. Energy expenditure was compared with simultaneous measurements of energy intake, and body mass and body composition did not change significantly. The doubly labeled water technique gave higher values for energy expenditure than the food record technique. The discrepancy showed a systematic increment from the first to the third interval, being 12.9 +/- 7.9, 21.4 +/- 9.8, and 35.3 +/- 4.4% of the energy expenditure calculated from dietary intake, respectively. Possible explanations for the discrepancy are discussed. The subjects reached an average daily metabolic rate of 3.4-3.9 or 4.3-5.3 times basal metabolic rate based on the food record technique and the doubly labeled water technique, respectively. Thus, when measured with the same technique, the energetic ceiling for performance in humans is comparable with that of animals like birds. PMID- 3027023 TI - Effect of complement activation with cobra venom factor on pulmonary vascular permeability. AB - We examined the effect of acute complement activation on lung vascular permeability to proteins in awake sheep prepared with lung lymph fistulas. Complement was activated by cobra venom factor (CVF) infusion (400 U/kg for 1 h iv). Studies were made in two groups of sheep: 1) infusion of CVF containing the endogenous phospholipase A2 (PLA2) (n = 6); and 2) infusion of CVF pretreated with bromophenacyl bromide to inhibit PLA2 activity (n = 5). Intravascular complement activation transiently increased mean pulmonary arterial pressure (Ppa) and pulmonary vascular resistance (PVR) in both groups. Pulmonary lymph flow (Qlym) and lymph protein clearance (Qlym X lymph-to-plasma protein concentration ratio) were also transiently increased in both groups. Pulmonary vascular permeability to proteins was assessed by raising left atrial pressure and determining the lymph-to-plasma protein concentration ratio (L/P) at maximal Qlym. In both groups the L/P at maximal Qlym was not different from normal. In a separate group (n = 4), CVF-induced complement activation was associated with 111In-oxine granulocyte sequestration in the lungs. In vitro plasma from CVF treated animals aggregated neutrophils but did not stimulate neutrophils to produce superoxide anion generation. Therefore, CVF-induced complement activation results in pulmonary neutrophil sequestration and in increases in PVR and lymph protein clearance. The increase in lymph protein clearance is due to increased pulmonary microvascular pressure and not increased vascular permeability to proteins. PMID- 3027026 TI - [Arterial computerized tomography of the liver compared to conventional computerized tomography and arteriography]. PMID- 3027024 TI - Hormone-responsive cells derived from human dental papilla: characterization in vitro and in vivo in diffusion chambers. AB - Cells of the dental papilla are capable of odontoblastic, fibroblastic, and endothelial differentiation and formation of dentin and the dental pulp. In the present study dental papilla cells, obtained from human tooth buds (HDP cells), were cultured in vitro through 3 to 7 passages. After exposure to prostaglandin E2 there was a marked decrease in intracellular cyclic AMP (cAMP) levels as compared to hormone-free controls. Parathyroid hormone and calcitonin had stimulatory effects with 1 and 2 log increases in cAMP, respectively. The HDP cells showed moderate activity of alkaline phosphatase, 1 log higher than that of hamster kidney fibroblasts (BHK 13) and 1 log lower than that of osteoblastic osteosarcoma cells (ROS 17/2). When cultured for 4 or 8 wk in diffusion chambers (DC) implanted in athymic mice, many of the HDP cells underwent odontoblastic morphodifferentiation with very long, single processes extending into the matrix. This matrix contained banded and unbanded collagen fibers. Neither light nor electron microscopy of the DC content revealed mineral deposits. These results suggest that HDP cells have an intrinsic potential for partial odontoblastic differentiation; inductive signals like those originating from odontogenic epithelium are probably essential for the completion of hard tissue formation. PMID- 3027027 TI - [Computerized portography in the evaluation of malignant hepatic tumors]. PMID- 3027025 TI - bFGF is the putative natural growth factor for human melanocytes. AB - Normal human melanocytes, unlike pigment cells from metastatic melanomas, do not survive in culture in routine, serum-supplemented media. The search for natural growth factors for melanocytes has shown that mitogenic activity is ubiquitous in several tissues and in melanomas. Of several known growth factors tested, basic fibroblast growth factor (bFGF) was the only one mitogenic for melanocytes but only in the presence of cyclic-adenosine-monophosphate (cAMP) stimulators. The mitogenic activity toward melanocytes in tissues and melanoma cell extracts had high affinity for heparin and antibodies to bFGF synthetic peptides. These results suggest that one of the growth factors for melanocytes might be bFGF or a bFGF-like polypeptide and that autocrine production of bFGF-like molecules by melanoma cells may contribute to the malignant phenotype of melanocytes. Because acidic FGF (aFGF) did not stimulate growth, the receptors for bFGF on melanocytes might be significantly different from those for a FGF. PMID- 3027028 TI - [Glucagonoma and an associated dermatological syndrome]. PMID- 3027030 TI - Molecular cloning of copper resistance genes from Pseudomonas syringae pv. tomato. AB - A cosmid library of copper-resistant (Cur) Pseudomonas syringae pv. tomato PT23 plasmid DNA was constructed and mobilized into the copper-sensitive recipient P. syringae pv. syringae PS61. One resultant cosmid clone, pCOP1 (46 kilobases), conferred copper resistance. The PT23 Cur gene(s) was located on pCOP1 by subcloning PstI restriction endonuclease fragments of pCOP1 in the broad-host range vector pRK404. A subclone containing a 4.4-kilobase PstI fragment conferred Cur on PS61. The Cur gene(s) was further located by insertional inactivation with Tn5. A subcloned fragment internal to the Cur determinant on pCOP2 was probed to plasmid and chromosomal DNA of four copper-resistant and three copper-sensitive strains of P. syringae pv. tomato. The probe hybridized to plasmids in resistant strains, but showed no detectable homology to copper-sensitive strains. PMID- 3027029 TI - Effects of plasmid propagation of a sporulation promoter on promoter utilization and sporulation in Bacillus subtilis. AB - Transcription of the sporulation gene spoVG of Bacillus subtilis is induced at the onset of spore formation and depends on the products of the regulatory genes spoOA, spoOB, and spoOH. We describe two effects of propagating the promoter region of spoVG on a multicopy plasmid replicon in B. subtilis cells. One effect is that transcription from the plasmid-borne spoVG promoter is altered with respect to the time of its induction and the dependence on spoO gene products. An example of this effect is that plasmid propagation was observed to relieve substantially the inhibitory effect of a mutation in spoOH, the spoO gene upon which spoVG promoter activity is most strongly dependent. We present results which suggest that propagation on a plasmid replicon causes an alteration in the conformation of spoVG promoter DNA which somehow compensates for the defective spoOH gene product. Plasmid propagation did not, however, entirely eliminate the requirement for the spoOH gene product; little or no spoVG-directed RNA synthesis was observed in cells bearing a putative spoOH deletion mutation, a finding which indicates that SpoOH protein plays an indispensable role in spoVG promoter utilization. Another effect of propagating the promoter region of spoVG on a multicopy plasmid is to inhibit sporulation. S1 nuclease mapping experiments suggest that amplification of spoVG on a multicopy plasmid causes the titration of a transcription factor or minor form of RNA polymerase holoenzyme required for utilization of one of the two overlapping promoters which comprise the spoVG transcription initiation region. PMID- 3027031 TI - Cloning and characterization of the aerobic sn-glycerol-3-phosphate dehydrogenase structural gene glpD of Escherichia coli K-12. AB - The glpD gene encoding aerobic sn-glycerol-3-phosphate dehydrogenase of Escherichia coli K-12 was cloned into pACYC177 from a lambda glpD transducing phage. The recombinant plasmid, designated pSH55, carried a 7.4-kilobase-pair HindIII fragment containing the glpD and glpR genes. The glpD gene was subcloned into pACYC177 on a 4.4-kilobase-pair BamHI-HindIII fragment. Expression of the cloned glpD gene was regulated in the manner previously described for the chromosomal glpD gene. The position of glpD on this plasmid was determined by Tn1000 insertional inactivation experiments. The glpD gene product, a polypeptide of Mr 55,000, was detected in a maxicell system. Truncated polypeptides replaced the 55,000-molecular-weight polypeptide when plasmid derivatives harboring Tn1000 insertions that inactivate glpD were used as templates. The sizes of these polypeptides confirmed the previously determined direction of transcription and allowed estimation of the translation start site. Determination of the apparent Mr of a hybrid protein encoded by a glpD'-'lacZ fusion provided additional evidence for the position of the glpD control region. The amino-terminal 30 to 60 amino acids of this hybrid protein (provided by glpD) were sufficient for efficient membrane localization of glpD'-'lacZ-encoded beta-galactosidase activity. The glpD3 mutation was mapped within the glpD gene, providing additional evidence that glpD is the structural gene for aerobic sn-glycerol-3 phosphate dehydrogenase. PMID- 3027032 TI - Divergent transcription of the sn-glycerol-3-phosphate active transport (glpT) and anaerobic sn-glycerol-3-phosphate dehydrogenase (glpA glpC glpB) genes of Escherichia coli K-12. AB - The glpTQ operon and the glpA and glpB genes are located adjacent to one another near min 49 of the linkage map of Escherichia coli K-12. The positions and directions of transcription of the glpA and glpB genes with respect to the glpTQ operon were determined in the present work. Strains harboring Mu d1(Ap lac) fusions in either glpA or glpB were converted to the respective lambda p1(209) lysogens. Induction of these lysogens with mitomycin C resulted in production of Lac+ phage progeny which carried adjacent chromosomal DNA. Genetic crosses with a collection of glpT mutant strains were performed with several such phage lines. A fine-structure deletion map of the glpT gene was thus constructed. All phages used for this mapping carried DNA starting with the promoter-proximal end of glpT. This indicated that the glpTQ operon and the glpA and glpB genes are transcribed divergently. Additional evidence supporting this conclusion was obtained by physical mapping of restriction endonuclease cleavage sites in plasmids carrying these genes and in plasmids carrying glpA-lacZ or glpB-lacZ fusions. A new designation (glpC) for the gene encoding the 41,000-Mr subunit of the anaerobic sn-glycerol-3-phosphate dehydrogenase was proposed to distinguish it from the glpA gene, which encodes the 62,000-Mr subunit of the dehydrogenase, and the glpB gene, which encodes a membrane anchor subunit of the dehydrogenase. These three genes were present in an operon transcribed in the order glpA glpC glpB in the clockwise direction on the linkage map of E. coli. PMID- 3027033 TI - Effect of growth phase on phospholipid biosynthesis in Saccharomyces cerevisiae. AB - The effect of growth phase on the membrane-associated phospholipid biosynthetic enzymes CDP-diacylglycerol synthase, phosphatidylserine synthase, phosphatidylinositol synthase, and the phospholipid N-methyltransferases in wild type Saccharomyces cerevisiae was examined. Maximum activities were found in the exponential phase of cells grown in complete synthetic medium. As cells entered the stationary phase of growth, the activities of the CDP-diacylglycerol synthase, phosphatidylserine synthase, and the phospholipid N-methyltransferases decreased 2.5- to 5-fold. The subunit levels of phosphatidylserine synthase and the cytoplasmic-associated enzyme inositol-1-phosphate synthase were not significantly affected by the growth phase. When grown in medium supplemented with inositol-choline, cells in the exponential phase of growth had reduced CDP diacylglycerol synthase, phosphatidylserine synthase, and phospholipid N methyltransferase activities, with repressed subunit levels of phosphatidylserine synthase and inositol-1-phosphate synthase compared with cells grown without inositol-choline. Enzyme activity levels remained reduced in the stationary phase of growth of cells supplemented with inositol-choline. The phosphatidylserine synthase and inositol-1-phosphate synthase subunit levels, however, were depressed. Phosphatidylinositol synthase (activity and subunit) was not affected by growth in medium supplemented with or without inositol-choline or the growth phase of the culture. The phospholipid composition of cells in the exponential and stationary phase of growth was also examined. The phosphatidylinositol to phosphatidylserine ratio doubled in stationary-phase cells. The phosphatidylcholine to phosphatidylethanolamine ratio was not significantly affected by the growth phase of cells. PMID- 3027034 TI - Expression of a tRNA gene in the context of the lacZ mRNA. AB - Fusions of the gene for tyrosine suppressor tRNA, tyrT(Sup3), and the lacZ gene of Escherichia coli were constructed such that the tRNA gene could be expressed from either its own promoter or that of the lac operon. These chimeras, carried on phage M13 vectors, were tested for the expression of the tRNA in E. coli. The tRNA gene was expressed on the order of 10-fold more weakly from the lac promoter than from its own promoter. To examine whether pausing or premature termination of transcription played a role in determining the relative strength, the fusions were tested in a variety of genetic backgrounds and under different physiological conditions that uncouple transcription and translation. The expression of the tRNA was not enhanced in backgrounds in which polarity was weakened or under the other conditions tested, although a dependence on nusB function was observed when the tRNA was transcribed from the lac promoter. These results indicate that pausing or premature termination of transcription did not play a role in the weak expression of the gene fusions. The results further suggest that the transcription of the tyrT gene does not normally require relief from polarity as imposed by any of the known transcriptional termination systems, in contrast to the antitermination system thought to be involved in the expression of the rRNAs. PMID- 3027035 TI - Molecular characterization and nucleic acid sequence of an avirulence gene from race 6 of Pseudomonas syringae pv. glycinea. AB - A gene was previously cloned from Pseudomonas syringae pv. glycinea race 6, designated avirulence gene A (avrA), that controls the expression of virulence by the pathogen on specific cultivars of soybean. A 3.2-kilobase (kb) AccI subclone from the cosmid clone pPg6L3 was shown to be active when cloned into the broad host-range vector pRK404. Transposon Tn5 mutagenesis and deletion analysis delineated a span of approximately 2.5 kb of DNA that was necessary for gene activity. The nucleotide sequence of a 3.409-kb segment of DNA which contained the avrA gene has been determined. An open reading frame of 2.721 kb of DNA, which correlates with the region of DNA defined by transposon mutagenesis and deletion analysis, was identified. The open reading frame would encode a protein of 100.866 kilodaltons, which is in good agreement with the 100-kilodalton protein expressed by Escherichia coli maxicells. PMID- 3027036 TI - Cloning and sequencing of the blaZ gene encoding beta-lactamase III, a lipoprotein of Bacillus cereus 569/H. AB - It has not been clear whether the membrane-bound beta-lactamase III of Bacillus cereus 569 is a separate enzyme or a modified form of the secreted beta-lactamase I. The membrane enzyme is an acyl-glyceride thioether-linked lipoprotein (J. B. K. Nielsen and J. O. Lampen, Biochemistry 22:4652-4656, 1983) and thus is probably a separate entity. We cloned the beta-lactamase III gene (blaZ) on a 4.9 kilobase-pair ClaI fragment from mutant strain 569/H (constitutive for high-level production of beta-lactamases I, II, and III), and the nucleotide sequence was determined. The structural gene was flanked by typical promoter, transcription termination, and translation initiation sequences. Expression of the cloned gene in Escherichia coli was low in exponential-phase cultures and increased only as the cultures reached the stationary phase. The deduced amino acid sequence indicates a pre-beta-lactamase III of 316 amino acid residues (35,021 daltons), with a 29-residue signal peptide and a mature lipoprotein form of approximately 32,500 daltons. The 12 NH2-terminal residues of a 21-kilodalton tryptic peptide from the B. cereus membrane enzyme were in agreement with the sequence deduced from the cloned gene. The amino acid sequence was highly homologous to the class A beta-lactamases, especially that of Bacillus licheniformis 749. beta-Lactamase III is a distinct class A enzyme and the product of a separate gene (blaZ). PMID- 3027037 TI - Indirect role of adenylate cyclase and cyclic AMP in chemotaxis to phosphotransferase system carbohydrates in Escherichia coli K-12. AB - Most strains of Escherichia coli K-12 lacking the enzyme adenylate cyclase showed normal chemotaxis toward carbohydrates taken up and phosphorylated by the phosphoenolpyruvate-dependent carbohydrate: phosphotransferase system. The normal reaction was observed even in the absence of externally added cyclic adenosine 3',5'-phosphate, provided that the enzyme II chemoreceptors and the flagella were synthesized. In the CA8306 series of strains, however, the cya-854 deletion abolished chemotaxis toward phosphotransferase system carbohydrates even though growth on and transport of these carbohydrates were not affected. This abnormal phenotype was due to the presence of a specific mutation in strain CA8306 which mapped in or close to the crp locus and apparently prevented expression of a hitherto unidentified molecule involved in enzyme II-mediated signal transduction. This molecule is neither a pts protein nor a cyclic adenosine 3',5' phosphate-binding protein. PMID- 3027038 TI - Effects of deletion and insertion mutations in the ilvM gene of Escherichia coli. AB - A plasmid was constructed that carried the ilvG and ilvM genes and the associated promoter and leader regions derived from the K-12 strain of Escherichia coli. The ilvG gene contained a + 1 frameshift mutation that enabled the plasmid to specify acetohydroxyacid synthase II. The plasmid was modified by deletions in the terminus of and within the ilvM gene and by insertions into the ilvM gene. The effects of these modifications on the phenotypes of the plasmids were examined in a host strain that lacked all three isozymes of acetohydroxyacid synthase. Most of the ilvM mutant plasmids so obtained permitted growth of the host strain in the absence of isoleucine but not in the absence of valine. Growth in the presence of valine, however, was very slow. No significant acetohydroxyacid synthase activity could be detected even when the cells were grown in a valine supplemented minimal medium. It thus appears that, at most, only a very low level of acetohydroxyacid synthase activity occurred with ilvG in the absence of ilvM and that low activity was more effective for acetohydroxy butyrate formation than for acetolactate formation. The ilvM gene product could be formed under the control of the lac promoter in the presence of a plasmid that carried an in-frame gene fusion between lacZ and the downstream portion of ilvG. Extracts from the host strain that contained such an IlvG(-)-IlvM+ plasmid could be combined with extracts from cells that contained one of the IlvG+-IlvM- plasmids to yield acetohydroxyacid synthase activity. Thus, the ilvM and ilvG genes could be expressed independently of each other. PMID- 3027039 TI - Sequence of the Ampullariella sp. strain 3876 gene coding for xylose isomerase. AB - The nucleotide sequence of the gene coding for xylose isomerase from Ampullariella sp. strain 3876, a gram-positive bacterium, has been determined. A clone of a fragment of strain 3876 DNA coding for a xylose isomerase activity was identified by its ability to complement a xylose isomerase-defective Escherichia coli strain. One such complementation positive fragment, 2,922 nucleotides in length, was sequenced in its entirety. There are two open reading frames 1,182 and 1,242 nucleotides in length, on opposite strands of this fragment, each of which could code for a protein the expected size of xylose isomerase. The 1,182 nucleotide open reading frame was identified as the coding sequence for the protein from the sequence analysis of the amino-terminal region and selected internal peptides. The gene initiates with GTG and has a high guanine and cytosine content (70%) and an exceptionally strong preference (97%) for guanine or cytosine in the third position of the codons. The gene codes for a 43,210 dalton polypeptide composed of 393 amino acids. The xylose isomerase from Ampullariella sp. strain 3876 is similar in size to other bacterial xylose isomerases and has limited amino acid sequence homology to the available sequences from E. coli, Bacillus subtilis, and Streptomyces violaceus-ruber. In all cases yet studied, the bacterial gene for xylulose kinase is downstream from the gene for xylose isomerase. We present evidence suggesting that in Ampullariella sp. strain 3876 these genes are similarly arranged. PMID- 3027040 TI - Repressor gene finO in plasmids R100 and F: constitutive transfer of plasmid F is caused by insertion of IS3 into F finO. AB - Fertility factor F confers bacterial conjugation, a process which involves at least 20 tra genes. Resistance plasmids such as R100, R6-5, and R1 have homology with F in the tra region. Conjugal transfer of these plasmids is, however, repressed, while transfer of F is constitutive. Repression of R transfer is due to the existence of the two genes, called finO and finP; constitutive transfer of F is believed to be due to a lack of finO in F. In this paper, we report the identification and DNA sequence of the finO gene of R100, encoding a protein of 21,265 daltons. We show that F does actually encode finO, but the gene has been inactivated by insertion of IS3. Lederberg and Tatum (Nature [London] 158:558, 1946), who discovered sexuality in bacteria, may have had an Escherichia coli K 12 strain harboring such an finO F factor, which facilitated the generation of recombinant progeny useful for genetic analysis of bacteria and established the foundation for molecular genetics. PMID- 3027041 TI - Tn2501, a component of the lactose transposon Tn951, is an example of a new category of class II transposable elements. AB - Tn2501 is a cryptic class II transposon found as part of the lactose transposon Tn951. Insertional inactivation and nucleotide sequence analysis of Tn2501 allowed us (i) to localize the transposase (tnpA) and the resolvase (tnpR) genes as well as the resolution site (res) of Tn2501 and (ii) to compare Tn2501 with other well-known elements of the two subgroups of class II transposons (Tn3, gamma delta, Tn951, IS101; and Tn21, Tn501, Tn1721). The genetic organization of Tn2501 is similar to that of Tn3 with divergent transcription of the tnpA and tnpR genes away from the intervening res site. The tnpR gene of Tn2501 shows weak homology with that of Tn3 and even less with those of Tn21 and Tn501. However, the tnpA gene and the inverted repeat sequences of Tn2501 present more homology with those of Tn21 and Tn501 than with those of Tn3. Complementation studies showed that TnpA- mutants of Tn2501 can be complemented, at a low frequency, by the Tn21 transposase. None of the tested transposons complemented TnpR- mutants of Tn2501. PMID- 3027042 TI - Generation of deletions in the 3'-flanking sequences of the Escherichia coli crp gene that induce cyclic AMP suppressor functions. AB - The crp structural gene and its 3'-flanking sequences were subcloned into M13mp8, and in vitro deletions were constructed in both the 5' and 3' ends of the gene by using Bal 31 nuclease. Deletions ranged in size from 24 to 250 base pairs at the 5' end of crp. Sixteen deletions generated at the 3' end of the gene ranged in size from 133 to 675 base pairs. The majority of deletions extended into the crp structural gene. Another class of deletions, i.e., delta crp-4, delta crp-17, and delta crp-2, had endpoints extending in the 3'-flanking sequences external to the crp structural gene. Deletions were subcloned into pBR322 and transformed into the Escherichia coli cya crp deletion strain NCR438. Transformants containing plasmid pBM4 with the delta crp4 mutation, a deletion of 133 base pairs, were cyclic AMP independent. Strain NCR440 harboring this plasmid expressed beta galactosidase and threonine dehydratase activities and fermented lactose, ribose, arabinose, and xylose in the absence of exogenous cyclic AMP. The delta crp-4 mutation also caused strain NCR440 to be hypersensitive to exogenous cyclic AMP. The cylic AMP receptor protein expressed in maxicells from pBM4 carrying the delta crp-4 mutation comigrated with the wild-type protein on electrophoretic gels. The delta crp-4 mutation demonstrates that sequences distal to the crp structural gene can mediate cyclic AMP suppressor functions. PMID- 3027043 TI - Characterization of bromoethanesulfonate-resistant mutants of Methanococcus voltae: evidence of a coenzyme M transport system. AB - Mutants of Methanococcus voltae were isolated that were resistant to the coenzyme M (CoM; 2-mercaptoethanesulfonic acid) analog 2-bromoethanesulfonic acid (BES). The mutants displayed a reduced ability to accumulate [35S]BES relative to the sensitive parental strain. BES inhibited methane production from CH3-S-CoM in cell extracts prepared from wild-type sensitive or resistant strains. BES uptake required the presence of both CO2 and H2 and was inhibited by N-ethylmaleimide and several reagents that are known to disrupt energy metabolism. The mutants showed normal uptake of isoleucine and were not cross-resistant to either azaserine or 5-methyltryptophan and, thus, were neither defective in general energy-dependent substrate transport nor envelope permeability. Both HS-CoM and CH3-S-CoM prevented the uptake of BES and protected cells from inhibition by it. We propose that M. voltae has an energy-dependent, carrier-mediated uptake system for HS-CoM and CH3-S-CoM which can also mediate uptake of BES. PMID- 3027044 TI - DNA sequence and in vitro expression of the B875 light-harvesting polypeptides of Rhodobacter sphaeroides. AB - The genes for the Rhodobacter sphaeroides light-harvesting B875-beta, and B875 alpha polypeptides (pufB and pufA) are closely linked to the genes for the reaction center L and reaction center M polypeptides (pufL and pufM) on what has been termed the puf operon (gene order, pufB, A, L, M). The DNA sequence of the pufB and pufA structural genes from wild-type R. sphaeroides 2.4.1 was determined and aligned with the available amino acid sequence of the wild-type B875-beta and B875-alpha polypeptides. The relative levels of the B875-beta and B875-alpha and the reaction center L and reaction center M polypeptides synthesized in a homologous cell-free transcription-translation system were compared with those found in vivo. Analysis of the gene products produced in vitro with plasmids containing deletions upstream of the pufB structural gene identified a region of DNA required for expression of the B875-beta and B875-alpha polypeptides. These results support the hypothesis that the mapped 5' termini of the large and small puf operon transcripts represent transcription initiation sites. PMID- 3027045 TI - Processing of the initiation methionine from proteins: properties of the Escherichia coli methionine aminopeptidase and its gene structure. AB - Methionine aminopeptidase (MAP) catalyzes the removal of amino-terminal methionine from proteins. The Escherichia coli map gene encoding this enzyme was cloned; it consists of 264 codons and encodes a monomeric enzyme of 29,333 daltons. In vitro analyses with purified enzyme indicated that MAP is a metallo oligopeptidase with absolute specificity for the amino-terminal methionine. The methionine residues from the amino-terminal end of the recombinant proteins interleukin-2 (Met-Ala-Pro-IL-2) and ricin A (Met-Ile-Phe-ricin A) could be removed either in vitro with purified MAP enzyme or in vivo in MAP-hyperproducing strains of E. coli. In vitro analyses of the substrate preference of the E. coli MAP indicated that the residues adjacent to the initiation methionine could significantly influence the methionine cleavage process. This conclusion is consistent, in general, with the deduced specificity of the enzyme based on the analysis of known amino-terminal sequences of intracellular proteins (S. Tsunasawa, J. W. Stewart, and F. Sherman, J. Biol. Chem. 260:5382-5391, 1985). PMID- 3027046 TI - Common mechanism of ampC beta-lactamase induction in enterobacteria: regulation of the cloned Enterobacter cloacae P99 beta-lactamase gene. AB - Expression of the chromosomal beta-lactamase from the ampC gene in inducible in both Enterobacter cloacae and Citrobacter freundii. Cloning of ampC as well as its regulatory gene, ampR, from E. cloacae P99 revealed a gene organization indentical to that of C. freundii in the corresponding region. Although almost no similarities could be found between the restriction maps of ampC and ampR in the two species, the genes cross-hybridize. Also, both ampR gene products have a size of about 31,000. The regulatory features of E. cloacae beta-lactamase induction are very similar to those in C. freundii, i.e., beta-lactamase synthesis is repressed by AmpR in the absence, and stimulated in the presence, of inducer. The AmpR function can be transcomplemented between the two species, but there are quantitative regulatory aberrations in such hybrids, in contrast to the total complementation obtained within each system. These results suggest that the mechanism of beta-lactamase induction is the same in E. cloacae, C. freundii, and other gram-negative bacteria with inducible chromosomal beta-lactamase expression. PMID- 3027047 TI - Gene organization of the first catabolic operon of TOL plasmid pWW53: production of indigo by the xylA gene product. AB - The entire operon coding for the enzymes responsible for conversion of toluenes to benzoates has been cloned from TOL plasmid pWW53 and the position of the genes accurately located. The coding region was 7.4 kilobase pairs (kbp) long, and the gene order was operator-promoter region (OP1)-a small open reading frame-xylC (1.6 kbp)-xylA (2.9 kbp)-xylB (1.8 kbp). Within the coding region there was considerable homology with the isofunctional region of the archetypal TOL plasmid pWW0. A central region of 2.9 kbp complemented an xylA (for xylene oxygenase) mutant of Pseudomonas putida mt-2 and was also capable of conferring the ability to convert indole to indigo on strains of Escherichia coli and P. putida. This reaction has been reported previously only for dioxygenases involved in aromatic catabolism but not for monooxygenases. It is proposed that the region encodes xylene oxygenase activity capable of direct monohydroxylation of indole to 3 hydroxyindole (oxindole), which then spontaneously dimerizes to form indigo. PMID- 3027049 TI - Lipid acyl chain-dependent effects of sterols in Acholeplasma laidlawii membranes. AB - Acholeplasma laidlawii was grown with different fatty acids for membrane lipid synthesis (saturated straight- and branched-chain acids and mono- and di unsaturated acids). The ability of 12 different sterols to affect cell growth, lipid head group composition, the order parameter of the acyl chains, and the phase equilibria of in vivo lipid mixtures was studied. The following two effects were observed with respect to cell growth: with a given acyl chain composition of the membrane lipids, growth was stimulated, unaffected, reduced, or completely inhibited (lysis), depending on the sterol structure; and the effect of a certain sterol depended on the acyl chain composition (most striking for epicoprostanol, cholest-4-en-3-one, and cholest-5-en-3-one, which stimulated growth with saturated acyl chains but caused lysis with unsaturated chains). The three lytic sterols were the only sterols that caused a marked decrease in the ratio between the major lipids monoglucosyldiglyceride and diglucosyldiglyceride and hence a decrease in bilayer stability when the membranes were enriched in saturated (palmitoyl) chains. With these chains correlations were found for several sterols between the glucolipid ratio and the order parameter of the acyl chains, as well as the lamellar-reversed hexagonal phase transition, in model systems. A shaft experiment revealed a marked decrease in the ratio of monoglucosyldiglyceride to diglucosyldiglyceride with the lytic sterols in unsaturated (oleoyl) membranes. The two cholestenes induced nonlamellar phases in in vivo mixtures of oleoyl A. laidlawii lipids. The order parameters of the oleoyl chains were almost unaffected by the sterols. Generally, the observed effects cannot be explained by an influence of the sterols on the gel-to-liquid crystalline phase transition. PMID- 3027048 TI - Gene encoding the 37,000-dalton minor sigma factor of Bacillus subtilis RNA polymerase: isolation, nucleotide sequence, chromosomal locus, and cryptic function. AB - We began an analysis of rpoF, the gene encoding the cryptic, 37,000-dalton minor sigma factor (sigma-37) of Bacillus subtilis RNA polymerase. Using antibody raised against sigma-37 holoenzyme to probe a lambda gt11 expression vector library, we isolated a 901-base-pair EcoRI fragment that expressed the COOH terminal half of sigma-37 fused to lacZ. We used this fragment as a hybridization probe to isolate the entire rpoF gene and additional flanking sequences. Identity of the cloned gene was confirmed by the size and immunological reaction of its product expressed in Escherichia coli and, after DNA sequencing, by the homology of its predicted product (264 residues; 30,143 daltons) with other sigma factors. The DNA sequence also suggested that rpoF may lie in a gene cluster. Upstream of rpoF was an open reading frame that would encode a protein of 17,992 daltons; this frame overlapped the rpoF-coding sequence by 41 base pairs. Immediately following rpoF was a reading frame that would encode a protein of at least 20,000 daltons; expression of this region may be translationally coupled to that of rpoF. By plasmid integration and PBS1 transduction, we found the chromosomal locus of rpoF linked to ddl and dal at 40 degrees on the B. subtilis map and near no known lesions affecting growth regulation or development. Further, an rpoF null mutation resulting from gene disruption had no effect on cell growth or sporulation in rich medium, suggesting that sigma-37 may partly control a regulon not directly involved in the sporulation process. PMID- 3027050 TI - Organization of rRNA genes in Mycobacterium bovis BCG. AB - The number of rRNA genes in Mycobacterium bovis BCG was examined by Southern hybridization of end-labeled 5S, 16S, and 23S rRNAs with BamHI, PstI, and SalI digests of M. bovis BCG DNA. Each RNA probe gave only one radioactive band with three kinds of DNA digest. These results suggest that M. bovis BCG chromosomes may carry only a minimum set of rRNA genes. Hybridization of randomly labeled rRNAs with BamHI, PstI, SalI, BglII, and PvuII digests of DNA from the same organism supported these conclusions. The 6.4-kilobase-pair SalI fragment containing the entire structural genes for both 16S and 23S rRNAs was cloned into pBR322. The cloned fragment was characterized by restriction endonuclease mapping, DNA-RNA hybridization analysis, and the R-loop technique. The results indicated that the fragments contained rRNA genes in the following order: 16S, 23S, and 5S rRNA genes. No tRNA gene was detected in the spacer region between the 16S and 23S rRNA genes, but one was found downstream of the 23S rRNA and 5S rRNA genes. PMID- 3027051 TI - Nucleotide sequence encoding the flavoprotein and iron-sulfur protein subunits of the Bacillus subtilis PY79 succinate dehydrogenase complex. AB - The nucleotide sequence of a 2.7-kilobase segment of DNA containing the sdhA and sdhB genes encoding the flavoprotein (Fp, sdhA) and iron-sulfur protein (Ip, sdhB) subunits of the succinate dehydrogenase of Bacillus subtilis was determined. This sequence extends the previously reported sequence encoding the cytochrome b558 subunit (sdhC) and completes the sequence of the sdh operon, sdhCAB. The predicted molecular weights for the Fp and Ip subunits, 65,186 (585 amino acids) and 28,285 (252 amino acids), agreed with the values determined independently for the labeled Fp and Ip antigens, although it appeared that the B. subtilis Fp was not functional after expression of the sdhA gene in Escherichia coli. Both subunits closely resembled the corresponding Fp and Ip subunits of the succinate dehydrogenase (SDH) and fumarate reductase of E. coli in size, composition, and amino acid sequence. The sequence homologies further indicated that the B. subtilis SDH subunits are equally related to the SDH and fumarate reductase subunits of E. coli but are less closely related than are the corresponding pairs of E. coli subunits. The regions of highest sequence conservation were identifiable as the catalytically significant flavin adenine dinucleotide-binding sites and cysteine clusters of the iron-sulfur centers. PMID- 3027052 TI - Reduced transposition in rho mutants of Escherichia coli K-12. AB - Substantially reduced frequencies of transposition for the transposons Tn5 and Tn9 and the insertion sequences IS1 and IS5 were observed in several rho mutants of Escherichia coli K-12 compared with those observed in their isogenic wild-type counterparts. The lower transposition frequencies could be due to decreased supercoiling of DNA, to altered expression of required genes, or to aberrant transcription of transposon or target DNA resulting from the lack of transcription termination at Rho-sensitive sites in rho mutants. PMID- 3027053 TI - Variations between the nucleotide sequences of Tn1, Tn2, and Tn3 and expression of beta-lactamase in Pseudomonas aeruginosa and Escherichia coli. AB - Using S1 nuclease assays, we located the sites of initiation of transcription of the beta-lactamase gene on Tn1 and Tn2. Transcription in Tn2, like that in Tn3, occurred from the P3 promoter, whereas transcription in Tn1 was initiated by two stronger and overlapping promoters, Pa and Pb. The nucleotide sequences of Tn1 and Tn2 were determined over a 1,195-base-pair segment constituting most of the sequences of the tnpR and bla genes and the intervening region. There were six base-pair differences between Tn1 and Tn3. One in the bla regulatory region accounted for the presence of the Pa and Pb promoters, and another in the bla structural gene is consistent with the isoelectric focusing difference found between the Tn1 and Tn3 enzymes. In contrast, there were 24 base-pair differences between Tn2 and Tn3, most of them clustered in one segment of the tnpR gene. PMID- 3027054 TI - The human multidrug resistance (mdr1) gene. cDNA cloning and transcription initiation. AB - Multidrug resistance in human KB carcinoma cells selected for resistance to colchicine, vinblastine, or adriamycin results from overexpression, and frequently amplification, of a specific gene (mdr1). Overlapping cDNA clones representing a complete 4.7-kilobase mdr1 transcript have been obtained from multidrug-resistant KB cells. Primer extension and S1 nuclease protection experiments show that two transcripts initiate 136 and 140 bases upstream from the first ATG codon in all human multidrug-resistant cell lines. The mdr1 gene is expressed in human normal kidney cells and HepG2 liver cells as a poly(A)+ RNA which starts from the same sites. Less prominent transcripts were found to initiate 155-180 bases upstream from the first ATG codon in vinblastine- or adriamycin-selected cell lines and 480-630 bases upstream in colchicine-selected cell lines. Southern hybridization analyses with different portions of a full length cDNA indicate that the human mdr1 gene encompasses at least 70 kilobases of DNA amplified in all highly multidrug-resistant cell lines. PMID- 3027055 TI - Production and characterization of antibodies to gonadotropin-releasing hormone receptor. AB - Antibodies to the gonadotropin-releasing hormone (GnRH) receptor of bovine pituitary membranes have been raised in rabbits by immunization with affinity purified receptor preparations. These antibodies did not compete with 125I labeled GnRH analog (Buserelin) for binding to the receptors but did precipitate rat and bovine solubilized receptors labeled with 125I-Buserelin. Binding of the antibodies to the receptors was also demonstrated by immunoprecipitation of 125I labeled purified receptors and photoaffinity-labeled receptors. The antibodies did not have a GnRH-like activity but rather inhibited, in a dose-dependent manner, GnRH-stimulated luteinizing hormone release from cultured rat pituitary cells. In addition, the antibodies did not inhibit luteinizing hormone release stimulated by high K+ concentration. This suggests that the antibodies recognize domains of the receptor other than the binding site of the hormone and thereby inhibit the biological response. These GnRH receptor antibodies provide a useful tool for studying GnRH receptor structure, function, localization, and biosynthesis. PMID- 3027056 TI - Accessibility and multivalency of immobilized Cibacron blue F3GA. AB - The effect of immobilized dye concentration on protein complexation was observed using zonal chromatography. A monomeric protein, octopine dehydrogenase, was retained by a single interaction to a Sepharose CL-6B column containing 11.6 mM immobilized Cibacron blue F3GA. By contrast, a tetrameric protein, lactate dehydrogenase, was retained by the same column by multiple interactions. The degree of multiple interactions was found to systematically increase with increasing immobilized dye concentration. The concentration of immobilized dye accessible to protein was found to be inversely related to the concentration of ionic components in the solvent. Zonal chromatographic measurements of free dye and unconjugated matrix suggest that increasing the concentration of ionic components promotes the adsorption of immobilized dye to the adjacent matrix surface. Such adsorption markedly affects both the capacity of an immobilized dye column and the multiplicity of its interaction with oligomeric proteins. PMID- 3027057 TI - The reaction of cytochrome oxidase with cyanide. Preparation of the rapidly reacting form and its conversion to the slowly reacting form. AB - The magnitude of the slow phase of reaction of cytochrome oxidase with cyanide has been correlated with the size of the epr signal at g' = 12. This epr signal was not found in submitochondrial particles, and significant g' = 12 epr was only observed late in the purification of solubilized enzyme. The Hartzell-Beinert procedure for the purification of cytochrome oxidase (Hartzell, C.R., and Beinert, H. (1974) Biochim. Biophys. Acta 368, 318-338) has been modified so that the purified enzyme reacts in a single rapid phase with potassium cyanide and lacks the g' = 12 epr signal. This enzyme could be converted to the slowly reacting form upon incubation at low pH and/or low enzyme concentration. No procedure for the stable reversal of the process could be found. Some physical and chemical properties of the two forms of the enzyme are compared. PMID- 3027058 TI - Characterization of a cDNA for human protein C inhibitor. A new member of the plasma serine protease inhibitor superfamily. AB - A cDNA library in lambda-phage lambda gt11 containing DNA inserts prepared from human liver mRNA was screened with monoclonal antibodies to human protein C inhibitor. Six positive clones were isolated from 6 X 10(6) phages and plaque purified. The cDNA in the phage containing the largest insert, which hybridized to a DNA probe prepared on the basis of the amino-terminal amino acid sequence of the mature inhibitor, was sequenced. This cDNA insert contained 2106 base pairs coding for a 5'-noncoding region, a 19-amino acid signal peptide, a 387-amino acid mature protein, a stop codon, and a long 3'-noncoding region of 839 base pairs. Based on the amino acid sequence of the carboxyl-terminal peptide released by cleavage of protein C inhibitor by activated protein C as well as by thrombin, the reactive site peptide bond of protein C inhibitor is Arg354-Ser355. Five potential carbohydrate-binding sites were found in the mature protein. The high homology of the amino acid sequence of protein C inhibitor to the other known inhibitors clearly demonstrates that protein C inhibitor is a member of the superfamily of serine protease inhibitors including alpha 1-antichymotrypsin, alpha 1-antitrypsin, antithrombin III, ovalbumin, and angiotensinogen. Based on the difference matrices for these proteins, we present possible phylogenetic trees for these proteins. PMID- 3027059 TI - Butyrate kinase from Clostridium acetobutylicum. AB - Crude extracts of Clostridium acetobutylicum contain a butyrate kinase of high specific activity (5.2 mumol/min/mg of protein). The enzyme has been purified 77 fold in a six-step procedure to a specific activity of 402 mumol/min/mg of protein. The purified butyrate kinase showed a single band with a molecular weight of 85,000 on nondenaturing polyacrylamide gradient gel electrophoresis. Separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated that the enzyme is a dimer of two apparently identical subunits with molecular weights of 39,000. The pH optimum for the reaction in the butyryl phosphate forming direction is 7.5, and the pI of the kinase is 5.6. The amino acid composition of the enzyme is also reported. It contains no tryptophan and is low in sulfur-containing amino acids. The kinase has a broad substrate specificity and exhibits its highest relative activities with butyrate and valerate. Butyrate kinase is rapidly inactivated at 50 degrees C in the absence of a fatty acid substrate. Although a reducing agent was required for maximum activity, treatment with several sulfhydryl-modifying agents failed to inhibit the enzyme. PMID- 3027060 TI - Structural and catalytic characteristics of Escherichia coli adenylate kinase. AB - The adk gene encoding adenylate kinase in Escherichia coli was cloned in pBR322. Adenylate kinase represented about 4% of total proteins in extracts of cells containing the pBR322:adk plasmid. This allowed preparation of more than 90% pure enzyme in a single-step purification procedure. Amino acid analysis, high performance liquid chromatography separation of trypsin digests, sequence analysis of most peptides, and determination of the N-terminal sequence of the whole protein confirmed the primary structure of E. coli adenylate kinase predicted from the nucleotide sequence of the adk gene (Brune, M., Schumann, R., and Wittinghofer, F. (1985) Nucleic Acids Res. 13, 7139-7151). 2-Nitro-5 thiocyanatobenzoic acid reacted with the single cysteine residue of E. coli adenylate kinase. The cyanylated protein was cleaved upon exposure to alkaline pH, yielding two peptides corresponding to residues 1-76 and 77-214, respectively. A mixture of purified peptides tended to reassociate, recovering both catalytic activity and binding properties for adenine nucleotides. E. coli adenylate kinase has a broader specificity for nucleoside monophosphates than does the mammalian enzyme. In addition to 2'-dAMP, other nucleoside monophosphates such as 3'-dAMP, adenine-9-beta-D-arabinofuranoside 5' monophosphate, and 7-deazaadenosine (tubercidine) 5'-monophosphate were able to replace AMP as substrate. PMID- 3027061 TI - Identification of the 37-kDa protein displaying a variable interaction with the erythroid cell membrane as glyceraldehyde-3-phosphate dehydrogenase. AB - In previous studies from this laboratory we isolated and characterized a 37-kDa protein that was associated with the membrane of erythroid cells. The polypeptide appeared to undergo a lineage-specific alteration in its interaction with the membrane during erythroid development and migrated as a family of isoelectric focusing variants during analyses on two-dimensional gels. We report here that the 37-kDa protein is homologous to the enzyme glyceraldehyde-3-phosphate dehydrogenase (EC 1.2.1.12). This conclusion was reached from the results of several experimental approaches comparing the biochemical and genetic properties of the 37-kDa protein (p37) with authentic glyceraldehyde-3-phosphate dehydrogenase. Peptide maps of highly purified p37 and glyceraldehyde-3-phosphate dehydrogenase, generated with Staphylococcus V8 protease, were identical. The nucleotide sequence of a cDNA clone encoding p37 was nearly identical to the published sequence for genes encoding glyceraldehyde-3-phosphate dehydrogenase. These results suggest that the interaction of the enzyme with the red cell membrane is more complex than previously envisioned. The existence of subpopulations of glyceraldehyde-3-phosphate dehydrogenase molecules is envisioned that exhibit different levels of enzyme activity and bind to the red cell membrane with varying affinities. PMID- 3027062 TI - The mechanism of activation of heart fructose 6-phosphate,2-kinase:fructose-2,6 bisphosphatase. AB - Partially purified fructose-6-P,2-kinase:fructose-2,6-bisphosphatase from beef heart was phosphorylated by cAMP protein kinase. The phosphorylated fructose-6 P,2-kinase shows lower Km for Fru-6-P (43 versus 105 microM) and for ATP (0.55 versus 1.3 mM) but no change in the Vmax, compared to those for unphosphorylated enzyme. There was no detectable change in Km or Vmax of fructose-2,6 bisphosphatase activity by the phosphorylation. These changes in heart fructose-6 P,2-kinase were in direct contrast to previous results for the liver isozyme in which phosphorylation led to inhibition of the kinase activity and activation of the phosphatase activity. PMID- 3027063 TI - Ethanol-induced mobilization of calcium by activation of phosphoinositide specific phospholipase C in intact hepatocytes. AB - The short-term effects of ethanol on calcium homeostasis were studied in isolated hepatocytes. Ethanol caused a rapid transient activation of phosphorylase not associated with changes in cAMP levels which peaked after 20-30 s and declined slowly over a period of 5-10 min. Maximal activation was found with 200 mM ethanol, and a significant effect was observed at 25 mM ethanol. Similar effects were induced by other organic solvents and by halothane, with more hydrophobic agents being effective at lower concentrations. In hepatocytes loaded with the intracellular calcium indicator quin2, the addition of ethanol caused a transient increase in cytosolic free calcium, with a kinetic pattern compatible with its involvement in the activation of phosphorylase. Pretreatment of the hepatocytes with phenylephrine or vasopressin to deplete the hormone-sensitive calcium pools in the cells prevented the ethanol-induced calcium mobilization. In 32P-labeled hepatocytes addition of ethanol caused a small (5-7%) decrease in the level of [32P]phosphatidylinositol 4,5-bisphosphate and a 10-15% increase in [32P]phosphatidylinositol 4-phosphate and [32P]phosphatidic acid. In hepatocytes labeled with myo-[3H]inositol, ethanol induced a 50-100% increase in the levels of inositol 1,4,5-trisphosphate, inositol 1,3,4-trisphosphate, and inositol bisphosphate. The changes in the inositol 1,4,5-trisphosphate level due to ethanol paralleled the time course of the elevation of cytosolic free calcium levels and activation of phosphorylase a. The effects of ethanol were comparable to those of a physiologic (1 nM) dose of vasopressin; however, unlike with vasopressin, the inositol phosphates and cytosolic calcium levels declined to basal levels 2 min after the addition of ethanol. These results indicate that ethanol, in common with calcium-mobilizing hormones, activates hormone-sensitive phosphoinositide-specific phospholipase C. The resulting changes in inositol 1,4,5-trisphosphate can account for the mobilization of intracellular calcium and the consequent activation of phosphorylase by ethanol. PMID- 3027064 TI - Low-angle neutron scattering analysis of Na/K-ATPase in detergent solution. AB - Purified Na/K-ATPase from guinea pig renal outer medulla has been delipidated and solubilized in Brij 58 (polyoxyethylene ether; C-16, E-20). At a concentration of 2 mg of Brij 58/mg of protein, about one-half the enzyme complement was solubilized and almost 50% of Na/K-ATPase activity was retained by the enzyme micelle complex. Guinier plots of the neutron scattering profiles yielded no evidence of heterogeneity with respect to subunit composition or the state of aggregation in the solubilized oligomers. Contrast matching with D2O used to obtain estimates of the molecular weight of the micellar form of Na/K-ATPase gave a mean value of 310,000 +/- 42,700, which corresponds to an alpha 2 beta 2 tetramer. A Stuhrmann plot of the neutron scattering data yielded an estimated radius of gyration of 67 A. The Stuhrmann plot also indicated an asymmetrical distribution of neutron scattering density. On the basis of the Stuhrmann plot parameters, the estimated molecular weight, and the radius of gyration, a low resolution model was formulated of the oligomeric unit of Na/K-ATPase. PMID- 3027065 TI - Alpha 1-adrenoceptor activation elevates cytosolic calcium in rat pinealocytes by increasing net influx. AB - The regulation of [Ca2+]i in rat pinealocytes was studied using the fluorescent indicator quin2. Pinealocyte resting [Ca2+]i was approximately 100 nM; this rapidly decreased in low Ca2+ medium (approximately 10 microM), indicating there was a high turnover of [Ca2+]i in these cells. Norepinephrine (NE, 10(-6) M) increased [Ca2+]i to approximately 350 nM within 1 min; [Ca2+]i then remained elevated for 30 min. The relative potency of adrenergic agonists was NE greater than phenylephrine much greater than isoproterenol. Phentolamine (10(-6) M) and prazosin (10(-8) M) blocked the effects of adrenergic agonists; in contrast, propranolol (10(-6) M) or yohimbine (10(-6) M) had little or no effect. These observations indicate NE acts via alpha 1-adrenoceptors to elevate [Ca2+]i. The [Ca2+]i response to NE did not occur when [Ca2+]e was reduced to approximately 10 microM by adding EGTA 5s before NE, indicating an increase in net Ca2+ influx is involved rather than mobilization of Ca2+ from intracellular stores. The effect of NE was not blocked by nifedipine (10(-6) M), which did block a K+-induced increase in [Ca2+]i, presumably involving voltage-sensitive channels. Ouabain (10(-5) M) caused a gradual increase in [Ca2+]i; this increase was not blocked by nifedipine. Together these data indicate that pinealocyte [Ca2+]i may be influenced by mechanisms regulated by alpha 1-adrenoceptors, voltage-dependent Ca2+ channels, and perhaps a Na+/Ca2+ exchange mechanism stimulated by ouabain. These studies indicate that the pinealocyte is an interesting model to use to study the adrenergic regulation of [Ca2+]i because of the rapid and prolonged changes in [Ca2+]i produced by alpha 1-adrenoceptor activation. PMID- 3027067 TI - The effect of GTP and Mg2+ on the GTPase activity and the fluorescent properties of Go. AB - The structures of the guanosine 5'O-(3-thio)triphosphate (GTP gamma S)-containing guanine nucleotide-binding regulatory proteins (G proteins) are distinct from those of the GDP-containing forms. One indication of the conformational change caused by GTP gamma S is a Mg2+-sensitive increase in the intensity of the proteins' tryptophan fluorescence (Higashijima, T., Ferguson, K.M., Sternweis, P.C., Ross, E.M., Smigel, M.D., Gilman, A.G. (1987), J. Biol. Chem., 262, 762 766). GTP causes a similar change in the fluorescence of Go, a G protein from bovine brain. When Mg2+ is also present, the increase in fluorescence is transient, and the rate of decline in the intensity of the fluorescence is the same as the rate of GTP hydrolysis by the protein. The steady-state rate of hydrolysis of GTP by Go (0.3-0.4/min) is slower than the catalytic rate of the protein (2/min), because the rate-limiting step in the reaction is the release of GDP. PMID- 3027066 TI - The ATPase core of a clathrin uncoating protein. AB - Chymotryptic digestion of bovine brain uncoating ATPase produced a 60-kDa fragment that was subsequently proteolyzed to 44 kDa. Loss of clathrin cage uncoating activity paralleled the conversion of the intact 70-kDa enzyme to the 60-kDa fragment, while clathrin binding activity was lost as the 60-kDa fragment was degraded to 44 kDa. This 44-kDa fragment has been purified to homogeneity and characterized as a clathrin-independent ATPase. The 44-kDa ATPase domain has been localized within the intact enzyme by the use of amino-terminal specific antibodies. This localization relates to the conserved nature of the 70-kDa heat shock protein family, of which bovine brain uncoating ATPase is a constitutively expressed member. PMID- 3027068 TI - On the active sites of the [NiFe] hydrogenase from Desulfovibrio gigas. Mossbauer and redox-titration studies. AB - The [NiFe] hydrogenase isolated from Desulfovibrio gigas was poised at different redox potentials and studied by Mossbauer spectroscopy. The data firmly establish that this hydrogenase contains four prosthetic groups: one nickel center, one [3Fe-xS], and two [4Fe-4S] clusters. In the native enzyme, both the nickel and the [3Fe-xS] cluster are EPR-active. At low temperature (4.2 K), the [3Fe-xS] cluster exhibits a paramagnetic Mossbauer spectrum typical for oxidized [3Fe-xS] clusters. At higher temperatures (greater than 20 K), the paramagnetic spectrum collapses into a quadrupole doublet with parameters magnitude of delta EQ magnitude of = 0.7 +/- 0.06 mm/s and delta = 0.36 +/- 0.06 mm/s, typical of high spin Fe(III). The observed isomer shift is slightly larger than those observed for the three-iron clusters in D. gigas ferredoxin II (Huynh, B. H., Moura, J. J. G., Moura, I., Kent, T. A., LeGall, J., Xavier, A. V., and Munck, E. (1980) J. Biol. Chem. 255, 3242-3244) and in Azotobacter vinelandii ferredoxin I (Emptage, M. H., Kent, T. A., Huynh, B. H., Rawlings, J., Orme-Johnson, W. H., and Munck, E. (1980) J. Biol. Chem. 255, 1793-1796) and may indicate a different iron coordination environment. When D. gigas hydrogenase is poised at potentials lower than -80 mV (versus normal hydrogen electrode), the [3Fe-xS] cluster is reduced and becomes EPR-silent. The Mossbauer data indicate that the reduced [3Fe-xS] cluster remains intact, i.e. it does not interconvert into a [4Fe-4S] cluster. Also, the electronic properties of the reduced [3Fe-xS] cluster suggest that it is magnetically isolated from the other paramagnetic centers. PMID- 3027069 TI - Isolation, complementary DNA sequence, and regulation of rat hepatic lauric acid omega-hydroxylase (cytochrome P-450LA omega). Identification of a new cytochrome P-450 gene family. AB - Lauric acid omega-hydroxylase (cytochrome P-450LA omega) was purified from livers of rats that had been given the hypolipidemic drug clofibrate. Immunoblot analysis with anti-cytochrome P-450LA omega antibody revealed the presence of two clofibrate-induced cytochrome P-450 proteins in both liver and kidney. This antibody was used to screen a lambda gt11 expression library constructed from liver mRNA isolated from rats given clofibrate. The clone, pP-450LA omega, was isolated and found to contain 2062 base pairs with an open reading frame of 509 amino acids (Mr 58,222). The pP-450LA omega cDNA insert was placed in the yeast expression vector pAAH5, and the resultant plasmid expressed a protein of the same size and activity as cytochrome P-450LA omega. The mechanism of the regulation of the cytochrome P-450LA omega gene by hypolipidemic agents was examined. The rate of cytochrome P-450LA omega gene transcription was increased as early as 1 h after administration of clofibrate, and this increase was followed by an elevation of cytochrome P-450LA omega mRNA, immunochemically detectable protein, and cytochrome P-450LA omega activity. In contrast, no increase in transcriptional activity was detected after clofibrate administration for the cytochrome P-450PCN or P-450b/e genes. The cytochrome P-450LA omega has several structural features in common with other cytochrome P-450s, including a conserved cysteine-containing heme-binding fifth ligand fragment and a hydrophobic amino terminus. Overall, cytochrome P-450LA omega shared less than 35% cDNA nucleotide and amino acid similarity with cytochromes P-450c, P-450d, P 450e, and P-450PCN, indicating that it is a member of a new cytochrome P-450 gene family. Southern blot analysis indicates that this family contains two or three genes. PMID- 3027070 TI - Differentiation of human promyelocytic HL 60 cells by retinoic acid is accompanied by an increase in the intracellular pH. The role of the Na+/H+ exchange system. AB - Retinoic acid, which induces the differentiation of HL 60 cells to granulocytes, produces a cell alkalinization from pHi = 7.03 to pHi = 7.37. The half-maximum effect of retinoic acid is observed at 10 nM. The effect of retinoic acid on the pHi develops slowly, and it precedes the differentiation of the cells. A cell alkalinization is also observed after differentiation of the cells by dimethyl sulfoxide. It is not observed using etretinate, a synthetic retinoid that does not promote the differentiation of HL 60 cells. Two pHi regulating mechanisms coexist in HL 60 cells. The Na+/H+ exchange system is the major mechanism that allows HL 60 cells to recover from an intracellular acidosis. A second mechanism is a Na-HCO3 cotransport system. During differentiation of the cells by retinoic acid, a 2-fold increase in the activity of the Na+/H+ exchange system is observed, while the activity of the NaHCO3 cotransport remains constant. The properties of interaction of the Na+/H+ exchanger with internal H+, external Na+, and Li+ as well as with amiloride and its derivatives are defined. The Na+/H+ exchanger of HL 60 cells is characterized by unusually low affinities for alkali cations and a high affinity for amiloride and its derivatives. The pHi dependence of the exchanger is not modified after differentiation by retinoic acid. It is concluded that the mechanism of activation of the Na+/H+ exchanger by retinoic acid is distinct from the short-term effect produced by mitogens and phorbol esters which change the pHi dependence of the system. PMID- 3027071 TI - A correlation between phorbol diester-induced protein phosphorylation and superoxide anion generation in HL-60 cells during granulocytic maturation. AB - As HL-60 cells matured along the granulocytic pathway, phorbol diester-induced superoxide anion production was compared to phorbol diester-induced protein phosphorylation using an in vitro phosphorylation technique. Maturation was induced by 0, 2, 4, or 6 days incubation with dimethyl sulfoxide (Me2SO). In 0 day Me2SO HL-60 cells, phorbol 12-myristate 13-acetate induced phosphorylation of protein pp29 (Mr = 28,600) and to a lesser extent protein pp76 (Mr = 76,300). With increased time of Me2SO incubation, phorbol 12-myristate 13-acetate induced phosphorylation of pp212 (Mr = 211,800), pp134 (Mr = 134,200), and pp76, whereas the phosphorylation of pp29 did not change appreciably. In close agreement with this increase in protein phosphorylation was the observed increase in phorbol diester-induced superoxide anion formation. Morphological characterization of cells during Me2SO-induced differentiation reveals that these increases in phorbol diester responses are probably attributable to the proportional rise in metamyelocytes, band, and segmented neutrophils. A variety of phorbol diesters increased superoxide anion generation in HL-60 cells differentiated into granulocyte-like cells by 6-day incubation with Me2SO. The structure-activity relationship of these phorbol diester derivatives for protein phosphorylation was strongly correlated to their ability to increase superoxide anion generation. Thus, we propose that phorbol diester-induced phosphorylation of pp212, pp134, and pp76, but not pp29 may play a role in mediating the functional response of phorbol diester-induced superoxide anion generation in HL-60 cells differentiated into mature granulocyte-like cells. PMID- 3027072 TI - Covalent modification of the renal Na+/H+ exchanger by N,N' dicyclohexylcarbodiimide. AB - We studied the effect of the carboxyl group-specific reagent N,N' dicyclohexylcarbodiimide on the Na+/H+ exchanger present in microvillus membrane vesicles isolated from rabbit renal cortices. Pretreatment of membrane vesicles with dicyclohexylcarbodiimide resulted in irreversible inhibition of Na+/H+ exchange which was not due to vesicle disruption or collapse of imposed pH gradients. Inhibition by dicyclohexylcarbodiimide followed pseudo-first-order kinetics, resulted primarily from a decrease in binding affinity for substrate, was pH-dependent in a manner consistent with reaction with carboxyl groups, and was greater than inhibition by hydrophilic carbodiimides. Substrates Na+ and Li+ and the competitive inhibitor amiloride protected against inhibition by dicyclohexylcarbodiimide in a pH-dependent fashion. Finally, we demonstrated amiloride-sensitive covalent binding of radiolabeled dicyclohexylcarbodiimide to a 100-kDa protein. In conclusion, a catalytically important carboxyl group is located in a relatively hydrophobic microenvironment at or near the external transport site of the renal Na+/H+ exchanger; and the transporter itself, or a subunit thereof, may be a 100-kDa protein. PMID- 3027073 TI - Intracellular receptors for inositol 1,4,5-trisphosphate in angiotensin II target tissues. AB - Many cells (including angiotensin II target cells) respond to external stimuli with accelerated hydrolysis of phosphatidylinositol 4,5-bisphosphate, generating 1,2-diacylglycerol and inositol 1,4,5-trisphosphate, a rapidly diffusible and potent Ca2+-mobilizing factor. Following its production at the plasma membrane level, inositol 1,4,5-trisphosphate is believed to interact with specific sites in the endoplasmic reticulum and triggers the release of stored Ca2+. Specific receptor sites for inositol 1,4,5-trisphosphate were recently identified in the bovine adrenal cortex (Baukal, A. J., Guillemette, G., Rubin, R., Spat, A., and Catt, K. J. (1985) Biochem. Biophys. Res. Commun. 133, 532-538) and have been further characterized in the adrenal cortex and other target tissues. The inositol 1,4,5-trisphosphate-binding sites are saturable and present in low concentration (104 +/- 48 fmol/mg protein) and exhibit high affinity for inositol 1,4,5-trisphosphate (Kd 1.7 +/- 0.6 nM). Their ligand specificity is illustrated by their low affinity for inositol 1,4-bisphosphate (Kd approximately 10(-7) M), inositol 1-phosphate and phytic acid (Kd approximately 10(-4) M), fructose 1,6 bisphosphate and 2,3-bisphosphoglycerate (Kd approximately 10(-3) M), with no detectable affinity for inositol 1-phosphate and myo-inositol. These binding sites are distinct from the degradative enzyme, inositol trisphosphate phosphatase, which has a much lower affinity for inositol trisphosphate (Km = 17 microM). Furthermore, submicromolar concentrations of inositol 1,4,5 trisphosphate evoked a rapid release of Ca2+ from nonmitochondrial ATP-dependent storage sites in the adrenal cortex. Specific and saturable binding sites for inositol 1,4,5-trisphosphate were also observed in the anterior pituitary (Kd = 0.87 +/- 0.31 nM, Bmax = 14.8 +/- 9.0 fmol/mg protein) and in the liver (Kd = 1.66 +/- 0.7 nM, Bmax = 147 +/- 24 fmol/mg protein). These data suggest that the binding sites described in this study are specific receptors through which inositol 1,4,5-trisphosphate mobilizes Ca2+ in target tissues for angiotensin II and other calcium-dependent hormones. PMID- 3027074 TI - Purification and properties of polycation-stimulated phosphorylase phosphatases from rabbit skeletal muscle. AB - Four types of polycation-stimulated (PCS) phosphorylase phosphatases have been isolated from rabbit skeletal muscle. They are called PCSH (390 kDa), PCSM (250 kDa), and PCSL (200 kDa) phosphatase according to the apparent molecular weight of the native enzymes in gel filtration. Two forms of PCSH phosphatase could be separated by Mono Q fast protein liquid chromatography: PCSH1 and PCSH2. In the absence of polycations, the specific activities of the PCSH1, PCSH2, PCSM, and PCSL phosphatase were 400, 680, 600, and 3000 units/mg, respectively, using phosphorylase a as a substrate. They all contain a 62-65- and a 35-kDa subunit, the latter being the catalytic subunit. In addition PCSH1 phosphatase contains a 55-kDa subunit and the PCSM phosphatase a 72-75-kDa subunit in a substoichiometric ratio. All the PCS phosphatases are insensitive to Ca2+ calmodulin, inhibitor-1, and modulator protein. They display a high specificity for the alpha-subunit of phosphorylase kinase and a broad substrate specificity. The PCSH1 and PCSH2 phosphatases, but not the catalytic subunit (PCSC phosphatase), show a high degree of specificity for the deinhibitor protein. During the purification the phosphorylase to inhibitor-1 phosphatase activity ratio (10:1) remained constant for the PCSH and PCSL enzymes but decreased for the PCSM phosphatase. The stimulation observed with low concentrations of polycations is enzyme directed. The different enzyme forms show a characteristic concentration optimum and degree of stimulation. At higher concentrations, polycations become inhibitory and a time-dependent deactivation of the phosphatases is observed. PMID- 3027075 TI - Dephosphorylation of phosphoproteins and synthetic phosphopeptides. Study of the specificity of the polycation-stimulated and MgATP-dependent phosphorylase phosphatases. AB - The substrate specificity of different forms of polycation-stimulated (PCSH, PCSL, and PCSC) phosphorylase phosphatases and of the catalytic subunit of the MgATP-dependent protein phosphatase from rabbit skeletal muscle was investigated. This was done, with phosphorylase a as the reference substrate, using the synthetic phosphopeptides patterned after the phosphorylated sites of pyruvate kinase (type L) (Arg2-Ala-Ser(32P)-Val-Ala (S2), and its Thr(32P) substitute (T4)), inhibitor-1 (Arg4-Pro-Thr(32P)-Pro-Ala (T5), Arg2-Pro-Thr(32P)-Pro-Ala (T1), and its Ser(32P) substitute (S1)), and some modified phosphopeptides (Arg2 Ala-Thr(32P)-Pro-Ala (T2) and Arg2-Pro-Thr(32P)-Val-Ala (T3)), all phosphorylated by cyclic AMP-dependent protein kinase. In addition, casein(Thr-32P), phosphorylated by casein kinase-2, was also tested. The PCS phosphatases show a striking preference for the T4 configuration, PCSC being the least efficient. The catalytic subunit of the MgATP-dependent phosphatase was almost completely inactive toward all these substrates. As shown for the PCSH phosphatase, and comparing with T4, the two proline residues flanking the Thr(P) in T1 and T5, just as in inhibitor-1, drastically imparied the dephosphorylation by lowering the Vmax and not by affecting the apparent Km. The C-terminal proline (as in T2) by itself represents a highly unfavorable factor in the dephosphorylation. The critical effect of the sequence X-Thr(P)-Pro or Pro-Thr(P)-Pro (T1, T2, T5, and inhibitor-1) can be overcome by manganese ions. The additional finding that this is not the case with the Pro-Ser(P)-Pro sequence (S1) suggests that the effect of Mn2+ is highly substrate specific. These observations show the considerable importance of the primary structure of the substrate in determining the specificity of the protein phosphatases. PMID- 3027076 TI - Purification and characterization of a myosin heavy chain kinase from Dictyostelium discoideum. AB - A Dictyostelium discoideum myosin heavy chain kinase has been purified 14,000 fold to near homogeneity. The enzyme has a Mr = 130,000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and greater than 700,000 as determined by gel filtration on Bio-Gel A-1.5m. The enzyme has a specific activity of 1 mumol/min X mg when assayed at a Dictyostelium myosin concentration of 0.3 mg/ml. A maximum of 2 mol of phosphate/mol of myosin is incorporated by the kinase, and the phosphorylated amino acid is threonine. Phosphate is incorporated only into the myosin heavy chains, not into the light chains. The actin-activated Mg2+-ATPase of Dictyostelium myosin is inhibited 70-80% following maximal phosphorylation with the kinase. The myosin heavy chain kinase requires 1 2 mM Mg2+ for activity and is most active at pH 7.0-7.5. The activity of the enzyme is not significantly altered by the presence of Ca2+, Ca2+ and calmodulin, EGTA, cAMP, or cGMP. When incubated with Mg2+ and ATP, phosphate is incorporated into the myosin heavy chain kinase, perhaps by autophosphorylation. PMID- 3027077 TI - The requirement for iron (III) in the initiation of lipid peroxidation by iron (II) and hydrogen peroxide. AB - The initiation of lipid peroxidation by Fe2+ and H2O2 (Fenton's reagent) is often proposed to be mediated by the highly reactive hydroxyl radical. Using Fe2+, H2O2, and phospholipid liposomes as a model system, we have found that lipid peroxidation, as assessed by malondialdehyde formation, is not initiated by the hydroxyl radical, but rather requires Fe3+ and Fe2+. EPR spin trapping with 5,5 dimethyl-1-pyrroline-N-oxide and the bleaching of para-nitrosodimethylaniline confirmed the generation of the hydroxyl radical in this system. Accordingly, catalase and the hydroxyl radical scavengers mannitol and benzoate efficiently inhibited the generation and the detection of hydroxyl radical. However, catalase, mannitol, and benzoate could either stimulate or inhibit lipid peroxidation. These unusual effects were found to be consistent with their ability to modulate the extent of Fe2+ oxidation by H2O2 and demonstrated that lipid peroxidation depends on the Fe3+:Fe2+ ratio, maximal initial rates occurring at 1:1. These studies suggest that the initiation of liposomal peroxidation by Fe2+ and H2O2 is mediated by an oxidant which requires both Fe3+ and Fe2+ and that the rate of the reaction is determined by the absolute Fe3+:Fe2+ ratio. PMID- 3027078 TI - Regulation of phosphoinositide phosphorylation in Swiss 3T3 cells stimulated by platelet-derived growth factor. AB - The regulation of phosphoinositide phosphorylation was studied in Swiss 3T3 cells that were stimulated by platelet-derived growth factor (PDGF). Studies with intact cells showed that the mitogen increased the incorporation of 32P into phosphatidylinositol (PtdIns), phosphatidylinositol 4-phosphate (PtdIns-P), and phosphatidylinositol 4,5-bisphosphate (PtdIns-P2) during the cell cycle, with distinct peaks of incorporation for all three phosphoinositides after 1 h, and for PtdIns and PtdIns-P2 after 20 h. Direct measurements of the activities of PtdIns kinase and PtdIns-P kinase in freeze-thawed cells revealed that the activity of PtdIns kinase was rate-limiting for the synthesis of PtdIns-P2. Maximal activities of PtdIns kinase and PtdIns-P kinase, with exogenous substrates, were unchanged during the 1st h of PDGF treatment, but doubled during the next 24 h. The increase in PtdIns kinase activity began within 2-4 h, exceeded the increase in cell protein, and was abolished by cycloheximide, which suggests that the enzyme was induced specifically in response to PDGF. The increase in activity of PtdIns-P kinase paralleled the increase in cell protein. Dose-response curves for PDGF showed that the activities of PtdIns kinase and PtdIns-P kinase at 24 h increased in proportion to the extent of mitogenic stimulation of the cells. Our results support the conclusion that the activities of PtdIns kinase and PtdIns-P kinase increase in response to PDGF, but only after several hours of cell cycle traverse. PMID- 3027079 TI - The biosynthesis of gram-negative endotoxin. A novel kinase in Escherichia coli membranes that incorporates the 4'-phosphate of lipid A. AB - Extracts of Escherichia coli contain an enzyme that generates the beta,1----6 linkage of lipid A from fatty-acylated monosaccharide precursors, according to the reaction: 2,3-diacyl-GlcN-1-P + UDP-2,3-diacyl-GlcN----2,3-diacyl-GlcN (beta, 1----6)2,3-diacyl-GlcN-1-P + UDP (Ray, B. L., Painter, G., and Raetz, C. R. H. (1984) J. Biol. Chem. 259, 4852-4859). We now describe a membrane-bound kinase that phosphorylates the 4'-position of the above tetraacyldisaccharide 1 phosphate product. The lipid A 4'-kinase is distinct from the diglyceride kinase of E. coli. When crude membrane preparations are employed, several nucleoside triphosphates are able to support the phosphorylation of the tetraacyldisaccharide 1-phosphate, but ATP is the most efficient. The 4'-kinase requires Mg2+ and is stimulated by phospholipids, especially cardiolipin. Under optimal conditions the specific activity in crude extracts is 0.5 nmol/min/mg. The enzyme is rapidly inactivated by preincubation in the presence of detergents, such as Nonidet P-40 or octylglucoside, but phosphoenolpyruvate and glycerol stabilize the enzyme. The product generated in vitro has been characterized by fast atom bombardment mass spectrometry and by 1H and 31P NMR spectroscopy. Those analyses confirm that the 4' hydroxyl is the site of phosphorylation. The 4' kinase reported here is likely to represent a key step in the de novo biosynthesis of lipid A. PMID- 3027080 TI - Properties of bovine heart mitochondrial cytochrome b560. AB - A large-scale preparation of the two-subunit protein complex (QPs) that converts succinate dehydrogenase into succinate-ubiquinone reductase from cytochrome b-c1 particles is achieved by a procedure involving Triton X-100 solubilization and calcium phosphate column chromatography at different pH values. The isolated two subunit QPs contains 25 nmol of cytochrome b560/mg of protein and is able to reconstitute with soluble succinate dehydrogenase to form a TTFA-sensitive succinate-ubiquinone reductase. The maximum reconstitutive activity is 100 mumol of succinate oxidized per min per mg of QPs protein at 23 degrees C. Although cytochrome b560 in isolated QPs is not succinate reducible and its dithionite reduced form is reactive to carbon monoxide, cytochrome b560 is shown to be physically associated with succinate dehydrogenase by the following observations. The dithionite reduced form of cytochrome b560 in isolated QPs has a symmetrical alpha-absorption peak, which upon reconstitution with succinate dehydrogenase becomes slightly broadened and shows a shoulder at around 553 nm, identical to that of cytochrome b560 in succinate-ubiquinone reductase. Upon addition of succinate dehydrogenase to QPs, about 50% of the reduced form of cytochrome b560 in the QPs becomes insensitive to carbon monoxide treatment. The redox potential of cytochrome b560 in QPs is -144 mV which is higher than that of cytochrome b560 in succinate-ubiquinone reductase (-185 mV). Upon addition of succinate dehydrogenase, the redox potential of about 46% of the cytochrome b560 in QPs preparation becomes identical to that of cytochrome b560 in succinate-ubiquinone reductase. Cytochrome b560 in the QPs preparation shows two epr signals, g = 3.07 and g = 2.92, whereas cytochrome b560 in succinate-ubiquinone reductase exhibits only one epr signal at g = 3.46. When QPs is reconstituted with succinate dehydrogenase to form succinate-ubiquinone reductase, the g = 3.46 epr signal reappears at the expense of the g = 3.07 signal. Based on epr measurement at liquid helium temperature, about 18% of the total cytochrome b in the isolated active succinate-cytochrome c reductase is cytochrome b560, indicating that cytochrome b560 is indeed a unique cytochrome b and not a denatured product of cytochrome b562 or b565. PMID- 3027081 TI - Spectroscopic and kinetic properties of an oxygen-binding heme protein from Chromatium vinosum. AB - Resonance Raman and electron paramagnetic resonance spectroscopy have been utilized to identify histidine as an axial heme ligand in a high spin, heme c containing protein isolated from the photosynthetic purple sulfur bacterium Chromatium vinosum. Resonance Raman spectroscopy has also been used to characterize the CO adduct of the C. vinosum hemoprotein. Resonance Raman spectra of the heme site obtained within 10 ns of CO photolysis from the ferrous hemoprotein are virtually identical to those of the unligated protein, indicating that there is little or no rearrangement of the heme pocket in response to ligand photolysis. The equilibrium constant for CO binding to the ferrous hemeprotein was measured to be 1.7 X 10(-5) M-1 and the CO association rate constant determined to be 5.4 X 10(3) M-1 S-1. The quantum efficiency for photodissociation of the hemoprotein X CO complex was greater than or equal to 0.9. PMID- 3027082 TI - Pre-replicative association of multiple replicative enzyme activities with the nuclear matrix during rat liver regeneration. AB - As a step toward the molecular elucidation of the putative replicational apparatus associated with the nuclear matrix, we have investigated the possible matrix association of several replicational related enzymes. In addition to the previously identified DNA polymerase alpha, DNA primase, 3'-5' exonuclease, RNase H, and DNA methylase were all recovered at significant levels (20-30% of total nuclear activity) in nuclear matrix isolated from regenerating rat liver during maximal in vivo replication (22 h post-hepatectomy). In contrast, DNA ligase was not detected on the nuclear matrix even though significant activity was present in isolated nuclei. Examination of the replicative dependency of these enzyme activities following partial hepatectomy revealed pre-replicative elevations which were distinct for each matrix-bound enzyme. A second late-replicative peak in DNA methylase is consistent with a role of this matrix-bound enzyme in the maintenance of the inheritable methylation pattern. Mild sonication resulted in a significant release of all of these activities except RNase H. A major portion of the matrix-solubilized DNA polymerase alpha, DNA primase, 3'-5' exonuclease, and DNA methylase activities cosedimented on sucrose gradients between approximately 8-12 S. Our results are consistent with the organization of at least a portion of these replicative enzymes into nuclear matrix-bound replicational complexes. We also propose a novel pre-replicative assembly model of the matrix-bound replicational apparatus in which DNA primase plays an initial and critical role. PMID- 3027083 TI - Structure, organization, and transcription of Drosophila U6 small nuclear RNA genes. AB - U6 RNA is an abundant, capped small nuclear RNA (snRNA) associated with hnRNP particles (Reddy, R., and Busch, H. (1983) Prog. Nucleic Acid Res. Mol. Biol. 30, 127-162). Small nuclear ribonucleoprotein particles containing U4 and U6 RNAs are required components for splicing of pre-mRNAs (Berget and Robberson, 1986; Black and Steitz, 1986). In this study the Drosophila U6 RNA genes have been isolated and characterized. The Drosophila genome contains three U6 snRNA genes which are clustered in a 2-kilobase-pairs long DNA fragment. The U6 RNA coding regions are 100% homologous in all three genes, but the flanking sequences diverged significantly from each other. A possible secondary structure model for the Drosophila U4/U6 RNA complex is presented. Consistent with our previous observation that U6 RNA is a RNA polymerase III product (Reddy, R., Henning, D., Das, G., Harless, M., and Wright, D. (1987) J. Biol. Chem. 262, 75-81), all three genes contained a region homologous to the consensus intragenic regulatory region and a cluster of T residues on the 3'-end, characteristic of genes transcribed by RNA polymerase III. A TATA box was found between nucleotides -23 and -31, and a stretch of 28 nucleotides from -43 to -71 was conserved in the 5'-flanking region of all three U6 RNA genes. The Drosophila U6 RNA genes were transcribed in vitro by Drosophila nuclear extracts but were not transcribed by Novikoff hepatoma or HeLa cell extracts. Similarly, a mouse U6 RNA gene was transcribed in Novikoff hepatoma or HeLa cell extracts but not in Drosophila nuclear extracts. These results suggest that species-specific factor(s) are involved in the transcription of U6 snRNA genes. PMID- 3027084 TI - Preferential synthesis of diacyl and alkenylacyl ethanolamine and choline glycerophospholipids in rabbit platelet membranes. AB - In rabbit platelet membranes, the contents of alkenylacyl phospholipids (plasmalogen) were 56% of phosphatidylethanolamine and 3% of phosphatidylcholine. This uneven distribution of plasmalogens in each phospholipid class could be attributed to the different substrate specificity of ethanolaminephosphotransferase (EC 2.7.8.1) and cholinephosphotransferase (EC 2.7.8.2). The properties of the enzymes were studied, using endogenous diglycerides and CDP-[3H]ethanolamine or CDP-[14C]choline as substrates. The newly formed phospholipids were mainly diacyl and alkenylacyl and only rarely alkylacyl type. The ratios of the labeled alkenylacyl to diacyl type of phospholipids clearly varied with the concentrations of CDP-ethanolamine or CDP choline. When 1, 10, and 30 microM CDP-[3H]ethanolamine were used, the labeled phospholipids contained 53, 37, and 27% of the alkenylacyl type, respectively. The apparent Km for CDP-ethanolamine to synthesize alkenylacyl and diacyl types were 2.2 and 8.1 microM. On the other hand, when 1, 10, and 30 microM CDP [14C]choline were used, the labeled lipids contained 10, 17, and 24% alkenylacyl type, respectively. The apparent Km for CDP-choline to synthesize alkenylacyl and diacyl types were 24 and 4.3 microM. Further, the syntheses of diacyl type of phosphatidylethanolamine and the alkenylacyl type of phosphatidylcholine were markedly inhibited by unlabeled CDP-choline and CDP-ethanolamine, respectively. The two enzymes had opposite substrate specificities, and ethanolaminephosphotransferase showed a high preference to plasmalogen synthesis, especially in the presence of CDP-choline. PMID- 3027085 TI - Chicken intestinal receptor for 1,25-dihydroxyvitamin D3. Immunologic characterization and homogeneous isolation of a 60,000-dalton protein. AB - The chick 1,25-dihydroxyvitamin D3 receptor has been identified via immunoblot analysis and isolated to homogeneity via positive immunoselection and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Cytosolic extracts of intestinal mucosa, as well as purified samples highly enriched for receptor by nonimmunologic methodology were electrophoresed on denaturing gels, transferred to nitrocellulose, and probed utilizing a purified monoclonal antibody against the chick receptor. Two protein signals were detected by this approach, a major species of 60,300 daltons and a minor form at 58,600 daltons. Both immunologically identified species were present in receptor-positive tissues but were absent in receptor-negative liver extracts. The two immunoreactive cytosolic proteins comigrated with two polypeptides detected via Coomassie Blue staining as well as by immunoblot analysis after enrichment utilizing DNA-cellulose, blue dextran-Sepharose, and other chromatographic separation techniques. Increasing concentrations of the minor form during purification suggest it arises from the larger molecular weight species via proteolysis. Finally, both forms of the receptor were isolated to near homogeneity employing positive immunoselection and each individually purified to homogeneity employing sodium dodecyl sulfate polyacrylamide gel electrophoresis. These experiments show that the chick receptor exists as a major species of 60,300 as well as a minor form of 58,600 and that both forms can be purified to homogeneity via immunoaffinity chromatography. PMID- 3027087 TI - Plectin and IFAP-300K are homologous proteins binding to microtubule-associated proteins 1 and 2 and to the 240-kilodalton subunit of spectrin. AB - Structural and functional characteristics of plectin from intermediate filament preparations of rat glioma C6 cells were compared to those of the intermediate filament-associated protein of Mr = 300,000 (IFAP-300K) of baby hamster kidney cells (Yang, H.-S., Lieska, N., Goldman, A.E., and Goldman, R.D. (1985) J. Cell Biol. 100, 620-631). After radiolabeling and proteolytic digestion under varied conditions, both proteins yielded nearly identical peptide maps. Immunological cross-reactivity, co-migration on one- and two-dimensional high-resolution gels, chromatofocusing, and amino acid analysis demonstrated structural homology as well. In vivo labeling with 32Pi showed that plectin was the target for cAMP independent protein kinases which phosphorylated 18-kDa domains at the end(s) of the molecule. Previously reported phosphorylation sites for cAMP-dependent and a newly identified site for Ca2+/calmodulin-dependent protein kinases were located on different domains. In solid-phase binding assays, plectin bound to vimentin, microtubule-associated proteins 1 and 2, the 240-kDa chain of brain fodrin, and alpha-spectrin from human erythrocytes. Similar characteristics were revealed for corresponding 300-kDa components of various other cell lines, supporting the concept that plectin is a general cytoskeletal cross-linking element, probably of multiple function. PMID- 3027086 TI - Immunochemical detection of unique proteolytic fragments of the chick 1,25 dihydroxyvitamin D3 receptor. Distinct 20-kDa DNA-binding and 45-kDa hormone binding species. AB - We have characterized proteolytic fragments of the chick intestinal 1,25 dihyroxyvitamin D3 (1,25-(OH)2D3) receptor, produced through either exogenous or endogenous protease action, utilizing a variety of physical and functional assays coupled to immunoblot detection methodology. Treatment of intestinal cytosol with increasing concentrations of trypsin resulted in a progressive diminishment of the 60-kDa receptor concomitant with the appearance of a 20-kDa fragment reactive by Western blot analysis to an anti-1,25-(OH)2D3 receptor monoclonal antibody. Cleveland analysis supported the receptor-origin of this 20-kDa fragment: a common immunoreactive species of 12 kDa could be generated by Staphylococcus aureus V8 protease treatment of the intact 60-kDa receptor as well as the 20-kDa proteolytic product. The 20-kDa fragment did not bind hormone but was capable of interacting with DNA-cellulose in a fashion identical to that of the 60-kDa receptor and, therefore, may contain the functional DNA-binding domain of the chick 1,25-(OH)2D3 receptor. Thus, this fragment likely represents the complement of a larger hormone-bound fragment that we have previously described (Allegretto, E. A., and Pike, J.W. (1985) J. Biol. Chem. 260, 10139-10145). In contrast to the exogenous effect of trypsin, incubation of cytosol resulted in the time-dependent formation of an endogenous protease-derived fragment of 45 kDa. Cleveland analysis was consistent with the 60-kDa receptor derivation of the 45-kDa fragment. This species retained the hormone-binding site and the antibody determinant but was devoid of DNA-binding activity. Moreover, it generated neither the trypsin-dependent 20-kDa fragment nor the V8 protease-dependent 12 kDa species and, therefore, was derived from the opposite end of the receptor molecule. These data have facilitated the construction of a schematic model of the chick receptor in which the immunoreactive epitope is located between the functional domains for hormone binding and DNA binding. PMID- 3027088 TI - Purification and characterization of an acid phosphatase that displays phosphotyrosyl-protein phosphatase activity from bovine cortical bone matrix. AB - An acid phosphatase activity that displayed phosphotyrosyl-protein phosphatase has been purified from bovine cortical bone matrix to apparent homogeneity. The overall yield of the enzyme activity was greater than 25%, and overall purification was approximately 2000-fold with a specific activity of 8.15 mumol of p-nitrophenyl phosphate hydrolyzed per min/mg of protein at pH 5.5 and 37 degrees C. The purified enzyme was judged to be purified based on its appearance as a single protein band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (silver staining technique). The enzyme could be classified as a band 5-type tartrate-resistant acid phosphatase isoenzyme. The apparent molecular weight of this enzyme activity was determined to be 34,600 by gel filtration and 32,500 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the presence of reducing agent, indicating that the active enzyme is a single polypeptide chain. Kinetic evaluations revealed that the acid phosphatase activity appeared to catalyze its reaction by a pseudo Uni Bi hydrolytic two-step transfer reaction mechanism and was competitively inhibited by transition state analogs of Pi. The enzyme activity was also sensitive to reducing agents and several divalent metal ions. Substrate specificity evaluation showed that this purified bovine skeletal acid phosphatase was capable of hydrolyzing nucleotide tri- and diphosphates, phosphotyrosine, and phosphotyrosyl histones, but not nucleotide monophosphates, phosphoserine, phosphothreonine, phosphoseryl histones, or low molecular weight phosphoryl esters. Further examination of the phosphotyrosyl-protein phosphatase activity indicated that the optimal pH at a fixed substrate concentration (50 nM phosphohistones) for this activity was 7.0. Kinetic analysis of the phosphotyrosyl-protein phosphatase activity indicated that the purified enzyme had an apparent Vmax of approximately 60 nmol of [32P]phosphate hydrolyzed from [32P]phosphotyrosyl histones per min/mg of protein at pH 7.0 and an apparent Km for phosphotyrosyl proteins of approximately 450 nM phosphate group. In summary, the results of these studies represent the first purification of a skeletal acid phosphatase to apparent homogeneity. Our observation that this purified bovine bone matrix acid phosphatase was able to dephosphorylate phosphotyrosyl proteins at neutral pH is consistent with our suggestion that this enzyme may function as a phosphotyrosyl-protein phosphatase in vivo. PMID- 3027089 TI - Purification and characterization of an outer membrane-bound protein involved in long-chain fatty acid transport in Escherichia coli. AB - We report the purification and localization of the fadL gene product (FLP), an essential component of the long-chain fatty acid transport machinery in Escherichia coli. FLP was extracted from total membranes by differential extraction with the nonionic detergents Tween 20 and Triton X-100. This protein was further purified from a Tween 20-insoluble-Triton X-100-soluble extract by salt fractionation, gel filtration chromatography, and hydrophobic interaction chromatography. This regime results in a 95-fold purification of FLP from total membranes. The purified protein preparation was homogeneous based on silver staining and gave the characteristic behavior established for the fadL gene product in the presence of sodium dodecyl sulfate at different temperatures prior to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (Mr of 33,000 when heated at 25 degrees C and Mr of 43,000 when heated at 100 degrees C) and on two dimensional polyacrylamide gels (pI of 4.6 and a Mr of 33,000). Purified FLP was rich in hydrophobic residues accounting for approximately 45% of the total amino acid composition. To localize FLP, antisera were raised against the purified protein and were used to probe differentially fractionated membranes by Western immunoblotting. This procedure demonstrated the presence of this protein only in the outer membrane fraction of fadL+ strains. We confirmed the outer membrane localization of FLP by measuring long-chain fatty acid transport in fadL+ and fadL strains treated with EDTA to alter outer membrane permeability and in spheroplasts generated from fadL+ and fadL strains. Both EDTA-treated cells and spheroplasts transported long-chain fatty acids at essentially the same rate regardless of whether they contained a wild-type or mutant fadL gene. These data imply that FLP is a protein in the outer membrane which is specifically involved in long-chain fatty acid transport. PMID- 3027090 TI - Amino-terminal deletions in the presequence of an imported mitochondrial protein block the targeting function and proteolytic cleavage of the presequence at the carboxy terminus. AB - Subunit IV of yeast cytochrome oxidase is made in the cytosol with a 25-residue presequence. This presequence targets subunit IV into mitochondria and is removed by a protease in the matrix space. Here we show that removal of as few as 4 amino terminal residues from the subunit IV presequence (which had been attached to the cytosolic protein dihydrofolate reductase) blocks import of the protein into mitochondria and proteolytic removal of the presequence by the soluble matrix protease. Thus, this protease requires not only an appropriate cleavage site at the carboxy-terminal end of the presequence, but also information at the extreme amino terminus of the presequence. PMID- 3027091 TI - Isolation and characterization of the rat thyrotropin beta-subunit gene. Differential regulation of two transcriptional start sites by thyroid hormone. AB - The gene encoding the beta-subunit of rat thyrotropin (TSH beta) has been isolated and characterized. Blot hybridization of restriction enzyme digests of rat genomic DNA suggests that the gene is present in a single copy. The transcriptional unit is 4.9 kilobases in size representing 3 exons interrupted by 2 introns of 3.9 and 0.4 kilobases. Its nucleotide sequence reveals that the locations of the exon/intron junctions are one nucleotide upstream from the translational start and between codons +34 and +35. The location of the second intron is apparently strictly conserved among the glycoprotein hormone beta subunit genes being four codons downstream from a region encoding a consensus sequence: Cys-Ala-Gly-Tyr. Using S1 nuclease mapping and oligonucleotide-primed reverse transcription of normal and thyroidectomized rat pituitary mRNA, two transcriptional start sites were identified in the rat TSH beta gene that are 28 and 71 nucleotides upstream from the translational start site. The level of TSH beta mRNA containing the downstream site is altered by thyroidal status whereas the other mRNA utilizing the upstream cap site appears to be constitutively expressed. Characteristic promoter elements are present in the 5'-flanking region including TATAAA or Goldberg-Hogness consensus regions which are present 29 and 26 bases upstream from the respective starts of transcription. Also, several CAAT boxlike sequences are located between 95 and 300 bases upstream from the start of translation. Isolation and characterization of the gene encoding the TSH beta gene will facilitate the study of the molecular mechanisms by which hormones regulate TSH beta gene expression. PMID- 3027092 TI - Evidence for a ferryl Fea3 in oxygenated cytochrome c oxidase. AB - Evidence is reported which shows that a reactive ferryl Fea3/cupric CuB binuclear couple is present at the dioxygen reduction site in "oxygenated" cytochrome c oxidase; when the fully reduced enzyme is reoxidized at low temperatures; and when partially reduced cytochrome c oxidase is reoxidized with dioxygen at room temperature. PMID- 3027093 TI - Amino acid sequence of rhizopuspepsin isozyme pI 5. AB - The complete amino acid sequence of an aspartic protease from Rhizopus chinensis, rhizopuspepsin isozyme pI 5, has been determined. Partial sequences were first obtained from the isolated isozyme by a combination of chemical and proteolytic enzyme cleavages, peptide purifications, and Edman degradations. About one-half of the sequence was revealed by this approach. To complete the amino acid sequence, a cDNA library of R. chinensis in pBR322 was constructed. An oligonucleotide probe was synthesized based on the sequence Trp-Trp-Gly-Ile-Thr, and about 40 positive clones were identified by colony hybridization. A clone, 33E2, which had an insert size of about 1.1 kilobase pairs, was found to contain the entire coding region of rhizopuspepsin isozyme pI 5. The sequence of rhizopuspepsin contains 325 amino acid residues. The alignment of the rhizopuspepsin sequence against other aspartic proteases revealed expected homology, with the closest similarity to penicillopepsin which shares 39% identical residues. Porcine pepsin shares about 36% identical residues with rhizopuspepsin. PMID- 3027094 TI - Thyrotropin regulation of membrane lipid fluidity in the FRTL-5 thyroid cell line. Its relationship to cell growth and functional activity. AB - The mitogenic effect of thyrotropin on functional rat thyroid cells of the line FRTL-5 is correlated with membrane lipid fluidity as evaluated by fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene. Continued exposure of FRTL-5 cells to a medium lacking thyrotropin causes cessation of cell proliferation and a decrease in membrane lipid fluidity which reaches its minimum in approximately 8 days. The change in lipid fluidity is due to an absolute increase (greater than 2 fold) of membrane cholesterol, with an increased cholesterol/phospholipid ratio and an increased ratio of saturated to unsaturated fatty acids of the membrane phospholipids, contributed primarily by a nearly 4-fold increase in the ratio of saturated to unsaturated C16 fatty acids. It is also associated with a variation of the relative proportions of the major membrane phospholipids; thus, phosphatidylinositol and phosphatidylethanolamine decrease while phosphatidylcholine increases. Both membrane fluidity and lipid composition can be restored by thyrotropin to their original levels, i.e. levels measured under continuous exposure to the hormone. Complete reversal requires at least 48 h, i.e. approximately the same time required for resumption of growth when FRTL-5 cells, starved in thyrotropin, are re-exposed to the hormone. Changes in lipid composition and fluidity can be prevented or can be reversed if FRTL-5 cells are exposed to dibutyryl cAMP while being deprived of thyrotropin. Dibutyryl cAMP has only a modest direct effect on growth; however, this pretreatment eliminates the 48-h lag phase with respect to thyrotropin stimulation. It is proposed that the effects of thyrotropin on growth of FRTL-5 cells requires a modification of the molecular structure and the physical state of cell membranes, which can be mediated by cAMP, although cAMP is not sufficient by itself to promote growth. PMID- 3027095 TI - Biochemical characterization of hepatic microsomal leukotriene B4 hydroxylases. AB - omega-Hydroxylation of leukotriene B4 (LTB4) has been reported in human and rodent polymorphonuclear leukocytes; preliminary information indicates that this metabolism is cytochrome P-450 dependent. Therefore, these studies were initiated to characterize the cytochrome P-450-dependent metabolism of LTB4 in other tissues. LTB4 was metabolized by rat hepatic microsomes to two products, 20 hydroxy(omega)-LTB4 and 19-hydroxy(omega-1)-LTB4. The formation of these metabolites was both oxygen and NADPH dependent indicating that a monooxygenase(s) was responsible for these reactions. The apparent Km and Vmax for LTB4 omega-hydroxylase were 40.28 microM and 1202 pmol/min/mg of protein, respectively. In contrast, the apparent Km and Vmax for LTB4 (omega-1) hydroxylase were 61.52 microM and 73.50 pmol/min/mg of protein, respectively. Both LTB4 omega- and (omega-1)-hydroxylases were inhibited by metyrapone in a concentration-dependent fashion. However, SK&F 525A inhibited LTB4 (omega-1)- but not omega-hydroxylase. In contrast, alpha-naphthoflavone decreased LTB4 omega- but not (omega-1)-hydroxylase activities. The differences in the Km apparent for substrate as well as the differential inhibition by inhibitors of cytochrome P 450 suggest that the omega- and (omega-1)-hydroxylations of LTB4 in hepatic microsomes are mediated by different isozymes of P-450. Furthermore, several additional characteristics of LTB4 hydroxylases indicate that these isozymes of P 450 may be different from those which catalyze similar reactions on medium-chain fatty acids, such as laurate and prostaglandins. PMID- 3027096 TI - Signal recognition particle arrests elongation of nascent secretory and membrane proteins at multiple sites in a transient manner. AB - The signal recognition particle (SRP) has been shown to target nascent secretory and membrane proteins to the endoplasmic reticulum. In the wheat germ cell-free system, SRP arrests the elongation of the nascent chains until the translational complex is docked to the endoplasmic reticulum membrane where the interaction between SRP and docking protein causes a release of the nascent chain arrest. For two secretory proteins, arrested peptides of 70 amino acids have been identified (Walter, P., Ibrahimi, I., and Blobel, G. (1981) J. Cell Biol. 91, 545-550; Meyer, D. I., Krause, E., and Dobberstein, B. (1982) Nature 297, 647-650). By using an in vitro coupled transcription-translation system, we have analyzed SRP arrest and the resulting peptides of the two secretory proteins lysozyme and granulocyte-macrophage colony-stimulating factor and the membrane protein invariant chain. SRP arrested the elongation of all three proteins at multiple sites, giving rise to ladders of arrested peptides. The size of the arrested peptides increased with the time of translation, resulting in mostly full-length pre-peptides after about 40 min. This suggests that SRP arrest in transient rather than stable. Upon addition of microsomes, the SRP arrest was released, and all the blocked peptides could be chased into mature proteins or full-length precursors. PMID- 3027097 TI - Regulation of neutrophil NADPH oxidase activation in a cell-free system by guanine nucleotides and fluoride. Evidence for participation of a pertussis and cholera toxin-insensitive G protein. AB - Guanine nucleotide-binding regulatory proteins (G proteins) transduce a remarkably diverse group of extracellular signals to a relatively limited number of intracellular target enzymes. In the neutrophil, transduction of the signal following fMet-Leu-Phe receptor-ligand interaction is mediated by a pertussis toxin substrate (Gi) that activates inositol-specific phospholipase C. We have utilized a plasma membrane-containing fraction from unstimulated human neutrophils as the target enzyme to explore the role of G proteins in arachidonate and cytosolic cofactor-dependent activation of the NADPH-dependent O 2-generating oxidase. When certain guanine nucleotides or their nonhydrolyzable analogues were present during arachidonate and cytosolic cofactor-dependent activation, they exerted substantial dose-dependent effects. The GTP analogue, GTP gamma S, caused a 2-fold increase in NADPH oxidase activation (half-maximal stimulation, 1.1 microM). Either GDP or its nonhydrolyzable analogue, GDP beta S, inhibited up to 80% of the basal NADPH oxidase activation (Ki GDP = 0.12 mM, GDP beta S = 0.23 mM). GTP caused only slight and variable stimulation, whereas F-, an agent known to promote the active conformation of G proteins, caused a 1.6 fold stimulation of NADPH oxidase activation. NADPH oxidase activation in the cell-free system was absolutely and specifically dependent on Mg2+. Although O2- production in response to fMet-Leu-Phe was inhibited greater than 90% in neutrophils pretreated with pertussis toxin, cytosolic cofactor and target oxidase membranes from neutrophils treated with pertussis toxin showed no change in basal- or GTP gamma S-stimulated NADPH oxidase activation. Cholera toxin treatment of neutrophils also had no effect on the cell-free activation system. Our results suggest a role for a G protein that is distinct from Gs or Gi in the arachidonate and cytosolic cofactor-dependent NADPH oxidase cell-free activation system. PMID- 3027098 TI - An in vitro study of the translational attenuation model of ermC regulation. AB - We have used a Bacillus subtilis in vitro translation system to test the translational attenuation model for ermC regulation. The ermC gene product is known to methylate rRNA, rendering ribosomes unable to bind this antibiotic. We have shown that the induction of ermC methylase in vitro is post-transcriptional and specific for the macrolides erythromycin and oleandomycin. Erythromycin has no significant effect on the stability of the ermC transcript in vitro, and hence the post-transcriptional induction of methylase under these conditions occurs by stimulation of translation. The induction effect requires ribosomes able to bind erythromycin. By adding small proportions of unmethylated to a methylated extract in the presence of erythromycin, methylase synthesis could be induced. Conversely, when small amounts of methylated extracts were mixed with unmethylated extracts, methylase synthesis could be maintained at elevated levels in the presence of a high concentration of erythromycin. These effects were specific for the inducible ermC, were not observed with a constitutive variant, and could be explained satisfactorily by the translational attenuation model. The roles of three segments of the ermC leader in regulation were explored by probing with appropriate complementary synthetic oligodeoxynucleotides. The induction effect of erythromycin was mimicked by using an oligonucleotide that could free the ribosome binding site for methylase. PMID- 3027099 TI - Demonstration of erythromycin-dependent stalling of ribosomes on the ermC leader transcript. AB - ermC encodes a methylase that modifies 23 S rRNA, conferring resistance to macrolide-lincosamide-streptogramin B antibiotics. The expression of this gene is induced by erythromycin using a translational mechanism. We have employed the inherent RNase activity of a Bacillus subtilis S-30 extract as a probe for studying the interaction of ribosomes with ermC mRNA in the presence of antibiotics. 5' end-labeled ermC runoff transcript is a substrate for this RNase activity, while the ribosome-bound region of the RNA appears to be protected. Erythromycin- and oleandomycin-dependent protection of fragments of length 79-81 was observed during the translation of end-labeled ermC transcript. This occurs only using unmethylated (erythromycin sensitive) ribosomes. Various other antibiotics including clindomycin, tylosin, and lincomycin do not show this specific protection. These effects parallel the in vivo specificity of ermC induction. The effect of erythromycin can be abolished by using oligonucleotides complementary to regions of the ermC transcript upstream from nucleotide 71 and not by using an oligonucleotide complementary to a region of ermC downstream from that position. These results are interpretable in terms of the translational attenuation model and demonstrate that erythromycin-bound ribosomes initiate translation of the leader peptide, stall upstream from nucleotide 80 on the ermC mRNA, and thus make the ribosome-binding site for methylase message available for ribosome interaction. PMID- 3027100 TI - Nucleotide sequence and hormonal regulation of mouse glycerophosphate dehydrogenase mRNA during adipocyte and muscle cell differentiation. AB - We have studied the structure and regulation of glycerophosphate dehydrogenase (GPD) mRNA during mouse adipocyte and muscle cell differentiation. This message has a size of 2.8 kilobases that includes a coding segment of 1050 bases and a large untranslated 3' end of about 1700 bases. There is a high degree of amino acid homology (91%) between the mouse adipocyte and rabbit muscle GPD proteins. GPD mRNA is not detected in myoblasts, but, as in adipogenesis, it is expressed upon differentiation into myotubes. The modulation of GPD mRNA by cyclic AMP analogues and tumor necrosis factor has been examined in both adipocytes and myotubes. Dibutyryl cAMP or 8-bromo-cAMP causes a large reduction of GPD mRNA levels in both cell types, with less than 20% remaining after 18 h of treatment. Tumor necrosis factor effects a dramatic and rapid reduction in GPD mRNA in fat cells and a slower but significant decrease in the level of this mRNA in muscle cells. These results indicate that GPD gene expression is linked to cell differentiation in both fat and muscle cells, and suggest certain similarities in hormonal modulation in both cell types. PMID- 3027101 TI - Characterization of the fusion of enveloped viruses with the plasma membrane of Saccharomyces cerevisiae spheroplasts. AB - Vesicular stomatitis virus (VSV) was associated at low pH with Saccharomyces cerevisiae spheroplasts. In the cold, the association was characterized as reversible binding to the spheroplast surface. At 37 degrees C, the association became irreversible due to fusion of the viral envelope with the yeast plasma membrane according to the following data. Proteinase K digestion degraded the viral envelope glycoprotein G but left the internal N and M proteins of VSV intact and associated with the spheroplasts. The plasma membrane could be stained by indirect immunofluorescent labeling using antiserum against VSV. By immunoelectron microscopy, no VSV particles could be detected at the spheroplast surface. Instead, the G protein could be visualized at the external aspect of the plasma membrane using specific antiserum and protein A-gold. Fusion of VSV with spheroplasts occurred below pH 4.75 at temperatures of 30-42 degrees C. It was strictly dependent on the prior removal of the yeast cell wall. The fusion process was fast, calcium-independent, and nonleaky, leaving the spheroplasts viable for at least 4 h. On the average, less than 100 VSV particles could be fused per one spheroplast. Similar data were obtained with Semliki Forest virus. PMID- 3027102 TI - Expression of porcine pro-opiomelanocortin cDNA in heterologous monkey kidney cells. Biosynthesis and secretion of the prohormone without processing. AB - Pro-opiomelanocortin is the common precursor to several pituitary hormones. These include alpha-melanotropic hormone, adrenocorticotropic hormone, beta-lipotropic hormone, and beta-endorphin. The porcine pro-opiomelanocortin cDNA was inserted downstream from the early promoter of a SV40-derived expression vector and transfected into the monkey kidney COS-1 cells. Transient expression of the pro opiomelanocortin cDNA was observed between 48 and 70 h after transfection. Analysis of pro-opiomelanocortin-related material in COS-1 cell extracts and culture medium revealed that these cells synthesize and secrete constitutively pro-opiomelanocortin without further processing into its mature hormones. Our results suggest that COS-1 cells do not contain the necessary enzymatic machinery to process complex precursors such as pro-opiomelanocortin. PMID- 3027103 TI - A vesicular intermediate in the transport of hepatoma secretory proteins from the rough endoplasmic reticulum to the Golgi complex. AB - We have identified a vesicle fraction that contains alpha 1-antitrypsin and other human HepG2 hepatoma secretory proteins en route from the rough endoplasmic reticulum (RER) to the cis face of the Golgi complex. [35S]Methionine pulse labeled cells were chased for various periods of time, and then a postnuclear supernatant fraction was resolved on a shallow sucrose-D2O gradient. This intermediate fraction has a density lighter than RER or Golgi vesicles. Most alpha 1-antitrypsin in this fraction (P1) bears N-linked oligosaccharides of composition similar to that of alpha 1-antitrypsin within the RER; mainly Man8GlcNac2 with lesser amounts of Man7GlcNac2 and Man9GlcNac2; this suggests that the protein has not yet reacted with alpha-mannosidase-I on the cis face of the Golgi complex. This light vesicle species is the first post-ER fraction to be filled by labeled alpha 1-antitrypsin after a short chase, and newly made secretory proteins enter this compartment in proportion to their rate of exit from the RER and their rate of secretion from the cells: alpha 1-antitrypsin and albumin faster than preC3 and alpha 1-antichymotrypsin, faster, in turn, then transferrin. Deoxynojirimycin, a drug that blocks removal of glucose residues from alpha 1-antitrypsin in the RER and blocks its intracellular maturation, also blocks its appearance in this intermediate compartment. Upon further chase of the cells, we detect sequential maturation of alpha 1-antitrypsin to two other intracellular forms: first, P2, a form that has the same gel mobility as P1 but that bears an endoglycosidase H-resistant oligosaccharide and is found in a compartment--probably the medial Golgi complex--of density higher than that of the intermediate that contains P1; and second, the mature sialylated form of alpha 1-antitrypsin. PMID- 3027104 TI - Regulation of amino acid transport in isolated rat hepatocytes during development. AB - The effect of amino acid depletion or supplementation and the effect of glucagon and insulin on the amino acid transport mediated by system A were investigated by determining the uptake of either 2-amino [1-14C]isobutyric acid (AIB) or N-methyl 2-amino [1-14C]isobutyric acid (MeAIB) in rat hepatocytes, freshly isolated at different stages of pre- and postnatal development. The data obtained show that the Na+-dependent uptake was higher at the earliest developmental stages, and steadily decreased until the adult level. The hormones increased AlB and MeAIB uptake enhancing the Vmax, while the Km was unchanged. This effect was evident in cells from adult and 18-20-day-old fetuses, while no response was present before the 18th day of fetal life and in the perinatal period. Actinomycin D or cycloheximide abolished this hormone-dependent increase. A decrease in AlB and MeAIB transport after incubation in an amino acid-rich medium was demonstrated at all ages tested, but was particularly evident in the prenatal life. The increase in the activity of the system following amino acid starvation was shown to be mostly dependent from de novo protein synthesis in the fetal life; on the contrary in the adult the increase appeared to be more linked to the release from transinhibition of the transport. PMID- 3027105 TI - Altered type I protein kinase in adhesion defective CHO cell variants. AB - ADv cells are Chinese hamster ovary (CHO) cell variants which cannot adhere to fibronectin coated substrata (Harper & Juliano: J. Cell Biol., 1980; Nature 1981a,b). We have shown that the defect in some clones of ADv cells is distal to the initial interaction between fibronectin and its cell surface receptors (Cheung and Juliano: Exp. Cell Res., 1984), and that it extends to fibronectin mediated aggregation and endocytosis. The adhesion defect in some ADv clones can be corrected by raising intracellular cAMP levels (Cheung & Juliano: J. Cell Physiol., 1985). Here we examine the protein kinase activities and phosphorylation patterns in an adhesion defective variant clone ADv F11CA11. Analysis of the cAMP dependent protein kinase activity (cAdPK) in crude extracts of F11CA11 cells shows an apparent increase in K (activation) as compared to wild type (WT) CHO cell extracts. Further, the DE-52 cellulose chromatography profile of cAdPK in the F11CA11 variant is markedly different from WT in that the type I cAdPK peak elicited by 1 microM cAMP is essentially missing in F11CA11, while the type II cAdPK peak is similar to that in WT. Raising the cAMP level to 100 microM elicits a type I peak in F11CA11 with about 45% of the activity of the WT peak. Binding studies with 3H-cAMP reveal that the type I peak in F11CA11 has a Kd of 1.7 X 10(-8) M as compared to 2.0 X 10(-9) M for WT, whereas the type II peak Kd is approximately 1 X 10(-9) M for both WT and F11CA11. Two-dimensional polyacrylamide gel analysis of 32Pi labeled WT cells and F11CA11 cells with or without cAMP treatment reveals the presence of a protein(s) of 50 kilodaltons which is phosphorylated in WT cells and in cAMP treated F11CA11 cells but not in untreated F11CA11 cells. These findings, coupled with our previous observations, strongly indicate that the adhesion defect in ADvF11CA11 cells is associated with an altered type I cAdPK having lower affinity for cAMP. At normal cellular cAMP levels this enzyme fails to phosphorylate one or more critical protein substrates; however, by raising internal cAMP levels, the defect can be overcome. Thus type I cAdPK seems to play an important role in the regulation of fibronectin mediated cell adhesion, cell aggregation, and endocytosis. PMID- 3027107 TI - Characterization of beta-adrenergic receptors throughout the replicative life span of IMR-90 cells. AB - Beta-adrenergic receptor number and receptor affinity for isoproterenol were assessed at various in vitro ages of the human diploid fibroblast cell line IMR 90. From population doubling level (PDL) 33 to 44, there was a positive correlation between beta-adrenergic receptor density and PDL (r = 0.709). Beta adrenergic receptors, assessed by Scatchard analysis of [125I]-iodocyanopindolol (ICYP) binding, increased from 15 fmol/mg protein at PDL 33 to 36 fmol/mg protein at PDL 44. In contrast, from PDL 44 to 59, there was a negative correlation between beta-adrenergic receptor density and PDL (r = 0.768). Receptor density declined to 12 fmol/mg protein at PDL 59. When the density of beta-adrenergic receptors was expressed as receptor per cell, the findings were similar. Receptor agonist affinity for isoproterenol was determined from Hill plots of [125I]-ICYP competition with isoproterenol. There was no change in the dissociation constant for isoproterenol with in vitro age. In humans, serum norepinephrine concentrations increase with age. This increase in serum norepinephrine may be partially responsible for the decreased beta-adrenergic receptor-agonist affinity observed with age in human lymphocytes and rat heart and lung. Similar changes in receptor-agonist affinity are observed in rat heart and human lymphocytes following exposure to beta-agonists and are part of the desensitization process. The present findings are consistent with the hypothesis that the decreases in receptor agonist affinity in rat and man with age are secondary to increases in catecholamine concentrations. PMID- 3027106 TI - Spermidine mediates degradation of ornithine decarboxylase by a non-lysosomal, ubiquitin-independent mechanism. AB - The mechanism of spermidine-induced ornithine decarboxylase (ODC, E.C. 4.1.1.17) inactivation was investigated using Chinese hamster ovary (CHO) cells, maintained in serum-free medium, which display a stabilization of ODC owing to the lack of accumulation of putrescine and spermidine (Glass and Gerner: Biochem. J., 236:351 357, 1986; Sertich et al.: J. Cell Physiol., 127:114-120, 1986). Treatment of cells with 10 microM exogenous spermidine leads to rapid decay of ODC activity accompanied by a parallel decrease in enzyme protein. Analysis of the decay of [35S]methionine-labeled ODC and separation by two-dimensional electrophoresis revealed no detectable modification in ODC structure during enhanced degradation. Spermidine-mediated inactivation of ODC occurred in a temperature-dependent manner exhibiting pseudo-first-order kinetics over a temperature range of 22-37 degrees C. In cultures treated continuously, an initial lag was observed after treatment with spermidine, followed by a rapid decline in activity as an apparent critical concentration of intracellular spermidine was achieved. Treating cells at 22 degrees C for 3 hours with 10 microM spermidine, followed by removal of exogenous polyamine, and then shifting to varying temperatures, resulted in rates of ODC inactivation identical with that determined with a continuous treatment. Arrhenius analysis showed that polyamine mediated inactivation of ODC occurred with an activation energy of approximately 16 kcal/mol. Treatment of cells with lysosomotrophic agents (NH4Cl, chloroquine, antipain, leupeptin, chymostatin) had no effect on ODC degradation. ODC turnover was not dependent on ubiquitin dependent proteolysis. Shift of ts85 cells, a temperature-sensitive mutant for ubiquitin conjugation, to 39 degrees C (nonpermissive for ubiquitin-dependent proteolysis) followed by addition of spermidine led to a rapid decline in ODC activity, with a rate similar to that seen at 32 degrees C (the permissive temperature). In contrast, spermidine-mediated ODC degradation was substantially decreased by inhibitors of protein synthesis (cycloheximide, emetine, and puromycin). These data support the hypothesis that spermidine regulates ODC degradation via a mechanism requiring new protein synthesis, and that this occurs via a non-lysosomal, ubiquitin-independent pathway. PMID- 3027108 TI - Induction of DNA synthesis in dog thyrocytes in primary culture: synergistic effects of thyrotropin and cyclic AMP with epidermal growth factor and insulin. AB - We have investigated the growth effects of thyrotropin (TSH) (mimicked by forskolin and acting through cyclic AMP), epidermal growth factor (EGF), serum (10%) and insulin on quiescent dog thyroid epithelial cells in primary culture in a serum-free defined medium. These cells were previously shown to retain the capacity to express major thyroid differentiation markers. In the presence of insulin and after a similar prereplicative phase of 18 +/- 2h, TSH, EGF, and serum promoted DNA synthesis in such quiescent cells only a minority of which had proliferated in vitro before stimulation. The combination of these factors induced more than 90% of the cells to enter S phase within 48 h and near exponetial proliferation. Analysis of the cell cycle parameters of the stimulated cells revealed that the G1 period duration was similar to the length of the prereplicative phase of quiescent thyroid cells; this might indicate that they were in fact in an early G1 stage rather than in G0 prior to stimulation. TSH and EGF action depended on or was potentiated by insulin. Strikingly, nanomolar concentrations of insulin were sufficient to support stimulation of DNA synthesis by TSH, while micromolar concentrations of insulin were required for the action of EGF. This suggests that insulin supported the action of TSH by acting on its own high affinity receptors, whereas its effect on EGF action would be related to its somatomedinlike effects at high supraphysiological concentrations. Insulin stimulated the progression in the prereplicative phase initiated by TSH or forskolin. In addition, in some primary cultures TSH must act together with insulin to stimulate early events of the prereplicative phase. In the presence of insulin, EGF, and forskolin, an adenylate cyclase activator, markedly synergized to induce DNA synthesis. Addition of forskolin 24 h after EGF or EGF 24 h after forskolin also resulted in amplification of the growth response but with a lag equal to the prereplicative period observed with the single compound. This indicates that events induced by the second factor can no longer be integrated during the prereplicative phase set by the first factor. These findings demonstrate the importance of synergistic cooperation between hormones and growth factors for the induction of DNA synthesis in epithelial thyroid cells and support the proposal that essentially different mitogenic pathways--cyclic AMP dependent or independent--may coexist in one cell. PMID- 3027109 TI - Properties of colony-stimulating factors produced by macrophage cell lines and hybrid cells. AB - Colony-stimulating factors (CSFs) produced by two simian virus 40(SV40) transformed macrophage cell lines (BAM1 and BAM3), and three hybrids (HM3-11, HM3 12, and HM3-14) derived from fusion between BAM3 and a Chinese hamster cell line (hs222-16) were examined. HM3-11 and HM3-14 produce two molecular species of CSF, which are not found in the conditioned media from cultures of BAM1 and BAM3 or lipopolysaccharide (LPS), phorbolmyristate-acetate (PMA), and zymosan-stimulated BAM3. HM3-12, which is classified into another group in terms of CSF secretion, does not produce these two CSFs. On the basis of various criteria, one of these CSF species (peak 1-CSF) was characterized as a macrophage-colony-stimulating factor (M-CSF). The other CSF (peak 2-CSF) induced a group of bone marrow cells in granulocytes and macrophages as well as growth of a mast cell line, IC2. This CSF has an apparent molecular weight of 18,000, estimated by SDS-polyacrylamide gel electrophoresis. Unlike interleukin 3 (IL3) from WEHI-3 cells, the growth factor activity of peak 2-CSF binds to DEAE-Sephacel. Thus, peak 2-CSF is similar to a granulocyte-macrophage colony-stimulating factor (GM-CSF) rather than to IL3. The anti L cell CSF serum does not inhibit the CSF activity in Chinese hamster fibroblast conditioned medium, and the IC2 cells do not respond to Chinese hamster lung conditioned medium (CHLCM), suggesting that peak 1- and peak 2-CSF are of mouse origin. PMID- 3027110 TI - Characterization and partial purification of a plasma membrane receptor for Bacillus thuringiensis var. kurstaki lepidopteran-specific delta-endotoxin. AB - The lepidopteran-specific P1 delta-endotoxin of Bacillus thuringiensis var. kurstaki HD-1 was activated in vitro using insect gut proteases and found to be highly specific for the lepidopteran cell line Choristoneura fumiferana CF1 among a wide range of lepidopteran and dipteran cell lines tested. The toxicity of P1 against CF1 cells is inhibited by N-acetylgalactosamine (GalNAc), and the lectins soybean agglutinin (SBA) and wheat-germ agglutinin. Protein blotting was used to identify a glycoprotein of 146 X 10(3) Mr in the plasma membrane of CF1 cells, capable of binding both the toxin and SBA, which is specific for GalNAc. This glycoprotein was labelled using galactose oxidase and sodium boro-[3H]hydride and solubilized in Triton X-100 before partial purification by affinity chromatography on SBA-agarose. We propose that this glycoprotein is a good candidate for the cellular receptor of the lepidopteran-specific P1 delta endotoxin of B. thuringiensis var. kurstaki HD-1. PMID- 3027112 TI - GABA and benzodiazepine receptors in the gerbil brain after transient ischemia: demonstration by quantitative receptor autoradiography. AB - Quantitative receptor autoradiography was used to measure the binding of gamma aminobutyric acid (GABA) and benzodiazepine receptors after ischemia by means of transient occlusion of bilateral common carotid arteries in the gerbil. [3H]Muscimol was used to label the GABAA receptors and [3H]flunitrazepam to label central type benzodiazepine receptors. In the superolateral convexities of the frontal cortices, [3H]muscimol binding was increased in 60% of the animals killed 3 days after ischemia, and decreased in 67% of the animals killed 27 days after ischemia. Twenty-seven days after ischemia, [3H]flunitrazepam binding in the substantia nigra pars reticulata increased to 252% of the control, though the increase in [3H]muscimol binding was not significant. In the dorsolateral region of the caudate putamen, marked neuronal necrosis and depletion of both [3H]muscimol and [3H]flunitrazepam binding sites were observed 27 days after ischemia, the ventromedial region being left intact. In spite of the depletion of pyramidal cells in the CA1 region of the hippocampus, both [3H]muscimol and [3H]flunitrazepam binding sites were preserved 27 days after ischemia. Since our previous study revealed that adenosine A1 binding sites were depleted in the CA1 subfield of the hippocampus after ischemia correlating with neuronal damage, GABAA and benzodiazepine receptors may not be distributed predominantly on the pyramidal cells in the CA1 region. PMID- 3027111 TI - Structurally related Bacillus thuringiensis delta-endotoxins display major differences in insecticidal activity in vivo and in vitro. AB - Many strains within the 22 serotypes of Bacillus thuringiensis produce crystal delta-endotoxins with slight differences in their insecticidal toxicity spectrum in vivo. Since the basis of this specificity is unknown, we chose to compare the activity of delta-endotoxins from three strains: B. thuringiensis var. kurstaki HD-1, var. aizawai HD-249 and var. thuringiensis HD-350, both in vivo and on insect cell lines in vitro. Immunoblotting with antisera to activated var. kurstaki P1 lepidopteran toxin revealed antigenic cross-reaction with the 130 X 10(3) Mr toxin of var. aizawai, and with polypeptides of 130 and 138 (X 10(3)) Mr from var. thuringiensis. In addition, crystals from var. kurstaki and var. aizawai contained an antigenically related 63 X 10(3) Mr protein that did not cross-react with antisera to the 130 X 10(3) Mr component. Bioassays on Pieris brassicae larvae (Lepidoptera) and Aedes aegypti larvae (Diptera) indicated that the 130 X 10(3) Mr protein of var. kurstaki, and the 138 plus 130 X 10(3) Mr components of var. thuringiensis killed only P. brassicae, while the 130 X 10(3) Mr protein of var. aizawai and the 63 X 10(3) Mr proteins of var. aizawai and var. kurstaki were toxic to both P. brassicae and A. aegypti. Activation of the 130 and 138 (X 10(3)) Mr proteins of the three varieties of B. thuringiensis with insect gut proteases yielded active products of 50-60 (X 10(3)) Mr. Assay of these products on a range of lepidopteran and dipteran cell lines revealed very different toxicity spectra: var. kurstaki killed only one lepidopteran line, var. thuringiensis killed two lepidopteran lines, while var. aizawai was cytolytic to all of the lepidopteran and most of the dipteran cell lines tested, reflecting its broader spectrum in vivo. Thus we have shown that antigenic cross-reaction of B. thuringiensis delta-endotoxins does not necessarily imply a similar toxicity spectrum in vivo or in vitro. PMID- 3027113 TI - Bone marrow transplantation in hematologic malignancies. PMID- 3027114 TI - An enzyme-linked immunosorbent assay for detection of flavivirus antibodies in chicken sera. AB - An enzyme-linked immunosorbent assay (ELISA) was developed to determine the presence of flavivirus antibodies to Murray Valley encephalitis (MVE) and Kunjin viruses in sentinel chicken sera. The development of a quick, reliable assay to detect antibodies to MVE was an essential part of a large-scale surveillance programme to monitor arbovirus activity in Western Australia. This assay was developed for use with alkaline phosphatase conjugated goat anti-mouse IgG using mouse anti-chicken globulin in an intermediate step. There was a significant difference in absorbance values between neutralization test positive and pre-bled sera. However, some sera obtained from sentinel chickens and deemed negative by neutralization demonstrated adsorbance levels above the cutoff level in the ELISA, which reflects the increased sensitivity of this technique. The ELISA test detected antibodies to MVE in chicken sera 7-10 days after infection, whereas these antibodies were only consistently detected by the neutralization test 24 days after infection. Antibodies to both MVE and Kunjin reacted positively with the MVE antigen, but there was little cross-reactivity between this antigen and antibodies to other togaviruses. The main advantages of the ELISA over the neutralization test for detecting antibodies to MVE virus in the sera of sentinel chickens are its greater sensitivity and the speed with which tests can be performed. Results are available within 48 h of receiving specimens and emergency mosquito control measures may then be implemented. PMID- 3027115 TI - Detection of IgA antibody to EBV membrane antigen using Staphylococcus aureus preabsorbed sera is closely associated with nasopharyngeal carcinoma. AB - The presence of IgA antibody to membrane antigen (MA) of Epstein-Barr virus (EBV) was tested in sera from 48 nasopharyngeal carcinoma (NPC) patients, 40 patients with tumors other than NPC and 46 normal individuals. The sera were preabsorbed with Staphylococcus aureus (SPA) (strain no. 1800) prior to their use in the indirect immunofluorescence test. One hundred percent of the NPC patients had the IgA/MA antibody with a GMT of 1:141. In patients with tumors other than NPC or normal individuals, IgA/MA antibodies were not detectable. The IgA/MA antibodies have been demonstrated in 6 NPC patients lacking detectable antibody levels in the indirect immunofluorescence test using nonabsorbed sera. Our data indicate that preabsorbtion of sera with SPA renders the diagnostic test significantly more sensitive for the detection of the nasopharyngeal carcinoma and can be used for trials on the prognosis of patients. PMID- 3027116 TI - Properties of acrylamide gels cross-linked with low concentrations of N,N' diallyltartardiamide. AB - The behavior of proteins on acrylamide gels cross-linked with low concentrations of N,N'-diallyltartardiamide (DATD) has been studied from a theoretical and practical point of view. Modifications in methodology and a technique for calculation of apparent molecular weights (MWs) allowed accurate determinations on gels from 7 up to 15% acrylamide. Comparison of viral protein patterns obtained with DATD- or N,N'-methylenebisacrylamide-cross-linked gels indicated greater resolution by DATD gels, particularly in the higher MW range. The polymerization reaction was studied kinetically and found to be efficient so long as the ratio of DATD to acrylamide was low. High resolution of DATD gels was attributed to the spontaneous formation of a gradient over much of the gel that retarded diffusion of the protein bands. PMID- 3027117 TI - An ELISA for the detection of rhinovirus specific antibody in serum and nasal secretion. AB - An enzyme-linked immunosorbent assay (ELISA) which detects rhinovirus specific antibody in human sera and nasal secretions, has been developed. This sandwich ELISA utilizes a rabbit antirhinovirus hyperimmune serum as the capture antibody and was found to be very sensitive, detecting rhinovirus specific antibody in the serum at dilutions of 1:10(6) and 1:10(3.5) for IgG and IgA immunoglobulins, respectively. Thus, this new assay is 10(2)-10(4) times more sensitive than our standard neutralization test. Furthermore, this increase in sensitivity has enabled us to reliably detect rhinovirus specific immunoglobulins in unconcentrated nasal washings, which are thought to be particularly important for protection against rhinovirus reinfection. A preliminary study of the immune response in human volunteers challenged with rhinovirus using this new ELISA system is presented and further applications and potential of the method are also discussed. PMID- 3027118 TI - Factors affecting the detection of cytomegalovirus in urine by sandwich enzyme immunoassays. AB - Some factors influencing the detection of human cytomegalovirus (HCMV) in urine were investigated employing 2 enzyme-linked immunosorbent assays (ELISAs); one utilised anti-CMV DNA polymerase while the other anti-CMV mouse monoclonals as the detecting antibodies. The use of anti-CMV DNA polymerase was found to be superior in detecting HCMV in both urine and tissue culture fluids than anti-CMV monoclonals. Furthermore, alkaline phosphatase conjugates produced much lower background than did peroxidase conjugates. In reconstruction experiments, the extremes of pH in the urine clearly had an adverse effect on the detection rate of extracellular virus. pH correction of urines to neutrality improved the detection rate considerably. On the other hand, pH correction had little effect on the detection rate of intracellular HCMV in urine, although it was improved when specimens were subjected to repeated cycles of freeze-thawing, ultrasonication, and storage at 4 degrees C. It was concluded that, in addition to the factors investigated which all appear to affect virus detection rate, there may well be additional factors that interfere with CMV detection in the urine by ELISA particularly with intracellular virus. PMID- 3027119 TI - The growth properties and CEA localization of cultured signet ring cell carcinoma transplanted into athymic nude mice. PMID- 3027121 TI - Myosin specific phosphatases isolated from Dictyostelium discoideum. AB - Using native myosin phosphorylated on either the heavy chain or the light chain, we have isolated myosin phosphatases from extracts of vegetative Dictyostelium amoeba. Two phosphatases were resolved by DEAE-cellulose chromatography. One of these phosphatases removed phosphate from heavy chain or light chain at approximately the same rate. The other phosphatase appeared to be much more specific for phosphorylated myosin heavy chain. Although these enzymes removed phosphate from other phosphoprotein substrates such as histone or casein, they did so at a much lower rate. Both enzymes required magnesium for activity, but appeared to be independent of calcium. PMID- 3027122 TI - Lansing poliovirus infection in mice: antibody demonstrable by enzyme-linked immunosorbent assay (ELISA) and immunoprecipitation but not by neutralization. AB - Adult mice infected intracerebrally (i.c) with the Lansing strain of type 2 human poliovirus (HPV2) failed to develop a systemic neutralizing antibody response until 2 months post-infection (p.i). In contrast, an enzyme-linked immunosorbent assay (ELISA) demonstrated an antibody response of IgM and IgG classes beginning at day 1 p.i. with peak levels reached by 5 weeks p.i. This response was slightly greater in paralyzed than in nonparalyzed animals. Immunoprecipitation of poliovirus proteins from cytoplasmic extracts and disrupted purified virion preparations revealed antibodies to three capsid proteins, two capsid precursor proteins, and one nonstructural protein. Finally, neither neutralizing antibody nor definite virus replication was detected after oral, intraperitoneal, or intravenous routes of inoculation. We conclude that the lack of a systemic neutralizing antibody response in mice is probably due to an insufficient amount of infectious virus and consequently viral neutralizing epitopes reaching extraneural lymphoid tissues. PMID- 3027120 TI - Partial purification of two myosin heavy chain kinases from Dictyostelium discoideum. AB - Myosin heavy chain kinase activity was identified in the high speed supernate of lysed Dictyostelium amoebae and was precipitated by 30-50% ammonium sulphate. In low ionic strength buffer, the activity bound tightly to a Cibacron Blue Sepharose column and eluted as a single peak with 1.0 M NaCl. Gel filtration chromatography resolved the kinase into two activities, each of which phosphorylated the tail portion of purified Dictyostelium myosin. One of these activities phosphorylated both serine and threonine residues of the heavy chain, while the other activity only phosphorylated threonine residues. Peptide mapping studies indicated that in vivo and in vitro phosphorylation sites were identical. The heavy chain kinases required Mg2+ for activity but were unaffected by Ca2+ or calmodulin. The heavy chain kinases did not phosphorylate Dictyostelium light chain, and also did not phosphorylate myosins from striated, smooth, or other nonmuscle sources. PMID- 3027123 TI - Effects of regional spinal X-irradiation on demyelinating disease caused by Theiler's virus, mouse hepatitis virus or experimental allergic encephalomyelitis. AB - The effects of X-irradiation on the course of chronic demyelinating disease were examined in mice with experimental allergic encephalitis (EAE), mouse hepatitis virus (MHV) or Theiler's virus (DAV) infection. One month after the induction of EAE or 2-16 months after inoculation of DAV, exposure of the cervical spinal cord to 20 Gy X-rays caused local exacerbation of disease activity but spinal irradiation did not affect MHV-induced demyelination. In EAE, there was a significant increase in the number of inflammatory cells in the irradiated part of the cord. Mice infected with DAV showed locally increased demyelination and axonal degeneration but no change in the titer of infectious virus within the cord. Thus in DAV infection, as in EAE, the exacerbation of disease seemed to be due to vascular or immunological factors rather than viral reactivation. PMID- 3027124 TI - Cardiotonic agent milrinone stimulates resorption in rodent bone organ culture. AB - The cardiotonic agent amrinone inhibits bone resorption in vitro. Milrinone, an amrinone analog, is a more potent cardiotonic agent with lower toxicity. In contrast to amrinone, milrinone stimulated resorption in cultures of neonatal mouse calvaria and fetal rat limb bones. Threshold doses were 0.1 microM in calvaria and 0.1 mM in limb bones; maximal stimulation occurred in calvaria at 0.1 mM. Maximal responses to milrinone and parathyroid hormone were comparable. Milrinone concentrations below 0.1 mM did not affect calvarial cyclic AMP. 0.5 microM indomethacin inhibited milrinone effects in calvaria but usually not in limb bones. 3 nM calcitonin inhibited milrinone-stimulated resorption and there was no escape from this inhibition. Structural homology between milrinone and thyroxine has been reported. We find similarities between milrinone and thyroxine actions on bone, because prostaglandin production was crucial for the effects of both agents in calvaria but not in limb bones, and neither agent exhibited escape from calcitonin inhibition. PMID- 3027125 TI - Protection against the lethal effects of pentobarbital in mice by a benzodiazepine receptor inverse agonist, 6,7-dimethoxy-4-ethyl-3-carbomethoxy beta-carboline. AB - The benzodiazepine receptor inverse agonist 6,7-dimethoxy-4-ethyl-3-carbomethoxy beta-carboline (DMCM) (1.5-15 mg/kg) was administered to mice 5 min after a lethal (LD94) injection of pentobarbital. DMCM (1.5-5 mg/kg) increased short-term (1 h) survival in a dose-dependent fashion, with an optimum survival rate more than five times greater than mice receiving pentobarbital alone. Statistically significant increases in long-term (24 h) survival were also observed after both 5 and 10 mg/kg of DMCM (34 and 33%, respectively) compared with animals receiving pentobarbital alone (6%). Two doses of DMCM (5 and 2.5 mg/kg, respectively) administered 55 min apart produced an even greater increase (58%) in 24-h survival rates. Doses of DMCM that increased 1- and 24-h survival were not lethal when administered alone, and were below the dose that produced convulsions in 50% (CD50) of the animals. The protective effects of DMCM were blocked by pretreatment with the benzodiazepine receptor agonist ethyl-8-fluoro-5,6-dihydro 5-methyl-6-oxo- 4H-imidazo[1,5a][1,4]benzodiazodiazepine-3-carboxylate (Ro 15 1788), which suggests the effects of DMCM are mediated through the benzodiazepine receptor. These findings suggest that DMCM or another benzodiazepine receptor ligand with full inverse agonist qualities could prove effective as an antidote for barbiturate intoxication in man. PMID- 3027126 TI - Human giant cell tumors of bone identification and characterization of cell types. AB - Cells cultured from human giant cell tumors of bone were characterized on the basis of morphological features, proliferative capacity, presence of granulocyte monocyte antigens, receptors for skeletal hormones, and soluble cell products. Three major cell types were identified. One population consisted of mononuclear cells with fibroblastic morphology, which proliferated in culture and most likely represent the neoplastic element of the tumor. Phenotypically they resembled a connective tissue stromal cell. A second population of mononuclear cells lacked receptors for skeletal hormones and did not persist in culture. These cells were likely of monocyte-macrophage lineage. A third population of cells consisted of large multinucleated giant cells. These cells possessed phenotypic features of osteoclasts including receptors for calcitonin. Human giant cell tumors of bone are most likely a neoplasm of connective tissue stromal cells, which have the capacity to recruit and interact with multinucleated giant cells that exhibit phenotypic features of osteoclasts. PMID- 3027127 TI - Effects of atrial natriuretic factor on cyclic guanosine monophosphate and cyclic adenosine monophosphate accumulation in microdissected nephron segments from rats. AB - Atrial natriuretic factor (ANF) (1 microM) markedly increased cyclic guanosine monophosphate (cGMP) content in microdissected glomeruli (35-fold) and in microdissected inner medullary collecting ducts (IMCD) (20-fold). ANF caused little or no increase in cGMP content in other nephron segments. The threshold concentration for increased cGMP accumulation by ANF was 0.1-1 nM in IMCD, which is in the range reported for rat plasma. Sodium nitroprusside (1 mM), which selectively stimulates soluble guanylate cyclase, increased cGMP content in glomeruli but not in IMCD. ANF did not alter cAMP accumulation in the absence or presence of vasopressin (AVP) or parathyroid hormone (PTH) in outer and inner medullary tubule suspensions, or in microdissected proximal convoluted tubules (PCT), medullary thick ascending limbs (MAL) or IMCD. These data are compatible with the hypothesis that cGMP is a second messenger for a physiologic action of ANF in the inner medullary collecting duct. ANF apparently activates membrane bound guanylate cyclase in this segment. PMID- 3027129 TI - Molecular cloning of human synovial cell collagenase and selection of a single gene from genomic DNA. AB - We used a subclone of a rabbit genomic clone for collagenase that cross hybridizes with human synovial cell messenger RNA (mRNA) to identify a human collagenase complementary DNA (cDNA) clone. The human cDNA clone is 2.1 kilobases (kb) and selects a mRNA transcript of approximately the same size from primary cultures of rheumatoid synovial cells that produce collagenase, but no mRNA is selected from control (nonproducing) synovial fibroblasts. Restriction enzyme analysis and DNA sequence data indicate that our cDNA clone is full length and that it is identical to that recently described for human skin fibroblast collagenase. The cDNA clone identified a single collagenase gene of approximately 17 kb from blots of human genomic DNA. The identity of human skin and synovial cell collagenase and the ubiquity of this enzyme and of its substrates, the interstitial collagens types I, II, and III, imply that common mechanisms controlling collagenolysis throughout the human body may be operative in both normal and disease states. PMID- 3027130 TI - Triiodothyronine enhances renal response to aldosterone in the rabbit collecting tubule. AB - Since thyroid hormones and mineralocorticoids were observed to stimulate kidney Na-K-ATPase in similar sites and with similar time courses, this study was initiated to evaluate whether aldosterone is involved in the stimulation of Na-K ATPase observed in collecting tubules 3 h after triiodothyronine (T3) administration to thyroidectomized (TX) rabbits. Results indicate that: Plasma aldosterone level decreased markedly in TX rabbits but was not restored 3 h after T3 injection; Early stimulation of Na-K-ATPase by T3 was abolished when plasma aldosterone level was suppressed by adrenalectomy or when aldosterone effects were blocked by spironolactone; Administration of aldosterone to TX rabbits mimicked the action of T3; Sensitivity of Na-K-ATPase to aldosterone markedly decreased after thyroidectomy. These results demonstrate an interaction between aldosterone and T3 in the control of Na-K-ATPase in the collecting tubule. Triiodothyronine enhances the sensitivity of Na-K-ATPase to aldosterone which, in turn, produces a stimulatory action despite the decreased plasma level observed during hypothyroidism. PMID- 3027131 TI - Localization of the gene for coagulation factor XIII a-chain to chromosome 6 and identification of sites of synthesis. AB - Factor XIII, the clotting factor essential for covalent stabilization of the fibrin clot, is a heterodimer consisting of a2 and b2 subunits, with catalytic function residing in the a-chain. In order to address questions regarding sites of synthesis and chromosomal localization of the Factor XIII a-chain, cDNA was cloned from a lambda gt11 human placental cDNA library. Nucleotide and amino acid sequences were determined from the cDNA. Amino acid sequencing of purified platelet Factor XIII a-chains confirmed the authenticity of the lambda gt11 clone. The gene for Factor XIII a-chain was mapped uniquely to chromosome 6. Northern blot analysis of human placental and U937 (monocytelike) cell poly (A)+ mRNA showed a single approximately 4.0-kb message for the Factor XIII a-chain. These results provide conclusive evidence that the a-chain is synthesized by placenta and monocyte cell lines. PMID- 3027128 TI - Role of activation of protein kinase C in the stimulation of colonic epithelial proliferation and reactive oxygen formation by bile acids. AB - Deoxycholate (DOC), chenodeoxycholate, 12-O-tetradecanoyl phorbol-13-acetate (TPA), or 1-oleoyl-2-acetyl-glycerol (OAG) activated colonic epithelial protein kinase C as reflected by translocation from the soluble to the particulate cell fraction. Activation of protein kinase C was correlated with stimulation of enhanced proliferative activity of colonic mucosa and reactive oxygen production. TPA and OAG, but not DOC, directly activated soluble protein kinase C in vitro. However, DOC rapidly increased labeled inositol phosphate and diacylglycerol accumulation in colonic epithelial cells. Retinoic acid inhibited protein kinase C activity and suppressed DOC-, TPA-, and OAG-induced increases in reactive oxygen production. The results support a role for protein kinase C in the stimulation of colonic epithelial proliferative activity and reactive oxygen production induced by bile acids, TPA and OAG. In contrast to TPA and OAG, which activate protein kinase C directly, bile acids appear to activate protein kinase C indirectly by increasing the diacylglycerol content of colonic epithelium. PMID- 3027133 TI - Hemoglobin potentiates central nervous system damage. AB - Iron and iron compounds--including mammalian hemoglobins--catalyze hydroxyl radical production and lipid peroxidation. To determine whether hemoglobin mediated lipid peroxidation might be important in hemorrhagic injuries to the central nervous system (CNS), we studied the effects of purified hemoglobin on CNS homogenates and injected hemoglobin into the spinal cords of anesthetized cats. Hemoglobin markedly inhibits Na/K ATPase activity in CNS homogenates and spinal cords of living cats. Hemoglobin also catalyzes substantial peroxidation of CNS lipids. Importantly, the potent iron chelator, desferrioxamine, blocks these adverse effects of hemoglobin, both in vitro and in vivo. Because desferrioxamine is not known to interact with heme iron, these results indicate that free iron, derived from hemoglobin, is the proximate toxic species. Overall, our data suggest that hemoglobin, released from red cells after trauma, can promote tissue injury through iron-dependent mechanisms. Suppression of this damage by desferrioxamine suggests a rational therapeutic approach to management of trauma-induced CNS injury. PMID- 3027132 TI - Distribution of glucose transporter messenger RNA transcripts in tissues of rat and man. AB - We used the complementary DNA for the human hepatoma Hep G2 glucose transporter to determine the distribution of glucose transporter messenger RNA (mRNA) in rat and human tissues. Under stringent hybridization conditions, a single 2.8 kilobase (kb) transcript is seen in all rat and human tissues examined. The mRNA is most abundant in brain, and is especially enriched in the brain microvascular fraction. The mRNA abundance in rat muscle and fat is 5% that in brain. Rat liver (both adult and fetal) and human liver have very little 2.8-kb mRNA, but it is abundant in cultured human fibroblasts and EB virus-transformed lymphoblasts. The same size mRNA is present in leg muscle of two type II diabetic patients. A very homologous glucose transporter mRNA is expressed in both insulin-sensitive and insensitive tissues of rat and man. Hepatocytes, which have abundant glucose transport, may express a homologous but nonidentical glucose transporter. PMID- 3027134 TI - Detection of infectious pancreatic necrosis virus using an inhibition enzyme linked immunosorbent assay. AB - An inhibition enzyme-linked immunosorbent assay was used to detect infectious pancreatic necrosis (IPN) virus. In this assay the presence of virus was determined by measuring the decrease in titer of a known antiserum after incubation with a sample suspected to contain virus. The titer of the antiserum was measured with an indirect enzyme-linked assay. Compared to the double antibody sandwich method this assay required fewer reagents (only one anti-IPN serum was required). This assay was also sensitive enough to detect virus at levels of 1 X 10(2) TCID 50/ml. of purified virus and was able to detect virus in samples obtained in the field. PMID- 3027135 TI - Immunohistological studies of lymphoproliferative lesions in a fatal case of Epstein-Barr virus infection. AB - A fatal case of infectious mononucleosis occurred in a young adult. Abnormal serological features were noted in his mother, although there was no other family history suggesting an inherited defect of immune response to Epstein-Barr virus (EBV). The cellular infiltrate observed in tissues obtained at necropsy was analysed with a range of specific monoclonal and polyclonal antibodies. Polyclonal plasmacytoid B cell proliferation had occurred in many tissues. These cells were positive for EBV nuclear antigen, but viral particles were not seen on ultrastructural examination, and the virus was not isolated, suggesting a non permissive infection. PMID- 3027136 TI - Sodium valproate-induced analgesia: possible role of the GABA-ergic system in pain mechanism. PMID- 3027137 TI - Synaptic connections of cholecystokinin-immunoreactive neurons and terminals in the rat fascia dentata: a combined light and electron microscopic study. AB - We report here on the fine structure and synaptic connections of neurons and axon terminals in the rat fascia dentata displaying immunoreactivity to antibodies against cholecystokinin octapeptide (CCK). In the fascia dentata and hilar region, CCK-immunoreactivity was confined to nonpyramidal neurons that were similar in appearance to basket cells known to use gamma-aminobutyric acid (GABA) as neurotransmitter. These neurons exhibited dense accumulations of endoplasmic reticulum and infolded nuclei, and established asymmetric and symmetric synaptic contacts with presynaptic terminals. Among those terminals that formed asymmetric synaptic contacts, giant mossy fiber boutons arising from granule cell axons were identified. Cholecystokinin-immunoreactive terminals established symmetric synaptic contacts on the cell bodies and dendrites of granule cells. Similar contacts were formed on nonimmunoreactive hilar neurons. Some of these hilar cells were identified as commissural neurons by retrograde filling with horseradish peroxidase (HRP) following injection of the tracer into the contralateral fascia dentata. Synaptic contacts were rarely observed between immunolabeled pre- and postsynaptic elements. The results are discussed with regard to inhibitory processes in the fascia dentata since other studies have shown that CCK is coexistent with GABA in hippocampal nonpyramidal neurons. PMID- 3027138 TI - Monocyte and neutrophil chemotaxis in psoriasis. Relation to the clinical status and the type of psoriasis. AB - Chemotactic activity of purified monocytes and polymorphonuclear leukocytes was studied in fifty patients with psoriasis vulgaris and in forty-five healthy individuals by an objective in vitro assay with the use of a 51Cr-labeling technic. Both monocytes and polymorphonuclear leukocytes showed a statistically significant increase in chemotactic response, which was positively correlated with disease activity but not with the extent of the cutaneous lesions. The chemotactic activity of monocytes correlated with that of polymorphonuclear leukocytes in the same patients. Exacerbation of psoriasis was preceded by a rapid increase of polymorphonuclear leukocyte chemotaxis, and a decline of chemotaxis occurred during clinical improvement. The psoriatic leukocytes were 22% more sensitive than normal leukocytes to leukotriene B4 than to N-formyl methionyl-leucyl-phenylalanine. Psoriatic plasma showed chemotaxis-enhancing properties, but only in patients with widespread cutaneous lesions. Additionally, monocyte and polymorphonuclear leukocyte chemotaxis was studied in twenty patients with pustular psoriasis and in fifteen patients with psoriatic arthritis. The chemotactic profiles in pustular psoriasis were different from those in psoriasis vulgaris. Patients with pustular psoriasis had significantly higher polymorphonuclear leukocyte chemotaxis than patients with psoriasis vulgaris, but the chemotactic activity of monocytes was normal. The presence of seronegative arthritis had no influence on chemotactic activity of psoriatic leukocytes. PMID- 3027140 TI - Coexistence of angiolymphoid hyperplasia with eosinophilia and pulmonary neoplasia. PMID- 3027139 TI - Extramammary Paget's cells: further evidence of sweat gland derivation. AB - Tissue from a 66-year-old male patient with extramammary Paget's disease of the right side of the scrotum and the right groin was studied for the presence of several antigens with the immunoperoxidase technic. Adenokeratin (cytokeratin No. 18) and carcinoembryonic antigens were positive in Paget's cells, whereas squamokeratin (cytokeratin No. 10) was expressed only in normal epidermal cells. Langerhans cells were decreased in the region of the tumor. Many transferrin receptors were present on the tumor cells, indicating a high cellular proliferation rate. Enzyme histochemical studies of the extramammary Paget's cells showed positive reactions for several enzymes typical of sweat glands, except for leucine aminopeptidase, which was negative. A comparison with three other cases showed that these enzyme reactions varied greatly from case to case. Both immunohistochemical and enzyme histochemical findings provide further evidence that extramammary Paget's cells are related to sweat gland epithelial cells, with variable expression of cellular characteristics. PMID- 3027142 TI - CT diagnosis of massive hemorrhage from hepatocellular carcinoma. AB - Massive hemorrhage from rupture of hepatocellular carcinoma is uncommon. We report our experience in two cases diagnosed by CT. PMID- 3027141 TI - Fast spin echo imaging with suspended respiration: gadolinium enhanced MR imaging of liver tumors. AB - Magnetic resonance (MR) imaging of 43 patients with hepatic tumor was performed during suspended respiration using a fast scan spin echo (SE) technique (SE 200/40) with a single excitation. The resulting images were superior in terms of image quality to conventional ones. Due to the lack of soft tissue contrast, 38 patients received Gd-diethylenetriaminepentaacetic acid (DTPA) at 0.05 mmol/kg and serial scanning (CE-MR) was repeated. Twelve of 14 hepatomas showed isointensity or slightly reduced intensity compared with the liver in unenhanced MR. All metastases (nine patients) showed low signal intensity that was statistically significant (p less than 0.001) in differentiating between hepatomas and metastatic liver tumors. With contrast enhanced MR, both hepatomas and metastases showed changes that cannot be further classified until more cases have been examined. In all 12 cavernous hemangioma cases, Gd-DTPA pooling was observed with extremely high contrast, which was a pathognomonic sign. In fact, four cavernous hemangiomas in two patients with a diameter of 1.0 cm were successfully imaged. PMID- 3027143 TI - CT findings in splenic hemangiomas in the Klippel-Trenaunay-Weber syndrome. AB - The Klippel-Trenaunay-Weber syndrome consists of cutaneous port wine hemangiomas, superficial venous varicosities, and soft tissue and bony hypertrophy of an extremity. We describe three patients with this syndrome with hypodense lesions in the spleen which in one case became isodense after bolus contrast CT. In one case ultrasound found numerous echopenic and echogenic masses. These lesions are presumed to represent multiple hemangiomata. PMID- 3027144 TI - Rumen liquid dilution rate in dairy cows fed once daily: effects of diet and sodium bicarbonate supplementation. AB - Three ruminally cannulated Holstein cows fed total mixed diets of hay crop silage and concentrate (30:70, 50:50, 70:30, 100:0) were used to determine liquid volume ruminal and dilution rate with polyethylene glycol as a marker. Dosing 5 h prefeeding produced nonlinear marker dilution curves that were divided into prefeeding, eating, and resting phases, whereas dosing 2 h postfeeding produced linear dilution curves. Ruminal liquid volume calculated from either dosing method overestimated liquid volume compared to manual estimates. Mean liquid dilution rates did not differ between methods of dosing, and differences between diets were closely related to dry matter intake. Supplementation with 0, 68, or 114 g/day sodium bicarbonate did not affect ruminal liquid volume or dilution rates. Across diet and buffer treatments, liquid dilution rate during eating (23 to 32%/h) was greater than mean, prefeeding, or resting dilution rates (10 to 21%/h). Correct interpretation of ruminal volume and dilution data requires full details of dosing method, dosing time in relation to feeding, and sampling times. PMID- 3027146 TI - Effects of sodium bicarbonate with three ratios of hay crop silage to concentrate for dairy cows. AB - Three ruminally cannulated Holstein cows were fed total mixed diets of hay crop silage and concentrate (30:70, 50:50, 70:30, 100:0) to evaluate effects of sodium bicarbonate supplements equivalent to 0, .4, and .7% of total ration dry matter (0, 68, and 114 g/d). Yields of milk, fat-corrected milk, fat, protein, and solids-not-fat, percentages of milk protein and solids-not-fat, and efficiency of production of fat-corrected milk declined with decreasing concentrate proportion. Buffer supplementation reduced milk fat percentage and milk yield was greater with 68 g/d sodium bicarbonate than with 114 g/d. Digestibilities of dry matter, organic matter, gross energy, cell solubles, and crude protein declined with decreasing proportion of concentrate while cellulose digestibility increased linearly. The proportion of dietary nitrogen transferred to milk decreased linearly with decreasing proportion of concentrate and sodium bicarbonate increased this transfer with the 70% concentrate diet. Sodium bicarbonate increased ruminal pH and acetate proportion while decreasing ammonia concentration. Acetate:propionate ratio was decreased by sodium bicarbonate addition to the 70% concentrate diet. High concentrate diets with hay crop silage may require higher amounts of buffers to influence production. PMID- 3027145 TI - Effect of prepartum energy, body condition, and sodium bicarbonate on production of cows in early lactation. AB - In trial 1, the effects of dietary energy (102, 131 or 162% of requirement) in the dry period and of sodium bicarbonate (0 or .75% of diet dry matter) in early lactation were assessed with 31 cows in a 3 X 2 factorial arrangement of treatments. Body condition and weight increased linearly with prepartum energy. Dry matter intake and milk yield were similar across treatments through 12 wk postpartum. Sodium bicarbonate increased milk fat content only in the 131% group, an effect apparently related to greater mobilization of fat in that group. In trial 2, energy treatments imposed in late lactation (145 to 55 d prepartum) and in the dry period (55 to 0 d) were 1) cows fed to requirement in both periods, 2) cows overfed in the first and underfed in the second period, 3) cows fed to requirement in the first and overfed in the second period, and 4) cows overfed in both periods. Cows in treatments 1 and 2 (normal) calved in a thinner state than those in 3 or 4 (fat). In the first 12 wk postpartum, intake did not differ, but cows in groups 3 and 4 produced more milk. Sodium bicarbonate imposed factorially postpartum increased milk fat content. Overconsumption of energy prepartum did not impair production when high energy total mixed rations were fed postpartum. PMID- 3027147 TI - Physical parameters of fiber affecting passage from the rumen. AB - Our objective was to review the factors that affect fiber passage from the rumen. Rumen residence time and passage from the rumen are important in control of intake, digestibility, protein metabolism, and protein escape. Physical factors associated with particle size and particle specific gravity affect passage from the rumen. Although particles of 5 cm may pass through the reticulo-omasal orifice, most particles leaving the rumen are smaller than 1 mm. Polypropylene ribbon cut into 7-cm lengths, introduced into the rumen, markedly reduced intake and were ruminated to fine particle size before being passed. Materials with specific gravity less than 1.0 are ruminated extensively and are passed slowly. As specific gravity of plastic particles increase, rumination of particles is decreased and passage is increased. Particles with specific gravities between 1.17 and 1.42 pass most rapidly. Increasing specific gravities further results in a decline in passage and a further lowering of rumination of particles. Natural hay and fresh forages are hydrated in the rumen, which causes functional specific gravity to increase. Some factors that affect rate of change of functional specific gravity of forages have been investigated. Small particle size, autoclaved rumen fluid, buffer solutions, and specific salts increased the rate of change of functional specific gravity of particles. Understanding of these factors may enable us to make better decisions in managing ruminant productions systems. PMID- 3027148 TI - Chemical factors involved in ruminal fiber digestion. AB - In the United States, cattle are commonly fed diets containing cereal grains. The presence of starch and sugars reduces fiber digestion, which may in turn depress intake. In this paper, chemical constraints that may be responsible for the decrease in fiber digestion are explored. A major factor appears to be rumen pH. Moderate depression in pH, to approximately 6.0, results in a small decrease in fiber digestion, but numbers of fibrolytic organisms are usually not affected. Further decreases to 5.5 or 5.0 result in depressed growth rates and decreased fibrolytic microbes, and fiber digestion may be completely inhibited. Proliferation of organisms on readily fermentable carbohydrates may increase the need for total nitrogen as both ammonia and amino acids. The value of amino acids to cellulolytic organisms appears to be primarily as sources of isobutyric, isovaleric, and 2-methylbutyric acids. This reinforces the need to establish dietary requirements for nonprotein nitrogen, degradable protein, and isoacids. Other factors affecting fiber digestion, such as inhibition of cellulytic enzymes and plant concentrations of lignins and phenyl propanoids, are also discussed. PMID- 3027149 TI - Use of 1 alpha-hydroxyvitamin D3 to prevent bovine parturient paresis. VI. Concentrations of vitamin D metabolites and vitamin D3 equivalence in milk. AB - Concentration of vitamin D metabolites was determined in the milk of control and 1 alpha-hydroxyvitamin D3-injected (700 micrograms) cows that calved 36 to 43 h after treatment. Milk samples were taken 60 h after calving. Concentrations of vitamin D, 25-hydroxyvitamin D, 24,25-dihydroxyvitamin D, and 1,25 dihydroxyvitamin D in milk of the control cows were 372 +/- 24, 264 +/- 68, 68 +/ 26, and 21 +/- 3 ng/L, respectively. Concentrations of vitamin D metabolites in the milk of the treated cows did not differ significantly from those of controls. Concentration of 1 alpha-hydroxyvitamin D3 in milk of treated cows was less than 20 ng/L. In a second experiment, cows were injected twice, at 72-h intervals, with 350 micrograms 1 alpha-hydroxyvitamin D3. Milk was taken 60 h after parturition from cows that calved 37 to 60 h after the second injection. The vitamin D3 equivalence of the milk was 40 +/- 3 IU/L. Results indicate that injection of 700 micrograms 1 alpha-hydroxyvitamin D3 did not affect the concentration of vitamin D metabolites or the vitamin D3 equivalence of milk taken 60 h after calving. PMID- 3027151 TI - Control of stable flies (Diptera: Muscidae) with a unique nitrogen fertilizer, calcium cyanamide. PMID- 3027150 TI - Effects of dietary calcium, vitamin D3, and corn supplementation on growth performance and mineral metabolism in young goats fed whole milk diets. AB - Twenty-four male goats, approximately 2 to 4 wk of age, were allotted to four dietary treatments in a 2 X 2 factorial design and were fed a basal milk diet at 12.5% body weight for 20 wk. Vitamin D3 was added to the milk in two different amounts with and without supplemental CaCO3. At the end of wk 7, corn was added to all diets at 1% body weight. Over 20 wk, average daily gain was unaltered by addition of Ca or vitamin D3 to the diet. When corn was added to the diet, gains increased from 48 to 180 g/d. Plasma Ca concentrations were not affected by dietary treatment. Supplemental Ca decreased plasma Mg concentrations. Corn supplementation curtailed a depression in plasma Mg and seemed to prevent a whole milk-induced hypomagnesemia. Fecal excretion of all minerals measured was increased in goats fed supplemental Ca. Dietary Ca increased urinary Ca but decreased urinary Mg. Percentage of apparent absorption of Ca, Mg, and total ash was lower in Ca-supplemented goats, as was apparent retention of Ca and Mg. The physiological responses reported suggest the goat as a potential research model for mineral metabolism studies in other ruminants. PMID- 3027152 TI - Elevated mononuclear leukocyte phosphodiesterase in allergic dogs with and without airway hyperresponsiveness. AB - To investigate if mononuclear leukocyte beta-adrenergic hyporesponsiveness of Basenji greyhound (BG) dogs is associated with atopy or nonspecific airway hyperresponsiveness, we examined the relationship between mononuclear leukocyte cAMP phosphodiesterase levels, airway responsiveness to methacholine, and intradermal allergen responses in 17 BG dogs, five unrelated purebred Basenjis, and five greyhounds. BG dogs were hyperresponsive to aerosols of methacholine compared to Basenjis and greyhounds. Both BG dogs and Basenjis were allergic and had increased leukocyte cAMP phosphodiesterase activity compared to greyhounds. We concluded that the leukocyte abnormality is not associated with airway hyperresponsiveness. The leukocyte abnormality is either associated with the allergic state, with some hereditary trait that BG dogs acquired from the Basenji ancestry, or the leukocyte abnormality is necessary but not sufficient for the development of airway hyperresponsiveness. PMID- 3027153 TI - The preventive effect and duration of action of nedocromil sodium and cromolyn sodium on exercise-induced asthma (EIA) in adults. AB - Nedocromil sodium, 4 mg, from a metered-dose inhaler, cromolyn sodium, (cr) 20 mg, from a Spinhaler, and placebo (pl) were compared in their efficacy and duration of action in preventing exercise-induced asthma. Twelve patients with asthma performed treadmill exercise tests 20 minutes, 2 hours, and 4 hours after a single dose of drug in a double-blind, crossover trial. Both active drugs were significantly better than pl at 20 minutes. Two hours after drug administration, only cr was significantly different from pl. The direct comparison between nedocromil and cr demonstrated no significant difference on FEV1, and the only significant difference was with forced expiratory flow between 25% and 75% of vital capacity at 2 hours. It is concluded that at these clinically recommended doses, both drugs are equally effective in preventing exercise-induced asthma with cr possibly having a somewhat longer duration of action. PMID- 3027154 TI - Imipramine prevention of carbon tetrachloride-induced liver necrosis at late states of the intoxication process. AB - Imipramine administration (50 mg kg-1, i.p.) to Sprague-Dawley male rats (240-290 g) 6 or 10 h after CCl4 (1 ml kg-1, i.p.) partially prevents liver necrosis induced by the hepatotoxin. When imipramine is given 30 min before CCl4, it inhibits in part the CCl4-induced lipid peroxidation and the covalent interactions of reactive metabolites with microsomal lipids or proteins and partially prevents CCl4-induced cytochrome P-450 destruction, but not glucose 6 phosphatase activity depression. Imipramine administration prior to CCl4 does not modify levels of the hepatotoxin reaching the liver or the body temperature of CCl4 treated animals. Early preventive effects of imipramine on cytochrome P-450, might be attributed to inhibition of covalent interactions of reactive metabolites. The hypothesis that imipramine exerted late preventive effects by interfering with calcium deleterious effects or by modulation of protein and phospholipid synthesis or degradation is analyzed. PMID- 3027156 TI - New type of linker useful for cloning experiments. AB - A new class of linker oligodeoxynucleotide sequences is described which allows the original sequence of a foreign DNA to be restored after cloning. In the standard linker approach the terminal base pairs of the linker-sequences are irreversibly attached to the cloned DNA, thus altering the genetic information. The use of the new type of linker is demonstrated by the transformation of the unique Pvu II site in pBR322 into Bam HI site using the ISO ('in-site-out') linker d(GATCCGGATC). Possible further applications of these linker sequences are described. PMID- 3027155 TI - The effect of cadmium on chemical- and viral-induced tumor production in mice. AB - Female Swiss mice were exposed to cadmium in the drinking water at concentrations ranging from 0 to 50 ppm for 105 or 280 day time periods. In the 105 day study, the effect of cadmium on urethan-induced pulmonary adenoma formation was evaluated. Urethan-induced sleeping times observed following i.p. injection of urethan after 3 weeks of cadmium exposure were not affected by cadmium indicating that chronic cadmium exposure did not alter the elimination of urethan. Pulmonary adenoma formation which was evaluated 84 days later was not affected by cadmium. The size and number of tumors remained unchanged. This suggests that the immunosuppressive actions of cadmium do not influence urethan-induced adenoma formation. In the 280-day study, the effects of cadmium on the incidence of spontaneous murine lymphocytic leukemia was evaluated. Mortality from the leukemia virus was greater in the cadmium-exposed mice. Mice exposed to 10 or 50 ppm cadmium experienced 33% more deaths from the virus. The average time till death was unaffected. It appears that the immunosuppressive effects of cadmium impair immunosurveillance mechanisms that control expression of the murine lymphocytic leukemia virus. PMID- 3027157 TI - [Triploidy]. AB - Using their own experience which they described in 1984 and reviewing as well the literature of the condition since that time has led the authors to suggest that triploidy should be considered as a pathological entity with three clinical forms: early abortion, mid-trimester termination and the birth of triploidy fetuses either alive or dead. They discuss the clinical features, the ultrasound appearances, the pathological and anatomical details and the cytogenetics of triploidy. They differentiate this condition from trophoblast disease and from the central hydropic conditions which occur with normal caryotypes. They emphasize that there is a difference in the way classical molar pregnancy evolves and repeat again that the term "embryonic mole" should be avoided. PMID- 3027158 TI - [Uterine cervix dysplasias. New therapeutic strategy based on virological data]. AB - There is no consistency in the treatment of dysplasias of the cervix of the uterus. It varies from team to team and there is no criterion that points with any certainty to how the lesion will develop--whether it will become worse, whether it will persist as it is or whether it will regress. For the first time there is an objective factor, namely typing of the papillomavirus by molecular hybridisation. The presence of HPV 16, 18 or 33 is prognostically bad. Active treatment should be instituted. In the other cases "armed surveillance" or destroying the lesions is sufficient. This is particularly important in young women who wish to have a family. PMID- 3027159 TI - Malignant fibrous histiocytoma involving a digit. AB - A case of malignant fibrous histiocytoma (MFH) involving phalanges of a single digit is reported. MFH of the bones of the hand has not previously been reported. Ten years after wide local excision, the patient remains clinically free of disease. PMID- 3027160 TI - Sodium bicarbonate: burst stomachs and high sodium. PMID- 3027161 TI - Nonfunctioning islet cell tumor presenting as gastrointestinal bleeding. PMID- 3027162 TI - Epidemiological investigations of Japanese encephalitis outbreak in Gorakhpur and Deoria districts of Uttar Pradesh 1985. PMID- 3027163 TI - Effect of raised type-3 poliovaccine dose on virus excretion. AB - Excretion of attenuated polioviruses after controlled type 3 poliovaccine administration was studied in the district city of Cheb. The usual type 3 dose, 10(5) TCD50, was doubled in some vaccinees. A total of 972 stool samples collected from primary vaccinees and their family members were examined; three samplings were carried out in most of the subjects. At first sampling, 9 weeks after administration of type 1 + 2 mixture and 1 day before administration of type 3, all poliovirus types were isolated from primary vaccinees, but family members were negative. At second sampling, 14 days after type 3 administration, primary vaccinees displayed suppression of type 1 and 2 replication and a not very pronounced increase in type 3 excretion. Family contacts yielded all types, the least frequent being type 1. At sampling three, 5 months after type 3 administration, the not very clear-cut difference (at 2nd sampling) between vaccinees given the two different type 3 doses was completely obliterated; type 1 was not isolated; and type 2 isolations increased above the 2nd-sampling value. The number of type 1 and 3 isolations decreased in family contacts, in contrast to type 2, where an increase occurred. Excretion of types 1 and 2 at the time of type 3 administration did not demonstrably influence the subsequent excretion of type 3. Mothers were contact infected at a slightly higher rate than other family members. Only a part of excretors among family members of vaccinees excreted virus of the same type as the vaccinees (simultaneously or subsequently). PMID- 3027164 TI - Clustered cases of paravaccinia in milkers. AB - Clustered cases of a disease in men and cows firstly diagnosed as cowpox has been described. Clinical manifestation, epidemiology and laboratory diagnostics are presented. Virological and serological investigations of the specimens taken from sick persons and animals proved paravaccinia virus (genus Parapoxvirus) to be etiological agent of the illness. Cowpox virus (genus Orthopoxvirus) greatly differs from the latter by phenotypical markers. The disease in humans is called Milkers' modules, in cows--pseudocowpox. The disease is mostly benign and this is a reason why it does not attract sufficient attention of health personnel. Nevertheless this infection can cause a certain economical loss. PMID- 3027165 TI - Acquired immunity and allergy of cells cultured in vitro. AB - It is in the reinoculated cell line FL that the sensitivity of cells under the action of a small dose of diphtheria toxin (DT) changes to the toxin itself as well as to the Coxsackie enterovirus. (B-5). After surviving the infection and getting again "healthy", the cells have acquired simultaneously an elevated sensibility to DT and reduced susceptibility to B-5 transmissible even in long passages. These acquired properties are a transient phenomenon, after a certain number of passages the culture returns to initial stage with all properties inherent in the initial line of cells: with regard to DT in 10 months, to coxsackie virus in 7 months. The stage of elevated sensibility to DT and reduced susceptibility to B-5, however, may be not only renewed but also considerably prolonged and even strengthened by repeated contact of cells with a small dose of DT. The small dose of DT is the first delusion of the toxin not provoking any visible morphological changes in the monolayer. The experimental data obtained render it possible to presume that application of reinoculated cell cultures offers a promising method for studying manifestations of laws governing the third factor of immunity and allergy at the level of cells. PMID- 3027166 TI - Phorbol esters and dioctanoylglycerol block anti-IgM-stimulated phosphoinositide hydrolysis in the murine B cell lymphoma WEHI-231. AB - Cross-linking of membrane IgM (mIgM) on both normal resting B cells and on the murine B cell lymphoma WEHI-231 activates the phosphoinositide signal transduction pathway. The initial event in this pathway is the hydrolysis of phosphatidylinositol 4,5-bisphosphate (PtdInsP2), which results in the generation of two second-messengers: inositol trisphosphate (InsP3), which can cause the release of Ca2+ from intracellular stores, and diacylglycerol (DG), which activates protein kinase C. In examining the effects of exogenous activation of protein kinase C on WEHI-231 cells, we found that phorbol esters blocked some of the biologic effects of anti-IgM on WEHI-231 cells. The mechanism of this effect was investigated. Phorbol ester treatment of WEHI-231 cells blocked the ability of anti-IgM to stimulate production of inositol phosphates and accumulation of phosphatidic acid, the phosphorylated product of DG. Phorbol esters also blocked the ability of anti-IgM to cause an increase in intracellular Ca2+. Thus, it is clear that phorbol esters block anti-IgM-stimulated PtdInsP2 hydrolysis in WEHI 231 cells. In addition, a synthetic DG, dioctanoylglycerol (diC8), also blocked anti-IgM-stimulated inositol phosphate production and the anti-IgM-stimulated rise in cytoplasmic Ca2+. The ability of phorbol esters and diC8 to block mIgM mediated signaling may reflect a feedback inhibition mechanism by which activated protein kinase C limits the magnitude and duration of receptor signaling. PMID- 3027167 TI - Accumulation and chemotaxis of natural killer/large granular lymphocytes at sites of virus replication. AB - A model for monitoring the accumulation of natural killer cell/large granular lymphocytes (NK/LGL) at a site of virus replication was studied by using mice infected i.p. with either lymphocytic choriomeningitis virus (LCMV), murine cytomegalovirus (MCMV), mouse hepatitis virus (MHV), Pichinde virus, or vaccinia virus. An i.p. but not i.v. infection resulted in a localized increase in NK/LGL cell number (a fourfold to greater than 20-fold increase) and augmentation (a 10- to 20-fold increase) of NK cell activity associated with virus-induced peritoneal exudate cell (PEC) populations. An increase in NK/LGL cell number was detected as early as 12 hr postinfection (p.i.) and peaked at 3 days p.i. with MHV. The initial LGL recruited into the peritoneal cavity at 1 to 3 days p.i. were nonadherent to plastic and were demonstrated to have an NK cell phenotype: asialo GM1+, Thy-1.2 +/-, Lyt-2.2-, and J11d-. The peak number of LGL appeared at 7 days after infection with the NK cell-resistant virus, LCMV. This LGL population had been previously demonstrated to contain cytotoxic T lymphocyte/LGL (CTL/LGL) as well as NK/LGL. During an MHV infection the number of LGL decreased between days 3 and 7 p.i., suggesting that the second wave of CTL/LGL was absent. These findings may explain the absence of a good MHV-CTL model. Virus-induced, activated NK/LGL responded to chemotactic signals by migrating in a unidirectional manner across two 5-microns pore size polycarbonate filters during 7 hr in vitro chemotaxis assays. Wash-out fluid obtained from the peritoneal cavity contained chemotactic activity for NK/LGL as well as for other cell types. We conclude that production and/or release of chemotactic factors at sites of virus replication are at least partially responsible for the accumulation of NK/LGL at these sites. PMID- 3027168 TI - Sera from HTLV-III/LAV antibody-positive individuals mediate antibody-dependent cellular cytotoxicity against HTLV-III/LAV-infected T cells. AB - The causative agent of the acquired immunodeficiency syndrome (AIDS) has been shown to be a human retrovirus called human T lymphotropic virus (HTLV)-III or lymphadenopathy-associated virus (LAV). The nature of the protective immune response against this virus is currently unknown. We report here results using an antibody-dependent cellular cytotoxicity (ADCC) assay which has been developed for measuring a specific immune response against HTLV-III/LAV. Forty-four sera were examined for their ability to mediate ADCC against HTLV-III/LAV-infected T cells. Sera from healthy HTLV-III/LAV seropositive individuals in the presence of mononuclear cells from healthy HTLV-III/LAV seronegative donors exhibited significantly higher levels of ADCC activity compared to sera from patients with AIDS. Western blot analysis of serum samples indicated that antibody reactivity with the p24 protein of HTLV-III/LAV correlated with higher levels of ADCC activity than did reactivity with Gp120/160. The observation that sera from healthy HTLV-III/LAV seropositive individuals mediated higher levels of ADCC activity than did sera obtained from subjects with AIDS suggests that ADCC may represent a protective immune response to infection with HTLV-III/LAV. PMID- 3027169 TI - Inhibition of human B cell responsiveness by prostaglandin E2. AB - The capacity of prostaglandin E2 (PCE2) to modulate human peripheral blood B cell proliferation and the generation of immunoglobulin-secreting cells (ISC) stimulated by Cowan 1 strain Staphylococcus aureus and mitogen-stimulated T cell supernatant was examined. PGE2 significantly inhibited both responses, whereas PGF2 alpha had no inhibitory effect. Responses of highly purified B cells obtained from spleen, lymph node, and tonsil were also inhibited. In addition PGE2 suppressed B cell responses supported by recombinant interleukin 2 rather than T cell supernatant. PGE2-mediated inhibition was mimicked by forskolin, a direct activator of adenylate cyclase. Kinetic studies indicated that PGE2 inhibited B cell responses by a progressively greater increment as cultures were prolonged. Evaluation by flow cytometry after staining with acridine orange or mithramycin indicated that PGE2 had no effect on initial B cell entry into the G1 phase of the cell cycle, passage through G1, and entry into S, G2, and M. Rather, PGE2 inhibited responses of postdivisional daughter cells. PGE2 inhibited responses in cultures stimulated by the calcium ionophore ionomycin and T cell supernatant but had minimal effects in cultures stimulated by the combination of ionomycin and phorbol myristate acetate. Moreover, phorbol myristate acetate reversed PGE2-mediated inhibition of proliferation stimulated by S. aureus or S. aureus + T cell supernatant. These results indicate that PGE2 suppresses the continued growth and differentiation of human B cells, although it has no effect on initial B cell activation and suggest that PGE2 may play a role in regulating human B cell responses in vivo. PMID- 3027171 TI - Behavior of aldehyde moieties involved in the activation of suppressor cells by sodium periodate. AB - The treatment of mouse spleen cells with periodate at the optimal mitogenic concentration (1 mM) induces the activation of suppressor cells of the in vitro antibody response and leads to the formation of aldehydes on the carbohydrate termini of the surface sialoglycoconjugates. These aldehyde moieties are found on the C8 (N-AN 8) and the C7 (N-AN 7) derivatives of sialic acid. Immediate borohydride reduction prevents the activation of the suppressor cells. Data from this work show that borohydride reduction must be performed within the first 6 hr to prevent the generation of suppressor cells; 18 hr after the initial periodate oxidation, borohydride treatment did not reverse the in vitro suppressive activity of periodate-treated cells. The kinetics of the disappearance of aldehydes from the cell surface were studied by using [3H]borohydride labeling and chromatographic analysis of sialic acid derivatives. About 70 to 80% of the aldehyde moieties were found to be present 6 hr after periodate oxidation. After 18 hr, 50 to 70% of the aldehyde had disappeared from the lymphocyte membrane. Oxidized sialyl residues disappear completely after 60 hr of culture. This period corresponds to the de novo synthesis of sialic acid residues on the surface of periodate-activated cells. The two classes of oxidized sialyl-glycoconjugates were found to behave in different ways. In effect, our data showed that the aldehydes remaining at 18 hr are mainly located on the gangliosides, whereas the aldehyde moieties located on high m.w. glycoproteins disappear from the cell surface between 9 and 18 hr. This would suggest that the remaining aldehydes located on gangliosides are not directly involved in the expression of suppressive activity. PMID- 3027170 TI - Characterization of the C3 receptor induced by herpes simplex virus type 1 infection of human epidermal, endothelial, and A431 cells. AB - Herpes simplex virus type 1 (HSV-1) infection induces the appearance of viral analogues of human Fc IgG and C3 receptors on the surface of human cells. The virally induced C3 receptor(s) has been broadly defined as a C3b receptor, but its ligand binding characteristics have not been rigorously defined. In this study, human epidermal cells, A431 cells, and human umbilical vein endothelial cells infected with HSV-1 demonstrated rosetting with sheep erythrocytes (E) coated with IgG (E-IgG) or the complement components C3b (EAC3b) or iC3b (EAC3bi), but not with E-IgM, C4 (EAC14), C3d (EAC3d), or E alone. Rosetting was markedly enhanced by pretreatment of HSV-1-infected cells with neuraminidase. Unlike human C3 receptors, the HSV-1-induced C3 receptor was found to be trypsin resistant. To determine whether HSV-1 induced CR1-like receptors or CR3-like receptors, infected cells were pretreated with EDTA, which is known to inhibit native CR3 function. EDTA failed to prevent rosetting with EAC3bi. Furthermore, blocking studies using monoclonal antibodies against CR1 and CR3 revealed that the anti-CR1 antibody 5C11 consistently blocked EAC3b and EAC3bi rosetting with HSV-1-infected cells in a dose dependent manner, but monoclonal antibodies against CR3 did not. This study indicates that the HSV-1-induced C3 receptor is an analogue of CR1. PMID- 3027172 TI - Recognition of human polymorphonuclear leukocyte receptors for leukotriene B4 by rabbit anti-idiotypic antibodies to a mouse monoclonal antileukotriene B4. AB - Rabbit anti-idiotypic IgG antibodies to the combining site of a mouse monoclonal IgG2b antibody to leukotriene B4 (LTB4) cross-reacted with human polymorphonuclear (PMN) leukocyte receptors for LTB4. Anti-idiotypic IgG and Fab both inhibited the binding of [3H]LTB4, but not [3H]N-formylmethionyl leucylphenylalanine (fMLP), to PMN leukocytes with similar concentration-effect relationships, whereas neither nonimmune rabbit IgG nor Fab had any inhibitory activity. At a concentration of anti-idiotypic IgG that inhibited by 50% the binding of [3H] LTB4 to PMN leukocytes, the antibodies preferentially recognized high affinity receptors. Anti-idiotypic IgG and Fab inhibited PMN leukocyte chemotactic responses to LTB4, but not fMLP, with concentration-effect relationships resembling those characteristic of the inhibition of binding of [3H] LTB4, without altering the LTB4-induced release of beta-glucuronidase. Chemotaxis and increases in the cytoplasmic concentration of calcium equal in magnitude to those elicited by optimal concentrations of LTB4 were attained at respective concentrations of anti-idiotypic IgG equal to and 1/25 the level required for inhibition of binding of [3H]LTB4 by approximately 50%. Thus, the anti-idiotypic antibodies bound to PMN leukocyte receptors for LTB4 with a specificity, preference for high affinity sites, and capacity to alter PMN leukocyte functions that were similar to LTB4. PMID- 3027173 TI - Modulation of fibroblast proliferation by sulfidopeptide leukotrienes: effect of indomethacin. AB - Because infiltration of mononuclear cells and fibroblast proliferation are associated in chronic inflammatory lesions, we tested the hypothesis that leukotrienes (LT), a product of activated mononuclear cells, may modulate fibroblast growth. Proliferation of cultured human skin fibroblasts was estimated by [3H]thymidine incorporation and cell count at increasing concentrations (0.1 nM to 0.1 microM) of LTC4 or LTD4. LTC4 and LTD4 stimulated cell growth in a dose dependent manner only in the presence of 50 microM indomethacin. Under similar conditions, LTE4 but not LTB4 (0.1 microM) was active. Both asynchronous, growing cells and synchronous, quiescent cells were sensitive to LT when prostaglandin (PG) synthesis was suppressed by indomethacin. Other blockers of cyclooxygenase such as ibuprofen and aspirin exhibited identical permissive activity, and the effect of indomethacin was totally abolished by addition of PGE2. LTC4 modified neither [3H]arachidonic acid release from prelabeled fibroblasts nor PGE2 production by fibroblasts. These results demonstrate that the sulfidopeptide LT stimulate fibroblast proliferation only when the endogenous synthesis of PG is blocked, but they do not enhance the synthesis of PG in their target cells showing no evidence for a negative feed-back loop. Nevertheless, it seems likely that the initiation and development of the fibrotic process in the different tissues depends in part on the local balance between PG and LT productions. PMID- 3027175 TI - Analysis of immune function in herpes zoster patients: demonstration and characterization of suppressor cells. AB - We sought to identify imbalances of immune regulatory cells that might contribute to the depression of cell-mediated immunity that occurs during an episode of herpes zoster. Peripheral blood mononuclear cells (PBMC) were obtained from patients with herpes zoster during the acute (less than 7 days after disease onset) and convalescent (more than 10 days after disease onset) phases of illness and from healthy seropositive donors. The PBMC were analyzed for: lymphoproliferative responses to varicella-zoster virus (VZV) antigens, Leu-3 (helper/inducer):Leu-2 (cytotoxic/suppressor) ratios, and percentages of suppressor cells as defined by coexpression of the Leu-2 and OKM1 antigens. Significantly depressed proliferative responses of VZV antigens and Leu-3:Leu-2 ratios, and increased percentages of Leu-2+ OKM1+ suppressor cells were observed in PBMC of acute phase herpes zoster patients as compared with the PBMC of convalescent patients or healthy donors. These differences were also observed in individual patients sequentially studied during both phases of disease. Cryopreserved acute phase PBMC suppressed the proliferative response of autologous convalescent phase PBMC to VZV antigens, but not to herpes simplex virus (HSV) antigens. The acute phase PBMC suppressor cell was radiation sensitive and was identified as a Leu-2+ cell by fluorescence-activated cell sorting. Thus, depression of cell-mediated immunity during the acute phase of herpes zoster was associated with a relative increase of lymphocytes expressing a suppressor cell phenotype and the activation of a radiosensitive Leu-2+ suppressor cell with some degree of antigen specificity. PMID- 3027174 TI - Sensitivity of simian virus 40-transformed C57BL/6 mouse embryo fibroblasts to lysis by murine natural killer cells. AB - The susceptibility of mouse cells expressing full-length or truncated transforming protein (T antigen) of simian virus 40 (SV40) to lysis by murine natural killer (NK) cells was assessed. For these studies, C57BL/6 mouse embryo fibroblasts (B6/MEF) were transformed by transfection with SV40 DNA encoding the entire T antigen. The transformed cell lines were tested for susceptibility to lysis by nonimmune CBA splenocytes as a source of NK cells and to lysis by C57BL/6, SV40-specific cytolytic T cells (CTL). It was found that 13 of 15 clonally derived, SV40-transformed H-2b cell lines were susceptible to lysis by NK cells. However, there was some variation in their susceptibility to lysis by NK cells. There was no correlation between susceptibility to lysis by SV40 specific CTL and to lysis by NK cells. Cells transfected with a plasmid which encodes only the N-terminal half of the SV40 T antigen were consistently less susceptible to lysis by NK cells, suggesting that expression of only the N terminus of the T antigen was insufficient for optimal susceptibility to lysis by NK cells. Primary mouse embryo fibroblasts transformed by human adenovirus type 5 E1 region DNA were also found to be susceptible to NK cell-mediated lysis. Lysis of SV40-transformed cells by nonimmune CBA splenocytes was mediated by NK cells because: lysis was augmented when the effector cells were treated with interferon before assay; and lysis was abrogated when the effector cells were obtained from mice that had been depleted of NK activity by treatment with antiserum against the asialo GM1 surface marker. These results indicate that primary mouse cells which are transformed by SV40 and which express the native T antigen are susceptible to lysis by mouse NK cells. Conversely, cells transformed by a plasmid encoding only the N-terminal half of the T antigen express reduced susceptibility to lysis by NK cells. PMID- 3027176 TI - Reactivation of herpes simplex virus is associated with production of a low molecular weight factor that inhibits lymphokine activity in vitro. AB - Peripheral blood mononuclear cells obtained during recrudescent Herpes simplex virus (HSV) infection and stimulated with UV-inactivated viral antigen (UV-HSV) for 24 hr produced a low molecular weight (dialyzable) factor that inhibited lymphokine activity. This factor prevented the expression of leukocyte inhibitory factor (LIF) activity, but not its production. It was not made in UV-HSV stimulated cultures grown in presence of 2 X 10(-6) M indomethacin nor in cultures of peripheral blood mononuclear cells obtained during convalescence or quiescence (greater than 4 days from onset of clinical symptoms) or from seropositive controls without a history of recurrent HSV disease. Dialyzable inhibitory factor production required OKM1+, OKT8+, and OKIa+ cells as determined by complement-mediated lysis with monoclonal antibody. Dialyzable inhibitory factor activity was associated with a trypsin-sensitive 8.2 K fraction as determined by Sephadex chromatography followed by sodium dodecyl sulfate acrylamide gel electrophoresis. PMID- 3027177 TI - Identification of an interferon-gamma response region 5' of the human histocompatibility leukocyte antigen DR alpha chain gene which is active in human glioblastoma multiforme lines. AB - The expression of class II major histocompatibility (MHC) antigens is central to the mounting of a successful immune response. Understanding the molecular mechanisms involved in the induction of class II MHC expression may therefore provide the knowledge necessary to manipulate the immune system appropriately. We are particularly interested in the induction of class II MHC antigens on cells in the brain, because of the potential involvement of such brain cells in the initiation and perpetuation of autoimmune-like diseases of the central nervous system. We examined the mechanisms involved in interferon-gamma (IFN-gamma) induction of class II MHC antigens on a human glioblastoma multiforme cell line. This paper describes the identification of a 297-bp (base pair) fragment of the class II MHC DR alpha chain gene which is involved in IFN-gamma induction. We were able to identify this IFN-gamma responsive sequence by preparing recombinant plasmids containing 5' flanking pieces of the human DR alpha chain gene placed upstream of the indicator gene, chloramphenicol acetyltransferase (CAT). These recombinant plasmids were transfected into human glioma cells which were then cultured in the presence or absence of IFN-gamma. After 48 hr, transient expression of CAT was assayed by thin layer chromatography. CAT enzyme activity was significantly increased only in IFN-gamma-treated cells. This increase was also reflected at the RNA level in that IFN-gamma treatment resulted in higher CAT transcripts. A computer homology search revealed a possible consensus sequence shared among different IFN-gamma-inducible genes. PMID- 3027178 TI - In vitro regulation of IgA subclass production. III. Selective transformation of IgA1 producing cells by Epstein-Barr virus. AB - In past experiments, using limited dilution analysis, we have demonstrated that a high percentage of immunoglobulin-secreting clones derived from Epstein-Barr virus- (EBV) stimulated lymphocytes secrete IgA. To further characterize the IgA produced by these clones, the IgA subclass of supernatants from clones stimulated 4 to 6 wk previously with EBV was determined by radioimmunoassay. All of 17 IgA producing clones secreted IgA1; none secreted IgA2. Because we have shown that surface IgM+ (sIgM+) B cells are an enriched source of IgA2 plasma cell precursors, panning techniques were used to purify sIgM+ B cells from tonsils. Of 103 clones derived from these sIgM+ B cells, 102 secreted IgA1 and only one secreted IgA2. The relative absence of IgA2-producing clones could not be attributed to an absence of EBV receptors on IgA2 cells. A mean of 84 +/- 4% of freshly isolated IgA2 B cells and 78 +/- 6% of IgA1 B cells could be stained with a monoclonal antibody binding the EBV receptor; and there was no failure of EBV to infect IgA2 plasma cells precursors. Of IgA2 plasma cells derived from peripheral blood lymphocytes stimulated 7 days previously with EBV, 54 +/- 7% were positive for the EBV nuclear antigen, compared with 54 +/- 18% of IgA1 plasma cells from the same cultures. Seven days after EBV stimulation, a mean of 25% of the total IgA plasma cells were positive for cytoplasmic IgA2, whereas by 21 days after stimulation only 7% were positive for IgA2. This shift in the proportions of IgA1 and IgA2 plasma cells could be attributed to a failure of the IgA2 plasma cell number to increase after 10 days in culture. There was no evidence for selective suppression of IgA2 production by T cells or selective lysis of IgA2 plasma cells by infectious EBV particles. These results demonstrate that although precursors for both IgA1- and IgA2-producing cells can be stimulated to differentiate in response to EBV, there is preferential transformation of IgA1-producing cells. PMID- 3027179 TI - Variations in genetic control of susceptibility to Theiler's murine encephalomyelitis virus (TMEV)-induced demyelinating disease. I. Differences between susceptible SJL/J and resistant BALB/c strains map near the T cell beta chain constant gene on chromosome 6. AB - In some susceptible mouse strains, intracerebral (IC) inoculation of Theiler's murine encephalomyelitis virus (TMEV) results in a persistent infection leading to chronic demyelinating disease. Previous genetic analyses between susceptible SJL/J and resistant C57BL/6 mice indicated a role for multiple unlinked genes in the development of clinical and histopathological disease, including a major influence of the D region of the H-2 complex. In this study, genetic analysis of a different strain combination (susceptible SJL/J and resistant BALB/c) also demonstrates the involvement of multiple genes, but the H-2 genotype (H-2s and H 2d, respectively) does not appear to contribute significantly to susceptibility differences. In both segregation studies and recombinant-inbred (R-I) analysis, clinical and histopathological disease occurs in both H-2s homozygotes and H-2d homozygotes (as well as H-2s/H-2d heterozygotes), with the actual frequency related to the proportion of non-H-2 genome from the susceptible strain. There appear to be at least two non-H-2 genes involved in differential susceptibility of SJL/J and BALB/c to TMEV-induced disease. Analysis of R-I strains generated from BALB/c and SJL/J progenitors indicates linkage of at least one of these non H-2 genes to those encoding the constant portion of the beta-chain of the T cell receptor on chromosome 6. Many genes may actually be involved, but each strain comparison defines a different subset of these loci--only those at which the two strains in question carry "functionally" different alleles. Thus, different strain comparisons may accent the roles of different genes in resistance to the same infectious organism or disease process. In addition to the genes identified thus far, there may be yet other genes contributing to development of TMEV induced disease, but their recognition may require analysis of still other strain combinations. PMID- 3027180 TI - Effects of leukocyte inhibitory factor (LIF) on neutrophil mediated antibody dependent cellular cytotoxicity. AB - The human lymphokine, leukocyte inhibitory factor (LIF), was investigated for its effect on neutrophil-mediated antibody-dependent cellular cytotoxicity (ADCC) for K562 targets. Highly purified LIF (0.5 to 2 U/ml) induced a significant dose dependent potentiation of neutrophil ADCC by up to 54.9% (p less than 0.001). Higher concentrations of LIF inhibited cytotoxicity. The degree of cytotoxicity was found to correlate (r = 0.99) with the increased secretion of superoxide after neutrophil-target cell interaction. Anaerobic conditions inhibited cytotoxicity mediated by both control and LIF-treated neutrophils. The latter observation lends support to the concept that enhanced ADCC was mediated through increased superoxide production and not through the induction of a separate pathway. Increased superoxide production may have resulted from an upregulation of the transduction mechanism leading to neutrophil stimulation through the Fc receptor. In addition, we demonstrated an increased capacity of the neutrophil to adhere to its target (average 3.3:1 effector:target ratio in untreated cells to 4.8:1 after treatment with LIF), and this may also have been responsible for the increase in the respiratory burst and subsequent enhanced ADCC. These observations provide potential support for an in vivo role for LIF in tumor immunity. PMID- 3027181 TI - Enhanced 5-lipoxygenase activity in lung macrophages compared to monocytes from normal subjects. AB - We compared lipoxygenase activities of lung macrophages obtained from bronchoalveolar lavage to activities of blood monocytes purified by using discontinuous plasma/Percoll density gradients and adherence to tissue culture plastic in five normal subjects. Cells were incubated with ionophore A23187 (10( 9) to 10(-5) M) or arachidonic acid (0.12 to 80 microM) for 1 to 60 min at 37 degrees C to construct dose-response and time-dependence curves of lipoxygenase product generation. Products were identified and were quantified by using high pressure liquid chromatography and ultraviolet spectroscopy. Under all conditions of product generation, both macrophages and monocytes generated predominantly (5S,12R)-dihydroxy-(6Z, 8E, 10E, 14Z)-eicosatetraenoic acid (leukotriene B4 (LTB4] and (5S)-hydroxy-(6E, 8Z, 11Z, 14Z) - eicosatetraenoic acid (5 - HETE), but, in each subject, macrophages invariably released greater amounts of LTB4 and 5-HETE than monocytes. In response to A23187, macrophages released a maximum of 183 +/- 96 pmol of LTB4 and 168 +/- 108 pmol of 5-HETE per 10(6) cells (mean +/- SEM), whereas monocytes released only 16 +/- 1 and 18 +/- 8 pmol per 10(6) cells of LTB4 and 5-HETE, respectively. After adding arachidonic acid, macrophages released a maximum of 52 +/- 21 pmol of LTB4 and 223 +/- 66 pmol of 5-HETE, whereas monocytes released no detectable products. The results suggest that mononuclear phagocyte maturation in the lung may be accompanied by an enhanced ability to generate 5-lipoxygenase products. PMID- 3027182 TI - Induction of anti-AKR/gross virus cytolytic T lymphocytes in AKR.H-2b:Fv-1b congenic mice: age-dependent conversion to a nonresponder phenotype. AB - Previously, we reported that the generation of cytolytic T lymphocytes (CTL) specific for syngeneic tumors induced by AKR/Gross leukemia viruses was under multi-gene control. Thus, although carrying the required immune response gene(s) encoded by the H-2b haplotype and characteristic of responder strains such as C57BL/6, AKR.H-2b congenic mice failed to mount antiviral CTL responses. Young adult AKR.H-2b:Fv-1b "doubly congenic" mice, however, were able to generate specific anti-AKR/Gross virus CTL activity. These results demonstrated that the positive effect of MHC-encoded immune response gene control could be overcome by the action of the Fv-1n allele. The responder status of the B6.Fv-1n congenic, however, indicated that this Fv-1n-mediated inhibition was dependent on the interaction of Fv-1n with another gene(s) encoded by the AKR background. The results of experiments performed with AKXL recombinant inbred mice further suggested that a single additional genetic locus, encoding the Akv-1 provirus, was necessary along with Fv-1n to cause inhibition of antiviral CTL generation. Here we show that the responsiveness of AKR.H-2b:Fv-1b mice is dependent on their age. Thus, with moderate aging these doubly congenic mice converted to a nonresponder status with respect to anti-AKR/Gross virus CTL production: 85% of mice less than or equal to 9 wk of age responded compared with 0% of mice greater than 9 wk old. As with nonresponder AKR.H-2b mice, an inverse correlation was observed between CTL responsiveness and the expression of CTL-defined viral antigens by normal cells. Namely, spleen cells from young AKR.H-2b:Fv-1b mice showed little or no expression of such viral antigens, whereas with moderate aging there was a steady increase in their display. These results are discussed with reference to possible mechanisms of unresponsiveness of AKR.H-2b vs moderately aged AKR.H-2b:Fv-1b mice, and with respect to the utility of this system as a model for naturally occurring retrovirus infections and the interactions of retroviruses with the immune system. PMID- 3027183 TI - Re.: Surface antigen preservation by PLP and formalin fixation and the effects of tissue processing. PMID- 3027184 TI - Specific purification of monoclonal anti-DNA antibodies from culture medium using a DNA-coupled Sepharose 4B affinity column. AB - Human monoclonal antibodies against DNA were specifically purified from the culture medium of an EBV transformant of SLE patients' lymphocytes using a DNA coupled Sepharose 4B affinity column. The monoclonal antibodies were eluted from the column with 5% dimethylsulfoxide (pH 10.7) containing 0.5 M NaCl without loss of immunological activity and without contamination by other proteins. PMID- 3027185 TI - Detection of influenza viruses through selective adsorption and detection of the M-protein antigen. AB - A model system has been developed which permits rapid detection of influenza viruses through targeting of the M (membrane or matrix)-protein; a type-specific antigen, in an enzyme-linked immunosorbent assay system. This technique exploits the hydrophobic properties of M-protein; the M-protein is selectively and rapidly adsorbed to polystyrene surfaces even in the presence of a 5000-fold excess of bovine serum albumin. Hyperimmune antiserum prepared to purified M-protein is used as the detecting reagent. All type A influenza viruses could be detected by this technique, type B influenza viruses reacted to a slight extent and Sendai virus (parainfluenza virus, type 1) did not react. Virus could be detected to levels as low as 3 ng. Purification of M-protein and preparation of hyperimmune sera from other related virus groups, such as type B influenza viruses, paramyxoviruses and rhabdoviruses should permit detection of these agents by a similar technique. PMID- 3027186 TI - Antiviral activity of Bordetella pertussis vaccine-elicited peritoneal exudate cells. AB - Peritoneal exudate cells collected from mice 7 days after treatment with Bordetella pertussis vaccine exhibited significant in vitro antiviral activity against vesicular stomatitis virus (VSV). Vaccine-induced peritoneal exudate cells exhibited both intrinsic and extrinsic antiviral activity in culture with target VSV-infected L cells. Virus replication was poor in the vaccine-induced exudate cells. Coculture of vaccine-induced exudate cells and VSV-infected L cell targets decreased virus yield. The activity appeared specific for infected cells and at least a portion of the antiviral activity was directed against the initial infection cycle. Nonadherent vaccine-induced exudate cells showed an increase in antiviral activity over total vaccine-induced exudate cells. PMID- 3027187 TI - Morphological and functional changes in mouse splenic lymphocytes following in vivo and in vitro exposure to chlorphentermine. AB - With repeated administration to animals, the cationic, amphiphilic drug, chlorphentermine (CP), has been shown by others to induce a phospholipidosis in lymphocytes. In the present study mouse splenic lymphocytes, exposed to CP, either in vivo or in vitro, developed morphological changes consistant with the induction of phospholipidosis. In addition, CP induced functional changes in lymphocytes. Mice, treated with CP in vivo, demonstrated a significantly depressed ability to generate a delayed hypersensitivity response or to produce antibody-secreting cells against de novo antigens. Mouse splenic lymphocytes, exposed to 10(-7) M CP for 3 days in vitro, demonstrated a significantly depressed blastogenic response to the mitogens phytohemagglutinin, concanavalin A and lipopolysaccharide. CP inhibited an event that occurred early during lymphocyte activation, but was subsequent to mitogen/receptor coupling. In addition, CP significantly depressed the increased uptake of choline that occurs in lymphocytes following cellular activation. Since the presence of phospholipidosis is indicative of an impairment in phospholipid metabolism, these results taken together provide evidence for a relationship between this phenomenon and altered immune function. PMID- 3027188 TI - Effects of chronic intracutaneous administration of arachidonic acid and its metabolites. Induction of leukocytoclastic vasculitis by leukotriene B4 and 12 hydroxyeicosatetraenoic acid and its prevention by prostaglandin E2. AB - Despite the postulated role of arachidonic acid-derived metabolites in the pathophysiology of chronic inflammatory dermatoses such as psoriasis and atopic or contact dermatitis, the cutaneous effects of their chronic application have not yet been investigated. We therefore studied systematically the effects of chronic intracutaneous administration of arachidonic acid, prostaglandin E2 (PGE2), leukotriene B4 (LTB4), and 12-hydroxyeicosatetraenoic acid (12-HETE) in guinea pigs, and describe previously unrecognized findings partly different from those reported in the past for short-term or topical application of these inflammatory mediators. Leukotriene B4 and 12-HETE led to massive histologic changes characteristic of leukocytoclastic vasculitis. These changes could be prevented by concomitant PGE2 administration. In epidermis, LTB4 and 12-HETE caused some spongiosis as well as hyperplasia and increased tritiated thymidine autoradiographic labeling index. Arachidonic acid and PGE2 alone had little effect. These data suggest that in addition to other inflammatory or hyperproliferative dermatoses, arachidonic acid metabolites formed via lipoxygenase pathways could play a major role in leukocytoclastic vasculitis, whereas PGs could exert a tissue-protective effect. PMID- 3027189 TI - A new type of human papillomavirus associated with oral focal epithelial hyperplasia. AB - Lesions from 10 patients suffering from focal epithelial hyperplasia (FEH) of the oral mucosa, including those of 4 Greenlandic Eskimos, were investigated for the presence of human papillomavirus (HPV) DNA sequences by blot hybridization experiments. Two distinct HPVs were detected in the DNA extracted from these lesions, and their genomes were molecularly cloned and characterized. One of these HPVs, detected in 4 patients, was found to be identical with HPV13, whose association with FEH was already known. The other one, detected in 6 patients, was only weakly related to HPV13 and to the other HPVs associated with lesions of the mucous membranes, and constituted a new HPV type, tentatively named HPV32. Lesions from other types of oral papillomas, obtained from 14 additional patients, were also analyzed. Human papillomavirus DNA sequences were detected in the DNA preparations extracted from 5 specimens: HPV6 DNA in a condyloma and in a papilloma, 2 as yet uncharacterized HPV DNAs in 2 papillomas, and HPV32 DNA in a papilloma which showed histologic similarities to FEH. Thus, it seems likely that FEH of the oral mucosa is a disease associated with 2 specific HPVs--HPV13 and HPV32. PMID- 3027191 TI - Transmission of infection with herpes simplex virus by renal transplantation. AB - Disseminated infection with herpes simplex virus type 2 was identified in two patients 20 days after they had received kidney transplants from the same organ donor. Neither patient had neutralizing antibody to herpes simplex virus before transplantation, and both had herpes simplex virus isolated from surveillance cultures of urine before the onset of clinical symptoms. A clear focus of primary infection was not found in either patient. Analysis of the patients' isolates by DNA restriction endonuclease analysis strongly suggested that the strains were identical. These data implicate the allografts as the source of the viral infection. PMID- 3027190 TI - PIXE analysis in different stages of psoriatic skin. AB - Elemental distribution in psoriatic skin varies with the functional state of the keratinocytes, e.g., electrolytes influence cell metabolism and cell proliferation, and trace elements play a crucial role in a great number of enzymes. Elemental distribution in pinpoint lesions, old plaques, and uninvolved skin of 5 psoriatic patients and 4 healthy controls was studied by means of PIXE (proton-induced x-ray emission) analysis. This technique allows the simultaneous detection of elements with an atomic number greater than or equal to 14 along the epidermis and dermis in freeze-dried skin biopsies. Trace elements such as Fe, Cu, and Zn were determined down to a level of 1 ppm. In comparison with uninvolved skin, concentrations of P and K were elevated in psoriatic epidermis. In addition, increased levels of K were correlated with the stage of the psoriatic lesion. Zinc concentrations were significantly elevated in pinpoint lesions. The Zn concentration profiles within the epidermis and upper dermis showed high correlation to the P concentration profiles. Iron levels were decreased in old psoriatic plaques, whereas Cu concentrations varied considerably. In comparison to the controls, Cl concentrations were markedly decreased in the dermis of involved and uninvolved psoriatic skin, whereas epidermal Cl levels were unaffected. As high K levels prevent the Ca-induced differentiation of keratinocytes, high K levels may be the cause of the high cell differentiation in psoriatic skin. Elevated DNA- and RNA-polymerases might be the cause of elevated Zn levels in pinpoint lesions. PMID- 3027192 TI - Murine cytomegalovirus genomic material in marrow cells: relation to altered leukocyte counts during sublethal infection of mice. AB - To investigate infection of hematopoietic cells by murine cytomegalovirus (MCMV), we used in situ hybridization to detect MCMV genomic material in marrow cells during sublethal infection of three-week-old mice. MCMV genomic material was present in femoral marrow cells on days 3, 5, 7, and 14, and detection of MCMV genomes correlated strongly with recovery of infectious virus. Peak infection occurred on day 5, when 0.003%-0.185% of the marrow cells contained MCMV genomic material. At peak infection, MCMV genomic material was observed predominantly in marrow mononuclear cells. Coincident with marrow infection, mice experienced significant leukopenia, whereas cessation of MCMV replication in marrow cells was associated with leukocytosis and restoration of normal peripheral blood leukocyte counts. PMID- 3027193 TI - DNA-DNA dot hybridization to detect Epstein-Barr virus in throat washings. AB - To compare the abilities of the nucleic acid dot hybridization assay and the cord blood lymphocyte transformation assay to detect Epstein-Barr virus (EBV), we examined throat washings from healthy control subjects (nine EBV-seronegative and 51 EBV-seropositive), patients with acute infectious mononucleosis, and renal transplant recipients. The dot hybridization assay detected EBV excretion in four (8%) of the EBV-seropositive controls; three of these four were also positive by the lymphocyte transformation assay. Throat washings from seven (87.5%) of eight patients with acute infectious mononucleosis were positive by both assays. EBV was present in throat washings from 13 (50%) of 26 renal transplant recipients. For specimens stored at -70 C for less than four months, the dot hybridization assay had a sensitivity of 90% and a specificity of 98% when compared with the lymphocyte transformation assay. The dot hybridization assay is a rapid, sensitive, and specific test that can be performed on readily available clinical specimens. PMID- 3027194 TI - Restriction endonuclease patterns of cytomegalovirus after passage in humans. PMID- 3027195 TI - Use of ganciclovir to treat serious cytomegalovirus infections in patients with AIDS. PMID- 3027196 TI - Reactogenicity and antigenicity of the rhesus rotavirus vaccine in Venezuelan children. PMID- 3027197 TI - Rhesus rotavirus vaccine in infants and children. PMID- 3027198 TI - The pathogenesis of acute viral encephalitis and postinfectious encephalomyelitis. AB - Japanese encephalitis and postmeasles encephalomyelitis represent two important forms of acute encephalitis in the developing world. Japanese encephalitis accounts for 20,000 acute illnesses per year, and measles encephalomyelitis accounts for as many as 100,000. Both are accompanied by mortalities of 20%-30%, and in both forms, approximately one-half of the survivors have neurological sequelae. Therefore, these two diseases are responsible for a considerable proportion of worldwide mental and motor disabilities. Furthermore, both diseases are preventable with use of safe vaccines. Despite these similarities, their very different pathologies appear to be based on different mechanisms of pathogenesis (table 2). In Japanese encephalitis there is direct invasion of the virus into the nervous system, selective infection and destruction of neurons, and evidence that both humoral and cellular immune responses attenuate the infection. In measles encephalomyelitis there is little evidence that the virus invades the nervous system. Rather, measles virus infects lymphoid cells and causes abnormalities in immune regulation. Humoral immune responses are not found within the nervous system, and the cellular immune response may be inappropriately directed against host antigens. In measles encephalomyelitis, the inflammatory cells entering the nervous system may be the effector cells of autoimmune disease. PMID- 3027200 TI - Evaluation of three assays on alveolar lavage fluid in the diagnosis of cytomegalovirus pneumonitis after bone marrow transplantation. AB - Cytologic examination, immunofluorescence assays, and cultures for virus were compared prospectively in the detection of cytomegalovirus (CMV) in cells obtained by 41 bronchoalveolar lavages (BALs) from 30 bone marrow transplant recipients with pneumonia. No evidence of CMV was found in 21 BALs. Viral inclusions, positive immunologic assays, and positive cultures were found in 14, 15, and 18 BALs, respectively. Cytological and immunologic results were closely related except in one BAL. In five BALs, a positive culture for virus was the sole criterion noted for diagnosis of CMV. The patients with both cytologically and immunologically positive results were more likely to die with or from respiratory failure (P less than .05), diffuse interstitial patterns (P less than .01), and severe acute graft-vs.-host disease (P less than .02) than were the patients without any criterion for diagnosis of CMV. The discrepancies with the previously reported data, the interpretation of the three diagnostic procedures, and the ability of BAL to diagnose CMV pneumonia are discussed. PMID- 3027202 TI - Transmission of cytomegalovirus by pretransplant leukocyte transfusions in renal transplant candidates. PMID- 3027199 TI - Novel agents of viral enteritis in humans. AB - Considerable information has recently emerged regarding certain "groups" of novel viruses associated with gastroenteritis in humans. The viruses reviewed here are 20-35 nm in diameter and can be detected in the stools of acutely ill individuals with gastroenteritis. These viruses can be conveniently divided into four "groups": Norwalk-like agents, caliciviruses, astroviruses, and other small round viruses. The evidence for the etiologic association of these agents with gastroenteritis varies from agent to agent but is most extensive for the Norwalk like agents, of which the Norwalk serotype is a major cause of epidemic gastroenteritis. Human caliciviruses appear to be relatively common causes of gastroenteritis in children, particularly in Japan and the United Kingdom. Astroviruses have been reported as occasional causes of gastroenteritis in children and adults in various parts of the world. The epidemiological significance of the other small round viruses is unknown. Because of the difficulty of cultivating these agents in vitro, biochemical and antigenic characterization of these agents is incomplete. An understanding of the pathogenesis of disease and of the roles of immune responses to infection is similarly at a primitive stage. The recent development of sensitive and efficient assays for detection of several of these agents and the reports that certain strains of human caliciviruses and astroviruses can be cultivated in vitro should facilitate characterization and epidemiological studies of these agents. Systematic, prospective epidemiological studies of these agents in well-defined populations of various age groups are sorely needed for definition of the relative importance of each agent in human disease. Such information is essential for the consideration of appropriate control measures. PMID- 3027203 TI - Transgenic mice and the language of cells. PMID- 3027201 TI - Quantitation of human cytomegalovirus DNA in lungs from bone marrow transplant recipients with interstitial pneumonia. AB - DNA-DNA hybridization was compared with culture for virus and histopathology for the detection and quantitation of human cytomegalovirus (HCMV) in lungs from bone marrow transplant recipients. The hybridization assay detected 100 pg of HCMV DNA or greater than 520 plaque-forming unit equivalents. Compared with culture for virus, the hybridization assay had a sensitivity of 90.6% and a specificity of 77.8%. The quantity of HCMV DNA and the infectivity titer were directly correlated. Quantitation of HCMV DNA by hybridization was more reproducible than were infectivity titers for frozen specimens. Histopathologic examination of lung specimens detected HCMV inclusions in 60% (15 of 25) of patients with HCMV associated interstitial pneumonia, whereas hybridization results were positive for 96% (24 of 25) of these patients. A significant inverse correlation was demonstrated between quantitative values for HCMV DNA and the duration of pneumonia. PMID- 3027204 TI - Concomitant enhancement of a cytoskeleton-associated 76,000-dalton protein and inhibition of fluid-phase pinocytosis by interferon-alpha in Fujinami sarcoma virus-transformed rat 3Y1 cells. AB - Addition of rat interferon-alpha (IFN-alpha) to Fujinami sarcoma virus transformed rat 3Y1 cells progressively inhibited fluid-phase pinocytosis [10% inhibition at 3 h; maximal (60%) inhibition by 12 h]. Electrophoretic analysis of the cytoskeletal fraction from cultures exposed to IFN for 24 h revealed a novel 76,000-dalton protein (CKp76). The kinetics of its appearance paralleled the inhibition of fluid-phase pinocytosis. CKp76 was not detected in cultures pretreated with actinomycin D, or prelabeled with [35S]methionine, prior to IFN addition. However, the presence of cycloheximide during incubation with IFN had no effect on the synthesis of CKp76 after removal of both agents. These results suggest that the appearance of CKp76 was due to enhanced transcription of its gene in response to IFN. Subcellular fractionation revealed the presence of induced CKp76 in the nuclear pellet. From these results it is possible that CKp76 may be responsible at leat in part for the effects of IFN on fluid-phase pinocytosis. PMID- 3027205 TI - A study of calcification in the leg tendons from the domestic turkey. AB - Details are presented of the structure and the spatial and temporal sequence of calcification within leg tendons obtained from the domestic turkey, Meleagris gallopavo. Histological and ultrastructural examination of the tissues reveals the development of large tendonoblasts, disposed in columns parallel to tendon long axes. Maturing cells are characterized by organelles active in protein synthesis, few mitochondrial granules, and an extensive system of processes. Deposition of mineral in the extracellular tissue spaces appears initially associated with small vesicular structures located between collagen fibrils. As calcification progresses, mineral is associated predominantly with collagen. Electron diffraction and electron probe microanalysis indicate that the mineral is a poorly crystalline hydroxyapatite having a Ca/P molar ratio ranging between 0.9 and 1.5. Following decalcification and staining of tissue sections, vesicular structures retain integrity by uptake of lead and uranium salts while collagen stains only poorly. Implications of these results are discussed with respect to the physicochemical and biological events of vertebrate calcification. PMID- 3027206 TI - Evaluation of nerve function deficit. Its improvement by nerve decompression or corticosteroid therapy. AB - An attempt has been made to use the protocol for evaluation of nerve damage prepared at a "Workshop on Neuritis" held at Schiefflein Leprosy Research and Training Centre, Karigiri in three cases of ulnar nerve decompression, three cases of median nerve decompression and three cases of ulnar neuritis on corticosteroid therapy. The nerve function deficit did show an improvement. The problems and suggestions are presented with a view to encourage the use of the said protocol to enable comparison of data amongst different centres and different therapy. PMID- 3027207 TI - Use of preformed hydroxylapatite blocks for grafting in genioplasty procedures. AB - Dense hydroxylapatite is a synthetic, biocompatible, immunologically inert material that can establish a chemical union with bone when placed in intimate contact in histologic studies. The following is a preliminary report of the use of preformed dense, non-resorbable hydroxylapatite blocks as a grafting material for use in advancement and vertical lengthening of the bony chin associated with orthognathic surgical procedures. In rare instances where an autogenous bone graft would be required for vertical lengthening without any advancement of bony pogonion, the need is obviated by utilization of the hydroxylapatite solid block. PMID- 3027208 TI - Collagen tubes: role in subperiosteal contour augmentation. AB - Collagen-contained implants of particulate hydroxylapatite (C-HA) were compared to implants of hydroxylapatite alone (HA). The collagen-contained implant was more rapidly and more firmly attached to the recipient site and demonstrated more consistent particle consolidation than implants of HA alone. Additionally, the collagen did not interfere with the favorable properties of the particulate hydroxylapatite. Results are excellent for the (C-HA) implant and warrant further investigation in both animals and in man. PMID- 3027210 TI - [Antiviral effect of glycyrrhizin on varicella-zoster virus replication in vitro]. PMID- 3027211 TI - [Improved surgical management in Poland syndrome]. PMID- 3027209 TI - Pleomorphic adenoma of the cheek. AB - The mucosa of the cheek is a relatively rare site of occurrence for intraoral pleomorphic adenoma. A case of a large pleomorphic adenoma of the cheek is reported and a review of this lesion in the cheek is presented. PMID- 3027213 TI - [Reorganization of human ovarian adenocarcinoma cells and endometrial adenocarcinoma cells cultured in double-layered floating collagen gels]. AB - Collagen gels have been used as the substratum for epithelial cell culture to maintain their differentiated states. In this report, we proposed a double layered floating collagen gel method for the culture of human carcinoma cells to show their differentiated structures which closely resembled those in the tumors in vivo. On the 3rd week of the culture, OMC-1 cells (human ovarian mucinous adenocarcinoma) showed organized ductular structures. PAS-positive substances were seen along the surface of the ductules and also in cytoplasms. Electron microscopically, numerous microvilli were observed at the apical surfaces of the cells, with formation of rootlets. Ishikawa line cells (human well-differentiated endometrial adenocarcinoma) formed tubular structures with back-to-back arrangements. Electron microscopically, numerous microvilli, well-developed junctional complexes, and well-developed mitochondria and Golgi apparatus were observed, similarly to those in the cells grown in vivo. SNG-M cells (human moderately-differentiated endometrial adenocarcinoma) were arranged in sheets with occasional formation of tubular structures. Mitochondria, Golgi apparatus and desmosomes were poorly developed in general; however, the cells forming tubular structures showed signs of cellular differentiation. The floating and non floating methods were compared with the result that the former method was seen to be far superior to the latter for expressing cellular differentiation. PMID- 3027212 TI - [Endocrinological and clinico-pathologic study of granulosa-theca cell tumors of the ovary]. AB - Nine granulosa-theca cell tumors (four pure theca cell tumors, one granulosa cell tumors, two granulosa-theca tumors and two juvenile granulosa-theca tumors) were studied endocrinologically and clinico-pathologically. The cases of juvenile granulosa-theca tumors developed precocious pseudopuberty. Three of seven other cases were re-feminized and four cases showed no hormonal manifestation clinically. The peripheral vein serum values of estradiol, progesterone, testosterone and prolactin were elevated in six of eight cases, three of four cases, four of six cases, and two of three cases, respectively. The concentration ratios between tumor harboring ovarian vein samples and peripheral vein (or opposite normal ovarian vein) samples was 2.7 to 16.9 for estrone, 8.8 to 28.6 for estradiol, 3.6 to 4.7 for progesterone, 1.6 to 6.6 for testosterone and 0.6 to 1.0 for prolactin. Estradiol was localized in both granulosa cells and theca cells, and testosterone was localized in granulosa cells in half of the cases and in theca cells in 60% of the cases. Also, testosterone was localized in all three cases in which luteinized theca cells were present. There were no cases with positive prolactin localization. These results are compatible with the concept that in granulosa-theca cell tumor, both granulosa and theca cells can produce a wide range of steroid hormones. PMID- 3027214 TI - [Recent medical topics on platelets in hemorrhagic and thrombotic diseases]. PMID- 3027215 TI - Antithrombotic activity of glomerular adenosine diphosphatase in the glomerular basement membrane of the rat kidney. AB - Activity of nucleoside polyphosphatases (including adenosine diphosphatase [ADPase]) in the glomerular basement membrane (GBM) of the rat kidney can be demonstrated in situ by using cytochemical methods at the ultrastructural level. To study the possible influence of glomerular ADPase activity on experimentally induced intraglomerular platelet aggregation, we carried out alternate perfusion experiments with human platelets and adenosine diphosphate (ADP) solution in rat kidneys ex vivo. This was done in rats with reduced glomerular phosphatase activity induced by either an intravenous injection of doxorubicin (8.5 mg/kg body weight) or local x-irradiation (2000 rads) as well as in rats with normal glomerular enzyme activity, that is, untreated rats or rats injected intravenously with aminonucleoside of puromycin (PAN) (15 mg/kg body weight). It is shown that in kidneys of both doxorubicin-treated and x-irradiated rats intraglomerular platelet aggregation occurs in approximately 50% of the glomeruli, whereas in PAN-treated or control rats no platelet aggregation could be detected by light microscopy. Activated platelets (by electron microscopy) and beta-thromboglobulin or platelet factor 4 (immunofluorescence microscopy) could be detected with appropriate fluorescinated antibodies along the GBM exclusively in kidneys with reduced ADPase activity caused by doxorubicin or x-irradiation treatment. Because glomerular ADPase activity in contrast to other putative antithrombotic molecules in the GBM, that is, heparan sulfate proteoglycans, is clearly affected by doxorubicin or x-irradiation treatment, it is suggested that the activity of glomerular ADPase may reflect an important antithrombotic principle in the GBM of the rat kidney. PMID- 3027216 TI - Respiratory failure secondary to Eaton-Lambert syndrome. PMID- 3027217 TI - Intracellular pH changes during neutrophil activation: Na+/H+ antiport. AB - Activation of the Na+/H+ antiport mechanism was studied in human neutrophils by monitoring intracellular pH with a carboxyfluorescein derivative. N-formyl methionyl-leucyl-phenylalanine (FMLP) and phospholipase C (PLC) induced biphasic pH changes. Amiloride, which inhibits the antiport, completely blocked alkalinization but enhanced acidification. Polymyxin B, which inhibits protein kinase C, only blocked alkalinization. Activation with phorbol 12-myristate 13 acetate (PMA) led to alkalinization only; this was inhibited by amiloride or polymyxin B. Thus, during polymorphonuclear leukocyte (PMN) activation, intracellular alkalinization appears to be mediated by an amiloride-sensitive Na+/H+ antiport. Antiport activity can also be blocked indirectly by inhibition of protein kinase C activity. Early intracellular acidification does not appear to require kinase activity but is observed when phospholipids are remodeled with PLC. The antiport was also activatable by hypertonic buffered media. This response did not appear to be mediated by protein kinase C because it was unaffected by polymyxin B. Finally, superoxide generation was investigated. It is affected by, but not soley controlled by, either antiport or protein kinase C activity. PMID- 3027218 TI - Relationship of glucocorticoid suppression of arachidonic acid metabolism to alteration of neutrophil function. AB - The role of arachidonic acid metabolism in glucocorticoid-induced suppression of neutrophil function was investigated. In vivo treatment of cattle with dexamethasone decreased the production of lipoxygenase products of arachidonic acid metabolism (leukotriene B4 [LTB4] and H-5-hydroxy-6,8,11,14-eicosatetraenoic acid [5-HETE]) in neutrophils. We previously reported that in vivo dexamethasone treatment resulted in alteration of in vitro neutrophil oxidative metabolism, iodination, antibody-dependent cell-mediated cytotoxicity (ADCC), and random migration. To determine if the decrease in production of LTB4 and 5-HETE were responsible for the changes in neutrophil function, neutrophils were treated in vitro with inhibitors of the lipoxygenase enzyme (BW755c and nordihydroguaiaretic acid), and their function was evaluated. A decrease in neutrophil oxidative metabolism and iodination was observed, but there was no effect on neutrophil random migration or ADCC. The results suggest that in vivo treatment with glucocorticoids inhibit neutrophil oxidative metabolism and iodination by decreasing the formation of lipoxygenase products of arachidonic acid metabolism but alter neutrophil random migration and ADCC by a mechanism independent of arachidonic acid metabolism. PMID- 3027219 TI - 1,25-dihydroxyvitamin D3 stimulates interleukin-2 production by a T cell lymphoma line (MLA-144) cultured in vitamin D-deficient rat serum. AB - A lymphocyte T cell line (MLA-144), which constitutively secretes interleukin-2 (IL-2), was shown to express receptors for 1,25-dihydroxyvitamin D3 (1,25(OH)2D3). The proliferation of an IL-2-dependent cell line (HT-2) in response to supernatants from MLA-144 cells was employed as an index of IL-2 production by MLA-144 cells. IL-2 production was two fold higher from MLA-144 cells cultured in 2% vitamin D-deficient rat serum compared to 10% fetal calf serum (FCS). The addition of 1,25(OH)2D3 at 10(-15) M or 10(-11) M augmented IL-2 production by MLA-144 cells in vitamin D-deficient rat serum, but not in fetal calf serum. At 10(-7) M 1,25(OH)2D3 there was inhibition of IL-2 production by MLA-144 cells in either vitamin D-deficient serum or FCS. There was no effect of 1,25(OH)2D3 added directly to HT-2 cells. Monoclonal antibody to the IL-2 receptor competitively inhibited the proliferation of HT-2 cells in response to MLA-144 supernatants, suggesting that it was IL-2 from the MLA-144 supernatants which influenced HT-2 proliferation. Our findings demonstrate biphasic dose effects of 1,25(OH)2D3 on lymphokine secretion. The use of vitamin D-deficient rat serum allowed us to demonstrate the effects of 1,25(OH)2D3 in the physiologic and subphysiologic range. PMID- 3027220 TI - Pathophysiological mechanisms of altered transmembrane potentials in diseased human atria. AB - The pathophysiological mechanisms of altered transmembrane potentials in diseased human atria were investigated in 20 patients who were divided into group A (normal) and group B (diseased). The electrophysiological data of right atrial tissues measured with glass microelectrodes included maximum diastolic potentials (MDP), action potential amplitudes (APA) and action potential durations at the time required for 50% repolarization (ADP 50%) and 75% repolarization (APD 75%). The sarcolemma isolated from atrial tissues was used for determination of the sarcolemmal Na+-K+ ATPase activities. Anionic molecular sites distributed in the sarcolemmal complex were characterized by cationized ferritins (CF). The electrophysiological data in groups A and B were: MDP -80.74 +/- 1.94 mV and 44.54 +/- 6.24 mV, APA 92.72 +/- 9.25 mV and 57.74 +/- 10.85 mV, APD 50% 42.48 +/ 6.63 msec and 210.34 +/- 36.38 msec and APD 75% 56.47 +/- 8.55 msec and 281.66 +/- 42.18 msec respectively. The difference in the Na+-K+ ATPase activities between groups A (15.37 +/- 0.46 mumole Pi/mg/hr) and B (12.55 +/- 0.4 mumole Pi/mg/hr) was highly significant. CF molecules were frequently seen to be more irregularly and loosely distributed in the sarcolemmal surfaces of group B atrial myocytes. Based on these results we conclude that depression of the sarcolemmal Na+-K+ ATPase activity and derangement of the anionic binding sites in the sarcolemmal surfaces play an important role in altering transmembrane potentials in diseased human atria. PMID- 3027221 TI - Increased cyclic GMP in atrial fibrillation. AB - To investigate the role of cyclic nucleotides in the genesis and/or the persistence of atrial fibrillation (AF), plasma levels of cyclic GMP (c-GMP) and cyclic AMP (c-AMP) were measured in dogs with electrically induced AF, and in dogs subjected to high frequency (230 or 410/min) atrial pacing. The atrial fibrillation threshold (AFT) after intravenous administration of a dibutyryl derivative of c-GMP (Dbc-GMP), and the effect of atropine (0.1 mg/kg) on AFT were determined. Plasma levels of c-GMP and c-AMP were also measured in patients during paroxysmal AF and sinus rhythm. The c-GMP level increased significantly 15 min after the onset of artificial AF, and gradually increased during the course of the experiment. The c-GMP level began to increase significantly 15 min after the initiation of pacing at the higher rate (410/min) but not at the lower rate, compared to the prepacing value. The c-GMP level continued to rise until the end of pacing in the animals paced at 410/min. Although Dbc-GMP induced a dose dependent decrease in AFT, atropine did not prevent the decrease in AFT by Dbc GMP. In contrast, c-AMP levels were not significantly affected in any of these experiments. Clinical assessment revealed that patients had almost four times higher c-GMP level during AF than during sinus rhythm, though c-AMP levels in these patients did not change significantly during the attack of AF. These results suggest that the increase in c-GMP plays an important role in the maintenance and/or the genesis of AF. PMID- 3027222 TI - Cytochemical quantitation of cytochrome oxidase activity in rat pulmonary alveolar epithelial cells and possible defect in type I cells. PMID- 3027223 TI - High voltage electron microscopy of whole cells infected with herpes simplex virus type 1. PMID- 3027224 TI - Release of aldosterone and corticosterone from the adrenal cortex of the Brattleboro rat in response to administration of ACTH. AB - The time-course and dose-response of the in-vivo secretion of aldosterone and corticosterone after administration of ACTH(1-24) were measured in adrenal venous blood from female Brattleboro rats, homozygous for hypothalamic diabetes insipidus and lacking arginine vasopressin (AVP). Female Long-Evans rats were used as controls. All animals were pretreated with dexamethasone and anaesthetized with pentobarbital. Basal secretions of aldosterone and corticosterone were four- to sixfold lower in Brattleboro than in Long-Evans rats. Administration of ACTH consistently increased the secretion of aldosterone and corticosterone similarly in the two groups of rats; maximum values were observed 20-30 min after ACTH injection. However, for all the doses of ACTH (0.05, 0.5 and 5.0 mi.u./100 g body wt) and at every stage of response the secretion rates of aldosterone and corticosterone were twofold lower in Brattleboro than in Long-Evans rats. Furthermore the absolute increase in steroid secretion induced by ACTH was reduced by half in Brattleboro rats. These results show that the impairment of adrenal activity is largely due to a reduced capacity for corticosteroidogenesis in the adrenal cortex of Brattleboro rats. The mechanisms of action of AVP are discussed. PMID- 3027225 TI - Effects of forskolin on the morphology and function of the rat thyroid cell strain, FRTL-5: comparison with the effects of thyrotrophin. AB - The effect of diterpene forskolin, a potent stimulator of cyclic AMP (cAMP) in the rat thyroid cell strain FRTL-5, was compared with that of TSH. Forskolin stimulated both the release of cAMP into the culture medium and the accumulation of cAMP in the cytoplasm in a dose-dependent manner, within the range of 0.1-1000 mumol/l. Maximum cAMP concentrations were reached within 15 min of stimulation with forskolin. This is comparable with the effects of TSH. Forskolin also induced morphological changes in cultures of FRTL-5 cells, producing conspicuous cell retraction with arborization and numerous microvilli on the cell surface, specific reorganization of the microfilaments and modulation of the distribution of tubulin and fibronectin. Morphological changes induced by forskolin were always observed 20 to 30 min earlier, and in a higher percentage of cells, than the changes induced by TSH. Cell proliferation, however, was stimulated more effectively by TSH than by forskolin. These observations suggest that TSH might exert its effect on the morphology and growth of FRTL-5 cells, at least in part, through cAMP. The control of morphology and growth might not, however, be regulated solely by the adenylate cyclase and cAMP system. PMID- 3027226 TI - Prostaglandin F2 alpha-induced functional luteolysis: interactions of LH, prostaglandin F2 alpha and forskolin in cyclic AMP and progesterone synthesis in isolated rat luteal cells. AB - The acute antigonadotrophic action of prostaglandin F2 alpha (PGF2 alpha) was examined in dispersed luteal cell preparations from immature superluteinized rat ovaries. Cell suspensions prepared by collagenase digestion and purification over a Percoll density gradient were incubated for 1 h in Eagle's minimum essential medium in the presence and/or absence of LH, PGF2 alpha, N6,O2' dibutyryladenosine 3',5'-cyclic monophosphate (dbcAMP) and forskolin. Medium was assayed for total progesterone and adenosine 3',5'-cyclic monophosphate (cAMP). Luteal cell preparations showed typical steroidogenic (progesterone) responses to LH, mimicked by both dbcAMP and forskolin. Whilst the threshold LH dose to increase cAMP synthesis was greater than that for progesterone (100 micrograms/l compared with 1 microgram/l), 24 mumol forskolin/l was the threshold dose for both cAMP and progesterone responses. Furthermore, combined doses of LH and forskolin synergistically raised cAMP yet produced less than additive increases in progesterone. Similarly, combinations of dbcAMP plus forskolin produced less than additive progesterone increases. These data suggest that forskolin may not act as a simple mimic of LH. Prostaglandin F2 alpha dose-dependently inhibited forskolin-induced cAMP and progesterone synthesis and also inhibited progesterone synthesis induced by dbcAMP. These data suggest that the antigonadotrophic effect of PGF2 alpha has more than one locus of action, i.e. it both inhibits an adenylate cyclase event associated with cAMP generation and blunts the cellular response to cAMP. The present uncertainty over the exact locus of forskolin's action within the adenylate cyclase complex limits further delineation of the inhibitory action of PGF2 alpha on LH-responsive adenylate cyclase. PMID- 3027227 TI - Met-enkephalin stimulates secretion of ACTH in sheep. AB - The ability of opioid peptides to affect plasma immunoreactive ACTH concentrations was examined in conscious sheep. Plasma concentrations of ACTH were significantly increased by an i.v. infusion of Met-enkephalin and its analogue (FK-33824). There was a dose-dependent increase in plasma concentrations of ACTH with a graded administration of FK-33824. The combined effect of Met enkephalin and ovine corticotrophin-releasing factor (oCRF) or FK-33824 and oCRF on plasma concentrations of ACTH was greater than the effect of each peptide when given individually. This study demonstrates that Met-enkephalin and its analogue stimulate ACTH release in sheep. PMID- 3027228 TI - Effect of long-term infusion of ovine corticotrophin-releasing factor in the immature ovine fetus. AB - Synthetic ovine corticotrophin-releasing factor (oCRF) was infused continuously into the jugular veins of six ovine fetuses for 5-11 days. Two fetuses receiving 0.1 and 1.0 microgram oCRF/h from gestational days 134 and 135 respectively, lambed prematurely on days 141 and 140 respectively. Three out of four fetuses receiving oCRF at 2.4 micrograms/h, from 125 days of gestation, delivered spontaneously at 131, 131 and 136 days, whilst one died in utero at 132 days. Two fetuses receiving vehicle only or oCRF intra-amniotically, were born at 148 and 145 days respectively, whilst six fetuses chronically cannulated but not infused were born at 149.8 +/- 2.1 (S.D.) days. In ewes lambing at term, maternal plasma progesterone concentrations were 41.4 +/- 11.4 (S.E.M.; n = 5), 28.8 +/- 7.8 (n = 6), 17.1 +/- 4.8 (n = 5) and 7.9 +/- 1.1 (n = 4) nmol/l on 3, 2, 1 and 0 days respectively before the lambs were born. No such decrease in maternal plasma progesterone concentrations was seen in the oCRF-infused fetuses. Fetal plasma concentrations of immunoreactive ACTH were maintained above normal in oCRF infused fetuses, but some desensitization to bolus oCRF injections occurred in these fetuses. Four of the five fetuses born prematurely were sufficiently mature to survive, being able to stand, breathe and suckle. It is concluded that continuous oCRF infusions into immature fetuses can accelerate maturation of a number of organs and systems culminating in the premature delivery of viable lambs. PMID- 3027229 TI - Interference in the cytochemical assay of plasma vasopressin by a circulating factor induced by salt-loading in man. AB - Infusion of hypertonic saline into six normal volunteers caused an increase in plasma osmolality from 286.8 +/- 1.7 (mean +/- S.E.M.) to 307.6 +/- 2.6 mosmol/kg (P less than 0.001), a 7.1% increase in estimated blood volume, a rise in plasma immunoreactive arginine vasopressin (AVP) concentrations from 1.3 +/- 0.2 to 12.7 +/- 3.6 pmol/l (P less than 0.001) but no change in plasma AVP concentrations (2.1 +/- 0.9 and 1.9 +/- 1.3 pmol/l) as measured by a cytochemical technique based on the ability of AVP to stimulate rat renal medullary Na+/K+-ATPase activity. Addition of synthetic AVP to plasma obtained before, during and after hypertonic saline infusion also failed to stimulate Na+/K+-ATPase activity. The results suggest that infusion of hypertonic saline interfered with the cytochemical assay for AVP by inhibiting AVP-stimulated medullary Na+/K+-ATPase activity. We conclude that the use of this cytochemical method to detect plasma AVP has severe limitations under these experimental conditions. PMID- 3027230 TI - Arginine infusion blocks the action of parathyroid hormone but not arginine vasopressin on the renal tubule in man. AB - Six male volunteers were infused with arginine (0.5 g/kg body weight) over 30 min, after an overnight fast and water deprivation. There was a significant decrease in renal phosphate clearance (P less than 0.025) and urinary cyclic adenosine monophosphate (cAMP) output (P less than 0.025) during the 60- to 90 min period after the beginning of the infusion; both returned to the preinfusion basal levels within 150 min. The plasma levels of parathyroid hormone (PTH) were not affected by the infusion and remained unchanged during the subsequent 150 min. Plasma levels of arginine vasopressin (AVP) were also not significantly affected although plasma osmolality increased by 6-9 mmol/kg in all subjects. The infusion resulted in a diuresis, and a fall in urine osmolality but a decrease in free-water clearance; creatinine clearance was not affected. Six other subjects were given a bolus of 230 i.u. PTH intravenously, and 20 days later this was repeated during an infusion of arginine (0.5 g/kg body weight). There was a significant decrease in urinary phosphate (P less than 0.025) and cAMP excretion (P less than 0.05) when PTH was given with arginine. It is suggested that arginine blocks the action of PTH on the proximal renal tubule but not that of vasopressin on the distal nephron and collecting ducts. PMID- 3027231 TI - Genetic activity at the albino locus in Cattanach's insertion in the mouse. AB - Cattanach's insertion (Is(In7;X)1Ct or XCt) includes the normal allele at the albino locus (c+), which is subject to inactivation of the X chromosome carrying it, so that XCtX; cc mice have albino and pigmented patches. The X-autosome translocation T(X;16)16H or XT16H leads to preferential inactivation of the other X chromosome in female cells, so that XCtXT16H; cc mice are almost entirely white. However, they grow darker with age, as if reversal of inactivation of the c+ allele were taking place in increasing numbers of melanocytes. To test whether this is dependent only on age or whether it is related to the number of times the animal has moulted, hair was repeatedly plucked from selected areas at the early telogen stage when the follicles are also removed, assuming that the melanocytes or melanoblasts in that region of the skin would be forced to undergo further divisions to colonize the new follicles. The plucked areas grew darker at the same rate as the rest of the coat, suggesting that the progressive reversal of inactivation is dependent only on age. As direct examination of melanocytes in the follicles is difficult, they were examined in the choroid and the retinal pigment epithelium (RPE) of the eye. The frequency of the pigmented cells was lower in the choroid than in the RPE. Since the melanocytes in these structures are different in origin as well as in physical characteristics, it appears that cell type influences either reversal of inactivation, or the frequency with which the influence of the X chromosome extends to the albino locus. PMID- 3027232 TI - Scanning electron microscopic observation of mouse embryonic submandibular glands during initial branching: preferential localization of fibrillar structures at the mesenchymal ridges participating in cleft formation. AB - Branching submandibular glands of 12-day mouse embryos and those cultured in the presence and absence of a collagenase inhibitor from the culture medium of bovine dental pulp or a Clostridial collagenase were examined with the scanning electron microscope. Fracturing of fixed and dried glands with the tip of a fine needle succeeded in exposing the surfaces of the lobules and of their mesenchymal replicas at different stages of branching. At the beginning of branching, corresponding parts of the mesenchyme formed ridges on or in which the fibrillar structures were often found. At the stage forming deeper clefts thicker fibres, 0.5-2.5 micron in diameter, were observed between two adjacent lobules. On the contrary, no apparent differences in the fibrillar structures on the epithelial surfaces were detected between the shallow cleft and noncleft regions at the initial phase of branching. These fibrillar structures were very abundant in glands cultured with collagenase inhibitor and were completely lost in glands cultured with bacterial collagenase, strongly indicating that these materials consisted of collagen. The possible involvement of mesenchyme in epithelial branching is discussed with special reference to mesenchymal traction forces that would be elicited by fibrillar collagens. PMID- 3027234 TI - Some properties of a Saccharomyces cerevisiae mutant resistant to 2-amino-4 methyl-5-beta-hydroxyethylthiazole. AB - A mutant of Saccharomyces cerevisiae highly resistant to 2-amino-4-methyl-5-beta hydroxyethylthiazole (2-aminohydroxyethylthiazole), an antimetabolite of 4-methyl 5-beta-hydroxyethylthiazole (hydroxyethylthiazole), has been isolated. Its resistance to 2-aminohydroxyethylthiazole was about 10(4) times that of the sensitive parent strain. The amount of thiamin synthesized in the cells of the resistant strain grown in minimal medium was less than half of that of the sensitive strain. The ability to synthesize thiamin from 2-methyl-4-amino-5 hydroxymethylpyrimidine (hydroxymethylpyrimidine) and hydroxyethylthiazole in the resistant strain was low compared with that of the sensitive strain. These results were found to be due to a deficiency of hydroxyethylthiazole kinase in the resistant strain: in sonic extracts of cells the enzyme activity was only 0.67% of that of the sensitive strain. Although the cells of the sensitive strain could accumulate exogenous hydroxyethylthiazole in the form of hydroxyethylthiazole monophosphate, no significant uptake of hydroxyethylthiazole by the cells of the resistant strain was observed. The possibilities that 2 aminohydroxyethylthiazole monophosphate may be the actual inhibitor of the growth of Saccharomyces cerevisiae, and that hydroxyethylthiazole may not be involved in the pathway of de novo synthesis of thiamin via hydroxyethylthiazole monophosphate, are discussed. PMID- 3027233 TI - Acid-base regulation and ion transfers in the carp (Cyprinus carpio): pH compensation during graded long- and short-term environmental hypercapnia, and the effect of bicarbonate infusion. AB - To study both temporal and quantitative effects of hypercapnia on the extent of pH compensation in the arterial blood, specimens of carp (Cyprinus carpio) were exposed to a PCO2 of about 7.5 mmHg (1 mmHg = 133.3 Pa) (1% CO2) in the environmental water for several weeks, and a second group of animals was subjected to an environmental PCO2 of about 37 mmHg (5% CO2) for up to 96 h. A third series of experiments was designed to test the possibility that infusion of bicarbonate would increase the extent of plasma pH compensation. Dorsal aortic plasma pH, PCO2 and [HCO3-], as well as net transfer of HCO3- -equivalent ions, NH4+, Cl- and Na+, between fish and ambient water, were monitored throughout the experiments. Exposure to environmental PCO2 of 7.5 mmHg resulted in the expected respiratory acidosis with the associated drop in plasma pH, and subsequent compensatory plasma [HCO3-] increase. The compensatory increase of plasma bicarbonate during long-term hypercapnia continued during 19 days of exposure with plasma bicarbonate finally elevated from 13.0 mmoll-1 during control conditions to 25.9 mmoll-1 in hypercapnia, an increase equivalent to 80% plasma pH compensation. Exposure to 5% hypercapnia elicited much larger acid-base effects, which were compensated to a much lesser extent. Plasma pH recovered to only about 45% of the pH depression expected at constant bicarbonate concentration. At the end of the 96-h exposure period, plasma [HCO3-] was elevated by a factor of 2.5 to about 28.2 mmoll-1. The observed increase in plasma bicarbonate concentration during 5% hypercapnic exposure was attributable to net gain of bicarbonate equivalent ions from (or release of H+-equivalent ions to) the environmental water. Quantitatively, the gain of 15.6 mmol kg-1 was considerably larger than the amount required for compensation of the extracellular space, suggesting that acid-base relevant ions were transferred for compensation of the intracellular body compartments. The uptake of bicarbonate equivalent ions from the water was accompanied by a net release of Cl-and, to a smaller extent, by a net uptake of Na+, suggesting a 75% contribution of the Cl /HCO-3 exchange mechanism. Infusion of bicarbonate after 48 h of exposure to 7.5 mmHg PCo2 had only a transient effect on further pH compensation. The infused bicarbonate was lost to the ambient water, and pre-infusion levels of bicarbonate were reattained within 24 h. Repetition of the infusion did not result in a notable improvement of the acid-base status.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3027235 TI - Expression of the Aspergillus nidulans argB gene in Escherichia coli. AB - The Aspergillus nidulans argB gene coding for ornithine carbamoyltransferase (OTCase) is not expressed in Escherichia coli. However, E. coli OTCase-deficient strains transformed with plasmids carrying the argB gene from A. nidulans reverted to prototrophy at a high frequency. In these derivatives the argB gene became functional due to DNA rearrangements upstream of the coding sequence. Two types of rearrangement were characterized. One was identified as an insertion of IS2. The second was a deletion that resulted in transcription of the argB gene from the TcR gene promoter and translation from a newly created ribosome-binding site formed at the junction between the A. nidulans and vector DNA sequences. PMID- 3027236 TI - The expression of the Escherichia coli lacZ gene in Streptomyces. AB - The Escherichia coli lacZ gene was stably introduced into phi C31-based phage cloning vectors in Streptomyces lividans. However, lacZ could not be stably introduced into S. lividans on the plasmid vectors pIJ702 or pIJ41. Studies of the expression of lacZ in S. lividans and S. coelicolor were facilitated by the use of mutants and/or growth conditions in which endogenous beta-galactosidase activity was low or absent. Plaques and lysogens of phi C31::lacZ constructs involving transcriptional fusions to the pBR322 tet gene contained beta galactosidase activity. Activities were markedly higher in S. lividans than in S. coelicolor. Insertion of the major transcriptional terminator of coliphage fd between the tet promoter and lacZ reduced lacZ expression more in lysogens than in lytically infected cultures, suggesting the existence of a phage-specified anti-termination function. Several phi C31 derivatives suitable for the construction of different kinds of transcriptional and translational fusions in Streptomyces were derived. The expression of lacZ in one transcriptional fusion vector (KC659) was increased both in plaques and in lysogens by the appropriately positioned insertion of a previously characterized promoter from the aph gene of S. fradiae. When phi C31 DNA fragments were inserted into KC659, at least one recombinant phage gave high beta-galactosidase activity in lytic infections, but not in lysogens. The potential usefulness of lacZ fusions in Streptomyces is discussed. PMID- 3027237 TI - Three mutants of herpes simplex virus type 2: one lacking the genes US10, US11 and US12 and two in which Rs has been extended by 6 kb to 0.91 map units with loss of Us sequences between 0.94 and the Us/TRs junction. AB - In the process of generating restriction endonuclease site deletion mutants, we have isolated and characterized three mutants of herpes simplex virus type 2 (HSV 2), strain HG52, with large genomic deletions in Us and TRs. The deleted sequences (7.5 kb) extend from 0.94 map coordinates (m.c.) to 0.99 m.c. and are presumed to contain the HSV-2 gene equivalents of US10, 11 and 12, one copy of immediate early (IE) gene 3 and one copy of an origin of replication (ORIs). One of the mutants (HG52X163X12) has a simple deletion whereas in the two others (HG52X163X14 and HG52X163X21) the deleted sequences have been replaced by inverted duplications of Us/IRs sequences between 0.83 and 0.91 m.c. such that the molecules have short region inverted repeats extended by 6 kb on either side. All three are viable, stable and grow in tissue culture indicating that the polypeptides coded by the HSV-2 genes equivalent to US10, 11 and 12 are non essential for lytic growth in BHK21/C13 cells. In addition the lack of one copy of the HSV-2 equivalent of IE gene 3 and ORIs in HG52X163X12 shows that only one copy of each suffices for viability. However the temperature restriction of the mutants at 38.5 degrees C suggests that one or more of the polypeptides coded by the deleted sequences may be required in conjunction with another polypeptide(s) for viral growth or stability at the higher temperature. PMID- 3027238 TI - Isolation and preliminary characterization of Semliki Forest virus mutants with altered pathogenicity for mouse embryos. AB - Four temperature-sensitive (ts) mutants of the A7 strain of Semliki Forest virus (SFV) have been isolated. All mutants were defective in RNA synthesis at the restrictive temperature (39 degrees C) compared to the permissive temperature (30 degrees C). Since the body temperature of mice fluctuates between 37 degrees C and 39 degrees C, multiplication was also examined at 37 degrees C; only the multiplication of ts4 was restricted. After intraperitoneal infection of 8-day pregnant mice, the wild-type induced rapid abortion. Ts4 and ts26 had no effect on embryonic development. Litters born to ts4-infected mothers developed no postnatal immunity whereas 50% of litters from ts26-infected mothers were immune. Unlike the wild-type, ts14 induced the same or higher virus titres in placental tissue in most mice than in foetal tissue. Ts22 and ts14 induced a range of development defects, including developmental arrest, mummification, abortion and postnatal death. Most surviving offspring were immune. Although ts4 induced no viraemia, ts14, ts22 and ts26 induced a lower titre but longer lasting viraemia than the wild-type. It is concluded that infections of pregnant mice with ts14 and ts22 in particular are good models for analysis of the mechanism of virus induced developmental defects. PMID- 3027239 TI - Reassortment of human rotavirus possessing genome rearrangements with bovine rotavirus: evidence for host cell selection. AB - Mixed infections of secondary rhesus monkey kidney cells with human rotaviruses carrying rearranged genomes and with bovine rotavirus yielded a high percentage of reassortants. The genotypes of 511 plaque-purified clones raised in either MA104 or BSC-1 cells have been determined and the frequencies of different genotypes have been calculated. It was found that reassortants did not emerge at random; there was non-random association of certain genes; the cell line used to isolate reassortants influenced the result, i.e. host cell factors had a selective effect on a recombinational mixture. PMID- 3027240 TI - Biochemical evidence for the oligomeric arrangement of bovine rotavirus nucleocapsid protein and its possible significance in the immunogenicity of this protein. AB - The nucleocapsid protein of bovine rotavirus was shown to exist in trimeric units in both the virus particle and in infected cells, with the subunits linked by non covalent interactions. These trimeric units complex further by disulphide bridges into larger units which may represent the hexameric structures observed by electron microscopy. Visualization of various nucleocapsid protein complexes was also achieved on polyacrylamide gels by treating virus preparations with urea at 37 degrees C or boiling in the presence and absence of 2-mercaptoethanol. Since virus particles devoid of nucleic acid were also broken down into trimeric subunits by such treatments, assembly of virus particles appears not to require an RNA-protein interaction. Four nucleocapsid-specific monoclonal antibodies with low neutralizing ability reacted with the monomeric (45,000 mol. wt., 45K), dimeric (90K), trimeric (135K) and trimeric pair (270K) subunits, indicating that a site responsible for neutralization is probably exposed after assembly of these subunits. Analysis of radiolabelled virus revealed that a high proportion (80%) of infectious particles could be immunoprecipitated by these monoclonal antibodies, suggesting that the virus particles are either partially double shelled or have the nucleocapsid exposed on the surface. The monoclonal antibodies also cross-reacted with the nucleocapsid proteins of simian (SA11), pig (OSU), bovine (NCDV and UK) and human (Wa and ST4) rotaviruses in an immunoblot ELISA reaction. Since these six viruses belong to two different subgroups, it is likely that the antibodies did not recognize the subgroup specific site, but a shared exposed antigenic determinant. Due to the hexameric configuration of the nucleocapsid in virus particles the neutralizing epitope may be repeatedly presented and, therefore, may contribute to the immunogenicity of this protein. PMID- 3027241 TI - Bovine parvovirus DNA-binding proteins: identification by a combined DNA hybridization and immunodetection assay. AB - We have investigated the interaction between bovine parvovirus (BPV) capsid and non-capsid proteins and restriction fragments of the BPV genome by a combined DNA hybridization and immunodetection assay. 32P-labelled DNA was bound to nitrocellulose membranes bearing lysates of mock-infected and virus-infected cells whose proteins had been separated by SDS-polyacrylamide gel electrophoresis. The position of bound DNA was determined by autoradiography. The proteins on the membrane were still accessible to specific antibodies, allowing confirmation of the DNA-binding species by an immunodetection reaction. In 0.2 M NaCl, BPV capsid proteins VP2 (72,000 daltons) and VP3 (62,000 daltons) bound the 0 to 16 map unit EcoRI fragment of BPV DNA which contained label in either the minus or plus strand. At higher salt concentration (0.5 M), only VP2 still bound DNA. Within this fragment, the capsid protein binding was restricted to those nucleotides between map units 0 and 4. No binding to capsid proteins was seen with the fragment spanning the middle of the genome and minor binding to VP3 was seen with the 5' end. Binding to the BPV non-capsid protein NP-1 was observed with the 0 to 16 map unit fragment when label was in the virion strand and to other possibly BPV-coded proteins when label was in the plus strand. The NP-1 binding was localized to map units 4 to 16. We did not detect binding to the BPV homologue(s) of the autonomous parvovirus non-capsid protein NS1, due in part to its low concentration in the cell lysates used. Points of the parvovirus replication cycle at which DNA-binding proteins may serve controlling functions are discussed. PMID- 3027242 TI - DNA sequence and genetic content of the HindIII l region in the short unique component of the herpes simplex virus type 2 genome: identification of the gene encoding glycoprotein G, and evolutionary comparisons. AB - The DNA sequence was determined of the HindIII l fragment of herpes simplex virus type 2 (HSV-2), which is located in the short unique region of the HSV-2 genome. HindIII l was found to comprise 9629 base pairs. Comparison with the previously determined corresponding sequence for herpes simplex virus type 1 (HSV-1), and limited mRNA mapping, showed that HindIII l contained six genes (termed US2 to US7) and part of another (US8). The HSV-1 and HSV-2 sequences were found to be generally colinear, with one major exception: the HSV-2 DNA contained an extra sequence of about 1460 base pairs, in the coding region of gene US4. By use of an antiserum raised against an oligopeptide representing amino acids near the C terminus of the predicted HSV-2 US4 polypeptide it demonstrated that this gene encodes the virion glycoprotein gG-2, while HSV-1 US4 encodes a much smaller virion glycoprotein with homology to the C-terminal portion of gG-2. Quantitative comparisons of the HSV-2 HindIII l and corresponding HSV-1 sequences showed that they had diverged by point mutation and by local addition and deletion, as well as by the major change in genes US4. It was found that within the HSV-2-specific part of gG-2 there was a locality showing sequence similarity to a glycoprotein of pseudorabies virus (gX), and weaker similarity to glycoproteins D of HSV-1 and HSV-2. These data were interpreted to suggest, first, that HSV-2 US4 represents an ancient gene of alpha herpesviruses, and, more tentatively, that the evolution of the genes for gG and gD may have proceeded through a duplication event. PMID- 3027243 TI - Conserved homologous regions between two baculovirus DNAs. AB - Regions of homology on the physical maps of Spodoptera exempta multiple nucleocapsid nuclear polyhedrosis virus (SeMNPV-25), an Autographa californica MNPV genomic variant, and S. frugiperda (SfMNPV-2) baculovirus DNAs were identified by reciprocal DNA-DNA blot hybridization under conditions of an effective temperature of Tm -25 degrees C. In addition, cloned fragments of the viral genome which contained the homologous regions were used in hybridization experiments to confirm, refine and correlate the regions of the two physical maps. Five homologous regions conserved between the two physical maps were identified. When the stringency of the hybridization was increased (Tm -20 degrees C), only two of the original regions were identified by blot hybridization. One of the two regions contained the polyhedrin gene, and the other region was not associated with any known viral function. The five regions did not overlap with the intragenic homologous sequence (hr1 to hr5) regions on the SeMNPV-25 map or with restriction endonuclease variant (vI to vIV) regions on the SeMNPV-25 and SfMNPV-2 maps. The degree of similarity in the genomic organization of these two baculoviruses is discussed. PMID- 3027244 TI - Molecular cloning of the closed circular provirus of human T cell leukaemia virus type I: a new open reading frame in the gag-pol region. AB - A DNA clone of human T cell leukaemia virus type I (HTLV-I) was isolated from extrachromosomal closed circular copies in chronically infected promyelocytic leukaemia HL60 cells. The new HTLV-I isolate had an intact reading frame in the gag-pol region which could encode protein of 234 amino acids. This open reading frame has not been observed in previous HTLV-I isolates, although similar open reading frames have been reported in the corresponding locations in the related bovine leukaemia virus and HTLV type II. We consider that this open reading frame codes for the virus-encoded protease, on the basis of the homology of the predicted amino acid sequence with those of previously identified retrovirus proteases. PMID- 3027245 TI - Simian virus 40-induced mutagenesis: action of the early viral region. AB - Earlier results have demonstrated a mutagenic activity of simian virus 40 (SV40) in mammalian cells. To analyse this ability further, the effect of SV40 DNA fragments, introduced into Chinese hamster cells, on the frequency of mutations at the hypoxanthine phosphoribosyltransferase locus and other loci was studied. It was found that the mutagenic effect was substantially maintained when the viral genome had been replaced by a fragment comprising the T antigen-coding region and the early promoter-enhancer region; was strongly reduced or abolished when the promoter region including upstream sequences in this fragment had been replaced by the chicken lysozyme gene promoter or both enhancer elements were deleted, and was abolished in an SV40 replication origin-defective mutant in which the structure of the T antigen-binding site II was affected. It may be concluded that SV40-induced mutagenesis depends on the expression of the early region of the genome and on a function involved in specific binding of large T antigen to viral DNA. Since origin-defective mutants of SV40 were reported as being able to transform cells, the functions of transformation and mutation do not seem to correlate. PMID- 3027246 TI - Stability of a bacterial gene in a bovine papillomavirus-based shuttle vector maintained extrachromosomally in mammalian cells. AB - In order to analyse the stability of cloned genes in a viral vector we have constructed a shuttle vector based on bovine papillomavirus and the Escherichia coli gene lacZ. Propagation of this vector in mouse C127 cells and analysis of vector sequences in bacteria produced no detectable mutations in the lacZ gene in over 6137 clones analysed. This is 100-fold less than the mutation frequency observed when the same and similar target genes are replicated in monkey COS cells using a simian virus 40-based shuttle vector. PMID- 3027247 TI - Analysis of the role of the cysteine 171 residue in the activity of herpes simplex virus type 1 thymidine kinase by oligonucleotide-directed mutagenesis. AB - The thymidine kinase (TK) gene from herpes simplex virus type 1 strain SC16 was cloned into bacteriophage M13 mp8 so that functional HSV-1 TK was expressed in bacteria infected with the recombinant bacteriophage, M13/TK. Oligonucleotide site-directed mutagenesis was then employed to introduce single nucleotide changes into the TK gene in M13/TK in order to alter the codon for cysteine 171 in the wild-type enzyme to a codon specifying either serine or glycine. Analysis of the mutant enzymes in bacterial extracts showed that these substitutions had little effect on the activity of the enzyme, indicating that the side chain of this residue is not involved in nucleoside binding and is not essential for the catalytic activity of the enzyme. PMID- 3027248 TI - Nucleotide sequence of the gene encoding the surface projection glycoprotein of coronavirus MHV-JHM. AB - Sequences encoding the surface projection glycoprotein of the coronavirus, murine hepatitis virus (MHV), strain JHM, have been cloned into pAT153 using cDNA produced by priming with specific oligonucleotides on infected cell RNA. The regions of three clones pJMS1010, pJS112 and pJS92, which together encompass the surface protein gene have been sequenced by the chain termination method. The sequence of the primary translation product, deduced from the DNA sequence, predicts a polypeptide of 1,235 amino acids with a molecular weight of 136,600. This polypeptide displays the features characteristic of a group 1 membrane protein; an amino-terminal signal sequence and carboxy-terminal membrane and cytoplasmic domains. There are 21 potential glycosylation sites in the polypeptide and a cysteine-rich region in the vicinity of the transmembrane domain. During maturation proteolytic processing of the polypeptide occurs and at positions 624 to 628 the sequence Arg-Arg-Ala-Arg-Arg is found, which is similar to a number of basic sequences involved in the cleavage of enveloped RNA virus glycoproteins. The fusogenic properties of the MHV surface protein do not appear to correlate with a strongly hydrophobic region at the putative amino terminus of the carboxy-terminal cleavage product. PMID- 3027249 TI - Completion of the sequence of the genome of the coronavirus avian infectious bronchitis virus. AB - The nucleotide sequence determination of the genome of the Beaudette strain of the coronavirus avian infectious bronchitis virus (IBV) has been completed. The complete sequence has been obtained from 17 overlapping cDNA clones, the 5'-most of which contains the leader sequence (as determined by direct sequencing of the genome) and the 3'-most of which contains the poly(A) tail. Approximately 8 kilobases at the 3' end of this sequence have already been published. These contain the sequences of mRNAs A to E within which are the genes for the spike, the membrane and the nucleocapsid polypeptides: the main structural components of the virion. The remainder of the sequence, equivalent to the 'unique' region of mRNA F, is some 20 kilobases in length and is thought to code for a polymerase or polymerases which are involved in the replication of the genome and the production of the subgenomic messenger RNAs. This sequence contains two large open reading frames, potentially coding for polypeptides of molecular weights 441,000 and 300,000. Unlike other large open reading frames in the virus, the 300,000 open reading frame appears to have no subgenomic RNA associated with it which would allow it to be at the 5' end of an mRNA species. Because of this, and because of the characteristics of the sequence in the region immediately upstream of its start codon, other mechanisms of translation, such as ribosome slippage, must be postulated. PMID- 3027250 TI - Dose pipecolic acid interact with the central GABA-ergic system? AB - Several previous studies have suggested a strong GABA-mimetic action of the endogenous brain imino acid, L-pipecolic acid (L-PA). In the present study, these observations were evaluated using electrophysiological and neurochemical methods. In contrast to published data our electrophysiological studies on rat cortical neurones in situ showed only a weak, but bicuculline-sensitive depressant action of L-PA on cortical neurones. Furthermore, L-PA proved to have no affinity for any of the three components of the GABA-benzodiazepine-chloride channel receptor complex. However, using a modification of published methods a weak affinity for the GABA-B receptor site was demonstrated (IC50 = 1.8 X 10(-3) M). L-PA showed no anticonvulsive activity in several tests; in particular, it did not protect mice from seizures induced by inhibition of L-glutamate-1-decarboxylase (EC 4.1.1.15: GAD). L-PA had a very weak action on brain GABA levels of mice, and did not modify the rate of GABA synthesis. In conclusion, these results are not compatible with a strong in vivo interaction between L-PA and GABA-mediated inhibitory transmission. PMID- 3027251 TI - Chronic beta-adrenergic stimulation increases in mice the sensitivity to methysergide and the number of cerebral high affinity serotonin binding sites (5 HT-1). AB - Reserpine administration in mice causes, among other effects an akinesia which can be reversed by the serotonin agonist-antagonist methysergide. The effect of methysergide is potentiated by clenbuterol, a beta-adrenergic agonist, which itself causes hypomotility. Potentiation is weak after a single injection of clenbuterol, but becomes much stronger after repeated administration for 12 days. This treatment also causes a 50% increase in the number of high affinity 5-HT-1 binding sites in the brain. This increase would explain the increased potency of methysergide against reserpine-induced akinesia. These results show that: a beta adrenergic drug, clenbuterol modulates the serotoninergic system; this modulation takes its importance after chronic treatment only; this interrelation may be important in depressive illness since it is observed on a test used in the screening of antidepressant drugs. PMID- 3027252 TI - EPR characterization of alcohol complexes of ferric myoglobin and hemoglobin. AB - Frozen solution electron paramagnetic resonance spectra of the aquo, methanol, and ethanol complexes of ferric myoglobin and hemoglobin are quantitatively analyzed in terms of the rhombic to tetragonal symmetry ratio and the admixture of quartet states, both with regard to central values of these parameters and the widths of their distributions. In both the methanol and ethanol complexes of ferric myoglobin the main change from the aquo complex is a narrowing of the spread in the rhombic to tetragonal symmetry ratio (reduction in structural variation). The alcohol complexes of both the alpha- and beta-chains within the tetramer of ferric hemoglobin are characterized by a lowering of symmetry (as compared with the aquo complex). Qualitative differences in distribution widths among the complexes are consistent with an origin in molecular structure and dynamics rather than in ice matrix-induced strain. PMID- 3027253 TI - Metalloenzymes as molecular switches: the role of conformation changes in controlling activity. AB - Enzymatically important conformation changes have been classified according to the nature of the trigger (capital S, I, A for substrate, reaction intermediate, allosteric modifier) and the rate of change (subscript f or s, if fast or slow relative to the enzymatic reaction). It is suggested that, in Cu superoxide dismutase, the known protonation of the product (to give H2O2) under nonequilibrium conditions involves an essentially irreversible If change. PMID- 3027254 TI - Determination of the optical properties of CuA(II) in bovine cytochrome c oxidase using magnetic circular dichroism as an optical detector of paramagnetic resonance. AB - This paper describes the use of a novel magnetic circular dichroism-microwave double resonance (MCD-ODMR) experiment to study the optical properties of the EPR detectable copper center, CuA2+, in bovine cytochrome c oxidase. By irradiating the sample with a monochromatic microwave beam of appropriate frequency it is possible to quench partially the MCD intensity of the features due to CuA2+. In this way the MCD bands from this center have been identified even in the presence of overlapping optical transitions from the haem centers of this enzyme. The resulting spectrum compares well with that reported some years ago from this laboratory and obtained by measuring MCD magnetisation characteristics. In addition the shapes of the MCD-ODMR lines obtained in a plot of MCD intensity against magnetic field strength have been analyzed to yield the relative polarizations of the optical transitions of CuA2+ which contribute to the MCD spectrum. All of the bands observed between 450 and 850 nm are predominantly polarized in the xy plane perpendicular to the direction of the g-tensor component of CuA2+ at g parallel = 2.18. This suggests that all of the CuA2+ ligands that contribute to the optical charge-transfer transitions in the visible region lie approximately in the basal plane. Possible structures for CuA2+ can now be suggested. PMID- 3027255 TI - Effects of synergistic anions on the proton magnetic resonance spectra and pH titration of histidinyl side chains in the C-terminal half-molecule of ovotransferrin. AB - High-resolution proton magnetic resonance spectra of the C-terminal half-molecule of ovotransferrin (OTf/2C) clearly resolve the C(2)H resonances of the five histidinyl residues in the protein. Formation of the Ga(III)OTf/2C(anion) ternary complexes results in different chemical shift and titration behaviors for certain C(2)H resonances in the carbonato, oxalato, and malonato complexes. The pKa' of the imidazole group involved in a proton relay with the synergistic anion and a water of hydration appears to be anion independent. Thus the initial attack of a proton on the ternary complex appears to be at a ligand other than the anion or anion-binding imidazole group. PMID- 3027256 TI - On the rates of enzymatic, protein and model compound reactions: the importance of diffusion control. AB - A meaningful method of comparison is suggested for saturation kinetics, typical of enzyme-catalyzed reactions, and nonsaturation kinetics, often typical of model compound reactions. True diffusion-controlled reactions do not give saturation behavior; but enzymes may need saturation behavior to attain selectivity and stereospecificity for complicated substrates or for reactions beyond the complexity of electron transfer. However, the diffusion controlled limit provides a better reference point for rate comparisons than does the rate of uncatalyzed reaction. The failure of the Stokes-Einstein equation for small substrates is documented, as are ways of circumventing the problem. Advantages and pitfalls in the use of viscosogens to test for diffusion control are delineated. Finally, the possible advantages of surface diffusion for an enzyme, but lack of experimental evidence, is discussed. PMID- 3027257 TI - Proton NMR studies of two helices in tuna ferricytochrome c. AB - 1H Nuclear magnetic resonance assignments are given for the NH and C alpha H protons of two alpha-helical segments of tuna ferricytochrome c. The assignments were obtained using two-dimensional nuclear magnetic resonance sequential assignment procedures and illustrate the applicability of these methods to medium sized proteins. By comparing nuclear Overhauser intensities between the NH and C alpha H protons the precise structures of the two helical segments are compared and their deviations from ideality are discussed. PMID- 3027258 TI - gamma-Aminobutyric acid receptor ionophore complexes: differential effects of deltamethrin, dichlorodiphenyltrichloroethane, and some novel insecticides in a rat brain membrane preparation. AB - The binding of [35S]t-butylbicyclophosphorothionate ([35S]TBPS), a gamma aminobutyric acid (GABA)-activated chloride ionophore ligand; [3H]diazepam, a benzodiazepine agonist; and [3H]muscimol, a GABA receptor probe, were used to assess the effects at 100 microM of deltamethrin, dichlorodiphenyltrichloroethane (DDT), and three experimental insecticides--a DDT-pyrethroid hybrid, GH414 (cycloprothrin), and two DDT-analogues, GH266 and GH149 (EDO), on GABA receptor ionophore complexes in a rat brain membrane preparation. GH266 and GH149 were found to inhibit a greater percentage of [35S]TBPS binding than the same concentration of deltamethrin or DDT, although GH414 had little effect. GH266 and GH149 enhanced [3H]diazepam binding by nearly 200%, in contrast to the inhibitory effects of deltamethrin, DDT, and GH414. GH266 and GH149 also caused a dramatic enhancement of [3H]muscimol binding, 367 and 236% of control, respectively, whereas DDT and deltamethrin caused only a moderate enhancement. The effects of the insecticides on binding affinity and density were examined for each of the ligands. The results show a differential interaction of the insecticides on the various ligand binding sites. PMID- 3027259 TI - 31P nuclear magnetic resonance in vivo spectroscopy of the metabolic changes induced in the awake rat brain during KCN intoxication and its reversal by hydroxocobalamine. AB - Radiofrequency surface coils were chronically implanted in rats, which were subsequently subjected to 31P nuclear magnetic resonance (NMR) investigations at 4.7 T. The implanted coil allowed study of the animals without need for anesthesia, which is a prerequisite for studies of normal brain metabolism. The animals may be kept in the NMR probe for several hours. During subsequent experiments, they may be placed in the same position, therefore allowing follow up studies for periods as long as 2 months. This method has been used in the study of sublethal KCN intoxication. KCN, a cytochrome c oxidase inhibitor, induces a blockade of cell respiratory processes, which is reflected, in a dose dependent manner, by a decrease in phosphocreatine content and pH and an increase in inorganic phosphate content, whereas ATP levels remain constant until high doses of KCN (6 mg/kg i.p.) are reached. 31P NMR allows the time course of these metabolic changes to be followed. For high KCN doses, a new peak, termed X, is observed, which is interpreted as being due to a pool of inorganic phosphate at very low pH (5.65), corresponding to a subset of cells that did not survive KCN injury. Hydroxocobalamine, a specific antidote of KCN, suppresses the metabolic changes due to 6 mg/kg of KCN. PMID- 3027260 TI - Targeting of liposomes to cells bearing nerve growth factor receptors mediated by biotinylated nerve growth factor. AB - We have used biologically active derivatives of beta-nerve growth factor (NGF), modified by biotinylation via carboxyl groups, to target the specific binding of liposomes to cultured rat and human tumor cells bearing NGF receptors. Liposomes, to be used for targeting, were prepared by conjugating streptavidin to phospholipid amino groups on liposomes prepared by reverse-phase evaporation. Approximately 2,000 streptavidin molecules were incorporated per liposome. Addition of biotinylated NGF, but not of unmodified NGF, could mediate the subsequent binding of radiolabeled streptavidin-liposomes to rat pheochromocytoma PC12 cells in suspension at 4 degrees C. In contrast, incubation with biotinylated NGF did not mediate the binding of hemoglobin-conjugated liposomes. Under optimal incubation conditions, approximately 570 streptavidin-liposomes were specifically bound per cell. Biotinylated NGF was also used to obtain specific binding of streptavidin-liposomes containing encapsulated fluorescein isothiocyanate-labeled dextran to PC12 cells or human melanoma HS294 cells. When HS294 cells were incubated at 37 degrees C following targeted liposome binding at 4 degrees C, the cell-associated fluorescence appeared to become internalized, displaying a perinuclear pattern of fluorescence similar to that observed when lysosomes were stained with acridine orange. Trypsin treatment abolished cell associated fluorescence when cells were held at 4 degrees C but did not alter the fluorescence pattern in cells following incubation at 37 degrees C. When liposomes containing carboxyfluorescein, a dye capable of diffusing out of acidic compartments, were targeted to HS294 cells, subsequent incubation at 37 degrees C resulted in diffuse cytoplasmic fluorescence, suggesting that internalized liposomes encounter lysosomal or prelysosomal organelles. PMID- 3027261 TI - Neuromelanogenic and cytotoxic properties of canine brainstem peroxidase. AB - We have isolated a heme protein from canine midbrains that possesses potent peroxidase activity. This enzyme catalyzes the oxidation of dopamine to neuromelanin in the presence of H2O2. We have further shown that the isolated peroxidase possesses potent cytotoxic activity in the presence of superoxide or H2O2 and Cl-. The enzyme possesses an endogenous NAD(P)H oxidase activity that can promote the cytotoxic activity by virtue of its production of superoxide. Other enzymes such as dihydroorotate dehydrogenase and galactose oxidase, which produce O2- and H2O2, respectively, are also effective in promoting the cytotoxic activity of the brainstem peroxidase. Although rat erythrocytes were routinely used as the target cell, other cell types, including rat hepatoma and mouse neuroblastoma cells, are also susceptible to the toxic action of the peroxidase. The cytotoxic action of the brainstem peroxidase is dramatically enhanced by kainic acid and is significantly enhanced by Mn2+, whereas dopamine was found to be a potent inhibitor of the cytotoxic activity. Based on these findings, we postulate a central role for the brainstem peroxidase in dopamine metabolism as well as in the biochemical and anatomical changes associated with Parkinson's disease. PMID- 3027263 TI - Effects of cerebral ischemia on [3H]inositol lipids and [3H]inositol phosphates of gerbil brain and subcellular fractions. AB - Intracerebral injection of [3H]inositol into gerbil brain resulted in labeling of phosphoinositides and inositol-phosphates in various subcellular membrane fractions. Phosphatidylinositol (PI) comprised greater than 90% of the radioactivity of inositol lipids. However, the level of labeled poly-PI (with respect to PI) was higher in synaptosomes than in other membrane fractions. Ischemia induced in gerbils by ligation of the common carotid arteries resulted in a 30% decrease in labeled poly-PI in brain homogenates and this decrease was largely attributed to the poly-PI in synaptosomes (50% decrease). Among the inositol phosphates, the ischemia induction resulted in a decrease in labeling of inositol triphosphate (63%) and inositol biphosphate (38%), but labeling of inositol phosphate (IP) was increased by 59%. The results suggested a rapid turnover of the inositol phosphates in the gerbil brain. In general, changes in inositol lipids and inositol phosphates due to ischemia were attenuated after pretreatment with lithium (3 meq/kg) injected intraperitoneally 5 h prior to ligation. Surprisingly, lithium treatment alone did not cause an increase in IP labeling in the gerbil brain. PMID- 3027262 TI - Purification and characterization of a peptidyl dipeptidase resembling angiotensin converting enzyme from the electric organ of Torpedo marmorata. AB - The electric organ of Torpedo marmorata contains a membrane-bound, captopril sensitive metallopeptidase that resembles mammalian angiotensin converting enzyme (peptidyl dipeptidase A; EC 3.4.15.1). The Torpedo enzyme has now been purified to apparent homogeneity from electric organ by a procedure involving affinity chromatography using the selective inhibitor lisinopril immobilised to Sepharose via a 28-A spacer arm. The purified protein, like the mammalian enzyme, acted as a peptidyl dipeptidase in cleaving dipeptides from the C-terminus of a variety of peptide substrates, including angiotensin I, bradykinin, [Met5]enkephalin, [Leu5]enkephalin, and the model substrate hippuryl (benzoylglycyl; BzGly)-His Leu. The hydrolysis of BzGly-His-Leu was activated by Cl-. Enzyme activity was inhibited by classical angiotensin converting enzyme inhibitors, including captopril, enalaprilat (MK422), and lisinopril (MK521). Torpedo angiotensin converting enzyme, like its mammalian counterpart, was also able to act as an endopeptidase in hydrolysing the amidated neuropeptide substance P. Hydrolysis of substance P occurred primarily at the Phe8-Gly9 bond with release of the C terminal tripeptide, Gly-Leu-MetNH2, and this hydrolysis was blocked by selective inhibitors. The Torpedo enzyme was recognised by a polyclonal antibody to pig kidney angiotensin converting enzyme on immunoelectrophoretic (Western) blot analysis. Thus, on the basis of substrate specificity, inhibitor sensitivity, and immunological criteria, the Torpedo enzyme closely resembles mammalian angiotensin converting enzyme. However, the Torpedo enzyme appears somewhat larger (Mr = 190,000) than the pig kidney enzyme (Mr = 180,000) on sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The endogenous peptide substrate(s) for Torpedo electric organ angiotensin converting enzyme and the physiological role of the enzyme in this tissue remain to be evaluated. PMID- 3027264 TI - Identification and characterization of a polypeptide from a lobster neurosecretory gland that induces cyclic GMP accumulation in lobster neuromuscular preparations. AB - Several observations suggest that cyclic GMP might regulate some aspect of neuromuscular physiology or metabolism in the lobster. Homarus americanus: lobster muscle is one of the richest known sources of cyclic GMP-dependent protein kinase, the preparation contains several phosphoproteins whose state of phosphorylation is affected by cyclic GMP more effectively than by cyclic AMP, and guanylate cyclase and phosphodiesterase are active in this tissue. However, no factor has yet been identified that alters lobster muscle cyclic GMP levels. We have screened extracts of neural and neurosecretory structures for the capacity to promote cyclic GMP accumulation in isolated exoskeletal muscles. Extracts of the sinus gland (a neurohemal organ found in the eyestalk) contain a factor that induces up to 100-fold increases in muscle cyclic GMP content, whereas extracts of other tissues are ineffective. This factor can also act on targets other than muscle, with hepatopancreas, testis, and neuronal tissue showing the largest responses. The sinus gland factor does not appear to affect cyclic GMP metabolism by depolarizing the preparation or by mobilizing extracellular Ca2+. The effect on cyclic GMP levels is dose-dependent and linear with time. Biological activity is destroyed by boiling and by 90% ethanol. It is also destroyed by trypsin, chymotrypsin, or pronase, which suggests that the factor is a protein or peptide. Both gel filtration chromatography and experiments using dialysis tubing with different molecular weight exclusion limits indicate that the factor has an apparent molecular weight of 5,000-12,000 daltons. A preliminary fractionation scheme, based on gel filtration, ion exchange, and reverse-phase chromatography, gives greater than 1,300-fold purification. Our long-range goal is to purify this factor to homogeneity, compare it to other peptide hormones, and use it as a probe to evaluate the role of cyclic GMP at the neuromuscular junction. PMID- 3027265 TI - Effects of chronic ethanol administration on rat brain phospholipid metabolism. AB - Alterations in brain phospholipid metabolism were observed after chronic ethanol administration for 16 days to developing rats. Animals were injected intraperitoneally with 32Pi 16 h prior to killing. Overall uptake of 32Pi by brain did not differ between the control and ethanol-treated groups, which were killed 2 h and 24 h after the last ethanol feeding. Except for an increase in the labeling of myelin after ethanol treatment, the amount of radioactivity recovered in the synaptosomal-mitochondrial and plasma membrane fractions of control and ethanol-treated groups was not different. Relative to the radioactivity of phosphatidylcholines, which indicated no change, there were increases (20-44%) in labeling of ethanolamine plasmalogens, phosphatidic acids, and phosphatidylinositols in cortical synaptosomes from the 2-h ethanol-treated group. In the plasma membrane fractions, however, increases (9-14%) in labeling of phosphatidylserines and phosphatidylinositols were observed in both 2- and 24 h ethanol-treated groups. In both membrane fractions, there was an obvious increase (44-86%) in labeling of polyphosphoinositides at 24 h after withdrawal from ethanol. Results thus indicate an adaptive increase in the biosynthesis of ethanolamine plasmalogen and brain acidic phospholipids due to chronic ethanol administration. Furthermore, the increase in labeling of polyphosphoinositides in the 24-h withdrawal group may reflect the hypoactivity associated with ethanol withdrawal. PMID- 3027266 TI - Cytochrome c oxidase deficiency in subacute necrotizing encephalomyelopathy. AB - Two new patients with Leigh's syndrome (subacute necrotizing encephalomyelopathy) due to deficiency of cytochrome c oxidase are presented and their data are compared with those of the four Leigh's syndrome patients previously reported with this deficiency. It is not possible to distinguish between the various biochemical aetiologies of Leigh's syndrome on clinical grounds. Investigation of pyruvate metabolism and of the respiratory chain will reveal the enzymatic defect in some of the patients. It has now been firmly established that a relationship exists between Leigh's syndrome and deficiency of cytochrome c oxidase. There are, however, other syndromes which are also associated with a deficiency of this enzyme. In Leigh's syndrome, the enzyme deficiency has been reported in many organ systems and in cultured fibroblasts. In the liver, however, decreased, intermediate or normal values of cytochrome c oxidase activity have been found. Selective or more widespread involvement of organ systems, due to mutations of either the nuclear or the mitochondrial DNA encoding for different subunits of the enzyme molecule (some of which may be organ- or tissue-specific), could explain the clinical and biochemical heterogeneity of syndromes associated with a cytochrome c oxidase deficiency. PMID- 3027267 TI - A randomized study of oral nutritional support versus ad lib nutritional intake during chemotherapy for advanced colorectal and non-small-cell lung cancer. AB - One hundred ninety-two patients with previously untreated metastatic cancer (102 non-small-cell lung cancer [NSCLC]; 90 colorectal cancer) were randomized to receive either ad lib nutritional intake (control group) or specific nutritional intervention during a 12-week study period when chemotherapy was administered. Those patients randomized to nutritional interventions were counselled to take oral nutrients with caloric intake equal to 1.7 to 1.95 times their basal energy expenditure, depending on their pretreatment nutritional status ("standard" group). An augmented group was counselled to have a caloric intake equivalent to that of the standard group but with 25% of calories provided as protein and additional supplements of zinc and magnesium. Counselling increased caloric intake in both tumor types but reduced weight loss in the short term only for lung cancer patients. Ninety-three NSCLC patients were evaluable for tumor response to vindesine and cisplatin. Overall, only 20.4% of the patients responded, and there were no significant differences in response rates, median time to progression, or overall duration of survival between the nutrition intervention groups and the control group. The tumor response rate to time sequenced 5-fluorouracil (5-FU) and methotrexate in the 81 evaluable patients with colorectal cancer was only 14.8%, and no significant differences in tumor response rates were noted between the three groups. Furthermore, the median time to progression and overall duration of survival were not different for the control, standard, and augmented groups. Nutritional interventions using dietary counselling had no impact on the percent of planned chemotherapy dose administered, the degree of toxicity experienced by patients, or the frequency of treatment delays. A multivariate prognostic factor analysis demonstrated that for lung cancer, the percent of weight loss, serum albumin concentration, and presence of liver metastases were significant (P less than .05) and independent prognostic variables for survival duration. For colorectal cancer, serum albumin, alkaline phosphatase, lactic dehydrogenase (LDH) levels, and percent targeted caloric intake (TCI) were significant independent predictors of survival duration. PMID- 3027268 TI - Carboplatin (Paraplatin; JM8) and etoposide (VP-16) as first-line combination therapy for small-cell lung cancer. AB - Fifty-two previously untreated patients with small-cell lung carcinoma (SCLC) were treated with a combination of carboplatin 300 mg/m2 intravenously (IV) on day 1 and etoposide 100 mg/m2 IV on days 1 through 3 every 28 days for four courses. Patients with limited disease (LD) subsequently received thoracic radiotherapy; no prophylactic cranial radiotherapy was used. Forty-four patients (85%) achieved an objective response, including 82% (29% complete remissions) of LD patients and 88% (13% complete remissions) of extensive-disease (ED) patients. Median response duration for LD patients was 7 months and 5.5 months for ED patients. Median survival for both LD and ED patients was 9.5 months. Myelosuppression was the main toxicity, with World Health Organization (WHO) grade 3/4 leucopenia occurring in 44% of patients. There was one (2%) treatment related neutropenic death. Treatment was otherwise well tolerated, and in particular no renal toxicity, neurotoxicity, or ototoxicity was seen. This new combination is highly active in terms of response rate, but response duration and survival is disappointing, and might be improved by prolonged treatment or by the use of additional drugs in combination. PMID- 3027269 TI - Metastatic patterns in small-cell lung cancer: correlation of autopsy findings with clinical parameters in 537 patients. AB - Postmortem material from 537 patients included in various protocols of intensive combination chemotherapy or chemoradiotherapy for the management of small-cell anaplastic carcinoma of the lung (SCLC) has been reviewed. Patterns of residual or recurrent disease were analyzed in relation to pretreatment clinical parameters. Residual primary tumor (P less than .05), regional lymph node involvement (P less than .01), hepatic (P less than .01), bone (P less than .05), and renal metastases (P less than .05) were all significantly less frequent among patients with initially limited-stage disease compared with extensively staged patients. The frequency of residual intrathoracic tumor and metastatic pattern did not significantly differ between partial responders (PRs) and nonresponders (NRs). Patients with limited disease achieving a complete remission had a lower frequency of intrathoracic tumor (P less than .001) at autopsy compared with limited-stage PRs. However, for extensive-stage patients the pattern of residual disease was essentially independent of tumor response. Prior surgery was associated with a reduced burden of metastases only in those who underwent a radical resection. The addition of radiotherapy to the primary tumor and mediastinum failed to modify the autopsy distribution of residual tumor compared with that in patients treated with chemotherapy alone. Metastatic patterns were similar with or without prophylactic abdominal radiotherapy, while prophylactic cranial irradiation (PCl) did not prevent the development of cerebral metastases in patients whose systemic response to treatment was either partial or nonexistent. However, a beneficial effect of PCl in compete responders (CRs) was not excluded. PMID- 3027270 TI - Detection of liver metastases in small-cell lung cancer: a comparison of peritoneoscopy with liver biopsy and ultrasonography with fine-needle aspiration. AB - Liver evaluation of 131 patients with small-cell lung cancer (SCLC) was performed both by peritoneoscopy (PS) with liver biopsy and by ultrasonography (US) with fine-needle aspiration. A total of 33 patients (25%) had liver involvement, 82% detected by US and 76% detected by PS. The difference was due to 27 incomplete investigations by PS and two incomplete investigations by US. In 104 patients in whom both investigations were "successful," PS confirmed 86% and US confirmed 79% of the patients with liver metastases. In each of the investigations, 7% (PS) and 14% (US) of patients had false-negative conclusions as compared with histologic evidence obtained by the other method. US found six patients with extrahepatic intraabdominal disease, while PS found none. S-lactic dehydrogenase (s-LDH), SGOT, and s-alkaline phosphatase were found to be too unspecific to indicate liver metastases unless all three tests were normal or abnormal. It is recommended that US should be used as the initial procedure when staging patients with SCLC, and that PS can be considered complementary in patients with negative US. PMID- 3027271 TI - A phase I-II study of bialkylator chemotherapy, high-dose thiotepa, and cyclophosphamide with autologous bone marrow reinfusion in patients with advanced cancer. AB - Twenty patients with disseminated cancer both untreated and previously treated received bialkylator chemotherapy, thiotepa, and cyclophosphamide and reinfusion of cryopreserved autologous bone marrow (ABMR). The cyclophosphamide dose was constant at 7.5 g/m2 over three days, while thiotepa was started at 1.8 mg/kg for three days in escalating dose by a modified Fibonacci schema to 7 mg/kg. The median time to recovery of more than 500 granulocytes and more than 50,000 platelets/microL was 18 and 27 days, respectively. Four patients died as a consequence of severe, overwhelming infections or progressive disease during their period of aplasia. Of the 18 evaluable patients, a complete response (CR) was achieved in three patients and a partial response (PR) in ten patients for an overall response rate of 72%. The median duration of response was 14 weeks. Other nonhematologic toxicities included nausea/vomiting, diarrhea, stomatitis, skin rash, and cardiomyopathy. The maximum tolerated dose (MTD) of thiotepa was 700 mg/m2 or 6 mg/kg for three doses. Although there are substantial toxicities associated with this regimen, high-dose thiotepa and cyclophosphamide produce high response rates in patients with disseminated cancer. PMID- 3027273 TI - Ventroposterior thalamic regions projecting to cytoarchitectonic areas 3a and 3b in the cat. AB - The adequate stimulus and body site that excited neurons in cat cortical somatosensory areas 3a and 3b were recorded using low-impedance tungsten microelectrodes. Horizontal penetrations provided a good correlation between the electrophysiological and cytoarchitectonic data. Responses best driven by cutaneous stimuli were replaced with responses driven by manipulation of deep tissue at, or very near, the border between areas 3a and 3b. Following functional identification of these areas horseradish peroxidase was injected into one of them. Injections into area 3a labeled neurons in a rostral and dorsal cap of the ventroposterior thalamus. It was suggested that this region is a distinct nucleus termed the ventroposterior oralis nucleus (VPO). Injections into the forelimb portion of area 3b labeled neurons in the ventroposterior lateral nucleus (VPL). With both vertical and horizontal microelectrode trajectories through the ventroposterior thalamic nuclei, inputs from deep structures presumed to be muscles were consistently located in the VPO nucleus and cutaneous inputs activated neurons in the VPL. The existence of several functionally-distinct subdivisions within the somatosensory nuclei of the thalamus supports the hypothesis of parallel processing and relay of somatosensory information at this level of the pathway. PMID- 3027272 TI - A thalamic terminus of the lateral cervical nucleus: the lateral division of the posterior nuclear group. AB - The submodality and receptive field properties of single units in the lateral cervical nucleus (LCN) of barbiturate anesthetized cats were studied with glass microelectrodes. In other experiments, a region of the posterior thalamus containing neurons with properties comparable to those seen in the LCN was examined with tungsten microelectrodes. The responses of most units in the LCN reflected a major input from large myelinated afferent fibers innervating guard hairs but no input from Pacinian afferent fibers. The large size of the receptive fields indicated that excitatory input converged selectively from afferent fibers serving hairs over large areas of the body. In the posterior thalamus rapidly adapting neurons characterized by very large receptive fields and driven by the movement of guard hairs were observed to a region identified histologically as the rostral extension of the lateral division of the posterior nuclear group (POl). Caudally this region was located immediately adjacent to the dorsolateral part of the ventroposterior inferior nucleus (VPI). In more rostral parts of the thalamus it was located more dorsally and the ventroposterior lateral nucleus intervened between it and the VPI. This region was less than 1 mm wide in the frontal plane but extended rostrocaudally for several millimeters. Horseradish peroxidase injected into the region of the VPI and the POl labeled many cells in the LCN and the caudal pole of the dorsal column nuclei demonstrating that neurons in the LCN relay information to this part of the thalamus. These data, plus previous experiments showing that the VPI receives a major projection from the caudal poles of the dorsal column nuclei, suggest that the rostral portion of the POl receives an important afferent supply from the LCN. The responses of neurons in the POl appear to arise from specific classes of sensory receptors and cannot be considered less precise or more primitive than responses observed in the ventroposterior nucleus of the thalamus. PMID- 3027275 TI - Distribution of cAMP and cAMP-dependent protein kinases in Aplysia sensory neurons. AB - Sensitization of the gill- and siphon-withdrawal reflex in Aplysia is considered a simple form of learning. Previous work has provided physiological and pharmacological evidence that cAMP-dependent protein phosphorylation within identified sensory neurons of the abdominal ganglion underlies the short-term form of this behavioral modification. Our main goal in this paper is to determine the subcellular distribution of cAMP and to measure the amounts and properties of the 2 types of subunits (regulatory and catalytic) that constitute the cAMP dependent protein kinase. Do these biochemical parameters differ in sensory cells from those in other parts of nervous tissue? We found that the increased cAMP synthesized under conditions of sensitization is distributed in 3 compartments in the neuron: most of it is free in the cytoplasm; the remainder is bound either to cytoplasmic or to particulate proteins, which are believed to be regulatory subunits of the cAMP-dependent protein kinase. Binding of cAMP within the neurons is a measure of activation of the kinase. At rest, 17% of the binding sites in sensory cells were occupied. After brief electrical stimulation of the connective, which released endogenous transmitter, occupancy increased to 34%. This treatment increased the amount of cAMP bound to the various binding proteins differentially. The biochemical characteristics of cAMP binding were found to be the same in sensory neurons as in the rest of the nervous system but different from those in muscle. Thus, memory and learning are likely to be mediated by enzymes that are shared by other nerve cells. We found that sensory neurons have greater cAMP-dependent protein kinase activity than other neurons, however, and as a result may be more sensitive to small increases of cAMP. PMID- 3027274 TI - The dendritic origins of penicillin-induced epileptogenesis in CA3 hippocampal pyramidal cells. AB - Experiments were performed in order to identify the sites of epileptiform burst generation in rat hippocampal CA3 pyramidal cells. A subsequent slow field potential was studied, which is associated with afterdischarge generation. Laminar field potential and current source-density (CSD) methods were employed in hippocampal slices exposed to penicillin. Simultaneous intracellular and extracellular field recordings from the CA3 pyramidal cell body layer showed that whenever an epileptiform burst was recorded extracellularly, individual CA3 neurons underwent an intense depolarization shift. In extracellular records a slow negative field potential invariably followed epileptiform burst generation. In approximately 10% of slices, synchronous afterdischarges rode on the envelope of this negative field potential. Intracellularly a depolarizing afterpotential followed the depolarization shift and was coincident with the extracellular slow negative field potential. A one-dimensional CSD analysis performed perpendicular to the CA3 cell body layer showed that during epileptiform burst generation large current sinks occur simultaneously in the central portions of both the apical and basilar dendrites. The average distance of the peak amplitude for these sinks from the center of the cell body layer was 175 +/- 46.8 microns and 158 +/- 25.0 microns, respectively. A large current source was recorded in the cell body layer. Smaller current sources were observed in the distal portions of the dendritic layers. During the postburst slow field potential a current sink was recorded at the edge of the cell body layer in stratum oriens--a region referred to as the infrapyramidal zone. Simultaneous with the current sink recorded there, smaller sinks were often observed in the dendritic layers that appeared to be "tails" or prolongations of the currents underlying burst generation. Two dimensional analyses of these field potentials were performed on planes parallel and perpendicular to the exposed surface of the slice. Isopotential contours showed that the direction of extracellular current is mainly orthogonal to the CA3 laminae. Correction of CSD estimates made perpendicular to the cell body layer for current flowing in the other direction did not alter the location of computed current sources and sinks. In order to show that the dendritic currents associated with epileptiform burst generation were active sinks, tetrodotoxin (TTX) was applied locally to the dendrites where the current sinks were recorded.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3027276 TI - Ontogeny, compartmentation, and turnover of spectrin isoforms in rat central neurons. AB - A variant of a principal structural protein of erythrocytes, spectrin, is a major neuronal protein. Here we have examined the subcellular and regional distributions, subunit composition, ontogeny, and metabolism of spectrin in rat CNS. While all subcellular fractions, except the mitochondrial, expressed the previously characterized brain form of spectrin (fodrin, or alpha gamma spectrin), limited brain regions contained, in cytoplasm, a second isoform immunologically related to erythrocyte alpha beta-spectrin. Both alpha gamma- and alpha beta-spectrin are primarily neuronal, as evidenced by immunocytochemistry. The spectrins are distributed between 2 distinct subneuronal compartments: a membrane-associated domain containing alpha gamma-spectrin in relatively constant amounts across brain regions, and a cytoplasmic domain containing both the alpha gamma and alpha beta isoforms in widely varying amounts across brain regions. Although forebrain has considerable alpha beta-spectrin, the diencephalon, mesencephalon, and brain stem are devoid of this isoform. Further evidence for spectrin compartmentation comes from its ontogeny. Membrane-associated alpha gamma-spectrin is present at birth at its adult levels, but cytoplasmic alpha beta-spectrin is expressed only following the second postnatal week. Similarly, the 4-fold difference in cytoplasmic alpha gamma-spectrin content across brain regions develops during the third postnatal week. In this compartment, both spectrin forms may be metabolized in vivo, at least in part, by calcium-activated proteolysis. The presence in mammalian neurons of 2 spectrin isoforms and their compartmentation into distinct domains suggests multiple functions for neuronal spectrin, one of which may be in the stabilization or maturation of forebrain neurons. PMID- 3027277 TI - Coexistence of GABA receptors and GABA-modulin in primary cultures of rat cerebellar granule cells. AB - GABA-modulin (GM), a basic polypeptide purified from rat brain synaptosomes, which is an allosteric inhibitor of GABA recognition sites, has been detected in primary cultures of cerebellar interneurons enriched in granule cells by immunohistochemistry, using a specific antibody raised in rabbit injected with GM purified from rat brain synaptosomes. In these cultures, GM is expressed by the granule cells, which are postsynaptic to GABAergic interneurons, but not by glial cells. In rat cerebellar sections anti-GM antiserum intensely strains the granular cell layer and Purkinje cell dendrites and cell bodies. GM has been purified from the cerebellar granule cell cultures and appears to be identical under biochemical, immunological, and functional criteria to authentic GM purified from rat brain synaptosomes. Granule cell cultures devoid of GABAergic neurons contain the GABA/BZ/Cl- receptor complex; in fact, intact cell monolayers, incubated in physiological buffer at 25 degrees C, express 3H muscimol and 3H-flunitrazepam binding sites, which are comparable to the sites detected in cell membrane preparations and which modulate each other reciprocally. It is concluded that GM might participate in the supramolecular organization of the GABA receptor complex, perhaps functioning as a modulator of this receptor protein. PMID- 3027278 TI - A model of chronic pain in the rat: functional correlates of alterations in the activity of opioid systems. AB - Intradermal inoculation of rats at the tail base with Mycobacterium butyricum led to the gradual development of an arthritic swelling of the limbs which peaked at 3 weeks and subsided thereafter. Arthritic rats displayed a loss of body weight, hypophagia, and hypodipsia in addition to a disruption of the diurnal rhythms of ingestive behavior and of core temperature. The activity of adenohypophyseal beta endorphin-(beta-EP) secreting corticotrophs, in contrast to prolactin-(PRL) secreting lactotrophs, was increased in arthritic rats. Indeed, hypertrophy of the adrenal glands was seen. Arthritic rats also showed an elevation in spinal cord levels of immunoreactive dynorphin (DYN), an endogenous ligand of the kappa opioid receptor. The paws and tail of arthritic rats showed lower thresholds in response to noxious pressure (hyperalgesia), higher thresholds in response to noxious heat (hypoalgesia), and no change in their response to noxious electrical stimulation. Neither naloxone nor ICI-154, 129 (a preferential delta-receptor antagonist) modified the responses of the paw or tail to pressure. However, MR 2266 (an antagonist with higher activity at kappa-receptors) decreased thresholds to pressure in arthritic, but not control, rats; that is, it potentiated the hyperalgesia. This action was stereospecific. None of the antagonists modified the response to heat. MR 2266 did not affect the response to pressure in rats with acute inflammation produced by yeast. Thus, the potentiation of pressure hyperalgesia by MR 2266 in chronic arthritic rats is highly selective. Arthritic rats showed a reduced response to the analgesic effect of a kappa-agonist (U 50,488H), whereas the response to a mu-agonist (morphine) was enhanced. These effects were specific to nociception in that their influence upon endocrine secretion (PRL and beta-EP) was otherwise changed. The secretion of beta-EP and PRL was stimulated by both morphine and U-50,488H, and the influence of U-50,488H upon the release of beta-EP (from the adenohypophysis) was enhanced in arthritic rats. It is suggested that polyarthritis is a complex condition entailing many changes, both behavioral and endocrinological. Further, arthritic rats cannot simply be described as "hyperalgesic": of critical importance is the nature of the nociceptive stimulus applied. The parallel alterations in spinal cord pools of DYN and kappa-receptors (see also Millan et al., 1986) and the changes in the influence on nociception of kappa-agonists and kappa-antagonists suggest an increased activity of spinal DYN. Thus, spinal kappa-receptors may play a role in the modulation of nociception under chronic pain.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3027279 TI - Corticotropin-releasing factor receptors in the rat central nervous system: characterization and regional distribution. AB - A stable, iodine-125-labeled analog of rat/human corticotropin-releasing factor (CRF) was used to define the characteristics of CRF receptors in a crude mitochondrial/synaptosomal membrane preparation of rat olfactory bulb, and to study the distribution of CRF binding sites in discrete regions of the rat CNS. The binding of 125I-Tyro rat/human CRF (125I-rCRF) was time- and temperature dependent, was sensitive to the pH, ionic strength, and cationic composition of the incubation buffer, and was linear over a broad range of membrane protein concentrations. 125I-rCRF binding to olfactory bulb membrane was saturable, reversible, and, on Scatchard analysis, revealed a high-affinity component with an apparent equilibrium dissociation constant (Kd) of 0.2 nM and a low-affinity binding site with Kd of approximately 20 nM. Data from pharmacological studies indicated that the ability of a variety of CRF fragments and analogs to inhibit 125I-rCRF to olfactory bulb membranes correlates well with their reported relative potencies in stimulating pituitary adrenocorticotropic hormone secretion in vitro. Consistent with a coupling of CRF receptors to adenylate cyclase, the binding of 125I-rCRF was decreased by guanine nucleotides and increased by magnesium ions. A heterogeneous distribution of 125I-rCRF binding sites was found in the rat CNS, with highest densities present in olfactory bulb, cerebellum, cerebral cortex and striatum, and progressively lower but significant levels of binding were detected in cervical spinal cord, hypothalamus, medulla, midbrain, thalamus, pons, and hippocampus. These data, using a rat CRF ligand homologous to the endogenous peptide, are consistent with those from previous studies demonstrating the presence of specific binding sites for ovine CRF in rat brain, and provide further support for the suggestion that endogenous CRF may function as a neurotransmitter in the CNS. PMID- 3027281 TI - Technetium-99m(V)-DMSA in the imaging of medullary thyroid carcinoma. PMID- 3027280 TI - Cerebral blood flow imaging with thallium-201 diethyldithiocarbamate SPECT. AB - Thallium-201 diethyldithiocarbamate ([201TI]DDC) was studied in humans as an agent for cerebral blood flow imaging. Brain uptake proved to be complete 90 sec after injection with no appreciable washout or redistribution for hours. Intracarotid injection suggested an almost 100% extraction during the first passage. Whole-body distribution studies demonstrated a brain uptake of 4.3% of the dose compared with 0.9% for [201TI]chloride. No differences were found in the distribution of [201TI]DDC versus [201TI]chloride in other organs. After the injection of 3 mCi 201TI, good quality single photon emission computed tomographic (SPECT) images of the brain were obtained with both a rotating gamma camera and a multidetector system. In ischemic brain disease, perfusion defects were easily demonstrated. We conclude that [201TI]DDC is a suitable radiopharmaceutical for SPECT studies of cerebral blood flow. PMID- 3027282 TI - Effects of purified dietary fiber sources on beta-carotene utilization by the chick. AB - Effects of various purified dietary fiber components on beta-carotene utilization by the chick were investigated in two experiments (expt.). Eight-day-old Columbian X New Hampshire male (expt. 1) or female (expt. 2) chicks were fed a vitamin A-deficient diet for 1 wk and then fed beta-carotene-supplemented diets containing 0% fiber, 7% arenaceous flour or 7% of a purified fiber source for 4 wk. Results of expt. 1 showed that hemicellulose, lignin and citrus pectin, but not arenaceous flour or polygalacturonic acid, depressed beta-carotene utilization by the chick, as measured by percentage of consumed beta-carotene stored in liver as vitamin A relative to the 0% fiber control. In expt. 2, effects of the methoxyl content of pectin were studied. High and medium methoxyl apple pectin, citrus pectin and polygalacturonic acid reduced storage of vitamin A in liver. Low methoxyl apple pectin had no significant effect on beta-carotene utilization. Thus, several purified forms of dietary fiber significantly reduced beta-carotene utilization by chicks when fed at the 7% supplementary level. Moreover, with pectin, there was an inverse relationship between methoxyl content of pectin and beta-carotene utilization. PMID- 3027283 TI - Effects of indigestible dietary bulk and short chain fatty acids on the tissue weight and epithelial cell proliferation rate of the digestive tract in rats. AB - Influences of indigestible dietary bulk (kaolin), short chain fatty acids (SCFA), and their interaction on the weight and epithelial cell proliferation rate of digestive organs were studied. Rats with an ileal fistua were fed an elemental diet with or without kaolin (10% w/w). They were given a 3 ml SCFA mixture (acetate, propionate, and butyrate; 100, 20, and 60 mM, respectively; pH 6.1) or a 3 ml NaCl solution (180 mM; pH 6.1) via the fistua twice a day for 14 days. Kaolin, but not SCFA, increased full and empty body weight by 14% and 13%, respectively. Kaolin increased the tissue weight, but not relative tissue weight (tissue weight/empty body weight), of the forestomach, distal colon, liver and heart by 6-14%. SCFA, independent of the presence of kaolin, increased the weight and the relative weight of the cecum, both by 15%. SCFA, but not kaolin, increased the cryptal cell production rate in colonic segments by 50-140%. These results indicate that SCFA accelerates epithelial cell proliferation, thereby increasing cecal tissue weight, kaolin stimulates body weight gain which is accompanied by a proportional increase in the tissue weight of some digestive organs, and the actions of these factors are independent of each other. PMID- 3027284 TI - New screening device for assessment of peripheral neuropathy. AB - Screening for the onset of toxic and entrapment neuropathy is a major concern in occupational medicine today. Current perception thresholds may be used as a measure of the integrity of the peripheral nervous system. A new transcutaneous nerve stimulator was used to evaluate current perception thresholds in 54 normal persons and 33 diabetic subjects. Current perception thresholds in the normal volunteers with no evidence of peripheral neuropathy were found to vary significantly with the frequency and location of the stimulation, as well as age. The test identified the diabetic peripheral neuropathy with an overall sensitivity of 94%. This new diagnostic technique is quick, simple to perform, noninvasive and nonaversive and provides a sensitive quantitative measure of sensory function. This diagnostic stimulator will be useful for screening occupationally related toxic and entrapment neuropathies in which sensory impairment is an early finding. PMID- 3027285 TI - Osteogenesis in replamineform hydroxylapatite porous (RHAP) ceramic implants used for human mandibular ridge augmentation: report of two cases. PMID- 3027286 TI - Effect of proteolytic enzymes on caries lesion formation in vitro. AB - This study investigates enzyme degradation of collagen at various pH levels and is designed to obtain information pertaining to the effect of proteolytic enzymes on caries-like lesion formation and progression. One hundred and eight sections of human teeth with exposed areas on enamel and root face, were placed into an enzyme/buffer system maintained at pH levels of 4.0, 5.5 and 7.0. Periodically, throughout the experimental period, sections were removed and examined histologically for caries-like lesion formation. Results showed consistent surface erosion occurring on the root face only at a pH level of 5.5 in buffer containing enzymes. Sections placed in pH 5.5 buffer only, which served as controls, showed obvious lesion formation on the root face, but with histologically intact surfaces. This interaction occurring between enzymatic activity and acid demineralization suggests that enzymes may have a contributory effect on caries lesion formation in root surfaces. PMID- 3027287 TI - Pigmentary changes in rat oral mucosa following antimalarial therapy. AB - Pharmacogenic pigmentation of the oral mucosa has been reported following the use of a number of anti-malarial drugs. The nature and distribution of the pigment is inconclusive in the literature. The aim of the present study was to document pigment deposition within the oral mucosa of DA rats following prolonged chloroquine and pyrimethamine administration. The drugs were given as a combined dosage and separately to different groups via stomach gavage tube. After 12 weekly administrations the palatal mucosa was examined histochemically and ultrastructurally for changes in numbers and size of active melanocytes using the dopa-oxidase technique. The serum was analysed for changes in ACTH and testosterone levels. Morphometric analysis of cells incubated for dopa-oxidase showed a significant increase in the size of dopa positive cells with both drugs but an increase in the number of active melanocytes with chloroquine only. Serum levels of ACTH remained unchanged with both drugs but pyrimethamine caused an elevation in testosterone. PMID- 3027288 TI - Human papillomavirus type 2 DNA in oral verrucous carcinoma. AB - Tissues from patients with oral verrucous carcinoma were examined for the presence of human papillomavirus (HPV). The tissues were stained for the presence of the type common papillomavirus antigen by immunohistochemical staining and the presence of HPV DNA was determined by in situ hybridization with biotin-labelled HPV DNA probes. Seventeen tissue specimens were obtained from 9 patients, and included pre-malignant lesions and primary and recurrent tumors. One pre malignant lesion was positive for papillomavirus structural antigen. This lesion and lesions from 2 other patients hybridized at low stringency (Tm-35 degrees) to 3 different HPV probes. By hybridization under high stringency conditions (Tm-20 degrees), the virus in each case was identified as being HPV2. PMID- 3027289 TI - Endocrine differentiation of extra-pulmonary small cell carcinoma demonstrated by immunohistochemistry using antibodies to PGP 9.5, neuron-specific enolase and the C-flanking peptide of human pro-bombesin. AB - Several recent studies have confirmed the endocrine nature of small cell carcinoma of the lung. In extra-pulmonary sites, small cell 'undifferentiated' carcinomas have classical morphological features similar to their pulmonary counterpart. We therefore investigated, using immunocytochemistry, the possibility that the non-pulmonary neoplasms may also be endocrine in nature. Sections of 29 small cell carcinomas from oesophagus, stomach, larynx, colon and urinary bladder were immunostained using antisera to protein gene product 9.5 (PGP 9.5), neuron-specific enolase (NSE), cytokeratin, leucocyte common antigen and peptides including bombesin, the C-flanking peptide of human probombesin, adrenocorticotrophic hormone, neurotensin, calcitonin and pancreatic polypeptide. All the tumours showed immunoreactivity for at least one of the two general endocrine markers PGP 9.5 and NSE. Twenty-three of the 29 cases were immunoreactive for PGP 9.5, 27 for NSE. All were positive for cytokeratin and negative for leucocyte common antigen. Of the regulatory peptides, immunoreactivity was obtained with antisera to bombesin (one case), the C flanking peptide of human pro-bombesin (14 cases), adrenocorticotrophic hormone (one case) and calcitonin (three cases). No PGP 9.5-, NSE- or peptide-like immunoreactivity was detected in 25 control tumours from similar sites, including lymphomas and poorly differentiated tumours. These results suggest that non pulmonary small cell carcinoma has an endocrine character. PMID- 3027290 TI - Immunocytochemical demonstration and significance of p21 ras family oncogene product in benign and malignant breast disease. AB - It has been suggested that the immunocytochemical demonstration of the p21 ras oncogene product is a useful marker of malignancy in breast disease. We have studied the reactivity of a series of specimens of benign and malignant breast disease with the anti ras p21 monoclonal antibody Y13-259, and shown widespread positive staining in both benign and malignant (including metastatic) disease as well as in adjacent 'normal' epithelium. In addition some staining of stromal cells as well as nerve fibres was observed. Our results suggest that the presence of ras p21 protein as demonstrated by this antibody is not a useful marker of malignancy or of proliferating epithelium but is rather a normal feature of certain cell types. PMID- 3027291 TI - Breast carcinoma with stromal multinucleated giant cells--a light microscopic, histochemical and ultrastructural study. AB - A rare form of infiltrating ductal carcinoma of the breast containing numerous benign stromal multinucleated giant cells (MNGC) is described. Giant cell tumours of the breast are usually the result of stromal metaplasia or fusion of malignant cells or occur as extraskeletal giant cell tumours. Benign multinucleated cells in breast carcinoma, however, are a very unusual phenomenon and have been said to arise from the fusion of mononuclear cells, in response to increased vascularity. The present investigation by light and electron microscopy, in part, supports an origin for the multinucleated giant cells from mononuclear cells, but immunohistochemistry surprisingly failed to confirm this observation. Also, the formation of the multinucleated giant cells did not show any direct relationship with tumour vascularity. PMID- 3027292 TI - Nuclear DNA content in breast carcinomas with neuroendocrine differentiation. AB - Sixty-one breast carcinomas (54 infiltrating ductal carcinomas and seven infiltrating lobular carcinomas) were immunostained with anti-NSE and analysed with respect to nuclear DNA content. Nine of the 23 NSE-positive breast carcinomas were diploid, five were triploid, six tetraploid and three pentaploid. Twenty-one of the 38 NSE-negative tumours were diploid, 10 were triploid, seven tetraploid, and none were pentaploid. Three of the eight histologically grade I tumours in the NSE-positive group were aneuploid, whereas all the six grade I tumours in the NSE-negative group were diploid. The results show that a proportion of breast carcinomas with neuroendocrine differentiation are aneuploid and that aneuploid tumours that are grade I histologically are found in the NSE positive group and not in the NSE-negative group. PMID- 3027294 TI - Restriction endonuclease analysis of repetitive sequences in the Trichinella genome: three strain-specific patterns. PMID- 3027293 TI - Clinical presentation of mitochondrial respiratory chain defects in NADH-coenzyme Q reductase and cytochrome oxidase: clues to pathogenesis of Leigh disease. AB - Measurement of pyruvate and lactate produced from glucose by confluent skin fibroblast cultures from 95 patients with lactic acidemia revealed 10 in whom the lactate/pyruvate ratio (L/P) was increased (L/P = 57 to 232) compared with that observed in control cell lines (L/P = 18 to 35). Mitochondria prepared from these cells revealed two types of respiratory chain defect. In four patients the deficient activity was present in NADH-coenzyme Q reductase (14% to 21% of controls), and in six the deficiency was in cytochrome c oxidase (21% to 28% of controls). The four patients with NADH-coQ reductase deficiency presented early with lactic acidosis, respiratory failure, anorexia, and hypotonia; all four died within 7 months. The group with cytochrome oxidase deficiency had a somewhat later (18 months to 2 years of age) presentation with milder lactic acidemia, but also with hypotonia and anorexia. They had delayed development, beginning to walk and talk at 18 to 24 months, and then slowly regressed. Although an investigation of central nervous system disorders in this latter group has not been possible, the clinical progression fits into the broad category of Leigh disease. We conclude that in these two groups respiratory chain defects can be detected and localized by the use of skin fibroblast cultures. PMID- 3027295 TI - Alkaline phosphatase and phosphodiesterase activities of the brush border membrane of four strains of the tapeworm, Hymenolepis diminuta. PMID- 3027297 TI - Self-resistance of the nourseothricin-producing strain Streptomyces noursei. AB - The nourseothricin producer Streptomyces noursei is resistant to its own antibiotic in submerged as well as in surface culture. The strain shows no cross resistance to miscoding inducing aminoglycoside antibiotics. Cell free extracts of Streptomyces noursei inactivate nourseothricin by enzymatic acetylation. The pattern of cross-resistance of Streptomyces noursei correlates well with the substrate specificity of the nourseothricin acetyltransferase. Furthermore, the acetyltransferase activity parallels the resistance level in nourseothricin producing strains and nonproducing mutants. The results suggest that the nourseothricin acetyltransferase is important in the self-defence strategy of the nourseothricin-producing strain. PMID- 3027296 TI - Infantile agranulocytosis with survival into adolescence: periodontal manifestations and laboratory findings. A case report. AB - A case of infantile agranulocytosis with survival into adolescence is presented. The polymorphonuclear leukocyte is considered an important source of lysosomal enzymes in gingival crevicular fluid, and evaluation of connective tissue degrading enzymes in the fluid was performed. The activity of beta-glucuronidase, a ground substance-degrading enzyme that may serve as a marker for polymorphonuclear leukocytes, was markedly reduced in the fluid compared to samples from systemically healthy adults with periodontitis. The activities of the ground substance-degrading enzyme arylsulfatase, and collagenase, were in the low-normal range. The plaque microbiology, as characterized by dark-field microscopy and selective culturing, was consistent with advanced periodontitis. A review of the medical history revealed a series of bacterial infections since infancy. Improvement in the systemic health of the patient occurred at about the age of 15, and the intake of antibiotics to control infections was correspondingly reduced after this time. An exacerbation of the patient's periodontal disease, as evaluated by loss of alveolar bone on radiographs, occurred 1 to 2 years later. The progression of periodontal disease observed in this patient was apparently associated with the withdrawal of antibiotics administered for control of systemic (nonoral) infections. PMID- 3027298 TI - [Preparation of the anterior edentulous ridge during extractions]. PMID- 3027299 TI - Relationship between phosphatidylinositol turnover and Ca++ mobilization induced by alpha-1 adrenoceptor stimulation in the rat aorta. AB - In this study the causal relationship between alpha-1 adrenoceptor activation mediating contraction in rat aorta and the mediatory responses, such as phosphatidylinositol turnover and intracellular Ca++ release has been evaluated. Norepinephrine (1 X 10(-5) M) increased maximally the accumulation of [3H]inositol-1-PO4. In the presence of LiCl (10 mM) the norepinephrine-induced accumulation of [3H]inositol-1-PO4 occurred in a time-dependent, linear fashion (0-60 min), achieving a 13-fold increase over the unstimulated control at 60 min of exposure. This stimulation could be inhibited by prazosin (1 X 10(-7) M) but not by yohimbine (1 X 10(-7) M), whereas it was also unaffected by nifedipine (3 X 10(-7) M). Potassium depolarization did not invoke [3H]inositol-1-PO4 production nor did Sgd 101/75 in concentrations of up to 3 X 10(-5) M, although both have been found effective in stimulating a large influx of Ca++ for their contraction. However, the effect of norepinephrine on the formation of [3H] inositol-1-PO4 was antagonized by Sgd 101/75. A positive correlation (correlation coefficient 0.966) between intracellular Ca++ release and phosphatidylinositol turnover induced by a series of alpha-1 adrenoceptor agonists was demonstrated. These data support the hypothesis that stimulation of alpha-1 adrenoceptors in rat aorta can elicit two distinct processes of Ca++ utilization for contraction. One facilitates exclusively an influx of extracellular Ca++ which is independent of phosphatidylinositol turnover, whereas the other activates the release of intracellularly bound Ca++ that may be mediated primarily by phosphatidylinositol metabolism. PMID- 3027300 TI - Diuretic actions in man of a selective kappa opioid agonist: U-62,066E. AB - The effect of a selective kappa opioid agonist, U-62,066E, on urine formation in human volunteers was assessed. Volunteers received single intramuscular injections of either placebo or 2, 3, 4, 5 or 6 micrograms/kg of U-62,066E in a randomized, double-blind study. U-62,066E caused dose-dependent maximal increases in urine volume of 2.6 times control in the first 4 hr after administration. A corresponding decrease in urine osmolality to 20% of base-line values occurred. No changes in total urinary Na, K or Cl excretion were identified. Kappa agonists produce a water-only diuresis at low doses in humans. The mechanism of this effect was not examined in this study but probably relates to alterations in antidiuretic hormone activity. PMID- 3027301 TI - Naloxazone treatment in the guinea pig ileum in vitro reveals second functional opioid receptor site. AB - Guinea pig ilea were incubated in vitro with naloxazone, an irreversible opioid receptor antagonist, in an attempt to determine a dissociation constant (KA) value for the opioid agonist, BW 942C, by the method of partial receptor alkylation. However, concentrations of naloxazone up to 3 X 10(-4) M (120 min incubation-60 min washout) did not depress the maximal response or the slope of the concentration-response curve for BW 942C and, therefore, did not allow calculation of KA value. To analyze the residual activity of BW 942C, tissues were incubated with naloxone for 60 min after naloxazone treatment. Schild analysis of naloxone antagonism after 3 X 10(-6) M naloxazone produced a pKB of 8.0 +/- 0.06 and a slope of 0.88 +/- 0.03, suggesting simple competitive antagonism at a single site. In the absence of naloxazone treatment, naloxone 10( 8) to 10(-5) M antagonized the effects of BW 942C yielding a pKB value and slope of 8.34 +/- 0.08 and 1.0 +/- 0.04, respectively. Schild analysis of naloxone antagonism after 10(-4) M naloxazone yielded a pA2 of 6.23 +/- 0.20 and a slope of 0.55 +/- 0.08. These values were not consistent with simple competitive antagonism at a single receptor site. Modeled curves showed that the data for naloxone antagonism after 10(-4) M naloxazone were consistent with action at two receptor sites with naloxone pKB values of approximately 8.3 (experimentally determined) and approximately 6.0. An alternative explanation of altered affinity for BW 942C and naloxone at a single site produced by naloxazone treatment cannot be excluded. PMID- 3027302 TI - Continuous intrathecal opioid analgesia: tolerance and cross-tolerance of mu and delta spinal opioid receptors. AB - The tolerance effects of continuous intrathecal infusions of opioids at mu and delta receptors were studied in rats. These effects were compared to those of chronic systemic morphine. A chronic intrathecal infusion of the relatively selective delta agonist, [D-Ala2, D-Leu5]enkephalin (DADLE), produced a larger degree of tolerance to DADLE than to the highly specific mu-activating morphiceptin analog [N-methyl-Phe3, D-Pro4]morphiceptin (PL017). The slope of the analgesic dose-response curve for the highly specific delta agonist, cyclic [D Penicillamine2, D-Penicillamine5]enkephalin (DPDPE), was significantly different (flatter) from those of mu agonists or DADLE. High-dose infusion of PL017 induced a 61-fold parallel shift of the dose-response curve for PL017. This same treatment also induced a corresponding flattened, nonparallel change of the dose response curve for DADLE. This altered curve for DADLE was very similar in slope to that of DPDPE. Pretreatment with the irreversible mu antagonist, beta funaltrexamine, caused a parallel rightward shift of the dose-response curve for PL017 but did not affect DPDPE activity. beta-Funaltrexamine treatment induced a nonparallel rightward shift of the dose-response curve for DADLE with a change of slope similar to that of DPDPE. These findings demonstrate a mixed mu-delta analgesic activity for the compound DADLE, which is often referred to as a prototypic delta agonist. These interactions differ from prior reports of none or minimal mu-ligand interactions with DADLE. Despite the cross-reactivity of DADLE to mu receptors, DADLE remains a more effective analgesic than do mu agonists in states of mu receptor tolerance.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3027303 TI - Ouabain binding to brain (Na+,K+)-adenosine triphosphatase: interactions of K+, ethanol and norepinephrine with high- and low-affinity binding. AB - Effects of K+, ethanol and norepinephrine on the binding kinetics of ouabain to (Na+,K+)-adenosine triphosphatase in beef brain microsomes were examined. K+ reduced the rate and apparent affinity for ouabain binding markedly. Whereas ethanol and norepinephrine themselves inhibited ouabain binding slightly, they stimulated binding in the presence of K+. Norepinephrine enhanced the effect of ethanol. Dissociation of ouabain was biphasic, with fast and slow components corresponding to high and low apparent affinity. About 65% of the enzyme had high affinity, regardless of conditions. Norepinephrine and ethanol had differential effects on the rate of dissociation from high and low affinity enzyme, however. Alpha receptor blockade generally prevented the effects of norepinephrine. These results show that, although norepinephrine and ethanol have a modest effect on the amount of enzyme that can bind ouabain, their main effect on (Na+,K+) adenosine triphosphatase is to antagonize the binding of K+ to its allosteric site that inhibits ouabain binding. The data support the hypothesis that ouabain binds rapidly to a K+-insensitive form of phosphorylenzyme or to its dephosphorylated analog and dissociates rapidly from E1. PMID- 3027304 TI - Regulation of beta adrenergic receptors on rat mononuclear leukocytes by stress: receptor redistribution and down-regulation are altered with aging. AB - In vitro incubation of mononuclear leukocytes (MNL) with catecholamines desensitizes beta adrenergic receptors, meaning isoproterenol-stimulated cyclic AMP accumulation decreases. This desensitization is accompanied by two patterns of receptor changes: first, reduction of surface receptors (defined as binding of [3H]dihydroalprenolol inhibited by 1 microM CGP 12177 [4-3-tertiarybutylamino-2 hydroxypropoxy)-benzimidazole- 2-on-hydrochloride]) without any change in the total number of [3H] dihydroalprenolol binding sites inhibited by 1 microM propranolol (receptor redistribution); then reduction of the total number of receptors (receptor down-regulation). In the present study we investigated receptor redistribution and down-regulation under physiological conditions by raising endogenous catecholamines in the rat by stress. In young rats a single immobilization stress induced MNL beta adrenergic receptor redistribution: the number of surface receptors was reduced by about 50% but the total number remained the same. Receptor redistribution was prevented completely in rats pretreated with beta-blocking nadolol. Repeated stress down-regulated the MNL beta adrenergic receptors as shown by a reduction in the total number of sites. We also investigated the regulation of beta adrenergic receptors in three age groups. After 60 min of immobilization stress the number of MNL surface receptors was reduced in young (4-month-old) rats but not in mature (12-month-old) or aged (26-month-old) rats. Using an alternative stress procedure, after single or repeated open-field sessions, we found receptor redistribution and down regulation, respectively, in young rats. None of these adaptive receptor response was observed in 26-month-old rats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3027305 TI - Differential effects of the imidazole derivatives etomidate, ketoconazole and miconazole and of metyrapone on the secretion of cortisol and its precursors by human adrenocortical cells. AB - The direct effects of etomidate, ketoconazole, miconazole and metyrapone were investigated on the secretion of cortisol and its precursors by dispersed cells from the adrenal cortex of a normal individual and four patients with Cushing's syndrome. The drugs interfered with adrenocorticotropic hormone-stimulated cortisol secretion in a dose-dependent way. Desoxycortisol concentrations in the medium were increased after the addition of metyrapone and low doses of etomidate but were suppressed with higher doses of etomidate. The production of 17-hydroxy progesterone was stimulated by miconazole and metyrapone but was strongly suppressed by the high doses of etomidate. Ketoconazole caused a stimulation of progesterone release, whereas the release of 17-hydroxyprogesterone was suppressed. Finally, the concentration of progesterone was strongly enhanced with high doses of miconazole, whereas etomidate suppressed progesterone production. It is concluded that etomidate at a low concentration (IC50, 10(-8) M) inhibits 11 beta-hydroxylase, whereas, at higher concentrations (10(-7)-10(-6) M), the side-chain cleavage enzyme system is also inhibited; metyrapone is a weaker (IC50, 10(-7) M) but pure 11 beta-hydroxylase inhibitor; miconazole inhibits adrenal 21-hydroxylase at 10(-6) M; and ketoconazole inhibits 17-hydroxylase. Etomidate, ketoconazole, miconazole and metyrapone inhibit cortisol biosynthesis in the human adrenal gland in different manners, which appear to involve the four cytochrome P-450-dependent monooxygenase reactions. Interestingly, these drugs affect corticosteroidogenesis by normal, hyperplastic and adenomatous adrenal cells in a similar manner. PMID- 3027306 TI - Presynaptic effects of l-alanine on peripheral sympathetic neurotransmission. AB - Concentrations ranging from 0.1 nM to 30 microM of the aminoacids l-alanine (l ALA), taurine, hypotaurine and beta-ALA were assayed on the chronotropic responses of rat isolated atria stimulated through their cardioaccelerans nerves. The increases in atrial rate obtained in frequency-response curves to nerve stimulation (0.1-12.8 Hz) were not modified by taurine, hypotaurine and beta-ALA but were reduced by l-ALA (0.01 and 0.1 microM). L-ALA did not modify the chronotropic responses to exogenous norepinephrine (NE). In the atria labeled in vitro with [3H]NE, the release of radioactivity evoked by postganglionic nerve stimulation at 2 Hz (6 V, 0.5 msec duration for 2 min) was reduced to 70% of control values by 0.1 microM l-ALA. The release of [3H]NE evoked by 60 and 100 mM KCl was also reduced by 0.1 microM l-ALA to 40 and to 70% of control values, respectively. L-ALA (0.1 microM) did not affect the outflow of tritium provoked by a 1-min incubation with 1 microM tyramine. The chronotropic responses to nerve stimulation were not modified by the l-ALA metabolites pyruvate and glutamate but were reduced by an analog of l-ALA, alpha-(methylamino)-isobutyric acid, which also diminished the release of [3H]NE elicited by nerve stimulation at 2 Hz.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3027307 TI - Evidence for the involvement of presynaptic alpha-2 adrenoceptors in the regulation of norepinephrine metabolism in the rat brain. AB - In an attempt to evaluate the possible functional role of alpha-2 adrenoceptors located on noradrenergic nerve endings in the regulation of cerebral norepinephrine metabolism, we have measured the effects of clonidine and idazoxan on cerebral free 3,4-dihydroxyphenylethyleneglycol (DOPEG) levels (an index of norepinephrine turnover) in the rat after surgical and experimental manipulations that allow an exclusive interaction of the alpha-2 adrenergic agents with presynaptic alpha-2 autoreceptors. The possible contribution of distant (to cell bodies) transsynaptic feedback mechanisms triggered by stimulation of postsynaptic alpha-2 adrenoceptors and of somatodendritic alpha-2 autoreceptor mediated regulatory mechanisms was eliminated by a local infusion of tetrodotoxin (50 ng) into the ascending noradrenergic bundle followed by electrical stimulation (at a frequency of 8 Hz) of this pathway distally to the neurotoxin injection site in chloral hydrate-anesthetized rats. Under these conditions, systemic injection of idazoxan (20 mg/kg i.p.) and clonidine (0.3 mg/kg i.p.) provoked an increase and a decrease, respectively, in free DOPEG levels in the hypothalamus, cerebral cortex and medial septum which were similar to those measured in naive rats. Moreover, in these animals the effect of idazoxan (1 mg/kg i.p.) was surmounted by a large dose of clonidine (0.3 mg/kg i.p.). The possible contribution of feedback mechanisms triggered by activation of postsynaptic alpha-2 adrenoceptors and mediated via local (to terminals) circuits (or a putative humoral agent released postsynaptically) was eliminated subsequently by a local injection of ibotenic acid in noradrenergic projection areas. Systemic administration of idazoxan (20 mg/kg i.p.) to ibotenate-lesioned rats elicited an increase in septal- and hypothalamic-free DOPEG levels comparable to that found in sham-operated rats. The effectiveness of the lesion was attested by a massive neuronal depopulation in the lesioned areas. Finally, ibotenic acid-induced destruction of noradrenergic target cells and local infusion of tetrodotoxin into followed by electrical stimulation of the ascending noradrenergic pathways were combined. Under these conditions, idazoxan still increased hypothalamic- and septal-free DOPEG levels, the extent of this alteration being similar to that found in normal rats. Altogether, these results suggest that irrespective of their low density, presynaptic alpha-2 autoreceptors play a cardinal role in the regulation of central nervous system norepinephrine metabolism. PMID- 3027308 TI - Rabbit myometrial adrenergic sensitivity is increased by estrogen but is independent of changes in alpha adrenoceptor concentration. AB - The ability of estrogen to increase the alpha-1 adrenergic sensitivity of rabbit uterine smooth muscle was compared with its effects on the concentration of alpha adrenoceptor subtypes in myometrium. Compared to ovariectomized controls, estrogen treatment was found to increase the adrenergic contractile sensitivity of rabbit uterus determined in vitro. Estrogen treatment increased uterine alpha 2 adrenoceptor concentration nearly 5-fold. Mature rabbits (endogenous estrogen) had the same uterine sensitivity and alpha-2 adrenoceptor concentration as estrogen-treated rabbits. Alpha-1 receptor concentration was increased only in the estrogen-treated group, and was accompanied by increased maximal contractile response. Thus, the increase in adrenergic sensitivity after estrogen was correlated closely with an increased concentration of alpha-2 adrenoceptors. However, enhanced adrenergic sensitivity persisted after alpha-1 and alpha-2 receptor concentration returned to pre-estrogen levels in rabbits pretreated with estrogen before ovariectomy. Studies utilizing subtype-selective competitive antagonists verified that contractile response is mediated primarily by alpha-1 adrenoceptors, with no apparent influence of alpha-2 adrenoceptors. Finally, we found that adrenergic sensitivity is altered by temperature and divalent cation concentration, but these effects do not prevent the expression of the regulatory action of estradiol. We conclude that estrogen increases the alpha-1 adrenergic sensitivity of rabbit uterus without changes in alpha-1 receptors, and thus may act on postreceptor response events.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3027309 TI - Diethylaminoethoxyhexestrol inhibition of purified rat liver lysosomal phospholipase A1: role of drug binding to substrate. AB - In this report, the inhibition of purified rat liver phospholipase A1 by diethylaminoethoxyhexestrol (DH) has been evaluated and the results correlated with DH binding to sonicated vesicles of di[1-14C]oleoylphosphatidylcholine. The drug bound in a positive cooperative manner to two classes of binding sites on phosphatidylcholine small unilamellar vesicles, one having an apparent high affinity and low capacity and another having a low affinity and high capacity. The observed data were shown to fit to a mixed type of inhibition when the free DH concentration (determined independently in the binding experiments) was used instead of the total drug concentration. Hydrolysis of enzyme-substrate-drug complexes was estimated to occur at a rate only half that of the enzyme-substrate complex. Results with DH suggest that both drug-enzyme and drug-substrate interactions may be important factors in the inhibition of lysosomal phospholipase A1. PMID- 3027310 TI - Evidence for diminished inhibitory neuromuscular transmission in distal small intestine. AB - The influence of myenteric nerves on duodenal muscle contraction and relaxation was examined in vitro and compared with muscle responses from the ileum. Proximal and distal rat small intestine was cut into strips measuring 6.0 X 10.0 mm. Strips cut along the oral-caudal axis were called longitudinal strips, whereas those cut 90 degrees to that axis were called circular strips. The strips were stretched to their optimal lengths and subjected to electrical field stimulation (0.1-1.0 msec pulse duration, 30-300 mA, 2-26 Hz) in the presence of Krebs' solution and Krebs' plus atropine, 10(-6) M. Field stimulation produced contraction responses that were inhibited by atropine and relaxation responses that were augmented by atropine. Muscarinic blockade abolished completely contraction in circular muscle, but atropine-resistant contractions persisted in the longitudinal strips. Proximal muscle showed significantly greater relaxation responses compared to distal muscle (P less than .05) at nearly all parameters of pulse duration, current and frequency. Contraction and relaxation amplitudes were significantly greater in longitudinal than respective circular muscle (P less than .05) at either site in the intestine. Thus, not only do the two muscle layers differ in their respective nerve supplies, but inhibitory neuromuscular transmission appears to have a greater influence in proximal than distal intestine. PMID- 3027311 TI - Alpha adrenergic receptors mediate angiotensin-induced prostaglandin production in the rabbit isolated vas deferens. AB - The effect of alpha adrenergic receptor antagonists on concentration-dependent response to angiotensins II and III was examined in the electrically stimulated isolated rabbit vas deferens. The force generated by a nonadrenergic neural mechanism was reduced by both peptides whereas the force generated by adrenergic neural mechanisms was enhanced. Angiotensin III-induced inhibition of the nonadrenergic contraction was significantly greater than that of angiotensin II for all groups. Yohimbine (1 X 10(-4) M), an alpha-2 receptor antagonist, attenuated the depression of the nonadrenergic contraction produced by angiotensins II and III. Yohimbine (1 X 10(-5) and 1 X 10(-4) M) also significantly reduced angiotensin II-induced prostaglandin E (PGE) synthesis. Yohimbine only significantly altered the angiotensin III-induced PGE synthesis at an antagonist concentration of 1 X 10(-4) M. Rauwolscine (1 X 10(-8) and 1 X 10( 7) M) attenuated angiotensin II-induced PGE production and at a higher concentration (1 X 10(-6) M) reduced angiotensin III-induced PGE production. The alpha-1 antagonist, prazosin (1 X 10(-6) M), did not alter nonadrenergic contractile or PGE responses to either angiotensin. The alpha-2 agonist, clonidine, both inhibited the nonadrenergic neural contraction and enhanced PGE synthesis. We interpret these data to indicate that angiotensins II and III may act via separate mechanisms to induce PGE synthesis in the vas deferens, with angiotensin II effects being more dependent on norepinephrine release from adrenergic nerves. PMID- 3027312 TI - Comparative effects of divalent cations on the methylmercury-induced alterations of acetylcholine release. AB - Bath application of methylmercury (MeHg) at the murine neuromuscular junction blocks synchronous evoked release of acetylcholine (ACh) and then increases spontaneous release of ACh effects observed electrophysiologically as cessation of EPPS, and increased MEPP frequency (MEPPf), respectively. The objectives of the present study were to test whether the effect of MeHg on spontaneous release was Ca++-specific by substituting Sr++ or Ba++ for Ca++, whether the time course of MeHg-induced block of synchronous evoked release was altered by varying Ca++ concentrations or substituting Sr++ and whether the processes involved in the decay of elevated MEPPf after repetitive stimulation (asynchronous evoked release) were altered by MeHg. MEPPf was recorded continuously from the rat hemidiaphragm using conventional methods during pretreatment with 2 mM Ca++, 2 mM Sr++ or 0.5 mM Ba++ and subsequently with the cation plus 100 microM MeHg. The time to peak MEPPf in MeHg was not different under any condition; however, peak MEPPf was lower in Sr++ solutions than in Ca++ or Ba++ solutions. EPPs were recorded from the rat hemidiaphragm cut muscle preparation during pretreatment with either 2, 4 or 8 mM Ca++ or 2 or 4 mM Sr++ and subsequently with the cation plus 100 microM MeHg. The latency to block of the EPP in 4 and 8 mM Ca++ was not significantly different from the latency in 2 mM Ca++. The latency to block in 2 or 4 mM Sr++ was also not different from that in Ca++. In addition, under all conditions EPP amplitude remained virtually unchanged from pretreatment values until block occurred after 8 to 9 min exposure to MeHg.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3027313 TI - Alpha adrenergic responses of blood vessels of rabbits after ovariectomy and administration of 17 beta-estradiol. AB - Experiments were designed to determine the effect of estrogen pretreatment on alpha adrenergic responsiveness of blood vessels of the rabbit. Rabbits were ovariectomized and, after 8 days of recovery, treated with 17 beta-estradiol (100 micrograms i.m.; estrogen group) or solvent (control group) for 4 days. Rings of saphenous vein and femoral artery (both without endothelium) were mounted for isometric tension recording in organ chambers filled with modified Krebs-Ringer bicarbonate solution (37 degrees C), gassed with 95% O2-5% CO2. All experiments were performed in the presence of inhibitors of neuronal uptake, extraneuronal uptake and beta adrenoceptors. In the saphenous vein, the estrogen treatment did not significantly affect the concentration-effect curves evoked by norepinephrine (either under control conditions or after alpha-1 or alpha-2 adrenergic blockade), phenylephrine (an alpha-1 adrenergic agonist) or UK 14,304 (an alpha-2 adrenergic agonist). In the femoral artery, estrogen treatment depressed the contractile responses evoked by norepinephrine (under control conditions) but not those produced by phenylephrine; UK 14,304 did not evoke a contractile response. The depressant effect of estrogen treatment on the concentration-effect curve to norepinephrine in the femoral artery was prevented by the alpha-2 adrenergic antagonist, rauwolscine. The results in the femoral artery but not in the saphenous vein suggest that estrogens depress alpha-2 but not alpha-1 adrenergic responsiveness. In the femoral artery, alpha-2 adrenoceptor stimulation does not cause contraction per se but apparently can facilitate alpha-1 adrenergic responses. This probably results from a reduced density of alpha-2 adrenoceptors in this blood vessel.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3027314 TI - Localization of sites in periventricular forebrain mediating cardiovascular effects of gamma-aminobutyric acid agonists and antagonists in anesthetized cats. AB - Previous studies have shown that injection of gamma-aminobutyric acid (GABA) antagonists such as bicuculline methiodide (BMI) into the forebrain (i.e., lateral and third) ventricular system elicits neurally mediated increases in blood pressure and heart rate and inhibits baroreflex bradycardia in anesthetized cats. Similar administration of muscimol, a GABA agonist, blocks or reverses the effects of BMI but has no effect on blood pressure or heart rate in untreated animals. These findings suggest that GABAergic inhibition may tonically suppress a forebrain mechanism capable of modifying autonomic outflow to the cardiovascular system. In the present study, we used a technique designed to restrict the distribution of intraventricularly administered drugs to varying degrees in order to better localize the relevant sites of drug action. Our findings show that BMI increases heart rate and blood pressure and that muscimol counters these changes by acting at a site that is accessible from the intermediate (as opposed to the rostral or caudal) region of the third ventricle. In contrast, these agents influence baroreflex bradycardia by acting at more rostral periventricular sites. These findings are consistent with the notion that the GABAergic mechanisms involved in the cardiovascular effects resulting from intraventricular administration of BMI and muscimol are located in the periventricular hypothalamus. PMID- 3027315 TI - Receptor reserve at the alpha-2 adrenergic receptor in the rat cerebral cortex. AB - N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ), an irreversible alpha-2 antagonist, was used to establish and quantitate the receptor reserve at the alpha-2 adrenergic autoreceptor mediating inhibition of [3H]norepinephrine ([3H]NE) release in rat cerebral cortical slices. EEDQ treatment had no effect on [3H]NE uptake or base-line release. Four hours after EEDQ treatment (0.8 mg/kg i.p.), the EC50 was shifted 7-fold to the right and there was a 21.5% decrease in the maximal response to the full alpha-2 agonist UK-14304. Using the double reciprocal plot analysis, the equilibrium activation constant (KA) was calculated to be 1.41 +/- 0.8 microM. Similar analysis of alpha-2 autoreceptor response at various times after 1.6 mg/kg of EEDQ gave similar values for the KA. Therefore, evaluation of either the response of the remaining native receptors after partial irreversible inactivation or the response of newly synthesized receptors after nearly complete irreversible inactivation can be used to determine the KA of the receptor. Comparison of repopulation kinetics analyses for alpha-2 receptor response and estimated receptor number revealed that recovery of maximal response was much faster than actual receptor recovery. By examining the relationship between alpha-2 autoreceptor occupancy and response it was possible to determine that there is approximately a 60 to 70% receptor reserve; only 1.5% of the receptors need to be occupied by UK-14304 in order to obtain 50% of the maximal inhibition of [3H]NE release. The presence of a large receptor reserve must be taken into account when evaluating alpha-2 adrenergic autoreceptor regulation in the rat cerebral cortex. PMID- 3027316 TI - Benzodiazepine receptor occupancy in vivo: correlation with brain concentrations and pharmacodynamic actions. AB - A tracer radioligand technique employing [3H]Ro 15-1788 was used to measure in vivo benzodiazepine receptor occupancy in mice. Animals were administered clonazepam (CNZ) or lorazepam (LRZ) i.p. or i.v., followed by [3H]Ro 15-1788. Plasma and cerebral cortical concentrations of the drug and cortical benzodiazepine receptor occupancy were determined at varying doses and times after administration. For both drugs, plasma and brain concentrations decreased in parallel with decreasing doses and with time, and the brain/plasma ratios remained constant. Receptor occupancy was maximal for CNZ and LRZ at doses above 1 and 4 mg/kg, respectively, and decreased with decreasing doses. Comparison of receptor occupancy with brain concentration yielded IC50 values of 21 ng/g for CNZ and 133 ng/g for LRZ. The apparent dissociation constant (Kd) for CNZ was 56 pmol/g and, for LRZ, 372 pmol/g. Hill plots yielded slopes of 1.30 for CNZ and 1.71 for LRZ. Alterations in the nonspecific uptake or elimination of Ro 15-1788 did not explain occupancy changes, inasmuch as brain concentrations of unlabeled Ro 15-1788 (6 mg/kg) were not changed by LRZ administration. The correlation between receptor occupancy and the pharmacodynamic actions of these drugs in blocking pentylenetetrazol seizures and inducing rotarod ataxia indicated that ED50 for these effects occurred at receptor occupancy of 30 to 60% for both drugs. PMID- 3027317 TI - An analysis of naltrexone precipitated abstinence in morphine-dependent chronic spinal dogs. AB - Graded doses of naltrexone (0.31, 1.125, 5.0, 20.0 and 80.0 micrograms/kg) were administered to five beagle-type dogs dependent on increasingly large stabilization doses of morphine (0.5, 1.0, 2.0, 4.0, 8.0, 16.0 and 24.0 mg/kg/day) and the intensity of precipitated abstinence (PAS) was determined by a previously developed scoring system. A complete crossover design was executed with all dogs receiving all doses of both naltrexone and morphine. The data were analyzed using a two-way analysis of variance (dogs and treatments). Treatment variance was partitioned into a regression and residual component. A mathematical model for relating the intensity of abstinence (PAS) to the concentrations of morphine and naltrexone was developed using the law of mass action and assuming that the degree of morphine physical dependence is related directly to the number of mu receptors occupied by morphine and the intensity of PAS is proportional to the number of receptors from which morphine is displaced by the antagonist. Using the mathematical model the deviations of the observed values from the calculated values were minimized using an iterative curve-fitting procedure which varied the values of the dissociation constants of morphine (KA) and naltrexone (KB) and the activity coefficient for morphine (alpha). The analysis of variance showed that the treatment effect was significant and that the regression accounted for most of this variance. The values which provided the best fit were: KA, 1.58 mg/kg; KB, 0.95 microgram/kg; and alpha, 245.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3027318 TI - Evidence for mu opioid receptor mediation of enkephalin-induced electroencephalographic seizures. AB - The opioid receptor types involved in the mediation of enkephalin-induced electroencephalographic (EEG) seizures were studied in unanesthetized, freely moving rats. Four receptor-selective peptide ligands were evaluated for effectiveness in producing nonconvulsive EEG seizures after i.c.v. administration; these included the mu agonist, [D-Ala2-N-methyl-Phe4-Gly5 ol]enkephalin (DAGO), the mixed mu-delta agonist, [D-Ala2-D-Leu5]enkephalin (DADLE), and the selective delta agonists, [D-Pen2-D-Pen5]enkephalin and [D-Pen2 L-Pen5]enkephalin. Only DAGO and DADLE were found to produce EEG seizures, with DAGO being 9 times more potent than DADLE. DAGO produced a greater number of seizure episodes with a greater overall incidence compared with DADLE, reflecting its potent effect to elicit EEG seizure activity in these rats. Injections of [D Pen2-D-Pen5]enkephalin or [D-Pen2-L-Pen5]enkephalin, even at the highest doses tested, failed to produce seizure activity. Behaviorally, the DAGO and DADLE EEG seizures were nonconvulsive but were temporally associated with episodic bursts of wet-dog shakes. The enkephalin-induced responses were extremely sensitive to antagonism by naloxone and completely blocked by pretreatment with the irreversible mu antagonist beta-funaltrexamine. The selective delta opioid receptor antagonist ICI 174,864 (N,N-diallyl-Tyr-Aib-Aib-Phe-Leu-OH) was ineffective. The use of the most selective agonists and antagonists for mu and delta opioid receptors suggests that, in rats, enkephalin-induced EEG seizures are mediated exclusively by mu opioid receptors and not by delta opioid systems. PMID- 3027319 TI - Denervation augments alpha-2 but not alpha-1 adrenergic responses in canine saphenous veins. AB - Experiments were performed in order to determine the influence of sympathetic denervation on alpha-1 and alpha-2 adrenergic responses in canine saphenous veins. In female dogs anesthetized with sodium pentobarbital, the left lumbar sympathetic chain was excised from L1 to L7. After a 3- to 5-week period, the left (denervated) and right (innervated) saphenous veins were removed, cut into rings and suspended for isometric tension recording in organ chambers filled with modified Krebs-Ringer bicarbonate solution. Denervation reduced significantly the norepinephrine content of the venous rings and the contractile responses evoked by the indirect sympathomimetic amine, tyramine. The contractile responses evoked by exogenous norepinephrine were augmented by denervation under control conditions (16.7-fold shift in concentration-effect curve) and also after inhibition of neuronal and extraneuronal uptake and beta adrenoceptors (3.8-fold shift in curve). Denervation increased the contractile responses evoked by the alpha-2 adrenergic agonist, UK 14,304 (5-fold shift in concentration-effect curve), but not those produced by the alpha-1 adrenergic agonist, phenylephrine. The selective augmentation of alpha-2 adrenergic responses by denervation may reflect the preferential innervation of alpha-2 adrenoceptors in the canine saphenous vein. PMID- 3027320 TI - Effect of phenobarbital on the distribution of drug metabolizing enzymes between periportal and perivenous rat hepatocytes prepared by digitonin-collagenase liver perfusion. AB - Intact periportal (pp) or perivenous (pv) hepatocytes were prepared by digitonin collagenase liver perfusion. The degree of separation was indicated by significant differences between the pp and pv cells in their activity of the pp markers, alanine aminotransferase (pp/pv = 2.1), gamma-glutamyltranspeptidase (3.4) and lactate dehydrogenase (1.3), and of the pv markers, glutamate dehydrogenase (0.73) and pyruvate kinase (0.81). This pattern was not altered by a 3-day pretreatment with phenobarbital (PB). The hepatocytes isolated from the pv area contained higher activities of microsomal NADPH-cytochrome c reductase, 7 ethoxycoumarin O-deethylase, 7-ethoxyresorufin O-deethylase and benzo(a)pyrene hydroxylase, and of cytosolic glutathione transferase. Cytochrome P-450 and UDP glucuronosyltransferase were slightly higher in pv cells. Treatment with PB induced NADPH-cytochrome c reductase, glutathione transferase, cytochrome P-450 and UDP-glucuronosyltransferase but the degree of induction was found to be at least as strong in pp cells as in pv cells. The induction of 7-ethoxyresorufin O deethylase and 7-ethoxycoumarin O-deethylase was clearly more prominent in pp cells. On the other hand, PB reduced the activities of benzo(a)pyrene hydroxylase and alcohol dehydrogenase in both cell types. These results demonstrate by direct enzyme assay of separated cells the dominance of the pv-region for metabolizing drugs in the normal liver. Contrary to several other studies, however, our data indicate that induction by PB occurs panacinarily, i.e., relatively more in the pp region, thus diminishing rather than exaggerating the original pv dominance. PMID- 3027321 TI - [Lipiodolized angiography in hepatocellular carcinomas. Contribution of iodine 131-labelled lipiodol]. AB - Twelve patients with hepatocellular carcinoma and cirrhosis were investigated by Lipiodol injection into the hepatic artery. A CT scan was done 4-6 days later. Lipiodol was retained by hepatic tumors in each case. This method emphasized the extension of the carcinoma and allowed to discover daughter tumors. I131-lipiodol was also injected in 4 of the 12 patients and then its biodistribution was evaluated. At the 6th hour after injection, I131-lipiodol was detected by scintigraphy over the liver (74-91 percent) and over the lungs (9-16 percent) only. The tumor to normal liver pixel count ratio was about 5. These results indicate that there is a preferential arterial blood flow towards the hepatic tumors, and that we can consider a therapeutic use of I131-Lipiodol in hepatocellular-carcinoma. PMID- 3027323 TI - Rapid recovery of gonadotroph function after down-regulation of receptors for GnRH in ewes. AB - Several characteristics of the hypothalamo-hypophysial axis were examined after down-regulation of GnRH receptors and the desensitization which accompanies it in the ewe. Down-regulation of GnRH receptors, induced by i.v. infusion of GnRH (2.5 micrograms/h) for 24 h, resulted in a 50% decrease in the number of receptors for GnRH at the end of the infusion period. The number of receptors for GnRH was restored to control values by 6 h after the infusion ended and remained stable at 12, 24, 48, 72 and 96 h after infusion. The amount of LH released in response to an i.v. injection of 100 micrograms GnRH was reduced by 82% at the end of the infusion period, but there was no significant reduction in the GnRH-induced release of FSH. The GnRH-induced release of LH was restored by 12 h after the infusion ended; however, the amount of FSH released in response to GnRH was not different from control values at any time. A decrease in both the amplitude and frequency of endogenous pulses of LH was observed from 0 to 12 h after the end of the infusion period. At no time did the concentration of gonadotrophins in the pituitary change. These results demonstrate that replenishment of receptors for GnRH and recovery of the ability of the gonadotroph to release LH are associated events. However, the GnRH-induced release of FSH does not appear to be closely related to the number of GnRH receptors. We suggest that continuous exposure to GnRH may inhibit the hypothalamic pulse generator as well as the pituitary response to the pulse generator. PMID- 3027322 TI - Effects of long-term treatment with melatonin or melatonin plus ectopic pituitary transplants on testicular LH/hCG and prolactin receptors in juvenile Syrian hamsters (Mesocricetus auratus). AB - Juvenile hamsters were injected daily with melatonin and some were also given transplants of 2 pituitaries under the kidney capsule. Weights of the testes and the accessory reproductive glands were reduced after 8 and after 12 weeks of melatonin treatment, but remained unaltered in animals treated with ectopic pituitary transplants. Levels of testicular LH/hCG receptors were significantly reduced by daily melatonin injections for 8 and 12 weeks. The presence of pituitary transplants in melatonin-injected hamsters prevented these reductions, and increased LH/hCG receptors above control levels. These changes in testicular LH/hCG receptors were closely related to alterations in serum prolactin concentration induced by melatonin and pituitary transplants. After 8, but not after 12 weeks of treatment, testicular prolactin receptor levels were reduced by melatonin and maintained by the presence of pituitary transplants. We conclude that: juvenile male hamsters become sensitive to the effects of daily melatonin injections when they reach maturity; daily melatonin injections can reduce the levels of testicular LH/hCG and prolactin receptors; and the effects of melatonin on LH/hCG and prolactin receptors are probably due to suppression of endogenous prolactin release. PMID- 3027324 TI - Hammond memorial lecture. New concepts of the control of corpus luteum function. AB - Five new concepts concerning the control of corpus luteum function in the cow have been developed in recent years. Prostacyclin (PGI-2) plays a luteotrophic role. Conversely, products of the lipoxygenase pathway of arachidonic acid metabolism, particularly 5 hydroxyeicosatetraenoic acid (5-HETE), play luteolytic roles. Luteal cells arise from two sources. The small luteal cells are all of theca cell origin; the large cells found early in the cycle (Days 2-6) are mainly of granulosa cell origin. However, a population of large cells found after Day 10 of the cycle are of theca cell origin. Oxytocin of luteal cell origin plays a role in development of the corpus luteum and possibly in its regression. The recently described Ca2+-polyphosphoinositol-protein kinase C second messenger system, as well as the LH-cAMP system, is involved in control of progesterone synthesis in the bovine corpus luteum. Progesterone synthesis in the small theca derived luteal cells is primarily controlled by the cAMP system. However, elevated intracellular calcium diminishes cAMP-mediated progesterone synthesis in these cells. These findings modify our current concepts of the mechanisms of control of progesterone secretion by the corpus luteum and suggest several new lines of research. PMID- 3027325 TI - Fetal ventriculomegaly and brain atrophy in a woman with intrauterine cytomegalovirus infection. A case report. AB - Fetal ventriculomegaly and brain atrophy occurred in a woman with intrauterine cytomegalovirus infection. PMID- 3027327 TI - Spin probes as mechanistic inhibitors and active site probes of thymidylate synthetase. AB - C-4- and C-5-substituted analogues of dUMP were examined as inhibitors of thymidylate synthetase and as topographical probes of its active site by electron spin resonance (ESR). The C-5-substituted spin-labeled analogues pDUAP (2) and a pDUTT (3) as well as the unlabeled AAdUMP (1) were competitive inhibitors with Ki's of 9.2, 89, and 7.9 microM, respectively. The C-4-spin-labeled pls4dU (4) displayed no inhibition activity. Scatchard plots as determined by ESR gave similar association constants for 2 (Kassoc = 1.9 X 10(5) M-1) and for 3 (Kassoc = 2.4 X 10(5) M-1). Both of these values are similar to the Kassoc of FdUMP indicating that the bulky substituent in position 5 does not interfere with the formation of the binary complex. The enzyme-C-5-spin-labeled nucleotide complexes indicate the presence of similarly immobilized spin labels by ESR, whereas no binding and immobilization were noticed with the C-4-spin-labeled nucleotide. A model for the active-site geometry of the enzyme was derived which suggests that the C-5 substituents point toward the opening of the binding cavity whose depth is at least 12 A. Also, the approximate 10-fold increased inhibitory activity of 2 as compared to that of 3 may be attributed to the significant electron withdrawing properties of the C-5 substituent in 2. Finally, the set of probes used for the binding and inhibition of thymidylate synthetase gives direct experimental evidence that an electron-withdrawing C-5 substituent primarily affects the formation of the ternary complex and will not substantially influence the stability of the binary complex. PMID- 3027326 TI - Structure-activity relationship of novel oligopeptide antiviral and antitumor agents related to netropsin and distamycin. AB - A group of oligopeptides have been synthesized that are structurally related to the natural antiviral antitumor agents netropsin and distamycin. Cytostatic activity against both human and murine tumor cell lines as well as their in vitro activity against a range of viruses is reported. The biological activity of these agents is discussed both in terms of their structural differences and, in particular, in relation to their observed base- and sequence-dependent minor groove binding to duplex oligonucleotides. PMID- 3027328 TI - Synthesis and antitumor and antiviral properties of 5-halo- and 5 (trifluoromethyl)-2'-deoxyuridine 3',5'-cyclic monophosphates and neutral triesters. AB - The title diesters (11-15; halo substituents F, Cl, Br, I) were prepared by DCC induced cyclization of the precursor 5'-monophosphate or direct halogenation of the 2'-deoxyuridine 3',5'-cyclic monophosphate. Antitumor activities of 11-15 in cell systems (L1210 and Raji/0) were compared to those of the corresponding nucleosides and 5'-monophosphates. Thus, the 5-F- and 5-CF3-2'-deoxyuridines proved to be highly active derivatives [ID50 values (microgram/mL) for L1210, 0.002 and 0.06, respectively], with the 5'-monophosphates showing comparable potencies. The corresponding 3',5'-cyclic monophosphate diesters were 20-30 times less potent but nonetheless highly cytostatic. All derivatives including 11-15 had greatly increased ID50 values for the thymidine kinase deficient (TK-) L1210 and Raji cells. The 3',5'-cyclic diesters (11-15) evidently are not efficient prodrug sources of the nucleoside 5'-monophosphates in TK- cells. They also proved to be 100- to 2000-fold less efficient inhibitors of L1210 thymidylate synthetase than were the 5'-monophosphates. The 5-substituted 2'-deoxyuridines and their 5'-monophosphates were potent inhibitors of herpes simplex virus (MIC50 mostly 0.07-10 micrograms/mL) and vaccinia virus (MIC50 0.07-0.2 microgram/mL), with antiviral activity decreasing in the order 5-I, 5-Br greater than 5-CF3 greater than 5-Cl greater than 5-F. The 3',5'-cyclic monophosphates (11-15) were for the most part 10- to 40-fold less active than the 5'-monophosphates in the virus assay systems (e.g., MIC50 for the 5-Br and 5-I derivatives ranged 1-20 micrograms/mL). By contrast 11-15 were considerably more potent inhibitors of vaccinia virus growth (MIC50 0.4-2 micrograms/mL). As the neutral 3',5'-cyclic methyl phosphate triesters (16-18), the 5-I and 5-Br compounds were less potent in antiviral and cytostatic agents than the 3',5'-cyclic diesters, while the 5 iodo benzyl triester was in several cases as active as the 3',5'-cyclic diester. The title compounds (11-15) appear to require extracellular hydrolysis to the nucleoside before functioning as antitumor or antiviral agents. PMID- 3027329 TI - Synthesis and biological activity of 5-phenylselenenyl-substituted pyrimidine nucleosides. AB - Several 5-phenylselenenyl derivatives of pyrimidine nucleosides were synthesized by electrophilic addition of phenylselenenyl chloride to the nucleosides under basic conditions. With use of this route, 5-(phenylselenenyl)-6-azauracil was also prepared. These compounds may serve as inhibitors of thymidylate synthase, as potential antiviral and anticancer agents, and as versatile intermediates for the synthesis of 5- or 6-substituted nucleosides. 5 (Phenylselenenyl)arabinosyluracil (PSAU, 4) and the corresponding cytosine analogue (PSAC, 5) were poor inhibitors of a promyelocytic leukemia cell line that was arabinosylcytosine-resistant. PSAU and PSAC were significantly less active than ara-C against L1210 cells and were found to selectively interfere with the cellular uptake and/or phosphorylation of 2'-deoxycytidine and 2' deoxyuridine in intact L1210 cells. PMID- 3027330 TI - Structure-activity relationships of kadsurenone analogues. AB - Kadsurenone, a specific receptor antagonist of platelet-activating factor (PAF), and its analogues were prepared from derivatives of cinnamyl alcohol and (allyloxy)phenol. Racemic kadsurenone, resolvable by a Chiralpak column at low temperatures, has an IC50 value of 2 X 10(-7) M, which is about 50% of the activity of the natural product (IC50 = 1 X 10(-7) M). The structural specificity of kadsurenone was further demonstrated by the low PAF-receptor-blocking activities of denudatin B, mirandin A, desallylkadsurenone, and the 2-epimer of kadsurenone. PMID- 3027332 TI - Substituted (aryloxy)alkanoic acids as antagonists of slow-reacting substance of anaphylaxis. AB - A series of compounds in which the 4-acetyl-3-hydroxy-2-propylphenoxy moiety of the standard SRS-A antagonist, FPL-55712, is linked by a polymethylene or a polyether chain to substituted (aryloxy)alkanoic acids was prepared. The compounds were evaluated for their ability to antagonize SRS-A-induced contractions of guinea pig ilea and LTE-induced bronchoconstriction in the guinea pig. The results showed that the compounds were all less potent than FPL-55712 in vitro, yet surprisingly, most were more potent by the inhalation route of administration. Some of the most potent analogues were selected for further pharmacological evaluation and, by inhalation, exhibited selective antagonism of leukotrienes as compared with PAF or histamine. In comparison to FPL-55712, compounds 28 and 37 were more potent against LTE (40- and 80-fold, respectively), LTD (4- and 3-fold, respectively), and LTC (27- and 20-fold, respectively) induced bronchoconstriction when tested by inhalation. PMID- 3027331 TI - Preparation, receptor binding, and fluorescence properties of hexestrol fluorophore conjugates: evaluation of site of attachment, fluorophore structure, and fluorophore-ligand spacing. AB - We have undertaken a staged development of certain estrogen-fluorophore conjugates, in order to prepare a fluorescent estrogen suitable for determination of the estrogen receptor content of individual cells. Since non-steroidal estrogens with bulky substituents are often more readily bound by receptor than their steroidal counterparts, we have investigated fluorophore conjugates with derivatives of the non-steroidal estrogen hexestrol [3R*, 4S*)-3,4-bis(4 hydroxyphenyl)hexane). On the basis of the receptor-binding affinity of model compounds, we prepared a prototypical set of three ring- and side-chain substituted fluorescent hexestrol derivatives, whose binding and fluorescence properties ultimately led to the preparation of a series of side-chain substituted nitrobenzoxadiazole derivatives. The compounds prepared have binding affinities for the estrogen receptors that range from ca. 1% to 5% that of estradiol, and they have very favorable fluorescence characteristics, similar to those of fluorescein. PMID- 3027333 TI - Synthesis and biological evaluation of acyclic neplanocin analogues. AB - Acyclic neplanocin analogues were prepared by condensation of adenine or N2 acetylguanine with (E)-1,4-dichlorobut-2-ene and subsequent hydrolysis. The N-9 substituted product 9-[(E)-4-hydroxybut-2-enyl]adenine was obtained when adenine was employed as the starting purine, while N2-acetylguanine yielded both the N-7 and N-9 isomers. Cell-culture studies revealed that only the chloro-substituted intermediate 9-[(E)-4-chlorobut-2-enyl]adenine exhibited significant cytotoxicity against P-388 mouse lymphoid leukemia cells, while the N-9-substituted guanine analogue 9-[(E)-4-hydroxybut-2-enyl]guanine inhibited replication of herpes simplex viruses type 1 and type 2. PMID- 3027334 TI - Nucleosides. 139. Synthesis and anticytomegalovirus and antiherpes simplex virus activity of 5'-modified analogues of 2'-fluoroarabinosylpyrimidine nucleosides. AB - In order to determine if modification of the 5'-position reduces or abolishes the antiviral activity of 2'-fluoro-5-iodo-ara-C (FIAC), 2'-fluoro-5-iodo-ara-U (FIAU), or 2'-fluoro-5-methyl-ara-U (FMAU) against human cytomegalovirus (HCMV) and herpes simplex virus (HSV), the 5'-deoxy, 5'-mercapto, and 5'-amino analogues of these nucleosides were prepared. 5'-Deoxy-FIAC and 5'-deoxy-FIAU were prepared by catalytic hydrogenation of 5'-iodo-FIAC and 5'-iodo-FIAU to 5'-deoxy-FAC and 5'-deoxy-FAU, respectively, followed by reiodination at C-5. Reduction of 5'-iodo FMAU afforded 5'-deoxy-FMAU. These 5'-deoxy nucleosides were found to be inactive against HCMV, indicating that the conversion to 5'-phosphate by the cellular enzyme(s) is a requirement for antiviral activity against this virus. Other 5' modified (NH2 and SH) analogues were also prepared from 5'-O-tosyl-FIAC and 5'-O tosyl-FMAU. Treatment of these tosylates with LiN3 in DMF afforded the corresponding 5'-N3 products. Catalytic hydrogenation of 5'-N3-FMAU afforded 5' NH2-FMAU, whereas 5'-NH2-FIAC was obtained by treatment of 5'-N3-FIAC with Ph3P in pyridine. 5'-Mercapto analogues were prepared by treatment of 5'-O-tosyl-3'-O acetyl nucleosides with KSAc followed by deacetylation. 5'-NH2-FMAU was the only compound that showed good activity against HSV-1 and HSV-2 in vitro. However, this compound was less potent and had a lower therapeutic index than FMAU. PMID- 3027335 TI - In search of the digitalis replacement. AB - There are now several new CT drugs undergoing clinical study for the treatment of CHF. While most of these new agents are like digitalis in that they possess significant inotropic activity, they differ from digitalis in that they tend to also have pronounced vasodilator properties. Such compounds, because they are not pure CT agents, should not be regarded as true digitalis replacements. Despite the increased understanding about the cAMP cascade and about cardiac muscle contraction in general, the long search for a specific digitalis replacement cannot be regarded as having been accomplished. It should also be noted that the initial clinical assessment for any of these new compounds will be complicated by the fact that they are tested in only the latter stages of CHF where prognosis is very poor and the possibility of showing significant improvement in mortality is extremely difficult. It will be unfortunate if the potential for positive inotropic agents (pure CT drugs) is compromised by clinical studies that employ mixed inotropic-vasodilator compounds in extremely sick patient populations. The continued use of the digitalis glycosides for such a long period of history implies that at least some subpopulation of CHF patients should derive benefit from therapy with an appropriately selective CT drug. Neither the appropriate drug nor the appropriate specific patient population, both of which are needed to properly address the eventual role of inotropic therapy in CHF, have, as yet, been identified. PMID- 3027336 TI - Bimorphinans as highly selective, potent kappa opioid receptor antagonists. PMID- 3027337 TI - Synthesis and structure-activity relationships of a series of aminopyridazine derivatives of gamma-aminobutyric acid acting as selective GABA-A antagonists. AB - We have recently shown that an arylaminopyridazine derivative of GABA, SR 95103 [2-(3-carboxypropyl)-3-amino-4-methyl-6-phenylpyridazinium chloride], is a selective and competitive GABA-A receptor antagonist. In order to further explore the structural requirements for GABA receptor affinity, we synthesized a series of 38 compounds by attaching various pyridazinic structures to GABA or GABA-like side chains. Most of the compounds displaced [3H]GABA from rat brain membranes. All the active compounds antagonized the GABA-elicited enhancement of [3H]diazepam binding, strongly suggesting that all these compounds are GABA-A receptor antagonists. None of the compounds that displaced [3H]GABA from rat brain membranes interacted with other GABA recognition sites (GABA-B receptor, GABA uptake binding site, glutamate decarboxylase, GABA-transaminase). They did not interact with the Cl- ionophore associated with the GABA-A receptor and did not interact with the benzodiazepine, strychnine, and glutamate binding sites. Thus, these compounds appear to be specific GABA-A receptor antagonists. In terms of structure-activity, it can be concluded that a GABA moiety bearing a positive charge is necessary for optimal GABA-A receptor recognition. Additional binding sites are tolerated only if they are part of a charge-delocalized amidinic or guanidinic system. If this delocalization is achieved by linking a butyric acid moiety to the N(2) nitrogen of a 3-aminopyridazine, GABA-antagonistic character is produced. The highest potency (approximately equal to 250 times bicuculline) was observed when an aromatic pi system, bearing electron-donating substituents, was present on the 6-position of the pyridazine ring. PMID- 3027338 TI - Inhibitors of cyclic AMP phosphodiesterase. 1. Analogues of cilostamide and anagrelide. AB - Evaluation of a series of lactam heterocyclic analogues of cilostamide (2) as inhibitors of cyclic AMP phosphodiesterase derived from both human platelets and rat heart in comparison with their corresponding methoxy-substituted heterocycles has revealed that the N-cyclohexyl-N-methyl-4-oxybutyramide side chain of 2 is an important lipophilic and/or steric pharmacophore. Attachment of this side chain to the parent heterocycle of the potent cyclic AMP phosphodiesterase inhibitor anagrelide (3) afforded the hybrid structure RS-82856 (1), shown to be more potent than either of its progenitors as an inhibitor of cyclic AMP phosphodiesterase or of ADP-induced platelet aggregation. The available in vitro data suggest that 1 possesses potentially useful antithrombotic and cardiotonic properties. PMID- 3027339 TI - Inhibitors of cyclic AMP phosphodiesterase. 2. Structural variations of N cyclohexyl-N-methyl-4-[(1,2,3,5-tetrahydro- 2-oxoimidazo[2,1-b]quinazolin-7-yl) oxy]butyramide (RS-82856). AB - A series of analogues of the cyclic AMP phosphodiesterase (PDE) inhibitor N cyclohexyl-N-methyl-4-[(1,2,3,5-tetrahydro- 2-oxoimidazo[2,1-b]quinazolin-7 yl)oxy]butyramide (RS-82856, 1) was prepared by systematic variation of the side chain substituent, length, position, connecting atom, and the parent heterocycle itself. The compounds were evaluated as inhibitors of cyclic AMP phosphodiesterase from both human platelets and rat or dog heart tissue and as inhibitors of ADP-induced platelet aggregation. Structure-activity correlations for the analogue series revealed significant limitations on the steric bulk of substituents on the 1,2,3,5-tetrahydroimidazo[2,1-b]quinazolin-2-one heterocycle and the position and length of the side chain. As inhibitors of cyclic AMP phosphodiesterase (PDE), potency steadily increased with increasingly lipophilic side chains. In platelet aggregation inhibition studies, however, a maximum in activity was reached with 1, while more lipophilic compounds were significantly less active. Major changes in the heterocycle itself, represented by isomeric and other carbonyl variations, also decreased activity. The molecular features defined by this series of analogues of 1 correlate to a high degree with current understanding of the chemical and topographical requirements of the active site of the FIII (type IV) form of cyclic AMP PDE. Selective inhibition of this enzyme has been proposed as the principal component of the positive inotropic action of a number of cardiotonic agents. PMID- 3027340 TI - Structure-activity studies of 5-[[4-(4,5-dihydro-2-oxazolyl) phenoxy]alkyl]-3 methylisoxazoles: inhibitors of picornavirus uncoating. AB - A series of substituted phenyl analogues of 5-[[4-(4,5-dihydro-2-oxazolyl) phenoxy]alkyl]-3-methylisoxazoles has been synthesized and evaluated in vitro against several human rhinovirus (HRV) serotypes. Substituents in the 2-position greatly enhanced activity when compared to the unsubstituted compound. Many of these compounds exhibited mean MICs (MIC) against five serotypes as low as 0.40 microM. The mean MIC correlated well (r = 0.83) with the MIC80 (the concentration that inhibited 80% of the serotypes tested). A quantitative structure-activity relationship study indicated a strong dependency of MIC on lipophilicity (log P) in combination with inductive effects (sigma m) and bulk factors (MW). PMID- 3027341 TI - 5-Quinone derivatives of 2'-deoxyuridine 5'-phosphate: inhibition and inactivation of thymidylate synthase, antitumor cell, and antiviral studies. AB - Both photochemical aromatic substitution and palladium (0)-catalyzed biaryl coupling reactions have been employed in the synthesis of 5-substituted 2' deoxyuridines. The former procedure was useful in the preparation of the 3,4 dimethyl-2,5-dimethoxyphenyl derivative 12a and the 3,4,6-trimethyl-2,5 dimethoxyphenyl derivative 12b. The latter reaction was efficient in the preparation of the 2-(3-methyl-1,4-dimethoxynaphthyl) derivative 14. These compounds and their nucleotides (20a-c) were converted to the corresponding quinone nucleosides 19a-c and nucleotides 6-8 by an oxidative demethylation reaction using ceric ammonium nitrate and silver(II) oxide, respectively. The kinetics and products of the reaction of the quinone nucleosides 19a,b with methyl thioglycolate showed rapid addition to the quinone ring in the trisubstituted derivative 19a and somewhat slower redox reactions with the tetrasubstituted quinones 19b and 19c. All six nucleotides had high affinity for the title enzyme from Lactobacillus casei with Ki values ranging from 0.59 to 3.6 microM; the most effective compounds were the dimethyl quinone 6 and the naphthoquinone 8. Somewhat higher inhibitory constants were observed with the quinones against the L1210 enzyme. The dimethyl quinone nucleotide 6 showed time dependent inactivation (kinact = 0.015 s-1) against the L. casei enzyme, a rate saturation effect, and substrate protection in accord with the kinetic expression for an active-site-directed alkylating agent. The apparent second-order rate of this reaction (2.5 X 10(4) M-1 s-1) is one-twentieth the rate (kcat.) of the normal enzymatic reaction leading to product. None of the compound exhibited sufficient activity in the antitumor cell or antiviral assays to warrant further study. PMID- 3027342 TI - 5-Amino-4-(diazoacetyl)-1-beta-D-ribofuranosylimidazole, a new antileukemic agent. AB - 5-Amino-4-(diazoacetyl)-1-beta-D-ribofuranosylimidazole (15), 5-amino-4 (chloroacetyl)-1-beta-D-ribofuranosylimidazole (16), and a number of related imidazole ribonucleosides have been synthesized. Compounds 15 and 16 are cytotoxic to both H.Ep.-2 and L1210 leukemia cells in culture. The (diazoacetyl)imidazole 15 is also active against the P388 leukemia in mice. PMID- 3027344 TI - Immunological studies on Crohn's disease. VI. Increased chemiluminescent response of peripheral blood monocytes. AB - Oxidative metabolic activity can be measured by a luminol-dependent chemiluminescence (CL) assay. To investigate the phagocytic activity of blood cells, CL responses were assessed in 26 patients with Crohn's disease (CD) and 26 healthy controls, using heparinized peripheral blood. Significantly (p less than 0.005) increased CL responses of whole blood cells were found in 26 patients with CD. However, CL activity was not directly related to disease activity nor to other clinical parameters. The CL responses did not correlate with the number of white blood cells, neutrophils nor monocytes. In consideration of the increased CL, there is the possibility that functional abnormalities in some components of blood cells leads to elevations in CL responses of whole blood cells. To obtain support for this proposal, blood cells were separated into neutrophils and monocytes, and CL of neutrophils or monocytes were studied, using the same number of each type of cells as that of the controls. The oxidative metabolism of monocytes from CD patients was significantly (p less than 0.005) higher than that of controls, while the phagocytic activity of neutrophils was similar to that in the controls. These data suggest that elevations in CL activity of whole blood cells were not due to alterations of the neutrophils but rather to abnormalities in the monocytes. Further attention directed to the significance of monocyte function in the pathogenesis of CD is warranted. PMID- 3027343 TI - Duchenne muscular dystrophy with adrenal insufficiency and glycerol kinase deficiency: high resolution cytogenetic analysis with molecular, biochemical, and clinical studies. AB - A syndrome of Duchenne muscular dystrophy (DMD), adrenal hypoplasia, glycerol kinase deficiency, and mental retardation has been recognised. We report a further case ascertained from a history of DMD, severe mental retardation, and an Addison-like disorder. Cytogenetic analysis of the proband revealed an interstitial deletion of the short arm of the X chromosome. from Xp21.1 to Xp22.11, comprising about 9% of the length of the normal X chromosome. His mother was heterozygous for the deletion, but his maternal grandmother and sister both had two normal X chromosomes. DNA probe analysis confirmed the existence of a deletion in the affected boy, as probes 754, C7, XJ1-1, and pERT87 consistently failed to hybridize to his DNA. His sister was heterozygous for the RFLP associated with 754, thus confirming that she had two normal X chromosomes. There was no evidence of chronic granulomatous disease, other immunological defect, or retinitis pigmentosa in this case. Biochemical studies revealed gross glyceroluria and hyperglycerolaemia, indicating glycerol kinase deficiency which has been confirmed enzymatically. We have subsequently screened 21 other boys with DMD for glyceroluria and found one other case. Cytogenetic analysis has also been performed in nine other families, where a boy with DMD has been shown to have a deletion of DNA sequences localised to the region Xp21. None of these cases demonstrated any cytogenetic abnormality, nor has their clinical course been unusual. PMID- 3027346 TI - Congenital anomalies in two neonatal tamarins (Saguinus oedipus and Saguinus fuscicollis). AB - Congenital anomalies were seen at necropsy of two neonatal tamarins. The defects included achondroplastic-like dwarfism, polydactyly and syndactyly in a Saguinus oedipus, and scoliosis and uterus didelphys in a S. fuscicollis. Both infants were the offspring of incestuous matings between twin siblings. PMID- 3027345 TI - Defective granulocyte chemotaxis and natural killer activity in a patient with recurrent infections. AB - A 25-year-old male with a history of recurrent infections presented with fever, severe aplastic anaemia, splenomegaly and retroperitoneal node enlargement. Lympho-histiocytic granulomas were found in spleen, liver and lymph nodes. Granulocyte studies revealed normal morphology, severely impaired random migration and complete absence of directed locomotion. Whereas phagocytosis was slightly reduced, candidacidal activity and nitroblue tetrazolium reduction were normal. Basal granulocyte cyclic GMP levels were within the normal range while a 5-fold increase of cyclic AMP levels was observed. Numerous abnormalities were also found in the patient's lymphocytes: lack of delayed hypersensitivity, reduced response to mitogens, low OKT4/OKT8 ratio, absence of natural killer (NK) activity with normal number of cells recognized by NK-specific monoclonal antibodies. These observations describe a distinct clinico-pathologic entity and suggest the possibility of a common defect in granulocytes and in NK cells. PMID- 3027347 TI - Protein kinase C activates the renal apical membrane Na+/H+ exchanger. AB - Studies were performed on purified brush-border membranes from the kidney of the rabbit to examine the relation between protein kinase C and the Na+/H+ exchanger in these membranes. The brush-border membranes were transiently opened by exposure to hypotonic media and the membrane proteins phosphorylated by exposure to ATP and phorbol esters or partially purified protein kinase C. The membranes were resealed and the intravesicular space acidified by incubation in a sodium free isotonic solution (pH 5.5). The rate of uptake of 1 mM 22Na+ (pH 7.5), with and without amiloride (1 mM), was assayed and the proton gradient-stimulated, amiloride-inhibitable component of 22Na+ taken as a measure of the activity of the Na+/H+ exchanger. 12-0-tetradecanoyl phorbol-13-acetate (TPA) increased the amiloride-sensitive component of 22Na+ uptake. TPA did not affect the amiloride insensitive component of 22Na+ uptake or the equilibrium concentration of sodium. TPA also did not affect the rate of dissipation of the proton gradient in the absence of sodium or the rate of sodium-dependent or -independent uptake of D glucose. Other "active" phorbol esters stimulated the rate of Na+/H+ exchange, but phorbol esters of the 4 alpha configuration did not. Incubation of the opened membranes in partially purified protein kinase C increased the rate of proton gradient-stimulated, amiloride-inhibitable sodium uptake. The stimulatory effect of TPA and protein kinase C was not additive. In the absence of ATP, neither TPA nor protein kinase C affected Na+/H+ exchange transport. To determine the membrane-bound protein substrates, parallel experiments were conducted with gamma [32P] ATP in the phosphorylating solutions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3027348 TI - Zinc inhibition of potassium efflux in depolarized frog muscle and its modification by external hydrogen ions and diethylpyrocarbonate treatment. AB - Efflux of 42K+ was measured in frog sartorius muscles equilibrated in hyperosmotic depolarizing solutions. At the internal potentials obtained, K+ passes mainly through the inward rectifier potassium channels. Inhibition of K+ efflux by external Zn2+ (0.25 to 15 mM) differs in three significant ways from inhibition by Ba2+. (1) The dose-response relation does not correspond to action at a single site. (2) The Zn2+-sensitivity of K+ efflux does not depend on [K+]0 at constant internal potential. (3) Zn2+ inhibition is reduced by hydrogen ions, while Ba2+ inhibition is unaffected. Further, the Ba2+-sensitivity of K+ efflux is not altered by a half-inhibiting Zn2+ concentration, suggesting that the two ions do not interact at a common site. The histidine-modifying reagent diethylpyrocarbonate (DEPC) reduces Zn2+ inhibition. After DEPC treatment Zn2+ inhibition is further reduced by low pH. DEPC has little effect on Ba2+ inhibition. Zn2+ inhibition is not altered by treatment with the sulfhydryl reagents 5,5'-dithio-bis(2-nitrobenzoic acid) or dithiothreitol. The results can be described by either of two models in which two sites can bind Zn2+ and one or both of the sites may also bind H+. When both sites bind Zn2+, K+ efflux is inhibited, and a third site may then bind H+. The effects of DEPC can be accounted for by a decrease in H+ affinity of the first two sites by a factor of 50, and a decrease in Zn2+ affinity of these sites and of the H+ affinity of the third site by about one order of magnitude. PMID- 3027349 TI - Topoisomerase II cleavage in chromatin. AB - We have examined the effect of the anti-tumor drug VM-26 on purified Drosophila topoisomerase II, and used this drug to map (putative) topoisomerase II cleavage sites in chromatin. These studies indicate that VM-26 interferes with the strand breakage-rejoining catalytic cycle. VM-26 appears to stabilize the topoisomerase II-cleavable complex and markedly enhances the formation of double-strand breaks in naked DNA. VM-26 also stimulates the formation of double-strand breaks in isolated Drosophila nuclei. Analysis of the parameters of the VM-26-stimulated cleavage reaction in nuclei strongly suggests that the double-strand scissions are generated by endogenous topoisomerase II. Finally, we have examined the distribution of (putative) cleavage sites for endogenous topoisomerase II in the chromatin of the 87A7 heat shock locus and the histone repeat unit. We have found that there are prominent VM-26-induced cleavage products from the 5' ends of the 87A7, the two heat shock protein 70 genes, and in the intergenic spacer separating these genes. Moreover, the pattern of VM-26-induced cleavage products is altered in nuclei prepared from heat-shocked cells. In the case of the histone repeat unit, only minor VM-26-induced cleavage products are observed in nuclei (in spite of the fact that experiments on naked DNA indicate that the histone repeat contains many major cleavage sites for purified topoisomerase II). These findings suggest that the nucleoprotein organization of different DNA segments may be important in determining whether specific sites are accessible to endogenous topoisomerase II in nuclei. PMID- 3027350 TI - Alpha adrenoceptors and the myocardium. I.S.H.R. (International Society for Heart Research) 12th congress. 9-13th February, 1986. Melbourne. PMID- 3027351 TI - Dual trophic effects of the alpha 1-adrenergic receptor in cultured neonatal rat heart muscle cells. AB - The molecular signals regulating myocardial hypertrophy are unknown. We used a new model system to study this problem. Muscle cells from the neonatal rat ventricle were cultured in serum-free medium. Control cells did not change in size over time and did not show spontaneous contractile activity. Incubation with norepinephrine or epinephrine had two major trophic effects that developed over 1 2 days. The first was stimulation of muscle cell hypertrophy or increase in size. The second was induction of spontaneous contractile activity. The hypertrophic response was mediated through an alpha 1-adrenoceptor. The beating response required both alpha 1- and beta 1-adrenergic receptor stimulation. Alpha 1 stimulation alone produced enlarged cells that did not beat. Alpha 1 plus beta 1 stimulation induced contractile activity even when protein synthesis and hypertrophy were inhibited. Thus, the two alpha 1 responses could be dissociated, providing evidence for two independently-regulated cellular pathways for the dual alpha 1 trophic effects. One pathway leads to hypertrophy. The other, which requires concomitant beta 1 stimulation, controls the development of beating. Preliminary work raises the possibility that different products of inositol phospholipid hydrolysis might initiate the two alpha 1 trophic pathways. Other preliminary work suggests that the growth pathway is associated with the expression of specific genes and might reflect an action of the alpha 1 adrenoceptor on the cell nucleus. These studies define previously unsuspected trophic roles for the alpha 1-adrenergic receptor. PMID- 3027352 TI - Myocardial alpha-adrenoceptors and positive inotropy. AB - The positive inotropic effect (PIE) of beta-adrenoceptor stimulating agents in the mammalian heart is mainly due to an increase in slow Ca++ inward current (Isi) which in turn is probably the result of an increase in intracellular cAMP levels. The present paper is concerned with the alpha(probably alpha 1) adrenoceptor-mediated PIE which differs from the response to beta-adrenoceptor stimulation in several points: it develops relatively slowly, is not accompanied by an abbreviation but instead by a prolongation of the contraction, is dependent on the frequency of stimulation, is increased in hypothyroidism and is not accompanied by detectable changes in cAMP and cGMP levels. In spite of the failure to elevate cAMP levels, alpha-adrenoceptor stimulation increases the magnitude, and decelerates the decay, of Isi. The alpha-adrenoceptor mediated increase in Isi is smaller than that of an equieffective (with respect to the PIE) concentration of isoprenaline. However, this effect conceivably contributes to the alpha-adrenoceptor-mediated PIE although other mechanisms (e.g. an increase in Ca++ sensitivity of the contractile proteins) are likely to be also involved. Recent experiments provide evidence that blockade of beta-adrenoceptors increases the density of myocardial alpha 1-adrenoceptors. This is in accord with the view that stimulation of alpha-adrenoceptors by endogenous catecholamines may serve as a reserve mechanism under conditions of impaired beta-adrenergic influence. PMID- 3027353 TI - Nucleotide sequence of the BamHI repetitive sequence, including the HindIII fundamental unit, as a possible mobile element from the Japanese monkey Macaca fuscata. AB - Clustered repeat units produced by BamHI digestion of genomic DNA from the Japanese monkey Macaca fuscata [JMr(BamHI)] were sequenced by dideoxy DNA sequencing. The nucleotide sequences of several individual repeats showed that the BamHI repeat contains the 170-bp HindIII element as an integral part, and that it has more than 90% homology with the HindIII repeat element [AGMr(HindIII)] found in the genomic DNA of the African green monkey. In the JMr(BamHI) repeat unit, the 170-bp HindIII element is flanked by a 6-bp inverted repeat, which is part of a 22-bp direct repeat. This latter repeat of 22-bp asymmetrically overlaps the border between the internal AGMr(HindIII)-like region and adjacent regions of the JMr(BamHI) repeat. A similar structural feature of the BamHI repeat unit has been found in the genomic DNA of the baboon, but not in that of the African green monkey. These results show clearly that the BamHI repeat of the modern Japanese monkey originated as a result of insertion of an AGMr(HindIII) element into a certain site(s) of the genomic DNA of an ancestor of the modern Japanese monkey before Macaca-Cercocebus divergence. PMID- 3027354 TI - Evolutionary change of restriction sites under unequal rates of nucleotide substitution among the three positions of codons. AB - Under the assumption of unequal rates of nucleotide substitution among the three positions of codons, mathematical formulas are derived for the probability that a restriction site observed at time 0 in a DNA sequence will be present at time t, the probability that a new restriction site will emerge at a particular place in two descendant sequences, and the proportion of identical restriction sites between two such sequences. All three quantities are shown to be larger for the case of unequal rates than for the case of equal rates. As a consequence, estimates of nucleotide divergence (2 lambda t) between two sequences based on restriction-site data tend to be lower than the actual values if the assumption of equal rates is made but actually does not hold. However, the degree of underestimation is slight if 2 lambda t is 0.10 or smaller. The underestimation can be serious when 2 lambda t becomes 0.20 or larger, particularly if the substitution rates at the three codon positions are very different or if transitional substitution occurs more frequently than transversional substitution. The underestimation is more serious for four-base enzymes than for six-base enzymes. PMID- 3027356 TI - Hepatocellular carcinoma and lupoid hepatitis (an autopsy study). PMID- 3027355 TI - Sequences of four mouse histone H3 genes: implications for evolution of mouse histone genes. AB - The sequences of four histone H3 genes coding for the replication variant proteins H3.1 and H3.2 have been determined. Three of these genes, two coding for H3.1 proteins and one for an H3.2 protein, are located on chromosome 13 and expressed at low levels. The fourth gene, encoding an H3.2 protein, is located on chromosome 3 and expressed at a high level. The coding regions of the three genes on chromosome 13 are more similar to each other than to the H3 gene on chromosome 3, and equally divergent from it, suggesting that either gene duplication or gene conversion has occurred since the genes were dispersed onto two chromosomes. A 14 base sequence including the CCAAT sequence and located 5' to the genes on chromosome 13 has been conserved. The histone H3 gene on chromosome 3 has multiple potential binding sites for the Sp1 transcription factor. The coding regions show greater than 95% conservation among the four genes. This is due to the strict pattern of codon usage and the presence of two long (greater than 60 base) regions of completely conserved nucleic acid sequence. These conserved regions in the coding sequence may have an important functional role at the mRNA or DNA level. PMID- 3027357 TI - Sonography of gallbladder perforation: a complication of transcatheter hepatic arterial embolization. PMID- 3027358 TI - Phenotypic revertants of temperature-sensitive M protein mutants of vesicular stomatitis virus: sequence analysis and functional characterization. AB - Twenty-five spontaneous temperature-stable revertants of four different temperature-sensitive (ts) M protein mutants (complementation group III: tsG31, tsG33, tsO23, and tsO89) were sequenced and tested for their ability to inhibit vesicular stomatitis virus RNA polymerase activity in vitro. Consensus sequences of the coding region of each M protein gene were determined, using total viral RNA as template. Fifteen different sequences were found among the 25 revertants; 14 differed from their ts parent by a single amino acid (one nucleotide), and 1 differed by two amino acids (two nucleotides). Amino acids were altered in various positions between residues 64 and 215, representing over 60% of the polypeptide chain. Resequencing of the Glasgow and Orsay wild types and the four ts mutants confirmed previously published differences (Y. Gopalakrishana and J. Lenard, J. Virol., 56:655-659, 1985), and one or two additional differences were found in each. The relative charges of the revertant M proteins, as determined by nonequilibrium pH gradient electrophoresis, were consistent with the deduced sequences in every case. The ability of each revertant M protein to inhibit the RNA polymerase activity of nucleocapsids prepared from its parent ts mutant was also tested. Only 13 of the 25 revertants had M protein with high (wild type like) polymerase-inhibiting activity, while 5 had low (ts-like) activity, and 7 had intermediate activity, demonstrating that this property is not an essential concomitant of the temperature-stable phenotype. It is concluded that the high reversion frequency observed for these mutants arises from a very high incidence of pseudoreversion, i.e., many different molecular changes can repair the ts phenotype. PMID- 3027359 TI - Glucocorticoid regulation of murine leukemia virus transcription elements is specified by determinants within the viral enhancer region. AB - The transcriptional control region (the long terminal repeat, LTR) of the leukemogenic murine retrovirus SL3-3 contains a glucocorticoid-responsive consensus sequence, as does the corresponding region of the nonleukemogenic virus Akv. Dexamethasone increases gene expression directed by both LTR sequences. However, the responses of the LTRs of the two viruses to dexamethasone differ according to the cell line in which the response is measured. The results of these studies provide insights regarding differences in response to glucocorticoids dependent upon cell line and indicate that the glucocorticoid responsive elements may compose one of the determinants of tissue specificity and pathogenicity of the murine retroviruses. PMID- 3027360 TI - Stages in the nuclear association of the herpes simplex virus transcriptional activator protein ICP4. AB - The nuclear localization of the herpes simplex virus transcriptional activator protein ICP4 was studied by indirect immunofluorescence. At early times after viral infection, ICP4 quickly localized to a diffuse intranuclear distribution. ICP4 later concentrated in globular compartments within the nucleus. The redistribution to the compartments was dependent on viral DNA replication. Double staining for ICP4 and ICP8, the early major DNA-binding protein, revealed that both were found in the same intranuclear globular compartments at late times. These were previously named "replication compartments" (M. P. Quinlan, L. B. Chen, and D. M. Knipe, Cell 36:857-868, 1984). Because ICP4 and ICP8 are known to function in transcriptional activation and DNA replication, respectively, both DNA replication and late transcription may occur in these compartments. The association of ICP4 and ICP8 with the replication compartments appeared to be independent in that the retention of ICP8 in the compartments required ongoing viral DNA synthesis, while the association of ICP4 was independent of viral DNA synthesis once the compartments were formed. Because ICP4 shows a different distribution at early and late times, stimulation of transcription by ICP4 may involve different molecular events or contacts during these two periods of the replicative cycle. PMID- 3027361 TI - Expression of the Kirsten ras viral and human proteins in Escherichia coli. AB - The expression vectors pINIII-A and pINIII (lpp p5) were used to construct plasmids which direct the synthesis in Escherichia coli of the Kirsten ras viral (v-Ki-ras) and human cellular (c-Ki-ras) oncogene products as fusion proteins containing 9 and 10 extra amino acids, respectively, at their N termini. Authenticity of the bacterially produced proteins was determined by immunoprecipitation and immunoblot analyses with ras-specific monoclonal antibodies. After induction with isopropyl-beta-D-thiogalactopyranoside, the viral protein represented approximately 20% of the total cellular protein. The majority of the protein was found in the postsonication low-speed centrifugation pellet. The synthesized viral protein was active in GTP binding, as judged by autophosphorylation and photoaffinity labeling assays. PMID- 3027362 TI - Molecular characterization of the Akvr-1 restriction gene: a defective endogenous retrovirus-borne gene identical to Fv-4r. AB - A dominant restriction allele, Akvr-1r, from California wild mice (Mus musculus domesticus) confers resistance to exogenous ecotropic murine leukemia virus (MuLV) infection. The presence of an ecotropic MuLV envelope-related glycoprotein in uninfected virus-resistant cells suggests that viral interference is a possible mechanism for this resistance. We molecularly cloned the ecotropic MuLV envelope-related sequence from the genomic DNA of a wild mouse homozygous for the Akvr-1r locus. The cloned provirus was defective and contained a C-terminal end of the pol gene, a complete envelope gene, and a 3' long terminal repeat. The presence of this provirus was directly correlated with Akvr-1r-mediated virus resistance in cell cultures and hybrid mice. The Akvr-1r provirus restriction map and partial DNA sequence were identical to those of the Fv-4r allele, an ecotropic MuLV resistance locus from Japanese feral mice (M. musculus molossinus), which was previously shown to be allelic with the Akvr-1r gene. The 3' host flanking sequences of Fv-4r and Akvr-1r also had identical restriction maps. These findings indicate that Akvr-1r and Fv-4r are the same gene. It was probably acquired by interbreeding of these feral species in recent times. Conservation of this locus might be favored by the useful function that it performs in protection against ecotropic MuLV infection endemic in both populations of wild mice. PMID- 3027363 TI - Expression of cell-associated and secreted forms of herpes simplex virus type 1 glycoprotein gB in mammalian cells. AB - The gene for glycoprotein gB1 of herpes simplex virus type 1 strain Patton was expressed in stable Chinese hamster ovary cell lines. Expression vectors containing the dihydrofolate reductase (dhfr) cDNA plus the complete gB1 gene or a truncated gene lacking the 194 carboxyl-terminal amino acids of gB1 were transfected into CHO DHFR-deficient cells. Radioimmunoprecipitation demonstrated that the complete gB1 protein expressed in CHO cell lines was cell associated, whereas the truncated protein was secreted from the cells due to deletion of the transmembrane and C-terminal domains of gB1. Cells expressing the truncated gB1 protein were subjected to stepwise methotrexate selection, and a cell line was isolated in which the gB1 gene copy number had been amplified 10-fold and the level of expression of gB1 had increased over 60-fold. The truncated gB1 protein was purified from medium conditioned by the amplified cell line. N-terminal amino acid sequence analysis of this purified protein identified the signal peptide cleavage site and predicted the cleavage of a 30-amino-acid signal sequence from the primary protein. The immunogenicity of the truncated gB1 protein was also tested in mice, and high levels of antibody and protection from virus challenge were observed. PMID- 3027364 TI - Structure and expression of the herpes simplex virus type 2 glycoprotein gB gene. AB - The gene for glycoprotein gB2 of herpes simplex virus type 2 strain 333 was cloned, sequenced, and expressed in mammalian cells. The gB2 protein had an overall nucleotide and amino acid sequence homology of 86% with the cognate gB1 protein. However, of the 125 amino acid substitutions or deletions, only 12.5% were conservative replacements. These differences were clustered within an NH2 terminal region, a central region, and a COOH-terminal region, resulting in domains of near identity broken by small regions of marked divergence. Regions of greatest homology included a 90-amino-acid stretch starting at residue 484 and 39 amino acids spanning residues 835 to 873, which cover a rate-of-entry locus mapped to Ala-552 and a syn locus mapped to Arg-857, respectively, in gB1 by Bzik et al. (D. J. Bzik, B. A. Fox, N. A. DeLuca, and S. Person, Virology 133:301-314, 1984). Pellett et al. (P. E. Pellett, K. G. Kousoulas, L. Pereira, and B. Roizman, J. Virol. 53:243-253, 1985) mapped the mutations in three monoclonal antibody-resistant gB1 mutants between amino acids 273 and 443. These epitopes are included in a region of 98 residues identical between gB1 and gB2. The identity of this protein was verified by placing a truncated gene lacking the 303 carboxyl-terminal amino acids of gB2 into mammalian COS and CHO cells. Expression was demonstrated by immunofluorescence and radioimmunoprecipitation. This protein will be purified from the stable CHO cell lines and compared with gB1 for immunogenicity and protective efficacy in animal challenge models. PMID- 3027366 TI - Analysis of p60v-src mutants carrying lesions involved in temperature sensitivity. AB - Analysis of the src genes of three temperature-sensitive (ts) mutants of Rous sarcoma virus (tsNY68, tsNY72-4, and PA104) showed that each has two C-terminal mutations in the kinase domain required for temperature sensitivity, as assayed by morphological alteration and anchorage-independent growth. In all three mutants, one of the mutations is a valine-to-methionine change at position 461. To assess the contribution of each mutation to the biochemical properties of the src protein, we analyzed the kinase activity and the interaction with cellular proteins p50 and p90 of recombinant src gene products in which only one mutation was combined with wild-type src sequences. Chimeric src protein containing only the Met-461 mutation was indistinguishable from the wild type by all criteria examined, while the effect of the second C-terminal mutation alone varied with the defectiveness of the parental ts mutant. The second mutation alone, while not sufficient to cause ts transformation, altered p60src complex formation with cellular proteins p50 and p90 and altered the in vitro thermolability of src kinase activity. The results indicate that these biochemical properties of p60src are more sensitive to mutation than others, such as in vivo kinase activity, which require more profound structural alterations. PMID- 3027365 TI - Retrovirus integration and chromatin structure: Moloney murine leukemia proviral integration sites map near DNase I-hypersensitive sites. AB - The chromatin conformation of mouse genome regions containing Moloney murine leukemia proviral intergration sites in two Mov mouse strains and randomly selected integration sites in virus-infected mouse 3T3 fibroblasts was analyzed. All integrations have occurred into chromosomal regions containing several DNase hypersensitive sites, and invariably the proviral integration sites map within a few hundred base pairs of a DNase-hypersensitive site. The probability that this close association between proviral integration sites and DNase-hypersensitive sites was due to chance was calculated to be extremely low (2 X 10(-4]. Because the proviral integrations analyzed were not selected for an altered phenotype, our results suggest that DNase-hypersensitive regions are preferred targets for retrovirus integration. PMID- 3027367 TI - Octyl-beta-D-glucopyranoside extracts polyomavirus receptor moieties from the surfaces of mouse kidney cells. AB - Polyomavirus receptor moieties were extracted from the surfaces of mouse kidney cells with the nonionic detergent octyl-beta-D-glucopyranoside. Following extraction with this detergent, mouse kidney cells were refractory to polyomavirus infection. Binding studies demonstrated that this loss of susceptibility resulted from extraction of a peripheral membrane protein or proteins required for proper virus attachment to and infection of mouse kidney cells. Infection of extracted mouse kidney cells returned following a 2-h recovery period. However, the presence of cycloheximide or tunicamycin in the recovery media interfered with recovery from infection. Cells could be infected immediately after extraction by supplying them with the extracted moieties prior to or concomitant with infection. A complex of polyomavirus and the extracted receptor protein was formed by in vitro incubation and was stable in sucrose gradient analysis. Functional receptor moieties were prepared in the form of liposomes from the detergent extract. The virus-receptor complex was immunoprecipitated with anti-polyomavirus immunoglobulin G, and the portion of the complex contributed by the cell was identified. Immunoblot analysis of the mouse kidney cell detergent extract with a receptor-specific 125I-labeled anti idiotypic antibody or 125I-labeled polyomavirus demonstrated several reactive proteins. Attachment of polyomavirus to mouse kidney cells, followed by extraction of the virus-receptor complex, identified polyomavirus-binding proteins similar to those observed in in vitro binding. Proteins with molecular weights of approximately 95,000, 50,000 and 25,000 to 30,000 were consistently observed in all receptor assays. The relationship between these proteins and their possible involvement as the cell receptor for polyomavirus are discussed. PMID- 3027368 TI - Abrogation of resistance to severe mousepox in C57BL/6 mice infected with LP-BM5 murine leukemia viruses. AB - Strain C57BL/6 (B6) mice infected with LP-BM5 murine leukemia virus (MuLV) develop a disease which combines abnormal lymphoproliferation with profound immunosuppression and has many features in common with human acquired immunodeficiency syndrome induced by HTLV-III/LAV retroviruses. To determine whether this LP-BM5 MuLV infection would affect the innate resistance of B6 mice to a naturally occurring, highly virulent murine pathogen, mice were exposed to ectromelia virus at various times after treatment with LP-BM5 viruses. At week 4 after infection with LP-BM5, mice challenged with ectromelia virus were unable to generate a humoral immune response to this virus, and between weeks 8 and 10 after infection, challenged mice lost the ability to generate an ectromelia virus specific cytotoxic-T-cell response. Loss of the cellular immune responses to ectromelia virus was associated with an increased susceptibility to the lethal effects of the virus. PMID- 3027369 TI - A single-base change within the DNA polymerase locus of herpes simplex virus type 2 can confer resistance to aphidicolin. AB - An aphidicolin-resistant (Aphr) mutant of herpes simplex virus (HSV) type 2 strain 186 previously has been shown to induce an altered viral DNA polymerase that is more resistant to aphidicolin and more sensitive to phosphonoacetic acid (PAA) than is wild-type DNA polymerase. In this study the mutation responsible for the aphidicolin-resistant phenotype was physically mapped by marker transfer experiments. The physical map limits for the Aphr mutation were contained in a 1.1-kilobase pair region within the HSV DNA polymerase locus. The 1.1-kilobase pair fragment of the Aphr mutant also conferred hypersensitivity to PAA, and DNA sequence analysis revealed an AT to GC transition within this fragment of the Aphr mutant. Analysis of the three potential open reading frames within the 1,147 base-pair fragment and comparison with the amino acid sequence of DNA polymerase of HSV type 1 indicated that the Aphr mutant polymerase had an amino acid substitution from a tyrosine to a histidine in the well-conserved region of the DNA polymerase. These results indicate that this single amino acid change can confer altered sensitivity to aphidicolin and PAA and suggest that this region may form a domain that contains the binding sites for substrates, PPi, and aphidicolin. PMID- 3027370 TI - Medium tumor antigen of polyomavirus transformation-defective mutant NG59 is associated with 73-kilodalton heat shock protein. AB - Affinity-purified medium T antigen encoded by NG59, a nontransforming mutant of polyomavirus, is specifically associated with a protein of 72,000 daltons (72K protein). Medium T antigens of wild-type polyomavirus and the transformation competent mutant dl8 are not associated with the 72K protein. Instead, they form a complex with another protein of 61,000 daltons. Several lines of evidence suggest that the medium T antigen-associated 72K protein is equivalent to the abundant and constitutive 73K heat shock protein. First, on two-dimensional polyacrylamide gels the 72K protein migrated with the same pI (5.6) as did the 73K heat shock protein. Second, the 72K protein was immunoprecipitable with antibodies against heat shock proteins. Third, when digested with V8 protease, the 72K protein gave rise to the same pattern of fragments as did the 73K heat shock protein. PMID- 3027371 TI - Monoclonal antibodies as probes for a function of large T antigen during the elongation process of simian virus 40 DNA replication. AB - Various monoclonal antibodies specific for simian virus 40 large tumor antigen (T antigen) inhibit the elongation process of viral DNA replication in an in vitro system. The results provide strong evidence for a function intrinsic to T antigen during ongoing replicative-chain elongation. The antibody inhibition studies were further used to establish a correlation between the known biochemical activities of T antigen and its function during the elongation phase. The data demonstrate that, in addition to DNA binding and ATPase, a third function of T antigen is required for replicative chain elongation. This function is most probably related to the recently described DNA helicase activity of T antigen. This conclusion is based on the following results: aphidicolin treatment of actively replicating simian virus 40 minichromosomes causes a partial uncoupling of parental DNA strand separation and DNA synthesis; the strand separation reaction is blocked by the same monoclonal antibodies which strongly inhibit the elongation process. DNA helicase activity of isolated T antigen is equally well inhibited by the same set of monoclonal antibodies that affect minichromosome replication in vitro. PMID- 3027372 TI - Isolation and characterization of recombinants between attenuated and virulent aphthovirus strains. AB - A guanidine-resistant mutant of the attenuated strain of aphthovirus type 01 strain Campos and the original wild-type strain were crossed to generate recombinant viruses. Two independently derived recombinant viruses were isolated. One isolate (RI) contained the P1 (structural proteins) gene region of attenuated strain and P3 (polymerase precursor) gene region of the wild-type strain. The other isolate (RII) had a genomic structure complementary to that of RI, this is, P1 of the wild-type strain and P3 of the attenuated virus. Recombinant RII inherited some in vitro phenotypic markers that were characteristic of the attenuated strain, whereas the RI recombinant had in vitro behavior that was similar to that of the wild-type strain. The data obtained suggest that the polymerase precursor (P3) of the attenuated strain (01 Campos) could be involved in the determination of the attenuated phenotype for fetal bovine kidney cells and, eventually, for cattle. PMID- 3027374 TI - Primary structure and transcription of the genes coding for the two virion phosphoproteins pp65 and pp71 of human cytomegalovirus. AB - Human cytomegalovirus contains a phosphorylated matrix protein of 65,000 apparent molecular weight (65K phosphoprotein; pp65) and a related phosphoprotein of 71,000 molecular weight (pp71). The 65K phosphoprotein is usually by far the most abundant structural component found in culture-grown purified virus particles. This study describes the precise mapping of the genes for both polypeptides, giving the entire nucleotide sequences and the exact positions of the respective transcripts. The 65K phosphoprotein is coded for by the 5'-terminal part of an abundant 4-kilobase (kb) mRNA. The 71K phosphoprotein corresponds to the single translational reading frame of a rare nonspliced 1.9-kb mRNA that is coterminal with the 4-kb transcript. The promoter for 4-kb mRNA appears to be unusual in structure; it does not contain a characteristic TATA sequence. The expression of antigenic epitopes from pp65 may allow improved serodiagnosis of human cytomegalovirus infections. PMID- 3027373 TI - trans-dominant defective mutants of simian virus 40 T antigen. AB - We constructed a collection of linker insertion mutants in the simian virus 40 (SV40) genome and studied several of these with changes limited to a part of the large T antigen gene corresponding to an amino acid sequence shared with other ATPases. Two of these mutants were found to have a novel phenotype in that they could not be complemented for plaque formation by a late-region deletion mutant. These two mutants, in contrast to other mutants in this region, were able to transform rat cells in culture at a frequency close to that of the wild-type gene. The noncomplementing mutants were found to be potent inhibitors of SV40 DNA replication despite the presence of wild-type T antigen in the transfected cells. This inhibition was shown to be the result of the introduced mutations in the large T antigen gene. We conclude that the large T antigens of the noncomplementing mutants can act as inhibitors of SV40 DNA replication. PMID- 3027375 TI - Continuing coevolution of virus and defective interfering particles and of viral genome sequences during undiluted passages: virus mutants exhibiting nearly complete resistance to formerly dominant defective interfering particles. AB - We quantitatively analyzed the interference interactions between defective interfering (DI) particles and mutants of cloned vesicular stomatitis virus passaged undiluted hundreds of times in BHK-21 cells. DI particles which predominated at different times in these serial passages always interfered most strongly (and very efficiently) with virus isolated a number of passages before the isolation of the DI particles. Virus isolated at the same passage level as the predominant DI particles usually exhibited severalfold resistance to these DI particles. Virus mutants (Sdi- mutants) isolated during subsequent passages always showed increasing resistance to these DI particles, followed by decreasing resistance as new DI particles arose to predominate and exert their own selective pressures on the virus mutant population. It appears that such coevolution of virus and DI particle populations proceeds indefinitely through multiple cycles of selection of virus mutants resistant to a certain DI particle (or DI particle class), followed by mutants resistant to a newly predominant DI particle, etc. At the peak of resistance, virus mutants were isolated which were essentially completely resistant to a particular DI particle; i.e., they were several hundred thousand-fold resistant, and they formed plaques of normal size and numbers in the presence of extremely high multiplicities of the DI particle. However, they were sensitive to interference by other DI particles. Recurring population interactions of this kind can promote rapid virus evolution. Complete sequencing of the N (nucleocapsid) and NS (polymerase associated) genes of numerous Sdi- mutants collected at passage intervals showed very few changes in the NS protein, but the N gene gradually accumulated a series of stable nucleotide and amino acid substitutions, some of which correlated with extensive changes in the Sdi- phenotype. Likewise, the 5' termini (and their complementary plus-strand 3' termini) continued to accumulate extensive base substitutions which were strikingly confined to the first 47 nucleotides. We also observed addition and deletion mutations in noncoding regions of the viral genome at a level suggesting that they probably occur at a high frequency throughout the genome, but usually with lethal or debilitating consequences when they occur in coding regions. PMID- 3027376 TI - Epstein-Barr virus nuclear antigen forms a complex that binds with high concentration dependence to a single DNA-binding site. AB - A bacterially synthesized 28-kilodalton carboxyl-terminal fragment (28K-EBNA of Epstein-Barr virus nuclear antigen shows highly concentration dependent binding to monomer, dimer, and trimer copies of synthetic DNA-binding site 5' GATCTAGGATAGCATATGCTACCCCGGGG 3' 3' ATCCTATCGTATACGATGGGGCCCCCTAG 5' in bacterial plasmids. The rate of the binding reaction is independent of the number of sites, but dependent upon the length of the DNA containing the sites. These data are consistent with 28K-EBNA locating its binding sites by a process of facilitated transfer or sliding along the DNA. The highly concentration dependent binding suggests that multiple 28K-EBNA monomer polypeptides form a complex before or during binding. Binding occurs equally well at 24 and 37 degrees C, but not at 0 degrees C. A 28K-EBNA complex bound to a single site has unoccupied binding sites capable of interacting with additional DNA molecules. Such interaction is confirmed by agarose gel electrophoresis of protein-DNA complexes which indicate that a 28K-EBNA complex forms bridges between two DNA molecules. A bridge between the two binding regions in the Epstein-Barr virus origin of plasmid replication (oriP) would form a loop structure which could be an important feature for the regulatory function of authentic Epstein-Barr virus nuclear antigen. PMID- 3027377 TI - Complete nucleotide sequence of a milk-transmitted mouse mammary tumor virus: two frameshift suppression events are required for translation of gag and pol. AB - We sequenced two recombinant DNA clones constituting a single provirus of the milk-transmitted mouse mammary tumor virus characteristic of BR6 mice. The complete provirus is 9,901 base pairs long, flanked by 6 base-pair duplications of cellular DNA at the site of integration. Five extensive blocks of open reading frame corresponding to the gag gene, the presumed protease, the pol and env genes, and the open reading frame orf within the long terminal repeat of the provirus were readily discernible. Translation of gag, protease, and pol involved three different translational reading frames to produce the three overlapping polyprotein precursors Pr77, Pr110, and Pr160 found in virus-infected cells. Synthesis of the reverse transcriptase and endonuclease therefore required two separate frameshifts to suppress the termination codons at the ends of the Pr77 and Pr110 domains. Direct evidence is presented for translational readthrough of both stop codons in an in vitro protein synthesis system. PMID- 3027379 TI - Differences in biological activity and structural protein VP1 phosphorylation of polyomavirus progeny resulting from infection of primary mouse kidney and primary mouse embryo cell cultures. AB - Both primary mouse kidney and primary mouse embryo cells in culture were used for polyomavirus progeny production. Examination of polyomavirus virion structural integrity revealed that mouse embryo cell progeny contained a threefold greater population of unstable particles when compared with mouse kidney cell progeny. Differences in biological activity between these two progeny virion types were also shown. Mouse kidney cell progeny compared with mouse embryo cell progeny exhibited a 10-fold greater ability to agglutinate guinea pig erythrocytes, a 3 fold lower ability to become internalized into monopinocytotic vesicles, and a 2 fold lower ability to initiate a productive infection based on positive nuclear immunofluorescence when mouse embryo host cell cultures were used. The mouse kidney progeny were also found to bind to host cells less specifically than the mouse embryo cell progeny. When these two progeny virion types were labeled in vivo with 32P and subjected to isoelectric focusing followed by sodium dodecyl sulfate-polyacrylamide gel electrophroesis in the second dimension, differences in the phosphorylation pattern of the major virus-encoded structural protein VP1 species were observed. It was revealed that species D and E of mouse kidney cell progeny were phosphorylated to the same degree, while mouse embryo cell progeny species E and F were phosphorylated equally. These data suggest that the host cells play a role in modulating the biological activity of the virus by affecting the degree and site-specific phosphorylation of the major capsid protein VP1 which may influence the recognition of virus attachment proteins for specific cellular receptors. PMID- 3027378 TI - Epstein-Barr virus glycoprotein homologous to herpes simplex virus gB. AB - The Epstein-Barr virus DNA open reading frame BALF4 (R. Baer, A.T. Bankier, M.D. Biggin, P.L. Deininger, P.J. Farrell, T.J. Gibson, G. Hatfull, G.S. Hudson, S.C. Stachwell, C. Sequin, P.S. Tuffnell, and B.G. Barrell, Nature [London] 310:207 211, 1984), which by nucleotide sequence comparison could encode a protein similar to herpes simplex virus gB (P.E. Pellett, M.D. Biggin, B. Barrell, and B. Roizman, J. Virol. 56:807-813, 1985), has now been shown to encode a 110 kilodalton glycoprotein. Late infectious cycle RNAs of 3.0 and 1.8 kilobases are transcribed from BALF4. Translation of these RNAs in vitro, transcription and translation of BALF4 in vitro, or metabolic labeling of cells in the presence of tunicamycin and immunoprecipitation with BALF4-specific sera results in identification of a 93-kilodalton precursor to gp110. Since N-glycosidase F only reduces the size of gp110 to 105 kilodaltons, gp110 probably has both N- and O linked glycosylation, gp110 is an abundant glycoprotein in Epstein-Barr virus infected cells. In infected lymphocytes and in 3T3 cells, in which the gene is expressed from a recombinant expression vector, most of the protein is cytoplasmic and perinuclear. In contrast to gB, gp110 was not detected in the infected-cell plasma membrane. In cells replicating Epstein-Barr virus, gp110 localized to the inner and outer nuclear membrane lamellae and to endoplasmic reticulum structures which sometimes contained enveloped virus. gp110 may play an important role in modifying infected intracellular membranes. PMID- 3027380 TI - Expression of polyomavirus virion proteins by a vaccinia virus vector: association of VP1 and VP2 with the nuclear framework. AB - The polyomavirus proteins VP1, VP2, and VP3 move from their cytoplasmic site of synthesis into the nucleus, where virus assembly occurs. To identify cellular or viral components which might control this process, we determined the distribution of VP1, VP2, and VP3 in a soluble fraction, a cytoplasmic cytoskeleton fraction, and a nuclear framework fraction of infected cells. All three proteins were detected in a detergent-extractable form immediately after their synthesis in polyomavirus-infected cells. Approximately 50, 25, and 40% of pulse-labeled VP1, VP2, and VP3, respectively, associated with the skeletal framework of the nucleus within 10 min after their synthesis. The remaining portion of each labeled protein failed to accumulate on the nuclear framework during a 40-min chase and was degraded. When expressed separately by recombinant vaccinia viruses, VP1 and VP2, but not VP3, accumulated on the nuclear framework. This association was not dependent on other polyomavirus proteins or viral DNA. The amount of total VP1 and VP2 which was bound to the nuclear framework approximated 45 and 20%, respectively. Indirect immunofluorescence demonstrated an exclusive nuclear localization of VP1 in situ. In coinfection experiments, a greater percentage of total VP2 and VP3 was bound to the nuclear framework of cells which cosynthesized VP1. These results indicate that although VP1 and VP2 can bind independently to the insoluble nuclear framework, the association of VP3 with this nuclear structure is promoted by the presence of VP1. PMID- 3027381 TI - Immediate-early genes of murine cytomegalovirus: location, transcripts, and translation products. AB - Cloned genomic fragments from the region (0.769 to 0.818 map units) coding for immediate-early (IE) transcripts of murine cytomegalovirus (MCMV) were used to analyze the physical organization of this region, the direction of transcription, and the proteins synthesized in vitro. Three IE transcription units could be identified. From IE coding region 1 (ie1; 0.781 to 0.796 map units) a dominant 2.75-kilobase (kb) RNA was transcribed from right to left on the prototype arrangement of the MCMV genome which directed the synthesis of an 89,000 molecular-weight polypeptide (89K polypeptide), the major IE protein. This phosphoprotein (pp89) has been shown to be active in the regulation of transcription. Upstream of ie1 and separated by the MCMV enhancer sequence was a second IE coding region, ie2, which was mapped at 0.803 to 0.817 map units. From ie2 a 1.75-kb RNA of moderate abundance was transcribed in the direction opposite to that of the ie1 RNA. After hybrid selection of the ie2 transcript, a 43,000 molecular-weight translation product was detected. A third coding region, ie3, was located directly downstream of ie1 (0.773 to 0.781 map units). The series of RNAs with low abundance, terminating in ie3, probably used the ie1 transcription start site and ranged from 1.0 to 5.1 kb in size. The 5.1-kb RNA apparently represents the nonspliced transcript from both coding regions ie1 and ie3. A 15K polypeptide was translated in vitro from RNA that was hybrid selected by ie3 sequences. Immunoprecipitation with monoclonal antibody revealed that 31K to 67K polypeptides were related to pp89. Some of these proteins were translated from RNAs that were smaller than 2.75 kb. Polypeptides related to pp89 were also synthesized in vivo. Because polypeptides unrelated to pp89 that were translated from RNA that was selected by ie2 and ie3 sequences were not immunoprecipitated by murine antisera, we assumed that the amount of these proteins synthesized in vivo during infection was probably very low. PMID- 3027382 TI - Organization and nucleotide sequence of a densovirus genome imply a host dependent evolution of the parvoviruses. AB - The genome structure of a densovirus from a silkworm was determined by sequencing more than 85% of the complete genome DNA. This is the first report of the genome organization of an insect parvovirus deduced from the DNA sequence. In the viral genome, two large open reading frames designated 1 and 2 and one smaller open reading frame designated 3 were identified. The first two open reading frames shared the same strand, while the third was found in the complementary sequence. Computer analysis suggested that open reading frame 2 may encode all four structural proteins. The genome organization and a part of the nucleotide sequence were conserved among the insect densovirus, rodent parvoviruses, and a human dependovirus. These viruses may have diverged from a common ancestor. PMID- 3027383 TI - Characterization and role in morphogenesis of a new subviral particle (55S) isolated from poliovirus-infected cells. AB - A previously undetected subviral particle, designated the 55S particle because of its position in sucrose density gradients, has been found in cytoplasmic extracts of poliovirus-infected cells. It contains no RNA, is composed of equimolar amounts of the structural polypeptides P1AB, P1C, and P1D, and is stable in vitro under a variety of conditions: presence or absence of EDTA, dilution in low- or high-ionic-strength buffers, suspension in buffers up to pH 10, incubation at 37 degrees C, and centrifugation to equilibrium in CsCl gradients (where it bands at a density of 1.285 g/cm3). Conventional pulse-chase experiments show that 55S particles are the products of the assembly of 14S subunits and the precursors of virions. These data led to the formulation of a model of poliovirus morphogenesis in which the conversion of capsomers into 73S empty capsids does not occur directly, but through the formation of an intermediate structure, the 55S particle. PMID- 3027384 TI - Genetic analysis of myeloproliferative leukemia virus, a novel acute leukemogenic replication-defective retrovirus. AB - The myeloproliferative leukemia virus (MPLV) is a new acute leukemogenic, nonsarcomatogenic retroviral complex that is generated during the in vivo passage of a molecularly cloned Friend ecotropic helper virus. Examination of viral RNA expression in MPLV-producing cells revealed the presence of two distinct molecular species that hybridized with a long terminal repeat or an ecotropic env specific probe but not with a xenotropic mink cell focus-forming virus env specific probe derived from a spleen focus-forming virus: an 8.2-kilobase species corresponding to a full-length Friend murine leukemia virus (F-MuLV) and a deleted species with a genomic size of 7.4 kilobases. This deleted virus was biologically cloned by limiting dilutions and single cell cloning in Mus dunni fibroblasts. Three nonproducer clones with normal morphologies and containing one single integrated copy of the deleted virus were superinfected with F-MuLV, Moloney murine leukemia virus, Gross murine leukemia virus, mink cell focus forming virus (HIX), or the amphotropic 1504 murine leukemia virus. All pseudotypes caused macroscopic and microscopic abnormalities in mice that were similar to those seen in the parental stock. A comparison of the physical maps of F-MuLV and MPLV, which was deduced from the restriction enzyme digests of unintegrated proviral DNAs, indicated that the MPLV-defective genome (i) is probably derived from F-MuLV, (ii) has conserved the F-MuLV gag and pol regions, and (iii) is deleted and rearranged in the env region in a manner that is clearly distinct from that of Friend or Rauscher spleen focus-forming viruses. PMID- 3027385 TI - Structure and function of bicistronic RNA encoding the phosphoprotein and matrix protein of measles virus. AB - Two independent full-length replicas of a bicistronic RNA species containing the complete P and M genes of measles virus arranged in tandem were isolated from an expressible cDNA library. Sequences at the 5' and 3' termini suggested that the bicistronic RNA was initiated and terminated at precisely the same locations as the monocistronic mRNAs of the P and M genes, respectively. The P and M cistrons were fused together via an intergenic region which was exactly colinear with and complementary to the intergenic region of the genomic RNA. This RNA species was polyadenylated at the normal polyadenylation site at the 3' terminus of the M cistron, but not in the intergenic region. By DNA-mediated gene transfer, these cDNA clones were expressed into bicistronic RNA containing both P and M sequences in primate cells. RNA thus generated did not undergo nucleolytic processing but was translated into high levels of a 70,000-Mr protein immunoprecipitated by monoclonal antiserum against the measles virus P protein. M protein was not produced in the same cells even though the M cistron could direct M protein synthesis in vitro once excised from the upstream P cistron. These results suggested that bicistronic RNA could direct protein synthesis from the first but not the second cistron and might contribute at least in part to expression of viral genes in vivo. PMID- 3027386 TI - Laboratory production in vivo of infectious human papillomavirus type 11. AB - Human papillomaviruses (HPV) induce among patients natural lesions which produce small amounts of virus. Infection of human cell cultures does not lead to the multiplication of virus, which also does not replicate in experimental animals. We have developed a unique system for the laboratory production of HPV type 11 (HPV-11). Fragments of human neonatal foreskin were infected with an extract of naturally occurring human vulvar condylomata and grafted beneath the renal capsule of athymic mice. Later (3 to 5 months), condylomatous cysts developed from those grafts. Nuclei of koilocytotic cells contained large amounts of capsid antigen and intranuclear virions. The experimentally induced condylomata were homogenized, and the virions were extracted and used to infect another generation of human foreskin grafts in athymic mice. The HPV-11 DNA content and infectivity of the natural and experimental condylomata were similar. Extracts of experimental condylomata were subjected to differential ultracentrifugation and sedimentation in CsCl density gradients. A single, opalescent band was visible at a density of 1.34 g/ml. It contained HPV virions with HPV-11 DNA. This report is the first demonstration of the laboratory production of an HPV. PMID- 3027387 TI - Herpes simplex virus type 1 glycoprotein E is not indispensable for viral infectivity. AB - A mutant of the herpes simplex virus type 1 Angelotti was isolated in which 87% of the coding region of glycoprotein E (gE) was deleted and replaced by a functional neomycin resistance gene of the Tn5 transposon. The mutant was characterized by restriction enzyme analyses and Southern blotting. Western blotting of proteins and immunofluorescence assays revealed that gE was completely absent and that the Fc receptor was not expressed in cells infected with the mutant. The fact that this mutant was viable and that it replicated to a slightly lower titer than did the wild-type virus suggests that the presence of gE is not a prerequisite of viral infectivity in tissue culture. PMID- 3027389 TI - Differential effects of acyclovir and 9-(1,3-dihydroxy-2-propoxymethyl)guanine on herpes simplex virus and Epstein-Barr virus in a dually infected human lymphoblastoid cell line. AB - We investigated the effects of acyclovir and 9-(1,3-dihydroxy-2 propoxymethyl)guanine (DHPG) on a lymphoblastoid cell line dually infected with Epstein-Barr virus and herpes simplex virus (HSV) type 1. The numbers of Epstein Barr virus genomes were reduced during 70 days of treatment with either drug. Both drugs suppressed HSV replication in a dose-related manner. In the continued presence of the drugs, HSV developed resistance, rapidly to acyclovir and much more slowly to 30 microM DHPG. Analysis of HSV glycoprotein C production and viral DNA showed that treatment with 100 microM DHPG eliminated HSV production, curing the cell line of HSV persistent infection. PMID- 3027388 TI - A mutant of herpes simplex virus type 1 exhibits increased stability of immediate early (alpha) mRNAs. AB - vhs1 is a herpes simplex virus type 1 mutant defective in the shutoff of both host and alpha polypeptide synthesis. In cycloheximide reversal experiments, alpha mRNAs were significantly more stable in vhs1-infected cells than in cells infected with wild-type virus, whether assayed by in vitro translation or Northern blotting. PMID- 3027390 TI - Characterization of product RNAs synthesized in vitro by poliovirus RNA polymerase purified by chromatography on hydroxylapatite or poly(U) Sepharose. AB - The size of the product RNA synthesized by the poliovirus RNA polymerase and host factor was significantly affected by the type of column chromatography used to purify the polymerase. Dimer length product RNA was synthesized by the polymerase purified by chromatography on hydroxylapatite. This contrasted with the monomer length product RNA synthesized by the polymerase purified by chromatography on poly(U) Sepharose. The poly(U) Sepharose-purified polymerase was shown to contain oligo(U) that functioned as a primer. The addition of host factor to reactions containing the poly(U) Sepharose-purified polymerase significantly increased the synthesis of monomer length product RNA, in agreement with previous studies. This product RNA, however, did not immunoprecipitate with anti-VPg antibody and thus was not linked to VPg or a VPg-related protein. Thus, it was concluded that the synthesis of monomer length product RNA by the poly(U) Sepharose-purified polymerase and host factor was caused by oligo(U) priming rather than VPg priming. PMID- 3027392 TI - A nonstructural protein of feline panleukopenia virus: expression in Escherichia coli and detection of multiple forms in infected cells. AB - Sequences coding for the nonstructural protein NS1 of the autonomous parvovirus feline panleukopenia virus were expressed in Escherichia coli as fusion proteins. The fusion proteins were specifically bound by antisera from canine parvovirus infected dogs. Antisera against one of the fusion proteins bound to several proteins found only in feline panleukopenia virus-infected feline cells. PMID- 3027391 TI - Mapping of the transcriptional initiation site of the herpes simplex virus type 1 ICP8 gene in infected and transfected cells. AB - The initiation site for transcription of the herpes simplex virus type 1 (HSV-1) gene encoding the major DNA-binding protein. ICP8, was mapped by nuclease S1 analysis of RNA-DNA hybrids. When RNA isolated from cells infected with HSV-1 was used, one major start site of ICP8 gene transcription was mapped at 89 base pairs to the right of the BstEII site at 0.409 map units. In cells transfected with a cloned ICP8 gene, the same major start site was detected either in the presence or absence of the immediate-early (alpha) genes encoding ICP4 or ICP0, which have been shown to stimulate ICP8 gene expression in transfected cells. Both ICP4 and ICP0 stimulated the accumulation of the ICP8 gene transcripts in the transient expression system, and their effects were synergistic. By comparison of the sequence of the putative promoter region of the ICP8 gene with the promoter of the HSV-1 TK gene, a significant similarity was detected between the three transcriptional regulatory signals of the TK gene and the upstream sequences of the ICP8 gene. Analysis of promoters of other delayed-early (beta) genes showed that they all contained regions of significant homology with the distal signals of the upstream sequences of the TK or ICP8 gene. PMID- 3027393 TI - Interferon induction by transfection of Sendai virus C gene cDNA. AB - To elucidate the mechanism of interferon (IFN) induction on virus infection, we constructed two types of plasmids by inserting a part of the cDNA of the Sendai virus into a simian virus 40-derived expression vector (pSV2-0). One, pSV2-PC, contained the P + C gene, which codes for the P and C proteins in overlapping reading frames, and the other, pSV2-C, contained only the C gene. After transfecting the plasmids into mammalian cells, we determined the IFN activity in the culture medium. We found that the level obtained with pSV2-PC was significantly positive but very low, whereas that obtained with pSV2-C was as high as or even higher than that observed in the culture medium after Sendai virus infection. By cleaving pSV2-C between the simian virus 40 promotor and the C gene or by inserting a stop codon within the C gene [pSV2-C(stop)], induction of IFN was greatly diminished. In Northern blot analyses of the transcripts obtained from the cells transfected with the plasmids with cDNA to the P + C gene as a probe, the transcript having the expected size was detected with both pSV2-C and pSV2-C(stop), whereas none was detected with cleaved pSV2-C or pSV2-0. The results indicate that both transcription and translation of the C gene seem to be required for IFN induction after Sendai virus infection. PMID- 3027394 TI - Inhibition of vesicular stomatitis viral mRNA synthesis by interferons. AB - Interferon (IFN) treatment inhibited the replication of vesicular stomatitis virus (VSV) in human GM2767 and mouse JLSV-11 cells. The replication of this virus in either human RD-114 or mouse A402 cells was insensitive to IFN treatment. We analyzed various steps in the VSV life cycle as they occurred under different conditions of IFN treatment to identify the point(s) at which IFN was exerting its inhibitory effect. IFN treatment led to strong inhibition of viral protein synthesis and accumulation of viral RNA in both lines of IFN-sensitive cells. No such effect was observed in the IFN-resistant cells. Using a temperature-sensitive mutant (tsG41) and wild-type VSV that were not undergoing protein synthesis, we determined that the major site of action of IFN against VSV replication in JLSV-11 and GM2767 cells was at the level of primary viral transcription. The accumulation of primary viral transcripts was strongly inhibited in these cells by IFN treatment. This effect was not a consequence of any effect of IFN on virus entry and uncoating. Thus, it appears that IFN exerts a direct effect on the VSV transcriptional process in GM2767 and JLSV-11 cells. PMID- 3027396 TI - Germ line integration of a murine leukemia provirus into a retroviruslike sequence. AB - Nucleotide sequence analysis of the cellular sequences flanking the integrated ecotropic (mouse-infectious) murine leukemia provirus of BALB/c mice indicated that the murine leukemia provirus is integrated in opposing transcriptional orientation within a solo long terminal repeat (LTR) of the VL30 family of endogenous retrovirus-related sequences. To quantify the effect of this integration event on the ability of the ecotropic provirus to be expressed, we constructed recombinant molecules that carried the chloramphenicol acetyltransferase (cat) gene and various viral LTRs and determined the CAT activity induced by these constructs after transfection of NIH 3T3 cells. Our results indicate that the BALB/c ecotropic LTR is about 10-fold more active than the VL30 LTR. The presence of the VL30 LTR did not affect the transcriptional activity of the ecotropic LTR in the context of the integration event. Our results also indicate that the LTRs of the BALB/c provirus are less transcriptionally active than are the proviral LTRs of AKR murine leukemia virus and the Harvey murine sarcoma virus. PMID- 3027397 TI - Abundant synthesis of functional human T-cell leukemia virus type I p40x protein in eucaryotic cells by using a baculovirus expression vector. AB - The human T-cell leukemia virus type I (HTLV-I) p40x protein is a 40-kilodalton polypeptide encoded in the 3'-terminal region of the virus. This protein is responsible for positive transcriptional trans-activation of promoter elements located within the HTLV-I long terminal repeat. We introduced the protein-coding region of HTLV-I p40x into the genome of the baculovirus Autographa californica nuclear polyhedrosis virus. After infection of the insect Spodoptera frugiperda (SF9) cell line, this recombinant strain of baculovirus produced approximately 200 mg of intact p40x protein per 2.5 X 10(8) cells. The protein was biologically active in trans-activation of an HTLV-I long terminal repeat-human beta-globin construct. Biochemical analyses of the protein suggest that the p40x polypeptide underwent posttranslational modification in these eucaryotic SF9 cells. PMID- 3027395 TI - Adenovirus E1A 12S protein induces DNA synthesis and proliferation in primary epithelial cells in both the presence and absence of serum. AB - Infection of primary baby rat kidney (BRK) cells with an adenovirus that carries an E1A 12S cDNA in place of the normal E1A region (adenovirus 5 [Ad5] 12S) resulted in the induction of cellular DNA synthesis and proliferation of the epithelial cells in the population, even in the absence of serum. Increased cellular DNA synthesis was first detectable by 12 h after infection and was maintained at a 10- to 20-fold higher level than in mock-infected cells. By 5 days after infection there was a 10-fold-greater number of 12S virus-infected BRK cells. These infected BRK cells retained many of their normal epithelial cell characteristics and were not transformed. The expression of the E1A 12S protein(s) occurred early after infection. There was no induction of adenoviral gene expression or viral DNA replication in these cells. The early effects of a fully transforming gene product(s) were also examined. The Ad5-simian virus 40 hybrid virus, Ad5.SVR4, in which the early region of simian virus 40 has replaced the E1 region of Ad5, was used to infect BRK cells. The kinetics of expression of the T antigens were similar to those of the 12S polypeptides. Infection with Ad5.SV4 also resulted in the induction of cellular DNA synthesis and cell proliferation at levels similar to those observed with the 12S virus. However, infection with Ad5.SVR4 resulted in cells that had lost some of their epithelial cell characteristics and were fully transformed. Thus, although the early cellular events induced by the two genes were similar, they did not yield the same final cellular phenotype. PMID- 3027398 TI - Linker-insertion nonsense and restriction-site deletion mutations of the gB glycoprotein gene of herpes simplex virus type 1. AB - To study the effects of missense, nonsense, and deletion mutations of the gB glycoprotein gene of herpes simplex virus type 1, a gB-transformed cell line was isolated that, after virus infection, would express sufficient quantities of gB from the cellular chromosome to complement temperature-sensitive gB mutants. The transformed cell line was then used as a permissive cell to transfer two gB mutations from plasmid to viral DNA. One of the mutants, K082, harbored an HpaI linker insertion that introduced one new amino acid and a chain terminator codon within amino acid residue 43. The other mutant contained a 969-base-pair deletion in a part of the gene that includes the membrane-spanning region; a correspondingly shorter gB polypeptide was detected by sodium dodecyl sulfate-gel electrophoresis after immunoprecipitation of infected-cell extracts with four pooled monoclonal antibodies. No polypeptide was observed from K082-infected cells. The shortened gB polypeptide was efficiently processed and secreted into the growth medium. Each of the four monoclonal antibodies precipitated full length gB, and three of the four precipitated the shortened polypeptide. Enveloped virus particles could be purified after infection of nonpermissive cells with either mutant virus. Virus particles appeared to possess normal polypeptide and glycopeptide profiles except for the absence of gB. Therefore, the presence of gB is not essential for viral assembly, including envelopment. Recombinants in virus stocks grown on the gB-transformed cells occurred at frequencies on the order of 10(-7) to 10(-5), compared with a frequency of approximately 10(-2) in mixed infections with the two mutants. PMID- 3027399 TI - Embryonic infection with the endogenous avian leukosis virus Rous-associated virus-0 alters responses to exogenous avian leukosis virus infection. AB - We inoculated susceptible chicken embryos with the endogenous avian leukosis virus Rous-associated virus-0 (RAV-0) on day 6 of incubation. At 1 week after hatching, RAV-0-infected and control chickens were inoculated with either RAV-1 or RAV-2, exogenous viruses belonging to subgroups A and B, respectively. The chickens injected with RAV-0 as embryos remained viremic with exogenous virus longer and either failed to develop type-specific humoral immunity to exogenous virus or developed it later than the control chickens not inoculated with RAV-0. The RAV-0-injected chickens also developed neoplasms at a much higher frequency than did the control chickens. We suggest that the lower immune responses of the RAV-0-injected chickens were due to an immunological tolerance to envelope group specific glycoproteins shared among endogenous and exogenous viruses. PMID- 3027401 TI - Characterization of equine infectious anemia virus long terminal repeat. AB - The long terminal repeats (LTRs) of equine infectious anemia virus (EIAV) were examined with respect to their ability to function as transcriptional promoters in various cellular environments. Nucleotide sequence analyses of the LTRs derived from two unique proviral clones revealed the requisite consensus transcription and processing signals. One of the proviruses possessed a duplication of a 16-base-pair sequence in the CCAAT box region of the LTR which was absent in the other provirus. To assess its functional activity, each LTR was coupled to the bacterial chloramphenicol acetyltransferase gene and transfected onto various cell lines, including matched cultures of EIAV-infected and uninfected cells. The levels of chloramphenicol acetyltransferase activity directed by the EIAV LTRs were between 250 and 900 times greater in EIAV-infected cells compared with their uninfected counterparts. Thus, EIAV expression appears to be activated by a virus-induced trans-activation phenomenon analogous to that recently shown to amplify expression of certain other lentiviruses. PMID- 3027400 TI - Mechanisms of antiviral immunity induced by a vaccinia virus recombinant expressing herpes simplex virus type 1 glycoprotein D: cytotoxic T cells. AB - We used a transfected L cell and a vaccinia vector carrying the herpes simplex virus type 1 (HSV-1) gene coding for glycoprotein D (gD) to characterize HSV specific T-cell responses. Various studies with mice revealed that the vectors could stimulate some HSV-specific T-cell responses. Although the majority of the T cells contributing to the HSV-1 gD-specific proliferative response were of the Lyt-2.1+ phenotype, cytotoxic T cells (Tc), surprisingly, were not induced by these gD vectors. Even though gD appeared to be a target for a class II major histocompatibility complex (MHC)-restricted killer cell, neither gD vector was capable of forming a target cell complex which could be recognized by class I MHC restricted HSV-specific Tc. Further investigation of the gD-specific responses revealed the presence of potent suppressor cells and factors capable of inhibiting HSV-specific Tc induction in in vitro assays. One interpretation of these data is that class I MHC-restricted HSV- and gD-specific Tc do not develop during HSV infection because of active suppression. PMID- 3027402 TI - Simian virus 40 associates with nuclear superstructures at early times of infection. AB - The association of infecting simian virus 40 with insoluble nuclear structures was assayed by disrupting infected nuclei and assaying insoluble fractions for virus. Three methods were used which lyse nuclei but maintain the insolubility of residual nuclear structures: sonication, high-salt-Triton-EDTA extraction, and low-salt-lithium diiodosalicylate extraction. After each type of nuclear extraction, infecting simian virus 40 remained associated with the residual nuclear structures. This association depended strictly on natural viral infections and on the use of buffers containing moderate amounts of salt and Mg2+ for the isolation of infected nuclei. These viral interactions exhibited behavior similar to host cell DNA interactions studied by analogous assays. Both viral DNA and coat proteins were found associated with the host cell nuclear superstructure. We concluding that at early times after infection the viral templates mimic the state of the host cell chromatin by attaching to the cellular nuclear matrix. PMID- 3027403 TI - Derivation and characterization of POJ cells, transformed human fetal glial cells that retain their permissivity for JC virus. AB - The study of the medically important polyomavirus JC virus is limited to only a few laboratories, primarily because the permissive cell system most often used, primary human fetal glial cells, is difficult to obtain and propagate. We have introduced mutations at the origin of DNA replication of JC virus and transformed glial cells with the replication-defective genomes. Although normal glial cell cultures rapidly lose their permissivity for the virus after subculture, the transformed cells (designated POJ) had a greatly expanded life span and remained permissive for JC virus even after 30 passages in vitro. POJ cells constitutively express a functional T protein that complements the replication defect of lethal early-region mutations in JC virus. We expect that these cells will greatly facilitate the study of this human virus. PMID- 3027404 TI - Synergistic activation of cells by Epstein-Barr virus and B-cell growth factor. AB - Infection with Epstein-Barr virus (EBV) is initiated by virus binding to the C3dg C3d receptor CR2. Several workers have implicated this receptor in the control of B-cell activation by examining the effects of antibodies to CR2 and isolated C3d on B-cell proliferation and differentiation. We report here on the activating effects of irradiated EBV, which retains its capacity to bind to CR2 but loses its ability to function as a T-independent B-cell activator. EBV synergized with B-cell growth factor in the induction of uptake of tritiated thymidine by T cell depleted leukocytes from seronegative donors but did not induce secretion of immunoglobulin. Synergism could be inhibited with an anti-viral antibody that inhibited binding of EBV to CR2. No similar synergism was found between EBV and recombinant interleukin 2, interleukin 1 alpha, or gamma interferon or with the lipid A fraction of bacterial lipopolysaccharide. EBV may thus initiate B-cell activation as it binds to CR2. Infectious virus may, under normal circumstances, induce the cell to make those growth factors necessary to support B-cell proliferation; the difficulty of transforming cells with transfected EBV DNA may in part reflect the absence of an activation event provided by intact virus as it attaches to CR2. The synergism of EBV and B-cell growth factor more clearly distinguishes the effects of B-cell growth factor from those of interleukin 1 and interleukin 2 in other models of B-cell activation. Thus, this may be a useful model for further delineation of unique effects of B-cell growth factor on B-cell function. PMID- 3027405 TI - Adenovirus early region 1A modulation of interferon antiviral activity. AB - Human adenovirus type 5 (Ad5) is a DNA virus which replicates as efficiently in human A549 cells treated with human interferon-alpha 2 (IFN) as in untreated cells. Vesicular stomatitis virus (VSV), on the other hand, is a negative-strand RNA virus which is very sensitive to the effects of IFN treatment in A549 cells. The IFN-mediated inhibition of VSV replication was not observed in cells coinfected with Ad5. Abrogation of IFN-mediated antiviral activity was maximal when Ad5 infection preceded VSV infection by at least 36 h, but did not require adenovirus DNA synthesis for manifestation. Coinfection experiments with VSV and deletion variants of adenovirus demonstrated that neither virus-associated RNA synthesis nor expression of adenovirus early regions E1B, E2A, E3, or E4 are required for abrogation of IFN-mediated inhibition of VSV replication. However, expression of early region E1A was essential, suggesting that E1A products can modulate, either directly or indirectly, IFN activity in adenovirus-infected cells. PMID- 3027406 TI - Genome location and identification of functions defective in the Bartha vaccine strain of pseudorabies virus. AB - We have shown previously (Lomniczi et al., J. Virol. 52:198-205, 1984) that the Bartha vaccine strain of pseudorabies virus has a deletion in the short unique (Us) region of its genome--a deletion that is related to the absence of virus virulence. This strain is, however, also defective in other genes involved in virulence. We show here that virulence can be restored by marker rescue of the Bartha strain to which an intact Us has been restored (but not to the parental Bartha strain) by sequences derived from approximate map units 0.460 and 0.505 of the wild-type virus genome. No difference in the ability to grow in cell culture was observed between parental Bartha, Bartha 43/25a (Bartha to which an intact Us has been restored), or the doubly rescued Bartha strains. However, only the doubly rescued Bartha strain was virulent for both chickens and pigs and replicated to high titers when inoculated directly into the brains of chickens. The sequences that could restore virulence to the Bartha 43/25a strain encode four genes, all of which are involved in processes leading to the assembly of nucleocapsids. Since these sequences rescue virulence, it appears that a function that plays a role in nucleocapsid assembly is defective in the Bartha strain and that this defect contributes to the lack of virulence of this virus. PMID- 3027408 TI - Deletion mutants in the gene encoding the herpes simplex virus type 1 immediate early protein ICP0 exhibit impaired growth in cell culture. AB - We report the construction and characterization of deletion mutants in the herpes simplex virus type 1 gene encoding the immediate-early protein ICP0. In the event that ICP0 proved to play an essential role in virus replication, ICP0-transformed Vero cells were generated to serve as permissive hosts for such mutants. Two mutants, dlX0.7 and dlX3.1, were isolated in these cells by a marker rescue transfer procedure involving the rescue of an ICP4 deletion mutant and the simultaneous insertion of a linked deletion in the ICP0 gene. Mutant dlX0.7 contained a 700-base-pair deletion in both copies of ICP0. The deletion lay entirely within the transcript specified by the gene. dlX0.7 induced the synthesis of an ICP0-specific mRNA that was approximately 0.7 kilobases smaller than the corresponding mRNA specified by wild-type virus. The 3.1-kilobase deletion in both copies of the ICP0 gene in mutant dlX3.1 removed the majority of the transcriptional-regulatory signals and coding sequences, retaining only sequences at the 3' end of the gene. As expected, no ICP0-specific mRNA was detected in dlX3.1-infected Nero cells (G418-resistant Vero cells). Both mutants grew in all cells tested, although their burst sizes were 10- to 100-fold lower than that of wild-type virus. Although the plaque sizes of dlX0.7 and dlX3.1 were equally small on Nero and ICP0-transformed cells, the plating efficiency of the mutants was 15- to 50-fold greater on ICP0-transformed cells than on Nero cells. The mutants exhibited modest interference with the growth of wild-type virus in mixed infections, an effect that was abolished by UV irradiation of the mutants, implying that interference required viral gene expression. Polypeptide profiles generated by the mutants in Nero cells were qualitatively similar to that of wild type virus. Quantitatively, only slight reductions in the levels of certain late viral polypeptides were observed, a phenomenon also borne out by analysis of viral glycoproteins. Both mutants induced the synthesis of significant, although reduced, levels of viral DNA relative to wild-type virus. Taken together, the results demonstrate that ICP0 is not essential for productive infection in cell culture but that this protein plays a significant role in viral growth, as indicated by the impaired abilities of the mutants to replicate. PMID- 3027407 TI - Host factor-induced template modification during synthesis of poliovirus RNA in vitro. AB - Poliovirus RNA polymerase, 3Dpol, transcribes poliovirus RNA in vitro in the presence of a host factor (HF) preparation from uninfected HeLa cells to yield heterogeneous-size product RNAs. The products include some molecules larger than the template that represent self-primed elongations of the template from a 3' terminal hairpin. We showed that transcription proceeded through the formation of a modified RNA intermediate that was generated by nucleolytic cleavage of the template by HF in the absence of nucleoside triphosphates. Cleavage resulted in the loss of the original poly(A) 3' end and the generation of new, heterogeneous 3' ends that formed self-priming structures that could then be elongated by 3Dpol or reverse transcriptase. The two stages of the reaction, (i) cleavage to yield self-priming templates and (ii) subsequent chain elongation to yield heterogeneous-size products up to nearly dimer length, could be separated. RNAs whose original 3' ends were chemically oxidized so as to prevent chain elongation showed no reduction in template activity after preincubation with HF. We conclude that an HF preparation that contains a low level of nuclease activity is sufficient to activate RNA templates for transcription by 3Dpol to generate up to apparent dimer-length products. This reaction likely has little relevance to the mechanism of poliovirus RNA replication in vivo. It is likely that numerous other factors or activities, in addition to 3Dpol, would also result in transcription of poliovirus RNA in vitro. PMID- 3027409 TI - Resolution of a polyomavirus-mouse hybrid replicon: release of genomic viral DNA. AB - RmI is a circular chimera containing 1.03 copies of polyomavirus DNA and 1,628 base pairs of mouse DNA, joined through direct and inverted repeat sequences. It is excised from the chromosome of a transformed cell via a site-specific recombination event that is dependent on the activation of the viral gene coding for large T antigen. RmI is shown here to be highly infectious for normal mouse cells. This infectivity reflects the ability of RmI to effectively yield unit length viral DNA via intramolecular recombination. The effectiveness with which infectious viral DNA is produced from RmI is consistent with the idea that the underlying recombination event is site specific, rather than homologous or illegitimate. PMID- 3027410 TI - Resolution of a polyomavirus-mouse hybrid replicon: viral function required for recombination. AB - RmI, a circular chimera made of the polyomavirus (Py) genome with an insertion of mouse DNA (Ins), effectively undergoes intramolecular recombination in normal mouse cells, as indicated by the conversion of cloned RmI (RmIc) into unit-length Py DNA in transfected cultures. To follow the fate of the cellular component of RmI after recombination, the origin of simian virus 40 (SV40) DNA was inserted into the Ins region of RmIc, generating a new molecular species designated SV RmIc. The recombination of SV-RmIc in simian cells synthesizing SV40 large T antigen gave rise to a molecule containing the SV40 origin, the reciprocal of unit-length Py DNA. However, SV-RmIc failed to yield unit-length Py DNA in murine cells unless Py large T antigen was provided in trans. In murine cells synthesizing SV40 large T antigen, the only detectable product from SV-RmIc contained only Py sequences, but was heterogeneous in size. These results and others also reported here strongly suggest that Py large T antigen plays a direct role in the resolution of RmI in murine cells. PMID- 3027411 TI - Analysis of nonpermissivity in mouse cells overexpressing simian virus 40 T antigen. AB - To analyze the nature of the nonpermissivity of mouse cells for simian virus 40 (SV40) DNA replication, we isolated mouse cells producing SV40 T antigen (Tag) at levels equal to or greater than that found in COS1 cells. These mouse cells were nonpermissive for the replication of exogenously added SV40 DNA, although purified Tag isolated from these cells was able to support SV40 DNA replication in vitro. Furthermore, when mouse cells expressing Tag were fused to monkey cells, SV40 DNA replication was observed. These results indicate that the mere production of large quantities of wild-type SV40 Tag does not overcome the block of nonpermissivity in mouse cells and that cellular factors must play a critical role. PMID- 3027412 TI - Binding of the herpes simplex virus immediate-early gene product ICP4 to its own transcription start site. AB - A gel electrophoresis DNA-binding assay was used to detect proteins from herpes simplex virus type 1-infected and uninfected cells that specifically bind the upstream region of immediate-early (IE) gene 3. The assay is based on the altered electrophoretic mobility of DNA-protein complexes relative to that of free DNA in native gels. A series of end-labeled overlapping DNA fragments spanning a region from -272 to +27 (relative to the 5' terminus of the IE gene 3 mRNA) were used as probes. Two complexes were identified (referred to as A and B) which were driven by different protein factors. Formation of the A complex required infected-cell proteins extracted at any time from 2 to 16 h postinfection; a 0.5 to 1 M NaCl extract of infected cells, and a DNA probe that contained the sequences from -4 to +27 (relative to the 5' terminus of IE gene 3 mRNA). The protein that drove the formation of the A complex is not related to transcription factors TFIIIA or Sp1 or their cognate binding domains since neither the 5S RNA gene nor the GC box of simian virus 40 could compete for proteins that induced formation of the A complex. Through the use of monoclonal antibodies, the complex was shown to contain the IE gene 3 product, ICP4. A more detailed localization of the DNA binding site in vitro by using chemical footprinting revealed that binding occurs over the sequence from -10 to +3 relative to the mRNA terminus. The binding of ICP4 to its own transcription start site may explain the repression of IE gene transcription which attends the onset of early (beta) gene expression and suggests an autoregulatory mechanism for gene control in herpes simplex virus type 1. The B complex was readily detected in uninfected cells (of a number of different cell lines), as well as in infected cells, with a probe containing the IE consensus sequence TAATGARATTC (where R is a purine) and two nested copies of the Sp1 binding motif GGGCGG; however, complexes were also detected with probes that lack the IE consensus sequence but contain Sp1 sites. These data suggest that the B complex contains the promoter-specific factor Sp1, and competition experiments with the clustered Sp1 binding domains from simian virus 40 confirmed this idea. PMID- 3027414 TI - Expression of mink cell focus-forming murine leukemia virus-related transcripts in AKR mice. AB - We used a synthetic 16-base-pair mink cell focus-forming (MCF) env-specific oligomer as radiolabeled probe to study MCF murine leukemia virus (MuLV)-related transcripts in brain, kidney, liver, spleen, and thymus tissues of AKR mice ranging from 5 weeks to 6 months (mo) of age. Tissue-specific expression of poly (A) + RNAs was seen: 6.0-kilobase (kb) transcripts were detected in the liver and kidney; 7.2- and 1.8-kb RNA species were present in the thymus. In addition, all the tissues tested contained 3.0-kb messages. The transcription of these MCF related mRNAs was independent of the presence of ecotropic and xenotropic MuLVs. In general, expression of the MCF env-related transcripts appeared to peak at 2 mo of age; these messages were barely detectable in brain, kidney, liver, and spleen tissues after 2 mo and in thymus tissue after 4 mo of age. All of the subgenomic MCF env-related mRNAs (6.0, 7.2, 1.8, and 3.0 kb) appeared to contain the 190-base-pair cellular DNA insert, characteristic of the long terminal repeats associated with endogenous MCF env-related proviruses (A. S. Khan and M. A. Martin, Proc. Natl. Acad. Sci. USA 80:2699-2703, 1983). No genomic-size (8.4 kb) transcripts corresponding to endogenous MCF-related proviruses were detected. An 8.4-kb MCF env-related mRNA was first seen at 3 mo of age, exclusively in thymus tissue. This species most likely represents the first appearance of a recombinant MCF-related MuLV genome. The transcripts which were detected in thymus tissue might be involved in the generation of leukemogenic MCF viruses. PMID- 3027415 TI - A temperature-sensitive mutant of the myeloproliferative sarcoma virus, altered by a point mutation in the mos oncogene, has been modified as a selectable retroviral vector. AB - The myeloproliferative sarcoma virus (MPSV) is a mos-oncogenic retrovirus which induces an acute myeloproliferative disease in adult mice. The isolation and molecular cloning of two mutants of MPSV temperature sensitive (ts) for mos transformation (Kollek et al., J. Virol. 50:717-724, 1984) have been described previously. In this report, we describe the biological activity of these clones, the molecular basis of the ts lesion of one clone, and the construction of a selectable vector based on the MPSV ts genome. Both molecular clones, ts159 and ts124, proved to have retained the ts phenotype, the former being tighter for the induction and maintenance of the transformed phenotype. A single transition (G--- A) at position 1888 in the mos coding region, resulting in the change of Gly to Arg at position 307, was responsible for the ts phenotype of clone ts159. Substitution of sequences carrying this mutation with the corresponding sequences of the wild-type virus generated a virus that was ts for transformation. Insertion of the dominant selectable marker gene for geneticin resistance (neor) into ts159 did not disrupt mos expression or its ts phenotype. neor-ts159 facilitates the study of mos action by allowing the selection of infected cells at the nonpermissive temperature before mos transformation has been induced. Furthermore, infected cells which show no obvious phenotype alteration due to mos expression can be identified by their Neor phenotype. PMID- 3027413 TI - Posttranslational processing of an Epstein-Barr virus-encoded membrane protein expressed in cells transformed by Epstein-Barr virus. AB - The BamHI Nhet fragment of the B958 strain of Epstein-Barr virus (EBV) encodes a membrane protein (BNLF-1) that is present in cells transformed by EBV. We made a hybrid protein in which a polypeptide sequence from the carboxyl-terminal part of BNLF-1 is fused to Escherichia coli beta-galactosidase. This hybrid protein was used to immunize rabbits, and the resulting antiserum was purified by immunoaffinity chromatography. The antiserum was able to immunoprecipitate BNLF-1 from cell lysates. We found that BNLF-1 is phosphorylated at serines in EBV genome-positive B-cell lines. Pulse-chase analyses with [35S]methionine indicated that BNLF-1 is turned over in lymphoblasts with a half-life of approximately 5 h. Protein immunoblots of EBV genome-positive B-cell lines revealed both a 62,000 molecular-mass band corresponding to BNLF-1 and a myriad of lower-molecular-mass bands. We postulate that these lower-molecular-mass bands are degradation products resulting from the turnover of BNLF-1 in cells. The BNLF-1 gene was expressed in COS cells, and the protein was both phosphorylated and turned over in these cells. PMID- 3027416 TI - Macrophage-resistant murine simian virus 40 tumors express a retroviral type specific gp70. AB - Prototype macrophage-resistant and -sensitive cells were subcloned. Among several subclones of the resistant line, one subclone showed partial reversion to a sensitive phenotype. Analysis with monoclonal antibodies specific for different serotypes of endogenous murine leukemia virus revealed that expression of only one such gp70 (gp70a) correlated with the macrophage-resistant phenotype. PMID- 3027417 TI - env-encoded residues are not required for transformation by p48v-myb. AB - The v-myb oncogene of avian myeloblastosis virus induces acute myeloblastic leukemia in chickens and transforms avian myeloid cells in vitro. The protein product of this oncogene, p48v-myb, is partially encoded by the retroviral gag and env genes. We demonstrated that the env-encoded carboxyl terminus of p48v-myb is not required for transformation. Our results showed, in addition, that a coding region of c-myb which is not essential for transformation was transduced by avian myeloblastosis virus. PMID- 3027419 TI - Dimers and complexes with p53 are the prevalent oligomeric forms of a transforming nonkaryophilic T antigen of simian virus 40. AB - The oligomers formed by a mutant nonkaryophilic large T antigen of simian virus 40, which lacks residues 110 through 152 of normal large T antigen and transforms only established cells (L. Fischer-Fantuzzi and C. Vesco, Proc. Natl. Acad. Sci. USA 82:1891-1895, 1985), were found to consist predominantly of dimers. Anti-p53 antibodies precipitated 14 to 16S complexes containing the mutant nonkaryophilic large T antigen and p53 from extracts of transformed cells, and anti-p53 indirect immunofluorescence stained these cells in the cytoplasm. PMID- 3027418 TI - Role of the agnoprotein in regulation of simian virus 40 replication and maturation pathways. AB - Analysis of two agnogene mutants, dl2304 deleted over the entire agnogene and in2379 carrying a 2-base insert, indicated that the mutant phenotype of small plaque formation must be the result of a defect late in the maturation pathway. Both mutants were removed from the pool of molecules available for replication with wild-type kinetics. Whereas dl2304 was somewhat reduced in its rate of progression from chromatin to previrions-virions, in2379, which produced even smaller plaques than dl2304 did, progressed with wild-type kinetics. Therefore, the agnoprotein was not required for progression from chromatin to previrions. PMID- 3027420 TI - Computerized tomography demonstration of an intrarenal teratoma. AB - The computerized tomographic appearance of an intrarenal teratoma is described. The differential diagnoses, especially from Wilms tumor, are discussed. PMID- 3027421 TI - gamma-Carboxyglutamic acid, a component in human pediatric bladder stones containing calcium salts. AB - The amino acid gamma-carboxyglutamic acid (Gla) has been previously detected in the vitamin K-requiring blood clotting factors, proteins of calcified vertebrate tissue, renal tissue, plasma protein C, ectopic calcifications, and calcium containing renal calculi. This paper reports the presence of Gla in the EDTA soluble, non-dialyzable material recovered from human pediatric bladder stones containing calcium salts. In bladder stones composed of calcium oxalate, uric acid and ammonium acid urate, 73 Gla residues per 1,000 amino acid residues were detected. Bladder stones composed of calcium oxalate, uric acid, ammonium acid urate, and hydroxyapatite contained 48 Gla residues per 1,000 amino acid residues present. No Gla was detected in the predominantly magnesium ammonium phosphate (struvite) bladder stones. These results with human bladder stones from children under 10 years of age are consistent with the observations from adult patients in which Gla was detected in the calcium-containing renal calculi but not in the non calcium-containing renal calculi. The present study adds to the growing body of information concerning the possible role of Gla in normal and abnormal calcium metabolism. PMID- 3027422 TI - Postservice mortality among Vietnam veterans. The Centers for Disease Control Vietnam Experience Study. AB - The postservice mortality (through December 1983) of a cohort of 9324 US Army veterans who served in Vietnam was compared with that of 8989 Vietnam-era Army veterans who served in Korea, Germany, or the United States. Over the entire follow-up period, total mortality in Vietnam veterans was 17% higher than for other veterans. The excess mortality occurred mainly in the first five years after discharge from active duty (rate ratio, 1.45; 95% confidence interval, 1.08 to 1.96) and involved motor vehicle accidents, suicide, homicide, and accidental poisonings. Thereafter, mortality among Vietnam veterans was similar to that of other Vietnam-era veterans, except for drug-related deaths, which continued to be elevated. An unexpected finding was a deficit in deaths from diseases of the circulatory system among Vietnam veterans. The excess in postservice mortality due to external causes among Vietnam veterans is similar to that found among men returning from combat areas after World War II and the Korean War. PMID- 3027424 TI - Prevention of HIV infection. PMID- 3027423 TI - Combination oral contraceptive use and the risk of endometrial cancer. The Cancer and Steroid Hormone Study of the Centers for Disease Control and the National Institute of Child Health and Human Development. AB - To examine the relationship between endometrial cancer and use of specific oral contraceptive (OC) formulations, we used data from the Cancer and Steroid Hormone Study, a multicenter, population-based, case-control study. Cases were 433 women aged 20 to 54 years with histologically confirmed epithelial endometrial cancer ascertained through six population-based cancer registries. Controls were 3191 women selected at random from the populations of these areas. Women who had used combination OCs for at least 12 months had an age-adjusted risk of developing endometrial cancer of 0.6 relative to those women who had never used OCs (95% confidence interval, 0.3 to 0.9). This protective effect persisted for at least 15 years after the cessation of OC use. Examination of the eight most frequently used OC formulations revealed little difference in the age-adjusted risks, which ranged from 0.2 to 0.7 for women who had ever used a formulation compared with women who had never used OCs. Use of OCs for 12 months or longer conferred protection against all three major histologic subtypes of endometrial cancer. PMID- 3027425 TI - Need for caution in interpretation of Western Blot tests for HIV. PMID- 3027427 TI - Beta-adrenergic receptor-adenylate cyclase system in the myocardium of spontaneously hypertensive rats and changes in the components of this system during aging. PMID- 3027426 TI - Herpes simplex related antigen in cases of carcinoma of the oral cavity. PMID- 3027428 TI - [The control of hypertension with dibutyryl cyclic AMP under general anesthesia]. PMID- 3027429 TI - [An autopsy case of meningeal carcinomatosis with vestibulocochlear nerve disturbance as the first manifestation]. AB - A 54-year-old man initially complained of frontal headache, right ear pain and tinnitus in May, 1985. This was followed by right facial palsy and hearing loss, and he was admitted to our hospital. Physical findings revealed right trigeminal nerve disturbance, left facial nerve palsy and bulbar palsy. The spinal fluid showed pleocytosis, increased protein, decreased glucose, markedly increased carcinoembryonic antigen and adenocarcinoma cells. Gastric carcinoma was revealed by an upper GI series. He was treated with chemotherapy. However, he die in August, 1985. Nodular metastases were discovered at the right internal acoustic meatus and other areas. Microscopically, signet-ring cell carcinoma had diffusely infiltrated at the subarachnoid space. PMID- 3027430 TI - [A case of secondary linitis plastica of the colon developing 14 years after gastrectomy in advanced carcinoma]. AB - A 59-year-old woman was admitted to our hospital complaining of weight loss. Fourteen years earlier, she had undergone gastrectomy for gastric carcinoma of the Borrmann III type on the lesser curvature of the body. Biopsy specimens of the lesion revealed poorly differentiated adenocarcinoma. On the fifth hospital day of the admission under study, she complained of vaginal bleeding, and a diagnosis of endometrial carcinoma was made by a gynecologist. Transverse colectomy and total hysterectomy were done. The lesion was localized only in the transverse colon, was histologically poorly differentiated adenocarcinoma similar to the specimen of gastric carcinoma resected 14 years earlier and involved mainly the serosa. Therefore, secondary linitis plastica of the transverse colon was diagnosed. PMID- 3027432 TI - Prophylactic efficacy of immune serum globulin against hepatitis A. AB - A remarkable decline in the morbidity of hepatitis A was observed in the members of the Japan Overseas Cooperation Volunteers (JOCV) following inoculation of these volunteer workers with human immune serum globulin (ISG) once every 4-6 months, a program which was started in 1981 on a voluntary basis. The morbidity rate of hepatitis A declined from 3.5% in 1980 to 0.6% in 1983 and 1984. A significant difference in the morbidity of hepatitis A was observed among those who were not. In addition to precautions about drinking water and fresh or uncooked foods, inoculation with ISG for the prophylaxis against hepatitis A is recommended when non-immune subjects from a developed country travel to or stay in developing countries. PMID- 3027431 TI - Demonstration of herpes simplex 1 virus antigen mouse brain after fixation. AB - The effects of several fixatives and fixation periods on recovery of viral antigen by indirect immunofluorescence (IF) and indirect immunoperoxidase (IP) were examined in mouse brains infected with type-1 herpes simplex virus. In 10% formalin, the sensitivity of antigen detection by IF decreased after one month fixation and viral antigen was barely detectable after fixation for three months. But in 10% buffered formalin antigenicity was not lost after six months. Similar effects were obtained by IP; however, specimens fixed in 10% formalin for three and six months showed positive reactions. Viral antigen was detectable without enzyme digestion in paraffin materials fixed in Bouin's, Carnoy's and Dubosq Brazil solutions, but when treated with proteolytic enzyme for three hours, antigen could not be detected by either IF or IP. Ten per cent buffered formalin, Bouin's or Dubosq-Brazil solutions are recommended as fixatives for retrospective immunohistochemical and histopathological studies of viral infection in autopsy and biopsy materials. PMID- 3027433 TI - Anti-fibrotic effect of malotilate on liver fibrosis induced by carbon tetrachloride in rats. AB - The effect of malotilate on liver fibrosis of rat induced by carbon tetrachloride (CCl4) was studied. CCl4 was given subcutaneously (1.5 ml/kg) to male Wistar rats twice a week for 11 weeks. Administration of malotilate (100 mg/kg) 5 times a week was started in the 5th experimental week and continued thereafter. The biochemical parameters in serum such as concentration of total protein and transaminase activities were improved by malotilate administration. The livers treated with CCl4 only were atrophic and markedly nodular, and histologically, severe collagen deposition and pseudolobular formation were observed. However, the livers treated with CCl4 and malotilate showed only slight accumulation of collagen fibers although hypertrophic and fatty metamorphosis was observed. Immunocytochemically, type I and type III collagens were stained in livers from rats treated with CCl4 only and rats treated with CCl4 and malotilate, but stainability was rather weak in the latter. The increased amount of hydroxyproline in the liver and that excreted into urine were markedly suppressed by malotilate administration. The levels of protocollagen prolyl hydroxylase activity in the liver were almost the same in CCl4-treated and CCl4 and malotilate-treated rats. However, the levels of collagenolytic enzyme activity were slightly higher in the latter. From these results, anti-fibrotic action of malotilate was clear and involvement of enhanced collagenolytic enzyme activity was suggested. PMID- 3027434 TI - [Immunoglobulin synthesis of lymphocytes in patients with idiopathic nephrotic syndrome]. PMID- 3027435 TI - [Studies of human renal adrenergic receptors]. PMID- 3027436 TI - [A case of chronic renal failure associated with severe peripheral neuropathy]. PMID- 3027437 TI - Studies on mechanisms of antinephritic action of SA-446 an angiotensin I converting enzyme inhibitor (1). A comparison with actions of spironolactone, kallidinogenase and saralasin. AB - The present study was made to clarify the mechanisms of the antinephritic action of SA-446, an angiotensin I converting enzyme inhibitor, on crescentic-type anti GBM nephritis in rats as compared to the actions of spironolactone (an antialdosterone agent), kallidinogenase (a kallikrein agent) and saralasin (an angiotensin II antagonist). SA-446 (25 mg/kg/day, p.o.) had a tendency to reduce the urinary protein excretion and plasma urea nitrogen content. In addition, this drug remarkably inhibited not only glomerular histopathological changes (i.e., crescent formation, the adhesion of capillary walls to Bowman's capsule and fibrinoid necrosis) but also the elevation of blood pressure. Spironolactone (25 mg/kg/day, p.o.) and kallidinogenase (25 KU/day, i.m.) also showed beneficial effects on glomerular histopathological changes and hypertension, although both drugs were not as effective as SA-446. However, saralasin (72 micrograms/day, s.c.) caused a marked aggravating action on this nephritis. This nephritic model showed a marked low activity of plasma renin all through the 40 day experimental period. In this model, the urinary aldosterone excretion was increased, in spite of the decrease in plasma renin activity. SA-446 and kallidinogenase significantly inhibited the decrease in plasma renin activity and the increase in urinary aldosterone excretion. Spironolactone inhibited only the increase in the aldosterone excretion. However, saralasin decreased the plasma renin activity under the control level and strongly increased the urinary aldosterone excretion (about 1.8 times the control level on the 20th day). These results suggest that the antinephritic effect of SA-446 may be related to the antihypertensive action and the increase in renal blood flow through activation of the kallikrein-kinin and prostaglandins systems. PMID- 3027438 TI - Pharmacological characterization of presynaptic alpha-adrenoceptors in the modulation of the 5-hydroxytryptamine release from vascular adrenergic nerves in the rat. AB - The role of presynaptic alpha-adrenoceptors in modulation of the 5 hydroxytryptamine (5-HT) release from vascular adrenergic nerves was investigated in the perfused mesenteric vascular bed of the rat. After treatment with 5-HT (10 microM) for 15 min, the vasoconstrictor response to periarterial nerve stimulation (PNS, 4 to 16 Hz, 2 msec in duration for 30 sec) was greatly potentiated without significantly affecting the pressor response to exogenously administered noradrenaline (0.5 nmol). The potentiating effect was more pronounced at low frequencies of PNS (4 and 8 Hz). The potentiation of the pressor response to PNS after 5-HT treatment did not occur in the presence of LY53857 (0.01 microM), a selective 5-HT2 receptor antagonist. The enhanced pressor response to PNS seen after 5-HT treatment was further exaggerated in the presence of clonidine (0.1 and 1 microM), a preferential alpha 2-adrenoceptor agonist, while methoxamine (1 and 10 microM), a selective alpha 1-adrenoceptor agonist, did not affect the enhanced PNS response. This effect of clonidine was more pronounced in low frequencies of PNS (4 and 8 Hz) and was abolished by LY53857 (0.01 microM). In the perfused mesenteric vascular bed labelled with [3H] 5-HT, PNS (8 Hz) evoked an increase of tritium efflux in the perfusate. The PNS evoked tritium efflux was facilitated by yohimbine (0.1 to 1 microM), an alpha 2 adrenoceptor antagonist, and prazosin, a selective alpha 1-adrenoceptor antagonist, at a high concentration (1 microM), while LY53857 (0.01 to 0.1 microM) and a low concentration of prazosin (0.1 microM) had no effect on the tritium efflux. Clonidine (0.01 to 1 microM) produced a dose-dependent increase of PNS-evoked tritium efflux, while methoxamine (0.1 to 10 microM) was without effect. The monoamine uptake inhibitor, cocaine (10 microM) produced a significant inhibition of the PNS-evoked tritium efflux. The effects of clonidine and cocaine on the PNS-evoked tritium efflux were antagonized by yohimbine (1 microM). These results suggest that the release of 5-HT from adrenergic nerve endings by PNS is modulated by presynaptic alpha 2-adrenoceptors. PMID- 3027439 TI - [Superoxide production by neutrophils in patients with interstitial and other lung diseases]. PMID- 3027440 TI - Low-sodium resistant non-adrenergic inhibitory neurotransmission in the guinea pig duodenum. AB - The evoked inhibitory potentials (i.p.s) in the longitudinal smooth muscle cells of the guinea-pig duodenum were recorded intracellularly. The i.p.s were not blocked by adrenergic blocking agents, guanethidine (10(-6) g/ml), propranolol (10(-6) g/ml) and phentolamine (10(-6) g/ml), and atropine (10(-6) g/ml). Tetrodotoxin (10(-7)-10(-6) g/ml) abolished the non-adrenergic non-cholinergic i.p.s evoked by single or repeated stimulation without changes in the resting membrane potential of the longitudinal smooth muscle. In the presence of atropine (10(-6) g/ml), acetylcholine (5 X 10(-9)-7 X 10(-7) g/ml) had no effect on the amplitude of the i.p.s d-tubocurarine (2 X 10(-5) g/ml) reduced the amplitude of the i.p.s and produced a small depolarization in the muscle membrane. The longitudinal muscle membrane was slightly depolarized by Li-solution and the i.p.s could be evoked in the Li-solution for a long period, accompanying with the slight decrease in the amplitude of the i.p.s. In the choline-solution, the depolarization of the muscle membrane and the slight increase in the amplitude of the i.p.s were obtained. The i.p.s were abolished in the Ca2+-free choline solution. In the sucrose-solution, the decrease of the resting membrane potential of the longitudinal smooth muscle was observed and the amplitude of the i.p.s was gradually reduced during the perfusion of the sucrose-solution. Chloride deficiency had no considerable effect on the amplitude of i.p.s. The results obtained suggest that the non-adrenergic non-cholinergic inhibitory neurotransmission is low-sodium resistant and the excitation-secretion coupling in the non-adrenergic non-cholinergic inhibitory nerves is mainly dependent on the external calcium ions but not sodium ions. PMID- 3027441 TI - B-cell maturation stages of Burkitt's lymphoma cell lines according to Epstein Barr virus status and type of chromosome translocation. AB - This study addressed the possible relationship between B-cell maturation stage of Burkitt's lymphoma (BL) cell lines and Epstein-Barr virus (EBV) status, ethnic group, or type of chromosome translocation. Fifty-seven cell lines obtained at the International Agency for Research on Cancer from 51 patients were studied. Cytogenetic analyses of 54 cells lines were available. Cell size, surface immunoglobulins (sIgs), cytoplasmic immunoglobulins (cIgs), mouse red blood cell receptors, and reactivity with various monoclonal antibodies were assessed. Immunoglobulin (Ig) class secretions were measured in the supernatant of 2- and 5 day cultures from 33 cell lines, with the use of a sensitive enzyme-linked immunosorbent assay technique. From this study, BL appears to cover a broad range of the B-cell differentiation sequence, since the following Ig phenotypes were observed: null cells (sIg-, cIg-), large pre-B-cells (intracytoplasmic mu chains), small B-cells (sIg+, cIg-), and various types of secreting B-cells (sIg+, cIg+). Among the latter, various patterns of cIg could be defined (perinuclear, paranuclear, and vesicular). B-cell maturation stages were correlated with the amount of secreted Ig. In sIg+ cell lines, different classes of Ig were found: 35 IgM, 10 IgM plus IgD, 4 IgG, and 1 IgA. None of the different monoclonal antibodies used was specific to a precise stage of maturation. The stages of maturation were correlated with neither the type of chromosome translocations of BL nor the presence of EBV genome, but the most immature cell lines were all EBV positive and most of them originated from African patients. In contrast with acute lymphoblastic leukemia, the common acute lymphocytic leukemia antigen (CD10) was expressed on nearly all BL cell lines of intermediate maturation stages but only on half of the pre-B ones. In addition, none of the cell lines tested was found to react with CD5 antibodies, which recognize most of the chronic lymphocytic leukemia of the same stage of maturation as that in the B-lymphocyte lineage. PMID- 3027442 TI - Retinoic acid-induced changes in 1 alpha,25-dihydroxyvitamin D3 receptor levels in tumor and nontumor cells derived from rat bone. AB - The inducibility of 1 alpha,25-dihydroxyvitamin D3 [1,25-(OH)2D3] binding by all trans-retinoic acid (RA) was examined in tumor-derived clonal bone cell lines, in established clonal cell lines derived from normal embryonic bone, and in cultured bone cell populations freshly isolated from 18- and 21-day fetal and 5-day-old neonatal rat calvaria. Levels of 1,25-(OH)2D3 binding were determined using a single saturating dose (84 pM) of 3H-labeled 1,25-(OH)2D3. Bone-derived tumor cell lines (ROS 17/2.8, ROS 17/2, RCJ 3.2T.1, RCJ 3.2.4.1CAM, RCJ 3.2CE2.1) possessed high basal levels of binding and showed increases in 1,25-(OH)2D3 binding after culture for 24 hours in the presence of 10(-5) M RA. The non-tumor derived established bone cell lines (RCB 2.2A, RCB 2.2B, RCB 2.2C, RCB 2.2D) showed low basal 1,25-(OH)2D3 binding levels and no change in response to RA, while first subcultures of bone cell populations derived from fetal and neonatal rat calvaria showed decreased 1,25-(OH)2D3 binding, following similar treatment with RA. In representative cell populations, the dose dependency of the RA effect was established. The observed differences in response to RA in the cell lines tested seem to be dependent on whether the cells originated from normal or tumor tissue. PMID- 3027444 TI - [Anti-arrhythmic and anti-ischemic action of guanosine-5'-monophosphate]. PMID- 3027443 TI - [Age dependent quantitative evaluations of sensitivity changes in beta adrenergic receptors to isoprenaline and propranolol]. PMID- 3027445 TI - [Herpesvirus infections in children with acute lymphatic leukemia]. AB - 33 children with acute lymphatic leukemia and 33 healthy controls were longitudinally studied for herpesvirus infections. Active herpes simplex-virus, varicella-zoster virus and cytomegalovirus (CMV) infections were more frequent in patients than in controls. CMV and Epstein-Barr virus infections were often inapparent or associated with infections of the upper respiratory tract. Analysis of serological datas revealed a coincidence of active CVM infection and lethal course of the leukemia. This may be a result of the immunodeficiency in leukemia patients caused by disease and therapy. An additional influence of the immunosuppressive effect of active CMV infections on the course of the disease is discussed. PMID- 3027446 TI - [Catecholamine behavior, adrenoreceptor density of intact cells and sensitivity to catecholamines in a patient with orthostatic hypotension]. AB - We evaluated sympathetic nervous system function in a patient with primary orthostatic hypotension. Plasma catecholamine levels--except for dopamine levels- and urinary catecholamine excretion were decreased, alpha-adrenoreceptor responsiveness to noradrenaline and beta-adrenoreceptor responsiveness to isoproterenol were increased according to increased beta-2-adrenoreceptor density on intact polymorphonuclear leukocytes. Alpha-2-adrenoreceptor density on intact platelets and adrenaline-induced platelet aggregation in vitro, however, were unchanged. We evolved a therapeutic regimen with fludrocortisone, propranolol, and dihydroergotamine that allowed the patient to resume nearly a regular degree of mobility. PMID- 3027447 TI - [Morphological and biochemical analysis of the organs and tissues of rats following a 30-day exposure to increased gravitational forces of 1.1 and 2.0 G]. AB - During 30 days rats were centrifuged at 1.1 and 2.0 G. On centrifugation day 30 the rats showed body mass losses, decrease of plasma ACTH, activation of the renin-angiotensin-aldosterone system (RAAS) and ultrastructural changes in the mossy fiber terminals in the nodulus cortex which were indicative of the state of excitation (at 1.1 G) or excess excitation (at 2.0 G) in the system of the utriculus receptor cell and vestibular ganglion neuron (RCN). On the 2nd day after centrifugation the ultrastructural changes in the terminals pointed to a lower activity of the RCN system which was below the physiological norm. As compared to centrifugation day 30, the RAAS became more active on the 2nd day of recovery. On the 7th day of recovery (after centrifugation at 1.1 G) the RCN ultrastructure, RAAS and ACTH concentrations returned to the normal. The general trends of the RAAS and RCN changes seen on the 2nd day of recovery and identified by other authors at an acute stage of adaptation to microgravity suggest that the data obtained on the 2nd day of recovery may be used to analyze certain effects which develop during an acute stage of adaptation to microgravity in mammalian organs and systems responsible for the perception of modified gravity and their adaptation to a new level of gravity. PMID- 3027449 TI - Establishment of human fibroblast cell lines with lysosomal enzyme deficiency by transformation with origin-minus SV40 DNA. PMID- 3027448 TI - Biochemical phenotyping of a single sibship with both cystinosis and Fabry disease. AB - Two lysosomal storage diseases, cystinosis and Fabry disease, were diagnosed clinically in different members of a single sibship. The possibility that the affected sister and brother might have related disorders with disparate manifestations was pursued. The four principal family members were tested for heterozygote status with respect to serum and leukocyte alpha-galactosidase A activity, urinary trihexosylceramide excretion, and the capacity to engage in cystine counter-transport across leukocyte lysosome membranes. Results were consistent with classical autosomal recessive inheritance in the case of cystinosis and X-linked inheritance for Fabry disease, confirming that this family represents an example of two rare disorders occurring in the same sibship. PMID- 3027450 TI - Lack of acid sphingomyelinase in the mitochondria-lysosome fraction of brain from Niemann-Pick mice. PMID- 3027451 TI - Structure of the 5' end region of the human argininosuccinate synthetase gene. PMID- 3027452 TI - Mortality of asbestos cement workers using almost exclusively chrysotile fibre. PMID- 3027453 TI - Superoxide radicals and hemorrhagic shock: are intravascular radicals associated with mortality? AB - Superoxide radicals (SORs) are formed during hemorrhagic shock. However, the association between SOR formation and mortality remains undefined. Would neutralization of SORs during shock improve survival? Superoxide dismutase (SOD) was covalently linked to Ficoll to prolong vascular persistence and ensure ongoing neutralization of intravascular SORs. Six groups of Sprague-Dawley rats were studied. Rats were exsanguinated to a mean arterial pressure of 30 Torr, sustained for 60 min. Ficoll linked superoxide dismutase (SOD-F), 5000 Units + catalase, 150 Units, was suspended in 1 ml of saline and injected following the 60-min hypotensive period. Resuscitation followed for appropriate groups. Pretreated SOD-F rats received a 1-ml bolus prior to exsangination. Pretreatment with SOD-F did not improve survival in non-resuscitated groups despite improvement in vascular circulation (P greater than 0.10, NS). In the standard non-resuscitation group, there is also no improvement (P greater than 0.10, NS). Resuscitation with or without SOD-F significantly alters survival rates (P less than 0.025) with respect to non-resuscitated controls. However, application of SOD-F in conjunction with resuscitation does not improve survival with respect to resuscitation controls (P greater than 0.10, NS). Although SOD-F has a vascular half-life advantage over SOD, the improvement in survival is statistically non significant. Intravascular SORs do not appear to be associated with mortality. SOR release and consequent damage to the vasculature may be preceded by a more significant intracellular damage. Intracellular SOR damage and its association with mortality in hemorrhagic shock remains an open issue. PMID- 3027454 TI - Effect of dietary supplementation with vitamin A on arterial healing in rats. AB - The effect of dietary supplementation with vitamin A on the healing of rat aortas was studied. Rats were divided into two groups: rats fed a standard chow and rats fed the chow supplemented with 150 IU vitamin A palmitate/g diet. Seven rats from each group were fed with the above diets for 7 days, killed with ether, and the abdominal aorta excised and assayed for hydroxyproline content and collagenase activity. Ten rats from each group were fed for 7 days on the above diets and then underwent a longitudinal aortotomy which was sutured with prolene sutures under pentobarbital anesthesia. The rats were maintained on their respective diets for 7 days and then killed with ether, their abdominal aorta was excised and both the segment with the arteriotomy and the adjacent distal normal segment were analyzed for hydroxyproline content and collagenase activity. Seven rats from each group were fed with the above diets and then underwent transverse division of the abdominal aorta and reanastomosis using nylon sutures under pentobarbital anesthesia. Rats were maintained on their respective diets throughout the postoperative period. Seven days later, all rats were killed with ether and the bursting strength of the aortic anastomoses was measured. The results showed that vitamin A supplementation in non-operated animals had no significant effect on aortic hydroxyproline content or collagenase activity. In rats undergoing longitudinal aortotomy and suture, there was a significant increase in hydroxyproline content both at the healing arteriotomy and at the adjacent non-wounded aorta in the vitamin A-supplemented group. There was also a significant increase in bursting strength of the healing aortic anastomosis in the vitamin A-supplemented rats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3027455 TI - Transduction of ACTH signal from plasma membrane to mitochondria in adrenocortical steroidogenesis. Effects of peptide, phospholipid, and calcium. AB - The conversion of cholesterol to pregnenolone by adrenocortical mitochondria is the rate-limiting step in steroidogenesis. This process is stimulated dramatically by the action of ACTH through the sequential reactions, in which adenyl cyclase, cAMP-dependent protein kinase, cholesterol esterase and ribosomal protein synthesis are all involved. The de novo synthesized protein, the so called labile protein with a half-life of approx 10 min, is believed to stimulate the cholesterol side chain cleavage reaction by an unknown mechanism. Available evidence indicates that the electron on transfer reaction from NADPH to P-450scc is mediated rapidly by adrenodoxin reductase and p-450 scc. In addition, these redox components are inactivated slowly with a half-life of 3.5 days after hypophysectomy. It is known that the corticoid output from adrenocortical cells starts within 5 min and reaches the maximum after 10-15 min of ACTH administration to animals. One can assume that under normal physiological conditions, both O2 and NADPH are not limiting. Additionally, mitochondrial inner membranes are poor in cholesterol. In this context, the availability of substrate cholesterol to P450scc is the most likely candidate for the regulatory mechanism. PMID- 3027457 TI - Steroid dynamics in the normal and carcinomatous mammary gland. AB - As an approach to the study of the origin of estrogens in breast cancer tissue, the concentration of estrogens and their androgen precursors, as well as aromatase and 17 beta-dehydrogenase activities in normal glandular (GL) and cancerous (CA) breast tissue were determined and correlations with plasma levels and/or receptor status were studied. In both normal GL and CA breast tissue, steroid concentrations were significantly higher than plasma conc., except for dehydroepiandrosterone sulphate (DHEAS), estrone sulphate (E1S) and testosterone (T). Androgen conc. were lower, but estrogen conc. were higher in CA than in GL breast tissue. Estradiol (E2) conc. was positively correlated with the E2R conc., mean E2 conc. corresponding to an estimated E2R occupancy of about 25%. Aromatase and 17 beta-hydroxysteroid dehydrogenase (E2DH) (E2----E1) activities were observed in all breast CA and GL tissues, aromatase accounting probably only for a small fraction of tissue estrogens. E2DH, but not aromatase activity, was significantly higher in E2R+ than in E2R- tissues and was negatively correlated with tissue dehydroepiandrosterone (DHEA) and DHEAS conc.; the latter two steroids are non competitive inhibitors of E2DH which inactivates E2 to E1. CONCLUSION: in both normal and carcinomatous breast tissue, conc. of E1 and E2 are significantly higher than in plasma, suggesting either uptake or local synthesis. As to the latter, aromatase activity accounts probably only for a minor fraction of the tissue estrogens. Breast CA tissue has higher aromatase and E2DH activity than normal glandular tissue, E2 conc. and E2DH activity being higher in E2R+ hormone sensitive, tumors than in E2R-tumors. Tissue conc. of DHEA(S) which inhibits oxidative inactivation of E2, is negatively correlated with E2DH activity and may have an important modulating role in intratissular estrogen metabolism. PMID- 3027456 TI - Determination of urinary lignans and phytoestrogen metabolites, potential antiestrogens and anticarcinogens, in urine of women on various habitual diets. AB - Recently two groups of compounds with diphenolic structure, the lignans and the isoflavonic phytoestrogens, were detected and identified in human urine and other biological fluids. These compounds are of great biological interest because they exhibit both in vitro and in vivo weak estrogenic and sometimes also antiestrogenic activities and many plant lignans have been shown to have anticarcinogenic, antiviral, antifungal and other interesting biological effects. The compounds found in relatively large amounts (10-1000 times more than estrogens) in urine are modified by intestinal bacteria from plant lignans and phytoestrogens, which are present in fiber-rich food such as grain and beans. They bind with low affinity to estrogen receptors and preliminary results suggest that they may induce production of sex hormone binding globulin (SHBG) in the liver and in this way may influence sex hormone metabolism and biological effects. Five compounds, the lignans enterolactone (Enl), enterodiol (End) and the isoflavonic phytoestrogen metabolites daidzein (Da), equol (Eq) and O desmethylangolensin (O-Dma) were measured in urine by gas chromatography-mass spectrometry (selected ion monitoring) using deuterated internal standards in 5 groups of women (total number 53). The members of three dietary groups (omnivores, lactovegetarians and macrobiotics) were living in Boston and of two groups in Helsinki (omnivores and lactovegetarians). Until now measurements have been carried out in 94 72-h samples. The highest mean excretion of the most abundant compound, enterolactone, was found in the macrobiotic group and the lowest in the omnivoric groups. Total mean 24-h excretion of enterolactone was 17,680 nmol in the macrobiotics, 4,170 nmol in the Boston lactovegetarians, 3,650 nmol in the Helsinki lactovegetarians, 2,460 nmol in the Helsinki omnivores and 2,050 nmol in the Boston omnivores. The other diphenols followed approximately the same pattern. In an earlier study the lowest excretion of enterolactone (1,040 nmol/24 h) was found in a group of postmenopausal apparently healthy breast cancer patients living in Boston. It is concluded that further studies are necessary to elucidate the possible role of these compounds in cancer and other diseases. However, the evidence obtained until now seems to justify the conclusion that these compounds may be among the dietary factors affording protection against hormone-dependent cancers in vegetarians and semivegetarians. PMID- 3027459 TI - Primary malignant melanoma of the lung. AB - A case of primary malignant melanoma of the lung in a 42-year-old white female is presented. A 6 cm localized mass in the left lower pulmonary lobe was discovered on a thoracic spine X-ray done for unrelated chronic back pain. The patient underwent left lower lobectomy for cure. Multiple detailed physical examinations did not disclose any primary lesions. Other cases from the English literature are reviewed briefly. PMID- 3027458 TI - Malignant fibrous histiocytoma of the breast with axillary lymph node involvement. AB - This case history describes a 70-year-old female patient with a malignant fibrous histiocytoma (MFH) of the breast with secondary involvement of the skin and axillary lymph node metastases. We think that in the case of a MFH of the breast a radical or modified radical mastectomy should be pursued instead of a simple mastectomy. PMID- 3027461 TI - The immunoregulatory effects of ginsenoside in vivo and in vitro. PMID- 3027460 TI - Effects of acupuncture on left ventricular function, microcirculation, cAMP and cGMP of acute myocardial infarction patients. PMID- 3027462 TI - 13-cis retinoic acid treatment of myelodysplastic syndromes. AB - Of eight patients with primary myelodysplastic syndrome (MDS) treated with Roacutane (13-cis retinoic acid, Roche, Basel, (13-RA)) 20 mg/m2 for 6 weeks and an additional 100 mg/m2 for 4 weeks (3 patients), 4 responded either with a slight increase in peripheral blood neutrophil count or a decrease in myeloperoxidase deficient neutrophils. In agar cultures 2 patients showed a concurrent increase in growth of day 11 colonies and clusters. In 2 of the patients a decrease in the number of immature bone marrow cells positive for the myeloid antibody anti-My7 was observed. Only minor alterations were seen in natural killer cell activity. In 4 patients showing clonal chromosomal abnormalities before treatment a disappearance of minor clonal abnormalities during treatment was observed, and in 3 chromosomal abnormalities reappeared after cessation of therapy. Even though the overall impact of 13-RA on the hematopoietic system was minor, the increase in myeloperoxidase normal granulocytes in the blood and the decrease in My7 positive cells and in clonal chromosomal abnormalities warrants further interest in this agent as a treatment modality in MDS. The side effects, especially experienced by the patients receiving 100 mg/m2, were, in spite of symptomatic treatment, of such a degree that only low dose treatment (10-20 mg/m2) administered for prolonged periods of time (3-6 months) would seem recommendable. PMID- 3027464 TI - Magnesium-nucleic acid conformational changes and cancer. AB - The magnesium interaction with the mononucleotide guanosine-5'-monophosphate disodium salt (5'-GMP Na2 or G5'p) has been studied by Fourier transform infrared spectroscopy. The results have been compared with previously obtained proton nuclear magnetic data on the same system as well as other similar systems. The spectra here show that the natural conformation of the mononucleotide changes significantly in the presence of normal concentrations of magnesium chloride (approximately 10(-3) M) from the predominantly C2'-endo,anti into C3'-endo,anti conformation of the sugar ring, and the phosphate group becomes virtually gauche gauche. The marker bands for C2'-endo,anti and C3'-endo,anti are shown near 820 cm-1 and 803 cm-1, respectively. The infrared spectroscopic results found here indicate that the unit Mg(H2O)2+5 is coordinated at the N7 site of the base guanine and hydrogen bonded via the water molecules to the carbonyl C6 = O and to the phosphate negative oxygens. This type of bonding is very similar to the binding of the antitumor drug cis-platinum, which is chemically (covalently) bound to the N7 site of guanine in the same molecule or in nucleic acids. PMID- 3027463 TI - Elevated serum neopterin levels in sarcoidosis. PMID- 3027465 TI - Maintenance of fetal human pancreatic beta cells in tissue culture. AB - Large quantities of viable human islet tissue (beta cells) are required for transplant and for investigations of the autoimmune basis of Type I diabetes. Fetal pancreas offers a potential advantage over other possible sources of beta cells in that it retains some capacity for growth in vitro. We have cultured a total of 45 human pancreata from fetuses of gestational ages from 18 to 23 weeks. Each pancreas was obtained within minutes after delivery and usually cultured within 30 minutes. Pancreata were dispersed and cultured for up to 32 days. Maintenance and growth of the beta cells was assessed by the content of insulin in extracts of cultured tissue. As has been reported by others, fetal human beta cells survived in vitro for over 4 weeks. In three experiments in which a direct comparison was made, collagenase digestion of the fetal pancreas resulted in a significantly greater loss of insulin content compared to minced tissue cultured without digestion. Storage of three pancreata in medium overnight at 4 degrees C significantly reduced the insulin content of the pancreas compared to pancreata cultured immediately. During culture, the majority of the beta cells (based on insulin content) were found in small, macroscopic clumps attached to the surface of the culture dish, and surrounded by a nearly confluent monolayer of fibroblastoid cells. There was a marked decrease in the insulin content of the tissue during culture, most of it (to less than 25% of the original) occurring over the first 4-6 days of culture.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3027466 TI - Autocrine growth of human B lymphocytes: maintained response to autostimulatory factors is the special feature of immortalization by Epstein-Barr virus--a hypothesis. AB - The autocrine growth profile of human B lymphocytes transformed with Epstein-Barr virus (EBV) was found to comprise three distinct components: a B-cell growth factor (BCGF); an interleukin-1 (IL-1)-like activity; an activity requiring cell to-cell contact for its action. Observations on the inhibition of the EBV carrying Daudi lymphoma line by alpha-interferon indicated that loss of response to these autostimulatory factors was underlying growth cessation. Furthermore, a putative receptor for BCGF was found to be down-regulated on B cells stimulated with non-transforming mitogens but constitutively expressed following EBV transformation. Taken together with recent evidence that normal B cells produce autostimulatory factors, these findings suggest that the special feature of autocrine growth by EBV-immortalized cells is a maintenance of what should normally be a transient phenotype, possibly through deregulation of receptor expression. This hypothesis is discussed. PMID- 3027467 TI - Role of alpha-adrenergic receptors in cartilaginous and non-cartilaginous human airways. AB - An imbalance between the density of alpha- and beta-adrenergic receptors in airway smooth muscle has been suggested as one of the potential underlying mechanisms of bronchial asthma. Therefore, the present study was conducted to examine the presence and functional behavior of alpha-adrenoceptors in isolated human bronchi and bronchioles. Human airway tissues did not show any spontaneous tone during equilibration in physiological solution. Sensitivity of non cartilaginous airways (8th generation and smaller) to histamine was approximately 3 times more than cartilaginous (2nd-7th generation) airways. In the presence of beta-adrenoceptor blockade, norepinephrine (3 X 10(-4) M) evoked approximately 14 40% of maximum histamine contraction in cartilaginous and non-cartilaginous airways from three donors. When tone in these tissues was elevated by pretreatment with histamine (10(-5) M), leukotriene D4 (10(-10) M), carbachol (1 X 10(-6) M-3 X 10(-6) M) or KCl (10(-2) M), the sensitivity to norepinephrine was potentiated. Airway tissue from one donor was found to be highly reactive to norepinephrine and the maximum contractions were 118 and 145% of maximum histamine contraction for cartilaginous and non-cartilaginous airways, respectively. Time to reach maximum contraction in cartilaginous airways was 80 min compared to 21 min for non-cartilaginous airways from the same donor. These data suggest that human airways contain contractile alpha-adrenoceptors in addition to relaxant beta-adrenoceptors, which haven been reported previously in the literature. Though activation of alpha-adrenoceptors induces contractions in human airway smooth muscle, the potency of norepinephrine for alpha-adrenoceptors in these airways is low compared to that reported for vascular smooth muscle. PMID- 3027468 TI - The role of H1- and H2-receptors in the modulatory effects of histaminergic agents on adrenergic neurotransmission in rat vas deferens. AB - The role of the H1- and H2-receptors in the modulatory effects of histamine and other histaminergic drugs on smooth muscle adrenergic neurotransmission was studied on isolated preparations from rat vas deferens. The typical biphasic action (potentiation by low concentrations and inhibition by high concentrations) of histamine and some H1- or H2-agonists on vas deferens contractions induced by low frequency electrical stimulation (ES) was markedly changed by H1- or H2 antagonists. After blockade of H1-receptors, the potentiating effect of histamine and the histaminergic agonists was diminished or even reversed. The inhibition of the smooth muscle contractions by the drugs tested was stronger after H1 antagonists. Blockade of H2-receptors usually enhanced the potentiating effect of the histaminergic agonists on ES-evoked vas deferens contractions. The inhibitory action of histamine and the histaminergic agents tested was decreased or even reversed after H2-receptor blockade. These results confirm the presence of H1 excitatory and H2-inhibitory receptors whose activation or inhibition can modulate adrenergic neurotransmission in vas deferens. PMID- 3027469 TI - A study on the antagonism of the intestinal inhibitory effects of morphine and clonidine by yohimbine in mice. AB - In mice, clonidine administered subcutaneously caused a dose-dependent inhibition of the intestinal motility as assayed by the movement of a charcoal meal. This inhibitory effect of clonidine was antagonized dose-dependently by prior subcutaneous or intracisternal administration of yohimbine. However, yohimbine given intracerebroventricularly was ineffective in antagonising the intestinal inhibitory action of clonidine. Clonidine administered centrally, either intracisternally or intracerebroventricularly, caused a dose-dependent inhibition of intestinal motility. Clonidine given by the intracisternal route appeared to be more effective than by the intracerebroventricular route. Centrally administered clonidine was antagonized by prior subcutaneous administration of yohimbine. The antagonism was related to the doses of yohimbine given. Subcutaneously administered morphine caused a dose-dependent inhibition of intestinal motility and this effect was antagonized by prior subcutaneous administration of yohimbine. However, administration of yohimbine centrally, either intracisternally or intracerebroventricularly, did not affect the intestinal inhibitory action of morphine. On the other hand, morphine injected centrally, either intracisternally or intracerebroventricularly, dose-dependently inhibited the motility of the intestine; such inhibitory action was antagonized by prior subcutaneous administration of yohimbine. The present study suggests that clonidine inhibits intestinal motility at both central and peripheral sites through alpha 2-adrenoceptors. Morphine also inhibits intestinal motility by both central and peripheral mechanisms but it appears that yohimbine only modifies the peripheral aspect of morphine's action. PMID- 3027470 TI - [Preventive and simultaneous operations of the abdomen]. AB - Prophylactic and elective operations have the same risk. They require detailed and extensive information of the patient and documentation thorough medical reasoning. Without preoperative planning prophylactic operations are justified in severe conditions. Simultaneous operations increase the risk in case they represent major procedures or in septic interventions. Simultaneous surgical interventions of the stomach and the biliary tract imply a high morbidity (13% pulmonary embolism and pneumonia) and a high lethality (15%). In simultaneous operations the fastest of the skilled surgeons is the one who is most successful. PMID- 3027471 TI - [Surgical indications in small-cell bronchial cancer]. AB - From a total of 1,366 patients with bronchial carcinoma, all selected for an operative procedure, we found 101 patients with small cell carcinoma. For out of 7 of those with small cell carcinoma only mediastinoscopy revealed histological proof of the carcinoma. Thus mediastinoscopy proves to be a valid means for preoperative evaluation of intrathoracic growth of a tumor. PMID- 3027472 TI - [Oncologic viewpoints of surgical and internal medicine cooperation]. AB - Chemotherapy is the treatment of first choice in small cell lung cancer. Retrospective analysis and preliminary results of prospective studies demonstrate that an additional local treatment will increase the incidence of two years recurrence free survival. This role would be taken over by surgical resection for patients with the very limited stages I and II. Surgery has the advantage as compared to radiotherapy of exact exploration of the thorax and analysis of lung tissue after chemotherapy. Furthermore side effects of radiotherapy to these organs would be avoided. Because of the higher possibility of distant metastases local surgery should be performed as an adjuvant procedure to chemotherapy. PMID- 3027473 TI - [Value of surgery in the treatment of small-cell bronchial cancer]. AB - As a result of strict cooperation between oncologists, surgeons and radiotherapists we developed a comprehensive combination therapy for small cell lung cancer (limited disease): Stage I, II: radical tumour resection; chemotherapy (Cohen); cerebral radiation prophylaxis; locoregional radiotherapy; Stage III: Chemotherapy; cerebral radiation prophylaxis; lung resection in an extension required by the initial tumour; radiotherapy (local). The preliminary results in 15 patients are encouraging. PMID- 3027474 TI - [Indications and results of the surgical treatment of small-cell bronchial cancer]. AB - From 1973 to 1983 170 patients underwent surgical treatment of small cell lung cancer. In 2/3 of the cases we performed lobectomy resp. bilobectomy including bronchoplastic and arterioplastic resections, the other third of patients underwent pneumonectomy. 26 patients (15%) died within 30 days. Postoperative mortality in patients with pneumonectomy was 17% and in patients undergoing lobectomy 7%. Since 1978 lethality rate dropped from 20% to 7%. Due to polychemotherapy, estimated time of survival (according to Kaplan and Meier) has increased significantly, adding up to 3-years- and 5-years-survival rates of 31% and 20% respectively. PMID- 3027475 TI - [Improved regional myocardial function after aortocoronary bypass surgery noninvasive studies using the first-pass technic]. AB - 50 patients after coronary artery bypass grafting were matched with 50 not operated patients. After a follow-up period of four years they were investigated by first pass radionuclide angiography. Revascularized myocardial areas showed significantly higher regional ejection fraction during exercise and in territories with prior myocardial infarction. The results proved the hemodynamic benefits of coronary artery bypass grafting even in territories with poor myocardial function. PMID- 3027476 TI - [Granular cell tumor of the orbits. Diagnosis and therapy]. AB - Report on two cases of tumours in the retroorbital space. The histological examination showed them to be granular cell myoblastomas (Abrikossoff), which in one case was bilateral. Tumours within the orbital space are rare. It is necessary in every case to make a biopsy to clarify the type of tumour because some of them are malignant. Computed tomography cannot differentiate between benign and malignant tumours in the orbita. PMID- 3027477 TI - [Unusual location of a granular cell myoblastoma (Abrikosov tumor)]. AB - A case of an unusual location of a granular cell myoblastoma is reported. This tumour is considered to be of neurogenic origin and has been found in different areas of the head and neck. A profound description of the histologic features is provided as well as a remark on the differential diagnosis and a review of the appropriate literature. PMID- 3027478 TI - Complications in the management of large glomus jugulare tumors. AB - This study analyzes the complications encountered in the surgical treatment of 17 patients with large glomus jugulare tumors. All 17 patients sustained either new cranial nerve palsies or exacerbation of preexisting palsies. These involved, in descending order of frequency, the facial nerve, the vagus and glossopharyngeal nerves, and the hypoglossal nerve. Postoperative palsies of the facial nerve involved 17 patients, as compared to 5 preoperatively. Fifteen patients had postoperative partial or complete paralysis of the vagus nerve as compared to eight preoperatively. Ten patients had postoperative palsies of the hypoglossal nerve as compared to six preoperatively. Other complications included CSF leak, meningitis, and wound infection. Aspiration and dysphagia were encountered postoperatively in 13 patients. Teflon injection of paretic vocal cords and cricopharyngeal myotomy effectively improved the ability to swallow and the quality of the voice. Prompt recognition and treatment of complications is essential for effective surgical management of large glomus jugulare tumors. PMID- 3027480 TI - Hormonal receptors in juvenile nasopharyngeal angiofibroma. AB - Specific thermostable androgen receptors were detected in the tissues of nasopharyngeal angiofibroma. The receptors seemed to be specific with high affinity toward DHT more than testosterone. No abnormalities in serum levels of DHT, testosterone, and estradiol-17 beta could be detected in the patients studied. A concept of pathogenesis of the tumor in relation to that reported in literature recently is interpreted in the text. PMID- 3027479 TI - Prostaglandins, leukotrienes, and other arachidonic acid metabolites in nasal polyps and nasal mucosa. AB - Prostaglandins (PGs) and leukotrienes (LTs) are known to play an important role in allergic inflammatory reactions. The triad of aspirin sensitivity, nasal polyposis, and asthma led us to suspect that PGs, LTs and other arachidonic acid metabolites may be involved in the pathogenesis of nasal polyps. The purpose of this study was to determine arachidonic acid metabolites and to measure concentrations of PGs and LTs in nasal polyps and nasal mucosa. Samples of nasal polyps and nasal mucosa were obtained at the time of polypectomies and nasal procedures. Metabolites of arachidonic acid in tissue were determined by incubation of tissue-homogenates with 14C-arachidonic acid and analyses with thin layer chromatography and high performance liquid chromatography (HPLC). Levels of PGE2, 6-keto-PGF1 alpha, thromboxane (Tx)B2, 15-hydroxyeicosatetraenoic acid (HETE), LTC4, LTB4 were measured by radioimmunoassay. The predominant arachidonic acid metabolite in both nasal polyps and mucosa with 15-HETE. The HPLC analysis showed that the predominant metabolite in nasal polyp was 15-HETE, especially in polyps from aspirin sensitive patients. Levels of 15-HETE and PGE2 were higher in polyps from patients with a history of allergy than from nonallergic patients. Levels of LTC4 and LTB4 in nasal polyps were determined. The findings of this study will help to explain biochemical basis of the pathogenesis of aspirin sensitive nasal polyps and to develop better medical treatment for them. PMID- 3027481 TI - Discriminative stimulus effects of N-allylnormetazocine in rats trained to discriminate a kappa from a sigma agonist. AB - Rats were trained to discriminate ethylketocyclazocine (EKC) from vehicle, phencyclidine (PCP) from vehicle, or ethylketocyclazocine from phencyclidine on a two-lever operant task with a fixed-ratio 30 schedule of food-reinforcement on the appropriate lever. The three groups were tested with the training drugs (i.e., EKC and PCP), N-allylnormetazocine (NANM), and the (+)- and (-)-isomers of N-allylnormetazocine. EKC produced EKC-appropriate responding in the EKC-vehicle group and in the EKC-PCP group; it produced vehicle-appropriate responding in the PCP-vehicle group. Similarly, PCP produced PCP-appropriate responding in the PCP vehicle group and in the EKC-PCP group but vehicle-appropriate responding in the EKC-vehicle group. The (+)-isomer of NANM produced PCP-appropriate responding in both the PCP-VEH and EKC-PCP groups, whereas the (-)-isomer produced EKC appropriate responding in the EKC-VEH and EKC-PCP groups. The results of this study demonstrate that rats can be trained to discriminate a kappa-agonist from a PCP/sigma-agonist and can differentiate these discriminative stimulus properties of other test compounds. These results also indicate that the (-)-isomer of NANM has kappa-agonist discriminative stimulus properties, whereas PCP/sigma-like effects are produced by the (+)-isomer. PMID- 3027482 TI - Effect of inhibition of Na+-K+ ATPase on the prostacyclin generation of cultured human vascular endothelial cells. AB - Prostacyclin (PGI2) generation of cultured human vascular endothelial cells (VEC) was observed coincidentally with the increase of 45Ca net influx. Ca ionophore A23187 enhanced not only PGI2 generation and 45Ca net influx but also 45Ca efflux. PGI2 generation was completely abolished by the pretreatment with Ca++ immobilizer, TMB-8. A Na+-K+ ATPase inhibitor, ouabain increased 45Ca net influx, but decreased 45Ca efflux, and enhanced PGI2 generation. These observation indicate that PGI2 generation of VEC may be regulated by not only Ca++ but also Na+, and it was suggested that enhanced PGI2 generation by ouabain might be derived from the increased cytosolic Ca++concentration by the decreased Ca++ efflux, and it was considered to be originated from the suppression of Na+-Ca++ exchange systems by the increased intracellular Na+ concentration via inhibition of Na+-K+ ATPase activity by ouabain. Enhancement of PGI2 generation of VEC by the increased ouabain like substances (OLS) in hypertension is suspected to be beneficial on the maintenance of vascular homeostasis. PMID- 3027483 TI - Decrease of mu opioid receptors in the brain and in the hypothalamus of the aged male rat. AB - Experiments have been designed in order to analyze whether the binding capability of mu opioid receptors in the brain of the male rat is modified by age. In a first experiment, the number of receptors (Bmax) and the constant of affinity (Ka) for the mu ligand 3H-dihydromorphine (3H-DHM) have been measured in the whole brain of male rats of 2, 15 and 22 months of age. In a second experiment the Bmax and the Ka for 3H-DHM have been evaluated in the hypothalamus of male rats of 2 and 22 months of age. In this experiment it was also investigated whether the administration of exogenous testosterone modifies the number and/or the affinity of the hypothalamic mu receptors. Serum levels of LH, FSH, prolactin and testosterone have been measured by specific RIAs. The results obtained show that: serum testosterone levels are significantly decreased in aged rats, while serum LH and FSH show only a small decline; serum prolactin is higher in old than in young animals; the number of mu receptors in the whole brain of 15 and 22 month old animals and in the hypothalamus of 22 month old rats is significantly lower than in the same tissues of young animals; the administration to old animals of testosterone, in doses able to bring back towards normal serum levels of testosterone, induces a decrease of LH and FSH, but has no effect on serum prolactin titers. Testosterone administration does not modify the number of hypothalamic mu opioid receptors, indicating that the decline of brain mu receptors in old animals is not the consequence of the physiological decline of testosterone secretion; in no instance the Ka for the mu ligand is significantly affected. PMID- 3027484 TI - High affinity binding of reovirus type 3 to cells that lack beta adrenergic receptor activity. AB - A previous report demonstrated both immunological crossreactivity and structural similarity between the mammalian beta adrenergic receptor and the cell surface receptor for the reovirus type 3 (14). We now demonstrate that reovirus type 3 can bind selectively and with high affinity to cells that lack beta adrenergic receptor activity (L-cells). The present study was also designed to determine what effect reovirus binding has on beta adrenergic receptor function in cells (DDT1) that possess an intact ligand binding site. Based on computer analysis of reovirus competitive inhibition curves, the apparent dissociation binding constants (Kd) for reovirus binding to DDT1 and L-cells are 0.1 nM and 0.25 nM, respectively. High affinity [125I]-iodocyanopindolol (CYP) binding to beta adrenergic receptors can also be demonstrated in DDT1 cells but not in L-cells. In agreement with these ligand binding studies, adenylate cyclase activity is stimulated by the beta receptor agonist isoproterenol in DDT1 cell membranes but not in L-cell membranes. In addition, isoproterenol increases cAMP levels in DDT1 cells but not in L-cells. Neither reovirus serotype stimulates cAMP levels in either cell line, nor do they influence beta-adrenergic agonist stimulation of cAMP in DDT1 cells. These results argue against identity of the receptors for reovirus type 3 and beta adrenergic ligands. PMID- 3027485 TI - Stress mediated changes in hypothalamic corticotropin releasing factor-like immunoreactivity. AB - Hypothalamic corticotropin releasing factor-like immunoreactivity (CRF-LI), plasma ACTH and corticosterone levels were measured by radioimmunoassay over a two hour period of restraint stress. The results of this study demonstrate a significant decrease in hypothalamic CRF-LI levels 15 and 30 minutes after the start of restraint stress which is followed by a significant increase at 60 minutes that is abolished by cycloheximide pretreatment. Plasma ACTH and corticosterone levels were significantly elevated after 15, 30, 60, 90, and 120 minutes of restraint stress. These results are consistent with a release of CRF from the hypothalamus during stress. The cycloheximide-sensitive increase in hypothalamic CRF-LI indicates that synthesis of CRF-41 occurs during prolonged stress. These results suggest that the response of an organism to exposure to a long-term, high intensity stress involves both the release and synthesis of CRF 41. PMID- 3027486 TI - Pertussis toxin reverses adenosine receptor-mediated inhibition of renin secretion in rat renal cortical slices. AB - Adenosine analogs selective for the A1 subclass of adenosine receptors, such as N6-cyclohexyladenosine (CHA), inhibit renin secretion in in vitro preparations. Ca chelation blocks the inhibitory effect, consistent with mediation by increased intracellular free Ca2+, and it has been suggested that intracellular Ca2+ could increase as a result of receptor-induced inhibition of adenylate cyclase followed by decreased Ca efflux from the renin-secreting cells. Pertussis toxin blocks receptor-induced inhibition of adenylate cyclase in many cells, and in others, it blocks receptor-induced phosphotidylinositol response. In the present studies, pertussis toxin treatment stimulated the basal renin secretory rate of rat renal cortical slices and blocked the inhibitory effect of CHA but not the inhibitory effect of K-depolarization. These data support the hypothesis that a pertussis toxin substrate, such as Ni, is involved in CHA-, but not in K-depolarization, induced inhibition of renin secretion. PMID- 3027487 TI - Cytotoxicity of commonly used nitroxide radical spin probes. AB - Since nitroxide radical spin probes are used frequently to test biophysical properties of cells, their use should be restricted to conditions that do not perturb normal cell growth and viability. Eight commonly used nitroxide radical spin probes have been tested for their effects on the survival of CHO cells. These include water-soluble spin probes Tempol, Tempamine, CTPO, CTPC and 4 maleimido-Tempo, and lipid soluble spin probes 5-Doxyl-, 12-Doxyl-, and 16 Doxylstearates. With the exception of 4-maleimido-Tempo, none of the water soluble spin labels inhibited cell survival at concentrations as high as 1 mM. At concentrations of 75 microM and higher, 4-maleimido-Tempo inhibited cell survival in a dose dependent manner. At concentrations commonly used for spin labeling of cells (30-50 microM) none of the lipid soluble spin probes tested was cytotoxic. At 100 microM only 5-Doxylstearate inhibited cell survival, whereas 12 Doxylstearate and 16-Doxylstearate had no effect. PMID- 3027488 TI - Interaction with lectin of kappa opioid binding sites solubilized from human placenta. AB - Kappa opioid binding sites from human placenta, prelabeled with 3H-etorphine and solubilized, were retained on wheat germ agglutinin (WGA) agarose and specifically eluted with N-acetylglucosamine. No significant retention was found with other immobilized lectins, including Concanavalin A (Con A), soybean seed lectin (SBA), Pisum sativum lectin (PsA), Lens culinaris Medik. lectin (LcA), and Lathyrus tingitanus lectin(LtA). About 23% of applied kappa sites were specifically eluted from WGA agarose, less than half of the proportion of rat brain opioid binding sites eluted from the same lectin (55%). Receptors from placental extracts were compared with those from other tissues enriched in either kappa or mu sites. The proportion of applied kappa sites from guinea pig cerebellum eluted specifically from WGA agarose was 36%, whereas elution of binding sites from rat thalamus and rabbit cerebellum (enriched in mu sites) was at a level of 55%. This difference in the level of retention on and elution from WGA may reflect differences in the sugar composition of the glycoproteins of the two types of receptors. Succinylation of WGA abolished its ability to retain opioid binding sites, consistent with involvement of sialic acid. However, currently available evidence suggests that differences in retention on WGA between kappa and mu sites may be due to differences in either sialic acid or N acetylglucosamine content or both. PMID- 3027489 TI - Micromolar Ca2+ requiring protease from human platelets: purification, partial characterization and effect on the cytoskeletal proteins. AB - A calcium-activated neutral protease (CANP) has been purified 2,800 fold, to near homogeneity, from human platelets. The purification procedure involved ammonium sulfate fractionation of the platelet cytosol followed by chromatography on Sephacryl S-200, DEAE-Sephacel, Agarose-Hexylamine, Agarose-Octylamine and alpha casein-Sepharose 4B affinity gel. The protease consisted of two polypeptides of Mr = 74,000 and 28,000 as judged on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. It hydrolyzed [methyl-14C] alpha-casein at a significant rate of 37 degrees C which was, therefore, used as an exogenous substrate. Microtubules and intermediate filament proteins were also susceptible to hydrolysis by the purified protease. It attained maximum activity at 0.06 uM CaCl2 and displayed two pH maxima: one at 5.5 and the other at 6.5. The protease was fully active in the presence of MnCl2 and was about 75% active with BaCl2 and SrCl2. Among the actinomycete protease inhibitors, leupeptin, antipain and pepstatin, the order of inhibition was: leupeptin greater than antipain greater than pepstatin. The protease was also inhibited by sulfhydryl modifying agents. PMID- 3027490 TI - Characterization of a serotonin transporter and an adenylate cyclase-linked serotonin receptor in the cestode Hymenolepis diminuta. AB - [3H]5-HT exhibited specific binding in membrane preparations of Hymenolepis diminuta. The specific binding was saturable, reversible and temperature dependent. A non-linear Scatchard plot was obtained in a concentration range of 11 nM - 1000 nM [3H]5-HT, which could be resolved into sites having apparent dissociation constants (KD) of 0.10 microM and 6.25 microM for the high-affinity and low-affinity components, respectively. The latter could be selectively eliminated by binding [3H]5-HT to H. diminuta membranes in the presence of 10(-3) M nitroimipramine. Drug displacement studies, using 0.20 microM and 2.0 microM [3H]5-HT, revealed that while low-affinity [3H]5-HT binding was displaced by unlabelled 5-HT and inhibitors of 5-HT uptake, high affinity [3H]5-HT binding was affected only by tryptamine derivatives and, to a lesser extent, methysergide. In addition, high-affinity binding was stimulated by MgCl2 while low-affinity binding showed sodium-dependency. The data implicate the low-affinity site as a putative 5-HT transporter and the high-affinity site as a putative 5-HT 1 receptor. Exposure of H. diminuta membranes to 5-HT resulted in a 3-4 fold stimulation of cAMP levels. The EC 50 for the 5-HT-induced activation of adenylate cyclase (0.76 microM) was of the same order of magnitude as the apparent KD for high-affinity binding. Furthermore, the order of drug potency for the elevation of cAMP levels by 5-HT agonists and reversal by 5-HT antagonists was identical to the order of drug potency for the inhibition of high-affinity binding, suggesting linkage of the putative 5-HT 1 receptor to adenylate cyclase in H. diminuta. PMID- 3027491 TI - Platelet receptors for atrial natriuretic peptide in man. AB - The aim of our study was to characterize the receptor binding of human alpha atrial natriuretic peptide (ANP) to human blood cells. Whereas no receptors were detected on red cells, on mononuclear cells and on granulocytes, we found ANP receptors on human platelets. The binding studies were performed by incubating 40 X 10(6) platelets with 125I-ANP and with the competing ligand, when used, in a total incubation volume of 1 ml. Centrifugation was used to separate bound from free hormone. Specific binding of 125I-ANP was rapid, saturable and reversible. A steady state was achieved within 90 minutes. Scatchard analysis of saturation and competition experiments demonstrated the existence of one class of high affinity binding sites for ANP with a Kd of 8-16pM and 10-26 receptors per cell. The Kd obtained in our binding studies was in the range of physiological ANP concentrations in human plasma (8-20pM). Although characterization of platelet ANP receptors has the inherent disadvantage that there are only few of them, they could be a useful model to investigate the ANP receptor-status under different physiological and pathological conditions in man. PMID- 3027493 TI - AIDS and the laboratory. PMID- 3027492 TI - Effect of GABA and benzodiazepines on testicular androgen production. AB - We have evaluated the effect of Ro5-4864, a selective probe to label peripheral type benzodiazepine receptor, on "in vitro" testicular androgen production. Decapsulated testes from adult rats showed a significant increase in the basal and hCG-stimulated testosterone secretion into the medium in response to 10(-5) M, 10(-6) M, and 10(-7) M Ro5-4864. In addition, we have studied the changes in testicular GABA content at three different ages and we found its highest concentration at 31 days of age. When we evaluated the effect of GABA on "in vitro" androgen production at different stages of gonadal maturation we observed that the highest concentration of GABA (10(-6) M) was able to modify the basal and hCG-stimulated androgen production from adult (60 days) and pubertal (45 days) testes. In addition, when prepubertal testes (31 days) were incubated under basal conditions, 10(-6) M GABA induced a significant increment of androstanediol production, while the stimulatory effect of hCG was reduced in the presence of the same GABA concentration. The present results suggest that GABA plays a physiological role in the regulation of rat testicular androgen production depending on the stage of sexual maturation. PMID- 3027494 TI - [Scintigraphy in focal lesions of the bones]. PMID- 3027495 TI - In vitro induction of human polymorphonuclear leukocyte damage by Junin virus. PMID- 3027496 TI - Comparative studies of fibrogenic properties of wollastonite, chrysotile and crocidolite. AB - Fibrogenous effects of wollastonite, chrysotile and crocidolite have been investigated in experiments on animals. (Wollastonite is a calcium silicate of fibrous structure, occasionally applied as asbestos substitute). The experimental animals were intratracheally administered single doses of 50 mg of the test material suspended in 0.6 ml of physiological NaCl solution. The animals were decapitated 3, 6 and 9 months after the experiment. Fibrogenic properties were evaluated basing on the weight of wet lungs, content of lipids and hydroxyproline in lungs, as well as histopathological tests of lungs, mediastinal lymph nodes and spleen. Histopathological preparations were evaluated using an optical, microscope or transmission electron microscope. The findings of biochemical, pathomorphological, and ultrastructural studies demonstrated slight fibrogenic effects of wollastonite, as compared to chrysotile and crocidolite. PMID- 3027497 TI - [The risk of death from malignant tumors is dependent on the amount of exposure to asbestos dust]. AB - The investigation was carried out in an asbestos plant producing yarn, cords, gaskets and frictional products. Only chrysotile asbestos was used in the production process. The follow-up covered a cohort composed of 2403 men and 1190 women employed in the plant of various asbestos products for at least 3 months during 1945-1973. The availability was 91.3% for men and 89.6% for women. Due to considerably changed working conditions in the plant, resulting from modernization carried out in 1956, the results were analysed for a subcohort employed during 1945-1955, composed of 670 men and 349 women. Men demonstrated increased standardized mortality rates (SMR) from malignant tumours of respiratory organs and thorax under increased dust concentrations. SMR = 243,2 was statistically significant only in the subcohort of men employed during 1945 1955 in a group with a mean asbestos dust concentration considerably exceeding TLVs. The risk analysis, depending on the dose, exhibited in the subcohort of men working during 1945-1955 who were affected by the highest dose of dust, increased, over 3-fold, the risk of death from lung cancer (SMR = 327.9). In the subcohort of women the risk of death from malignant tumours was higher only in the group of high dust concentration, as compared to the reference population (SMR = 210.2). A significant increase in the risk of death from cancer of the digestive organs and peritoneum as well as malignant tumours of the liver and pancrease was found in the group of high and low dust concentrations alike.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3027498 TI - Tumor promoters as probes of protein kinase C in dog thyroid cell: inhibition of the primary effects of carbamylocholine and reproduction of some distal effects. AB - The acute effects of phorbol esters, used as probes of protein kinase C activation, were studied on dog thyroid slices incubated in vitro. The derivatives used were: tetradecanoylphorbol acetate (TPA), phorbol-12,13, didecanoate (PDD), phorbol-12,13-diacetate (PDA), and phorbol dibutyrate (PDBu) and as inactive controls, phorbol itself, phorbol-12, myristate and phorbol-13, acetate, in concentrations ranging from 5.10(-8) to 5.10(-6) mol/L. The active phorbol esters had no effect on basal cyclic AMP concentrations; they inhibited cyclic AMP accumulation induced by prostaglandin E1 but not that induced by thyrotropin (TSH) 1 mU/mL and forskolin 10 mumol/L. Phorbol esters like carbamylcholine acutely stimulated iodide organification and inhibited the stimulation of hormone secretion resulting from TSH, Cholera Toxin, forskolin, and Bu2-cyclic AMP action. These metabolic effects did not require the presence of extracellular Ca++, and could not be antagonized by Ca++ depletion or manganese addition. The active phorbol esters abolished the cyclic AMP independent increased PI turnover induced by TSH 10 mU/mL or carbamylcholine (Cchol) 10(-6) mol/L but did not affect the basal incorporation of 32P into phosphatidylinositol. They reduced the 45Ca efflux from preloaded slices below basal levels and blocked the increased 45Ca release induced by TSH and Cchol. They also inhibited the increase in cyclic GMP concentrations resulting from Cchol action but not the effect of the ionophore A23187 (10(-5) mol/L) nor the basal levels of cyclic GMP.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3027499 TI - Erythrocyte thermogenesis in hyperthyroid patients: microcalorimetric investigation of sodium/potassium pump and cell metabolism. AB - Erythrocyte thermogenesis was studied by microcalorimetry in 11 patients before and after treatment for hyperthyroidism. Cell heat production rate and intracellular Na+ and K+ levels were measured in plasma suspensions of erythrocytes with and without specific inhibition of Na/K ATPase by ouabain. The ouabain induced change in the heat production rate (the Na/K pump thermal power); the erythrocyte intracellular Na+ content and the ouabain sensitive Na+ transport were used to estimate the Na/K pump function. The mean value for heat production rate was 131 +/- 4 mW/L erythrocytes before treatment, which is significantly higher than in euthyroid subjects. A significant decrease (P less than 0.01) to normal levels was recorded following therapy. This decrease, as determined in samples with ouabain, correlated to changes in serum levels of triiodothyronine, T3, (r = .74, P less than 0.01). The Na/K pump thermal power was 11 +/- 2 mW/L erythrocytes (8 +/- 2% of total heat production rate) before and 9 +/- 2 mW/L erythrocytes (8 +/- 2%) after treatment. These two values were not different from those obtained in euthyroid subjects. The erythrocyte Na+ content decreased from 9.9 +/- 2.1 to 4.9 +/- 0.5 mmol/L erythrocytes (P less than 0.001) following normalization of thyroid function. The decrease in intracellular Na+ concentration correlated to the decrease in serum T3 levels, but only when calculated from the data obtained in samples with ouabain (r = .60, P less than 0.05). The relative increase in intracellular Na+ concentrations following addition of ouabain was significantly lower (P less than 0.05) before than after treatment for hyperthyroidism, 37 +/- 10% and 61 +/- 5%, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3027501 TI - A general method for isolation of Mu d1-8(Aprlac) operon fusions in Salmonella typhimurium LT2 from Tn10 insertion strains: chlC::Mu d1-8. AB - A general method was developed for the isolation of Salmonella typhimurium LT2 Mu d1-8 (Aprlac) operon fusions in a gene displacing a Tn10 insertion. Random Mu d1 8 fusion pools were prepared to grow phage P22 lysates which transduced chlC::Tn10 to AprTets on fusaric acid-ampicillin plates. Among these AprTets potential chlC::Mu d1-8 fusions, a simple spot test identified the fusions that were closely linked to the Tn10 insertion in chlC. Out of 68 AprTets isolates 7 chlC::Mu d1-8 fusions with a nitrate-induced Lac+ phenotype were obtained. When oxrA::Tn10 was transduced into these chlC::Mu d1-8 fusions, they became Lac- even in the presence of nitrate, confirming that they were chlC::Mu d1-8 fusions. PMID- 3027500 TI - Cloning and characterization of a linear 2.3 kb mitochondrial plasmid of maize. AB - A linear 2.3 kb DNA molecule found in maize mitochondria was cloned into pUC8. A natural deletion of this plasmid, found in cmsT and some N (fertile) types of maize plants, was mapped to one end of the plasmid. A minor sequence homology to S-2, another linear mitochondrial plasmid, was detected, as well as more significant sequence homology with chloroplast and maize nuclear DNA. Hybridization to teosinte mitochondrial DNA (mtDNA) revealed the presence of part of the maize plasmid in the high molecular weight mtDNA of the maize relatives. RNA dot hybridization indicates that the plasmid is transcribed in mitochondria. The termini of the 2.3 kb linear plasmid contain inverted repeated sequences; of the first 17 nucleotides of the termini, 16 are identical to the terminal inverted repeats of the linear S plasmids found in the mitochondria of cmsS maize plants. PMID- 3027502 TI - Different efficiency of UmuDC and MucAB proteins in UV light induced mutagenesis in Escherichia coli. AB - Two multicopy plasmids carrying either the umuDC or the mucAB operon were used to compare the efficiency of UmuDC and MucAB proteins in UV mutagenesis of Escherichia coli K12. It was found that in recA+ uvr+ bacteria, plasmid pIC80, mucAB+ mediated UV mutagenesis more efficiently than did plasmid pSE117, umuDC+. A similar result was obtained in lexA41 (Def) cells, excluding the possibility that this was due to a differential regulation by LexA of the umuDC and mucAB operons. We conclude that some structural characteristic of the UmuDC and MucAB proteins determines their different efficiency in UV mutagenesis. This characteristic could be also responsible for the observation that in the recA430 mutant, pIC80 but no pSE117 can mediate UV mutagenesis. In the recA142 mutant, pIC80 also promoted UV mutagenesis more efficiently than pSE117. In this mutant, the recombination proficiency, the protease activity toward LexA and the mutation frequency were increased by the presence of adenine in the medium. In recA+ uvrB5 bacteria, plasmid pSE117, umuDC caused both an increase in UV sensitivity as well as a reduction in the mutation frequency. These negative effects resulting from the overproduction of UmuDC proteins were higher in recA142 uvrB5 than in recA+ uvrB5 cells. In contrast, overproduction of MucAB protein in excision-deficient bacteria containing pIC80 led to a large increase in the mutation frequency. We suggest that the functional differences between UmuDC and MucAB proteins might be due to their different dependence on the direct role of RecA protease in UV mutagenesis. PMID- 3027503 TI - Nucleotide sequence of the nifLA operon of Klebsiella oxytoca NG13 and characterization of the gene products. AB - The complete nucleotide sequence of the regulatory operon nifLA of a nitrogen fixer Klebsiella oxytoca NG13 was determined, and the transcriptional start point was assigned by S1 mapping. The nifL protein (a repressor) was coded by an open reading frame of 1,485 bases, corresponding to a protein of 495 amino acids with a calculated molecular weight of 55,242. The open reading frame (1,572 bases) of the nifA protein (an activator), corresponding to a molecular weight of 58,649, was confirmed by in vitro transcription-translation experiments, using the wild type and artificially deleted nifA genes. The initiation codon (ATG) of nifA overlapped the termination codon(TGA) of nifL, sharing the two bases T and G. A conserved DNA contact point [Gln-(X)3-Ala-(X)3-Gly-(X)5-Val] common in many DNA binding proteins was found in the C-terminal region of the nifA sequence. The promoter sequences of nifLA, nifB and nifF in K. oxytoca coincided exactly with those of K. pneumoniae in the consensus regions at -12 and -26, although the overall homology in the promoter regions was 96%. Changes of four amino acids were found between the nifA coding sequences of K. oxytoca and K. pneumoniae. PMID- 3027504 TI - Cloning and complete nucleotide sequence of the Escherichia coli glutamine permease operon (glnHPQ). AB - The glutamine permease operon encoding the high-affinity transport system of glutamine in Escherichia coli could be cloned in one of the mini F plasmids, but not in pBR322 or pACYC184, by selection for restoration of the Gln+ phenotype, the ability to utilize glutamine as a sole carbon source. We determined the nucleotide sequence of the glutamine permease operon, which contains the structural gene of the periplasmic glutamine-binding protein (glnH), and indispensable component of the permease activity. The N-terminal amino acid sequence and the overall amino acid composition of the purified glutamine-binding protein were in good agreement with those predicted from the nucleotide sequence, if the N-terminal 22 amino acid residues were discounted. The latter comprised two Lys residues (nos. 2 and 6) followed by 16 hydrophobic amino acid residues and was assumed to be a signal peptide for transport into the periplasmic space. There were two additional reading frames (glnP and glnQ) downstream of glnH sharing a common promoter. It was concluded that the glnP and glnQ proteins as well as the glnH protein are essential for glutamine permease activity. PMID- 3027505 TI - Transposable multiple antibiotic resistance in Streptococcus pneumoniae. AB - A mobile genetic element, designated Tn1545, was detected in the chromosome of Streptococcus pneumoniae BM4200, a clinical isolate multiply resistant to antibiotics. The 25.3 kb element conferred resistance to kanamycin and structurally related aminoglycosides by synthesis of a 3'-aminoglycoside phosphotransferase type III (aphA-3), to macrolide-lincosamide-streptogramin B type antibiotics (ermAM), and to tetracycline (tetM). Tn1545 was self transferable to a recombination deficient S. faecalis strain where it was able to transpose to various sites, induce insertional mutations and was apparently cleanly excised. The element also conjugated to and transposed to the chromosome of S. faecalis, S. lactis, S. diacetylactis, S. cremoris, S. sanguis, Staphylococcus aureus, and Listeria monocytogenes. The properties of the conjugative transposon Tn1545 could account for the sudden emergence, rapid dissemination, and stabilisation of multiple resistance to antibiotics in S. pneumoniae in the absence of plasmids. PMID- 3027506 TI - The recQ gene of Escherichia coli K12: primary structure and evidence for SOS regulation. AB - A 2,695 bp chromosomal segment of Escherichia coli K12 containing the recQ gene was sequenced. Analysis of the sequence revealed an open reading frame thought to represent recQ, with a clockwise direction of transcription relative to the standard genetic map of E. coli K12 and having a coding capacity for a protein of Mr 68,350. The -10 region of the presumptive recQ promoter overlapped the putative terminator for the upstream gene pldA, and was immediately followed by a 15 bp stretch of DNA bearing a strong resemblance to the reported sequences of LexA repressor binding sites. This latter finding suggested the possibility of SOS regulation of recQ gene expression, which was substantiated by experiments with recQ-lacZ fusions. PMID- 3027508 TI - Molecular cloning and characterization of ARO7-OSM2, a single yeast gene necessary for chorismate mutase activity and growth in hypertonic medium. AB - The chorismate mutase structural gene, ARO7, which is necessary for both phenylalanine and tyrosine biosynthesis was cloned by complementation in yeast. Genetic analysis showed that ARO7 was identical to a gene necessary for growth in hypertonic medium, OSM2, which mapped nearby. After restriction mapping and subcloning of the plasmid, the cloned gene was used to detect mRNA levels in several growth conditions. Enzyme activities were measured in various genotypes. At our level of detection ARO7-OSM2 is a low level constitutively expressed gene. PMID- 3027507 TI - Mutations in the right boundary of Saccharomyces cerevisiae centromere 6 lead to nonfunctional or partially functional centromeres. AB - Centromeres most likely consist of DNA (CEN DNA) interacting with specific proteins. In Saccharomyces cerevisiae a clear picture has emerged of a 120 bp sequence that is characteristic of CEN DNA. We have investigated the 25 bp centromere DNA element (CDEIII) that represents the right part of a CEN DNA. We showed using a series of mutants generated in vitro that the right most triple A of the consensus sequence TGT.T.TG.. TTCCGAA.....AAA participates in the assembly of a functional centromere and that no further sequences to the right are needed. Distance changes between the centre dyad TTCCGAA and the triple A have two effects: Addition of one base pair leads to a reduction, and addition of two or four base pairs to a loss of centromere function implying a participation of the centre dyad and the triple A region in protein binding. Indeed, a synthetic oligonucleotide of 39 bp containing CDEIII shows specific protein binding. PMID- 3027511 TI - Revised sequence of the nusA gene of Escherichia coli and identification of nusA11 (ts) and nusA1 mutations which cause changes in a hydrophobic amino acid cluster. AB - The mutant nusA DNAs (nusA11 and nusA1) were sequenced. Single base substitutions caused by these mutations were found in the coding region of nusA. The nusA11 mutation, which is conditionally lethal, substituted Thr for the 181st Ala. Also, nusA1, which restricts lambda growth, substituted Ala for the 183rd Ser. These two positions were located in the same hydrophobic amino acid cluster. This cluster seemed to be an essential region in the functional domain of NusA. In the course of these experiments, several mistakes in the published nusA nucleotide sequence were found. These errors are revised in this article. The molecular weight of NusA is accordingly revised to 54,430. PMID- 3027509 TI - Mutagenesis by random linker insertion into the lamB gene of Escherichia coli K12. AB - Gene lamB encodes an outer membrane protein involved in maltose and maltodextrin transport as well as phage adsorption. The active form is a trimer. We characterized 11 mutations in lamB, obtained after random insertion of a BamH1 linker and screening for stable immunodetectable mutant proteins. Six mutations resulted in the loss of the distal part of the LamB protein either by deletion (five cases) or frameshift (one case). The six corresponding proteins had all lost the ability to confer phage sensitivity and the capacity to grow on dextrins, and to yield immunodetectable oligomers. Induction of a high level of the four longest of these proteins was toxic to the cell. Five other mutations were due to in-frame insertions. In four cases, the corresponding proteins still had the ability to yield immunodetectable oligomers, to confer phage sensitivity and the capacity to grow on dextrins and were not toxic on induction. In one case (AJC73), the mutant protein had lost the first three properties and was toxic on induction. Deletions and duplications between some of the inserts were also constructed and studied. To account for our results we present a hypothetical scheme in which trimerization would not only be needed for phage sensitivity and growth on dextrins but also for proper insertion into the outer membrane. The C terminus of the protein, as well as other regions such as the site of mutation AJC73, would be required for the formation of stable trimers. We tentatively interpret toxicity as due to improper insertion into the outer membrane. Our results also show that it is possible to insert several amino acids (up to 11 in one case) at a number of positions in LamB without appreciably affecting its export and activities. PMID- 3027510 TI - Regulation of expression of the gene for vitamin B12 receptor cloned on a multicopy plasmid in Escherichia coli. AB - The btuB gene of Escherichia coli codes for a protein (BtuB) located in the outer membrane. BtuB is the receptor for vitamin B12 (cyanocobalamin). We have cloned the btuB gene into pUC8 using transposon Tn5 as the marker to first isolate several parts of the relevant DNA fragment from the specialized transducing phage lambda darg13. After reconstitution of the gene, Tn5 was removed by selecting for spontaneous excision. The partial nucleotide sequence and transcriptional start of the btuB gene were determined. The BtuB+ plasmid allowed a large amplification of the synthesis of BtuB, resulting in a 65-fold increased level of vitamin B12 binding. The level of vitamin B12 binding was reduced by a factor of 22 when cells were grown in the presence of high concentrations of vitamin B12. The regulation of the gene was studied in more detail by the use of a protein fusion between the extreme amino-terminus of BtuB and beta-galactosidase of E. coli. The kinetics of repression and derepression were consistent with the presence in the cells of a large amount of a regulatory molecule exhibiting an apparent Km for vitamin B12 of 3 microM. PMID- 3027513 TI - Evaluation of the efficacy of split-product trivalent A(H1N1), A(H3N2), and B influenza vaccines: protective efficacy. AB - A total of 1,995 primary school children (1,464 vaccinees and 531 non-vaccinees) were studied to evaluate the protective efficacy of Tween-ether split trivalent A(H1N1), A(H3N2), and B influenza vaccines by comparison of the incidence of confirmed infection in two groups during 1980 to 1984. During the study period, epidemics caused by antigenically different influenza viruses, that is A(H1N1) epidemics in 1981 and 1984, a B epidemic in 1982 and an A(H3N2) epidemic in 1983, were experienced, and at the same time strains changed by antigenic drift were frequently isolated. In these epidemics, 61% to 87% of the children reported respiratory illnesses and 18% to 48% of the illnesses were influenza confirmed by seroconversion. Throughout these four epidemics, the incidence of confirmed infection among the vaccinees (7.8% to 33.8%) was 6.5% to 34.8% lower than that among the nonvaccinees (35.4% to 51.6%), demonstrating that the vaccine was effective (X2 = 76.34, P less than 0.001). However, this effectiveness was not seen in an epidemic in one of the entrant schools in 1984, possibly caused by a strain with intense antigenic drift. On the basis of data on incidence of various symptoms, duration of fever and the number of days of absence from class, it was considered that clinical symptoms in the vaccinees were milder than those in the nonvaccinees. When the titers of hemagglutination-inhibiting (HAI) antibody against the vaccine strains were measured, the protective level of HAI antibody giving less than or equal to 50% incidence of infection was 1:64, but it increased to 1:256 in the 1984 epidemic, reflecting the high rate of isolates with intense antigenic drift. PMID- 3027514 TI - Enhanced interleukin 1 production by alveolar macrophages and increase in Ia positive lung cells in silica-exposed rats. AB - Inhalation exposure to silica dust enhanced interleukin 1 (IL-1) production by alveolar macrophages (AM), which is attributable to an increase in Ia-positive lung cells. While the proportion of Ia-positive cells in lavaged bronchoalveolar cells (BAC) was much lower (0-3%) in unexposed control rats, about a third of the rats that inhaled silica showed higher proportions (8.0-18.5%); these were designated "Ia-high" exposed animals. The number of total cells, Ia-positive cells and lymphocytes in BAC was significantly increased (P less than 0.05, P less than 0.001, and P less than 0.001, respectively) in these "Ia-high" exposed animals, compared to the control animals. Adherent AM populations obtained from BAC preparations also contained significantly higher (P less than 0.001) proportions of Ia-positive cells in the "Ia-high" exposed animals. When these adherent AM cultures were stimulated with lipopolysaccharide, IL-1 activity of the culture supernatants was enhanced and was significantly higher (P less than 0.001) in the "Ia-high" exposed rats, compared to the control animals. These results indicate that silica-exposure can induce populational changes in lung cells and also activation of AM associated with the increase in Ia-positive cells. PMID- 3027515 TI - Effects of disodium ethane-1-hydroxy-1,1-diphosphonate (EHDP) on interleukin 1 production by macrophages. AB - The effects of disodium ethane-1-hydroxy-1,1-diphosphonate (EHDP) on the in vitro functions of guinea pig macrophages were studied. A high dose (1 mg/ml) of EHDP inhibited interleukin 1 (IL 1) production by oil-induced peritoneal macrophages stimulated with muramyl dipeptide (MDP), lipopolysaccharide (LPS), phorbol myristic acetate (PMA), heat-aggregated IgG2 or calcium ionophore A23187. On the other hand, low doses (less than 0.125 mg/ml) of EHDP augmented the MDP induced IL 1 production by macrophages. This biphasic effect was also observed when macrophages were exposed to EHDP at 37 C for 24 hr and then stimulated with IL 1 inducers. Superoxide anion generation induced by formyl peptide or PMA was not affected by preincubation of the macrophages with doses of EHDP up to 1 mg/ml. Adherence and spreading of macrophages was inhibited by EHDP in a dose dependent manner without affecting cell viability. These results demonstrated that EHDP acted on macrophages directly and modulated IL 1 production in vitro. PMID- 3027512 TI - Conjugal acquisition and stable maintenance of Ent plasmids in nontoxigenic wild type strains of Escherichia coli. AB - In spite of the ability of the genetic determinants for enterotoxin production to be conjugally transferred, mobilized or transposed, enterotoxigenic Escherichia coli (ETEC) isolated from diarrheal patients is restricted to certain serotypes. Four conjugative enterotoxigenic plasmids (Ent plasmids) encoding either a heat labile enterotoxin or a heat-stable enterotoxin or both and belonging to one of three incompatibility groups IncFI, IncHl, or IncX, were examined for their transferability to and stability in 157 nonenterotoxigenic Escherichia coli strains belonging to various serotypes and 89 clinical isolates nonenterotoxigenic but belonging to those serotypes in which ETEC from diarrheal patients are usually found. The serotypes of the strains to which Ent plasmids were efficiently transferred and in which they were maintained stably were not always the serotypes in which ETEC had usually been found and vice versa. The frequencies of transfer of four Ent and two R plasmids to each of the 157 recipients were correlated with each other, indicating that the frequency of transfer of the plasmid is not determined by a resident plasmid, if there is one, but by a recipient factor which commonly affects transferability to all donors. These results have led to the conclusion that the reason why only certain serotypes are found among ETEC isolated from diarrheal patients is not the ability of these strains specifically and preferentially to acquire and maintain the Ent plasmids. PMID- 3027516 TI - Phantom material for quantitative evaluation of MR images. AB - Reference material for quantitative Magnetic Resonance Imaging (MRI) has been developed by combining heavy and light water (D2O, H2O), gelling agent agar, and paramagnetic transition metal ions (Gd3+). The process of preparation is described. By in vitro measurement of relaxation times, T1 and T2, it can be shown that tissue relevant values are achievable. The influence of different relaxation times, as well as the effect of different proton spin densities, on MRI signal intensities is determined. Some similarities existing between the magnetic resonance behaviour of tissue water and phantom substance water are discussed. PMID- 3027517 TI - Isolated rectal gland cells: oxygen consumption and hormonal stimulation. AB - Cells isolated from rectal glands of Squalus acanthias, using collagenase and hyaluronidase digestion, retained normal morphological characteristics as judged by light microscopy of 1-micron plastic sections. Their oxygen consumption per unit weight was comparable to that of intact rectal gland studied either in situ, or by isolated perfusion, as well as that of rectal gland slices. Cellular respiration was stimulated by dibutyryl cyclic AMP and theophylline or by vasoactive intestinal peptide which stimulate secretion of chloride by the intact gland. Stimulated oxygen consumption was inhibited by ouabain and bumetanide and was proportional to the concentration of sodium or chloride in the incubation solution. The oxygen consumption of these cells parallels the secretory and metabolic behavior of the intact rectal gland, suggesting that it reflects energy demands for ion transport. The relative ease with which a homogeneous preparation of viable and active cells can be obtained and the apparent preservation of many of their key functional characteristics make this preparation a useful tool for the study of hormone-stimulated ion transport. PMID- 3027518 TI - Jejunal phosphate transport is not regulated by the PTH-adenylate cyclase system. Further studies on the contrasting features between intestinal and renal phosphate transport mechanisms. AB - Direct application of parathyroid hormone (PTH) to renal proximal tubule in vitro stimulates the adenylate cyclase system which in turn causes reduction in phosphate (P) reabsorption. Similar application of parathyroid extract to rat jejunum fails to induce changes in P transport. In the present study we examined the basis for this PTH-unresponsiveness in rat jejunum. The effect of PTH, and another peptide hormone, salmon calcitonin (SCT) and sodium fluoride (NaF) on jejunal adenylate cyclase activity was first examined in both vitamin D-deficient (-D) rats and similar rats after 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] repletion. Jejunal cyclic 3',5'-AMP (cAMP, pmol/mg protein/min) increased from 11.0 +/- 1.1 in -D rats to 23.0 +/- 2.2 in 1,25(OH)2D3-repleted rats (p less than 0.001). Neither PTH nor SCT had any effect on adenylate cyclase activity in jejunum from either -D or 1,25(OH)2D3-repleted rats. NaF caused the anticipated stimulation in cAMP generation, a response independent of the vitamin D nutritional status of the animal [-D: 76.8 +/- 1,25(OH)2D3: 86.9 +/- 8.5]. We then examined the question if exogenous cAMP, which reduces renal P reabsorption, also causes decrease in intestinal P absorption. The effect of dibutyryl cAMP (DbcAMP) on transepithelial, net P absorption was examined in short-circuited jejunal epithelia from rats maximally stimulated by 1,25(OH)2D3. DbcAMP caused the anticipated increase in short-circuit current (+212%) without affecting the net, active P absorption. We conclude that, unlike the renal proximal tubule, the adenylate cyclase system in jejunum is insensitive to PTH, and the P absorptive mechanism is resistant to cAMP. PMID- 3027519 TI - High Ca2+ inhibits peptide hormone-dependent cAMP production specifically in thick ascending limbs of Henle. AB - We showed previously that increasing Ca2+ concentration in the incubation medium suppressed cAMP production in response to vasopressin (AVP), glucagon or forskolin in the medullary thick ascending limb of Henle (MTAL) but not in medullary collecting tubules of mouse kidney. In the present study, we examined, using nephron segments dissected from mouse kidney, whether the inhibitory effect of high Ca2+ is specific to MTAL. Increasing Ca2+ in the incubation medium from 1 to 5 mM inhibited cAMP production in response to parathyroid hormone (PTH), calcitonin, AVP or glucagon in cortical thick ascending limbs of Henle (CTAL), but dit not inhibit cAMP production stimulated by PTH or calcitonin in proximal convoluted tubules and that by AVP in cortical collecting tubules. In CTAL, high ambient Ca2+ also inhibited cAMP production stimulated by forskolin. Thus, our present data show that high Ca2+ inhibits cAMP production specifically in thick ascending limbs of Henle but not in the other nephron segments. High ambients Ca2+ may inhibit adenylate cyclase at postreceptor site(s) one of which may be the catalytic unit of the enzyme in TAL. PMID- 3027521 TI - How do the elderly fare in English society? Care of the aged in Oxford, England. (Part 2). PMID- 3027520 TI - Nurse recruiters report nursing shortage is back: many reactivating programs to attract nurses. PMID- 3027522 TI - Evaluation of models for analysis of radioligand binding data. AB - In the presence of agonists, many neurotransmitter receptors interact with regulatory components, resulting in the formation of a ternary complex composed of agonist, receptor, and a regulatory component, and in biphasic or shallow dose response curves for inhibition of the binding of a radiolabeled antagonist by agonists. Complex dose response curves are often analyzed using equations that describe a model that assumes the presence of two independent populations of receptors (two-independent-receptor model) or equations that describe a model that assumes the reversible interaction of agonist-occupied receptors with a regulatory component (ternary complex model). In this study, the ability of these models to provide good estimates of the concentration of the regulatory component and of the affinities involved in formation of the ternary complex was evaluated. Dose response curves were generated by a computer using an equation that describes the ternary complex model. Analysis of the dose response curves with the two-independent-receptor model resulted in good estimates of the concentration of the regulatory component and of the affinity of the receptor for the agonist. Reliable estimates of the other affinity constants that relate to formation of the ternary complex could not be obtained. Analysis of the dose response curves with the ternary complex model resulted in good estimates of the concentration of the regulatory component and of the affinity constants that relate to formation of the ternary complex. Random error was added to the data points that made up the dose response curves to simulate error observed in experiments with biological systems. Analysis of the dose response curves that contained random error with the two-independent-receptor model yielded results similar to those obtained from analysis of dose response curves that did not contain random error. Analysis of the dose response curves that contained random error with the ternary complex model resulted in good estimates of the concentration of the regulatory component and of the affinity of the receptor for the agonist. However, reliable estimates of the other affinity constants involved in formation of the ternary complex could not be obtained. Thus, caution should be exercised when interpreting results of the analysis of dose response curves with either the two-independent-receptor model or the ternary complex model. PMID- 3027523 TI - Alpha 1-adrenergic receptor-linked guanine nucleotide-binding protein in muscle and kidney epithelial cells. AB - We have studied the interaction of guanine nucleotides with alpha 1-adrenergic receptors of two cloned cell lines, the Madin Darby canine kidney (MDCK-D1) cells and BC3H-1 muscle cells. Although guanylylimidodiphosphate, Gpp(NH)p, had no effect on the affinity or the total number of [3H]prazosin-binding sites in membranes prepared from these cells, the nucleotide decreased the apparent affinity of the agonists (-)-epinephrine and (-)-norepinephrine in competing for [3H]prazosin-binding sites in both cell types. A maximal effect of Gpp(NH)p occurred at 10 microM. Guanine nucleotides were significantly more effective in shifting agonist affinity for the alpha 1 receptor than adenine nucleotides, and Mg2+ was required to observe a maximal effect. Binding of agonist to alpha 1 adrenergic receptors activated phosphatidylinositol (PI) hydrolysis in both cell types but had no effect on membrane adenylate cyclase activity. Incubation of MDCK cells for 19 hr with 100 ng/ml pertussis toxin, which eliminated the ability of pertussis toxin added to membranes to ADP-ribosylate 39-41-KDa substrate(s), failed to alter binding of agonists to alpha 1-adrenergic receptors, the ability of Gpp(NH)p to regulate agonist binding to these receptors, or epinephrine stimulated PI hydrolysis and prostaglandin E2 production. Incubation of BC3H1 cells with pertussis toxin had no effect on the ability of epinephrine to stimulate PI turnover. These results show that binding of agonists to alpha 1 adrenergic receptors in mammalian kidney and muscle cells is regulated by guanine nucleotides, presumably by interaction with a guanine nucleotide-binding (G) protein. The failure of the G-protein to regulate adenylate cyclase activity and the lack of effect of pertussis toxin to alter receptor-mediated binding or functional activity suggests that a G-protein other than Gs, Gi, or Go interacts with alpha 1-adrenergic receptors in kidney and smooth muscle. PMID- 3027524 TI - Leukotriene-induced hydrolysis of inositol lipids in guinea pig lung: mechanism of signal transduction for leukotriene-D4 receptors. AB - Addition of leukotriene D4 (LTD4) to [3H]myo-inositol-labeled guinea pig lung induced rapid breakdown of inositol lipids. Formation of [3H]inositol trisphosphate was rapid, with a peak of 140-160% of the control level, 30 sec post-treatment. Formation of [3H]inositol bisphosphate and [3H]inositol monophosphate ([3H]IP1) was also rapid in the presence of LiCl. LTD4-induced [3H]IP1 formation was concentration dependent, stereoselective, and not inhibited by the cyclooxygenase inhibitor, indomethacin. Agonist analogs of LTD4 and leukotriene E4 also induced dose-dependent increases in the synthesis of [3H]IP1. The rank order potency of the agonist-induced [3H]IP1 formation was equivalent to those reported for LTD4 receptor binding, smooth muscle contraction, and thromboxane B2 biosynthesis. Furthermore, a specific receptor antagonist, SKF 102922, inhibited LTD4-induced [3H]IP1 formation in guinea pig lung. These studies suggest that LTD4 may interact with membrane receptor and activate a phospholipase C, which in turn induces the hydrolysis of inositol lipids. The hydrolysis products, diacylglycerol and inositol trisphosphate, can be regarded as the intracellular messengers for LTD4 receptors in guinea pig lung. This concept may explain a variety of pharmacological effects of leukotrienes in different types of target cells or tissues. PMID- 3027525 TI - Photoaffinity labeling of peripheral-type benzodiazepine-binding sites. AB - The use of a novel photoaffinity label for the peripheral-type benzodiazepine binding site is described. This compound, PK 14105, has high affinity (4 nM) and selectivity for cardiac benzodiazepine-binding sites. Under ultraviolet light, PK 14105 couples covalently to an 18,000-Da membrane protein which apparently corresponds to the (or a part of the) cardiac benzodiazepine-binding site. Since covalent attachment of PK 14105 totally precludes the binding of other ligands to this binding site, it is suggested that, during ultraviolet irradiation, this compound inserts covalently into the binding domain of the peripheral-type benzodiazepine-binding site. PMID- 3027526 TI - Molecular structure of 3-(methoxycarbonyl) amino-beta-carboline, a selective antagonist of the sedative effects of diazepam. AB - The X-ray crystal structure of 3-(methoxycarbonyl) amino-beta-carboline, a selective antagonist of the sedative effects of diazepam having a high affinity for the benzodiazepine receptor, has been determined. The results were compared with structural information obtained from this compound, both in the solid state and in dilute solution, by use of Fourier transform infrared spectroscopy. Its X ray structure was also compared with those of two other active beta-carbolines, methyl beta-carboline-3-carboxylate and N-ethyl-3-carbamoyl-beta-carboline. The crystal packing characteristics of 3-(methoxycarbonyl) amino-beta-carboline differ from those of these two beta-carbolines in both the pattern of intermolecular hydrogen bonding and the quality of their pi-pi stacking interactions. The relevance this may have to the selective activity of 3 (methoxycarbonyl) amino-beta-carboline is discussed. PMID- 3027527 TI - Dual regulation of beta-melanotropin receptor function and adenylate cyclase by calcium and guanosine nucleotides in the M2R melanoma cell line. AB - Binding of beta-melanotropin (beta-MSH) and subsequent activation of adenylate cyclase in the M2R mouse melanoma cell line is strongly dependent on the concentration of extracellular free calcium. This effect can be demonstrated both in the intact cell and in a plasma membrane preparation derived therefrom, using an EGTA buffer system. In contrast, stimulation of adenylate cyclase by prostaglandin E1, forskolin, or guanosine 5'-O-(2-thiotriphosphate) is calcium insensitive. It is shown that calcium increases the binding affinity of beta-MSH for its receptor by a factor of 20 (from 400 nM to 20 nM) without affecting maximal hormone binding. At supersaturating concentrations of beta-MSH (greater than 200 nM) binding gradually becomes calcium independent. Hormone-receptor complexes formed in the presence of calcium dissociated rapidly (less than or equal to 2 min) and reversibly upon the elimination of calcium by excess EGTA. Among nine divalent metal cations tested, calcium was found to be the most effective in facilitating hormone binding. Whereas calcium promotes beta-MSH binding, GTP and its stable analogs lead to a reduction in both maximal binding (65%) and affinity (2-fold). These effects are calcium independent, suggesting that the reciprocal control of beta-MSH binding by calcium and guanosine nucleotides is mediated by two separate and independent mechanisms. PMID- 3027528 TI - GTPase and adenylate cyclase desensitize at different rates in NG108-15 cells. AB - The time course of opioid receptor binding disappearance and loss of responsiveness of the opioid-controlled GTPase and adenylate cyclase were compared in membranes derived from NG108-15 cells pretreated with the opioid peptide agonist [D-Ala2,D-Leu5]enkephalin (DADLE). Upon pretreatment with DADLE, a rapid desensitization of the opioid-stimulated GTPase occurred with a time course distinguishable as two exponential components having respective half-lives of 5-9 and 60-80 min. Opioid receptor binding activity, as assessed using [3H]diprenorphine, also decayed as two exponential components whose half-lives were similar to those for GTPase desensitization (7 and 120 min). However, when [3H]diprenorphine binding was measured in the presence of sodium and GTP, only the second, slow component was apparent. In contrast, desensitization of the opioid-controlled adenylate cyclase occurred as only one exponential decaying process, displaying a half-life of 57 min. Whereas the loss of responsiveness of GTPase to DADLE was entirely accounted for by a reduction in the maximal stimulation produced acutely by DADLE, desensitization of adenylate cyclase was characterized by both a decrease in maximal inhibition and a shift to the right of the EC50 of the agonist in inhibiting acutely the enzyme. In addition, after 1 hr of pretreatment with DADLE, the opioid-stimulated GTPase was desensitized by 65%, whereas 80% of maximal inhibition of adenylate cyclase could still be achieved. We suggest that: the rapid loss of responsiveness of the opioid-GTPase system results from an uncoupling between the receptor and the nucleotide-binding regulatory protein (N); the fast decaying GTPase activity appears to be not directly related to the opioid-mediated inhibition of adenylate cyclase; and the slow decaying GTPase activity, as well as the desensitization of the opioid adenylate cyclase, is most likely accounted for by down-regulation of the opioid receptor. These findings may indicate that part of the opioid-stimulated GTPase in the membrane is not involved in inhibition of the cyclase and could reflect the activity of a regulatory protein which couples opioid receptors to another membrane effector. Alternatively, they might be interpreted on the basis of a model which involves a tight coupling between receptor activation and N protein and a large amplification mechanism between N protein and adenylate cyclase. PMID- 3027529 TI - Phorbol ester-mediated inhibition of vasopressin and beta-adrenergic responses in a vascular smooth muscle cell line. AB - We have reported previously that in the vascular smooth muscle cell line A-10 (ATCC CRL 1476), vasopressin stimulated phosphatidylinositol turnover Ca2+ efflux and inhibited isoproterenol-stimulated cAMP accumulation. Here we report that pretreatment of these cells with phorbol dibutyrate, an activator of protein kinase C, attenuated the responses to vasopressin and isoproterenol. This effect was concentration dependent and could be observed after pretreatment for 2 min. 4 alpha Phorbol 12,13-didecanoate, which does not activate protein kinase C, did not attenuate the responses. These data suggest that activation of protein kinase C by phorbol dibutyrate attenuates the responses of vascular smooth muscle cells to isoproterenol and vasopressin. Although phorbol ester did not affect [3H]-8 arginine vasopressin binding to intact cells, it appeared to uncouple vasopressin receptors from guanine nucleotide-binding protein. PMID- 3027530 TI - In vivo effects of mercury (II) on deoxyuridine triphosphate nucleotidohydrolase, DNA polymerase (alpha, beta), and uracil-DNA glycosylase activities in cultured human cells: relationship to DNA damage, DNA repair, and cytotoxicity. AB - The effect of mercuric acetate on the activities of deoxyuridine triphosphate nucleotidohydrolase (dUTPase), DNA polymerase (alpha, beta), and uracil-DNA glycosylase has been studied in cultured human KB cells. There was a dose- and time-dependent inactivation of both dUTPase and DNA polymerase alpha activities by mercuric acetate. In cells exposed to low concentrations (10 microM) of mercuric acetate, dUTPase was most sensitive to inhibition with 30% of the activity being inhibited after a 1-hr exposure. At higher concentrations or for longer exposure times, DNA polymerase alpha was most sensitive to inhibition with greater than 60% of the activity being inhibited by 25 microM mercuric acetate after a 15-min exposure. There was no inhibition of DNA polymerase beta or uracil DNA glycosylase activities in cells exposed to 50 microM mercuric acetate for 90 min. In fact, there was a time- and dose-dependent activation of uracil-DNA glycosylase activity with maximum activation occurring in cells exposed to 50 microM mercuric acetate. The inhibition of dUTPase and DNA polymerase alpha activities and the activation of uracil-DNA glycosylase activity correlated with the induction of single-strand breaks in DNA by mercuric acetate and with the decrease in cell viability. PMID- 3027531 TI - Effects of glucagon and insulin on the cyclic AMP binding capacity of hepatocyte cyclic AMP-dependent protein kinase. AB - Extracts obtained from rat hepatocytes incubated with saline, glucagon or insulin were electrophoresed on polyacrylamide gels and then assayed for cyclic (3H)AMP binding capacity. Analysis of the binding patterns demonstrated that glucagon dissociated a holoenzyme of cyclic AMP-dependent protein kinase in a dose dependent manner. The increase in free regulatory subunits and, hence, in free catalytic subunits explains the activation of this enzyme by glucagon in the liver. Insulin decreased both the amount of cyclic (3H)AMP bound to the holoenzyme and the capacity of the enzyme to be dissociated when the extracts were incubated with increasing concentrations of this cyclic nucleotide. We propose that these insulin-induced effects are determined by an inhibition of the cyclic AMP binding capacity of this protein kinase. This mechanism could account for the inactivation of cyclic AMP-dependent protein kinase that insulin causes in the liver. PMID- 3027532 TI - Developmental changes in the chromatin of the brain of the rat: analysis by nick translation. AB - Nick translation of nuclei of the brain of 3- 14- and 30-day old rats was carried out following their digestion by DNase I. The incorporation of 3H-dTMP at 14- and 30-day is significantly lower than at 3-day. This may be due to a lower proportion of active chromatin (DNase I hypersensitive sites) and condensation of chromatin with progressive development. When nuclei were digested by EcoRI and then nick-translated, the incorporation of 3H-dTMP showed the same pattern. Since the EcoRI sites are believed to be randomly distributed, the overall conformation of chromatin including the DNase I sensitive sites seems to undergo increasing compaction with development. PMID- 3027533 TI - [Human T-lymphotrophic retroviruses]. AB - Investigations of retroviruses, possible etiological agents of T-cell human leukemias and acquired immunodeficiency syndrome are reviewed. Main attention is paid to the genome structure and function. PMID- 3027534 TI - [Detection and preliminary characteristics of the transforming gene (oncogene) of Ha-ras type in human stomach cancer]. AB - High molecular weight DNA prepared from three undifferentiated human stomach carcinomas was assayed for transforming activity by transfection of mouse NIH 3T3 cells. One tumor DNA sample (stomach carcinoma CaVSt) induced (the transfection efficiency: 0.02 transformants/micrograms DNA X 10(-6) cells) transformation of NIH 3T3 recipient cells. Transforming gene of Ha-ras type was identified in transformants derived from this human carcinoma. The genetic lesion responsible for the activation of the CaVSt Ha-ras oncogene is not localized in the 12-th codon for p21c-Ha-ras protein. PMID- 3027535 TI - [Effect of ligand orientation on the redox potential of homologous proteins]. AB - Using experimental g-values of homologous cytochromes isolated from the horse heart and bacterial cyt-c-551 from Pseudomonas aeruginosa, the electron energy difference of redox-orbital of Fe(III) ion of heme was calculated during reduction. Data are in a good accordance with experimental redox-potential values for these proteins. The model gives opportunity to predict redox-potential values for other homologous proteins. PMID- 3027536 TI - [Netropsin, distamycin A, bis-netropsins as selective inhibitors of restrictases and DNAse I]. AB - The simultaneous analysis of DNAase I "footprinting" data and restriction endonucleases inhibition data was performed on the same DNA end-labelled fragment. The inhibition induced by netropsin, a number of bis-netropsins and distamycin A was investigated. These experiments led us to the following conclusions. The restriction endonucleases inhibition by the ligands is caused by the ligand molecules binding in the close vicinity to the restriction endonuclease recognition sequence. The zone of +/- 4 bp from the center of the restriction endonuclease recognition sequence can be defined as the zone of the influence of the bounded ligand on the restriction endonuclease. But in this case the intersection of recognition sequence and the binding site occupied by a single ligand molecule is not sufficient for the inhibition to occur. Restriction endonuclease cutting sites protected by netropsin can be predicted basing upon known nucleotide sequence specificity of netropsin. Netropsin and bis-netropsins show different nucleotide sequence specificity. This fact can be used for selective inhibition of restriction endonucleases. PMID- 3027537 TI - [The effect of the phage lambda ral gene on the level of synthesis of the EcoK restriction endonuclease beta-subunit]. AB - E. coli hsd genes were subcloned from lambda 642 (ral+) into lambda L47.1 vector (ral-after replacement). The influence of bacteriophage lambda ral gene on the expression efficiency of hsdS kappa, hsdM kappa genes was investigated. It was shown, that its presence in vitro enhanced the synthesis of beta-subunit, hsdM gene product, and the increase of modification in vivo was observed. It is proposed that the increase of modification rate of lambda phage fully unmodified DNA is connected with the appearance of E. coli DNA methylase consisting of beta- and gamma-subunits but lacking alpha-subunit. PMID- 3027538 TI - [Xanthogranulomatous pyelonephritis]. AB - An 11-year-old boy suffered from malaise, weight loss and pallor. A palpable abdominal tumor on the right side, anemia and increased C-reactive protein were detected. Intravenous urography revealed destruction of the right kidney resembling Wilms tumor. But ultrasound and computered tomography rised skepticism. Analysis of previously documented cases suggests that xanthogranulomatous pyelonephritis must equally be considered in a child with unilaterally enlarged kidney without function, especially when the child shows fever, leukocytosis, bacteriuria, anemia, leukocyturia, calculi of the urinary tract, abdominal pain and/or a palpable abdominal tumor. Ultrasound and computered tomography can lead to the diagnosis, and identify extrarenal infiltration. Nephrectomy results in complete cure and is therefore the treatment of choice. PMID- 3027539 TI - The microcellular (oat cell, small cell anaplastic) carcinoma of the lung. A histopathological study of 620 diagnosed cases. PMID- 3027540 TI - The etiologic agent of AIDS. PMID- 3027541 TI - Neurologic effects of HTLV-III infection in adults: an overview. PMID- 3027543 TI - Malignant lymphomas, AIDS, and the pathogenic role of Epstein-Barr virus. PMID- 3027544 TI - Toward a vaccine against AIDS: rationale and current progress. PMID- 3027542 TI - Is cytomegalovirus a cofactor in the pathogenesis of AIDS and Kaposi's sarcoma? PMID- 3027545 TI - Antiviral chemotherapies directed against HTLV-III/LAV. PMID- 3027546 TI - Diagnostic and prognostic factors in AIDS. PMID- 3027547 TI - Cytomegalovirus endometritis in a patient with AIDS. PMID- 3027548 TI - Cytomegalovirus-induced adrenal insufficiency and associated renal cell carcinoma in AIDS. PMID- 3027549 TI - [Effect of the function groups of a silane coupling agent on adhesion]. PMID- 3027550 TI - [Experimental study on the fracture toughness of composite resins]. PMID- 3027551 TI - [Molecular-genetic organization of plasmid R89S of the incompatibility group Q]. AB - A new IncQ plasmid R89S has been analysed by molecular-genetic methods. A restriction map of this plasmid has been constructed and regions of homology with the plasmid RSF1010 have been identified. A genetic map of the plasmid R89S has been prepared based on the deletion and insertion plasmid derivatives. The phenotypic analysis of the derivatives has identified the location of genes coding for replication, incompatibility, mobilization for genetic transfer and resistance to streptomycin in the genome of R89S. PMID- 3027552 TI - [Heterogeneity of nuclear proteins from HeLa cells specifically binding to the Alu sequence]. AB - Nuclear proteins from HeLa cells specifically binding to the Alu-repeat cloned in the plasmid Blur8 have been studied. 0.35 M nuclear extract proteins have been separated on DEAE-cellulose. The presence of DNA-binding proteins has been found in all fractions by the technique of DNA-binding on nitrocellulose filters. The labelled restricted DNA of the plasmid Blur8 was incubated with the proteins of different fractions with the subsequent identification of specific Alu-protein complexes in polyacrylamide gel at low ionic strength. At least two proteins have been found to have the different affinity to Alu-repeat. Various functions of Alu repeats and the possibility of their participation in the initiation of DNA replication are discussed. PMID- 3027553 TI - [Cloning of mRNA nucleotide sequences amplified during DNA replication in the regenerating liver]. AB - A library of double-stranded cDNA has been constructed using the mRNA of regenerating rat liver 20 hr after partial hepatectomy. The differential screening of the library with the regenerating liver specific and the resting liver-specific single-stranded cDNA probes has identified 11 cDNA clones which sequences are preferentially expressed in regenerating rat liver. The RNA dot blot hybridization has shown that levels of RNA complementary to these clones are 3 to 8-fold higher in dividing cells as compared with resting cells. PMID- 3027554 TI - [Structural-functional organization of R-plasmid pBS222 with a broad range of bacterial hosts]. AB - The broad host-range plasmid pBS222 is compatible with broad host-range plasmids of all known incompatibility groups and codes for tetracycline resistance. pBS222 is efficiently mobilized by Inc P-1 plasmid RP4 and is also capable of conjugal transfer with low efficiency to different gramnegative microorganisms. The size of the plasmid (17.2 Kb) has been determined and its physical map has been constructed. The plasmid harbours the unique sites for restriction endonucleases BglII, HindIII, HpaI, KpnI, SmaI and XbaI cleawage. The plasmid derivatives pBS352-pBS355 have been obtained that carry kan- and cam-determinants in addition to tet-gene. Plasmid pBS355 has been used to clone EcoRI-fragments of phage lambda DNA. The plasmid pBS222 regions essential for replication and maintenance have been localized by DNA hybridization analysis of its mini-derivatives pBS356 and 357. pBS222 is a convenient model for investigations of the plasmid replication and maintenance mechanisms in different bacterial hosts as well as for the construction of broad host-range vectors. PMID- 3027555 TI - Strain-specific 1.7 kilobase repetitive deoxyribonucleic acid sequence family in Trichinella spiralis. AB - Eco RI digestion of bulk DNA from Trichinella spiralis P1, an isolate from domestic pig, reveals the presence of families of repetitive sequences. One of the most prominent of these has a monomer size of 1.7 kb, which exists in minimally dispersed direct tandem arrays, with a copy number of about 2800, and represents 2% of the genome. Although there is evidence that the Eco RI site is missing in some of the family members and that a 1.9 kb variant of the sequence also occurs, the family is highly homogeneous. When bulk DNA from other pig isolates (P2, PB1) and two black bear isolates from Pennsylvania (UPB6, UPB8) is probed with a typical member of the 1.7 kb sequence family cloned into pUC9, hybridization is identical in pattern and intensity with self-hybridization, indicating that the 1.7 kb family exists equally in the repetitive fraction of all of these isolates. When blots of bulk DNA from wild carnivore isolates (MSIL, PF1, AF1, AF2, AF3, AF4, SL, TC) are probed with pPRA, no hybridization can be detected. Faint hybridization of the probe to DNA from T. spiralis var. pseudospiralis occurs at 1.7 kb on an Eco RI profile and indicates that the 1.7 kb sequence has been conserved in the course of strain evolution. PMID- 3027556 TI - Possible role of cAMP in the differentiation of Trypanosoma cruzi. AB - To assess the possible action of cAMP on the cell differentiation of Trypanosoma cruzi, we determined both cAMP levels and cAMP-binding activities of epimastigotes and trypomastigotes of this parasite. Trypomastigotes showed a 4 fold higher cAMP content and a 2.5-fold increase in the specific activity of a cAMP-binding protein with identical properties to that of epimastigotes. The high levels of cAMP present in trypomastigotes strongly suggest a role of this cyclic nucleotide on the differentiation of T. cruzi. PMID- 3027557 TI - Evidence of an endogenous digitalis-like factor in the plasma of patients with acromegaly. AB - Evidence suggests that plasma-volume expansion leads to the release of a digitalis-like factor, which is thought to act on the renal tubular cells and cause natriuresis. We postulated that this factor might be present in patients with acromegaly (in whom plasma volume is elevated) and might return to normal levels when the disease was treated successfully. We measured the ability of plasma extracts from patients with acromegaly to inhibit the binding of ouabain to the sodium pump in normal red cells and to inhibit the enzymatic activity (sodium-potassium-ATPase) of the sodium pump in membrane preparations from normal kidneys. In 21 patients with active acromegaly, the mean (+/- SE) level of ouabain-binding inhibition (1.56 +/- 0.38) was higher (P less than 0.01) than that in either 11 successfully treated patients (0.18 +/- 0.05) or in 27 normal controls (0.19 +/- 0.03). The inhibition of sodium-potassium-ATPase activity by plasma was also greater in patients with active acromegaly (38.1 +/- 6.8 percent) than in successfully treated patients (18.4 +/- 5.6 percent, P less than 0.05) or controls (21.1 +/- 2.7 percent, P less than 0.05). Significant correlations were found between plasma volume and ouabain-binding inhibition in 23 patients (r = 0.72, P less than 0.01) and sodium-potassium-ATPase inhibition in 19 patients (r = 0.62, P less than 0.01). Pituitary adenomectomy decreased plasma volume and the inhibition by plasma of ouabain binding. We conclude that an endogenous digitalis like factor is present in the plasma of patients with chronic volume expansion due to acromegaly. These results are consistent with the hypothesis that this natriuretic factor may have a physiologic role in water and sodium homeostasis. PMID- 3027559 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 10-1987. A 59-year-old woman with progressive polyneuropathy and monoclonal gammopathy. PMID- 3027558 TI - Sorbitol, phosphoinositides, and sodium-potassium-ATPase in the pathogenesis of diabetic complications. AB - During the past decade, our appreciation of the original experiments with myo inositol supplementation in diabetic rats has greatly expanded. The effects of myo-inositol on nerve conduction are now explained by concepts that were largely unappreciated in 1976, including the fundamental role of phosphoinositide metabolism in cell regulation and the importance of the activity of sodium potassium-ATPase in nerve conduction. Aldose reductase inhibitors firmly link defects in myo-inositol metabolism to activation of the polyol pathway in diabetes; the resulting "sorbitol-myo-inositol hypothesis" has been extended from its application to the lenses and peripheral nerves to most of the tissues involved with diabetic complications. These biochemical mechanisms provide a new framework within which to explore the complex interactions between hyperglycemia and the vascular, genetic, and environmental variables in the pathogenesis of diabetic complications. It is anticipated that these endeavors will result in the appearance of new classes of therapeutic agents, the first of which--the aldose reductase inhibitors--has emerged from the laboratory and is now undergoing extensive clinical testing. These efforts are very likely to result in the appearance of new treatment methods that may dramatically lighten the burden of chronic complications in patients with diabetes. PMID- 3027560 TI - Endogenous digitalis-like factor or factors. PMID- 3027561 TI - More on fish oil. PMID- 3027562 TI - Cerebral uptake of benzodiazepine measured by positron emission tomography in hepatic encephalopathy. PMID- 3027563 TI - Surfer's neuropathy. PMID- 3027564 TI - Signal-transduction. PMID- 3027565 TI - Tumour necrosis factor and lymphotoxin genes map close to H-2D in the mouse major histocompatibility complex. AB - Tumour necrosis factor (TNF-alpha) and lymphotoxin (TNF-beta) are related proteins, secreted by macrophages and lymphocytes respectively, which play a role in destruction of tumour cells and virally infected cells (for reviews see refs 1,2). TNF-alpha is a non-glycosylated protein of relative molecular mass 17,000 (Mr 17 K), whereas TNF-beta is a glycoprotein of Mr 25 K. Both TNF-alpha and TNF beta aggregate into multimers and act through the same receptor molecule on target cells. Genes encoding these two TNF proteins have been cloned from mouse and man and in both are closely linked, being separated by approximately 1 kilobase (kb) of DNA. In the mouse these genes are located on chromosome 17, but in man they are on the short arm of chromosome 6. This segment of chromosome 6 also contains the genes of the major histocompatibility complex (MHC), as does chromosome 17 in the mouse. To find out whether the TNF genes are located within the MHC, we used polymorphic restriction sites to analyse a panel of MHC congeneic and intra-MHC recombinant mouse strains. Initially, we mapped the TNF genes the D or Qa region in the distal half of the mouse MHC. We then studied a gene cluster encompassing part of the D and Qa regions and found the TNF genes are located 70 kb proximal to the D gene. PMID- 3027566 TI - A transcription factor which binds to the enhancers of SV40, immunoglobulin heavy chain and U2 snRNA genes. AB - In eukaryotes the transcriptional control of RNA polymerase II-mediated gene expression is exerted by cis-acting regulatory DNA elements classified as promoter and enhancer sequences. These elements are composed of a number of different protein binding sites. The regulatory factors that recognize such 'modules' may be ubiquitous, tissue- or stage-specific, and positively or negatively acting. According to this model the transcriptional activity of a given gene is programmed by a combination of different modules. We analysed such a site of protein-DNA interaction, the octamer motif, in the enhancers of the simian virus (SV40) early genes and the murine immunoglobulin heavy-chain gene, and in the distal sequence element (DSE) of the U2 small nuclear (sn)RNA gene of Xenopus laevis. The corresponding DNA-binding factor appears to be the same in the three cases. Moreover, a fraction containing partially purified octamer motif binding factor has a stimulatory effect on transcription in an in vitro system. PMID- 3027567 TI - Construction and use of human chromosome jumping libraries from NotI-digested DNA. AB - A basic difficulty in the molecular analysis of genes identified by mutations in the mammalian genome is the need to cover genetic distances corresponding to several hundred kilobases or more by molecular techniques like chromosome walking. In chromosome jumping, this limitation is overcome by the deletion of all but the extreme ends of large DNA molecules before cloning. We describe here the construction and characterization of a NotI 'jumping library' from human DNA. To characterize this library, random clones were analysed by restriction mapping. Clones carrying unique end fragments were characterized further by hybridization to Southern blots of NotI-cleaved human DNA separated on pulsed field gradient (PFG) gels. As a first step in a directional walk, the library was screened with a clone containing a NotI site cleaved in genomic DNA ('NotI linking clone') localized to the distal third of the short arm of human chromosome 4 (A.-M.F. & T.P., unpublished data). Starting and end points of two identified clones were positioned within a restriction map covering 850 kilobases. PMID- 3027568 TI - Elevated levels of diacylglycerol and decreased phorbol ester sensitivity in ras transformed fibroblasts. AB - Diacylglycerol (DG) plays a central role in phospholipid metabolism and is an endogenous activator of protein kinase C. We have suggested that constitutive activation of this kinase is one mechanism by which oncogenes transform cells. The ras-encoded proteins are similar to regulatory G-proteins and are candidates for the unknown G-protein that modulates phosphatidylinositol (PI) turnover. Differences in polyphosphoinositide metabolism have been reported for ras transformed cells. But because these experiments were performed on confluent cultures of established cell lines, the differences are difficult to attribute to ras transformation. Here we show that exponentially growing NIH 3T3 fibroblasts recently transformed by Ha-ras or Ki-ras possess elevated DG concentrations without significant alterations in the levels of other polyphosphoinositide metabolites. The basal phosphorylation of protein kinase C substrate of relative molecular mass (Mr) 80,000 (80K) is significantly increased in all the ras transformed cell lines. Surprisingly, however, further phosphorylation of this protein on addition of phorbol ester was greatly reduced. Ha-ras cells also show less binding of phorbol ester than control cells, suggesting that elevation of DG causes partial down-regulation in addition to activation of protein kinase C. PMID- 3027569 TI - Modulation of gelsolin function by phosphatidylinositol 4,5-bisphosphate. AB - The actin-binding protein gelsolin requires micromolar concentrations of calcium ions to sever actin filaments, to potentiate its binding to the end of the filament and to promote the polymerization of monomeric actin into filaments. Because transient increases in both intracellular [Ca2+] and actin polymerization accompany the cellular response to certain stimuli, it has been suggested that gelsolin regulates the reversible assembly of actin filaments that accompanies such cellular activations. But other evidence suggests that these activities do not need increased cytoplasmic [Ca2+] and that once actin-gelsolin complexes form in the presence of Ca2+ in vitro, removal of free Ca2+ causes dissociation of only one of two bound actin monomers from gelsolin and the resultant binary complexes cannot sever actin filaments. The finding that cellular gelsolin-actin complexes can be dissociated suggests that a Ca2+-independent regulation of gelsolin also occurs. Here we show that, like the dissociation of profilin-actin complexes, phosphatidylinositol 4,5-bisphosphate, which undergoes rapid turnover during cell stimulation, strongly inhibits the actin filament-severing properties of gelsolin, inhibits less strongly the nucleating ability of this protein and restores the potential for filament-severing activity to gelsolin-actin complexes. PMID- 3027570 TI - Activation of transcription by two factors that bind promoter and enhancer sequences of the human metallothionein gene and SV40. AB - Genetic analysis of eukaryotic transcriptional promoters has revealed that protein-coding genes often contain a complex array of cis-control elements consisting of upstream activator sequences and enhancer elements. The metallothionein genes provide a useful example for dissecting the action of multiple interspersed control elements that govern both basal level and regulated expression in animal cells. The human metallothionein (hMTIIA) promoter has been analysed in detail and found to contain no less than five distinct control elements in the 5' flanking regions of the gene that mediate specificity and regulation of transcription. These different control elements can be functionally subdivided into two categories: basal and induced elements. There are several distinct basal recognition sequences, which include a TATA-box, a GC-box, and at least two basal level enhancer (BLE) sequences, that function like classical enhancer elements. The hMTIIA gene also responds to induction by heavy metals and by steroid hormones through the action of metal regulatory elements (MRE) and glucocorticoid responsive elements (GRE). Here we report the identification of two cellular DNA-binding proteins that interact selectively with sequences governing the basal level expression of hMTIIA. One of these factors is a novel activator protein (AP1) that interacts with sequences in the BLE of hMTIIA and also binds to a site within the 72-base pair (bp) repeats of the simian virus 40 (SV40) enhancer region. The second protein has been purified to homogeneity and shown to be transcription factor Sp1 which recognizes and binds to a single GC box element within the hMTIIA promoter. PMID- 3027571 TI - Another cyclic-nucleotide-gated conductance. PMID- 3027572 TI - Communication between segments of DNA during site-specific recombination. AB - Some site-specific recombination systems require the interacting DNA sequences to have a specific relative orientation. This means that DNA segments can sense each other's direction even though they may be separated by many thousands of base pairs. Here, we review the surprising results of recent experiments that lead to a new model which accounts for site orientation specificity and relates it to other recombination systems where relative orientation is not critical. PMID- 3027573 TI - Deregulated c-fos expression interferes with normal bone development in transgenic mice. AB - Transgenic mice were generated expressing c-fos genes under the control of the human metallothionein promoter. Although high levels of c-fos messenger RNA were detectable in a variety of tissues, deregulated c-fos expression specifically interfered with normal bone development without inducing malignant tumours. PMID- 3027574 TI - A cyclic nucleotide-gated conductance in olfactory receptor cilia. AB - Olfactory transduction is thought to be initiated by the binding of odorants to specific receptor proteins in the cilia of olfactory receptor cells. The mechanism by which odorant binding could initiate membrane depolarization is unknown, but the recent discovery of an odorant-stimulated adenylate cyclase in purified olfactory cilia suggests that cyclic AMP may serve as an intracellular messenger for olfactory transduction. If so, then there might be a conductance in the ciliary plasma membrane which is controlled by cAMP. Here we report that excised patches of ciliary plasma membrane, obtained from dissociated receptor cells, contain a conductance which is gated directly by cAMP. This conductance resembles the cyclic GMP-gated conductance that mediates phototransduction in rod and cone outer segments, but differs in that it is activated by both cAMP and cGMP. Our data provide a mechanistic basis by which an odorant-stimulated increase in cyclic nucleotide concentration could lead to an increase in membrane conductance and therefore, to membrane depolarization. These data suggest a remarkable similarity between the mechanisms of olfactory and visual transduction and indicate considerable conservation of sensory transduction mechanisms. PMID- 3027575 TI - Unusual intron in the immunoglobulin domain of the newly isolated murine CD4 (L3T4) gene. AB - The T-cell surface glycoprotein, CD4, is expressed predominantly on helper T cells and is thought to play a major role in cell-cell interactions. Monoclonal antibodies against CD4 have been shown to block numerous T-cell functions; moreover, recent results suggest that the CD4 molecule may be involved in transmembrane signal transduction. The human CD4 glycoprotein has also been shown to form at least part of the receptor for the AIDS virus, HIV-1. Elucidation of the functions of CD4 will be facilitated by the ability to manipulate the protein by genetic means. Because the mouse system is well suited for a variety of functional studies, we have isolated, sequenced and expressed cDNA clones encoding the murine CD4 (L3T4) glycoprotein. Comparison of the mouse and human CD4 sequences reveals striking evolutionary conservation of the cytoplasmic domain, suggesting that this region is essential for CD4 function. In addition, both the human and mouse CD4 gene contain a large intron in the coding region of the V-like domain. As no other members of the immunoglobulin gene superfamily have been shown to contain similarly placed introns, this finding may have important implications regarding the evolution of this gene family in particular and of introns in general. PMID- 3027576 TI - Activation of sodium-proton exchange is a prerequisite for Ca2+ mobilization in human platelets. AB - Stimulated platelets take up sodium ions and release hydrogen ions due to activation of Na+/H+ exchange resulting in cytoplasmic alkalinization. Suppression of Na+/H+ exchange either by removal of extracellular Na+ or by application of amiloride inhibits shape change, secretion of granule contents and aggregation. The data we present here indicate that inhibition of this transport by ethylisopropyl-amiloride or by lowering extracellular sodium reduces or even completely suppresses the rise in cytoplasmic free Ca2+ concentration that is essential for platelet aggregation in response to thrombin. We also demonstrate that cytoplasmic alkalinization produced by exposure to the ionophore monensin sensitizes the human platelet response to stimulation by thrombin resulting in enhanced Ca2+ mobilization and aggregability. We conclude that an increase in intracellular pH evoked by activation of Na+/H+ counter transport is an important signal in stimulus-response coupling and forms an essential step in the cascade of events required to increase cytoplasmic free Ca2+ in platelets. PMID- 3027578 TI - Herpes infection and AIDS. PMID- 3027577 TI - Gene structure and extracellular secretion of Neisseria gonorrhoeae IgA protease. AB - Several human bacterial pathogens, including the Gram-negative diplococcus Neisseria gonorrhoeae, produce extracellular proteases that are specific for human immunoglobulin IgA1. Immunoglobulin A (IgA) proteases have been studied extensively and the genes of some species cloned in Escherichia coli, but their role in pathogenesis remains unclear. Recently we derived a DNA fragment of 5 kilobases (kb) from N. gonorrhoeae MS11 directing extracellular active enzyme in E. coli. Although the mature enzyme of strain MS11 was shown to have a relative molecular mass of 106,000 (Mr 106K) in gels, the DNA sequence of this cloned fragment reveals a single gene coding for a 169K precursor of IgA protease. The precursor contains three functional domains, the amino-terminal leader which is assumed to initiate the inner membrane transport of the precursor, the protease, and a carboxyl-terminal 'helper' domain apparently required for extracellular secretion (excretion). Based on the structural features of the precursor, we propose a model in which the helper serves as a pore for excretion of the protease domain through the outer membrane. IgA protease acquires an active conformation as its extracellular transport proceeds and is released as a proform from the membrane-bound helper by autoproteolysis. The soluble proform further matures into the 106 K IgA protease and a small stable alpha-protein. PMID- 3027579 TI - Transforming potential of the c-fms proto-oncogene (CSF-1 receptor). AB - The c-fms proto-oncogene encodes a transmembrane glycoprotein that is probably identical to the receptor for the macrophage colony stimulating factor, CSF-1. Forty C-terminal amino acids of the normal receptor are replaced by 11 unrelated residues in the feline v-fms oncogene product, deleting a C-terminal tyrosine residue (Tyr969) whose phosphorylation might negatively regulate the receptor kinase activity. We show that the human c-fms gene stimulates growth of mouse NIH 3T3 cells in agar in response to human recombinant CSF-1, indicating that receptor transduction is sufficient to induce a CSF-1 responsive phenotype. Although cells transfected with c-fms genes containing either Tyr969 or Phe969 were not transformed, cotransfection of these genes with CSF-1 complementary DNA induced transformation, with c-fms(Phe969) showing significantly more activity than c-fms(Tyr969). In the absence of CSF-1, chimaeric v-fms/c-fms genes encoding the wild-type c-fms C terminus were poorly transforming, whereas chimaeras bearing Phe969 were as transforming as v-fms. Thus, the Phe969 mutation, although not in itself sufficient to induce transformation, activates the oncogenic potential of c-fms in association with an endogenous ligand or in conjunction with mutations elsewhere in the c-fms gene that confer ligand-independent signals for growth. PMID- 3027580 TI - Gene transfer and molecular cloning of the rat nerve growth factor receptor. PMID- 3027581 TI - A novel abl protein expressed in Philadelphia chromosome positive acute lymphoblastic leukaemia. AB - The Philadelphia (Ph) chromosome breakpoints in chronic myelocytic leukaemia are clustered on chromosome 22 band q11 in a 5.8-kilobase (kb) region designated bcr. The c-abl protooncogene is translocated from chromosome 9 band q34 into bcr and the biochemical consequence of this molecular rearrangement is the production of an abnormal fusion protein bcr-abl p210 with enhanced protein-tyrosine kinase activity compared to the normal p145 c-abl protein. The Ph chromosome translocation is also seen in some acute lymphoblastic leukaemias with B-cell precursor phenotypes some of which have bcr rearrangement (bcr+) and some do not (bcr-). We present evidence that the Ph+, bcr- leukaemias are associated with a novel p190 abl kinase. We propose that acute lymphoblastic leukaemias that are bcr+, p210+ are probably lymphoid blast crises following a clinically silent chronic phase of chronic myelocytic leukaemia arising in multipotential stem cells whereas bcr-, p190+ cases are de novo acute lymphoblastic leukaemias arising in more restricted precursors. PMID- 3027582 TI - Specific antigen-Ia activation of transfected human T cells expressing murine Ti alpha beta-human T3 receptor complexes. AB - The genes encoding the alpha and beta chains of the T-cell receptor (Ti) of an antigen-specific, Ia-restricted murine T-cell hybridoma were introduced into T3 positive or T3-negative human T cells. The resultant transfectants express either mouse-human or mouse-mouse Ti alpha beta molecules functionally associated with the human T3 complex. Only the complete murine Ti alpha beta dimer mediates specific functional corecognition of the appropriate antigen-Ia pair. PMID- 3027583 TI - Inositol trisphosphate receptor localization in brain: variable stoichiometry with protein kinase C. AB - Many neurotransmitters, hormones and growth factors act at membrane receptors to stimulate the phosphodiesteratic hydrolysis of phosphatidyl-inositol 4,5 bisphosphate generating the comessengers inositol 1,4,5-trisphosphate (Ins(1,4,5)P3) and diacylglycerol. Diacylglycerol stimulates protein kinase C3 while Ins(1,4,5)P3 is postulated to activate specific receptors leading to release of intracellular calcium, probably from the endoplasmic reticulum. In recent preliminary reports, Rubin and associates detected 32P-Ins(1,4,5)P3 binding to liver and adrenal microsomes and to permeabilized neutrophils and liver cells. We now report the biochemical and autoradiographic demonstration in brain of high affinity, selective binding sites for 3H- and 32P-labelled Ins(1,4,5)P3 at levels 100-300 times higher than those observed in peripheral tissues. The potencies of various myoinositol analogues at the Ins(1,4,5)P3 binding site correspond to their potencies in releasing calcium from microsomes, supporting the physiological relevance of this receptor. Brain autoradiograms demonstrate discrete, heterogeneous localization of Ins(1,4,5)P3 receptors. In some regions localizations of Ins(1,4,5)P3 receptors resemble those of protein kinase C14, while in others they differ markedly, suggesting a novel mechanism whereby the relative activity of the two limbs of the PI cycle can be differently regulated. PMID- 3027585 TI - Choline+: a substrate of the neuronal noradrenaline carrier in the rat vas deferens. AB - The effects of choline+ (10-40 mmol/l) on 3H-noradrenaline uptake by, and 3H noradrenaline efflux from, noradrenergic neurones were studied in vasa deferentia of reserpine-pretreated rats at an external Na+ concentration of 100 mmol/l. Monoamine oxidase and catechol-O-methyltransferase were inhibited. Choline+ (20 and 40 mmol/l) competitively inhibited the neuronal uptake of 3H-noradrenaline. From the choline+-induced changes in the apparent Km for 3H-noradrenaline transport, a Ki of 35 mmol/l was obtained. Choline+ (10, 20 and 40 mmol/l) accelerated the neuronal efflux of 3H-noradrenaline in a concentration-dependent manner. This acceleration of efflux was greatly reduced in the presence of 1 mumol/l desipramine, indicating that choline+ is capable of eliciting "accelerative exchange diffusion". Choline+ (40 mmol/l) and (-)noradrenaline (4.5 mumol/l) (i.e., concentrations about equivalent to the Ki and Km for choline+ and (-)noradrenaline, respectively) produced virtually identical increases in the neuronal efflux of 3H-noradrenaline. Choline+ (3-300 mmol/l) inhibited the specific binding of 3H-desipramine to plasma membranes derived from cultured rat phaeochromocytoma (PC-12) cells. The Ki for this interaction was 48 mmol/l. This results suggest that choline+ acts as alternative substrate of the neuronal noradrenaline transport system and should, therefore, not be used in transport studies with noradrenaline as substitute for Na+ in Na+-deficient media. PMID- 3027584 TI - Association of the polyomavirus middle-T antigen with c-yes protein. AB - Expression of the middle-T antigen of polyomavirus is sufficient to induce transformation of fibroblasts in culture and tumour formation in whole animals. Middle-T can form a complex with the cellular src gene product (p60c-src) and can be phosphorylated by p60c-src in vitro. Studies using middle-T mutants have suggested that the association of middle-T with p60c-src may be necessary but not sufficient for transformation. Therefore, we addressed the possibility that middle-T could interact with other tyrosine protein kinases structurally related to p60c-src. Using antibody raised against a fusion protein between beta galactosidase and amino-terminal sequences of p90gag-yes from Y73 virus (anti-yes antibody), we have found that middle-T can associate with and be phosphorylated by the c-yes proto-oncogene product, a protein of relative molecular mass (Mr) 62,000 (62K). This raises the possibility that the middle-T-p62c-yes complex contributes to transformation by polyomavirus. PMID- 3027588 TI - Getting up groggy. PMID- 3027586 TI - Regulation of alpha and beta adrenergic receptors by triiodothyronine in cultured rat myocardial cells. AB - Previously, in this laboratory, we established the presence of alpha and beta adrenergic receptors in primary cultures of neonatal rat heart cells. We now report that exposure to 1-triiodothyronine (10 nM) regulates the binding characteristics of these receptors. As measured by [125I]-I-2-[beta-(4 hydroxyphenyl)ethylaminomethyl]tetralone ([125I]IBE 2254), alpha receptor number (Bmax) decreased by 50% from 37,000 +/- 4,500 to 18,000 +/- 4,300 sites per cell and the equilibrium dissociation constant (KD) decreased from 420 +/- 24 to 150 +/- 37 pM. As measured by [125I]-iodocyanopindolol ([125I]ICYP), beta receptor number increased by 42% from 12,000 +/- 2,600 to 17,000 +/- 4,000 sites per cell with no accompanying change in affinity. An increase in maximal stimulation of adenylate cyclase activity by isoproterenol was also detected under conditions of excess triiodothyronine. No significant changes in agonist or antagonist affinities for either the alpha or the beta adrenergic receptor were detected in thyroid hormone treated cultures. It can be concluded that in cultured myocardial cells thyroid hormone regulates the characteristics of both alpha and beta adrenergic receptors, but in a strikingly different manner. PMID- 3027589 TI - [Microangiopathic hemolytic anemia and metastasized carcinoma]. PMID- 3027587 TI - Correlation between the negative inotropic potency and binding parameters of 1,4 dihydropyridine and phenylalkylamine calcium channel blockers in cat heart. AB - Partially purified plasma membranes were prepared from cat ventricle. The purification factors for the calcium channel ligands (+)-3H-PN 200-110, 3H nimodipine (1,4-dihydropyridines) and (-)-3-H-desmethoxyverapamil (a phenylalkylamine) were 3.1-, 3.4- and 2.9-fold, respectively, whilst beta adrenoceptors labelled with (-)-3H-dihydroalprenolol were purified 3.0-fold. (+) 3H-PN 200-110 bound to 930 +/- 140 fmol/mg of membrane protein with a dissociation constant of 70 pmol/l at 25 degrees C. Under the same conditions 3H nimodipine bound to 490 +/- 24 fmol/mg of sites with a KD of 120 pmol/l. (-)-3-H desmethoxyverapamil bound to 530 +/- 55 fmol/mg of sites with a KD of 2.47 nmol/l. Twelve 1,4-dihydropyridines were evaluated for binding inhibition constants (Ki) with (+)-3H-PN 200-110 and 13 phenylalkylamines with (-)-3-H desmethoxyverapamil in radioligand binding assays. Of the twelve 1,4 dihydropyridines evaluated (+/-)-nitrendipine was the most potent with a Ki-value of 280 pmol/l, nifedipine had a Ki-value of 500 pmol/l and the weakest drug tested, (+/-)-Bay b 4328, had a Ki-value of 14.3 nmol/l. The EC50-values of the same 1,4-dihydropyridines to inhibit the electrically driven cat papillary muscle were 77- to 3,450-fold higher and little correlation existed between Ki and EC50 values. Thirteen phenylalkylamines were tested for their potency to inhibit (-)-3 H-desmethoxyverapamil binding. The most potent phenylalkylamine with respect to negative inotropy was (+/-)-D 595 with an EC50-value of 794 nmol/l, the least potent substance was (+/-)-Sz 45 with an EC50-value of 39.8 mumol/l. The binding inhibition constants for the phenylalkylamines were 13- to 322-fold lower than the values for negative inotropy, but a significant positive correlation between the Ki and EC50-values (n = 12, r = 0.84) was observed. The absolute differences may reflect the state-dependent binding of phenylalkylamines to the channel. QSAR analysis revealed nearly identical correlations between physicochemical substituent properties on the one hand and binding affinities or functional potency on the other hand. In both cases the electronic properties (F-constant) of ring substituents mainly determine the variance in potency. PMID- 3027590 TI - [Prenatal diagnosis of cystic fibrosis using DNA analysis]. PMID- 3027591 TI - [Polymodality of the distribution of synaptic delays in the neuromuscular junctions of the frog]. AB - Measurements of the synaptic delay of uniquantal responses have been made on frog cutaneous pectoris muscle. Extracellular focal recording and low mean quantal content (0.05-1) was used. The distribution of delays obtained had a polymodal character with the mean modal interval equal to 0.22 +/- 0.01 ms (n = 13). An increase of the quantal content induced only a redistribution of the mode weights, but the modal interval did not change. A decrease in the temperature induced an increase in the modal interval with the temperature coefficient Q10 = 2.42 +/- 0.14 (n = 15). According to the hypothesis which was used for explaining the data obtained, the process of transmitter quanta release is determined by interaction of two molecular mechanisms: an increase of the release probability depending on the intracellular calcium concentration; rhythmical work of the mechanism realizing the quantum release. The latter does not depend on the calcium concentration but depends on temperature. The polymodal distribution of synaptic delays reflects this rhythmical work of each presynaptic active zone. PMID- 3027592 TI - [Effect of the locomotor system of the pedal ganglia of the pteropodial mollusk on anatomically isolated neurons]. AB - The isolated pedal ganglia of the pteropodial mollusc Clione limacina generate the locomotor activity. In 30% of the pedal ganglion preparations, the locomotor rhythm was not regular, i. e. the locomotor generator worked in "bursts". These "locomotor bursts" were caused by spontaneous activations of command neurons located in the pedal ganglia. Single neurons were extracted from such preparations with an intracellular microelectrode and then their somas were put into the initial place between the ganglion cells. 25% of the isolated neurons (9 out of 35) renewed the "locomotor bursts"-related changes in the activity after the insertion into the ganglion. Neurons, originally excited during "bursts", continued to be excited after isolation, while inhibited neurons continued to be inhibited. It follows, therefore, that the command neurons can act on the target cells in the absence of morphological synapses. PMID- 3027593 TI - [Spinal control of afferent temperature information in the cat]. AB - The picrotoxin effect on background and evoked discharges of interneurons in the superficial laminae of cat dorsal horn was investigated under mechanical and temperature influences on skin receptors. It was shown that information about skin temperature is modulated on the presynaptic level. It is concluded that competitive interaction between fibres of large and small diameters is of certain importance in the transmission of such information. PMID- 3027594 TI - [Spectrum of high-frequency oscillations of the electrocorticogram and their nature]. AB - A gradual amplitude decrease towards higher frequencies was found in the spectrum (25-400 Hz) of the ECoG of the curarized rabbit. This gradual decrease of the amplitude continued to exist under the action of d-amphetamine, physostigmine, atropine, chlorpromazine and thiopental despite changes in the spectrum characteristic of every agent. These changes allowed finding the boundary near 40 Hz between high frequencies (HF) and conventional ECoG. The amplitudes of all HF components changed in the same direction independently of the ECoG. Unanimous changes suggest common genesis of HFs and a gradual decrease of the amplitude in the spectrum supports their synaptic but not spike origin. In the light of the concept of the EEG quantum the conventional ECoG represents synchronization and HFs express fluctuations of the quantum flow. PMID- 3027595 TI - Bovine leukemia provirus in the DNA of different infected host cells. AB - Bovine leukemia provirus is reported to be integrated in the DNA of different infected mammalian cells. We observed morphological transformation in BLV infected sheep fetal spleen, kidney, thymus and sternal cultures. The presence of BLV specific sequences in their genome was established after digestion with the restriction endonuclease EcoRI and hybridization with a BLV specific probe. Human myeloma ARH77 and myeloid K562 cells infected with BLV were virus productive as detected by a reverse transcriptase assay. The presence of proviral sequences was confirmed after Southern blotting analysis. Restriction digestion by SacI enzyme yielded a complete 8.9 kb BLV provirus in infected ARH77 cells and a smaller 7.5 kb BLV fragment in infected K562 cells. PMID- 3027596 TI - [Multiple surgery of recurrent intramedullary spinal cord tumors]. PMID- 3027597 TI - Comparison of the effects of castration and steroid replacement on incertohypothalamic dopaminergic neurons in male and female rats. AB - The activities of incertohypothalamic (IH) and tuberoinfundibular (TI) dopamine (DA) neurons were compared in selected brain regions of male and female rats by measuring the rate of DA turnover (alpha-methyltyrosine-induced decline in brain DA concentrations). The rates of DA turnover in regions containing TIDA (median eminence) and rostral IHDA (rostral periventricular and medial preoptic nuclei) neurons were greater in diestrous females than in intact males. In contrast, the rate of DA turnover in the caudal IHDA neurons (medial zona incerta), was greater in intact males than diestrous females. These results indicate that the activities of IHDA neurons, like those of TIDA neurons, differ between the sexes but that the sexual differentiation of IHDA neurons is not homogeneous. Two weeks following orchidectomy, the rates of DA turnover were increased in the median eminence and decreased in the medial preoptic nucleus. Testosterone replacement in orchidectomized males produced opposite effects, causing a decrease in DA turnover in the median eminence and an increase in the medial preoptic nucleus. In female rats, the rates of DA turnover were decreased in the median eminence and medial zona incerta and increased in the medial preoptic nucleus 2 weeks following ovariectomy. Only in the median eminence did 2 days of estrogen replacement in ovariectomized rats produce effects opposite those seen after ovariectomy alone. These data show that the activities of IHDA neurons, as estimated from measurements of DA turnover, can be altered by the removal and replacement of the gonadal steroids. PMID- 3027598 TI - Copper stimulation of LHRH release from median eminence explants. III. A process dependent on extracellular sodium. AB - Copper, complexed to histidine (CuHis), stimulates LHRH release from explants of the median eminence area (MEA). To gain further understanding of the mechanism of copper action, in this study, we assessed the Na+ and energy requirements for CuHis stimulation of LHRH release. MEA explants, obtained from adult male rats, were incubated at 37 degrees C for 15 min with 100 microM CuHis and then for 45 min in CuHis-free medium (Krebs-Ringer-phosphate buffer, pH 7.4). LHRH released into the medium was evaluated by RIA. When the incubation buffer contained 143 mM Na+, CuHis stimulated the release of LHRH from a basal level of 17.2 +/- 1.26 (mean +/- SEM, n = 7) to 74.5 +/- 6.2 pg/60 min per MEA. When [Na+] was reduced to 16 mM Na+ (by substituting with Li+), CuHis-stimulated LHRH release was inhibited by 80% (p less than 0.001); indicating a requirement for Na+. In addition, we found that CuHis-stimulated LHRH release was a saturable function of Na+ concentration; saturation achieved with about 100 mM Na+. To assess the requirement for Na+ transport, we evaluated the effect of 1 mM ouabain, 10 microM tetrodotoxin (TTX), or 100 microM amiloride on CuHis stimulation of LHRH release. Ouabain inhibited CuHis stimulation of LHRH release by 80%, whereas TTX and amiloride were ineffective. In addition, we observed that CuHis did not stimulate LHRH release when incubation was carried out at 4 degrees C or at 37 degrees C in the presence of 5 mM KCN.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3027599 TI - Role of oxytocin in the modulation of ACTH release in women. AB - To determine if oxytocin (OT) may have a modulatory role on corticotropin releasing factor (CRF) and vasopressin (AVP) mediated ACTH-cortisol release in women, serial experiments were performed in which saline, OT, AVP and CRF were administered singly or in combinations. OT administration (2 IU intravenous bolus followed by 111 mIU/min infusion for 3 h) maintained a circulating concentration of 7.7 X 10(-8) M and did not significantly influence basal, AVP or CRF-induced ACTH-cortisol release. In contrast, OT inhibited significantly the potentiating effect of AVP on CRF-stimulated ACTH-cortisol release. These findings suggest that OT and AVP may modulate, in a reciprocal fashion, the CRF-mediated ACTH release and support the contention that OT may be involved in the neuroendocrine response to stress in women. PMID- 3027601 TI - Effects of sodium valproate and diazepam on beta-endorphin, beta-lipotropin and cortisol secretion induced by hypoglycemic stress in humans. AB - Evidence that gamma-aminobutyric acid (GABA) and benzodiazepine receptors play a role in the inhibition of ACTH-cortisol secretion in humans has until now been drawn only from data indicating that sodium valproate, a GABA mimetic, and diazepam, a benzodiazepine, decrease hypothalamus-pituitary-adrenal (HPA) axis secretion in patients affected by pathological hypersecretion of the axis. Therefore, the present study investigated the effects, in the same healthy subjects, of sodium valproate or diazepam, on both basal and stress-stimulated concentrations of beta-endorphin (beta-EP), beta-lipotropin (beta-LPH) and cortisol. A single maximal dose of sodium valproate (400 mg) or diazepam (10 mg) did not significantly modify basal concentrations of beta-EP, beta-LPH and cortisol. On the other hand, in the same subjects, pretreatment with sodium valproate (20 mg X 3) or diazepam (10 mg X 2) blocked the increases in these hormones produced by hypoglycemic stress in all patients tested (p less than 0.01 vs. placebo at 45, 60 and 90 min after insulin injection), without affecting the decrease in blood glucose levels. The present data show that sodium valproate and diazepam inhibit stress-induced beta-EP, beta-LPH and cortisol secretion in humans, suggesting that endogenous GABA and benzodiazepine receptors participate in physiological mechanisms regulating the activity of the HPA axis. PMID- 3027600 TI - Opioid modulation of LHRH release in vitro depends upon levels of testosterone in vivo. AB - The in vitro release of LHRH from hypothalami of adult male rats (intact, 5-day castrates, 5-day castrates replaced with various doses of testosterone) was measured under basal conditions and after the addition of KCl, the opiate antagonist naloxone or the opiate agonist DAGO to the perifusion medium. Hypothalami from all treatment groups responded to 56 mM KCl with an increased output of LHRH. LHRH release was also induced by naloxone (10(-6)M), but only from tissues derived from intacts and castrates given physiological doses of testosterone. The opiate agonist DAGO (10(-6)M) did not alter the basal release of LHRH; it, however, caused a significant decrease in the K+-induced release of LHRH from hypothalami derived from intact rats and rats replaced with physiological levels of testosterone but not from those derived from castrate rats or castrate rats replaced with small amounts of testosterone. The specificity of this latter response was shown by its reversibility with naloxone. The lack of DAGO effects upon tissues from rats with low levels of steroid implied steroid dependency of the response to opioidergic influences and indeed, the response to DAGO was restored when testosterone was replaced at physiological doses. Measurement of hypothalamic LHRH content showed no significant differences between tissues obtained from intact, castrate and testosterone-replaced castrate rats. These in vitro data support the view that the inhibitory influence of opioids upon LHRH release depends on the presence of gonadal steroids in vivo. PMID- 3027602 TI - Interaction of rat adenohypophyseal vasopressin receptors with vasopressin analogues substituted at positions 7 and 1: dissimilarity from the V1 vasopressin receptor. AB - We readdressed the question of whether or not rat adenohypophyseal vasopressin receptors have a ligand selectivity which is similar to that of the V1 subtype of vasopressin receptors. Vasopressin analogues substituted in positions 7 and 1 were used. By incubating rat anterior pituitary quarters or by perifusing rat isolated anterior pituitary cells, the effect of the vasopressin analogues on the release of beta-endorphin-like or adrenocorticotropin-like immunoreactivity was examined. The replacement of the proline residue in position 7 by sarcosine or N methyl-alanine did not change the maximum effect reached but increased the EC50 values 20- or 5-fold, respectively, when compared with arginine vasopressin. This decrease in beta-endorphin-releasing activity was no longer observed after additional removal of the alpha-amino group of cysteine in position 1. Since these substitutions are known to drastically reduce vasopressor activity, these data suggest that the beta-endorphin-releasing activity of vasopressin can be dissociated from its V1 receptor activity. Vasopressin analogues substituted in position 7 and with deaminopenicillamine or beta-mercapto-beta,beta cyclopentamethylenepropionic acid in position 1 were found to be weak antagonists of the beta-endorphin-releasing activity of vasopressin. Since these analogues are potent antagonists at the V1 receptor, these data suggest that the deaminopenicillamine and, more so, the beta-mercapto-beta,beta cyclopentamethylenepropionic acid residues in position 1 of vasopressin are strong 'binding elements' at the V1 vasopressin receptor but weak 'binding elements' at the adenohypophyseal vasopressin receptor.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3027603 TI - Participation of the central amygdaloid nucleus in the response of adrenocorticotropin secretion to immobilization stress: opposing roles of the noradrenergic and dopaminergic systems. AB - Bilateral lesions of the amygdaloid central nucleus (ACE) significantly diminished the secretion of ACTH in response to immobilization stress. Stress is associated with increased noradrenergic activity in the ACE and in the anterior and lateral hypothalamic areas. In comparison with intact stressed animals, lesion of the ACE reduced the noradrenergic activity in response to stress within the anterior and lateral hypothalamic areas, the arcuate and paraventricular nuclei of the hypothalamus and the bed nucleus of the stria terminalis. These results support the hypothesis of a stimulatory role for the noradrenergic system in the ACE on ACTH secretion. Stress decreased dopaminergic activity in the ACE, the cortical nucleus of the amygdala, the dorsomedial and ventromedial nuclei of the hypothalamus and the ventral tegmental area. In comparison with intact stressed rats, lesion of the ACE reduced dopaminergic activity in the anterior and lateral hypothalamic areas. Our results support the hypothesis of an inhibitory role of the dopaminergic system, particularly in the ACE, on ACTH secretion. This study also indicates that, in the control of ACTH secretion in response to immobilization stress, the noradrenergic and dopaminergic systems act in opposition to one another in certain brain structures such as the anterior and lateral hypothalamic areas and the ACE. PMID- 3027605 TI - Herpes simplex type II encephalitis in infancy presenting with focal encephalitis. AB - Herpes virus was recovered from a throat swab, nasopharyngeal washings as well as from brain tissue from a six-month-old infant, who presented with fever, left focal seizures and an enhancing right frontal CT-scan lesion. Cytopathic effect (CPE) as seen with genital herpetic infection was seen, suggesting HSV-2. Immunofluorescent typing of the virus isolate confirmed HSV-2. Early IgM positivity preceding a CF and SN titer rise was observed. The patient received a course of ARA-A and recovered with a left sided hemiparesis. HSV-2 encephalitis occurs beyond the newborn period as a primary infection. In adults however HSV-2 encephalitis occurs predominantly in the immunocompromised host. HSV-1 encephalitis probably is due to reactivation of a latent herpetic infection in previous virus exposed juvenile or adult immunocompetent hosts. PMID- 3027604 TI - Alpha-adrenergic stimulation of corticotropin secretion by a specific central mechanism in man. AB - In a double-blind study in normal subjects, methoxamine, a highly selective agonist at alpha-1-adrenoceptors, significantly increased circulating ACTH and cortisol. The stimulant effect of methoxamine on cortisol secretion was dose dependent in the range 3.5-7 micrograms/kg/min, was abolished by concomitant administration of the strong alpha-1-adrenergic (and weak H1) antagonist thymoxamine but unaffected by the antihistamine, chlorpheniramine. In order to test whether the action of methoxamine on ACTH secretion was exerted centrally or peripherally, the effects of norepinephrine (NE), an alpha-1-agonist that does not cross the blood-brain barrier, were studied. Doses of NE (1-12 micrograms/min) that increased systolic blood pressure by amounts similar to the changes produced by methoxamine, did not result in any rise in plasma cortisol in normal subjects. The effect of methoxamine, which is more lipid soluble than NE, on plasma ACTH and cortisol, appears to be exerted on the CNS and not at the pituitary or via nonspecific peripheral mechanisms. In addition to its water solubility, NE differs from methoxamine in its beta-1-, beta-2- and alpha-2 agonist actions. However, prenalterol (2 mg) and salbutamol (250 micrograms), respectively beta-1- and beta-2-adrenergic agonist drugs, had no effect on the secretion of ACTH or cortisol and the alpha-2-antagonist yohimbine in an effective dose did not unmask a stimulant effect of intravenous NE on plasma cortisol. At high infusion rates, NE significantly inhibited cortisol secretion. Stimulation of central alpha-1-adrenergic mechanisms results in secretion of ACTH in man, presumably by increased release of a corticotropin-releasing factor. PMID- 3027606 TI - Benign mitochondrial myopathy with deficiency of NADH-CoQ reductase and cytochrome c oxidase. AB - Since birth a female child had been weak and hypotonic. At three months of age, head control was lacking; sucking and crying were poor. Four months later, there were more spontaneous movements and the girl was able to push herself up in prone position. Further motor improvement was noted at the age of 15 months. A 25-year old brother of the patient's mother was very floppy during early childhood and has still some difficulties to swallow. Laboratory work-up showed elevated blood lactate and pyruvate levels, a mild hyperalaninemia and hyperalaninuria and an increased urinary excretion of dicarboxylic acids. Light and electron microscopy of a muscle biopsy disclosed a mitochondria-lipid-glycogen myopathy. Biochemical studies on a second muscle specimen revealed a combined deficiency of NADH-CoQ reductase and cytochrome c oxidase with a low carnitine level. There exists a considerable clinical and biochemical heterogeneity among the myopathies due to disturbances in the mitochondrial respiratory chain. PMID- 3027607 TI - Respiratory patterns in human brain tumors. AB - It has been recognized for some time that malignant cells have a diminished respiratory rate coupled with an excessive rate of aerobic glycolysis. Subsequent studies indicated, however, that this pattern was neither unique to malignant tumors nor an essential characteristic of all varieties of cancer. In attempting to synthesize the data on oxygen consumption of human brain tumors, it is difficult to relate activity to malignancy, and the integrity of the mitochondrial electron transport chain has not been systematically examined. In this study, oxygen consumption was measured using a whole cell mixture and subsequently with isolated mitochondria prepared by routine differential centrifugation from a variety of human brain tumors. It is clear from these data that low oxidative respiration is not uniquely characteristic of malignant tumors, but that a number of benign tumors such as meningiomas and pituitary adenomas display very low levels of oxygen consumption. Many of these tumors have normal respiration, with isolated mitochondria using substrates that enter at different points in the electron transport pathway. However, several of the tumors in this series showed defects in respiration at various points in the electron transport pathway. These data suggest that both benign and malignant brain tumors have depressed respiratory capacities secondary to either a decrease in mitochondria per cell or defects in electron transport activities. PMID- 3027608 TI - Pediatric central nervous system tumors: a cell kinetic study with bromodeoxyuridine. AB - Bromodeoxyuridine (BrdU), 150 to 200 mg/m2, was administered at the time of operation to 20 pediatric patients with neuroectodermal tumors to label tumor cells in the S phase. Immunocytochemical techniques were used on excised tumor specimens to detect cells containing BrdU, and the BrdU labeling index (LI) was calculated as the number of BrdU-labeled cells divided by the total number of cells counted. Four medulloblastomas, three glioblastomas multiforme, and two highly anaplastic astrocytomas had average BrdU LIs of 13.0 +/- 3.0% (SE), 12.7 +/- 4.3%, and 14.6 +/- 6.7%, respectively. Three of nine moderately anaplastic astrocytomas had BrdU LIs of greater than 1% (average, 6.5 +/- 2.4%), whereas six had LIs of less than 1%. In two juvenile pilocytic astrocytomas, which are considered slow-growing, the BrdU LIs were unexpectedly high, averaging 6.5 +/- 1.4%. Thus pediatric medulloblastomas, glioblastomas multiforme, highly anaplastic astrocytomas, and a minority of moderately anaplastic astrocytomas had high proliferative potentials, whereas most of the moderately anaplastic astrocytomas had low proliferative potentials. Although the number of cases in this study is still too small to yield statistically significant comparisons, the results indicate that some pediatric tumors have considerably higher LIs than histologically similar adult tumors studied previously. PMID- 3027609 TI - Regrowth patterns of supratentorial gliomas: estimation from computed tomographic scans. AB - To clarify the regrowth patterns of benign and malignant gliomas, we chose 27 intervals (between two operations or between an operation and autopsy) from 21 patients with pathologically verified recurrent supratentorial gliomas. Serial computed tomographic (CT) scans of these cases were analyzed to determine the doubling time (Td) calculated from the change in volume of enhanced and low density areas, the enhancement effect graded from 0 to 4 according to the Hounsfield number, and the presence of dissemination and contralateral extension. We studied 5 benign gliomas (including 1 case of radiation necrosis), 8 malignant astrocytomas, and 8 glioblastomas. The Td's of enhanced areas on CT scans of benign gliomas, malignant astrocytomas, and glioblastomas were 937 +/- 66.5 days, 65.1 +/- 29.4 days, and 48.1 +/- 20.9 days, respectively. The Td's of low density areas were 895 +/- 130.6 days, 70.8 +/- 22.2 days, and 50.5 +/- 14.7 days. There was a significant correlation between the Td's of the enhanced and low density areas (0.97). The enhancement effect increased at recurrence in 55% of the cases, with an average increase of 1.1 grades. The increase in enhancement effect at recurrence showed a tendency to become smaller as the tumor's degree of anaplasia increased. Radiotherapy was effective in significantly retarding the growth rate of malignant gliomas, whose Td's were doubled. Although the Td's of both enhanced and low density areas of benign gliomas were significantly longer than those of malignant gliomas, there was no significant difference in the Td's of enhanced areas between malignant astrocytomas and glioblastomas.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3027610 TI - Neurologic complications after gastric restriction surgery for morbid obesity. AB - We report the occurrence of neurologic complications in 23 patients who underwent gastric restriction surgery for the treatment of morbid obesity. Complications occurred 3 to 20 months after surgery. All the patients had had protracted vomiting for the first 3 months after the operation. The following syndromes were found: chronic or subacute symmetric polyneuropathy (12 patients), acute severe polyneuropathy (1 patient), burning feet syndrome (2 patients), meralgia paresthetica (3 patients), myotonic syndrome (1 patient), posterolateral myelopathy (2 patients), and Wernicke-Korsakoff encephalopathy (2 patients). The patients suffering from burning feet syndrome and those with Wernicke-Korsakoff encephalopathy showed a clear improvement after parenteral thiamine treatment. As to the rest of the patients, the occurrence of the complications seems to be linked to nutritional causes, although no such deficiencies were detected. PMID- 3027613 TI - Immunoglobulin deposits in peripheral nerve endings detected by skin biopsy in patients with IgM M proteins and neuropathy. AB - Immunofluorescence studies of sural nerve and skin biopsies from three patients with IgM M proteins and clinical neuropathy showed that IgM M protein was bound to the nerve myelin in two patients and by the peri- and endoneurium in one. It is suggested that immunohistochemical studies of skin biopsies provide a simple effective method of detecting immunoglobulin binding to peripheral nerves in patients suspected of having an autoimmune neuropathy. In contrast to sural nerve biopsy, skin biopsy does not cause sensory loss or pain in a denervated area and can easily be repeated. PMID- 3027612 TI - The effect of electrical stimulation of medial temporal lobe structures in epileptic patients upon ACTH, prolactin, and growth hormone. AB - We measured ACTH, cortisol, prolactin, and growth hormone concentrations in patients with temporal lobe epilepsy who were having depth electrode studies. During the collection period, electrical stimuli were applied to amygdala and hippocampus to establish after-discharge thresholds. After-discharges that lasted at least 10 seconds or seizures caused secretion of ACTH and prolactin but not growth hormone. Stimuli that did not produce after-discharges of this duration inhibited ACTH secretion, but had no effect on prolactin or growth hormone secretion. PMID- 3027611 TI - Synaptic vesicle abnormality in familial infantile myasthenia. AB - In familial infantile myasthenia (FIM) the miniature end-plate potential (MEPP) amplitude is normal in rested muscle, but stimulation in vitro at 10 Hz decreases it abnormally, and neuromuscular transmission fails in a few minutes. In search of a morphologic correlate of the transmission failure, we analyzed the densities and diameters of synaptic vesicles in deep and superficial regions of nerve terminals in external intercostal muscles of three FIM patients and three nonweak controls before and after 10-Hz stimulation for 10 minutes. The densities of superficial or deep synaptic vesicles before or after stimulation in FIM were not significantly different from the corresponding control values. The diameters of superficial and deep synaptic vesicles before stimulation were significantly smaller in the three FIM patients than in the three controls. Stimulation in the FIM patients reduced the MEPP amplitude by 51 to 75%, but increased the vesicle diameter in two patients and did not change the vesicle diameter in one patient. Stimulation in the controls reduced the MEPP amplitude by only 16 to 34%, decreased the vesicle diameter in two, and did not change the vesicle diameter in one. Stimulation after treatment with 1 mg/dl hemicholinium markedly reduced the MEPP amplitude in the controls, had no further effect on the transmission defect in FIM, and had no consistent effect on vesicle diameter in FIM or controls. We conclude that synaptic vesicles are abnormally small in rested muscle in FIM, but vesicle size cannot be reliably correlated with the MEPP amplitude in FIM or controls. PMID- 3027614 TI - Somatosensory evoked potentials in chronic acquired demyelinating peripheral neuropathy. AB - We recorded somatosensory evoked potentials (SEPs) over the scalp in eight patients with chronic acquired demyelinating peripheral neuropathy. They were obtained from 15 nerves in which sensory nerve action potentials (SNAPs) were absent or not more than 1 microV, but from which motor responses could be elicited. Motor and sensory (SEP-derived) conduction velocity was determined from the difference in response latency with wrist and elbow stimulation. In 11 nerves, afferent conduction velocity was slowed. In 10, there was relatively equal slowing in sensory and motor axons, whereas in 1 there was disproportionate slowing in afferent fibers. In four nerves, afferent conduction velocity was within the normal range despite slowing of motor conduction. We conclude that SEPs may be useful in evaluating peripheral sensory conduction in the absence of SNAPs, but can provide misleadingly normal data, presumably because of central amplification of an attenuated response arising from a few axons that conduct normally. PMID- 3027615 TI - Membranous glomerulonephritis associated with chronic progressive demyelinating neuropathy. AB - Glomerulonephritis is known to be associated with the Guillain-Barre syndrome. We report the first case of a chronic progressive demyelinative neuropathy with membranous glomerulonephritis. PMID- 3027617 TI - [Regaining operability in a case of extensive cancer of the breast]. PMID- 3027618 TI - [Conservative surgical therapy of breast carcinoma. Our experience]. PMID- 3027619 TI - The value of computed tomography to the staging of non-small-cell primary bronchogenic carcinoma: a prospective study. AB - The value of computed tomography (CT) for the diagnosis of non-small-cell primary bronchogenic carcinoma with regard to T and N classifications was prospectively evaluated in a series of 29 patients. The sensitivity of CT in evaluating the extension of tumor to pleura or mediastinum was 100%, with only a 76% specificity. Computed tomography demonstrated 73 lymph nodes greater than or equal to 10 mm and 55 lymph nodes less than 10 mm in 27 patients. Invasive staging showed 23 lymph nodes greater than or equal to 10 mm and 22 nodes less than 10 mm which were not visualized by CT. Malignant invasion was found at histology in only one of these lymph nodes. The majority of nodes not visualized by CT were localized in the left paratracheal group, right and left tracheobronchial groups and the aortopulmonary window. PMID- 3027620 TI - [Hypertension therapy in the elderly. Our experience with converting enzyme inhibitors]. AB - Arterial hypertension shows, in the elderly, particular features and special problems connected with its pharmacological treatment. In our work ten patients, aged between 65-75, suffering from essential hypertension, were examined for eight weeks. At the end of this period, we observed a significant reduction of systolic and diastolic pressure, heart rate being unchanged. We didn't observe any significant change in the metabolic parameters considered (uricemia, creatininemia, triglycerides and cholesterol). No patient had to interrupt the treatment as a consequence of side effects. According to our data, we can affirm that Captopril reduces arterial pressure gradually and doesn't cause orthostatic hypotension, being thus very useful in the elderly. PMID- 3027616 TI - [Variations of the angiotensin converting enzyme in the serum of patients with ARDS during mechanical ventilation. Comparison with a control group]. PMID- 3027622 TI - [Correlations between colposcopic and histologic findings]. PMID- 3027621 TI - [Cyto-histologic aspects of viral infections and relation to carcinoma in situ]. PMID- 3027623 TI - Decreased striatal opiate delta-receptors in the rat model of persistent dyskinesia induced by iminodipropionitrile. AB - Chronic administration of iminodipropionitrile (IDPN) causes a persistent behavioral syndrome which consists of hyperactivity, vertical neck dyskinesias and lateral head twitches. D-Ala-D-Leu-enkephalin binding revealed a 26% decrease in the number but no change in the affinity of delta-opiate receptors in the striata of IDPN-treated rats. These findings are similar to those seen in the brains of patients with Huntington's disease. Further studies are needed to clarify the relationship of these findings to the phenomenology of the IDPN induced dyskinetic abnormalities. PMID- 3027624 TI - D-baclofen does not antagonize the actions of L-baclofen on rat neocortical neurons in vitro. AB - The ability of D-baclofen to antagonize the actions of L-baclofen on rat neocortical neurons was investigated. Intracellular recordings were made from neurons in cortical layers 2 and 3 in an in vitro slice preparation. Baclofen stereoisomers were applied at known concentrations in the superfusion medium. At a concentration of 3 microM, L-baclofen produced approximately 70% depressions of excitatory and inhibitory postsynaptic potentials (EPSPs and IPSPs) that were evoked by stimulation of superficial cortical layers. L-baclofen also hyperpolarized neocortical neurons. These hyperpolarizations were accompanied by decreases in neuronal input resistance and in direct excitability. We have shown previously that these latter effects are secondary to the action of baclofen to increase the potassium conductance of neocortical neurons. D-baclofen, at concentrations of 1-100 microM, did not antagonize depressions by L-baclofen of EPSPs and IPSPs nor the action of L-baclofen to increase the potassium conductance of neocortical neurons. At concentrations of 50-100 microM, D baclofen produced 20-30% effects when applied alone, thus suggesting that these concentrations of D-baclofen produced a significant degree of receptor occupancy. Our results demonstrate that D-baclofen is not an antagonist or high affinity partial agonist at the receptors through which baclofen exerts its effects on single neurons in the rat neocortex. PMID- 3027625 TI - Effects of food deprivation and high fat diet on opioid receptor binding in rat brain. AB - The effect of food deprivation for 72 h or a high fat diet on [3H]naloxone binding in the discrete brain regions of male lean Zucker rats was studied. In the midbrain, both treatments increased Bmax for the high-affinity site with no change in Kd. In the cortex, the high fat diet increased Bmax for the high affinity site. These results suggest that dietary manipulations could produce significant changes in the endogenous opioid system. PMID- 3027626 TI - Evidence that endogenous enkephalins produce delta-opiate receptor mediated neuronal inhibitions in rat dorsal horn. AB - Kelatorphan, a full inhibitor of aminopeptidases, enkephalinase and dipeptidylaminopeptidase, enzymes which degrade the enkephalins, produced inhibitions (around 50%) of dorsal horn C fibre evoked responses in the rat, following local application. The inhibitions were reversed by the selective delta opiate receptor antagonists ICI 174,864. Furthermore the inhibitions produced by two doses of Tyr-D-Ala-Gly-MePhe-Gly-ol (DAGO), a potent selective mu-opiate receptor agonist, were not altered in the presence of kelatorphan, so demonstrating an additive effect of the mu-agonist and the enzyme inhibitor. The inhibitions produced by the enzyme inhibitor would seem due to an increased availability of endogenous opioids, presumably the enkephalins which act via the delta-opiate receptor. PMID- 3027627 TI - Tifluadom, a kappa-opiate agonist, acts as a peripheral cholecystokinin receptor antagonist. AB - Tifluadom, a benzodiazepine kappa-opiate agonist, stereoselectively inhibited the binding of 125I-CCK to pancreatic membranes (IC50 = 47 nM). Several other opiate agonists were ineffective. Scatchard analysis indicated the inhibition of CCK binding by tifluadom was competitive in nature. Tifluadom (1 microM) did not displace 125I-CCK binding to brain tissue or 125I-gastrin binding to fundic glands. In the isolated guinea pig gallbladder, tifluadom antagonized CCK-8 induced contractions with an estimated pA2 of 6.8. These data demonstrate that tifluadom is a peripherally selective CCK antagonist. This unique action could contribute to its reported analgesic and appetite stimulatory properties. PMID- 3027629 TI - Hepatocellular carcinoma in the Gisborne district. PMID- 3027630 TI - Cough with angiotensin converting-enzyme inhibitors. AB - Since there are isolated case reports linking cough with angiotensin converting enzyme (ACE) inhibitor treatment, we reviewed the case notes of patients attending a hypertension outpatient clinic. Of 126 patients, 37 were on medications other than ACE inhibitors, and none complained of cough. In contrast, 12 of 89 patients receiving an ACE inhibitor had noted cough. The symptoms remained when one ACE inhibitor was substituted for another, but disappeared when the drug was withdrawn. Cough was sufficiently irritating to require cessation of treatment in two patients. We conclude that cough is not uncommon during treatment with ACE inhibitors. PMID- 3027628 TI - Models for quick activation of transducin and spontaneous deactivation of activated rhodopsin in rod outer segments. AB - Serial-parallel activation (S-P) and parallel-parallel activation (P-P) models had been proposed to account for the quick activation of transmitter molecules (transducin) in the visual transduction process in rod outer segments. In these models, unbleached rhodopsin molecules are activated by bleached rhodopsin molecules, and both bleached and activated rhodopsin molecules activate transducin. This activation mechanism enables bleached rhodopsin molecules to transfer the information of photon detection quickly to transducin. In the previous models, however, the rhodopsin photoisomerization process and the deactivation process of activated rhodopsin molecules were excluded. In the present study, these two processes are incorporated into the models. The deactivation process of activated rhodopsin molecules is assumed to proceed spontaneously. It is presented that, in spite of the incorporation of these two processes, the present S-P and P-P models can account for the quick activation of transducin in rod outer segments. PMID- 3027631 TI - Glomus tumour of the arm: case report. AB - A patient with a glomus tumour of the arm had pain as the leading symptom and this was relieved by surgery. PMID- 3027632 TI - Crescentic glomerulonephritis developing in a patient taking enalapril. PMID- 3027633 TI - A comprehensive clinical validation of the nuclear stethoscope. AB - Five studies were conducted to examine the degree of variability to be expected during the use of the non-imaging nuclear probe (BIOS Inc.) under a variety of clinical conditions. Comparison of the ejection fraction (EF) readings between the nuclear probe and a gamma camera showed good agreement, with the nuclear probe tending to underestimate lower, and overestimate higher camera EF values [mean (S.D.) difference, 0.84% (6.06)]. A comparison of two nuclear probes showed a small mean (S.D.) difference of EF readings of 0.063% (2.26). EF readings obtained in normal subjects 6 weeks apart were reproducible and differed by a mean (S.D.) of 0.23% (4.42). The administration of placebo to 10 normal subjects followed by sequential measurements for 4 h produced EF changes large enough to mimic a clinical effect, the largest hourly change observed being 5.4%, indicating the need for strict placebo control in interventional experiments. Data on four patients with heart failure showed small non-significant EF changes in the 1 h after placebo administration but a wide intra-subject range of ejection time and time to peak filling measurements. This highlights the problem of accurate, reproducible cursor placement in such patients. The nuclear probe is a portable, low cost instrument which produces accurate EF measurements when compared with the gamma camera. PMID- 3027634 TI - 131I and 111In-labelled monoclonal antibody imaging of primary lung carcinoma. AB - Preliminary clinical studies have been carried out to determine whether the monoclonal antibody 791T/36 would localize in primary lung cancer to an extent sufficient for external detection by gamma scintigraphy. Radiolabelling of the antibody with 131I permitted visualization of three out of eight (38%) tumours using planar imaging with 99Tcm-labelled blood pool subtraction. Radiolabelling of the same antibody with 111In permitted visualization of tumour uptake in nine out of 13 (69%) tumours, without the need for image subtraction. Single photon emission computed tomography (SPECT) studies of seven patients demonstrated concentration of 111In-labelled antibody at the tumour site in each case, four of which were visualized on the planar images. The present study demonstrates localization of the 791T/36 antibody in primary lung carcinoma and confirms the superiority of 111In over 131I as a radiolabel for antibody imaging, especially when emission tomography is performed. These data indicate that further work will be required to determine whether this antibody will be a suitable carrier for cytotoxic agents in the therapy of lung cancer. PMID- 3027635 TI - Biologic course of cervical human papillomavirus infection. AB - To determine the natural course of cervical human papillomavirus infection, we prospectively studied all new patients referred to the colposcopy clinic at the Naval Hospital Bethesda, from April 1981 to August 1983, whose screening cervical cytology demonstrated features consistent with human papillomavirus infection as the only abnormality. Histologic confirmation of human papillomavirus infection was required for entry into the study. All patients were evaluated by repeat cytology, colposcopy, endocervical curettage, and colposcopically directed biopsy as indicated at intervals of three to six months. Patients who developed classic features of cervical intraepithelial neoplasia were treated by standard modalities, whereas patients with evidence of human papillomavirus infection without associated cervical intraepithelial neoplasia were not treated. Confirmation of the resolution of human papillomavirus infection required negative cytology and colposcopy on two consecutive evaluations. Of the 45 patients for whom complete follow-up data are available, five (11.1%) had cervical intraepithelial neoplasia at the time of their initial evaluation, 15 (33.3%) progressed to cervical intraepithelial neoplasia over an average of 10.9 months, 18 (40%) resolved over an average of 13.7 months, and seven (15.6%) persisted with neither progression nor resolution for an average of 21 months. These data suggest that about one-third of patients who have histologically confirmed human papillomavirus cervical infection can be expected to develop cervical intraepithelial neoplasia within a year. PMID- 3027636 TI - Bacterial protease-induced reduction of chorioamniotic membrane strength and elasticity. AB - We demonstrated that in vitro exposure to bacterial collagenase and collagenase producing microorganisms significantly reduces measures of strength (bursting load), elasticity, and work to rupture of human amniochorion in a dose-dependent fashion. An attenuated noncollagenase-producing stain did not alter these measures. These findings support previous suggestions that infection within the lower uterine segment adjacent to the cervix may mediate some instances of premature rupture of membranes. PMID- 3027637 TI - Varicella-zoster virus infections during pregnancy: hypothesis concerning the mechanisms of congenital malformations. AB - An analysis of the data on 52 infants whose mothers had contracted varicella during pregnancy revealed that 27 had congenital malformations ascribed to the maternal varicella infections, while another 25 developed herpes zoster in the early postnatal period. Most of the mothers whose infants had congenital malformations had contracted varicella within the first 20 weeks of gestation, whereas most of the mothers whose infants developed herpes zoster had varicella after 21 weeks of gestation. The clinical features of the various congenital malformations and dysfunctions after maternal varicella were diverse; however, there were peculiar segmental manifestations of anomalies of the skin, the peripheral nervous, the autonomic nervous, and the musculoskeletal systems, all of which receive common innervations from the same levels of the spinal cord. Most other dysfunctions may be ascribed to an encephalitis. Therefore, the mechanism of congenital malformations caused by varicella-zoster virus seems to be due not to fetal varicella but to the development of herpes zoster in utero and to an encephalitis associated with herpes zoster. At least one infant had congenital malformations that were due to maternal herpes zoster infection. PMID- 3027638 TI - [Radiotherapy in adenoid cystic carcinoma]. AB - Between 1962 and 1982, 45 patients with adenoid cystic carcinomas (ACC) of various localizations received a radiotherapy-treatment. Most of the cases had advanced tumors or recurrences. 41 patients underwent radiotherapy either alone or following an operation, in 35 cases with curative intention. 6 patients had already distant metastases at the beginning of treatment. In 4 cases only a palliative treatment of metastases of the skeleton and the lung was performed. Median two- and five-year survival of 35 patients with localized tumors was 83% and 45%, respectively (29/35 two-years and 14/31 five years). Best results were obtained by operation and postoperative radiotherapy (10/18 = 55.5% five year survival). Radiotherapy alone resulted in a five-year survival of only 37.5% (3/8). Advanced ACC or postoperative recurrences should be treated by surgery as radically as possible, followed by a radiotherapy with generously sized portals and doses between 60 to 65 Gy. Radiotherapy alone leads to comparatively poor results. Large and inoperable tumors may well be treated by a combination of hyperthermia and radiotherapy. PMID- 3027639 TI - [Analysis of the effect of selection in a therapy study]. AB - 282 patients with SCLC were treated for the first time at the Chest-Hospital Heidelberg-Rohrbach from July 1981 until November 1983. Only 85 were integrated into a German randomized multicenter study of combined chemo- and radiotherapy, the other 197 were treated with individual therapeutic regimens. Generally, randomized and non-randomized patients were similar with respect to the prognostic factors and survival, but the group of excluded patients was more heterogeneous. Further analysis revealed prognostic relevance to surgical treatment, emergency treatment, various contra-indications and a poor general condition, but patients older than 70 years should not be generally excluded. General implications for clinical trials are that the clinical and methodological relevance of each criterion of exclusion should be critically examined and the baseline data on all excluded patients collected prospectively. PMID- 3027641 TI - Possible role of galactose-1-P-uridyl transferase activity deficiency in red blood cells in the development of the presenile and senile cataract. AB - The activity of red blood cells galactose-1-P-uridyl transferase in 64 patients with presenile and senile cataracts (nondiabetics) and in 41 age-matched controls was investigated. All control subjects examined have shown normal enzymatic levels, while 21.9% of patients with presenile cataracts and 21.7% of patients with senile cataracts presented a statistically significant reduced enzymatic activity (mean +/- 2 SD in controls). PMID- 3027643 TI - Multiple opioid systems and pain. PMID- 3027642 TI - Antiviral therapy with ganciclovir for cytomegalovirus retinitis and bilateral exudative retinal detachments in an immunocompromised child. AB - A child with bilateral cytomegalovirus (CMV) retinitis, vitritis, and exudative retinal detachments, who was in remission from stage IV neuroblastoma and status post-chemotherapy and autologous bone marrow transplantation, was treated with ganciclovir. The drug is a new acyclic nucleoside antiviral drug with potent antiCMV activity. There was bilateral retinal reattachment, clearing of vitritis and regression of retinal exudates and hemorrhages, with concomitant resolution of viral shedding in urine and blood, on ganciclovir 7.5 mg/kg per day. There was recurrence of exudative detachments, vitritis and retinitis when the dose was reduced to 2.5 mg/kg per day, and regression of these findings when the dose was again increased to 7.5 mg/kg per day. Despite continued therapy at this dose, a relapse occurred. When the dose of drug was doubled to 15 mg/kg per day, there initially was a partial therapeutic response, followed by a subsequent relapse. No further response was seen when the dose was increased to 19.5 mg/kg per day. This patient was treated with ganciclovir for a total of 192 days. No adverse reactions to ganciclovir were seen. On the last day of drug administration, there were persistent bilateral exudative retinal detachments and progressive optic nerve head involvement with optic disc pallor, despite quiescence of the retinitis. PMID- 3027640 TI - Studies on the standardization of mistletoe preparations. AB - The newly isolated phenylpropanoids syringin, syringenin-apiosylglucoside, eleutheroside E and the high molecular lectins and viscotoxins were selected for a standardization of mistletoe extracts and drug preparation. The phenylpropanoids found in all alcoholic and aqueous extracts were suitable for an HPLC fingerprint analysis, and for quantitative determination. The lectin content of the drug preparations was determined by single radial immunodiffusion. As shown by the isoelectric focussing method, Iscador and fermented mistletoe extracts contain only the mistletoe lectins ML II/III, whereas the proteins of the ML I complex are missing. For identification and quantitative determination of viscotoxins, an HPLC method was designed. PMID- 3027644 TI - [Papillomavirus: recent progress]. PMID- 3027645 TI - [Calcium-regulating hormone content of the blood in rickets in children]. PMID- 3027647 TI - [Clinical characteristics of serous meningitis due to Coxsackie B enteroviruses]. PMID- 3027646 TI - [Characteristics of cyclic nucleotide metabolism in vitamin D-deficient rickets]. PMID- 3027648 TI - [The renin-angiotensin system. Application in pediatrics. 1]. PMID- 3027649 TI - [Epstein-Barr virus encephalitis. Apropos of a case in a child]. AB - Encephalitic illnesses with Epstein-Barr virus (EBV) only represent a small percentage of the causes of viral encephalitis. Clinical symptoms are not specific. Biological standards tests carry few elements of direction, only serologic test of infectious mononucleosis confirms the diagnostic of recent EBV infection. Previous observation shows that the initial study can present similarities with herpes simplex encephalitis, which forces the immediate start of treatment by Acyclovir. PMID- 3027650 TI - [Fulminant herpesvirus hepatitis in a child. Apropos of 2 cases]. AB - The authors report two cases of fulminant hepatitis in children. Coagulopathy was early demonstrated in the two patients. The illness progressed rapidly and the two children died before receiving antiviral treatment. Herpes virus was demonstrated in the hepatocytes by electron microscopy. PMID- 3027651 TI - [The renin-angiotensin system. Applications in pediatrics. (II)]. PMID- 3027652 TI - [A study of a newly designed method in pharmacoangiography with a vasoconstrictor]. PMID- 3027655 TI - High speed chemical DNA sequence determination. PMID- 3027653 TI - Nucleotide sequence of Bacillus phage phi 29 genes 14 and 15: homology of gene 15 with other phage lysozymes. AB - The nucleotide sequence of Bacillus phage phi 29 genes 14 (g14) and 15 (g15) have been determined and shown to encode proteins with molecular weights of 15,014 and 28,022, respectively. The g14 open reading frame (ORF) was confirmed by sequencing a sus14(1241) mutant. Gene product 15 (gp15) has considerable homology with Salmonella phage P22 lysozyme and lesser homology with Escherichia coli phage T4 lysozyme. Putative translation signals are identified. In addition, the role of a previously described promoter, B2, is discussed. PMID- 3027656 TI - Purification and properties of a single strand-specific endonuclease from mouse cell mitochondria. AB - A nuclease was purified from mitochondria of the mouse plasmacytoma cell line, MCP-11 which acts on single-stranded DNA endonucleolytically and appears to have no activity upon native DNA. It degrades unordered RNA somewhat more effectively than it does DNA. The enzyme activity and the major detectable polypeptide migrate to a position corresponding to an Mr of 37,400 on denaturing polyacrylamide gels; in its native form the activity has an S value of 4.7, which corresponds to a molecular weight of roughly 73,000. The single-strand DNase activity has a pH optimum near 7.5, requires a divalent cation and is inhibited by EDTA, phosphate, KCl and NaCl. The enzyme is remarkably similar to fungal mitochondrial enzymes whose absence in various mutants correlates with defective DNA repair and recombination. It reacts weakly with antibody to a form of such an enzyme from Neurospora crassa. PMID- 3027654 TI - Molecular cloning of a steroid-regulated 108K heat shock protein gene from hen oviduct. AB - The natural gene for a steroid inducible 108K heat shock protein has been isolated from a lambda genomic library prepared from hen oviduct tissue. Genomic DNA blots indicate that it exists as a single copy gene in the chick oviduct haploid genome. The 9.9 kilobase gene codes for a messenger RNA of 2733bp (21) and is split into 18 exons as established by sequence comparison of cDNA and genomic clones. The 3' end of the gene contains a repetitive element which shares homology with the CR1 family of repeats. The first exon contains both the untranslated leader and coding regions of the gene. The promoter region is rich in G + C residues (70%) and the dinucleotide CG. This 5' flanking segment contains bases similar both in sequence and location to the Goldberg-Hogness TATA homology and consensus sequence CCAAT. A consensus sequence located upstream of steroid hormone responsive chicken genes is found at -267 and on a reverse orientation at -593. The structure of this gene is of interest since the presence of introns in heat shock genes is rare in any species examined to date. Furthermore, this gene lacks the previously described heat shock promoter consensus sequence (C-GAA-TTC-G) present in other species. PMID- 3027659 TI - Inhibition of restriction endonuclease Nci I cleavage by phosphorothioate groups and its application to oligonucleotide-directed mutagenesis. AB - M13 RF IV DNA where phosphorothioate groups are incorporated at restriction endonuclease Nci I recognition sites in the (-)strand is efficiently nicked by the action of this enzyme. Incubation of such nicked DNA with exonuclease III produces gapped DNA. The gap can be filled by reaction with deoxynucleoside triphosphates and DNA polymerase I. When this sequence of reactions is performed with DNA containing a mismatch oligonucleotide primer in the (-)-strand mutational frequencies of 70-90% can be obtained upon transformation. The general nature of this methodology has been further shown to be applicable to other restriction enzymes such as Hind II, Pst I and Fsp I. The mutational frequency obtained using these enzymes is between 40-80% mainly because of less efficient nicking and gapping. Studies on inhibition of Nci I cleavage show that in addition to a phosphorothioate group at the position of cleavage an additional group in the 5'-neighbouring position is necessary for complete inhibition. PMID- 3027658 TI - The ILV5 gene of Saccharomyces cerevisiae is highly expressed. AB - The nucleotide sequence of the yeast ILV5 gene, which codes for the branched chain amino acid biosynthesis enzyme acetohydroxyacid reductoisomerase, has been determined. The ILV5 coding region is 1,185 nucleotides, corresponding to a polypeptide with a molecular weight of 44,280. Transcription of the ILV5 mRNA initiates at position -81 upstream from the ATG translation start codon and terminates between 218 and 222 bases downstream from the stop codon. Consensus sequences have been identified for initiation and termination of transcription, and for general control of amino acid biosynthesis, as well as repression by leucine. The ILV5 gene is regulated slightly by general amino acid control. Codon usage of the ILV5 gene has the strong bias observed in yeast genes that are highly expressed. In agreement with this, the reductoisomerase monomer, with an apparent molecular weight of 40,000, has been identified in an SDS polyacrylamide gel pattern of total soluble yeast proteins as a gene dosage dependent band. PMID- 3027657 TI - Molecular analysis of the interaction between an enhancer binding factor and its DNA target. AB - The fine contacts of a mouse nuclear factor, called PEB1, with the B enhancer of polyoma virus were analyzed. It protects against DNaseI attack a region of about 50 base pairs that can be divided in two domains. The first contains a GC-rich palindrome and the homology to the SV40 enhancer. The second is homologous to a sequence in the immunoglobulin (Ig) heavy chain gene enhancer. Methylation interference and protection experiments reveal strong specific contacts only with a purine rich track on the late coding strand of the early proximal part of the palindrome. Deletion analysis show that the minimal sequences necessary for binding include only the first domain. The Ig homology contributes only weakly to the binding. The minimal core is similar to the core of the B enhancer defined in vivo. The interactions we observe here are reminiscent of those of TFIIIA positive transcription factor and the 5SRNA gene of Xenopus. PMID- 3027660 TI - The miniF plasmid C protein: sequence, purification and DNA binding. AB - The C (pifC) protein of miniF represses transcription of its own gene by binding to the pif operator (pifO); it is also needed for replication initiated from the miniF primary origin (ori-1). We have determined the nucleotide sequence of the C gene. The gene has been inserted into an expression vector under Ptrp control where it is expressed at high levels. The C protein has been purified from cells carrying the Ptrp-C plasmid, and a preliminary study of C protein-DNA binding properties has been carried out. C protein binds strongly to pifO, and weakly to sequences in the ori-1 region. PMID- 3027662 TI - Tricircular mitochondrial genomes of Brassica and Raphanus: reversal of repeat configurations by inversion. AB - We constructed complete physical maps of the tripartite mitochondrial genomes of two Crucifers, Brassica nigra (black mustard) and Raphanus sativa (radish). Both genomes contain two copies of a direct repeat engaged in intragenomic recombination. The outcome of this recombination in black mustard is to interconvert a 231 kb master chromosome with two subgenomic circles of 135 kb and 96 kb. In radish, a 242 kb master chromosome interconverts with subgenomic circles of 139 kb and 103 kb. The recombination repeats are 7 kb in size in black mustard and 10 kb in radish, and are nearly identical except for two insertions in the radish repeat relative to the black mustard one. The two repeat configurations present on the master chromosome of black mustard are located on the subgenomes of radish and vice-versa. To explain this, we postulate the existence of an evolutionarily intermediate mitochondrial genome in which the recombination repeats were (are) present in an inverted orientation. The recombination repeats described for these two species are completely different from those previously found in the closely related species B. campestris, implying that such repeats are created and lost frequently in plant mitochondrial DNAs and making it less than likely that recombination occurs in a site-specific manner. PMID- 3027661 TI - D protein of miniF plasmid acts as a repressor of transcription and as a site specific resolvase. AB - Two activities of the D protein of the miniF plasmid have been found. Divergent promoters in ori-1 ("primary" replicative origin) of miniF are both repressed in cells which produce D protein. The mobilization of plasmids containing the ori-1 region by the F conjugation system is also repressed by D protein. In the former case D appears to act as a transcriptional repressor, whereas in the latter case D protein acts by resolving cointegrates of F and the mobilized plasmid. D protein resolves dimers whose monomer units contain the rfsF sequence needed for recA-independent, site-specific recombination of F. The nucleotide sequence of the D gene was determined. The D gene region contains two oppositely-oriented open reading frames which have the same reading phase and substantially overlap. Transposon insertion mutants were used to show that the gene for D protein occupies the top-strand (left-to-right) open reading frame. PMID- 3027663 TI - A novel chlorophyll a/b binding (Cab) protein gene from petunia which encodes the lower molecular weight Cab precursor protein. AB - The 16 petunia Cab genes which have been characterized are all closely related at the nucleotide sequence level and they encode Cab precursor polypeptides which are similar in sequence and length. Here we describe a novel petunia Cab gene which encodes a unique Cab precursor protein. This protein is a member of the smallest class of Cab precursor proteins for which no gene has previously been assigned in petunia or any other species. The features of this Cab precursor protein are that it is shorter by 2-3 amino acids than the formerly characterized Cab precursors, its transit peptide sequence is unrelated, and the mature polypeptide is significantly diverged at the functionally important N terminus from other petunia Cab proteins. Gene structure also discriminates this gene which is the only intron containing Cab gene in petunia genomic DNA. PMID- 3027664 TI - A bacterial protein requirement for the bacteriophage lambda terminase reaction. AB - The bacteriophage lambda terminase enzyme cleaves the cohesive-end sites of lambda DNA to yield the protruding 5'-termini of the mature molecule. In vitro, this endonucleolytic event requires a protein factor which has been isolated and purified from extracts of uninfected E. coli. The terminase host factor (THF) is a heat stable basic protein of M.W. approximately 22,000. The integration host factor (IHF) protein of E. coli can efficiently substitute for THF in the terminase reaction; however, THF can be demonstrated to be physically present in, and isolated with full biological activity from extracts of cells defective or deficient in IHF. PMID- 3027665 TI - Functional, developmentally expressed genes for mouse U1a and U1b snRNAs contain both conserved and non-conserved transcription signals. AB - Four genes that encode mouse U1a1, U1b2 and U1b6 snRNAs have been isolated from a mouse genomic DNA library. They all appear to be functional U1 genes since they are accurately transcribed into full length, capped snRNAs upon injection into Xenopus oocytes. A mouse pseudogene that is not transcribed in Xenopus oocytes was also isolated from the mouse genomic library. DNA sequence analysis of the 5' and 3' flanking regions of the functional genes revealed the presence of three highly conserved sequence elements that have been shown to be required for transcription initiation or 3' end formation in other U1 genes. Each of these U1 RNA genes also contains non-conserved sequences in the 5' flanking region that could function in their controlled expression during development. PMID- 3027666 TI - Methylated DNA-binding protein from human placenta recognizes specific methylated sites on several prokaryotic DNAs. AB - Methylated DNA-binding protein (MDBP) from human placenta recognizes specific DNA sequences containing 5-methylcytosine (m5C) residues. Comparisons of binding of various prokaryotic DNAs to MDBP indicate that m5CpG is present in the recognition sites for this protein but is only part of the recognition sequence. Specific binding to MDBP was observed for bacteriophage XP12 DNA, which naturally contains approximately 1/3 of its residues as m5C, and for Micrococcus luteus DNA, M13mp8 replicative form (RF) DNA, and pBR322 when these three DNAs were methylated at CpG sites by human DNA methyltransferase. Five DNA regions binding to MDBP have been localized by DNase I footprinting or restriction mapping in methylated pBR322 and M13mp8 RF DNAs. A comparison of their sequences reveals a common 5'-m5CGRm5CG-3' element or closely related sequence in which one of the m5C residues may be replaced by a T. In addition to this motif, one upstream and one downstream m5CpG as well as other common residues over an approximately 20-bp long region may be recognized by MDBP. PMID- 3027667 TI - Characterization of bone PG II cDNA and its relationship to PG II mRNA from other connective tissues. AB - Two cDNA clones encoding the small proteoglycan II (PG II) of bone were isolated from a lambda gt11 expression library. These clones expressed recombinant protein which was cross-reactive with polyclonal and monoclonal antisera to PG II molecules from several connective tissues. The longest clone, lambda Pg 20 was studied in detail. The clone was shown to encode PG II by hybrid selected translation and immunoprecipitation. Northern analysis showed two species of the PG II message of approximately 1.4 and 1.8 kb. Substantial amounts of PG II message were found in bone, tendon, articular cartilage, skin, smooth muscle and cornea. Trace amounts of message were also detected in liver and brain. Radiolabeled bovine PG II cDNA hybridized to RNA from several other species including the human, rat and chicken. The level of PG II mRNA in chick embryonic fibroblasts was sensitive to transformation by Rous sarcoma virus. PMID- 3027668 TI - Alternative splicing of SV40 early pre-mRNA in vitro. AB - Simian virus 40 (SV40) early pre-mRNA is spliced using either of two alternative 5' splice sites and a common 3' splice site to produce two mRNAs that encode the T and t antigens. We have studied alternative splicing of SV40 early pre-mRNA in vitro using a HeLa cell nuclear extract. Synthetic SV40 early transcripts are processed to T and t antigen mRNAs in vitro. As in SV40-infected cells in vivo, cleavage at the T antigen 5' splice site is more efficient than cleavage at the t antigen 5' splice site in vitro, although both of these 5' splice sites are utilized relatively inefficiently in vitro. The ratio of cleavage at the T and t antigen 5' splice sites is not changed significantly by a number of alterations in the conditions under which the in vitro splicing reactions are carried out. PMID- 3027670 TI - Studies towards the development of chemically synthesized non-radioactive biotinylated nucleic acid hybridization probes. AB - Non-radioactive nucleic acid hybridization probes have been constructed in which the reporter group is long chain biotin chemically linked to a basic macromolecule (histone H1, cytochrome C or polyethyleneimine). The modified basic macromolecule which carries many biotin residues can, in turn, be covalently linked to nucleic acids (DNA) via the bifunctional cross-linking reagents, glutaraldehyde, 1,2,7,8-diepoxyoctane, bis (succinimidyl) suberate or bis (sulfonosuccinimidyl) suberate. This provides a very sensitive probe by which as little as between 10-50fg of target DNA can be visualized using dot-blot hybridization procedures in conjunction with avidin or streptavidin enzyme conjugates. PMID- 3027671 TI - Nucleotide sequence and organization of dnaB gene and neighbouring genes on the Bacillus subtilis chromosome. AB - A region of the Bacillus subtilis chromosome containing dnaB, a gene essential for the initiation of chromosomal replication in B. subtilis, was cloned and nucleotide sequence of some 5000bp determined. The region consists of 4 open reading frames (ORFs) possibly comprising a single transcriptional unit. Two dnaB mutations, dnaB27 and dnaB19 were located within the first ORF which would give rise to a protein of 472 amino acids. The dnaB27 mutation involves codon at the position of 122 (replacement of Asp by Asn) close to a DNA binding domain and the dnaB19 codon 379 (replacement of Ala by Thr) close to a region rich in charged amino acids which may be alpha helical. The third ORF in the same transcriptional unit would produce a hydrophobic protein which might be involved in the DNA membrane binding function of dnaB gene. No homologous Escherichia coli genes have been found. PMID- 3027669 TI - Nucleotide sequence of the virulence gene virG of the Agrobacterium tumefaciens octopine Ti plasmid: significant homology between virG and the regulatory genes ompR, phoB and dye of E. coli. AB - The entire nucleotide sequence of the virG locus, from the octopine Ti plasmid of Agrobacterium tumefaciens strain 15955, has been determined. The virG gene is 801 nucleotides in length and has one open reading frame which encodes a protein of Mr 29,995. The virG gene is involved in the transcriptional activation of the Ti plasmid vir-loci, which occurs after induction by specific compounds present in plant exudate. Sequence analysis of the Agrobacterium virG protein showed significant homology with the Escherichia coli ompR, phoB and dye proteins, which are all positive regulatory genes for genes encoding envelope proteins. These results suggest that the virG gene encodes a positive regulatory protein which can activate vir gene expression. PMID- 3027672 TI - Immunoscintigraphy and radioimmunotherapy in patients with pancreatic carcinoma. AB - A new monoclonal antibody (BW 494/32) labeled with 131I or 111In was used for planar and tomographic immunoscintigraphy (IS) in patients with pancreatic carcinoma. It appears that IS for pancreatic carcinoma and its metastases remains a hopeful but still difficult procedure and labeling with 111In is of advantage and results in more convincing images in the case of tumor lesions distant from liver and spleen. Attempts at radioimmunotherapy with 131I-anti-CA 19-9 and with 131I-494/32 in a patient with local recurrence of a pancreatic cancer and with large liver metastases were without success because of extremely poor blood supply to the metastatic tumor masses. Intraarterial infusion of the tracer without or with blockade and perfusion of the common hepatic artery with saline solution could not enhance the tracer uptake compared to that after intravenous infusion. High intratumoral concentrations, however, as achieved e.g. by intratumoral instillation in animal studies, represent a necessary precondition for effective beta-irradiation of tumor lesions. PMID- 3027674 TI - More fibre makes sense. PMID- 3027673 TI - Spotlight on children. Burns and aftercare. PMID- 3027675 TI - Case-control study of dietary etiological factors: the Melbourne Colorectal Cancer Study. AB - As part of a large-scale investigation of colorectal cancer (CRC) incidence, etiology, and survival, a case-control study was conducted to identify dietary factors associated with the risk of CRC. The study compared 715 cases with 727 age- and sex-matched community controls. A quantitative diet history, assessed to be the most representative of the previous 20 years, was obtained from each subject and analyzed for both food groups and nutrients. The combination of a high-fiber and high-vegetable intake was found to be protective against large bowel cancer. Cruciferous vegetable intake was also found, although with less certainty, to be protective. Dietary vitamin C was protective for estimated intakes greater than 230 mg/day. Dietary Beta-carotene had no separate association with the risk of CRC. Beef intake was a risk factor in males but not in females. Fat intake was a risk factor for both males and females. A low intake of milk drinks was a risk for both males and females. A high intake of pork and fish was protective. The use of vitamin supplements was highly protective. A risk score, which was calculated as the number of risk factors an individual has in his or her diet, showed an increasing monotonic relationship with risk of CRC. The effects of the dietary variables were similar for colon and rectal cancer and, with the exception of beef, were similar for males and females. PMID- 3027676 TI - Nursing care of patients experiencing cisplatin-related peripheral neuropathy. PMID- 3027677 TI - [Results of chemotherapy of patients with lung cancer]. PMID- 3027678 TI - Nosebleed in children. Background and techniques to stop the flow. AB - Nosebleed in children can result from dryness and picking of the resultant crust over the anterior part of the nasal septum, trauma to the nose, juvenile angiofibroma, or disorders of hemostatic mechanisms. In most cases it is not difficult to treat; often the primary care physician can assist a patient by giving instructions over the telephone to a parent. In the office or hospital, the usual measures are firm pressure, placement of a piece of cotton dipped in a cocaine-epinephrine solution, taking of a brief history, application of petrolatum, and taping of the nose. If bleeding persists, anterior nasal packing and, rarely, posterior packing should be performed. Maxillary artery ligation is done in cases of severe epistaxis. Special care must be taken with children who have a bleeding disorder or who are recovering from adenoidectomy. PMID- 3027679 TI - Laboratory testing for infectious mononucleosis. Cautions to observe in interpreting results. AB - In the vast majority of cases, diagnosis of infectious mononucleosis is relatively simple and the illness is not serious. Performing tests for specific Epstein-Barr virus (EBV) antibodies in these cases is not necessary. However, when the clinical manifestations are atypical or unusually severe, especially when the heterophil antibody test is negative, specific EBV antibody tests may be needed. The EBV antibodies used in diagnosis are IgG antibodies to viral capsid antigen (VCA), IgM antibodies to VCA, and antibodies to early antigen (anti-D) and Epstein-Barr nuclear antigen (EBNA). The diagnosis of infectious mononucleosis may be made when IgG-VCA, IGM-VCA, and anti-D antibodies are present and EBNA antibodies are absent. EBNA antibodies appear later and, together with IgG-VCA antibodies, persist indefinitely. Still, infectious mononucleosis often cannot be diagnosed with certainty because of the difficulties in interpreting laboratory findings. The diagnosis must be made with caution and possible sources of error considered when test results are interpreted. PMID- 3027680 TI - Neurologic complications of human immunodeficiency virus infection. What diagnostic features to look for. PMID- 3027681 TI - Advances in endocrine hypertension. PMID- 3027682 TI - Role of reactive oxygen species and free radicals from melanins in photoinduced cutaneous inflammations. PMID- 3027684 TI - [Pathology of the digestive tract in AIDS]. PMID- 3027683 TI - Fetal intestinal microvilli in human amniotic fluid. AB - The intestinal microvilli of fetal origin in human amniotic fluid were purified by Ca2+ precipitation of contaminating organelles followed by differential centrifugation of microvillar membranes. In the purified preparation, the specific activity of the microvillar marker-enzymes maltase and sucrase increased about 77-fold over that in cell-free amniotic fluid. Significant contamination of the purified preparation by endoplasmic reticulum (microsomes) and lysosomes was ruled out on the basis of a low content of the marker enzymes glucose-6 phosphatase (microsomes) and acid phosphatase (lysosomes). Amniotic fluid microvilli contain typical enzymes of the fetal intestine including maltase, sucrase, trehalase, alkaline phosphatase and gamma-glutamyltransferase, and their morphology by electron microscopy resembles that of vesiculated intestinal microvilli. Prenatal detection of genetic diseases due to a deficiency of a protein expressed in these membranes or associated to abnormal microvilli seems feasible. PMID- 3027685 TI - [Immunocytochemical demonstration of actin in myothelial cells in proliferative diseases of the breast]. PMID- 3027686 TI - [Peripheral neuropathy in primary polycythemia]. PMID- 3027687 TI - [A case of LAV II-virus AIDS in Mali]. PMID- 3027689 TI - [Addison's disease in children. Etiology and physiopathologic acquisitions]. PMID- 3027688 TI - [Surgical treatment of hepatoma on a cirrhotic liver]. AB - From January 1, 1975 to July 1, 1984, 37 patients with a tentative diagnosis of hepatocarcinoma on cirrhosis were operated upon. There were 34 men and 3 women, aged from 32 to 82 years (mean: 60 years). The diagnosis of cirrhosis rested either on a history of liver failure associated with clinical and biochemical signs of hepatocellular dysfunction (4 cases), or on a positive liver biopsy (5 cases). In 20 cases the diagnosis was suspected on account of abnormal liver function tests, but in 8 patients it was revealed by macroscopic examination of the liver during surgery. Cirrhosis was attributed to chronic alcoholism in 25 cases, haemochromatosis in 11 cases and positive HBs antigen in 7 cases. The diagnosis of hepatocarcinoma, suggested by altered general condition or recent pain, rested on the finding of a tumoral image at scintigraphy (16 cases), ultrasonography (19 cases), computed tomography (7 cases), arteriography (17 cases) or laparoscopy (7 cases). In only 7/32 patients was the alpha-foetoprotein level higher than 500 ng/l. Surgery confirmed the diagnosis of hepatocarcinoma in every case but that of cirrhosis in only 14/37 cases; 10 patients had lesion of hepatic fibrosis and 1 had regenerative nodular hyperplasia. In 10 cases no accurate histological diagnosis could be made since the liver tissue sample had been taken too close to the tumour. The finding of normal liver tissue shows that one should not refrain from operating merely because the diagnosis of cirrhosis rests on clinical grounds. Since 1979, surgical treatment consists in an attempt to excise the tumour. Per-operative mortality is the same with excision surgery (21%) as with exploratory or palliative surgery (17%). Fourteen excisions were performed (i.e. aresectability rate of 38% for the series): 2 liver transplantations, 2 right hepatectomies, 2 left hepatectomies, 2 left lobectomies, 2 bisegmentectomies and 3 tumorectomies. The survival rate of 2 years was 27%, as opposed to 5% with exploratory or palliative surgery. Systematic monitoring of cirrhosis with ultrasonography should result in early diagnosis of hepatocarcinoma at a stage where limited hepatic excision is possible and the chances of surviving are highest. PMID- 3027691 TI - [Study of restrictase Sal GI biosynthesis and optimum conditions for its isolation]. AB - The effect of the growing phase of Streptomyces albus G. and the components of the culture medium on the biosynthesis of restriction endonuclease Sal GI was studied. The conditions of DNA sedimentation with streptomycin sulfate and polyethylenimine and separation of proteins with ammonium sulfate were investigated as well. A maximal quantity of the enzyme was observed in the cells in 18-20 h of cultivation in flasks on a shaker. The optimal concentrations of the medium components (g/l) were found by mathematical methods of the experiment planning: glucose--19,0; pepton--5.2; K2HPO4 X 3H2O - 5.0. The optimal concentrations for DNA sedimentation with streptomycin sulfate and polyethylenimine were found to be 1.8-2.0% and 0.2-0.3%, respectively. The optimal concentration of ammonium sulfate for protein separation is 70% of saturation. PMID- 3027690 TI - [Peripheral neuropathy in relation to LAV/HTLV III retrovirus infection. A clinical, anatomical and immunological study. 5 cases]. AB - An unexplained peripheral neuropathy was observed in five patients with positive serology for LAV/HTLV III. Three of them presented with polyneuropathy, one with chronic meningitis and oculomotor palsies, and one with a mononeuropathy. CSF was abnormal in 5/5, with elevated protein content (0.4-4 g/l) and abnormal cell count (29-65/mm3). Intrathecal production of LAV-specific IgG was demonstrated in 3/4 cases. Electromyographic examination showed reduced nerve conduction velocity in 4/5. Neuromuscular biopsy revealed microvasculitis with mononuclear cell infiltrates in 3/4 cases; characterization of these cells showed that they were predominantly non monoclonal T8 lymphocytes. Other symptoms of "AIDS-related complex" were present in all five patients. None had other causes of peripheral neuropathy. Thus, peripheral neuropathy can be the initial manifestation of LAV/HTLV III infection. Isolation of the virus from the nerve in one published case, and arguments for intrathecal synthesis of LAV-specific IgG suggest the direct role of this agent; however, the lymphocytic infiltration seen in three of our cases favours an indirect immune mechanism, as in other organs, such as lungs and lymph nodes. PMID- 3027692 TI - [Similarity and differences of the pathogenesis of Itsenko-Cushing syndrome variants]. AB - Proceeding from a study of the results of the investigation of 152 patients with Icenko-Cushing's syndrome the authors have arrived at the conclusion that hyperplasia, clear cell adenoma of monomorphous structure, dark cell and mixed adenomas of polymorphous structure, cancer of the adrenal cortex are different stages of a common pathological process. Such formation of morphological changes in the adrenal gland is one of the ways of development of Icenko-Cushing's syndrome. To confirm the proposed concept of the pathogenesis of this disease the authors provide some evidence of hyperplasia or the normal structure of nontumorous adrenocortical tissue in adenoma and cancer, of the absence of difference in a rise of the mean ACTH level in the blood in hyperplasia and different adrenocortical tumors. An increase in ACTH production in most of the patients should be regarded as one of the primary factors of the above morphological changes. The absence of suppression of 17-oxycorticosteroid excretion with urine in a test with 8 mg of dexamethasone in most of the patients with dark cell and mixed adenoma of polymorphous structure and adrenocortical cancer suggests a possibility of the development of certain autonomy of these neoplasms. PMID- 3027693 TI - [The role of heparin in the realization of the effect of various protein hormones]. AB - In immature female rats the ovarian mass increased by 74.3% 48 h after the administration of 50 units of LH. The stimulating LH effect on the ovary did not show itself against a background of the binding of circulatory heparin by multiple administration of protamine-sulphate (P-S) for 3 days. The blockade could be abolished by compensatory administration of heparin (twice at a dose of 50 units/100 g. Likewise, the stimulating ACTH effect on corticosterone secretion in adult male rats did not show itself in P-S-heparin binding. If in the blockade of discharge of the animal own ACTH by dexamethasone 2.5 units of exogenous ACTH resulted in an increase in corticosterone concentration from 9 to 16.2 Mg% in 15 min, in P-S single administration 5-10 min before ACTH, corticosterone concentration remained practically at the basal level. Heparin injection within the interval between P-S and ACTH administration ensured the manifestation of the stimulating ACTH effect, and blood corticosterone concentration increased in the same degree as in the administration of ACTH only. Thus, it can be assumed that the presence of reactive heparin in the blood is necessary for the implementation of action of some protein hormones. PMID- 3027694 TI - Nucleotide sequence determining the first cleavage site in the processing of mouse precursor rRNA. AB - The first step in the processing of 47S precursor rRNA in mouse cells is reproduced in vitro in an S-100 transcription reaction and consists of an endonucleolytic cleavage at residue +650 of the primary transcript followed by rapid degradation of the fragment upstream from residue +650. An analogous processing occurs in human rRNA. The mouse and human rRNA sequences are approximately equal to 80% conserved for 200 nucleotides on the 3' side of these processing sites, suggesting that this conserved region may be important in specifying the processing. To test this hypothesis, we constructed a systematic series of deletion mutants approaching the mouse rDNA processing region from both the 5' and 3' directions and analyzed the processing of their transcripts in vitro. The 5' boundary of the region required for processing is quite sharp and corresponds to the rRNA cleavage site at the 5' end of the conserved sequence region. The 3' boundary is more complex: The 3' deletions extending to between 250 and 130 nucleotides beyond the processing site cause about a 50% decrease in the amount of the processed RNA. A 3' deletion that extends to 109 nucleotides beyond the processing site greatly reduces the processing efficiency. Deletions to or beyond 91 nucleotides on the 3' side of the processing site virtually eliminate processing. Under altered ionic conditions, transcripts of 3' deletions extending to only 41 nucleotides beyond the processing site can still direct a low level of accurate processing. These results demonstrate that the mouse/human conserved sequence just on the 3' side of the primary rRNA processing site consists of several domains that direct and/or augment both the initial endonucleolytic cleavage and the closely coupled selective degradation of the upstream fragment that together constitute the primary rRNA processing event. PMID- 3027696 TI - Specific antiviral activity of a poly(L-lysine)-conjugated oligodeoxyribonucleotide sequence complementary to vesicular stomatitis virus N protein mRNA initiation site. AB - Antisense oligonucleotides represent an interesting tool for selective inhibition of gene expression, but their efficient introduction within intact cells proved to be difficult to realize. As a step toward this goal, small (13- or 15-mer) synthetic oligodeoxyribonucleotides have been coupled at their 3' ends to epsilon amino groups of lysine residues of poly(L-lysine) (Mr, 14,000). A 15-mer oligonucleotide-poly(L-lysine) conjugate complementary to the initiation region of vesicular stomatitis virus (VSV) N-protein mRNA specifically inhibits the synthesis of VSV proteins and exerts an antiviral activity against VSV when added in the cell culture medium at doses as low as 100 nM. Neither synthesis of cellular proteins nor multiplication of encephalomyocarditis virus was affected significantly by this oligonucleotide conjugate. The data suggest that oligonucleotide-poly(L-lysine) conjugates might become effective for studies on gene expression regulation and for antiviral chemotherapy. PMID- 3027695 TI - cDNA and derived amino acid sequence of ethanol-inducible rabbit liver cytochrome P-450 isozyme 3a (P-450ALC). AB - Administration of ethanol to rabbits is known to induce a unique liver microsomal cytochrome P-450, termed isozyme 3a or P-450ALC, which is responsible for the increased oxidation of ethanol and other alcohols and the activation of toxic or carcinogenic compounds such as acetaminophen and N-nitrosodimethylamine. To further characterize this cytochrome P-450 we have identified cDNA clones to isozyme 3a by immunoscreening, DNA hybridization, and hybridization-selection. The cDNA sequence determined from two overlapping clones contains an open reading frame of 1416 nucleotides, and the first 25 amino acids of this reading frame correspond to residues 21-45 of cytochrome P-450 3a. The complete polypeptide, including residues 1 to 20, contains 492 amino acids and has a molecular weight of 56,820. Cytochrome P-450 3a is approximately 55% identical in sequence to P 450 isozymes 1 and 3b and 48% identical to isozyme 2. Hybridization of clone p3a 2 to electrophoretically fractionated rabbit liver poly(A)+ RNA revealed multiple bands, but, with a probe derived from the 3' nontranslated portion of this cDNA, only a 1.9-kilobase band was observed. Treatment of rabbits with imidazole, which increases the content of isozyme 3a, resulted in a transient increase in form 3a mRNA, but this was judged to be insufficient to account for the known 4.5-fold increase in form 3a protein. Genomic DNA analysis indicated that the cytochrome P 450 3a gene does not belong to a large subfamily. PMID- 3027697 TI - Nucleotide sequence of Bacillus subtilis dnaB: a gene essential for DNA replication initiation and membrane attachment. AB - The complete nucleotide sequence of the Bacillus subtilis dnaB gene and its flanking regions was determined. The dnaB gene is essential for both replication initiation and membrane attachment of the origin region of the chromosome and plasmid pUB110. It has been known that there are two different classes (dnaBI and dnaBII) in the dnaB mutants; dnaBI is essential for both chromosome and pUB110 replication, whereas dnaBII is necessary only for chromosome replication. The nucleotide sequence revealed that dnaBI and dnaBII are two functional domains in the single dnaB gene. The mutation sites of two mutants, belonging to dnaBI and dnaBII, respectively, were also determined as substitutions of amino acids. The putative DnaB protein deduced from nucleotide sequence consists of 472 amino acids (55 kDa) with no cysteine residue. A 55-kDa polypeptide produced in an in vitro transcription-translation system was labeled with [35S]methionine but not with [35S]cysteine. The DnaB protein has a highly hydrophobic sequence of 20 amino acids in its N-terminal region, a possible DNA binding site, and two possible ATP binding sites. The dnaBI domain is between the DNA binding site and one of the ATP binding sites; the dnaBII domain is close to the other ATP binding site. Comparison of the amino acid sequence between the "dnaB protein" and those of other dna genes of Escherichia coli showed no homology, suggesting that the dnaB gene of B. subtilis may be analogous to a hitherto undiscovered gene in E. coli. PMID- 3027698 TI - Amino acid sequence of S-adenosyl-L-homocysteine hydrolase from rat liver as derived from the cDNA sequence. AB - Rat liver cDNA libraries constructed in lambda gt11 were screened for reactivity with polyclonal antibodies to native S-adenosyl-L-homocysteine (AdoHcy) hydrolase (adenosylhomocysteinase; EC 3.3.1.1). Five clones were isolated and sequenced. The amino acid sequence, deduced from the cDNA sequence, contained the sequence of eight peptides obtained by tryptic and cyanogen bromide fragmentation of rat liver AdoHcy hydrolase. Identification of the amino- and carboxyl-terminal peptides in the amino acid sequence showed that the complete sequence was obtained. A "fingerprint" sequence was found that is characteristic of dinucleotide-binding domains of many proteins. For AdoHcy hydrolase, this region from the lysine at position 213 to the aspartate at position 244, containing the sequence Gly-Xaa-Gly-Xaa-Xaa-Gly at positions 219-224, is presumably the site of binding for NAD+, which is required for the activity of the enzyme. PMID- 3027699 TI - Incorporation of a retinal rod cGMP-dependent conductance into planar bilayers. AB - The light-modulated current of vertebrate retinal rods flows through a 3',5' cyclic GMP-dependent conductance located in the outer segment plasma membrane. We report the incorporation into planar bilayers of a conductance derived from vertebrate rod outer segment membranes specifically activated by cGMP but not by cAMP, 5'-GMP, GTP, or 5'-AMP. When the mean currents were measured as a function of increasing cGMP concentration, maximal activation occurred at concentrations less than 50 microM. Washout of cGMP rapidly reversed the effect. The apparent half-saturating concentrations were between 12 and 27 microM. Sodium, lithium, cesium, and potassium supported current in the presence of low concentrations of Ca2+, Mg2+, and 100 microM cGMP; choline did not. Removal of the divalent cations reversibly increased the currents. When calcium was the only current-carrying cation, attenuated currents were seen. These experiments support the hypothesis that calcium is a permeant blocker of the conductance. At low concentrations of cGMP in solutions also containing 0.5 mM EDTA, brief current spikes occurred with amplitudes from 0.5 to 4 pA at 50 mV. These spikes differed from the well defined, unitary conductance steps usually associated with the opening and closing of ion channels. Occasionally we saw longer-lasting channel-like events; however, amplitude histograms did not resolve discrete conductance levels. PMID- 3027700 TI - Efficient and stable expression of recombinant fibronectin polypeptides. AB - We describe retroviral expression vectors containing cDNAs encoding part of fibronectin preceded by the signal and "pro" sequences of parathyroid hormone. The recombinant retroviruses were used to generate NIH 3T3 cell lines stably producing functionally active fragments of fibronectin. The recombinant fibronectins (deminectins) are processed and secreted by the cells and form disulfide-bonded dimers with themselves and with endogenous fibronectin subunits. The fibronectin-deminectin heterodimers are incorporated into the extracellular matrix. We describe cell lines producing six variant forms of deminectin corresponding to variant forms of fibronectin produced by alternative splicing. In constructing fibronectin cDNAs encoding the six variant forms, we also made use of the ability of retroviral vectors to generate cDNAs by accurate splicing of cloned genomic segments. These constructs should be valuable in analyses of the structure-function relationships of fibronectins. PMID- 3027701 TI - Phosphorylation of a high molecular weight DNA polymerase alpha. AB - Anti-human DNA polymerase alpha murine IgG SJK-287-38 [Tanaka, S., Hu, S.-Z., Wang, T. S.-F. & Korn, D. (1982) J. Biol. Chem. 257, 8386-8390] neutralized DNA polymerase alpha activity from rat embryonic fibroblasts infected with a temperature-sensitive transformation mutant of Rous sarcoma virus (tsLA24). After centrifugation of a crude cytosol fraction from log-phase cells in a 5-20% linear sucrose gradient, polypeptides of Mr approximately equal to 185,000 and 220,000 were immunoprecipitated only from gradient fractions containing DNA polymerase alpha activity. When similar cultures were incubated in medium containing [32P]orthophosphate, it was found that the Mr 220,000 protein was phosphorylated but that the other peptides specific for polymerase alpha activity did not contain detectable amounts of phosphate. Phospho amino acid analysis of the high molecular weight immunoprecipitable proteins indicated that the labeled amino acid was phosphoserine. Incubation of 2.5 units of crude DNA polymerase alpha with 4 units of agarose-immobilized alkaline phosphatase resulted in a nearly complete inhibition of DNA polymerase alpha activity. Subsequent incubation of this preparation with 5 or 50 microM ATP, but not the nonhydrolyzable analog adenosine 5'-[gamma-thio]triphosphate, restored the in vitro DNA polymerizing activity. These results demonstrate that a high molecular weight DNA polymerase alpha (Mr approximately equal to 220,000) is phosphorylated in cultured cells and that this protein is a substrate for a serine kinase rather than the tyrosine specific protein kinase of Rous sarcoma virus. The results suggest that phosphorylation/dephosphorylation reactions modulate the activity of this polymerase. PMID- 3027702 TI - Nucleotide sequence and characterization of the 5' flanking region of the rat Ha ras protooncogene. AB - Thyrotropin and cAMP stimulate growth of FRTL5 rat thyroid cells and increase c Ha-ras mRNA levels. To study the mechanism by which thyrotropin enhances c-Ha-ras expression in the thyroid, we constructed a genomic library of FRTL5 DNA in the bacteriophage vector EMBL3. Using a v-Ha-ras probe (0.7-kilobase Sst I-Pst I fragment), we isolated eight clones containing portions of the FRTL5 c-Ha-ras gene. Restriction mapping of one of these clones revealed a structure very similar to that previously reported for the rat c-Ha-ras gene. We determined the nucleotide sequence of exon 1 as well as 1.15 kilobases upstream from exon 1. Blot-hybridization analysis of FRTL5 thyroid cell mRNA was performed with three DNA fragments upstream of exon 1. Two of these probes were Pst I-Pst I fragments 0.4 and 0.55 kilobases long, 1.15 and 0.6 kilobases upstream of exon 1, respectively. The third probe, a 0.6-kilobase Pst I-HindIII fragment, was immediately upstream of exon 1 and included 54 base pairs of the 5' end of exon 1. The data revealed an upstream ("-1") exon, consistent with the homology between the nucleotide sequences of our clone and the human c-Ha-ras gene in this region. Primer extension of a synthetic 30-mer oligonucleotide probe complementary to exon +1 on a FRTL5 mRNA template revealed three transcription start (cap) sites, 182, 169, and 153 bases upstream of the ATG codon. "CCAAT boxes" are present 65 and 100 bases upstream from the initial cap site. Three "GC boxes" and two C + G-rich inverted repeats characteristic of the binding site for the transcription regulation factor Sp1 are present in the 177 base pairs upstream of the initial cap site. Comparison of the rat and human c-Ha-ras -1 exons showed an area of poor homology immediately upstream of the human cap sites, followed further upstream by a region of close homology (approximately equal to 60 base pairs). The nucleotide sequence of the rat -1 exon is more similar to the v-ras sequence than is the human. The v-ras gene is truncated relative to both human and rat -1 exons. PMID- 3027703 TI - Molecular mapping of point mutations in the period gene that stop or speed up biological clocks in Drosophila melanogaster. AB - The pero1 and the pers mutations in Drosophila melanogaster, which seem to eliminate or speed up, respectively, the clocks underlying biological rhythmicity, were mapped to single nucleotides. Chimeric DNA fragments consisting of well-defined wild-type plus mutant DNA subsegments were constructed, introduced into flies by germ-line transformation, and assayed for biological activity. These experiments localized both pero1 and pers to a 1.7-kilobase DNA fragment that is mostly coding DNA. Sequencing of this subsegment from each mutant showed that pero1 is completely accounted for by a nonsense mutation in the third coding exon of a 4.5-kilobase RNA transcribed from this locus. The pers mutation is also a single nucleotide substitution, in the fourth coding exon, which results in a serine-to-asparagine substitution in the per gene protein product. The functional significance of these changes is discussed with reference to the phenotypes of the two mutations. PMID- 3027704 TI - Modulation of gene expression in multiple hematopoietic cell lineages following retroviral vector gene transfer. AB - Retrovirus vectors offer a simple and highly efficient method for introducing new genes into mammalian cells. Here, we have examined the efficiency of gene transfer into hematopoietic cells with retrovirus vectors carrying the neomycin (neo) resistance gene expressed from different transcriptional regulatory regions. Direct infection of mouse bone marrow cells resulted in high efficiencies of gene transfer into a variety of myeloid progenitor cells, including pluripotent, erythroid, and granulocyte-macrophage colony-forming cells with all the vectors examined. However, the progeny derived from individual pluripotent progenitor cells infected with different vectors differed markedly in the proportion of G418-resistant progenitor cells, as judged by their ability to survive selection in the drug G418. This biological assay suggests that the highest level of expression was observed when the neo gene was expressed from constructs that contained the herpes thymidine kinase promoter rather than the viral long terminal repeat or the simian virus 40 early region promoter. In contrast, neo gene expression was highest in fibroblasts infected with the vector containing the simian virus 40 early region promoter. These results show that high and sustainable levels of gene expression in hematopoietic cells can be obtained with retrovirus vectors containing appropriate transcriptional regulatory regions. PMID- 3027706 TI - Epstein-Barr virus-containing B-cell line produces an interleukin 1 that it uses as a growth factor. AB - We report the establishment of a spontaneous interleukin 1 (IL-1)-producing subclone derived from the human Epstein-Barr virus (EBV)-containing B lymphoblastoid cell line (721 LCL) and show that the IL-1 produced by this B-cell subclone is distinct from other types of IL-1. The parental cell line 84.5, a deletion mutant of the 721 LCL cell line, can be induced to produce IL-1 activity when stimulated by certain inducers such as phorbol 12-myristate 13-acetate in the presence of fetal calf serum. From this parental 721/84.5 clone, a subclone, termed 3B6, has been developed. This 3B6 subclone has an immature B-cell phenotype, expresses only HLA class II DP subregion antigens, and spontaneously releases IL-1 in the culture supernatant with relatively few inhibitory molecules under serum-free culture conditions. The 3B6-derived IL-1 was purified from 3B6 conditioned medium with a three-step procedure. The molecular weight of this IL-1 is 13,500, and the isoelectric point values are pH 4.9 and 5.1 without any component focusing near pH 7. The N-terminal amino acid sequence differs markedly from those reported for the two IL-1 species produced by monocytes. The purified material shares several biological properties with monocyte IL-1, since it could induce the proliferation of murine thymocytes, the production of interleukin 2 by phytohemagglutinin-stimulated cloned HSB2 T cells, and the proliferation of human fibroblasts. However, this IL-1 activity could not be blocked by polyclonal anti monocytic IL-1 antibodies, and, more importantly, it was not pyrogenic in rabbits. Finally, it promotes the growth of B-cell clones derived from the parental 721/84.5 lines in the absence of fetal calf serum, which suggests that it could act as an autocrine growth factor in this Epstein-Barr virus-transformed B-cell line. PMID- 3027705 TI - Distinctive transforming genes in x-ray-transformed mammalian cells. AB - DNAs from hamster embryo cells and mouse C3H/10T1/2 cells transformed in vitro by x-irradiation into malignant cells transmit the radiation transformation phenotype by producing transformed colonies (transfectants) in two mouse recipient lines, the NIH 3T3 and C3H/101/2 cells, and in a rat cell line, the Rat 2 cells. DNAs from unirradiated cells or irradiated and visibly untransformed cells do not produce transformed colonies. The transfectants grow in agar and form tumors in nude mice. Treatment of the DNAs with restriction endonucleases prior to transfection indicates that the same transforming gene (oncogene) is present in each of the transformed mouse cells and is the same in each of the transformed hamster cells. Southern blot analysis of 3T3 or Rat-2 transfectants carrying oncogenes from radiation-transformed C3H/10T1/2 or hamster cells indicates that the oncogenes responsible for the transformation of 3T3 cells are not the Ki-ras, Ha-ras, or N-ras genes, nor are they neu, trk, raf, abl, or fms, although quick blot analysis using 11 oncogene probes detected increased transcripts of c-abl and c-fms in the 3T3 transformants containing oncogenic sequences from the x-ray-transformed C3H/10T1/2 cells. The work demonstrates that DNAs from mammalian cells transformed into malignancy by direct exposure in vitro to radiation contain genetic sequences with detectable transforming activity in three recipient cell lines. The results provide evidence that DNA is the target of radiation carcinogenesis induced at a cellular level in vitro. The experiments indicate that malignant radiogenic transformation in vitro of hamster embryo and mouse C3H/10T1/2 cells involves the activation of unique non-ras transforming genes, which heretofore have not been described. PMID- 3027707 TI - Potentiating and suppressive IgE-binding factors are expressed by a single cloned gene. AB - We have investigated expression of an IgE-binding factor (IgE-BF) cDNA in both COS-7 monkey kidney cells and Chinese hamster ovary cells. Transient expression of the IgE-BF clone in either cell type yielded IgE-BF, which potentiated an in vitro IgE response and had an affinity for lentil lectin. In contrast, when the transient expression experiments were carried out in the presence of tunicamycin, the factors no longer bound to lentil lectin. Moreover, IgE-BF expressed under these conditions suppressed an in vitro IgE response. IgE-BF lacking affinity for lentil lectin and suppressing the IgE response also resulted from transient expression of the IgE-BF gene in the presence of glycosylation inhibiting factor, a phospholipase inhibitory protein. Thus, IgE-BF that either potentiate or suppress the IgE response can be expressed from a single cloned gene; the difference in biological activities appears to be determined principally by the type of glycosylation of the common polypeptide chain. Previous work showed that IgE-BF bears an antigenic determinant recognized by the anti-Ia monoclonal antibody OX3. IgE-BF produced in the presence of tunicamycin, and IgE-BF expressed from a mutant cDNA lacking one of two carbohydrate-attachment sites, lacked the OX3 determinant. Thus, the OX3 determinant on IgE-BF appears to be associated with a site of N-linked glycosylation. PMID- 3027708 TI - Expression of the hepatitis B virus genome in chronic hepatitis B carriers and patients with hepatocellular carcinoma. AB - We examined the methylation status of CCGG sites in hepatitis B virus (HBV) DNA to determine whether methylation could be responsible for the selective expression of the HBV surface gene in chronic hepatitis B infection and hepatocellular carcinoma. Infected liver tissue from patients with low levels of viral replication was analyzed for HBV DNA copy number per haploid cell genome. Total cellular DNA, with sufficient HBV DNA, was digested with the restriction endonucleases Msp I and Hpa II, to determine whether the HBV DNA was methylated, or HindIII, to determine whether the HBV DNA was integrated or episomal. The cleavage fragments were analyzed by Southern blotting and hybridization to 32P labeled HBV DNA. In replicative chronic hepatitis B, hypomethylation of the HBV genome correlated with HBV expression in both virions and infected tissue. In carriers with nonreplicative infection, it was difficult to ascertain the role of methylation as copy number was low. HBV DNA copy number was also low in 17 out of 29 of the tumor tissues tested and as many as 14 out of 16 of the adjacent non neoplastic tissues tested. Integrated sequences were hypermethylated in the PLC/PRF/5 cell line and in six of the tumor tissues suggesting that methylation plays a role in HBV gene repression. However, since DNA from five other tumors was hypomethylated, the belief that methylation per se is an absolute determinant of HBV core gene repression does not hold for human hepatocellular carcinoma tissue. Additional factors, such as gene rearrangements, therefore, must influence HBV expression in hepatocellular carcinoma. PMID- 3027709 TI - Association between the expression of the c-myc oncogene mRNA and the expression of the receptor protein for 1,25-dihydroxyvitamin D3. AB - Studies in lymphocytes have indicated similarities in the state of activation, the time kinetics, and the pathologic states associated with the expression of the c-myc oncogene, and the expression of the 1,25-dihydroxyvitamin D3 [1,25 (OH)2D3] receptor protein. Here, we have sought evidence for an association between c-myc and the 1,25-(OH)2D3 receptor protein in mammalian cells other than lymphocytes. Comparing two rat osteogenic sarcoma cell lines, one that produces constitutively relatively high levels of the 1,25-(OH)2D3 receptor protein (ROS 17/2.8) and one in which the 1,25-(OH)2D3 receptor protein is practically undetectable (ROS 2/3), we found that the 1,25-(OH)2D3 receptor-expressing cell line also expressed c-myc mRNA. In contrast, the cell line in which the 1,25 (OH)2D3 receptor was undetectable did not express c-myc mRNA. Furthermore, we transfected mouse skin fibroblasts (NIH 3T3) with a recombinant plasmid carrying the human c-myc oncogene. We found a dramatic increase in the 1,25-(OH)2D3 receptor concentration in five separate clonal lines of NIH 3T3 cells transfected with the c-myc-carrying plasmid compared to their nontransfected counterparts or to NIH 3T3 fibroblasts transfected with the vector plasmid alone. The receptor protein of the transfected cells exhibited biochemical characteristics indistinguishable from those of classical receptors for 1,25-(OH)2D3. The increased expression in the transfected cells appeared specific for the receptor for 1,25-(OH)2D3; receptors for sex steroids were not detected in the nontransfected NIH 3T3 cells and remained undetectable after transfection with c myc. Moreover, the level of the glucocorticoid receptor protein, which was expressed in the nontransfected cells, did not change upon transfection with c myc. PMID- 3027712 TI - The regulation of collagen type IV(alpha 1) and other genes during the retinoic acid induced differentiation of wild type and mutant mouse teratocarcinoma stem cells. PMID- 3027711 TI - On the origin of induced pancreatic islet tumors: a radioautographic study. AB - Endocrine tumors of the pancreas are induced in a high percentage of young rats by injections of streptozotocin and nicotinamide (SZ/NA). Benign tumors first appear 20 to 36 weeks after drug injections. To determine the possible site of their origin, the incorporation of [3H]thymidine into islets, ducts, acini, microtumors, and gross tumors was examined by radioautography of histologic sections at 1 to 36 weeks after drug injection. Drug treatment led to early (1- to 6-week) increases in nuclear 3H labeling of exocrine pancreatic structures (ductal and acinar cells), which may involve DNA repair processes. A secondary increase in labeling of duct cells during the period of tumor emergence supports the assumption that SZ/NA-induced tumors are of ductal origin. Microtumors and gross tumors also exhibited markedly elevated rates of [3H]thymidine incorporation compared to control islets. Nontumorous islet tissue, which exhibited a gradual decrease in volume due to B-cell destruction by the drug injection, showed about 10-fold higher 3H labeling than islets of controls at all time points. The results suggest that in addition to ductal precursors, islets that survive SZ/NA-induced injury may also provide sites of focal endocrine cell differentiation to tumor tissue. Once established, both microtumors and gross tumors continue to grow by accelerated cell division. PMID- 3027713 TI - Retinoid-binding proteins and retinoid-induced differentiation of embryonal carcinoma cells. PMID- 3027710 TI - Imaging of a glioma using peripheral benzodiazepine receptor ligands. AB - Two types of benzodiazepine receptors have been demonstrated in mammalian tissues, one which is localized on neuronal elements in brain and the other, on glial cells and in peripheral tissues such as kidney. In vivo administration of 3H-labeled PK 11195 [1-(2-chlorophenyl-N-methyl-N-(1-methylpropyl)-3-isoquinoline carboxamide] or [3H]flunitrazepam with 5 mg of clonazepam per kg to rats with intracranial C6 gliomas resulted in high levels of tritiated-drug binding to the tumor as shown by quantitative autoradiography. Pharmacological studies indicated that the bound drugs labeled the peripheral benzodiazepine binding site. Binding to the peripheral benzodiazepine site was confirmed primarily to malignant cells with little binding to adjacent normal brain tissue or to necrotic tissue. Tumor cell binding was completely inhibited by preadministration of the peripheral benzodiazepine blocking agent PK 11195 at 5 mg/kg. The centrally selective benzodiazepine ligand clonazepam had no effect on PK 11195 binding to the tumor cells. When binding to other tumor cell lines grown in nude mice and nude athymic rats was evaluated, little or no peripheral benzodiazepine binding was detected on human pheochromocytoma (RN1) and neuroblastoma (SK-N-MC, SK-N-SH) tumor cells, respectively. However, high densities of peripheral benzodiazepine binding sites were observed on tumors derived from a human glioma cell line (ATCC HTB 14, U-87 MG). The presence of high concentrations of specific peripheral benzodiazepine receptors on glial tumors suggests that human primary central nervous system tumors could be imaged and diagnosed using peripheral benzodiazepine ligands labeled with positron- or gamma-emitting isotopes. PMID- 3027714 TI - Differentiated bovine parathyroid cells cultured for more than 140 population doublings show both parathyroid hormone synthesis and growth regulation by calcium ion concentration. PMID- 3027715 TI - New aspects of early embryonic induction in amphibians. PMID- 3027716 TI - Impairment of glycoprotein fucosylation in rat hippocampus and the consequences on memory formation. AB - The intraventricular injection of 2-deoxy-D-galactose led to a dose- and time dependent decrease in the fucosylation of hippocampal glycoproteins in rats whereas the incorporation of 3H-N-acetyl-glucosamine was not influenced. This effect is not related to an interference with fucose activating or transferring enzymes but can be abolished by an application of D-galactose. Thus, it seems likely that also in brain tissue a deoxy-galactose induced decrease in the fucosylation is due to a hindering of a glycosidic linkage of fucose to the deoxy sugar incorporated into glycoprotein chains. As a consequence of an intrahippocampal injection of the deoxy-sugar the retention performance of the animals in a foot-shock motivated brightness discrimination task was considerably impaired. But deoxy-galactose is effective only when administered before and immediately after training whereas either a pre- or a post-training injection did not influence the retention performance of the rats. Thus, an effective metabolic inhibition of the glycoprotein completion by the deoxy-sugar starting at the time of training seems to be crucial to interfere with such morphofunctional alterations in the neuronal network underlying the formation of a memory trace. PMID- 3027717 TI - The effects of benzodiazepines in newborn rats suggest a function for type 2 receptors. AB - In day 4 female rats lorazepam, diazepam and clonazepam produced dose-related increases in forward walking and loss of righting and diazepam produced a dose related increase in paddling. Lorazepam and diazepam increased jerks of the fore- and hind-limbs and the whole body, and clonazepam increased the latter two; these increases were not dose-related. Some doses of lorazepam and the lowest dose of diazepam increased tonic-clonic movements. Thus the benzodiazepines were observed to have two kinds of stimulant effect in day 4 rats. One is to cause hyperactivity and this effect is dose-related. The other is to cause a type of seizure-like behavior, although this action is not dose-related and the responses can be distinguished from those caused by convulsant compounds. The effects of the benzodiazepine antagonist Ro 15-1788 resembled those of the benzodiazepines. It increased hind-limb and whole body jerks, forward walking, paddling, loss of righting and tonic-clonic movements. CL 218,872, which is selective for type 1 benzodiazepine receptors, was devoid of significant effects. This suggests that the behavioral changes observed with the other compounds were mediated by the type 2 receptors. PMID- 3027718 TI - Lithium effects on adjunctive alcohol consumption. I: Comparison with adjunctive water consumption. AB - Chronic administration of lithium chloride (20 mEq/l, in drinking water) produced an earlier onset of the adjunctive consumption of both water and alcohol. Terminal consumption levels, however, were unaffected by either lithium or the liquid available for consumption. The rate of increase, once drinking was initiated, was slower for lithium subjects than it was for controls. Under extinction conditions, adjunctive alcohol consumption showed no evidence of decline for either lithium or control subjects. Water consumption by control subjects did decrease considerably as a result of extinction. Subjects receiving lithium, however, maintained their intake of water at terminal adjunctive drinking levels. PMID- 3027719 TI - Lithium effects on adjunctive alcohol consumption. II: Effects of adding concurrent shock. AB - Rats receiving chronic administration of lithium chloride (20 mEq/l) in their drinking water were tested for adjunctive alcohol (10% v/v) consumption in which temporally scheduled, noncontingent shock delivery was added following the establishment of food delivery-based adjunctive alcohol intake. The addition of shock to the eliciting schedule produced an initial reduction in alcohol consumption (Lithium subjects took longer to reach maximal suppression of drinking than did Controls), with a subsequent return to preshock levels for both groups. The reduction in alcohol consumption seen in control subjects following the discontinuation of food and shock delivery (extinction) was interpreted as suggesting that adding conflict/stress to established drinking conditions may facilitate subsequent extinction of that drinking behavior. Lithium subjects produced an initial suppression of drinking, with alcohol consumption returning to adjunctive levels by the end of the extinction series, suggesting that lithium decreases conflict/stress effects, that it impairs extinction processes, that it increases the reinforcing value of alcohol, or that it produces a combination of the three outcomes. PMID- 3027720 TI - Medial hypothalamic serotonin: effects on deprivation and norepinephrine-induced eating. AB - Evidence to date suggests an inhibitory role for serotonin (5-HT) in the regulation of feeding behavior. In the present study, hypothalamic 5-HT was investigated for its anorexic potency under different feeding conditions. In fasted rats, 5-HT (1.1-4.4 micrograms) injected into the hypothalamic paraventricular nucleus (PVN), produced a reliable dose-dependent reduction in food consumption. Under satiated conditions, this inhibitory effect was significantly larger and apparent at lower doses (down to 0.1 micrograms), in animals induced to eat by PVN injection of norepinephrine (NE). Tests with receptor antagonists, injected into the PVN immediately prior to 5-HT, revealed a dose-dependent blockade of 5-HT's action by the serotonergic blockers, metergoline, methysergide and cinanserin. While the effect of 5-HT was somewhat attenuated by administration of certain beta-adrenergic and phenothiazine-type dopamine receptor antagonists, 5-HT was totally resistant to the actions of more selective dopamine blockers and of the cholinergic and histaminergic antagonists, atropine and dexbrompheniramine. PVN injection of various serotonergic compounds, known to inhibit feeding when peripherally administered, also suppressed NE induced feeding in a dose-related manner (fluoxetine = dl-norfenfluramine greater than quipazine greater than chlorimipramine greater than dl-fenfluramine). Further tests with PVN administration of the dextro isomer and metabolite of fenfluramine showed a considerably stronger inhibitory effect with d norfenfluramine as compared to dexfenfluramine, and a particular effectiveness of peripherally injected dexfenfluramine in NE-injected rats, at doses at least 10 fold higher than centrally effective doses.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3027721 TI - Studies on a protein kinase inhibitor-insensitive, phospholipase C-sensitive pathway of lipolysis in rat adipocytes. AB - Endogenous lipid droplets were prepared by subjecting fat cells to hypotonic shock and Triton X-100 treatment. The endogenous lipid droplets were found to show lipolysis in response to epinephrine, but not to show lipogenesis from glucose in response to insulin. These results indicated that the preparation of endogenous lipid droplets did not contain any intact fat cells capable of insulin stimulated lipogenesis. Results with these endogenous lipid droplets showed that protein kinase inhibitor inhibited protein kinase-mediated hormone-sensitive lipase activity but did not reduce epinephrine-induced lipolysis. Cyclic AMP and dibutyryl cyclic AMP induced lipolytic activity in the presence of 80 mM KCl and their activities were not inhibited by protein kinase inhibitor. Phospholipase C inhibited epinephrine, cyclic AMP and dibutyryl cyclic AMP-induced lipolysis, but did not affect the lipolytic activity of either the activated or non-activated form of hormone-sensitive lipase. These results indicate the existence of a protein kinase inhibitor-insensitive and phospholipase C-sensitive lipolytic pathway in rat adipocytes. PMID- 3027722 TI - Evidence of sympathetic augmentation mediated increase in the blood and the plasma volume following vagotomy. AB - Cervical vagotomy was performed bilaterally in three series, i.e. normal (untreated), atropine treated and propranolol treated animals. The classical effect of the nerve section on heart rate, arterial blood pressure and respiration respectively were verified in the three series to confirm the reproducibility of the findings. The pre- and post-vagotomy plasma volume and haematocrit were estimated and thus the blood volume was calculated in each animal. The investigation indicated that: The interruption of vagal sensory information reflexively expanded the plasma and blood volume. This information was normally tonically active and inhibitory in nature. The volume expansion after vagotomy was brought about by stimulation of autonomic nerves, most probably the sympathetic nerves. These three conclusions suggested that expansion of vascular capacity as well as blood and plasma volume was controlled through vagal sensory information. PMID- 3027723 TI - Ontogenetic studies on mu, delta and kappa opioid receptors in rat brain. AB - The ontogenetic pattern of multiple opioid binding sites in rat brain from birth until weaning has been investigated. [3H]-dihydromorphine ([3H]-DHM)3 [3H]-D-Ala2 D-Leu5-enkephalin ([3H]-DADLE) and [3H]-dynorphin A (1-8) ([3H]-DYN) as markers of mu (mu), delta (delta) and Kappa (kappa) sites were utilized respectively. The analysis of the kinetic parameters of [3H]-DHM binding shows that, at birth, mu sites possess an affinity similar to that of adult animals, and a density of 50%, which reaches 80% of the adult value at day 4. On the contrary, [3H]-DADLE binding in the first post-natal days shows low affinity and low density and delta sites do not reach values comparable to the adult ones until the second week of life. The kinetic parameters of [3H]-DYN binding are almost undetectable during the preweanling period, due to the very low density of kappa sites at this stage of life. Displacement studies with mu-, delta- and kappa-selective ligands show that the Ki values on [3H]-DHM binding sites were similar in 4 day old and adult animals for all the tested compounds, whereas Ki values on [3H]-DADLE and [3H] DYN binding sites reflected an immaturity of delta and kappa receptors. In conclusion, our data suggest that multiple opioid receptors follow different ontogenetic patterns. In the first stages of life only mu receptors are almost mature and possibly mediate endogenous opioid actions and exogenous opiate pharmaco-toxicological effects. PMID- 3027724 TI - Broad host range vectors derived from an RSF1010::Tn1 plasmid. AB - Plasmid vector derivatives of the IncQ/P4 plasmid RSF1010 available for cloning DNA into a broad range of bacterial species were constructed. The plasmid pAYC31 constructed for the positive selection of inserted fragments contains part of transposon Tn1 inserted into the sequence of the gene sul. Gene aph transcription in pAYC31 can be initiated from the promoter for the transposase gene tnpA which is under the negative control of the gene tnpR product (Heffron, 1983). The insertion of a BamHI fragment, or of fragments generated by Sau3A, BclI, BglII, or XhoII digestion into the unique BamHI site within the gene tnpR sequence, leads to initiation of transcription from the promoter of the tnpA gene toward the aph gene. Expression of the aph gene upon insertion of a BamHI restriction fragment provides a positive selection for hybrid plasmids by plating the transformed bacteria on media with streptomycin. Versatile cloning vectors pAYC32 of 9.7 kb in length and pAYC39 of 11.3 kb in length were also constructed. Insertion into the BamHI site of vector pAYC32 of a 1.6-kb BglII fragment that contains the lambda cos site produced cosmid vectors pAYC51 and pAYC52. The two 11.3-kb cosmids differ only by the orientation of the 1.6-kb BglII fragment. By insertion into the BamHI site of pAYC32 of a BglII-BamHI fragment of plasmid pHC79 that contains the gene tet and lambda cos site cosmid vector pAYC53 was constructed. Vector pAYC31 was used to construct a gene bank from the chromosomal DNA of an obligate methylotrophic strain Methylomicrobium flagellatum KT. PMID- 3027725 TI - Plasmid pU29, a vehicle for mutagenesis of the photosynthetic puf operon in Rhodopseudomonas capsulata. AB - Plasmid pU21, which carries the reaction center and light-harvesting genes (puf operon) of Rhodopseudomonas capsulata, has been redesigned by site-specific mutagenesis. Five restriction sites have been removed and three unique restriction sites have been introduced into this 11,589-bp pBR322 derivative. The modifications divide the puf structural genes into four regions separated by five unique and nonmutagenic restriction sites. These four fragments have been subcloned into the M13-mp series of vectors to facilitate oligonucleotide mediated site-specific mutagenesis experiments on the photosynthetic apparatus structural genes. The inserts can then be returned from the M13 replicative form to the redesigned pU21 derivative. The modified plasmid, pU29, greatly facilitates in vitro mutagenesis experiments since previously described techniques and screening procedures are more efficient with M13 derivatives carrying smaller inserts. Additionally, tandem homologous sequences (the reaction center L and M subunits) within the puf operon are now separated on different phage vectors, eliminating problems encountered in the targeting of mutagenic oligonucleotides to only one of the two homologous sites. PMID- 3027727 TI - The characterization of a conjugative R-plasmid isolated from Aeromonas salmonicida. AB - We have analyzed two related R-plasmids isolated from Aeromonas salmonicida: pAr32 and pJA8102-1, which encode resistance to chloramphenicol, streptomycin, and sulfonamides (Su). The genetic map of pJA8102-1 indicated that this plasmid consists of at least three functionally different regions. One is the region conferring ability to transfer (tra), one is the region coding for drug resistance genes (r-det), and one is the region encoding the replication functions (rep). Both plasmids contain repeated sequences (RS); pJA8102-1 has four copies of this RS, while pAr-32 has three copies. The RS sequence of pJA8102 1 are located adjacent to each of the drug-resistance genes. These sequences might have been responsible for the duplication of the Su-resistance gene on this plasmid. PMID- 3027726 TI - Characterization of the Streptomyces plasmid pVE1. AB - pVE1 (11.0 kb) was isolated from Streptomyces venezuelae ATCC 14585 and characterized. pVE1 has a broad host range and an apparent copy number of about 100 during exponential growth, rising to 1500 during stationary phase. It is a pock-forming, self-transmissible fertility factor which promotes chromosome recombination in S. lividans at frequencies of about 0.1% per total parents. A detailed restriction map for 15 enzymes was determined. Genes for ThioR (thiostrepton resistance), NeoR (neomycin resistance), and pBR322 derivatives were inserted into pVE1 and the resulting plasmids were analyzed for self transmissibility and stability. The plasmid has an essential region of approximately 2.5 kb and a region of 1.0-3.6 kb required for conjugation. NeoR and ThioR vectors were constructed with unique HindIII, PvuI, BamHI, EcoRI, BglII, and EcoRV sites available for insertion of foreign DNA. PMID- 3027728 TI - Hot spot for Tn1000 insertions in cloned repressor gene of the L phage. AB - Insertions of Tn1000 into a cloned fragment of the L-phage genome comprising the repressor gene were prepared. Repressor activities produced by plasmids with insertions were assayed in vivo. As a result, the repressor gene was localized within 0.5 kb near one end of the cloned fragment. Transposon insertions were nonrandomly clustered within the repressor gene and in its close vicinity. An analysis of supercoiled plasmid DNA with the nuclease S1 revealed no distortion of the secondary structure of DNA in this region. PMID- 3027729 TI - A spontaneous insertion in the agrocin sensitivity region of the Ti-plasmid of Agrobacterium tumefaciens C58. AB - A spontaneous agrocin-resistant mutant of Agrobacterium tumefaciens strain C58 was shown to have an insertion of 1.2 kb in the agrocin-sensitivity region of pTiC58. The insertion was cloned from the Ti-plasmid, and a subclone containing only DNA internal to the insertion was used to probe the Ti-plasmid and chromosomal DNA of the wild-type strain C58. The probe showed homology to chromosomal sequences but showed no homology to wild-type pTiC58. Homology was also detected with chromosomal sequences of A. tumefaciens strains, B6, K24, and T37. These results suggest that an indigenous insertion sequence of 1.2 kb transposed from the chromosome to the agrocin-sensitivity region of the Ti plasmid in this spontaneous mutant of C58. PMID- 3027730 TI - Restriction map of virulence plasmid in Yersinia enterocolitica O:3. AB - Restriction map of the 72-kb virulence plasmid isolated from Yersinia enterocolitica O:3 was generated using EcoRI, BamHI, HindIII, and XbaI restriction enzymes. The mapping was done after cloning all of the 13 BamHI fragments of the plasmid in Escherichia coli. In addition, the restriction enzyme analysis revealed two types of virulence plasmids (types I and II) in Y. enterocolitica O:3. No functional differences between the strains bearing type I or type II plasmid were observed. PMID- 3027731 TI - The ultrastructure of normal and denervated human facial muscle. AB - The ultrastructure of normal human facial muscles from 25 nonparalytic and 17 paralytic patients revealed normal features in nondenervated human facial muscles, identical to the fine structure of other normal human and mammalian cross-striated muscle fibers. However, in denervated facial muscle, a broad spectrum of ultrastructural lesions had affected sarcomeres, abnormal inclusions, and organelles. A large variety of inclusion bodies, some of which have not been described, were also found. The spectrum of ultrastructural changes showed no dependence on the length of the denervation period. There were no inclusion bodies in all the normal facial muscle biopsies. To our knowledge, this study represents the first systematic electron microscopic investigation of normal and denervated human facial muscles. PMID- 3027732 TI - Very early correction of syndactylism. PMID- 3027733 TI - Effects of forskolin and cyclic nucleotides in animal models predictive of antidepressant activity: interactions with rolipram. AB - Forskolin, a direct activator of the catalytic subunit of adenylate cyclase (AC), and the cyclic nucleotide analogs dibutyryl cAMP (dBcAMP), 8-bromo cAMP (8 BrcAMP) and dibutyryl cGMP (dBcGMP) were tested for their ability to reverse the hypothermia or hypokinesia of mice depleted of presynaptic endogenous monoamines by pretreatment with reserpine, alpha-methyl-p-tyrosine and p chlorophenylalanine. Forskolin and the cAMP analogs decreased the rectal temperature and inhibited locomotor activity in normal mice. In mice depleted of brain monoamines forskolin reversed the hypothermia and hypokinesia; dBcAMP and 8 BrcAMP antagonized the hypothermia but were only marginally effective in reversing the hypokinesia. DBcGMP was inactive. The antihypothermic action of forskolin or salbutamol was enhanced by the novel antidepressant and cAMP selective phosphodiesterase inhibitor rolipram (4RS-[3-cyclopentyloxy-4-methoxy phenyl]-2-pyrrolidone). As an indirect effect via release of endogenous monoamines stimulating postsynaptic receptors was precluded by the monoamine depleting pretreatment, forskolin and the cAMP analogs are thought to exert their antidepressant action by directly increasing brain cAMP availability. This is achieved by forskolin via activation of the catalytic subunit of AC and by the cAMP analogs via substitution for cAMP. These findings suggest that antidepressant activity is crucially linked to enhanced cAMP availability within brain effector cells. The successful treatment of endogenously depressed patients with rolipram supports this assumption. PMID- 3027734 TI - Lithium decreases 5-HT1A and 5-HT2 receptor and alpha 2-adrenoceptor mediated function in mice. AB - Lithium administration (LiCl, 10 mmol/kg, SC on day 1, followed by 3 mmol/kg twice daily subsequently) for 14 days to mice produced attenuation of the hypothermic response to injection of 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH DPAT, 0.5 mg/kg SC). Head twitch responses to the 5-HT-receptor agonist 5-methoxy N,N-dimethyltryptamine (2.5 mg/kg IP) and to precursor loading with carbidopa (25 mg/kg, IP) and 5-hydroxytryptophan (100 mg/kg IP) were similarly attenuated. By contrast with this reduction of 5-hydroxytryptamine (5-HT) function mediated by the 5-HT1A and 5-HT2 receptor sub-types, repeated lithium administration had no effect on the motor response to a putative 5-HT1B receptor agonist 5-methoxy 3(1,2,3,6-tetrahydropyridin-4-yl)1H indole (RU 24969, 3 mg/kg IP). alpha 2 adrenoceptor function, assessed by the sedation response to clonidine (0.25 mg/kg, IP), was also attenuated by repeated lithium administration. It is proposed that these actions may explain the emergence of lithium as an adjunct to the treatment of refractory depressive illness. PMID- 3027735 TI - delta-9-tetrahydrocannabinol (THC) increases brain prostaglandins in the rat. AB - Intraperitoneal administration of 9-trans-delta-tetrahydrocannabinol (THC), the major psychoactive component of cannabis, in doses of 0.5, 1.0 and 2.0 mg/kg, produced a dose-related increase in the brain concentrations of prostaglandin (PG) E2 and PGF2 alpha in male rats 4 h after THC administration, as assessed by radioimmunoassay. A time-course investigation indicated that THC (2 mg/kg, IP) induced maximal increases in rat brain concentration of both PGs 2 and 4 h after administration; PG levels declined appreciably by 8 h and were normal by 24 h. A time-course study on the hexobarbitone (100 mg/kg, IP)-induced hypnosis potentiating effect of THC (2.0 mg/kg, IP) in male rats revealed that this pharmacological action of the cannabinoid correlated well with the time-course of the THC-induced increase in rat brain PG concentrations. The present study lends support to earlier reports contending that PGs may mediate some of the central actions of THC. PMID- 3027737 TI - [Hydroxylapatite for jaw augmentation--experience after 2 years' use]. PMID- 3027736 TI - The effect of clonidine on plasma MHPG: evidence against tonic alpha 2 adrenoceptor control of noradrenergic function. AB - The effect of the alpha 2-adrenoceptor agonist clonidine on plasma free MHPG levels was assessed in 12 normal volunteers. A significant fall in MHPG was produced by 1.5 microgram X kg-1 clonidine IV whereas saline had no effect. The peak fall in MHPG correlated strongly with the area under the curve (AUC). In addition, a strong correlation was seen between basal MHPG and the extent of the clonidine-induced fall. This suggests that plasma MHPG levels are not simply a reflection of central alpha 2-adrenoceptor function and argues against tonic alpha 2-adrenoceptor-mediated inhibitory control of noradrenergic output. PMID- 3027738 TI - Radioprotection of mouse colony forming units-spleen against heavy-charged particle damage by WR 2721. AB - The effectiveness of S-2-(3-aminopropylamino)ethylphosphorothioic acid (WR 2721) to protect against the heavy-charged particle beams with dose-averaged LET infinity's ranging from 26 to 260 keV/micron was studied using the marrow colony forming units-spleen as a model system. WR 2721 (400 mg/kg) was injected ip 30 min before whole-body irradiation in the plateau ionization region of the Bragg curve. Significant protection was observed at 26, 51, and 135 keV/micron LET values where the data were collected with 20Ne, 28Si, and 40Ar ions, respectively. The largest component of protection was the slope change, where at LET values of 26 and 51 keV/micron the DMFs (slope) were 2.1 and 2.3, respectively, which are very close to the gamma-ray value of 2.4 (gamma LET approximately equal to 0.2 keV/micron). Protection, however, decreased with increase in LET from 51 to 135 keV/micron to the DMF value of 1.2 and no significant protection was observed against 56Fe ions at 260 keV/micron. Significant increases in extrapolation number occurred with gamma rays and neon particles. The results are discussed in terms of charged particle track structure, radiation chemistry, and potential clinical applications. PMID- 3027740 TI - Alterations in alpha-adrenergic and muscarinic cholinergic receptor binding in rat brain following nonionizing radiation. AB - Microwave radiation produces hyperthermia. The mammalian thermoregulatory system defends against changes in temperature by mobilizing diverse control mechanisms. Neurotransmitters play a major role in eliciting thermoregulatory responses. The involvement of adrenergic and muscarinic cholinergic receptors was investigated in radiation-induced hyperthermia. Rats were subjected to radiation at 700 MHz frequency and 15 mW/cm2 power density and the body temperature was raised by 2.5 degrees C. Of six brain regions investigated only the hypothalamus showed significant changes in receptor states, confirming its pivotal role in thermoregulation. Adrenergic receptors, studied by [3H]clonidine binding, showed a 36% decrease in binding following radiation after a 2.5 degrees C increase in body temperature, suggesting a mechanism to facilitate norepinephrine release. Norepinephrine may be speculated to maintain thermal homeostasis by activating heat dissipation. Muscarinic cholinergic receptors, studied by [3H]quinuclidinyl benzilate binding, showed a 65% increase in binding at the onset of radiation. This may be attributed to the release of acetylcholine in the hypothalamus in response to heat cumulation. The continued elevated binding during the period of cooling after radiation was shut off may suggest the existence of an extra hypothalamic heat-loss pathway. PMID- 3027739 TI - Hydrolysis of WR2721 by mouse liver cell fractions. AB - A study of the dephosphorylation of WR2721 by broken cell preparations of mouse liver revealed the presence of at least two distinctive activities. One activity was inactivated by heat treatment and was present in the nuclear and microsomal fractions. It had an optimum pH at 9 and was inhibited by sodium vanadate, EDTA, and phenylalanine. Further subcellular fractionation demonstrated the localization of this activity in plasma membrane. A second WR2721 hydrolysis activity was detected in the cytosol fraction (postmicrosomal supernatant), which changed little with pH over the range of 5 to 10; sodium vanadate did not inhibit it. The cytosolic activity in response to heat treatment was complicated since there was an initial decrease followed by an increase in catalytic activity as a function of time at 55 degrees C. Enzyme kinetic analysis of the plasma membrane associated activity in the microsomal fraction was performed, and Km and Vmax values of 12.5 and 69.9 nmol/min/mg protein, respectively, were obtained. PMID- 3027741 TI - Free radical formation in crystals of 2'-deoxyguanosine 5'-monophosphate irradiated at 15 K: an ESR study. AB - Radiation-induced radicals in single crystals of 2'-deoxyguanosine 5' monophosphate (5'-dGMP) at 15 K have been studied by electron spin resonance (ESR) spectroscopy. At low temperatures three radicals were analyzed in detail. The negatively charged pi anion of the guanine base completely dominated the spectra. Weaker resonances were due to an alkoxy radical with the spin density in the C3'-O3' region of the sugar moiety as well as another sugar-centered radical. The anion rapidly decayed upon exposure to uv light at 15 K or by annealing above 25 K. In both cases no successor radical was observed. The second sugar-centered radical decays at 200 K with a concomitant appearance of the resonance from the C8 H-addition radical. By annealing at 295 K the latter resonance was the only one observed. After irradiation at 295 K, however, an additional resonance from a sugar-centered radical, which has been analyzed previously by B. Rakvin and J. N. Herak (Radiat. Res. 88, 240-250 (1981)) was observed. A reinvestigation of this resonance was performed. PMID- 3027742 TI - The assessment of recovery of the intestine after acute radiation injury. AB - Several aspects of intestinal function and morphology are affected by acute radiation damage, including changes in the activity of proliferative cells in the crypts, immune cell populations, and the transport of various substrates. This study was designed to compare the time course of the recovery of intestinal proliferation, transport, and leukocyte population following radiation injury. Rats received a single dose of 6 Gy to the abdomen from a 137Cs source and were studied 3, 7, and 14 days later. No changes in the passive uptake of L-glucose or D-leucine were observed in the jejunum. Active transport of D-glucose and maximal water uptake were reduced at 3 days but had returned to normal by 7 days, whereas L-leucine uptake required more than 7 days to return to control levels. Mucosal permeability, assessed by an in vivo potential difference technique, remained increased 7 days after irradiation. Ornithine decarboxylase, an indicator of DNA synthetic activity, was elevated following radiation treatment and remained so even after 14 days. By comparison, myeloperoxidase activity, used as a quantitative monitor of granulocyte numbers, was still reduced after 7 days. These data indicate that while certain parameters of gut function may return to normal soon after radiation injury, the recovery of other factors is more prolonged. Thus the return of transport function to normal values post irradiation may be viewed as an adaptive change rather than simply the recovery of the tissue. PMID- 3027744 TI - [Clinical application of the Gompertz function to small-cell bronchial cancer]. PMID- 3027743 TI - [EPR study of the blood of animals during an acute radiation lesion]. AB - The intensity of the ESR signal of methemoglobin, Fe3+-transferrin, and Cu2+ ceruloplasmin in blood and spleen of mice exposed to 6 Gy radiation was shown to undergo considerable changes at early times of acute radiation sickness. PMID- 3027746 TI - [Skeletal scintigraphy in the evaluation of the repair processes following extremity amputation with stump reconstruction]. PMID- 3027745 TI - [Roentgenographic features of fibroadenomas of various histological images in the breast]. PMID- 3027747 TI - Distribution of iodized oil within the liver after hepatic arterial injection. AB - Radiography and microscopy were used to investigate the hepatic distribution of iodized oil injected into the hepatic artery in a rabbit VX2 tumor model. Iodized oil accumulates within hepatic metastases and in a ringlike fashion around them. Radiographic and histologic appearances were correlated, and it was concluded that ringlike deposition occurs in peritumoral sinusoids. There was no evidence that iodized oil is cleared by hepatic lymphatics. Early clearance of iodized oil into bile may possibly be caused by localized hepatic ischemia from oil microemboli or by direct phagocytosis by Kupffer cells. The remaining oil is washed through hepatic vasculature, circulates systemically, and is cleared by reticuloendothelial cells in lung, spleen, liver, and bone marrow. This mode of clearance, which has not been considered previously, may be important in the prediction of toxic effects caused by lipid and lipophilic antitumor agents administered via the hepatic artery. PMID- 3027748 TI - [Axelsson's plaque formation rate index (PFRI)]. PMID- 3027750 TI - [Etched porcelain veneers]. PMID- 3027749 TI - [Material loading in ceramic restorations]. PMID- 3027751 TI - [Non-precious metals in the Konus crown technic]. PMID- 3027752 TI - [Hi-ceram]. PMID- 3027753 TI - [Addition cross-linked silicon impression materials in comparison with other elastic impression materials]. PMID- 3027754 TI - [Endosseous implants]. PMID- 3027755 TI - [Endosseous implantation procedures for non-metallic aluminum oxide ceramic implant materials]. PMID- 3027756 TI - [Metal-ceramic crowns. Preparation]. PMID- 3027757 TI - [Shaping the occlusion in cast restorations]. PMID- 3027758 TI - [The fixed torsion bridge]. PMID- 3027760 TI - [Linkage analysis and highly polymorphic DNA markers]. PMID- 3027759 TI - Comparative neurobiology of opioids in invertebrates with special attention to senescent alterations. PMID- 3027761 TI - [Growth factors of transformed cells]. PMID- 3027762 TI - [Effect of alcohol on the leukocyte system. III. Phagocytic and enzymatic activities of granulocytes in alcoholics]. PMID- 3027763 TI - Locally advanced breast cancer with inflammatory component: a clinical entity with a poor prognosis. AB - Locally advanced and inflammatory carcinomas of the breast are two distinct entities, with widely accepted differential clinical criteria. We describe a particular type of locally advanced cancer which acquires during its evolution inflammatory characteristics, although limited to a small area of skin and thus not accomplishing the common diagnostic criteria of inflammatory carcinoma. When comparing this type of tumors, named locally advanced cancer with inflammatory component, to other locally advanced cancers, some remarkable differences are found: there is a preponderance of younger patients, premenopausal or perimenopausal, with a greater percentage of poorly differentiated tumors and negative estrogen receptors. Their evolution and survival are similar to that of inflammatory carcinomas; thus, we conclude they should be considered and treated as inflammatory carcinomas. Finally, the prognostic limitations of the TNM classification are discussed and some adjustments for future classifications are suggested. PMID- 3027764 TI - Isolated intrathoracic relapse in small-cell lung cancer: incidence and implications. AB - Between 1978 and 1983, 56 patients with limited-stage small-cell lung cancer (SCLC) were treated by combination chemotherapy and moderate-dose thoracic irradiation (30 Gy). There are four survivors beyond 4 years; other patients were followed up until death, and sites of relapse documented, with five exceptions. Nine patients died with disease apparently confined to the chest. This suggests that treatments aimed at achieving better local control warrant prospective evaluation. PMID- 3027765 TI - Iodinated derivatives of the vasoactive intestinal polypeptide (VIP) are agonists at the cat pancreas and the rat submandibular salivary gland. AB - Porcine VIP was iodinated by the chloramine-T method. The reaction products, which were separated by high pressure liquid chromatography, included residual native VIP, oxidized VIP and at least two iodinated VIP species. The iodo-VIP derivatives were recognized by antibodies raised against VIP and by VIP receptors. Furthermore, they appear to be approximately equipotent agonists to VIP in activating the adenylate cyclase in membranes from the rat submandibular salivary gland and in the stimulation of pancreatic secretion in vivo. PMID- 3027766 TI - Compensatory increase in adrenomedullary angiotensin-converting enzyme activity (kininase II) after unilateral adrenalectomy. AB - Angiotensin-converting enzyme (ACE, kininase II, dipeptidyl carboxypeptidase, EC 3.4.15.1) was characterized in the adrenal medulla of male Sprague-Dawley rats. Rat adrenal medulla and lung ACE were similar in their susceptibility to Cl- activation and to the inhibition by EDTA, captopril, bacitracin and thiorphan, suggesting that rat adrenal medulla and lung ACE have similar properties. Changes in right adrenal weight and in adrenomedullary ACE activity 5 and 12 days following left unilateral adrenalectomy (UADX) were examined. Compensatory adrenocortical hypertrophy 12 days following UADX was associated with a significant increase in adrenal medullary ACE activity. This change was due not to a modified affinity of ACE for the substrate but to an alteration in ACE maximal velocity or number of available molecules. UADX had no effect on adrenocortical ACE activity. When UADX was combined with right splanchnic denervation, the increase in adrenomedullary ACE activity was blocked. The results support the existence of a functional ACE in adrenal medulla that is under neuronal control. PMID- 3027767 TI - [Biochemical utilization of the 42Ar-42K generator--measurement of specific K+ binding to Na+, K+-ATPase]. PMID- 3027769 TI - [Glioblastoma multiforme with generalized cerebral edema]. PMID- 3027768 TI - Review of analytical methods for identification and quantification of cannabis products. AB - About 100 recently published original papers, reviews, books and other communications concerning cannabis (hashish, marijuana) constituents have been reviewed with the aim of summarizing the status of analytical detection and quantitation. Detailed protocols of standard analytical methods are compared in order to recommend uniform methods for field and forensic samples and also to provide guidance to less experienced analysts in countries where cannabis sativa occurs (T. Maylon.In Big Deal: The Politics of the Illicit Drug Business, the Cannabis Commodity Market, pp. 63-107. Guernsey, London.). Because of its importance, there has been an increasing number of investigations of the chemical, botanical, pharmacological, clinical and sociological aspects of the marijuana (hashish) problem. Since analytical techniques have improved substantially during the last 10 years, many papers have been published containing a variety of methods for detection and quantification of cannabis constituents. Since the analytical situation is becoming increasingly confused and because many of the journals are unavailable in less developed countries, the aim of this paper is to give an overview of existing analytical techniques and to attempt to distinguish practical and effective methods from those which are complex or which provide questionable results. PMID- 3027770 TI - [Incidence of drug lithiasis in 1500 cases of renal calculi]. PMID- 3027771 TI - [Sonographic anatomy of the neck and its importance in lymph node staging of head and neck cancer]. AB - Our experience with sonography of the neck (about 1,500 examinations) indicates that most of the structures that are of therapeutic importance can be demonstrated by ultrasound. The cervical nerves, however, cannot be differentiated sonographically from perivascular fat because there is insufficient impedance difference presented by the myelin sheaths. The typical sonographic anatomy of the anterior and lateral cervical muscles, of the major vessels, the thyroid and parathyroid and salivary glands has been analysed. The structures that can be shown by sonography have been related to dissections of the neck. PMID- 3027772 TI - [Fascicular leiomyoma of the esophagus]. PMID- 3027773 TI - [Small bowel lymphoma as a complication of non-tropical sprue (celiac disease). A case report]. PMID- 3027774 TI - [Traumatic or inflammation-induced duodenal perforation? A case report]. PMID- 3027775 TI - [Diverticulosis of the appendix. A case report]. PMID- 3027776 TI - [Benign teratoma of the retroperitoneum in a child]. PMID- 3027777 TI - [Computed tomographic diagnosis of extra-abdominal desmoids]. PMID- 3027778 TI - [Temporal encephalocele]. PMID- 3027779 TI - [Semiquantitative SPECT of anterior dislocation of the temporomandibular joint disk]. AB - SPECT examination of the TMJ using 99m Tc-MDP was performed in 43 patients with arthrographically proven anterior dislocation of the disc and in 30 normals. The results were evaluated visually and also in a semi-quantitative manner that took account of relative 99m Tc activity in the TMJ and of the age of the patient. In the presence of arthrographically proven anterior, but reversible, disc dislocation, the semi-quantitative method proved positive in 75% of cases (28 cases). In joints with fixed anterior dislocation (29 cases), bone changes were demonstrated in 86%. Visual evaluation was positive in 50% of reversible, and in 72% of non-reversible dislocations. Semi-quantitative SPECT of the TMJ is excellent for demonstrating bone reaction resulting from TMJ dysfunction and for indicating the severity of the joint abnormality. PMID- 3027780 TI - [1st results of the diagnosis of focal liver and spleen lesions using gradient echo sequences]. AB - 15 healthy subjects and 39 patients with focal liver and spleen lesions were examined via MR tomography at 1.5 tesla. Gradient field echos at small angle excitation (less than 90 degrees) were employed. The imaging time per layer was 10 seconds so that rapid imaging could be carried out at respiratory standstill. This enabled visualisation of liver and spleen without interference by breathing artifacts and with accurate localisation. Focal lesions can be imaged best at low flip-angle pulses (liver) or low to medium-angle pulses (spleen). The primary liver cell carcinoma is visualised as an inhomogeneous structure with similar signal intensity as the surrounding tissue. All other examined liver lesions (metastases, haemangiomas, lymphatic infiltrates, echinococcus cysts, FNH, gummae) showed greater signal intensity than the remaining organ at small angle excitation. Furthermore, contrast reversals were seen at medium-angle pulses. Contrariwise, with the exception of the light-coloured spleen infarcts, spleen lesions (lymphatic infiltrate, Boeck's disease or sarcoidosis) appeared darker at all excitation angles than the surrounding tissue. PMID- 3027781 TI - [Diagnosis and therapy of non-parasitic splenic cysts in children]. AB - Splenic cysts of various types are rare surgical abnormalities. The diagnostic and therapeutic procedures are discussed with special reference to the author's clinical material consisting of six children with non-parasitic cysts. Sonography and, in special cases, CT are the most important methods of examination. The desirability of conserving the spleen, while removing the cyst, is stressed. PMID- 3027783 TI - [Accuracy of computed tomographic densitometry in the neighborhood of the pelvic bones]. AB - A phantom is described which makes it possible to correct for the change in CT values produced by soft tissues; these can be treated quantitatively. As a result of beam hardening, artefacts are produced in the soft tissues which may be hypodense or hyperdense. Rules have been established for routine densitometry in clinical use. PMID- 3027782 TI - [Results of computed tomographic diagnosis in the grading of acute pancreatitis]. AB - By means of computed tomography, it is possible to classify cases of acute pancreatitis into four grades of severity with reasonable accuracy: oedematous, sero-exudative, haemorrhagic-necrotic and abscess formation. Necrotic pancreatic tissue can be differentiated from normal tissue by means of dynamic computed tomography with a high degree of accuracy and the organ can be demarcated more clearly. Of 230 patients with acute pancreatitis, 89 were treated surgically. Correlation between the CT classification and surgical findings was achieved in 83% of the cases in the three serious grades. In 86 patients (96.6%, necrosis was correctly diagnosed. The problems that arise when trying to differentiate between the haemorrhagic-necrotic form and abscess formation are discussed, particularly when there is an absence of gas in the latter type. PMID- 3027784 TI - [Diagnosis of varices in liver cirrhosis: duplex ultrasound versus gastroscopy]. AB - This study assesses the usefulness of duplex sonography in the diagnosis of oesophageal varices. 32 patients with liver cirrhosis were studied by endoscopy and duplex sonography. There was an excellent correlation in the case of the medium-sized and large-sized varices. Only small varices could not be detected by duplex sonography. On the other hand, 3 large-sized paraoesophageal varices, which were not identified by endoscopy, were easily seen by duplex sonography. The sensitivity of duplex sonography was between 0.8 and 0.9, the accuracy between 0.93 and 0.96. PMID- 3027786 TI - [The lactating breast in the magnetic resonance tomogram. Spin-echo and fat-water images]. AB - MR criteria of a lactating mamma (15 days post partum) are imaged for the first time via spin echo technique and with fat-water images. Claear characterisation of milk in the glandular parenchyma as well as of fibrosed areals and postoperative scars was achieved. Significant additional information to the spin echo images was supplied by the fat-water images. Possible clinical uses are discussed. PMID- 3027785 TI - Non-renal urological lymphomas. AB - IVP, US and CT findings for 5 rare cases of non-renal lymphomas of the urinary tract are discussed. The 4 non-Hodgkin lymphomas (NHL) and 1 Hodgkin's disease (HD) involved the ureter (2 cases), bladder (2 cases) and renal pelvis (1 case). US and CT visualised the pyelic lesion (undetected by urography) as wall thickening and detected the two ureteral lesions (which were also revealed by antegrade pyelography for the 1 HD and by retrograde pyelography for an ureteral NHL). One bladder lesion was associated with a renal lesion (CT demonstrated retroperitoneal lymph nodes); the other was a multinodular form infiltrating the entire bladder. PMID- 3027787 TI - [Hand arteriography. Surgical indications and results]. AB - Based on experience in arteriography of the hand in 108 patients the authors present in a retrospective study the surgical indications (trauma, tumour, angiodysplasia, malformation and embolic lesion) and results of this method. The diagnostic value of arteriography of the hand is shown by means of results and examples. PMID- 3027788 TI - [Local fibrinolytic therapy of vascular occlusions in the pelvic-leg area and the upper extremity]. AB - Seventy-three patients with vascular occlusions in the pelvis or lower limbs and three patients with upper limb lesions were treated by local low dose fibrinolysin, with strict control of any possible bleeding tendencies. Adequate recanalisation was obtained in 56 patients (73.6%). In ten patients, the occlusion recurred while the patient was still in hospital. After four to six months, 37 of the 56 (67%) of the vessels were still patent. In 18 patients, peripheral emboli resulted in some deterioration, but in 15 of these cases this could be treated successfully by operation. The methods and indications of local fibrinolysis therapy and the problems associated with it are discussed. PMID- 3027789 TI - [Inadequate configurational stability of a DSA pigtail catheter as a cause of complications. Case report and study of the mechanism]. AB - There have been few descriptions extravasation into the superior vena cava and right atrium during intravenous DSA. This is seen almost exclusively when using straight catheters. The configuration of pigtail DSA catheters should reduce or prevent damage to vessel wall due to extravasation. Following an extravasation into the right atrium, which we ascribed to inadequate stability of the distal end of the pigtail catheter, we carried out in vitro experiments with seven DSA catheters from a variety of manufacturers, using an atrial model. Three of these catheters behaved in a manner that would be liable to produce extravasation. PMID- 3027790 TI - [Demonstration of brain tumors using a computer-assisted x-ray image enhancement technic]. AB - It is possible to distinguish cerebral tumours from brain tissue after the injection of contrast by using an x-ray-video chain. Weak contrast situated behind strongly absorbing bone can be demonstrated by a nontomographic method by reducing the noise level and by using a special subtraction technique designed for optimal iodine contrast. For this examination, four series of images are prepared and stored (one before the administration of contrast and three subsequently). Dynamic studies of the distribution of contrast in the intra- and extra-vascular spaces of brain and tumour are produced by subtracting the stored images. The demonstration of blood-flow dynamics improves the differentiation of the tumour and, in particular, makes it possible to distinguish the tumour from cerebral oedema. The current input into the x-ray tube is low and the skin dose on the entry side is less than 0.6R for each series. The usefulness of the method in complementing computer tomography for surgical and radiation treatment is illustrated from various types of tumour. Up to the present 35 patients have been examined by this method. PMID- 3027791 TI - [High-resolution computed tomography of peripheral facial paralysis]. AB - High resolution computer tomographic examinations of the petrous bones were performed on 19 patients with confirmed peripheral facial nerve paralysis. High resolution CT provides accurate information regarding the extent, and usually regarding the type, of pathological process; this can be accurately localised with a view to possible surgical treatment. The examination also differentiates this from idiopathic paresis, which showed no radiological changes. Destruction of the petrous bone, without facial nerve symptoms, makes early suitable treatment mandatory. PMID- 3027792 TI - [Characterization of cerebral blood flow by determining the vascular mean transit time of brain tissue using DSA]. AB - By using a recently developed algorithm it is possible to quantify the dynamic information of a DSA sequence of the brain. The theory of algorithm allows to calculate vascular mean transit from time density curves. The algorithm minimizes the problems of densitometry with regard to "quantitative DSA". There is a strong correlation between vascular mean transit times and cerebral blood flow values, and therefore the results for mean transit times also correspond to the results obtained for cerebral blood flow. By computerized postprocessing of DSA-images it is possible to generate functional images of the brain with a spatial resolution that had not been attainable so far. The images represent the distribution pattern of reverse vascular mean transit times. The results from 36 patients with proven stenoses of the cervical vessels are reported. PMID- 3027793 TI - [Functional imaging of blood flow velocity amd myocardial perfusion using the Fourier transform parameter]. AB - In the past, the radiology of the cardiovascular system has consisted of angiographic investigations which only demonstrate morphology. The introduction of digital techniques into radiology has made possible the demonstration of pure morphology and beyond that, calculations which provide functional information. We demonstrate functional images which, by using Fourier transformations, provide information on blood flow and organ perfusion. In this way, it is possible to obtain important additional diagnostic information concerning the cardiovascular system, without imposing any further stress on the patient. PMID- 3027794 TI - [Magnetic resonance tomography of the heart. Experimental study of a non-invasive characterization of myocardial tissue]. AB - In this experimental study (canine heart) a variety of pathologic conditions of the myocardium were studied using ECG-triggered magnetic resonance imaging (MRI). The purpose of this study was to examine the MR appearance and the T2 relaxation times of occlusive and reperfused myocardial infarcts, of hypertrophic cardiomyopathy, and of cardiac transplants with and without acute cardiac allograft rejection. MR images were correlated with the results of histology and tissue water content. MRI identified normal myocardium with T2 values of 38 (+/- 4.3) msec. Myocardial infarcts and acute allograft rejection had significant (p less than 0.05) longer T2 values compared to normal myocardium. On the basis of T2 relaxation times no further differentiation among different pathologic entities was possible. Therefore, it is necessary to include morphologic criteria such as myocardial wall thickness, cardiac chamber size and intracavitary blood flow signal in the interpretation of cardiac MR tomograms. PMID- 3027795 TI - Metal chelates as urographic contrast agents for magnetic resonance imaging. A comparative study. AB - Paramagnetic metal chelates including Fe-EDTA, Fe-CDTA, Fe-DTPA, and Gd-DTPA were compared as intravenous urographic contrast agents for magnetic resonance (MR) imaging in rats. Each compound, administered intravenously at three different doses (0.05, 0.1, and 0.2 mmol./kg.), was rapidly excreted through the kidneys yielding renal signal enhancement between 5 min. and 50 min. after injection. Gd DTPA was most effective vor the enhancement of the renal parenchyma and pelvis. Lesser enhancement was achieved by administration of Fe-CDTA followed by Fe-EDTA and Fe-DTPA. Enhancement of skeletal muscle and liver parenchyma could not be achieved with any of the compounds included in this study. We conclude that the iron chelates mentioned above are less effective as urographic agents for MR imaging than Gd-DTPA. PMID- 3027796 TI - [Optimization of examining sequences in MR tomography using isosignal displays]. AB - Contrast performance in MR tomography is influenced by the choice of technical parameters such as pulse repetition time (TR), echo time (TE) and inversion time (TI) in the same manner as by differences in the tissue parameters relaxation time and proton density. To optimise image contrast it is therefore suggested to preselect the imaging sequence. Such presselection is possible on the basis of isosignal visualisations S(T1, T2). In the inversion recovery sequence (IR) the isosignal images obtained with the parameters TE, TI, TR enable determination of relaxation time T1 by graphic representation, from the time of zero passage of the longitudinal tissue magnetisation. PMID- 3027797 TI - [Study of the hepatitis B virus-neutralizing response in acute infection and in patients vaccinated against the infection]. PMID- 3027798 TI - Responses of cats to nasal vaccination with a live, modified feline herpesvirus type 1. AB - Intranasal vaccination with a cold-adapted strain of feline herpesvirus type 1 (FHV-1) two days before challenge gave partial protection, and four days before challenge gave complete protection, against feline viral rhinotracheitis. Protection at this time appeared to be specific since vaccination with FHV-1 did not affect the disease caused by the unrelated feline calicivirus. The time course of onset of protection also confirmed that the protective mechanism was likely to be specific. However, six days after vaccination only low levels of FHV specific IgA and IgM antibody and of interferon were found in serum and nasal washings. In lymphocyte transformation assays neither peripheral blood lymphocytes nor tonsil lymphocytes gave a significant proliferative response in the presence of FHV antigen. Pathogenesis experiments demonstrated that the tonsil and nasal turbinates were the most important sites of virulent FHV-1 replication. Vaccination significantly reduced levels of infectious virus found in both sites. The results provide evidence that no one mechanism is responsible for protection following vaccination but local specific responses are more likely to be involved. PMID- 3027799 TI - Metabolic clearance rate of cortisol in pigs: relationship to adrenal responsiveness. AB - Experiments were conducted on 12 prepuberal (18- to 20-week-old) Landrace cross Large White gilts to establish if differences in adrenal responsiveness between individuals could be explained by differences in the metabolic clearance rate (MCR) of cortisol. Pigs with the highest (n = 6) and lowest (n = 6) cortisol concentration 60 minutes after challenge with ACTH were selected from a pool of 36 commercial pigs. Tritium-labelled cortisol was infused (17 to 27 ml h-1) continuously for 120 minutes to establish 'steady state' conditions. Blood samples (10 ml) were collected at 90, 100, 110 and 120 minutes. Replicate experiments were performed on some pigs. Classification of individual pigs as high or low adrenal responders to ACTH challenge was confirmed at the end of the clearance rate experiments. The MCR of cortisol in the group classed as low adrenal responders was 59.7 +/- 7.8 litres h-1 or 1.01 litres h-1 kg-1 (n = 7) which was not significantly different from the average MCR in the group classed as high adrenal responders 60.2 +/- 5.9 litres h-1 or 1.19 litres h-1 kg-1 (n = 10). These results suggest that the repeatable differences in adrenal responsiveness to ACTH that exist between individuals within a particular strain of pig depend on differences in the rate of synthesis of cortisol in response to ACTH stimulation, rather than on differences in its rate of metabolism. PMID- 3027800 TI - An evaluation of five serological tests for the detection of antibody to bovine herpesvirus 1 in vaccinated and experimentally infected cattle. AB - More than 300 bovine sera from a previously reported vaccination and challenge trial were tested for antibodies to bovine herpesvirus 1 (BHV1) by five serological assays: enzyme-linked immunosorbent assay (ELISA) for IgM and IgG, passive haemagglutination (PHA), and two methods of virus neutralisation (VN). In a statistical comparison of ELISA (IgG), PHA and VN results, the assays showed highly significant correlations (P less than 0.01). The sensitivities of ELISA and 24-hour neutralisation tests were similar, in contrast to passive haemagglutination and one hour neutralisation which failed to detect BHV1 antibodies in some low titre sera. PMID- 3027801 TI - Prevalence of enteroviral and parvoviral antibodies in pig sera. AB - Surveys of serum antibodies to enterovirus serotype 8 and parvovirus were conducted in pigs three to 21 weeks old using the virus neutralisation (VN), enzyme-linked immunosorbent assay (ELISA) and haemagglutination inhibition tests. Antibody levels to enterovirus were at their lowest at four to eight weeks old, increased at 14 to 16 weeks to high levels which were maintained until 21 weeks. Specific immunoglobulin G (IgG) values measured by ELISA paralleled VN values and were similar in two separate farm surveys. Measurement of immunoglobulin M (IgM) levels indicated that enterovirus infection occurred about four weeks old. Sera obtained from a large number of geographically separated farms, from abattoir bled animals and from colostrum-deprived piglets raised for 33 weeks from birth in an isolated environment were tested by VN. Results revealed that porcine enterovirus serotype 8 is spread widely throughout northern Victorian piggeries and high antibody levels prevail. In contrast to the enterovirus antibody results, parvovirus antibody levels measured in piglets declined from high levels at three to four weeks to undetectable levels from 13 weeks old. PMID- 3027802 TI - Immunofluorescence in the serological diagnosis of parainfluenza type 3 and respiratory syncytial virus infection in calves. AB - The fluorescent antibody (FA) test is compared with the haemagglutination inhibition (HI) test for parainfluenza virus type 3 (PI-3) and virus neutralisation (VN) test for respiratory syncytial (RS) virus for detection and titration of virus-specific antibodies. In experimentally inoculated calves PI-3 and RS virus FA tests detected seroconversion at the same time as HI and VN tests, however, in serially diluted sera, the FA test was positive to higher dilution. In studies with paired samples from calves from four farms with respiratory problems, the FA test gave similar results to PI-3 HI and RS virus VN tests. Large increases in antibody titre to RS virus detected by FA and VN tests indicated this was the problem on two of the farms. Individual animals showed large rises to PI-3 by FA and HI test on three farms. It is concluded that the FA test provides a rapid and sensitive alternative to the more conventional serological tests for respiratory viruses. PMID- 3027803 TI - Improved potency test for live avian encephalomyelitis vaccines. AB - Potency tests were carried out on live avian encephalomyelitis virus (AEV) vaccines. Vaccines were titrated in chick embryo brain cell cultures using an indirect fluorescent antibody test just before vaccination. Groups of chickens were inoculated orally with graded doses of each vaccine. After three weeks serum was taken from the chickens and examined for antibodies to AEV by indirect enzyme linked immunoassay (ELISA). The chickens were then challenged by intracerebral inoculation of virulent AEV and monitored for signs attributable to clinical AE. The results showed a relationship between virus content, protection and antibody development. It is recommended that serological evaluation using ELISA replace the challenge step in potency tests for live AEV vaccines. PMID- 3027804 TI - Air embolism and cardiac arrest in a patient with a pulmonary artery catheter: a possible association. AB - This patient with acute mitral insufficiency suddenly developed hemoptysis and electromechanical dissociation. A post mortem chest X-ray demonstrated a large amount of air in the right ventricular cavity. It is postulated that this air embolism resulted from a catheter-induced rupture of the pulmonary artery. PMID- 3027805 TI - The predictive and discriminative value of biologically active products of eosinophils, neutrophils and complement in bronchoalveolar lavage and blood in patients with adult respiratory distress syndrome. AB - To determine whether biologically active products of eosinophils, neutrophils and complement contribute to the development of adult respiratory distress system (ARDS) we measured eosinophil cationic protein (ECP), lactoferrin (LF) and C3a in bronchoalveolar lavage (BAL) and blood by means of radioimmunoassays. Seventeen patients served as controls. Fifteen patients were studied before and after major surgery to evaluate the influence of the surgical procedure, and 12 patients with ARDS were investigated 4-12 h after the onset of the disease. Major surgery per se significantly increased ECP in BAL, LF in serum and C3a in BAL and plasma. ECP, LF and C3a levels in BAL and blood were all significantly higher in ARDS patients as compared with levels in controls and those observed after major surgery. The higher ECP levels in BAL were associated with the more severe ARDS as was also the case for C3a in BAL and plasma and LF in serum. One out of 15 patients subjected to major surgery developed ARDS postoperatively and had very high levels of ECP, LF and C3a in BAL and blood at sampling 3 h prior to onset of ARDS, and these levels were similar to those observed in ARDS patients. One out of 12 ARDS patients died from the disease and this patient had the highest level of ECP in BAL and serum. Our results strongly support the role of activated polymorphonuclears, and notably the activated eosinophils, in the pathogenesis of ARDS. Evidence is also presented that ECP can be used as a predictor of impending ARDS. PMID- 3027806 TI - Changes in arterial and cerebral venous blood gases, cerebral blood flow and cerebral oxygen consumption at different stages of thioacetamide-induced hepatogenic encephalopathy in rat. AB - Rats were subjected to repeated intraperitoneal administrations of thioacetamide (TAA) in order to produce the following stages of hepatogenic encephalopathy (HE): the early stage, characterized by activation of brain metabolism (Group 1), the precomatose stage with impairment of brain metabolism (Group 2) and the stage of recovery (Group 3). A decrease of cerebral blood flow (CBF) of 50%, related to a drop of systemic arterial pressure and to increased hematocrit, were observed in groups 1 and 2 but not in group 3. However, the cerebral oxygen consumption (CMRO2), calculated from arterial venous difference in oxygen content and taking into account CBF, was markedly decreased in group 2, but remained unchanged in group 1. This reflects the state of cerebral metabolism at these stages. The results underscore the necessity of simultaneous monitoring of CBF and blood gases for distinguishing between the particular stages of HE revealed by biochemical tests. PMID- 3027807 TI - Rewarming in immersion hypothermia: radio-wave and inhalation therapy. AB - Anesthetized random source dogs were cooled by ice water immersion (1 degree C) to a stable core temperature of 25 degrees C, and subsequently rewarmed with warm humidified inhalation (43 degrees C, 450 cc of min ventilation/kg), radio wave induction hyperthermia (4-6 W/kg) or both therapies simultaneously. The mean time required for core rewarming to 30 degrees C was 262 +/- 29 min for humidified ventilation, 68.5 +/- 6 min for radio wave therapy (P less than 0.01), and 74.8 +/- 12 for both therapies combined (P less than 0.3 vs. radio wave). There was no tissue damage with these protocols. These data suggest radio wave heating alone is the most rapid non-invasive method for core rewarming in immersion hypothermia. PMID- 3027808 TI - Neurologic outcome following successful cardiopulmonary resuscitation in dogs. AB - Successful cardiopulmonary resuscitation necessitates that both myocardial and central nervous system function be restored with minimal long-term damage. Recent resuscitation research has emphasized minimizing neurologic damage during and after cardiopulmonary resuscitation. However, whether neurologic damage is a major cause of death or morbidity following successful cardiopulmonary resuscitation is unknown. This study examined the role of neurologic injury as a cause for morbidity and mortality following cardiopulmonary resuscitation, and if parameters used successfully during resuscitation for assessing the potential for myocardial salvage, could also be used to predict neurologic outcome. Eighty eight mongrel dogs underwent 3 min of untreated ventricular fibrillation and either 15 or 17 min of cardiopulmonary resuscitation. Twenty-four hour survivors were evaluated with a neurologic deficit scoring system. Thirty-one percent of these animals were never resuscitated. Twenty-eight percent were resuscitated, but expired prior to 24 h. Approximately half of those who expired after resuscitation died from apparent neurologic sequellae. Forty-one percent of the 88 animals survived for 24 h. Two-thirds of these survivors were completely neurologically normal, while one-third were neurologically impaired. Hemodynamic parameters useful in assessing cardiovascular prognosis were not helpful in predicting neurologic outcome. Hence, although the majority of resuscitated animals did not suffer neurologic damage, up to one-third did exhibit neurologic impairment following resuscitation. Neurologic injury is also a major contributor to early death following successful resuscitation. Hemodynamic parameters of cardiovascular recovery do not predict neurologic outcome after prolonged cardiopulmonary resuscitation. PMID- 3027809 TI - Effects of naloxone on cardiac energy metabolism in acute hemorrhage in rats. AB - It has been reported that naloxone may be useful in the treatment of hypovolemic shock. However, the effects of naloxone on cardiac energy metabolism in acute hemorrhage have not been investigated. The effects of naloxone on myocardial metabolism were evaluated in the rat during acute hemorrhage with crystalloid resuscitation. Intramyocardial high energy phosphates, pyruvate, lactate and glycogen were measured. There was no significant difference in high energy phosphates, energy charge, lactate and glycogen contents between control, 1 mg naloxone and 10 mg naloxone groups. However, pyruvate level in hearts in the 10 mg naloxone group was significantly higher than that in control group. Therefore, naloxone reduced the lactate/pyruvate (L/P) ratio. Although naloxone did not improve the mitochondrial function, it improved the oxidation-reduction state in cardiac energy metabolism. PMID- 3027810 TI - A new evaluation method for instructors of advanced cardiac life support. AB - This study evaluated the ability of a specific educational program to teach instructors of advanced cardiac life support (ACLS) how to identify errors committed by team leaders of cardiac arrest simulations. The design compared experimental and control groups for differences in identification of critical performance errors, grade assignments and errors specifically emphasized in the educational program. The group receiving the educational program documented more critical performance errors (1.70 vs. 1.10, P = 0.006), made more correct grade assignments (2.35 vs. 2.0, P = 0.026), and identified more errors that were emphasized in the educational program (3.61 vs. 2.25, P = 0.0001) than the control group. The data strongly suggests that the educational program accounted for the observed differences. PMID- 3027811 TI - The comparative effects of epinephrine versus phenylephrine on regional cerebral blood flow during cardiopulmonary resuscitation. AB - Epinephrine in larger doses than currently recommended during cardiopulmonary resuscitation (CPR) has been shown to improve cerebral blood flow (CBF) following a 10-min arrest in a swine model. The purpose of this pilot study was to measure CBF during CPR, comparing high-dose epinephrine to a pure alpha-1 agonist, phenylephrine. Ten swine each weighing greater than 15 kg, were instrumented for regional CBF measurements using tracer microspheres. CBF was measured during normal sinus rhythm (NSR). Following 10 min of ventricular fibrillation, CPR was begun and regional CBF was again measured. Following 3 min of CPR, the swine were randomized to receive either epinephrine (0.2 mg/kg), or phenylephrine (0.1 mg/kg), through a peripheral intravenous line. Regional CBF was again measured 1 min after drug administration. Regional CBF following drug administration was compared using an analysis of covariance. Adjusted CBFs are expressed in ml/min per 100 g for epinephrine and phenylephrine, respectively: left cerebral cortex (12.5 vs. 2.3, P = 0.002); right cerebral cortex (13.0 vs. 2.8, P = 0.003); cerebellum (32.9 vs. 4.1, P = 0.004); midbrain (35.7 vs. 2.6, P = 0.0004), pons (30.3 vs. 2.9, P = O.006); medulla (49.5 vs. 13.6, P = 0.02) and cervical spinal cord (49.6 vs. 14.1, P = 0.003). PMID- 3027812 TI - Effects of urinastatin on cardiac energy metabolism in acute hemorrhage in rats. AB - Urinastatin, one of the trypsin inhibitors extracted from the human urine, has been reported to be useful in treating hypovolemic shock. We studied the effects of urinastatin on cardiac energy metabolism in acute hemorrhage in rats. The hemorrhage was produced by withdrawing blood (60 ml) from the abdominal aorta, and an equivalent volume of saline and 25,000 units/kg of urinastatin were injected over 5 min. The heart was then frozen quickly with liquid nitrogen, and high energy phosphates, lactate, pyruvate and glycogen contents were measured enzymatically. There was no significant difference in either ATP, lactate or pyruvate contents of the heart between the urinastatin-treated and the control groups. However, energy charge ([ATP + 0.5 X ADP]/[ATP + ADP + AMP]) and glycogen levels in the urinastatin-treated group were significantly higher than those in the control group. These findings suggest that urinastatin might produce a metabolic improvement in mitochondrial function and suppress the anaerobic glycogenolysis induced by circulatory deficiency in an acute hemorrhagic state. PMID- 3027813 TI - Ibuprofen improves survival and neurologic outcome after resuscitation from cardiac arrest. AB - Post-ischemic inflammatory changes in the central nervous system (CNS) following cardiac arrest and resuscitation are potentially responsible for ultimate survival and much of the neurologic damage, producing greater morbidity and mortality in successfully resuscitated patients. This study was undertaken to assess the non-steroidal anti-inflammatory agent, ibuprofen, in a controlled and monitored experimental model of canine cardiac arrest and resuscitation. With the investigator blinded as to the intervention, eight of 21 dogs were randomly assigned to receive ibuprofen as an i.v. bolus (10 mg/kg) and a 6-h i.v. infusion (5 mg/kg per h). The other 13 dogs received an equivalent volume of 0.9% NaCl to serve as controls. No statistically significant differences between the two groups were detected in any pre-arrest variables. All 21 dogs were successfully resuscitated. At 24 h, dogs receiving ibuprofen exhibited 100% survival, while control dogs exhibited only 54% survival (P = 0.03). The majority of deaths for the control group occurred within the first 6 h. Neurologic deficit scores were assigned at 1, 2, 6 and 24 h after resuscitation. A general trend occurred such that dogs treated with ibuprofen improved over time, while the control dogs remained severely impaired. A significant difference in neurologic deficit score was detected at 6 h (P = 0.01). At 24 h the ibuprofen group exhibited minimal neurologic deficit (5.9 +/- 3.2), and the control group exhibited significantly more severe neurologic impairment (52.2 +/- 13.0, P = 0.01). These results suggest that ibuprofen may be helpful in the pharmacologic management of cardiac arrest as a means of increasing survival and decreasing neurologic impairment. PMID- 3027814 TI - Biochemical markers in a porcine model of adult respiratory distress syndrome induced by endotoxemia. AB - To evaluate the influence of a continuous endotoxin infusion on different hematological and biochemical variables which might underlie endotoxin-induced ARDS we investigated 2 groups of pigs, one with and one without endotoxin. Complement activation - both via the classical and alternative pathways -- antithrombin III, (AI-III) factor VIII-related antigen and fibronectin levels could not precisely predict the development of ARDS in the individual animal. It was found, however, that the animals which reacted with a severe endotoxin induced pulmonary dysfunction (pulmonary responders) had significantly higher levels of complement of both the classical and alternative pathways, greater concentrations of AT-III and factor VIII-related antigen and higher fibronectin levels. These findings might probably be explained by a greater reactivity, i.e. synthesis and release, of these biochemical components in the animals which responded with a severe pulmonary reaction, a phenomenon which probably concealed the consumption. A greater "reactivity" regarding the drop in polymorphonuclear cell count was found in pulmonary responders; a phenomenon which, however, was not compensated for by increased production. A finding of presumably great clinical importance was that animals surviving the observation period had significantly higher levels of fibronectin already at baseline and throughout the observation period. PMID- 3027815 TI - Prediction of subclavian vein location using plain chest radiography. AB - The relationship between the right subclavian vein and the thoracic inlet below the clavicle was studied by Venography in 72 patients. The area of the thoracic inlet below the clavicle was defined as a radiolucent area surrounded superiorly by the lower border of the clavicle, inferiorly by the inner margin of the first rib and medially by the lateral margin of the manubrium (CRM area). In 10 patients, the subclavian vein was situated below the axis of the clavicle, and the CRM area was large enough to extend near the top of the first rib arch. In 62 patients, the subclavian vein extended above the axis of the clavicle and the CRM area was small or invisible. The existence of a large thoracic inlet below the clavicle (large CRM area which extends near the top of the first rib arch) may be a useful indicator for predicting the low location of the subclavian vein, and may be used to predict or explain venipuncture failure using the standard infraclavicular approach. PMID- 3027816 TI - Ultimate survival from septic shock. AB - Detailed hemodynamic course during prolonged (less than 12 h) septic shock was studied in 23 patients, of whom 12 ultimately died from sepsis. Hemodynamic presentation was similar in fatalities and in survivors, except for a higher heart rate and a markedly higher blood lactate in fatalities (8.4 +/- 1.4 vs. 3.7 +/- 0.4 mEq/l, P less than 0.01). During fluid resuscitation, the pulmonary artery occlusive pressure associated with the highest left ventricular stroke work was usually around 17 mmHg, but in some patients above 20 mmHg. During the course of septic shock, left ventricular function improved in both fatalities and survivors, so that an altered cardiac function had no ultimate pejorative implication. The higher blood lactate in the absence of a different hemodynamic pattern tends to indicate that peripheral distributive defect remains the essential anomaly in septic shock. Arterial lactate appears to represent the most reliable indicator of ultimate prognosis in septic shock. PMID- 3027817 TI - Hyperkalemic cardiac arrest during an infusion of potassium chloride following an overdose of propranolol. PMID- 3027818 TI - [Increased cardiovascular responses to norepinephrine in hypertrophic cardiomyopathy]. PMID- 3027819 TI - [Angiotension converting enzyme inhibitors in arterial hypertension]. PMID- 3027820 TI - [Current trends in dietetics in diabetology and their experimental bases]. AB - The last decade has witnessed drastic changes in our views on diet for diabetics, whether insulin-dependent or not. To bring blood glucose levels down to normal values, thereby preventing diabetic microangiopathy or alleviating its course, remains the compelling purpose of treatment, but the modalities and constraints of the dietetic measures which contribute to this result have been radically revised. Leaving aside fashions and controversies, three tendencies have emerged: the low carbohydrate diet does not improve the glycaemic balance but implies an excessive fat intake which may aggravate the microangiopathy. Of course, an hypocaloric diet remains fundamental in the management of obese non-insulin dependent diabetics; the effects on glycaemia of the carbohydrate ration constituents must be reconsidered. The classical distinction between "fast" and "slow" sugars seems to be excessive and insufficient, if not erroneous. Food replacements must take into account the glycaemic index; a minimal dietary fibre intake has a favourable effect on post-prandial glycaemia and lipid metabolism and is to be recommended, notably to diabetics. PMID- 3027821 TI - [Pharmacokinetics of enalapril]. AB - Enalaprilate, an antihypertensive agent that inhibits angiotensin-converting enzyme activity, is not directly absorbable. It is therefore administered as an inactive precursor, an enalaprilate ester which is hydrolysed in vivo with an ultimate bioavailability of 30-40 p. 100. Enalaprilate is entirely excreted through the kidney. With repeated administrations steady state is rapidly reached, and the drug does not accumulate. The effective half-life is 10 hours. Kinetics are linear and depend on renal function. These data, obtained in healthy volunteers with 10 mg doses, also apply to hypertensive patients receiving 20 mg. Following a 20 mg per day dose a more than 50 p. 100 inhibition of the angiotensin-converting enzyme is maintained for 24 hours. In patients with moderate renal impairment or in elderly people (creatinine clearance greater than 30 ml/min), plasma concentrations are slightly increased and there is no need for dosage adjustment. In patients with severe renal impairment (creatinine clearance less than or equal to 30 ml/min) plasma concentrations are considerably increased with a risk of strong accumulation, and dosage must be reduced to 5 or 2.5 mg per day. PMID- 3027822 TI - [Regional blood flow in congestive cardiac failure and inhibition of angiotensin converting enzyme]. AB - In congestive cardiac failure myocardial deficiency is accompanied by neurohormonal dysfunction with reflex stimulation of the sympathetic nervous system and the renin-angiotensin-aldosterone system (RAAS). These two systems act synergistically, resulting in peripheral vasoconstriction with increased vascular resistance and therefore greater demand on left ventricular haemodynamics. The role of the RAAS is better understood when angiotensin-converting enzyme inhibitors (ACEI) are used. The vasodilatation produced by these inhibitors mostly affects those vascular beds that depend on the vasopressor effect of angiotensin II, and primarily the renal vascular system: glomerular filtration is improved, and sodium excretion is increased. The RAAS has little influence on the other regional blood flows, so that the administration of ACEI is not followed by cutaneous, muscular or visceral vasodilatation. However, the cerebral blood flow remains normal or even increases despite the fall in systemic arterial pressure, and the coronary output is preserved. Angiotensin-converting enzyme inhibitors produce a strong, though selective, vasodilatation without reflex tachycardia, benefiting primarily the kidneys. The resulting increase in sodium excretion contributes to the long-term effect of ACEI in the treatment of congestive cardiac failure. PMID- 3027823 TI - [Herpes genitalis, in prostitutes. Detection of asymptomatic viral excretors and their epidemiological role]. PMID- 3027825 TI - Liver resection in malignant disease. AB - As more surgeons become familiar with the techniques of hepatic resection and the mortality and morbidity decrease, the indications for resection of malignant disease within the liver broadens. The preoperative assessment of malignant liver lesions, as well as the definition of resectability, are outlined. Indications for operative intervention as well as the results obtained are covered. The personal experience of the authors at the Royal Postgraduate Medical School Hepatobiliary Unit, Hammersmith Hospital, in dealing with malignant lesions of the liver is detailed with respect to procedures performed and postoperative morbidity and mortality. Hepatocellular carcinoma, hilar cholangiocarcinoma, and metastatic colon carcinoma are discussed in detail. The authors' experience with each of these diseases is presented. PMID- 3027824 TI - [The ten leading work-related diseases and injuries proposed by the National Institute for Occupational Safety and Health (NIOSH)]. AB - Ten leading work-related diseases and injuries were proposed by the NIOSH to be controlled. These diseases and injuries are occupational lung diseases, musculoskeletal injuries, occupational cancers (other than lung), amputations, fractures, eye loss, lacerations, and traumatic deaths, cardiovascular diseases, disorders of reproduction, neurotoxic disorders, noise induced loss of hearing, dermatologic conditions, and psychologic disorders. Current topics regarding these diseases and injuries are discussed. PMID- 3027826 TI - Intraoperative radiotherapy in stage I and II lung cancer. AB - Lung cancer is the leading cause of cancer deaths in both men and women in the United States. Treatment depends on the type and stage of lung cancer. For stage I and II cancer, surgery is usually the treatment of choice. Radiation therapy is used in patients who are considered poor risks for surgical resection. Intraoperative brachytherapy is an effective alternative to external irradiation in this group of patients. From 1958 to 1984, 55 patients with non-small-cell lung cancer were explored at Memorial Sloan Kettering Cancer Center and found to have surgical stage I or II tumors, which were considered to be unresectable mainly because of severe obstructive pulmonary disease precluding adequate resection. All these patients were treated with intraoperative brachytherapy at the time of the thoracotomy. Forty-four percent of these patients received in addition external irradiation, mainly to the mediastinum. The overall 5-year survival calculated by the Kaplan-Meier Method was 32%, and the local disease free survival was 63%. Cox regression multivariant analysis demonstrated that there is a distinct subgroup with a better prognosis based on tumor site and patient's age--ie, patients who were younger than 58 years of age and had right side lesions. PMID- 3027828 TI - Current role of chemotherapy in the management of ovarian cancer. AB - A discussion of the current role of chemotherapy in the management of ovarian cancer, based on results obtained from studies by Kurume University Hospital and the Japan Society of Obstetrics and Gynecology (JOG) group, is presented. Higher response rates were obtained for epithelial cancer when combinations of cisplatin, adriamycin, and cyclophosphamide were used. However, cancers such as ovarian cancer that form huge tumor masses cannot be cured completely with chemotherapy alone. Therefore, we think that effective combinations of chemotherapy and surgical treatment are the key to improving patients' prognoses. PMID- 3027827 TI - Chemotherapy for gestational trophoblastic tumors. AB - Virtually 100% of patients with nonmetastatic and low-risk metastatic trophoblastic tumors are now curable by only conventional chemotherapeutic agents such as methotrexate, actinomycin D, or their combination with or without other agents. However, approximately 30% of high-risk patients with metastatic trophoblastic tumors are resistant to conventional chemotherapy. All of the high risk patients resistant to chemotherapy are those with metastatic choriocarcinoma. Thus, the efficacy of multidrug chemotherapy and combination chemotherapy of cisplatin or VP16-213 with other agents is now being examined. PMID- 3027829 TI - [The prescription of analgesics in current dental practice]. PMID- 3027830 TI - First International Symposium on Itraconazole. Oaxaca, Mexico, October 7-8, 1985. Proceedings. PMID- 3027831 TI - Itraconazole in the treatment of superficial mycoses: an open trial of 40 cases. AB - Forty patients with superficial mycoses were treated with itraconazole, a new triazole derivative, in an open study. In 29 cases the diagnosis was dermatophytosis; in five, candidosis; and in six, pityriasis versicolor. Each patient received 50 mg of itraconazole orally once a day for 30-60 days. Thirty three patients completed therapy; at the end of therapy, 21.2% were considered clinically cured and 57.6% were considered mycologically cured. Results of direct microscopic examination were negative at 30 days for the patients with pityriasis versicolor; none of the patients with candidosis responded to therapy. Pruritus and other symptoms or signs of infection improved after the 15th day, but even after 60 days of therapy, nine of the 10 patients who finished the trial at that time had residual lesions. The drug was well tolerated. It appears that a dosage of 50 mg of itraconazole daily is not adequate for the treatment of these mycoses. PMID- 3027832 TI - Itraconazole in the treatment of dermatophytoses: a comparison of two daily dosages. AB - Itraconazole, the first orally active triazole antifungal agent, was tested in 185 cases of acute or chronic (recurrent) dermatophytosis. The skin infections were divided into two major diagnostic groups: tinea corporis/tinea cruris (91 cases) and tinea pedis/tinea manuum (94 cases). Each patient was randomly assigned to a regimen of 50 or 100 mg of itraconazole daily. Of the cases of tinea corporis and/or tinea cruris, 94% responded to 50 mg of itraconazole daily and 96% to 100 mg daily. The effects of therapy were evident earlier when 100 mg daily was given. Of the cases of tinea pedis and/or tinea manuum, 80% responded to 50 mg of itraconazole daily and 94% to 100 mg. Again, improvement was seen sooner with the 100-mg regimen. Only five patients (2.9%) reported minor adverse reactions. Hematologic and blood-biochemical parameters were monitored before treatment and at biweekly intervals thereafter in 37 patients; no significant abnormalities were observed. Thus, either 50 or 100 mg of itraconazole daily constitutes effective treatment for dermatophytoses. Since the larger dosage induces a faster response, it should be evaluated further. PMID- 3027833 TI - Treatment of dermatophytoses and pityriasis versicolor with itraconazole. AB - Thirty patients (18 males and 12 females) with dermatophytoses were treated with itraconazole. Twenty-eight patients received 100 mg per day for 14 days, and two patients, one with tinea pedis and one with tinea manuum, were treated for 28 days. Thirty patients (11 males and 19 females) with pityriasis versicolor were treated with 200 mg of itraconazole. for five days. Half of the patients received the 200 mg in one dose, and the other half received 100 mg twice a day. Twenty nine of the 30 patients with dermatophytoses were clinically and mycologically cured, and one patient was improved. All 30 patients with pityriasis versicolor experienced complete clinical and mycologic cure; no difference was noted between the two groups. Ten of the 60 patients treated reported adverse reactions to therapy. The primary adverse effects were nausea and epigastric pain; one patient had diarrhea. There were no alterations in the patients' blood biochemical values. These results suggest that itraconazole is safe and effective for the treatment of dermatomycoses. PMID- 3027834 TI - Treatment of chronic dermatophytosis and chronic oral candidosis with itraconazole. AB - Itraconazole was evaluated in patients with forms of superficial fungal infection that were previously unresponsive to treatment. Two groups of patients were treated, those with chronic dermatophytosis caused by Trichophyton rubrum affecting the body (10) or palm (five) and those with chronic oral candidosis (eight). All of the patients with dermatophytosis achieved complete remission in a mean period of 9.1 weeks, with three subsequent relapses. Seven patients with oral candidosis had originally presented with chronic mucocutaneous candidosis; recurrent or persistent oral candidosis had developed after initially successful treatment with ketoconazole. All of the patients with chronic oral candidal infections responded to itraconazole after a mean treatment period of 5.4 weeks, with two subsequent relapses. These results indicate that itraconazole is effective in treating these recalcitrant superficial mycoses and should be assessed in a larger unselected group of patients and compared with alternative drugs. PMID- 3027835 TI - Itraconazole in the treatment of tinea corporis: a pilot study. AB - In this pilot study, itraconazole was administered to 18 patients with tinea corporis. The dosage was 100 mg daily for four to six weeks. Follow-up visits took place two, four, and six weeks after the first dose. Skin samples were cultured, and clinical symptoms were assessed at each visit. At two weeks clinical symptoms were significantly reduced in all patients, and cultures were negative in approximately 50%. After four and six weeks, respectively, 83% and 89% of patients had negative cultures and 89% and 94% were considered markedly improved or cured in an overall assessment. None of the patients reported any adverse reactions. No changes were observed in hematologic and biochemical values except in one patient who had asymptomatic elevations in hepatic transaminases at six weeks. Itraconazole appeared to be effective in the treatment of tinea corporis. Two weeks of treatment was effective in many patients, and it was not necessary to treat any patient for more than four weeks. PMID- 3027837 TI - Itraconazole in pityriasis versicolor. AB - Forty-two patients with pityriasis versicolor were treated with itraconazole. Patients were allocated randomly to one of two groups; the first group (22 patients) received 200 mg of itraconazole per day for five days, and the second group (20 patients) received 100 mg per day for the same period. The study lasted 30 days. Each patient's clinical history was recorded before treatment. Wood's light examination was also done at this time and was repeated weekly. Direct microscopic examination of lesions was undertaken at the beginning, middle, and end of the trial. Specimens for cultures could be obtained from only 24 patients. The cure rate was 95% in the first group and 75% in the second. The difference between these rates was not significant, but the groups were small and the difference in dosage may ultimately be meaningful in terms of rates of cure. Itraconazole appears to be effective in a high percentage of cases of pityriasis versicolor. PMID- 3027836 TI - Randomized comparative clinical trial of itraconazole and selenium sulfide shampoo for the treatment of pityriasis versicolor. AB - Forty patients with pityriasis versicolor, fluorescence of involved areas under Wood's light, and positive microscopic identification of Malassezia furfur were randomly assigned to treatment with either oral itraconazole (200 mg once daily for five consecutive days) or 2.5% selenium sulfide shampoo (once daily application for seven days). Each treatment group consisted of 20 patients. On assessment three weeks after the end of treatment, all patients given itraconazole showed a response: 17 were healed, and three had mild residual lesions. Likewise, all patients in the selenium sulfide group responded to therapy: 16 were healed, and four had mild residual lesions. Tolerability, acceptability, and compliance were excellent with itraconazole. All 20 patients given the drug stated a preference for oral treatment. In the selenium sulfide group, five patients (25%) had adverse reactions attributable to the medication; one of these patients experienced an irritation severe enough to warrant the discontinuation of treatment on the third day. Ten patients in this group stated a preference for oral treatment. PMID- 3027838 TI - Pityriasis versicolor: efficacy of two five-day regimens of itraconazole. AB - An open, comparative trial of two brief regimens of itraconazole for pityriasis versicolor was carried out. Twenty-four patients received 100 mg twice a day for five days, and 23 received 100 mg once a day for five days. Patients were enrolled in the study on the basis of the presence of clinical symptoms and positive findings on direct microscopic examination. Tolerance to the drug and mycologic and clinical outcome were assessed on days 6, 14, and 28 after the initiation of treatment. For 23 patients in each group, skin samples were negative for fungi by day 18. Clinical improvement was slow; first scaling disappeared and then erythema and inflammation. Dyschromia persisted during the observation period. Six patients had adverse reactions to treatment; none discontinued treatment. PMID- 3027839 TI - Itraconazole in pityriasis versicolor: ultrastructural changes in Malassezia furfur produced during treatment. AB - Twenty-eight patients with pityriasis versicolor were treated orally with 200 mg of itraconazole per day in an open, randomized comparison of five-day and seven day treatment regimens. The morphologic changes in Malassezia furfur produced by treatment were confirmed by studies with transmission and scanning electron microscopy. The clinical results showed that seven days of treatment were more effective than five days. Transmission electron microscopy revealed cytopathic changes in the fungus at the end of treatment, intracellular necrosis being complete within seven to 28 days. Cell-surface alterations detected by scanning electron microscopy developed more slowly--maximal changes were observed 14 days after the beginning of treatment. One patient developed nausea during treatment; no other adverse effects were attributed to the drug. PMID- 3027840 TI - Randomized comparative trial of three regimens of itraconazole for treatment of vaginal mycoses. AB - A randomized comparative study of three treatment regimens with itraconazole was carried out in 60 nonpregnant women with acute vaginal candidosis. Vaginitis was demonstrated by both a positive culture and positive findings on microscopic examination of a vaginal smear as well as by the presence of clinical symptoms. Sixty patients seen over a three-month period were randomly allocated to receive one dose of 200 mg daily for two consecutive days (regimen A), 200 mg twice a day for one day (regimen B), or 200 mg once a day for three consecutive days (regimen C). Each group comprised 20 patients. In group A, 65% were clinically and microbiologically cured, 5% were clinically but not microbiologically cured, and 30% relapsed. In group B, 55% were clinically and microbiologically cured, 10% were clinically but not microbiologically cured, 15% did not respond to treatment, and 20% relapsed. In group C, 75% were clinically and microbiologically cured, 10% did not respond, and 15% relapsed. PMID- 3027841 TI - Itraconazole in the treatment of vaginal candidosis and the effect of treatment of the sexual partner. AB - The efficacy of itraconazole in the treatment of vaginal candidosis and the effect of treatment of the sexual partner were evaluated. Women received 100 mg of itraconazole daily for five days. The sexual partners of one-half of the women were treated with the same dosage of itraconazole; the other men received placebo. Study design was double blind. Sexual relations were discontinued during treatment. On day 0 and day 30 after initiation of treatment, laboratory values were determined. At 30 days, 14% of the women whose sexual partners had received placebo and none of those whose sexual partners had received itraconazole had relapsed. These differences, however, were not statistically significant. PMID- 3027842 TI - Itraconazole in the treatment of human mycoses: review of three years of clinical experience. AB - During the first three years of clinical investigation of itraconazole, more than 1,000 patients with mycoses were treated with the drug. Almost 50% were women with vaginal candidosis; a dosage of 200 mg per day for three days appeared to be optimal for their treatment. Several treatment regimens were tested for pityriasis versicolor; the minimum total dose necessary for optimal results was 1 g. A randomized comparison of 50-mg and 100-mg daily doses for the treatment of skin mycoses indicated that the optimal dosage is 100 mg. The results of short courses of treatment for superficial dermatophytoses suggest that such regimens may be effective, and the results of an ongoing double-blind comparison of itraconazole and griseofulvin suggest that itraconazole is superior in these infections. The outcome of treatment of systemic mycoses with itraconazole, especially sporotrichosis, chromomycosis, and aspergillosis, indicates that itraconazole may be useful in therapy for life-threatening fungal infections when standard therapy has failed. PMID- 3027843 TI - Activity of orally, topically, and parenterally administered itraconazole in the treatment of superficial and deep mycoses: animal models. AB - The activity of itraconazole in vitro was evaluated for 2,094 strains of 132 fungal species, one achloric alga, nine actinomycetes, and six bacterial species. Itraconazole was active against dermatophytes (271 strains), Candida species (1,303), Cryptococcus neoformans (27), Torulopsis species (170), Pityrosporum species (40), Aspergillus species (87), Sporothrix schenckii (12), dimorphic fungi, Dematiaceae, and various other organisms. This activity depended largely on the test conditions and the medium used. Ittraconazole was as active as ketoconazole in the treatment of dermatophytoses and of both superficial and deep candidosis at oral doses about eight and four times lower, respectively, than the doses of ketoconazole required. Disseminated dermatophytosis was cured more rapidly by itraconazole than by ketoconazole. Parenteral and oral itraconazole were of equal efficacy for the treatment of systemic candidosis. Itraconazole used topically was more active than reference compounds against microsporosis, trichophytosis, and superficial candidosis. Given orally, itraconazole was effective therapy for aspergillosis and meningo-cerebral cryptococcosis in mice and for generalized cryptococcosis, histoplasmosis, and sporotrichosis in guinea pigs. No drug-related adverse effects were observed. PMID- 3027844 TI - Degenerative changes in fungi after itraconazole treatment. AB - Changes in morphogenetic behavior and structural degeneration after exposure to itraconazole are illustrated in Candida albicans, Cryptococcus neoformans, Pityrosporum ovale, Paracoccidioides brasiliensis, Trichophyton rubrum, and Aspergillus fumigatus. With the exception of P. ovale, primary alterations are seen at the cell periphery and the cytoplasmic vacuoles in which lipid-like vesicles assemble. These changes are usually accompanied by a marked increase in cell volume, impaired cell division, or abortive hyphal outgrowth. The concentration of itraconazole necessary to induce irreversible structural degeneration (necrosis) depends greatly on the species used, the time of incubation, and the morphogenetic form in which the fungus is grown and varies from 10(-10) M (P. brasiliensis) to greater than 10(-6) M (C. albicans). Itraconazole achieves these effects either at a concentration comparable to that required for ketoconazole (C. albicans and C. neoformans); at concentrations 10- to 100-fold lower (P. ovale, T. rubrum, P. brasiliensis), or at concentrations 100-fold lower (A. fumigatus). PMID- 3027845 TI - Itraconazole: pharmacologic studies in animals and humans. AB - Several pharmacologic studies of itraconazole, an orally active antifungal triazole, were conducted in humans and animals. In dogs and rats, no significant toxic effects were seen at doses of up to 40 mg/kg. Endocrinologic studies demonstrated that itraconazole, unlike ketoconazole, does not significantly affect human testicular and adrenal steroidogenesis. Pharmacokinetic studies indicated that itraconazole has a high tissue affinity and a longer half-life than ketoconazole. These pharmacologic observations suggest that itraconazole has a broader spectrum of activity than ketoconazole and a lower potential for producing adverse effects. PMID- 3027846 TI - Oral treatment of paracoccidioidomycosis and histoplasmosis with itraconazole in humans. AB - Twenty-five patients with paracoccidioidomycosis and 17 patients with histoplasmosis were treated with itraconazole. All patients were adults. Those with paracoccidioidomycosis exhibited the chronic disseminated form of the disease; 21 of these patients had lesions in two or more locations, and four had lesions only on the larynx or mouth. Itraconazole was administered at a daily dosage of 50 mg for six months in the majority of these cases. All infections were clinically cured or showed striking improvement. Patients with histoplasmosis had the chronic pulmonary or chronic disseminated form of the disease. A daily dose of 100 mg was administered until clinical cure was established; the dose was then changed to 50 mg until the completion of six months of treatment. Twelve infections were clinically cured; four were strikingly alleviated. The remaining patient, who discontinued treatment with itraconazole after two months, had a severe relapse and died of respiratory failure. PMID- 3027847 TI - Itraconazole in the treatment of paracoccidioidomycosis: a preliminary report. AB - The preliminary results of itraconazole therapy in 16 patients with active paracoccidioidomycosis were evaluated. The course of therapy--itraconazole administration for six months at a dose of 100 mg per day--was completed in 13 cases. This new triazole appeared as effective as ketoconazole in reducing the symptoms and arresting the progression of the mycosis. The scoring system employed to evaluate the effect of the drug showed that the condition of no patient worsened or remained the same during therapy. On the contrary, 11 (84.6%) of the 13 patients experienced major improvement and the other two (15.4%), minor improvement. No adverse reactions were reported by the patients, and there were no toxic effects on bone marrow or liver. Although experience with itraconazole is still limited, results to date indicate that this new drug is safe and effective for the treatment of paracoccidioidomycosis. Further trials with shorter periods of therapy seem warranted. PMID- 3027848 TI - A clinical trial of itraconazole in the treatment of deep mycoses and leishmaniasis. AB - Itraconazole was administered orally to two patients with sporotrichosis, 10 patients with paracoccidioidomycosis, three with mycetomas (due to Madurella grisea, Streptomyces madurae, and Pseudochaetosphaeronema larense, respectively), nine with chromomycosis due to Cladosporium carrionii, five with chromomycosis due to Fonsecaea pedrosoi and five with leishmaniasis (including one with the nodular disseminated form). The clinical and laboratory tests showed excellent tolerance to the drug with a total absence of adverse reactions. Satisfactory results were achieved against paracoccidioidomycosis, sporotrichosis, and chromomycosis due to C. carrionii (apparent cure was achieved in a short time). Encouraging improvement was noted in the treatment of mycetoma due to M. grisea. Among the five cases of leishmaniasis, a complete clearing was achieved in one and an encouraging improvement in two, including the one with the nodular disseminated form. Two patients with F. pedrosoi infection were apparently cured after the addition of thermotherapy and flucytosine, respectively, to the treatment regimen. PMID- 3027849 TI - Itraconazole for deep mycoses: preliminary experience in Mexico. AB - Itraconazole has been used as oral therapy for deep mycoses since July 1984 in our institution. So far, therapy has been evaluated for eight patients: one with cutaneous coccidioidomycosis; two with chromomycosis (Fonsecaea pedrosoi); and five with sporotrichosis (three fixed, one lymphangitic, and one disseminated). Clinical and mycologic evaluations were done at the beginning of treatment, 15 days after the initiation of treatment, and monthly thereafter. Dosage was 100 200 mg per day, and duration of treatment was based on response. The cases of coccidioidomycosis and lymphangitic sporotrichosis were mycologically cured after two and seven months of treatment with 100 mg per day, respectively. The six other patients required higher doses. Two patients with sporotrichosis and one with chromomycosis were cured, and one patient with sporotrichosis and one with chromomycosis were markedly improved. In one patient with sporotrichosis, treatment was discontinued because of the slow response. No adverse reactions to treatment were reported. PMID- 3027851 TI - Initial experience in therapy for progressive mycoses with itraconazole, the first clinically studied triazole. AB - The search for antifungal drugs, particularly oral agents with a broad spectrum of activity, led to the study of itraconazole, a triazole. In vitro susceptibility testing suggested activity against a variety of fungal pathogens affecting humans. Pharmacokinetic studies indicated absorption after oral administration to patients, with prolonged serum concentrations and peak levels exceeding the in vitro inhibitory concentrations for most pathogens. Twenty-four patients have been enrolled in efficacy studies; two-thirds of these individuals had experienced a relapse after apparently successful therapy with other drugs, had failed to respond to other drug therapy, and/or had experienced intolerable side effects with other drugs. Fourteen patients have thus far responded to treatment with itraconazole; this group includes two persons each with coccidioidal pneumonia and adenopathy, pulmonary coccidioidomycosis, and dermatophytosis plus onychomycosis as well as one person each with coccidioidal epididymitis, adrenal histoplasmosis, head and neck histoplasmosis, cutaneous blastomycosis, oral-sinonasal alternariosis, chromoblastomycosis, invasive pulmonary aspergillosis with fungus ball, and onychomycosis. Five other patients have had partial responses to therapy, three have failed to respond, and two have been deemed unassessable. Drug toxicity has been limited in 211.8 patient-months of therapy. The results are encouraging and indicate that more extensive studies are warranted. PMID- 3027850 TI - Early experience with itraconazole in vitro and in patients: pharmacokinetic studies and clinical results. AB - The efficacy of itraconazole, a new triazole antifungal agent, was studied in vitro and assessed in patients. The MICs of itraconazole for 16 strains of Aspergillus fumigatus were in the same range as those of amphotericin B: less than 0.09-0.36 microgram/ml vs. less than 0.09-0.78 microgram/ml, respectively. Eight adult patients with systemic fungal infections were treated orally with 100 200 mg of itraconazole two times a day. A patient with relapsing histoplasmosis (Histoplasma capsulatum var. duboisii) was cleared of the infection; a patient with arthritis of the knee due to Phialophora parasitica did not respond to treatment; four patients with semiinvasive pulmonary aspergillosis improved dramatically and were considered clinically cured; and two patients with aspergilloma improved. The duration of follow-up was one to nine months. Levels of itraconazole in body fluids were measured by a bioassay. Levels of drug in knee fluid were about 30% of the simultaneous levels in plasma. A progressive decrease in the level of itraconazole in plasma occurred in two patients, and a progressive increase in the levels occurred in five patients. PMID- 3027852 TI - Tolerance to and efficacy of itraconazole in treatment of systemic mycoses: preliminary results. AB - Twelve patients with mycotic infections were treated with itraconazole during a one-year period. Patients had the following conditions: coccidioidomycosis (three), histoplasmosis (three), mucocutaneous candidosis (two), aspergillosis (three), and urinary tract infection due to Torulopsis glabrata (one). The daily dose of itraconazole ranged from 100 to 200 mg. Ten of the 12 cases were assessable. Clinical improvement was observed in six patients, two with coccidioidomycosis, two with mucocutaneous candidosis, one with histoplasmosis, and one with aspergilloma. Four patients did not respond to therapy; two of these patients had aspergillosis, one had histoplasmosis, and one had T. glabrata infection. No adverse effects were attributable to itraconazole. PMID- 3027854 TI - [Intrapulmonary chemodectoma. A new case with ultrastructural study]. AB - The authors report the case of a 40-year old woman who presented with a round tumour in the middle lobe of the right lung. Once removed, the tumour proved to be an intrapulmonary chemodectoma. Chemodectomas are tumours that are rarely present in the lung. Their pathogenesis is open to discussion. Their study by electron microscopy is interesting, as it shows grains of neurosecretion and provides additional information on the morphology of chemoreceptors. PMID- 3027853 TI - Ketoconazole vs. itraconazole for antifungal prophylaxis in patients with severe granulocytopenia: preliminary results of two nonrandomized studies. AB - The efficacies of antifungal prophylaxis with ketoconazole and itraconazole, a new triazole, in patients with prolonged granulocytopenia were evaluated in two nonrandomized studies. The conditions other than the drug administered for prophylaxis were equivalent in the two studies. The incidence of fatal fungal infections was significantly higher among patients given ketoconazole than among those given itraconazole (P = .02); this trend was especially evident with fatal infections due to Aspergillus (P = .0045). The level of protection from fatal fungal infection afforded by itraconazole was highly significant among patients who were granulocytopenic for more than 25 days (P less than .0001) and among patients with acute lymphoblastic leukemia (P = .008). Failures of prophylaxis with itraconazole may have been due to inadequate levels of drug in plasma. Although these results must be interpreted with caution because they were obtained in consecutive nonrandomized studies, the use of itraconazole for antifungal prophylaxis in granulocytopenic patients seems to be a major advance and to be especially advantageous in hospitals, where the incidence of fatal aspergillosis in these patients is high. PMID- 3027855 TI - [A case of diffuse pulmonary amyloidosis associated with peripheral neurologic signs]. AB - The authors report a case of well-tolerated and prolonged diffuse parenchymal amyloidosis of the lungs associated with signs of peripheral neuropathy. The latest classifications of amyloidosis generally and of bronchopulmonary amyloidosis specifically are reviewed. The problems raised by this particular case, and notably its relation with the neurological signs observed, are discussed. PMID- 3027856 TI - [Current aspects of pulmonary granulomatosis]. PMID- 3027857 TI - [Mucoviscidosis (pathogenesis and evolution)]. PMID- 3027858 TI - [Loss of work capacity in chronic obstructive bronchopneumopathy]. PMID- 3027859 TI - [Epidemiologic, pathogenetic and clinico-therapeutic considerations in 23 cases of complications following BCG vaccination occurring in a group of young adults]. PMID- 3027861 TI - [Pathogenesis and surgical treatment of bullous emphysema]. PMID- 3027860 TI - [Polychemotherapy of small-cell bronchopulmonary cancer. The advantages of adding the selective cytostatic Riaval]. PMID- 3027862 TI - [Clinico-therapeutic considerations of a case of congenital pulmonary air cyst]. PMID- 3027864 TI - Variability of mycobacterial population in the process of the experimental chemotherapy of tuberculosis. PMID- 3027863 TI - [Segmental bronchoalveolar lavage with a flexible tube via a rigid bronchoscope in a case of mediastinopulmonary sarcoidosis]. PMID- 3027866 TI - [Transthoracic fine-needle aspiration biopsy in the diagnosis of tuberculoma]. PMID- 3027865 TI - The significance of L-forms of mycobacteria in the clinical picture of tuberculosis. PMID- 3027867 TI - [Evaluation of the integrated treatment of tuberculosis in 20 tuberculosis outpatient units]. PMID- 3027868 TI - [Pleurisy registered as active cases or as follow-up cases between 1974 and 1983]. PMID- 3027869 TI - [Follow-up assessment of extrarespiratory tuberculosis in the last 10 years (1975 1984)]. PMID- 3027871 TI - [Bacteriologic data on the isolation of Koch's bacillus from cases of extrapulmonary tuberculosis over an 8-year period]. PMID- 3027870 TI - [Results of short-term chemotherapy in Galati County 4 years after completion of treatment]. PMID- 3027872 TI - [Epidemiometric value of the tuberculin index for a group of 16,351 young adults]. PMID- 3027873 TI - Induction of stress proteins by hyperosmolarity in normal and transformed cells. AB - The synthesis of at least three proteins, with molecular weights of approximately 87, 70, and 53 kd, was enhanced following the exposure of chick embryo fibroblasts to hyperosmolar shock of 30 min at 0.6 osM. Two of these proteins, the 87 and 70 kd, comigrated on one-dimensional gel electrophoresis with the stress proteins induced by heat shock after 30 min at 44 degrees C. In 3T3 cells, the hyperosmolar shock enhanced the expression of two proteins of 88 and 52 kd, whereas the heat shock increased the synthesis of several new polypeptides including the 88 and 52 kd mw. In SV40-transformed 3T3 cells the synthesis of two proteins of 72 and 69 kd was enhanced by heat shock, but no change of the protein pattern was recorded after the hyperosmolar shock. PMID- 3027875 TI - [For clinical practice ...one should remember...]. PMID- 3027874 TI - [Hepatocellular carcinoma and alcoholic cirrhosis]. PMID- 3027876 TI - [Human papillomavirus infections of the female genital tract]. PMID- 3027877 TI - [Female genital virus infections, excluding papillomavirus infections]. PMID- 3027878 TI - Pseudomyopathic changes in peripheral neuropathies. PMID- 3027879 TI - Influence of cations on enamel phosphatase activity. AB - ATP and 5-Br-4-Cl-indoxyl phosphate were used as substrates for spectrophotometric measurement of phosphatase activity in partly mineralized bovine enamel. MgCl2, CaCl2, NaCl, and KCl were used as moderators of the enzyme activities. It is concluded that at least two phosphatases, active at alkaline pH, are present in the immature enamel matrix. PMID- 3027880 TI - High doses of fluoride do not affect cyclic AMP levels in human and rat plasma or urine. AB - The effect of fluoride ingestion on plasma and urinary cyclic AMP levels was studied in healthy volunteers, children undergoing prophylactic fluoride treatment and in rats. In the first series of experiments fluoride ingestion led to a 20-fold increase in plasma fluoride concentration, which was unrelated to changes in plasma cyclic AMP. The only significant effect on plasma cyclic AMP occurred prior to fluoride, an effect attributed to stress. In the second series performed in children increases in urinary F- of up to 200-fold were associated with slight but nonsignificant changes in cyclic AMP excretion. In rat experiments the blood sampling procedure was associated with a rise in plasma cyclic AMP. When this was prevented by pretreatment with propranolol (3 mg/kg), the effect of fluoride (50 ppm in the drinking water) was tested. A fall in urine production was not associated with any change in cyclic AMP excretion or in nephrogenic cyclic AMP. It is concluded that if fluoride alters cyclic AMP in rats and man the effect is small and easily masked by other factors such as catecholamine release following stress. PMID- 3027881 TI - Hormonal regulation of capillary fenestrae in the rat adrenal cortex: quantitative studies using objective lens staging scanning electron microscopy. AB - High magnification studies of the fenestrated capillary endothelium in the zona fasciculata (ZF) of rat adrenal glands were performed using the objective lens stage of an analytical scanning electron microscope (SEM) equipped with a lanthanum hexaboride emitter (LaB6). Resolution of surface substructure of the luminal membrane obtained with specimens decorated with gold/palladium (Au/Pd) was compared with that observed in others sputter coated with tantalum (Ta). High magnification (50,000x) of the fenestrated endothelium demonstrates that tantalum coating of the cryofractured adrenals improves the substructural detail compared to that seen in Au/Pd decorated specimens. The procedures used in specimen preparation, metal deposition and secondary electron imaging (SEI) are described. Quality imaging achieved using the objective lens stage is a result of the elimination of the SE-III component derived from backscattered electrons. Rat adrenals exhibited uniformly patent capillaries. High magnification micrographs of capillary walls were randomly recorded in two morphometric studies of the fenestral content of capillaries in the rat adrenal cortex. Adrenocorticotropic hormone (ACTH), when administered to rats following dexamethasone (DEX) treatment, significantly reduced the fenestrae/micron 2 of endothelial surface and increased the mean size of fenestrae. After hypophysectomy, the number of fenestrae/micron 2 declined over 48 h; within 2 h after ACTH was given to rats hypophysectomized 48 hours earlier, the fenestrae/micron 2 had increased two fold. These studies indicate that ACTH plays an important role in modulating fenestral content of the capillary endothelium in the adrenal cortex. PMID- 3027882 TI - The motile behavior of virus-transformed 3T3 cells. AB - Using metallic gold in various assays for the motility of cultured tissue cells, the paper compares the movements of surface projections and the locomotion of polyoma (Py3T3) and SV40 (SV3T3) virus-transformed 3T3 cells with the behavior of the parental 3T3 cells. The movement of surface projections was assayed by the ability of filopodia, lamellipodia and blebs of freshly plated cells to remove colloidal gold particles from a particle-coated glass substrate. The ability of filopodia to probe the environment for points of anchorage was tested by observing cells plated on glass whose filopodia touched the surface of a neighboring area of evaporated gold. The locomotion of cells was assayed by particle-free tracks (phagokinetic tracks) which were left by migrating cells on a glass substrate which was previously coated with colloidal gold particles. The paper suggests that the ability of the transformed cells to sense environmental factors, and their behavioral controls are altered. PMID- 3027883 TI - Distinct histamine-induced cyclic AMP synthesis in acute leukemia. AB - Histamine receptors of the H2 type are found on mature hemopoetic cells such as lymphocytes and monocytes. Since little is known about histamine receptors on immature cells, we studied histamine-induced cAMP synthesis in leukemic cells of 51 patients using 73 samples of peripheral blood and/or bone marrow. Histamine induced cAMP synthesis was found in all cALL (FAB L1, L2), but only occasionally in AML. In 1 out of 11 myelomonocytic leukemias (M4), and also in 2 out of 5 monocytic leukemias (FAB M5), histamine-inducible cAMP synthesis was found, but no induction of cAMP synthesis by histamine was found in 19 cases from the remaining subgroups (FAB M1-M3, M6). The inhibiting action of H2 antagonists on cAMP synthesis indicates that the receptor is of the H2 type. Since H2 receptors were found in all cALL blasts, we assume a compulsory expression of this receptor on immature cells of the B-lymphocytic lineage. PMID- 3027884 TI - Evidence of tinidazole interference in the oropharyngeal inflammatory process during infectious mononucleosis. AB - The efficacy of tinidazole was studied in 24 patients with infectious mononucleosis (IM), 13 of whom were randomized for 5 days of tinidazole treatment and 11 for control without placebo. In judging the comparability of the 2 groups not only was the distribution of confounding factors such as age, sex or duration of symptoms before admission considered, but also the distribution of the Epstein Barr virus (EBV) serological stage at entry. In these respects the groups were well matched. The duration of sore throat and pharyngotonsillitis after admission was significantly shorter for the patients treated with tinidazole than for the controls. Orosomucoid and lactodehydrogenase concentrations normalized more readily in IM patients with a short duration of sore throat after admission and in patients treated with tinidazole compared to those with a long duration of sore throat and to the tinidazole controls. Clinical and laboratory findings were thus parallel and showed a clinical effect of tinidazole, believed to be mediated via the well-known activity of this drug against anaerobic bacteria. The EBV serological stage of each patient at entry could not predict the duration of symptoms. The results showed that the course of the angina was not primarily dependent on the virus host interaction but probably on other factors, still unknown. One such factor could be the balance between the mucosal defence and the normal oropharyngeal microflora. PMID- 3027885 TI - Innervation of the child urinary bladder. AB - Muscle strips from the fundus, trigonum and distal ureters obtained from children at operations for vesico-ureteric reflux were studied using histochemical and immunohistochemical methods, and electrical nerve stimulation in an organ bath. A rich supply of cholinergic nerves was found and despite a partial atropine resistance the neurophysiological experiments indicated that the transmitter causing contraction of the detrusor muscle is acetylcholine. Imipramine, which is used in the treatment of enuresis, had no anticholinergic effect on the bladder in the doses used clinically. The adrenergic innervation was very sparse except around the ureter orifices. No contractile alpha-adrenoceptors could be detected but beta receptor mediated relaxation was found, which was neither of the beta 1 nor beta 2 type. A third type of beta receptor is postulated. Peptidergic nerves containing vasoactive intestinal peptide, VIP, were demonstrated in a few nerve terminals running along bundles of smooth muscle. No nerves containing enkephaline, somatostatine or substance P were found. VIP affected the detrusor muscle indicating a possible role as a modulator of transmitter action, while substance P had no effect. The anticholinergic and calcium antagonistic drug terodiline inhibited all muscle activity, whether it was induced by nerve stimulation, cholinergic drugs or a potassium rich solution, making it suitable for treatment of diurnal enuresis. PMID- 3027886 TI - Sex education and rehabilitation with schizophrenic male outpatients. AB - Research indicates that schizophrenic patients lack intimate relationships and show a high rate of sexual dysfunction. Despite increasing awareness of the rights of handicapped persons to sexual expression, the treatment of schizophrenic patients rarely addresses their sexuality. A sex education program for recent-onset male schizophrenic patients attending an outpatient clinic was developed in response to several incidents involving patients' inappropriate sexual behaviors. To enhance our understanding of the current sexual functioning and needs of these patients, sex histories were taken. Almost all of the 16 patients interviewed were sexually active, with autoerotic activity predominating. Sixty-three percent of the patients reported orgasmic and/or erectile dysfunctions. Other studies have linked sexual dysfunction to the side effects of antipsychotic medications. The objectives of the sex education program were: to provide information; to clarify values; to overcome sexual dysfunction; and to enhance intimacy skills. The authors used role playing, modeling, group exercises, and explicit sex therapy audiovisual material to improve patients' intimacy skills. Patients participated actively and used the group to explore sexual issues. No exacerbations of symptoms were observed among patients participating in the program. PMID- 3027887 TI - Large granular-cell myoblastoma of the oesophagus. AB - A granular-cell tumour of the oesophagus, the largest hitherto described, was found in a 44-year-old woman. Histologic examination showed an infiltratively growing granular-cell myoblastoma without pleomorphism or mitotic activity. Local extirpation of the tumour was attempted in order to preserve oesophageal continuity. However, a tracheo-oesophageal fistula and stenosis of the previously tumour-bearing area developed and necessitated nasogastric tube feeding for 18 months. Two attempts to close the fistula, including resection of the fistula bearing tracheal area, failed. Final cure was achieved by subtotal extirpation of the oesophagus and gastro-oesophageal anastomosis in the neck with the stomach placed retrosternally. Normal intake of food was restored after this operation and 30 months later the patient is doing well. PMID- 3027888 TI - [Pathology of primary lung cancers in cattle (description of 16 cases)]. PMID- 3027889 TI - Coliphage M13 cloning system and ddXTP chain-termination method for DNA sequencing. AB - Two DNA fragments of cytochrome b gene in yeast mitochondrial DNA were sequenced by Messing's M13 cloning system and Sanger's ddXTP chain-termination method. M13mp8 and M13mp9 serve as vectors for insert fragment. The recipient strain is E. coli JM103 for transfection. When the ratio of insert/vector was 3:1, high frequencies of recombination and positive recombination were obtained. The two fragments, which have 575 bp and 709 bp, were sequenced. To read more bases, the ratio of ddXTP/dXTP must be reduced. On one X-ray film of 80 x 17 cm 394 bp were read. PMID- 3027890 TI - The expression of HBsAg gene in yeast under GAL-10 promoter control. AB - The surface antigen gene of HBV (adr subtype) is inserted into the YEP 1 which is replicated and selected in E. coli and baker yeast Saccharomyces cerevisiae. The HBsAg gene can be expressed in yeast under GAL-10 promoter control. The protein synthesized in yeast is assembled into particles that have the same size and shape as particles isolated from plasma. One microgram surface antigen can be yielded in 100 ml culture. PMID- 3027891 TI - The biology and chemistry of fertilization. AB - Fertilization of eggs by sperm, the means by which sexual reproduction takes place in nearly all multicellular organisms, is fundamental to the maintenance of life. In both mammals and nonmammals, the pathway that leads to fusion of an egg with a single sperm consists of many steps that occur in a compulsory order. These steps include species-specific cellular recognition, intracellular and intercellular membrane fusions, and enzyme-catalyzed modifications of cellular investments. In several instances, the molecular mechanisms that underlie these events during mammalian fertilization are beginning to be revealed. PMID- 3027892 TI - The mitochondrial genotype can influence nuclear gene expression in yeast. AB - Isochromosomal, respiratory-deficient yeast strains, such as a mit-, a hypersuppressive petite, and a petite lacking mitochondrial DNA, are phenotypically identical in spite of differences in their mitochondrial genomes. Subtractive hybridizations of complementary DNA's to polyadenylated RNA isolated from derepressed cultures of these strains reveal the presence of nuclear-encoded transcripts whose abundance varies not only between them and their respiratory competent parent, but among the respiratory-deficient strains themselves. Transcripts of some nuclear-encoded mitochondrial proteins, like cytochrome c and the alpha and beta subunits of the mitochondrial adenosine triphosphatase, whose abundance is affected by glucose or heme, do not vary. In the absence of major metabolic variables, yeast cells seem to respond to the quality and quantity of mitochondrial DNA and modulate the levels of nuclear-encoded RNA's, perhaps as a means of intergenomic regulation. PMID- 3027893 TI - Drug may protect brains of heart attack victims. PMID- 3027894 TI - HTLV x gene mutants exhibit novel transcriptional regulatory phenotypes. AB - The human T-cell leukemia viruses, HTLV-I and HTLV-II, contain a gene, termed x, with transcriptional regulatory function. The properties of the x proteins were analyzed by constructing mutant genes containing site-directed deletions and point mutations. The results demonstrate that the amino terminal 17 amino acids of the x protein constitute part of a functional domain that is critical for the transcriptional activating properties of the protein. Within this region, substitution of a leucine residue for a proline residue results in major changes in the trans-activation phenotype of the protein. The mutant HTLV-II x protein, though incapable of activating the HTLV-II long terminal repeat, will block trans activation of the HTLV-II long terminal repeat by the wild-type protein. The altered phenotype of this mutant suggests a potential negative regulatory function of the x protein. PMID- 3027895 TI - Synthesis of a site-specific DNA-binding peptide. AB - The Hin recombinase binds to specific sites on DNA and mediates a recombination event that results in DNA inversion. In order to define the DNA-binding domain of the Hin protein two peptides 31 and 52 amino acids long were synthesized. Even though the 31mer encompassed the sequence encoding the putative helix-coil-helix binding domain, it was not sufficient for binding to the 26-base pair DNA crossover site. However, the 52mer specifically interacted with the site and also effectively inhibited the Hin-mediated recombination reaction. The 52mer bound effectively to both the 26-base pair complete site and to a 14-base pair "half site." Nuclease and chemical protection studies with the 52mer helped to define the DNA base pairs that contributed to the specificity of binding. The synthetic peptide provides opportunities for new approaches to the study of the nature of protein-DNA interaction. PMID- 3027896 TI - U3 sequences from HTLV-I and -II LTRs confer pX protein response to a murine leukemia virus LTR. AB - Human T-cell leukemia virus (HTLV) types I and II are unusual among replication competent retroviruses in that they contain a fourth gene (chi) necessary for replication. The chi gene product, p chi, transcriptionally transactivates the viral long repeat (LTR), and is thus a positive regulator. To investigate p chi transactivation, sequences from the U3 regions of the LTRs of HTLV-I and -II were inserted into the Moloney murine leukemia virus (M-MuLV) LTR by recombinant DNA techniques. Transient expression assays of the chimeric LTRs indicated that the HTLV sequences conferred to the M-MuLV LTR responsiveness to HTLV p chi protein. M-MuLV enhancers were not required for function of the chimeric LTRs. Infectious recombinant M-MuLVs containing chimeric LTRs were also generated. These viruses showed higher infectivity when assayed in mouse cells expressing HTLV-II p chi protein compared to normal mouse cells. Thus the HTLV sequences were able to confer p chi responsiveness to infectious M-MuLV. The generation of a virus dependent on a transactivating protein for its replication has implications for the evolution of the human T-cell leukemia viruses. PMID- 3027898 TI - [Use of correlation coefficients of peripheral blood enzymes in endogenous infections]. PMID- 3027899 TI - Fusion to the sacrum for nonparalytic scoliosis in the adult. AB - This study is a retrospective review of 43 adult patients with idiopathic or congenital scoliosis who had spinal fusion from T11 or above to the sacrum. This study was prompted by the frustrations of the treating surgeons in attempting long fusions from the thoracic spine to the sacrum. Of 25 patients treated with a single-stage posterior fusion only 28% had a good result with a single procedure. Failures were due to pseudarthrosis, decompensation, or loss of lumbar lordosis. Ten patients treated with posterior fusion and subsequent 6-month augmentation had a 70% success rate. Eight patients treated with anterior followed by posterior fusion had a 75% success rate. The ideal answer to this clinical problem has not yet been found. PMID- 3027897 TI - Case report 403: Extra-mammary Paget disease of the skin with disseminated skeletal metastases. PMID- 3027901 TI - Human gastrin-releasing peptide gene maps to chromosome band 18q21. AB - A complementary DNA clone encoding human pre-pro gastrin-releasing peptide, a 27 amino acid neuropeptide and putative growth factor, was used to determine the chromosomal location of this gene. Southern blot hybridization to genomic DNA isolated from a panel of human-rodent somatic cell hybrids unambiguously maps this gene to human chromosome 18. In situ chromosomal hybridization confirms the hybrid data and further localized the gene to chromosome band 18q21. Karyotypic abnormalities in tumors and inherited disease states which involve chromosome band 18q21 may now be studied for correlated changes in the structure and expression of the human GRP gene. PMID- 3027900 TI - Analysis of spontaneous mutations in a chromosomally located HSV-1 thymidine kinase (TK) gene in a human cell line. AB - We have developed a system for studying spontaneous mutations at a chromosomally located single-copy HSV-1 thymidine kinase (TK) gene in the human 143 TK- cell line. The neo gene, which confers resistance to the antibiotic G418, was placed next to the TK gene for the purpose of screening out gross chromosomal alterations. TK- mutations were selected using the anti-TK nucleotide analogs trifluorothymidine, acyclovir, and DHPG 9-(1,3 dihydroxy-2-propoxymethyl)-guanine either separately, or in combination to eliminate leaky mutations. Analysis of the TK- mutations by Southern blotting revealed that the majority had undetectable alterations of less than 50 base pairs. The results using the methylation-sensitive enzymes HpaII, AvaI, and SmaI suggest that the inactivation of the TK gene was not due to extensive methylation, although specific methylation of a limited number of MspI sites cannot be ruled out. Reversion studies, however, showed that of 16 mutants analyzed, about half had a very high reversion frequency (approximately 10(-2). This suggests that inactivation of the TK gene may have occurred by a variety of mutational events. PMID- 3027902 TI - Human gastrin-releasing peptide gene is located on chromosome 18. AB - Gastrin-releasing peptide (GRP), a bombesin-like peptide, increases plasma levels of gastrin, pancreatic polypeptide, glucagon, gastric inhibitory peptide, and insulin. GRP is produced in large quantities by small-cell lung cancer and acts as a growth factor for these cells. To determine if chromosomal changes in small cell lung cancer are related to the expression of GRP, we chromosomally mapped the gene using human-mouse somatic cell hybrids. Twenty hybrids, characterized for human chromosomes, were analyzed by Southern filter hybridization of DNA digested with EcoRI. Human DNA cut with EcoRI yields a major band of 6.8 kb and a minor band of 11.3 kb. The 6.8 kb band segregated concordantly with chromosome 18 and the marker peptidase A. The chromosome 3 abnormalities seen in small-cell lung cancer do not correlate with the chromosomal location of GRP, suggesting that the elevated expression of this gene may be due to mechanisms other than chromosomal rearrangement. PMID- 3027903 TI - Assignment of mouse beta-spectrin gene to chromosome 12. AB - The structural gene for the beta-subunit of the mouse erythrocyte spectrin, hereinafter designated as Sp-b, was assigned to the mouse chromosome 12. This assignment was made by Southern analysis of genomic DNA from mouse X Chinese hamster hybrid cells using cloned mouse erythrocyte beta-spectrin cDNA as a probe. In the PstI-digested genomic hamster cell DNA a single band of 2.0 kb was detected, whereas PstI-digested mouse DNA gave a band of 4.2 kb, when probed with the mouse erythroid beta-spectrin cDNA clone. This allowed us to analyze a panel of mouse X Chinese hamster somatic cell hybrids to map this gene to chromosome 12. Interestingly, this assignment is different from that observed for the alpha subunit of spectrin, which has been mapped to chromosome 1 in mouse. These results serve as a basis for further genetic characterization of the mouse hemolytic anemias. PMID- 3027904 TI - [Neuroectodermal melanotic tumor of the upper jaw in childhood]. PMID- 3027905 TI - A comparison of lisinopril and atenolol in black and Indian patients with mild-to moderate essential hypertension. AB - The effects of lisinopril (MK-521; MSD) and atenolol in the treatment of mild-to moderate essential hypertension were compared in a double-blind, parallel, controlled study, with 24 patients randomly assigned to lisinopril and 12 to atenolol. Patients in both groups whose blood pressure did not respond satisfactorily were given hydrochlorothiazide (HCTZ). The groups were matched for age, race and pretreatment blood pressure. Response to therapy was similar in the atenolol and lisinopril groups, but the combination of lisinopril and HCTZ produced a better response than atenolol plus HCTZ. Indian patients appeared to respond better to lisinopril than black patients, but the hypotensive response to lisinopril plus HCTZ was similar. Plasma renin activity rose and the plasma aldosterone level fell after taking lisinopril. Adverse effects did not occur and only 1 patient on lisinopril 20 mg/d was withdrawn from the study because of postural hypotension. Lisinopril alone and combined with HCTZ in Indian patients and lisinopril combined with HCTZ in black patients produced a satisfactory fall in blood pressure. PMID- 3027906 TI - [Enzymes of purine metabolism in thymic subpopulations defined by lectins]. PMID- 3027908 TI - Solitary cerebral metastasis from Wilms' tumor without pulmonary involvement. AB - A 6-year-old boy presented with a solitary cerebral metastasis 5 years after nephrectomy for stage I Wilms' tumor. Chest x-ray and computed tomography scan were normal. The occurrence of cerebral metastasis without pulmonary involvement in Wilms' tumor has not been described before. PMID- 3027907 TI - Immunopathogenesis of the neuropsychiatric manifestations of systemic lupus erythematosus. PMID- 3027911 TI - Modified quadruple-loop (W) ileal reservoir for restorative proctocolectomy. AB - Reconstructive ileal pouch procedures are well-accepted alternatives to a permanent ileostomy in select patients requiring proctocolectomy for ulcerative colitis and familial polyposis. Double-, triple-, and quadruple-loop and lateral isoperistaltic designs have been described; however, the ideal pouch capacity and configuration are still debatable. Evidence has been reported that improved functional results and lower stool frequency with less antidiarrheal medication can be achieved with larger volume reservoirs. We describe a new modification of a quadruple-loop pouch that has been successfully performed in 10 patients. This new approach to pouch construction produced excellent functional results 6 to 12 months after ileostomy takedown and was easier to construct and engage into the muscular cuff of the distal rectum than quadruple-loop pouches of equal length loops. PMID- 3027909 TI - [Fundamental model assays on chemical damages to ribonucleic acids caused by oxygen radicals in aqueous solution]. AB - A partial decomposition of RNA molecules in Saccharomyces cerevisiae is caused by photochemically produced superoxide anion radical (O2-.). Hydroxyl radicals (OH.) lead to complete destruction of RNA. Such radicals as originating from peroxidase hydrogen peroxide reaction cause a partial decomposition of RNA molecules. All RNA species are damaged to the same extent by radicals. Damages due to oxygen radicals can be prevented or reduced by strong radical captors (dimethyl sulfoxide, 1,4-diazabicyclo-(2,2,2)-octane or superoxide dismutase). All these reactions proceed in aqueous solution. Correlations with the use of radicals in radiotherapy are discussed. PMID- 3027910 TI - The interaction of retrovirus and chemical carcinogen in experimental colon carcinogenesis. AB - The isolation and characterization of oncogenes from human colon cancer and the recognition of their homology with the ras gene of the Harvey and Kirsten strain of murine sarcoma virus (MSV) led us to investigate the effect of exogenous MSV on 1,2 dimethylhydrazine (DMH)-induced colon carcinoma in rats. DMH, 20 mg base/kg, was injected weekly for 10 weeks into Sprague-Dawley rats. The Moloney murine sarcoma virus (MSV-M) was injected (200 focus-forming units) intraperitoneally into 15 rats 48 hours after the last DMH injection or in 12 rats before the first DMH injection. Controls consisted of 12 rats receiving 10 injections of DMH only, nine rats receiving MSV-M alone, and 10 untreated rats. All tumors induced were adenocarcinomas of the gastrointestinal tract, characteristically induced by DMH and not by MSV-M. In the late virus group there was an augmentation of colon tumor induction (mean, 2.2 versus 1.1 colon tumors/rat, p less than 0.05), and in the MSV pretreated group, there was also significant augmentation of colon tumor induction (mean, 2.4 versus 1.1 colon tumors/rat, p less than 0.005) when compared with rats treated with DMH alone. Rats treated with MSV-M alone and untreated rats had no tumors. This is the first study to suggest the importance of exogenous viral infection in chemically induced colonic carcinogenesis. PMID- 3027912 TI - [Cough induced by converting enzyme inhibitors]. PMID- 3027913 TI - [Angioedema after administration of enalapril in a chronic renal failure patient]. PMID- 3027914 TI - Control of thrombin mediated cleavage of protein S. AB - Thrombin has been shown to cleave the vitamin K dependent cofactor protein S with subsequent loss of its cofactor activity. This study examines the control mechanisms for thrombin cleavage of protein S. The anticoagulant activity of activated protein C (APC) is enhanced fourteen fold by the addition of protein S. Thrombin cleaved protein S is seven fold less efficient than the native protein, and this loss of activity is due to reduced affinity of cleaved protein S for APC or the lipid surface compared to the intact protein. In the absence of Ca++, protein S is very sensitive to minimal concentrations of thrombin. As little as 1.5 nM thrombin results in complete cleavage of 20 nM protein S in 10 min and loss of cofactor activity. Ca++, in concentrations greater than 0.5 mM, will inhibit this cleavage and in the presence of physiological Ca++ concentrations, no cleavage of protein S could be demonstrated in spite of high concentrations of thrombin (up to 1 microM) and prolonged incubations (up to two hours). The endothelial surface protein thrombomodulin is very efficient in inhibiting the cleavage of protein S by thrombin suggesting that any thrombin formed on the endothelial cell surface is unlikely to cleave protein S, thus allowing the intact protein to act as a cofactor to APC. We conclude that the inhibitory effects of Ca++ and thrombomodulin on thrombin mediated cleavage of protein S imply that this event, by itself, is unlikely to represent a physiological control of the activity of protein S. PMID- 3027915 TI - [Dietary fiber tablets and intestinal function]. PMID- 3027916 TI - [Polyneuropathy after pure petrol sniffing]. PMID- 3027917 TI - [BVD (bovine virus diarrhea) and IBR (infectious bovine rhinotracheitis): serologic studies on 35 farms]. AB - 1099 female cattle from 35 farms in the provinces of North Brabant, Gelderland and Overijssel were examined for the presence of antibodies against Bovine Virus Diarrhoea (BVD) and Infectious Bovine Rhinotracheitis (IBR) by serum neutralisation tests. One of the farms examined was seronegative for BVD and 11 farms were seronegative for IBR. On the seropositive farms about 50% of cattle carrying their first calf had not experienced infection i.e. were seronegative (BVD 51.2%, IBR 48.3%). After calving the percentages of seronegative cows were considerably lower (BVD 19.6%, IBR 28.6%). The cause, implications, and prospects of prevention (vaccination) of the diseases are discussed. PMID- 3027918 TI - In vivo interaction of antibodies with cell surface proteins used as antigens. AB - Antibody interactions with cell membrane glycoproteins in vivo exhibit features of aggregation and capping with resultant shedding similar to those events described in several in vitro isolated cell systems. Requirements for divalent ligand binding and participation of cytoskeletal elements are demonstrated in vivo as well. Persistence of antigen in immune complexes with complement interaction in situ appear to be necessary to induce an inflammatory response in vivo. Abrogation of this response occurs when circumstances permit antigenic modulation with disappearance of the immune complex. PMID- 3027919 TI - Corticosteroid binding in human artery. AB - Mineralo- and glucocorticoid binding in cytosol of surgically obtained human arteries and umbilical arteries were analyzed. Scatchard analysis of glucocorticoid binding in femoral and umbilical arteries revealed the binding parameters of Kd = 4.3, 8.2 X 10(-9) M; Bmax = 42, 90 fmol/mg protein, respectively. Scatchard analysis of mineralocorticoid binding in inferior mesenteric artery showed a curvilinear pattern suggestive of the presence of two binding components, one with a higher affinity and a lower capacity for the ligand (Kd = 3.3 X 10(-10) M, Bmax = 0.8 fmol/mg protein) and the other with lower affinity and higher capacity (Kd = 10 X 10(-7) M, Bmax = 36 fmol/mg protein). Polyacrylamide gel electrophoresis of mineralo- and glucocorticoid binding in the cytosol of popliteal artery also showed the presence of the corticosteroid binding components with limited binding capacity. Sucrose density gradient centrifugation of corticosteroid binding in cytosol of femoral artery revealed the presence of dexamethasone binding molecule sedimenting in the 8-9 S region and deoxycorticosterone binding molecules sedimenting in the 7-8 and 8-9 S regions of the gradinent. Those results revealed the presence of mineralo- and glucocorticoid receptors in human artery and suggest the possible direct actions of corticosteroids upon human artery. PMID- 3027920 TI - Studies on the effect of chronic L-triiodothyronine (T3) treatment on brain Na+,K+-ATPase activity in the mature rat. AB - Mature rats were dosed with T3 by different routes and dose-levels at either 0.1 mg/kg for 14 days s.c. (Group A), 1 mg/kg for 3 alternative days i.p. (Group B), 5 mg/kg for 14 days p.o. (Group C), or with propylthiouracil (PTU 50 mg/day for 14 days p.o.-Group D). Measurement of cerebellar and striatal NA+,K+-ATPase activities showed that whereas Groups A, B and D were unaffected when compared with controls, there were 35-70% increases respectively in the activities of both molecular forms of the enzyme, alpha(+), high ouabain affinity, and alpha, low ouabain affinity, in Group C rat brains at the highest dose of T3 tested. Kidney Na+,K+-ATPase activity was also elevated (67% increase) in this group of animals showing significant changes in renal medullary tissue only. Acute elevation of brain dopamine levels by administration of an MAOI plus L-DOPA (50 mg/kg, 60 min) significantly elevated (20% increase) the activities of both molecular forms of Na+,K+-ATPase in corpus striatum. Treatment with L-tryptophan (50 mg/kg, 60 min) failed to produce any changes in the striatal activities. The possible relationship of increases in enzyme activities with T3 and increased brain monoamine function is discussed. Both plasma free T4(FT4) and total T4(TT4) were markedly depressed in all T3-treated rats. Although hypothalamic thyrotropin releasing hormone (TRH) concentrations were unaltered by any of the T3 treatments, pituitary thyroid stimulating hormone (TSH) concentrations were greatly diminished and it is thought that this may reflect a direct effect of T3 on TSH synthesis. PMID- 3027922 TI - Subcellular fractionation of Trypanosoma cruzi; isolation and characterization of plasma membranes from epimastigotes. AB - A procedure is described for the isolation of subcellular fractions from epimastigotes of Trypanosoma cruzi. The method could separate most of the nuclei, mitochondria and microsomes. The plasma membranes were purified by discontinuous density gradient centrifugation in alkaline buffer containing sucrose and magnesium. The yield of plasma membrane was 3.7 mg of protein per 10(9) epimastigotes, accounting for approximately 4.2% of the total cell proteins. The plasma membrane obtained from the 34-50% interface of sucrose density gradients was shown, by electron microscopy, to be completely free of other organella and was homogeneous according to enzymatic marker criteria. The specific activity of the 5'-nucleotidase and acid phosphatase were 96- and 5.5-fold, respectively, higher than that in the total cells, suggesting that the enzymes can be considered as good plasma membrane markers for T. cruzi. The results confirmed the possibility of the presence of membrane-bound acid phosphatase of T. cruzi. PMID- 3027921 TI - Mechanism of action of the platelet function inhibitor from Vipera russelli siamensis snake venom. AB - Human platelet aggregation induced by ADP, adrenaline, collagen or thrombin was inhibited by the venom inhibitor. Heating reduced both its phospholipase A2 enzymatic and anti-aggregatory activities, although not in parallel. The inhibitor caused significant dose-related inhibitory effects on the clot retraction of rabbit platelet-rich plasma caused by thrombin, while platelet malondialdehyde formation stimulated by thrombin was not affected. Furthermore, the venom inhibitor increased basal cyclic AMP levels in platelets, while cyclic GMP content was slightly lowered, but not in a dose-dependent manner. In addition, microscopic study revealed that the cytoskeleton was disordered after treatment of platelets with the venom inhibitor. The platelets lost their discoid form, while the ultrastructural changes of platelet aggregation induced by ADP were blocked. It is concluded that increasing platelet cyclic AMP and the disorder of the cytoskeleton may be the mechanism of action of the venom inhibitor on platelet function. PMID- 3027923 TI - Syringocystadenoma papilliferum developed from giant comedo: a case report. AB - A case of syringocystadenoma papilliferum in a 44-year-old Japanese male provided interesting clinical and histological findings. The lesion was diagnosed clinically as an infectious when on the flexor aspect of the right thigh. Optical microscopical examination of serial sections showed that the tumor parenchyma was deeply seated along the boundary zone between the cutis and subcutis around the lower half of a giant comedo, and extended into the surrounding tissues from the non-keratinized epithelial portion of the giant comedo where the non-keratinized epithelium merged with the upper, keratinized epithelium. The tumor thus appeared to have developed from the non-keratinized transitional epithelium corresponding to the intrafollicular part of the apocrine sweat gland apparatus. Although 103 cases of syringocystadenoma papilliferum have been reported from Japan since 1924, the present findings are both clinically and histologically unique. PMID- 3027924 TI - Effect of allopurinol on ischemia and reperfusion-induced cerebral injury in spontaneously hypertensive rats. AB - In spontaneously hypertensive rats, we studied the participation of xanthine oxidase-linked free radical in ischemia and reperfusion-induced cerebral injury, using allopurinol, a xanthine oxidase inhibitor. The loss of righting reflex was noted in some animals after a 4 hour occlusion of bilateral common carotid arteries and 19 of 25 animals died within 72 hours after reperfusion. One hour after reperfusion, the cerebral water content increased significantly, with an increase in sodium content and a decrease in potassium content. In 7 animals treated with oral administrations of allopurinol (200 mg/kg) 24 hours and 1 hour before occlusion, no death was found either during occlusion or after reperfusion, and the loss of righting reflex was noted in only one animal 24-72 hours following reperfusion. The increase in cerebral water content and accompanied changes in electrolyte contents were clearly prevented by allopurinol. These results suggest the possibility that the production of xanthine oxidase-linked free radical participates in cerebral injury due to ischemia and reperfusion in spontaneously hypertensive rats. PMID- 3027925 TI - Superoxide anion radical does not mediate vasodilation of cerebral arterioles by vasoactive intestinal polypeptide. AB - We examined the hypothesis that oxygen radicals may mediate the vasodilator effect of VIP on cerebral arterioles in cats equipped with cranial windows. The appearance of superoxide anion radical in cerebral extracellular space during VIP application was examined by measuring the rate of superoxide dismutase (SOD) inhibitable reduction of nitroblue tetrazolium (NBT). Although VIP (1 and 10 micrograms/ml) caused substantial reduction of NBT, the rate of the SOD inhibitable portion was not significantly different from zero. We also examined the effect of scavenging of superoxide and hydrogen peroxide by topical application of SOD plus catalase on the vasodilator effect of VIP (0.05-1.0 microgram/ml). The dilation in response to VIP was not significantly affected in either large or small arterioles by scavenging of superoxide and hydrogen peroxide. We conclude that VIP does not cause generation of superoxide and that superoxide or other reactive oxygen species derived from it, such as hydrogen peroxide and hydroxyl radical, are not mediators of the cerebral vasodilation caused by VIP. PMID- 3027927 TI - The effect of graded hypoxia on the hippocampal slice: an in vitro model of the ischemic penumbra. AB - Submerged hippocampal slices were exposed to 30 minutes of moderate or mild hypoxia at 29 degrees C and then reoxygenated. Synaptic transmission was lost at the same rate in response to either grade of hypoxia, but recovery was faster following mild hypoxia. Hyperexcitability of synaptic transmission was a lasting feature following moderate hypoxia, but it was transient following mild hypoxia; after mild hypoxia the strength of synaptic transmission eventually returned to normal. Extracellular calcium did not change during moderate hypoxia. The extracellular pH of slices was always more acid than the bath; pH decreased further in response to both moderate and mild hypoxia. Extracellular potassium increased more during moderate than during mild hypoxia, and a period of rapid potassium uptake was also more pronounced following moderate hypoxia. Extracellular DC potential demonstrated a small positive shift during hypoxia, more so during mild hypoxia. These experiments suggest that synaptic function can be reversibly suppressed in mildly hypoxic brain tissue without severe depolarization of neurons; in addition, the degree and duration of posthypoxic hyperexcitability are correlated with the degree of hypoxia and the magnitude of the release of K+ from cells into their environment. PMID- 3027926 TI - Gangliosides (GM1 and AGF2) reduce mortality due to ischemia: protection of membrane function. AB - As evidenced by their ability to reduce cerebral edema, exogenous ganglioside administration exerts acute effects on CNS injury processes. We report here that ganglioside (GM1 or AGF2) treatment results in a 52% decrease in mortality 48 hours after the induction of ischemia in gerbils by permanent unilateral ligation of the common carotid artery. By comparing the occluded vs. nonoccluded sides of the brain (cortex and hippocampus) we found a significant loss of membrane Na, K ATPase activity due to ischemia in control animals, but no such differences were found between the hemispheres of ganglioside-treated gerbils. We hypothesize that gangliosides may be "protecting" membrane function as indicated by these ATPase analyses, reducing local CNS damage at the time of injury (i.e., reduced cell loss, fiber degeneration, membrane failure). By acutely limiting the extent of CNS tissue damage, conditions may be optimized for any subsequent CNS regrowth and functional recovery. PMID- 3027928 TI - [Electron microscopy study of the dissolution of the hydroxyapatite crystals of enamel treated with phosphorus acid]. PMID- 3027929 TI - Harmful effect of high stibogluconate treatment of kala-azar in India. PMID- 3027930 TI - Incidence of cytomegalovirus disease in cyclosporine-treated renal transplant recipients based on donor/recipient pretransplant immunity. AB - We retrospectively reviewed the clinical data of all renal transplant patients treated with cyclosporine as their main chronic immunosuppressive agent between 12/83 and 11/85 to identify cytomegalovirus-negative patients at our institutions who received cytomegalovirus (CMV)-positive kidneys. Using a latex agglutination test, twenty-two such patients were identified, of whom 2 were excluded due to early death and lack of posttransplant follow-up serology. Of the remaining 20 patients, 12 developed CMV antibody in the first 4 months posttransplant, and of these, 11 were hospitalized with complications related to primary CMV disease. Two of these seroconverting patients eventually died, and one lost her kidney. Of the 8 persistantly CMV-negative patients, 1 lost his kidney soon after transplantation, and one had a febrile illness 4 months posttransplant caused by a bacterial pneumonia. Concomitantly, 145 renal transplants (CMV-negative recipient receiving a CMV-negative kidney or CMV-positive recipient receiving either positive or negative kidneys) given to 142 patients functioned for at least 4 weeks. Only 3 cases of CMV reactivation disease occurred in previously antibody-positive patients. We conclude that the transplantation of a cytomegalovirus-positive kidney into a CMV-negative recipient carries a high risk of mortality/morbidity from primary cytomegalovirus disease. On the other hand, reactivation of CMV disease was uncommon early in the posttransplant course of cyclosporine-treated patients. PMID- 3027931 TI - [Uptake of cyclic adenosine-3,5-monophosphate by cultured mammalian cells and frog muscles]. AB - Uptake of 3H-cAMP by isolated frog sartorii muscles and cultured mouse 3T3 and 3T6 cells was studied. It was shown that after 1-2 hours of incubation the intracellular level of cAMP in frog muscles reached 10-20% of its concentration in the incubation medium. About 50% of intracellular radioactivity was represented by chromatographically pure cAMP which testifies to the insignificant cAMP metabolism rate. In the experiments with 3T3 and 3T6 cells the influence of possible admixtures and degradation products on 3H-cAMP uptake was revealed. To avoid these influences it is necessary to measure the uptake of cAMP in the presence of theophylline and with abundance of adenosine. In such experimental conditions the intracellular level of cAMP after 1 hour of incubation did not exceed 10% of its extracellular level, which is similar to values obtained on frog muscles. Permeability coefficient of cAMP for membrane of frog muscles and 3T3 and 3T6 cells was about 10(-9)-10(-8) cm X sec-1. PMID- 3027932 TI - A light microscopic and ultrastructural study of two cases of fibrolamellar hepatocellular carcinoma. AB - We describe two cases of fibrolamellar hepatocellular carcinoma of the liver in two young women. Both patients presented with diffuse intra-abdominal metastases; nevertheless they had a survival of 28 and 32 months, respectively, which sustains the better prognosis of this neoplasm. Electron microscopy of one case confirmed the oncocytic features of the neoplastic cells and showed intra- and intercellular duct-like vacuoles with numerous microvilli containing a microfilament core that terminated in a terminal web, which represents an unusual aspect in the spectrum of differentiation of fibrolamellar hepatocellular carcinoma. PMID- 3027933 TI - Reduction of liver glutathione peroxidase activity and deficiency of serum selenium in patients with hepatocellular carcinoma. AB - Twelve adults with hepatocellular carcinoma (HCC) and 8 individuals with histologically normal liver, were measured for serum selenium concentration and glutathione peroxidase (GSH-Px) of liver tissue. It was found a reduced serum selenium and liver GSH-Px in patients with HCC. Serum selenium concentration and the enzyme activity were positively correlated (p less than 0.01). The increased risk of carcinoma in selenium deficiency may be partially due to a reduced activity of GSH-Px, one of the most important scavenger enzymes of oxygen toxic radicals. PMID- 3027934 TI - Successful treatment of malignant trophoblastic diseases in a small oncologic unit. AB - Between 1962 and 1983 a total of 26 patients with malignant trophoblastic disease were diagnosed in northern Finland. The incidence of this disease was 1:21,000 pregnancies. Eight patients had choriocarcinoma and 18 an invasive mole. Clinically, 15 patients belonged to the nonmetastatic and 11 to the metastatic group. Of the latter, 4 patients belonged to the low-and 7 to the high-risk categories. During the first years of the study period, cytotoxic chemotherapy, mostly single-drug therapy, was often complemented with adjunctive surgery and/or irradiation. During recent years, single- or multidrug chemotherapy was supplemented with surgery in only one case with chemotherapy-resistant pulmonary metastases. All 26 patients are alive and disease-free, and after therapy 6 of them have given birth to 1-3 children. Our results suggest that malignant trophoblastic diseases can be successfully treated also in small centers of gynecologic oncology with up-to-date knowledge of the principles of modern cytotoxic chemotherapy. PMID- 3027935 TI - [Deposition of pyrophosphate, chondrocalcinosis and chronic joint disease]. PMID- 3027936 TI - [Leukotrienes in allergic and inflammatory conditions]. PMID- 3027937 TI - [Survival and quality of life in small cell anaplastic pulmonary carcinoma]. PMID- 3027938 TI - Evidence for free radicals produced in aqueous solutions by diagnostic ultrasound. AB - Recent theoretical calculations have shown that small gas nuclei in water exposed to microsecond ultrasonic pulses above an intensity threshold may grow into transient cavities that collapse violently, leading to the formation of .OH radicals and .H atoms. We have detected these free radicals in aqueous solutions exposed to microsecond pulsed ultrasound using spin trapping and electron spin resonance (ESR). The public health implications of our results are discussed. PMID- 3027939 TI - Inhibition of calcium oxalate monohydrate (COM) crystal growth by pyrophosphate, citrate and rat urine. AB - An assay system for the measurement of the rate of Calcium Oxalate Monohydrate (COM) seed crystal growth in a metastable solution of calcium chloride and sodium oxalate containing traces of 14C-oxalic acid was used to assess the inhibitory activity of pyrophosphate (10(-5) M-10(-4) M), citrate (10(-4) M-10(-3) M) and urines of normal and pyridoxine deficient rats. Both pyrophosphate and citrate were strong inhibitors of COM crystal growth and caused a 50% decrease in crystal growth rate at 1.50 X 10(-5) M and 2.85 X 10(-4) M respectively. Normal rat urine strongly inhibited the COM crystal growth, while pyridoxine deficient animals showed a significant (p less than 0.01) decrease in mean inhibitory activity as compared to pair-fed controls. A lowered urinary inhibitory potential accompanied with hyperoxaluria and hypercalciuria, which is known to be associated with pyridoxine deficiency, may be a contributory risk of calcium oxalate crystallization and stone formation. PMID- 3027940 TI - The effects of glycosaminoglycans, pyrophosphate and allopurinol treatment on urease-induced crystallization in vitro. AB - Previous studies have found that human urine inhibits urease-induced crystallization in synthetic urine. To identify the inhibitory components the effects of chondroitin sulphate, heparin and pyrophosphate on urease-induced crystallization were studied and the inhibitory capacity of human urine collected during and before/after allopurinol treatment compared. None of the substances studied nor allopurinol treatment was found to influence the urease-induced crystallization. These findings do not support the idea that glycosaminoglycans or pyrophosphate are responsible for the inhibitory effect detected in human urine. PMID- 3027941 TI - Utilization of cytoplasmic lysozyme immunoreactivity as a histiocytic marker in canine histiocytic disorders. AB - Immunoreactive lysozyme was readily detectable in canine histiocytic disorders including systemic histiocytosis, malignant histiocytosis and granulomatous panniculitis. Lysozyme was less reliable as a histiocytic marker in cutaneous histiocytoma; forty percent of these tumors were negative for lysozyme expression. The marked heterogeneity in lysozyme expression in cutaneous histiocytoma may indicate that a proportion of these tumors show relatively primitive histiocytic differentiation and do not express lysozyme. Alternatively, this same proportion may exhibit a phenotype akin to cutaneous Langerhans cells which do not contain lysozyme. Lysozyme was not detectable in the tumor cells in lymphomatoid granulomatosis, atypical cutaneous histiocytoma, and histiocytic lymphosarcoma. Other evidence that these three disorders do not represent true histiocytic proliferative disorders is discussed. PMID- 3027942 TI - Isolation of parainfluenza-3 virus from bull's semen. PMID- 3027943 TI - Application of ELISA to study avian encephalomyelitis in a flock of turkeys. PMID- 3027944 TI - Papillomas in psittacine birds. PMID- 3027946 TI - Adenovirus associated enterocolitis in a bullock. PMID- 3027945 TI - Horse identification. PMID- 3027947 TI - Duck hepatitis type I and influenza in mallard ducks (Anas platyrhynchos). PMID- 3027949 TI - Turkey rhinotracheitis in France: preliminary investigations on a ciliostatic virus. PMID- 3027948 TI - Preliminary survey for antibodies to bluetongue virus in Indonesian ruminants. PMID- 3027950 TI - Prevalence of bovine virus diarrhoea virus viraemia in cattle in the UK. PMID- 3027951 TI - Isolation of rotavirus from buffalo calves. PMID- 3027952 TI - A comparison of three rapid diagnostic methods for the detection of rotavirus infection in calves. AB - Three techniques for the detection of rotavirus in faecal samples from calves with neonatal gastroenteritis were compared. A preliminary study indicated that reverse passive haemagglutination (RPHA) was at least as sensitive as the enzyme linked immunosorbent assay (ELISA). These two immunoassays were compared with the detection of viral RNA by polyacrylamide gel electrophoresis (PAGE) on 209 field samples. Of the 77 samples in which at least one test gave a positive result, 69 were positive by both RPHA and PAGE, but only 49 were also positive by ELISA, indicating a lower sensitivity for the latter test. The overall agreement between RPHA and PAGE was 96%. The reasons for the discrepancies between the tests are discussed. PMID- 3027953 TI - Characterisation of rotavirus isolates from sub-clinically infected calves by genome profile analysis. AB - Rotaviruses isolated from 43 sub-clinically infected calves from a single farm were analysed by genome profile analysis. The isolates showed genomic variation and eight different profiles were observed, including one which was atypical for Group A rotaviruses. The 3' terminal labelling method for the analysis of genome profiles used in this study required only 1 ng of viral RNA, an increase of 1000 fold in sensitivity over ethidium bromide staining for detecting all rotavirus genome segments. However, dual infections involving two rotaviruses with distinct profiles could not be detected if the concentrations of the viruses differed by greater than 10-fold. PMID- 3027954 TI - Characterization of rotaviruses isolated from pigs with diarrhoea in Venezuela. AB - The prevalence of porcine rotavirus infection was studied in 15 different herds located in the north-western region of Venezuela. The presence of rotavirus was studied by direct electron microscopy (EM) and by an enzyme-linked immunosorbent assay (ELISA). From 136 samples analyzed during the six months of the study (September 1983-February 1984), 38 (27.9%) were found to be positive for rotaviruses, with infection more common in animals that were 4-6 weeks old. Atypical rotaviruses were not detected in any of the samples examined. Most rotavirus positive specimens were subgrouped using specific monoclonal antibodies in an ELISA test. The majority of the samples (26 out of 38) were found to exhibit Subgroup I antigenicity. Only two specimens, collected from the same herd in two consecutive months, were found to belong to Subgroup II. To characterize further the circulating rotaviruses, electrophoretic analysis of the RNA genome was performed on samples selected from nine different herds. Great variability in the RNA electropherotypes was observed. No correlation was found between subgroup specificity and the migration of the two smaller segments (Genes 10 and 11), as has been described for human rotaviruses. PMID- 3027955 TI - Evaluation of an enzyme-linked immunosorbent assay for the detection of transmissible gastroenteritis virus antibodies. AB - An enzyme-linked immunosorbent assay (ELISA) using a detergent-solubilized antigen of purified virus was developed for detection of antibody against porcine transmissible gastroenteritis (TGE) virus in swine serum. The ELISA demonstrated antibody responses in pigs immunized intramuscularly with the attenuated TO-163 strain of TGE virus and in pigs orally infected with the virulent Shizuoka strain of the virus. The results of the ELISA were well correlated with those of the neutralization test. These results indicate the usefulness of the ELISA as a serological tool for TGE virus antibody. PMID- 3027957 TI - [The spread of enzootic bovine leukemia via the seminal fluid in certain breeds of bulls]. AB - Studied was the occurrence of enzootic bovine leukosis as dependent on the use of semen of leukosis-affected bulls for the artificial insemination of cows and heifers and their offsprings in the F1 generation on 16 farms. Semen was used of a total of 30 bulls of the Holstein-Friesian, American Brown, and European Black and-white breeds. The agar gel immuno-diffusion test was employed to establish antibodies to the bovine leukosis virus in the sera of the bulls. On 9 farms with 2,997 cows and heifers that were negative for leukosis antibodies a total of 800 female calves (F1) were born. Serologic investigations of both dams and calves, aged 2 to 5 years revealed no leukosis antibodies. On other 7 farms with 1,717 cows and heifers, among which sporadic carriers of BLV-antibodies were discovered, 713 female offsprings (F1) were born. Seventeen (2.38 per cent) out of these responded positively for BLV antibodies. Twenty-two (3.0 percent) of the dams following calving also showed a positive reaction. Over the 1981-1985 period a total of 1,593 female calves were born as the offsprings of 4,714 cattle on all 16 farms. The percent of the positively responding to leukosis was 1.06, resp., 1.38. These results were considered indicative in ruling out the transmission of enzootic bovine leukosis with semen in the artificial insemination of cows and their F1 offsprings. PMID- 3027956 TI - [Attenuated vaccine against rota- and coronavirus enteritis in calves]. AB - Tested was the effect of the Bulgarian attenuated vaccine of rota- and corona viruses 'PoKo-81', via treatment of dry and freshly calved cows (injected into the udder) and newborn calves (peritoneal injection). It was found that calves fed three times daily on milk obtained from cows that had had intramammary injections did not develop clinical signs of a disease following experimental infection with rota- and corona-viruses. When the cows on infected farms were treated in the same way the disease in calves, offered immune milk, dropped to 36 per cent as against calves of unvaccinated cows, with which morbidity ran as high as 85 per cent. Calves vaccinated via the peritoneum did not fall ill after infection with rota- and corona-viruses on the 6th day following vaccination. In experiments on 18 infected farms morbidity among control, unvaccinated calves was 76 per cent, and mortality--6.8 per cent. With vaccinated calves on the same farms morbidity dropped to 44 per cent, and mortality--to 2.5 per cent. The duration of diarrhea in the vaccinated calves was radically lowered. Discussed is the possibility to enhance the protective effect of the vaccine. PMID- 3027958 TI - [Epizootic enteritis in pigs]. AB - Studied was the etiology of diarrhea in sucking pigs, using animals aged up to seven days, that had suffered from the disease not longer than 24 hours on 12 farms. There were cases of diarrhea induced by the virus of epidemic diarrhea. The virus was characterized morphologically through electron microscopy, and antigenically through direct immunofluorescence and serologic investigation. The disease was distinguished from transmissive gastroenteritis via differential diagnosis. Demonstrated was the lack of cross antigenic relation between the two viruses. PMID- 3027959 TI - [Comparative electrophoretic studies of the virion proteins of virulent and vaccinal strains of Aujeszky's disease virus]. AB - Comparative studies of the virion proteins of a iodo-deoxyuridine resistant, vaccinal strain of the pseudorabic virus--MK-25 and of its parental strain A-2. The analysis in a polyacrylamide gel electrophoresis of polypeptides of highly purified virions of the two strains revealed that they were 18 in number. The electrophoretic mobility of the polypeptides of the vaccinal MK-25 strain was shown to be equal to that of polypeptides of the parental A-2 strain. The molecular weight of 12 of the polypeptides were within the 20,000-70,000 D range. Mostly participating in the two strains were polypeptides of 20,000, 26,000, and 70,000 D. PMID- 3027960 TI - [Development of ELISA for demonstrating antibodies to the swine pestivirus]. AB - An indirect variant of the enzyme linked immunosorbent assay with the classic form of swine fever has been worked out. The test makes it possible to assess the level of antibodies with a great number of animals over a short period of time with no special equipment. The method has proved promising in taking prophylactic measures. PMID- 3027961 TI - [Cytochemical study of acid naphthyl acetate esterase in the lymphocytes of healthy and bovine leukemia virus-infected cows]. AB - A modified cytochemical technique was employed to investigate the unspecific acid naphthylacetate esterase of lymphocytes in the peripheral blood of a total of 64 clinically normal cows, having negative serologic and hematologic reaction for leukosis and of 42 cows with severalfold, well-manifested positive serologic response for the presence of the bovine leukosis virus with no changes in the hematologic data. With the normal animals an average of 6.86 per cent of the peripheral lymphocytes were found to be active--in the form of large dark-brown to dark red-brown granules--and such cells were considered to be T mu mature lymphocytes, while in 53.3 per cent of the cells activity was manifested in the form of tiny powder-like granules, and these were believed to be the Ty lymphocytes. Generally, 63.2 per cent were T-lymphocytes; the remaining 36.9 per cent of the cells had no activity typical for acid esterase, and these were considered to be B-lymphocytes. The study of cows with manifested agar gel immunodiffusion reaction for the presence of the bovine leukosis virus, with no lymphocytosis, revealed that there was a reliable rise of the T mu-lymphocytes up to 23.1 per cent, on an average, of the Ty-lymphocytes up to 69.2 per cent, while the percent of the B-lymphocytes dropped to 7.77 per cent, on an average. PMID- 3027963 TI - Novel nuclear antigens recognized by human sera in lymphocytes latently infected by Epstein-Barr virus. AB - We have used three latently infected cell lines, X50-7, JC-5, and Raji, to identify two new nuclear antigen complexes by Western immunoblotting with human anti-EBNA (Epstein-Barr Virus Nuclear Antigen) sera. One antigen complex, termed EBNA III, is composed of a group of high molecular weight proteins between 130 and 160 kDa and the other antigen complex, termed EBNA IV, is a size-related group of polypeptides between 28 and 62 kDa. Both the EBNA III and EBNA IV groups of proteins display variation in size among the different strains of EBV. Cell fractionation of X50-7, JC-5, Raji, and C16, a cell clone of P3HR1, showed that both new antigen complexes were completely recovered from the nuclei of latently infected lymphocytes as were previously described EBNA I and II. Because these new antigens are only detected by anti-EBNA sera in EBV infected cells, it seems likely that they may be encoded by the viral genome and play some role in the immortalization of lymphocytes by the virus. PMID- 3027964 TI - Comparison of the structural organizations in the 3'-terminal regions of five avian retrovirus strains: RAV 7, RAV 50, B77, PR-B, and SR-B. AB - In order to obtain information on the phylogenies of viral strains which belong to RSV (Rous sarcoma virus) and ALV (avian leukosis virus), the nucleotide sequences of noncoding regions adjacent to the U3 region in two ALV strains, Rous associated virus 7 (RAV 7) and RAV 50, and three RSV strains, Bratislava 77 (B77), Prague:subgroup B (PR-B), and Schmidt-Ruppin:subgroup B (SR-B) were determined by extension from a common primer. The sequences thus deduced were compared with known sequences of other RSV and ALV strains and the structural features of the newly determined viral genomes were discussed. PMID- 3027962 TI - Cloning and nucleotide sequence of Newcastle disease virus hemagglutinin neuraminidase mRNA: identification of a putative sialic acid binding site. PMID- 3027965 TI - The 5' extremity of the v-ets oncogene of avian leukemia virus E26 encodes amino acid sequences not derived from the major c-ets-encoded cellular proteins. AB - Antibodies were prepared against bacterially expressed polypeptides corresponding to various portions of the v-ets-encoded domain of P135gag-myb-ets, the transforming protein of avian leukemia virus E26. Immunoprecipitation analyses show that ca. 80 v-ets-encoded amino-acids located immediately after the v-myb/v ets junction are not found in P54/56c-ets, the translation product of the c-ets proto-oncogene, nor in a set of cellular proteins of 64, 62, and 60 kDa related to but distinct from P54/56c-ets. In addition, Northern blot analyses show that these 5' v-ets sequences neither derive from the nontranslated region of the known cellular transcripts hybridizing to a v-ets probe nor from the c-myb transcript or the helper virus genetic information. Tryptic peptide analyses furthermore indicate that, except for these sequences and the last 16 carboxy terminal amino acids of P135gag-myb-ets, the amino acids encoded by v-ets are essentially colinear with those of P54c-ets. PMID- 3027966 TI - Complete nucleotide sequence of the matrix protein mRNA and three intergenic junctions of human parainfluenza virus type 3. AB - The complete sequence of the gene encoding the matrix protein (M) of human parainfluenza virus type 3 (PIV-3) was determined from cDNA clones and from primer extension dideoxy sequencing of the viral genome. The M mRNA is 1150 nucleotides in length, exclusive of polyadenylate, and codes for a protein of 353 amino acids, having a calculated molecular weight of 39,480. The M protein of PIV 3 was found to have a high degree of sequence homology with that of a closely related paramyxovirus, Sendai virus, and to a lesser extent it contained sequence homology with two more distant paramyxoviruses, measles virus and canine distemper virus. We also determined the sequences of the intergenic junctions for the first four genes of PIV-3: NP, P, M, and F. Comparison of these sequences yielded a consensus mRNA start sequence of 5'-AGGANNAAAGA-3', an mRNA end sequence of 5'-UAAGAAAAA-3', and an intergenic sequence of 5'-CUU-3'. The end sequence of the M gene is unusual in that it contains an eight base insertion prior to the A5 tract found in the consensus sequence. This disruption appears to cause a high frequency of readthrough by the viral transcriptase at this junction. PMID- 3027967 TI - Transcriptionally defective nucleocapsids of vesicular stomatitis virus from cells treated with indomethacin. AB - Indomethacin blocks the biosynthesis of vesicular stomatitis virus (VSV) at the level of primary transcription, RNA replication, and protein synthesis (P. K. Mukherjee and R. W. Simpson (1985), Virology 140, 188-191). Nucleocapsids of infecting virus particles recovered from indomethacin-treated cells were analyzed for in vitro transcriptase activity. Incorporation of [3H]UTP in mixtures containing nucleocapsids from HEp-2 cells pretreated with 10(-3) M indomethacin was inhibited approximately 80% compared to control reactions containing nucleocapsids from untreated infected cells. The level of inhibition of in vitro transcriptase activity of viral nucleocapsids from drug-treated cultures varied according to the cell line used for infection. After indomethacin removal, cells regained their ability to produce enzymatically competent viral-transcribing complexes unless they were subsequently exposed to metabolic inhibitors such as actinomycin D or alpha-amanitin. Enzymatically defective nucleocapsids from indomethacin-treated cells showed enhanced in vitro transcriptase activity in the presence of modulators of prostaglandins and cyclic nucleotides. Electrophoretic analysis of product from in vitro transcriptase reactions revealed that these defective nucleocapsids are unable to synthesize VSV messenger RNA or normal size leader RNA species but only smaller transcripts of undetermined identity. PMID- 3027968 TI - Genome of coxsackievirus B3. AB - The entire nucleotide sequence of the coxsackievirus B3 strain Nancy (CB3) genome has been determined from cDNA. The genome is 7396 nucleotides long, and encodes a 2185 amino acid long polyprotein. It exhibits the same gene organization as other enterovirus genomes. A detailed comparison was carried out between the proteins encoded by the CB3 and poliovirus type 1 strain Mahoney (PV1) genomes. The genes encoding the VPg polypeptide and the viral polymerase are the most conserved regions. The structural polypeptides VP1, VP2, and VP3 are less well conserved although proline and tryptophan residues frequently are found in identical positions. The VP1 protein of CB3 shows a particularly limited homology in those regions which have been found to induce neutralizing antibodies against PV1. The 5' noncoding region of CB3 is closely related to that of PV1, with regard to both length and sequence organization, whereas the 3' noncoding region of CB3 exhibits some unique features. PMID- 3027969 TI - Complete nucleotide sequence of the genome of coxsackievirus B1. AB - The complete nucleotide sequence of the genome of the coxsackievirus B1, a human enterovirus that belongs to the Picornaviridae, was determined by using molecular cloning and rapid sequence analysis techniques. Sequence analysis of the cloned cDNAs revealed that the virion RNA was 7389 nucleotides long and polyadenylylated at the 3' terminus. Similar to other picornavirus genomes, a single large open reading frame was identified. The translated sequence starts at nucleotide position 742 and ends at 7287 of the genome. Thus, the viral polyprotein should consist of 2182 amino acids. When the predicted amino acid sequence of the viral polyprotein was compared with those of other human enteroviruses such as polioviruses, a striking sequence homology was observed, especially in viral proteins 1B, 2C, and 3D. This allowed us to predict precise map locations of the viral structural and nonstructural proteins on the genome, although two proteolytic processing sites, between 1D and 2A and between 2B and 2C, were obscure. The result presented here implied important information with respect to the genetical variation of human enteroviruses. PMID- 3027970 TI - Characterization of the integration site of the CMV mtr in a tumor cell line. AB - Previous studies have shown that a 558-bp fragment of human cytomegalovirus (CMV) DNA contained within pCM4127 and designated CMV mtr can morphologically transform rodents cells in vitro. By cotransformation with pCM4127 and a plasmid conferring G418 resistance, pSV2neo, morphologically transformed NIH3T3 cell lines were isolated. Dot blot hybridization indicated that approximately 30% of the transformants retained CMV sequences. Two cell lines which retained viral DNA were chosen for further study. They were capable of anchorage independent growth and formed tumors in nude mice. Integrated viral sequences in the transformants and tumor cell lines were analyzed by Southern blotting. A bacteriophage lambda library was constructed using a tumor cell line which retained a single copy of the viral sequences, and a phage was isolated which contained the integrated plasmid and the flanking cellular sequences. A complex rearrangement between pCM4127 and pSV2neo had occurred. DNA sequence analysis showed integration of the plasmid sequences into repetitive mouse DNA and identified an adjacent mouse sequence. PMID- 3027971 TI - Protease activation mutants of Sendai virus: sequence analysis of the mRNA of the fusion protein (F) gene and direct identification of the cleavage-activation site. AB - Trypsin cleaves the fusion protein (F) of wild-type Sendai virus into two disulfide-linked polypeptides, F1 and F2, and thereby activates the membrane fusion activity of the virus. A. Scheid and P.W. Choppin [1976). Virology, 265 277) selected mutant viruses of which the F protein could be activated by different proteases, either elastase, chymotrypsin, or plasmin. Herein, we have further characterized five of these mutants. Sequencing of each mutant mRNA encoding the 60-70 amino acids surrounding the cleavage site revealed one or two amino acid changes near or at the cleavage sites. Virions cleaved in vitro by the appropriate proteases were assayed of their fusion activity by hemolysis, and the cleavage sites were determined by amino acid sequencing. In three cases, the change of protease specificity can be accounted for by changed amino acids right at the cleavage site, whereas several other mutations that potentiate cleavage at new sites by new proteases are somewhat removed from the actual cleavage site. We surmise that such mutations might alter local polypeptide conformation, thereby allowing the proteases access to existing sites. Cleavage at new sites produced fusion proteins with novel F1 NH-termini. We found that a mutant with a charged residue at the third position of this normally hydrophobic NH-terminal sequence retains activity in the hemolysis assay, whereas a mutant with a charged residue at the first position does not. PMID- 3027973 TI - The nucleotide sequence of the gene encoding the Newcastle disease virus membrane protein and comparisons of membrane protein sequences. AB - The nucleotide sequence of a cloned cDNA copy of the mRNA encoding the Newcastle disease virus (NDV) membrane (M) protein was determined. A single open reading frame in the sequence encodes a protein of 364 amino acids with a calculated mol wt of 39,742. The predicted protein sequence does not contain extensive hydrophobic regions. The sequence does contain eight pairs of basic amino acid residues, five of which are located in the carboxyl-terminal half of the sequence. Comparisons of the NDV M protein sequence with other paramyxovirus M protein sequences reveals little homology common to all sequences. There is only 17% homology with Sendai virus M protein. A short region of homology with the VSV M protein sequence, was, however, found. PMID- 3027972 TI - Monoclonal antibody study of the subcellular localization and DNA-stimulating activity of murine sarcoma virus-activated transformation-associated proteins. AB - Previously, we reported the monoclonal antibody detection of transformation associated proteins (TAP) in ts110 murine sarcoma virus-transformed normal rat kidney (6M2) cells (Chan et al., 1986). In this study, we used the same monoclonal antibody to investigate the subcellular localization, the fate and the mitogenic activity of TAP, as well as the correlationship between TAP synthesis and the expression of transformation properties of 6M2 cells. It was found that TAP were localized in the cytoplasm (probably the Golgi apparatus) of 6M2 cells. TAP were found as three intracellular polypeptides (mol wt of 66K, 63K, and 60K, respectively), and were rapidly released into extracellular medium. Upon release, TAP changed to two extracellular polypeptides (mol wt of 68K and 64K, respectively). Furthermore, the synthesis of TAP was temperature sensitive and correlated closely with the expression of transformation properties of the 6M2 cells. TAP have been purified by monoclonal antibody-affinity column chromatography and found to have a synergistic effect with insulin in stimulating the DNA synthesis of normal rat kidney cells. PMID- 3027974 TI - A rare common integration site of proviruses of the mouse mammary tumor virus in P-type mammary tumors of mouse strain GR. AB - The mouse mammary tumor virus (MMTV) can induce mammary tumors in mice by proviral activation of the cellular oncogenes int-1 or int-2. Activation of these genes, however, is observed in only a few hormone- and pregnancy-dependent mammary tumors of the mouse strain GR. To study the possible involvement of other oncogenes we cloned three MMTV proviral-host fragments (MT 40, 42, and 53) from different mammary tumors of GR with a single acquired MMTV provirus. From a genomic library of normal mouse DNA we isolated phages with insert DNAs that covered 20-30 kb of the uninterrupted regions. Suitable probes devoid of repetitive DNA sequences were isolated in order to screen other mammary tumors for MMTV proviral integrations in these regions. Only two mammary tumors, MT 40 and 42, showed integration of extra MMTV proviruses within the same region. The integrations occurred only 60 bp apart. The other mammary tumors, however, did not contain MMTV proviral integrations in this region, nor in the MT 53 region. Using mouse-hamster somatic cell hybrid DNA, the MT 40/42 integration region was assigned to mouse chromosome 7, and the second region, MT 53, to chromosome 16. The two regions bear no homology to known cellular oncogenes. We did not observe any mRNA being expressed from these cloned segments either in tumors or in normal mammary glands. These findings indicate that plaque(P)-type mammary tumors in mouse strain GR do not originate from MMTV provirus insertions in a particularly favored integration region, but that there may be a variety of integration sites in these tumors. PMID- 3027975 TI - Poliovirus infection of established human blood cell lines: relationship between the differentiation stage and susceptibility of cell killing. AB - The replication of type 1 poliovirus in 13 established human blood cell lines differing in the differentiation stage and cell lineage was investigated. Three T (CCRF-CEM, CCRF-HSB-2, and Molt-3) and three B (Raji, CCRF-SB, and RPMI 8226) cell lines showed no cytopathic effects (CPE) or virus production. CPE associated with virus production were detected in the other seven cell lines: HL-60, ML-1, and KG-1 (granulocytic lineage), U-937 and THP-1 (monocytic lineage), K-562 (erythroid lineage), and Molt-4 (T cell lineage). These susceptible cell lines greatly differed in the speed at which the CPE progressed. The progression of CPE was faster in relatively well-differentiated cell lines such as HL-60 and U-937, independently of the multiplicity of infection, than in less differentiated cell lines such as K-562, KG-1, and THP-1. Thus, for the same lineage, the speed at which CPE progressed became proportionally higher with subsequent differentiation stages. In the K-562 cell culture, CPE were not observed until at least 5 days postinfection (p.i.), while more than 80% of HL-60 cells were killed within 3 days p.i. There were no significant differences between infected HL-60 and K-562 cells in the efficiency of infection determined at 8 hr p.i. by the indirect immunofluorescent technique, the rate of virus growth, or the amount of viral capsid protein synthesized. This indicated that there were similar viral replication cycles in the two cell lines. These observations suggest that the killing function of the virus is expressed more slowly in K-562 cells than in HL 60 cells. PMID- 3027976 TI - Characterization of a virus-specific proteolytic activity processing the gag precursor of the simian sarcoma-associated virus. AB - The proteolytic processing of the gag precursor polypeptide pr65gag of simian sarcoma-associated virus (SSAV) has been studied in vivo and in vitro. In SSAV infected cells (i.e., in vivo) proteins of 52 and 38 kDa and the viral protein p30 could be immunoprecipitated with anti-p30 serum. This cleavage pattern is only in part imitated by in vitro cleavage of the isolated pr65gag with avian myeloblastosis virus (AMV) protease p15. However, in vitro incubation of isolated pr65gag with detergent-disrupted SSAV particles generated products identical in size to those found in vivo, i.e., proteins of 52 and 38 kDa and p30. The extent of cleavage is dependent on the concentration of the disrupted virions added to the incubation mixture. Studies with protease inhibitors suggest that the SSAV enzyme is a serine-type protease like that of other mammalian retroviruses and unlike the protease of avian viruses. The SSAV protease activity eluted from a molecular sieve column in a range of about 10-15 kDa reflecting the molecular weight of the murine leukemia virus (MuLV) protease (Mr = 13.5K). Thus, it appears that there is a close similarity between the proteolytic enzymes present in different mammalian retroviruses such as MuLV and SSAV. PMID- 3027977 TI - Poliovirion-derived capsid proteins in subviral ribonucleoprotein complexes. AB - Poliovirus containing [35S]methionine-labeled structural proteins was allowed to infect suspension cultures of HeLa cells. The experiments show that a small amount of virion-derived proteins (VDP) attached to the cells and that during infection at 37 degrees C but not at 15 degrees C, these proteins were associated with subviral particles containing RNA. By 3 hr postinfection 53% of the associated VDP was detected in components with low-sedimentation coefficients (below 150 S) and buoyant densities of 1.59, 1.54, and 1.40 g/cc. The rest of the labeled VDP was found mostly associated to a heavy subviral ribonucleoprotein (H vRNP) compatible with a ribosome-RNA complex, due to its ssRNA content, its sedimentation behavior and buoyant density (1.54 g/cc), and its EDTA lability. VDP appeared in this H-vRNP early after infection (60 min) and accumulated as infection proceeded at expense of the low-sedimenting material. The accumulation of VDP in the heavy fraction (HF) paralleled polio-specific protein synthesis, and it was prevented by cycloheximide. These results suggest that a small amount of VDP remains associated with the infecting vRNA, and recommend a model which involves a subviral particle such as a functional molecule probably involved in the initial steps of poliovirus replication. PMID- 3027978 TI - Glycosylation of simian virus 40 T antigen and localization of glycosylated T antigen in the nuclear matrix. AB - Evidence has been obtained for the glycosylation of simian virus 40 (SV40) T antigen in SV40-infected TC7 cells. Both [3H]mannose and [3H]glucosamine are incorporated into T antigen in cells grown and labeled in medium containing fructose instead of glucose. In addition, T antigen is visualized by a carbohydrate stain specific for mannose and/or glucose residues. Finally, lectin binding studies suggest that T antigen contains galactose and/or galactosamine, since T antigen is specifically eluted from soybean lectin by 0.2 M galactose. When gel-purified, [3H]glucosamine-labeled T antigen is subjected to tryptic peptide mapping, label is found in only one peptide, thought to correspond to the methionine-containing peptide extending from Asn-653 to Arg-691, near the carboxy terminal end of T antigen. Insensitivity to tunicamycin and the localization of the glycosylation site in the carboxy-terminus of T antigen, and not at Asn-153, suggest that T antigen is not N-glycosylated. Cell fractionation studies show that [3H]glucosamine-labeled T antigen is preferentially associated with the nuclear matrix of SV40-infected TC7 cells. PMID- 3027979 TI - Characterization of a herpes simplex virus type 2 deoxyuridine triphosphate nucleotidohydrolase and mapping of a gene conferring type specificity for the enzyme. AB - The herpes simplex virus type 2 (HSV-2)-induced deoxyuridine triphosphate nucleotidohydrolase (dUTPase) was purified approximately 600 +/- 43-fold using a combination of affinity, hydrophobic, absorption, and ion-exchange chromatography techniques. The only substrate for the dUTPase was dUTP with a Km of 3.6 +/- 1.1 microM. There was no apparent divalent cation requirement, but the HSV-2-induced dUTPase was inhibited by EDTA (0.1 mM) and this inhibition was reversed by either Co2+ (0.5 mM) or Mg2+ (0.5 mM). The HSV-2-induced dUTPase was distinguished from the HSV-1-induced and cellular dUTPases based upon differences in sensitivity to substrate inhibition, thermostability, and electrophoretic migration in nondenaturing polyacrylamide gels. Analysis of HSV-1 temperature-sensitive (ts) mutants demonstrated that ts A15 and ts K13 did not induce significant amounts of dUTPase activity at the permissive or nonpermissive temperatures. Mutants with defects in HSV-induced DNA polymerase or in the major DNA binding protein induced dUTPase at both temperatures. In contrast ts mutants defective in the alpha polypeptide VP175 (ICP4) did not induce normal levels of dUTPase at the nonpermissive temperature. The location of a gene encoding for the type specificity of the HSV induced dUTPase was mapped to the left 20% of the genome in Us in the region 0.060 to 0.100 or from 0.148 to 0.204. PMID- 3027980 TI - Partial nucleotide sequence of St. Louis encephalitis virus RNA: structural proteins, NS1, ns2a, and ns2b. AB - cDNA clones of the St. Louis encephalitis (SLE) virus genome have been obtained and the nucleotide sequence of 4.7 kb corresponding to the 5' terminal half of the genome determined. The genome contains a 5' noncoding region of 98 nucleotides followed by a single continuous open reading frame that encodes three structural proteins in the order capsid (C), membrane precursor (prM)-membrane (M), and envelope (E). Immediately following the C-terminus of E are located nonstructural proteins NS1 through NS3. The SLE amino acid sequence homology with yellow fever (YF), Murray Valley encephalitis (MVE), West Nile (WN), and dengue-2 (DEN) viruses over the sequenced region is 39, 66, 64, and 43%, respectively. The start of each SLE protein has been assigned on the basis of N-terminal sequence data and potential proteolytic cleavage sites homologous with YF and MVE viruses. Flaviviruses have conserved glycosylation sites in prM and NS1 proteins, although only one of the two glycosylation sites in the SLE E protein is conserved in MVE and DEN viruses. An evolutionary tree showing relationships of SLE, MVE, WN, YF, and DEN-2 flaviviruses is proposed on the basis of the amino acid sequences of the C proteins. PMID- 3027981 TI - The 5'-end sequence of the murine coronavirus genome: implications for multiple fusion sites in leader-primed transcription. AB - The coronavirus leader-primed transcription model proposes that free leader RNA species derived from the 5'-end of the genomic RNA are utilized as a primer for the transcription of subgenomic mRNAs. To elucidate the precise mechanism of leader-priming, we cloned and sequenced the 5'-end of the mouse hepatitis virus genomic RNA. The 5'-terminal sequences are identical to the leader sequences present at the 5'-end of the subgenomic mRNAs. Two possible hairpin loop structures and an AU-rich region around the 3'-end of the leader sequence may provide the termination site for leader RNA synthesis. The comparison of 5'-end genomic sequences and the intergenic start sites for mRNA transcription revealed that there are homologous regions of 7-18 nucleotides at the putative leader/body junction sites. Some intergenic regions contain a mismatching nucleotide within this homologous region. We propose that free leader RNA binds to the intergenic region due to this homology and is cleaved at the mismatching nucleotide before serving as a primer. Thus, the free leader RNA species may be longer than the leader sequences in the subgenomic mRNAs and different mRNAs may have different leader/body junction sites. PMID- 3027982 TI - Multiple recombination sites at the 5'-end of murine coronavirus RNA. AB - Mouse hepatitis virus (MHV), a murine coronavirus, contains a nonsegmented RNA genome. We have previously shown that MHV could undergo RNA-RNA recombination in crosses between temperature-sensitive mutants and wild-type viruses at a very high frequency (S. Makino, J.G. Keck, S.A. Stohlman, and M.M.C. Lai (1986) J. Virol. 57, 729-737). To better define the mechanism of RNA recombination, we have performed additional crosses involving different sets of MHV strains. Three or possibly four classes of recombinants were isolated. Recombinants in the first class, which are similar to the ones previously reported, contain a single crossover in either gene A or B, which are the 5'-most genes. The second class of recombinants contain double crossovers in gene A. The third class of recombinants have crossovers within the leader sequence located at the 5'-end of the genome. The crossover sites of the third class have been located between 35 and 60 nucleotides from the 5'-end of the leader RNA. One of these recombinants has double crossovers within the short region comprising the leader sequences. Finally, we describe one recombinant which may contain a triple crossover. The presence of so many recombination sites within the 5'-end of the genome of murine coronaviruses confirms that RNA recombination is a frequent event during MHV replication and is consistent with our proposed model of "copy-choice" recombination in which RNA replication occurs in a discontinuous and nonprocessive manner. PMID- 3027983 TI - Analysis of intracellular small RNAs of mouse hepatitis virus: evidence for discontinuous transcription. AB - We have previously shown the presence of multiple small leader-containing RNA species in mouse hepatitis virus (MHV)-infected cells. In this paper, we have analyzed the origin, structure, and mechanism of synthesis of these small RNAs. Using cDNA probes specific for leader RNA and genes A, D, and F, we demonstrate that subsets of these small RNAs were derived from the various viral genes. These subsets have discrete and reproducible sizes, varying with the gene from which they are derived. The size of each subset correlates with regions of secondary structure, whose free energy ranges from -1.6 to -77.1 kcal/mol, in each of the mRNAs examined. In addition, identical subsets were detected on the replicative intermediate (RI) RNA, suggesting that they represent functional transcriptional intermediates. The biological significance of these small RNAs is further supported by the detection of leader-containing RNAs of 47, 50, and 57 nucleotides in length, which correspond to the crossover sites in two MHV recombinant viruses. These data, coupled with the high frequency of RNA recombination during MHV infection, suggest that the viral polymerase may pause in or around regions of secondary structure, thereby generating pools of free leader-containing RNA intermediates which can reassociate with the template, acting as primers for the synthesis of full-length or recombinant RNAs. These data suggest that MHV transcription uses a discontinuous and nonprocessive mechanism in which RNA polymerase allows the partial RNA products to be dissociated from the template temporarily during the process of transcription. PMID- 3027984 TI - Fowlpox virus thymidine kinase: nucleotide sequence and relationships to other thymidine kinases. AB - The thymidine kinase (TK) gene of fowlpox virus (FPV) is located in a 2.2-kb HindIII-ClaI fragment derived from a 5.5-kb EcoR1 fragment of the FPV genome. The TK gene was mapped to the region of a 700-bp XbaI fragment contained within this HindIII-ClaI fragment. Nucleotide sequence analysis of this region revealed an open reading frame of 183 codons. Identification of this region as the FPV TK gene was confirmed by its homology with the vaccinia virus TK at both the nucleotide and amino acid levels. The derived FPV TK polypeptide has a calculated molecular weight of 20,380 and is six amino acids larger than the vaccinia virus TK gene product. We have reported previously that the FPV TK gene operates in vaccinia virus without the requirement for a vaccinia virus promoter. The sequence homologies between the two TK promoters substantiated this observation. Northern blot analysis of RNAs from cells infected with a vaccinia virus recombinant expressing the FPV TK gene showed major (700 nucleotide) and minor (1000 nucleotide) transcripts from the FPV TK gene. The deduced amino acid sequence of the FPV TK has significant homology with the TKs from chicken, man, and three other poxviruses, but shows no homology with herpes simplex virus TK. Comparisons of the homologous sequences indicated that the "core" of the enzyme has probably evolved in poxviruses four times as quickly as in vertebrates. Characterization of the FPV TK gene may facilitate the construction of recombinant FPVs as vehicles for the delivery of vaccine antigens to poultry and other avian species. PMID- 3027985 TI - The herpes simplex virus type 1 ribonucleotide reductase is a tight complex of the type alpha 2 beta 2 composed of 40K and 140K proteins, of which the latter shows multiple forms due to proteolysis. AB - We have isolated two monospecific monoclonal mouse antibodies directed against the HSV-1 ribonucleotide reductase. When immobilized to Sepharose, both antibodies remove enzyme activity from solution. However, on immunoblots of crude extracts of HSV-1-infected cells, one antibody only detects a 140K protein and the other antibody only a 40K protein. Neither antibody recognizes the cellular ribonucleotide reductase or the related pseudorabies virus-induced enzyme. Therefore, our data strongly suggest that the HSV-1 ribonucleotide reductase consists of a 140K and a 40K protein. The 140K protein is sequentially degraded to 110K, 93K, and 81K proteins by a Vero cell-specific, N alpha-p-tosyl-L-lysine chloromethyl ketone-sensitive protease. Of the different proteolytic products, at least the 93K species seems to be enzymatically active, suggesting that part of the 140K protein may have functions not related to ribonucleotide reduction. There is a very high affinity between the 140K and 40K proteins as evident from affinity chromatography on antibody-Sepharose and sedimentation velocity centrifugation in a glycerol gradient. The 140K and 40K proteins cosediment with the HSV-1 ribonucleotide reductase activity at 17 S. This indicates that the active form of the HSV-1 reductase consists of the 140K and 40K proteins forming a tight complex of the alpha 2 beta 2 type. PMID- 3027986 TI - The immediate early proteins of varicella-zoster virus. AB - Stable, relatively high-titer varicella-zoster virus (VZV) stocks as well as a high-titer anti-VZV serum prepared in inbred guinea pigs have allowed the identification of VZV immediate early proteins using the classic inhibitor approach. VZV infection was initiated in the presence of cycloheximide. Following the removal of cycloheximide, actinomycin D and radiolabel were added. After the labeling period, extracts were immunoprecipitated with anti-VZV guinea pig serum and subjected to polyacrylamide gel electrophoresis and fluorography or autoradiography. Four immediate early proteins of mol wts 185,000, 69,000, 43,000, and 34,000 were identified. The largest three were phosphoproteins. PMID- 3027987 TI - [Complex x-ray and radionuclide diagnosis of frostbite]. PMID- 3027988 TI - [Disorders of cholinergic regulation and the use of cholinergic blocking agents in bronchial asthma]. PMID- 3027989 TI - [Preoperative radiologic care of patients with juvenile fibroma]. PMID- 3027990 TI - [Adenosine metabolism in lymphocytes and neurochemical stressor reactions in mice with metastatic Lewis carcinoma after surgical removal of the tumor]. AB - Surgical operation of metastatic Lewis carcinoma, carried out in male mice of C57B1 strain, which stimulated distinctly the metastases spreading, was accompanied by phase impairments in activities of adenosine deaminase and 5' nucleotidase in immunocompetent cells correlating with neurochemical stressory reactions. Thus, excessive stressory alterations in activity of symptoadrenal and hypothalamic mediatory systems appear to be among the factors responsible for inhibition of metabolism in lymphoid cells and for stimulation of metastatic spreading. PMID- 3027991 TI - [Changes in lipoxygenase activity in developing granulation tissue during aseptic wound healing in rats]. AB - Activation of lipoxygenases was found during development of granulation tissue in full-layer skin wound simulated in 100 Wistar rats and studied by means of electron paramagnetic resonance and high pressure liquid chromatography. Maximal activation of lipoxygenases within 2 and 6 days corresponded to two phases of wound healing: inflammation phase and phase of fibroblast proliferation during formation of the granulation tissue. PMID- 3027994 TI - [Role of nutrition in acute and long-term therapy of chronic inflammatory bowel diseases]. AB - The clinical picture and course of inflammatory bowel disease are influenced by nutritional abnormalities and malnutrition. Interest at present concentrates on high-fibre low-refined sugar diets, elimination diets with identification of specific food intolerance and low-residue diets. All three failed to show significant positive effects on the course of the disease, need for hospitalisation, surgical procedures required or post-operative recurrence. Only a low lactose diet seems to be justified, since we found lactose intolerance in 25-35% of patients with inflammatory bowel disease, as compared with 5-10% in the normal population. In 25 patients with Crohn's disease (CD) a reduction in inflammatory activity and improvement of nutritional status was obtained with parenteral nutrition (PN). Nevertheless, longer follow up periods revealed no additional benefit in comparison with conventional therapies. Furthermore, the combination of PN and total bowel rest resulted in the same improvement as with PN alone. 25 patients with CD manifesting an acute phase of the condition were treated with tube feeding (TF) as primary therapy. TF reduced CD activity and improved nutritional status in 15 patients with small bowel disease, whereas the patients with colonic disease and extraintestinal manifestations did not react. A comparison of the effect of PN and TF in 10 patients with CD showed no significant difference with regard to clinical course and objective parameters. In view of the high costs and risks of complications of PN, TF is recommended as primary therapy for the acute phase of CD. The importance of substitution therapy, especially of vitamin D, is documented. PMID- 3027992 TI - [Role of the expression of the bovine leukemia virus in the pathogenesis of hemoblastoses]. AB - The role of bovine leukemia virus (BLV) expression was demonstrated in normal, BLV-infected and leukemic cattle. The studies established the changeable character of viremia on the basis of BLV p24 expression in blood plasma and native leukocyte lysates. The experiment revealed a marked regularity: a decrease of antibody titer in blood serum--the appearance of the virus in the body- lymphocytosis. The development of lympholeukemia shortly after experimental infection was detected in animals with an antigen regularly detectable in the plasma. PMID- 3027996 TI - [Psychotherapeutic effects of psychotropic drugs: principle considerations from the psychobiologic viewpoint]. AB - The paper presents, from a psychobiological perspective, a critical and pragmatic discussion on fundamental questions related to the psychotherapeutic effects of psychopharmacological drugs. A series of definitions of the terms "psychopharmacological drugs", "cerebrotropic" and "psychotropic", "psychoactive and psychotherapeutic effects" and psychobiological" are initially given and set within a psychobiological framework of reference. The purpose of this is to reduce inherent contradictions of conventional definitions and to prove the heuristic value of the new concept. The epistemological basis of the presented reasoning is the psycho-physical identity theory that is based on the theories of systems and biological evolution. The author further discusses questions on the neurobiological and psychotherapeutical effects of psychopharmacological drugs, their locus and mechanism of action and other aspects related to the theme. Finally, the problem is raised as to how to separate from each other the multidimensional therapeutic factors, which alltogether result in the final psychotherapeutic effect of the drugs. Among these therapeutic factors, the so called placebo is of prominent importance; the author presents a definition of placebo that fit the conceptual framework of the paper. PMID- 3027995 TI - [Relevance of palpatory, mammographic and thermographic assessment criteria in benign and malignant structural changes in the female breast]. AB - The correlation between clinical findings, mammography and thermography on the one hand and histological structure of the breast on the other hand, was investigated in 133 women. Palpable changes in the breast parenchyma indicated excision biopsy in all cases. The following criteria were employed for palpatory assessment: size, differentiation, nodular mobility and consistency. For mammography, criteria such as calcification, opacity, contralateral comparison and skin thickening were used. For thermography, a modified "Gautherie score" was employed. All three palpatory parameters, however, do not provide reliable differentiation between benign and malignant tumours, since the palpatory findings in fibrocystic disease are similar to those in large malignant growths. Small malignant growths, in turn, may resemble benign tumours. Furthermore, the group of fibrocystic disease markedly reduces the specificity of the radiological findings. However, as aid to differential diagnosis it may be said that apart from the shape of the shadow, contralateral difference in appearance is more likely to occur with benign and malignant tumours than with fibrocystic disease. With regard to thermography a low score i.e. an unconspicuous picture of heat distribution is unlikely to signify a malignant growth. It is, however, impossible to distinguish between mastopathy and malignant tumour. Additional investigations i.e. mammography and thermography do not rule out the existence of a malignant growth. PMID- 3027993 TI - Clinical decision analysis using microcomputers. A case of coexistent hepatocellular carcinoma and abdominal aortic aneurysm. AB - Many difficult medical decisions involve uncertainty. Decision analysis-an explicit, normative and analytic approach to making decisions under uncertainty provides a probabilistic framework for exploring difficult problems in nondeterministic domains. As the methodology has advanced, clinical decision analysis has been applied to increasingly complex medical problems and disseminated widely in the medical literature. Unfortunately, this approach imposes a heavy computational burden on analysts. Microcomputer-based decision support software can ease this burden. PMID- 3027997 TI - [Incidence and significance of cytomegalovirus infections following kidney transplantation]. PMID- 3027998 TI - [Interaction of sex steroids, prostaglandins (6-keto-PGF1 alpha, PGF2 alpha) and beta adrenergic agonists (adrenaline, fenoterol-Partusisten) in the human non pregnant myometrium in vitro]. AB - The in vitro effects of epinephrine and fenoterol (Partusisten) on contractility and prostaglandin (6-keto-PGF1 alpha, PGF2 alpha) production of human, non pregnant myometrial strips dependent on the phase of the menstrual cycle and after pretreatment with estrogens and progesterone are reported. Superfusion of epinephrine (100 ng/ml) stimulated contractility and PG-synthesis of myometrial tissue from the proliferative phase of the cycle, whereas fenoterol (10 ng/ml) had only a little inhibitory effect. After pretreatment with conjugated estrogens (Presomen) the maximal epinephrine-stimulated contractions were correlated to a further increase in PGF2 alpha-synthesis even during fenoterol superfusion. In myometrial tissue obtained during the secretory phase of the cycle, both epinephrine and fenoterol had a significant inhibitory effect on spontaneous contractions associated with an increase in 6-keto-PGF1 alpha-production. After pretreatment with an estrogen-progestin combination (Primosiston) fenoterol superfusion resulted in a complete inhibition of myometrial contractions. These results demonstrate that estrogens increase the myometrial sensitivity to catecholamines leading to increased contractility and increased PGF2 alpha synthesis, whereas progesterone probably mediates the beta-adrenergic inhibitory effect on myometrial contractions. Thus, the contractions inhibiting effect of the betamimetic drug fenoterol associated with decreased PGF2 alpha- und increased prostacyclin (PGI2) synthesis depends on the effect of progesterone in human, non-pregnant myometrium. PMID- 3028000 TI - [Retroviruses and psoriasis vulgaris]. PMID- 3027999 TI - [Effect of pregnancy on metastasizing breast cancer--a case report]. AB - The case of a 39 year old III gravid, I parous woman is reported. Four years ago she had had breast cancer (T2N1MO). Three years later she suffered from recurrence and a tumor was removed from the right axilla. One year later she became pregnant and presented with multiple bone metastases in her 5th month of pregnancy. During the following time an obvious progression could be shown. Therefore, a caesarean section with ovariectomy was performed at the beginning of 29th week of gestation. Postoperatively the patient had a partial remission. It is argued, that this could have been caused by the interaction of pregnancy and malignant disease, especially by the influence of growth-promoting estrogens. PMID- 3028001 TI - Distribution of adenylate kinase (AK) polymorphism in Indian populations. PMID- 3028002 TI - [Maturation of mammalian bone minerals. A new method for determining bone metabolism]. AB - The present report begins by comparing data on bone mineralization communicated in the literature to date with the authors' own previously published concept of the "three-phase model". The age-dependent accumulation of minerals in the femurs of Wistar rats was also chemically-analytically investigated. The results suggest that the mineral first formed is octacalcium phosphate. It is followed by sodium- and magnesium-containing phases. Maturation of octacalcium phosphate only occurs after this; it is converted into a highly carbon-deficient hydroxyapatite. During this conversion the molar Ca/P ratio rises from 1.33 to 2. However, the final value of 2 is not fully attained: this is attributable to the "turnover" activity of bone-metabolic processes. The paper concludes with a discussion of how the newly developed chemical method of determining bone turnover can be used to determine its vitality. PMID- 3028004 TI - [Interrelationship and interaction between the nervous and immune systems]. PMID- 3028003 TI - [Hormonal level and target-cell innervation in the functional development of a beta-adrenoreceptor-adenylate cyclase membrane complex]. PMID- 3028005 TI - [Chance for prolonging life and decreasing the quality of life by intensive chemotherapy]. PMID- 3028006 TI - [Eicosanoids and lung function]. PMID- 3028007 TI - [Approaches to determining vitamin requirements based on an analysis of the interrelation between the levels of actual consumption and the biochemical allowance parameters]. PMID- 3028008 TI - [Hormonal status and emotional characteristics of patients with alopecia areata]. PMID- 3028009 TI - [Pleomorphic adenoma of the salivary glands. Histogenesis, cellular differentiation, tumor marker]. PMID- 3028010 TI - [Inactivation of formaldehyde by semicarbazide in disinfectant tests with viruses]. AB - In suspension tests with viruses formaldehyde is--even in weak concentrations- toxic for tissue cultures. Through addition of semicarbazide in a ratio of one part of 10% semicarbazide to one part of 0.7% formaldehyde, which is equivalent to a molar ratio of 3.8:1, the toxicity will be neutralized if semicarbazide is added to the tissue cultures before their being inoculated with the virus disinfectant-mixture. In qualitative experiments the toxicity is completely neutralized, in quantitative ones it is reduced by a factor of 10 up to 100. Thus the evaluation of the virucidal capacity of disinfectants containing formaldehyde is conspicuously improved. PMID- 3028011 TI - Subdural hematoma caused by cerebral tumors. AB - Three cases of subdural hematoma caused by intracranial tumors are presented. In one case sudden worsening of patient's condition was due to intracerebral and subdural hemorrhage. In the other one tumor manifested by intracranial hemorrhage and the patient was initially diagnosed as SAH. In this case only the capsule of hematoma was found during operation; spontaneous resolution of hematoma took place. In the third case tumor and hematoma were localized in posterior fossa. Two weeks before admission to the hospital the patient develops symptoms of mild intracranial hypertension, probably due to the growth in space occupying lesion volume, which was caused by the origin of hematoma. In all cases tumors were highly vascularized and showed tendency to bleeding. There was no history of trauma in any case. PMID- 3028012 TI - [Multiple adamantinogenic cysts of the alveolar process in cattle, odontodysplasia cystica congenita, in congenital diseases of the jaw and dentition]. PMID- 3028013 TI - A biotin-avidin amplified enzyme immunoassay for detection and quantitation of orthopox virus camel antibodies in dromedaries. PMID- 3028014 TI - [Bovine herpesvirus type 4 (BHV-4): studies on biology and transmission in cattle herds and insemination bulls]. PMID- 3028015 TI - Characterization of a cell culture adapted goat pox virus. PMID- 3028016 TI - [Neurologic disorders in myeloma disease]. AB - A comprehensive examination involving the use of electroencephalography (EEG), rheoencephalography of the main vessels of the head (REG) and vessels of the limbs, electromyography with the application of neurohistological investigation was conducted in 120 myeloma patients. Neurological disorders systematized into syndromes of encephalopathy, local spondylalgia, radiculalgia, myelopathy, meningomyelopolyradicular and encephalomyelopolyradiculoneuropathic syndromes were observed in 91.7% of the cases. The leading role in the pathogenesis of neurological disturbances was played by toxico-dyscirculatory disorders secondary to infiltration of vessel walls by plasmatic cells, dysproteinosis, kidney failure, as well as mechanical impact of the deformed vertebral column on the spinal cord, its radices and vessels. Modern medicated correction of neurological disturbances is considered necessary in myeloma. PMID- 3028017 TI - [Status of muscle tone in rheumatoid arthritis patients]. AB - Muscle tonus in patients with rheumatoid arthritis (RA) was collated with the activity of the pathological process and functional articular insufficiency (FAI) in different variants of polyneuropathy (P). Hetero- or unidirectional hypotension of muscles was the most characteristic sign in RA. In cases of the sensory variant of P, patients with an early period of the disease and 1-0 degree FAI showed severe damage to the central motor neuron, linked with a dissociated type of muscular dystonia (predominantly hypotension of the flexory group of legs) and a peculiar dystonic phenomenon (a vestibular-cerebellar positioning of the wrist). This kind of wrist positioning was directly correlated with the side of the predominant damage to the central motor neuron. In cases of sensory-motor forms of P, patients with degree II-III FAI, in whom lesions to the peripheral motor neuron were predominant, developed unidirectional muscle hypotension. EMG findings served as an adequate reflection of the nature of changes in the muscle tonus. PMID- 3028018 TI - The effect of high protein diet on the regulatory properties of AMP deaminase from chicken liver. AB - Feeding high protein diet for 5 days caused a 3,5-fold and 2-fold increase of the activity of xanthine dehydrogenase (EC 1.2.1.37) and 5-nucleotidase (EC 3.1.3.31) respectively, in chicken liver. Six hours after feeding the high protein diet there was no change in either enzyme activity although a 3-fold increase in the level of serum uric acid was observed. High protein diet considerably decreased the activity of AMP deaminase at low, but not at high substrate concentration. The activity ratio, measured at 10.0 and 0.16 mM AMP increased from 14:1 (low protein diet) to 23:1 and 24:1 after 6 h and 5 days of high protein diet, respectively. It has been suggested that feeding birds a high protein diet may cause transformation of liver AMP deaminase (EC 3.5.4.6) from a low Km form toward a high Km form. PMID- 3028019 TI - [Anterior interosseus nerve syndrome: a case report]. PMID- 3028020 TI - Kinetics of circular DNA molecule digestion by restriction endonuclease. Computation of kinetic constants from time dependence of fragment concentrations. AB - A model for kinetics of circular substrate cleavage by restriction endonuclease was formulated. The aim of the analysis of the model was to extract kinetic constants for all target sites from time-dependence of fragment concentration in reaction products. That was proved to be possible for molecules with an odd number of fragments only. A symmetry of the molecules with an even number of fragment is the cause. A solution for molecules with an odd number of fragments was found and methods for dealing with the other molecules were suggested. PMID- 3028021 TI - Liver-cell adenoma. Case report. AB - A young woman taking oral contraceptives presented with acute, severe abdominal pain. A liver-cell adenoma (weight 1.6 kg), with intrahepatic bleeding and necrosis, was enucleated at elective laparotomy. The clinical presentation of liver-cell adenoma, ranging from incidental finding to sudden death, is reviewed and the evidence linking oral contraceptives to the tumour is discussed. Reported cases of symptomatic liver-cell adenoma have numerically declined in the past decade. Acute surgical treatment is indicated only to control bleeding. In addition to withdrawal of oral contraceptive medication, enucleation of the tumour or hepatic resection is generally recommended as elective, definitive treatment. PMID- 3028022 TI - Ascitic fluid cytologic features of a malignant mixed mesodermal tumor of the ovary. AB - A case of malignant mixed mesodermal tumor of the ovary in a postmenopausal patient presenting with abdominal distension is reported. Cytologic examination of smears of the ascitic fluid showed the presence of adenocarcinomatous and sarcomatous cells (with some of the latter being giant cells) plus numerous unidentifiable cells that bore some resemblance to either mesothelial cells or macrophages. Electron microscopic studies showed a clear differentiation of the adenocarcinomatous and sarcomatous cells from positively identified mesothelial cells and macrophages also present in the ascitic specimen, indicating that the unidentified cells in fact originated in the adenocarcinoma (endometrioid carcinoma), chondrosarcoma and unclassified sarcoma found in the surgical specimen. The differential diagnostic cytomorphologic and electron microscopic features are described in detail. PMID- 3028023 TI - Expression of tumor cell properties in synovial cells in culture. AB - The tumorigenic properties of human rheumatoid arthritis synovial cells in culture were investigated. The synovial cells developed good colonies and secreted plasminogen activator (PA) and collagenase in the cell cultures, as do Hela cells. Since PA and progesterone receptor (PgR) are considered to be end products of estradiol action in breast cancer cells, the estrogen receptor (ER) and PgR content in these cells was also assayed. Large amounts of ER and PgR were detected in the synovial cells in culture, even though these cells are not targets for sex steroids. Study of the cytomorphologic changes in the synovial cells in culture revealed many characteristics generally observed in neoplastic cells. Whether any or all of these observations have any implication in prognosis or therapy in this disease remains to be studied. PMID- 3028024 TI - cAMP-dependent ACTH secretagogues facilitate corticotropin releasing activity of angiotensin II on rat anterior pituitary cells in vitro. AB - Both arginine vasopressin (AVP) and angiotensin II (AII) potentiate the corticotropin-releasing activity of CRF41 via a potentiation of CRF41-induced cAMP production. In perfused rat anterior pituitary cells, AII (10(-8) mol) showed a transitory 2-fold increase in its ACTH-releasing activity, when tested after application extract of rat stalk median eminence. In order to determine whether this facilitating effect on AII corticotropin-releasing activity occurred through cAMP-dependent mechanisms, the ACTH-releasing activity of AII was tested after stimulation with CRF41, AVP or forskolin, three secretagogues with known effects on cAMP production. When given 16 min after CRF41, 10 micrograms/l, AII (10(-8) mol) showed a significant increase (210%) in its ACTH-releasing activity, which returned to the normal level when AII-stimulation was repeated at 32 min (121%) and 48 min (100%). Similarly, forskolin, 3 X 10(-6) mol, produced a significant transitory increase (208%) in the subsequent AII-induced ACTH release whereas AVP, 10 micrograms/l and 100 micrograms/l, had no effect on the following ACTH response to AII. These results suggest that the AII-induced ACTH secretion- which is cAMP independent--nevertheless may be modulated by previously stimulation of the cAMP pathway. PMID- 3028025 TI - Co-purification of bone resorbing activity and adenylate cyclase stimulating activity from human tumours associated with the humoral hypercalcaemia of malignancy. AB - We have partially purified a tumour factor capable of stimulating both bone resorption in vitro and cAMP accumulation in osteoblastic ROS 17/2 cells from three human tumours associated with humoral hypercalcaemia of malignancy. Purification of tumour factor by sequential acid urea extraction, gel filtration and cation-exchange chromatography, reverse-phase high performance liquid chromatography followed by analytical isoelectric focussing provided a basic protein (pI greater than 9.3) with a molecular weight of approximately 13,000 as a major component of the final preparation which retained both the two bioactivities. Bone resorbing activity and cAMP-increasing activity in purified factor correlated with each other. cAMP-increasing activity of the factor was heat- and acid-stable, but sensitive to alkaline ambient pH. Treatment with trypsin destroyed cAMP-increasing activity of the factor. Synthetic parathyroid hormone (PTH) antagonist, human PTH-(3-34) completely inhibited the cAMP increasing activity of the factor. The results suggest that this protein factor, having its effects on both osteoclastic and osteoblastic functions, may be involved in development of enhanced bone resorption in some patients with humoral hypercalcaemia of malignancy. PMID- 3028026 TI - Long-term effects of ACTH combined with angiotensin II on steroidogenesis and adrenal zona glomerulosa morphology in the rat. AB - To test the hypothesis that the trophic action of angiotensin II on the adrenal zona glomerulosa may allow a sustained stimulation of aldosterone by ACTH by preventing the morphological changes of the zona glomerulosa cells into zona fasciculata-like elements we investigated the effects in rats of a 6-day treatment with ACTH (100 micrograms/kg/day) alone or combined with angiotensin II (300 ng/kg/day) on corticosterone and aldosterone production and adrenal morphology. The responsiveness of both steroids to an acute ACTH dose was also studied on the last day of long-term treatment. Morphologic data showed that prolonged ACTH treatment stimulated the growth of zona glomerulosa cells, though it transformed the tubulo-lamellar cristae of mitochondria into a homogeneous population of vesicles. Angiotensin II furthered the trophic effects of ACTH but prevented the mitochondrial transformation. Despite its ability to conserve the well differentiated aspect of the zona glomerulosa cells, the administration of angiotensin II was unable to prevent the fall in the secretion of aldosterone caused by chronic ACTH treatment and its subsequent unresponsiveness to ACTH stimulation. PMID- 3028027 TI - The effect of monocyte-released oxygen metabolites on colony formation of erythroid progenitor cells. PMID- 3028028 TI - Free and conjugated CSF and plasma GABA in Huntington's chorea. AB - Free and conjugated GABA concentrations were measured in CSF and plasma from 28 patients with manifest Huntington's chorea (HC) and 30 age- and sex-matched controls. GABA was determined by ion-exchange chromatography with fluorimetric detection (IE/F). Free and conjugated CSF GABA was significantly decreased in prolonged HC with advanced disease states and was suggested practicable as an additional diagnostic tool. However, in younger patients (less than 40 yrs) with a short period of HC (less than 2 yrs) an overlap with the age-matched normal range indicated GABA measurement inadequate to early diagnosis nor predictive for offspring at risk. An age-dependent decrease of conjugated CSF GABA was observed in patients and controls. The more pronounced decrease in patients might reflect the neurodegenerative feature of HC. PMID- 3028029 TI - Prevention of endometrial abnormalities. AB - Exogenous oestrogens are highly effective in relieving not only the acute symptoms of ovarian failure, such as vasomotor instability and vaginal dryness, but also in conserving postmenopausal bone mass. However, the oestrogen doses needed to achieve these benefits also induce endometrial proliferation. The risk of endometrial hyperplasia and carcinoma are thereby increased and unopposed oestrogen therapy is associated with a high incidence of abnormal vaginal bleeding requiring appropriate, invasive investigations. The cost-effectiveness of therapy and patient compliance are likely to be correspondingly reduced. Various strategies have been proposed to try to overcome the risk of endometrial hyperstimulation and these strategies have been reviewed. Based upon the available evidence, progestogen addition appears the most sensible and has been shown to be effective. It is now clear that progestogens should, in sequential therapies, be administered for 12 days each cycle for maximum protection. Concern has been expressed that the regular withdrawal bleed induced by sequential treatment will reduce patient compliance. Progestogens have, therefore, been added in a continuous fashion to try to prevent endometrial proliferation and thereby induce amenorrhoea. The ideal continuous, oestrogen/progestogen regimen has not yet been developed: all those evaluated to date are associated with a high incidence of breakthrough bleeding which is likely to restrict their use. Progestogens can cause undesirable physical, psychological and metabolic effects. The incidence and severity of side-effects will depend upon the type of progestogen prescribed, the route of administration, and the dose. Minimum effective daily doses of certain types of progestogens have now been established in terms of endometrial protection. Regrettably, few data are available on the physical and psychological effects of these progestogen doses: more information is available on lipid and lipoprotein effects but the data are confused and, at times, contradictory. More research is urgently needed to determine which of these progestogens is most suitable for addition to postmenopausal oestrogen therapy. PMID- 3028030 TI - Hormonal replacement therapy--cardiovascular disease. PMID- 3028031 TI - Perinatal mortality from Rh(D) hemolytic disease in Finland, 1975-1984. AB - Analysis of 41 deaths from Rh(D) hemolytic disease (HDN) in Finland in the past 10 years revealed that 17 occurred in mothers immunized before anti-D IgG was available and three in mothers immunized by blood transfusion in earlier years. Nine deaths were related to loopholes in the anti-D program and could presumably have been prevented by ensuring that all eligible Rh-negative women received anti D IgG after delivery and abortion. Six deaths were due to immunization during a first pregnancy after the 28th week and three to maternal immunization despite anti-D IgG. Immunization from these sources can only be reduced by anti-D IgG injected antenatally as well as postnatally, though the complete eradication of HDN seems to be beyond our grasp. Giving anti-D IgG to Rh-negative women after the birth of a Rh-positive infant or after an abortion has been common practice in Finland, the former since 1969 and the latter since 1971. Although anti-D prophylaxis has been very effective in reducing the incidence of Rh(D) hemolytic disease, new cases continue to occur. Since the prophylaxis fails to prevent perinatal mortality, we decided to discover how the mothers whose infants died from HDN had become immunized in the past 10 years. PMID- 3028032 TI - Angiotensin-converting enzyme in sarcoid and chalazion granulomas of the conjunctiva. AB - Angiotensin-converting enzyme (ACE) was studied immunohistochemically in conjunctival biopsies from 6 patients with systemic sarcoidosis, 4 patients with posterior non-sarcoid uveitis and in specimens from 4 patients with chalazion of the eyelid. Specimens with sarcoid granulomas showed intense ACE-positive immunoreactivity in epitheloid cells of the granuloma, whereas chalazion granulomas did not contain ACE-immunoreactivity. There was no difference in staining patterns between specimens without granulomas. Thus immunohistochemical staining for ACE may be of help in differentiating conjunctival granulomatous tissue of a chalazion from sarcoid granuloma. PMID- 3028033 TI - Timolol inhibits adenylate cyclase activity in the iris-ciliary body and trabecular meshwork of the eye and blocks activation of the enzyme by salbutamol. AB - The enzymatic activity of adenylate cyclase in homogenates and membrane-rich fractions prepared from rabbit iris-ciliary bodies and bovine trabecular meshwork was found to be inhibited by timolol. Treatment of iris-ciliary body homogenates with Triton X-305 resulted in abolition of the inhibitory effect of the drug on the activity of the enzyme. The stimulatory effect of salbutamol on the enzyme was also susceptible to blockade by timolol. It is suggested that the hypotensive action of timolol on intraocular pressure results from structural and functional changes induced on the plasma membranes of the iris-ciliary body and trabecular meshwork by the thiadiazole group of the molecule, and, also, from the occupation of the adrenergic receptors of the iris-ciliary body by the tert-butylamino-2 hydroxypropoxy part of the compound. PMID- 3028034 TI - Neuronal intranuclear inclusion disease. Clinical ophthalmological features and ophthalmic pathology. AB - Monozygotic twin sisters were afflicted by a chronic progressive neurological disease characterized by slurred speech, nystagmus and oculogyral spasms as well as further extrapyramidal and lower motor neuron abnormalities. At autopsy severe loss of nigral and craniospinal motor neurons was noted. In the nuclei of most nerve cell types of the central and peripheral nervous system, roundish inclusion bodies of 3 to 10 microns in diameter were found. Ocular pathology revealed the presence of identical inclusion bodies in the ganglion cells and ganglion cell loss in the posterior retina. Retinal astrocytosis and loss of myelinated axons of the optic nerve were interpreted as reactive features. No inclusions were found in the retinal pigment epithelium. Careful neuro-ophthalmological studies of the first-degree relatives revealed low b-wave of the ERG with other slight aberrations. These were assumed to represent either a carrier or a subclinical state of this presumably inherited disorder. PMID- 3028035 TI - The light reflex on retinal arteries and veins. A theoretical study and a new technique for measuring width and intensity profiles across retinal vessels. PMID- 3028036 TI - The normal fundus fluorescein angiogram and the normal fundus photograph. An analysis of fundus fluorescein angiography, colour and filter photography of the posterior pole of the eye in clinically healthy subjects and in diabetics without obvious lesions ophthalmoscopically. PMID- 3028037 TI - Adrenocortical adenoma with primary aldosteronism in culture. AB - Four adrenocortical adenomata with primary aldosteronism (Conn's adenomata) were examined by light and electron microscopy employing cell culture methods. Unstimulated cells existed as a unit of the clusters, but Conn's adenoma cells reacted to ACTH. The cultured cells produced mostly cortisol under ACTH stimulation as time passed. Lipid droplets in their cytoplasm decreased in number and the smooth endoplasmic reticulum (sER) was well-developed with dilated, anastomosing tubule. Moreover, mitochondria sometimes had tubulovesicular cristae. It is assumed that Conn's adenoma cells are hybrid type cells, which are intrinsically capable of producing cortisol as well as aldosterone. PMID- 3028038 TI - Primary adenoid cystic carcinoma of the esophagus. Report of a case and its histochemical characterization. AB - A case of primary adenoid cystic carcinoma of the esophagus was reported. A 54 year-old male patient had a tumor in the middle third of the esophagus. Barium swallow and endoscopic examination showed a protruding tumor with a shallow ulceration in its vertex. Histological examination of the surgically removed material revealed that it was largely confined to the submucosal layer, with no metastasis to lymph nodes. The tumor cells presented three distinct patterns: globular nest with irregular cystic spaces, trabecular nest, and true tubule. Electron microscopic study revealed no microvilli or triad along cystic spaces in the globular nests. Histochemical study disclosed that cystic spaces and interstitium were rich in glycosaminoglycans. The luminal surface of the true tubules of the tumor, on the other hand, was characteristically lined by sialomucins. The trabecular type-nests differed from the globular nests in showing GSA-II reactivity and containing abundant retinol-binding protein. These results indicate that the three patterns represented different differentiations of the tumor cells. Twenty-three cases of esophageal adenoid cystic carcinoma were found in the literature from 1950 to 1983 and discussed collectively. PMID- 3028039 TI - [Hypotensive responses of different types of Goldblatt hypertensive rat to MK 421]. PMID- 3028040 TI - In vitro study of paraquat effects on malondialdehyde formation in thymus cells. AB - The present experiments have shown that paraquat enhanced both O2- production and oxidation of exogenous NADPH in thymus cells by NADPH-dependent enzyme system. The effects of paraquat on malondialdehyde formation were also NADPH-dependent and opposite in the absence and in the presence of ferrous ions: Paraquat inhibited the NADPH-dependent malondialdehyde formation in the absence of Fe2+; it activated this formation in the presence of Fe2+ and had no effect on the nonenzymic Fe2+- and Fe2+-ascorbate-stimulated lipid peroxidation. It is suggested that the activating effect of paraquat on the enzymic NADPH-dependent MDA formation in the presence of Fe2+ is due to the formation of a powerful oxidant product (FeO2)+ or to some Fe2+-paraquat radical complex, acting similarly to the ADP-chelated ferrous ions. PMID- 3028041 TI - Ontogeny of sympathetic nerves and transport functions in the rabbit choroid plexus. PMID- 3028042 TI - Circulatory events following spontaneous muscle exercise in normotensive and hypertensive rats. AB - In previous studies we have shown that spontaneously hypertensive rats (SHR) develop a running behaviour and, secondary to the running behaviour, develop an endorphin-mediated analgesic effect. In the present study the role of the central endorphin system in the cardiovascular responses to spontaneous exercise in normotensive Wistar Kyoto rats (WKY) and SHR was investigated. The experimental design allowed us to record mean arterial pressure (MAP) and heart rate (HR) continuously for more than 1 week without interfering with the daily activities of the animals. They were active in running wheels during the dark period (19.00 07.00 h) and the activity was accompanied by a marked rise in HR. In SHR, a clear depression of blood pressure lasting for about for about 50 min was noted following each running period. The MAP during the post-running depression was 131.4 +/- 1.6 mmHg which was significantly lower than the pre-running control value (145.2 +/- 2.3 mmHg, P less than 0.01). In contrast, MAP in the post running period in WKY was not significantly different from the pre-running values. In addition, the depression period of SHR had a mean post-running length of 49.7 +/- 3.4 min, which is significantly longer than in the WKYs (37.8 +/- 3.5 min, P less than 0.05). In control rats, naloxone infusion had no effect on blood pressure but a marked bradycardia was observed. In nine running SHR receiving a naloxone infusion, their MAP during the depression period was not different from the control pressure. Our study indicates that endorphin systems are involved in the regulating of blood pressure and HR during muscle exercise in SHR. These systems trigger the transient depression of blood pressure observed immediately after a running period in the SHR. PMID- 3028043 TI - NN'-dicyclohexylcarbodiimide (DCCD) reduces pancreatic NaHCO3 secretion without changing pancreatic tissue ATP levels. AB - To study the mechanism responsible for pancreatic NaHCO3 secretion, the inhibitor NN'-dicyclohexylcarbodiimide (DCCD) was administered to six secretin-infused, anaesthetized pigs. Pancreatic juice was collected from a catheter in the main pancreatic duct. Secretion rate was measured at several arterial pH values in each animal, both before and after DCCD. 15 (14-30) mumol kg-1 body wt DCCD, intra-arterially, reduced pancreatic NaHCO3 secretion from 296 (234-398) to 181 (134-237) mumol min-1 at arterial pH 7.43 (7.42-7.47). Similar fractional reductions of secretion occurred at lower arterial pH. Pancreatic tissue ATP concentration, 1.8 (1.4-2.0) mumol g-1 wet wt, was not changed by DCCD. DCCD, less than or equal to 10(-4) mol l-1, did not change Na,K-ATPase nor carbonic anhydrase activities in separate in vitro assay systems. It is concluded that DCCD reduced pancreatic NaHCO3 secretion by a mechanism not involving ATP depletion nor inhibition of Na,K-ATPase nor carbonic anhydrase activities in pancreatic cells. Because DCCD inhibits proton pumps, DCCD may have reduced NaHCO3 secretion through interfering with a proton pump involved in extruding H+ from HCO-3 secreting cells to interstitial fluid in the pancreas. PMID- 3028044 TI - Neuropeptide Y constricts human skeletal muscle arteries via a nifedipine sensitive mechanism independent of extracellular calcium? PMID- 3028045 TI - [Studies of benzimidazole derivatives. XIII. Synthesis of various 2-alpha-hydroxy and 2-beta-hydroxyalkyl-benzimidazoles]. PMID- 3028046 TI - Tumours of the central nervous system. Proton magnetic resonance relaxation times T1 and T2 and histopathologic correlates. AB - Proton MR relaxation times T1 and T2 were determined in vitro in 136 small specimens of astrocytomas grades I-IV, of oligodendrogliomas, metastases of adenocarcinomas, meningiomas and acoustic neuromas. In addition, 7 samples of peritumoural white matter were analysed. The analysed specimens were studied microscopically in their entirety regarding tumour type and occurrence of necrosis and non-tumour tissue admixture, such as fibrosis and haemorrhage. Most of the gliomas had longer relaxation times than normal white matter and T2 was significantly longer than in the other three tumour groups. The metastases had longer T1 than normal white matter, while T2 varied. The astrocytomas tended to show shorter relaxation times with increasing degree of malignancy, and shortening of T1 and T2 correlating with the proportion of tissue necrosis. Similarly, the metastases with tissue necrosis had shorter T1 and T2 than non necrotic samples. The meningiomas had T1 values comparable with normal cortex, while the T2 values varied. Tumours containing a large proportion of fibrous tissue had shorter relaxation times than the others. Acoustic neuromas had only slightly longer T1 than normal white matter, while T2 was not prolonged. Both T1 and T2 were significantly shorter than in all other tumours studied. Peritumoural white matter had prolonged relaxation times compared with normal white matter, correlating to increased water content. These in vitro differences regarding relaxation times in various types of tumours of the central nervous system, dependent on various types of tissue alterations, should be of interest for the interpretation of in vivo images. PMID- 3028047 TI - Electrophysiological effect of HeNe laser on normal and injured sciatic nerve in the rat. AB - The effect of low energy CW HeNe laser irradiation on normal and dissected nerves in the rat was examined. The methods are described. Results are compared to the laser effect on other living tissues. HeNe irradiation was found to increase significantly the action potentials of the nerves. It was found to be a long lasting effect, keeping an increase in the nerves action potential for more than eight months after irradiation has been stopped. A possible explanation for the way the irradiation acts on the nerve is suggested. PMID- 3028048 TI - Properties of cytosol 5'-nucleotidase and its role in purine nucleotide metabolism. AB - 5'-Nucleotidase which was found first in chicken liver and found to be located in cytosol was purified and characterized. This enzyme is termed cytosol 5' nucleotidase for convenience. Some properties of this enzyme are summarized in Table 7. (Table: see text) The specific activity of cytosol 5'-nucleotidase in chicken liver cytosol is higher than that in rat liver cytosol. In response to a high protein diet the activity of cytosol 5'-nucleotidase in chicken liver increased, concurrently with those of purine nucleoside phosphorylase and xanthine dehydrogenase. Of the three enzymes, the activity of cytosol 5' nucleotidase reached a maximum most rapidly. In rat liver, the activities of these three enzymes did not increase on administration of a high protein diet. From these results the principal physiological function of the cytosol 5' nucleotidase is assumed to be dephosphorylation of IMP as the first step in the pathway of uric acid formation from IMP, which is important in the elimination of nitrogen of amino acids and proteins in a uricotelic animal. An allosteric property of this enzyme is considered to be important for control of adenine and guanine nucleotide pools, especially in connection with the biosynthetic activity of the purine nucleotides in uricotelic animals. PMID- 3028049 TI - Regulation of branched-chain alpha-ketoacid dehydrogenase complex by covalent modification. AB - The branched-chain alpha-ketoacid dehydrogenase complex, like the pyruvate dehydrogenase complex, is an intramitochondrial enzyme subject to regulation by covalent modification. Phosphorylation causes inactivation and dephosphorylation causes activation of both complexes. The branched-chain alpha-ketoacid dehydrogenase kinase, believed distinct from pyruvate dehydrogenase kinase, is an integral component of the branched-chain alpha-ketoacid dehydrogenase complex and is sensitive to inhibition by branched-chain alpha-ketoacids, alpha chloroisocaproate, phenylpyruvate, clofibric acid, octanoate and dichloroacetate. Phosphorylation of branched-chain alpha-ketoacid dehydrogenase occurs at two closely-linked serine residues (sites 1 and 2) of the alpha-subunit of the decarboxylase. HPLC and sequence data suggest homology of the amino acid sequence adjacent to phosphorylation sites 1 and 2 of complexes isolated from several different tissues. Stoichiometry for phosphorylation of all of the complexes studies was about 1 mol P/mol alpha-subunit for 95% inactivation and 1.5 mol P/mol alpha-subunit for maximally phosphorylated complex. Site 1 and site 2 were phosphorylated at similar rates until total phosphorylation exceeded 1 mol P/mol alpha-subunit. The complexes from rabbit kidney, rabbit heart, and rat heart showed 30-40% additional phosphorylation of the alpha-subunit beyond 95% inactivation. Site specificity studies carried out with the kinase partially inhibited with alpha-chloroisocaproate suggest that phosphorylation of site 1 is primarily responsible for regulation of the complex. The capacity of the branched chain alpha-ketoacid dehydrogenase to oxidize pyruvate (Km = 0.8 mM, Vmax = 20% of that of alpha-ketoisovalerate) interferes with the estimation of activity state of the hepatic pyruvate dehydrogenase complex. The disparity between the activity states of the two complexes in most physiologic states contributes to this interference. An inhibitory antibody for branched-chain alpha-ketoacid dehydrogenase can be used to prevent interference with the pyruvate dehydrogenase assay. Almost all of the hepatic branched-chain alpha-ketoacid dehydrogenase in chow-fed rats is active (greater than 90% dephosphorylated). In contrast, almost all of the hepatic enzyme of rats fed a low-protein (8%) diet is inactive (greater than 85% phosphorylated). Fasting of chow-fed rats has no effect on the activity state of hepatic branched-chain alpha-ketoacid dehydrogenase, i.e. greater than 90% of the enzyme remains in the active state. However, fasting of rats maintained on low-protein diets greatly activates the hepatic enzyme.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3028050 TI - Phosphorylation state of HMG CoA reductase affects its catalytic activity and degradation. AB - The expressed catalytic activity of liver microsomal HMG CoA reductase, the limiting enzyme in cholesterol synthesis, is reversibly diminished by phosphorylation in vitro. In intact hepatocytes the expressed activity of HMG CoA reductase is enhanced by incubation of cells with insulin, and diminished by treatment with glucagon or with mevalonate. In the latter situations the level of total reductase activity falls following initial inactivation (phosphorylation) of the enzyme. This observation suggested that the phosphorylated form of HMG CoA reductase is more sensitive to proteolysis. HMG CoA reductase is a 97,000 dalton (97 K) integral protein of the endoplasmic reticulum with a cytosolic domain that includes the catalytic site and serine residues that may be reversibly phosphorylated. In vitro the Ca2+-activated proteolytic enzyme, calpain, generates two catalytically-active fragments: a membrane bound 62 K and a soluble 53 K form of the enzyme which are quantified by specific immunoblot procedures. Cleavage of the native 97 K HMG CoA reductase is enhanced by pretreatment (inactivation) of microsomes with ATP (Mg2+) and liver reductase kinase compared to microsomes pretreated with protein phosphatase. This is reflected in a loss of 97 K reductase and an increase in the soluble 53 K form of the enzyme. Degradation of HMG CoA reductase in hepatocytes is partially blocked by lysosomotropic agents and insulin. A steady state model for the turnover of proteins subject to reversible phosphorylation has been developed which recognizes fractional degradative rate constants for the phosphorylated and dephosphorylated species. PMID- 3028051 TI - Regulation of the activity of hepatic phenylalanine hydroxylase. AB - Rat liver phenylalanine hydroxylase catalyzes the tetrahydropterin-dependent oxidation of phenylalanine to tyrosine, according to equation 1. In addition to the naturally-occurring coenzyme, tetrahydrobiopterin (BH4), certain synthetic analogs of BH4 such as 6-methyltetrahydropterin (6MPH4) have high cofactor activity. (formula; see text) The hydroxylase can be activated by a variety of reversible and irreversible modifications, including those caused by partial proteolysis, by interaction with phospholipids such as lysolecithin, by alkylation of a single sulfhydryl group, by phosphorylation catalyzed by cAMP dependent protein kinase, and by preincubation with its substrate, phenylalanine. All of these modes of activation greatly increase the hydroxylase activity in the presence of BH4, whereas the activity in the presence of 6MPH4 is increased only slightly. The ratio of hydroxylase activity in the presence of BH4 compared to the activity in the presence of 6MPH4, therefore, is a useful index of the state of activation of the enzyme. Of the various activation mechanisms listed above, only phosphorylation of the enzyme and phenylalanine-activation appear to operate in vivo. The evidence indicates that these two regulatory mechanisms act synergistically. Thus, phosphorylation of the enzyme by cAMP-dependent protein kinase is stimulated by phenylalanine, especially in the presence of BH4, (which by itself inhibits), whereas phosphorylation sensitizes the enzyme to activation by phenylalanine. One of the consequences of these interlocking control mechanisms is to enhance the responsiveness of the activity of the hydroxylase to alterations in tissue levels of phenylalanine. As a result, elevated concentrations of phenylalanine can be rapidly metabolized, thereby protecting the fetal and neonatal brain from possible damage by excess phenylalanine. PMID- 3028052 TI - Mechanisms and significance of polyamine stimulation of various protein kinase reactions. AB - An overview of the work on polyamine effects on certain protein kinase reactions is presented. In general, the reactions catalyzed by the messenger-independent protein kinases but not by cyclic nucleotide-, Ca2+-, Ca2+-calmodulin-, and Ca2+ anionic lipid-dependent protein kinases, are markedly enhanced by polyamines. The extent of this stimulation depends critically on the nature of the protein substrate and several other factors. A variety of other polycationic compounds including Co3+(NH3)6, polybrene, and certain aminoglycoside antibiotics exert polyamine-like effects in the same reactions. These observations suggest that the charge properties rather than any strict chemical structure play a role in the action of polyamines. Available data do not support a specific "cofactor" function of these amines for the protein kinases involved in the polyamine stimulable reactions. It appears that the action of polyamines is mediated via their influence on the conformational status of the protein substrates thereby altering the availability of the phosphorylatable sites to the active sites on the protein kinases. Although this notion is supported by several lines of evidence, at present a role of the influence of polyamines on both the substrate and enzyme cannot be ruled out. Possible physiological relevance of the polyamine stimulable protein kinase reactions observed in the in vitro experiments remains problematic in the absence of precise knowledge on the "effective" or free concentrations of intracellular polyamines. PMID- 3028053 TI - DNA-topoisomerases and regulation of cell proliferation. PMID- 3028054 TI - The enzymatic basis of cyclophosphamide specificity. AB - Metabolic activation of cyclophosphamide (CP) by microsomal mixed function hydroxylases yields 4-hydroxycyclophosphamide and aldophosphamide defined as activated CP. Activated CP shows relatively high cancerotoxic selectivity in vivo and cytotoxic specificity in vitro and can be trapped rapidly by reversible reaction of hemiaminal group of the oxazaphosphorine ring with protein thiols to form protein bound activated CP (protein-S-CP). Protein-S-CP stores activated CP in a highly stable form. From pharmacokinetics of activated CP in mice after the injection of cyclophosphamide, it was calculated that about 17% of the CP dose given was stored in a pool of protein bound activated CP lasting for several days. From therapy studies with 4-(S-ethanol)-sulfido-CP in combination with excess of cysteine, it was concluded that the protein-S-CP pool may be that form of activated CP which is mainly responsible for the specific cytotoxic effects in the tumor cells. On the other hand free unbound 4-OH-CP was shown to contribute mainly to overall toxicity. No spontaneous toxicogenation of activated CP was noted under in vivo conditions. 3'-5' Exonucleases were found to hydrolyze 4-OH CP, yielding phosphoramide mustard and acrolein as split products. Because of the low affinity of 4-OH-CP for plain 3'-5' exonucleases, it seems however unlikely that these enzymes play a major role in the antitumor effect of CP in vivo. 3'-5' Exonucleases associated to DNA polymerase like in DNA polymerase delta from rabbit bone marrow or in DNA polymerase I from E. coli are more likely candidates for 4-OH-CP toxicogenation because of the much higher specific activity with 4-OH CP as substrate. In experiments with DNA polymerase I from E. coli, 4-OH-CP was shown to inhibit DNA polymerase activity after toxicogenation by the 3'-5' exonuclease subsite of the enzyme. This suggests an enzyme mechanism based suicide inactivation of the DNA polymerase. Because of the close spatial cooperation of the DNA polymerase and 3'-5' exonuclease subsites with primer/template a site-specific alkylation of DNA is also postulated. Thus we raised the hypothesis that cytotoxic specificity of activated CP is based on the interaction of protein-S-CP (protein bound activated CP) with DNA polymerase/3' 5' exonuclease as the target. In a P 815 mouse mast-cell tumor we determined by means of 5' AMP agarose affinity chromatography two/third of total DNA polymerase to be associated with 3'-5' exonuclease. PMID- 3028055 TI - Regulation of the 2',5'-oligoadenylate system by cyclic adenosine monophosphate dependent phosphorylation. PMID- 3028056 TI - Fructose 2,6-bisphosphate. PMID- 3028057 TI - Release of kallikrein and tonin from the rat submandibular gland. AB - The interaction of neurotransmitters and hormones with specific receptors on the plasma membranes of cells results in enzyme secretion from exocrine glands. However, the effects of agonists on the release of kallikrein and tonin from the rat submandibular gland have not yet been evaluated systematically. The purpose of the present study was to investigate the effects of norepinephrine, isoproterenol, methacholine, and cholecystokinin on the simultaneous release of kallikrein and tonin from the rat submandibular gland. Submandibular gland slices were incubated in vitro at 37 degrees C in a modified Krebs-Ringer medium containing 0.2% each of glucose and bovine serum albumin and bubbled with a gas mixture of 95% O2 and 5% CO2. Glandular kallikrein and tonin secreted into the incubation medium were determined by specific radioimmunoassays. Norepinephrine at 10(-5) M concentration increased kallikrein secretion from a control value of 7.7 +/- 1.5 to 114.7 +/- 26.9 ng/min/mg tissues (p less than .01), and at 10(-4) M concentration kallikrein secretion increased to 265.9 +/- 58.3 ng/min/mg tissue (p less than .01). Similarly, norepinephrine at 10(-5) M enhanced the release of tonin from a basal rate of 4.4 +/- 0.6 to 57 +/- 14.4 ng/min/mg tissue (p less than .05), and at 10(-4) M the rate increased to 91.3 +/- 20.0 ng/min/mg tissue (p less than .01). In contrast, isoproterenol, methacholine, and cholecystokinin did not increase the secretion of kallikrein or tonin. We conclude that the secretion of kallikrein and tonin from rat submandibular glands upon sympathetic stimulation is mediated through stimulation of alpha-adrenoceptors only. PMID- 3028058 TI - Metabolism of bradykinin in isolated perfused rat kidney. Measurement of kininase activity in perfusate and urine. AB - The effect of bradykinin and kininase on renal function and the metabolism of bradykinin in the kidney were examined in isolated perfused rat kidney. In this experimental system, exogenous bradykinin did not affect the water and electrolyte handling by the kidney. Most of bradykinin and kininase in urine were derived from kidney but not from circulating medium. Kininase II may not have a major role in bradykinin destruction by the kidney. PMID- 3028059 TI - Carboxyalkyl peptide inhibitors of kininase II: chiral synthesis. AB - Heretofore, carboxyalkyl peptide inhibitors of kininase II (e.g. N-[1-carboxy-3 phenylpropyl]-Ala-Pro, "enalaprilic acid") have been synthesized by means that yield racemic product. Typically, the secondary amine bond is formed by reacting an amino acid or dipeptide with a 2-keto carboxylic acid ester or imide. The group providing the 2-keto function must be used in excess, and the desired S,S,S isomer must be obtained by resolution procedures. We have developed a procedure whereby enalaprilic acid, RAC-X-64 and related compounds are synthesized stereospecifically and in relatively high yields. PMID- 3028060 TI - Slow tight binding inhibitors of angiotensin converting enzyme. AB - We have found that apparent Ki values of some, but not all, carboxyalkyl dipeptide inhibitors of angiotensin converting enzyme decrease as a function of incubation time. The most potent of the ACE inhibitors tested so far is RAC-X-65 (N-[1(S)-carboxy-3-carboxanilidiopropyl]-L-Ala-L-Pro). When RAC-X-65 is not preincubated with human serum ACE (2.4 X 10(-11) M), the apparent Ki value is 4.4 X 10(-10) M. Preincubation of RAC-X-65 with ACE for 15 min before addition of substrate yields an apparent Ki of 4.1 X 10(-11) M. a 90 min preincubation of the inhibitor with ACE yields an apparent Ki of 1.2 X 10(-11) M, i.e., the reaction of the inhibitor with enzyme is virtually stoichiometric. The enzyme:inhibitor complex is poorly separated by molecular sieve chromatography or by dilution. That such tightly bound complexes are formed in vivo is suggested by the following results: The intravenous ED50 (anesthetized rats) of RAC-X-65 is 9.43 nmol/kg, and the time for half recovery (t1/2) of responsiveness to i.v. angiotensin I, 120 ng/kg, following a cumulative dose of 240 nmol/kg of the inhibitor is 165 min. For comparison, the i.v. ED50 of captopril is 105 nmol/kg, and its t1/2 following a cumulative dose of 240 nmol/kg is 16 min. Implied is the possibility that slow tight binding inhibitors of ACE may be used in a 1 pill per day regimen for the treatment of hypertension. PMID- 3028061 TI - Radiolabelled substrates for angiotensin converting enzyme. AB - Six [3H]benzoyl-tripeptides were prepared and tested as substrates for angiotensin converting enzyme. Each was prepared first as its [4-iodo]-benzoyl analog, and an atom of 3H per molecule was introduced by catalytic dehalogenation in 3H2-gas. Kinetic parameters were measured at 37 degrees C using as buffer 0.05 M Hepes, pH 8.0 containing 0.1 M NaCl and 0.6 M Na2SO4. When the substrates were used at concentrations far below their respective Km values, fractional rates of substrate utilization per unit time for constant enzyme concentration were direct function of respective second order rate constants (Kc/Km). Although absolute values of Kc/Km differed for human enzyme as opposed to rabbit enzyme, relative values of Kc/Km were virtually identical. Similarly, relative rates of substrates utilization during passage through lungs of anesthetized rats were similar to relative values of Kc/Km measured in vitro. Thus, there is now a range of ACE substrates usable, in vitro and in vivo, under conditions of first order enzyme kinetics, conditions under which values of V/Km and Ki can be measured directly. PMID- 3028062 TI - Aggregate anaphylaxis and carboxypeptidase N. AB - Bradykinin (BK) is widely believed to play a role in the pathogenesis of anaphylaxis. To help clarify any such roles, we examined for effects of inhibitors of kininase II (angiotensin converting enzyme, ACE) and "kininase I" (carboxypeptidase N, CPN), on the early course of egg albumin-induced aggregate anaphylaxis in anesthetized guinea pigs. In this model, pulmonary and systemic arterial blood pressure (BP) rise (unless pulmonary fibrillation occurs), lung wgt increases by approximately 60% and pulmonary microvessels are occluded by cell-rich thrombi, all within 5 min of i.v. antigen. The 30 min mortality rate is approximately 2%. ACE inhibitors (BPP9a, Captopril and MK 422; doses up to 140 mumol/kg) do not make anaphylaxis more nor less severe in terms discernible by changes in BP, lung wgt, EKG or intravascular coagulation. In marked contrast, an inhibitor of CPN (2-mercaptomethyl-3-guanidinoethylthiopropionic acid, 2-MGP; 8 16 mumol/kg) increases the 30 min mortality rate to 94% and lung wgt to 180% of control. The animals die in ventricular fibrillation. Given the enormous BK potentiating effects of BPP9a, Captopril and MK 422, it seems likely that little if any BK is formed in the early min of anaphylaxis. 2-MGP does not potentiate BP effects of BK but markedly potentiates effects of C3a anaphylatoxin. Thus, our data support the views that BK is neither a primary nor secondary mediator of aggregate anaphylaxis, and the adverse effects of 2-MGP are best explained in terms of preservation of anaphylatoxins and not in terms of preservation of kinins. PMID- 3028063 TI - Michaelis-Menten kinetics of pulmonary endothelial angiotensin converting enzyme in the conscious rabbit. AB - Estimations of Michaelis-Menten constants Km and Amax (product of Vmax and pulmonary microvascular plasma volume) of pulmonary, endothelial-bound angiotensin converting enzyme (ACE) for the synthetic substrate 3H-Benzoyl-Phe Ala-Pro (BPAP) were performed in rabbits, in vivo, utilizing indicator dilution techniques. The animals were conscious and equipped with permanent right atrial and left carotid catheters. For each determination of Km and Amax, two consecutive bolus injections of BPAP were given into the right atrial catheter, the first containing 0.1 and the second 1622 nmol of substrate, producing first order and mixed order substrate concentrations, respectively, in the pulmonary circulation. Arterial blood was withdrawn at 0.7 sec intervals for 15 sec after each injection and from the family of substrate concentrations thus created (usually 14-20) and resulting range of substrate utilization (10-90%), Km and Amax values were calculated utilizing the Lineweaver-Burk, Eadie-Scatchard, Woolf Augustinsson-Hofstee or Hanes-Woolf transformations of the integrated Henri Michaelis-Menten equation. All four methods produced similar values of Km (10-12 microM), Amax (6-7 mumol/min) and Amax/Km (600-720 ml/min); however, linear regression analysis of data from the Hanes-Woolf transformation resulted in a higher correlation coefficient (0.96 vs 0.86-0.88 for the other three methods). Values for the kinetic constants reported here were similar to those previously reported in anesthetized rabbits utilizing the Woolf-Augustinsson-Hofstee transformation, but higher than those reported for purified rabbit lung ACE, in vitro. PMID- 3028064 TI - In vivo determinations of Ki values for angiotensin converting enzyme inhibitors. AB - We present two methods for calculating Ki values of angiotensin converting enzyme (ACE) inhibitors, such as captopril, in anesthetized or conscious rabbits. Both methods are based on indicator-dilution type determinations of single pass transpulmonary metabolism of the ACE substrate benzoyl-phe-ala-pro (BPAP). The first method involves two determinations of Michaelis-Menten constants Km and Amax (product of Vmax and lung capillary plasma volume) of endothelial-bound ACE for BPAP. Thirty seconds before the second determination of kinetic constants, the inhibitor is administered iv (e.g. captopril, 12 nmol/kg). Comparisons of the apparent Km and Amax values, obtained after the inhibitor to the control values obtained from the first determination, provide Ki values. With the second method, the ratio Amax/Km is obtained, under first-order reaction conditions, before and 30 sec after administration of inhibitor. These apparent and control ratios are used to calculate Ki values. In both methods, plasma levels of the inhibitor at the time of the determination of apparent kinetic constants are estimated by injecting radio-labelled inhibitor (e.g. 3H-captopril), analyzing radioactivity in arterial samples and correcting for plasma protein binding. These methods are potentially applicable to the clinical evaluation of new ACE inhibitors, in vivo, under normal or pathologic conditions. PMID- 3028065 TI - Rat testicular angiotensin I converting enzyme: purification and comparison with rat pulmonary enzyme. AB - Enzymological properties of rat testicular angiotensin I converting enzyme (RT ACE) were compared with those of rat pulmonary angiotensin I converting enzyme (RP-ACE). The molecule of RT-ACE was different from that of RP-ACE with respect to the molecular weight, i.e., the molecular weight of RT-ACE was estimated to be 104 kilo-dalton (kd) and that of RP-ACE (150 kd) on SDS-polyacrylamide gel electrophoresis. On the other hand, the enzymochemical properties of RT-ACE were very similar to those of RP-ACE, with regard to activation by NaCl, optimum pH, Km value for N*-hippuryl-His-Leu-OH hydrolysis and sensitivities to various inhibitors. Therefore, it was speculated that the portions contributing to the appearance of catalytic activity would be similar between RT-ACE and RP-ACE. PMID- 3028066 TI - Kininase II of human seminal fluid: kinetics and inhibition. AB - The activity of kininase II in crude human sperm was measured continuously by measuring the hydrolysis of a blocked tripeptide 3-(2-furylacryloyl)-L- phenylalanyl-glycyl-glycine (1 mmol/l). Mean seminal plasma activity was 335 +/- 61 U/g protein; the Km was 0.7 mmol/l; pH optimum was 8.8 in a 50 mmol/l HEPES buffer and the chloride optimum was 300 mmol/l. This male genital tract enzyme is inhibited by several kininase II inhibitors. Captopril (SQ 14225) showed IC50 = 1.6 X 10(-8) mol/l, with a competitive pattern (Ki = 7.3 X 10(-9)). 3 (Mercaptomethyl)-oxo-piperidineacetic acid showed the same kind of inhibition with an IC50 = 1.8 X 10(-6) mol/l (Ki = 6.8 X 10(-7) mol/l). Enalapril diacid was the most potent inhibitor and had an IC50 of 4.1 X 10(-9) mol/l and showed a mixed competitive and non-competitive inhibition (Ki = 10(-9) mol/Ki' = 9.5 X 10( 10) mol/l). These in vitro inhibition data suggest that, in vivo, such drugs may effect the function of kininase II in the male reproductive system. The observed 50% inhibition constants are comparable to those observed in lung enzyme suggesting similar kinetic properties. PMID- 3028067 TI - Investigations on the functional role of angiotensin converting enzyme (ACE) in human seminal plasma. AB - The investigations were carried out with partially purified angiotensin converting enzyme (E.C.3.4.15.1) from human seminal plasma and from human blood plasma. The Km-constants for angiotensin converting enzyme (ACE) from both sources, estimated by the use of synthetic substrates, were in the same order. The catalytic properties of the enzymes were characterized by a series of known peptidase inhibitors. The male antifertility drug gossypol (1,1',6,6',7,7' hexahydroxy-3,3'-dimethyl-5,5'-bis-isopropyl-(2,2' -naphthalene)--8,8' dicarboxaldehyde) was identified as a potent ACE-inhibitor. The inhibitory constants of several kinins and other biologically active peptides were determined. Any regulatory influence of the peptides investigated on the ACE activity in vivo is not probably. The inhibitor of Zn-containing metalloproteases 2-(N-hydroxycarboxamido)-4-methylpentanoyl-L-alanylglycin e amide) (Zinkov) selectively inhibited ACE from blood plasma, whereas ACE from seminal plasma was not influenced. In seminal plasma the majority of the enzyme is associated with macromolecular structures, identified as membrane vesicles. These vesicles contain also other enzymatic activities usually detectable in seminal plasma. In the male genital tract ACE is synthesized in the prostate, epididymis and testis. As our data indicate ACE seems not to be involved in the regulation of sperm motility. PMID- 3028068 TI - Biological regulation of testicular angiotensin I-converting enzyme. AB - Angiotensin I-converting enzyme (ACE) from rat testes and lung was purified to homogeneity and partially characterized with respect to physicochemical parameters. Additionally, the biological regulation of testicular ACE by gonadotropins and androgens was investigated was investigated and the cell type with which ACE is associated in testes was identified. Rat testicular ACE is a lower molecular weight, isozymic version of the lung enzyme. Partial proteolysis of each isozyme produces different peptide maps, suggesting unique primary structures for each protein. The sensitivity of each isozyme to Co2+, chelators and thermal denaturation is different, a finding that further supports the hypothesis that structural differences exist between the two isozymes. The pituitary gland is essential for the development during puberty and maintenance during adulthood of testicular ACE. In hypophysectomized mature rats, gonadotropins or androgen can maintain ACE activity to near sham-operated levels. ACE activity in testes appears to be associated almost entirely with various stages of germinal cell development. The function(s) of testicular ACE awaits definition. The mechanism of androgen-maintenance of testicular ACE is unclear. Whether androgen specifically induces gene expression of testicular ACE or simply allows for ACE activity to develop in parallel with spermatogenesis is an unresolved question. PMID- 3028069 TI - Role of kininase II (ACE; E.C.3.4.15.1) in the regulation of renin secretion. PMID- 3028070 TI - The effect of alphanapthylthiourea (ANTU)-induced acute injury on lung binding of antibody to angiotensin converting enzyme (ACE). AB - The effects of ANTU-induced acute pulmonary capillary injury on lung and serum ACE functional activity and the specific accumulation of radio-labelled anti-ACE in lung were explored. Rats were injected either with ANTU or the solvent and sacrificed at various intervals up to one week after injection. All ANTU-injected animals developed pulmonary edema and bilateral pleural effusions which resolved by the one week time point. At no time was there any significant change in serum ACE levels. The specific activity of total lung ACE however rose from 11.0 +/- .95 (mean +/- SEM) to 18.4 +/- 1.1 by two hours after ANTU; by 24 hours, however, solubilized lung ACE had fallen significantly to 6.9 +/- .79 (p less than .01). Total lung ACE had returned to control values by one week. In parallel groups of animals the accumulation of 125I-labelled anti-ACE (AA) or normal sheep immunoglobulin (NSG) was compared in control and ANTU-treated rats. The ratio of the radioactivity in the lungs of AA--injected animals to that in NSG--injected animals fell significantly after ANTU administration (5.0 +/- .88 to 1.2 +/- .28 at 2 hours) suggesting that immunoreactive ACE had fallen despite an increase in ACE functional activity. The decreased binding of AA at the early time points perhaps reflects internalization of endothelial cell ACE in response to injury and an inability of the antibody to interact with the enzyme. The reduction in binding at 24 hours (1.38 +/- .47) correlates with a reduction in total lung ACE. ANTI-ACE may be a useful reagent for quantitating endothelial cell damage following lung injury. PMID- 3028071 TI - Isolation and sequencing of an active-site peptide from angiotensin I-converting enzyme. AB - A glutamic acid residue at the active-site of bovine lung angiotensin I converting enzyme was esterified with p-[N,N-bis-(chloroethyl)amino]phenylbutyryl L-[U-14]-Proline (chlorambucyl-L-[U-14C]-L-Proline), an affinity label for this enzyme. The radiolabeled enzyme was digested with BrCN and only 1 of the 30 cleavage peptides resolved by reverse-phase HPLC contained the bound radiolabel. This active-site peptide (Mr approximately 16,000) was digested with trypsin, and the labeled peptide (T-2) was further degraded with thermolysin. The enzyme digest peptides were also resolved by reverse-phase HPLC. Only 1 of the 5 peptides obtained after thermolysin digestion (Th-1, Mr 1290) contained the bound radiolabel. Th-1 (12 residues) was subjected to manual Edman degradation and the following partial sequence was determined: H2N-Phe-Thr-Glu-Leu-Ala-Asp-Ser-Glu. The radiolabel was released at cycle 3 and the amount recovered was equivalent to the amount of PTH-Glu detected on HPLC. Thus, glutamic acid is esterified with chlorambucyl-L-[U-14C]-Proline which confirms our earlier findings. The sequence that we determined is homologous in five residues with the corresponding sequences of carboxypeptidase A and B, two other mammalian zinc-proteases. There is little sequence homology with thermolysin, a bacterial zinc-protease that also contains an essential active-site glutamic acid residue. PMID- 3028072 TI - Value of determination of kininase II in bronchoalveolar lavage fluid. AB - Kininase II (KII), identical with angiotensin-I-converting enzyme (E.C. 3.4.15.1) was characterized biochemically and assayed fluorimetrically in bronchoalveolar lavage fluid and serum of 153 patients with several pulmonary disorders. The albumin concentrations of serum and bronchoalveolar lavage fluid (BLF) have also been measured. The pH optimum of KII derived from BLF (LKII) was 8.0. The Michaelis Menten constant was 38.5 mumol/l using benzyloxycarbonyl-phenylalanyl histidyl-leucine as synthetic substrate. LKII could be inhibited between 80 and 100% by EDTA, phenanthroline, dimercapto-1-propane-sulfonic acid (DMPS), hydroxyquinoline and captopril. The LKII activity (mU/ml BLF) showed no differences in all lung diseases, but the specific LKII (mU/mg albumin) was significantly elevated in sarcoidosis compared to pneumonia (p less than 0.05), fibrosis (p less than 0.05), chronic obstructive bronchitis (p less than 0.005) and lung cancer (p less than 0.01), but not in tuberculosis. This study shows that LKII is measurable in native, unconcentrated BLF and the results indicate that LKII could be useful for diagnosis of pulmonary disorders. PMID- 3028073 TI - Evidence for a potent angiotensin I degrading enzyme different from angiotensin I converting enzyme in rat vascular tissues. AB - There are indications for the existence of an intrinsic renin angiotensin system in vascular walls, which is assumed to participate in blood pressure regulation and in pathogenesis of arterial hypertension. It was evaluated if and to what extent the decapeptide angiotensin (A) I, one of the natural substrates of A I converting enzyme (ACE), is degraded by other peptidases than ACE in rat vascular tissues. A I and A II degradation was studied in arterial and venous vascular wall extracts. The activities ranged between 0.068 +/- 0.025 U and 0.044 +/- 0.025 U. The enzymes involved were biochemically characterized by determination of isoelectric points (pI), pH optima, molecular weights and by investigation of their inhibition behavior in vitro. One potent A I degrading enzyme (AIDE) was identified with pI between 3.6 and 3.9, and pH optimum at 7.75. In vitro studies revealed that AIDE activity was not blocked by the specific ACE inhibitors MK 421 or MK 422 (both 11 nMol/ml). The molecular weight of AIDE ranged between 440,000 and 457,000. The results indicate that AIDE is not identical to ACE (pI 4.2-5.0; pH optimum 8.3). AIDE was also observed in aortic smooth muscle cells cultured in vitro. AIDE decreased following bilateral nephrectomy or administration of aldosterone combined with sodium chloride loading, whereas it was elevated in spontaneously hypertensive rats (Okamoto strain). Since AIDE metabolizes A I, one of the substrates of ACE, it may indirectly affect A II formation and bradykinin inactivation as well. PMID- 3028074 TI - Pharmacological "enkephalinase" inhibition in man. AB - "Enkephalinase", a peptidase capable of degradating enkephalins, has been recently characterized in man, in both plasma and cerebro-spinal fluid (CSF). This study was designed to evaluate the ability of putative "enkephalinase" inhibitors, D-phenylalanine, captopril and thiorphan to decrease "enkephalinase" activity (EKA) in plasma and CSF in human sufferers. All drugs studied decreased plasma EKA. Captopril and thiorphan also decreased CSF EKA. Of the three drugs tested thiorphan proved to be the most potent "enkephalinase" inhibitor in both plasma and CSF. These results show the usefulness of EKA assessment as a procedure for evaluating the potency and specificity of putative "enkephalinase" inhibitors in man. PMID- 3028075 TI - Mechanism and significance of kinin formation in nephrotic syndrome. AB - There were few reports demonstrating behavior of kinin and kininogen in the nephrotic syndrome. In this paper, coagulation factors related to contact activation, such as factor XII (FXII), factor XI (FXI), prekallikrein (PK), high molecular weight kininogen (HMWKG), and kinins were measured in 15 cases of nephrotic syndrome, and clinical significance of these results were discussed. Plasma FXII activity was markedly decreased in the onset and florid stages of nephrotic syndrome, and this decrease was not correlated with plasma albumin level, which suggested marked activation of this factor in these stages. However, the decrease of PK was slight at the above stages. Activations of contact factors were not parallely occurred with the marked consumption of FXII. Plasma kinin activity was not increased in the onset and critical stages of the nephrotic syndrome but increased in the convalescent and chronic stages, while HMWK level was maintained higher than normal throughout the course. Plasma angiotensin converting enzyme (kininase II) activity was increased in the early stage and decreased lower than normal during the course of the disease. It was concluded that kinin formation in the nephrotic syndrome was not due to the activation of intrinsic coagulation system but due to release of kinin from low molecular weight kininogen. This increased level of kinin activity in convalescent and chronic stage may be related to the healing process during the course of this syndrome. Low kinin activity in the early stage of this disease might be also explained by increased kininase II activity. PMID- 3028077 TI - Interrelation between the renin-angiotensin system and kallikrein-kinin system in patients with essential hypertension. AB - In order to investigate the relationship between the kallikrein-kinin (K-K) system and renin-angiotensin (R-A) system, plasma kinin (pKIN), plasma angiotensin II (pAII), plasma renin activity (PRA) and angiotensin converting enzyme activity (ACEA) were determined in 19 essential hypertensives (EHT). pKIN and pATII measurements were performed by highly sensitive radioimmunoassay (RIA), both of which were established in our laboratory. The assay sensitivity of pKIN and pAII were 0.5 pg/tube and 0.1 pg/tube, respectively. In pKIN RIA, pKIN was extracted by ethanol from 0.8 ml of plasma obtained with a syringe containing kinin generating and destroying enzyme inhibitors. In pAII RIA, pAII was measured directly in small amounts of 50-100 microliter, unextracted plasma samples. The level in pAII was significantly higher (p less than 0.05) in the normal renin group (NRH: n = 13) as compared with low renin group of EHT (LRH: n = 6). However, no significant difference was found in pKIN and ACEA between these two groups. Although a significantly positive correlation was observed between pAII and PRA (p less than 0.001) in EHT, the ratio of pAII/PRA tended to be higher in LHR than in NRH. ACEA correlated positively with pAII (p less than 0.01) or PRA (p less than 0.02), respectively. On the other hand, a significant negative correlation was also found between pKIN and pAII (p less than 0.05). From these findings, it was assumed that there was a closed relationship between R-A and K-K systems, and that angiotensin converting enzyme (kininase II) might play some role in the interrelation between both systems. PMID- 3028076 TI - Effects of three different kininase II inhibitors on blood pressure and renal vasoactive substances in essential hypertensives. PMID- 3028078 TI - Comprehensive studies on the renal kallikrein-kinin system in essential hypertension. AB - In order to investigate the role of the renal kallikrein-kinin (K-K) system in normal (NRH) and low renin (LRH) subgroups of essential hypertension (EHT), daily urinary excretions of renal K-K system components including kallikrein (KAL), total KAL, pre-KAL, kinin (KIN) and kininase (total, I and II), were measured in 21 normotensives (NT) and 45 patients with EHT (NRH: 29, LRH: 16). Urinary KAL and KIN quantities, KAL activity, total and pre-KAL, and kininase (total, I and II) were measured by direct RIA, kininogenase assay, direct RIA of KAL after trypsin treatment, and KIN destroying capacity, respectively. The daily excretions of KAL quantity and activity, total and pre-KAL, and KIN were significantly lower in EHT than in NT. That of total kininase and kininase I were significantly higher in EHT than in NT while no significant difference was found in kininase I between EHT and NT. In comparing NRH and LRH, the urinary KAL activity and KIN were lower in LRH than in NRH, and kininase I was higher in LRH than in NRH. No significant difference, however, was found in total and pre-KAL, KAL quantity and kininase II between NRH and LRH. The ratio of KAL quantity/total KAL which reflects the conversion rate from pre-KAL in the kidney, did not show any significant difference among NT, NRH and LRH.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3028079 TI - Studies of the effects of insulin, bradykinin, and captopril on blood glucose levels of alloxan-diabetic rats. AB - Infusion of bradykinin (1 microgram/min) or saline vehicle for 30 minutes into alloxan-diabetic rats produced no change in the very high levels of blood glucose. Furthermore, intravenous injection of captopril (3 mg/Kg body weight) into the diabetic rats did not result in a significant change of glucose concentrations over a period of 60 minutes. However, infusion of bradykinin 15 minutes after intravenous injection of captopril resulted in a marked decrease of glucose levels (p less than 0.01, compared to pretreatment) by 20 minutes after the start of the infusion. Thus, the captopril potentiated the effect of the kinin, possibly by inhibition of kininase II. Using a spectrophotometric assay with Bz-Gly-Gly-Gly as substrate insulin was shown to be an inhibitor of kininase II purified from hog lungs with an I50 of 1.6 X 10(-5)M compared to captopril with I50 of 2.2 X 10(-9)M. Furthermore, it was found that in vivo infusion of as little as 50 mU insulin over a period of 30 minutes, a dose that by itself was ineffective, potentiated the glucose-lowering activity of a bradykinin infusion in alloxanized rats. Interestingly, the infusion of insulin 15 minutes after injection of captopril, at doses of each compound which alone were inactive, did produce a significant fall (p less than 0.005) in glucose concentrations. Overall, the results show that captopril, insulin and bradykinin can interact to promote a reduction in blood glucose of alloxan-diabetic rats. PMID- 3028080 TI - Salivary kallikrein and kininase activities in periodontal diseases. PMID- 3028081 TI - Kininase I and II activities in serum of patients with lung diseases. AB - In serum of 530 patients with various lung diseases and in 70 healthy control subjects, kininase I (E.C. 3.4.17.3) and kininase II (E.C. 3.4.15.1) were measured spectrophotometrically using hippuryl-L-arginine for estimation of kininase Ia (KIa), hippuryl-L-lysine for kininase Ib (KIb) and hippuryl-L histidyl-L-leucine for kininase II (KII). KIa and KIb were significantly elevated (p less than 0.02) in lung cancer and sarcoidosis, compared to tuberculosis and healthy controls. There was an increase (p less than 0.05) in lung cancer in relation to sarcoidosis, chronic obstructive bronchitis, tuberculosis, pulmonary fibrosis and healthy control subjects. KII was significantly elevated in sarcoidosis (p less than 0.0001). According to the histological types of lung cancer, no differences of KIa, KIb and KII have been found. The ratio KIa/KIb X KII was 2.3 in lung cancer and 6.7 in the group with sarcoidosis. These results show that the determination of kininases can be used for diagnosis of lung diseases. PMID- 3028083 TI - Forskolin: its biological and chemical properties. PMID- 3028082 TI - Effect of kallikrein-kinin system on calcium-vitamin D3 metabolism. AB - Kallikrein has been reported to stimulate callus formation and cell proliferation. It is well-known that vitamin D3 play an role on the metabolism of calcium. However, the relation between vitamin D3 and kallikrein are still poorly understood. We have studied the effect of kallikrein on calcium and vitamin D3 with canine kidney. Mongrel dogs were anesthetized with sodium pentobarbital and prepared by the method of Nakanishi et al. Drugs were infused into renal artery for 60 minutes under three different serum calcium concentrations (Exp. I, Exp. II, Exp. III). Blood samples were collected at the midpoint of the 60 minutes urine collection period. Plasma vitamin D3 concentrations were measured by the metabolites of 25-hydroxyvitamin D3 and 1 alpha,25-dihydroxyvitamin D3. 25 hydroxyvitamin D3 was estimated by competitive protein binding assay and 1 alpha,25-dihydroxyvitamin D3 was determined by radioreceptor assay after separated using HPLC. Electrolytes of sodium, potassium, chloride, calcium and phosphorus in serum and urine, plasma cyclic AMP, glomerular filtration rate, renal blood flow and blood pressure were also measured respectively. Plasma 1 alpha,25-dihydroxyvitamin D3 concentration was found to be decreased by the infusion of kallikrein (0.02 KU/kg/min) in three experimental conditions, especially in Exp. II. Bradykinin (0.02 microgram/kg/min) also caused to kallikrein-like changes of vitamin D3. It is suggested that kallikrein-kinin system is related to inhibit the mechanism of vitamin D3 activation system on kidney. PMID- 3028084 TI - Desensitization of hormonal stimuli coupled to regulation of cyclic AMP levels. PMID- 3028085 TI - Mechanisms involved in calcium-mobilizing agonist responses. AB - Many hormones and neurotransmitters exert their biological effects by increasing the levels of Ca2+ and 1,2-diacylglycerol in their target cells. Major agonists that act in this way are epinephrine and norepinephrine, acetylcholine, vasopressin, cholecystokinin, and angiotensin II. These and other Ca2+-mobilizing agonists may also produce effects that are not mediated by Ca2+ or diacylglycerol, but involve separate receptors and an increase or decrease in cyclic AMP. The general mechanisms by which Ca2+-mobilizing agonists induce their physiological responses are depicted in Fig. 12. These responses appear to involve an initial mobilization of Ca2+ from endoplasmic reticulum and perhaps other intracellular Ca2+ stores, followed by alterations in the flux of Ca2+ across the plasma membrane. The Ca2+ changes are consistently associated with increased turnover of cellular phosphoinositides. The most rapid response is breakdown of phosphatidylinositol 4,5-P2 in the plasma membrane, and there is much evidence that this involves a guanine-nucleotide-binding regulatory protein similar to those involved in the regulation of adenylate cyclase. Myo-inositol 1,4,5-P3 produced by phosphatidylinositol 4,5-P2 breakdown rapidly releases Ca2+ from endoplasmic reticulum, and it is likely that it is the long-sought second message for the Ca2+-dependent hormones. 1,2-Diacylglycerol, the other product of phosphatidylinositol 4,5-P2 breakdown, also acts as a second message in that it activates protein kinase C, a Ca2+-phospholipid-dependent protein kinase, by lowering its requirement for Ca2+. The cellular substrates for protein kinase C and its role in the different physiological responses to the Ca2+-mediated agonists are currently being defined. The major intracellular target for Ca2+ is the Ca2+-dependent regulatory protein calmodulin. This binds Ca2+ with high affinity, and the resulting complex interacts with a variety of enzymes and other cellular proteins, modifying their activities. A major target is the multifunctional calmodulin-dependent protein kinase that phosphorylates and alters the activities of many proteins, for example, glycogen synthase and tyrosine hydroxylase. Calcium ions may also stimulate calmodulin-dependent protein kinases that are more specific, such as phosphorylase kinase and myosin light-chain kinase. Other important Ca2+-calmodulin targets are the microtubule associated proteins, but it is likely that many more will be found.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3028086 TI - Choline plasmalogens, glycerophospholipid methylation, and receptor-mediated activation of adenylate cyclase. PMID- 3028087 TI - Early events in insulin actions. PMID- 3028088 TI - Cyclic AMP and learning in Drosophila. PMID- 3028089 TI - Antireceptor or antihormone autoimmunity and its relationship with the idiotype network. PMID- 3028090 TI - Erythropoietin: origin, bone marrow receptors, and relation to renin substrate (angiotensinogen). PMID- 3028091 TI - Leukotrienes and prostaglandins in the immune system. PMID- 3028092 TI - Modulation of eicosanoid production due to mast cell-macrophage interaction. AB - The experiments discussed above demonstrate that macrophages and possibly PMN modulate and contribute to immediate hypersensitivity reactions. Antigen stimulation of sensitized mast cells in vitro in the presence of macrophages showed that the latter contribute prostaglandins and modulate the 5-lipoxygenase products. Early in the time course, 5-lipoxygenase products, including leukotrienes, were enhanced and later decreased. The decrease appeared to be due to active oxygen species and may serve as a negative feedback mechanism to maintain relative homeostasis. PMID- 3028093 TI - Activation of the superoxide forming NADPH oxidase in a macrophage-derived cell free system by fatty acids and detergents. PMID- 3028094 TI - Myeloid differentiated HL-60 cells display specific leukotriene B4 binding sites. AB - Exposure of HL-60 cells for 6 days to a combination of 1.25% (v/v) dimethyl sulfoxide (DMSO) and 10 microM dexamethasone (DEX) induces myeloid differentiation which results in a cell with many of the characteristics of a mature granulocyte. At 4 degrees C myeloid differentiated, but not undifferentiated, monocytic differentiated or eosinophilic differentiated HL-60 cells display marked specific leukotriene B4 binding. Leukotriene B4 binding at 4 degrees C reaches a maximum within 10 min, is readily reversed by unlabelled leukotriene B4, and is stereospecific. Only molecules with structural and biological similarity to leukotriene B4 can competitively inhibit leukotriene B4 binding. Scatchard analysis at 4 degrees C in differentiated cells shows two classes of binding sites. The high affinity sites have a Kd of 0.27 nM and a Bmax of 14.8 fmoles/10(7) cells; the low affinity sites have a Kd of 0.58 microM and a Bmax of 2453 fmoles/10(7) cells. The appearance of specific leukotriene B4 binding sites in the myeloid differentiated cells correlates with their ability to chemotax in response to leukotriene B4. Undifferentiated cells do not chemotax to leukotriene B4. At 37 degrees C leukotriene B4 is incorporated into phospholipid and triglyceride species in both undifferentiated and myeloid differentiated HL-60 cells making binding studies at 37 degrees C in intact cells impossible. Interestingly, incubation of the cells with cyclooxygenase inhibitors during differentiation enhances receptor density. No evidence of omega hydroxylase activity was found in HL-60 cells. These data suggest that the HL-60 cell may be an excellent model system for the study of leukotriene B4 receptor binding, processing and gene expression. PMID- 3028095 TI - Inositol phospholipids and GTP-binding proteins in signal transduction in stimulated human platelets. PMID- 3028096 TI - Phosphoinositide turnover, Ca2+ mobilization, protein kinase C activation and leukotriene action in pituitary signal transduction: effect of gonadotropin releasing hormone. PMID- 3028097 TI - The role of calcium in leukotriene production: immune response induces cardiac myolysis. PMID- 3028098 TI - Regulation of leukotriene biosynthesis by N-3 fatty acids, cytokines, and stimuli for cell activation. PMID- 3028099 TI - Leukotrienes and prostanoids in health and disease. PMID- 3028100 TI - L-649,923, sodium (beta S*, gamma R*-4-(3-(4-acetyl-3-hydroxy-2 propylphenoxy)propylthio)-gamma-hydroxy -beta-methylbenzenebutanoate. A selective, orally active leukotriene D4 receptor antagonist. PMID- 3028101 TI - Stimulus-response coupling mechanisms in human platelets. PMID- 3028102 TI - On the role of guanine nucleotide binding regulatory proteins (G-proteins) in signal transduction in human platelets: studies with sodium fluoride (NaF). PMID- 3028103 TI - On the regulatory role of the cytoskeleton in the expression of platelet procoagulant activity. PMID- 3028104 TI - Inhibition of human platelet aggregation by motapizone via an increase in intracellular cAMP. PMID- 3028105 TI - Platelet membrane glycoproteins and their role in platelet adhesion and aggregation. PMID- 3028106 TI - Receptors and receptor mechanisms of endogenous platelet stimulatory and inhibitory mediators. PMID- 3028107 TI - Thrombin receptor antagonists. PMID- 3028108 TI - Changes in cytosolic free calcium induced by platelet-activating factor in rabbit platelets: specific inhibition by BN 52021 and structurally related compounds. AB - Increases in cytosolic free Ca2+ concentration induced by PAF-acether in rabbit washed platelets were recorded by using the fluorescent Ca2+ indicator Quin 2. In the presence of 1 mM external Ca2+, PAF-acether 2 X 10(-9) M has been found to increase the intracellular level of free calcium from a basal level of 135.0 +/- 26.9 nM to 2.0 +/- 0.7 microM. Pretreatment of platelets with the PAF-acether receptor antagonists BN 52020, BN 52021 or BN 52022 inhibited dose-dependently the PAF-acether-induced fluorescence signal. This activity was specific for PAF acether, as shown with BN 52021, the most active derivative, which at 3 X 10(-6) M totally abolished PAF-acether effect without modifying thrombin - or calcium ionophore-induced signal. Kadsurenone, another PAF-acether receptor antagonist exhibited similar efficiency as BN 52021 against PAF-acether stimulation. Studied on PAF-induced aggregation of washed rabbit platelets: BN 52020, BN 52021 and BN 52022 exhibited the same potencies for inhibition as on the Quin 2 signal induced by PAF-acether; their activities were BN 52021 greater than BN 52020 greater than BN 52022. The results confirm that these derivatives, which are structurally closely related act at receptor level. The difference in their efficiencies seem to prove that the binding on its receptor site needs a highly defined chemical structure. They could be useful tools for structure-activity relationship studies in order to elucidate the conformation of the PAF-acether binding site. PMID- 3028110 TI - Efficacy of thyroid scintigraphy in the diagnosis of intrathoracic goiter. AB - For evaluation of the usefulness of thyroid scintigraphy in the diagnosis of intrathoracic goiter, we analyzed the results of radionuclide thyroid scintigraphy in 54 consecutive cases with suspected upper mediastinal masses. Intrathoracic goiters were found in 42. The sensitivity, specificity, and accuracy of the scintigraphy for intrathoracic goiter were 93% (39/42), 100% (12/12), and 94% (51/54), respectively. Scintigraphic morphology, scanning technique, and pitfalls are discussed. The results show that most intrathoracic goiters do have thyroid function and that radioiodine scintigraphy is a definitive and cost-effective diagnostic procedure for this disease. PMID- 3028109 TI - [Testicular scanning in intrascrotal lesions]. AB - Testicular scanning with technetium-99m sodium pertechnetate was performed on 152 patients with a variety of intrascrotal lesions. Scrotal images were obtained serially in the perfusion, tissue phase, illustrating the features of each phase in various clinical conditions. The relationship of scrotal imaging to the overall clinical presentation and evaluation of these patients is emphasized in testicular torsion of the testicular appendix, epididymitis, abscess, trauma, tumor, spermatocele, and varicocele. Technical problems of the scanning are also discussed. PMID- 3028111 TI - Hepatic embolization through periportal collaterals: balloon occlusion technique. PMID- 3028113 TI - Clinical significance of early myocardial 99mTc-pyrophosphate uptake in patients with acute myocardial infarction. AB - Early 99mTc-pyrophosphate (PYP) scintigrams of 29 patients receiving coronary thrombolysis induced by urokinase, 3.9 +/- 1.2 hours after the onset of acute myocardial infarction (AMI), were evaluated. Intravenous 99mTc-PYP scintigraphy was performed 2.8 to 8.0 hours after the onset of AMI. All 19 patients with positive findings on early scintigrams had good recanalization, compared to only 3 of the 10 patients with negative findings. The seven patients with unsuccessful recanalization after coronary thrombolysis had total occlusion or subtotal occlusion with delayed washout of contrast material, and three of them had collaterals. The sensitivity, specificity, and predictive accuracy of positive results of early scintigraphy in predicting the presence of early reperfusion were 73%, 100%, and 90%, respectively. Patients with positive early scintigrams showed increasing regional ejection fraction and decreasing thallium defect scores from the acute stage to the chronic stage. These findings indicate that early 99mTc-PYP scintigraphy is a sensitive noninvasive technique for detecting early recanalization in infarcted vessels. The collateral flow or antegrade flow with delay is not enough to cause 99mTc-PYP uptake in a very early stage of AMI. PMID- 3028112 TI - MR imaging: possibility of tissue characterization of brain tumors using T1 and T2 values. AB - To evaluate the usefulness of T1 and T2 values for tissue characterization of brain tumors, 37 histologically confirmed brain tumors were examined with a 0.5-T superconductive MR system. With spin-echo and inversion-recovery imaging sequences, computed T1 and T2 images were reconstructed, and T1 and T2 values of the tumors were calculated. Relaxation rates (1/T1 and 1/T2), T1/T2 ratios, and malignancy indexes, which were originally designed for gastrointestinal tumors, were also calculated. Values of all these parameters were so wide-ranged that it was impossible to characterize the tumor tissue types. PMID- 3028114 TI - The interaction of high-density lipoproteins with extrahepatic cells. PMID- 3028115 TI - Eicosapentaenoic acid: its relevance in atherosclerosis and coronary artery disease. PMID- 3028116 TI - Calcium entry blockers for systemic hypertension. AB - Calcium entry blockers appear to be effective antihypertensive agents in both young and older patients. Studies comparing diltiazem and placebo, diltiazem and propranolol, and diltiazem and hydrochlorothiazide indicate that this calcium entry blocker is more effective than placebo, equally effective as the beta adrenergic inhibitors at least in short-term studies, but not as effective in lowering systolic blood pressure (BP) when compared with hydrochlorothiazide (diastolic BP -11.5 mm Hg with diltiazem vs -12.2 mm Hg with hydrochlorothiazide; systolic BP -12.2 mm Hg with diltiazem vs -20.0 mm Hg with hydrochlorothiazide). Combination therapy with diltiazem and hydrochlorothiazide proved effective in a high percentage of mono-therapy nonresponders. The most common adverse reactions to diltiazem included headaches, dizziness and edema. The exact place of calcium entry blockers in therapy as initial or step-2 therapy with a diuretic in hypertension must be determined by additional long-term experience in large numbers of patients. PMID- 3028117 TI - Implications of local angiotensin production in cardiovascular physiology and pharmacology. AB - The traditional concept of the renin-angiotensin system is a circulation-borne endocrine system whose components are secreted by different organs, i.e., renin from the kidney, angiotensinogen from the liver and angiotensin-converting enzyme from the lung. The product of the biochemical cascade, angiotensin II, acts on specific receptors on multiple target organs. Recent data, however, demonstrate that renin and angiotensin are synthesized locally in many tissues. The emerging concept--that angiotensin is produced locally at tissue sites by an endogenous renin-angiotensin system--has important implications to our understanding of cardiovascular homeostasis. This concept implies that local angiotensin concentrations may exceed those of plasma levels and may play an important role in the tonic control of vascular resistance, cardiac and adrenal functions as well as local intrarenal events. An important mechanism of action of converting enzyme inhibitors may be the blockade of tissue angiotensin generation. Hence, the tissue distribution and kinetics of converting enzyme inhibitors may be an important determinant of drug action. These findings have led us to speculate that aberrant tissue renin, angiotensinogen gene expression(s) or abnormalities in the regulation of the local renin-angiotensin system may result in such cardiovascular disorders as vasospasm, hypertension and cardiac hypertrophy. PMID- 3028118 TI - Effects of calcium on vascular smooth muscle contraction. AB - Calcium initiates smooth muscle contraction by binding to calmodulin and activating the enzyme myosin light chain kinase. The activated form of myosin light chain kinase phosphorylates myosin on the 20,000-dalton light chain and contractile activity ensues. Calcium may also enhance smooth muscle contractile activity by binding directly to myosin, the main component of the thick filament. Recent studies raise the possibility that the calcium-calmodulin complex may also modulate smooth muscle contractile activity by removing the inhibition imposed by caldesmon, a protein that is bound to the thin (i.e., actin-containing) filaments of smooth muscle. In vitro studies have demonstrated that the calcium-activated, phospholipid-dependent kinase, protein kinase C, can phosphorylate smooth muscle myosin at a different site than does myosin light chain kinase and down-regulate its actin-activated magnesium adenosine triphosphatase activity. This raises the possibility that protein kinase C phosphorylation of myosin may play a role in modulating vascular contractile activity in vivo. PMID- 3028119 TI - Evaluation of nuclear magnetic resonance spectroscopy for determination of deuterium abundance in body fluids: application to measurement of total-body water in human infants. AB - Nuclear magnetic resonance (NMR) spectroscopy was used to quantitate abundance of 2H in body water of human infants. This method provides precise measurement of total-body water without the extensive sample preparation requirements of previously described methods for determination of 2H content in body fluids. 2H2O (1 g/kg body weight) was administered to infants and saliva and urine were collected for up to 5 h. An internal standard was added directly to the fluid specimen and 2H enrichment in water was measured by NMR spectroscopy. Working range of deuterium abundance was 0.04-0.32 atom %. Coefficients of variation for saliva samples at 0.20 atom % 2H was 1.97%. 2H content in urine and saliva water reached a plateau by 4 h after administration, and amounts in the two fluids were virtually identical. Mean total-body water determination for six infants was 58.3 +/- 5.8% of body weight (range 53-66%). PMID- 3028120 TI - Prognostic factors in primary mucinous breast carcinoma. AB - Two hundred seven mucinous breast carcinomas were morphologically classified into two different groups: pure mucinous carcinomas consisting only of areas with small epithelial islands of solid tumor floating in abundant extracellular mucin, and mixed carcinomas where the tumor contains large areas of mucin, as well as areas with infiltrating carcinoma devoid of extracellular mucin. Of the 207 tumors, 95 were of the pure type and 112 of the mixed type. The pure and mixed carcinomas differed significantly with respect to a number of prognostic factors, the most important of which was lymph node status. Patients with pure mucinous carcinomas had significantly fewer lymph node metastases at the time of primary operation (P = 0.0001) and a significantly longer recurrence-free survival (P = 0.03) than patients with mixed carcinomas. Patients with mixed carcinomas closely resembled patients with infiltrating ductal carcinomas not otherwise specified (NOS) with respect to both lymph node status and recurrence-free survival. In multivariate analysis, the single most important factor for predicting recurrence free survival among the 207 patients was lymph node status. However, of 94 pure carcinomas, only 6 had lymph node metastases at the time of mastectomy. A detailed morphologic analysis demonstrated that two of these six cases were incorrectly diagnosed as being pure mucinous carcinomas--they were actually of the mixed type. In another two of these cases, the metastases originated from a co-existent infiltrating ductal carcinoma. Thus, metastases to the regional lymph nodes were observed in only two cases of pure mucinous carcinomas. It is concluded that the biologic behavior of the pure mucinous carcinomas differs from that of both the mixed carcinomas and infiltrating ductal carcinomas NOS. Thus, a new definition is suggested, in which a mucinous carcinoma is classified as such only if it is a pure carcinoma, and mixed mucinous carcinomas are classified according to the component without extracellular mucin production. PMID- 3028121 TI - Tissue polypeptide antigen--a marker antigen differentiating cholangiolar tumors from other hepatic tumors. AB - Twenty-two cases of primary hepatic tumors consisting of 11 hepatocellular carcinomas, 6 cholangiocarcinomas, 3 mixed hepatocellular and cholangiocellular carcinomas, and 2 biliary cystadenocarcinomas together with 8 cases of metastatic adenocarcinoma from various sites were studied by immunoperoxidase technic to demonstrate tissue polypeptide antigen. All of the tumors presumably derived from the epithelial lining of the bile duct, including cholangiocarcinoma, cholangiocarcinomatous portion of the mixed hepatocellular and cholangiocellular carcinoma, and biliary cystadenocarcinoma showed strong positive reaction. The hepatocellular carcinoma and the metastatic adenocarcinoma exhibited negative to weakly positive reactions. These results indicate that TPA can be of use in differentiating bile duct carcinomas from hepatocellular carcinoma and, to a lesser extent, from hepatic metastases of various adenocarcinomas. PMID- 3028122 TI - Koilocytosis preceding squamous cell carcinoma in situ of uterine cervix. AB - Association of koilocytic changes with cervical squamous cell carcinoma is well documented. The studies of such concurrent association may miss those cases of papillomavirus infection where cytomorphologic expression of virus has disappeared by the time carcinoma appears. The authors studied cytologic material before the diagnosis was made of cervical squamous cell carcinoma in situ in 25 patients. Twenty-two (88%) of the 25 patients showed koilocytosis compared with only 6 out of 57 (10.5%) in the control group. The findings of this study support a possible predisposing role of papillomavirus in squamous cell carcinoma of the cervix. PMID- 3028123 TI - Neuron-specific enolase. Assessment by ELISA in patients with small cell carcinoma of the lung. AB - Measurement of tissue-specific enolase isoenzymes may be of assistance in identifying small cell carcinomas of the lung and in distinguishing them from other pulmonary tumors. Enolase (E.C. 4.2.1.11) is a dimeric enzyme composed of various permutations of three immunologically distinct subunits alpha, beta, and gamma. Five isoenzymes alpha alpha, beta beta, gamma gamma, alpha beta, and alpha gamma have been identified. Immunohistochemical studies using antibodies to the gamma subunit have localized alpha gamma and gamma gamma specifically within neuronal and neuroendocrine tissues. Because of this limited distribution, neuron specific enolase (NSE) can function as a biochemical marker for neuroendocrine tumors. The authors developed an enzyme-linked immunosorbent assay (ELISA) using the double antibody sandwich method. The sandwich is composed of rabbit antirat enolase that cross-reacts to the human gamma monomer, making the test specific for the gamma gamma isoenzyme. The avidin-biotin-peroxidase complex system is used to provide increased assay sensitivity. Serum samples from patients with histologically diagnosed small cell carcinoma have concentration of NSE 20- to 30 fold greater than that found in normal serum. Studies were conducted on patients with a variety of malignant pulmonary lesions and compared with controls to determine the value of NSE as a tumor marker. PMID- 3028125 TI - Desmin immunoreactivity in glomus tumors. PMID- 3028124 TI - Angiotensin I-converting enzyme in a suprasellar germinoma. AB - High levels of angiotensin I-converting enzyme (ACE) were found by both direct radioimmunoassay and enzyme activity evaluation in a patient's plasma that had suprasellar germinoma. ACE levels returned to normal values after surgery, suggesting the tumor was the cause of the elevated plasma ACE. ACE was found in the tumor, and ACE activity was measured in a tumor extract. With the use of specific antihuman ACE antibodies, ACE was localized by immunohistochemistry in the tumoral germinal cells. Because ACE was also found in testicular germinomas, it appears to be a possible marker for germ cell tumors. PMID- 3028126 TI - Herpes simplex virus infection of the neonatal respiratory tract. AB - Three patients with neonatal herpes simplex virus (HSV) infection with upper or lower respiratory tract involvement are described. In all three patients, fever and respiratory tract symptoms were the presenting features and occurred in the absence of mucocutaneous lesions or maternal history of HSV Infection. In none of the children was HSV Infection considered likely at the time viral cultures were obtained. These patients demonstrate the need for consideration of HSV in the evaluation of neonatal respiratory tract disease, particularly if it is accompanied by fever and presents after the first several days of life. PMID- 3028127 TI - Cytomegalovirus infection and bronchopulmonary dysplasia in premature infants. AB - During a five-year period, 32 preterm infants weighing less than 2000 g were diagnosed as having postnatally acquired cytomegalovirus (CMV) infection in a neonatal intensive care unit. These CMV-infected infants were matched with 32 uninfected controls for gestational age, birth weight, and birth date; the two groups did not differ in Apgar scores or the incidence of respiratory distress syndrome and patent ductus arteriosus. Roentgenographic evidence of bronchopulmonary dysplasia (BPD) developed in 24 (75%) of 32 CMV-infected infants, an incidence significantly greater than that observed in control infants (12/32; 38%). Infants with acquired CMV infection required more respiratory support and longer hospitalization than uninfected controls. This association between acquired CMV infection in premature infants and the development of chronic lung disease provides further evidence that vigorous efforts to prevent CMV infection in hospitalized neonates is warranted. PMID- 3028128 TI - Adrenal function in thalassemia major following long-term treatment with multiple transfusions and chelation therapy. Evidence for dissociation of cortisol and adrenal androgen secretion. AB - Eight patients with beta-thalassemia who were given long-term treatment with combined multiple transfusions and chelation therapy underwent adrenal testing. The six male and two female patients ranged in age from 7 to 19 years. Six of eight patients had delayed bone ages and height greater than 2.5 SDs below the mean. Of the six patients more than 13 years of age, two had clinical evidence of isolated adrenarche and only one had evidence of true puberty. Cortisol levels were similar in patients and controls at zero time (10.6 +/- 1.8 micrograms/dL [292 +/- 50 nmol/L] vs 10.8 +/- 1.4 micrograms/dL [298 +/- 39 nmol/L]) and at 60 minutes (26.6 +/- 2.5 micrograms/dL [734 +/- 69 nmol/L] vs 24.9 +/- 1.9 micrograms/dL [687 +/- 52 nmol/L]) after insulin hypoglycemia (all values are the mean +/- SE). During an eight-hour infusion of ACTH, cortisol responses in the patients with thalassemia were not significantly different from those of controls. Baseline levels of the adrenal androgens dehydroepiandrosterone (DHEA) and dehydroepiandrosterone sulfate (DHEA-S) were significantly lower in the subjects with thalassemia compared with controls of similar bone age and pubertal status. The prolonged ACTH infusion caused a significant increase in the DHEA level (79.2 +/- 14.7 ng/dL [2.74 +/- 0.51 nmol/L] vs 538.6 +/- 38.1 ng/dL [18.67 +/- 4.79 nmol/L]) and the DHEA-S level (37.5 +/- 10.8 micrograms/dL [1.02 +/- 0.29 mumol/L] vs 70.5 +/- 18.3 micrograms/dL [1.19 +/- 0.50 mumol/L]) in the patients. The patients' peak stimulated levels of DHEA-S were significantly lower than those of the controls, whereas peak levels of DHEA were similar in the patients and the controls. These results indicate that combined multiple transfusions and chelation therapy preserve the integrity of the ACTH-cortisol axis in patients with thalassemia. The reduced levels of adrenal androgens, short stature, and delayed puberty noted in our patients suggest, however, that alternative approaches to the therapy of thalassemia are needed. PMID- 3028129 TI - Metastasis of hepatocellular carcinoma to the proximal jejunum manifested by occult gastrointestinal bleeding. AB - A 31-yr-old man, who had received left hepatic lobectomy for a ruptured hepatocellular carcinoma 17 months previously, began to have elevation of serum alpha-fetoprotein and occult gastrointestinal bleeding. Abdominal computed tomography, angiography, ultrasonography, upper gastrointestinal panendoscopy, colon-fiberscopy and endoscopic retrograde cholangiopancreaticography failed to disclose any recurrent intrahepatic tumor or bleeder in the gastrointestinal tract. However, a barium meal study revealed an intussusception with a suspicious nodular lesion in the proximal jejunum as the leading edge, which, after laparotomy, was proven to be a metastatic hepatocellular carcinoma and the cause of the gastrointestinal bleeding. The serosal side of this lesion was free from cancer involvement and there was no peritoneal implantation or lymph node involvement. The metastasis is probably hematogenous. The extremely unusual location involved by hepatocellular carcinoma in this patient signifies the ubiquitous nature of metastasis in this cancer. PMID- 3028130 TI - An epidemic of hepatitis A in an institution for young children. AB - A common-source epidemic of hepatitis A occurred in an Athenian institution boarding 38 children (mean age 4.8 years). All children were examined, and blood was drawn from each at the onset of the study and repeatedly during the next three months. Only one child (2.6%) was initially immune to hepatitis A virus as a result of prior infection. The attack rate (62.2%) and the ratio of icteric to anicteric cases (1:1.3) were high despite the administration of immunoglobulin (IG). Assays for anti-HAV IgM and a rising titer of anti-HAV IgG identified 19 (82.6%) and 22 (95.7%) of the 23 hepatitis A infections, respectively. One case (4.3%) was detected only by the presence of hepatitis A virus antigen and hepatitis A virus RNA in a fecal specimen, but these assays were otherwise marginally useful in this study. Nevertheless, the use of all available tests for the detection of hepatitis A virus is mandatory for the most accurate estimation of an epidemic of hepatitis A. Prompt administration of immunoglobulin had no effect on the number of clinical cases that were in the late incubation period, but it may have diminished the clinical expression of the infection and thus made diagnosis of infection more difficult. PMID- 3028131 TI - Effects of a 4-week freshwater fish (trout) diet on platelet aggregation, platelet fatty acids, serum lipids, and coagulation factors. AB - Eight healthy subjects consumed a diet in which all animal products were replaced by 750 g/day of freshwater trout. Platelet eicosapentaenoic acid (EPA) as a percentage of total platelet fatty acids rose from a prediet level of 0.2 +/- 0.4% to 3.4 +/- 1.6% after 4 weeks on the diet. Platelet arachidonic acid remained unchanged. Platelets became more hyperaggregable to collagen (p less than .025) but became hypoaggregable to arachidonic acid (p less than .05). The Ivy bleeding time became prolonged rising from a mean of 148 seconds before the diet to 168 seconds at 2 weeks and 202 seconds at 4 weeks. Serum lipids, coagulation profiles, and blood pressure remained unchanged. The changes induced by this diet were much less marked than changes induced by marine fish diets and diets supplemented by marine fish oil extracts, despite the fact that an equivalent amount of EPA was consumed and incorporated into platelets. These findings suggest that other substances, in addition to n-3 unsaturated fatty acids, may be responsible for the antithrombotic properties of marine fish. PMID- 3028132 TI - Restoration of impaired natural killer cell activity of B-chronic lymphocytic leukemia patients by recombinant interleukin-2. AB - Natural killer (NK) function in the majority of B-cell chronic lymphocytic leukemia patients is markedly deficient. This study was undertaken to determine if the biological response modifier interleukin-2 (IL-2), which is a potent augmenter of normal individuals' NK activity, could augment the low NK activity in these patients. Peripheral blood lymphocytes (PBL), depleted of B-cells, from most B-CLL patients exhibited low natural killer activity against NK-sensitive K562 cells and against herpes simplex virus (HSV)-infected lymphoblastoid cell lines (LCL). Incubation of patients' B-cell-depleted PBL with recombinant IL-2 resulted in augmentation of their NK activity against both K562 cells and HSV infected cells. Furthermore, whereas large granular lymphocytes (LGL) isolated from CLL patients are deficient in cytoplasmic granules, which are thought to play a role in NK-cell-mediated lysis, treatment of patients' LGL resulted in increased granulation by 4 hr after treatment with IL-2 and showed a concomitant increase in lytic activity comparable to that of normal individuals. PMID- 3028133 TI - Acetate dialysate versus bicarbonate dialysate: a continuing controversy. AB - The use of bicarbonate dialysate as the buffer during routine dialysis is growing. This discussion reviews several of the comparative trials in which bicarbonate and acetate buffers have been tested. Effects of the two buffers on BP, cardiac function, and pulmonary performance are discussed. Costs of the two systems are also compared. Patients who seem most likely to benefit from bicarbonate dialysate include those with a reduced muscle mass in whom a high sodium dialysate has not prevented hypotension. PMID- 3028134 TI - Abnormalities in the hypothalamic-pituitary-adrenocortical axis in patients with chronic renal failure. AB - The recently described human corticotropin-releasing factor was administered to eight patients with chronic renal failure in order to assess hypothalamic pituitary-adrenocortical (HPA) function. Acute administration of corticotropin releasing factor lead to a diminished increase of the basally elevated levels of ACTH and beta-endorphin immunoreactivity in patients on chronic hemodialysis. Basal plasma cortisol concentration was normal in end-stage renal disease; however, considering the corresponding elevated ACTH concentrations, cortisol levels were inadequately low. Thus, the hypothalamus as well as the adrenal gland seems to contribute to the alterations in HPA function observed in patients with chronic renal failure; involvement of the pituitary gland and effects of metabolic alterations cannot be ruled out. PMID- 3028136 TI - The morbidity and mortality of vermiculite miners and millers exposed to tremolite-actinolite: Part II. Mortality. AB - The vermiculite ore and concentrate of a mine and mill located near Libby, Montana was found to be contaminated with a fiber of the tremolite/acetinolite series. A study was conducted to estimate the exposure-response relationship for mortality for 575 men who had been hired prior to 1970 and employed at least 1 year at the Montana site. Individual cumulative fiber exposure (fiber-years) was calculated. Results indicated that mortality from nonmalignant respiratory disease (NMRD) and lung cancer was significantly increased compared to the U.S. white male population. For those workers more than 20 years since hire, the standard mortality rate (SMR) for lung cancer (ICDA 162-163) was 84.7, 225.1, 109.3, and 671.3 for less than 50, 50-99, 100-399, and more than 399 fiber-years respectively. Corresponding results for NMRD (ICDA 460-519) were 327.8, 283.5, 0, and 278.4. Based on a linear model for greater than 20 years since hire, the estimated percentage increase in lung cancer mortality risk was 0.6% for each fiber-year of exposure. At 5 fiber-years, the estimated percentage was 2.9% from an unrestricted (nonthreshold) linear model and 0.6% from a survival model. PMID- 3028135 TI - The morbidity and mortality of vermiculite miners and millers exposed to tremolite-actinolite: Part I. Exposure estimates. AB - The vermiculite ore and concentrate of a mine and mill near Libby, Montana, was found to be contaminated with fibrous tremolite-actinolite. Of 599 fibers (length greater than 5 microns and width greater than 0.45 micron) counted in eight airborne membrane filter samples, 96% had an aspect ratio greater than 10 and 16% had an aspect ratio greater than 50. Additionally, 73% of the fibers were longer than 10 microns, 36% were longer than 20 microns, and 10% were longer than 40 microns. Estimates of exposure before 1964 in the dry mill were 168 fibers/cc for working areas, 182 fibers/cc for sweepers, 88 fibers/cc for skipping, and 13 fibers/cc for the quality control laboratory. In 1964-1971, exposure estimates for these areas were 33, 36, 17, and 3 fibers/cc, respectively. Estimates of exposures in the mine before 1971 ranged from 9-23 fibers/cc for drillers and were less than 2 fibers/cc for nondrilling jobs. All 8-hr TWA job exposure estimates decreased from 1972-1976, and from 1977-1982 were less than 1 fiber/cc. PMID- 3028137 TI - The morbidity and mortality of vermiculite miners and millers exposed to tremolite-actinolite: Part III. Radiographic findings. AB - A study was conducted to estimate the exposure-response relationship for tremolite-actinolite fiber exposure and radiographic findings among 184 men employed at a Montana vermiculite mine and mill. Workers were included if they had been employed during 1975-1982 and had achieved at least 5 years tenure at the Montana site. Past fiber exposure was associated with an increased prevalence of parenchymal and pleural radiographic abnormalities. Smoking was not significantly related to the prevalence of small opacities. However, the number of workers who had never smoked was small, and this prevented measurement of the smoking effect. Under control for smoking and age, the prevalence of small opacities was significantly greater for vermiculite workers with greater than 100 fiber/cc-years exposure than for comparison groups (cement workers, blue collar workers, and coal miners) who had no known occupational fiber exposure. A logistic model predicted an increase of 1.3% in the odds ratio for small opacities at an additional exposure of 5 fiber-years. PMID- 3028139 TI - Silica-induced alveolar cell tumors in rats. AB - Min-U-Sil5, a form of alpha quartz, has been shown to induce peripheral lung tumors in rats exposed to the dust by inhalation. The animals were exposed to a nominal particle concentration of 12.4 mg/m3 for 8 hr/day, 4 days/week, for 2 years. The induced tumors were large and peripheral, and, when examined by electron microscopy, were found to be composed predominantly of alveolar type II cells. These cells were found in papillary, acinar, and solid forms of the tumors and were characterized by lamellar inclusion bodies. This is in contrast to the mouse, in which the papillary form was associated with Clara cells and the acinar form was linked with the type II cell. In this study, the Clara cell was a minor component of the tumor mass. No clear risk is established in man linking silica exposure to increased lung tumor rates. PMID- 3028138 TI - Asbestos exposure, cigarette smoking, and airflow limitation in long-term Canadian chrysotile miners and millers. AB - To investigate further the relationships of asbestos exposure, cigarette smoking, and airflow limitation, we have obtained detailed pulmonary function tests (PFT) in 331 long-term Canadian chrysotile workers, 34 of whom were lifetime nonsmokers. Three disease categories were defined on the bases of standard diagnostic criteria, gallium-67 lung uptake, and the lung pressure-volume curve. Category A was composed of workers without changes suggestive of alveolitis or asbestosis. There were eight nonsmokers (ns), among whom we found a statistically significant 30% reduction in airflow conductance (Gus) at low lung volume, which is consistent with the concept of an asbestos airway lesion. The 85 smokers (sm) of category A had reduction of Gus at both high and low lung volumes. Category B was composed of workers without asbestosis but with evidence of asbestos alveolitis. In the six ns, Gus was significantly reduced to 50% normal at low lung volume. The 59 sm had reduction of Gus at both high and low lung volumes but less severely than sm in category A. Category C was composed of workers with asbestosis. The 20 ns had restrictive pattern of lung function, and Gus was decreased to 39% normal at 50% TLC. The 153 sm in C had airflow reduction comparable to sm in B. We concluded that asbestos exposure, which leads to asbestos airway disease, asbestos peribronchiolar alveolitis, and asbestosis, causes airflow limitation at low lung volume but does not reduce the expiratory flow rates on the flow-volume curve in lifetime nonsmokers. In the smoking asbestos workers with alveolitis or asbestosis, the major component of airflow limitation is a smoking effect. In these smoking workers, rigidity of the lung lessens airflow obstruction associated with smoking at the expense of increased work of breathing. PMID- 3028141 TI - Mitral valve prolapse with symptoms of beta-adrenergic hypersensitivity. Beta 2 adrenergic receptor supercoupling with desensitization on isoproterenol exposure. AB - Autonomic nervous system dysfunction has recently been identified in a subset of patients with mitral valve prolapse. These autonomic nervous system abnormalities may correspond, in part, to biochemical alterations in beta-adrenergic receptors. Nine women with mitral valve prolapse and symptoms and signs of beta-adrenergic hypersensitivity and seven normal volunteer women were studied. Quiet standing (five minutes) increased both heart rate and plasma norepinephrine (p less than 0.05) in symptomatic patients with mitral valve prolapse compared with normal subjects. The dose of isoproterenol required either to increase heart rate 25 beats/minute (0.5 +/- 0.3 microgram versus 1.0 +/- 0.3 microgram) or to decrease mean arterial pressure 20 mm Hg (11.1 +/- 4.8 versus 78.2 +/- 25.2 micrograms) was significantly less in the patients with mitral valve prolapse than in the volunteers. Symptomatic patients with mitral valve prolapse were desensitized by a four-hour isoproterenol infusion, whereas sensitivity in normal control subjects did not change. In the patients with mitral valve prolapse, baseline beta-adrenergic receptor coupling was elevated compared with that in control subjects (220 +/- 7 versus 81 +/- 2; p less than 0.001). Isoproterenol infusion induced uncoupling in these patients (KL/KH = 35 +/- 3, p less than 0.05) but did not alter coupling in normal volunteers. This study demonstrates physiologic and pharmacologic beta-adrenergic hypersensitivity in vivo directly corresponding to biochemical supercoupling in a subset of patients with mitral valve prolapse. PMID- 3028140 TI - Cardiovascular effects of the aging process. AB - An understanding of the effects of aging upon the cardiovascular system assumes greater clinical importance as the population ages. Animal studies demonstrate prolonged tension development, impaired relaxation, and diminished response to receptor-mediated inotropic interventions in the cardiac muscle of senescent animals. Ventricular afterload is augmented at rest and increases during exercise through age-related alterations in peripheral arteries. Increased impedance to ejection may result in the modest concentric left ventricular hypertrophy seen in the elderly. Despite changes in cardiovascular function, resting and exercising cardiac outputs are well maintained in healthy elderly individuals who are free of occult coronary artery disease. The reduction in maximal oxygen consumption appears secondary to altered peripheral oxygen utilization rather than to cardiac causes. PMID- 3028142 TI - Congenital hypertrophy of the retinal pigment epithelium and familial polyposis of the colon. PMID- 3028143 TI - The mechanism of vascular leakage induced by leukotriene E4. Endothelial contraction. AB - This study identifies the microvascular target of leukotriene E4 (LTE4) by vascular labeling with carbon black and establishes the mechanism of its action at the cellular level by electron microscopy. LTE4 and its tripeptide precursor, leukotriene C4 (LTC4) were injected subcutaneously in guinea pigs. With LTE4, venular labeling was intense at 1000 and 100 ng and slight at 10 ng, with extinction at 1 ng. LTC4 induced a ring of labeled venules around a blank central area, suggestive of vasospasm. The nonpeptidyl leukotriene LTB4 induced no labeling. Histamine (1000 ng) induced an area of vascular labeling about equal to that by 1000 ng LTE4, but the labeling of individual venules was more intense. By electron microscopy, LTE4 was found to induce gaps in the endothelium of the venules; the endothelial cells adjacent to the gaps bulged into the lumen and showed wrinkled nuclei, consistent with cellular contraction. This ultrastructural evidence suggests that LTE4 increases vascular permeability by contraction of endothelial cells selectively, in the postcapillary venules, as was previously demonstrated for other inflammatory mediators, including histamine, serotonin, and bradykinin. PMID- 3028144 TI - Immunohistochemical localization of renin in renal tumors. AB - Immunoperoxidase staining for renin was performed with renal tumors, including juxtaglomerular (JG) tumor, Wilms' tumors, renal adenocarcinomas, renal oncocytomas, and cortical adenomas. Compared with the JG apparatus adjacent to the glomerulus, JG tumor cells were less darkly but diffusely stained for renin. One of five Wilms' tumors revealed more numerous renin-containing tumor cells than the adjacent renal cortex, whereas three of ten renal adenocarcinomas and two of three renal oncocytomas revealed only focally renin-positive tumor cell cytoplasms. None of six cortical adenomas were positive for renin. With available fresh tumor tissue, renin activity was studied by measuring newly formed angiotensin I by radioimmunoassay. JG tumor contained markedly elevated renin activity, whereas one Wilms' tumor and two renal adenocarcinomas contained no more than 2% of renin activity of the renal cortex, more than 50% of which was inactive renin. These findings suggest that the JG tumor elaborates enormous amounts of active renin, whereas other renal tumors produce lesser amounts of renin, more than half of which is inactive renin. PMID- 3028145 TI - Endosomal compartment of toad bladder epithelium. AB - Apical exocytosis and increased permeability are induced by antidiuretic hormone (ADH). After this, endocytosis is also induced by ADH and is associated with the decline in ADH-induced water permeability at the apical surface of the toad urinary bladder (9, 19, 20). During this process, horseradish peroxidase (HRP), a fluid phase marker, is taken up from the mucosal solution into endocytic tubules and multivesicular bodies. We now report that we can introduce from the apical (mucosal) side, a viral transmembrane protein (the G-protein of VSV) and that this protein can be retrieved as an integral membrane protein in endocytic membranes. This was demonstrated by immunoisolation of endosomal vesicles loaded with HRP using a monoclonal antibody against the cytoplasmic domain of the G protein. PMID- 3028146 TI - Adenylate cyclase regulation in intact cultured myocardial cells. AB - To examine the coupling of cardiac cell-surface beta-adrenergic receptors to adenylate cyclase activation and contractile response, we studied this receptor effector response system in monolayers of spontaneously contracting chick embryo ventricular cells under physiological conditions. The hydrophilic ligand 3H CGP12177 identified uniformly high-agonist affinity beta-adrenergic receptors (isoproterenol KD = 15 +/- 9 nM). Isoproterenol-stimulated cyclic AMP (cAMP) accumulation with 50% effective concentration at (EC50) = 12.1 nM and augmented contractile response with EC50 = 6 nM under identical conditions. One micromolar isoproterenol induced receptor loss from the cell surface with t1/2 = 13.2 min; under identical conditions cAMP content declined with t1/2 = 13.5 min and contractile response with t1/2 = 20.7 min. After agonist removal cAMP response recovered with t1/2 = 15.7 min and receptors with t1/2 = 24.7 min. Sixty minutes after agonist removal there was recovery of 52% of maximal cAMP responsiveness and 82% of the initial number of receptors; receptor occupancy was associated with 78% of initial contractile response. Agonist affinity for cell-surface receptors was changed only modestly by agonist exposure. We conclude that for this system there is relatively close coupling between high-affinity receptors, adenylate cyclase stimulation, and contractile response. PMID- 3028147 TI - Sodium-hydrogen exchange system in LLC-PK1 epithelium. AB - Sodium influx into LLC-PK1 cells has been characterized. The main amiloride sensitive pathway for sodium entry through the apical membrane in this cell line is sodium-hydrogen exchange with an apparent Km for sodium of 19 mM. Influx is pH dependent and is amiloride sensitive with K1/2 of 30 microM. Inhibition of the sodium transport by protons (at 1 mM external sodium) is consistent with an interaction of H+ ions at a single site having an apparent pKH of 7.2. These data are similar to those reported previously for brush border membrane (BBM) isolated from kidney proximal tubule. However, in contrast to previously published reports, H+ does not compete with amiloride in blocking Na+ influx and exhibits a mixed inhibition with sodium ions. The latter, together with a direct and independent assessment of amiloride and sodium interaction, indicates that amiloride does not compete with sodium for the same site. Possible explanations for the discrepancy in the literature concerning the interaction of Na+, H+, and amiloride with the Na+-H+ exchanger and the characteristics of the Na+-H+ exchange system in LLC-PK1 are discussed. PMID- 3028148 TI - Specificity of effect of osmolality on aldosterone secretion. AB - Small (3-7 mM) changes in [NaCl] have a marked inverse effect on angiotensin II- or K-stimulated aldosterone secretion by the isolated, perfused canine adrenal gland. The effect is due to the accompanying changes in osmolality rather than to the changes in [Na] or [Cl]. The present study was undertaken to determine whether osmolality is a specific and discrete signal that modulates the secretion of aldosterone only or is simply a nonspecific physical factor that alters the secretion of other adrenocortical hormones as well. The study also determined whether small changes in osmolality affect the conversion of corticosterone to aldosterone. The secretion of both cortisol and aldosterone by isolated canine adrenal glands responded in a dose-dependent fashion to adrenocorticotropin (ACTH), but in contrast to the rapid and potent modulating action of osmolality reported previously for angiotensin II- or K-stimulated aldosterone secretion, changes in osmolality at the midpoint of ACTH infusion had no detectable effect on either cortisol secretion or, unexpectedly, aldosterone secretion. This indicates that osmolality is a highly specific signal that modulates responsiveness of the zona glomerulosa to the factors, angiotensin II and K, which are considered to be most important in the acute regulation of aldosterone secretion, but does not influence secretion of cortisol by inner zones of the adrenal cortex. In glands treated with agents that block aldosterone production from endogenous precursors, small changes in osmolality had no detectable effect on the conversion of exogenous corticosterone to aldosterone.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3028149 TI - Effects of forskolin on bone in organ culture. AB - The effects of forskolin, which directly activates adenylate cyclase in most systems, have been compared with the actions of parathyroid hormone and calcitonin, both of which have been suggested to utilize cAMP as a second messenger in their actions on bone. Forskolin alone stimulated calcium release from neonatal mouse calvaria and fetal rat limb bones in vitro in a dose dependent manner. The effect was maximal at 10(-6) M in both systems. At higher concentrations forskolin completely inhibited stimulated bone resorption, although with submaximal concentrations the inhibition was only partially sustained up to 72 h. Forskolin directly stimulated cAMP release from calvaria into the medium at concentrations up to 10(-4) M. Forskolin had no effect on the interaction between parathyroid hormone and calcitonin, while calcitonin inhibited the stimulatory effect of forskolin comparably with its inhibition of parathyroid hormone-stimulated bone resorption. The results indicate that forskolin has dual effects on bone and can mimic responses of both parathyroid hormone and calcitonin in both bone culture systems. The observed response depends on the concentration of forskolin used and the length of treatment with the drug. PMID- 3028151 TI - Na-H exchange regulates intracellular pH in isolated rat hepatocyte couplets. AB - Intracellular pH (pHi) was measured directly in isolated rat hepatocyte couplets using pH sensitive microelectrodes. The hepatocytes were maintained in a minimal salt buffer without added hormones or serum. Values of pHi (6.99 +/- 0.12, mean +/- SE) were close to their Nernst equilibria. After intracellular acidification with ammonium chloride, pH regulation was inhibited with 1 mM amiloride or by omission of external sodium, consistent with a Na-H exchange mechanism. Mean intracellular buffering power, in the nominal absence of carbon dioxide, was 34.1 +/- 11.4 mM. In the presence of external bicarbonate, amiloride or omission of sodium slowed, but did not completely inhibit recovery from acidification, indicating that additional pHi regulation mechanisms may operate in this preparation. These studies provide a direct measurement of pHi in hepatocyte couplets and indicate that Na-H exchange, together with a bicarbonate dependent system are important mechanisms for pHi regulation in this preparation. PMID- 3028150 TI - Maitotoxin stimulates steroid and peptide hormone secretion by swine luteal tissue. AB - The impact of calcium influx on the secretion of steroidal and nonsteroidal ovarian products was tested using a novel marine toxin, maitotoxin, a putative activator of endogenous calcium channels. In incubations of swine luteal minces in balanced salt solution, maitotoxin induced dose-dependent (3-12.5 ng/ml) and time-dependent (15-60 min) secretion of progesterone and relaxin. The magnitude of these stimulatory effects approached that observed for luteinizing hormone (LH) and was explicitly dependent on the presence of extracellular calcium ions. Maitotoxin's actions were not significantly impeded by substitution of choline for sodium ions in the incubation medium. Moreover, measurements of tissue calcium content by atomic absorption spectrometry indicated that maitotoxin augmented the total cellular content of calcium by 44 +/- 3.1% within 1 min. This time course was distinct from that observed for LH. The specificity of these actions on the calcium pathway was further suggested by the absence of any measurable effects of maitotoxin on luteal cAMP generation. We conclude that maitotoxin transiently increases luteal calcium content without altering cAMP generation and elicits concomitant release of both a steroidal (progesterone) and nonsteroidal (relaxin) product of the corpus luteum. Such actions are dependent on extracellular calcium but not sodium ions. These observations provide evidence for a calcium-dependent stimulatory pathway that activates both steroidal and nonsteroidal mechanisms of secretion in the swine corpus luteum. PMID- 3028152 TI - Ontogeny of rat gastric H+-K+-ATPase activity. AB - Gastric H+-K+-ATPase activity was examined in subcellular fractions (P1, 1,000 g; P2, 20,000 g; P3, 100,000 g) obtained from fundic mucosae of fetal, newborn, and adult rats. Depending on the tissue secretory state the distribution of this enzyme activity predominates in P3, i.e., resting adult, or in P2, i.e., stimulated adult, while total enzyme activity (P2 + P3) remains constant. Enzyme activity was detected as soon as day 19 of gestation and P2 + P3 reached 2.2 +/- 0.2 mumol Pi X mg prot-1 X h-1 at parturition. H+-K+-ATPase activity steadily decreased from 4.0 +/- 0.7 mumol Pi X mg prot-1 X h-1 on day 6 after birth to approximately 0 on day 12. This activity recovered as soon as day 13 (3.5 +/- 0.6 mumol Pi X mg prot-1 X h-1) and continued to rise slowly until adulthood. Incubation of subcellular fractions with 0.1 mM omeprazole resulted in a total loss of H+-K+-ATPase activity at all the stages of development examined. These studies of functional relationship during the development between gastric acid secretion and distribution of gastric H+-K+-ATPase activity among P2 and P3 fractions indicate that this last could be considered as an index of the secretory state of the parietal cell. This enzyme's pattern of development in P2 + P3, correlated with the one previously observed for stimulated H+ secretion in intact tissue, suggest that the H+-K+-ATPase is the limiting step in response to secretagogues during postnatal period. PMID- 3028153 TI - Specific alpha 1-, alpha 2-, and beta-responses to norepinephrine in pyruvate perfused rat kidneys. AB - We compared the effects of alpha- and beta-adrenergic agonists on vascular resistance, gluconeogenesis (GNG), and electrolyte excretion in pyruvate-perfused rat kidneys. Norepinephrine (NE) (15-60 nM) increased vascular resistance (2-19%) and GNG (42-49%) and decreased fractional excretion of Na (18-34%), Cl (38-48%), and K (13-27%). Prazosin and yohimbine blocked the vasoconstrictor but not the antinatriuretic response. Prazosin revealed an antiphosphaturic and inhibited the gluconeogenic response. Propranolol blocked the effect on Na and Cl (66%) and K excretion (100%). Methoxamine (0.6-1 microM) maximally increased GNG (40-52%) with little effect on vascular resistance (+0-9%) and NaCl excretion (-0-11%). Adding glucose to the perfusate increased methoxamine's effects on electrolyte excretion and vascular resistance. Mercaptopicolinate did not alter methoxamine's effects during pyruvate perfusion. Clonidine (0.1 microM) decreased phosphate excretion without vasoconstriction or antinatriuresis; 0.5-1 microM vasoconstricted and decreased NaCl excretion. Isoproterenol (0.1 microM) vasodilated and decreased Na (25%), Cl (40%), and K excretion (20%). Thus alpha 1 and beta-receptors could account for the effect of NE on GNG and K reabsorption, respectively. Synergism of alpha 1, alpha 2, and beta were required for full antinatriuresis. Glucose increased alpha 1-stimulated vasoconstriction and antinatriuresis. Blocking alpha 1-receptors revealed an alpha 2-like antiphosphaturic response. PMID- 3028154 TI - cAMP-associated inhibition of Na+-H+ exchanger in rabbit kidney brush-border membranes. AB - Adenosine 3',5'-cyclic monophosphate (cAMP) inhibits the rate of bicarbonate reabsorption and the rate of Na+-H+ exchange transport in the apical membrane of the proximal convoluted tubule. To study the relation between cAMP, cAMP dependent protein kinase, and Na+-H+ exchange transport, brush-border membrane vesicles from the rabbit kidney were phosphorylated in vitro. The rate of proton gradient-stimulated amiloride-inhibitable 22Na+ uptake was measured as an index of Na+-H+ exchange transport activity. The inclusion of cAMP (10(-6) M) in a phosphorylating solution containing ATP decreased the 10-s uptake of amiloride sensitive sodium from 2.25 +/- 0.21 nmol/mg protein in controls to 1.94 +/- 0.19 (P less than 0.001). Incubation of vesicles in the presence of purified catalytic subunit of cAMP-dependent protein kinase inhibited the amiloride-sensitive uptake of 22Na+ at 10 s from 2.35 +/- 0.49 nmol/mg protein to 2.05 +/- 0.44 (P less than 0.005). The inhibitory effect of both cAMP and catalytic subunit of cAMP dependent protein kinase was blocked by the specific thermostable protein inhibitor of the kinase. These studies demonstrate that activation of endogenous membrane-bound cAMP-dependent protein kinase or exposure to exogenous catalytic subunit of cAMP-dependent protein kinase inhibits the rate of Na+-H+ exchange transport in the brush-border membrane of the rabbit kidney. PMID- 3028155 TI - Dopamine causes inhibition of Na+-K+-ATPase activity in rat proximal convoluted tubule segments. AB - We studied the effect of dopamine (DA) on Na+-K+-ATPase activity in proximal convoluted tubule (PCT) segments dissected from perfused rat kidneys. DA inhibited Na+-K+-ATPase activity in a dose-dependent manner. Inhibition was significant with 10(-7) M DA and maximal with 10(-4) M DA. The inhibition was reversible. Enzyme inhibition occurred in the presence of DA and a DA antagonist, metoclopramide, but not when 10(5) M of the DA1 and DA2 agonists fenoldopam mesylate and LY 171555 were added in the absence of DA. In PCT segments incubated with the DA precursor dopa, Na+-K+-ATPase activity was also inhibited. However, dopa did not inhibit the sodium pump if dopa decarboxylase activity was blocked with benserazide. These findings suggest an intracellular site of action of DA. In tubules incubated in different K concentrations, 10(-5) DA decreased the maximal activity (Vmax) and increased the Km. DA 10(-5) M caused an almost immediate swelling of PCT segments. Swelling did not occur in the presence of both DA and 10(-5) M amiloride. The DA-induced tubular swelling was probably due to inhibition of Na+-K+-ATPase-mediated Na+-transport. We conclude that in rat PCT segments, DA causes a rapid and reversible inhibition of apparent Na+-K+ ATPase activity and an apparent reduction in the affinity for K. The site of action appears to be intracellular. PMID- 3028156 TI - Adenosine inhibits beta-adrenoceptor but not DBcAMP-induced renin release. AB - The purpose of these studies was to assess the direct effect of adenosine on the renin release response to beta-adrenoceptor activation in vivo in the canine kidney. In an initial study, innervated, intrarenal beta-adrenoceptors were activated selectively via renal nerve stimulation in kidneys in which the alpha adrenoceptor response to renal nerve stimulation had been blocked with phentolamine. Adenosine, infused directly into the renal artery (10 and 30 micrograms/min), significantly blunted the renin release response to renal nerve stimulation. However, adenosine also caused significant reductions in base-line glomerular filtration rate, sodium excretion rate, and filtration fraction. To eliminate these confounding effects of adenosine on renal function and to prevent changes in norepinephrine release due to prejunctional inhibition by adenosine, we also studied the effect of intrarenal infusions of adenosine on norepinephrine induced renin release in the nonfiltering, alpha-adrenoceptor-blocked, canine kidney. In this model of beta-adrenoceptor activation, adenosine abolished the renin release response to intrarenal infusions of norepinephrine. In a final series of experiments, the effect of adenosine on the renin response to dibutyryl adenosine 3',5'-cyclic monophosphate (cAMP) was examined in the nonfiltering, beta-adrenoceptor-blocked, canine kidney. In this model of cAMP-induced renin release, adenosine was ineffective in attenuating the renin release response. These data demonstrate that in vivo adenosine directly inhibits beta-adrenoceptor mediated renin release by a mechanism that does not involve a reduction in the ability of cAMP to activate intracellular mechanisms leading to renin release. PMID- 3028157 TI - Atrial natriuretic peptide stimulates salt secretion by shark rectal gland by releasing VIP. AB - Salt secretion by the isolated perfused rectal gland of the spiny dogfish shark, Squalus acanthias, is stimulated by synthetic rat atrial natriuretic peptide (ANP II) as well as extracts of shark heart, but not by 8-bromo-cyclic guanosine 5' monophosphate. Cardiac peptides have no effect on isolated rectal gland cells or perfused tubules, suggesting that stimulation requires an intact gland. The stimulation of secretion by ANP II is eliminated by maneuvers that block neurotransmitter release. These include: perfusion with procaine (10(-2) M), perfusion with high Mg2+ (9.5 mM) and low Ca2+ (0.5 mM) concentrations, and addition to the perfusate of the calcium channel blockers nifedipine (10(-6)M), diltiazem (5 X 10(-5)M), or verapamil (10(-4)M). Cardiac peptides stimulate the release of vasoactive intestinal peptide (VIP), known to be present in rectal gland nerves, into the venous effluent or perfused glands in parallel with their stimulation of salt secretion, but the release of VIP induced by ANP II is prevented by perfusion with procaine. Cardiac peptides thus appear to regulate rectal gland secretion by releasing VIP from neural stores within the gland. It is possible that other physiological effects of these hormones might be explained by an action to enhance local release of neurotransmitters. PMID- 3028158 TI - Kinetic study of Na+-K+ pump in erythrocytes from essential hypertensive patients. AB - The interaction of the Na+-K+ pump with internal Na+ was investigated in erythrocytes from 38 normotensive control subjects and 49 essential hypertensive patients. In six of the hypertensive patients, the Na+-K+ pump exhibited an apparent dissociation constant for internal Na+ (KNa) above an upper normal limit of 7 mmol/l cells. Four of these six hypertensives showed an increase in the maximal rate of ouabain-sensitive Na+ efflux (Vmax), above an upper normal limit of 11 mmol X l cells-1 X h-1. These abnormalities were stable in repeated determinations over 1-3 yr. A kinetic study of other erythrocyte Na+ transport pathways showed that 16 hypertensives had a low apparent affinity of the Na+-K+ cotransport system for internal Na+, 10 hypertensives exhibited increased Na+-Li+ countertransport fluxes, and 11 hypertensives had increased Na+ leak. None of these three abnormalities were observed in the six hypertensives with abnormal pump fluxes. We thus propose to denominate them as Pump (-) hypertensives. Interestingly, four Pump (-) hypertensives exhibited an increased maximal rate of outward Na+-K+ cotransport. Basal erythrocyte Na+ content of Pump (-) hypertensives was within normal range. This suggests that the increased maximal rates of the Na+-K+ pump and Na+-K+ cotransport system compensate the low pump affinity for internal Na+. PMID- 3028159 TI - Inorganic phosphate inhibits sympathetic neurotransmission in canine saphenous veins. AB - Inorganic phosphate has been proposed as the initiator of metabolic vasodilatation in active skeletal muscle. The present study was primarily designed to determine if this substance has an inhibitory effect on adrenergic neurotransmission. Rings of canine saphenous veins were suspended for isometric tension recording in organ chambers. A comparison was made of the ability of inorganic phosphate (3 to 14 mM) to relax rings contracted to the same degree by electrical stimulation, exogenous norepinephrine, and prostaglandin F2 alpha. The relaxation during electrical stimulation was significantly greater at all concentrations of phosphate. In strips of saphenous veins previously incubated with [3H]norepinephrine, the depression of the contractile response caused by phosphate during electrical stimulation was accompanied by a significant reduction in the overflow of labeled neurotransmitter. Thus inorganic phosphate inhibits sympathetic neurotransmission and hence may have a key role in the sympatholysis in the active skeletal muscles during exercise. By contrast, in this preparation, it has a modest direct relaxing action on the vascular smooth muscle. PMID- 3028160 TI - Two types of transient outward currents in adult human atrial cells. AB - It has been suggested in a previous article [Escande et al., Am. J. Physiol. 249 (Heart Circ. Physiol. 18): H843-H850, 1985] that transient outward currents may participate in the initial repolarization of human atrial fibers. The present study substantiates the existence of such currents in human myocardium. Membrane currents were recorded in enzymatically dissociated cells using the whole cell patch-clamp technique. Two kinds of transient outward currents were observed: 1) a long-lasting outward current, (ilo), which was suppressed by 4-aminopyridine. The time to peak of ilo was 18.0 +/- 0.7 ms, and its inactivation time constant was 35.7 +/- 2.1 ms at room temperature (test pulses, +20 mV; holding potential, 40 mV); 2) a brief outward current (ibo), which persisted with 3 mM 4 aminopyridine and exhibited a shorter time to peak (5.5 +/- 0.2 ms) and a faster decay (time constant, 9.1 +/- 1.8 ms). ilo was inhibited by Ba but was insensitive to the calcium blocker Co. Co blocked both the slow inward current (isi) and ibo. It is concluded that two different transient outward currents control the repolarization in human atrial cells. PMID- 3028161 TI - mu-Opioid receptors in NTS elicit pressor responses via sympathetic pathways. AB - We have evaluated the relative contributions of the sympathetic and parasympathetic nervous systems to the increased mean arterial pressure (MAP) and heart rate (HR) elicited by the selective mu-agonist D-Ala2, MePhe4, Gly-ol5 enkephalin (DAGO) following microinjection (100 nl) into the nucleus of tractus solitarius (NTS) of anesthetized, artificially ventilated Sprague-Dawley rats. The effects of anesthesia and central opioid-receptor activation on baroreflex function were also examined. All cardiovascular responses elicited by DAGO were eliminated by complete C1 spinal transection. Pretreatment with the alpha adrenergic antagonist phentolamine attenuated the increase in MAP, but not the tachycardia; the beta-blocker propranolol abolished the tachycardia but not the pressor response to DAGO. Adrenalectomy, vagotomy, or pretreatment with atropine methyl nitrate were all without effect. Baroreflexes were attenuated in animals anesthetized with pentobarbital sodium, but were present in urethan-anesthetized rats. DAGO attenuated the increases in MAP and HR elicited following carotid occlusion, but not the bradycardia elicited by a phenylephrine-induced pressor response. These data indicate that mu-receptors in the NTS elicit cardiovascular responses that are mediated by increased sympathetic nerve activity, and accompanied by selective attenuation of baroreflex function. PMID- 3028162 TI - Differentiation of sarcoplasmic reticulum during cardiac myogenesis. AB - The composition and function of fetal and mature sheep cardiac sarcoplasmic reticulum membranes were investigated. Phospholamban, a major phosphoprotein in the mature sarcoplasmic reticulum membranes, was present in early stages of cardiac myogenesis. This fetal form of phospholamban was phosphorylated by cAMP dependent protein kinase but not in the presence of Ca2+ and calmodulin. Ca2+ uptake and Ca2+-dependent ATPase activity were low in fetal sarcoplasmic reticulum compared with the adult controls, although the apparent affinities for Ca2+ were similar. Sarcoplasmic reticulum vesicles isolated at all developmental stages had very low levels of plasma membrane (as determined by Na+-K+-ATPase and Na+-Ca2+ exchanger activities) and mitochondrial contamination. Sarcoplasmic reticulum Ca2+ uptake and Ca2+-dependent ATPase activities were not affected by micromolar concentrations of vanadate, and the accumulated Ca2+ could not be released by the addition of NaCl. The amount of both the 110- and 55-kDa protein bands, identified with specific antibodies as Ca2+-ATPase and calsequestrin, respectively, was low in early stages of cardiac myogenesis. Age-related differences in the Ca2+ transport properties of cardiac sarcoplasmic reticulum and in the amount of the Ca2+-ATPase and calsequestrin may explain alterations in the regulation of intracellular Ca2+ concentrations in the fetal heart. This may contribute to the developmental changes in myocardial function. PMID- 3028163 TI - Deinhibition of cardiac Na+-K+-ATPase after exposure to exogenous phospholipase A2. AB - After 2 h of exogenous phospholipase A2 (PLA2) exposure, membrane phospholipid decreased from 3.22 +/- 0.31 to 1.06 +/- 0.13 mumol/mg (33% of control). All classes of phospholipid, except sphingomyelin, were hydrolyzed, whereas total cholesterol content was unaffected. Increases in nonesterified fatty acids (NEFA) were reflected primarily in oleic (18:1), linoleic (18:2), and arachidonic (20:4). Na+-K+-adenosinetriphosphatase (ATPase) activity was inhibited to 29% of control by 2 h of PLA2 treatment, and this inhibition was reversed (albeit, not completely after 5 min of PLA2 treatment) by removal of the hydrolysis products with 0.1% bovine serum albumin (BSA). In contrast, the apparent binding capacity for [3H]ouabain was not affected by PLA2 treatment. Unmasking of latent [3H]ouabain binding by alamethicin was utilized to estimate changes in the proportion of sealed vesicles present before and after PLA2 treatment. PLA2 treatment resulted in a time-dependent loss of sealed vesicles that paralleled the time course of phospholipid hydrolysis and was not reversed by washing with BSA. These studies demonstrate that cardiac Na+-K+-ATPase activity is inhibited by accumulation of endogenously produced lysophospholipids and NEFA. In contrast, loss of vesicle integrity may result from both accumulation of endogenously produced hydrolysis products and membrane phospholipid depletion. PMID- 3028164 TI - Factors from paraventricular nucleus mediating adrenocorticotropin release in cats. AB - Experiments were conducted in alpha-chloralose-urethan-anesthetized cats. We stimulated the hypothalamic paraventricular nucleus (PVH) electrically before and after intracerebroventricular (icv) injection of an antiserum to arginine vasopressin (AVP), one to corticotropin-releasing factor (CRF), or one to normal rabbit serum (NRS). Stimulation of the ventral portion of the dorsal PVH led to increases in arterial pressure and heart rate that did not change after any of the icv treatments. However, the effect of each agent on the increases in plasma adrenocorticotropin (ACTH) after stimulation was related to the area of the PVH that was stimulated. The response to stimulation of a rostral area that extended dorsally from the anterior PVH was blocked completely by the anti-AVP but not by anti-CRF or NRS. The response to stimulation of a caudal area located in the dorsal PVH was attenuated after anti-CRF, unchanged after anti-AVP, and augmented after NRS. The antibodies that were given icv were found immunocytochemically to enter the median eminence at sites that include some adjacent to the portal vessels. Immunocytochemical localization of AVP- and of CRF-containing neurons in the PVH showed that the anterior PVH had the highest proportion of AVP neurons in the PVH but had only a few CRF neurons. In contrast, the dorsal PVH contained the highest density of CRF neurons in the PVH as well as some AVP neurons. We suggest that, in the cat, the primary releasing factor for the anterior PVH is AVP and that for the dorsal PVH is CRF. PMID- 3028165 TI - Glucagon stimulation of brown adipose tissue growth and thermogenesis. AB - Despite long-standing observations of a whole-body thermogenic effect of glucagon, the role of glucagon in activating thermogenesis in brown adipose tissue has not often been studied. We investigated the ability of administered glucagon to produce alterations in brown adipose tissue similar to changes produced by accepted stimuli of brown fat activity: cold, norepinephrine, and overfeeding. Eighteen days of glucagon injections (1 mg/kg) to male Sprague Dawley rats produced, relative to saline-injected controls, decreases in feed efficiency and increases in brown adipose tissue weight, protein content, DNA content, and mitochondrial mass as reflected in cytochrome oxidase activity. The observed changes were similar, though of lesser magnitude, to changes produced in these same parameters induced by administration of norepinephrine (250 micrograms/kg) for a positive control group. Four days of glucagon administration (1 mg/kg) produced increases in specific activity of cytochrome oxidase and lipoprotein lipase. After 8 days of glucagon administration, changes in whole-pad activity similar to those seen with 18 days of administration were present. Glucagon also increased whole-pad lipoprotein lipase activity after 4 and 8 days. Surgically denervated interscapular brown adipose tissue retained its ability to respond to exogenous glucagon, though the magnitude of the response was diminished. Guanosine 5'-diphosphate (GDP) binding to brown adipose tissue mitochondria was measured as an assessment of functional state after 5 days of glucagon (1 mg/kg). There was an increase in GDP binding relative to controls whether expressed as picomoles per milligram mitochondrial protein or nanomoles per pad.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3028166 TI - Pressoreceptor modulation of renal but not splenic sympathetic reflexes. AB - Influences of sinoaortic and vagally innervated vascular pressoreceptors on excitatory splenic and renal sympathetic responses to splenic receptor stimulation were investigated in anesthetized cats. These experiments demonstrated that these pressoreceptors have little apparent effect on the magnitude of splenic nerve responses to splenic receptor stimulation by capsaicin, bradykinin, or congestion. In contrast, activation of these pressoreceptors attenuated renal nerve responses to splenic receptor stimulation. Influences of sinoaortic and vagally innervated receptors on tonic sympathetic nerve activity also were evaluated. Stimulation of these receptors by small increases in arterial pressure (15-21 mmHg) caused equivalent inhibition of splenic and renal nerve activity; large increases (50-66 mmHg) caused significantly greater inhibition of renal than splenic nerve activity. These results illustrate that excitatory renal and splenic sympathetic responses to splenic receptor stimulation are not suppressed equally by pressoreceptor activation, vascular pressoreceptors can have greater inhibitory influences on tonic renal than splenic nerve activity, and vascular pressoreceptor influences on sympathetic reflexes are similar to those on tonic nerve activity. PMID- 3028167 TI - Na+-H+ exchange and Na+-dependent transport systems in streptozotocin diabetic rat kidneys. AB - The streptozotocin-induced diabetic rat was used to test the hypothesis that Na+ H+ exchange activity in the proximal tubule luminal membrane would be increased in association with renal hypertrophy, altered glomerular hemodynamics, enhanced filtered load and tubular reabsorption of Na+, and stimulated Na+ pump activity in the basolateral membrane, previously reported characteristics of this experimental animal model. Amiloride-sensitive H+ gradient-dependent Na+ uptake and Na+ gradient-dependent H+ flux were increased in brush-border membrane vesicles from the streptozotocin-treated animals. Na+ gradient-dependent uptakes of phosphate, D-glucose, L-proline, and myoinositol were decreased in the drug induced diabetic animals. These membrane transport alterations were not found when the streptozotocin-diabetic animals were treated with insulin. PMID- 3028168 TI - Renal alpha 2-adrenoceptors and the adenylate cyclase-cAMP system: biochemical and physiological interactions. AB - alpha 2-Adrenoceptors were first described pharmacologically ten years ago. Within three years their capacity to inhibit adenylate cyclase had been demonstrated in many tissues. They were demonstrated biochemically in the kidneys in 1981 even before any renal physiological effects of their activation were known. They predominate numerically over alpha 1-adrenoceptors in renal membranes and their density is increased in genetic forms of rat hypertension. alpha 1 Adrenoceptors normally mediate the vasoconstriction and sodium- and water retaining effects of sympathetic neuronally released norepinephrine. Norepinephrine or epinephrine must be infused to activate alpha 2-adrenoceptors, suggesting that renal alpha 2-adrenoceptors are extrajunctional, whereas alpha 1 adrenoceptors are postjunctional. When alpha 1-adrenoceptors are chronically blocked, renal alpha 2-adrenoceptor density increases and they assume a location at postjunctional sites, the otherwise exclusive domain of alpha 1-adrenoceptors. Results from microdissection studies have established that alpha 2-adrenoceptors are present on most segments of the nephron and that their activation can suppress adenosine 3,'5'-cyclic monophosphate (cAMP) accumulation induced by most renal hormones. However, failure of alpha 2-adrenoceptor activation to suppress cAMP accumulation in some tubular segments induced by certain hormones suggests compartmentalization of adenylate cyclase regulation that is hormone-function specific. In view of the potent inhibitory effects of alpha 2-adrenoceptor stimulation on hormone activated cAMP accumulation in several discrete areas of the nephron, we suggest that alpha 2-adrenoceptors fulfill important regulatory role(s) in renal function. To date, alpha 2-adrenoceptor activation has been shown to reverse vasopressin-induced sodium and water retention, and arachidonic acid- and furosemide-induced cAMP, sodium, and water excretion in the isolated perfused kidney. Thus the effects are qualitatively and quantitatively dependent in these studies on the hormone being infused and are therefore hormone-function specific. Physiological effects of alpha 2-adrenoceptor activation of thyrocalcitonin and on parathyroid hormone-induced effects have not been studied. alpha 2-Adrenoceptor activation can inhibit renin release in some model systems and can activate a sodium-hydrogen antiporter system in proximal tubules. The physiological roles of these actions are unknown. PMID- 3028169 TI - Norepinephrine increases Na+-K+-ATPase and solute transport in rabbit proximal tubules. AB - We studied the effects of norepinephrine on solute transport and oxidative metabolism in proximal tubules. Norepinephrine (10(-6) M) in the bath stimulated fluid absorption (Jv) by proximal convoluted tubules from 0.76 +/- 0.10 to 1.01 +/- 0.11 nl X mm-1 X min-1 (P less than 0.001). Bicarbonate, chloride, and phosphate transport also increased in proportion to the increases in Jv. Norepinephrine increased ouabain-sensitive oxygen consumption (QO2) in suspensions of cortical tubules by 1.3 nmol X mg protein-1 X min-1 and had no effect on ouabain-insensitive QO2 or mitochondrial respiration. Na+-K+-ATPase activity in basolateral membranes prepared from cortical homogenates incubated with norepinephrine increased from 277.1 +/- 34.9 to 411.1 +/- 38.6 nmol Pi X mg protein-1 X min-1 (P less than 0.005). Norepinephrine also increased Na+-K+ ATPase activity of cortical homogenates by 72% but had no effect on Na+-K+-ATPase if added directly to purified basolateral membranes. These studies show that norepinephrine stimulates solute transport in the proximal tubule by increasing Na+-K+-ATPase activity indirectly through some component or components of the adrenergic receptor system. PMID- 3028170 TI - Adaptation to metabolic acidosis by turtle urinary bladder. AB - We utilized the turtle urinary bladder to study the mechanisms responsible for adaptation to metabolic acidosis. Bladders removed from acidotic turtles had a higher rate of H+ secretion in vitro than bladders from control turtles, despite identical extracellular pH. HCO3 secretion, however, was not different between the two groups. The increase in H+ secretion could be mediated by a decrease in intracellular pH and/or by an increase in the number of cells thought to be responsible for H+ secretion. To study this issue, we measured intracellular pH with the fluorescent dye 6-carboxyfluorescein diacetate and quantified the number of cells by fluorescence microscopy utilizing acridine orange, rhodamine 123, and 6-carboxyfluorescein diacetate in turtles receiving different acid loads. Urinary acidification measured in vivo was increased in turtles fed a low-acid load for 48 h and in turtles fed a high-acid load for 24-48 h. Intracellular pH was lower in bladders from turtles fed a high-acid load for 48 h but it was not different from controls in the other groups, indicating that intracellular pH cannot account for the adaptive increase in H+ secretion. Bladders from all groups fed an acid load had a higher number of cells with positive staining for acridine orange compared with controls. Double labeling with acridine orange and the mitochondrial stain rhodamine 123 or 6-carboxyfluorescein showed a significant increase in the number of mitochondria-rich cells between control and bladders from turtles fed an acid load. The increase in the number of rhodamine 123- or 6 carboxyfluorescein-positive cells was lower than the increase in acridine orange positive cells, suggesting that the apparent increase in the number of acridine orange-positive cells is due to an increase in the number of acidic vesicles in the mitochondria-rich cells and in the granular cells rather than solely to an increase in the number of mitochondria-rich cells. Plasma membrane fraction prepared from control and acidotic bladders failed to disclose an increase in the putative H+-ATPase as assessed by enzymatic activity and transport studies. In conclusion, the present study suggests that the adaptive increase in H+ secretion in metabolic acidosis is associated both with an increase in the number of mitochondria-rich cells as well as with an increase in the number of acidic vesicles in these cells. PMID- 3028171 TI - Two renal alpha 2-adrenergic receptor sites revealed by p-aminoclonidine binding. AB - [3H]p-aminoclonidine [3H]PAC, a specific alpha 2-adrenergic agonist, was used to characterize alpha 2-adrenoceptor binding in rat renal membranes. Rosenthal plots demonstrated two binding sites with Kds of approximately 1.7 and 14.2 nM and Bmaxs (maximum binding) of 47.3 and 218.8 fmol/mg protein for the high- and low affinity sites, respectively. These characteristics were confirmed by estimate of Kd parameters based on association and dissociation experiments. Pseudo-Hill coefficients generated from drug inhibition experiments were all less than unity, suggesting differential binding at two alpha 2-adrenoceptor binding sites. Specific alpha 2-adrenergic agonists exhibited greater binding affinity to both sites than did nonspecific drugs, and all drugs displayed greater affinity for the high- than the low-affinity binding site. Both guanyl nucleotides and sodium chloride inhibited [3H]PAC binding more at the high-affinity than at the low affinity site. Renal denervation resulted in significant upregulation of receptor density only at the high-affinity sites with no change in receptor affinity at either site, suggesting that a majority of the alpha 2-adrenoceptors in the kidney are postsynaptic. Thus all lines of evidence in this study indicate that two alpha 2-adrenoceptor binding sites exist in the rat kidney. PMID- 3028173 TI - Lack of effect of atriopeptin II on rabbit glomerular arterioles in vitro. AB - This study was designed to determine if atriopeptin II (AP II) has any direct action on glomerular arterioles in vitro. Single superficial afferent or efferent arterioles were dissected from rabbit kidney, cannulated with micropipettes, and changes in lumen diameter were measured. The effect of AP II (10-(12)-10(-7) M) was tested alone or in arterioles precontracted with norepinephrine (3 X 10(-7) M, afferent) or angiotensin II (10(-10) M, efferent). By itself, AP II had no effect on lumen diameter in afferent or efferent arterioles, but both arterioles responded to 3 X 10(-7) M norepinephrine with 74 and 54% reductions in lumen diameter, respectively. Similarly, in norepinephrine-contracted afferent arterioles and angiotensin II-contracted efferent arterioles, AP II did not alter lumen diameter although acetylcholine (10(-6) M) produced near maximal relaxation of both arterioles. Furthermore, luminal addition of AP II (10(-7) M) had no effect on control lumen diameter or on the response to norepinephrine or angiotensin II. The preparation of AP II was biologically active since it caused dose-dependent increases in guanosine 3',5'-cyclic monophosphate levels in isolated rat glomeruli. These results suggest that the renal vascular effects of AP II observed in vivo are not due to a direct action on superficial glomerular arterioles but rather to an, as yet, unidentified indirect action of this peptide. PMID- 3028172 TI - Renal effects of selective adenosine receptor agonists in anesthetized rats. AB - Exogenous adenosine affects renal hemodynamics, renal tubular transport processes, and the secretion of renin. However, adenosine is not a selective agonist; it activates both A1 and A2 cell-surface receptors and it binds to an intracellular P-site that inhibits adenylate cyclase activity. Recent in vitro studies have suggested that activation of A1- and A2- adenosine receptors results in opposite effects on renin secretion. The purpose of these experiments was to examine the renal effects of A1- and A2-adenosine receptor agonists in vivo. 5'-N ethylcarboxamide adenosine (NECA), 2-chloroadenosine (2-CLA), and N6 cyclohexyladenosine (CHA) were infused intravenously at rates that produced comparable decreases in systemic arterial blood pressure. All three of these adenosine analogues produced comparable decreases in para-aminohippurate (PAH) and inulin clearances and in Na and K excretion rates. CHA, an A1-selective agonist, markedly decreased plasma renin concentration (PRC), whereas NECA, an A2 selective agonist, markedly increased PRC; 2-CLA, a nonselective agonist, produced a smaller increase in PRC. Taken together, these results suggest that occupation of A1- and A2-receptors inhibits and stimulates renin secretion in vivo, independently of the effects of these adenosine receptor agonists on arterial blood pressure, renal hemodynamics, and tubular Na and K transport. PMID- 3028174 TI - Evidence that parallel Na+-H+ and Cl(-)-HCO3-(OH-) antiporters transport NaCl in the proximal tubule. AB - The present in vitro microperfusion study examined whether active NaCl transport in the proximal convoluted tubule (PCT) occurs via parallel Na+-H+ and Cl(-)-HCO3 (OH-) exchangers. PCT were perfused with a high-chloride, low-bicarbonate solution simulating late proximal tubular fluid, and were bathed in a similar solution containing 6 g/dl albumin. In this setting the driving forces responsible for passive NaCl transport are eliminated. Addition of 0.1 or 0.5 mM luminal 4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonic acid (SITS), 0.5 mM luminal 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS), or 0.1 mM bath ethoxyzolamide, a lipophilic carbonic anhydrase inhibitor, resulted in an approximately 50% reduction in volume absorption. Inhibition of the Na+-H+ antiporter with 1.0 mM luminal amiloride inhibited volume absorption by 50%. The transepithelial potential difference (PD) was not significantly different from zero, consistent with an electroneutral mechanism for active NaCl transport. The effect of a Cl(-)-HCO3-(OH-) exchanger on acidification was examined in PCT perfused with an ultrafiltrate-like solution and bathed in a serumlike albumin solution. Addition of 0.5 mM DIDS did not significantly decrease volume absorption, demonstrating that luminal DIDS did not result in a nonspecific decrease in solute transport. Luminal DIDS significantly stimulated bicarbonate absorption, consistent with a Na+-H+ antiporter running in parallel with a Cl(-) HCO3-(OH-) antiporter, which exchanges luminal Cl- for cellular HCO3- (or OH-). In conclusion, these data are consistent with parallel Na+-H+ and Cl(-)-HCO3-(OH ) antiporters mediating neutral active NaCl transport in the PCT. PMID- 3028175 TI - Comparison of effects of forskolin, cAMP, and vasopressin on Pf/Pd(w) of toad urinary bladder luminal membrane. AB - We have reported that Pf/Pd(w) (the ratio of osmotic and diffusional water permeabilities) for the luminal membrane of toad urinary bladder is approximately 17 for tissues stimulated with either vasopressin or 8-bromoadenosine 3',5' cyclic monophosphate (8-BrcAMP). In a recent abstract, Kachadorian and co-workers have shown that tissues stimulated with adenosine 3',5'-cyclic monophosphate (cAMP) or forskolin have a lower Pf than would be anticipated from the frequency of aggregates visualized on the flat portion of the luminal membrane using freeze fracture electron microscopy. We report here measurements of Pf/Pd(w) for the luminal membrane of tissues receiving these agents: Pf/Pd(w) for submaximally stimulated tissues was the same, regardless of whether the stimulant was vasopressin (12.7 +/- 0.3), forskolin (13.7 +/- 0.9), or cAMP (12.0 +/- 1.3). The calculated Pd(w)'s for the series barrier were also identical (6.8 +/- 0.5, 6.5 +/- 0.3, and 8.2 +/- 1.0 X 10(-4) cm/s respectively). Our data, taken together with those of Kachadorian et al. are consistent with a number of possibilities: because our methodology does not permit estimation of Pf for the series barrier, we cannot rule out the possibility of a "post-luminal barrier" that is rate limiting for Pf, but not for Pd(w) in forskolin- and cAMP-stimulated tissues, Pf and Pd(w) of the luminal surface aggregates could decrease in parallel, so that luminal membrane Pf/Pd(w) remains constant, and there could be a diminished frequency of fused aggregate-rich aggrephores, but not of aggregates that are on the flat portion of the luminal membrane. Only the latter can be unequivocally quantitated using freeze-fracture electron microscopy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3028176 TI - O2 free radicals: cause of ischemia-reperfusion injury to cardiac Na+-K+-ATPase. AB - The role of O2 free radicals in the reduction of sarcolemmal Na+-K+-ATPase, which occurs during reperfusion of ischemic heart, was examined in isolated guinea pig heart using exogenous scavengers of O2 radicals and an inhibitor of xanthine oxidase. Ischemia and reperfusion reduced Na+-K+-ATPase activity and specific [3H]ouabain binding to the enzyme in ventricular muscle homogenates and also markedly lowered sodium pump activity estimated from ouabain-sensitive 86Rb+ uptake by ventricular muscle slices. These effects of ischemia and reperfusion were prevented to various degrees by O2-radical scavengers, such as superoxide dismutase, catalase, dimethyl-sulfoxide, histidine, or vitamin E or by the xanthine oxidase inhibitor, allopurinol. The degree of protection afforded by these agents paralleled that of reduction in enhanced lipid peroxidation of myocardial tissue as estimated from malondialdehyde production. These results strongly suggest that O2 radicals play a crucial role in the injury to sarcolemmal Na+-K+-ATPase during reperfusion of ischemic heart. PMID- 3028177 TI - Increased myocardial beta-receptors and adrenergic responses in hyperthyroid pigs. AB - Controversy exists presently as to whether thyroid hormone potentiates the action of catecholamines on the heart. Therefore, the relationships between adrenergic sensitivity, myocardial beta-receptor number, and the cardiovascular responses associated with excess thyroid hormone were investigated in pigs (Sus scrofa). A hyperthyroid state was induced by the administration of triiodothyronine (T3; 1 mg/kg iv). After 7 days there was a significant increase in resting heart rate, systolic blood pressure, rate-pressure product, and O2 consumption in the hyperthyroid state. At this time echocardiography showed a substantial increase in myocardial cross-sectional size. Pharmacological tests showed an increased intrinsic heart rate (127 +/- 29 to 205 +/- 25 beats/min; P less than 0.001) and an increased chronotropic sensitivity to isoproterenol. The concentration of isoproterenol required for a 50% of maximal response (ED50) was reduced by 33 +/- 30% (2.1 +/- 1.0 to 1.2 +/- 0.3 micrograms/l; P less than 0.025). The slope of the line relating isoproterenol concentration and change in heart rate was increased by 29 +/- 33% (61 +/- 10 to 78 +/- 10; P less than 0.025). Radioligand studies demonstrated an increase in the number of beta-receptors in right atrial membranes from hyperthyroid animals (41 +/- 7 vs. 75 +/- 18 fmol/mg; P less than 0.02). The apparent dissociation constant (KD) of the receptor for l isoproterenol was similar in membranes from euthyroid and hyperthyroid animals (157 +/- 57 vs. 219 +/- 59 nM, respectively; P = NS). This study demonstrates that hyperthyroidism is associated with an increased chronotropic sensitivity to isoproterenol, consequent to an up-regulation of beta-adrenergic receptors in the right atrium. PMID- 3028178 TI - Beta-adrenergic function in a congestive cardiomyopathy model. AB - We investigated the cardiac beta-adrenergic-adenylate cyclase (AC) system in turkeys inbred for congestive cardiomyopathy (CCM) and the relation of this system to echocardiographically determined left ventricular shortening fraction (LVSF) in individual 56-day-old birds. The isoproterenol (Iso)-stimulated, NaF stimulated, and basal AC activities in CCM birds were decreased to 69, 72, and 86%, respectively, of control values, P less than 0.05. By linear regression analysis there was significant direct correlation of initial LVSF in CCM birds with each of the following: change in heart rate (HR) after Iso function; Iso stimulated, NaF-stimulated, and basal AC activities, and density of beta adrenergic receptors. In addition, we investigated the effect of Iso infusion (0.4 microgram X kg-1 X min-1) on systemic hemodynamics. In CCM birds the initial LVSF was 72% of the control value, P less than 0.01; the initial HR was 112% of the control value, P less than 0.01. During Iso infusion both the LVSF and HR in control birds were increased, whereas in CCM turkeys with marked cardiac dilatation neither of these parameters changed. In 10-day-old CCM turkeys, which show no cardiac dilatation, NaF-stimulated AC activity was reduced to 70% of the control values, whereas basal and Iso-stimulated AC activities were unchanged. Thus an abnormality in the beta-adrenergic-adenylate cyclase system is present in CCM birds before development of cardiac dilatation; when cardiac dilatation is severe, a global defect in this system can be directly related to depressed inotropic and chronotropic responsiveness to Iso infusion in individual turkeys. PMID- 3028179 TI - Myocardial adrenergic responsiveness after lethal and nonlethal doses of endotoxin. AB - A dose-dependent impairment of intrinsic myocardial performance has been observed following in vivo administration of endotoxin. The present study reports a dose dependent increase in plasma catecholamines following endotoxin (ET) that may impair beta-adrenergic responsiveness. Hearts were removed from pentobarbital anesthetized rats 4 h after a bolus injection of saline or ET (1,000, 100, or 10 micrograms/100 g body wt) and were studied as isolated cell preparations following collagenase digestion. Responsiveness of isoproterenol-stimulated adenosine 3',5'-cyclic monophosphate (cAMP) accumulation in myocytes prepared from hearts of animals injected with 10 and 100 micrograms ET was decreased when compared with control rats and was significantly blunted in myocytes prepared from animals receiving 1,000 micrograms ET. Similar sensitivities of the cAMP system existed, as judged by similar half-maximum effective concentration values. cAMP accumulation in the presence of 1 microM forskolin was depressed in myocytes from the 1,000-microgram ET animals; beta-adrenergic receptor density was decreased 25% (P less than 0.05) in myocytes from high-dose ET animals when compared with control animals. This was accompanied by a nonsignificant reduction in the affinity of binding sites for (+/-) [3H]CGP 12177. The blunted myocyte hormonal responsiveness following ET challenge appears to be related to the decreased activity of the adenylate cyclase that may be attributed to alterations in both receptor density and in the adenylate cyclase itself. PMID- 3028180 TI - Cardiac muscle ultrastructure and cyclic AMP reactions to altered gravity conditions. AB - Morphological and biochemical analyses of heart muscle of rats subjected to microgravity on Spacelab 3 (SL-3) flight and rats born and reared under increased gravity (1.7 G) conditions were compared with 1-G controls. Electronmicroscopic studies showed an increase in the number of lipid droplets and in areas of glycogen storage. Distribution changes of microtubules and cytoskeletal elements from both SL-3 and 1.7-G groups were observed. The high Km cyclic AMP phosphodiesterase activity was lower (P less than 0.05) in SL-3 heart muscle, and low Km activity was lower in 1.7-G males but was unaltered in females. Cyclic AMP dependent protein kinase (cA-PK) activity was decreased in subcellular fractions of heart muscle of SL-3 animals. Recompartmentalization of cA-PK activity occurred in particulate tissue fraction of 1.7-G animals (70.3% of total for 1.7 G vs. 35.9% for controls). Phosphorylation of endogenous low-mobility proteins increased in SL-3 heart-soluble fractions. Photoaffinity labeling (18 h, 4 degrees C) decreased in type II cA-PK regulatory (R) subunits in both SL-3 and in 1.7-G male heart tissue particulate fractions. The 1.7-G female heart R subunit distribution did not differ from controls. These findings indicate that in heart muscle altered gravity conditions influenced physiological reactions similar to catecholamine-induced receptor-mediated hormonal responses. PMID- 3028181 TI - Salivary gland ultrastructure and cyclic AMP-dependent reactions in Spacelab 3 rats. AB - Environmental stimuli influencing catecholamine levels induce changes in cyclic AMP-dependent reactions and cell morphology in the rat parotid. Responses of salivary glands to spaceflight were determined by measurement of cyclic AMP mediated reactions in fresh-frozen salivary glands and by microscopic evaluation of ultrastructure in fixed parotid glands. Decreased cell-free protein phosphorylation occurred in parotid glands in three of five flight animals. Protein kinase activity ratios were decreased in the soluble and increased in the particulate fractions of Spacelab 3 (SL-3) rat sublingual glands, compared with ground controls. Biochemical analyses show that effects of space flight on salivary glands are similar to those induced experimentally by physiological manipulation or alteration of catecholamine levels. Morphological evaluation of three SL-3 rat parotid glands showed increased numbers of lysosomes, autophagic vacuoles containing degenerating secretory product, and accumulation of lipid droplets. Since these animals lost weight, consistent with disruption of food and water consumption, morphological changes may in part be due to decreased masticatory stimulation, as occurs with reduced food intake or a liquid diet. The observed changes may reflect physiological responses of the gastrointestinal and autonomic systems to effects of spaceflight. PMID- 3028182 TI - Role of adrenergic-dependent H+ release from red cells in acidosis induced by hypoxia in trout. AB - The response to severe hypoxia is characterized in trout by a sudden drop in blood pH, which is of metabolic origin, and by an increase in the blood concentration of adrenaline. This acidification is biphasic in nature. The first phase of acidification is not associated with a rise in the blood lactate concentration and no longer occurs after pretreatment of the fish with a beta blocker agent, propranolol. Thus an acid other than lactic acid is released into the blood at the onset of hypoxia and this release, which is under beta adrenergic control, is responsible for the first phase of acidification. On the other hand the second phase of acidification is related to an increase in blood lactate and is not modified by a beta-blocker agent. We have also demonstrated that deep hypoxia promotes a rapid increase in red blood cell volume and that this cell enlargement is coincident with a large net uptake of Na+ and Cl-. In the presence of beta-blocking agents the Na+ uptake is blocked and the swelling of the cells is considerably inhibited. The residual swelling is clearly due to the chloride shift induced by both deoxygenation of hemoglobin and change in blood pH. In the light of data obtained in vitro on the effect of catecholamines on trout erythrocytes, it can be considered that the first phase of acidification occurring at the onset of hypoxia, and that is under beta-adrenergic control, is due essentially to the release of H+ by red blood cells in exchange with external sodium mediated by a beta-adrenergic-stimulated Na+-H+ exchanger.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3028183 TI - Responses to converting-enzyme inhibition and hemorrhage in newborn lambs and adult sheep. AB - We compared the cardiovascular and hormonal responses to angiotensin converting enzyme inhibition and hemorrhage of 20% of blood volume in chronically instrumented unanesthetized newborn lambs and adult sheep. Administration of the nonsulfhydryl-containing converting-enzyme inhibitor enalapril reduced mean arterial pressure in the newborn but not in the adult animals. Blood pressure fell in both age groups after hemorrhage, and the hemorrhage-induced fall in blood pressure, integrated over the period of hypovolemia, was more pronounced when converting-enzyme inhibition was present in the lambs. This was not observed in the adults. Cardiac output fell following hemorrhage in both age groups, and the fall was greater when enalapril was present in the lambs, but this was not the case in the adults. Hemorrhage increased plasma renin activity in both groups, and enalapril augmented this increase. Plasma concentrations of vasopressin and catecholamines increased following hemorrhage within and between groups. Taken together these data suggest that the renin-angiotensin system plays a more important role in the maintenance of cardiovascular homeostasis in newborn lambs than it does in adult sheep, and catecholamine and vasopressin responses to volume loss can occur in the presence of blockade of the renin-angiotensin system. PMID- 3028184 TI - Secretory activity in salt glands of birds and turtles: stimulation via cyclic AMP. AB - O2 consumption in tissue slices from the nasal salt gland of the duck and the lachrymal salt gland of Malaclemys is stimulated by methacholine, a stimulation that is inhibited by bumetanide and by ouabain. In addition, the calcium ionophore A23187 mimics the action of methacholine in stimulating this secretion related O2 consumption in both glands, suggesting a second-messenger role for this ion in the cholinergic response. However, the adenylate cyclase activator, forskolin, and the cyclic AMP analogue, 8-cpt-cAMP, also stimulate ouabain sensitive and bumetanide-sensitive O2 consumption in both the duck gland and the Malaclemys gland. It is suggested that the mechanism of salt secretion in the Malaclemys lachrymal gland conforms to that previously described for other extrarenal salt-secreting tissues in nonmammalian vertebrates and, as in the bird gland, is subject to a cholinergic regulation potentially acting via changes in intracellular calcium. In addition to this, secretory activity in both the avian and the turtle glands can be stimulated by a previously undisclosed adenylate cyclase-cyclic AMP system. The identity of the primary signal for such a system is not known, nor is the nature of any interrelationship between the two second messenger systems that have been identified in these glands. PMID- 3028185 TI - Blunted aldosterone and ACTH release after human CRH administration in depressed patients. AB - The authors measured pituitary adrenocortical responses to human corticotropin releasing hormone (CRH) in 10 patients with a major depressive episode and 10 matched control subjects. Depressed patients had a significantly lower aldosterone and ACTH release, but cortisol and corticosterone responses were not different between groups. PMID- 3028186 TI - Carry-over effects of marijuana. PMID- 3028187 TI - Norwalk-like gastroenteritis epidemic in a Toronto hospital. AB - An epidemic of gastroenteritis in a teaching hospital affected 57 patients and 69 staff over a 26-day period. The index case was a patient admitted with acute abdominal pain and diarrhea two days prior to the outbreak. The epidemic curve indicated person-to-person transmission. The incubation period, duration and types of symptoms were typical of Norwalk gastroenteritis, and Norwalk-like virus particles, serologically different from the prototype Norwalk virus strain, were observed in 17 of 20 fecal specimens examined by immune-electron microscopy. PMID- 3028188 TI - Immunity to cytomegalovirus in women with unexplained recurrent spontaneous abortion. AB - Humoral and cellular immunity to cytomegalovirus (CMV) has been assessed in women suffering unexplained recurrent spontaneous abortions (RSA). A significantly lower prevalence of serum anti-CMV antibodies was observed for RSA women compared with either their male partners or age-matched female controls, unlike for serum antibodies to Herpes simplex virus. In addition, there was a markedly impaired lymphocyte proliferative response to CMV for CMV-sero-positive RSA women compared with CMV-seropositive controls. These results indicate that women with unexplained RSA have difficulty in responding to CMV, and are of significance when considering leukocyte transfusion immunotherapy. PMID- 3028189 TI - Surgical pathology of the infected gut. AB - In the normal digestive tract, interaction of the mucosa with a large and varied microbial flora is inevitable. In fact, the "normal" state of the digestive tract reflects the impact of the resident flora to a significant degree. The pathogenesis of various infectious conditions encountered by the surgical pathologist in the gut is understood more readily when the gastrointestinal tract is viewed as a complex ecosystem. Significant disease may result from perturbation of the normal flora, as well as from exogenous infection, and susceptibility of the host may vary with disturbances of the digestive ecosystem. An ecologic perspective is essential in the consideration not only of the "infected" gut, but of conditions as diverse as ischemia or even carcinogenesis in the gastrointestinal mucosa. PMID- 3028190 TI - Tissue diagnosis of selected AIDS-related opportunistic infections. AB - Opportunistic infections are the most common initial manifestations of AIDS and, in many instances, are first encountered in surgical specimens. Pneumocystis carinii pneumonitis is by far the most frequent infection seen in biopsy specimens of AIDS patients. Most pathologists are familiar with its histopathologic presentations from previous experience. By contrast, many other opportunistic infections are either new or present clinically and pathologically in unfamiliar ways. Cytomegalovirus affects primarily the alimentary tract and lung. Colitis is the most common presentation. Penetrating ulcers may perforate. Most often, mesenchymal cells, endothelium in particular, show the typical intranuclear and intracytoplasmic inclusion bodies. The greater the number of inclusion bodies in tissues the shorter is the survival of the patient. Mycobacterium avium-intracellulare affects mainly small intestine and lymph nodes and produces a clinical and histologic picture similar to that of Whipple's disease. Diffuse infiltrates of histiocytes stuffed with acid-fast bacilli are characteristic. Cryptosporidiosis is the most ominous enteric opportunistic infection. Protozoa attach themselves to the epithelial surface and produce severe profuse, watery diarrhea. Cryptococcosis is seen in lung, lymph node, and brain biopsy specimens. Large numbers of organisms, sometimes with deficient mucinous capsules, and little or no inflammatory reaction, are notable. Toxoplasmosis is the most common cause of neurological complications in AIDS. Brain biopsy specimens show necrosis, microglial nodules, perivascular lymphocytic infiltrates, and, in 50% of cases, trophozoites. PMID- 3028191 TI - Epizootic vesicular stomatitis in Colorado, 1982: epidemiologic and entomologic studies. AB - An epizootic of vesicular stomatitis (VS) caused by the New Jersey serotype of VS virus affected livestock and humans in 14 western states in 1982-1983. Epidemiological observations were made on at least 10% of the cattle in 4 dairy herds that were located in the vicinity of Grand Junction, Colorado. High rates of neutralizing antibody to the New Jersey serotype were seen in all cattle regardless of whether livestock in the dairy had clinical VS or a decrease in mild production. Antibody titers remained high in these cattle for as long as 2 years after the epizootic. No virus isolations were made from 32 humans with clinical signs compatible with viral disease. Entomological information was obtained during the epizootic from 23 premises in northwestern Colorado. Insect collections yielded 4 isolates from Culicoides spp. midges, 2 from C. variipennis, and 1 each from C. stellifer and C. (Selfia) spp. This is the first report of VS virus isolations from field-collected Culicoides. PMID- 3028192 TI - Epizootic vesicular stomatitis in Colorado, 1982: infection in occupational risk groups. AB - In 1982-1983, an epizootic of vesicular stomatitis occurred in the western United States. Veterinarians, research workers, and regulatory personnel who were exposed to vesicular stomatitis virus were examined for patterns of human infection and prevalence of vesicular stomatitis New Jersey serotype neutralizing antibody. Insight into the mechanism of transmission was sought by comparing activities of antibody-positive and antibody-negative persons. A statistically significant risk factor was a history of infected animals sneezing in the face of serosurvey participants. Elevated odds ratios were also calculated for those who usually examined the oral cavity of affected animals, had open wounds on hands or arms, and had exposure to saliva through the eye or skin. Relatively intimate direct contact was required; a higher risk was associated with examining horses than cattle. Neutralizing antibody prevalence was significantly higher among exposed persons with illness (23%) than in exposed persons without a history of clinical illness (7%). Overall, however, infectivity of VSNJ for humans during the epizootic was low. PMID- 3028194 TI - Maintenance of Toscana virus in Phlebotomus perniciosus by vertical transmission. AB - Toscana virus was maintained in a laboratory colony of Phlebotomus perniciosus by vertical (transovarial) transmission for 13 consecutive generations over a 23 month period. No significant biological changes were noted in the virus after prolonged vertical passage in the sand flies, and transovarially infected females were able to transmit the agent by bite to susceptible animals. Chronic infection of Ph. perniciosus with Toscana virus had no apparent effect on the insects' rate of eclosion. In the absence of selection and with random matings, the virus infection rates in each subsequent generation of the colony decreased, suggesting that Toscana virus cannot be maintained in Ph. perniciosus by transovarial transmission alone. Alternative mechanisms for virus maintenance are discussed. PMID- 3028193 TI - Epizootic vesicular stomatitis in Colorado, 1982: epidemiologic studies along the northern Colorado front range. AB - Epidemiologic evaluations were made of farm personnel on vesicular stomatitis affected premises along the front range of the Rocky Mountains in Colorado during the 1982 epizootic. A similar antibody prevalence was noted to that of veterinarians and research and regulatory personnel who were involved with the same epizootic. Risk of infection resulted from intimate physical contact with infected horses or cows. Incidence and infection rates in horses were 45%; rates in cows were much lower, only 5%. Some epidemiologic clues were gained by a detailed study of an equine ranch. The pasture was incriminated as the area of highest risk, where 100% infection rates were noted. Horses in open pens and barns were at lower risk. Severe clinical disease in horses resulted in higher neutralizing antibody titers than inapparent or mild infection. Maternal antibody was detected in foals up to 4 months of age, and the level of antibody in the foal was a reflection of the dam's antibody level. PMID- 3028195 TI - [Effect of GABA-linoleamide and glycine-linoleamide on catalepsy in the rat after chronic administration of reserpine]. PMID- 3028196 TI - [Combined chemotherapy of uterine trophoblastic tumors]. PMID- 3028197 TI - Atypical cutaneous fibrous histiocytoma. AB - We report seven cases of atypical cutaneous fibrous histiocytoma, which appears to be a variant of cutaneous fibrous histiocytoma (dermatofibroma). These patients are all middle-aged women (mean, 39 years old) with small nodules occurring on the trunk and limbs. The lesions are characterized by marked focal cellular atypia, the absence of mitoses, and xanthomatous changes in both mononuclear and giant cells--all found within a small dermal nodule (approximately 1 cm) separated from an acanthotic epidermis by a Grenz zone. Their benign nature is demonstrated by the absence of recurrence, even after 9 years follow-up time. PMID- 3028198 TI - The Paget cell. Mistaken for a parasite in the 19th century. AB - Sir James Paget in 1874 described clinically a disease of the skin that bears his name. Its dermatopathologic aspect was partly elucidated during the years that followed. This led to a theory that the Paget cell was a carcinogenic parasitic body. George Thomas Beatson, on recognizing the erroneous nature of the parasitic theory of cancer, was led logically to propose and successfully to carry out bilateral ovariectomy for advanced cancer of the breast by 1896. PMID- 3028199 TI - Ethanol-induced inhibition of mouse brain adenosine triphosphatase activities: lack of interaction with norepinephrine in vitro. AB - The in vitro effects of ethanol on C57BL/6J mouse forebrain ATPase activities were investigated in the presence and absence of norepinephrine. Three enzyme activities (Na + K-ATPase, Mg-ATPase, and low affinity Ca-ATPase) were studied in forebrain homogenates using a colorimetric assay. The effect of norepinephrine on ethanol inhibition of Sprague-Dawley rat brain Na + K-ATPase activity was examined in selected experiments for direct comparison with the results obtained using mouse brain. Ethanol (250-2000 mM) inhibited all ATPase activities in a concentration-dependent manner. In each case the IC50 was well beyond what would be a lethal concentration in vivo. Ethanol inhibition of mouse forebrain Na + K ATPase activity was competitive with regards to K+ ion concentration, but was uncompetitive for inhibition of Mg-ATPase and Ca-ATPase activities. Norepinephrine (0.1 mM) did not sensitize mouse brain Na + K-ATPase activity to ethanol-induced inhibition. In contrast, norepinephrine sensitized rat brain Na + K-ATPase to ethanol inhibition when tested simultaneously with mouse brain under identical conditions. These results cannot be explained by differences in assay conditions and suggest that the interaction between norepinephrine and ethanol inhibition of Na + K-ATPase activity may be species specific. Norepinephrine alone stimulated mouse and rat brain Na + K-ATPase activity when assayed in imidazole buffer, but not when assayed in tris buffer. Furthermore, 0.1 mM norepinephrine slightly antagonized the inhibitory effect of ethanol on mouse brain Mg-ATPase activity, but did not affect ethanol-induced inhibition of Ca ATPase activity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3028200 TI - Cell receptors and ethanol. PMID- 3028201 TI - [Cystic adenoid carcinoma of the upper lip]. PMID- 3028203 TI - Consecutive DNA terminator sequencing by using enzymatically generated primers. AB - An improved strategy for the dideoxy chain termination sequencing of M13 recombinant DNA using enzymatically generated primers is presented. It involves synchronous extension of the universal primer with the Klenow fragment of DNA polymerase I at an average rate of 200 deoxynucleoside triphosphates per minute to the immediate downstream region of a preselected restriction enzyme site. Subsequent cleavage with the restriction enzyme generates the short primers with homogeneous 5' ends which can be used for further sequencing. The next restriction sites are selected in the newly sequenced regions of DNA by means of a microcomputer. By repeating this primer extension-cleavage-sequencing strategy sequences altogether about 6 kb long from several recombinant single-stranded M13 DNAs have been determined by using twenty restriction enzymes with different specificities. PMID- 3028202 TI - Combination treatment with ranitidine and sodium bicarbonate prior to obstetric anaesthesia. AB - The gastric pH and volume were measured in 175 patients undergoing elective, and 313 undergoing emergency, obstetric procedures. Ranitidine 150 mg was administered orally every 6 hours in labour and at least 2 hours before elective Caesarean section. Patients received 20 ml of 8.4% sodium bicarbonate orally immediately prior to induction of anaesthesia. The combination of ranitidine and sodium bicarbonate produced marked alkalinisation of gastric contents (mean pH 8.9). The administration of sodium bicarbonate pre-operatively in patients who received ranitidine less than 2 hours before operation led to satisfactory elevation of gastric pH. Only four patients had a gastric pH less than 2.5, one patient refused any medication, two received only ranitidine and one patient had a long interval from administration of bicarbonate to aspiration of gastric contents. Gastric volumes were high in labouring patients (mean 84 ml) despite administration of ranitidine. The effectiveness of sodium bicarbonate as a single dose antacid therapy prior to obstetric anaesthesia requires further study. PMID- 3028204 TI - T4 polynucleotide kinase: macromolecular crowding increases the efficiency of reaction at DNA termini. AB - The amount of reaction catalyzed by T4 polynucleotide kinase on a variety of its substrates is greatly increased in the presence of polyethylene glycol 8000 (PEG 8000). Both the forward and reverse reactions as well as the exchange reaction can be stimulated. The stimulation is a general effect on T4 polynucleotide kinase reactions involving high molecular weight DNA substrates. The use of PEG 8000 is particularly advantageous for labeling or removing terminal 5'-phosphate groups which are only slowly or incompletely labeled or removed under ordinary conditions, such as those at recessed termini or at "nicks" in duplex DNA, although the reaction on blunt-ended or protruding termini is also increased. It is further advantageous for labeling very low concentrations of substrates. PMID- 3028206 TI - Accurate, quantitative assays for the hydrolysis of soluble type I, II, and III 3H-acetylated collagens by bacterial and tissue collagenases. AB - Accurate and quantitative assays for the hydrolysis of soluble 3H-acetylated rat tendon type I, bovine cartilage type II, and human amnion type III collagens by both bacterial and tissue collagenases have been developed. The assays are carried out at any temperature in the 1-30 degrees C range in a single reaction tube and the progress of the reaction is monitored by withdrawing aliquots as a function of time, quenching with 1,10-phenanthroline, and quantitation of the concentration of hydrolysis fragments. The latter is achieved by selective denaturation of these fragments by incubation under conditions described in the previous paper of this issue. The assays give percentages of hydrolysis of all three collagen types by neutrophil collagenase that agree well with the results of gel electrophoresis experiments. The initial rates of hydrolysis of all three collagens are proportional to the concentration of both neutrophil or Clostridial collagenases over a 10-fold range of enzyme concentrations. All three assays can be carried out at collagen concentrations. that range from 0.06 to 2 mg/ml and give linear double reciprocal plots for both tissue and bacterial collagenases that can be used to evaluate the kinetic parameters Km and kcat or Vmax. The assay developed for the hydrolysis of rat type I collagen by neutrophil collagenase is shown to be more sensitive by at least one order of magnitude than comparable assays that use rat type I collagen fibrils or gels as substrate. PMID- 3028205 TI - Properties of radiolabeled type I, II, and III collagens related to their use as substrates in collagenase assays. AB - Calf skin and rat tendon type I, bovine cartilage type II, and human amnion type III collagens have been radiolabeled by reaction with [3H]acetic anhydride, [3H]formaldehyde, and succinimidyl 2,3-[3H]propionate. All three reactions produce collagens with high specific activities that are suitable for use as substrates in collagenase assays. The identity of the radiolabel and the labeling indices do not alter the molecular weights or thermal stabilities of the collagens or the solubilities of the collagens or gelatins in dioxane-water mixtures at 4 degrees C. However, in contrast to native or sparsely labeled collagens, those with 40 or more lysine + hydroxylysine residues labeled per molecule do not undergo fibrillogenesis in the presence of 0.2-0.4 M NaCl in the 4-35 degree C temperature range. Thus, the modification reactions not only serve to introduce the radiolabel, but also to keep the collagens soluble over a wide range of temperatures and concentrations. The TCA, TCB fragments produced on partial reaction of each collagen type with tissue collagenases can be selectively denatured by a 10-minute incubation under specific conditions and the intact collagens selectively precipitated by addition of 50% v/v dioxane. This serves as the basis for soluble collagenase assays. The effect of labeling index on the properties of the collagens has been investigated and the results establish the range of conditions over which these collagens can be used as substrates for soluble versus fibrillar collagenase assays. PMID- 3028208 TI - Separation of phosphoinositides and other phospholipids by two-dimensional thin layer chromatography. AB - A simple, rapid, two-dimensional TLC system is presented which resolves the four phosphoinositide cycle phospholipids as well as all commonly encountered major and minor phospholipids. Ca2+-free lipid samples are loaded onto silica gel HL plates and developed first in 48:40:7:5 chloroform:methanol:water:concentrated ammonia, and then in 55:25:5 chloroform:methanol:formic acid. The method was applied successfully to human erythrocytes, human platelets, and BL/VL3 murine lymphoma cells. PMID- 3028207 TI - Isolation of cationic peptides from rat polymorphonuclear leukocyte granule contents using fast protein liquid chromatography. AB - Separation of extracted rat polymorphonuclear leukocyte (PMN) granule contents using fast protein liquid chromatography yielded four major protein fractions. These fractions consisted of myeloperoxidase (peak A), neutral protease (peak B), lysozyme (peak C), and low molecular weight, cationic peptides (peak D). This study represents the first noted purification of the cationic peptides of rat PMN granules. PMID- 3028209 TI - A fluorometric method for the determination of functional (Na,K)-ATPase and cardiac glycoside receptors. AB - A method to measure high-affinity binding sites for fluorescent ligands is described. The method is applied to the determination of (Na,K)-ATPase using the fluorescent ouabain derivative anthroylouabain (P. A. G. Fortes (1977) Biochemistry 16, 531-540). The method consists of measurements of the fluorescence intensities of a saturating concentration of anthroylouabain in the presence and absence of (Na,K)-ATPase and ouabain. These data and the fluorescence enhancement factor upon anthroylouabain binding are used to calculate the concentration of binding sites. The measurements can be done in a few minutes and 10 to 100 pmoles of ouabain sites is sufficient for accurate determinations. Because phosphorylation of (Na,K)-ATPase is necessary to bind anthroylouabain, only functional enzyme is detected by this method. PMID- 3028210 TI - A one-step technique for the subcellular fractionation of total cell homogenates. AB - A procedure was developed for the rapid, analytical subcellular fractionation of entire homogenates from the Chinese hamster ovary and HeLa cell lines. The procedure avoids a nuclear sedimentation step and the losses that accompany such a step. A key to the development of this procedure was the addition to homogenates of either micrococcal nuclease or DNase I. Nuclease-treated homogenates were fractionated on self-forming Percoll gradients. The entire procedure from cell harvesting through collecting gradient fractions took only 2.5 h. The position of marker enzymes in the gradient fractions indicated clear resolution of plasma membranes, Golgi apparatus, endoplasmic reticulum, and lysosomes. This procedure should facilitate many studies requiring subcellular fractionation of cultured cells. PMID- 3028211 TI - The preparation of tritiated tunicamycin. AB - A relatively simple and inexpensive procedure was devised for the radiolabeling of the glycoprotein biosynthesis inhibitor, tunicamycin. The procedure is based on hydrogen exchange in alkaline solutions of tritiated water. It was noted that the antibiotic was much more alkali labile than model compounds such as uridine. The alkali stability of the inhibitor was studied to determine conditions for optimum labeling and yield. The effects of alkaline incubation on the inhibitory properties of the antibiotic were also investigated and it was found that the breakdown products are not effective inhibitors of the reaction that transfers N acetylglucosamine-1-phosphate to dolichyl phosphate. The isolated radioactive tunicamycin homologs, however, retained all their inhibitory action. Incubation of tunicamycin in the presence of deuterated water and mass spectral analysis showed that under the conditions used for the tritiation of tunicamycin the major product exchanged six hydrogen atoms. The position of the tritium atoms in labeled tunicamycin was not determined. The radioactive label in these compounds was shown to be stable under physiological conditions and should be useful for investigations involving the action of these antibiotics. PMID- 3028212 TI - Graphic presentation and analysis of inhibition data from ligand-binding experiments. AB - One technique for the characterization of receptor subtypes involves measuring the inhibition of the binding of a radioligand which is not subtype selective by agonists or antagonists which are subtype selective. Although such data are routinely calculated using computer programs, it is often useful to have a graphical representation of the data. Until now, a "modified Scatchard" or "Hofstee" plot of the form P = -(P/I)(IC50) + 1 (where P is percentage inhibition and I is the inhibitor concentration) has been used. We describe an alternate plot of the form B = -(B X I)(1/IC50) + B0 (where B is the concentration of bound radioligand and B0 is the concentration of radioligand bound in the absence of the inhibitor). This method has the important advantage that the data need not be calculated as percentage inhibition which eliminates the error involved in the experimentally determined B0 value being included in the other values. PMID- 3028214 TI - n-Alkyldimethylammonium propanesulfonates as stationary phase modifiers in reversed-phase liquid chromatography. PMID- 3028213 TI - Extraction of adenine nucleotides from cultured endothelial cells. AB - The goal of this work was to find a method suitable for the extraction of adenine nucleotides from cultured vascular endothelial cells. Extraction of cell monolayers with 80% methanol in water yielded extracts with a higher content of ATP than did extraction of cells with perchloric acid, trichloroacetic acid, or boiling water. The optimal extraction solution was 80% methanol with 0.5 mM ethylene glycol bis(beta-aminoethyl ether) N,N'-tetraacetic acid (EGTA) or EDTA, heated to 70 degrees C immediately before use. Extraction of nucleotides by this solution was rapid and the recovery of exogenous ATP added during the extraction process was generally greater than 90%. An aqueous methanol or ethanol solution may be applicable for the extraction of nucleotides and other metabolites from cultured animal cells, dispersed cells, and frozen, powdered tissues. PMID- 3028215 TI - Effect of solvent on solid-supported reactions on styrene/divinylbenzene copolymeric macroreticular resin. PMID- 3028216 TI - Proton nuclear magnetic resonance spectroscopy of aqueous solutions: complete elimination of the water resonance by spin-spin relaxation. PMID- 3028217 TI - A clinical study of sensitivity to sodium nitroprusside during controlled hypotensive anesthesia in young and elderly patients. AB - Aging has important effects on the cardiovascular system; baroreceptor reflex function decreases and the elderly are more resistant to both beta-receptor agonists and antagonists. The purpose of the present clinical study was to determine the relationship between age and sensitivity to sodium nitroprusside in 16 patients during deliberate hypotensive anesthesia by determining the blood pressure changes in young and elderly patients to incremental increases in dose of sodium nitroprusside. A dose-response curve relating change in mean blood pressure to dose of sodium nitroprusside (microgram X kg-1 X min-1) was constructed for each patient; the slope of this line is a measure of "sensitivity." The change in mean arterial blood pressure per microgram X kg-1 X min-1 nitroprusside dose (i.e., slope), showed a significant correlation with age (r = 0.766, P less than 0.001), demonstrating that sensitivity to sodium nitroprusside increases with advancing years. The maximum change in heart rate produced by nitroprusside showed a reciprocal correlation with age (r = -0.791, P less than 0.001). There was no significant correlation between age and maximum change in plasma norepinephrine or epinephrine concentrations during nitroprusside infusion. The increased sensitivity to nitroprusside might have been due to diminished baroreflex activity, resistance of cardiac adrenergic receptors to catecholamine stimulation, or alteration in sensitivity to the direct vasodilating effects of sodium nitroprusside. Whatever the mechanism, however, this clinical study has shown that lower doses of nitroprusside should be used in elderly patients to achieve the same degree of hypotension achieved in younger patients. PMID- 3028218 TI - Bronchoconstrictor effects of leukotriene E4 in normal and asthmatic subjects. AB - The bronchoconstrictor activity of an aerosol of leukotriene E4(LTE4) was compared with that of histamine in 5 normal and in 6 asthmatic subjects to define the relative potency of LTE4 between the groups using 3 indices of airway response. The FEV1 and the flow rate measured at 30% of vital capacity from partial and maximal expiratory maneuvers (V30-P and V30-M) were measured. The geometric mean (GSEM) concentration of LTE4 required to reduce the V30-P by 30% was 0.30 (1.46) mM in the normal subjects, and 0.058 (1.63) in the asthmatic subjects; LTE4 was 39-fold more potent than histamine in the former and 14-fold in the latter group. Further, we observed that when normal and asthmatic subjects were compared at a degree of bronchoconstriction resulting in a 30% decrement in the V30-P after inhaling LTE4, there was a greater response in the asthmatic group than in the normal group of the accompanying change in the FEV1. The decrements in the FEV1 were not significantly different between the 2 groups after inhaling histamine. This study demonstrates that LTE4 is a potent bronchoconstrictor agonist in humans and suggests that airway responsiveness to this agonist differs substantially with the index of bronchoconstriction used for assessment of airway response. PMID- 3028219 TI - An in vitro model for the induction of angiotensin-converting enzyme in sarcoidosis. Evidence for a soluble ACE-inducing factor. AB - Normal peripheral blood T-lymphocytes (T) enhance monocyte (Mo) angiotensin converting enzyme (ACE) production in vitro. To evaluate the possible role of a soluble mediator in enzyme induction, the ability of conditioned medium (CM) from Mo-T cocultures to stimulate ACE in fresh Mo was examined. These studies demonstrated that CM promoted a dose-dependent increase in ACE. Generation of the active factor required both Mo and T during culture since neither cell alone yielded active CM. Time course studies revealed that the ACE-inducing activity appeared in CM after 3 to 4 days of culture and increased steadily thereafter. The CM effectively induced ACE in Mo from several donors in contrast to T dependent induction, which required autologous conditions. The time courses for T dependent and CM-dependent ACE induction were similar, and the results suggested that the Mo must go through some period of maturation before they respond to CM. We conclude that a soluble mediator derived from Mo-T cocultures enhances Mo ACE production. This in vitro system may be a useful experimental model of the in vivo induction of ACE in the Mo-derived sarcoid epithelioid cell. PMID- 3028220 TI - [Congenital mesoblastic nephroma]. AB - Authors report three cases of congenital mesoblastic nephroma. In two of them, diagnosis was neonatal, while the third was made at the 3rd-month. They discuss diagnostic procedures employed as well as pathological findings. Presence of a mass was the first clinical sign in all three cases. Nephrectomy was the only treatment employed, with favourable outcome. PMID- 3028221 TI - Functional renal insufficiency during long-term therapy with captopril and enalapril in severe chronic heart failure. AB - Renal function was evaluated in 104 patients with severe chronic heart failure whom we treated with captopril or enalapril. Seventy patients showed no change or an improvement in renal function (group A), and 34 patients developed functional renal insufficiency (group B). Before converting-enzyme inhibition, group B patients received higher doses of furosemide (p less than 0.02) and had lower central venous pressures (p less than 0.05) than group A patients. After 1 to 3 months of converting-enzyme inhibition, an excessive reduction in left ventricular filling pressure (to less than 15 mm Hg) or mean arterial pressure (to less than 60 mm Hg) was noted in 28 of 34 (82%) patients in group B but in only 22 of 70 patients in group A (31%) (p less than 0.001). At the end of the study, drug-induced azotemia resolved after a reduction in the dosage of diuretics, despite unaltered treatment with captopril and enalapril. Hence, the deterioration of renal function after converting-enzyme inhibition in heart failure is not a toxic or immunologic reaction to therapy but results from specific hemodynamic events that can be ameliorated by sodium repletion. PMID- 3028222 TI - Favorable prognosis of brain metastases in small cell lung cancer. AB - Brain metastases are found at diagnosis in 10% of patients with small cell lung cancer. To clarify the effect of central nervous system metastases on prognosis, the records of 429 patients with small cell lung cancer were reviewed. Forty three patients (10%) presented with brain metastases. In 18 patients the brain was the only site of metastatic disease, whereas the remaining 25 patients had at least one additional metastatic site. Forty-one of forty-three patients were treated with combination chemotherapy and cranial radiotherapy. Systemic response rates were similar for both groups. Twenty-seven patients underwent repeat central nervous system staging: 19 (70%) had a complete response, 4 (15%) a partial response, and 4 (15%) no response. Median survival of patients with only one site of metastatic disease was 11 months; patients with additional sites lived 5 months (p = 0.153). Survival in patients with only one site is similar to that in patients with limited disease (11 compared with 13 months; p = 0.074). PMID- 3028223 TI - [Anatomo-clinical conference. Pitie-Salpetriere Hospital. Recurrent neurological deficits and hypofibrinemia in a 35-year-old man]. PMID- 3028224 TI - [2 cases of AIDS in patients with hemophilia A]. AB - The authors report the first two cases of lethal AIDS in haemophilia A in France. One French case of AIDS in haemophilia B has already been reported. The diagnosis was based on the observation of opportunist infections associated with severe depression of cell-mediated immunity in both cases. The possibility of active transmission of the LAV retrovirus by factor VII concentrate is discussed. This raises the problem of the signification of anti-LAV antibodies in haemophiliac patients and the control of products used for the treatment of haemophilia. PMID- 3028225 TI - [Anatomo-clinical conference. Hopital de la Pitie-Salpetriere. Case no. 5--1986. Amicrobic purulent swelling of the psoas in a young man]. PMID- 3028226 TI - [Epidemiologic study of patients admitted during a 1 year period to psychiatric after care]. PMID- 3028227 TI - Congenital cytomegalovirus infection: prospects for prevention. PMID- 3028228 TI - The diagnostic value of ultrastructural studies of skin-punch biopsies and buffy coat for the early diagnosis of some neurodegenerative diseases. PMID- 3028229 TI - In vitro studies of the blood-brain barrier using isolated brain capillaries and cultured endothelial cells. AB - The endothelial cells in brain capillaries are the anatomic site of the blood brain barrier. To learn more about the biology of these specialized cells, we developed methods to prepare suspensions of purified brain microvessels as well as primary cultures of endothelial cells in monolayer. These two preparations allow for direct investigation of the metabolism, transport properties, and receptor content of the brain capillary. We used isolated brain microvessels to study distribution of membrane carriers between the luminal and the abluminal plasma membrane of endothelial cells. We found that Na+K+-ATPase and the A-system amino-acid transport system are located predominantly on the abluminal surface of brain capillary endothelial cells. This distribution of transport carriers is consistent with the low permeability of potassium and small neutral amino acids in the blood-to-brain direction. It suggests, however, that both solutes can be actively transported across brain capillaries from the brain interstitial fluid to the blood. In tissue culture, the endothelial cells form continuous tight junctions with their neighbors. This results in a cellular layer impermeable to protein tracers. When exposed to hyperosmolar solutions, in an attempt to mimic the conditions that open the blood-brain barrier in vivo, we found a reversible separation of the tight junctions between contiguous endothelial cells. No indication of activation of pinocytosis was observed. In vitro systems provide a novel approach for studying the function of the blood-brain barrier and allow for observations not possible with intact animals. PMID- 3028230 TI - The choroid plexus as a route from blood to brain. PMID- 3028232 TI - Glial-interstitial fluid exchange. PMID- 3028231 TI - Mechanisms of neuropeptide interaction with the blood-brain barrier. PMID- 3028233 TI - Potassium transport in astrocytes and neurons in primary cultures. PMID- 3028234 TI - Comparison of NIOSH and AIA methods for evaluating asbestos fibres: effects of asbestos type, mounting medium, graticule type and counting rules. PMID- 3028235 TI - Pathology consultation. Melanotic neuroectodermal tumor of infancy. AB - The melanotic neuroectodermal tumor of infancy is a usually benign neuroectodermally derived tumor with a predilection for the anterior maxilla of children who are less than 1 year of age. The lesion is considered to be readily treated by conservative excision, but it should be regarded as potentially malignant, even though there is infrequent expression of such malignancy. PMID- 3028237 TI - [Proliferative activity and storiform structure of histiocytofibromas]. PMID- 3028236 TI - [Angioma-like cutaneous metastasis disclosing a hepatocarcinoma]. PMID- 3028238 TI - [Epidemic erythema infectiosum or fifth disease]. PMID- 3028239 TI - Stereotactic Bragg peak proton beam radiosurgery for cerebral arteriovenous malformations. AB - A series of 709 patients with inaccessible AVMs treated with the proton beam is reported. There is a follow-up time of over 2 years for 92% of the patients. AVMs have slightly lower thresholds for proton injury than does normal brain tissue. Proton beam therapy involves very low risk. However, the risks of treatment failure grow with inadequate therapeutic doses. PMID- 3028240 TI - Bleeding of a venous hemangioma inside the filum terminale. AB - Spinal vascular malformations can present with acute symptoms suggesting spinal cord or nerve root compression. We describe a case of bleeding from a venous hemangioma within the filum terminale. This malformation caused acute compression of the lumbar spinal roots. Thus, vascular malformations may exist behind the symptoms of acute radiating pain and lower extremity weakness. They can be treated by total extirpation, when possible, by resection or by decompressive laminectomy alone. PMID- 3028241 TI - Preliminary experiences with CO2-laser and ultrasonic aspirator in brain surgery. AB - The basic problems and potential of CO2-laser and ultrasound aspirator in brain surgery are presented on the basis of experience with the Sharplan 733A CO2-laser and the Sonotec ultrasound aspirator. Complete removal of meningiomas and gliotic scar and partial removal of malignant astrocytomas by CO2-laser and ultrasound aspirator was performed. There were no complications related to the surgical procedure. The main advantage of the CO2-laser was precise cutting of neural tissue and rapid evaporation for debulkment of tumor. The ultrasound aspirator was especially useful for rapid internal debulkment by fragmentation and aspiration of soft tumors. On the basis of these experiences the surgical theater should be equipped with these and other instruments in such a way that they can be readily and interchangeably used in any neurosurgical procedure. PMID- 3028242 TI - [Polycythemia in liver diseases]. PMID- 3028243 TI - Japanese encephalitis: identification of inflammatory cells in cerebrospinal fluid. AB - Fifteen patients with acute encephalitis were studied during the epidemic season of Japanese encephalitis (JE) in Northern Thailand; 13 of 15 proved to have the encephalitis. In serial cerebrospinal fluids and blood samples, mononuclear cells were identified using monoclonal antibodies. In cerebrospinal fluid from the patients with Japanese encephalitis, T cells predominated with a 4.2:1 ratio of T helper/inducer cells to T suppressor/cytotoxic cells; B cells and macrophages were often present but in small numbers compared to their presence in blood. Distribution of cell types did not vary between the first and fifth day of hospitalization, were similar in fatal and nonfatal cases, and were unaffected by administration of steroids. PMID- 3028244 TI - Molecular genetics of transposable elements in plants. PMID- 3028245 TI - The myc oncogene: its role in transformation and differentiation. PMID- 3028246 TI - Mechanism of bacteriophage mu transposition. PMID- 3028247 TI - Adenovirus promoters and E1A transactivation. PMID- 3028248 TI - Control of multicellular development: Dictyostelium and Myxococcus. PMID- 3028249 TI - Intergeneric and interspecies gene exchange in gram-positive cocci. PMID- 3028250 TI - Effect of antiviral agents on replication of herpes simplex virus type 1 in brain cultures. AB - An in vitro tissue culture system consisting of reaggregated embryonic brain cells was used to evaluate the inhibition of herpes simplex type 1 (HSV-1) by several antiviral compounds. The efficacy of acyclovir, vidarabine, bromovinyldeoxyuridine, and 9-(1,3-dihydroxy-2-propoxymethyl) guanine in HSV-1 infected Vero cell monolayer cultures was compared with that seen with brain cell aggregates. At a mean 50% inhibitory dose with Vero cells, acyclovir showed a 99% reduction of virus titer in brain cell aggregates. Vidarabine and 9-(1,3 dihydroxy-2-propoxymethyl) guanine gave a dose-dependent reduction in virus titer with Vero cells; however, in aggregate cultures treated with the same drugs a dose-dependent decrease at 24 h was followed by an increase to a point of no inhibition at 72 h postinfection. Pretreatment of brain cell aggregates with a hybrid human leukocyte interferon (Le IF-AD) reduced virus titers at 48 h postinfection but did not maintain this reduction at 72 h. In contrast, infected Vero cell monolayer cultures demonstrated a dose-dependent reduction in virus titers with Le IF-AD. Postinfection treatment with Le IF-AD did not reduce plaque formation in Vero cells but was effective in reducing virus titer in HSV-1 infected brain cell aggregates at 48 h postinfection. Antiviral concentrations of up to 200 micrograms or 200,000 IU/ml for interferon did not appear morphologically toxic to brain cells. Antiviral therapy of HSV-1-infected brain cell aggregates may more closely mimic in vivo responses than monolayer cultures. PMID- 3028252 TI - Microplate phosphocellulose binding assay for aminoglycoside-modifying enzymes. AB - We modified the phosphocellulose binding assay for aminoglycoside-modifying enzymes (AMEs) by use of microdilution plates and a multichannel micropipette. Batteries of aminoglycoside substrates for screening organisms for the presence of AMEs as well as for subclassifying enzymes were prepared and stored in microdilution plates. When tested in parallel with the conventional tube reaction assay, the microplate assay yielded comparable radioactive counts and therefore equally correct identifications of AMEs in 32 isolates representing nine bacterial species. Other modifications, such as multichannel dispensing of crude enzyme preparations and radioisotopic precursors, provided a more rapid, convenient, and less expensive means of examining large collections of organisms for AMEs. PMID- 3028251 TI - Comparison of two beta-lactamase-producing strains of Streptococcus faecalis. AB - A second strain of enterococcus (PA) producing beta-lactamase (Bla+ phenotype) was compared with the previously reported Bla+ enterococcus, strain HH22. As with the original strain, there was a marked inoculum effect when PA was tested with penicillin, ampicillin, and piperacillin; no difference was noted with methicillin, cephalothin, imipenem, or vancomycin; the difference with ticarcillin was intermediate. High-level gentamicin resistance (Gmr) transferred from PA to an enterococcal recipient strain at a frequency approximately 100-fold lower than for HH22; all Gmr transconjugants from both strains were Bla+, but only PA showed linkage of Gmr and Bla+ with transfer of resistance to streptomycin, tetracycline, and chloramphenicol. EcoRI digestion of plasmid DNA from Gmr Bla+ transconjugants showed no similarities between the two strains. A 5.1-kilobase EcoRI Bla+-encoding fragment derived from HH22 was cloned into an Escherichia coli cloning vector and shown to hybridize to a 10.2-kilobase EcoRI fragment derived from PA; both fragments hybridized to an 840-base-pair staphylococcal Bla+ gene probe. These data indicate that the penicillinases are similar but encoded on different or differently arranged plasmids. The fact that both are transferable emphasizes the potential for this new streptococcal resistance determinant to disseminate. PMID- 3028254 TI - Antistaphylococcal activity of a cyclic peptide, LY146032, and vancomycin. AB - The inhibitory and bactericidal activities of a novel cyclic peptide antibiotic, LY146032, and vancomycin against oxacillin-susceptible and oxacillin-resistant staphylococci were compared. The MICs for 90% of strains were two- to fourfold higher for vancomycin than for LY146032. MBC/MIC ratios for all strains were less than or equal to 2. In killing rate studies with four strains of staphylococci, there was no detectable growth in the presence of 4X the MIC of either LY146032 or vancomycin after 24 h of incubation. PMID- 3028253 TI - Involvement of outer membrane of Pseudomonas cepacia in aminoglycoside and polymyxin resistance. AB - Pseudomonas cepacia was found to be resistant to the outer membrane permeabilizing effects of aminoglycoside antibiotics, polymyxin B, and EDTA. Permeabilization of P. cepacia to the fluorescent probe 1-N-phenylnaphthylamine was not achieved at concentrations 100- to 1,000-fold above those required to permeabilize Pseudomonas aeruginosa. Furthermore, in contrast to P. aeruginosa cells, intact cells of P. cepacia did not bind the fluorescent probe dansyl polymyxin. However, purified lipopolysaccharide (LPS) from P. cepacia bound dansyl-polymyxin with approximately the same affinity as did LPS from P. aeruginosa. Also, binding of dansyl-polymyxin to P. cepacia (and P. aeruginosa) LPS was inhibited by polymyxin B, streptomycin, gentamicin, and Mg2+. These data suggest that P. cepacia does not utilize the self-promoted pathway for aminoglycoside uptake and that the outer membrane is arranged in a way that conceals or protects cation-binding sites on LPS which are capable of binding polycations such as aminoglycosides or polymxyin. PMID- 3028255 TI - Mechanism of the cyanide-catalyzed oxidation of alpha-ketoaldehydes and alpha ketoalcohols. AB - Cyanide catalyzes the reduction of dioxygen or of ferricytochrome c by dihydroxyacetone phosphate. The rapid initial phase of these reactions, but not the subsequent slow phase, was augmented by incubating the triose phosphate aerobically or anaerobically at pH 9.0 prior to adding the cyanide. The aerobic incubation, which was most effective, was associated with a decline in enediol, whereas the less effective anaerobic incubation was accompanied by an increase in enediol content. This suggested that the alpha-ketoaldehyde product of autoxidation of the enediol, rather than the enediol itself, was responsible for the rapid phase reaction which followed addition of cyanide. This was confirmed by exploring the cyanide-catalyzed oxidation of the alpha-ketoaldehyde, phenylglyoxal. The inhibitory effect of the manganese-containing superoxide dismutase indicated that O2- was a kinetically important intermediate of the rapid phase reaction. A reaction mechanism is proposed which is consistent with the results presented. PMID- 3028256 TI - Subfractionation of the outer membrane of rat brain mitochondria: evidence for the existence of a domain containing the porin-hexokinase complex. AB - Isolated and well-characterized rat brain nonsynaptic mitochondria were subfractionated by digitonin. Antibodies to a uniquely outer membrane protein, porin, have allowed us to use this protein for the first time as an outer membrane marker in brain. Hexokinase, which binds to porin, was also measured. Based upon the sequential release of these and other marker enzymes with increasing concentrations of digitonin, three outer membrane domains have been identified. Two populations of porin were found by this treatment. The most plausible interpretation of our results is that the two porin populations exist in different membrane environments with regard to cholesterol. One of these populations binds most of the hexokinase and appears to be associated with the inner membrane. It is proposed that the porin-hexokinase complex in brain mitochondria is located in a cholesterol-free membrane domain together with inner membrane components. This domain has the features of contact points which have been visualized by electron microscopy. PMID- 3028258 TI - Methane monooxygenase: purification and properties of flavoprotein component. AB - An anaerobic procedure was developed for the purification of the flavin:NADH oxidoreductase (flavoprotein) component of methane monooxygenase to homogeneity. The molecular weight of the flavoprotein determined by gel filtration was about 40,000, and by sedimentation equilibrium analysis, about 38,000. The purified flavoprotein is a monomeric protein with a sedimentation constant (S20,W) value of about 2.1 S. The absorption spectrum of the flavoprotein has a peak at 460 nm and shoulder at 395 nm. The fluorescent excitation and emission spectra of the fluorescent component of flavoprotein had peaks at 450, 370, and 530 nm, respectively. A FAD was identified as a prosthetic group of flavoprotein by thin layer chromatography. The flavoprotein contained about 1 mol of FAD and 2 mol each of iron and acid-labile sulfide per mole of protein. The flavoprotein was directly reduced by NADH under anaerobic conditions. The formation of neutral flavin semiquinone was detected during anaerobic titration of flavoprotein by NADH and also as a free radical signal at a g value of 2.004 by EPR spectroscopy. The iron sulfur cluster has g values of 2.04, 1.96, and 1.87, yielding a g average of 1.96, characteristic of a Fe2S2 center. Antibody prepared against the flavoprotein reacted with flavoprotein and inhibited methane monooxygenase activity. PMID- 3028259 TI - On the mechanism of chlorination by chloroperoxidase. AB - Spectral-scan results obtained on the millisecond time scale are reported for reactions of chloroperoxidase with peracetic acid and chloride ion in both the presence and the absence of monochlorodimedone. A multimixing experiment is performed in which stoichiometric amounts of chloroperoxidase and peracetic acid are premixed for 0.7 s before the resultant compound I is reacted with chloride ion. The combined results show that the only detectable enzyme intermediate species is compound I (except in very late stages of the reaction), that the disappearance of compound I is accelerated by the presence of chloride ion, and that it is further accelerated if both chloride and monochlorodimedone are present. It is concluded that compound I is an obligate intermediate species in the reaction. Experiments are performed on the reaction of monochlorodimedone with hypochlorous acid in both the presence and the absence of added chloride ion, but in the absence of chloroperoxidase. The presence of chloride ion greatly accelerates the reaction rate apparently by setting off a chlorine chain reaction. This reaction would be important in the enzyme-catalyzed reaction if hypochlorous acid were liberated into the solution. A careful analysis of steady state kinetic results shows that in the chlorination of monochlorodimedone at least, liberation of free hypochlorous acid is not important in the enzyme catalyzed pathway. Rather the reaction proceeds from compound I to formation of iron(III)-OCl by chloride ion addition to the ferryl oxygen atom. This obligate intermediate species then chlorinates the substrate. It is well described as enzyme-activated hypochlorous acid, in which replacement of the proton in HOCl by the heme iron ion produces a Cl+ species of great potency. Thus the enzyme controls chlorination of monochlorodimedone rather than unleashing an uncontrolled chain reaction in which it would be rapidly destroyed. PMID- 3028260 TI - Formation of glutathione-conjugated semiquinones by the reaction of quinones with glutathione: an ESR study. AB - The nonenzymatic reaction of the cytotoxic compounds menadione (2-methyl-1,4 naphthoquinone) and 1,4-naphthoquinone (a reactive metabolite of 1-naphthol) with reducing agents such as NADPH and glutathione led to the formation of semiquinone free radicals, which were detected with electron spin resonance spectroscopy. In the presence of glutathione as a reducing agent, menadione and 1,4-naphthoquinone underwent net one-electron reduction and conjugation with glutathione. At higher concentrations of glutathione, 1,4-naphthoquinone formed the semiquinones of both the monoconjugate and the diconjugate. The naphthoquinone-glutathione conjugates should redox cycle in a manner already known for the menadione conjugate. The semiquinone intermediates could be detected only under a nitrogen atmosphere and are probably the primary oxygen-reactive species responsible for the redox cycling of menadione- and naphthoquinone-glutathione conjugates. PMID- 3028257 TI - The molecular dynamics of actin measured by a spin probe attached to lysine. AB - Rabbit skeletal muscle G-actin was labeled with a spin probe, 3-(5-fluoro-2,4 dinitroanilino)proxyl. Tryptic digestion of the labeled actin followed by ultrafiltration and ion-exchange column chromatography indicated that the label was attached to residue Lys-61. This residue is found within a 9-kDa N-terminal segment that is easily degraded by proteolytic enzymes. The rate of reduction of the nitroxide bond by ascorbate was measured to determine the accessibility of the probe to small molecules in the solvent. These experiments showed that label bound to G-actin was relatively inaccessible to ascorbate, suggesting that it is buried within the protein structure. Polymerization further decreased the accessibility of the probe. Replacing bound Ca2+ with Mn2+ decreased the observed intensity of the electron paramagnetic resonance signal, indicating the spin label is about 2 nm distant from the metal binding site on the actin molecule. Labels attached to G-actin displayed an absorption spectrum characteristic of rotational motion with a correlation time (tau c) of 7 X 10(-9) s, which is faster than that for the whole molecule. Labels attached to F-actin had a value of tau c, measured using saturation transfer electron paramagnetic resonance, of 2 X 10(-5) s, which shows that the probe has a greater degree of mobility than the filament. The binding of heavy meromyosin or troponin-tropomyosin to labeled actin resulted in a further increase in the rotational correlation times, with the greatest decrease in mobility (tau c = 1 X 10(-4) s) observed when both were bound. Together the above results suggest that the 9-kDa segment of actin is mobile relative to the rest of the molecule and that this mobility can be influenced by the binding of heavymeromyosin or troponin-tropomyosin. PMID- 3028262 TI - Role of protein kinase C in the regulation of rat liver glycogen synthase. AB - Rat liver glycogen synthase was phosphorylated by purified protein kinase C in a Ca2+- and phospholipid-dependent fashion to 1-1.4 mol PO4/subunit. Analysis of the 32P-labeled tryptic peptides derived from the phosphorylated synthase by isoelectric focusing and two-dimensional peptide mapping revealed the presence of a major radioactive peptide. The sites in liver synthase phosphorylated by protein kinase C appears to be different from those phosphorylated by other kinases. Prior phosphorylation of the synthase by protein kinase C has no significant effect on the subsequent phosphorylation by glycogen synthase (casein) kinase-1 or kinase Fa, but prevents the synthase from further phosphorylation by cAMP-dependent protein kinase, Ca2+/calmodulin-dependent protein kinase, phosphorylase kinase, or casein kinase-2. Additive phosphorylation of liver glycogen synthase can be observed by the combination of protein kinase C with the former set of kinases but not with the latter. Phosphorylation of liver synthase by protein kinase C alone did not cause an inactivation nor did the combination of this kinase with glycogen synthase (casein) kinase-1 or kinase Fa produce a synergistic effect on the inactivation of the synthase. Based on these findings we conclude that the phorbol ester induced inactivation of glycogen synthase previously observed in hepatocytes cannot be accounted for entirely by the activation of protein kinase C. PMID- 3028263 TI - Generation of allantoin from the oxidation of urate by cytochrome c and its possible role in Reye's syndrome. AB - Allantoin in the presence of calcium ions has been implicated as a potential toxic agent in Reye's syndrome. An investigation of possible alternative sources of allantoin in humans, which lack the enzyme uricase, has been initiated. Urate is a strong reducing agent which can reduce cytochrome c nonenzymatically, with the concomitant production of CO2 and H+. The stoichiometries measured for the various reactants and products were 1 urate:2 cytochrome c:1 H+:1 CO2. The initial reaction rate depended on the concentrations of both urate and cytochrome c, with reaction kinetics that were first order with respect to urate and second order with respect to cytochrome c. The participation of molecular oxygen in this reaction could not be detected. The pH and ionic strength optima for this reaction were determined to be 9.5-10.5 and 10(-5) M, respectively. Based on the results reported here, the following balanced equation can be written: urate-2 + 2 cytochrome c+3 + 2 H2O----allantoin + 2 cytochrome c+2 + H+ + HCO3-. We propose that allantoin can be generated from the oxidation of urate by cytochrome c+3, and that this is a potential source of allantoin in human tissues. PMID- 3028261 TI - The interaction of anions with native and phenylglyoxal-modified human serum transferrin. AB - The interaction of various anions with human serum transferrin was investigated due to the concomitant binding of iron and a synergistic anion to form the transferrin-anion-iron complex. Two tetrahedral oxyanion oxidizing agents, periodate and permanganate, were found to partially inactivate transferrin when used at equimolar ratios of oxidizing agent to protein active sites. Hypochlorite, a strong oxidizing agent with little structural similarity to periodate and permanganate, had little effect on iron-binding activity when used at similar low molar ratios of reagent to transferrin active sites. Transferrin treated with a 3:1 molar ratio of periodate or permanganate to active sites lost 74 or 67% of its iron-binding capacity, respectively. The composition of the buffer affected the extent of transferrin inactivation by periodate and permanganate; for example, the extent of inactivation by periodate was threefold greater in a borate buffer than in a phosphate buffer. Comparative oxidations in buffer systems suggest the following order of affinity of three buffer anions for the apotransferrin metal-binding center: phosphate greater than bicarbonate greater than borate. The interaction of phosphate ions with the iron-transferrin complex was also examined due to the increased susceptibility to periodate inactivation of iron-saturated transferrin in phosphate buffer (M. H. Penner, R. B. Yamasaki, D. T. Osuga, D. R. Babin, C. F. Meares, and R. E. Feeney (1983) Arch. Biochem. Biophys. 225, 740-747). The apparent destabilization of the iron transferrin complex in phosphate buffer was found to be due to the competitive removal of iron by phosphate from the iron-protein complex. We found that phenylglyoxal-modified Fe-transferrin, with no loss of bound iron, was much more resistant to iron removal by phosphate and other competitive chelators. PMID- 3028264 TI - Structure and function of the Ah receptor: sulfhydryl groups required for binding of 2,3,7,8-tetrachlorodibenzo-p-dioxin to cytosolic receptor from rodent livers. AB - Cytosol from rodent liver was exposed to a variety of sulfhydryl-modifying reagents to determine if the cytosolic Ah receptor contained reactive sulfhydryl groups that were essential for preservation of the receptor's ligand binding function. At a 2 mM concentration in rat liver cytosol, all sulfhydryl-modifying reagents tested (except iodoacetamide) both blocked binding of [3H]2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD) to unoccupied receptor and caused release of [3H]TCDD from receptor sites that had been labeled with [3H]TCDD before exposure to the sulfhydryl-modifying reagent. Exposure of cytosol to iodoacetamide before labeling with [3H]TCDD prevented subsequent specific binding of [3H]TCDD, but iodoacetamide was not effective at displacing previously bound [3H]TCDD from the Ah receptor. The mercurial reagents, mersalyl, mercuric chloride, and p hydroxymercuribenzoate, were more effective at releasing bound [3H]TCDD from previously labeled sites than were alkylating agents (iodoacetamide, N ethylmaleimide) or the disulfide compound 5,5'-dithiobis(2-nitrobenzoate). Presence of bound [3H]TCDD substantially protected the Ah receptor against loss of ligand binding function when the cytosol was exposed to sulfhydryl-modifying reagents. This may indicate that the critical sulfhydryl groups lie in or near the ligand binding site on the receptor. Subtle differences exist between the Ah receptor and the receptors for steroid hormones in response to a spectrum of sulfhydryl-modifying reagents, but the Ah receptor clearly contains a sulfhydryl group (or groups) essential for maintaining the receptor in a state in which it can bind ligands specifically and with high affinity. PMID- 3028265 TI - Calcium . calmodulin-dependent protein kinase II and calcium . phospholipid dependent protein kinase activities in rat tissues assayed with a synthetic peptide. AB - Rat tissue levels of Ca2+ . calmodulin-dependent protein kinase II (protein kinase II) and Ca2+ . phospholipid-dependent protein kinase (protein kinase C) were selectively assayed using the synthetic peptide syntide-2 as substrate. The sequence of syntide-2 (pro-leu-ala-arg-thr-leu-ser-val-ala-gly-leu-pro-gly-lys lys) is homologous to phosphorylation site 2 in glycogen synthase. The relative Vmax/Km ratios of the known Ca2+-dependent protein kinases for syntide-2 were determined to be as follows: protein kinase II, 100; protein kinase C, 22; phosphorylase kinase, 2; myosin light chain kinase, 0.005. Levels of protein kinase II were highest in cerebrum (3.36 units/g tissue) and spleen (0.85 units/g) and lowest in testis (0.05 units/g) and kidney (0.04 units/g). Protein kinase II activity was localized predominantly in the 100,000g particulate fraction of cerebrum and testis, in the supernatant fraction of heart, liver, adrenal, and kidney, and about equally distributed between particulate and supernatant in spleen and lung. Likewise, protein kinase C activity was highest in cerebrum (0.56 units/g) and spleen (0.47 units/g), and the majority of activity was present in the cytosolic fraction for all tissues measured except for cerebrum and testis in which the kinase activity was equal in both fractions. Finally, the ratios of protein kinase II to protein kinase C were different in various rat tissues and between particulate and supernatant fractions. These results suggest somewhat different functions for these two Ca2+-regulated, multifunctional protein kinases. PMID- 3028266 TI - Ferredoxin-thioredoxin reductase, an iron-sulfur enzyme linking light to enzyme regulation in oxygenic photosynthesis: purification and properties of the enzyme from C3, C4, and cyanobacterial species. AB - Ferredoxin-thioredoxin reductase (FTR), an enzyme involved in the light regulation of chloroplast enzymes, was purified to homogeneity from leaves of spinach (a C3 plant) and corn (a C4 plant) and from cells of a cyanobacterium (Nostoc muscorum). The enzyme is a yellowish brown iron-sulfur protein, containing four nonheme iron and labile sulfide groups, that catalyzes the activation of NADP-malate dehydrogenase and fructose 1,6-bisphosphatase in the presence of ferredoxin and of thioredoxin m and f, respectively. FTR is synonymous with the protein earlier called ferralterin. FTR showed an Mr of about 30,000 (determined by sedimentation equilibrium ultracentrifugation, amino acid composition, gel filtration, and gradient gel electrophoresis) and was composed of two dissimilar subunits (as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis). One of the FTR subunits from each source was similar both in Mr (about 13,000) and immunological properties, while the other subunit (of variable molecular weight) was characteristic of a particular organism. The similar subunit contained a disulfide group that was rapidly reduced by a dithiol (dithiothreitol) but not by monothiols (2-mercaptoethanol or reduced glutathione). Homogeneous FTR formed a tight noncovalent complex with ferredoxin on affinity columns. The basis for the structural variation in the different FTR enzymes remains to be determined. PMID- 3028267 TI - Phenobarbital-induced alterations in the activities of the transport and hydrolytic components of the glucose-6-phosphatase system in smooth and rough subfractions of the rat hepatic endoplasmic reticulum. AB - We have examined the influence of the phenobarbital-induced proliferation of the hepatic endoplasmic reticulum (ER) on the activities of the components of the glucose-6-phosphatase system, i.e., the enzyme, the glucose-6-P translocase (T1), and the phosphate translocase (T2). Young male rats were injected ip twice daily for 4 days with 4 mg/100 g body wt of phenobarbital (PB) or an equivalent volume of saline solution. On the fifth day, the rats were killed and smooth (SER) and rough (RER) fractions of the ER were isolated from liver homogenates. Kinetic constants for glucose-6-P hydrolysis by the system and enzyme were determined and used to calculate the kinetic constants for glucose-6-P transport. T2 activity was approximated by assaying the pyrophosphatase activity at pH 6.0 in intact microsomes. Three times more SER protein was recovered from livers of PB-treated rats. PB-treatment did not alter total liver enzyme activity, but total liver T1 activity was decreased to 59% of the control value. Maximal specific activities of the system, enzyme and T1 were all reduced by PB treatment to 44% of control values in the RER and to 68% of control values in the SER. PB treatment reduced the apparent activity of T2 in RER and SER to 35 and 49% of the respective control values. In the SER from both groups of rats, T1 activity or apparent T2 activity divided by enzyme activity was about 55% of the corresponding ratio in the RER. Our analysis of these data suggests that the lower activities of T1 and T2 in the smooth ER are the results of suppression by some intrinsic component localized in the smooth membrane. Accordingly, the reduction in total liver T1 activity and, therefore, system activity in PB-treated rats reflects the redistribution of the glucose-6-P translocase from the RER to the more abundant SER membrane where it is less active. The possibility is discussed that a higher cholesterol content within the SER membrane is responsible for the lower transport activities. PMID- 3028269 TI - Determination of dissolved oxygen in photosynthetic systems by nitroxide spin probe broadening. AB - Concentrations of dissolved oxygen were monitored by following the width of the midfield line of the electron spin resonance spectrum of a nitroxide spin-probe. Measurements of peak-to-trough widths of first derivative spectra yielded accurate data over a high range of O2 concentrations (up to 5mM). Continuous traces of second harmonic line heights yielded similar results and proved to be advantageous for kinetic measurements, but were nonlinear and less sensitive at O2 levels above 2 mM. Photosynthetic oxygen evolution and respiratory oxygen uptake in the cyanobacterium Agmenellum quadruplicatum were thus examined over a broad range of oxygen concentrations. Upon prolonged (greater than 1 min) illumination, effects of photooxidative damage to both photosynthesis and respiration were demonstrated in the same experimental system. With the addition of an impermeable paramagnetic broadening agent, rapid transients in intracellular concentrations of dissolved O2 also could be measured. PMID- 3028268 TI - Mitogen-stimulated release of inositol phosphates in human fibroblasts. AB - Mitogenic stimulation of quiescent human fibroblasts (HSWP) with a growth factor mixture (consisting of epidermal growth factor (EGF), insulin, bradykinin, and vasopressin) rapidly induces an increase in Na influx via a Ca-mediated activation of an amiloride-sensitive Na/H exchanger. Inositol phosphates (specifically inositol-1',4',5'-phosphate) have been implicated in mediating the mobilization of intracellular Ca stores in other cell types and we have now completed a detailed analysis of the mitogen-induced release of inositol phosphates in HSWP cells. Stimulation of inositol trisphosphate release is rapid (within 5 s) and reaches a maximum level (416-485% basal) within 10-15 s after the addition of growth factor mixture. Inositol bisphosphate and inositol monophosphate reach maximum levels by 30 s (1257% basal) and 60 s (291% basal), respectively. Levels of all three compounds then decay toward basal levels but remain elevated (150-350% of basal levels) after 10 min of incubation with mitogens. The effects of different combinations of these growth factors and of the bee venom peptide, melittin, have also been determined. We have also found that 8-(N,N-diethylamino)octyl-3,4,5-trimethoxybenzoate, which prevents the mitogen-induced rise in intracellular calcium activity and activation of Na influx, does not alter the mitogen-stimulated accumulation of inositol trisphosphate. In addition, the calcium ionophore A23187, which increases cytosolic Ca activity and induces a Na influx, does not stimulate the release of inositol trisphosphate. Assays performed in the presence of lithium, which inhibits inositol phosphate monophosphatase, promotes the prolonged and enhanced accumulation of inositol monophosphate. Treatment with the phospholipase inhibitor mepacrine or pretreatment with dexamethasone reduces the amount of inositol phosphates released upon mitogenic stimulation. Hence mitogenic stimulation of HSWP cells leads to the rapid stimulation of inositol phosphate release via a calcium-independent mechanism and suggests inositol trisphosphate as a candidate to mediate the release of intracellular calcium stores which is involved in the processes responsible for the activation of the Na/H exchanger. PMID- 3028270 TI - Sodium ion influx in proliferating lymphocytes: an early component of the mitogenic signal. AB - Stimulation of pig peripheral blood lymphocytes with concanavalin A (Con A) provoked a rapid increase (two- to threefold) in the rate of ouabain-inhibitable K+ uptake observable within 3-10 min of stimulation with mitogen. At least two phases can be distinguished in the activation of the Na+/K+ pump: the early phase (till 3 h) is characterized by an unaltered number of ouabain binding sites and the later phase (noted at 5 h) by an increased number of such sites. Both K+ efflux and influx increased to the same extent, thereby maintaining [K+]i at the same level as in resting cells (120 mM). Within 3 min of addition of mitogen, the rates of total and amiloride-inhibitable Na+ uptake went up two- and fourfold, respectively, thus resulting in rapid increase in [Na+]i from 20 to about 50 mM. Activation of the Na+/K+ pump was not observed when the cells were stimulated with Con A in low Na+ medium (9 mM), nor did the usual rise in [Na+]i occur. When monensin (30 microM), a Na+/H+ ionophore, was added to resting cells, an increase in both [Na+]i and active K+ uptake occurred in normal medium but not when cells were suspended in low Na+ isotonic buffer. Amiloride (500 microM), on the other hand, prevented both the Con A-induced increase in [Na+]i and the activation of the Na+/K+ pump. Despite complete inhibition of the Na+,K+-ATPase in the presence of ouabain (1 mM), Con A activated the amiloride-inhibitable Na+ uptake in the usual way. In mouse splenocytes stimulated with Con A, there was also a parallel rise in both [Na+]i and active K+ uptake but this took considerably longer to occur than was the case in pig peripheral blood lymphocytes. Increase in both ionic fluxes, the former passive and the latter active, is essential to the entry and maintenance of the cells in proliferative cycle. PMID- 3028271 TI - A tyrosine-specific protein kinase from Ehrlich ascites tumor cells. AB - A protein tyrosine kinase that phosphorylates both alpha and beta subunits of inactivated (Na+,K+)-ATPase from dog kidney was purified about 500-fold from Ehrlich ascites tumor cell membranes. The enzyme required divalent cations Mn2+, Mg2+, or Fe2+ but was inhibited by Cu2+ or Zn2+. The purified enzyme phosphorylated the beta subunit about five times faster than the alpha subunit of the (Na+,K+)-ATPase. The random polymer poly(Glu80Tyr20) was an excellent substrate while casein was only marginally phosphorylated. In contrast, the purified transforming gene product of Rous sarcoma virus phosphorylated all three substrates and the (Na+,K+)-ATPase was preferentially phosphorylated on the alpha subunit. The transforming gene product of Fujinami sarcoma visue and EGF receptor kinase from A431 cells phosphorylated (Na+,K+)-ATPase poorly whereas casein was an excellent substrate. The molecular weight of the partially purified protein tyrosine kinase from Ehrlich ascites tumor cells determined by gel filtration was about 60,000. One of two major phosphorylated phosphopeptides resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis had an Mr of 60 kDa, thus suggesting that it might be the autophosphorylated protein tyrosine kinase. A phosphatase that hydrolyzes phosphorylated histones or poly(Glu80Tyr20) was partially purified from the same membrane. PMID- 3028272 TI - Characterization of the in vitro stability of the rat hepatic receptor for 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). AB - The in vitro stability of the Ah receptor from rat hepatic cytosol was evaluated by [3H]TCDD binding studies, gel filtration, and sucrose density gradient ultracentrifugation. Thermal inactivation of unoccupied receptor followed first order kinetics between 5 and 40 degrees C, with an estimated Ea for inactivation of approximately 35 kcal/mol. Protease inhibitors did not reduce and dilution slightly increased the inactivation rate at 20 degrees C. Recovery and 20 degrees C stability decreased with increasing ionic strength. The TCDD-receptor complex was less susceptible to degradation at 20 degrees C, even in the presence of 0.4 M KCl. Specific binding was markedly pH dependent, with maximum recovery at 7.6. Analysis of the pH curve suggested that cysteine sulfhydryl groups may be involved in TCDD binding. Dithiothreitol (1 mM) maximized recovery and 20 degrees C stability, and addition of the thiol largely reactivated binding sites lost from cytosol prepared without it. Removal of low molecular weight components of cytosol by gel filtration resulted in a rapid 20 degrees C inactivation rate that could not be lessened by dithiothreitol. Glycerol (10% v/v) and EDTA (1.5 mM) maximized recovery of specific binding, but both decreased 20 degrees C stability in a concentration-dependent manner. Calcium chloride (4 mM) increased stability at 20 degrees C by approximately 20%, and retarded the characteristic shift in sedimentation coefficient from approximately 9 to approximately 6 S in high-salt sucrose gradients. The fact that sodium molybdate (20 mM) decreased recovery and 20 degrees C stability when dithiothreitol was present but slightly increased stability in its absence suggested an antagonism between the two compounds. Molybdate mitigated the inactivation induced by 0.4 M KCl, an effect which may be related to the observation of dual peaks in molybdate-containing high-salt sucrose gradients. These data indicate that thermal inactivation of the unoccupied rat hepatic Ah receptor primarily may be due to physical rather than enzymatic processes; (ii) sulfhydryl oxidation, removal of low molecular weight cytosolic components, and high ionic strength result in rapid rates of inactivation at 20 degrees C; and (iii) the large degree of protection conferred by TCDD binding implies a very tight ligand-receptor interaction, and as such accords with TCDDs extraordinary potency and persistence in producing its toxic and biochemical effects. PMID- 3028273 TI - Superoxide-dependent redox cycling of citrate-Fe3+: evidence for a superoxide dismutaselike activity. AB - Citrate-Fe3+, reportedly a physiological chelate, exhibits superoxide dismutaselike activity, as evidenced by the inhibition of xanthine oxidase dependent cytochrome c reduction; the dismutation of xanthine oxidase-generated superoxide to hydrogen peroxide and oxygen, and the enhanced disproportionation of potassium superoxide. The catalytic activity of citrate-Fe3+ corresponds, on a molar basis, to 0.03% of that of copper- and zinc-containing superoxide dismutase. Although weak, this activity enables citrate-Fe3+ to inhibit superoxide and ADP-Fe3+ -dependent peroxidation of extracted microsomal lipids. Also, the dismutase activity of citrate-Fe3+ interferes with its ability to promote lipid peroxidation. It is proposed that chelation of Fe3+ by citrate may represent a protective mechanism against the deleterious consequences of superoxide generation. PMID- 3028274 TI - Sulphatide content in a membrane fraction isolated from rabbit gastric mucosal: its possible role in the enzyme involved in H+ pumping. AB - The sulphatide content of vesicular membrane fraction from rabbit mucosal gastric microsomes was analyzed. This vesicular membrane fraction, in addition to a high sulphatide content, was enriched in an ouabain-insensitive (H+ + K+)-ATPase, a (Mg+2 + K+)-activated phosphatase, and a H+ pumping activity. The enzyme system involved in the process of acid secretion and the translocation of K+ was studied in these membrane preparations treated with arylsulphatase A, an enzyme that specifically hydrolyzes sulphatide. The results indicate that the breakdown of sulphatides of the vesicular membrane fraction inactivated both the (H+ + K+) ATPase activity and the H+ pumping. Both activities were partially restored by the sole addition of sulphatide. The K+-stimulated ouabain-insensitive phosphatase activity, suggested as a partial reaction of the (H+ + K+)-ATPase sequence, was unaffected by arylsulphatase. These results suggest that sulphatides may play a function in the high activity binding site for K+ of the enzyme involved in H+ pumping. PMID- 3028275 TI - [Targeting chemotherapy of hepatocellular carcinoma by arterial administration of anticancer agents dissolved in lipiodol]. AB - The lipid lymphographic agent, Lipiodol ultrafluid has been found to remain selectively in hepatocellular carcinoma. Using this characteristic nature of Lipiodol, a new targeting anticancer chemotherapy was devised. In order to achieve targeting anticancer chemotherapy and useful anticancer effects, anticancer drugs must be dissolved or suspended in Lipiodol and diffuse out from the Lipiodol gradually. Oily anticancer agents such as SMANCS dissolved in Lipiodol (SMANCS/Lipiodol), Mitomycin C in Lipiodol (MMC/Lipiodol), Aclarubicin in Lipiodol (ACR/Lipiodol) and a mixture of these were administered by catheterizing the celiac or hepatic artery under X-ray monitoring in 216 patients with hepatocellular carcinoma. Remarkable anticancer effects of this targeting chemotherapy were achieved, the serum AFP level and tumor size both showing a decrease in 91% of cases. The survival period of patients with unresectable hepatoma treated with the present protocol was definitely longer than the comparison group. PMID- 3028276 TI - [Oily chemoembolization of hepatoma]. AB - Since 1983 we have performed transcatheter oily chemoembolization (TOCE) using adriamycin (40-100 mg), Lipiodol (5-20 ml) and Gelfoam in the treatment of 100 cases with unresectable hepatocellular carcinoma. Adriamycin was dissolved in a fluid equal in specific gravity to Lipiodol and the adriamycin solution was mixed with 3 volumes of Lipiodol, making an adriamycin-in-oil emulsion (AOE). After TOCE, the blood concentration of adriamycin was obviously lower than that after one-shot injection because of the slow release of adriamycin from the AOE. Also, in cases of hepatic resection after TOCE, there was a clear difference in the adriamycin concentration between the tumor and the normal hepatic tissue. The cumulative survival rates for the 100 patients treated by TOCE were: 6 months 81.9%, 1 year 53.8% and 2 years 36.5%. Thus, improvement was found in comparison with the cumulative survival rates for 104 patients who underwent hepatic embolization without Lipiodol, which were 6 months 67.4%, 1 year 45.2% and 2 years 16.3%. AOE retained in the tumor as microemboli brings about the slow releasing effect of adriamycin. Furthermore, by adding the effect of Gelfoam embolization, TOCE has a strong antitumor effect. PMID- 3028277 TI - [Chemoembolization with degradable starch microspheres in malignant hepatic tumors]. AB - A new method of chemoembolization with degradable starch microspheres (DSM) was used for patients with malignant hepatic tumors. DSM, 40-45 micron in diameter, which are degraded by serum amylase, temporarily obstruct arterial blood flow at the arteriolar capillary bed. Experimental studies have demonstrated that such occlusion enhances the regional uptake and reduces systemic exposure to simultaneously administered arterial anticancer drugs. Transcatheter chemoembolization with DSM was performed in 14 cases of hepatocellular carcinoma and 8 cases of metastatic liver cancer. Adriamycin or Mitomycin C mixed with DSM was injected into the patients with hepatocellular carcinoma or metastatic liver cancer, respectively, through the proper hepatic artery. The therapeutic effect of this chemoembolization was evaluated by the change in tumor size measured by angiography or computed tomography. In hepatocellular carcinoma, tumor regression of over 50% was observed in 5 of 14 patients. Elevated serum AFP level of more than 200 ng/ml was decreased in all 6 cases. In metastatic liver cancer, tumor regression of over 50% was observed in 4 of 8 cases. Although half of the patients had transient pain within 2 hours, no major side effects such as bone marrow suppression and hepatotoxicity were observed. Our results suggest that chemoembolization with DSM can be effectively used in the treatment of malignant hepatic tumors. PMID- 3028278 TI - [Clinical evaluation of MMC-mc-chemoembolization therapy and its combination with UFT in hepatocellular carcinoma]. AB - A retrospective comparative study was done on the management of hepatoma: MMC-mc chemoembolization therapy (group A; 9 patients) vs. the same therapy in combination with UFT as a maintenance therapy (group B; 8 patients). The six month survival rate was 44.4% for group A, and 87.5% for group B. Generalized Wilcoxon test for the whole period revealed that the survival was favorable for group B as compared with A (P = 0.048). For adverse reactions in UFT therapy, no subjective signs occurred, but objective ones appeared such as one case of leukocytopenia, one of thrombocytopenia and 3 of increased GOT. However, no cessation of UFT therapy was necessary in these patients. These findings suggested that maintenance therapy with UFT may be favorable in this type of treatment for hepatoma. PMID- 3028279 TI - [A case of hepatocellular carcinoma with bone metastasis which responded to oral administration of UFT]. AB - A 61-year-old male was referred to our hospital for evaluation of right upper arm pain. He was diagnosed as having primary hepatocellular carcinoma with bone metastasis by his high titer (111,683 ng/ml) of serum alpha-fetoprotein, computed tomography and abdominal angiography, and so UFT therapy, 400 mg daily, was instituted. After 2 months of this therapy, the titer of serum alpha-fetoprotein gradually decreased. Seven months later, the titer was 3,997 ng/ml and reduction of the hepatic tumor size was shown by computed tomography and ultrasonography. Furthermore, the right upper arm pain diminished and X-ray examination revealed remarkable improvement. During chemotherapy, there was no severe side effect apart from mild anemia and the patient is presently still alive. This case suggests the efficacy of UFT for the treatment of primary hepatocellular carcinoma. PMID- 3028280 TI - What are adequate treatment and follow-up care for nonmelanoma cutaneous cancer? PMID- 3028281 TI - Human papillomavirus heterogeneity in 36 renal transplant recipients. AB - Immunosuppressed patients such as renal transplant recipients are prone to increased incidence of wart disease. We examined 48 tissue specimens from 36 renal transplant recipients using human papillomaviruses (HPVs) 1, 2, 3, 4, 5, and 6 in filter hybridization under stringent conditions. The results showed that 90% of the samples contained HPV DNA. Of these 43 positive samples, we found HPV 1 in 2%, HPV-2 in 56%, HPV-3 in 19%, HPV-4 in 47%, HPV-5 in 9%, and HPV-6 in 5%. In several cases, more than one type of HPV DNA was observed. In a few of these cases, the clinical appearance of the lesions differed from what might have been expected, such as those lesions containing HPV-3- or HPV-5-related DNAs. PMID- 3028282 TI - Human papillomavirus type 16 in bowenoid papulosis, intraoral papillomas, and squamous cell carcinoma of the tongue. AB - A 33-year-old immunosuppressed man developed bowenoid papulosis on his genitalia, velvety papules and plaques in his mouth, and invasive squamous cell carcinoma of his tongue. All three lesions were positive for human papillomavirus type 16 (HPV 16). The case provides further circumstantial evidence for a role of HPV-16 in epithelial neoplasia. The possible roles of a second HPV infection and of immunosuppression are also discussed. PMID- 3028283 TI - Spontaneous regression of perianal extramammary Paget's disease after partial surgical excision. AB - A 62-year-old woman presented with a three-year history of a pruritic perianal lesion, which was histologically confirmed to be perianal extramammary Paget's disease. Partial surgical excision of the lesion was followed by complete spontaneous regression of the residual plaque. PMID- 3028284 TI - Investigation of urinary tract infection. PMID- 3028286 TI - Experimental haemarthrosis produces mild inflammation associated with intracellular Maltese crosses. AB - Injection of autologous blood into rabbit joints induces an inflammatory reaction with Maltese cross-like birefringent spherulites. Similar microspherules seen in human joint fluids may be formed by lipids derived from breakdown of erythrocytes and other cells. PMID- 3028285 TI - Human papillomavirus DNA detected in squamous-cell carcinoma tissue of a patient with epidermodysplasia verruciformis. PMID- 3028287 TI - Deficient control of in vitro Epstein-Barr virus infection in patients with ankylosing spondylitis. AB - Infectious Epstein-Barr (EB) virus obtained from the B95-8 marmoset cell line was used to infect mononuclear cells from healthy controls and patients with rheumatoid arthritis (RA) and ankylosing spondylitis (AS), and outgrowth of B cells into lymphoblastoid cell lines was assessed by visual microscopy and uptake of tritiated thymidine over a 28 day period. When undiluted virus was used lymphocytes from both patients with AS and RA and from normal controls outgrew into lymphoblastoid cell lines (LCLs) by day 28 of culture. At dilutions of 1/10, 1/20, and 1/40, however, the control cells showed regression of proliferation at approximately day 14 of culture, whereas the cells from patients with AS and RA continued to proliferate and outgrew into LCLs (transformation scores of cells from patients with AS compared with controls at day 28 p less than 0.05 in all cases; thymidine uptake at a 1/40 dilution at day 28, patients with AS compared with controls p less than 0.01). Hence these results suggest that there is a defect in the cellular response to EB virus induced B cell proliferation in patients with AS similar to that seen with cells from RA donors. PMID- 3028288 TI - Collagenase production by human mononuclear cells in culture: inhibition by gold containing compounds and other antirheumatic agents. AB - Human peripheral blood mononuclear cells in adherent cultures have been shown to synthesise and secrete collagenase. In the present study we have examined the modulation of collagenase production in these cultures by several antirheumatic agents. Incubation of monocytes in serum free medium with sodium aurothiomalate in concentrations varying from 7.7 X 10(-7) to 7.7 X 10(-3) mol/l resulted in marked dose dependent inhibition of the collagenase production. This inhibition was apparently selective in that total protein synthesis or the viability of the cells were not affected. Similar inhibition of the collagenase production was also noted with auranofin, aurothioglucose, and chloroauric acid. The inhibition with auranofin was achieved with a concentration as low as 7.4 X 10(-8) mol/l. To examine the mechanisms of the inhibition of the collagenase activity induced by sodium aurothiomalate the production of prostaglandin E2 was also measured in the same cell cultures. Sodium aurothiomalate in concentrations greater than 7.7 X 10(-4) mol/l significantly inhibited the prostaglandin E2 production; the prostaglandin E2 production was not inhibited, however, in 7.7 X 10(-5) mol/l concentration, while the collagenase production was reduced by 51.0%. Also, exogenous prostaglandin E2 added to the cultures only slightly reversed the inhibition of the collagenase production by sodium aurothiomalate. Thus the inhibition of collagenase production by sodium aurothiomalate in human adherent mononuclear cell cultures appears to be independent of the inhibition of prostaglandin E2 production. The inhibition of collagenase produced by monocyte macrophages, as shown here in vitro, may contribute to the clinical efficacy of the compounds tested in the treatment of rheumatoid arthritis. PMID- 3028289 TI - Depressed superoxide radical generation by neutrophils from patients with rheumatoid arthritis and neutropenia: correlation with neutrophil reactive IgG. AB - Neutrophils of 31 patients with neutropenia and rheumatoid arthritis (RA) have been studied to assess their ability to generate superoxide radicals (O-2) on activation. Seventeen patients had classical Felty's syndrome and 14 presumed chrysotherapy related neutropenia. Results were compared with those from age and sex matched controls with uncomplicated RA and from normal subjects. Neutrophils from patients with Felty's syndrome had a significantly reduced ability to generate superoxide radicals when compared with the other three groups. In addition, serum levels of IgG polymorphonuclear leucocyte binding activity (IgG PBA) were also raised in the group with Felty's syndrome. A statistically significant inverse correlation existed between O-2 generation and IgG PBA. It is concluded that neutrophil reactive IgG may have an important role in both quantitative and qualitative defects in neutrophil function in Felty's syndrome. PMID- 3028290 TI - Malignant fibrous histiocytoma of the pulmonary artery. AB - A patient is reported whose symptoms and clinical features were interpreted as pulmonary embolism. However, the ultimate diagnosis was malignant fibrous histiocytoma of the pulmonary artery, which was made evident at operation. Although extremely rare in the early medical literature, primary sarcoma of the pulmonary artery has been reported more frequently during the last decade. This patient may provide further insight into the clinical, diagnostic, and therapeutic features of primary sarcomas of the pulmonary artery. PMID- 3028291 TI - Hyperlipoproteinemia, atherosclerosis risk, and dietary management. AB - There is new impetus for dietary intervention in response to the recent data relating levels of circulating lipids and lipoproteins to atherosclerosis risk. Certain levels of cholesterol, stratified for age and sex, are associated with "moderate" and "high" risk of atherosclerosis. Knowing the relationship of diet to lipid levels and lipid transport by the various lipoproteins enables the appropriate diagnosis and management. Dietary measures include attention to the level of calories and fat, the type of fat, cholesterol content, source of protein, the type of carbohydrate and fiber, and the level of alcohol intake. Lipid (cholesterol) screening at intervals is recommended for all adults and for children at special risk of hyperlipidemia. PMID- 3028292 TI - Activity of Na+ and K+ ATPase in the red blood cells of patients with septic shock. PMID- 3028293 TI - The paradoxical response to TRH in Cushing's disease is not a reliable diagnostic test. PMID- 3028294 TI - Temporal course of metabolic acidosis during cardiorespiratory arrest in cats. Treatment with sodium bicarbonate. PMID- 3028296 TI - [Nourseothricin--properties, biosynthesis, production]. PMID- 3028295 TI - Binding and activity of Bacillus thuringiensis delta-endotoxin to invertebrate cells. AB - Fluorescein isothiocyanate was used as a label to detect delta-endotoxin of Bacillus thuringiensis subsp. thuringiensis and israelensis in binding studies with different in vitro cell systems. Protoxin of the subspecies thuringiensis could be labelled directly whereas the activated toxin had to be traced indirectly with labelled antibodies. Both protoxin and activated toxin bound to primary midgut cell cultures of Pieris brassicae larvae as well as to cells of an established culture of Drosophila melanogaster. No binding with either toxin form could be observed with hemocytes of P. brassicae. Biological activity as shown by the trypan blue viability assay was obtained only with the activated toxin against the midgut cells. Toxin of the subspecies israelensis reacted very unspecifically. Binding followed by rapid destruction was obtained with all the tested cultures. PMID- 3028297 TI - [Development of the resistance of Escherichia coli to streptothricins (nourseothricin, grisin)]. PMID- 3028298 TI - [Ecologic studies of the nourseothricin resistance of coliform bacteria of intestinal flora in humans and animals]. PMID- 3028299 TI - [Mechanism of action of nourseothricin and bacterial resistance and cross resistance to this antibiotic]. PMID- 3028300 TI - [Antiviral effect of nourseothricin]. PMID- 3028301 TI - [Biologic determination of nourseothricin]. PMID- 3028302 TI - [Chemical determination of the streptothricin antibiotic nourseothricin]. PMID- 3028303 TI - [Pharmacokinetics of nourseothricin in laboratory animals]. PMID- 3028304 TI - [Pharmacologic action profile of nourseothricin]. PMID- 3028305 TI - [Immunoregulatory effect of nourseothricin sulfate]. PMID- 3028306 TI - [Acute and subchronic toxicity of nourseothricin in laboratory animals]. PMID- 3028309 TI - [Effect of the antibiotic nourseothricin on feeding, weight gain and feed utilization in piglets and fattening pigs and energy and protein deposition in fattening pigs]. PMID- 3028308 TI - [Long-term toxicity of nourseothricin]. PMID- 3028307 TI - [Local tolerance of nourseothricin]. PMID- 3028310 TI - [Use of nourseothricin in the raising of calves]. PMID- 3028311 TI - [Determination of the ergotropic effectiveness of the antibiotic nourseothricin in the raising of broiler chickens and its effect on the crude nutrient content of the carcass and length of the villi of the small intestine]. PMID- 3028312 TI - [Rhinovirus and acute respiratory infections in infants]. AB - Human leucocytary interferon (IFN) in a concentration of 2,000 Ul/ml stops the development of 9 Rhinovirus (RV) strains. The use of serum IFN antibodies (SAIF) reverses this effect. Of 256 nasal aspirates culture negative for other respiratory viruses, 40 RV were isolated with SAIF as opposed to 19 without serum anti IFN. The frequency and some clinical aspects are emphasized in infants: 43% had bronchiolitis similar to respiratory syncytial virus (RSV) infections and atopy was frequent. PMID- 3028313 TI - Purkinje cell inclusions and 'atelencephaly' in 13q-chromosomal syndrome. AB - Severe microcephaly was present from birth in a child with a 13q-chromosomal syndrome [46,XY,del(13)(q22q31)]. He died at 20 months of age. Neuropathologic findings included atelencephaly and eosinophilic cytoplasmic inclusions in cerebellar Purkinje cells. Ultrastructurally, the inclusions consisted of stacks of parallel cisternae separated by electron-dense granular material. The relationship between these inclusions and the smaller cytoplasmic inclusions known as "lamellar bodies" is discussed, and the central nervous system malformations in this syndrome are reviewed. PMID- 3028314 TI - Liver cell dysplasia and hepatocellular carcinoma in non-A, non-B hepatitis. AB - Liver cell dysplasia (LCD) is a premalignant cytologic change of hepatocytes that has been statistically linked to cirrhosis, hepatocellular carcinoma (HCC), and chronic liver disease related to hepatitis B virus. The relationship of LCD to non-A, non-B (NANB) hepatitis is currently unknown. We studied liver biopsy and surgical resection specimens from 36 patients with NANB hepatitis, and identified LCD in 17 (42.5%) of 40 specimens, most often associated with cirrhosis. Dysplasia was present in individual hepatocytes, in clusters, and in a distinctive "spreading" pattern of hepatocytes about central veins. Three patients had HCC with a predominant giant cell pattern, as well as LCD. These findings suggest that LCD and HCC should be included among the potential pathologic sequelae of NANB hepatitis. PMID- 3028315 TI - Hepatocellular carcinoma developed on noncirrhotic livers. Sinusoids in hepatocellular carcinoma. AB - In hepatocellular carcinoma, there is modification of cell-to-cell and cell-to extracellular matrix interactions. Two cases of well-differentiated hepatocellular carcinoma that developed in noncirrhotic livers were explored by light and electron microscopy on perfusion-fixed liver biopsy specimens. In addition, immunocytolocalization of collagen types I, III, and IV, laminin, and fibronectin was assessed. The main features included the following: absence of collagen staining inside the tumor with Sirius red; decrease of collagen types I and III and the increase of collagen type IV, laminin, and fibronectin; widening of Disse's spaces containing numerous, discontinuous, and thick fragments of basement membrane-like material piled up beneath endothelial cells and around perisinusoidal cells; transformation of perisinusoidal cells into cells with the characteristics of fibroblasts and myofibroblasts; decreased numbers of fenestrae for endothelial cells with processes often overlapping and attached with tight junctions; and rarefaction of Kupffer's cells. Expression of cellular and extracellular material abnormalities are related to the tumor differentiation. The study of these abnormalities may have implications in prognosis. PMID- 3028316 TI - Surgery in patients with acquired immunodeficiency syndrome. AB - Between 1982 and 1985, 21 patients with acquired immunodeficiency syndrome (20 men and one woman; mean age, 36 years) underwent 31 surgical procedures at the Harbor/UCLA Medical Center, Torrance, or the UCLA Medical Center (skin, lymph node, and endoscopic biopsies were excluded). The operations included seven emergencies and 24 elective operations (eight major and 16 minor). Pathologic findings included cytomegalovirus colon perforation (two), disseminated Kaposi's sarcoma (KS) of the small and large bowel (one), cystic duct obstruction by KS (one), poorly differentiated gastrointestinal lymphoma (one), Candida acalculous cholecystitis (one), central nervous system toxoplasmosis (two), amebic encephalitis with abscess (one), staphylococcal botryomycosis of the pericardium (one), pulmonary KS (one), and cytomegalovirus (one). The overall operative (30 days) mortality rate was 48% (10/21). The emergency surgery rate was 57% (4/7), elective, 43% (6/14). The high operative mortality rate in these patients was usually due to progression of opportunistic infections or malignancy. PMID- 3028317 TI - Neutrophil function in a rat model of endotoxin-induced lung injury. AB - Polymorphonuclear neutrophil leukocytes (PMNs) are known to cross the alveolar capillary barrier and enter the alveolus in acute adult respiratory distress syndrome (ARDS). The pathogenic role of PMNs in both the acute lung injury and subsequent infectious susceptibility in ARDS is not clear. In the present study we investigated the functional status of various neutrophil populations using a chronic, endotoxemia-induced ARDS model. Rats infused with Escherichia coli endotoxin for three days develop an acute lung injury with a histologic picture closely resembling human ARDS. The PMNs recovered from the circulation and by bronchoalveolar lavage were compared with normal rat PMNs. In endotoxemic animals, superoxide production was markedly enhanced in circulating PMNs, indicating production of high levels of potentially cytotoxic oxygen intermediates, while myeloperoxidase activity was decreased in both circulating and lavage PMNs, indicating depressed myeloperoxidase-dependent antimicrobial activity. PMID- 3028318 TI - [Nitrogen exchange in the large intestine of ruminants. 4. Exchange and isotope balance of intracecally administered 14C- and 15N-urea in sheep with simultaneous intracecal dose of a partly hydrolyzed straw meal]. AB - Two experiments were performed with wethers (Body weight 34 to 44 kg) receiving a ration rich in crude fibre at maintenance level. The animals were fitted with ileocaecal cannulas into which 14C-, 15N-labelled urea together with digesta was introduced hourly for a 24 hours period (V1; 2 animals). In experiment two (V2; 3 animals) in addition HCl-partly hydrolysed straw meal was introduced. After ureolytic degradation the intracaecal applied urea entered mainly the intermediary metabolism. The resulting ammonia was resynthesized to urea without any time lag. The rate constant for the increase in 15N labelling of urea was 3.2 d-1 in both experiments. Urea leaves the plasma with half lives of 10.6 (V1) and 5.2 (V2) hours. More than 60% of the applied urea were excreted with urine. Formed 14CO2 appeared at proportions of 66% (V2) and 71% (V1) in the respiration gases. Both, the decline of the 14C-activity in blood plasma and the specific 14C activity of CO2 in the respiration gases after the end of the labelling period do not follow a kinetic of first order. The 15N-labelling of the NH3-N in ileal digesta was very high and reached plateau values similar with those of plasma urea (2.54 vs. 2.56 atom-% 15N-excess). A direct entry of plasma urea into the small intestine was concluded. PMID- 3028319 TI - [Eating and ruminating behavior of sheep. 2. Experiments in pregnant and lactating ewes fed with a straw-concentrate mixture]. AB - Fertility oriented ewes in the late stage of gestation and during the lactation period received 19 feeding regimes on the basis of straw-concentrate mixtures, which essentially differed in the feedstuff structure. The ewes fed ad libitum on average used between 5 and 6 hours per day feed intake in the 86 measuring periods. The average daily ruminating time ranged from 398 to 502 minutes. An increasing straw content resulted in significantly longer ruminating periods and a higher number of masticatory movements, additional quotas of long straw had non effect. But even the most unfavourable feeding variants (straw-concentrate mixtures with 40% straw meal treated with NaOH) elicited 21 100 ruminating masticatory movements in 5 hours of ruminating. There were no significant differences between the absolute consumption and ruminating activities in the late stage of gestation and the lactation period; with reference to feed and crude fibre intake the ruminating expenditure in the late stage of gestation was always significantly higher. Rations with a low structural effect resulted in a more unbalanced distribution of consumption and ruminating activities in the course of the day than well structured rations. PMID- 3028320 TI - Control of some aspects of cis-platinum nephrotoxicity. AB - The elevations of blood urea nitrogen and serum creatinine caused by cis-platinum in rats can be prevented by a combination of procedures centered around the administration of sodium N-methyl-N-dithiocarboxyglucamine (NaG), before and very soon after the cis-platinum is administered in hypertonic (4.5%) saline. Elevations in-blood urea nitrogen and serum creatinine levels subsequent to such treatment are largely eliminated. These same procedures appear to have no effect on the anti-tumor action of the cis-platinum, as measured by tumor size and mass and by survival times, in female Sprague-Dawley rats inoculated with the Walker 256 carcinoma. The degree of myelosuppression, as measured by the white blood cell count is also slightly reduced. White blood cell counts returned to normal values more rapidly in animals treated with NaG than in those treated with cis platinum alone. An examination of the dose-response curve for the suppression of cis-platinum nephrotoxicity by NaG shows that this can be achieved with mole ratios of NaG: cis-platinum as low as 1:1 given after appropriate pretreatment. A preliminary structure-activity study on the suppression of cis-platinum induced elevations in blood urea nitrogen and serum creatinine by four closely related dithiocarbamates shows that this can also be achieved effectively by several dithiocarbamates in which the nitrogen atom bears polar substituents. PMID- 3028321 TI - Effects of individual terphenyls and polychlorinated terphenyls on rat hepatic microsomal cytochrome P-450-dependent monooxygenases: structure-activity relationships. AB - The effects of o-, m- and p-terphenyl, 2,4-dichloro-, 2,4,6-trichloro-, 2,3,5,6 tetrachloro-, 2,3,4,6-tetrachloro-, 2,4,4''',6- tetrachloro- and 2,3,4,5 tetrachloro-p-terphenyl, 2,3,4,5-tetrachloro-m- and o-terphenyl as inducers of hepatic drug-metabolizing enzymes were determined in immature male Wistar rats. o Terphenyl, 2,4-dichloro-, 2,4,6-trichloro-p-terphenyl and 2,3,4,5-tetrachloro-o terphenyl induced 4,4'-dimethylamino antipyrine N-demethylase at total dose levels of 300 mumol/kg and the 2,3,4,5-tetrachloro-p-terphenyl induced ethoxyresorufin O-deethylase (EROD). In contrast, none of the other terphenyls or polychlorinated terphenyls (PCTs) induced these enzyme activities. Previous studies have demonstrated that 2,3,4,5-tetrachloro-p-terphenyl did not exhibit a high affinity for the 2,3,7,8-tetrachlorodibenzo-p-trachlorodibenzo-p-dioxin (TCDD) receptor protein (EC50 = 6.6 X 10(-6) M). In contrast, this study showed that 2,3,4,5-tetrachloro-p-terphenyl was more active than either 2,3,4,5 tetrachloro-o- or m-terphenyl as an inducer of EROD. Moreover, the competitive receptor binding EC50 values for the latter two isomers were greater than 10(-5) M and this result was also consistent with their lack of EROD induction activity. Previous studies showed that analysis of the data for a series of 4'-substituted 2,3,4,5-tetrachlorobiphenyls indicated that the p-terphenyl structural moiety (i.e. 4'-substituent = phenyl) did not interact with high affinity with the receptor protein binding site. Since the 2,3,4,5-tetrachloro o- and m-terphenyls are also poor ligands for the receptor protein, this data and results from other studies indicate that PCT congeners (and commercial mixtures) are therefore unlikely to elicit significant 2,3,7,8-TCDD-like biologic or toxic effects in target species. PMID- 3028322 TI - Monitoring urine for inhaled cannabinoids. AB - The current position on the "passive smoking" of cannabis is reviewed with particular reference to the analysis of urine. The pharmacokinetics and metabolism of delta-9-tetrahydrocannabinol are first described, followed by a survey of methods used to identify and quantify its metabolites in urine. Published data concerning the appearance of cannabinoids in urine following "passive smoking" of cannabis are compared and the most important factors described. The problems of interpreting the results of the analysis of urine in forensic cases are discussed and a possible means to clarify the position by means of analysing serum is suggested. PMID- 3028323 TI - Interethnic differences in the detoxification of organophosphates: the human serum paraoxonase polymorphism. AB - Paraoxon, 0,0-diethyl-0-p-nitrophenylphosphate is the highly toxic metabolite of parathion. The activity of paraoxonase, the enzyme which hydrolyses paraoxon in human serum shows a genetically influenced polymorphism with strong interethnic differences. The serum paraoxonase genotype has a significant influence on the paraoxon clearance and consequently on the toxic action of paraoxon and some related organophosphates and definitively protects the serum cholinesterase. Persons with low paraoxonase activity seem to be more endangered when handling parathion and related insecticides. More than 50% of all Europeans can be included in this group. The distribution of paraoxonase activity in human serum will be shown for samples which were collected from all over the world. As one moves from Europe in the direction of Africa and Asia the percentage of the low activity group decreases and was not even demonstrable in some tribes. PMID- 3028325 TI - Cardiotoxicity of digitalis. AB - The principal effect of cardioactive glycosides (CG) is the inhibition of the (Na+ + K+)-ATPase system with subsequent increase in contractility of the myocardium. In subtoxic and toxic concentrations, CG increase O2 consumption due to a transient Ca2+ overload. Furthermore, the activity of several enzymes of the citrate cycle is changed; cAMP transiently rises with reduction of myocardial ATP, and intracellular lactate dehydrogenase and creatine kinase are lost in the coronary fluid. The antagonistic action of beta-receptor blocking agents is caused by their membrane-stabilizing effect. O2-consumption is increased in the non-failing heart, while in the failing one it decreased. The CG-induced arrhythmias are caused (1) by inhibition of the ATPase system of excitable cardiac structures, and (2) by interaction of CG with the autonomic nervous system. Severe intoxications and the rapid disappearance of cardiac symptoms upon administration of Fab fragments suggest that the CG-induced changes on the molecular level (with the exception of those on the ATPase system) are of secondary significance. PMID- 3028324 TI - DNA-mediated restoration of aryl hydrocarbon hydroxylase induction in a mouse hepatoma mutant defective in nuclear translocation of the Ah receptor. AB - In order to study the induction mechanism of aryl hydrocarbon hydroxylase (AHH), non-inducible mutants have previously been isolated from the mouse hepatoma cell line, Hepa-1. With the ultimate goal of isolating the corresponding genes, restoration of AHH inducibility to representative mutants by means of DNA mediated gene transfer has been set out. The successful transfection of a C- mutant, which is defective in nuclear translocation of the Ah receptor-inducer complex, is described here, using rat genomic DNA as donor material. Primary and secondary rat transfectants were obtained, and they were assayed for hydroxylase activity, receptor translocation, and homology with a rat repetitive DNA sequence. PMID- 3028326 TI - Carcinogenicity testing in nude mouse cell cultures. AB - Secondary cultures of newborn NMRI nu/nu (nude) mouse skin fibroblasts were used as targets for transformation by the combined administration of SV40 and 3 methylcholanthrene (MC). The long (72 h) post-treatment with MC increased virus transformation as much as 4.3-fold. In contrast, anthracene, a non-carcinogenic compound, had no effect on viral transformation frequency. Despite considerable variation within a group, the cell lines transformed by the combination treatment, as a group, were more tumourigenic than cell lines transformed by SV40 alone. PMID- 3028327 TI - Studies on the inhibition of brain synaptosomal Na+/K+-ATPase by mercury chloride and methyl mercury chloride. AB - The effect of mercury chloride (HgCl2) and methyl mercury chloride (MeHg) on brain synaptosomal Na+/K+-ATPase was determined in vitro. It was shown that HgCl2 is a more powerful inhibitor than MeHg. The IC50 were 5 X 10(-7) M for HgCl2 and 2.3 X 10(-6) M for MeHg. A non-competitive type of inhibition was observed with the two mercurials. Removal of contaminating lipids with the non-ionic detergent Lubrol did not affect the inhibition of either mercurial. It is concluded that non-essential lipids do not play a significant role in the inhibitory effect of MeHg and that the potency of these mercurials on to inhibit brain synaptosomal Na+/K+-ATPase largely depends on their capacity to block sulfhydryl groups. PMID- 3028328 TI - Perfused and ventilated guinea-pig lung: a method for evaluating xenobiotic effects on arachidonic acid using formaldehyde. AB - Lipoxygenase as well as cyclooxygenase pathways of arachidonic acid (AA) metabolism are involved in antigen-induced bronchoconstriction in ovalbumin sensitized guinea-pig lungs. Chemical lung challenge induced by aerosol containing formaldehyde (5 ppm up to 20 ppm) was effective in increasing thromboxane (TX) B2 release in a dose-dependent manner, without affecting leukotriene (LT) release. This suggests that xenobiotics acting by non immunologic mechanisms on bronchial mucosa may stimulate the cyclooxygenase pathway of AA metabolism, which, in turn, might cause bronchial and/or lung parenchymal effects of pathophysiological relevance. PMID- 3028330 TI - Effects of diazepam, tofizopam or phenytoin during foetal development on subsequent behaviour and benzodiazepine receptor characteristics in rats. AB - The development of rats was studied for 3 postnatal weeks after prenatal medication with diazepam (10 mg/kg/day), phenytoin (50 mg/kg/day), or tofizopam (50 mg/kg twice daily) given by gastric intubation from day 7 to 21 of pregnancy. The treatments had no effect on the litter size. There were also no differences between the groups in a battery of tests for development (negative geotaxis, righting reflex, cliff avoidance, rotarod, passive avoidance), but the activity spurt seen at postnatal days 18-21 was missing in pups of diazepam-treated mothers. The number of benzodiazepine receptors and their affinity for tritiated flunitrazepam developed similarly in all of the rat groups. Thus, the transient changes in motor development seen 2-3 weeks after birth of rats whose mothers received diazepam during pregnancy do not seem to be related to changes in ontogenesis of benzodiazepine receptors. PMID- 3028329 TI - Effects of methylglyoxal on central and peripheral cholinergic responses. AB - Methylglyoxal (MG) has been shown to have a depolarizing effect on the giant interneurones of the isolated 6th abdominal ganglion of the cock-roach. This effect of MG was inhibited by concentrations of nicotine, d-tubocurarine and physostigmine which blocked transmission at the cholinergic cercal nerve-giant interneurone synapse. MG (5 X 10(-5) to 5 X 10(-4) M) produced concentration dependent contractures of the isolated frog rectus abdominis muscle which were inhibited by d-tubocurarine (10(-4) M) and physostigmine (10(-6) M). MG enhanced the action of acetylcholine and inhibited KCl-evoked contractures whereas it had no effect on the response to carbachol. It is concluded that MG appears to act as a cholinomimetic in both the peripheral and central nervous systems. PMID- 3028331 TI - Pathogenic and structural properties of wild type infectious bursal disease virus (IBDV) and virus grown in vitro. AB - Large plaque (LP) and small plaque (SP) variants of Infectious Bursal Disease Virus (IBDV) which are formed in vitro after serial passages in CE-cells at low or high multiplicities of infection were tested for their pathogenic properties in susceptible chickens. LP virus caused clinical manifestations and destruction of the Bursa of Fabricius (BF) without killing the animals. No signs of a disease appeared after infection with SP virus, and only limited necrotic foci developed in the BF. Coinfection with the SP variant and the highly pathogenic wild type (wt) virus reduced mortality and the severity of the disease. In contrast to the SP variant, which forms incomplete particles of low density, with one or the other of the two genomic RNA segments missing, the two RNA segments characteristic for IBDV were present in approximately equal amounts in wt particles with a buoyant density of 1.29 g/ml isolated from the BF. In spite of an almost total replacement of one of the two major structural polypeptides with a molecular weight of 32 kd by a 28 kd protein, infectivity in this low density fraction was only slightly reduced. This finding underlines the importance of the type of post-translational modification in lymphoid cells or fibroblasts. PMID- 3028332 TI - Association of reduced interleukin-2 production with genetic susceptibility to Pichinde virus in inbred strains of hamsters. AB - Adult inbred MHA hamsters are susceptible to lethal infections with Pichinde virus while inbred LSH hamsters resist such infections. Previous studies demonstrated higher levels of endogenous and induced natural killer (NK) activity in MHA splenocytes than in LSH splenocytes. Preferential replication of Pichinde virus in cells with NK activity was suggested by showing that the greater numbers of infected spleen cells observed in MHA hamsters could be accounted for by a cell population that cosedimented with a peak of NK activity. Increased cellularity of thymi and spleens as well as increased cells sensitive to lymphokines was also found in MHA hamsters as compared to LSH hamsters. In the present study we found that injection of anti-asialo GM 1 serum reduced NK activity but did not alter susceptibility to virus infection. However, MHA hamsters were found to be relatively deficient in the production of interleukin 2 and injection of interleukin 2 altered the mortality of hamsters infected with Pichinde virus. These findings suggest that susceptibility to lethal infection by Pichinde virus is associated with reduced ability to produce interleukin 2 in MHA hamsters. PMID- 3028333 TI - Minimal infective dose of rotavirus. AB - We studied the minimal infective dose of the gastroenteritis virus, rotavirus. Increasingly lower doses [10(4), 10(3), 10(1), 1, 10(-2) plaque forming units (PFU)] of the OSU strain of porcine rotavirus were administered to highly susceptible (colostrum deprived, cesarean derived) newborn miniature swine piglets. In vitro studies showed that virus infectivity was inactivated in piglet gastric juice, both by low pH and by pH- and concentration-dependent factor(s). These factors remain unidentified, but to prevent intragastric viral inactivation, sodium bicarbonate was administered prior to oral virus inoculation of piglets with virulent (non-tissue culture passaged) virus. The lowest dose of virus to induce clinical illness or to demonstrate viral replication by recovery of significantly more infectious virus than was administered, or both, was 1 PFU. These results should help establish standards for virus contamination of water and recommendations for evaluating disinfection procedures for rotaviruses. PMID- 3028335 TI - Comparison of Aujeszky's disease (pseudorabies) virus strains by SDS-PAGE of the virus proteins. AB - Comparison of Aujeszky's disease virus strains by SDS-PAGE indicated that one protein (mol. wt. 31,000-34,000) exhibited marked variation. The NIA 4 and Bartha vaccine strains were identical and could be distinguished from other isolates. PMID- 3028334 TI - Molecular pathogenesis of equine coital exanthema (ECE): temperature sensitivity (TS) and restriction endonuclease (RE) fragment profiles of several field isolates. AB - Examination of six field isolates of equine herpesvirus 3, the causative agent of equine coital exanthema, indicates that all were temperature sensitive (ts) at the body temperature, 39 degrees C, of their host (Equine asinus and callabus) when grown in cell culture. The isolates were characterized by fingerprint analysis with the restriction endonucleases XbaI, EcoRI, BamHI and Hind III to establish possible epidemiologic relatedness. Three of the six isolates may be considered related. Variation in the mobility of the BamHI-A and Hind III-K fragments indicates that a small plaque isolate may contain a 5.7 kb insert of DNA in the unique short region of the genome. PMID- 3028336 TI - In vitro translation of mRNA species from cells infected with Machupo virus. AB - RNA from Machupo virus infected cells was centrifuged in a linear sucrose gradient and RNAs from gradient fractions were tested separately for template activity in a cell-free protein synthesizing system from rabbit reticulocytes. Fraction 15-16 S programmed the synthesis of protein that migrated in SDS polyacrylamide gel like the nucleocapsid protein of a purified virus. The synthesis of virus glycoproteins was not detected in the system. PMID- 3028337 TI - Marked sequence variation between segment 4 genes of human RV-5 and simian SA 11 rotaviruses. AB - The complete nucleotide sequence of dsRNA gene segment 4 of a human serotype 2 rotavirus, RV-5, was determined by sequencing overlapping cloned DNA copies of the gene. Segment 4 is 2359 base pairs in length and contains a single long open reading frame of 2325 bases capable of coding for a protein of 775 amino acids, with 5' and 3' non coding regions of 9 and 25 nucleotides respectively. Comparison with SA 11 segment 4 sequence reveals a moderately conserved trypsin cut site and an overall amino acid homology of 69.8 percent. One localized region of 126 amino acids is only 37.8 percent homologous. Localized frame shifts account for some of this variation, but at the nucleotide level the segment 4 sequences show more variability than other rotavirus genes that have been studied so far. PMID- 3028338 TI - Isolation and serotyping of animal rotaviruses and antigenic comparison with human rotaviruses. Brief report. AB - Rotaviruses were isolated from Australian farm animals (calf, pig and foal) with diarrhoea. Reciprocal cross neutralisation studies showed antigenic similarities with overseas animal strains and with human strains including a newly defined fifth human serotype. PMID- 3028340 TI - Human papillomavirus type 13 DNA in focal epithelial hyperplasia among Mexicans. Brief report. AB - Human papillomavirus (HPV) type 13 DNA was detected in focal epithelial hyperplasia lesions of the oral mucosa in seven half-caste mexicans. The lesions contained intracellular papillomavirus-like particles with a diameter of about 50 nm. DNA extracted from biopsies contained unintegrated HPV type 13 DNA genomes as revealed by Southern blot hybridization. The HPV 13 DNA that was isolated in the present study had the same restriction enzyme cleavage map as HPV 13 DNA, previously described by others. It was moreover confirmed that HPV type 13 genome is related to the genomes of HPV types 6 and 11. PMID- 3028339 TI - Structural and physiological properties of mengovirus: avirulent, hemagglutination-defective mutants express altered alpha (1 D) proteins and are adsorption-defective. AB - Structural and physiological properties of two mutants of mengovirus, 205 and 280, were compared to those of wild-type virus to understand the molecular basis of changes exhibited in their biological function. Two dimensional gel electrophoresis of wild-type and mutant structural proteins revealed alterations in the isoelectric character of the alpha (1 D) protein of both mutant 205 and 280. These data suggest that alterations in the alpha (1 D) protein may be responsible for the phenotypic changes by the mutants. A delay in detectable virus-specified protein synthesis was exhibited in mutant-infected cells in comparison to wild-type. The amount of RNA synthesized in mutant- and revertant infected cells was less than that synthesized in wild-type infected cells. Changes in virus-specified macromolecular synthesis in mutant and revertant infected cells reflected a decrease in the ability of the viruses to attach to cells. PMID- 3028341 TI - Molecular cloning and characterization of Bombyx densovirus genomic DNA. Brief report. AB - A map of the Bombyx densovirus (DNV) genomic DNA has been constructed by cleaving it with several restriction endonucleases. This map shows that this virus and the mammalian parvoviruses are distinct. A restriction fragment containing more than 85 per cent of the complete viral genome has been cloned in a bacterial plasmid. PMID- 3028343 TI - Biological properties of mengovirus: characterization of avirulent, hemagglutination-defective mutants. AB - Biological properties of two mengovirus mutants, 205 and 280, were compared to those of wild-type virus. The mutants exhibited alterations in plaque morphology, hemagglutination, and virulence in mice, but were not temperature-sensitive. Agglutination of human erythrocytes by mengovirus was dependent on the presence of sialic acid on the erythrocyte surface; however, free sialic acid failed to inhibit hemagglutination. Glycophorin, the major sialoglycoprotein of human erythrocyte membranes, exhibited receptor specificity for wild-type virus, but not for mutants 205 or 280. Cross-linking studies indicated that glycophorin exhibited binding specificity for the alpha (1 D) structural protein. The LD50 titers for wild-type mengovirus were 7 and 1500 plaque forming units (PFU) in mice infected intracranially (IC) and intraperitoneally (IP), respectively. However, mice infected IC or IP with 10(6) or 10(7) PFU of mutant 205 or 280 did not exhibit symptoms indicative of virus infection. Revertants were isolated from the brains of mice infected with mutant 205, but not from the brains of mice infected with mutant 280. The biological characterization of the revertants indicated that hemagglutination and virulence may be phenotypically-linked traits. PMID- 3028345 TI - Intraocular pressure assessment in gas-filled eyes following vitrectomy. AB - The accuracy and precision of tonometric methods in measuring intraocular pressure (IOP) was assessed in 24 eye-bank eyes subjected to pars plana lensectomy/vitrectomy and air-fluid exchange. Intraocular pressure was measured in masked fashion with a Perkins' applanation tonometer, pneumatic applanation tonometer; a mercury manometer served as a reference standard. Pneumatic tonometry underestimated actual IOP by as much as 25%, and Schiotz' indentation tonometry underestimated actual IOP by as much as 79%. Perkins' applanation tonometry was significantly more accurate in estimating actual IOP in gas-filled eyes than pneumatic tonometry or Schiotz' indentation tonometry. PMID- 3028344 TI - Cardiovascular-reflex testing and single-fiber electromyography in botulism. A longitudinal study. AB - Four patients with botulism were studied on admission and at different times after intoxication, using a battery of cardiovascular autonomic tests. The results were compared with clinical status and single-fiber electromyographic findings. In the early stage of intoxication, the control of heart-rate and blood pressure responsivity was markedly impaired, as was the neuromuscular transmission. At follow-up, results of sympathetic tests normalized earlier than those of parasympathetic tests. The recovery of autonomic function was slower than that of neuromuscular transmission in three patients. Monitoring autonomic derangement in botulism adds further information on the course of the disease and may identify patients at risk for cardiac or respiratory arrest. Further clinical investigation can help in determining more precisely the autonomic sites where the toxin acts. PMID- 3028342 TI - Evolution of in vitro persistence of two strains of canine parainfluenza virus. Brief report. AB - Lytic stock virus (CPI+) exhibited prominent CPE in Vero cell monolayers. The non syncytial giant cell forming strain (CPI-) was isolated from brain tissues of a dog after experimental infection with syncytial giant cell-forming (CPI+) virus. In mixed infection experiments, CPI(-) virus interfered with the replication of CPI(+) virus in that simultaneously infected cells showed strand-forming CPE typical of CPI(-) and reduction of CPI(+) yield. The CPI(-) virus established immediate in vitro persistence, followed by a severe crisis at passage (p) 8 and several similar crises until p 31; no further crises occurred throughout the remaining observation period (p 80). The CPI(+) did not establish immediate in vitro viral persistence and experienced several lytic crises until p 29. No further crises were observed throughout the observation period (p 100). PMID- 3028346 TI - Experimental tympanosclerosis following infection with Streptococcus pyogenes and vitamin D3 intoxication. AB - A rat animal model was used to study the ultrastructure of submucosal calcifications induced in the middle ear following inoculation with Streptococcus pyogenes and high doses of parenteral vitamin D3. The morphological changes present in affected animals resembled the classical picture of tympanosclerosis. While calcification occurred about bacterial remnants and myelin structures, the most important calcification centers were lysosomal and non-lysosomal matrix vesicles in the extracellular spaces. These formed band-like calcifications close to the basal membrane without affecting the epithelial layer. This animal model offers the possibility of studying the effect of various therapeutic regimens in the treatment of the dynamic tympanosclerotic process. PMID- 3028347 TI - Superoxide initiates oxidation of low density lipoprotein by human monocytes. AB - Human mononuclear cells were used to evaluate the role of superoxide in the oxidation of low density lipoprotein (LDL). Unstimulated cells produced little superoxide or LDL oxidation as assayed by lipid peroxide content. Stimulation of the cells with phorbol myristate acetate (PMA) resulted in an increase both in superoxide production and in LDL oxidation. Mononuclear cell-mediated LDL oxidation was time- and cell number-dependent and was markedly enhanced by the presence of Fe (10 microM). Superoxide was required for the initiation of LDL oxidation as indicated by inhibition of the reaction by early addition of superoxide dismutase (SOD). Propagation of LDL oxidation was superoxide independent, since the later addition of SOD resulted in progressively less inhibition of LDL oxidation. Propagation of LDL oxidation also was, in part, cell independent as indicated by continued oxidation of LDL when mononuclear cells were removed following a 1 to 8 hour period with cells. Optimal LDL oxidation required the presence of mononuclear cells throughout the incubation period, suggesting that cellular factors in addition to superoxide play a role in LDL oxidation. Further evidence for the role of superoxide in the oxidation of LDL by mononuclear cells was obtained with cells from patients with genetic deficiencies of either superoxide generation (chronic granulomatous disease) or myeloperoxidase. PMA-stimulated cells from a patient with chronic granulomatous disease neither generated superoxide nor modified LDL. Incubation of LDL with cells from a patient with myeloperoxidase deficiency (in which superoxide production is normal or increased) resulted in oxidation of the lipoprotein equivalent to that observed with normal cells. Other inhibitors of oxidation reactions also were tested.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3028348 TI - Epstein-Barr virus early antigen induction in nasopharyngeal hybrid cells by Chinese medicinal herbs. AB - The effects of Chinese medicinal herbal drugs (CMH; Daphne genkwa, Wikstroemia indica, Croton oil) were studied for Epstein-Barr virus (EBV) early antigen (EA) induction in established nasopharyngeal hybrid cells. Both ether and water extracts of CMH were found to have inducing activity. However water extracts of the same herbs were not as strong as that of other extracts. The EA positive-cell rate was from 18.2 to 42.2% in ether extracts and 1.0 to 3.8% in water extracts at 10 microgram/ml of the concentration. N-Butyrate alone showed a 40.2% positive rate and in the both treatment of water extracts, a combination effects was seen in induction of the EBV-EA. This in vitro system for the induction of EBV-EA was thought to be useful to determine what is the causal factors for activation of EBV in vivo. PMID- 3028349 TI - Long-term survival in response to combined chemotherapy and radiotherapy in laryngeal small cell carcinoma. AB - Fourteen patients with small cell carcinoma of the larynx are studied. This represents the largest series, from a single institution, reported in the literature. This neoplasm is usually highly aggressive and the prognosis very poor, but, in our experience, combined chemo- and radiotherapy can significantly improve the clinical course of the disease. Three of six patients who received this combined modality treatment are still clinically disease-free more than six years after the initial diagnosis. PMID- 3028350 TI - Effects of infectious bronchitis virus (Arkansas strain) on laying chickens. AB - Seventy-seven-week-old white leghorn layers were inoculated intraocularly with the Arkansas strain of infectious bronchitis virus (AIBV) to study the effects of the virus on egg production and on antibody response of the birds. Infected hens laid fewer eggs than the controls, and those eggs weighed less than eggs laid by controls. Further, the shell quality and internal quality of eggs laid by infected birds were inferior. The serum hemagglutination-inhibition (HI) titers of infected birds increased continuously through 4 weeks postinfection; serum HI titers of the controls were negligible. PMID- 3028351 TI - Effect of maternal antibody on timing of initial vaccination of young white leghorn chickens against infectious bursal disease virus. AB - Maternal antibody titers in white leghorn chicks against infectious bursal disease virus (IBDV) were measured by a computer-assisted, single-serum-dilution, indirect kinetic-based enzyme-linked immunosorbent assay (KELISA) and by a virus neutralization (VN) test in order to predict the timing of initial vaccination. Day-old white leghorns were from unvaccinated pullets or from pullets vaccinated either four times or twice with IBDV commercial vaccines. The chicks were immunized once via the drinking water with a commercial "intermediate" live IBDV vaccine at 1, 15, or 28 days of age. Effective initial immunization was confirmed by an increase in antibody to IBDV (serologic conversion) that occurred when maternal antibody decreased to 8 and 9 on a log2 scale. This concentration of antibody was detected between 24 and 28 days of age. The computer-assisted IBDV KELISA increased the sample processing speed for detecting IBDV antibody, and it was as sensitive as the VN test for predicting the timing of initial IBDV vaccination. PMID- 3028352 TI - Quantitation of hemorrhagic enteritis virus antigen and antibody using enzyme linked immunosorbent assays. AB - Enzyme-linked immunosorbent assays (ELISAs) were developed to quantitate hemorrhagic enteritis virus (HEV) antibodies in turkey sera and HEV antigens in tissue extracts. These assays were more sensitive than the commonly used agar-gel precipitin tests in detecting antigen and antibody. The antibody-ELISA was used to monitor the presence and decline of passive antibodies in turkey poults and the seroconversion of turkeys infected with HEV. The antigen-ELISA was carried out using a monoclonal antibody; this test was used to quantitate HEV antigen in experimentally infected turkeys in a time-sequence experiment. Both ELISAs measured a strong antigenic relationship between an avirulent strain (HEV-A) and a virulent strain (HEV-V). PMID- 3028353 TI - Influence of dietary calcium stress on lethality of avian influenza viruses for laying chickens. AB - The effect of calcium stress was studied in an attempt to reproduce lethal infections in laying chickens with A/Chicken/Alabama/75 (H4N8) influenza virus and with two nonpathogenic H5N2 influenza viruses from the 1983-84 outbreak in the eastern United States. Hens were fed calcium-deficient or standard diets for 7 to 14 days; then the calcium-deficient feed was replaced with standard feed supplemented with ad libitum oyster shell, and both groups of hens were inoculated with virus. When hens were infected with the H4N8 virus, respective mortalities of those on the calcium-deficient and standard diets were 19% (27/141) and 5% (7/143). The H5N2 viruses did not kill hens fed either diet. In standard pathogenicity tests, Alabama H4N8 viruses reisolated from the hens that died generally were more lethal for 4-week-old chickens than the stock virus. These results argue for characterization of the Alabama H4N8 virus as pathogenic rather than nonpathogenic as originally determined. PMID- 3028355 TI - Histopathologic changes induced by serotype II infectious bursal disease virus in specific-pathogen-free chickens. AB - Specific-pathogen-free (SPF) chickens were infected with infectious bursal disease virus (IBDV) serotype II (turkey, Missouri isolate) at 1 day or 4 weeks of age. Chickens infected at 1 day had small bursas 3 weeks postinfection (PI). Bursas were necrotic and inflamed through 1 week PI. Post-necrotic atrophy of the bursas was marked at 3 weeks PI, and plasma cells in the Harderian gland were depleted. Chickens infected at 4 weeks had slightly enlarged spleens with small gray foci on the surface at 1 week PI. The bursas had massive necrosis and hemorrhages on the mucosal surface and had gelatinous yellowish transudate covering the serosal surface; some bursas were gray. Bursas were smaller than normal through the end of the experiment (4 1/2 weeks PI). At 1 week PI, histologic lesions were multifocal lymphoid necrosis of spleens and necrotic inflammation of the bursas. Post-necrotic atrophy of the bursas was found 2 weeks PI through the end of the experiment. Compared with uninfected controls, infected chickens had significantly more pyroninophilic blast cells in the spleen at 1 week PI and the bursas at 4 1/2 weeks PI. Plasma cells in the Harderian glands of this infected group were depleted as well. PMID- 3028354 TI - Standardization and application of the enzyme-linked immunosorbent assay for infectious bronchitis. AB - The indirect enzyme-linked immunosorbent assay (ELISA) was used to detect antibody to infectious bronchitis virus in chickens. The serum-neutralization test for infectious bronchitis (SNIB) was used as a reference serologic test. The ELISA proved to be useful for monitoring antibody responses following vaccination of leghorn chicks. The titers obtained with the ELISA and SNIB showed an overall correlation, but results suggest the involvement of different antibody classes. PMID- 3028356 TI - Serological profiles of commercial broiler breeders and their progeny. 1. Infectious bronchitis virus. AB - Serum samples were collected from broiler breeders and their 1-day-old, 2-week old, and 5-week-old progeny from different regions of the United States. Individual samples were tested by hemagglutination-inhibition (HI) against six infectious bronchitis virus (IBV) strains: Massachusetts 41 (Mass), H52, Connecticut 46 (Conn), Arkansas 99, SE17, and JMK. The use of multiple strains to test broiler flocks resulted in the detection of seroconversions to Conn and JMK vaccination that were not detected with the IBV Mass HI test. Further, HI titers were detected to IBV strains not used for flock vaccination. In some cases, those titers could be due to cross reactions to antigens common to each of the virus strains. In two breeder flocks, the highest HI titers were to heterologous strains. PMID- 3028357 TI - Embryo vaccination of specific-pathogen-free chickens with infectious bursal disease virus: tissue distribution of the vaccine virus and protection of hatched chickens against disease. AB - Vaccination of specific-pathogen-free chickens as 18-day embryos with the BVM isolate of infectious bursal disease virus (IBDV) resulted in extensive replication of the vaccine virus in the embryonic tissues. The virus was recovered from lung, thymus, proventriculus, liver, kidney, and spleen of embryos 1 day postvaccination, and recoverable virus persisted for at least 7 days. Replication and spread of the vaccine virus in chickens vaccinated as 18-day embryos was compared with that in chickens vaccinated at hatch. Distribution of the virus in tissues was more extensive, virus levels in tissues were generally higher, and detectable virus persisted longer in chickens vaccinated as 18-day embryos than in those vaccinated at hatch. Effective vaccine response could be initiated with 6.2 median embryo lethal doses, the lowest dose tested. Chickens immunized as embryos developed neutralizing antibody against IBDV and resisted challenge with pathogenic IBDV at 4, 6, 8, and 10 weeks of age. PMID- 3028358 TI - Electropherotypic analysis of rotaviruses isolated from turkeys. AB - From 1980 to 1984, several flocks of turkeys in Minnesota exhibiting signs of clinical enteritis were examined for viruses. Electron microscopic (EM) examination of fecal specimens from 35 flocks revealed the presence of rotavirus particles. Rotaviruses were successfully isolated in cell cultures from only 24 of these positive fecal specimens. Double-stranded RNA (dsRNA) preparations made from these 24 cell-culture isolates and from the remaining 11 fecal samples that were rotavirus-positive on EM examination were analyzed by polyacrylamide gel electrophoresis for genetic differences in their genomes. The study revealed eight distinct electropherotypes among the rotavirus dsRNA preparations. Atypical dsRNA migration patterns were recognized only in preparations of dsRNA from fecal materials. PMID- 3028359 TI - Genomic variation among avian rotavirus-like viruses detected by polyacrylamide gel electrophoresis. AB - The genomes of five turkey and two chicken rotavirus-like virus (RVLV) strains were compared with the genome of a reference turkey RVLV strain by co electrophoresis in polyacrylamide gels. The genomes of these avian RVLV strains could be distinguished from the reference strain genome by differences in the mobilities of from three to 10 segments. PMID- 3028360 TI - Restriction endonuclease patterns of some European and American isolates of avian infectious laryngotracheitis virus. AB - Eleven isolates of infectious laryngotracheitis virus (nine European and two American) were compared by restriction endonuclease analysis of their DNA after radiolabeling with 32P. Digestion with KpnI gave identical cleavage patterns for all the European isolates, but the two American viruses (one field and one vaccine) showed some differences from them and from each other. In the case of the American vaccine strain, however, these differences were only minor. After BamHI digestion, only the American field isolate appeared to be different, whereas with HindIII, all 11 isolates were identical. PMID- 3028362 TI - Topical interferon cream for the treatment of herpes genitalis: a double-blind controlled trial. AB - Thirty-five women with primary herpes genitalis and 34 women with recurrent herpes genitalis were treated with either placebo, or beta Interferon cream (20,000 iu/g). In comparison with the placebo group, the treatment group showed no benefit in either symptom relief, speed of healing of lesions or length of viral shedding time from lesions. PMID- 3028361 TI - Laryngotracheitis outbreak limited to a part of a chicken flock exposed to smoke and chemicals. AB - Laryngotracheitis was diagnosed in a flock of molted, caged table-egg-layers. Morbidity was restricted to an area of the house in which the birds had been exposed to smoke from a fire in the house and to a powdered chemical fire extinguisher used on the fire. Mortality in this group began to rise 6 days after the fire and continued to be above normal for about 3 weeks. Feed consumption dropped for about 1 1/2 weeks after the fire but was normal or above normal during the rest of the disease outbreak. Egg production dropped slightly for 1 week after the fire, then returned to normal. Microscopic tracheitis in the exposed birds continued for 11 weeks after the fire. Hens outside of the smoke affected area did not show histopathological changes or shed laryngotracheitis virus. PMID- 3028363 TI - Alcohol drinking patterns of young adult rats as a function of infantile aversive experiences with alcohol odor. AB - Recent studies have demonstrated that in the rat, alcohol intake patterns can be regulated by prior experiences with the odor of this drug. The efficacy of such regulation appeared to be limited to early postnatal stages of development. The present study supports the possibility, however, that early ethanol odor experiences are retained over considerable amounts of time and may play an effective role in the control of adult patterns of alcohol ingestion. Specifically, it was observed that rats exposed to pairings of ethanol odor and lithium chloride toxicosis (Etoh-LiCl group) during Postnatal Days 5, 10, 15, and 20 displayed as young adults (52-58 days) significant decreases in voluntary alcohol intake scores. This decrease was determined relative to control rats that had been safely exposed to ethanol odor (Etoh group) or given only lithium toxicosis but no ethanol odor (LiCl group) during Postnatal Days 5, 10, 15, and 20. Relative to these controls (Etoh and LiCl groups) the experimental subjects (Etoh-LiCl group) also exhibited significant decreases in their adult preference for ethanol odor, as assessed through an olfactory locational test. The present results indicate several ways in which conditioned aversion to ethanol intake may arise and imply that the transfer between olfactory and gustatory aversions takes place at the time of memory storage rather than at some later stage of memory processing. In a more general sense, the present results add to others in our series to support the notion that consideration of the effects of early experience with alcohol may aid in the clarification and control of voluntary alcohol intake patterns. PMID- 3028364 TI - Importance of tactile and olfactory cues to the inhibition of the grasshopper mouse's attack through toxicosis. AB - This study examined the effect that toxicosis paired with the presence of a distinctive texture had on the inhibition of the grasshopper mouse's predatory attack. The first experiment measured the context in which tactile cues would be most effective by presenting prey with various combinations of added stimuli. The combination of distinctive tactile, olfactory, and gustatory cues produced the longest lasting inhibition. Because certain manipulations also had an unintended weak odor associated with them, the second experiment measured the importance of a weak or strong odor to inhibition of a mouse's attack. A combination of strong odor and distinctive texture paired with toxicosis inhibited an attack more effectively than a weak odor and a similar texture. In a third experiment, toxicosis was paired with an acraea moth caterpillar which has a highly distinctive texture. This produced the longest lasting inhibition of mouse's attack observed thus far. The ecological significance of a combination of a distinctive texture and odor to the inhibition of the grasshopper mouse's attack is discussed. PMID- 3028365 TI - Odor-aversion learning and retention span in neonatal mouse pups. AB - One hundred and sixty-four litters of Swiss CD-1 random-bred mice were used to assess learning and retention capacities during the first postnatal week. In Experiment 1, whole 7-day litters were exposed for 65 min to commercial extracts of either mint or lemon sprinkled over wood shavings. Five minutes after the beginning of the exposure, half of the litters were injected ip with the illness inducing agent lithium chloride (LiCl; 0.20 M, 2% of body weight); the other half was treated with saline solution (8% NaCl). On Postnatal Day 10, the animals were singly introduced in a warmed arena for a 180-s preference test, and the time spent in the mint- and lemon-scented areas of the apparatus was recorded. When compared with saline-injected pups, mice that experienced lemon-LiCl pairings showed a significant aversion for the lemon-scented area, while the mint aversion in the mint-LiCl group just missed statistical significance. Three additional control groups (unhandled on Day 7, or only LiCl- or saline-injected) did not show significant preferences for either the mint or the lemon odor. In Experiment 2, litters of 3, 5, or 7 days were similarly exposed to lemon-scented shavings for either 5 or 20 min, injected with LiCl or saline, and then exposed for an additional 60 min to the shavings. On Postnatal Day 10, tests like those of Experiment 1 showed a significant odor-aversion in animals conditioned on Day 7, but not in those conditioned on Day 3 or 5. In Experiment 3, 3- and 5-day old pups were exposed to lemon odor-LiCl or -NaCl pairings, and tested for aversion after 3 or 7 days (CS duration 5 min before injection and either 30 or 60 min after injection). Only when the conditioning-testing interval was limited to 3 days did LiCl-injected groups show a significant aversion, which did not depend on duration of CS exposure. PMID- 3028366 TI - Neonatal hyperthyroidism in the rat: thyroxine accelerates the development of unconditioned but not learned responses to tastes. AB - The effects of neonatal hyperthyroidism induced by thyroxine injection on the development of unconditioned and learned behaviors mediated by the gustatory system of the rat were investigated. The development of unconditioned ingestive responses evoked by 10% sucrose and 0.1% HCl taste solutions was advanced several days by thyroxine treatment. However, the emergence of taste aversion learning involving 10% sucrose and LiCl injection was not advanced and may have been slightly delayed. Thus, the ontogenesis of unconditioned and learned behaviors mediated by the gustatory system was not influenced uniformly by neonatal thyroxine treatment. PMID- 3028367 TI - Secretin induces rapid increases in inositol trisphosphate, cytosolic Ca2+ and diacylglycerol as well as cyclic AMP in rat pancreatic acini. AB - Previous studies have shown that the dose-response relationship for secretin stimulated cyclic AMP accumulation is different from that for secretin-stimulated enzyme secretion in the rat exocrine pancreas. Here we show that secretin concentrations of 10(-10) M and higher stimulated a rise in cyclic AMP levels, with maximum effect on cyclic AMP accumulation being achieved already with 10(-8) M-secretin. However, at this concentration of secretin, enzyme secretion rates were approximately half-maximal. Unexpectedly, at concentrations of secretin greater than 10(-8) M there was evidence suggestive of phosphatidylinositol bisphosphate hydrolysis with rapid increases in inositol trisphosphate, cytosolic free calcium and diacylglycerol content of rat pancreatic acini. Furthermore, there was a dose-response relationship among secretin concentration (in the range 10(-8) M-2 X 10(-6) M), increases in inositol trisphosphate and increases in cytosolic free calcium ([Ca2+]i). Contrary to what has been previously believed, these results clearly indicate that in rat pancreatic acini secretin not only stimulates cyclic AMP accumulation but also raises inositol trisphosphate, [Ca2+]i and diacylglycerol. Thus, two second messenger systems may play a role in the regulation of secretin-induced amylase release. PMID- 3028369 TI - Charged anaesthetics alter LM-fibroblast plasma-membrane enzymes by selective fluidization of inner or outer membrane leaflets. AB - The functional consequences of the differences in lipid composition and structure between the two leaflets of the plasma membrane were investigated. Fluorescence of 1,6-diphenylhexa-1,3,5-triene(DPH), quenching, and differential polarized phase fluorimetry demonstrated selective fluidization by local anaesthetics of individual leaflets in isolated LM-cell plasma membranes. As measured by decreased limiting anisotropy of DPH fluorescence, cationic (prilocaine) and anionic (phenobarbital and pentobarbital) amphipaths preferentially fluidized the cytofacial and exofacial leaflets respectively. Unlike prilocaine, procaine, also a cation, fluidized both leaflets of these membranes equally. Pentobarbital stimulated 5'-nucleotidase between 0.1 and 5 mM and inhibited at higher concentrations, whereas phenobarbital only inhibited, at higher concentrations. Cationic drugs were ineffective. Two maxima of (Na+ + K+)-ATPase activation were obtained with both anionic drugs. Only one activation maximum was obtained with both cationic drugs. The maximum in activity below 1 mM for all four drugs clustered about a single limiting anisotropy value in the cytofacial leaflet, whereas there was no correlation between activity and limiting anisotropy in the exofacial leaflets. Therefore, although phenobarbital and pentobarbital below 1 mM fluidized the exofacial leaflet more than the cytofacial leaflet, the smaller fluidization in the cytofacial leaflet was functionally significant for (Na+ + K+)-ATPase. Mg2+-ATPase was stimulated at 1 mM-phenobarbital, unaffected by pentobarbital and slightly stimulated by both cationic drugs at concentrations fluidizing both leaflets. Thus the activity of (Na+ + K+)-ATPase was highly sensitive to selective fluidization of the leaflet containing its active site, whereas the other enzymes examined were little affected by fluidization of either leaflet. PMID- 3028368 TI - Adipose-tissue Mg2+-dependent phosphatidate phosphohydrolase. Control of activity and subcellular distribution in vitro and in vivo. AB - The subcellular distribution of Mg2+-dependent phosphatidate phosphohydrolase in rat adipocytes between a soluble and a membrane-bound fraction was measured by using both centrifugal fractionation and a novel Millipore-filtration method. The relative proportion of the phosphohydrolase associated with the particulate fraction was increased on incubation of cells with noradrenaline or palmitate. Insulin on its own decreased the proportion of the phosphohydrolase that was particulate and abolished the effect of noradrenaline, but not that of palmitate. The effect of noradrenaline on phosphohydrolase distribution was rapid, the effect being maximal within 10 min. Noradrenaline exerted this effect with a similar concentration-dependence to its lipolytic effect. Inclusion of albumin in homogenization buffers decreased the proportion of the phosphohydrolase that was particulate, but did not abolish the effect of noradrenaline. There was limited correlation between the proportion of the phosphohydrolase that was particulate and the measured rate of triacylglycerol synthesis in adipocytes incubated under a variety of conditions. Starvation, streptozotocin-diabetes and hypothyroidism decreased the specific activities of the phosphohydrolase and glycerolphosphate acyltransferase in homogenates from epididymal fat-pads. Restoration of these activities in the diabetic state was seen after administration of insulin over 2 days or, in the short term, within 2 h after a single administration of insulin. Administration of thyroxine over 3 days caused restoration of these activities in the hypothyroid state. Starvation and diabetes increased the proportion of the phosphohydrolase found in the microsomal fraction. This change was not seen when albumin was present in homogenization buffers. The possible role of fatty acids as regulators of the intracellular translocation of the phosphohydrolase, together with the role of this enzyme in the regulation of triacylglycerol synthesis in adipose tissue, is discussed. PMID- 3028370 TI - Inhibition of prolyl 4-hydroxylase by hydroxyanthraquinones. AB - Prolyl 4-hydroxylase (EC 1.14.11.2) is an essential enzyme in the post translational modification of collagen. Inhibitors of this enzyme are of potential interest for the treatment of diseases involving excessive deposition of collagen. We have found that anthraquinones with at least two hydroxy groups ortho to each other are potent inhibitors of this enzyme. Kinetic studies revealed that 2,7,8-trihydroxyanthraquinone (THA) competitively inhibited the co substrate, 2-oxoglutarate, but was non-competitive with regard to ascorbate and was tentatively considered to be uncompetitive with regard to protocollagen. The inhibition by THA was greatly enhanced in the absence of added Fe2+ and was partially reversed by the addition of concentrations of Fe2+ in excess of the optimum for the enzymic reaction. Binding studies indicated that THA is an effective chelating agent for Fe2+. Several non-quinoidal compounds bearing the catechol moiety also inhibited the enzyme. The results suggest that THA inhibited prolyl 4-hydroxylase by binding to the enzyme at the site for 2-oxoglutarate possibly involving the Fe2+ atom, rather than by complexing with Fe2+ in free solution. The inhibition of prolyl 4-hydroxylase by THA exhibited strong positive co-operativity and may involve three distinct but non-independent binding sites. PMID- 3028371 TI - A new non-covalent complex of semisynthetically modified tryptic fragments of cytochrome c. AB - We have prepared a semisynthetic analogue of fully acetimidylated horse cytochrome c, a complex in which the peptide bond between residues glycine-37 and arginine-38 is lacking. In contrast with the complex that we have previously described [Harris & Offord (1977) Biochem. J. 161, 12-25], in which the break in continuity is between residues arginine-38 and lysine-39, the new analogue has a nearly normal redox potential, and can more fully restore succinate oxidation to mitochondria depleted of cytochrome c. Studies of this and other analogues lead us to propose an explanation for the low biological activity of complex (1-38) (39-104) and a role for the invariance of arginine-38. PMID- 3028372 TI - Steroidogenic effect of atrial natriuretic factor in isolated mouse Leydig cells is mediated by cyclic GMP. AB - The effects of different atrial natriuretic peptides on cyclic GMP formation and steroidogenesis have been studied in Percoll-purified mouse Leydig cells. Rat atrial peptides rANP (rat atrial natriuretic peptide), rAP-I (rat atriopeptin I) and rAP-II (rat atriopeptin II), in the presence of a phosphodiesterase inhibitor, stimulated cyclic GMP formation in a concentration-dependent manner. In the presence of saturating concentrations of the peptides, a 400-600 fold stimulation of cyclic GMP accumulation was observed. Among the peptides, rAP-II appeared to be the most potent. ED50 values (concentration causing half-maximal effect) for rAP-II, rANP and rAP-I were 1 X 10(-9) M, 2 X 10(-9) M and 2 X 10(-8) M respectively. A parallel stimulation of cyclic GMP formation and testosterone production by the cells was observed after incubation of the cells with various concentrations of rAP-II. In the presence of a saturating concentration of rAP-II (2 X 10(-8) M), maximum stimulation of intracellular cyclic GMP content was obtained within 5 min of incubation. Testosterone production by mouse Leydig cells could be stimulated by 8-bromo cyclic GMP in a concentration-related manner. At a 10 mM concentration of the cyclic nucleotide, steroidogenesis was stimulated to a similar extent as that obtained with a saturating concentration of human chorionic gonadotrophin (5 ng/ml). On the basis of these results we conclude that cyclic GMP acts as a second messenger in atrial-peptide-stimulated steroidogenesis in mouse Leydig cells. The steroidogenic effect of atrial peptides appears to be species-specific, since none of these peptides stimulated testosterone production by purified Leydig cells of rats, though in these cells a 40-60-fold stimulation of cyclic GMP formation in response to each of the three peptides was observed. However, 8-bromo cyclic GMP could stimulate testosterone production in rat Leydig cells. Therefore we conclude that the lack of steroidogenic response in rat Leydig cells to atrial-natriuretic-factor stimulation results from an insufficient formation of cyclic GMP in these cells. This species difference would appear to result from a lower guanylate cyclase activity in rat Leydig cells. PMID- 3028373 TI - Leukotriene B4 stimulation of phagocytes results in the formation of inositol 1,4,5-trisphosphate. A second messenger for Ca2+ mobilization. AB - Inositol trisphosphate (InsP3) production and cytosolic free Ca2+ ([Ca2+]i) elevations induced by leukotriene B4 (LTB4)-receptor activation were studied in the human promyelocytic-leukaemia cell line HL60, induced to differentiate by retinoic acid. The myeloid-differentiated HL60 cells respond to LTB4 by raising their [Ca2+]i with a dose-response relationship similar to that shown by normal human neutrophils. The observations of the LTB4 transduction mechanism were compared with those of the transduction mechanism of the chemotactic peptide fMet Leu-Phe in HL60 cells differentiated with dimethyl sulphoxide. Both LTB4 and fMet Leu-Phe triggered a rapid (less than 5 s) elevation of [Ca2+]i, which occurred in parallel with the InsP3 production from myo-[3H]inositol-labelled cells. The threshold concentrations of the agonists, for InsP3 production, were found at 10( 9) M, a slightly higher concentration than that required to detect [Ca2+]i elevations. No significant changes were noted in the phosphoinositide levels upon stimulation with LTB4. Exposure to Bordetella pertussis toxin before LTB4 stimulation abolished both the increased formation of InsP3 and the rise of [Ca2+]i. LTB4 and fMet-Leu-Phe elicited elevations of inositol 1,4,5 trisphosphate [Ins(1,4,5)P3] with no detectable lag time, followed by slower and more sustained inositol 1,3,4-trisphosphate elevations. Stimulation with various leukotriene analogues revealed a good correlation between both total InsP3 as well as Ins(1,4,5)P3 formation and elevations of [Ca2+]1. Thus LTB4 receptor activation results in an increased production of Ins(1,4,5)P3 via a transduction mechanism also involving a nucleotide regulatory protein, as previously described for the fMet-Leu-Phe transduction mechanism. PMID- 3028375 TI - Biosynthesis of gamma-linolenic acid in cotyledons and microsomal preparations of the developing seeds of common borage (Borago officinalis). AB - The developing seeds of Borago officinalis (common borage) accumulate a triacylglycerol oil that is relatively rich in the uncommon fatty acid gamma linolenate (octadec-6,9,12-trienoic acid). Incubation of developing, whole, cotyledons with [14C]oleate and [14C]linoleate showed that the gamma-linolenate was synthesized by the sequential desaturation of oleate----linoleate----gamma linolenate. Microsomal membrane preparations from the developing cotyledons contained an active delta 6-desaturase enzyme that catalysed the conversion of linoleate into gamma-linolenate. Experiments were designed to manipulate the [14C]linoleate content of the microsomal phosphatidylcholine. The [14C]linoleoyl phosphatidylcholine labelled in situ was converted into gamma-linolenoyl phosphatidylcholine in the presence of NADH. The substrate for the delta 6 desaturase in borage was, therefore, the linoleate in the complex microsomal lipid phosphatidylcholine, rather than, as in animals, the acyl-CoA. This was further confirmed in experiments that compared the specific radioactivity of the gamma-linolenate, in acyl-CoA and phosphatidylcholine, that was synthesized when [14C]linoleoyl-CoA was incubated with microsomal membranes, NADH and non radioactive gamma-linolenoyl-CoA. The delta 6-desaturase was positionally specific and only utilized the linoleate in position 2 of sn-phosphatidylcholine. Analysis of the positional distribution of fatty acids in the endogenous microsomal sn-phosphatidylcholine showed that, whereas position 1 contained substantial linoleate, only small amounts of gamma-linolenate were present. The results shed further light on the synthesis of C18 polyunsaturated fatty acids in plants and in particular its relationship to the regulation of the acyl quality of the triacylglycerols in oilseeds. PMID- 3028374 TI - Differential regulation by phosphatidylinositol 4,5-bisphosphate of pituitary plasma-membrane and cytosolic phosphoinositide kinases. AB - Regulation of phosphatidylinositol kinase (EC 2.7.1.67) and phosphatidylinositol 4-phosphate (PtdIns4P) kinase (EC 2.7.1.68) was investigated in highly enriched plasma-membrane and cytosolic fractions derived from cloned rat pituitary (GH3) cells. In plasma membranes, phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2] added exogenously enhanced incorporation of [32P]phosphate from [gamma-32P]MgATP2 into PtdIns(4,5)P2 and PtdIns4P to 150% of control; half-maximal effect occurred with 0.03 mM exogenous PtdIns(4,5)P2. Exogenous PtdIns4P and phosphatidylinositol (PtdIns) had no effect. When plasma membranes prepared from cells prelabelled to isotopic steady state with [3H]inositol were used, there was a MgATP2- dependent increase in the content of [3H]PtdIns(4,5)P2 and [3H]PtdIns4P that was enhanced specifically by exogenous PtdIns(4,5)P2 also. Degradation of 32P- and 3H-labelled PtdIns(4,5)P2 and PtdIns4P within the plasma-membrane fraction was not affected by exogenous PtdIns(4,5)P2. Phosphoinositide kinase activities in the cytosolic fraction were assayed by using exogenous substrates. Phosphoinositide kinase activities in cytosol were inhibited by exogenously added PtdIns(4,5)P2. These findings demonstrate that exogenously added PtdIns(4,5)P2 enhances phosphoinositide kinase activities (and formation of polyphosphoinositides) in plasma membranes, but decreases these kinase activities in cytosol derived from GH3 cells. These data suggest that flux of PtdIns to PtdIns4P to PtdIns(4,5)P2 in the plasma membrane cannot be increased simply by release of membrane-associated phosphoinositide kinases from product inhibition as PtdIns(4,5)P2 is hydrolysed. PMID- 3028376 TI - The dependence on Ca2+ of the guanine-nucleotide-activated polyphosphoinositide phosphodiesterase in neutrophil plasma membranes. AB - The requirement for Ca2+ for the activation of polyphosphoinositide phosphodiesterase was studied with the guanine nucleotide analogue guanosine 5' [gamma-thio]triphosphate (GTP gamma S). Levels of Ca2+ that pertain in unstimulated neutrophils (100 nM) are obligatory for the full expression of enzyme activity stimulated with GTP gamma S. Reduction of Ca2+ to 1 nM leads to inhibition. Increasing the level of Ca2+ from 100 nM to 1000 nM does not alter enzyme activity. Guanosine 5'-[beta-thio]diphosphate (GDP beta S) does not stimulate the phosphodiesterase but is an effective inhibitor of activation by GTP gamma S. Ca2+ in the millimolar range can also activate the phosphodiesterase alone and this is not inhibited by GDP beta S. It is also shown that Sr2+ in the millimolar range can stimulate enzyme activity similarly to Ca2+. PMID- 3028379 TI - Assay of prolyl 4-hydroxylase by the chromatographic determination of [14C]succinic acid on ion-exchange minicolumns. AB - An assay for prolyl 4-hydroxylase (EC 1.14.11.2) is described which measures succinic acid produced during the decarboxylation of 2-oxoglutaric acid in the presence of poly(L-Pro-Gly-L-Pro). [1-14C]Succinic acid was separated from its precursor 2-oxo[5-14C]glutaric acid by using ion-exchange minicolumns. The contamination of succinic acid by 2-oxoglutaric acid was approx. 1%, and the recovery of succinic acid was 100%. Kinetic parameters of prolyl 4-hydroxylase measured by the assay showed good agreement with published values. Our experience indicates that the measurement of prolyl 4-hydroxylase by the production of succinic acid is especially suited to investigations involving large numbers of assays. PMID- 3028380 TI - Lithium suppresses seizure susceptibility in pentylenetetrazol-kindled rats. AB - With regard to the still debatable effect of lithium in epilepsy the effect of chronic lithium treatment on the development of pentylenetetrazol(PTZ)-kindling and the anti-convulsive potency of acute lithium administration on PTZ-induced seizure behaviour in PTZ-kindled and non-kindled rats was examined. Lithium significantly suppressed the development of PTZ-kindling. In kindled rats the acute application of lithium reduced seizure susceptibility in a dose-dependent manner. In contrast to kindled rats, in non-kindled animals lithium had very weak anticonvulsive potency. The results suggest that lithium may be of potential interest in the treatment of special forms of epilepsy and epileptic brain disorders. PMID- 3028377 TI - Conversion of human placental alkaline phosphatase from a high Mr form to a low Mr form during butanol extraction. An investigation of the role of endogenous phosphoinositide-specific phospholipases. AB - Alkaline phosphatase in a wide range of tissues has been shown to be anchored in the membrane by a specific interaction with the polar head group of phosphatidylinositol. It has previously been suggested that the production of low Mr alkaline phosphatase during the commonly used butanol extraction procedure may result from the activation of an endogenous phosphoinositide-specific phospholipase C which removes the 1,2-diacylglycerol responsible for membrane anchoring. This conversion process was investigated in greater detail with human placenta used as the source of alkaline phosphatase. Mr and hydrophobicity of the alkaline phosphatase were determined by gel filtration on TSK-250 and partitioning in Triton X-114, respectively. Alkaline phosphatase extracted from human placental particulate fraction with butanol at pH 5.4 or released by incubation with Staphylococcus aureus phosphatidylinositol-specific phospholipase C produced a form of alkaline phosphatase of Mr approx. 170,000 and relatively low hydrophobicity. By contrast, the butanol extract prepared at pH 8.3 was an aggregated form of Mr approx. 600,000 and was relatively hydrophobic. The effect of a variety of inhibitors and activators on the amount of low Mr alkaline phosphatase produced during butanol extraction revealed that it was a Ca2+- and thiol-dependent process. Proteinase inhibitors had no effect. [3H]Phosphatidylinositol hydrolysis by the particulate fraction, unlike low Mr alkaline phosphatase production, was relatively sensitive to heat inactivation, indicating that the phosphoinositide-specific phospholipases C from cytosol and lysosomes were unlikely to be responsible for conversion. A butanol-stimulated activity which removed the [3H]myristic acid from the variant surface glycoprotein ( [3H]mfVSG) of Trypanosoma brucei was detectable in the human placental particulate fraction. Since this activity was acid active, Ca2+- and thiol-dependent and relatively heat stable, it may be the same as that responsible for production of low Mr alkaline phosphatase. The only 3H-labelled product identified was phosphatidic acid, suggesting that the [3H]mfVSG-cleaving activity is a phospholipase D. These data strongly support the proposal that production of low Mr alkaline phosphatase during butanol extraction is an autolytic process occurring as the result of an endogenous phospholipase. However, they also suggest that the lysosomal and cytosolic phosphoinositide specific phospholipases C that have previously been described in many mammalian tissues are not responsible for this process. PMID- 3028378 TI - Use of forskolin to study the relationship between cyclic AMP formation and bone resorption in vitro. AB - The effect of the adenylate cyclase activator forskolin on bone resorption and cyclic AMP accumulation was studied in an organ-culture system by using calvarial bones from 6-7-day-old mice. Forskolin caused a rapid and fully reversible increase of cyclic AMP, which was maximal after 20-30 min. The phosphodiesterase inhibitor rolipram (30 mumol/l), enhanced the cyclic AMP response to forskolin (50 mumol/l) from a net cyclic AMP response of 1234 +/- 154 pmol/bone to 2854 +/- 193 pmol/bone (mean +/- S.E.M., n = 4). The cyclic AMP level in bones treated with forskolin (30 mumol/l) was significantly increased after 24 h of culture. Forskolin, at and above 0.3 mumol/l, in the absence and the presence of rolipram (30 mumol/l), caused a dose-dependent cyclic AMP accumulation with an calculated EC50 (concentration producing half-maximal stimulation) value at 8.3 mumol/l. In 24 h cultures forskolin inhibited spontaneous and PTH (parathyroid hormone) stimulated 45Ca release with calculated IC50 (concentration producing half maximal inhibition) values at 1.6 and 0.6 mumol/l respectively. Forskolin significantly inhibited the release of 3H from [3H]proline-labelled bones stimulated by PTH (10 nmol/l). The inhibitory effect by forskolin on PTH stimulated 45Ca release was significant already after 3 h of culture. In 24 h cultures forskolin (3 mumol/l) significantly inhibited 45Ca release also from bones stimulated by prostaglandin E2 (1 mumol/l) and 1 alpha hydroxycholecalciferol (0.1 mumol/l). The inhibitory effect of forskolin on spontaneous and PTH-stimulated 45Ca release was transient. A dose-dependent stimulation of basal 45Ca release was seen in 120 h cultures, at and above 3 nmol of forskolin/l, with a calculated EC50 value at 16 nmol/l. The stimulatory effect of forskolin (1 mumol/l) could be inhibited by calcitonin (0.1 unit/ml), but was insensitive to indomethacin (1 mumol/l). Forskolin increased the release of 3H from [3H]proline-labelled bones cultured for 120 h and decreased the amount of hydroxyproline in bones after culture. Forskolin inhibited PTH-stimulated release of Ca2+, Pi, beta-glucuronidase and beta-N-acetylglucosaminidase in 24 h cultures. In 120 h cultures forskolin stimulated the basal release of minerals and lysosomal enzymes.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3028382 TI - Low-spin sulfite reductases: a new homologous group of non-heme iron-siroheme proteins in anaerobic bacteria. AB - Two new low molecular weight proteins with sulfite reductase activity, isolated from Methanosarcina barkeri (DSM 800) and Desulfuromonas acetoxidans (strain 5071), were studied by EPR and optical spectroscopic techniques. Both proteins have visible spectra similar to that of the low-spin sulfite reductase of Desulfovibrio vulgaris strain Hildenborough and no band at 715 nm, characteristic of high-spin Fe3+ complexes in isobacteriochlorins is observed. EPR shows that as isolated the siroheme is in a low-spin ferric state (S = 1/2) with g-values at 2.40, 2.30 and 1.88 for the Methanosarcina barkeri enzyme and g-values at 2.44, 2.33 and 1.81 for the Desulfuromonas acetoxidans enzyme. Chemical analysis shows that both proteins contain one siroheme and one [Fe4S4] center per polypeptidic chain. These results suggest that the low molecular weight, low-spin non-heme iron siroheme proteins represent a new homologous class of sulfite reductases common to anaerobic microorganisms. PMID- 3028381 TI - Role of pertussis toxin sensitive G proteins in the alpha 1 adrenergic receptor but not in the thyrotropin receptor mediated activation of membrane phospholipases and iodide fluxes in FRTL-5 thyroid cells. AB - The regulation of thyroid hormone formation by thyrotropin and norepinephrine involves the activation of both phospholipases C and A2. When FRTL-5 cells are incubated with 10(-10)M pertussis toxin for 4 to 20 h, the stimulation of iodide efflux by norepinephrine is inhibited by 50 to 70%. At the same toxin concentration the norepinephrine induced increase in cytosolic Ca2+ is unaffected; however upon 20 h pretreatment with 10(-9)M pertussis toxin a 30% inhibition is observed. By contrast, the pertussis toxin treatment had no effect on the increase in iodide efflux or in cytosolic Ca2+ levels induced by thyrotropin. Our data suggest that two GTP binding proteins sensitive to pertussis toxin are involved in the alpha 1 adrenergic but not in the thyrotropin induced activation of the signal transduction mechanisms leading to iodide efflux in FRTL-5 cells. PMID- 3028383 TI - Multiple molecular forms of interferon display different specific activities in the induction of the antiviral state and 2'5' oligoadenylate synthetase. AB - Both Hu IFN-alpha A and Hu IFN-alpha D, produced by two independent recombinant bacterial clones, are mixtures of monomers, dimers and trimers. These forms, when assayed individually in heterologous MDBK cells, induced different degree of antiviral and 2'5' oligoadenylate synthetase (2'5' A synthetase) activities: the antiviral activity of the monomer is greater than that of the dimer and the trimer, whereas the activity of 2'5' A synthetase induction is lower with the monomer than with the dimer or the trimer. Similar differences are also observed on human cells. Compared to the mononeric form, the dimeric and the trimeric forms of Hu IFN-alpha A show higher antiviral inducing activity on heterologous MDBK cells than on homologous WISH cells, whereas the 2'5' A synthetase inducing activity in these two cell lines is about the same. Thus for the same antiviral activity, the trimer or the dimer compared to the monomer are much better inducers of the 2'5' A synthetase on human than on MDBK cells. PMID- 3028384 TI - Enhancement of cmyc mRNA concentration in dog thyrocytes initiating DNA synthesis in response to thyrotropin, forskolin, epidermal growth factor and phorbol myristate ester. PMID- 3028385 TI - Identification of calmodulin activity in purified retroviruses. AB - Several viruses have been shown to require calcium for their function, and to bind calcium at specific sites. However, the nature of the calcium binding molecule on viruses has not been established. One possibility is the ubiquitous calcium-binding protein calmodulin. Our studies were designed to determine whether feline leukemia virus contained calmodulin. Accordingly, we tested purified feline leukemia virus for the presence of calmodulin-like activity. The virus, like authentic calmodulin, activated cyclic AMP phosphodiesterase. The ability of the virus to activate the enzyme was blocked in the presence of the known calmodulin inhibitors trifluoperazine and W-7. This indirect evidence for the presence of calmodulin was confirmed by radioimmunoassay. Several other retroviruses were also tested using radioimmunoassay and found to contain calmodulin. Our results indicate that the calcium binding site in retroviruses may be calmodulin. PMID- 3028386 TI - Hydroperoxide-induced radical production in liver mitochondria. AB - When isolated rat liver mitochondria are treated with tert-butyl hydroperoxide in the presence of the spin trap 5,5-dimethyl-1-pyrroline-N-oxide, a six-line ESR signal is observed with parameters characteristic of a carbon-centered radical. The radical is shown to be CH3. using 2-methyl-2-nitrosopropane as the spin trap. Inhibition of radical production by EDTA and N-ethylmaleimide provides evidence for participation by metals and reduced sulfhydryl groups in the radical generating reaction. It is proposed that radicals are formed through the reaction between a reducing agent, a metal and the hydroperoxide. PMID- 3028387 TI - Localization of enzyme for heme attachment to apocytochrome c in yeast mitochondria. AB - Fractionation of yeast mitochondria by controlled hypotonic treatment revealed that the enzyme for heme attachment to apocytochrome c was localized in mitochondrial inner membrane. Trypsin digestion of mitoplasts resulted in a considerable loss of enzymatic activity, whereas the enzyme in intact mitochondria resisted the digestion. Triton X-100 solubilized the enzyme from the membrane but high concentration of salt did not. These results reveal that the enzyme for heme attachment is localized in mitochondrial inner membrane facing the cytoplasmic surface. PMID- 3028388 TI - Maintenance of autonomous genetic elements in twelve transgenic mouse strains established after transfer of pPyLT1 DNA. AB - Maintenance and efficient meiotic segregation of autonomous genetic elements in four transgenic mouse strains established after micro-injection of plasmid pPyLT1 were previously reported. These findings are now extended to a total of twelve independent transgenic families. In spite of extensive rearrangements, half of these plasmids maintained the region of pBR322 necessary for shuttle transfer in E. coli. They all included mouse DNA sequences and the same, or closely related sequences were present in distinct mouse strains. PMID- 3028389 TI - Beta-1,3-glucan receptor and peptidoglycan receptor are present as separate entities within insect prophenoloxidase activating system. AB - Silkworm plasma was passed over a peptidoglycan-Sepharose 4B column or a CPB column [CPB, curdlan type polysaccharide (beta-1,3-glucan) bead] in the absence of divalent cation and the effluents from the columns were named plasma-PG and plasma-CPB, respectively. Prophenoloxidase activating system in plasma-PG was triggered by beta-1,3-glucan but not by peptidoglycan and the system in plasma CPB was triggered by peptidoglycan but not by beta-1,3-glucan, suggesting that the peptidoglycan-Sepharose 4B column and the CPB column remove peptidoglycan receptor and beta-1,3-glucan-receptor, respectively, from plasma. This result indicates that both receptors exist as separate entities in silkworm plasma. It is suggested that plasma-PG and plasma-CPB may be used as specific reagents to detect minute amounts of beta-1,3-glucan and peptidoglycan. PMID- 3028390 TI - Differences in phospholipid incorporation of 32P relevant to alpha 1-receptor coupling events in rat and rabbit aorta. AB - Labelling of membrane phospholipids with 32P was compared in rat and rabbit aorta under basal conditions and during alpha 1-receptor stimulation. Incorporation of 32P proceeded at a significantly higher rate in rat tissue. The ratio of basal labelling following 30 min of incubation for rat/rabbit arteries was 4.8 for phosphatidylinositol diphosphate (PIP2), 6.0 for phosphatidylinositol phosphate (PIP), 9.0 for phosphatidylinositol (PI), 6.0 for phosphatidic acid (PA) and 18.7 for phosphatidylcholine (PC). Addition of 10(-5)M norepinephrine (NE) to labelled tissues resulted in a similar decrease in [32P]-PIP2 in both rat and rabbit tissues. Greater percent increases were seen in rabbit tissue of [32P]-PA (4-6 fold), and [32P]-PI (3-5 fold), when measured over the initial 10 minutes of agonist exposure. While NE caused a gradual increase of 32P incorporation into PC in rabbit aorta, reaching 180% above control after 10 minutes, PC labelling was not increased in rat aorta. Our findings provide evidence for the enhanced labelling of rat vs rabbit aorta phospholipids. This may account for differences in receptor responses and associated Ca+ movements which have been previously recognized to exist between aorta of these two species. PMID- 3028391 TI - Acute effects of alpha-MSH on the rat zona glomerulosa in vivo. AB - alpha-MSH acutely enhanced the plasma concentration of aldosterone (but not that of corticosterone) in the rat, with a maximal response at a dose of 100 micrograms/kg. This dose of alpha-MSH increased the blood level o aldosterone and the activity of 11 beta-hydroxylase and 18-hydroxylase of capsular adrenals in rats infused for 24 h with dexamethasone, dexamethasone plus ACTH, or captopril plus angiotensin II, but not in animals treated with captopril alone. The plasma concentration of corticosterone and the activity of 11 beta-hydroxylase in the inner adrenal layers were not changed. These findings indicate that alpha-MSH is specifically involved in the acute stimulation of the late steps of the secretory activity of the rat zona glomerulosa, and that this action of alpha-MSH requires a normal level of circulating angiotensin II. PMID- 3028392 TI - Expression of IL-1 genes in human and bovine chondrocytes: a mechanism for autocrine control of cartilage matrix degradation. AB - In this report we describe the presence of interleukin-1 activity in medium conditioned by bovine articular cartilage. Preparations partially purified by Sephacryl S200 chromatography (Mr 18000-25000) stimulate murine thymocyte proliferation in the lymphocyte activation factor assay. Furthermore, the factor(s) activate cartilage tissue to secrete a protease which is essential for the activity of purified synovial collagenase. We also demonstrate the presence of mRNA coding for IL-1 alpha and beta in human articular chondrocytes and conclude that the human monocytic and chondrocytic mRNAs are identical. Our results demonstrating cartilage expression of IL-1 genes suggest the possibility of an autocrine mechanism whereby chondrocyte production of matrix degrading proteases is initiated by chondrocyte derived IL-1. PMID- 3028393 TI - Anomer specificity of glucose-6-phosphatase and glucokinase. AB - The anomeric form of glucose produced by glucose-6-phosphatase was studied using an apparatus that specifically measures beta-D-glucose. The time course of beta-D glucose formation from glucose-6-P by glucose-6-phosphatase is essentially linear. In the presence of mutarotase, this rate is reduced to 70% of that obtained in the absence of mutarotase. When detergent treated microsomes were used, the rate of beta-D-glucose formation is unaffected by mutarotase. These results suggest that only beta-anomer of glucose is produced by microsomal glucose-6-phosphatase and this specificity is determined by translocase for glucose-6-P or glucose. It was also demonstrated that alpha-D-glucose is the substrate for glucokinase. PMID- 3028394 TI - Synthesis in yeast of hepatitis B virus surface antigen modified P31 particles by gene modification. AB - The pre-S2 portion of hepatitis B virus surface antigen P31 gene was modified to make gene products resistant to trypsin-like proteases in Saccharomyces cerevisiae. The coding sequence for 6 amino acids (Ser44 - Thr49) including Arg48 was removed, and the altered gene was inserted into an expression vector. The modified HBsAg P31 (M-P31c) gene products, consisting of GP37 and GP34, formed particles having both HBsAg antigenicity and polymerized-albumin receptor activity. Since the M-P31c particles can elicite two kinds of protective antibodies against hepatitis B virus, anti-S and anti-pre-S2 antibodies, the M P31c particles are expected to be potentially effective to S-nonresponders. PMID- 3028395 TI - Bovine angiotensin-converting enzyme: amino-terminal sequence analysis and preliminary characterization of a hybridization-selected primary translation product. AB - Bovine lung angiotensin-converting enzyme was isolated in pure form and the sequence of the first twenty-two NH2-terminal amino acids determined. Oligonucleotides, complementary to a selected portion of the NH2-terminal amino acid sequence of the bovine glycoprotein (Mr 145,000), were synthesized and used for hybridization selection of angiotensin-converting enzyme mRNA. The hybridization-selected mRNA programmed the in vitro synthesis of a single polypeptide (Mr 130,000) that was specifically immunoadsorbed by anti-bovine enzyme antibodies. Preliminary sequence analysis of the primary translation product suggests that bovine angiotensin-converting enzyme is synthesized without a transient NH2-terminal signal sequence. PMID- 3028396 TI - Regulation of the nah and sal operons of plasmid NAH7: evidence for a new function in nahR. AB - The catabolism of naphthalene and salicylate is specified by two operons on an 80 Kb metabolic plasmid, NAH7. These operons, nah and sal, are carried on the contiguous 30 Kb EcoRI-A, C fragments, and are under positive control of a regulator region, nahR. Five Nah Sal Tn5 insertion mutants form two complementation groups: A = nahR203, nahR204; and B = nahR201, nahR202, nahR205. The physical and genetic maps assign the nahR location to the 15.7-17.2 Kb region of the EcoRI-A fragment, with suggestion of more than one control gene. PMID- 3028397 TI - Dibutyryl-cyclic AMP inhibits cholesterol esterification in J 774 monocyte-like cells. AB - The effect of dibutyryl-cyclic AMP (dbcAMP) and theophylline was investigated on oleic acid incorporation into cholesteryl esters and triacylglycerols in the mouse monocyte-macrophage cell line J 774. 24h pretreatment of macrophages with dbcAMP decreased cholesteryl ester formation in a dose-dependent manner (about 4 fold reduction for dbcAMP 10(-4)M + theophylline 10(-3)M), while oleic acid incorporation into triacylglycerols was markedly (2 to 3 fold) enhanced. The catabolism of acetylated LDL was only slightly affected (about 15-20% reduction with dbcAMP 5 X 10(-4)M + theophylline 10(-3)M). Acyl Coenzyme A: cholesterol-O acyl-transferase activity, measured in vitro on cell homogenates, was reduced in dbcAMP-treated cells, whereas diacylglycerol acyltransferase activity was increased. These results suggest that cyclic AMP can modulate cholesteryl ester and triacylglycerol formation in macrophages, and that these metabolisms are inversely regulated. Agents which increase cyclic AMP intracellular level could be of interest for reducing cholesteryl ester accumulation in macrophages. PMID- 3028399 TI - Angiotensin-induced formation and metabolism of inositol polyphosphates in bovine adrenal glomerulosa cells. AB - The actions of angiotensin II (AII) on inositol polyphosphate production and metabolism were analyzed in cultured bovine adrenal glomerulosa cells. In cells labeled for 24 hr with [3H]inositol, AII caused a rapid and prominent rise in formation of Ins-P3 (mainly the Ins-1,3,4,-P3 isomer) and of Ins-P4, with marked increases in two isomers of Ins-P2 and Ins-P. These findings are consistent with rapid formation and turnover of Ins-1,4,5-P3, partly via conversion to Ins 1,3,4,5-P4 with subsequent metabolism to Ins-1,3,4-P3 and lower inositol phosphates. The demonstration of a cytosolic Ins-P3-kinase gave further evidence for the presence of the tris/tetrakisphosphate pathway and Ins-P4 synthesis during AII action in the bovine adrenal cortex. PMID- 3028400 TI - An ATP/2e-stoichiometry of 1 1/2 is thermodynamically possible for site 3 of oxidative phosphorylation. AB - Free energy changes for ATP synthesis (delta GP) and 2e(-)-transfer across Site 3 (delta GR) were determined during oxidative phosphorylation by rat liver mitochondria. At static head, -delta GR/delta GP ranged narrowly between 1.55 and 1.59 with five different respiratory substrates. Thus, an ATP/2e- of 1 1/2 at Site 3 is thermodynamically possible with regards to overall reactants and products. Using nonequilibrium thermodynamics, phenomenological stoichiometries were close to 1 1/2 for all substrates suggesting that ATP/2e- at Site 3 is, in fact, 1 1/2. An ATP/2e- of 1 1/2 can only be possible if H+/O is 4 for cytochrome oxidase. PMID- 3028398 TI - Both 2',3'-dideoxythymidine and its 2',3'-unsaturated derivative (2',3' dideoxythymidinene) are potent and selective inhibitors of human immunodeficiency virus replication in vitro. AB - 2',3'-Dideoxythymidine (ddThd) and its 2',3'-unsaturated derivative 2',3' dideoxythymidinene (ddeThd) are potent and selective inhibitors of human immunodeficiency virus (HIV) in vitro. When evaluated for their inhibitory effects on the cytopathogenicity of HIV in MT-4 cells, ddThd and ddeThd completely protected the cells against destruction by the virus at a concentration of 1 microM and 0.04 microM, respectively. In this aspect, ddeThd was about 5 times more potent than 2',3'-dideoxycytidine (ddCyd), one of the most potent and selective anti-HIV compounds now pursued for its therapeutic potential in the treatment of AIDS. ddThd and ddeThd also suppressed HIV antigen expression at 1 microM and 0.04 microM, respectively. Their selectivity indexes, as based on the ratio of the 50% cytotoxic dose to the 50% antiviral effective dose, were 120 (ddeThd) and greater than 625 (ddThd). PMID- 3028401 TI - A reassessment of the product specificity of the NADPH:O2 oxidoreductase of human neutrophils. AB - Native ferricytochrome c, but not acetylated ferricytochrome c, stimulates the flow of electron equivalents passing through the neutrophil NADPH:O2 oxidoreductase complex. At 28 mM it increases NADPH oxidase activity by 157 +/- 15% (n = 5) over that measured in its absence. Enhanced activity is predominantly seen in oxidoreductase-rich 27,000 X g membrane preparations obtained from phorbol myristate acetate activated cells. Superoxide formation is also enhanced. Although some of the stimulatory activity seen with addition of native ferricytochrome c to oxidoreductase-rich membrane suspensions might have been explained in terms of mitochondrial contamination, this was ruled out. Comparable membrane preparations from resting cells were devoid of NADPH oxidase activity. Azide, a well-known inhibitor of the electron transport chain, did not block the enhancing effect of native ferricytochrome c. These results indicate that native ferricytochrome c is not a suitable scavenger of superoxide in quantitating the product specificity of the oxidoreductase since it amplifies the apparent rate of superoxide formation with respect to measured rates of NADPH oxidation conducted in its absence. By using acetylated ferricytochrome c in place of native ferricytochrome c in quantitating the product specificity of the oxidoreductase we show that no more than 70% of the electron equivalents donated by NADPH to the oxidoreductase are involved in superoxide formation. The remaining 30% of the electron equivalents given up by NADPH to the oxidoreductase appear to be involved in direct formation of hydrogen peroxide. PMID- 3028403 TI - Recombinant tumor necrosis factor and interleukin-1 both stimulate human synovial cell arachidonic acid release and phospholipid metabolism. AB - Stimulation of [3H]-arachidonic acid labeled human synovial cells with 3.0 X 10( 10)M recombinant interleukin-1 or tumor necrosis factor resulted in the release of incorporated radiolabel (41.1% and 27.7% respectively). Analysis of [3H] arachidonic acid labeled phospholipids showed that interleukin-1 and tumor necrosis factor diminished [3H]-arachidonic acid in phosphatidyl-choline phosphatidylserine and phosphatidylinositol. Treatment of [3H]-arachidonic acid labeled cells with 10(-3)M dibutyryl cyclic AMP or 10(-6)M PGE2 did not affect spontaneous or stimulated [3H] arachidonic acid release significantly. These data show that recombinant interleukin-1 and tumor necrosis factor stimulate human synovial cell phospholipase activity in a similar manner and that this activity is not affected significantly by agents that elevate cyclic AMP. PMID- 3028402 TI - The cardiac gap junction protein (Mr 47,000) has a tissue-specific cytoplasmic domain of Mr 17,000 at its carboxy-terminus. AB - The molecular weight of the heart gap junctional protein subunit was, until recently, believed to be about Mr 28,000-30,000, similar to that of other previously characterized gap junctional proteins. A larger polypeptide of about Mr 44,000-47,000, which undergoes proteolysis during isolation, has recently been proposed as the form of the heart junction protein in vivo. We show here that this entity has the same amino-terminal sequence as the previously characterized Mr 29,000-30,000 component. Thus, the cardiac junctional protein has, at its carboxy-terminus, cytoplasmic domain of Mr 17,000; this domain is absent in the liver protein. These observations provide further evidence that gap junction proteins form a highly diversified family. PMID- 3028404 TI - Regulation of membrane associated protein kinase C activity by guanine nucleotide in rabbit peritoneal neutrophils. AB - Evidence is shown that protein kinase C is the major kinase which can phosphorylate histone H-1 in a membrane fraction prepared from rabbit peritoneal neutrophils. Addition of phorbol-12-myristate-13-acetate (PMA) (0.1 microgram/ml) or guanosine-5'-(3-O-thio)triphosphate (GTP gamma S) (10 microM) to the membrane fraction results in an increase of the phosphorylation of histone H-1. To achieve this effect, calcium (20 microM) is required for GTP gamma S but not for PMA. The effect of GTP gamma S, but not PMA is inhibited in membranes obtained from cells pretreated with pertussis toxin. The kinase activity is also enhanced by treatment of the membrane with 10 microM of GppNHp or GTP but not with GDP, GMP, cGMP, ATP, ADP, AMP and cAMP. This is the first direct evidence that a GTP binding protein is involved in the activation of membrane associated protein kinase C. PMID- 3028405 TI - The type-1 protein phosphatase activating factor FA is a membrane-associated protein kinase in brain, liver, heart and muscles. AB - Although the activating factor FA of the type-1 protein phosphatase has long been recognized as a cytosolic enzyme involved in the regulation of cell metabolism and nervous functions, strong indications have been obtained that FA is in fact a membrane-bound protein kinase in most mammalian tissues. For instance, direct treatment of the tissue extracts including brain, liver, cardiac, smooth and skeletal muscles with 1% Triton X-100 can cause several fold stimulation of the FA activity. Moreover, at least 50% of the FA can be detected in the particulate fractions of the extracts. Chromatography of the extracts in the presence and absence of Triton X-100 further demonstrate these results. The data can now explain the reason why most people can not isolate reasonable amount of FA from mammalian tissues. It is recommended that Triton X-100 should be used for purification of FA from most mammalian tissue extracts. The results also suggest that most previous studies on the action of FA involved in the regulation of cell functions should be re-evaluated and the membrane-associated FA should be taken into consideration. PMID- 3028406 TI - Comparison of the roles of calmodulin and protein kinase C in activation of the human neutrophil respiratory burst. AB - The roles of calmodulin and protein kinase C in the activation of the human neutrophil respiratory burst were characterized pharmacologically. The protein kinase C inhibitors 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine (H-7) and N-(2 aminoethyl)-5-isoquinolinesulfonamide (H-9) did not inhibit superoxide anion generation by neutrophils stimulated for 30 minutes with N-formyl-L-methionyl-L leucyl-L-phenylalanine (FMLP) or 4 beta-phorbol 12 beta-myristate 13 alpha acetate (PMA). However, H-7 did depress superoxide production during the first 5 minutes following stimulation. In contrast, the specific calmodulin antagonist N (6-aminohexyl)-5-chloro-1-naphthalenesulfonamide (W-7) and the dual calmodulin antagonist/protein kinase C inhibitor trifluoperazine (TFP) were potent inhibitors of the response throughout the 30 minute incubation. Stimulation of neutrophils with submaximal doses of FMLP or PMA failed to promote inhibition of the respiratory burst by H-7 or H-9, but did stimulate a respiratory burst response which was not inhibited by TFP or W-7. These results suggest that while protein kinase C may play a role in the initiation of the respiratory burst response, propagation of the response is dependent on calmodulin-dependent processes. The inability of TFP and W-7 to inhibit superoxide anion generation in response to submaximal stimulatory doses of FMLP or PMA suggests that calmodulin independent processes may also be involved in activation of the respiratory burst. PMID- 3028407 TI - Cholesterol efflux from cultured adipose cells is mediated by LpAI particles but not by LpAI:AII particles. AB - Cholesterol efflux was studied in cultured adipose cells after preloading with LDL cholesterol. Long-term exposure to LpAI and LpAI:AII particles isolated from the HDL fraction showed that LpAI particles only were able to promote cholesterol efflux. Liposomes containing different ApoAI/ApoAII molar ratios were tested: the larger the proportion of ApoAI, the faster the ability to remove cholesterol from Ob1771 cells. Dose-response curves showed that LpAI particles were active within a physiological range of concentrations, whereas LpAI:AII particles had no effect at all concentrations. The results are in favour of LpAI particles being the active components of the HDL fraction for the promotion of cholesterol efflux and suggest that LpAI particles and LpAI:AII particles represent distinct metabolic entities. PMID- 3028408 TI - Cloning and regulation of rat apolipoprotein B mRNA. AB - Recombinant cDNA clones that code for apolipoprotein B(apoB) were isolated from a rat liver cDNA library, using synthetic oligonucleotide probe derived from the sequence of human apoB cDNA. The nucleotide and deduced amino acid sequences of the rat apoB clone pRB5, 1.2 kb in length, showed 83% and 84% homology to those of human apoB. Northern blot analysis revealed that rat apoB cDNA probe cross reacts with human and rabbit apoB mRNA sequences and the size of those mRNAs, approximately 15 kb long, were not discernibly different. In addition, apoB mRNA was abundant only in the liver and intestine. Finally, cholesterol feeding to rats for six weeks resulted in a several-fold increase in the level of apoB mRNA in the liver. PMID- 3028409 TI - NAD-dependent hydrogenase from Alcaligenes eutrophus Z1: does it have a regulatory centre? AB - Evidence is presented for the existence of a relatively high-potential regulatory centre in the NAD-dependent hydrogenase from the hydrogen oxidizing bacterium Alcaligenes eutrophus Z1. Reduction of the hydrogenase to the redox potentials lower than -100 mV converts the enzyme into a catalytically active state that is remarkably stable to oxidants. Once activated, the enzyme does not loose its activity on intensive oxygenation for at least 3 hours. A novel hydrogenase ESR signal with a wide temperature optimum and a approximately -100 mV midpoint redox potential was detected. We suggest that the reduction of this redox centre trigger conformational changes in the inactive oxidized enzyme molecule, thus reorganizing the latter into the active one. PMID- 3028410 TI - Leukotrienes and related eicosanoids are produced by frog leukocytes. AB - The metabolites of inflammatory cells produced by massive infection of the peritoneal cavity of two related European species of frogs, Rana temporaria and Rana arvalis were examined for lipoxygenase-generated products of exogenous arachidonic acid. Cells of Rana temporaria produced large amounts of 5 hydroxyeicosatetraenoic acid and leukotriene B4. Cells from Rana arvalis produced only 15-hydroxyeicosatetraenoic acid. This is the first unequivocal demonstration of such enzyme activity in lower vertebrates. There was a trend towards increased mortality in the species without evidence of 5-lipoxygenase activity. PMID- 3028411 TI - Mitochondrial function after acute alteration of the endogenous insulin-to glucagon ratio. AB - Mannoheptulose (2g/kg i.p.) increases serum glucagon and decreases serum insulin via its effect on pancreatic islet cells. These changes in endogenous hormone status had effects on rat liver mitochondria that were comparable to the effects of injecting porcine glucagon (0.5 mg/kg i.p.). Mitochondrial adenine nucleotide content was increased 38 or 39% by mannoheptulose or glucagon respectively, citrulline synthesis by 165 or 193%, pyruvate carboxylation by 113 or 135%, coupled respiration by 34 or 42%, and uncoupled respiration by 40 or 54%. We conclude that the reciprocal changes in endogenous insulin and glucagon brought about by mannoheptulose offer a useful and interesting alternative to glucagon injection for studying the effects of these pancreatic hormones on liver mitochondria. PMID- 3028412 TI - Phosphorylation of the alpha 1,2-linked mannosylsaccharide by N-acetylglucosamine 1-phosphotransferase from fibroblasts occurs at the terminal mannose. AB - UDP-GlcNAc: Lysosomal enzyme precursor N-acetylglucosamine-1-phosphotransferase activity from normal fibroblasts was measured using methyl 2-O-(alpha-D mannopyranosyl) 6-deoxy-6-fluoro-alpha-D-mannopyranoside and methyl 2-O-(6-deoxy 6-fluoro-alpha-D-mannopyranosyl) alpha-D-mannopyranoside as acceptors. The results indicate that the phosphorylation in man alpha 1----2 man sequence occurs at the C-6 position of the terminal mannose residue. PMID- 3028413 TI - Spin-trapping of sulfite radical anion, SO3-., by a water-soluble, nitroso aromatic spin-trap. AB - Sulfite radical anion, SO3-., which is generated either by non-enzymatic reaction of hydrogen peroxide (H2O2-) with sulfite (SO3(2-)) or by the oxidation of bisulfite (HSO3) with Ce4+ ion, can be trapped with a water-soluble, nitroso aromatic spin-trap, sodium 3,5-dibromo-4-nitrosobenzenesulfonate (DBNBS, 1), yielding an ESR spectrum with coupling constants [aN (1) = 12.9 G, aH (2) = 0.8 G] and a g-value of 2.0063. The SO3- radical adduct (spin adduct) was observed even in the presence of the very low concentration of H2O2 (1.21 X 10(-2) mumol). PMID- 3028414 TI - K-252 compounds, novel and potent inhibitors of protein kinase C and cyclic nucleotide-dependent protein kinases. AB - K-252 compounds (K-252a and b isolated from Nocardiopsis sp. (1) and their synthetic derivatives) were found to inhibit cyclic nucleotide-dependent protein kinases and protein kinase C to various extents. The inhibitions were of the competitive type with respect to ATP. K-252a was a non-selective inhibitor for these three protein kinases with Ki values 18-25 nM. K-252b showed a comparable potency for protein kinase C (Ki, 20nM), whereas inhibitory potencies for cyclic nucleotide-dependent protein kinases were reduced. KT5720 and KT5822 selectively inhibited cAMP-dependent (Ki, 60nM) and cGMP-dependent (Ki, 2.4nM) protein kinases, respectively. PMID- 3028415 TI - Increased adenylate cyclase activity in rat thyroid epithelial cells expressing viral ras genes. AB - The activity of the adenylate cyclase catalytic subunit is higher in Harvey and Kirsten Murine Sarcoma Viruses-infected thyroid epithelial cells than in uninfected control cells either in the presence of Mg2+ alone or following stimulation by Mn2+ or forskolin. The higher activity is associated with an increased cAMP cellular content. The Gpp(NH)p and F- anion are more effective positive modulators in the control than in the virus infected cells: these results exclude therefore that the ras p21 proteins can act as the G-protein alpha-subunit and suggest that they negatively interfere with the G-protein modulation of the adenylate cyclase system. PMID- 3028416 TI - In-vitro modulation of protein kinase C activity by environmental chemical pollutants. AB - A number of environmental chemical pollutants have been reported to cause tumors or help in the propagation of tumors in experimental animals. The in-vitro effects of a few chemical contaminants were studied on the histone phosphorylation and 3H Phorbol dibutyrate (PdBu) binding of partially purified Ca2+/phospholipid dependent protein kinase c (PKC) from the brains of Fischer F344 and B6C3F1 mice. The enzyme was prepared by a modified method which gave approximately 75-fold purification. A differential effect of various compounds was observed on the phosphorylation activity and PdBu binding of PKC from rats and mice. The reported tumor promoting ability and effect on protein kinase C activity appeared to be related in the case of the rat enzyme, although causality cannot be inferred. PMID- 3028417 TI - Evidence for a role of a vicinal dithiol in catalysis and proton pumping in mitochondrial nicotinamide nucleotide transhydrogenase. AB - The effect of glutathione, glutathione disulfide and the dithiol reagent phenylarsine oxide on purified soluble as well as reconstituted mitochondrial nicotinamide nucleotide transhydrogenase from beef heart was investigated. Glutathione disulfide and phenylarsine oxide caused an inhibition of transhydrogenase, the extent of which was dependent on the presence of either of the transhydrogenase substrates. In the absence of NADPH glutathione protected partially against inactivation by glutathione disulfide and phenylarsine oxide. In the presence of NADPH glutathione also inhibited transhydrogenase. Reconstituted transhydrogenase vesicles behaved differently as compared to the soluble transhydrogenase and was partially uncoupled by GSSG. It is concluded that transhydrogenase contains a dithiol that is essential for catalysis as well as for proton translocation. PMID- 3028418 TI - Membrane-bound phosphotyrosyl-protein phosphatase activity in human erythrocytes. Dephosphorylation of membrane band 3 protein. AB - Human erythrocyte membranes exhibit, in addition to "acid" p-nitrophenyl phosphatase activity, remarkable phosphotyrosyl-protein phosphatase activity, assayed on synthetic polymer poly (Glu-Tyr) 4:1, previously phosphorylated on Tyr residues by rat spleen tyrosine-protein kinase. The results reported here indicate that such a 32P-Tyr-phosphatase activity, rather than p-nitrophenyl phosphatase, is involved in the dephosphorylation of transmembrane band 3 protein on 32P-tyrosine residues. PMID- 3028420 TI - Endonucleolytic activities of nuclei from rat ascites hepatoma. AB - By autodigestion (endogenous endonucleolysis) of rat liver (RL) or rat-ascites hepatoma (AH) nuclei, the nucleosomes were released from the RL, but not from the AH, nuclei. In contrast, by micrococcal nuclease digestion (exogenous endonucleolysis), the nucleosomes were released more rapidly from the AH than from the RL nuclei. A 0.6 M NaCl extract of the RL or AH nuclei was filtered through a Sephadex G-100 column. The resulting topoisomerase fraction was subjected to DNA relaxation and catenation assays with pBR322 DNA as a substrate. Consequently, the relaxation activity was almost the same between the RL and AH fractions, whereas the catenation activity was much higher in the AH fraction. PMID- 3028419 TI - A novel proenkephalin processing carboxypeptidase and its activation by cyclic AMP dependent protein kinase. AB - Carboxypeptidase B-like enzymes cleaving Met-enkephalin-Arg from synaptosomes of the rat striatum purified using a DEAE-cellulose column and Met-Arg-CH-Sepharose 4B affinity column proved to be different from enkephalin-convertase, lysosomal carboxypeptidase B-like enzyme, pancreas carboxypeptidase B and carboxypeptidase Y, in effects of inhibitors and activators, pH optimum (7.5-8.5) and molecular size (50,000). This enzyme, named "Processin CP-E" was activated by cAMP dependent protein kinase, and the Vmax was increased from 4.3 to 13.3 microM/min/mg protein, while the Km (28.2 microM) was unchanged. PMID- 3028421 TI - Effects of sodium butyrate and dibutyryl cyclic AMP on thermosensitivity of HeLa cells and their production of heat shock proteins. AB - When HeLa cells were incubated at 42 degrees C for 6 h with 1 mM sodium butyrate or when cells treated with 1 mM dibutyryl cyclic AMP for 24 h were incubated at 42 degrees C for 6 h, they were more thermoresistant than heated control cells without such drugs. The production of heat shock proteins was not enhanced by the drug treatment. These results suggest that there is a factor (or factors) other than heat shock proteins that accounts for the thermoresistance of HeLa cells. PMID- 3028422 TI - Characterization of intra- and extracellular transforming growth factors from normal and avian sarcoma virus-transformed rat cells. AB - Intra- and extracellular transforming growth factors (TGFs) have been characterized in an avian sarcoma virus-transformed rat cell line, 77N1, and the nontransformed parental cell line, NRK. Serum-free conditioned medium from 77N1 and cell extracts from NRK and 77N1 were subjected to ion exchange column chromatography. Intracellular TGFs in cell extracts of NRK and 77N1 cells showed a major peak of DNA synthesis-stimulating and colony-forming activities which eluted in the 0.10 to 0.15 M salt concentration region. Extracellular TGF in conditioned medium of 77N1 cells showed a major peak of activity which eluted at 0.05 to 0.06 M salt concentration. Furthermore partially purified extracellular TGF had a molecular weight of about 40,000 daltons, whereas that for intracellular TGF was about 12,000 daltons. These intra- and extracellular TGFs, as well as TGF gamma 2 which was purified from 77N1 cell extract, were heat- and acid-labile polypeptides sensitive to treatment with dithiothreitol. Radioimmunoprecipitation analyses with antiserum against TGF gamma 2 demonstrated that intra- and extracellular TGFs in 77N1 and NRK cells were immunologically identical or very closely related to each other and suggested the possibility that extracellular TGF from 77N1 cells was released into serum-free conditioned medium after formation of complex with other cellular components. PMID- 3028423 TI - Activation of oncogenes by transposable elements. AB - Mammalian DNA contains several families of highly repeated sequences, some of which have been suggested to be mobile elements. We have screened tumour tissue for the rearrangement of cellular oncogenes and found evidence for the behaviour of repetitive DNA sequences as transposable elements which may activate oncogenes. In the mouse myeloma NSI and XRPC24 we found that intracisternal A particle genome was inserted into the coding region of c-mos. In both cases the rearranged c-mos was transcriptionally activated and was also able to transform NIH 3T3 cells. In the canine transmissible venereal tumour we found that c-myc was rearranged due to the insertion of an 1.8 kilobase pair cellular DNA. Nucleotide sequence analysis demonstrated that the inserted piece is 60% homologous to the monkey KpnI element which is a representative of the LINE group. PMID- 3028425 TI - Sodium stibogluconate (Pentostam) inhibition of glucose catabolism via the glycolytic pathway, and fatty acid beta-oxidation in Leishmania mexicana amastigotes. AB - The biochemical mechanism of action of antimony (Sb) in pentavalent form complexed to gluconic acid (sodium stibogluconate)--the drug of choice for the leishmaniases--has been only slightly investigated. We recently reported that, in stibogluconate-exposed Leishmania mexicana amastigotes, there is a dose-dependent decrease in the ATP/ADP ratio [Berman et al., Antimicrob. Agents Chemother. 27, 916 (1985)]. To investigate mechanisms by which ADP phosphorylation to ATP might be inhibited, stibogluconate-exposed amastigotes were incubated with [14C]glucose, fatty acid, or acetate, and 14CO2 production was determined. In organisms exposed to 500 micrograms Sb/ml, formation of 14CO2 from [6-14C]glucose and [1-14C]palmitate was inhibited 69 and 67% respectively. In comparison, formation of 14CO2 from [1-14C]glucose and [2-14C]acetate was inhibited less than 15%. These results suggest that glucose catabolism via glycolytic enzymes and fatty acid beta-oxidation, but not glucose metabolism via the hexosemonophosphate shunt or the citric acid cycle, is specifically inhibited in stibogluconate exposed Leishmania mexicana amastigotes. Inhibition of these pathways suggests a mechanism for the inhibition of ADP phosphorylation previously reported. PMID- 3028424 TI - Hormone receptor regulated proopiomelanocortin gene expression. AB - The activity of the POMC gene is regulated by both stimulatory and inhibitory hormones. Presumably, some balance exists in the influence of these hormones in order to maintain a steady-state level of activity of this gene. Physiological insults, such as stress, may upset this balance and change the rate of production of POMC and its biologically active peptides. The relative strength of these different hormones may therefore determine the long-term expression of this gene. Chronic administration of CRF to rats, primates and humans produces prolonged increases in plasma ACTH levels. This long-term effect is most likely due to an activation of the POMC gene. Interestingly, chronic treatment of anterior pituitary cells with CRF desensitizes CRF receptors. Thus, despite the corticotrophs becoming refractory to the acute stimulatory actions of CRF, the POMC gene remains stimulated. These findings suggest that corticotrophs do not have to be continuously activated by CRF to maintain a long-term increase in POMC gene expression. This contrasts with the actions of glucocorticoids whose effects are abruptly terminated following their removal from the target tissue. The molecular basis of this form of cellular memory to the actions of CRF may involve cAMP regulatory phosphoproteins binding to and activating the POMC gene. If this phenomenon is shown to occur and the phosphorylation state of these nuclear proteins is found to govern their level of interaction with POMC gene, then it would represent a novel mechanism of gene regulation. Proof for this mechanism and the elucidation of how other second messengers such as protein kinase C and calcium regulate the POMC gene will greatly aid our understanding of the precise molecular mechanisms controlling opioid peptide expression. PMID- 3028426 TI - Polyphosphoinositide metabolism in canine tracheal smooth muscle (CTSM) in response to a cholinergic stimulus. PMID- 3028428 TI - The specificity of omeprazole as an (H+ + K+)-ATPase inhibitor depends upon the means of its activation. AB - Omeprazole (OME) is a novel acid secretion inhibitor, believed to act directly on the gastric proton pump, the (H+ + K+)-ATPase. Inhibition of ATPase activity is associated with an incorporation of [14C]OME into gastric vesicles containing the (H+ + K+)-ATPase, and both processes are greatly enhanced if the OME is exposed to acidic pH. This, and other evidence, suggests that the acidic environment of the (H+ + K+)-ATPase generates from OME a reactive intermediate which covalently inhibits the pump. We have compared the means by which the OME was acid-activated with the specificity of inhibition (amount of incorporation of omeprazole required to produce 100% inhibition of K+-stimulated ATPase activity). The stoichiometry of incorporation has been related to the number of detectable catalytic phosphorylation sites in each preparation (an index of the number of functional pumps). In lyophilised gastric vesicles, where the membrane barriers separating the cytoplasmic and luminal faces of the enzyme are substantially destroyed, incubation with OME at pH 6.1 produced a progressive inhibition and incorporation over 120 min. Complete inhibition of K+-ATPase required 13 +/- 3 (SEM; N = 4) moles of OME incorporation per phosphorylation site. In intact gastric vesicles, under conditions shown independently to result in proton pumping and the acidification of the vesicle interior (150 mM KCl, 9 microM valinomycin, 2 mM Mg-ATP pH 7.0), inhibition and incorporation occurred more rapidly (15 min). Complete inhibition of K+-ATPase required only 1.8 +/- 0.15 (SEM; N = 3) moles of OME per phosphorylation site.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3028427 TI - Antiviral activity of 2',3'-dideoxycytidin-2'-ene (2',3'-dideoxy-2',3' didehydrocytidine) against human immunodeficiency virus in vitro. PMID- 3028430 TI - [Variants of phage M13 DNA containing a fragment of the beta-galactosidase gene- a convenient mutation system for the study of oligonucleotide-directed mutagenesis]. AB - A model system is developed to test oligonucleotide-directed mutations: T----C transition, T and C deletions (delta T and delta C), C insertion, double mutations (A----G, delta T), (T----C, A----G), and large oligonucleotide deletions (36 or 44 nucleotides). The system includes 9 variants of the phage M13 DNA carrying fragment of beta-galactosidase gene, and oligodeoxyribonucleotides partially noncomplementary to DNA sequence of this gene. Six variants are obtained by the site-localized mutagenesis, the other were described earlier. Induced mutations are easily tested by phenotype change of transformed bacteria (Lac+----Lac-); by formation or loss of the sites for BamHI and EcoRI restrictases; by DNA hybridization with 32P-labeled oligonucleotides; and by DNA sequencing by the Sanger method. The system is used to study the role of some factors, such as completeness of RF DNA synthesis, thermal stability of the oligonucleotide: DNA complex, quality of enzymes and substrates used in polymerase reaction, mutation type or the efficiency of mutagenesis. A number of unexpected mutations were observed in the course of oligonucleotide-directed mutagenesis. Lower yields of some mutants induced by oligonucleotides are shown to be due to the action of repair systems of bacteria. PMID- 3028429 TI - [Expression of the gene for hybrid beta-lactamase pBR322 with C-terminal fragment containing tuftsin (Thr-Lys-Pro-Arg)]. AB - A hybrid beta-lactamase gene with a synthetic tuftsin-coding DNA fragment inserted at the Pst I-site of pBR322 plasmid has been obtained and its expression has been studied. Radioactive amino acids have been used to show that in E. coli chi 925 minicells up to 30% of newly synthesized chimeric protein is secreted into periplasm providing the tuftsin transport. After hybrid protein cleavage with CNBr, tuftsin has been isolated using ion-exchange and thin-layer chromatography. PMID- 3028431 TI - [Synthesis of leukotriene A4 methyl ester via acetylene intermediates]. AB - Rac-leukotriene A4 methyl ester has been synthesized from propargylic alcohol and 1-heptyne. The synthetic strategy involves the assembly of carbon chain by acetylenide anion condensations and the introduction of (Z)-double bonds by the triple bond hydrogenation. PMID- 3028432 TI - Cytomegalovirus retinitis in rheumatic disease: a case report. AB - We describe a 67-year-old white man with Wegener's granulomatosis who was treated with cyclophosphamide, then with azathioprine, and who subsequently developed cytomegalovirus retinitis. The patient initially improved on a course of adenine arabinosine, but his retinitis has progressed, despite discontinuation of immunosuppressive therapy. We show that cytomegalovirus retinitis does occur in patients with rheumatic diseases who receive immunosuppressive therapy. PMID- 3028433 TI - The effect of urapidil on cardiac alpha- and beta-adrenoceptors. AB - In the present study the effects of the antihypertensive drug urapidil on cardiac adrenoceptors in various regions of the heart were analyzed. On the isolated rabbit papillary muscle urapidil was found to produce a competitive antagonism against the alpha 1-agonist phenylephrine with a pA2-value of 6.4 in the presence of the beta-antagonist sotalol (5 X 10(-5) mol/l). In the same preparation urapidil also antagonized the positive inotropic effects evoked by the beta adrenoceptor agonist isoprenaline yielding a pA2-value of 5.9. An antagonism against the positive inotropic effect of isoprenaline with the same pA2-value could also be demonstrated in guinea pig left atria. For further characterization of urapidil its effect against the beta 2-adrenoceptor agonist fenoterol on the carbachol contracted tracheal chain of the guinea pig was investigated. A weak antagonism with a pA2-value of 4.9 was observed. Thus the beta 1-adrenolytic potency of urapidil was found to be about 10 times higher than the beta 2 adrenolytic one. On the isolated perfused rabbit heart urapidil inhibited the electrically evoked increase in heart rate by 27 and 63% in a concentration of 10(-6) and 10(-5) mol/l, respectively, whereas the stimulation induced noradrenaline release was increased by 25 and 51%. This rise in noradrenaline release by urapidil was antagonized by clonidine 10(-7) and 10(-6) mol/l. From these results it can be concluded that urapidil acts not only as an antagonist at cardiac alpha 1- and beta-adrenoceptors, but also on alpha 2-adrenoceptors located presynaptically. The antagonism at these receptors against endogenous noradrenaline leads to an enhanced transmitter release.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3028434 TI - Protective effect of Nocardia rubra cell wall skeleton on experimental infection in normal and immunosuppressed mice. AB - The protective effect of Nocardia rubra cell wall skeleton (N-CWS) on experimental infections was investigated in normal and immunosuppressed mice. Pretreatment with N-CWS provided protection against acute systemic infections due to Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae, Serratia marcescens and Listeria monocytogenes in normal mice. N-CWS administered before or after challenge also showed protective activity against Herpes simplex virus infection in normal mice. N-CWS was the most effective against extracellular parasitic Pseudomonas aeruginosa infection when administered 1 day before challenge, but displayed the most potent protective activity against infection with Listeria monocytogenes, a facultative intracellular parasite, when administered 7 to 14 days before challenge. In mice with subcutaneous Pseudomonas aeruginosa infection, N-CWS suppressed abscess formation and decreased the viable cell count in the resultant abscess foci when administered either intraperitoneally or subcutaneously to the infected site. In addition, treatment with N-CWS markedly restored host defense ability against pseudomonal infection in mice immunosuppressed with cyclophosphamide, hydrocortisone and X-ray irradiation. PMID- 3028436 TI - Influence of the new angiotensin converting enzyme inhibitor ramipril on several models of acute inflammation and the adjuvant arthritis in the rat. AB - Paw oedema in the rat by carrageenin and kaolin partially caused by Hageman factor activation was potentiated by the new angiotensin converting enzyme (ACE) inhibitor 2-[N-[(S)-1-ethoxycarbonyl-3-phenylpropyl-L-alanyl]-(lS,3S,5S)-2- azabicyclo[3.3.0]octane-3-carboxylic acid] (ramipril, Hoe 498) due to its inhibition of kininase II which results in increased bradykinin levels. A dose of 1 microgram ramipril injected into the hind paw of Sprague-Dawley rats concomitantly with, or 1 mg/kg given orally 30 min before administration of the irritants, led to significantly increased inflammatory reactions. The same effects were observed when ramipril was administered 3 h after carrageenin. In the kallikrein-kinin-deficient Brown-Norway rat strain Mai Pfd/f, ramipril did not significantly alter the paw oedema induced as described above. In addition, pretreatment of Sprague-Dawley rats with 10 mg/kg i.v. bromelains completely prevented the potentiation of inflammation by ramipril. Paw oedema provoked by the Hageman factor non-activators serotonin, dextran, ovalbumin and anti-rat IgG was not potentiated by ramipril. The chronic adjuvant arthritis in Lewis rats was not influenced by daily oral treatment with 0.1-3 mg/kg ramipril. Thus, in the rat only those inflammatory reactions involving kinins, presumably generated by Hageman factor activators, are potentiated by ramipril and presumably by other ACE-inhibitors. PMID- 3028435 TI - Mespirenone and other 15,16-methylene-17-spirolactones, a new type of steroidal aldosterone antagonists. AB - The ability of a series of 15,16-methylene-spirolactones in comparison to known antimineralocorticoids to inhibit the renal actions of aldosterone was tested in adrenalectomized, glucocorticoid-treated rats. The standard procedure involved continuous i.v. infusion with an isotonic solution of low sodium content (0.05% NaCl + 5.2% glucose, 3 ml/rat/h) supplemented with d-aldosterone [1 micrograms/(kg X h)] resulting in a long-lasting reduction of renal sodium excretion, increase of renal potassium excretion and hence decrease of the urinary Na/K-ratio. In some experiments sodium input was increased (0.2% NaCl + 4.3% glucose or 0.9% NaCl, respectively). The test drugs either were administered orally 1 h before start of the infusion or were added to the infused solution. With the exception of two steroids which could only be tested at single doses, all compounds were administered at three doses ranging from 2.2 to 40 mg/kg (p.o.) or from 0.83 to 6.7 mg/kg/h (i.v.). Spironolactone or spirorenone (oral administration) and potassium canrenoate (i.v. infusion) served as reference compounds. The antimineralocorticoid activity of the steroids was judged by the increase in the aldosterone-lowered Na/K-ratio in urine which was collected at hourly intervals for 15 or 21 h, respectively. Adrenalectomized, glucocorticoid treated rats receiving an i.v. infusion without aldosterone were used as controls. To obtain preliminary information on potential antiandrogenic and progestogenic (side-)effects, binding of the test-compounds to androgen receptors (rat prostate cytosol) and progestogen receptors (rabbit uterus cytosol) was measured in vitro using 3H-dihydrotestosterone (DHT) and 3H-progesterone (prog.) as tracer and unlabelled DHT and prog. as references. All steroids tested exhibited antimineralocorticoid activity. For compounds tested at three doses levels the potency relative to the standard used was evaluated using regression analysis based on the Na/K-ratio or the log (Na X 100)/K-ratio. The relative potency of the other compounds was estimated by comparing the biological effect of single doses of test drug and standard drug, respectively, using nonparametric statistical tests.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3028437 TI - In vivo activation of functional properties in mouse peritoneal macrophages by Nocardia rubra cell wall skeleton. AB - Exudative cells in the peritoneal cavity of mice, particularly polymorphonuclear leukocytes significantly increased 1 day after intraperitoneal injection of Nocardia rubra cell wall skeleton (N-CWS). Macrophages and lymphocytes significantly increased 4 to 7 days after injection. N-CWS also enhanced peritoneal macrophage functions such as phagocytosis of latex particles, production of superoxide anion, production and secretion of lysosomal enzymes such as beta-glucuronidase, lysozyme and acid phosphatase, phagocytosis and intracellular killing of bacteria, and in vitro chemotaxis. The phagocytic function of the reticuloendothelial system was also enhanced. These results indicate that macrophages were activated in vivo by N-CWS. PMID- 3028438 TI - Humoral and blood pressure effects of the angiotensin converting enzyme inhibitor ramipril in essential hypertension. AB - The humoral and antihypertensive activities of the angiotensin converting enzyme (ACE) inhibitor 2-[N-[(S)-1-ethoxycarbonyl-3-phenylpropyl]-L-alanyl]-(1S, 3S, 5S) 2-azabicyclo[3.3.0] octane-3-carboxylic acid (ramipril, Hoe 498) were investigated in 10 patients with essential hypertension (WHO stage I or II). After a 7-day placebo period, the patients were treated with 5 mg ramipril orally once daily for 14 days. Peak serum concentrations of the active metabolite M1 (dicarboxylic acid) of 5.4-62.0 ng/ml were observed 2-6 h after the first oral dose. The maximum ACE inhibition of 95% was reached 2-4 h after the first oral dose, inhibition exceeded 70% 24 h after dosing. The maximum drop in the systolic and diastolic blood pressure (random zero sphygmomanometer) was measured 4 h after ramipril (p less than 0.02, p less than 0.01), but blood pressure on days 7 and 14 of the treatment period was not different from pretreatment values. Automatically recorded blood pressure results showed a marked reduction of both systolic and diastolic blood pressure during treatment compared to placebo. No side effects occurred. From the present data it is concluded that ramipril is a potent ACE inhibitor in hypertensive patients and that further controlled studies are required for the evaluation of the antihypertensive effect of 5 mg ramipril in essential hypertension. PMID- 3028439 TI - Peripheral neuropathy and myopathy in chronic alcoholism. AB - Nineteen chronic alcoholics referred for investigation of their medical complications and for alcohol re-education were subjected to quadriceps muscle biopsy and detailed peripheral nerve electrophysiological studies. Thirteen patients showed type II muscle fibre atrophy by histomorphometric analysis. The presence of atrophy did not correlate with peripheral neuropathy as determined by clinical examination or with objective indices of nutritional status. There was no association of muscle atrophy with neurophysiological evidence for neuropathy and both occurred independently of each other. It was concluded that chronic alcoholic myopathy was due to a toxic effect of ethanol on the muscle fibres and was not merely secondary to a peripheral neuropathy. PMID- 3028440 TI - Regional distribution of zinc, copper and lead in brain: (Na+-K+)-adenosine triphosphatase correlates of ethanol administration. AB - Zinc, copper and lead are known as inhibitory trace metals against brain (Na+-K+) ATPase. Alcohol (4 g/kg intraperitoneally for 10 days, to rats) induced an elevated level of lead in the cerebral cortex and cerebellum, whereas that of zinc was elevated only in latter region. Copper levels were found to be decreased in the hippocampus, amygdala and hypothalamus, but increased in the spinal cord. Zinc and lead contents were decreased in the amygdala and hypothalamus. The activity of (Na+-K+)-ATPase was enhanced in the hippocampus, amygdala and hypothalamus, but inhibited in the cerebral cortex and cerebellum. It is suggested that alcohol acts differentially on brain zinc, copper and lead concentrations and (Na+-K+)-ATPase activity. PMID- 3028441 TI - Biologic effects of chronic ethanol consumption related to a deficient intake of carbohydrates. AB - Our present understanding of the many effects of chronic ethanol consumption originated mostly from the use of an experimental model in which animals were maintained on a liquid diet that provided 36% of the total calories as alcohol. Since this diet was considered to be nutritionally adequate, a concept developed that the observed effects were due to the toxicity of alcohol and its metabolites rather than malnutrition. However, several biologic effects can be attributed to a reduced carbohydrate ingestion that accompanies chronic alcohol consumption. PMID- 3028442 TI - Loop diuretics causing reduction of voluntary alcohol drinking: relevance of thromboxane inhibition and opiate receptor activation? PMID- 3028444 TI - Physiological role of dietary fiber: a ten-year review. PMID- 3028443 TI - Treatment of disseminated cytomegalovirus infection with 9-(1,3 dihydroxy-2 propoxymethyl)guanine: evidence of prolonged survival in patients with the acquired immunodeficiency syndrome. AB - Disseminated cytomegalovirus (CMV) infection is a common complication of the acquired immunodeficiency syndrome (AIDS) and contributes significantly to its morbidity and mortality. Dihydroxypropoxymethyl guanine, DHPG, is an antiviral agent that has been shown to inhibit CMV replication and to provide clinical benefit in patients with CMV infections, especially retinitis. In this study, the clinical characteristics, results of diagnostic evaluations, and survival were compared in 11 AIDS patients with disseminated CMV infections who were seen between August 1981 and October 1984 and were not treated with DHPG, and in 18 AIDS patients seen since that time who were treated with DHPG. The study groups were similar though the untreated group was somewhat more tissue depleted. Survival from diagnosis was significantly prolonged with DHPG therapy based upon life table analysis (p = 0.001). Therapy improved the quality of life, as 12 of 18 treated patients and only 2 of 11 untreated patients could be discharged from the hospital. Progression of CMV infection did not appear to play a role in the mortality of patients who died during DHPG therapy. We conclude that DHPG prolongs survival in patients with AIDS who have disseminated CMV infections. PMID- 3028445 TI - Classification of lung carcinoma by means of digital nuclear image analysis. AB - An investigation was performed of the maximum discriminating efficiency for each subgroup of digital nuclear image features and of the overall classification of nuclei from three types of human lung carcinomas in histologic sections: adenocarcinoma, small-cell carcinoma and squamous-cell carcinoma. The results indicate that, for each subgroup of features, the nuclei of the small-cell carcinomas are generally "correctly" classified in a higher percentage (80% to 100%) than are the nuclei of the adenocarcinomas (46% to 74%) and squamous-cell carcinomas (29% to 68%). The discriminant analysis for the overall classification selected features from most of the subgroups, suggesting that it is useful to perform nuclear image analysis with many subgroups having different properties. The overall classifications for the nuclei of the adenocarcinomas, small-cell carcinomas and squamous-cell carcinomas were, respectively, 81.4%, 93.2% and 74.7%. Before this technique can be applied to histopathologic diagnosis, a larger number of unselected lung carcinomas must be evaluated. PMID- 3028446 TI - Mild but prolonged elevation of serum angiotensin converting enzyme (ACE) activity in alcoholics. AB - Serum activity of angiotensin converting enzyme (ACE) was serially measured in 47 hospitalized chronic alcoholics with liver disease. Compared to healthy controls, ACE activity, on admission, in the serum of alcoholics was significantly elevated (42.5 +/- 16.6 U/ml vs. 32.4 +/- 9.6 U/ml; p less than 0.005). About 36% of the patients had an elevated ACE level exceeding an upper normal value of 42 U/ml (mean +/- SD). In contrast to the rapid normalization of such enzymes as aspartate transaminase (AST), alanine transaminase (ALT) and lactic dehydrogenase (LDH) which represent parenchymal liver cell injury, the activity of ACE remained elevated over a period of 4 weeks even with abstinence. The serum level of ACE was significantly correlated with levels of alkaline phosphatase, gamma glutamyltranspeptidase and monoamine oxidase, but not with those of AST, ALT and LDH. These data suggest increased ACE activity in alcoholics may be related to the influence of chronic consumption of alcohol on hepatic nonparenchymal systems. PMID- 3028447 TI - [The quantitative determination of vitamin D3 and its metabolites in plasma]. AB - A method is described which enables determination of vitamin D3 and its physiologically most important metabolites, i.e. 25-OHD3, 24,25-(OH)2D3, 25,26 (OH)2D3 and 1,25-(OH)2D3 in a plasma sample of about 2 to 4 ml. The whole procedure involves two preparative and one analytical steps: Extraction with methanol/methylene chloride (2:1), chromatographic separation on Lipidex 5000 using a stepwise gradient of n-hexane and chloroform and finally HPLC separation on Zorbax-Sil columns with n-hexane isopropanol mixtures and subsequently reversed phase separation on RP 18-columns and mixtures of methanol and water. Except for 1,25-(OH)2D3 all D compounds were quantified by UV-detection with 1.4 ng of substance being the lowest detectable amount. 1,25-(OH)2D3 was measured by radioimmunoassay. Prior to HPLC analysis the extract was separated into three fractions on Lipidex 5000 which contained 1) vitamin D3, 2) 25-OHD3 and 3) the dihydroxy metabolites. The three fractions were separated by HPLC using different mixtures of isopropanol/n-hexane and methanol/water, respectively. Retention times of the individual D-components longer than 10 min appeared to be essential to separate these compounds from accompanying material. Overall recoveries of the individual metabolites were for vitamin D3 48.9%, for 25-OHD3 54.2%, for 24,25 (OH)2D3 50.9% and for 1,25-(OH)2D3 52.5%. Application of the methods to plasma samples from pigs with pseudovitamin D deficiency rickets, typ I, revealed a reduced concentration of 1,25-(OH)2D3 and 24,25-(OH)2D3 and an elevated level of 25-OHD3 in these animals. The results obtained by this method contributed substantially to a better understanding of the aetiological factors associated with this disease. PMID- 3028448 TI - Enhancer-like stimulation of yeast tRNA gene expression by a defined region of the Ty element micro-injected into Xenopus oocytes. AB - In the yeast Saccharomyces cerevisiae, the majority of the tRNA genes are found associated with transposable elements one of which is the Ty element. We noticed that the transcriptional activity of several of these genes was rather different depending on the type of the accompanying element(s) [Nelbock et al. (1985) Biol. Chem. Hoppe-Seyler 366, 1041-1051]. In order to study the influence of a Ty element on tRNA gene expression, we used the method of micro-injection into Xenopus oocytes taking advantage of a Ty+/Ty- allelic pair of a tRNALys1 gene recently isolated in our laboratory. A direct comparison showed that the expression of the plus allele was c. sixfold higher than that of the minus allele. In order to determine sequences of the Ty responsible for this effect, we constructed a number of variants in which distinct segments of the Ty were deleted or rearranged. The activating ability was localized to a 590-bp segment of the Ty containing the 'promoter delta', independent of its orientation or location towards the tRNA gene thus resembling the effects described for transcriptional enhancers. This is the first time that a long-range effect has been observed on the expression of a gene transcribed by RNA polymerase III. PMID- 3028449 TI - Protein-chemical identification of the major cleavage sites of the Ca2+ proteinase on murine vimentin, the mesenchymal intermediate filament protein. AB - Neutral thiol proteinases (calpains), activated by calcium are involved in the intracellular turnover of intermediate filaments but the precise position of the cleavage points has remained unknown. Here we identify by direct sequence analysis the major cleavage sites found when murine vimentin is digested by limited proteolysis in vitro with calpain purified from porcine kidney. Contrary to some previous suggestions, no absolute sequence specifity could be detected although 10 specific sites have been identified. This result is in line with the cDNA derived amino-acid sequence of a calpain, which pointed to a similarity of the catalytic site with the active sites in papain, cathepsin and actinidin. However, all major cleavage sites are located within regions of the vimentin molecule, which in current models of intermediate filament structure are thought to be non-helical: the amino-terminal headpiece, the carboxy-terminal tailpiece and the spacer separating the two major coiled-coil domains. The sequence information about the cleavage sites was extended to provide the amino-terminal 119 residues of murine vimentin. PMID- 3028451 TI - Synovial sarcoma of the head and neck. AB - Synovial sarcoma, a malignant soft-tissue tumor that occurs primarily in the extremities, is seen rarely in the head and neck. Although known to behave aggressively in the extremities, recent reports suggest that the course of synovial sarcoma may be more indolent when it occurs in the head and neck. Five new cases of synovial sarcoma of the head and neck are described herein. Of four patients with adequate follow-up, three died of pulmonary metastases at 1, 2 1/2, and 8 years following initial treatment. The fourth patient is alive with disease at the primary site six years after diagnosis. The poor prognosis for patients with this disease suggests that aggressive treatment is necessary. PMID- 3028450 TI - Production and characterization of two monoclonal antibodies against human myeloperoxidase. AB - Two monoclonal antibodies against human myeloperoxidase, designated 3-2H3 and 4 2C11, were produced and characterized. Both bound to the native enzyme, but neither bound to the denatured enzyme or to its two denatured subunits. 4-2C11 bound to the three types of leukocyte myeloperoxidase, I, II, and III, as well as to the four types of myeloid leukemia HL-60 cell myeloperoxidase, IA, IB, II, and III. 3-2H3 did not bind to enzyme IB, but bound to the other types of leukocyte and HL-60 cell enzymes. On incubation with myeloperoxidase III, 4-2C11 inhibited the enzyme activity, but 3-2H3 did not. Both antibodies belong to the IgG1 subclass. PMID- 3028452 TI - [n-Hexane polyneuropathy due to sniffing bond G10--clinical and electron microscopic findings]. AB - Two cases of n-hexane polyneuropathy due to glue sniffing were reported and the clinical features, laboratory data and sural nerve biopsy findings were discussed. Case 1 was a 31-year-old man and case 2 was a 24-year-old man. Both of them developed motor dominant polyneuropathy subacutely after inhalating Bond G10 for about 3 months. Their weakness deteriorated further for about 3 months even after discontinuing Bond G10 inhalation and they became almost confined to wheel chairs. Thereafter, however, they started to recover fairly rapidly. Gas chromatographic analysis of Bond G10 revealed that it contains 47.7% of n-hexane and 21.8% of toluene. Almost all reported cases of glue sniffer's neuropathy were motor dominant type, but those of n-hexane polyneuropathy of factory workers were sensory or sensorimotor neuropathy. Therefore our cases are thought to be typical of glue sniffer's neuropathies. Sural nerve biopsies were done in both patients, and examinations of photomicroscope, teased method and electronmicroscope were performed. From our photomicroscopic examinations we found only mild changes such as reduction of number of myelinated fibers, and from our electronmicroscopic examinations we noted marked accumulation of neurofilaments and loss of neurotubules in swelled axons. Some axons were normal in appearance by photomicroscopic examination, but the same axons under electronmicroscopic examination further revealed that neurotubules were oppressed by accumulation of neurofilaments to the margin of an axon. These findings seem to indicate the pathological processes of axonal swellings.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3028453 TI - [Pituitary adenoma calcification]. AB - Three hundred and eighteen cases of functioning and non-functioning pituitary adenoma were examined by histological, immunocytochemical and electronmicroscopic technique. Fourty-four of them (13.8%) showed evidence of calcospherites in the tumor tissues. A high incidence of calcospherite is found in functioning adenoma, but not in non-functioning adenoma. Calcification was seen most frequently in cases of prolactinoma (23), GH secreting (7), or GH + PRL tumor (2) and less in adrenocorticotropic hormone secreting adenoma (2) and follicle stimulating hormone secreting adenoma (1). Prolactin and growth hormone might be involved in the control of calcium metabolism. This is because, following adenomectomy in patients with prolactinoma or GH-secreting adenoma with hypercalcemia, there is normalization of serum PRL and GH with reduction in serum calcium. Calcospherite is produced in all of metastatic calcification, arterial calcification, dystrophic calcification and calcinosis. In cases of non-functioning adenoma however, the mechanism is believed to by dystrophic calcification. PMID- 3028455 TI - The acute haemodynamic effects of intravenous enalaprilic acid (MK422) in patients with left ventricular dysfunction. AB - Intravenous enalaprilic acid (2.5 mg) was given to 11 patients with stable cardiac failure (NYHA functional class II-IV). Reductions in mean right atrial, pulmonary artery and pulmonary capillary wedge pressure of 25%, 18% and 30% respectively (P less than 0.01), were observed. Cardiac output rose by 13% (NS) and mean blood pressure fell by 20% (P less than 0.01) with a decrease in systemic vascular resistance of 24% (P less than 0.01). Heart rate was unaltered. The haemodynamic effects correlated with control plasma renin activity (r = 0.78, P less than 0.01). Marked hypotension occurred in several subjects but no other side-effects were noted. The rapid onset of action and mixed venous and arteriolar dilating activity of intravenous enalaprilic acid may be an advantage in some clinical situations where parenteral vasodilating therapy is required. PMID- 3028454 TI - Detection of sputum eicosanoids in cystic fibrosis and in normal saliva by bioassay and radioimmunoassay. AB - We have measured arachidonic acid (AA) metabolites, leukotrienes (LTs) and prostanoids (Ps), in sputum of patients with cystic fibrosis (CF) and in normal saliva using bioassay and radioimmunoassay (RIA). Almost three times as much LTB4 is present in CF extracts compared with slow reacting substances (SRSs). Leukotrienes were not detected in normal saliva. In CF sputum there is a three fold increase in the level of the vasodilator prostanoid prostaglandin E2 (PGE2) and the stable metabolite of prostacyclin, 6-oxo PGF1 alpha compared with the vasoconstrictor prostaglandin F2 alpha (PGF2 alpha) and thromboxane B2 (TxB2), a hydrolysis product of thromboxane A2. Experiments with BW755c (25 micrograms ml 1, n = 3) indicated that the majority of this activity was not produced during the extraction procedure. The detection of LTs and Ps in sputum of CF patients shows that these substances are present at biologically active concentrations and may contribute to the pathophysiology of this disease. PMID- 3028456 TI - Absence of a pharmacokinetic interaction between enalapril and frusemide. AB - Single doses of enalapril maleate 10 mg and frusemide 80 mg do not significantly affect the pharmacokinetics of each other when taken concomitantly. Their concomitant use may be associated with more adverse effects than with the individual entities. PMID- 3028457 TI - Effect of enalapril on the skin response to bradykinin in man. AB - We tested the effect of oral enalapril on intradermal bradykinin to determine if kininase II inhibition occurs with therapeutic doses in vivo. Six normal male volunteers took either 5 mg enalapril orally or placebo on 2 days. Three hours later bradykinin was injected into the skin of the back in doses increasing from 10(-11) to 10(-9) M. Enalapril increased the bradykinin-induced wheal. Inhibition of kininase II may cause accumulation of endogenous bradykinin. This could be an important mechanism in the occasionally reported side effect of angioedema with the angiotensin converting enzyme (ACE) inhibiting group of drugs. PMID- 3028458 TI - The effect of an angiotensin converting enzyme inhibitor, ramipril, on bronchial responses to inhaled histamine and bradykinin in asthmatic subjects. AB - The effect of a potent inhibitor of angiotensin-converting enzyme, ramipril, was studied on both inhaled histamine and bradykinin-induced bronchoconstriction in six male, normotensive, mild asthmatic subjects. Oral administration of 10 mg ramipril caused no change in lung function or airway reactivity to inhaled histamine or bradykinin despite achieving adequate reduction in angiotensin converting enzyme activity. PMID- 3028459 TI - Binding of urokinase to specific receptor sites on human breast cancer membranes. AB - The high molecular weight form of the plasminogen activator urokinase (54 kD) binds to specific receptor sites on the cell membrane of breast carcinomas by its inactive "A" chain. The binding is of high affinity (range of dissociation constants: 5.6 X 10(-11) to 4 X 10(-10) mol l-1 and there were between 20 to 250 fmol of binding sites per milligram of membrane protein) and equilibrium is reached in 60 min. No competition for binding sites was observed with epidermal growth factor, tissue plasminogen activator or the low molecular weight form of urokinase (33 kD). Cross-linking experiments suggest that the receptor is a monomeric unit of molecular weight of 50 kD. This binding site provides a mechanism for the incorporation of urokinase into the cell membrane. PMID- 3028460 TI - The anti-proliferative effect of lithium chloride on melanoma cells and its reversion by myo-inositol. AB - The effect of LiCl on melanoma cell growth and differentiation was studied in mouse and human melanoma cell lines. LiCl markedly inhibited B16 and HT-144 melanoma cell growth in vitro. Clonogenicity in soft agar of the melanoma cells was also markedly inhibited by LiCl. Pretreatment of B16 mouse melanoma cells with LiCl delayed the appearance of melanoma tumours in syngeneic mice. Growth inhibition of cells was accompanied by morphological and biochemical alterations. LiCl induced cell enlargement and formation of dendrite-like structures. The activity of NADPH cytochrome c reductase, an enzymatic marker of endoplasmic reticulum was significantly (2-3 fold) increased. Addition of myo-inositol to cell cultures partially reversed the anti-proliferative and morphological effects of LiCl on melanoma cells. This finding may suggest that the anti-proliferative effect of LiCl is related to its effect on phosphatidylinositol metabolism. PMID- 3028462 TI - Placental-type alkaline phosphatase in cervical neoplasia. AB - Monoclonal antibodies reactive with placental-type alkaline phosphatase have formed the basis of methods for detection of this oncodevelopmental antigen in patients with pre-invasive and invasive cervical neoplasia, with or without evidence of papilloma virus infection. Disease-related elevations of placental type alkaline phosphatase were not observed in patients' sera. Solubilised cervical smears or biopsy material, and cervical mucus swabs, often contained substantial amounts of this isoenzyme; however, there was no significant difference between any of the patient and control groups. Thus, serological and smear test assays for placental-type alkaline phosphatase were not useful in differential diagnosis of cervical lesions. However, its presence in most biopsy specimens, often at high levels, indicated possible application for in vivo radioimmunoimaging studies of invasive or metastatic cervical cancer. PMID- 3028461 TI - HLA antigens in colorectal tumours--low expression of HLA class I antigens in mucinous colorectal carcinomas. AB - Expression of HLA antigens and beta 2-microglobulin was studied by immunoperoxidase staining of frozen sections of 9 mucinous and 10 nonmucinous colorectal adenocarcinomas, 1 cloacogenic carcinoma, 12 colorectal adenomas and 4 samples of normal colorectal mucosa using monoclonal antibodies (MAbs). Staining results were related to histopathological features. HLA Class I antigens were strongly expressed in morphologically normal colorectal epithelium, in all adenomas tested and in all non-mucinous carcinomas. In contrast, expression of HLA class I antigens by the majority of tumour cells was present in only 2 of the 9 mucinous carcinomas, whereas 2 of these mucinous carcinomas were completely negative. In the mucinous carcinomas a striking scarcity of mononuclear inflammatory infiltrate, especially around the mucus accumulations, was observed. HLA class II antigen expression was not detected in normal epithelium and was only focally present in 1 of the 12 adenomas. In 6 out of the 20 carcinomas tested between 20% and 90% of the tumour cells were stained by MAbs against HLA class II antigens. Apart from the low expression of HLA class I antigens in mucinous carcinomas no relationship was found between expression of HLA antigens and histological features of the tumours. The relative poor prognosis of mucinous colorectal carcinoma as reported in the literature may be associated with low expression of HLA class I antigens and scant mononuclear inflammatory infiltrate, which may be a reflection of a weak immune response to the tumour cells. PMID- 3028464 TI - Anti-oxidant effects of gold compounds. AB - The anti-oxidant efficacy, in vitro, of the gold compounds auranofin (AF) and gold sodium thiomalate (GST) was examined by studying their effects on the generation of reactive oxygen species (ROS) using zymosan-stimulated polymorphonuclear leukocytes (PMNs) and a cell-free, xanthine-xanthine oxidase system. The oxygen species investigated were the superoxide radical anion (O2-), hydrogen peroxide (H2O2) and the hydroxyl radical (OH.). AF had an inhibitory effect on ROS production by PMNs. In particular, OH. generation was significantly suppressed in a dose-dependent fashion. AF did not inhibit ROS production in the cell-free system. GST produced only a small degree of inhibition at higher concentrations. These findings suggest that AF may play an important role in the inhibition of respiratory bursts and the generation of inflammatory reaction products. Since the products of the respiratory burst, especially potent oxidants such as OH. and H2O2, are thought to be important inflammatory mediators, it is postulated that the blockade of toxic ROS generation by AF affects rheumatoid as well as dermatological inflammation and tissue damage. PMID- 3028463 TI - Semliki Forest virus induced, immune mediated demyelination: the effect of irradiation. AB - Intraperitoneal infection with the avirulent A7(74) strain of the alphavirus Semliki Forest virus (SFV) induces an immune mediated demyelinating encephalomyelitis. The blood and brain virus titres, the serum antibody titres and the histopathological changes in the brains of normal mice and mice immunosuppressed with 5.0 or 8.0 Gy total body irradiation (TBX) were determined. SFV infection of immunosuppressed mice resulted in persistently high blood and brain virus titres, neuronal pycnosis, paralysis and death. No demyelination or central nervous system (CNS) inflammatory response occurred in these immunosuppressed mice despite high and persistent brain virus titres. The CNS inflammatory response and associated demyelination could be restored to infected immunosuppressed mice by adoptive transfer of spleen cells, and these changes were brought forward if the donor spleen cells were from mice previously sensitized to SFV. The results indicate that the immune response following SFV A7(74) infection is both protective and pathogenic, and that the demyelination is immune mediated and does not result from direct viral destruction of oligodendrocytes, or any other direct effect of the virus. PMID- 3028465 TI - Pagetoid reticulosis, epidermotropic mycosis fungoides and mycosis fungoides: a disease spectrum. AB - Using a standard technique involving monoclonal antibodies against T-cell subsets, we have shown that almost all the infiltrating T-cells in the epidermis of a patient with Pagetoid reticulosis (PR), one with epidermotropic mycosis fungoides (EMF) and one with poikiloderma atrophicans vasculare (PAV), were OKT8 positive (presumed cytotoxic/suppressor) T-cells. The infiltrating T-cells in the epidermis of a patient with limited plaque stage mycosis fungoides (MF), however, were almost exclusively Leu 3a-positive (presumed helper/inducer) T-cells as is usually found in this condition. The keratinocytes in the patients with PR, EMF and PAV were HLA-DR-positive whilst those in the patient with MF were HLA-DR negative. We consider these four diseases to be part of the spectrum of mycosis fungoides, the first three conditions representing the early or benign end of the spectrum. PMID- 3028466 TI - Chromatographic analysis of human erythrocyte pyrimidine 5'-nucleotidase from five patients with pyrimidine 5'-nucleotidase deficiency. AB - Human erythrocyte pyrimidine 5'-nucleotidase (P5N) was separated into two subclasses. P5N-I and P5N-II, by DEAE Bio-Gel A column chromatography. Their enzymological properties were studied using five normal subjects and five patients with different P5N deficiencies. Study of the normal subjects showed that P5N-I and P5N-II have distinctive properties, and P5N-II is similar to the 5'-nucleotidase in rat liver cytosol. The P5N-II from the five subjects with this deficiency had normal activity and other normal enzymological properties. However, the P5N-I from these patients had abnormal properties, including reduced activity. These variant enzymes had a high Michaelis constant for substrate cytidine 5'-monophosphate and were heat stable. The optimum pH was shifted towards the acidic side in two patients, towards the basic side in one, and was unchanged in the other two. These results strongly suggest that the main cause of P5N deficiency is an abnormality of P5N-I, probably arising from a structural gene mutation. PMID- 3028467 TI - Subpopulations of Langerhans' cells in cervical neoplasia. AB - Multiple markers were used to count Langerhans' cells in the cervix. In the normal cervix, thymocyte antigen (T6) and adenosine triphosphatase (ATPase) demonstrated the largest population of Langerhans' cells. MHC Class II positive cells were equivalent to 60%, and S100 positive cells were equivalent to 35% of T6 or ATPase positive cells. Whereas Langerhans' cells demonstrated by T6, ATPase, and MHC Class II antigen were evenly distributed throughout the epithelium, the S100 positive cells were seen predominantly near lymphocytic aggregates and capillaries. In human papillomavirus infection and cervical intraepithelial neoplasia the numbers of T6, ATPase, or MHC Class II positive Langerhans' cells were reduced by 60% but the S100 positive cells were almost completely depleted. These findings suggested that there were different subpopulations of Langerhans' cells in the cervical epithelium. The depletion of Langerhans' cells, particularly the selective depletion of the S100 positive subpopulation, might cause a localized immunodeficiency that impairs immune surveillance and the cell-mediated immune response to human papillomavirus infection and cervical intraepithelial neoplasia. PMID- 3028468 TI - Lymphocyte phenotypes in cervical intraepithelial neoplasia and human papillomavirus infection. AB - Lymphocyte phenotypes in cervical mucosa were studied using a panel of monoclonal antibodies. T lymphocytes were predominant both within the epithelium and in the subepithelial stroma. In the normal cervix, both the T4+ (helper/inducer) and T8+ (suppressor/cytotoxic) subsets were present in a ratio similar to that in the peripheral circulation. In human papillomavirus (HPV) infection and cervical intraepithelial neoplasia (CIN) there was depletion of intraepithelial lymphocytes, especially of T4+ subset, with reversal of the ratio of T4+ to T8+ subsets to less than one. In contrast, there was no significant reduction in the number of lymphocytes in the subepithelial stroma. Tac+ (antigen primed and clonal expanding) lymphocytes were absent both within the epithelium and in the subepithelial stroma. These findings support our suggestion that there is a localized immunodeficiency in HPV infection and CIN. The aetiological and therapeutic implications are discussed. PMID- 3028469 TI - Infiltrating orbital granular cell tumour: a case report and literature review. AB - A surgical biopsy of an infiltrative retrobulbar mass in a 44-year-old man was diagnosed as granular cell tumour. Electron microscopy and immunoperoxidase stains were used to confirm the diagnosis and to study the histogenesis of this rare soft tissue neoplasm. S-100 stain was positive, while neuron-specific enolase and myoglobin stains were negative, suggesting a non-specific neural origin for the cells. The capability of this tumour to invade surrounding tissues has seldom been described in the orbit and is demonstrated by this case. PMID- 3028470 TI - Molecular cloning of three distinct forms of the Na+,K+-ATPase alpha-subunit from rat brain. AB - Rat brain and kidney cDNA libraries were constructed and screened with a cDNA insert corresponding to the mRNA for the sheep kidney Na+,K+-ATPase catalytic subunit. The alpha-subunit cDNAs isolated from the kidney library were derived from a single class of messenger RNA, and the brain cDNAs were derived from three classes of messenger RNA. The most abundant brain cDNA, which spans 5.1 kilobases, encodes the alpha(+) form of the enzyme. The second most abundant brain cDNA, which spans 3.65 kilobases, is identical with that of the kidney form and therefore encodes the alpha isoform. The third class of cDNA, which spans 3.55 kilobases, was present at low abundance and encodes an isoform of the alpha subunit, designated alpha III, which has not been identified previously. The complete nucleotide sequence and deduced amino acid sequence for each of the brain and kidney cDNAs have been determined. In addition, we have identified a lysine-rich sequence that may function as a movable, ion-selective gate during cation binding and occlusion and have also identified several amino acid sequence variations that appear to explain some of the well-known species and tissue differences in cardiac glycoside sensitivity. PMID- 3028471 TI - Spatial relationship and conformational changes between the cardiac glycoside site and beta-subunit oligosaccharides in sodium plus potassium activated adenosinetriphosphatase. AB - (Na,K)-ATPase, the enzyme responsible for active transport of Na and K across the plasma membranes of animal cells, consists of a catalytic subunit (alpha) and a glycoprotein subunit (beta) with unknown function. We have determined the distance between fluorescent probes directed to specific sites on the alpha- and beta-subunits and ligand-induced changes in the fluorescence of a probe specifically attached to the beta-subunit. The cardiac glycoside site on the alpha-subunit was labeled with anthroylouabain [Fortes, P. A. G. (1977) Biochemistry 16, 531-540]. The oligosaccharides on the beta-subunit were labeled with lucifer yellow carbohydrazide [Lee, J. A., & Fortes, P. A. G. (1985) Biochemistry 24, 322-330]. Resonance energy transfer from anthroylouabain to lucifer yellow was measured by steady-state and time-resolved fluorescence spectroscopy. The distance between these probes was determined from the efficiency of energy transfer. The average distance between anthroylouabain and lucifer yellow was 47 A and was independent of the number of acceptor molecules attached to the beta-subunit. The measured distance corresponds to the distance between the cardiac glycoside site and the center of the labeled oligosaccharides on the beta-subunit within one alpha beta dimer. The distance was the same (47 A) when anthroylouabain was bound with ATP or Pi as phosphorylating ligands but increased to 49 A in the presence of vanadate. The change in average distance provides quantitative evidence of a conformational difference between the complexes of cardiac glycosides with (Na,K)-ATPase induced by phosphorylating ligands or by vanadate.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3028472 TI - Carbon-13 and deuterium isotope effects on oxalacetate decarboxylation by pyruvate carboxylase. AB - Deuterium and 13C isotope effects for the enzymic decarboxylation of oxalacetate showed that both deuterium- and 13C-sensitive steps in the reaction are partially rate limiting. A normal alpha-secondary effect of 1.2 per deuterium was calculated for the reaction in which pyruvate-d3 was the substrate, suggesting that the enolate of pyruvate was an intermediate in the reaction. The large normal alpha-secondary deuterium isotope effect of 1.7 when oxalacetate-d2 was the substrate suggests that the motions of the secondary hydrogens are coupled to that of the primary hydrogen during the protonation of the enolate of pyruvate. The reduction in the magnitude of the 13C isotope effect for the oxamate dependent decarboxylation of oxalacetate from 1.0238 to 1.0155 when the reaction was performed in D2O (primary deuterum isotope effect = 2.1) clearly indicates that the transfer of the proton and carboxyl group between biotin and pyruvate does not occur via a single concerted reaction. Mechanisms in which biotin is activated to react with CO2 (prior to transfer of the proton on N-1) by bond formation between the sulfur and the ureido carbon, or in which the sequence of events is decarboxylation of oxalacetate, proton transfer from biotin to enolpyruvate, and carboxylation of enolbiotin, predict that the 13C isotope effect in D2O should be substantially lower than the observed value. A stepwise mechanism that does fit the data is one in which a proton is removed from biotin by a sulfhydryl group on the enzyme prior to carboxyl transfer, as long as the sulfhydryl group has an abnormally low pK.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3028473 TI - Complete sequence and structure of the gene for human adenosine deaminase. AB - The nucleotide sequence of the human adenosine deaminase gene was determined. The gene was isolated in a series of overlapping lambda phage clones containing human germ line DNA. A total of 36,741 base pairs were sequenced, including 32,040 base pairs from the transcription initiation site to the polyadenylation site, 3935 base pairs of 5'-flanking DNA, and 766 base pairs of 3'-flanking DNA. The gene contains 12 exons separated by 11 introns. The exons range in size from 62 to 325 base pairs while the introns are 76-15 166 base pairs in size. The area sequenced contains 23 copies of Alu repetitive DNA and a single copy of an "O" family repeat. All but one of these repeat sequences are located in the first three introns or the 5'-flanking region. The apparent promoter region of the gene lacks the "TATA" and "CAAT" sequences often found in eucaryotic promoters and is extremely G/C rich. Contained within this region are areas homologous to other G/C-rich promoters, including six decanucleotide sequences that are highly homologous to sequences identified as functional binding sites for transcription factor Sp1. PMID- 3028474 TI - Parameters affecting fusion between Sendai virus and liposomes. Role of viral proteins, liposome composition, and pH. AB - A kinetic and quantitative characterization of the fusion process between Sendai virus and phospholipid vesicles is presented. Membrane fusion was monitored in a direct and continuous manner by employing an assay which relies on the relief of fluorescence self-quenching of the probe octadecylrhodamine B chloride which was located in the viral membrane. Viral fusion activity was strongly dependent on the vesicle lipid composition and was most efficient with vesicles solely consisting of acidic phospholipids, particularly cardiolipin (CL). This result implies that the fusion of viruses with liposomes does not display an absolute requirement for specific membrane receptors. Incorporation of phosphatidylcholine (PC), rather than phosphatidylethanolamine (PE), into CL bilayers strongly inhibited fusion, suggesting that repulsive hydration forces interfere with the close approach of viral and target membrane. Virus-liposome fusion products were capable of fusing with liposomes, but not with virus. In contrast to fusion with erythrocyte membranes, fusion between virus and acidic phospholipid vesicles was triggered immediately, did not strictly depend on viral protein conformation, and did not display a pH optimum around pH 7.5. On the other hand, with vesicles consisting of PC, PE, cholesterol, and the ganglioside GD1a, the virus resembled more closely the fusogenic properties that were seen with erythrocyte target membranes. Upon decreasing the pH below 5.0, the viral fusion activity increased dramatically. With acidic phospholipid vesicles, maximal activity was observed around pH 4.0, while with GD1a-containing zwitterionic vesicles the fusion activity continued to increase with decreasing pH down to values as low as 3.0.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3028475 TI - Mass action analysis of kinetics and extent of fusion between Sendai virus and phospholipid vesicles. AB - The kinetics and extent of fusion between Sendai virus particles and liposomes were investigated with an assay for lipid mixing based on the relief of self quenching of fluorescence. The measurements, which were carried out at pH 7.4 and 5.0, included liposomes of three compositions, cardiolipin (CL), CL/dioleoylphosphatidylcholine (CL/DOPC 1:1), and phosphatidylserine (PS). Liposomal lipid concentrations varied from 2.5 to 50 microM. In addition, the effect of low concentrations of the dehydrating agent poly(ethylene glycol) (PEG) on fusion between the virus and the liposomes at pH 7.4 was studied. The results were analyzed in terms of a mass action kinetic model which views the overall fusion reaction as a sequence of a second-order process of virus-liposome adhesion or aggregation, followed by the first-order fusion reaction itself. The fusion products were shown to consist of a single virus particle and several liposomes. Analytical solutions were found for the final extent of fusion and increase in fluorescence intensity following the fusion of fluorescently labeled virus particles with liposomes. The final extents of fluorescence intensity were explained by assuming an essentially irreversible binding of liposomes to inactive virus particles. The percents of active virus particles and the rate constants of fusion and aggregation were larger at pH 5 than at pH 7.4, increased when PEG was included in the medium, and varied with liposomal lipid composition according to the sequence CL greater than CL/DOPC greater than PS.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3028476 TI - S-adenosylhomocysteinase: mechanism of reversible and irreversible inactivation by ATP, cAMP, and 2'-deoxyadenosine. AB - Homogeneous S-adenosylhomocysteinase (AdoHcyase) from rat liver is a tetrameric enzyme that contains four molecules of tightly bound NAD per mole of enzyme. We report here that incubation of the rat liver enzyme with ATP, Mg2+, and KCl leads to conversion of the active enzyme to an inactive form with release of all enzyme bound NAD which can be recovered quantitatively by gel filtration. At various concentrations of ATP, the release of NAD corresponds closely with the degree of inactivation, suggesting that the four subunits are equivalent. Hydrolysis of ATP is not required for the inactivation process since nonhydrolyzable ATP analogues can replace ATP in the inactivation process. The ATP-dependent inactivation is fully reversible upon incubation of the inactivated enzyme with NAD. The ATP dependent inactivation of the enzyme appears to be analogues to the cAMP dependent inactivation of the enzyme from Dictyostelium discoideum described earlier by Hohman et al. (1985) [Hohman, R. J., Guitton, M. C., & Veron, M. (1985) Proc. Natl. Acad. Sci. U.S.A. 82, 4578-4581; Hohman, R. J., Veron, M., & Guitton, M. C. (1985) Curr. Top. Cell. Regul. 26, 233-245] but differs from the irreversible inactivation studied earlier by Abeles et al. (1982) [Abeles, R. H., Fish, S., & Lapinskas, B. (1982) Biochemistry 21, 5557-5562]. These authors have ascribed the time-dependent inactivation that results from incubation of the enzyme with 2'-deoxyadenosine at the C-3' and concluded that AdoHcyase "probably consists of two nonequivalent pairs of subunits".(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3028478 TI - The distance between cytochromes a and a3 in the azide compound of bovine-heart cytochrome oxidase. AB - The electron-spin relaxation rates of the two species of cytochrome a3(3+)-azide found in the azide compound of bovine-heart cytochrome oxidase were measured by progressive microwave saturation at T = 10 K. It has been shown previously that Cyt a3(3+)-azide gives rise to two distinct EPR resonances, depending upon the oxidation state of Cyt a. When Cyt a is ferrous, Cyt a3(3+)-azide has g = 2.88, 2.19 and 1.64; upon oxidation of Cyt a, the a3(3+)-azide g-values become g = 2.77, 2.18, and 1.74 (Goodman, G. (1984) J Biol. Chem. 259, 15094-15099). The relaxation effect of Cyt a on Cyt a3 could be measured as the difference in microwave field saturation parameter H1/2 between the g = 2.77 and g = 2.88 species. For each signal the spin-lattice relaxation time T1 was determined from H1/2 using the transverse relaxation time T2. The value of T2 at 10 K was extrapolated from a plot of line-width vs. temperature at higher temperature. The dipolar contribution to T1 was related to the Cyt a-Cyt a3 spin-spin distance utilizing available information on the relative orientation of Cyt a3-azide and Cyt a (Erecinska, M., Wilson, D.F. and Blasie, J.K. (1979) Biochim. Biophys. Acta 545, 352-364). By taking into account the relaxation parameters for both gx and gz components of the Cyt a3-azide g-tensor, the angle between the gz components of the Cyt a and Cyt a3 g-tensors was determined to be between 0 and 18 degrees, and the Cyt a-Cyt a3 spin-spin distance was found to be 19 +/- 8 A. PMID- 3028477 TI - Phospholipid-dependent interaction between dibromothymoquinone and iron-sulfur protein in mitochondrial ubiquinol-cytochrome c reductase. AB - Dibromothymoquinone (DBMIB) inhibits antimycin A-sensitive ubiquinol-cytochrome c reductase activity; the maximal inhibition is 90%. DBMIB alters the EPR spectra of reduced iron-sulfur protein in intact ubiquinol-cytochrome c reductase. The maximal spectral change occurs with 60 mol inhibitor per mol cytochrome c1 in the reductase. DBMIB causes little alteration in the EPR characteristics of iron sulfur protein when ubiquinol-cytochrome c reductase is delipidated. When delipidated ubiquinol-cytochrome c reductase is replenished with phospholipid, the effect of DBMIB reappears. However, when DBMIB is added to delipidated protein prior to replenishment with phospholipid, very little spectral alteration is observed. DBMIB does not alter the EPR spectra of purified iron-sulfur protein, with or without phospholipid in the preparation. Reduced DBMIB does not alter the EPR characteristics of iron-sulfur protein in intact or delipidated ubiquinol-cytochrome c reductase. Cysteine and other thiol compounds can reverse the spectral alternation caused by DBMIB. This reversal probably results from the reduction of DBMIB. PMID- 3028479 TI - Assignment of the g = 4.1 EPR signal to manganese in the S2 state of the photosynthetic oxygen-evolving complex: an X-ray absorption edge spectroscopy study. AB - X-ray absorption spectroscopy at the Mn K-edge has been utilized to study the origin of the g = 4.1 EPR signal associated with the Mn-containing photosynthetic O2-evolving complex. Formation of the g = 4.1 signal by illumination of Photosystem II preparations at 140 K is associated with a shift of the Mn edge inflection point to higher energy. This shift is similar to that observed upon formation of the S2 multiline EPR signal by 190 K illumination. The g = 4.1 signal is assigned to the Mn complex in the S2 state. PMID- 3028480 TI - Dynamic fluorescence quenching studies on lipid mobilities in phosphatidylcholine cholesterol membranes. AB - Bimolecular collision rate of 8-anilinonaphthalene-1-sulfonic acid (ANS) and the nitroxide doxyl group attached to various carbons on stearic acid spin labels (n SASL) in phosphatidylcholine-cholesterol membranes in the fluid phase was studied by observing dynamic quenching of ANS fluorescence by n-SASL's. The excited-state lifetime of ANS and its reduction by the n-SASL doxyl group were directly measured by the time-correlated single photon counting technique to observe only dynamic quenching separately from static quenching and were analyzed by using Stern-Volmer relations. The collision rate of ANS with the n-SASL doxyl group ranges between 1 X 10(7) and 6 X 10(7), and the extent of dynamic quenching by n SASL is in the order of 5-much much greater than 6- greater than 7- less than 9- less than 10- less than 12- less than 16-SASL (less than 5-SASL) in dimyristoylphosphatidylcholine (DMPC) membranes. Collision rate of 16-SASL is only 10% less than that of 5-SASL. Since the naphthalene ring of ANS is located in the near-surface region of the membrane, these results indicate that the methyl terminal of SASL appears in the near surface area frequently, probably due to extensive gauche-trans isomerism of the methylene chain. The presence of 30 mol% cholesterol decreases the collision rate of ANS with 12- and 16-SASL doxyl groups but not with the 5-SASL doxyl group in DMPC membranes. On the other hand, in egg-yolk phosphatidylcholine membranes, inclusion of 30 mol% cholesterol does not affect the collision of ANS with either 5-SASL or 16-SASL doxyl groups, in agreement with our previous observation that alkyl chain unsaturation moderates cholesterol effects on lipid motion in the membrane (Kusumi et al., Biochim. Biophys. Acta 854, 307-317). It is suggested that dynamic quenching of ANS fluorescence by lipid-type spin labels is a useful new monitor of membrane fluidity that reports on various lipid mobilities in the membrane; a class of motion can be preferentially observed over others by selecting a proper spin label, i.e., rotational diffusion of lipid about its long axis and translational diffusion by using 5-SASL, wobbling motion of the lipid long axis by using 7-SASL or androstane spin label, and gauche-trans isomerism by using 16-SASL. PMID- 3028482 TI - Stabilization of small unilamellar phospholipid vesicles during spray-drying. AB - In the presence of 10% (0.3 M) sucrose in the aqueous medium, small unilamellar phospholipid vesicles are preserved during freeze-drying and spray-drying. Moreover, the bilayer integrity and permeability barrier are maintained during these processes. PMID- 3028481 TI - (Na+ + K+)-ATPase: confirmation of the three-pool model for the phosphointermediates of Na+-ATPase activity. Estimation of the enzyme-ATP dissociation rate constant. AB - The dephosphorylation kinetics of acid-stable phosphointermediates of (Na+ + K+) ATPase from ox brain, ox kidney and pig kidney was studied at 0 degree C. Experiments performed on brain enzyme phosphorylated at 0 degree C in the presence of 20-600 mM Na+, 1 mM Mg2+ and 25 microM [gamma-32P]ATP show that irrespectively of the EP-pool composition, which is determined by Na+ concentration, all phosphoenzyme is either ADP- or K+-sensitive. After phosphorylation of kidney enzymes at 0 degree C with 1 mM Mg2+, 25 microM [gamma 32P]ATP and 150-1000 mM Na+ the amounts of ADP- and K+-sensitive phosphoenzymes were determined by addition of 1 mM ATP + 2.5 mM ADP or 1 mM ATP + 20 mM K+. Similarly to the previously reported results on brain enzyme, both types of dephosphorylation curves have a fast and a slow phase, so that also for kidney enzymes a slow decay of a part of the phosphoenzyme, up to 80% at 1000 mM Na+, after addition of 1 mM ATP + 20 mM K+ is observed. The results obtained with the kidney enzymes seem therefore to reinforce previous doubts about the role played by E1 approximately P(Na3) as intermediate of (Na+ + K+)-ATPase activity. Furthermore, for both kidney enzymes the sum of ADP- and K+-sensitive phosphoenzymes is greater than E tot. In experiments on brain enzyme an estimate of dissociation rate constant for the enzyme-ATP complex, k-1, is obtained. k-1 varies between 1 and 4 s-1 and seems to depend on the ligands present during formation of the complex. The highest values are found for enzyme-ATP complex formed in the presence of Na+ or Tris+. The results confirm the validity of the three-pool model in describing dephosphorylation kinetics of phosphointermediates of Na+-ATPase activity. PMID- 3028483 TI - Orientation and vertical fluctuations of spin-labeled analogues of cholesterol and androstanol in phospholipid bilayers. AB - We have used ESR and NMR linewidth broadening by spin-labels to determine the overall orientation of spin-labeled analogues of cholesterol and androstanol in egg lecithin bilayers. While the cholesterol analogues were found to have a single orientation in each monolayer, with the acyl chain pointing towards the center of the bilayer, the androstanol analogue appeared, at least in sonicated vesicles, to experience two opposite orientations in the same monolayer, very likely with a rapid reorientation. The possibility of rapid vertical fluctuations of the sterol molecules within the phospholipid bilayer is also discussed. PMID- 3028484 TI - A sphingomyelinase-resistant pool of sphingomyelin in the nuclear membrane of hen erythrocytes. AB - Experiments in which hen erythrocytes were exposed to the action of exogenous sphingomyelinase (Staphylococcus aureus) or to their endogenous plasma membrane sphingomyelinase showed that about 15% of the total sphingomyelin was resistant to breakdown either in intact or lysed cells. This resistant pool of sphingomyelin seems likely to reside in the nuclear membranes of the cells, so that essentially all the plasma membrane sphingomyelin can be broken down by exogenous sphingomyelinase acting on intact cells, suggesting that plasma membrane sphingomyelin is exclusively localised in the outer lipid leaflet. Paradoxically, introduction of Ca2+ into the intact cells using A23187 causes the breakdown of up to 30% of total cell sphingomyelin inside the cells but without apparently affecting the putative nuclear pool of sphingomyelin and this suggests that Ca2+ may alter the original disposition of sphingomyelin in the membrane so that originally outer leaflet sphingomyelin becomes accessible to the endogenous sphingomyelinase inside the cells. No differences were seen in the fatty acid compositions of sphingomyelin degradable by exogenous sphingomyelinase, sphingomyelin degradable in the presence of A23187/Ca2+ or the enzyme-resistant pool of sphingomyelin. PMID- 3028485 TI - Skeletal muscle sarcolemma in malignant hyperthermia: evidence for a defect in calcium regulation. AB - Sarcolemmal properties implicated in the skeletal muscle disorder, malignant hyperthermia (MH), were examined using sarcolemma-membrane vesicles isolated from normal and MH-susceptible (MHS) porcine skeletal muscle. MHS and normal sarcolemma did not differ in the distribution of the major proteins, cholesterol or phospholipid content, vesicle size and sidedness, (Na+ + K+)-ATPase activity, ouabain binding, or adenylate cyclase activity (total and isoproterenol sensitivity). The regulation of the initial rates of MHS and normal sarcolemmal ATP-dependent calcium transport (calcium uptake after 1 min) by Ca2+ (K1/2 = 0.64 0.81 microM), calmodulin, and cAMP-dependent protein kinase were similar. However, when sarcolemmal calcium content was measured at either 2 or 20 min after the initiation of active calcium transport, a significant difference between MHS and normal sarcolemmal calcium uptake became apparent, with MHS sarcolemma accumulating approximately 25% less calcium than normal sarcolemma. Calcium transport by MHS and normal sarcolemma, at 2 or 20 min, had a similar calmodulin dependence (C1/2 = 150 nM), and was stimulated to a similar extent by cAMP-dependent protein kinase or calmodulin. Halothane inhibited MHS and normal sarcolemmal active calcium uptake in a similar fashion (half-maximal inhibition at 10 mM halothane), while dantrolene (30 microM) and nitrendipine (1 microM) had little effect on either MHS or normal sarcolemmal calcium transport. After 20 min of ATP-supported calcium uptake, 2 mM EGTA plus 10 microM sodium orthovanadate were added to initiate sarcolemmal calcium efflux. Following an initial rapid phase of calcium release, an extended slow phase of calcium efflux (k = 0.012 min 1) was similar for both MHS and normal sarcolemma vesicles. We conclude that although a number of sarcolemmal properties, including passive calcium permeability, are normal in MH, a small but significant defect in MHS sarcolemmal ATP-dependent calcium transport may contribute to the abnormal calcium homeostasis and altered contractile properties of MHS skeletal muscle. PMID- 3028487 TI - Alternations in lipid membrane fluidity and the physical state of cell-surface sialic acid in zinc-deficient rat erythrocyte ghosts. AB - Erythrocyte ghosts, prepared from the blood of rats fed zinc-deficient diets, were evaluated for membrane fluidity and surface sialic acid properties using spin-labeled probes and electron spin resonance (ESR) spectroscopy. These physical parameters of the erythrocyte ghosts from the zinc-deficient group were compared to those for erythrocyte ghosts obtained from ad libitum and pair fed controls consuming zinc-adequate diets. As the animals became progressively zinc deficient, the erythrocyte ghost membranes became more fluid than those from the control groups. In addition, the apparent rotational correlation time of Tempamine spin probes on surface sialic acid residues was smaller for the zinc deficient group, indicative of an increased rotational mobility of the spin label. These results suggest that zinc deficiency can have pronounced effects on the physical state of membrane bilayer lipids and cell surface carbohydrates and supports the view that many of the pathological signs of zinc deficiency are due to a general membrane defect. PMID- 3028488 TI - Detergent inhibition of nitric-oxide reductase activity. AB - Gas chromatography revealed that exposure of extracts of the denitrifiers 'Achromobacter cycloclastes', Paracoccus denitrificans, Pseudomonas aeruginosa and Pseudomonas perfectomarina to Triton X-100 inhibited reduction of NO to N2O, and thus concomitantly inhibited reduction of NO2- to N2O. After exposure of extracts to Triton X-100, the ratio of H+ consumed to NO2- added decreased from approx. 2.0 (for untreated extracts) to approx. 1.5, which indicated that NO2- was reduced to NO by the treated extracts. Addition of a CHAPS-soluble extract (devoid of nitrite reductase activity but rich in nitric-oxide reductase activity) to the Triton X-100-treated extract of P. denitrificans restored capacity for reduction of NO2- on to N2O. Exposure to either the NO that accumulated from reduction of NO2- or to enthetic NO transiently inhibited rates of NO2- reduction in Triton X-100-treated extracts. Use of an Oxides of Nitrogen analyzer indicated that only 5-33% of NO2- reduced by untreated extracts appeared in the stripping gas as NO, whereas 80-95% of NO2- reduced by Triton X-100 treated extracts was recovered as NO. PMID- 3028486 TI - Spin-labeled amphotericin B: synthesis, characterization, biological and spectroscopic properties. AB - A biologically active spin-labeled derivative of amphotericin B has been synthesized by the nucleophilic addition of amphotericin B to 4-(2-iodoacetamido) 2,2',6,6'-tetramethylpiperadine-N-oxyl in dimethyl-sulphoxide at 40 degrees C. The derivative is a moderately water-soluble compound which displays the same biological activity of the parental compound against the sensitive organism Leishmania mexicana; also, the rates of proton-cation exchange induced by the two compounds in large unilamellar liposomes are indistinguishable. The ESR spectra of spin-labeled amphotericin B in lipid vesicles indicate a high degree of motion, very similar to that encountered for the compound in aqueous solutions at neutral pH and in deoxycholate micelles, and suggest that the structures formed by the antibiotic in membranes are composed by a small number of molecules. In contrast, the spectra of the labeled antibiotic in ethanol, diethyl ether and dimethylformamide indicate restricted motion and exchange interactions, probably resulting from the micellar aggregation induced in these media. Ascorbate at 10 mM is able to reduce completely the nitroxide group of the labeled antibiotic in lipid vesicles in less than 30 s, indicating that an asymmetric disposition of the antibiotic molecules across the membrane is capable of inducing its biological and ionophoric properties. Ni2+ and Cu2+ produce moderate exchange broadening of the ESR signal of spin-labeled amphotericin B in lipid vesicles; the comparison of this phenomenom with the exchange broadening produced by the same ions in the ESR spectrum of 2,2',6,6'-tetramethylpiperidine-N-oxyl in water solution suggests an specific Cu2+-amphotericin B interaction in membranes. PMID- 3028490 TI - Electron spin resonance studies of the interaction of oxidoreductases with 2,6 dimethoxy-p-quinone and semiquinone. AB - Previous electron spin resonance studies have demonstrated that the decay of ascorbyl plus semiquinone radicals, produced in an aqueous mixture of ascorbate and 2,6-dimethoxy-p-quinone, is accelerated by ascites cells. This effect was concluded to involve a sulfhydryl-containing NAD(P)H-enzyme, and work on cultured cell lines showed that on neoplastic transformation the activity against the radicals was increased. We show here that at least three disulfide oxidoreductases are able to quench the radicals in a similar way to that of viable cells. Glutathione reductase (EC 1.6.4.2) in the presence of NADPH and oxidised glutathione, and dihydrolipoamide dehydrogenase (EC 1.8.1.4) with NADH and lipoamide, are found to accelerate the radical decay by reducing the quinone or semiquinone. DT-diaphorase (EC 1.6.99.2) in the presence of NAD(P)H can also achieve this by reducing the quinone directly. Lipoamide dehydrogenase and glutathione reductase are also capable of reducing nitroxide spin labels, a finding considered of relevance to the reported reduction of such spin labels by neuroblastoma cells. PMID- 3028491 TI - Regulation of intracellular pH in LLC-PK1 cells studied using 31P-NMR spectroscopy. AB - 31P-NMR spectroscopy was used to monitor intracellular pH (pHi) in a suspension of LLC-PK1 cells, a renal epithelial cell line. The regulation of intracellular pH (pHi) was studied during intracellular acidification with 20% CO2 or intracellular alkalinization with 30 mM NH4Cl. The steady-state pHi in bicarbonate-containing Ringer's solution (pHo 7.40) was 7.14 +/- 0.04 and in bicarbonate-free Ringer's solution (pHo 7.40) 7.24 +/- 0.04. When pHo was altered in nominally HCO3(-)-free Ringer's, the intracellular pHi changed to only a small extent between pHo 6.6 and pHo 7.6; beyond this range pHi was linearly related to pHo. Below pHo 6.6 the cell was capable of maintaining a delta pH of 0.2 pH unit (inside more alkaline), above pH 7.6 a delta pH of 0.4 unit could be generated (inside more acid). During exposure to 20% CO2 in HCO3(-)-free Ringer's solution, pHi dropped initially to 6.9 +/- 0.05, the rate of realkalinisation was found to be 0.071 pH unit X min-1. After removal of CO2 the pHi increased by 0.65 and the rate of reacidification was 0.056 pH unit X min-1. Exposure to 30 mM NH4Cl caused a raise of pHi by 0.48 pH unit and an initial rate of re-acidification of 0.063 pH unit X min-1, after removal of NH4Cl the pHi fell by 0.58 pH unit below the steady-state pHi, followed by a subsequent re-alkalinization of 0.083 pH unit X min-1. Under both experimental conditions, the pHi recovery after an intracellular acidification, introduced by exposure to 20% CO2 and by removal of NH4+, was found to be inhibited by 53% and 63%, respectively, in the absence of sodium and 60% and 72%, respectively, by 1 mM amiloride. These studies indicate that 31P-NMR can be used to monitor steady-state intracellular pH as well a pHi transients in suspensions of epithelial cells. The results support the view that LLC-PK1 cells use an Na+-H+ exchange system to readjust their internal pH after acid loading of the cell. PMID- 3028489 TI - Chemical modification of guanidinium groups of vasoactive intestinal peptide. AB - Molecular characterization of receptors depends on the availability of ligand derivatives carrying a reactive group to covalently link the active sites. Two vasoactive intestinal peptide (VIP) derivatives, each labeled either at the two arginine residues 12 and 14 or singly in position 14, were prepared. In the first case, this was achieved by a selective chemical modification using azidophenylglyoxal. In the second, the amino acids of VIP, buried in the active site of the receptor, were protected and one arginine residue of bound VIP was successfully modified using azidophenylglyoxal. The two molecules were resolved by radioimmunocompetition and reversed phase high performance liquid chromatography. Identification of sites of labeling was achieved by tryptic peptide mapping and amino acid analysis. One derivative (Az-Bz-Arg14-VIP) retains a high binding affinity for the receptor and was found to be biologically active. The present method yields a derivative which is useful in structural analysis of the receptor. PMID- 3028492 TI - Assessment of the role of Ca2+ mobilization from intracellular pool(s), using dantrolene, in the glycogenolytic action of alpha-adrenergic stimulation in perfused rat liver. AB - To identify the role of Ca2+ mobilization from intracellular pool(s) in the action of alpha-adrenergic agonist, the effects of dantrolene on phenylephrine induced glycogenolysis were investigated in perfused rat liver. Dantrolene (5 X 10(-5) M) inhibited both glycogenolysis and 45Ca efflux induced by 5 X 10(-7) M phenylephrine. The inhibition by dantrolene was observed in the presence and absence of perfusate calcium. In contrast, dantrolene did not inhibit glycogenolysis induced by glucagon. To confirm the specificity of dantrolene action on calcium release in liver, experiments were also carried out using isolated hepatocytes. Dantrolene did not affect phenylephrine-induced production of inositol 1,4,5-trisphosphate. The compound did inhibit a rise in cytoplasmic Ca2+ concentration induced by phenylephrine both in the presence and absence of extracellular Ca2+. Thus, these results suggest that calcium release from an intracellular pool is essential for the initiation of alpha-adrenergic stimulation of glycogenolysis in the perfused rat liver. PMID- 3028493 TI - Glucagon treatment of rats activates the respiratory chain of liver mitochondria at more than one site. AB - The rate of reduction of ferricyanide in the presence and absence of antimycin and ubiquinone-1 was measured using liver mitochondria from control and glucagon treated rats. Glucagon treatment was shown to increase electron flow from both NADH and succinate to ubiquinone, and from ubiquinone to cytochrome c. 3-(3,4 dichlorophenyl)-1,1-dimethylurea (DCMU) was shown to inhibit the oxidation of glutamate + malate to a much greater extent than that of succinate or duroquinol. Spectral and kinetic studies confirmed that electron flow between NADH and ubiquinone was the primary site of action but that the interaction of the ubiquinone pool with complex 3 was also affected. The effects of various respiratory chain inhibitors on the rate of uncoupled oxidation of succinate and glutamate + malate by control and glucagon treated mitochondria were studied. The stimulation of respiration seen in the mitochondria from glucagon treated rats was maintained or increased as respiration was progressively inhibited with DCMU, 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone (DBMIB), 2-heptyl-4 hydroxyquinoline-n-oxide (HQNO) and colletotrichin, but greatly reduced when inhibition was produced with malonate or antimycin. These data were also shown to support the conclusion that glucagon treatment may cause some stimulation of electron flow through NADH dehydrogenase, succinate dehydrogenase and through the bc1 complex, probably at the point of interaction of the complexes with the ubiquinone pool. The effects of glucagon treatment on duroquinol oxidation and the inhibitor titrations could not be mimicked by increasing the matrix volume, nor totally reversed by aging of mitochondria. These are both processes that have been suggested as the means by which glucagon exerts its effects on the respiratory chain (Armston, A.E., Halestrap, A.P. and Scott, R.D., 1982, Biochim. Biophys. Acta 681, 429-439). It is concluded that an additional mechanism for regulating electron flow must exist and a change in lipid peroxidation of the inner mitochondrial membrane is suggested. PMID- 3028494 TI - Serotonin-induced alterations in inositol phospholipid metabolism in human platelets. AB - When human platelets were incubated for 5 min with [32P]orthophosphate and then stimulated with serotonin, the 32P content of phosphatidylinositol (PI) increased within seconds, compared with the control. The 32P content of phosphatidylinositol 4-phosphate (PIP) and phosphatidylinositol 4,5-bisphosphate (PIP2) only slightly increased during the first minute after addition of serotonin and became more apparent on prolonged stimulation. These changes were not caused by serotonin-induced change in the specific activity of ATP. Using inorganic phosphate determination for the chemical quantification of different inositol phospholipid pools, we found that the platelet PI content remained nearly constant; the amount of PIP increased while that of PIP2 decreased. When the platelets were first prelabeled for 80 min with [32P]orthophosphate, the changes in 32P-labeled inositol phospholipids after addition of serotonin were similar to their changes in mass. When the platelet inositol phospholipids were labeled with myo-[2-3H]inositol, serotonin induced an increase in [3H]inositol phosphates. From these data, it is concluded in addition to the earlier-reported effects on phospholipid metabolism (de Chaffoy de Courcelles, D. et al. (1985) J. Biol. Chem. 260, 7603-7608) that serotonin induces: a very rapid formation of PI; and alterations in inositol phospholipid interconversion that cannot be explained solely as a resynthesis process of PIP2. PMID- 3028495 TI - Comparative study on the stimulation of superoxide production in guinea-pig eosinophils by the calcium ionophore A23187. AB - The effect of calcium and/or magnesium on O2- production by guinea-pig eosinophils stimulated by the calcium ionophore A23187 was studied in comparison to neutrophils. In the absence of calcium, A23187 did not stimulate O2- production in resting eosinophils and neutrophils, regardless of the presence of extracellular magnesium. The A23187-induced O2- production by both cells increased linearly with extracellular Ca2+ concentrations. However, the concentration of Ca2+ required for maximum O2- production in eosinophils was about 10-times lower than that required of neutrophils. The addition of Mg2+ strongly inhibited O2- production, especially in eosinophils at low Ca2+ concentrations. The intracellular Ca2+ concentration was lower in eosinophils than in neutrophils in the resting state, and the enhancement of the intracellular Ca2+ concentration in response to A23187 was much lower in eosinophils than in neutrophils. The activation of the NADPH-dependent O2(-) forming enzyme (NADPH oxidase) in eosinophils depended on extracellular calcium, as observed in O2- production. However, the NADPH oxidase activity in the particulate fraction from A23187-stimulated eosinophils was only slightly affected by the addition of divalent cations or EDTA. The compound W-7 (N-(6 aminohexyl)-5-chloro-1-naphthalenesulfonamide hydrochloride), a calmodulin antagonist, significantly inhibited O2- production by both cells. On the other hand, the compound H-7 (1-(5-isoquinolinesulfonyl)-2-methylpiperazine dihydrochloride), a protein kinase C antagonist, was less effective on O2- production than was W-7. H-7 had little effect on O2- production of eosinophils. These findings suggest that NADPH oxidase might be activated by a smaller Ca2+ concentration through the calmodulin-dependent reaction. This was not observed with protein kinase C, at least in eosinophils. PMID- 3028496 TI - Investigations on the role of Golgi-mediated, ligand-receptor processing in the activation of granulocytes by chemoattractants: differential effects of monensin. AB - Human granulocytes were exposed to different concentrations of the ionophore monensin for 20 min at 37 degrees C. Subsequent exposure to 50 nM of the chemoattractant fMet-Leu-[3H]Phe for up to 30 min at 37 degrees C resulted in a receptor-mediated uptake that was inhibited 80% at a monensin concentration of 30 microM. 50% inhibition was observed at 1-10 microM monensin with no significant change in fMet-Leu-Phe dose dependency. Subcellular fractionation of cells treated with monensin, indicated that the low density UDP-galactosyltransferase activity associated with internalized receptor-fMet-Leu-Phe complexes in untreated cells was absent. The high density galactosyltransferase activity cosedimenting with specific granule markers, however, was unaffected. Monensin also inhibited chemotaxis toward fMet-Leu-Phe as measured by migration of granulocytes through millipore filters and fMet-Leu-Phe induction of polarized morphology. Incubation of cell suspensions with up to 30 microM monensin, both before and during measurement of fMet-Leu-Phe stimulated superoxide production, did not affect the magnitude, kinetics, or transiency of the radical generation. Monensin did, however, shift the dose dependency of superoxide production of fMet Leu-Phe to higher concentrations. These differential effects of monensin suggest that endocytosis of complexes of the chemoattractant and receptor is not involved in the activation or termination of the fMet-Leu-Phe stimulated superoxide production. They also are consistent with a role for receptor modulation and processing in the chemotactic response. PMID- 3028497 TI - Phosphorylation of the alpha-subunit of (Na+ + K+)-ATPase by carbachol in tissue slices and the role of phosphoproteins in stimulus-secretion coupling. AB - This study examined the changes in protein phosphorylation in response to cholinergic (muscarinic) stimulation of salivary secretion in the rat submandibular gland. Carbachol stimulation was associated with phosphorylation in a number of protein bands as detected by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis and autoradiography. The molecular masses (Mr) of two proteins, in which the amount of phosphorylation more than doubled in response to carbachol, were 22,000 and 96,000. The Mr 96,000 protein precipitated at 120,000 X g while most of the Mr 22,000 protein remained in the supernatant at this speed. The effect of carbachol on the phosphorylation of the Mr 22,000 and 96,000 proteins was blocked by atropine, indicating that the cholinergic receptor involved is muscarinic. The time course of phosphorylation of the Mr 22,000 protein consisted of a rapid increase in phosphorylation within the first min of carbachol stimulation. This increased phosphorylation persisted for less than 1 min. The increased phosphorylation of the Mr 96,000 protein also occurred within the first min but it persisted for at least 10 min. However, removal of the muscarinic agonist, carbachol, resulted in the rapid dephosphorylation of this protein. When the plasma membranes were purified, the Mr 96,000 protein was phosphorylated by ATP in the presence of Na+ and Mg2+. It was dephosphorylated by K+. This proves that the Mr 96,000 dalton protein is the alpha-subunit of the (Na+ + K+)-ATPase. PMID- 3028499 TI - [The role of Vavilov-Cherenkov radiation in visual sensations induced by protons]. AB - The sensitivity of human eyes to ionizing particles and the mechanism of their detection were studied. The experimental data obtained by protons with very different intensity of Cherenkov radiation in the vitreous humour (460 and 1850 MeV) showed that Cherenkov radiation plays an essential role in the visual sensation. PMID- 3028498 TI - Differences in arachidonic acid release, metabolism and leukotriene B4 synthesis in human polymorphonuclear leukocytes activated by different stimuli. AB - Products of the 5-lipoxygenase pathway were analyzed after different stimuli in human polymorphonuclear leukocytes prelabeled with 3H-arachidonic acid. Upon stimulation with the Ca2+ ionophore, A23187, polymorphonuclear leukocytes generate 118.2 +/- 18 pg [3H]dihydroxyeicosatetraenoic acids (diHETEs, including 3H-leukotriene B4 and its 6-trans-stereoisomers), after exposure to serum coated zymosan (35.8 +/- 9 pg) and N-fMet-Leu-Phe (39.5 +/- 9 pg). Conversion of 3H arachidonic acid paralleled its release after A23187 and fMet-Leu-Phe exposure leaving only 13.8 +/- 7% and 13.6 +/- 3% of the released 3H-arachidonic acid unmetabolized, respectively. In contrast, after stimulation with serum-coated zymosan only a small fraction of the released 3H-arachidonate was converted to 5 lipoxygenase products leaving 73.0 +/- 5% of the released 3H-arachidonic acid unmetabolized. In parallel, leukotriene B4 synthesis was studied in unlabeled polymorphonuclear leukocytes, resulting in 40 +/- 15 ng upon A23187 stimulation, 4 +/- 0.9 ng upon stimulation with fMet-Leu-Phe and 1.8 +/- 0.9 ng after serum coated zymosan, showing a different ratio of leukotriene B4 to 3H-diHETE for A23187 in contrast to serum-coated zymosan and fMet-Leu-Phe. These results indicate that the coupling between the release of the precursor fatty acid and the metabolism via the 5-lipoxygenase pathway differs greatly between different stimuli. PMID- 3028500 TI - [Iron sources forming nitrosyl complexes in animal tissues]. AB - Treatment of perfused mouse liver with nitric oxide does not change the intensities of ESR signals of iron-sulfur proteins characteristic of this tissue. Proceeding from this evidence and also from the ratio between the iron content in these proteins and dinitrosyl iron complexes (complexes 2.03) formed in the liver when it contacts with NO, it is concluded that iron-sulfur proteins are not involved in the formation of complexes 2.03. It seems that only the loosely bound form of non-heme iron-free iron is involved in this process. PMID- 3028501 TI - [Distribution and changes in properties of ferromagnetic iron particles administered to the animal body]. AB - The registration of ESR signals of organs and tissues in the wide range of temperatures permits to study properties and distribution of ferromagnetic particles in animal organisms. High dispersed powder (HDP) of iron (particle dimension--50-100 nm) was administered subcutaneously to mice in the doses 2 and 100 mg/kg weight of animals. One week after the administration HDP was accumulated in the animal organs under study. Two weeks after the treatment of mice with HDP in the dose of 2 mg/kg the ESR signals with g = 2.1 appeared in the animal tissues: liver, spleen, kidney, heart and lungs. Six weeks after the treatment the ESR signals of the studied tissues did not differ from those in control animals. PMID- 3028502 TI - [Computer simulation of ESR signals of liver paramagnetic centers]. AB - Changes of paramagnetic centres concentration characterized by g-factors values of 1.94, 2.2, and 2.03 in the rat liver were studied by ESR method under acute intoxication by diethylnitrosamine (DENA) and at preliminary threefold treatment of animals with butylhydroxytoluene (BHT). A protective effect of BHT can be explained by its stabilizing action of the membrane structures. A comparison has been carried out with a similar study of paramagnetic centres in the experiment of chronic intoxication by DENA. A simulation was performed of the liver tissue ESR spectra by means of special computer program. The parameters of simulated ESR spectra of the liver tissue with due regard for ESR signal g 2.03 corresponded to the parameters of the experimental spectra. Confirmations were obtained for the nature and number of paramagnetic centres in the liver tissue. PMID- 3028503 TI - [General principles of receptor and neuron function]. AB - Experimental data supporting hypothesis about mechanical part in both neuron and receptor intracellular mechanism are presented. PMID- 3028504 TI - [Determination of the correlation time of the lysozyme molecule and lysozyme inhibitor complex using a spin-label method]. AB - Temperature relationships of rotational correlation times (tau M) of lysozyme molecules were studied using viscosity method based on the model of slow feebly anisotropic rotation of label N-O-group. Protein is mainly associated in 0.01 phosphate buffer (pH 7.3) at 5-30 degrees C. Formation of the complex of lysozyme with competitive inhibitor (3-N-AGA) leads to changes of tau M. It may be due to a decrease of polymerization degree and increase of the protein packing. It is suggested that two-component form of ESR spectra of different spin labeled proteins reflects their common ability for pulsations between more and less compact conformers due to the thermal relative movements of domains and subunits. PMID- 3028505 TI - [Study of molecular mobility in biological membranes by 2-mm band ESR spectroscopy]. AB - Temperature relationship was measured of ESR spectra of spin probes--derivatives of fatty acids whose nitroxyl fragments are incorporated into surface and inner layers of phosphatidylcholine liposomes (T = 180-260 K), as well as of the probe in model ethanol solution (T = 110-220 K). Data on the nature and rotation frequency of probe nitroxyl fragments were obtained from the analysis of the spectra. It was shown that the frequency of the probes movement in the systems under study corresponds to the slow movement region, i.e. it is not above 10(7) s 1. PMID- 3028506 TI - Effects of a missense exonic mutation in cytochrome b gene, observed on isolated mitochondrial complex III of Saccharomyces cerevisiae: consequence for the antimycin binding site. AB - The mitochondrial complex III was isolated from a wild type strain of Saccharomyces cerevisiae PS409 and from two mutants, PS490 and PS493, carrying a missense mutation in the structural gene of cytochrome b (in exons B1 and B4 respectively). These mutants synthesize cytochrome b in variable proportions, but they are unable to grow on a respirable substrate. Strain PS493 does not bind antimycin, whereas strain PS490 contains less cytochromes b and c1 but shows a strong binding to the inhibitor. The complex isolated from the wild type strain or mutant PS493 exhibited a specific cytochrome b and cytochrome c1 heme content of approximately 8 and 4 nmol/mg of protein respectively. This content was about 3 and 2 nmol/mg with PS490, which leads to a molar stoichiometry of 1.3 : 1 for cytochromes b and c1, instead of an 'ideal' ratio of 2 : 1 expected with b-c1 complex, and obtained with the two other strains. This implies that the association (or presence) of b and c1 cytochromes is not pre-requisite for complex III assembly. The wild type complex III isolated from PS409 was found to have a high level of CoQ2H2 activity, using a synthetic coenzyme Q analog as substrate (440 s-1 mol of cytochrome c reduced/mol of cytochrome c1). This activity is fully inhibited by antimycin. The complexes isolated from the two box mutants exhibited no such activity. Analysis of the subunit composition of the three isolated complexes on sodium dodecyl sulfate-gel electrophoresis showed that all the bands belonging to the b-c1 complex were synthesized in both mutants as well as in the wild type strain. Some of them appeared to have slightly diminished, but no specific decrease of a band has been observed in mutant PS493 that does not bind antimycin, with respect to mutant PS490 which binds strongly to the inhibitor. It should be noted that the subunit of about 12-13 kDa, qualified as the antimycin binding protein, is equally present in the three complexes. The results suggest that the loss of antimycin binding in mutant PS493 might be due to conformational perturbations in the modified complex rather than to the disappearance or significant modification of some protein support. PMID- 3028507 TI - Sensitivity of yeast cells to reactive oxygen species generated in the extracellular space. AB - Even when cytoplasmic scavenging activities are plentiful, yeast cells (S. cerevisiae) remain particularly sensitive towards reactive oxygen species generated in the extracellular space (either by the xanthine/xanthine oxidase reaction or by the redox cycling of menadione). A sharp reduction of the extent of cellular alterations when SOD and/or catalase were supplemented in the incubation buffer, points to a contribution of both O-.2 and H2O2 in the toxic process. Although oxygen metabolites as well as t-butylhydroperoxide (tBH), a highly toxic organic peroxide, may be directly responsible for cellular damage, their toxicity is largely reduced in the presence of Desferal. A role of metal ions in potentiating the toxicity points to the involvement of OH. radicals, actually produced in the medium. With tBH, metal cations would be rather active in promoting peroxidative chain reactions. In the case of an extracellular oxidative attack, it may be foreseen that the plasma membrane will form a preferential target. An increased permeability of the plasma membrane towards ionized molecules and uncharged polycarboxylic acids is indeed observed after an oxidative treatment. The loss of selective permeability is, as a rule, correlated with a drop in viability. Early alterations, disrupting the functional organization of the plasma membrane have been sought. The permease involved in the active transport of purine(s) has appeared to be an appropriate marker for checking its functional integrity. This transport function appears to be very sensitive to damage induced by O-.2 generators, particularly under conditions in which the resulting lethality is still kept low and in which the energization of active transport processes remains unimpaired. PMID- 3028508 TI - [Methylation of 3',5'-cGMP phosphodiesterase in retinal photoreceptor membranes]. AB - It was found that the level of methylation of phosphodiesterase (EC 3.1.4.17) and of proteins with Mr 61000 and 21000-26000 Da in isolated preparations of bovine retinal outer segments was sharply increased in the presence of cGMP, but not with cAMP. Preparations of the outer segment phosphodiesterase protein inhibitor, which inhibits the cyclic nucleotide hydrolysis, significantly decreased the methylation level. The degree of the enzyme methylation depended on the functional state of the outer segments: in light-induced preparations it was higher that in the dark ones. PMID- 3028509 TI - [Role of the membranes in lymphocyte interaction with mycoplasmas]. AB - The interaction of Acheleplasma laidlawii cells, derived membranes and liposomes with mouse spleen lymphocytes does not require the energy and participation of electrostatic coupling. The absence of pinocytosis of mycoplasma is demonstrated. Specific membrane receptors don't participate in mycoplasma and lymphocyte binding. The exchange by membrane lipids occurs during the interaction of mycoplasma, membranes and liposomes with lymphocytes; mycoplasma, membranes and liposomes lose saturated fat acids and enrich with cholesterol. On the contrary, lymphocytes lose cholesterol, but absorb fat acids. The intensity of lipid exchange depends on the degree of unsaturation of fat acids in mycoplasma. The carbohydrate transport into lymphocytes is stimulated considerably as a result of interaction of both cells. PMID- 3028511 TI - Failure to alleviate symptoms of schizophrenia with the novel use of an antiviral agent, acyclovir (Zovirax). PMID- 3028510 TI - The HPA axis response to insulin hypoglycemia in depression. AB - Dysfunction of the hypothalamic-pituitary-adrenal (HPA) axis, demonstrated by failure to suppress cortisol secretion after dexamethasone, is found in approximately 50% of patients with major depression (MD). In this study, we examined the response of adrenocorticotrophic hormone (ACTH) and cortisol to insulin-induced hypoglycemia in 20 healthy controls and 18 inpatients with MD [12 dexamethasone suppressors (S) and 5 dexamethasone nonsuppressors (NS)]. After the administration of 0.15 U/kg of regular insulin, both controls and patients with MD showed an increase in plasma ACTH and cortisol levels. Controls had a significantly higher ACTH peak (p less than 0.01) and ACTH increment (p less than 0.01) than MD patients. There were no statistically significant differences between patients who were S and NS. Although baseline plasma cortisol levels were significantly higher in MD patients, there were no significant differences in the peak cortisol or increment in plasma cortisol after hypoglycemia between patients with MD and controls or between patients who were S and those who were NS. These findings suggest that a defect exists in the regulation of the HPA axis at the pituitary level in MD and that this defect is not necessarily reflected in the dexamethasone suppression status of the patient. PMID- 3028512 TI - Serial assessment of corticotropin-releasing hormone response after dexamethasone in depression. Implications for pathophysiology of DST nonsuppression. PMID- 3028513 TI - Platelet [3H]imipramine binding in psychiatric disorders. AB - The Bmax and Kd values for [3H]imipramine binding were measured in platelets from drug-free normal controls and schizophrenic and depressive patients. No differences among groups were found. Exacerbated and remitted patients with either schizophrenia or depression did not differ in platelet [3H]imipramine binding parameters. No correlations were observed between platelet [3H]imipramine binding parameters and measures of symptom severity among actively ill patients with either schizophrenia or depression. PMID- 3028514 TI - Changes in [3H]5-HT uptake and [3H]imipramine binding in platelets after chlorimipramine in healthy volunteers. Comparison with maprotiline and amineptine. AB - In the platelets of normal healthy volunteers (n = 8) taking chlorimipramine (50 mg/day) for 1 week, the saturable uptake of [3H]5-hydroxytryptamine (5-HT) was fully inhibited at the end of the week, but returned to control values after 2 weeks washout. The Bmax of [3H]imipramine binding was decreased by 63% at the end of the treatment and remained significantly decreased below control values after 1 week washout, whereas the Kd values were increased at the end of the treatment, but had returned to baseline values after 1 week washout. The time course of recovery following the administration of chlorimipramine showed some variation between subjects, but it was necessary to wait up to 4 weeks of washout before the Bmax of [3H]imipramine returned to baseline levels. In contrast, neither 1 week treatment with maprotiline (50 mg/day) nor with amineptine (100 mg/day) changed the parameters of [3H]5-HT uptake or [3H]imipramine binding in platelets from healthy volunteers. These results support the following conclusions. (1) [3H]Imipramine binding in platelets can be down-regulated by relatively low, subtherapeutic doses of chlorimipramine. (2) It is possible to dissociate [3H]imipramine binding parameters from [3H]5-HT uptake because the time course of recovery was clearly different, indicating that [3H]imipramine labels a site linked with, but different from, the 5-HT recognition site in the transporter complex. (3) A washout of antidepressants of 4 weeks may be needed when studying the parameters of [3H]imipramine binding in platelets from depressed patients if the previous medication involved chlorimipramine. For antidepressants like maprotiline or amineptine, that act through mechanisms other than inhibition of 5 HT uptake, the time of washout appears to be less critical, although it is not possible to rule out the existence of some secondary modifications influencing the 5-HT transporter complex. PMID- 3028515 TI - Purification of human sperm by a discontinuous Percoll density gradient with an innercolumn. AB - Human sperm were highly purified through the use of a discontinuous Percoll density gradient placed in an inner column of a centrifuge tube. Six ml of 80% Percoll solution were poured into a centrifuge tube with an inner column containing successive 1.0-ml layers of 70, 60, and 40% Percoll solutions. Diluted semen was placed on top of the gradient, and the tube was centrifuged at 600 X g for 30 min using a swing-out rotor. After centrifugation, the majority of the progressive motile sperm were isolated in the sediment; they had a mean motility of 93 +/- 4.1% (n = 10). Other cellular components, including bacteria, remaining in the inner column. The level of bacterial contamination in the purified sperm fraction was below detection for most of the species quantified. The purified sperm were found to be more than 92 +/- 3.2% viable, as judged by dye exclusion, and abnormal sperm were reduced to 5.2 +/- 1.4%. Because of the use of the inner column, the contamination by seminal plasma was negligible in the purified sperm, as estimated by residual protein, fructose, and acid phosphatase activity. PMID- 3028516 TI - Endogenous inhibitors of cyclic adenosine 3',5'-monophosphate-phosphodiesterase in rat epididymis. AB - Stabilization of cyclic adenosine 3',5'-monophosphate (cAMP)-phosphodiesterase (PDE) in 50% glycerol made possible the removal of endogenous inhibitors from tissue extracts by dialysis and the storage of the extracts at -20 degrees C without loss of PDE activity. Dialysates of heat-inactivated epididymal extracts were fractionated by liquid chromatography, and 4 fractions-F2, F5, F7, and F12 were found to contain endogenous inhibitors of PDE. The masses of the fractions required to inhibit low-Km PDE activity by ca. 50% in 430-microliter incubation mixtures were F2, 89 micrograms; F5, 23 micrograms; F7, 275 micrograms; and F12, 1.2 mg. The mechanisms of inhibition of low-Km PDE by the endogenous inhibitors were investigated by kinetic analysis of enzyme-inhibitor interaction. F2 and F12 inhibited PDE competitively; F5 and F7 decreased both apparent Km and Vmax, suggesting an uncompetitive mechanism of inhibition. The high potency of F5 in low concentration suggests that it may be a physiological modulator of low-Km cAMP-PDE activity. PMID- 3028517 TI - Oocyte maturation is inhibited by adenosine in the presence of follicle stimulating hormone. AB - Considerable evidence implicates cyclic 3', 5' adenosine monophosphate (AMP) in the maintenance of meiotic arrest of mammalian oocytes. Since this laboratory previously found that adenosine augmented follicle-stimulating hormone (FSH) stimulated accumulation of cyclic AMP in oocyte-cumulus-complexes (OCC), in the present studies we investigated the possibility that adenosine inhibits maturation of oocytes. In rat OCC cultured in the presence of FSH, adenosine markedly inhibited oocyte maturation in a dose-dependent and biphasic manner. Maximum inhibition of oocyte maturation was seen with 1-30 microM adenosine in the presence of FSH, and half-maximal inhibition occurred with less than 0.3 microM adenosine. High levels of adenosine (100 microM) did not inhibit oocyte maturation in the presence of FSH. In the absence of FSH, adenosine showed little effect on oocyte maturation in the present studies, but increased the maximum inhibition of oocyte maturation produced by FSH approximately twofold. Like adenosine, adenosine triphosphate (ATP), adenosine diphosphate (ADP), and adenosine 5'-monophosphate (AMP) also inhibited oocyte maturation; whereas adenine, guanosine, inosine, and hypoxanthine were inactive at equivalent levels. The metabolism-resistant adenosine analog (2-chloroadenosine) was as active an inhibitor as adenosine. Inhibition produced by the adenine nucleotides may have been direct or due to conversion to adenosine by extracellular nucleotidases. The concentration dependence and purine specificity for inhibition of oocyte maturation are characteristic of an adenosine receptor-mediated process, but direct evidence for such a mechanism was not shown. The effective concentration of adenosine for inhibition of oocyte maturation is within the range of reported levels of adenosine in biological tissues and fluids.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3028518 TI - Gonadotropin-releasing hormone (GnRH) receptor dynamics and gonadotrope responsiveness during and after continuous GnRH stimulation. AB - Gonadotrope responsiveness, serum and tissue levels of luteinizing hormone (LH), and tissue concentration of gonadotropin-releasing hormone (GnRH) receptors in ovariectomized rats were determined during and after continuous GnRH stimulation. Intraperitoneal placement of GnRH-containing osmotic minipumps for 96 h established a rate of GnRH delivery (1 microgram/h) that resulted in stable serum levels of GnRH (500-700 pg/ml). Secretion of LH increased 8-fold within 6 h; however, serum LH returned to pretreatment levels by 24 h, even with continued GnRH stimulation. Tissue concentration of LH was depressed within 48 h of initiation of treatment but levels were restored by 96 h. Tissue levels of GnRH receptor remained elevated during the first 6 h of treatment but were reduced by 60% within 24 h and remained depressed for the duration of treatment. Gonadotrope responsiveness 48 h and 96 h after initiation of treatment was reduced by 50% and 90%, respectively. Removal of the GnRH delivery vehicle resulted in rapid disappearance of GnRH from serum. Dramatic reduction (75%) in circulating levels of LH, and a 2-fold increase in tissue levels of LH and in GnRH receptor concentration were noted within 6 h of minipump removal. Although tissue concentration of GnRH receptor returned to pretreatment levels within 48 h of minipump removal, both basal LH secretion and gonadotrope responsiveness remained depressed even 96 h after cessation of continuous GnRH stimulation. These data indicate that GnRH can "down regulate" its receptor, gonadotrope responsiveness is not obligatorily linked to receptor concentration, and desensitization that follows hyperstimulation represents effects directed at post-receptor loci. PMID- 3028519 TI - Changes in the hypothalamic-hypophyseal axis of mares associated with seasonal reproductive recrudescence. AB - Four groups of mares, representing anestrus (AN; n = 8), early transition (ET; n = 7), late transition (LT; n = 8) and estrus (EST; n = 12) were used to examine changes in the hypothalamus and anterior pituitary during the period of transition from winter anestrus into the breeding season. Mares were of mixed breeding, between the ages of 3 and 20 years, and had shown normal patterns of estrous behavior and ovulation during the breeding season previous to this experiment. Hypothalamic content of gonadotropin-releasing hormone (GnRH) and anterior pituitary content of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) were determined by radioimmunoassay. The number of receptors for GnRH in anterior pituitary tissue was also determined. There was no effect of stage of transition into the breeding season on receptors for GnRH or content of FSH (p greater than 0.05). Likewise, content of GnRH in the hypothalamus did not differ between the four groups (p greater than 0.05). However, pituitary content of LH increased progressively from anestrus to the breeding season (p less than 0.05). Means for the AN, ET, LT and EST groups were 1.1 +/- 0.2, 2.2 +/- 0.3, 6.3 +/- 1.4 and 15.2 +/- 1.8 micrograms LH/mg pituitary, respectively. In addition, serum concentrations of LH associated with the first ovulation of the year for 5 of the EST mares were significantly lower (p less than 0.01) than those associated with the second ovulation of the year.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3028520 TI - Particulate aluminum oxide as a bone graft material. AB - The performance of particulate aluminum oxide (Al2O3) as a bone graft material was evaluated by comparison to two other graft materials, autogenous bone and particulate hydroxylapatite (HA). Holes were drilled through the medial cortex of both the distal femora and proximal tibiae in three dogs. The Al2O3, HA and autogenous bone were packed into the created defects and the rate and extent of defect healing examined. Particulate Al2O3 and HA were also placed on opposite sides of the median sacral crest in a spinal fusion procedure. One dog was sacrificed at 4, 8 and 12 weeks post-operative. The extent and rate of healing was superior with the autogenous bone, while the performance of the HA was superior to that of the Al2O3 at 4 and 8 weeks. However by 12 weeks, the performance of the two non-autogenous materials was equivalent. At 12 weeks the majority of the cancellous defect was infiltrated with viable bone with all three materials. In the spinal fusions, results were not as encouraging, with less bone present within the graft materials as compared to the cancellous defect implantations. PMID- 3028521 TI - Dynamics of superhelical DNA studied by photon correlation spectroscopy. AB - We have conducted photon correlation spectroscopy (PCS) studies on the plasmid pUC8 (2717 bp) in order to elucidate the internal dynamics of this superhelical DNA. We confirm that the first-order autocorrelation function of the scattered light from pUC8 solutions can be separated into two distinct exponential decay components, as first shown by Lewis et al. (R. Lewis, J.H. Huang and P. Pecora, Macromolecules 18 (1985) 944). A thorough analysis of the dependence on scattering vector K of the rates and amplitudes of the two components enables us to assign the slowly relaxing part to the center-of-mass diffusion of the DNA, while the faster component corresponds to rotational, bending and twisting motions of the superhelix. For larger K values the internal motions can be formally expressed in terms of an 'internal diffusion coefficient' Di, whose value of 2.0-2.5 X 10(-11) m2 s-1 is approximately equal to the translational diffusion coefficient predicted for a stiff DNA piece of the persistence length, 65 nm. Comparison of our measured Di values to those predicted from a recent theory of circular worm-like coils (K. Soda, Macromolecules 17 (1984) 2365) shows that the internal motions are faster than the theoretical values. One of the reasons for this discrepancy could be that the theory does not take into account torsional motions, which contribute significantly to the internal dynamics (J.C. Thomas, S.A. Allison, C.J. Appelof and J.M. Schurr, Biophys. Chem. 12 (1980) 177). At low K values, the fast relaxation of superhelical pUC8 is no longer proportional to K2, but reaches a constant value as K approaches zero. This behavior, not seen for the linearized DNA, can be interpreted in terms of rotational diffusion of a flexible rod-like molecule (T. Maeda and S. Fujime, Macromolecules 17 (1984) 2381) and supports an interwound rod-like structure for pUC8 DNA with an average end-to-end distance of 220 nm. PMID- 3028522 TI - Interaction of thymic peptide thymosin alpha 1 with vasoactive intestinal peptide (VIP) receptors. AB - Thymic peptide thymosin alpha 1 (10(-9) to 3 X 10(-7) M) is shown to inhibit the specific binding of [125I]VIP to rat blood mononuclear cells and liver plasma membranes. Thymosin alpha 1 was 160 and 6250 times less potent that VIP at inhibiting [125I]VIP binding to blood mononuclear cells and liver plasma membranes, respectively. Thymosin alpha 1 (10(-10) to 10(-7) M) was weak in stimulating adenylate cyclase activity. Its efficacy is about 25% and 27% that of native VIP in blood mononuclear cells and liver plasma membranes, respectively. Thymosin alpha 1 may behave as a partial VIP agonist in rat. PMID- 3028523 TI - The structure of a cDNA clone corresponding to mouse cardiac muscle actin mRNA. AB - A cDNA library was constructed from mouse cardiac muscle mRNA, and a clone corresponding to part of the mRNA for the cardiac muscle isoform of actin was isolated from this library. The nucleotide sequence of the cloned insert was determined and was found to contain almost the complete amino acid coding region for actin (only codons for the first two amino acids, absent from the mature protein, were lacking) and a substantial portion derived from the 3' untranslated region of the mRNA. Comparison of the latter with the corresponding region in cardiac actin mRNA from man and rat showed that this 3' untranslated region has been subject to conservational pressure during evolution. However a comparison with the corresponding region in skeletal muscle actin mRNAs indicated that the pattern of conservation is quite different in the two striated muscle actin isoforms. PMID- 3028524 TI - Phospholipids as a possible instrument for translocation of nascent proteins across biological membranes. AB - The interaction of phospholipids with precursor proteins, particularly with the mitochondrial precursor protein apocytochrome c is reviewed and integrated with other aspects of protein insertion and translocation, leading to a model for (apo)cytochrome c import into mitochondria, in which phospholipids play a dominant role. PMID- 3028525 TI - Evidence that fasting can induce a selective loss of uncoupling protein from brown adipose tissue mitochondria of mice. AB - The effects of fasting and refeeding on the concentration of uncoupling protein in brown adipose tissue mitochondria have been investigated in mice. Fasting mice for 48 h led to a large decrease in the total cytochrome oxidase activity of the interscapular brown fat pad. Mitochondrial GDP binding and the specific mitochondrial concentration of uncoupling protein also fell on fasting. After 24 h refeeding both GDP binding and the mitochondrial concentration of uncoupling protein were normalized, but there was no alteration in the total tissue cytochrome oxidase activity. Fasting appears to induce a selective loss of uncoupling protein from brown adipose tissue mitochondria, which is rapidly reversible on refeeding. PMID- 3028526 TI - Homologous IRCM-serine protease 1 from pituitary, heart atrium and ventricle: a common pro-hormone maturation enzyme? AB - IRCM-Serine Protease 1 (IRCM-SP1) has recently been isolated and characterized from porcine pituitary anterior and neurointermediate lobes (Cromlish et al., 1986a, J. Biol. Chem. 261:10850-10858; Cromlish et al., 1986b, J. Biol. Chem. 261:10859-10870). This pituitary serine protease was shown to selectively cleave human pro-opiomelanocortin (POMC)-derived peptides at both pairs of basic residues and C-terminal to specific Arg residues, all known to be cleaved in vivo. Here, a similar enzyme was isolated from rat heart atria and ventricles. Rat IRCM-SP1 was shown to be highly specific for the same cleavage sites in POMC, as the porcine pituitary homologue. Furthermore, the rat and the porcine enzymes cleave rat pro-Atrial Natriuretic Factor (pro-ANF 1-126) to yield ANF 103-126, 102-126 and 99-126 in that order of preference. This suggests that in vitro the cleavage sites preferred in pro-ANF resemble those found in brain and hypothalamus. The enzyme is nine times more abundant in atria versus ventricles/mg protein. It is concluded that IRCM-SP1, could well represent a common pro-hormone maturation enzyme for POMC and Pro-ANF and possibly many other pro-hormones. PMID- 3028527 TI - [Action of vasopressin on the electrical and contractile reactions of vascular smooth muscles in animals of various ages]. AB - Vasopressin action on isolated strips of the femoral artery was studied in mature and old rats. Vasopressin has been shown to depolarize smooth muscle cells (SMC) membranes, causing the tonic contraction of the strips. SMC sensitivity to vasopressin in old animals was increased, electrical and contractile reactions were also enhanced. Peculiar tachyphylaxia in response to vasopressin has been found in SMC of mature animals. In old animals SMC tachyphylaxia was far less expressed than in mature ones. Two types of vasopressin receptors (V1-exciting and V2-inhibitory) in vascular SMC were suggested. Vasopressin does not influence cAMP content of the femoral artery in mature and old animals. Specific features of SMC reactions to vasopressin in old animals may underlie the mechanisms of cardiovascular pathology in old age. PMID- 3028528 TI - [Experimental diabetes in mice infected with Coxsackie viruses]. AB - The influence of Coxsackie B4 and AI3 viruses on the pancreas of mice (resistant and susceptible to diabetes) was studied. Glucose intolerance and changes in the synthesis of immunoreactive insulin were detected in all the treated groups of animals. Biochemical changes were more prominent in male DBA/2 mice, infected with Coxsackie B4 virus, in FI (CBA X C57Bl/6) hybrids and in female DBA/2 mice infected with Coxsackie AI3 virus and alloxan. PMID- 3028529 TI - [Protective properties of microcrystalline cellulose in experimental diabetes mellitus in rats]. AB - The experiments on 45 inbred male and female rats with alloxan diabetes have shown that microcrystalline cellulose at a dose of 3 g/day received per os beginning from day 15 to day 42 after alloxan administration stabilizes the animals' body mass, reduces the level of glycemia and glycosylated hemoglobin. A parallel reduction of triglyceride and total cholesterol levels and increase in plasma phospholipid content, compared to the control, are observed, as well as a considerable hypocoagulation shift in the blood aggregation system. PMID- 3028530 TI - [Changes in the ligand affinity of benzodiazepine receptors in the presence of 4,5,6,7-tetrahydroisoxazolo(5,4-c)pyridin-3-ol (THIP)]. AB - 4,5,6,7-tetrahydroisoxazolo-(5,4-c-)pyridin-3-ol (THIP) in concentrations of 100 500 microM inhibited 3H-flunitrazepam (3H FNZ) binding to intact membranes at 0 degrees C in 25 mM TRIS-HCl buffer. The inhibition of 3H FNZ binding was due to the decrease in the affinity and Bmax of 3H FNZ binding. The affinity for benzodiazepines (e.g. BZ-receptor agonists) was reduced in the presence of THIP, whereas the affinity for BZ-receptor antagonists was unchanged. The value of THIP induced shift was dependent on the molarity of TRIS-HCl buffer and the concentration of THIP. The results obtained suggest that THIP-induced shifts have a predictive value in the determination of pharmacological efficacy of BZ receptor ligands. PMID- 3028531 TI - [Role of cyclic 3',5'-adenosine monophosphate in the development of Salmonella infection in an experiment]. AB - The changes in cAMP levels in spleen macrophages of mice infected with low virulent and virulent Salmonella strains and the effect of propranolol on Salmonella reproduction in the spleen, and the outcome of Salmonella-induced infection have been studied. A persistent increase in cAMP levels in spleen macrophages during Salmonella infection caused by virulent Salmonella strains has been demonstrated. Low-virulent Salmonella strains failed to cause the elevation of cAMP levels in spleen macrophages. Propranolol injection to mice prevented intensive Salmonella reproduction in the spleen and diminished the animal mortality rate. PMID- 3028532 TI - [Antibodies interacting with the proteins of the mammary cancer virus (MMTV) in the sera of healthy women]. AB - Using enzyme-linked immunoassay (ELISA), sera of 405 women without breast cancer were screened for the presence of antibodies to mouse mammary tumour virus proteins. 16 donors with positive sera were identified (3.96 +/- 0.97%). An attempt was made to establish the correlation between the age, ovarian Activity, benign breast disease and other factors and the appearance of antibodies to mouse mammary tumour virus. No statistically significant differences have been revealed among women aged 20-29 (3.92%), 30-39 (4.69%), 40-49 (4.08%) and 50-59 (2.38%), as well as in premenopausal (4.31%) and postmenopausal (4.5%) women and women with simple fibromatous disease (4.76%) and without any mammary gland pathology. PMID- 3028533 TI - [Development of the duodenal glands in rats on a high cellulose diet]. AB - Wistar weaned rats were kept on cellulose-rich diet for 3 months. They revealed the duodenal gland area of increased length, as compared to the control group kept on standard pellets. Glandular-duodenal index, that is a relative indication of gland development, also rose. Cellulose-fed rats showed a lower pH of the stomach contents. The correlation between pH and gland development has been established. Possible reasons for a more intensive gland enlargement in herbivorous animals are being discussed. PMID- 3028534 TI - Diversity of the effect of recombinant tumor necrosis factors alpha and beta on human myelogenous leukemia cell lines. AB - In vitro characteristics of response to recombinant tumor necrosis factors alpha (rTNF-alpha) and beta (rTNF-beta) were studied in six human myelogenous leukemia cell lines. Heterogeneity of the response to rTNF was observed, and one line (K562) was resistant. No enhancement of cell growth was noted in any cell line. The dose-response curves for rTNF-alpha were characteristically sigmoid, the maximum inhibitory effect occurring between 25 and 200 ng/mL. Nonresponsiveness within this range indicated resistance that could not be overcome, even with very high doses of rTNF alpha. A similar response of sensitive and resistant lines occurred after exposure to rTNF-beta. The clonogenic cells were more sensitive than the overall population to the action of rTNF alpha, and prolonged exposure was necessary for manifestation of the effect. Concomitant exposure to recombinant interferon-alpha (rIFN-alpha) increased the response of two cell lines to rTNF-alpha, but no clear synergistic action could be demonstrated. The addition of rIFN-gamma was without effect. Variations in the rTNF-alpha-induced proliferative response could not be explained by differences in the number of binding sites per cell or their affinity for rTNF-alpha. That the clonogenic cells showed a higher sensitivity than the whole population might indicate a preferential effect on more primitive, actively proliferating cells with high growth potential. PMID- 3028535 TI - Activation of neutrophil superoxide production by concanavalin A can occur at low levels of intracellular ionized calcium. AB - The effect of concanavalin A (Con A) on the concentration of ionized intracellular calcium [( Cai++]) in human granulocytes (PMN) was monitored using the fluorescent calcium indicator and chelator, Quin2. The addition of Con A to PMN resulted in a rise in [Cai++] that was markedly enhanced by the presence of Ca++ in the external buffer. The onset of the increment in [Cai++] preceded the onset of O2- production. The rise in [Cai++] induced by Con A is not transient, with a new, higher steady-state level of [Cai++] being attained within five minutes. The addition of alpha-methylmannoside (alpha-MM) one minute after Con A resulted in the return of [Cai++] to the original baseline level and the cessation of O2- production. The addition of a second stimulus (such as arachidonic acid) to these cells resulted in a second increment in [Cai++] and the return of O2- production. Thus the rise in [Cai++] induced by Con A is tightly coupled to the activation, inactivation, and reactivation of the O2- generating system by Con A. Further experiments were undertaken to assess the possible requirement for the rise in [Cai++] in the activation of PMN by Con A. PMN could be depleted of Cai++ by loading with Quin2 in the absence of extracellular Ca++. These Ca++- depleted PMN can be induced to produce O2- after treatment with PMA but not with Con A. The addition of Ca++ to Ca++ -depleted PMN results in a return of [Cai++] to the normal resting level of Ca++ -replete PMN. The time required to return to baseline is a function of the concentration of intracellular Quin2. The addition of Ca++ to Ca++ -depleted, Con A-treated PMN results in the elevation of [Cai++] and the production of O2-. Over a tenfold range of intracellular Quin2 concentration, the onset of O2- production always occurred at [Cai++] that were less than the normal resting level. Thus, activation of the O2- generating system by Con A can occur at [Cai++] which are much lower than the incremental level induced by Con A in Ca++ -replete PMN. Supporting this is the observation that only a very small increment in [Cai++] is induced by Con A in PMN cytoplasts, even though Con A could induce O2- production in the Quin2-loaded cytoplasts.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3028537 TI - Chromosome abnormalities in AIDS-associated lymphadenopathy. AB - Cytogenetic studies were performed on direct and 24-hour culture preparations of eight lymph node biopsies from seven patients with acquired immunodeficiency syndrome (AIDS) or AIDS-related complex (ARC)-associated lymphadenopathy in whom histological evidence of lymphoma was not detected. Three of these seven had chromosomal abnormalities, including chromosome instability in one and clonal chromosomal abnormalities in two; one of the latter was a t(8;14)(q24;q32). The remaining five showed normal karyotypes. Epstein-Barr virus (EBV) titers were elevated in all three patients that exhibited chromosome abnormalities, two of whom later developed malignant lymphoma. A control group of five patients with reactive lymphadenopathy not associated with AIDS failed to reveal chromosomal aberrations, but elevated EBV titers were present in two. These data are consistent with current views on the role of EBV and chromosome change in the development of lymphoma in immunodeficient states and suggest that karyotypically abnormal AIDS-related lymphadenopathy represents a prelymphomatous proliferation. PMID- 3028536 TI - Thrombocytopenia associated with pregnancy in a patient with type IIB von Willebrand's disease. AB - Thrombocytopenia may accompany variant (type IIB) von Willebrand's disease (vWD) and is thought to result from binding of the abnormal von Willebrand factor (vWF) to the patient's platelets with subsequent platelet aggregate formation and clearance. We have studied a patient with type IIB vWD who became thrombocytopenic during two pregnancies. During the third trimester of pregnancy, her platelet counts dropped to 20,000 to 30,000/microL, and an increase in the intermediate-sized vWF multimers was seen on agarose gel electrophoresis. During this time her platelet-rich plasma showed spontaneous platelet aggregation, and her plasma caused spontaneous aggregation of normal washed platelets. Antibody to platelet glycoprotein Ib completely blocked the spontaneous platelet aggregation, while antibody to platelet glycoprotein IIb/IIIa did not block the response at the concentrations used. Inhibitors of platelet function that elevate platelet cyclic AMP also blocked the response, but aspirin had no effect on the spontaneous platelet aggregation. The patient illustrates that the platelet counts in one individual can vary greatly in type IIB vWD and that the thrombocytopenia that occurs can appear under physiologic conditions that stimulate the endogenous production of the patient's abnormal vWF. The mechanisms leading to spontaneous platelet aggregation and thrombocytopenia appear to be similar to those described for other patients with type IIB vWD. PMID- 3028538 TI - Sodium fluoride mimics effects of both agonists and antagonists on intact human platelets by simultaneous modulation of phospholipase C and adenylate cyclase activity. AB - Using intact human platelets, we studied the effect of sodium fluoride (NaF) on platelet aggregation and release reaction and correlated the functional changes to intracellular events specific for either agonist-induced or antagonist-induced platelet responses. At lower concentrations, with a peak activity between 30 and 40 mmol/L, NaF induced aggregation and release of adenosine 5'-triphosphate (ATP) that was associated with increased formation of inositol phosphates, a rise in cytosolic free Ca2+, and phosphorylation of 20-kd and 40-kd proteins. At NaF concentrations greater than 40 mmol/L, aggregation and ATP release decreased dose dependently in parallel with a decrease in Ca2+ mobilization, whereas neither inositol phosphate formation nor 40-kd protein phosphorylation was reduced. At these concentrations, NaF caused a dose-dependent transient rise in platelet cyclic adenosine 3',5'-monophosphate (cAMP) levels that was sufficient to account for the observed reduction in Ca2+ mobilization, aggregation, and ATP release. Stimulated cAMP levels started declining rapidly within 30 seconds of addition of NaF, however. Similarly, prostacyclin (PGI2)-induced cAMP accumulation was temporarily enhanced but subsequently suppressed by NaF, suggesting either stimulation of a cAMP phosphodiesterase or delayed inhibition of adenylate cyclase. Evidence for the latter was provided by the finding that NaF pretreatment of platelets resulted in partial inhibition of PGI2-stimulated cAMP formation in the presence of the cAMP phosphodiesterase inhibitor 3-isobutyl-1 methyl-xanthine (MIX). We conclude that NaF exerts a dual (stimulatory and inhibitory) effect on adenylate cyclase in intact platelets that is accompanied by simultaneous activation of a phosphoinositide-specific phospholipase C; in addition, a cAMP phosphodiesterase may be activated. PMID- 3028539 TI - Cell surface phenotyping of megakaryoblasts. AB - Surface phenotypic characterization of megakaryoblasts, identified by platelet peroxidase activity, was investigated in four patients who showed increased proliferation of megakaryoblasts: one patient with typical features of acute leukemia, one presenting with acute myelofibrosis, and two with Down's syndrome in whom blasts disappeared spontaneously (transient abnormal myelopoiesis, TAM). MY10 and/or MY9 antigens were expressed on the surface of some megakaryoblasts, but MY7, and MY4, antigens specific to granulocytic or monocytic cells, were not. Some megakaryoblasts were positive for only anti-HLA-DR antibodies. It was speculated that, during the differentiation of the megakaryocytic lineage, MY9 antigen appears transiently on the surface of megakaryoblasts that have lost HLA DR antigens and have gained the glycoprotein IIb/IIIa antigen. This study also demonstrated that the proliferating blasts in some patients with TAM were mainly megakaryoblasts and suggested that the target cells in TAM are CFU-GEMM. PMID- 3028540 TI - Normal epithelial antigens recognized by GB3 and GB5 are diminished in intraductal and lost in infiltrating human breast carcinomas. AB - GB3 and GB5 are mouse monoclonal antibodies raised against human amnion. GB3 detects the epidermal basement membrane, and GB5 reacts with the junctional substances between the epithelial cells. These two antibodies were studied on breast tissues by indirect immunofluorescence. All (n = 8) of the normal mammary glandular basement membranes and epithelia reacted strongly with GB3 and GB5. In the intraductal carcinomas (n = 10), although nine out of ten tissues were reactive, their reactivities were greatly reduced. More strikingly, in the infiltrating carcinomas (n = 15), none of the tumor specimens were recognized by GB3 and only one out of fifteen biopsies was moderately reactive with GB5, indicating that the syntheses of the antigens of GB3 and GB5 were discontinued in the metastatic breast adenocarcinomas. These data suggest that these antigens may contribute to the interaction of the epithelium with the extracellular matrix and the intercellular binding between the epithelial cells; the loss of these antigens may facilitate the wide dissemination of tumor cells. PMID- 3028541 TI - Levels of nine potentially toxic elements in Idaho fish manures. PMID- 3028542 TI - Persistence of phorate in different soils with and without amendments and its degradation by a Pseudomonas sp. PMID- 3028543 TI - Bronchioloalveolar carcinoma. PMID- 3028544 TI - Royal Free disease: perplexity continues. PMID- 3028545 TI - Effect of dietary cholesterol on plasma cholesterol concentration in subjects following reduced fat, high fibre diet. AB - One hundred and sixty eight subjects participated in a randomised crossover study to determine whether halving or doubling the present dietary cholesterol intake from eggs had any influence on blood cholesterol concentration in people following current dietary recommendations. During the first eight weeks all participants were advised to follow a reduced fat diet (26% total energy for hyperlipidaemic patients, 35% total energy for normolipidaemic volunteers) with an increased ratio of polyunsaturated to saturated fatty acids. This background diet was continued throughout the 16 week experimental period, during which participants ate either two or seven eggs a week. A small but significant increase in total cholesterol was seen after four weeks in the group eating seven eggs a week compared with that in the group eating two eggs a week, but this was no longer apparent after eight weeks. Previous studies suggesting that dietary cholesterol has a greater effect on the serum cholesterol concentration either have been carried out against a background of a higher fat intake or have contrasted extreme cholesterol intakes. A further reduction in dietary cholesterol seems to be unnecessary in those people who have already reduced their intake of saturated fat and increased the ratio of polyunsaturated to saturated fatty acids and fibre rich carbohydrate. PMID- 3028546 TI - Neurological and neurosurgical approaches in the management of malignant brain tumours. PMID- 3028547 TI - Antiviral chemotherapy. AB - Specific antiviral chemotherapy now has an established role in the treatment of many virus infections, particularly those caused by herpesviruses. This change is one of the major medical developments of the last decade. There is reason to believe that further advances will occur, and we can look forward to a range of antiviral drugs comparable to those available for bacterial infections. PMID- 3028548 TI - Actions of alpha, beta-methylene ATP and 6-hydroxydopamine on sympathetic neurotransmission in the vas deferens of the guinea-pig, rat and mouse: support for cotransmission. AB - alpha-Adrenoceptor antagonists (prazosin or phentolamine) reduced the contractile response to field stimulation of the isolated vasa deferentia of guinea-pig, rat and mouse. alpha, beta-Methylene ATP (alpha, beta-MeATP) reduced that portion of the contraction which was resistant to alpha-adrenoceptor blockade. alpha, beta MeATP (1-800 microM) did not affect action potential conduction in the guinea-pig vas deferens nerves, and (up to 10 microM) did not reduce the stimulation-evoked overflow of [3H]-noradrenaline from this tissue. Spontaneous excitatory junction potentials (s.e.j.ps) in the majority of cells of guinea-pig, rat, and mouse vasa were abolished by alpha, beta-MeATP (0.1-10 microM). In a small number of cells s.e.j.ps were resistant to the actions of alpha, beta-MeATP (10 microM). Excitatory junction potentials (e.j.ps) in the majority of cells in vasa of all species studied were abolished by alpha, beta-MeATP (1-10 microM). E.j.ps elicited in some 'resistant' cells demonstrated marked facilitation characteristics. It is concluded that alpha, beta-MeATP inhibits s.e.j.ps and e.j.ps by a postjunctional action. In all species pretreatment of animals with 6 hydroxydopamine produced a marked reduction in noradrenaline (NA) content (as determined by fluorescence histochemistry) and abolished e.j.ps, findings which suggest that e.j.ps originated from sympathetic nerves. The results support the hypothesis that NA and ATP are co-transmitters in the sympathetic nerves of rodent vasa. PMID- 3028549 TI - Activation and desensitization of presynaptic alpha 2-adrenoceptors after inhibition of neuronal uptake by antidepressant drugs in the rat vas deferens. AB - The isolated field-stimulated vas deferens of the rat (0.1 Hz, 3 ms, 30-40 V) was used to study the relationship between the in vivo inhibition of neuronal uptake of noradrenaline (NA) by cyclic antidepressant drugs and the subsequent activation/desensitization of presynaptic alpha 2-adrenoceptors. Receptor activation was indirectly measured by quantifying the ability of each drug to inhibit basal twitch responses after their acute administration. Receptor desensitization was also indirectly measured by quantifying the ability of the drugs to reduce the inhibitory effects of selective alpha 2-adrenoceptor agonists on the electrically-induced twitch responses after their long-term administration. The acute in vivo administration of desipramine and other antidepressants (0.5-10 mg kg-1; i.p.; 2 h) resulted in dose-dependent inhibitions of the basal twitch responses which were rapidly reversed to control values by idazoxan (10-5 M). In vitro, desipramine and other antidepressants also inhibited in a concentration-dependent manner (10(-9)-10(-5) M) the twitch responses. In rats pretreated 12 h earlier with reserpine (1 mg kg-1; i.p.) or oxypertine (4 mg kg-1; i.p.), desipramine (10 mg kg-1; 2 h) did not induce inhibition of the basal twitch responses or it induced a smaller effect, respectively. For the various antidepressants the degree of inhibition of the basal twitch responses (desipramine greater than protriptyline greater than nortriptyline greater than maprotiline = imipramine greater than amitriptyline greater than viloxazine greater than iprindole much greater than zimelidine) was highly correlated (r = 0.914) with the potency for blockade of [3H]-NA uptake into rat brain synaptosomes. Clonidine and xylazine inhibited in a concentration dependent manner (10(-9)-10(-6) M) the twitch responses. The long-term (7-14 days) administration of antidepressants or cocaine (10 mg kg-1, i.p.) resulted in significant decreases in sensitivity to clonidine or xylazine. Short-term (3 days) treatment with desipramine did not reduce the sensitivity to clonidine. The results indicate that the acute in vivo inhibition of NA neuronal uptake by antidepressants leads to the activation (through endogenous NA) of presynaptic inhibitory alpha 2-adrenoceptors which results in inhibition of the twitch responses. In contrast, prolonged in vivo inhibition of NA reuptake is followed by a slow desensitization process of the same receptors which results in a reduction of sensitivity to clonidine. PMID- 3028550 TI - The effects of N-(cyclopropylmethyl)-19-isopentylnororvinol (M320), a potent agonist at kappa- and mu-opiate receptors, on urine excretion of rats. AB - The effects of N-(cyclopropylmethyl)-19-isopentylnororvinol hydrochloride (M320) on urine excretion by rats were investigated. Further studies, using rabbit isolated vas deferens, investigated its interactions with kappa-opiate receptors. The output of urine for a 2 h period after M320, administered subcutaneously to normally hydrated Long Evans rats, showed a bell-shaped dose-response relationship, the maximum effect occurring after 10 micrograms kg-1. Urinary retention contributed to but did not fully account for the weaker diuresis after high doses. Attenuation of the ascending portion of the dose-response curve to M320 occurred after 1 and 10 mg kg-1 but not 0.1 mg kg-1 of naltrexone intraperitoneally. M320 in low doses (3-10 micrograms kg-1) caused a small but significant increase in sodium excretion. M320 (30 micrograms kg-1) reduced both sodium and potassium excretion. M320 (10 micrograms kg-1 s.c.) did not increase the volume of urine voided in 2 h by Brattleboro rats showing diabetes insipidus, even when urine excretion was reduced to normal by 1 week of vasopressin replacement. The volume of urine voided in 4 h by Brattleboro rats was progressively reduced to zero by M320 (10-100 micrograms kg-1 s.c.). Urinary retention contributed to but did not account for this reduction. Plasma levels of immunoreactive arginine vasopressin (ir-AVP) were reduced in both normal and dehydrated Long Evans rats after doses greater than 1 microgram kg-1 M320 s.c. In vitro, M320 caused persistent inhibition of twitches of the electrically stimulated rabbit vas deferens (IC50 1.7 nM). 8 These data suggest that M320 has potent opioid agonist activity at K-receptors and at higher concentrations stimulates mu- receptors. In the rat, its activity on K-receptors is associated with diuresis and suppression of plasma vasopressin levels. The antidiuresis seen after high doses may be due to its activity on mu-receptors, possibly at a central site. PMID- 3028551 TI - Presynaptic muscarinic and alpha-adrenoceptor-mediated regulation of GABA release from myenteric neurones of the guinea-pig small intestine. AB - The effects of cholinomimetic and sympathomimetic drugs on the release of [3H] gamma-aminobutyric acid ([3H]-GABA) evoked by high K+ from the isolated small intestine of the guinea-pig were investigated, in the presence of tetrodotoxin. Acetylcholine and oxotremorine, at concentrations ranging from 10(-9) to 10(-6) M inhibited the evoked release of [3H]-GABA in a concentration-dependent manner, while nicotine was without effect. Scopolamine and pirenzepine inhibited the effect of oxotremorine, while hexamethonium had no effect. The IC50 values for scopolamine and pirenzepine of the oxotremorine (3 X 10(-8) M)-induced inhibition were 1.02 X 10(-9) M and 9.78 X 10(-10) M, respectively. Noradrenaline, but not isoprenaline inhibited the evoked release of [3H]-GABA. Clonidine (10(-10)-10(-6) M) reduced the evoked release of [3H]-GABA in a concentration-dependent manner, but phenylephrine had no effect. The inhibitory effect of clonidine was antagonized by yohimbine but not by prazosin. These findings provide evidence for the localization of M1-muscarinic and alpha 2-adrenoceptors on GABAergic nerve terminals and their involvement in the presynaptic control of the release of GABA from the guinea-pig small intestine. PMID- 3028552 TI - Electrophysiology of neuroeffector transmission in the isolated, innervated trachea of the guinea-pig. AB - Intracellular recordings were made from cells of the guinea-pig trachealis muscle. Some cells were electrically quiescent while others exhibited spontaneous slow waves. In quiescent cells, stimulation of the cervical vagus nerve evoked transient depolarization. Occasionally there was a single depolarization, but more often there were several fluctuations in potential. In spontaneously active cells, vagal stimulation induced a transient increase in amplitude of the slow waves without affecting their frequency. Depolarizing responses could be obtained with a single pulse applied to the vagus nerve, and responses increased in amplitude with number of pulses (up to 16 pulses), and with frequency of stimulation (up to 20 Hz). Depolarization did not give rise to spike discharge. Responses to vagal stimulation were blocked by atropine. In the presence of neostigmine, vagally-mediated depolarization was augmented and abortive spikes were observed in a number of cells. In quiescent cells, repetitive stimulation of the sympathetic stellate ganglion evoked slight hyperpolarization. In spontaneously active cells, sympathetic stimulation evoked attenuation, or temporary cessation of slow wave discharge, with or without hyperpolarization. Sympathetic-induced hyperpolarization and suppression of slow waves were both blocked by propranolol, but unaffected by phentolamine. Electrical changes associated with sympathetic stimulation may be of minor importance in the initiation of relaxation. PMID- 3028553 TI - Abnormal vascular phosphoinositide hydrolysis in the spontaneously hypertensive rat. AB - The production of [3H]-inositol phosphates was studied in labelled segments of aorta from spontaneously hypertensive rats (SHR) and Wistar Kyoto (WKY) controls at 5 and 19 weeks, either unstimulated or in the presence of noradrenaline. Basal hydrolysis of inositol phospholipids was significantly enhanced in young SHR (P less than 0.05) compared to controls but this difference was no longer detected at 19 weeks. Noradrenaline increased [3H]-inositol phosphate accumulation in both SHR and WKY, but maximal hydrolysis was significantly greater in WKY (P less than 0.01), although the ED50 was similar in both groups of animals. These data demonstrate that phosphatidylinositide hydrolysis is enhanced in the young hypertensive rat at the time blood pressure is rising, but that this activity has declined by the time hypertension has reached an established phase. In addition, alpha 1-agonist induction of inositol phospholipid hydrolysis differs in the two species of animals, being reduced in genetically mature hypertensive rats. PMID- 3028554 TI - Influence of the vascular endothelium on agonist-induced contractions and relaxations in rat aorta. AB - The influence of the vascular endothelium on agonist-induced contractions and relaxations has been measured using intact segments of rat aorta. Contiguous rubbed segments were used as controls. Angiotensin II, histamine, noradrenaline, U46619 and UK14304 contracted both rubbed and intact tissues. The threshold spasmogenic concentrations of these agonists were lower in rubbed tissues than in intact preparations. The sensitivity and responsiveness of tissues to angiotensin II, histamine, noradrenaline and UK14304 were greater in rubbed than in intact tissues. Acetylcholine and histamine relaxed the established spasms of intact tissues but not those of rubbed preparations, These relaxant effects of acetylcholine were abolished by pre-incubation with haemoglobin. In the presence of prazosin, noradrenaline or UK14304 relaxed established contractions in intact tissues. These effects were antagonized by idazoxan or by pre-incubation with haemoglobin. In intact preparations, idazoxan had no effect on the spasmogenic sensitivity and responsiveness to UK14304. Pre-incubation with haemoglobin augmented the spasmogenic actions of noradrenaline, U46619 or UK14304 in intact tissues, but had no effect on these responses in rubbed preparations. Tissue concentrations of cyclic GMP were greater in intact than in rubbed tissues. A concentration of acetylcholine (10 microM) evoking just maximal mechanical inhibition produced a significant increase in cyclic GMP concentration in intact preparations. However, no detectable changes in cyclic GMP concentration were produced by UK14304 (10 microM) or by acetylcholine (30 nM), concentrations which were equi-effective in inhibiting mechanical activity. In the presence of threshold spasmogenic concentrations of noradrenaline, the contractile effects of angiotensin II were augmented and became comparable to those observed in rubbed preparations. In the presence of greater concentrations of noradrenaline, angiotensin II always produced an additional contraction. It is concluded that the presence of the vascular endothelium limits the spasmogenic action of a variety of agonists. Although spasmogens like noradrenaline and UK14304 can stimulate the release of endothelium-derived relaxing factor (EDRF) via alpha 2 adrenoceptors, the inhibitory effects of EDRF largely result from the spontaneous release of this substance. PMID- 3028555 TI - Stimulation of receptors of gamma-aminobutyric acid modulates the release of cholecystokinin-like immunoreactivity from slices of rat neostriatum. AB - Slices of rat dorsal neostriatum were incubated in Krebs-Henseleit medium and the release of cholecystokinin-like immunoreactivity (CCK-IR) was induced by veratridine or high concentrations of K+. It was investigated whether drugs which act at receptors for gamma-aminobutyric acid (GABA) affected the release. The GABAA-receptor agonists muscimol and isoguvacine enhanced the veratridine-induced release of CCK-IR. This effect was abolished by the GABAA-receptor antagonist, bicuculline. When used alone, bicuculline decreased the release. The GABAB receptor agonist, (-)-baclofen, decreased the veratridine-induced release of CCK IR. The stereoisomer (+)-baclofen, which has low intrinsic activity, had no effect when used alone, but antagonized the effect of (-)-baclofen as did delta amino-n-valeric acid, another antagonist at GABAB-receptors. When the release of CCK-IR was stimulated by K+ (40 mM) in the presence of tetrodotoxin, it was no longer affected by GABAA-receptor agonists or antagonists. Thus, their sites of action were probably not in the immediate vicinity of the nerve-endings which release CCK-IR. Under these conditions, stimulation of GABAB-receptors still reduced the release of CCK-IR. Therefore, it is concluded that these receptors are in the immediate vicinity of or even on the terminals which release CCK-IR. PMID- 3028556 TI - Nerve pathways involved in adrenergic regulation of electrical and mechanical activities in the chicken rectum. AB - Peripheral nerve pathways responsible for adrenergic inhibition of mechanical and electrical activities in the chicken rectum and receptors mediating the adrenergic inhibition were investigated in isolated extrinsically-innervated rectum of the chicken. Electrical stimulation of the anal end (Ra) or the ileal cut end (Ri) of Remak's nerve, or perivascular nerves (P) elicited relaxation of the rectum pretreated with atropine (0.5 microM) and hexamethonium (0.3 mM) to block the cholinergic and non-cholinergic, non-adrenergic excitatory innervations. Ri stimulation was much less effective than Ra and P stimulations. The relaxation was shown to be related to cessation of spontaneous spike discharge of the longitudinal muscle which was accompanied by membrane hyperpolarization. The inhibitory effects elicited by Ra and P stimulations, which were prolonged beyond the period of the stimulation, were converted to transient ones by propranolol (3.4 microM). Phentolamine (2.6 microM) reduced effectively the residual effects. In contrast, the effects of Ri stimulation were little affected by these drugs. The present results provide evidence for the existence of two nerve pathways responsible for direct adrenergic inhibitory innervation to the chicken rectum, one running orally in Remak's nerve trunk, leaving it and descending in the branches to the rectum, and the other running as the perivascular nerves along the arterial supplies of the rectum. The direct innervation is mediated predominantly by beta-adrenoceptors. PMID- 3028558 TI - Differential effects of Ca antagonists on the noradrenaline release and contraction evoked by nerve stimulation in the presence of 4-aminopyridine. AB - We previously reported that verapamil, nicardipine and diltiazem inhibited both neurotransmitter release and contraction evoked by transmural nerve stimulation (TNS) in the canine saphenous vein. To evaluate whether the three Ca antagonists act on the nerve endings by inhibiting Ca2+ influx, the effects of the three antagonists were studied in the presence of 4-aminopyridine (4-AP) 3 X 10(-4) M on the TNS-evoked tritium overflow and contraction of canine saphenous veins preloaded with [3H]-noradrenaline. 4-AP increased both tritium overflow and contraction evoked by TNS, but did not enhance the contraction induced by exogenous noradrenaline (10 nmol). In the veins pretreated with 4-AP, verapamil (3 X 10(-5) M) and nicardipine (10(-5) M and 3 X 10(-5) M) caused no significant effects on the TNS-evoked tritium overflow, but they still inhibited the contraction. Diltiazem (10(-5) M and 3 X 10(-5) M) significantly inhibited both responses to TNS in the veins pretreated with 4-AP, the effects being nearly equipotent to those in the absence of 4-AP. The (-)-cis isomer of diltiazem (10( 5) M and 3 X 10(-5) M), which is about 100 times less potent than diltiazem in inhibiting Ca2+ influx, inhibited both responses to TNS in the presence of 4-AP to almost the same degree as diltiazem. When 4-AP was added after the Ca antagonists, it reversed the TNS-evoked tritium overflow inhibiting actions of verapamil (3 X 10(-5) M) and nicardipine (3 X 10(-5) M) much more effectively than that of diltiazem (3 X 10(-5) M). Tetracaine (4 X 10(-6) M) significantly inhibited the TNS-evoked tritium overflow and contraction, which were unaffected by 4-AP. Sodium salicylate (10(-2) M) failed to modify the inhibition of TNS evoked tritium overflow following diltiazem (3 X 10(-5) M), but it enhanced that of tetracaine (4 X 10(-6) M). Verapamil but not diltiazem and nicardipine significantly increased the spontaneous tritium overflow from veins pretreated with 4-AP. The present study together with previous results suggests that diltiazem but not verapamil and nicardipine may inhibit the TNS-evoked neurotransmitter release through an action other than inhibition of Ca2+ influx into the adrenergic nerve endings, allowing an inhibition of the resulting contraction. PMID- 3028557 TI - A 7-phenyl substituted triazolopyridazine has inverse agonist activity at the benzodiazepine receptor site. AB - To investigate further the structural requirements for benzodiazepine (BZD) receptor ligands, we synthesized SR 95195, [7-phenyl-3-methyl-1,2,4-triazolo-(4,3 b) pyridazine], a positional isomer of the 6-phenyl-triazolo-pyridazines, which were the first non-BZD derivatives to exhibit high affinity for the BZD receptor and BZD-like activity in vivo. In vitro, SR 95195 displaced specifically bound [3H]-flunitrazepam from rat cerebellar and hippocampal membranes with respective IC50 values of 4 and 8 microM. In vivo, SR 95195 lacked BZD-like activity. At high doses SR 95195 induced clonic seizures in mice (threshold convulsant dose: 150 mg kg-1; CD50: 160 mg kg-1 i.p.) which were antagonized by Ro 15-1788. At non convulsant doses (25 mg kg-1 i.p. and 100 mg kg-1 i.p.) SR 95195 significantly decreased punished responding in an operant conflict procedure in the rat, suggesting SR 95195 has intrinsic anxiogenic activity. SR 95195, in mice, reversed the anticonvulsant and myorelaxant actions of diazepam 3 mg kg-1, orally (respective ED50 values: 45 mg kg-1 i.p. and 44 mg kg-1 i.p.). In an operant conflict test in rats, SR 95195 at non-anxiogenic doses, antagonized the disinhibitory action of diazepam 4 mg kg-1, i.p. (ED50: 8.6 mg kg-1, i.p.), but not that of pentobarbitone 15 mg kg-1, i.p. It is concluded that SR 95195 has the pharmacological profile of an inverse BZD agonist and that displacing the phenyl from the 6- to the 7-position in the triazolopyridazine series causes a shift from agonist to inverse agonist type activity at the BZD receptor site. PMID- 3028559 TI - Effects of tachykinins on inositol phospholipid hydrolysis in slices of hamster urinary bladder. AB - Tachykinin-stimulated inositol phospholipid hydrolysis was examined in slices of hamster urinary bladder. In the presence of lithium, to inhibit inositol monophosphatase activity, substance P, eledoisin and related tachykinins induced large, dose-dependent increases in [3H]-inositol monophosphate accumulation. The responses to substance P and eledoisin were not antagonized by the cholinoceptor antagonist, atropine. The rank order of potency for various tachykinins was kassinin greater than neurokinin A greater than neurokinin B greater than eledoisin greater than physaelamin greater than substance P greater than substance P methyl ester. The synthetic analogue [p-Glu6, D-Pro9]SP (6-11) was considerably more potent than its L-prolyl stereoisomer at stimulating inositol phospholipid hydrolysis. These results suggest that in the hamster urinary bladder, tachykinin-induced inositol phospholipid breakdown is mediated via tachykinin receptors of the SP-E type, as opposed to the SP-P type. PMID- 3028562 TI - Sodium bicarbonate abuse: a case report. AB - Sodium bicarbonate abuse is unusual, and rarely reported. A patient was extensively investigated at several hospitals for a recurrent hypokalaemic metabolic alkalosis; it transpired that she had been abusing sodium bicarbonate for 8 years and had gained hospital admission at will by taking large amounts. She also showed features of the Munchausen syndrome. PMID- 3028560 TI - A facilitatory effect of bicuculline on the enteric neurones in the guinea-pig isolated colon. AB - Changes in the efficiency of the peristaltic reflex, acetylcholine (ACh) output and motor responses to transmural and periarterial nerve stimulation produced by bicuculline and gamma-aminobutyric acid (GABA) receptor desensitization were investigated in the guinea-pig isolated colon. Bicuculline, at concentrations unable to affect spontaneous colonic motility and lacking anticholinesterase activity, produced a dose-dependent increase of both the efficiency of the peristaltic reflex and the stimulated ACh output. Such effects could not be observed in GABA-desensitized preparations. A frequency-dependent potentiation of the cholinergic excitatory and non-adrenergic non-cholinergic (NANC) inhibitory responses to transmural stimulation was also observed in the presence of bicuculline. Conversely bicuculline exhibited an inhibitory effect on the relaxation induced by periarterial nerve stimulation. Acute GABA-desensitization was unable to affect the contractile responses to transmural stimulation, the ACh output and the efficiency of the peristaltic reflex. On the contrary, desensitization was able to mimic the effects of bicuculline on the inhibitory responses to both transmural and periarterial nerve stimulation. Our results are consistent with a significant role played by an intrinsic GABAergic pathway in the modulation of both cholinergic excitatory and NANC inhibitory neurones. The hypothesis is advanced that a feed-back modulation carried out through bicuculline-sensitive GABAergic synapses could operate during the propagation of peristaltic motor activity. PMID- 3028563 TI - Parvovirus infection in hospital practice. AB - Eleven cases of human parvovirus (HPV) infection were diagnosed on the basis of positive IgM serology over a 9-month period in two London hospitals. These cases accounted for almost 30% of viral illness in which HPV serology had been requested. Ten of the cases presented with joint symptoms and/or rash and one case presented with evidence of a severe autoimmune haemolytic anaemia. Six patients had lymphadenopathy. Clinical and laboratory features of the cases are presented. HPV DNA was sought but not found in synovial fluid from one patient. PMID- 3028565 TI - Local GABAergic modulation of noradrenaline release in the rat olfactory bulb measured on superfused slices. AB - Ascending noradrenergic projections from locus coeruleus reaching the rat olfactory bulb (OB) synapse onto gamma-aminobutyric acid (GABA)-ergic interneurons. We tested the hypothesis that these interneurons could locally modulate noradrenaline (NA) release. On superfused OB slices, GABA (10, 100 and 1000 microM) enhanced potassium-evoked release of newly accumulated [3H]NA. This raises the possibility that GABAergic interneurons could establish a functional link between olfactory deutoneuron activity and NA release in the rat OB. PMID- 3028564 TI - Changes in cortical beta-adrenergic receptor density and neuronal sensitivity to norepinephrine accompany morphine dependence and withdrawal. AB - Radioligand binding experiments were carried out in conjunction with electrophysiological recordings in vivo in the parietal cortex in rats to assess changes in postsynaptic beta-adrenergic receptor function that result after chronic administration of morphine and during morphine withdrawal. Chronic treatment of rats with morphine for 14 days resulted in a 38% increase in the density of beta-adrenergic receptors in the parietal cortex, as measured by the binding of the specific antagonist [3H]dihydroalprenolol (DHA). In comparison, following withdrawal in the chronic morphine-treated animals, the number of specific [3H]DHA binding sites in this same cortical region was decreased 25%, when compared to saline-treated controls. These alterations in cortical beta adrenergic receptor density were not accompanied by a significant change in the dissociation constant (Kd) for [3H]DHA or in the inhibitory constants (Ki) for the specific agonists norepinephrine and isoproterenol. Microiontophoretic testing revealed that the changes in beta-adrenergic receptor density found in parietal cortex after chronic morphine treatment and during morphine withdrawal were accompanied by a selective increase and decrease, respectively, in the sensitivity of cerebrocortical neurons in the same region to beta-adrenergic stimulation. These results suggest that changes in central adrenergic function might be related to the formation and/or expression of dependence on morphine. PMID- 3028561 TI - [3H]-verapamil binding to rat cardiac sarcolemmal membrane fragments; an effect of ischaemia. AB - The [3H]-verapamil binding activity of rat cardiac sarcolemmal fragments was studied, using membranes harvested from non-perfused, aerobically-perfused and ischaemic hearts. Glass-fibre filters were found to contain specific, high affinity--(KD 38 +/- 3.1 nM) [3H]-verapamil binding sites--making them unsuitable for use in [3H]-verapamil binding studies. Incubation of membranes from non perfused hearts in a medium containing 150 mM NaCl, 1 mM CaCl2 and 50 mM Tris revealed two populations of [3H]-verapamil binding sites. When centrifugation instead of filtration was used to separate bound and free [3H]-verapamil, high affinity sites with a KD of 0.57 +/- 0.19 microM and a Bmax of 38 +/- 5.2 pmol mg 1 protein, and low affinity sites with a KD of 78 +/- 27.5 microM and a Bmax of 2.9 +/- 1.3 nmol mg-1 protein were detected. However, only low affinity binding sites could be detected in membranes which had been incubated in a cation-free medium containing 50 mM Tris. [3H]-verapamil binding to the low and high affinity sites was saturable, reversible, stereospecific and displaceable by D600 greater than diltiazem greater than Ca2+ but not by nifedipine, nitrendipine, nisoldipine or prazosin. The two populations of binding sites survived aerobic perfusion and 60 min ischaemia at 37 degrees C. Ischaemia reduced the Bmax and KD but selectivity was maintained. PMID- 3028566 TI - A GABAergic depolarizing potential in the hippocampus disclosed by the convulsant 4-aminopyridine. AB - Intracellular recordings from hippocampal pyramidal cells in the CA1 subfield of the 'in vitro' slice in the presence of 4-aminopyridine (4-AP, 5-50 microM) revealed a long-lasting (up to 1.5 s) depolarizing potential which occurred either spontaneously or following orthodromic stimulation. This potential was: capable of blocking both direct and synaptic activation of the cell; sensitive to bath application of low concentrations of bicuculline methiodide; and associated to an extracellular current sink in the dendrites as suggested by the extracellular field potentials recorded at different levels along an axis perpendicular to the stratum pyramidale. It is concluded that the long-lasting depolarizing potential evoked by 4-AP is caused by the activation of GABA receptors localized in the dendritic region of the CA1 subfield. PMID- 3028567 TI - Blockade of cholinergic receptors by an irreversible antagonist, propylbenzilylcholine mustard (PrBCM), in the rat cerebral cortex causes deficits in passive avoidance learning. AB - Studies were made on the effects of blockade of muscarinic acetylcholine (mACh) receptors in the rat cerebral cortex on learning and memory assessed by performance of a step-through passive avoidance task. Bilateral injection of propylbenzilylcholine mustard (PrBCM) into both the frontal and parietal cortex at doses of 2.25 X 4 to 22.5 X 4 micrograms decreased mACh receptors dose dependently, as assessed by [3H]quinuclidinyl benzilate binding studies. When the training trial of a step-through passive avoidance task was performed 24 h after injection of 7.5 X 4 to 22.5 X 4 micrograms PrBCM into the frontal and parietal cortex, and then a retention test was made 24 h after the training trial, the treated rats showed shorter latencies than controls. In contrast, injection of PrBCM into the occipital cortex had no significant effect on performance in the test. These results confirm the notion that cholinergic neurotransmission in the cerebral cortex, especially the frontoparietal cortex, is important in learning and memory. The effects of injection of PrBCM (22.5 X 4 micrograms) into the frontoparietal cortex on 3 postulated phases of the learning and memory process (i.e. registration, retention and recall) were also examined. When PrBCM was injected 24 h before the training trial, no retention of the task was observed 14 days after the training trial. However, when PrBCM was injected 24 h after the training trial, retention of the task 14 days after the training trial was not affected. When PrBCM was injected 3-24 h after the initial training trial, the latencies in the retention test examined 24 h later were shorter than those of control rats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3028568 TI - The development of alpha 2-adrenoceptors in the rat kidney: correlation with noradrenergic innervation. AB - The development of the renal alpha 2-adrenoceptors in the rat was studied by binding renal membranes of prenatal and postnatal rats with p-[3,5 3H]aminoclonidine (PAC), a selective alpha 2-adrenoceptor agonist. The results demonstrate that prior to birth and on the day of birth [3H]PAC binding is present and reflects a single linear binding affinity (Kd approximately equal to 1.0 nM). In contrast, data from postnatal day 3 demonstrate the addition of a second, lower affinity binding site (Kd approximately equal to 7.0 nM) that develops rapidly during the ensuing weeks of life. Both high and low affinity alpha 2-adrenoceptor binding sites continue to increase in receptor density through 30 weeks of age; however, the binding affinity of each site remains relatively stable. The sympathetic noradrenergic innervation of the rat kidney is present prenatally and rapidly develops during the first 3 weeks of life with peak noradrenaline concentrations being reached at approximately prenatally and rapidly develops during the first 3 weeks of life with peak noradrenaline concentrations being reached at approximately postnatal day 21. Immunohistochemical staining demonstrates that tyrosine hydroxylase-positive axons rapidly develop in the kidney during the same period, indicating that the increase in noradrenaline is related to expansion of renal sympathetic innervation. PMID- 3028569 TI - Neuronal birthdate underlies the development of striatal compartments. AB - The striatum of the mammalian forebrain is composed of two complementary functional compartments, the patches and the matrix. By injecting [3H]thymidine at different embryonic times and sacrificing the rats as young adults, we found that the earliest neurons to leave the mitotic cycle were restricted to the patch compartment. Neurons that became postmitotic at later times preferentially joined the matrix compartment. Distinctive periods of cell proliferation may underlie pattern formation throughout the developing forebrain. PMID- 3028570 TI - Morphologic features of embryonic neocortex grafts in adult rats following frontal cortical ablation. AB - Embryonic cortex from 19-day fetuses was transplanted in a medial frontal cortex wound cavity of 105-day-old male rats. Nissl-stained tissue revealed little internal laminar organization. Graft sections impregnated by the Loyez method exhibited bands of myelinated fibers surrounding implants as well as long interconnecting and swirl-like fiber fascicles within the implant. Tissue processed histochemically for acetylcholinesterase and choline acetyltransferase revealed enzyme-positive fibers and cell bodies within the grafts. Cytochrome oxidase histochemistry revealed regional variations in the metabolic activity of the grafts. In summary, although our frontal cortex grafts exhibit many of the morphological features seen in intact frontal cortex, the organization of these components within the implant is dissimilar to normal cortical tissue. PMID- 3028571 TI - Hirano bodies and other cytoskeletal abnormalities develop in fetal rat CNS grafts isolated for long periods in peripheral nerve. AB - Dissociated cells from the telencephalic region of 12-day-old rat embryos were cultured in vitro for 3-5 days and transplanted into segments of the sciatic nerve of adult inbred rats. Transplanted progenitor cells survived, differentiated, and expressed various morphological and molecular features characteristic of neurons and glia. Six to twelve months after grafting, many neurons underwent changes compatible with an alteration of their cytoskeleton. These included: (1) a strong perikaryal immunoreactivity to the monoclonal antibody RT97, directed against the 200-kDa phosphorylated neurofilament subunit and (2) the formation of Hirano bodies within dendrites. Similar cytoskeletal abnormalities are seen as part of the spectrum of changes that occur in some human neurodegenerative diseases and in aging. The approach we have used may provide new possibilities for the study of the pathogenesis of these lesions under controlled laboratory conditions. PMID- 3028572 TI - Acute cold-restraint stress affects alpha 2-adrenoceptors in specific brain regions of the rat. AB - The effect of acute cold-restraint stress on binding of [3H]rauwolscine to alpha 2-adrenoceptors was investigated in 10 regions of rat brain. Acute stress: significantly decreased the density but had no significant effect on the affinity of binding sites in the hippocampus; decreased density and increased affinity in the amygdala; and increased density and decreased the affinity in the midbrain. Seven other brain regions showed no significant changes in binding parameters in response to stress. These results, together with previous findings in this laboratory showed that alteration by restraint stress of noradrenaline levels in amygdala and hippocampus, but not other regions, indicate an association between neurotransmitter turnover and receptor function. PMID- 3028573 TI - Field potential evidence for long-term potentiation of feed-forward inhibition in the rat dentate gyrus. AB - Trains of high-frequency stimulation to the perforant path cause (i) long-term potentiation (LTP) of the population excitatory post-synaptic potential (EPSP), (ii) a lasting increase in the population spike, and (iii) a lasting alteration of the relationship between the EPSP and population spike (E-S relationship), consisting of a decreased x-intercept and decreased slope of the linear regression. To compare the thresholds of these changes, we applied a series of trains, increasing in duration from below LTP threshold. The EPSP potentiated with about the same low threshold as the reduction in E-S slope, whereas the reduction in E-S x-intercept required longer trains. In the second experiment, LTP of the EPSP was reduced by concurrent high-frequency stimulation of the commissural input and a lasting reduction of the population spike height was observed. In a third experiment, picrotoxin, an antagonist of gamma-aminobutyric acid (GABA)-mediated inhibition, blocked the decrease in slope of the E-S relationship which normally accompanies LTP. These results imply that perforant path/granule cell LTP is normally accompanied by long-term potentiation of a feed forward inhibitory pathway which may involve interneurones. PMID- 3028574 TI - Neurophysiological alterations in caudate neurons in aged cats. AB - These neurophysiological studies provide information on the alterations in functional capacity of neurons in the aging caudate nucleus (Cd) of the cat. The major finding is that there is a marked loss of excitation in the Cd during the aging process. This loss is most apparent in animals 11-14 years of age but is demonstrable in animals 6-7 years of age. Extracellular recording techniques were used to test the ability of Cd neurons to respond to activation of two of their major inputs, the precruciate cortex (CX) and the substantia nigra (SN). Types of responses that were evoked in both 1-3- and 11-14-year groups were similar and consisted of excitation, excitation followed by inhibition of action potentials or inhibition alone without preceding excitation. The frequency of occurrence of these responses was altered in the aged animals when either input was stimulated. In 1-3-year-old cats CX stimulation evoked initially excitatory responses in 75% of the cells tested while in 11-14-year-old cats excitatory responses occurred in 62% of the cells. When the SN was stimulated the decrease in initial excitation was greater (69% in 1-3- vs 35% in 11-14-year groups). In all aged animals but not in 1-3-year-old cats stimulation thresholds were higher (39-79%) for evoking excitatory responses than for evoking inhibitory responses. In order to assess synaptic security, the ability of Cd neurons to respond to iterative stimulation was determined. Distributions of the minimum interval necessary to evoke two excitatory responses were constructed. There was a marked increase in the proportion of longer intervals in the aged animals indicating that the synaptic response was less secure. There was a tendency for more of the responses in aged animals to have shorter latencies. This result was probably due to loss of less secure longer latency responses that are mediated via multisynaptic pathways. These findings indicate that there are functional changes in a population of Cd neurons in aged cats that impair their ability to process information. PMID- 3028575 TI - Opioid receptors in rat neostriatum: radioautographic distribution at the electron microscopic level. AB - The distribution of mu-opioid receptors, selectively labeled in vitro with a monoiodinated Met-enkephalin analog [( 125I]FK 33-824), was analyzed by light and electron microscopic radioautography in sections from the neostriatum of the rat. In the light microscope, patches of high receptor densities were detected amidst a moderately labeled matrix. The number of silver grains, as counted in 1-micron thick plastic-embedded sections, was 3 times greater inside the patches than in the intervening matrix. In both compartments, the proportion of labeled binding sites associated with the neuropil was significantly higher (greater than 70%) than that associated with nerve cell bodies or myelinated fascicles. Quantitative analyses of electron microscopic radioautographs revealed that the majority of silver grains corresponding to specifically bound [125I]FK molecules originated from radioactive sources associated with apposed neuronal membranes. Of the total number of specific binding sites, 53% was associated with axodendritic, 18% with axoaxonic and 3% with axosomatic interfaces. The occurrence of multiple labeled foci along the plasma membrane of certain perikarya and dendrites suggested that some of the binding sites might be associated with somato/dendritic elements. The high incidence of labeling along axoaxonic interfaces indicated that others were linked to the membrane of axons and/or axon terminals. A major finding of the present study was that only a small proportion of specific FK binding sites (7% of total) was associated with synaptic junctions. Labeled synapses were primarily of the asymmetric type and were found predominantly on dendritic branches and spines. A few were observed on nerve cell bodies. Labeled symmetric synapses were rare and encountered exclusively on dendritic branches. The high frequency with which specifically labeled binding sites were found to be associated with neuronal interfaces involving axonal processes strongly suggests that even if non junctional these binding sites correspond to functional receptors. Whether these receptors are activated by endogenous ligand molecules released by the labeled terminals themselves or from terminals located at a distance from the labeled interfaces remains to be determined. PMID- 3028576 TI - Morphological and physiological properties of caudal medullary expiratory neurons of the cat. AB - The activity of respiratory neurons in the caudal part of the nucleus retroambigualis (NRA) was recorded intracellularly in decerebrated, spontaneously breathing cats. Spinal projections of these neurons were determined by antidromic stimulation at the C3 segment. A high concentration of bulbospinal expiratory (BS E) neurons was identified in the caudal NRA, whereas the inspiratory (I) neurons, not activated antidromically, were also found to be intermingled in the same region. The BS-E neurons had ramp-like depolarizing potentials during expiration, and repolarized rapidly at the onset of phrenic nerve discharge. The I neurons depolarized abruptly in the early I phase, and repolarized gradually thereafter, namely, they were early-I neurons. Intracellular current injections revealed postsynaptic inhibition of the BS-E neurons during inspiration, as evidenced by inhibitory postsynaptic potential reversal. Using the technique of intracellular labeling with horseradish peroxidase, seven well-stained expiratory cells located in the caudal NRA revealed detailed information about axonal morphology: the axon projected rostrally and dorsomedially for the first 2 mm after emerging from the soma, then turned caudally and ventrally along two different courses, and crossed the midline of the medulla almost at the same rostrocaudal level as the soma. No axon collaterals were observed along the length of the stained portion, indicating that the BS-E neurons cannot influence other respiratory neurons in the brainstem. It has been concluded that the NRA expiratory neurons are involved only in spinal action, and that they receive a postsynaptic inhibition during inspiration.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3028577 TI - Responses to GABA by isolated insect neuronal somata: pharmacology and modulation by a benzodiazepine and a barbiturate. AB - Mechanically dissociated neuronal somata from the thoracic ganglia of Locusta migratoria and Schistocerca gregaria were viable in vitro for hours and were current- and voltage-clamped to record the responses evoked by brief pressure applications of gamma-aminobutyric acid (GABA) in the presence of various modulators. The application of GABA and muscimol, but not baclofen, produced a hyperpolarization and concurrent increase in the membrane conductance. The current underlying this response reversed at -65 mV, was evoked in all cells tested and showed outward rectification. In 6 of 74 Locusta neurones but not in the neurones of Schistocerca, GABA and muscimol evoked a biphasic response. The initial, fast phase was indistinguishable from the GABA-evoked current seen in all neurones. The remaining predominant, slow and long-duration component of the response was an inward current over the membrane potential range 0 to -80 mV, increasing with hyperpolarization. The GABAA antagonists bicuculline and pitrazepin were without effect on the fast GABA response while picrotoxin was a potent blocker of both the fast and the slow GABA responses. Flunitrazepam enhanced the amplitude of the fast response by up to 70% without increasing its duration. Sodium pentobarbital enhanced both the amplitude and the duration of the fast GABA response. We conclude that the locust thoracic neuronal GABA receptor/channel complex resembles the vertebrate GABAA receptor in having associated modulatory receptor sites for benzodiazepines and barbiturates, but differs from it in terms of the pharmacology of the GABA receptor itself. PMID- 3028578 TI - Cholinergic neurons in the rabbit retina: immunocytochemical localization, and relationship to GABAergic and cholinesterase-containing neurons. AB - Cholinergic neurons have been identified in the rabbit retina by an immunocytochemical technique for the localization of choline acetyltransferase. The dendritic processes of these cells form two strata in the inner plexiform layer; these strata show almost no overlap with GABAergic strata (identified with glutamate decarboxylase immunocytochemistry) or with acetylcholinesterase (AChE) containing strata (identified with AChE cytochemistry), but the latter two overlap almost exactly. Implications of these data for the possible mechanisms of directional selectivity are discussed. PMID- 3028579 TI - Redistribution of synaptic vesicles during long-term potentiation in the hippocampus. AB - Synaptic vesicles were quantified 20 min after the induction of long-term potentiation (LTP) in the Schaffer-commissural system of the hippocampus. With LTP, significant increases were found in vesicles attached to the active zone membrane, and in the percentage of vesicles adjacent to the active zone. In addition, as others have reported, overall vesicle density was decreased and spine area was increased. These results suggest that an increased probability of vesicle release may contribute to brain LTP. PMID- 3028581 TI - Excessive cholecalciferol in a layers diet: decline in some aspects of reproductive performance and increased bone mineralisation of progeny. AB - Feeding hens a diet containing 5,000 micrograms (200,000 ICU)/kg of cholecalciferol for four 28-d periods had no adverse effect on hen-day egg production or hatchability. Egg weight, shell quality, food consumption and fertility were significantly decreased in hens fed 5,000 micrograms/kg. of cholecalciferol compared with those fed 24 micrograms (960 ICU) cholecalciferol/kg diet. Plasma calcium increased significantly as the concentration of cholecalciferol was increased in the diet. However, no histologically detectable changes in the viscera or changes in the proportion of bone ash were observed with any concentration of the vitamin. Chicks hatched from dams receiving excessive doses of cholecalciferol (5,000 micrograms/kg) and maintained on a rachitogenic diet for 4 weeks had a significantly higher proportion of tibial ash but there was no effect on either body weight or tibial calcium. PMID- 3028580 TI - Tectal transplants into the occipital cortex of the newborn rat. AB - Embryonic day 15 rostral tectum (presumptive superior colliculus) was transplanted into the occipital cortex of newborn rats. One to two months later, the transplants were visualized and injected with either horseradish peroxidase (HRP) or wheat germ agglutinin conjugated with HRP (WGA/HRP). After the appropriate survival time and processing with either diaminobenzidine (DAB) or tetramethylbenzidine tetrahydrochloride (TMB), HRP-labelled pyramidal cells were found in layer V of the host ipsilateral occipital cortex. Thus, the occipitotectal connections are formed between host and graft despite the fact that the fibers must grow in a direction opposite to their normal course to reach the aberrantly positioned tectal graft. PMID- 3028582 TI - [Effect of rabbit thrombomodulin on the inhibition of thrombin by the antithrombin-heparin complex: role of the acid domain of thrombomodulin]. AB - Acidic and non-acidic forms of rabbit thrombomodulin were studied with regard to their effects on the inhibition of thrombin by antithrombin in the presence of exogenous heparin. The non acidic form was obtained by proteolytic cleavage of a polyanionic component (presumably a sulfated polysaccharide) from the parent acidic form of thrombomodulin, and purified by ion-exchange chromatography. It was previously found that the acidic form of thrombomodulin increases the rate of thrombin inactivation by antithrombin. The present study showed that thrombin bound to acidic thrombomodulin was inactivated at a lower rate by antithrombin in the presence of exogenous heparin than was free thrombin or thrombin bound to the non-acidic form of thrombomodulin. The data suggest that the acidic component of thrombomodulin is primarily responsible for the retardation of thrombin antithrombin complex formation in the presence of exogenous heparin. It is proposed that the polyanionic component of thrombomodulin blocks a site on thrombin required for heparin binding, thus rendering the antithrombin-heparin complex ineffective. PMID- 3028583 TI - [Isolation of human rotaviruses in cultures of rhesus monkey kidney cells]. PMID- 3028584 TI - [Determination of BK virus antibody in different age groups in a Beijing population]. PMID- 3028585 TI - [Effect of fu-ning in treating skeletal fluorosis]. PMID- 3028587 TI - The effects of proteoglycans from different cartilage types on in vitro hydroxyapatite proliferation. AB - Proteoglycans extracted from nasal and articular cartilages have previously been shown to inhibit hydroxyapatite proliferation in vitro. This study now demonstrates that proteoglycans isolated (dissociatively extracted and reaggregated in vitro) from bovine fetal growth plate and bovine occipital condyle, as well as those from bovine nasal cartilage, all retard hydroxyapatite seeded-growth in vitro. On a weight basis (1 mg/ml), the growth plate proteoglycan preparation had a significantly greater inhibitory effect. The greater inhibitory effect of the growth plate proteoglycans as compared to the other cartilage proteoglycan preparations may be related to the unique properties of the proteoglycans in the growth plate, a tissue that undergoes physiological calcification. PMID- 3028586 TI - Effects of the intravenous administration of magnesium sulfate on corrected serum calcium level and nephrogenous cyclic AMP excretion in normal human subjects. AB - The effect of intravenous magnesium sulfate infusion on corrected serum calcium level and parathyroid function assessed by determination of nephrogenous cAMP (NcAMP) excretion were studied in normal human subjects. Significant hypermagnesemia induced by the magnesium sulfate infusion for 120 minutes was accompanied by a gradual and progressive decrease in the corrected serum calcium level. NcAMP excretion fell rapidly, reaching a nadir between 60 and 120 minutes after the infusion began, and after that rose above the baseline excretion. Urinary calcium excretion gradually increased, reaching a peak between 120 and 180 minutes after the infusion began and then gradually decreased. Since magnesium was given as the sulfate, it is not clear whether these changes were attributable to magnesium or sulfate or both. As a control study, we performed intravenous sodium sulfate infusion. The sodium sulfate infusion caused slight hypocalcemia, slight hypercalciuria, and a significant increase in NcAMP excretion. These findings indicate that the hypocalcemia and the hypercalciuria caused by the magnesium sulfate infusion is mainly due to the effect of magnesium, and that the decrease in NcAMP excretion during the infusion is due to the effect of magnesium alone. We conclude that the hypocalcemia caused by the magnesium sulfate infusion is mainly due to the renal calcium loss, and that the inhibition of parathyroid function caused by hypermagnesemia may be only partially involved in the early phase of this hypocalcemia. PMID- 3028588 TI - Treatment of cytomegalovirus retinitis in a patient with AIDS with 9-(1,3 dihydroxy-2-propoxymethyl)guanine. AB - Asymptomatic retinal lesions were found on annual ophthalmic examination in a 38 year-old homosexual man with the acquired immune deficiency syndrome (AIDS). The lesions were compatible with cytomegalovirus retinitis and progressed. Treatment with 9-(1,3-dihydroxy-2-propoxymethyl)guanine (DHPG) resulted in regression of the lesions and improvement in his overall clinical condition. The retinal lesions in AIDS are reviewed. PMID- 3028589 TI - Elimination of human enteric viruses during conventional waste water treatment by activated sludge. AB - The present study was undertaken to determine if viruses were selectively eliminated during waste water treatment. Human enteric viruses were detected at all steps of treatment in a conventional activated sludge waste water treatment plant. Liquid overlays and large volume sampling with multiple passages on BGM cells permitted the detection of poliovirus (serotypes 1, 2, and 3), coxsackievirus B (serotypes 1, 2, 3, 4, and 5), and echovirus (serotypes 3, 14, and 22), as well as reoviruses. The mean virus concentration was 95.1 most probable number of infectious units per litre (mpniu/L) in raw sewage, 23.3 in settled water, 1.4 in effluent after activated sludge treatment, and 40.3 mpniu/L in sludge samples. All samples of raw sewage and settled water, 79% of effluent water, and 94% of sludge samples contained viruses. The mean reduction was 75% after settling and 98% after activated sludge treatment. Poliovirus type 3 was rarely isolated after the activated sludge treatment, but was still detected in about one-third of the sludge samples. Reoviruses and coxsackieviruses were detected at similar rates from all samples and appear to be more resistant to the activated sludge treatment than poliovirus type 3. Poliovirus types 1 and 2 were present in almost every sample of raw sewage and settled water and still found in about half of the effluent and sludge samples, indicating a level of resistance similar to that of reoviruses and coxsackieviruses. PMID- 3028591 TI - Chronic viral diseases. PMID- 3028590 TI - Distribution of extracellular matrix proteins in primary human brain tumours: an immunohistochemical analysis. AB - Using immunohistochemical techniques, we localized several glycoproteins of the extracellular matrix in paraffin-embedded sections of 4 normal brain and 38 primary intracranial tumour specimens. All specimens were positively immunostained to various degrees by monoclonal antibodies to type IV collagen and procollagen III and by antisera to laminin and fibronectin. Staining was consistently most intense at sites of contact between neuroepithelial and mesenchymal or leptomeningeal elements; there was no demonstrable staining within or between neuroepithelial elements in the neuropil. Tumour cells from meningiomas and from the sarcomatous portion of a gliosarcoma were positively immunostained for fibronectin and laminin. The integrity of the glial limitans externa was demonstrated by the positive linear reaction product produced by immunostains for type IV collagen and laminin, even in the most malignant gliomas. The deposition of extracellular matrix glycoproteins at the glial mesenchymal interface observed in this study of primary human brain tumours is a manifestation of one of the interactions between tumour and stromal cells in the central nervous system. A loss of coordination and an alteration in the interactions between epithelial cells and stromal cells across extracellular matrices such as basement membranes are thought to be fundamental steps in the development and progression of cancer. Further characterization studies focusing on other markers of the extracellular matrix are needed to elucidate completely the function of this structure in the central nervous system. PMID- 3028592 TI - Clinical nuclear cardiology: 1. Studies of myocardial perfusion and cellular damage. PMID- 3028593 TI - Malignant fibrous histiocytoma of the heart. A case report and review of the literature. AB - A case report of a 28-year-old woman with malignant fibrous histiocytoma (MFH) of the left atrium is presented, and the six previous reports of this rare cardiac tumor are reviewed. A tendency for malignant fibrous histiocytoma of the heart to occur in the left atrium of young women is suggested; this sarcoma's usual location is in the soft tissue of elderly men. The apparent predilection for the left atrium is unique among cardiac malignancies. Careful pathologic study is necessary to differentiate the uniformly fatal MFH of the heart from the more common benign atrial myxoma. PMID- 3028594 TI - Sudden onset of blindness in patients treated with oral CCNU and low-dose cranial irradiation. AB - Three patients developed the sudden onset of total blindness several months after treatment with oral CCNU and low-dose whole-brain radiation. The anterior visual system was included in the radiation field in all patients. Radiotherapy was given for a frontal-lobe glioblastoma multiforme, for central nervous system prophylaxis in a patient with oat cell carcinoma of the lung, and for a parietal lobe glioblastoma multiforme. None of the neoplasms involved the anterior visual system. The radiation dose ranged from 3000 to 4650 rad and the oral CCNU dosage from 300 mg to 1050 mg. Patients 1 and 2 also received other chemotherapeutic agents. Patient 3 who was treated only with oral CCNU and cranial irradiation died. At autopsy the brain showed a widely infiltrating residual high-grade glioma as well as patchy coagulative necrosis with swollen axons and dystrophic calcifications. The optic chiasm showed severe demyelination, axonal loss, and hyalinized vessels. Synergism between oral CCNU and radiation may account for the blindness produced. PMID- 3028595 TI - Pathology of nasopharyngeal carcinoma. Proposal of a new histologic classification correlated with prognosis. AB - To establish a new histologic classification with better correlation with patient prognosis, the histologic features of nasopharyngeal carcinoma (NPC) were correlated with prognosis and clinical stage among 494 patients who had been followed a minimum of 5 years after initial radiotherapy. A slight modification of World Health Organization (WHO) classification by the separation of spindle cell variant from the nonkeratinizing (NK) and undifferentiated carcinomas (UD) provided a better prognostic correlation: keratinizing squamous cell carcinoma (KS), spindle cell carcinoma (SP), round cell carcinoma (RC), and mixed cell carcinoma (Mix, or NK); 5-year survival rates were 21%, 41%, 51.8%, and 54%, respectively. This prognostic distinction was further improved by dividing the three nonkeratinizing carcinomas (SP, RC, and Mix) into two subtypes each, according to the degree of cell anaplasia and pleomorphism: Type A (with marked anaplasia and/or pleomorphism), and Type B (with moderate or little anaplasia). The three Type A carcinomas had very similar 5-year survival rates (33.3 to 38.6%), as did the three Type B carcinomas (60% to 71.8%). Therefore, a working formulation for the malignancy of NPC emerged: high-grade malignancy (KS; 5-year survival, 21%), intermediate malignancy (Type A carcinomas, 5-year survival, 30% 40%), and low-grade malignancy (Type B carcinomas, 5-year survival rate, 60% 72%). The prognostic distinction remained true after stratification by clinical stage. Therefore, the histologic condition of the tumor of NPC correlated with patient's prognosis. PMID- 3028596 TI - Alternating versus sequential chemotherapy in small cell lung cancer. A randomized German multicenter trial. AB - A total of 306 patients with small cell lung cancer (SCLC) were randomized to receive chemotherapy in a sequential or alternating mode. Sequential chemotherapy consisted of eight cycles of cyclophosphamide, Adriamycin (doxorubicin), and vincristine (CAV) and alternating chemotherapy consisted of three cycles (1, 3, 5) of etoposide, vindesine, and ifosfamide (EVI); three cycles (2, 4, 6) of cisplatin, Adriamycin, and vincristine (PAV); and two cycles (7, 8) of cyclophosphamide, methotrexate, and CCNU (CMC). Responsive patients received prophylactic cranial irradiation after three cycles and chest irradiation after eight cycles of chemotherapy. No maintenance therapy was applied to patients achieving complete remission. Minimum follow-up was 2 years. Of the 302 patients evaluable, overall response rate was 59% in the sequential arm and 70% in the alternating arm. Patients treated with CAV had a complete response rate of 21% in contrast to 36% for those receiving alternating therapy. The median survival for all patients was 9.8 versus 11.3 months, for limited disease 11.1 versus 13.4 months, and for extensive disease 8.9 versus 9.9 months, all in favor of the alternating treatment. Two-year survival rate for all patients was 6% versus 9%, for limited disease 11% versus 14%, and for extensive disease 3% versus 6%, all preferring the alternating treatment mode. Progression-free survival demonstrated a strong correlation to the extent of response irrespective of the treatment regimen applied. Toxicity included 11 lethal and 8 life-threatening complications with a higher frequency in the alternating treatment arm. These results suggest that alternating treatment of SCLC with different drug combinations is more effective than sequential application of CAV. PMID- 3028598 TI - Bilateral adult Wilms' tumor. AB - Bilateral Wilms' tumors in children are not uncommon but in the adult are quite rare. A two-mutation theory of oncogenesis has been proposed in which a prezygotic gene mutation and a postzygotic mutation lead to the development of Wilms' tumor. However, the recent discovery of a postzygotic deletion within the 11p13 band of chromosome 11 in adult cases of Wilms' tumor may explain the rarity of the bilateral form in later years. Early diagnosis is difficult because of the nonspecificity of signs and symptoms. Computer tomography, ultrasonography, and arteriography are important modalities in the earlier diagnosis of adult Wilms' tumor. Because of the rarity of these tumors, no definitive guidelines for treatment have emerged. However, a multidisciplinary approach incorporating surgery, chemotherapy and/or radiation therapy has been used most often but with varying success. This report documents the second case of bilateral adult Wilms' tumor presenting as perinephric hematomas which partially resolved. PMID- 3028597 TI - Epstein-Barr virus and jaw tumors in North Nigeria. AB - Nigerian patients with tumors of the jaw were compared with controls in respect of antibodies to the viral capsid antigens of Epstein-Barr Virus (EBV) and of total immunoglobulin levels. Immunoglobulin levels did not differ between patients and controls but increased two weeks postsurgery. Immunoglobulin A (IgA) antibodies to EBV were detected in a small number of patients, and mean titers of IgG antibodies to EBV were lower in the patient group, indicating a lack of association between EBV and tumor formation. An association was noted between the presence of Hepatitis B surface antigen, and depressed antibody titers to EBV in patients with tumors. Of 78 patients studied, 12% were Hepatitis B surface antigen positive. PMID- 3028599 TI - Familial Hodgkin's disease. A clinical and laboratory investigation. AB - A three to sevenfold increased risk of Hodgkin's disease has been noted in families of patients. We report the first family in which all four of a four member sibship had Hodgkin's disease. In these four sibling cases, we were able to explore markers of infectious etiologic factors by measuring antibodies to Epstein-Barr virus (EBV) and human T-cell lymphotropic virus (HTLV), and possible genetic risk factors by human leukocyte antigen (HLA) typing and chromosome analysis. All three men were diagnosed within six months of their 30th birthday. HLA typing revealed that all four siblings share the DR5 antigen, and two siblings share HLA-B27. Chromosome analysis on peripheral lymphocytes revealed no abnormalities. A detailed, structured interview failed to elicit evidence of unusual exposures. The EBV antibody titers are probably within the normal range and none of the family had antibodies to HTLV-III. Observations on this family suggest that genetic factors play a greater role than environmental factors in the etiology of familial Hodgkin's disease. PMID- 3028600 TI - Immune hemolytic anemia associated with streptococcal preparation OK-432. AB - In the course of administration of an immunopotentiator, streptococcal agent OK 432, a patient with lung cancer was found to have severe anemia and a strongly positive antiglobulin test. In the absence of the drug, eluates from the patient's erythrocytes reacted strongly by the indirect antiglobulin test with a panel of washed Group O erythrocyte samples. The eluated antibody was composed of monoclonal IgG1-lambda. Upon discontinuing the drug the patient's hemoglobin level increased slowly with concomitant normalization of the reticulocyte count. Although very rare, drug-induced immune hemolytic anemia should be included in the list of adverse effects of immunopotentiators. This is the first reported case of immune hemolytic anemia associated with OK-432. PMID- 3028602 TI - Small bowel metastases from primary carcinoma of the lung. AB - Although about half of all patients with carcinoma of the lung have metastases at initial presentation, only nine with metastases to the small bowel have been previously reported. This study was performed to determine the incidence of occult and clinically apparent metastases of lung cancer to the small intestine. Small bowel metastases were present in 46 of 431 patients with primary lung cancer who underwent autopsy during an 11-year period. These patients had an average of 4.8 metastatic sites. Small bowel metastases were present in 12 of 31 (39.0%) patients with large cell carcinoma, 13 of 108 (12.3%) with adenocarcinoma, six of 73 (8.0%) with small cell carcinoma, 15 of 199 (7.5%) with squamous cell carcinoma, and none of 20 with undifferentiated carcinoma. During the same interval, six of 78 patients undergoing small bowel resection for metastatic tumor had lung cancer primaries. Among the nine previously reported clinical cases of small bowel metastases and the six in this series, 14 were operated upon for small bowel perforation and one for obstruction. Nine patients died perioperatively, and no patient survived longer than 16 weeks. These data demonstrate that the incidence of lung cancer metastases to the small bowel is higher than is clinically apparent. Lung cancer metastases to the small bowel often present as intestinal perforation and indicate a poor prognosis; surgery is indicated for palliation. PMID- 3028601 TI - Iron, ferritin, hepatitis B surface and core antigens in the livers of Chinese patients with hepatocellular carcinoma. AB - Surgically resected specimens, consisting of tumor and adjacent non-neoplastic liver tissue, were obtained from 40 patients with primary liver cancer at Zhong Shan Hospital, Shanghai Medical University, the People's Republic of China, between March 1983 and July 1984. All were hepatocellular carcinomas (HCC), one being admixed with cholangiocarcinoma. The relationship of hepatitis B virus (HBV) markers with iron and ferritin was evaluated in liver tissues from patients with primary liver cancers. The serum HBsAg (Hepatitis B surface antigen) positive rate was 80.0% (32/40). Cirrhosis was observed in 97.5% (39/40). HBsAg was identified in 82.5% (33/40) of uninvolved liver, and 35.0% (14/40) of HCC tissues (P less than 0.001). HBcAg (hepatitis B core antigen) was detected in 25.0% (10/40) of liver, and 7.5% (3/40) of HCC tissues (P less than 0.05). Stainable iron was found in 65.0% (26/40) of unaffected livers, and 10.0% (4/40) of HCC tissues (P less than 0.001). Ferritin was demonstrated in 75% (30/40) of non-neoplastic liver, and 40% (16/40) of HCC tissues (P less than 0.001). Twenty two of 33 HCC patients (66.7%) with HBsAg positive cells in their livers also showed stainable iron. Of 16 patients positive for ferritin in HCC cells, iron was found in only two. Iron was found in nine of ten patients with HBcAg in non neoplastic hepatocytes (P = 0.056); a finding compatible with the hypothesis that iron accumulates in cells replicating HBV. The other results indicate that: immunohistologic ferritin in HCC is not due to increased stainable iron; tumor cells may produce ferritin; polyclonal antibodies to human liver ferritin react better with non-neoplastic hepatocytes than with HCC cells; the high prevalence of HBsAg and cirrhosis in HCC suggests that HBV plays a major etiologic role in hepatocarcinogenesis in China; and one case of HCC is attributed to Schistosoma japonicum infestation via cirrhosis. PMID- 3028603 TI - Phase I-II clinical trial of Californium-252. Treatment of stage IB carcinoma of the cervix. AB - Intracavitary Californium-252 combined with whole-pelvis photon radiotherapy was tested as the sole form of treatment for 22 patients with Stage IB carcinoma of the cervix. Californium-252 (Cf) is a fast neutron-emitting radioisotope currently being tested in trials of neutron brachytherapy (NT). The outcomes of the treated group of patients were traced for local tumor control, survival, patterns of failure, and complications. The Cf intracavitary therapy combined with whole-pelvis photon radiotherapy resulted in 95% 2-year and 91% 5-year actuarial survival. There were 9% Grade II-III complications by the Stockholm scale and 4% local failures. These results were obtained in an early clinical trial with a group of largely poor-risk patients with tumors of mean diameter of 4.3 cm. PMID- 3028604 TI - The Na+:H+ antiport in cancer. AB - Several carriers mediate ionic fluxes across the plasma membrane in a variety of mammalian cell types. Intracellular proton concentration is regulated by virtue of the operation of at least two distinct systems: a stilbene-sensitive, Na+- dependent HCO3-/Cl- exchange system, and an amiloride-sensitive Na+/H+ antiporter. The contribution of these two transporters to the modulation of intracellular pH in response to either extracellular pH variations or cell stimulation by growth factors and tumor promoters has been studied in several cell lines, including fibroblast mutants lacking Na+/H+ antiport activity. The attainment of a permissive intracellular pH value is critical to the development of the mitogenic response elicited by growth factors. Kinetic studies have revealed particular features of the Na+/H+ antiporter that explain its function in the early sequence of biochemical events leading to DNA replication. The detailed investigation of the mechanisms by which protons and other ions might regulate cell proliferation has important implications for the understanding of the role of pH microenvironment in carcinogenesis, tumor development and chemotherapy. PMID- 3028605 TI - Binding of polynuclear aromatic hydrocarbons to the rat 4S cytosolic binding protein: structure-activity relationships. AB - The relative competitive binding affinities of benzo[a]pyrene (B[a]P), benzo[e]pyrene, benzo[g, h, i]perylene, picene, 7,12-dimethylbenz [a]anthracene, 1,2,3,4-dibenz[a]anthracene, 1,2,5,6-dibenz[a]anthracene, perylene, 4H cyclopenta[d,e,f]-phenanthrene, benz[a] anthracene, triphenylethylene and triptycene for the rat hepatic cytosolic 4S binding protein were determined using [3H]benzo[a]pyrene as the radioligand. With the exception of triphenlethylene, triptycene and 4H-cyclopenta[d,e,f]phenanthrene, the EC50 values for the remainder of these compounds were between 1.25 X 10(-7) and 2.5 X 10(-8) M with 1,2,5,6-dibenz[a]anthracene being the most active ligand. A comparison of the relative cytosolic Ah (9S) receptor binding affinities and aryl hydrocarbon hydroxylase (AHH) induction potencies of these hydrocarbons with their 4S protein binding affinities demonstrated the following: five compounds, namely 1,2,5,6 dibenz[a]-anthracene, 1,2,3,4-dibenz[a]anthracene, picene, benzo[a]pyrene and 3 methylcholanthrene exhibited high to moderate binding affinities for the 4S and 9S cytosolic proteins (EC50 values less than 10(-6) M) and induced AHH in rat hepatoma cells; three compounds, namely perylene, benzo[e]pyrene and benzo[g,h,i]perylene exhibited high affinities for the 4S binding protein (1.25 X 10(-7), 4.4 X 10(-8) and 2.9 X 10(-8) M, respectively) and low affinities (EC50 values greater than 10(-5) M) for the Ah receptor protein; moreover these three compounds did not induce AHH in rat hepatoma H-4-II E cells in culture. These data suggest that the 4S binding protein may not play a significant role in AHH induction although the results do not rule out a function for this protein in the transregulation of AHH and its associated cytochromes P-450. PMID- 3028606 TI - Retroviruses as carcinogens and pathogens: expectations and reality. AB - Retroviruses (without transforming genes) are thought to cause leukemias and other cancers in animals and humans because they were originally isolated from those diseases and because experimental infections of newborns may induce leukemias with probabilities of 0 to 90%. According to this hypothesis viral cancers should be contagious, polyclonal, and preventable by immunization. However, retroviruses are rather widespread in healthy animals and humans where they typically cause latent infections and antiviral immunity. The leukemia risk of such infections is less than 0.1% and thus about as low as that of virus-free controls. Indeed retroviruses are not sufficient to initiate transformation because of the low percentage of symptomatic virus carriers and the complete lack of transforming function in vitro; because of the striking discrepancies between the long latent periods of 0.5 to 10 years for carcinogenesis and the short eclipse of days to weeks for virus replication and direct pathogenic and immunogenic effects; because there is no gene with a late transforming function, since all genes are essential for replication; because host genes, which do not inhibit virus, inhibit tumorigenesis up to 100% if intact and determine the nature of the tumor if defective; and above all because of the monoclonal origin of viral leukemias, defined by viral integration sites that are different in each tumor. On these bases the probability that a virus-infected cell will become transformed is estimated to be about 10(-11). The viruses are also not necessary to maintain transformation, since many animal and all bovine and human tumors do not express viral antigens or RNA or contain only incomplete proviruses. Thus as carcinogens retroviruses do not necessarily fulfill Koch's first postulate and do not or only very rarely (10(-11)) fulfill the third. Therefore it has been proposed that retroviruses transform inefficiently by activating latent cellular oncogenes by for example provirus integration. This predicts diploid tumors with great diversity, because integration sites are different in each tumor. However, the uniformity of different viral and even nonviral tumors of the same lineage, their common susceptibility to the same tumor resistance genes, and transformation-specific chromosome abnormalities shared with non-viral tumors each argue for cellular transforming genes. Indeed clonal chromosome abnormalities are the only known transformation-specific determinants of viral tumors. Since tumors originate with these abnormalities, these or associated events, rather than preexisting viruses, must initiate transformation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3028608 TI - Coordinate changes in neuronal phenotype and surface antigen expression in human neuroblastoma cell variants. AB - Human neuroblastoma cells growing in culture offer a unique opportunity to study proliferating human cells with a neuronal phenotype. We have previously identified several neuroblastoma cell lines which show spontaneous conversion (N/S interconversion) between two morphologically distinct cell types: neuroblastic (N-type) cells and variant, substrate-adherent (S-type) cells resembling cultured glial or mesenchymal cells. In the present study, we have used molecular markers to confirm the neuronal phenotype of N-type cells and to demonstrate that S-type cells have a nonneuronal phenotype. Furthermore, we have used these markers, including a series of cell surface differentiation antigens, to compare S-type neuroblastoma cells with a wide range of cultured epithelial, mesenchymal, and neuroectodermal cells. The results suggest that N/S interconversion represents an ordered transition between two neuroectodermal differentiation programs rather than random phenotypic instability of cultured cells; S-type variant cells show a molecular phenotype most closely resembling the phenotype of cultured ectomesenchymal cells; and in vitro variant formation of human neuroblastomas may provide an experimental model for the observed in vivo transition of some malignant neuroblastomas into benign ganglioneuromas. PMID- 3028607 TI - HL-60-1E3, a novel phorbol diester-resistant HL-60 cell line. AB - The HL-60 (human promyelocytic leukemia) cell line has proved to be a useful model for the study of the expression of cellular functions and markers associated with hematopoietic differentiation. We report here the development and initial characterization of a novel, differentiation-resistant HL-60 cell line (HL-60-1E3) which was established by cloning the parent HL-60 line in the absence of mutagens or differentiation-inducing agents. HL-60-1E3 exhibits markedly reduced phorbol diester-induced expression of extracellular cytolytic activity, nonspecific esterase, phagocytosis, and surface Mo1 antigen. In addition, dimethyl sulfoxide-induced expression of both Mo1 and Mo2 is markedly reduced. However, if HL-60-1E3 is exposed to dimethyl sulfoxide, it acquires appreciable phorbol diester-triggered cytolytic activity and production of superoxide anion (O2-). Phorbol diester receptor number and dissociation constant (Kd) obtained by Scatchard analysis are not significantly different for the two cell lines. The HL 60-1E3 cell line should provide a useful adjunct to other cell lines used in the study of normal myeloid and leukemic cell differentiation, as well as the study of cytokine maturation factors and oncogene expression. PMID- 3028609 TI - An animal model for colon cancer metastasis: establishment and characterization of murine cell lines with enhanced liver-metastasizing ability. AB - A detailed understanding of the pathogenesis of colon cancer metastasis has been hindered by the lack of appropriate animal models which accurately reflect events in this complex process. An animal model for colon cancer metastasis is described in which spontaneously metastasizing colonic tumors are formed after injection of murine colon cancer cells into the cecal wall of BALB/c mice. Using this model, tumor cells with different liver-metastasizing potential were selected and shown to possess several properties known to be associated with other metastatic cell lines. The ability of tumor cells to invade a reconstituted basement membrane and to secrete type IV collagenase was directly proportional to their metastatic ability. In addition, liver-metastasizing cells preferentially migrated toward liver extracts in a Boyden chamber assay, as compared to extracts of brain or lung, and adhered rapidly to highly purified hepatic sinusoidal endothelial cells versus hepatic parenchymal cells in vitro. This model may thus be useful for studying many aspects of the pathogenesis of colon cancer metastasis. PMID- 3028610 TI - Secretion of type IV collagenolytic protease and metastatic phenotype: induction by transfection with c-Ha-ras but not c-Ha-ras plus Ad2-E1a. AB - Activated ras oncogene transfection into suitable recipient cells has been shown to induce the metastatic phenotype (Thorgeirsson, et al., Mol. Cell. Biol., 5: 259-262, 1985). We have used this model system to study the correlation of basement membrane collagenolysis with metastatic propensity. The c-Ha-ras oncogene alone, or combined with v-myc, transfected into early passage rat embryo fibroblasts, induce these cells to secrete high levels of type IV collagenolytic metalloproteinase and to concomitantly exhibit a high incidence of spontaneous metastases in nude mice. Cotransfection of c-Ha-ras plus the adenovirus type 2 E1a gene yields cells which are highly tumorigenic but nonmetastatic and fail to produce type IV collagenase. This effect is due to a suppression of collagenase elaboration, not increased production of a collagenase inhibitor, and not decreased production of a collagenase activator. The characteristics of the collagenase are identical to tumor type IV collagenase described previously. The nonmetastatic cells which failed to produce type IV collagenase retain the ability to secrete high levels of plasminogen activator. Transfection with the protooncogenic forms of Ha-ras or mos, or spontaneous transformation of NIH 3T3 cells or chemical transformation of BALB 3T3 cells yields cells which fail to produce collagenase, are tumorigenic, but totally nonmetastatic. These data support a biochemical linkage of type IV collagenase expression with the metastatic phenotype in this rodent system. PMID- 3028611 TI - Production of transforming growth factors by simian sarcoma virus-transformed cells. AB - Cellular transformation of normal rat kidney (NRK) cells by simian sarcoma virus (SSV) results in a complete loss of the cellular requirement of externally added polypeptide growth factors for proliferation. Moreover, SSV-transformed NRK cells have a strongly reduced ability to bind both external platelet-derived growth factor and epidermal growth factor, when compared to nontransformed NRK cells. Analysis of serum-free medium conditioned by SSV-transformed NRK cells shows that this cell line secretes both types alpha and beta transforming growth factor (TGF). The level of TGF alpha production (300 ng/liter conditioned medium) by SSV transformed NRK is among the highest described to date. Since addition of TGF alpha and beta in combination is sufficient to induce phenotypic transformation of NRK cells, it is concluded that although expression of the sis oncogene is essential for transformation, expression of additional genes may be required for the phenotypic alterations accompanying complete cellular transformation. PMID- 3028612 TI - Generation of superoxide (O2-.) from alveolar macrophages exposed to asbestiform and nonfibrous particles. AB - Active oxygen species are implicated causally in tumor promotion by 12-O tetradecanoylphorbol-13-acetate and in cell damage by asbestos, a putative tumor promoter of the respiratory tract. To determine the properties of asbestos important in generation of the oxygen free radical, superoxide (O2-.), hamster and rat alveolar macrophages were exposed in vitro to nontoxic concentrations of fibrous (crocidolite, erionite, Code 100 fiberglass, sepiolite) and nonfibrous (riebeckite, mordenite, glass) particulates. The amount of O2-. released by cells in response to dusts was determined by measuring the reduction of cytochrome c in the presence and absence of superoxide dismutase. Results showed that all fibrous (defined as a greater than 3:1 length:diameter ratio) dusts caused a significant increase in both release of O2-. from rat macrophages and enhancement of zymosan triggered O2-. from hamster macrophages. Nonfibrous particles were less active than fibers at comparable concentrations. These results suggest that the geometry of particulates is of critical importance in the generation of O2-. from cells of the respiratory tract. PMID- 3028613 TI - Multifactorial drug resistance in an adriamycin-resistant human small cell lung carcinoma cell line. AB - In a human small cell lung carcinoma cell line, GLC4, Adriamycin (ADR) resistance was induced. In the resistant cell line, GLC4/ADR, a 45% decreased intracellular ADR level was found compared to GLC4, but this could not fully explain the resistance. Evaluation of DNA damage in both cell lines after incubation with ADR by alkaline and neutral elution revealed single-strand breaks, DNA-protein cross links, and double-strand breaks (DSB). At all incubation concentrations there was a decreased amount of all types of DNA damage in GLC4/ADR. The number of DSB was decreased also when corrected for the decreased intracellular concentration. This can at least partly be explained by the decreased stability of ADR induced DSB. After removal of ADR, 80% of DSB was repaired in 1 h in GLC4/ADR against no repair in GLC4. X-ray induced DSBs were also repaired faster: in GLC4/ADR t1/2 = 10 min and in GLC4 t1/2 = 23 min. Ratios for single strand breaks/DSBs and single strand breaks/DNA-protein cross-links between GLC4 and GLC4/ADR after exposure to ADR differed; these differences were compatible with differences in the distribution of the various types of DNA damage induced in the cell lines due to an altered ADR-topoisomerase II interaction. In this human small cell lung carcinoma cell line the resistance is multifactorial with decreased intracellular ADR levels, increased DNA repair, and altered ADR-topoisomerase interaction. PMID- 3028614 TI - Relationship between the intracellular effects of camptothecin and the inhibition of DNA topoisomerase I in cultured L1210 cells. AB - Results of filter elution assays of lesions produced in the DNA of cultured L1210 cells by the antineoplastic alkaloid camptothecin support the notion that topoisomerase I is an intracellular target of this drug. One to 10 microM camptothecin induced DNA single-strand, but not double-strand, breaks when incubated with intact cells or with their isolated nuclei. Approximately one half of the strand breakage was protein concealed, as judged by filter elution. Camptothecin-induced, protein-concealed DNA strand breaks disappeared rapidly after drug removal. DNA-protein cross-links were generated by camptothecin with frequencies approximately equal to those of protein-concealed DNA strand breaks. It is likely that camptothecin can inhibit topoisomerase I in intact cells in a manner similar to that in which other antineoplastic agents such as amsacrine or teniposide inhibit topoisomerase II. DNA-breaking lesions other than those resulting from trapped topoisomerase I-DNA complexes may also be generated by camptothecin. The yields of DNA strand breaks induced by camptothecin, amsacrine, or teniposide were approximately doubled when cells were incubated for 16 h with 3-aminobenzamide, an inhibitor of poly(ADP ribosylation) of proteins, prior to 1 h exposure to the antineoplastic compounds. 3-Aminobenzamide also enhanced the cytotoxic action of camptothecin, amsacrine, and teniposide. These results suggest that protein-concealed strand breaks can be lethal lesions and that intracellular topoisomerase I and II activity may be regulated coordinately through poly(ADP ribosylation). PMID- 3028615 TI - Role of uridine triphosphate in the phosphorylation of 1-beta-D arabinofuranosylcytosine by Ehrlich ascites tumor cells. AB - Pyrimidine nucleotide pools were investigated as determinants of the rate of phosphorylation of 1-beta-D-arabinofuranosylcytosine (ara-C) by Ehrlich ascites cells and cell extracts. Cells were preincubated for 2 h with 10 microM pyrazofurin, 10 mM glucosamine, 50 microM 3-deazauridine, or 1 mM uridine in order to alter the concentrations of pyrimidine nucleotides. Samples of the cell suspensions were taken for assay of adenosine 5'-triphosphate (ATP), uridine 5' triphosphate (UTP), cytidine 5'-triphosphate, guanosine 5'-triphosphate, deoxycytidine 5'-triphosphate (dCTP), and deoxythymidine 5'-triphosphate; then 1 microM [3H]ara-C was added and its rate of intracellular uptake was measured for 30 min. 3-Deazauridine lowered dCTP and stimulated ara-C uptake; however, pyrazofurin and glucosamine were potent inhibitors of ara-C uptake although they also decreased dCTP levels. Uridine stimulated ara-C uptake despite an increase in dCTP. A crude cytoplasmic extract was prepared by a procedure which permitted results of ara-C kinase assays to be expressed as pmol per min per 10(6) cells as in the cellular uptake studies. When assayed in the presence of mixtures of ribo- and deoxyribonucleoside triphosphates at concentrations close to their cellular levels, ara-C kinase activity closely approximated the cellular uptake rate for the five incubation conditions. Deletion of cytidine 5'-, guanosine 5'-, or deoxythymidine 5'-triphosphate from the assay mixture had little effect, while deletion of dCTP increased kinase activity 9-fold. Elimination of ATP also did not alter kinase activity in the presence of the remaining five ribo- and deoxyribonucleoside triphosphates; however, deletion of UTP reduced activity to 22% of the rate with the control mixture. When ara-C kinase was assayed with only 3 mM ATP, dCTP was a very potent inhibitor (50% inhibition concentration = 0.4 microM). Inhibition was complete at 10 microM dCTP, a concentration below the intracellular dCTP level in control cells (25 microM). With 0.9 mM UTP, enzyme activity was 2-fold greater in the absence of dCTP and the dCTP was 15-fold less potent as an inhibitor (50% inhibition concentration = 6 microM). We conclude that the actual phosphate donor for the phosphorylation of 1 microM ara-C in Ehrlich cells is UTP and not ATP. These observations suggest that successful combination protocols aimed at stimulating ara-C uptake by means of a decrease in dCTP levels must simultaneously preserve or increase UTP pools. PMID- 3028616 TI - Direct effects of the pyrimido-pyrimidine derivative RA 233 (Rapenton) on rat 13762NF mammary tumor cell clones in vitro. AB - Studies with the pyrimido-pyrimidine analogue RA 233 (Rapenton) suggest that its antimetastatic action may not be mediated entirely by inhibition of platelet function. Little is known about its direct effects on tumor cells. We investigated the in vitro effects of RA 233 on clones MTLn3 and MTC of differing metastatic potentials, isolated from the 13762NF rat mammary adenocarcinoma. The results indicated that RA 233 is cytostatic (EC50 of approximately 140 microM and approximately 180 microM for MTLn3 and MTC cells, respectively) rather than cytotoxic by determining changes in viable cell number, thymidine uptake, and incorporation of thymidine and methionine. In both clones RA 233 inhibited cAMP dependent phosphodiesterase activity and affected cAMP accumulation in intact cells. In contrast, clonal heterogeneity in drug-induced morphological changes, such as vacuole formation and altered organization of cytoskeletal structures, as well as increased tumor cell growth at 50 microM RA 233 was observed between clones MTLn3 and MTC. These data could explain the conflicting results obtained with RA 233 when evaluated as an antimetastatic agent. PMID- 3028617 TI - Characterization of two cell lines with distinct phenotypes established from a patient with small cell lung cancer. AB - Two small cell lung cancer (SCLC) cell lines were established from pericardial and pleural effusions of a patient with histopathologically proven SCLC of the oat cell type. Chemotherapy was administered without response during the 148-day period prior to the establishment of the first cell line, SCLC-22H, and some of the same drugs were administered in the 15 days prior to the establishment of the second cell line, SCLC-21H. Both cell lines differed markedly in their biochemical, kinetic, and morphological properties. Although the biomarkers L Dopa decarboxylase, bombesin, carcinoembryonic antigen, and neurotensin were detectable in SCLC-22H, they were undetectable or low in SCLC-21H. The population doubling time was twice as fast and the colony forming efficiency higher in SCLC 21H than in SCLC-22H. They both expressed high concentrations of the SCLC marker enzymes neuron-specific enolase and the creatine kinase isoenzyme BB and showed no significant differences in their chromosomal characteristics. c-myc was amplified and expressed in both cell lines, and SCLC-21H had an additional rearranged and amplified EcoRI c-myc fragment. Morphological differences were apparent in liquid culture, cytology, and xenograft histology; SCLC-21H grew much more loosely than SCLC-22H, and had more prominent nucleoli and more abundant cytoplasm. Ultrastructurally dense core granules were present in both cell lines. SCLC-21H thus expressed properties of the variant cell type of SCLC, whereas SCLC 22H had mixed classic/variant features. An in vivo progression of the patient's tumor from a pure small cell to a mixed small cell/large cell morphology could be demonstrated, which suggests that cell line SCLC-22H represents a cell type characteristic for the transitional phase of the tumor. The features of this cell line therefore define a new subclass of SCLC called transitional cell type. SCLC 22H may be of use to study the mechanisms involved in the classic to variant transition, which probably has a considerable impact on the prognosis and response to therapy. PMID- 3028618 TI - Selective anticancer effects of 3',5'-dioctanoyl-5-fluoro-2'-deoxyuridine, a lipophilic prodrug of 5-fluoro-2'-deoxyuridine, dissolved in an oily lymphographic agent on hepatic cancer of rabbits bearing VX-2 tumor. AB - 3',5'-Dioctanoyl-5-fluoro-2'-deoxyuridine (FdUrd-C8), one of the lipophilic prodrugs of 5-fluoro-2'-deoxyuridine (FdUrd) was dissolved in an oily lymphographic agent (Lipiodol Ultra-Fluid), which had been studied as a carrier of the anticancer drug for hepatic cancer. The prodrug was administered into the left proper hepatic artery of rabbits bearing VX-2 tumor in the liver in order to examine the anticancer effects and possible adverse effects on nontumorous hepatic cells. Lipiodol or FdUrd-C8*Lipiodol selectively remained in the hepatic cancer area but disappeared from nontumorous parts of the liver 7 days after injection. Tumor growth rates in 1 week of the untreated group, a group given injections of 0.2 ml of Lipiodol alone, and groups given injections of 0.2 ml of Lipiodol containing 30, 50, 70, and 100 mg of FdUrd-C8 were 636, 436, 34.8, 14.9, -2.4, and -10.4% of the size at the time of treatment, respectively. Pathological observation also showed that FdUrd-C8 had a strong anticancer effect on VX-2 tumor growing in the liver of the rabbits. In contrast to the effect on the cancerous cells, that on nontumorous hepatic cells was very slight. In pathological observation, necrosis or degeneration of nontumorous hepatic cells was hardly observed. Plasma glutamic-oxaloacetic transaminase and glutamic pyruvic transaminase levels temporarily rose 1 day after injection but returned to the initial levels within 7 days in all groups. PMID- 3028619 TI - Phase I study of epirubicin given on a weekly schedule. AB - Epirubicin was studied in a phase I setting to find the maximum tolerated dose when given weekly for 3 of 4 weeks. Forty-one evaluable patients were treated in groups at doses increasing from 20 to 45 mg/m2. The highest dose level produced the maximum degree of myelosuppression (lowest neutrophil count, 1.9 X 10(9)/L; range, 0-3.7) recorded on Day 22. This was well-tolerated in this group of mainly pretreated patients. Nonhematologic side effects were minimal. This dose schedule allows a greater dose per unit time to be administered than other recommended schedules for epirubicin. PMID- 3028621 TI - Phase II study of carboplatin in previously untreated patients with metastatic small cell lung carcinoma. AB - Fourteen previously untreated patients with metastatic small cell lung carcinoma (SCLC) were treated with carboplatin, 400 mg/m2, given as a 1-hour infusion every 28 days. Eleven patients had a partial response to therapy [response rate, 79% (95% confidence limits, 57%-100%)]. The median duration of response was 8 weeks (range, 4-41) and the median survival was 29 weeks (range, 11-68+). Hematologic toxicity was limited to one patient who developed World Health Organization (WHO) grade IV thrombocytopenia; no patient developed infection due to granulocytopenia. Nonhematologic toxicity was limited to WHO grade I and II nausea and vomiting. This study demonstrates that carboplatin is a highly active drug in SCLC with minimal toxicity. Future studies should evaluate its use in combination with other chemotherapeutic drugs in SCLC. PMID- 3028620 TI - Small cell carcinoma of the lung: influence of age on treatment outcome. AB - Two hundred twenty-three patients between 29 and 78 years of age with small cell cancer of the lung were studied in serial protocols at the University of Maryland Cancer Center from 1975 to 1982. Each of these patients received chemotherapy consisting of doxorubicin, cyclophosphamide, and etoposide. In this paper we explore age and its interaction with the patient's extent of disease as it relates to clinical outcome variables such as the achievement of a complete response (CR), survival times, and various measures of toxicity. Preliminary evidence is found to suggest that an age-extent of disease interaction affects both the probability of obtaining a CR and the length of survival. Patients of all ages can benefit from treatment of small cell carcinoma of the lung, with CR as the treatment goal. However, increased toxicity and early death remain as barriers to successful treatment of the elderly. PMID- 3028623 TI - [Role of angiotensin converting enzyme inhibitors in the therapy of hypertension]. PMID- 3028622 TI - Phase II study of aclarubicin in non-small cell lung cancer. PMID- 3028624 TI - [Effects of enalapril on the mass and function of the left ventricle in hypertensive subjects]. PMID- 3028625 TI - [Clinical experience with enalapril in the treatment of essential mild-moderate arterial hypertension]. PMID- 3028626 TI - [Combination of enalapril with diuretics in the treatment of arterial hypertension]. PMID- 3028627 TI - [Long-term experience with enalapril]. PMID- 3028628 TI - [Physiopathologic bases for the use of angiotensin converting enzyme inhibitors in congestive heart failure]. PMID- 3028629 TI - [Clinical experience with enalapril in the treatment of heart failure]. PMID- 3028630 TI - [Changes in the variability of the heart rate induced by controlled respiration and beta-adrenergic block]. PMID- 3028632 TI - Transcatheter arterial embolization in hepatoma complicated with obstructive jaundice. AB - Transcatheter arterial embolization was performed on a patient with hepatoma complicated by obstructive jaundice after the patient's condition had been improved by percutaneous biliary drainage. As a result of the embolization, a reduction in size was observed in both the main tumor and the tumor that had invaded the common bile duct. Even after removing the biliary drainage tube, there was no recurrence of obstructive jaundice for 6 months. PMID- 3028631 TI - [Hypotensive effect of ketanserin and evaluation of its alpha-blocking activity]. PMID- 3028634 TI - Sodium monofluorophosphate degradation by oral streptococci, plaque and saliva. PMID- 3028633 TI - A multicenter trial of enalapril in the treatment of essential hypertension. AB - The therapeutic profile of enalapril in mild to moderate uncomplicated essential hypertension was assessed in 265 patients who participated in a multicenter, open label, prospective study lasting eight weeks. There were 54 younger (aged 39 years or less), 136 middle-aged (40 to 59 years), and 75 older patients (60 years or over). Monotherapy with enalapril in a single daily dosage regimen ranging between 5 and 40 mg resulted in normotension (in the sitting position) in 73% of the younger, 50% of the middle-aged, and 56% of the older patients. Normotension was achieved with 5 mg/day of enalapril in 41%, 18%, and 37% of the subgroups, respectively. Both systolic and diastolic pressures at the end of eight weeks of treatment were significantly lower (P less than 0.01) in the younger patients than in the other two age groups. White patients had significantly greater (P less than 0.001) response of both systolic and diastolic blood pressures than did black patients and required significantly smaller (P less than 0.01) average daily dosages of enalapril (14 mg versus 22 mg, respectively). The overall incidence of side effects was 14% among all 276 patients enrolled in the study. Most were mild and transient, but six patients discontinued enalapril during the first week of therapy because of side effects. There were no cases of rash, dysgeusia, hematological disorders, or deterioration in renal function, but there were two cases of pruritus, one of glossitis associated with an upper respiratory infection, and three of dry cough or wheezing. Angioedema was not observed. Monotherapy with enalapril, usually in a single daily dose of 10 to 20 mg, was effective in inducing normotension in approximately half of the middle-aged and older hypertensive individuals and in nearly three fourths of those below age 40. In this study it was generally well tolerated, with a relatively small incidence of side effects. PMID- 3028636 TI - Immunocytochemical localization of lactoferrin in human neutrophils. An ultrastructural and morphometrical study. AB - The subcellular localization of lactoferrin in human neutrophils was studied by an electron-microscopic immunoperoxidase method. This molecule was detected in small granules of blood polymorphonuclear leukocytes. A morphometrical analysis showed that there was no significant difference in the mean size between lactoferrin-positive and myeloperoxidase-negative granules. In contrast, the mean size of myeloperoxidase-positive granules was significantly larger than that of lactoferrin-positive granules. This indicates that lactoferrin is contained in the myeloperoxidase-negative, secondary, granules of human neutrophils. In immature bone marrow mononuclear neutrophils, lactoferrin was present in cytoplasmic granules of somewhat larger size than lactoferrin-positive granules of polymorphonuclear leucocytes. A morphometrical study showed that the mean size of lactoferrin-positive granules was significantly greater in immature bone marrow cells than in polymorphonuclear leucocytes. This indicates that lactoferrin-positive granules decrease in size as the cells mature. Besides cytoplasmic granules, lactoferrin was demonstrated in the Golgi complex and a part of the rough endoplasmic reticulum of immature bone marrow neutrophils, probably myelocytes and early metamyelocytes. These results show that lactoferrin is synthesized and packed into secondary granules in immature bone marrow neutrophils and therefore that the secondary granules are a type of secretory granule. PMID- 3028635 TI - [Antibodies against capsid (VCA) and nuclear (EBNA) antigens of the Epstein-Barr virus in patients with rheumatoid arthritis]. PMID- 3028638 TI - [Investigations on the antibody against adult diarrhoea rotavirus among the healthy population in China]. PMID- 3028637 TI - Calspermin: a 32K-dalton calmodulin-binding protein in central nervous system and male reproductive system; property and distribution. PMID- 3028639 TI - Hepatitis BeAg in chronic asymptomatic hepatitis B surface antigen carriers and in primary hepatocellular carcinoma patients. PMID- 3028640 TI - Integrins: a family of cell surface receptors. PMID- 3028641 TI - The inducible lac operator-repressor system is functional in mammalian cells. AB - We have investigated the use of the Escherichia coli lac operator-repressor system to regulate expression of transfected genes in mammalian cells. We show that lac repressor produced in mouse L cells by transfection of a lacl expression vector blocks transcription of an MSV-CAT fusion gene when the lac operator is inserted at any one of the following sites within the promoter region: between the initiation codon (ATG) and the transcription start site; between the transcription start and TATA box regions; or upstream of the TATA box region. This last result suggests that the repressor may prevent protein-protein interactions involved in transcription activation. The inducer IPTG causes a marked derepression of CAT expression. The lac repressor-operator complex may be useful as an on/off "switch" in the regulation of gene expression for gene transfer experiments. PMID- 3028642 TI - Both positive and negative regulators of HO transcription are required for mother cell-specific mating-type switching in yeast. AB - The HO gene, which encodes an endonuclease responsible for initiating mating type switching in yeast, is transcribed at START during the cell cycle of mother cells but not at all during the cell cycle of daughter cells. At least six genes, called SWI1-6, are necessary for HO transcription. We describe the isolation and characterization of mutations in two new genes called SDI1 and SDI2, which partially suppress the requirement for SWI5 and which cause daughter cells to express HO. The analysis of mating type switching in swi5- sdi1- and SWI5+ sdi1- strains suggests that the mother cell specificity of HO transcription is due exclusively to the selective action of SWI5 in mother cells. SDI1 encodes (or regulates) a repressor protein that binds to the HO promoter and prevents HO transcription in daughter cells by causing HO to be fully SWI5 dependent. PMID- 3028643 TI - Translation is required for regulation of histone mRNA degradation. AB - When DNA synthesis is inhibited, the mRNAs coding for the replication-dependent histone proteins are selectively destabilized. The histone genes have been altered and reintroduced into tk- mouse L cells by cotransfection with the herpesvirus thymidine kinase gene. Two features of the mRNA are necessary for regulation of degradation: first, the hairpin loop must be present at the 3' end of the histone mRNA; and second, the histone mRNA must be capable of being translated to within 300 nucleotides of the 3' end of the RNA. Polyadenylated histone mRNAs are stable, as are histone mRNAs that contain in-frame termination codons early in the coding region or 500 nucleotide 3' untranslated regions with a normal hairpin loop at the 3' end. PMID- 3028644 TI - Translational regulation of SV40 early mRNA defines a new viral protein. AB - SP6-initiated in vitro transcripts, representing the three major classes of early SV40 mRNAs, early-early (EE) and two late-early (LE) transcripts, were assayed by in vitro translation to compare their relative efficiencies for synthesis of the SV40 T antigens. The presence of one or two potential AUG initiator codons in the leader sequences of the LE RNAs inhibits efficient translation from the downstream T-antigen initiator AUG. In vitro translation of the capped form of the shorter SV40 LE RNA resulted in the synthesis of a 2.7-kd protein. In vivo pulse labeling of SV40-infected CV-1 cells demonstrated the accumulation of a peptide of similar size at late times after lytic infection, indicating that it is an authentic viral protein encoded by the early leader sequence of SV40. PMID- 3028645 TI - Structure-function analysis of fos protein: a single amino acid change activates the immortalizing potential of v-fos. AB - We have undertaken a detailed structure-function analysis of v-fos protein, taking the ability to induce transformation and immortalization as criteria of biological activity. Our results demonstrate that the evolutionarily conserved center region of fos, comprising ca. 28% of the protein, is indispensable for its function. A single amino acid change (Glu 138----Val 138) in this region activates the immortalizing potential of v-fos without affecting its transforming capacity. The presence of additional either N- or C-terminal amino acids, however, is required for efficient expression of a stable protein, and significantly increases its biological activity. In contrast, sequences in the C terminal half (between positions 228 and 267) strongly downmodulate the transforming activity of v-fos protein without decreasing its immortalizing potential. PMID- 3028646 TI - Cloning of the beta subunit of the leukocyte adhesion proteins: homology to an extracellular matrix receptor defines a novel supergene family. AB - We have isolated cDNA clones encoding the beta subunit of the human LFA-1, Mac-1, and p150,95 family of leukocyte adhesion proteins. The deduced 769-amino-acid sequence defines a cysteine-rich polypeptide with the characteristic features of an integral membrane protein. Peptide sequence data, Northern blot analysis, and Southern blot analysis suggest that a single gene encodes the beta subunit of all three leukocyte adhesion proteins. There is 45% homology between the beta subunit sequence and band III of integrin, a chick fibronectin and laminin receptor. This homology defines a new supergene family of cellular adhesion proteins. PMID- 3028647 TI - Deletion analysis of GAL4 defines two transcriptional activating segments. AB - We describe the activities of a wide array of deletion mutants of GAL4, a yeast transcriptional activator. We identify two short regions of GAL4, each of which activates transcription when fused to the DNA-binding region of the molecule. Very large portions of GAL4 are not required for gene activation. PMID- 3028648 TI - Distinct sequence determinants direct intracellular sorting and modification of a yeast vacuolar protease. AB - We have mapped a sequence determinant in the vacuolar glycoprotein carboxypeptidase Y (CPY) that directs intracellular sorting of this enzyme. Through the study of hybrid proteins, consisting of amino-terminal segments of CPY fused to the secretory enzyme invertase, we have found that the N-terminal 50 amino acids of CPY are sufficient to direct delivery of a CPY-Inv hybrid protein to the yeast vacuole. Our data suggest that this 50 amino acid segment of CPY contains two distinct functional domains; an N-terminal signal peptide followed by a segment of 30 amino acids that contains the vacuolar sorting signal. Deletion of this putative vacuole sorting signal from an otherwise wild-type CPY protein leads to missorting of CPY. Furthermore, examination of the Asn-linked oligosaccharides present on CPY and CPY-Inv hybrid proteins suggests that an additional determinant in CPY specifies the extent to which these proteins are glycosylated in the Golgi complex. PMID- 3028650 TI - Effects of inhibitors of C1q biosynthesis on macrophage Fc receptor subclass mediated antibody-dependent cellular cytotoxicity and phagocytosis. AB - We examined the effects of the inhibitors of C1q or collagen biosynthesis, 2,2' dipyridyl (DP), and 3,4-dehydro-DL-proline (DHP) on murine macrophage (M phi) FcR subclass-mediated antibody-dependent cellular cytotoxicity (ADCC) and phagocytosis of sheep erythrocyte targets. Oil-elicited peritoneal M phi from C3HeB/FeJ mice which were cultured for 24 hr with DP (0.08 or 0.10 mM) or DHP (0.8 or 1.0 mM) showed a significant decrease in FcR subclass-mediated ADCC for murine monoclonal IgG2a (FcRI) and IgG2b/IgG1 (FcRII) as well as for heterologous polyclonal IgG. These collagen inhibitors also blocked phagocytosis mediated by both IgG2a- and IgG2b-opsonized erythrocytes. DP was more potent than DHP in blocking FcR effector functions in a reversible fashion and neither inhibitor affected M phi C3b receptor function. Pretreatment of M phi with collagenase resulted in significant reduction in FcR-mediated ADCC and phagocytosis. The inhibition of M phi FcR subclass-mediated ADCC and phagocytosis by collagen C1q synthetic inhibitors or by collagenase treatment further confirms a functional relationship between cell-associated C1q and FcR-dependent functions. PMID- 3028649 TI - Protein sorting in yeast: the localization determinant of yeast vacuolar carboxypeptidase Y resides in the propeptide. AB - We have isolated cis-acting mutations in the gene encoding the yeast vacuolar protein carboxypeptidase Y (CPY) that result in missorting and aberrant secretion of up to 95% of newly synthesized CPY. The CPY polypeptides synthesized by these mutants use the late secretory pathway to exit the cell, since the late-acting sec1 mutation prevents their secretion. The mutant versions of CPY are secreted as the proCPY zymogen and are enzymatically activatable in vivo and in vitro. All the mutations, including small deletions and an amino acid substitution, map to the amino-terminal propeptide region and define a discrete yeast vacuolar localization domain whose integrity is required for efficient sorting of the CPY zymogen. Thus, the N-terminal propeptide of CPY carries out at least three functions: it mediates translocation across the endoplasmic reticulum, renders the enzyme inactive during transit, and targets the molecule to the vacuole. PMID- 3028652 TI - Glutathione conjugate formation without N-demethylation during the peroxidase catalysed N-oxidation of N,N',N,N'-tetramethylbenzidine. AB - The mechanism of peroxidative N-dealkylation of alkylamines proceeds via one electron oxidation to the iminium cation which reacts with water to give the N hydroxymethyl derivative which decomposes to formaldehyde and the N-demethylated product. This reaction is normally inhibited by glutathione by reduction of the cation radical with subsequent formation of oxidized glutathione (GSSG) with oxygen uptake. It was found that the horseradish peroxidase catalyzed N demthylation of N,N,N',N'-tetramethylbenzidine (N4-TMB) in the presence of glutathione leads to the formation of water-soluble metabolites identified by high field nuclear magnetic resonance (NMR) and fast atom bombardment (FAB) mass spectrometry as 3,3'-(diglutathion-S-yl) and 2,2'-(diglutathion-S-yl)-N4-TMB. Smaller amounts of (monoglutathion-S-yl)-N4-TMB were also found. Only trace amounts of GSSG were formed and no oxygen uptake was observed. Electron spin resonance (ESR) spectrometry in the presence of 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) did not indicate the presence of a DMPO-glutathionyl adduct. These results indicate that glutathione inhibited the N-demethylation of N4-TMB under the described reaction conditions not by reduction of the cation radical but by conjugate formation. The mechanism of N-demethylation must involve removal of two successive electrons to give the benzoquinone-diimine which undergoes rearrangement to the iminium cation followed by reaction with water. PMID- 3028651 TI - Differential requirement for accessory cells in polyclonal T-cell activation. AB - Highly purified rat Ia-negative (OX-6-) and Ia-positive (OX-6+) T cells were employed to examine the requirement for accessory cells (AC) and/or soluble factors in the activation of resting T cells with Con A, PHA, sodium periodate, or antigen. A variety of cells were employed as AC, including Ia-positive and Ia negative macrophages (M phi), gamma-irradiated (2000 rad) or non-irradiated OX-6+ T cells, and several Ia-negative adenovirus-transformed rat embryo fibroblast cell lines. Our results suggested that for the expression of IL-2 receptors (IL 2R) and proliferation of OX-6- T cells in response to Con A, PHA, or antigen, there was an obligatory requirement for the presence of AC which could not be overcome by the addition of IL-1 and/or IL-2. Activation of OX-6- T cells with antigen required the presence of Ia+ AC, while activation with mitogens could be initiated with Ia- AC. M phi were efficient in AC function in all responses tested, while the AC function of OX-6+ T cells (TAPC) proved discriminatory under different conditions. The optimal response to PHA required much higher concentrations of TAPC as AC than for the Con A response. TAPC failed to stimulate sodium periodate-treated T cells under any conditions tested. Furthermore, when TAPC were employed as AC, their antigen-presenting ability was radiosensitive, while their AC function for Con A and PHA was radioresistant. These results suggest that molecules involved in T cell-AC interactions may differ, depending on the source of AC and/or type of the proliferative stimulus provided to T cells. This data has been discussed in the context of T-cell activation. PMID- 3028653 TI - Synthesis and angiotensin converting enzyme-inhibitory activity of 1,5 benzothiazepine and 1,5-benzoxazepine derivatives. III. PMID- 3028654 TI - Preventive effect of 2-halogen-substituted derivatives of cyclic adenosine 3',5' monophosphate on experimental disseminated intravascular coagulation. PMID- 3028655 TI - [Effects of Rehmannia glutinosa, Plastrum testudinis, Aconitum carmichaeli and Cinnamomum cassia on the beta-adrenergic receptors of hyperthyroid rat kidneys]. PMID- 3028656 TI - [Observation on the anti-inflammatory action of propyl gallate]. PMID- 3028657 TI - [Clinical and experimental study on the antitumor effect of cockroach extract]. PMID- 3028658 TI - [Study on the nature of "cold" and "heat" syndromes in traditional Chinese medicine by determination of the amount of catecholamines and cyclic nucleotides excreted in the urine]. PMID- 3028659 TI - [Studies on anti-platelet aggregation effect of Allitridi in hypercholesterolemic rabbits]. PMID- 3028660 TI - Experience with 24-h infusions of ifosfamide/mesna in small cell lung cancer. AB - Two studies were carried out (A and B) in order to assess the effectiveness of ifosfamide administered with mesna (IFO/M) in the treatment of small cell lung cancer. The first study (A) was a cross-over study; the second (B) was a randomized trial, and in B IFO/M was evaluated earlier in the course of the disease. IFO/M given be infusion is effective as second-line therapy and can be administered with other cytotoxics at the doses reported here earlier in the course of the disease. The complete remission rates were high. PMID- 3028661 TI - Ifosfamide by bolus as treatment for advanced non-small cell lung cancer. AB - Ifosfamide at 5 g/m2 was given as a bolus to 48 patients with advanced progressing non-small cell lung cancer. Mesna (5 g/m2) was also given with the ifosfamide, both over 30 min. Further mesna was then given p.o. (3 g/m2) at 4, 8, and 12 h. If oral mesna was not acceptable then one or, if necessary, two 4-h to 6-h infusions (3 g/m2) were administered. A maximum of six courses at 3-weekly intervals was prescribed. A total of 174 courses was administered, and oral mesna was given during 64 courses: discharge was considered possible within 8-10 h after 55% of courses. Haematological toxicity was mild and no renal dysfunction was noted. Two patients became drowsy shortly after ifosfamide, but recovered 24 36 h later. The objective response rate was 29%, with one complete responder. A further 31% of patients (symptomatic responders) with stable disease symptomatically improved after the chemotherapy, by 20 or more points on the Karnofsky scale. The median survival was 5 months for the whole group, and 8 months each for the objective and symptomatic responder groups. Most patients' Karnofsky and respiratory scores improved with the chemotherapy. Ifosfamide with mesna can be given by short infusions, and the mesna given p.o. prevents any urothelial toxicity. Further exploration of short-infusion ifosfamide and mesna therapy would reduce hospitalization and allow for day-case regimens. PMID- 3028662 TI - Experience with ifosfamide combinations (etoposide or DDP) in non-small cell lung cancer. AB - In all, 171 patients with histologically verified non-small cell lung carcinoma were treated with ifosfamide 2.0 g/m2 on days 1-5 in combination with (91 patients) etoposide 120 mg/m2 on day 1. Therapeutic regimens were repeated after 4 weeks. Supportive treatment with mesna (20% of the ifosfamide doses at 0, 4, and 8 h) was performed. Cisplatin treatment was supported by mannitol-induced diuretic hydration. The overall response rate of ifosfamide/etoposide was calculated to be 27%, with 1 complete and 24 partial remissions. The median survival time for all patients was 8.5 months, for responders 14 months (P less than 0.05), for patients with no change 9.5 months, and for patients with tumor progression 4 months. With ifosfamide/cisplatin, there were 4 complete and 21 partial remissions (response rate 35%). The median survival time for all patients was 8.3 months, for responders 11.5 months, and for patients with tumor progression 4 months. Age, sex, and histological tumor type had no significant effect on survival. Patients with better performance stage and limited disease lived significantly longer. The main side-effects of the cisplatin combination were vomiting, bone marrow depression, and neuropathy. The etoposide combination was tolerated better. Urotoxicity was not significant, as a consequence of treatment with mesna. The results show that the combination ifosfamide/etoposide or ifosfamide/cisplatin are effective in the treatment of non-small cell lung cancer, being comparable to other combinations of etoposide/cisplatin and vindesine/cisplatin. Because of the better tolerability, the combination ifosfamide/etoposide is superior to cisplatin combinations. PMID- 3028663 TI - Two randomized trials for alternating polychemotherapy of small cell lung cancer. AB - Patients with small cell carcinoma of the lung (SCCL) were treated in two multicenter trials with different cytostatic drug regimens including ifosfamide. In the first randomized study, including 306 patients, alternating chemotherapy with VP 16, ifosfamide, vindesine (VPIV), adriamycin, cisplatinum, vincristine (APO), and cyclophosphamide, methotrexate, CCNU (CMCC) was compared against standard treatment with ACO (adriamycin, cyclophosphamide, vincristine). It was shown that the alternating therapy resulted in a higher response rate (88% vs 78%) and a longer median survival time (11 months vs 10 months). Regarding toxicity, VPIV was similar to ACO, whereas APO and CMCC had more side-effects, leading to an increase in the number of drop-outs. In the second randomized study 144 patients were treated either with ifosfamide/VP 16 (IVP) or with cisplatinum/VP 16 (PVP). In the case of no further response, no change, or progression the induction therapy was changed to ACO. Interim analyses show that both regimens have similar therapeutic effects; but higher toxicity was observed in patients treated with cis-platinum/VP 16 than in patients treated with ifosfamide/VP 16. According to the response rate in patients treated with ACO after first-line therapy there was less cross-resistance of IVP than of PVP to ACO. PMID- 3028665 TI - Peripheral DTIC neurotoxicity: a case report. PMID- 3028666 TI - Resistance of 7,12-dimethylbenz[a]anthracene-deoxyadenosine adducts in DNA to hydrolysis by snake venom phosphodiesterase. AB - In enzymatic hydrolyses of 7,12-dimethylbenz[a]anthracene (DMBA)-modified DNA isolated from fetal mouse cell cultures, a low concentration of venom phosphodiesterase was sufficient for complete release of DMBA-deoxyguanosine adducts. However, efficient release of DMBA-deoxyadenosine adducts required much higher levels of phosphodiesterase. If these adducts exhibit similarly differential susceptibilities to exonucleases involved in DNA metabolism or repair, each adduct could result in significantly different biological effects in vivo. PMID- 3028664 TI - Potentiation of the chemotherapeutic action of 5'-deoxy-5-fluorouridine in combination with guanosine and related compounds. AB - The effect of inosine, guanosine, and guanosine 5'-monophosphate (GMP) on the antitumor activity of 5'-deoxy-5-fluorouridine (5'-DFUR) was investigated using P388 leukemia and P815 mastocytoma. The antitumor activity of 5'-DFUR was markedly enhanced by coadministration of inosine or guanosine. The increase in lifespan (ILS) of mice treated with 5'-DFUR was augmented by the combination with guanosine or inosine in a dose-dependent fashion, and the maximum ILS was about 160% with the combination, while that in the case of 5'-DFUR alone was only 48% in the P388 leukemia system. The therapeutic ratio (dose at ILSmax/dose at ILS30) of the combination with guanosine or inosine was 333 and 136, respectively, whereas that of 5'-DFUR alone was 3.6. GMP also markedly potentiated the antitumor activity of 5'-DFUR in both P388 leukemia and P815 mastocytoma systems, just as it potentiated the activity of 5-fluorouracil in the latter system. The uric acid level in the serum was elevated after IP injection of guanosine or inosine but the value was much lower in the case of guanosine than in inosine. PMID- 3028667 TI - Characterization and use of polyclonal antibody to Na+,K+-ATPase: immunocytochemical localization in salt glands of the duck. AB - The amount of Na+,K+-ATPase of the avian salt gland increased concomitantly with plasma membrane surface area during salt feeding of ducklings (adaptation), and both enzyme content and membrane surface area decreased upon return to fresh water (deadaptation). In a further study of the enzyme, a marker for plasma membrane biogenesis, polyvalent antibodies were raised to the denatured alpha subunit of the purified ATPase. Antisera did not inhibit enzymatic activity but immunoprecipitated the phosphorylated intermediate of the alpha-subunit. Furthermore, the alpha-subunit, which was not glycosylated, was immunoprecipitated from homogenates of tissue slices metabolically labelled with [35S]-methionine, using antisera raised against either duck salt gland or dog kidney alpha-subunit. The former antisera also recognized the alpha-subunit in the brain, heart, kidney, liver, intestine and skeletal muscle of the duck. Immunocytochemistry with the antisera raised to the duck salt gland alpha-subunit revealed reaction at basolateral as well as apical plasma membrane in the duck salt gland principal cells, with essentially no deposits on peripheral cells, fibroblasts, erythrocytes, endothelial cells and neural elements. Within the principal cells, immunolabelling was also detected on small vesicles, multivesicular bodies and lysosomes; deposits on extracellular debris and vesicles in the lateral and lumenal spaces were also apparent. The labelling patterns were qualitatively but not quantitatively similar in salt glands of control, adapted and deadapted ducklings, and are discussed in the context of a model for plasma membrane biogenesis and turnover in which degradative events may play a major role. PMID- 3028668 TI - Flow cytometric determination of cell cycle phase-specific changes in cellular phosphatase and glycosidase activities. AB - The activities of two phosphatases (E.C. 3.1.3.1 and 3.1.4.1) and four glycosidases (E.C. 3.2.1.21, 3.2.1.30, 3.2.1.31 and 3.2.1.51) were measured by fluorescence spectrophotometry, and flow cytometry, in mitogen-stimulated lymphocytes, and in cultures of Molt-4-F and F-89 cell lines, synchronized by hydroxyurea or thymidine. All enzymes were active throughout the cycle but the activities of three enzymes were elevated at specific points in the cycle, alkaline phosphatase activity increased at G2 + M/G1 boundary and in early S phase, the activity of beta-L fucosidase was elevated in G1 and late S-phase. Orthophosphate diesterase activity was elevated at the G1/S boundary, and during G2 + M. The increase in beta-L fucosidase activity was due to an increased number of cells showing activity, whilst the increase in orthophosphate diesterase activity was attributable to an increase in cellular enzyme activity. Only the activities of orthophosphate diesterase and beta-L fucosidase were measurable by flow cytometry, alkaline phosphatase activity was mainly extracellular, and therefore not detectable by flow cytometric methods employed. PMID- 3028669 TI - Inhibition of lipid peroxidation improves survival rate of endotoxemic rats. AB - The accumulation of lipoperoxide (LPO) is reported to occur in the organs of animals with endotoxemia, where it is accompanied by an activation of xanthine oxidase (XOD) and a depletion of superoxide dismutase (SOD). In the present study, three measures of preventing LPO accumulation, ie, prior treatment with a XOD inhibitor, exogenous supply of enzymatic scavengers, and supplementation with chemical quenchers, were investigated to determine how to improve the survival rate of rats with lethal endotoxemia. Thirty minutes after treatment with various doses of allopurinol, SOD, catalase (CAT), vitamin E (VE), and reduced glutathione (GSH), adult male Wistar rats were subjected to endotoxemia by an intraperitoneal injection of 0.4 mg/100 g of Escherichia coli endotoxin. Allopurinol did not improve survival rates, denoting a lower level of XOD and an almost normal level of SOD in the liver. SOD (9,000 U/100 g) with or without CAT (4,000 U/100 g) markedly increased the survival rate of rats, with complete inhibition of hepatic LPO accumulation and suppression of XOD activity. CAT alone had no salutary effects on survival rate or hepatic LPO. Large amounts of VE (100 mg/100 g) or GSH (50 mg/100 g) slightly suppressed the accumulation of LPO in the liver but had no effect on survival rate. In that exogenous SOD has been considered not to penetrate the cellular membrane because of its high molecular weight, the results suggest that the extracellular spaces are the site of SOD action. Lipid peroxidation of the biomembrane initiated by oxygen free radicals released into extra-cellular space from phagocytes may play an important role in the development of lethality in experimental endotoxemia. PMID- 3028670 TI - Effects of E. coli endotoxin on rat plasma angiotensin converting enzyme activity in vitro and in vivo. AB - Angiotensin converting enzyme (ACE), a glycoprotein, is found in high concentration in pulmonary capillary endothelial cells. Several investigators have studied the relationship between various direct and indirect pulmonary insults and ACE activity and in some cases have found conflict. In an attempt to clarify this relationship we have examined the effect of endotoxin on rat plasma ACE activity in vitro and in vivo. We used the synthetic ACE substrate 3H-BPAP and the assay described by Catravas and Gillis [J Pharmacol Exp Ther 217:263-270, 1981]. In vitro, a statistically significant concentration-dependent reduction in ACE activity was demonstrated (p less than .005). In vivo an intravenous dose of endotoxin (20 mg/kg) alone resulted in no significant change in plasma ACE activity. However, the combination of intravenous endotoxin (20mg/kg) and mild hemorrhage (5-10% of blood volume) caused a statistically significant reduction in plasma ACE activity by 15 min as compared to control rats with hemorrhage only (39% vs. 66%, p less than .005). This reduction persisted at 30 and 60 min. However, by 180 min ACE activity was no longer statistically different from control values. We have demonstrated an acute reduction of plasma ACE activity in the endotoxemic rat that appears to be dependent on the amount of circulating endotoxin and the presence of mild blood loss. PMID- 3028671 TI - PGH synthase and lipoxygenase generate superoxide in the presence of NADH or NADPH. AB - Purified PGH synthase when acting on arachidonic acid in the presence of reduced nicotinamide-adenine dinucleotide or reduced nicotinamide-adenine dinucleotide 3' phosphate generated superoxide in burst-like fashion. In eight experiments using different batches of enzyme, the mean +/- SE rate of superoxide generation from 100 U of enzyme measured as the superoxide dismutase-inhibitable reduction of cytochrome c was 5.06 +/- 0.19 nmol/min in the first minute and 0.35 +/- 0.03 nmol/min subsequently. Optimum rates of superoxide were seen at concentrations of reduced nicotinamide-adenine dinucleotide in excess of 80 microM and reduced nicotinamide-adenine dinucleotide 3'-phosphate in excess of 100 microM. Using prostaglandin G2 or linoleic acid as substrate rather than arachidonate also resulted in superoxide generation. When prostaglandin H2 was used as substrate, no superoxide was generated. The rate of superoxide generation was markedly inhibited by cyclooxygenase inhibitors. Superoxide generation was also observed during the action of lipoxygenase on either linoleic or arachidonic acid in the presence of reduced nicotinamide-adenine dinucleotide or reduced nicotinamide adenine dinucleotide 3'-phosphate but not in their absence. Indomethacin had no effect on superoxide generation from lipoxygenase. We conclude that PGH synthase and lipoxygenase produce superoxide via a side-chain reaction dependent on the presence of suitable reducing cosubstrate. This mechanism is analogous to that described for peroxidases in general. PMID- 3028672 TI - The effects of histamine on contraction frequency, sodium influx, and cyclic AMP in cultured rat heart cells. AB - Histamine has been shown to have both positive inotropic and chronotropic effects. To evaluate the chronotropic effects, spontaneously contracting monolayers of cultured rat myocardial cells were treated with histamine, 10(-7) M 10(-4) M. This resulted in a dose-dependent increase in contraction frequency reaching a maximum in 10(-5) M histamine. Contraction frequency (mean +/- SEM) increased from a control of 121 +/- 5 contractions per minute to 153 +/- 4.5, 181 +/- 9, 212 +/- 4, and 216 +/- 1 in 10(-7) M, 10(-6) M, 10(-5) M, and 10(-4) M histamine, respectively (for each n = 10, p less than 0.001). The effect was time dependent, taking 30 minutes to develop fully. Changes in contraction frequency were accompanied by parallel dose- and time-dependent increases in the verapamil sensitive sodium influx. Verapamil-sensitive sodium influx (pmol/cm2/sec) increased from a control of 10.45 +/- 1.44 (mean +/- SEM) to 24.34 +/- 2.41 and 32.57 +/- 2.35 at 10- and 30-minute treatment with 10(-6) M histamine (n = 5, p less than 0.001). These data fit the previously described relation between verapamil-sensitive sodium influx and contraction frequency in these cells. Cimetidine (10(-4) M) but not diphenhydramine (10(-4) M) abolished both the contraction frequency and sodium influx response to histamine. Subsequent studies showed a dose- and time-dependent elevation of cyclic adenosine monophosphate (cAMP) with histamine treatment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3028673 TI - Clinical experimental considerations on the presence of set uterine pacemakers. AB - The authors studied the physiology of human uterine muscle contraction. Clinical experimental results obtained from elementary electrophysiological, electrohysterographic and pharmacological studies using PGs suggested that uterine muscle contraction does not originate in "pacemaker" areas. These data also showed that the electric phenomenon is not propagated along preferential "rails". PMID- 3028674 TI - Neutrophil dysfunction. Case studies and review. PMID- 3028675 TI - Urinary cyclic adenosine monophosphate discriminates subsets of patients with osteoporosis. PMID- 3028676 TI - Determination of serum neuron-specific enolase by differential immunocapture. AB - A new method for the determination of serum neuron-specific enolase is presented. It consists of two steps: first, an immunocapture of gamma-subunit containing isoenzymes by absorption on immobilized anti-gamma antibodies; second, bioluminescence assay of enolase activities in untreated control samples and in the supernates of antibody treated samples. Total and alpha alpha activities are obtained, from which the neuron-specific enolase activity (alpha gamma + gamma gamma) can then be calculated by difference. As compared to the procedures currently in use, the immunocapture method is very rapid (30 min) and is more suitable for small series of determinations as needed in clinical chemistry applications. Reference interval values for serum found by this method agree with published data. When tested with samples from patients suffering from neuroblastoma or small cell lung cancer, it confirms the specific elevations in neuron-specific enolase activity previously described for these cancers, using other analytical approaches. PMID- 3028677 TI - Estimation of oestrogen and progesterone receptors in chronic rhinitis. AB - The turbinates of 38 patients with chronic rhinitis were examined biochemically for oestrogen and progesterone receptors. Low levels of oestrogen-receptor-like activity (1-20 fmol/mg protein) were found in 50% of patients of both sexes. Progesterone receptor activity was also weak (1-16 fmol/mg protein) but was present only in 5 female patients. Immunocytochemical assay failed to demonstrate focal areas of oestrogen receptor activity. One juvenile nasopharyngeal angiofibroma was negative for both oestrogen and androgen receptors. Other possible mechanisms of hormonal action are considered. PMID- 3028678 TI - Juvenile angiofibroma: imaging by magnetic resonance, CT and conventional techniques. AB - Thirty patients with histologically verified angiofibromata have been investigated over a period of 14 years. They have been examined by conventional radiographic techniques and computerized tomography, and more recently 4 patients have been scanned by magnetic resonance. CT studies of patients with small tumours have shown that the point of origin is at the sphenopalatine foramen. The tumour enlarges the foramen and erodes bone locally giving rise to characteristic signs both on plain X-ray and on CT scan. The value of magnetic resonance imaging is assessed and it is concluded that in the presence of the characteristic 'antral sign' on plain X-ray, 3-plane magnetic resonance is now the method of choice to show the extent of the tumour pre-operatively. Magnetic resonance can also show the vascular nature of the angiofibroma by the demonstration of large vessels, shown as dark areas of negative signal within the tumour mass. With this new method of investigation available, angiography should now only be performed if embolization is deemed necessary prior to surgical removal of the angiofibroma. PMID- 3028679 TI - Adenoid cystic carcinoma. PMID- 3028680 TI - Primary biliary cirrhosis: assessment of the quantitative importance of the adenine nucleotide translocator protein as a major mitochondrial antigen. AB - The adenine nucleotide translocator protein (ANT) is the first well-characterized mitochondrial polypeptide to be identified as an antigen for antimitochondrial autoantibodies (AMA) in PBC sera. Because of the potential use for a highly purified antigen as a tool in studying the aetiology of PBC, we have undertaken an assessment of the quantitative importance of ANT as a PBC-specific mitochondrial antigen. Immunoblotting and ELISA techniques were used. Both methods reveal PBC antibodies against isolated rat liver ANT. However, competitive ELISA experiments using purified rat liver ANT as the competing antigen show that anti-ANT antibodies in PBC serum comprise only a fraction of the total AMA. Furthermore, both ELISA and immunoblotting experiments show that rat liver ANT is not a specific antigen for PBC autoantibodies. Sera from patients with SLE, chronic active hepatitis, and sera from normal, control patients, have nearly the same, or higher, ANT antibody titres. Thus, ANT is not a good candidate as an antigen for the diagnosis of PBC. PMID- 3028681 TI - Postjunctional vascular smooth muscle alpha-2 adrenoreceptors in human autonomic failure. AB - Animal studies have suggested that vasoconstrictor alpha-2 adrenoreceptors exist on vascular smooth muscle cells. We tested this hypothesis in a patient with severe autonomic failure who demonstrated a pressor response to oral clonidine (a selective alpha-2 adrenoreceptor partial agonist). After clonidine 0.8 mg orally, mean arterial pressure rose by 54 mm Hg. After pretreatment with prazosin (a selective alpha-1 adrenoreceptor antagonist) and confirmation of alpha-1 blockade, clonidine 0.8 mg still raised mean arterial pressure by 43 mm Hg. After pretreatment with yohimbine (a selective alpha-2 adrenoreceptor antagonist), clonidine 0.8 mg elevated mean arterial pressure by 13 mm Hg. Since alpha-1 antagonism does not block, and alpha-2 antagonism does not block the pressor effect of clonidine, we conclude that clonidine raised blood pressure in this severely affected autonomic failure patient by vascular alpha-2 adrenoreceptor stimulation. Thus, this provides pharmacological evidence that postjunctional vascular smooth muscle alpha-2 adrenoreceptors exist in man and can modulate blood pressure. PMID- 3028682 TI - Effects of differing purified cellulose, pectin and hemicellulose fiber diets on mucosal morphology in the rat small and large intestine. AB - The effects of increasing amounts of differing single-fiber sources in chemically defined and nutritionally-equivalent diets on morphometric measurements of the rat small and large intestinal mucosa were examined. Male Wistar rats received a fiber-free diet, or diets containing 4.5% or 9.0% cellulose, pectin, or hemicellulose. Cellulose and pectin increased jejunal (but not ileal) villus and crypt heights, whereas hemicellulose increased ileal (but not jejunal) mucosal height. Moreover, cellulose and pectin (but not hemicellulose) increased crypt heights in cecum and proximal (but not distal) colon. Parallel increases in the mitotic index were seen with cellulose and pectin diets, but there was a decrease with hemicellulose, suggesting different mechanisms for altered patterns of cell renewal induced by orally-administered single-fiber sources. These changes indicate that differing single-fiber sources exert differential structural effects along the length of the small and large intestine in the rat. PMID- 3028683 TI - Familial polyposis coli in childhood. AB - Familial polyposis coli has generally been considered a disorder of adulthood that is rarely identified in children. We report 3 affected kindreds in which the disorder was diagnosed in 6 of 11 potentially affected children between 8 and 16 years of age. Experience with these kindreds demonstrates that polyps frequently develop during childhood in affected individuals. Colonoscopy was found to be preferable to the air-contrast barium enema, in that it was as sensitive a technique for detection of adenomas, and in that it also permitted collection of biopsies for histologic confirmation. We recommend that colonoscopy be performed late in the first decade of life before symptoms develop. Total colectomy, rectal mucosectomy, and ileo-anal anastomosis eliminated the risk of malignancy, preserved anal sphincter function by both clinical and manometric assessment, and was readily adapted to by these children. PMID- 3028684 TI - Primary intracranial choriocarcinoma: a case report. AB - A 10-year-old girl had a primary choriocarcinoma of the posterior third ventricle. Craniotomy a few hours before death did not yield any tumor tissue. At autopsy, an extensively hemorrhagic tumor abutted the pineal gland. Immunostains were positive for beta-human chorionic gonadotropin (beta-HCG) but were negative for alpha-fetoprotein and carcinoembryonic antigen. The presence of beta-HCG in serum or cerebrospinal fluid may be used as a diagnostic marker and monitor of therapy. HCG is, however, not a unique marker for trophoblastic neoplasms, as a significant number of intracranial germinomas contain cells that are beta-HCG positive. Because of the rarity of primary extragenital choriocarcinomas and the much more common occurrence of metastases of genital choriocarcinomas, it is doubtful whether any investigation less than detailed autopsy can prove the extragenital origin of the tumor. PMID- 3028685 TI - An autonomous functioning cystic thyroid adenoma. An unusual occurrence. AB - An asymptomatic thyroid nodule that appeared cystic on ultrasound was seen in a 14-year-old girl. Tc-99m pertechnetate thyroid imaging was expected to show a cold nodule, but surprisingly revealed a hyperfunctioning nodule. A partial thyroidectomy revealed an hemorrhagic cystic adenoma. The occurrence of a cystic nodule, which continued to show hyperfunction, is unusual. PMID- 3028686 TI - Lymphoscintigraphic studies of lymphatic drainage from the testes. AB - Two colloidal radiopharmaceuticals, Au-198 and Tc-99m antimony, were used to evaluate the lymphatic drainage of the testis in experimental animals and humans. One to 24 hours after direct intratesticular injection of Au-198 colloid in dogs and 4-6 hours after injection of Tc-99m antimony colloid in men, distribution within retroperitoneal lymph nodes was demonstrated. Uptake within the para aortic lymph nodes primarily draining the testis was decreased following proximal ligation of the spermatic vessels in dogs. Testicular lymphoscintigraphy successfully demonstrated an intact spermatic cord lymphatic communication to the para-aortic nodes in five of six patients with chronic lower-extremity lymphedema. When the intact testicle and spermatic cord were transposed to the thigh in a patient with chronic lymphedema of the lower extremity, percutaneous pedal lymphoscintigraphy successfully demonstrated uptake within the para-aortic lymph nodes draining the ipsilateral testis. PMID- 3028687 TI - Radionuclide findings in arteriovenous fistula between left pulmonary artery and vein. AB - Right-to-left extra cardiac shunts are usually congenital and are rarely due to trauma or complicating thoracic or vascular surgery. A case of a right-to-left cardiac shunt due to an arteriovenous fistula between the left pulmonary artery and the left pulmonary vein is reported. This may be the first report of this abnormality. The etiology is not clear. The case was investigated only by radionuclide procedures because the general condition of the patient was so poor that invasive procedures could not be undertaken. PMID- 3028688 TI - Hepatoblastoma. AB - Hepatoblastoma is a rare hepatic tumor generally presenting during the first three years of life as an enlarging abdominal mass. Other symptoms are nonspecific; however, it may be associated with hemihypertrophy, virilization, and osteoporosis. The serum bilirubin infrequently is elevated, but up to 2/3's will have elevated serum alpha fetoprotein as a tumor marker. The overall survival rate is 35% in survivors who underwent a complete resection. PMID- 3028689 TI - Bone and gallium scan findings in malignant fibrous histiocytoma. Case report with radiographic and pathologic correlation. AB - Malignant fibrous histiocytoma (MFH) is the most common soft tissue malignancy in adults. The Ga-67 citrate scan findings of an extremity-located MFH, the most common location of this neoplasm, have never been published in English language journals to the best of the authors' knowledge. Ga-67 citrate and Tc-99m MDP scans of the thigh mass accurately depicted the tumor's local extent, including the presence of central ischemic necrosis within the tumor, and the absence of adjacent osseous involvement and distant metastases, as correlated with computed tomography, angiography, and pathologic examinations. PMID- 3028690 TI - Imaging studies of patients with malignant fibrous histiocytoma using C-11-alpha aminoisobutyric acid (AIB). AB - Alpha-aminoisobutyric acid (AIB), a synthetic, nonmetabolized amino acid which is rapidly transported into viable cells by the A-type or alanine-preferring amino acid transport system, has been labeled with the short-lived, positron-emitting radionuclide carbon-11. Carbon-11 labeled AIB is currently being evaluated as a tumor imaging agent for in vivo amino acid transport studies in patients with cancer. In this study, C-11 AIB was used to image two patients with malignant fibrous histiocytoma (MFH), a pleomorphic sarcoma. Following intravenous administration of C-11 AIB, tumors in the distal femur of one patient and in the anterior chest wall of another patient were well visualized using high energy gamma scintigraphy. Since therapy may alter the accumulation of amino acids in tumor tissue, studies using C-11 AIB in patients with MFH before and after chemotherapy are in progress. PMID- 3028691 TI - Hyperthyroidism and the single lobe. AB - A practical approach to the hyperthyroid patient with a single lobe visualized on thyroid scintigraphy, and its impact on therapy is discussed. An illustrative case is also presented. PMID- 3028692 TI - Localization of abnormal parathyroid gland(s) using thallium-201/iodine-123 subtraction scintigraphy in patients with primary hyperparathyroidism. AB - Tl-201/I-123 subtraction scintigraphy was performed in 17 patients with clinical symptoms and biochemical measurements suggestive of primary hyperparathyroidism. Nineteen abnormal sites were identified. These results were correlated with PTH measurements and surgical findings. Three sites were considered unrelated to the parathyroid glands, two corresponding to palpable thyroid nodules and one to muscle uptake of unknown origin. One scintigram did not reveal either of two abnormal glands while two others were considered falsely positive in view of surgical failure. Fourteen sites corresponded to abnormal parathyroid gland at surgery; five glands, weighing more than 2000 mg, could be correctly located on the Tl-201 scintigraphy prior to the subtraction procedure; six glands, weighing between 500 and 2000 mg, were easily localized after the subtraction procedure; three glands, weighing between 180 and 200 mg, were correctly localized after further manipulation of the subtraction procedure. In a patient with parathyroid hyperplasia, one gland, weighing 150 mg, was not located and another was not found upon surgery. Overall sensitivity was 87.5%. A positive correlation between PTH levels, tumor weight, and ease of detection on scintigraphy was found. This correlation was particularly useful in excluding large abnormal uptake related to thyroid disorder or artifact. The results suggest that Tl-201/I-123 parathyroid scintigraphy could become an alternative to Tl-201/Tc-99m parathyroid scintigraphy, with possibly improved detection of low weight abnormal parathyroid glands. PMID- 3028693 TI - Sodium transport across erythrocyte membranes in diabetes mellitus. AB - The activity of the erythrocyte ouabain sensitive Na/K ATPase pump was studied in 20 Type 1 diabetics, 15 Type 2 diabetics and 20 healthy controls. All diabetics were normotensive, caucasian and non-obese diabetics. The diabetics were assessed for retinopathy and had HbA1 estimated. The erythrocyte sodium concentration, membrane permeability to sodium and ouabain-sensitive sodium efflux rate constant (ERCos) were determined. The Type 1 diabetics had a significantly reduced ERC, mean +/- SD 0.198 +/- 0.028 hr-1 vs control 0.264 +/- 0.054 hr-1 (p less than 0.001). A similar reduction in ERCos was seen in the Type 2 diabetics when compared to controls (0.229 +/- 0.059 hr-1 p less than 0.005). There was also a significant difference between the 2 groups of diabetics (p less than 0.05). There was no significant difference in intracellular sodium concentrations. Membrane permeability was reduced in the Type 1 diabetics (3.69 +/- 0.79 mmol/kg dry wt/hr) compared to the control group (4.56 +/- 0.79 mmol/kg dry wt/hr, p less than 0.01). Diabetic control (HbA1) was positively correlated with ERCos, but no significant differences were seen in the ERCos between complicated and uncomplicated Type 1 diabetics. In vitro studies of normal erythrocytes did not show an effect on pump activity of increasing glucose or insulin concentration during a preincubation study. These results demonstrate that there is a reduction in activity of the ouabain-sensitive Na/K ATPase pump and a reduction in membrane permeability on the diabetic erythrocyte which is most marked in Type 1 diabetics. PMID- 3028695 TI - [Phantom substances in the quantitative evaluation of magnetic resonance T images. II. Effect of relaxation times on magnetic resonance T-intensity values]. AB - Reference material for quantitative MR Imaging (paramagnetic agar gel) is examined with regard to effects of small T1 and T2 variations on MRI signals. For some pulse sequences that are important in clinical practice, it can be shown that SE images are very sensitive to T2 variations, whereas IR images are influenced by both T1 and T2. PMID- 3028694 TI - Neglected radiologic signs of the glucagonoma syndrome. AB - A case of glucagonoma where repeated gastrointestinal examinations revealed excessive mucosal fold thickening of the duodenum and small bowel with a markedly delayed transit time is reported. These findings in the appropriate clinical setting led us to persevere with further investigations despite equivocal ultrasound and CT examinations. We wish to emphasize the importance of the classical gastrointestinal findings in the diagnosis of the glucagonoma syndrome. PMID- 3028697 TI - Multilocular cystic nephroma: report of three cases. AB - Multilocular cystic nephroma is an uncommon renal neoplasm which in childhood usually presents as an abdominal mass. The same condition is also described as multilocular kidney and multilocular cystic hamartoma. The clinical presentation, and radiological findings in three cases of multilocular cystic nephroma are described. PMID- 3028696 TI - [Plain imaging of meningioma in NMR tomography--retrospective contrast optimization by image synthesis]. AB - Synthetic images of the tumour region were computed in the case of ten meningiomas for the purpose of determining retrospectively pulse sequences and sequence parameters supplying an optimal contrast between tumour and adjacent tissue in plain imaging. It was found that best results were obtained via the partial saturation/spin-echo sequence with a long recovery time (TR approximately greater than 2,000 msec) and a long maximum echo delay (TE max approximately greater than 200 msec). A comparable contrast quality can be attained by using the inversion-recovery sequence with an inversion time T1 approximately 450 msec. Nevertheless, it was not possible in our study to identify two small meningiomas with a diameter of just below one centimetre by studying the MR tomogram. PMID- 3028698 TI - Ion flux and Na+,K+-ATPase activity of erythrocytes and leucocytes in thyroid disease. AB - In hyperthyroidism, erythrocytes show decreased Na+,K+-ATPase activity, decreased [3H]ouabain binding capacity (an index of the number of sodium pumps) and decreased active sodium and potassium flux rates, with a high intracellular sodium concentration. As erythrocytes are non-nucleated and atypical cells, we have studied electrolyte status in thyroid disease using mixed leucocytes as well; the results obtained differed from those in erythrocytes. When compared with findings in healthy subjects, leucocyte Na+,K+-ATPase activity, [3H]-ouabain binding capacity, total and active rubidium (used instead of potassium) influx were all significantly increased in untreated hyperthyroidism and decreased in untreated hypothyroidism. In hyperthyroidism, there was also a decrease in plasma potassium, an increase in sodium efflux rate and efflux rate constant, but no significant changes in cell sodium and potassium concentrations. All these changes returned to normal in successfully treated patients. There was a significant correlation between these abnormalities of electrolyte status and thyroid disease status (as serum thyroid stimulating hormone and free thyroxine). PMID- 3028699 TI - A randomized controlled trial of the effect of dietary fibre on blood pressure. AB - Eighty-eight healthy omnivores with normal blood pressure participated in a randomized, controlled, cross-over trial of the effect on blood pressure of increasing dietary fibre intake. Subjects were randomly allocated to a control group eating a low fibre diet throughout, or to one of two experimental groups eating a high fibre diet for one of two 6-week experimental periods. Changes in body weight, other dietary constituents and lifestyle factors were avoided as far as possible. Twenty-four hour diet records showed a substantial increase in dietary fibre when subjects were on the high fibre diet. There was no consistent effect of change in dietary fibre intake on group mean systoloic or diastolic blood pressures. Adjusting blood pressures for changes in other dietary components, plasma lipids, electrolytes, body weight and other lifestyle variables did not alter this result. These findings do not support the hypothesis that the blood pressure lowering effect of a vegetarian diet is solely due to an increase in fibre intake. PMID- 3028701 TI - Lack of angiotensin I accumulation after converting enzyme blockade by enalapril or lisinopril in man. AB - In nine normal volunteers, a series of five venous blood samples was obtained before and up to 24 h after converting enzyme inhibition by a single oral dose of enalapril or lisinopril. Plasma renin activity and blood angiotensin I were measured. A close linear relationship was found between the increase in plasma renin activity and the increase in blood angiotensin I. The linear correlation between plasma renin activity and blood angiotensin I remained after converting enzyme inhibition. Thus, the rise in angiotensin I after inhibition of the conversion of angiotensin I to angiotensin II is due to an enhanced release of renin rather than to accumulation of angiotensin I. PMID- 3028700 TI - Reduced brain Na+, K+-ATPase activity in rats with galactosamine-induced hepatic failure: relationship to encephalopathy and cerebral oedema. AB - Previously we have shown that sera from patients with fulminant hepatic failure (FHF) will inhibit partially purified rat brain Na+, K+-ATPase and sodium efflux from human leucocytes in vitro. Similar inhibition may be involved in the pathogenesis of encephalopathy and cerebral oedema in these patients. In the present study we have attempted to establish whether the activity of brain Na+, K+-ATPase is decreased in vivo in rats with D-galactosamine induced hepatic failure using homogenates of snap-frozen brains. Na+, K+-ATPase activity was significantly reduced in the forebrain region at the stage of mild encephalopathy (43 h after injection), while at the deeper stage of coma (43-53 h after injection) enzyme activity was further reduced in the forebrain region and was also significantly reduced in the hindbrain region. Ouabain insensitive ATPase activity was not significantly altered at any time. While a significant increase in the water content (0.5%) of the hindbrain region was found 43 h after galactosamine, there was no clear correlation between the development of cerebral oedema and the reduction of Na+, K+-ATPase activity. The activity of partially purified normal rat brain Na+, K+-ATPase was 15% lower when incubated with sera from rats in the deep stage of coma compared with control sera. These data support other evidence that the reduction in brain Na+, K+-ATPase is likely to be due to toxic substance circulating in serum which have been shown to inhibit this enzyme in vitro and to cause coma when administered to normal animals. PMID- 3028702 TI - Anaphylactic reaction to intravenous conjugated estrogens. PMID- 3028703 TI - Hypocapnia and hypercapnia in the dog: effects on myocardial blood-flow and haemodynamics during beta- and combined alpha- and beta-adrenoceptor blockade. AB - We have recently demonstrated an increase in myocardial blood-flow (MBF) during systemic hypercapnia independent of myocardial oxygen consumption. During hypocapnia no significant decrease of MBF was observed; however, a significant increase of myocardial oxygen extraction resulted. The present study was undertaken to examine whether these effects were caused by changes in the sympathoadrenal activity. During beta-adrenoceptor blockade and combined alpha- and beta-blockade the effects of hypocapnia and hypercapnia on MBF and haemodynamics were examined. Closed-chest dogs were anaesthetized with pentobarbital and hypocapnia was induced by hyperventilation. Carbon dioxide was added to the inspiratory gas to create normocapnia and hypercapnia. Hypocapnia did not result in any changes of MBF, coronary vascular resistance or myocardial oxygen extraction, neither during beta-adrenoceptor blockade nor during combined alpha- and beta-blockade. Hypercapnia did not increase MBF, neither during beta blockade nor during combined alpha- and beta-blockade. However, both myocardial oxygen consumption and mechanical performance of the heart were reduced during hypercapnia. Thus, a relative myocardial overperfusion, indicated by decreased coronary vascular resistance and myocardial oxygen extraction, was observed during hypercapnia. In conclusion, the unaltered MBF during hypocapnia and the coronary overperfusion during hypercapnia were not related to changes in the sympathoadrenal activity. PMID- 3028704 TI - Comparison of Isolab Quik-Sep and Quik-Sep System II for separation of immunoglobulins G and M. AB - Use of Quik-Sep System II (QSII) ion-exchange chromatography increased the detection of CMV-directed immunoglobulin M (IgM) compared to the prototype Quik Sep System (QSI) (27 of 87 [31%] IgM-positive versus 20 of 87 [24%]). However, an increase in the number of false-positive rheumatoid factor-containing sera was found (15/52 [29%] versus 8/52 [15%]). Total immunoglobulin G (IgG) and IgM concentrations were higher in the QSII eluates compared to those from QSI, although total IgM recovery did not exceed 36% in either system. PMID- 3028706 TI - Comparison of MRC-5 and HFF cells for the identification of cytomegalovirus in centrifugation culture. AB - Detection of cytomegalovirus (CMV) by centrifugation culture was compared in MRC 5 and HFF cells. At 24 hr 47 of 130 specimens (36%) were positive for CMV, 45 in MRC-5, and 35 in HFF cells. At 48 hr 45 specimens were positive in both MRC-5 and HFF cells. MRC-5 cells appear to be superior for the detection of CMV at 24 hr in centrifugation culture; however, both lines are comparable at 48 hr. PMID- 3028705 TI - Comparison of the Ortho Cultureset (Vero cells) and Difco Cellmatics system (mink lung and primary rabbit kidney cells) for the detection of herpes simplex virus from clinical specimens. AB - Two Cellmatics (Difco) Herpes simplex virus (HSV) detection systems, one with mink lung cells (ML) and the other with primary rabbit kidney cells (PRK) and the Cultureset (Ortho) system with Vero cells were compared for their ability to detect previously positive specimens. One hundred fifty patients specimens positive for HSV in either Vero or PRK cells prior to freezing at -70 degrees C were thawed once, diluted to 1:8, and inoculated in duplicate into each cell system. Positive cultures were detected using each kit's immunoperoxidase method. All three cell lines were stained at 24 hr followed by a final staining of negative cultures at 48 hr for Cultureset and at 72 hr for Cellmatics as per kit instructions. At 24 hr percent detection was: Cellmatics (ML) = 85%, Cellmatics (PRK) = 81%, Cultureset (Vero) = 74%. At final staining percent detection was 94%, 92%, and 84%, respectively. The Cellmatics system has the advantage of fewer steps in the staining procedure and no manipulation of coverslips because staining is performed in the tubes. As employed in these commercial systems, the Cellmatics system with ML cells is more sensitive than the Cultureset with Vero cells (p less than 0.001) and comparable to the Cellmatics system with PRK. PMID- 3028707 TI - Increased prolylhydroxylase activity in chondrocytes but not synovial cells from adjuvant arthritic rat knees. AB - Prolylhydroxylase and galactosylhydroxylsyl glucosyl (GGH) transferase were assayed in cells cultured from cartilage and synovium (adipose type) removed from normal and adjuvant arthritic rat knees. There was a significant increase in prolylhydroxylase in chondrocytes from arthritic knees compared to the normals but no change in GGH transferase activity was found. In the cells derived from the synovium no variation in either enzyme was observed. No significant changes in DNA or protein content was found in either type of cells from the arthritic joints. Comparison of the prolylhydroxylase activity in the supernatant and pellet fractions prepared from chondrocytes from normal and arthritic joints, demonstrated that the majority of the activity was recovered in the supernatant fraction rather than the pellet in arthritic chondrocytes. The data presented indicate that the degree of hydroxylation of cartilage collagen alters in arthritis. PMID- 3028708 TI - Suppression of matrix protein synthesis by herpes simplex virus type 1 in human endothelial cells. AB - The synthesis of matrix proteins was investigated in cultures of human umbilical vein endothelial cells (EC) infected with herpes simplex virus type 1 (HSV-1). EC cultures were either mock-infected or infected for 1 hour at a multiplicity of infection (MOI) of 5 or 20 infectious particles per cell. Synthesis was followed by determining [14C]-proline or [35S]-methionine incorporation into non dialyzable proteins. Using SDS-PAGE we observed that synthesis of fibronectin (FN), type IV procollagen and thrombospondin (TSP) was inhibited in infected cultures as early as 2 hours becoming almost complete by 15 hours post-infection. The degree of inhibition of matrix protein synthesis was dependent on the dose of the virus inoculum (MOI 20 greater than MOI 5) and varied with the particular matrix protein, i.e. shut-off of type IV collagen occurred first followed by that of FN and then TSP. Pulse-chase experiments suggest that the absence of labeled matrix protein in the medium of infected cultures is not due to accumulation of protein within the infected cells since there was an equal and parallel reduction in the cell layer. Suppression of matrix proteins in infected cultures was confirmed by measuring the level of FN and TSP in uninfected and infected cultures in an enzyme-linked immunosorbent assay (ELISA). The three proteins were identified by ELISA, electroimmunoblot, immunoprecipitation and ion-exchange chromatography. The data suggest that HSV-1 infection of human EC suppresses matrix protein synthesis; the degree and time of complete shut-off varies with the protein and the virus dose. PMID- 3028710 TI - Collagen biosynthesis and degradation during deposit and resorptive phases of carrageenin granuloma. AB - Collagen metabolism was studied in both, the developing and resorptive phases of carrageenin granuloma. The rate of collagen biosynthesis relative to the rate of total protein synthesis is almost twice as high during the deposit phase compared with the resorptive phase of the same process. Total collagenolytic activity appears to be approximately the same in both, deposit and resorption stages of the granuloma. Notwithstanding, the enzyme is fully active during resorption; whereas half of this activity is in a latent form during deposit. Collagenolytic activity increased remarkably when assayed at 30 mM CaCl2 as compared to 10 or 50 mM. PMID- 3028709 TI - Characterization of a specific antiserum to rabbit stromelysin and demonstration of the synthesis of collagenase and stromelysin by stimulated rabbit articular chondrocytes. AB - An antiserum to rabbit bone stromelysin (proteoglycanase) was raised in sheep and characterized as specific, recognizing the enzyme from both different tissue sources and different species. This antiserum and a specific antiserum to rabbit bone collagenase were used in the study of metalloproteinase production by rabbit articular chondrocytes stimulated with either interleukin 1 or mononuclear cell conditioned medium. It was shown by electroimmunoblotting that the apparently co ordinate (mole:mole) induction of collagenase and stromelysin activity with time correlated in either case with an increase in enzyme protein. The stimulated production of both enzymes could be modified in parallel by a variety of compounds. Immunohistochemical studies confirmed that although most cells were producing both metalloproteinases simultaneously, some chondrocytes produced detectable levels of only one. The data are discussed in relation to the mechanisms of breakdown in connective tissues. PMID- 3028711 TI - Functional characterization of the enzymes with 2,3-bisphosphoglycerate phosphatase activity from pig skeletal muscle. AB - In pig skeletal muscle exist four enzymes with 2,3-bisphosphoglycerate phosphatase activity. Two of them (forms I-A and I-C) are multi-functional enzymes which, in addition to the phosphatase activity, possess 2,3 bisphosphoglycerate synthase and phosphoglycerate mutase activities. The other two enzyme forms (II-A and II-B) only show the phosphatase activity. The four enzymes differ in substrate specificity. Form I-C is highly specific for glycerate 2,3-P2; form I-A also hydrolyzes the monophosphoglycerates and forms II A and II-B are specific for phosphoester bonds adjacent to a C-1 carboxylic group. The enzymes possess similar Km, Kcat and optimum pH value, but they are differently inhibited by the reaction products. They are also differently affected by glycolate-2-P (their main activator) and by other modifiers. Probably form I-A, which corresponds to M-type phosphoglycerate mutase, is the main enzyme implicated in the breakdown of glycerate 2,3-P2 in pig muscle. PMID- 3028712 TI - Computed tomography of the parapharyngeal space. AB - The parapharyngeal space is a potential anatomic space of appreciable clinical importance which, because of its location deep within the neck, has historically been difficult to evaluate by physical examination and conventional imaging methods. Computed tomography has provided an important and accurate means of noninvasively defining both the anatomy and pathology of this area. This discussion will initially pertain to the normal anatomy and anatomical relationships of the parapharyngeal space. Traditional methods of imaging will be briefly reviewed but emphasis will be on CT and the most appropriate means of utilization of CT. The more important pathology will be included in order to illustrate the advantages of current imaging techniques. PMID- 3028713 TI - Cross-infection enhancement among African flaviviruses by immune mouse ascitic fluids. AB - Cross-infection enhancement of seven African flaviviruses by subneutralising concentrations of antibody in immune ascitic fluids was investigated in P388D1 cell culture. Infection by all the seven flaviviruses tested was enhanced by homologous and at least one of six heterologous immune mouse ascitic fluids (IMAF) tested. Enhancement ratios and enhancing antibody titres were higher in homologous than in heterologous enhancement. Zika, Wesselsbron, Uganda S and West Nile viruses were enhanced in culture by all the IMAF tested. Enhancement of Dakar bat and Yellow fever viruses was produced by five heterologous IMAF, but Potiskum virus was enhanced by one heterologous flavivirus antibody. The antibody to Potiskum virus was the most potent mediator of heterologous infection enhancement; all six heterologous flaviviruses were markedly enhanced by this antibody. PMID- 3028714 TI - Mapping of the gene for anti-mullerian hormone to the short arm of human chromosome 19. AB - The gene coding for human anti-Mullerian hormone (AMH) was localized to subbands p13.2----p13.3 on chromosome 19, using in situ hybridization and Southern blot analysis of a panel of man-mouse and man-hamster somatic cell hybrids. PMID- 3028715 TI - The AluI-induced bands in great apes and man: implication for heterochromatin characterization and satellite DNA distribution. AB - Restriction endonucleases have recently been proved to be active on fixed chromatin, producing differences in staining of metaphase chromosomes. In this paper we show the results obtained by treating the metaphase chromosomes of Pan troglodytes, Pan paniscus, and Gorilla gorilla with the restriction enzyme AluI. These results demonstrate qualitative differences in the telomeric heterochromatin between Pan and Gorilla despite the fact that these areas appear homogeneous in the two genera by the C-banding method. The results found with individual chromosomes in the different species also appear relevant, in the light of the evolutionary relationships between these nonhuman primates and man. Lastly, the results suggest the presence, in great apes, of some highly repetitive DNA sequences different from the human satellites I-IV. PMID- 3028716 TI - Integration sites of human papillomavirus 18 DNA sequences on HeLa cell chromosomes. AB - Human papillomaviruses (HPV) 16 and 18 are closely linked with human genital cancer. In most cervical carcinomas, viral sequences are integrated into the host genome. HeLa, a cervical carcinoma cell line, has multiple copies of integrated HPV 18 DNA. In this study, in situ chromosome hybridization was used to assign the integration sites of HPV 18 DNA sequences on HeLa cell chromosomes. Four sites of hybridization were identified at 8q23----q24, 9q31----q34, p11----p13 on an abnormal chromosome 5, and q12----q13 on an abnormal 22. Three of these sites correspond with the locations of MYC, ABL, and SIS protooncogenes, and are at or in close proximity to fragile sites. The chromosomal localization of HPV 18 DNA may be useful in assessing the role of viral integration in the development of this malignancy. PMID- 3028717 TI - Hemodynamic changes with enalapril in pulmonary arterial hypertension secondary to congenital heart disease. AB - Enalapril was used to treat five patients with pulmonary arterial hypertension secondary to congenital cardiopathy, three with ventricular septal defect, one with arterial septal defect, and one with patent ductus arteriosus. The dose of enalapril was 20 mg/day. All patients underwent pretreatment and posttreatment cardiac catheterization. It was concluded that enalapril may be a useful drug in the treatment of pulmonary arterial hypertension secondary to congenital cardiopathy. PMID- 3028718 TI - T-lymphocytes subpopulation before and after surgical removal of hepatocellular carcinoma. AB - This study included 16 histologically confirmed hepato-cellular carcinoma (HCC) patients. At the same time, 32 normal healthy individuals and 20 asymptomatic hepatitis B surface antigen (HBsAg) carriers were served as control groups. The study disclosed that T cells, OKT4 cells and the ratio of OKT4 cells/OKT8 cells in peripheral venous blood of HCC patients before surgery were lower than that of either normal control or asymptomatic HBsAg carriers. The difference was statistically significant. The T cells, OKT4 cells and the ratio of OKT4 cells/OKT8 cells of 4 HCC patients with positive HBeAg did not show further decrease as compared with that of 12 HCC patients with negative HBeAg. Periodic examination of T cells, OKT4 cells and OKT8 cells in 16 HCC patients after surgical removal of hepatocellular carcinoma tumor revealed that preoperative depressed T cells will decline further in the first postoperative week, but it will gradually increase to near normal range in the fourth or fifth postoperative week. However, OKT4 cells still remained in depressed state during the postoperative observation period. In the conclusion, it has been suggested that the derangement of T-cell subsets observed in HCC patients is not related to either HBsAg or even HBeAg. It can be either precondition to HCC formation or further follow up period being needed to look for the change of OKT4 cells. PMID- 3028720 TI - [Natural killer cell activity in patients with nasopharyngeal carcinoma]. AB - In 1984 and 1985, a total of 30 patients with biopsy confirmed nasopharyngeal carcinoma (NPC) were randomly selected for studies on natural killer cell (NK) activity. Control study on 30 normal subjects were also done. Lymphocyte separation was done by centrifuge of heparinized peripheral blood with Ficoll Hypaque and monocytes were removed by Petri dish adhesion. K562 cells were used as target cells of NK activity. The lymphocyte-target reaction of 4 hours was done and the ratio was 50:1, 25:1 and 12.5:1. The results of 50:1 was taken as the NK activity. The mean value of NK activity in NPC patients was 34.8 +/- 22.7%, this is significantly different (t = 2.90, p less than 0.01) from 51.6 +/- 22.1% of the controls. If NK activity of 20% or more are taken for the normal standard, then the positive rate in NPC patients was 63.3% and that in the controls was 90.0%. The difference is significant, as chi 2 = 4.56, p less than 0.05. The NK activity in NPC patients is not correlated with the sex and age of the patients, the disease extent and the EB virus associated antibody titers. PMID- 3028719 TI - Circulating immune complexes in the sera and ascites of hepatocellular carcinoma or chronic hepatitis patients. AB - Circulating immune complexes (CIC) in the sera or ascites of hepatocellular carcinoma (HCC), chronic hepatitis patients and normal healthy persons were measured by polyethylene glycol (PEG) and C1q solid-phase microassay (C1q-SPMA). Both the PEG and C1q-SPMA methods showed the serum CIC levels of HCC patients were significantly higher than those of chronic hepatitis patients and of normal persons. The CIC levels of chronic hepatitis patients were also significantly higher than those of normal persons as detected by PEG method but not by C1q SPMA. The ascites from HCC patients also had CIC. But the amount of CIC in ascites was significantly lower than those of the serum from the same HCC patients. These results suggest that the increase of CIC may play some pathological role in the HCC patients. PMID- 3028721 TI - How antidepressants work: cautionary conclusions based on clinical and laboratory studies of the longer-term consequences of antidepressant drug treatment. AB - Time-dependent alterations in the functional activity of adrenergic and serotonergic neurotransmitter systems and, in particular, a frequently observed down-regulation of brain beta-adrenoceptors have been implicated in antidepressant drug effects. Current studies of catecholamine and serotonin neurotransmitter systems suggest that the net physiological output changes in neuroendocrine responses, blood pressure, sleep and motor activity which follow various antidepressant treatments in psychiatric patients, normal controls and different experimental animals are not indicative of a common response pattern to all therapeutically effective agents. Rather, antidepressant treatment effects differ according to many variables, including the pre-existing state of the organism (e.g. depressed, stressed or normal), the species, the duration of treatment and the particular brain or peripheral circuits investigated. Examples are cited from our studies of the effects of monoamine oxidase inhibitors and other antidepressants on noradrenergic-serotonergic interactions that affect melatonin release and other neuroendocrine responses, on some additional functional end-points, and on depressive mood and other symptoms in patients with depression or other tricyclic-responsive disorders. These examples illustrate the complexity found in attempts to identify a unitary mechanism of antidepressant drug action. PMID- 3028722 TI - Neuroendocrine and other studies of the mechanism of antidepressant action of desipramine. AB - It is not known whether in depressed patients antidepressant treatment increases or reduces monoaminergic neurotransmission. Clinical studies are therefore reviewed that investigate adaptive changes at adrenoceptors in depressed patients treated with desipramine, and the net effect of these changes upon neurotransmission. Although in animals chronic desipramine treatment enhances the responsiveness of alpha 1-adrenoceptors to phenylephrine, no such effect could be demonstrated in patients upon the responsiveness of pupil diameter to phenylephrine. However, in keeping with animal studies, clinical evidence of altered responsiveness of alpha 2-adrenoceptors could be demonstrated after chronic desipramine treatment. The alpha 2-mediated growth hormone response to clonidine was increased after one week's treatment with desipramine and then reduced during the second and third weeks of treatment. No clinical measure of the responsiveness of central beta-adrenoceptors is available. However, the secretion of melatonin is a measure of neurotransmission at noradrenergic terminals in the pineal with alpha 1-, alpha 2- and beta 1-adrenoceptors. In normal volunteers the secretion of melatonin was increased by the noradrenaline uptake inhibitors desipramine and (+)-oxaprotiline; (-)-oxaprotiline had no effect. In depressed patients melatonin secretion was increased after three weeks' treatment with desipramine. These and other clinical studies suggest that antidepressant treatments increase noradrenergic neurotransmission in depressed patients. PMID- 3028723 TI - Effect of repeated administration of clenbuterol on the regulation of beta adrenoceptors in the central nervous system of the rat. AB - The effect of the centrally acting beta-adrenoceptor agonist clenbuterol on beta adrenergic responsiveness, beta-receptor density and N-protein coupling was studied in the rat cerebral cortex (which contains primarily beta 1-receptors) and cerebellum (containing mostly beta 2-receptors). The objective was to determine whether clenbuterol's effect on these variables was similar to that produced by standard antidepressants. When given to rats repeatedly, clenbuterol caused a decrease in beta-adrenergic responsiveness in slices from either the cerebral cortex or the cerebellum. The decreased beta-responsiveness in the cerebellum was associated with a decrease both in the density of beta-receptors and in receptor/N-protein coupling. In the cortex, only reduced receptor/N protein coupling was observed by in vitro ligand-binding methods. However, when quantitative autoradiography was employed, clenbuterol treatment was found to reduce the binding of [125I]iodopindolol to beta 2-receptors throughout the brain, whereas binding to beta 1-receptors was not reduced. The down-regulation of beta 2-receptors by clenbuterol is due to its acting centrally as a beta 2 agonist. Although clenbuterol has about an equal affinity for beta 1-receptors and beta 2-receptors, no evidence was found for agonist activity of this drug at beta 1-receptors in the cerebral cortex. The strategies described here should be helpful in investigating important properties of centrally acting beta-agonists that might have potential as antidepressants. PMID- 3028724 TI - Depression in an animal model: focus on the locus ceruleus. AB - When rats are exposed to highly stressful events over which they have no control, they subsequently show many of the symptoms seen in depression in humans. In the attempt to discover neurochemical factors underlying depression, the neurochemical basis of stress-induced behavioural depression in rats has been studied extensively. Initial research (1968-1976) indicated that behavioural depression in this model was produced by alteration of noradrenaline (NA) concentrations in the brain. More recent research has indicated that the critical change may be a large depletion of NA in the locus ceruleus (LC). Behavioural depression may result when such NA depletion is sufficient to reduce NA release in the LC region, leading to a 'functional blockade' of inhibitory alpha 2 receptors in that brain region. Studies have now shown that behavioural depression after uncontrollable shock can be mimicked by pharmacological blockade of alpha 2-receptors in the LC region. Conversely, behavioural depression can be eliminated by either infusion of clonidine into the LC to replace at the alpha 2 receptors the NA depleted after uncontrollable shock, or infusion of pargyline into the LC to prevent the depletion of NA that otherwise follows uncontrollable shock. If alpha 2-receptors are functionally blocked in depression, then release of NA in regions innervated by the LC should be increased and stimulation of postsynaptic adrenoceptors outside the LC should be higher than normal. Thus, higher-than-normal stimulation of postsynaptic NA receptors should also produce behavioural depression; this has been demonstrated. PMID- 3028725 TI - Effects of chronically administered antidepressants and electroconvulsive treatment on cerebral neurotransmitter receptors in rodents with 'model depression'. AB - The aim of this study was to develop a model of depression in laboratory animals in which chronically administered antidepressant drugs and electroconvulsive treatment (ECT) would produce receptor effects similar to but more marked than those in normal animals. The models discussed in detail are reserpinized and centrally chemosympathectomized rats. Other models currently under investigation are albino Swiss mice that respond with motor inhibition to high doses of morphine and rats tolerant to morphine. The reserpine model seems to be of some value, because in reserpinized rats antidepressants and ECT lead to adrenoceptor changes the same as or more marked than those observed in normal animals. Central chemosympathectomy with 6-hydroxydopamine prevents several receptor actions of imipramine, though not of ECT. The 'opiate models', though apparently not very promising, need further study. PMID- 3028726 TI - The effects of electroconvulsive therapy and antidepressant drugs on monoamine receptors in rodent brain--similarities and differences. AB - Repeated administration to rodents of electroconvulsive shock (ECS) produces changes in brain monoamine biochemistry and function, several of which are also seen after repeated administration of antidepressant drugs. Both repeated ECS and antidepressant drug administration decrease cortical beta-adrenoceptor density and attenuate the alpha 2-adrenoceptor-mediated sedation response to clonidine injection. Neither procedure alters phenylephrine-induced locomotor activity in mice, a measure of alpha 1-adrenoceptor function. Most antidepressant drugs decrease type 2 5-hydroxytryptamine (5-HT2) receptor density in frontal cortex and 5-HT2 receptor-mediated head-twitch behaviour in mice. In contrast, repeated ECS increases both 5-HT2 receptor density and the head-twitch response, making it difficult to propose any simple hypothesis linking changes in this receptor with antidepressant activity. The putative agonist for the 5-HT1A receptor 8-hydroxy-2 (di-n-propylamino) tetralin (8-OH-DPAT) produces a hypothermic response in mice, apparently by acting as an agonist at presynaptic 5-HT1 receptors. Repeated administration of antidepressant drugs and lithium markedly attenuates this hypothermic response. Repeated ECS also attenuates this response, the attenuation lasting for at least 20 days after the last ECS. Repeated ECS, but not antidepressant drug administration, markedly enhances dopamine-mediated behaviour. While the similarities in action between ECS and antidepressant drugs may help explain the therapeutic action of electroconvulsive treatment, the differences may provide clues to the efficacy of this treatment in drug-resistant depressive illness. PMID- 3028727 TI - Beta-adrenoceptor function in human adult skin fibroblasts: a study of manic depressive illness. AB - Catecholamine theories of affective illness provide a rationale for the study of beta-adrenoceptor function in these disorders. Skin fibroblasts were grown in tissue culture from skin biopsies of normal volunteers and manic-depressive subjects for measurement of isoprenaline-stimulated production of cyclic AMP. Monolayers of fibroblasts in wells were incubated for 3 min with or without 0.5 microM-isoprenaline. The cyclic AMP was isolated by ion-exchange chromatography and quantitated by radioimmunoassay. The isoprenaline-stimulated levels of cyclic AMP in manic-depressive subjects (n = 12) were no different from those in normal volunteers (n = 13). Thus, no evidence was found for abnormal beta-adrenoceptor function in manic-depressive illness. PMID- 3028728 TI - Genetic studies at the receptor level: investigations in human twins and experimental animals. AB - In receptors, as in enzymes, quantitative as well as qualitative genetic variation may exist. Studies in inbred strains of mice have shown for various receptors that the receptor density as determined by Bmax values is under genetic control. In healthy adult twins we have shown that the density of alpha adrenoceptors on platelets is also influenced by genetic factors, since monozygotic twins were much more similar to one another than dizygotic twins. However, Bmax values are up-regulated and down-regulated by endogenous neurotransmitters and pharmacologically active agents. Thus, receptor densities are under considerable regulatory influences. Bmax values therefore reflect regulatory mechanisms rather than innate characteristics of the receptor protein. In another twin study we failed to find evidence for a genetic influence on the density of imipramine-binding sites on platelets. Since qualitative variation (polymorphism) is well known in enzymes, it may also apply to receptors. Qualitative differences in the receptor protein within one species would be of particular interest because of possible functional implications. As a first approach we examined central benzodiazepine receptors by photoaffinity labelling and sodium dodecyl sulphate-polyacrylamide gel electrophoresis. A comparison of fish, frog, chicken, mouse, rat and calf led to the detection of variation between species. Investigations in five inbred mouse and rat strains have not so far revealed genetic variation in benzodiazepine receptors. Nevertheless variation may be detectable by more sensitive methods such as peptide mapping after limited proteolysis or two-dimensional electrophoresis. PMID- 3028729 TI - Platelet radioligand binding and neuroendocrine challenge tests in depression. AB - The purpose of this work was to examine the number and function of alpha 2 adrenoceptors and the number of serotonin uptake sites in depressed patients and controls. Platelet alpha 2-adrenoceptors and platelet serotonin uptake sites were labelled with [3H]yohimbine and [3H]imipramine respectively. Central alpha 2 adrenoceptor function was assessed by growth hormone and other responses to challenge with the alpha 2-agonist clonidine. No overall difference in the binding parameters was observed between the control and depressed groups, but the results highlight the importance of drug-free interval, menopausal status and membrane protein concentration within the binding assays in the interpretation of such studies. The growth hormone response to clonidine tended to be blunted in depressed females and was significantly blunted in the subgroup of depressives who failed to suppress plasma cortisol concentrations in response to dexamethasone. Depressed subjects also showed a smaller decrease in diastolic blood pressure and a smaller increase in sedation than control subjects. PMID- 3028730 TI - [Early diagnosis and reoperation for local recurrence after radical operation for rectal cancer]. PMID- 3028732 TI - [Giant cell tumor of the tendon sheath: report of 36 cases]. PMID- 3028731 TI - [Intestinal polyposis]. PMID- 3028733 TI - [Surgical treatment of early gastric carcinoma]. PMID- 3028734 TI - Acyclovir in the treatment of infection due to herpes viruses. AB - Intravenous infusions of acyclovir were administered to 15 patients suffering from herpes simplex or varicella-zoster infections. The therapeutic effect of the drug consisted in rapid cessation of progression of the disease and in considerable shortening of duration of clinical symptoms. There were no adverse reaction in our groups of patients. There were no relapses after the treatment with acyclovir had been stopped. From what has been stated above, we take acyclovir for a highly efficient and safe drug for therapy of HSV and VZV infections. PMID- 3028735 TI - Acute pancreatitis as presenting symptom and sole manifestation of small cell lung carcinoma. AB - A 58-year-old man with no sign of pulmonary disease and a normal chest x-ray presented with acute pancreatitis resistant to conventional medical management and a mass in the head of the pancreas. The presumptive diagnosis was pancreatic cancer with tumor-induced pancreatitis. However, endoscopic retrograde cholangiopancreatography suggested metastatic rather than primary tumor, so that an extrapancreatic primary was actively sought. Further lung work-up demonstrated a small cell carcinoma of the lung. This case indicates that metastasis-induced acute pancreatitis can be the presenting symptom and sole manifestation of lung cancer. PMID- 3028737 TI - Studies of the structure and functional organization of foreign DNA integrated into the genome of Nicotiana tabacum. AB - In transgenic plants obtained either by Agrobacterium tumefaciens-mediated transformation or by direct DNA transfer, the structure of integrated chimeric donor DNA remains stable during vegetative proliferation, during sexual transmission, and under various selection conditions. We correlate the level of expression of the introduced gene in independent transformants and their offspring with the particular arrangement and modification of their integrated DNAs. Genetic analysis of transgenic plants shows that the chimeric gene is transmitted in a Mendelian fashion to the F1 and F2 progeny as a single dominant trait. Deviations from the expected segregation pattern are discussed with respect to different levels of gene activity. We compare the gene activity in heterozygotes versus homozygotes, and show variation in activity between plants regenerated independently from the same transformed callus. Cotransformation studies with two physically unlinked and partly homologous plasmids carrying two different marker genes indicate that they are physically linked after integration into the host genome. PMID- 3028736 TI - Differential transient and long-term expression of DNA sequences introduced into T-lymphocyte lines. AB - We have used a protoplast fusion protocol to introduce the genes encoding neomycin phosphotransferase (neo) and chloramphenicol acetyltransferase (CAT) into murine and human T-lymphocyte lines. Plasmid constructs containing the neo gene under the control of the promoters from the Rous sarcoma virus long terminal repeat (RSV LTR), the SV40 early region, or the herpes simplex virus thymidine kinase gene (HSV TK) can stably transform each of three T-cell lines to G-418 resistance. The characteristic frequencies for different cell lines can differ by at least two orders of magnitude, although initial DNA uptake and transient expression are similar. In the two murine cell lines, low numbers of gene copies are retained in long-term transformants. Prior to integration, transient expression assays for cat or neo gene products reveal that the differences in intrinsic promoter strength of different constructs are further influenced by the coding sequences being transcribed. Thus, while transient expression of the neo protein is similar from both the Rous LTR and the SV40 early promoter, the Rous LTR directs synthesis of CAT protein at levels two orders of magnitude higher than those from the SV40 early promoter. PMID- 3028738 TI - Improved secretion of heterologous proteins by Saccharomyces cerevisiae: effects of promoter substitution in alpha-factor fusions. AB - The effects of promoter strength on secretion of a heterologous protein, somatomedin-C (SMC), by the yeast Saccharomyces cerevisiae were studied by using the promoters of the MF alpha 1, ACT, and CYC1 genes to control expression of alpha-factor/SMC gene fusions. When a low-copy centromere vector was used to carry the gene fusions in yeast transformants, the greatest secretion was obtained with the MF alpha 1 promoter construction and the least with the CYC1 promoter construction. Unexpectedly, using two types of multicopy vectors, the greatest secretion was obtained with the CYC1 promoter construction and the least with the MF alpha 1 promoter construction. The decrease in secretion by the strongest promoter construction (MF alpha 1 promoter) on multicopy vectors was associated with a decrease in SMC mRNA during growth, a decrease in vector copy number, a decrease in vector stability, and a decrease in transformation frequency. The results demonstrate that, unlike in intracellular expression, promoter strength is not simply related to secretion expression levels. Selection against oversecreting cells during growth may explain the reduced secretion efficiency of strong promoter constructions. PMID- 3028739 TI - Pancreatic phospholipase A2: isolation of the human gene and cDNAs from porcine pancreas and human lung. AB - Oligonucleotide probes synthesized using the published protein sequence for porcine pancreatic phospholipase A2 (PLA2; Puijk et al., 1977), were used to screen a cDNA library made from porcine pancreas. One resulting clone which encoded the entire porcine PLA2 enzyme was then used to screen various human cDNA and gene libraries. A 562-bp cDNA clone from human lung and two overlapping genomic clones encoding the identical transcript were obtained. These clones encoded a 126-residue peptide corresponding to human "pancreatic" PLA2. The gene spanned 4.9 kb and contained three introns of 1692, 800, and about 2650 bases, respectively. Hybridization of the human cDNA against blots of total human DNA detected the same set of fragments contained in both genomic clones and failed to detect more than one distinct gene. Under equivalent conditions, the same probe detected two discrete bands in cobra (Naja naja) genomic DNA. Apparently, there is a single or small number of identical human genes encoding "pancreatic" PLA2 which are transcribed also in lung tissue; human pancreatic PLA2 appeared more closely related at the nucleotide level to snake counterparts than to other reported human PLA2 enzyme forms. PMID- 3028740 TI - Vectors for expression and amplification of cDNA in mammalian cells: expression of rat phenylalanine hydroxylase. AB - We have constructed two recombinant plasmid vectors for direct expression and amplification of cDNA in mammalian cells. Each vector carries two dominant selectable markers (the bacterial neo gene and the mouse DHFR gene), a promoter sequence (viral LTR in pAV009/A+, and sheep metallothionein promoter in pMT010/A+), a polyadenylation signal sequence, and a Bam HI site to allow insertion of cDNA. We have used these vectors to prepare recombinant clones for the expression of rat phenylalanine hydroxylase (PH) in LTK- cells. Selection of transformants with neomycin followed by selection of the transformants in methotrexate led to a 30- to 60-fold amplification of the DHFR marker and co amplification of the PH cDNA, with a corresponding increase in the level of PH mRNA and enzyme polypeptide. The expressed enzyme has a subunit molecular weight of 50,000 which corresponds to the W- allele of rat liver PH. PH activity was detected in the transfected cells by enzymatic measurement of the conversion of [14C]phenylalanine to [14C]tyrosine, and by growth of these cells in a tyrosine free culture medium. Expression of rat PH in cell culture should facilitate the analysis of the biochemical properties of this enzyme. PMID- 3028741 TI - Re: Concept of virus as an etiological agent in the development of insulin dependent diabetes mellitus. PMID- 3028742 TI - The effect of non-esterified fatty acids on vascular ADP-degrading enzyme activity. AB - Following our observations that non-esterified fatty acids (NEFAs) inhibit prostacyclin (PGI2) synthesis and accelerate PGI2 degradation, we have examined the possibility that NEFAs may also affect the activity of vascular ADPase, which converts the platelet pro-aggregatory adenosine diphosphate (ADP) to adenosine, an inhibitor of aggregation and a vasodilator. Incubation, in buffer solutions, of NEFAs with intact rat aortic rings significantly inhibited vascular ADPase activity. This inhibition was more marked at higher NEFA concentrations and with unsaturated fatty acids (linoleic, oleic) than with a saturated fatty acid (stearic). This NEFA-mediated inhibition of vascular ADPase activity could be prevented by the prior addition of fatty acid-free human albumin to the incubate. Similarly, the vascular rings recovered from NEFA-mediated inhibition by washing and further incubation in NEFA-free buffer. Therefore, elevated NEFA concentrations inhibit, reversibly, an enzyme system which is thought to protect the vascular endothelium. The NEFA-mediated inhibition of ADPase activity was also confirmed following incubation of rat aortic rings in human serum enriched with exogenous NEFA. These findings provide further evidence that NEFAs may contribute to the pathogenesis of vascular disease associated with diabetes mellitus and of other conditions where an elevation of serum NEFA concentrations occurs. PMID- 3028743 TI - [Virus-like particles containing the MDG sequences in cultured media of Drosophila cell lines]. PMID- 3028745 TI - [Sensitivity to hydrogen peroxide and in vivo survival of Syrian hamster cells transformed in vitro with Rous sarcoma virus]. PMID- 3028744 TI - [Molecular organization of the complete gene encoding a human cellular tumor antigen (p53)]. PMID- 3028746 TI - Audiogenic seizures during ethanol withdrawal can be blocked by a delta opioid agonist. AB - In rats the influence of the delta opioid agonists [Leu]enkephalin, [D-Ala2-D Leu5]enkephalin, [D-Ala2]methionine enkephalinamide and synthetic analogue of [Met]enkephalin: Tyr-D-Ala-Gly-Phe-D-Leu-OMe on audiogenic seizures was tested during ethanol abstinence. All investigated drugs significantly inhibited this ethanol withdrawal symptom. The results are compatible with the hypothesis of opioid involvement in the ethanol abstinence syndrome. PMID- 3028747 TI - Effects of long-term ethanol consumption on GABAergic neurons in the mouse hippocampus: a quantitative immunocytochemical study. AB - The effects of 6 months' ethanol consumption by mice on hippocampal GABAergic neurons were investigated by means of an immunocytochemical method using GABA antibodies. Although ethanol treatment did not modify body or brain weights in our experimental conditions, two differences were observed in ethanol-treated mice, as compared to controls: a decrease in the labelling intensity of immunopositive neurons and fibers in the dorsal and the ventral parts of the hippocampus; and a decrease in the number of immunopositive neurons. This neuronal loss was statistically significant in the ventral hippocampus only, where it reached about 25% in the stratum radiatum. It is concluded that chronic ethanol consumption leads to a decrease in GABA content of hippocampal neurons and to a loss of GABAergic neurons, mostly in the ventral part of the hippocampus. These alterations in GABAergic transmission could be related to the well known functional deficits observed in chronic alcoholism. PMID- 3028748 TI - [Hereditary neuropathy with liability to pressure palsies. A contribution to the differential diagnosis of multiplex mononeuropathy]. AB - Hereditary neuropathy with liability to pressure palsies could be diagnosed in two families. This little-known, dominantly inherited disorder is clinically characterised by recurring, spontaneously regressive palsies of peripheral nerves, mostly after minimal mechanical compression of the nerve concerned. It can be clearly differentiated from mononeuropathies of different pathogenesis even in clinically not involved nerves, by means of pathological neurographic findings and by the detection of pathognomonic myelinic thickenings in nerve biopsies. Prognosis is favourable if recurrences are avoided. This means that counselling of the patients and their families is of prophylactic significance. PMID- 3028749 TI - [Prophylaxis of hepatitis A]. PMID- 3028750 TI - [Aspergillosis following liver transplantation as a hospital infection]. AB - In the first four weeks after a liver transplantation, there was an invasive aspergillosis with a lethal course in three out of five patients who were treated postoperatively in the same room. The clinical symptoms were very different. One patient was asymptomatic, and the diagnosis could only be made by autopsy. In another patient, pulmonary symptoms, and in the third patient, cerebral symptoms were the most prominent. In the two latter patients, the infection was demonstrated in the sputum and by bronchoalveolar lavage. The disease course was fulminant in all patients, and therapy was without success. Owing to this high incidence, mycological investigations were carried out on the ward. A flower bench in the hall beside the ward was probably the main focus of distribution. To avoid such nosocomial infections, foci of aspergillus distribution should if possible be removed from the surroundings of patients with weakened immune resistance. PMID- 3028751 TI - [After care in differentiated thyroid carcinoma]. PMID- 3028752 TI - Granular-cell myoblastoma of the cricopharyngeal muscle. PMID- 3028753 TI - [Filling periodontal bone pockets with porous hydroxylapatite (Osprovit)]. PMID- 3028754 TI - [In vitro research on the action of different types of anthophyllite]. PMID- 3028756 TI - [Amplification of c-myc proto-oncogene in primary tumors, metastases and blood leukocytes of patients with ovarian cancer]. AB - The structure of DNA sequences homologous to the viral myc oncogene from ovarian tumours, intact tissues and the same patients peripheral blood leukocytes has been studied. The Southern blot analysis of malignant tumour DNA reveals amplification of c-myc protooncogene of 3 from 11 patients. Metastases DNAs of two from those patients also contain multiple copies of c-myc. No myc gene amplification has been detected in 6 examined benign ovarian tumours. The presence of extra copies of myc-related sequences has been shown in two from three DNA samples from peripheral blood leukocytes of patients with ovarian cancer. PMID- 3028755 TI - [Characteristics of genome sequences in mouse spleen cells activated in virus induced Rauscher leukemia]. AB - The restriction analysis of BALB/c and AKR mice genome DNA was used to show the translocation and amplification of gene which is expressed with the Rauscher erythroleukemia as the 35S nuclear RNA. The homology of this RNA to 10 defined oncogenes was studied. It was found that the cDNA 35S RNA is efficiently hybridized only with v-sis oncogene. The clones homologous to the 35S RNA, v-sis oncogene and 3'-area of the Rauscher leukemia virus genome are isolated from the genes' library of mice erythroleukemic cells. PMID- 3028758 TI - [Effect of vaccination of mice with endogenous retrovirus on the development of tumors induced by gamma-irradiation or 7,12-dimethylbenz[a]anthracene]. AB - Mouse vaccination with alive endogenous N-tropic virus OA-3 inhibited and decreased the development of the Rauscher leukemia in C57B1/6 mice (B-type) and SWR mice (N-type) as well as the development 7,12-dimethyl benzanthracene (DMBA) induced tumours in mouse hybrids (neither N-, nor B-types). The effect of vaccination was DMBA- or MLV-P-dose-dependent. Vaccination with the same virus did not affect the incidence of gamma-irradiation-induced leukemia in CBA mice (N type) and C57B1/6 mice while it increased twice the incidence of radiation leukemia in DBA mice (N-type). However, the incidence of thymomas lowered in radiation leukemia-bearing vaccinated mice of all the 3 strains, which may result from inhibition of murine thymotropic endogenous virus reproduction. The data obtained indicate the participation of murine own endogenous viruses in DMBA- or gamma-irradiation induced carcinogenesis. PMID- 3028757 TI - [Cytotoxic activity and activation ability of Syrian hamster NK cells isolated in the Percoll density gradient]. AB - Natural killer cells possessing cytotoxic activity (CTA) and the ability to be activated by an interferon inductor and Newcastle disease virus, were isolated from blood, spleen and bone marrow of Syrian hamsters in the discontinuous Percoll density gradient. According to morphological criteria these cells were identified as large granular lymphocytes (LGL). The largest amount of LGL was isolated in the fraction containing 52-55% Percoll. CTA of cells from this fraction was the highest as compared to nonfractionated control or cells from other Percoll fractions. Effective (3-fold) activation of bone marrow cells isolated in the fraction of 49% Percoll with initially low CTA suggests that this Percoll fraction contains noncytotoxic precursors of NK cells. PMID- 3028759 TI - Electrophysiological pattern of development of muscle fatigue in patients undergoing dialysis. PMID- 3028760 TI - Multiple role of peripheral neuropathy in the pathogenesis of the so-called "diabetic foot". Clinical and electromyographical (EMG) study of 42 cases of "mutilating acropathy". PMID- 3028761 TI - Diagnostic value of automatic electromyographic analysis in the assessment of degree of damage in neuropathy. PMID- 3028762 TI - Examination of the peripheral sensory neurone in subjects with alcohol dependence syndrome. PMID- 3028763 TI - A late motor response: the peripheral late wave (PWL). PMID- 3028764 TI - [The CRF test in psychiatry. Preliminary studies]. AB - The HPA axis exploration in the endogenous depressions indicates an hyperactivity of this system. But the reason of this perturbation is not definite. The corticotropin-releasing factor (CRF) test has been described from research work carried out by Vale et al. In the endogenous depressions, this test enables us to specify physiopathological factors of the HPA axis dysfunction. Some research suggest an hypothalamic and limbic CRF hypersecretion in the endogenous depressions. But the results are not consistent principally because of the diversity of technical protocols. So the authors try to take stock of the CRF test specificity. They report the results obtained after 100 micrograms o-CRF intravenous administration in 7 voluntary normal men. PMID- 3028765 TI - Dynamic nuclear polarisation of biological matter. AB - Polarised targets as used in high energy physics experiments may be of considerable interest in biological structure research using polarized neutrons. So far, this promising method has been facing difficulties in getting reasonable polarization of the target nuclei. We report on a polarized "frozen spin" target which has been prepared from an enzyme dissolved in a mixture of heavy water and deuterated propanediol doped with a completely deuterated paramagnetic radical. Clusters of 700 protons defined by the structure of lysozyme embedded in a fully deuterated matrix were polarized to 75% within an hour by 4 mm microwave irradiation in a magnetic field of 2.5 tesla at a temperature of 0.3 K. The polarisation behaviour of biological targets can be compared to the best frozen spin target materials in high energy physics research. PMID- 3028766 TI - Hyperoxia-induced converting enzyme insufficiency in conscious rat: cardiovascular effects. AB - The effects of exposing rats to hyperoxia (100 percent O2) at normal atmospheric pressure for periods of 24-48 hours on components of the renin-angiotensin system and on blood pressure control were examined. Intrapulmonary conversion of angiotensin I (AI) to angiotensin (AII) was assessed using an isolated lung preparation perfused at constant flow. Exposure of rats to hyperoxia for 44-48 hours reduced single pass conversion of AI to AII in the pulmonary circulation from control levels of 82 +/- 4 to 29 +/- 5 percent (p less than 0.001). AII levels in trunk blood of 44-48 hour O2 exposed animals were 5.2 +/- 1.9 pg/ml, compared to 37.9 +/- 10.0 pg/ml in controls (p less than 0.001). Mean arterial pressure decreased significantly from 117 +/- 4.3 to 103 +/- 6.7 mmHg (p less than 0.05) in the O2 exposed group despite a threefold increase in plasma renin activity. The pressor response to exogenous AI was significantly diminished by O2 exposure, while the pressor response to exogenous AII remained unchanged from control. Pulmonary angiotensin-converting enzyme activity fell to approximately 50 percent of control in O2 exposed animals, but circulating converting enzyme activity was not change in this group. None of these alterations was apparent following 24 hours of hyperoxic exposure. These data suggest that O2 induced impairment in activity of angiotensin-converting enzyme at the endothelial membrane level has functionally significant effects on cardiovascular homeostasis, probably via reduced generation of endogenous AII. PMID- 3028767 TI - Stability and effectiveness of chlorine disinfectants in water distribution systems. AB - A test system for water distribution was used to evaluate the stability and effectiveness of three residual disinfectants--free chlorine, combined chlorine, and chlorine dioxide--when challenged with a sewage contaminant. The test distribution system consisted of the street main and internal plumbing for two barracks at Fort George G. Meade, MD. To the existing pipe network, 152 m (500 ft) of 13-mm (0.5 in.) copper pipe were added for sampling, and 60 m (200 ft) of 2.54-cm (1.0 in.) plastic pipe were added for circulation. The levels of residual disinfectants tested were 0.2 mg/L and 1.0 mg/L as available chlorine. In the absence of a disinfectant residual, microorganisms in the sewage contaminant were consistently recovered at high levels. The presence of any disinfectant residual reduced the microorganism level and frequency of occurrence at the consumer's tap. Free chlorine was the most effective residual disinfectant and may serve as a marker or flag in the distribution network. Free chlorine and chlorine dioxide were the least stable in the pipe network. The loss of disinfectant in the pipe network followed first-order kinetics. The half-life determined in static tests for free chlorine, chlorine dioxide, and combined chlorine was 140, 93, and 1680 min. PMID- 3028768 TI - Quantitation and identification of organic N-chloramines formed in stomach fluid on ingestion of aqueous hypochlorite. AB - The chemical reactions that hypochlorite undergoes in the body when chlorinated water is ingested have received very little attention. Because amino nitrogen compounds are important components of the average diet, the reactions of hypochlorite with amino compounds in the stomach were investigated. Stomach fluid was recovered from Sprague-Dawley rats that had been fasted for 48 hr and administered 4 mL deionized water. The chlorine demand of the stomach fluid was determined. An average volume-independent demand of 2.7 mg chlorine was measured. At doses below 40 mg/L chlorine reducing reactions appeared to account for reduction of all oxidizing species within 15 min as measured by the FAS-DPD titrimetric method. At least part of the chlorine demand is associated with amino acids present in the stomach fluid. Amino acids were identified and quantified in the stomach fluid by precolumn derivatization with o-phthalaldehyde and high pressure liquid chromatography (HPLC). When stomach fluid is chlorinated to concentrations of chlorine between 200 and 1000 mg/L, organic N-chloramines are formed. After derivatization of chlorinated stomach fluid with dansyl sulfinic acid, fluorescent derivatives of chloramines were separated by HPLC. Three chloramino acid derivatives, N-chloroalanine, N-chloroglycine, and N chlorophenylalanine, were identified by cochromatography with known standards using two chromatographic methods. The yield of a chloramine that would form in stomach fluid on administration of hypochlorite to animals was determined using tritiated piperidine and doses of 200 and 1000 mg/L chlorine. Yields of tritiated N-chloropiperidine in recovered stomach fluid were 70% and 42%, respectively, of the theoretical amount expected. PMID- 3028769 TI - Epidermal hyperplasia in mouse skin following treatment with alternative drinking water disinfectants. AB - Female SENCAR mice were treated with aqueous solutions of hypochlorous acid (HOCl), sodium hypochlorite (NaOCl), chlorine dioxide (ClO2), and monochloramine (NH2Cl) by whole body exposure (except head) for a 10-min period for 4 days in the first experiment and for 1 day (except NH2Cl) in the second experiment. Animals were sacrificed the day following the last treatment (experiment 1) or on day 1, 2, 3, 4, 5, 8, 10, and 12 following treatment (experiment 2), and skin thickness was measured by light microscopy at X400 by use of an eyepiece micrometer. Concentrations of disinfectants were 1, 10, 100, 300, and 1000 mg/L, for experiment 1 and 1000 mg/L for experiment 2. Thickness of the interfollicular epidermis (IFE) for control animals was 15.4 +/- 1.5 micron. After 4 days of treatment at 1000 mg/L, HOCl and ClO2 increased thickness to 39 +/- 7.0 and 40.2 +/- 11.8, and NaOCl increased thickness to 25.2 +/- 6.1 micron. Only HOCl and ClO2 were tested at 300 mg/L, yielding an IFE thickness of 30.0 +/- 13.1 and 16.8 +/- 0.8 micron, respectively. The response to HOCl was found to be dose-related; the minimally effective dose was 100 mg/L. In earlier, preliminary tests to determine optimum treatment schedule, the response to HOCl appeared to be maximal after 4 days of treatment and tended to decrease with further treatment. The time course study following a single treatment of 1000 mg/L HOCl, however, showed a progression of IFE thickening of from 18.3 +/- 1.4 at 1 day to 30.8 +/- 8.0 at 8 days, decreasing to 19.1 +/- 6.2 micron at 12 days.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3028770 TI - Hypochlorous acid-activated carbon: an oxidizing agent capable of producing hydroxylated polychlorinated biphenyls. AB - Granular activated carbon (GAC), in the presence of dilute aqueous hypochlorite solutions typical of those used in water treatment, was converted to a reagent capable of carrying out free-radical coupling reactions and other oxidations of dilute aqueous solutions of phenols. The products included biphenyls with chlorine and hydroxyl substitution (hydroxylated polychlorinated biphenyls). For example, 2,4-dichlorophenol, a common constituent of wastewaters and also natural waters treated with hypochlorite, was converted to 3,5,5'trichloro-2,4' dihydroxybiphenyl and several related compounds in significant amounts. It is possible that these products pose more of a health hazard than either the starting phenols or the unhydroxylated polychlorinated biphenyl derivatives. PMID- 3028771 TI - Reconstitution of vesicle fusions occurring in endocytosis with a cell-free system. AB - We have used defined subcellular fractions to reconstitute in a cell-free system vesicle fusions occurring in the endocytic pathway. The endosomal fractions were prepared by immuno-isolation using as antigen an epitope located on a foreign protein, the transmembrane glycoprotein G (G-protein) of vesicular stomatitis virus. The G-protein was first implanted in the cell plasma membrane and subsequently endocytosed for 15 to 30 min at 37 degrees C. The endosomal fractions were immuno-isolated on a solid support using as antigen the cytoplasmic domain of the G-protein in combination with a specific monoclonal antibody. For comparative studies the plasma membrane was immuno-isolated from cells in the absence of G internalization with a monoclonal antibody against the exoplasmic domain of the G-protein. The immuno-isolated endosomal vesicles contained 70% of horseradish peroxidase internalized in the endosome fluid phase, exhibited an acidic luminal pH as shown by acridine orange fluorescence and differed in their protein composition from the immuno-isolated plasma membrane fraction. The fusion of endocytic vesicles originating from different stages of the pathway was studied in a cell-free assay using both a bio-chemical and a morphological detection system. These well defined endosomal vesicles were immuno isolated with the G-protein on the solid support and provided the recipient compartment of the fusion (acceptor). They were mixed with a post-nuclear supernatant containing endosomes loaded with exogenous lactoperoxidase (donor) at 37 degrees C. Fusion delivered the donor peroxidase to the lumen of acceptor vesicles permitting fusion-specific iodination of the G-protein itself. The fusion of vesicles required ATP and was detected only with an endosomal fraction prepared after internalization of the G-protein for 15 min at 37 degrees C but not with a plasma membrane or with an endosomal fraction prepared after 30 min G protein internalization. PMID- 3028772 TI - Establishment of 'normal' nervous cell lines after transfer of polyoma virus and adenovirus early genes into murine brain cells. AB - Brain cells from murine embryos were transfected with the polyoma virus large T or the adenovirus 5 EIA gene and, simultaneously, with the phosphotransferase coding NeoR gene. The efficiently transfected cells were selected for their resistance to Geneticin (G418) and their ability to clone at low cell density. Subsequently, most of the selected cells could be sub-cloned and continuously grown for 6-18 months so far. Their doubling time varied between 18 and 72 h. From independent transfections, more than one hundred cell lines were established. They did not exhibit a transformed phenotype, but subsequent transfection with the polyoma middle T gene induced their oncogenic transformation. The maintenance and expression of the transferred genes were verified. Most of the analyzed cell lines retained glial properties. These results suggest that the lines obtained as well as a further extension of this in vitro system should be of interest for the study of nervous cell interactions, differentiation and functions. PMID- 3028773 TI - Determination and analysis of the primary structure of the nerve terminal specific phosphoprotein, synapsin I. AB - A rat brain cDNA clone containing an open reading frame encoding the neuron specific protein synapsin I has been sequenced. The sequence predicts a protein of 691 amino acids with a mol. wt of 73 kd. This is in excellent agreement with the size of rat brain synapsin Ib measured by SDS--polyacrylamide gel electrophoresis. Inspection of the predicted primary structure has revealed the probable sites for synapsin I phosphorylation by the cAMP-dependent and Ca2+/calmodulin-dependent protein kinases. All of the biochemically observed intermediates of synapsin I digestion by collagenase can be verified by inspection of the sequence, and the collagenase-resistant fragment has been defined as the amino-terminal 439 amino acids of the molecule. Predictions of sequence secondary structure and hydrophobicity suggest that a central domain of approximately 270 amino acids may exist as a folded, globular core. The carboxyl terminal domain of the protein (the region sensitive to collagenase digestion) contains sites for Ca2+/calmodulin-dependent protein kinase phosphorylation. These sites are flanked by three regions of repeating amino acid sequence that are proposed to be the synaptic vesicle-binding domain of synapsin I. This region also shares homology with the actin-binding proteins profilin and villin. The characteristics of the synapsin I sequence do not support extensive homology with the erythrocyte cytoskeletal protein 4.1. PMID- 3028775 TI - Cloning of the human cDNA for the U1 RNA-associated 70K protein. AB - Anti-RNP sera were used to isolate a cDNA clone for the largest polypeptide of the U1 snRNP, a protein of mol. wt 70 kd designated 70K, from a human liver cDNA library constructed in the expression vector pEX1. The cro-beta-galactosidase-70K fusion protein reacted with various anti-RNP patient sera, a rabbit anti-70K antiserum, as well as with a monoclonal antibody specific for this protein. The sequences of four 70K peptides were determined and they match parts of the deduced amino acid sequence of the 1.3 kb insert of p70.1 indicating that it is a genuine 70K cDNA. Screening of a new cDNA library constructed from polysomal mRNA of HeLa cells with the p70.1 clone yielded an overlapping clone, FL70K, which was 2.7 kb long and covered the complete coding and 3'-untranslated sequence of the 70K protein in addition to 680 nucleotides upstream of the putative initiation codon, The predicted mol. wt of the encoded protein is approximately 70 kd. Amino acid analysis of the purified HeLa 70K protein yielded values close or identical to those deduced from the nucleotide sequence of the full-length cDNA. The 70K protein is rich in arginine (20%) and acidic amino acids (18%). Extremely hydrophilic regions containing mixed-charge amino acid clusters have been identified at the carboxyl-terminal half of the protein, which may function in RNA binding. A sequence comparison with two recently cloned RNA binding proteins revealed homology with one region in the U1 RNP 70K protein. This domain may also be responsible for RNA binding. PMID- 3028776 TI - Isolation of a murine Ly-6 cDNA reveals a new multigene family. AB - The Ly-6 alloantigens have been shown to play a critical role in T lymphocyte activation. To isolate a Ly-6 cDNA, synthetic oligonucleotides, based on the partial amino acid sequence of purified Ly-6E.1 protein, were used to probe a cDNA library. The synthetic oligonucleotides or the isolated cDNA detected a 1.1 kb RNA species. Sequence analysis of the cDNA clone revealed that the Ly-6E.1 protein consists of a 26-amino acid leader followed by a 108-residue, cysteine rich, core protein with no N-linked glycosylation sites. Southern blot analysis of genomic DNAs revealed multiple bands indicating a family of related genes. Using recombinant inbred and Ly-6 congenic strains of mice, restriction fragment length polymorphisms were demonstrable, and correlated with the Ly-6 allotype of the DNA donors. This probe will enable further molecular genetic analysis of the role of Ly-6-linked proteins in the process of T lymphocyte activation. Isolation of Ly-6 genomic clones may promote a further understanding of the complex tissue specific expression patterns characteristic of Ly-6-linked genes. PMID- 3028774 TI - A 22-kd protein (sorcin/V19) encoded by an amplified gene in multidrug-resistant cells, is homologous to the calcium-binding light chain of calpain. AB - We have previously shown that at least five linked genes are co-amplified and overexpressed in the multi-drug resistant (MDR) Chinese hamster ovary cell line CHRC5. We show here that one of these genes (class 4) codes for a small phosphorylated, cytosolic protein, sorcin/V19, known to be overproduced by many MDR cell lines. The class 4 gene codes for a nested set of mRNAs, varying in size between 1000 and 2500 nucleotides. Sequence analysis of complementary DNAs shows that these mRNAs encode a protein of 198 amino acids. The identity of this protein with sorcin was established by comparison with the amino acid sequence of two peptides from mouse sorcin. Hamster sorcin is a 22-kd protein with four 'E-F hand' structures typical of calcium-binding sites and it has substantial homology with the light chain of calpain. Two of the calcium-binding sites contain putative recognition sites for cAMP-dependent protein kinase. These may account for the known phosphorylation of sorcin. The unknown function of sorcin might therefore be controlled by both calcium and cAMP levels. The contribution of sorcin to multidrug resistance, if any, remains to be tested. PMID- 3028777 TI - Both Epstein-Barr virus (EBV)-encoded trans-acting factors, EB1 and EB2, are required to activate transcription from an EBV early promoter. AB - We have identified two Epstein--Barr virus (EBV) transacting factors which are involved in the transcriptional activation of EBV early promoters in latently infected Raji cells. In Raji cells, expression of the factor EB1 encoded by the open reading frame (ORF) BZLF1 is necessary and sufficient to disrupt latency. However, factor EB2 encoded by the ORF BMLF1- BSLF2 does not disrupt latency when expressed alone in Raji cells. Expression of an EBV activatable early promoter depends on the presence of both EB1 and EB2. PMID- 3028778 TI - Secondary genomic rearrangement events in pre-B cells: VHDJH replacement by a LINE-1 sequence and directed class switching. AB - We describe rearrangement events which alter expression from a productive VHDJH rearrangement in an Abelson murine leukemia virus-transformed pre-B cell line. One such rearrangement results in replacement of the initially expressed variable region gene by a site-specific join between the open reading frame of a LINE-1 repetitive element and a remaining JH segment. We discuss this event in the context of the 'accessibility' model of recombinase control, and with respect to similar rearrangements involved in oncogene activation. In another subclone of the same pre-B cell line, altered heavy chain expression resulted from a mu to gamma 2b class switch recombination which occurred by a recombination-deletion mechanism but involved a complex inversion. We provide evidence that the germline gamma 2b region is specifically expressed in pre-B cell lines and early in normal development. We propose that the predisposition of pre-B cell lines to switch to gamma 2b production may reflect a normal physiological phenomenon in which the switch event is directed by an increased 'accessibility' of the germline gamma 2b locus to switch-recombination enzymatic machinery. Our findings support the hypothesis that the apparently distinct recombination systems involved in variable region gene assembly and heavy chain class switching are both directed by the accessibility of their substrate gene segments. PMID- 3028779 TI - Novobiocin inhibits passive chromatin assembly in vitro. AB - Novobiocin, an inhibitor of prokaryotic DNA gyrase and eukaryotic type II topoisomerase enzymes, interferes with in vitro chromatin assembly using purified histones, DNA and nucleoplasmin. The target of inhibition is not topoisomerase II; this energy-independent assembly system lacks any ATP and Mg2+-dependent type II topoisomerase or gyrase activities. Rather, novobiocin interacts with histones, disrupting histone-histone associations required for octamer formation, and causing histones to precipitate from both nucleoplasmin-histone and histone DNA complexes. Thus, novobiocin is able to generate 'dynamic' chromatin in vitro in the absence of ATP and Mg2+ by removing histones from previously assembled static chromatin, so that the DNA supercoils, previously constrained by conventional nucleosomes, become susceptible to removal by topoisomerase I. PMID- 3028780 TI - Sequences sufficient for correct regulation of Sgs-3 lie close to or within the gene. AB - The Drosophila melanogaster 68C chromosomal locus is the site of a prominent polytene chromosome puff that harbors the genes Sgs-3, Sgs-7 and Sgs-8. These genes code for proteins that are part of the salivary glue that Drosophila larvae secrete as a means of fixing themselves to an external substrate for the duration of the pre-pupal and pupal period. The 68C glue genes are regulated by the steroid hormone ecdysterone, with the hormone required for both initiation and cessation of gene expression during the third larval instar. Previous work has defined sequences sufficient for expression of abundant levels of Sgs-3 mRNA at the correct time and in the correct tissue. We show here that sequences sufficient for normal tissue- and stage-specific accumulation of Sgs-3 RNA, but adequate only for low levels of expression, lie within 130 bp of the 5' end of the gene, or within the gene. PMID- 3028782 TI - DNA sequences required for expression of a Dictyostelium actin gene. AB - A 2.8-kb fragment of 5' non-coding DNA from the Dictyostelium actin 15 gene has previously been shown to contain all of the cis-acting DNA sequence elements required for normal developmentally-regulated transcription of actin gene fusion RNAs when reintroduced into the genome by DNA-mediated transformation. Deletion analysis of this promoter fragment indicates that all of the necessary information is contained within a 270-bp fragment of actin 15 5' non-coding DNA. This fragment contains four short G/C-rich repeated sequences that are also found in other co-regulated Dictyostelium actin genes. A 12-bp consensus sequence, AAAAATGGGG/ATT, is present in the regions essential for expression of two different Dictyostelium actin genes, actin 6 and actin 15, but is absent from an actin gene showing a different temporal pattern of developmental regulation. Deletion analysis and DNase I footprinting implicate this sequence as a functional cis-acting element required for transcription of the actin 15 gene. PMID- 3028781 TI - Evidence that white-blood is a novel type of temperature-sensitive mutation resulting from temperature-dependent effects of a transposon insertion on formation of white transcripts. AB - We report results indicating that the temperature-sensitive white-blood (wbl) mutation has a novel molecular basis, involving the formation of RNA transcripts of the affected gene rather than the behavior of the polypeptide product. First, we show that the temperature-sensitive mutant phenotype of wbl correlates with (and presumably results from) a temperature-dependent effect on levels of the 2.6 kb polyadenylated white transcript. Second, DNA sequence analysis and other studies show that wbl is associated with the insertion into the second white intron of a previously uncharacterized retrotransposon (designated the blood transposon). We discuss the potential origins of the novel temperature-sensitive molecular phenotype of wbl and the prospects for exploiting wbl to engineer temperature-sensitive mutations in other genes. PMID- 3028783 TI - Initiation of Escherichia coli minichromosome replication at oriC and at protein n' recognition sites. Two modes for initiating DNA synthesis in vitro. AB - The start sites for leading and lagging DNA strands were determined in vitro with minichromosomes as templates. Fragments from replication intermediates were analyzed by hybridization to single-stranded probes. Leading strand synthesis in the counterclockwise direction was found to originate in or close to (position 248 to -44) the minimal origin. Complementary lagging strand synthesis started several positions to the left outside of oriC. The results suggest in addition a concerted synthesis of leading and lagging strands following the dnaA directed assembly of initiation proteins at double-stranded oricC DNA (pre-replisome). In addition, DNA synthesis could initiate at protein n' recognition sequences located within and clockwise to the asnA gene. Initiation at n' sites was dependent on protein i activity, whereas leading and lagging strand initiation in the oriC region was not affected by protein i. Our results argue against an involvement of the phi X174-type primosome in the initiation of discontinuous DNA synthesis at oriC. An alternative function is suggested. PMID- 3028784 TI - On the mechanism of action of bovine intestinal mucosa 5'-nucleotide phosphodiesterase. Stereochemical evidence for a nucleotidyl-enzyme intermediate. AB - The diastereoisomers of adenosine 5'-O-phosphorothioate O-methyl ester have been synthesised. Only the Sp diastereoisomer is a substrate for the 5'-nucleotide phosphodiesterase from bovine intestinal mucosa. The previously unidentified enantiomer of 4-nitrophenyl phenyl phosphonothioate hydrolysed by the enzyme is shown to have the Sp configuration. Digestion of the Sp diastereoisomer of adenosine 5'-O-phosphorothioate O-methyl ester by the enzyme in 18O-labelled water gave 18O-labelled adenosine 5'-O-phosphorothioate which was stereochemically analysed by methylation and subsequent 31P-NMR spectroscopy and shown to possess the Sp configuration. Thus the enzyme-catalysed cleavage proceeded with retention of configuration at phosphorus, presumably via a double displacement mechanism. This provides strong evidence for the existence of a nucleotidyl-enzyme intermediate on the reaction pathway. PMID- 3028786 TI - Detection in situ of active polypeptides of mammalian and T4 DNA kinases after sodium dodecyl sulfate/polyacrylamide gel electrophoresis. AB - DNA kinase activity of rat liver nuclei was detected in situ after electrophoresis in sodium dodecyl sulfate/polyacrylamide gel containing 5' hydroxyl nicked DNA as DNA substrate. After renaturation of polypeptides, the gel was incubated with [gamma-32P]ATP and Mg2+. An active polypeptide corresponding to Mr 61,000 was observed as a radioactive band by autoradiography. The intensity of the band was proportional to the amount of the enzyme applied. The active band common to various tissues of rat was observed with the nuclear extracts, indicating that DNA kinase for rat tissue is composed of a single polypeptide of Mr 61,000. In contrast, T4 polynucleotide kinase (Mr = 140,000) showed an active polypeptide band corresponding to the subunit of Mr 33,000. PMID- 3028785 TI - Further biochemical characterization of an Na+ pump inhibitor purified from human urine. AB - An increase in endogenous Na+,K+-ATPase inhibitor(s) with digitalis-like properties has been reported in chronic renal insufficiency, in Na+-dependent experimental hypertension and in some essential hypertensive patients. The present study specifies some properties and some biochemical characteristics of a semipurified compound from human urine having digitalis-like properties. The urine-derived inhibitor (endalin) inhibits Na+,K+-ATPase activity and [3H] ouabain binding, and cross-reacts with anti-digoxin antibodies. The inhibitory effect on ATPases of endalin is higher on Na+,K+-ATPase than on Mg2+-ATPase and Ca2+-ATPase. The mechanism of endalin action on highly purified Na+,K+-ATPase was compared to that of ouabain and was similar in that it reversibly inhibited Na+,K+-ATPase activity; it inhibited Na+,K+-ATPase non-competitively with ATP; its inhibitory effect was facilitated by Na+; K+ decreased its inhibitory effect on Na+,K+-ATPase; it competitively inhibited ouabain binding to the enzyme; its binding was maximal in the presence of Mg2+ and Pi; it decreased the Na+ pump activity in human erythrocytes; it reduced serotonin uptake by human platelets; and it was diuretic and natriuretic in rat bioassay. The endalin differed from ouabain in only three aspects: its inhibitory effect was not really specific for Na+,K+-ATPase; its binding to the enzyme was undetectable in the presence of Mg2+ and ATP; it was not kaliuretic in rat bioassay. Endalin is a reversible and partial specific inhibitor of Na+,K+-ATPase, its Na+,K+-ATPase inhibition closely resembles that of ouabain and it could be considered as one of the natriuretic hormones. PMID- 3028788 TI - Chemical modification studies of beef-heart mitochondrial b-c1 complex. Effect of modification by ethoxyformic anhydride. AB - The effect of the histidine-modifier ethoxyformic anhydride (EFA) on the enzymatic properties of the mitochondrial b-c1 complex (ubiquinol-cytochrome c reductase) has been investigated. Chemical modification by EFA inhibited to the same extent the reductase and the proton translocating activity of the complex. In particular EFA modification of the complex resulted in: strong inhibition of the antimycin-insensitive reduction of b cytochromes; inhibition of the antimycin promoted oxidant-induced reduction of b cytochromes and inhibition of oxidation of pre-reduced b cytochromes. Analysis of the absorbance at 238 nm, indicative of N-(ethoxyformyl)histidine derivative, of the various polypeptide subunits separated by high-pressure liquid chromatography procedure, showed that EFA modified residues in core proteins and in the low-molecular-mass proteins. Both the inhibition of the redox and the protonmotive activity of the complex and the absorbance increase at 238 nm of the core protein fraction were readily reversed by hydroxylamine, indicating that modification of histidine residue(s) in core protein(s) is critical for the activity of the complex. This was supported by the finding that modification of the reductase with EFA prevented binding of fluorescein isothiocyanate to histidine residue(s) in core protein II. EFA modification of the reductase was without effect on the binding of N-(7 dimethylamino-4-methylcoumarinyl)maleimide to the various polypeptides of the complex except for the binding to the Fe-S protein which was greatly potentiated. Thus primary chemical modification of histidine residue(s) in core protein (II) appears to cause, in turn, a conformational change in the Rieske Fe-S protein. PMID- 3028787 TI - Role of cytosolic free calcium and phospholipase C in leukotriene-B4-stimulated secretion in human neutrophils. Comparison with the chemotactic peptide formyl methionyl-leucyl-phenylalanine. AB - Various leukotriene analogues were tested for their capacity to raise the cytosolic free calcium concentration, [Ca2+]i, and to stimulate exocytosis in human neutrophils. Their order of potency for both parameters was LTB4 greater than the stereochemical isomer of LTB4, (5S, 12S)-LTB4 much much greater than the sulphidopeptides LTD4, LTC4. The correlation between [Ca2+]i and secretion indicates that an increase of [Ca2+]i above a threshold level of about 300 nM is necessary for stimulating secretion with LTB4. This threshold is about an order of magnitude higher than that required for the chemotactic peptide formyl methionyl-leucyl-phenylalanine (fMet-Leu-Phe). The increase in [Ca2+]i elicited by LTB4 was unaffected by increasing cellular cAMP, while secretion was completely inhibited. These results indicate, that similar to fMet-Leu-Phe, leukotrienes generate other signals in addition to [Ca2+]i elevations. Contrary to previous claims, leukotrienes stimulate polyphosphoinositide hydrolysis, as indicated by the increase in [3H]inositol trisphosphate, InsP3, observed upon stimulation of myo[3H]inositol-labelled neutrophils with LTB4 or (5S, 12S)-LTB4. The two InsP3 isomers [Ins(1,4,5)P3 and Ins(1,3,4P3] were separated by high pressure liquid chromatographed and, as reported for other cell types, the formation of Ins(1,4,5)P3 precedes that of Ins(1,3,4)P3. Maximal stimulatory doses of LTB4 or (5S, 12S)-LTB4 produce about 50% the amount of InsP3 generated by equimolar concentrations of fMet-Leu-Phe. The present observations suggest that, though the transmembrane signalling systems activated by LTB4 and fMet-Leu Phe are the same, the different efficacy of these two agonists at stimulating neutrophil functions is due, at least in part, to a different degree of activation of phospholipase C. PMID- 3028789 TI - Purification and characterization of Desulfovibrio vulgaris (Hildenborough) hydrogenase expressed in Escherichia coli. AB - Hydrogenase from Desulfovibrio vulgaris (Hildenborough) is a heterologous dimer of molecular mass 46 + 13.5 kDa. Its two structural genes have been cloned on a 4664-base-pair fragment of known sequence in the vector pUC9. Expression of hydrogenase polypeptides in Escherichia coli transformed with this plasmid is poor (approximately 0.1% w/w of total protein). Deletion of up to 1.9 kb of insert DNA brings the gene encoding for the large subunit in close proximity to the lac promotor of pUC9 and results in a 50-fold increased expression of hydrogenase polypeptides in E. coli. The protein formed is inactive and was purified in order to delineate its defect. Complete purification was achieved with a procedure similar to that used for the isolation of active hydrogenase from D. vulgaris H. The derived protein is also an alpha beta dimer and electron paramagnetic resonance studies indicate the presence of the electron-transferring ferredoxin-type iron-sulfur clusters. In contrast to the native protein from D. vulgaris H, these can only be reduced with dithionite, not with hydrogen, indicating that the hydrogen-binding active centre which also contains an iron sulfur cluster is missing. PMID- 3028790 TI - Conformational changes in the oligonucleotide duplex d(GCGTTGCG) x d(CGCAACGC) induced by formation of a cis-syn thymine dimer. A two-dimensional NMR study. AB - In order to obtain insight into the repair mechanism of DNA containing thymine photo-dimer, the conformation of the duplex d(GCGTTGCG) x d(CGCAACGC) with a thymine dimer incorporated has been studied by proton NMR and the results are compared with NMR data of the parent octamer. Two-dimensional nuclear Overhauser enhancement (2D NOE) spectroscopy and two-dimensional homonuclear Hartmann-Hahn spectroscopy have been applied to assign all the non-exchangeable base protons and most of the deoxyribose protons of both duplexes. From these experiments it is clear that indeed a cis-syn cyclobutane-type thymine photodimer is formed by the irradiation of this oligonucleotide with ultraviolet light. Comparison of 2D NOE spectra and the 1H chemical shifts of the damaged and the intact DNA duplexes reveals that formation of a thymine dimer induces small distortions of the B-DNA structure, the main conformational change occurring at the site of the thymine dimer. PMID- 3028791 TI - Characterisation of the metal-ion-GDP complex at the active sites of transforming and nontransforming p21 proteins by observation of the 17O-Mn superhyperfine coupling and by kinetic methods. AB - Kinetic studies on the interaction of three Ha-ras-encoded p21 proteins with GDP and MgGDP have yielded values for the association (10(6)-10(7) M-1 s-1) and dissociation (10(-3)-10(-5) s-1) rate constants at 0 degrees C. Dramatic differences in the rate constants were not observed for the three proteins. Under non-physiological conditions (absence of Mg2+), the rate constant for GDP release was an order of magnitude faster for the viral protein p21v than for the cellular form p21c or the T24 mutant p21t, but this was reduced to a factor of about 3 in the presence of Mg2+. In all cases, there was an increase of about one order of magnitude in the rate of GDP release on removing magnesium. The binding affinities ranged from 5.7 X 10(10) M-1 for p21c to 1.3 X 10(11) M-1 for p21v. Electron paramagnetic resonance (EPR) measurements on Mn2+ bound together with stereospecifically 17O-labelled GDP showed direct coordination of a beta phosphate oxygen to the metal ion with a superhyperfine coupling constant of 0.16 0.22 mT, but no interaction with the alpha-phosphate oxygens at the active site of all three proteins. The association constant of Mn(II) to p21 proteins in the absence of nucleotides was estimated to be greater than 10(5) M-1. In agreement with the EPR results, experiments on the metal ion dependence of the binding of thiophosphate analogs of GDP provided further evidence for the absence of direct coordination of the metal ion to the alpha-phosphate group. These results have been used to construct a model for the interactions of Mg X GDP with the active site of p21 proteins. PMID- 3028792 TI - Isolation and characterization of a yeast mutant defective in cholinephosphotransferase. AB - A yeast mutant defective in cholinephosphotransferase (cpt) was isolated as a revertant from a choline-sensitive mutant, which exhibited lowered phosphatidylinositol synthesis. A block at the cholinephosphotransferase step in the mutant was indicated by the enzyme defect and the accumulation of CDP-choline in the cells with a decrease in phosphatidylcholine synthesis. The defect was due to a single recessive mutation in a nuclear gene. The residual activity in the mutant showed an increased apparent Km for CDP-choline and an altered sensitivity to Tween 20. Thus the structural gene may be affected in the mutant. The occurrence of an intact ethanolaminephosphotransferase in the mutant indicates the distinctness of the genes encoding cholinephosphotransferase and ethanolaminephosphotransferase in yeast. The present selection method was also effective for isolating mutants defective in the other steps of the CDP-choline pathway and choline transport. PMID- 3028793 TI - Modulation of human neutrophil function by C-reactive protein. AB - Investigation of the effect of C-reactive protein (CRP), an acute-phase reactant, on the functional capacities of human neutrophils was carried out as the basis for elucidating the biological function of C-reactive protein. An initial stimulation at low concentrations, followed by inhibition of superoxide production, and secretion of vitamin-B12-binding protein in the presence of two stimulants, phorbol myristate acetate and concanavalin A, and of neutrophil chemotaxis with increasing concentration of CRP was observed. Correlation between modulation of cell function, at least at relatively high CRP concentrations (greater than 50 micrograms/ml) and an increase in the intracellular level of cAMP is suggested. CRP was also found to enhance neutrophil phagocytosis of particles not containing phosphorylcholine, the native CRP ligand. The proposed role of CRP in neutrophil function is one of regulation and as a negative feedback for potential cytotoxic neutrophil functions. PMID- 3028794 TI - Stimulatory and inhibitory effects of protein kinase C activation and calcium ionophore on cultured pig Leydig cells. AB - The acute and the long-term (24 h) effects of protein kinase C activators, phorbol 12 myristate 13-acetate (PMA) and 1-oleoyl-2-acetyl-sn-glycerol, and the calcium ionophore A23187 on cultured pig Leydig cell functions were investigated. None of these drugs modified basal cAMP production, but they induced a small (3-4 fold) increase in testosterone secretion. The stimulatory effects of human choriogonadotropin (hCG; 1 nM) on both cAMP and testosterone productions were inhibited by short-term incubation with these drugs. In addition, they suppressed the stimulation of testosterone output by forskolin and 8-bromo-adenosine 3',5' monophosphate, whereas the forskolin-dependent cAMP production was unaffected. The inhibitory effects of PMA on hCG stimulation of both cAMP and testosterone were due mainly to a decrease of the Vmax without modification of the ED50. Moreover, PMA did not modify the binding of 125I-hCG. Pretreatment of Leydig cells with the three drugs for 24 h induced more pronounced modifications, such as a reduction in the number of hCG binding sites and a decreased responsiveness to hCG and forskolin, the testosterone production being drastically reduced. The effects of PMA were dose- and time-dependent; however, the concentration of PMA required to induce half-maximal effects on hCG receptors (10 nM) was about one order of magnitude higher than those required to reduce cAMP and testosterone productions. Further, the inhibitory effects on cAMP and testosterone secretions appeared within the first 3 h, whereas the hCG receptor number remained constant for at least 8 h. It appears therefore, that the main alteration responsible for the steroidogenic refractoriness of PMA-treated Leydig cells is located beyond cAMP formation. Moreover, since conversion of exogenous pregnenolone to testosterone by control and PMA-treated cells was similar, the alteration was probably located before pregnenolone formation. Kinetic studies with 125I-hCG showed that the rate of internalization of the hormone-receptor complexes was similar in control cells and in PMA-treated cells, suggesting that the decline in receptor number observed in the latter group after an 8-h delay is not due to an increased rate of internalization nor to sequestration of the internalized receptors inside the cells. Since cycloheximide blocked the effects of PMA on hCG down-regulation, it is likely that the phorbol esters and 1-oleoyl-2-acetyl-sn glycerol induce the synthesis of some proteins which blocked the recycling of internalized receptors. A similar hypothesis has been put forward recently to explain the hCG-induced down regulation.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3028795 TI - Purification and biochemical characterization of the complete structure of a proteolytically modified beta-2-microglobulin with biological activity. AB - A modified form of beta-2-microglobulin (beta-2-m) has previously been described to be present in serum from patients suffering from autoimmune diseases, acquired immune deficiency syndrome and small-cell lung cancer [Plesner, T. and Wiik, A. (1979) Scand. J. Immunol. 9, 247-254; Bhalla et al. (1985) Clin. Chem. 31, 1411 1412; Nissen et al. (1984) Clin. Chim. Acta 141, 41-50]. In the present study we describe the purification and characterization of this modified human serum beta 2-m from patients with small-cell lung cancer. Purified urinary beta-2-m was added to the serum samples incubated at 20 degrees C for five days to obtain a higher yield of modified beta-2-m (m-beta-2-m). m-beta-2-m was then purified from serum by gel filtration followed by chromatofocusing of the fractions containing beta-2-m. m-beta-2-m was found to have an apparent molecular mass of 15 kDa and a pI of 5.3 when analyzed by sodium dodecyl sulphate/polyacrylamide gel electrophoresis and analytical isoelectric focusing respectively. Amino acid analysis of m-beta-2-m revealed that the protein is missing one lysine residue compared to the composition deduced from the cDNA sequence of beta-2-m. Amino acid sequence analysis showed that m-beta-2-m consists of two polypeptide chains produced by a proteolytic cleavage of beta-2-m in the disulphide loop. After reduction and alkylation of m-beta-2-m the two chains were separated by reverse phase high-pressure liquid chromatography. By amino acid sequencing, amino acid residues 1-56 and 59-99 were identified in the A and B chains respectively. By comparison of the amino acid composition of m-beta-2-m with the known sequence of beta-2-m it was possible to deduce the existence of a Ser-57 in the A chain. Thus proteolytic cleavage of beta-2-m in the intrachain disulphide loop releases the amino acid Lys-58, which results in a modified form of beta-2-m with a molecular mass of 11,620 Da as determined by amino acid analysis. PMID- 3028796 TI - Ab initio quantum mechanical calculations of the variation of the 1H and 13C nuclear magnetic shielding constants in proline as a function of the angle psi. AB - The variation of the nuclear magnetic shielding constant of the different protons and carbons of trans HCO-L-Pro-NH2 with the value of the angle psi is calculated by a non-empirical method for three conformations of the proline ring. The results concerning the CH protons show that the chemical shift of the alpha, beta and gamma endo hydrogens can vary by more than 1 ppm when psi goes from -30 degrees to 180 degrees. The theoretical variation of the chemical shift difference between alpha and gamma or beta and gamma carbons is found to be sensitive to the puckering of the proline ring. For the second of these differences the theoretical results are in agreement with Siemion's relation only for a limited range of molecular conformations. Additional calculations show that the variations of the proton shifts with the value of psi are due to the magnetic anisotropy of the proline carbonyl group and to the polarization of the CH bonds by the multipolar charge distribution carried by this carbonyl. The results are discussed in relation to experiment and the possibility of using 1H and 13C chemical shifts for the determination of the value of the torsion angle about the C alpha C' bond. PMID- 3028797 TI - Subunit II of yeast QH2:cytochrome-c oxidoreductase. Nucleotide sequence of the gene and features of the protein. AB - The nucleotide sequence of a 2.5 X 10(3)-base segment of yeast nuclear DNA, containing the structural gene for the 40-kDa subunit II of the ubiquinol:cytochrome-c oxidoreductase, has been determined. The region contains only one single reading frame of length sufficient to encode a protein of the size of subunit II. The mature protein is predicted to have a length of 352 amino acids, with a molecular mass of 38714 Da. It is predominantly hydrophilic, with an overall polarity of 45%. Comparison of the sequence of the reading frame with that derived from direct sequence analysis of the N terminus of the mature 40-kDa protein shows that subunit II is synthesized as a longer precursor and shows that the extension is N-terminal. The presequence is 16 amino acids long and it contains a number of positively charged residues and lacks acidic ones. It is also rich in neutral, polar amino acids. S1 nuclease protection analysis of DNA X RNA hybrids identifies two major and one minor transcript of the gene, whose 5' termini map approximately 55, 65 and 75 nucleotides upstream of the initiation codon. Sequences 5' of these termini lack obvious homology to the regulatory sequences of other imported mitochondrial proteins, whose synthesis is controlled by oxygen and by catabolite repression. A mutant lacking a functional subunit II gene has been constructed by a one-step gene-disruption procedure. This mutant grows only slowly on glycerol and still displays a low level of QH2: cytochrome-c oxidoreductase activity (approx. 5% of that of wild type). The implications of this finding for the possible role of subunit II in the complex are discussed. PMID- 3028798 TI - Further studies on the structure of the glycogen-bound form of protein phosphatase-1 from rabbit skeletal muscle. AB - We have reported previously that the glycogen-bound form of protein phosphatase-1 (termed PP-1G) is a heterodimer, composed of the 37-kDa catalytic (C) subunit complexed to a 103-kDa G-subunit that anchors the enzyme to glycogen [Stralfors, P., Hiraga, A. and Cohen, P. (1985) Eur. J. Biochem. 149, 295-303]. An antibody raised against a synthetic peptide corresponding to the phosphorylation site on the G-subunit was found to immunoprecipitate PP-1G specifically. No precipitation occurred if the antibody was preincubated with the synthetic peptide, or if PP-1G was replaced by either the isolated C-subunit or protein phosphatase-2A. The results confirm by a new and independent method that the G-subunit is complexed to the C-subunit, and that it is not a contaminant. The G-subunit is remarkably sensitive to proteolysis. At the final stage of purification, PP-1G was eluted as a broad peak of activity. The leading fractions contained the 37-kDa C-subunit and 103-kDa G-subunit, while the central and trailing fractions comprised the 37 kDa C-subunit plus a number of bands with molecular masses ranging over 40-80 kDa. The 40-80-kDa bands were phosphorylated by cyclic-AMP-dependent protein kinase and tryptic digestion generated the identical phosphopeptides obtained by trypsinisation of the 103-kDa G-subunit. Furthermore, antibody to the G-subunit immunoprecipitated protein phosphatase activity quantitatively in the leading, central and trailing fractions. The results demonstrate that the 40-80-kDa polypeptides are fragments of the G-subunit, and that fragments as small as 40 kDa retain the sites of phosphorylation as well as the ability to interact with the C-subunit and with glycogen. Khatra [J. Biol. Chem. (1986) 261, 8944-8952] reported that the glycogen-bound form of protein phosphatase-1 did not contain a G-subunit and that it was a dimer composed of two identical C-subunits. The present work has shown that this proposal is incorrect, and that proteolysis of the G-subunit to fragments that stain very poorly with Coomassie blue can explain why this error was made. PMID- 3028799 TI - Receptor-mediated endocytosis of apamin by liver cells. AB - The binding and uptake of the bee venom toxin apamin, by guinea-pig and rat liver were studied. Guinea-pig liver plasma membranes contain inhibitable, high affinity binding sites for [125I]monoiodoapamin: Kd = 12.6 +/- 0.8 pM (SE); Bmax = 4.2 +/- 0.2 fmol/mg protein. No binding sites for [125I]monoiodoapamin on rat liver plasma membranes were detected in agreement with the absence of a physiological response to the toxin by rat hepatocytes. [125I]Monoiodoapamin, injected into the portal vein of guinea-pigs, was recovered in an undegraded form in a liver endosome fraction. The uptake of [125I]monoiodoapamin by rat livers was less than 4% of that taken up by guinea-pig livers and there was little evidence of radiolabelled toxin appearing in isolated rat endocytic vesicles. Inhibitable, high-affinity binding sites for [125I]monoiodoapamin were also identified on isolated guinea-pig liver endosomal membranes; Kd = 10.6 +/- 3.3 pM; Bmax = 2.5 +/- 0.6 fmol/mg protein. No inhibitable apamin binding sites were detected on rat endosomal membranes. Plasma membranes and endosomal membranes isolated from guinea-pig liver showed a similar spectrum of polypeptides to that previously reported for plasma membranes and endosomal membranes isolated from rat liver. The enzymatic composition of guinea-pig endosomes was also similar to that previously reported for rat endosomes. The results indicate that apamin was internalised by receptor-mediated endocytosis by guinea-pig liver cells in an analogous manner to that already shown for a variety of endogenous ligands. PMID- 3028801 TI - Influence of N6-methylation of residue A(5) on the conformational behaviour of d(C-C-G-A-A-T-T-C-G-G) in solution studied by 1H-NMR spectroscopy. 2. The hairpin form. AB - The solution conformations of the oligonucleotides d(C-C-G-A-A-T-T-C-G-G) and d(C C-G-A-m6A-T-T-C-G-G) as a function of temperature and sample concentration were investigated by means of 1H-NMR spectroscopy. The NMR spectra revealed that, at certain combinations of temperature and low sample and salt concentration, both compounds exist as a B-DNA-type duplex slowly (on the 1H-NMR time scale) interconverting with a monomeric species. From chemical shift data and imino proton spectra, it is concluded that the monomeric species consists of a mixture of a hairpin form in rapid equilibrium with the random-coil form. The double helical stem of the hairpin is formed by the six terminal cytidine and guanine residues, whereas the four core residues, -A-(m6)A-T-T-, partake in the loop. Thermodynamic analysis of the chemical shift of the resonances of the monomeric species vs temperature profiles of the two decamers and mutual comparison of these profiles indicate the following: the influence of N6-methylation of residue A(5) upon the local structure of the hairpin must be small; methylation decreases the stability of the duplex relative to the monomeric species: the temperature at which the fraction duplex equals 0.5 was found to be 312 K for the parent compound and 305 K for the methylated decamer at 2 mM sample concentration; methylation does not significantly alter the stability of the hairpin form relative to the random coil form: the Tm of the hairp----n equilibrium random coil equilibrium is 308 K for the parent compound and 306 K for the methylated decamer. A higher fraction hairpin-like structure for the N6-methylated compound is observed under identical conditions of temperature and sample concentration: at 300 K, 2 mM sample concentration, the fraction hairpin form is 0.12 for d(C-C G-A-A-T-T-C-G-G) and 0.20 for d(C-C-G-A-m6A-T-T-C-G-G). This finding appears to be a consequence of the reduced stability of the methylated dimeric species relative to the monomeric species, and to depend upon the sodium-ion concentration: it becomes more pronounced under low-salt conditions. PMID- 3028800 TI - Influence of N6-methylation of residue A(5) on the conformational behaviour of d(C-C-G-A-A-T-T-C-G-G) in solution studied by 1H-NMR spectroscopy. 1. The duplex form. AB - One- and two-dimensional NMR studies at 300 MHz and 500 MHz were carried out on the two oligonucleotides d(C-C-G-A-A-T-T-C-G-G) and d(C-C-G-A-m6A-T-T-C-G-G) in aqueous solution. NMR spectra were observed at 10 mM sample concentration over the temperature range 273-368 K. Assignments are given of the base, H1', H2', H2", H3' and of some H4' resonances, based upon a combination of two-dimensional correlation spectra (COSY) and two-dimensional nuclear Overhauser effect spectra (NOESY); imino-proton resonances were assigned with the aid of a two-dimensional NOE experiment. Chemical shift vs temperature profiles were constructed in order to gain insight into the influence of N6-methylation of residue A(5) on the temperature-dependent conformational behaviour of the decamer and to determine thermodynamic parameters for the duplex-to-coil transition. The NOESY spectra, the imino-proton spectra and the shift profiles of the two compounds, under conditions where each forms a B-DNA-type duplex, are very similar. This is taken to indicate that the influence of N6-methylation of residue A(5) on the local structure of the duplex must be small. However, the temperature dependence of the (non-)exchangeable proton resonances of the two compounds reveals that methylation slows down the duplex-single-strand exchange. Furthermore, a thermodynamic analysis of the two compounds indicates that N6-methylation slightly decreases the stability of the duplex relative to the monomeric forms (Tm is reduced from 332 K down to 325 K at 10 mM sample concentration). Proton proton couplings were obtained by means of one-dimensional and two-dimensional NMR experiments and were used in a conformational analysis of the sugar ring of each residue of the two compounds in the duplex form. The analysis indicated that all sugar rings display conformational flexibility in the intact duplex: population S-type sugar conformation ranges from 70% to 100%. A more refined analysis of the sugar rings of the parent compound revealed a sequence-dependent variation of the sugar geometry. This variation does not follow well the trend predicted by the Calladine/Dickerson sigma 3-sum rule [Dickerson, R. E. (1983) J. Mol. Biol. 166, 419-441; Calladine, C. R. (1982) J. Mol. Biol. 161, 343-352]; moreover the actual variations appear to be smaller in solution than those expected on the basis of known X-ray structures. PMID- 3028802 TI - Self-association and protonation of adenosine 5'-monophosphate in comparison with its 2'- and 3'-analogues and tubercidin 5'-monophosphate (7-deaza-AMP). AB - The concentration dependence of the chemical shifts of the protons H-2, H-8 and H 1' for 2'-, 3'- and 5'-AMP2- and of the protons H-2, H-7, H-8 and H-1' for tubercidin 5'-monophosphate (= 7-deaza-AMP2-; TuMP2-) has been measured in D2O at 27 degrees C to elucidate the self-association of the nucleoside monophosphates (NMPs). The results are consistent with the isodesmic model of indefinite non cooperative stacking; the association constants for all four NMPs are very similar: K approximately 2 M-1. These 1H-NMR measurements and those on the dependence of the chemical shifts on the pD of the solutions indicate that the NMP2- species exist predominately in the anti conformation. Comparison of the shift data for 5'-TuMP and 5'-AMP shows that no hydrogen bonding between N-7 and PO3H- occurs; hence, the previously observed and confirmed 'wrongway' chemical shift [Martin, R. B. (1985) Acc. Chem. Res 18, 32] connected with the deprotonation of the -PO3H- group most probably results from the anisotropic properties of the phosphate group which is in the anti conformation close to N-7. From the dependence between the chemical shift and the pD of the solutions the acidity constants were calculated for the four protonated NMPs, and for adenosine and D-ribose 5'-monophosphate. The measurements also allow an estimation of the first acidity constant of H3(5'-AMP)+ (pKDD3(AMP) = 0.9 and pKHH3(AMP) = 0.4). The values for pKHH2(NMP) and pKHH(NMP) were also determined from potentiometric pH titrations in aqueous solution (I = 0.1 M, NaNO3; 25 degrees C). The agreement of the results obtained by the two methods is excellent. The position of the phosphate group at the ribose moiety and the presence of N-7 in the base moiety influence somewhat the acid-base properties of the mentioned NMPs. Measurements with 5'-AMP in 50% (v/v) aqueous dioxane show that lowering of the solvent polarity facilitates removal of the proton from the H+(N-1) site while the -PO2-3 group becomes more basic; this increases the pH range in which the monoprotonated H(5'-AMP)- species is stable and which is now also extended into the physiological pH region. Some consequences of this observation for biological systems are indicated. PMID- 3028803 TI - Inhibitors of protein kinase C block the alpha 1-adrenergic refractoriness induced by phorbol 12-myristate 13-acetate, vasopressin and angiotensin II. AB - Vasopressin and angiotensin II inhibited in a dose-dependent fashion the stimulation of ureagenesis induced by alpha 1-adrenergic activation in hepatocytes incubated in medium without calcium and containing 25 microM EGTA. Vasopressin was more potent than angiotensin II. The effect of different inhibitors of protein kinase C on the alpha 1-adrenergic blockade induced by the vasopressor peptides was tested. It was observed that N-(6-aminohexyl)-5-chloro-1 naphthalene sulfonamide (W-7), 4-aminoethyl-1-[2,3-bis(n-decloxyl)-n-propyl]-4 phenylpiperadin e dihydrochloride (CP-46,665-1); 8-(N,N-diethylamino)octyl-3,4, 5 trimethoxybenzoate (TMB-8), polymyxin B and 1-(5-isoquinolynsulfonyl)-2 methylpiperazine (H-7) block this effect of the vasopressor peptides in a dose dependent fashion. The active phorbol ester, phorbol 12-myristate 13-acetate (PMA), also inhibited the alpha 1-adrenergic stimulation of ureagenesis in these cells. The inhibitors of protein kinase also blocked the effect of phorbol esters but a preincubation with the inhibitors before the addition of PMA was required. alpha 1-Adrenergic activation of phosphatidylinositol labeling was also abolished by PMA; the inhibitors of protein kinase partially blocked this effect of PMA. In summary, our data indicate that inhibitors of protein kinase C can block the alpha 1-adrenergic refractoriness induced by active phorbol esters, vasopressin and angiotensin II. The data are consistent with an important role of protein kinase C in modulating the alpha 1-adrenergic responsiveness of hepatocytes. PMID- 3028804 TI - Examination of the structural requirements for proliferation of peroxisomes and mitochondria in mouse liver by hypolipidemic agents, with special emphasis on structural analogues of 2-ethylhexanoic acid. AB - We have found here that there are clear structural requirements for peroxisome proliferation (monitored as increases in carnitine acetyltransferase activity, cyanide-insensitive palmitoyl-CoA oxidation, catalase and increases in the protein designated PPA 80) in mouse liver. From the investigation of ten structural analogues of 2-ethylhexanoic acid, it could be concluded that the most effective proliferators all have an ethyl group as the substituent on carbon 2 of the main chain, which consists of six carbons. The further observation from this group of compounds that a charged group is required for effective proliferation leads us to speculate that such a group is involved in the molecular mechanism as well. Many, but not all, of the effective peroxisome proliferators in a second group of compounds contain a phenoxy group, often with a substituted alpha carbon. Interestingly, the 2,4-dichlorophenoxyacetic and 2,4,5 trichlorophenoxyacetic acids are both effective peroxisome proliferators, but the closely related p-chlorophenoxyacetic acid is inactive in this respect, indicating that the chlorine atom at position 2 must be essential to the process in these cases. The results presented here also indicate that the structural requirements for proliferation of mitochondria are similar to those for proliferation of peroxisomes. Certainly, the most effective peroxisome proliferators also cause large increases in 'mitochondrial' protein and cytochrome oxidase activity, i.e. there is an obvious qualitative correlation. PMID- 3028806 TI - Osteoscintigraphy and brachydactylia of the hand. AB - Few reports have addressed scintigraphic abnormalities in the configuration of the hands. The purpose of this report is to present a case of congenital brachydactylia of the hands and to emphasize both acquired and projectional scintigraphic findings of brachydactylia. PMID- 3028805 TI - The trans-tubular network of the hepatocyte Golgi apparatus is part of the secretory pathway. AB - We have recently demonstrated the presence of sialyltransferase and sialic acid in a trans-tubular network (TTN) continuous with trans Golgi apparatus cisternae of rat liver hepatocytes. Based on these findings, we concluded that this structure, which also exhibited thiamine pyrophosphatase and acid phosphatase activity, is an integral part of the Golgi apparatus and functions in sialylation. In the present study, by comparing the distribution of a major hepatocyte secretory product with that of sialyltransferase, we sought to determine whether the TTN is also part of the secretory pathway. Examination of adjacent serial thin sections labeled for albumin showed its presence throughout the TTN and simultaneously provided new details about the structural complexity of the TTN. Double-immunolabeling with protein A-gold allowed the direct demonstration of albumin throughout the sialyltransferase containing TTN. Additional double staining protocols (combination of preembedding enzyme cytochemistry with postembedding immunolabeling) revealed the presence of albumin in both the thiamine pyrophosphatase and acid phosphatase positive regions of the TTN. These data show that albumin, a nonglycosylated secretory protein, reaches the TTN where terminal glycosylation of glycoproteins occurs. Therefore, it appears that the TTN of rat hepatocytes which functions in terminal glycosylation is also part of the constitutive secretory pathway. PMID- 3028807 TI - A case of virus-associated haemophagocytic syndrome with hypoplastic anaemia. AB - A 4-year-old girl with virus-associated haemophagocytic syndrome is described, suffering from fever, jaundice, rectal bleeding and a loss of consciousness. Severe pancytopenia was a prominent finding. The bone marrow biopsy showed hypocellularity, and bone marrow aspiration revealed proliferation of histiocytes containing many erythrocytes and thrombocytes. The patient recovered within several weeks of supportive therapy. Viral studies demonstrated a recent infection with Epstein-Barr virus. PMID- 3028810 TI - Desquamative interstitial pneumonitis and alveolar lipoproteinosis: diagnostic difficulties and therapy problems with an infant. AB - A desquamative, interstitial pneumonitis was diagnosed histologically in a 9 month-old boy who first became ill at the age of 5 weeks. The desquamative interstitial pneumonitis was associated with an acquired cytomegalovirus (CMV) infection. Despite treatment with corticoids, acyclovir and artificial ventilation, the patient died of pulmonary insufficiency at the age of 15 months. The autopsy revealed an alveolar lipoproteinosis. PMID- 3028808 TI - Multi-system coxsackievirus B-6 infection with findings suggestive of diabetes mellitus. AB - A fatal case of Coxsackievirus B-6 (CBV-6) infection in a 4 1/2-year-old girl is reported. The disease was initially characterized by a severe meningoencephalitis and, successively, by the appearance of hyperglycaemia and glycosuria, concomitantly with complement-fixing-islet cell antibodies (CF-ICA) and ICA, diarrhoea, electrolyte disorders, arrhythmia and decrease of the IgG levels, suggesting a multi-system involvement. CBV-6 was identified by isolation from stool and cerebrospinal fluid and by detection of specific IgM antibodies. PMID- 3028811 TI - Diagnostic significance of specific IgA coproconversion in rotavirus infection. PMID- 3028809 TI - Cushing disease: successful preoperative lateralization of an ACTH-producing pituitary microadenoma by simultaneous bilateral inferior petrosal venous sinus sampling with corticotropin-releasing hormone stimulation. AB - A 5 10/12-year-old girl with clinical and laboratory signs of endogenous hypercortisolism had evidence of ACTH hypersecretion in a standardized dexamethasone suppression test but had surprisingly low plasma ACTH concentrations before and after ovine corticotropin releasing hormone (oCRH) stimulation. To establish the diagnosis of pituitary disease and to localize the suspected microadenoma, we performed bilateral simultaneous inferior sinus petrosus blood sampling under CRH stimulation (oCRH 1 microgram/kg as an i.v. bolus). No major side effects were noted. Inferior petrosal blood ACTH concentrations did not differ from peripheral blood under basal sampling conditions but were higher in the effluent of the left half of the pituitary 5 min after oCRH stimulation. During transsphenoidal exploration of the sella an ACTH-producing microadenoma was removed from the left portion of the anterior pituitary gland. Signs of hypercortisolism remitted after surgery. Simultaneous bilateral sinus petrosus blood sampling under oCRH stimulation is a useful lateralization procedure even with small ACTH-producing adenomas. This technique may help to avoid unsuccessful surgical exploration in children with Cushing disease. PMID- 3028813 TI - Assay of epithelial membrane antigen (EMA) in human serum by ELISA. AB - An enzyme linked immuno sorbent assay (ELISA) for an epithelial membrane antigen (EMA) was described. Possible cross reactions with various antigens were investigated. In sera, EMA has been found in all the subjects studied. In normal population, levels were in the range of 500 ng/ml +/- 125 (S.D.) A significant increase was observed at the end of pregnancy and during lactation. A large number of patients suffering various benign and cancerous diseases were studied. The elevated levels found in breast and pulmonary pathology indicated that this assay could be useful in the follow-up of patients suffering from these diseases. PMID- 3028812 TI - Hormonal changes during a spontaneous attack of hypokalemic periodic paralysis. AB - We studied the hormonal changes during a spontaneous attack of hypokalemic periodic paralysis in a 20-year-old man, before and after treatment with potassium chloride. During paralysis, we observed high circulating levels of insulin, epinephrine, norepinephrine, growth hormone, ACTH and cortisol, most likely reflecting a condition of stress. Normalization of all these hormones occurred with recovery. Plasma aldosterone concentrations were normal. The increased plasma levels of insulin, but also of catecholamines and growth hormone, created a condition promoting potassium uptake in muscle cells. We suggest that stress may play a role in the pathophysiology of the paralytic attacks in this disorder. PMID- 3028814 TI - Search for HSV DNA in genital, cerebral and labial tumors. AB - DNA sequences homologous to HSV-1 and HSV-2 DNA fragments were searched in 64 genital, 35 labial and 34 cerebral tumors. Southern blot transfers of tumor and control DNAs were hybridized in stringent conditions with 32P labelled probes from HSV-1 and HSV-2 cloned DNA fragments. Specific hybridization to HSV-2 BglII N fragment was observed in six (9.4%) genital tumors. Labial and cerebral tumors did not show hybridization to any of the probes used. The technique employed allowed the detection of 0.1 copies of viral fragments per diploid genome. PMID- 3028816 TI - Transdermal delivery of bupranolol: pharmacodynamics and beta-adrenoceptor occupancy. AB - Bupranolol is a non-selective beta-adrenoceptor antagonist with a Ki-value of 6 15 nmol/l (equivalent to 1.5-4 ng/ml in plasma) at beta 1- (rat salivary gland) and beta 2-adrenoceptors (rat reticulocytes) in receptor binding studies with 3H CGP 12177 in the presence of human plasma. After oral administration of 200 mg bupranolol to healthy volunteers, the maximal plasma concentration was observed within 1.2 h but it only reached a level close to the Ki-value. Elimination from plasma was rapid (t 1/2 = 2.0 h). Administration of 30 mg bupranolol in a transdermal delivery system (TTS) every 24 h to 6 healthy volunteers for 72 h yielded steady state plasma concentrations 4- to 5-times above the Ki-value as shown by in vitro inhibition of beta-adrenoceptor binding by plasma samples. The pharmacodynamic effect, measured as the reduction in exercise tachycardia, showed a stable inhibitory effect; antagonism of a bolus injection of isoprenaline indicated a 10- to 15-fold right shift of the dose-response curve during the observation period of 72 h. It is concluded that steady-state plasma concentrations and effect of the elsewise rapidly eliminated beta-blocker bupranolol can be achieved by a transdermal delivery system applied each day. PMID- 3028815 TI - Changes in haemodynamics and body fluid volume due to enalapril in patients with essential hypertension on chronic diuretic therapy. AB - In 12 patients with essential hypertension who remained hypertensive despite chronic chlorthalidone treatment, the effect of 2 weeks of additional therapy with the converting enzyme inhibitor (CEI) enalapril on blood pressure and body fluid volumes has been evaluated. The objective was to examine the influence of a diuretic-stimulated renin-angiotensin-aldosterone system (RAAS) on haemodynamics and body fluid volume. Mean arterial pressure (MAP -21%), total peripheral resistance index (TPRI -22%) and plasma aldosterone concentration (PAC -39%) were decreased, and plasma renin activity (PRA 660%) was increased. The average heart rate (HR), cardiac index (CI), plasma volume (PV), blood volume (BV), extracellular fluid volume (ECFV) and body weight (BW) remained unchanged. A negative correlation was found between the per cent changes in ECFV and PAC. Thus, body fluid volumes during chronic diuretic treatment are well preserved even when the RAAS with its sodium retaining properties is suppressed by CEI. Possible mechanisms are a volume (not angiotensin II) - dependent stimulation of aldosterone and a fall in blood pressure. PMID- 3028817 TI - Activity of esterases in plasma from Ghanaian and British subjects. AB - We have measured aspirin esterase, cholinesterase, paraoxonase, and phenylacetate esterase activities in samples of plasma from British and Ghanaian subjects. Aspirin esterase, paraoxonase, and phenylacetate esterase activities were significantly lower in Ghanaians compared with British subjects. However, cholinesterase activities were similar in Ghanaian and British plasma samples. The lower esterase activities in Ghanaian plasma samples may result in higher circulating concentrations and greater pharmacological effects of drugs such as aspirin. PMID- 3028819 TI - Evidence for an association between CD23 and the receptor for a low molecular weight B cell growth factor. AB - Low molecular weight B cell growth factor (BCGF) and a monoclonal antibody (MHM6) to the 45-kDa, B lineage-restricted, CD23 activation antigen (BLAST-2; EBVCS) were found to be indistinguishable in their biological effects. Individually, both augmented DNA synthesis in activated, but not resting, B lymphocytes while no additional enhancement resulted from using the two agonists in combination. Furthermore, by increasing the expression of Tac, both MHM6 and BCGF promoted activated B cells to respond more vigorously to the late addition of recombinant interleukin 2. The presence of BCGF during B cell activations was found to down regulate the expression of the CD23 antigen while the coating of activated cells with MHM6 antibody diminished their capacity to absorb BCGF activity. The findings demonstrate that CD23 and a low molecular weight BCGF deliver a comparable growth-promoting signal to activated B cells. A possible relationship between CD23 and the receptor for the low molecular weight BCGF is discussed. PMID- 3028818 TI - Early events in human B cell activation: metabolic pathways vary according to the first signal used. AB - The effects of the calcium channel blocking drug Verapamil and of palmitoyl carnitine (PTC), an inhibitor of protein-kinase C activity, on human B cell activation were measured. Both Verapamil and PTC inhibited the B cell proliferation induced by costimulation with anti-mu antibody and with 3 different growth factors: interleukin 2, 20-kDa B cell growth factor and 50-kDa B cell growth factor. Both uridine and thymidine incorporation induced by costimulation with ionomycin and phorbol 12-myristate 13-acetate (PMA) were inhibited by Verapamil and PTC. In contrast, B cell proliferation was resistant to Verapamil (while being still inhibited by PTC) in two situations: when B cells were costimulated with PMA and growth factors and when B cells previously activated in vitro (by anti-mu antibody or PMA) were stimulated with growth factors. These results confirm that the late stage (G1----S transition) of B cell activation is independent of Ca2+ entry. More importantly, they show that the initial events induced by anti-mu antibody and by PMA are based on different biochemical pathways: PMA would act on a subpopulation of B cells which has already received an early signal of activation in vivo. This emphasizes the functional and biochemical heterogeneity of the G0 stage among circulating B cells. PMID- 3028821 TI - Soluble immune complex triggering of a respiratory burst in macrophages: the role of complex aggregation at the phagocyte surface. AB - The capacity of small, soluble immune complexes (SIC) to rearrange into larger aggregates, through additional antibody-antigen bridge formation, on a receptor bearing cell surface has previously been shown to play an important role in enhancing the uptake and ingestion of SIC by macrophages and has been implicated in the efficient clearance of SIC in vivo. In this report, we extend the study to investigate the role of SIC aggregation in triggering a respiratory burst in macrophages. SIC were found to be more efficient trigger signals than nonaggregating, antigen-free IgG oligomers of similar average size in that they induced a more rapid respiratory burst response that was between 2- and 6-fold greater in magnitude. The implications of these findings are discussed. PMID- 3028820 TI - Tumor-promoting phorbol esters suppress receptor-stimulated inositol phospholipid degradation and Ca2+ mobilization in mouse lymphocytes. AB - Anti-immunoglobulin antibodies (anti-Ig) provoke the rapid breakdown of phosphatidylinositol bisphosphate (PIP2), elevation of cytoplasmic Ca2+ levels ([Ca2+]i) and activation of protein kinase C (PKC) in B lymphocytes. Tumor promoting phorbol esters, like phorbol myristate acetate, also activate PKC, but inhibit anti-Ig-induced B cell proliferation. To investigate the basis of the latter effect, we studied the influence of phorbol esters on PIP2 degradation and [Ca2+]i in murine B cells. The results show that PKC-activating phorbol esters cause marked inhibition of anti-Ig-stimulated PIP2 breakdown and Ca2+ mobilization. In addition, these agents inhibit concanavalin A-provoked Ca2+ influx, lower resting cytoplasmic Ca2+ levels and reduce ionophore-induced Ca2+ influx in B cells. Apparently, PKC stimulation causes feedback inhibition of receptor signalling, not only by suppressing PIP2 degradation, but also by exerting additional complex effects on the control of [Ca2+]i in B cells. It is, however, not clear how these findings relate to the anti-proliferative effects of phorbol esters on B cells. PMID- 3028822 TI - Induction of H-2-specific antibodies by injections of syngeneic Sendai virus coated cells. AB - The capacity of the B cell immunoglobulin receptor to recognize complexes of Sendai viral and H-2b antigens was investigated by studying the antibody response to injections of syngeneic Sendai virus-coated (SV+) spleen cells in C57BL/6 (B6) mice. Almost all mice produced alloreactive anti-H2 lymphocytotoxic antibodies. In contrast, such antibodies were found very exceptionally in mice injected with normal (SV-) cells or with Sendai virus (SV) only. The reaction pattern of the cytotoxic antibodies induced was variable and ranged from almost anti-private to widely cross-reactive serotypes. The results of reactions on H-2-congenic, recombinant and -mutant mouse strains, and of capping and immunoprecipitation experiments showed that the cytotoxic antibodies were directed against H-2 class I molecules. The anti-H-2 antibodies exhibited enhanced binding for SV+ target cells, but absorption experiments showed that this was not the result of cross reactions with cell surface Sendai viral determinants or with a molecular complex of H-2 plus SV. This conclusion was supported by the observation that syngeneic SV+ cells were not the predominant targets for the induced lymphocytotoxic antibodies. Our results do not support the existence of MHC-restricted antiviral antibodies, but show the induction of anti-class I H-2 alloantibodies by injections with syngeneic SV-coated cells. We present a model for regular induction of anti-H-2 antibodies without intentional alloimmunization. PMID- 3028823 TI - Effect of chronic imipramine or baclofen on GABA-B binding and cyclic AMP production in cerebral cortex. AB - Long-term (14 days) treatment of mice with the antidepressant drug imipramine increased the number of GABA-B receptors in the cerebral cortex and also led to an increase in the potentiation of norepinephrine-induced cAMP accumulation by baclofen (a GABA-B agonist) in cortical slices. Chronic baclofen treatment decreased both of these measures. These results suggest that previous evidence of increased GABA-B binding by antidepressants is coupled to a functional increase in adenylate cyclase activity, but that the mechanism responsible for this effect is not due simply to direct GABA-B receptor stimulation. PMID- 3028824 TI - Papaverine-induced positive inotropism with failure to increase cyclic AMP in rat atria. AB - Phosphodiesterase inhibition by papaverine is likely to play a minor role in the rat atrial inotropic response because non-significant changes in cyclic AMP were obtained. Isoprenaline however raised the nucleotide levels three-fold and up to seventeen-fold when papaverine was added in a similar preparation. A prominent effect of papaverine was to lengthen relaxation instead of shortening it as did isoprenaline. The results suggest different sites of action, although overlapping effects cannot be excluded. PMID- 3028825 TI - Phospholipase C contracts visceral smooth muscle. AB - Phospholipase C added into the muscle bath (0.2-1.5 units/ml) contracted the guinea-pig taenia coli in a concentration-dependent manner. The fully developed contraction appeared within second, like the contractile effect evoked by 5 microM acetylcholine. Pharmacochemical dissection (using phospholipase D, indomethacin, neomycin, Li+, phorbol ester and oleoyl-acetyl-glycerol) showed that the contractions induced by exogenous phospholipase C are mediated by inositol trisphosphate, not by diacylglycerol. Thus, direct evidence is provided for the key role of the phospholipase C/inositol-trisphosphate system in smooth muscle contraction. PMID- 3028827 TI - Enkephalins inhibit non-adrenergic, non-cholinergic neuromuscular transmission in the human colon. PMID- 3028826 TI - 5-HT2 receptor-stimulated inositol phosphate formation in rat aortic myocytes. AB - Serotonin (5-HT) stimulated inositol phosphate production in primary cultures of rat aortic myocytes via a 5-HT2 receptor. Agonists active at 5-HT2 receptors in other systems were also active here but the response to some agonists was potentiated by the hormone uptake blocker, cocaine HCl. Two 5-HT2 selective antagonists, ketanserin and spiperone, inhibited the serotonin-induced response while compounds selective for other 5-HT receptor subtypes did not. PMID- 3028828 TI - [3H]CPP, a new competitive ligand for NMDA receptors. PMID- 3028829 TI - Newly synthesized noradrenaline mediates the alpha 2-adrenoceptor inhibition of [3H]5-hydroxytryptamine release induced by beta-phenylethylamine in rat hippocampal slices. AB - In slices of the rat hippocampus, alpha 2-adrenoceptors located presynaptically on serotonergic nerve terminals modulate the electrically evoked calcium dependent release of [3H]serotonin [( 3H]5HT). We have investigated the effects of a naturally occurring trace amine, beta-phenylethylamine (beta-PEA), on noradrenergic transmission in the rat hippocampus. Under experimental conditions in which MAO of type B is inhibited by deprenyl-exposure to beta-PEA (0.1-10 microM) facilitates the spontaneous outflow of [3H]noradrenaline and inhibits the electrically evoked release of [3H]5HT. The inhibitory effect of beta-PEA (3 microM) on the evoked release of [3H]5HT was antagonized by the alpha 2 adrenoceptor antagonist idazoxan at 1 microM, and by pretreatment with alpha methyl-p-tyrosine (alpha-MpT, 300 mg/kg i.p., 2 h). The inhibition of tyrosine hydroxylase activity by alpha-MpT does not modify the inhibition of the evoked release of [3H]5HT by the alpha 2-adrenoceptor agonist, 6-fluoronoradrenaline, or the serotonin receptor agonist, 5-methoxytryptamine. Pretreatment with the neurotoxin DSP4 (50 mg/kg i.p., 10 days) markedly antagonized the inhibitory action of beta-PEA on [3H]5HT release. These results indicate that the noradrenaline-releasing action of beta-PEA inhibits the electrically evoked release of [3H]5HT through the activation of alpha 2-adrenoceptors. This inhibitory effect appears to be mediated exclusively through the release of newly synthesized noradrenaline, and does not involve the direct activation by beta-PEA of the inhibitory 5HT autoreceptors which modulate [3H]5HT release in the rat hippocampus. PMID- 3028830 TI - Further evaluation of the selectivity of a novel antihypertensive agent, SGB 1534, for peripheral alpha 1-adrenoceptors in the spinally anesthetized dog. AB - Experiments were designed to examine some characteristics of an orally active antihypertensive agent, SGB-1534 on alpha-adrenoceptors in spinally anesthetized dogs. In the saphenous arterial bed perfused by a constant pump volume, saphenous nerve stimulation and bolus applications of norepinephrine and phenylephrine into the artery-evoked frequency- or dose-dependent increases (i.e. vasoconstriction) in perfusion pressure. SGB-1534 and prazosin infused i.v. significantly reduced the vasoconstriction in response to saphenous nerve stimulation and the two agonists. In the saphenous arterial bed, alpha 1-adrenoceptor antagonist potency of SGB-1534 on a weight basis was approximately 30 times greater than that of prazosin. Unlike SGB-1534 and prazosin, yohimbine failed to inhibit the vasoconstriction induced by phenylephrine, but instead potentiated the vasoconstrictor response to saphenous nerve stimulation. The equieffective doses of methoxamine and B-HT 920 given i.v. produced sustained pressor responses. SGB 1534 and prazosin applied i.v. in a cumulative way reduced dose dependently the pressor response to methoxamine but not that to B-HT 920. When the doses that blunted the sustained pressor response to methoxamine by 50% were compared, the alpha 1-adrenoceptor antagonistic activity of SGB-1534 was nine times greater than that of prazosin. PMID- 3028831 TI - Dihydropyridine sites separate from junctional sacroplasmic reticulum in hypertension. AB - Distribution patterns on linear sucrose gradients for dihydropyridine (DHP) binding sites and Mr 300K polypeptides marker for junctional sarcoplasmic reticulum were determined in left ventricular microsomes of control and chronically pressure-overloaded cat hearts. Major peak of DHP sites corresponded in position and shape to distribution of Mr 300K polypeptide in controls but appeared at a lighter buoyant density in hypertrophy, similar to junctional plasma membrane (T-tubules) isolated from normal microsomes by French press treatment of dyad fraction. PMID- 3028832 TI - Alpha 2-adrenoceptor-mediated inhibition of nerve stimulation-evoked release of neuropeptide Y (NPY)-like immunoreactivity in the pithed guinea-pig. AB - Clonidine (10 micrograms X kg-1) reduced by almost 50% the increase in plasma neuropeptide (NPY)-like immunoreactivity (-LI) induced by preganglionic nerve stimulation at 8 Hz. This effect was reversed by yohimbine (1 mg X kg-1) which caused a three-fold increase of the plasma NPY-LI. Prazosin (1 mg X kg-1) had no such effect. Guanethidine (5 mg X kg-1) reduced the stimulation-evoked increase in plasma NPY-LI. It is concluded that the release of NPY-LI evoked by nerve stimulation from sympathetic nerve terminals is controlled by a presynaptic alpha 2-adrenoceptor-mediated mechanism. PMID- 3028833 TI - The effects of adenosine on receptor sensitivity in the rat vas deferens. AB - In the guinea-pig isolated vas deferens adenosine augments the contractile response to noradrenaline (NA). Adenosine also augments the contractile response to NA and phenylephrine in the prostatic portion of the transversely bisected rat vas deferens. However, adenosine appeared to neither augment nor diminish the responses to carbachol, 5HT or high K+. Repetitive exposure of the vas to various agonists induced a desensitisation specific to the particular agonist used. The rate of return of responsiveness (the rate of resensitization) was investigated and, in particular, the effect of adenosine on that rate. In the rat vas deferens the rate of resensitization of responses to NA was retarded by adenosine and this effect was abolished by 8-phenyltheophylline and by yohimbine, but not by atropine or propranolol. Neither 8-phenyltheophylline, yohimbine, propranolol nor atropine by themselves affected the rate of resensitization of the rat alpha adrenoceptor. Adenosine had no effect on the rate of resensitization of responses to phenylephrine, 5HT, carbachol or high K+. It is considered that the potentiation of catecholamine responses by adenosine involves solely alpha 1 receptors but that the retardation of resensitization may involve alpha 2 receptors which do not contribute directly to contraction. PMID- 3028834 TI - Potentiation of bradykinin-induced nociceptive response by arachidonate metabolites in dogs. AB - Bradykinin was injected retrogradely into a branch of the splenic artery of lightly anesthetized dogs, and the reflex hypertensive response was used as an indicator of the nociception. The reflex hypertensive response to low doses of bradykinin (less than 1 nmol), but not to high doses (3-5 nmol), was markedly suppressed by infusion of indomethacin (0.54 mumol/min) into the splenic artery. During indomethacin infusion, the reflex hypertensive response to low doses of bradykinin was potentiated dose dependently by the following arachidonic acid metabolites (ED50): prostaglandin (PG)I2 (2.9 nmol) greater than or equal to PGH2 (4.4) greater than PGE2 (14) = thromboxane (TX)A2(15) greater than PGD2 (greater than 100). Leukotrienes B4, C4 and D4 showed practically no activity. This potentiating effect lasted up to 20 min after injection, particularly with PGE2. These results may not exclude the role of PGI2 as a major candidate for involvement in bradykinin-induced nociception, although a selective TX synthetase inhibitor (OKY-046) showed a weak analgesic effect (only 1/30 that of indomethacin). PMID- 3028835 TI - Anticholinesterase activity and structure activity relationships of a new series of hemicholinium-3 analogs. AB - Piperidine derivatives of hemicholinium-3 were synthesized and included the following spacing groups between the bis-cationic heads: trans,trans bicyclohexyl, phenanthrene, naphthalene and biphenyl. Anticholinesterase activity was determined using rat striatal synaptosomal cholinesterase, bovine erythrocyte acetylcholinesterase and horse serum butyrylcholinesterase. Hemicholinium-3 has little anticholinesterase activity but when the choline moiety of hemicholinium-3 is replaced with selected heterocyclic amine ring systems active inhibitors can be obtained. Results in this communication identify several quaternary amines which are equipotent with physostigmine for inhibition of cholinesterase. Several tertiary amines were also found to be active. Optimal anticholinesterase activity of these piperidine derivatives appear to be related to the necessity of 14 A interatomic distance between the cationic heads as well as C = O substitution in the phenylethyl spacing moiety. Reduction of C = O to secondary alcohol or CH2 results in decreased activity. The anticholinesterase activity is not only related to internitrogen distance and C = O substitution but also structural planarity and positional isomerism of the quaternary cationic head. PMID- 3028836 TI - High affinity and low affinity ouabain binding sites in the rat heart. AB - Ventricular muscle of rat heart has two classes of receptors which are responsible for the positive inotropic effect of ouabain. Low affinity receptors are apparently related to Na+, K+-ATPase. To determine if high affinity receptors are also sarcolemmal Na+, K+-ATPase of muscle cells, their characteristics were examined. Binding of [3H]ouabain to the high affinity binding site required ATP in the presence of Mg2+ and Na+, was stimulated by Na+ in the presence of Mg2+ and ATP, and was inhibited by K+. Digoxin, digitoxin and cassaine all inhibited [3H]ouabain binding to the high affinity site. Cassaine was about an order of magnitude less potent than the glycosides. These results indicate similarities in high affinity ouabain binding sites in ventricular muscle of rat heart and Na+, K+-ATPase obtained from other sources. Destruction of sympathetic nerve terminals with 6-hydroxydopamine failed to affect the high affinity ouabain binding sites indicating that high affinity sites do not represent the Na+, K+-ATPase in sympathetic nerve terminals. Labeling of Na+, K+-ATPase from [gamma-32P]ATP indicates that high affinity ouabain binding sites account for 25% of the total enzyme molecules present in ventricular muscle of rat heart. PMID- 3028837 TI - Modulation by cyclic AMP of arachidonic acid-induced platelet desensitization. AB - We have previously demonstrated that arachidonic acid (AA) and the stable cyclic endoperoxide analogue (U46619) desensitize human platelets at a common site, which is sensitive to endoperoxides/thromboxane receptor antagonists. We now report on the influence of agents which evaluate intracellular levels of platelet adenosine 3',5'-cyclic monophosphate (cAMP) on AA- and U46619-induced platelet desensitization. Prostaglandin E1, prostacyclin, carbacyclin, forskolin or dibutyryl cAMP prevented platelet activation by and desensitization to AA and to U46619 under conditions where the formation of thromboxane B2 was not significantly modified. Inhibition of platelet activation (aggregation and secretion) required a lower increase of the cAMP content than was needed to inhibit desensitization, confirming previous findings that desensitization to and by AA or U46619 are independent from the platelet release reaction. Together, these results indicate that AA-induced desensitization can be modulated by the adenylate cyclase/cAMP system acting at a site distinct from the known mechanisms of Ca2+ sequestration. This site is shared by the AA metabolite responsible for desensitization and by U46619 and is related to their common platelet membrane receptor. PMID- 3028838 TI - Interaction of kappa receptor agonists with Ca2+ channel antagonists in the modulation of hypothermia. AB - Administration of the opiate U-50,488H (3-20 mg/kg s.c.), a selective kappa receptor agonist, produced a dose-dependent decrease of rectal temperature in rats. This hypothermic effect of U-50,488H was accompanied by an enhanced activity of Ca2+/Mg2+ ATPase in crude synaptosomal (P2) fractions obtained from hypothalamus but not from cortex or cerebellum. Mg2+ ATPase activity in these regions was not altered by U-50,488H (15 mg/kg s.c.). Naloxone (5 mg/kg) partially and MR2266 (5 mg/kg) completely reversed the temperature and enzyme changes. Pretreatment with the calcium channel blockers nimodipine (1 mg/kg s.c.), diltiazem (10 mg/kg s.c.) and verapamil (2.5 mg/kg s.c.) potentiated the hypothermic effect of U-50,488H as well as the stimulation of Ca2+/Mg2+ ATPase in hypothalamus. These observations suggest that kappa agonists may produce opiate receptor mediated hypothermia through changes in intracellular Ca2+ levels in the hypothalamus. PMID- 3028839 TI - Influence of Ca2+e on 5-HT2- and alpha 1-induced arterial contraction and phosphoinositide metabolism. AB - Serotonin and phenylephrine were found to induce contractile responses and inositol phosphate (IP) formation in isolated rat tail artery. Both processes displayed similar concentration dependence in the presence of 2.5 mM external calcium (Ca2+e) and both were respectively inhibited by either ketanserine or prazosin, depending on the agonist used. In the absence of Ca2+e the amines no longer produced a contractile effect. In addition, lack of Ca2+ caused a shift to the right in the dose-response curve for phenylephrine-induced IP formation whereas serotonin-induced IP formation was not affected by changes in Ca2+e. The results suggest that alpha 1- and 5-HT2-induced contractions are quantitatively related to phosphatidylinositol metabolism. Contraction, but not IP formation requires the presence of Ca2+e. Different effects of Ca2+e on phenylephrine- and serotonin-induced IP formation could be related to a differential Ca2+ effect on binding of alpha 1- or 5-HT2 agonists. PMID- 3028840 TI - Antagonism of the ATP component of sympathetic co-transmission in the rat vas deferens by AMP. AB - Field stimulation of isolated rat vas deferens preparations caused a twitch contraction followed by a slower phasic contraction. Exogenously applied ATP and noradrenaline caused rapid contractions and slowly developing contractions respectively. Pretreatment with phentolamine antagonised the phasic response to stimulation and the contraction to noradrenaline. Pretreatment with AMP antagonized the twitch response to stimulation and concentrations to ATP. The results support the view that ATP is released as a co-transmitter with noradrenaline in sympathetic nerves. PMID- 3028841 TI - 'Peripheral-type' benzodiazepine receptors in the kidney: regulation of radioligand binding by anions and DIDS. AB - The high density of 'peripheral-type' benzodiazepine receptors (PBR) in the thick ascending loop of Henle (TAL) of the kidney prompted an evaluation of the effects of anions and anion channel blockers on these sites. [3H]Ro 5-4864 binding to rat kidney membranes was inhibited by halides in a concentration dependent fashion. Significant differences were observed in the efficacies (i.e. maximal effect achieved) of these halides to inhibit [3H]Ro 5-4864 binding. The effects of halides to inhibit this binding was manifest as a reduction in the apparent affinity of this radioligand with no effect on the maximum number of binding sites. The concentration necessary to achieve half maximal inhibition was around the physiologic range for Cl- in all extracellular body fluids. A strong correlation (r = 0.95; P less than 0.001) was obtained between the permeability of anions relative to Cl- and their efficacies to inhibit [3H]Ro 5-4864 binding. DIDS (4,4'-diisothiocyanostilbene-2,2'-disulfonic acid), an ion transport blocker, inhibited [3H]Ro 5-4864 binding to a maximum of 80%. The presence of Na2SO4 enhanced the potency of DIDS up to five-fold giving an IC50 of 43 +/- 6 microM. In contrast, the binding of [3H]PK 11195 was unaffected by DIDS and only slightly inhibited by I-. These data demonstrate that characteristics of radioligand binding to peripheral-type benzodiazepine receptors in the kidney are regulated by anions and anion transport blockers and suggest that the PBR may play an important role in anion transport. Additionally, these data suggest that the sites at which [3H]Ro 5-4864 and [3H]PK 11195 bind on the PBR may be overlapping, but not identical. PMID- 3028842 TI - Differences between rodent and human cell lines in the amount of integrated DNA after transfection. AB - The suitability of Chinese hamster and human cell lines for DNA-mediated gene transformation was investigated with respect to two parameters: the average quantity of and the integrity of integrated exogenous DNA fragments. No large differences were observed between most cell lines concerning the extent of fragmentation of the transferred DNA molecules. By contrast, the average number of sequences stably incorporated by the human cells (four lines tested) was 20- to 100-fold lower than the average amount inserted in the five Chinese hamster lines investigated. The very low uptake exhibited by the human cells, ranging from less than 100 up to 500 kb, renders these cells less suitable for transfection with genomic DNA to isolate specific genes. PMID- 3028843 TI - Activation of amino acid uptake at fertilization in the sea urchin egg. Requirement for proton compartmentalization during cytosolic alkalosis. AB - The comparative importance of the release of intracellular ionic calcium, Na+/H+ exchange and cytosolic alkalosis as activator signals was studied on the development of amino acid uptake at fertilization in sea urchin eggs. We show that, once stimulated, the rate of valine uptake is greatly dependent upon intracellular pH. Suppression of the Na+/H+ exchange at the time of activation, by applying ionophore (A23187) in sodium-free artificial sea water (ONaASW), inhibits the development of valine influx. This cannot be restored by a further (30 min later) alkalosis by transferring eggs into sea water. Suppressing the alkalosis in the presence of Na+/H+ exchange at fertilization by simultaneous addition of acid into sea water results in activation of the amino acid carrier which exhibits an increased rate of transport as soon as the eggs are replaced in sea water at pH 8.0. The absence of alkalosis in eggs activated in ONaASW can be counterbalanced either by adding NH4Cl 10 mM or by transfer into ASW at pH 9.0 at activation. Ammonia-treated eggs absorbed amino acid as controls, whereas eggs in sea water at pH 9.0 failed to develop a valine uptake system, suggesting that ammonia can completely replace the effect of Na+/H+ exchange. Furthermore, addition of NH4Cl immediately before fertilization conceals the Na+/H+ exchange but stimulates valine uptake as in controls. These data suggest that: the occurrence of the intracellular calcium increase alone is not sufficient for the develpment of the amino acid transport system; cell alkalinization at fertilization derives from the cytoplasmic membrane-located Na+/H+ exchange and an inward movement of protons into a cortical acidic compartment, which is discussed. PMID- 3028844 TI - Rous sarcoma virus-transformed fibroblasts and cells of monocytic origin display a peculiar dot-like organization of cytoskeletal proteins involved in microfilament-membrane interactions. AB - By immunofluorescence and interference reflection microscopy (IRM) we found that F-actin and a group of cytoskeletal proteins involved in microfilament-membrane interaction, including vinculin, alpha-actinin, fimbrin and talin, are specifically organized in discrete dot-like structures corresponding to cell substratum contact sites in both monocytes and monocyte-derived cells such as macrophages and osteoclasts. These proteins have a precise topological distribution; vinculin and talin form a doughnut-like ring, while actin, fimbrin and alpha-actinin are organized in dots matching the rings. An identical dot-like organization of F-actin and associated cytoskeletal proteins was also detected in malignant fibroblasts transformed by Rous Sarcoma virus (RSV) but not in the corresponding untransformed cells in culture. It is concluded that RSV transformation induces fibroblasts to express a cytoskeletal organization and a pattern of adhesion that are normally found in cells of monocytic origin. We propose that the occurrence of this cytoskeletal organization in RSV-transformed fibroblasts and in monocyte-derived cells may reflect a common ability to migrate across anatomical boundaries. PMID- 3028845 TI - 5-Bromodeoxyuridine-dependent increase in sister chromatid exchange formation in Bloom's syndrome is associated with reduction in topoisomerase II activity. AB - Bloom's syndrome is characterized by a high sister chromatid exchange (SCE) frequency, the basis for which is not yet understood. Immunofluorescent detection of SCE formation in dermal fibroblasts was employed over a wide range of 5 bromodeoxyuridine (BrdU) substitution into template DNA to show that this SCE elevation reflects both an increased baseline SCE frequency and an exaggerated increment in SCE formation as BrdU substitution increases. The impact of BrdU on SCE formation in Bloom's syndrome is paralleled by its ability to reduce the activity in nuclear extracts of topoisomerase II, an enzyme important for DNA replication and interchange. The extractable topoisomerase II activity of Bloom's syndrome fibroblasts, as measured by unknotting of page P4 DNA, is much more strongly inhibited by cell growth in medium containing BrdU than is that of normal fibroblasts. The results are consistent with the hypothesis that much of the BrdU-dependent component of SCE formation in Bloom's syndrome may be mediated by an effect of BrdU substitution of template DNA on topoisomerase II activity. PMID- 3028846 TI - Rat lens prostaglandin biosynthesis during galactose-induced cataractogenesis. AB - The relationship between the development of galactose-induced cataractogenesis and rat lens microsomal prostaglandin (PG) biosynthesis was studied. Within 24 hr of the introduction of 50% galactose to the rat's diet, lens PGF2 alpha production fell dramatically to 31% of control. Following the initial depression of PGF2 alpha biosynthesis, the ability to generate PGF2 alpha in lens microsomes slightly recovered reaching 58- and 53% of controls at day 2 and day 5 on the sugar diet, respectively. Determination of microsomal PGF2 alpha biosynthesis at 9- and 21 days revealed a continued decline in PG synthesis with complete cessation of PGF2 alpha synthesis by day 36 (hypermature cataracts, +5). The decreased PGF2 alpha biosynthetic capacity was a result of decreased cyclo oxygenase activity since: PGE2 production demonstrated a similar time course for inhibition; and lens microsomes from control and galactose fed rats revealed no difference in the PGs produced from PGH2 endoperoxide. Neither galactose (1 mM) nor galactitol (1 mM), when added to control microsomal preparations inhibited PG biosynthesis, eliminating the possibility of a direct effect of the sugar, or its metabolite, on cyclo-oxygenase activity. While PG biosynthesis was rapidly inhibited by the galactose feeding no changes were observed in basal lens cyclic AMP levels measured during the first 5 days of the feedings. These results demonstrate that depressed PG biosynthesis is an early consequence of galactose feeding. PMID- 3028847 TI - Human and experimental lens repair and calcification. AB - Lens repair and calcification have been studied in an experimental rabbit model of anterior segment necrosis. Findings were compared with those in a human senile cataractous lens with subcapsular calcification. Rabbit lenses subjected to anterior segment ischemia underwent a repair process similar to that observed in perforating lens injuries. Cellular response included the formation of fibroblast like cells that covered epithelial defects of the anterior pole. These observations suggest that the lens epithelium can transform into fibroblast-like cells. The calcification process was a non-cell-induced, and the observed mineral was probably nucleating on organic molecules. Elemental analysis demonstrated that crystals contained calcium and phosphorus with a ratio of 2:1. The mineral was probably hydroxyapatite. Since morphological findings in rabbit lenses closely resemble those of the studied cataractous human lens, the rabbit model appears to simulate one type of lens calcification in senile cataract. PMID- 3028849 TI - Aging of the lungs: speculation on the role of oxidants and proteases. AB - In most regards, aging of the human lungs mimics the development of emphysema. Various lines of evidence suggest that oxidants and proteases play an important role in the pathogenesis of emphysema. The disease seems to arise through a complex interplay of genetic and environmental factors. This paper explores the possibility that normal aging of the lungs, as it has been defined by various cross-sectional and longitudinal studies, is part of a pathological continuum ending with frank emphysema. It is not clear if these are "normal" aging effects or the result of the accumulation of disease. PMID- 3028848 TI - Mechanisms of age-related alterations of hormone secretion and action. An overview of 30 years of progress. PMID- 3028850 TI - Morphological evidence of an altered bone marrow microenvironment in patients with acute nonlymphoblastic leukemia and myelodysplastic disorders. AB - Type IV nuclear bodies are classified as true intranuclear inclusions that ultrastructurally contain numerous densely packed 20- to 30-nm osmiophilic granules surrounded by a microfibrillar cortex. In the present study, we have found statistically significant ultrastructural differences in the frequency and size of type IV nuclear bodies in the in vitro bone marrow fibroblastic cells (FC) derived from eight nonleukemic subjects and 13 patients with acute nonlymphoblastic leukemia (ANLL) and myelodysplastic disorders (MDD). Patients with ANLL, MDD, and myelofibrosis, as a group, had four times as many type IV nuclear bodies as nonleukemic subjects. The mean frequency of type IV nuclear bodies for patients with ANLL and MDD was 2.68% +/- 3.27% as compared with 0.63% +/- 1.06% for the nonleukemic subjects (p less than 0.05). The mean maximum type IV nuclear body area of the ANLL and chronic myelomonocytic leukemia (CMML) patients as a group was 2.08 +/- 1.10 micron2, compared with a mean maximum area of 0.93 +/- 0.10 micron2 from nonleukemic subjects (p less than 0.05). The FC were otherwise morphologically indistinguishable and displayed the typical ultrastructural features of fibroblasts. These findings have provided the first morphologic evidence that supports the concept of an altered bone marrow microenvironment in patients with ANLL and MDD. Since type IV nuclear bodies are found in high frequency in virally infected tissues, our quantitative ultrastructural findings raise the possibility of a local viral infection that affects the bone marrow microenvironment of patients with ANLL and some MDD disorders. PMID- 3028851 TI - Leukotriene production by fresh human bone marrow cells: evidence of altered lipoxygenase activity in chronic myelocytic leukemia. AB - The metabolism of arachidonic acid through the lipoxygenase pathway was studied in suspensions of fresh human bone marrow cells from eight patients with chronic myelocytic leukemia (CML) and 10 normal controls. After the cells were incubated with the calcium ionophore A23187 and arachidonic acid, a technique including reverse- and straight-phase high-pressure liquid chromatography (HPLC) was employed to isolate and detect different lipoxygenase-mediated compounds. The detected compounds included leukotriene B4 (LTB4), with its two major nonenzymatic isomers 6-trans-LTB4 and 12-epi-6-trans-LTB4 5S,12S-DHETE, and the monohydroxy eicosatetraenoic acids 5-HETE, 12-HETE, and 15-HETE. The pattern of lipoxygenase-mediated products from the bone marrows was similar to that previously described from human peripheral blood. Of eight bone marrow samples from CML patients, five expressed values above 600 ng LTB4/10(8) nucleated cells, as compared to only one out of 10 controls. In contrast, the CML patients produced significantly lower amounts of both the double-dioxygenation product 5S,12S-DHETE (56.8 +/- 16.0 ng [mean +/- SE] versus 146.1 +/- 31.3 ng; p less than 0.05) and the monohydroxy acid 12-HETE (965 +/- 351 ng versus 4390 +/- 1801 ng; p less than 0.05), indicating a 12-lipoxygenase deficiency. The present results show that leukotrienes are formed by human bone marrow cells and further suggest the existence of altered lipoxygenase activity in CML. PMID- 3028852 TI - Effect of donor and culture age on the function of neutrophils harvested from long-term bone marrow culture. AB - Neutrophils harvested from the supernatants of long-term bone marrow cultures that were initiated from young and old mice were tested for their functional activity. Superoxide generation was measured, as was the secretion of the enzymes lysozyme, myeloperoxidase, and glucuronidase, both basally and following stimulation by phorbol myristate acetate. The function of the neutrophils harvested from cultures that were initiated from young mice was identical to that of cells derived from young mouse peritoneal cavities at the time of death. A minimal decline in function in culture-derived cells was seen over the 15-18 weeks of study. For each of the above measurements examined, neutrophils obtained from cultures initiated from old mice gave values significantly lower than those in neutrophils recovered from cultures initiated from young mice. For some parameters (superoxide generation, myeloperoxidase), the rate of decline in function was more rapid for neutrophils from old rather than young mouse cultures. For other measurements the rates of decline were equal. The results indicated adequate neutrophil function in long-term bone marrow culture for extended periods of time. They also demonstrated that the age-related diminution in neutrophil function observed in vivo persisted in in vitro culture. PMID- 3028853 TI - Late, but not early, asthmatic reactions induced by toluene-diisocyanate are associated with increased airway responsiveness to methacholine. AB - We investigated whether airway responsiveness to methacholine differs in subjects with a history of sensitization to TDI who develop immediate, dual, late, or no asthmatic reactions after exposure to TDI, and also the effect of a TDI inhalation challenge in asthmatic subjects with hyperreactive airways with no history of sensitization to TDI. We measured FEV-1 immediately before and after exposure to TDI (0.018 ppm; 5-30 min) and then hourly for 8 h and the provocative dose (mg) of methacholine that caused a decrease in FEV-1 of 20% (PD20 FEV-1). The results of the present study suggest that the bronchoconstrictor effect of isocyanates is specifically linked to exposure to TDI and subsequent sensitization, excluding a nonspecific irritant effect on the airways. Moreover, they suggest that the increase in airway responsiveness to methacholine associated with the late asthmatic reaction is linked to factors that cause the late component of the asthmatic reaction. PMID- 3028854 TI - Lung cancer in Northern India in relation to age, sex and smoking habits. AB - Primary lung cancer in Northern India was analysed in relation to age, sex and smoking history in 480 histologically diagnosed cases. The M:F ratio rose progressively up to the 51-60 years age group and was about the same thereafter. The M:F ratio was very high in smokers and was almost identical for all cell types (greater than 20:1). In non-smokers, the M:F ratio differed with the cell type. The smoker:non-smoker ratio differed significantly in the age groups below and above 40. In those up to 40 years of age, the predominant cell type was small cell carcinoma and the cell type had no significant association with smoking habit. Above 40 years of age, the squamous cell type was most common in smokers and adenocarcinoma in non-smokers. PMID- 3028855 TI - Regional lung function in long-term survivors after chemotherapy for small cell lung cancer. AB - In order to evaluate the regional lung function in patients with small cell lung cancer with long-term survival, we measured the regional lung function in 17 patients. We studied the patients with 81m-krypton ventilation scans and 99m technetium perfusion scans, as well as measurements of static lung volumes (VC, RV, TLC), flow volume indices (FEV1, FVC, MEF50), and carbon monoxide diffusing capacity (DLCO). We found both the ventilation and the perfusion of the affected lung to be slightly but significantly lower (P less than 0.05), while no obvious ventilation or perfusion defects could be identified at the former tumor site. The static volumes of the lungs, the FVC and the FEV1 were within predicted normal values, while a significant decrease was seen in the PEF, MEF50, and DLCO. It is concluded that patients with small cell lung cancer who obtain complete remission with normalization of the chest X-ray and long-term survival after 18 months of intensive chemotherapy also have a nearly normal lung function. PMID- 3028856 TI - Return to work among patients with small cell lung cancer. AB - Forty-four patients with small cell lung cancer (SCLC), who were working in some kind of profession when they were taken ill, were studied with regard to return to work during or after cancer treatment. All patients received combination chemotherapy without chest radiotherapy or prophylactic brain irradiation. Twelve patients, referred to as W, returned to work during some period (14-83 weeks) after commencement of treatment, and 32 did not (NW). The stage of the disease prior to treatment and the type of profession seemed to be major prognostic factors. Patients with limited disease or "light" occupations more often returned to work than did patients with extensive disease or "heavy" occupations (P = 0.027 and 0.037, respectively). Age, sex and performance status were less important factors. Long-term survival was not a necessary condition for return to work, and overall survival was not prolonged in W patients compared with NW patients. It is concluded that increased occupational activity is an important palliative gain from chemotherapy in SCLC, and that this gain is not confined to a specific subgroup of patients, but is most likely to occur in patients with limited disease. PMID- 3028857 TI - Diffuse lung fibrosis due to fibrous zeolite (erionite) exposure. PMID- 3028859 TI - [Effect of gamma-aminobutyric acid analogs on the brain GABA-ergic system participating in cardiovascular regulation]. AB - In pentobarbital-anesthetized cats GABA methylester (MEG) and beta-phenyl-GABA methylester (MEF) administered intravenously in doses of 1/100, 1/30, 1/10 of LD50 and injected intracerebroventricularly in doses of 0.1 and 0.5 mg produce hypotension, These effects are significantly suppressed by picrotoxin. These compounds administered intracerebroventricularly in larger doses increase blood pressure, heart rate and sympathetic impulse outflow. Picrotoxin does not decrease these effects. MEG and MEF inhibit the GABA-transaminase activity of mouse brain in vivo. MEF increases and MEG decreases GABA concentration in the whole mouse brain. PMID- 3028858 TI - Ginkgo biloba extract inhibits oxygen species production generated by phorbol myristate acetate stimulated human leukocytes. AB - A Ginkgo biloba extract (Gbe) containing flavonoids, among other compounds, was tested for the release of activated oxygen species (O-2, H2O2, OH.) during the stimulation of human neutrophils (PMNs) by a soluble agonist. The extract slows down O2 consumption (respiratory burst) of stimulated cells by its inhibitory action on NADPH-oxidase, the enzyme responsible for the reduction of O2 to O-2. Consequently, superoxide anion (O-.2) and hydrogen peroxide (H2O2) production is significantly decreased when the PMNs stimulation is done in the presence of the extract at concentrations of 500, 250 and 125 micrograms/ml. Moreover, the hydroxyl radical generation (OH.) is very much decreased at concentrations as low as 15.6 micrograms Gbe/ml, which indicates that the extract also has free radical scavenging activity. Gbe is able at least to reduce very severely the activity of myeloperoxidase contained in neutrophils. This enzyme, secreted into the intra and extracellular medium, catalyzes the oxidation of chloride (Cl-) by H2O2 to yield strong oxidants (HOCl, chloramines) which are implicated in inflammatory processes. PMID- 3028860 TI - [Effect of trapidil and papaverine on thrombocyte aggregation and phosphodiesterase activity]. AB - Trapidil effect on platelet aggregation in vitro and in vivo as well as on phosphodiesterase (PDE) activity was studied. Both ADP- and collagen-induced aggregation of human and rabbit platelets was inhibited by trapidil at concentrations of 60 mumol/l. The PDE activity of human platelet lysates was blocked at concentrations that were necessary to inhibit aggregation. Papaverine exerted 5 times higher inhibitory effects on aggregation and PDE activity as compared to trapidil. Infusion of trapidil to rabbits caused aggregation inhibition in vivo. Papaverine at equimolar doses was more effective than trapidil. At therapeutic doses trapidil is unlikely to produce adequate plasma levels for inhibition of platelet aggregation. PMID- 3028861 TI - Synthesis of spin-labeled photoaffinity derivatives of NAD+ and their interaction with lactate dehydrogenase. AB - The synthesis of NAD+ derivatives spin-labeled at either N6 or C8 of the adenine ring is described, in which the carboxamide function of the nicotinamide moiety is replaced by a diazirine ring. Irradiation of these compounds at 350 nm generates a carbene which will react with any functional group in its vicinity including hydrocarbons. Both NAD+ derivatives form tight ternary complexes with lactate dehydrogenase and were covalently incorporated into this enzyme. They may be employed for ESR studies when non-covalent interactions are too weak for motionally restricted species to be observed. PMID- 3028862 TI - trp operon induction during the expression in E. coli of two IFN-gamma sequences. AB - Two nucleotide sequences coding for mature human immune interferon (IFN-gamma) and differing from each other by nine N-terminal nucleotides were expressed in E. coli under the control of a trp promoter. The longer gene variant after the ATG initiatory codon contained a TGT TAC TGC sequence, which was absent in the shorter gene. When expressed in E. coli under the direction of identical transcription and translation regulatory elements, these genes showed different susceptibility to induction. PMID- 3028863 TI - Calcium ionophore A23187 enhances human neutrophil superoxide release, stimulated by phorbol dibutyrate, by converting phorbol ester receptors from a low- to high affinity state. AB - The calcium ionophore A23187 acted synergistically with phorbol dibutyrate (PDBu) to stimulate human neutrophil superoxide production. A23187 shortened the lag period and markedly increased the initial rate of neutrophil superoxide production induced by suboptimal concentrations of PDBu. 1 microM A23187 reduced the EC50 value for superoxide release from 56 to 8 nM PDBu. This effect of A23187 was correlated with enhanced binding of [3H]PDBu to its receptor and a reduction in the dissociation constant (Kd) from 27 to 10 nM, without altering the apparent total number of phorbol dibutyrate receptors. These actions of A23187 were abolished in the presence of EGTA or TMB-8, confirming a dependence on Ca2+. PMID- 3028865 TI - Tetrameric alkaline phosphatase from human liver is converted to dimers by phosphatidylinositol phospholipase C. AB - Membrane-bound human liver alkaline phosphatase solubilized by a non-ionic detergent, Nonidet P-40 (NP-40), has the molecular mass of a tetramer. It can be converted to a dimeric form by treatment with a phosphatidylinositol phospholipase C (PI-PLC) obtained from Bacillus cereus. When human liver plasma membranes were directly treated with PI-PLC, the released alkaline phosphatase was dimeric. Thus, phosphatidylinositol may help maintain the tetrameric quaternary structure of alkaline phosphatase and aid its binding to human liver plasma membranes. PMID- 3028864 TI - The insulin receptor tyrosyl kinase phosphorylates holomeric forms of the guanine nucleotide regulatory proteins Gi and Go. AB - An affinity purified human insulin receptor preparation was shown to phosphorylate the alpha- and beta-subunits of the guanine nucleotide-regulatory proteins Gi and Go, derived from bovine brain. The presence of insulin stimulated the rate of their phosphorylation some 2-fold. The presence of Gi and Go did not affect the degree of autophosphorylation of the beta-subunit of the insulin receptor. Under conditions known to cause the dissociation of Gi and Go into their constituent subunits then phosphorylation of Gi and Go by the insulin receptor was abolished. The alpha-subunits of Gi and Go could be selectively phosphorylated by the insulin receptor tyrosyl kinase using appropriate concentrations of Mg2+ and GTP-gamma-S. PMID- 3028866 TI - The secretion of glucagon by transformed yeast strains. AB - Saccharomyces cerevisiae strains were transformed with plasmids coding for modified mating factor alpha 1 leader sequences followed by glucagon. Glucagon containing peptides which were secreted into the fermentation broth were isolated and their amino acid sequences determined. The yeast strain transformed with the sequence coding for the complete mating factor alpha 1 leader sequence preceding the glucagon gene (MT556) secreted glucagon plus glucagon extended at its N terminal by parts of the leader sequence. The yeast strain transformed with the sequence coding for a truncated mating factor alpha 1 leader sequence before the glucagon gene (MT615) secreted glucagon. These observations suggest that S. cerevisiae is a suitable vehicle for the efficient expression of plasmids coding for polypeptides similar to glucagon (e.g. VIP, secretin, GIP). PMID- 3028867 TI - Cardio-pulmonary elimination of 5-fluorouracil after bolus injection in the hepatic artery. AB - Theoretically, administration of 5-fluorouracil (5-FU) into the hepatic artery should improve the clinical response rate by high tumor cell extraction of the drug. However, previous studies in animals and man have also found a major extrahepatic elimination route for 5-FU. In the present study of 7 patients with primary and secondary liver cancer, we have found indications that a considerable elimination of 5-FU takes place in the cardio-pulmonary circuit. Thus, the mean AUC in the hepatic vein was 7185 mumol X min X l-1 and 3792 in a peripheral vein. PMID- 3028869 TI - Clomiphene citrate augments follicle-stimulating hormone-induced luteinizing hormone receptor content in cultured rat granulosa cells. AB - Although clomiphene citrate (CC) has been widely used for ovulation induction, the mechanism of its pro-fertility effect is not entirely clear. Because CC augments ovarian aromatase activity, the effect of CC on follicle-stimulating hormone (FSH)-induced luteinizing hormone (LH) receptor content was examined in primary cultures of rat granulosa cells. Control cultures contained low LH receptor content, whereas FSH stimulated LH receptor formation in a dose dependent manner. The concomitant addition of CC enhanced FSH potency as shown by a decrease in the ED50 for FSH by 2.2-fold. Some cells were treated with a low concentration of FSH (12 ng/ml) in the presence or absence of increasing concentrations of CC (10(-8) M to 10(-6) M). The enhancement of FSH-stimulated LH receptor formation was found to be CC dose dependent, with an ED50 of 6 X 10(-8) M. Thus, CC may play a direct ovarian role in the enhancement of FSH-stimulated LH receptors. The gonadal action of this compound may be relevant in understanding the efficacy of CC in ovulation induction. PMID- 3028868 TI - Independent occurrence of hepatocellular and cholangiocellular cancer nodules in the liver. AB - We treated a patient in whom hepatocellular and cholangiocellular cancer nodules in the liver were of independent occurrence. A 57-year-old Japanese male was admitted with a diagnosis of liver tumour, detected by a scan on a follow-up study of chronic liver disease. Computed tomography and hepatic angiography demonstrated two nodules located on different sites of the right lobe of the liver and which were suggestive of primary liver cancers. He underwent hepatic resection and each tumour was histologically proved to arise independently from hepatocellular and bile duct origins. This may be the first report of patient surgically treated for two different types of carcinoma in the liver. PMID- 3028871 TI - [Electrophysiological research on the excitation and inhibition evoked by ascending and corticofugal impulses in the relay neurons of the ventral posterolateral thalamic nucleus]. PMID- 3028870 TI - [Blocking of the GABA reception of the pyramidal neurons in isolated slices of the rat hippocampus]. PMID- 3028872 TI - [Characteristics of conditioned reflex excitation and inhibition in the neocortex]. PMID- 3028873 TI - [Neuronal electrical reactions of the 2nd auditory area of the cerebral cortex in cats to tones of a characteristic frequency]. PMID- 3028874 TI - [Effect of ACTH fragments on the aggressive-defensive behavior of the rat]. AB - ACTH4-10 and ACTH5-10 enhanced the fear response and footshock-induced aggression in male rats (50 and 150 mcg/kg, i.p.). ACTH4-10 seemed to produce the corticotropin (ACTH1-39) neuromodulatory behavioral effect which involved the facilitation of the brain muscarinic receptor activation. PMID- 3028876 TI - Activity of 5-nucleotidase and alkaline phosphatase in various organs and blood components of the pig. PMID- 3028875 TI - [Role of the gas composition of intraocular fluid in the regulation of intraocular circulation]. AB - The recovery velocity of the acido-alkaline state of the intraocular fluid depended upon the ocular hemodynamics in the rabbit intraocular chamber. Oppressive action of hyperoxia and substitute hypocapnia on intraocular blood circulation was revealed. The RQ reduction described in some eye diseases and after intraocular operations can be connected with changes in gaseous composition of the intraocular fluid. PMID- 3028877 TI - Histochemical studies of the intestine in mice with hymenolepiasis. PMID- 3028878 TI - [Basement membrane components in basalioma, cylindroma and squamous cell carcinoma]. PMID- 3028879 TI - Current aspects of research on the pathogenesis of diabetic neuropathy. AB - Diabetic neuropathy contributes to the long-term complications of diabetes mellitus. Its pathogenesis is still unknown although a host of abnormalities in peripheral nerves in human diabetics and animal models has been discovered. This article emphasizes the current metabolic hypotheses: increased polyol pathway, decreased myoinositol nerve content; altered phospholipid metabolism, decreased Na+/K+ ATPase activity and non-enzymatic glycosylation of proteins. Future perspectives relevant to animal and clinical research and new therapeutic aspects are also discussed. PMID- 3028880 TI - Effect of maternal food restriction on circulating insulin and glucagon levels and on liver insulin and glucagon binding sites of fetal and suckling rats. AB - Food was restricted in pregnant and nursing rats in order to evaluate the effect of malnutrition on insulin and glucagon metabolism in fetal and suckling rats. Food restriction of the mothers induced a loss of body and liver weights in their offspring, which was more pronounced in suckling than in fetal rats. A significant decrease of circulating insulin and glucagon levels in fetal and suckling rats from food restricted mothers (FRM) was also observed. In liver membranes insulin binding was higher in control, fetal and suckling rats than in adult animals, but maternal food restriction decreased insulin binding to liver membranes of suckling rats, and no changes between control and fetuses from FRM were observed. By contrast, glucagon binding was higher in adult than in younger control animals; however maternal food restriction had no effect on glucagon binding to liver membranes of fetal rats, although they produced an increase in 10 day-old suckling rats. The modifications in insulin and glucagon binding reflect changes in the number of receptors, but not in the affinity constants. The time courses of insulin and glucagon association to liver membranes were unaffected by the development or by the nutritional status of the animals. Degradation of insulin and glucagon by liver membranes was significantly lower in young rats and in rats from FRM, but this does not seem to be responsible for the differences observed in binding. No significant differences in the degradation of insulin and glucagon receptors between different groups of liver membranes were found.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3028881 TI - Carbon dioxide laser vaporization of cervical subclinical papillomaviral infection and intraepithelial neoplasia: short-term effectiveness. AB - 134 patients with biopsy-proved cervical subclinical papillomaviral infection (SPI) (the so-called 'flat condylomas'), or with associated intraepithelial neoplasia (also called 'atypical condyloma' or 'CIN with features of condyloma'), or with isolated, apparently 'non-warty' intraepithelial neoplasia were treated by means of the carbon dioxide laser between July, 1982, and January, 1985, following a planned approach and on an outpatient clinic basis, without the use of anesthesia or analgesia. The complication rate was extremely low. The overall cure rate was 91.0%. This confirms that laser surgery, combined with colposcopic expertise, can offer acceptable treatment effectiveness, comparable to other techniques of local destruction. Moreover, rapid tissue healing allows early identification of persistent disease. Converging cytological, histopathological, immunochemical and serological evidence has established human papillomavirus - belonging as a subgroup to the papovavirus family - as an important etiologic factor in female genital tract carcinogenesis. The marked increase in prevalence of these mainly sexually transmitted cervical SPI lesions creates a growing problem of their management and/or follow-up. PMID- 3028883 TI - In situ stability of uridylyl removing-uridylyltransferase of Escherichia coli. AB - The difference in sensitivity of UR/UT toward Lubrol WX permeabilization treatment of stationary phase E. coli cells is not uniquely related to nitrogen availability during cellular growth. The sensitivity of UR/UT to detergent treatment appears to be related to differences in the balance between fermentative and oxidative glucose metabolism. The possible occurrence of a third cycle in the glutamine synthetase regulatory cascade mechanism is considered. PMID- 3028882 TI - The effect of copper deficiency on heart microsomal phosphatidylcholine biosynthesis and concentration. AB - The effect of dietary copper deficiency on phosphatidylcholine biosynthetic enzymes, phosphatidylethanolamine methyltransferase, phosphatidyldimethyltransferase and choline phosphotransferase of heart microsomes was measured in rats. The data indicated that dietary copper deficiency can alter phosphatidylcholine biosynthesis and concentration in microsomal membranes of the heart. There was a significant decrease in the specific activity of choline phosphotransferase. There was a significant decrease in the concentration of total phospholipid-P, phosphatidylcholine-P, phosphatidylethanolamine-P, phosphatidylinositol-P, sphingomyelin-P and cardiolipin-P in the microsomes of the copper deficient animals. There was a significant decrease in the concentration of copper in microsomes of heart and liver in the copper deficient animals. PMID- 3028884 TI - Formation of HCN and its chlorination to ClCN by stimulated human neutrophils--2. Oxidation of thiocyanate as a source of HCN. AB - Leucocytes challenged by Staphylococcus epidermidis or stimulated by phorbol myristate acetate (PMA) produce cyanide from thiocyanate. The amount of H14CN formed depends on KS14CN concentration and is enhanced by pretreatment of phagocytosed bacteria with penicillin or by adding amine-taurine to the medium of PMA-stimulated neutrophils. The reaction of taurine chloramine or chlorinated Staphylococcus epidermidis (containing N-Cl groups) with thiocyanate results in HCN formation. At higher concentration of chloramine cyanogen chloride is formed. Cyanide is chlorinated by PMA-stimulated neutrophils and this process is significantly enhanced by exogenous taurine and inhibited by 3-amino 1,2,4 triazole. It is conceivable that oxidation of thiocyanate to HCN and chlorination of HCN to ClCN is mediated by the chlorinating species (taurine chloramine) produced by stimulated neutrophils. PMID- 3028885 TI - The deposition of calcium pyrophosphate by NTP pyrophosphohydrolase of matrix vesicles from fetal bovine epiphyseal cartilage. AB - About 5 mumol CaPPi/mg protein was deposited within 3 h in the presence of the reaction mixtures containing 1 mM ATP, 2 mM Ca2+, 1 mM Pi, and 17 micrograms of purified NTP pyrophosphohydrolase. At 1 mM ATP, 50% of the deposition was inhibited by 0.5-1 mM of various substrate and product analogues including AMP, ADP, and ethylene hydroxyl diphosphonate. The magnitude of inhibition on NTP pyrophosphohydrolase activity was in the order of AMP = CMP = ADP greater than adenosine greater than adenine greater than NAD = NADP. AMP, CMP, ADP, and adenosine are competitive inhibitors. The modes of inhibition by adenine, NAD, and NADP differ from the competitive inhibition. Ribose, 3'-AMP, 2'-AMP, and cAMP did not inhibit the enzyme activity. PMID- 3028887 TI - Regulation of amino acid transport in rat hepatocytes. PMID- 3028886 TI - Regulation of branched-chain amino acid metabolism. PMID- 3028888 TI - Nuclear events during early chondrogenesis: phosphorylation of the precartilage 35.5-kDa domain-specific chromatin protein and its regulation by cyclic AMP. AB - During chondrogenesis in vivo and in vitro, a family of nonhistone proteins (Mr 35,500), designated PCP 35.5, is lost from the nuclei of precartilage mesenchyme cells. A basic subcomponent of this family, designated PCP 35.5b, is phosphorylated during the first few hours of chondrogenesis in vitro by a phosphorylating system whose activity is enhanced 12- to 15-fold by exposure of differentiating precartilage cells to dibutyryl cyclic AMP. This phosphorylating system is present in isolated precartilage cell nuclei, where it retains its dependence on cyclic AMP and its specificity for PCP 35.5b. Assays for nuclear cyclic AMP inhibitable protein phosphatase activity capable of dephosphorylating PCP 35.5b were negative, indicating that the system responsible for phosphorylating this protein is a cyclic AMP-dependent protein kinase. Chromatin fractionation studies indicate that PCP 35.5b is localized at sites previously shown to be closely associated with DNase I-sensitive domains of precartilage cell chromatin. These studies define PCP 35.5b as a strategically located component of precartilage cell chromatin which is the major or sole chromatin target of cyclic AMP-dependent phosphorylation during chondrogenesis. This chromatin modification occurs prior to overt cartilage differentiation and may therefore play a regulatory role in the acquisition of the cartilage cell phenotype. PMID- 3028889 TI - Role of alpha 2-adrenergic receptor in platelet activation during insulin-induced hypoglycemia in normal subjects. AB - The effects of alpha 2-adrenergic-receptor blocker mianserin on the responses of blood glucose, plasma beta-thromboglobulin (beta-TG), and various counterregulatory hormones to insulin-induced hypoglycemia were studied in nine healthy male subjects. The alpha 2-adrenoceptor-blocking action of mianserin was confirmed by its inhibitory effect on platelet activation in vitro. Mianserin was given orally 90 min before insulin injection; the same study without mianserin was performed on another day as the control study. The time courses of blood glucose and serum C-peptide (0, 20, 45, and 180 min after the insulin injection) were identical in both studies, indicating that mianserin has no effect on these parameters. However, a significant increase of beta-TG at 45 min after insulin injection was completely suppressed by the administration of mianserin (mean +/- SE, 68.5 +/- 6.0 vs. 28.8 +/- 7.6 ng/ml, n = 6, P less than .05). No significant differences were obtained between the two studies in the responses of plasma or serum catecholamines, cortisol, glucagon, growth hormone, thromboxane B2, and 6 ketoprostaglandin F1 alpha. These results suggest that epinephrine is responsible for some, if not all, of the beta-TG release from the platelets during insulin induced hypoglycemia. PMID- 3028890 TI - Interactions of cholecystokinin and glucose in rat pancreatic islets. AB - The effects of sulfated cholecystokinin (CCK-8S) and glucose on insulin secretion and polyphosphoinositide (PPI) metabolism were studied in isolated rat islets. Both agonists stimulate PPI hydrolysis, inositol phosphate accumulation, 3H efflux from [3H]inositol-prelabeled tissue, and 45Ca efflux from prelabeled cells. However, the effects of CCK-8S on PPI metabolism are considerably greater than those of glucose. Furthermore, the effects of CCK-8S on PPI and Ca2+ metabolism are observed whether islets are incubated in either 2.75 or 7 mM glucose, but CCK-8S only stimulates insulin secretion (a biphasic response) when the higher glucose concentration is present. Addition of 1 microM forskolin to islets incubated in media containing 2.75 mM glucose does not influence basal insulin secretion but sensitizes the islets to the action of CCK-8S. In the presence of forskolin, CCK-8S induces a very marked first phase but no second phase of insulin secretion. We postulate that CCK-8S acts in this tissue via receptor-linked PPI hydrolysis, leading to an inositol trisphosphate-induced Ca2+ efflux. These receptor-mediated effects of CCK-8S are not altered either by the ambient glucose concentration or the cAMP content of the islets, but these two factors determine the responsiveness of the islets (in terms of insulin secretion) to a given CCK-8S signal. PMID- 3028891 TI - Polymorphic human glucose transporter gene (GLUT) is on chromosome 1p31.3----p35. AB - The glucose transporter is a membrane glycoprotein that is involved in the uptake of glucose by most, if not all, animal cells. A cloned cDNA that encodes the human protein was used to map the gene to a specific chromosomal region and to identify a DNA polymorphism. The human gene (designated GLUT) was assigned to chromosome 1p31.3----p35 by hybridization of the probe to DNA from a panel of human-mouse somatic cell hybrids containing different human chromosomes and by in situ hybridization to isolated metaphase chromosomes. The most likely location of GLUT is in 1p33. A common two-allele restriction-fragment-length polymorphism was identified with Xba I. PMID- 3028893 TI - An intrinsic neural pathway for long intestino-intestinal inhibitory reflexes. AB - We studied the mechanisms of initiation and pathways for the propagation of intestino-intestinal inhibitory reflexes induced by close intraarterial injections of neostigmine in conscious dogs. Two or three T-shaped catheters were surgically implanted in the intestinal branches of the superior mesenteric artery to inject pharmacologic agents locally in 10-15-cm-long segments. Migrating myoelectric complexes were recorded by a set of 10 electrodes and strain-gauge transducers. Close intraarterial injection of neostigmine initiated strong contractions of long duration in the perfused segment that terminated phase III activity in progress 90-150 cm distal or proximal to the cannulated sites and stopped its further migration. Atropine or 4-diphenylmethoxy-N-methylpiperidine methiodide injected just before neostigmine administration through the same catheter blocked both the local contractile effects and the reflex inhibition of phase III activity. Pirenzepine or hexamethonium injected in a similar manner did not affect the local response to neostigmine but blocked the reflex inhibition of phase III activity. A transection and reanastomosis in the mid-small intestine blocked the reflex inhibition by close intraarterial injection of neostigmine beyond the transection site. Pirenzepine, atropine, or hexamethonium injected through a middle catheter also blocked the reflex inhibition of phase III activity beyond the site perfused with these cholinergic antagonists. Close intraarterial administration of 4-diphenylmethoxy-N-methylpiperidine methiodide at a middle site had no effect on reflex inhibition. We concluded that strong spasmodic contractions in the small intestine initiate an intestino-intestinal inhibitory reflex in both directions. This reflex is mediated through an intrinsic neural pathway involving nicotinic and M1 muscarinic receptors. PMID- 3028892 TI - Noradrenaline and isoproterenol kinetics in diabetic patients with and without autonomic neuropathy. AB - Noradrenaline and isoproterenol kinetics using intravenous infusion of L-3H-NA and of 3H-isoproterenol were investigated in eight Type 1 (insulin-dependent) diabetic patients without neuropathy and in eight Type 1 diabetic patients with autonomic neuropathy matched for age, sex and duration of diabetes. Resting plasma noradrenaline and adrenaline concentrations were reduced in patients with autonomic failure (p less than 0.05). The metabolic clearance rate of noradrenaline was similar in both groups of patients, and the appearance rate of noradrenaline in plasma was reduced in patients with autonomic failure (p less than 0.01). The disappearance of L-3H-noradrenaline from plasma after the infusion of L-3H-noradrenaline had been stopped was not different in patients with and without neuropathy. The metabolic clearance of isoproterenol was not influenced by the presence of autonomic failure and mean values were similar to the corresponding values for noradrenaline. Isoproterenol was only taken up by a non-neuronal uptake; this finding may indicate that neuronal uptake is not important for the inactivation of circulating catecholamines. Alternatively, because the non-neuronal uptake of isoproterenol is probably greater than that of noradrenaline, we cannot exclude the possibility that a small decrease in the neuronal uptake of noradrenaline was compensated for by a slightly higher non neuronal uptake. PMID- 3028894 TI - Prostaglandin protects against taurocholate-induced damage to rat gastric mucosal cell culture. AB - Prostaglandins protect gastric mucosa against noxious agents, but it is unknown whether this protection includes a direct action on the cells themselves, this action is limited to damaging agents that inhibit prostaglandin synthesis, or cellular cyclic adenosine monophosphate is the mediator. The present study tested these questions in cultured gastric mucous epithelial cells. The effect of 16,16 dimethyl prostaglandin E2 on cellular cyclic adenosine monophosphate level and the effect of 16,16-dimethyl prostaglandin E2, dibutyryl cyclic adenosine monophosphate, and isobutyl methyl xanthine on taurocholate-induced damage to cultured rat gastric mucosal cells was determined. As parameters of cell damage, the trypan blue dye exclusion test and 51Cr-release were employed. Taurocholate significantly increased 51Cr-release in a dose-dependent manner and decreased the number of viable cells. 16,16-Dimethyl prostaglandin E2 (1.0 microM) diminished the cell damage caused by 10 mM taurocholate (p less than 0.01) and increased cyclic adenosine monophosphate levels. Prostaglandin F2 alpha but not prostaglandin I2 was also cytoprotective. Addition of dibutyryl cyclic adenosine monophosphate (1.0 mM) and isobutyl methyl xanthine while significantly increasing cyclic adenosine monophosphate levels did not significantly reduce taurocholate-induced cell damage. Thus, in vitro 16,16-dimethyl prostaglandin E2 directly protects gastric mucous cells against taurocholate-induced injury, direct prostaglandin cytoprotection is not limited to damaging agents that inhibit prostaglandin synthesis, and cyclic adenosine monophosphate levels do not correlate with gastric mucosal cell damage and may not be involved in the direct protective effect of prostaglandins. PMID- 3028895 TI - Emptying of the terminal ileum in intact humans. Influence of meal residue and ileal motility. AB - Emptying of the terminal ileum was assessed in 15 healthy humans by injecting technetium 99m-diethyltriaminopentaacetic acid into the bowel through a multilumen orocolonic tube. The subsequent arrival of isotope in the colon was quantified by gamma-scintigraphy and colonic filling curves were obtained. Studies were performed during fasting (n = 5) cnd 2.5 h after either a low residue meal (n = 5) or a meal made high in residue (n = 5) by adding 4 g of guar. The time for 50% of the isotope to reach the colon (T50) was significantly accelerated after both meals, being 72 +/- 15 min for the high residue meal and 62 +/- 8 min for the low residue meal, compared with 183 +/- 37 min (p less than 0.01) in the 5 fasting subjects. Although the addition of guar did not alter T50 significantly, it did cause a significant fall in the rate of colonic filling, implying increased isotope dilution. Delay at the ileocolonic junction, as shown by plateaus in the middle of the colonic filling curves, was uncommon. Hold-up was significant in only 2 of 10 postprandial and 2 of 5 fasting studies. Rates of ileocolonic transit could not be related to either a mean ileal motility index or the occurrence of specific ileal motor patterns immediately proximal to the ileocolonic junction. Fasting ileocolonic transit was characteristically erratic but could not be related to interdigestive migrating motor complexes, which were rarely observed in the last 60 cm of ileum. We conclude that ileocolonic transit in humans is related to the rate at which material accumulates in the ileum, being rapid postprandially (when ileal flow is high) and slow and erratic during fasting. This method yields consistent results and could be used to define further factors that influence ileocolonic inflow. PMID- 3028896 TI - Hepatic estrogen receptor in human liver disease. AB - Human liver contains estrogen receptors which render it sensitive to estrogen. Specific hormone binding to cytosol and nuclei from normal liver containing such receptors is of high affinity, low capacity, saturable, and specific for steroidal and nonsteroidal estrogens. Although estrogens alter metabolism and may produce disease, little data is available concerning estrogen receptor levels found in diseased liver. Herein we report estrogen receptor levels in human female liver containing diseases associated with oral contraceptives. Binding studies demonstrated cytosolic and nuclear estrogen receptors in human hepatic adenoma and focal nodular hyperplasia. Nuclear estrogen receptor levels in neoplastic tissue were greater than those in normal tissue. In addition, one hepatic adenoma resected from a patient taking tamoxifen contained no cytosolic estrogen receptor, and nuclear estrogen receptor levels were significantly lower than those found in normal tissue. These differences in binding capacity suggest a potential for greater hormone responsiveness in neoplastic liver tissue. PMID- 3028897 TI - Familial intestinal pseudoobstruction dominated by a progressive neurologic disease at a young age. AB - Chronic neuropathic intestinal pseudoobstruction is a rare entity, characterized by recurrent episodes of bowel obstruction without a mechanical obstructive cause. We report five members of two Jewish-Iranian families in whom chronic neuropathic intestinal pseudoobstruction was associated with an identical and unique progressive severe neuronal disease. It appeared within the first two decades of life. The disease consisted of external ophthalmoplegia, ptosis, and severe sensory and motor peripheral neuropathy. Three patients also had neuronal hearing loss. There was no evidence of central nervous system involvement and all patients were mentally intact. The combined disease was confirmed by radiologic, electrophysiologic, and histologic studies. Specific nutritional deficiencies, toxic elements, and systemic diseases affecting both the gastrointestinal tract and the nervous system were ruled out. It seems that these patients suffer from an autosomal recessive, presently unrecognized variant, of chronic neuropathic intestinal pseudoobstruction. In a patient with severe peripheral neuropathy of unknown etiology associated with symptoms suggestive of intestinal obstruction, the possibility of chronic neuropathic intestinal pseudoobstruction has to be considered. PMID- 3028898 TI - Comparative diagnostic value of the calcium-pentagastrin test versus the tolbutamide test in a patient with a somatostatinoma. AB - We describe a patient with a small somatostatinoma of the papilla of Vater without clinical evidence for diabetes mellitus, diarrhea, steatorrhea, or cholelithiasis, showing normal plasma basal levels for somatostatinlike immunoreactivity. The diagnosis was based on histologic and immunohistochemical analysis of tumor tissue and hypersomatostatinemia induced by the calcium pentagastrin test. Before removal of the tumor both diagnostic tests recommended for the detection of a somatostatinoma, a tolbutamide test and a calcium pentagastrin test, were performed. Whereas the calcium-pentagastrin test provoked a markedly elevated plasma somatostatin level in association with a depressed plasma neurotensin level, the tolbutamide test surprisingly did not. After removal of the tumor the calcium-pentagastrin test no longer induced hypersomatostatinemia. Further studies are needed to determine whether the calcium-pentagastrin test is a more reliable diagnostic test than the tolbutamide test in somatostatinomas with normal plasma basal levels. PMID- 3028899 TI - [Specific antibodies to herpes simplex virus, cytomegalovirus and rubella virus: distribution in amniotic fluid and blood]. AB - Fifty-nine unselected pregnant women were included in the study at the time of delivery. Immediately post partum maternal blood was obtained by venous puncture, and fetal blood from the umbilical cord. Amniotic fluid was obtained by amniocentesis during labor prior to rupture of the amniotic sac. Virus-specific rubella, herpes simplex and cytomegalo virus antibodies were determined with a commercial enzyme immune test. The virus-specific antibody titers are higher by a factor of 1.3 to 1.5 in the fetal than in the maternal serum. In contrast, the concentration of the corresponding antibodies in the amniotic fluid is approximately 5 titer levels (log 2) lower than in the maternal serum - regardless of the maternal antibody titer level. In view of its low concentration of antibodies the amniotic fluid can hardly be regarded as an "immunological barrier" if it serves as a "transport or distribution medium" in some virus infections. However, passive intra-amniotic immunization could be useful for combating certain virus infections during pregnancy, especially just before term. PMID- 3028901 TI - Modulation of somatostatin binding sites in cytosol of rabbit gastric fundic mucosa by cysteamine administration. AB - Cysteamine, when given in vivo to rabbits, depleted immunoreactive somatostatin in rabbit gastric fundic mucosa. Depletion of immunoreactive somatostatin was associated with both an increase in the number and a decrease in the affinity of the low-affinity somatostatin binding sites. The in vitro incubation of cysteamine (0.1 mM) with the cytosolic fraction did not result in any modification of somatostatin binding. These results suggest that a decrease in the endogenous immunoreactive somatostatin might lead to up-regulation of somatostatin binding sites in the gastric fundic mucosa. PMID- 3028900 TI - [Extramammary Paget's disease: clinical and histologic documentation of 2 forms]. AB - There are two forms of extramammary Paget's disease in respect of clinic, therapy and prognosis. In most cases (about 95%) extramammary Paget's disease is an intraepidermal manifestation of a cutaneous adenocarcinoma. After horizontal spreading within the epidermis, malignant growth continues along the adjacent adnexal structures with dermal spreading and subsequent metastases. In rare cases (approx. 5%), extramammary Paget's disease is an epidermal manifestation of an adenocarcinoma primarily located in the intestinal or urogenital tract. Further diagnostic and therapeutical procedures depend upon the type of extramammary Paget's disease. PMID- 3028902 TI - Actions of 4-(2-hydroxy-3-[(1-methyl-3-phenylpropyl)amino]propoxy) benzeneacetamide (KF4317) and labetalol on alpha- and beta-adrenoceptors. AB - Blocking activities of alpha- and beta-adrenoceptors by 4-(2-hydroxy-3-[(1-methyl 3-phenylpropyl)amino]propoxy)benzeneacetamide (KF4317) were tested on the isolated muscles, compared with the action of labetalol. In blocking the beta 1 adrenoceptor, KF4317 was almost as active as labetalol. KF4317 was about 1/10th as potent as labetalol in blocking the beta 2-receptor. KF4317 was beta 1 adrenoceptor selective. KF4317 was about 1/50th as potent as labetalol in blocking the alpha 1-receptor. KF4317 did not interact with the alpha 2 adrenoceptor in concentrations up to 10(-5) M. The present results indicate that KF4317 is a selective beta 1- and alpha 1-adrenoceptor blocker, though the alpha 1-adrenoceptor blocking action is weak. PMID- 3028903 TI - Iron binding to nutrients containing fiber and phenytoin. AB - Ascorbate and citrate extracted more iron than EDTA from pig foodstuffs (P less than 0.001). Glucose-saline solution (GSS) and ascorbate were less capable than citrate of extracting iron from maize tortilla (P less than 0.001). GSS extracted 16-32% of the whole iron contained in the studied foodstuffs; 30, 40 and 42% of the remaining iron in maize bran, wheat bran and pig feed was released with 0.1 M HCl. 40% of the remaining iron in pig's feed was released with 5% pepsin-0.1 M HCl while the iron of cereals required an additional extraction with 1% ascorbate. 5% pepsin-0.1 M HCl solutions released more iron as the protein content in the foodstuffs increased (P less than 0.001). Iron-extracted maize fibers (Ftw/Fe) bound significantly more iron than Ftc/Fe, FDA and FDN correspondingly (P less than 0.001). Phenytoin bound significantly more iron than total maize fibers both, with and without previous iron extraction (P less than 0.001). Also, phenytoin combined with maize-fiber bound less iron than phenytoin alone in G.S.A. PMID- 3028905 TI - The effect of dibutyryl cAMP on sodium uptake by isolated perfused gills of Callinectes sapidus. AB - Hemolymph, collected from 25 and 100% seawater acclimated Callinectes sapidus and perfused through the isolated gill, causes increases in Na+ uptake in the gill. Similarly, dibutyryl cAMP causes increases in Na+ uptake when perfused through the gill. Since pericardial organ (PO) extracts dopamine and octopamine are known to increase cAMP levels in the perfused gill, it is suggested that these substances from the PO may be the hemolymph factors which increase Na+ uptake via the mediation of cAMP as second messenger. PMID- 3028904 TI - Effects of cortisol and growth hormone on osmoregulation in pre- and desmoltified coho salmon (Oncorhynchus kisutch). AB - Salmonid species which undergo smoltification show a concurrent enhancement in saltwater (SW) osmoregulatory ability. This developmental change is marked by an increase in SW tolerance and gill Na+,K+-ATPase activity which appears to result, in part, from an increase in gill chloride cell density. Previous studies have suggested that cortisol and growth hormone (GH) may stimulate SW osmoregulatory mechanisms in salmonids. In this study, these hormones were examined for their ability to induce smoltification-associated osmoregulatory changes in pre- and desmoltified coho salmon (Oncorhynchus kisutch). Cortisol treatment for 12 days increased gill Na+,K+-ATPase activity in presmolts and gill residual (Na+,K+ independent) ATPase activity in both groups. Chloride cell density in presmolt primary and secondary lamellae and in desmolt secondary lamellae was increased as well. The rise in plasma sodium levels in fish transferred to SW was reduced only in desmolts. Treatment with bovine GH for 12-13 days increased gill Na+,K+-ATPase activity in presmolts and in desmolts. However, GH treatment in either group did not increase gill residual ATPase activity or alter plasma sodium levels in SW transferred animals. Gill chloride cell density in presmolts also was unaffected (desmolts were not examined). Thus, both cortisol and GH are partially able to produce changes similar to those observed during smoltification. The contrasting effects of these hormones on gill chloride cell density and gill residual ATPase activity suggest that cortisol may stimulate chloride cell proliferation and/or differentiation, whereas GH may act specifically to increase gill Na+,K+-ATPase activity. PMID- 3028906 TI - [Amplification of plasmid DNA inserted into the Bacillus subtilis chromosome]. AB - Amplification of plasmid pGG10 inserted into the Bacillus subtilis chromosome is described. The possibility of the 3.2 kb fragment of eucaryotic (wheat) DNA to be amplified within the bacterial genome is shown. The models explaining this phenomenon are discussed. PMID- 3028907 TI - [Detection of nucleotide sequences specific for retroviruses in Saccharomyces cells]. AB - Restriction endonuclease cleavage analysis and blotting hybridization of nuclear DNA and RNA to cloned avian sarcoma and murine leukemia virus genes (pol, scr and abl) demonstrated the presence and expression in baker's yeast cells of retrovirus-specific sequences. The relationship exists between the pol-specific yeast sequences and Ty cloned fragments. The results obtained are discussed in the light of evolutionary role of retroviral genes in cell division control and transposition. PMID- 3028908 TI - [Phenotypic manifestations of yeast transposon insertion in the LYS2 gene and deletions resulting from imprecise excision of the transposon]. AB - The lys2-32 mutant allele resulted from Ty1 element insertion was identified and cloned. The expression and reversions of lys2-32 localized in an autonomous plasmid were studied. The insertion was shown to inactivate LYS2 gene incompletely. Spontaneous reversions to complete or almost complete prototrophy were also obtained. About 50% of revertants retained the insertion. Others arise as a result of imprecise excision events leading to deletions of adjacent LYS2 sequences. PMID- 3028909 TI - [Expression of the quantitative trait radius incompletus, temperature effects and localization of mobile elements in Drosophila. I. Properties of test subpopulations]. AB - From the control sub-population, ric, with interrupted radial vein (L2) of the fly wing, two sub-populations were developed by selection: ris-, with distal and proximal fragments of L2 almost totally eliminated, as a result of minus selection; ris+, with totally restored radial vein, resulting from plus selection. Two sub-populations, ric113 and ric149, were also developed from the same original ric by changing gradually the cultivation temperature (29 degrees-- -18 degrees C) at the age of 113 +/- 5 h and 149 +/- 5 h, respectively. The former contained 2 times less and the latter 1.5 times more L2 than ric. These phenotypes were stably inherited in over 140 generations, expressing the "epigenic" properties. The genetic system of expression of ri oligogene was shown by genetic analysis to be corresponding to the polygene model of Mather. The main properties of 5 sub-populations used further for hybridization with mobile genetic elements, are described. Possible genetic mechanisms of the temperature effects are discussed. PMID- 3028911 TI - Genetic divergence between transposable elements. PMID- 3028910 TI - [Expression of the quantitative trait radius incompletus, temperature effects and localization of mobile genetic elements in Drosophila. II. Mobile genetic elements Dm-412]. AB - Two "selection" sub-populations (ris- and ris+), as well as two "temperature" ones (ric113 and ric149) were earlier developed from the control ric sub population with interrupted vein of the fly wing. All five sub-populations were investigated for hybridization of MGE Dm-412 with drosophila polytene chromosomes in situ. The tree of similarity of MGE Dm-412 hybridization patterns was built by the methods of matrix clusterization. The sub-populations with the most resembling expressions of characters (ris- and ric113, ris+ and ric149) were found to be also most similar in patterns of MGE localization and their changes. Nonrandomness of these changes was shown, the similarity of patterns being demonstrated to be mainly the result of the changes. There is evidence that such effects cannot be accounted for by genetic drift and independent stochastic changes in MGE localization. PMID- 3028912 TI - [Complex individual evaluation of the risk of developing diseases of the upper extremities as a result of overexertion by female finishers in the construction industry]. PMID- 3028913 TI - [Active infections with teratogenic viruses in parturients]. PMID- 3028914 TI - [Pathogenesis and diagnosis of disorders in the climacteric]. PMID- 3028915 TI - [A case of pleomorphic adenoma in the submandibular region]. PMID- 3028916 TI - Cyclic 3':5'-adenosine monophosphate phosphodiesterase and cyclic 3':5'-guanosine monophosphate phosphodiesterase of normal granulocytes and granulocytes of chronic myelogenous leukaemia. AB - Normal human neutrophilic granulocytes and granulocytes of chronic myelogenous leukaemia (CML) were shown to possess only the "high Km type" isoenzyme of cAMP phosphodiesterase. The properties of this enzyme are similar in both normal and CML granulocytes. cGMP-phosphodiesterase in human granulocytes was found to be composed of "high Km type" and "low Km type" isoenzymes. The high Km cGMP phosphodiesterase of CML granulocytes showed a considerably lower apparent Km and Vm value than those in normal neutrophilic granulocytes. PMID- 3028917 TI - Correlation of granulocyte intracellular activities of cyclic nucleotide phosphodiesterases with leukocyte count in patients with chronic myelogenous leukaemia. AB - The activity of adenosine cyclic 3':5'-monophosphate phosphodiesterase in granulocytes of patients with CML essentially depends on the granulocyte donor's WBC count. The ratio of cAMP-PDE/cGMP-PDE activities in CML granulocytes strongly correlates with CML host WBC count. The regression analysis of cyclic nucleotide phosphodiesterase activities and counts of individual constituents of the white blood cell population present in the blood of CML patients showed the primary relationship between the natural logarithm of total WBC count and the cAMP PDE/cGMP-PDE activity. The results suggest that the properties of CML granulocytes depend on the accumulation of these cells in the CML host. PMID- 3028918 TI - [Acute respiratory failure due to massive microembolization of the pulmonary arteries by breast tumor]. PMID- 3028919 TI - [131I antiferritin in the treatment of hepatoma and Hodgkin's disease]. PMID- 3028920 TI - [Rhabdomyolysis and damage to peripheral nerves due to drug intoxication]. PMID- 3028921 TI - [Studies on the diagnosis of insulinoma]. AB - The results of various tests for the diagnosis and localization of insulinoma in ten patients were reviewed. The diagnostic criteria IRI X 100/(BS-30)greater than 50 and IRI/BS greater than 0.30 in the fast were most reliable in establishing a diagnosis of insulinoma. If a diagnosis of insulinoma is in doubt after several overnight fasts, C-peptide suppression test should be carried out since the test is safe and convenient. As insulin stimulation tests often lead to false-negative results and sometimes cause dangerous levels of hypoglycemia in patients with insulinoma, they are not widely recommended now. However they may be useful, if positive, in some patients with insulinoma who exhibit no abnormality in insulin glucose ratio or C-peptide suppression test. Percutaneous transhepatic portal catheterization with measurements of radioimmunoreactive insulin concentration, supported by the clear theoretical grounds for interpretation of its results, was the most reliable method for preoperative localization of an insulinoma. Therefore the method, together with pancreatic angiography, should be attempted in all the patients with insulinoma. Insulinoma is an endocrine tumor curable by surgical removal, but there still remain a few unfortunate patients who have brain damages due to severe hypoglycemia or are blindly treated by pancreatectomy. Such cases should be eliminated by the early diagnosis and correct preoperative localization of the tumor. PMID- 3028922 TI - Effects of a kappa-opioid agonist on adrenocorticotropic and diuretic function in man. AB - Although kappa-opiate receptors represent an important fraction of the total opiate receptor capacity in human brain their endocrine function is unknown. We determined the effects of a kappa-opiate receptor agonist on the secretion of vasopressin, ACTH and cortisol and on diuresis. The racemic benzomorphan kappa agonist MR 2033 or its opiate active (-)-isomer, MR 2034, inhibited the release of cortisol and ACTH in 12 trials in a naloxone reversible manner; plasma levels of vasopressin were not altered. The (+)-isomer, MR 2035, did not affect the secretion of cortisol or ACTH. Surprisingly, in five other subjects large increases were observed in vasopressin, ACTH and cortisol following the kappa agonist, which were probably elicited indirectly by aversive effects of the opioid. The subjects in whom vasopressin release was not altered by MR 2033 and MR 2034 displayed large decreases in urine osmolality which were not antagonized by naloxone. The opiate inactive (+)-isomer, MR 2035, caused no diuretic response. Subjects in whom vasopressin release was stimulated did not show decreases in urine osmolality indicating that vasopressin is capable of antagonizing the diuretic action of the kappa-agonist. Our data show that a kappa agonist inhibits secretion of cortisol and ACTH by acting at stereospecific opiate receptors and elicits diuresis by acting at stereospecific, but naloxone insensitive non-classical opioid receptors. These data support the concept that different types of kappa-receptors can be distinguished in man. PMID- 3028923 TI - An ultrastructural study of mucoid carcinoma of the breast: variability of cytoplasmic features. AB - The ultrastructural characteristics of 14 cases of mucoid carcinoma of the breast, with different histological appearances, have been examined. Thirteen of the tumours were observed to consist of two populations of tumour cells, one showing secretory changes while the other group showed no evidence of activity. In one tumour only synthetically active cells were observed. Ultrastructurally, six different types of cytoplasmic granules, comprising typical mucin plus glycoprotein or protein-containing granules, were identified within the synthetically active tumour cells. The number of types of granules and the relative proportion of the various granules varied between tumours with only mucin granules present in all tumours. It would appear that synthetic pathways are activated in certain tumours which result in protein/glycoprotein granules associated with the argyrophilia observed histologically. In addition, the tumours varied with respect to luminal differentiation, presence of intracytoplasmic lumina, intracytoplasmic mucin pools, lipid droplets, ciliated cells and areas of calcification. The marked heterogeneity of the ultrastructural features of the mucoid carcinoma of the breast prevents the tumours from being readily divided into distinct subgroups. PMID- 3028924 TI - Proteinase inhibitors in the kidney and its tumours. AB - The proteinase inhibitors alpha-1-antitrypsin and alpha-1-antichymotrypsin are found in a variety of tissues including the kidney. We have used the immunoperoxidase technique to study the distribution of these glycoproteins in 10 fetal and five adult kidneys and in 44 renal tumours. Proteinase inhibitors were found in fetal kidneys in the cells of the metanephric blastema, primitive mesenchyme and in proximal tubules. They were found in proximal tubules of adult kidneys. These substances were also found in the mesenchyme of nephroblastomas and in the tumour cells of clear cell, oncocytic and sarcomatoid variants of renal cell carcinoma. These observations are significant in the understanding of the patterns of differentiation of which renal cell carcinoma is capable. PMID- 3028925 TI - Carney's triad: gastric leiomyosarcoma, pulmonary chondroma and extra-adrenal paraganglioma in young females. AB - The case of a 13-year-old girl with a gastric leiomyosarcoma and a pulmonary osteochondroma is presented. The association of these two tumours and extra adrenal paraganglioma has been described as a triad by Carney. The patient is free of recurrence of the gastric tumour with no evidence of paraganglioma 10 months after the operation. To our knowledge this is the first case of the triad reported in the UK. PMID- 3028926 TI - Intercellular junctions and tumor stage in small cell carcinoma of the lung. AB - The authors have studied the ultrastructural features of 52 cases of oat cell carcinoma of the lung and have related their observations to tumor stage and patient survival. Only the type of cell junctions seems to be of prognostic importance. Tumors with intermediate junctions--and especially those with desmosomes--have a more localized stage and may be resectable to result in longer survival than expected for oat cell carcinomas without junctions. For example, in the authors' series the median survival periods for those with no identifiable junctions, intermediate junctions, or desmosomes were 6.4, 8.2, and 11.3 months, respectively. Nevertheless, this ultrastructural subclassification is not as effective as that obtained from careful clinical staging. PMID- 3028927 TI - Neural differentiation in Wilms' tumor. AB - Immunoperoxidase studies using antibodies to neuron-specific enolase (NSE), glial fibrillary acidic protein (GFAP), and S100 were used to confirm the presence of neural differentiation identified histologically in five cases of Wilms' Tumor. These observations invite reappraisal of Masson's proposed neuroectodermal histogenesis of Wilms' tumor because they are difficult to reconcile with the currently accepted metanephrogenic blastematous origin. They also provide a stimulus to examine the expression of neural markers during normal renal development and differentiation. PMID- 3028928 TI - Complete and incomplete Drash syndrome: a clinicopathologic study of five cases of a dysontogenetic-neoplastic complex. AB - Drash syndrome is a complex disorder characterized by abnormal renal function, abnormal sexual differentiation with predisposition to developing gonadal neoplasms, and nephroblastoma. The authors report five cases with various manifestations of this syndrome. Dysgenetic gonads and abnormal sexual differentiation were present in all patients; two had unilateral and two bilateral gonadoblastomas; in addition, one of the latter had a juvenile granulosa cell tumor. Renal failure was present in all patients. One patient had bilateral Wilms' tumor, and one patient had a metanephric hamartoma. Each element of the triad in this syndrome is analyzed with regard to possible pathogenetic mechanisms and current models of carcinogenesis. Cases with complete forms of the syndrome reported in the literature are reviewed. Patients with incomplete forms of the syndrome must be followed carefully because other elements of this complex may become manifest. PMID- 3028929 TI - Partial gastrectomy: a risk factor for carcinoma of the pancreas? AB - Recent reports suggest an increased risk of cancer of the pancreas after partial gastrectomy. We investigated this putative relationship by performing a case control study on autopsy subjects. Our findings suggest a three-fold risk of pancreatic carcinoma in postgastrectomy patients. Increased production of carcinogens in the gastric remnant, excretion of the carcinogens by the liver into the bile, and subsequent bile reflux into the pancreatic duct is proposed as a working hypothesis for pancreatic carcinogenesis. This study illustrates the usefulness of autopsy series as an instrument for checking a straightforward hypothesis before designing more extensive and expensive studies. Moreover, efficient use of autopsy data was accomplished by computerized data base retrieval of autopsy records, emphasizing that this method of record retrieval is an important precondition for prospective studies. PMID- 3028931 TI - Pleomorphic rhabdomyosarcoma in adults: immunohistochemistry as a tool for its diagnosis. AB - Pleomorphic rhabdomyosarcoma in adults over 30 years of age was a diagnosis frequently made in the 1960s and 1970s. Since the general acceptance of malignant fibrous histiocytoma (MFH) as a tumor entity at the end of the 1970s, however, it has become a very rare tumor in adults. Therefore, 21 cases originally diagnosed on the basis of histology and clinical data as pleomorphic rhabdomyosarcoma in the 1960s and 1970s were reexamined immunohistochemically. Other types of pleomorphic sarcomas involved in the differential diagnosis were also studied. Specific antibodies against vimentin, desmin, creatine kinase subunit M, skeletal muscle actin and myosin, and myoglobin, and the avidin-biotin-peroxidase complex technique were used. The immunohistochemical findings indicate that rhabdomyosarcoma occurs only rarely in adults over 30 years of age and that the majority of the tumors have to be reclassified as MFH or leiomyosarcoma. On the other hand, several pleomorphic sarcomas were found to be diagnosed incorrectly as MFH or liposarcoma by routine histologic stains and electron microscopy. The revised diagnosis was pleomorphic rhabdomyosarcoma for one case and pleomorphic leiomyosarcoma for the other cases. Thus, this study clearly shows the usefulness of immunohistochemistry as a technique in the diagnosis of pleomorphic sarcomas in adults. PMID- 3028930 TI - Cytomegalovirus encephalitis in patients with acquired immunodeficiency syndrome: an autopsy study of 30 cases and a review of the literature. AB - The pathology of cytomegalovirus (CMV) encephalitis was studied at autopsy in thirty patients with acquired immunodeficiency syndrome. Lesions could be segregated into five major categories: microglial nodules, isolated inclusion bearing cells, focal parenchymal necrosis, necrotizing ventriculo-encephalitis, and necrotizing radiculo-myelitis. Microglial nodules and CMV inclusions were present in all brains. Microglial nodules were found with variable frequency and had greatest density in subcortical grey matter. Only a small percentage (average, 6.5 per cent) contained CMV inclusion-bearing cells. Isolated inclusion bearing cells unaccompanied by microglial nodules or inflammatory infiltrates were seen in half the patients. CMV inclusions were identified in capillary endothelia, astrocytes, and neurons. Focal CMV necrosis, ventriculo-encephalitis, and radiculo-myelitis were less frequent. The presence of CMV inclusions in capillary endothelia suggests a vascular portal of entry for the virus into the central nervous system. The diffuse ependymal and/or subpial distribution of CMV in several patients suggests additional dissemination via the cerebrospinal fluid. Isolated inclusion-bearing cells may reflect the relative nonpermissiveness of surrounding central nervous system parenchyma for CMV infection. PMID- 3028932 TI - Assignment of the prealbumin (PALB) gene (familial amyloidotic polyneuropathy) to human chromosome region 18q11.2-q12.1. AB - The assignment of the human prealbumin (PALB) gene to chromosome region 18q11 q12.1 has been achieved using a human genomic probe in the study of human-mouse somatic cell hybrids and by in situ hybridization. Because familial amyloidotic polyneuropathy was reported previously to be due to a mutation in prealbumin, it can be inferred that the gene for this disorder also maps to 18q11.2-q12.1. PMID- 3028934 TI - Adenosine deaminase, adenylate kinase and acid phosphatase polymorphism in a French-Canadian population. AB - Adenosine deaminase (ADA), adenylate kinase (AK1), and acid phosphatase (ACP1) red blood cell enzymes were studied for allelic variation in a French-Canadian population from Quebec City, Canada. Allele frequencies in 887 unrelated individuals were for ACP1, ACP1*A: 0.305; ACP1*B: 0.635 and ACP1*C: 0.060, for ADA, ADA*1: 0.969, ADA*2: 0.031, and for AK1, AK1*1: 0.976, AK1*2: 0.024. The allele frequencies for each enzyme were identical to those previously reported in other Caucasian populations. PMID- 3028935 TI - Infection with human immunodeficiency virus (HIV) and cytomegalovirus in a London health district 1980-4. AB - By testing serum samples taken between 1980 and 1984 from men attending a department of sexually transmitted diseases, it was shown that antibodies to human immunodeficiency virus (HIV) first appeared in 1981. Homosexual men were significantly more likely to have antibodies to HIV and to cytomegalovirus (CMV) than were heterosexual men attending the same clinic. This shows that homosexuals are exposed to both HIV, the cause of the acquired immune deficiency syndrome (AIDS), and to CMV, which can reactivate to cause life threatening disease once immunosuppression has developed. All homosexuals, not just those with antibodies against HIV, had raised levels of CMV antibodies. This suggests that they experience frequent antigenic stimulation after reinfections with CMV or reactivation of endogenous virus. PMID- 3028933 TI - Regional localization of DNA probes on the short arm of chromosome 11 using aniridia-Wilms' tumor-associated deletions. AB - We are interested in the precise localization of various DNA probes on the short arm of chromosome 11 for our research on the aniridia-Wilms' tumor association (AWTA), assigned to region 11p13 (Knudson and Strong 1972; Riccardi et al. 1978). For this purpose we have screened lymphocyte DNA and material derived from somatic cell hybrids from individuals with constitutional 11p deletions with a range of available probes: D11S12; calcitonin/CGRP (CALC1/CALC2); insulin (INS); Harvey ras 1 (HRAS 1); beta-globin gene cluster (HBBC); human insulin-like growth factor 2 (IGF-2); parathyroid hormone (PTH); human pepsinogen A (PGA). Using this material, it has been possible to map all probes used, except insulin, outside the region 11p111-p15.1, resulting in an SRO (same regional overlap) of 11p15.1 p15.5 for most probes. We found an SRO for PGA of 11p111-q12 and an SRO for CALC2 of 11p15.1-p15.5 or 11p111-q12. We have localised the insulin gene to band 11p15.1. PMID- 3028937 TI - Conformational alteration in foot-and-mouth disease virus virion capsid structure after complexing with monospecific antibody. AB - A mechanism of neutralization of virus infectivity by antibody is described and related to the immune defences in vivo. The interaction of a particular monoclonal antibody with homologous foot-and-mouth disease virus alters the conformation of the virions to permit penetration of staining reagents. A consequence of this structural alteration is that the RNA genome becomes susceptible to dissociation from the capsid proteins. This mechanism of virus neutralization is irreversible and therefore provides an effective in vivo defence measure against virus attack, complementing the enhanced phagocytosis effected through opsonization of virions. With viruses that can replicate in phagocytes, such a mechanism of virus neutralization could provide a major 'specific' immunological defence against virus invasion. PMID- 3028936 TI - Detection of human papillomavirus DNA in anogenital condylomata in men using in situ DNA hybridisation applied to paraffin sections. AB - An in situ DNA hybridisation method was used to detect human papillomavirus (HPV) DNA (HPV types 6, 11, 16, and 18), and an immunoperoxidase (IP-PAP) method to detect HPV structural protein expression in paraffin sections of biopsy specimens from 133 men treated for penile (in 114 cases) and anal (in 19 cases) warts. The anatomical distribution on the penis of classic condyloma acuminatum and of papular and flat condylomata was practically identical. The gross appearance of the warts did not correlate with their morphology on light microscopy. The detection rate of dysplasia was very different in the three types of lesions (25% in flat, 50% in acuminatum, and 75% in papular warts). Of 133 lesions, 59 (44.4%) contained HPV antigens, their expression being inversely related to the grade of dysplasia; only 17% of HPV 16 lesions had detectable HPV antigen compared with 50% to 67% in lesions of the other three HPV types. HPV 16 and HPV 18 DNA were most commonly (11%) detectable in Bowenoid lesions; however, most of the HPV 16 and 18 positive cases were found among the flat and acuminatum type of lesions. Though the overall detection rate of HPV DNA (76%) did not correlate with the grade of dysplasia, a clear cut association of HPV 16 and HPV 18 with dysplastic lesions was found, none of the HPV 16 and 25% of the HPV 18 positive cases being devoid of concomitant dysplasia. The corresponding figures for HPV 6 and HPV 11 were 59.2% and 68.8%, respectively. The implications of these findings are discussed in terms of epidemiologically established connections between penile and cervical cancer, with special emphasis of the high risk HPV types 16 and 18. The applicability of in situ DNA hybridisation as a powerful tool in the analysis of specific HPV DNA sequences in routinely processed biopsy specimens from these lesions is emphasised. PMID- 3028938 TI - Requirements for growth of Epstein-Barr virus-transformed cells at low cell densities. AB - We investigated the requirements for growth of Epstein-Barr virus-transformed cells at low cell densities in a homotypic system: only Epstein-Barr virus transformed cells or their products were used to supply feeder activity. The cloning efficiency was increased markedly under conditions favouring close cell to-cell contact: culture in round-bottomed rather than in flat-bottomed wells or the addition of irradiated Epstein-Barr virus-transformed cells. Further enhancement was obtained by the addition of the tumour promoter phorbol-myristate acetate. Part of this effect was also induced by supernatants of Epstein-Barr virus-transformed cells, in particular when the latter had been stimulated with phorbol ester. Thus, under limiting dilution conditions, Epstein-Barr virus transformed cells were found to be dependent upon autologous cell-mediated and soluble proliferation-stimulating signals. In such a homotypic feeder system, the cloning efficiency could be enhanced up to eight-fold; a 100% cloning efficiency could not be achieved. Evidence is presented that the residual deficit in cloning efficiency is inherent to these cell lines. PMID- 3028940 TI - Specific inhibition of macrophage antibody-dependent endocytosis by p chloromercuribenzenesulphonic acid: identification of sensitive membrane proteins. AB - Murine macrophage Fc receptor function has been studied with the membrane impermeable sulphydryl-blocking reagent p-chloromercuribenzenesulphonic acid (PCMBSA). Antibody-dependent endocytosis of fluorescein-labelled immune complexes was studied with video intensification microscopy. PCMBSA was found to inhibit the endocytosis of immune complexes at 10 nM. Control experiments indicate that the inhibition is due to an effect upon the cell, not the immune complex. Furthermore, specificity is suggested by the fact that complement-mediated and latex bead phagocytosis were not affected. 203Hg-PCMBSA labelled three macrophage proteins of molecular weight (MW) 25,000, 35,000 and 50,000. A 25,000 MW PCMBSA binding protein has been found that is specifically immunoprecipitated by an anti Fc receptor antibody. These studies suggest that perturbation of a cell surface sulphydryl group(s) of one of the three major PCMBSA binding membrane proteins, possibly an Fc receptor-associated protein, blocks a molecular signal required for antibody-dependent endocytosis. PMID- 3028939 TI - Frequency of B-lymphocyte transformation by Epstein-Barr virus decreases with entry into the cell cycle. AB - The relationship between in vitro B-cell activation and transformation by Epstein Barr virus (EBV) was studied. B cells were fractionated using discontinuous Percoll gradients to purify cells with resting morphology. Activation of resting cells for 24 hr with anti-Ig (mu chain specific) or Staphylococcus aureus Cowan I (SAC) resulted in transition of susceptible cells into the G1 phase of the cell cycle as shown by an increase in cell size, an increase in uridine incorporation and an increase in sensitivity to B-cell growth factor (BCGF). Entry into S phase was achieved by extending the period of activation to 48-96 hr with high concentrations of SAC or anti-mu or using BCGF. SAC-activated cells entered S phase on Day 2 and anti-mu treated cells on Day 3. Control (G0) cells and cell activated for varying lengths of time (G0/G1, G1/S) were exposed to EBV and plated in a limiting dilution assay to determine the frequency of EBV transformable cells. Control cells and cells activated for 24 hr had a transformation frequency of 1-2%. With continued activation with SAC or anti-mu, however, transformation frequency decreased at a rate paralleling the entry of the population into S phase. Treating cells with low concentrations of anti-mu or SAC in combination with BCGF decreased the transformation frequency to levels lower than anti-mu or SAC alone, further suggesting that entry into S phase is accompanied by a reduction in transformability. These results indicate that resting B cells are highly susceptible to transformation, and that with in vitro activation into the cell cycle B cells become resistant to EBV transformation. PMID- 3028942 TI - Mechanism of neuromuscular blocking activity of 1,3-bis-(3-t-butylamino-1 propoxy)benzene dimethiodide (compound 79-297). PMID- 3028941 TI - Specific stimulation of lymphocytes from patients with AIDS by herpes simplex virus antigens. AB - Peripheral blood lymphocytes of patients with AIDS are generally anergic to stimulation by lectins or by alloantigens. We have succeeded in demonstrating that the lymphocytes of six AIDS patients, with histories of herpes simplex virus (HSV)-induced genital, anal and/or oral lesions, retained functional specificity with regard to HSV antigens. This was accomplished by treating patient lymphocytes with OKT8 monoclonal antibodies, in the presence of complement, to yield a cell population that was partially responsive to phytohaemagglutinin (PHA). Enhanced proliferation was obtained in the presence of exogenous interleukin-2, which was also employed to foster the growth of these cells over several generations. Lymphocytes derived from these AIDS patients, and maintained in vitro as described, were specifically stimulated in proliferation assays by a HSV antigen preparation in six of the ten cases studied. PMID- 3028943 TI - Influence of follicle stimulating hormone on phosphomonoesterases and adenosine triphosphatases in the ovary of immature bonnet monkeys (Macaca radiata, Geoffroy). PMID- 3028944 TI - Prolonged survival (more than 3 years) in a case of small cell carcinoma of the lung. PMID- 3028946 TI - A survey of rotavirus antibody in different age groups. PMID- 3028945 TI - Genetic organization of the ovine MHC class II region. AB - To study the class II genes of the major histocompatibility region of the sheep genome, human HLA class II genes corresponding to the known subregions in man (DR, DQ, DP, DO, and DZ) were used for Southern hybridization analysis of sheep DNA and to probe a sheep genomic library. Hybridizing bands were noted for all probes except DP alpha. DQ alpha and beta and DR beta appear to be present as multicopy genes, while DR alpha-, DZ alpha-, and DO beta- like genes appear to be single copy. All bands detected with the DP beta probe were also detectable with other beta chain probes. From eight lambda-bacteriophage clones of a sheep genomic library nine distinct class II genes were identified. These genes were characterized by differential hybridization analysis and restriction mapping. Two genes were DR beta-like, three DQ alpha-like and four DQ beta-like. The extensive cross-hybridization observed with beta chain probes was not seen with alpha chain probes. The results of this study suggest that the major histocompatibility complex class II region of the sheep has a similar genetic organization to that of man, with the provisional exception of the DP subregion. PMID- 3028947 TI - Comparison of reverse passive haemagglutination assay & solid phase agglutination of coated erythrocytes with ELISA for rotavirus antigen detection. PMID- 3028948 TI - Liver tumours of infancy and childhood in south Orissa. PMID- 3028949 TI - Pleomorphic adenoma of skin (chondroid syringoma). PMID- 3028950 TI - Malanotic neuroectodermal tumor of infancy. A case report. PMID- 3028951 TI - Primary liver cell carcinoma and hepatitis B antigen (a clinicopathological study). PMID- 3028952 TI - Japanese encephalitis: containing the growing threat. PMID- 3028953 TI - Comparison of a new renin inhibitor and enalaprilat in renal hypertensive dogs. AB - The hypotensive efficacy of a potent new renin inhibitor (N alpha-isovaleryl-L histidyl-L-prolyl-L-phenylalanyl-L-histidyl-ACHPA+ ++-L- phenylalanyl amide) containing (3S,4S)-4-amino-5-cyclohexyl-3-hydroxy pentanoic acid (ACHPA) was compared with the converting enzyme inhibitor (enalaprilat) (MK-422) in conscious one-kidney dogs before and after tightening a renal artery clamp. Dose-response curves to 0.003 to 0.1 mg/kg/min i.v. infusions of the ACHPA-containing renin inhibitory peptide or enalaprilat (0.003-0.1 mg/kg i.v. bolus) were obtained in one-kidney dogs before and 3 days and 14 days after renal artery constriction. The ACHPA-containing renin inhibitory peptide and enalaprilat maximally decreased blood pressure by 10 +/- 2 and 9 +/- 2 mm Hg before constriction and by 12 +/- 2 and 12 +/- 4 mm Hg in dogs treated 14 days after renal artery constriction, respectively. Glomerular filtration rate and effective renal plasma flow were unaltered or slightly improved. In sharp contrast, both compounds elicited significant, dose-related decreases in blood pressure (-26 +/- 4 and -20 +/- 4 mm Hg, respectively), glomerular filtration rate (-21 +/- 3 and -23 +/- 3 ml/min), and renal plasma flow (-45 +/- 14 and -48 +/- 13 ml/min) in dogs examined 3 days after renal artery constriction. These data demonstrate that ACHPA-containing renin inhibitory peptide and enalaprilat are equally effective antihypertensive agents in dogs with renin-dependent renovascular hypertension and lend credence to the contention that the renin-angiotensin system supports renal function in hypertensive states in which renin levels are elevated. PMID- 3028954 TI - A factor that initiates myocardial hypertrophy in hypertension. AB - A lack of correlation between blood pressure and myocardial hypertrophy was established in spontaneously hypertensive rats, suggesting that factors other than blood pressure control might be responsible for the modulation of myocardial hypertrophy. An in vitro system that is independent of blood pressure and hemodynamic effects was developed by use of isolated myocytes to study myocardial protein synthesis. The validity of this system was determined by means of morphology, by receptor integrity, and by studying the incorporation of tritiated leucine into myocyte protein (dpm/mg/hr). Addition of a supernatant of spontaneously hypertensive rat myocardial homogenate (centrifuged at 1500 g) to the myocyte system resulted in a significant increase in tritiated leucine incorporation into myocyte protein when compared with the addition of homogenates from normal controls. The protein from the homogenate was partially purified by high performance liquid chromatography. The resultant purified protein also stimulated protein synthesis by 70%. Furthermore, a significant increase in the specific activity of the transfer RNA and the rate of protein synthesis was observed after addition of homogenate from hypertrophied heart (4.02 +/- 0.3 vs 7.0 +/- 0.2 pmol leucine/microgram protein/hr; p less than 0.05). These data demonstrate the existence of a soluble factor in the hypertrophied myocardium that stimulated protein synthesis. This factor may play a key role in modulation of myocardial structure during development or regression of myocardial hypertrophy in hypertension. PMID- 3028955 TI - Increased vasodilator responses to acetylcholine in psychosocial hypertensive mice. AB - Responsiveness to endothelium-dependent (acetylcholine and A23187) and endothelium-independent (nitroprusside and 8-bromo cyclic guanosine 3',5' monophosphate [cGMP]) vasodilators was examined in two vascular preparations from hypertensive and normotensive mice. CBA Agouti mice were made hypertensive by exposure to social stress in a complex population cage. After 2 months, the hindquarter vascular bed was pump-perfused at a constant flow with plasma substitute to evaluate changes in perfusion pressure, and helical strips of aorta were suspended in muscle baths for measurement of isometric force generation. Tissues were treated with methoxamine to induce contractile tone. Threshold dilator responses to acetylcholine were elicited at a significantly lower dose in the hindquarters of hypertensive mice than in those from normotensive mice, indicating increased vasodilator sensitivity. In contrast, vasodilator responsiveness to nitroprusside in hindquarters of hypertensive mice did not differ from that in hindquarters of normotensive mice. Aortas from hypertensive mice were more sensitive (lower ED50) to the relaxant effects of acetylcholine and A23187 than those from normotensive mice. The relaxant effects of nitroprusside and 8-bromo cGMP on aortas from hypertensive mice were not significantly different from those in normotensive aortas. Aortic strips that had been rubbed on the lumen surface with a wooden stick did not relax to acetylcholine or A23187. In aortas that were not initially contracted with methoxamine, acetylcholine and A23187 caused small contractions from baseline. The magnitude of these contractile responses were potentiated after removal of the endothelium, and the potentiation was greater in aortas from hypertensive mice. These results demonstrate an increased responsiveness to endothelium dependent vasodilators in this psychosocial model of hypertension. PMID- 3028956 TI - Lymphocyte membrane sodium-proton exchange in spontaneously hypertensive rats. AB - The sodium-proton exchange activity was determined in lymphocytes of spontaneously hypertensive rats (SHR), normotensive Wistar-Kyoto rats (WKY), and domestic Wistar rats. Uptake of sodium was determined by measuring the osmotic swelling of lymphocytes after activation of the exchanger by suspension of the cells in sodium propionate and consequent intracellular acidification by the permeant weak acid. Fractional swelling (mean +/- SEM) in 16 SHR and 16 WKY was 0.44 +/- 0.03 and 0.35 +/- 0.02, respectively (p less than 0.01). The swelling was partially inhibitable by amiloride and, at 10(-4) M concentration, the amiloride-sensitive swelling was 0.21 +/- 0.02 in SHR and 0.11 +/- 0.01 in WKY (p = 0.001). Progressive extracellular ion substitutions of chloride for propionate or of potassium for sodium showed that the exchange activity was related linearly to cellular acidification; however, the dependence on extracellular sodium displayed saturation characteristics, with the same apparent Km for cells from SHR and WKY and a Vmax of 0.54 +/- 0.03 for SHR and 0.39 +/- 0.02 for WKY (p less than 0.002). External lithium could replace sodium on the exchanger but abolished the differences between strains. Results in the domestic Wistar rats were similar to those of WKY. These results suggest that lymphocytes of the SHR have a greater capacity for sodium uptake through the sodium-proton exchanger, as compared with normotensive strains. If shared by other cells, such an increased capacity could have a pathophysiological role in genetic hypertension. In particular, its presence in proximal renal tubular cells would support the hypothesis of a primary role for the kidney in the pathogenesis of genetic hypertension. PMID- 3028957 TI - Role of angiotensin II in the hormonal, renal, and electrolyte response to sodium restriction. AB - Adrenal responses to angiotensin II (ANG II) are enhanced with restriction of sodium intake. To determine whether increased circulating ANG II levels are responsible for the enhanced responsiveness, the adrenal and blood pressure responses to ANG II in human subjects were assessed four times: in balance on a high and a low salt diet and before and after the administration of a converting enzyme inhibitor (enalapril). Before enalapril administration, sodium restriction significantly increased (p less than 0.02) plasma renin activity, ANG II, and aldosterone levels; the aldosterone response to ANG II was enhanced twofold (p less than 0.01); and the blood pressure response to ANG II infusion was reduced significantly (p less than 0.05). Despite a fixed and low plasma ANG II concentration when enalapril was employed, the adrenal response to ANG II on the low salt diet was enhanced to the same degree as that observed before administration of the converting enzyme inhibitor. Conversely, enalapril substantially altered the blood pressure response to ANG II with sodium restriction, completely preventing the reduction in responsiveness. If the subjects were first given enalapril and then sodium intake was restricted, ANG II levels did not change significantly but renal excretion of both sodium and potassium was substantially modified. The rate at which renal excretion of sodium fell to match intake was retarded strikingly (p less than 0.001); conversely, renal retention of potassium increased significantly (p less than 0.03) as low salt balance was attained. Possibly because of the potassium retention, aldosterone levels rose, but significantly less than when enalapril was absent. PMID- 3028959 TI - Effect of ACTH on glucose uptake by hepatic cells and its effect on hepatic enzymes in presence of insulin and acetylcholine. AB - An intrinsic hypoglycaemic activity has been attributed to ACTH. This action of ACTH is essentially mediated by increased insulin secretion. In the present study, the direct action of ACTH on glucose uptake by pigeon hepatocytes has been studied by in vitro technique. The results have shown that ACTH has a direct influence on glucose uptake and this action is not additive in presence of insulin. Glucose uptake in presence ACTH with Acetylcholine was not any more than what was obtained with ACTH alone. These observations have been taken to indicate a non-existence of synergistic action of ACTH with insulin or acetylcholine in promoting glucose uptake by avian liver cells. PMID- 3028958 TI - Cataracts and hypertension in salt-sensitive rats. A possible ion transport defect. AB - In previous unrelated studies, we observed a 35 to 50% incidence of cataract formation in several groups of Dahl salt-sensitive hypertensive rats (DS) over a 4-year period. In the present study we evaluated longitudinal changes in blood pressure in DS in which cataracts eventually developed and those in which cataracts did not develop when all animals were maintained on a high sodium diet. Lenses were evaluated by slit-lamp microscopy to determine if cataractous lesions were similar among rats, to classify lesion types, and to define the age at which cataracts were detectable in DS. The possible participation of several cataractogenic risk factors as major influences on cataract formation also was evaluated. Finally, aqueous humor concentrations and lenticular content of sodium and potassium were determined to evaluate the possibility that a defect in ion transport at the lens epithelium and ciliary body might play a role in cataractogenesis in DS, since ion transport defects have been shown to lead to lens opacification in other models of genetic and experimental cataracts. Parallel studies were performed in Dahl salt-resistant control rats (DR). A high incidence of cataract formation was found in DS. Although systolic blood pressure was not consistently greater in adult DS with cataracts compared with values in age-matched DS without cataracts, the initial pressor response to a high salt diet was greatest in weanling DS in which cataractous lesions later developed. Slit-lamp analysis revealed that cataracts in this genetic model were cortical, with one mixed cortical, nuclear lesion. Posterior subcapsular lesions were not observed, suggesting that lesions were not steroid-induced.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3028960 TI - Peripheral action of angiotensin II. AB - The effects of angiotensin II were studied on isolated atrial preparations of nonreserpinised and reserpinised rabbits, before and after treating the preparations by propranolol. Peripheral action of angiotensin was cardioaccelerator via direct stimulation of beta-receptors of the atria in isolated atrial preparations. PMID- 3028961 TI - In vitro effect of synthetic pyocyanine on neutrophil superoxide production. AB - Pyocyanine, a low-molecular-weight phenazine pigment produced by Pseudomonas aeruginosa, has previously been shown to strongly inhibit human lymphocyte blastogenesis. We now report that synthetic pyocyanine can also affect the generation of superoxide by human peripheral blood polymorphonuclear leukocytes (PMNs) in a dose-dependent manner. Superoxide production by PMNs stimulated with phorbol myristate acetate (PMA) was measured in the presence and absence of pyocyanine, phenazine, and trifluoperazine, a phenothiazine of similar chemical structure to the phenazine pigments. Pyocyanine at 50 microM inhibited superoxide production to 28.9 +/- 2.8% of PMA control values, whereas at the lower concentration of 1 microM, the production of superoxide was significantly enhanced (203 +/- 31.7% of PMA control values). Phenazine, the tricyclic parent compound of pyocyanine, had only a minor effect. Trifluoperazine had a marked inhibitory effect on superoxide generation at concentrations above 1 microM. None of the compounds induced superoxide generation in the absence of PMA. Pyocyanine at all concentrations, unlike phenothiazines, had very little effect on the release of neutrophil granule enzymes. The effect of P. aeruginosa phenazine pigments on polymorphonuclear phagocytes is of significance, since inhibition of host PMN function at sites of infection could result in ineffective bacterial killing, whereas enhanced PMN function could lead to greater tissue damage. These two possibilities are not mutually exclusive and may coexist depending on local pyocyanine concentrations. PMID- 3028963 TI - An outer membrane protein (porin) as an eliciting antigen for delayed-type hypersensitivity in murine salmonellosis. AB - The porin, an outer membrane protein of Salmonella typhimurium, was found to be a suitable antigen for eliciting delayed-type hypersensitivity in mouse salmonellosis. Histological examination of the reaction site revealed that the porin was superior to other antigenic preparations in eliciting a typical delayed type hypersensitivity reaction consisting of mononuclear cell infiltration without polymorphonuclear cell contamination. This study indicates the importance of using a suitable protein antigen from S. typhi for human application. PMID- 3028962 TI - Virulence of iron transport mutants of Shigella flexneri and utilization of host iron compounds. AB - Mutants of Shigella flexneri defective in aerobactin-mediated iron transport were assayed for virulence in several model systems. A Tn5 insertion mutant was invasive in HeLa cells, lethal in the chicken embryo, and produced keratoconjunctivitis in the guinea pig, indicating little or no loss of ability to invade and multiply intracellularly. Although the mutant failed to grow in low iron medium in vitro, growth equivalent to that of the wild type was observed in HeLa cell lysates. Thus, there appears to be sufficient available iron inside the HeLa cell to allow growth in the absence of siderophore synthesis. Possible host iron sources were tested, and both the mutant and wild type utilized hemin or hematin as a sole source of iron. Only the wild-type, aerobactin-producing strain could remove iron from transferrin or lactoferrin. Two deletion mutants were also assayed for virulence and were found to be avirulent for the chicken embryo. These deletions encompass flanking sequences as well as the aerobactin genes; therefore, adjacent genes may be required for virulence. PMID- 3028964 TI - Bacteriological data on a prospective multicenter study of the effect of two different regimens for selective decontamination in patients with acute leukaemia. AB - In this paper we described the results of bacteriological monitoring of oropharynx and stool samples from granulocytopenic patients with leukaemia who received oral infection prophylaxis with two different regimens for selective decontamination of the digestive tract. Patients were prospectively randomized either into a group receiving non-absorbable antimicrobial drugs for selective decontamination (polymyxin and neomycin: group A) or into a group receiving polymyxin and co-trimoxazole (group B). The oropharynx was, or became, free of gram-negative bacilli within one week of treatment in 94% and 90%, respectively, of the patients in group A and group B. The stool samples were, or became, negative after the same treatment interval in 91% and 80%, respectively, of the two patient groups. Antibiotic therapy during selective decontamination treatment significantly increased the incidence of positive cultures from the oropharynx and stools. The sensitivity of the gram-negative bacilli isolated during selective decontamination treatment to the drugs administered did not influence the average response to treatment. Both resistant and sensitive gram-negative bacteria appeared to disappear from the patients' samples, mostly within a week, without the need to adjust the selective decontamination treatment. Yeasts behaved in almost the same way as gram-negative bacilli. All patients received oral amphotericin B; some patients occasionally yielded oropharyngeal or faecal cultures which were positive for yeasts. PMID- 3028965 TI - International experiences with ofloxacin, a new quinolone. AB - In vitro and in vivo antibacterial activities of ofloxacin, a new quinolone, are presented. Ofloxacin was found to be characterized by potent bactericidal activity due to its strong inhibition of bacterial DNA gyrase. The high antibacterial activity of ofloxacin in vivo was discussed with respect to its physiochemical properties, i.e., hydrophobicity and bioavailability including high serum concentrations and easy entry into bacterial cells. PMID- 3028966 TI - Antibacterial activity of ofloxacin and its mode of action. AB - The antibacterial activity of ofloxacin against Enterobacteriaceae, Pseudomonas aeruginosa, Haemophilus influenzae, Branhamella catarrhalis, and Neisseria gonorrhoeae was comparable to norfloxacin and enoxacin, and far exceeded the activity of pipemidic acid and nalidixic acid. The activity of ofloxacin was two to eight times less than that of ciprofloxacin. Ofloxacin was more active against Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus pneumoniae, Acinetobacter spp., Legionella spp., and Bacteroides fragilis, than norfloxacin, enoxacin, pipemidic acid and nalidixic acid, and the activity of ofloxacin was comparable to that of ciprofloxacin. Ofloxacin was two to seven times more effective than norfloxacin in systemic infections in mice with S. aureus, Escherichia coli, Serratia marcescens and P. aeruginosa. Ofloxacin strongly inhibited DNA supercoiling activity of DNA gyrase purified from E. coli KL-16. There is a parallel relationship between antibacterial activity of ofloxacin and its inhibitory action against DNA gyrases from ofloxacin-susceptible and ofloxacin-resistant clinical isolates of E. coli. These results indicate that the high bactericidal action of ofloxacin and the related new quinolone agents can be explained by their potent inhibitory activities against DNA gyrase in bacterial cells. PMID- 3028967 TI - Delta agent and the etiology of hepatocellular carcinoma. AB - Sera from 87 patients with hepatitis B surface antigen (HBsAg)-positive hepatocellular carcinoma (HCC) and from 29 HBsAg-positive hospital controls were tested for delta (delta) antigen with an immunoenzymatic procedure and for anti delta antibody, hepatitis Be antigen (HBeAg) and antibody to HBeAg (anti-HBe) by radioimmunoassay. All the sera, from both the HCC cases and the control patients, were negative for delta-antigen. Among the HCC cases 9 were positive for serum anti-delta (10%) whereas among the controls none was positive for this antibody (p = 0.067); the anti-delta-positive cases were found only among HCC patients negative for HBeAg. The lower prevalence of anti-delta among HBsAg-positive HCC patients, as compared to the corresponding prevalence among HBsAg-positive patients with chronic active hepatitis or cirrhosis (reported in the literature) indicates that the pathogenesis of HCC is frequently independent of the pathogenesis of the other HBsAg-positive common chronic liver diseases. By contrast, the higher prevalence of anti-delta among HBsAg-positive HCC cases than among HBsAg-positive controls may reflect the longer average duration of the carrier state in HCC patients (until integration is accomplished and the induction period completed). Serum HBeAg was higher among HBsAg-positive HCC patients with cirrhosis (23%) than among HBsAg-positive HCC patients without cirrhosis (6%) or HBsAg-positive controls (3%); thus, the conflicting results in the literature concerning the association of the HBeAg/anti-HBe system with HCC may be accounted for, in part, by the variable association of HCC with an underlying cirrhosis. PMID- 3028969 TI - Qualitative and quantitative evaluation of chrysotile and crocidolite fibers with IR-spectroscopy: application to asbestos-cement products. AB - Infrared (IR) spectrophotometry allows simple and quick qualitative and quantitative evaluations of different kinds of asbestos, as well as of other inorganic particles. In particular, chrysotile and crocidolite have characteristic IR spectra and optical density measures of 2,710 nm band for chrysotile, of 12,820 nm band for crocidolite permit quantitative evaluation of each fiber alone or in mixture. IR spectra also give informations about changes of fiber structure and of chemical composition due, for example, to thermal treatment or acid leaching. The analytical method we developed can detect levels as low as 0.1 mg of fiber in a 300 mg disk of KBr using a low cost IR spectrophotometer. The use of a Fourier Transform IR spectrophotometer (FTIR) improves dramatically the sensitivity and selectivity. Computer assisted analysis of spectra offers the possibility to reduce matrix interferences and to compare different spectra. Examples of IR technique applied to asbestos-cement products and insulating materials are presented. PMID- 3028968 TI - Effect of primrose oil on serum lipids and blood pressure in hyperlipidemic subjects. AB - Primrose oil (about 72% of fatty acids 18:2 omega 6 and 9% of 18:3 omega 6) was supplemented in addition to the used diet of 20 hyperlipidemic patients for 3 months in an open study. The dose of primrose oil was 2.4 ml, 4.8 ml and 7.2 ml during the first, second and third month, respectively. Primrose oil increased the proportion of gamma-linolenic acid in serum cholesteryl esters (1.81 vs. 2.13%, p less than 0.05), phospholipids (0.65 vs. 0.82%, p less than 0.01), triglycerides (0.70 vs. 0.99%, p less than 0.01) and free fatty acids (0.80 vs. 0.91%, NS). It did not, however, change serum cholesterol, HDL cholesterol or triglyceride mean values. Blood pressure mean values remained unchanged during the follow-up as well. PMID- 3028970 TI - Characteristics of women under 20 with cervical intraepithelial neoplasia. AB - The relationship between risk of cervical neoplasia and various sociodemographic, reproductive and sexual characteristics was studied in 126 women (cases) with an abnormal Papanicolaou test finding and 1914 controls seen at the same clinic in which cases were detected but showing no evidence of cervical neoplasia. All the subjects of this study were under the age of 20. Number of sexual partners and clinical occurrence of genital warts emerged as the most important determinants of the risk of cervical intraepithelial neoplasia (CIN) in these very young women (Relative Risk (RR) for three or more sexual partners = 2.45, 95% Confidence Interval (CI) 1.63-3.70, RR for occurrence of genital warts, adjusted for number of sexual partners = 9.15, 95% CI: 5.06-16.26). Also having grown up in a 'problem' family seemed to increase the probability of developing CIN (RR adjusted for number of partners = 1.64, 95% CI 1.06-2.52). For all other investigated characteristics, cases and controls were remarkably similar. These include socioeconomic status, parity, smoking habits, use of various contraceptive methods and also indicators of sexual activity such as age of first having sexual intercourse, and duration and frequency of intercourse. Although this group of women under 20 only allows the study of less severe precursors of cervical cancer, it helps to highlight the earliest effect of purported risk factors for cervical neoplasia, chiefly sexual habits, at a time very close to their being established. PMID- 3028971 TI - Infections due to the human herpesviruses in southern India: a seroepidemiological survey. AB - We studied the prevalence of IgG and IgM antibodies to the human herpesviruses in a hospital-based population of 181 individuals aged 0 to 25 years, who were resident in Vellore, south India or surrounding rural areas. Antibodies to the Epstein-Barr virus (EBV) viral capsid antigen were determined by indirect immunofluorescence, while antibodies to the remaining herpesviruses were determined by enzyme-linked immunosorbent assay. The age-specific prevalence rates of IgG antibodies to EBV and cytomegalovirus (CMV) rose rapidly after birth to reach a value of over 90% by the fourth year of life. High age-specific IgM prevalence rates and geometric mean titres (GMT) of IgG antibody in children 6 months to 2 years of age, and the early median age of virus infection (1.4 years for EBV and less than 1 year for CMV) indicate that primary infection with these viruses occurs early in life. In contrast, age-specific prevalence rates of IgG antibodies to varicella-zoster virus (VZV) and herpes simplex virus (HSV) rose gradually after birth to attain maximal values of only 72% (VZV) and 83% (HSV) in the 15-25 year age group, and the median ages of infection were delayed (12.25 years for VZV and 8.2 years for HSV). The age-specific IgG prevalence rates of VZV and HSV, and of EBV and HSV showed statistically significant positive correlations, suggesting that common epidemiological factors may underlie the pattern of infections due to these groups of viruses.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3028972 TI - Semisynthesis of the native sequence of horse heart cytochrome c-(66-104) nonatriacontapeptide. AB - The syntheses of some derivatives of horse cytochrome c-(66-79)-tetradecapeptide are presented. The syntheses are so designed that analogues of this phylogenetically well preserved sequence can be obtained also. The compounds were intended as synthons for the semisynthesis of 65-homoserine-cytochrome c, which we described earlier. A requisite for this project was the C-terminal tetracosapeptide fragment of the protein, accessible through degradation of cytochrome c with cyanogen bromide. The five epsilon amino groups in this compound are reversibly protected with the 2-(methylsulfonyl)ethyloxycarbonyl function, which is resistant to acid and causes little impairment of solubility. The condensation of the fragments leading to the native sequence of horse cytochrome c-(66-104)-nonatriacontapeptide is presented also. The syntheses were performed using the solution strategy. Some unexpected ring closing reactions involving tyrosine and tert.-butyl prolylasparaginylcarbazate, are described. PMID- 3028973 TI - Side chain to side chain cyclization of an enkephalin analog results in loss of opioid receptor selectivity. AB - The cyclic enkephalin analog H-Tyr-D-Lys-Gly-Phe-Glu-NH2 (I) and the structurally related open chain analogs H-Tyr-D-Nle-Gly-Phe-Gln-NH2 (II) and H-Tyr-D-Lys(For) Gly-Phe-Abu-NH2 (III) were tested in mu and delta opioid receptor-representative binding assays and bioassays. Whereas both linear analogs showed a pronounced preference for mu receptors over delta receptors, the conformationally restricted cyclic peptide I was found to be unselective. This finding represents the first reported example of a peptide cyclization resulting in a loss of receptor selectivity. From this and earlier studies, it was concluded that the receptor selectivity of cyclized peptide analogs relative to that of their linear correlates may depend on the size and relative rigidity of their ring structures. PMID- 3028974 TI - Oxygen-18 isotope labeling and its effect on carbon-13 chemical shifts of the peptide bond. AB - The effects of 18O isotopes on 13C NMR chemical shifts of peptide carbonyl carbons were found to be 0.028 ppm for glyclyglycine and 0.029 ppm for glycylproline at 50.31 MHz. PMID- 3028975 TI - Beta-lactamase inhibitors: another approach to overcoming antimicrobial resistance. AB - Beta-lactamase inhibitors in combination with a beta-lactam antimicrobial agent provide another approach to the treatment of infections caused by many bacteria resistant to commonly used agents. Currently two fixed combinations are commercially available. One is the oral preparation of amoxicillin and potassium clavulanate (Augmentin). The other is for intravenous therapy and combines ticarcillin and potassium clavulanate (Timentin). In vitro studies confirm that the combination of a beta-lactamase inhibitor and a beta-lactam can increase the activity of the beta-lactam to such important pathogens as S aureus, Klebsiella, H influenzae, Bacteroides spp and other bacteria which produce plasmid-mediated beta-lactamase. Clinical efficacy studies have demonstrated potential usefulness of these combinations particularly for therapy of suspected or proven infections caused by mixed aerobic and anaerobic bacteria. The combination of amoxicillin or ticarcillin with clavulanic acid can be used to improve cost-effectiveness of antimicrobial therapy if they replace regimens that use multiple drugs or drugs with a relatively high incidence of adverse reactions. PMID- 3028976 TI - [Detection of herpes simplex virus antigen with an enzyme immunoassay and direct immunofluorescence]. AB - A total of 464 clinical specimens from patients with extragenital and genital lesions and from the cervix of asymptomatic women were examined for the presence of Herpes-simplex-virus antigen by using a commercially available enzyme immunoassay Kit (ELISA, Dakopatts) and a direct immunofluorescence technique with monoclonal antibodies (Syva-Merck). The detection rate was compared to the isolation rate of the tissue-culture technique. In extragenital lesions the sensitivity of the ELISA technique and immunofluorescence was determined to be 78.9% and 80% and in genital lesions 46.2%. In cervical clinical specimens, the sensitivity of both techniques was determined to be 5.7% and 15.4%. An HSV infection, negative in tissue culture, was diagnosed in 22 and 27 cases, respectively, using ELISA and immunofluorescence. This diagnosis was confirmed in 21 and 22 cases by the other immunological technique. For this reason, it can be assumed that in some cases direct immunological techniques can be more sensitive than the tissue culture technique. With regard to asymptomatic and manifest infections, reliable results are only obtainable in cases of manifest disease with blisters or scabs. PMID- 3028978 TI - An integrating thermoluminescent Rn daughter personal dosimeter. AB - An integrating Rn daughter dosimeter using conventional Dy-doped CaSO4 thermoluminescent (TL) material has been developed for use as a personal monitor. The dosimeter was found to have a linear response over the range 0.5 WL-h to 25 WL-h and to be insensitive to the state of equilibrium of the Rn daughters. The use of the dosimeter to monitor both 222Rn and 220Rn daughters is described. The limiting sensitivity of the dosimeter, at a sampling rate of 1 L min-1, was determined to be 0.1 WL-h for 222Rn daughters and 0.5 WL-h for 220Rn daughters. The application of the thermoluminescent dosimeter (TLD) to occupational monitoring of 222Rn and 220Rn daughters is discussed. PMID- 3028977 TI - [Stewart-Bluefarb syndrome (kaposiform arteriovenous fistula with bone changes)]. AB - In a 14-year-old female patient with arteriovenous shunts (Stewart-Bluefarb syndrome), fist-sized cutaneous swelling, venous dilatation, blue-brown pigmentation and painful subcutaneous fingertip-sized nodules were noted on the right lower thigh. In addition to an arteriovenous malformation, bone hypertrophy, bone atrophy and after bone defects were observed in the involved area on X-ray examination. Histological examination revealed increased vascularity and vascular cavernous structures with endothelial swelling and extravasation of erythrocytes, but no malignant changes. PMID- 3028980 TI - Serendipitous dosimetry--an opportunity and an opportunity lost. PMID- 3028979 TI - Cigarette smoke and lung cancer. PMID- 3028981 TI - How to differentiate neoplastic fever from infectious fever in patients with cancer: usefulness of the naproxen test. PMID- 3028983 TI - Synthesis of no-carrier-added N-([18F]fluoroalkyl)spiperone derivatives. AB - 3-N-([18F]fluoroalkyl)spiperone derivatives (2,3) can be prepared by N-alkylation of spiperone (1) with fluoroalkyl halides. The fluoroalkylating species 2 [18F]fluoroethyl bromide (7), 3-[18F]fluoropropyl bromide (8) and 4 [18F]fluorobutyl bromide (9) were prepared by [18F]fluoride ion displacement of the corresponding trifluoromethanesulfonates (triflates 4,5,6). By this method, the 2-[18F]fluoroethyl-, 3-[18F]fluoropropyl-, and 4-[18F]fluorobutyl spiperone derivatives (2a-c) can be prepared and purified rapidly and conveniently, within 40 min, in yields of 30-40% (end of synthesis, EOS). An alternative approach, suitable for the preparation of 2-[18F]fluoroalkyl (ethyl, propyl, butyl, pentyl and hexyl) spiperone derivatives (3a-d), involves iodo[18F]fluorination of terminal olefins, followed by N-alkylation of spiperone. This sequence is less convenient and proceeds in lower overall yields (less than 5%). PMID- 3028982 TI - Adenovirus pneumonia in healthy adults. PMID- 3028984 TI - Syntheses and D2 receptor affinities of derivatives of spiperone containing aliphatic halogens. AB - The development of a high affinity dopamine receptor ligand labeled with the positron emitting radionuclide, 18F (t 1/2 = 110 min), is of considerable interest for imaging and quantification of dopamine receptors in vivo. Derivatives of spiperone, a dopamine antagonist, labeled with 18F have been prepared, but the syntheses either proceed with inefficient fluoride utilization or involve several synthetic steps subsequent to 18F incorporation. To date, only the short-lived radioisotope of carbon, 11C (t 1/2 = 20.4 min), has been efficiently incorporated in the final synthetic step of 3-N-[11C]methyl spiperone. 3-N-Fluoroethyl, 3-N-chloroethyl, and 3-N-bromoethyl spiperone derivatives are prepared by alkylation of spiperone with the appropriate 2 tosyloxy ethyl halide. In addition, alpha-fluorospiperone, containing fluorine alpha to the butyrophenone carbonyl, has been prepared. The 3-N-haloethyl spiperones display high affinity for dopamine receptor in vitro. Incorporation of [18F]fluoride during the final synthetic step yields a high affinity, 18F-labeled dopamine receptor-binding ligand. PMID- 3028985 TI - The preparation of high specific activity copper-64 for medical diagnosis. AB - The specific activity of 64Cu produced by the neutron irradiation of copper is too low to be used in some in vitro tests for metabolic diagnosis. This report describes the use of the Szilard-Chalmers reaction to increase the specific activity to useful levels. PMID- 3028986 TI - External detection of beta-adrenoreceptors with 125I-hydroxybenzylpindolol in isolated perfused hearts. AB - To assess the feasibility of characterizing beta-adrenoreceptors externally with ligands labeled with gamma-emitting radionuclides, we infused 125I hydroxybenzylpindolol (125I-HYP) a potent, high affinity, beta-adrenoreceptor ligand into isolated perfused hearts and monitored uptake and release of the label externally with a gamma probe. Perfusion with specific beta-adrenergic displacing significantly increased the turnover rate constant of 125I-HYP in comparison to that obtained with non-specific displacing agents. After modifying beta-adrenoreceptor density by pretreatment of animals with thyroid hormone, an increase in 125I-HYP uptake was observed. Thus, beta-adrenoreceptor occupancy and increases in beta-adrenoreceptor density can be detected externally with an approach which may offer promise for their characterization in vivo with radiolabeled adrenergic ligands. PMID- 3028987 TI - Cytochemical localization of nicotinamide adenine dinucleotide phosphatase (NADPase) in bovine Leydig cells. AB - The ultrastructural localization of nicotinamide adenine dinucleotide phosphatase (NADPase) in bovine Leydig cells has been studied and compared with the pattern of thiamine pyrophosphatase (TPPase) and acid phosphatase distribution in these cells. Using beta-nicotinamide adenine dinucleotide phosphate (beta-NADP+) as substrate, a marked staining is observed in the intermediate Golgi saccules with some focal extension to the trans aspect. Cisternae on the cis side and associated vesicles yielded only slightly positive reactions. The pattern of NADPase localization is clearly different from that of TPPase which consistently stains only the trans Golgi elements. The specificity of NADPase for its substrate, beta-NADP+, was clearly demonstrated by using substrates modified in either the nicotinamide region e.g. alpha-nicotinamide adenine dinucleotide phosphate (alpha-NADP+), beta-thionicotinamide adenine dinucleotide phosphate (Thio-NADP+), in the attachment site of the monoester phosphate group to the molecule (e.g. 2' monophospho-adenosine 5'-diphosphoribose (ATP-ribose) or adenosine-5-monophosphate (5'AMP). With these substrates only weak or negative reactions were obtained in the Golgi apparatus of the bovine Leydig cell. PMID- 3028989 TI - Polymyxin-B-binding sites in the cochlea as demonstrated by polymyxin-B/gold labeling. AB - A polymyxin-B/bovine-serum-albumin/gold complex was used as a probe to detect the binding sites of polymyxin B on thin sections of cochlea embedded in Spurr's resin. The binding sites were found to be mainly located on the stereocilia, the cuticular plate of hair cells, the head plate of Deiters' cells, the tonofilaments in pillar cells and Deiters' cells, fibrous structures in the spiral limbus, the tectorial membrane and the basilar membrane and neural elements such as nerve endings, fibers, and the myelin sheath. The mitochondria, plasma membrane, and chromatin of the nuclei of the cells observed also exhibited binding. Our results suggest that phospholipids, glycoconjugates, cytoskeletal proteins and nucleic acids are responsible for this binding activity. PMID- 3028988 TI - Image analysis of the histochemical demonstration of glucose-6-phosphatase activity in rat liver. AB - Activities of histochemically demonstrated glucose-6-phosphatase were quantified by computerized densitometry using image analysis in livers of female adult Wistar rats fed ad libitum and fasted 22 h before sacrifice. Mean optical densities along the path between small portal tracts and efferent hepatic venous branches and enzyme activities obtained from biochemical assays exhibited a strong positive correlation. The gradients of high periportal to lower perivenous glucose-6-phosphatase activities were analysed by profiles of optical density along these distances. Mapping optical densities in an image of equidensity range provided information on the distribution pattern of hepatic glucose-6-phosphatase over an extended two-dimensional area. This visualisation of histochemical enzyme reaction based on quantitative data supports the approach of sampling across the entire protal----hepatic venous distance disregarding parenchymal zonation. Utilities provided by computer assisted image analysis will have some bearing for further adequate quantitative description of liver function and structural make up. PMID- 3028990 TI - Immunocytochemical localization of sphingolipid activator protein-1, the sulfatide/GM1 ganglioside activator, to lysosomes in human liver and colon. AB - Sphingolipid activator proteins (SAP) stimulate the enzymatic hydrolysis of sphingolipids. The results of biochemical studies have suggested that SAP are located within lysosomes. In this study we sought immunocytochemical verification of the lysosomal location of SAP-1, a SAP that stimulates the hydrolysis of sulfatide and GM1 ganglioside. We stained adjacent sections of normal adult liver and colon for either SAP-1, by peroxidase-labeled antibodies, or acid phosphatase, by enzyme histochemistry. At the light microscopic level, SAP-1 and acid phosphatase were present in similar cells of the colonic lamina propria and hepatic sinusoids, and in similar supranuclear sites of colonic epithelial cells. By electron microscopy, SAP-1 was present in vesicular structures morphologically similar to those containing acid phosphatase. Thus, SAP-1 is present in lysosomes of several different kinds of cells in the normal human liver and colon. PMID- 3028992 TI - Simple determination of erythrocyte pyrimidine 5'-nucleotidase activity in human blood by high-performance liquid chromatography. PMID- 3028991 TI - Localization of nucleotide pyrophosphatase in the rat kidney. AB - Hydrolysis of NAD by a nucleotide pyrophosphatase of renal membrane fractions has been reported previously. The aim of the present study was to localize this enzyme in the rat kidney. Nucleotide pyrophosphatase was assayed in glomeruli, in three parts of the proximal tubule and in four parts of the distal tubule dissected form freeze-dried sections. Nucleotide pyrophosphatase activity, expressed in mumol X min-1 X mg protein-1, ranged between 9.8 and 32.3 in the proximal tubular segments and between 1.1 and 2.7 in the distal tubular segments. It was 3.4 in the glomeruli. The enrichment of the activity during the purification of brush border vesicles was measured. A ten-fold higher specific activity was found in the brush border vesicles as compared to the renal cortical homogenates. Thus, most of the renal nucleotide pyrophosphatase appears to be localized in the luminal membrane of the proximal tubule. A permeabilization of the membrane did not increase the activity of brush border vesicles. This indicates that all catalytic sites are accessible at the outer surface of the membrane. PMID- 3028993 TI - Polyneuropathy caused by chronic exposure to trichloroethylene. PMID- 3028994 TI - [Gyrase inhibitor in the local treatment of the chronically infected middle ear following surgery]. AB - Local treatment of 20 chronic bacterial ear infections (18 infected radical cavities, 2 external otitis) with a gyrase inhibitor (Ciprofloxacin) solution led to clinical resolution in 17 cases, with elimination of the causative organisms in 16 cases. No side effects were seen, but a resistant pseudomonas developed in one case, and candida albicans in 8 others. Gyrase inhibitors such as Ciprofloxacin are antibiotics of second choice. For local treatment in the ear they should be used only in difficult cases and exclusively by the otologist. This treatment must not be used to replace proper cleaning of the ear or a necessary operation. PMID- 3028995 TI - Alterations of visual contrast sensitivity in Parkinson's disease. AB - Contrast sensitivity functions were determined in a population of 18 patients suffering from Parkinson's disease, and compared with the data obtained in an age matched group of healthy controls. The controls were more sensitive at all spatial frequencies tested than the patients. The statistical comparisons were highly significant, indicating general differences between the PD patients and the controls not related to individual spatial frequency channels. When comparing the sensitivity loss between low and high spatial frequencies no significant differences were found suggesting that the decrease in contrast sensitivity is a global effect. We controlled for effects of age and cerebral atrophy, and our findings cannot be accounted for by these factors. In addition, the amount of contrast sensitivity loss was not correlated with the severity of the disease. These global functional alterations appear to be related to the reduction of dopamine at various sites of the visual system. PMID- 3028998 TI - Captopril and enalapril: angiotensin-converting enzyme inhibitors. PMID- 3028997 TI - Antimicrobial chemotherapy in the adolescent patient. PMID- 3028996 TI - Accelerated radiotherapy followed by chemotherapy for locally recurrent small cell carcinoma of the lung. A phase II study of Cancer and Leukemia Group B. AB - Recurrent or persistent small-cell carcinoma of the lung (SCCL) after chemotherapy (CT) alone has shown a poor response to conventional salvage radiotherapy (RT). Accelerated RT is judged more effective than conventional RT for rapidly growing tumors such as SCCL. The objectives of this study were: to determine the tolerability of accelerated RT; and to test the ability of accelerated RT plus CT to achieve local tumor control (LTC) of SCCL recurrent after CT. Patients whose localized tumor was not controlled were selected from Arm III of the Cancer and Leukemia Group B (CALGB) protocol 8083 (Proc. ASCO 2:230, 1984) as eligible for this study. The program of accelerated RT consisted of the delivery of 50.1 Gray (Gy) in 30 fractions over a period of 21 days to the chest. New chemotherapy different from the first began 2 weeks after the completion of RT and was repeated every 3 weeks for 18 months (M). Of 29 potentially eligible patients with locally recurrent SCCL after the first line CT alone from Arm III of the CALBG protocol 8083, 12 were enrolled initially in this study. The analysis of LTC included 11 patients excluding one patient who died 4 weeks after the start of RT from liver metastases. The LTC achieved was as follow: complete remission in 8/11 (72%) and partial remission in 3/11 patients. None of the patients was converted to CR by subsequent chemotherapy. Survival ranged from 2 to 20 M, with a median survival time of 6 M. Tolerance to the subsequent CT, normal tissue reaction to accelerated RT, and the theoretical advantage of accelerated RT over conventional RT for SCCL were evaluated. PMID- 3028999 TI - Use of an inactivated vaccine for prevention of parvovirus-induced reproductive failure in gilts. AB - Gilts from dams that had been inoculated with inactivated porcine parvovirus (PPV) vaccine before breeding became seronegative to PPV by 26 weeks of age. Vaccination of these gilts with inactivated PPV vaccine at 32 weeks of age resulted in an antibody response that peaked at about 2 weeks after vaccination, with -log10 mean hemagglutination inhibiting (HI) antibody titers of less than 2. In the first-year group (82 gilts), HI titers gradually decreased, 20% of the gilts being seronegative by 6 to 7 weeks after vaccination and 75% being seronegative by 16 weeks after vaccination. In the second-year group, 93 gilts were infected naturally by a field strain of PPV at about 11 weeks after single vaccination with inactivated PPV. Additionally, in the second year, 20 vaccinated and 6 nonvaccinated gilts were immune-challenged with virulent PPV at 10 to 12 weeks after vaccination. Neither field nor challenge PPV infection of vaccinated pregnant gilts caused reproductive failure, even though some of the gilts became seronegative for PPV before challenge. Our findings suggest that single vaccination of gilts with inactivated PPV vaccine should give adequate protection from PPV-induced reproductive failure, even though serum HI titers decrease to an undetectable level shortly before PPV infection. PMID- 3029000 TI - Serologic prevalence of caprine arthritis-encephalitis virus in California goat dairies. AB - Fifty-three percent of goats in 13 California goat dairies had antibodies to caprine arthritis-encephalitis virus (CAEV), as determined by agar-gel immunodiffusion. Those goat dairies that reared kids on pasteurized milk had a lower seroprevalence than those that did not. Age, rearing kids on unpasteurized milk, and the presence of large joints were associated with antibodies to CAEV. Breed was associated with seroreactivity, but the association was confounded by other factors. Sex was not associated with antibodies to CAEV. The relationship between age and antibodies to CAEV was observed for goats reared on pasteurized or unpasteurized milk, which indicated that continued horizontal (contact) transmission may be important on these dairies and limited the effect of a pasteurized rearing program on control of CAEV infection. PMID- 3029001 TI - Sensory neuronopathy in a dog. PMID- 3029002 TI - 31P nuclear magnetic resonance analysis of lung cancer: the perchloric acid extract spectrum. AB - 31P-NMR spectra were obtained from the perchloric acid extracts of normal lung and lung cancer tissues obtained at surgery, and from extracts of neoplastic cells cultured in vitro. The perchloric acid extract of lung cancer tissue gave rise to a signal whose chemical shift was 3.2 ppm at pH 8.0. This signal was not observed in the extract of normal tissue of the lung from the same patient. The compound giving this signal was identified as phosphorylcholine. PMID- 3029003 TI - Cytogenetic effects of multiagent chemotherapy on the peripheral lymphocytes of patients with small cell lung cancer. AB - Peripheral lymphocytes of 8 patients with small cell lung cancer (SCLC), who received combination chemotherapy consisting of cisplatin and VP-16 (PVP), were examined for frequency of sister chromatid exchange (SCE) and chromosomal aberrations. Three of the 8 patients were treated with the PVP regimen only; however, the others had previously been treated with multiagent chemotherapy consisting of a cyclophosphamide, adriamycin, and vincristine (CAV) regimen or a cyclophosphamide, ACNU and vincristine (CAV') regimen with or without radiotherapy. Significantly increased SCE frequency was observed in previously untreated patients 3 or 4 days after PVP treatment, compared with the pretreatment values. The earlier analysis in the previously treated patients also showed increased SCE frequency, which seemed to return to the normal value as time elapsed after PVP. In addition, abnormal chromosome count distribution and/or increased incidence of structural changes were observed in cultures from all the patients. In general, striking changes in chromosomes were observed in previously treated patients. The aberrations observed in pretreated patients consisted of chromatid gaps and breaks, exchanges, fragments and dicentric chromosomes. In addition to these abnormalities, double minutes like microchromosomes were seen irrespective of radiotherapy. Further studies are needed to elucidate whether any of the chromosomal aberrations observed in this study could participate in the induction of secondary neoplasms. PMID- 3029004 TI - Isolation of human immunodeficiency virus from a Japanese hemophilia B patient with AIDS. AB - Human immunodeficiency virus (HIV) was isolated from a Japanese hemophilia B patient with AIDS. This isolate, HIV[GUN-1], was infectious to several mature T cell lines. Proteins with apparent molecular weights of 160, 55 and 25 kilodaltons were detected. Restriction enzyme cleavage patterns of the proviral genome indicated that HIV[GUN-1] is related to but clearly different from HTLV III or ARV-2. PMID- 3029005 TI - Inhibition of simian virus 40 replication by kanamycin derivative. PMID- 3029006 TI - Effect of feed treatment and exogenous estrogen and progestogen on puberty and lambing rates in ewe lambs. AB - Four hundred seventy-four ewe lambs (5 to 6 mo of age) were assigned within breed to two postweaning feed treatments; 1 = alfalfa pellets ad libitum and 2 = alfalfa pellets plus 20% barley or wheat ad libitum. Ten days prior to the start of breeding, approximately one-half of the ewe lambs within each feed treatment were treated with a single injection of 2.5 mg of estradiol valerate and 1.5 mg of norgestomet and implanted with 3 mg of norgestomet for 8 d. At the start of breeding, fertile Suffolk rams were fitted with marking harnesses and penned with ewe lambs; evidence of estrus and breeding was determined on a weekly basis by visual examination for breeding marks. In the second year of the study, 7 to 13 d after recording estrous activity, all marked ewe lambs were bled; and blood was assayed for progesterone. In 1983, 60% of the ewe lambs showed estrus and 11% lambed compared with 48% exhibiting estrus and 30% lambing in 1984 (P less than .01). More (P less than .01) ewe lambs on feed treatment 1 displayed estrus, but more (P less than .05) lambed in the feed treatment 2 group. Steroid treatment resulted in more (P less than .01) ewe lambs showing estrus but fewer lambing (P less than .01). Examination of progesterone concentrations for evidence of ovulation and corpus luteum development indicated that treatment with steroids caused a large percentage of ewe lambs to exhibit estrus. However, many of these failed to develop a corpus luteum and become pregnant.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3029007 TI - The effects of corn silage dry matter content and sodium bicarbonate addition on nutrient digestion and growth by lambs and calves. AB - One lamb and three calf trials were conducted to determine if interactions existed between the effects of dry matter (DM) content of corn silage at harvest and sodium bicarbonate supplementation on diet digestibility, nitrogen balance, rate and site of digestion and feedlot performance. Corn plants were harvested from the same field when DM content was approximately 31% (early; E) or 44% (late; L). Sodium bicarbonate (1.2% of DM intake) was added in a completely mixed ration. When lambs were offered diets ad libitum, (trial 1), intakes were greater (P less than .05) for L-silage diets, but apparent digestibilities were similar. Nitrogen balance was greater for sodium bicarbonate-supplemented diets, and was a reflection of greater (P less than .10) DM intakes for these diets. At similar diet DM intakes, N balance was greater for L-silage diets, with no effects measured due to sodium bicarbonate addition. When growing, abomasally cannulated heifers (trial 2) were offered diets 12 times per day, a significant interaction among treatments was measured for total tract and acid detergent fiber (ADF) digestion. Adding bicarbonate to the E-silage diet increased (P less than .05) digestion by 9.1 percentage units (66.4 vs 75.5%) but decreased digestibility of the L-silage diet by 4.2 percentage units (73 vs 68.8%). Grams of ADF apparently digested in the rumen followed the same pattern as for total tract digestion. In trial 3, ruminal rates of liquid and particulate passage and in situ rats of NDF digestion were determined using four rumen-cannulated heifers calves.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3029008 TI - Responses to dietary additions of fiber (alfalfa meal) in growing pigs housed in a cold, warm or hot thermal environment. AB - Five trials involving 112 individually-penned animals were conducted to determine the effects of dietary fiber additions (0 or 10% dehydrated alfalfa meal) on the rate and efficiency of growth and carcass characteristics of growing pigs housed in a cold (10 C), thermoneutral (22.5 C) or hot (35 C) thermal environment. Cold exposure (10 C) depressed efficiency of feed and energy utilization, carcass length, backfat, cold carcass dressing percentage and percentage lean cuts. Exposure to the hot environment resulted in depressed live weight gains but did not alter the efficiency of feed utilization. Dietary additions of alfalfa meal depressed daily gains by 1, 3 and 5% in pigs housed in the cold, warm and hot environments, respectively, and gain:feed ratios by 1, 7 and 10%. Ingestion of the fibrous alfalfa meal depressed cold carcass dressing percentage in the 10 and 22 C environments as compared with the basal diet. Based on these data, the nutritional value of fibrous feedstuffs (15% total dietary neutral detergent fiber or less) for growing pigs allowed to consume feed ad libitum is greater in animals housed in a moderately cold vs a warm or hot thermal environment. PMID- 3029009 TI - Effect of dietary phosphorus and roughage levels on calcium, magnesium and potassium utilization by sheep. AB - Three levels of dietary P (.12%, .24% and .48% of dry matter) and three levels of roughage as ground corn cobs (25% 50% and 75% of dry matter) were fed in a 3 X 3 factorial metabolism trial, utilizing 36 crossbred (Hampshire X Columbia) intact male lambs, 6 to 9 mo of age. All diets contained cane molasses (5%), blood meal (13.5%), urea (1%), corn oil (1%) and salt (.5%). Limestone supplied supplemental Ca and treatment P levels were supplied by monosodium phosphate. Equal levels of corn starch and cerelose supplied the remainder of the diet. The diet was fed ad libitum, once daily. The highest P level (.48% P) resulted in a negative (P less than .01) Ca balance (-.23 g/d vs .12 g/d for .12% P and .31 g/d for .24% P groups), and apparent digestibility (P less than .01) of Ca (1.65% vs 17.18 and 22.2% for the two respective lower P dietary levels). Blood serum concentrations of Ca and Mg decreased (P less than .01) as dietary P level was increased. Apparent digestibility of Mg was decreased (P less than .05) by the highest P level (6.9% vs 21.58% and 18.80% for the two lower levels of dietary P). Level of roughage had no effect on Ca and Mg utilization; however, the highest level (75% corn cobs) resulted in improved (P less than .05) K balance (.85 g/d vs .30 and .50 g/d for the two lower levels of roughage).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3029010 TI - Elimination of plasmids from Enterobacteriaceae by 4-quinolone derivatives. AB - Twelve 4-quinolones (cinoxacin, ciprofloxacin, enoxacin, flumequin, nalidixic acid, norfloxacin, oxolinic acid, pefloxacin, pipemidic acid, rosoxacin, and piromidic and beta-hydroxypiromidic acids) and novobiocin, were used at subinhibitory concentrations to eliminate from Escherichia coli 11 antibiotic resistance plasmids belonging to different incompatibility groups. The 12 4 quinolones were also tested for their ability to cure virulence plasmids from five species of Enterobacteriaceae. All quinolones eliminated three antibiotic resistance plasmids (R446b, R386, S-a) and one virulence plasmid (pWR105), but at a low rate. Optimal curing of antibiotic resistance plasmids was obtained in human urine. Two virulence plasmids (pWR24 and pWR110) were eliminated only by flumequin and pefloxacin. Novobiocin eliminated three antibiotic resistance plasmids (R446b, R386, pIP24). The variable and low level of plasmid loss may be explained by the induction of the recA system. In addition, the inability to eliminate certain plasmids could be due to their presence in high numbers per cell. PMID- 3029011 TI - Effects of defined medium, fetal bovine serum, and human serum on growth and chemosensitivities of human breast cancer cells in primary culture: inference for in vitro assays. AB - We compared the effects of defined medium, fetal bovine serum (FBS) and human serum (HuS) on the growth and responses to chemotherapeutic agents of human breast cancer cells in primary culture. Normal and tumor tissues were dissociated to small aggregates and single cells and seeded onto collagen-gel-coated wells in defined medium or medium supplemented with 5% FBS or 5% HuS. In all cases examined, defined medium and medium containing HuS were superior to medium containing FBS in supporting growth of both normal and tumor cell cultures. However, cultures in defined medium showed an initial cell loss. Cells from the same tumor cultured in different media varied in their responses to chemotherapeutic agents. In light of these results, medium supplemented with HuS, which promoted attachment of these cells in culture and stimulated their growth, should be the most appropriate nutrient environment for determining the effects of therapeutic agents on cells as it most closely resembles the in vivo situation. Because there were also variations in growth rates and chemosensitivities of tumor cells cultured in different human serum samples, we suggest that optimal conditions in which to culture these cells include the serum of the patient whose tumor is removed. This serum may provide host factors that influence cell growth and interact with exogenous factors. PMID- 3029012 TI - Phenotypic changes and gene expression in human colon mucosal epithelial cells upon transfection of a SV40 DNA-gpt recombinant. AB - Phenotypic changes (increased longevity, decreased growth factor requirements, altered cell surface features, growth in semisolid agarose, and SV40 T antigen expression) suggesting in vitro transformation were displayed by human normal colon mucosal epithelial cells transfected with pSV3gpt, a pBR322 recombinant containing the SV40 "early" T antigen coding region and the dominant selectable marker bacterial gene, xanthine-guanine phosphoribosyltransferase. In contrast, control cultures which received neither DNA nor the recombinant pSV2gpt (which is identical to pSV3gpt but lacks the SV40 T antigen region) were not phenotypically altered. PMID- 3029013 TI - Molecular cloning and characterization of the Streptomyces hygroscopicus alpha amylase gene. AB - We have isolated and sequenced a gene (amy) coding for alpha-amylase (EC 3.2.1.1.) from the Streptomyces hygroscopicus genome (H. Hidaka, Y. Koaze, K. Yoshida, T. Niwa, T. Shomura, and T. Niida, Die Starke 26:413-416, 1974). Amylase was purified to obtain amino acid sequence information which was used to synthesize oligonucleotide probes. amy-containing Escherichia coli cosmids identified by hybridization did not express amylase activity. Subcloning experiments indicated that amy could be expressed from the lac promoter in E. coli or from its own promoter in S. lividans. The amy nucleotide sequence indicated that it coded for a protein of 52 kilodaltons (478 amino acids). Secreted alpha-amylase contained amino- and carboxy-terminal as well as internal amino acid sequences which were consistent with the nucleotide sequence. The 30 residue leader sequence showed similarities to those found in other procaryotes. The DNA sequence 5' to the amy structural gene contained a sequence complementary to the 3'-terminal sequence of 16S rRNA of S. lividans (M. J. Bibb and S. N. Cohen, Mol. Gen. Genet. 187:265-277, 1982). The transcriptional start points of amy were determined by mung bean nuclease mapping, but the promoter of amy was not similar to the consensus sequence found in other procaryotes. PMID- 3029014 TI - Processing of the T-DNA of Agrobacterium tumefaciens generates border nicks and linear, single-stranded T-DNA. AB - Transfer and integration of a defined region (T-DNA) of the tumor-inducing (Ti) plasmid of Agrobacterium tumefaciens is essential for tumor formation. We used a physical assay to study structural changes induced in Agrobacterium T-DNA by cocultivation with plant cells. We show that nicks are introduced at unique, identical locations in each of the 24-base-pair imperfect direct repeats which flank the T-DNA and present evidence that a linear, single-stranded molecule is generated. We propose that these changes result from processing of the T-DNA for transfer and that they occur by a mechanism similar to DNA processing during conjugative DNA transfer between bacteria. PMID- 3029015 TI - In vivo D-serine deaminase transcription start sites in wild-type Escherichia coli and in dsdA promoter mutants. AB - The D-serine deaminase structural (dsdA) and regulatory (dsdC) genes are transcribed with opposite polarity from an intergenic region comprising more than 600 base pairs. The order of genes in the dsd region is supN-dsdA-dsdC-aroC-- his. The DNA sequence of the intergenic region has been slightly revised from a previously published version (E. McFall and L. Runkel, J. Bacteriol. 154:1508 1512, 1983). The dsdA gene is preceded by a long open reading frame. The dsdA in vivo transcription start sites for the wild type (base pair +1) and for three phenotypically distinct promoter constitutive mutants were determined by the S1 nuclease method. They are identical and are located about 81 base pairs upstream of the translation start site. D-Serine deaminase regulation is normal in rho mutants. Possible mechanisms for dsdA activation are discussed. PMID- 3029016 TI - Structural analysis of the Escherichia coli K-12 hisT operon by using a kanamycin resistance cassette. AB - We constructed a series of recombinant plasmids containing a kanamycin resistance (Kmr) cassette upstream from, within, and downstream from hisT, which encodes the tRNA modification enzyme pseudouridine synthase I. These Kmr insertions were then crossed directly into the bacterial chromosome. We determined growth characteristics, assayed in vivo hisT expression, and mapped in vivo hisT operon transcripts for the Kmr insertion mutants. We also analyzed polypeptides synthesized in minicells from plasmids containing Kmr cassettes. The combined results from these experiments demonstrate new features concerning the structure and expression of the complex operon that contains hisT. We show that the minimum size of the operon is approximately 3,500 base pairs and that it contains at least four genes, which are arranged in the order usg-2 (pdxB), usg-1, hisT, and dsg-1 and encode polypeptides with apparent molecular masses of 42,000, 45,000, 31,000, and 17,000 daltons, respectively. Of these genes, only the functions of usg-2 (pdxB) and hisT are known, and genetic evidence suggests that these two genes do not require usg-1 or dsg-1 for function, usg-2 (pdxB) is required for growth of bacteria on minimal medium at 37 degrees C. In contrast, the three genes at the end of the hisT operon are dispensable and form a transcription unit that is expressed from a relatively strong internal promoter. The phenotypes of the Kmr insertion mutants and results from gene expression experiments further confirm the position of the internal promoter and locate additional genetic signals in the DNA sequence around hisT. The experiments reported here also indicate several interesting properties of the Kmr cassette as a tool for probing complex operons. PMID- 3029018 TI - The 65-kilodalton antigen of Mycobacterium tuberculosis. AB - The immune response of the host to the antigens of Mycobacterium tuberculosis plays the key role in determining immunity from infection with as well as the pathogenicity of this organism. A 65-kilodalton (kDa) protein has been identified as one of the medically important antigens of M. tuberculosis. The gene encoding this antigen was isolated from a lambda gt11-M. tuberculosis recombinant DNA library using monoclonal antibodies directed against the 65-kDa antigen as the specific probes. The nucleotide sequence of this gene was determined, and a 540 amino-acid sequence was deduced. This sequence was shown to correspond to that of the 65-kDa antigen by constructing a plasmid in which this open reading frame was fused to the lacZ gene. The resulting fusion protein reacted specifically with the anti-65-kDa protein antibodies. A second long open reading frame was found downstream of the 65-kDa antigen gene which could encode a protein of 517 amino acids. This putative protein contained 29 tandemly arranged partial or complete matches to a pentapeptide sequence. PMID- 3029019 TI - Tn5-induced cytochrome mutants of Bradyrhizobium japonicum: effects of the mutations on cells grown symbiotically and in culture. AB - Two Bradyrhizobium japonicum cytochrome mutants were obtained by Tn5 mutagenesis of strain LO and were characterized in free-living cultures and in symbiosis in soybean root nodules. One mutant strain, LO501, expressed no cytochrome aa3 in culture; it had wild-type levels of succinate oxidase activity but could not oxidize NADH or N,N,N',N'-tetramethyl-p-phenylenediamine (TMPD). The cytochrome content of LO501 root nodule bacteroids was nearly identical to that of the wild type, but the mutant expressed over fourfold more bacteroid cytochrome c oxidase activity than was found in strain LO. The Tn5 insertion of the second mutant, LO505, had a pleiotropic effect; this strain was missing cytochromes c and aa3 in culture and had a diminished amount of cytochrome b as well. The oxidations of TMPD, NADH, and succinate by cultured LO505 cells were very similar to those by the cytochrome aa3 mutant LO501, supporting the conclusion that cytochromes c and aa3 are part of the same branch of the electron transport system. Nodules formed from the symbiosis of strain LO505 with soybean contained no detectable amount of leghemoglobin and had no N2 fixation activity. LO505 bacteroids were cytochrome deficient but contained nearly wild-type levels of bacteroid cytochrome c oxidase activity. The absence of leghemoglobin and the diminished bacterial cytochrome content in nodules from strain LO505 suggest that this mutant may be deficient in some aspect of heme biosynthesis. PMID- 3029017 TI - An unusual genetic link between vitamin B6 biosynthesis and tRNA pseudouridine modification in Escherichia coli K-12. AB - We characterized several unusual phenotypes caused by stable insertion mutations in a gene that is located upstream in the same operon from hisT, which encodes the tRNA modification enzyme pseudouridine synthase I. Mutants containing kanamycin resistance (Kmr) cassettes in this upstream gene, which we temporarily designated usg-2, failed to grow on minimal plus glucose medium at 37 and 42 degrees C. However, usg-2::Kmr mutants did form oddly translucent, mucoid colonies at 30 degrees C or below. Microscopic examination revealed that cells from these translucent colonies were spherical and seemed to divide equatorially. Addition of D-alanine restored the shape of the mutant cells to rods and allowed the mutants to grow slowly at 37 degrees C and above. By contrast, addition of the common L-amino acids prevented growth of the usg-2::Kmr mutants, even at 30 degrees C. Furthermore, prolonged incubation of usg-2::Kmr mutants at 37 and 42 degrees C led to the appearance of several classes of temperature-resistant pseudorevertants. Other compounds also supported growth of usg-2::Kmr mutants at 37 and 42 degrees C, including glycolaldehyde and the B6 vitamers pyridoxine and pyridoxal. This observation suggested that usg-2 was pdxB, which had been mapped near hisT. Complementation experiments confirmed that usg-2 is indeed pdxB, and inspection of the pyridoxine biosynthetic pathway suggests explanations for the unusual phenotypes of pdxB::Kmr mutants. Finally, Southern hybridization experiments showed that pdxB and hisT are closely associated in several enterobacterial species. We consider reasons for grouping pdxB and hisT together in the same complex operon and speculate that these two genes play roles in the global regulation of amino acid metabolism. PMID- 3029020 TI - Conservation of structure and location of Rhizobium meliloti and Klebsiella pneumoniae nifB genes. AB - Using transposon Tn5-mediated mutagenesis, an essential Rhizobium meliloti nitrogen fixation (nif) gene was identified and located directly downstream of the regulatory gene nifA. Maxicell and DNA sequence analysis demonstrated that the new gene is transcribed in the same direction as nifA and codes for a 54 kilodalton protein. In Klebsiella pneumoniae, the nifBQ operon is located directly downstream of a gene which is structurally and functionally homologous to the R. meliloti nifA gene. The DNA sequences of the K. pneumoniae nifB and nifQ genes (which code for 51- and 20-kilodalton proteins, respectively) were determined. The DNA sequence of the newly identified R. meliloti gene was approximately 50% homologous to the K. pneumoniae nifB gene. R. meliloti does not contain a gene homologous to nifQ directly downstream of nifB. The R. meliloti nifB product shares approximately 40% amino acid homology with the K. pneumoniae nifB product, and 10 of the 12 cysteine residues of the R. meliloti nifB product are conserved with 10 of the 17 cysteine residues of the K. pneumoniae nifB product. PMID- 3029021 TI - Genetic and structural analysis of the Rhizobium meliloti fixA, fixB, fixC, and fixX genes. AB - The fixA, fixB, fixC, and fixX genes of Rhizobium meliloti 1021 constitute an operon and are required for nitrogen fixation in alfalfa nodules. DNA homologous to the R. meliloti fixABC genes is present in all other Rhizobium and Bradyrhizobium species examined, but fixABC-homologous sequences were found in only one free-living diazotroph, Azotobacter vinelandii. To determine whether the fixABCX genes share sequence homology with any of the 17 Klebsiella pneumoniae nif genes, we determined the entire nucleotide sequence of the fixA, fixB, fixC, and fixX genes and defined four open reading frames that code for polypeptides of molecular weights 31,146, 37,786, 47,288, and 10,937, respectively. Neither DNA nor amino acid sequence homology to the R. meliloti fixA, -B, -C, and -X genes was found in the K. pneumoniae nif operon. The fixX gene contains a cluster of cysteine residues characteristic of ferredoxins and is highly homologous to an Azotobacter ferredoxin which has been shown to donate electrons to nitrogenase. The fixABC operon contains a promoter region that is highly homologous to other nifA-activated promoters. We also found a duplication of the 5' end of the fixABCX operon; a 250-bp region located 520 bp upstream of the fixABCX promoter bears more than 65% homology to the 5' end of the transcribed region, including the first 32 codons of fixA. PMID- 3029023 TI - Multiple SecA protein isoforms in Escherichia coli. AB - To define the anti-SecA-LacZ antiserum, immunoprecipitates produced with either whole anti-SecA-LacZ rabbit antiserum or affinity-purified antibodies were used to analyze nondenatured lysates of Escherichia coli. The antiserum contains antibodies that recognize different proteins. Antibody purified by preadsorption to the SecA-LacZ hybrid protein precipitated only the SecA protein from extracts. In contrast, antibody purified from the intact SecA protein precipitated several additional proteins with SecA protein. Ribosomal protein L7L12 is one of the polypeptides coprecipitated with SecA protein by antibody purified by immunoadsorption to the intact SecA protein as well as by unfractionated anti SecA-LacZ antiserum. Two-dimensional gel electrophoresis of the SecA protein immunoprecipitated by either antiserum or purified antibody indicated that the SecA protein exists in at least two, and probably four, isoforms. Only one of the SecA isoforms is present in a ribosomal preparation. PMID- 3029022 TI - Alteration of surface properties in a Tn5 mutant strain of Rhizobium trifolii 0403. AB - A symbiotically defective mutant strain of Rhizobium trifolii, UR251, was obtained by transposon Tn5 mutagenesis of R. trifolii 0403 rif and recognized by its partially ineffective (Fix +/-) phenotype on white clover plants. UR251 had a single Tn5 insertion in plasmid DNA, a wild-type plasmid pattern, and no detectable Mu DNA sequences originally present in the vector used for Tn5 mutagenesis. Agglutination by the clover lectin trifoliin A and attachment to clover root hairs was higher with UR251 than with the wild-type strain. The capsular polysaccharide (CPS) of UR251 was altered, as shown by a slower rate of CPS depolymerization with a CPS beta-lyase, PD-I; more pyruvate and less acetate and 3-hydroxybutanoate noncarbohydrate substitutions as quantitated by 1H nuclear magnetic resonance; and a higher pyruvyl transferase activity (enzymatic pyruvylation of lipid-bound saccharides). The site of increased pyruvylation in the CPS of UR251 was on the terminal galactose of the branch of the repeating oligosaccharide unit. These results show that the level of noncarbohydrate substitutions of the CPS as well as pyruvyl transferase activity are altered in R. trifolii UR251 and that trifoliin A-binding ability and clover root hair attachment are improved in this mutant strain of R. trifolii 0403 rif. PMID- 3029024 TI - Evidence that the Bacteroides thetaiotaomicron chondroitin lyase II gene is adjacent to the chondro-4-sulfatase gene and may be part of the same operon. AB - The chondroitin lyase II gene from Bacteroides thetaiotaomicron has previously been cloned in Escherichia coli on a 7.8-kilobase (kb) fragment (pA818). In E. coli, the chondroitin lyase II gene appeared to be expressed from a promoter that was about 0.5 kb from the beginning of the gene. However, when a subcloned 5-kb fragment from pA818 which contained the chondroitin lyase II gene and the promoter from which the gene is expressed in E. coli was introduced into B. thetaiotaomicron on a multicopy plasmid (pEG800), the chondroitin lyase specific activity of B. thetaiotaomicron was not altered. Further evidence that the promoter that is recognized in E. coli may not be the promoter from which the chondroitin lyase II gene is transcribed in B. thetaiotaomicron was obtained by making an insertion in the B. thetaiotaomicron chromosome at a point which is 1 kb upstream from the chondroitin lyase II gene. This insertion stopped synthesis of the chondroitin lyase II gene product, as would be predicted if the gene was part of an operon and was transcribed in B. thetaiotaomicron from a promoter that was at least 1 kb upstream from the chondroitin lyase II gene. A region of pA818 which was adjacent to the chondroitin lyase II gene and which included the region used to make the insertional mutation was found to code for chondro-4-sulfatase, an enzyme that breaks down one of the products of the chondroitin lyase reaction. The upstream insertion mutant of B. thetaiotaomicron which stopped synthesis of chondroitin lyase II had no detectable chondro-4-sulfatase activity. This mutant was still able to grow on chondroitin sulfate, although the rate of growth was slower than that of the wild type. PMID- 3029025 TI - Plasmid marker rescue transformation proceeds by breakage-reunion in Bacillus subtilis. AB - Bacillus subtilis carrying a plasmid which replicates with a copy number of about 1 was transformed with linearized homologous plasmid DNA labeled with the heavy isotopes 2H and 15N, in the presence of 32Pi and 6-(p-hydroxyphenylazo)-uracil to inhibit DNA replication. Plasmid DNA was isolated from the transformed culture and fractionated in cesium chloride density gradients. The distribution of total and donor plasmid DNA was examined, using specific hybridization probes. The synthesis of new DNA, associated with the integration of donor moiety, was also monitored. Donor-specific sequences were present at a density intermediate between that of light and hybrid DNA. This recombinant DNA represented 1.4% of total plasmid DNA. The latter value corresponded well with the transforming activity (1.7%) obtained for the donor marker. Newly synthesized material associated with plasmid DNA at the recombinant density amounted to a minor portion of the recombinant plasmid DNA. These data suggest that, like chromosomal transformation, plasmid marker rescue transformation does not require replication for the integration of donor markers and, also like chromosomal transformation, proceeds by a breakage-reunion mechanism. The extent of donor DNA replacement of recipient DNA per plasmid molecule of 54 kilobases (27 kilobase pairs) was estimated as 16 kilobases. PMID- 3029026 TI - Hexuronate catabolism in Erwinia chrysanthemi. AB - In the phytopathogenic enterobacterium Erwinia chrysanthemi, the catabolism of hexuronates is linked to the degradation of pectic polymers. We isolated Mu lac insertions in each gene of the hexuronate pathway and used genetic fusions with lacZ (the beta-galactosidase gene of Escherichia coli) to study the regulation of this pathway. Three independent regulatory genes (exuR, uxuR, and kdgR) were found. Galacturonate and glucuronate were converted into 2-keto-3-deoxygluconate (KDG) by separate three-step pathways encoded by the uxaC, uxaB, and uxaA genes and the uxaC, uxuB, and uxuA genes, respectively. The two aldohexuronates entered the cell by a specific transport system, encoded by exuT. Wild-type strain 3937 was unable to use glucuronate as a carbon source since glucuronate was unable to induce the exuT expression. Mutants able to use glucuronate possessed an inactivated exuR gene. The product of the regulatory gene exuR negatively controlled the expression of exuT, uxaC, uxaB, and uxaA, which was inducible in the presence of galacturonate. The two genes specifically involved in glucuronate catabolism, uxuA and uxuB, formed two independent transcriptional units regulated separately, uxuB expression was not inducible, whereas uxuA expression was induced in the presence of glucuronate and controlled by the uxuR product. KDG, the common end product of both pathways, is cleaved by the kdgK and kdgA gene products. KDG enters the cell by a specific transport system, encoded by kdgT. The regulatory gene kdgR controlled the expression of kdgT, kdgK, and kdgA and partially that of the pel genes encoding pectate-lyases. The real inducer of pectate-lyase synthesis, originating from catabolism of galacturonate or glucuronate, appeared to be KDG. The genes of E. chrysanthemi affecting hexuronate catabolism are separated into six independent transcriptional units exuT, uxaCBA, uxuA, uxuB, kdgK, and kdgA, but only three gene clusters were localized on the genetic map: exuT-uxaCBA, uxuA-uxuB-kdgK, and kdgA-exuR. PMID- 3029027 TI - Cloning and characterization of the 5' region of the cell wall protein gene operon in Bacillus brevis 47. AB - Bacillus brevis 47 secretes vast amounts of proteins derived from both middle wall protein (MWP) and outer wall protein into the medium. The 5' region of the cell wall protein gene operon was cloned into Bacillus subtilis and subsequently into B. brevis 47. On the basis of the nucleotide sequence analysis, an open reading frame coding for MWP was identified on the cloned DNA fragment. Two potential translation initiation sites for the MWP gene are located tandemly in the same reading frame. Each of the sites contains a sequence highly homologous to the 3' end of B. brevis rRNA and an initiation codon. The translational fusion of the 5' region of the MWP gene with the Bacillus licheniformis alpha-amylase gene resulted in the efficient expression of the alpha-amylase gene in B. brevis 47. Of the two potential translation initiation sites, the one located upstream could be eliminated without affecting the expression of the MWP-alpha-amylase fusion gene, suggesting that MWP is synthesized in a precursor form with a signal peptide of 23 amino acid residues. S1 nuclease mapping of the cell wall protein gene transcripts suggested the possibility of the existence of several promoters in the 5' region within 300 base pairs from the translation initiation sites; one promoter was definitely localized within this part of the 5' region, and it was capable of expressing a heterologous gene fusion at a high level. The roles of the apparent structural complexity of the 5' region of the cell wall protein gene operon are discussed in connection with the efficient gene expression. PMID- 3029028 TI - Chemotaxis in Escherichia coli: construction and properties of lambda tsr transducing phage. AB - The tsr gene of Escherichia coli, located at approximately 99 min on the chromosomal map, encodes a methyl-accepting protein that serves as the chemoreceptor and signal transducer for chemotactic responses to serine and several repellents. To determine whether any other chemotaxis or motility genes were located in the tsr region, we constructed and characterized two lambda tsr transducing phages that each contain about 12 kilobases of chromosomal material adjacent to tsr. lambda tsr70 carries sequences from the promoter-proximal side of tsr; lambda tsr72 carries sequences from the promoter-distal side of tsr. Restriction maps of the bacterial inserts in these phages and Southern hybridization analyses of the bacterial chromosome indicated that the tsr gene is transcribed in the counterclockwise direction on the genetic map. Insert deletions were isolated in lambda tsr70 and transferred into the host chromosome to examine the null phenotype of tsr. All such strains exhibited wild-type swimming patterns and chemotactic responses to a variety of stimuli, but were specifically defective in serine taxis and other Tsr-mediated responses. In addition, UV programming experiments demonstrated that Tsr and several of its presumptive degradation products were the only bacterial proteins encoded by lambda tsr70 and lambda tsr72 that required host FlbB/FlaI function for expression. These findings indicate that there are probably no other chemotaxis related genes in the tsr region. A series of tsr point mutations were isolated by propagating lambda tsr70 on a mutD host and used to construct a fine-structure map of the tsr locus. These mutations should prove valuable in exploring structure-function relationships in the Tsr transducer. PMID- 3029029 TI - Influence of GATC sequences on Escherichia coli DNA mismatch repair in vitro. AB - The effect of the number and position of DNA adenine methylation (dam) sites, i.e., d(GATC) sequences, on mismatch repair in Escherichia coli was investigated. The efficiency of repair was measured in an in vitro assay which used an f1 heteroduplex containing a G/T mismatch within the single EcoRI site. Both an increase in the number of dam sites and a shortened distance between dam site and mismatched site increased the efficiency of mismatch repair. The sequences adjacent to d(GATC) also affected the efficiency of methylation-directed mismatch repair. Furthermore, heteroduplexes with one extra dam site located close to either the 5' or 3' end of the excised base increased the repair efficiency to about the same extent. The findings suggest that the mismatch repair pathway has no preferred polarity. PMID- 3029030 TI - Analysis of the regulation of gene expression during Bacillus subtilis sporulation by manipulation of the copy number of spo-lacZ fusions. AB - The control of expression of the Bacillus subtilis spoIIA locus was analyzed by titrating gene expression against gene copy number. A plasmid integrated into the B. subtilis chromosome and carrying the spoIIA control region fused to Escherichia coli lacZ was forced to form tandem repeats by the selection of clones that grow on high levels of chloramphenicol, the antibiotic against which the plasmid determines resistance. DNA from the clones was digested with BglII, which did not cut in the reiterated region, and the size of the fragment was determined by orthogonal-field-alternation gel electrophoresis to determine the copy number. Most clones had fairly homogeneous copy numbers. Gene expression was monitored by beta-galactosidase activity. The results indicate that spoIIA was under positive control by a moiety present at about five copies per chromosome. Spore formation was not affected by amplification, so spoIIA-lacZ reiteration did not sequester a molecule required elsewhere for sporulation. PMID- 3029031 TI - Fusions of the Escherichia coli gyrA and gyrB control regions to the galactokinase gene are inducible by coumermycin treatment. AB - We have previously shown that the genes encoding the two subunits of Escherichia coli DNA gyrase are regulated in a manner which is dependent on DNA conformation. When the DNA encoding the gyrA and gyrB genes is relaxed, both genes are expressed at a high level; in negatively supercoiled DNA they are expressed at a low level. In this paper we describe fusions of both the gyrA and gyrB 5' sequences to the E. coli galactokinase gene. In such fusions we found that galactokinase can be induced by treating the cells with coumermycin A1, an inhibitor of DNA gyrase. Our results suggest that the regulation occurs at the transcriptional level and that only a small region of DNA is necessary for coumermycin-induced gene expression. PMID- 3029032 TI - secD, a new gene involved in protein export in Escherichia coli. AB - New mutants of Escherichia coli altered in protein export were identified in phoA lacZ and lamB-lacZ gene fusion strains by searching for mutants that showed an altered lactose phenotype. Several mutations mapped in a new gene, secD. These mutants were, in general, cold sensitive for growth, and the mutations led to an accumulation of precursor of exported proteins. The secD gene is closely linked to tsx on the E. coli chromosome, but separable from another gene proposed to be involved in export, ssaD, which maps nearby. A plasmid carrying secD+ was identified and used to show that the mutations are recessive. The secD gene may code for a component of the cellular export machinery. PMID- 3029033 TI - Tn916-induced mutations in the hemolysin determinant affecting virulence of Listeria monocytogenes. AB - A genetic determinant essential for hemolysin production by Listeria monocytogenes has been inactivated by insertion of transposon Tn916 into L. monocytogenes DNA. The transposon was transferred by means of conjugation of a streptomycin-resistant L. monocytogenes recipient strain with Streptococcus faecalis CG110 on membrane filters. Among the tetracycline-resistant transconjugants, mutants were detected which had lost hemolytic activity. When tested in a mouse model, these mutants appeared to have lost the virulence that characterizes the parental strain. An extracellular protein of 58,000 apparent molecular weight was eliminated in the nonhemolytic mutants. In some of the mutants, the decrease in the production of the 58,000-dalton protein was accompanied by the production of a new protein of 49,000 apparent molecular weight. Hemolytic revertants regained the hemolytic phenotype and virulence and produced the extracellular protein that characterizes the recipient strain. Hybridization studies with Tn916 DNA indicated that the transposon is present in EcoRI and HindIII fragments of the nonhemolytic mutants. Single copies of Tn916 were detected in the chromosomal DNA of two of the three nonhemolytic mutants that were studied in detail. In hemolytic, tetracycline-sensitive revertants Tn916 appeared to be completely excised from the chromosome. PMID- 3029034 TI - Tn5 insertion in the polynucleotide phosphorylase (pnp) gene in Escherichia coli increases susceptibility to antibiotics. AB - A Tn5 insertional mutation on the Escherichia coli chromosome which caused a severalfold increase in susceptibility to structurally and functionally diverse antibiotics was found to map within the gene for polynucleotide phosphorylase (pnp) and to inactivate this enzyme, which is involved in RNA breakdown. The mutation also decreased the growth rate 10 to 25% and increased the rate of tetracycline uptake about 30%. The hypersensitivity due to the insertion was only partially complemented by a cloned pnp gene. PMID- 3029035 TI - Characterization of suppressible mutations in the viomycin phosphotransferase gene of the Streptomyces enteric plasmid pVE138. AB - The viomycin phosphotransferase gene (vph) is expressed and confers resistance to viomycin in both Streptomyces spp. and members of the family Enterobacteriaceae. We report the isolation of UGA (opal) and UAG (amber) mutations in the vph gene of shuttle plasmid pVE138. We found that the five UGA mutations in vph resulted in a temperature-sensitive phenotype in Salmonella typhimurium. Su- strains are Vior at 28 degrees C and Vios at 37 degrees C, whereas Su+UGA strains are Vior at both 28 and 37 degrees C. The single amber mutation isolated was not temperature sensitive and resulted in the expected Vios phenotype in Su- strains and Vior in Su+UAG strains. PMID- 3029036 TI - N4-methylcytosine as a minor base in bacterial DNA. AB - The DNA base composition, including the minor base content, of 26 strains of bacteria was determined. The studied bacteria are sources of widely used restriction endonucleases. Approximately 35% of the bacterial DNAs contained N4 methylcytosine, about 60% contained 5-methylcytosine, and about 90% had N6 methyladenine. PMID- 3029038 TI - Cloning and expression in Escherichia coli of a Klebsiella ozaenae plasmid-borne gene encoding a nitrilase specific for the herbicide bromoxynil. AB - An enzyme (nitrilase) that converts the herbicide bromoxynil (3,5-dibromo-4 hydroxybenzonitrile) to its metabolite 3,5-dibromo-4-hydroxybenzoic acid was shown to be plasmid encoded in the natural soil isolate Klebsiella ozaenae. The bromoxynil-specific nitrilase was expressed in Escherichia coli by direct transfer and stable maintenance in E. coli of a naturally occurring 82-kilobase K. ozaenae plasmid. Irreversible loss of the ability to metabolize bromoxynil both in E. coli and K. ozaenae was associated with the conversion of the 82 kilobase plasmid to a 68-kilobase species. In E. coli this conversion was the result of a host recA+-dependent recombinational event. A gene, designated bxn, encoding the bromoxynil-specific nitrilase was constitutively expressed in K. ozaenae and E. coli and subcloned on a 2.6-kilobase PstI DNA segment. The polarity and the location of the gene were determined by assaying hybrid constructs of the bromoxynil-specific nitrilase gene fused with the heterologous lac promoter. PMID- 3029037 TI - Promoter and nucleotide sequences of the Zymomonas mobilis pyruvate decarboxylase. AB - DNA sequence analysis showed that pyruvate decarboxylase (one of the most abundant proteins in Zymomonas mobilis) contains 559 amino acids. The promoter for the gene encoding pyruvate decarboxylase was not recognized by Escherichia coli, although the cloned gene was expressed at relatively high levels under the control of alternative promoters. The promoter region did not contain sequences which could be identified as being homologous to the generalized promoter structure for E. coli. Hydropathy plots for the amino acid sequence indicated that pyruvate decarboxylase contains a large number of hydrophobic domains which may contribute to the thermal stability of this enzyme. PMID- 3029039 TI - Isolation and characterization of an aminolevulinate-requiring Rhodobacter capsulatus mutant. AB - Using transposon Tn5 mutagenesis, we isolated a mutant strain of Rhodobacter capsulatus that requires aminolevulinate for growth. Southern blot analysis indicated that this strain has a single Tn5 insertion. The addition of 0.1 mM aminolevulinate to the medium allowed the mutant to grow either aerobically or photosynthetically with generation times similar to those of the parental strain. When grown photosynthetically, bacteriochlorophyll accumulation increased with increasing aminolevulinate concentration. The mutant strain had only 10% of the normal aminolevulinate synthase activity, but it had a normal level of porphobilinogen synthase activity. The requirement for aminolevulinate could be satisfied by porphobilinogen, hemin, or protoporphyrin. While the mutant grew well on agar plates containing any of these substrates, growth in liquid media containing hemin or protoporphyrin was poor. Introduction of an R' factor containing all the known R. capsulatus bch genes into the mutant strain did not relieve the requirement for aminolevulinate, suggesting that the Tn5 insertion is not within the bch region. PMID- 3029040 TI - The mRNA for an inducible chloramphenicol acetyltransferase gene is cleaved into discrete fragments in Bacillus subtilis. AB - cat-86 is a promoter-deficient plasmid gene that encodes chloramphenicol acetyltransferase (CAT). Insertion of a promoter at a site 144 base pairs 5' to the cat-86 coding sequence activates transcription of the gene and allows cat-86 to specify chloramphenicol-inducible CAT activity in Bacillus subtilis. Induction of cat-86 by chloramphenicol has been shown to result from a regulatory event that activates translation of cat-86 mRNA that is present in cells before the addition of inducer (E. J. Duvall and P. S. Lovett, Proc. Natl. Acad. Sci. USA 83:3939-3943, 1986). In the present study we show an unusual property of cat-86 mRNA. Full-length cat-86 transcripts, consisting of 920 nucleotides (nt), are cleaved in B. subtilis to yield two predominant fragmentation products: an 810-nt species that lacks sequences present at the 5' end of the 920-nt species and a 720-nt species that lacks sequences present at the 3' end of the 920-nt species. A third fragmentation product consisting of 620 nt may result from the cleavage of a single 920-nt transcript at both the 5' and 3' ends. The sequences which are missing from the 720- and 620-nt species suggest that these transcripts cannot be translated into functional CAT. The 810-nt species lacks sequences from the 5' regulatory region, and it is not yet certain whether or not translation of this species can be induced by chloramphenicol. The ratio of 920-nt molecules/720-nt molecules in rifampin-treated cells is increased when the cells are grown in chloramphenicol. Therefore, induction may partially stabilize full-length cat-86 transcripts against inactivation by a novel processing-like system. PMID- 3029041 TI - Capsule synthesis in Escherichia coli K-12 is regulated by proteolysis. AB - lon mutants of Escherichia coli K-12 are defective in an ATP-dependent protease, are UV sensitive, and overproduce the capsular polysaccharide colanic acid. Six structural genes needed for capsular polysaccharide synthesis (cps) are transcriptionally regulated by lon as well as by three other regulatory genes, rcsA, -B, and -C (S. Gottesman, P. Trisler, and A. S. Torres-Cabassa, J. Bacteriol. 162:1111-1119, 1985). We have cloned rcsA, the gene for a positive regulator of capsule synthesis, onto multicopy plasmids and defined the gene by both insertions and deletions. The product of rcsA has been identified as an unstable protein of 27 kilodaltons. RcsA has a half-life of 5 min in lon+ cells and one of 20 min in lon cells. The availability of RcsA is the limiting factor for capsule synthesis; doubling the gene dosage of rcsA+ significantly increases expression of cps genes. Our results are consistent with a model in which the presence of a lon mutation increases the synthesis of capsular polysaccharide via stabilization of RcsA. PMID- 3029042 TI - Constitutive expression of tetracycline resistance mediated by a Tn10-like element in Haemophilus parainfluenzae results from a mutation in the repressor gene. AB - The Tn10-like constitutively expressed tetracycline resistance determinant from a Haemophilus parainfluenzae strain was cloned in Escherichia coli. Toxicity resulting from expression on multicopy plasmids necessitated its being cloned on a low-copy plasmid vector or in cells containing the Tn10-encoded repressor. Constitutive expression of tetracycline resistance was found to result from the synthesis of a truncated inactive repressor molecule. Instead of the 23 kilodalton repressor found in other Tn10-containing strains, this determinant encoded a 14.5-kilodalton molecule. The DNA sequence of the 700-base-pair region spanning the repressor gene and promoter-operator regions of the Haemophilus determinant was identical to that of the same region of Tn10, except for the absence of a single T X A base pair in the repressor gene. This deletion leads to premature termination of the protein. Antisera to the repressor suggested that the repressor was also absent in a second independently isolated H. parainfluenzae strain bearing a Tn10-like constitutive tetracycline resistance determinant. PMID- 3029043 TI - Bacillus subtilis rRNA promoters are growth rate regulated in Escherichia coli. AB - rRNA promoters from the rrnB locus of Bacillus subtilis and from the rrnB locus of Escherichia coli were fused to the gene for chloramphenicol acetyltransferase (CAT). The level of expression of CAT in E. coli showed growth rate dependence when the CAT gene was linked to either E. coli or B. subtilis tandem promoters. The downstream promoter of the tandem Bacillus pair showed growth rate regulation, while the upstream promoter did not, whereas for the E. coli tandem promoters, only the upstream promoter was growth rate regulated. PMID- 3029045 TI - Receptor alterations associated with serotonergic agents: an autoradiographic analysis. AB - Controversy exists concerning whether receptor down-regulation is involved in the efficacy of antidepressants. Many investigators believe that norepinephrine (NE) receptor down-regulation is more important than serotonin (5-HT) receptor down regulation. The ability to accurately determine which receptor types or subtypes have been down-regulated has been impaired by the lack of sufficiently specific ligands for labeling these receptor subtypes. Studies that have attempted to examine 5-HT2 receptor down-regulation have used [3H]-ketanserin as the ligand of choice to label 5-HT2 receptors, but this ligand also labels a nondescript site. The binding of [3H]-ketanserin to sites other than 5-HT2 receptors can be examined and controlled for by autoradiographic techniques. The authors briefly review potential problems involved in analyzing receptor binding after antidepressant treatment and present new findings of receptor alterations in rat brain as examined by autoradiographic techniques following chronic exposure to fluoxetine (a selective 5-HT uptake inhibitor that has been shown to be an effective antidepressant). Laboratory animals injected with fluoxetine showed receptor down-regulation (reduced density) in the serotonergic system. A provocative and potentially important finding of this study is that this selective 5-HT uptake blocker also down-regulates beta-adrenergic receptors in the CNS. PMID- 3029044 TI - Serotonin-norepinephrine receptor interactions in the brain: implications for the pharmacology and pathophysiology of affective disorders. AB - When chronically administered, most clinically effective antidepressant treatments (pharmacotherapy and ECT) reduce the sensitivity of the norepinephrine sensitive adenylate cyclase in brain which, in turn, is associated with a down regulation of the beta-adrenoceptor subpopulation. Because this norepinephrine receptor system is linked to an amplifier system, small changes in the number of receptors or in the accumulation of the second messenger cyclic AMP will be amplified. Results of the studies discussed in this paper demonstrate that an intact serotonergic neuronal input is required for the proper functioning of beta adrenoceptors and for the down-regulation of the density of these receptors by antidepressant treatments. Under conditions of impaired serotonergic activity, beta-adrenoceptors display profound decreases in agonist but not in antagonist affinity. The changes are reminiscent of "uncoupled" receptors. While beta adrenoceptors are coupled in a stimulatory fashion to adenylate cyclase, resulting in the formation of the second messenger cyclic AMP, serotonin (5-HT) receptors are linked to phosphatidylinositol hydrolysis (5-HT2 receptors in cortex, 5-HT1C receptors in choroid plexus) generating two second messengers, diacylglycerol and inositol-1,4,5-trisphosphate. The final common pathway of aminergic receptor activation seems to be protein-kinase-mediated protein phosphorylation leading to changes in cellular activity. Evidence is presented suggesting that the delayed down-regulation of the linked 5-HT/norepinephrine beta-adrenoceptor system by antidepressant treatment reflects a therapeutically relevant biochemical action and prompts the generation of the "5 HT/norepinephrine link hypothesis" of affective disorders. PMID- 3029046 TI - Glycosphingolipids of normal bovine and enzootic bovine leukosis lymph node cells. AB - We analyzed glycosphingolipids from normal lymph node cells of seven cattle and lymph node cells of eight cattle with enzootic bovine leukosis. The neutral glycosphingolipids and gangliosides were analyzed by thin-layer chromatography. Both normal and tumorous lymph node cells had GlcCer, LacCer, and GbOse3Cer as major neutral glycosphingolipids. In the ganglioside fraction, GM3 was the predominant component in both normal and tumorous lymph node cells, and another component, ganglioside Gx fraction, was also prominent in tumorous lymph node cells. The structure of this ganglioside Gx fraction was elucidated by thin-layer chromatography, sugar analysis, neuraminidase digestion, and permethylation studies. This ganglioside Gx fraction was found to be a mixture of four ganglioside species. The structures of individual gangliosides Gx (1 to 4) were characterized as follows. 1: GD3, NeuAc alpha 2-8NeuAc alpha 2-3Gal1-4Glc-Cer. 2: GD3, NeuAc alpha 2-8NeuGc alpha 2-3Gal1-4Glc-Cer. 3: GD3, NeuGc alpha 2-8NeuAc alpha 2-3Gal1-Glc-Cer. 4: GD3, NeuGc alpha 2-8NeuGc alpha 2-3Gal1-4Glc-Cer. These GD3 species may be formed as a result of the induced synthesis inassociation with malignant transformation. PMID- 3029047 TI - Autonomous replicating sequences from intron of human ras gene in a simian virus 40 T antigen dependent system. AB - Of the several DNA fragments present in the human lung cancer gene, 1.1 and 2.0 kilobase (kb) fragments corresponding to the intron of this gene were hybridized to a half part of the 27 nucleotides perfect palindrome present in the initiation part of replication in simian virus 40 (SV40) DNA. These two fragments cloned in pBR322 had good template activity, and the initiation of DNA replication started from the region of these fragments in an in vitro system, in which the initiation of DNA replication occurs on cloned DNA containing SV40 origin of DNA replication as described previously. Furthermore, these two clones could replicate autonomously in nuclei of SV40 transformed Cos cells, producing SV40 T antigen constitutively when the clones were transfected into Cos cells. These results show that functional SV40 origin-like sequences are present in human genomes, and they can replicate autonomously within the cells which are producing SV40 T antigen. PMID- 3029048 TI - Isolation and characterization of the complete complementary and genomic DNA sequences of human serum amyloid P component. AB - Complementary and genomic DNA clones corresponding to the human serum amyloid P component (SAP) mRNA have been isolated and analyzed. The nucleotide sequences of the cDNA and the corresponding regions of the genomic SAP DNA reported here were identical, and revealed that after coding for a signal peptide of 19 amino acids and the first two amino acids of the mature SAP protein, there is one small intron of 115-base pairs (bp), followed by a nucleotide sequence coding for the remaining 202 amino acid residues. The SAP gene has an ATATAAA sequence 29-bp upstream from the cap site, but there is no CAAT box-like sequence. A possible polyadenylation signal sequence, ATTAAA, was found to be located 28-bp upstream from the polyadenylation site. A comparison of the genomic SAP DNA sequence with that of human C-reactive protein (CRP) revealed a striking overall homology which was not uniform: several highly conserved regions were bounded by non-homologous regions. This comparison provides further support for the hypothesis that SAP and CRP are products of a gene duplication event. PMID- 3029049 TI - Distinct occurrence of phosphatidylinositol 4,5-bisphosphate-induced Ca2+ release and inositol 1,4,5-triphosphate-induced release in ATP-dependent Ca2+ transporting platelet microsomes. AB - The effects of phosphatidylinositol 4,5-bisphosphate (PtdInsP2) and inositol 1,4,5-triphosphate(InsP3) on the Ca2+ release from ATP-dependent Ca2+ transporting microsomes prepared from ox platelets were investigated. Under optimal conditions, both PtdInsP2 and InsP3 released Ca2+ from the microsomes in a similar dose-dependent manner. However, the maximal amount of Ca2+ released by InsP3 was almost one-fourth of that released by PtdInsP2. Neither PtdInsP2 nor InsP3 appeared to act as a Ca2+ ionophore since they showed no effect on the Ca2+ content of liposomes prepared from platelet microsomal lipids. InsP3-induced but not PtdInsP2-induced Ca2+ release was decreased with increasing extravesicular Ca2+ from 0.1 microM to 10 microM and it was completely inhibited by 10 microM Ca2+. PtdInsP2-induced but not InsP3-induced Ca2+ release was markedly inhibited by Mg2+, ruthenium red and neomycin. In addition, InsP3 could induce no additional Ca2+ release after the accumulated Ca2+ had been maximally released by PtdInsP2. These results indicate that PtdInsP2 releases Ca2+ from platelet microsomes more effectively than InsP3 by a mechanism distinct from that of InsP3 induced release, and further that InsP3-sensitive microsomes are included within the population of PtdInsP2-sensitive microsomes. PMID- 3029050 TI - Molecular cloning and sequence analysis of full-length cDNA for rabbit liver NADPH-cytochrome P-450 reductase mRNA. AB - The nucleotide sequence of the mRNA for NADPH-cytochrome P-450 reductase from rabbit liver was determined from a full-length cDNA clone (pFP105). The clone contains 2,269 nucleotides complementary to rabbit liver reductase mRNA. The single open reading frame of 2,037 nucleotides codes for a 679-amino acid polypeptide with a calculated molecular weight of 76,583 daltons. The cloned cDNA contains the complete 3'-noncoding region of 193 nucleotides, including 68 nucleotides of poly(A), and 39 nucleotides of the 5'-noncoding region. The nucleotide sequence in the coding region of cDNA of rabbit reductase (pFP105) showed 85% homology to that of rat reductase (Porter, T.D. & Kasper, C.B. (1985) Proc. Natl. Acad. Sci. U.S. 82, 973-977, and Murakami, H. et al. (1986) DNA 5, 1 10). Rabbit reductase has one more amino acid residue than the rat enzyme, and the amino acid compositions of the two enzymes are similar. The amino acid sequence of the rabbit enzyme showed 91% identity with that of the rat enzyme. The segment related to binding of FMN and FAD was well conserved among rabbit, rat, and pig reductases. The sequence related to AMP moiety-binding was also conserved among these species, and was found in the amino acid sequence of NADH cytochrome b5 reductase, another flavoenzyme in the microsomal electron transport system. PMID- 3029051 TI - Developmental variation and amino acid sequences of cytochromes c of the fruit fly Drosophila melanogaster and the flesh fly Boettcherisca peregrina. AB - The amino acid sequences of cytochromes c purified from the fruit fly Drosophila melanogaster and the flesh fly Boettcherisca peregrina were determined. In contrast with the case of the housefly, isocytochromes c were not detected in these flies at any developmental stage. The sequence of fruit fly cytochrome c differed from that reported previously but was identical with that predicted from the nucleotide sequence of the fruit fly cytochrome c gene (DC4) (Limbach, K.J. & Wu, R. (1985) Nucl. Acids Res. 13, 631-644). Isocytochrome c of the fruit fly, reported to be encoded by the DC3 gene, was not detected as a functional cytochrome c molecule. PMID- 3029052 TI - Autoxidizability of beef heart cytochrome c1 lacking the hinge protein c1-c. AB - The autoxidizability of beef heart cytochrome c1 was investigated in terms of the integrity of the binding of the hinge protein to the heme subunit. Cytochrome c1 was isolated as a subcomplex consisting of the heme subunit and the hinge protein. Treatment of the cytochrome c1 subcomplex with p-chloromercuribenzoate (pCMB) under mild conditions lessened the binding strength between the two subunits. They were dissociated on polyacrylamide gel electrophoresis (PAGE) under nondenaturing conditions, but were not separated by gel filtration chromatography. The pCMB-treated subcomplex had a slight autoxidizability. This was repressed to the level of the native subcomplex, when the mercurial compound bound to the subcomplex was removed by the addition of 2-mercaptoethanol. Concomitantly, the less stable binding between the subunits was apparently reversed to the native state. After pCMB treatment of the subcomplex, the heme subunit recovered from PAGE showed marked autoxidizability, even if it was treated with 2-mercaptoethanol. Addition of cholate repressed the autoxidizability of the heme subunit after the removal of the mercurial compound. These results confirmed that the stable binding of the hinge protein to the heme subunit was essential for the nonautoxidizability of cytochrome c1 subcomplex. In addition, it was suggested that cysteinyl residues in the subcomplex must be involved to a great extent in the stable binding between the two subunits. PMID- 3029053 TI - Acceleration of the rate of fluorescence decrease by high concentrations of ATP under the condition of accumulation of ADP-sensitive phosphoenzyme in Na+,K+ ATPase. AB - Addition of up to 300 microM ATP in the presence of 2 M NaCl with MgCl2 to pig kidney Na+,K+-ATPase treated with N-[p-(2-benzimidazolyl)phenyl]maleimide seemed to be insufficient to saturate the rate of the fluorescence decrease. However, both the extent of the decrease and the amount of phosphoenzyme at a steady state were saturated below 20 microM ATP. Addition of Mg2+ with Na+ to the enzyme preincubated with 20 to 600 microM ATP gave nearly the same rate constant, which was below 50% of that obtained by adding 300 microM ATP to the Na+-form enzyme in the presence of Mg2+. High concentrations of ATP affected neither the rate of light-scattering change (Taniguchi, K. et al. (1986) J. Biol. Chem. 261, 3272 3281) after ADP-sensitive phosphoenzyme formation (E1P) nor that of the breakdown of E1P. A stoichiometric amount of [32P]Pi was liberated from [32P]E1P. The data suggested that ATP did not bind to E1P in such a way as to increase the extent of phosphorylation further or to accelerate dephosphorylation. The data also suggested that the reason for the large difference in the apparent affinity of ATP as evaluated from the rate and the extent of fluorescence change is the large dissociation constant for ATP of a Michaelis complex. PMID- 3029054 TI - DNA methylation and the regulation of aldolase B gene expression. AB - DNA methylation was studied as a potential factor for the regulation of tissue specific and developmentally specific expression of the rat aldolase B gene. We examined cytosine methylation in the HpaII and HhaI recognition sequences in the aldolase B gene in aldolase expressing and nonexpressing tissues and cells. Out of the 15 methyl-sensitive restriction sites examined, the sites in the 3'-half and 3'-flanking regions were found to be heavily methylated in all the tissues or cells, regardless of the level of aldolase B gene expression. However, the methylation pattern in the region immediately upstream and in the 5'-half of the gene exhibited tissue-specificity: the site located about 0.13 kb upstream of the cap site (just next to the CCAAT box), and the sites in the first intron (intron 1) were heavily methylated in nonexpressing cells and tissues (ascites hepatoma AH130 and brain), whereas those in an expressing tissue (liver) were considerably less methylated. These results suggest that cytosine methylation at the specific sites in the 5'-flanking and 5'-half regions of the gene is associated with repression of the gene activity. However, the gene is still substantially methylated in the fetal liver on day 16 of gestation, when it is in a committed state for rapid activation in the period immediately afterwards (Numazaki et al. (1984) Eur. J. Biochem. 152, 165-170). This suggests that demethylation of the methylated cytosine residues in the specific gene region is not necessarily required before activation of the gene during development, but it may occur along with or after the activation. PMID- 3029055 TI - ATP depletion causes a reversible redistribution and inactivation of a subpopulation of galactosyl receptors in isolated rat hepatocytes. AB - Isolated rat hepatocytes, treated with metabolic energy poisons such as NaN3 in the absence of exogenous ligand, lose surface galactosyl (Gal) receptor activity (Clarke, B. L., and Weigel, P. H. (1985) J. Biol. Chem. 260, 128-133). We have used 125I-labeled asialo-orosomucoid and affinity-purified anti-receptor IgG to quantitate, respectively, the activity and the amount of Gal receptor protein. Cells were treated with NaN3 at 37 degrees C and the surface or total (surface and intracellular) binding of these two probes was measured at 4 degrees C, respectively, in intact cells or in cells permeabilized with digitonin. As a function of NaN3 concentration, both surface receptor activity and protein decreased in parallel by 50-80%. Virtually all of the lost surface receptor protein was found inside the cell, but only about 50% of all cellular Gal receptors were active. As determined by equilibrium binding studies, this decreased receptor activity reflected an overall loss of ligand binding sites with little change in binding affinity of the remaining Gal receptors for asialo orosomucoid. When ATP was restored, normal surface receptor activity and number completely recovered even in the absence of protein synthesis. We conclude that a subpopulation of Gal receptors constitutively recycles and undergoes an inactivation/reactivation cycle. In the absence of ligand, these receptors are normally internalized and then inactivated. Loss of cellular ATP blocks receptor reactivation, prevents the reappearance of receptors at the cell surface and redistributes Gal receptors as inactive receptors accumulate intracellularly. PMID- 3029057 TI - Coupling between phosphoinositide breakdown and early mitogenic events in fibroblasts. Studies with fluoroaluminate, vanadate, and pertussis toxin. AB - In the preceding paper (Paris, S., and Pouyssegur J. (1987) J. Biol. Chem. 262, 1970-1976), AlF4- and vanadate have been shown to induce inositol phosphate formation in resting hamster fibroblasts (CCL39). In this study, we show that these two phosphate analogs are good tools to explore the causal relationship between phosphoinositide breakdown and early mitogenic events. AlF4- can activate, very similarly to the mitogen alpha-thrombin: the amiloride-sensitive Na+/H+ antiport, the bumetanide-sensitive Na+/K+/Cl- co-transport, and the expression of c-myc mRNA. The link between phospholipase C activation and these early events of the mitogenic response is demonstrated by the similarity of all dose-response curves for NaF and AlCl3 and by the common sensitivity of the four events to pertussis toxin. Vanadate likewise stimulates the Na+/H+ antiport through a pertussis toxin-sensitive pathway. On longer incubations, both fluoride and vanadate were found to be toxic and failed to induce DNA synthesis. Therefore, we have used pertussis toxin to investigate the link between phospholipase C activation and commitment to DNA synthesis. We show that pertussis toxin strikingly inhibits thrombin-induced reinitiation of DNA synthesis but does not affect the stimulation by the epidermal or fibroblast growth factors, two mitogens that do not stimulate phosphoinositide breakdown in CCL39 cells. In conclusion, these studies demonstrate that activation of phospholipase C, if not an obligatory step in the action of all growth factors, plays a crucial role in the mitogenic signaling pathway of alpha-thrombin. PMID- 3029056 TI - Further evidence for a phospholipase C-coupled G protein in hamster fibroblasts. Induction of inositol phosphate formation by fluoroaluminate and vanadate and inhibition by pertussis toxin. AB - We have previously reported that alpha-thrombin induces in resting hamster fibroblasts (CCL39) the formation of inositol phosphates (IP) by activating a GTP binding protein (G protein) sensitive to pertussis toxin (Paris, S., and Pouyssegur, J. (1986) EMBO J. 5, 55-60). Here we show that IP formation in CCL39 cells can also be induced by NaF with AlCl3 and by vanadate. In the presence of Li+, IP accumulation is linear over 30 min with no detectable lag and is concentration-dependent. NaF alone is slightly stimulatory, but a marked potentiation is observed in the presence of AlCl3, by itself without effect. Maximal stimulation is obtained with 10 mM NaF and 3 microM AlCl3, and with vanadate half-maximal effect is achieved at 0.3 mM. Both stimulations are markedly inhibited (up to 80%) by pertussis toxin (half-maximal inhibition at 1-2 ng/ml). We therefore conclude that phospholipase C is stimulated by NaF plus AlCl3 (presumably acting as AlF-4) and by vanadate by direct activation of the regulatory G protein. In addition, NaF inhibits the inositol-1-phosphatase, but this effect is not potentiated by AlCl3. Similarly, vanadate inhibits inositol trisphosphate degradation. Maximal stimulations of phospholipase C by AlF-4 and vanadate are not additive, whereas they are both additive with thrombin effects. Pretreatment of cells for 15 min with the phorbol ester 12-O-tetradecanoylphorbol 13-acetate nearly completely abolishes induction of IP formation by AlF-4 and vanadate, suggesting that protein kinase C exerts a feedback negative control either on the G protein or on phospholipase C itself. An increase in cellular cyclic AMP similarly results in a marked attenuation of AlF-4-induced IP formation, indicating that activation of phospholipase C can be controlled also by cyclic AMP. However, the stimulatory effect of AlF-4 on phospholipase C is clearly dissociated from its effect on the adenylate cyclase system. PMID- 3029058 TI - Messenger RNA guanylyltransferase from Saccharomyces cerevisiae. Large scale purification, subunit functions, and subcellular localization. AB - Messenger RNA capping enzyme (GTP:mRNA guanylyltransferase) purified from yeast Saccharomyces cerevisiae consisted of two polypeptides (45 and 39 kDa) and possessed two enzymatic activities, i.e. mRNA guanylyltransferase and RNA 5' triphosphatase (Itoh, N., Mizumoto, K., and Kaziro, Y. (1984) J. Biol. Chem. 259, 13923-13929). In this paper, we describe an improved procedure suitable for the large scale purification of the enzyme. The steps include glass beads disruption of the cells and several ion-exchange and affinity column chromatographies. The enzyme was purified from kilogram quantities of yeast cells to apparent homogeneity. The purified enzyme had an approximate Mr of 180,000 and consisted of two heterosubunits of 80 and 52 kDa and had the same two enzymatic activities as above. We consider that this is the more intact form of the enzyme. Using the in situ assays on sodium dodecyl sulfate-polyacrylamide gels, RNA 5' triphosphatase, and mRNA guanylyltransferase activities were located on the 80- and 52-kDa chains, respectively. In agreement with this, the 52-kDa enzyme [32P]GMP complex was formed on incubation of the enzyme with [alpha-32P]GTP. Guinea pig antisera against purified yeast capping enzyme recognized both 80- and 52-kDa chains in Western blot analysis. The antibody did not cross-react with the enzymes from rat liver. Artemia salina, or vaccinia virus. Nuclear localization of the enzyme was demonstrated by immunofluorescence microscopy. PMID- 3029059 TI - Purification and characterization of a high molecular weight type 1 phosphoprotein phosphatase from the human erythrocyte. AB - The major Mn2+-activated phosphoprotein phosphatase of the human erythrocyte has been purified to homogeneity from the cell hemolysate. It is sensitive to both inhibitors 1 and 2 of rabbit skeletal muscle, preferentially dephosphorylates the beta subunit of the phosphorylase kinase, and dephosphorylates a broad range of substrates including phosphorylase a, p-nitro-phenyl phosphate, phosphocasein, the regulatory subunit of cyclic AMP-dependent protein kinase, and both spectrin (Km = 10 microM) and pyruvate kinase (Km = 18 microM) purified from the human erythrocyte. The purified enzyme is stimulated by Mn2+ and to a lesser extent by higher concentrations of Mg2+. The purification procedure was selected to avoid any change in molecular weight, hence subunit composition, between the crude and purified enzyme. Maintenance of the original structure is demonstrated by non denaturing gel electrophoresis and gel filtration chromatography. Gel filtration of the purified holoenzyme shows a single active component with a Stokes radius of 58 A at a molecular weight position of 180,000. Sedimentation velocity in a glycerol gradient gives a value of 6.1 for S20, w. Together these data indicate a molecular weight of about 135,000. Two bands of equal intensity appear on sodium dodecyl sulfate-gel electrophoresis at molecular weights of 61,700 and 36,300, suggesting a subunit composition of two 36,000 and one 62,000 subunits. The 36 kDa catalytic subunit can be isolated by freezing and thawing the holoenzyme or by hydrophobic chromatography of the holoenzyme. The catalytic subunit shows unchanged substrate and inhibitor specificity but altered metal ion activation. PMID- 3029060 TI - Doxorubicin enhances complement susceptibility of human melanoma cells by extracellular oxygen radical formation. AB - In two recent publications we showed that rapid inactivation of cell-bound C3b is a protective mechanism of human melanoma cells against killing by the R24 monoclonal antibody and human complement (Panneerselvam, M., Welt, S., Old, L.J., and Vogel, C.-W. (1986) J. Immunol. 136, 2534-2541) and that this protective mechanism can be inhibited by both the free and immobilized anthracycline glycoside doxorubicin (adriamycin) resulting in an enhanced complement susceptibility (Panneerselvam, M., Bredehorst, R., and Vogel, C.-W. (1986) Proc. Natl. Acad. Sci. U.S.A. 83, 9144-9148). In this paper we show that the complement enhancing effect of both free and immobilized doxorubicin is caused by the generation of reactive oxygen species including superoxide anion radical, hydrogen peroxide, and hydroxyl radical. The complement-enhancing effect of the anthracyclines can be completely inhibited by the reactive oxygen scavengers superoxide dismutase, catalase, and dimethyl sulfoxide. Consistent with this observation, 5-iminodaunorubicin, an anthracycline glycoside with an imine substituted quinone moiety and, therefore, with a significantly reduced ability to form oxygen radicals, did not cause an enhanced-complement susceptibility. The complement-enhancing effect of the anthracycline glycosides could also be inhibited by bivalent metal chelators but was unaffected by sulfhydryl-blocking reagents or glutathione. Our results suggest that the anthracycline glycosides generate in a metal- (most probably iron) dependent reaction superoxide anion radicals with subsequent formation of hydrogen peroxide and hydroxyl radicals. These reactive oxygen species then cause alterations in the melanoma cells resulting in the enhanced complement susceptibility. While the target molecule(s) of the reactive oxygen species responsible for the enhanced complement susceptibility is not known, the data obtained with immobilized doxorubicin suggest that the target molecule(s) is located in the cell membrane. PMID- 3029061 TI - Phosphorylated fructose-1,6-bisphosphatase dephosphorylating protein phosphatase from Saccharomyces cerevisiae. AB - Phosphorylation of fructose-1,6-bisphosphatase with cyclic AMP-dependent protein kinase from yeast is accompanied by a 50% decrease in the catalytic activity (Pohlig, G. and Holzer, H. (1985) J. Biol. Chem. 260, 13818-13823). Using reactivation of phoshorylated fructose-1,6-bisphosphatase as assay, a protein phosphatase was about 2,000-fold purified to electrophoretic homogeneity from Saccharomyces cerevisiae. Upon incubation with phosphorylated fructose-1,6 bisphosphatase the purified protein phosphatase not only reverses the 50% inactivation caused by phosphorylation, but also the previously observed change in the pH optimum and in the ratio of activity with Mg2+ or Mn2+. The phosphatase is strongly inhibited by heparin and fluoride. L-Carnitine, orthophosphate, pyrophosphate, and succinate inhibit to 50% at concentrations from 1 to 10 mM. The molecular mass of the native phosphatase was found to be 180,000 Da. Sodium dodecyl sulfate-gel electrophoresis suggested four subunits with a molecular mass of 45,000 Da each. Half-maximal activity was observed with 5 mM Mg2+ or Mn2+, the pH optimum of activity was found at pH 7. Using polyclonal antibodies, disappearance of 32P-labeled fructose-1,6-bisphosphatase and concomitant liberation of the expected amount of inorganic [32P] phosphate was demonstrated. PMID- 3029062 TI - Synthesis and processing of alpha-galactosidase A in human fibroblasts. Evidence for different mutations in Fabry disease. AB - The synthesis and processing of the human lysosomal enzyme alpha-galactosidase A was examined in normal and Fabry fibroblasts. In normal cells, alpha galactosidase A was synthesized as an Mr = 50,500 precursor, which contained phosphate groups in oligosaccharide chains cleavable by endoglucosaminidase H. The precursor was processed via ill-defined intermediates to a mature Mr 46,000 form. Processing was complete within 3-7 days after synthesis. In the presence of NH4Cl and in I-cell fibroblasts, the majority of newly synthesized alpha galactosidase A was secreted as an Mr = 52,000 form. For comparison, the processing and stability of alpha-galactosidase A were examined in fibroblasts from five unrelated patients with Fabry disease, which is caused by deficient alpha-galactosidase A activity. In one cell line, synthesis of immunologically cross-reacting polypeptides was not detectable. In another, the synthesis, processing, and stability of alpha-galactosidase A was indistinguishable from that in normal fibroblasts. In a third Fabry cell line, the mutation retarded the maturation of alpha-galactosidase A. Finally, in two cell lines, alpha galactosidase A polypeptides were synthesized that were rapidly degraded following delivery to lysosomes. These results clearly indicate that Fabry disease comprises a heterogeneous group of mutations affecting synthesis, processing, and stability of alpha-galactosidase A. PMID- 3029063 TI - DNA helicase II of Escherichia coli. Characterization of the single-stranded DNA dependent NTPase and helicase activities. AB - Escherichia coli helicase II has been purified to near homogeneity from cells harboring a multicopy plasmid containing the structural gene for helicase II, uvrD. In this paper a detailed description of the single-stranded DNA-dependent nucleoside 5'-triphosphatase and helicase reactions catalyzed by helicase II is presented. The results of this study suggest that nucleoside 5'-triphosphate hydrolysis provides the energy required for translocation of the enzyme along single-stranded DNA. Measurements of the rate of ATP hydrolysis using a variety of single-stranded DNAs of known structure and length suggest a processive translocation mechanism for helicase II. Single-stranded DNA coated with either Escherichia coli single-stranded DNA binding protein (SSB) or bacteriophage T4 gene 32 protein fails to support helicase II ATPase activity. Moreover, helicase II is apparently unable to displace a molecule of bound SSB protein from single stranded DNA when it is encountered in the process of translocation along a single-stranded DNA effector. The helicase reaction has been characterized using an in vitro strand displacement helicase assay. The helicase reaction requires concomitant nucleoside 5'-triphosphatase hydrolysis that is satisfied by the hydrolysis of either rATP or dATP. As the length of duplex DNA present in the partial duplex helicase substrate is increased from 71 base pairs to 343 base pairs, the fraction of duplex DNA molecules that are unwound by helicase II decreases in the absence of any accessory proteins. However, the total number of base pairs of duplex DNA unwound depends primarily on the amount of enzyme added to the helicase reaction and not on the length of the duplex DNA present in the partial duplex DNA substrate. These data suggest the number of base pairs of duplex DNA unwound is directly proportional with the concentration of helicase II in the reaction mixture. In addition, the rate of the unwinding reaction is independent of the length of the duplex DNA available for unwinding. Helicase II has been shown to dissociate from single-stranded DNA molecules infrequently acting as an ATPase. However, the enzyme dissociates from partial duplex helicase substrates more frequently. This suggests a more distributive reaction mechanism on duplex DNA than was observed on single-stranded DNA substrates. The fraction of 343-base pair partial duplex DNA molecules unwound by helicase II can be increased by the addition of appropriate concentrations of E. coli SSB to the reaction. This suggests that helicase II and SSB may act in a concerted reaction to unwind duplex DNA. PMID- 3029064 TI - Inhibition of gastric H+,K+-ATPase and acid secretion by SCH 28080, a substituted pyridyl(1,2a)imidazole. AB - A hydrophobic amine, SCH 28080, 2-methyl-8-(phenylmethoxy)imidazo(1,2a)pyridine-3 acetonitrile, previously shown to inhibit gastric acid secretion in vivo and in vitro, was also shown to inhibit basal and stimulated aminopyrine accumulation in isolated gastric glands when histamine, high K+ concentrations, or dibutyryl cAMP were used as secretagogues. Stimulated, but not basal, oxygen consumption was also inhibited. Neutralization of the acid space of the parietal cell by high concentrations of the weak base, imidazole, reduced the potency of the drug, suggesting that SCH 28080 was active when protonated. Studies on the isolated H+,K+-ATPase showed that the compound inhibited the enzyme competitively with K+, whether ATP or p-nitrophenyl phosphate were used as substrates. In contrast, the inhibition was mixed with respect to p-nitrophenyl phosphate and uncompetitive with respect to ATP. The drug reduced the steady state level of the phosphoenzyme but not the observed rate constant for phosphoenzyme formation in the absence of K+ nor the quantity of phosphoenzyme reacting with K+. The drug quenched the fluorescence of fluorescein isothiocyanate-modified enzyme and also inhibited the ATP-independent K+ exchange reaction of the H+,K+-ATPase. Its action on gastric acid secretion can be explained by inhibition of the H+,K+-ATPase by reversible complexation of the enzyme. This class of compound, therefore, acts as a reversible inhibitor of gastric acid secretion. PMID- 3029065 TI - Heterogeneity of the rat hepatic Ah receptor and evidence for transformation in vitro and in vivo. AB - The characteristics of the Ah receptor from rat liver were investigated following the incubation of cytosol with [3H]2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) under various conditions, and using DEAE- and DNA-Sepharose chromatography and sucrose density gradient centrifugation. These studies indicated that the Ah receptor can exist in three distinct forms in vitro that are dependent on the presence or absence of TCDD and the duration and temperature of incubation. The unoccupied receptor was distinguished by its elution from DEAE-Sepharose columns at 0.20-0.23 M NaCl and lack of affinity for DNA-Sepharose. Following the incubation of the unoccupied receptor with [3H]TCDD, two occupied forms were distinguished based on their overall surface charges and affinities for DNA. One of these forms was predominant following short incubations (2 h) with [3H]TCDD at a low temperature (0 degree C) and was characterized by having the same elution profile on DEAE-Sepharose as the unoccupied form, but demonstrated some affinity for DNA. Another occupied form was predominant following an incubation for a longer time (20 h, 0 degree C) or at an elevated temperature (2 h, 20 degrees C). This form had an overall surface charge that was less negative and a greater affinity for DNA. These changes in receptor characteristics were dependent on the presence of TCDD and were not accompanied by apparent changes in the sedimentation coefficients of the two occupied forms. Anion exchange chromatography of the [3H]TCDD-receptor complex extracted from hepatic nuclei of [3H]TCDD-treated rats indicated that the ligand-induced change of the unoccupied receptor to a less negatively charged form had occurred in vivo. These results indicated a biochemical heterogeneity of the Ah receptor and suggested the occurrence of a ligand- and temperature-dependent transformation process in vivo and in vitro. PMID- 3029066 TI - The metabolism of tris- and tetraphosphates of inositol by 5-phosphomonoesterase and 3-kinase enzymes. AB - Phospholipase C cleaves phosphatidylinositol 4,5-bisphosphate to form both inositol 1,4,5-trisphosphate (Ins(1,4,5)P3) and inositol 1,2-cyclic 4,5 trisphosphate (cInsP3). The further metabolism of these inositol trisphosphates is determined by two enzymes: a 3-kinase and a 5-phosphomonoesterase. The first enzyme converts Ins(1,4,5)P3 to inositol 1,3,4,5-tetrakisphosphate (InsP4), while the latter forms inositol 1,4-bisphosphate and inositol 1,2-cyclic 4-bisphosphate from Ins(1,4,5)P3 and cInsP3, respectively. The current studies show that the 3 kinase is unable to phosphorylate cInsP3. Also, the 5-phosphomonoesterase hydrolyzes InsP4 with an apparent Km of 0.5-1.0 microM to form inositol 1,3,4 trisphosphate at a maximal velocity approximately 1/30 that for Ins(1,4,5)P3. The apparent affinity of the enzyme for the three substrates is InsP4 greater than Ins(1,4,5)P3 greater than cInsP3; however, the rate at which the phosphatase hydrolyzes these substrates is Ins(1,4,5)P3 greater than cInsP3 greater than InsP4. The 5-phosphomonoesterase and 3-kinase enzymes may control the levels of inositol trisphosphates in stimulated cells. The 3-kinase has a low apparent Km for Ins(1,4,5)P3 as does the 5-phosphomonoesterase for InsP4, implying that the formation and breakdown of InsP4 may proceed when both it and its precursor are present at low levels. Ins(1,4,5)P3 is utilized by both the 3-kinase and 5 phosphomonoesterase, while cInsP3 is utilized relatively poorly only by the 5 phosphomonoesterase. These findings imply that inositol cyclic trisphosphate may be metabolized slowly after its formation in stimulated cells. PMID- 3029067 TI - Different types of hypersensitive sites in the mouse metallothionein gene region. AB - We have examined the chromatin structure of the metallothionein (MT) gene region in MT- S49 mouse lymphoma cells and in derivatives which express MT-I alone, MT II alone, or both genes. In all lines, these genes are contained in a 16-kilobase pair region between two DNase I sensitive sites: one site located 5.3 kilobase pairs 5' of MT-II (the 5' gene) is present in naked DNA and retained in the chromatin of all lines; the other site located 3.1 kilobase pairs 3' of MT-I is hypersensitive. Hypersensitivity at three other sites is dependent on the expression of MT genes. Two sites 5' of MT-II disappear, and a site 3' of MT-I appears regardless of which gene is activated. The fact that these sites respond when either gene is activated suggests that the regulation of the two genes is interdependent and that the region undergoes a general change in conformation with MT activation. In addition, a single site in the 5' region of MT-II becomes hypersensitive with activation of the gene and may be related directly to expression. PMID- 3029068 TI - Differing expression patterns and evolution of the rat kininogen gene family. AB - The present investigation using molecular cloning and sequence analysis concerns the examination of the molecular basis for different expression patterns of two types of the rat kininogen genes. We show that the low molecular weight and high molecular weight forms of K kininogens are produced from a single gene through alternative usage of two 3'-coding regions, whereas only the low molecular weight forms of T kininogens are generated as a result of several mutational changes in the high molecular weight-specifying regions of both T-I and T-II kininogen genes. The mutational changes include a nucleotide substitution at the polyadenylation/processing signal site, nucleotide deletions resulting in the frame-shift mutation, and an insertion of the type 2 Alu-equivalent sequence. Because kininogens represent a multifunctional protein comprising the proteinase inhibitory activity, the kinin moiety, and the clotting activity, these results present evidence indicating the molecular basis for the disappearance of a part of the gene functions. We also show that the K and T kininogen genes as well as the two T kininogen genes are extremely homologous, excluding and including the above mutational changes, respectively. These structural relationships allow us to envisage evolutionary processes for the generation of the rat kininogen gene family, particularly for the disappearance of a part of the gene functions. PMID- 3029069 TI - Reconstitution of rabbit thrombomodulin into phospholipid vesicles. AB - The influence of phospholipid on thrombin-thrombomodulin-catalyzed activation of protein C has been studied by incorporating thrombomodulin into vesicles by dialysis from octyl glucoside-phospholipid mixtures. Thrombomodulin was incorporated into vesicles ranging from neutral (100% phosphatidylcholine) to highly charged (30% phosphatidylserine and 70% phosphatidylcholine). Thrombomodulin is randomly oriented in vesicles of different phospholipid composition. Incorporation of thrombomodulin into phosphatidylcholine, with or without phosphatidylserine, alters the Ca2+ concentration dependence of protein C activation. Soluble thrombomodulin showed a half-maximal rate of activation at 580 microM Ca2+, whereas half-maximal rates of activation of liposome reconstituted thrombomodulin were obtained between 500 microM Ca2+ and 2 mM Ca2+, depending on the composition (protein:phospholipid) of the liposomes. The Ca2+ dependence of protein C activation fits a simple hyperbola for the soluble activator, while the Ca2+ dependence of the membrane-associated complex is distinctly sigmoidal with a Hill coefficient greater than 2.4. In contrast, the Ca2+ dependence of gamma-carboxyglutamic acid (Gla) domainless protein C activation is unchanged by membrane reconstitution (1/2 max = 53 +/- 10 microM) and fits a simple rectangular hyperbola. Incorporation of thrombomodulin into pure phosphatidylcholine vesicles reduces the Km for protein C from 7.6 +/- 2 to 0.7 +/- 0.2 microM. Increasing phosphatidylserine to 20% decreased the Km for protein C further to 0.1 +/- 0.02 microM. Membrane incorporation has no influence on the activation of protein C from which the Gla residues are removed proteolytically (Km = 6.4 +/- 0.5 microM). The Km for protein C observed on endothelial cells is more similar to the Km observed when thrombomodulin (TM) is incorporated into pure phosphatidylcholine vesicles than into negatively charged vesicles, suggesting that the protein C-binding site on endothelial cells does not involve negatively charged phospholipids. In support of this concept, we observed that prothrombin and fragment 1, which bind to negatively charged phospholipids, do not inhibit protein C activation on endothelial cells or TM incorporated into phosphatidylcholine vesicles, but do inhibit when TM is incorporated into phosphatidylcholine:phosphatidylserine vesicles. These studies suggest that neutral phospholipids lead to exposure of a site, probably on thrombomodulin, capable of recognizing the Gla domain of protein C. PMID- 3029070 TI - Proteolytic formation and properties of functional domains of thrombomodulin. AB - Thrombomodulin is an endothelial cell surface protein which complexes with thrombin to accelerate protein C activation. To gain insight into the mechanisms of thrombomodulin-membrane association, limited proteolytic digestions of thrombomodulin with trypsin and elastase were performed. Trypsin digestion resulted in two major fragments (Mr = 54,000 and 27,000), both of which bound to phosphatidylcholine/phosphatidylserine vesicles. Elastase digestion also yielded two major fragments (Mr = 50,000 and 25,000), but only the smaller fragment bound to the phospholipid vesicles. The larger fragment obtained from both enzymatic digestions retained the ability to accelerate protein C activation. The Mr = 54,000 fragment from the trypsin digest retained a high affinity for thrombin (Kd less than or equal to 0.5 nM), a Km for protein C of approximately equal to 8 microM, and a half-maximal Ca2+ dependence of 0.3 mM. The Mr = 50,000 fragment from elastase digestion had a lower affinity for thrombin (Kd approximately equal to 6 nM) than intact thrombomodulin, and the Km for protein C was decreased to 0.3 microM in the presence of 0.3 mM Ca2+. The Ca2+ dependence of protein C activation with the Mr = 50,000 fragment was distinctly different from that of thrombomodulin or the active tryptic fragment. The active elastase fragment exhibited a Ca2+ optimum at 0.3 mM and activity rapidly decreased with further increases in Ca2+. At the Ca2+ optimum, the Km for protein C was similar to that observed on endothelial cell surfaces or with thrombomodulin reconstituted into liposomes. Our data demonstrate that thrombomodulin has one or more membrane binding domains and that an active soluble form with catalytic activity can be generated by limited proteolytic digestion. Digestion with elastase appears to expose a site on thrombomodulin capable of recognizing the gamma-carboxyglutamic acid domain of protein C (residues 1-44 of the light chain). Whether this is the same site which is exposed on thrombomodulin upon incorporation into phospholipid vesicles (see accompanying manuscript) remains to be determined. PMID- 3029071 TI - A novel family of mitochondrial plasmids associated with longevity mutants of Podospora anserina. AB - We have identified a novel family of plasmids, each containing very short monomeric units, in Podospora anserina longevity mutants. These plasmids, termed small mitochondrial DNAs (sMt-DNAs), are derived from a highly ordered 368-base pair region of the mitochondrial genome. A total of five direct repeat sequences and seven significant regions of dyad symmetry (i.e. palindromes) were found within a 434-base pair mitochondrial sequence, which includes this 368-base pair region. Mitochondrial DNA rearrangements accompany the formation of these small plasmids indicating their derivation from a plastic region of the mitochondrial genome. A possible relationship between the direct repeat sequences, the palindromic regions, and the excision process is discussed. PMID- 3029072 TI - Formation of rolling-circle molecules during phi X174 complementary strand DNA replication. AB - The primosome is a mobile multiprotein priming apparatus that requires seven Escherichia coli proteins for assembly (the products of the dnaB, dnaC and dnaG genes; replication factor Y (protein n'); and proteins i, n, and n"). While the primosome is analagous to the phage T7 gene 4 protein and phage T4 gene 41/61 proteins in its DNA G-catalyzed priming function, its ability to act similarly also as a DNA helicase has remained equivocal. The role of the primosome in unwinding duplex DNA strands was investigated in the coliphage phi X174 SS(c)--- replicative form DNA replication reaction in vitro, which requires the E. coli single-stranded DNA binding protein, the primosomal proteins, and the DNA polymerase III holoenzyme. Multigenome-length, linear, double-stranded DNA molecules were generated in this reaction, presumably via a rolling circle-type mechanism. Synthesis of these products required the presence of a helicase catalyzed strand-displacement activity to permit multiple cycles of continuous complementary (-) strand synthesis. The participation of the primosome in this helicase activity was supported by demonstrating that other SS(c) DNA templates (G4 and alpha-3), which lack primosome assembly sites, failed to support significant linear multimer production and that replication of phi X174 with the general priming system (the DNA B and DNA G proteins and DNA polymerase III holoenzyme) resulted in a 13-fold lower rate of linear multimer synthesis. PMID- 3029073 TI - The molecular basis of a familial apoE deficiency. An acceptor splice site mutation in the third intron of the deficient apoE gene. AB - The molecular basis of the familial apoE deficiency was investigated by gene cloning and comparative expression studies of the normal and the deficient apoE gene. For the latter studies the apoE genes were placed under the control of the mouse metallothionein I promoter in a bovine papilloma virus vector. The studies showed that in the normal gene the mouse metallothionein I promoter directs the synthesis of normal apoE mRNA and protein. In contrast, in the deficient apoE gene the same promoter directs the synthesis of two abnormal apoE mRNA forms, which are similar to those observed in the peripheral blood monocyte macrophage cultures of the patient. Restriction analysis of the cloned gene and partial DNA sequence has shown an A to G substitution in the penultimate 3' nucleotide of the third intron of the apoE gene. This single base substitution abolishes the correct 3' splice site thus creating two abnormally spliced mRNA forms. The smaller form contains 53 nucleotides and the larger form contains the entire third intron of the apoE gene. Both of these mRNA species contain chain termination codons within the intronic sequence and code for short apoE peptides that are not detectable by gel electrophoretic techniques. These findings show that this form of familial apoE deficiency results from a point mutation in the 3' splice junction of the third intron of the apoE gene. Furthermore, the expression system we have employed to study apoE deficiency can be utilized to analyze a broad spectrum of genetic diseases associated with mRNA processing mutations. PMID- 3029074 TI - A human purine nucleoside phosphorylase deficiency caused by a single base change. AB - Purine nucleoside phosphorylase (PNP) deficiency in humans is associated with a severe defect in T-lymphocyte function. The mutant gene was cloned from one PNP deficient patient who was the offspring of a consanguineous mating. The exons and intron/exon boundaries of the mutant PNP gene were sequenced and compared with the wild-type cDNA sequence. A single base difference was found in the coding region of the mutant gene, a G to A transition in the third exon. This single base mutation alters the codon at position 89 from Glu to Lys, a result which is consistent with previously published peptide mapping data. The patient was demonstrated to be autozygous for the single base mutation on the basis of hybridization of synthetic oligomers to genomic DNA digests. A mammalian expression vector was constructed containing the entire mutant gene under the transcriptional regulation of its own promoter. In another construction, the single base mutation was reverted to the wild-type sequence by in vitro mutagenesis. An isoelectric focusing gel containing extracts of the cells transfected with the mutant and reverted PNP gene was stained histochemically for PNP activity. The proteins from a similar gel were blotted on a nitrocellulose membrane, and immunoreactive human PNP protein was visualized. Cells transfected with the mutant gene contained no human PNP activity, but expressed immunoreactive PNP which focused at an abnormally alkaline pI. Cells transfected with the reverted gene expressed human PNP activity which co-focused with human PNP from a HeLa cell control, proving that the observed single base change was responsible for the loss of catalytic function. PMID- 3029075 TI - In vitro translocation of protein across Escherichia coli membrane vesicles requires both the proton motive force and ATP. AB - The energy requirement for protein translocation across membrane was studied with inverted membrane vesicles from an Escherichia coli strain that lacks all components of F1F0-ATPase. An ompF-lpp chimeric protein was used as a model secretory protein. Translocation of the chimeric protein into membrane vesicles was totally inhibited in the presence of carbonyl cyanide m-chlorophenylhydrazone (CCCP) or valinomycin and nigericin and partially inhibited when either valinomycin or nigericin alone was added. Depletion of ATP with glucose and hexokinase resulted in the complete inhibition of the translocation process, and the inhibition was suppressed by the addition of ATP-generating systems such as phosphoenolpyruvate-pyruvate kinase or creatine phosphate-creatine kinase. These results indicate that both the proton motive force and ATP are required for the translocation process. The results further suggest that both the membrane potential and the chemical gradient of protons (delta pH), of which the proton motive force is composed, participate in the translocation process. PMID- 3029076 TI - Biosynthesis of human C-reactive protein in cultured hepatoma cells is induced by a monocyte factor(s) other than interleukin-1. AB - An in vitro cell culture system utilizing continuous human liver cells has been developed which, upon specific induction, will respond by synthesizing, de novo, the prototype acute phase reactant, C-reactive protein (CRP). Induction of CRP in vitro is not brought about by the types of hormones, steroids, and chemicals which affect other acute phase proteins. In particular, interleukin-1 thought to be directly responsible for acute phase induction is not found to be active. Direct testing of other purified biological response modifiers, i.e. alpha, beta, and gamma-interferon, interleukin-2, and tumor necrosis factor, demonstrates no inducing activity. However, we find that human peripheral blood monocytes, stimulated by endotoxin, produce a factor(s) which directly induces CRP synthesis in hepatoma cells. In addition, the human promyelocyte-like cell line HL-60 in the presence of phorbol ester and certain T-cell lines containing human retroviruses also produce this CRP-inducing factor(s). Isolation and partial purification of the CRP-inducing factor(s) indicate that it is a protein(s) with a molecular weight of approximately 30,000. PMID- 3029077 TI - Structural characterization of site-specific discontinuities associated with replication origins of minicircle DNA from Crithidia fasciculata. AB - The kinetoplast DNA of trypanosomes is comprised of thousands of DNA minicircles and 20-50 maxicircles catenated into a single network. Replication intermediates of minicircle DNA from the trypanosomatid species Crithidia fasciculata contain site-specific discontinuities in both heavy (H) and light (L) strands. These discontinuities map to two small regions situated 180 degrees apart on the minicircle; each region has two sites at which a discontinuity can occur, one on each strand. We have determined the position of these discontinuities on the minicircle DNA sequence and have characterized their structure. H-strand discontinuities occur within a 4-5-nucleotide sequence and consist of single nicks, only one of which appears to be a DNA-DNA junction. Characterization of the remaining H-strand nicks indicates a structure other than a typical DNA-DNA or DNA-RNA junction. Discontinuities on the L-strand can be either a nick or a short gap which overlaps a 12-nucleotide sequence universally conserved among minicircles from various trypanosome species. Up to 6 nucleotides are hydrolyzed from the 5' terminus facing the gap upon treatment with alkali, suggesting the presence of an RNA primer. Based on the structures of minicircle replication intermediates, we present a model for replication of minicircle DNA in which the site-specific discontinuities closely coincide with the origins of replication. PMID- 3029078 TI - Genetic demonstration of bidirectionality in the high affinity purine base transporter of mutant mouse S49 cells. AB - A mutant cell line was selected from wild type S49 lymphoblasts that expressed a novel high affinity purine base transport system not found in parental cells or any other mammalian cell line (Aronow, B., Toll, D., Patrick, J., Hollingsworth, P., McCartan, K., and Ullman, B. (1986) Mol. Cell. Biol. 6, 2957-2962). In order to determine whether this nucleobase transport system was bidirectional, mutant cell lines possessing this high affinity base transport capability were derived from a nucleoside transport-deficient derivative of an adenylosuccinate synthetase-deficient S49 cell line. The resulting progeny excreted significantly greater amounts of purine into the cell culture medium than parental cells. This purine was identified as hypoxanthine. These results demonstrate genetically that the high affinity purine base transport system can mediate both the influx and efflux of hypoxanthine. PMID- 3029079 TI - Interaction of high density lipoprotein with its receptor on cultured fibroblasts and macrophages. Evidence for reversible binding at the cell surface without internalization. AB - Cultured extrahepatic cells possess a specific high affinity receptor for high density lipoprotein (HDL) that is induced by cholesterol delivery to cells. Current results suggest that HDL receptors on cultured human fibroblasts and mouse peritoneal macrophages promote reversible binding of HDL to the cell surface without internalization of lipoprotein particles. When 125I-HDL3 was bound to cultured cells at 0 degrees C and then warmed to 37 degrees C after removal of unbound lipoprotein, most of the cell surface-bound HDL was released rapidly (t1/2 = 3 min) into the medium without entering a cellular pool that was inaccessible to digestion by trypsin at 0 degrees C. This lack of internalization of HDL was evident under conditions where internalization of 125I-low density lipoprotein and 125I-transferrin were readily detected. When cells were exposed to 125I-HDL3 at 37 degrees C, only a trace amount of iodinated apoprotein remained associated with cells after treatment of cells with trypsin. Fibroblasts treated with medium containing increasing concentrations of cholesterol exhibited a dose-dependent increase in reversible, trypsin-sensitive binding of 125I-HDL3 at 37 degrees C without an attendant increase in trypsin-resistant binding. These results suggest that reversible binding of HDL to its cell-surface receptor without subsequent endocytosis of receptor-HDL complexes is the mechanism by which HDL receptors facilitate cholesterol transport from cells. PMID- 3029080 TI - Synthetic high density lipoprotein particles. Application to studies of the apoprotein specificity for selective uptake of cholesterol esters. AB - Particles closely resembling rat high density lipoproteins (HDL) in terms of equilibrium density profile and particle size were prepared by sonication of apoA I with a microemulsion made with egg lecithin and cholesterol oleate. These particles, like authentic HDL, allowed selective uptake of their cholesterol ester moieties by cultured cells without parallel uptake of the particle itself. That uptake was saturable and competed by HDL. In rats, the plasma decay kinetics and sites of uptake of a cholesteryl ether tracer were similar whether that tracer was incorporated into synthetic or authentic HDL. Synthetic particles containing other apoproteins were made by generally the same method, but using in place of apoA-I either a mixture of rat apoCs or apoE that was either competent or reductively methylated to prevent interaction with the B/E receptor. These particles, of lower density and larger Stokes radius than those made with apoA-I, also allowed selective uptake of cholesterol esters, albeit with a lower degree of selectivity than in the case of apoA-I. Thus a specific apoprotein component in the subject lipoprotein particle is not required for selective uptake. However, selective uptake was shown to be a function of particle density or size, and part of the difference in rates of selective uptake from the particles made with various apoproteins was explained by their differences in density or size. PMID- 3029081 TI - Demonstration of two distinct high molecular weight proteases in rabbit reticulocytes, one of which degrades ubiquitin conjugates. AB - Reticulocytes contain a nonlysosomal proteolytic pathway that requires ATP and ubiquitin. By DEAE chromatography and gel filtration, we were able to fractionate the ATP-dependent system into a 30-300-kDa fraction that catalyzes the ATP dependent conjugation of ubiquitin to substrates ("Conjugation Fraction") and a high mass fraction (greater than 450 kDa) necessary for hydrolysis of the conjugated proteins. The latter contains two distinct proteases. One protease is unusually large, approximately 1500 kDa, and degrades proteins only when ATP and the conjugating fractions are added. This activity precipitates at 0-38% (NH4)2SO4 saturation and is essential for ATP-dependent proteolysis. Like crude extracts, it is labile in the absence of nucleotides and is inhibited by heparin, poly(Glu-Ala-Tyr), 3,4-dichloroisocoumarin, hemin, decavanadate, N ethylmaleimide, and various peptide chloromethyl ketones. It lacks amino peptidase and insulin-degrading activities and does not require tRNA for activity. The ubiquitin-conjugate degrading enzyme, which we suggest be named UCDEN, is inactive against substrates that cannot undergo ubiquitin conjugation. The smaller protease (670 kDa), which precipitates at 40-80% (NH4)2SO4 saturation, does not require ATP or ubiquitin and is therefore not required for ATP-dependent proteolysis. It is stimulated by N-ethylmaleimide and 3,4 dichloroisocoumarin and is stable at 37 degrees C. It hydrolyzes fluorometric tetrapeptides and proteins, including proteins which cannot be conjugated to ubiquitin. Thus, reticulocytes contain two large cytosolic proteases: one is essential for the degradation of ubiquitin conjugates, while the function of the other is uncertain. PMID- 3029082 TI - The cellular retinol binding protein II gene. Sequence analysis of the rat gene, chromosomal localization in mice and humans, and documentation of its close linkage to the cellular retinol binding protein gene. AB - Cellular retinol binding protein II (CRBP II) is an abundant, 134-residue protein present in the small intestinal epithelium. It is thought to participate in the uptake and/or intracellular metabolism of vitamin A. We have isolated and sequenced the rat CRBP II gene. Its four exons span 0.65 kilobases and are interrupted by three introns with an aggregate length of 19.5 kilobases. Southern blot hybridization analysis indicated that this gene is highly conserved in rats, mice, and humans. CRBP II belongs to a protein family that contains eight known members. Computer-assisted comparative sequence analyses indicated that a region of internal homology spans its first two exons and that oligopeptide domains specified by these first two exons exhibit significant homology to all other family members as well as to a portion of the all-trans-retinol binding domain that has previously been defined in serum retinol binding protein. The CRBP II gene was mapped in mice using recombinant inbred strains and restriction fragment length polymorphisms. It is located on chromosome 9 within 5.3 centimorgans of the phosphoglucomutase-3 locus and is closely linked (within 3.0 centimorgans) to the gene specifying a highly homologous intracellular retinol binding protein known as CRBP. Mouse-human somatic cell hybrids were used to determine that both the CRBP and CRBP II genes are located on human chromosome 3. PMID- 3029083 TI - Regulation of pantothenate kinase by coenzyme A and its thioesters. AB - Pantothenate kinase catalyzes the rate-controlling step in the coenzyme A (CoA) biosynthetic pathway, and its activity is modulated by the size of the CoA pool. The effect of nonesterified CoA (CoASH) and CoA thioesters on the activity of pantothenate kinase was examined to determine which component of the CoA pool is the most effective regulator of the enzyme from Escherichia coli. CoASH was five times more potent than acetyl-CoA or other CoA thioesters as an inhibitor of pantothenate kinase activity in vitro. Inhibition by CoA thioesters was not due to their hydrolysis to CoASH. CoASH inhibition was competitive with respect to ATP, thus providing a mechanism to coordinate CoA production with the energy state of the cell. There were considerable differences in the size and composition of the CoA pool in cells grown on different carbon sources, and a carbon source shift experiment was used to test the inhibitory effect of the different CoA species in vivo. A shift from glucose to acetate as the carbon source resulted in an increase in the CoASH:acetyl-CoA ratio from 0.7 to 4.3. The alteration in the CoA pool composition was associated with the selective inhibition of pantothenate phosphorylation, consistent with CoASH being a more potent regulator of pantothenate kinase activity in vivo. These results demonstrate that CoA biosynthesis is regulated through feedback inhibition of pantothenate kinase primarily by the concentration of CoASH and secondarily by the size of the CoA thioester pool. PMID- 3029084 TI - Circular dichroism investigation of Escherichia coli adenylate kinase. AB - We examined by circular dichroism (CD) spectroscopy in far- and near-ultraviolet three different molecular forms of Escherichia coli adenylate kinase: the wild type protein, the enzyme carboxymethylated at a single cysteine residue (Cys-77), and the thermosensitive adenylate kinase. The thermosensitive enzyme differs from the wild type protein in that a serine is substituted for a proline residue at position 87 (Gilles, A.-M., Saint Girons, I., Monnot, M., Fermandjian, S., Michelson, S., and Barzu, O. (1986) Proc. Natl. Acad. Sci. U. S. A., 83, 5798 5802). We also examined the CD spectra of isolated peptides resulting from chemical cleavage of adenylate kinase at Cys-77 (C1, residues 1-76; C2, residues 77-214). The secondary structure composition of wild type bacterial adenylate kinase (50% alpha-helix and 15% beta-sheet) was close to that derived from x-ray analysis of pig muscle enzyme (Schulz, G.E., Elzinga, M., Marx, F., and Schirmer, R. H. (1974) Nature 250, 120-123). Carboxymethylation of wild type protein did not greatly affect the CD spectrum. The secondary structure of the thermosensitive adenylate kinase was observed to be significantly different from that of the wild type enzyme (reduction in alpha-helix content to 39%). Changes in ellipticities at 222 nm as a function of temperature indicated that the melting temperature for thermosensitive adenylate kinase was 38 degrees C and that for the wild type enzyme was 54 degrees C. Isolated C1 and C2 peptides had a large proportion of unordered structures. When mixed, C1 and C2 fragments reassociated into structures resembling native, uncleaved adenylate kinase. The recovery of ordered structures, indicated by CD spectroscopy, paralleled the recovery of catalytic activity. PMID- 3029086 TI - Bacterially synthesized vertebrate calmodulin is a specific substrate for ubiquitination. AB - Calmodulin purified from bacteria which express a cloned chicken calmodulin gene can be selectively conjugated with ubiquitin, using enzymes present in reticulocyte extracts. Analyses of peptide products generated from limited proteolytic digestion of the calmodulin conjugate containing a single ubiquitin indicate that lysine 115 on calmodulin is the site of linkage. This linkage site is identical to that previously reported for calmodulin purified from Dictyostelium discoideum. Substrate-dependent ATP hydrolysis by a partially purified ubiquitin conjugation enzyme system from reticulocyte extracts was used to determine the enzyme affinity to calmodulin. Km values of 7 and 9 microM were determined for dictyostelium and the bacterially expressed calmodulin, respectively. The bacterially expressed calmodulin, unlike the Dictyostelium protein, can also form conjugates containing a 2-5 molar ratio of ubiquitin but at a slower rate than that observed for conjugation at lysine 115. Results from these studies further support our hypothesis that the post-translational methylation of lysine 115 found in most forms of calmodulin serves the important function of protecting calmodulin from ubiquitination and from degradation by the cytoplasmic ubiquitin-dependent proteolytic pathway. The capability of the bacterially expressed calmodulin to form conjugates with a high molar ratio of ubiquitin suggests that the post-translational acetylation of the N terminus of calmodulin may serve a similar function. PMID- 3029085 TI - The final step in the de novo biosynthesis of platelet-activating factor. Properties of a unique CDP-choline:1-alkyl-2-acetyl-sn-glycerol choline phosphotransferase in microsomes from the renal inner medulla of rats. AB - Final steps in the synthesis of platelet activating factor (PAF) occur via two enzymatic reactions: the acetylation of 1-alkyl-2-lyso-sn-glycero-3 phosphocholine by a specific acetyltransferase or the transfer of the phosphocholine base group from CDP-choline to 1-alkyl-2-acetyl-sn-glycerol by a dithiothreitol (DTT)-insensitive cholinephosphotransferase. Our studies demonstrate that rat kidney inner medulla microsomes synthesize PAF primarily via the DTT-insensitive cholinephosphotransferase since the specific activity of this enzyme is greater than 100-fold higher than the acetyltransferase. The two cholinephosphotransferases that catalyze the biosynthesis of phosphatidylcholine and PAF have similar Mg2+ or Mn2+ requirements and are inhibited by Ca2+. Also topographic experiments indicated that both activities are located on the cytoplasmic face of microsomal vesicles. PAF synthesis was slightly stimulated by 10 mM DTT, whereas the enzymatic synthesis of phosphatidylcholine was inhibited greater than 95% under the same conditions. The concept of two separate enzymes for PAF and phosphatidylcholine synthesis is further substantiated by the differences in the two microsomal cholinephosphotransferase activities with respect to pH optima, substrate specificities, and their sensitivities to temperature, deoxycholate, or ethanol. Study of the substrate specificities of the DTT-insensitive cholinephosphotransferase showed that the enzyme prefers a lipid substrate with 16:0 or 18:1 sn-1-alkyl chains. Short chain esters at the sn 2 position (acetate or propionate) are utilized by the DTT-insensitive cholinephosphotransferase, but analogs with acetamide or methoxy substituents at the sn-2 position are not substrates. Also, CDP-choline is the preferred water soluble substrate when compared to CDP-ethanolamine. Utilization of endogenous neutral lipids as a substrate by the DTT-insensitive cholinephosphotransferase demonstrated that sufficient levels of alkylacetylglycerols are normally present in rat kidney microsomes to permit the synthesis of physiological quantities of PAF. These data suggest the renal DTT-insensitive cholinephosphotransferase could be a potentially important enzyme in the regulation of systemic blood pressure. PMID- 3029087 TI - The mechanism of oxyperoxidase formation from ferryl peroxidase and hydrogen peroxide. AB - Formation of oxyperoxidase from the reaction of ferryl horseradish peroxidase with H2O2 is inhibited by a small amount of tetranitromethane (TNM), a powerful scavenger of superoxide anion radical. The inhibition by TNM, however, does not exceed 35% as the TNM concentration is increased above 5 microM. The stoichiometry of the reaction in the presence of TNM suggests the following equation for TNM-sensitive formation of oxyperoxidase. Ferryl peroxidase + H2O2-- -(ferric peroxidase + O2- + H+)----oxyperoxidase The kinetic study on the TNM resistant formation of oxyperoxidase suggests that the displacement of the oxygen with H2O2 takes place at the sixth coordination position at maximal rates of 0.048 and 0.054 s-1 for peroxidases A and C, respectively, at 5 degrees C. The TNM-sensitive and -resistant reactions are concluded to occur in parallel, and both yield oxyperoxidase. In either mechanism, the protonated form of ferryl peroxidase is active and the pK alpha value is 7.1 for peroxidase A and 8.6 for peroxidase C. Oxyperoxidase decomposes spontaneously with a large activation energy (23.0 kcal/mol), and the reaction of ferryl peroxidase with H2O2 reaches a steady level of oxyperoxidase, which depends on pH and the concentration of H2O2. PMID- 3029088 TI - Exon structure of a mannose-binding protein gene reflects its evolutionary relationship to the asialoglycoprotein receptor and nonfibrillar collagens. AB - Cloned cDNAs encoding mannose-binding proteins isolated from rat liver have been used to isolate one of the genes encoding this group of proteins. This gene, which encodes the minor form of binding protein (designated MBP-A), has been characterized by sequence analysis. The protein-coding portion of the mRNA for the MBP-A is encoded by four exons separated by three introns. The NH2-terminal, collagen-like portion of the protein is encoded by the first two exons. These exons resemble the exons found in the genes for nonfibrillar collagens in that the intron which divides them is inserted between the first two bases of a glycine codon and the exons do not have the 54- or 108-base pair lengths characteristic of fibrillar collagen genes. The carbohydrate-binding portion of MBP-A is encoded by the remaining two exons. This portion of the protein is homologous to the carbohydrate-recognition domain of the hepatic asialoglycoprotein receptor, which is encoded by four exons. It appears that the three COOH-terminal exons of the asialoglycoprotein receptor gene have been fused into a single exon in the MBP-A gene. The organization of the MBP-A gene is very similar to the arrangement of the gene encoding the highly homologous pulmonary surfactant apoprotein, although one of the intron positions is shifted by a single amino acid. The 3' end of a mannose-binding protein pseudogene has also been characterized. PMID- 3029089 TI - Increased cytosolic calcium. A signal for sulfonylurea-stimulated insulin release from beta cells. AB - The mechanisms by which glyburide and tolbutamide signal insulin secretion were examined using a beta cell line (Hamster insulin-secreting tumor (HIT) cells). Insulin secretion was measured in static incubations, free cytosolic Ca2+ concentration ([Ca2+]i) was monitored in quin 2-loaded cells, and cAMP quantitated by radioimmunoassay. Insulin secretory dose-response curves utilizing static incubations fit a single binding site model and established that glyburide (ED50 = 112 +/- 18 nM) is a more potent secretagogue than tolbutamide (ED50 = 15 +/- 3 microM). Basal HIT cell [Ca2+]i was 76 +/- 7 nM (mean +/- S.E., n = 141) and increased in a dose-dependent manner with both glyburide and tolbutamide with ED50 values of 525 +/- 75 nM and 67 +/- 9 microM, respectively. The less active tolbutamide metabolite, carboxytolbutamide, had no effect on [Ca2+]i or insulin secretion. Chelation of extracellular Ca2+ with 4 mM EGTA completely inhibited the sulfonylurea-induced changes in [Ca2+]i and insulin release and established that the rise in [Ca2+]i came from an extracellular Ca2+ pool. The Ca2+ channel blocker, verapamil, inhibited glyburide- or tolbutamide-stimulated insulin release and the rise in [Ca2+]i at similar concentrations with IC50 values of 3 and 2.5 microM, respectively. At all concentrations tested, the sulfonylureas did not alter HIT cell cAMP content. These findings provide direct experimental evidence that glyburide and tolbutamide allow extracellular Ca2+ to enter the beta cell through verapamil-sensitive, voltage-dependent Ca2+ channels, causing a rise in [Ca2+]i which is the second messenger that stimulates insulin release. PMID- 3029090 TI - Role of the heavy and light chains of botulinum neurotoxin in neuromuscular paralysis. AB - Botulinum neurotoxin (NT) is synthesized by Clostridium botulinum in any of seven antigenically distinct forms called types A-G. NT, when fully active, is a dichain protein, composed of two polypeptides, a heavy (H) and a light (L) chain (approximately 100,000 and approximately 50,000 Da, respectively) that are held together by noncovalent bonds and at least one disulfide bond. Two types of dichain NT, A and B, and their respective H and L chains were applied to nerve muscle (NM) preparations (phrenic nerve-hemidiaphragm of the mouse), in order to develop a broader, comparative understanding of the neuroparalytic actions of NT types. It was found that the paralysis induced by dichain NT was delayed or antagonized if NM preparations were incubated with isolated and purified H chain prior to, or during, incubation with the parent, dichain NT. NM preparations preincubated with H chain and then washed free of unbound H chain became paralyzed after subsequent incubation with L chain. Paralysis did not occur if NM preparations were incubated first with L chain, washed, and then incubated with H chain. These observations suggest that the H chain binds with specific sites on the nerve terminal. This binding appears to permit the L chain, or some combination of the L and H chain, to bring about neuroparalysis through a mechanism very similar to that of the parent, dichain NT. PMID- 3029091 TI - Activation of type I cyclic AMP-dependent protein kinases with defective cyclic AMP-binding sites. AB - Two S49 mouse lymphoma cell variants hemizygous for expression of mutant regulatory (R) subunits of type I cyclic AMP-dependent protein kinase were used to investigate functional consequences of lesions in the putative cAMP-binding sites of R subunit. Kinase activation properties of wild-type and mutant enzymes were compared using cAMP and six site-selective analogs of cAMP. Kinases from both mutant sublines were relatively resistant to cyclic nucleotide-dependent activation, but they were fully activable by at least some effectors. Relative resistances of the mutant kinases varied from about 5-fold for analogs selective for their nonmutated sites to as much as 700-fold for analogs selective for their mutated sites; resistance to cAMP was intermediate. Apparent affinities of wild type and mutant R subunits for [3H]cAMP were not appreciably different, but competition experiments with site-selective analogs of cAMP suggested that binding of cAMP to mutant R subunits was primarily to their nonmutated sites. Analyses of cooperativity in cyclic nucleotide-dependent activation of mutant kinases, synergism between site I- and site II-selective analogs in activating the mutant enzymes, and dissociation of bound cAMP from mutant R subunits provided additional evidence that the mutations in these strains selectively inactivated single classes of cAMP-binding sites: phenomena attributable in wild type enzyme to intrachain interactions between sites I and II were always absent or severely diminished in experiments with the mutant enzymes. These results confirm that R subunit sequences implicated in cAMP binding by homology with other cyclic nucleotide-binding proteins actually correspond to functional cAMP binding sites. Furthermore, occupation of either cAMP-binding site I or II is apparently sufficient for activation of cAMP-dependent protein kinase. The presence of four functional cAMP-binding sites in wild-type kinase enhances the cooperativity and sensitivity of cAMP-mediated activation. PMID- 3029092 TI - HCO3- transport in basolateral membrane vesicles isolated from rat renal cortex. AB - The mechanism of HCO3- translocation across the proximal tubule basolateral membrane was investigated by testing for Na+-HCO3- cotransport using isolated membrane vesicles purified from rat renal cortex. As indicated by 22Na+ uptake, imposing an inwardly directed HCO3- concentration gradient induced the transient concentrative accumulation of intravesicular Na+. The stimulation of basolateral membrane vesicle Na+ uptake was specifically HCO3(-)-dependent as only basolateral membrane-independent Na+ uptake was stimulated by an imposed hydroxyl gradient in the absence of HCO3-. No evidence for Na+-HCO3- cotransport was detected in brush border membrane vesicles. Charging the vesicle interior positive stimulated net intravesicular Na+ accumulation in the absence of other driving forces via a HCO3(-)-dependent pathway indicating the flow of negative charge accompanies the Na+-HCO3- cotransport event. Among the anion transport inhibitors tested, 4-4'-diisothiocyanostilbene-2,2'-disulfonic acid demonstrated the strongest inhibitor potency at 1 mM. The Na+-coupled transport inhibitor harmaline also markedly inhibited HCO3- gradient-driven Na+ influx. A role for carbonic anhydrase in the mechanism of Na+-HCO3- cotransport is suggested by the modest inhibition of HCO3- gradient driven Na+ influx caused by acetazolamide. The imposition of Cl- concentration gradients had a marked effect on HCO3- gradient-driven Na+ influx which was furosemide-sensitive and consistent with the operation of a Na+-HCO3- for Cl- exchange mechanism. The results of this study provide evidence for an electrogenic Na+-HCO3- cotransporter in basolateral but not microvillar membrane vesicles isolated from rat kidney cortex. The possible existence of an additional basolateral membrane HCO3(-)-translocating pathway mediating Na+-HCO3- for Cl- exchange is suggested. PMID- 3029093 TI - Transmembrane signaling during interleukin 1-dependent T cell activation. Interactions of signal 1- and signal 2-type mediators with the phosphoinositide dependent signal transduction mechanism. AB - The murine T lymphoma line, LBRM-33 1A5, requires synergistic signals delivered by phytohemagglutinin (PHA) and interleukin 1 (IL1) for activation to high level interleukin 2 production. The activation signals provided by PHA and IL1 were replaced by the Ca2+ ionophore, ionomycin, and the phorbol ester, 12-O tetradecanoylphorbol 13-acetate (TPA), respectively. These observations supported a two-signal model for T cell activation involving increases in intracellular Ca2+ concentration ([Ca2+]i) (signal 1) and activation of protein kinase C (signal 2) as necessary and sufficient events. However, biochemical analyses revealed that additional signals were involved in the activation of LBRM-33 cells by both receptor-dependent and -independent mediators. Both signal 1-type mediators, PHA and ionomycin, exerted pleiotropic effects at the concentrations required for synergy with signal 2-type mediators (IL1, TPA). Within 1-2 min of addition, PHA stimulated phospholipid turnover, including hydrolysis of phosphatidylinositol 4,5-bisphosphate, Ca2+ mobilization, and protein kinase C activation. The [Ca2+]i increase induced by PHA was due to influx from both intracellular and extracellular Ca2+ pools. Similarly, ionomycin increased phospholipid turnover, [Ca2+]i, and directly affected protein kinase C activity in LBRM-33 cells. In contrast, the signal 2-type mediators, TPA and IL1, appeared to act by distinct intracellular mechanisms. TPA induced a protracted association of cellular protein kinase C with the plasma membrane, consistent with the two signal activation model. Furthermore, acute TPA treatment inhibited PHA stimulated inositol phosphate release and Ca2+ mobilization, suggesting that this mediator partially antagonized signal 1 delivery. IL1, in contrast, neither activated protein kinase C directly nor did it positively modulate the coupling of signal 1-type mediators to [Ca2+]i or protein kinase C via the phosphoinositide pathway. The intracellular signal delivered by IL1 is, therefore, generated through a mechanism distinct from or distal to the activation of protein kinase C. These studies indicate that the two-signal hypothesis, in its simplest form, is inadequate to explain the signals required for the initiation of IL1-dependent T cell activation. PMID- 3029095 TI - Purification of the receptor for platelet-derived growth factor from porcine uterus. AB - The receptor for platelet-derived growth factor has been purified to homogeneity on a large scale from porcine uterus. The purification procedure utilizes solubilization of uterus membranes by Triton X-100, followed by sequential chromatographies on wheat germ agglutinin-Sepharose, fast protein liquid chromatography Mono-Q, and anti-phosphotyrosine-Sepharose. About 160 micrograms of homogeneous and functionally active 170-kDa receptor could be purified from 5 kg of uterus tissue. The pure receptor responded to platelet-derived growth factor stimulation by autophosphorylation, indicating that the receptor has a kinase domain as an integral part of the molecule. A rabbit antiserum was produced against the pure receptor, which specifically recognizes the intact 170 kDa receptor. PMID- 3029094 TI - Molecular characterization of the yeast PRT1 gene in which mutations affect translation initiation and regulation of cell proliferation. AB - Several temperature-sensitive cell division cycle (cdc) mutations differentially affect the regulatory step for cell proliferation in the yeast Saccharomyces cerevisiae. We recently found that one of these mutations, cdc63-1, resides in a gene called PRT1; other mutations in this gene had been previously shown to affect translation initiation. Here we report the molecular cloning and characterization of the PRT1 gene from yeast. Our results show that the PRT1 gene is an essential, single-copy gene which encodes a 2500-nucleotide polyadenylated transcript. The nucleotide sequence indicates that the gene could code for a protein product of Mr 88,000, which bears no overall amino acid sequence similarity to any other known protein but which contains similarity over a limited region to amino acid sequences involved in nucleotide binding. PMID- 3029097 TI - Structure determination of five sulfated oligosaccharides derived from tracheobronchial mucus glycoproteins. AB - The structure of five sulfated oligosaccharide units of highly anionic tracheobronchial mucous glycoproteins, isolated from a cystic fibrosis patient's sputum, were established. Reduced oligosaccharides (84%) were released under alkaline borohydride conditions, and the acidic oligosaccharides (63%) were isolated by Dowex 1-X2 chromatography. Following the removal of acidic oligosaccharides possessing N-acetylneuraminic acid and L-fucose by lectin affinity chromatography a heterogeneous mixture of sulfated oligosaccharides was obtained. From this fraction, five short chain monosulfated oligosaccharides (S-I to S-V) were purified by sequential separation by SynChroprep AX300 anion exchange high pressure liquid chromatography, gel filtration on Bio-Gel P-2, and high voltage paper electrophoresis. Based on the results of carbohydrate composition, sequential exoglycosidase degradation, permethylation analysis, lectin affinity chromatography, and periodate oxidation, the following structures (where GalNAcol is N-acetylgalactosaminitol) were proposed for these oligosaccharides. (formula; see text) PMID- 3029096 TI - Purification and characterization of carbon monoxide dehydrogenase, a nickel, zinc, iron-sulfur protein, from Rhodospirillum rubrum. AB - Carbon monoxide dehydrogenase (CO dehydrogenase) from Rhodospirillum rubrum was shown to be an oxygen-sensitive, nickel, iron-sulfur, and zinc-containing protein that was induced by carbon monoxide (CO). The enzyme was purified 212-fold by heat treatment, ion-exchange, and hydroxylapatite chromatography and preparative gel electrophoresis. The purified protein, active as a monomer of Mr = 61,800, existed in two forms that were comprised of identical polypeptides and differed in metal content. Form 1 comprised 90% of the final activity, had a specific activity of 1,079 mumol CO oxidized per min-1 mg-1, and contained 7 iron, 6 sulfur, 0.6 nickel, and 0.4 zinc/monomer. Form 2 had a lower specific activity (694 mumol CO min-1 mg-1) and contained 9 iron, 8 sulfur, 1.4 nickel, and 0.8 zinc/monomer. Reduction of either form by CO or dithionite resulted in identical, rhombic ESR spectra with g-values of 2.042, 1.939, and 1.888. Form 2 exhibited a 2-fold higher integrated spin concentration, supporting the conclusion that it contained an additional reducible metal center(s). Cells grown in the presence of 63NiCl2 incorporated 63Ni into CO dehydrogenase. Although nickel was clearly present in the protein, it was not ESR-active under any conditions tested. R. rubrum CO dehydrogenase was antigenically distinct from the CO dehydrogenases from Methanosarcina barkeri and Clostridium thermoaceticum. PMID- 3029098 TI - Differential expression of isoforms of the regulatory subunit of type II cAMP dependent protein kinase in rat neurons, astrocytes, and oligodendrocytes. AB - The RII-B isoform of the regulatory subunit (R) of cAMP-dependent protein kinase II is abundantly and selectively expressed in cerebral cortex (Erlichman, J., Sarkar, D., Fleischer, N., and Rubin, C. S. (1980) J. Biol. Chem. 255, 8179 8184). In contrast to the cytosolic RII-H isoform from heart and other non-neural tissues, a substantial fraction of cerebral cortex RII-B is tightly associated with cell organelles. In order to study the cellular basis for the localization and abundance of RII-B in this complex and heterogeneous tissue, rat cerebral cortex was fractionated into highly purified populations of neurons, astrocytes, and oligodendrocytes. In neurons and astrocytes more than 80% of the total cAMP binding activity is contributed by RII subunits, whereas the myelin-producing oligodendrocytes contain nearly equal proportions of RI (from protein kinase I) and RII. Approximately 70% of RII and RI subunits are associated with the particulate fraction in each of the three types of brain cells. The nature of the RII isoforms expressed in the cytosolic and particulate fractions of the purified brain cells was established by performing Western immunoblot and indirect immunoprecipitation analyses with selective and sensitive polyclonal antibodies directed against RII-B. Astrocytes and neurons exhibit high levels of RII-B, whereas oligodendrocytes contain the RII-H isoform. Thus, the expression of RII isoforms is not uniform among brain cells that are anatomically and developmentally related. Rather, it appears that RII-B and RII-H are expressed in a cell-specific fashion within cerebral cortex and this might reflect an RII mediated adaptation of protein kinase II to the specialized metabolic and functional roles of neurons, astrocytes, and oligodendrocytes. PMID- 3029099 TI - Monovalent cation dependence of tissue plasminogen activator synthesis by HeLa cells. AB - HeLa cells synthesize and secrete increased levels of tissue plasminogen activator (tPA) when incubated for 18 h with 10-20 nM phorbol myristate acetate. This response was inhibited by a number of conditions which affect intracellular Na+ and K+ concentrations. Removing extracellular Na+, while maintaining isotonicity with choline+, reduced the secretion of both functional and antigenic tPA in a linear fashion. A series of cardiac glycosides and related compounds strongly inhibited tPA secretion with the following rank order of potency: digitoxin = ouabain greater than digoxin greater than digitoxigenin greater than digoxigenin greater than digitoxose greater than digitonin. These compounds also inhibited cellular Na+/K+-ATPase activity over an identical concentration range. Two compounds which selectively increase cellular permeability to K+, valinomycin, and nigericin, strongly inhibited tPA secretion, with IC50 values of approximately 50 nM. In contrast, monensin, which selectively increases cellular permeability to Na+, was much less active. Valinomycin, but not nigericin, also inhibited cellular Na+/K+-ATPase activity. Phorbol myristate acetate, 5-20 nM, increased Na+/K+-ATPase activity up to 2-fold and tPA secretion up to 15-fold. We conclude that the secretion of tPA by HeLa cells treated with phorbol myristate acetate proceeds via a mechanism which requires extracellular Na+ and a functional Na+/K+-ATPase ("sodium pump") enzyme. PMID- 3029100 TI - Regulation of adrenergic receptor function by phosphorylation. I. Agonist promoted desensitization and phosphorylation of alpha 1-adrenergic receptors coupled to inositol phospholipid metabolism in DDT1 MF-2 smooth muscle cells. AB - Continuous exposure of DDT1 MF-2 smooth muscle cells to 10-100 microM norepinephrine results in a dramatic attenuation of the ability of norepinephrine to stimulate inositol phospholipid hydrolysis via alpha 1-adrenergic receptors (alpha 1-AR). In addition to the functional desensitization, norepinephrine exposure also reduces the number of accessible cell surface alpha 1-AR as assayed by [3H]prazosin binding at 4 degrees C. Desensitization of the cells with norepinephrine results in an increase in the phosphorylation of the Mr 80,000 alpha 1-AR ligand binding peptide (2.4 +/- 0.2 mol of 32P per mol of alpha 1-AR; n = 5) when compared to control cells (1.1 +/- 0.1 mol of 32P per mol of alpha 1 AR; n = 5). The time courses of these three processes are all comparable being half-maximal within 1-2 min. These norepinephrine-promoted effects can be prevented by the alpha 1-AR receptor antagonist phentolamine indicating that they are mediated via the alpha 1-AR. Treatment of cells with the vasoactive peptide bradykinin (10 microM) induces desensitization of alpha 1-AR function similar to that induced by tumor-promoting phorbol ester treatment (Leeb-Lundberg, L. M. F., Cotecchia, S., Lomasney, J. W., DeBernardis, J. F., Lefkowitz, R. J., and Caron, M. G. (1985) Proc. Natl. Acad. Sci. USA 82, 5651-5655). Both treatments also result in phosphorylation of the alpha 1-AR, with stoichiometries of 1.7 +/- 0.1 (bradykinin; n = 5) and 3.6 +/- 0.1 (PMA; n = 5) mol of 32P/mol of alpha 1-AR. However, neither phorbol esters nor bradykinin reduce the number of accessible cell surface alpha 1-AR. Similar phosphopeptide maps are obtained from tryptic phosphopeptides generated from phosphorylated alpha 1-AR derived from cells treated with norepinephrine, phorbol 12-myristate 13-acetate, and bradykinin. Phosphoamino acid analysis reveals that the various agents induce phosphorylation on both serine and threonine residues. Thus, phosphorylation of receptors linked to the inositol phospholipid/Ca2+ signaling pathway may represent an important mechanism of regulation of receptor responsiveness. PMID- 3029102 TI - Structure of the copper sites in membrane-bound cytochrome c oxidase. AB - The structures of membrane proteins are difficult to obtain by crystallography and may be altered by the detergents used in their extraction. X-ray absorption spectroscopy has been used to identify the structures of the copper atoms of the membrane-bound enzyme in mitochondria and in submitochondrial particles at respective concentrations of 100 and 200 micron of molar copper. To within the experimental error, the x-ray absorption spectra of the copper atoms of the membrane-bound and the Yonetani (Yonetani, T. (1961) J. Biol. Chem. 236, 1680 1688) purified oxidase are identical; all detectable shells of the active site indicate no alteration of structural parameters. Significant differences are found when compared to the Hartzell-Beinert (Hartzell, R. C., and Beinert, H. (1974) Biochim. Biophys. Acta 368, 318-338) preparation. Extended x-ray absorption fine structure technology is now adequate for the direct studies of membrane proteins in situ in their natural environment. PMID- 3029101 TI - Regulation of adrenergic receptor function by phosphorylation. II. Effects of agonist occupancy on phosphorylation of alpha 1- and beta 2-adrenergic receptors by protein kinase C and the cyclic AMP-dependent protein kinase. AB - The present study was undertaken to determine the ability of protein kinase C and protein kinase A to directly phosphorylate the purified alpha 1- and beta 2 adrenergic receptors (AR). Both the catalytic subunit of protein kinase A and the protein kinase C, purified from bovine heart and pig brain, respectively, are able to phosphorylate the purified alpha 1-AR from DDT1 MF-2 smooth muscle cells. Occupancy of the receptor by an alpha 1 agonist, norepinephrine (100 microM), increases the rate of phosphorylation by protein kinase C but not by protein kinase A. The maximum stoichiometry of phosphorylation obtained is not affected by the agonist and reached 3 mol of PO4/mol of receptor for protein kinase C and 1 mol of PO4/mol of receptor for protein kinase A. The phosphopeptide maps of the trypsinized alpha 1-AR phosphorylated by each kinase differ drastically. The beta 2-AR purified from hamster lungs can also be phosphorylated by the two kinases. In contrast to the alpha 1-AR, the occupancy of the beta 2-AR by the agonist isoproterenol (20 microM) increases the rate of phosphorylation of the beta 2-AR by protein kinase A but not by protein kinase C. The maximum amount of phosphate incorporated into the receptor is not affected in either case by the agonist and reaches 1 mol of PO4/mol of receptor with protein kinase A and 0.4 mol of PO4/mol of receptor with protein kinase C. The phosphopeptide maps of the trypsinized receptor phosphorylated by either kinase reveal similar profiles. Thus, both alpha 1-AR and beta 2-AR are substrates for protein kinase A and protein kinase C. Agonist occupancy of the two receptors facilitates their phosphorylation only by the protein kinase coupled to their own signal transduction pathway. These observations suggest that "feedback" and "cross-system" phosphorylation may represent distinct and differently regulated mechanisms of modulation of receptor function. PMID- 3029103 TI - The interrelationship between cAMP-dependent alpha and beta subunit phosphorylation in the regulation of phosphorylase kinase activity. Studies using subunit specific phosphatases. AB - This study addresses the function of multisite phosphorylation of phosphorylase kinase catalyzed by the cAMP-dependent protein kinase. Using subunit specific protein phosphatases (the polycation-stimulated and ATP-, Mg2+-dependent enzymes), we show that the degree of phosphorylation of both the alpha and beta subunits modulates phosphorylase kinase activity. beta subunit phosphorylation is essential for activation and, independent of the degree of alpha subunit phosphorylation, enzyme fully dephosphorylated in the beta subunit is completely inactivated. alpha Subunit phosphorylation does, however, also regulate activity, and enzyme fully or partially phosphorylated in the beta subunit is inactivated as a consequence of alpha subunit dephosphorylation. The extent of inactivation caused by alpha subunit dephosphorylation is linearly dependent on the phosphorylation state of the beta subunit. Three peptide sites on the alpha subunit are phosphorylated by the cAMP-dependent protein kinase; the site primarily affecting activity is the one that is initially phosphorylated. These data provide evidence that subunit interrelationships play an important role in the regulation of phosphorylase kinase by multisite phosphorylation. PMID- 3029104 TI - Subunit phosphorylation and activation of phosphorylase kinase in perfused rat hearts. AB - The potential correlations between phosphorylase kinase subunit phosphorylation and activation have been examined using 32P-perfused rat hearts exposed to a variety of hormonal stimuli. Phosphate incorporation was measured after isolation of the enzyme by immunoprecipitation from heart extracts. Time courses of catecholamine or glucagon treatment produced a rapid rise in both the activity and the beta subunit phosphorylation of the enzyme, and a slightly slower increase in alpha' subunit phosphorylation. For short durations of catecholamine stimulation, the ratio of phosphate in the alpha' versus beta subunit was dependent upon hormone dose. After removal of hormone, both inactivation and alpha' subunit dephosphorylation were fairly slow, while the beta subunit was dephosphorylated more rapidly. For all of the above conditions, activation correlated with both alpha' and beta subunit phosphorylation. The maximum level of phosphate incorporation observed in response to hormonal stimulation is estimated to be approximately 1.3-1.7 mol of [32P]phosphate/mol of (alpha' beta gamma delta)4, divided about equally between the alpha' and beta subunits. When hearts were treated with hormone either in the absence of added calcium or in the presence of a calcium channel blocker, the time courses of subunit phosphorylation and activation were similar to those seen with standard perfusion conditions, suggesting that if any Ca2+-dependent autophosphorylation of phosphorylase kinase were occurring it does not make a major contribution to the observed hormonal responses. The complicated relationships observed here between phosphorylase kinase subunit phosphorylation and activation for the most part provide physiological affirmation of the patterns observed in vitro, but they also show some possible differences of potential interest. PMID- 3029105 TI - Characterization of the isolated rat flexor digitorum brevis for the study of skeletal muscle phosphorylase kinase phosphorylation. AB - The flexor digitorum brevis skeletal muscle, a nearly homogeneous fast-twitch oxidative glycolytic fiber type, has been examined for its suitability to explore the regulation of phosphorylase kinase by multisite phosphorylation. A characterization of the adrenergic response of glycogenolytic enzymes, together with the previous data on contractile properties (Carlsen, R. C., Larson, D. B., and Walsh, D. A. (1985) Can. J. Physiol. Pharm. 63, 958-965), has demonstrated that this muscle is stably maintained for the several hours necessary for phosphorylation studies. The phosphorylase kinase in this muscle is primarily the alpha' isozyme, suggesting that the alpha versus alpha' isozyme distribution in muscle is related more to oxidative capacity than to fiber contractile characteristics. Using this muscle system, beta-adrenergic activation of phosphorylase kinase was observed to occur with concomitant phosphorylation of both the alpha' and beta subunits, with the total in the alpha' subunit being approximately 3-fold greater. Similarly, deactivation, following initial adrenergic activation, occurred concomitantly with the dephosphorylation of the two subunits. These results are compatible with the conclusions drawn from previous studies of the isolated enzyme and of the enzyme in perfused rat cardiac muscle, that both alpha' (or alpha) and beta subunit phosphorylation regulate phosphorylase kinase activity. PMID- 3029106 TI - Alteration of receptor/G-protein interaction by putative endogenous protein kinase activity in Dictyostelium discoideum membranes. AB - Membranes of Dictyostelium discoideum cells were incubated under phosphorylation conditions and washed, and the effects on cAMP binding to chemotactic receptors in the absence and presence of guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S) were investigated. Most experiments were done with adenosine 5'-O-(3 thiotriphosphate) (ATP gamma S), which is a good substrate for many kinases, but the product, protein phosphorothioate, is not easily hydrolyzed by phosphatases. Pretreatment of membranes under phosphorylating conditions with MgATP gamma S alters the site heterogeneity of the cAMP-binding forms, without a significant effect on the total number of binding sites. A similar effect was induced by GTP gamma S under nonphosphorylation conditions. The effects of MgATP gamma S were rapid (t1/2 = 1 min), irreversible, and not induced by Mg2+ or ATP gamma S alone or by magnesium adenylyl imidodiphosphate and magnesium adenylyl (beta, gamma methylene)diphosphate. MgATP induced a smaller inhibition than MgATP gamma S, which was potentiated by the addition of exogenous cAMP-dependent protein kinase. The effect of MgATP was rapidly reversible; reversibility was reduced by the phosphatase inhibitor NaF. These results suggest that the effects of MgATP gamma S are mediated by an endogenous protein kinase. The major 35S-thiophosphorylated band detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis was a protein with Mr = 36,000. The phosphorylation of a protein with the molecular weight of the cAMP receptor (Mr = 40,000-45,000) was not observed. PMID- 3029107 TI - cDNA cloning and amino acid sequence of bovine brain 2',3'-cyclic-nucleotide 3' phosphodiesterase. AB - 2',3'-Cyclic-nucleotide 3'-phosphodiesterase (EC 3.1.4.37) has been widely used as a marker for myelin-oligodendrocytes in the central nervous system. Evidence has been provided that the enzyme is identical with one of the Wolfgram proteins of central nervous system myelin. The amino acid sequence of bovine 2',3'-cyclic nucleotide 3'-phosphodiesterase was determined by both protein and cDNA sequence analyses. Protein sequence analysis was done on bovine elastase 2',3'-cyclic nucleotide 3'-phosphodiesterase, a low molecular weight enzyme obtained by solubilization with pancreatic elastase (EC 3.4.21.36) (Nishizawa, Y., Kurihara, T., and Takahashi, Y. (1980) Biochem. J. 191, 71-82; Kurihara, T., Nishizawa, Y., Takahashi, Y., and Odani, S. (1981) Biochem. J. 195, 153-157). Based on the carboxyl-terminal sequence of bovine elastase 2',3'-cyclic-nucleotide 3' phosphodiesterase, synthetic oligodeoxyribonucleotides were prepared and used as probes for screening a cDNA library of bovine brain. A cDNA of 2305 base pairs was obtained and sequenced, and the complete amino acid sequence of bovine 2',3' cyclic-nucleotide 3'-phosphodiesterase was deduced. Bovine 2',3'-cyclic nucleotide 3'-phosphodiesterase deduced contains 400 amino acids including initiation methionine and has a molecular weight of 44,850. Bovine elastase 2',3' cyclic-nucleotide 3'-phosphodiesterase corresponds to the 236 amino acids of bovine 2',3'-cyclic-nucleotide 3'-phosphodiesterase. RNA blot analysis revealed a single-species mRNA of about 2600 bases. PMID- 3029108 TI - Site-specific mutagenesis of the alpha-helices of calmodulin. Effects of altering a charge cluster in the helix that links the two halves of calmodulin. AB - Alteration of residues 82-84 in the alpha-helix that links the two halves of calmodulin results in a differential effect on activator activity. Previous studies (Lukas, T. J., Burgess, W. H., Prendergast, F. G., Lau, W., and Watterson, D. M. (1986) Biochemistry 25, 1458-1464) indicated the importance of positive charge clusters in the calmodulin-binding protein, myosin light chain kinase. This suggested the possible importance of complementary negative charge clusters in calmodulin. By using an efficient cassette mutagenesis approach and a synthetic calmodulin gene (Roberts, D. M., Crea, R., Malecha, M., Alvarado Urbina, G., Chiarello, R. H., and Watterson, D. M. (1985) Biochemistry 24, 5090 5098), this possibility was directly addressed by engineering a new calmodulin, VU-8 calmodulin, in which the glutamate cluster at residues 82-84 in the synthetic gene product (VU-1 calmodulin) was replaced by three lysines. VU-8 calmodulin activated phosphodiesterase to the same maximal extent as VU-1 calmodulin, although there was an alteration in the concentration of calmodulin required for half-maximal stimulation. In contrast, myosin light chain kinase was activated to only 30% of maximal activity and NAD kinase was not activated. These results provide insight into the functional role of the unusual central helix structure found in the calmodulin family of proteins and indicate that different, although possibly overlapping, chemical complementarities are employed in the interaction between calmodulin and its various physiological targets. PMID- 3029109 TI - Factors involved in specific transcription by mammalian RNA polymerase II. Purification and functional analysis of initiation factors IIB and IIE. AB - Two general transcription factors (IIE and IIB) (TF) were purified from HeLa cell nuclear extracts and shown to be absolutely required, along with two additional factors (IIA and IID) and RNA polymerase II, for specific transcription initiation at the adenovirus major late promoter. TFIIB and TFIIE were also required, in addition to TFIIA, TFIID, RNA polymerase II, and the adenovirus 2 major late promoter, for the formation of a (preinitiation) complex that could initiate transcription (upon addition of nucleoside triphosphates) in the presence of heparin concentrations which inhibited the action of unbound factors. Glycerol gradient analyses indicated independent interactions of TFIIE with TFIIB and with the purified RNA polymerase II, but not with RNA polymerase III. Transcription factors IIB and IIE were also shown to be required for specific initiation of transcription from several cellular and viral class II promoters. PMID- 3029110 TI - Isolation of collagen binding proteins from embryonic chicken corneal epithelial cells. AB - In the present paper, we report the isolation and characterization of embryonic corneal membrane glycoproteins that demonstrate specific affinity for collagen. Two collagen binding proteins have been isolated: a novel 70-kDa protein and a 47 kDa protein which is apparently similar to that reported by Kurkinen et al. (Kurkinen, M., Taylor, A., Garrels, J. I., and Hogan, B. (1984) J. Biol. Chem. 259, 5915-5922). Both proteins label metabolically with [35S]methionine and [3H] glucosamine. 125I iodination of cell surface proteins revealed that the two collagen binding proteins are expressed on the epithelial cell surface. The 70 kDa protein appears to be an integral membrane protein, whereas the 47-kDa protein can be removed from membranes by alkali treatment. The isolated proteins exhibit binding to native type IV collagen as well as heat-denatured type I collagen. It seems likely that we have isolated, at least in part, the cell surface receptor or receptor complex that binds collagen to the basal surface of epithelia. PMID- 3029111 TI - Assembly of the mitochondrial membrane system. MRP1 and MRP2, two yeast nuclear genes coding for mitochondrial ribosomal proteins. AB - Nuclear respiratory deficient mutants of Saccharomyces cerevisiae impaired in mitochondrial protein synthesis have been screened for lesions in ribosomal protein constituents. Two mutants, each representative of a separate pet complementation group, have been analyzed. One of the mutants, E795, was found to have altered mitochondrial ribosomes as evidenced by the absence of some ribosomal proteins. The second mutant studied, C167, appeared to have more grossly altered ribosomes that could not be isolated by standard preparative procedures. In addition to being defective in mitochondrial protein synthesis, the mutants exhibit an absence of "a" and "b" type cytochromes, are partially blocked in processing of intron bI4 of the apocytochrome b gene, have reduced levels of mitochondrial 15 S rRNA, and convert to rho- and rho 0 mutants at a high frequency. The wild type genes MRP1 and MRP2 were cloned by transformation of the pet mutations in E795 and C167, respectively, with a recombinant plasmid library of wild type yeast genomic DNA. MRP1 codes for a basic protein of 37 kDa with no significant homology to any known prokaryotic or eukaryotic ribosomal protein. MRP2 codes for a 14-kDa polypeptide homologous to protein S14 of the Escherichia coli small ribosomal subunit and to a chloroplast-encoded component of chloroplast ribosomes. The levels of MRP1 and MRP2 mRNAs were examined in glucose-repressed cells and in cells undergoing adaptation to aerobic metabolism of ethanol. The steady state concentrations of the mRNAs increased during the first 3 h of derepression, indicating that expression of these mitochondrial ribosomal protein genes is transcriptionally regulated by glucose in a fashion analogous to respiratory carriers such as cytochrome c. PMID- 3029112 TI - Inhibition of methylation at two internal N6-methyladenosine sites caused by GAC to GAU mutations. AB - We previously have mapped N6-methyladenosine (m6A) sites within the genomic RNA of Rous sarcoma virus (RSV). The results of that study and of experiments using inhibitors of methylation suggest that m6A might be involved in mRNA processing events. We describe an approach for directly analyzing the function of m6A in RNA and for studying the sequence specificity of the m6A methylase. Two sites of methylation in RSV (nucleotides 7414 and 7424) were altered by oligonucleotide directed mutagenesis. The highly conserved GAC consensus sequence at those sites was changed to GAU. The new sequences were no longer methylated in the RSV genomic RNA; the GAC sequence was required for efficient base modification at those two adenosines. The altered m6A pattern did not affect viral RNA processing or the viral life cycle within infected cells. PMID- 3029113 TI - Neuropeptide Y inhibits cardiac adenylate cyclase through a pertussis toxin sensitive G protein. AB - Neuropeptide Y, a major neuropeptide and potent vasoconstrictor, inhibited isoproterenol-stimulated adenylate cyclase activity in cultured rat atrial cells as well as in atrial membranes. Prior treatment of the cells with pertussis toxin blocked the inhibitory action of neuropeptide Y. Pertussis toxin is known to uncouple the receptors for other inhibitors of adenylate cyclase by ADP ribosylation of the alpha-subunit of Gi, the inhibitory guanine nucleotide binding component of adenylate cyclase. The toxin specifically catalyzed the ADP ribosylation of a 41-kilodalton atrial membrane protein which corresponded to the Gi subunit. These results suggest that neuropeptide Y may mediate some of its physiological effects through specific receptors linked to the inhibitory pathway of adenylate cyclase. PMID- 3029114 TI - The receptor for 2,3,7,8-tetrachlorodibenzo-p-dioxin in the mouse hepatoma cell line Hepa 1c1c7. A comparison with the glucocorticoid receptor and the mouse and rat hepatic dioxin receptors. AB - The molecular properties of the receptor for 2,3,7,8-tetrachlorodibenzo-p-dioxin in the mouse hepatoma cell line Hepa 1c1c7 were investigated. The receptor was found to represent a highly asymmetrical molecule with a sedimentation coefficient, s20,w, of approximately 8 S, a Stokes radius of 7-8 nm, and a calculated Mr approximately equal to 260,000-300,000. In comparison, the Hepa 1c1c7 glucocorticoid receptor in analogy to the glucocorticoid receptor in general as well as the C57BL/6 mouse and rat hepatic dioxin receptors are molecules with an s20,w value of 4-5 S, a Stokes radius of approximately 6 nm, and a calculated Mr approximately equal to 100,000. In the presence of 20 mM sodium molybdate, a large Mr approximately equal to 270,000-310,000 form of the Hepa 1c1c7 glucocorticoid receptor is stabilized which is hydrodynamically indistinguishable from the Mr approximately equal to 260,000-300,000 Hepa 1c1c7 dioxin receptor. Sodium molybdate does not have any effect on the molecular properties of the Hepa 1c1c7 dioxin receptor. In conclusion, the large form of dioxin receptor present in Hepa 1c1c7 mouse hepatoma cells in the absence of sodium molybdate is strikingly similar to molybdate-stabilized steroid hormone receptors as well as the molybdate-stabilized form of the dioxin receptor previously demonstrated in rat hepatic cytosol. Therefore, the Hepa 1c1c7 dioxin receptor might offer an interesting model for studies on the structure and function of Mr approximately equal to 300,000 forms of soluble receptors. PMID- 3029115 TI - Human plasma cholesteryl ester transfer protein enhances the transfer of cholesteryl ester from high density lipoproteins into cultured HepG2 cells. AB - The role of human plasma cholesteryl ester transfer protein (CETP) in the cellular uptake of high density lipoprotein (HDL) cholesteryl ester (CE) was studied in a liver tumor cell line (HepG2). When HepG2 cells were incubated with [3H]cholesteryl ester-labeled HDL3 in the presence of increasing concentrations of CETP there was a progressive increase in cell-associated radioactivity to levels that were 2.8 times control. The CETP-dependent uptake of HDL-CE was found to be saturated by increasing concentrations of both CETP and HDL. The CETP dependent uptake of CE radioactivity increased continuously during an 18-h incubation. In contrast to the effect on cholesteryl ester, CETP failed to enhance HDL protein cell association or degradation. Enhanced uptake of HDL cholesteryl ester was shown for the d greater than 1.21 g/ml fraction of human plasma, partially purified CETP, and CETP purified to homogeneity, but not for the d greater than 1.21 g/ml fraction of rat plasma which lacks cholesteryl ester transfer activity. HDL cholesteryl ester entering the cell under the influence of CETP was largely degraded to free cholesterol by a process inhibitable by chloroquine. CETP enhanced uptake of HDL [3H]CE in cultured smooth muscle cells and to a lesser extent in fibroblasts but did not significantly influence uptake in endothelial cells or J774 macrophages. These experiments show that, in addition to its known role in enhancing the exchange of CE between lipoproteins, plasma CETP can facilitate the in vitro selective transfer of CE from HDL into certain cells. PMID- 3029116 TI - Chemical synthesis and expression of a cassette adapted ubiquitin gene. AB - A gene encoding the yeast ubiquitin was chemically synthesized and expressed in yeast under regulatory control of the copper metallothionein (CUP1) promoter. The gene was assembled in a one-step ligation reaction from eight oligonucleotide fragments ranging in length from 50 to 64 nucleotides. To facilitate mutagenesis and gene fusion studies, eight unique 6-base-cutting restriction enzyme sites were placed in the reading frame which did not alter the encoded protein sequence or force the utilization of rare codons. In a copper-resistant yeast strain (CUP1r), expression of the gene was induced by copper to approximately 5% of the total yeast proteins, as determined by Coomassie-stained polyacrylamide gels. The protein, purified from yeast, reacted with ubiquitin-specific antibodies and was found to be biologically active in supporting ubiquitin-dependent protein degradation in vitro. PMID- 3029117 TI - Studies on the interactions between the cyclic nucleotide-binding sites of cGMP dependent protein kinase. AB - The rate and equilibrium kinetics of [3H]cGMP binding to the two rapidly exchanging and two slowly exchanging sites of dimeric cGMP-dependent protein kinase from bovine lung were studied. As observed by McCune and Gill (McCune, R. W., and Gill, G. N. (1979) J. Biol. Chem. 254, 5083-5091), unlabeled cGMP retarded the dissociation of [3H]cGMP bound to the "slow" site. This effect was due to interaction of unlabeled cGMP with the "rapid" rather than the slow site. First, the potencies of unlabeled cGMP and a number of cGMP analogs correlated nearly perfectly with their affinities for the rapid site. Second, the rate of dissociation in the absence of unlabeled ligand was independent of the degree of saturation of the slow sites. Third, unlabeled ligand inhibited the rate of dissociation more (about 10-fold) than theoretically predicted (maximum 2-fold) from interaction between two similar sites in one macromolecule. A favorable free energy coupling appeared to exist between the rapid and slow sites but not between the slow sites. cGMP associated faster to the slow site than the rapid site. Mg/ATP decreased the rate of association to either site by 50% and increased about ten-fold the rate of dissociation from the slow site. The dissociation of cGMP from the slow site could be described by a single activation energy (Ea = 71 kJ X mol-1) for the whole temperature range (0-37 degrees C) tested. These data indicated that the cyclic nucleotide-binding sites of the cGMP kinase are kinetically more homologous to those in the cAMP-dependent protein kinases than previously recognized. PMID- 3029118 TI - Estimation of the number and turnover rate of Na+/H+ exchangers in lymphocytes. Effect of phorbol ester and osmotic shrinking. AB - Rat thymic lymphocytes possess an amiloride-sensitive Na+/H+ exchanger in their plasma membrane. Kinetic studies revealed that 5-(N-methyl-N-isobutyl)amiloride (MIA) was a more potent inhibitor of the antiport than amiloride (cf. apparent Ki of 174 nM and 6 microM, respectively). Inhibition by MIA was rapid (less than 5 s) and readily reversible. [3H]MIA binding to whole cells was assayed by rapid centrifugation following short (5 s) incubations to minimize nonspecific binding. A saturable binding component (Kd approximately equal to 170 nM) which was displaced by amiloride was detected. In contrast, there was no significant amiloride-displaceable binding to human erythrocytes, which have comparatively little Na+/H+ exchange activity. In lymphocytes, the ability of amiloride and several of its analogs to displace [3H]MIA correlated with their potency as inhibitors of the antiport. Both kinetic and binding studies revealed that extracellular H+, but not Na+, inhibited the interaction of MIA with its receptor(s). Taken together, these data suggest that [3H]MIA binds to the Na+/H+ exchanger. Scatchard analysis revealed that [3H]MIA bound to a maximum of 8000 high affinity sites/cell. Activation of Na+/H+ exchange by osmotic shrinking or by the phorbol ester 12-O-tetradecanoylphorbol 13-acetate was not accompanied by a significant change in [3H]MIA binding. Given an upper limit of 8000 functional sites/thymocyte, we estimate that the turnover number of each maximally activated exchanger is at least 2000 cycles/s. PMID- 3029119 TI - Recombinant human interleukin 1-stimulated Na+/H+ exchange is not required for differentiation in pre-B lymphocyte cell line, 70Z/3. AB - Interleukin-1 a polypeptide hormone produced by activated macrophages is a mixture of at least two proteins, interleukin-1 alpha (IL-1 alpha) and interleukin-1 beta (IL-1 beta). We have previously shown that macrophage-derived interleukin-1 induced new kappa light chain synthesis for surface IgM expression in a murine pre-B like cell line 70Z/3, a finding associated with an early amiloride-sensitive rise in the total intracellular sodium concentration. Because IL-1 alpha and IL-1 beta are structurally quite different, in this study their effect on 70Z/3 was examined separately. The results show that both human rIL-1 alpha and rIL-1 beta induce the differentiation of 70Z/3, but a higher concentration of rIL-1 beta compared to rIL-1 alpha is needed for a maximal response. At saturating concentrations, both rIL-1 alpha and rIL-1 beta induce a simultaneous rise in intracellular pH and sodium concentration. Because rIL-1 mediated intracellular alkalinization and sodium rise are amiloride sensitive, they likely occur through stimulation of the Na+/H+ exchanger across the cell membrane. Inhibition of the Na+/H+ antiport with an amiloride analog did not have an effect on rIL-1 induced surface IgM expression or the rIL-1-mediated increase in kappa light chain specific mRNA level. Therefore, these results indicate that an increase in pHi or [Na]i is not required for IL-1 induced 70Z/3 differentiation. PMID- 3029120 TI - Isolation and characterization of the yeast gene coding for the alpha subunit of mitochondrial phenylalanyl-tRNA synthetase. AB - The respiratory defect of pet mutants of Saccharomyces cerevisiae assigned to complementation group G120 has been ascribed to their inability to acylate the mitochondrial phenylalanyl tRNA. A fragment of wild type yeast genomic DNA capable of complementing the genetic lesion of G120 mutants has been cloned by transformation with a yeast genomic recombinant library of a representative mutant from this complementation group. The gene designated as MSF1 has been subcloned on a 2.2-kilobase pair fragment and its nucleotide sequence determined. The predicted protein product of MSF1 has a molecular weight of 55,314 and has several domains of high primary sequence homology to the alpha subunit of the Escherichia coli phenylalanyl-tRNA synthetase. Based on the phenotype of G120 mutants and the homology to the bacterial protein, MSF1 is proposed to code for the alpha subunit of yeast mitochondrial phenylalanyl-tRNA synthetase. Disruption of the chromosomal copy of MSF1 in the respiratory-competent haploid strain W303 1B induces a phenotype similar to G120 mutants but does not affect cell viability, indicating that the cytoplasmic phenylalanyl-tRNA synthetase of yeast is encoded by a separate gene. Although the E. coli and yeast mitochondrial aminoacyl-tRNA synthetases are sufficiently similar in their primary sequences to suggest a common evolutionary origin, they have undergone significant changes as evidenced by the low homology in some regions of the polypeptide chains and the presence in the mitochondrial enzyme of two domains that are lacking in the bacterial phenylalanyl-tRNA synthetase. PMID- 3029121 TI - Studies on the synthesis of sterol carrier protein-2 in rat adrenocortical cells in monolayer culture. Regulation by ACTH and dibutyryl cyclic 3',5'-AMP. AB - The effects of ACTH or dibutyryl cyclic AMP (Bt2cAMP) on the synthesis of sterol carrier protein-2 (SCP2) have been studied in rat adrenocortical cells in monolayer culture. Radiolabeling of total cellular proteins with [35S]methionine and immunoprecipitation with antibodies directed against rat liver SCP2, followed by polyacrylamide gel electrophoresis and fluorography, showed a 3-4-fold increase in the rate of synthesis of SCP2 in cells treated for 48 h with ACTH (1 microM) or Bt2cAMP (0.1 mM). The induction of SCP2 synthesis depended upon the concentrations of ACTH or Bt2cAMP with an ED50 of 8 and 100 nM, respectively, and increased linearly with time between 12 and 48 h of treatment. Immunoprecipitation of SCP2 synthesized in a rabbit reticulocyte in vitro translation system programmed with RNA isolated from cells treated with ACTH or Bt2cAMP revealed increased synthesis of SCP2 compared to RNA from control cells. The immunoprecipitable rat adrenal SCP2, synthesized in a cell-free translation system, showed mobility corresponding to Mr of 14,400 upon sodium dodecyl sulfate polyacrylamide gel electrophoresis and was clearly larger than immunodetectable SCP2 synthesized in cultured adrenal cells (Mr = 11,300). The electrophoretic mobilities of rat liver SCP2 synthesized in cultured cells and in a cell-free translation system were the same as the respective forms from rat adrenal. It is concluded that the synthesis of SCP2 in rat adrenocortical cells is induced by ACTH and that the induction is mediated by cAMP and may involve increased levels of translatable mRNA encoding a higher molecular weight precursor form of SCP2, which presumably undergoes post-translational processing yielding the mature form. PMID- 3029122 TI - cDNA cloning and expression in Escherichia coli of a plasminogen activator inhibitor from human placenta. AB - Two nearly full-length cDNAs for placental plasminogen activator inhibitor (PAI) have been isolated from a human placenta lambda gt11 cDNA library. One positive (lambda PAI-75.1) expressed a protein that could adsorb and purify anti-PAI antibodies. The expressed protein inhibited the activity of human urokinase in a fibrin autography assay, and formed a 79-kDa (reduced) covalent complex with 125I urokinase that could be immunoprecipitated with anti-PAI. The cDNA insert of the longer isolate (lambda PAI-75.15) consisted of 1909 base pairs, including a 5' noncoding region of 55 base pairs, an open reading frame of 1245 base pairs, a stop codon, a 3'-noncoding region of 581 base pairs, and a poly(A) tail. The size of the mRNA was estimated to be 2.0 kilobases by Northern blot analysis. The translated amino acid sequence consisted of 415 amino acids, corresponding to a 46.6-kDa protein. The sequence was related to members of the serpin gene family, particularly ovalbumin and the chicken gene Y protein. Like these avian proteins, placental PAI appears to lack a cleavable NH2-terminal signal peptide. Residues 347-376 were identical to the partial sequence reported recently for a PAI isolated from the human monocytic U-937 cell line. Placental PAI mRNA was apparently expressed at low levels in human umbilical vein endothelial cells, but was not detectable in HepG2 hepatoma cells. It was present in U-937 cells and was inducible at least 10-fold by phorbol 12-myristate 13-acetate. Thus placental PAI is a unique member of the serpin gene family, distinct from endothelial-type PAI. It is probably identical to monocyte-macrophage PAI. PMID- 3029123 TI - The effect of supercoil and temperature on the recognition of palindromic and non palindromic regions in phi X174 replicative form DNA by S1 and Bal31. AB - The effect of supercoil and temperature on the topology of phi X174 replicative form (RF) DNA was studied using single-strand specific endonucleases S1 and Bal31 as probes for cruciform extrusion and other structural perturbations of the B helix. Both enzymes were found to recognize specifically and reproducibly over 30 sites, most of which were cleaved by both enzymes independent of the superhelicity of the genome. A negative superhelical density exceeding 0.06 stabilized a transition in the DNA conformation that generated several new cleavage sites for Bal31. The underlying structures appeared to be only transiently stable and were lost from in vitro supercoiled DNA during brief incubation at 65 degrees C. They were generally absent from in vivo supercoiled RF DNA of equal superhelicity as a consequence of the extraction and storage procedure. Mapping of the cleavage sites suggested that they were preferentially located near the beginnings and ends of genes and that the structural basis for at least some of them was the extrusion of relatively small palindromes into the cruciform state. Insertion of a short synthetic palindromic sequence into the phi X174 genome generated a supercoil-dependent, temperature-sensitive secondary structure that was cleaved in the Bal31 but not the S1 reaction, further supporting the hypothesis that even small cruciforms with stem size of 7 or less base pairs may be transiently stable. Subjecting supercoiled RF DNA to the typical S1 reaction conditions induced a topological shift that diminished all but one of the supercoil-induced Bal31 recognition sites and promoted the formation of one major new site. PMID- 3029124 TI - Promoter-glutathione S-transferase Ya cDNA hybrid genes. Expression and conferred resistance to an alkylating molecule in mammalian cells. AB - Promoter-glutathione S-transferase Ya cDNA hybrid genes were constructed and analyzed to determine the efficiency with which the Ya coding sequence was transcribed and also to determine the associated levels of Ya-specific enzyme activity in mammalian cells which had received the hybrid gene constructs via electroporation. Promoter-containing fragments from either the SV40 early region or the herpes simplex thymidine kinase gene were positioned 5' to the Ya cDNA present in the pGTB38 plasmid. Both promoters supported transcription in in vitro run-off incubations containing a rat cell extract. Efficient transcription was also observed in both monkey Cos cells and mouse C3H/10T1/2 cells. Constructs containing the SV40 promoter and a residual portion of the homopolymeric G tail used in the original Ya cDNA cloning consistently gave 4-50-fold higher levels of transcript than other promoter-cDNA configurations. Associated with transcription of the hybrid gene was the appearance of a glutathione S-transferase YaYa specific enzyme activity (delta 5-androstene-3,17-dione isomerization) in cytosols of cells electroporated with the hybrid genes. 50-260-fold increases in Ya-specific enzyme activity were found in Cos or C3H/10T1/2 cells containing multiple, episomal copies of the plasmid constructs; enzyme levels dropped in cells containing fewer, integrated plasmid copies. When a mixed population of Cos cells containing YaYa overexpressing cells was treated with benzo(a)pyrene (+/-) anti-diol epoxide, a cytotoxic alkylating molecule and known YaYa substrate, a 20 30-fold enrichment in clones of YaYa overexpressing cells was seen among those cells which survived the treatment. The results clearly indicate that glutathione S-transferase isozymes can be overexpressed in mammalian cells and that this is accompanied by significant biological resistance to a known alkylating molecule. PMID- 3029125 TI - Adult forms of the Ca2+ATPase of sarcoplasmic reticulum. Expression in developing skeletal muscle. AB - Two separate genes encode fast-twitch and slow-twitch/cardiac muscle forms of the Ca2+ ATPase of sarcoplasmic reticulum. Full length Ca2+ ATPase clones have been isolated from adult rabbit fast-twitch, slow-twitch, and cardiac muscles. Segments of these clones containing unique sequences have been used as probes to study developmental changes in Ca2+ ATPase transcripts. The fast-twitch Ca2+ ATPase transcript undergoes developmentally regulated alternative splicing in which a penultimate 42-base pair exon is retained in the adult transcript but is excised in the neonatal transcript. This additional exon shifts the exon encoding the neonatal carboxyl-terminal sequence, -Asp-Pro-Glu-Asp-Glu-Arg-Arg-Lys (Brandl, C. J., Green, N. M., Korczak, B., and MacLennan, D. H. (1986) Cell 44, 597-607) into a nontranslated region and results in the expression of an adult isoform with a carboxyl-terminal -Gly. The neonatal form of the fast-twitch Ca2+ ATPase represents 72% of the fast-twitch Ca2+ ATPase transcripts just prior to birth but only 17% by 14 days of age and 4% in adult fast-twitch muscle. Adult slow-twitch, adult cardiac, and neonatal skeletal muscles express an identical Ca2+ATPase mRNA transcript which is distinct from either of the fast-twitch forms. The slow-twitch/cardiac Ca2+ ATPase is the predominant form expressed in late fetal and early neonatal rabbit skeletal muscle, but this form is lost as the skeletal muscle differentiates into a fast-twitch state. Three or more alternative polyadenylation signals exist for this mRNA in all tissues with the most 3' signal predominating. PMID- 3029126 TI - Carcinogens with diverse mutagenic activities initiate neoplastic guinea pig cells that acquire the same N-ras point mutation. AB - N-ras has been identified by molecular cloning and DNA sequence analysis as the activated oncogene in carcinogen-induced guinea pig transformation. The deduced guinea pig amino acid sequence differs from that of human and mouse by 1 and 4 residues, respectively; the mismatches were in the C-terminal half of the fourth exon. The activated N-ras clone has an AT to TA transversion at the third position of codon 61 which results in the insertion of histidine instead of glutamine. The same activated N-ras gene with the identical mutation was found in all lines regardless of initiating carcinogen (aromatic aryl hydrocarbons or alkylating agents). These results suggest that the mutational event was independent of the mutagenic activity of the initiating carcinogen. PMID- 3029127 TI - Molecular cloning and characterization of cDNA for human myeloperoxidase. AB - Partial amino acid sequence of human myeloperoxidase was determined, and a 41 base oligonucleotide containing deoxyinosines at four positions was chemically synthesized. By using the oligonucleotide as a probe, cDNA clones for human myeloperoxidase were isolated from a cDNA library constructed with mRNA from human promyelocytic leukemia HL-60 cells. One of the clones containing a 2.6 kilobase insert was subjected to nucleotide sequence analysis. The sequence was found to contain an open reading frame, 2,235 nucleotides coding for a protein of 745 amino acids with a calculated Mr of 83,868. The heavy chain of myeloperoxidase, consisting of 467 amino acids, was located on the COOH terminus half of the protein. The RNA specified by the cDNA was prepared using SP6 RNA polymerase and translated in rabbit reticulocyte lysates, and the product was identified as human myeloperoxidase by immunoprecipitation with rabbit anti-human myeloperoxidase antibody. By Northern hybridization analysis of RNA from leukemic cells, it was shown that myeloperoxidase mRNA is abundantly expressed in human promyelocytic HL-60 and mouse myeloid leukemia NFS-60 cells. Furthermore, the results of Southern hybridization analysis of human genomic DNA suggest that there are one or two genes for myeloperoxidase in the human haploid genome. PMID- 3029128 TI - The structure of the rat glutathione S-transferase P gene and related pseudogenes. AB - We have isolated the rat placental-type glutathione S-transferase (GST-P) gene from a lambda phage library using GST-P cDNA clone, pGP5 (Sugioka, Y., Kano, T., Okuda, A., Sakai, M., Kitagawa, T., and Muramatsu, M. (1985) Nucleic Acids Res. 13, 6049-6057), as a probe. The rat GST-P gene is about 3 kilobase pairs long and contains 7 exons and 6 introns, encoding the same GST-P protein specified by pGP5. The cap site maps 70 nucleotides upstream from the translation initiation site. The canonical promoter "TATA" box was found 27 base pairs upstream from the putative cap site. Two hundred base pairs upstream from the cap site are rich in G + C residues (61%), and the hexanucleotide sequence 5'-GGGCGG-3' is found at position -47 to -42. We have also isolated several processed-type pseudogenes which were presumably originated by reverse transcription followed by insertion at target sites. PMID- 3029129 TI - Unique positioning of reconstituted nucleosomes occurs in one region of simian virus 40 DNA. AB - A direct end label method was used to study the positioning of nucleosome arrays on several long (greater than 2200 base pairs) SV40 DNA fragments reconstituted in vitro with core histones. Comparison of micrococcal nuclease cutting sites in reconstituted and naked DNA fragments revealed substantial differences in one DNA region. When sufficient core histones were annealed with the DNA to form closely spaced nucleosomes over most of the molecule, a uniquely positioned array of four nucleosomes could be assigned, by strict criteria, to a 610-base pair portion of the SV40 "late region," with a precision of about +/- 20 base pairs. In some other DNA regions, a number of alternative nucleosome positions were indicated. The uniquely positioned four-nucleosome array spanned the same 610 nucleotides on two different DNA fragments that possessed different ends. Removal of a DNA region that had contained a terminal nucleosome of the array, by truncation of the fragment before reconstitution, did not affect the positioning of the other three nucleosomes. As the core histone to DNA ratio was lowered, evidence for specific positioning of nucleosomes diminished, except within the region where the four uniquely positioned nucleosomes formed. This region, however, does not appear to have a higher affinity for core histones than other regions of the DNA. PMID- 3029130 TI - Mutants of Saccharomyces cerevisiae defective in sn-1,2-diacylglycerol cholinephosphotransferase. Isolation, characterization, and cloning of the CPT1 gene. AB - A colony autoradiographic assay for the sn-1,2-diacylglycerol cholinephosphotransferase activity in Saccharomyces cerevisiae was developed. Twenty-two mutants defective in cholinephosphotransferase activity were isolated. Genetic analysis revealed that all of these mutations were recessive, and three complementation groups were identified. The cholinephosphotransferase activities in membranes prepared from cpt1 mutants were reduced 2-10-fold compared to wild type activity. The cholinephosphotransferase activities of two cpt1 isolates differed from wild-type activity with respect to their apparent KM for CDP choline. The residual cholinephosphotransferase activities of cpt1 isolates were more sensitive to inhibition by CMP than the wild-type activity. The CPT1 gene was cloned by genetic complementation of cpt1 using a yeast genomic library. In strains transformed with the CPT1-bearing plasmid, a 5-fold overproduction of cholinephosphotransferase activity with wild-type kinetic properties was observed. The CPT1 gene was localized to a 1.2-2.4-kilobase region of DNA by transposon Tn5 mutagenesis and deletion mapping. An insertional mutant of the CPT1 gene was constructed and introduced into the chromosome by integrative transformation. The resulting cpt insertional mutant fell into the cpt1 complementation group. The cholinephosphotransferase activity in membranes prepared from the cpt1 insertional mutant was reduced 5-fold and exhibited CMP sensitivity. The sn-1,2-diacylglycerol ethanolaminephosphotransferase activities in membranes from all of the cpt1 isolates including the insertional mutant were normal. The data indicate that the cloned CPT1 gene represents the yeast cholinephosphotransferase structural gene, that the yeast choline- and ethanolaminephosphotransferase activities are encoded by different genes, and that the CPT1 gene is nonessential for growth. PMID- 3029131 TI - Chemoattractant-elicited increases in Dictyostelium myosin phosphorylation are due to changes in myosin localization and increases in kinase activity. AB - We previously reported (Berlot, C. H., Spudich, J. A., and Devreotes, P. N. (1985) Cell 43, 307-314) that cAMP stimulation of chemotactically competent Dictyostelium amoebae causes transient increases in phosphorylation of the myosin heavy chain and 18,000-dalton light chain in vivo and in vitro. In this report we investigate the mechanisms involved in these changes in phosphorylation. In the case of heavy chain phosphorylation, the amount of substrate available for phosphorylation appears to be the major factor regulating the in vitro phosphorylation rate. Almost all heavy chain kinase activity is insoluble in Triton X-100, and the increase in the heavy chain phosphorylation rate in vitro parallels an increase in Triton insolubility of myosin. Changes in heavy chain phosphatase activity are not involved in the changes in the in vitro phosphorylation rate. In the case of light chain phosphorylation, increases in the vitro phosphorylation rate occur under conditions where the amount of substrate available for phosphorylation is constant and phosphatase activity is undetectable, implicating light chain kinase activation as the means of regulation. The specificity of the myosin kinases operating in vivo and in vitro was explored using phosphoamino acid and chymotryptic phosphopeptide analysis. The light chain is phosphorylated on serine both in vivo and in vitro, and phosphopeptide maps of the light chain phosphorylated in vivo and in vitro are indistinguishable. In the case of the heavy chain, both serine and threonine are phosphorylated in vivo and in vitro, although the cAMP-stimulated increases in phosphorylation occur primarily on threonine. Phosphopeptide maps of the heavy chain show that the peptides phosphorylated in vitro represent a major subset of those phosphorylated in vivo. The kinetics of the transient increases in myosin phosphorylation rates observed in vitro can be predicted quantitatively from the in vivo myosin phosphorylation data assuming that there is a constant phosphatase activity. PMID- 3029132 TI - Autonomic interplay in VIP-induced tracheal smooth muscle relaxation. AB - VIP-induced inhibitory responses of the guinea-pig tracheal pouch, an in vivo preparation designed to demonstrate non-noradrenergic non-cholinergic innervation, where determined in chloraloseurethane anaesthetized animals under control conditions, or pretreated with atropine, propranolol, both atropine and propranolol or atropine-propranolol with indomethacin. Of the five groups, the control group showed a significantly greater cumulative dose-response relaxation to VIP than did the treatment groups. Among the treatment groups, the propranolol pretreated animals showed significantly greater relaxation than the remaining treatments. The group that received atropine alone and the combination of atropine and propranolol showed the least relaxation to VIP. The relaxations in these two groups did not differ significantly. The significantly greater relaxation of the pouch to VIP in the propranolol treated group suggested that the non-noradrenergic inhibitory mechanism for VIP is dependent upon the existing smooth muscle tone of the pouch. Pretreatment of the guinea-pigs with indomethacin did not cause a significant change in the relaxation of the pouch due to VIP suggesting that VIP caused airway smooth muscle relaxation by a mechanism other than that of the release of bronchodilator prostaglandins. PMID- 3029133 TI - Possible interaction of cholinergic nerves with two different (pre and post) sites of the neuromuscular junction in guinea-pig vas deferens. AB - Effects of various drugs on the mechanical responses of the longitudinal smooth muscle of guinea-pig vas deferens evoked by ACh and field nerve stimulation were examined. ACh (28-280 microM) produced a contraction consisting of two phases. The first phase of the contraction was suppressed by guanethidine and by nicotinic antagonists. The second phase was suppressed only by atropine. Both phases were unaffected by TTX or prazosin. Field stimulation (0.1 msec, 40 Hz) evoked contractions which also consisted of two (early and late) phases. Guanethidine (0.1-1 microM) suppressed both phases whilst prazosin (1 microM) suppressed only the late phase. Atropine (0.1 microM) suppressed both phases whilst physostigmine (5 microM) potentiated both phases of field stimulation evoked contractions. Pentolinium suppressed both phases of field stimulation evoked contractions at low concentrations (2-10 microM), but potentiated them at a higher concentration (100 microM). dTC at a low concentration (0.5 microM) suppressed the early phase, but slightly enhanced the late phase of field stimulation responses. At a higher concentration (20 microM), dTC potentiated both phases of the response. Pentolinium and dTC did not affect the contractions induced by 90 mM-K ions, ATP or NA. These results suggest that cholinergic nerves possess an excitatory action not only directly at the smooth muscle but also at the noradrenergic nerve terminals. The role of each receptor is discussed further. PMID- 3029134 TI - Pharmacological studies on the role of cholinergic nerves in the neuromuscular transmission in the circular smooth muscle of guinea-pig vas deferens. AB - Effects of various drugs were examined on the mechanical responses of the circular smooth muscle of guinea-pig vas deferens evoked by field stimulation and ACh. During field stimulation (0.1 msec, 3-80 Hz) a contraction consisting of two (initial and second) phases occurred. In addition, another contraction (after response) occurred after the cessation of the stimulation particularly at higher frequencies (40-80 Hz). Guanethidine (2-20 microM) suppressed both the initial and second phases of the field stimulation-evoked responses. Prazosin (0.1-1 microM) suppressed the second phase but minimally affected the initial phase. Both phases were suppressed by atropine (0.1-1 microM) and potentiated by physostigmine (5 microM). dTC (0.5-20 microM) did not suppress either phase but potentiated the second phase. ACh (0.28-280 microM) produced a contraction, which was reduced by atropine but was little affected by either dTC or TTX. These results suggest that at the postsynaptic site of the neuromuscular junction in this tissue the sympathetic process activates the smooth muscle through two different types of receptor; one is non-noradrenergic and the other is an alpha adrenoreceptor. In addition, a cholinergic process also activates the smooth muscle through muscarinic receptors. PMID- 3029135 TI - Treatment of winged scapula by pectoralis major transfer. AB - We report the transfer of the sternal part of the pectoralis major to the lower pole of the scapula in 15 patients with winged scapula. At follow-up after 1 to 16 years nine had a satisfactory result and were gainfully employed, though in four of these re-operation had been necessary. Two patients had fair results; the transplant functioned, but they had limited voluntary control. Four were failures: two had had paralysis of other shoulder girdle muscles in addition to the serratus anterior. The indications for the operation are discussed. PMID- 3029136 TI - Levels of active metabolites of vitamin D3 in the callus of fracture repair in chicks. AB - The levels of the active metabolites of vitamin D were measured in the callus and in the epiphyseal growth plate of chicks given radioactive cholecalciferol during fracture healing. Those levels were correlated with the histological findings. Three groups of chicks were studied: a control group with no fracture, chicks with fractures fixed by Kirschner wire, and chicks with unfixed fractures. A significant increase in the levels of the active metabolites was found in the callus during the first few days after fracture. The levels of 25 hydroxycholecalciferol [25(OH)D3] and of 24,25-dihydroxycholecalciferol [24,25(OH)2D3] were higher when there was no fixation, while those of 1,25 dihydroxycholecalciferol [1,25(OH)2D3] were higher after fixation. The concentrations of these metabolites in the proximal epiphysis of the tibia were similar to those found in the callus. Based on these findings it is suggested that the active metabolites of vitamin D are directly involved in the process of fracture repair. PMID- 3029137 TI - Interaction of anthracycline antibiotics with human neutrophils: superoxide production, free radical formation and intracellular penetration. AB - Human neutrophils exposed to 10(-4) M doxorubicin and the derivatives epirubicin and thepirubicin revealed a different intracellular penetration and distribution pattern as demonstrated by fluorescence microscopy and fluorimetric determination of drug intracellular concentration. While doxorubicin was found to be a potent inducer of superoxide generation from resting cells, epirubicin exhibited less superoxide-inducing power. Thepirubicin on the contrary did not show any superoxide-inducing effect. Moreover the anthracyclines tested all inhibited the phorbol ester-stimulated chemiluminescent response to the same extent, which suggested a common target for the drug action. Anthracycline-stimulated superoxide production seems to correlate with the cardiotoxic effects. The most cardiotoxic drug, doxorubicin, is the most potent inducer of superoxide generation, while epirubicin, which is less cardiotoxic, has a relatively limited effect on superoxide production. Thepirubicin which has been shown not to induce delayed cardiomyopathy has no effect on superoxide release from the cells. PMID- 3029138 TI - Establishment, growth properties, and morphological characteristics of permanent human small cell lung cancer cell lines. AB - Cell lines from SCLC were established with a success rate of 43% from different metastatic sites of treated and untreated patients. All 6 SCLC cell lines grew as floating cell aggregates without substrate adherence. The degree of aggregation ranged from very tight spheroids to very loose sheets and chains. This gross morphological property showed a striking correlation to the PDT, with short PDTs in loose growing cell lines and long PDTs in tight growing cell lines. Cell size and nuclear features, i.e., chromatin pattern and nucleolar prominence, also seemed to correlate with the PDT and gross morphology. All SCLC cell lines had dense core granules by electron microscopical examination. Several different serum-free and serum-supplemented growth media were tested for their feasibility in establishing and permanently growing SCLC. Serum-free SIT medium and SIT2.5 medium provided the best results in liquid culture. For semisolid SCLC cultivation, R 10 medium was superior to all other media tested. These cell lines are currently under intensive biochemical, molecular biological, and cytogenetical investigation in different laboratories and thus provide a tool for studying the biology of lung cancer. PMID- 3029139 TI - Nucleotide sequence comparison of the predicted first exonic region of goldfish ras gene between normal and neoplastic tissues. AB - The first exonic and its flanking regions of a ras-related gene in cultured erythrophoroma cells and normal liver of goldfish (Carassius auratus) were sequenced and their sequences compared. The two samples demonstrated identical sequences for the Alu I fragment of 245 nucleotides, of which 70 nucleotides might correspond to the 5'-upstream segment and 64 to the first intervening sequence, indicating that the predicted exonic region encodes from the 1st to the 37th amino acids from the N-terminal of mammalian ras protein. The length of the predicted first exon of goldfish was the same as that of mammals. There was a 5 nucleotide difference in this exonic region (length 111 nucleotides) from another previously determined ras-related sequence of normal goldfish. This difference was much smaller than that between any two of the three mammalian ras genes. Enhanced expression of ras gene was observed in erythrophoroma cells, although it was uncertain which ras gene was responsible for generation of the observed RNA. PMID- 3029140 TI - Microinjection of gelsolin into living cells. AB - Gelsolins are actin-binding proteins that cap, nucleate, and sever actin filaments. Microinjection of cytoplasmic or plasma gelsolin into living fibroblasts and macrophages did not affect the shape, actin distribution, deformability, or ruffling activity of the cells. Gelsolin requires calcium for activity, but the NH2-terminal half is active without calcium. Microinjection of this proteolytic fragment had marked effects: the cells rounded up, stopped ruffling, became soft, and stress fibers disappeared. These changes are similar to those seen with cytochalasin, which also caps barbed ends of actin filaments. Attempts to raise the cytoplasmic calcium concentration and thereby activate the injected gelsolin were unsuccessful, but the increases in calcium concentration were minimal or transient and may not have been sufficient. Our interpretation of these results is that at the low calcium concentrations normally found in cells, gelsolin does not express the activities observed in vitro at higher calcium concentrations. We presume that gelsolin may be active at certain times or places if the calcium concentration is elevated to a sufficient level, but we cannot exclude the existence of another molecule that inhibits gelsolin. Microinjection of a 1:1 gelsolin/actin complex had no effect on the cells. This complex is stable in the absence of calcium and has capping activity but no severing and less nucleation activity as compared with either gelsolin in calcium or the NH2 terminal fragment. The NH2-terminal fragment-actin complex also has capping and nucleating activity but no severing activity. On microinjection it had the same effects as the fragment alone. The basis for the difference between the two complexes is unknown. The native molecular weight of rabbit plasma gelsolin is 82,500, and the extinction coefficient at 280 nm is 1.68 cm2/mg. A new simple procedure for purification of plasma gelsolin is described. PMID- 3029141 TI - Microinjection of ubiquitin: intracellular distribution and metabolism in HeLa cells maintained under normal physiological conditions. AB - Radioiodinated ubiquitin was introduced into HeLa cells by erythrocyte-mediated microinjection. Subsequent electrophoretic analyses revealed that the injected ubiquitin molecules were rapidly conjugated to HeLa proteins. At equilibrium, 10% of the injected ubiquitin was conjugated to histones and 40% was distributed among conjugates of higher molecular weight. Although the remaining ubiquitin molecules appeared to be unconjugated, the free pool of ubiquitin decreased by one-third and additional conjugates were present when electrophoresis was performed at low temperature under nonreducing conditions. Molecular weights of these labile conjugates suggest that they are ubiquitin adducts in thiolester linkage to activating enzymes. Despite the fairly rapid degradation of injected ubiquitin (t1/2 approximately 10-20 h), the size distribution of ubiquitin conjugates within interphase HeLa cells remained constant for at least 24 h after injection. The intracellular locations of ubiquitin and ubiquitin conjugates were determined by autoradiography, by differential sedimentation of subcellular fractions in sucrose, and by extraction of injected cells with buffer containing Triton X-100. Free ubiquitin was found mostly in the cytosolic or Triton X-100 soluble fractions. As expected, histone conjugates were located predominately in the nuclear fraction and exclusively in the Triton X-100-insoluble fraction. Although high molecular weight conjugates were enriched in the Triton X-100 insoluble fraction, their size distribution was similar to that of soluble conjugates. When injected HeLa cells were exposed to cycloheximide to inhibit protein synthesis, the size distribution of ubiquitin conjugates was similar to that found in untreated cells. Moreover, high molecular weight conjugates decreased less than 20% after inhibition of protein synthesis. These results indicate that most ubiquitin conjugates are not newly synthesized proteins which have been marked for destruction. PMID- 3029142 TI - Microinjection of ubiquitin: changes in protein degradation in HeLa cells subjected to heat-shock. AB - Ubiquitin was radiolabeled by reaction with 125I-Bolton-Hunter reagent and introduced into HeLa cells using erythrocyte-mediated microinjection. The injected cells were then incubated at 45 degrees C for 5 min (reversible heat shock) or for 30 min (lethal heat-shock). After either treatment, there were dramatic changes in the levels of ubiquitin conjugates. Under normal culture conditions, approximately 10% of the injected ubiquitin is linked to histones, 40% is found in conjugates with molecular weights greater than 25,000, and the rest is unconjugated. After heat-shock, the free ubiquitin pool and the level of histone-ubiquitin conjugates decreased rapidly, and high molecular weight conjugates predominated. Formation of large conjugates did not require protein synthesis; when analyzed by two-dimensional electrophoresis, the major conjugates did not co-migrate with heat-shock proteins before or after thermal stress. Concomitant with the loss of free ubiquitin, the degradation of endogenous proteins, injected hemoglobin, BSA, and ubiquitin was reduced in heat-shocked HeLa cells. After reversible heat-shock, the decrease in proteolysis was small, and both the rate of proteolysis and the size of the free ubiquitin pool returned to control levels upon incubation at 37 degrees C. In contrast, neither proteolysis nor free ubiquitin pools returned to control levels after lethal heat shock. However, lethally heat-shocked cells degraded denatured hemoglobin more rapidly than native hemoglobin and ubiquitin-globin conjugates formed within them. Therefore, stabilization of proteins after heat-shock cannot be due to the loss of ubiquitin conjugation or inability to degrade proteins that form conjugates with ubiquitin. PMID- 3029143 TI - Analyses of the interactions between retinoid-binding proteins and embryonal carcinoma cells. AB - [3H]Retinoic acid (RA) and [3H]retinol bind in an unsaturable manner to isolated nuclei from Nulli-SCC1 and PCC4.aza1R embryonal carcinoma (EC) cells. When nuclei are challenged with the same labeled retinoids on their respective binding proteins (CRABP and CRBP), much less binding is observed and the binding is saturable. RA-CRABP does not compete with [3H]retinol-CRBP for binding to specific Nulli-SCC1 nuclear sites, whereas retinol-CRBP (but not apo-CRBP) actually potentiates the binding of [3H]RA-CRABP to these nuclei. The binding of [3H]RA-CRABP and [3H]retinol-CRBP is not dramatically affected by prior removal of the outer nuclear membrane with Triton X-100. However, treatment with the detergent after the binding reaction is complete removes about half of the bound [3H]RA-CRABP and almost all of the bound [3H]retinol-CRBP. We measured specific retinoid-binding activities in nucleoplasmic extracts of Nulli-SCC1 and PCC4.aza1R cells. The only readily detectable specific binding activity in nucleoplasmic extracts from untreated cells was for [3H]retinol in PCC4.aza1R preparations. Nucleoplasmic extracts from Nulli-SCC1 and PCC4.aza1R cells pretreated with RA had considerable levels of specific [3H]RA-binding activity with little or no increase in [3H]retinol binding. By contrast, similar extracts from Nulli-SCC1 cells treated with retinol bound large amounts of both [3H]retinol and [3H]RA. Under the same conditions, PCC4.aza1R extracts also contained [3H]RA-binding activity with no increase in [3H]retinol binding above the high endogenous levels. Although these results might reflect translocation of binding proteins from cytoplasm to nucleus, other interpretations must be considered since we often observed an increase, rather than the expected reduction, in cytoplasmic retinoid-binding protein levels. PMID- 3029145 TI - Role of Na-K ATPase in regulation of resting membrane potential of cultured rat skeletal myotubes. AB - The role of Na-K ATPase in the determination of resting membrane potential (Em) as a function of extracellular K ion concentration was investigated in cultured rat myotubes. The Em of control myotubes at 37 degrees C varied as a function of (K+)0 with a slope of about 58-60 mV per ten-fold change in (K+)0. Inhibition of the Na-K pump with ouabain or by reduced temperature revealed that this relation consists of two components. One, between (K+)0 of 10 and 100 mM, remains unchanged by alterations in enzyme activity; The second, between (K+)0 of 1 and 10 mM, is related to the amount of Na-K pump activity, the slope decreasing as pump activity decreases. Indeed, with complete inhibition of the Na-K pump, Em does not change over the range of (K+)0 1 to 10 mM. Measurements of 86Rb efflux and input resistance of individual myotubes showed that membrane permeability does not change as (K+)0 increases from 1 to 10 mM but increases as (K+)0 increases further. Monensin, which increases Na ion permeability, increases Em at values of external K+ below 10 mM, and is without effect at higher values of K+ concentration. The effect of monensin is blocked by ouabain. Tetrodotoxin, which blocks voltage-dependent Na+ channels, decreases Em at low (2-10 mM) K+. We conclude that changes in Em as a function of extracellular K+ concentration in the physiological range are not adequately explained by the diffusion potential hypothesis of Em, and that other theories (electrogenic pump, surface-absorption) must be considered. PMID- 3029146 TI - Graded amplification of the Na,K-ATPase across a subclonal series: effects on membrane physiology. AB - We have generated a series of clonally related cell lines which differ in the level of amplified expression of the Na,K-ATPase. These lines, originally derived from the ouabain resistant HeLa variant C+, expressed different numbers of binding sites for the Na,K-ATPase inhibitor ouabain, ranging from 2.9 X 10(6)/cell to 11.8 X 10(6)/cell. Amplification of the genes for both subunits of the enzyme was also seen but was not strictly correlated with level of expression. The influxes of histidine and tetraphenylphosphonium were measured across a series, including HeLa S3 and revertants, expressing from 0.74 X 10(6) to 10.5 X 10(6) ouabain-binding sites per cell. Tetraphenylphosphonium influx rate, presumed to be a function of membrane potential, varied linearly with ouabain binding site number, while histidine influx varied with the log of ouabain binding site number. Our results suggest that membrane potential increases in a simple fashion across our series of amplified lines. However, histidine influx was unaffected by treatments which cause membrane depolarization and a decrease in tetraphenylphosphonium influx rate. We propose that increasing histidine influx rates across our amplified series reflects exchange acceleration of L system transport due to increased intracellular pools of L system reactive amino acids. The Na,K-ATPase is ultimately responsible for most active transport across the plasma membrane. The consistent, graded physiological alterations seen across this series of closely related lines, chosen for graded enzyme expression, demonstrate the value of this novel genetic approach to the study of the energization of membrane transport. PMID- 3029144 TI - Reconstitution of transport of vesicular stomatitis virus G protein from the endoplasmic reticulum to the Golgi complex using a cell-free system. AB - Transport of the vesicular stomatitis virus-encoded glycoprotein (G protein) between the endoplasmic reticulum (ER) and the cis Golgi compartment has been reconstituted in a cell-free system. Transfer is measured by the processing of the high mannose (man GlcNAc2) ER form of G protein to the man5GlcNAc5 form by the cis Golgi enzyme alpha-mannosidase I. G protein is rapidly and efficiently transported to the Golgi complex by a process resembling that observed in vivo. G protein is trimmed from the high mannose form to the man5GlcNAc2 form without the appearance of the intermediate man GlcNAc2 oligosaccharide species, as is observed in vivo. G protein is found in a sealed membrane-bound compartment before and after incubation. Processing in vitro is sensitive to detergent, and the Golgi alpha-mannosidase I inhibitor 1-deoxymannorjirimycin. Transport between the ER and Golgi complex in vitro requires the addition of a high speed supernatant (cytosol) of cell homogenates, and requires energy in the form of ATP. Efficient reconstitution of export of protein from the ER requires the preparation of homogenates from mitotic cell populations in which the nuclear envelope, ER, and Golgi compartments have been physiologically disassembled before cell homogenization. These results suggest that the high efficiency of transport observed here may require reassembly of functional organelles in vitro. PMID- 3029147 TI - Phosphorylation of regulatory subunit of type I cyclic AMP-dependent protein kinase: biphasic effects of cyclic AMP in intact S49 mouse lymphoma cells. AB - Intact S49 mouse lymphoma cells were used as a model system to study the effects of cyclic AMP (cAMP) and its analogs on the phosphorylation of regulatory (R) subunit of type I cAMP-dependent protein kinase. Phosphorylation of R subunit was negligible in mutants deficient in adenylate cyclase; low levels of cAMP analogs, however, stimulated R subunit phosphorylation in these cells to rates comparable to those in wild-type cells. In both wild-type and adenylate cyclase-deficient cells, R subunit phosphorylation was inhibited by a variety of N6-substituted derivatives of cAMP; C-8-substituted derivatives were generally poor inhibitors. Two derivatives that were inactive as kinase activators (N6-carbamoylmethyl-5' AMP and 2'-deoxy-N6-monobutyryl-cAMP) were also ineffective as inhibitors of R subunit phosphorylation. Preferential inhibition by N6-modified cAMP analogs could not be ascribed simply to selectivity for the more aminoterminal (site I) of the two cAMP-binding sites in R subunit: Analog concentrations required for inhibition of R subunit phosphorylation were always higher than those required for activation of endogenous kinase; 8-piperidino-cAMP, a C-8-substituted derivative that is selective for cAMP-binding site I, was relatively ineffective as in inhibitor; and, although thresholds for activation of endogenous kinase by site I-selective analogs could be reduced markedly by coincubation with low levels of site II-selective analogs, no such synergism was observed for the inhibitory effect. The uncoupling of cyclic nucleotide effects on R subunit phosphorylation from activation of endogenous protein kinase suggests that, in intact cells, activation of cAMP-dependent protein kinase requires more than one and fewer than four molecules of cyclic nucleotide. PMID- 3029148 TI - Deoxyribonucleotide metabolism and cyclic AMP resistance in hydroxyurea-resistant S49 T-lymphoma cells. AB - We investigated the cell cycle regulation of deoxyribonucleoside triphosphate (dNTP) metabolism in hydroxyurea-resistant (HYUR) murine S49 T-lymphoma cell lines. Cell lines 10- to 40-fold more hydroxyurea-resistant were selected in a stepwise manner. These HYUR cells exhibited increased CDP reductase activity (5- to 8-fold) and increased dNTP pools (up to 5-fold) that appeared to result from increased activity of the M2 subunit (binding site of hydroxyurea) of ribonucleotide reductase. These characteristics remained stable when the cells were grown in the absence of hydroxyurea for up to 2 years. In both wild type and hydroxyurea-resistant cell populations synchronized by elutriation, dCTP and dTTP pools increased in S phase, whereas dATP and dGTP pools generally remained the same or decreased, suggesting that allosteric effector mechanisms were operating to regulate pool sizes. Additionally, CDP reductase activity measured in permeabilized cells increased in S phase in both wild type and hydroxyurea resistant cells, suggesting a nonallosteric mechanism of increased ribonucleotide reductase activity during periods of active DNA synthesis. While wild type S49 cells could be arrested in the G1 phase of the cell cycle by dibutyryl cyclic AMP, hydroxyurea-resistant cell lines could not be arrested in the G1 phase by exogenous cyclic AMP or agents that elevate the concentration of endogenous cyclic AMP. These data suggest that cyclic AMP-generated G1 arrest in S49 cells might be mediated by the M2 subunit of ribonucleotide reductase. PMID- 3029149 TI - Transforming growth factor-alpha attenuates the acquisition of aromatase activity by cultured rat granulosa cells. AB - The effect of transforming growth factor-alpha (TGF alpha) on granulosa cell differentiation, as assessed by the acquisition of aromatase activity, was evaluated in vitro by using a primary culture of rat granulosa cells. Harvested from immature, diethylstilbestrol-treated rats, granulosa cells were cultured under serum-free conditions for 72 hr in the presence of saturating concentrations (10(-7)M) of aromatase substrate androstenedione with or without the specific experimental agents. Basal aromatase activity, as assessed by the generation of radioimmunoassayable estrogen was negligible, remaining unaffected by treatment with TGF alpha (10 ng/ml) by itself. Whereas treatment with follicle stimulating hormone (FSH) resulted in a substantial increase in the extent of aromatization, concurrent treatment with TGF alpha (10 ng/ml) resulted in significant (P less than 0.05), yet reversible inhibition (78 +/- 5.6%) of FSH action. Significantly, this effect of TGF alpha could not be accounted for by a decrease in cellular viability or plating efficiency nor by a decrease in the number of cells or their DNA content. Although independent of the FSH dose employed, the TGF alpha effect proved dose- and time-dependent, with an apparent median inhibitory dose (EC50) of 0.33 +/- 0.04 ng/ml, and a minimal time requirement of 48 hr. Capable of substantial inhibition of the forskolin stimulated accumulation of extracellular adenosine 3', 5' cyclic monophosphate (cAMP) and estrogen, TGF alpha had a measurable albeit limited effect on N6, 2-'O Dibutyryladenosine 3':5'-cyclic monophosphate-supported estrogen production. Relative potency comparison revealed epidermal growth factor (EGF; EC50 = 0.24 +/ 0.03 ng/ml) and TGF alpha to be virtually equipotent as regards the attenuation of FSH-stimulated estrogen biosynthesis. Taken together, our findings indicate that TGF alpha, like EGF, acting at subnanomolar concentrations, is capable of attenuating the FSH-stimulated (but not basal) accumulation of estrogen. This effect of TGF alpha proved time- and dose-dependent, involving virtually complete neutralization of FSH action at site(s) both proximal and distal to cAMP generation. As such, these findings provide yet another example of the remarkable qualitative and quantitative similarities between EGF and TGF alpha, thereby reaffirming the prospect that ligands of the EGF/TGF alpha receptor may play a modulatory role in the course of granulosa cell ontogeny. PMID- 3029150 TI - Site-specific mutagenesis of cDNA clones expressing a poliovirus proteinase. AB - The cleavage of poliovirus precursor polypeptides occurs at specific amino acid pairs that are recognized by viral proteinases. Most of the polio-specific cleavages occur at glutamine-glycine (Q-G) pairs that are recognized by the viral encoded proteinase 3C (formerly called P3-7c). In order to carry out a defined molecular genetic study of the enzymatic activity of protein 3C, we have made cDNA clones of the poliovirus genome. The cDNA region corresponding to protein 3C was inserted into an inducible bacterial expression vector. This recombinant plasmid (called pIN-III-C3-7c) utilizes the bacterial lipoprotein promoter to direct the synthesis of a precursor polypeptide that contains the amino acid sequence of protein 3C as well as the amino- and carboxy-terminal Q-G cleavage signals. These signals have been previously shown to allow autocatalytic production of protein 3C in bacteria transformed with plasmid pIN-III-C3-7c. We have taken advantage of the autocatalytic cleavage of 3C in a bacterial expression system to study the effects of site-specific mutagenesis on its proteolytic activity. One mutation that we have introduced into the cDNA region encoding 3C is a single amino acid insertion near the carboxy-terminal Q-G cleavage site. The mutant recombinant plasmid (designated pIN-III-C3-mu 10) directs the synthesis of a bacterial-polio precursor polypeptide that is like the wild-type construct (pIN-III-C3-7c). However, unlike the wild-type precursor, the mutant precursor cannot undergo autocatalytic cleavage to generate the mature proteinase 3C. Rather, the precursor is able to carry out cleavage at the amino terminal Q-G site but not at the carboxy-terminal site. Thus, we have generated an altered poliovirus proteinase that is still able to carry out at least part of its cleavage activities but is unable to be a suitable substrate for self cleavage at its carboxy-terminal Q-G pair. PMID- 3029152 TI - Gas chromatographic method for the determination of fluconazole, a novel antifungal agent, in human plasma and urine. PMID- 3029151 TI - Direct analytical and preparative resolution of enantiomers using albumin adsorbed to silica as a stationary phase. AB - A rapid and simple method for the preparation of high-performance liquid chromatography columns for chiral separations is described. The stationary phase is prepared by adsorbing bovine serum albumin on silica. Both analytical and preparative applications are described. A polarimeter was used as a detector to determine the enantiomer elution orders. PMID- 3029153 TI - Determination of sulbactam in biological fluids by high-performance liquid chromatography. PMID- 3029154 TI - Gonadotropin-releasing hormone (GnRH)-binding sites in human breast cancer cell lines and inhibitory effects of GnRH antagonists. AB - GnRH-binding sites have previously been described in human breast tumors, and a GnRH agonist has been shown to inhibit growth of the MCF-7 human breast cancer cell line. We have investigated the presence of GnRH-binding sites in ZR-75-1, MDA-MB-231, Sk Br 3, MDA-MB-157, and MCF-7 human breast cancer cell lines and the effect of GnRH analogs on the incorporation of [3H]thymidine and 14C-labeled amino acids into DNA and protein. Specific GnRH-binding sites were present in membrane preparations of all five human breast carcinoma cell lines. Studies in three cell lines indicated low affinity (Kd, 1.6-3.0 X 10(-6) M) GnRH binding similar to that reported in human placenta and corpus luteum. In contrast, human pituitary GnRH receptors were of high affinity (Kd, 4.8 X 10(-9) M). Breast carcinoma cell GnRH-binding sites also differed from the pituitary receptor in their inability to discriminate between GnRH and superactive analogs. Binding of a [125I]GnRH analog to ZR-75-1 breast cancer cells and pituitary membranes was affected similarly by various cations. GnRH antagonists rapidly inhibited [3H]thymidine incorporation into DNA (within 3 hr), and this effect was reversible. GnRH antagonists also inhibited cell growth, but only after 6 days. GnRH agonists did not alter either thymidine incorporation or growth. The present observations of low affinity GnRH-binding sites in breast cancer cell lines and inhibitory effects of GnRH antagonists point to the possibility of an autocrine regulatory role of GnRH-like peptides in mammary cells. PMID- 3029155 TI - The effect of mevinolin on steroidogenesis in patients with defects in the low density lipoprotein receptor pathway. AB - Adrenal steroidogenesis is dependent upon cholesterol derived from both de novo biosynthesis and uptake of plasma lipoproteins through the low density lipoprotein (LDL) receptor pathway. Recent studies have demonstrated that patients homozygous for familial hypercholesterolemia have a mild impairment in cortisol secretion during maximal ACTH stimulation. Mevinolin, a competitive inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase, has been used clinically to inhibit de novo cholesterol synthesis in patients with familial hypercholesterolemia. In this study we examined hypothalamic-pituitary-adrenal function in seven patients with defects in the LDL receptor pathway, both before and during treatment with oral mevinolin (20 mg, twice a day), to assess whether inhibition of cholesterol synthesis influences steroidogenesis under basal conditions and in response to ovine CRH and exogenous ACTH. Two months after initiation of therapy, high density lipoprotein cholesterol levels were significantly elevated, and LDL cholesterol levels were reduced, although not normalized. Basal and ovine CRH-stimulated adrenocortical function were normal in all patients both before and during therapy. Plateau plasma cortisol concentrations achieved during maximal ACTH stimulation were lower than those in control subjects in all patients both before and during therapy. All patients, however, had an approximately 3-fold increase over basal values. These results suggest that treatment of patients with defects in the LDL receptor pathway with mevinolin improves the plasma lipid profile and does not result in adrenal dysfunction or further exacerbation of the mild impairment of adrenal function during maximal ACTH stimulation. PMID- 3029157 TI - The dissociation of renin and aldosterone during critical illness. AB - A syndrome of elevated PRA accompanied by inappropriately low plasma aldosterone (ALDO) levels has been identified in some critically ill patients. To determine whether this phenomenon is due to a disturbance in factors that stimulate ALDO, we measured PRA, angiotensin II (AII), potassium (K+), and ACTH levels in 83 patients admitted to an intensive care unit. In 59 patients, PRA was greater than 2.0 ng/ml X h. Of these, 24 had an ALDO to PRA ratio (ALDO/PRA) below 2 (group I), and 35 had an ALDO/PRA ratio of 2 or more (group II). An ALDO/PRA ratio below 2 was deemed inappropriately low. Despite markedly elevated PRA [34 +/- 12 (+/- SE) ng/ml X h], the group I patients had inappropriately low ALDO levels (19 +/- 5 ng/dL). Patients in group II had significantly higher ALDO levels (48 +/- 6 ng/dL) despite lower PRA (9 +/- 1 ng/ml X h). AII levels were appropriately elevated in group I (39 +/- 26 pg/mL) and significantly greater (P less than 0.5) than those in group II. PRA correlated well with AII in both groups. There were no differences in plasma ACTH or K+ in these 2 groups, and plasma cortisol levels were similarly elevated in both groups of patients. Of 66 consecutively studied patients, 14 (21%) had inappropriate ALDO (group I). Mortality was significantly greater in group I (75%) than in group II (46%; P less than 0.001). In summary, a significant subset (21%) of seriously ill patients have inappropriately low ALDO levels despite elevated PRA. This dissociation is not due to an impairment of AII production or changes in plasma ACTH or K+. This phenomenon is associated with a higher mortality during critical illness. In light of evidence of decreased adrenal androgen secretion during severe illness, this dissociation of renin and aldosterone may represent an additional adrenal adaptation designed to promote cortisol production in critically ill patients. PMID- 3029156 TI - Defects in insulin binding and autophosphorylation of erythrocyte insulin receptors in patients with syndromes of severe insulin resistance and their parents. AB - Genetic forms of severe insulin resistance are often characterized by alterations in binding and/or kinase properties of the insulin receptor. To evaluate whether alterations in insulin receptor kinase of erythrocytes can be used as genetic markers, we studied patients with two apparently inherited conditions of severe insulin resistance (leprechaunism and the type A syndrome of insulin resistance) and their families. In the two propositi, [125I]insulin binding to intact erythrocytes was decreased by 64% and 45%, respectively. This was primarily due to a decrease in receptor number and was found in intact cells and solubilization of the receptors. Similar insulin binding defects were found on monocytes. Insulin-stimulated tyrosine kinase activity of the solubilized receptor from erythrocytes was also decreased and to a similar extent as binding. Parents of neither patient had clinical manifestations of leprechaunism or the type A syndrome. Furthermore, no alterations in insulin receptor binding or kinase activity were found in erythrocytes from the mothers. Insulin binding in the father of the type A patient was also normal, whereas the father of the leprechaun had decreased receptor affinity. Receptors extracted from the both fathers' cells had a 40-60% decrease in maximal insulin-stimulated phosphorylation and significant rightward shifts of the insulin dose-response curves (ED50, 141 and 42 ng/mL, respectively; control ED50, 16 ng/mL). The finding of biochemical defects in insulin receptor kinase activity in clinically unaffected parents of patients suggests that these alterations may be useful genetic markers and more sensitive than insulin binding studies for studying pattern of inheritance of these diseases. PMID- 3029158 TI - Genetic defects of steroidogenesis in premature pubarche. AB - Twenty three patients (19 girls, 4 boys) presented with typical features of premature pubarche between the ages of 2-7 yr. The patients were studied for the presence of an adrenal steroidogenic defect by ACTH stimulation testing (Cortrosyn, 0.25 mg iv bolus dose). Based on published nomogram standards for serum 17-hydroxyprogesterone (17-OHP), seven patients (30%) were diagnosed as having the nonclassical symptomatic form of 21-hydroxylase deficiency [mean post ACTH 4244 +/- 1113 (SD) ng/dl]. Three patients (13%) were diagnosed to have a mild form of 3 beta-hydroxysteroid dehydrogenase deficiency based upon the response of serum delta 5-17-hydroxypregnenolone (delta 5-17P) and dehydroepiandrosterone, and the ratio of delta 5-17P/17-OHP to ACTH stimulation (delta 5-17P: 1543 +/- 272 ng/dl vs. Tanner stage I control subjects, 350 +/- 197 ng/dl; dehydroepiandrosterone: 675 +/- 190 ng/dl vs. Tanner stage I control subjects, 82 +/- 79 ng/dl; delta 5-17P/17-OHP: 8.1 +/- 2.6 vs. Tanner stage I control subjects, 1.4 +/- 0.6). No enzyme defect could be identified in the remaining 13 patients (57%). Eleven patients with premature pubarche, with and without an adrenal enzymatic defect, underwent dexamethasone suppression. In all patients the measured steroid levels were suppressed. Thus, the dexamethasone suppression test alone did not distinguish the pathogenesis of premature pubarche. In conclusion, premature pubarche is more commonly due to a partial enzyme defect in adrenal steroidogenesis than has been previously recognized. PMID- 3029159 TI - Sex hormone-binding globulin secretion by human hepatocarcinoma cells is increased by both estrogens and androgens. AB - The secretion of sex hormone-binding globulin (SHBG) by the human hepatocarcinoma cell line Hep G2 was increased significantly not only by estradiol (E2) but also by testosterone (T), dihydrotestosterone, and the synthetic androgen danazol in the presence, but not the absence, of fetal calf serum. The secretion of SHBG also was stimulated by the antiestrogen tamoxifen, although this required the use of longer incubation periods and higher concentrations than were required for the steroids. The antiandrogen cyproterone acetate and cortisol (5 microM) decreased SHBG secretion. Pregnanediol (5 microM) had no discernible effect. These changes were considered specific as none of the compounds altered either the secretion of total protein by the cells or their total protein content. Cells incubated with a mixture of E2 (0.5 microM) and T (0.5 microM) secreted significantly more SHBG than did cells incubated with E2 (1.0 microM), indicating these steroids exert their effects through different mechanisms. The increases with E2 and T were reduced significantly by tamoxifen and cyproterone acetate, respectively, suggesting receptor mediation of the steroid effects. The E2-related increase was abolished by cycloheximide, indicating that the changes were due to alterations in the synthesis of SHBG rather than to alterations in the release of previously synthesized protein. These findings suggest the T-related decrease in plasma SHBG levels may be due to causes other than a decrease in the hepatic synthesis of the protein. Additionally, they indicate that Hep G2 cells may prove suitable for examining the regulation of the SHBG gene by a variety of compounds. PMID- 3029160 TI - Trophoblastic disease: a retrospective view. PMID- 3029162 TI - Analysis by RNA-RNA hybridization assay of intertypic rotaviruses suggests that gene reassortment occurs in vivo. AB - Antigenic characterization of human and animal rotaviruses by the plaque reduction neutralization assay has shown the existence of naturally occurring intertypes. Antiserum to M37, a rotavirus strain isolated from an asymptomatic neonate, neutralizes both Wa and ST3 strains, which are classified as serotype 1 and serotype 4 human rotaviruses, respectively. Likewise, antiserum to SB-1A, a porcine rotavirus, neutralizes rotavirus strains belonging to serotype 4 or 5. Plaque reduction neutralization assay of reassortant rotaviruses produced in vitro from these intertypes indicates that these viruses share one antigenically related outer capsid protein, VP3, with one serotype and another antigenically related outer capsid protein, VP7, with the other serotype. Thus, M37 is related to ST3 on the basis of its fourth-gene product, VP3, and to Wa on the basis of its ninth-gene product, VP7, whereas SB-1A is related to Gottfried (serotype 4 porcine rotavirus) via VP7 and to OSU (serotype 5 porcine rotavirus) via VP3. RNA RNA hybridization studies revealed a high degree of homology between the VP3 or VP7 gene segments responsible for shared serotype specificity. Thus, the fourth gene segments of M37 and ST3 were highly homologous, while M37 and Wa had homology between their ninth gene segments. SB-1A and Gottfried were homologous not only with respect to the ninth gene but had complete homology in all other genes except the fourth gene. The fourth gene of SB-1A was highly homologous with the fourth gene of OSU. These observations suggested that SB-1A was a naturally occurring reassortant between Gottfried-like and OSU-like porcine rotavirus strains. Our observations also suggested that intertypes may result from genetic reassortment in nature. PMID- 3029163 TI - Genomic variation of adenovirus type 5 isolates recovered from bone marrow transplant recipients. AB - We characterized the genomic variation of adenovirus type 5 isolates recovered from bone marrow transplant recipients in Seattle between 1976 and 1982. By restriction endonuclease analysis, we identified three new adenovirus genomic variants, each associated with a single invasive adenovirus infection. In addition, we were able to obtain suggestive evidence for a nosocomial spread of a particular group of isolates within this population. This study demonstrates that the technique of restriction endonuclease analysis is an important epidemiological tool for investigating viral infections. PMID- 3029161 TI - Physicochemical characterization of porcine pararotavirus and detection of virus and viral antibodies using cell culture immunofluorescence. AB - A cell culture immunofluorescence (CCIF) assay was optimized for detection of porcine pararotavirus (group C rotavirus) in intestinal contents. The greatest viral infectivity was observed when MA104 cells (5 days after subculturing) were rinsed and refed in serum-free medium before inoculation, pancreatin was added to the inocula, and the inocula were centrifuged onto the cells. Gentamicin treatment of pararotavirus samples to reduce bacterial contamination also reduced the viral infectivity of these samples for MA104 cells. An indirect CCIF assay was used to determine the prevalence of pararotavirus and rotavirus antibodies in pig sera. In pigs from four herds, pararotavirus antibodies were detected in 100% (68 of 68) of adults and 59% (24 of 41) of weanling pigs, while 86% (24 of 28) of nursing pigs from 12 herds had pararotavirus antibodies. The physicochemical properties of pararotavirus were examined and compared with those of group A rotaviruses by using the CCIF assay to quantitate in vitro changes in viral infectivity. Pararotavirus was inactivated (greater than or equal to 99% reduction in titer) by heating to 56 degrees C for 30 min, was slightly labile at pH 3 (16 to 34% reduction in titer), and was stable at pH 5 (0 to 17% reduction in titer) and in either (3 to 19% reduction in titer). One group A rotavirus (Gottfried strain) was stable at 56 degrees C (0% reduction in titer), whereas the OSU strain of group A rotavirus was inactivated at this temperature (99% reduction in titer). PMID- 3029164 TI - Prevalence of antibody to group B (atypical) rotavirus in humans and animals. AB - Enzyme-linked immunosorbent assays were developed for the detection of group B rotavirus antigen and antibody. The specificities of both assays were evaluated for antigens and serum specific for rotavirus groups A to D. Serum collected in the United Kingdom from different animal species exhibited the following high prevalence of group B rotavirus-specific antibody: pigs, 97%; cattle, 71%; sheep, 91%; and goats, 91%. In human serum, a lower prevalence of group B-specific antibody was detected; serum from blood donors showed 10% prevalence, and serum from veterinarians showed 4% prevalence. PMID- 3029165 TI - Capillary enzyme immunoassay for rapid detection of herpes simplex virus in clinical specimens. AB - Capillary enzyme immunosorbent assay (CapELISA) is a modification of the standard enzyme immunosorbent assay which permits rapid detection of viral antigens in clinical specimens. The capillary tube format provides a very large reactive surface relative to the sample size. The close proximity of antigen to antibody in the tube optimizes the reaction, resulting in increased sensitivity and shorter incubation requirements. Sensitivity is further enhanced by use of a biotin-avidin enzyme detector system and a fluorogenic rather than a colorigenic substrate. The assay is performed at ambient temperature and requires less than 2 h. It is read with an inexpensive hand-held black light. Data for the use of CapELISA for the detection of herpes simplex virus in clinical specimens are presented. The results show greater than or equal to 85% sensitivity and 100% specificity when compared with tissue culture tests on the same samples. This new system should be advantageous for the diagnosis, treatment, and management of patients with herpesvirus infections and for the prevention of neonatal herpes acquired by passage of the fetus through an infected birth canal. PMID- 3029166 TI - Diagnosis of herpes simplex virus by direct immunofluorescence and viral isolation from samples of external genital lesions in a high-prevalence population. AB - One hundred thirty specimens from 108 consecutive patients with a history of recurrent genital ulcerations were used in a study comparing herpes simplex virus (HSV) isolation with a direct fluorescent-antibody (DFA) technique using mouse monoclonal antibodies. HSV was isolated from 70% of vesicular lesions, 67% of pustular lesions, 32% of ulcerative lesions, and 17% of crusted lesions, whereas the DFA technique detected HSV antigen in 87, 67, 30, and 10% of lesions in similar stages, respectively. When both methods were used, HSV was identified in 97, 79, 45, and 17% of vesicles, pustules, ulcers, and crusted lesions, respectively. The overall sensitivity and specificity of the DFA technique in comparison with virus isolation (VI) were 74 and 85%, respectively. Of the 17 patients from whom DFA-positive, VI-negative samples were obtained, HSV was subsequently isolated from a genital lesion in 14, suggesting that they were not DFA false-positives. Similarly, of the 46 patients whose initial lesion samples were DFA and VI negative, 37 (80%) had HSV identified from subsequent genital lesions during follow-up. Thus, a single sample for VI or DFA testing from a recurrent genital lesion had a sensitivity of only 53 and 51%, respectively. Combining the DFA technique and VI increased the sensitivity of laboratory diagnosis of a single recurrent episode of genital HSV; however, repetitive laboratory testing was often required to confirm the diagnosis of recurrent genital HSV infection. PMID- 3029167 TI - Age-related infections with rotavirus, rotaviruslike virus, and atypical rotavirus in turkey flocks. AB - The genome electropherotyping technique was used in longitudinal surveys to detect group A rotavirus, rotaviruslike virus (RVLV), atypical rotavirus (ATR), and reovirus in intestinal contents or fecal specimens collected from turkeys in 10 commercial and 2 research station flocks. These viruses were detected in turkeys from 8 to 10 commercial flocks surveyed. Of 278 specimens collected from turkeys less then 29 days old in commercial flocks, 79 (28.4%) contained one or more viruses, whereas only 1 of 120 specimens collected from turkeys older than 28 days had virus. Viruses were detected in commercial turkeys between 3 and 35 days old, and over a third of the specimens collected from birds during their first week of life were positive for group A rotavirus. Between 8 and 28 days of age, commercial turkeys were infected with group A rotavirus, RVLV, ATR, and reovirus. ATR was the only virus detected in birds older than 28 days. Overall, group A rotavirus and RVLV were each detected in 39 specimens, and ATR was detected in 7 specimens; reovirus was detected in 2 specimens. Eight of the positive specimens contained two viruses. All 102 specimens collected from turkeys 1 to 56 days old in the two research station flocks were negative for virus. PMID- 3029168 TI - Efficiency of immunofluorescence for rapid detection of common respiratory viruses. AB - Rapid immunofluorescence (FA) methods for the detection of common respiratory viruses were compared with culture results over a 3-year period to assess the relative efficiency of FA in a clinical laboratory setting. For respiratory syncytial virus, efficiencies were high (sensitivity, 90 to 95%; specificity, 92 to 95%). The sensitivity of FA for detection of parainfluenza virus type 1, parainfluenza virus type 3, influenza A virus, and adenoviruses ranged from 28 to 63%, but specificities for these viruses were uniformly 98 to 100%. The observations form a basis for consideration of selective reduction of routine culture procedures for specimens with initial positive rapid FA results; however, the possibility of dual viral infection in some situations must also be considered. PMID- 3029169 TI - Immunoglobulin level in donor blood reactive for antibodies to human immunodeficiency virus. AB - Blood samples from 98 asymptomatic volunteer blood donors, including 55 that were reactive for antibodies to human immunodeficiency virus (HIV) in Western blot (WB) assay, were tested for levels of immunoglobulin G (IgG), IgM, and titer of antibodies to HIV, cytomegalovirus, and herpes simplex virus. Levels of IgG were significantly elevated (P less than or equal to 0.001) in donors with specific anti-HIV reactivity. A total of 69% of donors with anti-HIV had IgG levels of greater than or equal to 12 mg/ml, and 44% had IgG levels of greater than or equal to 14.5 mg/ml. Levels of IgM were not significantly different among WB reactive and nonreactive donors. The titer of anti-HIV was significantly (P less than 0.02) correlated with IgG levels among donors reactive in the WB assay. Elevation of IgG, however, was not significantly associated with the presence of anticytomegalovirus or anti-herpes simplex virus antibodies. The data show that elevation of IgG may represent an early manifestation of HIV infection before the development of clinical symptoms of acquired immunodeficiency syndrome. PMID- 3029170 TI - Detection of St. Louis encephalitis virus antigen in mosquitoes by capture enzyme immunoassay. AB - Surveillance methods that measure St. Louis encephalitis (SLE) virus activity in nature may provide forewarning of its epidemic occurrence in humans. An antigen capture enzyme immunoassay was developed to detect SLE virus in infected mosquitoes. The assay detected purified SLE viral antigen at a concentration of 62 pg/0.1 ml when antigen was incubated overnight; 250 pg/0.1 ml was detected in a single-day assay (antigen incubated for 3 h). The assay detected 67.9 and 70.8% of laboratory-prepared pools of infected mosquitoes after 3 h and overnight incubation, respectively. The sensitivity of the procedure was 90.5% in identifying pools with infectious titers greater than dex 3.0. The specificity of the assay was controlled by retesting positive pools preincubated with SLE virus and normal antibodies, which led to a diminution of signal in the pools containing viral antigen. The procedure was suitably specific in discriminating between SLE and related flaviviruses, detecting only high infectious doses of heterologous antigens. PMID- 3029171 TI - Membrane filtration enzyme immunoassay, a novel, rapid method for measurement of virus-specific immunoglobulins G and M and detection of viral antigens. AB - A novel membrane filtration enzyme immunoassay (MF EIA) is described in which virus-antibody complexes formed are trapped onto the surface of membranes with low protein-binding affinity by vacuum filtration. Class-specific immunoglobulin G (IgG) or IgM antibody was measured by adding enzyme-conjugated antiimmunoglobulin and incubating prior to the final wash and addition of enzyme substrate. Influenza A virus-specific IgG antibodies measured by MF EIA showed similar sensitivity for detecting seroconversion in volunteers administered influenza virus subunit vaccines and subtype specificity comparable to that observed by the hemagglutination inhibition technique. Cytomegalovirus-specific IgG antibodies measured by MF EIA with commercially available complement fixation antigens gave results similar to those of conventional enzyme-linked immunosorbent assay and complement fixation tests. The MF EIA method is also suitable for detection of rotavirus antigen in feces. PMID- 3029172 TI - Evaluation of three immunofluorescence assays for culture confirmation and typing of herpes simplex virus. AB - Three pairs of monoclonal antibodies, supplied in kits by Electro-Nucleonics, Inc. (ENI), The Syva Co., and Kallestad Laboratories, Inc. (KL), were evaluated for the laboratory confirmation and typing of herpes simplex virus (HSV). Of 108 coded HSV slide preparation, run in parallel with each monoclonal-antibody set, 103 were equivalent by the immunofluorescence assays. Among the five discordant isolates, three (2.8%) did not type with the KL monoclonal antibodies and two (1.9%) false-positive results occurred with the Syva typing system. All of the HSV clinical isolates tested were correctly typed with the ENI indirect immunofluorescence antigen detection system. Typing confirmation of the five discordant HSV isolates was performed by differential sensitivity to 5-bromo-2' deoxyuridine and endonuclease cleavage analysis of the viral DNA. Use of the Syva and KL direct immunofluorescence antigen detection systems for the identification of HSV isolates is less time-consuming than use of the ENI indirect antigen detection system; however, sensitivity and specificity may be lost. PMID- 3029173 TI - Early testing of cell cultures for detection of hemadsorbing viruses. AB - Hemadsorption of primary monkey kidney cell cultures was commenced at 1 day after inoculation to evaluate how rapidly influenza A virus, influenza B virus, and parainfluenza virus types 1, 2, and 3 could be detected by this method from respiratory specimens. Overall, 38% of all isolates could be detected within 24 h of inoculation, and 69% could be detected within 48 h. All influenza A viruses and all but one influenza B virus were detected by day 3. PMID- 3029174 TI - Centrifugation-shell vial technique for rapid detection of herpes simplex virus cytopathic effect in Vero cells. AB - A centrifugation-shell vial technique for herpes simplex virus detected the virus in 50 (24.6%) of 203 specimens by cytopathic effect at 48 h, compared with detection of the virus in 47 (23.2%) specimens by immunofluorescence at 24 h. The rapid detection of cytopathic effect may be useful to laboratories that are not interested in typing all herpes simplex virus specimens but that wish to reduce the cost of herpesvirus cultures. PMID- 3029175 TI - Phosphorylation protects neurofilaments against proteolysis. AB - During incubation with phosphatase, the 200 kDa neurofilament protein in cytoskeletal preparations is degraded extensively. Degradation, which is divalent cation-independent, does not occur when inhibitors of phosphatase are added. The 160 kDa chymotryptic fragment of neurofilaments or affinity-purified 200 kDa protein are not degraded by phosphatase. The results suggest that phosphorylated neurofilaments are protected against proteolysis, and dephosphorylated neurofilaments are degraded by a calcium-independent, endogenous proteinase which is associated with assembled neurofilaments or with other cytoskeletal components, and not with the phosphatase used. PMID- 3029176 TI - Viral specificity of multiple sclerosis tear immunoglobulins. AB - The mucosal immune system plays an important role in determining how an individual handles infection by most of the common viruses. Tears form part of this system and participate in mucosal immunity. We examined tears from multiple sclerosis (MS) and control subjects for antibodies to seven common neurotropic viruses. Antibodies to multiple viruses were detected in MS tears, especially when tears were obtained from chronic progressive active patients. In particular, MS patients showed a significantly greater prevalence of tear antibodies to measles, herpes simplex type I, and rubella virus. The tear anti-viral response included IgA as well as IgG antibody and appeared to be independent of serum. Mucosal fluid in MS patients, in addition to cerebrospinal fluid, also shows heightened humoral immune responses to multiple neurotropic viruses. PMID- 3029177 TI - Elevated adenosine deaminase activity and hereditary hemolytic anemia. Evidence for abnormal translational control of protein synthesis. AB - We have investigated the molecular basis of the marked elevation in erythrocyte adenosine deaminase (ADA) activity in a kindred with hereditary hemolytic anemia. Red cell ADA-specific activity was verified to be 70- to 100-fold normal levels. Western blots demonstrated a corresponding increase in erythrocyte ADA-specific immunoreactive protein. Analysis of genomic DNA revealed no evidence for amplification or major structural changes in the ADA gene. ADA-specific messenger RNA (mRNA) from proband reticulocytes was comparable in size and amount to mRNA from control reticulocytes. Translation of proband poly A+ reticulocyte mRNA in a rabbit reticulocyte lysate system and immunoprecipitation of 35S-labeled protein products with anti-ADA antibody yielded a band of approximately 42,000 apparent mol wt that was absent in translation products from control reticulocyte mRNAs. These data suggest that the increased ADA activity in red cells in this disorder results from the increased translation of an aberrant ADA mRNA. PMID- 3029178 TI - Heterogeneity of the factor IX locus in nine hemophilia B inhibitor patients. AB - DNA from nine hemophilia B patients who produce anti-factor IX inhibitors (antibodies), including two brothers, was analyzed by the Southern blotting method and hybridization with factor IX cDNA, intragenomic, and 3'-flanking probes. Two inhibitor patients were shown to have total deletions of the factor IX gene. Two other inhibitor patients, the brothers, were shown to have a presumably identical complex rearrangement of the factor IX gene involving two separate deletions. The first deletion is of approximately 5.0 kb and removes exon e. The second deletion is between 9 and 29 kb and removes exons g and h but leaves exon f intact. An abnormal Taq I fragment at one end of the deletion junctions acted as a marker for the inheritance of hemophilia B in the patients' family. Five other inhibitor patients have a structurally intact factor IX gene as detected by this method. Our studies indicate that whereas large structural factor IX gene defects predispose hemophilia B patients to developing an anti factor IX inhibitor, the development of an inhibitor can be associated with other defects of the factor IX gene. PMID- 3029181 TI - The therapy of genital warts. PMID- 3029179 TI - Mechanism for enhanced glucose transport response to insulin in adipose cells from chronically hyperinsulinemic rats. Increased translocation of glucose transporters from an enlarged intracellular pool. AB - The mechanism for the increased glucose transport response to insulin in adipose cells from chronically hyperinsulinemic rats was examined. Rats were infused with insulin s.c. for 2 wk. Isolated adipose cells were incubated with and without insulin, 3-O-methylglucose transport was measured, and glucose transporters in subcellular membrane fractions were assessed by cytochalasin B binding. Adipose cells from insulin-treated rats showed no change in basal but a 55% increase in insulin-stimulated glucose transport activity compared with those from control rats (7.1 +/- 0.8 vs. 4.6 +/- 0.5 fmol/cell per min, mean +/- SEM) and a corresponding increase in the concentration of glucose transporters in the plasma membranes (44 +/- 5 vs. 32 +/- 6 pmol/mg of membrane protein). In the low-density microsomes, glucose transporter concentrations in both basal and insulin stimulated states were the same, but the total numbers were greater in cells from the insulin-treated rats because of a 39% increase in low-density microsomal protein. Therefore, chronic experimental hyperinsulinemia in the rat enhances the stimulatory action of insulin on glucose transport in the adipose cell by increasing the concentration of glucose transporters in the plasma membranes. This results from an enlarged intracellular pool due to increased intracellular protein and enhanced glucose transporter translocation in response to insulin. PMID- 3029180 TI - Epidermal growth factor acts as a corticotropin-releasing factor in chronically catheterized fetal lambs. AB - Epidermal growth factor (EGF) has been reported to stimulate adrenocorticotropin hormone (ACTH), growth hormone and prolactin secretion from pituitary tissue in vitro, and in large doses evokes ACTH secretion in adult sheep in vivo. In order to assess a possible role for EGF in the pituitary hyperfunction characteristic of the in utero fetus, we measured changes in plasma immunoreactive ACTH concentrations after acute administration of saline, purified mouse EGF or synthetic ovine corticotropin releasing factor (CRF) to chronically catheterized fetal sheep. Both CRF and EGF were associated with increases in plasma immunoreactive ACTH concentrations. Peak values 60 min after 10-micrograms injections of either EGF or CRF increased from baseline ACTH values of 61 +/- 11 pg/ml to 191 +/- 37 and 178 +/- 25 pg/ml, respectively. Dose-response studies indicate that at low doses (less than 20 micrograms) EGF is as potent a stimulus for ACTH release as CRF. EGF infusion was not associated with detectable changes in circulating CRF, catecholamines, arginine vasopressin levels, or plasma growth hormone concentrations. We speculate that EGF may be important in the regulation of pituitary function in the developing mammalian fetus. PMID- 3029182 TI - Biopsy pathology of acquired immune deficiency syndrome (AIDS). AB - Between January 1982 and May 1986 279 biopsy specimens from 82 patients with acquired immune deficiency syndrome (AIDS) were examined. A wide variety of infectious conditions were diagnosed, the commonest being Pneumocystis pneumonia (n = 36), cytomegalovirus (n = 21), a variety of fungi (n = 8), mycobacteria (n = 7). Kaposi's sarcoma was the commonest tumour (n = 40), and there were two cases of extranodal lymphoma. Striking features were the unusual sites of disease and the occasional paucity of organisms. PMID- 3029183 TI - Modified latex agglutination test for anticytomegalovirus, suitable for pretransfusion screening. PMID- 3029184 TI - Sensitive in situ hybridisation technique using biotin-streptavidin-polyalkaline phosphatase complex. AB - A sensitive in situ hybridisation technique, using a biotin-streptavidin polyalkaline phosphatase complex detection system, was successfully applied to smears of fresh cultured cells, frozen sections, and formalin fixed paraffin processed tissue: the procedure was successful for DNA-DNA hybridizations using a variety of DNA probes. The detection method is rapid, reliable, and economical producing a purplish-blue precipitate at the site of hybridisation and clearly visible by low power light microscopy. PMID- 3029186 TI - Tests of the regenerative capacity of tectal efferent axons in the frog, Rana pipiens. AB - Experiments were designed to determine if neurons of the ranid optic tectum, a major target of the optic nerve, possess the same regenerative potential as optic axons. Normal tectal efferent (TE) projections were reexamined by using the anterograde transport of 3H-proline and autoradiography (n = 18), bulk-filling damaged TE axons with horseradish peroxidase (HRP; n = 18) and anterogradely transporting wheat germ agglutinin-HRP (n = 8) to label TE axons. Results were similar to reports that used degeneration methods (Rubinson: Brain Behav. Evol. 1:529-561, '68; Lazar: Acta. Biol. Hung. 20:171-183, '69). Following a brainstem hemisection just caudal to the nucleus isthmi (1-20 weeks), the ipsilateral descending TE pathway was autoradiographically examined (n = 20). While all other TE projections appeared normal, there was no detectable ipsilateral descending projection beyond the lesion site. Ascending TE axons were cut at the anterior tectal border by hemisecting the left diencephalon (LDH)--a lesion that also cuts optic axons projecting to the left tectum. There was no indication of TE axonal regeneration with the aid of autoradiography or HRP histochemistry 1-30 weeks postlesion (n = 48) even when the medial diencephalon was intentionally left intact (n = 4). However, in all four cases examined, optic axons regenerated following the same LDH where TE axonal regeneration failed (also see Stelzner, Lyon, and Strauss: Anat. Rec. 205:191A-192A, '83). Local effects of LDH should be similar for both the cut optic and cut TE axons. Other factors were tested that may contribute to the lack of TE axonal regeneration. Our results indicate that optic regeneration itself (n = 8), postaxotomy retrograde cell death of TE neurons (n = 6), deafferentation of the tectum of optic axons, and potential sprouting within tectal targets by intact contralateral TE axons (n = 10) are not critical factors aborting TE axonal regeneration. TE axons filled with HRP at chronic periods after LDH (n = 4) terminate anomalously near the LDH border. Many of these endings are similar to reactive endings or terminal clubs seen after axonal injury in the mammalian CNS. Our results suggest that this disparity in regenerative ability of optic and TE axons may be related to a difference in the responsive ability of these cell types to initiate or maintain axonal elongation after axotomy within the amphibian CNS environment. PMID- 3029185 TI - Cytochrome oxidase activity in the rat caudate nucleus: light and electron microscopic observations. AB - Cytochrome oxidase (CO) activity was examined in the neostriatum of normal adult rats at the light and electron microscopic level. At the light microscopic level a heterogeneous distribution of CO activity was observed and was characterized by patches of high activity ranging in size from 200 to 800 microns surrounded or adjacent to regions of lower activity. The most dorsomedial and ventromedial regions of the caudate nucleus appeared to be consistently high in activity in all animals. At the ultrastructural level CO reaction product was localized to the membranes and intracristal spaces of mitochondria. The most reactive mitochondria (those containing the denest precipitates of reaction product) were found within the dendrites of spiny neurons in all caudate regions. In areas of high CO activity the mitochondria within bundles of myelinated fibers and in many axon terminals were also highly reactive whereas those in neuronal somata, primary dendrites, and glial cells and processes exhibited relatively little activity. Quantitative study showed that mitochondria within dendrites accounted for most of the CO activity in caudate neuropil. The mitochondria within dendrites and axon terminals were more reactive in regions of high CO activity than in regions of low CO activity. No differences in the density of synapses or in the proportions of axospinous and axodendritic synapses were observed between CO-rich and CO-poor areas. Heterogeneity in the distribution of CO activity in the caudate nucleus may be related to the "patchy" pattern of localization previously observed for some neostriatal afferents, enzymes, transmitters, peptides, and receptor ligands.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3029187 TI - Development of synaptic connections between muscle sensory and motor neurons: anatomical evidence that postsynaptic dendrites grow into a preformed sensory neuropil. AB - The anatomical development of muscle sensory arbors and dendrites of brachial motoneurons in the spinal cord of the bullfrog was studied by labeling both types of cells with horseradish peroxidase. Sensory and motoneurons were labeled in tadpoles (stages XV-XVIII) by backfilling the triceps nerve in vivo with HRP throughout the stages in development when functional monosynaptic connections between these cells are first being formed. Individual triceps motoneurons were injected with HRP in other tadpoles at the same developmental stages. By stage XV, triceps sensory afferents already projected to and arborized in the ventral sensory neuropil region of the spinal cord where sensory-motor connections are made. In contrast, the dendrites of triceps motoneurons rarely were present in this region until stage XVI. By stage XVII, triceps dendrites in this region were common and they intermingled with the collaterals of muscle sensory axons. Thus, sensory axons supplying limb muscles grow into the future neuropil region well in advance of the arrival of motoneuronal dendrites. Electrophysiological studies have shown that the connections between triceps sensory and motor neurons are already specific at stage XVII, as soon as monosynaptic potentials between these cells can be detected (Frank and Westerfield: J. Physiol. (Lond.) 343:593-610, '83). The present anatomical results demonstrate that the processes of sensory and motor cells are not in close anatomical proximity before this time; thus the selection of appropriate synaptic partners must occur from the outset. PMID- 3029188 TI - The ventral striatopallidothalamic projection: I. The striatopallidal link originating in the striatal parts of the olfactory tubercle. AB - The projections from the striatal part of the olfactory tubercle were examined in rats, both with the aid of experimental silver impregnation methods following superficial laminar heat lesions of the tubercle and by the use of anterograde transport of Phaseolus vulgaris-leucoagglutinin (PHA-L) following injections of the lectin in the dense cell layer of the tubercle. Retrograde transport of fluorescent substances following injections of the tracer in the multiform layer of the tubercle were used to corroborate the results obtained by the anterograde transport and degeneration methods. The main and apparently only significant termination from the striatal cells in the olfactory tubercle is located immediately deep to the dense cell layer in areas that could be identified as part of the ventral pallidum on the basis of either the Nissl method or glutamate decarboxylase immunocytochemistry. Whereas a mediolateral topography is generally maintained by the ventral striatopallidal pathway originating in the dense cell layer, there is a considerable spread of the projection in the rostrocaudal direction. The dense projection field of the olfactory tubercle component of the ventral striatopallidal pathway permeates the ventrolateral part of the ventral pallidum, thereby complementing the termination of the accumbens projection to the more mediodorsal parts of the ventral pallidum. PMID- 3029190 TI - Coexistent pemphigus vulgaris and Paget's disease of the nipple. An immunohistochemical study. AB - We report the coexistence of pemphigus vulgaris and Paget's disease of the nipple in a patient with a documented history of pemphigus and a 2-year complaint of an eczematous breast lesion. A biopsy taken from the lesion showed the characteristic histologic features of both pemphigus and Paget's disease of the nipple. Immunohistochemical technics revealed intercellular IgG deposits characteristic of pemphigus and intracellular staining with antibody to epithelial membrane antigen in the cytoplasm of Paget's cells. This report is the first describing the coexistence of these two distinct disease entities in one tissue sample. This case underscores the necessity of early biopsy of recalcitrant nipple lesions even in the presence of another, well-documented skin disorder. PMID- 3029189 TI - The ventral striatopallidothalamic projection: II. The ventral pallidothalamic link. AB - The projection of ventral pallidal neurons to the mediodorsal nucleus of the thalamus (MD) was examined in rats by combined retrograde transport of horseradish peroxidase (HRP) after injections in the MD and glutamate decarboxylase (GAD) immunocytochemistry at light and electron microscopic levels, with and without prior exposure of the brains to colchicine. HRP was transported to the soma of medium-sized and large ventral pallidum neurons, which along with their long, large dendrites were contacted by many glutamate decarboxylase immunoreactive synaptic boutons. The retrograde tracer positive neurons bore a remarkable resemblance to the projecting cells of the globus pallidus and entopeduncular nucleus. When colchine exposure was included in the tissue preparation, some but not all tracer positive cells also exhibited cytoplasmic GAD immunoreactivity. PMID- 3029191 TI - Kaposi's sarcoma and acquired immunodeficiency syndrome. Postmortem findings in twenty-four cases. AB - Autopsy results on twenty-four patients with acquired immunodeficiency syndrome (AIDS) and Kaposi's sarcoma were reviewed. At postmortem, 29% of patients had evidence of visceral Kaposi's sarcoma without skin lesions. The most common sites for visceral involvement with Kaposi's sarcoma were as follows: lung (37%), gastrointestinal tract (50%), and lymph nodes (50%). At the time of death, only 25% of patients had evidence of cutaneous disease alone. The patients survived up to 36 months after the diagnosis of AIDS was made according to the specific diagnostic criteria established by the Centers for Disease Control. Many of the patients had severe, disseminated infections during the course of their AIDS illness. The most common infections diagnosed during the patients' clinical courses and/or at autopsy were cytomegalovirus (75%), candidiasis (50%), Mycobacterium avium intracellulare (50%), Pneumocystis carinii pneumonia (50%), bacterial pneumonia (33%), and herpes simplex virus (29%). Nearly 80% of the deaths were attributed to infection. In only one case was overwhelming Kaposi's sarcoma determined to be the cause of death. PMID- 3029194 TI - Eccrine syringofibroadenoma versus acrosyringeal nevus. PMID- 3029192 TI - Condylomata acuminata (genital warts). An epidemiologic view. AB - The understanding of condylomata acuminata (genital warts) has been enhanced by the recent development of diagnostic methods. Forty-two types of human papillomavirus were identified up to 1985, and at least sixteen types were involved in genital warts. The incidence of genital warts is around 0.1% in the general population and more than 0.5% in young persons. The incidence is known to be increasing rapidly and exceeding the incidence of genital herpes. Females are more prone to be affected. The epidemiologic evidence supporting the relationship between genital warts and genital cancer is overwhelming. The evidence also speaks for a strong correlation between genital warts and verrucous carcinoma of the genitalia, bowenoid papulosis, and laryngeal papilloma. The person having genital warts may also have other cutaneous warts. This observation is compatible with the finding that types of human papillomavirus involved in other cutaneous warts were found in genital warts. In view of the never and easier ways of diagnosing nonconspicuous condyloma acuminatum and the potential for malignant transformation of condyloma acuminatum, it is strongly recommended that patients should be followed up periodically for early detection of neoplasia. PMID- 3029193 TI - Successful treatment of glucagonoma-related necrolytic migratory erythema with dacarbazine. AB - A 63-year-old man with a glucagonoma syndrome is described. The diagnosis was confirmed by necrolytic migratory erythema, which is the most distinctive feature of the clinical syndrome. There was no chance of operative resection of the tumor because of liver metastases at the time of diagnosis. The patient was treated with dacarbazine. During this treatment the skin lesions disappeared completely. PMID- 3029195 TI - Bowenoid papulosis of the male and female genital tracts: risk of cervical neoplasia. PMID- 3029196 TI - CT and MR imaging of fatty tumors of the liver. AB - The presence of fat in hepatic masses narrows the range of differential diagnoses down to hepatic angiomyolipoma, lipoma, adenoma, hepatoma, metastatic fatty tumors of the liver, focal fatty infiltration of the liver, and extrahepatic fatty masses such as intraperitoneal implants from malignant teratomas, and packed omentum. We report six hepatic tumors containing fat (lipoma, hepatocellular carcinoma, and calcified mass with fat-fluid level) with CT and magnetic resonance (MR) imaging. The distribution of fat was diffuse in the lipomas and some hepatocellular carcinomas and localized in other hepatocellular carcinomas and fat-fluid masses. The density ranged from - 100 to 0 HU. High intensity areas on both T1- and T2-weighted MR images corresponded to the hypodense areas on CT. PMID- 3029198 TI - CT myelography of cauda equina actinomycosis. PMID- 3029197 TI - High resolution MR imaging of glomus tumor. AB - High resolution magnetic resonance images of a surgically confirmed glomus tumor of the finger obtained with a surface coil are shown. Magnetic resonance has the potential to preoperatively evaluate soft tissue tumors of these regions which are poorly imaged by other modalities. PMID- 3029200 TI - [Hydroxylapatite augmentation in maxillo-facial surgery]. PMID- 3029199 TI - Palisading cutaneous fibrous histiocytoma. AB - We report 6 cases of an unusual variant of cutaneous fibrous histiocytoma in which nuclear palisading is a prominent feature. The lesions were equally distributed between males and females with a widely variable age range. Half of the cases occurred on the digits. Histopathologically, the lesions were characterized by areas of nuclear palisading with formation of Verocay-like bodies in addition to the more typical features of the "fibrous" variant of cutaneous fibrous histiocytoma. The differential diagnostic features between these lesions and those of other tumors in which nuclear palisading is seen are discussed. PMID- 3029201 TI - [Neurophysiology and neurochemistry of the central processes related to pain and analgesia]. PMID- 3029202 TI - Extraction and delivery of the bone morphogenic factor as a substitute for hydroxylapatite in osseous augmentation. PMID- 3029203 TI - The use of sodium bicarbonate and hydrogen peroxide in periodontal therapy: a review. AB - The comparative benefits from the use of sodium bicarbonate and hydrogen peroxide over the use of a commercial dentifrice in periodontal therapy is controversial. The consensus of the clinical research indicates that application by patients of sodium bicarbonate and hydrogen peroxide offers no advantage over the preestablished, properly performed home oral hygiene procedures. Any improvements in clinical and microbial parameters generally were attributed to scaling and root planing. The studies that have reported beneficial results with sodium bicarbonate and hydrogen peroxide have used additional antimicrobial agents, concomitant professional application of these substances, and scaling and root planing. In one of these reports, inorganic salts and chloramine-T were delivered subgingivally throughout root-planing procedures, in addition to home application of inorganic salts. Most of these patients also received at least one course of systemic tetracycline therapy. Because this study had no control group, it is impossible to determine whether this program is more effective than are other periodontal therapy programs. A more controlled clinical study involving professional application of sodium bicarbonate, sodium chloride, hydrogen peroxide, and povidone-iodine has shown greater gains in clinical attachment and bone mass than has brushing with toothpaste and water. Again, subgingival scaling and root planing were necessary to attain these results. Because multiple topical agents were applied in both of these reports and systemic antimicrobial agents were used by the Keyes group, it is impossible to determine which agent was responsible for the improvements. Further, professional application may be the crucial factor.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3029204 TI - The injurious effect of eosinophil peroxidase, hydrogen peroxide, and halides on pneumocytes in vitro. AB - Recent data suggest that eosinophils may cause lung injury. To determine if the eosinophil peroxidase (EPO)-hydrogen peroxide (H2O2)-halide system could mediate this injury, we added human EPO, H2O2 (or glucose and glucose oxidase as a continuous source of H2O2), and various halides to monolayers of 51Cr-labeled human A549 and rat type II pneumocytes. Cell lysis was measured as soluble 51Cr release. In initial experiments, EPO in solution did not induce lysis under these conditions. Therefore, in subsequent experiments, pneumocytes were preincubated with EPO for 15 minutes, washed to remove unbound enzyme, and then glucose, glucose oxidase, and the halides were added. EPO alone was not injurious, nor was the addition of glucose and glucose oxidase in the absence of EPO. In contrast, the combined addition of EPO, glucose, glucose oxidase, and chloride produced marked target-cell lysis. This effect was time and EPO dose dependent and was enhanced by the addition of iodide. Catalase and azide substantially inhibited the lysis produced by the EPO-H2O2-halide system, suggesting that EPO-catalyzed products of halide oxidation mediated this form of injury. Finally, the addition of eosinophil major basic protein at 10(-5) mol/L to EPO-coated pneumocytes incubated with glucose, glucose oxidase, and halides failed to enhance or inhibit lysis. We hypothesize that the EPO-H2O2-halide system may injure the lung in asthma and eosinophilic pulmonary syndromes. PMID- 3029205 TI - The use of angiotensin converting enzyme inhibitors in elderly patients with hypertension. PMID- 3029206 TI - A demonstration that O2- is a crucial intermediate in the high quantum yield luminescence of luminol. AB - The chemiluminescence of luminol, due to its reaction with alkaline H2O2, is inhibited by superoxide dismutase or by hydroxyl radical scavengers. Hematin markedly enhances this H2O2-induced luminescence of luminol and lessens, but does not eliminate, the sensitivity towards these inhibitors. Reaction mechanisms are proposed to account for these results. Since luminol luminescence depends upon a reaction between the luminol radical and O2-, and since the luminol radical can reduce dioxygen to O2-, superoxide dismutase-inhibitable luminol luminescence cannot be reliably used as a detector of O2- production. PMID- 3029207 TI - Superoxide in ocular inflammation: human and experimental uveitis. AB - A possible involvement of superoxide in the pathogenesis of uveal inflammation in man and experimental animals was investigated. Superoxide production by the leukocytes of Behcet patients was significantly higher in the attack phase than in the remission phase. Leukocyte superoxide generation was also enhanced in guinea pigs with S-antigen-induced experimental autoimmune uveoretinitis (EAU). If the animals were treated with superoxide dismutase (SOD) at the onset of EAU, aqueous humor cell count was significantly lower than that of control (i.e., without SOD treatment). Infiltration of the inflammatory cells in the anterior retina was markedly reduced in SOD-treated animals. A similar protective effect of SOD against tissue damage was also observed in a bovine serum albumin-induced passive Arthus type uveitis in rabbits. These results suggest that superoxide may play a role in causing tissue damage in animal models of ocular inflammation and possibly in Behcet disease. PMID- 3029208 TI - Stimuli-induced superoxide radical generation in vitro by human alveolar macrophages from smokers: modulation by N-acetylcysteine treatment in vivo. AB - Bronchoalveolar lavage (BAL) was performed on nine healthy nonsmoking subjects and on 11 healthy smokers; in the last mentioned group lavage was performed before and after eight weeks treatment with N-acetylcysteine (NAC; 200 mg t.i.d.). The BAL cells were cultured for 2 h or overnight. Adherent cells were examined for their capacity to generate superoxide radicals (determined by superoxide dismutase (SOD)-inhibitable cytochrome C-reduction) at stimulation with phorbol 12-myristate 13-acetate (PMA), serum-treated zymosan (STZ), the calcium ionophore A23187, or the chemotactic tripeptide formyl methionylleucylphenylalanine (FMLP). Cells from nonsmokers responded with a very low degree of O(2)-generation to any of the stimuli employed whether cultured for 2 h or overnight. Cells from smokers also responded with low O(2)-generation after 2 h of culture. However, cells from smokers cultured overnight responded with marked O(2)-generation to PMA and STZ but the responses to FMLP and A23187 were low. NAC-treatment of the smokers resulted in a reduced degree of both PMA- and STZ-induced O(2)-generation in five individuals. In two other subjects, PMA induced (but not STZ-induced) O(2)-generation was reduced. Two individuals showed increased O(2)-generation to PMA- and to STZ-stimulation after NAC-treatment. Mean values of O(2)-generation induced by A23187 or by FMLP were significantly reduced for cells harvested after NAC-treatment. Mean values for PMA-induced O(2) generation also tended to be reduced by the treatment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3029209 TI - Increased spontaneous chemiluminescence from liver homogenates and isolated hepatocytes upon inhibition of O2- and H2O2 utilization. AB - The intracellular steady-state concentrations of hydrogen peroxide or superoxide anion were increased by inhibiting either catalase, glutathione peroxidase, or superoxide dismutase activities. Catalase was inhibited with aminotriazole while glutathione peroxidase activity was blocked by eliminating reduced glutathione after addition of either iodoacetamide diethylmaleate or phorone. The concentration of aminotriazole that stimulated chemiluminescence in 50% (60 mM) was very similar to the Ki for catalase activity (70 mM). Cyanide, an inhibitor of both catalase and superoxide dismutase, stimulated chemiluminescence in 50% at a concentration (0.15 mM) which is much closer from the Ki for superoxide dismutase (0.25 mM) than from the Ki for catalase (15 microM). The superoxide dismutase inhibitor diethyldithiocarbamate also increased chemiluminescence six- to ten-fold. Depletion of reduced glutathione stimulated spontaneous chemiluminescence when its concentration decreased below 4.5 mumol X g liver-1. The results shown herein suggest that the changes in the intracellular steady state concentration occurring after inhibition of any antioxidant enzyme are responsible for the increased spontaneous chemiluminescence. Spontaneous chemiluminescence from intact cells may be used as a noninvasive method for monitoring intracellular free radical metabolism. PMID- 3029211 TI - Pulse radiolysis study of daunorubicin redox reactions: redox cycles or glycosidic cleavage? AB - Two aspects of daunorubicin reactivity were investigated by pulse radiolysis. The reactions of O2 and O2- with the semiquinone and the hydroquinone transients of daunorubicin were determined and their rate constants measured. Although O2- can reduce the drug and its semiquinone form, it is a more powerful oxidant towards the two reduced transients. The hydroquinone daunorubicin glycosidic cleavage in aqueous solution was studied. Three intermediates were seen and characterized by their absorption spectra, their formation and decay kinetics. The competition between these two main processes was evaluated in the conditions of pulse radiolysis. Even under low O2 partial pressures the redox cycles are much more rapid than the glycosidic cleavage and a relatively high O2- steady state is settled. Biological implications are discussed. PMID- 3029210 TI - Changes in synaptic transmission produced by hydrogen peroxide. AB - The effect of hydrogen peroxide (H2O2) on excitatory and inhibitory synaptic transmission was studied at the lobster neuromuscular junction. H2O2 produced a dose dependent decrease in the amplitude of the junction potential (Vejp). This decrease was due to changes in both presynaptic transmitter release and the postsynaptic response to the neurotransmitter. Observed presynaptic changes due to exposure to H2O2 were a decrease in the amount of transmitter released, that is, quantal content, as well as a decrease in the fast facilitation, that is, the amplitude increase of successive excitatory junction potentials at a rate of 3 Hz. To discern postsynaptic changes, glutamate, the putative excitatory neurotransmitter for this preparation was applied directly to the bathing medium in order to bypass the presynaptic release process. H2O2 produced a decreased response of the glutamate receptor/ionophore. The action of H2O2 was not selective to excitatory (glutamate-mediated) transmission because inhibitory (GABA-mediated) transmission was also depressed by H2O2. This effect was primarily presynaptic since H2O2 produced no change in the postsynaptic response to applied GABA. PMID- 3029212 TI - Transferrin-dependent lipid peroxidation. AB - The potential for iron bound to transferrin to be released and promote the peroxidation of phospholipid liposomes was investigated using ADP as a low molecular weight chelator and superoxide generated by the xanthine/xanthine oxidase system as the reducing agent. Lipid peroxidation in this system was dependent upon transferrin as the source of iron; increasing the transferrin concentration resulted in increased rates of lipid peroxidation. Increasing the xanthine oxidase activity also caused increased rates of peroxidation. Catalase stimulated rates of peroxidation at all xanthine oxidase activities tested. Conditions resulting in the most rapid release of iron from transferrin (low pH, high ADP) did not promote the greatest rates of lipid peroxidation, indicating that at neutral pH, rates of lipid peroxidation may be limited by the availability of iron. It is concluded that transferrin is not a likely source of iron for catalysis of deleterious biological oxidations such as lipid peroxidation in vivo. PMID- 3029213 TI - Cellular and subcellular distribution of laminin in adult rat anterior pituitary. AB - Following the immunodetection of laminin in basal laminae and within some glandular cells of adult rat pituitary (Tougard et al., 1985: In Vitro 21:57), the present work had a double purpose: (a) adjustment of optimal conditions to detect laminin at both the cellular and subcellular level with an immunoperoxidase pre-embedding procedure, and (b) identification of the hormonal specificity of the laminin-containing cells, based on development of a "two-step" method combining the pre- and post-embedding approaches. In addition to extracellular localizations (basal laminae and connective tissue compartment), laminin was detected intracellularly within membrane-bounded compartments in all endocrine cell types and in the folliculo-stellate cells. It was found in rough endoplasmic reticulum cisternae, in a few Golgi saccules, and in vesicles or vacuoles. Labeled secretory granules were observed in gonadotrope, thyrotrope, and corticotrope cells but were very scarce in prolactin cells and absent in somatotrope cells. In addition, exocytotic profiles of labeled granules were frequent, suggesting a granular pathway for laminin export. Labeled vesicles, heterogeneous in size and shape, were observed mostly in prolactin cells. They might represent either a vesicular pathway for laminin export or an endocytic route for membrane-bounded laminin. These results bring complementary data to the concept that epithelial cells elaborate their basement membranes (Pierce and Nakane, 1967: Lab Invest 17:499). Moreover, the subcellular distribution of laminin within organelles involved in the biosynthesis, transport, and even in the storage of secretory products suggests that laminin might be exported by glandular pituitary cells according to different pathways. PMID- 3029215 TI - Quantitative assessment of Na+,K+-ATPase localization by direct and indirect p nitrophenyl phosphatase methods. AB - Na+,K+-ATPase histochemistry, using direct (Pb) and indirect (Mg/Sr-Co) p nitrophenyl phosphatase methods, was assessed by scanning integrative microdensitometry of three classes of tubules in mouse and guinea pig kidneys. The methods yielded similar and appropriate patterns of activity distribution and inhibitor response. The indirect method gave preferable results, in that non enzymic background was lower and rate of reaction product accumulation was considerably higher. PMID- 3029216 TI - Fat induced hypertension in rabbits: the effects of dietary linoleic and linolenic acid. AB - It has been suggested that dietary linoleic and gamma-linolenic acid may be hypotensive. Fat enriched diets increase blood pressure, and the effects of linoleic and linolenic acids on this increase have been investigated. After a control period on a low fat diet, rabbits were given four fat enriched diets containing different proportions of linoleic, gamma-linolenic and alpha-linolenic acids for 8 weeks, and returned to the low fat control diet for 3 weeks. Blood pressures were measured non-invasively every day. Blood pressures increased from the 4th week of fat feeding. The blood pressure increase at 8 weeks was 10%, 13%, 15% and 14% respectively for primrose, starflower, safflower and olive oils (all P less than 0.001). Return to the low fat control diet for 3 weeks restored blood pressures to near control values. These results do not support the suggestion that either linoleic or gamma-linolenic acids are effective antihypertensive agents. PMID- 3029214 TI - Simultaneous light and electron microscopic observation of immunolabeled liver 27 KD gap junction protein on ultra-thin cryosections. AB - We report on immunolabeling of gap junction protein in rat liver. Simultaneous light and electron microscopic immunolabeling of ultra-thin frozen sections was performed to confirm that the antigenic targets of polyclonal antibodies and a monoclonal 27 KD antibody (12/1 C5) are the gap junctions. Our results clearly demonstrate that the immunoreactive sites determined by indirect immunofluorescence correspond to immunogold-labeled gap junctions identified in the same section according to electron microscopic criteria. Our results also support the concept that the 27 KD protein is a major constituent of gap junctions. PMID- 3029217 TI - Central alpha-adrenoceptors during the development of hypertension in rats on high and low salt intake. AB - Our purpose was to investigate the binding characteristics of central alpha adrenoceptors during the early stages of the development of hypertension in rats on high and low salt (NaCl) intake. We measured alpha 1-[( 3H]prazosin) and alpha 2-[( 3H]rauwolscine) binding in membranes of the hypothalamus and medulla oblongata of six groups of young Wistar-Kyoto (WKY) rats, spontaneously hypertensive rats (SHR) and subtotally nephrectomized WKY (SN) rats with mean arterial blood pressure (MAP) ranging from normotensive to hypertensive levels after 1 week of salt restriction or loading. In the hypothalamus the SN-high salt rats and both SHR groups had elevated alpha 1-number but there was no change in alpha 2-number. Moreover, MAP was positively correlated with mean hypothalamic alpha 1-number in the six groups. In the medulla oblongata alpha 1-number was unaffected. However, high salt diet influenced medullary alpha 2-binding in the opposite manner in WKY rats versus SN rats and SHR. In these latter groups the affinity was increased and the number decreased in response to high salt intake. Furthermore, a positive correlation between MAP and mean alpha 1:alpha 2 ratio existed in both the hypothalamus and the medulla of the six groups. The data suggest that hypothalamic alpha 1-binding capacity was increased in SHR due principally to a genetic condition which is mimicked by salt loading in the SN rats. Medullary alpha 2-adrenoceptors of WKY, which remained normotensive despite salt loading, responded differently to high salt intake than those of the SN and SHR, whose blood pressure rose significantly.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3029219 TI - Angiotensin converting enzyme (ACE), characterization by 125I-MK351A binding studies of plasma and tissue ACE during variation of salt status in the rat. AB - Angiotensin converting enzyme (ACE) was measured in rat plasma and tissues by analysis of the binding of the radio-inhibitor 125I-MK351A, a tyrosyl derivative of enalaprilic acid. Labelled 125I-MK351A bound to plasma and tissue ACE preparations and was displaced in a concentration-related manner by MK351A. Scatchard analysis yielded a single line from which MK351A binding sites/mg protein and MK351A dissociation constant were calculated. Tissue and plasma ACE from Sprague Dawley rats was studied over a wide range of sodium intakes (low salt, 0.026 +/- 0.012 mmol/24 h; normal salt, 0.58 +/- 0.09 mmol/24 h; and high salt, 10.4 +/- 1.3 mmol/24 h) and during high salt and DOCA treatment. Across the range of sodium states studied there were no consistent changes in plasma, lung, aorta, brain, epididymis or kidney MK351A binding sites/mg protein, or equilibrium dissociation constant. Calculated MK351A binding sites (nmol/mg protein) were 1.64 +/- 0.14 in lung, 0.47 +/- 0.04 in aorta, 0.44 +/- 0.05 in epididymis, 0.18 +/- 0.01 in brain and 0.053 +/- 0.004 in kidney preparations (n = 24 in each group) reflecting reported ACE enzymatic activity in these tissues. Equilibrium dissociation constant KD was uniform within each tissue, but varied between organs. The KD (mol/l X 10(-12)) was 50 +/- 2 in aorta, 57 +/- 2 in lung, 58 +/- 2 in epididymis, 61 +/- 3 in brain, 62 +/- 2 in plasma and 84 +/- 3 in kidney (n = 24 in each group).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3029218 TI - Mechanism of action of a new prostaglandin antihypertensive, viprostol [CL 115 347; (dl)-15-deoxy-16-hydroxy-16(alpha/beta)-vinyl-prostaglandin E2 methyl ester]: (I). Vasodilation. AB - Viprostol [(dl)-15-deoxy-16-hydroxy-16(alpha/beta)-vinyl-prostaglandin E2 methyl ester; CL 115 347], injected directly into coronary, renal, mesenteric, femoral and carotid arteries of the anaesthetized beagle dogs at doses which did not lower the systemic arterial blood pressure, increased blood flow of the vascular beds being studied. Viprostol was as potent as I-prostaglandin E2 (I-PGE2) in the renal bed, but less potent in the other vascular beds. Viprostol was as potent as I-PGE2 in relaxing smooth muscle of the perfused isolated central ear artery of the rabbit. Viprostol maintained its antihypertensive effect in spontaneously hypertensive rats (SHR) pretreated with indomethacin, chlorpheniramine plus cimetidine, atropine or propranolol, suggesting that the vasodilating effects of viprostol were not mediated through the endogenous prostaglandin, histamine, cholinergic nervous system or beta-adrenergic nervous system. The antihypertensive effects of viprostol were not altered in SHR with bilateral nephrectomy or bilaterally ligated ureters, suggesting that the action of viprostol was not dependent on the secretory or excretory functions of the kidneys. In conclusion, viprostol exerts its antihypertensive action mainly by vasodilation in a way similar to the natural PGE2. PMID- 3029220 TI - Vitamin D affects proliferation of a murine T helper cell clone. AB - 1,25-Dihydroxyvitamin D3 (1,25(OH)2D3), the biologically active form of vitamin D3, has been shown to inhibit the activation of T cell hybridomas and heterogeneous populations of mononuclear leukocytes. Because the response of various clones to 1,25(OH)2D3 may differ, we have examined the proliferative effects of the steroid on an antigen-specific cloned, nontransformed T helper cell line (D10.G4.1 [D10 cells]), and find that in contrast to these previous studies, the steroid is a potent stimulator of lectin-induced proliferation. In these experiments, D10 cells were incubated with concanavalin A and 1,25(OH)2D3, and although the lectin or steroid alone has minimal proliferative effects, their co-addition prompts up to a 50-fold increase in 3H-TdR incorporation at a concentration of 2.5 to 5 X 10(-9) M 1,25(OH)2D3, with significant mitogenesis occurring at 0.1 to 0.3 X 10(-9) M 1,25(OH)2D3. 25-Hydroxyvitamin D3 and 24,25(OH)2D3 have similar activity, but at concentrations two to three times greater than that of 1,25(OH)2D3, reflecting their relative affinities for the 1,25(OH)2D3 receptor. In addition, lectin treatment enhances 1,25(OH)2D3 receptor capacity fourfold to fivefold, an event coupled with the appearance of positive cooperativity. Although the steroid does not affect the quantity of bioassayable T cell growth factors as assessed by HT-2 cell proliferation, the expression of immunoreactive IL 2 receptors by lectin-activated D10 cells exposed to 1,25(OH)2D3 is enhanced. In contrast to its proliferative effect in the absence of IL 1, 1,25(OH)2D3 exerts biphasic effects on D10 replication when this monokine is present. Specifically, this steroid augments D10 proliferation at low concentrations of recombinant IL 1, but as the abundance of the monokine increases in the presence of 10(-10) to 10(-8) M 1,25(OH)2D3, the peak response of D10 cells to optimal IL 1 concentrations is diminished. Therefore, in this clone, 1,25(OH)2D3 presents itself as a regulator of T helper cell proliferation. PMID- 3029221 TI - Immunoregulatory activity of recombinant human tumor necrosis factor (TNF)-alpha: induction of TNF receptors on human T cells and TNF-alpha-mediated enhancement of T cell responses. AB - The expression of specific tumor necrosis factor (TNF) membrane receptors and biological effects of recombinant TNF (rTNF)-alpha on normal human T lymphocytes were studied. Although resting T cells lacked specific binding capacity for rTNF alpha, high affinity (Kd 70 pM) TNF receptors were de novo induced upon primary activation of T cells. Comparison of TNF receptor expression with that of high affinity interleukin 2 (IL-2) and interferon-gamma (IFN-gamma) receptors, respectively, revealed similarities to IL 2-receptor expression with respect to kinetics of induction. However, maximum expression of TNF receptors (approximately equal to 5000/cell at day 6) and subsequent decline occurred approximately 3 days after the peak of IL 2-receptor expression. In contrast, no change in the expression of IFN-gamma receptors (Kd 10 pM, 300 to 400 receptors/cell) was found in the course of T cell activation. On activated TNF receptor positive T cells, TNF-alpha exerted multiple stimulatory activities. Thus TNF increased the expression of HLA-DR antigens and high affinity IL 2 receptors. As a consequence, TNF-treated T cells showed an enhanced proliferative response to IL 2. Moreover, TNF-alpha was effective as a co-stimulator of IL 2 dependent IFN-gamma production. These data indicate that TNF-alpha may regulate growth and functional activities of normal T cells. PMID- 3029222 TI - Antibody-independent activation of C1. I. Differences in the mechanism of C1 activation by nonimmune activators and by immune complexes: C1r-independent activation of C1s by cardiolipin vesicles. AB - C1 activation is controlled by the regulatory protein C1-inhibitor (C1-INH). In contrast to immune-complex-induced activation, which is insensitive to C1-INH, antibody-independent activation of C1 is modulated by C1-INH. The mechanisms regulating nonimmune activation were studied with two phospholipids varying in their capacity to activate C1 in the presence of C1-INH: cardiolipin (CL) and phosphatidylglycerol (PG). Whereas C1-INH consistently suppressed activation by PG vesicles, a dose-dependent increase in C1 activation was measured with CL vesicles above 40 mole %. A similar dose-response binding of C1s requiring C1q, but not C1r, was detected only on CL vesicles, but neither on PG vesicles nor on immune complexes. This binding was Ca2+-dependent, suggesting that dimeric C1s is involved and was inhibited by spermine. The C1q-bound C1s was specifically cleaved at 37 degrees C into its active 58 kDa and 28 kDa chains, in the absence of C1r. On the addition of anti-CL antibodies, the C1q-mediated cleavage of C1s by CL vesicles was specifically inhibited. The cleavage of C1r on CL vesicles was also determined. When macromolecular C1 was offered in the presence of C1-INH, C1r cleavage was detected; however, the presence of C1s was a critical factor for C1r activation, because it was required on CL vesicles, but not on immune complexes. These results show that nonimmune activation of C1 presents specific features which distinguish it from immune complex-induced activation. These characteristics varied with the capacity of antibody-independent activators to activate C1 in the presence of C1-INH. PMID- 3029223 TI - Antibody-independent activation of C1. II. Evidence for two classes of nonimmune activators of the classical pathway of complement. AB - Nonimmune activation of the first component of complement (C1) by cardiolipin (CL) vesicles present specific features which were not demonstrated on immune complexes. CL vesicles which activate C1 in the presence of C1-inhibitor (C1-INH) were found to bind C1s in the absence of C1r, and to induce a specific C1r independent cleavage of C1q-bound C1s. Therefore, several known natural nonimmune activators were analyzed by comparing their ability to activate C1 in the presence of C1-INH and to mediate a C1r-independent cleavage of C1s. Freshly isolated human heart mitochondria (HHM) activated C1 only in the absence of C1 INH. However, mitoplasts derived from HHM (HHMP) activated C1 regardless of the presence of C1-INH, and induced a specific cleavage of C1q-bound C1s. The same pattern was observed in the case of smooth E. coli and a semi-rough E. coli strain. DNA, known to activate C1 only in the absence of C1-INH, does not induce C1s cleavage in the absence of C1r. Thus, nonimmune activators can be classified into two distinct categories. "Strong" activators, such as CL vesicles, HHMP, or the semi-rough E. coli strain J5 can activate C1 in the presence of C1-INH. By using C1qs2 as a probe, they exhibit a specific, C1r-independent cleavage of C1s. C1s-binding to C1q is a critical factor for the activation process in this group. In the case of "weak" activators, such as E. coli smooth strains, DNA, or HHM, no C1s-binding to activator-bound C1q was detected, and C1r-independent C1s cleavage and C1 activation in the presence of C1-INH were not observed. As in the case of immune complexes, C1r activation appears to play a key role in the C1 activation by "weak" activators. PMID- 3029224 TI - Arachidonate metabolites, platelet-activating factor, and the mobilization of protein kinase C in human polymorphonuclear neutrophils. AB - In contrast to our previous report (Biochem. Biophys. Res. Comm. 134:587, 1986), we now find that protein kinase C (PKC) is mobilized in human polymorphonuclear neutrophils (PMN) stimulated with platelet-activating factor (PAF) or leukotriene (LT)B4. Thus nanomolar concentrations of each compound caused PMN to lose cytosolic, PKC-specific protein phosphorylating activity, as well as receptors for phorbol myristate acetate (PMA). Smaller gains in membrane-associated PMA receptors accompanied these changes. Diacylglycerol and PMA had very similar effects on PKC. However, unlike these direct PKC activators, PAF and LTB4 induced only moderate decreases in cytosolic PKC; acted only on PMN pretreated with cytochalasin B; did not mobilize PKC in disrupted PMN or activate PKC in a cell free system; and with respect to PAF, induced responses that partially reversed within 30 min. Furthermore, PAF, LTB4, and several of their structural analogues mobilized PKC at concentrations correlating closely with their respective affinities for cellular LTB4 or PAF receptors. Thus PAF and LTB4 acted by indirect and apparently receptor-mediated mechanisms. Four observations indicated that the cytochalasin B-dependent degranulating actions of PAF and LTB4 involved PKC. First, PKC mobilization and degranulation occurred at the same stimulus concentrations. Second, 5-hydroxyicosatetraenoate dramatically enhanced both PKC mobilization and degranulation when elicited by PAF; it had relatively little influence on LTB4-induced responses. Third, PAF-induced mobilization (t1/2 less than 7 sec) preceded degranulation (t1/2 approximately 20 sec). Finally, a PKC blocker, polymyxin B, was similarly effective in inhibiting degranulation responses to PAF, LTB4, and PMA. Because stimulated PMN may produce and use PAF, LTB4, and 5-hydroxyicosatetraenoate as secondary intracellular mediators, our results implicate PKC as a central and potentially critical regulator of function. PMID- 3029225 TI - The immunoglobulin heavy chain switch: structural features of gamma 1 recombinant switch regions. AB - The immunoglobulin heavy chain isotype switch is mediated by a DNA rearrangement involving specific genomic segments referred to as switch regions. Switch regions are composed of tandemly repeated simple sequences. The role of the tandemly repeated structure of switch regions in the switch recombination process is not understood. We mapped eight recombination sites--six in the gamma 1 and two in the gamma 3 tandem arrays. In addition, we obtained molecular clones representing three of the six gamma 1 rearrangements, and determined the nucleotide sequences of the recombination sites in each. In general, the rearrangements are confined to the tandem repeat units, and are not clustered in a particular portion of either the gamma 3 or gamma 1 switch region. Nucleotide sequence analysis of one of the recombinant clones, gamma M35, reveals evidence for a successive switch event wherein a recombination between S mu and S gamma 3 was followed by recombination 57 bp downstream with S gamma 1. gamma 1 sequence data from the molecular clones we obtained, together with similar data from other investigators regarding the gamma 1, gamma 2b, and gamma 2a switch regions, reveals that recombinations tend to occur at homologous positions of the respective gamma-unit repeats, adjacent to the elements AGCT and GGGG found in each. This finding suggests that the cutting and religation step of the recombination process is mediated by a recombinase common to the four gamma-isotypes. PMID- 3029226 TI - Use of a liquid cell culture assay to quantify the elimination of Burkitt lymphoma cells from the bone marrow. AB - A new liquid cell culture assay for propagating Burkitt lymphoma (BL) cells in the presence of excess allogeneic irradiated bone marrow (BM) has been standardized. The Burkitt cell lines used in this assay were newly established from the tumours of our patients and were either EBV(+) or EBV(-). The phenotypic characteristics of lines were similar to those of the original malignant tumours. With the help of this liquid assay it was possible to evaluate the efficacy of in vitro procedures designed to completely eliminate malignant cells from harvested BM prior to autologous transplantation (i.e., immunomagnetic depletion or antibody-complement-mediated cytolysis). With six out of the seven cell lines tested, the assay permitted the detection of as few as one BL cell in 4 X 10(6) normal BM cells and the measurement of up to 5 log BL cell elimination from 1% contaminated samples. PMID- 3029227 TI - A specific assay measuring binding of 125I-Gp 120 from HIV to T4+/CD4+ cells. AB - The HIV (HTLV-III) envelope glycoprotein, Gp120, was isolated from virus-infected tissue culture cells using affinity chromatography. A radioimmunoassay was developed to determine the degree of iodinated Gp120 to target CD4+ (T4+) cells. 125I-Gp120 could be shown to selectively bind to CD4+ cells only. The Gp120 remained bound to these cells after repeated washes. Monoclonal anti-CD4 antibodies block the binding of Gp120 to CD4+ cells. Monoclonal antibodies to other cell surface components do not interfere with 125I-Gp120 binding. All IgG antibodies from HIV seropositive donors tested block 125I-Gp120 binding, though with variable titers. We believe that this assay provides further proof for the use of CD4 (T4) as a component of the receptor for HIV. It represents a safe, objective and sensitive method for the analysis of Gp120-CD4 interactions, as well as the potential of antibodies to interfere with this binding. PMID- 3029229 TI - Lucigenin chemiluminescence in the assessment of neutrophil superoxide production. AB - Lucigenin-amplified chemiluminescence (LUCL) was induced by .O2- (from the xanthine/xanthine-oxidase reaction) but not, as with luminol-augmented CL (LCL), by myeloperoxidase (MPO) and only weakly by H2O2 or the H2O2-MPO-Cl- system. Neutrophil LUCL induced by fMet-Leu-Phe (fMLP) was dose-dependently inhibited by scavengers of .O2- but not by NaN3, catalase, mannitol or taurine. Response patterns of several soluble stimuli (leukotriene B4 (LTB4), platelet-activating factor (PAF), fMLP, A23187 and phorbol-myristate-acetate (PMA)) were assessed with LUCL. PMA-induced LUCL was protracted, A23187 showed intermediary kinetics whereas fMLP and PAF both showed rapid peak responses at 30-50 s. However, LTB4 was the most rapid initiator of LUCL (7-12 s). The kinetics of fMLP- or PMA induced neutrophil .O2- production, assessed with the cytochrome C reduction method, correlated well with LUCL. Thus it is suggested that LUCL provides a specific method for studying the kinetic properties of neutrophil .O2- production where stimulus-specific response patterns can be distinguished. PMID- 3029228 TI - Preparation and characterization of mast cell cytoplasts. AB - A new model system for studying biochemical reactions in mast cell plasma membranes was developed. Particles termed cytoplasts consisting of organelle depleted cytoplasm surrounded by an intact plasma membrane were formed from cytochalasin B-treated mast cells ultracentrifuged through a discontinuous Ficoll gradient. Two cytoplasts were formed per mast cell and 95% were recovered. Mast cell cytoplasts had a mean diameter of 3.2 microns with a median volume of 38 microns 3. Enzyme marker studies indicated that subcellular recoveries in the mast cell cytoplast were: plasma membrane = 16%, cytoplasm = 39%, nucleus = 1.1%, granule = 0.5%. Analysis of IgE receptors indicated that mast cell cytoplasts retained the normal asymmetric orientation of the plasma membrane. Mast cell cytoplasts synthesized ATP, incorporated labeled fatty acids into complex lipids and retained fluorescein after deacylation of diacetylfluorescein. The quantity of cAMP (adenosine 3':5'-cyclic monophosphate) maintained in mast cell cytoplasts was 0.0304 pmol/10(6) original mast cells. Cytoplasts offer the opportunity to study plasma membrane and cytoplasmic biochemical events that occur during stimulation in a relatively physiologic environment. PMID- 3029230 TI - The effects of adherent cells on measurement of the hyper-responsiveness of rheumatoid B lymphocytes to Epstein-Barr virus. AB - Rheumatoid peripheral blood mononuclear cells show an increased responsiveness to superinfection with Epstein-Barr virus (EBV). We have investigated the role of adherent cells in this hyperresponsiveness using two different methods of adherent cell depletion. Depletion of adherent cells from both rheumatoid and normal mononuclear cells, using either dextran bead columns or plastic petri dishes, produced inconsistent changes in the response of autologous non-adherent cells to EBV. The addition of supernatants of cultured rheumatoid adherent cells also produced an inconsistent change in response although normal adherent cell supernatants increased the responsiveness of autologous non-adherent cells to EBV. The inconsistencies observed are discussed with respect to adherent cell subpopulations present in rheumatoid and normal peripheral blood mononuclear cell preparations when using recognized methods of adherent cell depletion. PMID- 3029231 TI - Aberrant MHC antigens in a sarcoma virus-induced mouse tumour. AB - From a series of mouse sarcomata, newly induced by Rous sarcoma virus (RSV), the DOH cell line was shown to lack expression of syngeneic H-2Kd and Dk antigens (Noll et al., 1986). It unexpectedly displayed determinants specific for H-2Kk molecules. Interferon treatment of DOH stimulated the expression of H-2Kk determinants and also the display of some, but not all, determinants of the syngeneic H-2Kd molecules. H-2Dk expression was not stimulated. Southern blot hybridization of genomic DNA digests from DOH cells confirmed the identity of the H-2K region with that from syngeneic C3H.OH liver cells, but also showed changes in the pattern of restriction fragments that contain class I genes from the D and Qa regions. These results suggest that aberrant MHC class I molecules that carry H-2Kd- and H-2Kk-like determinants are expressed by DOH sarcoma cells. These molecules may act as target antigens for tumour-specific cytotoxic T cells elicited by injection of DOH cells into syngeneic mice. PMID- 3029232 TI - Heterogeneity of the expression of class I and II HLA antigens in human breast carcinoma. AB - HLA class I and II antigen expression was studied in 19 cases of primary infiltrating ductal carcinoma of the breast. An indirect immunofluorescence technique was used on cryostat sections with monoclonal antibodies directed against HLA class I and II monomorphic determinants. Of the 19 cases studied, 17 were positive for class I antigen expression and two were negative. Class I HLA antigen expression was found to be clearly heterogeneous: in ten of these tumours more than 75% of the cells were class I positive; in two the percentage was decreased to between 50% and 75%; in five tumours it was less than 50%. With respect to class II HLA antigen expression, eight breast tumours were totally negative while two were strongly positive (50-75%) and the nine remaining cases were less than 25% positive. In addition, radioassay techniques were employed to determine the presence of oestrogen and progestagen receptors. The distribution of these receptors was not correlated with HLA class I or II antigen expression, nor could any relationship be demonstrated between the degree of histological differentiation of the tumours and their invasiveness. PMID- 3029233 TI - Different sensitivity of B cell lines derived from Burkitt lymphoma (BL) and normal cells (LCL) to cytotoxic T cell clones generated by autologous and allogeneic stimulation. PMID- 3029234 TI - The H-2 complex regulates both the susceptibility to mouse viral lymphomagenesis and the phenotype of the virus-induced lymphomas. AB - Neonatal infection of C57BL/10 mice with cloned ecotropic and/or dualtropic mink cell focus-inducing (MCF) mouse leukaemia viruses (MuLV), induces a wide spectrum of different lymphomas of T, B, and non-T/non-B cell types. The H-2 complex has a marked influence on both the development of lymphoma incidence and lymphoma type. A study using the oncogenic MCF 1233 virus and a series of B10 congenic mice enabled the mapping of the following: Resistance to the early development of T cell lymphoma is controlled by the H-2I-A locus. Susceptibility to early T cell lymphomagenesis is associated with an I-A-regulated low anti-MCF 1233 envelope antibody response and persistent infection of the thymus. B10 (H-2b) mice, which are resistant to early T cell lymphomagenesis induced by MCF 1233 or other MuLV isolates, have high anti-MuLV envelope antibody responses which are I-A regulated. These mice develop more B cell lymphomas late in life in contrast to the early development of T cell lymphoma in B10.A (H-2a) mice. The possible response mechanisms which underlie these observations, including: I-A-regulated immunoselection against MuLV antigens expressed by (pre) leukaemic T cells, aberrant expression of class II MHC antigens on some B cell lymphomas and I-A regulated chronic immunostimulation of MuLV-expressing (pre) leukaemic B cells, are discussed. PMID- 3029235 TI - Infection with human parvovirus (B19), aplasia of the bone marrow and a rash in hereditary spherocytosis. AB - Infection with human parvovirus (HPV) B19 was diagnosed in persons with hereditary spherocytosis during an outbreak of erythema infectiosum. A 10-year old boy had an aplastic crisis of the bone marrow and a maculopapular rash. His mother also had an aplastic crisis, fever, rash and a transiently acellular marrow. Another boy had an aplastic crisis, leucopenia and fever, but did not have a rash. Aplastic crisis and a rash have rarely before been observed together and attributed to infection with HPV. PMID- 3029237 TI - Desmoplastic basal cell carcinomas possess unique basement membrane-degrading properties. AB - Desmoplastic basal cell carcinomas (fibrosing or morphea types) were studied ultrastructurally, immunocytochemically, and biochemically for basement membrane degrading activity and compared with the common varieties of basal cell (superficial and nodular-ulcerative types). Whereas the latter lesions demonstrated intact basement membranes as evidenced by extracellular laminin and type IV collagen immunoreactivity and the presence of an unusually thickened basal lamina, desmoplastic basal cell carcinomas showed large defects and absences in basal lamina and basement membrane immunoreactivity. Intense tumor cytoplasmic immunoreactivity for type IV collagenase was present in 13 of 15 cases of desmoplastic basal cell but absent in the superficial and nodular ulcerative varieties. Whereas explant cultures of all the types of basal cell carcinoma studied gave rise to high levels of interstitial (type I) collagenase activity in conditioned media, only the desmoplastic variety exhibited high type IV collagenase activity. These findings suggest that the mechanisms by which the desmoplastic and the common varieties of basal cell carcinoma infiltrate host tissues may be fundamentally different. PMID- 3029236 TI - Localization of the alpha 3 (V) chain of type V collagen in human skin. AB - Serum from goats immunized with human type V collagen chains that were cut out of polyacrylamide gels contained an antibody that recognized only type V collagen in an enzyme-linked immunoabsorbent assay and did not label laminin, fibronectin, or types I and IV collagen. Western blot analysis of the antibody showed that its determinant was the alpha 3 (V) chain of type V collagen. Indirect immunofluorescent staining of intact human skin with the antibody produced staining of the dermal blood vessels but not of the dermal-epidermal junction (DEJ). In contrast, both the dermal blood vessels and the DEJ were labeled by the antibody if the skin substrate was first split through the lamina lucida region of the DEJ by incubation in 1 M NaCl solution. Indirect immunoelectron microscopy confirmed the staining pattern found by immunofluorescence and defined the ultrastructural localization of type V collagen in skin. Type V collagen is localized within the DEJ to the lamina lucida region and polar aspects of the basal cell keratinocyte plasma membrane. PMID- 3029238 TI - Detection of human T cell leukemia virus type I and human immunodeficiency virus in cultured lymphocytes of a Zairian man with AIDS. AB - We report the detection of human T cell leukemia virus type I (HTLV-I) and human immunodeficiency virus (HIV) in the cultured lymphocytes of a 45-year-old Zairian man with AIDS. HIV was successfully isolated and analyzed by SDS-PAGE and competition radioimmunoassay. However, by the culture techniques used, HTLV-I could not be separated from the HIV. Western blot analysis of the patient's serum showed the presence of both HTLV-I- and HIV-specific antibodies. The finding of this dual infection may explain reports that greater than or equal to 30% of patients with AIDS are positive for antibodies to HTLV-I. PMID- 3029239 TI - Molecular epidemiology of live, attenuated varicella virus vaccine in children with leukemia and in normal adults. AB - Restriction endonuclease analysis of varicella-zoster virus (VZV) DNA has been used in unraveling the complex epidemiology of VZV infections in individuals immunized with a live, attenuated varicella virus vaccine. Early rashes appearing within the first six weeks after vaccination are invariably due to vaccine virus. True breakthrough infections with wild-type VZV also occur in vaccinees. Five cases of zoster have been seen in leukemic children vaccinated while in remission. One case appeared 22 months after vaccination in the same general area as the inoculation. The virus isolated was vaccine derived. A second case of zoster appeared in a dermatome unrelated to the sites of vaccination approximately 19 months after apparently natural varicella. This virus was wild type. Vaccine virus can therefore establish latency and can later reactivate as herpes zoster. PMID- 3029240 TI - Effect of B cell suppression on primary infection and reinfection of mice with herpes simplex virus. AB - During primary infection with herpes simplex virus type 1, elimination of the antibody response by B cell suppression did not interfere with clearance of virus from skin. However, spread of virus within the peripheral nervous system was more extensive in B cell-suppressed animals compared with that in fully immunocompetent mice. Mice that had previously recovered from herpes simplex virus infection of the ear pinna, when subsequently reinfected in the flank, showed restricted viral replication in the skin and cleared virus more rapidly than did animals experiencing primary infection. B cell suppression did not compromise the ability of mice to resist reinfection compared with the ability of normal immune animals. The implications of the above findings with regard to future immunoprophylaxis and the interpretation of experiments designed to test for immunity are discussed. PMID- 3029241 TI - Isolation of multiple strains of cytomegalovirus from women attending a clinic for sexually transmitted disease. AB - To study the occurrence of exogenous reinfection with cytomegalovirus (CMV), we examined serial isolates from the cervix, urine, and throat of eight women attending a clinic for sexually transmitted diseases (STD) and seven women receiving routine prenatal care. Isolates were compared by restriction enzyme "fingerprinting" of viral DNA. Four of the eight women from the STD clinic were infected with more than one strain of CMV. Two women shed different strains in serial isolates, and two women shed different strains simultaneously from different body sites. These results suggest that exogenous reinfection is not uncommon in women with a high probability of exposure to CMV. PMID- 3029242 TI - Early-stage developmental abnormalities induced by murine cytomegalovirus. AB - The onset of the effects of murine cytomegalovirus (MCMV) infection on an early stage of embryonic development (nine days) in a mouse model was studied with the aid of scanning electron microscopy. MCMV was injected into the endometrial lumina of pregnant mice at the time of implantation. The mice were later killed, and sites of embryonic implantation were examined. Compared with uninfected mice and mice inoculated with heat-inactivated virus, litter sizes were reduced, and the incidence of abnormal fetuses was significantly increased among MCMV-infected animals. Scanning electron microscopy also revealed maldeveloped cranial regions characterized by an unclosed neural tube and severely underdeveloped head. Ectodermal abnormalities, including poxlike formations and ballooning cells, were observed in several embryos. Thus, early cytomegalovirus infection may not only result in fetal loss, but may also interfere with the process of morphogenesis. PMID- 3029243 TI - Protection from endotoxemia: a rat model of plasmapheresis and specific adsorption with polymyxin B. AB - Polymyxin B (PB), a cyclic polypeptide antibiotic, has potent antiendotoxin properties but is associated with significant toxicity when given parenterally. As an alternative, PB was immobilized on a solid phase (Sepharose 4B; Pharmacia, Uppsala, Sweden), and a system of plasmapheresis was developed in the conscious rat, with specific on-line plasma adsorption of endotoxin by a PB-Sepharose column. PB-Sepharose columns removed 94% of a challenge dose of 5 micrograms of endotoxin in vitro. Rats were pretreated with lead acetate so that they were sensitized to endotoxin and then given 10 micrograms of endotoxin/kg intraarterially. After 15 min plasmapheresis was begun and continued for 90 min. Animals whose plasma was perfused over PB-Sepharose were protected from endotoxin induced leukopenia (P less than .01), thrombocytopenia (P less than .001), and death (four of four survivors compared with none of four controls). Thus plasmapheresis with on-line removal of endotoxin is a safe and highly effective means of protecting animals from the effects of endotoxemia. PMID- 3029244 TI - Levels of interferon in the plasma and cerebrospinal fluid of patients with acute Japanese encephalitis. PMID- 3029246 TI - [Primary malignant cardiac tumors: a report of four cases]. PMID- 3029245 TI - Acalculous cholecystitis and cytomegalovirus infection in a patient with AIDS. PMID- 3029247 TI - [Inhibition of human ovarian cancer cell growth by prostaglandin D2]. AB - The effects of prostaglandin D2 (PGD2) on human ovarian tumor growth were examined in vitro and in vivo by using a cell line, designated HR, derived from patients with serous cystadenocarcinoma of the ovary. The cell proliferation was dose-dependently inhibited by PGD2 between concentrations of 0.1 and 4.0 micrograms/ml after 24 hrs, 48 hrs and 72 hrs of contact time. Concentrations of PGD2 required for 50% inhibition of the cell proliferation were 2.0, 1.1 and 0.55 micrograms/ml with 24, 48 and 72 hrs of contact time, respectively. From the results of 51Cr-release assay, the inhibition of cell proliferation by PGD2 was considered to result from the direct cytotoxic effects. The incorporations of 3H thymidine, 3H-uridine and 3H-valine were inhibited in a dose-dependent fashion with more than 1.0 micrograms/ml of PGD2. Tumor growth in nude mice treated with 0.3 mg/mouse PGD2 was significantly inhibited, compared to that of untreated nude mice. In untreated nude mice the tumor growth curve was parallel to the changes in the plasma alpha-hydroxybutyrate dehydrogenase (HBD). In both PGD2-treated groups with 0.1 mg/mouse and 0.3 mg/mouse, the HBD activity markedly decreased on the 14th and the 21st day after inoculation. The 50% survival time in untreated mouse, 0.1 mg/mouse and 0.3 mg/mouse PGD2-treated groups was 52 days, 55 days and 67 days, each respectively. PMID- 3029248 TI - [Mixed mullerian tumor]. PMID- 3029249 TI - Separation of Mycobacterium leprae from contamination with armadillo-liver derived "pigment" particles. AB - Mycobacterium leprae isolated from armadillo liver by the widely used IMMLEP protocol is sometimes contaminated with a particulate "pigment." This paper describes a simple, efficient, and rapid method for purifying large quantities of contaminated bacteria, which may readily be used as an additional step added at the end of the protocol when necessary. The process involves a discontinuous Percoll gradient and generates an essentially pure fraction containing greater than 90% of the original bacteria, and a fraction of "pigment" slightly contaminated with bacteria. Use of the system should release large additional numbers of pure M. leprae suitable for use in human vaccine trials. PMID- 3029250 TI - [Hydroxyapatite-binding phosphoproteins synthesized by osteoblasts and fibroblasts]. PMID- 3029252 TI - [Myelotoxicity due to cancer chemotherapy]. PMID- 3029251 TI - [Evaluation of the combined procedure of Hp-D (hematoporphyrin derivative) phototherapy and radiation therapy in advanced lung cancer]. PMID- 3029253 TI - [Cell kinetic studies of the effects of cis-diamminedichloro platinum (II) on lung cancer cell lines]. PMID- 3029254 TI - [alpha 1-Antitrypsin deficiency with pulmonary emphysema, small cell lung cancer and marked elevation of serum lactic dehydrogenase level: a case report]. PMID- 3029255 TI - The neurohypophysis: recent developments. AB - The hormones of the neurohypophysis, vasopressin and oxytocin, have now been shown to be synthesized as part of a prohormone complex that includes a vasopressin-neurophysin and oxytocin-neurophysin, respectively. In addition, for vasopressin, there is a glycopeptide as part of the prohormone. For each hormone the prohormone is packaged into neurosecretory granules and transported via axons to the posterior pituitary gland. In addition to this "classic" system, axons containing neurohypophyseal hormones project to the median eminence for release into portal vessels, and to other areas of the brain and spinal cord where the peptides may function as neurotransmitters rather than as hormones. As neurotransmitters, the neurohypophyseal hormones may be involved in the regulation of certain autonomic functions. Vasopressin and oxytocin are secreted into the cerebrospinal fluid where there is a diurnal rhythmic secretion of the peptides in several animal species (some species have a predominant rhythm of vasopressin and others a rhythm of oxytocin). Neurohypophyseal peptides are synthesized in some non-neuronal tissues where the function is unknown, and recently a novel peptide with similarities to oxytocin and vasotocin has been identified. The relationship of this novel peptide to the neurohypophyseal peptides is unknown. These new developments are likely to elucidate many new functions for the hormones of the neurohypophysis. PMID- 3029256 TI - Triiodothyronine-induced cyst formation in metanephric organ culture: the role of increased Na-K-adenosine triphosphatase activity. AB - Previous organ culture investigations into the pathogenesis of renal cyst formation have demonstrated that glucocorticoid-induced proximal tubular cyst formation is associated with increases in renal sodium-potassium adenosine triphosphatase (Na-K-ATPase) activity. To explore the relationship between cyst production and transport enzyme induction, we examined the effects of the potent inducer of Na-K-ATPase activity, L-3,5,3'-triiodothyronine (T3), on renal tubular morphologic and enzymatic development in murine metanephric organ culture. The addition of T3 (2 X 10(-8) mol/L) to completely characterized, serum-free growth medium produced striking proximal tubular cystic abnormalities. Frank cyst development was preceded by ultrastructural alterations consisting of basolateral intercellular spreading, which increased with progressive tubular dilation. Ultrastructural analysis demonstrated no abnormalities of tubular cyst wall basal laminae, and immunohistologic staining with affinity-purified antibodies to the basal lamina glycoproteins fibronectin, laminin, and entactin, revealed no differences between cystic and control tissue. With use of an enzyme-linked kinetic microassay, T3-induced cystic organ culture explants (CY) showed significant increases in Na-K-ATPase when compared with controls from 72 to 120 hours of organ culture incubation. The initial differences in CY Na-K-ATPase occurred contemporaneously with the earliest ultrastructural evidence of cyst formation, and subsequent increases paralleled progressive tubular cyst formation. Tubular cyst formation in CY could be largely prevented by daily incubation of explants with ouabain, 0.2 mmol/L (final concentration) for 120 minutes without deleterious effects on overall metanephric development. We conclude that T3 induces proximal tubular cyst formation in metanephric organ culture, and that T3-induced increases in Na-K-ATPase have a primary role in the pathogenesis of tubular cyst formation in this model system. PMID- 3029257 TI - Oncogenes from hormone receptors. PMID- 3029258 TI - Effect of adrenaline on basal and ovine corticotrophin-releasing factor stimulated ACTH secretion in man. AB - Six normal male subjects were given, in single blind random order on six separate occasions, i.v. bolus doses of synthetic ovine corticotrophin-releasing factor-41 (oCRF-41; 25 and 50 micrograms) with and without adrenaline (3 micrograms/min) i.v. for 150 min, the adrenaline infusions alone and saline placebo. The adrenaline infusions resulted in plasma adrenaline concentrations of 4.33 +/- 0.82 (S.E.M.) nmol/l and were associated with an increase in blood glucose, heart rate and systolic blood pressure and a reduction of diastolic blood pressure. Despite these evident biological effects at several sites, there was no stimulation of plasma ACTH or cortisol by adrenaline in comparison with the effect of saline, and no enhancement of the stimulatory effect of either dose of oCRF-41 on ACTH or cortisol secretion. The ACTH response to 50 micrograms oCRF-41 was greater than that to 25 micrograms, indicating that the 25 micrograms dose of oCRF-41 was submaximal and capable of further enhancement. As the plasma adrenaline concentrations during the adrenaline infusions reached the upper limit of the physiological range of plasma adrenaline in man, yet failed to enhance the ACTH or cortisol responses to a submaximal dose of oCRF-41, we conclude that circulating adrenaline neither exerts a direct stimulatory effect on pituitary corticotrophs nor enhances the effect of CRF under physiological circumstances. The adrenaline infusions attenuated the ACTH and cortisol responses to oCRF-41 and were associated with a transient reduction of basal concentrations of both hormones. PMID- 3029259 TI - Enhanced corticosterone-binding capacity in plasma after stimulation of the rat adrenal cortex: the possibility of a simultaneous release of protein and corticosterone. AB - Exposure of rats to either footshock or handling stress produced a significant increase in both plasma corticosterone concentration and specific binding capacity. Non-specific binding was eliminated using the synthetic glucocorticoid, dexamethasone. The increase in both plasma corticosterone and specific binding capacity was biphasic following exposure to footshock. Adrenalectomy and pretreatment with betamethasone abolished both phases of the enhanced binding capacity and plasma steroid concentration. Intraperitoneal injection of ACTH (1 24) in animals pretreated with betamethasone resulted in a biphasic rise in plasma concentrations of corticosterone but only the initial increase in binding capacity. Dissociation constant (Kd) values, determined by Scatchard analysis, for adrenalectomized and betamethasone-pretreated animals were 546 and 556 pmol/l respectively. These values were significantly different from the Kd in animals with functional adrenals (631 pmol/l). The results are discussed in the light of a possible specific corticosteroid-binding globulin (CBG) like binding protein of adrenal origin released in conjunction with corticosterone. This binding protein has a lower affinity for corticosterone and a shorter half-life than CBG. PMID- 3029260 TI - Differential modulation by Ca2+ of iodide transport processes in a cultured rat thyroid cell strain. AB - The interrelationship between the roles of Ca2+ and cyclic AMP (cAMP) in the control of intracellular iodide accumulation was investigated in the continuous rat thyroid cell strain FRTL-5. Using incubation conditions designed to allow independent study of iodide uptake and efflux into the incubation medium, dibutyryl cAMP (dbcAMP) mimicked the effect of TSH in enhancing iodide uptake, but the responses to both stimulators were decreased in the presence of the Ca2+ ionophore A23187, although the ionophore was ineffective when addition was delayed until after withdrawal of the stimulators. L-Adrenaline, an alpha adrenergic agonist inactive with respect to cAMP accumulation in this cell strain, was also inactive with regard to iodide uptake. The rapid efflux of iodide from FRTL-5 cells seen in response to TSH was not mimicked by dbcAMP, but was reproduced by A23187, providing evidence for the involvement of Ca2+ in the release process. Under the latter conditions, no additional effect of TSH was obtained. L-Adrenaline also stimulated iodide efflux, in support of a possible role of alpha-adrenergic agonists in controlling the apical membrane p presentation of iodide in vivo. The observed differential effects of dbcAMP and alpha-adrenergic agonists on iodide uptake and efflux respectively, provide further evidence for the existence of two functionally distinct mechanisms controlling the available level of intracellular iodide. Opposing effects of free intracellular Ca2+ may, however, serve to control the activities of both influx and efflux processes since, whilst experimental increments in Ca2+ appear to promote iodide efflux through a non-cAMP-mediated pathway.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3029261 TI - Differences in prolactin and LH responses to acute stress between peripuberal and adult male rats. AB - Pituitary-adrenal, pituitary-gonadal and prolactin responses to acute stress (restraint) were studied in peripuberal and adult male rats. The pituitary adrenal response to restraint stress did not differ in peripuberal and adult rats. Prolactin increase during stress was less marked in peripuberal animals. While an increase in LH during stress was observed in adult rats, peripuberal animals did not respond to stress. Testosterone levels were also lower in peripuberal than in adult rats. Diminished LH and prolactin responses to stress in peripuberal rats did not appear to be due either to increased pituitary adrenal activity or to altered pituitary responsiveness to LHRH and dopamine respectively. Peripuberal rats were also more sensitive to the action of morphine on LH and prolactin release than were adult rats, suggesting that endogenous opioids may be involved in the LH and prolactin responses to acute stress. Differences in the maturation of central mechanisms rather than in pituitary response appear to be responsible for the differing responses to acute stress. PMID- 3029263 TI - Adrenal synthesis of corticosterone in response to ACTH in rats is influenced by leukotriene A4 and by lipoxygenase intermediates. AB - The possible involvement of the lipoxygenase pathway of arachidonic acid metabolism in the events which take place during ACTH-induced stimulation of corticosterone secretion has been studied using an isolated rat adrenal cell system. Incubation with arachidonic acid resulted in an inhibition of ACTH stimulated corticosterone production. The lipoxygenase pathway inhibitors nordihydroguaretic acid (NDGA), eicosatetraynoic acid (ETYA) and compound BW755C also produced inhibition of ACTH-stimulated corticosterone synthesis. The concentrations of the inhibitors at which 50% inhibition occurred were 15, 34 and 37 mumol/l respectively. The inhibitions produced by NDGA and ETYA were independent of cyclic AMP output. NDGA also inhibited corticosterone production induced by dibutyryl cyclic AMP but had no effect on corticosterone synthesis induced by pregnenolone. Preincubation of adrenal cells with the lipoxygenase products 5, 12 and 15 hydroxyeicosatetraenoic acid (HETE) and with leukotrienes A4, B4, C4, D4 and E4 resulted in significant inhibitions of corticosterone production in response to ACTH with leukotriene A4 (LTA4) and with 15HETE and 5HETE. Conversely, incubation with glutathione (GSH), which is known to reduce intracellular LTA4 levels, produced stimulation (at 5 mmol GSH/l) and inhibition (at 50 mmol GSH/l) of corticosterone output. These studies suggest that the lipoxygenase pathway may be involved in ACTH-stimulated corticosterone synthesis. PMID- 3029262 TI - Identification, properties and effect of hormones on rat preputial gland peroxidase. AB - A highly active soluble peroxidase has been identified in the preputial glands of the rat. The enzyme was detectable in the sebaceous secretion of the glands and showed catalytic properties characteristic of true peroxidase. It had a native molecular weight of around 73,000 as determined by gel-permeation studies. Immunologically the enzyme cross-reacted with an antiserum against bovine lactoperoxidase. Administration of progesterone resulted in a significant increase in the total activity of the enzyme, while testosterone and oestradiol had no such effect. The enzyme had a similar molecular weight and similar catalytic and immunological properties to rat uterine fluid peroxidase but differed markedly in respect to sensitivity to oestradiol. PMID- 3029264 TI - Extraction and characterization of a cytochemically assayable Na+/K+-ATPase inhibitor/glucose-6-phosphate dehydrogenase stimulator in the hypothalamus and plasma of man and the rat. AB - Some physicochemical properties of partially purified hypothalamic material from the spontaneously hypertensive rat, and of plasma from man and the rat, have been characterized using a validated cytochemical bioassay which measures the ability of biological fluids to stimulate fresh guinea-pig kidney glucose-6-phosphate dehydrogenase (G6PD) after 2 min of exposure to the test substance, as an indication of their ability to inhibit Na+/K+ adenosine triphosphatase (Na+/K+ ATPase) after 4-6 min of exposure. The G6PD-stimulating activity of both hypothalamic extract and plasma is soluble in water and insoluble in chloroform. During electrophoresis the activity from both sites appears in the same fractions and travels considerably further than lysine. After high-pressure liquid chromatography the activity of hypothalamic extract appears in a discreet fraction which does not absorb u.v. light. The activity of both the hypothalamic extract and plasma survives boiling and acid hydrolysis, but is substantially inhibited by prior incubation with digoxin antibody. From ultrafiltration studies, the substance responsible for the ability to stimulate G6PD appears to have a molecular weight of less than 500. The G6PD-stimulating activity of hypothalamic extracts was destroyed by ashing and by base hydrolysis. The ability of plasma of high activity to stimulate G6PD is considerably increased by incubating at 37 degrees C for 15 min and destroyed by incubation for 45 min. It is concluded that these and several other previously noted similarities suggest that the cytochemically assayable Na+/K+-ATPase-inhibiting/G6PD-stimulating activity in the plasma and hypothalamus may be due to the same ouabain-like substance. PMID- 3029265 TI - Ethan-1,2-dimethanesulphonate reduces testicular oxytocin content and seminiferous tubule movements in the rat. AB - An oxytocin-like peptide is present in the interstitial cells of the testis, and testicular concentrations of oxytocin have been shown to increase seminiferous tubule movements in vitro. We have used the drug ethan-1,2-dimethanesulphonate (EDS), which depletes the Leydig cell population of the adult rat testis, to examine further the relationships between the Leydig cell, testicular oxytocin and tubular movements. Adult rats were injected i.p. with a single dose of EDS (75 mg/kg) or of vehicle (25% dimethyl sulphoxide). Histological study 3 and 10 days after treatment with EDS showed a reduction in the number of interstitial cells, and levels of oxytocin immunoreactivity were undetectable by radioimmunoassay. Immunostaining revealed very few oxytocin-reactive cells. Spontaneous contractile activity of the seminiferous tubules in vitro was also dramatically reduced, but could be restored by the addition of oxytocin to the medium. Four weeks after EDS treatment, the interstitial cells were similar to those in the control animals both in number and in immunostaining; immunoassayable oxytocin was present and tubular movements were normal. The EDS effect, seen at 3 and 10 days, was not altered by daily treatment with testosterone. However, repopulation of the testes with oxytocin-immunoreactive cells was not seen until 6 weeks in the testosterone-treated animals. We suggest that the Leydig cells are the main source of oxytocin immunoreactivity in the testis and that this oxytocin is involved in modulating seminiferous tubule movements and the resultant sperm transport. The results also imply that testosterone does not play a major role in controlling tubular activity in the mature rat. PMID- 3029266 TI - Generation of diversity in T cell receptor repertoire specific for pigeon cytochrome c. AB - 17 T cell clones and 3 T cell lines, specific for pigeon cytochrome c, were analyzed for fine specificity and rearranged T cell receptor (TCR) gene elements. Clones of similar fine specificities were grouped into one of four phenotypes, and correlations between phenotype differences and gene usage could be made. All the lines and clones rearranged a member of the V alpha 2B4 gene family to a limited number of J alpha regions. The beta chain was made up of one of three non cross-hybridizing V beta regions, each rearranging to only one or two J beta s. The use of alternate V beta regions could be correlated with phenotype differences, which were manifested either as MHC- or MHC and antigen-specificity changes. In addition, the presence of alloreactivity, which defined a phenotype difference, could be correlated solely with the use of an alternate J alpha region. These observations were substantiated by prospective analyses of pigeon cytochrome c-specific T cell lines that were selected for alternate MHC specificity or alloreactivity and were found to express the correlated alpha and beta chain rearrangements. Previously, the TCR DNA sequences from two clones, each representing a variant of one phenotype, showed sequence differences only in the N regions of their TCR genes. Since only these two variants, using identical V alpha-J alpha and V beta-J beta gene elements, were repeatedly observed in this study, we would predict that the junctional diversity differences are selectable. In this T cell response, all the gene elements involved in the generation of diversity appear to be selected, and may therefore be important in the determination of TCR specificity. This high degree of receptor gene selection represents a fundamental difference from the diversity seen in several extensively analyzed antibody responses. PMID- 3029268 TI - Identification of viral molecules recognized by influenza-specific human cytotoxic T lymphocytes. AB - Human cytotoxic T cells specific for influenza A virus were tested for recognition of each of the ten influenza A virus proteins expressed in target cells using recombinant vaccinia viruses. They recognized the matrix M1, polymerase PB2, and nucleoproteins of influenza virus in association with MHC class I antigens. These internal viral proteins were seen by CTL in conjunction with one or more of the available dependent HLA gene products. There was no detectable recognition of influenza virus surface glycoproteins in target cells. PMID- 3029267 TI - Antigen presentation by chemically modified splenocytes induces antigen-specific T cell unresponsiveness in vitro and in vivo. AB - We investigated the antigen specificity and presentation requirements for inactivation of T lymphocytes in vitro and in vivo. In vitro studies revealed that splenocytes treated with the crosslinker 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (ECDI) and soluble antigen fragments failed to stimulate significant proliferation by normal pigeon cytochrome c-specific T cell clones, suggesting that the chemical treatment inactivated full antigen presentation function. However, T cell clones exposed to ECDI-treated splenocytes and antigen in vitro were rendered unresponsive for at least 8 d to subsequent antigen stimulation with normal presenting cells. As predicted by the in vitro results, specific T cell unresponsiveness was also induced in vivo in B10.A mice injected intravenously with B10.A, but not B10.A(4R), splenocytes coupled with pigeon cytochrome c via ECDI. The antigen and MHC specificity of the induction of this T cell unresponsiveness in vitro and in vivo was identical to that required for T cell activation. These results suggest that nonmitogenic T cell recognition of antigen/MHC on ECDI-modified APCs results in the functional inactivation of T cell clones. PMID- 3029269 TI - Simian virus 40 (SV40)-transgenic mice that develop tumors are specifically tolerant to SV40 T antigen. AB - The ability to mount an immune response to simian virus 40 (SV40) T antigen was evaluated using mice from two distinct SV40 transgenic lines derived from injection of the same gene construct. Our studies demonstrate functional immune tolerance to SV40 T antigen in a SV40 transgenic line that consistently develops tumors of the choroid plexus by 7 mo of age. Antibodies to SV40 T antigen are undetectable in the serum of these animals; furthermore, mice from this line are unable to generate SV40-specific CTL after primary or secondary immunization with the virus, although they mount a normal CTL response to vaccinia virus when appropriately immunized. In contrast, we find that mice from a second transgenic line of low tumor incidence can mount a humoral response to SV40 T antigen, and upon immunization they generally respond with a vigorous cytotoxic T cell response to SV40 T antigen. These data suggest that specific immune tolerance to the product of an integrated viral oncogene may be induced, and is likely a reflection of the time in development at which the gene product first appears. Immune tolerance or responsiveness to the endogenous oncogene product may in turn play a role in the tumorigenic potential of such genes. PMID- 3029270 TI - A synthetic peptide induces long-term protection from lethal infection with herpes simplex virus 2. AB - Immunization against viral pathogens is generally directed toward the induction of virus neutralizing antibody (VNA) and the maintenance of the potential for a second-set (IgG) response. Indeed, an elevated level of specific antibody is considered a reliable clinical indicator that a state of immunity exists in the host. However, in the case of herpes simplex virus (HSV), the presence of circulating VNA does not necessarily correlate with protection. Thus, it has been found that secondary infections occur in individuals even with high neutralizing titers to HSV, suggesting that antibody to the virus may be useless or even deleterious. In consideration of these facts, we were interested in inducing a T cell response to HSV. We had already shown that synthetic peptides corresponding to the NH3-terminal region of the glycoprotein D (gD) molecule of HSV could induce a strong T cell response when injected into mice, but did not, by themselves, confer protection. In this report, we examined the ability of peptides, covalently coupled to palmitic acid and incorporated into liposomes, to induce virus-specific T cell responses that confer protection against a lethal challenge of HSV-2. We have demonstrated that long-term protective immunity is achieved with a single immunization in the absence of neutralizing antibody when antigen is presented in this form. Furthermore, T cells but not serum from such immune mice can adoptively transfer this protection. PMID- 3029271 TI - A murine hybridoma with large cytoplasmic inclusions of kappa light chains. AB - A murine hybridoma cell line has been established that consistently forms large cytoplasmic inclusions. These structures bind antibody specific for mouse kappa L chain when stained in situ. SDS-PAGE analysis of isolated inclusion bodies produce a single protein band of approximately 26,000 Mr that reacts with anti kappa antibody when transferred to nitrocellulose. No carbohydrate was detected in association with the purified protein. These data are consistent with the intracellular retention and deposition of complete kappa L chain protein. PMID- 3029272 TI - In vivo application of recombinant interleukin 2 in the immunotherapy of established cytomegalovirus infection. AB - We have shown in a murine model system for cytomegalovirus (CMV) disease in the immunocompromised host that in vivo application of recombinant human IL-2 (rhIL 2) can enhance the antiviral effect of a limited number of CD8+T lymphocytes, not only in prophylaxis, but also in therapy, when virus has already colonized host tissues. The observed net effect of IL-2 was consistent with the assumption of daily effector population doublings. The prospects for IL-2-supported immunotherapy of established CMV infection depend upon the tissues involved in disease. It appears that the prospects for controlling established CMV adrenalitis are less promising than for a therapy of interstitial CMV pneumonia. PMID- 3029273 TI - Enolase isoenzymes as tumour markers. AB - alpha- and gamma-enolase isoenzyme substance concentrations were measured in serum and plasma from healthy subjects and from 174 patients with different solid tumours. While alpha-enolase was found to be increased in the plasma of patients with tumours of quite different origin, gamma-enolase apparently reflected malignancies of the neuroendocrine system. Before the beginning of the cytotoxic therapy gamma-enolase was increased above the upper limit of the reference range (10 micrograms/l) in 27/27 patients (100%) suffering from small cell lung cancer. Most patients with squamous cell carcinoma of the lung or with prostatic cancer exhibited normal gamma-enolase, while both tumour types produced high plasma substance concentrations of the alpha-isoenzymes of enolase. PMID- 3029274 TI - Computerized 99mTc-pertechnetate thyroid uptake measurement--development of a clinically useful system. PMID- 3029275 TI - Mass screening of hepatobiliary diseases by real-time ultrasonography. PMID- 3029276 TI - Studies on regional hepatic blood flow and substrate metabolism in patients with hepatocellular carcinoma. PMID- 3029277 TI - Effects of extracellular cations on translation in poliovirus-infected cells. AB - The effect of pH and different concentrations of added monovalent and divalent cations on translation in poliovirus-infected HeLa cells has been examined. A strong effect on protein synthesis was observed when the concentration of sodium ions was modified. If cells were placed in a hypotonic medium after virus adsorption, no shut-off of cellular translation took place, nor were viral proteins synthesized. An increase in the multiplicity of infection partially overcame this effect. Reversal of the shut-off of cellular translation and inhibition of viral protein synthesis was achieved when cells were placed in hypotonic medium 2 h after infection. Modification of divalent cations (calcium or magnesium) had little or no effect on the pattern of translation. On the other hand, acidic pH (below 6) inhibited both cellular and viral protein synthesis, whereas basic pH had no influence. During infection the synthesis of poliovirus proteins reached a maximum at about 4 to 6 h and then declined. This inhibition of viral translation was partially prevented if cells were placed in a medium containing a high concentration of potassium although the cytopathic effect was prevented. These results indicate that viral protein synthesis and the cytopathic effect were, to a large extent, influenced by the external monovalent ion concentration. PMID- 3029278 TI - Cation content in poliovirus-infected HeLa cells. AB - Poliovirus-infected HeLa cells increased their permeability to monovalent ions from the third hour post-infection. At that time infected cells lost their content of potassium ions as measured by efflux of the potassium analogue 86Rb+. The rate of release of 86Rb+ from cells increased as infection proceeded, and was not sensitive to ouabain or quinidine, inhibitors of the Na+/K+ ATPase and Ca2+ induced K+ release system, respectively. The leakage of 86Rb+ was only slightly sensitive to furosemide, an inhibitor of the Na+/K+/Cl- contransport system, suggesting that the mechanism of release involved an increased passive permeability of the cell membrane. The calcium content or its efflux did not vary significantly in the infected cells. Neither were there alterations in the intracellular pH throughout infection. PMID- 3029279 TI - Molecular epidemiology of infectious bronchitis virus in The Netherlands. AB - Twelve Dutch isolates and the M41 strain of infectious bronchitis virus (IBV), a coronavirus of chickens, were characterized by cross-neutralization and T1 finger printing to elucidate their evolutionary relationship. The T1 fingerprinting showed that the Dutch isolates formed two clusters. The first cluster contained strains H52, H120, D387, V1259, V1385 and V1397; the estimated sequence homology is 99%. Cluster two comprised strains D207, D274, D212, D1466, D3128 and D3896, which have about 95% sequence homology. The M41 virus did not belong to either cluster. The four different serotypes which arose in the late 1970s belonged to cluster two and appeared to be different from the vaccine strains (H52 and H120) used at that time. This indicates that the strains were newly introduced and could have arisen from a common virus. On the other hand, three recently isolated field strains were genetically closely related to the vaccine strains H120 and H52 (cluster one), suggesting that these live vaccine strains themselves could have given rise to these serologically altered field isolates. The data are relevant to the development of new vaccine strategies. PMID- 3029280 TI - Characterization of novel viral polyproteins detected in cells infected by the flavivirus Kunjin and radiolabelled in the presence of the leucine analogue hydroxyleucine. AB - Vero cells were infected by Kunjin virus and radiolabelled in the presence of the leucine analogue, threo-beta-hydroxy-DL-leucine (THL). This analogue is known to prevent preprotein processing in cell-free systems when incorporated into signal peptides. Novel Kunjin virus-specified proteins were detected, namely gp140, p120 and gp92; the designations for the proteins indicate their approximate Mr X 10( 3) and whether they are glycosylated. The glycoproteins gp140 and gp92 were observed in cells labelled with [3H]mannose and bound to concanavalin A. Limited proteolytic digestion of gp140, gp92 and gp66 (related to the envelope protein E), and tryptic peptide mapping of these three glycoproteins and p120 indicated that all four were closely related. The glycoproteins gp140 and gp92 were also detected in cells infected by West Nile virus radiolabelled in the presence of THL. Other effects of THL in Kunjin virus-infected cells were a reduction in the incorporation of radioactive amino acids or mannose, a decrease in the yield of haemagglutinin and infectious virus, an inhibition of processing of gp66 to gp53 and an apparent inhibition of synthesis of P98 and P71. We suggest possible explanations for the THL-induced changes in infected cells based on recent models proposed for the synthesis of the yellow fever and West Nile viral proteins. PMID- 3029281 TI - Semliki Forest virus-induced, immune-mediated demyelination: adoptive transfer studies and viral persistence in nude mice. AB - Adoptive transfer experiments in athymic nude mice demonstrated that the demyelination seen in the central nervous system (CNS) following Semliki Forest virus (SFV) infection was directly dependent upon sensitized T lymphocytes. Antibodies generated during the infection did not seem to be involved in the demyelination, but thymus-dependent antibodies (IgG) were responsible for the reduction of brain virus titres. In the absence of a T cell response and T cell dependent antibody production, virus persisted in the CNS for several months. Despite persistence of high virus titres for this time, only mice eventually developing a CNS inflammatory response developed lesions of demyelination. In the absence of an inflammatory response no demyelination was apparent even after several months of persistent infection. Administration of anti-SFV hyperimmune serum intracerebrally to both infected and control mice did not produce demyelination but resulted in CNS tissue degeneration with marked pycnosis. PMID- 3029282 TI - Entry pathway of vesicular stomatitis virus into different host cells. AB - A biochemical and morphological investigation of the mechanism of entry of vesicular stomatitis virus (VSV) into host cells of mammalian (HeLa), avian (CER), piscine (EPC) and arthropod (Aedes albopictus) origin, is described. VSV was capable of infecting all cell lines tested by a endosome- and/or a lysosome dependent step since ammonium chloride and amantadine blocked the early stages of infection. Complement-dependent immune lysis of infected host cells provided evidence that in none of the four different cell types examined did insertion of VSV antigens occur from the outside to any great extent on the cell surface. When the entry process was studied by electron microscopy, virus particles were seen to be bound to the cell surface at 0 degrees C. After warming at 37 degrees C for homeothermic cells or at 26 degrees C for poikilothermic cells, virus was detected within coated pits and coated vesicles and, later, in lysosomes. VSV entry was seen to take place by endocytosis in all four cell lines, which were derived from phylogenetically unrelated species. PMID- 3029283 TI - Glycoproteins of human parainfluenza virus type 3: affinity purification, antigenic characterization and reconstitution into lipid vesicles. AB - Monoclonal antibodies to the envelope glycoproteins, HN and F, of human parainfluenza virus type 3 were coupled to a Sepharose 4B matrix and used for affinity purification of the viral glycoproteins. The purity of the glycoproteins was demonstrated by SDS-PAGE followed by fluorography or silver staining. The antigenicity of the glycoproteins was determined by immunization of rabbits; polyclonal rabbit antisera demonstrated inhibition of functional activities of the virus glycoproteins. The F glycoprotein, when reconstituted into lipid vesicles, showed distinct spike-like projections similar to those of intact virions. PMID- 3029284 TI - Human papillomavirus type 16 DNA from a vulvar carcinoma in situ is present as head-to-tail dimeric episomes with a deletion in the non-coding region. AB - A number of genital cancer biopsy samples were screened for the presence of human papillomavirus type 16 (HPV-16) DNA sequences. One of these samples (a vulvar carcinoma in situ) was found to contain more than 100 copies of HPV-16 DNA sequences per cell. Using this tumour DNA, a genomic library was constructed in bacteriophage lambda and the library was screened for recombinant phage containing HPV-16 sequences. Five recombinant phage clones were isolated and their DNA was analysed by restriction endonuclease digestion and blot hybridization. All five recombinants contained two copies of the HPV-16 genome present in a head-to-tail arrangement. The data are consistent with the presence of HPV-16 sequences in the tumour DNA arranged as genomic dimers in a circular episomal configuration. The HPV-16 genomes contained a deletion within the non coding region, a region which includes the viral origin of DNA replication and transcriptional control sequences. Possible consequences of this deletion for viral replication and transcription are discussed. PMID- 3029285 TI - Virus-lymphocyte interactions during the course of immunosuppressive virus infection. AB - Malignant rabbit fibroma virus (MV) is a lymphocytotropic leporipoxvirus which produces profound immunological dysfunction and lethal fibromyxosarcoma. We examined virus recovery from splenic lymphocytes as a function of time after inoculation in vivo, and correlated this with both immunological function and expression of virus-induced host suppressor activity. MV was most abundant in lymphocytes obtained 4 days following inoculation. At that time, immune function was relatively normal and host suppressor activity was not observed. By 7 days after infection, when active host immunosuppressor functions were observed, virus recovery was decreased. Eleven days post-inoculation host immune function began to recover despite increasing virus-induced tumours and developing opportunistic infection. Simultaneously, MV was no longer recoverable from spleen cells. Spleen cells from day 11 tumour-bearing rabbits did not support MV replication as efficiently as did normal or day 4 or 7 splenic lymphocytes, but they did not alter the ability of MV to grow in the latter cells. By fluorescence examination and cytofluorography, splenic lymphocytes bearing MV antigens were abundant 7 days after infection but disappeared by 11 days. This was temporally related to production of neutralizing antibody to MV, and development of virus-specific lymphocyte proliferative activity. The composition of splenic lymphocytes changed as well: the normal ratio of about 1:1 for B and T cells changed to 1:2 by day 7, and then inverted to almost 2:1 by day 11. Rabbits infected with MV thus appear to recover their immune function, concurrently eliminate virus-infected lymphocytes, and elaborate high titres of neutralizing serum antibodies despite progressive infections and tumour development. PMID- 3029286 TI - Proteins antigenically related to peptides encoded by the mouse mammary tumour virus long terminal repeat sequence are associated with intracytoplasmic A particles. AB - Intracytoplasmic A particles (CAP), previously identified as cytoplasmic nucleocapsid precursors to mouse mammary tumour virus (MMTV), reacted strongly in immunodiffusion tests with polyclonal antibodies raised against synthetic oligopeptides derived from the open reading frame (ORF) in the long terminal repeat (LTR) of MMTV. In Western blots, several CAP proteins (p80, p72-68, p36, p32, p18-12) were reactive with polyclonal antibodies raised against three separate LTR ORF synthetic peptides. Disrupted MMTV virions did not react with the anti-LTR ORF peptides suggesting that ORF proteins were excluded from mature virions during maturation. Serial dilution of anti-LTR ORF antibody demonstrated that the most reactive CAP proteins in Western blots migrated as a doublet band with estimated molecular weights of 68,000 and 72,000. Reactivity of anti-LTR ORF serum with these and other CAP proteins was removed upon preincubation with free synthetic peptide. Absorption with LTR synthetic peptides did not affect the reactivity of antibodies directed against MMTV gag proteins with similarly sized CAP polyproteins. LTR ORF-related proteins with molecular weights similar to those associated with CAP were also detectable in Western blots of total cytoplasmic extracts of MMTV-infected mammary tumour cells. PMID- 3029287 TI - Expression of a provirus of human T cell leukaemia virus type I by DNA transfection. AB - We isolated the full length provirus of human T cell leukaemia virus type I (HTLV I) from MT-2, a lymphoid cell line producing HTLV-I. In three non-lymphoid cell lines (COS7, human osteosarcoma HOS cells, and HeLa) this provirus expressed a trans-acting activity after co-transfection with a recombinant plasmid carrying a bacterial chloramphenicol acetyltransferase gene under the control of a long terminal repeat of HTLV-I provirus. The trans-acting protein p40 was detected by immunoprecipitation in transfected HOS cells. Structural proteins of HTLV-I, the gag and env products, were also formed and processed in the same manner as observed in MT-2 cells. In transfected HeLa cells, the p40 protein was mainly localized in the nucleus, while other structural proteins were detected in the cytoplasm and/or the membrane by indirect immunofluorescence. Syncytium formation was observed in HeLa cells after transfection. These results demonstrated that non-lymphoid cells could produce the major proteins of HTLV-I after DNA transfection of the cloned provirus. PMID- 3029288 TI - Suppression of in vitro neutrophil function by feline leukaemia virus (FeLV) and purified FeLV-p15E. AB - Feline neutrophils (PMN) were isolated and exposed to ultraviolet light inactivated feline leukaemia virus (UV-FeLV) and purified envelope component p15E (FeLV-p15E). Functional capacity of exposed PMN was measured in vitro utilizing the chemiluminescence (CL) response. PMN exposed to UV-FeLV demonstrated depressed CL responses to Ca2+-ionophore A23187 and latex particles. However, FeLV-p15E produced significant suppression in the CL response to A23187 but failed to produce significant alterations in response to latex particles. The data indicate that FeLV-p15E may, in part, be responsible for increased morbidity and mortality among FeLV-infected cats through suppression of the PMN population. PMID- 3029289 TI - Construction and characterization of deletion mutants of pseudorabies virus: a new generation of 'live' vaccines. AB - Various deletions were introduced into a cloned subgenomic fragment (BamHI-7), located in the unique short (US) region of the DNA from the virulent Northern Ireland Aujeszky-3 (NIA-3) strain of pseudorabies virus (PRV). In the cloned HindIII-B fragment, the MluI-BglII fragment was replaced by different MluI-BglII fragments of the deleted BamHI-7 clones. Transfection of the deleted HindIII-B fragments together with the HindIII-A fragment of either the NIA-3 or the non virulent NIA-4 strain yielded replication-competent deletion mutants. The region in US in which sequences were deleted specified several mRNAs. Some of the mRNAs present in cells infected with NIA-3 were absent from cells infected with the deletion mutants, whereas other differently sized mRNAs were generated. The mutants were examined with respect to their biological properties in cell culture, mice and pigs. The results showed that the type of cytopathic effect induced in cell culture seemed to be determined by the UL region, using the mean time to death in mice as a parameter, markers for virulence were present in the US and UL regions and the introduction of deletions in US strongly reduced the virulence of PRV for pigs. Despite the impaired capacity of the deletion mutants to induce high titres of neutralizing antibodies in the serum, inoculation with mutants derived from NIA-3 prevented clinical disease in pigs upon challenge with the virulent parent strain. These deletion mutants provide a good basis for the production of bioengineered live PRV vaccines. PMID- 3029290 TI - Restricted replication of herpes simplex virus type 1 in murine embryonal carcinoma cells. AB - Herpes simplex virus type 1 (HSV-1) has a broad host range but the KOS strain of HSV-1 did not replicate efficiently in murine embryonal carcinoma (EC) cells. The yield of infectious HSV-1 from EC cells was 100- to 1000-fold lower than that from fibroblast cell lines of mouse, monkey or human origin. The thymidine kinase (TK) gene of HSV-1 is expressed early during the infectious cycle. The levels of TK mRNA and of TK activity in infected EC cells were only two- to threefold lower than levels from infected fibroblast cells. Infected EC cells supported replication of about half as much HSV-1 DNA as did fibroblast cells. The reduced yield of infectious virus was consistent with a paucity of virions in infected EC cells examined by electron microscopy, suggesting a major block late during the HSV-1 infectious cycle. We isolated a variant strain of HSV-1, called KOSEC, which replicated as efficiently in EC cells as in mouse fibroblasts. KOSEC infected EC and fibroblast cells, synthesized more TK mRNA, more TK enzyme, and more HSV-1 DNA than did the same cells infected with the KOS stain. Both HSV-1 strains induced similar levels of synthesis of gD, an early viral glycoprotein. By co-infection of EC cells with the KOS and KOSEC virus, both the elevated virus yield and the elevated TK synthesis seen in KOSEC-infected cells appeared to be recessive. Apparently a viral mutation that affects expression of some early viral functions can also overcome the EC cell restriction to HSV-1 replication. PMID- 3029291 TI - Macaque monkey type D retrovirus replicates in vitro in a distinct subpopulation of B lymphocytes. AB - Type D retroviruses have recently been shown to induce a wasting syndrome with associated lymphadenopathy, thymic atrophy and transient decreased peripheral blood lymphocyte blastogenic responsiveness in juvenile macaque monkeys. The replication in vitro of D/New England virus was assessed in various lymphocyte subpopulations to determine the possible pathogenesis of the immune dysfunction induced by this virus. While D/New England did not replicate in cultured T lymphocytes or induce any demonstrable dysfunction of T cells in vitro, it did grow in the cells of the B lymphocyte lineage. D/New England growth occurred in vitro in African Burkitt's lymphoma and pre-B cell lines, but not in Epstein-Barr virus-transformed normal B lymphocytes. The infection of a restricted B lymphocyte population by this primate type D retrovirus may play a role in the aetiology of the immune abnormalities which it induces. PMID- 3029292 TI - Ultrastructural studies of Kunjin virus-infected Aedes albopictus cells. AB - Ultrastructural changes in Aedes albopictus cells infected with Kunjin virus were characterized from 12 to 72 h post-infection. Early in infection (16h), there were no prominent ultrastructural changes except for an increase in the number of vacuoles in the cytoplasm. As the infection progressed the rough endoplasmic reticulum appeared to lengthen and whorls of fibres were observed within some vacuoles. Virus particles were observed in small numbers scattered in the cytoplasm between 24 to 30 h after infection. However, by 48 h, large numbers of morphologically mature virus particles were visible, usually in association with the rough endoplasmic reticulum, with the clusters of vesicles or with convoluted smooth membranes. Although no definite evidence of precursor particles or of budding of virions was obtained, the clusters of vesicles and the various membranous structures appeared to play a role in the morphogenesis of this virus. The results are compatible with the hypothesis that maturation of virus particles occurs within all these virus-induced structures. PMID- 3029293 TI - Integration and transcription of human papillomavirus type 16 and 18 sequences in cell lines derived from cervical carcinomas. AB - Five cell lines, SKG-I, SKG-II, SKG-IIIb, QG-U and QG-H derived from cervical carcinomas of Japanese patients, were examined for the presence of human papillomavirus (HPV) DNA and the expression of viral mRNA. The DNA of HPV type 16 was shown to be linked covalently with SKG-IIb, QG-U and QG-H cell DNA, and HPV 18 DNA with SKG-I and SKG-II cell DNA. Although different regions of the HPV genome were integrated in these cell lines, the non-coding region and an early region including the E6 and E7 open reading frames (ORFs) were conserved in all cell lines. The complete genome of HPV 16 was found in QG-H cells by digestion of the DNA with a single-cut restriction enzyme. The other early region ORFs E1, E2, E4 and E5 were interrupted by flanking host cell DNA, suggesting that the integration into host cell DNA occurs preferentially in this region. HPV-specific mRNA species were detected in all five cell lines. In the three cell lines containing the HPV 16 genome, mRNAs hybridized with the early region of the genome, covering the entire E6 and E7 ORFs and a minor part of the E1 ORF, although the amount and size of the major mRNAs varied in these cell lines. These mRNAs did not hybridize with the late region of the HPV genome containing the L1 and L2 ORFs. In SKG-II, SKG-IIIb and QG-H cells we also detected c-myc and c-Ha ras mRNA expression at about nine times the level of that in normal cells. PMID- 3029294 TI - Synthesis in Escherichia coli and immunological characterization of a polypeptide containing the cleavage sites associated with trypsin enhancement of rotavirus SA11 infectivity. AB - About 45% of the rotavirus SA11 VP3 gene was inserted into a thermoinducible expression plasmid under the control of phage lambda PL promoter. The primary translation product predicted on the basis of the plasmid construction was a hybrid protein in which the 98 amino-terminal amino acids of phage MS2 polymerase were followed by amino acids 42 to 387 of the VP3 protein, which included the region containing the cleavage sites associated with trypsin enhancement of infectivity. On induction, a polypeptide that had the expected mol. wt. and contained VP3-related amino acid sequences as judged by immunological criteria, was synthesized to a level representing about 15% of the total bacterial protein. When a bacterial lysate enriched for the fusion polypeptide was injected into mice, it induced antibodies which inhibited haemagglutination and neutralized SA11 rotavirus infectivity. PMID- 3029295 TI - Serum antibody responses to individual viral polypeptides in human rotavirus infections. AB - A radioimmunoprecipitation assay (RIPA) was used to study the serum antibody responses to individual polypeptides that developed after infection with viruses from human rotavirus subgroups I and II. Paired sera from eight children (1 to 8.5 years of age) were used in the study. Although all of the eight acute sera were negative by the complement fixation test, four of them were positive by RIPA, indicating a previous infection by rotavirus. A significant difference in the number of polypeptides immunoprecipitated was seen among the convalescent sera. The number of polypeptides immunoprecipitated was found to be related to previous infection experience. At most, ten different polypeptides were immunoprecipitated: seven structural polypeptides VP1 to VP7 and three non structural polypeptides, NS1, NS2 and NS3. No sera immunoprecipitated VP8 or VP9. Acute sera positive by RIPA immunoprecipitated up to five polypeptides, VP1, VP2, VP3, VP4 and VP6. One of the non-structural proteins (NS2) was found to be particularly immunogenic, since antibodies to this polypeptide were detected in several convalescent sera. Among the structural proteins VP2 and VP6 were found to be the two immunodominant polypeptides which were recognized by all convalescent sera. Only three convalescent sera immunoprecipitated VP7, the major type-specific antigen responsible for inducing neutralizing antibodies. Three of four originally seronegative children with no reactivity in the convalescent sera to VP7 developed neutralizing antibodies to a single serotype. One child developed antibodies to two serotypes. PMID- 3029296 TI - Isolation and characterization of two group A rotaviruses with unusual genome profiles. AB - Analysis of the genomic dsRNA of rotaviruses isolated from calves with subclinical infections has revealed eight calves excreting group A viruses with unusual genome profiles. Two of these virus strains, C7-176 and C7-183, were grown and cloned by plaque purification in cell culture. Examination of their genome profiles after cloning showed that the unusual pattern had been retained. They were without RNA segment 11 but had an extra band (6a) migrating between segments 6 and 7. However, they contained the triplet of segments 7, 8 and 9, of similar size, which is characteristic of group A rotaviruses. The number and mol. wt. of the intracellular polypeptides induced by these viruses were similar to those of the bovine group A rotavirus UK strain. Analysis of the RNA transcripts produced by the transcription in vitro of purified C7-183 virus showed that segment 6a produced a large RNA transcript of corresponding size. After isolation, this transcript was translated in a rabbit reticulocyte lysate preparation and yielded a single polypeptide, vp11, equivalent to the product of segment 11 of rotaviruses with typical genome profiles. PMID- 3029297 TI - Characterization and physical mapping of Molluscum contagiosum virus DNA and location of a sequence capable of encoding a conserved domain of epidermal growth factor. AB - DNA from Molluscum contagiosum virus (MCV) isolates was analysed by restriction endonuclease cleavage, revealing two virus subtypes. Physical maps of cleavage sites for BamHI, ClaI and HindIII were constructed, and found to differ extensively between the two subtypes. MCV DNA was similar to Orthopoxvirus DNA with respect to size, terminal cross-linking and the presence of inverted terminal repetitions, but did not hybridize with vaccinia virus DNA. The genomes of the two MCV subtypes cross-hybridized and were colinear except for two small regions. There was sequence homology between DNA from corresponding map regions of the MCV subtypes but, in contrast to Orthopoxvirus DNA, no conservation of restriction sites. A synthetic oligonucleotide probe representing a conserved domain of epidermal growth factor, alpha-transforming growth factor and the vaccinia growth factor identified equivalent regions of both MCV genomes as having the potential to encode this domain. This locus is similar to the position of the vaccinia growth factor gene in vaccinia virus DNA. Thus MCV may induce epidermal cell proliferation and tumourigenesis by expression of an epidermal growth factor-like polypeptide. PMID- 3029298 TI - Characterization of a molecular clone of RFM/Un mouse chromosomal DNA that contains a full-length endogenous murine leukaemia virus-related proviral genome. AB - A 12.4 kbp HindIII chromosomal DNA fragment harbouring an apparently intact 9.2 kbp endogenous murine leukaemia virus (MuLV)-related proviral genome was isolated from an RFM/Un strain mouse by molecular cloning and designated pRFM #6. Nucleotide sequence analysis revealed the following characteristic features in the pRFM #6 provirus: a distinct 200 bp sequence in the long terminal repeat (LTR) mid-U3 region, a primer binding site for glutamine tRNA, a 3' pol region encoding an 'endonuclease' protein of 390 amino acids, and the mink cell focus forming virus type-specific sequence at the 5' portion of the env gene. The 699 bp 5' LTR and 700 bp 3' LTR of pRFM #6 provirus were identical except for three base changes in the U3 'enhancer' region. At the cell-provirus DNA junction, 4 bp direct repeats were present. The proviral genome was found at the same chromosomal DNA site in BALB/c, AKR, C3H, CBA and RFM strain mice, but not in NFS/N or C57BL/6 strain mice. PMID- 3029299 TI - Induction of demyelination by a temperature-sensitive mutant of the coronavirus MHV-A59 is associated with restriction of viral replication in the brain. AB - The neurovirulence of eight temperature-sensitive (ts) mutants of mouse hepatitis virus strain A59 in 4-week-old BALB/c mice was investigated. Whereas a dose of 100 p.f.u. of wild-type virus killed mice within a week, a 1000-fold higher dose of ts mutants did not. Three ts mutants induced demyelinating disease in the central nervous system (CNS). The pathology of the demyelinating disease caused by one mutant, designated ts-342, was studied in detail. Pathological changes, starting 3 days post-inoculation (p.i.), were characterized by inflammation and demyelination in the CNS. Antibody responses directed against all virus-specific structural proteins were present at 7 days p.i. No virus particles were observed by electron microscopy at 14 days p.i. However, macrophages and lymphocytes were abundant in the areas of demyelination. The growth kinetics in vivo of wild-type virus, ts-342 and a revertant of ts-342 were compared. Wild-type virus and the revertant replicated rapidly in the brain and spread to the liver causing a lethal hepatitis. Ts-342, however, replicated to a much lesser extent within the brain and could not be detected in the blood or liver. The ts lesion in the genome of ts-342 seems, therefore, to determine the outcome of the infection. PMID- 3029300 TI - Herpes simplex virus glycoproteins associated with different morphological entities projecting from the virion envelope. AB - Spikes of different kinds, distinct in size and appearance were detected on the surfaces of herpes simplex virions by electron microscopy of negatively stained preparations. Use of monoclonal antibodies coupled to colloidal gold permitted identification of viral glycoproteins present in different structures projecting from the virion envelope. Antibodies specific for the glycoprotein designated gB bound to the most prominent spikes, which were about 14 nm long and, in side view, had a flattened T-shaped top. Antibodies specific for gC bound to structures that, in some instances, appeared to extend as much as 24 nm from the surface of the envelope and were too thin to resolve. Antibodies specific for gD bound to structures that extended as much as 8 to 10 nm from the surface of the envelope. The gB spikes were invariably clustered, usually in protrusions of the envelope varying from small bulbous distentions to long tail-like projections. The gC components were randomly distributed and widely spaced and the gD components were irregularly clustered in patterns distinct from those of the gB spikes. These three glycoproteins therefore form structures that are different in size, morphology and distribution in the envelope. PMID- 3029301 TI - DNA-binding proteins of herpes simplex virus type 1-infected BHK cell nuclear matrices. AB - The nuclear matrix is involved in the replicative cycle of herpes simplex virus type 1 (HSV-1) and in at least some cases viral DNA has been shown to be closely associated with this structure. In this communication, we report the presence of five DNA-binding proteins in the nuclear matrix of HSV-1-infected BHK cells. These proteins (p114, p89, p77, p37 and p29) were detected by probing with 32P labelled HSV DNA after Western blotting of nuclear matrix proteins. Three were identified as virion components: p89 as VP12, p77 as VP13 and p37 as the capsid protein VP22a. These polypeptides were detected in cells and nuclei and found to be associated with the nuclear matrix late during the lytic cycle, long after the onset of viral DNA replication. The nuclear matrix-binding capacity of VP22a depended on viral DNA replication, since after DNA polymerase inhibition it was still synthesized and transported into the nucleus but was no longer associated with the nuclear matrix. After inhibition of viral DNA synthesis, VP13 was no longer found in cells, nuclei or nuclear matrices. These results suggest a possible involvement in anchoring viral progeny DNA to the nuclear matrix. PMID- 3029302 TI - Genomic comparison of herpes simplex virus type 1 isolates from Japan, Sweden and Kenya. AB - One-hundred and seventy-two epidemiologically unrelated herpes simplex virus type 1 (HSV-1) strains isolated in Japan (104 strains) and Sweden (68 strains) were compared by analysis of their genome structures using four restriction endonucleases, BamHI, KpnI, SalI and HindIII. In addition, 32 Kenyan HSV-1 isolates previously compared to Japanese isolates were included for further comparison with the Swedish isolates. Remarkable and statistically significant differences were found between the HSV-1 isolates from the three countries. One hundred and thirty cleavage sites were examined, and it was shown that isolates from two of the three countries were statistically distinguishable at 27 of these loci. Pairwise comparison between isolates from Japan and Sweden, Kenya and Sweden, and Japan and Kenya revealed variation in 18, 16 and 23 sites, respectively. By considering gains and losses of 19 sites, the total of 204 strains could be classified into 92 distinct cleavage patterns. Isolates from the three countries could be distinguished from each other by the pattern, except for one Swedish and two Kenyan isolates which shared a pattern. Twenty-one fragments that were present or absent only in individual isolates from one or other of the three countries could be detected. These results show that HSV-1 strains from geographically separate countries or anthropologically different races have distinct distributions of endonuclease recognition sites. PMID- 3029303 TI - A new human cytomegalovirus isolate has an invertible subsegment within its L component producing eight genome isomers. AB - A HindIII cleavage map of the genome DNA of a new isolate of human cytomegalovirus (HCMV), strain Tanaka, was constructed by cosmid cloning and Southern blot hybridization of virion DNA. The genome was found to be unique in that its long (L) component was composed of two subsegments, L1 and L2, and subsegment L2 underwent inversion relative to L1 at high frequency. In addition to the normal inversions of the L and short (S) components, this produced eight genome isomers. The novel invertible subsegment was flanked by an inverted sequence distinct from the inversion-specific a sequence present in the terminal and junction regions of the genome. PMID- 3029304 TI - Cytomegalovirus strain AD169 binds beta 2 microglobulin in vitro after release from cells. AB - We previously reported that a host protein, beta 2 microglobulin (beta 2m) inhibited the detection of human cytomegalovirus (CMV) in urine specimens by enzyme immunoassay and postulated that beta 2m bound to the virus particle and masked the viral antigenic determinants. We report here that CMV strain AD169 grown in cell culture bound human beta 2m when this protein was added to cell culture fluids or when the virus was added to urine. Such binding was not seen with herpes simplex virus. CMV could also bind bovine beta 2m from foetal calf serum in cell culture fluids. The use of radiolabelled beta 2m in other experiments showed that CMV bound beta 2m after release from cells and that the bound beta 2m did not represent acquisition of class I HLA molecules during budding from host cell membranes. Immunoprecipitation studies showed that beta 2m was bound by two viral envelope proteins beta 2m BP1 (beta 2m-binding protein 1) and beta 2m BP2 of molecular masses 36,000 and 65,000 daltons respectively. beta 2m could not bind to separated viral proteins under reducing or non-reducing conditions. We propose that interaction of these two proteins on the viral surface is required to enable CMV to bind beta 2m. PMID- 3029305 TI - Cytomegalovirus in urine specimens has host beta 2 microglobulin bound to the viral envelope: a mechanism of evading the host immune response? AB - We have previously reported that human cytomegalovirus (CMV) from urine specimens cannot be captured onto a solid phase by CMV-specific monoclonal antibodies that can capture CMV grown in vitro. We report here that CMV exists in vivo in body fluids such as urine as beta 2 microglobulin (beta 2 m)-coated particles. We have demonstrated the presence of beta 2m on CMV purified directly from urine by Western blotting and have shown that the beta 2m was associated with the viral envelope. Urinary CMV could be specifically bound by an affinity column comprising a monoclonal antibody specific for beta 2m bound to Sepharose. The beta 2m-coated urinary CMV could not be neutralized by hyperimmune globulin, human immune sera or murine monoclonal antibodies that could neutralize CMV grown in cell culture. We conclude that the binding of beta 2m by CMV masks the important antigenic sites necessary for neutralization which are recognized by man's immune response. We propose that CMV has evolved this mechanism of coating itself in a host protein as a mechanism of evading the host immune response and facilitating transmission between individuals. PMID- 3029306 TI - Beta 2 microglobulin enhances the infectivity of cytomegalovirus and when bound to the virus enables class I HLA molecules to be used as a virus receptor. AB - We have previously demonstrated that human cytomegalovirus (CMV) binds the host protein beta 2 microglobulin (beta 2m) from body fluids or from cell culture media. In this report we have examined the effect of the beta 2m on viral infectivity. We have shown that the addition of human purified beta 2m, or a fraction of foetal calf serum corresponding to bovine beta 2m, to culture medium increased the amount of infectious extracellular CMV, compared to that from cells grown in serum-free medium. Metabolic labelling experiments demonstrated that this effect was not due to an increase in the amount of extracellular virus but to an increase in the infectivity of the virus present in extracellular fluids. We concluded that the binding of beta 2m by CMV increased its infectivity. We have shown that CMV and beta 2m compete for binding sites on fibroblasts. As the main binding site on cells for beta 2m is the class I HLA heavy chain we compared the binding of CMV to the Raji and Daudi cell lines which express or lack expression of class I HLA molecules. The binding of radiolabelled beta 2m-coated CMV was significantly higher to Raji cells than to Daudi cells. Furthermore, CMV could compete with beta 2m for binding to Raji cells, although the reverse was not true. These results demonstrate that CMV can use class I HLA molecules as a virus receptor. We propose that when coated with beta 2m, CMV has the capacity to displace beta 2m from the class I HLA heavy chain-beta 2m dimer on the cell surface and bind to cells. The fact that beta 2m enhances infectivity suggests that such binding leads to productive infection of cells. PMID- 3029308 TI - Herpes simplex virus type 1 latency in rabbit corneal cells in vitro: reactivation and recombination following intratypic superinfection of long term cultures. AB - Herpes simplex virus type 1 (HSV-1) has been isolated from explanted human corneas after cultivation in vitro. To determine whether HSV-1 is persistent or latent in corneal cells, a system to study HSV-1 infection of rabbit corneal cells in vitro was developed. By elevation of the incubation temperature to 42 degrees C before and during HSV-1 infection it was shown that both keratocytes and epithelial cells support a nonproductive rather than a productive infection. On subsequent temperature reduction to 37 degrees C, infectious virus was released from both cell types. Addition of the viral inhibitor acycloguanosine during the last 5 days of a 14 day incubation at 42 degrees C did not reduce the frequency of viral shedding following transfer to 37 degrees C, indicating that corneal cells support a latent as opposed to persistent infection. Cultures which failed to shed virus spontaneously up to 29 days post-inoculation were superinfected at 37 degrees C, with an XbaI site deletion mutant of HSV-1. Restriction endonuclease analysis of progeny identified both the initial infecting virus and recombinants between the parental and superinfecting genomes. PMID- 3029307 TI - Human epithelial cell expression of an Epstein-Barr virus receptor. AB - Cultured human epithelium bound and internalized radiolabelled Epstein-Barr virus (EBV) within 1 h of exposure. A similar percentage of cultured cells also were reactive with monoclonal antibodies to the EBV/C3d receptor of B lymphocytes. In cross-sections of fresh frozen, stratified epithelium, receptor expression seemed limited to the less differentiated subpopulation of cells. These findings support the notion of direct infection of epithelial cells by EBV and suggest a viral life cycle in epithelium dependent on the stage of cell differentiation. PMID- 3029309 TI - Evidence for antigenic stability of tick-borne encephalitis virus by the analysis of natural isolates. AB - Strains of tick-borne encephalitis (TBE) virus isolated from ticks in natural foci in Austria were compared to strains isolated from the same foci 14 years previously. Comparative peptide mapping of the envelope (E) glycoproteins as well as analysis of the antigenic structure of the E proteins by the use of 14 monoclonal antibodies defining different epitopes did not provide evidence for antigenic variation. The same also holds true for isolates from a probably newly established natural focus in Western Austria. These results confirm previous data by showing that under natural ecological conditions TBE virus is quite stable and does not undergo major antigenic changes. PMID- 3029310 TI - Selection of coxsackievirus B4 variants with monoclonal antibodies results in attenuation. AB - Inoculation of suckling mice with coxsackievirus B4 (CB4) results in the death of a majority of the animals. In this study we selected antigenic variants of CB4 in the presence of neutralizing monoclonal antibodies and tested them to see whether they were attenuated. Antigenic variants selected with a single antibody showed little or no attenuation by producing a high mortality (60 to 100%). A double variant selected with two antibodies showed considerable attenuation by causing only 25% mortality. A triple variant selected with three antibodies was almost completely attenuated (killed only 5% of the animals). Polypeptides from these variants were tested for their ability to interact with the monoclonal antibodies used for their selection. These studies showed that resistance of variant virus to neutralization in general was due to the inability of the antibody to bind to the virus. However, one of the antibodies could bind but not neutralize the virus, perhaps due to an alteration in the epitope. It is concluded that selection of CB4 variants using more than one neutralizing monoclonal antibody can lead to attenuation of the virus. PMID- 3029311 TI - Inhibition of BK virus haemagglutination by gangliosides. AB - The effect of gangliosides extracted from human group O Rh+ erythrocytes on haemagglutination by BK virus was investigated. Experiments were performed on both ganglioside mixtures and isolated fractions separated by column chromatography and characterized by thin-layer chromatography. These results were compared with those obtained with standard preparations of gangliosides, and the inhibiting activity was shown to be confined mainly to gangliosides with a RF lower than GM1. It was also observed that the insertion of gangliosides in liposomes increased the haemagglutination-inhibiting activity and that ganglioside coating restored the ability of glycosidase-treated human red blood cells to agglutinate. PMID- 3029312 TI - Comparison of porcine parvovirus to other parvoviruses by restriction site mapping and hybridization analysis of Southern Blots. AB - The genomic relationship between porcine parvovirus (PPV) and several other autonomous parvoviruses was examined by restriction site and hybridization analysis. Restriction site maps of the PPV genome were prepared by digesting the double-stranded replicative form of the viral DNA with each of eight restriction enzymes. Subsequent comparison of such maps with those previously reported for PPV, canine parvovirus (CPV), feline panleukopenia virus (FPV), minute virus of mice (MVM), H-1 virus (H-1) and bovine parvovirus (BPV) revealed that while the maps of CPV, FPV, MVM and H-1 had a number of features in common, those of PPV and BPV were substantially different. For hybridization analysis radioactive probes prepared by nick translation of PPV, CPV and BPV genomes were tested under conditions of both low and high stringency for homologous hybridization and for heterologous hybridization with each of the other two viruses and with FPV. The results of these tests indicated homology among the genomes of PPV, CPV and FPV, but little or no homology between the genome of BPV and those of any of the other viruses tested. Additional tests with restriction fragments of PPV and a CPV probe indicated that heterologous hybridization was confined primarily to a segment of the genome between 1.85 and 2.7 kb from the 3' end. Based on transcriptional maps previously determined for several of the rodent parvoviruses, this interval is likely to include part of the coding sequences for both non-structural and structural proteins and may be the genetic basis for the replicative as well as the antigenic similarities between PPV and both CPV and FPV. PMID- 3029313 TI - Restriction endonuclease analysis of herpes simplex virus from recrudescent lesions, from latent infection and during passage in the skin and nervous system of mice. AB - The stability of the restriction endonuclease profile of herpes simplex virus type 1 strain SC16 in mice was studied. Virus isolated from skin during acute infection was compared with that from latently infected ganglia and with that from recrudescent lesions induced by trauma. In another experiment virus serially passaged only in skin was compared with virus that had also replicated in the nervous system. The loss or gain of specific restriction sites was not observed but in some cases the mobility of certain fragments decreased. PMID- 3029314 TI - Effects of the epipodophyllotoxin VP-16-213 on herpes simplex virus type 2 replication. AB - It has been recently shown that VP-16-213, a semi-synthetic derivative of podophyllotoxin, inhibits the function of mammalian DNA topoisomerase II. In the present study, we examined the effects of VP-16-213 on the replication of herpes simplex virus type 2 (HSV-2). The compound did not inhibit the synthesis of early viral polypeptides at concentrations at which viral DNA synthesis was strongly suppressed, but induced double-strand breaks in newly synthesized HSV DNA. Electron microscopic examination of treated cells revealed the presence of a number of capsids with empty or partial cores. The level of topoisomerase II activity remained unaltered after infection, and all attempts to isolate VP-16 213-resistant mutants of HSV-2 have failed in spite of extensive efforts. It is suggested therefore that the mode of action of VP-16-213 may be inhibition of viral DNA synthesis by impairing the function of host cell topoisomerase II. PMID- 3029315 TI - A recombinant human interferon-alpha B/D hybrid with a broad host-range. AB - A recombinant interferon (IFN) hybrid has been found to have a broad host-range of activity in an antiviral assay (plaque reduction of vesicular stomatitis virus) and also high efficacy as an antiviral agent in at least 12 different animal cell species. The IFN hybrid consists of amino acids 1 to 60 from HuIFN alpha B and amino acids 61 to 166 from HuIFN-alpha D. The profile of cross species activity of the IFN-alpha B/D hybrid has been compared with that of HuIFN alpha F, and of the parents HuIFN-alpha B and -alpha D. When both IFN-alpha B and -alpha D were active in a cell species, the hybrid IFN had comparable or better activity than the more active parental IFN. The hybrid shared a broad cross species activity with IFN-alpha D. However, the IFN-alpha B/D hybrid was 10-fold more active on human cells, 30-fold more active on rabbit cells, and 50-fold more active on mouse cells than IFN-alpha D. PMID- 3029316 TI - Presence of episomal and integrated human papillomavirus DNA sequences in cervical carcinoma. AB - Thirty surgical samples of squamous cell carcinoma of the cervix obtained from Chinese women were analysed for the presence of human papillomavirus (HPV) types 16 and 18 using Southern blot hybridization procedure. HPV16 was detected in 53% while HPV18 was found in only 6% of the samples analyzed. When compared with other reports, variation in the geographic distribution of these two HPV types in association with cervical carcinoma is noted. Thirty-seven and a half percent of the HPV16-positive samples contained this HPV type in episomal form and an equal number in cellular DNA-integrated form. The simultaneous presence of both episomal and integrated forms was found in the remaining 25% of the positive samples. The two HPV18-positive cases harbored only episomal viral genome and were not superinfected by HPV16. Analysis of the HPV16 integration samples showed that single integration events had probably occurred and some of the viral sequences had been lost on or subsequent to integration. PMID- 3029317 TI - Subclass reactivity to Epstein-Barr virus capsid antigen in primary and reactivated EBV infections. AB - A new method for analysis of virus-specific Immunoglobulin G (IgG) subclasses was developed using indirect immunofluorescence. Three hundred thirty-three serum samples from patients with different types of Epstein-Barr virus (EBV)-associated diseases and healthy controls were examined for subclass distribution to the virus capsid antigen (EBV VCA). EBV-VCA-expressing cell preparations were incubated with patient serum followed by monoclonal antibodies to human IgG1 through IgG4 and labelled anti-mouse IgG. Virus-specific IgG1 was found to be the dominant antibody. The titers for IgG1 and total Ig to EBV VCA correlated well. EBV VCA-specific IgG2 was not found. EBV VCA-specific IgG3 in a titer of greater than or equal to 10 was found in 33% of healthy seropositive donors, in 97% of patients with suspected reactivated EBV infection, and in 100% of symptomatic patients with suspected reactivated EBV infection. EBV VCA specific IgG3 occurred in 90% of placebo-treated compared to 30% in long-term acyclovir-treated bone marrow transplant recipients, indicating more frequent reactivations in the former group. IgG4 to VCA was infrequently found in seropositive persons. In serum samples from patients with nasopharyngeal carcinoma and high EBV VCA Ig and IgA titers, IgG4 to VCA was always present. Analysis of EBV VCA specific IgG subclasses seems to be valuable for the diagnosis of reactivated EBV infection. PMID- 3029319 TI - Radioimmunoassay for the detection of IgG antibodies to herpes simplex virus and its use as a prognostic indicator of HSV excretion in transplant recipients. AB - We report the development of a solid-phase radioimmunoassay for the detection of IgG antibodies to herpes simplex virus (HSV), using a mouse monoclonal antibody specific for the Fc portion of human IgG as the radiolabelled detecting antibody. The binding ratio to test antigen at a single serum dilution, 1:100, correlated significantly with the endpoint titre by radioimmunoassay and with neutralising antibody titre. When compared to neutralisation the radioimmunoassay had a sensitivity of 100% and a specificity of 93%. We believe that the anomalous results are not false positives but represent an increased sensitivity of the radioimmunoassay. We found, by either radioimmunoassay or neutralisation, that high levels of antibody prior to transplantation were associated with a significantly increased risk of HSV excretion post-transplantation in both renal and bone marrow transplant recipients. Thus the radioimmunoassay is a sensitive, specific, and rapid test that can be used as a prognostic indicator of HSV excretion in transplant recipients. PMID- 3029318 TI - Antiviral antibodies in the sera of homosexual men: correlation with their lifestyle and drug usage. AB - Healthy homosexual men between the ages of 21 and 65 years, from the Washington, DC (n = 162), and New York City (n = 89) areas, were studied for antibodies in the serum against cytomegalovirus (CMV), herpes simplex virus (HSV) types 1 and 2, and Epstein Barr virus (EBV) viral capsid antigen (VCA). CMV-specific antibodies were assayed by enzyme-linked immunosorbent assay (ELISA), anti-HSV-1 and -2 antibodies were measured by indirect hemagglutination (IHA), and antibodies to EBV VCA were measured by the immunofluorescence assay. Antibodies to human T lymphotrophic virus III (HTLV-III) were detected by ELISA and Western blot procedures. T lymphocytes were enumerated using OKT4 monoclonal antibody. Healthy male volunteer blood donors (n = 90) matched for age range and race proportions were used as controls. The percentage of seropositive individuals in the homosexual group was higher (90-98%) for all the viruses tested than in the control group (47-87%). Comparisons of the geometric mean titers, expressed as reciprocal serum dilutions, of seropositive individuals in homosexual (H) vs control (C) group were as follows: CMV-IgG (ELISA) H = 1:794, C = 1:68; HSV-1 (IHA) H = 1:248, C = 1:14; HSV-2 (IHA) H = 1:56, C = 1:17; EBV-VCA (IFA) H = 1:385, C = 1:131. The homosexual group also showed a higher frequency of individuals with elevated titers than the control group. The CMV IgM antibody was prevalent in 17.7% of the homosexual group and 5% of the control group; arithmetic means for ELISA values for CMV IgM were 0.207 for the homosexual group and 0.05 for the control group. In the homosexual group, the anti-CMV antibody titers increased with age (P = 0.01) and with numbers of sex partners (P = 0.06). Both anti-HSV-1 and anti-HSV-2 antibodies correlated with the number of sex partners (P = 0.04 and P = 0.05, respectively). Neither age nor partner number correlated with response to EBV, and no particular sex act was related to the EBV VCA titer level. HTLV-III seropositivity was associated with higher herpes virus group antibody titers, probably because of life style cofactors. Among the HTLV III-seropositive subjects, those with less than or equal to 400 T-helper lymphocytes/mm3 had lower antibody titers than those with greater than 400 T helper lymphocytes/mm3 counts, suggesting an impaired immune response secondary to immunosuppression. PMID- 3029320 TI - Detection of viral antigens in cerebrospinal fluid of patients with herpes simplex virus encephalitis. AB - Thirty-two cerebrospinal fluid (CSF) samples from eighteen patients with confirmed herpes simplex encephalitis (HSE) were assayed by an indirect enzyme linked immunosorbent assay (ELISA) for the presence of viral antigens. The results are expressed as an antigen ratio distinguishing between herpes simplex virus (HSV) antigens containing samples and negative samples. Judged by this criterion a positive result was obtained in 33% of the patients. Overall, 25% of the CSF samples from HSE patients were positive. In one out of 33 control patients with other neurological disorders a positive antigen ratio was found. Two or more CSF samples were available from eleven patients. In six of these, the second or later samples showed a decreased antigen ratio when compared to the first CSF sample. An increase of the anti-HSV antibody titer was seen in the CSF of five of these six patients. Five out of six patients with a decreasing antigen ratio had an unfavorable outcome of their encephalitis, while a favorable outcome was seen in four of the five patients with an increasing or steady antigen ratio. A decrease of the antigen ratio in the course of HSE can be explained by the presence of immune complexes in CSF and may indicate a poor prognosis. PMID- 3029322 TI - Detection of adenovirus types 40 and 41 in stool specimens by immune electron microscopy. AB - An immune electron microscope (IEM) test was developed that allowed the direct detection of adenovirus type 40 (ad 40) or ad 41 in stools specimens. The polyclonal rabbit antisera used differentiated ad 40 and 41 from other ad serotypes but not from each other. The method was evaluated in a 13 month prospective study of stools from children with gastroenteritis. Seventy-two specimens found to contain ad by conventional electron microscope screening were retested by IEM. Results were typically obtained within 2 hr and showed that 55 (76%) viruses typed as ad 40/41. No ads were recovered from conventional virus isolation attempts on these specimens. Additionally, 39 of these 55 viruses were tested by restriction endonuclease analysis (REA) after growth in 293 cells, and results showed that all produced digest patterns typical of ad 40 (seven cases) or ad 41 (32 cases). Twenty-four percent (17/72) of viruses could not be typed by IEM; 9/17 (53%) yielded ads [ad 1 (1), ad 2 (4), ad 5 (1), ad 6 (1), ad 7 (2)] in routine culture, whereas REA identified the other eight as ad 2 (6), ad 1 (1), and ad 41 (1). The concordance between IEM and the reference methods was therefore 100% specificity and 97.5% sensitivity. The method described allows the clinically useful diagnosis of ad 40/41 infection to be rapidly made and will be a particularly useful technique in laboratories screening faeces by electron microscopy. PMID- 3029321 TI - Herpes simplex-specific IgG subclass response in herpetic keratitis. AB - The measurement of the local IgG response in ocular Herpes simplex virus infection presents particular problems due to the difficulty in obtaining sufficient tear samples and the possible transudation of IgG from the serum to the inflamed eye. Using specific monoclonal antibodies to Human IgG subclasses in an enzyme-linked immunoabsorbant assay (ELISA) the local IgG antibody response in Herpes simplex keratitis was analysed. All serum samples from patients and controls contained quantifiable levels of HSV specific IgG1 and IgG4 antibody. Comparison of serum antibody levels with tear levels for patients showed that HSV specific IgG1 serum concentrations were 16.1 fold or more higher than in tears, whereas IgG4 concentrations were only 6.5 fold higher in serum than in tears. This difference was not apparent in the control group. Radioimmunoprecipitation assay of 35S-methionine labelled HSV antigens revealed only minor differences in the protein profiles produced by immunoprecipitation using serum or tear antibody. These results suggest a role for IgG4 antibodies in mucosal immunity in the eye as has been suggested for the mucosal surface of the lung. PMID- 3029323 TI - Reduced phosphoinositide concentrations in anterior temporal cortex of Alzheimer diseased brains. AB - Samples of brain anterior temporal cortex from 17 patients with Alzheimer's disease and 18 age-matched controls have been analysed for myo-inositol and the three phosphoinositides. There was significantly less phosphatidylinositol in the Alzheimer samples (1.36 mumol/g wet weight) than in the controls (2.28 mumol/g). The concentrations of phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate were also lower in Alzheimer cortex, but differences from the control group were not statistically significant. Free myo-inositol concentrations were 5.11 mumol/g (Alzheimer) and 4.44 mumol/g (control) and again the difference was not significant. The lack of phosphoinositides in Alzheimer temporal cortex may impair receptor function. PMID- 3029324 TI - Morphine and enkephalins potently inhibit [3H]noradrenaline release from rat brain cortex synaptosomes: further evidence for a presynaptic localization of mu opioid receptors. AB - Synaptosomes prepared from rat cerebral cortex and labeled with [3H]noradrenaline (NA) were superfused with calcium-free Krebs-Ringer-bicarbonate medium and exposed to 10 mM K+ plus 0.1 mM Ca2+ so that [3H]NA release was induced. 6,7 Dihydroxy-N,N-dimethyl-2-aminotetralin (TL-99) strongly inhibited synaptosomal K+ induced [3H]NA release (EC50 = 5-10 nM) by activating alpha 2-adrenoceptors. Release was also inhibited (maximally by 40-50%) by morphine (EC50 = 5-10 nM), [Leu5]enkephalin (EC50 = approximately 300 nM), [D-Ala2,D-Leu5]enkephalin (DADLE), and Tyr-D-Ala-Gly-(NMe)Phe-Gly-ol (DAGO) (EC50 values = approximately 30 nM). In contrast to the mu-selective opioid receptor agonists morphine and DAGO, the highly delta-selective agonist [D-Pen2,D-Pen5]enkephalin (1 microM) did not affect [3H]-NA release. Furthermore, the inhibitory effect of DADLE, an agonist with affinity for both delta- and mu-opioid receptors, was antagonized by low concentrations of naloxone. The findings strongly support the view that, like alpha 2-adrenoceptors, mu-opioid receptors mediating inhibition of NA release in the rat cerebral cortex are localized on noradrenergic nerve terminals. PMID- 3029325 TI - Protein phosphorylation in rat pineal gland and its regulation in supersensitive and subsensitive states. AB - The phosphorylation of specific proteins in pineal homogenate was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. Cyclic AMP had the capacity to stimulate in a dose-dependent manner the incorporation of 32P in protein bands of apparent molecular weights 59K, 56K, and 35K with a maximal effect at 1 microM. On the other hand, calcium alone did not induce a marked increase in 32P incorporation with the exception of a dose dependent phosphorylation of a 46K protein with a peak effect at 0.2 mM calcium concentration. The addition of exogenous calmodulin enhanced 32P incorporation in proteins migrating in the 62K and 52K regions, an effect that was antagonized by the calmodulin inhibitor trifluoperazine. However, also under these conditions, the stimulation of pineal protein phosphorylation was rather weak compared to that observed in other brain areas. In an attempt to investigate the functional changes of these biochemical processes during environmental lighting and adrenergic stimulation, it was found that the administration of (-)-isoproterenol (5 mg/kg, s.c.), a beta-receptor agonist, induced a clear-cut enhancement of 32P incorporation into the cyclic AMP-sensitive 59K and 56K proteins only in animals exposed for 18 h to the light, whereas it was almost ineffective in those kept in the dark for the same period. This effect was antagonized by (-)-propranolol pretreatment (20 mg/kg), suggesting that the changes in cyclic AMP-dependent protein phosphorylation observed in supersensitive pineals may represent a beta receptor mediated process.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3029326 TI - Phylogenetic comparison of the photoaffinity-labeled benzodiazepine receptor subunits. AB - The late evolutionary appearance of the benzodiazepine receptor (BZR) allows an experimental approach for evaluation of the qualitative development of its subunits. Photoaffinity labeling of brain membranes with [3H]flunitrazepam followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography offers a suitable method for tracing the qualitative evolution of the BZR. A systematic comparison of the subunit patterns in fishes, amphibians, reptiles, birds, and mammals revealed that the subunit of 53K is phylogenetically the oldest photoaffinity labeled subunit; whereas it is the only band present in the lungfish and most amphibians, additional bands are apparent in higher tetrapods. In fishes, the evolution of the BZR subunits leads to the loss of the 53K subunit. KD values are discussed in relation to specific subunit patterns. Possible explanations for the observed variation of the subunits are discussed, with special emphasis placed on the possible evolution by gene duplication and subsequent divergence. PMID- 3029327 TI - Comparison of tryptic peptides of benzodiazepine binding proteins photolabeled with [3H]flunitrazepam or [3H]Ro 15-4513. AB - When rat brain membranes were incubated with the benzodiazepine agonist [3H]flunitrazepam or the partial inverse benzodiazepine agonist [3H]Ro 15-4513 in the presence of ultraviolet light one protein (P51) was specifically and irreversibly labeled in cerebellum and at least two proteins (P51 and P55) were labeled in hippocampus. After digestion of the membranes with trypsin, protein P51 was degraded into several peptides. When P51 was photolabeled with [3H]Ro 15 4513, four peptides with apparent molecular weights of 39,000, 29,000, 21,000, and 17,000 were observed. When P51 was labeled with [3H]flunitrazepam, only two peptides with apparent molecular weights of 39,000 and 25,000 were obtained. Protein P55 was only partially degraded by trypsin, and whether it was labeled with [3H]flunitrazepam or [3H]Ro 15-4513 it yielded the same two proteolytic peptides with apparent molecular weights of 42,000 and 45,000. These results support the existence of at least two different benzodiazepine receptor subtypes associated with proteins P51 and P55. The different receptors seem to be differentially protected against treatment with trypsin. In addition, these results indicate that in the benzodiazepine receptor subtype associated with P51 benzodiazepine agonists and partial inverse benzodiazepine agonists irreversibly bind to different parts of the molecule. PMID- 3029328 TI - Guanine nucleotide and cation regulation of mu, delta, and kappa opioid receptor binding: evidence for differential postnatal development in rat brain. AB - A study of the onset of cation and guanine nucleotide regulation of delta, mu, and kappa rat brain opioid receptors during postnatal development was undertaken. Site-specific binding assays were utilized for each receptor type and the effects of 0.5 mM MnCl2, 100 mM NaCl, and/or 50 microM guanosine-5'-(beta, gamma-imido) triphosphate [Gpp(NH)p] were assessed. The most pronounced changes of opioid binding were seen in the presence of Mn2+. In adults, agonist binding to delta sites was stimulated by Mn2+, whereas that to mu sites was not affected and kappa binding was inhibited. The postnatal development of Mn2+ regulation for the three receptor subtypes was distinctly different. The largest effects were seen on delta sites detected in the early neonatal period, Mn2+ eliciting a 68% stimulation of binding over controls at day 1. Significant inhibition of kappa site binding by Mn2+ was detected only after the third postnatal week. Mn2+ caused a significant reversal of Gpp(NH)p inhibition of delta binding in the early neonatal period, exceeding that in the absence of regulators. Inhibition of mu and delta receptor binding by Na+ was greater, and the Mn2+ reversal of this effect was smaller, in the first 2 postnatal weeks than in adults. Gpp(NH)p + Na+ regulation did not change appreciably during the postnatal period. However, Mn2+ reversal of the considerable inhibition elicited by the combination of Na+ and Gpp(HN)p was developmental time-dependent. The data are discussed in terms of multiple sites of interaction for guanine nucleotides and cations.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3029329 TI - Solubilization and characterization of the nitrendipine receptor in rat brain. AB - The nitrendipine receptor associated with the voltage-dependent calcium channel in rat brain was solubilized by detergent extraction and sonication. The detergent solution used for extraction consisted of 10 mM 3-[(3 cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS), 0.25% (wt/vol) polyoxyethylene 20 cetyl ether (Brij 58), and 0.025% (wt/vol) polyoxyethylene 17 cetyl stearyl ether (Lubrol WX) in the presence of 30% (wt/vol) glycerol as a stabilizer. The molecular weight of the receptor was estimated to be 1,800K by Sephacryl S-500 gel filtration and 800K by sucrose density gradient sedimentation. The equilibrium dissociation constant of [3H]nitrendipine to the solubilized receptors was 5.6 nM, which is approximately 10 times that of the membrane-bound receptor. The binding of nitrendipine to the receptor was inhibited noncompetitively by the structurally unrelated calcium channel inhibitors verapamil and prenylamine; their concentrations for 50% inhibition were both 1.0 X 10(-7) M, and they caused maximal inhibitions of 70 and 100%, respectively. PMID- 3029330 TI - Changes in beta-adrenergic receptor subtypes in Alzheimer-type dementia. AB - Using ligand binding techniques, we studied beta-adrenergic receptor subtypes in brains obtained at autopsy from seven histologically normal controls and seven histopathologically verified cases with Alzheimer-type dementia (ATD). Inhibition of [3H]dihydroalprenolol [( 3H]DHA) binding by the selective beta 1 antagonist, metoprolol, results in nonlinear Hofstee plots, suggesting the presence of the two receptor subtypes in the human brain. The calculated ratios of beta 1/beta 2 adrenergic receptors in control brains are as follows: frontal cortex, 49:51; temporal cortex, 31:69; hippocampus, 66:34; thalamus, 23:77; putamen, 70:30; caudate, 48:52; nucleus basalis of Meynert (NbM), 43:57; cerebellar hemisphere, 25:75. Compared with the controls, total concentrations of beta-adrenergic receptors were significantly reduced only in the thalamus of the ATD brains. beta 1-Adrenergic receptor concentrations were significantly reduced in the hippocampus and increased in the NbM and cerebellar hemisphere, whereas beta 2 adrenergic receptor concentrations were significantly reduced in the thalamus, NbM, and cerebellar hemisphere and increased in the hippocampus and putamen of the ATD brains. These results suggest that beta 1- and beta 2-adrenergic receptors are present in the human brain and that there are significant changes in both receptor subtypes in selected brain regions in patients with ATD. PMID- 3029331 TI - Delta sleep-inducing peptide modulates the stimulation of rat pineal N acetyltransferase activity by involving the alpha 1-adrenergic receptor. AB - Delta sleep-inducing peptide (DSIP) has been isolated and characterized by its capacity to enhance delta sleep in rabbits. Up to now, sleep was the main target of DSIP research, but different extra-sleep effects of the peptide have been reported as well. Several mechanisms of action have been proposed, though no convincing evidence for any of them has been obtained so far. We recently detected that DSIP reduced the nocturnal increase of N-acetyltransferase (NAT) activity in rat pineal in a dose-dependent manner. The activity of this enzyme is known to be induced by adrenergic agonists and several studies have suggested that stimulation of alpha 1-adrenergic receptors potentiates the "basic" effect of beta-receptors. DSIP in the range between 20 and 300 nM significantly enhanced NAT activity induced by 10(-6) M norepinephrine in vitro, and a similar effect was observed with 2nMP-DSIP, a phosphorylated analog. Incubation with prazosin eliminated the enhancement, whereas propranolol reduced norepinephrine stimulation that was still increased by P-DSIP and probably DSIP. It was concluded that the sleep-peptide and its analog modulate the alpha 1-adrenergic receptor of rat pineal in its response to adrenergic agonists. The same mechanism may also be responsible for other biological activities of DSIP such as sleep induction and stress-tolerance. PMID- 3029332 TI - Postmortem changes in rat brain: studies on membrane-bound enzymes and receptors. AB - The relationship between the stability of potential neurochemical markers and autolysis time was studied at 4 degrees C and 25 degrees C using postmortem brain samples from two rat strains. In general, qualitatively similar results were obtained with either N/Nih or Sprague-Dawley rats; however, quantitative differences were often observed, particularly in regard to benzodiazepine receptor changes. For every enzyme activity or binding property examined, no significant change was found when brains were kept at 4 degrees C for up to 72 h prior to freezing at -70 degrees C. Na,K-ATPase and low-affinity Ca-ATPase activities were also stable in brains kept at 25 degrees C for up to 72 h. Mg ATPase activity was reduced in brains kept at 25 degrees C for 24 and 48 h. [3H]Guanidinoethylmercaptosuccinic acid [( 3H]GEMSA) binding to enkephalin convertase in the cytosol was not significantly changed in brains kept at 25 degrees C; however, a small increase was seen for [3H]GEMSA binding to the membrane fraction at 24, but not 48 and 72 h postmortem. [3H]Quinuclidinyl benzilate [( 3H]QNB) binding to muscarinic cholinergic receptors decreased in brains kept at 25 degrees C for 72 h. Opioid receptor binding also decreased in brains kept at 25 degrees C. Using [3H]2-D-alanine-5-D-leucine enkephalin to label delta opioid receptors, a statistically significant decrease in binding was observed as early as 6 h postmortem, and was completely abolished after 72 h at 25 degrees C. In contrast, [3H]naloxone binding was unchanged after 24 h at 25 degrees C, but was decreased after 48 and 72 h.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3029333 TI - Receptor activation and inositol lipid hydrolysis in neural tissues. PMID- 3029334 TI - Lymphomatous meningitis appearing as Garin-Bujadoux-Bannwarth meningopolyneuritis. PMID- 3029336 TI - Peripheral neuropathy in mitochondrial disease. AB - Clinical, electrophysiological, histological and biochemical studies of two patients with mitochondrial disease revealed a moderately advanced axonal neuropathy with mitochondrial paracrystalline inclusions in Schwann cells, fibroblasts and muscle fibers. In addition there was a myopathy, and the activity of muscle cytochrome c oxidase was diminished by more than 50%. There were electrophysiological signs of myopathy, neuropathy and failure of excitation contraction coupling in both patients. The partial enzyme deficiency raises some questions as to its pathogenetic role in these neuromyopathies. PMID- 3029335 TI - Demyelinating neuropathy associated with hepatitis B virus infection. Detection of immune complexes composed of hepatitis B virus surface antigen. AB - We observed 4 patients with the Guillain-Barre syndrome (GBS) type of polyneuropathy and one patient with chronic relapsing polyneuropathy associated with hepatitis B virus infection, and examined the sera, cerebrospinal fluid (CSF) and sural nerve specimens from the patients in search of the pathogenetic factors involved. It was demonstrated that hepatitis B surface antigen(HBsAg) immune complexes were significantly increased in both the sera and the CSF of the 4 patients with GBS. The serum levels of immune complexes were also closely related to the clinical status of these patients. In all patients, HBsAg-positive labelling of immunofluorescence was found around the endoneural small blood vessels and in the endoneurium. Electron-dense deposits, suggestive of immune complexes composed of hepatitis B virus, were demonstrated in the endoneurium of the patient with chronic relapsing polyneuropathy. These results suggest that HBsAg immune complexes may be of importance in the etiology of GBS or chronic relapsing polyneuropathy associated with hepatitis B virus infection. PMID- 3029337 TI - Peripheral neuropathy associated with experimental plasma cell neoplasm in the mouse. AB - In order to investigate the role of paraproteins in the production of neuropathy we experimentally induced monoclonal immunoglobulin-producing tumours into 32 BALB/c mice by injecting mineral oil or pristane intraperitoneally. In 11 mice morphologic and histometric studies of the sciatic nerve revealed the presence of neuropathy. Immunohistochemical studies did not demonstrate a significant amount of immunoglobulins and light chains in the endoneurium. The advantages and limits of this experimental model are discussed. PMID- 3029338 TI - Hirano body filaments contain actin and actin-associated proteins. AB - Hirano bodies are eosinophilic, rod-shaped intraneuronal inclusions whose frequency increases with age and with Alzheimer's disease. To investigate their composition and possible relationship to the neuronal cytoskeleton, we employed immunocytochemistry and immunoelectronmicroscopy by using antisera to cytoskeletal proteins. The presence of actin, alpha-actinin, vinculin and tropomyosin was demonstrated diffusely throughout the Hirano body. The presence of these proteins supports the contention that Hirano bodies are derived from an abnormal organization of the neuronal cytoskeleton. The staining of Hirano bodies with fluorescent labelled phalloidin, a probe with a unique affinity for F-actin, indicates that the actin in Hirano bodies is in the F-state. Results of high voltage electron microscopy on 1.0 and 0.5 micron sections confirm the purely filamentous nature of Hirano bodies. These findings suggest that the mechanism of formation of Hirano bodies is different from that of the neurofibrillary tangle, another characteristic intraneuronal inclusion seen in Alzheimer patients. PMID- 3029340 TI - Evaluation of three biochemical markers for serially monitoring the therapy of small-cell lung cancer. AB - A number of biochemical markers have been proposed for monitoring the therapy of small-cell lung cancer (SCLC). This report reviews the experience at a single institution using three biochemical markers, alpha-1-acid glycoprotein (AGP), carcinoembryonic antigen (CEA), and lactate dehydrogenase (LDH), for serially monitoring the therapy of patients with SCLC. AGP measurements identified limited disease patients more frequently than LDH or CEA, and a combination of markers (AGP and LDH) improves the accuracy of correctly classifying patients with active disease. Each of the markers correctly tracked the clinical response to therapy in approximately two thirds of the subjects. The use of a combination of markers should be considered for monitoring the therapy of SCLC in future clinical trials. PMID- 3029339 TI - Phase II diaziquone-based chemotherapy trials in patients with anaplastic supratentorial astrocytic neoplasms. AB - We treated 103 patients with histologically confirmed anaplastic supratentorial astrocytic neoplasms with either diaziquone (AZQ) and carmustine (BCNU) or AZQ and procarbazine. There were 74 patients with glioblastoma multiforme (GBM) and 29 patients with anaplastic astrocytoma (AA). AZQ plus BCNU produced partial (PR) or unequivocal responses in seven of 32 (21.9%) patients with GBMs and three of ten (30%) patients with AAs. Two patients with GBMs (6.3%) and five patients with AAs (50%) showed stable disease (SD). AZQ plus procarbazine produced PRs or unequivocal responses in five of 42 (11.9%) patients with GBMs and nine of 19 (47.4%) patients with AAs. Eight patients with GBMs (19%) and one patient with an AA (5.2%) showed SD. In addition to histologic diagnosis, only the Karnofsky performance-status (KPS) rating independently influenced response and survival. Differences in response rates between the two regimens were not significant, although estimated median survival after adjusting for performance status was slightly better with AZQ plus BCNU than with AZQ plus procarbazine (P = .031). Neither age nor prior chemotherapy were significant independent risk factors. Toxicity was mild and primarily hematologic. We conclude that these AZQ-based regimens have activity in patients with recurrent anaplastic gliomas, but that they are not clearly superior to other agents in current use. The histologic diagnosis of GBM is associated with a significantly worse prognosis than AA, and we believe that this important distinction must be recognized in phase II as well as phase III trials. PMID- 3029341 TI - Forskolin's effect on transient K current in nudibranch neurons is not reproduced by cAMP. AB - Forskolin, a diterpene extracted from Coleus forskolii, stimulates the production of cAMP in a variety of cells and is potentially an important tool for studying the role of cAMP in the modulation of neuronal excitability. We studied the effects of forskolin on neurons of nudibranch molluscs and found that it caused characteristic, reversible changes in the amplitude and waveform of the transient K current, IA, and also activated an inward current similar to the cAMP-dependent inward current previously described in molluscan neurons. Forskolin altered the time course of IA activation and inactivation but did not affect the voltage dependence or the reversal potential of the current. IA normally inactivates exponentially, but in forskolin the time course of inactivation can be fit by the sum of 2 exponentials with an initial rate that is faster than the control and a final rate that is much slower. On depolarization in forskolin, IA begins to activate at the normal rate, but a slower component of activation is also seen. The changes in IA in the nudibranch cells were qualitatively different than the changes caused by forskolin in Aplysia bag cell neurons (Strong, 1984). Experiments were performed to determine whether these effects of forskolin require cAMP. Intracellular injection of cAMP, application of membrane-permeable analogs of cAMP, application of phosphodiesterase inhibitors, and intracellular injection of the active catalytic subunit of cAMP-dependent protein kinase did not affect the amplitude or waveform of IA. Also, the changes in IA that are caused by forskolin were not prevented or reversed by intracellular injection of an inhibitor of cAMP-dependent protein kinase. Cyclic AMP did, however, activate inward current at voltages near the resting potential. We conclude that the changes in IA and the activation of inward current represent separate affects of forskolin. The inward current appears to depend on an increase in intracellular cAMP, while the changes in IA do not. These experiments show that, in addition to activating adenylate cyclase, forskolin may have a separate direct affect on the transient K current. PMID- 3029342 TI - Structure and physiology of developing neuromuscular synapses in culture. AB - The structure and function of developing neuromuscular synapses in culture have been investigated. We used neuromuscular junctions formed by coculturing dissociated muscle cells and dissociated neurons obtained from Xenopus embryos. After recording nerve-evoked endplate potentials (e.p.p.s) and spontaneously occurring miniature endplate potentials (m.e.p.p.s) from a given junction, the same specimen was investigated for electron-microscopic histology. We surveyed almost the total area of the junctional region by making serial sections. Even in preparations cocultured for only a short time (4-11 hr), both e.p.p.s and m.e.p.p.s could be obtained. The junctional region of these early synapses revealed a simple structure. The presynaptic terminals contained smooth-surfaced clear vesicles, but there were no presynaptic specializations such as active zones. The width of the synaptic cleft was variable, with predominance of narrow regions (10-30 nm), and there was no basal lamina inside the cleft. When the coculture time was 1 d or longer, the junctional area started to show structural features resembling a mature neuromuscular synapse. In the presynaptic terminal there were active zones, consisting of the presynaptic density and an accumulation of vesicles near the density. In many junctions, the postsynaptic membrane showed densities and thickenings, with a widened synaptic cleft, that contained basal lamina. It is known that growth cones, prior to making neuromuscular junctions, can release the transmitter substance with a very long latency if stimulated repetitively. In contrast, e.p.p.s with short latencies can be evoked by single stimuli soon after the growth cones attach to muscle cells. However, our data did not reveal any structural changes to account for such functional changes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3029343 TI - Antidepressant drug-induced changes in brain beta-adrenergic receptors: subcellular studies. PMID- 3029344 TI - Preoperative parathyroid localization by superimposed iodine-131 toluidine blue and technetium-99m pertechnetate imaging. AB - A new parathyroid scintigraphic localization study by a dual radioisotope technique using radioiodinated toluidine blue (RTB) for the parathyroids and 99mTc for thyroid imaging is presented. A simple RTB labeling procedure achieving 99% tagging of the 131I-TB was used. The RTB was found to be a highly specific parathyroid radiotracer, consequently enabling superimposition of the delineated thyroid gland over the RTB avid parathyroid foci without a need for subtraction of the thyroid or vascular background. Forty-six patients with primary hyperparathyroidism underwent scintigraphic study prior to cervical (41 patients) or mediastinal (5 patients) exploration and 67 pathological parathyroid glands (34 adenomas and 33 hyperplasias) were excised. On follow-up, serum calcium level returned to normal in all patients. Correlation of the scintigraphic results with the surgical findings disclosed a sensitivity of 93%, with a specificity of 80% and an overall accuracy of 87%. This new simplified and specific RTB scintigraphic method justifies its use as a routine procedure for preoperative parathyroid scintigraphic localization in primary hyperparathyroidism. PMID- 3029345 TI - Effect of antidepressant and narcoleptic drugs on N-isopropyl p-iodoamphetamine biodistribution in animals. AB - N-isopropyl p-iodoamphetamine (IMP) demonstrates a high affinity for lung and brain during the first pass following intravenous injection. Its high brain affinity has been used to advantage for cerebral perfusion imaging, but the effects of drugs on IMP distribution could affect its utility. In this study, we determined the effects of the tricyclic antidepressant imipramine and the MAO inhibitors deprenyl and phenelzine on the biodistribution of IMP. We first determined the effect of loading dose and anesthesia on the biodistribution of IMP. In rats, biodistribution was not dependent on loading dose between 0.1 and 1.1 mg/kg. Anesthesia with thiopental and chloral hydrate depressed lung and brain IMP uptake. In rats, preloading doses of imipramine depressed lung uptake but did not result in increased brain IMP uptake; postloading doses of imipramine did not release IMP from the lung. In rabbits, simultaneous or postloading doses of imipramine resulted in release of IMP from the lung with an increase in brain activity. Both mixed A and B MAO inhibitors (phenelzine) and B selective MAO inhibitors (deprenyl) did not affect IMP distribution in rats. Based on the action of imipramine on IMP uptake and clearance in the lung, we postulate that IMP uptake and metabolism within the lung is related to the mixed function oxidase (MFO) system. As the lung is rich in the MFO system in humans, we would also predict from this study that IMP distribution in patients under antidepressant therapy would not be affected by either tricyclic or MAO inhibitor agents apart from the effect of these drugs on cerebral perfusion. PMID- 3029346 TI - Test anxiety in master's students: a comparative study. AB - Evaluation events are major problems for graduate students in nursing. The purpose of this study was to determine the correlation of test anxiety before and cognitive interferences during a comprehensive examination. The sample (N = 54) consisted of two groups of students (Group I, N = 30 and Group II, N = 24) in a master's program at a state university. Data were collected using three questionnaires: To measure general test anxiety, the Test Anxiety Questionnaire (TAQ) by Sarason was used; the Pre-Examination Questionnaire (PEQ) by Morris, Davis, and Hutchings (1981) was used to measure pretest anxiety; and to identify cognitive interferences during the examination, the Cognitive Interference Questionnaire (CIQ) by Sarason and Stoops (1976) was used. Findings revealed positive correlations between the two groups of students' general test anxiety and their cognitive interferences about evaluative outcome of the examination. Positive correlations were found also between the two groups' general test anxiety and their pretest anxiety immediately prior to writing the examination. No significant relationships were found between the student's performance rankings on the examination and their test anxiety. PMID- 3029347 TI - Learning of psychomotor skills: laboratory versus patient care setting. PMID- 3029348 TI - Factors in academia and service that influence baccalaureate graduates' membership in the American Nurses' Association. AB - The intent of this study was to explore relationships among models of influence, association experiences, attitudes toward the American Nurses' Association (ANA), attitudes toward professionalism, and first-year baccalaureate nurse graduates joining the American Nurses' Association. One hundred sixty-three baccalaureate nurses from programs in three northeastern states returned a mailed questionnaire. Analysis of the data showed that graduates identified deans and faculty as more favorably disposed toward the ANA as compared to supervisors, head nurses, and peers. Faculty were identified more often as influencing subjects to join the ANA. Respondents saw the ANA as valuable for the profession, representing nursing, and promoting standards but they generally did not hold membership. Professionalism content in the curriculum had no relationship to scores on either of the two scales or to ANA membership. Having had ANA content was positively related to seeing the ANA as promoter of human rights and as a voice for nursing. There were significant differences between ANA members and non members and use of the professional organization as a major referent and seeing the ANA as an accreditor. It was concluded that faculty and deans were positive models of influence for new graduates regarding the Association but did not affect membership for those graduates. Further study of curriculum content and professional role model behavior in the work setting is urgently needed. PMID- 3029349 TI - Computer-managed instruction: an alternative teaching strategy. AB - Computer-managed instruction is an instructional strategy whereby the computer is used to provide learning objectives, learning resources, and assessment of learner performance. Computer-managed instruction (CMI) aids the instructor in instructional management without actually doing the teaching. Central CMI themes discussed in the literature are individualization, behavioral objectives, and educational technology. The main objective of this study was to compare the outcomes of two teaching strategies: CMI versus the traditional lecture method. The learning objectives were based on specified theoretical content from a Health Assessment course for baccalaureate nursing students. The design of the study was quasi-experimental incorporating two experimental treatments applied to two groups on two occasions. Data analysis addresses differences between groups using CMI and the traditional lecture method. The variables examined were the cognitive performance of learners, the learner's attitude toward the instructional strategy, the learner's retention of knowledge, the time involved in mastering the learning objectives, and the relationship between learner characteristics and the effectiveness of the instructional strategy. No significant mean difference (p less than 0.05) was found between groups on cognitive performance as measured by written and practical examination scores. For these first-year baccalaureate nursing students, CMI did not prove to be a positive instructional method as assessed by the Attitude Questionnaire. The majority of students preferred a combination of instructional methods. There was no significant difference between groups in the time spent meeting the learning objectives by either teaching strategy. The findings suggested that these students preferred learning strategies that are traditional in nature and teacher directed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3029350 TI - The use of the seminar to teach group process and group leadership with registered nurse students at the baccalaureate level. AB - Group process and group leadership "are areas in which definite skills can and must be learned; we are not all intuitively capable of working effectively with groups" (Sampson & Marthas, 1977, p. 18). A knowledge of effective group process and leadership ability in working with groups are essential for RN baccalaureate graduates who will be working with groups in a variety of settings in professional nursing practice. Through the use of the seminar as a teaching method, students learn the principles of group process and group leadership, function as a member of a group, and have the opportunity to practice leadership skills through leading seminar discussions. All of these experiences are vital components of baccalaureate nursing education for RN students. PMID- 3029352 TI - Evaluating borderline students. AB - While it was difficult for our faculty to face the disparity which arose in deciding whether or not to pass Anne, many important points were raised which strengthened the evaluation process particularly in the case of the borderline student. Newer faculty members related uneasiness in being specific and directive with "borderline" students. Experienced faculty offered suggestions on overcoming this fear and finding support among peers. Open discussion provided a forum for the exchange of ideas and philosophies. Though no definitive solutions were uncovered we judged this workshop to be a valuable learning experience with broad appeal across a very diverse faculty. PMID- 3029351 TI - The objective structured clinical examination: an alternative approach to assessing student clinical performance. PMID- 3029353 TI - The impact of nursing education on ethical/moral decision making. AB - A study of 227 baccalaureate and 111 master's nursing students was conducted to determine the influence of the level of formal education on three selected factors: ethical/moral reasoning, attribution of responsibility, and ethical/moral dilemma resolution. Moral development theory and Heider's attribution of responsibility construct provided the theoretical framework. A comparison of the overall index of ethical/moral reasoning showed that graduate students reasoned at a higher level than undergraduate students. The amount of attribution of responsibility assigned and the dilemma resolution score did not differ for the two groups. The results of this study suggest that undergraduate and graduate nursing programs must place more emphasis on identifying dilemmas, increasing ethical/moral reasoning levels and attributing responsibility in a justifiable manner. Nurse researchers must continue to study how nurses respond in dilemma situations and how personal characteristics, factors in the environment, education, and the assignment of responsibility affect nurses' ability to resolve ethical/moral dilemmas. PMID- 3029355 TI - Nutrient homeostasis: long-term interrelations between pancreatic hormones, blood glucose and dietary wheat bran in men. AB - Postprandial plasma somatostatin (SLI), pancreatic glucagon, insulin (IRI) and blood glucose (BG) were measured at 0, 30, 60, 120 and 180 min after a test meal in 10 healthy men before and after 1, 3 and 7 wk of ingestion of a balanced diet supplemented daily with 20 g wheat bran. BG response to the test meal was significantly and consistently lower after the first week of consuming bran, with a maximum drop after 3 wk. After 1 wk of bran, when compared with the prebran values, SLI secretion was decreased, and glucagon response was significantly enhanced at 120 and 180 min after the meal. IRI secretion did not change significantly until the third week. After consuming bran for 7 wk, postprandial SLI concentrations returned to prebran values, and glucagon levels were not significantly different from those of the first meal. IRI during the fourth test meal (after 7 wk of bran consumption) was significantly higher than after the third meal (after 3 wk of bran ingestion), at 60 min and at 120 min. BG levels remained low. Chronic bran ingestion was therefore associated with an improved glucose tolerance. Its effects on pancreatic hormones varied with time for each hormone, and somatostatin was the only one to return to the prebran values. PMID- 3029354 TI - Digestibility of boiled and oven-dried cassava in infants and small children. AB - Boiled fresh cassava or oven-dried cassava flour provided 50% of the energy and less than 10% of the protein in diets of convalescent malnourished infants; casein was added to complete 8% of energy as protein, and vegetable oil (soybean cottonseed) was added to make 20% of energy as fat. Despite high wet weights of feces (172 +/- 42 and 214 +/- 41 g/d), their dry weights (20 +/- 3 and 22 +/- 2 g/d) and their energy (6.9 +/- 0.7 and 7.6 +/- 0.5% of intake) and nitrogen (17 +/- 3 and 20 +/- 3% of intake) contents were low, and their fat content (4.4 +/- 1.3 and 5.2 +/- 1.2% of intake) was very low. Cassava is a surprisingly effective source of energy which interferes little or not at all with digestion of added protein and fat in weaning diets. For its safe use, it is important that home or industrial processing almost completely eliminate its potential hydrocyanic acid content, and that a good quality protein supplement be consumed regularly in nutritionally adequate amounts. PMID- 3029357 TI - The air-powder dental abrasive unit--an evaluation using a model system. AB - Air-powder abrasive systems are used in dentistry to remove dental plaque, calculus and stain from the surfaces of teeth. A model system consisting of a thin layer (14 micron) of vacuum-deposited aluminium was used to demonstrate the effectiveness of such a system. Photomicrographic analysis showed two distinct areas of removal: an inner area where removal of aluminium was complete, surrounded by an area where removal was incomplete. With the aid of a relocation pin attached to the nozzle of the instrument it was demonstrated that nozzle target distance, the air pressure input, the flow of water and the time of operation were all factors which affected the clinical efficiency of the instrument. PMID- 3029356 TI - Mineral balances of men and women consuming high fiber diets with complex or simple carbohydrate. AB - Calcium, copper, iron, magnesium, manganese, and zinc balances of 20 men, 19 premenopausal women, and 12 postmenopausal women were determined while they consumed self-selected diets or high fiber diets with either complex or simple carbohydrate. Self-selected intakes of calcium, magnesium, copper and zinc were generally below RDA levels. Iron intakes of premenopausal women were less than two-thirds RDA. Balances during the self-selected intake period of calcium in men and women and of magnesium and zinc in women were negative, but lower-than-usual intakes could have contributed to this. High fiber diets did not adversely affect zinc, copper, manganese or iron balances. Calcium and magnesium balances were generally negative, especially in women, even though intakes were adequate. Excretion of copper was increased in premenopausal women consuming the simple carbohydrate diet. The levels of magnesium and especially calcium needed to maintain balance when consuming a high fiber diet may be above present RDA levels. PMID- 3029358 TI - Porous hydroxyapatite as a bone graft substitute in diaphyseal defects: a histometric study. AB - Porous hydroxyapatite (IP200), formed by conversion of the Poritidae porites exoskeleton, has pores averaging 230 microns and pore interconnections averaging 190 microns in diameter. In the distal radial diaphyses of 14 dogs, bilateral 7.5 X 20 mm cortical windows were created and fitted with 5 X 7.5 X 20 mm blocks of IP200 implants and iliac autografts. Both implanted and contralateral grafted radius specimens were retrieved at 3, 6, 12, 24, and 48 months. Unstained undecalcified sections were examined by microradiography and UV epi-illumination. Stained undecalcified sections were examined by light microscopy and quantitated by histometric methods. Implant specimens demonstrated good union and bone ingrowth at all time intervals. The implant specimens were composed of (mean +/- SE) 10.6% +/- 1.0% soft tissue, 51.2% +/- 1.3% bone, and 38.2% +/- 1.0% IP200. The graft specimens showed good union with little apparent ingrowth at 3 months, followed by progressive appositional closure of cancellous spaces. The graft specimens contained 21.9% +/- 0.9% bone at 3 months with increases at each time interval to 73.1% +/- 8.7% at 48 months. The volume fraction and mean width of IP200 did not change with time, confirming the absence of implant biodegradation. The volume fraction and mean width of bone remained stable in the implant but increased in the graft specimens, corresponding to graft neocortex formation. It is concluded that implants initially filled in with bone while grafts initially replaced much of their spongiosa and subsequently filled in with bone. Histometry of untreated defects and measurement of mechanical properties are suggested for further study. PMID- 3029359 TI - [Characteristics of antibody production in tonsillar B-cells stimulated by Epstein-Barr virus (EBV) infection]. PMID- 3029360 TI - [Cystic lesions of the parotid gland--a clinicopathological study]. PMID- 3029361 TI - Viral supraglottitis. PMID- 3029362 TI - Maternal and fetal endocrine stress response at vaginal delivery with and without an epidural block. AB - Maternal and fetal stress response at vaginal delivery were studied in 19 normal parturients at term. Ten patients to whom an epidural block (group EA) had been administered were compared with 9 patients (group NEA) who used only nitrous oxide for pain relief. Plasma concentrations of ACTH, cortisol, 17-alpha hydroxyprogesterone, blood glucose and catecholamines were measured in maternal and umbilical vein blood at delivery and in maternal vein blood 30 minutes after delivery. At delivery maternal plasma concentrations of ACTH, cortisol and catecholamines were lower in the EA group compared with in the NEA group. There were no differences in umbilical plasma concentrations of the studied stress variables between the two groups. A linear relation was demonstrated between maternal and umbilical vein cortisol concentration. In both the EA and NEA group a significant fall in ACTH, 17-alpha-hydroxyprogesterone and catecholamine concentrations were demonstrated 30 minutes after delivery, whereas cortisol and blood glucose were virtually unchanged. It was found that epidural anesthesia reduced the maternal stress hormones at delivery but seemed to have little or no effect on the fetal endocrine stress hormones. PMID- 3029363 TI - Quantitation and optimization of enzymatic and mechanical procedures to procedures to produce high-yield single cell suspensions from human gingiva. PMID- 3029364 TI - Human gingival proteases. I: Extraction and preliminary characterization of trypsin-like and elastase-like enzymes. PMID- 3029365 TI - The presence of an inhibitor of human skin collagenase in the roots of healthy and periodontally diseased teeth. PMID- 3029366 TI - Histologic evaluation of periodontal implants in a biologically "closed" model. AB - To study the biologic response, three commercial calcium phosphate implant materials (Calcitite, Periograf, Synthograft) were implanted in cuspid root "windows" in four beagle dogs. No implant material was placed in the fourth cuspid window which served as a control. Following mucoperiosteal flap elevation, windows were chiseled in bone to the root surface which allowed implant particles to contact bone, fibrous connective tissue, cementum and dentin without exposure to the oral environment. Animals were sacrificed after 1, 3, 5 and 6 months. Histologically, all implant materials were well tolerated. At 1 month, implant particles were surrounded by fibrous tissue. Fibrous tissues filled the control defect. At 3 months, implant sites exhibited partial bony repair with connective tissue surrounding implant particles. A ring of osteoid surrounded Synthograft particles. Control defect repair was complete. At 5 and 6 months, implant sites exhibited advanced, though incomplete, bony repair. New bone encompassed the Synthograft particles. It was concluded: Control sites healed most rapidly. Calcitite and Periograft were well tolerated space occupiers. Synthograft was a nidus for bone deposition. PMID- 3029367 TI - Pharmacological actions of leukotrienes C4, D4 and E4 on guinea pig isolated taenia caecum as a preparation to study leukotrienes. AB - The contractile effects of leukotrienes (LTs) C4, D4 and E4 were studied in guinea pigs isolated taenia caecum. LTs C4, D4 and E4 contracted taenia caecum at low concentrations in a concentration-dependent manner. The response to LTs was not influenced by a gamma-glutamyl transpeptidase inhibitor (L-serine borate), aminopeptidase inhibitor (L-cysteine) or cyclooxygenase inhibitor (flurbiprofen), suggesting that LTC4 and LTD4 were not metabolized or metabolized only slightly in the guinea pig isolated taenia caecum, and that each of the LTs showed the direct contractile action. Furthermore, FPL55712 antagonized the response to LTD4 and LTC4 competitively but not that to LTE4. These results suggested that the guinea pig isolated taenia caecum is useful preparation to study the pharmacological actions of LTs. PMID- 3029368 TI - Influence of follicular health on the steroidogenic and morphological characteristics of bovine granulosa cells in vitro. AB - In 24-h cultures, steroid production by cells from non-atretic follicles increased with increasing follicular diameter. Cells from atretic follicles, of all sizes, produced low amounts of oestradiol-17 beta, but very high amounts of progesterone, relative to cells from non-atretic follicles. Increasing the culture period to 72 h caused little change in daily progesterone and oestradiol 17 beta production by granulosa cells from atretic follicles. In contrast, in cells from non-atretic follicles, daily progesterone production increased and daily oestradiol-17 beta production decreased to the levels observed with cells from atretic follicles. Dibutyryl cyclic AMP (1.0 mM) significantly stimulated progesterone production by cells from atretic, but not from non-atretic, follicles. Testosterone (1 microgram/ml) had no effect on progesterone production by cells from atretic follicles, while oestradiol-17 beta, oestrone, testosterone, androstenedione and 5 alpha-dihydro-testosterone (0-1000 ng/ml) each significantly suppressed progesterone production by cells from non-atretic follicles in a dose-dependent manner. Morphometric analysis revealed few subcellular differences between cells from non-atretic and atretic follicles. Mean cell volume was significantly higher for cells from atretic compared to non atretic follicles, but the mean volumes of the major subcellular components were not influenced by follicle health. The mean surface area of the plasma and nuclear membrane, and granular endoplasmic reticulum was also significantly higher in cells from atretic compared to non-atretic follicles. PMID- 3029369 TI - Ovarian follicular activity in Booroola lambs with and without a fecundity gene. AB - The ovaries of 3-month-old Booroola lambs which were heterozygous carriers of a major gene (F) influencing the ovulation rate in mature ewes (i.e. F + lambs) were compared to those ofsimilarly-aged Booroola lambs which were non-carriers of the F-gene (i.e. ++ lambs). The ovaries of the F + Booroola lambs were significantly lighter (P less than 0.01) than those of ++ lambs even though the mean +/- s.e.m. number of follicles (greater than or equal to 1 mm diam.) in the F + lambs was greater than that in the ++ lambs (i.e. F + lambs, 30.2 +/- 2.5 follicles; ++ lambs, 18.4 +/- 1.2 follicles; P less than 0.01). In granulosa cells from non-atretic follicles (greater than or equal to 1 mm diam.) from F + and ++ Booroola lambs, FSH (NIAMDD-FSH-S16) doses of 100 and 1000 ng/ml caused significant stepwise increases (P less than 0.05) in cyclic adenosine 3',5' monophosphate (cAMP) production compared to that achieved at FSH doses of 0 and 1 ng/ml or at any FSH dose in cells from atretic follicles. However, no significant differences in FSH-induced cAMP production were noted with regard to Booroola genotype or follicular diameter. None of the granulosa cell preparations from non atretic follicles of 1-2.5 mm diameter from F + lambs (N = 13) or from non atretic follicles of 1-4.5 mm diameter from ++ lambs (N = 16) responded to LH (NIAMDD-LH-S24; 10 or 1000 ng/ml) to produce significantly more cAMP than did the controls. In contrast, the granulosa cell preparations from non-atretic follicles of 3-4.5 mm diameter from F + lambs (N = 4) and from non-atretic follicles of greater than or equal to 5 mm diameter of ++ lambs (N = 4) produced significantly more cAMP (P less than 0.05) in response to LH (1000 and/or 10 ng/ml) relative to that in the controls. The theca interna from follicles of lambs of both genotypes had functional LH receptors as judged by the androstenedione responses to exogenous LH although no genotypic differences were noted. In F + lambs, the follicular fluid concentrations of testosterone but not oestradiol (i.e. in 1-4.5 mm diam. follicles) and granulosa cell aromatase activity (i.e. in 3-3.5 mm diam. follicles) were significantly higher (both P less than 0.05) than in corresponding follicles or cells from ++ lambs. Collectively the results suggest that the Booroola F-gene influences the composition and function of sheep ovaries before puberty. PMID- 3029370 TI - Calcium pyrophosphate crystals in intermittent hydrarthrosis. AB - A 49-year-old female presented with intermittent hydrarthrosis affecting the right knee. Numerous calcium pyrophosphate crystals were demonstrated in synovial fluid aspirated from the knee. PMID- 3029371 TI - A new thienylpyrazoloquinoline: a potent and orally active inverse agonist to benzodiazepine receptors. PMID- 3029373 TI - The comparative virogenesis of three serotypes of bluetongue virus in Culicoides variipennis (Diptera: Ceratopogonidae). PMID- 3029372 TI - Synthesis of 7,12-dihydropyrido[3,4-b:5,4-b']diindoles. A novel class of rigid, planar benzodiazepine receptor ligands. PMID- 3029374 TI - The effects of smoking and zinc on the oxidative reactions of human neutrophils. AB - Human neutrophils in whole blood and after isolation were examined for their capacity to produce superoxide anions when stimulated by zymosan. The isolation procedure caused neutrophils to lose some of this capacity. The neutrophils of smokers produced less particle-stimulated superoxide than those of non-smokers when the whole blood method was used but not when using isolated cells. Zinc inhibited this aspect of the respiratory burst of isolated neutrophils but enhanced it when the whole blood method was used. We emphasize the importance of methodology when interpreting results. PMID- 3029375 TI - Elevated serum immune complex levels in Pogosta disease, an acute alphavirus infection with rash and arthritis. AB - Circulating immune complexes (CIC) were studied in Pogosta disease, an acute alphavirus infection with fever, rash and arthritis. The disease is caused by a virus antigenically closely related to Sindbis virus. 75 serum specimens from 25 patients with serologically verified infection were obtained from 1-87 days after the onset. Six different CIC detection methods were used and CICs were observed in all patients at least with one test. Tests based on CIC binding onto human platelets followed the natural course of the disease and maximal values were observed between 10-15 days after onset. Slightly elevated levels were observed 2 3 months after onset. The mean conglutinin binding test values were slightly elevated during the whole follow-up period. The severity of arthritis did not directly correlate to CIC levels. C3c and C1q-binding test were positive only in a few cases. Latex and enzyme immunoassay tests for rheumatoid factors gave low positive values in some of the sera. Agarose gel electrophoresis of serum proteins revealed non-specific changes in alpha 1-alpha 2 interzone characteristic of an acute infectious disease. The presence of CIC in the sera of patients with Pogosta disease may indicate body's natural clearange mechanisms of viral antigens. CIC may have a pathogenic role in the prolonged arthritis, even though no direct correlation with CIC levels and severity of arthritis was observed. PMID- 3029376 TI - Complement activation and spleen function in erythema multiforme associated with herpes simplex virus reactivation. AB - Plasma C3d concentrations were measured in patients with erythema multiforme (EM) complicating herpes labialis. C3d levels were raised during cold sores but not during EM. Heat damaged red cell clearance was prolonged during EM. These observations support the concept that immune complexes disseminate herpes simplex virus (HSV) antigens to skin before the onset of EM. PMID- 3029377 TI - Mechanism for leukotriene C4 stimulation of chloride transport in cornea. AB - The leukotriene, LTC4, exerts a stimulatory effect on chloride transport in the frog cornea. In the work described here, the mechanism of action of LTC4 to stimulate chloride transport was studied. In corneas pretreated with indomethacin, the effect of LTC4 was abolished, suggesting the involvement of cyclo-oxygenase products in the response. Incubation of corneas with LTC4 resulted in a significant stimulation in PGE2 synthesis, as determined by TLC autoradiography and radioimmunoassay. In addition, LTC4 was found to stimulate cAMP synthesis in the cornea, and this stimulation was blocked with indomethacin. PGE2 was previously shown by us to be the dominant cyclo-oxygenase product formed in the frog cornea, and is capable of stimulating cAMP and chloride transport. We suggest that LTC4 stimulation of chloride transport is mediated via activation of the cyclo-oxygenase pathway, resulting in enhanced PGE2 synthesis. Elevated PGE2 levels induce cAMP synthesis, and ultimately, the stimulation of chloride transport. Further, the activation of cyclo-oxygenase was found to be dependent on phospholipase A2 activity. This was shown by the inhibition of the LTC4 effect in the presence of quinacrine. Similarly, inhibition of the LTC4 effect in the presence of trifluoperazine suggests that cyclo-oxygenase activation by LTC4 may be mediated via calmodulin. We have previously demonstrated that the frog cornea has the biosynthetic capacity to produce LTC4. Therefore LTC4 may function as an endogenous regulator of chloride transport in this tissue. PMID- 3029378 TI - Proton countertransport by the reconstituted erythrocyte Ca2+-translocating ATPase: evidence using ionophoretic compounds. AB - Human erythrocyte Ca2+-translocating ATPase was solubilized from calmodulin depleted membranes using the detergent Triton X-100, and subsequently purified by calmodulin-affinity chromatography. The purified enzyme was reconstituted in artificial phospholipid vesicles using a cholate-dialysis method and various phospholipids. The reconstituted enzyme was able to translocate Ca2+ inside the vesicles, both in the absence and in the presence of the Ca2+-chelating agent, oxalate, inside the vesicles. The tightness of coupling between ATP hydrolysis and cation translocation was investigated by the use of different ionophoretic compounds. The efficiency of Ca2+ translocation was measured by the ability of the ionophores to stimulate ATP hydrolytic activity of the reconstituted enzyme. It was found that the maximum stimulation of the ATP hydrolytic activity was induced by the electroneutral Ca2+/2H+ ionophore A23187 (9 to 10-fold). A Ca2+ ionophore unable to translocate H+, CYCLEX-2E, was less efficient in stimulating the activity of the reconstituted enzyme (two- to threefold). However, the combined addition of CYCLEX-2E plus protonophores further increased the ATP hydrolytic activity (around fourfold), whereas, the protonophores did not further stimulate ATP hydrolysis in the presence of A23187. Furthermore, in the absence of Ca2+ ionophore, the electroneutral K+(Na+)/H+ ionophoretic exchanger, monensin, stimulated the rate of ATP hydrolysis in the reconstituted enzyme two- or threefold, respectively. These results suggest that the Ca2+-ATPase not only translocates Ca2+ but also H+ in the opposite direction. PMID- 3029379 TI - Complete nucleotide sequence of the topA gene encoding Escherichia coli DNA topoisomerase I. AB - The nucleotide sequence of a 4071 base-pair long segment containing the gene topA encoding Escherichia coli DNA topoisomerase I and its flanking regions has been determined. The gene encodes a total of 864 amino acids from the ATG start to a TAA termination codon, of which the first f-Met appears to be removed after translation; the calculated molecular weight of the translated protein is 97,413. Mapping of promoters by deletion of sequences upstream from the ATG initiation codon indicates the existence of at least two promoters that direct transcription into topA. PMID- 3029380 TI - Probing the structural domains and function in vivo of Escherichia coli DNA topoisomerase I by mutagenesis. AB - Insertion and deletion mutagenesis within the gene topA of Escherichia coli encoding DNA topoisomerase I was carried out to test the existence of subdomains in the enzyme and the relationship between the slow-growth topA- phenotype and the known DNA relaxation activity of the enzyme. All mutants that show no detectable DNA relaxation activity in cell extracts fail to complement the temperature-sensitive growth defect of strain AS17 topAam harboring a plasmid borne temperature-sensitive suppressor tRNA. All mutants that show partial or full levels of DNA relaxation activity in cell extracts (relative to activity in extracts of wild-type cells) can complement this defect. The carboxyl-proximal 25% of the enzyme appears to be in a domain that is dispensable both in terms of the catalytic function of the enzyme and its biological role. Analysis of the mutant enzyme also indicates that the formation of the covalent topoisomerase-DNA complex is correlated with the DNA relaxation activity, which supports the notion that the covalent complex is an obligatory intermediate in the catalysis of DNA topoisomerization. PMID- 3029381 TI - Association of reciprocal exchange with gene conversion between the repeated segments of 2-micron circle. AB - The occurrence of reciprocal exchange of flanking DNA during gene conversion between the repeated segments of the yeast plasmid, 2-micron circle has been examined. The conversion event is induced by making a double-stranded gap within one of the repeats in vitro and allowing the gap to be repaired in vivo. The repair takes place with frequent recombination of flanking markers. Neither the topology of the plasmid substrates (linear or circular) nor the relative orientation of the repeats affects the association rule significantly. These events are reminiscent of meiotic gene conversion between homologous chromosomes but contrast sharply with mitotic or meiotic intrachromosomal gene conversion. It would appear that the difference between the outcomes of intramolecular gene conversion on a chromosome and on a plasmid gapped in vitro does not result from the different physical states of intracellular versus transformed DNA. A gene conversion event in a 2-micron circle : : Tn5 plasmid mediated by the 2-micron circle recombinase (FLP) in vivo, which is formally analogous to the yeast mating type interconversion, often results in recombination of flanking markers. The reaction can be mimicked, in the absence of FLP, by gapping the plasmid within one of the 2-micron circle repeats in vitro and carrying out gap repair in vivo. PMID- 3029382 TI - Functional promoters created by the insertion of transposable element IS1. AB - We have isolated several insertions of the transposable element IS1 into the proximal promoter (P3) of the beta-lactamase gene of plasmid pBR322, which do not abolish resistance to ampicillin. Using a transcription termination module (omega), we have shown that the gene can be expressed from hybrid promoters, created by the insertion of IS1. The terminal inverted repeats of IS1 carry sequences partially homologous to the "-35" consensus region. Splicing either of these sequences to the existing "-10" region of the beta-lactamase promoter by transposition of IS1 at the proper distance results in the formation of an active hybrid promoter. This interpretation was confirmed by transcription studies in vitro. Gene expression from the hybrid promoters was found to be less efficient than from P3. However, the orientation of IS1 that contributes a "-35" with the greater homology to the known "-35" consensus sequence is significantly more efficient than the other. In addition, we were able to assign a strong determinant of IS1 polarity to a 254 base-pair internal segment of IS1. An examination of the ends of many insertion sequences leads us to expect that the phenomenon described here may occur with several of these transposable elements, and may have an unexpected evolutionary significance. PMID- 3029383 TI - Co-ordinate regulation of herpes simplex virus gene expression is mediated by the functional interaction of two immediate early gene products. AB - At early times after infection with herpes simplex virus, transcription from beta promoters is initiated only in the presence of a functional 174,000 Mr phosphoprotein (ICP4), encoded by an immediate early (alpha) gene (IE4). A transient expression assay was used to analyze the requirement for two (ICP4 and ICP0) of the five alpha-gene products in the transcriptional regulation of model alpha and beta-gene promoters. These studies reveal that cells cotransfected with plasmids containing the alpha-gene sequences for infected cell proteins (ICPs) 4 and 0 and a thymidine kinase (TK, a beta-gene) gene or the thymidine kinase promoter fused to a chloramphenicol acetyltransferase (CAT) cassette accumulate 10 to 20-fold more RNA or exhibit 10 to 20-fold more CAT activity than cells cotransfected with a plasmid encoding either alpha-gene protein and a thymidine kinase indicator gene. Functional ICP4 is required for enhanced transcriptional activation in the transient expression assay system. It is also required for the uniform dispersal of ICP0 throughout the nucleus as shown by immunofluorescence staining analysis of transfected cells. Two alpha-promoter-CAT fusions were used as targets to study what effects ICP4, ICP0 and Vmw65 (the virion-associated alpha-gene transactivator) have on expression from alpha-promoters that contain all of the sequences that confer alpha-gene regulation, or only the core sequence governing basal level expression. We conclude that ICP4 can activate alpha-gene expression from the core sequence and, depending on its abundance, activate or repress expression from a promoter containing the sequences required for alpha gene regulation. Independent of these alpha-regulatory sequences cotransfection with low levels of sequences encoding both ICP0 and ICP4 activate expression. At higher ratios of effector (both ICP4 and ICP0) the target accumulation of CAT activity decreases. Although a ts allele of IE4 (cloned from the mutant virus tsK) does not activate alpha-gene expression it can enhance the ability of ICP0 to activate a target containing alpha-regulatory sequences. Virus studies involving tsK support the conclusion that functional ICP4 is required to activate beta-promoters and to repress expression from alpha-promoters and help to explain the pleiotropic effects of the tsK mutation. These analyses have also revealed the presence of a novel RNA species that overlaps the sequences encoding ICP0. Our results suggest that co-ordinate regulation of HSV gene expression is mediated by the functional interaction of at least two alpha-gene products, ICP0 and ICP4. PMID- 3029386 TI - Application of molecular dynamics with interproton distance restraints to three dimensional protein structure determination. A model study of crambin. AB - The applicability of restrained molecular dynamics for the determination of three dimensional protein structures on the basis of short interproton distances (less than 4 A) that can be realistically determined from nuclear magnetic resonance measurements in solution is assessed. The model system used is the 1.2 A resolution crystal structure of the 46 residue protein crambin, from which a set of 240 approximate distance restraints, divided into three ranges (2.5 +/- 0.5, 3.0+0.5(-1.0) and 4 +/- 1 A), is derived. This interproton distance set comprises 159 short-range ([i-j] less than or equal to 5) and 56 ([i-j] greater than 5) long-range inter-residue distances and 25 intra-residue distances. Restrained molecular dynamics are carried out using a number of different protocols starting from two initial structures: a completely extended beta-strand; and an extended structure with two alpha-helices in the same positions as in the crystal structure (residues 7 to 19, and 23 to 30) and all other residues in the form of extended beta-strands. The root-mean-square (r.m.s.) atomic differences between these two initial structures and the crystal structure are 43 A and 23 A, respectively. It is shown that, provided protocols are used that permit the secondary structure elements to form at least partially prior to folding into a tertiary structure, convergence to the correct final structure, both globally and locally, is achieved. The r.m.s. atomic differences between the converged restrained dynamics structures and the crystal structure range from 1.5 to 2.2 A for the backbone atoms and from 2.0 to 2.8 A for all atoms. The r.m.s. atomic difference between the X-ray structure and the structure obtained by first averaging the co-ordinates of the converged restrained dynamics structures is even smaller: 1.0 A for the backbone atoms and 1.6 A for all atoms. These results provide a measure with which to judge future experimental results on proteins whose crystal structures are unknown. In addition, from an examination of the dynamics trajectories, it is shown that the convergence pathways followed by the various simulations are different. PMID- 3029385 TI - Structure of in-vivo transcribing chromatin as studied in simian virus 40 minichromosomes. AB - In order to study the structure of chromatin during transcription, individual in vivo transcribing simian virus 40 (SV40) minichromosomes were analyzed in the electron microscope after crosslinking the nascent RNA strands with different psoralen derivatives to the template DNA. Since psoralen crosslinks the DNA between nucleosomes, spreading of the crosslinked DNA and DNA-RNA complexes reveals single-stranded bubbles at positions where nucleosomes were located. We found that the transcribing SV40 minichromosomes contained a similar number of nucleosomes as did the minichromosomes without crosslinked nascent RNA. The nascent RNA was crosslinked in about equal proportions either in single-stranded bubbles of nucleosomal length or in continuously crosslinked regions between bubbles, in contrast with control experiments with ribosomal chromatin of Dictyostelium. Treatment of SV40 minichromosomes with 1.2 M-NaCl before and during photocrosslinking with psoralen led to the disappearance of the single stranded bubbles. Since no bubbles could be detected at the attachment sites of the RNA molecules when the nucleosomes were disrupted in high salt, and since in about half of the molecules the RNA was attached to fully crosslinked linker DNA, we assume that the single-stranded bubbles with crosslinked RNA are not due to protection by the elongating RNA polymerase II complex, but are rather due to nucleosome-like structures. At the resolution level of single nucleosomes, these results imply for the first time that nucleosome-like structures (perhaps modified compared with "normal" nucleosomes) on SV40 minichromosomes do not prevent transcription elongation by RNA polymerase II. PMID- 3029384 TI - Study of actin filament ends in the human red cell membrane. AB - There is conflicting evidence concerning the state of the actin protofilaments in the membrane cytoskeleton of the human red cell. To resolve this uncertainty, we have analysed their characteristics with respect to nucleation of G-actin polymerization. The effects of cytochalasin E on the rate of elongation of the protofilaments have been measured in a medium containing 0.1 M-sodium chloride and 5 mM-magnesium chloride, using pyrene-labelled G-actin. At an initial monomer concentration far above the critical concentration for the negative ("pointed") end of F-actin, high concentrations of cytochalasin reduce the elongation rate of free F-actin by about 70%. The residual rate is presumed to correspond to the elongation rate at the negative ends. By contrast, the elongation rate on red cell ghosts or cytoskeletons falls to zero, allowing for the background of self nucleated polymerization of the G-actin. The critical concentration of the actin in the red cell membrane has been measured after elongation of the filaments by added pyrenyl-G-actin in the same solvent. It was found to be 0.07 microM, compared with 0.11 microM under the same conditions for actin alone. This is consistent with prediction for the case of blocked negative ends on the red cell actin. The rate of elongation of actin filaments, free and in the red cell membrane cytoskeleton, has been measured as a function of the concentration of an added actin-capping protein, plasma gelsolin, with a high affinity for the positive ends. The elongation rate falls linearly with increasing gelsolin concentration until it approaches a minimum when the gelsolin has bound to all positive filament ends. The elongation rate at this point corresponds to the activity of the negative ends, and its ratio to the unperturbed polymerization rate (in the absence of capping proteins) is indistinguishable from zero in the case of ghosts, but about 1 : 4 in the case of F-actin. When ATP is replaced in the system by ADP, so that the critical concentrations at the two filament ends are equalized, the difference is equally well-marked: for F-actin, the rate at the equivalence point is about 40% of that in the absence of capping protein, whereas for ghosts the nucleated polymerization rate at the equivalence point is again zero, indicating that under these conditions the negative ends contribute little or not at all to the rate of elongation.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3029387 TI - Solution structure of human growth hormone releasing factor. Combined use of circular dichroism and nuclear magnetic resonance spectroscopy. AB - The solution structures of two human growth hormone releasing factor analogues, 27Leu45Gly-hGHRF(1-45)OH and 27Nle-hGHRF(1-29)NH2, are investigated by means of circular dichroism and nuclear magnetic resonance spectroscopy. Using circular dichroism spectroscopy, it is shown that both peptides adopt ordered structures at low concentrations of trifluoroethanol (approximately 30%). Quantitative analysis of the circular dichroism spectra indicates that the same number of residues, approximately 23 to 25, are in a helical state in both peptides. Using two-dimensional nuclear magnetic resonance methods all proton resonances of the 27Nle-hGHRF(1-29)NH2 fragment are assigned and its secondary structure is determined from a qualitative interpretation of the nuclear Overhauser enhancement data. Two distinctive regions of alpha-helix are present extending from residues 6 to 13 and 16 to 29. PMID- 3029388 TI - Molecular analysis of insertional mutations in the yellow gene region of Drosophila. AB - We have cloned 70 kb of DNA from the yellow (y) gene region and analyzed two y null alleles. These alleles are caused by different DNA elements that have inserted into different sites of the y gene coding region. PMID- 3029389 TI - Crystallization and preliminary X-ray diffraction study of ferrocytochrome c2 from Rhodopseudomonas viridis. AB - Crystals of ferrocytochrome c2 from a non-sulphur purple photosynthetic bacterium, Rhodopseudomonas viridis, have been grown from ammonium sulphate solution at pH 8.5 by the sitting-drop vapour-diffusion procedure. The crystals belong to the trigonal system, space group P3(1)21 (or its enantiomorph P3(2)21) with unit-cell dimensions of a = b = 75.8 A and c = 40.1 A, and diffract to at least 2.0 A resolution. Assuming that an asymmetric unit contains one protein molecule (approx. 12,300 Mr), the solvent content of the crystal is approximately 54.5% (v/v). PMID- 3029390 TI - Structure and function of the F plasmid genes essential for partitioning. AB - The F plasmid in Escherichia coli has its own partition mechanism controlled by the sopA and sopB genes, and by the cis-acting sopC region. The DNA sequence of the entire partition region and its flanking regions is described here. Two large open reading frames coding for 43,700 Mr and 35,400 Mr proteins correspond to sopA and sopB, respectively. The sopB reading frame is located immediately downstream from the sopA reading frame. Twelve 43 base-pair direct repeats exist in the sopC region without any spacer regions, and one pair of seven base-pair inverted repeats exists in each of the direct repeats. Analysis of deletions in the sopC region showed that the direct repeats play an important role in plasmid partition and IncD incompatibility. IncG incompatibility is exhibited by pBR322 derivatives carrying the sopB gene alone. When compared with the partition genes parA and parB of plasmid P1, homology in amino acid sequence was found between the SopA protein of F and the ParA protein of P1, and also between SopB protein of F and ParB protein of P1. In addition, homology was found between Rep proteins of F and P1. PMID- 3029391 TI - Sarcolemmal Na-Ca exchange and sarcoplasmic reticulum calcium uptake in developing chick heart. AB - Ontogenetic changes in calcium transport mediated by the sarcolemmal Na-Ca exchanger and by the sarcoplasmic reticulum calcium pump were studied in crude membranes from chick heart. Transport activities were evaluated per mass of membrane protein and heart tissue. Relative to unit heart mass Na-Ca exchange activity increases linearly from embryonic day 4 to day 10 of newborn stage. The overall increase is about 20-fold. An excellent correlation exists between activity of sodium gradient-induced calcium uptake and ouabain-sensitive (Na,K) ATPase in crude membranes of embryonic, newborn and adult hearts. In the same membrane preparations active calcium uptake into vesicles of sarcoplasmic reticulum increases about 3-fold from embryonic day 4 to embryonic day 7, and then increases continuously until day 20. This is followed by a 3-fold elevation in reticular calcium accumulation at hatching on day 21. Maximal sarcoplasmic reticulum calcium transport activity reached at day 10 after hatching is 40- to 50-fold greater than activity values at embryonic day 4. In adult hearts the activities of both Na-Ca exchange and reticular calcium uptake drop to levels characteristic for the late embryonic period. This comparative study of sarcolemmal sodium gradient-dependent calcium flux and reticular calcium sequestration demonstrates that during chick heart differentiation the two calcium transport systems do not develop in parallel. Na-Ca exchange appears to play a greater role in calcium control of embryonic as compared to newborn and adult hearts. By contrast, the contribution of sarcoplasmic reticulum calcium transport to cardiac calcium movements becomes more predominant during and after hatching. PMID- 3029392 TI - ACTH-producing apudoma metastatic to the liver. AB - A young man presented with combative psychosis and elevated levels of plasma adrenocorticotropic hormone (ACTH). A solitary vascular pancreatic mass and diffuse vascular hepatic nodules were demonstrated on selective splenic and hepatic arteriograms. These classic angiographic findings are used to emphasize the role of angiography in initial radiographic evaluation and to summarize the angiographic appearance of functioning pancreatic adenomas. Even though this is an aggressive tumor, early diagnosis and intensive treatment may allow prolonged remission, if not cure. PMID- 3029393 TI - Urinary excretion of 2,5-hexanedione and peripheral polyneuropathies workers exposed to hexane. AB - Forty shoe factory workers who were exposed to hexane were investigated to see if there was a correlation between electroneuromyographic changes indicative of neuropathy and urinary excretion of 2,5-hexanedione. Urinary samples were analyzed for the presence of the metabolic products of n-hexane and its isomers. Electrodiagnostic examination was carried out following the urinary sampling. A rating scale was used to obtain a cumulative numeric index of electrodiagnostic findings. 2,5-Hexanedione and gamma-valerolactone were discovered in all cases, while 2-hexanol was found in 11 cases. 2,5-Hexanedione was the main metabolite in most cases (39 of 40). Only in 1 case was a low level of 2-methyl-2-pentanol detected; 3-methyl-2-pentanol was never detected. Metabolic products of cyclohexane were present in about one-fifth of the cases, while trichloroethanol, a metabolic product of trichoroethylene, was nearly always present, all at very low concentrations. Electromyographic abnormalities significant for early detection of toxic polyneuropathy were found in 14 cases. A statistically significant correlation of the electroneuromyographic scoring on the urinary concentrations of measured metabolites was observed only with 2,5-hexanedione and gamma-valerolactone, both derived from n-hexane. Since gamma-valerolactone is probably not a true metabolite of n-hexane, our results support the hypothesis that polyneuropathies in shoemakers are due to 2,5-hexanedione. For practical purposes the urinary concentration of 2,5-hexanedione can serve as a predictive measurement for early detection of neurotoxic lesions at preclinical states. PMID- 3029394 TI - Oxygen-derived free radical inhibition in the healing of experimental zone-of stasis burns. AB - When dehydration, infection, and mechanical trauma are prevented, procedures (such as cooling and/or oral antithromboxane) designed to diminish ischemia in experimental zone-of-stasis burns have been associated with no or only minor improvement in wound healing. To test the hypothesis that ongoing skin damage occurring postburn (PB) may in part be due to release of oxygen-derived free radicals during the 16-hour through 4-day PB period of reperfusion in such burns, beginning immediately and for a period of 5 days PB, equal numbers of guinea pigs received: allopurinol 150 mg/kg PO q 6 h vs. placebo, dimethylsulfoxide (DMSO) 75% applied topically q 12 h vs. placebo, or yeast-derived superoxide dismutase coupled with polyethylene glycol (PEG-SOD, Pharmacia) 10,000 U (Fridovich) given IV q 8 h producing a concentration of 16 U/cc of plasma 8 hr after injection vs. placebo. Gross and histologic examination of wounds by a 'blinded' investigator at 1 week and 3 weeks PB revealed no difference between treatment and control groups when rates of re-epithelialization and frequencies of hair-follicle retention were compared. Using the dosages, routes, and model described, treatment of a zone-of-stasis burn with PO allopurinol (a xanthine oxidase inhibitor), topical DMSO (a scavenger of the hydroxyl radical), or IV PEG-SOD (a scavenger of the superoxide radical) during the first 5 days PB was associated with no increase in the rate of re-epithelialization or frequency of hair follicle retention at 1 and 3 weeks PB when compared with controls. PMID- 3029395 TI - Serological survey for human monkeypox infections in a selected population in Zaire. AB - About 3460 persons living in Kole zone of East Kasai, in Zaire, were examined and their sera screened initially by a haemagglutination-inhibition test. Of these, 667 (19%) were positive. Radioimmunoassay adsorption tests for the presence of monkeypox- or vaccinia-specific antibodies gave unequivocal results in 300 of these sera; the remaining 47 were nonspecific. Monkeypox-specific antibodies were found in sera of 27 individuals, of all ages and both sexes, giving an overall prevalence rate of monkeypox virus-specific antibodies of 0.8%. The prevalence rate was four times higher in the 5 to 9 year age group (1.3%) than in children aged 0 to 4 years (0.3%), and was highest (2.4%) in the 15 to 19 year age group. There was no significant difference in the prevalence rates between the sexes. As might be expected, there are substantially higher prevalence rates in persons living in forest galleries than in those in savannah, and among those living in areas where human monkeypox cases had occurred in the past compared with those living in other localities. Nineteen children whose sera showed specific monkeypox antibodies were re-examined. Twelve showed facial and body skin changes suggesting the presence of vesiculo-pustular disease in the past; four of these had been known registered monkeypox cases. Seven children had neither signs nor history of past vesiculo-pustular disease, suggesting that they had suffered from subclinical infection with monkeypox virus. PMID- 3029397 TI - Molecular epizootiology and evolution of vesicular stomatitis virus New Jersey. AB - Vesicular stomatitis virus (VSV) has been shown previously to be capable of undergoing rapid mutational change during sequential experimental infections in various tissue culture cell systems (J. Holland, K. Spindler, F. Horodyski, E. Grabau, S. Nichol, and S. Vandepol, Science 215:1577-1585, 1982). The present study was undertaken to determine the degree of genetic diversity and evolution of the virus under natural infection conditions and to gain insight into the epizootiology of the disease. Between 1982 and 1985, numerous outbreaks of VSV of the New Jersey serotype were reported throughout regions of the United States and Mexico. A T1 RNase fingerprint analysis was performed on the RNA genomes of 43 virus isolates from areas of epizootic and enzootic virus activity. This indicates that virus populations were genetically relatively homogeneous within successive U.S. virus epizootics. The data included virus isolates from different epizootic stages, geographical locations, host animals, and host lesion sites. In contrast, only distant genome RNA T1 fingerprint similarities were observed among viruses of the different U.S. epizootics. However, Mexican viruses isolated before or concurrent with U.S. epizootics had very similar RNA genome fingerprints, suggesting that Mexico may have been the possible origin of virus initiating recent U.S. VSV New Jersey outbreaks. Comparison of T1 fingerprints of viruses with enzootic disease areas revealed a greater extent of virus genetic diversity in these areas relative to that observed in epizootic areas. The evolutionary significance of these findings and their relationship to experimental data on VSV evolution are discussed. PMID- 3029396 TI - Modification of RNA polymerase IIO subspecies after poliovirus infection. AB - Infection of HeLa cells with poliovirus results in a shutdown of host transcription. In an effort to understand the mechanism(s) that underlies this process, we analyzed the distribution of RNA polymerase IIO before and after viral infection. Analysis of free and chromatin-bound enzyme indicated that there is a significant reduction in RNA polymerase IIO following infection. This observation, together with increasing evidence that transcription is catalyzed by RNA polymerase IIO, supports the hypothesis that poliovirus-induced inhibition of host transcription occurs at the level of RNA chain initiation and involves the direct modification of RNA polymerase II. PMID- 3029398 TI - Horizontal transmission of murine retroviruses. AB - Both a feral mouse ecotropic virus (WM-E) and Friend ecotropic virus (F-MuLV) were transmitted horizontally among adult mice. Infection resulted in the production of antiviral antibody in the recipients, with no evidence of viremia or clinical disease. However, persistent low-level virus replication was detectable in the spleens of these mice as long as 8 months after initial infection. External secretions, including saliva, semen, and uterine secretions from viremic mice contained high concentrations of infectious virus. Nevertheless, transmission occurred only from viremic males to either males or females. Male-to-male transmission appeared to occur by parenteral inoculation of infectious saliva during fighting behavior. Evidence is presented that infection of females was by the venereal route. Of four mouse strains examined, NFS/N, IRW, and C57L females were all susceptible to venereal infection, whereas AKR mice were not. Since AKR mice are susceptible to infection by WM-E administered parenterally, this resistance appeared to be mediated by local viral interference due to the high-level expression of endogenous Akv gp70 within the female reproductive tract. Although both WM-E and F-MuLV were transmitted from viremic males to females, infection by WM-E was significantly more efficient than that by F-MuLV. This difference correlated with a distinct difference in cellular tropism of WM-E and F-MuLV within the epididymis of viremic males. F-MuLV gp70 was expressed only within stromal elements, whereas WM-E gp70 was seen largely within the epithelial lining cells and luminal contents of the duct. No evidence of virus expression within germ cells was observed. The possible influence of virus expression by epithelial cells of the female reproductive tract on infection of embryos is discussed. PMID- 3029399 TI - Vaccinia virus recombinants expressing the SA11 rotavirus VP7 glycoprotein gene induce serotype-specific neutralizing antibodies. AB - A cDNA copy of the gene coding for the major outer neutralizing protein (VP7) of simian 11 rotavirus was incorporated into the vaccinia virus genome under the control of the vaccinia promoter (molecular weight, 7,500). A deletion mutant of this gene which codes for a secreted form of VP7 when expressed under the control of the simian virus 40 late promoter (M. S. Poruschynsky, C. Tyndall, G. W. Both, F. Sato, A. R. Bellamy, and P. H. Atkinson, J. Cell Biol. 101:2199-2209, 1985) was also inserted. Each recombinant vaccinia virus directed the synthesis of a rotavirus protein in infected cells, and the product encoded by the mutated gene was secreted. Rabbits immunized with the two types of recombinant vaccinia virus generated antibodies that were able both to recognize simian 11 rotavirus in an enzyme-linked immunosorbent assay and to neutralize the virus in a plaque reduction test. Antibodies induced by the recombinant vaccinia viruses expressing either form of VP7 were serotype specific. PMID- 3029400 TI - Correlation of leukemogenic potential of murine retroviruses with transcriptional tissue preference of the viral long terminal repeats. AB - Recombination studies have established that retroviral long terminal repeats (LTRs) are important genetic determinants of the viral capacity to induce hematopoietic tumors and to specify the type of cell making up the tumor. Plasmids containing LTRs of several murine leukemia viruses linked to the chloramphenicol acetyltransferase gene were tested in transient assays to measure relative rates of transcriptional activity in different types of hematopoietic cells. LTRs of the thymomagenic viruses SL3-3, Moloney leukemia virus, and a Moloney mink cell focus-forming virus all expressed to higher levels than other LTRs in T-lymphocyte cell lines. Conversely, the LTRs of Friend leukemia virus and a polycythemic spleen focus-forming virus expressed to higher levels than other LTRs in erythroleukemia cells. The LTR of nonleukemogenic Akv virus induced a relatively low level of activity compared with the others in all cells tested. Thus the relative level of LTR-driven expression in various types of cells corresponds to the type of tumor caused by the intact virus in vivo. These results provide direct evidence that the tissue specificity of the transcriptional activity of LTRs plays a critical role in determining the target cell for retroviral oncogenesis. PMID- 3029401 TI - Activation of int-1 and int-2 mammary oncogenes in hormone-dependent and independent mammary tumors of GR mice. AB - Mammary tumors in the GR mouse strain are caused by the expression of an endogenous provirus of the mouse mammary tumor virus (MMTV). The tumors progress from a hormone-dependent growth phase to autonomous, hormone-independent growth. We studied proviral insertion of MMTV at the int-1 and int-2 mammary oncogenes and the transcription of these genes during progression of a series of transplanted mammary tumors. During the hormone-dependent phase, 6 of 15 transplanted tumors were positive for proviral insertion at int-1 or int-2 or both. These tumors were oligoclonal with respect to the fraction of tumor cells with novel int-1 and int-2 restriction fragments and, apparently, consisted of different tumor cells with proviruses integrated at different oncogenes, including genes that are not yet known. In 10 tumors we detected expression of the int genes, indicating that most tumors contain minor populations of cells with int-1 or int-2 activations. On transplantation the tumors remained oligoclonal during the hormone-dependent phase. The hormone-independent variants of the tumors emerged as clonal outgrowths of cells with MMTV proviruses that could be traced back in the hormone-dependent tumors, but not always those of cells that were positive for insertions near int-1 or int-2. The maintenance of oligoclonality during the hormone-dependent phase suggests a growth-controlling effect of different populations of cells on each other. The clonal, hormone independent tumors that arise later seem to be the result of mutations that are unrelated to int activation. PMID- 3029402 TI - Analysis of mixed infection of sheep with bluetongue virus serotypes 10 and 17: evidence for genetic reassortment in the vertebrate host. AB - Two seronegative sheep were infected intravenously with 10(9) PFU each of bluetongue virus (BTV) serotype 10 and BTV serotype 17. One animal experienced a mild bluetongue-like disease, and both experienced a short-duration viremia and developed neutralizing immune responses to both virus serotypes. Progeny virus was isolated from venous blood from each animal by using conditions in which reassortment could not have occurred during isolation. Electropherotypes were determined for the progeny viruses from the infected sheep, yielding strikingly similar results for the two animals. In both sheep, serotype 10 dominated among the progeny, accounting for 92% of the progeny. Serotype 17 was rarely isolated and accounted for 3% of the progeny analyzed. The remaining 5% of the progeny clones were reassortant and derived genome segments from both serotypes 10 and 17. Analysis of the parental origin of genome segments in the small number of reassortant progeny analyzed suggested that selection of specific genome segments may have occurred in the infected sheep. These data indicate that reassortment of genome segments occurs, at low frequency, in sheep mixedly infected with BTV. PMID- 3029403 TI - Mutants defective in herpes simplex virus type 2 ICP4: isolation and preliminary characterization. AB - Vero cells were biochemically transformed with the gene encoding ICP4 of herpes simplex virus type 2 (HSV-2). These cells were used as permissive hosts to isolate and propagate HSV-2 mutants defective in this gene. Two mutants, designated hr259 and hr79, were isolated. Neither mutant grew in nontransformed Vero cells, but both grew to near wild-type levels in HSV-2 ICP4-expressing cells. hr259 contains a deletion of about 0.6 kilobases which eliminates the mRNA start site of the ICP4 gene. hr79 contains a mutation which maps by marker rescue to the portion of the ICP4 gene encoding the carboxy-terminal half of the protein. Although hr259 failed to generate any detectable ICP4 mRNA in nontransformed Vero cells, hr79 encoded an ICP4 mRNA which is wild type with respect to size. In nontransformed Vero cells infected with hr259, only ICP0, ICP6, ICP22, and ICP27 were readily detectable. In the case of hr79, a truncated form of ICP4 appeared to be made in addition to ICP0, ICP6, ICP22, and ICP27. Both hr259 and hr79 grew efficiently on cell lines transformed with the ICP4 gene of HSV-1 as evidenced by plating efficiencies and single-burst experiments. Similarly, cells transformed with the ICP4 gene of HSV-2 served as efficient hosts for the growth of d120, HSV-1 ICP4 deletion mutant. PMID- 3029404 TI - Effects of a temperature-sensitive mutation in the immediate-early gene of pseudorabies virus on class II and class III gene transcription. AB - The pseudorabies virus immediate-early protein activates transcription of both class II and class III genes. At present it is not known whether the activation of class II genes occurs through an activation of cellular factors or by a direct interaction of the immediate-early protein with factors or DNA. It is also not known whether the activation of class II and class III genes occurs by a similar or different mechanism. We utilized tsG, a temperature-sensitive mutation in the immediate-early gene of pseudorabies virus, to study the activation of viral genes transcribed by RNA polymerases II and III. Previous studies have shown that tsG inhibits wild-type adenovirus early gene transcription in coinfections at the nonpermissive temperature (L. T. Feldman and S. E. Ahlers, J. Virol. 57:13-17, 1986). Using this system of mixed infections as an assay, we obtained several results which allowed us to draw certain conclusions about the mode of action of the pseudorabies virus immediate-early protein (IEP). First, the tsG mutation inhibits the formation of new transcription complexes on class II genes, but does not affect transcription from preestablished transcription complexes formed in the presence or absence of the adenovirus E1A protein. Second, tsG does not inhibit transcription from class III genes, suggesting that the activation by IEP of class II and class III genes occurs by different mechanisms. Third, activation of transcription by the adenovirus E1A protein is not dominant to inhibition by tsG, suggesting that the temperature-sensitive IEP is involved in some physical interaction which actively inhibits transcription of viral genes. PMID- 3029405 TI - Murine cells carrying integrated tandem genomes of hepatitis B virus DNA transcribe RNAs from endogenous promoters on both viral strands and express middle and major viral envelope proteins. AB - Clone 4.10 cells were isolated as a methotrexate-resistant clone arising after cotransfection of mouse 3T3 cells with plasmid DNA containing a head-to-tail dimer of the hepatitis B virus (HBV) genome and DNA coding for methotrexate resistant dihydrofolate reductase. The majority of methotrexate-resistant clones derived by this procedure have been found to contain multiple copies of the HBV genome, but the intact HBV dimer was rarely preserved. In contrast, 4.10 cells contained at least 40 copies of intact HBV dimer per cell. These cells produced large amounts of 22-nm hepatitis B surface antigen particles that included viral envelope proteins reactive with the pre-S2 region-specific antibody, indicating transcription and translation of the pre-S2 and S regions of the integrated viral genomes. The cells also synthesized viral e antigen, which was released into the culture medium. Characterization of polyadenylated viral RNAs transcribed from the long (minus) strand of the integrated HBV DNA demonstrated the presence of shorter-than-genome-length RNAs containing only X region sequences, shorter-than genome-length RNAs containing both X and S region sequences, and longer-than genome-length RNAs containing core, X, and S region sequences. Start sites for transcripts were mapped 5' to and within the pre-S region and 5' to and within the precore region at approximately the same sites as those utilized for HBV transcription during viral replication in infected livers. Polyadenylated RNA transcripts complementary to the short (plus) strand of HBV that initiated and terminated within the intact and integrated head-to-tail tandem viral genomes were also detected. PMID- 3029406 TI - Chemical and immunological characterizations of equine infectious anemia virus gag-encoded proteins. AB - The viral core proteins (p15, p26, p11, and p9) of equine infectious anemia virus (EIAV) (Wyoming strain) were purified by reverse-phase high-pressure liquid chromatography. Each purified protein was analyzed for amino acid content, N terminal amino acid sequence, C-terminal amino acid sequence, and phosphoamino acid content. The results of N- and C-terminal amino acid sequence analysis of each gag protein, taken together with the nucleotide sequence of the EIAV gag gene (R. M. Stephens, J. W. Casey, and N. R. Rice, Science 231:589-594, 1986), show that the order of the proteins in the precursor is p15-p26-*-p11-p9, where a pentapeptide also found in the virus is represented by the asterisk. The data are in complete agreement with the predicted structure of the gag polyprotein and show the peptide bonds cleaved during proteolytic processing. The N-terminus of p15 is blocked to Edman degradation. The p11 protein is identical to the nucleic acid-binding protein of EIAV previously isolated (C. W. Long, L. E. Henderson, and S. Oroszlan, Virology 104:491-496, 1980). High-titer rabbit antiserum was prepared against each purified protein. These antisera were used to detect the putative gag precursor (Pr55gag) and intermediate cleavage products designated Pr49 (p15-p26-*-p11), Pr40 (p15-p26), and Pr35 (p26-*-p11) in the virus and in virus-infected cells. High-titer antisera to EIAV p15 and p26 showed cross reactivity with the homologous protein of human T-cell lymphotropic virus type III/lymphadenopathy-associated virus. PMID- 3029407 TI - Antigenic cross-reactions among herpes simplex virus types 1 and 2, Epstein-Barr virus, and cytomegalovirus. AB - Polyvalent rabbit antisera against herpes simplex virus type 1 and 2 (HSV-1 and HSV-2), cytomegalovirus (CMV), and Epstein-Barr virus (EBV), monospecific antisera against affinity-purified HSV-2 glycoproteins gB and gG, and a panel of monoclonal antibodies against HSV and EBV proteins were used to analyze cross reactive molecules in cells infected with the four herpesviruses. A combination of immunoprecipitation and Western blotting with these reagents was used to determine that all four viruses coded for a glycoprotein that cross-reacted with HSV-1 gB. CMV coded for proteins that cross-reacted with HSV-2 gC, gD, and gE. Both CMV and EBV coded for proteins that cross-reacted with HSV-2 gG. Antigenic counterparts to the p45 nucleocapsid protein of HSV-2 were present in HSV-1 and CMV, and counterparts of the major DNA-binding protein and the ribonucleotide reductase of HSV-1 were present in all the viruses. The EBV virion glycoprotein gp85 was immunoprecipitated by antisera to HSV-1, HSV-2, and CMV. Antisera to CMV and EBV neutralized the infectivity of both HSV-1 and HSV-2 at high concentrations. This suggests that cross-reactivity between these four human herpesviruses may have pathogenic as well as evolutionary significance. PMID- 3029408 TI - Expression of herpes simplex virus type 1 major DNA-binding protein, ICP8, in transformed cell lines: complementation of deletion mutants and inhibition of wild-type virus. AB - To minimize the contribution of residual activity associated with the temperature sensitive (ts) form of ICP8 specified by available ts mutants, deletion mutations in this gene were constructed. Cells permissive for the generation and propagation of ICP8 deletion mutants were first obtained. Vero cells were cotransfected with pKEF-P4, which contains the gene for ICP8, and pSV2neo or a hybrid plasmid containing the G418 resistance gene linked to pKEF-P4. Of the 48 G418-resistant cell lines, 21 complemented ICP8 ts mutants in plaque assays at the nonpermissive temperature. Four of these were examined by Southern blot analysis and shown to contain 1 to 3 copies of the ICP8 gene per haploid genome equivalent. Cell line U-47 was used as the permissive host for construction of ICP8 deletion mutants. In addition to cell lines which complemented ts mutants, two lines, U-27 and U-35, significantly inhibited plaque formation by wild-type virus, contained 30 and 100 copies of the ICP8 gene per haploid genome equivalent, respectively, and expressed large amounts of ICP8 after infection with wild-type virus. At low but not high multiplicities of infection, this inhibition was accompanied by underproduction of viral polypeptides of the early, delayed-early, and late kinetic classes. For construction of deletion mutants, a 780-base-pair XhoI fragment was deleted from pSG18-SalIA, a plasmid which contains the gene for ICP8, to yield pDX. U-47 cells were then cotransfected with pDX and infectious wild-type DNA. Mutant d61, isolated from the progeny of cotransfection, was found to contain both the engineered deletion in the ICP8 gene and an oriL-associated deletion of approximately 55 base pairs. Because d61 contained two mutations, a second mutant, d21, which carried the engineered ICP8 deletion but an intact oriL, was constructed by cotransfection of U-47 cells with wild-type DNA and an SalI-KpnI fragment purified from pDX. Phenotypic analysis of d21 and d61 revealed that they were similar in all properties examined: both exhibited efficient growth in U-47 cells but not in Vero cells; both induced the synthesis of an ICP8 polypeptide which was smaller than the wild-type form of the protein and which, unlike the wild-type protein, was found in the cytoplasm and not the nucleus of infected Vero cells; and nonpermissive Vero cells infected with either mutant failed to express late viral polypeptides. PMID- 3029409 TI - Phosphorylation of polyomavirus large T antigen: effects of viral mutations and cell growth state. AB - Phosphorylation is responsible for the shift in electrophoretic mobility of polyomavirus large T antigen observed in pulse-chase or continuous-labeling experiments. Phosphorylated forms migrated more slowly than newly synthesized [35S]methionine large T antigen, and alkaline phosphatase treatment reversed the mobility shift. Analysis of phosphopeptides with Staphylococcus aureus V8 protease showed that large T antigen forms of intermediate mobility were enriched in peptides 1 to 4, 8, and 9, while the slower migrating species had all nine phosphopeptides, including peptides 5 and 7. The phosphorylations represented by phosphopeptides 5 and 7 were of particular interest. These phosphopeptides were entirely lacking in large T antigen from tsa mutants such as ts616 labeled at the nonpermissive temperature. Also, the phosphorylation of peptides 5 and 7 depends on the growth state of the cell. Early in infection of quiescent cells intermediate mobility forms of large T antigen with little or no phosphorylation, particularly of peptides 5 and 7, were seen, whereas peptides 5 and 7 were well represented at the same time in patterns from growing cells. Later in infection of growth-arrested cells, these phosphorylations were observed, suggesting that infection stimulates the relevant kinase. Because large T antigen of hrt mutants, which lack middle and small T antigens, showed phosphorylation of peptides 5 and 7, large T antigen was apparently responsible for the stimulation. Because some differences in the distribution of phosphopeptides were noted between hrt mutants and the wild type, middle T antigen, small T antigen, or both may play a modulating role in large T antigen phosphorylation. PMID- 3029410 TI - Localization of the phosphorylations of polyomavirus large T antigen. AB - Polyomavirus large T antigen is phosphorylated on both serine and threonine residues at a ratio of approximately 6 to 1. This phosphorylation could be resolved into a series of nine Staphylococcus aureus V8 phosphopeptides. All of these were found in an N-terminal chymotryptic fragment with a molecular weight of 57,000. A C-terminal formic acid fragment of 50,000-molecular-weight lacked phosphate. Therefore, unlike simian virus 40 large T antigen, polyomavirus large T antigen has no significant C-terminal phosphorylation. Limited V8 and hydroxylamine cleavage showed that the phosphorylations can be localized to two different portions of the molecule. A significant fraction of the phosphate was localized in the N-terminal portion of the molecule before residue 183. Within this region V8 peptides 4, 8, and 9 represented phosphorylations that were more proximal, while peptides 1, 2, and 3 included more distal phosphorylations. None of these phosphorylations appeared analogous to those of simian virus 40 large T antigen. V8 phosphopeptides 5 and 7 were more distal and could be distinguished in biological experiments from the N-terminal phosphorylations. Formic acid mapping suggested that much, if not all, of this phosphorylation is located between residues 257 and 285. PMID- 3029411 TI - Dsi-1, a region with frequent proviral insertions in Moloney murine leukemia virus-induced rat thymomas. AB - Dsi-1 is a region of chromosomal DNA that underwent proviral insertion in 3 of 24 Moloney murine leukemia virus-induced rat thymomas. In one of these tumors, a provirus is also integrated adjacent to the proto-oncogene c-myc. The proviruses in Dsi-1 have been characterized and appear to be complete. The proviruses were located within a 2-kilobase region that contained four prominent DNase I hypersensitive sites. These hypersensitive sites were observed in Moloney murine leukemia virus-induced thymomas but not in NRK cells. The region of Dsi-1 immediately 3' to the insertions cross-hybridized with human and chicken DNA, indicating that it contains highly conserved sequences. No evidence could be found for the expression of this highly conserved region. Dsi-1 was mapped to mouse chromosome 4. This location demonstrates that Dsi-1 is different from 16 of the known proto-oncogenes (c-abl, c-erbA c-erbB, c-ets-1, c-ets-2, c-fes, c-fos, c-myb, c-myc, c-raf, A-raf, c-Ha-ras, c-Ki-ras, N-ras, c-sis, and c-src) and 12 cellular regions of tumor-associated integrations in retrovirus-induced tumors (c erbB, Fis-1, int-1, int-2, Mis-1/pvt-1, Mlvi-1, Mlvi-2, c-mos, c-myb, c-myc, Pim 1, and c-Ha-ras). Hybridization experiments indicated that Dsi-1 is probably different from five additional proto-oncogenes (c-fgr, c-fms, c-mos, neu, and c yes) and from two additional frequent integration regions (lck and Mlvi-3). PMID- 3029413 TI - Integration of hepatitis B virus: analysis of unoccupied sites. AB - Hepatitis B virus (HBV) sequences integrated in the PLC/PRF/5 cell line (Alexander cells), which was derived from a human primary liver carcinoma, were previously extensively studied. Here we describe the analysis of the unoccupied sites of two linearly integrated forms of HBV DNA, AL-14 and AL-26, that were characterized previously. No major cellular DNA rearrangements were seen at the integration sites except for small deletions of host sequences: 2 kilobases of DNA in AL-14 and 17 base pairs (bp) in AL-26. The unoccupied site of AL-26 was found to be missing 182 bp, which previously mapped next to the right end of the integration sites of several independent clones. These were believed to be of cellular origin, but we show here that these 182 bp are in fact from unusual HBV sequences. Surprisingly, a region of this newly detected HBV DNA sequence is more homologous to that of woodchuck HBV DNA. Our analysis shows that the normal counterparts of both AL-14 and AL-26 contain minisatellite-like repetitive sequences. Based on the data presented here and our previous finding of HBV DNA integration at satellite sequences, we propose that genomic simple repetitive sequences are hot spots for HBV DNA integration. PMID- 3029412 TI - Characterization of Rous sarcoma virus sequences essential for viral gene expression. AB - Using the Escherichia coli lacZ gene product beta-galactosidase as an indicator of gene expression, we analyzed sequences that are required for expression of the Rous sarcoma virus (RSV) genome in avian cells. The RSV long terminal repeat (LTR) and leader region were sufficient to direct the synthesis of high levels of enzymatically active gag-lacZ fusion proteins. A portion of U3 greater than 140 nucleotides upstream from the cap site was essential for gene expression. This element functioned in either orientation, but its activity was attenuated when it was relocated further away from the cap site. The insertion of exogenous LTRs 3' of lacZ augmented the expression of that gene by increasing the level of stable gag-lacZ transcripts. Furthermore, 3' LTRs could partially compensate for certain defects within the 5' LTR. Insertion of various fragmentary LTRs allowed the identification of at least three synergistically acting domains within the 3' LTR that influence gene expression. Interestingly, the gag-lacZ expression was only stimulated by a 3' LTR when the exogenous 3'-untranslated region was adjacent. Our results imply that the two LTRs of a provirus interact in a complex manner to promote high levels of stable transcripts. It was also found that gag-lacZ expression was independent of viral gene products, suggesting that trans activation is not a key mechanism regulating RSV expression in avian cells. PMID- 3029414 TI - Transition from a heterozygous to a homozygous state of a pair of loci in the inverted repeat sequences of the L component of the herpes simplex virus type 1 genome. AB - The behavior of herpes simplex virus type 1 heterozygous isolates, in which the two inverted repeats of the L component (RL) were differentiated by a polymorphism marker (the presence [type B] or absence [type A] of a SalI site), was investigated. The progeny viruses derived from the heterozygote (A/B) consisted of heterozygotes (A/B), type A homozygotes (A/A), and type B homozygotes (B/B). The heterology between RL, albeit tolerated, was unstable, as is the case with heterology between the repeats of the S component. The two repeats TRL (terminal) and IRL (internal) were equipotent in generating homozygotes from a heterozygote. Data obtained from an analysis of 426 progeny viruses derived from heterozygous clones supported the hypothesis that the two loci in RL of a herpes simplex virus type 1 genome are determined as a random combination of the corresponding two loci in RL of the parent virus and that the ratio of heterozygotes/type A homozygotes/type B homozygotes in the progeny viruses from a heterozygote is expected to be 2:1:1. An ephemeral dominance of one type of homozygote over the other was observed in subclones from several heterozygous clones. PMID- 3029415 TI - Nucleic acid sequence and oncogenic properties of the HZ2 feline sarcoma virus v abl insert. AB - Hardy-Zuckerman 2 feline sarcoma virus (HZ2-FeSV), isolated from a multicentric feline fibrosarcoma is a replication-defective acute transforming feline retrovirus which originated by transduction of feline c-abl sequences with feline leukemia virus (FeLV) and is known to encode a 110-kilodalton gag-abl fusion protein with tyrosine-specific protein kinase activity (P. Besmer, W. D. Hardy, E. E. Zuckerman, P. J. Bergold, L. Lederman, and H. W. Snyder, Nature (London) 303:825-828, 1983). The nucleotide sequence of the abl segment in the HZ2-FeSV genome was determined and compared with the murine and human v-abl and c-abl sequences. The predicted transforming protein consists of 344 amino acids (aa) of FeLV gag origin, 439 aa of abl origin, and at least 200 aa of FeLV pol origin (p110gag-abl-pol). The 1,317-base-pair HZ2-FeSV v-abl segment (fv-abl) corresponds to 5' abl sequences which include the region known to specify the protein kinase domain. The 5' 189 base pairs of fv-abl correspond to 5' c-abl sequences not contained in Abelson murine leukemia virus (MuLV) v-abl. The mouse c-abl exon which contains these segments was identified, and its nucleotide sequence was determined. Comparison of the predicted amino acid sequence of fv abl with those of Abelson MuLV v-abl and c-abl revealed five aa differences. The 5' junction between FeLV and abl was found to involve a preferred region in FeLV gag p30 (P. Besmer, J. E. Murphy, P. C. George, F. H. Qiu, P. J. Bergold, L. Lederman, H. W. Snyder, D. Brodeur, E. E. Zuckerman, and W. D. Hardy, Nature (London) 320:415-421, 1986). A six-base homology exists at the recombination site between the parental FeLV and the c-abl sequences. The 3' junction between fv-abl and FeLV pol predicts an in-frame fusion of fv-abl and FeLV pol. A transformed cell line containing a truncated gag-abl-pol protein, p85, that lacks most of the FeLV pol sequences was obtained by transfection of NIH 3T3 mouse cells. This result implies that the pol sequences of the p110gag-abl-pol protein are dispensable for fibroblast transformation. To assess whether the fv-abl segment specifies the unique biological properties of HZ2-FeSV, we constructed a Moloney MuLV-based version of HZ2-FeSV, Mo-MuLV(fv-abl), in which the fv-abl sequences were contained in a genetic context similar to that in HZ2-FeSV.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3029417 TI - Abundant expression of polyomavirus middle T antigen and dihydrofolate reductase in an adenovirus recombinant. AB - A modular gene with a cDNA encoding the polyomavirus middle T antigen positioned behind the adenovirus type 2 major late promoter and tripartite leader was substituted for the E1a region in an adenovirus vector. Permissive human cells infected with this recombinant produce middle T protein at levels as high as those of the most abundant late adenoviral proteins, e.g., hexon or fiber. This level represents at least a 40-fold increase over that observed in a polyomavirus lytic infection of murine cells. Partial proteolytic mapping showed that this protein has the same primary structure as middle T protein produced in polyomavirus-infected murine cells. The adenovirus recombinant-generated middle T protein exhibited in vitro kinase activity, although at an approximately 10-fold lower specific activity than that of middle T protein from polyomavirus-infected murine cells. Comparison of the expression levels of this middle T antigen containing adenovirus vector with a similar construction encoding dihydrofolate reductase suggested that the translation efficiency of the inserted gene was dependent upon the proximity of its initiation codon to the tripartite leader. We tested this possibility by comparing three dihydrofolate reductase recombinants among which the spacing between the initiation codon and tripartite leader varied from 188 to 36 nucleotides. The efficiency of expression of dihydrofolate reductase protein dramatically increased as this spacing was reduced. PMID- 3029416 TI - Genetic determinants of neoplastic diseases induced by a subgroup F avian leukosis virus. AB - Two subgroup F avian leukosis viruses, ring-necked pheasant virus (RPV) and RAV 61, were previously shown to induce a high incidence of a fatal proliferative disorder in the lungs of infected chickens. These lung lesions, termed angiosarcomas, appear rapidly (4 to 5 weeks after infection), show no evidence of proto-oncogene activation by proviral integration, and are not induced by avian leukosis viruses belonging to other subgroups. To identify the viral sequences responsible for induction of these tumors, we constructed recombinant viruses by exchanging genomic segments of molecularly cloned RPV with those of a subgroup A leukosis virus, UR2AV. The ability to induce rapid lung tumors segregated only with the env sequences of RPV; the long terminal repeat of RPV was not required. However, recombinants carrying both env and long terminal repeat sequences of RPV induced lung tumors with a shorter latency. In several cases, recombinant viruses exhibited pathogenic properties differing from those of either parental virus. Recombinants carrying the gag-pol region of RPV and the env gene of UR2AV induced a high incidence of a muscle lesion termed infiltrative intramuscular fibromatosis. One recombinant, EU-8, which carries the gag-pol and LTR sequences of RPV, and the env gene of UR2AV, induced lymphoid leukosis after an unusually short latent period. The median time of death from lymphoid leukosis was 6 to 7 weeks after infection with EU-8 compared with approximately 5 months for UR2AV. PMID- 3029419 TI - Complete interaction of cellular 56,000- and 32,000-Mr proteins with simian virus 40 small-t antigen in productively infected cells. AB - Two cellular proteins are found to be complexed with simian virus 40 small-t antigen in cellular extracts. The complex is a relatively unstable but dynamic one which can dissociate and reform in extracts. In extracts of permissive monkey kidney cells, the small-t antigen appeared to be present in excess, whereas the cellular proteins were nearly entirely committed to the complex in permissive monkey kidney cells. PMID- 3029421 TI - Immediate-early protein of pseudorabies virus is not continuously required to reinitiate transcription of induced genes. AB - We examined the role of herpesvirus immediate-early proteins in inducing and maintaining transcription by using tsG, a temperature-sensitive mutant in the immediate-early gene of pseudorabies virus. Cells infected at the permissive temperature were shifted to the nonpermissive temperature. Initially, early gene transcription rates were similar at both temperatures. Early gene transcription subsequently decreased slowly over several hours. Our results suggest that immediate-early protein function is required only for the initial activation of early gene transcription and is not required for multiple reinitiation events from activated early genes. PMID- 3029418 TI - Overproduction of polyomavirus middle T antigen in mammalian cells through the use of an adenovirus vector. AB - To overproduce biologically active polyomavirus middle T antigen, we used an adenovirus vector and human 293 cells as hosts. Two helper-independent recombinant adenoviruses were isolated that contain a hybrid transcription unit, in differing orientations, at a site in the adenovirus genome from which the E1a and most of the E1b transcription units have been deleted. The hybrid transcription unit consists of the adenovirus type 2 major late promoter and tripartite leader and a cDNA segment capable of encoding polyomavirus middle T antigen and accompanying 3' RNA-processing signals. Both recombinant viruses were stable and replicated to high titers in human 293 cells. The polyomavirus sequences were expressed, predominantly at late times after infection of 293 cells, to yield mRNAs that encoded middle T antigen. One of the recombinant viruses also expressed a middle T antigen-related protein in 293 cells. The latter was translated from one of several novel mRNA species that resulted from aberrant splicing and incomplete RNA processing of precursor RNA transcripts. Comparison of the amount of middle T antigen produced in 3T6 cells infected with polyomavirus with that in 293 cells infected with either of the recombinant adenoviruses, under optimal conditions for each system, revealed at least a 10 fold greater yield of the protein on a per-cell basis in the latter system than in the former. The recombinant-virus-encoded middle T antigen was biologically active, as evidenced by its ability to associate with and serve as a substrate for human pp60c-src. The functionality of the middle T antigen was further confirmed by demonstrating that both recombinant viruses efficiently transformed Rat-1 cells. These recombinant viruses will be useful to overproduce middle T antigen and to introduce the polyomavirus oncogene into a wide variety of mammalian cells. PMID- 3029420 TI - Mutational analysis of open reading frame E4 of bovine papillomavirus type 1. AB - Open reading frame (ORF) E4 is a 353-base-pair ORF of bovine papillomavirus type 1. To determine the biological activities of this ORF in mouse C127 cells, we analyzed the effects of two constructed mutations which are predicted to prevent synthesis of ORF E4 proteins while leaving the amino acid sequence encoded by the overlapping ORF E2 unchanged. Neither mutation interfered with the abilities of the mutants to efficiently induce focus formation, induce growth in soft agarose, or transactivate an inducible bovine papillomavirus type 1 enhancer. Also, neither mutation prevented establishment of the viral DNA as an extrachromosomal plasmid in transformed cells. These results suggest that ORF E4 proteins are not required for these biological activities, and they are consistent with the observation of others (J. Doorbar, D. Campbell, R. J. A. Grand, and P. H. Gallimore, EMBO J. 5:355-362, 1986) that the ORF E4 protein of a human papillomavirus is associated with late gene expression during papilloma formation. PMID- 3029422 TI - The 3' long terminal repeat of a transcribed yet defective endogenous retroviral sequence is a competent promoter of transcription. AB - Although actively transcribed and present as multiple genomic copies, a distinct class of endogenous murine leukemia virus-related sequence does not give rise to infectious virus. Since the long terminal repeat at the 3' terminus provides the transcriptional start site after reintegration, we determined the structure and potential promoter activity of that sequence obtained from cDNA of endogenous retroviral transcripts. These studies demonstrate that the distinctive 3' long terminal repeat sequence of these transcripts could serve as an effective promoter of transcription and, therefore, may not be the primary defect in the infectious cycle of retroviral replication but may result in the propagation of these endogenous retroviral sequences in the genome as retrotransposons. PMID- 3029423 TI - Course and extent of variation of equine infectious anemia virus during parallel persistent infections. AB - Comparisons of peptide and oligonucleotide maps of glycoproteins and RNA from nine isolates of equine infectious anemia virus (EIAV) that were generated during parallel infections of two Shetland ponies revealed that each isolate was structurally unique. Each EIAV isolate contained a unique subset of variant peptides, oligonucleotides, or both, indicating that structural variation in EIAV is a random and noncumulative process and that a large spectrum of possible EIAV variants can be generated in infected animals. PMID- 3029425 TI - Expression of the Rous sarcoma virus env gene from a simian virus 40 late-region replacement vector: effects of upstream initiation codons. AB - Expression of the Rous sarcoma virus envelope gene (env) from a simian virus 40 (SV40) late-region replacement vector is dependent on the position of env within the SV40 late-region sequences. The difference in expression levels appeared to be due to differences in the efficiency with which the env-specific transcripts were translated, because transcription levels from different constructions were similar. Deletion of the nucleotides encoding the agnoprotein initiator codon, located upstream of the env sequences in the poorly expressed construct, resulted in high levels of env expression. The agnoprotein initiator codon and overlapping open reading frame thus act as strong barriers to further ribosome scanning and prevent initiation at the env AUG codon. We conclude that AUG codons present in the late region of SV40 can reduce expression of inserted genes positioned downstream. Nevertheless, intrinsic properties of the gene may determine its ultimate level of expression. PMID- 3029424 TI - Mapping and sequencing of a gene from myxoma virus that is related to those encoding epidermal growth factor and transforming growth factor alpha. AB - Myxoma virus, a Leporipoxvirus and agent of myxomatosis, was shown to possess a gene with the potential to encode an epidermal growth factorlike factor. Its relationship to other members of this family, including the poxvirus growth factors from Shope fibroma virus and vaccinia virus, was analyzed. Alignment of DNA sequences and related open reading frames of myxoma virus and Shope fibroma virus indicated colinearity of genes between these poxviruses. PMID- 3029426 TI - Deletion mutations in the small t antigen gene alter the tissue specificity of tumors induced by simian virus 40. AB - Subcutaneous injection of wild-type simian virus 40 into Syrian hamsters normally induces fibrosarcomas at the injection site. We showed that subcutaneous injection of three different small t deletion mutants (dl884, dl883, and dl890) led to the formation of abdominal reticulum cell sarcomas (lymphomas) in about 15% of the animals bearing tumors. The remainder of the tumors were fibrosarcomas occurring with prolonged latencies at the site of injection. We postulated that, in the absence of an active small t protein, which is thought to have cell growth promoting properties, the mutant virus preferentially transforms rapidly proliferating lymphoid cells. PMID- 3029427 TI - Isolation and characterization of phosphonoacetic acid-resistant mutants of human cytomegalovirus. AB - Mutants of the human cytomegalovirus (HCMV) that were 6- to 13-fold more resistant to phosphonoacetic acid than the wild-type HCMV (Towne) were isolated. Extracts from mycoplasma-free, mutant-infected cells had phosphonoacetate resistant DNA polymerase activity in vitro. This strongly suggests that the selected mutations are in the HCMV DNA polymerase genes of these viruses. PMID- 3029428 TI - trans-activation of cellular and viral promoters by a transforming nonkaryophilic simian virus 40 large T antigen. AB - We used the chloramphenicol acetyl transferase (CAT) transient expression system to study the transactivating ability of a simian virus 40 (SV40) mutant that was unable to transport and localize large T antigen into the nucleus and which retained the competence to transform established cell lines. The effect of the SV40 wild type and the SV40 mutant for the large T antigen was tested in both mouse and simian cells on a series of plasmids in which the CAT gene was regulated by one of the following promoters: SV40 early and late, herpes simplex virus thymidine kinase, chicken alpha 2(I) collagen, mouse H-2Kk. Our results indicated that both the SV40 wild type and the cytoplasmic mutant for the large T antigen regulated transcription from any promoter tested, suggesting that the trans-activation by SV40 does not require the nuclear localization of the 100,000 molecular-weight large T-antigen protein. PMID- 3029429 TI - A bicistronic Epstein-Barr virus mRNA encodes two nuclear proteins in latently infected, growth-transformed lymphocytes. AB - EBNA2 is a nuclear protein expressed in all cells latently infected with and growth transformed by Epstein-Barr virus (EBV) infection (K. Hennessy and E. Kieff, Science 227:1230-1240, 1985). The nucleotide sequence of the EBNA2 mRNA (J. Sample, M. Hummel, D. Braun, M. Birkenbach, and E. Kieff, Proc. Natl. Acad. Sci. USA 83:5096-5100, 1986) revealed that it begins with a 924-base open reading frame that has an unusual potential translational initiation site (CAAATGG). This open reading frame is followed by 138 nucleotides with only one highly unlikely translational initiation site (TACATGC), which would translate a pentapeptide before the next stop codon. The last part of the mRNA is the open reading frame which encodes EBNA2. In this paper, we demonstrate that the 924-base open reading frame translates a 40-kilodalton protein in vitro or in murine cells transfected with the EBNA2 cDNA under control of the murine leukemia virus long terminal repeat. A protein of identical size was detected in EBV-transformed, latently infected human lymphocyte nuclei by using antibody specific for the leader open reading frame expressed in bacteria. Therefore, this is a rare example of a mRNA which translates two proteins from nonoverlapping open reading frames. Since the protein encoded by the leader of the EBNA mRNA is expressed in all nuclei of a latently infected cell line, it was designated EBNA-LP. EBNA-LP localizes to small intranuclear particles and differs in this respect from EBNA1, EBNA2, or EBNA3. EBNA-LP is not expressed in an EBV-transformed marmoset lymphocyte cell (B95-8) or in one EBV-infected Burkitt tumor cell line (Raji) but is expressed in three other Burkitt tumor cell lines (Namalwa, P3HR-1, and Daudi). PMID- 3029430 TI - Structural and transcriptional analysis of human papillomavirus type 16 sequences in cervical carcinoma cell lines. AB - We cloned and analyzed the integrated human papillomavirus type 16 (HPV-16) genomes that are present in the human cervical carcinoma cell lines SiHa and CaSki. The single HPV-16 genome in the SiHa line was cloned as a 10-kilobase (kb) HindIII fragment. Integration of the HPV-16 genome occurred at bases 3132 and 3384 with disruption of the E2 and E4 open reading frames (ORFs). An additional 52-base-pair deletion of HPV-16 sequences fused the E2 and E4 ORFs. the 5' portion of the disrupted E2 ORF terminated immediately in the contiguous human right-flanking sequences. Heteroduplex analysis of this cloned integrated viral genome with the prototype HPV-16 DNA revealed no other deletions, insertions, or rearrangements. DNA sequence analysis of the E1 ORF, however, revealed the presence of an additional guanine at nucleotide 1138, resulting in the fusion of the E1a and E1b ORFs into a single E1 ORF. Sequence analysis of the human flanking sequences revealed one-half of an Alu sequence at the left junction and a sequence highly homologous to the human O repeat in the right-flanking region. Analysis of the three most abundant BamHI clones from the CaSki line showed that these consisted of full-length, 7.9-kb HPV-16 DNA; a 6.5-kb genome resulting from a 1.4-kb deletion of the long control region; and a 10.5-kb clone generated by a 2.6-kb tandem repeat of the 3' early region. These HPV-16 genomes were arranged in the host chromosomes as head-to-tail, tandemly repeated arrays. Transcription analysis revealed expression of the HPV-16 genome in each of these two cervical carcinoma cell lines, albeit at significantly different levels. Preliminary mapping of the viral RNA with subgenomic strand-specific probes indicated that viral transcription appeared to be derived primarily from the E6 and E7 ORFs. PMID- 3029431 TI - Replication of adeno-associated virus in synchronized cells without the addition of a helper virus. AB - We investigated the helper-independent replication of adeno-associated virus (AAV) in cells synchronized by pretreatment with hydroxyurea, reversal of polyamine depletion, or physical mitotic detachment. In Chinese hamster cells (OD4 line) treated with hydroxyurea prior to infection. AAV underwent a complete cycle of replication. Transfection of such cells with plasmid-cloned AAV DNAs also gave rise to infectious viral progeny. Synchronization of OD4 cells by reversal of polyamine depletion or mitotic detachment led to independent AAV DNA synthesis (and infectious viral progeny in the case of the former procedure), but these procedures were not as effective as hydroxyurea pretreatment. Independent AAV DNA synthesis was also detected in some other cell lines of Chinese hamster, human, and monkey origin treated with hydroxyurea prior to infection. The results demonstrate that, in contrast to previous notions, the AAV infectious process is not absolutely dependent upon the addition of a coinfecting helper virus. PMID- 3029432 TI - Proteolysis of the p220 component of the cap-binding protein complex is not sufficient for complete inhibition of host cell protein synthesis after poliovirus infection. AB - Infection of cells with poliovirus results in the complete shutoff of host protein synthesis. It is presumed that proteolysis of the p220 component of the cap-binding protein complex that is required for the translation of host mRNAs is responsible for the shutoff phenomenon. In this paper, we show that when cells are infected with poliovirus in the presence of guanidine or 3-methylquercetin, both inhibitors of poliovirus replication, complete cleavage of p220 occurs by 3.5 h postinfection. However, under these conditions only 55 to 77% of host protein synthesis is suppressed. Results obtained with extracts prepared from poliovirus-infected cells were similar to those obtained in vivo. These results suggest that complete inhibition of host protein synthesis after poliovirus infection requires at least one event in addition to proteolysis of p220. Thus, proteolysis of p220 is probably necessary but not sufficient for total suppression of host protein synthesis after poliovirus infection. PMID- 3029433 TI - Characterization and nucleotide sequence of two herpes simplex virus 1 genes whose products modulate alpha-trans-inducing factor-dependent activation of alpha genes. AB - Herpes simplex viruses encode a structural protein which induces, in trans, expression of alpha genes, the first set of genes to be expressed after infection of permissive cells. This protein, designated as the alpha-trans-inducing factor (alpha-TIF), maps within the BamHI F fragment, and its gene has been sequenced. In the course of mapping the domain of the alpha-TIF gene, it was noted that the intact BamHI fragment was consistently more effective than the complete domain of the alpha-TIF gene in inducing expression of alpha genes. Cotransfections of DNA fragments containing an alpha indicator gene and the alpha-TIF gene with various regions of the BamHI F DNA fragment revealed that the sequences located 3' to the alpha-TIF gene raised the activity of the alpha-TIF gene to nearly the same level as that of the intact BamHI F fragment. The nucleotide sequence and S1 nuclease mapping analyses revealed the presence of two transcribed open reading frames capable of encoding polypeptides with translated molecular weights of 77,357 and 70,527. To determine whether the effect of these sequences in trans on alpha-TIF mediated induction of alpha genes was due to expression of these genes or competition for transcriptional factors, we constructed plasmids that contained both genes. Into each or both of these genes we inserted, near the translation initiation sites, 14-base-pair linkers carrying translational stop codons (TAG) in all three reading frames. Analyses of these plasmids indicated that the gene encoding the 70,527-molecular-weight polypeptide reduced alpha-TIF-dependent induction of alpha genes, whereas the gene encoding the 77,357-molecular-weight polypeptide increased this activity. Insertion of the stop codons abolished these activities. PMID- 3029434 TI - Endocrine and exocrine profiles of men with testicular tumors before orchiectomy. AB - In 15 patients with germ cell testicular tumors serum hormone profiles and semen analysis before orchiectomy were evaluated to determine the incidence of defective spermatogenesis associated with testicular tumors. Defective spermatogenesis was noted in 10 patients (66 per cent) on the basis of low sperm concentration, motility or semen volume. Of the 10 patients 7 had sperm concentrations less than 10 million per cc. Endocrine abnormalities occurred in 10 patients, the most common of which were elevations in serum human chorionic gonadotropin and estradiol, and a relative decrease in follicle-stimulating hormone. Three patients who presented with subfertile semen analyses were treated with orchiectomy alone. Repeat semen analyses 4 to 12 months after orchiectomy showed improvement in spermatogenesis and 2 patients achieved a normal semen analysis. Endocrine abnormalities and defective spermatogenesis are common in patients with testicular tumors. These abnormalities precede orchiectomy and imply that a primary germ cell defect exists in these patients. PMID- 3029435 TI - Indications for 99mtechnetium dimercapto-succinic acid scan in children. AB - The 99mtechnetium dimercapto-succinic acid scan provides an image of functional renal parenchyma. This static scan has specific indications and cannot be used simply in place of a 99mtechnetium diethylenetriaminepentaacetic acid scan. The major clinical indications for this investigation are the detection and/or evaluation of a renal scar, the small or absent kidney, an occult duplex system, certain renal masses, systemic hypertension or suspected vasculitis. The physiology of the 99mtechnetium dimercapto-succinic acid scan is reviewed briefly. PMID- 3029436 TI - Thymic enlargement following treatment for a metastatic germ cell tumor: a case report. AB - We describe a patient with metastatic testicular carcinoma in whom an anterior mediastinal mass developed 8 months after complete remission following chemotherapy and surgery. The mass was excised and pathological examination showed benign thymic hyperplasia. An anterior mediastinal mass encountered after successful chemotherapy does not always imply recurrence or relapse of malignant disease. The possibility of a benign etiology must be considered to prevent inappropriate treatment. PMID- 3029437 TI - Ketanserin interaction with urethral alpha-adrenoceptors. AB - The selective 5-HT2 receptor blocker ketanserin was found to reduce maximal urethral pressures in healthy females by about 40% without reducing blood pressure. In vitro, ketanserin completely or almost completely reduced contractions of the isolated female rabbit urethra induced by phenylephrine, noradrenaline (NA) and electrical field stimulation. The drug was less effective against responses evoked by clonidine and 5-hydroxytryptamine (5-HT). 5-HT induced contractions were effectively reduced by methysergide, but little affected by prazosin and rauwolscine. In concentrations exceeding 10(-7) M ketanserin significantly increased efflux of 3H in 3H-NA preloaded preparations of rabbit urethral muscle. Low concentrations of 5-HT, less than 10(-6) M, had slight inhibitory effects of 3H release, whereas 5-HT 10(-5) M caused a significant increase. It is concluded that ketanserin counteracts the effects of postjunctional alpha 1-adrenoceptor stimulation in isolated rabbit urethra. Such an effect might also explain its urethral pressure lowering action in man. PMID- 3029439 TI - Sealpox in captive grey seals (Halichoerus grypus) and their handlers. AB - Histopathologic, ultrastructural, and negative-staining studies indicated that nodular lesions on the flippers, head, and necks of recently weaned, captive grey seals (Halichoerus grypus) were similar to sealpox lesions reported from several other species of seals. Virions associated with the nodules were characteristic of the parapoxvirus subgroup of pox viruses. Two of the three persons handling the seals developed nodular lesions similar to "milker's nodules," the characteristic lesion in persons infected with parapoxvirus. The clinical course of the parapoxvirus infection in both the grey seals and their handlers is described. It was concluded that although sealpox is transmissible to man, the mild clinical manifestations place it in the nuisance category of zoonotic diseases. PMID- 3029438 TI - Effect of left atrial to left femoral artery bypass and renin-angiotensin system blockade on renal blood flow and function during and after thoracic aortic occlusion. AB - Temporary thoracic aortic occlusion can result in renal insufficiency with or without adjuncts to avoid distal hypoperfusion. In a canine model of thoracic aortic occlusion, left atrial to left femoral bypass was compared with blockade of the renin-angiotensin system. Renin-angiotensin system blockade with the converting enzyme inhibitor, MK422, resulted in restoration of baseline renal blood flow and glomerular filtration 30 minutes after cross-clamp release. Left atrial to left femoral bypass resulted in significant reduction in both renal blood flow and glomerular filtration 30 minutes after cross-clamp release. Stimulation of the renin-angiotensin system plays a significant role in the altered renal hemodynamics and glomerular filtration rates after thoracic aortic occlusion. PMID- 3029440 TI - Isolation of Powassan virus from a spotted skunk in California. PMID- 3029441 TI - Isolation of a retrovirus from the American bison and its relation to bovine retroviruses. AB - Tissue samples were removed at necropsy from five American bison (Bison bison) with clinical signs of a disease resembling malignant catarrhal fever (MCF). Using cell-associated virus techniques, attempts were made to isolate viruses from these tissues by culturing them directly or by co-culture with bovine fetal cells. Among the viruses isolated was one which was syncytiogenic and multiplied in bovine fetal spleen cells and remained highly cell-associated. The presence of reverse transcriptase activity indicated that it was a retrovirus. Also, it had antigenic cross activity with bovine syncytial virus, but not with bovine leukemia or bovine maedi-like retroviruses. We do not attribute a direct causative role of this retrovirus to MCF, but indirect relationships are possible. PMID- 3029443 TI - Diseases of wapiti utilizing cattle range in southwestern Alberta. AB - Specimens from 28 wapiti (Cervus elaphus canadensis) were collected by hunters in southwestern Alberta in 1984. Various tests were performed to detect infections and conditions that could affect cattle sharing the range or cause disease in wapiti. Serum antibodies were present against leptospiral serovars autumnalis (25%), bratislava (4%), and icterohaemorrhagiae (8%), and the viruses of bovine virus diarrhea (52%), infectious bovine rhinotracheitis (45%), and parainfluenza type 3 (13%). No serological evidence of bovine respiratory syncytial virus, Brucella, Anaplasma, bluetongue virus, or epizootic hemorrhagic disease virus was found, nor were any lesions of vesicular diseases, necrotic stomatitis or nutritional myopathy evident. Focal interstitial nephritis and sarcocystosis were diagnosed histologically in 40% and 75%, respectively, of the wapiti tested. The prevalence of giant liver flukes (Fascioloides magna) was 50% and of lungworms (Dictyocaulus viviparus) 32%. Leptospiral serology on cattle in the area did not indicate that wapiti or cattle were a serious source of infection to each other. The giant liver fluke was the parasite most likely to be amplified by wapiti for cattle. Within the limits of this study, the results indicated that wapiti in the Waterton area do not pose a disease threat to the cattle with which they range, but periodic observational studies in these wapiti would be a useful means of early detection of any changes in the interspecies relationship. PMID- 3029442 TI - Serologic survey for selected microbial pathogens of wolves in Alaska, 1975-1982. AB - Serum samples were collected from 116 wolves which were captured in southcentral Alaska during 1975 through 1982. Antibodies to the following infectious disease agents were found: infectious canine hepatitis virus-72 of 87 (81%), canine parvovirus type 2-0 of 55 (0%) through 1979 and 10 of 32 (31%) after 1979, Francisella tularensis-16 of 67 (25%), canine distemper virus-10 of 83 (12%), Coxiella burnetti-5 of 95 (5%), rabies virus-1 of 88 (1%), Brucella spp.-1 of 67 (1%), Leptospira interrogans-1 of 82 (1%). Apparently rabies, brucellosis, and leptospirosis were rare and had little effect on the wolf population. Conversely, the other five infections were comparatively common and may have had a negative impact on the health of specific individual wolves, but did not appear to influence the health of the population. PMID- 3029444 TI - Yoga or fiber? Thermodynamics in the vegetarian. PMID- 3029445 TI - Vaccine-associated paralytic poliomyelitis. United States: 1973 through 1984. AB - From 1973 through 1984, there were 138 cases of paralytic poliomyelitis reported in the United States; 105 (76%) were vaccine associated. Of the 105 vaccine associated cases, 35 occurred in recipients of oral polio vaccine (OPV), 50 in contacts to OPV recipients, 14 in immune deficient individuals, and six in individuals who had no history of receiving OPV or contact with recent OPV recipients. Thirty-three (94%) of the recipient cases, 41 (82%) of the contact cases, and five (36%) of the immune deficient cases were associated with the first dose of OPV. The overall frequency of vaccine-associated poliomyelitis was one case per 2.6 million doses distributed. However, the relative frequency of paralysis associated with the first dose in the OPV series was one case per 520,000 doses vs one case per 12.3 million subsequent doses. Vaccine-associated paralytic poliomyelitis is rare and the risks of OPV are small. The greatest likelihood of paralysis occurs in association with the first dose of OPV and that likelihood is reduced in subsequent doses more for recipients than for their contacts. PMID- 3029447 TI - Evidence for the implication of endogenous vasopressin in cardiovascular response to central alpha-adrenoceptor stimulation. AB - To determine the role of endogenous vasopressin (AVP) in cardiovascular response to central alpha-adrenoceptor stimulation, alpha 1-agonist methoxamine or alpha 2 agonist clonidine was administered intracerebroventricularly (ICV) to conscious Long-Evans (LE) rats as well as Brattleboro rats with hereditary hypothalamic diabetes insipidus (DI). In LE rats, ICV methoxamine increased blood pressure (BP) and decreased heart rate (HR), while ICV clonidine caused initial hypertension associated with bradycardia followed by prolonged hypotension with tachycardia. In DI rats, however, ICV methoxamine had no detectable effect on BP and HR, whereas ICV clonidine produced greater hypotension than in LE rats together with less initial bradycardia. Plasma levels of AVP increased 5-15 fold by methoxamine but did not change by clonidine. The intravenous (IV) but not ICV pretreatment with AVP vascular receptor antagonist d (CH2)5 Tyr (Me) AVP significantly attenuated the cardiovascular effects of methoxamine in LE rat, while neither IV nor ICV pretreatment with AVP antagonist modulated the cardiovascular effects of clonidine. These results provide the evidence for the implication of endogenous AVP in the cardiovascular response to central stimulation of alpha-adrenoceptors. PMID- 3029446 TI - Role of cyclic GMP of canine vascular smooth muscle in relaxation by organic nitrates. AB - The effects of organic nitrates on tone and tissue cyclic nucleotide levels were studied, using canine coronary, mesenteric and renal arteries, and femoral veins. Glyceryl trinitrate (GTN) relaxed all vascular tissues examined and increased tissue cyclic GMP (cGMP) levels in a concentration-dependent manner, but GTN induced no significant changes in cyclic AMP (cAMP) levels. An increase in cGMP levels induced by 10 microM of GTN in coronary arteries was observed before the onset of relaxation. Methylene blue, an inhibitor of guanylate cyclase, inhibited the relaxant effect of GTN and decreased cGMP levels. In contrast, M & B 22,948, an inhibitor of cGMP phosphodiesterase, not only enhanced relaxation by GTN, but also increased cGMP levels. Other organic nitrates, pentaerythritol tetranitrate (PETN), nicorandil (NIC), and isosorbide dinitrate (ISDN), also relaxed coronary arteries and increased cGMP levels in a concentration-dependent manner. A significant correlation was observed between percentage increases in cGMP levels and percentage relaxation by 10 microM of GTN, PETN, NIC, and ISDN (r = 0.952, p less than 0.001). Plasma concentrations of 4 organic nitrates inversely correlated with percentage increases in cGMP levels by 10 microM of these agents in coronary arteries (r = -0.845, p less than 0.001). These results suggest that an increase in cGMP is responsible for relaxation in vascular smooth muscles by organic nitrates, and that therapeutic plasma concentrations may be estimated by the degree of increase in cGMP levels induced by their administration. PMID- 3029448 TI - The in vivo cardiotoxic effect of eosinophilic cationic protein in an animal preparation. AB - We studied the in vivo effects of eosinophilic cationic protein (ECP) on DBA/2 mice, and compared the cardiac lesions caused by ECP with those caused by encephalomyocarditis (EMC) virus. ECP caused myocarditis in two of five mice (40%), and EMC virus did so in five of five mice (100%). Cardiac lesions of ECP were mild and limited to the right ventricular wall, which differed from those in the mice with EMC virus inoculation, where both the right and left ventricles were involved. This experiment is the first demonstration of the in vivo cardiotoxicity of ECP. PMID- 3029449 TI - Immunohistochemical study of the myocardium in murine Coxsackie B3 virus myocarditis using monoclonal antibodies--significance of Lyt 1 antigen-bearing lymphocytes in cell-mediated immunity. AB - This light- and electron-microscopic study using monoclonal antibodies provides an immunohistochemical demonstration of lymphocytes in situ in the myocardium in murine coxsackie B3 virus myocarditis. On the 7th and 9th days after virus inoculation, we observed many necrotic cardiocyte foci infiltrated with numerous inflammatory cells including macrophages and T lymphocytes. There were only a few Lyt 1-bearing lymphocytes in this phase of inoculation. On the 14th day there were many Lyt 1-bearing cells among the T cells in the foci. By immuno-electron microscopy, some Lyt 1-bearing T lymphocytes were seen in close contact with macrophages and/or other lymphocytes. These Lyt 1-bearing lymphocytes appeared to represent helper T cells, although Lyt 2.3+ lymphocytes were not examined. It is well known that activation of helper T cells leads to stimulation of B cells and cytotoxic T cells. It is assumed, therefore, that Lyt 1-bearing T lymphocytes that infiltrated into the myocardial foci play an important role in immune responsiveness at this stage of viral myocarditis. PMID- 3029451 TI - [Diltiazem potentiation of neuromuscular blockage produced by pancuronium or succinylcholine]. PMID- 3029450 TI - Myocarditis with myocardial infarction like findings in a 3-year old girl. AB - A 3 year-old Japanese girl had an acute onset associated with vomiting. The electrocardiogram (ECG) indicated changes similar to those of acute myocardial infarction (MI); there was no past history of kawasaki disease. Selective coronary angiography taken on the 28th day of illness revealed no abnormality. Thallium 201 scintigraphy was also performed and it revealed that the area of absent myocardial uptake was in the anterior wall. In serological findings, antibody titers against Coxsackie B-3 virus had risen significantly; therefore acute myocarditis caused by Coxsackie B-3 virus infection was diagnosed. PMID- 3029452 TI - Hemodynamic effects of BTB-cyclic AMP in anesthetized dogs. AB - The hemodynamic effects of 8-benzylthio-N6-n butyl adenosine 3',5' cyclic monophosphate (BTB-cAMP) were studied in anesthetized dogs. BTB-cAMP was given intravenously in doses of 1, 3 and 9 mg/kg, respectively. BTB-cAMP induced dose dependent increases in heart rate, max LV dp/dt, cardiac output and stroke volume, and decreases in mean blood pressure and total peripheral resistance. BTB cAMP had less of an effect on heart rate and blood pressure than dibutyryl cAMP (DB-cAMP), but showed a greater effect in augmenting cardiac contractility, with earlier manifestations of maximum action. Although the duration of its action is shorter, BTB-cAMP has possible therapeutic applications in cardiovascular diseases such as cardiogenic shock and congestive heart failure. It may also be a better agent than DB-cAMP for the treatment of patients with congestive heart failure because of a less potent hypotensive effect. PMID- 3029453 TI - [The influence of smoking on the development of various subtypes of pulmonary carcinoma]. AB - The influence of smoking on the development of pulmonary carcinoma of various histological subtypes was analyzed in 244 primary and 84 metastatic lung cancer patients. The new WHO classification was used for histological diagnosis. The relative risk was calculated by the odds ratio. It was found that all the squamous, small-and large-cell carcinomas were derived from smokers with relative risks of 27.8, 7.5 and 5.6, respectively. On the other hand, adenocarcinoma had very little, if any, relationship with smoking, the relative risk being 1.5 in men and 1.3 in women. The high-risk group for squamous cell carcinoma in men was defined with a regression line of y (duration of smoking) on x (number of cigarettes per day) as y = -0.24 x + 48.6. PMID- 3029454 TI - [Small-cell carcinoma of the lung relapsing at the primary site three years after radiation therapy with chemotherapy]. AB - A 55-year-old man was diagnosed as having limited disease (T2N2M0, stage III) of small-cell carcinoma of the lung. He underwent radiation therapy (60 Gy in 30 fractions) in addition to combination chemotherapy, resulting in complete response with advantage for irradiation. Relapse in the primary site was seen three years after completion of the initial treatment. As a result, retreatment was started using radiation therapy (50 Gy in 25 fractions) with chemotherapy. The tumor almost disappeared again. Subsequently, cerebral metastasis was observed in the fourth year, indicating clinical and prognostic significance. Thoracic irradiation contributed to prolonged survival in this case. The most effective manner of combining irradiation and chemotherapy and the most efficacious doses were discussed. PMID- 3029455 TI - [Classification and pathogenicity of retroviruses and HTLV-related viruses]. PMID- 3029456 TI - [AIDS and opportunistic infections]. PMID- 3029458 TI - [Viruses and autoimmunity]. PMID- 3029459 TI - [Metabolic bone diseases: various factors influencing bone resorption and osteogenesis--with special reference to the mechanism of action--parathyroid hormones and active-type vitamin D 3]. PMID- 3029457 TI - [Detection of adult T-cell leukemia virus and ATLV antibodies]. PMID- 3029460 TI - [Metabolic bone diseases: various factors influencing bone resorption and osteogenesis--with special reference to the mechanism of action--calcitonin]. PMID- 3029461 TI - [Metabolic bone diseases: hypophosphatemic vitamin-D resistant rickets]. PMID- 3029462 TI - [Metabolic bone diseases: vitamin-D-dependent rickets]. PMID- 3029463 TI - [Clinical application of beta-lactamase inhibitors on urinary tract infections]. PMID- 3029465 TI - [Clinical evaluation of serum CEA levels in cases of unresectable lung cancer]. PMID- 3029466 TI - [CT of intrahepatic cholangiocarcinoma]. PMID- 3029464 TI - [Thoracic CT: diagnosis of lymphatic metastasis of lung neoplasms]. PMID- 3029468 TI - [Diagnosis of hilar and mediastinal lymphadenopathy by MRI]. PMID- 3029467 TI - [Ultrasonographically guided percutaneous direct injection therapy of unresectable hepatoma]. PMID- 3029469 TI - [Immunohistological detection of antisera to casein in skin tumors]. PMID- 3029471 TI - [Prospective study of the development of hepatocellular carcinoma in 160 patients with cirrhosis of the liver]. PMID- 3029470 TI - Suppression or enhancement by superoxide dismutase of tumor cell killing by macrophages of normal donors and breast cancer patients. AB - This study was designed to examine the role superoxide production by macrophages plays in tumor killing. When superoxide dismutase was added to the normal macrophage-tumor cell (MA-160 cell line) suspensions macrophage mediated tumor cytotoxicity was suppressed. In contrast, superoxide dismutase often greatly enhances the cytotoxic ability of macrophages from breast cancer patients. To determine whether the inability of breast cancer patients' macrophages to kill tumor cells might be related to decreased levels of O2- production, we measured the amount of O2- generated by both normal and breast cancer patients' macrophages. The macrophages obtained from the normal donors produced 46.0 +/- 6.7 nanomoles O2-/10(6) macrophages per 10 minutes (mean +/- S.E. of 9 donors). In contrast, macrophages from the four breast cancer patients who possessed cytotoxic macrophages produced 56.4 +/- 5.2 nanomoles O2-/10(6) macrophages per 10 minutes whereas those from the 6 breast cancer patients who possessed non cytotoxic macrophages produced 18.6 +/- 3.2 nanomoles O2-/10(6) macrophages per 10 minutes. Thus, it appears that there is a relationship between macrophage cytotoxicity and the level of O2- produced. Furthermore, it is possible that the inability to generate sufficient amounts of O2- might explain the inability of breast cancer patients' macrophages to kill tumor cells in some cases. PMID- 3029472 TI - [Diagnosis and clinical features of portal vein tumor thrombosis in hepatocellular carcinoma]. PMID- 3029473 TI - [Serum prolyl hydroxylase values in liver diseases]. PMID- 3029474 TI - [Clinicopathological studies on 4 cases of mucin-producing adenocarcinomas of the pancreas]. PMID- 3029475 TI - [A case report of malignant glucagonoma with a profile of immunoreactive glucagon before and after the operation]. PMID- 3029476 TI - [Comparison of various therapeutic modalities with transcatheter arterial infusion of anticancer drugs in hepatocellular carcinoma]. PMID- 3029477 TI - [A case of plasma cell granuloma of the stomach combined with hepatocellular carcinoma]. PMID- 3029478 TI - [Regulation of prostaglandin production in cultured gastric mucosal cells]. PMID- 3029479 TI - Handy formula for calculating the probability of parentage exclusion. PMID- 3029480 TI - [Instrumentation physics in medicine: biomedical measurements using heavy particle beams]. PMID- 3029481 TI - [Epidemiological analysis of antibody to ATLA in chronic hemodialysis patients in Kumamoto Prefecture]. PMID- 3029482 TI - Immunohistochemical study on binding sites of synthetic rat-atrial natriuretic peptide in nephron-reduced rats. PMID- 3029483 TI - Effects of aldosterone and adrenocorticotropic hormone on urinary kallikrein excretion. PMID- 3029484 TI - Effect of calcitonin gene-related peptide on the cyclic AMP level of isolated mouse diaphragm. AB - The effect of calcitonin gene-related peptide (CGRP) on the cyclic nucleotide level in isolated mouse diaphragm was investigated. CGRP at concentrations of up to 1 microM caused dose-dependent increases in cyclic AMP levels but had no effect on cyclic GMP levels. At 1 microM, CGRP increased cyclic AMP levels by about 2.7-fold. Moreover, in the presence of phosphodiesterase inhibitor (Ro 20 1724), CGRP still caused even greater dose-dependent increases in cyclic AMP levels. Even in the presence of propranolol, a beta-adrenergic antagonist, CGRP stimulated increased cyclic AMP levels. In addition, specific binding of CGRP was observed in mouse diaphragm. All these results suggest that CGRP increases intracellular cyclic AMP levels via a CGRP receptor but not the beta-adrenergic receptor. PMID- 3029485 TI - Characteristics of ethacrynic acid highly sensitive Mg2+-ATPase in microsomal fractions of the rat brain: functional molecular size, inhibition by SITS and stimulation by Cl-. AB - Studies were performed to characterize ethacrynic acid (EA) highly sensitive Mg2+ ATPase isolated from microsomal fractions of the rat brain. The functional molecular sizes of the EA highly sensitive and EA less sensitive Mg2+-ATPases, estimated by a radiation inactivation method, were 480 and 80 kDa, respectively. An anion transport inhibitor, 4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonic acid (SITS) inhibited the EA highly sensitive Mg2+-ATPase activity. The type of inhibition was uncompetitive with respect to ATP, and the inhibition was suppressed by anions such as Cl-, Br- and I-. Chloride ions stimulated enzyme activity with an increase in Vmax, but not in Km, for ATP. Anions tested also increased the enzyme activity in the following order of decreasing potency: Cl- greater than Br- greater than CH3COO- = I- greater than SO4(2-) = HCO3- greater than SO3(2-). These results suggest that EA highly sensitive Mg2+-ATPase is a relatively large molecule with anion-sensitive sites that affect the ATP hydrolyzing activity and the SITS binding capacity through anions, with Cl- being the most potent. PMID- 3029487 TI - Evidence for the existence of stereoselective presynaptic beta 1-adrenoceptors on noradrenergic and dopaminergic neurons in the rat hypothalamus. AB - Using high performance liquid chromatography with an electrochemical detector, we examined antagonistic effects of l- and d-acebutolol and atenolol against isoproterenol-induced facilitation of impulse-evoked release of endogenous norepinephrine and dopamine in rat hypothalamic slices. Isoproterenol (10 nM) increased the release of the both catecholamines, and this facilitation was similarly antagonized by 100 nM l-acebutolol and 100 nM atenolol, but was not antagonized by 100 nM d-acebutolol. In conclusion, there exist stereoselective presynaptic beta 1-adrenoceptors on noradrenergic and dopaminergic neurons in the rat hypothalamus. PMID- 3029486 TI - Antidiuretic effects of dibutyryl-cyclic AMP microinjected into the hypothalamic supraoptic nucleus in water-loaded and ethanol-anesthetized rats. AB - Effects of dibutyryl-cyclic AMP (db-cAMP) and cyclic AMP (cAMP) microinjected into the hypothalamic supraoptic nucleus (SON) which contains the neurons synthesizing and releasing antidiuretic hormone upon the outflow and the osmotic pressure of urine and the other visceral functions were investigated in water loaded rats anesthetized with ethanol. When microinjected into the SON the dibutyryl analog of cAMP induced dose-dependent antidiuretic effects without significant effects on any other visceral functions. Dibutyryl-cAMP was much more effective than cAMP; The ED50 value for db-cAMP was approx. 200 nmol versus more than 500 nmol for cAMP. The time course of the antidiuretic effects was relatively slow with minimal urine outflow appearing only after more than 1/2 hour post-injection. The effects induced by db-cAMP demonstrated tachyphylaxis and were partially inhibited by pretreatment with atropine or theophylline, which suggests that the antidiuretic effects were mediated through muscarinic and adenosine receptors present in the nucleus. PMID- 3029489 TI - [Dermatomyositis and lung cancer]. PMID- 3029490 TI - [Pharmacological analysis of the function of the canine proximal urethra]. PMID- 3029488 TI - Active inflammatory change of the liver as a cause of postoperative hepatic failure. AB - Histopathology of cirrhosis was compared to the hepatic functional reserve and to the prognosis. One hundred and thirteen patients including 51 surgically treated for hepatocellular carcinoma (HCC) and 62 subjected to wedge biopsy of the liver during surgery for esophageal varices were studied. The type of cirrhosis associated with hepatocellular carcinoma was classified into 4 groups according to the degree of inflammation and the piecemeal necrosis. Of the 16 without cirrhosis, 13 (81 per cent) are living. Nine had an inactive cirrhosis and 5 (56 per cent) are living. Ten had a slightly active cirrhosis and 4 (40 per cent) are living. Sixteen had a fairly active cirrhosis and 7 (44 per cent) are alive. Immediate postoperative death due to acute hepatic failure occurred in 6, 4 of whom had a fairly active cirrhosis. In patients with active alcoholic hepatitis with numerous Mallory bodies and ballooning degeneration of liver cells, the prognosis was the poorest, if the liver was resected, even though the functional reserve seemed to be adequate. To prevent acute hepatic failure, liver histology during the surgery is predictive and wedge biopsy of the liver is recommended. PMID- 3029492 TI - Application of colorimetric assay for detection of tumor-associated antigens on bovine leukemic cells. PMID- 3029491 TI - Mechanism of suppression by bovine serum on growth of infectious bursal disease virus. PMID- 3029493 TI - Isolation of transmissible gastroenteritis virus from feces of diarrheic pigs in roller culture of CPK cells in the presence of trypsin. PMID- 3029494 TI - Preparation of monoclonal antibodies against Marek's disease virus and herpesvirus of turkeys. PMID- 3029495 TI - Widespread of parapox infection in wild Japanese serows, Capricornis crispus. PMID- 3029497 TI - Effects of vitamin A and dexamethasone on capsule collagen metabolism in mouse mammary adenocarcinoma. AB - It was previously shown that the administration of dexamethasone (Dex) to mice bearing mammary adenocarcinoma caused the collagen content of the tumor capsule to be decreased by 50%, but collagenase and other neutral protease activities of the tumor were the same as in the controls. These events occurred with distinctly increased tumor invasion and metastasis. The present communication reports on the characterization of capsule collagen and the effects of agents [vitamin A (VA) and Dex] on capsule collagen metabolism and presents further evidence concerning the possible mechanisms by which the collagen content of the capsule was decreased in the Dex-treated hosts. The collagen extracted from capsules of untreated controls and mice (C3H/HeJ) treated with VA or Dex was primarily type I, as judged by the migration of protein bands in sodium dodecylsulfate polyacrylamide gel electrophoresis and by patterns of elution peaks from an octadecyl C-18 column. The amino acid compositions of type I collagen of the capsule of treated and untreated controls were similar but not identical to those of mouse skin and guinea pig skin type I collagens. The specific activity of intracellular free [14C] proline, the extent of hydroxylation of [14C]proline residues of collagen, and the specific activity of [14C]hydroxyproline and proline in each case were similar in treated and in untreated controls. These results suggest that the observed 45% decrease in the conversion of [14C]proline into protein-bound [14C]hydroxyproline of the Dex-treated hosts apparently was due to decreased collagen synthesis. The data also suggest that the most critical effects of Dex on tumor invasion and metastasis appeared to occur at an early stage before full formation of the collagenous extracellular matrix. PMID- 3029496 TI - Polyadenylic acid metabolizing enzyme levels during induction of differentiation in a human leukemia T-cell line with phorbol ester. AB - Induction of differentiation of MOLT3 cells with 12-O-tetradecanoylphorbol-13 acetate (TPA) was found to affect the activity levels of the polyadenylic acid [poly(A)] metabolizing enzymes. TPA administration at a concentration of 16 nM resulted in an increase of poly(A) exonuclease and poly(A) polymerase activities after 2 and 2.5 hours, respectively, and in a decline to control levels thereafter. Cordycepin, an inhibitor of poly(A) polymerization, prevented the increasing binding of OKT11A monoclonal antibody induced by TPA treatment, whereas TPA-induced reduction in OKT6 monoclonal antibody binding persisted. The alterations in the poly(A) metabolizing enzyme activities following treatment of the cells with TPA, as well as the inhibitory effect of cordycepin, may suggest that these enzymes may be involved in the regulation of the differentiation process. PMID- 3029498 TI - The metabolism and mechanism of action of 1,25-dihydroxyvitamin D3. AB - Much has been learned about the formation of the active metabolite of vitamin D3, 1,25-dihydroxyvitamin D3. Information concerning its formation and catabolism has allowed a clear understanding of factors involved in the maintenance of plasma concentrations of the hormone. The effects of 1,25-dihydroxyvitamin D3 on calcium transporting cells in the intestine are marked and well defined. The tissue (intestinal tissue) is easily isolated and manipulated and hence, this is an ideal tissue in which to examine the mechanism of divalent cation transport. The mechanism by which 1,25-dihydroxyvitamin D3 brings about this effect should help in understanding sterol hormone action. PMID- 3029499 TI - Hydroxyl radical mediation of immune renal injury by desferrioxamine. AB - The acute phase of glomerular injury in a model of antiglomerular basement membrane, antibody-induced glomerulonephritis (antiGBM-GN) in rabbits was shown to be neutrophil-dependent using nitrogen mustard depletion studies. Administration of desferrioxamine (DFX) prevented the development of proteinuria in this model of renal injury [24 hr protein excretion (mean +/- SEM): antiGBM GN/DFX = 16.2 +/- 2.9 mg compared with antiGBM-GN control = 271.5 +/- 92.2 mg, P less than 0.01]. Antibody binding levels, glomerular filtration rates, circulating complement and neutrophil counts, glomerular C3 deposition, and neutrophil infiltration did not differ between DFX treated and antiGBM-GN groups. In vitro assay systems to assess oxygen radical production [superoxide anion (O2 ) and hydroxyl radical (OH.)] by neutrophils activated via the interaction of antiGBM antibody, GBM and complement were established. In these assays, DFX inhibited OH. production by immunologically-stimulated neutrophils (ISN) [nM diphenol/hr/10(6) cells, mean +/- SEM, ISN/DFX = 8 +/- 2 compared with ISN = 191 +/- 22, P less than 0.01] while production of O2- was not affected [nM O2 /hr/10(6) cells, mean +/- SEM, ISN/DFX = 29.1 +/- 4.3 compared with ISN = 32.6 +/ 2.5, P greater than 0.05]. These studies demonstrate that the iron chelator desferrioxamine can prevent neutrophil-dependent immune renal injury by interfering with neutrophil function. Treatment with the hydroxyl radical scavenger dimethylthiourea also significantly attenuated renal injury in antiGBM GN. Together, the in vivo and in vitro data strongly suggest that neutrophil dependent immunological renal injury is mediated via hydroxyl radical production by activated neutrophils within glomeruli. PMID- 3029500 TI - Electrophysiological indices of CNS function in hemodialysis and CAPD. AB - Fourteen Center hemodialysis (CHD) patients, 13 continuous ambulatory peritoneal dialysis (CAPD) patients, and 10 normals matched by group for age and sex were compared using a battery of evoked (EP) and event-related (ERP) brain potential measures to determine the effects on central nervous system (CNS) functioning of these two dialysis modes. While some differences were observed in early brainstem EP waves, the majority of differences occurred in the later waves associated with higher levels of cognitive processing. With tasks involving easy discriminations, CHD patients had longer latencies, indicative of less efficient cognitive processing, than CAPD patients who resembled controls. With difficult tasks, both CHD and CAPD groups showed abnormally-delayed later components. Similar results were obtained for the amplitude of the middle latency N1-P2 component, that is, reduced amplitudes for the CHD group with respect to both CAPD and control groups under low task demand, while both dialysis groups had reduced amplitudes under high task demand. These results suggest that CAPD patients are more similar to normals than CHD patients in ERP indices of attention and the efficiency of cognitive processing. PMID- 3029501 TI - Maternal contamination of some NZB sublines and genetic profiles of NZ-strains. AB - Six sublines of NZB mice bred in Japan were collected and their mitochondrial DNA (mtDNA) was examined by restriction analysis. The phenotypes of at least three of these sublines (NZB/Nrs, NZB/Nga and NZB/KlJms) differed from a standard one (NZB/BlWehi). Since mtDNA is inherited maternally, all sublines of a single inbred strain should share the same mtDNA phenotype. Therefore, b-type of mtDNA should be observed in all NZB sublines. Nevertheless, the above-mentioned sublines showed d-type mtDNA. These results suggested a genetic contamination of these sublines. This was confirmed by the finding that six aberrant alleles were detected also in their nuclear genomes using biochemical markers. For elucidation of the cause of contamination, we characterized the genetic profiles of four standard NZ-strains, NZB/BlWehi NZO/BlWehi, NZC/BlWehi and NZX/BlWehi, and of common inbred strains with black coat color, C57BL/6J, C57BL/10Sn, C57BL/Ks, C58/J and AU/SsJ. We found that five of the six aberrant alleles most strongly corresponded with those of C57BL/Ks. These results suggest that this contamination was ascribable to cross of NZB mice with a certain C56BL strain. We also deduced that NAB/BlPt and NZB/Full also probably were contaminated strains, suggesting that this contamination was not restricted to Japan. PMID- 3029502 TI - Diagnostic exercise: intermandibular swelling in rats. PMID- 3029504 TI - Beta-adrenergic regulation of secretion from Clara cell adenomas of the mouse lung. AB - Ethylnitrosourea-induced pulmonary adenomas of the mouse have been reported as being predominantly Clara cell in origin. The response of these tumor cells in vivo to the secretory agonist, isoproterenol (10 mg/kg) and the antagonist, propranolol (2.0 mg/kg) 1 hour after intraperitoneal injection into 120-day-old tumor-bearing mice was examined. Ultrastructural morphometry was used to quantitate the secretory response of tumor cells by measuring the volume density of the secretory granules. In the intact animal, isoproterenol stimulated secretion in the Clara cell adenomas (40% decrease in volume density with no change in surface to volume ratio of granules), while propranolol prevented this effect. In addition, beta-adrenergic receptors on isolated tumor cells were demonstrated by radioligand-binding assay by using [125I]iodocyanopindolol (ICYP). Scatchard analysis of data derived from whole cells indicates a maximum receptor-binding capacity of 27 fmoles/mg of protein and a KD of 0.029 nM. Isoproterenol displacement of ICYP binding yields an IC50 of 8 X 10(-7) M and a calculated KD of 3.36 X 10(-7) M. The beta 2 identity of these receptors was determined by utilizing the relatively specific beta 1 and beta 2 antagonists practolol and ICI-118,551, respectively. Practolol failed to displace more than 30% of ICYP binding even at 100 microM, while ICI-118,551 displacement of ICYP yielded a linear Hofstee plot (r = 0.93) and a KD of 5.04 X 10(-9) M. These findings suggest that the secretory activity of Clara cell-like pulmonary adenomas is under beta-adrenergic control similar to that of normal bronchiolar Clara cells. PMID- 3029505 TI - Detection and quantitation of urinary 11-nor-delta-9-tetrahydrocannabinol-9 carboxylic acid, a metabolite of tetrahydrocannabinol, by capillary gas chromatography and electron impact mass fragmentography. AB - A procedure for detection and quantitation of 11-nor-delta-9-tetrahydrocannabinol 9-carboxylic acid, a major metabolite of delta-9-tetrahydrocannabinol in urine, has been described. Since the metabolite is present in both conjugated and unconjugated forms, hydrolysis of urine was carried out to increase the sensitivity of detection. The acidic metabolite was isolated by strongly basic anion exchange resin, and subsequently derivatized to methyl 1-dehydroxy-1 methoxy-11-nor-delta-9-tetrahydrocannabinol-9-carbox ylate (1a) by methyliodide in the presence of tetramethylammonium hydroxide. The derivatized product was separated in a capillary column gas chromatograph, and finally detected by a mass spectrometer under electron impact mode. Confirmation of the product was carried out by monitoring three ions that represent the major portions of the molecule and comparing their relative abundances to that of a standard. Quantitation was based on 5'-2H3-11-nor-delta-9-tetrahydrocannabinol-9-carboxylic acid as internal standard. Excellent linearity was obtained over the range of 2 to 1000 ng/mL. The overall yield of extraction using the anion exchange resin was 50 to 60%. This extraction process is rapid and suitable for a large number of sample analyses. The methylated product (1a) is stable for at least 72 hr at room temperature. PMID- 3029506 TI - Prediction of blood lead by HPLC assay of erythrocyte pyrimidine 5'-nucleotidase. AB - A simplified preparation of erythrocytes (rbc) for rbc pyrimidine 5'-nucleotidase (P5N) is described and correlated with blood lead. Washed but unfiltered rbc are incubated with uridine monophosphate (UMP) and the amount of uridine liberated is measured by high performance liquid chromatography (HPLC). UMPase activity of unfiltered rbc of 20 normal subjects was 12.4 +/- 2.4 units, comparable to reported data obtained under varying conditions of preparation and P5N assay. In 20 lead-exposed adults and 20 controls, the regression coefficient for blood lead (PbB) predicted by rbc UMPase was In PbB = 4.77 - 0.22 UMPase; r2 = .61. If UMPase less than or equal to 7.6 units (mean, 2.0 SD) is considered abnormal, sensitivity for prediction of a blood lead greater than or equal to 40 micrograms/dL is 80%. Specificity, the percent of subjects with a UMPase greater than 7.6 units who had a blood lead less than 40 micrograms/dL, was 96%. Wider utilization of P5N, measured as UMPase, is proposed as a biologic correlate of blood lead. PMID- 3029507 TI - Cyanide in blood by gas chromatography with NP detector and acetonitrile as internal standard. Application on air accident fire victims. AB - An improved gas chromatographic method for HCN in blood with acetonitrile as an internal standard has been developed. The necessity to test for cyanide on samples from air accident fire victims is discussed in view of the fact that carbon monoxide measurements usually do not correlate with cyanide findings. Nine cases (out of 67), with blood CO saturation of less than 20% and cyanide concentration over 2 mg/L, are described. PMID- 3029503 TI - Tumor necrosis factor and interleukin 1: cytokines with multiple overlapping biological activities. PMID- 3029508 TI - Enhanced hepatic adenylate cyclase activity in rats with portacaval shunt. AB - Male Wistar rats were submitted to a portacaval anastomosis (PCA). Control rats were sham operated and pair fed. After 20 days, PCA led to a decrease in liver weight (-40%) and fasting blood glucose (-35%) and to an increase in fasting glucagonemia (+65%). The in vitro response of adenylate cyclase in hepatic membranes to GTP, Gpp(NH)p, fluoride, and forskolin (in the absence of GTP), and to glucagon (in the presence of GTP) was greater in PCA rats than in controls (by 30-54%) whereas the response to L-isoproterenol (in the presence of GTP) was only slightly increased (by 8%) and that to vasoactive intestinal peptide (in the presence of GTP) was similar in both groups of rats. The binding of [125I]glucagon and [125I]VIP to liver membranes did not differ in both groups of animals. It is concluded that the hepatic adenylate cyclase system from PCA rats responded better to stimuli involving efficiently the guanyl nucleotide stimulatory site Ns. This implies that the fasting hypoglycemia observed in these animals, in spite of the hyperglucagonemia, was due to either the refractoriness of a step distal to adenylate cyclase activation or to limited glucose production by an atrophic liver. PMID- 3029510 TI - Treatment and follow-up variables discriminating abstainers, controlled drinkers and relapsers. AB - A group of behaviorally treated abstainers, controlled drinkers and relapsers who had been followed up for a minimum of 1 year were compared on eight treatment and 11 posttreatment variables. Discriminating characteristics of abstainers were their goal of abstinence at discharge and length of attendance at aftercare facilities. Controlled drinkers had a goal of controlled drinking and were most likely to have had controlled drinking training during treatment. Relapsers were most likely to have been discharged from treatment prematurely, to receive further treatment and to use psychotropic medication. No controlled drinkers attended Alcoholics Anonymous or received residential care during follow-up in contrast to a small proportion of abstainers and relapsers who did. Some aspects of the drinking behavior of controlled drinkers and relapsers were also studied. Controlled drinkers were more likely to have a period of initial abstinence and for a longer period of time. Of the controlled drinkers, 66% drank at least once per week and 86% set themselves drinking limits; average consumption of alcohol was 48 and 32 g/day by men and women, respectively. The majority of controlled drinkers changed their preferred beverage and their social and physical drinking environment after treatment in accordance with therapeutic advice. PMID- 3029509 TI - Effects of the transmethylation inhibitor S-adenosyl-homocysteine and of the methyl donor S-adenosyl-methionine on rat Leydig cell function in vitro. AB - In purified rat Leydig cells, the methyl donor S-adenosyl-methionine (SAM), increases significantly in a dose dependent manner the [125I]hCG binding as well as the productions of cAMP and of testosterone; the competitive inhibitor of methylations S-adenosyl-homocysteine (SAH), has an opposite effect. Associated to oLH, SAM further enhances the cAMP synthesis while SAH inhibits significantly the adenylate cyclase activity. With regard to testosterone synthesis, SAM potentiates the stimulating roles of oLH and dbcAMP (27 and 38% increases, respectively) although SAH diminishes testosterone productions (48 and 35%, respectively under oLH and dbcAMP stimulations). Scatchard analysis has shown that SAM (1.4 mM) increases the number of LH/hCG binding sites on Leydig cells while SAH (1.4 mM) decreases it; LH/hCG Ka values are not modified neither by SAM nor by SAH. These data suggest that the in vitro regulation of steroidogenesis in purified rat Leydig cells may involve methylation processes (presumably phospholipids are the potential substrates of these reactions) which modulates the transmission of the hormonal signal through the membrane and affects the testosterone synthesis at a step beyond the adenylate cyclase. PMID- 3029512 TI - Hepatocellular carcinoma causing obstructive jaundice. AB - A 67-year-old man presented with signs and symptoms of obstructive jaundice. At autopsy, a hepatocellular carcinoma was noted to have obstructed both hepatic ducts and the common hepatic duct. Literature is reviewed to elaborate on this unusual manifestation of hepatocellular carcinoma. PMID- 3029513 TI - Oncocytic pleomorphic adenoma of the parotid gland. AB - A rare case of salivary gland pleomorphic adenoma, in which the majority of cells were oncocytic, is reported. The patient, a 53-year-old man, presented with a parotid gland mass that was otherwise asymptomatic. Surgical treatment consisted of a superficial parotid lobectomy. The specimen was prepared in standard fashion and studied by light and electron microscopy. The microscopic features were characteristic of salivary gland pleomorphic adenomas, however, both epithelial and "mesenchymal" elements were oncocytic. Such changes may occur focally in pleomorphic adenomas, but we were unable to find documentation of a wholly oncocytic variant in a review of the medical literature. From our clinical data and previous reports of pleomorphic adenomas with focal oncocytosis, it is concluded that such changes likely do not alter prognosis in affected patients. The possible significance of this lesion in regard to the histogenesis of salivary gland pleomorphic adenomas is discussed. PMID- 3029511 TI - Surgical treatment of bilateral synchronous Wilms' tumors. AB - Definite progress has been made in the treatment of bilateral Wilms' tumors with marked improvement in the prognosis. This is confirmed in our series of 6 consecutive patients with synchronous tumors. The recent trend toward more conservative surgery, double or triple drug chemotherapy, and avoidance of high dose radiation therapy has yielded good results. PMID- 3029514 TI - [Contribution of stereochemistry to the study of the spatial organization of pharmacological receptors]. AB - The important discovery by Pasteur of optical isomerism and the recent developments of stereochemistry showed that a complementarity exist between the geometry of molecules and their pharmacological receptors. The stereochemical bases and the principal configurational nomenclatures are briefly overviewed. The stereospecificity of the biological response and theories leading to an approach to stereochemical structures of main pharmacological receptors are developed. So, the biological activity of steroids is due to junctional modes of cycles and alpha or beta configurations of substituents. Acetylcholine has a skew conformation but it react by an anticlinal/anti-planar conformation with muscarinic receptor. To explain the difference in activity of adrenaline enantiomers, Easson and Stedman proposed a "three points" fixation to the adrenergic receptor. Dopaminergic receptor present a good degree of stereoselectivity: dopamine act by an anti-planar conformation in which the N-O distance is the same as in apomorphine (N-O10). The analgesic activity of morphinans is due to a cis junction of B and C cycles and to the stereoelectronic effect of the unshared lone pair on nitrogen. In the cyclamate sweeteners, some authors proposed for the sweet taste receptor a model with two points fixation (one acceptor and one donor) and two spatial barriers located at precise distances from this two sites. The stereoselectivity of molecules acting as substrates or inhibitors of enzymes is described. For example some oxazolidinone derivatives showed a selective inhibition toward monoamine oxidase A. Finally, the pharmacological activity falls often when molecules are administrated in racemic form. It seems that xenobiotics need to be dissymmetric for chiral recognition by biological systems. PMID- 3029515 TI - Selective preoperative evaluation for possible N2 disease in carcinoma of the lung. AB - The efficacy of computed tomography and surgical mediastinal exploration in determining tumor resectability were retrospectively evaluated in 92 consecutive patients with non-small cell lung carcinoma. Status of mediastinal nodes was ultimately determined by surgical mediastinal exploration or thoracotomy. Patients were divided into three groups on the basis of chest roentgenography: Group I comprised 30 patients with peripheral T1 or T2 lesions with normal hilar and mediastinal shadows. Only one patient was found to have an involved node. Chest roentgenography had an accuracy rate of 96% and computed tomography, 93%. Thoracotomy is recommended without either computed tomography or surgical mediastinal exploration in this group. Group II comprised 47 patients with T1 or T2 lesions with an abnormal hilus, an abnormal mediastinal shadow, or either the hilus or mediastinum obscured by overlying parenchymal disease. Computed tomography revealed mediastinal nodes 1 cm or greater in size (abnormal node group) in 21 patients (45%) and smaller than 1 cm (normal node group) in 26 patients (55%). Surgical mediastinal exploration was performed in the abnormal node group and involved nodes were found in 17 of 21 patients (81%). In the normal node group, thoracotomy only was performed and no involved nodes were found. Computed tomography is recommended in all patients in Group II. Patients in the normal node group may be subjected to thoracotomy only and those in the abnormal node group should undergo surgical mediastinal exploration as the next diagnostic step before thoracotomy. Group III comprised 15 patients with grossly abnormal mediastinal shadows. Findings from computed tomography were abnormal in all 10 patients in whom it was done. Surgical mediastinal exploration was done in all 15 and yielded abnormal results in 14. It is recommended in this group that computed tomography is unnecessary and surgical mediastinal exploration should be the only diagnostic procedure. Thus, in potentially resectable non-small cell lung carcinoma, the use of computed tomography and surgical mediastinal exploration should be selective and should be determined by appropriate initial interpretation of the chest roentgenogram. PMID- 3029516 TI - Role of magnesium and other divalent cations in ATP-utilizing enzymes. AB - While free MgATP shows tridentate coordination of Mg2+ by each of the three phosphates, enzyme-bound ATP generally coordinates metals at the beta- and gamma phosphates and only occasionally at the alpha-phosphate. Enzyme-catalyzed reactions of ATP are in-line nucleophilic displacements at either the alpha-, beta-, or gamma-phosphorus atom, as dictated by the relative positions of the enzyme-bound substrates. The divalent cation activates the attacked phosphoryl group and/or the leaving group of ATP by charge neutralization, electron withdrawal, and by adjustment of the conformation of the polyphosphate chain. Some ATP reactions require an additional divalent cation at the active site to adjust the protein conformation and/or to interact with the other substrate. PMID- 3029517 TI - Effect of magnesium depletion on 3H-ouabain binding site concentration in rat skeletal muscle. AB - In young (4-week-old) rats, dietary Mg depletion for 1.5 weeks was followed by a significant decrease in 3H-ouabain binding site or Na,K-pump concentration of up to 21% and in total K content of up to 11% in various hind limb muscles. K depletion was associated with a decrease in 3H-ouabain binding site concentration of up to 64% and in total K content of up to 31%. Mg depletion was associated with a similar correlation between the decrease in 3H-ouabain binding site concentration and total K content, as was K depletion (r = 0.98, p less than 0.001). Taken together, the results indicate that the decrease in the concentration of 3H-ouabain binding sites following Mg depletion is not primarily caused by Mg deficiency per se, but is probably secondary to the associated K deficiency. The decrease in Na,K-pump concentration may be of importance for the retardation of K repletion as well as the increase in digitalis toxicity observed with Mg deficiency. PMID- 3029518 TI - Classification of inhibitory amino acid receptors in the mammalian nervous system. AB - Electrophysiological and pharmacological evidence is summarized for the existence of an inhibitory receptor system operated by glycine and another two separate systems operated by gamma-aminobutyric acid (GABA) through GABA-A and GABA-B receptors, respectively. Claims for subclasses of GABA-A receptor are critically reviewed and found not-proven. A quantitative pharmacological profile of the GABA A receptor and associated regulatory sites for picrotoxin, barbiturates and benzodiazepines on the dorsal funiculus of the rat cuneate nucleus is described. When compared with this profile and the pharmacological properties of the glycine receptor complex, the effects of taurine cannot be entirely explained by actions on these two receptor systems. PMID- 3029520 TI - Evidence for the involvement of alpha-2 adrenoceptors in the sedation but not REM sleep inhibition by clonidine in the rat. AB - Rats with implanted electrodes for recording of EEG and EMG underwent 12-h recordings during the light period starting after i.p. injections of clonidine (0.1 mg/kg) alone or in combination with different alpha-adrenoceptor antagonists. Clonidine increased the proportion of time the rats spent in the drowsy stage of wakefulness which corresponds to behavioural sedation and inhibited both deep slow wave sleep and REM sleep for 6-9 hours. The amount of active wakefulness or light slow wave sleep were unaffected by clonidine. Yohimbine (1 mg/kg) reversed the increase in drowsy wakefulness by clonidine and increased active wakefulness without affecting sleep. Phentolamine (10 mg/kg) was ineffective against clonidine. Phenoxybenzamine (20 mg/kg) accentuated the sedative effect and prolonged the REM sleep inhibiting effect of clonidine. Prazosin (3 mg/kg) prolonged both the drowsy stage inducing and deep slow wave plus REM sleep inhibiting effects of clonidine. These electrophysiological results support the view that the sedative effect of clonidine in the rat is mediated by alpha-2 adrenoceptors, whereas in this species other mechanisms, possibly another population of alpha-2 receptors, may be involved in the clonidine-induced suppression of deep slow wave sleep and REM sleep. PMID- 3029519 TI - Tyrosine kinases in control of cell growth and transformation. AB - The growth of normal cells in tissues is strictly controlled, partly through intercellular communication by polypeptide growth factors. Malignantly transformed cells are independent from external growth factors to a certain extent, but their mechanisms for achieving growth autonomy differ from case to case. Several of the oncogene-encoded proteins are known to participate in the hormonal regulation of cell growth (for a recent review on tyrosine kinase oncogenes see ref. 21). Recent advances in molecular biology have shown important mechanisms for cell emancipation from growth regulatory hormonal signaling systems. A few such examples are discussed below. PMID- 3029521 TI - [Localization of 2 insulinomas by blood insulin gradients obtained by transparietal hepatic catheterization of the portal system]. PMID- 3029522 TI - [Lupus crisis versus cytomegalovirus infection. The role of digestive endoscopy in the differential diagnosis]. PMID- 3029523 TI - [Congenital and perinatal infections caused by viral agents, Toxoplasma gondii and Treponema pallidum. Study of 2000 cases and analysis of 488 positive cases]. PMID- 3029524 TI - [Occurrence of antibodies against hepatitis A virus in general population. Comparative study, 1977-1985]. PMID- 3029525 TI - Constipation in the cancer patient: causes and management. AB - Constipation is a distressing symptom with multiple etiologies in the cancer patient. Therapy is based on identification and management of underlying causes combined with symptomatic treatment. A working knowledge of bowel physiology and laxative pharmacology forms the foundation for this approach. PMID- 3029526 TI - Central cardiovascular effects of baclofen in spontaneously hypertensive rats. AB - This study was conducted to investigate the effects of centrally administered baclofen on blood pressure and heart rate in conscious spontaneously hypertensive (SHR) and normotensive Wistar-Kyoto (WKY) rats. Administration of baclofen (1.0 microgram/kg) into the lateral cerebral ventricle (icv) produced an increase in mean arterial pressure (MAP) in both SHR and WKY rats. The increase in MAP was significantly lower in SHR (13 +/- 3 mmHg) when compared with WKY (27 +/- 5 mmHg). The changes in heart rate (HR) were variable, from no change to a very small increase and did not differ significantly between SHR and WKY rats. The ability of baclofen to interfere with baroreceptor reflexes was also tested in separate experiments. In SHR, icv injection of baclofen (1.0 microgram/kg) significantly suppressed the pressor response and bradycardia evoked by phenylephrine 3.0 micrograms/kg iv, whereas in WKY, the pressor and HR responses to similar injections of phenylephrine were not affected by icv baclofen. Similarly, baclofen treatment modified hypotensive response and reflex tachycardia induced by nitroprusside (10.0 micrograms/kg) iv in SHR but not in WKY rats. Administration of sympathetic ganglionic blocker hexamethonium (HEX; 25 mg/kg) iv produced an equivalent decrease in MAP between SHR and WKY following icv injection of baclofen (1.0 microgram/kg). These results suggest that the effects of baclofen on the baroreceptor reflexes in SHR may not be mediated by a change in peripheral sympathetic tone. PMID- 3029527 TI - Corticosterone-releasing activity of immune mediators. AB - Products derived from the activated immune system have been reported to modulate neuroendocrine function. In addition, a direct connection between neuroendocrine and immune responses to stress has recently been proposed. We now provide evidence that heterogeneous lymphokine-containing supernatants from mitogen stimulated rat spleen cells can stimulate both basal and corticotropin-induced corticosterone secretion from rat adrenal cells in an in vitro perifusion system. Moreover, thymosin alpha 1, a 28-amino acid residue peptide found both in thymus and lymphocyte-derived supernatants was also able to synergistically stimulate corticotropin-stimulated corticosterone release, without affecting basal corticosterone output in this same in vitro adrenal cell perifusion system. These results reinforce the suggestion about the existence of bidirectional interactions between the immune and neuroendocrine systems. They also indicate that this communication may occur directly at the adrenal gland level, a major effector site of the body's response to stress. PMID- 3029528 TI - Effect of sodium ion on the affinity of naloxone for the kappa opioid receptor. AB - Several investigators have observed that sodium ion enhances the binding of naloxone to opioid receptors. This effect has generally been attributed to allosteric modulation of the state of the mu receptor. However, a recent claim has been made that the enhancement does not involve a change in the mu receptor, but instead occurs because naloxone becomes a more kappa-specific drug when sodium ion is present in high concentration. Since the claim was not based on experimental evidence from binding studies involving known high-affinity kappa ligands, we have investigated the competition of naloxone for the kappa site using [3H]U-69593 as the marker for receptor binding. Assays were carried out in the presence and absence of 100 mM NaCl. The results of the study indicate that sodium ion does not increase the affinity of naloxone or U-69593 for the kappa receptor. PMID- 3029529 TI - Effect of THIP and SL 76002, two clinically experimented GABA-mimetic compounds, on anterior pituitary GABA receptors and prolactin secretion in the rat. AB - In the present study, the ability of three direct GABA agonists, muscimol, THIP and SL 76002 to displace 3H-GABA binding from anterior pituitary and medio-basal hypothalamus membranes was evaluated. Further, the effect of both THIP and SL 76002 on baseline prolactin levels or after stimulation of hormone release with haloperidol has been also studied. Either muscimol, THIP or SL 76002 have shown to posses 7-, 7- and 3-fold higher affinity, respectively, for the central nervous system than for the anterior pituitary 3H-GABA binding sites. Moreover, THIP and SL 76002 have demonstrated to be respectively, 25- and 1000- fold less potent than muscimol in inhibiting 3H- GABA binding at the level of the anterior pituitary and about 25- and 2700- fold less potent at the level of the medio basal hypothalamus. Under basal conditions, either THIP or SL 76002 were ineffective to reduce prolactin release. However, after stimulation of prolactin secretion through blockade of the dopaminergic neurotransmission with haloperidol (0.1 mg/kg), both THIP (10 mg/kg) and SL 76002 (200 mg/kg) significantly counteracted the neuroleptic-induced prolactin rise with a potency which is in line with their ability to inhibit 3H-GABA binding in the anterior pituitary. The present results indicate that both compounds inhibit prolactin release under specific experimental situations probably through a GABAergic mechanism. In view of the endocrine effects of these GABA-mimetic compounds, the possibility arises for an application of these type of drugs in clinical neuroendocrinology. PMID- 3029530 TI - Effect of lecithin on the release of 5'-nucleotidase from liver plasma membrane of rat by bile acids. AB - Bile acids solubilize proteins from liver plasma membrane both in vivo and in vitro. The ability to solubilize the proteins is dependent on the species of bile acid. In this paper, the effect of phospholipid on the solubilization of a membrane-bound enzyme by bile acids was investigated in vitro. Taurocholate (TC) and tauroursodeoxycholate (TUDC) solubilized the enzyme, 5'-nucleotidase, from the liver plasma membrane of the rat in a concentration-dependent manner, although there was a great difference in their effect; at 40 mM, TC solubilized 55.4% of the original 5'-nucleotidase activity of the membrane, but TUDC only 5.7%. While lecithin alone had no solubilizing effect, its addition to the bile acids provoked a 10-fold increase in the solubilizing effect of TUDC, but virtually no change for TC, essentially equalizing the solubilizing effect of the two. Both TC-rich and TUDC-rich bile were obtained from rats infused with the respective bile acids via the jugular vein after their endogenous bile acid pool had been depleted. The solubilization effect of these biles was quite similar to the bile acid-lecithin mixtures. These findings demonstrate that lecithin enhances the ability of the bile acids to solubilize the membrane protein and eliminates the difference in the two bile acid species in their solubilizing ability. PMID- 3029531 TI - Anesthetic management of a patient with an undiagnosed panhypopituitarism. PMID- 3029532 TI - Endurance training and human alpha 2-adrenergic receptors on platelets. AB - Trained endurance athletes have a smaller rise in blood pressure, heart rate, and catecholamine response to stress when compared to an untrained, control population. Since pre-synaptic alpha 2-adrenergic receptors modulate the control of epinephrine release from nerve terminals, and since platelets are used as models of monoaminergic neurons, we investigated changes in the number and affinity binding of alpha 2-adrenergic receptors on platelets from endurance athletes and a sedentary population using the radiolabeled ligand yohimbine. We found, compared to a control, sedentary population, trained athletes had a 45% increase in the number of platelet alpha 2-adrenergic receptors (338 +/- 39 receptors/platelet vs 233 +/- 25 receptors/platelet, P less than 0.05) with no change in the dissociation constant (2.53 nM +/- 0.2 vs 2.24 nM +/- 0.2, NS) or the ED50 concentration for the competitive displacement of 2.5 nM yohimbine by epinephrine (2.8 X 10(-6) M vs 3.34 X 10(-6) M, NS). The increase in alpha 2 adrenergic receptors found in athletes may explain their decreased catecholamine response and concomitant physiologic responses to exertion. PMID- 3029533 TI - In vitro proton T1 and T2 studies on rat liver: analysis of multiexponential relaxation processes. AB - At 37 degrees C and 20 MHz, the T1 and T2 proton relaxation processes in intact rat liver tissue are multiexponential functions which in the majority of cases were decomposed into a major (alpha* approximately 90%, T1* = 374 ms, T2* = 58 ms) and a minor (alpha** approximately 10%, T1** = 130 ms, T2* = 181 ms) component. Both, T1 and T2, are temperature-dependent with a temperature shift of delta T1 = 1.5 ms/degrees C and delta T2 = 0.5 ms/degrees C, respectively. Storage of liver tissue at 4 degrees C and 37 degrees C led to remarkable changes of the T1 and T2 values. For T2 these changes occurred after a shorter storage time than for T1, but they are more pronounced for T1. To avoid such influences the relaxation measurements were performed within one hour after excision of the tissue. Even at 4 degrees C, long-term storage (greater than 3 h) must be avoided. A method for the quantitative determination of the fat content in liver based on multiexponential analysis of the T1 relaxation process was evaluated employing mixtures of triolein with liver homogenate. Triolein is a two-component system with T1* = 144 ms (alpha* = 62%) and T1** = 355 ms (alpha** = 38%). Finally, liver-specific protocol conditions were defined for in vitro relaxation studies. PMID- 3029534 TI - T1, T2, and relative proton density at 0.35 T for spleen, liver, adipose tissue, and vertebral body: normal values. AB - An MRI installation (Magnetom, Siemens, software version B1 of NUMARIS) working at 0.35 T was used to estimate T1, T2, and relative proton density in the spleen, liver, adipose tissue, and vertebral body in 14 healthy volunteers. Two double echo sequences were applied for all subjects: TR = 500 ms, TE1 = 35 and TE2 = 70 ms; and TR = 1600 ms, TE1 = 35 and TE2 = 70 ms. The images were sampled in regions of interest and appropriate relaxation expressions fitted to the ROI data yielding relaxation parameters and relative proton densities. Relaxation expressions, included in standard software (Siemens), were compared to more elaborate functions, developed in parallel to this study. The latter were found more appropriate, especially for high T1 values, and gave the following mean values for the four tissues (estimated uncertainty of mean in parentheses) T1 (ms) 915(36), 428(5), 261(7), and 501(11); T2 (ms) 79.7(8.8), 51.0(0.2), 59.8(1.0), and 64.7(0.8); and corresponding relative proton density (rho, arbitrary units) 2088(136), 2182(10), 2915(49), and 2136(21). The uncertainty in the values was estimated in the fitting procedure and does not include systematic errors. The relative noise in the ROIs was about 9% and the reproducibility of the ROI mean values about 8%. PMID- 3029536 TI - Methods for assessing exocytosis by neutrophil leukocytes. PMID- 3029535 TI - Measurement of solute proton spin-lattice relaxation times in water using the 1,3,3,1 sequence. AB - 1H NMR spin-lattice relaxation times (T1) of the N-CH3 proton resonances of phosphocreatine (PCr) and creatine (Cr) in water solutions were obtained using the 1,3,3,1 pulse sequence. These T1 values were equivalent to those obtained in D2O and water using either the conventional inversion-recovery experiment or the 1,3,3,1 pulse sequence. Thus, the 1,3,3,1 sequence of proton NMR can provide an independent means along with phosphorous NMR for assess PCr and for the study of the creatine kinase reaction (PCr + ADP in equilibrium ATP + Cr) in aqueous solutions and perhaps in biological preparations. PMID- 3029537 TI - Respiratory burst during phagocytosis: an overview. PMID- 3029538 TI - Human myeloperoxidase and hemi-myeloperoxidase. PMID- 3029539 TI - Microassays for superoxide and hydrogen peroxide production and nitroblue tetrazolium reduction using an enzyme immunoassay microplate reader. PMID- 3029540 TI - Measurements of cyclic adenosine monophosphate and cyclic guanosine monophosphate levels in polymorphonuclear leukocytes, macrophages, and lymphocytes. PMID- 3029541 TI - Cytotoxicity by natural killer cells: analysis of large granular lymphocytes. PMID- 3029542 TI - Determination of antibiotic activities with the aid of luminous bacteria. PMID- 3029544 TI - Purification and characterization of Chlamydomonas flagellar dyneins. PMID- 3029543 TI - Isolation of fascin, an actin-bundling protein, and SU45, an actin severing/capping protein from sea urchin eggs. PMID- 3029546 TI - Preparation of a fluorescent analog: acetamidofluoresceinyl-labeled Dictyostelium discoideum alpha-actinin. PMID- 3029545 TI - Preparation and assay of cytoplasmic and secreted gelsolin. PMID- 3029547 TI - Isolation of metavinculin from chicken smooth muscle. PMID- 3029548 TI - Apocytochrome c induces pH-dependent vesicle fusion. AB - The ability of apocytochrome c and the heme containing respiratory chain component, cytochrome c, to induce fusion of phosphatidylcholine (PC) small unilamellar vesicles containing 0-50 mol % negatively charged lipids was examined. Both molecules mediated fusion of phosphatidylserine (PS):PC 1:1 vesicles as measured by energy transfer changes between fluorescent lipid probes in a concentration- and pH-dependent manner, although cytochrome c was less potent and interacted over a more limited pH range than the apocytochrome c. Maximal fusion occurred at pH 3, far below the pKa of the 19 lysine groups contained in the protein (pI = 10.5). A similar pH dependence was observed for vesicles containing 50 mol % cardiolipin (CL), phosphatidylglycerol (PG), and phosphatidylinositol (PI) in PC but the apparent pKa values varied somewhat. In the absence of vesicles, the secondary structure of apocytochrome c was unchanged over this pH range, but in the presence of negatively charged vesicles, the polypeptide underwent a marked conformational change from random coil to alpha helix. By comparing the pH dependencies of fusion induced by poly-L-lysine and apocytochrome c, we concluded that the pH dependence derived from changes in the net charge on both the vesicles and apocytochrome c. Aggregation could occur under conditions where fusion was imperceptible. Fusion increased with increasing mole ratio of PS. Apocytochrome c did induce some fusion of vesicles composed only of PC with a maximum effect at pH 4. Biosynthesis of cytochrome c involves translocation of apocytochrome c from the cytosol across the outer mitochondrial membrane to the outer mitochondrial space where the heme group is attached. The ability of apocytochrome c to induce fusion of both PS-containing and PC-only vesicles may reflect characteristics of protein/membrane interaction that pertain to its biological translocation. PMID- 3029549 TI - Correlation between the number of placental opioid receptors, mode of delivery, and maternal narcotic use. AB - Human placental opioid receptors were assayed using the radioactive opioid agonist, etorphine, to determine the number of binding sites in villous tissue membrane preparations. Significant differences in receptor concentration per milligram of protein of tissue were found between placentas obtained following vaginal or abdominal delivery (P less than 0.002). Labor itself did not alter apparent receptor numbers. In patients with maternal narcotic abuse during pregnancy, no opioid binding could be detected regardless of the mode of delivery, suggesting possible receptor down-regulation. PMID- 3029550 TI - [Peroxidase of propionic acid bacteria]. AB - The peroxidase activity was found in Propionibacterium shermanii. Methods were developed to isolate and purify the enzyme. It was shown to be a heme-containing protein, specific to H2O2, stable at 20 to 30 degrees C and exerting the optimal action at pH 6.8 to 7.0. The rate of the enzyme-catalysed reaction was studied as a function of the enzyme and substrate concentrations. The Km was determined for H2O2 and o-dianisidine. PMID- 3029551 TI - [Mechanism of growth limitations in a recombinant strain of Escherichia coli]. AB - A recombinant Escherichia coli strain with the cloned gene of restriction endonuclease EcoR-V was studied. The diauxic growth of this strain under batch conditions, the composition of excreted products and the hyperbolic shape of the chemostat growth curve has made it possible to formulate the following hypothesis. At a high flow rate, one of the first reactions in the tricarboxylic acid cycle or a reaction directly preceding the cycle is a limiting step in the metabolism of E. coli cells. The saturation of the limiting reaction with a substrate at a flow rate higher than the critical one may be responsible for the hyperbolic profile of chemostat curves. The above hypothesis was confirmed by experiments conducted under extreme conditions, in which the composition of the growth medium was changed and the temperature was raised so that the cloned protein synthesis was depressed. Under such conditions, the enzyme composition of the cell was fixed and determined by the physiological state of the culture at a time when the temperature was elevated. The hypothesis was also supported by the results obtained in growing the strain with the induced protein synthesis in media to which various compounds were added. PMID- 3029552 TI - [Cleavage of DNA from 2 phages of Bacillus thuringiensis by EcoRI and HindIII endonucleases]. AB - The DNA of two Bacillus thuringiensis phages was restricted by endonucleases EcoRI and HindIII and the electrophoretic distribution of the fragments in agarose gel was studied. EcoRI was shown to restrict the DNA of phage 1-97A into 8 fragments and the DNA of phage 1-97B into 12 fragments. Restriction with HindIII results in the formation of 22 and 9 fragments for phage 1-97A and phage 1-97B, respectively. The molecular mass of the DNAs determined by summing up EcoRI restricts is 80.87 MDa for phage 1-97A and 32.45 MDa for phage 1-97B. PMID- 3029553 TI - Vasodilatation and the discovery of endothelium-derived relaxing factor. AB - Whether endothelium-derived relaxing factor (EDRF) is important in the physiology and pathology of vascular reactivity must await the discovery of the nature of the factor and an appropriate antagonist substance. Nevertheless the demonstration that EDRF is powerful, short-lived and can be released by a wide variety of stimuli adds a new dimension to our knowledge of the control of the circulation. The macroenvironment of the vessel wall is influenced by nerves, blood-borne factors and the architecture of the wall. The discovery of a role of the endothelial cell has for the first time forced pharmacologists to consider cell-cell interactions in the micro-environment of the blood-vessel wall. This is a potentially important target for new therapy. PMID- 3029554 TI - Monocyte and iscom enhancement of cell-mediated response to cytomegalovirus. AB - Cell-mediated and humoral responses to cytomegalovirus were studied in a monkey model. Repeated low doses of virus antigen gave poor reactivities in both respects. High antigen doses gave a good humoral IgG response. When autologous monocytes were incubated with the CMV antigen as the immunizing injection, the specific cellular response to CMV antigen increased. The monocytes themselves did not contribute to the in vitro specific proliferation response. When iscoms were the carrier particles for CMV antigens, cellular response was even more strongly enhanced. In immunization schedules where specific cellular responses are important, we suggest that autologous monocytes or iscoms may be employed as antigen carriers. PMID- 3029556 TI - [Experimental evidence for infection of Culex pipiens L. mosquitoes by West Nile fever virus from Rana ridibunda Pallas and its transmission by bites]. PMID- 3029555 TI - Study of the chemical nature of Frp/3 cell recognition units for hepatitis A virus. AB - Research has been carried out in order to clarify the chemical nature of cell receptors interacting with a fast growing strain of hepatitis A virus (HAV) producing a cytopathic effect on Frp/3 cells. Cell surface susceptibility to HAV attachment has been studied after treatment with enzymes acting on different chemical groupings. Results obtained showed a lowering of cell susceptibility to HAV infection following the action of phospholipase A2, phospholipase C, trypsin and beta-galactosidase. These data suggested that phospholipids, proteins and galactose participate to the cellular receptorial area for HAV. PMID- 3029557 TI - [Primary malignant fibrous histiocytoma of the breast, report of a case]. AB - Although malignant fibrous histiocytoma (MFH) is the most common soft tissue sarcoma of adult, it rarely arises in the breast tissue. Only seven cases have been previously reported in Japan. One case of the MFH of the mammary gland is presented in this paper. A 39 year old woman, who had received no radiation, was referred to our hospital due to a painful right breast lump. Excisional biopsy revealed yellowish white soft tumor with an obscured margin. Histological, tumor was composed of spindle shaped cells of a storiform pattern, which was diagnosed as MFH of the breast. Radical mastectomy (Br + Mj + Ax) was added to this patient. The tumor invaded to the fat tissue without metastasis to the axillary lymph nodes. We performed the post-operative adjuvant chemotherapy of Adriamycin. She made an uneventful recovery and is now free of disease eleven months postoperatively. PMID- 3029558 TI - Glucose phosphotransferase and intracellular trafficking. AB - Glycoproteins containing phosphodiester-linked glucose residues have recently been described. The synthesis of this structure occurs due to the intact transfer of alpha glucose-1-phosphate from UDP-glucose and is catalyzed by the enzyme glucose phosphotransferase (GlcPTase). The endogenous acceptors for GlcPTase have been characterized as to molecular weight following incubation of selected homogenates with (beta 32P)UDP-glucose. These glycoproteins are distinct from the lysosomal hydrolases recognized by the GlcNAc phosphotransferase. The transfer of 32P from (beta 32P)UDP-Glc can also be detected when the nucleotide sugar is microinjected into the cytoplasm of individual neurons in Aplysia. The phosphorylated acceptors in this system seem to be predominantly two glycoproteins that are subjected to rapid axoplasmic transport. The possible role of this post-translational modification in the intracellular trafficking of a subset of newly synthesized glycoproteins is discussed. PMID- 3029559 TI - Lymphocyte attachment to high endothelial venules during recirculation: a possible role for carbohydrates as recognition determinants. AB - During the course of their recirculation through the body, blood-borne lymphocytes specifically adhere to high endothelial venules (HEV) within secondary lymphoid organs such as peripheral lymph nodes (PN) and gut-associated Peyer's patches (PP). This adherence event, which initiates the extravasation of the lymphocyte, is highly specific in terms of the class of lymphocyte and the anatomic location of the HEV. We review evidence that the lymphocyte adhesive molecule ('homing receptor') involved in attachment to PN HEV is a carbohydrate binding receptor (lectin-like) with specificity for mannose-6-phosphate (M6P) like ligands. We describe the use of a novel cytochemical probe for the detection and characterization of cell surface carbohydrate-binding receptors. Using a M6P based probe, we show that the carbohydrate-binding receptor on lymphocytes is closely-related or identical to the MEL-14 antigen, a putative homing receptor identified by a monoclonal antibody. Evidence is presented that the lymphocyte attachment sites on both PN and PP HEV are inactivated by mild periodate oxidation and hence are probably carbohydrate in nature. Yet, the sites are biochemically distinguishable in that one class (PN) requires sialidase-sensitive structures whereas the other (PP) does not. We raise the possibility that diversity in the carbohydrate-based recognition determinants on HEV may underlie the adhesive specificities in this system. PMID- 3029561 TI - Augmentation of c-fos mRNA expression by activators of protein kinase C in fresh, terminally differentiated resting macrophages. AB - Expression of c-fos mRNA was investigated in fresh, normal peritoneal macrophages (M phi), which are terminally differentiated, nonproliferating cells. The levels of c-fos mRNA were dramatically increased by stimulation with phorbol myristate acetate (PMA), calcium ionophore, or 1-oleoyl-2-acetoyl glycerol (OAG). Induction of c-fos mRNA by all the above agents followed similar kinetics, with a peak of mRNA 30 min after stimulation. These results demonstrate that c-fos mRNA can be augmented in fresh, terminally differentiated cells. Since the stimuli increasing c-fos mRNA are direct or indirect activators of protein kinase C, our data suggest that in M phi c-fos mRNA is controlled by protein kinase C activation. PMA, calcium ionophore, and OAG were biologically active in M phi. PMA and calcium ionophore induced respiratory burst and tumoricidal activity, respectively, whereas OAG and PMA were chemotactic for M phi. Interferons beta and gamma, potent M phi activators eliciting tumoricidal activity, did not alter the levels of c-fos mRNA. These results indicate that c-fos mRNA augmentation is a stimulus-specific rather than a function-specific response connected to activation of protein kinase C. PMID- 3029562 TI - Physical analysis of the COR region: a cluster of six genes in Saccharomyces cerevisiae. AB - Six genes, CYC1, UTR1, UTR3, OSM1, tRNAGly, and RAD7, have been localized within an 8-kilobase region on chromosome X of the yeast Saccharomyces cerevisiae. The physical structures and the transcripts of these genes were identified by analyzing a normal strain and six deletion mutants by genomic blotting, transcriptional analysis, and gene disruption procedures. The well-studied CYC1 gene encodes iso-1-cytochrome c; the tRNAGly gene encodes a tRNA; deletion of OSM1 and RAD7 causes sensitivity to hypertonic medium and UV irradiation, respectively. There were no observable phenotypes in strains having deletions of the UTR1, UTR3, and tRNAGly gene. The high density of transcripts, with little or almost no intragenic regions, indicates that the chromosomal organization of S. cerevisiae resembles the chromosomal organization of procaryotes rather than higher eucaryotes. PMID- 3029563 TI - Activation of the ribosomal DNA promoter in cells exposed to insulinlike growth factor I. AB - We constructed a stable cell line, 3T3A5, which carried a chimeric gene in which the simian virus 40 T-antigen-coding gene was under the control of the mouse ribosomal DNA promoter. These cells expressed T antigen when they were growing exponentially in 10% fetal calf serum, but they all became T negative when incubated for 5 days in low-concentration serum. The readdition of serum or platelet-poor plasma again induced the expression of T antigen, which was accompanied by an increase in steady-state levels of the corresponding RNA. Among the various growth factors tested for their ability to induce T-antigen expression in 3T3A5 cells, only insulinlike growth factor I (IGF-I) could induce T antigen at physiological concentrations. The effect of IGF-I or platelet-poor plasma was abolished by an antibody to IGF-I. Other growth factors, like insulin and epidermal growth factor, could induce the expression of T antigen in 3T3A5 cells, but only at concentrations far above the physiological range. Other growth factors were totally ineffective. These results indicate that exposure of cells to IGF-I can activate transcription from the ribosomal DNA promoter. PMID- 3029560 TI - Current ideas on the significance of protein glycosylation. AB - Carbohydrate has been removed from a number of glycoproteins without major effect on the structure or enzyme activity of the protein. Thus carbohydrate has been suggested to underly a non-primary function for proteins, such as in relatively non-specific interactions with other carbohydrates or macromolecules, stabilization of protein conformation, or protection from proteolysis. This non specific concept is consistent with both the general similarity in carbohydrate structure on very diverse glycoproteins and the frequent structural microheterogeneity of carbohydrate chains at given sites. The concept is supported in a general sense by the viability of cells whose glycosylation processes have been globally disrupted by mutation or pharmacological inhibitors. In contrast to the above observations, other studies have revealed the existence of specific, selective receptors for discrete oligosaccharide structures on glycoproteins which seem to be important for compartmentalization of the glycoprotein, or the positioning of cells on which the glycoprotein is concentrated. Sometimes multivalency in the carbohydrate-receptor interaction is crucial. There are additional possible roles for carbohydrate in the transduction of information upon binding to a receptor. The possibility of specific roles for carbohydrate is supported by the existence of numerous unique carbohydrate structures, many of which have been detected as glycoantigens by monoclonal antibodies, with unique distributions in developing and differentiated cells. This article attempts to summarize and rationalize the contradictory results. It appears that in general carbohydrate does in fact underlie only roles secondary to a protein's primary function. These secondary roles are simple non-specific ones of protection and stabilization, but often also satisfy the more sophisticated needs of spatial position control and compartmentalization in multicellular eukaryotic organisms. It is suggested that there are advantages, evolutionarily speaking, for the shared use of carbohydrate for non-specific roles and for specific roles primarily as luxury functions to be executed during the processes of cell differentiation and morphogenesis. PMID- 3029564 TI - The SPT6 gene is essential for growth and is required for delta-mediated transcription in Saccharomyces cerevisiae. AB - Mutations in the Saccharomyces cerevisiae SPT6 gene were originally identified as one class of extragenic suppressors of Ty and delta insertion mutations in the 5' noncoding regions of HIS4 and LYS2. We cloned SPT6 and constructed a null allele by gene disruption. Haploid spores carrying the spt6 null allele were inviable, indicating that the SPT6 gene is essential for mitotic growth. SPT6 was mapped to the right arm of chromosome VII, 44 centimorgans (cM) from ADE6 and 9 cM from CLY8. We showed that spt6 mutations suppress delta insertion mutations at the level of transcription but have no qualitative or quantitative effect on Ty transcription. In addition, we observed interesting SPT6 gene dosage effects. An SPT6 strain containing a high-copy-number plasmid clone of SPT6 showed suppression of delta insertion mutations, and a diploid strain with half its normal dose of SPT6 (SPT6/spt6 null) also exhibited suppression of delta insertion mutations. Therefore, having either too many or too few copies of SPT6 causes a mutant phenotype. Finally, this study and that in the accompanying paper (L. Neigeborn, J. L. Celenza, and M. Carlson, Mol. Cell. Biol. 7:679-686, 1986) showed that spt6 and ssn20 mutations (isolated as suppressors of snf2 and snf5 [sucrose nonfermenting] mutations) identify the same gene. SPT6 and SSN20 have the same genetic map position and share an identical restriction map. Furthermore, spt6 and ssn20 mutations fail to complement each other, and ssn20 mutations suppress solo delta insertion mutations at HIS4 and LYS2. These results, taken in conjunction with the SPT6 dosage effects and the fact that SPT6 is an essential gene, suggest that SPT6 plays a fundamental role in cellular transcription, perhaps by interaction with other transcription factors. PMID- 3029567 TI - Cloning of a human S-phase cell cycle gene: use of transient expression for screening. AB - We report here the cloning of a human cell cycle gene capable of complementing a temperature-sensitive (ts) S-phase cell cycle mutation in a Chinese hamster cell line. Cloning was performed as follows. A human genomic library in phage lambda containing 600,000 phages was screened with labeled cDNA synthesized from an mRNA fraction enriched for the specific cell cycle gene message. Plaques containing DNA inserts which hybridized to the cDNA were picked, and their DNAs were assayed for transient complementation in DNA transformation experiments. The transient complementation assay we developed is suitable for most cell cycle genes and indeed for many genes whose products are required for cell proliferation. Of 845 phages screened, 1 contained an insert active in transient complementation of the ts cell cycle mutation. Introduction of this phage into the ts cell cycle mutant also gave rise to stable transformants which grew normally at the restrictive temperature for the ts mutant cells. PMID- 3029566 TI - Factors influencing alternative splice site utilization in vivo. AB - To study factors that influence the choice of alternative pre-mRNA splicing pathways, we introduced plasmids expressing either wild-type or mutated simian virus 40 (SV40) early regions into tissue culture cells and then measured the quantities of small-t and large-T RNAs produced. One important element controlling splice site selection was found to be the size of the intron removed in the production of small-t mRNA; expansion of this intron (from 66 to 77 or more nucleotides) resulted in a substantial increase in the amount of small-t mRNA produced relative to large-T mRNA. This suggests that in the normal course of SV40 early pre-mRNA processing, large-T splicing is at a competitive advantage relative to small-t splicing because of the small size of the latter intron. Several additional features of the pre-mRNA that can influence splice site selection were also identified by analyzing the effects of mutations containing splice site duplications. These include the strengths of competing 5' splice sites and the relative positions of splice sites in the pre-mRNA. Finally, we showed that the ratio of small-t to large-T mRNA was 10 to 15-fold greater in human 293 cells than in HeLa cells or other mammalian cell types. These results suggest the existence of cell-specific trans-acting factors that can dramatically alter the pattern of splice site selection in a pre-mRNA. PMID- 3029565 TI - Molecular characterization of novel reciprocal translocation t(6;14) in an Epstein-Barr virus-transformed B cell precursor. AB - An in vitro culture of FLEB14 cells, an Epstein-Barr virus-transformed B cell precursor containing the germ line immunoglobulin genes, gave rise to a uniclonally expanded variant, FLEB14 delta 3, which was rearranged at the immunoglobulin heavy-chain gene locus. Cytogenetic analysis showed that FLEB14 delta 3 had a novel reciprocal translocation, t(6;14)(q15;q32). Molecular cloning of the rearranged DNA fragments and determination of their nucleotide sequence revealed that the recombination event was reciprocal, imprecise, and nonhomologous and took place in the S mu region, like those found in Burkitt's lymphoma cells. We propose a molecular model to explain this genetic event which may be relevant to class switch recombination. The translocated sequence of chromosome 6 did not contain any known oncogenes, although the sequence is conserved among mammals. FLEB14 delta 3 did not show tumorigenicity. PMID- 3029568 TI - At least two nuclear proteins bind specifically to the Rous sarcoma virus long terminal repeat enhancer. AB - We used the sensitive gel electrophoresis DNA-binding assay and DNase I footprinting to detect at least two protein factors (EFI and EFII) that bound specifically to the Rous sarcoma virus (RSV) enhancer in vitro. These factors were differentially extracted from quail cell nuclei, recognized different nucleotide sequences in the U3 region of the RSV long terminal repeat, and appeared to bind preferentially to opposite DNA strands as monitored by the DNase I protection assay. The EFI- and EFII-protected regions within U3 corresponded closely to sequences previously demonstrated by deletion mutagenesis to be required for enhancer activity, strongly suggesting a functional significance for these proteins. Only weak homologies between other enhancer consensus sequence motifs and the EFI and EFII recognition sites were observed, and other viral enhancers from simian virus 40 and Moloney murine sarcoma virus did not compete effectively with the RSV enhancer for binding either factor. PMID- 3029569 TI - Multiple domains for the chicken cellular sequences homologous to the v-ets oncogene of the E26 retrovirus. AB - We have investigated the structure of chicken genomic DNA homologous to v-ets, the second cell-derived oncogene of avian retrovirus E26. We isolated a c-ets locus spanning ca. 30.0 kilobase pairs (kbp) in the chicken genome with homologies to 1,202 nucleotides (nt) of v-ets (total length, 1,508 nt) distributed in six clusters along 18.0 kbp of the cloned DNA. The 5'-distal part of v-ets (224 nt) was homologous to chicken cellular sequences contained upstream within a single 16.0-kbp EcoRI fragment as two typical exons but not found transcribed into the major 7.5-kb c-ets (or 4.0-kb c-myb) RNA species. Between these two v-ets-related cellular sequences we found ca 40.0 kbp of v-ets unrelated DNA. Finally, the most 3' region of homology to v-ets in the cloned DNA was shown to consist of a truncated exon lacking the nucleotides coding for the 16 carboxy-terminal amino acids of the viral protein but colinear to one of the two human c-ets loci, c-ets-2. PMID- 3029570 TI - Regulated expression of a complete human beta-globin gene encoded by a transmissible retrovirus vector. AB - We introduced a human beta-globin gene into murine erythroleukemia (MEL) cells by infection with recombinant retroviruses containing the complete genomic globin sequence. The beta-globin gene was correctly regulated during differentiation, steady-state mRNA levels being induced 5- to 30-fold after treatment of the cells with the chemical inducer dimethyl sulfoxide. Studies using vectors which yield integrated proviruses lacking transcriptional enhancer sequences indicated that neither retroviral transcription nor the retroviral enhancer sequences themselves had any obvious effect on expression of the globin gene. Viral RNA expression also appeared inducible, being considerably depressed in uninduced MEL cells but approaching normal wild-type levels after dimethyl sulfoxide treatment. We provide data which suggest that the control point for both repression and subsequent activation of virus expression in MEL cells lies in the viral enhancer element. PMID- 3029572 TI - The receptor for immunoglobulin E: examination for kinase activity and as a substrate for kinases. AB - Purified receptor-immunoglobulin E (IgE) complexes incubated with [gamma-32P]-ATP incorporated phosphorus into tyrosines on the beta and gamma chains of the receptor. The activity had the typical characteristics of a tyrosine kinase. Immunoprecipitation of the complexes with anti-IgE left the activity in the supernatant, demonstrating that the receptor itself was not the kinase. The receptor was also phosphorylated when membranes or intact cells were incubated with radioactive ATP or phosphate, respectively, but in each case the subunits or amino acid residues that were modified were distinctive. PMID- 3029571 TI - The excised leader of human cytochrome c oxidase subunit I mRNA which contains the origin of mitochondrial DNA light-strand synthesis accumulates in mitochondria and is polyadenylated. AB - We identified a polyadenylated RNA species which contains the origin of human mitochondrial DNA light-strand synthesis and the surrounding complementary sequences of the four light-strand-encoded tRNAs. This RNA (RNA 9L) is probably derived from the leader portion of RNA 6 which is excised during the formation of the mature cytochrome c oxidase subunit mRNA (RNA 9). The high degree of secondary structure of this RNA is presumably responsible for its anomalous electrophoretic behavior in denaturing polyacrylamide gels. PMID- 3029574 TI - [Pathology of fetal cystic fibrosis: morphological changes]. PMID- 3029573 TI - Regulation of complement proteins C2 and factor B by interleukin-1 and interferon gamma acting on transfected L cells. PMID- 3029575 TI - [Granular cell tumor (Abrikosov myoblastoma) of the respiratory tract associated with epithelial hyperplasia simulating carcinoma]. PMID- 3029577 TI - [Genetic characteristics and physical organization of the R-plasmid pBS52 with a broad range of bacterial hosts]. AB - The genetic and physical data on Pseudomonas aeruginosa plasmid pBS52 coding for the resistances to ampicillin, streptomycin and sulfonamids have been obtained. This conjugative plasmid is transferable to a broad range of gram-negative bacterial hosts and compatible with the broad host-range plasmids from all known incompatibility groups. The plasmid size has been determined (38 Kb) and a physical map has been constructed using restriction endonucleases EcoRI, EcoRV, BamHI, BglII, PstI, PvuII, SalI, SlaI. The presence of a fragment, approximately 200 bp in size, which contains the sites for many of widely used restriction endonucleases is a characteristic feature of the plasmid pBS52. PMID- 3029576 TI - [Irreversible changes in phage DNA after its protonation in solution and inside the virion detected by the transfection method]. AB - The dependence of irreversible structural changes in phage lambda DNA on the degree of its protonation in a solution and inside the virion has been found by measuring the transfection activity of bacteriophage. The different effect of ionic strength on pH-dependence of the irreversible changes in the structure of DNA upon its protonation in a solution or in situ has been registered and explained. The insignificant shift of pH from neutral region value in 0.1 M NaCl has resulted in a damaging effect of H+ ions on compact DNA in situ as compared to the DNA in a solution. The effect of H+ ions on compact DNA in situ is mainly based on the formation of noncovalent intermolecular DNA-protein and DNA-DNA linkages. PMID- 3029578 TI - [Purification and properties of 2 restriction endonucleases from Bacillus subtilis]. PMID- 3029579 TI - [Comparative restriction analysis of HindIII-F-fragments of DNA from 4 strains of vaccinia virus]. AB - The localization of KpnI, SacI, XhoI, AvaI, PstI, BglI, BamHI, EcoRI, PmiI, SalI, BglII, restriction endonuclease cleavage sites in HindIII-F-fragments of DNA from vaccinia strains WR, Copenhagen, LIVP and neurovaccine has been detected. The fragments have been shown to differ in the number of AvaI, EcoRI and BamHI sites. The fragments also differ from the analogue of Tian Tan vaccinia strain in the pattern of restriction by AvaI, XhoI, PstI, EcoRI and BamHI endonucleases. PMID- 3029580 TI - [Transposon-mediated integration of the RP1 plasmid into chromosomes of methylotrophic bacteria]. AB - The homology region between the DNA of plasmid RP1ts::Tn601 and chromosome of the thermotolerant methylotrophic bacterium Methylobacterium sp. SKF240 has been constructed by transposon Tn601 translocation into the chromosome. The clones of Methylobacterium sp. SKF240 having integrated the plasmid RP1 into the chromosome have been obtained by conjugation on the basis of above mentioned genetic technique. The integration of plasmid RP1 into the chromosomal DNA of the methylotroph has been confirmed by the genetic and electrophoretic methods. Clones harbouring the integrated plasmid are able to transfer the chromosomal genes for methionine and isoleucine-valine synthesis to the recipient cells of P. aeruginosa PAO ML4262 by conjugation. PMID- 3029581 TI - Induction of the SOS system in a dam-3 mutant: a diagnostic strain for chemicals causing DNA mismatches. AB - We have developed a strain of E. coli in which expression of the SOS function sfiA, monitored by means of a sfiA::lacZ operon fusion, is efficiently triggered by the two base analogues 2-aminopurine and 5-bromo-2'-deoxyuridine. This strain resulted from introduction of a dam-3 mutation into a Uvr+, Rfa+ derivative of strain PQ37 used in the SOS chromotest, a bacterial colorimetric assay for genotoxins (Quillardet et al., 1982). The dam-3 mutation affects the mismatch correction system in E. coli. We show that the SOS-inducing capacity of a weak SOS inducer such as the alkylating agent ethyl methanesulfonate was also increased in the dam-3 strain. We provide evidence that the increase in SOS inducibility due to the dam-3 mutation is specific for compounds causing DNA mismatches and propose the use of the dam-3 derivative of PQ37 as a diagnostic strain for such agents. This diagnostic strain can be a useful addition to the SOS chromotest. PMID- 3029582 TI - Effect of irradiation and mutagenic chemicals on the generation of ADH2 constitutive mutants in yeast. Significance for the inducibility of Ty transposition. AB - Mutations caused by the insertion of a Ty element resulting in an antimycin-A resistant phenotype in an adh1- strain of Saccharomyces cerevisiae were used as an assay for the quantitative detection of Ty transposition. Antimycin-A resistant mutants were found to be inducible by ethyl methanesulfonate (EMS) as well as by gamma- and UV irradiation. DNA analysis of gamma-induced mutants showed an increase of the fraction of Ty insertions in the ADH2 locus with increasing dose. PMID- 3029583 TI - Intranuclear localization of UV-induced DNA repair in human VA13 cells. AB - We have investigated the intranuclear localization of DNA-repair synthesis in G1 phase VA13 human cells. Ultraviolet-irradiated cells were permitted to perform unscheduled DNA synthesis in 3H-thymidine (3H-TdR) and then extracted with nonionic detergent and 2 M NaCl to produce nucleoids in which residual nuclear matrix was surrounded by an extended halo of DNA loops. Autoradiographic analysis of these structures permitted discrimination of DNA repair between the matrix and halo regions. Repair label in nucleoids prepared from cells after exposure to fluences of 2.5-30 J/m2 exhibited a dose-dependent association with the nuclear matrix, which ranged from 80% after 2.5 J/m2 to 50% after 30 J/m2. These results support the view that DNA repair is a nuclear matrix-associated process. This conclusion is in agreement with our preliminary study (Harless et al., 1983) and the results of McCready and Cook (1984) but contrasts with that of Mullenders et al. (1983). Questions concerning the differing experimental designs and their potential effects on the localization of DNA repair are discussed. The implications of these results to previous attempts to isolate chromatin factors associated with DNA repair are also considered. PMID- 3029584 TI - An immortalized xeroderma pigmentosum, group C, cell line which replicates SV40 shuttle vectors. AB - We have established and characterized an immortalized xeroderma pigmentosum (XP), group C, cell line. Transformation of the human fibroblasts was carried out with a recombinant plasmid, pLAS-wt, containing SV40 DNA encompassing the entire early region with a defective origin of DNA replication. The transformed XP cell line, XP4PA-SVwt, and the normal transformed fibroblasts AS3-SVwt, both express SV40 T antigen together with enhanced levels of the transformation-associated cellular protein, p53. XP4PA-SVwt retains the XP UV-repair defective phenotype as demonstrated by low levels of unscheduled DNA synthesis and by the reduced survival of irradiated SV40 virus. Analysis of cellular DNA shows a single major, stable, integration site of pLAS-wt in the XP4PA-SVwt cells. The T antigen in these cells supports efficiently the replication of SV40 based shuttle vectors and should prove suitable for the introduction, expression and selection of genes related to DNA repair and to the study of mutagenesis using defined molecular probes. PMID- 3029585 TI - Influence of post X-irradiation treatment with Neurospora endonuclease on the symmetry of chromatid interchanges in CHO cells in vitro. PMID- 3029586 TI - Inducible DNA polymerase I synthesis in a UV hyper-resistant mutant of Escherichia coli. AB - A mutant of Escherichia coli which is more resistant to shortwave UV light than its wild-type parent strain and which can synthesise DNA polymerase I constitutively has been further analysed. It carries two mutational alleles which are located about 1.5 min apart and cotransducible by P1 with the argH locus. The two mutational alleles have been segregated and their analysis shows that one of them is responsible for UV hyper-resistance whereas the other mutation confers UV sensitivity. Recombinant plasmids carrying various sections of the polA regulatory region, linked to a galK gene, were introduced into the mutant strains. Analysis of galactokinase shows that the enzyme activity in the UV hyper resistant mutant is increased. The results suggest that the synthesis of DNA polymerase I in E. coli is inducible. PMID- 3029587 TI - Acute arsenic intoxication presenting as Guillain-Barre-like syndrome. AB - Arsenic-induced polyneuropathy is traditionally classified as an axonal-loss type, electrodiagnostically resulting in low amplitude or absent sensory and motor responses, relatively preserved proximal and distal motor conduction rates, and distal denervation. We report four patients with a subacute onset progressive polyradiculoneuropathy following high-dose arsenic poisoning. In three patients, early electrodiagnostic testing demonstrated findings suggestive of an acquired segmental demyelinating polyradiculoneuropathy. Serial testing confirmed evolution into features of a distal dying-back neuropathy. We hypothesize that arsenic toxicity and the resultant biochemical derangement of the peripheral nerve cell leads to subtle changes in axonal function that produce, initially, segmental demyelination and eventually distal axonal degeneration. Acute arsenic toxicity must be suspected in patients with clinical and electrodiagnostic features supporting Guillain-Barre syndrome. PMID- 3029588 TI - Neurotoxic effects of organophosphorus insecticides. An intermediate syndrome. AB - Acute neurotoxic effects during the cholinergic phase of organophosphorus insecticide poisoning and delayed neurotoxic effects appearing two to three weeks later are well recognized. We observed 10 patients who had paralysis of proximal limb muscles, neck flexors, motor cranial nerves, and respiratory muscles 24 to 96 hours after poisoning, after a well-defined cholinergic phase. The compounds involved were fenthion, monocrotophos, dimethoate, and methamidophos. Four patients urgently required ventilatory support. The paralytic symptoms lasted up to 18 days. A delayed polyneuropathy later developed in one patient. Three patients died. Electromyographic studies showed fade on tetanic stimulation, absence of fade on low-frequency stimulation, and absence of post-tetanic facilitation, suggestive of a postsynaptic defect. This neuromuscular junctional defect may have been the predominant cause of the paralytic symptoms, with neural and central components contributing to various degrees. Our patients appeared to have a distinct clinical entity (a so-called intermediate syndrome) that developed after the acute cholinergic crisis and before the expected onset of the delayed neuropathy. PMID- 3029589 TI - Changing profile of pesticide poisoning. PMID- 3029590 TI - Human infection with Ehrlichia canis, a leukocytic rickettsia. PMID- 3029591 TI - Inhibition of angiotensin-converting enzyme in congestive heart failure. PMID- 3029592 TI - Chemotherapy with or without radiation therapy in limited small-cell carcinoma of the lung. AB - We conducted a prospective, randomized study to clarify the role of radiotherapy of the primary tumor in limited small-cell cancer of the lung. After stratification for sex and for performance score based on the ability to ambulate, patients were randomly assigned to receive initial radiotherapy plus chemotherapy, delayed radiotherapy plus chemotherapy, or chemotherapy alone. The chemotherapy consisted of cyclophosphamide, etoposide (VP-16-213), and vincristine, with doxorubicin subsequently replacing etoposide in alternate cycles 7 through 18. Chemotherapy was given every three weeks for 18 months. The radiotherapy comprised 4000 rad in four weeks, followed by a 1000-rad "boost" directed against residual disease. All patients received prophylactic whole-brain radiation. The patients enrolled totaled 426, and 399 were evaluable. There was a statistically significant difference in the frequency of complete responses in favor of the two radiotherapy regimens (P = 0.0013). Failure-free survival was also longer with these two regimens (P less than 0.001), as was the interval before treatment failure in the chest (P less than 0.001) and overall survival (P = 0.0099). As expected, toxic effects--chiefly neutropenia--were also increased. The addition of radiotherapy of the primary tumor to combination chemotherapy improved both complete-response rates and survival, with increased but acceptable toxicity. PMID- 3029593 TI - Independent phosphatidylinositol synthesis in pituitary plasma membrane and endoplasmic reticulum. AB - Phosphatidylinositol (PtdIns), the most abundant phosphoinositide, is the precursor of phosphatidylinositol 4-monophosphate which is converted to phosphatidylinositol 4,5-bisphosphate, the lipid hydrolysed as an early step in signal transduction by many stimuli. It is generally thought that a single enzyme in the endoplasmic reticulum, PtdIns synthase (CDP-diglyceride:myoinositol 3 phosphatidyltransferase, EC 2.7.8.11), is responsible for PtdIns synthesis and that newly synthesized PtdIns is transported to the plasma membrane by exchange proteins. Several investigators have proposed that there are two functionally distinct pools of PtdIns, one responsive to stimulation and the other not, and that the stimulus-responsive pool may be synthesized at a different site within the cell, perhaps within the plasma membrane. Indeed, it was suggested that there is PtdIns synthase activity in plasma membrane isolated from rat liver. GH3 rat pituitary tumour cells are an excellent model system to study stimulation of phosphoinositide metabolism by thyrotropin-releasing hormone (TRH). Conversion of PtdIns to polyphosphoinositides and TRH (and GTP)-activated phosphoinositide hydrolysis are known to occur in plasma membrane isolated from GH3 cells. Here we report that PtdIns synthase activity in the plasma membrane of GH3 cells is distinct from that present in the endoplasmic reticulum. The plasma membrane PtdIns synthase may be responsible for a portion of PtdIns re-synthesis that occurs during cell stimulation. PMID- 3029594 TI - Homology between Streptomyces genes coding for synthesis of different polyketides used to clone antibiotic biosynthetic genes. AB - Many important antibiotics such as tetracyclines, erythromycin, adriamycin, monensin, rifamycin and avermectins are polyketides. In their biosynthesis, multifunctional synthases catalyse iterated condensation of thio-esters derived from acetate, propionate or butyrate to yield aliphatic chains of varying length and carrying different alkyl substituents. Subsequent modifications, including aromatic or macrolide ring closure or specific methylations or glycosylations, generate further chemical diversity. It has been suggested that, if different polyketide synthases had a common evolutionary origin, cloned DNA coding for one synthase might be used as a hybridization probe for the isolation of others. We show here that this is indeed possible. Study of a range of such synthase genes and their products should help to elucidate what determines the choice and order of condensation of different residues in polyketide assembly, and might yield, by in vitro recombination or mutagenesis, synthase genes capable of producing novel antibiotics. Moreover, because genes for entire antibiotic pathways are usually clustered in Streptomyces, cloned polyketide synthase genes are valuable in giving access to groups of linked biosynthetic genes. PMID- 3029595 TI - Protein--DNA interaction. Another folding problem? PMID- 3029596 TI - AIDS meeting calls for wider testing. PMID- 3029597 TI - Retroviral transduction of T-cell antigen receptor beta-chain and myc genes. AB - Support for multistage models of oncogenesis has been provided by several highly leukaemogenic retrovirus isolates that have transduced more than one host cell gene. Where functional studies have been performed, these retroviral oncogenes show synergy for in vitro transformation and leukaemogenesis. In naturally occurring feline leukaemias associated with feline leukaemia virus (FeLV), retroviral transduction of myc is a frequent oncogenic mechanism. But evidence suggesting that the FeLV v-myc genes might be insufficient for leukaemogenesis was provided by the latency (12 weeks) and clonality of FeLV/v-myc-induced tumours and the absence of demonstrable in vitro transformation by these viruses. In the search for secondary leukaemogenic events in FeLV/v-myc tumours, we have identified a case of FeLV transduction of a T-cell antigen receptor beta-chain gene. The proviruses carrying this gene (which we have named v-tcr) were a separate population from those carrying v-myc. In its normal role, the T-cell receptor beta-chain forms part of a multimeric complex involved in antigen recognition and T-cell activation. We suggest that v-tcr is a novel viral oncogene which assisted v-myc in the genesis of a naturally occurring case of thymic lymphosarcoma. PMID- 3029598 TI - Cell fusion in viral diseases. PMID- 3029599 TI - A potential animal model for Lesch-Nyhan syndrome through introduction of HPRT mutations into mice. AB - The human Lesch-Nyhan syndrome is a rare neurological and behavioural disorder, affecting only males, which is caused by an inherited deficiency in the level of activity of the purine salvage enzyme hypoxanthine-guanosine phosphoribosyl transferase (HPRT). How the resulting alterations in purine metabolism lead to the severe symptoms characteristic of Lesch-Nyhan patients is still not understood. No mutations at the Hprt locus leading to loss of activity have been described in laboratory animals. To derive an animal model for the Lesch-Nyhan syndrome, we have used cultured mouse embryonic stem cells, mutagenized by retroviral insertion and selected for loss of HPRT activity, to construct chimaeric mice. Two clonal lines carrying different mutant Hprt alleles have given rise to germ cells in chimaeras, allowing the derivation of strains of mutant mice having the same biochemical defect as Lesch-Nyhan patients. Male mice carrying the mutant alleles are viable and analysis of their cells shows a total lack of HPRT activity. PMID- 3029600 TI - Ascaris DNA topoisomerase I binds preferentially to the germ-line- limited DNA. PMID- 3029601 TI - Antibodies labeled with metallic radionuclides: influence of nuclide chemistry on dose distribution. AB - An antibody with human CEA specificity has been labeled with either yttrium-90, scandium-47, or indium-111, via a diethylenetriamine pentaacetic acid (DTPA) link covalently bound to the protein. The clearance of these proteins from the blood of mice can be described by a single exponential; the half-life decreases in the order indium-111 greater than yttrium-90 greater than scandium-47. Associated with the blood clearance is an uptake of radioactivity into the liver; scandium 47 has the highest concentration, indium-111 has the least, and yttrium-90 is intermediate. There is no correlation between these results and the equilibrium stability constants of the metals with DTPA-like ligands. The results obtained show that, in vivo, scandium-47 and yttrium-90 are more easily displaced from DTPA by other ions than is indium-111. They also show that free DTPA is able to extract yttrium-90 and scandium-47, but not indium-111, from the liver of treated animals, indicating that indium-111 is resistant to ligand exchange reactions in vivo. These data indicate that 1) the equilibrium stability constant is not a good indicator of the in vivo stability of metal-labeled proteins and 2) it is possible to manipulate the ion distribution and therefore the dose from scandium 47 and yttrium-90 after injection of the labeled proteins. PMID- 3029602 TI - Radioimmunoimaging of human small cell lung carcinoma xenografts in nude mice receiving several monoclonal antibodies. AB - Small cell carcinoma of the lung is a highly lethal disease, killing nearly all patients afflicted with it. Although it metastasizes early and widely, it is exquisitely radiosensitive, and transient responses to this form of therapy are often dramatic. We have recently developed several monoclonal antibodies (MAb) reactive with antigens associated with this tumor. Scintigraphic and dual-label evaluations of their localization in nude mice bearing small cell carcinoma xenografts (derived from the human small cell carcinoma cell line NCI H69) were undertaken as a preliminary step in the eventual evaluation of their therapeutic potential. Anti-small cell MAb UM-MS1, 2, and 3, were labeled with 131I with the Iodobead method. Labeling efficiencies ranged from 68% to 88%. A control MAb, 225.28S (antimelanoma), was labeled with 125I by the same method. Although the labeled anti-small cell antibodies bound 20-40 times more to NCI H69 cells than did the antimelanoma control antibody, their total immunoreactive fraction was relatively low. Separate groups of nude mice bearing NCI H69-derived tumors were studied for imaging and localization with each 131I-labeled MAb. Successful imaging was achieved with two of the MAb; the quality of the images ranged from moderate to excellent. However, much of the localization to the tumors was due to nonspecific factors, as evidenced by the considerable tumor accretion of the control antibody.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3029603 TI - Low-dose-rate radiation sensitivity of a human hepatoma cell line. AB - A human hepatoma cell line, HepG2, is being used in a study of the radiobiologic factors that determine the effectiveness of radiolabeled immunoglobulin therapy of primary hepatoma. Growth inhibition by continuous low-dose-rate radiation occurs at lower dose rates for HepG2 than for most other cell lines studied. Based on survival curves following acute radiation, HepG2 appears to have less sublethal damage repair than most human tumors. Prior treatment with acute radiation did not sensitize HepG2 to either growth inhibition or cell killing by low-dose-rate radiation. PMID- 3029604 TI - Solubilization and characterization of the 3H-desipramine binding site of rat phaeochromocytoma cells (PC12-cells). AB - 3H-Desipramine binding sites of the plasma membranes of rat phaeochromocytoma cells (PC12-cells) were solubilized with the nonionic detergent digitonin (0.5%). With the method described here, the binding characteristics of the desipramine binding site were essentially unaltered by solubilization. Binding of 3H desipramine to the solubilized binding site showed the following characteristics: (1) 3H-desipramine bound with high affinity (KD = 16.6 nmol/l) to a single class of noninteracting (Hill-coefficient = 1.01) binding sites; (2) binding was reversible; (3) binding of unlabelled desipramine had the same dissociation constant as had 3H-desipramine; (4) increasing concentrations of sodium- and chloride-ions stimulated the binding of 3H-desipramine; (5) binding was inhibited by various inhibitors and substrates of neuronal uptake of noradrenaline; and (6) inhibition of binding by the optical isomers of cocaine, oxaprotiline, and amphetamine showed marked stereoselectivity (with preference for (-)cocaine, (+)oxaprotiline, and (+)amphetamine). The finding that the binding of 3H desipramine to the solubilized binding site was dependent on sodium and chloride, as the neuronal uptake of noradrenaline is, and the finding that all substrates of uptake inhibited the binding of 3H-desipramine, is consistant with the view that desipramine binds to the substrate recognition site of the neuronal carrier for noradrenaline. PMID- 3029605 TI - Sympathetic denervation and alpha adrenoceptors in dog cerebral arteries. AB - The magnitude of the noradrenaline-induced contractions of dog middle cerebral arteries was less than that seen in the vertebral, common carotid, femoral and renal arteries. Noradrenaline and clonidine produced a similar magnitude of maximum contractions in the middle cerebral arteries, whereas methoxamine produced no significant contractions in the same arteries. In the extracranial arteries, noradrenaline and methoxamine produced significantly larger contractions than clonidine. Binding studies revealed no specific 3H-prazosin binding sites in the cerebral arteries, though such binding sites were evident in the case of extracranial arteries. 3H-Yohimbine binding studies revealed the presence of two classes of binding sites with high and low affinities in both cerebral and extracranial arteries. After superior cervical ganglionectomy, noradrenaline- and clonidine-induced contractions of the denervated middle cerebral arteries were not altered, compared with the control arteries. A 3H yohimbine binding study was also performed using the denervated cerebral arteries. This study revealed that there was a low affinity 3H-yohimbine binding site, whereas high affinity 3H-yohimbine binding site was not detectable. These results suggest the presence of two different binding sites with high and low affinity for alpha 2 adrenoceptors, which we are classifying into alpha 2H and alpha 2L subtypes. The high affinity sites, alpha 2H adrenoceptors, are presynaptically located while the low affinity sites, alpha 2L adrenoceptors, located postsynaptically. The noradrenaline-induced contractions are probably mediated by postsynaptic low affinity sites of alpha 2 adrenoceptors (alpha 2L adrenoceptors) in the cerebral arteries and mainly by alpha 1 adrenoceptors in the extracranial arteries. PMID- 3029606 TI - Influence of dietary fiber and intraluminal pressure on absorption and pre epithelial diffusion resistance (unstirred layer) in rat jejunum in situ. AB - The appearance rates of antipyrine, benzoic acid, benzylamine, urea, and alpha methyl-D-glucoside (MG) in jejunal venous blood of anesthetized rats were measured with and without dietary fibers methylcellulose, carboxymethylcellulose sodium, guaran, and sodium alginate in the luminal solution. Raising the concentration of methylcellulose from 0 to 17.5 g/l resulted in an exponential increase in the viscosity of the solution to 98 cSt, a linear decrease of the diffusion coefficient for antipyrine by 28%, and an increase in antipyrine absorption in the perfused jejunal segment by 23%. The simultaneous increase in intraluminal pressure and radius resulted in a linear relation between absorption rate and apparent mucosal surface area. Similar results were obtained by raising intraluminal pressure directly using a carbohydrate-free perfusion solution. In the perfused rat jejunum, the effect of increased pre-epithelial diffusion resistance (i.e. reduced diffusion coefficient and lengthened diffusion distance) induced by methylcellulose on absorption was overcome by the effect of the enlarged apparent mucosal surface area. Preperfusion of a substrate-free, guaran containing solution followed by perfusion with a guaran-free solution containing antipyrine and MG retarded the increase in the appearance rate of these substrates due to the additional viscous guaran layer left after preperfusion. Constant distension of the intestinal wall was achieved by injecting 0.5 ml of the solution into a closed jejunal segment. Addition of the carbohydrates to the injection solution (approx. 100 cSt viscosity) resulted in a 3% to 20% reduction in the diffusion coefficients and in the absorption of antipyrine, benzoic acid, and MG. Diffusion coefficients for urea and benzylamine were reduced by 5% to 12%; absorption varied in the range of the control (-22% to +43%). Model analysis revealed that, in the closed jejunal segment of the rat, the limiting step in the absorption process of antipyrine, benzoic acid, and MG was pre-epithelial diffusion resistance; the reduction of absorption, therefore, corresponded roughly to that of the diffusion coefficient. In the case of urea and benzylamine, pre-epithelial diffusion resistance was only 20% of the total permeation resistance: the influence of the polymers on absorption, therefore, was not always significant. PMID- 3029608 TI - The pseudo-false positive Meckel's scan. PMID- 3029607 TI - Interaction of choline with muscarine receptor-stimulated phosphoinositide metabolism in the rat brain. AB - Previous receptor binding studies had shown that choline can interact with low potency with muscarine cholinoceptors. In the present study we have investigated whether choline is capable of functionally activating muscarine receptors by investigating its ability in stimulating the hydrolysis of phosphoinositides, a response believed to be coupled in brain to the M1 subtype of muscarine receptors. The results indicated that choline was only a very weak inducer of inositol phosphates (InsPs) accumulation in rat cerebral cortex slices as compared with acetylcholine or charbachol. Maximal increase of InsPs accumulation, at a choline concentration of 10 mM, was only 39 +/- 7%, as compared with the 4- to 6-fold stimulation induced by the other compounds. This effect of choline was not modified by physostigmine nor by the uptake inhibitor hemicholinium-3. At high concentrations, however, choline antagonized the stimulatory effect of acetylcholine and carbachol, suggesting that it might act as a partial agonist at this subtype of muscarine receptors, similar to what has been observed with oxotremorine. Choline had no effect on noradrenaline stimulated InsPs accumulation. PMID- 3029610 TI - [Microangiopathic hemolytic anemia and metastasized carcinoma]. PMID- 3029609 TI - [A patient with industrial hydrocyanic acid poisoning]. PMID- 3029611 TI - Paget's disease of the male breast. PMID- 3029612 TI - [Renal transport of uric acid: influence of treatment by converting enzyme inhibitors]. PMID- 3029613 TI - Effects of chronic intracerebroventricular GABAergic treatment on basal and chronic or acute stress-induced adrenocortical activities in the thalamic pigeon. AB - Muscimol was chronically administrated to the third ventricle of thalamic pigeons by means of osmotic minipumps at the rate of 0.25 microgram X h-1 for 28 days. No abnormal behavioural sign was noted. The animals were subjected daily to chronic intermittent stress for the same 28-day period. Basal and stress-induced adrenocortical activities were evaluated by recording serial plasma corticosterone levels at the end of the experimental session. Untreated controls exhibited both components of the adaptation of the adrenocortical response to chronic stress: attenuation, i.e., a decrease in magnitude and disappearance of the late rebounding phenomenon, and anticipation, i.e., the occurrence of a conditioned component before stress itself. The adaptation to chronic stress was partly impaired by GABAergic treatment. The anticipatory conditioned peak subsisted but the magnitude of the post-stress peak was found not to be reduced whereas rebounding events were suppressed after chronic as well as acute stress. The basal resting levels of corticosterone were significantly lowered in muscimol treated animals. A lesion placement in the anterior dorsomedial thalamus (ADMT) resulted in the same profile of stress-induced plasma corticosterone levels as seen after muscimol administration. Adaptation did not develop in ADMT animals and GABAergic stimulation, either acute or chronic, had no effect on their response to stress. PMID- 3029614 TI - Estradiol modulates density of putative 'oxytocin receptors' in discrete rat brain regions. AB - Previous studies have provided evidence for a discrete localization of two types of vasopressin (AVP)-labeled binding sites in the rat brain, i.e., regions labeled preferentially with AVP (putative AVP receptors) and regions labeled with AVP as well as oxytocin (OT). The latter binding sites are considered here as putative OT receptors. In the present study the effect of estradiol on the number of these putative receptor sites for OT and AVP was investigated in rat brain after daily subcutaneous administration of the steroid (10 micrograms/100 g body weight) to ovariectomized rats. Specific binding of [3H]-OT and [3H]-AVP was determined after in vitro incubation of frozen brain sections, autoradiography and quantitation of the images with computer-assisted densitometry. Estradiol increased the number of OT receptors at least 4-fold in the ventromedial nucleus of the hypothalamus, regions of the olfactory tubercle, the nucleus accumbens and occasionally in the organum vasculosum laminae terminalis. A smaller increase (two-fold) was noted in the central amygdala, while a tendency to a decrease in OT receptor number was noted in the olfactory nucleus and the ventral subiculum. Estradiol treatment permitted an estimation of binding constants of [3H]-OT binding to a membrane fraction of microdissected ventromedial hypothalamic region (Kd: 1.3 nM, Bmax: 19.9 fmol/mg protein). The number of putative AVP receptors in the lateral septum and in the nucleus tractus solitarii was not affected by estradiol. In conclusion, the OT receptor system is subject to modulation by estradiol in some discrete brain regions, but not in others.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3029615 TI - Dopaminergic regulation of the estrogen-induced glandular kallikrein in the rat anterior pituitary. AB - Glandular kallikrein is a major estrogen-induced protein of the rat anterior pituitary. A second kallikrein-like protease in the rat anterior pituitary (kallikrein A) is not affected by estrogens, nor is a third pituitary protease which cleaves a trypsin substrate but not kallikrein substrates. This study examined whether any of the pituitary proteases are regulated by dopaminergic mechanisms. Ovariectomized female rats were treated for 5-10 days with reserpine (a catecholamine depleting agent), haloperidol (a dopamine receptor blocker) or bromocriptine (a dopamine receptor agonist); some rats also received 1 or 2 micrograms estradiol benzoate every 48 h. Following activation of latent proteases with trypsin, anterior pituitary extracts were assayed for kallikrein activity before and after fractionation on DEAE-Sephadex to separate the two kallikrein-like proteases. Reserpine or haloperidol doubled glandular kallikrein levels in anterior pituitaries from estrogen-treated rats. Reserpine or haloperidol had little or no effect in the absence of estrogen (estrogen produced a 5- to 7-fold increase in glandular kallikrein in the absence of drug treatment). Bromocriptine markedly attenuated the ability of estrogen to induce glandular kallikrein. Further, bromocriptine blocked the ability of reserpine to increase glandular kallikrein levels, and haloperidol attenuated the effect of bromocriptine. Other anterior pituitary proteases were unaffected by either estrogen, haloperidol, reserpine or bromocriptine. The results demonstrate that the estrogen induction of glandular kallikrein in the rat anterior pituitary is modulated by inhibitory dopaminergic mechanisms. Prolactin is the only pituitary hormone which exhibits a similar profile of hormonal and neuroendocrine regulation; this suggests a possible link between glandular kallikrein and prolactin. PMID- 3029616 TI - Elevated plasma levels of pro-opiomelanocortin-derived peptides in sheep following hypothalamo-pituitary disconnection. AB - We have studied the control of the pituitary-adrenal axis in ovariectomized sheep following hypothalamo-pituitary disconnection (HPD) by following plasma levels of immunoreactive (ir-) products of pro-opiomelanocortin (POMC). In ovariectomized HPD ewes, gonadotropin levels were below detection limits. In contrast, levels of ir-ACTH were modestly but significantly elevated over those in matched ovariectomized control ewes, though cortisol levels were not significantly different; levels of ir-alpha MSH and ir-beta EP (beta-endorphin) were substantially and significantly raised in HPD compared with control plasma. Size exclusion HPLC showed plasma beta EP/beta LPH (beta-lipotropin) levels to be higher in HPD than control ewes. Dexamethasone administration lowered plasma ir ACTH but not ir-beta EP; in contrast, bromocriptine lowered ir-beta EP but not ir ACTH. We interpret these data as evidence (1) that the elevated plasma levels of ir-beta EP and ir-alpha MSH post-HPD reflect the release of the intermediate lobe from tonic inhibitory dopaminergic control and (2) that, unlike the gonadotrope, hypothalamic releasing factors are not required for the maintenance of the corticotrope, or for baseline secretion of ACTH from these cells. PMID- 3029617 TI - Postpartum increases in brain oxytocin binding. AB - The binding of 3H-oxytocin in the forebrains of pregnant and postpartum rats was measured using an in vitro autoradiographic method. Binding increased selectively in the bed nucleus of the stria terminalis early in the postpartum period. This effect may be due to the physiologic changes in sex steroids late in pregnancy as the combination of ovariectomy and estrogen administration induced a similar increase in binding. PMID- 3029618 TI - Mu-receptor specificity of the opioid peptide irreversible reagent, [3H]DALECK. AB - A novel affinity reagent DALECK, i.e. D-Ala2-Leu5-enkephalin with a C-terminal chloromethyl ketone group, was previously synthesized in normal and in tritiated form and shown to react irreversibly at opioid receptors, with some evidence for selectivity for the mu subtype. DALECK tritiated in its phenolic group has been synthesized at 13-fold higher specific radioactivity than in the previous study. In the irreversible reaction of this product at pH 8.1 with rat brain membranes it was confirmed that only one polypeptide there is labelled, of apparent Mr 58,000. Competition between this reaction and ligands highly selective for the mu, delta or kappa binding sites yielded curves demonstrating the very high selectivity of the DALECK irreversible reaction for the mu site. The results provide evidence that the mu opioid receptor protein contains only one type of binding subunit, whose apparent Mr is 58,000, this size being dependent upon the conditions used in the gel electrophoresis and being higher when stringent conditions which would reduce all internal disulphide bonds are applied. PMID- 3029619 TI - Specificity of the opioid affinity reagent, DALECK, in a set of isolated tissue assay systems. AB - The C-terminal chloromethyl ketone derivative of D-Ala2-Leu5-enkephalin (DALECK) has previously been shown to act as an affinity reagent at opioid receptors. The specificity of this derivative in its reversible interaction with functional opioid receptors has been examined here in a set of four field-stimulated isolated tissue preparations; the mouse, rat and rabbit vas deferens and the guinea pig ileum. Agonist potencies relative to selective reference agonists and Schild analysis were used to elucidate the overall activity of DALECK when interacting reversibly with opiate receptors under normal physiological conditions in the isolated tissue preparations. Under these conditions the ligand shows a very strong mu-selectivity. Data obtained in the guinea pig ileum suggest that DALECK is more potent than Tyr-D-Ala-Gly-N(CH3)Phe-Gly-ol (DAGO) when acting through mu-receptors. In contrast in the mouse vas deferens DALECK is at least 70 fold less potent than the delta-ligand D-Thr2-Leu5-enkephalin-Thr (DTLET). DALECK shows little interaction with kappa-receptors. PMID- 3029620 TI - Interference of endogenous B endorphin with opiate binding in the anterior pituitary. AB - Opiates have been shown to affect prolactin secretion directly at the pituitary level. However, opiate binding sites have not been characterized on anterior pituitary tissue. In the present work we show that a soluble factor present in anterior pituitary membrane preparation inhibit opiate binding on striatal membranes. The high concentrations of B-endorphin like immunoreactivity recovered from the supernatant of pituitary membrane preparations seem to account for this effect on striatal binding and to be responsible for a partial masking of pituitary opiate receptors. We thus attempted to improve membrane fractionation procedure in order to decrease B-endorphin contamination and thus to increase binding of opiate ligands on pituitary membranes. 3-H Etorphine binds to pituitary membranes with a constant of association (kon) of 20.8 10(6) M-1 min-1 and a constant of dissociation (hoff) of 33 min-1. Saturation experiments with 3 H Etorphine and 3-H ethylketocyclazocine indicate the presence of a single population of specific binding sites (KD) value of 12.2 +/- 3.9 nM a maximal binding capacity of 55.9 +/- 10.7 fm/mg protein for 3-H Etorphine binding). These results demonstrate the presence of opiate binding sites in the anterior pituitary. However the difficulty of the membrane preparation and the low density of binding sites did not allow a complete pharmacological characterization of these sites. PMID- 3029623 TI - Immunogold demonstration of GABA in synaptic terminals of intracellularly recorded, horseradish peroxidase-filled basket cells and clutch cells in the cat's visual cortex. AB - To identify the putative transmitter of large basket and clutch cells in the cat's visual cortex, an antiserum raised against GABA coupled to bovine serum albumen by glutaraldehyde and a postembedding, electron microscopic immunogold procedure were used. Two basket and four clutch cells were revealed by intracellular injection of horseradish peroxidase. They were identified on the basis of the distribution of their processes and their synaptic connections. Large basket cells terminate mainly in layer III, while clutch cells which have a more restricted axon, terminate mainly in layer IV. Both types of neuron have a small radial projection. They establish type II synaptic contacts and about 20 30% of their synapses are made with the somata of other neurons, the rest with dendrites and dendritic spines. Altogether 112 identified, HRP-filled boutons, the dendrites of three clutch cells and myelinated axons of both basket and clutch cells were tested for the presence of GABA. They were all immunopositive. The postsynaptic neurons received synapses from numerous other GABA-positive boutons in addition to the horseradish peroxidase-filled ones. Dendritic spines that received a synapse from a GABA-positive basket or clutch cell bouton also received a type I synaptic contact from a GABA-negative bouton. A few of the postsynaptic dendrites, but none of the postsynaptic somata, were immunoreactive for GABA. The fine structural characteristics of the majority of postsynaptic targets suggested that they were pyramidal and spiny stellate cells. These results provide direct evidence for the presence of immunoreactive GABA in identified basket and clutch cells and strongly suggest that GABA is a neurotransmitter at their synapses. The laminar distribution of the synaptic terminals of basket and clutch cells demonstrates that some GABAergic neurons with similar target specificity segregate into different laminae, and that the same GABAergic cells can take part in both horizontal and radial interactions. PMID- 3029621 TI - Functional state of the pituitary-thyroid and pituitary-adrenal systems in normal pregnancy and pregnancy with late toxemia. PMID- 3029622 TI - Instantaneous inward rectification in the Mauthner cell: a postsynaptic booster for excitatory inputs. AB - Single and double electrode voltage clamp techniques have been used to analyse the leakage conductance of the goldfish Mauthner cell. The results indicate that the input conductance of this neuron is maximal at the resting potential, having an average value of 4.68 microS. Approximately 50% of this conductance is voltage dependent and could be inactivated by large hyperpolarizing command pulses (30-50 mV magnitude) of 20-35 ms duration. The magnitude and apparent time constant of the inactivation are both functions of membrane potential, such that the relaxation of the inward current increased and occurred more rapidly for greater hyperpolarizations. The presence of instantaneous inward tail currents when the imposed voltage steps were terminated and membrane potential was returned to the resting level indicated that the conductance mechanism inactivated during hyperpolarization had generated a small outward current at the resting level. Thus, we propose that a major fraction of the Mauthner cell's input conductance is a voltage-dependent K+ conductance. Hyperpolarizing inactivation is a feature of an inward K+ rectifier in other cell types, and when the recording microelectrode was located in the Mauthner cell lateral dendrite, it was possible to demonstrate characteristics consistent with rectification, namely a low conductance for outward depolarizing currents and a high conductance for inward currents. The inward rectifier of the Mauthner cell is different from that of other neurons in that it is already maximally activated at the resting potential and therefore is a major determinant of that parameter. In addition, its activation and inactivation kinetics are quite fast.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3029624 TI - In vivo presynaptic control of dopamine release in the cat caudate nucleus--I. Opposite changes in neuronal activity and release evoked from thalamic motor nuclei. AB - Halothane-anaesthetized cats implanted with three push-pull cannulae were used to estimate the effects of gamma-aminobutyric acid (GABA) application (either 10(-3) M or 10(-5) M) into the left motor nuclei of the thalamus (either ventralis medialis, or ventralis lateralis) on the firing rate of dopamine cells in the left substantia nigra (caudomedial part) and on the release of [3H]dopamine continuously synthesized from [3H]tyrosine, in the left substantia nigra (caudomedial part) and the left caudate nucleus. Preliminary experiments were performed to establish the electrophysiological characteristics of dopamine cells and non-dopamine cells in the pars compacta (mediocaudal part of substantia nigra) in groups of animals with the electrode inserted within the nigral push pull cannula or with the electrode inserted in the absence of a push-pull cannula. Dopamine and non-dopamine cells were distinguished according to several criteria (shape of the spike, duration of spike, frequency of discharge, conduction velocity estimated following antidromic activation from the caudate nucleus for dopamine cells or from the ventralis medialis for non-dopamine cells). Data obtained from recordings made within the push-pull cannula were identical to those obtained in the absence of the cannula. In addition both the intravenous injection of amphetamine or its local application (10(-6) M) in the substantia nigra inhibited the firing rate of dopamine cells. When GABA was applied at 10(-3) M for 30 min into the ventralis medialis-ventralis lateralis the multi-unit activity of thalamic cells recorded within the push-pull cannula was inhibited. Single unit activity of dopamine cells was also inhibited and [3H]dopamine release was reduced in the caudate nucleus and increased in the substantia nigra. These results suggest that under these conditions, dopamine release from nerve terminals depended upon nerve activity and that dopamine released from dendrites inhibited the activity of dopamine cells. When GABA was applied at 10(-5) M for 30 min into the ventralis medialis-ventralis lateralis, multi-unit activity of thalamic cells was increased, single-unit activity of dopamine cells was inhibited and [3H]dopamine release was enhanced in the ipsilateral caudate nucleus and not affected in the left substantia nigra, demonstrating that in this situation the release of dopamine from nerve terminals was not dependent on the firing rate of dopamine cells. In addition, these results indicated that the activity of dopamine cells was not always dependent on the dendritic release of dopamine.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3029625 TI - Synaptic connections, axonal and dendritic patterns of neurons immunoreactive for cholecystokinin in the visual cortex of the cat. AB - A subpopulation of gamma-aminobutyric acid (GABA) containing neurons was reported to contain cholecystokinin-immunoreactive material in the visual cortex of cat [Somogyi et al., J. Neurosci. (1984) 4, 2590-2603]. In the present study pre embedding immunocytochemistry was used to identify which of the several types of presumed GABAergic nonpyramidal cells in areas 17 and 18 contain cholecystokinin immunoreactivity. Most of the cholecystokinin-immunoreactive somata were found in layers II-III, they were less frequent in layers I and VI, and relatively rare in layers IV and V. The distribution and density of the axon terminals resembled that of the cell bodies. Two well defined types of cholecystokinin-immunoreactive neuron were distinguished: (1) double bouquet cells in layers II-III with vertically projecting axons, and (2) small basket cells with local axons either restricted to layers II-III, or descending to layer V. Additional cholecystokinin positive cells showed features of bitufted or multipolar neurons in layers II-VI and horizontal cells in layer I, but these cells could be defined less well due to partial staining. Cholecystokinin-immunoreactive dendrites were found to run horizontally in layer I for several hundred micrometers. Some of the cholecystokinin-immunoreactive cells in layer VI had very long dendrites ascending radially up to layer III, as did their axons. A few cholecystokinin immunoreactive cells appeared to have two axons and this was confirmed by electron microscopy. All cholecystokinin-immunoreactive neurons and terminals were separated from the basal lamina of blood vessels by glial endfeet. Random samples of boutons from each layer as well as identified terminals traced to their origin from local neurons were examined in the electron microscope. All of the boutons established symmetrical (type II) synaptic contacts with dendritic shafts, spines or somata. Quantitative electron microscopy of the postsynaptic targets of double bouquet cells and small basket cells demonstrated clear differences between these two types of neuron; basket cells having a higher proportion of their terminals in synaptic contact with somata. The findings that several distinct types of cortical neurons, as defined by their synaptic connections, contain cholecystokinin-immunoreactive material and that identified axons of all examined neurons form type II synaptic contacts suggests that the majority, if not all cholecystokinin-positive boutons forming type II contacts originate from local cortical cells. The distribution of targets postsynaptic to cholecystokinin-positive neurons is compared to those of cells labelled by other methods. PMID- 3029626 TI - N-methylaspartate receptors mediate epileptiform activity evoked in some, but not all, conditions in rat neocortical slices. AB - Using intracellular recordings from pyramidal neurons in isolated slices of rat cerebral cortex epileptiform discharges evoked (1) in the presence of gamma aminobutyric acid antagonists, and (2) in the absence of Mg2+ were compared. Depolarization shift responses recorded in the presence of bath applied picrotoxin, or electrophoretically applied picrotoxin or bicuculline, were similar in many respects to depolarization shifts reported previously, except that they could be evoked by stimuli subthreshold for evoking discernible postsynaptic potentials in these experiments. Large depolarizations evoked by repetitive activation of an N-methylaspartate receptor mediated synapse in the absence of Mg2+, displayed several properties similar to those of depolarization shifts evoked in the presence of gamma-aminobutyric acid antagonists, i.e. similar shape, latency, inability to follow high repetition rates and a similar voltage relation, suggesting activation of the same cellular mechanism. "Slow spikes" evoked as part of the response to electrophoretically applied N methylaspartate were augmented, i.e. they were replaced by larger, longer, more complex events, when gamma-aminobutyric acid antagonists were applied. The potentiated response, evoked in the absence of Mg2+, was dependent on the activation of an N-methylaspartate receptor mediated synapse and was blocked by N methylaspartate antagonists. In contrast, depolarization shifts could be evoked in the presence of large doses of N-methylaspartate antagonists, when gamma aminobutyric acid antagonists were applied. Spontaneous depolarizations similar to depolarization shifts were recorded when cells were exposed to low, tonic, electrophoretic applications of excitatory amino acids under control conditions. In addition, some potentiation of the N-methylaspartate receptor mediated excitatory postsynaptic potential was achieved in the presence of Mg2+ when cells were depolarized by 10-20 mV. Depolarization shifts evoked when bicuculline was applied electrophoretically to different parts of the dendritic field, some hundreds of microns from the soma, differed in shape, latency and time course and the depolarization shift evoked when bicuculline was applied at one site summed with the depolarization shift evoked when it was applied elsewhere. We conclude that different inputs are required to activate the responses evoked in the presence of gamma-aminobutyric acid antagonists and in the absence of Mg2+. The possibility that both involve activation of dendritic Ca2+ currents and that the magnitude of the response depends on the proportion of the dendritic field activated, is discussed. PMID- 3029627 TI - Synaptic connections of axo-axonic (chandelier) cells in human epileptic temporal cortex. AB - The human temporal cortex contains a type of interneuron, identified by Golgi impregnation which, like the axo-axonic or chandelier cells found in animals, establishes Gray's type II synaptic contacts exclusively with the axon initial segments of pyramidal cells. Each terminal segment is composed of 3-12 boutons to form a "chandelier"-like appearance. For the two human axo-axonic cells analysed in this study we could identify 269 and 86 bouton rows respectively, which represents an equivalent number of postsynaptic pyramidal cells. A terminal bouton row from one of these Golgi-impregnated cells was shown to be in synaptic contact with the axon initial segment of a Golgi-impregnated pyramidal cell. The very specific nature of the target of axo-axonic cells, together with their highly divergent axonal arborization, means that they are ideally placed to control the output of a large population of pyramidal cells. Since previous studies in animals have shown the GABAergic nature of axo-axonic cells it is possible that human axo-axonic cells could be involved in the generation of epileptic activity or in the control of its propagation. PMID- 3029629 TI - Case for diagnosis. Avian pox. PMID- 3029628 TI - Cerebrospinal fluid concentrations of corticotropin-releasing hormone (CRH) and corticotropin (ACTH) are reduced in patients with Alzheimer's disease. AB - We examined corticotropin-releasing hormone-like immunoreactivity (CRH-LI) and corticotropin (ACTH) levels in the CSF of 33 patients with presumptive Alzheimer's disease (AD) and 13 healthy, age-matched controls. The mean CRH-LI and ACTH levels of the AD patients were significantly less than controls. Despite these reductions, none of the patients had evidence of pituitary-adrenal dysfunction. A disorder of extrahypothalamic CRH may be involved in the pathophysiology of AD. PMID- 3029630 TI - [Clinico-biological analysis of 192 cases of breast cancer]. PMID- 3029631 TI - [Gastroduodenal and ileal adenomas in familial diffuse polyposis. Therapeutic prospectives]. PMID- 3029632 TI - [Clinico-radiologic aspects of calcium pyrophosphate dihydrate deposition disease]. AB - Calcium pyrophosphate dihydrate crystal deposition disease is a clinical condition characterised by Gout-like synovitis (pseudogout), calcification on and around the joints and an arthropathy that is radiologically similar to osteoarthritis (chronic pyrophosphate arthropathy). Though all these radiological clinical aspects may coexist in the same patient this is often not the case. An examination of the X-ray data on the 68 cases studied which were diagnosed on the basis of the criteria proposed by McCarty, shows that the disease is relatively common especially in the over-fifties. When chronic pyrophosphate arthropathy is the only clinical manifestation of the disease differential diagnosis from the osteoarthrosis so common in the elderly is difficult and depends on the greater severity and progression of the joint damage that may often affect joints not subjected to weight such as the shoulder, unlike what happens in osteoarthritis. PMID- 3029633 TI - Tensor tympani reflex pathways studied with retrograde horseradish peroxidase and transneuronal viral tracing techniques. AB - The neural pathway involved in activation of the tensor tympani (TT) muscle was studied in the rat using retrograde HRP and transneuronal viral tracing techniques. The pool of TT motoneurons labeled with HRP was located ipsilaterally under the anterior third of the trigeminal motor nucleus and extended rostrally towards the lateral lemniscus. The origin of the inputs to these motoneurons was then determined using transneuronal viral transport: presumably transneuronally infected neurons appeared bilaterally in the vicinity of the superior olivary complex, mainly in between the two nuclei of the trapezoid body. The present data are consistent with previous conclusions based on lesion experiments that the TT reflex loop is made up of a chain of 4 neurons. PMID- 3029634 TI - Schwann cell galactocerebroside of unmyelinated fibers is inducible by derivatives of adenosine 3',5'-monophosphate. AB - By using indirect immunofluorescence, galactocerebroside (galC) was detected on the surface of Schwann cells cultured from unmyelinated fibers of 10- to 12-day old rat cervical sympathetic nerve trunk. By day 4 in vitro, galC-positive cells disappeared from the culture. When the 4-day cultures were treated with 1 mM 8 bromo cyclic AMP or dibutyryl cyclic AMP, galC reappeared in 72 h. The proportion of Schwann cells expressing galC was dependent on the concentration of cyclic AMP derivatives used. PMID- 3029635 TI - Astrocyte glutamate receptor activation promotes inositol phospholipid turnover and calcium flux. AB - Astrocyte-enriched cultures prepared from the neonatal rat cortex were prelabelled with either [3H]myoinositol or 45Ca2+ and then exposed to various excitatory amino acids. This resulted in an increase in both the breakdown of membrane inositol phospholipids and Ca2+ flux with the following rank order of efficacy: quisqualate greater than or equal to glutamate (Glu) greater than kainate much greater than N-methyl-D-aspartate. Experiments performed with the Ca2+ ionophore A23187 and in the absence of medium Ca2+ suggested that Glu-evoked 45Ca2+ efflux was primarily the result of an increased influx of extracellular Ca2+. However, Glu-stimulated inositol lipid metabolism was found to be only partially dependent on extracellular Ca2+. The quisqualate-preferring receptor antagonist gamma-glutamylaminomethylsulphonic acid was found to be effective in reversing both Glu-evoked inositol lipid breakdown and Ca2+ flux. The results presented are suggestive of some form of interaction between Glu receptors coupled to inositol lipid turnover and Ca2+ channel opening in astrocytes. PMID- 3029636 TI - Muscle relaxant and anticonvulsant activity of 3-((+/-)-2-carboxypiperazin-4-yl) propyl-1-phosphonic acid, a novel N-methyl-D-aspartate antagonist, in rodents. AB - A novel 4-substituted derivative of piperazine-2-carboxylic acid, 3-((+/-)-2 carboxypiperazin-4-yl)-propyl-1-phosphonic acid (CPP), with potent N-methyl-D aspartate (NMDA) antagonist activity was evaluated as a muscle relaxant in genetically spastic rats. CPP, 0.02-0.1 mmol/kg, given intraperitoneally reduced the tonic activity in the electromyogram recorded from the gastrocnemius muscle of genetically spastic rats in a dose- and time-dependent manner. Muscle relaxation was also seen following intrathecal application of CPP, 0.0002-0.002 mumol, in genetically spastic rats. CPP, 0.1 mmol/kg, while not affecting Hoffman (H-) reflexes, depressed flexor reflexes in anesthetized rats following intravenous administration. In mice, CPP, 0.001 mumol, given intracerebroventricularly preferentially antagonized myoclonic seizures induced by NMDA and quinolinate, and had no effect on convulsions elicited by kainate, quisqualate and L-glutamate. These observations identify CPP as the most potent preferential NMDA antagonist so far tested with muscle relaxant and anticonvulsant activity resembling the profile of action of 2-amino-7 phosphonoheptanoate. PMID- 3029637 TI - Internal presynaptic tetraethylammonium (TEA+) blocks cholinergic transmission at a synapse between identified neurones. AB - Intracellular microelectrodes were used to study a cholinergic synapse between two identified neurones: the lateral filiform hair sensory neurone (LFHSN) and giant interneurone 3 (GI 3) in the terminal ganglion of the first-instar cockroach Periplaneta americana. The presynaptic neurone (LFHSN) was impaled in a region of the axon which forms large numbers of output synapses onto GI 3. Intracellular injection of tetraethylammonium (TEA+) into LFHSN blocked LFHSN-GI 3 synaptic transmission. Injection of TEA+ and either acetylcholine (ACh) or choline into the axon preserved synaptic transmission. TEA+ may compete with choline at an intracellular site involved in the maintenance of releaseable ACh. PMID- 3029639 TI - Lead neuropathy and electrophysiological studies in low level lead exposure: a critical review. AB - The literature on nerve conduction velocity (NCV) studies and electromyography in lead exposed populations was reviewed. All studies revealed some degree of defect in their experimental design. Disregarding these limitations, a consensus of the studies indicates that a mild slowing (7%) of motor and sensory NCV may be present in the median and posterior tibial nerves, but that ulnar and peroneal NCVs are not slowed in subjects with blood lead levels of less than 60 micrograms/dl. No clinical correlations to this mild slowing were reported in studies where concurrent motor and sensory examinations were performed. A majority of the studies did not find a correlation between blood lead level and NCV at blood lead levels below 70 ug/dl or length of exposure and NCV. Most studies of electromyographic activity in lead exposed population found abnormalities only in patients with blood lead levels above 60-70 ug/dl or in patients with lead colic or other systemic symptoms of lead intoxication. This review does not find evidence that the slowing of NCV in low level lead exposure is clinically significant or evidence that would suggest that it is a subclinical manifestation of clinical lead neuropathy in man. PMID- 3029638 TI - [3H]TCP binding sites in Alzheimer's disease. AB - Quantitative autoradiography was used to determine the density and distribution of [3H]1-[1-(2-thienyl)-cyclohexyl]piperidine ([3H]TCP) binding sites in human hippocampal tissue sections from control and Alzheimer's disease patients. Some Alzheimer's cases showed no changes in binding site density while other cases showed substantial declines in the CA1 region. [3H]TCP binding in the CA1 region from Alzheimer's patients was reduced an average of 40% while the other hippocampal regions were unaffected. It is proposed that the loss of [3H]TCP sites in the hippocampal CA1 region of certain Alzheimer's cases is associated with the greater cell loss observed in cases of severe Alzheimer's disease. PMID- 3029640 TI - A liver cell receptor for uptake of free fatty acids. PMID- 3029641 TI - Metastatic gestational trophoblastic disease: experience at the New England Trophoblastic Disease Center, 1965 to 1985. AB - This report reviews the results of therapy in 93 patients with metastatic gestational trophoblastic tumor treated from 1965-1985. Complete remission was achieved in all 42 patients with low-risk metastatic disease and in 34 of 51 patients (67%) with high-risk metastatic disease. Single-agent chemotherapy induced complete remission in 38 of 42 patients (91%) with low-risk metastatic disease. Survival of high-risk patients has improved markedly over the past two decades; complete remission was attained in 13 of 24 high-risk patients (54%) from 1965-1975, and in 21 of 27 (78%) from 1976-1985. Survival correlated with the number of high-risk factors, the prognostic score, and the type of treatment. From 1965-1975, 54% (13 of 24) of high-risk patients were treated with single agent chemotherapy alone, while in the last decade only 7% (two of 27) were so treated. Twenty-one patients with traditional high-risk factors had a prognostic score of 7 or less, and all achieved remission, with 67% (14 of 21) treated with primary single-agent chemotherapy. The prognostic scoring system was more effective than traditional high-risk criteria at predicting which patients require intensive combination chemotherapy to attain remission. PMID- 3029642 TI - Human papillomaviruses in women with a history of abnormal Papanicolaou smears and in their male partners. AB - Human papillomavirus infection of the genital tract was identified by the filter in situ hybridization test. Exfoliated cervical cells were tested separately for the prevalence of human papillomavirus 6/11 and 16/18. Human papillomavirus deoxyribonucleic acid (DNA) was identified in 70 and 92% of specimens of U.S. and West German women, respectively, who showed concurrent cytologic and colposcopic abnormalities, and in 50 and 54% of women, respectively, who showed neither cytologic nor colposcopic abnormalities at the time of examination. In the cytologic categories of condyloma, mild to moderate dysplasia (cervical intraepithelial neoplasia I/II), and severe dysplasia-carcinoma in situ (cervical intraepithelial neoplasia III), the overall DNA detection rate of human papillomavirus 6/11 and 16/18 varied between 75 and 83%; but human papillomavirus 16/18 was recovered relatively more frequently from the more severe lesions. Forty-eight West German women were monitored cytologically over a period of three to 24 months; progression to carcinoma in situ (cervical intraepithelial neoplasia III) was correlated with initial isolation of human papillomavirus 16/18. The vagina and vestibule were found to be frequent sites of human papillomavirus infection with the same virus type as in the cervix. In an investigation of male partners of 40 human papillomavirus-positive women, human papillomavirus was identified in exfoliated cells from 26; in 19 instances, the males harbored the same human papillomavirus types as their female partners. PMID- 3029643 TI - Inter- and intra-pathologist variability in the diagnosis of gestational trophoblastic neoplasia. AB - All 190 cases of gestational trophoblastic neoplasia diagnosed in the Baltimore metropolitan area from 1975-1982 were identified. Histologic slides were requested and reviewed independently by two pathologists who agreed upon uniform criteria for the diagnosis of hydatidiform (complete) mole, invasive mole, and choriocarcinoma. A representative sample of the slides was selected and resubmitted to one of the study pathologists for a second review. The inter- and intra-pathologist variability in the diagnosis of gestational trophoblastic neoplasia was calculated using the kappa statistic (K). Our findings indicated that the variability in the diagnosis of gestational trophoblastic neoplasia was low whereas that for the related tumor of incomplete mole was high. PMID- 3029644 TI - [Effect of therapy with cytostatic drugs on the hemostasis system in patients with small cell and non-small cell bronchial cancers, malignant lymphomas and plasmacytomas]. AB - Hemostatic analyses were carried out on 43 patients with small and non-small cell lung cancer, malignant lymphomas and plasmacytomas prior to, 4 h and 24 h after application of chemotherapy. The fibrinopeptide A (FPA) level and the FPA in vitro generation rate (delta FPA) increased significantly only in the small cell lung cancer and malignant lymphomas after 4 h. This supports the thesis of the clotting activation after cytostatic-induced exposition of the tumour cell thromboplastins. An increase in the FPA was observed in those tumor patients, where a high therapy-induced cell destruction was expected. An increase in the activity of factor VIII, as was seen in acute phase reactions, also led to hypercoagulability. Fibrinogen and plasminogen decreased significantly after chemotherapy. Evidence of relevant contact activation was not found in the missing deviation of the C1 inhibitor. The increase in protein C may possibly be attributed to the high-dose corticoid therapy. PMID- 3029645 TI - [Clones of the lymphoblastoid line RPMI-6410t requiring an exogenous growth factor]. AB - It was earlier established that the RPMI-6410t cells, obtained from a patient with acute myeloblastemia, synthesized a growth factor which maintains their proliferation and had a specific receptor for this factor on their surface. The use of a medium conditioned by the 6410t cells made it possible to define conditions in which practically 100% efficiency cloning of these cells is attained by the method of limiting dilutions. In the present work, this method of cloning was applied to obtain from the 6410t strain clones which are characterized by a requirement for an exogenous growth factor. These clones, like the 6410t cells, have on their surface specific receptors but, unlike the parental cells, do not synthesize the growth factor and do not form colonies in soft agar, i. e. lose one of the features of malignancy. These facts agree with the published data according to which the proliferation of normal cells is regulated by exogenous growth factors and confirm a suggestion put forward in our previous work that the endocrine regulation of cell growth is one of the mechanisms of malignancy. PMID- 3029646 TI - Klippel-Trenaunay-Weber syndrome. AB - Klippel-Trenaunay-Weber syndrome exhibits vascular anomalies including hemangiomas and varicose veins that commonly appear in the facial area. Characteristic findings involving the oral cavity include an enlarged maxilla, displacement of teeth, and malocclusions. Two cases are presented, with generalized and oral findings ranging from a mild to a very severe form of the syndrome. PMID- 3029648 TI - [Biochemical evaluation of the intensity of post-traumatic inflammation after injuries of the joints of the lower extremities]. PMID- 3029647 TI - Adenosquamous carcinoma of the floor of the mouth and lower alveolus: a radiation induced lesion? AB - A case of adenosquamous carcinoma of the floor of the mouth and alveolus that occurred following radiation therapy is described. The possible role of radiation in the etiology of this lesion is discussed, and the complex histopathologic features of this neoplasm are emphasized. PMID- 3029649 TI - [Detection of mixed rotavirus infections by the viral RNA test]. PMID- 3029651 TI - Reduction of niridazole by metronidazole resistant and susceptible strains of Trichomonas vaginalis. AB - The inhibitory effect of niridazole on hydrogen production by metronidazole resistant (CDC-85) and susceptible (C1-NIH) Trichomonas vaginalis strains was investigated. The results show that niridazole is more effective than metronidazole in inhibiting hydrogen production by the resistant isolate. In CDC 85 aerobic inhibition requires a 4-fold increase in metronidazole concentration compared with that required anaerobically, but the corresponding factor for niridazole is only 1.5-fold. Reduction of the drug by a hydrogenosome-enriched preparation gave rise to a multiline electron spin resonance detectable signal, which is due to a nitrogen-centred radical. PMID- 3029650 TI - Biochemical and ultrastructural investigation of the effect of Stelazine (trifluoperazine) on Hymenolepis diminuta (Cestoda). AB - The effects of the phenothiazine, Stelazine, on Hymenolepis diminuta were investigated. The cestode was incubated for 10 min at 37 degrees C with 1 mM trifluoperazine, in the presence and absence of Ca2+. Assay of brush border enzymes showed that drug treatment lowered the activities of alkaline phosphatase, Ca2+-ATP'ase, 5'-nucleotidase and type 1 phosphodiesterase. This occurred in parallel with a significant reduction in tegumental protein. Under these conditions gross changes in ultrastructural appearance and cellular organization were observed. There was a lack of ordered microtriches and the distal cytoplasm was absent. Glycogen granules were scattered throughout the cytoplasm within the subtegumental layer. The connective tissue also appeared to be in some disarray. The effects of Stelazine appeared to be dependent on time and were significantly increased when Ca2+ was included in the incubation medium. Incubation with the less hydrophobic phenothiazine trifluoperazine sulphoxide had minimal effect on the integrity of the cestode. The results reported here support the premise that certain phenothiazines may be considered as potential cestocidal agents. PMID- 3029652 TI - [The Prophy-jet in dental practice]. PMID- 3029653 TI - Comparison of peroxidase-antiperoxidase and avidin-biotin complex methods for the detection of papillomavirus in histological sections of the cervix uteri. AB - A study was undertaken to determine the relative sensitivities of the peroxidase antiperoxidase (PAP) and avidin-biotin complex (ABC) methods for the detection of human papillomavirus (HPV) antigens in acetic acid-ethanol fixed paraffin embedded cervical tissue. Tissue sections prepared from 14 women suspected to have HPV infections with either atypia or dysplasia were stained immunohistochemically using an antiserum against genus-specific (common) antigen of bovine papillomavirus. Detection of HPV antigen was approximately twice as frequent by the ABC method as by the PAP method. Of the 14 cases studied, 43% were found to be HPV positive by the PAP method whereas 79% were HPV positive by the ABC method. In addition, the number of cells found to be HPV positive by the ABC method was approximately double the number by the PAP method. PMID- 3029654 TI - [Cytodiagnosis of early stomach cancer]. PMID- 3029655 TI - [Small-cell carcinoma of the endometrium with glandular and squamous differentiation]. PMID- 3029656 TI - Primary carcinoma of the trachea: combined small cell, squamous cell and giant cell carcinoma. Report of a case and a review of the literature. PMID- 3029657 TI - Beta-adrenergic receptors and cyclic adenosine monophosphate generation in human fetal lung. AB - We designed experiments to determine whether beta-adrenergic receptors are present and functional in human fetal lung during the 2nd trimester of gestation. To determine the presence of beta receptors, characterize their binding sites, and assess changes in receptor with gestational age, we performed radioligand binding assays with the specific, high-affinity beta antagonist, 125I iodocyanopindolol, in membrane particulates from the lungs of 2nd trimester abortuses (15-23 wk). Binding of 125I-iodocyanopindolol was saturable and of high affinity (dissociation constant = 40 pM). Binding was stereoselective as determined by competition studies with (-) and (+) stereoisomers of propranolol. Agonist affinities (isoproterenol greater than epinephrine much greater than norepinephrine) were consistent with a predominance of beta-2 receptors; this predominance was confirmed by competition studies with the specific beta-2 receptor antagonist ICI 118-551 (75% beta-2, 25% beta-1). The concentration of beta-adrenergic receptors increased with gestational age. To assess the functional coupling of the beta receptors, we tested the ability of receptor occupancy to activate adenylate cyclase. For this assay, we incubated minced human fetal lung with beta agonists and determined the amount of cAMP generated. beta Agonists stimulated cAMP generation more than 2-fold. We conclude that beta adrenergic receptors are present and functional in human fetal lung as early as the 2nd trimester. PMID- 3029658 TI - Increased activity of the respiratory burst in cord blood neutrophils: kinetics of the NADPH oxidase enzyme system in subcellular fractions. AB - Previous studies with neutrophils from newborn infants compared to neutrophils from healthy adults have documented increased respiratory burst activity including enhanced superoxide anion (O2-) production, nitroblue tetrazolium dye reduction, and hexose monophosphate shunt activity. To investigate the biochemical basis for these observations, we examined oxidative metabolism in membrane-rich fractions of neutrophils. Neutrophils from cord blood of vaginally delivered term infants or healthy adults were disrupted by nitrogen cavitation and subcellular fractions collected on discontinuous sucrose density gradients. Subcellular fractions of newborn neutrophils separated in a fashion identical with samples from healthy adults. Activity of alkaline phosphatase, a plasma membrane marker, was increased 4- to 5-fold in disrupted cells free from nuclei (postnuclear supernatant) as well as plasma membrane fractions from newborn samples compared to those from healthy adults. Content of lactoferrin, a specific granule marker, was decreased in postnuclear supernatants but equivalent in specific granule fractions of newborn cells compared to those from adults. No differences were noted in myeloperoxidase content of postnuclear supernatants or any other subcellular fraction. Plasma membrane fractions from phorbol myristate acetate-stimulated cord blood neutrophils made significantly more O2- than samples from adults (newborn 32.9 +/- 8.1 nmol O2-/min/mg protein mean +/- SEM, n = 3 versus adult 10.8 +/- 4.2, n = 3; p less than 0.05). Plasma membrane-rich fractions were also collected by the technique of differential centrifugation and kinetic parameters of the NADPH-dependent oxidase enzyme(s) were measured for vaginally delivered newborn and adult samples.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3029659 TI - Oral therapy with acyclovir in infants and children. PMID- 3029660 TI - Foodborne Snow Mountain agent gastroenteritis in a school cafeteria. AB - In 1984, an outbreak of gastroenteritis occurred at a school with 1,860 students in Brooklyn, NY. In a single-stage cluster sample of 375 students, 129 (34%) had illnesses that met our case definition of vomiting or diarrhea. The mean incubation period was 26 hours, and the mean illness duration was 24 hours. All case students had eaten in the cafeteria on at least one day between Nov 13 and 16, compared with 174/214 (81%) noncase students (P = 10(-8), Fisher exact test). Foods implicated were french fries (relative risk 1.7, 95% confidence limits 1.4, 2.0) and hamburgers (relative risk 1.6, 95%, confidence limits 1.2, 2.1). Two cafeteria employees had served those foods while affected by diarrhea. By a recently developed blocking enzyme-linked immunosorbent assay, six of 11 (55%) case students showed fourfold antibody increases between acute- and convalescent phase serum samples for Snow Mountain agent, a Norwalk-like virus, compared with one of ten (10%) noncase students (P = .04, Fisher exact test). We strongly suspect, but cannot document conclusively, that the Snow Mountain agent was spread to students on a vector of hot foods contaminated by ill food handlers. Implicated foods conferred low relative risks and could only have accounted for 74% of cases of illness. The strong association between cafeteria exposure and illness, therefore, suggests that additional modes of spread occurred. PMID- 3029661 TI - Health supervision visits: should the immunization schedule drive the system? PMID- 3029662 TI - Electron microscopy for diagnosis of tumors in children. PMID- 3029663 TI - The contribution of tissue culture to the study of solid tumors of childhood. PMID- 3029664 TI - [Clinical research on the demonstration of the portal venous system and its collaterals using dynamic images of liver scintigraphy]. PMID- 3029665 TI - [Gallbladder abnormalities following transcatheter arterial embolization of hepatic malignancies]. PMID- 3029666 TI - Reciprocal recombination products of VK-JK joining reactions in human lymphoid cell lines. AB - The recombination process that joins a VK to a JK segment of an immunoglobulin gene generates a second, reciprocal recombination product called f fragment. In this second product the regions flanking the VK and JK segments in the germline are joined in a head to head fashion. We now analysed f fragments in the human lymphoid cell lines Daudi, JI and IARC/BL41. All three f fragments contain JK1 flanks; the VK derived moiety of f Daudi and f41 could be traced back to known germline VK genes. There is a precise head to head joining of the heptanucleotide signal sequences in f Daudi and fJI while in f41 six nucleotides are present between the signal sequences. In contrast to the VK-JK recombination products, the f fragments were found to lack somatic mutations. The structures of the f fragments are discussed in the context of the VK-JK rearrangement mechanism. PMID- 3029667 TI - The control of lambda DNA terminase synthesis. AB - Nu1 and A, the genes coding for bacteriophage lambda DNA terminase, rank among the most poorly translated genes expressed in E. coli. To understand the reason for this low level of translation the genes were cloned into plasmids and their expression measured. In addition, the wild type DNA sequences immediately preceding the genes were reduced and modified. It was found that the elements that control translation are contained in the 100 base pairs upstream from the initiation codon. Interchanging these upstream sequences with those of an efficiently translated gene dramatically increased the translation of terminase subunits. It seems unlikely that the rare codons present in the genes, and any feature of their mRNA secondary structure play a role in the control of their translation. The elimination of cos from plasmids containing Nu1 and A also resulted in an increase in terminase production. This result suggests a role for cos in the control of late gene expression. The terminase subunit overproducer strains are potentially very useful for the design of improved DNA packaging and cosmid mapping techniques. PMID- 3029668 TI - Tsetse fly rDNA: an analysis of structure and sequence. AB - A genomic library of Glossina morsitans morsitans (tsetse fly) has been constructed in the phage vector EMBL 4 and a complete rDNA unit isolated by using a D. melanogaster rDNA clone as a probe. The overall organisation is typical of higher eukaryotes, including an intergenic spacer consisting of a subrepeating structure. Atypically, however, the 45S precursor RNA promoter was shown to lie within the last subrepeat by S1 mapping; i.e. the last subrepeat extends 90 bp into the ETS. The sequence of the spacer subrepeats, the ETS and the first 151 nucleotides of the 18S gene was determined. Comparisons with the corresponding regions of other higher eukaryotes, including insects shows that the ETS has completely diverged, raising questions concerning their functional significance and evolutionary retention; depending on the method of alignment, only two short regions of reasonable homology are shared with Drosophila species: a stretch of nucleotides around the transcription initiation site, and AACATA at the NTS-18S gene junction; and the functionally important G at -16, conserved in all other examined species, is displaced no matter what method of alignment is used. These and other features reflect continual processes of change in the rDNA family to which the several functions of the repeating unit need to adjust. PMID- 3029669 TI - The pro alpha 2(V) collagen gene is evolutionarily related to the major fibrillar forming collagens. AB - A number of overlapping cDNA clones, covering 5.2 kb of sequences which code for the human pro alpha 2(V) collagen chain, have been isolated. Analysis of the structural data have indicated a close evolutionary kinship between the pro alpha 2(V) chain and the major fibrillar collagen types. Isolation and analysis of an 8 kb genomic fragment has further supported this notion by revealing a homologous arrangement of nine triple-helical domain exons. These studies have therefore provided conclusive evidence which categorizes the Type V collagen as a member of the Group 1 molecules, or fibrillar-forming collagens. PMID- 3029670 TI - Alternative 5' exons either provide or deny an initiator methionine codon to the same alpha-tubulin coding region. AB - The primary structures of two overlapping novel alpha-tubulin cDNA clones isolated from a Macaca fascicularis testis cDNA library and the corresponding human gene are presented. Although the general structure of the human gene conforms to that of previously described mammalian alpha-tubulin genes, there is a surprising difference: the ATG initiator codon is conspicuously absent. The macaque testis cDNA similarly lacks the initiator methionine, but otherwise encodes a variant alpha-tubulin isotype precisely conserved in the human gene. RNA blot analysis in the macaque, using a 3' untranslated region probe, revealed the existence of two additional related transcripts expressed in every tissue examined except the adult testis. Sequence comparisons indicate that the 2.0 kb testis transcript and one of the additional transcripts result from differential transcription of the same gene. The two transcripts differ only at the 5' end as a result of the recruitment of different 5' exons. Curiously, the 5' exon utilized outside the testis encodes an initiator methionine in the expected location. PMID- 3029671 TI - Synthesis of decadeoxyribonucleotides containing N6-methyladenine, N4 methylcytosine, and 5-methylcytosine: recognition and cleavage by restriction endonucleases (nucleosides and nucleotides part 74). AB - The naturally-occurring modified bases, N6-methyladenine, N4-methylcytosine, and 5-methylcytosine were chemically introduced in place of the adenine or cytosine in the decadeoxyribonucleotides containing recognition sequences of Bgl II, Sau 3AI, Mbo I and Mfl I. The modified oligomers bind to the enzymes but the rates of cleavage by the enzymes are variable. PMID- 3029672 TI - The sequence of the Saccharomyces cerevisiae gene PHO2 codes for a regulatory protein with unusual aminoacid composition. AB - A new centromere vector for the construction of a Saccharomyces cerevisiae gene library, allowing direct selection for DNA insert, will be described. From that library the gene for the regulatory protein PHO2 involved in PHO5 induction has been cloned by complementation of a pho2 mutation. The complementing activity was shown to be located on a 3.6 kb HindIII fragment. This fragment was used to evict the genomic copy and with appropriate genetic crosses we proved, that the cloned gene is PHO2. The DNA sequence of PHO2 was determined. Analysis of the sequence data uncovered striking homology regions with PHO4, another protein necessary for the induction of PHO5. The relevance of the observed homology will be discussed. PMID- 3029673 TI - Temperature and salt dependence of the gel migration anomaly of curved DNA fragments. AB - A series of oligonucleotides of different sequences have been cloned to study DNA curvature. Several DNA fragments containing these oligonucleotides in various numbers of repeats were analyzed in 10% polyacrylamide gels. A strong gel migration anomaly was found for dA4 sequences; a comparably very small but clearly detectable anomaly was observed for dA3 (both in a repeat length of 10 base-pairs). The temperature and salt (NaCl, MgCl2) dependence of the gel migration anomaly of these DNA fragments was measured. While a similar behaviour of all sequences is observed for the addition of NaCl, the temperature and MgCl2 dependence of the anomaly varies with the oligonucleotide sequence. These data are interpreted in terms of local DNA structure changes induced by changes in the temperature and the MgCl2 concentration which affect the planarity of the curved DNA fragments. PMID- 3029675 TI - Nucleotide sequence and expression of the gene encoding the EcoRII modification enzyme. AB - The gene coding for the EcoRII modification enzyme has been cloned and the nucleotide sequence of 1933 base pairs containing the gene has been determined. The gene codes for a protein of 477 amino acids. Two transcriptional start sites have been mapped by S1 mapping. One deletion that removes 34 N-terminal amino acids was found to have partial enzyme activity. Comparison of the EcoRII methylase sequence with other cytosine methylases revealed several domains of partial homology among all cytosine methylases. Cloning the gene in multicopy pUC vectors increased the expression by 6-18 fold. A 40 fold overproduction of the EcoRII methylase was obtained by cloning the gene in the expression vector carrying the lambda PL promoter. PMID- 3029677 TI - Determination of size and orientation of DNA fragments cloned in phage M13 by S1 nuclease mapping. PMID- 3029676 TI - Structural analysis of the mouse chromosomal gene encoding interleukin 4 which expresses B cell, T cell and mast cell stimulating activities. AB - Based on homology with the mouse interleukin 4 (IL-4) cDNA that expresses B cell, T cell, and mast cell stimulating activities (Lee, F. et al., (1986) Proc. Natl. Acad. Sci. USA 83, 2061), we have isolated from a Balb/c mouse liver DNA library the mouse chromosomal gene and analyzed its overall structure. The gene occurs as a single copy in the haploid genome and contains four exons and three introns. The exon sequences almost completely match the cloned cDNA sequence. Interestingly, there is a fairly high degree of homology between mouse IL-4 and mouse IL-2 genes extending more than 200 bp upstream of a "TATA" like sequence located 20 bp upstream of the transcription initiation site. These sequences may play an important role in the regulated expression of this gene in concanavalin A or antigen-stimulated T lymphocytes. The supernatant of COS7 cells transfected with plasmid DNA containing the entire gene exhibited both T cell growth factor and mast cell growth factor activities. PMID- 3029674 TI - Organization and partial sequence of a DNA region of the Rhizobium leguminosarum symbiotic plasmid pRL6JI containing the genes fixABC, nifA, nifB and a novel open reading frame. AB - By hybridization and heteroduplex studies the fixABC and nifA genes of the Rhizobium leguminosarum symbiotic plasmid pRL6JI have been identified. DNA sequencing of the region containing nifA showed an open reading frame of 1557 bp encoding a protein of 56, 178 D. Based on sequence homology, this ORF was confirmed to correspond to the nifA gene. Comparison of three nifA proteins (Klebsiella pneumoniae, Rhizobium meliloti, Rhizobium leguminosarum) revealed only a weak relationship in their N-terminal regions, whereas the C-terminal parts exhibited strong homology. Sequence analysis also showed that the R. leguminosarum nifA gene is followed by nifB and preceded by fixC with an open reading frame inserted in between. This novel ORF of 294 bp was found to be highly conserved also in R. meliloti. No known promoter and termination signals could be defined on the sequenced R. leguminosarum fragment. PMID- 3029678 TI - The nucleotide sequence of a 3.2 kb segment of mitochondrial maxicircle DNA from Crithidia fasciculata containing the gene for cytochrome oxidase subunit III, the N-terminal part of the apocytochrome b gene and a possible frameshift gene; further evidence for the use of unusual initiator triplets in trypanosome mitochondria. AB - A 3.2 kb segment of the maxicircle of Crithidia fasciculata mitochondrial (mt) DNA contains the gene for cytochrome oxidase subunit III (coxIII), the N-terminal portion of the gene for apocytochrome b (cytb) and two partially overlapping Unassigned Reading Frames (C.URF2/1). Transcript analysis of the segment reveals that both the coxIII gene and the C.URF2/1 area are transcribed into a pair of RNA products. With the C. fasciculata gene version as a probe, a coxIII gene could not be detected in nuclear and mtDNA of Trypanosoma brucei, indicating that the cytochrome oxidases of these two closely related trypanosome species may differ. The nucleotide homology in the N-terminal region of the coxIII and cytb genes in T. brucei, Leishmania tarentolae and C. fasciculata starts at a UUA leucine codon, which adds further support to the hypothesis that apart from AUG, other initiator triplets are used in trypanosomal mitochondria: UUG, CUG and UUA, all triplets coding for leucine in the universal code. Finally, the possibility is discussed that the two overlapping URFs (C.URF2/1) in fact represent a single, frameshift containing, gene. PMID- 3029680 TI - A dnaB-like protein of Pseudomonas aeruginosa. AB - A dnaB-like protein from P. aeruginosa was purified to near homogeneity using as an assay the immunoprecipitation by E. coli dnaB antiserum in a solid-phase. In the chromatographic characteristics including the affinity to immobilized ATP the dnaB-like protein of P. aeruginosa is similar to the dnaB protein of E. coli with the exception that it does not bind to heparin-Sepharose. The dnaB-like protein has a native molecular weight of about 320,000 as determined by glycerol gradient sedimentation. It consists of several identical subunits of molecular weight of 56,000 as measured in a denaturing SDS gel. Associated with the enzyme is a DNA dependent ATPase- and helicase activity. The dnaB-like protein is similar to the E. coli dnaB protein with regard to the binding of ATP gamma S and the formation of a ternary complex consisting of the enzyme, ATP gamma S, and phi X174 DNA. However, the enzyme of P. aeruginosa is inactive in a phi X174 DNA-dependent in vitro dnaB complementation assay using an E. coli dnaBts extract. PMID- 3029679 TI - Sequence homologies among the three yolk polypeptide (Yp) genes in Drosophila melanogaster. AB - To identify candidates for cis-acting sequences that regulate the sex-, stage-, and cell-specific expression of three coordinately regulated yolk polypeptide genes (Yp) in Drosophila melanogaster, we have mapped the Yp3 transcript, sequenced a 4278 bp DNA fragment containing the Yp3 gene, compared Yp3 region sequences to corresponding parts of Yp1 and Yp2, and compared the predicted amino acid sequence of YP3 to YP1 and YP2. The results showed that the Yps are largely homologous in translated regions, especially in the 3' half of the genes. Untranscribed flanking regions had little homology. A conserved inverted repeat (the H-box) has homology both to vertebrate steroid hormone receptor binding sites and to the ecdysone control region of Drosophila's hsp23. These results identify sequences to mutate in order to define elements that regulate Yp gene expression and govern YP polypeptide function. PMID- 3029681 TI - A new method for constructing linker scanning mutants. AB - A new procedure for the construction of linker scanning mutants is described. A plasmid containing the target DNA is randomly linearized and slightly shortened by a novel combination of established methods. After partial apurination with formic acid a specific nick or small gap is introduced at the apurinic site by exonuclease III, followed by nuclease S1 cleavage of the strand opposite the nick/gap. Synthetic linkers are ligated to the ends and plasmids having the linker inserted in the target DNA are enriched. Putative linker scanning mutants are identified by their topoisomer patterns after relaxation with topoisomerase I. This technique allows the distinction of plasmids differing in length by a single basepair. We have used this rapid and efficient strategy to generate a set of 32 linker scanning mutants covering the chicken lysozyme promoter from -208 to +15. PMID- 3029682 TI - The human ubiquitin gene family: structure of a gene and pseudogenes from the Ub B subfamily. AB - An ubiquitin cDNA clone was isolated from a human liver cDNA library. This clone contained two complete, and a portion of a third, ubiquitin coding sequences joined head to tail with no spacer peptides. Screening a human genomic library with a probe derived from the coding region of this cDNA identified a large number of cross-hybridising clones. Differential screening of these genomic clones with the 3' non-coding region of the cDNA identified three different 3' positive clones. Sequence analysis of these three clones revealed: a gene corresponding to the cDNA containing an intron in the 5' non-coding region and coding for three direct repeats of mature ubiquitin, and two related pseudogenes which appear to have arisen by reverse transcription and insertion into the genome. However, one pseudogene contains two repeats of the ubiquitin coding sequence, while the other contains only one. Hybridisation analysis of restricted human genomic DNA suggests the presence of one other closely related gene within the genome. PMID- 3029683 TI - Escherichia coli rep gene: sequence of the gene, the encoded helicase, and its homology with uvrD. AB - The sequence of a 2.67-kilobase section of the Escherichia coli chromosome that contains the rep gene has been determined. This gene codes for a protein of predicted Mr 72,800, a DNA helicase, which is also a single-stranded DNA dependent ATPase. The sequenced region contains an open reading frame of the correct length and orientation to encode the Rep protein. A secondary structure for the protein can be formulated from the amino acid sequence. We have compared both the primary and the secondary structures of Rep with other proteins and find the greatest homology between Rep and E. coli helicase II, the product of the uvrD gene. PMID- 3029684 TI - A DNA binding protein showing sequence specificity for a region containing the replication origin of Xenopus laevis mitochondrial DNA. AB - In Xenopus laevis mitochondria up to 14 different polypeptides with affinity for the DNA, have been identified by the protein blotting technique. Under stringent binding conditions only one polypeptide displayed specific affinity for a restriction fragment containing the H strand origin of replication of the Xenopus laevis mt chromosome. The proteins were fractionated by double stranded DNA cellulose chromatography. Under conditions which favor high affinity interactions between proteins and DNA, a protein of the 2M NaCl step shows specific binding to the DNA fragments containing the D-loop region. Some physical properties of the protein have been studied. It has a MW of 21.5 Kd and a globular shape as can be inferred from the relationship between MW and sedimentation coefficient (2.7 S). It binds non cooperatively to DNA and forms relatively stable complexes as demonstrated by DNA competition experiments. PMID- 3029685 TI - The complete coding sequence of the human A-raf-1 oncogene and transforming activity of a human A-raf carrying retrovirus. AB - The complete 606 amino acid sequence of the human A-raf oncogene has been deduced from the 2453 nucleotide sequence of a human T cell cDNA. A cysteine-rich region located near the amino terminus, which is highly conserved in A-raf and c-raf, shows significant homology with protein kinase C. A 5' deleted fragment of the cDNA has been incorporated into a murine retrovirus which endows the virus with the ability to transform cells in vivo and in vitro. Functionally, human A-raf is similar to v-raf and v-mos in that transformation is independent of ras gene function. PMID- 3029686 TI - Rice chloroplast DNA molecules are heterogeneous as revealed by DNA sequences of a cluster of genes. AB - We describe the isolation of two rice chloroplast HindIII fragments (9.5 kb and 5.3 kb) each containing a gene cluster coding for the large subunit of ribulose 1,5-bisphosphate carboxylase (rbcL), beta and epsilon subunits of ATPase (atpB and atpE), tRNAmet (trnM) and tRNAval (trnV). All five genes contained in the 9.5 kb fragment are potentially functional, whereas in the 5.3 kb fragment, rbcL is truncated and atpB is frame-shift mutated. The copy number of the 9.5 kb fragment is 10 times that of the 5.3 kb fragment, indicating that the two fragments are probably located on different chloroplast genomes and represent two different (major and minor) genomic populations. Thus, the rice chloroplast genome appears to be heterogeneous, contrary to general belief. We also describe the isolation of a rice mitochondrial HindIII fragment (6.9 kb) which contains an almost complete transferred copy of this chloroplast gene cluster. In this transferred copy, the coding sequences of rbcL, atpE and trnM contain perfectly normal reading frames, whereas atpB has become grossly defective and trnV is truncated. PMID- 3029687 TI - Polyadenylation at a cryptic site in the pBR322 portion of pSV2-neo: prevention of its utilization by the SV40 late poly(A) signal. AB - Transcripts originating from the SV40 late promoter of pSV2-neo or pSV2-cat contain pBR322 sequences and are polyadenylated at the SV40 late poly(A) site, resulting in an RNA of 3500 nt. If the SV40(L) poly(A) signal is destroyed, late orientation transcripts are polyadenylated at a site within pBR322 sequences, yielding in an RNA of 2500 nt. This cryptic poly(A) site is located 42-46 nucleotides downstream from an AAUAAA. Utilization of the pBR322 poly(A) signal is undetectable in late orientation transcripts from pSV2-neo or pSV2-cat, although it is located 966 nucleotides upstream from the SV40(L) poly(A) signal. The pBR322 site is not utilized when the spacing between the two poly(A) signals is varied from 209 to 1913 nucleotides. The pBR322 poly(A) site was utilized only in constructs in which all or portions of the SV40(L) poly(A) signal were deleted, such as in a construct with a 7 bp deletion into the SV40(L) AATAAA and adjacent sequences. PMID- 3029688 TI - Human, yeast and hybrid 3-phosphoglycerate kinase gene expression in yeast. AB - When the gene for yeast 3-phosphoglycerate kinase (PGK) is present on a high copy number plasmid in Saccharomyces cerevisiae, 30-40 percent of yeast protein is produced as PGK. However, when the structural part of this gene is replaced by as many as twenty different heterologous genes, production of gene products is greatly reduced--usually by more than 20 fold. This decrease in protein production is accompanied by large decreases in the steady-state levels of mRNA. However, in contrast to these coding sequences, replacement of the yeast PGK structural gene with a human PGK cDNA has little effect on the steady-state mRNA level in yeast. PGK is a two-domain enzyme and its 3-dimensional structure is highly conserved among species. These observations and others have led us to propose that the PGK protein itself might influence its own mRNA levels (Chen et al., Nucleic Acids Res. 12, pp. 8951-8969, 1984). In addition, data is presented here which suggest that the human PGK mRNA is less efficiently translated than the yeast PGK mRNA. Two different mechanisms of controlling gene expression are indicated. Both mechanisms appear to be independent of gene copy number. PMID- 3029690 TI - Structure and expression of an ethylene-related mRNA from tomato. AB - Messenger RNAs homologous to a cDNA clone (pTOM 13) derived from a ripe-tomato specific cDNA library are expressed during tomato fruit ripening and after the wounding of leaf and green fruit material. Both responses involve the synthesis of the hormone ethylene. Accumulation of the pTOM 13--homologous RNA during ripening is rapid and sustained, and reaches its maximum level in orange fruit. Following mechanical wounding of tomato leaves a pTOM 13--homologous RNA shows rapid induction within 30 minutes, which occurs before maximal ethylene evolution (2-3 h). This RNA also accumulates following the wounding of green tomato fruit. Northern blot analysis of poly(A)+ RNA indicates that the length of the mRNA is about 1400 nucleotides. Nucleotide sequence analysis showed the cDNA insert to contain the complete coding region of the pTOM 13 protein (33.5 kD) and an unusual 5' structure of ten dT-nucleotides. Hybridisation of the pTOM 13 cDNA insert to Southern blots of tomato DNA indicates the presence of only a small number of homologous sequences in the tomato genome. PMID- 3029689 TI - Cleavage of single stranded oligonucleotides by EcoRI restriction endonuclease. AB - The 31mer 5'-TCA ACG CTA GAA TTC GGA TCC ATC GCT TGG T, the complementary 33mer 5'-CCA AGC GAT GGA TCC GAA TTC TAG CGT TGA GAT, the 40mer 5'-GGC CAG GAT GGT GAA GAA TTC GAT CCG GTA CGT AGC TAA G, and the complementary 42mer 5'-TAC TTA GCT ACG TAC CGG ATC GAA TTC TTC ACC ATC CTG GCC were synthesized and their reactivity towards EcoRI was studied. It was found that the 31mer and the 40mer were cleaved at a comparable rate to the 31mer-33mer hybrid and the 40mer-42mer hybrid, respectively. The rate of cleavage of the 33mer and the 42mer was an order of magnitude lower. To rule out possible intermolecular duplex formation, the 33mer was immobilized on cellulose by ligation and labeled with alpha 32P-dCTP using Klenow fragment of E. coli DNA polymerase. EcoRI cleaved this immobilized oligomer into specific fragments. PMID- 3029691 TI - The chicken carbonic anhydrase II gene: evidence for a recent shift in intron position. AB - The complete nucleotide sequence of the coding region of the chicken carbonic anhydrase II (CA II) gene has been determined from clones isolated from a chicken genomic library. The sequence of a nearly full length chicken CA II cDNA clone has also been obtained. The gene is approximately 17 kilobase pairs (kb) in size and codes for a protein that is comprised of 259 amino acid residues. The 5' flanking region contains consensus sequences commonly associated with eucaryotic genes transcribed by RNA polymerase II. Six introns ranging in size from 0.3 to 10.2 kb interrupt the gene. The number of introns as well as five of the six intron locations are conserved between the chicken and mouse CA II genes. The site of the fourth intron is shifted by 14 base pairs further 3' in the chicken and thus falls between codons 147 and 148 rather than within codon 143 as in the mouse gene. Measurements of CA II RNA levels in various cell types suggest that CA II RNA increases in parallel with globin RNA during erythropoiesis and exists only at low levels, if at all, in non-erythroid cells. PMID- 3029692 TI - DNA sequence of the E. coli gyrB gene: application of a new sequencing strategy. AB - We have determined the sequence of the E. coli gyrB gene, using a new sequencing approach in which transposition from a mini-Mu plasmid into the DNA provides random start points for dideoxynucleotide sequence analysis. The gyrB sequence corresponds to a protein 804 amino acids long; a previously isolated protein fragment with partial enzymatic activity has been identified as the C-terminal half-molecule. A plausible terminator of gyrB transcription is located just beyond the structural gene. PMID- 3029693 TI - Improved Bordetella transformation. PMID- 3029694 TI - Cloning and characterization of the gene for Escherichia coli seryl-tRNA synthetase. AB - Seryl-tRNA synthetase is the gene product of the serS locus in Escherichia coli. Its gene has been cloned by complementation of a serS temperature sensitive mutant K28 with an E. coli gene bank DNA. The resulting clones overexpress seryl tRNA synthetase by a factor greater than 50 and more than 6% of the total cellular protein corresponds to the enzyme. The DNA sequence of the complete coding region and the 5'- and 3' untranslated regions was determined. Protein sequence comparison of SerRS with all available aminoacyl-tRNA synthetase sequences revealed some regions of significant homology particularly with the isoleucyl- and phenylalanyl-tRNA synthetases from E. coli. PMID- 3029696 TI - A new type of plasmid from a wild isolate of Dictyostelium species: the existence of closely situated long inverted repeats. AB - A circular plasmid having high copy number was found in a wild isolate of Dictyostelium species. Gel electrophoresis, electron microscopy and Southern blot hybridization revealed that the plasmid, named pDG1, is 4.5 Kb (1.5 micron) in size with closely situated long inverted repeats. The plasmid seems to be located in the nuclei. It was not a derivative of ribosomal DNA. The possible correlation of the plasmid with the putative intermediate DNA of retrotransposon DIRS-1 found in Dictyostelium discoideum is discussed. PMID- 3029695 TI - Expression and identification of immunity determinants on linear DNA killer plasmids pGKL1 and pGKL2 in Kluyveromyces lactis. AB - The linear dsDNA plasmids, pGKL1 (8.9 kb) and pGKL2 (13.4 kb) discovered in Kluyveromyces lactis, confer killer and immunity characteristics upon various yeast strains. We have devised an immunity assay and have been able to show the expression of an immunity phenotype in the K. lactis transformants harbouring conventional circular plasmids which contain DNA fragments of pGKL1. Using this expression system, the immunity determinant on pGKL1 was identified as ORF5. In addition, the presence of pGKL2 was proved to be essential for the expression of the immunity phenotype. This is the first demonstration of this new pGKL2 function, as distinct from its known functions for the replication and maintenance of pGKL1 in yeast cells. PMID- 3029697 TI - Structure of the gene for the stringent starvation protein of Escherichia coli. AB - The nucleotide sequence of the gene for the stringent starvation protein (SSP) of E. coli was determined. The deduced amino acid sequences shows that the SSP is composed of 212 amino acid residues, rich in both positively and negatively charged amino acids and has a molecular weight of 24,305. Primer extension experiments and nuclease S1 mapping analysis showed a site on the chromosome DNA corresponding to the 5' end of the transcript of the SSP gene. However, the consensus promoter sequences were not found at the upstream region. In the 3' flanking region a long coding frame was found immediately following the SSP gene, suggesting that the SSP gene is a member of a multicistronic operon. PMID- 3029698 TI - Shufflon: multi-inversion of four contiguous DNA segments of plasmid R64 creates seven different open reading frames. AB - The IncI alpha plasmid R64 was found to bear a highly mobile DNA segment which was designated as a clustered inversion region (J. Bacteriol. 165, 94-100, 1986). The clustered inversion region consists of four DNA segments designated respectively as A, B, C and D which differ in molecular size and restriction sites. The four DNA segments invert independently or in groups resulting in a complex DNA rearrangement. We now show the nucleotide sequence of the clustered inversion region of R64. The present results suggest that the clustered inversion region is a biological switch to select one of seven open reading frames whose primary structures at the region proximal to N-termini are constant while those at the C-terminal region are variable. A name, "Shufflon" was proposed to call this kind of the clustered inversion region. PMID- 3029699 TI - Overabundance of rare-cutting restriction endonuclease sites in the human genome. AB - A human chromosome 3-specific cosmid library was constructed from a somatic cell hybrid containing human chromosome 3 as its only human component. This library was screened to identify 230 human recombinants which contained an average insert size of 37 kilobases. DNA prepared from 54 of these cosmids, representing 2000 kilobases of human DNA, was then tested for restriction endonuclease sites for EcoRI, HindIII, KpnI, XhoI, and DraI, as well as those of the rare-cutting restriction endonucleases NotI, SfiI, NruI, MluI, SacII, and BssHII. Sites for the latter enzymes were much more abundant than would be expected from theoretical calculations, reflecting non-random clustering of these sites. This has important implications for the use of these enzymes in the construction of physical maps of chromosomes. Some individual cosmids contained large numbers of rare sites, offering an alternative means of physically mapping chromosomes based upon identifying clusters of rare restriction sites. These clusters appear to be spaced an average of 1000 kb apart. PMID- 3029700 TI - On the fidelity of DNA replication: herpes DNA polymerase and its associated exonuclease. AB - Procaryotic DNA polymerases contain an associated 3'----5' exonuclease activity which provides a proofreading function and contributes substantially to replication fidelity. DNA polymerases of the eucaryotic herpes-type viruses contain similar associated exonuclease activities. We have investigated the fidelity of polymerases purified from wild type herpes simplex virus, as well as from mutator and antimutator strains. On synthetic templates, the herpes enzymes show greater relative exonuclease activities, and greater ability to excise a terminal mismatched base, than procaryotic DNA polymerases which proofread. On a phi X174 natural DNA template, the herpes enzymes are more accurate than purified eucaryotic DNA polymerases; the error rate is similar to E. coli polymerase I. However, conditions which abnegate proofreading by E. coli polymerase I have little effect on the herpes enzymes. We conclude that either these viral polymerases are accurate in the absence of proofreading, or the conditions examined have little effect on proofreading by the herpes DNA polymerases. PMID- 3029701 TI - Structure of the Bombyx mori rDNA: initiation site for its transcription. AB - The initiation site of rDNA transcription in Bombyx mori was determined to be located at 909bp upstream from the 5'- end of mature 18s rRNA by S1-nuclease mapping and primer extension experiment. An in vitro transcription system, which was constructed using posterior silk glands of Bombyx larvae, initiated the transcription of cloned rDNA at exactly the same site as determined for the in vivo transcription above. The primary transcript seemed to be processed at about 200b downstream of the initiation site both in vivo and in vitro. Sequence analyses of the flanking region of the initiation site revealed that short repetitive sequences are widely distributed throughout the NTS region, and that a highly AT-rich region resides immediately upstream of the initiation region. PMID- 3029704 TI - myb-specific probes. PMID- 3029702 TI - Role in translation of a triple tandemly repeated sequence in the 5'-untranslated region of human thymidylate synthase mRNA. AB - A triple tandem repeat (TTR) consisting of 90 nucleotides exists immediately upstream of the ATG initiator codon in human thymidylate synthase (TS) cDNA (pcHTS-1). To investigate the role of the TTR in the expression of the TS cDNA, we used pcHTS-1 to construct mutant cDNA clones in which part of the TTR was deleted or an additional element was inserted. The mutant cDNA plasmid was introduced into murine TS-negative mutant cells and the relative translation efficiencies of the mutant cDNAs were determined by measuring the transient expression of TS activity and the amount of TS mRNA transcribed. The translation efficiency in transient expression of the mutants was increased by deletions covering all the first two repeated elements, and the part of the third closest to the ATG initiator codon, but was not affected by deletions of only parts of the first two repeated elements at the 5' end. The translation efficiency was also not affected by insertion of an additional repeated element into the TTR. These results suggest that the first two repeated elements at the 5' end both have inhibitory effects on translation of the TS mRNA, probably due to the unique structural feature of this element. PMID- 3029703 TI - Electroporation for the efficient transfection of mammalian cells with DNA. AB - A simple and reproducible procedure for the introduction of DNA into mammalian cells by electroporation is described. The parameters involving the cells, the DNA, and the electric field are investigated. The procedure has been applied to a broad range of animal cells. It is capable of transforming more than 1% of the viable cells to the stable expression of a selectable marker. PMID- 3029706 TI - Isolation and purification of CeqI endonuclease, an isoschizomer of EcoRV. PMID- 3029705 TI - Sequence of the Bacillus caldotenax and Bacillus stearothermophilus lctB genes. PMID- 3029707 TI - DNA inserted two bases down from the initiation site of a SP6 polymerase transcription vector is transcribed efficiently in vitro. PMID- 3029708 TI - Expression of active human blood clotting factor IX in transgenic mice: use of a cDNA with complete mRNA sequence. AB - Haemophilia B is a bleeding disorder caused by a functional deficiency of the clotting factor IX. A full length human factor IX complementary DNA clone containing all the natural mRNA sequences plus some flanking intron sequences was constructed with a metallothionein promoter and introduced into transgenic mice by microinjection into the pronuclei of fertilised eggs. The transgenic mice expressed high levels of messenger RNA, gamma-carboxylated and glycosylated protein, and biological clotting activity that are indistinguishable from normal human plasma factor IX. This study demonstrates the feasibility of expressing highly complex heterologous proteins in transgenic mice. It also provides the groundwork for the production of large amounts of human factor IX in larger transgenic livestock for therapeutic use, and the investigation of alternative genetic therapies for haemophilia B. PMID- 3029710 TI - Differential gene expression during the amoebal-plasmodial transition in Physarum. AB - We have prepared cDNA libraries for amoebae and plasmodia of the acellular slime mould, Physarum polycephalum. Differential screening was used to isolate cell type-specific cDNA clones (in bacteriophage M13) and both libraries yielded approximately 5% of such sequences. The amoebal- and plasmodial-specific clones were used to assay changes in transcription during the amoebal-plasmodial transition. The results obtained substantiate the view that the switch from amoebal to plasmodial characteristics occurs over several nuclear division cycles. With one exception, the specific cDNAs came from single-gene families. Southern blotting experiments also showed that they hybridised to identical restriction fragments from amoebal and plasmodial DNAs indicating that genomic rearrangements are unlikely to be involved in the regulation of these genes. PMID- 3029709 TI - The alpha protein ICP0 does not appear to play a major role in the regulation of herpes simplex virus gene expression during infection in tissue culture. AB - The herpes simplex virus type 1 (HSV-1) alpha protein ICP0 trans-activates HSV-1 early genes in transient expression assays. To investigate the function of ICP0 during HSV-1 infection, we have lowered the level of ICP0 by use of a recombinant plasmid that has been engineered to express the antisense message. Cell lines were constructed which stably carry the antisense plasmid. Total protein profiles from infected antisense cell lines showed that the level of ICP0 was reduced to less than 10% of the wild type level in two of the cell lines. However, reducing the level of ICP0 did not have a significant effect on the expression of HSV-1 early or late genes. The polypeptide patterns for the remaining infected cell polypeptides were similar in that no bands were absent although there were some quantitative differences. The level of two early proteins, glycoprotein B and glycoprotein D was reduced in one of the cell lines, however, levels were nearly equivalent to the control infection for two other cell lines tested. Virus yields were the same for the antisense cell lines and for parent cells. Decreased ICP0 levels did not lead to more restrictive phenotypes for an alpha 4 or alpha 27 mutant as protein patterns were similar for these mutants in antisense and parent cells. Therefore, while ICP0 has been demonstrated to be a strong inducer of gene expression in transient expression assays, it does not appear to have a major role as an activator during the productive infection of tissue culture cells. PMID- 3029711 TI - DNA orientation using specific avidin-ferritin biotin end labelling. AB - The orientation of DNA molecules has been determined by labelling one of the molecule end with a Biotin-labelled analog of dTTP (Bio-dUTP) and then by complexing the Bio-dUTP with Avidin-Ferritin. DNAs of phi X174, pBR322 and SV40 were end labelled with Bio-dUTP and imaged by Electron Microscopy (EM). This is a rapid, general method to unambiguously determine the orientation of DNA molecules for precise mapping and quantification of DNA secondary structures or protein-DNA interaction sites using EM. PMID- 3029712 TI - Recombination of DNAs in Xenopus oocytes based on short homologous overlaps. AB - Linear molecules of pBR322 and closely related plasmid DNAs were injected into Xenopus oocyte nuclei. Such molecules were degraded unless their ends were recombined. Non-homologous ends were joined rarely, if at all, but measurable recombination was supported by homologous sequences of less than 10 base pairs (bp). The efficiency of recombination increased as the length and degree of homology improved, in the range of about 8-20 bp. The homologous sequences had to be very close to the original molecular ends (within about 20 bp); internal homologies, even when they included better matches, were never used. These observations are best accommodated by a model of recombination which envisions exonucleolytic resection to expose homologous sequences, followed by annealing of single-stranded tails, tidying up and sealing of the new joint. Some of the recombined plasmids had novel tetracycline resistance genes; their properties give some insight into the function of the tet gene product. PMID- 3029713 TI - Progress towards construction of a total restriction fragment map of a human chromosome. AB - We present an approach to the construction of an overlapping restriction fragment map of a single human chromosome. A genomic cosmid library genome was constructed from a mouse-human hybrid cell line containing chromosome 17 as its only human genetic component. Cosmids containing human inserts were isolated by hybridisation to a human Alu sequence. DNAs from ninety-six randomly chosen cosmids were digested with either EcoRI or HindIII, end-labelled with 35S-dATP and analysed using agarose gel electrophoresis. Comparison of the restriction fragment patterns revealed two pairs of overlapping clones, that were confirmed by cross-hybridization of the overlapping fragments. The two pairs of cosmids both mapped to human chromosome 17, as shown by hybridization to a panel of somatic cell hybrids. These data demonstrate that the generation of an overlapping cosmid map along a human chromosome is feasible, representing an intermediate step towards the complete sequencing of a human chromosome. PMID- 3029714 TI - A rapid method to identify cosmids containing rare restriction sites. AB - A procedure for identifying specific cosmid clones containing recognition sites for "rare cutting" restriction enzymes has been developed. Cosmid clones containing human inserts were selected by hybridisation to human repetitive DNA. An oligonucleotide corresponding to the NotI recognition site, eight bases long, was labelled and used to probe DNA samples from one hundred cosmids. By optimising the difference in melting characteristics between eight-base perfect match and six-base match/two base mismatch hybrids, we were able to detect the cosmids containing either NotI (8 bp match) or XmaIII/EagI (6 bp match) sites. The generation of a map for rare cutter sites along a human chromosome, or a chromosome region, should be simplified using this approach, which will enable the identification of a set of "milestones" at intervals of several hundred kilobases (kb) along the DNA sequence. PMID- 3029715 TI - Conserved sequences near the early transcription start sites of vaccinia virus. AB - Transcription start sites were determined for the herpes simplex virus thymidine kinase (HSV-TK) mRNA expressed by four vaccinia virus recombinants in which the upstream insertion of shotgun-isolated vaccinia genomic fragments of 156 to 379 bp promoted this expression. Two of these fragments were related in such a manner that 62 bp separated two divergent early transcription start sites. The region of imperfect dyad symmetry revealed in this fragment is proposed to result from the presence of two divergent early transcription signals of vaccinia virus. Subsequent comparison showed that domains with high sequence homologies to those depicted by the dyad symmetry existed at comparable locations in the sequences flanking both the HSV-TK mRNA start site of the other two recombinants and that of several early vaccinia genes. Maximum homologies among these conserved sequences was obtained when they were aligned discontinuously. These studies also revealed a late mRNA start site with no more than 10 bp of vaccinia sequences upstream. PMID- 3029717 TI - Sequence and regulatory responses of a ribosomal protein gene from the fission yeast Schizosaccharomyces pombe. AB - We have determined the nucleotide sequence and mapped the 5' and 3' termini of a ribosomal protein gene. The gene is transcribed into a RNA molecule of about 770 nt and appears to initiate at multiple sites, as judged by SI nuclease analysis. Gene dosage experiments with a plasmid born gene leads to a proportional increase of the messenger RNA, but not to an overproduction of the protein, suggesting a posttranscriptional control mechanism. However, the heat shock response of this gene indicates that there is also a potential for transcriptional control. Comparison of the 5' flanking region of this gene with the ribosomal protein gene S 6 from Schizosaccharomyces pombe and with ribosomal protein genes from Saccharomyces cerevisiae revealed homologous sequences, which may be involved in the regulation of ribosomal protein genes. PMID- 3029716 TI - Tissue specific expression of the human alpha-1-antitrypsin gene in transgenic mice. AB - Normal and mutant human alpha-1-antitrypsin genes were cloned from a PiMZ heterozygous individual. Nucleotide sequence comparison demonstrated a T to C transition in exon III and an G to A transition in exon V of the PiZ gene. A 14.4 kb DNA fragment containing the entire PiM gene plus 2 kb of 5' and 3' flanking genomic DNA sequences was introduced into the germ line of mice and five F0 transgenic lines were established. Transgenic F1 progeny from F0 parents exhibited high levels of human alpha-1-antitrypsin protein in their plasma. The human gene was expressed primarily in liver of the transgenic mice as it is in man. However, expression of the human alpha-1-antitrypsin gene was also observed in kidneys of the transgenic mice, which led to the observation that the endogenous mouse gene is also expressed in the kidney. These data indicate that cis-acting elements within or proximal to the human alpha-1-antitrypsin gene are able to direct its in vivo transcription with a high degree of tissue specificity. PMID- 3029718 TI - Comparison of cauliflower mosaic virus 35S and nopaline synthase promoters in transgenic plants. AB - We have compared the level of expression of the Cauliflower Mosaic Virus 35S promoter and the nopaline synthase promoter when fused to a common reporter gene. A cassette containing the neomycin phosphotransferase (type II) coding sequence followed by the nopaline synthase 3' nontranslated region was used for transcriptional and translational evaluation of the two different promoters. These chimeric genes were introduced into petunia plants and the copy number of the gene, the steady state level of NPTII transcript and the levels of NPTII enzyme activity were determined. In this paper, we report that the NPT II transcript levels are on the average 30 fold higher in plants containing CaMV 35S promoter and leader sequences than in plants containing the same reporter gene but nopaline synthase promoter and leader sequences. Similarly, plants containing the CaMV 35S promoter had an average of 110 fold higher levels of NPTII enzyme activity than those containing the nopaline synthase promoter. The significance of these results for expression of foreign genes in plants is discussed. In addition, we describe the construction of a convenient plant expression cassette vector (pMON316) which utilizes the CaMV 35S promoter. PMID- 3029719 TI - Reactivity of modified ribose moieties of guanosine: new cleavage reactions mediated by the IVS of Tetrahymena precursor rRNA. AB - An RNA molecule consisting of the 5' exon and intervening sequence (IVS) of Tetrahymena precursor rRNA was oxidized with sodium periodate to convert the ribose moiety of the 3' terminal guanosine into a dialdehyde form. The modified RNA undergoes a specific cleavage reaction at the 5' splice site, but has no apparent cyclization activity. This novel reaction mediated by the IVS RNA is pH dependent over the range 6.5-8.5 and leaves a 5' phosphate and a 3'-OH at the newly created termini. The dialdehyde form of monomer guanosine is also capable of causing a specific cleavage reaction at the 5' splice site although the nucleotide is not covalently attached to the IVS RNA in the final product. These and other findings described in this report suggest that the cis diol of the intact ribose moiety of guanosine is not an absolute requirement for the IVS mediated reactions. PMID- 3029720 TI - pBR322 DNA inhibits simian virus 40 gene expression in Xenopus laevis oocytes. AB - SV40 DNA form I is expressed efficiently after its injection into the nuclei of Xenopus laevis oocytes, resulting in the synthesis of RNA and protein products of both viral late and early transcription units. However it was observed that injection of SV40 genes cloned into pBR322 or related plasmids yielded vastly reduced quantities of viral DNA and proteins. If SV40 DNA was cleaved from the plasmid, and then recircularized prior to microinjection, viral expression was regained. The inhibition by plasmid DNA was not confined to an effect in cis because coinjection of circular pBR322 DNA along with SV40 DNA, as separate entities, also blocked viral RNA and protein synthesis. As circular but not linear pBR322 DNA was actively transcribed by polymerase II in oocytes, even in the presence of SV40 DNA, it is likely that pBR322 competes for transcription factors required for viral gene expression. Injection of pBR322 as early as two hours after injection of SV40 DNA into the oocyte nucleus did not inhibit SV40 RNA synthesis, indicating that once initiated, SV40 transcription is stable and insensitive to the competition by plasmid DNA. A plasmid vector was developed that allows expression of SV40 DNA in Xenopus laevis oocytes. PMID- 3029721 TI - Regulation of viral and cellular promoter activity by polyomavirus early proteins. AB - The chloramphenicol-acetyl-transferase (CAT) expression system has been utilized to study the ability of the polyomavirus (Py) early proteins, the 100K large T, the 55K middle T and 22K small T-antigens, to activate a variety of eukaryotic promoters (the SV40 early, the alpha 2(1) collagen, the rabbit beta-globin, the polyomavirus early and the H-2 class I) in both transient and stable expression assays. We have found that either the complete polyomavirus early region or a plasmid expressing only the 55K middle T-antigen are capable of stimulating the expression of all the promoter-CAT plasmids in transient co-transfection experiments in both NIH-3T3 and Rat-2 cells. Conversely, the Py early proteins do not stimulate the transcription of most of the promoter-CAT genes stably introduced in the cell chromosomes, with the exception of H-2 class I promoter, when stimulation of transcription has been observed upon infection with recombinant retrovirus encoding the Py middle T-antigen. PMID- 3029722 TI - Characterization of the 5'-flanking region for the human fibrinogen beta gene. AB - To identify the possible regulatory sequences in the genetic expression of fibrinogen, a human genomic DNA library raised in lambda EMBL 4 phage was screened using cDNA probes coding for the A alpha, B beta and gamma chains of human fibrinogen. The entire fibrinogen locus was characterized and its organization analysed by means of hybridization and restriction mapping. Among the clones identified, a single recombinant lambda phage contained the beta gene and its 5'- and 3'-flanking regions. A 1.5 kb fragment of the immediate 5' flanking region was sequenced and S1 mapping experiments revealed three transcription start points. Comparison of this sequence with that previously reported for the same region upstream from the human gamma gene revealed no significant homology which suggests that the potential promoting sequences of these genes are different. In contrast, comparison of the 5'-flanking regions of human and rat beta genes revealed a 142 bp sequence of 80% homology situated 16 bp upstream from the human beta gene. This highly conserved region may well represents a potential candidate for a regulatory sequence of the human beta gene. PMID- 3029723 TI - Binding of anti-Z-DNA antibodies to negatively supercoiled SV40 DNA. AB - The binding of anti-Z-DNA antibody preparations to negatively supercoiled, protein-free SC40 DNA was analyzed. Covalent cross-linking with 0.1% glutaraldehyde followed by DNA restriction endonucleolytic fragmentation and nitrocellulose filtration allowed accurate mapping of antibody binding sites. The critical superhelical density necessary to allow antibody binding was -sigma = 0.056. The major region of antibody-DNA interaction was found within an SV40 segment spanning viral map positions 40 to 474. This region coincides with the nucleosome free region in SV40 minichromosomes and harbours the early and late promoter regions including the SV40 enhancer segment. Although it is unknown whether alternative, non-B-DNA conformations are generated in vivo within SV40 minichromosomes our results emphasize the high degree of DNA structural flexibility that can be realized under negative torsional stress. PMID- 3029724 TI - A nuclear protein with affinity for the 5' flanking region of a cell cycle dependent human H4 histone gene in vitro. AB - A nuclear protein with affinity for the 5' flanking region of a cell cycle dependent human H4 histone gene has been partially purified from nuclear extracts of human HeLa S3 cells. The region involved in the binding of the protein has been localized to an upstream DNA segment using an electrophoretic mobility shift assay. This DNA segment is devoid of RNA polymerase II consensus sequences and contains both homopurinic and A/T rich tracts. Analogous experiments have identified a similar, and perhaps identical, factor that has affinity for a cell cycle dependent human H3 histone gene promoter. This protein appears to bind to a DNA segment containing A/T rich sequences that bear homology with the binding region of the H4 histone promoter. Cell synchronization experiments have shown that the overall affinity of the protein(s) for the H3 and H4 histone 5' flanking regions in vitro is not dramatically altered during the cell cycle. Although the rate of histone gene transcription is modulated during early S phase, transcription occurs throughout the cell cycle. Hence, the protein(s) we have detected here may play a role in the basal expression of these genes. PMID- 3029725 TI - Telomeric site position heterogeneity in macronuclear DNA of Paramecium primaurelia. AB - In Paramecium primaurelia, the macronuclear gene encoding the G surface protein is located near a telomere. In this study, multiple copies of this telomere have been isolated and the subtelomeric and telomeric regions of some of them have been sequenced. The telomeric sequences consist of tandem repeats of the hexanucleotides C4A2 or C3A3. We show that the location where these repeats are added, which we call the telomeric site, is variable within a 0.6-0.8-kb region. These results are discussed in relation with the formation of macronuclear DNA. PMID- 3029726 TI - Nucleotide sequence of the pyruvate decarboxylase gene from Zymomonas mobilis. AB - Pyruvate decarboxylase (EC 4.1.1.1), the penultimate enzyme in the alcoholic fermentation pathway of Zymomonas mobilis, converts pyruvate to acetaldehyde and carbon dioxide. The complete nucleotide sequence of the structural gene encoding pyruvate decarboxylase from Zymomonas mobilis has been determined. The coding region is 1704 nucleotides long and encodes a polypeptide of 567 amino acids with a calculated subunit mass of 60,790 daltons. The amino acid sequence was confirmed by comparison with the amino acid sequence of a selection of tryptic fragments of the enzyme. The amino acid composition obtained from the nucleotide sequence is in good agreement with that obtained experimentally. PMID- 3029727 TI - The nucleotide sequence of the tnpA gene of Tn21. AB - The nucleotide sequence of the tnpA gene of Tn21 is presented. The transposase encoded by this gene is exactly the same length (988 amino acids) as the Tn501 transposase (4), and shows 72% homology overall with this protein, with greater homology towards the C-terminus. The sequence of the transposase is discussed in the context of the evolution of Class II transposable elements and of the characteristics of the enzyme's action. PMID- 3029728 TI - Nucleotide sequence of the streptomycinphosphotransferase and amidinotransferase genes from Streptomyces griseus. AB - Genes for streptomycin phosphotransferase and inosamine-P-amidinotransferase from a streptomycin-producing Streptomyces griseus were cloned on a 3.8kb BamHI-SphI fragment in S. lividans using the multicopy cloning vector pIJ702. The nucleotide sequence of this 3.8kb fragment was determined and the coding sequences for the two genes were identified by comparison with the amino-terminal sequences of the two enzymes purified from S. lividans clones. PMID- 3029730 TI - An efficient expression vector (pPLEX) for bacterial synthesis of non-fusion proteins and protein domains. PMID- 3029729 TI - Construction of the full coding information for murine J-chain protein from cDNA and its homologous genomic clone. PMID- 3029731 TI - [A case of familial multifocal chemodectoma]. PMID- 3029732 TI - Eicosanoids in skin UV inflammation--lack of leukotriene B4 elevation in UVB induced dermatitis. PMID- 3029733 TI - [Myocardial damage in children with rickets treated with vitamin D 3]. PMID- 3029735 TI - Comparison of neurothermography and contrast myelography. AB - Eighty consecutive patients who first had an electronic infrared thermogram (neurothermogram) and a subsequent oil myelogram were studied retrospectively. All patients had cervical and/or lumbosacral radiculopathy for 90 days. Negative neurothermograms were predictive of negative myelograms in 93% of our series. Positive neurothermograms were predictive of positive myelograms in 71% of the patients. The neurothermogram is not a specific test for intraspinal axis lesions as a cause of radiculopathy. Proximal neuropathic lesions of other causes will also produce abnormalities of the sympathetic autonomic (peripheral) nervous system. The major pathophysiologic findings detected by neurothermograms are complimentary to the EMG, NCVS, and late response techniques. Neurothermography was successful in predicting the outcome of myelography in 82% of patients in this study. PMID- 3029734 TI - [Cervico-facial metastasis of hepatocarcinoma. A case in Dakar]. PMID- 3029736 TI - Pathology of opportunistic infections in children with acquired immune deficiency syndrome. AB - Opportunistic infections (OI) were diagnosed by histology and culture of biopsy and autopsy material in 15 children with the acquired immune deficiency syndrome (AIDS). The opportunistic pathogens included Pneumocystic carinii, Toxoplasma gondii, Candida species, Aspergillus species, Mycobacterium avium-intracellulare, cytomegalovirus, and herpes simplex virus. Seven of 15 patients also had multiple systemic infections with common pathogenic bacteria. Accurate diagnosis of OI in AIDS is of importance in the decision regarding the choice of appropriate antimicrobial therapy. Careful histologic assessment of the biopsy specimens; awareness of unusual features such as paucity of organisms and inflammatory reaction, "histoid" variety of reaction, lack of granuloma formation, and resemblance to Whipple's disease in certain OI; and demonstration of causative organisms by appropriate special stains and/or culture are essential in the evaluation of these patients. PMID- 3029737 TI - ACTH-producing microadenoma of the pituitary in a young female infant with Cushing's disease: report of a case including immunocytologic and ultrastructural studies. AB - A female infant, younger than any other case in the literature, with ACTH producing microadenoma of the pituitary is reported. She had full-blown symptoms and signs as well as laboratory evidence of Cushing's disease. The tumor was investigated by histology, immunocytology (avidin-biotin-peroxidase complex technique), and electron microscopy. The possibility that the tumor was derived from the fetal intermediate lobe is discussed. PMID- 3029738 TI - Adrenoleukodystrophy: evidence for cytoplasmic inclusions in white blood cells. AB - Adrenoleukodystrophy (ALD) is a progressive neurological demyelinating disease associated with adrenal insufficiency. Pathognomonic lamellar profiles and clear clefts have been observed in several organs and tissues. We describe similar ultrastructural lamellar lipid profiles in white blood cells from 2 patients with ALD. It is suggested that electron microscopy of white blood cells may be a simple and effective screening method in patients presenting with a clinical picture suggestive of ALD. PMID- 3029739 TI - Congenital hybrid basal cell adenoma--adenoid cystic carcinoma of the salivary gland. AB - A congenital parotid gland tumor involving the facial nerve was excised, revealing histologic and ultrastructural features of a basal cell adenoma and adenoid cystic carcinoma. An asynchronous regional metastasis was successfully eradicated with adriamycin. Seven other similar appearing tumors occurring in neonates are documented in the literature and serve to establish this tumor as a clinicopathologic entity. The surrounding parotid gland had "dysplastic" changes attributed to obstruction during morphogenesis. PMID- 3029740 TI - Angiomatoid malignant fibrous histiocytoma with extensive lymphadenopathy simulating Castleman's disease. AB - We report the association in a 10-year-old boy of an angiomatoid malignant fibrous histiocytoma (AMFH) of the left thigh with ipsilateral inguinal, pelvic and extensive retroperitoneal lymphadenopathy, and severe systemic manifestations. These include growth retardation, fever, severe anemia, hypergammaglobinemia, and hypoalbuminemia. At ultrastructural level the tumor was characterized by an abundance of myofibroblasts, occasional histiocytes, and small vessels with marked reduplication of the basal lamina. Biopsies of the inguinal and abdominal lymph nodes showed follicular hyperplasia and massive plasmacytosis indistinguishable from Castleman's disease (giant lymph node hyperplasia) of plasma cell type. The radical surgical excision of the primary tumor in the thigh resulted in the disappearance of the abdominal lymphadenopathy and a marked reduction in size of the pelvic lymph nodes with marked decrease of the gammaglobulins, thus proving that the nodal lesions were the expression of a reactive process to the tumor rather than a coincidental independent lymphoproliferative disorder. Retroperitoneal and pelvic node dissection was performed 1 year after the radical excision of the thigh tumor because of persistent pelvic lymphadenopathy and failure of serum immunoglobulins M and A to return to normal level, with a recent peak of IgA to twofolds that of normal value. Metastatic AMFH was found in the three pelvic nodes. One month postoperatively IgA returned to near normal level whereas IgM remained slightly elevated. PMID- 3029741 TI - Identification of actin microfilaments in the intracytoplasmic inclusions present in recurring infantile digital fibromatosis (Reye tumor). AB - The diagnostic intracytoplasmic perinuclear inclusion bodies within the fibroblastic tumor cells of recurring digital fibrous tumor of childhood (Reye tumor) were found to be ultrastructurally composed of condensed microfilaments that were continuous with axially oriented cytoplasmic filament bundles running toward the cell membrane. Immunofluorescence microscopy performed with antibodies raised against vimentin and against actin revealed a strong positive reaction for vimentin in the tumor cell cytoplasm, whereas the spherical inclusion bodies were actin-positive. From these results the following conclusions may be drawn: (I) the mesenchymal origin of the Reye tumor cells is consistent with the identification of vimentin intermediate filaments and (II) the actin-positive spherical inclusion bodies appear to represent pathological aggregations of microfilaments. PMID- 3029742 TI - Prediction of metabolizable energy of broiler diets from chemical analysis. AB - Eighty-six broiler diets were studied to develop prediction equations for true metabolizable energy (TME) with nitrogen correction (TMEn) and calculated apparent metabolizable energy (AMEn) from chemical composition. Analyses were selected according to their capacity to represent the terms used in energy calculations, namely, dietary gross energy and energy excreted relative to diet consumed. Dietary gross energy was found to be highly correlated with ether extract alone. Prediction of excreta loss improved substantially with inclusion of two terms: a measurement of fiber [either crude fiber (CF), acid detergent fiber (ADF), or neutral detergent fiber (NDF)] and ash. A composite variable based on starch plus crude protein in place of fiber and ash yielded similar predictions. Accuracy of prediction of TMEn approached that of the biological analysis, but prediction of TME was somewhat less successful. Neutral detergent fiber was less satisfactory as a predictor than ADF or CF. Calculated AMEn was also highly predictable from ether extract or diet gross energy and a measurement of fiber alone; ash apparently was less helpful for AMEn. Previously published equations based on ether extract, ash, and an estimate of fiber gave results similar to the derived equations. PMID- 3029743 TI - [Post-crisis elevation of adrenocorticotropic hormone and prolactin in epileptic children]. AB - In six children, aged from 8 to 12 years, presenting with primary generalized epilepsy a significant rise in plasma ACTH and prolactin levels (as compared with levels measured 5 days later) was observed either during a generalized seizure demonstrated by EEG (3 cases) or within 1 hour of a generalized seizure (3 cases). The neurophysiological basis for these post-epileptic endocrine disturbances is a sudden depression of dopaminergic and GABA-ergic pathways. As a result, these endocrine changes, already well documented in adults, may be regarded as a good biological marker of the epileptic origin of a paroxysmal attack. PMID- 3029744 TI - [Acute renal failure caused by enalapril. 3 cases]. PMID- 3029745 TI - [Apparently idiopathic pes cavus in young adults: value of stimulo-detection electrodiagnosis. Preliminary results]. PMID- 3029746 TI - [Gastrointestinal infections in children. A strategy for searching pathogenic agents]. AB - From a bacteriological and virological study of 2056 stool samples obtained from 1595 children we have attempted to derive epidemiological arguments leading to an algorithm of coprodiagnosis. We believe that Rotavirus should be systematically searched for at any age, as it is the most frequent pathogen encountered (27% of children with positive detection). If the test is positive, no other test needs to be performed in patients staying less than 4 days in hospital. If the test is negative, or if the child's clinical condition requires prolonged hospitalization, conventional bacteriology must be carried out, including a search for Salmonella, Shigella and Yersinia. The search for Campylobacter should be limited to infections occurring in children under five. PMID- 3029747 TI - [Acute parvovirus B19 arthritis]. PMID- 3029748 TI - [Prostaglandin and cAMP levels in the endometrium of women with a normal menstrual cycle]. AB - The content of prostaglandins and cAMP was determined in the endometrium of healthy women with a regular menstrual cycle in its different phases. It was shown that the PGE2 level in the proliferative (the 7th-10th day of the cycle) and secretory (the 24th-26th day of the cycle) phases was the same. At the same time the level of PGE2 in the secretory phase of the cycle was 3 times higher than in the proliferative phase. The level of cAMP in the secretory phase of the cycle was 2 times higher than in the proliferative phase. PMID- 3029749 TI - [The role of the beta-adrenergic system in the function of calcium-regulating glands]. AB - The examination of 27 patients with coronary heart disease and experiments on 42 rabbits showed that intravenous administration of beta-adrenostimulant isoprenaline enhanced the blood level of parathormone and depressed that of calcitonin whereas administration of any beta-adrenoblocker showed contrary results. The patterns of adrenergic system effects on function of the calcium regulating glands were discussed. PMID- 3029750 TI - [Glucagonoma syndrome]. PMID- 3029751 TI - GABA synapses, depression, and antidepressant drugs. PMID- 3029752 TI - Antidepressant monoamine oxidase inhibitors enhance serotonin but not norepinephrine neurotransmission. PMID- 3029753 TI - Modulation of the benzodiazepine-GABA receptor chloride ionophore complex by multiple allosteric sites: evidence for a barbiturate "receptor". PMID- 3029754 TI - Beta-carbolines: new insights into the clinical pharmacology of benzodiazepine receptor ligands. PMID- 3029755 TI - Pharmacodynamic approaches to benzodiazepine action in man. PMID- 3029756 TI - Benzodiazepine-serotonin interactions in man. PMID- 3029757 TI - Regulation of beta adrenergic and serotonin-S2 receptors in brain by electroconvulsive shock and serotonin. PMID- 3029758 TI - Production of hepatitis B virus particles in Hep G2 cells transfected with cloned hepatitis B virus DNA. AB - The hepatoblastoma cell line Hep G2 was transfected with a plasmid carrying the gene that confers resistance to G418 and four 5'-3' tandem copies of the hepatitis B virus (HBV) genome positioned such that two dimers of the genomic DNA are 3'-3' with respect to one another. Cells of one clone that grew in the presence of G418 produce high levels of hepatitis B e antigen and of hepatitis B surface antigen. HBV DNA is carried by these cells as chromosomally integrated sequences and episomally as relaxed circular, covalently closed, and incomplete copies of the HBV genome. Viral DNA was detected also in conditioned growth medium at the buoyant densities characteristic for infectious Dane and immature core particles. Finally, HBV-specific components morphologically identical to the 22-nm spherical and filamentous hepatitis B surface antigen particles as well as 42-nm Dane particles were visualized by immunoelectron microscopic analysis. Therefore, we have demonstrated that the Hep G2 cell line can support the assembly and secretion not only of several of the replicative intermediates of HBV DNA but also of Dane-like particles. This in vitro system can now be used to study the life cycle of HBV and the reaction of immunocompetent cells with cells carrying HBV. PMID- 3029759 TI - Production of glycosylated physiologically "normal" human alpha 1-antitrypsin by mouse fibroblasts modified by insertion of a human alpha 1-antitrypsin cDNA using a retroviral vector. AB - Alpha 1-Antitrypsin (alpha 1AT) deficiency is a hereditary disorder characterized by reduced serum levels of alpha 1AT, resulting in destruction of the lower respiratory tract by neutrophil elastase. As an approach to augment alpha 1AT levels in this disorder with physiologically normal human alpha 1AT, we have integrated a full-length normal human alpha 1AT cDNA into the genome of mouse fibroblasts. To accomplish this, the retroviral vector N2 was modified by inserting the simian virus 40 early promoter followed by the alpha 1AT cDNA. Southern analysis demonstrated that the intact cDNA was present in the genome of selected clones of the transfected murine fibroblasts psi 2 and infected NIH 3T3. The clones produced three mRNA transcripts (5.8, 4.8, and 2.4 kilobases) containing human alpha 1AT sequences, secreted an alpha 1AT molecule recognized by an anti-human alpha 1AT antibody, with the same molecular mass (52 kDa) as normal human alpha 1AT and that complexed with and inhibited human neutrophil elastase. The psi 2 produced alpha 1AT was glycosylated, and when infused intravenously into mice, it had a serum half-life similar to normal alpha 1AT purified from human plasma and markedly longer than that of nonglycosylated human alpha 1AT cDNA-directed yeast-produced alpha 1AT. These studies demonstrate the feasibility of using a retroviral vector to insert the normal human alpha 1AT cDNA into non-alpha 1AT-producing cells, resulting in the synthesis and secretion of physiologically "normal" human alpha 1AT. PMID- 3029760 TI - Papillomavirus sequences integrate near cellular oncogenes in some cervical carcinomas. AB - The chromosomal locations of cellular sequences flanking integrated papillomavirus DNA in four cervical carcinoma cell lines and a primary cervical carcinoma have been determined. The two human papillomavirus (HPV) 16 flanking sequences derived from the tumor were localized to chromosome regions 20pter--- 20q13 and 3p25----3qter, regions that also contain the protooncogenes c-src-1 and c-raf-1, respectively. The HPV 16 integration site in the SiHa cervical carcinoma derived cell line is in chromosome region 13q14----13q32. The HPV 18 integration site in SW756 cervical carcinoma cells is in chromosome 12 but is not closely linked to the Ki-ras2 gene. Finally, in two cervical carcinoma cell lines, HeLa and C4-I, HPV 18 DNA is integrated in chromosome 8, 5' of the c-myc gene. The HeLa HPV 18 integration site is within 40 kilobases 5' of the c-myc gene, inside the HL60 amplification unit surrounding and including the c-myc gene. Additionally, steady-state levels of c-myc mRNA are elevated in HeLa and C4-I cells relative to other cervical carcinoma cell lines. Thus, in at least some genital tumors, cis-activation of cellular oncogenes by HPV may be involved in malignant transformation of cervical cells. PMID- 3029761 TI - Insulin-like growth factor II in human adrenal pheochromocytomas and Wilms tumors: expression at the mRNA and protein level. AB - Two forms of insulin-like growth factor (IGF) II with molecular masses of 10 and 7.5 kDa, respectively, were found in tumor tissue from human adrenal pheochromocytomas. The tumors contained 5.3-7.1 micrograms of immunoreactive IGF II per g of tissue, which is about 20 times more than in adrenal medulla. The total bioactive IGF in the pheochromocytomas exceeded that in normal liver or kidney, which contained only the 7.5-kDa IGF-II species, by a factor of approximately equal to 100. By contrast, the amount of IGF-I was just measurable and did not vary significantly between tumor and normal tissue. The high amounts of IGF-II in the pheochromocytomas were not reflected, however, by a corresponding increase of mRNA. The opposite situation was found in Wilms tumors, where IGF-II content was in the same range as in nontumor tissues despite increased expression of IGF-II mRNA. PMID- 3029763 TI - Specificity and efficiency of editing of mismatches involved in the formation of base-substitution mutations by the 3'----5' exonuclease activity of phage T4 DNA polymerase. AB - The specificity and efficiency of base-mispair editing by the 3'----5' exonuclease activity of phage T4 DNA polymerase has been measured using a sensitive infectivity assay. A series of oligodeoxynucleotide primer chains was synthesized chemically. These primers, when hybridized to phi X174 single stranded DNAs containing an amber codon, result in a mispaired nucleotide at the 3'-hydroxyl end of the primer chain within the amber codon. DNA synthesis on these primer X templates without the removal of the mispaired terminal nucleotide results in the formation of heteroduplex molecules that yield viable revertants upon transfection into an amber nonsuppressor host. This method permits determination of the efficiency of editing of a mismatch to 1 in 10(6) mismatches that escape editing and allows all eight mispairs that can yield viable revertants at an amber codon to be studied. The results of experiments with primers hybridizing to phi X174 am16 and am3 codons show that the order of mispair editing by T4 DNA polymerase is Ttemplate X Gprimer less than (A X G, T X C) less than (T X T, G X A, G X G, A X C) less than A X A. The efficiency of editing depends upon the mispair, as well as the neighboring DNA sequence. Under these conditions of synthesis, the 3'----5' exonuclease activity, depending upon the mispair and DNA sequences beyond the nearest neighbors, is estimated to contribute a factor of from 2.3 X 10(3)- to greater than 10(6)-fold to the accuracy of T4 DNA polymerase. PMID- 3029762 TI - Differential localization of calmodulin-dependent enzymes in rat brain: evidence for selective expression of cyclic nucleotide phosphodiesterase in specific neurons. AB - High-affinity antibodies against calmodulin (CaM)-dependent cyclic nucleotide phosphodiesterase and protein phosphatase (calcineurin) were purified and characterized. Rabbit anti-phosphodiesterase antibody did not react with other phosphodiesterases or with the regulatory subunits of cAMP-dependent protein kinase. Affinity-purified goat anti-calcineurin antibody recognized both the 61 kDa catalytic subunit and the 18-kDa Ca2+-binding subunit of the phosphatase. Neither antibody reacted with CaM, several CaM-binding proteins (calmodulin dependent protein kinase, myosin light chain kinase, fodrin), or other cytosolic proteins from brain. The antibodies were used to compare the cellular localization of these two CaM-dependent enzymes in rat brain. Both calcineurin and phosphodiesterase were found predominantly in nerve cells; however, phosphodiesterase was restricted to very specific neuronal populations. Phosphodiesterase was prominent in the somatic cytoplasm and dendrites of regional output neurons--e.g., cerebellar Purkinje cells and hippocampal and cortical pyramidal cells. The extensive and uniform staining in the dendrites was consistent with postsynaptic localization and suggested an important function for this enzyme in neurons that integrate multiple convergent inputs. Calcineurin was present in virtually all classes of neurons, with immunoreactivity confined primarily to cell bodies. Both diffuse cytoplasmic staining and characteristic punctate staining of cell bodies were observed; the latter suggested compartmentalization of calcineurin at or near the plasma membrane. The results of this study demonstrate that calcineurin and phosphodiesterase are differentially localized in the central nervous system. Thus, the expression and compartmentalization of CaM-binding proteins may be highly regulated and specific for particular differentiated nerve cell types. PMID- 3029764 TI - II-BGlc, a glucose receptor of the bacterial phosphotransferase system: molecular cloning of ptsG and purification of the receptor from an overproducing strain of Escherichia coli. AB - The bacterial phosphoenolpyruvate:glycose phosphotransferase system (PTS) consists of interacting cytoplasmic and membrane proteins that catalyze the phosphorylation and translocation of sugar substrates across the cell membrane. One PTS protein, II-BGlc, is the membrane receptor specific for glucose and methyl D-glucopyranosides; the protein has been purified to homogeneity from Salmonella typhimurium [Erni, B., Trachsel, H., Postma, P. & Rosenbusch, J. (1982) J. Biol. Chem. 257, 13726-13730]. In the present experiments, the Escherichia coli ptsG locus, which encodes II-BGlc, was isolated from a transducing phage library and subcloned into plasmid vectors. The resulting plasmids complement the following phenotypic defects of ptsG mutants: growth on glucose, uptake and phosphorylation of methyl alpha-D-glucoside, and repression of the utilization of non-PTS sugars by methyl alpha-glucoside. The transformed cells overproduce II-BGlc 4- to 10-fold, and a Mr 43,000 polypeptide was synthesized from the plasmids in an in vitro transcription/translation system. The E. coli and S. typhimurium II-BGlc proteins differ in their physical properties, and a modified, three-step purification procedure was developed for isolating the E. coli protein. PMID- 3029766 TI - Lateral diffusion of lipids in membranes by pulse saturation recovery electron spin resonance. AB - Short-pulse saturation recovery electron spin resonance methods have been used to measure lateral diffusion of nitroxide-labeled lipids in multilamellar liposomal dispersions. Nitroxides with 14N and 15N isotopes introduced both separately and together were used. Differential equations have been written and solved for complex saturation recovery signals involving several superimposed exponentials. The time constants contain various combinations of the spin-lattice relaxation time (T1e) for both isotopes, Heisenberg exchange rates, and nuclear spin-lattice relaxation times (T1n). Signals of high quality were fitted by Monte Carlo variation of the amplitudes and time constants. The reliability of the approach was tested extensively by verifying that (i) the predicted number of exponentials agreed with the experimental number, (ii) relaxation parameters that were determined were independent of the observed hyperfine transition, (iii) the time constants were independent of saturating pulse length, (iv) T1e and T1n do not change when Heisenberg exchange is changed by varying the concentration, and (v) Heisenberg exchange is indeed proportional to the concentration. It has been established that bimolecular collision rates over a wide range of conditions can be reliably measured using the methodology described here. The methods depend on the favorable match of bimolecular collision rates at micromolar concentrations to nitroxide spin-lattice relaxation probabilities. PMID- 3029765 TI - Roles of DNA topoisomerases in simian virus 40 DNA replication in vitro. AB - We examined the roles of DNA topoisomerases in the replication of simian virus 40 (SV40) DNA in a cell-free system composed of an extract from HeLa cells supplemented with purified SV40 tumor antigen. When the activities of both topoisomerase I (EC 5.99.1.2) and topoisomerase II (EC 5.99.1.3) in the extract were blocked with specific inhibitors or antibodies, DNA synthesis was decreased by a factor of 15-20. Addition of purified HeLa DNA topoisomerase II to extracts immunologically depleted of both topoisomerases completely restored replication, and the replication products consisted largely of monomeric daughter molecules. Addition of purified HeLa DNA topoisomerase I to depleted extracts restored DNA synthesis, but the primary products were multiply intertwined, catenated daughter molecules. We conclude that DNA topoisomerases have at least two roles in the replication of SV40 DNA. Either topoisomerase I or topoisomerase II is sufficient to provide the unlinking activity necessary for fork propagation during SV40 DNA replication. However, topoisomerase II is uniquely required for the segregation of newly synthesized daughter molecules. PMID- 3029767 TI - Developmental acquisition of DNase I sensitivity of the phosphoenolpyruvate carboxykinase (GTP) gene in rat liver. AB - The sensitivity to DNase I digestion of the gene encoding rat phosphoenolpyruvate carboxykinase (GTP) (EC 4.1.1.32) was assessed during development and prior to the onset of expression. This gene is resistant to DNase I digestion in nuclei isolated from livers of 19-day rat fetuses. Gradual acquisition of sensitivity of the phosphoenolpyruvate carboxykinase gene, which starts later than the 19th day of gestation and is completed by the 21st day, occurs before initiation of gene expression. As transcription of the phosphoenolpyruvate carboxykinase gene is not detected until birth, the events observed may represent a shift from a dormant to an active gene. Injection of N6,O2-dibutyryladenosine 3',5'-cyclic monophosphate into fetuses on the 19th day of gestation induces gene expression and sensitivity to DNase I digestion within 3 hr of treatment. While this short treatment does not affect the methylation pattern of the gene, longer treatment of fetuses (2 days) with dibutyryl-cAMP results in premature hypomethylation of the gene. A hierarchy of modifications of the phosphoenolpyruvate carboxykinase gene during development is discussed. PMID- 3029769 TI - A promoterless retroviral vector indicates that there are sequences in U3 required for 3' RNA processing. AB - We constructed a retrovirus vector in which all viral transcriptional signals are deleted during provirus formation. This vector would be safer and more useful in gene-transfer experiments. Construction of this vector involved the deletion of all of the U3 sequence except for 10 base pairs, required for integration, from the right-side long terminal repeat. We found that this deletion resulted in an inability to propagate virus efficiently. When we inserted sequences containing all of the signals for polyadenylylation of simian virus 40 late mRNA at the end of our vector, we were able to propagate virus efficiently. This result indicates that there are sequences in U3 upstream from AAUAAA that are essential for 3' processing of viral RNA. PMID- 3029768 TI - Chromatin structure is required to block transcription of the methylated herpes simplex virus thymidine kinase gene. AB - Inhibition of herpes simplex virus (HSV) thymidine kinase (TK) gene transcription (pHSV-106, pML-BPV-TK4) by DNA methylation is an indirect effect, which occurs with a latency period of approximately equal to 8 hr after microinjection of the DNA into TK- rat 2 and mouse LTK- cells. We have strong evidence that chromatin formation is critical for the transition of the injected DNA from methylation insensitivity to methylation sensitivity. Chromatin was reconstituted in vitro by using methylated and mock-methylated HSV TK DNA and purified chicken histone octamers. After microinjection, the methylated chromatin was always biologically inactive, as tested by autoradiography of the cells after incubation with [3H]thymidine and by RNA dot blot analysis. However, in transformed cell lines, reactivation of the methylated chromatin occurred after treatment with 5 azacytidine. Furthermore, integration of the TK chromatin into the host genome is not required to block expression of the methylated TK gene. Mouse cells that contained the pML-BPV-TK4 chromatin permanently in an episomal state also did not support TK gene expression as long as the TK DNA remained methylated. PMID- 3029770 TI - Regulation of c-myc and c-fos mRNA levels by polyomavirus: distinct roles for the capsid protein VP1 and the viral early proteins. AB - The levels of c-myc, c-fos, and JE mRNAs accumulate in a biphasic pattern following infection of quiescent BALB/c 3T3 mouse cells with polyomavirus. Maximal levels of c-myc and c-fos mRNAs were seen within 1 hr and were nearly undetectable at 6 hr after infection. At 12 hr after infection mRNA levels were again maximal and remained elevated thereafter. Empty virions (capsids) and recombinant VP1 protein, purified from Escherichia coli, induced the early but not the late phase of mRNA accumulation. Virions, capsids, and recombinant VP1 protein stimulated [3H]thymidine nuclear labeling and c-myc mRNA accumulation in a dose-responsive manner paralleling their affinity for the cell receptor for polyoma. The second phase of mRNA accumulation is regulated by the viral early gene products, as shown by polyomavirus early gene mutants and by a transfected cell line (336a) expressing middle tumor antigen upon glucocorticoid addition. These results suggest that polyomavirus interacts with the cell membrane at the onset of infection to increase the levels of mRNA for cellular genes associated with cell competence for DNA replication, and subsequently these levels are maintained by the action of the early viral proteins. PMID- 3029771 TI - The E2 "gene" of bovine papillomavirus encodes an enhancer-binding protein. AB - The E2 early open reading frame (presumably gene) of bovine papillomavirus-1 was fused in frame with the collagen-beta-galactosidase-encoding region of the vector pJG200 and was expressed in and partially purified from Escherichia coli. The hybrid protein specifically bound to the enhancer region of bovine papillomavirus at several sites. DNase I-cleavage protection analysis of one such site revealed the protected sequence. A comparison of the protected sequence with the remainder of the DNA sequences that also have affinity for the protein revealed a consensus sequence having the motif AATTGGCGGNNCG, in which N is any nucleotide. The protected region also includes a sequence with 2-fold rotational symmetry- ATCGGTG/CACCGAT. PMID- 3029772 TI - Cloning and expression of cDNA for human poly(ADP-ribose) polymerase. AB - cDNAs encoding poly(ADP-ribose) polymerase from a human hepatoma lambda gt11 cDNA library were isolated by immunological screening. One insert of 1.3 kilobases (kb) consistently hybridized on RNA gel blots to an mRNA species of 3.6-3.7 kb, which is consistent with the size of RNA necessary to code for the polymerase protein (116 kDa). This insert was subsequently used in both in vitro hybrid selection and hybrid-arrested translation studies. An mRNA species from HeLa cells of 3.6-3.7 kb was selected that was translated into a 116-kDa protein, which was selectively immunoprecipitated with anti-poly (ADP-ribose) polymerase. To confirm that the 1.3-kb insert from lambda gt11 encodes for poly(ADP-ribose) polymerase, the insert was used to screen a 3- to 4-kb subset of a transformed human fibroblast cDNA library in the Okayama-Berg vector. One of these vectors [pcD-p(ADPR)P; 3.6 kb] was tested in transient transfection experiments in COS cells. This cDNA insert contained the complete coding sequence for polymerase as indicated by the following criteria: A 3-fold increase in in vitro activity was noted in extracts from transfected cells compared to mock or pSV2-CAT transfected cells. A 6-fold increase in polymerase activity in pcD-p(ADPR)P transfected cell extracts compared to controls was observed by "activity gel" analysis on gels of electrophoretically separated proteins at 116 kDa. A 10- to 15-fold increase in newly synthesized polymerase was detected by immunoprecipitation of labeled transfected cell extracts. Using pcD-p(ADPR)P as probe, it was observed that the level of poly(ADP-ribose) polymerase mRNA was elevated at 5 and 7 hr of S phase of the HeLa cell cycle, but was unaltered when artificial DNA strand breaks are introduced in HeLa cells by alkylating agents. PMID- 3029773 TI - Proton NMR measurements of bacteriophage T4 lysozyme aided by 15N isotopic labeling: structural and dynamic studies of larger proteins. AB - A strategy for resolution and assignment of single proton resonances in proteins of molecular mass up to at least 40 kDa is presented. This approach is based on 15N (or 13C) labeling of selected residues in a protein. The resonances from protons directly bonded to labeled atoms are detected in a two-dimensional 1H-15N (or 13C) spectrum. The nuclear Overhauser effects from isotopically tagged protons are selectively observed in one-dimensional isotope-directed measurements. Using this approach, we have observed approximately 160 resonances from 15N-bonded protons in the backbone and sidechains of uniformly 15N-labeled T4 lysozyme (molecular mass = 18.7 kDa). Partial proton-deuterium exchange can be used to simplify the 1H-15N spectrum of this protein. These resonances are identified by amino acid class using selective incorporation of 15N-labeled amino acids and are assigned to specific residues by mutational substitution, multiple 15N and 13C labeling, and isotope-directed nuclear Overhauser effect measurements. For example, using a phenyl[15N]alanine-labeled lysozyme variant containing two consecutive phenylalanine residues in an alpha-helical region, we observe an isotope-directed nuclear Overhauser effect from the amide proton of Phe-66 to that of Phe-67. PMID- 3029774 TI - Yeast cytochrome c with phenylalanine or tyrosine at position 87 transfers electrons to (zinc cytochrome c peroxidase)+ at a rate ten thousand times that of the serine-87 or glycine-87 variants. AB - Of the many factors known to influence the rate of electron transfer between two metalloproteins, it is particularly difficult to assess the role of the polypeptide matrix intervening between the donor and acceptor sites. To determine whether the phylogenetically conserved Phe-87 of yeast iso-1-cytochrome c helps to mediate electron transfer between cytochrome c and cytochrome c peroxidase, we have constructed mutants of cytochrome c that are altered at this position and now have studied the kinetics of long-range electron transfer within their complexes with zinc-substituted cytochrome c peroxidase. We find that the rate of electron transfer from reduced cytochrome c to the zinc cytochrome c peroxidase pi-cation radical is four orders of magnitude greater when phenylalanine or tyrosine is present at position 87 than when serine or glycine is present. PMID- 3029776 TI - Inducible binding of a factor to the c-fos regulatory region. AB - The c-fos gene is rapidly and transiently activated in quiescent BALB/c-3T3 cells in response to serum, platelet-derived growth factor or conditioned medium from v sis-transformed cells. This activation occurs at the level of transcription and in the absence of new protein synthesis. Using a gel electrophoresis DNA-binding assay, we have found a DNA-binding activity in BALB/c-3T3 cells that is induced within 20 min of treatment with conditioned medium from v-sis-transformed cells. A DNA methylation interference assay has shown that this factor binds to a sequence approximately 346 base pairs upstream of the transcription initiation site of the human c-fos gene. Insulin, epidermal growth factor, and phorbol 12 myristate 13-acetate fail to induce this DNA-binding factor. Protein synthesis inhibitors do not block the induction of this activity. We propose that this factor preexists in an inactive form in quiescent cells and that its binding activity is activated in response to appropriate extracellular inducers. PMID- 3029775 TI - Purification of the colony-stimulating factor 1 receptor and demonstration of its tyrosine kinase activity. AB - Colony-stimulating factor 1 (CSF-1) regulates the survival, proliferation, and differentiation of mononuclear phagocytes. The CSF-1 receptor was purified from cell membranes of the J774.2 mouse macrophage cell line by solubilization with Triton X-100, CSF-1 affinity chromatography, and gel filtration. The purified receptor is a protein or glycoprotein of 165 kDa comprising a single polypeptide chain that is not covalently associated, either as a homopolymer, or with any other protein. CSF-1 stimulated autophosphorylation of the purified receptor in tyrosine residues. Casein but not histone was shown to act as a substrate for the tyrosine protein kinase activity of purified receptor. PMID- 3029777 TI - Cell-type-specific responses of RT4 neural cell lines to dibutyryl-cAMP: branch determination versus maturation. AB - This report describes the induction of cell-type-specific maturation, by dibutyryl-cAMP and testololactone, of neuronal and glial properties in a family of cell lines derived from a rat peripheral neurotumor, RT4. This maturation allows further understanding of the process of determination because of the close lineage relationship between the cell types of the RT4 family. The RT4 family is characterized by the spontaneous conversion of one of the cell types, RT4-AC (stem-cell type), to any of three derivative cell types, RT4-B, RT4-D, or RT4-E, with a frequency of about 10(-5). The RT4-AC cells express some properties characteristic of both neuronal and glial cells. Of these neural properties expressed by RT4-AC cells, only the neuronal properties are expressed by the RT4 B and RT4-E cells, and only the glial properties are expressed by the RT4-D cells. This in vitro cell-type conversion of RT4-AC to three derivative cell types is a branch point for the coordinate regulation of several properties and seems to resemble determination in vivo. In our standard culture conditions, several other neuronal and glial properties are not expressed by these cell types. However, addition of dibutyryl-cAMP induces expression of additional properties, in a cell-type-specific manner: formation of long cellular processes in the RT4-B8 and RT4-E5 cell lines and expression of high-affinity uptake of gamma-aminobutyric acid, by a glial-cell-specific mechanism, in the RT4-D6-2 cell line. These new properties are maximally expressed 2-3 days after addition of dibutyryl-cAMP. This indicates that conversion of RT4-AC to the derivative cell types is also a branch point for the regulation of cell-type-specific properties whose expression is responsive to cAMP. Thus, the potential for maturation in response to increased cAMP is a property that segregates in a cell-type-specific manner and is activated at the determinational level in this system. PMID- 3029778 TI - Transfection of a rearranged viral DNA fragment, WZhet, stably converts latent Epstein-Barr viral infection to productive infection in lymphoid cells. AB - An Epstein-Barr viral gene (ZEBRA) is identified that, in human lymphoblastoid cells, activates a switch causing the virus to shift from the latent to the replicative phase of its life cycle. We have shown that a 2.7-kilobase-pair rearranged Epstein-Barr virus DNA fragment of this gene (BamHI fragment WZhet) induced transient expression of viral replicative antigens and polypeptides when it was transfected into a somatic cell hybrid, which was derived from the fusion of an epithelial line cell with a Burkitt lymphoma cell. We now show that this rearranged WZhet fragment, when introduced stably into lymphoblastoid cells, will activate expression of the complete viral replicative cycle in 1-10% of the lymphoblastoid cells, leading to production of biologically active virions that can immortalize primary lymphocytes. The transfected plasmid appears to be regulated in a manner analogous to the complete Epstein-Barr virus genome. PMID- 3029779 TI - Direct measurement of free radical generation following reperfusion of ischemic myocardium. AB - Electron paramagnetic resonance spectroscopy was used to directly measure free radical generation in perfused rabbit hearts. Hearts were freeze-clamped at 77 degrees K during control perfusion, after 10 min of normothermic global ischemia (no coronary flow), or following post-ischemic reperfusion with oxygenated perfusate. The spectra of these hearts exhibited three different signals with different power saturation and temperature stability: signal A was isotropic with g = 2.004; signal B was anisotropic with axial symmetry with g parallel = 2.033 and g perpendicular = 2.005; signal C was an isotropic triplet with g = 2.000 and hyperfine splitting an = 24 G (1 G = 0.1 mT). The g values, linewidth, power saturation, and temperature stability of signal A are identical to those of a carbon-centered semiquinone, whereas those of signal B are similar to alkyl peroxyl or superoxide oxygen-centered free radicals; signal C is most likely a nitrogen-centered free radical. In the control heart samples signal A predominated, whereas in ischemic hearts signal A decreased in intensity, and signals B and C became more intense; with reperfusion all three signals markedly increased. Free radical concentrations derived from the intensities of the B and C signals peaked 10 sec after initiation of reflow. At this time the oxygen centered free radical concentration derived from the intensity of signal B was increased over six times the concentration measured in control hearts and over two times the concentration measured in ischemic hearts. Hypoxic reperfusion did not increase any of the free radical signals over the levels observed during ischemia. These experiments directly demonstrate that reactive oxygen-centered free radicals are generated in hearts during ischemia and that a burst of oxygen radical generation occurs within moments of reperfusion. PMID- 3029780 TI - Basal and adenosine receptor-stimulated levels of cAMP are reduced in lymphocytes from alcoholic patients. AB - Alcoholism causes serious neurologic disease that may be due, in part, to the ability of ethanol to interact with neural cell membranes and change neuronal function. Adenosine receptors are membrane-bound proteins that appear to mediate some of the effects of ethanol in the brain. Human lymphocytes also have adenosine receptors, and their activation causes increases in cAMP levels. To test the hypothesis that basal and adenosine receptor-stimulated cAMP levels in lymphocytes might be abnormal in alcoholism, we studied lymphocytes from 10 alcoholic subjects, 10 age- and sex-matched normal individuals, and 10 patients with nonalcoholic liver disease. Basal and adenosine receptor-stimulated cAMP levels were reduced 75% in lymphocytes from alcoholic subjects. Also, there was a 76% reduction in ethanol stimulation of cAMP accumulation in lymphocytes from alcoholics. Similar results were demonstrable in isolated T cells. Unlike other laboratory tests examined, these measurements appeared to distinguish alcoholics from normal subjects and from patients with nonalcoholic liver disease. Reduced basal and adenosine receptor-stimulated levels of cAMP in lymphocytes from alcoholics may reflect a change in cell membranes due either to chronic alcohol abuse or to a genetic predisposition unique to alcoholic subjects. PMID- 3029781 TI - Protracted treatment with diazepam increases the turnover of putative endogenous ligands for the benzodiazepine/beta-carboline recognition site. AB - DBI (diazepam-binding inhibitor) is a putative neuromodulatory peptide isolated from rat brain that acts on gamma-aminobutyric acid-benzodiazepine-Cl- ionophore receptor complex inducing beta-carboline-like effects. We used a cDNA probe complementary to DBI mRNA and a specific antibody for rat DBI to study in rat brain how the dynamic state of DBI can be affected after protracted (three times a day for 10 days) treatment with diazepam and chlordiazepoxide by oral gavage. Both the content of DBI and DBI mRNA increased in the cerebellum and cerebral cortex but failed to change in the hippocampus and striatum of rats receiving this protracted benzodiazepine treatment. Acute treatment with diazepam did not affect the dynamic state of brain DBI. An antibody was raised against a biologically active octadecaneuropeptide (Gln-Ala-Thr-Val-Gly-Asp-Val-Asn-Thr-Asp Arg-Pro-Gly-Leu-Leu-Asp-Leu-Lys ) derived from the tryptic digestion of DBI. The combined HPLC/RIA analysis of rat cerebellar extracts carried out with this antibody showed that multiple molecular forms of the octadecaneuropeptide-like reactivity are present and all of them are increased in rats receiving repeated daily injections of diazepam. It is inferred that tolerance to benzodiazepines is associated with an increase in the turnover rate of DBI, which may be responsible for the gamma-aminobutyric acid receptor desensitization that occurs after protracted benzodiazepine administration. PMID- 3029782 TI - Aldosterone activates Na+/H+ exchange and raises cytoplasmic pH in target cells of the amphibian kidney. AB - The hypothesis was tested if the mineralocorticoid hormone aldosterone stimulates Na+/H+ exchange in "giant cells" fused from individual target cells of the distal nephron of the frog kidney. By means of microelectrodes, steady-state intracellular pH (pHi) and pHi recovery from an acid load were recorded continuously while the fused cells were exposed to aldosterone. Twenty minutes after addition of the hormone, pHi started to rise and reached a new steady state after about 60 min (delta pHi = 0.28 +/- 0.01). After hormone treatment, pHi recovered significantly faster in response to an intracellular acid load. The diuretic drug amiloride blocked pHi recovery. Experiments in intact tubules showed that aldosterone induces H+ and K+ secretion. Thus, intracellular alkalosis, mediated by Na+/H+ exchange, could serve as a signal that activates pH sensitive K+ channels of the luminal cell membrane. PMID- 3029783 TI - The relationship between glucagon receptors and metabolic profile in two strains of rats: Wistar-Furth and Sprague-Dawley. AB - We studied glucagon and insulin binding to isolated hepatocyte receptors in Wistar-Furth (WF) and Sprague-Dawley (SD) rats, using 125I-labeled hormones. Hepatocytes from WF rats bound more glucagon than hepatocytes from SD rats. There were no differences in insulin binding. These observations prompted us to investigate other strain differences. Fasting and nonfasting serum glucose, glucagon, insulin, and growth hormone were measured. WF animals had a lower fasting glucose and higher fasting glucagon than SD animals, while SD rats had higher nonfasting insulin levels and a higher hepatic glycogen content. Total hepatic glucose production in response to glucagon (10(-8) M) was greater in WF than in SD rats, while glucagon-stimulated gluconeogenesis from alanine was the same in the two groups of animals. We concluded that the decreased glucagon binding does not play a significant role in the maintenance of serum glucose or in the gluconeogenetic response glucagon, and that neither these responses nor the serum glucagon levels appears to be correlated with the number of glucagon receptors. We conclude further that different animal strains of the same species may differ in their biologic responses. PMID- 3029784 TI - Antiherpetic action of prostaglandin D2. AB - Several prostaglandins including PGE1, PGE2 and PGF2 alpha affect the course of viral infection. There is no report about the effect of PGD2 on virus infections, although there are reports about its antitumor effect. In the present study, we examined the effect of prostaglandin D2 (PGD2) on the growth of herpes simplex virus (HSV) type 1 in human amnion FL cells. HSV yields were inhibited when FL cells were treated with PGD2 and infected with HSV at various multiplicity of infection (MOI). Treatment with PGD2 after virus adsorption inhibited HSV yields, although treatment of FL cells with PGD2 before virus adsorption did not inhibit them. PGD2 reduced the mortality of the mice infected with HSV. In conclusion, it is suggested that PGD2 has an antiherpetic activity in vitro and in vivo. PMID- 3029786 TI - [Current information on benzodiazepine-GABA-receptor complexes--perspectives on the development of highly specific neuropsychotropic drugs]. PMID- 3029785 TI - The effect of leukotrienes on porcine alveolar macrophage function. AB - The ability of the synthetic leukotrienes LTB4, LTC4, LTD4 and LTE4 to stimulate porcine AM was compared with that of two known AM stimuli: zymosan, a particulate stimulus and phorbol myristate acetate (PMA), a soluble stimulus. The criteria for AM stimulation were: increased generation of superoxide anion (O2-), the release of the lysosomal enzymes N-acetyl-beta-D-glucosaminidase (NABG) and arylsulfatase and an increase in surface free energy. Whereas zymosan and PMA both stimulated AM according to each of the three criteria, the effect of leukotrienes was relatively minor. LTB4 (10(-6) M) and LTD4 (10(-6) M) caused a modest reduction in spontaneous O2- release by AM. LTD4 (10(-6) M) also caused a small (18%), but significant (p less than 0.05), reduction in the additional O2- release induced by PMA. LTC4 and LTE4 did not alter spontaneous O2- release. Neither spontaneous nor stimulated enzyme release was systematically altered by any of the leukotrienes. LTD4 caused a cysteine-inhibitable, dose-dependent increase in surface free energy with a plateau at concentrations above 10(-8) M. The increased surface free energy induced by LTD4 may be a consequence of binding to a surface dipeptidase. PMID- 3029787 TI - Cancer presenting as mental illness. PMID- 3029788 TI - Herpes simplex virus, cytomegalovirus and Epstein-Barr virus antibody titres in sera from schizophrenic patients. AB - Serum antibody titres to herpes-simplex (HSV-1, 2), cytomegalovirus (CMV), and Epstein-Barr virus capsid antigen (EBV-VCA) were determined in 38 unrelated chronic schizophrenic patients, 11 nuclear families with at least 2 schizophrenic members, and 2 control groups. The distributions of antibody titres to herpes simplex and cytomegalovirus were similar among all groups. Patients had higher anti-EBV-VCA titres than non-hospitalized controls; however, hospital staff members in contact with the patients also had significantly higher antibody titres to EBV-VCA. Antibodies to EBV early antigen (EBV-EA) were also determined for 27 unrelated patients and 24 mental hospital employees. The schizophrenic patients had significantly higher antibody titres to EBV-EA than the hospital worker control group. These data do not support the hypothesis that herpes viruses are associated with the aetiology of schizophrenia. Although elevated anti-EBV early antigen titres may suggest persistent active EBV infection, it is unlikely to be related to the aetiology of the disorder, since discordance for EBV seropositivity was present among sibling pairs concordant for schizophrenia. PMID- 3029789 TI - Binding of yohimbine and imipramine to platelets in depressive illness. AB - Radioligand binding to intact platelets was carried out in antidepressant-free patients and the 1 mg dexamethasone suppression test (DST) was performed. There were no differences in binding characteristics between patients and controls for either [3H]yohimbine or [3H]imipramine. There were no differences in binding between patients classified as endogenous using the Newcastle Scale, compared with non-endogenous patients, and no difference between DST suppressors and non suppressors. The severity of depression did not affect binding values. After 4-6 weeks antidepressant treatment [3H]yohimbine binding was significantly reduced but [3H]imipramine binding was unaffected. PMID- 3029791 TI - Exposure to low doses of the environmental chemical dieldrin causes behavioral deficits in animals prevented from coping with stress. AB - Experiments were conducted which assessed the effects of low doses of an environmental contaminant in conjunction with various forms of stress. Rats were given acute doses (0, 0.5, 1.5, 4.5 mg/kg) of the chemical dieldrin and subsequently exposed to a series of 40 escapable shocks, identical inescapable shocks, or no shock in an operant chamber. Eight hours later, the subjects were re-exposed in a shuttlebox to footshock which was escapable upon performance of an FR-2 shuttle response. Escape deficits which were related in magnitude to the size of the dieldrin dose were found in the inescapable shock group but not in the escapable shock or no shock groups. The data suggest that experience with the lack of control over stress is critical in determining the behavioral effects of the agent and that the behavioral effects caused by uncontrollable stress may be exacerbated by concurrent exposure to such compounds. These results are discussed in terms of previous studies on the behavioral actions of dieldrin, the response to uncontrollable stress and the common neuronal systems that may be involved. PMID- 3029790 TI - Impaired growth hormone response to sodium valproate in normal aging. AB - Evidence has been provided for impaired neurotransmitter functioning in the brain of elderly subjects. In order to assess central GABAergic transmission, the activity of the hypothalamic GABA system may be investigated by basal growth hormone (GH) response to the GABAergic drug sodium valproate (SV). For this purpose 15 healthy men (aged 19-81 years) received orally 800 mg SV or placebo tablets on two different occasions, 1 week apart. Blood samples were collected before and after drug administration for determining GH and SV plasma levels. A clear-cut increase in plasma GH was observed following SV (P less than 0.001 in young persons, P less than 0.005 in old subjects), but in the aged subjects this rise was statistically lower than in the young men (P less than 0.001 at t = 90 min). No difference was observed in basal GH levels and in SV plasma concentrations between elderly and young subjects. delta GH (= maximum post-SV GH level minus baseline GH value) was significantly inversely related to age (r = 0.90, P less than 0.001). These results may suggest an impaired hypothalamic pituitary responsiveness to a pharmacological challenge enhancing endogenous GABA tone in the elderly. PMID- 3029793 TI - Discriminative stimulus properties of (+)-N-allylnormetazocine in the rat: correlations with (+)-N-allylnormetazocine and phencyclidine receptor binding. AB - Recent studies have identified a stereospecific (+)-NANM binding site that binds psychotomimetic opioids and phencyclidine (PCP) but has a distribution in brain different from the PCP binding site. Since (+)-NANM has no opioid activity and ( )-NANM has opioid activity, rats were trained to discriminate (+)-NANM from saline in order to develop an ability to distinguish the (+)-NANM cues from other opioid agonist and antagonist activities. Cyclazocine, PCP, and ketamine all produced (+)-NANM-like stimuli in a dose-dependent manner. Behaviorally, cyclazocine and PCP are equipotent to (+)-NANM whereas ketamine is 6.7 times less potent than (+)-NANM. Pentazocine had the highest affinity for the (+)-[3H]NANM binding site, yet did not produce (+)-NANM-like discriminative stimuli. By contrast, ketamine had the lowest binding affinity for the (+)-[3H]NANM binding site and did produce (+)-NANM-like discriminative stimuli. Drug discrimination potencies relative to (+)-NANM were not predictive of relative binding affinities at (+)-NANM or PCP binding sites, although there was a trend toward a stronger correlation with the PCP binding site. Therefore, the discriminative stimulus properties of (+)-NANM cannot be explained by pharmacologic actions at either (+) NANM or PCP binding sites alone, and may involve concurrent actions at both sites. PMID- 3029792 TI - Increased susceptibility to memory intrusions and the Stroop interference effect during acute marijuana intoxication. AB - The marijuana-induced acute memory impairment was assessed in a double-blind, crossover experiment. Twelve males smoked NIDA-supplied cigarettes containing 1.2% delta-9-tetrahydrocannabinol (THC) or cannabinoid-exhausted marijuana (placebo) in counterbalanced order on 2 days 1-3 weeks apart. Practice, pre- and postsmoking test sessions were conducted with the Paced Auditory Serial Addition Test, Stroop Color and Word Test, and alternate forms of the Randt Memory Battery and the Controlled Oral Word Association Test. A significantly greater number of short story omissions and intrusions occurred in delayed free recall after marijuana. Immediate and sustained attention, controlled retrieval from semantic memory, and speed of reading and naming colors were not affected. The Stroop interference effect was significantly greater following marijuana. Subjects appeared to experience parallel difficulties in inhibiting associations to the new material and inhibiting the overlearned response of reading in a new learning task. Marijuana may compromise associative control, presumably a cognitive process inherent in memory function. PMID- 3029794 TI - Conditioning of pre- and post-synaptic behavioural responses to the dopamine receptor agonist apomorphine in rats. AB - We investigated whether pharmacological effects of the dopamine agonist apomorphine can be conditioned by establishing an association of apomorphine administration with exteroceptive cues. Apomorphine was repeatedly administered and subsequently, the rat was put into a test cage and exposed to an acoustic and an olfactory stimulus ("conditioned rats"). Control animals ("pseudoconditioned" rats) were treated with the same pharmacological schedule of apomorphine not temporally associated with the stimuli. On the test day, both groups were injected with saline and exposed to the stimuli described. The stereotyped behaviour produced by large doses of apomorphine (0.5 or 2.0 mg/kg SC), namely sniffing, licking and gnawing, could be conditioned in a pronounced way. During the conditioning period, a change in the stereotypies was observed with regard to the time-course (earlier occurrence) and to the character of the stereotypies (from sniffing to licking and gnawing), when 0.5 mg/kg apomorphine was used, but not with the dose of 2.0 mg/kg. The conditioned responses showed a relatively uniform distribution during the observation period with some increase towards the end of the observation period. Some signs produced by a low dose of apomorphine (0.07 mg/kg SC), namely hypomotility and ptosis, but not yawning, could also be conditioned, although in a less pronounced way. An intermediate dose of apomorphine (0.18 mg/kg SC) produced both signs observed after large doses and those observed after a small dose, occurring alternatingly. Both types of signs could be conditioned using this dosage. Conditioning did not alter striatal or mesolimbic dopamine turnover.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3029796 TI - Marital quality, marital disruption, and immune function. AB - Marital disruption is associated with significant increases in a variety of psychologic and physical disorders. In order to examine psychologic and physiologic mediators, self-report data and blood samples were obtained from 38 married women and 38 separated/divorced women. Among married subjects, poorer marital quality was associated with greater depression and a poorer response on three qualitative measures of immune function. Women who had been separated 1 year or less had significantly poorer qualitative and quantitative immune function than their sociodemographically matched married counterparts. Among the separated/divorced cohort, shorter separation periods and greater attachment to the (ex)husband were associated with poorer immune function and greater depression. These data are consistent with epidemiologic evidence linking marital disruption with increased morbidity and mortality. PMID- 3029795 TI - Proconflict and electrocorticographic effects of drugs modulating GABAergic neurotransmission. AB - Proconflict and electrocorticographic effects of drugs acting on the benzodiazepine (BDZ)/GABA/chloride-ionophore receptor complex were studied in rats in an attempt to correlate their anxiogenic and epileptogenic activities. Evidence for proconflict activity was assessed by means of an operant conflict procedure based on the simultaneous reward and punishment of a conditioned task, while epileptogenic properties were assessed by monitoring the electrocorticogram (ECoG) of free-moving rats. Pentylenetetrazole and picrotoxin, which act through a site on the chloride channel, and the benzodiazepine (BDZ) inverse agonist FG 7142 showed epileptogenic alterations in the ECoG at doses, respectively, 8, 2 and 3 times higher than those eliciting a significant proconflict effect. For the partial inverse agonist CGS 8216, a ratio of about 60 was found while the BDZ antagonist Ro 15-1788 showed neither epileptogenic nor proconflict activity, except at the highest tested dose for the latter effect (40 mg X kg-1 PO). Inhibition of GABA transmission may mediate both anxiogenic and epileptogenic actions, and a link between these properties may exist as a continuous spectrum of negative intrinsic efficacy at the central BDZ/GABA/chloride-ionophore receptor complex. PMID- 3029798 TI - [Effect of ionizing radiation on O2 generation by cytochrome P-450]. AB - A study was made of generation of superoxide anion-radical (O2-) by cytochrome P 450 in a microsomal membrane of rat liver. Using spectrophotometry (by oxidation of adrenaline to adrenochrome) and ESR (with a spin-trap, tiron) the authors showed the ability of O2- generation by P-450 through decomposition of organic peroxides. During the first 24 h following irradiation of rats with doses of 7 and 10 Gy, the generation of O2- by cytochrome P-450 of rat liver microsomes was increased. Mechanisms of the postirradiation modification of O2- generation rate are discussed. PMID- 3029797 TI - [Thymidine excretion from thymocytes in irradiated and nonirradiated rats during endonucleolysis]. AB - The concentration of free thymidine (dT), excreted from thymocytes of irradiated and nonirradiated rats, was determined after incubation of cells in various digestive buffers. The release of dT from thymocytes depended upon the rate of DNA fragmentation in conditions of chromatin endonucleolysis. The increase in the thymidine content, in conditions of chromatin endonucleolysis in buffers containing no calcium ions, was only noted in thymocytes of exposed rats: this was the consequence of chromatin DNA damages already available in these cells. PMID- 3029799 TI - [Mechanism of the light flashes induced in human beings by ionizing particles]. AB - Exposure of a dark adapted human eye to weak proton beams of different energy for which the yield of Cerenkov radiation varies by approximately 50 times, the other characteristics being virtually the same, showed that this radiation was mainly responsible for visual sensation. PMID- 3029800 TI - [Use of gammaphos for protecting the brain from delayed radiation damage]. AB - The influence of a radioprotector, gammaphos, on the development of delayed vascular changes and necrosis in rat brain following local brain irradiation with 25 Gy was investigated. The radioprotective effect was manifested by both the morphometric parameters of vessels and the survival rate and relative number of animals with gross vascular abnormalities and brain necrosis. There was a causative relationship between the development of gross vascular abnormalities and the occurrence of brain necrosis after exposure to moderate radiation doses. PMID- 3029801 TI - [Effect of melanin on the free-radical states of gamma-irradiated proteins and lipids]. AB - It was shown that the concentration of paramagnetic centres of dihydroxyphenylalanine-melanin increased after gamma-irradiation (60Co) both at room temperature (an irreversible increase) and at 77 K (a reversible increase). The accumulation of paramagnetic centres in gamma-irradiated albumin at room temperature was found to slow down appreciably in the presence of melanin. This effect is thought to be associated with the antiradical activity of the pigment. PMID- 3029802 TI - [Modifying effect of ionizing radiation on the transmembrane transport of sodium ions in Ehrlich carcinoma tumor cells]. AB - The effect of a local X-irradiation on the transmembrane transfer of Na ions and activity of Na-K ATPase in Ehrlich ascites tumor cells has been investigated. Irradiation with doses of 0.05 and 0.15 C/kg are shown to change natrium ion transport. This effect is absent at a dose of 0.08 C/kg. A change in the rate of active transport correlates to some extent with the disturbance in Na-K ATPase occurring at the same radiation doses. PMID- 3029803 TI - Experimental and human brain neoplasms: detection with in vivo sodium MR imaging. AB - Elevations of intracellular sodium concentration have been observed in rapidly proliferating cells and malignant neoplasms. Sodium magnetic resonance (MR) imaging (with repetition times of 133 msec and echo times of 13, 26, 39, and 42 msec) was performed in ten patients and three dogs with central nervous system neoplasms. In all instances the neoplasms were associated with an increased sodium signal compared with that of normal brain. Unfortunately, the available echo times did not enable discrimination of intracellular sodium from extracellular sodium, which was present in high concentrations in adjacent vasogenic edema fluid. Further study is necessary to establish the utility of sodium MR imaging for the investigation of malignant neoplasms. PMID- 3029805 TI - Iodine-131-labeled lipiodol retention within hepatic cavernous hemangioma. PMID- 3029804 TI - Hepatic tumors: dynamic MR imaging. AB - Thirty-six patients with hepatic tumors (28 hepatocellular carcinomas, seven cavernous hemangiomas, one metastatic tumor) were examined with serial magnetic resonance (MR) imaging, after a bolus intravenous injection of 0.05 mmol/kg gadolinium-diethylenetriaminepentaacetic acid. A typical MR imaging pattern for hemangiomas (present in five of seven cases [71.4%]) was a lesion of diminished signal intensity on precontrast images, peripheral contrast enhancement during the bolus dynamic phase, and complete fill-in of high signal intensity on delayed scan images. Twenty-eight hepatocellular carcinomas showed a variety of contrast enhancement patterns during the dynamic phase. In 21 patients (75%), there was no area of high signal intensity within the tumor on the delayed phase. A peripheral halo with delayed enhancement was noticed in 12 patients (42.8%) Histologic correlation in hepatocellular carcinomas showed that the degree of contrast enhancement corresponded to tumor vascularity and that the peripheral halo corresponded to fibrous capsular structure. PMID- 3029807 TI - Interaction with mitochondria of the anthracycline cytostatics adriamycin and daunomycin. PMID- 3029806 TI - Small hepatocellular carcinoma: intratumor ethanol treatment using new needle and guidance systems. AB - Intratumor injection of absolute ethanol to treat small hepatocellular carcinoma sometimes results in incomplete necrosis of the tumor. Causes of this include inhomogeneous distribution of the ethanol and difficulty in identifying the tumor after previous ethanol injections. To solve these problems, the authors designed a multiple-side-hole needle for ethanol injection and implanted one or more small steel coils into the tumor before treatment to serve as a landmark. Six patients thus treated all showed adequate necrosis on follow-up computed tomography, biopsy, and angiography studies; initially elevated serum alpha-fetoprotein levels present in five patients were decreased. A resected surgical specimen obtained in one patient showed extensive necrosis of the tumor as well as of the surrounding healthy liver; only a small locus of equivocally viable cancer cells remained in the tumor margin. PMID- 3029808 TI - [Mechanism of electrophoration: basis of electric-pulse mediated gene transfer]. PMID- 3029809 TI - Prostaglandin effect on the physical properties of gastric mucus glycoprotein and its susceptibility to pepsin. AB - The effect of 16,16-dimethyl prostaglandin E2 (DMPGE2) on gastric mucus glycoprotein viscosity, permeability to hydrogen ion and degradation by pepsin was investigated. Preincubation with DMPGE2 produced a marked enhancement in the glycoprotein viscosity. The increase was concentration dependent and at 2.6 X 10( 5)M DMPGE2 reached a value of 178%. Permeability measurements revealed that 2.6 X 10(-7)M DMPGE2 increased the retardation ability of the glycoprotein to hydrogen ion by 10%, while 22% increase was obtained with 2.6 X 10(-4)M DMPGE2. The results of peptic activity assay showed that DMPGE2 had no inhibitory effect on the rate of glycoprotein proteolysis, and actually a small stimulatory influence was consistently observed. The results suggest that prostaglandins beneficially affect the physical properties of mucus glycoprotein which are considered to be essential for the protective function of gastric mucus. PMID- 3029811 TI - GABA and cardiovascular regulation. PMID- 3029810 TI - Comparison of the urinary metabolites of prostacyclin and thromboxane of the 2- and 3-series in a Japanese fishing and a Japanese farming village. AB - We measured PGI2-, PGI3-, and TXA2/3-M (the main urinary metabolites of prostacyclin and thromboxane of the two and three series) in 24-hour urine in a fishing village (21 participants) and a farming village (19 participants) in Japan, expecting to find more PGI3-M in the fishing village than in the farming village. The food consumption for three consecutive days prior to a blood collection was recorded and analyzed for eicosapentaenoic acid (EPA) content. Urine was collected for 24 hours prior to the blood sampling. When our studies were performed (April, 1985), catches of fish were the lowest on record around the fishing village, and the consumption of EPA in the fishing village was less than half of that in the farming village. EPA levels in red cell membrane phospholipids were higher in the fishing village than in the farming village. PGI2-, PGI3-, and TXA2/3-M levels were higher in the farming village than in the fishing village. There was a significant correlation between PGI2- and PGI3-M (r=0.80, n=40) and between PGI3-M and the EPA consumption during the day of urine collection (r=0.52, n=40). We conclude that PGI3 production is probably dependent less on the levels of EPA in tissues estimated by the EPA levels in red cell membranes than on the EPA consumption at the moment. PMID- 3029812 TI - Aging: effect on the biochemistry of beta-adrenergic catecholamine action. PMID- 3029813 TI - Regulation of contractile activity in vascular smooth muscle by protein kinases. AB - Control of the contraction/relaxation cycle in vascular smooth muscle is regulated by Ca2+ and the cyclic nucleotides, cAMP and cGMP. For the most part, the effectors of these intracellular messengers are the protein kinases. Four major protein kinases (myosin light chain kinase, protein kinase C, cAMP dependent protein kinase, and cGMP dependent protein kinase) have been identified in vascular smooth muscle. Substantial biochemical and physiological evidence exists supporting the involvement of Ca2+/calmodulin-mediated activation of myosin light chain kinase and phosphorylation of the 20,000 dalton P-light chain of myosin in the regulation of vascular contractile activity. However, alternative hypotheses exist which suggest that additional Ca2+ dependent regulatory mechanisms reside at other contractile protein sites. Calcium also activates protein kinase C, which requires phospholipid and diacylglycerol as co factors instead of calmodulin. Protein kinase C also phosphorylates smooth muscle myosin P-light chain; however, phosphorylation occurs at a different site on the P-light chain and represses ATPase activity which has been stimulated by myosin light chain kinase-catalyzed phosphorylation. The precise physiological role of protein kinase C in modulating vascular smooth muscle contractile activity remains to be elucidated. Relaxation of vascular smooth muscle by some different relaxants is linked to either cAMP or cGMP formation. Correlative evidence also links activation of cAMP dependent protein kinase with relaxation. Two isozymes of cAMP dependent protein kinase exist in arterial smooth muscle; potential specific roles for each isozyme have not been elucidated. Mechanistically, relaxation mediated by both cyclic nucleotide-regulated protein kinases most likely involves primary effects on Ca2+ ion flux regulation rather than direct effects on contractile protein interactions. Activation of cGMP dependent protein kinase may be important in mediating the relaxant effects of endothelium derived relaxant factor or atrial natriuretic factor. Direct pharmacological modulation of smooth muscle vascular protein kinase activity represents an approach towards developing novel vasodilator agents. Various classes of agents, including phenothiazine antipsychotics, antidepressants, naphthalene sulfonamides, and certain lipophilic Ca2+ antagonists, inhibit myosin light chain kinase activity primarily by competition with the enzyme for Ca2+-calmodulin. However, additional inhibition via binding to the myosin P-light chain may also occur with some of these agents.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3029814 TI - Phospholipid turnover and gonadotropin releasing hormone action in the pituitary. PMID- 3029815 TI - Phosphoinositides and signal transductions. PMID- 3029816 TI - Possible involvement of phosphoinositide phosphorylation and tyrosine kinase activity in the growth control of rat mammary tumors. PMID- 3029818 TI - [Physiopathology of the panic syndrome]. PMID- 3029819 TI - [Calciuria, calcium overload and urinary cyclic AMP in the evaluation of patients with calcic lithiasis]. PMID- 3029817 TI - The biochemical functions of pp60v-src. PMID- 3029820 TI - [Mixed tumor of the lacrimal gland]. PMID- 3029822 TI - [Subungual glomus tumor. A case of unusual form]. AB - The authors report a case of glomus tumour developing beneath the nail of the ring finger in a 95 year old woman. In contrast with other similar cases, the pain was moderate and had been present for 20 years. The bony phalanx was deformed. Microscopic examination confirmed the diagnosis of a glomus tumour developing in the soft tissues. The phalanx was amputated. PMID- 3029821 TI - [Tumors of the glomus jugulare. Comments on 3 personal cases]. PMID- 3029823 TI - [Imaging of neoplastic changes in the spine using MR tomography]. AB - The value of MR tomography for examining vertebral infiltration by malignant disease and the use of various techniques was examined retrospectively in a material of 68 examinations in 62 patients suffering from metastases, plasmacytomas, primary vertebral tumours and vertebral infiltration from extra osseous tumours. It was shown that the method is superior to all other imaging methods for demonstrating changes in the bone marrow. The extent of the disease and involvement of neighbouring structures is shown more completely in cases where the disease extends beyond the vertebrae. Improved soft tissue contrast and the availability of various planes are definite advantages, as compared with CT. Tissue diagnosis, however, is not improved by MRT. PMID- 3029824 TI - [Value of magnetic resonance tomography prior to surgery and irradiation of peripheral soft tissue tumors]. AB - The experience with magnetic resonance imaging (MRI) of 37 patients with peripheral soft tissue tumours and tumour-like lesions is reported. MRI proved to be a highly sensitive method in detecting soft tissue tumours. The main advantages of MRI compared to CT were the high contrast resolution of MRI and the multiplanar display. The anatomic setting of the soft parts tumour could be defined in most cases as either intracompartmental or extracompartmental. The relationship to the major neurovascular bundles and bone could be assessed accurately. Tumour-induced subtle periosteal reactions and cortical erosions are better defined with CT and plain film, especially. An imaging technique using small excitation angles, echoes produced by gradient inversion and extremely fast repetition times ("Fast Field Echo Imaging") is capable of producing MR images in only a few seconds acquisition time. Preliminary experience shows that soft tissue tumours can be evaluated with this rapid pulse sequence. PMID- 3029825 TI - [31P NMR spectroscopy of muscular diseases: correlation with MR imaging]. AB - The spectra of normal and abnormal muscle were examined by 31P MR spectroscopy using a 2 Tesla system. Exercise tests on normal and abnormal muscle were also carried out. Abnormal muscles showed characteristic changes in the phosphorus spectrum, when compared with normal muscles, both at rest and after exercise; these were compared with the data from the MR images. The advantages and limitations of the method are discussed. PMID- 3029826 TI - [31-phosphorus spectroscopy of osteogenic sarcoma. Possibilities in therapy control in comparison with imaging methods]. AB - 31-phosphorus spectroscopy was used to control the results of chemotherapy in a patient with an osteogenic sarcoma. The findings were compared with conventional imaging methods (DSA, MRT and conventional radiography). The imaging methods do not provide reliable information regarding growth of tumour tissue during treatment, whereas the determination of phosphate concentration in the tumour provides direct information concerning tumour metabolism. PMID- 3029827 TI - [Improvement in the informativeness of diuretic kidney function scintigraphy by calculation of a washout index]. AB - Diuresis renography after administration of 123I-(131I-)Hippuran of 248 patients (492 kidneys) were compared retrospectively with clinical findings, history, i.v. urography, and ultrasound examinations. A new wash-out-index was calculated. In cases with obstructive dilatation of the collecting system this index was smaller than 0.9. If it was 0.9 to 1.2 additional diagnostic procedures seem necessary. An index more then 1.4 excludes obstruction. The shape of the renogram curve after furosemide is important for the diagnosis too. A concave shape was found in non-obstructive dilatation, a convex shape in obstructive dilatation of the renal pelvis. The wash-out-index is a reliable parameter if cases with reflux, lower ureteral obstruction and shrunken kidneys are excluded. PMID- 3029828 TI - [Nuclear magnetic resonance tomographic studies following experimental ligation of the renal artery in rats]. AB - MR was carried out on 22 Han-Wistar rats in relation to ligature of the left renal artery; measurements were obtained before the operation and at two, four, eight and thirty-six hours afterwards. Significant increase in T2 relaxation of the left cortex was observed after thirty-six hours and increased T2 of the medulla on the operated side after two hours. T1 relaxation time in the cortex was also prolonged, but the medulla showed no definite change. Following the operation, there was an increase in the size of the opposite kidney after eight hours; on the operated side, the cortex increased, but the medulla decreased. The results indicate that MR is a very sensitive method for demonstrating renal artery occlusion at an early stage. PMID- 3029830 TI - [False-positive findings of 67-gallium scintigraphy in staging/restaging of malignant lymphoma]. AB - Gallium scintigraphy has been an established method for the diagnosis of malignant lymphomas for some years. If properly interpreted, the value of the method is not necessarily reduced by the lack of tumour specificity and the well known property of 67Ga to accumulate in inflammatory and granulomatous conditions. A retrospective analysis of 123 patients with malignant lymphomas showed false positive 67Ga scintigrams in 7%. These cases demonstrate the spectrum of possible errors. 'False positives' were all those localised areas of uptake which were due to some pathological condition which was not lymphomatous. All the positive gallium findings could be elucidated by clinical, radiological and histological investigations. PMID- 3029829 TI - [Results and complications of percutaneous nephropyelostomy in 215 patients]. AB - The results of 242 percutaneous nephropyelostomies in 215 patients were analysed retrospectively. No consideration was given to the experience of the individual operators. There was a total complication rate of 36.4% (mild complications 32.2%, serious complications 34.1%, fatality 0.4%). Rapid reduction in serum creatinine could be demonstrated following drainage in patients in whom renal function had been damaged by obstruction. PMID- 3029831 TI - [Focal changes in the spleen. Ultrasonic morphological characteristics and their clinical significance]. AB - Focal changes in the spleen were rare findings in a large clinical material (less than 1% of cases). In a prospective study, which included 580 patients, lesions in the spleen were found in 40. Four focal lesions were due to infiltrates from non-Hodgkin's lymphoma. Twenty-one lesions with low echoes consisted of twelve infiltrates from Hodgkin's disease (six patients) or non-Hodgkin's lymphoma (six patients), five were due to fresh splenic infarcts and one each to an abscess, a metastasis from a carcinoma of the stomach, sarcoid and a haemorrhage. In only three of ten highly echogenic foci was a diagnosis possible (one leukaemic infiltrate and two scars following splenic infarcts). The significance of the sonographic demonstration of focal splenic lesions for diagnosis and treatment is discussed. The advantages of a sector scanner for evaluation of the spleen and splenic size are mentioned. PMID- 3029832 TI - [The variable picture of gastric carcinoid]. AB - Carcinoids are endocrine tumors which develop from enterochromaffin cells and will be found in more than 80% in the gastrointestinal tract. Only 2-3% of these carcinoids is located in the stomach. The rarity of their occurrence and the wide variety of radiographic features (intramural defects, multiple gastric polyps, large gastric ulcers and polypoid intraluminal lesions) make their recognition difficult. We report 3 cases of gastric carcinoid, one of them presented with pernicious anemia, which has been reported more frequently in literature. One patient showed the unusual combination of adenocarcinoma and a carcinoid with a stalk in the stomach. PMID- 3029834 TI - [Imaging of femur head necrosis using MR tomography]. AB - Magnetic resonance tomography (MR) was used to examine the femoral heads of 20 normals and of 56 patients with diseases of the femoral head, including Perthe's disease (four cases), adult necrosis of the femoral head (39 cases) and pain of unknown cause (13 cases). Femoral head necrosis and Perthe's disease regularly produce reduced signal intensity. The localisation of the areas of necrosis can be determined accurately in the coronary and sagittal plane. In 11 femoral heads, necrosis could be demonstrated unequivocally by MR in the presence of normal radiographic findings. Follow-up of nine patients with negative MR findings and indefinite symptoms confirmed the absence of necrosis. PMID- 3029833 TI - [Is hip sonography meaningful as a screening method?]. AB - The results of screening 162 non-selected neonates by sonography of the hips indicate that this is not a desirable screening method. Sonography is, however, indicted in neonatal groups which are considered to be at risk. PMID- 3029836 TI - [Soft tissue changes in the hands in progressive scleroderma (excluding calcifications)]. AB - The systemic progressive sclerosis causes numerous soft tissue changes at the hand, most often a thickening of the skin and an augmentation of the fibrous structures in the subcutis, which occur before the atrophy of the finger tip. Other soft tissue changes are the synovitic enlargement of the joint capsules and the tendon sheaths and the tendinitic enlargement of the tendons. This complex of soft tissue changes is seen in the three view low kV radiography. PMID- 3029835 TI - [Angiographic control of treatment of femur head necrosis with a pedicled pelvic bone graft]. AB - A new operative method is introduced in the therapy of the idiopathic necrosis of the femoral head. This operation is based on the idea, that ischaemic processes play an important role in the origin of the idiopathic necrosis of the femoral head. The transplantation of a pedicle bone graft should provide a good arterial supply. 28 Preoperative superselective angiographies of the A. circumflexa femoris medialis showed pathological findings in 17 cases. A good supply of the bone graft was found in 4 from a total of 7 postoperative angiographies of the reset A. circumflexa ilium profunda. PMID- 3029837 TI - [Characteristics of intracranial meningioma imaged by magnetic resonance tomography]. AB - Twenty-three patients with intra cranial meningiomas were examined by means of magnetic resonance tomography (MRT). In 13 patients the paramagnetic contrast medium gadolinium DTPA was used. Meningiomas show only slight changes in signal intensity compared with brain in the spin-echo mode, the greatest contrast being found on photon density images (TR 1600 ms, TE 35 ms). In T1 images, more than 50% patients showed a low signal margin between tumour and brain, Hyperostosis of the calvarium is easily recognised, but MRT is unreliable for showing tumour calcification. After intravenous injection of gadolinium DTPA, there was marked homogeneous uptake in the meningiomas. These signs are useful for the diagnosis of a meningioma by MRT. PMID- 3029838 TI - [Rapid nuclear magnetic resonance tomography. Initial results of studies using the new gradient echo sequence]. AB - In 60 patients with intracerebral lesions, examined by MRI, a new gradient-echo sequence was employed. This imaging technique uses excitation pulse angles smaller than 90 degrees and echoes are produced by an inversion of the read gradient. Since no 180 degrees pulse between successive excitations is necessary, very short repetition times can be used. Depending on matrix-size and signal averaging, MR imaging time can be reduced to approximately 5 to 40 seconds for single slice scan. For comparison, conventional T1- and T2-weighted spin-echo images were performed. Diagnostic results of the T1-weighted fast gradient-echo images corresponded with T1-weighted spin-echo images in 30% of cases. Diagnostic information was lower in 40% of cases. In the remainder of cases (30%) lesions were not detected with the gradient-echo technique. This especially applied to multiple sclerosis, infarctions and low-grade gliomas. Due to image artefacts and low contrast, visualization of small pathologic lesions was limited. Significant improvement of tumor visualization on gradient-echo scans was observed after injection of Gd-DTPA (0.1 mmol/kg). PMID- 3029839 TI - [A simple attachment for superficial sonography]. AB - Electronic linear array transducers usually have a discrepant electronic (in scanning plain) and mechanical (perpendicular to scanning plain) focal zone. The "effective" focal zone can only be determined with a true cyst phantom. With transducers of medium frequency the focal zone is about 3-6 cm from the transducer surface, rendering a water path necessary for sonography of superficial organs. The considerable thickness of layer in the scanning plain of most linear array transducers affects the visualisation of superficial vessels in longitudinal direction. A simple water path attachment for small parts sonography is presented, which enables an automatic "self-indication" of palpable lesions especially in breast sonography. PMID- 3029840 TI - [Diagnosis of duodenal wall cysts. A case report]. PMID- 3029841 TI - [Choledochal diverticulum as a possible cause of right-sided upper abdominal pain]. PMID- 3029842 TI - [Myelolipoma of the adrenal glands. Correlation of radiologic and pathologico anatomic characteristics]. PMID- 3029843 TI - [Congenital peripheral arteriovenous fistulas]. PMID- 3029845 TI - Intranasal vaccination of pigs against Aujeszky's disease: comparison with one or two doses of attenuated vaccines in pigs with high maternal antibody titres. AB - Ten-week-old pigs with high levels of maternally derived antibody (MDA) against Aujeszky's disease virus (ADV) were given either a single intranasal vaccination or one or two doses (with an interval of three weeks) of commercially available attenuated ADV vaccines intramuscularly. The pigs did not produce a clear neutralising antibody response to ADV. However, pigs vaccinated intranasally and pigs given two doses of attenuated ADV vaccines were protected against intranasal challenge with virulent ADV two months after the first vaccination. Pigs given one parenteral dose of attenuated ADV vaccine were insufficiently protected. Protection was shown by shorter periods of growth arrest and fever and a greater reduction of virulent virus shedding after challenge in vaccinated pigs than in unvaccinated control pigs. Although intranasal vaccination conferred protection comparable to two parenteral doses of attenuated vaccines, it reduced shedding of virulent virus much more effectively. These results, together with those of other studies, show that intranasal vaccination confers better protection against Aujeszky's disease in pigs with MDA than parenteral vaccination. However, the efficacy of intranasal vaccination also decreases with increasing levels of MDA at the time of vaccination. PMID- 3029844 TI - [Bronchiolitis and shock: biochemical monitoring]. PMID- 3029846 TI - Virulence of bluetongue virus for British sheep. AB - A South African isolate of bluetongue virus type 3 was inoculated intradermally into three different breeds of British sheep under conditions designed to test its virulence in animals under stress. All animals inoculated developed a pyrexia and viraemia followed by clinical evidence of bluetongue disease. Marked alterations in serum enzyme levels, in particular of creatine phosphokinase, lactate dehydrogenase and aldolase occurred in the more severely affected animals. Nine out of the 12 inoculated animals subsequently died. No major differences in response could be detected in the different breeds of sheep nor in the stressed compared with the unstressed groups. The virulence of this bluetongue virus isolate was thereby confirmed and its potential risk to the British sheep industry. Consequently, stringent import regulations must be maintained to prevent its entry into Britain. PMID- 3029847 TI - Measurement of volatile fatty acid production rates in sheep given roughage. AB - Measurement of the irreversible loss rate (ILR) of acetate, propionate and butyrate by radioisotope dilution of the respective tracers in sheep offered chopped dried grass from continuous belt type feeders gave 'plateau samples', the specific radioactivity of which had unacceptably high variance (coefficient of variation [cv] of the five or six samples taken from any one animal over the last four hours of 10 hours infusion ranged from 16 to 57 per cent). In an experiment which attempted to identify the source of this variability the distribution of the liquid phase marker 51Cr EDTA into different areas of the rumen was examined during its administration either in liquid form or impregnated on paper. The paper provided better mixing but incurred considerable spillage problems which would be unacceptable where calculations of ILR were required. Radiographic observations were made to examine the mixing of radio-opaque liquid and solid phase markers in the rumen. A better mixing of both was achieved with intermittent than continuous feeding. When 1-14C- and 2-14C-labelled acetate were each infused into the rumen of four sheep given two levels of chopped dried grass by means of hourly feeders, samples of rumen liquor gave much lower variability of the mean plateau specific radioactivity values than was obtained in experiment 1 (cv less than 15 per cent in all but one sheep). The ILRs of acetate and the transfer quotients of carbon from acetate to butyrate obtained using 1-14C- and 2 14C-labelled acetate were similar.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3029848 TI - A single vaccine for the control of capripox infection in sheep and goats. AB - A single vaccine against capripox was developed to protect sheep and goats throughout the capripox enzootic area. The vaccine was stable and safe to use, and provided substantial protection against capripox for at least a year. Preliminary field trials in the Middle East indicated that the vaccine was effective in controlling capripox in indigenous breeds of sheep and goats. PMID- 3029850 TI - [Cancer of the uterine cervix: a sexually transmitted viral disease]. PMID- 3029851 TI - [Adrenergic receptors of the broncho-alveolar system]. PMID- 3029849 TI - Endurance training does not affect diaphragm mitochondrial respiration. AB - We sought to determine if chronic endurance training would increase mitochondrial respiration or protein content in rat diaphragm muscle. To this end, 20 male Wistar rats were randomly assigned to control (C) or an 8-week endurance training (T) group, n = 10 per group. At the end of T, VO2 max was 13% greater in T (83.3 vs 73.8 ml X kg-1 X min-1) and peak max power output was 32% greater (2.63 vs 1.98 kg X m X min-1). Mitochondrial specific activities of pyruvate-malate and cytochrome oxidase (expressed per mg mitochondrial protein) in both plantaris and diaphragm were similar in C and T rats, as were ADP/O and respiratory control ratios. When expressed per gram wet weight, whole muscle homogenate oxygen uptake (pyruvate + malate) and cytochrome oxidase activity increased 36 and 23%, respectively (P less than 0.05) in plantaris from T rats but did not change in diaphragm. Control oxidative capacity and mitochondrial protein content in the diaphragm were ca. 2-fold those in control plantaris. Plantaris mitochondrial protein content increased ca. 50% with T while the diaphragm was unaffected. We conclude that: plantaris muscle oxidative capacity adapts to training by increasing mitochondrial protein content, since there was no evidence for functional improvement of existing mitochondria, and in the face of a substantial training effect in whole animal and plantaris, the T stimulus was not sufficient to induce mitochondrial protein changes in the diaphragm. This finding is the result of either a 'pre-adaptation' secondary to the diaphragm's high chronic activity, or a sub-threshold increase in diaphragm recruitment during the exercise conditions studied. PMID- 3029852 TI - [Chloroquine neuromyopathies: 4 cases during antimalarial prevention]. AB - Four cases of chloroquine neuropathy presented two points of particular interest: 1) these were all young women with strictly prophylactic doses of chloroquine (100 mg/24 h); two patients showed pronounced loss of weight; 2) signs of myogenic lesions were apparent in only one patient whereas clinical and electric signs of neurogenic involvement were observed in all 4 cases. Biopsy in 3 patients (muscle in one and muscle and nerve in two) failed to show characteristic vacuoles; in one case mild segmental demyelinization was present. A favorable course was obtained in all cases after discontinuation of chloroquine, with dramatic recovery from the motor deficit. Pharmacokinetic studies in 1 patient showed abnormally high chloroquine levels at initial assay with slow elimination after its discontinuation. PMID- 3029853 TI - [Papillomavirus and cervix atypia. Retrospective histo-colposcopic study of 175 cases]. AB - One hundred and seventy five CIN cases diagnosed during the years 1958-60 have been reviewed and histologic slides as well as colpographies have been reevaluated for the presence of Papillomavirus-related features. Histo colposcopic features of HPV infection have been found in 70% of CIN cases, incidence comparable to the one actually observed. Thus, the relationship between HPVs and cervical cancer doesn't seem to be changed. PMID- 3029855 TI - Lung cancer: by the time it's detected, it may be too late. PMID- 3029854 TI - [Rotavirus reinfections in children of Belem, Para, Brazil]. PMID- 3029856 TI - [Algodystrophy in children and young adults with low isotope retention in the bones. Apropos of 5 cases]. AB - The authors report 5 observations of young adults, 3 teen-agers and 2 children suffering from algodystrophy, and in whom isotopic exploration of the skeleton disclosed a clear bony hypofixation during the entire evolution. These observations confirm their 1981 work concerning a young adult suffering from algodystrophy with isotopic bony hypofixation. Recent Canadian and American studies emphasize also the frequency of isotopic hypofixation in children algodystrophy. It seems, therefore, that isotopic bony hypofixation (linked perhaps to a decreased blood flow), is rather specific of algodystrophy in young subjects. PMID- 3029857 TI - Clinical perspectives of drugs inhibiting acid secretion--H+K+-ATPase inhibitors. AB - The H+K+-ATPase is supposed to be the terminal step in the acid-secreting pathway in the parietal cell. Omeprazole blocks this enzyme, resulting in a marked inhibition of basal and stimulated acid secretion. With omeprazole 20 mg daily, 24-hour intragastric acidity is decreased by about 90%. Several clinical studies have now been published in which omeprazole has been compared with the H2 receptor antagonists cimetidine and ranitidine. Omeprazole in doses between 20 and 40 mg daily resulted in healing rates between 65% and 82% after treatment for 2 weeks and between 90% and 100% after treatment for 4 weeks. Treatment with omeprazole also gave faster and more pronounced pain relief. One comparative study in gastric ulcer has also been published showing healing rates equal to those with ranitidine. Placebo-controlled trials have also shown very pronounced therapeutic effect in reflux esophagitis. Omeprazole seems to be the drug of choice in Zollinger-Ellison syndrome, giving beneficial clinical effects and pronounced and long-lasting reduction in gastric acid secretion. PMID- 3029858 TI - Antibiotic use as an inducer of resistant urinary tract infections. AB - Urinary tract infection is second in frequency only to respiratory infection as an indication for antimicrobial chemotherapy. As a result, a significant proportion of both domiciliary and hospital patient populations are exposed to chemotherapy which, in turn, may act as a potent selection pressure for development of resistance mediated by intrinsic mechanisms or acquisition of R plasmids or transposons. The widespread use of broad spectrum agents has encouraged the development of such resistance in enterobacteria, notably but not exclusively amongst the Klebsiella-Enterobacter-Serratia group, leading to the creation of patient reservoirs of multiply-antibiotic-resistant bacteria. Such situations occur frequently in urological units, amongst catheterised individuals receiving chemotherapy, and, via cross-infection, may rapidly spread to other units and other hospitals. The use of antibiotic combinations, such as co trimoxazole, may not prevent such problems, which are however amenable to policies which restrict, either selectively or globally, antibiotic use. Such policies can be expected to control the spread of multiply-antibiotic-resistant infection. PMID- 3029859 TI - Resistance factors in anaerobic bacteria. AB - Resistance transfer factors have been described in both Bacteroides and clostridia. The clindamycin (Cln) resistance transfer factors from the Bacteroides fragilis group of organisms have been best studied, including our own plasmid pBFTM10. The clindamycin resistance determinant (Cln X) of pBFTM10 can be detected in 90% of Cln resistant Bacteroides isolated from dispersed geographical areas. This determinant can be located in the chromosome and on plasmids. Recent studies from our laboratory have shown that the Cln X genes of pBFTM 10 are carried on a compound transposon, Tn4400. Bacteroides plasmids have been cloned in Escherichia coli and shuttle vectors have been developed that allow transfers of DNA from E. coli back to B. fragilis, using the broad host range plasmid RK2 to supply essential conjugation functions. We have shown that shuttle vectors containing pBFTM 10 can be retransferred from B. fragilis back to E. coli. In addition, a tetracycline transfer element from B. fragilis strain TM230 is able to promote high frequency conjugation between B. fragilis and E. coli. The results of these investigations indicate that Bacteroides has efficient mechanisms to exchange genetic material and that genetic exchange can occur between Bacteroides and E. coli, which exist in intimate contact in the human colon. PMID- 3029860 TI - [Vasculitic polyneuropathy as a complication in chronic polyarthritis]. AB - Polyneuropathy due to vasculitis in connection with rheumatoid arthritis is a rare complication of the disease. Special investigations can help us to distinguish between a peripheral sensory-neuropathy with a good prognosis, the neuropathy of the autonomous nervous system and the severe form of mononeuritis multiplex. The bad prognosis of this form is due to the diffuse vasculitis of internal organs. We describe 5 patients with rheumatoid arthritis and different forms of vasculitis. The differential diagnosis, therapy and prognosis are described in detail. PMID- 3029862 TI - The structure of poliovirus. PMID- 3029861 TI - [Anterior pituitary hypersecretion syndromes]. AB - Anterior pituitary hypersecretion can be due to abnormal hypothalamic regulation, decreased peripheral hormone feedback or pituitary tumor. In some cases hypersecretion gives rise to a typical clinical syndrome involving acromegaly, hyperprolactinemia, and excess corticotropin (ACTH). The etiology of acromegaly is a growth hormone (GH)-secreting pituitary tumor in the vast majority of cases. Hyperprolactinemia and excess cortisol, however, may be due to many causes among which prolactin (PRL)- and ACTH-secreting pituitary tumors are not frequent. Glycoprotein-secreting pituitary tumors, especially gonadotropin (LH and FSH) and free subunits usually do not cause a typical excess hormone syndrome. Perhaps for this reason they are seldom recognized clinically, although histopathological studies are increasingly disclosing the gonadotrope nature of many pituitary tumors. Mixed hormonal secretions are common. When pituitary hormone secretion can be selectively suppressed by medical therapy, a significant reduction of tumor size is by no means rare. In other cases, pituitary irradiation or surgery, or even treatment aimed at a peripheral target gland, may be necessary. PMID- 3029863 TI - [Sonographic diagnosis of the fluid-filled stomach]. AB - By filling the stomach with liquid, it becomes possible to represent all the sections of the wall of the stomach by means of transcutaneous ultrasonography. 68 patients were investigated in an initially orientating study. In 45 out of 53 patients with gastric wall changes, the lesion was correctly detected or correctly described. In addition to extensive wall-infiltrating processes and stenoses, circumscribed space-consuming lesions were also detected, for example, circumscribed carcinoma, leiomyosarcoma, lymphoma, leiomyoma, polyps, giant folds, gastric wall impressions, as well as early cancer and ulcers with elevated margin. Fifteen other patients with no gastric pathology were classified as unremarkable on the basis of the ultrasonographic image. The procedure would appear useful as a supplement to diagnostic endoscopy and x-rays, but also represents an alternative diagnostic procedure in selected patients who cannot be submitted to endoscopy or x-ray examination. Exclusion of any diagnosis is not possible by this method. PMID- 3029865 TI - Trans-activator gene of HTLV-II: interpretation. PMID- 3029866 TI - Molecular cloning of complementary DNA encoding the avian receptor for vitamin D. AB - Vitamin D3 receptors are intracellular proteins that mediate the nuclear action of the active metabolite 1,25-dihydroxyvitamin D3 [1,25(OH)2D3]. Two receptor specific monoclonal antibodies were used to recover the complementary DNA (cDNA) of this regulatory protein from a chicken intestinal lambda gt11 cDNA expression library. The amino acid sequences that were deduced from this cDNA revealed a highly conserved cysteine-rich region that displayed homology with a domain characteristic of other steroid receptors and with the gag-erbA oncogene product of avian erythroblastosis virus. RNA selected via hybridization with this DNA sequence directed the cell-free synthesis of immunoprecipitable vitamin D3 receptor. Northern blot analysis of polyadenylated RNA with these cDNA probes revealed two vitamin D receptor messenger RNAs (mRNAs) of 2.6 and 3.2 kilobases in receptor-containing chicken tissues and a major cross-hybridizing receptor mRNA species of 4.2 kilobases in mouse 3T6 fibroblasts. The 4.2-kilobase species was substantially increased by prior exposure of 3T6 cells to 1,25(OH)2D3. This cDNA represents perhaps the rarest mRNA cloned to date in eukaryotes, as well as the first receptor sequence described for an authentic vitamin. PMID- 3029864 TI - Bilirubin is an antioxidant of possible physiological importance. AB - Bilirubin, the end product of heme catabolism in mammals, is generally regarded as a potentially cytotoxic, lipid-soluble waste product that needs to be excreted. However, it is here that bilirubin, at micromolar concentrations in vitro, efficiently scavenges peroxyl radicals generated chemically in either homogeneous solution or multilamellar liposomes. The antioxidant activity of bilirubin increases as the experimental concentration of oxygen is decreased from 20% (that of normal air) to 2% (physiologically relevant concentration). Furthermore, under 2% oxygen, in liposomes, bilirubin suppresses the oxidation more than alpha-tocopherol, which is regarded as the best antioxidant of lipid peroxidation. The data support the idea of a "beneficial" role for bilirubin as a physiological, chain-breaking antioxidant. PMID- 3029867 TI - The regulation of natural anticoagulant pathways. AB - Vascular endothelium plays an active role in preventing blood clot formation in vivo. One mechanism by which prevention is achieved involves a cell surface thrombin-binding protein, thrombomodulin, which converts thrombin into a protein C activator. Activated protein C then functions as an anticoagulant by inactivating two regulatory proteins of the coagulation system, factors Va and VIIIa. The physiological relevance of the protein C anticoagulant pathway is demonstrated by the identification of homozygous protein C--deficient infants with severe thrombotic complications. Recent studies suggest that this pathway provides a link between inflammation and coagulation. PMID- 3029868 TI - Megabase-scale mapping of the HLA gene complex by pulsed field gel electrophoresis. AB - In the study of the genetic structure of mammalian chromosomes, there exists a "resolution gap" between molecular cloning experiments and meiotic linkage analyses. This gap has discouraged attempts to construct full-scale genetic maps of mammalian chromosomes. The organization of the human major histocompatibility complex was examined within this range by pulsed field gel electrophoresis. The data obtained indicate that the complex spans over 3000 kilobases and enable the construction of a megabase-scale molecular map. These results indicate that the techniques employed in DNA extraction, enzymatic digestion, electrophoresis, and hybridization are suitable for the efficient analysis of megabase regions of mammalian chromosomes and effectively bridge the resolution gap between molecular cloning and classical genetics. PMID- 3029869 TI - Transposition of gram-positive transposon Tn916 in Acholeplasma laidlawii and Mycoplasma pulmonis. AB - Mycoplasma genetics has been limited by a lack of genetic tools such as selectable markers, methods to transfer DNA, and suitable vectors for cloning. Studies were undertaken to examine the potential of using the streptococcal transposon Tn916 as a mycoplasma genetic tool. The Escherichia coli plasmid pAM120, which contains Tn916, was transformed into Acholeplasma laidlawii and Mycoplasma pulmonis. Transposition of Tn916 into the mycoplasma chromosome apparently occurred by an excision-insertion mechanism. This example shows that newly introduced DNA from other bacteria can be successfully expressed in mycoplasma and that Tn916 should serve as a powerful genetic tool for the study of mycoplasmas. PMID- 3029870 TI - Transformation of rat fibroblasts by FSV rapidly increases glucose transporter gene transcription. AB - Elevation of glucose transport is an alteration common to most virally induced tumors. Rat fibroblasts transformed with wild-type or a temperature-sensitive Fujinami sarcoma virus (FSV) were studied in order to determine the mechanisms underlying the increased transport. Five- to tenfold increases in total cellular glucose transporter protein in response to transformation were accompanied by similar increases in transporter messenger RNA levels. This, in turn, was preceded by an absolute increase in the rate of glucose transporter gene transcription within 30 minutes after shift of the temperature-sensitive FSV transformed cells to the permissive temperature. The transporter messenger RNA levels in transformed fibroblasts were higher than those found in proliferating cells maintained at the nonpermissive temperature. The activation of transporter gene transcription by transformation represents one of the earliest known effects of oncogenesis on the expression of a gene encoding a protein of well-defined function. PMID- 3029871 TI - The immunoglobulin octanucleotide: independent activity and selective interaction with enhancers. AB - The thymidine kinase (tk) promoter of herpes simplex virus includes an octanucleotide sequence motif (ATTTGCAT) that is also an essential component of immunoglobulin kappa gene promoters. In the absence of an enhancer, tk promoter derivatives that contain this element support a higher rate of transcription than those that lack it. The action of the kappa enhancer augments that of the octanucleotide in B lymphoid cells; when both elements are present, tk promoter activity is increased by more than an order of magnitude. In contrast, the presence of the octanucleotide in this promoter markedly reduces its response to a nonimmunoglobulin enhancer. These results suggest that the octanucleotide may mediate a selective interaction among promoters and enhancers. PMID- 3029873 TI - Oncogenesis of the lens in transgenic mice. AB - Neoplastic tumors of the ocular lens of vertebrates do not naturally occur. Transgenic mice carrying a hybrid gene comprising the murine alpha A-crystallin promoter (-366 to +46) fused to the coding sequence of the SV40 T antigens developed lens tumors, which obliterated the eye cavity and even invaded neighboring tissue, thus establishing that the lens is not refractive to oncogenesis. Large-T antigen was detected early in lens development; it elicited morphological changes and specifically interfered with differentiation of lens fiber cells. Both alpha- and beta-crystallins persisted in many of the lens tumor cells, while gamma-crystallin was selectively reduced. Accessibility, characteristic morphology, and defined protein markers make this transparent epithelial eye tissue a potentially useful system for testing tumorigenicity of oncogenes and for studying malignant transformation from its inception until death of the animal. PMID- 3029872 TI - Variable number of tandem repeat (VNTR) markers for human gene mapping. AB - A large collection of good genetic markers is needed to map the genes that cause human genetic diseases. Although nearly 400 polymorphic DNA markers for human chromosomes have been described, the majority have only two alleles and are thus uninformative for analysis of genetic linkage in many families. A few known marker systems, however, detect loci that respond to restriction enzyme cleavage by producing a fragment that can have many different lengths. This polymorphism is due to variation in the number of tandem repeats of a short DNA sequence. Because most individuals will be heterozygous at such loci, these markers will provide linkage information in almost all families. Ten oligomeric sequences derived from the tandem repeat regions of the myoglobin gene, the zeta-globin pseudogene, the insulin gene, and the X-gene region of hepatitis B virus, were used to develop a series of single-copy probes. These probes revealed new, highly polymorphic genetic loci whose allele sizes reflected variation in the number of tandem repeats. PMID- 3029875 TI - Ubiquitin is a component of paired helical filaments in Alzheimer's disease. AB - Paired helical filaments (PHF), which constitute a distinct type of pathological neuronal fiber, are the principal constituent of neurofibrillary tangles that occur in the brain of patients with Alzheimer's disease. Their insolubility in sodium dodecyl sulfate and urea has prevented the analysis of their subunit composition by gel electrophoresis. A monoclonal antibody (DF2) was isolated that specifically labeled PHF at both the light and electron microscopic levels. It labeled a small polypeptide (5 kilodaltons) that was shown to be ubiquitin in immunoblots of the soluble fraction of brain homogenates. To obtain direct evidence that ubiquitin is a component of PHF, PHF were treated with concentrated formic acid and digested with lysylendopeptidase; ubiquitin-derived peptides were then identified by reversed-phase high-performance liquid chromatography. Two fragments in the PHF digest were identified as derived from ubiquitin by protein sequencing. This procedure should make possible definitive identification of other PHF components. PMID- 3029874 TI - Heritable somatic excision of a Drosophila transposon. AB - A mutation in the white gene of Drosophila mauritiana that results from insertion of the transposable element mariner is genetically unstable in both germ cells and somatic cells. Somatic instability is indicated by the occurrence of animals having mosaic eyes with patches of pigmented cells on a peach-colored background. Normally uncommon, the frequency of mosaicism is so greatly enhanced in a particular mutant strain that virtually every animal in the strain is an eye color mosaic. The molecular basis of the mosaicism is the excision of the mariner element from its location in the DNA of the white gene in somatic cells. The phenomenon results from a single dominant genetic factor located in chromosome 3. Genetic control over the excision of transposable elements may play a role in determining the persistence of transposable elements in the genome. PMID- 3029876 TI - A new probe for the diagnosis of myotonic muscular dystrophy. AB - Myotonic muscular dystrophy (DM) is the most common muscular dystrophy, affecting adults as well as children. It is inherited as an autosomal dominant trait and is characterized by variable expressivity and late age-of-onset. Linkage studies have established the locus on chromosome 19. In order to identify tightly linked probes for diagnosis as well as to define in detail the DM gene region, chromosome 19 libraries were constructed and screened for restriction fragment length polymorphisms tightly linked to DM. A genomic clone, LDR152 (D19S19), was isolated that is tightly linked to DM; recombination fraction = 0.0 (95% confidence limits 0.0-0.03); lod score, 15.4. PMID- 3029878 TI - Rumen function with special reference to fibre digestion. PMID- 3029877 TI - Case report 408: Malignant fibrous histiocytoma of innominate bones and femur (multicentric). PMID- 3029879 TI - Bovine leukosis: an example of poor disease monitoring of international livestock shipments to developing countries. AB - Over the past few years, cattle imported into a number of countries have been shown to carry antibodies against bovine leukosis virus (BLV), the cause of enzootic bovine leukosis. The disease, imported into England via shipments of Canadian dairy cattle in the late 1970s, is still a significant problem in the United Kingdom. Many countries now require a blood test for serum antibodies to BLV as a precondition of shipment from certain endemic countries. Those cattle positive for BLV are excluded from shipment. Well-documented cases of shipments of hundreds of Canadian cattle without testing for BLV has been obtained. These shipments were to the African country of Malawi where the BLV status is unknown. Also, although the United States is endemic for BLV, most of the recent live cattle exports were received by countries which did not require a blood test for BLV. Of the 44 countries that imported U.S. cattle during 1982 and 1983, only 13 required BLV antibody testing before shipment. This report concerns one disease in one species. The millions of livestock shipped yearly between the countries of the world may harbor many viruses at risk of being introduced into susceptible animal populations. While international animal health agencies are doing a credible job of providing disease surveillance to member countries, there still needs to be improvements in disease information dissemination. PMID- 3029880 TI - [Problems of treating anorexia nervosa. Hospitalization, family relations, after care]. PMID- 3029881 TI - A personal view of advances in biliary tract surgery. PMID- 3029883 TI - Pancreatic islet cell tumors. PMID- 3029882 TI - Contralateral intracerebral hemorrhage as a complication of tumor biopsy. AB - The authors report two cases of contralateral intratumoral hemorrhage after tumor biopsy in patients with bilateral glioblastoma multiforme. There was no evidence of systemic hypertension during either procedure. The occurrence of hemorrhage distant to the site of biopsy may be related to some aspect of the surgical procedure and should be recognized as a possible complication of brain tumor biopsy procedures. PMID- 3029884 TI - Dentifrice abrasivity. The use of laser light for determination of the abrasive properties of different silicas. An in vitro study. AB - The abrasive properties of five different silicon dioxides were investigated using laser reflexion. The substances were milled, amorphous silica gels of which four were commercially available dentifrice ingredients (Syloblanc) and one an experimental product. The abrasion process was carried out on acrylic plates in a brushing machine using two series of slurries of the substances, with and without sodium lauryl sulphate. Significant differences in abrasivity of the substances were observed. When sodium lauryl sulphate was included, the rate of abrasion decreased for the Syloblanc substances but increased for the experimental substance. The contrast was primarily explained by the capability of the different substances to form stable foams owing to their thickening properties. The investigation confirmed that silicon dioxides exhibit a wide range of abrasivity and proved that sodium lauryl sulphate influenced the abrasion process. PMID- 3029885 TI - [Stable tachycardia as a cause of heart failure]. AB - The authors described a patient in whom the right vagus had been cut during removal of hormonally inactive chemodectoma in the bifurcation of the external and internal carotid arteries. Stable sinus and then atrial tachycardia (140-180 strokes per min.) developed on the operating table. Congestive cardiac insufficiency developed in 3 yrs. Treatment with diuretics cardiac glycosides was ineffective. Clinical and x-ray signs of cardiac insufficiency disappeared after atrial tachycardia was converted into atrial fibrillation with ventricular contractions up to 80 per min. A conclusion has been made that stable tachycardia causes the development of cardiac insufficiency. PMID- 3029886 TI - [Tissue respiration in chronic bronchitis]. AB - Altogether 445 patients with chronic bronchitis were under observation. Changes in tissue respiration were monitored by shifts in the concentration of pyruvic and lactic acids, LDH and cytochrome c oxidase activity and changes in the concentration of indices of the thiol spectrum. The results obtained showed that chronic bronchitis during exacerbation was characterized by disorders of tissue respiration which did not return to normal even in the course of therapy; aerobic as well as anaerobic oxidation was disturbed. In 91 patients 10-year shifts in indices were detected, however they were less than during exacerbation. Disorders of tissue respiration can be caused by two factors: inflammatory intoxication of organs and tissues and chronic oxygen insufficiency in tissues. Drugs promoting normal tissue respiration are recommended for pathogenetic treatment of chronic bronchitis. PMID- 3029888 TI - [Dietary treatment of colonic diseases]. PMID- 3029887 TI - [Effect of an eicosapentaenoic acid-rich diet on the arterial pressure and lipid level in the blood of hypertension patients]. PMID- 3029889 TI - [Dietary treatment of cancer]. PMID- 3029890 TI - [Angiotensin-converting enzyme inhibitor in heart failure]. PMID- 3029891 TI - [Canine parvovirus infection in dogs: an observation]. PMID- 3029893 TI - [Vaccination schedule for Parvovirus]. PMID- 3029892 TI - [The effect of vitamin administration to sows during weaning on the weaning estrus interval and on the number of live piglets in the next litter]. AB - The effect of injection of additional vitamins in sows during weaning on the weaning-oestrus interval and on the number of piglets born alive in the next litter was studied in 735 sows. On five commercial farms, a total of 735 sows were divided into 3 groups. The first group was injected with 10 ml of a preparation of AD3E. The second group was injected with 10 ml multivitamin. The third group was not treated. It is concluded that injection of additional vitamins did not have any effect on the weaning-oestrus interval and on the number of piglets born alive in the next litter. PMID- 3029894 TI - HLA and acute arthritis following human parvovirus infection. AB - HLA-DR4 frequency was raised in patients with persistent acute arthritis following human parvovirus infection, suggesting that DR4 positive individuals are more susceptible to develop joint complications following human parvovirus infection. PMID- 3029895 TI - Cardiac abnormalities in rats treated with methylphosphonothiolate. AB - Cardiac toxicity of methylphosphonothiolate (MPT), an organophosphorus compound, has been investigated in the rat. Subcutaneous injection of MPT (12 micrograms/kg body wt) induced cardiac arrhythmias, the occurrence of which was significantly more frequent than in the control group. Death rate among MPT-treated animals appeared to be in relationship with cardiac arrhythmias. Plasma nonesterified fatty acid concentrations increased in MPT-poisoned rats. cAMP and cGMP contents in myocardial tissue were unchanged 150 min after MPT administration, as compared with control. Similarly, no change has occurred in high energy compound levels. PMID- 3029896 TI - The pulmonary toxicity of talc and granite dust as estimated from an in vivo hamster bioassay. AB - A short-term animal bioassay was used to assess the toxicity of occupational dusts. We quantified pulmonary responses in hamsters exposed to granite (12% quartz) and talc (quartz and asbestos-free) dust collected from worksites. Personal samples collected on workers showed similar quartz content and particle size distributions to the high-volume samples collected for bioassays, thus demonstrating that the particulates were representative of worker exposure. We measured biochemical and cellular indicators of injury in bronchoalveolar lavage fluid (BAL) of animals exposed to dust suspensions by intra-tracheal instillation. The assays measured release of cytoplasmic and lysosomal enzymes into the cell-free supernatant of BAL; levels of albumin and red blood cells; changes in macrophage and polymorphonuclear neutrophil cell numbers; and in situ macrophage phagocytosis. Dose-response (0.15, 0.75, and 3.75 mg/100 g body wt) and time-course (1-14 days postexposure) studies were performed. One day after exposure, both talc and granite dust resulted in elevated enzyme levels, pulmonary edema, and increased cell numbers in BAL. Macrophage phagocytosis was also inhibited. Based on earlier studies, response levels were either intermediate between nontoxic iron oxide and toxic alpha-quartz or comparable with alpha-quartz. The response to granite dust diminished fairly rapidly over time. By contrast, after talc exposure, there was a more persistent elevation in enzyme levels, and macrophage phagocytosis remained depressed. These results indicate that, when a similar mass was deposited in the lungs, talc caused more lung injury than did granite. Better estimates of exposure-dose relationships in talc and granite workers as well as longer-term animal studies are required to evaluate the harmfulness of these work environments at present-day exposure levels. PMID- 3029897 TI - Mechanisms mediating cephaloridine inhibition of renal gluconeogenesis. AB - Incubation of renal cortical slices with cephaloridine (CPH) markedly inhibits pyruvate-supported gluconeogenesis, an effect which is independent of CPH-induced lipid peroxidation. CPH was found to inhibit pyruvate-supported gluconeogenesis in a time-and concentration-dependent manner. Pyruvate-supported gluconeogenesis was inhibited as early as 10 min following incubation of renal cortical slices with 5 mM CPH. Similarly, endogenous gluconeogenesis was impaired following CPH treatment. CPH depressed the renal cortical slice content of ATP by 50%, but only following 90 and 120 min of drug exposure, suggesting that mitochondrial dysfunction does not mediate the inhibition of gluconeogenesis by CPH. To identify the intracellular site(s) of CPH inhibition of gluconeogenesis, the effects of CPH on glucose production were evaluated using substrates catalyzed by rate-limiting reactions. CPH inhibited renal cortical slice gluconeogenesis when the following substrates were used: pyruvate (mitochondrial), oxaloacetate and fructose-1,6-diphosphate (FDP) (postmitochondrial), and glucose-6-phosphate (G6P, endoplasmic reticulum). Inhibition of G6P-supported gluconeogenesis occurred within 5 min of incubation with 5 mM CPH. Direct addition of CPH to microsomal suspensions inhibited G6Pase activity in a concentration-dependent fashion. By contrast, addition of CPH to cytosolic fractions did not affect FDPase activity. CPH increased the Km and decreased the Vmax of G6Pase, indicating mixed competitive and noncompetitive inhibition. These data indicate that the profound inhibition of renal cortical slice gluconeogenesis by CPH is mediated by inhibition of microsomal G6Pase activity. PMID- 3029898 TI - Effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin on hepatic and uterine estrogen receptor levels in rats. AB - Administration of a single dose of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD, 20 or 80 micrograms/kg) resulted in significantly decreased hepatic and uterine estrogen receptor (ER) levels in 25-day-old female Long-Evans rats. By contrast, estradiol (5 and 15 micrograms/kg) administration increased hepatic and uterine ER levels, while a combination of 2,3,7,8-TCDD plus estradiol resulted in uterine and hepatic ER levels which were similar or lower than those observed after treatment with only 2,3,7,8-TCDD. In addition, 2,3,7,8-TCDD significantly decreased the effects of estradiol on uterine wet weight increase. Competitive binding studies showed that estradiol did not bind to the aryl hydrocarbon (Ah) receptor and that 2,3,7,8-TCDD did not bind to the ER. The effects of structure on the activity of polychlorinated dibenzo-p-dioxin congeners on their activity to down-regulate hepatic and uterine ER levels were determined by using 2,3,7,8 TCDD, 1,2,3,7,8-pentachlorodibenzo-p-dioxin (PeCDD), 1,3,7,8-TCDD, and 1,2,4,7,8 PeCDD. Both 2,3,7,8-TCDD and 1,2,3,7,8-PeCDD exhibit high affinities for the Ah receptor and at dose levels of 80 micrograms/kg the hepatic ER levels were decreased 42 and 41%, respectively, and uterine ER levels were decreased 53 and 49%, respectively. 1,3,7,8-TCDD and 1,2,4,7,8-PeCDD bind less avidly to the Ah receptor and at dose levels of 400 micrograms/kg these compounds decreased hepatic ER levels 36 and 40%, respectively, and uterine ER levels 21 and 24%, respectively. These results support a role for the Ah receptor in the down regulation of uterine and hepatic ER levels in the female rat by 2,3,7,8-TCDD and this effect may be associated with the decrease in spontaneous mammary and uterine tumors observed in female rats treated with 2,3,7,8-TCDD. PMID- 3029899 TI - In vitro effects of 2,5 hexanedione on a melanoma cell line: a morphological study. AB - The effect of 2,5 hexanedione (2,5 HD) on a cultured human melanoma cell line (JR8) was explored. The addition of the toxicant at noncytolitic concentrations (0.08-0.16%) to the monolayers for 24 and 48 h, resulted in an irreversible inhibition of cell proliferation. Cessation of melanoma cell proliferation was accompanied by wide changes in morphological features of cells still adhering to the substrate. Incubation with the toxicant seemed to induce a differentiative process characterized mainly by a significant increase in cell protrusions. Melanoma cells, losing their bipolar appearance, often increased cell size and developed long dendritic and axon-like processes sometimes ramified in distal portions. Electron microscopic observations established that a change in the polarized appearance of control cells often occurred with 2,5 HD treatment and that a regular arrangement of organelles and cytoskeletal elements was detectable within these dendritic and axon-like protrusions. Furthermore, immunocytochemical studies confirmed an involvement of microtubules and actin network within cell prolongations. After the differentiative process a necrotizing effect occurred, inducing a progressive loss of viable, dendritic cells after 4 or 5 days. Incubation with cyclic AMP was ineffective in control cells while after 2,5 HD treatment seemed to increase the survival rate of neuronal-like cells. Possible mechanisms for the growth inhibitory and differentiative effects of 2,5 HD were discussed. PMID- 3029900 TI - Alkaline phosphodiesterase I release from eucaryotic plasma membranes by phosphatidylinositol-specific phospholipase C. II. The release from brush border membranes of porcine intestine. AB - Ectoenzyme release from porcine intestinal brush border membranes by phosphatidylinositol-specific phospholipase C of Bacillus thuringiensis was studied. Alkaline phosphodiesterase I, alkaline phosphatase and 5'-nucleotidase were released from both slices and brush border membranes. The pattern of alkaline phosphodiesterase I release was the same as that of alkaline phosphatase. The release of alkaline phosphodiesterase I induced by phospholipase C was dependent on, or proportional to, the reaction time and the concentration of phospholipase C. The Arrhenius plot for phosphodiesterase I release showed a single break at 30 degrees C for brush border membranes. Only 40% of alkaline phosphodiesterase I present in the brush border membranes were solubilized by phosphatidylinositol-specific phospholipase C treatment. The data indicate the presence of two forms of phosphodiesterase I, which are different in their sensitivity to phospholipase C. The released alkaline phosphodiesterase I had a molecular weight of 240,000 and was activated by Mg2+ and Ca2+, but strongly inhibited by EDTA. PMID- 3029902 TI - Immunotoxicology studies on octoxynol-9 and nonoxynol-9 in mice. AB - In a double-blind study, mice were injected intraperitoneally with 0.2 ml 0.2% octoxynol-9 (O-9), 0.2 ml 0.2% nonoxynol-9 (N-9), or 0.2 ml saline (control) daily for 24 days. Another control group received no treatment. All mice were immunized twice with sheep red blood cells (SRBC) and bled by caudal incision. Mice receiving N-9 lost weight (P less than 0.02), had smaller livers (P less than 0.05), and showed enlarged spleens (P less than 0.05). The N-9-treated mice did not differ from either control group in the primary or secondary anti-SRBC responses, leukocyte (WBC) counts, or in the sizes of the kidneys, hearts, lungs, or thymuses. Mice receiving O-9 showed no significant differences from either control group in any of these tests. Serum immunoglobulin M (IgM) and immunoglobulin G (IgG) levels were similar in mice treated with O-9, N-9, or saline. All 3 groups had higher levels of both classes of immunoglobulins on day 16 than did untreated controls. This study shows that O-9, given to mice in doses 3 times that used by humans, is nontoxic, whereas the same dose of N-9 has minor deleterious effects. PMID- 3029901 TI - Pyrethroid insecticides and radioligand displacement from the GABA receptor chloride ionophore complex. AB - Radioligand binding displacement studies were conducted to determine the effects of Type I and II pyrethroids on [3H]flunitrazepam (FLU), [3H]muscimol (MUS), and [35S]t-butylbicyclophosphorothionate (TBPS) binding. Competition experiments with [3H]FLU and [3H]MUS indicate a lack of competition for binding by the pyrethroids. Type I pyrethroids failed to compete for the binding of [35S]TBPS at concentrations as high as 50 microM. Type II pyrethroids inhibited [35S]TBPS binding to rat brain synaptosomes with Ki values ranging from 5-10 microM. The data presented here suggest that the interaction of Type II pyrethroids with the gamma-aminobutyric acid (GABA) receptor-ionophore complex is restricted to a site near the TBPS/picrotoxinin binding site. PMID- 3029903 TI - Prenatal exposure to methylmercury alters development of adrenergic receptor binding sites in peripheral sympathetic target tissues. AB - In order to assess the impact of prenatal exposure to methylmercury on sympathetic neurotransmission, effects on development of adrenergic receptor binding sites in peripheral tissues were evaluated. In the liver, methylmercury produced a dose-dependent increase in alpha 1-, alpha 2- and beta-receptor binding of radioligands throughout the first 5 weeks of postnatal life. Similarly, renal alpha-receptor subtypes showed increased binding capabilities, but binding to beta-receptor sites was reduced. At least some of the changes in receptors appear to be of functional significance, as physiological reactivity to adrenergic stimulation is altered in the same directions in these two tissues. The actions of methylmercury displayed tissue specificity in that the same receptor populations were largely unaffected in other tissues (lung, heart). These results suggest that methylmercury exposure in utero alters adrenergic responses through targeted effects on postsynaptic receptor populations in specific tissues. PMID- 3029905 TI - Evaluation of a confidential method of excluding blood donors exposed to human immunodeficiency virus: studies on hepatitis and cytomegalovirus markers. AB - A confidential self-administered questionnaire was given to all blood donors prior to donation (n = 95,917). The questionnaire describes groups at increased risk of acquired immunodeficiency syndrome (AIDS) and requires the donor to designate his blood either for laboratory purposes or for transfusion. In a previous communication, we reported that donors in the former group had a much higher prevalence of antibody to human immunodeficiency virus (HIV) than age, sex and clinic matched controls or a group of "miscellaneous" donors who did not fill out the form properly. In this communication, we report results of tests for other viral markers performed on the three designation groups, namely laboratory designated, miscellaneous and controls. We found that the former two groups had a higher prevalence of antibody to hepatitis B surface antigen (anti-HBs), hepatitis B core antigen (anti-HBc) and cytomegalovirus (anti-CMV) than controls, but there were no differences in alanine aminotransferase (ALT) levels among the groups. In addition, the laboratory-designated group had a higher prevalence of hepatitis B surface antigen (HBsAg) than the general donor population. These data indicate that a questionnaire designed to ascertain AIDS high-risk donors is valuable in excluding donors who may be carriers of other viruses as well. PMID- 3029904 TI - Look-back: preliminary experience of AABB members. American Association of Blood Banks. AB - By mid-fall of 1986 the "look-back" process conducted by blood banks belonging to the American Association of Blood Banks was about 50 percent complete. Over one half of the traced recipients had died; 15 percent were alive and negative for antibodies to the human immunodeficiency virus and 16 percent alive but had seroconverted. In a small number of cases physicians or patients elected not to be tested. No major problems were reported. PMID- 3029906 TI - Studies of the sensitivity and reproducibility of commercial kits to detect antibodies to the human immunodeficiency virus. AB - Nine serial three-fold dilutions (1:1 to 1:6561) were prepared from 18 sera obtained from hemophiliacs confirmed to have antibodies to the human immunodeficiency virus. The dilutions were tested with five different commercial enzyme immunoassay kits and twelve sera were retested 5 to 7 months later by different lots of three kits. The dilution that gave an absorbance (OD) equal to the cut-off OD was considered as the titer of antibody. Sensitivities of the kits were compared by statistical and regression analysis; the same approach was used for studying reproducibility from the results of retesting. The highest titers were found with the Wellcome kit, the lowest with Organon and Pasteur kits, whereas intermediate values were found with the Sorin/Biomedica and Electronucleonics kits. With the Organon and Wellcome kits, excellent reproducibility was observed on later retesting; however, a significant change in titers was seen on retesting the Sorin kit. PMID- 3029907 TI - Prevalence of human immunodeficiency virus antibodies in northern Nigerian blood donors. PMID- 3029908 TI - Variables predicting bacterial and fungal infections after allogeneic marrow engraftment. AB - Sixty-seven consecutive patients with aplastic anemia or leukemia who had been treated by allogeneic marrow transplantation and had survived for more than 1 month were surveyed in order to determine the incidence of nonviral infections occurring from 1 month to 3 years after transplantation. Twenty-eight of the 67 patients had one or more infections during this period. Around 20% suffered from pulmonary infections and 20% were classified as having a systemic infection. Ten patients died of bacterial or fungal infection, of whom 6 had graft-versus-host disease. In multivariate analyses acute graft-versus-host disease (P less than 0.0009), splenectomy (P less than 0.02), cytomegalovirus infection (P less than 0.05), and a low marrow cell dose (P less than 0.07) were correlated with nonviral infections. PMID- 3029910 TI - Metabolic activity of the transplanted liver. PMID- 3029909 TI - T cell subset patterns in cyclosporine-treated renal transplant recipients with primary cytomegalovirus disease. PMID- 3029911 TI - Cyclosporine A fails to alter innate host resistance to murine hepatitis virus strain 3 infection in A/J mice. PMID- 3029912 TI - A simple methodological principle for large scale extraction and purification of collagenase-digested islets. PMID- 3029913 TI - Isolation of viable islets of Langerhans from collagenase-perfused canine and human pancreata. PMID- 3029915 TI - Morbid outcome of cytomegalovirus-negative transplant recipients receiving cytomegalovirus-positive kidneys. PMID- 3029914 TI - Effect of initial versus delayed cyclosporine therapy in cadaveric renal transplant patients. PMID- 3029916 TI - Viral infections in kidney transplant patients immunosuppressed with cyclosporine. PMID- 3029917 TI - Criteria of selection for liver transplantation. PMID- 3029918 TI - Incidence of cytomegalovirus infection and its relationship to donor-recipient serologic status in liver transplantation. PMID- 3029919 TI - Bronchoalveolar macrophage-lymphocyte reactivity in heart-lung transplant recipients. PMID- 3029920 TI - Autotransplantation of the adrenal gland using microvascular anastomosis. PMID- 3029922 TI - Urinary neopterin in bone marrow recipients. PMID- 3029921 TI - Risk factors for interstitial pneumonitis following allogeneic bone marrow transplantation for severe aplastic anemia: a preliminary report. PMID- 3029923 TI - Cell-mediated immune responses in rat cytomegalovirus infection. PMID- 3029924 TI - Some aspects of iodine deficiency in Nepal. PMID- 3029926 TI - Bilateral rhabdomyomatous tumor relapsed as typical triphasic Wilms' tumor. AB - The authors report on a child affected with bilateral renal tumor, which was treated with cancer chemotherapy before and after surgery. Twenty-eight months after the discontinuance of therapy, a neoplasm was disclosed in the left kidney and then removed. Histologically, the bilateral tumor excised by the first surgery could be classified as biphasic Wilms' tumor, rhabdomyomatous variant, whereas the neoplasm removed by the second surgery was the typical triphasic Wilms' tumor. The authors suggest that preoperative chemotherapy might have played a role in the histologic changes of the initial tumor. Nonetheless, it is also tempting to postulate that the two histologic variants of Wilms' tumor could have occurred in the patient in spite of any treatment. PMID- 3029925 TI - [Transformation of bone marrow cells in rodents by recombinant plasmid pBRSV]. AB - Bone marrow cells (mouse strain CBA/Ca and Syrian hamster cells) were transformed with pBRSV DNA containing T-antigen of the SV40 virus. The SV40 T-antigen in transformed cell was detected in 0.5% cases by immunofluorescence with specific antibodies. Extrachromosomal localization of recombinant DNA was shown by means of retransformation of E. coli cells with cytoplasmic spleen DNA from mice previously injected intravenously the transformed bone marrow cells. PMID- 3029927 TI - [The skin and colonic polyposis]. PMID- 3029928 TI - [Survival in primary bronchial cancer]. PMID- 3029929 TI - [A rare malignant pulmonary tumor: pneumoblastoma. Apropos of a case report and review of the literature]. PMID- 3029931 TI - Dedifferentiated chondrosarcoma: an ultrastructural study of two cases, with immunocytochemical correlations. AB - The clinicopathologic features of two cases of dedifferentiated chondrosarcoma (DCS) are presented, in which anaplastic components showed the electron microscopic features of malignant fibrous histiocytoma, as well as immunoreactivity for alpha-1-antitrypsin and alpha-1-antichymotrypsin. Rare cells also displayed S100 protein in high-grade areas of the primary tumors, but a pulmonary metastasis lacked this determinant. These findings could be interpreted as reflecting a retained potential for primitive chondrogenesis in primary DCS, which may be lost in its metastases. In all other respects, this tumor appears to assume the morphological and immunocytochemical attributes of a fibrohistiocytic neoplasm. PMID- 3029933 TI - Limitations of the usefulness of microvillous ultrastructure in distinguishing between carcinoma primary in and metastatic to the lung. AB - We performed ultrastructural analysis on 70 consecutive patients with solitary cancers in lung with the following histologic classifications: adenocarcinoma (42 cases), bronchioloalveolar carcinoma (13), large cell carcinoma (4), and adenosquamous carcinoma (11). Of these 70 cases, nineteen (13 adenocarcinomas, 4 bronchioloalveolar carcinomas, and 2 adenosquamous carcinomas) contained cell surface microvilli with microvillous core rootlets and/or glycocalyceal bodies. Subsequent clinical followup revealed that three of these 19 cases were actually metastatic colon carcinoma. The remaining 16 patients are currently free of extrathoracic primary disease and are therefore, presumably, primary carcinoma of the lung. Since both primary and metastatic tumors showed cell surfaces with microvilli having core rootlets and glycocalyceal bodies, we conclude that the presence of these ultrastructural features does not always permit the distinction between primary and metastatic adenocarcinoma in lung. PMID- 3029930 TI - [Vitamin-resistant rickets preceding Wilson's disease]. PMID- 3029932 TI - Fibrolamellar carcinoma of liver: a primary malignant oncocytic carcinoid? AB - Immunohistochemical and ultrastructural findings in two cases of fibrolamellar carcinoma of the liver and two cases of hepatocellular carcinoma of the common histologic type are described. Ultrastructural examination of both cases of fibrolamellar carcinoma revealed the presence of neurosecretory (NS) granules which were sparse in some cells and abundant in others. Many of the tumor cells had a distinct oncocytic appearance with abundant mitochondria. A portion of the glutaraldehyde-fixed neoplasm was processed for the uranaffin reaction (an ultrastructural cytochemical stain specific for the NS granules of neuroendocrine tissue). Abundant uranaffin-positive granules were found in the neoplastic cells of both cases of fibrolamellar carcinoma, whereas no uranaffin-positive granules were found in hepatocellular carcinoma of the common histologic type. There was no statistical difference in the mean diameter of the uranaffin-positive granules measured from both cases. Immunohistochemistry revealed the presence of neuron specific enolase (NSE) and serotonin in one of the two cases of fibrolamellar carcinoma and no NSE staining in two cases of hepatocellular carcinoma of the common histologic type. These findings suggest that some liver tumors presenting histologically as fibrolamellar carcinoma may be neuroendocrine in nature. PMID- 3029934 TI - [IgM M-component associated neuropathy. A new autoimmune disease entity]. PMID- 3029935 TI - [A new polyvalent vaccine against enteral infections in newborn calves]. AB - The most frequent microbial causative agents of massive diarrheas in new-born calves kept on large cattle farms in the CSSR are rotaviruses, coronaviruses and enterotoxigenic strains of E. coli, manifesting themselves as complicated virus bacterial infections. An inactivated polyvalent adjuvant vaccine has been developed for the prevention and specific prophylaxis of these enteral infections; the vaccine contains bovine rotavirus, bovine coronavirus and three enterotoxigenic serotypes of E. coli with protective antigen K 99. The rotavirus and coronavirus are propagated on the stable cellular line MDBK and inactivated with 0.2% formalin, the Escherichia strains are submersed in the MINCA culture medium during their cultivation and inactivated with 0.5% formalin. The vaccine was prepared as a blend of the same amounts of rotavirus and coronavirus and of such an amount of bacterin so that 1 ml of the vaccine will contain 10(9) bacteria. One part of oil adjuvant was added to five parts of the virus-bacterial blend and the blend was homogenized in the Ultraturax apparatus. The vaccine is to be used for immunization of pregnant cows and heifers; in these animals it induces the production of specific antibodies to all antigens contained in the vaccine. Its immunogenic effects were checked in 32 calves and 38 cows in the herds with the occurrence of diarrheas caused by both enteropathogenic viruses and enterotoxigenic escherichia. It was demonstrated that the inactivation did not influence in either of the viruses the process of inducing the production of specific antibodies, and the antibody response of the calves and heifers after application of 2 ml of complete inactivated vaccine was equally strong as after application of live vaccine containing only rotavirus and coronavirus. The level of the rotavirus antibodies increased on the average 30 times and 200 times, coronavirus antibodies twice and four times. The antibody response to coronavirus was negatively influenced by the relatively high levels of antibodies before vaccination. The antibody response to antigen K 99 was expressive in all cases. PMID- 3029936 TI - Feline leukemia virus-associated enteritis--a condition with features of feline panleukopenia. AB - Infection with feline leukemia virus (FeLV) was demonstrated immunohistologically in 218 necropsied cats suffering from enteritis. The animals were divided into three groups according to histopathological criteria. The first group exhibited the signs of feline panleukopenia in intestine, lymphoid tissues, and bone marrow. Only 1.6% of these animals were FeLV-infected. The animals of the second group had histopathological alterations as seen in cats suffering from feline panleukopenia, but these were found only in the intestine and not in lymphoid tissues or bone marrow. Of these 71.9% were infected with FeLV. The third group consisted of all other cats suffering from enteritis of which 6.3% were FeLV positive. The association between FeLV infection and the lesions seen in the animals of group 1 (feline panleukopenia) and group 3 (other types of enteritis) is statistically not significant whereas the alterations exhibited by the cats of group 2 are significantly FeLV-associated. Cats with FeLV-associated enteritis (group 2) are of a mean age of about 2.5 years and are significantly older than animals with feline panleukopenia which are of a mean age of about half a year. Thus a FeLV-associated enteritis exists as a histopathologically recognizable condition which sometimes might be mistaken for feline panleukopenia in routine post-mortem investigations. PMID- 3029938 TI - A canine ovarian germ cell tumor with extraembryonic differentiation. PMID- 3029937 TI - Peripheral neuropathy associated with malignant neoplasms in dogs. AB - Common peroneal and ulnar nerves of 21 dogs between 2 and 15 years of age with malignant tumors were examined for possible paraneoplastic effects. Percent of abnormalities in single-teased fiber studies exceeded the confidence limits (P = 0.99) established for age-matched control dogs in 16 of 21 dogs (76%) with malignancies, none of which manifested clinical signs of polyneuropathy. The incidence of abnormalities was higher in common peroneal nerve (73%) than in ulnar nerve (57%). Lesions were characterized by a mixture of demyelination/remyelination and axonal degeneration; however, the former lesions predominated in most affected dogs. The severity of the neuropathy varied with different tumors, with the most severe lesions observed in a bronchogenic carcinoma (59%), a mammary adenocarcinoma (59%), a malignant melanoma (48%), and an insulinoma (47%). PMID- 3029939 TI - A study of 31 cases of gastric carcinoma in dogs. AB - Over a five year period 31 dogs were diagnosed as having advanced gastric carcinoma. The most frequent clinical features were vomiting, polydipsia and weight loss. A predisposition to the tumour was found in the rough collie and Staffordshire bull terrier. In 18 dogs the main finding endoscopically was a large deep ulcer with thickened, irregular rims and walls. Fluoroscopically a marked irregularity of the mucosal surface was noted in 10 dogs. Pathologically, large ulcers with thickening of the stomach wall and involvement of the serosal lymphatics were evident in 17 dogs, and similar ulcers with involvement of the gastric lymph nodes were evident in 18 dogs. PMID- 3029940 TI - Prevalence of IBV antibodies in turkey breeding flocks in Israel. PMID- 3029941 TI - Lymphocyte 5'-nucleotidase activities in normal dogs and in dogs with malignant lymphoma. AB - Plasma membrane 5'-nucleotidase (5'NT) activity was assayed in lymph node lymphocytes from seven normal control dogs and in malignant lymphocytes from 25 dogs with lymphoma. The lymphoid tumors were classified according to the NCI Working Formulation into five histologic subtypes. When compared with normal controls significantly lower 5'NT activities were found in the lymphoblastic, small lymphocytic and diffuse large (noncleaved) subtypes while no significant differences were observed in the immunoblastic or small noncleaved groups. In addition, dogs with hypercalcemia or paraproteinemia had decreased 5'NT levels. However, no significant differences were found between prognostic groups or histologic subtypes of the NCI classification. In conclusion, canine lymphoma subtypes had generally decreased 5'NT activities which appeared to reflect the B or T cell lineage, degree of maturation and enzyme activity of the cell of origin. PMID- 3029942 TI - Patterns of bovine T cell-mediated immune responses to bovine herpesvirus 1. AB - Lymphocyte proliferation was used to evaluate T cell-mediated immune responses to different isolates of Bovine Herpesvirus 1 (BHV-1). Groups of high, moderate, and low responses were observed when lymphocytes from three breeds of dairy cattle were stimulated with each of the BHV-1 isolates. Proliferation of cells in the low responding group could be augmented by exogenous IL-2. The mechanism of unresponsiveness by cells from one individual whose response was not altered by IL-2 supplementation was further investigated. The patterns of response by limiting dilution frequency analysis eliminated the possibility that this individual lacked responsive cells but suggested the presence of regulatory cell interactions which resulted in the observed low proliferative response. These results show that animals exposed to the same environment can vary greatly in their ability to respond to BHV-1. At least two mechanisms may be responsible for low proliferative responses in vitro: inadequate levels of IL-2 and the presence of suppressor cells. PMID- 3029944 TI - Natural cell-mediated cytotoxicity to cells infected with infectious bovine rhinotracheitis virus. AB - Cell-mediated cytotoxicity against viral-infected cells was demonstrated in a 6 hr 51Cr release assay. Peripheral blood mononuclear leukocytes from both infectious bovine rhinotracheitis virus (IBRV)-infected and noninfected cattle exhibited preferential lysis against IBRV-infected primary bovine embryonic kidney (BEK) cells compared to cells infected with pseudorabies virus and noninfected BEK cells. Addition of specific antibody to the assay did not enhance cytotoxicity. The effector cell was a nonadherent cell which was either spontaneously enriched or generated during in vitro cultivation. Maximal cytotoxic activity was detected in peripheral blood mononuclear cells cultured for 3 to 5 days. Several factors affected the magnitude of cytotoxicity during the assay: target cell type, concentration of viral inoculum, duration of effector and target cell contact. It is suggested that target cell lysis was a form of natural cell-mediated cytotoxicity mediated by a cell which has different characteristics from the typical human and murine NK cell. PMID- 3029943 TI - Serological diagnosis of influenza A infections in the horse by enzyme immunoassay. Comparison with the complement fixation test. AB - An enzyme immunoassay (EIA) using horseradish peroxidase and a type-specific antigen is described for the detection and quantitation of anti-influenza antibodies in the horse. Compared with the complement fixation (CF) test (using the same antigen), EIA proved to be superior with respect to sensitivity and reliability. The internal variation of EIA was low and thus small titres in EIA can be considered of diagnostic significance. However, no strict correlation with CF was observed. The use of an immunoconjugate against equine IgM in parallel with IgG would certainly improve the sensitivity of the test, especially in early stages of infection. PMID- 3029945 TI - Neutrophil function in sheep experimentally infected with bovine leukemia virus. AB - In vitro neutrophil adherence, random migration, chemotaxis, resting and phagocytosis-associated oxygen consumption and bactericidal responses were assessed in sheep experimentally infected with bovine leukemia virus (BLV). Neutrophil function was examined in two groups of 9 control and 9 BLV-infected sheep at 0, 1, 2, 3, 5, 7, 11 and 15 weeks post-infection. Enhanced neutrophil adherence, chemotaxis and resting oxygen consumption responses were found in the infected group at 2, 11 and 15 weeks respectively. Significant alterations between groups were not demonstrated during the other time intervals. PMID- 3029946 TI - Sarcomatoid carcinoma of the breast: an immunohistochemical study of six cases. AB - Six cases of sarcomatoid carcinoma of the breast (SCB) were studied with a panel of antibodies directed against epithelial and sarcomatoid components. The monoclonal antibodies (MoAb) AE-1/3, CAM 5.2, and CEA were used to detect epithelial differentiation; polyclonal antibodies against S-100 protein and MoAb against the intermediate filaments desmin and vimentin were used to detect mesenchymal differentiation in the sarcomatoid component. Six cases of invasive duct carcinoma (IDC) and two cases of cystosarcoma phyllodes (CP) were compared to SCB using the same panel of antibodies. In all three groups studied, the epithelial component in the majority of cases stained with anti-cytokeratin antibodies. S-100 protein antibodies stained the epithelial and sarcomatoid components in four cases of SCB; vimentin MoAb stained the epithelium in two cases and the sarcomatoid component in four cases of SCB, while MoAb CEA failed to stain any component of SCB. In contrast, the epithelium in five of six cases of IDC stained with CEA MoAb and only one of six stained for S-100 protein. Possible reasons for the discrepant immunohistochemical staining patterns among SCB, IDC and CP are discussed, in addition to the limitations and pitfalls of immunohistochemistry in diagnostic surgical pathology. PMID- 3029947 TI - [Detection of type 1 and 2 herpetic antigens by immunofluorescence in patients with chronic or recurrent keratoconjunctivitis receiving therapy based on etiology]. AB - Immunofluorescence (IF) techniques were used to evaluate the therapeutical efficiency of moroxidine hydrochloride against herpes virus (HSV) induced keratoconjunctivitis. In 56 out of the 77 followed-up and treated patients, the IF reactions revealed the presence of both types 1 and 2 of herpes virus, while in 21 only type 1 was found. The treatment with moroxidine hydrochloride led to a reduction of 63% and 92%, respectively, of the incidence of herpes virus type 1 and 2 antigens in the conjunctival cells, coincident with the improvement of clinical symptoms or persistent recovery. PMID- 3029948 TI - [Hemagglutination inhibition test for the diagnosis of coronavirus infection in cattle]. AB - The main aim of the work was to obtain a viral corpuscle suspension with high hemagglutinating activity to be used as an antigen in HAI tests. The bovine coronavirus SC-1 strain was cultivated on ovine foetal renal cells and gave a suspension with a hemagglutinating titer of 1:64-1:128. A monospecific anti coronavirus immune serum to be used in the HAI test, too, was prepared by hyperimmunization of a gnotobiotic calf. Its hemagglutinating and neutralizing titers were of 1:2560 and, respectively, 8.5 log. The HAI test was performed using the following reagents: the coronavirus suspension as a hemagglutinating antigen, a 1% suspension of mouse red blood corpuscles, the monospecific anti coronavirus immune serum as a positive control, and a negative serum. 265 serum samples were investigated, collected from cows and calves in 18 farms where neonatal gastroenteritis was diagnosed. In 72.4% out of these samples there were positive results in the IHA test. PMID- 3029949 TI - Considerations about the possible function of ceruloplasmin in influenza and parainfluenza virus infections. AB - The experimental data reveal that ceruloplasmin is a serum nonspecific factor acting during the early and late stages of infection with some respiratory viruses. This complex action seems to be based both on the enzymatic activity and on the copper-glycoprotein structure of ceruloplasmin. The changes in progen antigenicity produced by ceruloplasmin are probably involved in the mechanism of the antigenic shift and drift. PMID- 3029950 TI - The structure and cloning of orf virus DNA. AB - A map of cleavage sites for the restriction endonucleases EcoRI, HindIII, BamHI, HpaI, and KpnI for a New Zealand strain of orf virus (NZ2) DNA has been deduced. Also, the entire genome, apart from approximately 0.1 kbp at each end, has been cloned into various vectors. The genome is 139 kbp in length and, in common with other poxviruses, has inverted terminal repetitions and crosslinked ends. PMID- 3029951 TI - Association of bluetongue virus with the cytoskeleton. AB - Analysis of the distribution of [35S]methionine-labeled virus proteins following lysis of bluetongue virus (BTV)-infected cells with nonionic detergents showed that a major proportion of the virus-specific proteins was located in the insoluble nuclear-cytoskeletal fraction. Neither the proportion nor the species of virus protein associated with the cytoskeleton was altered following treatment of infected cells with microtubule or microfilament disrupting drugs (colchicine and cytochalasin B, respectively). Electron microscopic examination of BTV infected cells revealed cytoplasmic virus-specified tubules, viral inclusion bodies (VIB), and progeny virus particles. Whole-mount transmission electron microscopy of nonionic detergent-extracted cells demonstrated the association of VIB, virus particles, and tubules with the cytoskeleton. The identity of virus particles was confirmed with an immunogold labeling technique using a neutralizing monoclonal antibody to BTV protein VP2. Several lines of evidence indicate that virus particles, VIB, and tubules bind to intermediate filaments in BTV-infected cells. These structures remained cytoskeleton associated in infected cells treated with colchicine or cytochalasin B. Linear arrays of filament associated virus particles were formed around VIB following colchicine-induced juxtanuclear aggregation of intermediate filaments. Virus particles were associated with filaments approximately 10 nm in diameter. Filaments associated with virus particles reacted with an anti-vimentin monoclonal antibody in an immunogold labeling procedure. PMID- 3029952 TI - The amino terminus of the bacteriophage D108 transposase protein contains a two component sequence-specific DNA-binding domain. AB - We have cloned various amino-terminal domains of the transposable bacteriophage D108 A protein (transposase) into a high-copy-number expression vector and visualized the D108 polypeptides and fusion proteins expressed by the recombinant plasmids. By using crude protein extracts made from strains harboring these recombinant plasmids, we have performed band competition assays and DNasel footprinting on a 32P-end-labeled DNA restriction fragment which contained the Mu right end (to which the Mu A protein binds) and have shown that these fusion proteins in crude extracts can specifically bind to this DNA substrate. Furthermore, we have divided the amino-terminal 13 kDa of the D108 A protein (which may contain two bi-alpha-helical protein structures) in half and have shown that each half is capable of independent binding to the Mu attR site. These results suggest that the D108 transposase protein contains multiple DNA-binding domains which may be required for the complex protein-DNA interactions of D108 transposition. PMID- 3029954 TI - Two DNA antirestriction systems of bacteriophage P1, darA, and darB: characterization of darA- phages. AB - Bacteriophage P1 is only weakly restricted when it infects cells carrying type I restriction and modification systems even though DNA purified from P1 phage particles is a good substrate for type I restriction enzymes in vitro. Here we show that this protection against restriction is due to the products of two phage genes which we call darA and darB (dar for defense against restriction). Each of the dar gene products provides protection against a different subset of type I restriction systems. The darA and darB gene products are found in the phage head and protect any DNA packaged into a phage head, including transduced chromosomal markers, from restriction. The proteins must, therefore, be injected into recipient cells along with the DNA. The proteins act strictly in cis. For example, upon double infection of restricting cells with dar+ and dar- P1 phages, the dar+ genomes are protected from restriction while the dar- genomes are efficiently restricted. PMID- 3029953 TI - Conservation and variation in orf virus genomes. AB - The genomes of several orf virus strains were analyzed with the restriction endonucleases EcoRI, HindIII, BamHI, and KpnI, and cleavage site maps were deduced. In general, the right half of the genome showed conservation of restriction sites compared with the left half. Variations in size of up to 0.5 kbp were found within an inverted terminal repetition, and a 1-kbp deletion was detected in some strains in a subterminal fragment at the left end. A region of approximately 20 kbp, some 12 kbp in from the left end, showed the greatest cleavage site variability although there was no evidence of large deletions in this region. A 1.55-kbp cloned DNA fragment from the internal variable region of NZ2 failed to hybridize to the DNA from three other strains. A fragment in the variable region of strain NZ7 was cloned and compared by hybridization and restriction endonuclease analysis with cloned NZ2 fragments from the same region. The region of nonhomology extended for at least 2.75 kbp. It is suggested that this internal variable region may provide sites for the insertion of foreign genes. PMID- 3029955 TI - Expression and proteolytic processing of the darA antirestriction gene product of bacteriophage P1. AB - The darA gene coding for one of the two bacteriophage P1 antirestriction functions is expressed late after infection or induction. The protein is made as a high-molecular-weight soluble precursor. This is proteolytically cleaved to the mature form, which is a structural component of the phage head. Defective mutants of the phage have been found in which the synthesis of gpdarA is normal but processing does not take place. These mutations all map to the same region of the P1 genome and we propose that they lie in the structural gene for the processing protease. PMID- 3029956 TI - Isolation of a capsid protein of bluetongue virus that induces a protective immune response in sheep. AB - A method to purify the neutralization specific antigen of bluetongue virus P2 in large amounts has been developed. The purified protein is free from virus specified or cellular contaminants and its immunological specificity has been preserved. The purification is based on the observation that protein P2 can be dissociated from the virion by treatment with monovalent or divalent salts. The salt concentration required to solubilize the outer capsid proteins is pH dependent and in general decreases with a decrease in pH. P2 purified by extraction from polyacrylamide gels does not induce immune-precipitating or neutralizing antibodies. The response against P5, on the other hand, is much less conformational dependent and P5 purified from gels readily induces P5 precipitating antibodies in rabbits. These antibodies do not neutralize the virus. Purified P2, immunoabsorbed with anticore serum to remove trace amounts of P7, was injected into sheep. An initial dose of 50 micrograms of P2 was sufficient to induce P2-precipitating antibodies as well as neutralizing and hemagglutination-inhibiting antibodies. These sheep were fully protected against challenge with a virulent strain of the same BTV serotype. Lower doses of P2 still provided a significant level of protection even though no neutralizing antibodies could be detected. PMID- 3029957 TI - Uncoating of parental bluetongue virus to core and subcore particles in infected L cells. AB - A study was made of the fate of parental bluetongue virus (BTV) in infected cells. Within the first hour after infection, the BTV particles are converted to core particles with the loss of major capsid polypeptides P2 and P5. The particles are able to synthesize mRNA in vitro in a transcription reaction characterized by a temperature-dependent inhibition at high core concentrations. From about 6 hr after infection a second uncoating event is observed in which the 470 S core particles are converted to 390 S subcore particles. These particles are morphologically strikingly different from core particles and have a skeletonlike structure with a hexagonal profile and a side to side diameter of 40 nm. These subcore particles contain only one major structural protein, P3, and three minor proteins, P1, P4, and P6. They do, however, contain all 10 double stranded RNA segments. The results suggest that the characteristic capsomeres on the surface of core particles are composed mainly of P7, the soluble group specific antigen of BTV. The subcore particles are stable only at very low salt concentrations. Under these conditions no transcriptase activity can be demonstrated. PMID- 3029958 TI - In vitro assembly of bovine rotavirus nucleocapsid protein. AB - The nucleocapsid protein (VP6) of bovine rotavirus was purified from in vitro derived single shelled particles by CaCl2 or LiCl treatment. The protein exhibits polymorphism. Specifically, hexamers and small hexagonal lattices were present in many of the samples. Tubular particles formed between pH 5.0 and 9.0 were moderately stable to changes in temperature and ionic strength and were shown to be composed of nucleocapsid protein. Their formation is fully reversible. Spherical particles resembling single-shelled virus formed at pH 4.0. A novel structure in the form of sheets composed of a small-hole lattice formed in samples shifted from pH 6.0 to 4.0. The results demonstrate the importance of the nucleocapsid protein and of protein-protein interactions for rotavirus assembly. PMID- 3029959 TI - Cis-acting transcriptional regulatory sequences in the gibbon ape leukemia virus (GALV) long terminal repeat. AB - Gibbon ape leukemia viruses (GALV) are a group of retroviruses which have been associated with hematopoietic neoplasms in primates. Two of the viruses, GALV SEATO and GALV-San Francisco (GALV-SF), are associated with myeloid and lymphocytic leukemias, respectively, in apes. Using an assay based on the transient expression of the bacterial gene chloramphenicol acetyltransferase (CAT), we examined the transcriptional activity of GALV-SEATO and GALV-SF. The results suggest that high level expression of GALV is due primarily to cis-acting enhancer sequences. Sequence delineation analysis of GALV-SEATO showed the GALV SEATO enhancer sequences to be located within a 45-bp tandem repeat in GALV SEATO. GALV-SF, which has two- to fivefold lower transcriptional activity, contains only a single copy of the 45-bp element with 6-bp differences from those in the GALV-SEATO enhancer element. This 45-bp element is highly homologous to sequences within the LTRs of several murine leukemia viruses but has not been examined for enhancer function in these retroviruses. Expression of GALV was not restricted to hematopoietic cells but was extraordinarily high in MLA 144 cells, a gibbon ape T-cell line known to be infected with GALV-SF. However, expression of constructs containing the CAT gene directed by GALV-SEATO LTR sequences was similar in uninfected and GALV-infected fibroblasts, indicating the lack of virally encoded or virally induced trans-activating factors capable of increasing expression in these cells. PMID- 3029961 TI - SV40 late promoter: contribution of the initiation site sequences to basal late promoter activity. AB - We have previously shown that the +7 to -53 element of the SV40 late promoter (nt 273 to nt 332) does not have any promoter activity, but is able to stimulate the late promoter activity of the enhancer element. The +7 to -53 element contains several late transcriptional initiation sites and we have shown that its removal results in an increase in the heterogeneity of initiation sites. Furthermore we found that inversion of the +7 to -53 element does not adversely affect the efficiency of late transcription. However, when the +7 to -53 element was inverted, transcription initiated from a single site at nt 302. In fact, we noticed that there is a consensus TATA box signal 26 nt upstream of this single site in the inverted +7 to -53 element. These results may indicate that the ability of +7 to -53 element to function in both orientations is due to the fact that, in both orientations, it possesses sequences capable of fixing the initiation sites of transcription. PMID- 3029960 TI - A restricted component of the Epstein-Barr virus early antigen complex is structurally related to ribonucleotide reductase. AB - The 85-kDa polypeptide previously shown to be associated with the restricted (R) component of the EBV-induced early antigen (EA) complex was subjected to amino acid sequencing analysis. This was accomplished by cyanogen bromide cleavage and by separation of individual peptides by high-pressure liquid chromatography employing reversed-phase C18 column techniques. Two of the isolated peptides, F11 and F13, were subjected to amino acid sequencing and both were found to have significant homology to the postulated protein encoded by the BamHI O right reading frame 2 (BamHI ORF2) of the B95-8 strain Epstein-Barr virus. Computer analysis revealed significant homology between the amino acid sequence of this polypeptide and ribonucleotide reductase, an enzyme previously mapped to this genomic fragment. Amino acid composition analysis also revealed a similar association. These results indicate that the 85-kDa EA(R) polypeptide is associated with a component of the EBV-induced ribonucleotide reductase. PMID- 3029962 TI - Transgenic chickens: insertion of retroviral genes into the chicken germ line. AB - We infected early chicken embryos by injection of wild-type and recombinant avian leukosis viruses into the yolk of unincubated, fertile eggs. The viremic males (designated generation 0 (G-0] were tested for transmission of proviral DNA to their G-1 progeny. Nine of 37 G-0 viremic males were mosiac and proviral DNA was transmitted to their progeny at frequencies varying from 1 to 11%. All of the G-1 progeny examined by restriction enzyme analysis for clonality of proviral junction fragments had one to three simple but different fragments. The proviral DNA was transmitted from G-1 to the G-2 progeny in a Mendelian fashion thus proving that retroviral genes have been inserted into the chicken germ line. One of the viruses is a candidate vector for insertion of foreign genes into the chicken germ line. PMID- 3029963 TI - Molecular cloning and sequence analysis of the human parainfluenza 3 virus gene encoding the matrix protein. AB - The sequence of the matrix (M) protein gene and contiguous intergenic regions of the human parainfluenza 3 virus (PF3) was determined by molecular cloning. The encoded M protein contains 354 amino acids and has a predicted mol wt of 39,506. The M protein amino acid sequence was compared to the homologous proteins from other members of the Paramyxoviridae family. The PF3 protein shared 61% homology with the Sendai virus protein and approximately 35% homology with measles and canine distemper virus proteins. Little homology was observed with respiratory syncytial virus. The M protein appears to be the most highly conserved among the Paramyxoviridae proteins. PMID- 3029964 TI - Gene disruption indicates that the only essential function of the SKI8 chromosomal gene is to protect Saccharomyces cerevisiae from viral cytopathology. AB - We have cloned the SKI8 gene, one of the chromosomal genes that repress replication of M, L-A, and L-BC double-stranded RNA viruslike particles in yeast. The clone was used to map SKI8 to chromosome VII near ade5 and to construct a deletion mutant. The deletion mutant was unable to grow at 8 degrees if and only if M1 double-stranded RNA was present. PMID- 3029965 TI - Sequence analysis of the bovine coronavirus nucleocapsid and matrix protein genes. AB - The 3' end of the 20-kb genome of the Mebus strain of bovine enteric coronavirus (BCV) was copied into cDNA and cloned into the PstI site of the pUC9 vector. Four clones from the 3' end of the genome were sequenced either completely or in part to determine the sequence of the first 2451 bases. Within this sequence were identified, in order, a 3'-noncoding region of 291 bases, the gene for a 448 amino acid nucleocapsid protein (N) having a molecular weight of 49,379, and the gene for a 230-amino acid matrix protein (M) having a molecular weight of 26,376. A third large open reading frame is contained entirely within the N gene sequence but is positioned in a different reading frame; it potentially encodes a polypeptide of 207 amino acids having a molecular weight of 23,057. A higher degree of amino acid sequence homology was found between the M proteins of BCV and MHV (87%) than between the N proteins (70%). For the M proteins of BCV and MHV, notable differences were found at the amino terminus, the most probable site of O-glycosylation, where the sequence is N-Met-Ser-Ser-Val-Thr-Thr for BCV and N Met-Ser-Ser-Thr-Thr for MHV. BCV apparently uses two of its six potential O glycosylation sites. PMID- 3029966 TI - Transfection of human lymphocytes with cloned Epstein-Barr virus (EBV) DNA. AB - Human primary cord-blood lymphocytes were transfected, using the DEAE-dextran technique, with a set of seven largely overlapping clones jointly covering the whole M-ABA Epstein-Barr virus (EBV) genome. Three fragments, cosmids cMB-14 and cM301-99 and plasmid pM966-20, were able to stimulate transient cellular DNA synthesis, blastic transformation, and clumps formation, as well as to prolong the life span from a maximum of 2 weeks in control cultures to up to 6 weeks. The fragments stimulating DNA synthesis also expressed this property in mutual combinations or when combined with cosmid cMSal-A or cM302-21. Their use with any other fragments in cotransfection did not result in further DNA synthesis stimulation. Cosmids cM302-23 and cMSal-B suppressed this effect. Cosmid cM301-99 but not cM302-23 induced transient EBNA-1 formation in about 1% of lymphocytes. Lymphocytes transfected with single fragments or their combinations failed to grow into immortalized cell lines. The results suggest that transient expression of viral functions at levels achievable by transfection is not sufficient for cell immortalization. PMID- 3029967 TI - The single base pair substitution responsible for the Syn phenotype of herpes simplex virus type 1, strain MP. AB - Nucleotide sequences were determined for portions of the genomes of the syncytial (Syn) mutant of herpes simplex virus type 1, strain MP, and the related wild-type strain mP. Comparisons of the nucleotide sequences showed only 1 bp difference between the DNAs of strains MP and mP in the region to which the Syn mutation of MP had previously been mapped. This base pair substitution in MP (at map coordinate 0.737) eliminates a ThaI restriction endonuclease recognition site that is present in mP DNA. Analyses of MP X mP recombinant viruses showed that presence of the ThaI site correlates with the Syn+ phenotype and absence of the ThaI site correlates with the Syn phenotype as predicted. We conclude that the base pair substitution at map coordinate 0.737 is responsible for the Syn phenotype of MP. This mutation could alter translation in four of the six reading frames, causing amino acid substitutions. From only one of these reading frames is a product likely to be expressed. The 338-amino acid polypeptide that could be expressed has features characteristic of membrane-associated proteins, including hydrophobic domains, potential sites for the attachment of N-linked carbohydrate, and a potential cleavable signal sequence. PMID- 3029968 TI - Multiple mutations involved in the phenotype of a temperature-sensitive small plaque mutant of poliovirus. AB - A temperature-sensitive small-plaque mutant of poliovirus type 1, ts247, has been analyzed previously. Several mutations were detected in the P3 region of the genome by analysis of proteins and by T1 oligonucleotide mapping of viral RNA. We have now studied spontaneous reversion of ts247 to the wild-type phenotype. This was found to be a two-step event, reversion to a ts+ phenotype (revertant R247 51) being distinct from acquisition of normal plaque size (revertant R247-12). The mutation responsible for the ts phenotype of ts247, implicated also in virus aggregation and heat lability, could not be detected by biochemical studies. Analysis of homotypic recombinants obtained by crossing ts247 with a guanidine resistant derivative of a temperature-sensitive replicase mutant mapped this mutation to the P1 region or to the 5' end of the P2 region of the genome. The small-plaque phenotype of ts247 and R247-51 was correlated with an abnormality in polypeptide 3C (protease); direct sequencing of viral RNA revealed a U to C change at nucleotide 5658, which altered an isoleucine to threonine in the protease of ts247 and R247-51 but not of R247-12. Two other mutations were present in the region of the genome coding for polypeptide 3D of ts247 and of both classes of revertants. They thus seemed to play no role in the phenotype of ts247. One mutation, an A to G change at nucleotide 7135, was silent at the protein level, whereas the other, an A to G change at nucleotide 6264, determined a major amino acid change from glutamate to glycine in the viral replicase. PMID- 3029969 TI - Guanidine-dependent mutants of poliovirus: identification of three classes with different growth requirements. AB - Four mutants resistant to high (2.0 mM) guanidine were derived from a mutant resistant to intermediate (0.53 mM) levels of this drug. One of these mutants was found to be resistant to high guanidine and was shown to contain a mutation within 2C seen previously in this class of mutants, while lacking the mutation seen in the intermediate parent. The other three mutants were dependent on guanidine for growth and contained the mutation in 2C seen in the parental virus as well as a mutation seen previously in another dependent mutant. Comparison of the newly isolated dependent mutants to two previously described dependent mutants revealed that three classes of dependent mutants which vary in their requirements for optimal growth can be observed. We present a model for the interaction of guanidine with 2C that explains the occurrence of the three classes of dependent mutants. PMID- 3029970 TI - The effect of elevated levels of herpes simplex virus alpha-gene products on the expression of model early and late genes in vivo. AB - The rate of synthesis in vivo and the steady-state level of mRNA of four "model" herpes simplex virus type 1 (HSV-1) genes were measured as a function of high levels of alpha-gene products. The genes studied were ICP4 (alpha), deoxy-UTPase (beta), VP5 (beta gamma), and glycoprotein C (gC, gamma). Accumulation of high levels of alpha proteins was accomplished either by infection with an HSV-1 mutant, temperature-sensitive in ICP4 (ts606) at the nonpermissive temperature then shift-down to permissive temperature, or by infection with wild-type virus under cycloheximide blockage of protein synthesis followed by release. Compared to RNA expression in normal infections, beta gamma and gamma transcription rates were both transiently stimulated under the experimental conditions employed. The greatest effect was seen with the gamma-gC mRNA transcription rates. In addition, at nonpermissive temperatures with ts 606, the amount of expression of gC mRNA was significantly increased over normal early levels, in contrast to the case with the VP5 transcript. The impact of such results on models of HSV gene expression in vivo are discussed. PMID- 3029971 TI - Ca2+ responses in cytomegalovirus-infected fibroblasts of human origin. AB - Cytomegalovirus (CMV) infection of fibroblasts of human origin is associated with a cascade of cellular responses (rounding, "contraction," "relaxation," and enlargement). Since in other systems these cellular responses are regulated by intracellular free Ca2+ activity ([Ca2+]i), we measured intracellular Ca2+ responses to CMV infection. At relatively high multiplicities of infection (m.o.i), an influx of Ca2+ was observed within the first hour after CMV infection (p.i.) at which time it was at its maximum rate. Both the time of occurrence and the magnitude of this Ca2+ influx were dependent on the calculated input m.o.i. In CMV-infected cells, [Ca2+]i rose gradually from 80 nM at 0 hr to 174 nM at 48 hr p.i. (about 2.7 times the [Ca2+]i found in mock-infected cells at this time). At 8 and 12 hr p.i. CMV-infected cells consistently contained a somewhat greater level of 45Ca2+ than mock-infected cells, despite the fact that there was only a small increase in [Ca2+]i between CMV and mock-infected cells in the same period. This observation suggests that there may be significant amounts of Ca2+ taken up into intracellular stores. This Ca2+ in intracellular stores may, at later times after infection, contribute to the increase in [Ca2+]i observed from 12 to 48 hr p.i. Ca2+ influx blockers, such as nifedipine and verapamil, inhibited the rise in [Ca2+]i. The increase in [Ca2+]i in response to CMV infection was shown to be dependent on the m.o.i., require infectious virus, and occur under conditions consistent with the expression of immediate-early CMV genes. The capability of inducing such Ca2+ responses was conserved among three laboratory strains of CMV. The CMV-induced Ca2+ responses may be related not only to the development of CMV cytopathology, but also to the replication of CMV, since in other studies cyclic nucleotide modulators and Ca2+ influx blockers were found to inhibit the replication of CMV. PMID- 3029972 TI - Replicative events in hepatitis A virus-infected MRC-5 cells. AB - Replication of hepatitis A virus (HAV) in MRC-5 cells was studied under one-step growth conditions. Viral replication neither interfered detectably with cellular DNA, RNA, and protein synthesis, nor could cytopathologic changes be recorded over a prolonged period of incubation. Synthesis of mature, infectious HAV particles could be detected as early as 2-4 days p.i. and occurred at a maximal rate around 8 days p.i., shortly before infectivity titers reached a plateau. Synthesis of total viral RNA, of positive-strand genomic RNA, of viral mRNA, as well as of negative-strand RNA followed the same pattern. By Day 14 p.i., when active HAV replication had developed into persistent infection, synthesis of viral RNA declined to background levels. The mechanism(s) guiding active HAV replication into a state of persistent infection could not be positively defined. Yet there exists the possibility that this is brought about by down regulation of viral RNA synthesis. Whether this is related to the appearance of a subgenomic viral RNA molecule about 2000 nucleotides in length and detected in association with ribosomes on Days 7 and 10 p.i. remains to be shown. PMID- 3029973 TI - Sequences of Chandipura virus N and NS genes: evidence for high mutability of the NS gene within vesiculoviruses. AB - The nucleotide sequence of the 3' end of the genome of Chandipura (CHP) virus, including the complete sequences of the nucleocapsid (N) and phosphoprotein (NS) genes was determined, principally from cloned cDNAs of the N and NS mRNAs. The NS mRNA of CHP virus is 908 bases in length and encodes a protein of 293 amino acids. Comparison of the CHP virus NS protein sequence with those of vesicular stomatitis virus of the New Jersey serotype (VSV (NJ)) and of the Indiana serotype (VSV (IND] revealed homologies of only 23 and 21%, respectively, with no consecutive stretches of more than four amino acids identical among the three sequences. As with the two VSV serotypes, the highest homology between the NS proteins of CHP and VSV was in a 20-amino acid region near the carboxy termini of the proteins. Of the potential phosphorylation sites, there are eight conserved serine or threonine residues among the three sequences. Despite the dissimilarity among primary sequences of the NS proteins, their overall structure, as assessed by amino acid composition and by the relative hydropathicities of the sequences, has been conserved throughout evolution. The N mRNA of CHP virus is 1291 bases long and encodes a protein of 422 amino acids. In contrast to the NS protein, the CHP N protein is at least 50% homologous to the N proteins of each of the VSV serotypes. We have identified a region near the center of these N protein sequences which is conserved among members of both the rhabdovirus and paramyxovirus families. This extent of conservation of the N protein sequences underscores the high rate of mutability of the NS protein sequences among the vesiculoviruses. PMID- 3029974 TI - DNA sequences which regulate the expression of the pseudorabies virus major immediate early gene. AB - It has been shown previously that the transcription of herpes simplex virus (HSV) immediate early (IE) genes is transactivated by a component of the virus particle. The trans-inducing factor (TIF) is known to be polypeptide Vmw65. Infection with pseudorabies virus (PRV), a related herpesvirus, does not increase expression from HSV IE regulatory sequences (W. Batterson and B. Roizman, 1983, J. Virol. 46, 371-377). To examine the control of the PRV IE gene and possible sequence specificity of a TIF, the 5' terminus of the PRV major IE transcript was mapped and hybrid plasmids containing PRV upstream sequences linked to the HSV-1 TK gene were constructed. Gene expression under the control of PRV IE or HSV-1 IE gene 3 upstream regions were compared using transient expression assays. It was found that infection with uv-irradiated PRV did not stimulate expression from PRV IE or from HSV-1 IE gene 3 upstream regions, indicating that PRV did not possess an effective TIF. Infection with uv-treated HSV-1, or cotransfection with a plasmid which encodes Vmw65, stimulated expression from both PRV and HSV IE gene upstream regions. The nucleotide sequence of the 5' end of the PRV transcript and its upstream region was determined. This region was, in overall structure, unlike the upstream regions of HSV IE genes but showed a strong similarity to the enhancers of human and murine cytomegaloviruses (HCMV and MCMV). In particular, a reiterated 15-bp element of the PRV upstream region was homologous to a conserved, repeated sequence element found in both HCMV and MCMV enhancer regions and was also related to the "TAATGARATTC" motif found upstream of all HSV IE genes. Thus a conserved sequence element occurs upstream of IE genes in four herpesviruses with different genome structures and diverse biological properties. PMID- 3029975 TI - Characterization of Kunjin virus RNA-dependent RNA polymerase: reinitiation of synthesis in vitro. AB - RNA-dependent RNA polymerase (RDRP) activity was characterized in a cytoplasmic extract of Kunjin virus-infected Vero cells at 24 hr. The activity was influenced, possibly indirectly, by the length of prior treatment of infected cells with actinomycin D; however, 6 micrograms/ml actinomycin D and 10(-5) M alpha-amanitin in the RDRP assay had no effect. The replication complex was membrane-bound and Mg2+ was essential for RDRP activity. Incorporation was more dependent on exogenous UTP and GTP than ATP or CTP. The specific activity was low, and rate of incorporation of GMP decreased as the period of assay was increased; however, incorporation of label lasted for at least 60 min. RNA products were fractionated by LiCl precipitation, and kinetic studies showed that the sequence of accumulation of label was the same as that observed in vivo, viz., RI----RF----44 S RNA; limited reinitiation was also observed. This sequence of labeling also indicated that the in vitro RDRP activity was due to an enzyme capable of elongation, release, and reinitiation of Kunjin RNA synthesis and not merely end labeling or elongating preexisting RNA molecules. No labeled bands in urea-polyacrylamide gels were observed using extracts from mock-infected cells and hence the three RNA products of assays were readily identified in a single gel. The replication complex was still active after treatment with nonionic detergent, but no labeled 44 S RNA was detected in gels, even in the presence of RNasin in the assay which inhibited some nuclease activity. Antibodies to flavivirus-specific nonstructural proteins were preincubated with infected cell extracts in the presence and absence of detergent but no inhibition of RDRP activity was observed. However, anti-dsRNA plus detergent blocked activity by as much as 78% and label was found only in RF. PMID- 3029976 TI - Restriction enzyme map of herpesvirus of turkey DNA and its collinear relationship with Marek's disease virus DNA. AB - The genome of herpesvirus of turkey (HVT) was shown to consist of long and short unique regions flanked by inverted repeats (J. Cebrian, Kaschka-Dietrich, C., Berthelot, N., and Sheldrick, P., 1982, Proc. Natl. Acad. Sci. USA 79, 555-558). In this paper we report the construction of the linkage map of HVT DNA for BamHI, HindIII, and PstI restriction endonucleases. The maps were constructed by hybridization of 19 cloned BamHI fragments of HVT DNA to electrophoretically separated digests of genomic DNA. Our results indicate that the terminal and internal inverted repeats (TRL and IRL) flanking the long unique sequences (UL) are spanned by BamHI-F fragment and a -F-related terminal fragment, respectively, whereas the terminal and internal inverted repeats (TRS and IRS) flanking the short unique sequences (US) are mostly contained in BamHI-A fragment. Both BamHI A and -F showed a heterogeneity in size, suggesting the presence of amplification of certain sequences in the inverted repeats. We also report that the HVT genome is collinear with the genetically related Marek's disease virus (MDV) genome, as determined by hybridization of labeled cloned HVT DNA fragments with electrophoretically separated MDV DNA fragments. PMID- 3029977 TI - Biochemical characterization of an aphthovirus type C3 strain Resende attenuated for cattle by serial passages in chicken embryos. AB - We have compared several aspects of an aphthovirus strain attenuated for cattle (C3R-O/E) with the original strain (C3Res) from which it was derived after serial passages in chicken embryos. Biochemical differences detected by protein analysis in regular polyacrylamide gels (SDS-PAGE) and on electrofocusing gels (NEPHGE) suggest the presence of mutations throughout the genome. Changes were located in coat proteins VP1 and VP3 and in the polymerase precursor P100 (P3/ABCD). No other differences were found at the protein level by means of the techniques used. Polypeptide P100 of the attenuated strain showed a faster electrophoretic mobility in SDS-PAGE with respect to that of the wild-type strain, and the change seems to be located on its amino terminus half. Several functional differences were also found between the two viruses. Both strains grew equally well in BHK cells reaching roughly similar titers in plaque assays. However, the wild-type strain maintained its titer in cells of bovine origin (BK), whereas the titer of C3R-O/E strain decreased approximately one log in this cell system; moreover, plaques elicited by the attenuated strain were much smaller than the ones produced by C3Res. A diminution in the rate of RNA synthesis induced by C3R-O/E in BK cells compared with that of the wild-type strain was also detected; this trait was not observed in BHK cells. A delay in the kinetics of RNA synthesis was also detected in this strain. The virus yield of attenuated strain in BK cells was four times lower than in BHK cells. PMID- 3029978 TI - Purification and properties of virus particles, infectious subviral particles, and cores of bluetongue virus serotypes 1 and 4. AB - Effective purification methods have been developed for virus particles, infectious subviral particles (ISVP), and virus cores of bluetongue virus (BTV) serotypes 1 and 4. The purified particles were analysed by indirect ELISA or PAGE using either silver staining, or fluorography of [35S]methionine-labelled preparations. No significant contamination with host cell proteins, or with the majority of BTV nonstructural proteins was detectable in any of the particle preparations. In addition to the two major outer capsid and five core proteins previously described, the purified virus particles of both serotypes were consistently found to contain small amounts of BTV protein NS2, previously regarded as exclusively nonstructural. This protein could be removed from the particle surface by treatment with a combination of chymotrypsin and sodium N lauroyl sarcosinate, which also resulted in the cleavage of the larger of the two major outer capsid components (protein VP2). Two of the cleavage products of VP2 and the whole of the other major outer capsid component (protein VP5) formed a modified outer capsid layer in the resultant ISVP. These subviral particles were as or more infectious than the intact virus particles but had lost haemagglutinating activity. The core-associated RNA polymerase remained inactive in ISVP. PMID- 3029980 TI - Large-scale rearrangement of homologous regions in the genomes of HCMV and EBV. AB - The 20,349-bp sequence of the human cytomegalovirus (HCMV) HindIII F fragment has revealed eight open reading frames with homology to herpes simplex virus (HSV) and/or Epstein-Barr virus (EBV). With respect to EBV, these homologous genes can be divided into two blocks: one block contains three genes, including the DNA polymerase and glycoprotein B, and the other block contains five genes of unknown function. Although the relative organisation of genes within each block is identical in HCMV and EBV, the relative position of each block within the two genomes differs: in HCMV the two blocks are present directly adjacent to each other, whereas in EBV they are found 92 kb apart. This suggests that a genetic rearrangement has occurred in this region. Transcription analysis of the glycoprotein B gene is presented and the evolutionary relationship between the genomes of HCMV, EBV, and HSV is discussed. PMID- 3029979 TI - The effects of leukemosuppressive immunotherapy on bone marrow infectious cell centers in AKR mice. AB - The bone marrow of AKR mice is the richest source of infectious ecotropic cell centers (ICCs) during the neonatal period. The bone marrow ICCs reside in a low density population expressing high levels of viral glycoprotein (gp71) and Class I histocompatibility antigens (H-2Kk). In addition, ICCs are enriched in the lymphoid band of Ficoll-Hypaque-fractionated bone marrow, the adherent population of nylon wool separated cells and among the low-density subpopulation of Percoll fractionated marrow. The observed dichotomy between viral antigen expression and actual virus production suggests that actively cycling cells may be the primary virus producers in the AKR bone marrow. The phenotypic and physical data indicate that bone marrow stem cells and/or prothymocytes may be among the initial virus producing cells in the AKR bone marrow. Leukemosuppressive antiviral immunotherapy delays the appearance of ICCs in the bone marrow but does not exert any major long-term changes on the populations of cells present. PMID- 3029981 TI - Synergistic role of staphylococcal proteases in the induction of influenza virus pathogenicity. AB - Several strains of Staphylococcus aureus have been found to secrete proteases that activate infectivity of influenza virus by proteolytic cleavage of the hemagglutinin. The enzymes of the bacterial strains Wood 46 and M 86/86 have been characterized in some detail and were found to be serine proteases. In their substrate specificities and inhibitor sensitivities they proved to be similar to, but not identical with trypsin and plasmin. The hemagglutinin of an individual virus strain could be cleaved by the proteases of some but not all staphylococcal strains, and a given enzyme could cleave only some but not all hemagglutinins analyzed. When mice were coinfected intranasally with the appropriate strains of influenza virus and S. aureus, the hemagglutinin was readily activated allowing multiple cycles of virus replication in the lung. Under these conditions, the animals came down with a fatal disease exhibiting extended lesions in the lung tissue. In contrast, after infection with virus or bacteria alone, there were no significant pathological changes. When the staphylococcal strain did not contain a protease that was able to activate the hemagglutinin of the coinfecting virus strain, the animals did not exhibit disease. These observations demonstrate that coinfecting bacteria can play an essential role in the development of influenza pneumonia by providing a protease suitable for cleavage activation of the hemagglutinin. PMID- 3029982 TI - The carboxyl 35 amino acids of SV40 Vp3 are essential for its nuclear accumulation. AB - To identify the moiety responsible for nuclear localization of the SV40 structural protein Vp3 in its natural environment, a transfection vector containing the entire coding regions of Vp2, Vp3, and agnoprotein, and one-third of the coding region of Vp1, was constructed. Several mutations were introduced into the plasmid and the subcellular distribution of Vp3 or mutant Vp3 was examined following DEAE-dextran-mediated DNA transfection into TC7 cells. Our study shows that Vp3 is synthesized and is transported into the nucleus in the absence of Vp2, agnoprotein, and intact Vp1. However, in the absence of its carboxyl-terminal 35 amino acids, the truncated Vp3 is limited to a cytoplasmic and perinuclear accumulation. Thus, the carboxyl 35 amino acids of Vp3 are required for its nuclear localization and may contain a nuclear accumulation signal. PMID- 3029983 TI - Characterization of the Epstein-Barr virus-induced early polypeptide complex p50/58 EA-D using rabbit antisera, a monoclonal antibody, and human antibodies. AB - A polypeptide complex (p52) belonging to the D-subspecificity of the EBV-induced early antigens (EA-D) was purified from chemically induced P3HR-1 cells. Rabbit antisera raised against the isolated polypeptides reacted with components of EA-D as could be shown by indirect immunofluorescence and immunoperoxidase staining of IdU-induced EA positive Raji cells, ELISA, and immunoblotting. In one-dimensional immunoblots the rabbit antisera detected a predominant polypeptide complex of 52 kDa. Two-dimensional immunoblots prepared with proteins from IdU-induced Raji cells showed that the rabbit sera detect three series of polypeptides of 52 kDa (pl 8.5-6.2), 55-58 kDa (pl 6.2-4.5), and 48-50 kDa (pl 6.0-4.5). These three groups of polypeptides could also be identified by 50 high titered anti-EA-D positive human sera and a specific monoclonal antibody (R3) as being the main components of EA-D in Raji and B95-8 cells. All polypeptides of the p50/58 complex showed DNA binding properties either by themselves or by an interaction with other proteins. When TPA or IdU-induced Raji cells were labeled with 2Pi, two phosphorylated polypeptides pp50 and pp58 could be immunoprecipitated with the rabbit sera and a high anti-EA titered human serum. The time course of the synthesis of polypeptides associated with the EA-D complex was studied by 2-D immunoblots: EA polypeptides of 52 kDa appeared as early as 6 hr after the addition of IdU to Raji cells in culture, polypeptides of 55-58 and 48-50 kDa after 18 and 25 hr, respectively. The coordinated appearance of these groups of polypeptides and their similar size and reactivity with human sera and rabbit antisera produced against the isolated p52 as well as with a monoclonal antibody (R3) suggested that most of these polypeptides are derived from post translational modifications of one or a few initially synthesized polypeptides, possibly p52. Phosphorylation seems to be at least one possibility of post translational modification. PMID- 3029984 TI - Production and characterization of the neutralization antigen VP2 of bluetongue virus serotype 10 using a baculovirus expression vector. AB - DNA representing RNA segment 2 of bluetongue virus (BTV) serotype 10, corresponding to the gene that codes for the BTV neutralization antigen VP2, has been inserted into a baculovirus transfer vector in lieu of the 5' coding region of the polyhedrin gene of Autographa californica nuclear polyhedrosis virus (AcNPV). After cotransfection of Spodoptera frugiperda cells with wild-type AcNPV DNA in the presence of the derived recombinant transfer vector DNA, polyhedrin negative recombinant baculoviruses were recovered. When S. frugiperda cells were infected with one of these recombinant viruses, a protein that was similar in size and antigenic properties to the BTV VP2 protein was synthesized. Antibodies raised in mice or rabbits to the baculovirus expressed VP2 protein neutralized the infectivity of BTV-10 virus and to lesser extents BTV serotype 11 and 17 viruses but not BTV-13 virus. PMID- 3029985 TI - In vitro synthesis of two polypeptides from a nonstructural gene of coronavirus mouse hepatitis virus strain A59. AB - The complete nucleotide sequence of nonstructural gene 5 of coronavirus mouse hepatitis virus (MHV) strain A59 has been determined. This sequence contains two potential open reading frames which overlap by five nucleotides. The putative protein products predicted from the sequence are a basic 13,000-Da polypeptide and a 9600-Da polypeptide containing an unusually long hydrophobic amino terminus. RNAs transcribed in vitro from DNAs containing each of the open reading frames in pGEM vectors direct the synthesis in vitro of polypeptides of the sizes predicted by the sequence. An RNA transcript containing both of the open reading frames directs the synthesis primarily of the polypeptide corresponding to the downstream open reading frame. These data suggest that MHV-A59 messenger RNA 5 potentially encodes two proteins and may be preferentially translated from an internal AUG initiation codon. PMID- 3029986 TI - Identification and sequence of the gene encoding gpIII, a major glycoprotein of varicella-zoster virus. AB - The genome of varicella-zoster virus (VZV) encodes three major glycoproteins, two (gpI and gpII) having been mapped and sequenced, which carry epitopes capable of eliciting neutralizing antibodies. The product of the third major glycoprotein gene (gpIII) was purified, and seven consecutive amino acids at its N-terminus were identified. A degenerate pool of oligonucleotides based upon this sequence was used as a probe to localize the gpIII gene to the HindIII B fragment of the VZV genome. An analysis of the DNA sequence from this region revealed an open reading frame (ORF) encoding 841 amino acids. Rabbit antisera against three synthetic peptides derived from the putative gpIII gene recognized a protein which comigrated with gpIII in Western blots and immunoprecipitation analysis. Preclearing with a monoclonal antibody to gpIII specifically abolished immunoprecipitation of this protein. Also a polypeptide translated from mRNA selected by the putative gpIII gene could be immunoprecipitated by the anti peptide sera. Therefore, we conclude that gpIII is encoded by the identified ORF in HindIII B. In addition, gpIII is implicated as essential for the cell-to-cell spread of VZV. PMID- 3029987 TI - Nucleotide conservation of endogenous ecotropic murine leukemia proviruses in inbred mice: implications for viral origin and dispersal. AB - Nucleotide sequence analysis of the ecotropic murine leukemia proviruses of AKR, BALB/c, and C57BL/6 mice indicated that these viral genomes differ from each other in less than 0.5% of their sequenced nucleotides, whereas they differ from the laboratory Moloney, Friend, or RadLV viruses or a partial ecotropic provirus found in wild mice by 8-22% of their sequenced nucleotides. The limited variation of endogenous ecotropic proviruses found in these common mouse strains indicates that few cycles of virus replication separated the introduction of the ecotropic endogenous retroviruses into the germlines of the progenitors of these now divergent mouse strains, and is consistent with the hypothesis that these common inbred strains were derived from a pool of very few mice, at least one of which was infected with an ecotropic murine leukemia virus. Ecotropic germline proviruses now found in common inbred mice most likely derive from germline reintegrations of the viral progeny of that initial single infection. PMID- 3029988 TI - Identification of an Epstein-Barr virus glycoprotein which is antigenically homologous to the varicella-zoster virus glycoprotein II and the herpes simplex virus glycoprotein B. AB - The Epstein-Barr virus (EBV) antigenic homologue of the varicella-zoster virus glycoprotein II and the herpes simplex virus (HSV) glycoprotein B (gB) was identified through cross-reactivity with anti-glycoprotein II and anti glycoprotein B peptide sera. The homologue is the previously characterized EBV glycoprotein, with an apparent molecular weight of 125,000 Da, which is synthesized late during productive EBV infection and appears to be encoded by the BamHI A EBV fragment. This glycoprotein, but not other EBV proteins, reacted with the antisera in immunoprecipitation experiments and by ELISA. In addition, absorption of the sera with the purified EBV 125-kDa glycoprotein removed the cross-reacting antibody. Whether the EBV gB homologue has the same biological functions associated with HSV gB has yet to be determined. PMID- 3029989 TI - Expression of the poliovirus genome from infectious cDNA is dependent upon arrangements of eukaryotic and prokaryotic sequences in recombinant plasmids. AB - The introduction of a cDNA copy of poliovirus type 1 (Mahoney) into cultured primate cells results in the production of infectious virus. The level of infectious virus can be increased by incorporation of eukaryotic signals of transcription and replication. We have utilized the SV40 DNA sequence coding for the early and late promoters, the SV40 origin of replication, and the enhancer elements, along with the cDNA of poliovirus, to determine the important parameters for the level of infectious virus produced following transfection. Although plasmid replication increases the level of infectivity, the major determinant of infectivity is promoter activity. PMID- 3029990 TI - Identification of putative polymerase gene product in cells infected with murine coronavirus A59. AB - The virion RNA of mouse hepatitis virus, strain A59 (MHV-A59) is believed to be the mRNA for the viral RNA-dependent RNA polymerase. The cell-free translation of virion RNA results in the synthesis of two predominant products p220 and p28 (M. R. Denison and S. Perlman, 1986, J. Virol. 60, 12-18). p28 is a basic protein and is readily detected by two-dimensional gel electrophoresis. When infected cells and isolated virions were assayed for this protein by two-dimensional gel electrophoresis, p28 could be detected in infected cells labeled at late times after infection, but not at early times or in purified virions. p28 represents the first protein product of the putative coronavirus polymerase gene to be identified in infected cells. PMID- 3029991 TI - [Paraneoplastic Cushing's syndrome in metastasizing medullary cancer of the thyroid gland]. PMID- 3029992 TI - [Inhibitors of angiotensin convertase (inhibitory ACE) in chronic heart failure]. PMID- 3029993 TI - [Incidence of metastatic lesion of different groups of the intrathoracic lymph nodes in lung cancer (based on research data on surgical preparations)]. AB - The study was concerned with the frequency of metastatic spread of lung cancer to intrathoracic lymph nodes versus lung lobe distribution of malignancy in 299 cases of surgical treatment. In cases of primary tumor in any lobe of both lungs, metastases were detected primarily in the pulmonary and bronchopulmonary lymph nodes. A decreasing frequency of metastases was observed from the intrapulmonary lymph nodes to those of the root of the lung and mediastinum. PMID- 3029995 TI - [Serous meningitis in Odessa in the summer-fall period of 1984]. PMID- 3029994 TI - [A new kind of dietetic bread enriched with dietary fiber]. AB - A new sort of dietetic bread has been developed with a 4-fold higher content of food cellulose fibers and a 25% lower fuel value. Such effect was achieved due to the use of qualitatively different sources of food fibers--wheat bran and methyl cellulose (MLI-100). The chemical composition of the new sort of bread has been presented and certain categories of patients have been specified who can use this bread in their diet. PMID- 3029996 TI - [Extra-adrenal action of corticotropin (a review of the literature)]. PMID- 3029997 TI - [Neutrophil peroxidase activity in the peripheral blood of patients with rubromycosis of the feet]. PMID- 3029998 TI - [Rotaviruses: their structure, chemical composition and biological properties]. PMID- 3029999 TI - [Use of natural sorbents for isolating enteroviruses and hepatitis A virus from water samples]. AB - The data of the study on optimal conditions for the use of a mineral sorbent, bentonite, for isolation of hepatitis A virus and enteroviruses from water bodies are presented. PMID- 3030000 TI - [Increase in the specificity of immunoenzyme analysis by using a monoclonal antibody-based conjugate]. AB - Examinations by EIA of laboratory cultures of various orthopoxviruses and specimens from human-monkey pox cases demonstrated the advantages of a monoclonal preparation over the peroxidase polyclonal conjugate prepared from hyperimmune vaccinia antiserum. PMID- 3030001 TI - [Etiological significance of rotaviruses in acute intestinal diseases in children]. AB - The paper presents the results of virological, serological, and epidemiological examinations of patients with acute enteric infections of obscure etiology carried out for detection of rotavirus gastroenteritides. Rotaviruses have been found to play a certain role in generating acute gastroenteritides in infants under 3 years, mostly among infants, of 1-2 years, the least among those of 0-6 months of age. Adult family members were the source of infection for the children. The season of infection falls on October-December. PMID- 3030002 TI - [Morphology of coronaviruses from different species of monkeys in the Sukhumi nursery]. AB - The ultrastructure of coronaviruses from 46 out of 111 monkeys examined (baboons, macaques, green monkeys, langurs) was studied by negative staining in homogenates of different parts of the intestinal tract, pancreatic gland, liver, kidneys, lungs, heart, and brain. Because of marked pleomorphism of coronaviruses it is suggested that morphological variants of the viruses may be distinguished. No relationship between pleomorphism of virus particles and species differences of monkeys and their organ pathology was established. Morphological signs distinguishing simian coronaviruses from those of man were noted. PMID- 3030004 TI - [Enhancement of alphavirus reproduction in cell cultures at an alkaline pH]. AB - Replication of different alphaviruses, among them Semliki Forest, Sindbis, and Venezuelan equine encephalomyelitis viruses, was studied in chick embryo fibroblasts and hamster BHK cells at different pH in a range of 6.8 to 7.9. Incubation of the infected cell cultures in a mild alkaline medium (pH 7.5-7.9) resulted in a marked activation of both one-cycle and multi-cycle reproduction of the alphaviruses under study as compared with that in the neutral medium (pH 6.8 7.2). At an alkaline pH, the rate of amplification and level of synthesis of viral proteins in the infected cells increased markedly, while virus adsorption on cell receptors decreased by 30-50%. It is recommended to use nutrient media with mild alkaline pH for stabilization of alphaviruses reproduction in cell cultures. PMID- 3030003 TI - [Development of an immunoenzyme test for determining herpes simplex type 1 virus using monoclonal antibodies]. AB - Conditions of the enzyme immunoassay for the detection of HSV-I by the "sandwich" method at all stages of the testing were optimized using monoclonal antibodies (monoAB). When identical concentrations of monoclonal antibodies and durations of their adsorption on polystyrene plates were used, the signal in EIA (A405) with carbonate buffer was by 30% higher than that for PBS. A similar increase in the signal was observed at a temperature of incubation of 37 degrees C as compared with that at 4 degrees C. Even during 18-hour incubation at 37 degrees C no inactivation of monoAB occurred. It was further shown that saturation of the surface of wells with protein was achieved at a concentration of monoAB 5-10 micrograms/ml, and under these conditions the optimal dilution of the conjugate was that corresponding to antibody concentration of 1.5 micrograms/ml. When ABDC peroxidase and 5-aminosalycilic acid were used as the substrate, the presence of HSV-I was regularly detected even with a concentrated virus preparation diluted 500-1000-fold (corresponding to approximately 5.0 1g TCID50). PMID- 3030005 TI - [Immunoenzyme analysis in the diagnosis of human rotavirus gastroenteritis (methodological aspects)]. PMID- 3030006 TI - [Potentials of the passive hemagglutination reaction as a method for the rapid detection of orthopoxviruses and their antibodies]. PMID- 3030007 TI - Side effect of captopril and enalapril. PMID- 3030009 TI - Human immunodeficiency virus and hepatitis testing of old bottles of plasma. PMID- 3030008 TI - Dementia and ataxia in a patient with AIDS. PMID- 3030010 TI - AIDS vaccine trial volunteers can be found. PMID- 3030011 TI - [Fibronectin in the sputum in malignant and inflammatory diseases of the respiratory tract]. AB - Fibronectin , a glycoprotein, which is detectable in the pericellular space, can not be found on the surface of malignant transformed cells, but in the surrounding medium in a higher concentration. Which this finding, it was possible to differentiate between inflammatory and malignant genesis of pleural fluid and ascites. In the sputum and in the bronchial lavage different concentrations can be found too: In carcinomas the concentration on the average is much higher but not constant. This glycoprotein can therefore be used as a further tool in the search for neoplasms in the bronchial tree. PMID- 3030012 TI - Interaction of miconazole, ketoconazole and itraconazole with rat-liver microsomes. AB - The interaction of the antimycotics miconazole, ketoconazole and itraconazole with liver microsomes from untreated rats or from rats pretreated with phenobarbital or 3-methylcholanthrene, gave rise to type II difference spectra. The interactions of the antimycotics with control, phenobarbital-induced or 3 methylcholanthrene-induced microsomes were biphasic, except for the monophasic binding of ketoconazole to phenobarbital-induced microsomes. The N-demethylation of N,N-dimethylaniline, the O-demethylation of p-nitroanisole and the hydroxylation of aniline in microsomes from untreated and inducer-treated rats were lowered by miconazole and ketoconazole, the former being the more potent inhibitor. Control microsomes were less sensitive than induced microsomes. Itraconazole was almost devoid of inhibitory properties. The three antimycotics were non-competitive (mixed) inhibitors of enzyme activities in phenobarbital induced microsomes. The Ki values were of the same order of magnitude as the Ks values, except for itraconazole. For the latter drug, Ki values were much greater than could be expected from the spectral studies. It is concluded that the antimycotics affect microsomal enzyme activities via a direct interaction of an azole-nitrogen with the haem group of cytochrome P-450. The interaction with mammalian cytochrome P-450 decreases from miconazole greater than ketoconazole much greater than itraconazole and is much weaker than the interaction of the antimycotics with yeast cytochrome P-450. PMID- 3030013 TI - [Mechanisms of intestinal absorption of nutrients]. AB - The nutrient uptake from the intestinal lumen into the distributing blood circulation is mediated by the epithelial cell of the small intestine. The transfer process through this distinctly polar cell consists of three partial events: entrance of substances through the brush-border membrane, traversal of a metabolic active intracellular space and exit through the baso-lateral membrane. The fundamental transfer mechanisms--simple diffusion, facilitated diffusion, antiport and symport systems, electroneutral and electrogenic processes--are described. The significance of nutrient metabolization for transport processes is discussed: proton secretion by the epithelial cell coupled to the glucose and lactate metabolization is quoted as an illustration. The "acid microclimate" resulting from this proton secretion on the mucosal surface has a significant influence on weak-electrolyte absorption. This effect was clearly demonstrated for in vitro uptake of nicotinic acid into the intestinal tissue. It can be assumed that--similar to the role of a Na+-gradient--the proton gradient on the surface of absorptive epithelia is highly significant as a driving force of nutrient absorption. PMID- 3030014 TI - [Effects of the autonomic nervous system in supraventricular arrhythmia]. AB - Influences of the autonomic nervous system may cause modifications of the initiation, continuation and discontinuation of supraventricular rhythm disturbances. Whether or not clinically relevant bradycardia syndromes (such as the hypersensitive carotid sinus syndrome and the sick sinus syndrome) also may exist primarily on a functional basis, remains to be proven. Organic pathology, whether it is clinically apparent or not, probably provides the basis for the activation of influences on the autonomic nervous system. In atrial fibrillation and atrial flutter the influence of the autonomic nervous system is apparent in a form of the vagotonically and sympathotonically evoked paroxysms. In the case of persistent atrial fibrillation vagal and sympathotonically transmitted changes in the AV-dromotropy determine the mode of heart rate. Junctional re-entry tachycardias and re-entry tachycardias under the influence of accessory pathways, are potentially triggered by mechanisms, as transmitted by the autonomic nervous system. Furthermore, such autonomic influences are responsible for changes in the conducting velocity and the refractory properties, as produced by parallel conducting pathways. Hemodynamic changes themselves, which are induced at the very beginning of the tachycardia, may activate other electrophysiological mechanisms and thereby cause a modification in the type of tachycardia in question. PMID- 3030015 TI - [The sympatho-adrenal system. Implications for the pathogenesis, diagnosis and therapy of cardiac arrhythmias]. AB - Changes in biosynthesis, storage, release and inactivation of the catecholamines noradrenaline and adrenaline, as well as in the dynamic regulation and localization (internalization/externalization) of post- and presynaptic alpha- and beta-receptors and their subpopulations have implications for the pathogenesis of cardiac arrhythmias. Circulating levels of plasma catecholamines provide a reliable biochemical index for sympatho-adrenal activity and reactivity as well as for diagnosis and prognosis of these arrhythmias. Therapeutic interventions should consequently prefer blockade of myocardial alpha- and/or beta-adrenoceptors, especially after postischaemic arrhythmias with externalization of beta-adrenoceptors. PMID- 3030016 TI - [Cooperativity of viral oncogenes in the induction of leukemia in an animal model]. PMID- 3030017 TI - [Cell receptors and tumor markers]. PMID- 3030018 TI - [Special tumors of childhood]. PMID- 3030019 TI - [Receptor mediated modification of cell growth in vitro--kinetics and description of the morphological receptor]. PMID- 3030021 TI - [Cell receptors--functions and deficiencies]. PMID- 3030020 TI - [Receptor, marker and cell membrane: what value has additional morphological and molecular biology analysis of the cell membrane?]. PMID- 3030022 TI - Immunocytochemical demonstration of neuron-specific enolase for differential diagnosis of retroperitoneal tumours in children. AB - Six patients with malignant retroperitoneal tumours causing considerable diagnostic difficulties are reported. In a retrospective investigation the presence of neuron-specific enolase (NSE) was analysed immunocytochemically in specimens of these tumours. In all the patients who had finally got the definite diagnosis neuroblastoma, tumour specimens had a strong reaction for NSE whereas all other tumours did not react. An analysis of NSE already at the primary investigation could have led to an earlier definite diagnosis. PMID- 3030023 TI - Vena occlusive disease simulated by a metastasis of gastric cancer during pregnancy. AB - A 20-year-old woman was treated because of her complaint and symptoms which were thought to be the results or complication of her pregnancy. Based on the cardinal symptom inferior cavography was carried out, and the diagnosis of occlusion of inferior vena cava was established. The woman was treated accordingly but died after a short period. The patient's family supposed neglection and therapeutic misadventure and raised an objection to the medical treatment. Forensic autopsy was carried out to clear up the case. Autopsy showed her basic disease to be gastric cancer with several metastases in the liver. One of them was in the lobus caudatus Spigeli and compressed the inferior vena cava. The compression caused a turbulence in the given contrast material so that thrombosis was suggested. The diagnosis was based on this misleading sign. Negligence was not verified. PMID- 3030024 TI - [Hydroxylapatite ceramic (HA-ceramic) in autogenous tooth transplantation. An intermediate report]. PMID- 3030025 TI - [Hydroxylapatite ceramic (HA-ceramic) in preprosthetic surgical therapy. Procedures for severe mandibular atrophy]. PMID- 3030026 TI - [Infectious bovine rhinotracheitis. Kinetics of antibody in a dairy herd]. PMID- 3030027 TI - Vaccination against Aujeszky's disease by different routes using live attenuated and inactivated virus vaccines in pigs with or without passive immunity. PMID- 3030029 TI - [Toxigenic phages of Corynebacterium diphtheriae]. PMID- 3030028 TI - [Pharmacological analysis of the ontogenetic traits of the functional receptor activity of glial and neuronal GABA uptake and GABA-T activity in the rat hippocampus]. AB - Studies have been made on the effect of GABA mimetics on epileptogenic focus in the hippocamp of growing rats. By the pattern of inhibition of this focus by these drugs, it is possible to evaluate the degree of functional maturation of the activity of GABA receptors, glial and neuronal uptake of GABA, and the activity of GABA-T. Higher sensitivity of GABA receptors in younger animals was found together with a higher functional role of glial uptake in GABA utilization. Functional activity of GABA-T and neuronal uptake of the mediator are of lower physiological significance in younger animals as compared to those in adults. PMID- 3030030 TI - [The medium for determining the gelatinase activity of non-fermenting gram negative microorganisms]. AB - To determine the gelatinase activity of Gram-negative nonfermenting microorganisms, a double-layer medium containing 2% of agar in the lower layer and 10-20% of gelatin in the upper layer has been developed. The medium ensures free access of atmospheric oxygen, possibility of rapid (24-42 h) determination of gelatinase activity at 22-25 degrees C, efficacy of the test. simultaneous determination of gelatin hydrolysis by 8-12 strains in one dish. PMID- 3030031 TI - [Study of immunological mechanisms of action of temperature and emotional stress factors in experimental flavivirus infections]. AB - The influence of two stress factors, sharp changes in temperature and hypokinesia, on the course of experimental tick-borne encephalitis and Langat virus infections in mice has been studied. The data obtained in this study indicate that both factors produce defects in T- and B-cell-mediated immunity, accompanied by the activation of asymptomatic infection and the decrease of the mean survival time in acute infection. These two stress factors, differing in their intensity and nature (physical and emotional), have been shown to produce the same effect on the course of acute and asymptomatic flavivirus infections. In the former case the mean survival time of the animals decreases, and in the latter case clinically manifest infection develops. Under the conditions of hypokinesia (or changes in temperature), the death rate among the animals infected with langat virus has been found to increase 3- to 4-fold in comparison with the controls, the mortality level in the groups subjected to different stress factors being the same. PMID- 3030032 TI - Interaction of haemoglobin and cytochrome c with aliphatic alcohols as studied by 1H NMR spectroscopy. AB - Association of haeme proteins, haemoglobin and cytochrome c, with eight aliphatic alcohols (methanol, ethanol. two isomeric propanols and four butanols) was studied by 1H NMR spectroscopy. NMR spectra of alcohols were monitored at 60 MHz at increasing concentration of the proteins. Selective broadening of the NMR signals of individual segments of alcohols was observed only in the case of alcohol-haemoglobin systems. Its quantitative evaluation and interpretation in terms of formation of low affinity intermolecular alcohol--protein complexes led to the conclusion that haemoglobin associates with alcohol molecules in a way depending on the length and isomeric branching of the alkyl chains; in particular, the methylene and methine groups vicinal to the hydroxyl are subject to stronger immobilization than the terminal methyls or other groups. Thus, the model of hydrophobic complexes stabilized by hydrogen bonds described previously for association of bovine serum albumin with alcohols (Lubas et al., Biochemistry, 18, 4943-4951, 1979) seems to apply also to haemoglobin association. In the case of cytochrome c association, 1H NMR data alone are insufficient for structural evaluation of the mechanism of formation of the alcohol--cytochrome c complexes. PMID- 3030033 TI - Effects of conjugated estrogens on the vascular reactivity of the mesenteric vascular bed of the rat. PMID- 3030034 TI - A study of hypothalamic-pituitary-adrenal suppression following curative surgery for Cushing's syndrome due to adrenal adenoma. AB - The hypothalamic-pituitary-adrenal axis was investigated in all six patients requiring glucocorticoid replacement 2.5-11 years after unilateral adrenalectomy for adrenal adenomas causing Cushing's syndrome. The hypothalamic-pituitary adrenal axis was assessed by insulin induced hypoglycaemia and CRF testing in each patient. Two patients showed normal cortisol and ACTH responses to hypoglycaemia. Two patients showed subnormal cortisol responses to hypoglycaemia in the presence of high or normal basal ACTH concentrations. ACTH concentrations increased with both hypoglycaemia and CRF. Two patients showed subnormal cortisol responses to hypoglycaemia and CRF. One of these patients showed an ACTH rise following hypoglycaemia but not CRF. Defects at either hypothalamic-pituitary or adrenal levels were demonstrated and recovery of the axis appears to commence at the hypothalamic-pituitary level. PMID- 3030035 TI - The effect of submaxillariectomy on the uterine peroxidase activity and [14C]phenylalanine incorporation into uterine protein in rats. AB - Surgical removal of submaxillary glands in immature female rats caused an increase in size and about 3-fold increase in dry and wet weight of the uterus compared to that of the sham operated animals of the same age group. Histological examination reveal a significant increase in the diameter of the uterus with considerable elongation of the luminal epithelium from cubical to columnar in the experimental group. Biochemical studies showed that the uterine peroxidase, a marker enzyme for oestrogen action, increased (P less than 0.01) on submaxillariectomy. Incorporation of [14C]phenylalanine into the nuclear fraction of uterus was also enhanced significantly on removal of submaxillary glands. The elevation of peroxidase activity as well as of [14C]phenylalanine incorporation into the nuclear fraction due to removal of submaxillary glands were abolished in ovariectomized rats suggesting the involvement of ovarian hormones. The results show that oestrogen was responsible for all the above mentioned changes, whereas progesterone had little effect. Results further suggest the existence of a factor in the submaxillary glands through which they exert an effect on the uterus and ovary. PMID- 3030036 TI - Interaction of verapamil with d-tubocurarine and cholinergic agonists at the avian neuromuscular junction. AB - The effect of verapamil on neuromuscular transmission and muscle contraction was studied in the skeletal muscle of chick in vitro. The interactions of verapamil with d-tubocurarine (d-TC)-induced neuromuscular blockade, acetylcholine (ACh) and tetraethylammonium (TEA)-induced contractures were also studied. The purpose of the present investigation was to see if verapamil: intensified the neuromuscular blockade produced by d-TC; modified the cholinergic responses to ACh, TEA; and inhibited both directly and indirectly elicited twitch contractions. The results showed that verapamil (1-100 micrograms/ml, 2-200 mumol/l) had a neuromuscular blocking activity on its own; i.e. it reduced both directly and indirectly evoked twitch contractions, and intensified the neuromuscular blockade produced by d-TC. In addition, verapamil reduced the contractures produced by ACh and TEA in the chick muscle. The results are in favour of the possibility that verapamil acts by a mixture of pre- and post junctional effects at the chick neuromuscular junction. PMID- 3030037 TI - Studies on hypothalamo-pituitary corticoliberin system. I. Localization of corticotropin-releasing factor (CRF)- and neurophysin-immunoreactive neurons in the Mongolian gerbil (Meriones unguiculatus). AB - The immunohistochemical localization of CRF- and neurophysin-containing neurons in the hypothalamus of the Mongolian gerbil was studied by means of the PAP technique. The CRF-immunoreactive fibers were detected mainly in the outer layer of the median eminence of intact adult male gerbils. The CRF-positive neurons respond to aminoglutethimide (Elipten, Ciba) administration by showing increased immunoreactivity and an increase in the number of stained cell bodies in the parvocellular division of the paraventricular nucleus. Aminoglutethimide treatment results also in an increase in the number of neurophysin-immunoreactive nervous fibers localized in the internal layer of the median eminence. However, CRF-immunoreactive fibers are observed mainly in the outer layer of the median eminence while neurophysin-immunopositive axons are seen predominantly in the internal layer of this region. Since the axons of paraventricular neurons run to the median eminence and their staining ability is changed due to aminoglutethimide, their involvement in the endocrine control of hypophysial ACTH release is postulated. PMID- 3030038 TI - Neurological complications and concomitants of AIDS. AB - A survey of the literature of neurological manifestations associated with the acquired immune deficiency syndrome (AIDS) shows a broad disease spectrum affecting approximately one third of the patients in large hospital series. The complications include focal cerebral lesions caused by abscesses, lymphomas, leucoencephalopathy or infarcts as well as encephalitis, meningitis and myelitis. Most opportunistic infections of the central nervous system presumably are caused by toxoplasma gondii, cytomegalovirus and cryptococcus neoformans. One tenth of all patients have neurological disease as their initial symptom of AIDS. The diagnosis should always be considered in patients at risk and in males with an unusual neurological history or with a peculiar CT scan of the brain. Besides the opportunistic complications of AIDS, LAV/HTLV-III itself probably attacks the nervous system and gives rise to concomitant lesions of the long tracts of the spinal cord with ataxia, paresis and spasticity and to subacute encephalopathy and peripheral nerve abnormalities as well. PMID- 3030039 TI - The difference between non-selective and beta 1-selective beta-blockers in their effect on platelet function in migraine patients. AB - Platelet function has been postulated as playing a role in the pathogenesis of migraine. This study aimed to investigate to what extent beta 1-selective and non selective beta-blockers interfere with the platelet function in migraine patients. Twelve patients with classical migraine were included. After a 2-week drug-free period, the patients were randomly allocated to either beta 1-selective metoprolol (50 mg b.i.d.) or non-selective propranolol (40 mg b.i.d.) treatment for one month. After a wash-out period, the patients were changed to the corresponding beta-blocker for one month. ADP-induced platelet aggregability, platelet cAMP, ATP and ADP levels, plasma cAMP and TxB2 concentration, as well as the serum production of TxB2 were measured before and after each treatment period. After propranolol treatment, the patients showed lower ADP threshold values for producing irreversible platelet aggregation and lower platelet and plasma cAMP levels as compared to metoprolol. Neither of the beta-blockers induced any change in the plasma concentration or serum production of TxB2. In conclusion, non-selective beta-blockade (propranolol) significantly increases the platelet aggregability compared to beta 1-selective blockade (metoprolol). PMID- 3030040 TI - Intravenous injection of horseradish peroxidase in the rat stimulates corticosterone and adrenocorticotropic hormone release. AB - Wistar rats were given intravenous (i.v.) horseradish peroxidase (HRP) in saline in doses commonly used to study vascular permeability and the blood-brain barrier. Samples of blood were taken from conscious animals via indwelling catheters at intervals up to 6 h after the HRP injection. Plasma concentrations of adrenocorticotropic hormone (ACTH) and corticosterone were determined and compared with levels in control rats injected with saline alone. Rats injected with saline only presented levels of hormones within the low limits of normal indicating an insignificant influence of stress induced by the experimental procedure. Within 30 min of the i.v. HRP injection, the plasma concentrations of both ACTH and corticosterone increased to very high levels and remained so throughout the period of observation, namely 6 h. The time course of the changes in the concentrations was the same for the two hormones and the actual numerical values were related to the dose of HRP injected. The i.v. injection of HRP in Wistar rats, therefore, induces a marked release of stress hormones which by themselves have profound physiological effects. This phenomenon must be taken into account, in studies on normal vascular permeability using HRP as a tracer and also in similar experiments exploring various pathological conditions of the blood-brain barrier. PMID- 3030041 TI - Peripheral neuropathy in course of progressive systemic sclerosis. Light and ultrastructural study. AB - Progressive systemic sclerosis (PSS) is a chronic inflammatory disease of the connective tissue with involvement of the skin and other organs. The disease is characterized by an abnormal accumulation of collagen in all tissues and by microangiopathy. The involvement of the peripheral nervous system during PSS is very unusual and few cases are reported in the literature. A morphological study on the neuropathy associated with sclerodermia has been performed in rare cases. In this paper we demonstrate the role that the vascular lesions have in the pathogenesis of neuropathy during scleroderma. In particular, the primary role of the peripheral microangiopathy during PSS (observed in different clinical cases) is verified. PMID- 3030042 TI - Ultrastructural localization of Thorotrast (thorium dioxide) in human liver by analytical scanning electron microscope. AB - The distribution of thorium dioxide (Thorotrast) in the liver from a Thorotrast administered autopsy case was examined stereographically by analytical scanning electron microscopy. Thorium particles were detected in the macrophages of the portal triad and hepatic sinusoid. These macrophages were irregularly shaped and tended to be aggregated. In the sinusoid, accumulation of the macrophages formed a thrombus-like structure. Furthermore, observed in the sinusoid were free Thorotrast particles that appeared to have been released into the sinusoid as a result of breakdown of the macrophages (Kupffer cells). PMID- 3030043 TI - Calcitonin-producing insulinoma. An immunohistochemical and electron microscopic study. AB - A pancreatic endocrine tumor measuring 1.5 X 1.0 X 1.0 cm in a 67-year-old man with recurrent hypoglycemic attacks was examined by immunohistochemistry and electron microscopy. The tumor cells were relatively uniform, and almost all of them were immunoreactive to insulin. A few calcitonin-positive cells could be identified scattering in tumor cell strands, and the content of calcitonin in the tumor was 85 ng/g wet tissue. Coexistence of insulin and calcitonin in tumor cells was demonstrated by using adjacent semithin sections of the Epon-embedded material. Electron microscopically, the tumor consisted of a single type of cells with secretory granules identical to B-cell granules in the normal counterpart. PMID- 3030044 TI - Malignant fibrous histiocytoma showing cytoplasmic hyaline globules and stromal osteoids. A case report with light and electron microscopic, histochemical, and immunohistochemical study. AB - A 56-year-old man died of widespread metastases of malignant fibrous histiocytoma (MFH) primarily developed in the left lower gingiva after a rapid clinical course is reported. Primary and metastatic tumors showed light and electron microscopic, histochemical and immunohistochemical characteristics of MFH together with cytoplasmic hyaline globules and stromal osteoids. Hyaline globules revealed periodic acid-Schiff positive and diastase resistant reactions but not positive with any histochemical and immunohistochemical reactions. Osteoids were partially calcified and rimmed by osteoblastic tumor cells. The present case of MFH may provide difficulties in the differential diagnosis from some osteoid-forming sarcomas in soft tissues with relation to the histogenesis. PMID- 3030045 TI - Biochemical compartmentation of gluconeogenic enzymes in fish tissues. AB - Compartmentation of liver, kidney muscle and gill tissues in relation to glucose 6-phosphatase and fructose 1,6-diphosphatase was examined in the fishes Labeo rohita, Clarias batrachus and Channa punctatus. The anterior region of the right and left lobes of the liver contained the maximum of fructose 1,6-diphosphatase and glucose-6-phosphatase, while the minimum was in the right and left lobes of gill tissue. Herbivore fish had the highest gluconeogenic enzyme content followed by carnivore and piscivore species. The observed enzymatic variations in the three fish species were discussed. PMID- 3030047 TI - Effects of arginine-vasopressin and ACTH 4-10 on acquisition of active avoidance response in rats with alcohol pretreatment in prenatal and adult age. AB - Alcohol administration in drinking water to mother rats during gestation resulted in a permanent learning deficit of the offspring when behavioural reactions were tested in active avoidance conditioned reflex situation in adult age. A similar deficit of learning capacity was observed in adult rats following alcohol administration for two weeks; the acquisition of active avoidance response was tested later. Administration of arginine-vasopressin and ACTH 4-10 during behavioural test led to a significant improvement of learning ability of the animals pretreated with alcohol. The observations indicate that the ethanol induced deficit of learning capacity involves peptidergic mechanisms and the behavioural manifestations following chronic alcohol treatment are not irreversible processes. PMID- 3030046 TI - Effect on the plasma renin-aldosterone system of lesions to suprachiasmatic nuclei and central serotonin depletion in rats. AB - Renin activity, angiotensin-converting enzyme activity and aldosterone concentration were measured in the plasma of 8 experimental groups of rats: I- sham operated non-treated rats, II--suprachiasmatic nuclei (SCN) lesioned non treated: III--sham operated + furosemide (4 mg/kg i.p.), IV--SCN lesioned + furosemide, V--shams + 24-hour water deprivation: VI--SCN + 24-hour water deprivation, VII--intact rats + saline: and VIII--intact rats + p chlorophenylalanine (pCPA, 300 mg/kg, i.p.). No significant changes in basal levels of the three parameters were found after SCN, lesions in comparison with sham operated controls. Furosemide caused a similar increase in all three parameters of both sham and SCN lesioned rats. Similar changes were observed in SCN rats 24 hours after water deprivation and in intact rats 48 hours after serotonin depletion by pCPA: suppressed renin activity together with increased aldosterone concentration. It is concluded that the central serotonergic system and SCN play a similar role in control of the renin-aldosterone system in rats under conditions of negative water-salt balance. PMID- 3030048 TI - Effects of thyroid hormones and reverse-triiodothyronine (rT3) pretreatment on beta-adrenoreceptors in the rat heart. AB - Effects of thyroxine (T4), triiodothyronine (T3) or reverse-triiodothyronine (rT3) in vivo pretreatment or the effect of hypothyroidism on properties of beta adrenoreceptors in the rat heart were investigated. beta-adrenergic antagonist hydroxybenzylpindolol (HYP125I) was used to estimate the maximal binding capacity (MBC) and the affinity (KD) of beta-adrenoreceptors. Both T4 and T3 in dose and time dependent manner increased MBC value but only slightly and insignificantly affected affinity of the beta-adrenoreceptor. The effect of T3 on MBC was higher than that of an equal dose of T4, but the latter effect persisted longer than that of T3. Hypothyroidism decreased MBC value, but increased the affinity of beta-adrenoreceptors. Pretreatment of rats with rT3 produced changes in beta adrenoreceptors similar to those seen in hypothyroid rats. PMID- 3030049 TI - Breast cancer following multiple chest fluoroscopies among tuberculosis patients. A case-control study in Denmark. AB - A case-control study of breast cancer among tuberculosis (TB) patients in Denmark (1937-1954) was conducted to provide additional information on the radiation risk associated with low-dose chest fluoroscopy exposures. Records of 46013 TB patients were linked to the Danish Cancer Registry and 125 subsequent female breast cancers identified. Medical records were located for 89 (71%) of these women who developed breast cancer and on 390 controls, who were individually matched to cases on age and calendar year of TB diagnosis, and survival. Common risk factors for breast cancer such as nulliparity (relative risk (RR) = 2.5) and high relative weight (RR = 2.6) were also identified in this population of TB patients. However no risk was evident with exposure to any type of fluoroscopy (RR = 0.6; 95% CI = 0.2-1.4), or to fluoroscopies performed to monitor lung collapse therapy (RR = 0.8; 95% CI = 0.5-1.4). Although based on only 7 breast cancers, there was a suggestion of an increased risk among women who received greater than 1 Gy to their breasts (RR = 1.6; 95% CI = 0.4-6.3). Because of the infrequent use of fluoroscopy in our study, the breast doses were too low, 0.27 Gy on average, to expect to detect a significant elevation in breast cancer risk overall. The findings do suggest, however, that current estimates of breast cancer risk following radiation are not greater than presently accepted, and that a relative excess of 40 per cent can be excluded with reasonable confidence following breast doses on the order of 0.3 Gy. PMID- 3030050 TI - Sarcoma following irradiation for breast cancer. Report of three unusual cases including one malignant mesenchymoma of bone. AB - Three cases of sarcoma developing after irradiation for breast cancer are reported. A malignant mesenchymoma in the sternum--a combination of osteogenic sarcoma and rhabdomyosarcoma--is the first documented case of its kind occurring after radiation therapy. Of the other two tumors one was an extraskeletal osteogenic sarcoma in the soft tissues of the thoracic wall and one a rhabdomyosarcoma in the axilla. PMID- 3030051 TI - Survival and prognostic factors in thyroid carcinoma. AB - A multivariate analysis of prognostic factors and survival was carried out in a series of 200 patients with thyroid carcinoma. The cumulative survival rate corrected for intercurrent deaths was higher for papillary carcinoma than for follicular carcinoma both at 5 years (92% vs 74%) and at ten years (87% vs 66%) after the diagnosis. Seventeen of the eighteen patients with anaplastic carcinoma died within 24 months after the diagnosis. The most important independent prognostic factor in patients with papillary or follicular carcinoma, by multivariate analysis, was age at time of diagnosis, followed by tumor penetration beyond the thyroid capsule and follicular histologic type. When different types of treatment were included in the analysis, age at diagnosis still remained the most important prognostic factor. Misdiagnosed intercurrent deaths in the elderly did not explain the negative effect of age on survival. PMID- 3030053 TI - Effect of high 131I doses on the bone uptake and retention of 90Sr and 90Y. AB - The uptake and retention of 90Sr and 90Y in mouse bones after injections of the two nuclides in equilibrium were examined after heavy thyroid irradiations from 131I deposited in the glands. The radiation doses to the thyroid glands as well as the gross doses to the femurs and humeri of the mice were calculated. The radiation destruction of the thyroid tissues had no effect on the bone weights nor on the skeletal metabolism of 90Sr. The uptake of 90Y was, however, depressed after thyroidal irradiation but reached the same bone concentration as 90Sr at about 30 days after the administration of the nuclides, i.e. at a time when the corresponding equilibrium between 90Sr and 90Y in the bones was reached in mice without a thyroidal irradiation. PMID- 3030052 TI - Flow cytometric DNA patterns in cervical carcinoma. AB - Flow cytometric measurements of the DNA content were performed in a prospective study of 167 women with invasive squamous cell carcinoma of the uterine cervix. Ploidy level and the proportion of S-phase cells were correlated to age, menopausal age, staging according to FIGO and histopathology. With increasing age a successive shift from a dominance of peri-diploid and peri-tetraploid values to marked aneuploidy was found. Peri-diploid and peri-tetraploid tumors were more often found in premenopausal than in post-menopausal women (p less than 0.001). The mean S-phase rate was significantly higher in women aged 60-89 years than in women aged 20-59 years (p less than 0.01). More aneuploid tumors were found in stages III and IV than in stages IB and II (p less than 0.01). The mean S-phase rate was higher in stages III and IV (20.8%) than in stages IB and II (17.2%) (p less than 0.01). No statistically significant correlation was shown between ploidy level and histopathology or between S-phase rate and histopathology. In 37 patients polyclonal tumors were found. The reproducibility of the method was good (r = 0.99). PMID- 3030054 TI - Effect of high 131I doses to the thyroid gland on tumorigenicity of 90Sr and 90Y in mice. AB - The incidence of tumors was studied in mice injected with 90Sr only or with 90Sr in combination with high amounts of 131I. The high 131I-dose to the thyroid gland was necrotizing to the glandular tissue and the main aim of the investigation was the possible effects of the thyroidal destruction on the formation of bone tumors. After correction for competing mortality, no significant difference in the frequency of bone tumors could be found between 90Sr-treated and (90Sr + 131I)-treated mice. The incidence rate of bone tumors, however, was higher in mice with radiogenically destroyed glands than in those with intact glands. The limitations of using the concept of 'actuarial tumor incidence' in correction for competing mortality in animal experiments are discussed. Large numbers of lymphatic tumors were found in all animal groups. The frequencies of such tumors were independent of the radiation doses but their incidence rates were shortened in a dose dependent manner. Other, directly or indirectly radiation induced tumors were observed. PMID- 3030055 TI - Cell size following irradiation in relation to cell cycle. AB - The cell volume of Ehrlich ascites tumour cells was studied following a radiation dose of 5.0 Gy. The cell volume increased 12 to 30 hours after irradiation by about 20 per cent, was normal at about 50 hours, and increased again at 72 hours. In order to explain these changes the composition of the cells in cell cycle was studied. In addition, the cell volume of irradiated cells from the various parts of the cell cycle, separated by centrifugal elutriation, was measured. The changes in the mean cell volume of unseparated cells could be explained by variations in the cell cycle composition of the cell population. Irradiated cells from the various parts of the cell cycle did not deviate from the volume of non irradiated cells. The cell volume doubled during the cell cycle. This increase was, however, not linear. PMID- 3030057 TI - Radiosensitizing effects of nicotinamide on a C3H mouse mammary adenocarcinoma. A study on per os drug administration. AB - Nicotinamide is an inhibitor of adenosine diphosphate ribosyl transferase (ADPRT) which is involved in the mechanism of DNA repair after high doses of ionizing radiation. C3H mice with transplanted mammary adenocarcinomas were treated with low doses of nicotinamide, 10 mg/kg, 5 days a week, and in combination with ionizing radiation, 30 Gy, using different drug dose schedules. Mice given nicotinamide in combination with irradiation took a longer time to reach a tumor volume of 1,000 mm3 and a higher complete response rate (i.e. defined as total disappearance of the tumor for at least 7 days) than those given radiation alone. This was true whether nicotinamide was given daily from one week before tumor transplantation until the animal was killed or from transplantation day until day of irradiation. In addition, nicotinamide given per os at a dose between the recommended maximum daily allowance for human subjects (20 mg/70 kg), and the therapeutic allowance (1 g-12 g daily) 5 days a week for 9 weeks, showed a radiosensitizing effect without any histologically detectable damage to the normal tissues of the mouse, including bone marrow, intestine and the liver. PMID- 3030056 TI - Radiobiologic response of CHO-KI cells treated with vitamin A. AB - Treatment of CHO-KI cells with vitamin A altered their response to subsequent gamma irradiation. In general longterm preincubation with low doses of the vitamin caused a relative increase in the number of cells surviving a given radiation dose. The effect resulted in an increase in the D0 of the survival curve. Long or short term exposure to high concentrations of the vitamin caused a decrease in the number of surviving cells leading to a decrease in the extrapolation number of the survival curve. Recovery of cells from radiation damage, assessed using the split dose technique, was also impaired by vitamin A pretreatment. A mechanism involving repair of potentially lethal damage may explain the protective effect of vitamin A since this was highly dependent on the cell density of cultures at the time of irradiation. However, in view of the data showing that the vitamin A concentrations necessary to alter the radiation survival curve shoulder caused a significant release of sialic acid into the medium, a mechanism involving membrane stability may account for both the reduction in repair/recovery capacity of the treated cells and the radioprotective effect. PMID- 3030058 TI - Combined effects of ultrasound and ionizing radiation on lymphocyte chromosomes. AB - The effects of ionizing radiation and ultrasound upon the induction of chromosomal-type aberrations and sister chromatid exchanges were investigated. No statistically significant increase in the frequencies of dicentric and ring chromosomes or sister chromatid exchanges was discovered after ultrasound exposure alone at the diagnostic level. Nor could elevated frequencies of these phenomena be found following exposure to ultrasound before or after ionizing radiation, compared with those resulting from an equivalent dose of ionizing radiation alone. However, simultaneous exposure to ultrasound and ionizing radiation seemed to induce a slight enhancement of sister chromatid exchanges, although no significant change was noted in the yields of dicentric and ring chromosomes. PMID- 3030059 TI - Neutron dosimetry with detectors of finite size. I. Theory. AB - A theory for detector response in fields of fast neutrons has been developed. The theory is valid for convex detectors of all sizes exposed to isotropic fields of neutrons. In the theory the detector shape is characterized by distributions of chord lengths along which the neutron produced charged particles travel and deposit energy. Chord length distributions for charged particles generated in the surrounding medium (externals) and in the detector itself (internals) are presented for spheres, spheroids, cylinders and parallel plane geometries. The form on which the theory is written makes it possible to evaluate the theory with simplified assumptions, such as characterizing the detector with a single mean chord length for the external particles. Specifically, the response of ionization chambers has been considered, taking into account the energy dependence of the W values for the different charged particles. In a following article (part II) the theory will be subjected to experimental test. PMID- 3030060 TI - Neutron dosimetry with detectors of finite size. II. Experiments and calculations. AB - In a previous article a theory for detector response in fields of fast neutrons was presented. In the present paper this theory is subjected to experimental tests. For detectors of sizes comparable to the ranges of the neutron produced charged particles the theory predicts the variation of the detector response with detector size and elemental composition of the detector. A tissue-equivalent ionization chamber exposed to a field of 252Cf neutrons was used in the experimental tests of the theory. The size dependence of the response was investigated by varying the chamber gas pressure and the effects of different elemental compositions (mainly hydrogen content) was investigated by using different gases in the chamber (H2, CH4, TE-gas N2, Air, CO2, and Ar). The theory was evaluated both by using chord length distributions to characterize the chamber cavity and with a simplified version using a single mean chord length. The agreement between theory and experiment is generally good. PMID- 3030061 TI - [Nesidioblastosis and insulinoma. An infrequent association]. PMID- 3030062 TI - [Oncogenes]. PMID- 3030063 TI - Surgical and conservative treatment of cervical spondylotic radiculopathy and myelopathy. AB - One hundred and fourteen patients were admitted to our department for evaluation of their cervical spondylogenetic symptoms, including local cervical pain, radiculopathy and myelopathy. This retrospective study gives the results, expressed as improved, unchanged or worse, of anterior surgery, posterior surgery and conservative treatment. Local cervical pain improved in about half of the patients, without any difference between the groups. The effect of surgery on radiculopathy was superior to that of conservative treatment, 71 percent and 74 percent respectively, being improved after anterior and posterior surgery, compared to 19 percent in the conservatively treated group. The majority of patients with myelopathy were treated with posterior surgery and 69 percent had improved. The results were not influenced by the patients age or the duration of symptoms. It is argued that the positive effects of surgery on the radiculopathy are due to a segmental stabilisation rather then to decompression. The immediate post-operative improvement of the myelopathy is undoubtedly caused by the decompression while the long-termed improvement cannot with certainty be attributed to the operation. PMID- 3030064 TI - ["Burned out" testicular tumor]. PMID- 3030065 TI - Nonparathyroid hypercalcemia. PMID- 3030066 TI - Recent advances in diagnosis and treatment of pituitary tumors. PMID- 3030068 TI - Monoamine oxidase, hydrogen peroxide, and Parkinson's disease. PMID- 3030067 TI - Localization of MAO-A and MAO-B in human brain: a step in understanding the therapeutic action of L-deprenyl. AB - The discovery that MAO can be differentiated biochemically and pharmacologically into two forms, types A and B, with different substrate specificities and inhibitor sensitivities has renewed the interest in MAO inhibiting as a therapeutic agent. L-Deprenyl, a selective inhibitor of MAO-B, was introduced by us into clinical use as an adjunct to L-DOPA some years ago. This drug has found therapeutic importance in that it can potentiate the pharmacological action of L DOPA. The mechanism underlying the action of L-deprenyl is thought to be related to its inhibition of MAO-B and thus increased levels of PEA and DA, as measured in the striatal and limbic region of human brain. Animal studies have indicated the MAO-A is mainly, but not exclusively, located in brain neurons, while MAO-B is preferentially placed in glia and astrocytes. In general human and primate brain studies show similar MAO distribution. The observation that MAO-B activity could not be located in the catecholaminergic neurons of human brain by the use of monoclonal antibody studies seriously questions the validity of this technique. The exact locations of MAO-A and -B in human brain are important to understand the mechanism of L-deprenyl action as an antiparkinson drug. If there is an absence of MAO-B from dopaminergic neurons, one may now consider that MAO-B activity within glia plays a far more important role than hitherto considered. This, however, is questionable. PMID- 3030069 TI - Study of platelet alpha 2-adrenoceptors in Parkinson's disease. PMID- 3030070 TI - The Lewy body in Parkinson's disease. AB - In this brief overview three problems regarding Lewy bodies have been taken up for discussion: the specificity of the Lewy bodies, the extranigral sites for the inclusions, and the variable morphology and composition of the bodies in different portions of the nervous system. These questions go to the heart of the problem of nerve cell degeneration in Parkinson's disease. The question about the specificity of Lewy bodies has been answered in the affirmative. It therefore is worthwhile to examine the Lewy bodies, find out what they are composed of, and what molecular events precede and accompany their formation. Once we know that, will we be able to prevent Lewy bodies from forming? And if Lewy bodies do not form, will we then have no substantia nigra degeneration and no Parkinson's disease? Perhaps that is too much to expect from Lewy's peculiar cellular inclusions. PMID- 3030071 TI - Familial Alzheimer's disease presenting as levodopa-responsive parkinsonism. PMID- 3030072 TI - GABA receptor agonists and extrapyramidal motor function: therapeutic implications for Parkinson's disease. AB - GABA receptor agonists display a dual action on DA-mediated events. One includes a decrease in DA release, reduction in DA receptor density, and decreased response of postsynaptic cells to dopaminergic stimulation; it results in antidopaminergic effects. The other consists of a reduction of striatal cholinergic activity resulting in a facilitation of dopaminergic effects. These two effects could be dissociated depending on the dose of GABA receptor agonists. This dual action probably explains the results of clinical trials showing either amelioration of parkinsonian symptoms with aggravation of L-DOPA-induced dyskinesia or improvement of dyskinesia without or with aggravation of parkinsonian symptoms. PMID- 3030073 TI - Nigral and adrenal grafts in parkinsonism: recent basic and clinical studies. PMID- 3030074 TI - Hydroxyl radical scavenging action of tinoridine. AB - Radical scavenging action of tinoridine, a non-steroidal anti-inflammatory drug with a potent anti-peroxidative activity, was investigated. Tinoridine reduced a stable free radical, diphenyl-p-picryl-hydrazyl, in the molar ratio of about 1:2, indicating its free radical scavenging ability. Tinoridine inhibited the lipid peroxidation in rat liver microsomes induced by xanthine-xanthine oxidase system in the presence of ADP and Fe2+, in which hydroxyl radical (. OH) is formed. Tinoridine was demonstrated to be oxidized in the course of the lipid peroxidation by following the fluorescence derived from the oxidation product of tinoridine. It was also oxidized by the xanthine-xanthine oxidase system in the presence of Fe2+, but its oxidation was slow in the absence of Fe2+ and almost completely inhibited by catalase. Tinoridine was also oxidized by H2O2-Fe2+ system producing . OH (Fenton reaction), but it did not affect the reduction of cytochrome c caused by superoxide radical. These results indicate that tinoridine is able to scavenge . OH and the main active oxygen species responsible for the lipid peroxidation is . OH. The anti-peroxidative and . OH scavenging ability of tinoridine should contribute to its anti-inflammatory action. PMID- 3030075 TI - Prostaglandin E2 and collagenase release by cultured talus from type II collagen arthritis rats. PMID- 3030076 TI - A possible role for arachidonic acid metabolism in guinea pig tracheal beta adrenergic receptor modulation by pulmonary macrophages. PMID- 3030077 TI - Effects of dietary poly-unsaturated fatty acids on the beta-adrenergic receptor reactivity in the guinea pig respiratory system. PMID- 3030078 TI - Endotoxin-induced reduction of beta-adrenoceptor number in guinea pig splenic lymphocyte membranes. PMID- 3030079 TI - Endotoxin-induced alterations in guinea pig coronary vascular beta 2-adrenoceptor function: a role for cyclo-oxygenase products. PMID- 3030080 TI - [Epstein-Barr virus-specific antibody pattern in sarcoid uveitis with and without systemic manifestations]. PMID- 3030081 TI - [Morphological studies of filamentous inclusions in the human lens capsule]. PMID- 3030082 TI - MR imaging of hepatic focal nodular hyperplasia: characterization and distinction from primary malignant hepatic tumors. AB - Spin-echo MR imaging at 0.35 T was used to image hepatic focal nodular hyperplasia (FNH) and to attempt to distinguish it from primary malignant hepatic tumors. There were six FNH and 10 malignant tumors including seven hepatocellular carcinomas, two cholangiocarcinomas, and one hepatoblastoma. Our results show that FNH has a fairly consistent appearance, dissimilar from that of malignant primary hepatic tumors. Four of six FNH lesions were isointense (except for a central scar in three) and indistinguishable from normal hepatic parenchyma on all pulse sequences, whereas two of six were homogeneous but slightly hyperintense on T2-weighted sequences. Furthermore, a central hyperintense scar was seen in three of six lesions on T2-weighted sequences. In contrast, each of the malignant primary hepatic tumors was hyperintense on T2-weighted sequences and seven of 10 were hypointense on T1-weighted sequences; in nine of 10, heterogeneous areas of intensity were noted. In two fibrolamellar hepatocellular carcinomas a central scar was seen that was hypointense on all pulse sequences. By using quantitative data, the best characterization was obtained by using lesion/normal-liver intensity ratios from a T2-weighted sequence; all FNH had a ratio less than 1.33, while in nine of 10 primary malignant tumors it was greater than 1.41. We conclude that focal nodular hyperplasia may have a consistent appearance on spin-echo MR imaging and probably can be distinguished from primary malignant lesions in most instances. PMID- 3030083 TI - MR imaging of pituitary adenoma: CT, clinical, and surgical correlation. AB - Twenty-five patients with suspected pituitary adenoma were evaluated prospectively with CT and MR. Nine patients underwent transsphenoidal surgery, and three of these showed a documented decrease in size of mass on bromocriptine therapy. CT was more sensitive than MR for detecting focal lesions (seven vs three) and sellar-floor erosion (12 vs six). MR was superior to CT in identifying infundibular abnormalities (seven vs six), focal abnormalities of the diaphragma sellae (10 vs seven), cavernous sinus invasion (four vs two), and optic chiasm compression (six vs zero). Thus, MR may be the procedure of choice for optimal identification and localization of macroadenoma. For patients with suspected microadenoma, however, this preliminary series indicates that CT remains the radiographic procedure of choice. PMID- 3030084 TI - Transient phenytoin induced IgA deficiency and permanent IgE increase. AB - This is a report of a 9-year-old epileptic boy, who was studied over a period of 7 years. The seizures started when he was 2 months old. He was treated with phenytoin from the age of 2 years and 7 months. Serum and salivary IgA were absent with high IgE serum total. The routine immunologic studies were normal. The IgA was normalized after phenytoin withdrawal, but IgE determination increased progressively without any atopic symptoms. The T4 (helper)/T8 (suppressor) ratio decreased (1.0 and 1.2) on two different days, although above the normal limit. The phenytoin only modified the IgA levels. These data suggest that a primary immunoregulatory abnormality may be present in drug induced IgA deficiency. PMID- 3030085 TI - Drugs for protozoan infections. PMID- 3030086 TI - Early identification of amyloid heart disease by technetium-99m-pyrophosphate scintigraphy: a study with familial amyloid polyneuropathy. AB - To determine whether technetium-99m-pyrophosphate (Tc-99m-PYP) scanning or two dimensional echocardiography can detect amyloid heart disease in an earlier stage of familial amyloid polyneuropathy, 15 patients were examined. Although 10 of the 15 patients had no clinical evidence of congestive heart failure, as well as normal ventricular wall thickness and normal values for left ventricular systolic function, five (50%) of them showed mild or moderate myocardial uptake. On the other hand, none had characteristic highly refractile myocardial echoes on the two-dimensional echocardiographic images (p less than 0.01), and values for diastolic function were reduced in four of the five and normal in the remaining one. In 85 control subjects, diffuse positive pyrophosphate scans of the heart were found in four (5%) of them (three with dilated cardiomyopathy and one with sarcoidosis), and highly refractile granular sparkling echoes were observed in nine (11%) (five with hypertrophic cardiomyopathy, three with aortic stenosis, and one with hypereosinophilic syndrome). We conclude that Tc-99m-PYP scanning is a more sensitive and specific method and may have the potential ability to detect amyloid heart disease in the earlier stage of familial amyloid polyneuropathy than two-dimensional echocardiography. PMID- 3030087 TI - Comparison of enalapril and bendrofluazide for treatment of systemic hypertension. AB - The antihypertensive and biochemical effects of the angiotensin-converting enzyme inhibitor enalapril were compared with those of the thiazide diuretic bendrofluazide. Patients with untreated mild to moderate essential hypertension were entered into a randomized, double-blind, double-dummy, parallel-group study. Blood pressure (BP) decreased significantly (p less than 0.001) with either drug therapy: from 172/103 to 147/87 mm Hg (n = 18) with enalapril and from 179/104 to 165/95 mm Hg (n = 22) with bendrofluazide; thus, enalapril produced a greater reduction (p less than 0.05) than bendrofluazide. The median dose of enalapril was 20 mg/day (range 10 to 40) and that of bendrofluazide was 5 mg/day (range 2.5 to 10). Both drugs reduced serum sodium levels by small amounts. This was significant only for enalapril (decrease of 1.3 mmol/liter, p less than 0.05). Hyponatremia was not seen in any patient. Three patients were withdrawn from each treatment group due to adverse effects, although both drugs were generally well tolerated. PMID- 3030088 TI - Plasma beta-endorphin levels in silent myocardial ischemia induced by exercise. AB - Although silent myocardial ischemia is a well recognized phenomenon, the reasons for the lack of symptoms in patients with coronary artery disease (CAD) is unclear. Because the endogenous opioid beta-endorphin has been related to pain modulation, plasma beta-endorphin levels were studied before, during and after exercise-induced ischemia in symptomatic and asymptomatic men. Because beta endorphin responses have been closely linked to adrenocorticotropic hormone (ACTH) and cortisol responses, these hormones also were measured. Nine symptomatic and 12 asymptomatic patients with a high probability (at least 95%) of CAD and 8 apparently healthy men completed a Bruce protocol treadmill test. Blood samples were drawn before, during and 10 minutes after exercise. During exercise the measured hormones showed no significant increases from basal levels. However, plasma beta-endorphin, ACTH and cortisol levels were significantly elevated (p less than or equal to 0.01) 10 minutes after exercise in all 3 groups. There was no significant difference in plasma beta-endorphin levels during or after exercise between the symptomatic and asymptomatic patients with CAD. Thus, differences in circulating levels of beta-endorphin, ACTH and cortisol are not associated with the presence or absence of pain during exercise-induced myocardial ischemia. PMID- 3030089 TI - Cellular toxicity of fecal water depends on diet. AB - To determine whether concentrations of potentially toxic lipids in the aqueous phase of human stool are responsive to changes in dietary fat, calcium, and fiber, 20 male volunteers were placed on a high-fat, low-calcium, low-fiber or a low-fat, high-calcium, high-fiber diet for 4 days. To assess toxicity of the fecal fractions, we examined the ability of fecal supernatants to lyse human erythrocytes. Bile acid concentrations in fecal water from the low-fat group were reduced significantly from 180 +/- 60 microM to 100 +/- 70 microM; in the high fat group, increased from 190 +/- 60 microM to 250 +/- 100 microM. Erythrocyte lysis was 76% for the high-fat group, 37% for the low-fat group. There was a significant weak correlation between aqueous bile acid concentration and cell lysis. Results suggest that diet can influence concentrations of detergents in the aqueous phase of human stool and the potential toxicity of this fraction to cell membranes. PMID- 3030090 TI - Effects of graded sucrose additions on taste preference, acceptability, glycemic index, and insulin response to butter beans. AB - Dried beans, because of their high-fiber content and low-glycemic index, are especially suitable for diabetic diets. Most South African bean recipes contain sucrose, and since a restriction of artificial sweeteners seems desirable, replacing sucrose would be impractical. Hence, we examined the effects of 10, 20, and 30% sucrose additions to cooked dried butter beans on taste preference and acceptability in 29 diabetic patients and 11 control subjects. The effect of sucrose additions on glycemic index and insulin response to butter beans was determined in control subjects. Both diabetic and control subjects preferred beans with sucrose additions (p less than 0.005). Additions of sucrose up to 20% of total carbohydrate had no adverse effects on glycemic index or insulin response of butter beans (p less than 0.05), which indicates that addition of moderate amounts of sucrose to a low glycemic index food may improve palatability without impairing the favorable effect on blood glucose and insulin response. PMID- 3030091 TI - Primary oat cell carcinoma of the larynx. AB - The aggressiveness of small (oat) cell carcinoma of the larynx presents a therapeutic challenge to the oncologist. Since the first description of this type of carcinoma in 1972, 52 patients have been reported in the literature and a variety of treatment regimens have been used. The purpose of this study was to report two new cases and review all previous reports to determine the disease's biological behavior, clinical manifestations, and optimum treatment. Thirty-five percent of the tumors were transglottic, and 27% were supraglottic. Fifty-four percent of patients had regional metastases at initial presentation and 17.6% had distant metastases. The median survival was 10 months for all patients. Patients who were treated with chemotherapy with or without other modalities had the best 2-year survival rates (52.2%). Forty-one percent of patients had regional recurrence only, 12.5% had regional recurrence and distant metastases, and 2% developed distant metastases only. We conclude that patients with oat cell carcinoma of the larynx should be treated with combination chemotherapy and radiation therapy. Surgery is best reserved for persistent and recurrent disease at the primary site and neck. PMID- 3030092 TI - Cyclophosphamide, etoposide, and infusion cisplatin in refractory small cell lung cancer. A preliminary report. AB - Twelve patients (three with limited and nine with extensive disease) with small cell carcinoma of the lung who had plateaued or progressed on their chemotherapeutic regimens were switched to a program of combination chemotherapy consisting of cyclophosphamide, etoposide, and a 5-day continuous infusion of cisplatin. Nine of the 12 patients (75%) showed further tumor reduction, including four patients who achieved a clinical complete response. Also noted was a patient who previously failed on a regimen of short infusion cisplatin who responded to the continuous infusion platinum regimen. Toxicity was quite acceptable, considering the amount of prior treatment given these patients. Further studies seem warranted to confirm these encouraging preliminary results. PMID- 3030093 TI - Extracellular hyaline bodies are basement membrane accumulations. AB - Diverse hyaline bodies occurring in various diseases were investigated immunocytochemically with antibodies to type IV collagen and laminin. These hyaline bodies included those found predominantly extracellularly in adenoid cystic carcinomas, endodermal sinus tumors, Spitz nevi, lichen planus, diabetic glomerular nodular sclerosis, and odontogenic cysts, and those found predominantly intracellularly in alcoholic liver disease, Kaposi's sarcoma, and malignant fibrous histiocytoma. The hyaline bodies found in the first six diseases displayed intense immunoreactivity for the basement membrane components, type IV collagen, and laminin, whereas the hyaline bodies occurring in the latter three diseases were unreactive. These findings suggest that a common histogenesis of the extracellular hyaline body is the stimulatory effect of an altered growth state (hyperplasia, neoplasia, or endocrinopathy) on basement membrane synthesis in cells that normally produce a basement membrane. PMID- 3030094 TI - Appendectomy in South African inter-ethnic school pupils. AB - In 1984-1985, prevalences of appendectomy in inter-ethnic series of South African school pupils of 16-18 yr were: rural blacks, 0.5%; urban blacks, 1.0%; Indians, 2.6%; coloreds (Eur-African-Malay), 2.2%; Afrikaans whites, 13.4%; and English whites, 9.9%. Corresponding respective annual incidences per 1000 pupils of 10-19 yr were: 0.3, 0.6, 1.9, 1.7, 9.8, and 7.8. Thus, appendectomy is rare or infrequent in all except the white populations. Peak occurrence was postpubertal. There was no consistent sex bias. Dietarily, mean daily fiber intake was relatively low in all groups, 17.9-26.1 g. While the percentage of energy from fat intake was low in blacks, 16.3-22.3%, it was much higher in the other populations, 32.7-39.5%. Clearly, factors other than diet are involved in regulating frequency of appendectomy. While mortality is negligible and morbidity slight, elucidation of causation and prevention of the disease is desirable since subsequently appendectomy patients are at greater than average risk to certain cancers. PMID- 3030095 TI - Platelet dysfunction associated with Wilms tumor and hyaluronic acid. AB - Acquired von Willebrand disease (AVWD) has been described in two cases of nephroblastoma. We studied a patient with nephroblastoma who presented with a coagulopathy suggestive of AVWD. The subject had undetectable levels of F.VIIIR:Ag, diminished F.VIIIR:WF (16.3%), F.VIII:C activity (37%), and lack of platelet aggregation to ADP, epinephrine, collagen, and arachidonic acid. These results were associated with abnormally high serum levels (850 mg/dl) of hyaluronic acid (HA), which made the patient's serum hyperviscous. Examination of the neoplasm revealed HA in the tumor matrix. All abnormalities of coagulation resolved after chemotherapy and extirpation of the neoplasm, which produced normal serum HA levels. To study the effects of HA on platelet function, we added HA to normal platelet-poor plasma (NPP), which rendered F.VIIIR:Ag undetectable; treatment of HA with hyaluronidase eliminated F.VIIIR:Ag assay interference caused by HA. F.VIII:C activity decreased in vitro when HA was mixed with normal platelet-poor plasma (NPP). HA reduced the initial slope of normal platelet aggregation. Partial correction of platelet aggregation occurred after hyaluronidase treatment of HA-spiked PRP. Experiments in rabbits exposed to HA (serum level 400 mg/dl) demonstrated abnormalities similar to those noted in the patient. Shear rate studies of whole blood containing HA (500 mg/dl) yielded high shear stress, 27-136 dynes/cm2 over shear rates of 10-216 sec-1. We conclude that the coagulopathy demonstrated in this case is secondary to hyperviscosity produced by elevated levels of HA, which interferes with the assay for F.VIIIR:Ag. Thus the acquired coagulopathy associated with other cases of nephroblastoma may present as spurious von Willebrand disease. PMID- 3030096 TI - Correction of hypokalemia in Bartter's syndrome by enalapril. AB - Seven patients with Bartter's syndrome were investigated before and after 3 months' treatment by enalapril. Serum potassium rose from 2.4 +/- 0.5 to 3.9 +/- 0.6 mmol/L. In all patients, serum magnesium rose and bicarbonate fell. Hormonal changes were as suspected: a further stimulation of renin and a decline in aldosterone. The BP sensitivity to angiotensin II normalized in the five patients in whom the test was performed. Clearance studies during maximal water diuresis, performed in four patients, were compatible with a high proximal fractional tubular sodium reabsorption and a relatively low distal fractional sodium reabsorption. Fractional free water excretion after furosemide was also low, confirming the concept of a primary sodium reabsorption defect in the furosemide insensitive part of the nephron in Bartter's syndrome. The only consistent change after enalapril was a further decline in distal fractional sodium reabsorption. Initiation of therapy produced a BP fall in each subject. Clinical important hypotension associated with oliguria was seen twice, but these reactions were short-lasting. The BP rose to pretreatment values within 72 hours, despite continuation of converting-enzyme inhibition. Renal function recovered, though a moderate fall in function persisted. No other side effects were noticed. We conclude that converting-enzyme inhibition improves the potassium metabolism of patients with Bartter's syndrome, without ameliorating the abnormal renal sodium handling. PMID- 3030097 TI - Silica and glomerulonephritis: case report and review of the literature. AB - A 54-year-old foundry worker with extensive silica exposure, but no pulmonary disease, developed the nephrotic syndrome and renal failure over a 3-month period. Renal biopsy demonstrated a proliferative glomerulonephritis; energy dispersive x-ray analysis detected silicon within the renal tubules. Measurements of respirable silica at the foundry revealed levels up to 2.5 times the current occupational standard. Similar glomerular disease has been reported in silica exposed animals and workers with silicosis. This case suggests that clinicians should include silica exposure in the differential diagnosis of unexplained diffuse proliferative glomerulonephritis, renal disease may occur without clinically evident pulmonary disease in silica exposure, and silica-induced glomerulonephritis warrants further clinical and epidemiologic research. PMID- 3030098 TI - Neoplastic hypercalcemia: physiologic response to intravenous etidronate disodium. AB - Following a four-day control period during which an elevated serum calcium level either stabilized or continued to rise despite maximally tolerated saline diuresis, 12 patients with neoplastic hypercalcemia were treated with intravenous etidronate disodium (etidronate) 7.5 mg/kg/day for up to seven days. Serum calcium reverted to normal levels in all patients, with the mean pretreatment serum calcium level of 12.5 +/- 0.4 mg/dl dropping to 9.2 +/- 0.2 mg/dl (p less than 0.01) by Day 7. Elevated urinary calcium (1,107 +/- 134 mg/g creatinine) and hydroxyproline levels (154 +/- 16 mg/g creatinine) declined to 245 +/- 52 mg/g creatinine and 75 +/- 14 mg/g creatinine, respectively, suggesting a marked reduction in bone resorption following treatment. Serum phosphorus levels were unchanged, but urinary phosphorus levels dropped rapidly from 1,181 +/- 125 mg/g creatinine before treatment to 723 +/- 94 mg/g creatinine after two days. Serum parathyroid hormone levels (mid-molecule assay) were suppressed before treatment (64 +/- 16 pg/ml), but rose rapidly to 223 +/- 68 pg/ml by Day 7 of treatment. The value of serum 1,25-dihydroxyvitamin D was initially below normal (16 +/- 3 pg/ml), but rose rapidly with treatment to 42 +/- 12 pg/ml by Day 7. Symptoms of hypercalcemia and bone pain improved with treatment, and no serious adverse reactions to treatment were encountered. Intravenous etidronate is apparently an effective and safe treatment for neoplastic hypercalcemia. PMID- 3030099 TI - Glucagonoma syndrome. Rapid response following arterial embolization of glucagonoma metastatic to the liver. AB - Transcatheter arterial embolization was used to treat a patient with glucagonoma metastatic to the liver after chemotherapy failed. Rapid amelioration of the syndrome's manifestations followed. PMID- 3030100 TI - Angioneurotic edema, agranulocytosis, and fatal septicemia following captopril therapy. PMID- 3030101 TI - Cytomegalovirus infection in homosexual men. Relationship to sexual practices, antibody to human immunodeficiency virus, and cell-mediated immunity. AB - To investigate the relationships among cytomegalovirus infection, sexual behavior, human immunodeficiency virus (HIV) seropositivity, and indexes of cellular immunity, 180 homosexual men and 26 heterosexual men were studied. Among the homosexual men, cytomegalovirus seropositivity was associated with increased T8 lymphocyte counts (p less than 0.001) and reduced T4/T8 ratios (p = 0.006); these results were independent of HIV infection. Cytomegalovirus seropositivity was also associated with increasing age, numbers of sexual partners, and the practice of anal-receptive intercourse. At the first visit, cytomegalovirus was isolated from none of 13 cytomegalovirus-seropositive heterosexual subjects, compared with 62 (36 percent) of 171 seropositive homosexual men (p less than 0.005). Viral isolation was most common from semen. Among 32 cytomegalovirus seropositive homosexual subjects from whom culture specimens were obtained more than four times over 10 to 30 months, 72 percent of the specimens were culture positive. The mean duration of cytomegalovirus excretion in semen was 22 months, and in urine, the duration was nine months. Cytomegalovirus excretion was associated with younger age and reduced lymphocyte proliferation in response to cytomegalovirus, but not with antibody to HIV. Cytomegalovirus infection is sexually transmitted among homosexual men, perhaps by rectal intercourse, and is associated with alterations in T lymphocyte subsets. Most seropositive homosexual men excrete cytomegalovirus intermittently, primarily in the semen. PMID- 3030103 TI - Renal excretion of calcium in human hypertension. PMID- 3030102 TI - HTLV-I-associated lymphoma involving the entire alimentary tract and presenting with an acquired immune deficiency. AB - A patient with HTLV-I-associated adult T cell lymphoma presented like a patient with the acquired immune deficiency syndrome (AIDS) with peripheral lymphadenopathy, profuse diarrhea, weight loss, night sweats, and multiple opportunistic infections. Infections included cytomegalovirus, Pneumocystis, Histoplasma, Strongyloides, Giardia, and Staphylococcus aureus. Histoplasma has not previously been associated with this lymphoma. The lymphoma involved the entire alimentary tract from pharynx to rectum, which has not previously been reported for adult T cell lymphoma, and is rare for any lymphoma, with only one case reported in this century. Despite extensive gastrointestinal involvement, steatorrhea was not present, perhaps due to preservation of normal intestinal villous architecture. PMID- 3030104 TI - Altered cAMP content in specific brain areas of spontaneously hypertensive rats dependent on calcium status or parathyroidectomy. PMID- 3030105 TI - Aldosterone, calcium, and hypertension. PMID- 3030106 TI - When and how do you use a Heimlich flutter valve? PMID- 3030107 TI - Uterine sarcoma: an analysis of 74 cases. AB - In order to determine whether recent methods of diagnosis and treatment have resulted in an improved survival for patients with uterine sarcoma, we reviewed 99 cases treated in our center from 1970-1985. Morphologic characteristics of 74 tumors were specifically reassessed for this study. All tumors were graded. Of 42 Stage I cases that were morphologically assessed, tumor-positive pelvic lymph nodes were found in two of the 15 patients in whom sampling was done. No cases of tumor-positive para-aortic nodes were found in 14 patients with Stage I disease. In Stage I and Stage II, no cases of positive para-aortic nodes were found in association with negative pelvic nodes. The 2- and 5-year survival rates in Stage I were 47.4% and 29.4%, respectively. Local recurrence decreased (p less than 0.01) in Stage I from nine of 22 cases in which operation alone was performed to none of 15 cases in which pelvic radiotherapy was added, but no improvement in the 5-year survival rate was observed. As with lymphadenectomy and radiotherapy, the recent use of chemotherapy for uterine sarcoma had no impact on survival. PMID- 3030108 TI - Effects of adenosine 3',5'-cyclic monophosphate, forskolin, and cholera toxin on hormone production in human term placental cells. AB - Trophoblast cells from human term placenta in monolayer culture were used to investigate the role of adenosine 3',5'-cyclic monophosphate in the production of human chorionic gonadotropin and estradiol-17 beta. The intracellular concentration of adenosine 3',5'-cyclic monophosphate was elevated by (1) addition of an adenosine 3',5'-cyclic monophosphate analogue, 8-bromoadenosine 3',5'-cyclic monophosphate, (2) inhibition of the hydrolysis of Gs-GTP complex by cholera toxin, and (3) direct stimulation of adenylate cyclase with forskolin. Addition of 2 mmol/L of 8-bromoadenosine 3',5'-cyclic monophosphate markedly increased the accumulation of human chorionic gonadotropin in the culture medium on days 2, 3, and 4 of treatment. Likewise, addition of cholera toxin (0.2 microgram/ml) or forskolin (50 mumol/L) also enhanced human chorionic gonadotropin production. On the other hand, the production of estradiol-17 beta was significantly inhibited by 8-bromoadenosine 3',5'-cyclic monophosphate, at the same time that human chorionic gonadotropin production was enhanced in the same experiments. These results further support a differential role of adenosine 3',5'-cyclic monophosphate on human chorionic gonadotropin and estradiol-17 beta production in the human term placenta. PMID- 3030109 TI - Ampicillin/sulbactam versus metronidazole-gentamicin in the treatment of soft tissue pelvic infections. AB - The clinical efficacy and safety of ampicillin/sulbactam versus metronidazole gentamicin were evaluated in a comparative, randomized, prospective study. Forty four patients were enrolled: 22 received the ampicillin/sulbactam regimen, and 22 received the metronidazole-gentamicin combination. There were 33 cases of severe acute pelvic inflammatory disease, two tuboovarian abscesses, five cases of endomyometritis, and two cases of posthysterectomy pelvic cellulitis. Aerobic and anaerobic cultures from the infection sites yielded 447 microorganisms from 44 patients (an average of 10 bacteria per infection; 6.4 anaerobes and 3.7 aerobes). The most frequent isolates were Bacteroides sp., 54; Bacteroides bivius, 17; black-pigmented Bacteroides, 12; Bacteroides disiens, 11; Fusobacterium, 13; Peptostreptococcus anaerobius, 24; Peptostreptococcus asaccharolyticus, 21; anaerobic gram-positive cocci, 34; Gardnerella vaginalis, 29; Neisseria gonorrhoeae, 17; alpha-hemolytic streptococci, 15; and Escherichia coli, five. Clinical cure was noted in 19 of 20 patients treated with ampicillin/sulbactam and 18 of 21 patients treated with metronidazole-gentamicin. One treatment failure occurred in the ampicillin/sulbactam group in a patient who required antichlamydial therapy and had a complex left adnexal mass consistent with an abscess. The cases of metronidazole-gentamicin failure included two patients initially diagnosed as having tuboovarian abscesses who required a change in antibiotic therapy to control the infections. The third patient had postabortion endomyometritis that did not respond to metronidazole-gentamicin therapy within 48 hours, and required a change of medication. No adverse hematologic, renal, or hepatic effects were noted in either group of patients. PMID- 3030110 TI - The effects of human chorionic gonadotropin on growth and steroidogenesis of the human fetal adrenal gland in vitro. AB - This study examined the effects of human chorionic gonadotropin, adrenocorticotropin, human luteinizing hormone, and mouse epidermal growth factor on growth (thymidine incorporation) and steroidogenesis (dehydroepiandrosterone sulfate production) of human fetal zone adrenal cells in monolayer culture. Two preparations of human chorionic gonadotropin extracted from pregnancy urine were used, one highly purified (National Institutes of Health, CR-121) and one less pure (Sigma). Thymidine incorporation was increased twofold to tenfold in cultures exposed to the Sigma human chorionic gonadotropin preparation or to mouse epidermal growth factor as compared to control. Pure National Institutes of Health human chorionic gonadotropin and luteinizing hormone had no effect on growth. When adrenocorticotropin was added alone or in combination with Sigma human chorionic gonadotropin or mouse epidermal growth factor, growth was decreased. Dehydroepiandrosterone sulfate production was stimulated by adrenocorticotropin but not by human luteinizing hormone, human chorionic gonadotropin, or mouse epidermal growth factor. These results suggest that human pregnancy urine contains a growth factor which remains to be identified but that pure human chorionic gonadotropin has no mitogenic or steroidogenic effects on the cultured fetal zone cells of the human fetal adrenal gland. PMID- 3030111 TI - Interferon-alpha therapy of recurrent conjunctival papillomas. AB - Five patients with multiple, recurrent conjunctival papillomas underwent surgical excision of their tumor and then received interferon alpha-N1, 5 X 10(6) units/m2 (5 Mu/m2), intramuscularly daily for one month. A similar dose was given two to three times per week for the next six months and tapered off or discontinued thereafter. The follow-up period varied from one to four years. Two patients have had no recurrence of tumor. The other three patients have had recurrences of lesser severity upon tapering or discontinuing the interferon, and repeat surgical or laser excision of these lesions has been performed. The presence of koilocytosis and human papillomavirus type 11a DNA sequences was noted in all specimens large enough to examine, whereas papillomavirus structural antigens were detected in only two of five specimens. A regimen of interferon therapy appears to be tumor suppressive, but not curative. PMID- 3030112 TI - Cytomegalovirus retinitis as the initial manifestation of the acquired immune deficiency syndrome. AB - We studied eight patients in whom cytomegalovirus retinitis was an initial manifestation of AIDS. Six of these patients had cytomegalovirus retinitis alone at the time of diagnosis and two patients had simultaneous cytomegalovirus retinitis with other AIDS associated diseases (Kaposi's sarcoma in one patient and cytomegalovirus colitis in one patient). Treatment with ganciclovir (dihydroxy propoxymethyl guanine), a new antiviral medication related to acyclovir, resulted in regression of retinitis in all five patients treated with this drug. Our study shows that cytomegalovirus retinitis may be the initial manifestation of AIDS and that ganciclovir is a clinically effective antiviral agent for cytomegalovirus. PMID- 3030113 TI - Immunophenotypic characterization of the cutaneous exanthem of SIV-infected rhesus monkeys. Apposition of degenerative Langerhans cells and cytotoxic lymphocytes during the development of acquired immunodeficiency syndrome. AB - A T-cell tropic retrovirus, simian immunodeficiency virus (SIV), has recently been isolated from immunodeficient rhesus monkeys. This virus has remarkable similarities to human immunodeficiency virus (HIV), the etiologic agent of acquired immunodeficiency syndrome. Subsequent studies of simian infection with SIV have shown it to be a relevant animal model for studying the pathogenesis of AIDS in man. In both HIV-infected humans and SIV-infected monkeys, a cutaneous maculopapular eruption has been described. To date, the pathogenesis and possible relationship of these exanthema to the evolution of systemic immunosuppression have remained obscure. In this study, the mononuclear cell infiltrates that characterize skin rashes of SIV-infected rhesus monkeys were found to be composed predominantly of cells with phenotypic characteristics of cytotoxic/suppressor (T8+) lymphocytes and natural killer cells. Many of these cells expressed membrane-bound interleukin-2 receptor molecules. Double labeling and immunoelectron microscopy revealed these cells in direct contact with degenerative Langerhans cells within the epidermis and dermis. These observations suggest that the cutaneous rash associated with SIV infection may be the consequence of target cell injury of Langerhans cells by effector cells with cytotoxic potential. PMID- 3030114 TI - Adhesion of polymorphonuclear leukocytes to endothelium enhances the efficiency of detoxification of oxygen-free radicals. AB - Polymorphonuclear leukocytes can produce active oxygen species such as hydrogen peroxide and superoxide under various conditions. Because these substances can be toxic to cells, it is possible that the interaction between the circulating leukocytes and the blood vessel wall, either in normal circulation or during the acute inflammatory response, could damage the endothelial lining. Using an in vitro system of cultured endothelial cells and isolated polymorphonuclear leukocytes, we have measured the levels of detectable superoxide when neutrophils are attached to either endothelial monolayers or to plastic. Our results show that the levels of superoxide, on a per-cell basis, are lower when the neutrophils are attached to endothelium than when attached to plastic, even if the neutrophils are stimulated with phorbol myristate acetate. This is also reflected in data showing that no injury occurs to the endothelial cells, as measured by 51Cr release, under these same conditions. When endothelial cells are pretreated with an inhibitor of superoxide dismutase, diethyldithiocarbamate, the levels of superoxide detected are the same for neutrophils stimulated on plastic and those on the endothelial monolayer, suggesting that endothelial superoxide dismutase may remove a portion of the neutrophil-generated superoxide from the detection system. Further evidence for the role of endothelium in destroying superoxide is suggested by results that show that the level of detectable superoxide released from neutrophils attached to formalin-fixed endothelial monolayers is the same as that for neutrophils attached to plastic. It is important to note that with the inhibitor of superoxide dismutase present, the endothelial monolayers do not display enhanced 51Cr release under the conditions employed. When both endothelial catalase and glutathione reductase are inhibited, we detect increased 51Cr release from endothelial cells in response to stimulated neutrophils. Our results show that the endothelial cells are important in affecting the apparent reduction of toxic oxygen products derived from polymorphonuclear leukocytes attached to their surface. PMID- 3030115 TI - Mechanism of Escherichia coli alpha-hemolysin-induced injury to isolated renal tubular cells. AB - Alpha-hemolysin (AH) is a 110,000-dalton protein secreted extracellularly by certain Escherichia coli. This protein is an acknowledged virulence factor for E coli and recently has been implicated as an important determinant in the pathogenesis of E coli pyelonephritis. Recombinant engineered strains of E coli were used that varied only in their ability to secrete AH extracellularly. The effect of AH on vital dye exclusion, oxygen consumption rate (QO2) adenosine triphosphate (ATP) levels, superoxide (O2-) and hydrogen peroxide (H2O2) production in preparations of isolated rat cortical renal tubular cells (RTCs) was assessed. Approximately 5-10 pg of AH dramatically stimulated QO2 by nearly 150%. This was associated with a marked increase in production of O2- and H2O2, to 13.9 +/- 1.7 and 13.2 +/- 2.1 nM/mg cell protein, respectively (P less than 0.05), as well as a 38% decrease in cellular ATP. These biochemical effects were all seen after a 30-minute exposure to AH and by 120 minutes were associated with 15.7% +/- 1.1% of RTCs that were unable to exclude vital dye. The effect of AH on QO2 and O2- formation was prevented by pretreatment of RTCs with ouabain, which indicates that the effect of AH on oxygen metabolism is linked to Na-K ATPase activity. However, when ouabain-treated RTCs were exposed to AH, ATP remained depressed despite the inhibition of QO2 and O2- production. In contrast, in ouabain-pretreated RTCs, cell membrane integrity was dramatically protected, because only 2.4% +/- 0.4% of RTCs were not unable to exclude vital dye. Thus, the data demonstrate that E coli AH provokes at least two biochemical events that may be injurious to RTC: increased oxygen intermediates (O2- and H2O2 and ATP depletion. These findings with ouabain suggest that the first mechanism of injury may be a more proximate cause of cell death. Moreover, the data suggest that endogenous production of reactive oxygen molecules may be critical modulators of RTC membrane injury. PMID- 3030116 TI - Clonal T-cell populations in pityriasis lichenoides et varioliformis acuta (Mucha Habermann disease). AB - Patients with the skin disorder pityriasis lichenoides et varioliformis acuta (PLEVA) develop recurrent, self-healing papulonecrotic lesions that contain infiltrates of cytologically and antigenically normal T lymphocytes. DNA extracted from the lesions of 3 patients with PLEVA was analyzed for rearrangement of beta-T-cell receptor genes for the purpose of assessing the clonality of T lymphocytes within the tissues of this disease. Lesions from all 3 cases showed clonal gene rearrangements. In each of 2 cases from which two separate lesions were biopsied, identical rearrangements were found in specimens from both sites. DNA from a variety of inflammatory lesions obtained from patients with other types of skin diseases failed to show detectable rearrangements of beta-T-cell receptor genes. These results suggest that PLEVA represents a T-cell lymphoproliferative process, rather than an inflammatory disorder, as had been previously thought. PMID- 3030117 TI - Coxsackievirus B-3-induced myocarditis. Effect of sex steroids on viremia and infectivity of cardiocytes. AB - Male and female BALB/c mice were inoculated with various concentrations of coxsackievirus, group B, type 3 (CVB3), ranging from 10 to 10(7) plaque-forming units (PFU). Lower viral doses (greater than 10(2) PFU) induced severe myocarditis in male mice but caused little injury in females. With 10(7) PFU, females also developed severe disease. Females may be relatively resistant to CVB3-induced myocarditis because virus entry into the blood and heart is less effective. Males given 125I-CVB3 show approximately 2-4 and 20-fold more radioactivity in the peripheral blood and heart, respectively, than females. No differences were observed between the sexes in 125I-bovine serum albumin penetration. Sex steroid hormones influence viremia and virus localization; females given exogenous testosterone and progesterone demonstrate ten times more virus in their hearts than animals given estradiol. The hormones may act by increasing virus receptor expression on endothelial cells and myocytes. PMID- 3030118 TI - Vasoactive agents and production of thrombosis during intravascular coagulation. 3. Comparative effects of catecholamines. AB - Epinephrine (E), isoproterenol (I), and dopamine (D) were compared with norepinephrine (N) for production of microthrombi during thrombin-induced disseminated intravascular coagulation (DIC) in rabbits. Only catecholamines acting on alpha-adrenoreceptors produced glomerular capillary thrombosis (GCT) typical of the generalized Shwartzman reaction (GSR). Epinephrine produced GCT three times (P less than 0.05) less severe than that produced by N, but beta blockade with propranolol (P) rendered E equal to N in potency. I and D reduced fibrinogen consumption produced by thrombin. I (0.5-0.66 microgram/kg/min), as opposed to D, prevented the GSR produced by endotoxin in the pregnant rat and the cortisone-sensitized rabbit, and P increased the severity of the GSR in the pregnant rat. Alpha-adrenergic blockade with dibenzyline prevented the GSR produced by endotoxin in rats, whether pregnant, diabetic, or having a unilateral ureteral occlusion, and the classic reaction in rabbits, but not that produced in renal-hypertensive rats. Simultaneous alpha + beta stimulations by E triggered coronary and hepatic microthrombi, which were prevented by P. It is concluded that beta-adrenergic stimulation, as opposed to D-adrenergic stimulation, counterbalances alpha-adrenergic effects occurring in endotoxemia, which are required for production of the GSR in most models. These studies stress the risks and benefits of beta-blockade and provide additional evidence for the role of vasoactive agents and microcirculatory changes on selection of target organs for production of microthrombi during DIC. PMID- 3030119 TI - Pretranslational regulation of Na-K-ATPase in cultured canine kidney cells by low K+. AB - Long-term upregulation of the sodium pump [Na-K-adenosine triphosphatase (Na-K ATPase)] entails an increase in the number of enzyme molecules. We incubated Madin-Darby canine kidney (MDCK) cells in low K+ medium and studied the time course and magnitude of change in the relative abundance of the two Na-K-ATPase subunits (alpha and beta), in the synthesis rate of the subunits, and in the relative abundance of alpha- and beta-mRNA. When cells were incubated in medium containing 0.25 mM K+, intracellular Na+ increased from 25.2 +/- 0.9 (SE) mmol/l cell H2O to 69.8 +/- 9.6 at 4 h and 132 +/- 6 at 16 h. Cell K+ fell from 146 +/- 4 mmol/l cell H2O to 105 +/- 9 at 4 h and 42.3 +/- 4.7 at 16 h. The relative abundance of Na-K-ATPase subunits, measured with immunoblots of cell homogenates, increased such that after 24 h alpha was 1.71 +/- 0.33 and beta was 1.67 +/- 0.22 times control. After 8 h of K+ depletion, alpha-synthesis rate, measured by immunoprecipitation of pulse-labeled cells, increased to 2.30 +/- 0.50 and beta increased to 2.07 +/- 0.42 times control. The alpha- and beta-subunit mRNA abundance, measured by hybridizing alpha- and beta-cDNA probes to total RNA, increased within 30 min to 1.93 +/- 0.24 and 2.29 +/- 0.64 times control, respectively. We conclude that regulatory adjustments of Na-K-ATPase abundance involve an increase in translation after a rapid and coordinate increase in the concentrations of alpha- and beta-subunit mRNA. PMID- 3030120 TI - Swelling, NEM, and A23187 activate Cl(-)-dependent K+ transport in high-K+ sheep red cells. AB - In low K+ (LK) sheep red cells a significant fraction of the total ouabain resistant (OR) K+ flux is inhibited when Cl- is replaced by other anions of the Hofmeister series except Br- (Cl(-)-dependent K+ flux). In contrast, high K+ (HK) sheep red cells in isosmotic media did not possess any significant OR Cl(-) dependent K+ flux when Cl- was replaced by NO3- or I-. However, exposure to hyposmotic solutions, treatment with the sulfhydryl (SH) group reagent N ethylmaleimide (NEM) or with the bivalent metal ion (Me2+) ionophore A23187 in absence of external Me2+ caused a significant activation of Cl(-)-dependent K+ transport as measured with Rb+ as K+ congener. There was no Cl(-)-dependent Rb+ flux in A23187-treated cells when Mn2+, Mg2+, and Ca2+ were present at 1 mM concentrations, suggesting that cellular accumulation of these Me2+ is inhibitory. Similar to LK red cells, HK red cells failed to respond to A23187 when pretreated with NEM supporting the hypothesis proposed recently (Lauf, P. K. J. Membr. Biol. 88: 1-13, 1985) of a common mechanism of Cl(-)-dependent K+ transport activation. The magnitudes of the Cl(-)-dependent Rb+ fluxes in HK cells were much smaller than those elicited by identical treatments in LK red cells, and the effect of all interventions was not due to the presence of reticulocytes known to possess Cl(-)-dependent K+ transport.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3030121 TI - Energy metabolism of renal cell lines, A6 and MDCK: regulation by Na-K-ATPase. AB - The energy metabolism of two continuous cell lines of renal origin, MDCK (Madin Darby dog kidney) and A6 (toad kidney), was investigated by measuring the oxygen consumption (QO2) and lactate production (Jlac) by cells taken into suspension from monolayer cultures. Cells suspended from fully differentiated monolayers produce approximately 80% of their ATP requirements from oxidative metabolism. The interrelationship between ion transport and metabolism was determined by analyzing the ouabain sensitive components of intermediary metabolism under control conditions and after the stimulation of active Na-K transport with nystatin. In both cell lines, approximately 25% of the net rate of ATP production was inhibited by ouabain. Ouabain inhibited Jlac by 40% in MDCK and 45% in A6 cells, whereas QO2 was decreased by only 20% in both cell lines. In the presence of 0.05 mg nystatin/mg cell protein, ouabain sensitive Jlac increased by 75% in MDCK and was more than doubled in A6, whereas the ouabain-sensitive QO2 was not statistically different than control. This preferential stimulation of glycolysis with nystatin was not due to a limited capacity of mitochondrial oxidative phosphorylation since nystatin treatment of cells incubated without glucose (no glycolysis) significantly elevated the rate of QO2. These data demonstrate that aerobic glycolysis is more sensitive than is QO2 to changes in hydrolytic activity of the Na-K-adenosine triphosphatase (ATPase), in both cell lines. PMID- 3030122 TI - Isolation, growth, and characterization of a gluconeogenic strain of renal cells. AB - LLC-PK1 cells, derived from pig kidney, retain several properties of the proximal tubule, but are incapable of gluconeogenesis, due to the lack of fructose-1,6 bisphosphatase (FBPase) [Am. J. Physiol. 248 (Cell Physiol. 17): C181-185, 1985]. Cells incapable of gluconeogenesis require a hexose, pentose, or nucleoside to provide ribose-5-phosphate for RNA biosynthesis. To induce or select cells that express FBPase activity, we cultured LLC-PK1 cells in glucose-free medium. We obtained cells (designated LLC-PK1-FBPase+) that express FBPase activity and are capable of growing in the complete absence of sugars or nucleosides. The cells have apical membrane enzyme activities that differ from those of wildtype cells. Tests of metabolic flow through the gluconeogenic pathway, using 3 mercaptopicolinic acid, a specific inhibitor of phosphoenolpyruvate carboxykinase, confirmed that the cells are gluconeogenic. LLC-PK1-FBPase+ cells grown in medium containing 5 mM glucose for five weekly passages continued to express FBPase activity and apical membrane enzyme activities characteristic of the FBPase+ strain. When switched back to glucose-free medium, they proliferated well. The strain appears to be stable. It should provide a model for studying the relationship between gluconeogenesis and other proximal tubule functions. An incidental finding is that in both strains, the activity of lactate dehydrogenase varied directly with the concentration of glucose in the growth medium, indicating that the expression of lactate dehydrogenase may be regulated by glucose or a metabolite of glucose. PMID- 3030123 TI - WR-2721 inhibits parathyroid adenylate cyclase. AB - WR-2721 [S-2-(3-aminopropylamino)ethylphosphorothioic acid] is a chemoprotective and radioprotective agent that has been shown to lower serum calcium in dogs and in humans. This is secondary both to impaired release of Ca2+ from bone and diminished secretion of parathyroid hormone (PTH) from parathyroid glands. Because cAMP plays a role in the regulation of PTH secretion and WR-2721 has been shown to lower cAMP levels in radiated mouse spleen, we investigated the effects of WR-2721 on cAMP production in dispersed bovine parathyroid cells. Additionally, we studied the adenylate cyclase in plasma membranes from normal bovine parathyroid glands after exposure to WR-2721. With parathyroid cells incubated at 0.5 mM Ca2+, addition of WR-2721 in concentrations ranging from 0.02 to 2.0 mM resulted in a progressive decrease in intracellular cAMP (42-50%, respectively). In plasma membranes of bovine parathyroid cells a dose-dependent decrease in adenylate cyclase activity was noted. Inhibition of the cyclase was seen over a wide range of Mg2+ concentrations (2.5-40 mM). WR-2721 inhibited both basal and NaF, Gpp(NH)p, forskolin, and pertussin toxin-stimulated adenylate cyclase. These data suggest that WR-2721 inhibits the activity of parathyroid adenylate cyclase. PMID- 3030124 TI - Mineralocorticoid specificity of renal type I receptors: in vivo binding studies. AB - We have injected rats with [3H]aldosterone or [3H]corticosterone, plus 100-fold excess of the highly specific glucocorticoid RU 28362, with or without excess unlabeled aldosterone or corticosterone and compared type I receptor occupancy in kidney and hippocampus. Thirty minutes after subcutaneous injection [3H]aldosterone was well retained in renal papilla-inner medulla, renal cortex outer medulla, and hippocampus; in contrast, [3H]corticosterone was well retained only in hippocampus. Competition studies for [3H]aldosterone binding sites showed corticosterone to be a poor competitor in the kidney compared with hippocampus. Time-course studies, with rats killed 10-180 min after tracer administration, showed very low uptake/retention of [3H]corticosterone by kidney; in hippocampus [3H]corticosterone retention was similar to that of [3H]aldosterone in kidney, and retention of [3H]aldosterone by hippocampus was much more prolonged than of either tracer in any other tissue. Studies in 10-day-old rats, with very low levels of corticosteroid binding globulin (CBG), showed a high degree of aldosterone selectivity in both zones of the kidney, whereas [3H]aldosterone and [3H]corticosterone were equivalently bound in hippocampus. We interpret these data as evidence for a mechanism unrelated to extravascular CBG conferring mineralocorticoid specificity on renal type I receptors and propose two models derived from our findings consistent with such differential selectivity. PMID- 3030125 TI - Polymyxin B selectively inhibits insulin effects on transport in isolated muscle. AB - Polymyxin B (PMB), a cyclic decapeptide antibiotic, inhibits the hypoglycemic effect of insulin in vivo. To elucidate the mechanism of PMB action, we have studied its effect in vitro on insulin-stimulated pathways in the mouse skeletal muscle. PMB, added to the incubation mixture, specifically inhibited insulin stimulated 2-deoxyglucose transport and alpha-aminoisobutyric acid uptake in the isolated soleus muscle but did not affect the basal rates of transport (measured in the absence of insulin). PMB did not alter insulin binding and hexokinase activity. PMB effect was observed at all deoxyglucose concentrations tested, and PMB was also able to inhibit vanadate-stimulated glucose transport. By contrast, insulin activation of glycogen synthase was not prevented by PMB. Basal and maximally insulin-stimulated insulin receptor tyrosine kinase activity, tested in a cell-free system, was similar for both autophosphorylation and phosphorylation of exogenous substrates in the absence or in the presence of PMB. Furthermore, the insulin sensitivity of the kinase was increased in the presence of PMB. Our results suggest that the anti-insulin effect of PMB observed in vivo is due to an inhibition of insulin-stimulated glucose transport in the skeletal muscle perhaps through a specific blockade of the insulin-induced translocation of the glucose carriers. PMID- 3030126 TI - Formation and actions of calcium-mobilizing messenger, inositol 1,4,5 trisphosphate. AB - A variety of surface membrane receptors can activate a phospholipase C, which degrades phosphatidylinositol 4,5-bisphosphate liberating a calcium mobilizing second messenger, inositol 1,4,5-trisphosphate [(1,4,5)IP3]. The coupling of surface receptors to the phospholipase C involves one or more guanine nucleotide dependent regulatory proteins that are similar but not identical to those that regulate adenylate cyclase. (1,4,5)IP3 has been shown to release Ca2+ from a portion of the endoplasmic reticulum and is believed responsible for the initial phase of Ca2+ mobilization ascribed to internal Ca2+ release. (1,4,5)IP3 acts by binding to a specific receptor that either is a component of, or regulates, a Ca2+ ion channel. The release of Ca2+ from the (1,4,5)IP3-sensitive component of the endoplasmic reticulum may secondarily activate the second phase of Ca2+ mobilization, which involves Ca2+ entry. (1,4,5)IP3 is metabolized by two pathways. One involves the action of a 5-phosphatase that degrades (1,4,5)IP3 to inositol 1,4-bisphosphate, whereas the other involves a 3-kinase that phosphorylates (1,4,5)IP3 to produce inositol 1,3,4,5-tetrakisphosphate. The significance of this dual metabolism is not known, but it may be important in rapidly extinguishing the Ca2+-releasing activity (1,4,5)IP3. PMID- 3030127 TI - Effects of ethanol on gastric epithelial cell phospholipid dynamics and cellular function. AB - The lipid profile of isolated gastric superficial epithelial cells (SEC) was evaluated by proton nuclear magnetic resonance spectroscopy (1H-NMR). The most conspicuous resonance band in SEC spectra was due to the protons of +N(CH3)3 groups of phosphatidylcholine and, to a lesser degree, other phospholipid derivatives, on the basis of their chemical shift and addition of purified phospholipids. NMR of cell lysates and phospholipid extracts of SEC in deutero chloroform provided further spectral resolution of these components. Phospholipase or ethanol treatments of SEC produced membrane disorganization reflected as increased peak intensity of the phospholipid signals. In addition, ethanol, in a dose-dependent manner, attenuated paranitrophenyl phosphatase activity, which correlated with inhibition of total and ouabain-sensitive 86Rubidium chloride uptake by SEC. This study suggests that NMR used in conjunction with other biochemical techniques can monitor SEC membrane structure function relationships. NMR is a potentially powerful noninvasive probe to show changes in lipid membrane organization induced by low concentrations of ethanol (1%) and may indicate an early sign of "cytotoxicity" in intact SEC. PMID- 3030128 TI - Characteristics of glycyl-L-proline transport in intestinal brush-border membrane vesicles. AB - A proton-peptide symport mechanism has been postulated for transport of dipeptides in rabbit intestinal and renal brush-border membrane vesicles (BBMV). We have investigated the effects of a transmembrane potential (in mouse) and an inwardly directed proton gradient (in mouse, rabbit, and human) on the transport of glycyl-L-proline in intestinal BBMV. Membrane potential alterations, induced by permeant anions or generated by a K+-diffusion potential in the presence of valinomycin, did not accelerate the uptake of glycyl-L-proline. In contrast, in parallel experiments the uptake of D-glucose, whose cotransport system is electrogenic, was markedly increased by an interior negative membrane potential. Thus the transport of glycyl-L-proline in mouse intestinal BBMV is not electrogenic. Further studies on the effect of a proton gradient (extravesicular pH 5.5; intravesicular pH 7.5) on transport of glycyl-L-proline revealed an absence of stimulation of glycyl-L-proline transport and lower uptake rates in the presence of a proton gradient. The simultaneous presence of an interior negative membrane potential and an inwardly directed proton gradient did not accelerate the transport of glycyl-L-proline. These results demonstrate that the transport of glycyl-L-proline in mouse intestinal BBMV is neither electrogenic nor energized by an inwardly directed proton gradient. Likewise, pH gradients do not stimulate glycyl-L-proline uptake in either rabbit or human BBMV. PMID- 3030129 TI - Mechanism of NaCl transport-stimulated prostaglandin formation in MDCK cells. AB - Recently we have found that stimulation of NaCl transport in high-resistance MDCK cells enhances their prostaglandin formation. In the present study, we investigated the mechanisms by which prostaglandin formation could be linked to the ion transport in these cells. We found that stimulation of transport caused a transient stimulation of prostaglandin formation lasting 5-10 min. The rise in prostaglandin formation was paralleled by a rise of free intracellular arachidonic acid. Analysis of membrane lipids revealed that the rise of free arachidonic acid was paralleled by a loss of arachidonic acid from polyphosphoinositides. We failed to obtain indications for the stimulation of calcium-dependent phospholipase A2. However, we did obtain evidence that the incorporation of arachidonic acid into phospholipids was diminished during stimulation of ion transport, indicating a decreased rate of reesterification. Despite the fact that there was no significant fall in total cellular ATP on stimulation of ion transport, we found a high and transient rise of lactate production of the cells on stimulation of the ion transport indicating an alteration of the ADP/ATP ratio. Moreover, prostaglandin formation and lactate formation were linearly correlated in this situation. When glucose utilization was inhibited by mannoheptulose, the rise in lactate formation was abolished, whereas that of PG formation was unaltered, indicating that lactate formation and prostaglandin formation were not causally linked on stimulation of ion transport. Our results suggest that an increase in the rate of sodium chloride transport by MDCK cells stimulates formation by an inhibition of reesterification of free arachidonic acid.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3030130 TI - Calcium-mediated cyclic AMP inhibition of Na-H exchange in small intestine. AB - 8-Bromo cyclic AMP (cAMP) (10(-4) M) inhibits Na absorption in isolated chicken enterocytes as has been reported previously. Direct measurements of intracellular pH (pHi) using 5,6-carboxyfluorescein diacetate showed that both 8-bromo cAMP and the diuretic amiloride (10(-3) M) stimulated a persistent decrease in pHi of approximately 0.1 pH units, effects that were Na dependent and were not additive when cells were stimulated with both agents. These results suggest inhibition of an amiloride-sensitive Na/H exchange by cAMP. Direct measurements of intracellular Ca [Ca]i were also made using quin 2. 8-Bromo cAMP (10(-4) M) stimulated an immediate and persistent (greater than 10 min) increase in [Ca]i of approximately 20 nM, an effect that was not dependent on extracellular Ca. Pretreatment of cells with the specific calmodulin inhibitor calmidazolium (10( 7) M) and the intracellular Ca-buffering agent MAPTAM blocked cAMP's effects on pH and Na uptake, but did not interfere with amiloride's effects. An increase in [Ca]i stimulated by the Ca ionophore A23187 (10(-6) M) was sufficient by itself to decrease pHi and inhibit amiloride-sensitive Na influx in isolated enterocytes. We conclude that cAMP stimulates the release of endogenous Ca in isolated enterocytes. This increase in [Ca]i appears to be essential for inhibition of amiloride-sensitive Na-H exchange by this cyclic nucleotide. PMID- 3030131 TI - Compartmentation of carbohydrate metabolism in vascular smooth muscle. AB - In vascular smooth muscle, oxidative phosphorylation and glycolysis are independently regulated. Previous studies indicated that the independent regulation of these pathways was related to a compartmentation of carbohydrate metabolism. To further study carbohydrate metabolism, glucose transport and the incorporation of radiolabel from glucose into glycogen and lactate were measured after the oxidative and glycolytic pathways were independently altered. Ouabain stimulated mechanical activity, oxygen consumption, and glycogenolysis, whereas lactate production was decreased. Although glycogenolysis was substantial, glucose was the only substrate for lactate, indicating that intermediates derived from glycogen do not mix with those from glucose uptake. Thus glycogenolysis and glycolysis are carried out by independent enzymatic pathways. Insulin-stimulated lactate production and glucose transport without affecting the other parameters. Again, lactate was produced only from glucose. Phenytoin decreased isometric tension and oxygen consumption, whereas stimulating lactate production and glycogenolysis. Glycogen was the primary substrate for the lactate produced. Our findings indicate that the compartmentation of substrate utilization is ascribable to the coordination of glycogenolysis with increases in oxygen consumption and the coupling of glycolysis to the Na-K-adenosine triphosphatase. The coupling of independent energy providing pathways to specific endergonic processes indicates a mechanism by which cellular energetic efficiency may be optimized. PMID- 3030132 TI - Effect of N6-(L-2-phenylisopropyl)adenosine and insulin on cAMP metabolism in 3T3 L1 adipocytes. AB - The antilipolytic effect of N6-(L-2-phenylisopropyl)-adenosine (PIA), an adenosine analogue thought to act via cell surface receptors, was investigated in 3T3-L1 adipocytes. PIA (1 microM) was as effective as 1 nM insulin in reducing lipolysis stimulated by 1 nM isoproterenol and more effective than insulin at higher isoproterenol concentrations. In intact adipocytes, PIA reduced isoproterenol-induced cyclic AMP (cAMP) accumulation and increased particulate cAMP phosphodiesterase. In particulate preparations PIA suppressed isoproterenol stimulation of adenylate cyclase. PIA was more effective than 5'-N ethylcarboxamide adenosine (NECA) or adenosine in inhibiting adenylate cyclase and activating phosphodiesterase. In intact adipocytes, two agents with so-called "insulin-like" activities, i.e., anti-insulin receptor antibodies and wheat germ agglutinin (WGA), also increased particulate cAMP phosphodiesterase. Pertussis toxin, which inhibits stimulation of the particulate cAMP phosphodiesterase by insulin (but not by isoproterenol), also inhibited the effects of PIA, anti insulin receptor antibodies, and WGA. In 3T3-L1 cells, PIA appears to inhibit lipolysis by inhibiting adenylate cyclase and stimulating phosphodiesterase; these effects of PIA, as well as those of anti-insulin receptor antibodies and WGA on phosphodiesterase, may be mediated via guanyl nucleotide-binding proteins. PMID- 3030133 TI - Metabolic basis of diethylaminoethoxyhexestrol-induced phospholipid fatty liver. AB - Diethylaminoethoxyhexestrol caused a foam cell lipidosis in humans characterized by phospholipid storage in the liver, spleen, and other tissues, and this represents the first description of acquired lipidosis caused by a drug. It has been proposed that diethylaminoethoxyhexestrol causes phospholipid fatty liver by concentrating in lysosomes and inhibiting phospholipases but it has not previously been possible to measure the intralysosomal concentration of diethylaminoethoxyhexestrol. In this paper we report for the first time the intralysosomal concentration of this drug in rats. After a single oral dose of diethylaminoethoxyhexestrol (100 mg/kg) the intralysosomal concentration was 7.9 mM at 2.5 h, 15.6 mM at 12 h, and 20.9 mM at 24 h, respectively. The total phospholipid content of lysosomes in drug-treated rats increased 1.9-, 6.0-, and 7.6-fold over control at 2.5, 12, and 24 h, respectively. Purified lysosomal phospholipase A1 was strongly inhibited by diethylaminoethoxyhexestrol in vitro. In phospholipid fatty liver, phospholipid accumulation in lysosomes appears to be caused by the presence of diethylaminoethoxyhexestrol in lysosomes at concentrations estimated to be 7.9-20 mM, because drug levels above 1 mM completely block the activity of purified lysosomal phospholipase A1 in vitro. PMID- 3030134 TI - Epinephrine potentiates adenosine-stimulating effect on glucagon secretion. AB - The aim of the present work is to investigate a possible interaction on glucagon secretion between adenosine, a compound released by tissues in energy-deficient states, and epinephrine, the hormone of stress largely implicated in such conditions. The study was performed using the isolated perfused rat pancreas in presence of a physiological glucose concentration (5 mM). Epinephrine administered at a low concentration (0.01 microM) was ineffective on glucagon secretion, and adenosine at 1.65 microM was previously shown to be moderately stimulating. This nucleoside alone induced a transient increase of glucagon secretion rate that peaked at 300% of basal value at 2 min; in presence of epinephrine (ineffective per se) the rise induced by the nucleoside alone was doubled. This potentiating effect was not observed with the neurotransmitter norepinephrine at the dose tested. Propranolol (1 microM) did not alter the potentiating effect of epinephrine but this effect was completely suppressed by the alpha-blocker, phenoxybenzamine (6 microM). In conclusion epinephrine potentiates an adenosine-stimulating effect on glucagon secretion; this effect seems more specific for the adrenal medulla hormone epinephrine, since norepinephrine at the same dose is ineffective; it is mediated via alpha adrenergic receptors. It is attractive to speculate that epinephrine and adenosine act in potentiating synergism on glucagon secretion; this might be of physiological importance during stressful energy-deficient situations. PMID- 3030135 TI - Site and mechanisms of action of kinins in rat ileal mucosa. AB - Kinin-induced secretion in the intestine is accompanied by marked increases in mucosal adenosine 3', 5'-cyclic monophosphate (cAMP) and prostanoids that undoubtedly contribute to the overall secretory response. The cellular site at which these effects are initiated is unclear although a recent hypothesis has suggested the epithelial cell as the target for kinin action (6). We have investigated the effects of kallidin on the phospholipase-prostanoid-cAMP pathway in whole ileal mucosa and in epithelial cells isolated from the same tissue in the rat. Kallidin (1 microM) stimulated a marked rise (24 to 30-fold) in PG (prostaglandin) E2 release from the serosal surface of stripped ileal mucosa within 1-2 min, which correlated closely with the rise in mucosal short-circuit current. Mucosal cAMP levels were also increased two to threefold by kallidin. However, kinins were unable to elicit effects under the same conditions in suspensions of viable epithelial cells. PGE2 release was unaffected by kallidin or bradykinin at concentrations up to 100 microM, whereas cAMP levels could be stimulated by forskolin and PGE2 but not by kinin. Studies of intestinal phospholipase A2 (PLA2) activity also suggest a nonepithelial site for kinin action. This enzyme is responsible for liberating arachidonic acid from cellular phospholipids and has been shown to be activated by kinins in several tissues. In the intestine, PLA2 activity was found to be concentrated within the subepithelium with significantly lower levels in the epithelium itself. In addition, kallidin was unable to influence phospholipid labeling (an indirect measure of PLA2 activity) in cells incubated with [14C]arachidonic acid.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3030136 TI - The kidney in potassium depletion. I. Na+-K+-ATPase activity and [3H]ouabain binding in MCT. AB - The effect of potassium depletion on renal Na+-K+-ATPase was studied in rats. K depletion produced a striking, time-dependent increase in Na+-K+-ATPase activity of the outer medullary collecting tubules (inner stripe; MCTis). After 3 wk on the K-free diet, when the urine was almost potassium-free, Na+-K+-ATPase activity in MCTis was over fourfold higher than in control animals (2,964 +/- 185 vs. 645 +/- 108 pmol X mm-1 X h-1). Repletion of potassium restored enzyme activity to base line within 7 days (t1/2 = 3.8 days), which corresponds to the catabolic rate of the renal enzyme, suggesting the cessation of enhanced synthesis that took place during K deprivation. Changes in Na+-K+-ATPase activity and aldosterone levels during both K depletion and repletion occurred in opposite directions and were therefore independent of each other. [3H]Ouabain binding to intact MCTis, reflecting the number of pump sites on the basolateral membrane, was similar in K-depleted and control animals; in contrast, tubule permeabilization that exposes additional pump units to the ligand, unmasked a nearly fourfold increase in [3H]ouabain binding (50.0 +/- 6.8 vs. 13.2 +/- 1.7 fmols X mm-1) in K-depleted rats, comparable to the increment in Na+-K+-ATPase activity. These results show that K depletion leads to a marked increase in Na+ K+-ATPase activity of MCTis, and suggest that the new enzyme units are located at a ouabain-inaccessible site in the intact tubule, i.e., either in an intracellular compartment or at the luminal membrane, where they may be involved in potassium reabsorption. PMID- 3030137 TI - The kidney in potassium depletion. II. K+ handling by the isolated perfused rat kidney. AB - In a companion paper we reported a large increment in Na+-K+-ATPase activity and [3H]ouabain binding in the inner stripe of outer medullary collecting tubules from K-depleted rats. To test the hypothesis that the increased number of Na+-K+ pumps in these animals may be involved in potassium reabsorption we examined the effect of ouabain on K excretion by isolated, perfused kidneys from rats fed a K free diet for 3 wk. Kidneys from K-depleted rats retain potassium avidly, both the fractional (FEK) and absolute K excretion being approximately fivefold lower than in control kidneys. Ouabain (5 mM) increased FEK in kidneys from each K depleted rat [mean from 8.5 to 34.4%, P less than 0.001, at average (4.5 mM) perfusate K]; similar results were obtained when kidneys were perfused with low (approximately 2 mM) and high (greater than or equal to 8 mM) potassium concentrations. In contrast, ouabain produced a variable effect in control kidneys, that depended on the perfusate potassium concentration. In K-depleted rats amiloride (10(-4) M) did not significantly alter K excretion and did not block the ouabain-induced kaliuresis, suggesting that the latter is not due to enhanced secretion secondary to increased distal fluid delivery. These results provide evidence for ouabain-sensitive potassium reabsorption in kidneys of chronically K-depleted rats, and suggest an explanation for the increased Na+-K+ ATPase observed in such animals. PMID- 3030138 TI - N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline, amiloride analogues, and renal Na+/H+ antiporter. AB - N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) is a carboxyl-activating agent and has been shown to inhibit the renal Na+/H+ antiporter. The purposes of the present studies were to characterize the kinetics of inhibition of the Na+/H+ antiporter by EEDQ and to determine whether amiloride analogues affect the ability of EEDQ to inhibit the rate of Na+/H+ exchange. Brush-border membrane vesicles (BBMV) were prepared from rabbit kidneys; Na+/H+ exchange rate was assessed by the fluorescence quenching of acridine orange. EEDQ produced a concentration-dependent inhibition of Na+/H+ exchange; the effect was to decrease the maximum activity (Vmax) from 5.51 to 2.03 fluorescence units X mg protein-1 X S-1) and Km (from 14.1 to 8.7 mM) compared with control BBMV. Pretreatment of BBMV with amiloride before the addition of EEDQ maintained both Vmax and Km at values that were not significantly different from those for control BBMV. Compared with a series of analogues, amiloride was only the third most potent inhibitor of the rabbit renal Na+/H+ antiporter; amiloride, however, provided the greatest protection against inhibition of the antiporter by the subsequent addition of EEDQ. These findings suggest that the 2-carbonylguanidininum moiety and 6-chloro atom are important for binding of amiloride to sites at or near the antiporter; the group at position 5 is important in determining the ability of amiloride to protect against inhibition of the Na+/H+ antiporter by EEDQ. Finally, the ability of amiloride to protect against inactivation of the renal Na+/H+ antiporter by EEDQ is reversible. PMID- 3030139 TI - Effects of thyroid hormone on beta-adrenergic responsiveness of aging cardiovascular systems. AB - We have compared the effects of beta-adrenergic stimulation on the heart and peripheral vasculature of young (2-mo-old) and older (12-mo-old) rats both in the presence and absence of triiodothyronine (T3)-induced hyperthyroidism. The hemodynamic consequences of T3 treatment were less prominent in the aged hyperthyroid rats compared with young hyperthyroid rats (both in intact and pithed rats). There was a decrease in sensitivity of chronotropic responsiveness to isoproterenol in older pithed rats, which was apparently reversed by T3 treatment. The number and affinity of myocardial beta-adrenergic receptor sites measured by [125I]cyanopindolol were not significantly different in young and older control rats; also, beta-receptor density increased to a similar extent in both young and older T3-treated rats. The ability of isoproterenol to relax mesenteric arterial rings, markedly blunted in older rats, was partially restored by T3 treatment without their being any change in isoproterenol-mediated relaxation in the arterial preparations from young rats. The number and affinity of the beta-adrenergic receptors measured in the mesenteric arteries was unaffected by either aging or T3 treatment. The data suggest that effects of thyroid hormone and age-related alterations of cardiovascular responsiveness to beta-adrenergic stimulation are interrelated in a complex fashion with a net result that the hyperkinetic cardiovascular manifestations in hyperthyroidism are attenuated in the older animals. PMID- 3030140 TI - Alterations in cardiac sarcolemmal Ca2+ pump activity during diabetes mellitus. AB - Diabetes mellitus is frequently associated with a primary cardiomyopathy. The mechanisms responsible for this heart disease are not clear, but an alteration in myocardial Ca2+ transport is believed to be involved in its development. Even though sarcolemma plays a crucial role in cellular Ca2+ transport, little appears to be known about its Ca2+ transporting capability in the diabetic myocardium. In this regard, we have examined the status of the cardiac sarcolemmal Ca2+ pump during diabetes mellitus. Purified sarcolemmal membranes were isolated from male Wistar diabetic rat hearts 8 wk after streptozotocin injection (55 mg/kg iv). Ca2+ pump activity assessed by measuring its Ca2+-stimulated adenosine triphosphatase and Ca2+-uptake ability in the absence and presence of calmodulin was significantly depressed in the diabetic myocardium relative to controls. These results did not appear to have been influenced by the minimal sarcoplasmic reticular and mitochondrial contamination of this membrane preparation. Hence, it appears that the sarcolemmal Ca2+ pump is defective in the diabetic myocardium and may be involved in the altered Ca2+ transport of the heart during diabetes mellitus. PMID- 3030142 TI - Ammonium ion substitutes for K+ in ATP-dependent Na+ transport by basolateral membrane vesicles. AB - Ion-transporting cells from posterior gills of blue crabs (Callinectes sapidus) acclimated to low salinity were used as starting material for the preparation of microsomal membrane vesicles by density gradient centrifugation. The Na+-K+ adenosinetriphosphatase (ATPase)-enriched basolateral vesicles were loaded with KCl- or NH+4-containing medium by dilution and centrifugation, and initial rates of 22Na+ uptake into the vesicles were measured by a rapid filtration procedure. Varying the extravesicular sucrose concentration altered equilibrium uptake of 22Na+, indicating the existence of osmotically sensitive vesicles. Monensin, a sodium-specific ionophore, enhanced passive uptake of 22Na+ across the vesicle membrane in the absence of ATP. With 100 mM KCl in the intravesicular medium, addition of ATP to the extravesicular medium increased initial rates of 22Na+ uptake 10- to 20-fold over levels measured without ATP. A nonhydrolyzable ATP analog failed to stimulate 22Na+ uptake. Intravesicular K+ could be replaced by NH+4 but not by choline. With NH+4 as counterion, Na+ transport was inhibited by digitoxin, but valinomycin had no effect. A study of the kinetic effects of intravesicular K+ and NH+4 on initial rates of 22Na+ uptake indicated the existence of two classes of binding sites, one responding to counterion concentrations in the millimolar range and a second class responding to counterion concentrations over 50 mM. Our results indicate that ATP-dependent 22Na+ uptake by membrane vesicles from Callinectes sapidus gill, mediated by Na+ K+-ATPase, can utilize either K+ or NH+4 as counterion. PMID- 3030141 TI - Ibuprofen prevents thrombin-induced lung vascular injury: mechanism of effect. AB - We compared pulmonary vascular responses with thrombin-induced pulmonary microembolism in awake sheep pretreated with ibuprofen or meclofenamate. Control sheep with lung lymph fistulas (n = 6) were challenged with 80 U/kg of alpha thrombin, the native enzyme. Two groups of similarly prepared sheep received the cyclooxygenase inhibitors, ibuprofen (n = 4) or meclofenamate (n = 4), before the thrombin challenge. Thrombin-induced pulmonary microembolism in control sheep increased pulmonary lymph flow and lymph protein clearance (lymph flow X lymph-to plasma protein concentration ratio). These lymph changes were associated with sustained increases in mean pulmonary arterial pressure (Ppa) and pulmonary vascular resistance (PVR) and a decrease in circulating neutrophil count. The steady-state pulmonary hemodynamic and lymph responses to thrombin persisted after cyclooxygenase inhibition with meclofenamate, although the increases in pulmonary lymph flow and lymph protein clearance were delayed. In contrast, ibuprofen reduced pulmonary lymph flow, lymph protein clearance, Ppa, and PVR responses. Neutrophil count in control and meclofenamate groups remained below base line after thrombin, whereas neutrophil count returned to base line in the ibuprofen group within 90 min after thrombin. Ibuprofen resulted in a greater decrease in vitro neutrophil adherence to pulmonary endothelium induced by serum generated by clotting whole blood with alpha-thrombin as compared with meclofenamate. Results indicate that ibuprofen attenuates the increases in Ppa and PVR and markedly attenuates the increase in pulmonary transvascular protein clearance after thrombin. The protective effect of ibuprofen may be due to reduction of neutrophil sequestration in lung. PMID- 3030143 TI - Angiotensin II does not alter ACTH responses to hypoglycemia in conscious dogs. AB - These experiments were designed to test for an interaction between angiotensin II (ANG II) and stress in the control of plasma adrenocorticotropin hormone (ACTH), corticosteroids, and aldosterone. The stimulus to ACTH used in this study was insulin-induced hypoglycemia, a stimulus that does not increase plasma ANG II concentrations. Five trained dogs with exteriorized carotid arteries were studied. Each dog was infused with ANG II intravenously (10 ng X kg-1 X min-1) or into the carotid artery (1 ng X kg-1 X min-1) or with saline (iv) for 80 min. Twenty minutes after the start of the infusion, insulin (0.10 U/kg iv) was injected. Intravenous infusion of ANG II increased mean arterial pressure (MAP) and plasma aldosterone concentrations but did not increase ACTH or corticosteroid responses to hypoglycemia. Intracarotid infusion of ANG II did not increase MAP and also failed to increase ACTH and corticosteroid responses to hypoglycemia. Since ANG II infusions did not increase basal corticosteroids, the failure of ANG II to stimulate ACTH is not a result of steroid negative feedback. Thus it appears that increased plasma ANG II concentrations do not increase ACTH responses to hypoglycemic stress. PMID- 3030144 TI - Basolateral glucose transport by intestine of teleost, Oreochromis mossambicus. AB - Transport characteristics of D-glucose by isolated basolateral membrane vesicles of the teleost fish, Oreochromis mossambicus, were measured. Specific activity of the vesicle Na-K-adenosinetriphosphatase was increased 11-fold, whereas specific activities of brush-border and organelle membrane enzymes were enriched only 0.3- to 0.8-fold. Vesicles had diameters of 0.1-0.4 micron, 70% of vesicles were leaky (unsealed), and 60% of sealed vesicles were inside out. D-Glucose transport occurred by stereospecific facilitated diffusion, independent of 120 mM gradients of either NaCl or KCl, and was inhibited by sulfhydryl reagents, phloretin, and cytochalasin B, but not by phloridzin. Competition studies with a range of sugars demonstrated that aldohexoses in the C-1 chair conformation were preferred substrates and probably share the same carrier. Kinetic analysis of glucose influx yielded a Kt of 10 mM and a Jmax of 3,910 pmol X mg protein-1 X min-1. Fish intestinal basolateral D-glucose transport closely resembles that of mammalian or avian intestinal epithelia and of red blood cell plasma membrane. The magnitude of transport is much lower in fish than in other vertebrates, which may be related to lower metabolic rates in these poikilotherms. PMID- 3030145 TI - Mandatory outpatient treatment. PMID- 3030147 TI - Kupffer cells in hepatocellular adenomas. AB - Hepatocellular adenomas are usually visualized as defects on technetium-99m sulfur colloid liver scans, a fact which has been attributed to the absence of phagocytic Kupffer cells in the tumors. To determine whether this is true, seven hepatocellular adenomas were subjected to immunoperoxidase staining for lysozyme, a marker of mononuclear phagocytes. The Kupffer cells were counted in the tumors and surrounding non-neoplastic liver. All hepatocellular adenomas studied were found to contain Kupffer cells. Three tumors had fewer Kupffer cells than the surrounding liver. Three had about the same number as the surrounding liver, and one had more Kupffer cells than the non-neoplastic liver. Thus, the lack of phagocytosis of colloid in liver scans is probably due to something other than a deficiency of Kupffer cells in the hepatocellular adenomas. PMID- 3030146 TI - Risk factors for human immunodeficiency virus (HIV) infections in homosexual men. AB - To clarify risk factors for infection with the human immunodeficiency virus (HIV) we selected at random 785 homosexual men who had participated in studies of hepatitis B in San Francisco in 1978-80 for a follow-up study of the acquired immunodeficiency syndrome. Although most had not been contacted in over five years, 492 (63 per cent) were located and enrolled. The 240 (67 per cent) who had developed antibodies to HIV, as measured by an enzyme-linked immunosorbent assay (ELISA), were compared with 119 who had remained seronegative. In multivariate analyses, receptive anal intercourse with ejaculation by nonsteady sexual partners, many sexual partners per month, and other indicators of high levels of sexual activity were highly associated with seroconversions. None of the sexual practices that we studied appeared to offer protection against HIV infection. PMID- 3030148 TI - Cystic epithelial-stromal tumor of the seminal vesicle. AB - A cystic tumor composed of atypical glands in a cellular stroma arose in the pelvis of a 49-year-old man. Two years later an identical tumor was again excised from the pelvis. Morphologic, immunohistochemical and ultrastructural studies indicate that this neoplasm arose in the seminal vesicle, possibly from a seminal vesicle cyst. The tumor did not involve the prostate gland, and immunohistochemical stains for prostate-specific antigen and prostatic acid phosphatase were negative. Ultrastructural study showed that both the glandular and mesenchymal components of the tumor recapitulated features of normal seminal vesicle, further establishing origin from this site. This tumor resembles the rare cystadenoma of the seminal vesicle, yet the cytologic atypia suggests low grade malignant potential. Following the second excision, the patient has had a disease-free interval of 18 months. Long term follow-up and recognition of additional cases is necessary to define the biologic potential of this unusual tumor. PMID- 3030149 TI - Adenomyoepithelial adenosis. PMID- 3030150 TI - Studies of the coagulation system in arenaviral hemorrhagic fever: experimental infection of strain 13 guinea pigs with Pichinde virus. AB - Significant coagulation abnormalities were associated with experimental infection of strain 13 guinea pigs with Pichinde virus, an arenavirus related to the virulent human pathogens Junin, Machupo, and Lassa viruses. Infected animals developed decreased activity of multiple coagulation factors, decreased antithrombin III levels, high levels of fibrin-fibrinogen degradation products, impaired platelet function, and thrombocytopenia. Testing for the presence of a coagulation inhibitor revealed a pattern consistent with factor deficiency. Fibrin thrombi were not found at necropsy. The findings of high fibrin-fibrinogen degradation product levels and decreased antithrombin III levels, in association with decreased activity of multiple coagulation factors and thrombocytopenia, suggest that intravascular coagulation is a feature of this experimental infection. The demonstration of abnormal platelet function is also significant, as this could contribute to defective hemostasis despite the moderate thrombocytopenia which usually occurs in arenaviral disease. PMID- 3030152 TI - [Effects of GABA-ergic and glycinergic agents on pentamethylenetetrazole-induced convulsions in rats submitted to repeated injections of reserpine]. PMID- 3030153 TI - Painful ophthalmoplegia caused by metastasis of cholangiocarcinoma of the liver. PMID- 3030151 TI - Prognostic indicators in cystosarcoma phylloides. AB - Cystosarcoma phylloides is a breast neoplasm that has a frequently unpredictable clinical course. We made a retrospective study of 25 patients with this disease in an attempt to evaluate the indicators of aggressive behavior. In our series, older patient age, nulliparity, rapid tumor growth, pain, and large size of tumors increased the suspicion of malignancy but were not always reliable indicators of malignancy. Skin ulceration, tumor necrosis, and infiltrating tumor margins were the most ominous characteristics. High-grade tumors, that is, those with increased cellularity, vascularity, mitotic figure, and pleomorphism, often indicated aggressive behavior. Mixed mesenchymal components were sometimes related to a malignant course. We found a 24 percent incidence of associated breast cancer. Carcinoma of the ipsilateral breast was found in four patients and later in the contralateral breast in two patients. Of our 25 patients, 10 (40 percent) had recurrence and 4 (16 percent) died from disease. Recurrences after treatment usually occurred within 3 years. Patients must be followed carefully for local recurrence or metastases, since the clinical course is not predictable. Forty percent of the lesions were diagnosed as being malignant. Local excision was associated with recurrence in six of eight patients and was clearly inadequate treatment. Quadrantectomy was effective for benign peripheral lesions when a generous margin could be obtained. From these data, we believe that mastectomy is indicated in all patients with malignant lesions and in those with large benign lesions. PMID- 3030154 TI - Administration of ACTH to suckling rats results in hyperkinetic behavior. AB - The present study on ACTH-administered suckling rats was undertaken to monitor the late behavioral sequelae due to the effects of this drug. The locomotor activity of rats treated with ACTH in the suckling period increased significantly as to the gross size of movements when compared with control rats. The increase in locomotor activity was correlated with the suppression of CNPase activity. These results show that administration of ACTH during the suckling period has suppressive effects on the development of the brain and behavior, and that these effects persist for a longer period after termination of the administration of ACTH than the direct effect of the peptide on endocrine functions. PMID- 3030155 TI - Cerebral blood flow and brain shrinkage seen on CT during ACTH therapy. AB - By means of the Doppler ultrasound method, the cerebral blood flow (CBF) was assessed in 21 children with epilepsy undergoing treatment with adrenocorticotrophic hormone (ACTH). The maximum reduction in the internal carotid velocity, as an index of CBF during therapy, was about 35 percent compared with the values before therapy. Furthermore, sequential computed tomography (CT) examinations of the same subjects were performed to evaluate the change in the area of the intracranial brain parenchyma during therapy. The maximum reduction in the parenchymal area during therapy was about 10 percent. This corresponds to a 20 percent reduction in CBF according to Poiseuille's law, however, the remaining reduction in CBF demonstrated by velocity measurement cannot be explained only by that mechanical vascular factor. From these findings, it is concluded that in order to elucidate the mechanism of the CBF reduction, physiological factors such as changes in metabolism during therapy should also be evaluated in addition to the mechanical and physical causes. PMID- 3030156 TI - Silica containing primary hydroxyl groups for high-performance affinity chromatography. AB - Primary hydroxyl groups were incorporated into silica by a four-step reaction procedure which includes modification of the silica surface with gamma glycidoxypropyltrimethoxysilane, leading to an epoxide silica; hydrolysis with acid to yield a diol silica; oxidation of the diol silica with periodate to yield a silica resin with aldehyde functions; and reduction with sodium borohydride to obtain the primary hydroxyl-containing silica. The hydroxyl groups were activated with chloroformates or carbodiimidazole. Proteins were coupled under mild conditions in high yield to these activated silica resins. Columns containing these newly developed silica derivatives were used for the fast and efficient purification of antibodies on antigen-containing silica, as well as for the purification of trypsin on a trypsin inhibitor column (or vice-versa). The effect of pressure on association and dissociation of the affinity complex is discussed. PMID- 3030158 TI - Chemiluminescence probe with Cypridina luciferin analog, 2-methyl-6-phenyl-3,7 dihydroimidazo[1,2-a]pyrazin-3-one, for estimating the ability of human granulocytes to generate O2-. AB - The Cypridina luciferin analog, 2-methyl-6-phenyl-3,7-dihydroimidazo[1,2 a]pyrazin-3-one (CLA), in Hanks' balanced salt solution, emitted a weak luminescence which was not affected by superoxide dismutase or catalase and was not augmented by resting human granulocytes. In contrast, activated granulocytes caused a dramatic increase in the luminescence of CLA. The light emission by CLA in the presence of activated granulocytes was inhibited by superoxide dismutase, but not by catalase or benzoate. Azide at 0.5 mM did not inhibit light emission significantly. These results indicate that O2-, rather than H2O2, HO., singlet oxygen, or HOCl, was the agent responsible for eliciting the chemiluminescence of CLA. Moreover, the intensity of light emission by CLA correlated with the rate of production of O2- either by activated neutrophils or by the xanthine oxidase reaction. PMID- 3030159 TI - A spectrophotometric collagenase assay. AB - A quantitative collagenase assay using Coomassie blue staining and microtiter spectrophotometry is described. Collagen is gelled and dried onto the bottom of microwells as substrate, washed, incubated with samples, washed again, and then stained. Absorbance at 590 nm increases linearly with increasing amounts of collagen in the range 5-40 micrograms. Bacterial and mammalian collagenases can be detected within 2 h, and 10 ng of bacterial collagenase may be detected in 16 h. For simple screening applications, activity may be detected by eye. The assay is safe, simple, fast, economical, and sensitive. PMID- 3030157 TI - High resolution preparative gel electrophoresis of DNA fragments and plasmid DNA using a continuous elution apparatus. AB - An apparatus designed for preparative gel electrophoretic separation of proteins (M. A. Hediger, (1984) Anal. Biochem. 142, 445-454) has been used successfully for separating DNA restriction fragments. The apparatus displayed yields and resolutions that are higher than those obtainable with commercially available devices. The amounts of DNA applied to the column range from a few micrograms to milligram quantities. Restriction DNA fragments very similar in size were isolated in pure form with the apparatus. After ethanol precipitation, these fragments were successfully used for restriction enzyme cleavages, ligation, or chemical sequencing. Furthermore the apparatus provides a convenient method for the large-scale isolation of plasmid DNA. The method requires only 4 h of electrophoresis and therefore greatly reduces the preparation time compared with the conventional equilibrium centrifugation method which requires centrifugation times of up to 60 h. In contrast to the centrifugation method, contaminants such as RNA, proteins, and chromosomal DNA are efficiently removed by this technique. PMID- 3030160 TI - Accelerated reversal of atracurium blockade with priming doses of edrophonium. PMID- 3030161 TI - Premedication abolishes the increase in plasma beta-endorphin observed in the immediate preoperative period. PMID- 3030162 TI - Platelet resistance to prostacyclin. Enhancement of the antiaggregatory effect of prostacyclin by pentoxifylline. AB - With regard to existence of high prostacyclin (PGI2) levels during atheromatosis and thrombus formation, resistance of platelets to prostacyclin and its analogues seems to play an important pathophysiologic key role for the clarifying of vasoocclusive phenomena. Platelet resistance to prostacyclin was studied in vitro and ex vivo in 160 atherosclerotic patients (assessed by objective diagnostic criteria) with and without thrombotic complications and in 50 controls. Prostacyclin resistance phenomena were more pronounced and frequent in patients with occlusive complications, the difference from controls being statistically significant. However, there was no significant difference between the controls and the nonthrombotic patient sample. The intraplatelet cAMP levels might be the metabolic basis of the PGI2 resistance phenomenon, because in the patient group, platelet cAMP levels were decreased by 50% after Ca2+ stimulation. Compared to controls beta-thromboglobulin and thromboxane B2 plasma levels were significantly increased (30 +/- 9 to 87 +/- 26 ng/ml and 9 +/- 5 to 54 +/- 21 pg/ml, respectively), confirming the hyperreactivity state of resistant platelets. From the therapeutic point of view, patients with resistant platelets require PGI2 doses that cause, however, increased side effects. We were able to demonstrate in vivo that IV pretreatment with pentoxifylline--a known stimulator of cAMP formation in platelets--followed by a simultaneous and continuous IV infusion of PGI2 + pentoxifylline, permitted us to reduce significantly the mean PGI2 doses needed for triggering an antiplatelet effect, without inducing side effects. In ex vivo studies, PGI2 resistant platelets of atherosclerotic patients pretreated with pentoxifylline showed normalized stimulation response, and platelet cAMP levels increased from 7.8 +/- 2.7 to 15.2 +/- 1.9 pmol/10(8) platelets.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3030163 TI - [Anesthesia for surgery of insulinoma]. AB - The difficulties of interpretation of blood sugar level changes during the postoperative period in the anaesthetic management of insulinoma are discussed. Several specific means reduced the errors in the assessment of the hyperglycemic rebound which occurred after the removal of the tumour. They consisted of continuous sugar infusion accorded to measured glucose levels, in order to maintain a constant blood sugar value between 50 and 70 mg X 100 ml-1 before removal of the insulinoma. Furthermore, analgesia was provided by high doses fentanyl. Sugar containing solutes were avoided during the procedure. Glucose levels rose slowly after tumour removal and reached 170 mg X 100 ml-1 at 120 min. This rebound was known to be of no help in ascertaining complete resection. Simultaneous determinations of blood glucose and insulin were obtained. The value of portal blood insulin was found to be normal (12.3 mU X l-1) 30 min after insulinoma removal. Turner's index calculated every 30 min decreased simultaneously (143) and reached a normal value at 120 min (39). These results, obtained during the surgical procedure all the more easily because of rapid laboratory procedures, could be better arguments in determining whether tumour removal has been complete. PMID- 3030164 TI - Energy metabolism in canine erythrocytes associated with inherited high Na+- and K+-stimulated adenosine triphosphatase activity. AB - Energy metabolism in canine erythrocytes associated with inherited high Na+- and K+-stimulated adenosine triphosphatase [(Na,K)-ATPase] activity (HK cells) was compared with that in normal canine erythrocytes (LK cells). Activities of some of the glycolytic enzymes in the HK cells were significantly higher than those in LK cells. The concentrations of adenosine triphosphate (ATP) and glycolytic intermediates in HK cells were almost equal to those in LK cells. Glucose utilization and lactate production by HK cells in vitro and incorporation of [32P]orthophosphate or [14C]glucose into 2,3-diphosphoglycerate in HK cells were higher than in LK cells. Radioactivity of [32P]ATP in HK cells was lower than in LK cells, but increased to approximately that of LK cells when (Na,K)-ATPase of HK cells was completely blocked by ouabain. When HK cells and LK cells were incubated in the absence of glucose, the concentration of ATP in HK cells was decreased more than that of LK cells. Although ouabain reduced the rate of decrease in ATP in HK cells, the decrease in ATP in HK cells was still 2-fold that in LK cells. The half-life of HK cells was about one-half that of LK cells. The results indicated that glycolysis is greater in HK cells than in LK cells, and that the increased glycolysis in HK cells was stimulated by an increased rate of ATP breakdown for active cation transport by the (Na,K)-ATPase and by increased degradation of ATP for some other pathway, eg, glutathione synthesis. Thus, the increased demand for ATP in HK cells might result in shortening the lifespan of HK erythrocytes. PMID- 3030165 TI - Effects of feline leukemia virus infection on neutrophil chemotaxis in vitro. AB - A procedure for measuring in vitro feline neutrophil chemotaxis was developed, using a modified Boyden chamber apparatus and 3-microns-pore polycarbonate filters. A pooled feline serum sample was used as the chemoattractant. Chemotaxis was evaluated in 5 groups of cats: group 1-specific-pathogen-free cats that had not been exposed to feline leukemia virus (FeLV); group 2-previremic, FeLV infected, specific-pathogen-free cats; group 3-FeLV-viremic, subclinically affected cats; group 4-FeLV-viremic, clinically affected cats; and group 5-sick cats that were not infected with FeLV. Neutrophils from the viremic, clinically affected cats had significantly lower (P less than 0.025) chemotactic responses than did those from subclinically affected, viremic cats. Conversely, neutrophils from cats that were ill due to causes other than FeLV had the highest mean chemotactic values. Among the viremic, subclinically affected cats, a linear relationship was found between age and chemotaxis, indicating that impairment of neutrophil function may be greater in younger viremic cats. However, FeLV infected cats can not be identified on the basis of neutrophil chemotaxis. PMID- 3030166 TI - Relationships of bovine leukemia virus prevalence in dairy herds and density of dairy cattle to human lymphocytic leukemia. AB - A case-control study was conducted to examine possible relationships between human acute lymphoid leukemia and exposure to dairy cattle and drinking of raw milk. Two hundred twenty-three persons with acute lymphoid leukemia, diagnosed during the years 1969 to 1971 and 1973 to 1980 from the 87 most rural Iowa counties, were accessed from case records at the Iowa State Health Registry for participation in the present study. Each person and 2 matched controls were interviewed for history of residence, exposure to dairy cattle, and consumption of nonpasteurized dairy products. Two types of comparisons between affected persons and controls were done: the prevalence of bovine leukemia virus infection (as measured by serologic study) in dairy herds with which the affected persons and controls had either occupational contact or from which they had consumed raw milk and the density of dairy cattle in the townships where affected persons and controls lived. The bovine leukemia virus infection prevalence in dairy herds with which affected persons had contact was 20%, whereas the infection prevalence in the herds with which the controls had contact was 38%. The density of dairy cows in townships where affected persons resided was generally less than that in townships where controls resided. However, there was one exception; the density of dairy cows at 20 years before diagnosis was higher (589) in townships where affected adult female persons resided, compared with that in townships where controls resided (567). PMID- 3030167 TI - Distribution and implications of beta-endorphin and ACTH-immunoreactive cells in the intermediate lobe of the hypophysis in healthy equids. AB - The distribution of cells that stain positive for beta-endorphin and ACTH immunoreactivity was studied in the pars intermedia (PI) of the hypophysis in 3 healthy horses and 2 healthy ponies. Serial sections treated with commercial antibodies generated against beta-endorphin or ACTH were processed for immunocytochemical studies, using the avidin biotin immunoperoxidase-complex method. Distribution patterns of cells reacting with antibodies were similar in cells from all equids. Cells immunostained for ACTH were numerous and widely distributed in the PI. Cells immunopositive for ACTH probably contain corticotrophin-like intermediate lobe peptide that cross-reacts with antisera to ACTH. Cells immunopositive for beta-endorphin were fewer in number and had a more limited distribution in the PI. Most beta-endorphin-positive cells were located along the border of the PI adjacent to the lobus nervosus and had abundant eosinophilic cytoplasm when stained with hematoxylin and eosin. Cells with prominent eosinophilic cytoplasm were not common in other areas of the PI. When serial sections were examined, cells that stained positive for beta-endorphin immunoreactivity also appeared positive for ACTH. PMID- 3030168 TI - The prone position improves arterial oxygenation and reduces shunt in oleic-acid induced acute lung injury. AB - The arterial oxygen tension (PaO2) may increase when patients with the adult respiratory distress syndrome are turned from supine to prone. We sought to reproduce this observation in dogs with acute lung injury to study the physiologic mechanism by which the improvement in oxygenation might occur. Twenty anesthetized dogs were ventilated with a constant tidal volume (20 ml/kg) of 100% oxygen. Oleic acid (0.09 ml/kg) was injected into the right atrium while rotating the animals through 360 degrees in 4 stages. Animals in Group I (n = 5) remained supine for 10 to 120 min until the supine PaO2 fell below 200 mm Hg. Those in Group II (n = 4) were kept prone during this period. Dogs in Groups I and II were then turned supine or prone every 30 min 5 times. Cardiac output and pulmonary vascular pressures, functional residual capacity (helium dilution), and regional diaphragmatic motion (determined by dorsal and ventral diaphragmatic markers relative to markers on the chest wall seen on lateral chest radiographs taken at FRC and at end-inspiration) were obtained in each position. Eleven dogs were kept supine (Group III, n = 6) or prone (Group IV, n = 5) for 2 h after oleic acid infusion, after which intrapulmonary shunt (Qs/QT) and ventilation-perfusion heterogeneity were measured in the supine and prone positions using the multiple inert gas elimination technique.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3030169 TI - Eosinophil activation in the lung is related to lung damage in adult respiratory distress syndrome. AB - The concentrations of eosinophil cationic protein (ECP), a specific granule constituent of eosinophil granulocytes, were measured in bronchoalveolar lavage (BAL) fluid from patients with adult respiratory distress syndrome (ARDS). The mean level was 163 +/- 85 (SD) micrograms/L and significantly increased (p less than 0.001) compared with the levels in control subjects (19 +/- 18 micrograms/L). The BAL fluid concentrations of myeloperoxidase (MPO) were also significantly increased in ARDS, indicating a local neutrophil activation. A significant correlation was found between BAL fluid ECP and MPO, suggesting a common activator of eosinophils and neutrophils. No relation was seen between BAL fluid ECP or MPO and degree of complement consumption, suggesting that other mechanisms besides complement activation may underlie granulocyte activation in ARDS. Lavage ECP levels correlated strongly with the severity of lung damage defined by pulmonary oxygenation index (PaO2/inspired fraction of O2). Activation of eosinophils, neutrophils, and the complement system is not specific for ARDS but was also observed in patients after major surgery, but at a significantly lower level (p less than 0.001). It is concluded that eosinophil activation is part of the inflammatory process in the lung in ARDS. The association observed between elevated lavage fluid levels of ECP and reduced pulmonary function in ARDS might reflect a pathophysiologic role for ECP and other cytotoxic eosinophil products in this syndrome and should be evaluated. PMID- 3030170 TI - Postirradiation malignant fibrous histiocytoma of the trachea. AB - A 77-yr-old woman presented with a malignant fibrous histiocytoma (MFH) of the trachea 11 yr after right subtotal thyroidectomy and delivery of 5,400 rads to the neck for infiltrating papillary thyroid carcinoma. The tumor developed in the irradiated area. Postirradiation MFH tends to occur in subcutaneous tissues, and is rare in lung parenchyma or airways. PMID- 3030171 TI - Wilms' tumors in adults. AB - Four cases of adult Wilms' tumor are reported comparing their diagnoses, treatment, and survival with the world literature. The oldest recorded patient is herein described at age 84. Aggressive surgical and medical therapy is necessary to improve survival. Surgical exploration should be through a transperitoneal approach with the patient positioned such that access to the chest can be obtained for maximum exposure and resection. Chemotherapy consisting of actinomycin D and vincristine should be instituted in all patients. Radiation therapy must also be considered in any patient with the diagnosis of nephroblastoma. PMID- 3030172 TI - Granular cell tumors. AB - A lesion of unknown etiology and histogenesis, the granular cell tumor usually arises in the skin or soft tissue. It has been reported, however, in other sites and can be multifocal. The authors have seen 31 such tumors in 26 patients in their institutions since 1970. Most (21) patients were females, and 12 patients were black. The average age was 41.8 years, excluding two newborns with a congenital granular cell myoblastoma of the gingiva. The most common site of occurrence was the skin; seven tumors originated from the trunk and five from the extremities. Four lesions were found in the breast, three on the vulva, two in the axilla, two in the gum, two in the buccal cavity, two in the esophagus, and one each in the stomach, gluteus muscle, eyelid, and bronchus. Two patients had multiple synchronous lesions. These were bilateral hand lesions in one patient, and lesions of the breast and axilla in the other. A third patient had three separate lesions which arose over the course of 5 years, involving the bronchus, the gluteus muscle, and the buccal mucosa. An additional esophageal lesion was an incidental finding 3 years after excision of a granular cell tumor of the breast. All of the tumors were removed with local, simple excision, except for the 2-cm lesion in the stomach for which a wedge resection of the fundus was necessary and the bronchial lesion for which a wedge resection of the left upper lobe of the lung was performed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3030173 TI - Fish-oil fatty acid supplementation in active rheumatoid arthritis. A double blinded, controlled, crossover study. AB - STUDY OBJECTIVE: to determine the efficacy of fish-oil dietary supplements in active rheumatoid arthritis and their effect on neutrophil leukotriene levels. DESIGN: nonrandomized, double-blinded, placebo-controlled, crossover trial with 14-week treatment periods and 4-week washout periods. SETTING: academic medical center, referral-based rheumatology clinic. PATIENTS: forty volunteers with active, definite, or classical rheumatoid arthritis. Five patients dropped out, and two were removed for noncompliance. INTERVENTIONS: treatment with nonsteroidal anti-inflammatory drugs, slow-acting antirheumatic drugs, and prednisone was continued. Twenty-one patients began with a daily dosage of 2.7 g of eicosapaentanic acid and 1.8 g of docosahexenoic acid given in 15 MAX-EPA capsules (R.P. Scherer, Clearwater, Florida), and 19 began with identical appearing placebos. The background diet was unchanged. MEASUREMENTS AND MAIN RESULTS: the following results favored fish oil placebo after 14 weeks: mean time to onset of fatigue improved by 156 minutes (95% confidence interval, 1.2 to 311.0 minutes), and number of tender joints decreased by 3.5 (95% Cl, -6.0 to 1.0). Other clinical measures favored fish oil as well but did reach statistical significance. Neutrophil leukotriene B4 production was correlated with the decrease in number of tender joints (Spearman rank correlation r=0.53; p less than 0.05). There were no statistically significant differences in hemoglobin level, sedimentation rate, or presence of rheumatoid factor or in patient reported adverse effects. An effect from the fish oil persisted beyond the 4-week washout period. CONCLUSIONS: fish-oil ingestion results in subjective alleviation of active rheumatoid arthritis and reduction in neutrophil leukotriene B4 production. Further studies are needed to elucidate mechanisms of action and optimal dose and duration of fish-oil supplementation. PMID- 3030174 TI - Epstein-Barr virus infections in males with the X-linked lymphoproliferative syndrome. AB - A registry of persons with the X-linked lymphoproliferative syndrome, which is characterized by marked susceptibility to diseases induced by the Epstein-Barr virus, has enrolled 161 patients within 44 kindreds. Fifty-seven percent of the males died of infectious mononucleosis, 29% developed acquired hypogammaglobulinemia, and 24% had malignant lymphoma. The mortality rate was 80%; 70% died by 10 years of age and 100% by 40 years. Thirty-two boys survive, most with malignant lymphoma, acquired hypogammaglobulinemia, or both. We hypothesized that the defective lymphoproliferative control locus on the X chromosome results in unregulated cytotoxic lymphocytic responses to the Epstein Barr virus; hence, severe hepatitis and virus-associated hemophagocytic syndrome occur with the infectious mononucleosis phenotype. T-cell suppression of immunoglobulin secretion by B cells is responsible for acquired hypogammaglobulinemia. A sustained polyclonal B-cell proliferation probably converts to a monoclonal B-cell malignancy as a result of molecular alterations. PMID- 3030175 TI - Cytomegalovirus thrombophlebitis after successful DHPG therapy. PMID- 3030176 TI - [Intestinal bacterial toxins and hepatitis]. PMID- 3030177 TI - [Intestinal bacterial flora and tumors of the colon: intestinal excretion and degradation of neutral steroids in subjects at risk for cancer of the colon]. PMID- 3030178 TI - [Cellular bases of hypertension in pregnancy. I. Electrolytes and their regulation. Review of the literature and methodological introduction to the study]. PMID- 3030179 TI - [Magnetic resonance imaging of craniofacial pathology]. AB - Effectiveness of nuclear magnetic resonance imaging (MRI) was compared with that of the CT scan in several cases involving different lesions: intracranial, base of skull, facial sinuses, parapharyngeal spaces. MRI provided excellent contrast in soft tissues with good detection of tumors and their limits, the condition of neighboring tissues and any local or regional tumor extension. Vessels were also visualized. It was not influenced by dental artefacts, and allowed three dimensional sections to be obtained simply without manipulation of patients. In contrast, bone structures were analyzed less clearly than by the CT scan. Marked progress in MRI technical features and results obtained can be expected in the very near future. PMID- 3030180 TI - [Male genital lesions caused by papillomaviruses. Importance of colposcopy]. AB - Ten years after the description of cervical flat condyloma, it is now admitted that Human Papillomaviruses (HPVs) type 6 and 11 are responsible for condylomata acuminata and typical flat condyloma of the uterine cervix. HPV DNA type 16 and, less frequently, 18, 33 and other as yet uncharacterized HPV types (G. Orth, personal communication), are found in the majority of Cervical Intraepithelial Neoplasia (CIN), Vulvar Intraepithelial Neoplasia (VIN) and cervical and vulvar invasive cancers. Since HPVs are sexually transmissible, recent interest has focused on the "male factor". Clinically detectable lesions of male genitalia are condylomata acuminata, bowenoid papulosis and flat condyloma-like papules. The aim of our study was the colposcopical screening, recently suggested, of different groups of male patients in order to detect HPV-related lesions and the description of the colposcopical features of subclinical HPV-related lesions, since most of them have never been reported, to our knowledge, in the literature. A total of 114 men were examined. Among them, 18 presented clinically detected recalcitrant condylomata acuminata, 28 had been treated for the same pathology 1 to 5 days earlier and were clinically free of lesions, 46 were sexual partners of women with cervical atypia (flat condyloma and/or CIN) and 22 had a clinical diagnosis of genital infection without HPV-related lesions. A careful examination of external genitalia was performed. Then all patients underwent penile colposcopy before and after application of 5 p. 100 acetic acid. Selected biopsies were performed in all lesions which were clinically and colposcopically different from classic warts. Colpophotographs were taken in all cases.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3030181 TI - Adrenergic nerves and the delay phenomenon. AB - The present study indicates that adrenergic nerves can in part explain the delay phenomenon. Data presented suggest that degeneration in release of norepinephrine might be deleterious to critical parts of a skin flap. This could be avoided by performing a delay procedure, as no release of norepinephrine will occur after the second operation. PMID- 3030183 TI - [Action of various disinfectants on 7 envelope-free viruses]. AB - The activity of 15 disinfectants on seven nude viruses belonging to groups significant in human and veterinary medicine and in the bio-industry was investigated by a dilution technique. These trials, performed organic matters being either present or not revealed that current products such as 0.8% sodium hydroxyde and 1 degree chlorine water were the most active. According to their compositions, the chemicals varied greatly in their efficacy. PMID- 3030184 TI - Carcinoembryonic antigen (CEA) and gastrointestinal cancer associated antigen CA 19-9 in bronchioloalveolar carcinomas and pulmonary adenocarcinomas. AB - The concentrations of carcinoembryonic antigen (CEA) and the gastrointestinal cancer associated antigen CA 19-9 were studied by radioimmunoassay in serum samples from patients with malignant and benign pulmonary tumours. Elevated levels of either markers were found in 14% of patients with malignant tumours and in none of the cases with benign lesions. Following removal of the tumour, a decline in high CA 19-9 levels was found. The advanced tumours produced the highest serum CA 19-9 concentrations but elevated values were also found in small bronchioloalveolar carcinomas and pulmonary adenocarcinomas. Immunohistochemical staining for CEA and CA 19-9 was performed on routine histopathological specimens. About half of the cases of bronchioloalveolar carcinomas and pulmonary adenocarcinomas were CA 19-9 positive and about 90% were CEA positive. No correlation between intensity of tissue expression and serum levels of CA 19-9 could be demonstrated. However, the bronchioloalveolar carcinomas and pulmonary adenocarcinomas which produced high serum levels of CA 19-9 also stained positively. PMID- 3030182 TI - Study of the efficacy of an inactivated virus vaccine against porcine parvovirus. AB - The efficacy of an inactivated virus vaccine against porcine parvovirus has been studied by immunizing 4 sows during pregnancy. A parvovirus virulent strain has been inoculated to these sows and to two other unvaccinated sows used as controls. The infection was performed between the 52nd and the 57th day of gestation. In the litters born from the vaccinated sows, 82% of the piglets were alive and normal. Neither PPV antibodies nor antigen could be revealed in the stillborn fetuses born from the 4 vaccinated sows. Reversely, only 9.5% of the piglets born from the 2 unvaccinated sows were alive at birth, although they were probably infected during pregnancy. In total, 86% of fetuses in these 2 litters were mummified. A field study allowed to show that the double vaccination antibodies induced, persisted with constant titers for, at least, 13 months. Moreover, the reproductive performance of 413 gilts, vaccinated twice before mating, were not affected by this treatment. PMID- 3030185 TI - [Influence of heparin on the radioimmunological assay of ACTH]. AB - Heparin traps plasma ACTH, promoting the formation of aggregates with apparent high molecular weight as shown by chromatography on Sephadex G 50 fine columns. The percentage of 125I-ACTH which appeared in the void volume of the column, increased linearly with the log. dose of heparin. Heparin at concentrations of up to 100 IU/ml does not impair ACTH adsorption on either silicic acid or Quso G 32 as well as further elution by acetic acid/acetone/water (I : 40 : 59; V/V) or HCl O.I N. Silicic acid traps selectively ACTH but not heparin. Heparin interferes with direct RIA-ACTH in the plasma by decreasing 125I-ACTH binding to the antibodies and modifying the slope of the standard curve. Unsuitable artefacts induced by heparin, as overestimation or underestimation of plasma ACTH levels by RIA, can be avoid by previous hormone extraction from heparinized plasmas. Such results emphasized the importance of the sample preparation in order to obtain consistent results. PMID- 3030186 TI - Chronic inflammatory demyelinating polyradiculoneuropathy associated with pregnancy. AB - In a series of 61 patients with the relapsing variety of chronic inflammatory demyelinating polyneuropathy, there were 16 women of childbearing age, 9 of whom became pregnant. In 4 of these women, the onset of neuropathy occurred in pregnancy and in the other 5 relapses occurred during pregnancy. There was a significant increase in the number of relapses during the year of pregnancy, and a tendency for symptoms to worsen during the third trimester or immediate postpartum period. It is concluded that there is an increased risk of relapse of chronic inflammatory demyelinating polyneuropathy in pregnancy. PMID- 3030187 TI - Primary amyloid polyneuropathy with myocardial uptake of technetium-99m diphosphonate. PMID- 3030188 TI - Inflammatory demyelinating peripheral neuropathies associated with human T-cell lymphotropic virus type III infection. AB - Nine patients with inflammatory demyelinating polyneuropathies (IDP) were found to have human T-cell lymphotropic virus type III (HTLV-III) infection. The 8 men, 6 of whom were homosexual, and 1 woman, a former intravenous drug user, presented with progressive weakness. Two had lymphadenopathy but all were otherwise asymptomatic. Six had chronic IDP and 3 had Guillain-Barre syndrome. In addition to an elevated cerebrospinal fluid (CSF) protein level (mean, 193 mg/dl), most patients had cerebrospinal fluid pleocytosis (mean, 23 cells/mm3), a distinctive feature. All had reduced T4:T8 T-cell ratios. Results of nerve conduction studies were characteristic of demyelination. Nerve biopsies revealed intense inflammatory cell infiltrates and macrophage-mediated demyelination. The patients recovered either spontaneously or following treatment with corticosteroids or plasmapheresis. During a mean interval of 20 months after presentation, only 1 patient had developed acquired immune deficiency syndrome. Patients with HTLV-III infection have disordered immune function, and the mechanism of the development of the IDP is likely to be immunopathogenic. As a result of our experience, we suggest that all patients with IDP be tested for evidence of HTLV-III infection. We also found, although in uncontrolled trials, that treatment with either prednisone or plasmapheresis was followed by clinical improvement; since plasmapheresis is not likely to further depress cell-mediated immunity, we suggest that it be the initial therapy. PMID- 3030189 TI - Type 1 human poliovirus binds to human synaptosomes. AB - Poliovirus is a neurotropic virus that selectively infects human motoneurons in vivo. The basis for the specificity of this infection is not fully understood. It has been suggested that this tropism occurs because motoneurons are the only neurons to express poliovirus receptors. We have examined this hypothesis by measuring the binding of 125I-labeled poliovirus type 1 to neural tissues. With this assay we have detected highly specific binding sites in human but not rodent neural tissue. Regional assays of binding in human central nervous system homogenates demonstrate that binding sites are not confined to motoneurons. Rather, they are widely distributed throughout the human neuraxis. Particularly in the forebrain, binding is more abundant in gray than white matter. For this reason, we performed tissue fractionation studies which indicate that poliovirus binding sites are enriched in synaptosomes, the subfraction of central nervous system gray matter tissue rich in synaptic endings. The preferential expression of poliovirus binding sites in synaptic endings may be an important factor in the motor tropism of this virus, inasmuch as the major category of neurons with synaptic endings outside the central nervous system are motoneurons; thus, particles of virus may preferentially bind to this cell type during poliovirus viremia. PMID- 3030190 TI - Endemic tropical spastic paraparesis associated with human T-lymphotropic virus type I: a clinical and seroepidemiological study of 25 cases. AB - Tropical spastic paraparesis (TSP) is a common myeloneuropathy with primary and predominant involvement of the pyramidal tract and minimal sensory loss. The epidemic form of TSP is related to toxic nutritional factors, but the endemic form occurs in clusters in tropical areas, especially in India, Africa, the Seychelles, Colombia, and areas of the Caribbean. We describe the clinical and epidemiological features of 25 TSP patients from Martinique (French West Indies) with serum antibodies to human T-lymphotropic virus type I (HTLV-I). Furthermore, all 11 patients who were seropositive for HTLV-I had specific HTLV-I antibodies in their CSF. All were women. The age of onset varied from 25 to 60 years (mean, 45 years). The main clinical features are spastic paraparesis or paraplegia with spasticity of the upper limbs, minimal sensory loss, and bladder dysfunction. Minimal estimated incidence and prevalence are 1 per 100,000 inhabitants per year and 8 per 100,000, respectively. Seventeen percent of the relatives of patients with HTLV-I-associated TSP have HTLV-I antibodies (1 husband and 7 children). In Martinique, the prevalence of HTLV-I antibodies in the general population is about 2% and reaches 10% for neurological disorders other than TSP. Since our initial report, the association between spastic paraparesis and HTLV-I has been confirmed in Jamaica, Colombia, and Japan, suggesting the neurotropism of this lymphotropic human retrovirus. PMID- 3030191 TI - Association of human T-lymphotropic viruses in chronic neurological disease. PMID- 3030192 TI - Plasmid transformation of Streptomyces tendae after heat attenuation of restriction. AB - Streptomyces tendae ATCC 31160 produces nikkomycin, a fungicide and insecticide that inhibits chitin synthases. Exposure of S. tendae protoplasts to 50 degrees C for 30 min is required for transformation (10(2) thiostrepton-resistant transformants micrograms of DNA-1) with plasmid pIJ702 or pIJ680 from Streptomyces lividans. pIJ702 and pIJ680 DNA isolated from the S. tendae transformants is efficient (10(6) to 10(7) transformants micrograms of DNA-1) in subsequent transformations of S. tendae protoplasts generated at 30 degrees C. PstI fails to cut the single PstI site in pIJ702 and cuts only one of the two PstI sites in pIJ680 DNA isolated from S. tendae transformants. Digests of plasmid DNA mixtures showed that plasmid DNA from S. tendae does not inhibit PstI activity. pIJ702 and pIJ680 DNA from S. tendae transformants was used to transform S. lividans to show that plasmid DNA remains unchanged, except for modification at some PstI sites in S. tendae, as a consequence of passage through S. tendae. The DNA modification is lost when S. lividans is transformed with plasmid DNA from S. tendae transformants. Since S. tendae modifies only some PstI sites, it appears the modification (presumably restriction activity also) activity in S. tendae recognizes a sequence that includes or overlaps the PstI hexanucleotide recognition sequence. PMID- 3030193 TI - Seasonal changes in the cecal microflora of the high-arctic Svalbard reindeer (Rangifer tarandus platyrhynchus). AB - The dominant cecal bacteria in the high-arctic Svalbard reindeer were characterized, their population densities were estimated, and cecal pH was determined in summer, when food quality and availability is good, and in winter, when it is very poor. In summer the total culturable viable bacterial population was (8.9 +/- 5.3) X 10(8) cells ml-1, whereas in winter it was (1.5 +/- 0.7) X 10(8) cells ml-1, representing a decrease to 17% of the summer population density. Of the dominant species of cultured bacteria, Butyrivibrio fibrisolvens represented 23% in summer and 18% in winter. Streptococcus bovis represented 17% in summer and 5% in winter. Bacteroides ruminicola represented 10% in summer and 26% in winter. In summer and winter, respectively, the proportion of the viable population showing the following activities was as follows: fiber digestion, 36 and 48%; cellulolysis, 10 and 6%; xylanolysis, 33 and 48%; and starch utilization, 77 and 71%. The most abundant cellulolytic species in summer was Butyrivibrio fibrisolvens, representing 62% of the total cellulolytic population, and in winter it was Ruminococcus albus, representing 80% of the total cellulolytic population. The most abundant xylanolytic species in summer was Butyrivibrio fibrisolvens, and in winter it was Bacteroides ruminicola, representing 59 and 54% of the xylanolytic isolates in summer and winter, respectively. The cecal bacterial of the Svalbard reindeer have the ability to digest starch and the major structural carbohydrates of the diet that are not digested in the rumen. The cecum in these animals has the potential to contribute very substantially to the digestion of the available plant material in both summer and winter. PMID- 3030195 TI - [HBV and hepatocellular carcinoma]. AB - Significant research evidence has demonstrated an association between persistent infection with hepatitis B virus (HBV) and the generation of hepatocellular carcinoma (HCC). These findings are based on epidemiologic studies, molecular studies and studies of HBV like viruses. Epidemiologically, the geographic correlation between HBV infection and HCC, serum HBsAg in patients with HCC, familial clustering of HCC, prospective studies, and pathological studies are discussed. Molecular studies of HBV, the structure of HBV DNA as related to retroviruses and integration of HBV DNA into the host DNA are demonstrated. The structure and replication of HBV are somewhat similar to those of retroviruses. The incidence of HBV DNA integration into the host chromosome of the patients with HBV infection is high. The structure of integrated HBV DNA sequences and flanking sequences was analyzed in many cases. However, none of the classical mechanisms of viral oncogenesis has thus far been demonstrated. The role of HBV in HCC is not understood at the molecular level. HBV may act as just an initiator, or HBV DNA integration may have an active role in liver cancer. The woodchuck hepatitis virus (WHV) is the most oncogenic and suitable animal model. Using this model, we show some results of our experiments. PMID- 3030194 TI - Influence of dietary fiber on xylanolytic and cellulolytic bacteria of adult pigs. AB - Xylanolytic and cellulolytic bacteria were enumerated over an 86-day period from fecal samples of 10 8-month-old gilts that were fed either a control or a 40% alfalfa meal (high-fiber) diet. Fecal samples were collected from all pigs on days 0, 3, 5, 12, 25, 37, 58, and 86. Overall, the numbers of xylanolytic bacteria producing greater than 5-mm-diameter zones of clearing on 0.24% xylan roll tube medium after 24 to 36 h of incubation were 1.6 X 10(8) and 4.2 X 10(8)/g (dry weight) of feces for the control pigs and those fed the high-fiber diet, respectively. After 1 week of incubation, a large number of smaller zones of clearing (1 to 2 mm) appeared. Besides Bacteroides succinogenes and Ruminococcus flavefaciens, which produced faint zones of clearing in xylan roll tubes, three strains which closely resembled B. ruminicola hydrolyzed and used xylan for growth. The overall numbers of cellulolytic bacteria producing zones of clearing in 0.5% agar roll tube medium were 0.36 X 10(8) and 4.1 X 10(8)/g for the control pigs and those fed the high-fiber diet, respectively. B. succinogenes was the predominant cellulolytic isolate from both groups of pigs, and R. flavefaciens was found in a ratio of approximately 1 to 15 with B. succinogenes. Degradation of xylan and cellulose, measured by in vitro dry matter disappearance after inoculation with fecal samples, was significantly greater for pigs fed the high-fiber diet than that for the controls. These data suggest that the number of fibrolytic microorganisms and their activity in the large intestine of the adult pig can be increased by feeding pigs high-alfalfa-fiber diets and that these organisms are similar to those found in the rumen. PMID- 3030196 TI - [Analysis of the signalling pathway of TNF in normal cells and tumor cells]. AB - The internalization process and intracellular distribution of 125I-labeled TNF, in L-M (murine tumorigenic fibroblasts, highly sensitive to TNF cytotoxicity) cells and in HEL (human embryonic lung cells, non-sensitive to TNF cytotoxicity) cells bearing TNF receptor, were elucidated by pulse-chasing and by Percoll density gradient centrifugation. Effect of TNF treatment on the RNA and protein synthesis of target cells was also studied using 3H-UDR and 35S-methionine incorporation. In both L-M and HEL cells, receptor-bound 125I-TNF was rapidly internalized and delivered to lysosomes within 15-30 min, followed by degradation and release into the culture medium. RNA synthesis and protein synthesis were not affected by TNF treatment in HEL cells, but marked stimulation (3.5 times and 4.2 times, respectively) was observed in L-M cells. PMID- 3030197 TI - [Treatment of various malignancies with recombinant IFN-gamma (S-6810). The IFN gamma Study Group]. AB - Recombinant interferon-gamma (rIFN-gamma) was given to patients with malignancies by continuous daily administration or by intermittent high doses. Local administration was performed for patients with skin malignancy, and bladder and hepatic carcinoma. Among 239 cases eligible for evaluation (144 cases treated by systemic administration and 95 cases by local injection), complete response was observed in one case each of renal cell carcinoma, malignant lymphoma and mycosis fungoides, all treated by systemic administration. Intermittent high doses of rIFN-gamma induced a response rate of 21.4% in renal cell carcinoma. This rate was higher than the 8.6% obtained following continuous administration. Average response rate to local injection of rIFN-gamma in skin malignancies was 55.3%. This value was comparable with that obtained by IFN-alpha treatment. Fever was observed in 89% of the cases treated by systemic administration. Chills, malaise and anorexia were the main side effects. Local injection also induced similar side effects, although the incidence was lower in comparison with systemic administration. Incidence of side effects was higher in the intermittent high dose group than in the continuous administration group for all items except fever. However, patients showed good tolerance to high doses of rIFN-gamma reaching 40 X 10(6) U/m2/day. PMID- 3030198 TI - [Clinical evaluation of serum carbohydrate antigen 19-9 in carcinoma of the lung- a comparison with carcinoembryonic antigen]. AB - Serum levels of carbohydrate antigen 19-9 (CA 19-9) were measured in 235 untreated patients with lung cancer, 20 patients with benign pulmonary disease and 39 healthy controls. In almost all these patients, carcinoembryonic antigen (CEA) was determined at the same time. The positivity of CA 19-9 in lung cancer patients was significantly higher than in those with benign pulmonary disease (30.2% vs. 5%: P less than 0.05). With regard to histological types of lung cancer cases, the positivity of CA 19-9 in adenocarcinoma cases was higher than that in cases of squamous cell carcinoma (39.6% vs. 16.4% : P less than 0.01). With regard to clinical stages of lung cancer positivity of CA 19-9 in stage IV (37.9% vs. 10.3% : P less than 0.01) or stage III (30.3% vs. 10.3% : P less than 0.05) was significantly higher than in stage I. Sensitivity, specificity and accuracy in the detection of lung cancer were 31%, 95%, and 36% for CA 19-9, and 46%, 80% and 49% for CEA, respectively. Combined evaluation with CA 19-9 and CEA gave the highest specificity (98%) when both were measured, and the highest sensitivity (55%) when at least one was measured. Combined assessment of CA 19-9 and CEA improved the measurement of each one in the detection of lung cancer. PMID- 3030199 TI - [Cancer of the testis: our experience in 177 cases]. PMID- 3030200 TI - Adenoid cystic carcinoma of the breast. Data from the Connecticut Tumor Registry and a review of the literature. AB - The case records of the Connecticut Tumor Registry were reviewed from 1952-1982. There were 37 cases of adenoid cystic carcinoma of the breast (ACC) from a total of 40,350 invasive breast tumors. Patient survival, complications, and pathologic sections were reviewed. Only 14 of 27 surgical pathology slides available for review could be confirmed histologically as ACC. All patients were white females with a mean age of 64 years. The tumor remained localized to the breast in all cases. Nine patients had either radical or modified radical mastectomy, four patients had either simple mastectomy or lumpectomy, and one patient refused treatment. There was no evidence of axillary node involvement, metastases, or local recurrence after excision. At the time of follow-up, nine patients were alive and disease free and four died of disease unrelated to their breast cancer. The one patient who died of breast cancer had a radical mastectomy and survived 11.7 years after diagnosis. It is concluded that ACC has a favorable biologic behavior characterized by a prolonged clinical course and good prognosis. Simple mastectomy is all that is required as initial treatment, and a chest x-ray and thorough physical examination looking for local recurrence is all that is needed for follow-up. PMID- 3030201 TI - Cytomegalovirus as a risk factor in living-related renal transplantation. A prospective study. AB - Forty-four living-related donor kidney (LRD) recipients (19 HLA-identical and 25 haploidentical) were followed prospectively to determine the posttransplant incidence and sequelae of cytomegalovirus (CMV) infection as they relate to the CMV status of recipients and donors. CMV titers were measured in all patients before transplantation by an immunofluorescent assay (IFA). Recipients similarly had CMV titers measured at selected intervals after transplant and during febrile episodes. Appropriate viral cultures were simultaneously performed. Laboratory evidence of infection was correlated with symptoms and signs of active CMV disease. Mean follow-up period was 20 +/- 12 months with a range of 3-51 months. Three patients were excluded due to early acute rejection resulting in graft loss. Twenty-eight of 41 donors (68%) and 22 of 41 recipients (54%) had positive CMV titers before transplantation. Six of 41 recipients (15%) subsequently developed clinical and laboratory evidence of CMV infection: three of 19 seronegative recipients and three of 22 seropositive recipients. All six patients received kidneys from seropositive donors. Four patients had severe CMV disease (2 seronegative, 2 seropositive), whereas two patients had leukopenia and fever only. Two patients with severe CMV infections subsequently lost their grafts due to unrelated causes. Overall, actual patient and graft survival of the entire group is 95% and 82%, respectively. In conclusion, individuals who receive LRD kidneys from seronegative individuals are unlikely to develop CMV infection, and transplantation of seropositive LRD kidneys may be associated with transmission of CMV in susceptible recipients regardless of their serologic status. With appropriate management of CMV illness in the posttransplant period, LRD kidney donation is safe and efficacious and should not be discouraged on the basis of pretransplant CMV serology in any donor-recipient pairing. PMID- 3030202 TI - Azido cAMP, [32P]8N3 cAMP: a probe to study epididymal sperm maturation. PMID- 3030203 TI - Impaired adaptive receptor regulation: an index of aging? AB - Many neuroendocrine functions are altered in old animals and their study may represent important steps in the understanding of the mechanisms of aging. A deeper insight, however, can be achieved by investigating the responsiveness to stimuli, which may reveal alterations not evident in the unstimulated conditions. At this level of study, many of such impairments have been found to be caused by receptor changes. In the present paper a third level of study is suggested in order to gain evidence of some remote failure of adaptive processes strictly linked to intimate mechanisms of aging. As at the second level of study different receptor characteristics can frequently be found at the basis of age-related alterations of biological responsiveness, at the proposed third level altered capacity of receptor regulation may be hypothesized as responsible for altered cell adaptation following hormone and drug stimuli. Experimental data are given which support this view. The possibility that receptor regulation may be used as an index of aging is suggested. This hypothesis leads to the problem of judging the validity of biological parameters deputed to represent good indices of aging. In order to solve this problem, the potential use of a mathematical model of mortality kinetics is discussed. PMID- 3030204 TI - In vitro vasorelaxing activity of suloctidil. AB - The vasorelaxing effect of suloctidil was evaluated in isolated rat and rabbit aorta and in isolated rabbit mesenteric and saphenous artery. Suloctidil inhibited contractions induced by increasing extracellular calcium in depolarized arteries, mainly in a competitive way. In the rat aorta, the pA2 value was 7.50 for suloctidil, while pA2 values of 9.96, 7.90 and 8.10 were obtained for nifedipine, cinnarizine and verapamil, respectively. Suloctidil more potently inhibited calcium-induced contractions in small arteries (mesenteric and saphenous artery), than in the aorta. Suloctidil also reduced the tonic component of the responses to norepinephrine. In contrast to the effects on calcium-induced contractions, the effects of suloctidil on norepinephrine-induced responses was mainly noncompetitive. In addition, and unlike cinnarizine and verapamil, high concentration of suloctidil also reduced the phasic component of contractile responses to norepinephrine. Furthermore, unlike nifedipine, verapamil, diltiazem and cinnarizine, suloctidil was devoid of a negative inotropic effect in spontaneously beating guinea-pig atria. In conclusion, suloctidil behaves as a Ca2+-channel blocker in arteries and displays an additional mode of action that could include receptor-operated Ca2+-channels or an intracellular site of action. In addition, suloctidil was found to affect small arteries more than the aorta, and not to affect the atria. PMID- 3030205 TI - Myocardial and vascular effects of perindopril, a new converting enzyme inhibitor, during hypertension development in spontaneously hypertensive rats. AB - The effects of a new angiotensin I converting enzyme inhibitor (ACEI), perindopril (P), on genetic hypertension development (GHD), on mesenteric arteriolar compliance and reactivity to noradrenaline and on myocardial and aortic hypertrophy, have been investigated at intervals in SHRs. P (4 mg/kg, q.d.) was administered by gavage from the 4th to the 20th week of age and measurements were performed 20 hr after the preceding drug intake. P completely prevented GHD, maintaining systolic blood pressure (SBP) below 130 mmHg during the whole treatment period. Furthermore, 7 weeks after treatment withdrawal, SBP of previously treated SHRs was still significantly lower than that of controls. Vascular compliance and internal diameter of the mesenteric arteriole were significantly increased in P-treated SHRs as compared to controls at the age of 20 weeks but these effects did not persist 7 weeks after treatment withdrawal. P did not affect mesenteric arteriolar reactivity to noradrenaline. Finally, P significantly reduced myocardial and aortic hypertrophy in SHRs. Thus P strongly opposed the morphological and functional vascular alterations which usually occur in SHRs during GHD. This property, presumably related to converting enzyme inhibition within the vascular wall, probably contributes to a large extent to, but is not exclusively responsible for, the drug-induced prevention of GHD. PMID- 3030206 TI - Effect of triflusal and other salicylic acid derivatives on cyclic AMP levels in rat platelets. AB - The effect of triflusal, acetylsalicylic acid (ASA), and of their principal metabolites 2-hydroxy-4-trifluoromethylbenzoic acid (HTB) and salicylic acid (SA), alone or combined with dypiridamole (DIP) and/or PGE1 on cyclic AMP levels in washed rat platelets (37 degrees C, 4 min), has been determined. DIP at 0.1 mM increased cyclic AMP levels by 25%. The effect of triflusal and HTB was significant at therapeutic concentrations of triflusal (1 mM: 36% increase) and HTB (0.5 mM: 37% increase). The effect of HTB was always greater than that of triflusal. ASA, at 1 mM and 5 mM, alone or combined with PGE1 was without effect. When 1 mM triflusal was combined with 0.1 mM DIP an increased effect was obtained (95%). ASA, at the highest concentration tested (5 mM), did not modify the DIP induced increase of cyclic AMP levels. PMID- 3030208 TI - Further evidence for the involvement of cyclic AMP in Ca2+-dependent, but not Ca2+-independent, noradrenaline release in the rabbit pulmonary artery. AB - Strips of the rabbit pulmonary artery were incubated with 3H-noradrenaline and subsequently superfused. In strips superfused with Ca2+-free solution containing K+ 64.7 mmol/l, tritium overflow was stimulated by introduction of Ca2+ 1.6 mmol/l; in strips superfused with physiological salt solution, stimulation was carried out with veratridine 30 mumol/l, acetylcholine 1 mmol/l (in the presence of atropine 1 mumol/l) or tyramine 100 mumol/l. The Ca2+-, veratridine- or acetylcholine-induced tritium overflow was facilitated by forskolin, AH 21-132 (a cAMP phosphodiesterase inhibitor) and/or 8-Br-cAMP, i.e. compounds which increase intraneuronal cAMP content. In contrast, the Ca2+-independent tyramine-evoked tritium overflow was not affected by these drugs. It is concluded that cAMP is involved in the regulation of stimulation-evoked Ca2+-dependent noradrenaline release, and that the sympathetic nerve terminals are endowed with an adenylate cyclase system. Facilitation of release may be caused by an alteration of Ca2+ influx or utilization. PMID- 3030207 TI - Effect of the amino-oxazoline derivative S-3341 on pre- and postjunctional alpha adrenoceptors in isolated blood vessels. AB - In segments of isolated dog saphenous veins, S-3341 (10(-8) to 10(-4) M) induced concentration-dependent contractions which, like those to clonidine, were progressively inhibited by increasing concentrations of yohimbine (10(-8) to 10( 6) M) but not by prazosin (10(-8) to 10(-6) M). In strips of the rabbit main pulmonary artery, previously labelled with [3H]noradrenaline and mounted for superfusion, S-3341 and clonidine both decreased the overflow of [3H] caused by electrical stimulation; these effects of S-3341 and clonidine were inhibited by 10(-6) M of yohimbine but not by 10(-6) M of prazosin. Our results illustrate that both S-3341 and clonidine cause contraction by activation of postjunctional alpha 2-adrenoceptors and decrease noradrenaline-release by stimulating prejunctional alpha 2-adrenoceptors. At the postjunctional alpha 2-adrenoceptors clonidine is 27 times more potent than S-3341, while at the prejunctional alpha 2 adrenoceptors, clonidine is only 15 times more potent than S-3341. PMID- 3030209 TI - Delta receptors in the rat vas deferens. AB - The effects of the delta-selective antagonist ICI 174864 and naltrexone on the dose-response curves to the mu-selective agonist RX 783006 and D-ala-D-leucine enkephalin (DADL) have been investigated in the rat isolated vas deferens preparation (RVD) set up in Krebs solution containing half the normal Ca++ concentration. The results obtained provide very strong evidence for the existence of both mu- and delta-receptors in this preparation. The Ke values of a number of opioid antagonists v. DADL in the RVD have been determined and compared to the Ke values obtained in the mouse vas deferens assay (MVD). The very good correlation (slope = 1.01, r = 0.98) obtained between the Ke values from the 2 preparations confirms the presence of a delta-receptor in the RVD. PMID- 3030210 TI - Prevention by nimodipine, a calcium entry blocker, of the effect of alpha 2 adrenoceptor blocking agents on noradrenaline release: differential effects of nimodipine, on [3H]noradrenaline and [14C]acetylcholine release measured concomitantly from the guinea-pig ileum. AB - The calcium channel blocker, nimodipine (Bay e 9736), inhibited the evoked (1 Hz, 1 msec, 180 pulses) release of [14C]acetylcholine ([14C]ACh) while it enhanced the release of [3H]noradrenaline ([3H]NA) measured concomitantly from the same longitudinal muscle strip of guinea-pig ileum loaded with [14C]choline and [3H]noradrenaline. These effects of nimodipine were concentration-dependent: in 0.1 microM concentration yielding maximal inhibition of [14C]ACh release and in 0.5 microM concentration yielding half maximal stimulation of [3H]NA-release. Enhancement of [3H]NA-release by nimodipine was strongly dependent on stimulation frequency while the inhibitory effect of nimodipine on release of [14C]ACh did not depend on stimulation frequency in the 0.5-10 Hz range. In addition, in the presence of nimodipine when the release of [3H]NA was enhanced, i.e. when a significant negative feedback operation could have been expected, alpha 2 adrenoceptor blocking agents (yohimbine and idazoxane) failed to enhance [3H]NA release. It is proposed that nimodipine somehow exerts a presynaptic alpha 2 adrenoceptor blocking property at noradrenergic axon terminals through an effect on calcium entry, which may be an essential step in events linking alpha 2 adrenoceptor activation and inhibition of noradrenaline release. PMID- 3030211 TI - Further evidence for tonically functioning presynaptic beta-adrenoceptors in guinea-pig pulmonary arteries. AB - Effects of dl- and d-carteolol alone on the release of tritium evoked by transmural field stimulation at 1 Hz were investigated in spirally cut guinea-pig pulmonary arteries preloaded with [3H]-norepinephrine. Cumulatively applied dl carteolol 10(-8) M, 10(-7) M and 10(-6) M readily inhibited the release of tritium in a concentration-dependent manner, whereas the same concentrations of d carteolol produced no effect. In conclusion, presynaptic beta-adrenoceptors in guinea-pig pulmonary arteries tonically function to facilitate the release of the transmitter norepinephrine. PMID- 3030212 TI - Thoracic computed tomography in the preoperative evaluation of primary bronchogenic carcinoma. AB - One hundred seventy-four patients with bronchogenic carcinoma underwent computed tomography (CT) as part of their preoperative evaluation. Overall, CT had a sensitivity of 86%, a specificity of 78%, and an accuracy of 81% in identifying mediastinal lymph node metastases. In patients with a central tumor, the sensitivity was 93%, the specificity 74%, and the accuracy 83%. In patients with a peripheral tumor, the respective percentages were 55%, 82%, and 77%. Only 11 of 66 patients with a peripheral tumor had mediastinal metastases, and five of these patients had a normal CT scan. Conversely, 43 of 64 patients with a central tumor and mediastinal lymph node enlargement on the CT scan had unresectable disease, compared with only one of 44 patients without such enlargement. We conclude that CT is not useful in the evaluation of patients with a peripheral tumor; however, it is useful in determining which patients with a central tumor do not require a surgical staging procedure prior to thoracotomy. PMID- 3030213 TI - Ectopic thyroid tissue on thallium/technetium parathyroid scan. AB - The thallium 201/technetium 99m pertechnetate radionuclide study is becoming widely accepted as a means of localizing abnormal or aberrantly located parathyroid tissue. We describe a case in which ectopic retrosternal thyroid tissue appeared as a parathyroid adenoma on thallium-201/technetium-99m pertechnetate scan. Physicians who use this radionuclide study should be aware of the possibility of false-positive images within the mediastinum. PMID- 3030214 TI - Gastric lymphoma associated with human T-cell leukemia virus type I. AB - A 41-year-old woman presented with a gastric lymphoma. A total gastrectomy was performed, and the tumor was found to consist of T cells of the helper/inducer (E+, Leu-1+, Leu-2a-, Leu-3a+) phenotype. The patient was seropositive for T-cell leukemia virus type I, and the tumor cells contained the proviral genome. PMID- 3030215 TI - Cytomegalovirus mononucleosis-associated antral gastritis simulating malignancy. AB - A 44-year-old man with epigastric pain, fever, and atypical lymphocytosis was found to have severe antral gastritis with radiologic and endoscopic features suggestive of malignancy. Histopathologic and serologic findings were diagnostic of cytomegalovirus gastric infection. Clinical involvement of the stomach in the course of cytomegalovirus mononucleosis is unique, and this case expands further the spectrum of clinical disease in the otherwise healthy adult. PMID- 3030216 TI - [Isolated replacement of the tricuspid valve with a bioprosthesis. Apropos of 22 cases in Abidjan]. AB - The authors reviewed the operative and function results of 24 isolated tricuspid valve replacements with bioprostheses in 22 patients. The patient population was young (average age 15 +/- 8 years). The surgical indication was massive tricuspid regurgitation due to chronic parietal endocarditis in 19 cases and to bacterial endocarditis in 3 cases. Tricuspid valve replacement was associated with 19 right ventricular endocardectomies, 2 direct closures of ventricular septal defects, 2 Wooler mitral valvuloplasties and 1 pericardectomy. The operative mortality was 13.5% and the secondary mortality 13.5%. Of the 16 survivors, 13 are in the NYHA Class I with no regular medical therapy. Their cardiothoracic ratio has slightly decreased. Two patients have permanent atrial fibrillation, and 12 have acquired definitive complete right bundle branch block. Eight of these patients had significant improvement of atrial and right ventricular pressures, of Yu's index and cardiac index at postoperative catheterisation. Three of the 16 patients developed progressive calcific degeneration of their bioprostheses. They are among the 6 patients who have been followed up for more than 3 years. There was no mortality at reoperation. Isolated tricuspid valve replacement by bioprosthesis was chosen despite the young age of these patients because of the disadvantages of mechanical prostheses which are associated with a much higher mortality related to incarceration and thrombosis of the prosthesis. The relatively high operative and secondary mortality in this series of isolated tricuspid valve replacement compared to mitral, aortic or micro-aortic valve replacement, is related to the gravity of the underlying causal pathology. PMID- 3030217 TI - [Estradiol-17beta, testosterone, progesterone and cortisol levels in pregnant cows after ACTH administration in various stages of pregnancy]. PMID- 3030219 TI - Antibodies to MuMTV proteins in the sera of mammary carcinoma patients' healthy daughters. AB - The presence of antibodies to MuMTV-related proteins in sera of 31 mammary carcinoma patients' healthy daughters have been tested by indirect immunoperoxidase method (ELISA). Ten of them (32.3 +/- 8.4%) reacted with MuMTV proteins preparations, but not with MuLV, RaLV, SSV, MPMV proteins or preparations of mouse embryonic tissues. The percent of positive sera in this group is almost ten times higher than among women without breast cancer in family history (earlier published data). By immunoblotting method the specificity for five positive sera was determined. Two sera react with p27, one with p27 and to a less extent with gp52, one-only with gp52 and one more serum with both proteins equally. Mothers of the daughters with positive-reacted sera were characterized by premenopausal tumors, detected on early (I, IIa) clinical stage of disease. PMID- 3030218 TI - Enhanced adrenal sensitivity to exogenous cosyntropin (ACTH alpha 1-24) stimulation in major depression. Relationship to dexamethasone suppression test results. AB - ACTH alpha 1-24 (cosyntropin) (250 micrograms by intravenous bolus) was given to 38 medicated patients with major depressive disorder (MDD) and to 34 normal control subjects. Patients with MDD had significantly higher plasma cortisol concentrations and significantly higher increases in plasma cortisol levels 60 minutes after cosyntropin infusion than did control subjects. Patients who were nonsuppressors in the dexamethasone suppression test had significantly higher 60 minute cortisol concentrations and cortisol increases than did normal subjects and patients with MDD who were suppressors. There were significant, strongly positive correlations between cortisol secretory responses to cosyntropin and postdexamethasone cortisol concentrations in patients with MDD. These findings confirm that adrenal sensitivity to corticotropin (ACTH) is enhanced in MDD and suggest that this endocrine abnormality may be related pathophysiologically to the resistance of cortisol secretion to dexamethasone suppression. PMID- 3030220 TI - [Use of CT in the staging and follow-up of small-cell bronchial cancer]. AB - In 54 patients with small-cell bronchial carcinoma, investigated with conventional X-ray technique and computer tomography, CT afforded a considerable extension of findings in regard to mediastinal tumor and metastatic spread, as well as in the detection of supraclavicular and pleural metastases. Also, the process of remission and renewed tumor growth could be observed better. Thus, CT constitutes quite an acquisition to therapy planning. The investigations carried out simultaneously in the brain region did not appreciably add to the findings in comparison to scintigraphy. In contrast, in the abdominal region, we have discovered liver metastases, subrenal metastases and lymph-node metastases both at first investigations and at controls. At post-mortem, however, we found kidney metastases that were not described in the CT but which may possibly have arisen prefinally. Altogether, thoracal and abdominal investigations with CT prior to onset of therapy and subsequent control are a valuable addition to therapy planning. PMID- 3030221 TI - Cytomembranous inclusions observed in acquired immunodeficiency syndrome. Clinical and experimental review. AB - Two types of cytomembranous inclusions commonly occur in patients with acquired immunodeficiency syndrome (AIDS) and AIDS-related conditions: tubuloreticular inclusions (TRI) and cylindrical confronting cisternae (CCC). CLinical and experimental studies both indicate that the occurrence of TRI in AIDS, systemic lupus erythematosus, or viral infections is directly related to endogenous systemic or local elevations of type I interferons (interferons alpha or beta). Moreover, these inclusions develop in patients or rhesus monkeys undergoing interferon alpha treatment. Rarely had CCC been reported in human tissue before the AIDS epidemic, but they were well known to develop in the hepatocytes of chimpanzees after experimental inoculation with human non-A, non-B hepatitis agent. In AIDS, CCC frequently coexist with TRI, and their appearance is associated with disease progression. A relationship of CCC to type I interferons is indicated by recent in vitro investigations, but clinical discordances in their appearance of TRI, and CCC suggest some differences in their pathogenesis, possibly related to cellular susceptibility or etiologic cofactors. PMID- 3030222 TI - Comparison of cytomegalovirus infection of brain and lung in a patient with subacute encephalopathy of acquired immunodeficiency syndrome. AB - The ultrastructure of cytomegalovirus infection of glial cells in an area of demyelination in the brain of a patient with subacute encephalopathy associated with acquired immunodeficiency syndrome is described. By comparison with the lung of the same patient, the cytomegalovirus-infected brain showed a lower density of complete virions per cell and a higher proportion of defective viral particles. PMID- 3030223 TI - The effectiveness of bronchoscopy in the diagnosis of Pneumocystis carinii and cytomegalovirus pulmonary infections in acquired immunodeficiency syndrome. AB - We evaluated the sensitivity of bronchoscopy for the diagnosis of Pneumocystis carinii and cytomegalovirus pulmonary infections in patients with acquired immunodeficiency syndrome. The antemortem and postmortem diagnoses were compared in 36 patients who underwent fiberoptic bronchoscopy within two weeks of death. In autopsy-proved cases of Pneumocystis carinii pneumonia (PCP), the organism was correctly identified antemortem in 22 (88%) of 25 cases, including 94% of adequate transbronchial bronchoscopic biopsy specimens, 95% and 88% of bronchoalveolar lavage (BAL) cell blocks and smears, respectively, and 79% of brushing. In 11 patients who underwent simultaneous adequate biopsy, BAL, and brushings, the diagnostic sensitivity for PCP was 100%. The negative predictive value of bronchoscopy for PCP was 85%. Bronchoscopy yielded the diagnosis of cytomegalovirus infection in only 55% of autopsy-proved cases. Diagnostic sensitivity was also reduced when an important diagnostic procedure, such as transbronchial biopsy or BAL, was inadequate or not performed. PMID- 3030224 TI - Simultaneous cytomegalovirus and herpes simplex virus pneumonia. AB - A 35-year-old patient who underwent renal transplant developed persistent fever, hypoxemia, and diffuse interstitial pulmonary infiltrates one month after allograft implantation. An open lung biopsy specimen demonstrated simultaneous infection with cytomegalovirus and herpes simplex virus type 1. This initially was unsuspected on routine histology, but was confirmed by the demonstration of both viruses with immunofluorescence, as well as the timely recovery in culture of both. The clinical and pathologic implications of an accurate diagnosis of such a simultaneous infection are discussed. PMID- 3030225 TI - Fatal case of Epstein-Barr virus-induced lymphoproliferative disorder associated with a human immunodeficiency virus infection. AB - This is a case report of an Epstein-Barr virus-induced polyclonal lymphoproliferative disorder in a presumably immunocompromised patient with Western blot-confirmed antibodies to human immunodeficiency virus. Postmortem examination revealed a diffuse lymphoplasmacytic infiltrate with prominent numbers of immunoblasts involving multiple organs and resulting in multiple organ system failure. PMID- 3030226 TI - 'Primitive neuroectodermal tumors'. PMID- 3030227 TI - A method for cytologic examination of cartilaginous lesions. AB - Chondrocytes, dissociated from their matrix with trypsin and clostridial collagenase, retain their cytologic integrity. Successful preparations have been made from postmortem as well as surgical specimens. The method may lend itself to diagnosis of both neoplastic and developmental lesions. PMID- 3030228 TI - Fine-needle aspiration of salivary gland lesions. Comparison with frozen sections and histologic findings. AB - The results of 171 salivary gland fine-needle aspirates, with subsequent histologic correlation, were compared with those from previous head and neck series and analyzed for diagnostic accuracy. Cytologically, 118 cases were diagnosed as benign; 51, malignant; and two, insufficient for diagnosis. The false-negative rate was 4.7%, and the false-positive rate was 3.5%. Pleomorphic adenoma, mucoepidermoid carcinoma, chronic sialadenitis, and malignant lymphoma were the lesions most frequently misdiagnosed. Corresponding frozen sections (available in 38 cases) showed an exact correlation with the final surgical pathologic diagnosis in 58% of the cases, with no false-positive diagnoses but an 11% false-negative rate. PMID- 3030229 TI - Two chromosomally different cell populations in a partly cellular congenital mesoblastic nephroma. AB - The conventional fibromatous and cellular areas of a congenital mesoblastic nephroma were studied using electron microscopy, and in vitro and cytogenetic methods. In light- and electron-microscopic studies, as well as in cell cultures, mature and immature mesenchymal cell types that corresponded to the fibromatous and cellular areas of the tumor were found. The conventional fibromatous portion of the tumor showed a normal chromosomal pattern, while the cellular, pleomorphic tumor area was characterized by an aneuploid clone with 54 chromosomes. The value of cytogenetic analysis of congenital mesenchymal renal tumors as a possible diagnostic tool in histologically questionable cases is discussed. PMID- 3030230 TI - Malignant mixed mullerian tumor of the fallopian tube. AB - Malignant mixed mullerian tumor of the fallopian tube is an extremely rare neoplasm, with less than 30 cases reported. Clinical signs and symptoms are usually nonspecific and include lower abdominal pain, fever, and vaginal bleeding. The age at diagnosis varies from 14 to 76 years, with a mean of 55 years. Prognosis is poor, with only one woman surviving more than five years. Surgery is the treatment of choice, although chemotherapy may be of some benefit. PMID- 3030231 TI - [Extra-articular synovial chondromatosis of the hand]. PMID- 3030233 TI - [Surgical treatment in a case of severe rachitic deformity of the lower extremities]. PMID- 3030232 TI - [Idiopathic hypophosphatemic osteomalacia arising in an adult: a case report]. PMID- 3030234 TI - Hepatotoxicity of the synthetic oestrogen hexoestrol in the female rat. AB - The synthetic oestrogen hexoestol is hepatotoxic to the female rat. Twenty-five animals were treated daily with hexoestrol at a dose of 60 mg/kg. Twenty-five more acted as controls. Five animals from each group were killed after 4, 12, 20, 30 and 40 days of treatment to permit serial evaluations of liver histopathology to complement serum biochemical investigations. There were no mortalities during the study and only modest external signs of toxicity. All the treated rats showed reductions in body weight and appetite and gains in liver weight. This latter effect was due to both cellular hypertrophy and hyperplasia. The principle finding of the liver histopathology was fatty change. This affected scattered individual cells, mainly in the midzonal region. All of the serum parameters of toxicity--which consisted of bilirubin, total protein, albumin, glycoprotein, alpha 2-macrofoetoprotein, and alkaline phosphatase--indicated liver impairment. Most of these also showed time-related changes consistent with an initial phase up to Day 20 of marked liver dysfunction superseded by a more adaptive phase thereafter. The hepatotoxicity described here seems sufficient to explain blood clotting defects observed elsewhere in oestrogen-treated rats. PMID- 3030236 TI - Herpesvirus simiae (B virus): replication of the virus and identification of viral polypeptides in infected cells. AB - The events and products of replication of Herpesvirus simiae (B virus) in Vero cells were studied. The time course of the synthetic events of DNA replication and protein synthesis were found to be similar to the processes of the herpes simplex viruses and SA 8. Infectious progeny virus were detected by 4 hours post infection and were first found extracellularly between 6 and 8 hours post infection (PI). As in the case of SA 8, all cell lines tested were permissive for lytic infection by B virus. Analyses of B virus-infected cells by SDS polyacrylamide gel electrophoresis (SDS-PAGE) revealed approximately 50 infected cell polypeptides (ICP) ranging in molecular weight from about 26,000 to 239,000 daltons. The kinetics of synthesis of the ICPs were also identified. At least nine glucosamine-containing glycopeptides were noted ranging from 133,000 to 29,000 daltons. PMID- 3030235 TI - Viruses as toxins. With special reference to paramyxoviruses. Brief review. PMID- 3030237 TI - Non-neutralizing monoclonal antibodies to a trypsin-sensitive site on the major glycoprotein of rotavirus which discriminate between virus serotypes. AB - Monoclonal antibodies were derived to a human rotavirus purified from stools. Three of the antibodies immunoprecipitated the rotavirus outer capsid glycoprotein gp 34 and were non-neutralizing. These antibodies reacted by enzyme immunoassay with cultivable rotaviruses showing the "long" RNA electropherotype but were inefficient as detectors of "long" RNA pattern rotaviruses in stools. Treatment of SA 11 rotavirus with 7.5 micrograms/ml porcine trypsin for 30 minutes at 37 degrees C irreversibly reduced binding of the antibodies to SA 11 rotavirus in enzyme immunoassays by 50 per cent. Binding was abolished in the presence of rotavirus-negative faecal extracts. These results indicate that non neutralizing sites on gp 34 of rotaviruses can vary with RNA electropherotype and serotype, and that levels of trypsin currently in use to assist growth of rotaviruses in cell culture may alter the serological profile of the viruses. PMID- 3030239 TI - DMSO induces reactivation of cytomegalovirus in vitro from spleens of latently infected mice. Brief report. AB - Reactivation of murine cytomegalovirus in spleen explant cultures from latently infected CD 1 mice was studied. A significant increase in the incidence of reactivation was observed when 200 mM DMSO was included in the culture medium. Virus could also be reactivated from the bone marrow of some mice. PMID- 3030240 TI - VSV binding to lipids from different cell lines. Brief report. AB - Gangliosides from different cells were tested for their inhibiting activity on VSV attachment to CER cells and goose erythrocytes. This inhibiting activity was enhanced when gangliosides were inserted into liposomes, in particular if containing phosphatidylserine. PMID- 3030238 TI - Serologic differences among nondefective reticuloendotheliosis viruses. AB - Antigenic relationships among 26 isolates of reticuloendotheliosis virus (REV) obtained from several avian species were compared by cross neutralization tests with polyclonal chicken sera and by immunofluorescent assays with monoclonal antibodies to REV strain T. The isolates were all strongly related by neutralization assays and thus probably constitute a single serotype. However, 3 antigenic subtypes were suggested by minor but distinct differences in neutralization titers. The validity of these 3 subtype designations was confirmed by differential reactivity of viral isolates to selected monoclonal antibodies. Subtype-associated differences in serum antibody titers were noted following the inoculation of chickens with the REV isolates. PMID- 3030242 TI - [Infection and asthma. Part 4. Viral studies by serological technics on infants and children with either asthma, wheezing, or prolonged cough]. PMID- 3030241 TI - Genome isomerism in two alphaherpesviruses: Herpesvirus saimiri-1 (Herpesvirus tamarinus) and avian infectious laryngotracheitis virus. Brief report. AB - Genome isomerism of avian infectious laryngotracheitis virus (ILT) and Herpesvirus saimiri-1 (HVS-1) (formerly H. tamarinus) was investigated using restriction endonuclease analysis and scanning densitometry. ILT DNA was found to have 2 isomers with a molecular weight of 109 X 10(6). HVS-1 DNA was found to have four isomers with a molecular weight of 102 X 10(6). PMID- 3030243 TI - [Current status of the study of myocarditis and endomyocardial biopsy]. PMID- 3030244 TI - Angiotensin-converting enzyme in neurosarcoidosis. PMID- 3030245 TI - An alert for motor neuron diseases and peripheral neuropathy: monoclonal paraproteinemia may be missed by routine electrophoresis. PMID- 3030246 TI - Sporadic non-A, non-B hepatitis and Epstein-Barr hepatitis associated with the Guillain-Barre syndrome. AB - Three cases of Guillain-Barre syndrome (GBS) complicating sporadic non-A, non-B viral hepatitis (NANB) are reported. One case occurred in the incubation period for NANB hepatitis, and the other cases were noted to have hepatitis at the time of hospital admission due to paralysis. Three patients (13%) in a prospective series of 23 patients with GBS were found to have subclinical hepatitis (two patients with NANB hepatitis, and one patient with Epstein-Barr viral hepatitis). This unexpectedly high incidence of hepatitis associated with GBS is discussed and a possible causal link between NANB hepatitis and surgery-related GBS is proposed. PMID- 3030247 TI - [Oncology]. PMID- 3030248 TI - The effect of a fluorocarbon surfactant on the surface tension of the endodontic irrigant, sodium hypochlorite. A preliminary report. PMID- 3030249 TI - Experimental studies on mechanism of the Meniere's attack: investigation into vestibulo-cochlear response of the guinea pig induced by potassium ion. AB - After introducing potassium ion through the round window into the perilymphatic space of 40 guinea pigs by means of iontophoresis, physiological and histochemical investigations were performed to determine the role of the high perilymphatic potassium concentration in the vertiginous attack of Meniere's disease. About 15 min after the iontophoretic procedure, electronystagmography revealed irritative nystagmus for the first 5 min and then paralytic nystagmus for the following 6 to 24 hr. Histochemical analysis of the vestibular sensory epithelia revealed the increased activity of succinic dehydrogenase and Na-K ATPase during irritative nystagmus and the decreased activity during paralytic nystagmus. The Na-K-ATPase activity was dominant in the synaptic area between the hair cells and the nerve-endings of the vestibular sensory epithelia. There was some delay between the reversal of nystagmus-direction and the change of enzyme activity. This delay was thought to be produced by the central regulatory mechanism for the disturbed tonus-balance in the vestibular nucleus. On the other hand, electrocochleography revealed the decrease of the action potential without any initial irritative cochlear sign, and the enzyme activity of the cochlear sensory cells was decreased from the beginning. PMID- 3030250 TI - Paraneoplastic optic neuritis and external ophthalmoplegia. AB - A 58 year old man developed bilateral optic neuritis and external ophthalmoplegia nine months before presentation with a small cell carcinoma of the lung. There was no evidence of central nervous system metastases and his ocular symptoms responded to corticosteroid therapy. PMID- 3030251 TI - Restriction endonuclease cleavage analysis of DNA from two bovine herpes mammillitis viruses, isolated in different parts of Australia. PMID- 3030252 TI - The effect of pasteurisation on bovine leucosis virus-infected milk. PMID- 3030253 TI - Bovine leucosis virus: antibody in cattle in New South Wales. PMID- 3030254 TI - Hepatic inclusion bodies in the liver of a koala (Phascolarctos cinereus). PMID- 3030255 TI - Antenatal screening for congenital infection with rubella, cytomegalovirus and toxoplasma. AB - The sera of 3,463 pregnant women were screened, at the first antenatal visit, for antibodies to rubella, cytomegalovirus (CMV) and Toxoplasma gondii. Rubella antibodies were detected in 97.5%, CMV antibodies in 71% and toxoplasma antibodies in 45% of women. Asymptomatic toxoplasmosis occurred during pregnancy in 3 of 609 (0.5%) and primary CMV infection in 5 of 338 (1.5%) initially seronegative women whose sera were retested at the end of their pregnancies. The observed incidence of toxoplasmosis was similar to that calculated on the basis of the age-related prevalence of antibodies found in this study. On the basis of these observations it is estimated that congenital toxoplasmosis and congenital CMV infection due to primary maternal infection each occurs in up to 2/1,000 infants in this community. Very few of these infants have obvious abnormalities at birth, but follow-up studies elsewhere have shown that many of them suffer significant long-term sequelae. Routine antenatal screening for rubella antibodies is well established and justifiable for this preventable congenital infection. However, routine antenatal screening for CMV antibodies cannot be justified at present, since neither immunization nor treatment is available. Further information is required to determine the cost-effectiveness of routine antenatal screening for toxoplasma antibodies and treatment of proven maternal infection during pregnancy. PMID- 3030256 TI - Emergency serum and urine HCG analyses with the 'Tandem ICON' procedure. AB - A comparison between new urine and serum enzyme immunoassay techniques and existing serum radioimmunoassay techniques for the detection of HCG in the diagnosis of ectopic and early intrauterine pregnancy was undertaken. Urine HCG estimations by enzyme immunoassay were not found to be adequate for the exclusion of ectopic pregnancy due to a false negative rate of 12.5% (2 of 16 patients). Serum HCG estimations by enzyme immunoassay were found to compare favourably with radioimmunoassay techniques in the detection of HCG in both ectopic and early intrauterine pregnancy. PMID- 3030257 TI - Flow cytometry of blastogenesis and the concomitant viability assay of lymphocytes stained with 4', 6 diamidino-2-phenylindole. AB - The blastogenic response of lymphocytes induced by concanavalin A (Con A), phytohemagglutinin (PHA), pokeweed mitogen (PWM) and by paraperiodic acid (H5IO6) was assayed by flow cytometry of 4',6 diamidino-2-phenylindole (DAPI) stained nuclei as well as by thymidine (3H-TdR) incorporation rates. The DNA content in DAPI-stained nuclei of viable and dead human, rat and mouse lymphocytes, was readily distinguishable from each other by flow cytometric methods (FCM). PMID- 3030258 TI - [Development and course of IBR antibody titers after vaccination with Bovigrip plus]. PMID- 3030259 TI - [Herpes problems from a comparative medical viewpoint]. PMID- 3030260 TI - [Incidence of serum neutralizing antibodies to various variant strains of the infectious bronchitis virus in chickens in South Wurttemberg]. PMID- 3030261 TI - [Suitability of the immunodiffusion test for determining humoral antibodies to bovine herpesvirus (BHV 1)]. PMID- 3030263 TI - Hormonal control of fructose 2,6-bisphosphate concentration in the HT29 human colon adenocarcinoma cell line. Alpha 2-adrenergic agonists counteract effect of vasoactive intestinal peptide. AB - Vasoactive intestinal peptide (VIP) was found to cause a dose-dependent decrease in fructose 2,6-bisphosphatase concomitant with an increase in cyclic AMP in cultured HT29 cancer cells from human colon. The maximum effect was a 41% decrease obtained with 10 nM-VIP, and half-maximum effect was obtained with 0.75 nM-VIP. The effect of 2.5 nM-VIP was almost totally counteracted (i.e. fructose 2,6-bisphosphate concentration was restored) by either adrenaline (1 microM) or the alpha 2-adrenergic agonist UK-14304 (1 microM); the alpha 2-agonist clonidine (1 microM) was less efficient, since the VIP effect was decreased by 72% only. The adrenaline effect was totally antagonized by 1 microM-yohimbine. It is concluded that, in the HT29 cancer cells, the fructose 2,6-bisphosphate-producing system is sensitive to variations of cyclic AMP concentration and is under the dual control of VIP and alpha 2-adrenergic receptors. PMID- 3030264 TI - Renal plasma membrane receptors for certain modified serum albumins. Evidence for participation of a heparin receptor. AB - Binding of formaldehyde-treated (f-alb), reduced-carboxymethylated (ac-alb) or reduced-acetamidated (am-alb) bovine serum albumins to purified rat renal plasma membranes was studied. Radioiodinated f-alb or ac-alb bound to kidney membranes while am-alb neither bound significantly nor competed with f-alb binding to kidney membranes. The binding was specific, saturable and heat- and proteinase sensitive. Competition studies showed that f-alb and ac-alb sites may be the same on these membranes. To determine the role played by charge in binding, competition experiments with polyanions were performed. Polyanions such as nucleic acid or glycosaminoglycans were effective competitors of f-alb binding to cell membranes. Heparin was especially inhibitory, being several-fold more so than chondroitin sulphate. Completely reduced and carboxymethylated albumin was a better competitor than its partially modified counterpart. Furthermore, f-alb was a significant competitor of [35S]heparin binding to kidney membranes. Also, partially purified heparin receptor demonstrated specific binding of 125I-f-alb. These data suggest that a heparin receptor is responsible for binding and internalization of intravenously injected f-alb. A Scatchard plot revealed two classes of receptors with dissociation constants of 3.2 X 10(-6) M and 4.7 X 10( 5) M. PMID- 3030262 TI - Epidermal growth factor mimics insulin effects in rat hepatocytes. AB - Epidermal growth factor (EGF) mimicked the effect of insulin to activate glycogen synthase and stimulate glycogen synthesis in isolated rat hepatocytes. Both agents required glucose (greater than 5 mM) and had similar time courses of action. The maximum effect of EGF was approx. 70% of that of insulin, and the half-maximally effective concentrations were 9 nM and 4 nM respectively. Combinations of the two agents produced additive responses. EGF also resembled insulin in its ability to inhibit the effects of 0.1-1.0 nM-glucagon on cyclic AMP and glycogen phosphorylase in hepatocytes. The maximum effect of EGF was approx. 70% of that of insulin, and the half-maximally effective concentrations were approx. 5 nM and 0.5 nM respectively. EGF and insulin inhibited phosphorylase activation by exogenous cyclic AMP, and inhibited cyclic AMP accumulation induced by forskolin. They also inhibited phosphorylase activation provoked by phenylephrine, but not by vasopressin. EGF added alone rapidly activated phosphorylase and increased cytosolic [Ca2+], but the effects were no longer apparent at 5 min and were smaller than those of vasopressin. Insulin did not induce these changes. In hepatocytes previously incubated with myo [3H]inositol, EGF did not significantly increase myo-inositol 1,4,5 trisphosphate. However, its ability to increase cytosolic [Ca2+] was blocked by neomycin, an inhibitor of phosphatidylinositol bisphosphate hydrolysis. It is concluded that some, but not all, of the effects of EGF in liver are strikingly similar to those exerted by insulin, suggesting that these agents may have some similar mechanisms of action in this tissue. PMID- 3030266 TI - 6-beta-Iodopenicillanate as a probe for the classification of beta-lactamases. AB - An inactivator of serine beta-lactamases, 6 beta-iodopenicillanate, can be utilized as a probe in the classification of beta-lactamases. It is a substrate for class-B Zn2+-containing beta-lactamase II. Although it inactivates enzymes from both classes A and C, it is much more efficient for the former group, with which it sometimes interacts following a branched pathway. On the basis of these observations, predictions are made concerning the class to which several enzymes belong. PMID- 3030265 TI - Guanine nucleotide binding protein involved in muscarinic responses in the pig coronary artery is insensitive to islet-activating protein. AB - In an attempt to identify the nature of guanine nucleotide binding protein(s) (G protein) involved in the acetylcholine (ACh)-induced (muscarinic) response of pig coronary-artery smooth muscle, we studied the effect of ADP-ribosylation of specific membrane protein(s) catalysed by islet-activating protein (IAP; pertussis toxin). The ACh-stimulated and guanine nucleotide-dependent activities of phosphatidylinositol 4,5-bisphosphate (PIP2) phosphodiesterase (PDE), assessed by the production of inositol 1,4,5-trisphosphate (IP3) from exogenously applied PIP2, were not modified, in either IAP-treated or non-treated cell homogenates used as the enzyme source. In intact tissues, pretreatment with up to 100 ng of IAP/ml inhibited neither the ACh-induced decrease in the amount of inositol phospholipids nor the increase in the amounts of phosphatidic acid and of inositol phosphates. IAP treatment increased the amount of cyclic AMP accumulated by isoprenaline. These observations suggest that G-protein which couples the muscarinic receptor to PIP2-PDE is insensitive to IAP. Such being the case, the nature of this protein(s) probably differs from that required for the regulation of adenylate cyclase activities (Ni or Gi). PMID- 3030268 TI - Inhibition by chlorogenic acid of haematin-catalysed retinoic acid 5,6 epoxidation. AB - Chlorogenic acid (3-O-caffeoylquinic acid) inhibited haematin- and haemoglobin catalysed retinoic acid 5,6-epoxidation. Some other phenol compounds (caffeic acid and 4-hydroxy-3-methoxybenzoic acid) also showed inhibitory effects on the haematin- and haemoglobin-catalysed epoxidation, but salicylic acid did not. Of the above compounds, caffeic acid and chlorogenic acid were potent inhibitors compared with the other two, suggesting that the o-hydroquinone moiety of chlorogenic acid and caffeic acid is essential to the inhibition of the epoxidation. Although caffeic acid inhibited retinoic acid 5,6-epoxidation requiring the consumption of O2, formation of retinoic acid radicals was not inhibited on the addition of caffeic acid to the incubation mixture. The above results suggest that caffeic acid does not inhibit the formation of retinoic acid radicals but does inhibit the step of conversion of retinoic acid radical into the 5,6-epoxide. PMID- 3030267 TI - Trypsin-induced increase in cyclic AMP concentration in rat thymocytes. An effect independent of calcium and calmodulin. AB - Trypsin produces a dose-related increase in cellular cyclic AMP concentration in rat thymocytes [Shneyour, Patt & Trainin (1976) J. Immunol. 117, 2143-2149; Segal & Ingbar (1983) Clin. Res. 31, 277A]. In the present study, I examined whether this effect of trypsin requires Ca2+ and whether it is modified by calmodulin. In fresh thymocytes suspended in standard medium (containing 1 mM-Ca2+), trypsin produced a concentration-dependent increase in cytoplasmic free Ca2+ concentration, which was evident at a concentration of 50 micrograms of trypsin/ml and reached maximal values at about 1 mg/ml. This effect of trypsin was very prompt in onset, almost immediate, and reached maximal values within 2-3 min. But in cells suspended in essentially Ca2+-free medium (6 nM free Ca2+), trypsin had no effect on cytoplasmic free Ca2+ concentration, which indicates that trypsin acted by increasing Ca2+ uptake rather than Ca2+ release from an intracellular pool. However, the increase in thymocyte cyclic AMP concentration produced by trypsin was independent of extracellular Ca2+ and was not influenced by calmodulin, because it was the same in the presence or absence of Ca2+ and was not changed by the calmodulin inhibitor trifluoperazine. I therefore suggest that in rat thymocytes the trypsin-induced increase in cyclic AMP concentration does not require Ca2+ and is not influenced by calmodulin. PMID- 3030269 TI - The development of cytochrome b-245 in maturing human macrophages. AB - Cytochrome b-245, the putative terminal component of the specialized cidal oxidase system of phagocytes, was measured in human monocytes in culture. There was a dramatic synthesis of the cytochrome, which increased by 27.3 +/- 2.0 pmol/day per 10(7) cells. This represents an increase of about 40%/day in the early stages and an overall 7-fold increase after 16 days. The protein content increased 3-fold over the same period, resulting in a doubling of the specific content of the cytochrome b. The newly synthesized cytochrome b was identified as that specifically located in the microbicidal oxidase electron-transport chain, as titration demonstrated that, at day 16 of maturation, 70% of the total membrane cytochrome b had a very low midpoint potential (-260 to -220 mV), characteristic of that found in this oxidase system. This cytochrome distributed with the plasma membrane on analytical subcellular fractionation, and a close relationship was observed between the maturation-induced increase in the concentration of this molecule and the capacity of the cells to produce superoxide. PMID- 3030270 TI - Five enzymes of the glycolytic pathway serve as substrates for purified epidermal growth-factor-receptor kinase. AB - Several enzymes of the glycolytic pathway are phosphorylated in vitro and in vivo by retroviral transforming protein kinases. These substrates include the enzymes phosphoglycerate mutase (PGM), enolase and lactate dehydrogenase (LDH). Here we show that purified EGF (epidermal growth factor)-receptor kinase phosphorylates the enzymes PGM and enolase and also the key regulatory enzymes of the glycolytic pathway, phosphofructokinase and glyceraldehyde-3-phosphate dehydrogenase (GAPDH), in an EGF-dependent manner. Stoichiometry of phosphate incorporation into GAPDH (calculated from native Mr) is the highest, reaching approximately 1. LDH and other enzymes of the glycolytic pathway are not phosphorylated by the purified EGF-receptor kinase. These enzymes are phosphorylated under native conditions, and the Km values of EGF-receptor kinase for their phosphorylation are close to the physiological concentrations of these enzymes in the cell. EGF stimulates the reaction by 2-5-fold by increasing the Vmax. without affecting the Km of this process. Phosphorylation is rapid at 22 degrees C and at higher temperatures. However, unlike the self-phosphorylation of EGF-receptor, which occurs at 4 degrees C, the glycolytic enzymes are poorly phosphorylated at this temperature. Some enzymes, in particular enolase, increase the receptor Km for ATP in the autophosphorylation process and thus may act as competitive inhibitors of EGF-receptor self-phosphorylation. On the basis of the Km values of EGF receptor for the substrate enzymes and for ATP in the phosphorylation reaction, these enzymes may also be substrates in vivo for the EGF-receptor kinase. PMID- 3030271 TI - Isolation and identification of uridine(5')-diphospho(1)-2,3-diacetamido-2,3 dideoxy-alpha-D- glucopyra nuronic acid from Pseudomonas aeruginosa P1-III. AB - A new uridine nucleotide was isolated from Pseudomonas aeruginosa P1-III (Habs serotype 5). On the basis of 13C-n.m.r. and p.m.r. spectroscopy, mass spectrometry, i.r.-absorption spectroscopy and circular dichrometry, the structure of the new compound was unequivocally identified as uridine(5') diphospho(1)-2,3-diacetamido-2,3-dideoxy-alpha-D-gl ucopyranuronic acid. PMID- 3030273 TI - Direct extraction and assay of bone tissue collagenase and its relation to parathyroid-hormone-induced bone resorption. AB - A method has been developed for the quantitative extraction of collagenase from as little as one 19-day-fetal-mouse calvarium. About 20-40 munits of collagenase are extracted per mg of tissue, all in a latent form that, after proper activation, shows the typical properties of mammalian collagenase. Culturing the calvaria for 2 days with parathyroid hormone (PTH) increases their procollagenase content up to 3-fold and induces bone resorption. Both PTH effects are prevented by cycloheximide, but not by indomethacin. Calcitonin inhibits resorption without affecting the PTH-induced procollagenase synthesis. The role of this synthesis is discussed in relation to the mechanisms of bone resorption. PMID- 3030272 TI - The ornithine requirement of urea synthesis. Formation of ornithine from glutamine in hepatocytes. AB - In hepatocytes, urea synthesis from glutamine is independent of added ornithine, even when rates are high after stimulation of glutamine metabolism by dibutyryl cyclic AMP, phenylephrine or vasopressin. Incubation with glutamine increases tissue [ornithine]. The increases parallel those of [N-acetylglutamate] under different conditions. The ornithine requirement of urea synthesis increases with increasing supply of ammonia. A function of the unique, highly regulated, glutaminase of liver may be to regulate ornithine synthesis. PMID- 3030274 TI - Competitive inhibition of human skin collagenase by N-benzyloxycarbonyl-L-prolyl L-alanyl-3-amino-2-oxopropyl-L-leucyl-L- alanylglycine ethyl ester. AB - A 'ketomethylene' peptide, N-benzyloxycarbonyl-L-prolyl-L-alanyl-3-amino- 2 oxopropyl-L-leucyl-L-alanylglycine ethyl ester, was synthesized and shown to be a fairly potent competitive inhibitor of human skin collagenase. PMID- 3030275 TI - Identification of ecdysone 22-long-chain fatty acyl esters in newly laid eggs of the cattle tick Boophilus microplus. AB - The five major apolar ecdysone esters present in newly laid eggs of the cattle tick Boophilus microplus have been purified by h.p.l.c. The quantities of the apolar esters present in the eggs were increased by administration of ecdysone to the mature females. G.c.-m.s. analysis, as their methyl esters, of the fatty acids released from the apolar ecdysone derivatives by alkali, coupled with positive-ion fast-atom-bombardment m.s. of the intact ecdysone esters, showed that the compounds consisted of a series of fatty acyl esters of ecdysone. The position of esterification of the ecdysone was established by p.m.r. spectroscopy. The combined data show that the novel apolar derivatives of ecdysone consist of the 22-palmitate, -palmitoleate, -stearate, -oleate, and linoleate esters respectively. Confirmation was obtained by comparison with synthetic ecdysone 22-palmitate. The significance of the ecdysone fatty acyl esters as a possible source of free hormone during embryogenesis is discussed. PMID- 3030276 TI - Chemical modification of the haem propionate of cytochrome c. A re-evaluation of the reaction of cytochrome c with a water-soluble carbodi-imide. AB - Horse heart and tuna heart cytochromes c were treated with the water-soluble carbodi-imide 1-(3-dimethylaminopropyl)-3-ethylcarbodi-imide. When the reaction is followed spectroscopically two kinetic phases are apparent. Alteration of the reactivity of the proteins with such ligands as CO, however, occurs in a single phase identical with the faster phase detected spectroscopically. The modified proteins both show spectroscopic and redox properties identical with those described for the tuna heart cytochrome c derivative by Timkovich [Biochem. J. (1980) 185, 47-57]. The use of radiolabelled carbodi-imide identifies two or three sites of reactivity. However, the addition of glycine methyl ester to the reaction mixture leads to the addition of nine glycine moieties in the case of the horse protein and seven in the case of the tuna protein, indicating a larger number of reactive sites than previously reported. A further set of reaction sites was identified by peptide mapping of the modified proteins, and these sites take part in intramolecular reactions leading to internal cross-linking and the formation of an enzymically indigestible 'core particle'. The haem group was identified as a site of reaction with the carbodi-imide, and is as a consequence covalently linked to the peptide by a bond in addition to the thioether bonds normally present. In the light of these findings, the alterations in the properties of the tuna protein, subsequent to reaction with the carbodi-imide, which have been previously explained in structural terms, must be re-evaluated. This study also highlights the importance of internal cross-link formation, which can occur by intramolecular nucleophilic attack, a process that has often been overlooked by investigators employing carbodi-imide modification of carboxylate groups in proteins. PMID- 3030278 TI - Purified rat lens aldose reductase. Polyol production in vitro and its inhibition by aldose reductase inhibitors. AB - The production of polyols in vitro by highly purified aldose reductase (EC 1.1.1.21) was monitored by g.l.c. In the presence of NADPH aldose reductase reduced glucose, galactose and xylose to the respective polyols sorbitol, galactitol and xylitol. The rates of formation of these polyols closely mirrored the Km values for the substrates obtained from kinetic measurements that monitored the rate of disappearance of NADPH. No polyol production occurred in the absence of purified aldose of purified aldose reductase, and analysis by g.l.c. revealed only the presence of unchanged monosaccharides. Addition of the aldose reductase inhibitor sorbinil to purified rat lens aldose reductase incubated with xylose in the presence of NADPH resulted in decreased xylitol production. However, aldose reductase inhibitors produced no effect in altering the rate of Nitro Blue Tetrazolium formation from either glucose or xylose, indicating that the observed inhibition in vitro does not result from a free radical-scavenger effect. PMID- 3030279 TI - Effects of insulin, glucagon, dexamethasone, cyclic GMP and spermine on the stability of phosphatidate phosphohydrolase activity in cultured rat hepatocytes. AB - Hepatocytes were preincubated with 10mM-glucagon and 100 microM-corticosterone to increase phosphatidate phosphohydrolase activity. Addition of 10 nM-glucagon or 100 microM-8-bromo cyclic GMP to a second incubation mixture that contained cycloheximide increased the half-life of the phosphohydrolase activity. Dexamethasone (100 nM) had no significant effect, but insulin (500 pM) or spermine (1 mM) decreased the half-life. None of these compounds altered the general rate of degradation of proteins labelled with [3H]leucine. There appears to be a specific control of the half-life of phosphatidate phosphohydrolase activity, which could contribute to its long-term regulation in the liver. PMID- 3030280 TI - Copper transport from Cu(I)-thionein into apo-caeruloplasmin mediated by activated leucocytes. AB - A study on the transfer of copper from Cu-thionein into apo-caeruloplasmin, using Cu-thionein that was previously oxidised by activated leucocytes, was performed. Cu(I)-thiolate oxidation was conveniently monitored by the progressive decline of the specific Cotton bands between 400 and 300 nm. The characteristic e.p.r. properties and NN-dimethyl-p-phenylenediamine oxidase activity indicated a successful formation of caeruloplasmin. Taking into account the simultaneous occurrence of leucocytes, apo-caeruloplasmin and Cu-thionein in blood plasma, such an interaction would favour a possible metabolic link between either copper protein. PMID- 3030277 TI - Characterization of specific V1a vasopressin-binding sites on a rat mammary tumour-cell line. AB - WRK 1, a cloned cell line derived from a rat mammary tumour, carries specific vasopressin-binding sites. Specific binding of 2-tyrosine-3H-labelled [8 lysine]vasopressin ([3H]vasopressin) was time-dependent, saturable and reversible. Scatchard-plot analysis of hormone binding indicated the presence of a single class of receptors with an equilibrium dissociation constant of 12.7 +/- 0.2 nM. The maximal binding capacity was 75 +/- 6 fmol/10(6) cells, which corresponds to approx. 45,000 sites per cell. Oxytocin and a highly potent oxytocin analogue were able to inhibit completely [3H]vasopressin binding, but, in this respect, they were far less potent than vasopressin. This clearly demonstrates the vasopressinergic nature of this receptor. Pharmacological studies using a series of 14 vasopressin or oxytocin analogues indicated that the ligand selectivity of the vasopressin receptor found on WRK 1 cells resembles that of the rat hepatocyte. This signifies that this vasopressin receptor is of the V1a subtype. This conclusion was confirmed by the observation that vasopressin did not influence the production of intracellular cyclic AMP in WRK 1 cells. PMID- 3030281 TI - Phosphorylation of purified bovine heart and rat liver 6-phosphofructo-2-kinase by protein kinase C and comparison of the fructose-2,6-bisphosphatase activity of the two enzymes. AB - Purified bovine heart 6-phosphofructo-2-kinase can be phosphorylated in the presence of protein kinase C and dephosphorylated by alkaline phosphatase; changes in phosphorylation state have no effect on enzyme activity. By contrast, the rat liver enzyme is a poor substrate for protein kinase C. Unlike the liver enzyme, which is bifunctional and is phosphorylated by fructose 2,6-[2 32P]bisphosphate, the heart enzyme contains 10 times less fructose 2,6 bisphosphatase activity and is phosphorylated at a slower rate and to a lesser extent than the liver enzyme. Both rat liver and bovine heart enzymes catalyse a similar exchange reaction between [U-14C]ADP and ATP. PMID- 3030282 TI - Specificity of protein phosphatases in the dephosphorylation of protein kinase C. AB - Protein kinase C can autophosphorylate in vitro and has also been shown to be phosphorylated in vivo. In order to investigate the factors that may determine the phosphorylation state of protein kinase C in vivo, we determined the ability of the ATP + Mg2+-dependent phosphatase and the polycation-stimulated (PCS) phosphatases to dephosphorylate protein kinase C in vitro. These studies show that all the oligomeric forms of the PCS phosphatases (PCSH1, PCSH2, PCSM and PCSL phosphatases) are effective in the dephosphorylation of protein kinase C, showing 34-82% of the activity displayed with phosphorylase a as substrate. In contrast both the catalytic subunit of the PCS phosphatase and that of the ATP+Mg2+-dependent phosphatase showed only weak activity with protein kinase C as substrate. All these phosphatases, however, were activated by protamine (Ka 14-16 micrograms/ml) through what appears to be a substrate-directed effect. The relative role of these phosphatases in the control of protein kinase C is discussed. PMID- 3030283 TI - A cyclic nucleotide-independent protein kinase in Leishmania donovani. AB - Leishmania donovani promastigotes labelled for 2 h with 32Pi incorporated radioactivity into at least 21 different proteins, as determined by SDS/polyacrylamide-gel electrophoresis. Pulse-chase studies with 32Pi demonstrated that the labelled proteins were in a dynamic state: some radiolabelled proteins rapidly disappeared and others appeared after the chase. The possibility of an ectokinase on the parasite was examined; incubation of intact parasites for 10 min at 25 degrees C in an osmotically buffered medium containing [gamma-32P]ATP, but not [alpha-32P]ATP, resulted in the labelling of 10 different protozoal proteins, presumably localized to the surface of the organism's plasma membrane. Intact promastigotes also catalysed the transfer of 32P from [gamma-32P]ATP to histones. The histone-dependent kinase was solubilized by repeated freezing and thawing, and sonication, and purified 118-fold by chromatographing the high-speed (200,000 g, 1 h) supernatant fraction on QAE Sephadex, Sephadex G-150 and hydroxyapatite columns. The kinase eluted as a single activity peak from all three columns. The partially purified histone dependent kinase had the following properties: pH optimum, 7.0; optimum temperature, 37 degrees C; Km for mixed calf thymus histone, 0.15 mM; Km for ATP, 0.8 mM; preferred fractionated histone acceptors, H2b greater than H4 greater than H2a greater than H3 (H1 does not serve as an acceptor); optimum activity required 10-20 mM-Mg2+; inhibited 50-80% by 0.01 mM- and 1 mM-Ca2+; activity was not stimulated by calmodulin, cyclic AMP (1 mM) or cyclic GMP (1 mM) nor inhibited by a cyclic AMP-dependent protein kinase inhibitor (50 micrograms/assay); apparent Mr 75,000, as determined by Sephadex G-150 gel filtration chromatography; phosphorylated exclusively serine residues. Protein kinase activity was low in the early exponential phase of the growth curve and increased 6-fold upon entry into the stationary phase. PMID- 3030284 TI - Complete quantification of the total concentration of rat skeletal-muscle Na+ + K+-dependent ATPase by measurements of [3H]ouabain binding. AB - In the standard [3H]ouabain-binding assay for quantification of the Na,K-ATPase (Na+ + K+-dependent ATPase) concentration in rat skeletal muscles, samples are incubated for 2 X 60 min in 1 microM-[3H]ouabain at 37 degrees C followed by a wash-out for 4 X 30 min at 0 degree C. To obtain accurate determinations, values determined by this standard assay should be corrected for non-specific uptake and retention of [3H]ouabain (11% overestimation), loss of specifically bound [3H]ouabain during wash-out (21% underestimation), evaporation from muscle samples during weighing (4% overestimation), impurity of [3H]ouabain (5% underestimation) and incomplete saturation of [3H]ouabain binding sites (6% underestimation). Thus corrected the standard [3H]ouabain-binding assay determines the total Na,K-ATPase concentration. Hence, in the soleus muscle of 12 week-old rats the total [3H]ouabain-binding-site concentration is 278 +/- 20 pmol/g wet wt. This is at variance with the evaluation of the Na,K-ATPase concentration from Na,K-ATPase activity measurements in muscle membrane fractions, where the recovery of Na,K-ATPase is only 2-18%. Quantification of the total Na,K-ATPase concentration is of particular importance since it is a prerequisite for the discussion of quantitative aspects of the Na,K-ATPase. PMID- 3030285 TI - Inhibition of gluconeogenesis by hypoglycin in the rat. Evidence for inhibition of glucose-6-phosphatase in vivo. AB - Treatment of rats with hypoglycaemic doses of hypoglycin has been shown to abolish the relative detritiation of [2-3H,U-14C]glucose [Osmundsen, Billington, Taylor & Sherratt (1978) Biochem. J. 170, 337-342], indicating that both the Cori and the glucose/glucose 6-phosphate cycles were inhibited in vivo. This inhibition was confirmed and, in addition, it was shown that the conversion in vivo of both [14C]lactate and [14C]fructose into glucose was decreased after hypoglycin treatment. These results suggest that hypoglycin poisoning results in the inhibition in vivo of glucose-6-phosphatase activity, which participates in the overall inhibition of gluconeogenesis and hypoglycaemia. Clofibrate feeding apparently protected the rats against the inhibition of the fructose-to-glucose conversion by hypoglycin. However, in isolated hepatocytes prepared from hypoglycin-treated rats, the conversion of [14C]fructose into glucose and the recycling of [2-3H,U-14C]glucose were not different from that in control hepatocytes. This suggests that the inhibition was lost during preparation of the hepatocytes. The direct measurement of glucose-6-phosphatase activity showed that it was inhibited when measured in concentrated, but not dilute, homogenates prepared from hypoglycin-treated rats. PMID- 3030287 TI - Studies on the photolytic breakdown of hydroperoxides and peroxidized fatty acids by using electron spin resonance spectroscopy. Spin trapping of alkoxyl and peroxyl radicals in organic solvents. AB - Spin trapping using 5,5-dimethyl-1-pyrroline N-oxide (DMPO) has been used to detect and distinguish between the carbon-centred, alkoxyl, and peroxyl radicals produced during the photolytic decomposition of hydroperoxides. Photolysis of tert-butyl and cumene hydroperoxides, and peroxidized fatty acids, in toluene, with low levels of u.v. light, is shown to lead to the initial production of alkoxyl radicals by homolysis of the oxygen-oxygen bond. Subsequent reaction of these radicals with excess hydroperoxide leads, by hydrogen abstraction, to the production of peroxyl radicals that can be detected as their corresponding adducts with the spin trap. Subsequent breakdown of these adducts produces alkoxyl radicals and a further species that is believed to be the oxidized spin trap radical 5,5-dimethyl-1-pyrrolidone-2-oxyl. No evidence was obtained at low hydroperoxide concentrations, with either the cumene or lipid alkoxyl radicals, for the occurrence of beta-scission reactions; the production of low levels of carbon-centred radicals is believed to be due to the alternative reactions of hydrogen abstraction, ring closure, and/or 1,2 hydrogen shifts. Analogous experiments with 3,3,5,5-tetramethyl-1-pyrroline N-oxide (TMPO) led only to the trapping of alkoxyl radicals with no evidence for peroxyl radical adducts, this is presumably due to a decreased rate of radical addition because of increased steric hindrance. PMID- 3030286 TI - Cloning and sequencing of full-length cDNA encoding the precursor of human complement component C1r. AB - The sequencing of human liver cDNA clones encoding the entire C1r precursor protein has confirmed the previously determined peptide sequence and has shown that there is a leader peptide which is 17 amino acids long. A residue tentatively identified as beta-hydroxyaspartic acid [Arlaud, Willis & Gagnon (1986) Biochem. J., in the press] located in the C1r A-chain, within an epidermal growth-factor consensus sequence, was found to be encoded as asparagine. Two sequence elements, tandemly located in the A-chain, are related to a sequence widespread among proteins which interact with C3b or C4b. Structural comparisons between different clones indicate that multiple polyadenylation sites are responsible for the length heterogeneity observed for C1r mRNA from liver and Hep G2 cells. PMID- 3030288 TI - The control of CTP:choline-phosphate cytidylyltransferase activity in pea (Pisum sativum L.). AB - Several possible control mechanisms for CTP:choline-phosphate cytidylyltransferase (EC 2.7.7.15) activity in pea (Pisum sativum L.) stems were investigated. Indol-3-ylacetic acid (IAA) treatment of the pea stems decreased total cytidylyltransferase activity but did not affect its subcellular distribution. Oleate (2 mM) caused some stimulation of enzyme activity by release of activity from the microsomal fraction into the cytosol, but neither phosphatidylglycerol nor monoacyl phosphatidylethanolamine had an effect on activity or subcellular distribution. A decrease in soluble cytidylyltransferase protein concentrations was found in IAA-treated pea stems, but this was not sufficient to account for all of the decrease in cytidylyltransferase activity. A 50% inhibition of enzyme activity could be obtained with 0.2 mM-CMP, which indicated possible allosteric regulation. Similar inhibition was obtained with 1.5 mM-ATP, but other nucleotides had no effect. The cytidylyltransferase enzyme protein was not directly phosphorylated, and the inhibition with 1.5 mM-ATP occurred with the purified enzyme, thus excluding an obligatory mediation via a modulator protein. The results indicate that the cytosolic form of cytidylyltransferase is the most important in pea stem tissue and that the decrease in cytidylyltransferase activity in IAA-treated material appears to be brought about by several methods. PMID- 3030289 TI - Thiyl free radicals and the oxidation of ferrocytochrome c. Direct observation of coupled hydrogen-atom- and electron-transfer reactions. AB - Absolute rate constants for the reaction of ferrocytochrome c with the thiyl radicals derived from cysteine, GSH, penicillamine and N-(2 mercaptopropionyl)glycine were measured by using the technique of pulse radiolysis. The reaction is believed to occur through a one-electron-transfer process, in agreement with the hypothesis that thiols may act as catalysts linking hydrogen-atom- and electron-transfer reactions. PMID- 3030290 TI - Comparison of human stromelysin and collagenase by cloning and sequence analysis. AB - A comparison of the cDNA-derived amino acid sequences of human stromelysin and collagenase with the N-terminal sequences of purified enzymes reveals that these metalloproteinases are highly conserved and that they are secreted as proenzymes. A putative zinc-binding site was identified by its homology with the zinc chelating sequence of thermolysin. These sequences permitted the identification of: transin, a protein induced in rat fibroblasts either exposed to growth factors or transformed by oncogenic viruses, as the rat homologue of stromelysin, and XHF1, a protein induced in human fibroblasts after treatment with tumourigenic agents, as collagenase. PMID- 3030291 TI - A receptor binding assay for endoxin in human urine. AB - Endoxin is an endogenous substance known to participate in the regulation of the sodium balance and hypertension. Its chemical nature remains elusive. Based on its capacity to specifically bind to Na-K ATPase receptors we describe a receptor assay for its measurement in human urine. The endoxin was extracted by methanol and desalted on a silicagel column with a mixture of chloroform-ethanol. Calibration curves have been established by the transformation of the weight of crude extract in actual content of active material expressed in nmol/l as calculated from Scatchard plots analysis. Normal values are given for subjects on regular salt diet. Changes in endoxin urinary elimination after an oral salt load are outlined. PMID- 3030292 TI - Genomic clone of hst with transforming activity from a patient with acute leukemia. AB - We have previously reported the identification of a novel transforming gene, hst, in DNA samples taken from human stomach cancers and a noncancerous portion of stomach. Five clones, containing the genomic hst gene, were isolated from a human cosmid library constructed from leukocyte DNA from a patient with acute leukemia. All clones possessed transforming activity when transfected to NIH3T3 cells. From one clone, an 8.7 kb BamHI fragment was subcloned into pBR322, and this subclone was active in transforming NIH3T3 cells. This is the first isolation of transformation-competent genomic hst clones directly from a human genomic library, that is, without prior passage through NIH3T3 cells. PMID- 3030293 TI - Identification of the Rts 1 DNA fragment responsible for temperature sensitive growth of host cells harboring a drug resistant factor Rts 1. AB - We have placed a kanamycin resistance SalI fragment (3.65 Kb) from the drug resistance factor Rts1 into pUC19 and pBR322. These chimeric plasmids containing the kanamycin resistance fragment from Rts1 cause temperature sensitive growth in E coli. The orientation of the kanamycin resistance fragment in the vector plasmids does not influence this phenotype. PMID- 3030294 TI - Base liberation from nucleotides by superoxide and intramolecular enhancement effect of phosphate group. AB - The reaction of nucleotides with superoxide gave the corresponding nucleobases in good yield. The base-release reaction of nucleotides due to superoxide was examined and compared to that of nucleosides. Hence, the phosphate moiety greatly enhances the yields, in particular those of adenine from adenosine monophosphates. Superoxide in combination with the phosphate moiety has been revealed to be more active than superoxide alone. The phosphate peroxy radical, generated in situ from superoxide and phosphate, seems to be the species responsible for the formation of the free bases. PMID- 3030295 TI - Effects of platelet activating factor on the chemotaxis of normodense eosinophils from normal subjects. AB - Highly purified eosinophils were obtained from normal subjects, and their chemotactic responses to platelet activating factor (PAF) were evaluated. PAF induced both eosinophil chemotactic and chemokinetic responses, and was 100 fold more potent eosinophil chemotactic factor as compared to eosinophil chemotactic factor of anaphylaxis. On the other hand, leukotriene B4 did not show any eosinophil chemotactic activity. A selective PAF antagonist, BN52021, inhibited eosinophil chemotaxis in a dose dependent manner. Preincubation of eosinophils with PAF induced the deactivation of eosinophils for further chemotactic responses to PAF. These findings suggest that PAF is a potent chemotactic factor for normal eosinophils. PMID- 3030296 TI - Enzymatic conversion of leukotriene B4 to 6-trans-leukotriene B4 by rat kidney homogenates. AB - A novel isomerase reaction leading to conversion of leukotriene B4 to its 6-trans isomer was detected in rat kidney homogenates. The structure of the metabolite was determined by high performance liquid chromatography, ultraviolet spectrometry and gas-liquid chromatography-mass spectrometry. A recent report has shown that 6-trans-leukotriene B4 is transformed to a dihydro metabolite (6,7- or 10,11-dihydro 6-trans-leukotriene B4) and further omega-hydroxylated [Powell, W.S. (1986) Biochem, Biophys. Res. Commun. 136, 707-712]. The leukotriene B4 6 isomerase reaction reported here may therefore provide the first step in a novel pathway of biological degradation of leukotriene B4. PMID- 3030297 TI - Epidermal growth factor (EGF) stimulates inositol trisphosphate formation in cells which overexpress the EGF receptor. AB - EGF is a low molecular weight polypeptide hormone which acts as a regulator of cell growth and differentiation. The A-431 cell line has been used frequently to examine receptor-mediated biochemical effects of EGF, since this cell line has an increased (20-50 fold) level of EGF receptors. We have utilized A-431 cells to examine the influence of EGF on formation of an intracellular second messenger, inositol, 1,4,5-trisphosphate (Ins-1,4,5-P3), and other inositol phosphates. The results show that EGF induces rapid formation of Ins-1,4,5-P3 as well as Ins 1,3,4-P3 and Ins-1,3,4,5-P4. There is a concurrent decrease in the level of the lipid precursor for Ins-1,4,5-P3, phosphatidylinositol 4,5-biphosphate (PIP2). Furthermore, we have examined five other cell lines that overexpress the EGF receptor and find that EGF treatment induces formation of inositol polyphosphates in those cell lines also. PMID- 3030298 TI - Role of carbohydrates in the viscosity and permeability of gastric mucin to hydrogen ion. AB - The effect of carbohydrate removal on the viscosity of gastric mucin and its ability to impede the diffusion of hydrogen ion was investigated. The mucin, purified from dog gastric mucus, was subjected to partial or extensive deglycosylation with specific exoglycosidases and then used in the measurements. The obtained results revealed that removal of peripheral fucose or N acetylglucosamine caused in each case only about 5% reduction of the glyco protein viscosity. An 18% drop in the viscosity, however, occurred following removal of sialic acid, while extensive deglycosylation (removal of 86% carbohydrate) reduced the glycoprotein viscosity by 40%. The ability of mucin to retard the diffusion of hydrogen ion increased by 7% following removal of fucose or N-acetylgalactosamine, a 28% increase was obtained following removal of sialic acid, while the permeability to hydrogen ion of the extensively deglycosylated glycoprotein decreased by 42%. The results suggest that carbohydrates contribute significantly to the viscoelastic and permselective properties of gastric mucin. PMID- 3030299 TI - Tumor necrosis factor stimulates gelatinase and collagenase production by granulation tissue in culture. AB - Tumor necrosis factor (TNF, 2.6 X 10(-11)-10(-9) M) caused an increase in the production of active gelatinase and latent collagenase by granulation tissue in culture. As determined by SDS-substrate polyacrylamide gel electrophoresis, granulation tissue produced mainly two species of gelatinase with molecular weights of 64 kDa and 57 kDa, and an additional 80-kDa gelatinase was produced by TNF treatment. The results suggest that TNF may play a role in a rapid collagen turnover in the carrageenin-induced granulation tissue in rats. PMID- 3030300 TI - Prevalence of polymorphic 21-hydroxylase gene (CA21HB) mutations in salt-losing congenital adrenal hyperplasia. AB - Using genomic restriction analysis of 14 unrelated patients with salt-losing congenital adrenal hyperplasia, we identified three different CA21HB mutation patterns: no detectable restriction fragment abnormalities (16/28 haplotypes), deletion of the active CA21HB gene (9/28), and apparent conversion of the active CA21HB gene to the pseudogene CA21HA (3/28). CA21HB gene deletion was associated with HLA-Bw47 in 6 haplotypes and with absent C4B expression in 7. A variety of HLA and C4 types was associated with the other mutations. Apparent conversion of CA21HB to CA21HA was identified by the disparity between the intensity ratios for the major TaqI and BglII hybridization fragments. PMID- 3030301 TI - Establishment of hybridoma secreting human monoclonal antibody against hepatitis B virus surface antigen. AB - The HAT (hypoxanthine, aminopterin, thymidine) sensitive and ouabain resistant human B lymphoblastoid cell line TAW-925 was obtained from 6-thioguanine resistant B lymphoblastoid cell line WI-L2. Hybridomas were obtained at a high frequency (10(-4)-10(-5) when TAW-925 was hybridized with cells transformed with Epstein-Barr virus. Using TAW-925 as a parental cell line, we have obtained a hybridoma which stably secretes human monoclonal antibody against hepatitis B virus surface antigen. PMID- 3030302 TI - Cardiac glycosides stimulate phospholipase C activity in rat pinealocytes. AB - Ouabain and related cardiac glycosides stimulate phospholipase C activity 5-fold in rat pinealocytes. The combined treatment of ouabain and norepinephrine, which also stimulates phospholipase C, produces an additive effect. The effects of either ouabain or norepinephrine are blocked by EGTA. However, there are notable differences. The stimulatory effect of ouabain is lost when extracellular Na+ is reduced to 20 mM and is not blocked by prazosin. In contrast, the stimulatory effect of norepinephrine is not blocked when extracellular Na+ is reduced to 20 mM but is blocked by prazosin. Ouabain appears to increase phospholipase C activity through a mechanism involving inhibition of Na+,K+-ATPase, and an accumulation of intracellular Na+ and Ca2+, not involving alpha 1-adrenoceptors. These findings raise the possibility that activation of phospholipase C might be a more general effect of cardiac glycosides. PMID- 3030303 TI - lon-sulA regulatory function affects the efficiency of transposition of Tn5 from lambda b221 cI857 Pam Oam to the chromosome. AB - In transposition of Tn5 from lambda b221 cI857 Pam Oam rex::Tn5 to the chromosome, lon mutants of Escherichia coli K12 are less efficient than lon+ parents. Reduced frequency of transposition of Tn5 is recovered to the wild type level by adding DL-pantoyl lactone, an agent which suppresses Lon- phenotypes. A suppressor mutation, sulA (or sfiA), suppresses the deficiency of transposition as well as other known lon-associated phenotypes, whereas a lon sulB (or sfiB) double mutant is phenotypically Lon+ but still transposition-deficient. An additive effect of the sulA genes on teh chromosome and on a high copy number plasmid is found in inhibiting transposition of Tn5. A hypothesis that the SulA product someway regulates transposition of Tn5 negatively is proposed. PMID- 3030304 TI - Isoenzyme-specific phosphorylation of cytochromes P-450 and other drug metabolizing enzymes. AB - A series of fourteen cytochrome P-450 isoenzymes was treated with three different protein kinases and found to divide into isoenzymes phosphorylated by both the cyclic AMP-dependent kinase and the calcium-phospholipid-dependent kinase (P-450 PB 3a and PB 2e), by none of these kinases (P-450 PB 1b, MC 1b, UT 1, and thromboxane synthase), and by either the cyclic AMP-dependent kinase (P-450 LM 2, PB 2d, and PB 3b) or the calcium-phospholipid-dependent kinase (P-450 PB 1a, PB 2a, MC 1a, LM 3c, and LM 4). Other components of the monooxygenase system, cytochrome P-450 reductase, cytochrome b5, cytochrome b5 reductase as well as microsomal epoxide hydrolase, were poor substrates for the kinases employed. On the other hand, glutathione transferases 1-2 and 4-4, but not 3-3, were relatively good substrates for the calcium-phospholipid-dependent kinase. PMID- 3030305 TI - Heterogeneity of beta-adrenoreceptors in guinea pig alveolar type II cells. AB - [3H]Dihydroalprenolol ([3H]DHA) binding to guinea pig alveolar type II cell membrane revealed the presence of both high (KD = 0.38 nM) and low (KD = 4.2 nM) affinity beta-adrenoreceptors. The low affinity site had a higher binding capacity (Bmax = 245.6 fmol/mg protein) than the high affinity site (Bmax = 71.7 fmol/mg protein). Displacement of [3H]DHA by practolol, a selective beta 1 agent, confirmed the existence of two species of adrenoreceptors, corresponding to 21% high affinity (beta 1) and 79% low affinity (beta 2), respectively. PMID- 3030307 TI - Occurrence, identification and possible significance of ornithine lipid in Thiobacillus ferrooxidans. AB - An ornithine containing aminolipid has been found in Thiobacillus ferrooxidans grown in ferrous sulfate medium, which was purified and estimated at four main phases of growth. GLC analysis of ornithine lipid has revealed the existence of mainly C18:1 and C22:1 fatty acids. The infrared spectra showed the existence of both amide and ester linkages in the aminolipid. The major ester linked fatty acid was C22:1. The interaction of ornithine lipid with membrane was investigated by delipidation of the membrane particles, which resulted in the perturbation of the activities of the three enzymes of iron oxidation system. The activities could be restored to the lipid depleted particles by preincubation with a dispersion of purified ornithine lipid together with coenzyme Q8. The kinetic parameters of the enzyme activities were also affected by delipidation which was significantly altered in the reconstituted particles by this lipid, thus indicating a possible role of ornithine lipid in iron oxidation system. PMID- 3030306 TI - Phosphorylation of brain muscarinic receptor: evidence of receptor regulation. AB - Muscarinic receptor, from porcine synaptic membrane, was purified by affinity chromatography. Molecular weight analysis by SDS-gel electrophoresis revealed one major peptide with an apparent Mr of 68 +/- 2 Kda. The purified receptor was phosphorylated by the catalytic subunit of cAMP-dependent protein kinase resulting in a concomitant loss in specific binding, and this loss was reversed by calcineurin. PMID- 3030308 TI - The expression of oncogenes in human developing liver and hepatomas. AB - Oncogene expression was examined in the human fetal liver and human hepatomas. Erb (B), erb (A+B), Ha-ras, myc, fos and fms oncogene expression elevated in certain stages of fetal liver development and in hepatoma as compared to the normal adult human liver. In contrast, rel, src, mos, sis, myb, Ki-ras and bas oncogenes showed no apparent change of their mRNA levels during fetal liver development and in hepatoma. Further study of erb B oncogene expression in human cirrhotic liver and hepatoma demonstrated a strong correlation between erb B expression and alteration of its gene structure. PMID- 3030309 TI - Dopamine binding to the alpha receptor in pregnant rabbit myometrium. AB - This study evaluated in vitro binding of dopamine ligands to myometrial alpha adrenoceptors. With cell membranes from pregnant rabbits, receptor radioligand binding studies utilizing [3H] dihydroergocryptine +/- dopamine demonstrated receptor affinity (KD) = 0.75 +/- 0.10 nM (+/- SEM) and density (Bmax) = 533.2 +/ 45.2 fM/mg protein. Similar studies utilizing phentolamine or apomorphine gave essentially identical results. Competition binding studies demonstrated steriospecific butaclamol binding, along with significant binding of haloperidol, spiperone, apomorphine, and bromoergocryptine. These observations provide a mechanism for the observed uterotonic effects of dopamine. PMID- 3030310 TI - Isolation of a human liver ligandin cDNA clone and demonstration of sequence homology at ligandin loci in rats and humans. AB - Using a monospecific antibody to the major cytosolic glutathione-S-transferase of human liver, we have isolated a cDNA clone from a human liver cDNA expression vector library in lambda gt11. The clone cross-hybridizes with a rat liver ligandin (glutathione-S-transferase 1-2) cDNA probe. The clone has an insert of 1.25 kb, a size sufficient to code for the 23 kilodalton subunit of human GST. Digestion of the insert with Hinf I produced three fragments (0.8 kb, 0.4 kb and 0.1 kb). A similar pattern of multiple bands was observed when rat liver GST1-2 cDNA probe was used for Southern blot analysis of Pst digests of rat and human genomic DNAs. These data suggest that these two functionally similar proteins exhibit sequence homology between their respective cDNAs and at ligandin loci, in spite of the lack of immuno-crossreactivity between them. PMID- 3030311 TI - Effect of platelet-activating factor on beta-adrenoceptors in human lung. AB - We examined the effect of platelet-activating factor (PAF) on beta-adrenoceptors in the membranes of human lung and the functional responses to methacholine, histamine and isoproterenol in human lung parenchyma and tracheal strips. Preincubation of the human lung slices with 0.1 microM PAF decreased the density of beta-adrenoceptors without a change in the affinity. In the functional studies, a subthreshold dose of PAF (0.1 microM) significantly reduced the potency of isoproterenol to reverse methacholine or histamine-induced contraction. BN 52021, a PAF antagonist, protected against these PAF-induced changes. These findings suggest that PAF-induced down-regulation of beta adrenoceptors could be a mechanism of the non-specific airway hyperreactivity in asthma. PMID- 3030312 TI - Suppression of matrix protein synthesis by herpes simplex virus in bovine smooth muscle cells. AB - We investigated the effect of herpes simplex virus type 1 infection on the synthesis of extracellular matrix proteins by bovine smooth muscle cells. The cells supported viral replication, which reached maximum at 24 hrs post infection. Uninfected and infected (multiplicity of infection of 5 or 20) cultures of smooth muscle cells were labeled with [14C]proline at five hrs post infection. Incorporation of radioactivity into non-dializable protein was determined following electrophoresis of the cell-matrix and medium fractions. The synthesis of fibronectin and collagen types I and III was almost completely suppressed. The data suggest that HSV-1 infection of smooth muscle cells in vitro alters extracellular matrix synthesis. PMID- 3030313 TI - Plasma immunoreactive atrial natriuretic factor (IR-ANF) increases markedly after alpha 2-adrenergic stimulation with clonidine in normally-hydrated rats. AB - Clonidine, an alpha 2-adrenergic agonist, induced a marked, dose-related increase of plasma IR-ANF in normally-hydrated rats. Maximal ANF release was observed at 10 min after injection of 50 micrograms clonidine, rising from 40.5 +/- 4.6 pg/ml (X +/- SEM) to 1064.4 +/- 22.4 pg/ml. This effect on plasma IR-ANF was partially blocked by pretreatment with 0.8 mg naloxone, whereas synthetic Arg8-vasopressin (AVP) did not inhibit clonidine's action. These findings indicate that increased ANF release may be involved in the mechanism of clonidine-induced diuresis. The clonidine's effect on ANF release may be mediated via activation of opioid receptors besides stimulation of alpha 2-adrenergic receptors. PMID- 3030314 TI - Do myc, fos and E1A function as protein phosphatase inhibitors? AB - The oncogenic proteins myc, fos and E1A bear striking resemblance to protein phosphatase inhibitors 1 and 2. Both sets of proteins possess several regions rich in proline (P), glutamic acid (E), serine (S) and threonine (T). In addition to PEST sequences four of the five proteins contain clusters of arginine-arginine pairs. On the basis of these similarities, I suggest that myc, fos and E1A are protein phosphatase inhibitors. PMID- 3030315 TI - Binding of zinc(II) to beta-D-fructose 2,6-bisphosphate. AB - The formation of a complex between Zn(II) and beta-D-fructose 2,6-bisphosphate was shown because the latter compound: activated bis(5'-guanosyl)tetraphosphatase (EC 3.6.1.17) and dinucleoside triphosphatase (EC 3.6.1.29) only to the extent that they could be inhibited by Zn(II); increased the consumption of Zn(II) necessary to titrate to an end point a solution of the metallochromic indicator eriochrome black T; coeluted with Zn(II) in a gel filtration column capable of resolving them if unbound. Neither of those effects was shown by D-fructose 1,6 bisphosphate under the same conditions. PMID- 3030316 TI - Anomeric specificity of glucose effect on cAMP, fructose 1,6-bisphosphatase, and trehalase in yeast. AB - The addition of beta-D-glucose (final concentration, 50 mM) to a cell suspension of Saccharomyces cerevisiae in stationary phase caused a rapid 4-fold increase in the concentration of cAMP, while a 2-fold increase of cAMP was observed by the addition of alpha-D-glucose. beta -D-Glucose was also more effective than alpha-D glucose in the inactivation of fructose 1,6-bisphosphatase and the activation of trehalase. These results, taken together with the previous report that alpha-D glucose is transported more rapidly than beta-D-glucose in Saccharomyces cerevisiae, do not support the view currently proposed by some investigators that cotransport of D-glucose with protons causes the depolarization of the cell membrane, resulting in the activation of adenylate cyclase. The present data, however, provides supporting evidence for the view that cAMP-dependent protein kinase is implicated in the inactivation of fructose 1,6-bisphosphatase and the activation of trehalase. PMID- 3030317 TI - A strong homology exists between the active T-cell binding gp120 octapeptide of human immunodeficiency virus and the subtilisin cleavage peptide of bovine ribonuclease A. AB - A homology has been found between an octapeptide involved in attachment of the human immunodeficiency virus to helper/inducer T cells and an octapeptide segment of bovine pancreatic ribonuclease A. This segment (residues 19-26) contains the sites for subtilisin cleavage of this enzyme into the S-peptide and S-protein. From the X-ray crystal structure of ribonuclease, this sequence is known to be exposed to solvent and interacts little with the rest of the protein. A structure for the human immunodeficiency virus attachment peptide can be deduced from this homology, as a well-defined structure has been determined for this sequence in ribonuclease. This can be readily accomplished using previously developed computer methods based upon conformational energy calculations. The calculated structure for human immunodeficiency virus peptide is identical to the ribonuclease segment (19-26) in backbone conformation. It is stabilized by internal interactions of nonpolar residues, and by exposure of polar hydroxyl groups. The results suggest that the T-cell human immunodeficiency virus receptor may be hydrophilic in nature and that conservation of the sequence in two presumably functionally unrelated proteins is related to the need for conservation of exposed structure. PMID- 3030318 TI - Expression of human atrial natriuretic polypeptide gene in Cos 7 cells. AB - Cos 7 cells transfected with human atrial natriuretic polypeptide (hANP) gene with SV40 enhancer and replication origin sequences expressed hANP gene. The expressed RNA was indistinguishable from native hANP mRNA and the transcribed protein seemed to be properly processed to alpha-hANP and beta-hANP. This system provides a useful approach to investigate the processing of hANPs and the structure-function relationship of amino acid sequences of hANPs. PMID- 3030319 TI - Repetitive region of calpastatin is a functional unit of the proteinase inhibitor. AB - A cDNA portion coding for one of the repetitive regions of pig heart calpastatin (107 kDa) was subcloned into E. coli plasmid pUC119 to express the portion of the proteinase inhibitor gene in bacteria. The expressed protein was a chimaeric protein whose calpastatin segment (130 amino acid residues) was fused with an amino-terminus portion (7 amino acid residues) of beta-galactosidase. The chimaeric protein could inhibit proteolytic activity of calpain (Ca2+-dependent cysteine proteinase), and maintained properties of the authentic calpastatin concerning inhibition specificity and heat stability. These findings led us to conclude that the repetitive region is a functional unit of the proteinase inhibitor. PMID- 3030320 TI - Specific binding of leukotriene B4 to guinea pig lung membranes. AB - We have demonstrated binding sites for LTB4 in guinea pig lung membranes. Binding of [3H]-LTB4 was of high affinity (Kd = 0.76 nM), saturable and linear with protein concentration (0.2-1.2 mg/ml). Scatchard and Hill's plot analysis indicated a single class of binding site with a Hill's coefficient of 0.99 +/- 0.08 (n = 4). [3H]-LTB4 was unmetabolized during incubation with membrane preparations, as indicated by high performance liquid chromatography. Divalent cations such as Mg2+ and Ca2+ enhanced binding capacity without changing the Kd. Na+ ions decreased binding in a concentration-dependent manner. Guanine nucleotides, GTP, GTP gamma S and Gpp(NH)p also decreased the number of binding sites. Finally, competition experiments demonstrated the following order of potency for displacement of [3H]-LTB4 from its receptor site: LTB4 greater than 20-OH-LTB4 much greater than 20-COOH-LTB4 = 6-trans-12-epi-LTB4 greater than LTC4 = LTD4 = 5-HETE. These data indicate that a specific LTB4 receptor, in addition to the previously documented LTC4 and LTD4 receptors, exists in guinea pig lung. PMID- 3030321 TI - Activation of mineralocorticoid agonist and antagonist specific receptors from rat kidney. AB - The kinetics of saturation, as well as of denaturation, confirm the existence of two distinct mineralocorticoid receptor populations one each for the agonist aldosterone (MR2) and the antagonist RU 26752 (MR3) in rat kidney. Receptor activation in vitro was dependent upon the buffer, progressed just as well in the presence of the agonist and the antagonist, and was inhibited by molybdate. These necessitate a reassessment of both the importance of receptor activation in vitro and its possible contribution to hormone action in vivo. PMID- 3030322 TI - Drug metabolism by the human hepatoma cell, Hep G2. AB - The human liver-derived cell line, Hep G2, has aryl hydrocarbon hydroxylase and 7 ethoxycoumarin o-de-ethylase activities. Partial purification of cytochrome P-450 from Hep G2 cells provided spectral evidence of this hemeprotein in the purified fraction. These results suggest that Hep G2 cells will be useful for the study of cytochrome P-450 and the regulation of mixed function oxidase activities in liver cells of human origin. PMID- 3030323 TI - Biological activities of catfish glucagon and glucagon-like peptide. AB - The ability of catfish glucagon and glucagon-like peptide to bind and activate mammalian glucagon receptors was investigated. Neither catfish peptide binds to glucagon receptors of rat liver, hypothalamus or pituitary. Neither stimulates adenylate cyclase activity in liver membranes. Catfish glucagon fails to activate adenylate cyclase in hypothalamic or pituitary membranes in contrast to mammalian glucagon. However, catfish glucagon-like peptide does stimulate hypothalamic and pituitary adenylate cyclase (EC50 approximately 1 pM) possibly through mammalian glucagon-like peptide receptors. PMID- 3030324 TI - Phospholipid metabolism and stimulus-response coupling. PMID- 3030325 TI - The influence of the antiviral drugs amantadine and rimantadine on erythrocyte and platelet membranes and its comparison with that of tetracaine. AB - The influence of the antivirus drugs amantadine and rimantadine and of the anionic analogue 1-adamantane-carboxylic acid on a range of properties of human erythrocyte membrane and of thrombocytes has been compared with the effect of the local anaesthetic tetracaine. At low antiviral drug concentrations the abilities of the drugs to induce erythrocyte shape change and suppress osmotic haemolysis were quantitatively proportional to their clinical potency (rimantadine more effective than amantadine at the same concentration). Rimantadine was also more effective than amantadine in suppressing influenza virus-erythrocyte fusion and viral induced haemolysis. The antiviral drug effects were qualitatively similar to those induced by tetracaine. At the quantitative level, tetracaine was more efficient than the antiviral drugs in inhibiting osmotic haemolysis, virus membrane fusion and platelet aggregation. In the absence of any specificity of the antiviral drug effects we argue for a lysosomotropic mode of drug action, i.e. that the drugs modify virus-membrane interactions by changing the endosomal or lysosomal pH. PMID- 3030326 TI - The inhibitory effects of some iodonium compounds on the superoxide generating system of neutrophils and their failure to inhibit diaphorase activity. AB - I have recently reported the inhibition of the neutrophil superoxide generating oxidase by very low concentrations of diphenylene iodonium (A. R. Cross and O. T. G. Jones, Biochem. J. 237, 111, 1986). Here I report on the sensitivity of the oxidase to two other iodonium compounds, iodonium thiophen and iodonium biphenyl. In addition, the lack of inhibition of dye reductase activity in a solubilized preparation of the oxidase is described suggesting that the superoxide forming enzyme system of neutrophils does not possess an intrinsic dye reductase activity. PMID- 3030327 TI - Muscarinic receptor stimulated phosphoinositide turnover in cardiac atrial tissue. AB - The properties of muscarinic agonist stimulated phosphoinositide turnover in canine atrial slices were investigated. In slices prelabeled with 32Pi, carbachol stimulated a 20-30% decrease of 32P-labeled phosphatidylinositol 4'-phosphate (PIP) and phosphatidylinositol 4',5'-bisphosphate (PIP2) content within 10-15 sec. This was followed by a resynthesis of these lipids to control levels after 30 sec. Carbachol-stimulated PIP and PIP2 turnover was followed by a relatively slower increase in 32P incorporation into phosphatidylinositol (PI) and phosphatidic acid (PA) which was maximal after 5-10 min. Carbachol increased 32P labeling of PA and PI in most regions of right and left atria with equal effectiveness. Muscarinic receptor stimulated increases in PA and PI labeling showed high specificity for certain muscarinic agonists and, unlike most tissues, this muscarinic receptor mediated phospholipid effect was dependent on extracellular calcium. Carbachol did not increase 32P incorporation into PA and PI if Mn2+, Co2+, Mg2+, or La3+ was substituted for extracellular Ca2+. Unlike other muscarinic agonists, acetylcholine increased 32P incorporation into phosphatidylcholine in addition to PA and PI. Low concentrations of calcium channel blockers, verapamil, nifedipine or diltiazem, did not block carbachol stimulated changes in PA and PI labeling in the presence of Ca2+; however, higher concentrations (greater than or equal to 10 microM) of verapamil increased PA and PI labeling. Ouabain enhanced carbachol-stimulated 32P incorporation into PA but attenuated incorporation into PI. The mechanisms associated with the actions of these agents on phospholipid metabolism and their possible physiological significance are discussed. PMID- 3030328 TI - Actions of antisecretory agents on proton transport in hog gastric microsomes. AB - The properties of K+-stimulated ATP hydrolysis (K+-ATPase) and vesicular accumulation of H+ (H+ accumulation) in hog gastric microsomes were investigated. The microsomes consisted of smooth surfaced vesicular particles, 70-300 nm in diameter. Both the activities of ATPase and the vesicular accumulation of H+ were stimulated by K+ in the presence of Mg2+, and enhanced by the K+-ionophore, valinomycin. However, there were differences in regulation of K+-ATPase and H+ accumulation by K+ ions, i.e. K+ at concentrations higher than 10 mM decreased K+ ATPase activity but further enhanced H+ transport. This observation suggests that the two reactions are partly independent. The H+ accumulation was inhibited by omeprazole, fenoctimine, spermine, and NaSCN, but not by cimetidine, prostaglandin E2, and atropine. The inhibitory effect of omeprazole on H+ accumulation paralleled the inhibition of K+-ATPase, while fenoctimine, spermine, and NaSCN suppressed H+ accumulation, without inhibiting K+-ATPase, under appropriate concentrations. In addition, the spontaneous diffusion of H+ across the microsomal membrane was markedly enhanced by fenoctimine, but not by the other agents used. These results indicate that omeprazole inhibits H+ accumulation by inhibiting K+-ATPase, fenoctimine suppresses H+ accumulation mainly by increasing the loss of accumulated H+ from the microsomal vesicles, spermine and NaSCN reduce H+ accumulation by inhibiting the transport of H+ into microsomal vesicles. PMID- 3030330 TI - Cromolyn inhibition of protein kinase C activity. PMID- 3030329 TI - In vitro metabolism of etoposide (VP-16-213) by liver microsomes and irreversible binding of reactive intermediates to microsomal proteins. AB - We have studied the metabolism of VP-16-213 (etoposide, VP-16), an antitumor agent, by mouse liver microsomes to reactive intermediates and the subsequent covalent binding to microsomal proteins. This metabolism was shown to involve the O-demethylation of VP-16 and resulted in the formation of a 3',4'-dihydroxy derivative (DHVP-16) which was identified by both HPLC and mass spectrometry. The formation of DHVP-16 was cytochrome P-450-mediated as indicated by its dependence on NADPH, its increased production following treatment of mice with phenobarbital, and its marked inhibition by SKF-525A and piperonyl butoxide. Furthermore, DHVP-16 formation required oxygen. Microsomal incubation of VP-16 resulted in an irreversible binding of the drug to the proteins, which was also shown to be cytochrome P-450 dependent. The covalent binding of the VP-16 metabolite(s) was inhibited by DHVP-16 in a dose-dependent fashion, suggesting that the reactive intermediates that bound to proteins were derived from DHVP-16. Electron spin resonance studies indicated that the same semiquinone radical was formed during enzymatic (oxidation or reduction) metabolism of DHVP-16 and the o quinone derivative of VP-16 (VP-16-Q). VP-16-Q and its semiquinone radical are suggested to be the bioalkylating species. PMID- 3030331 TI - Role of hepatic microsomal and purified cytochrome P-450 in one-electron reduction of two quinone imines and concomitant reduction of molecular oxygen. AB - The possible role of cytochrome P-450 in one-electron reduction of quinoid compounds as well as in the formation of reduced oxygen species was investigated in hepatic microsomal and reconstituted systems of purified cytochrome P-450 and purified NADPH-cytochrome P-450 reductase using electron spin resonance (ESR) methods. Two compounds were selected as model compounds: N-acetyl parabenzoquinone imine (NAPQI) and 3,5-dimethyl-N-acetyl-para-benzoquinone imine (3,5-dimethyl-NAPQI). Both compounds could be reduced by oxyhaemoglobin, the semiquinones formed were detectable by ESR and did not reduce molecular oxygen. Both NAPQI and 3,5-dimethyl-NAPQI underwent one-electron reduction in microsomal systems and in fully reconstituted systems of cytochrome P-450 and NADPH cytochrome P-450 reductase under anaerobic and aerobic conditions. In both incubation systems the semiquinone formation was diminished under aerobic circumstances and concomitant reduction of oxygen occurred, leading to the formation of hydrogen peroxide and hydroxyl free radicals. Both the reduction of the quinone imines and the reduction of oxygen were found to be cytochrome P-450 dependent. Both activities of cytochrome P-450 may also be involved in the bioactivation of other compounds with quinoid structural elements, like many chemotherapeutic agents. PMID- 3030332 TI - The influence of enalapril or spironolactone on experimental cyclosporin nephrotoxicity. AB - Adult Sprague-Dawley rats treated daily for 14 days with 50 mg/kg cyclosporin A (CsA) exhibited nephrotoxicity, characterized by reduced glomerular filtration rate, decreased urinary sodium and potassium flow, tubular enzymuria and proximal tubular structural damage. Elevations in plasma renin activity (PRA) were observed on day 4, but returned to normal within 7 days. Co-treatment of animals for the 14 day period with enalapril (8 mg/kg/day), a potent inhibitor of angiotensin converting enzyme (ACE), or spironolactone (25 mg/kg/day), the distal tubular antagonist of aldosterone, reduced the nephrotoxicity, although PRA remained elevated. Neither enalapril nor spironolactone affected circulating CsA levels. These data suggest that the action of aldosterone on the distal tubule may be important in the pathogenesis of CsA nephrotoxicity. PMID- 3030334 TI - Comparative study of [3H]glutamate binding sites in rat retina and cerebral cortex. PMID- 3030333 TI - Inhibition by reactive aldehydes of superoxide anion radical production from stimulated polymorphonuclear leukocytes and pulmonary alveolar macrophages. Effects on cellular sulfhydryl groups and NADPH oxidase activity. AB - Alpha,beta-unsaturated aldehydes such as acrolein (ACR) and crotonaldehyde (CRO) have been shown previously in our laboratory to inhibit the production of superoxide anion radical (O2-) by stimulated phagocytic cells in vitro in a dose related manner. Based on the known reactivity of these compounds towards cellular sulfhydryls (SH), the present studies were aimed at investigating cellular SH status in relation to O2- production. Plasma membrane surface SH groups were measured using carboxypyridinedisulfide and monitoring the resultant formation of mixed disulfides through assay of thione released into the supernatant fraction. Intracellular non-protein sulfhydryls were measured using 5,5'-dithiobis-2 nitrobenzoic acid. In both human polymorphonuclear leukocytes (PMN) and rat pulmonary alveolar macrophages (PAM) there was a dose-related decrease in surface SH and soluble SH after ACR and CRO treatment. Propionaldehyde, a three-carbon saturated aldehyde, was without effect. The decrease in surface SH was greater than the decrease in soluble SH. In addition, in PMN and PAM preincubated with 5 40 microM ACR, there was a dose-related inhibition in the rate of O2- production with no effect on the lag time as measured by cytochrome c reduction. In stimulated PMN, there was a dose-related decrease in the rate after addition of 5 40 microM ACR. These data suggest that changes in SH status by reactive aldehydes can modulate the activity of the plasma membrane NADPH oxidase responsible for O2 production. PMID- 3030335 TI - Assessment of kininases in rheumatic diseases and the effect of therapeutic agents. AB - Bradykinin is degraded in human plasma by a carboxypeptidase to yield desArg9 bradykinin (DBK) which is then digested by angiotensin-converting enzyme (ACE) to the pentapeptide Arg-Pro-Pro-Gly-Phe and the tripeptide Ser-Pro-Phe. We have studied the rate of kinin degradation by each of these enzymes in patients with rheumatoid arthritis (RA) and with systemic lupus erythematosus (SLE), compared with the degradation rate in degenerative joint disease and normal subjects. Carboxypeptidase activity was the same in all individuals, but ACE activity was increased in the RA and SLE patients. We examined the effects of aspirin, sodium salicylate, auranofin, penicillamine, and corticosteroids on kinin metabolism, and all of these were marked inhibitors of ACE; however, only penicillamine had any demonstrable inhibition of carboxypeptidase. These observations suggest rapid degradation of DBK in patients with untreated RA and SLE, whereas drugs utilized in therapy have the opposite effects. Studies to examine the role of DBK in disease manifestations are in progress. PMID- 3030336 TI - Hereditary amyloidosis: description of a new American kindred with late onset cardiomyopathy. Appalachian amyloid. AB - A family with hereditary amyloidosis characterized by peripheral neuropathy and cardiomyopathy is described. Lack of eye involvement sets their disease apart from the Indiana/Swiss familial amyloidotic polyneuropathy type II. The disease is of late onset; affected members die of cardiomyopathy after age 60. The late onset and lack of clinically significant neuropathy in several family members has led to misdiagnosis of the cardiomyopathy. Immunohistochemistry using antiprealbumin antiserum showed staining of amyloid deposits in nerve and heart. PMID- 3030337 TI - Drug-induced peripheral neuropathy in a patient with psoriatic arthritis. AB - A patient with psoriatic arthritis developed a peripheral neuropathy while taking hydroxychloroquine and naproxen. Although it was initially suspected that hydroxychloroquine was responsible for the neuropathy, subsequent rechallenge with naproxen demonstrated that clinical and electrophysiologic findings were related to routine pharmacologic doses of naproxen. PMID- 3030338 TI - Proglumide exhibits delta opioid agonist properties. AB - Recently, it was reported that proglumide, a cholecystokinin (CCK) antagonist, potentiates the analgetic effects of morphine and endogenous opioid peptides and reverses morphine tolerance by antagonizing the CCK system in the central nervous system of the rat. In order to provide additional insight into the mode of action of this agent, we assessed the effect of proglumide in the isolated guinea pig ileum and the mouse, rat and rabbit vas deferens. Furthermore, we studied the in vitro binding affinity of this substance to mouse brain synaptosomes. Our results show that proglumide inhibits, dose dependently, the electrically stimulated twitches in the mouse vas deferens and guinea pig ileum, but not in the rat or rabbit vas deferens. The inhibitory action of proglumide on the mouse vas deferens, but not on the guinea pig ileum, is antagonized by naloxone and by the selective delta-antagonist, ICI 174,864, in a competitive fashion. Other CCK antagonists were found to be devoid of such activity on the mouse vas deferens. In vitro binding studies showed that proglumide displaces D-ala-D-[leucine]5 enkephalin (DADLE), a delta agonist, but not ethylketocyclazocine (EKC), a preferentially selective kappa agonist. The effect of proglumide appeared to be elicited presynaptically since it did not alter the norepinephrine-induced contractions of the mouse vas deferens. Our results suggest that proglumide might exert its opiate-like effects by activation of delta-opioid receptors. PMID- 3030339 TI - The effects of N-allylnormetazocine on electric shock titration in squirrel monkeys. AB - The effects of the stereoisomers of the prototypic sigma agonist N allylnormetazocine were evaluated in squirrel monkeys trained to respond on an electric shock titration schedule. The monkeys responded on a fixed-ratio schedule to decrease the level of shock delivered to their tails. The effects of the isomers were compared to the effects of phencyclidine and morphine administered alone and also in combination with a rate-suppressing dose of morphine. Morphine increased the level at which shock was maintained without decreasing rates of responding in the presence of shock. Both the (+)-isomer and PCP produced increases in the levels at which the monkeys maintained the shock, but only in some animals. The (-)-isomer of N-allylnormetazocine resulted in either decreases or no effect on the level at which the monkeys maintained the shock. The isomers were equipotent in decreasing response rates. When tested in combination with morphine, only the (-)-isomer antagonized the shock-level increasing effects of morphine. Thus, the isomers had similar effects on response rates but the (+)-isomer, like PCP, was more effective in increasing the level at which the monkeys maintained electric shock, while only the (-)-isomer was effective as a morphine antagonist. These results suggest analgesic-like effects for the (+)-isomer of N-allylnormetazocine and substantiate the opiate-antagonist properties of the (-)-isomer. PMID- 3030340 TI - Epstein-Barr virus (EBV) and X-linked lymphoproliferative syndrome (XLP). AB - XLP was first described in 1975, when EBV was still focused on as an immediate oncogenic agent, but with some uncertainties raised by the absence of EBV in most non-endemic Burkitt lymphoma. The discovery of XLP refreshingly evidenced and popularized the concept, "EBV pathogenicity (oncogenicity) in immunocompromised hosts", which was later vastly substantiated by EBV-carrying lymphomas in organ transplanted and AIDS patients. Fatal (or severe) IM, acquired hypogammaglobulinemia (AH) and malignant lymphoma (MH) are 3 major phenotypes in XLP. Fatal IM occurs in 2/3 with a mortality rate of 85%. Lymphocyte infiltration (T cells and EBV-positive B cells) followed by their depletion with the appearance of macrophages. EBV-associated hemophagocytic syndrome in the bone marrow leads to hematocytopenia, ML, AH and ML with AH usually occur after EBV infection, but can occur before it. Reactivation pattern of EBV serology (high VCA, high EA, low EBNA), impaired generation of EBV-specific killer cell (poor regression), lowered NK activity, lowered 4/8 ratio and failure to mount IgG response to phi X174 have been recorded in XLP and carrier females. However, some or all of these are also found in non-XLP congenital immunodeficiencies as well as acquired immunodeficient states like advanced lymphoma or AIDS. In order to record XLP-specific defects, impaired help of autologous Ia-stimulated T4 cells or exaggerated suppression by T cells exposed to MA-pulsed macrophages is now being tested (Purtilo, personal communication). Fewer V region genes of T receptor may be a possibility. Restriction fragment length polymorphism (RFLP) by using probes from X chromosome may substantiate the precise genetics of this disease. PMID- 3030341 TI - Epstein-Barr virus infection and oncogenesis in primary immunodeficiency. AB - Lymphomas occurred in 3 of 16 Japanese patients with ataxia telangiectasia (AT) and Wiskott-Aldrich syndrome (WAS). The patients had a persistently reactivated Epstein-Barr virus (EBV) infection with a remarkable decrease in virus-specific cellular immunity. In these patients, the B lymphocytes were more sensitive to EBV-induced events and to cellular proto-oncogene activation than seen in the healthy counterparts. This immunologic and genetic background was considered to explain the massive lymphoproliferation in these primary immunodeficiency disorders. PMID- 3030342 TI - Unusual lymphoproliferation associated with chronic active Epstein-Barr virus infection. AB - A chronic active Epstein-Barr virus (EBV) infection with no significant underlying diseases was evidenced in 7 Japanese children and adolescents. In these patients, massive EBV-positive polyclonal lymphoproliferation occurred with extremely high EBV antibody titers. PMID- 3030343 TI - Impairment of T-cell control of Epstein-Barr virus infected B-cells in patients with adult T-cell leukemia. AB - The abilities of peripheral blood lymphocytes from 9 patients with adult T-cell leukemia (ATL) and 14 healthy control adults to cause regression of proliferating B-cell foci induced by Epstein-Barr virus (EBV) were examined. Marked regression of EBV-transformed B-cell foci was observed in lymphocytes from all 10 EBV seropositive donors but with none of those of the 4 EBV-seronegative donors in the control group. The lymphocyte of all 9 patients with ATL, who were EBV seropositive, caused little or no regression of B-cell foci like those of EBV negative healthy donors. These results suggest that the absence of regression of EBV-induced B-cell proliferation observed in ATL patients is due to impairment of EBV-specific killer T-cell activities. PMID- 3030344 TI - Epstein-Barr virus-enhancing plant promoters in east Africa. AB - The extracts of a certain African plant species, Euphorbia tirucalli, were found to have markedly enhancing effects on the activation of latent Epstein-Barr virus (EBV) genomes in the EBV carrying lymphoblastoid cells and also on EBV-induced transformation of human lymphocytes. The Euphorbia tirucalli was especially noticeable in highly endemic areas of Burkitt's lymphoma (BL), an EBV-associated malignancy, in Kenya and Tanzania. The activation of the latent EBV genome and the EBV-induced transformation enhancement were also observed with the soil and drinking water taken around the plants, strongly indicating that the people living in BL endemic areas are frequently exposed to such an EBV-enhancing plant promoter substance. PMID- 3030345 TI - Constitutive activation of oncogenes by chromosomal translocations in B-cell derived tumors. AB - The mechanisms of chromosomal translocations and its role in Burkitt lymphoma (BL), mouse plasmacytoma (MPC) and rat immunocytoma (RIC) are discussed with particular emphasis on the following questions: 1) Does the cis-relationship between the c-myc oncogene and one of the 3 Ig-loci play a causative role in the genesis of these tumors? 2) How does the juxtaposition activate the myc-gene? 3) What is the functional role of the translocation in the tumorigenic process? Question 1) can be answered with some certainty. In BL, the translocation has been found in 100% of cases so far, with no difference between endemic or nonendemic, EBV-carrying or EBV-negative cases. One exceptional line, BJAB, can be disregarded, since it is not a typical BL. In RIC, all examined tumor had the translocations so far. Only 90% of the MPCs carry the translocations, but high resolution banding of some translocation negative MPCs has shown that they carry an interstitial deletion in the D2/D3 region of Chr. 15, corresponding to the myc locus. Molecular analysis showed a complex rearrangement that has led to the juxtaposition of c-myc and IgH-switch sequences. Sequencing data indicated that they must have arisen by at least two independent translocations and one inversion. A similarly complex rearrangement was found in the first RIC that has been examined. The regularity of the association between the translocation events and the tumors where they occur, together with the similarities between the human, mouse and rat systems can be interpreted by postulating that the activation of c-myc by the translocation represents an essential step in the genesis of these tumors. 2) The transposed myc gene becomes constitutively activated. In all probability, this renders the gene resistant to cell cycle and differentiation dependent regulations that govern its expression in the normal chromosomal location. 3) The hypothesis is advanced that the translocation affects B-cells at the point where an antigen activated cell is about to leave the proliferative process, upon the waning of the antigenic stimulus, and enters a program towards a long lived memory cell. PMID- 3030346 TI - Substances affecting the infection and replication of human immunodeficiency virus (HIV). AB - The effects of various compounds were studied quantitatively on the growth of human immunodeficiency virus (HIV). For this we used a human T-cell leukemia virus type I carrying cell line, MT-4 which is most permissive for HIV infection. The results are summarized as follows: 1) Prostaglandin E2 and 12-0 tetradecanoylphorbol-13-acetate enhanced the production of HIV significantly in MT-4 cells as well as a continuous HIV producer Molt-4/HTLV-III cells. 2) Interferon gamma, retinoic acid and 3'-azido-3'-deoxythymidine inhibited the replication of HIV at the concentrations which were not cytotoxic. Mechanism of action of these compounds and its clinical implications are discussed. PMID- 3030347 TI - The Epstein-Barr virus genome and phenotypic expression during lytic cycle. AB - The Epstein-Barr Virus (EBV) has been studied extensively as a human cancer virus. Until recently, however, little was known about the viral genes encoding for different proteins involved in the virus immortalization and replication cycles. Most of the efforts have been directed at those genes expressed in immortalized cells. However, more recently, there has also been advances in the mapping of genes encoding for polypeptides expressed in the virus replication cycle and in the characterization of the proteins encoded by these genes. The purpose of this article is to review some of these new developments in identifying viral genes and their products. In addition, the current status of the development of a subviral vaccine against this virus is discussed. PMID- 3030348 TI - Epstein-Barr virus-specific cytoskeletal early antigen in EBV-transformed cell lines. AB - An Epstein-Barr virus (EBV)-specific determinant of early antigen (EA) character was found to be associated with intermediate filaments of EBV genome-activated cells. This determinant was defined with a murine monoclonal antibody (MAb) and with human MAb derived from lymphoblastoid cell lines of patients with acute infectious mononucleosis (IM). Such patients' antibodies either recognized intermediate filament determinants in general, or were restricted to the intermediate filament determinant of EBV genome-activated cells, as also recognized with the 1B6 murine MAb. The 1B6 determinant was a component of EA as defined by phosphonoacetate-resistance, methanol-sensitivity, and various co staining and antibody-blocking experiments. While anti-intermediate filament antibodies have been reported after various viral illnesses, the 1B6 and some patients' antibodies recognize an EBV-specific determinant which might function in the lytic cycle of these cells. PMID- 3030349 TI - Epstein-Barr virus seroepidemiology in China. AB - Since 1978 more than 300,000 sera from normal individuals were screened serologically in NPC high risk areas and prospective studies were carried out. Many patients were diagnosed in early stage. For example, in Wuzhou city, 20,726 persons over 40 years of age were screened; 1,138 persons were found to have IgA VCA antibody. Among them 18 NPC cases were detected; an additional 21 NPC patients were found within 5 year follow-up studies. Altogether there were 39 NPC patients. As compared to the patients in outpatient clinics, the frequency of NPC in stage I increased from 1.7% to 38.5% and in early stage (I + II) increased from 32% to 92.3%. IgA VCA antibody can be detected 5 years before the diagnosis of NPC in early stage was made. The detection rate of NPC from IgA VCA antibody positive persons is 38-374 times the incidence rate of NPC in the general population of the same age group. Follow-up studies on the change of IgA VCA antibody titer in antibody-positive and antibody-negative groups were also carried out for years. 10.9% of antibody-positive individuals became antibody negative and 5.4% seronegative persons converted to positive within 4 years. Eighty-eight per cent of NPC patients were detected in the group of no change of antibody titer or in the group of increasing antibody titer. No NPC patients were found in the original antibody negative group. PMID- 3030351 TI - Genomic expressions of human T-lymphotropic virus (HTLV-I). AB - Human T-lymphocyte cell line termed MT-2 is producing persistently HTLV-I virion and has a strong potential to transform human T-lymphocytes when cocultivated. The virion of HTLV-I (MT-2) was isolated and its RNA was extracted to analyze the gene and gene products of HTLV-I. HTLV (MT-2) virion RNA was translated in a rabbit reticulocyte lysate system in vitro in which a gag precursor polyprotein (p53) and a putative gag-prt fusion protein (p76) were synthesized from a full length 35S RNA. The full length provirus, HTLV-I (MT-2), was molecularly cloned and its genomic expression was examined transiently and permanently by transfecting in human lymphoid and non-lymphoid cells. The cloned provirus expressed the same virological activities as observed in naturally occurring infection of the virus. A new protease gene of HTLV-I was found and its function of the gene product was studied. PMID- 3030350 TI - Mechanism of the gene expression of HTLV-I and its association with ATL. AB - Human T-cell leukemia virus type 1 (HTLV-1) is the etiologic agent of adult T cell leukemia (ATL) and a trans-acting viral function was proposed to be involved in ATL development because of the non-specific provirus integration in leukemic cells and the frequent immortalization of helper T-cells by in vitro infection. An extra sequence "pX" in the HTLV-1 genome codes for three proteins, p40x-, p27x and p21x-, and the p40x- is trans-activator of transcription from the viral LTR. A sequence of 21 bp repeats in the LTR was found to be an enhancer and respond to the trans-activation by p40x-. The transient expression of p40x- also activates a cellular gene for interleukin 2 receptor (IL-2R) in helper T-cell lines. This induction of IL-2R may explain the mechanism of preferential growth of HTLV-1 infected cells and may be an early event of ATL development. For practical purposes, the env gene fragments was expressed in E. coli as fusion proteins with beta-galactosidase. Using these fusion proteins, a diagnostic system detecting anti-env antibodies was developed. Immunization of monkeys with these envelope fusion proteins protected the monkeys from the viral infection, suggesting possible usage of envelope proteins as vaccine. PMID- 3030353 TI - On the mechanism of the oscillatory activity of the brain. PMID- 3030352 TI - [Granular cell tumor of the gingiva. Report of a case and review of the terminological, clinical, histogenic and pathological criteria]. PMID- 3030354 TI - Catecholamines and aggression in animals. AB - This paper assesses the evidence for the role of catecholamines in the aggressive behaviour of animals. The effects of manipulating dopamine and noradrenaline function, either alone or in combination, are considered with respect to two categories of aggression, predatory and affective. Affective aggression is further subdivided into shock-induced defensive fighting, isolation-induced aggression and irritable aggression. The results indicate that catecholamines may not have a specific role in aggressive behaviour. Rather, they may act more to excite or inhibit general behavioural systems, although certain treatments do have a specific influence on aggressive behaviour. The review also highlights certain problems concerning the psychopharmacology of aggression; different species may make varying responses to the same treatment, whilst treatments exerting a similar pharmacological action may result in diverse behavioural effects. PMID- 3030355 TI - Aggression, defeat and opioid activation in mice: influences of social factors, size and territory. AB - The aggressive components and opioid-mediated behavioral consequences of various types of intraspecific agonistic interactions between individual male mice were examined. The size of the animals, their previous social history (group or isolation housing) and territory on which the encounter took place were varied to yield 26 different 'resident-intruder' paradigms. In these agonistic encounters the latency to first attack, number of bites and time to defeat, as well as the number of attack bouts present varied according to the 'resident-intruder' paradigm employed. The behavioral consequences of aggression and defeat, including analgesia, increased activity and augmented feeding were determined from the subordinate mice in 5 representative agonistic interactions. These behavioral responses, which had been previously shown to be mediated by endogenous opioid systems also varied according to the 'resident-intruder' paradigm employed. When both mice were group-housed, there was no agonistic behavior, regardless of the size of the mice or the testing arena. In isolated animals the defeat posture was only observed in 1 of the 19 paradigms. It is suggested that various 'resident-intruder' pairings and agonistic interactions can provide a reliable and useful means of examining differential naturalistic stress-induced endogenous opioid activation. PMID- 3030356 TI - Less-than-expected variability in evidence for three stages in memory formation. AB - Gibbs and Ng (1976, 1977) proposed a three-stage model of long-term memory formation on the basis of results from a one-trial-learning passive-avoidance procedure using chicks. Chisquare tests show that much of the evidence for this model (Gibbs & Ng, 1976, 1977, 1979, 1984a, 1984b), involving transient drops in retention and drug effects, is less variable than would be expected by chance. This should reduce belief in the model. PMID- 3030357 TI - Activation and expression of endogenous pain control mechanisms in rats given repeated nociceptive tests under the influence of naloxone. AB - In six experiments, it was found that animals administered the opiate receptor blocker naloxone prior to either hot-plate or tail-flick nociceptive tests developed reduced sensitivity to pain relative to animals tested under saline. The naloxone-induced analgesia was most pronounced following administration of 10 mg/kg naloxone, with weaker effects occurring at 0.5 and 2 mg/kg. The effect manifested itself in tests using mild (48.5 degrees hot-plate tests), but not more severe (52 degrees or 56 degrees hot-plate tests), intensities of nociceptive stimulation. The analgesia observed in animals tested under naloxone arose in part from the attenuation of the habituation of stress-induced analgesia produced by the novelty of the test apparatus, and in part from exposure to nociceptive stimulation. It appears to be mediated by a nonopiate mechanism; naloxone enhanced the analgesia produced by exposure to brief, continuous shock, but blocked the analgesia elicited by prolonged, intermittent shock (see Lewis, Cannon, & Liebeskind, 1980). We also found that administration of naloxone prior to nociceptive testing enhanced the development of conditioned autoanalgesia (as assessed by nociceptive tests conducted under saline), and that the enhanced conditioned autoanalgesia summated with the analgesic effect of morphine. The results are discussed in terms of the activation and expression of both opiate and nonopiate pain suppression mechanisms; their implications for models of situation specific morphine analgesic tolerance are discussed. PMID- 3030358 TI - Effects of alcohol on alpha-adrenergic receptor regulation of calcium ATPase in liver plasma membranes. AB - The effects of alpha 1- and alpha 2-adrenergic agonists, viz., phenylephrine and clonidine, respectively, were studied on rat liver plasma membrane Ca++-ATPase. Phenylephrine produced a 23% inhibition of enzyme activity at 5 microM. Prazosin, an alpha 1 antagonist, completely prevented the effect of phenylephrine. Clonidine produced a comparable inhibition of Ca++-ATPase, but was not reversed by the antagonist yohimbine, suggesting a lack of functionally significant alpha 2 receptors as previously reported. The results support the role of high-affinity Ca++-ATPase in liver plasma membranes in the control of cytosolic free Ca++ levels through regulation by alpha 1-adrenergic receptors. In vitro and acute ethanol exposure produced inhibition of plasma membrane Ca++-ATPase. In addition, ethanol treatment significantly reversed the inhibitory effect of phenylephrine on Ca++-ATPase. Chronic ethanol exposure for four weeks increased Ca++-ATPase activity over control and increased enzyme activity in the presence of phenylephrine. These results demonstrate that ethanol alters the alpha-adrenergic receptor interaction with Ca++-ATPase resulting in reduced receptor regulation of cytosolic Ca++ levels. These changes may prevent the liver from maintaining Ca++ levels for second messenger functions, such as glycolysis and gluconeogenesis. PMID- 3030359 TI - Depressed atrial inotropic response in the rat with chronic ethanol ingestion. AB - Consumption of ethanol for long periods of time has been correlated with cardiac dysfunction as well as changes in function of the autonomic nervous system. This study looked at the effect of chronic ethanol ingestion on atrial contractility and atrial muscarinic and beta adrenoreceptors. Male Long-Evans hooded rats were pair-fed on ethanol (E) or normal (N) liquid diet for 40 weeks. The E diet supplied 35-39% of calories as ethanol. The atria from E rats had significantly lower baseline and peak contractility. They also showed a higher incidence of failure induced by isoproterenol. There was no difference in concentration or binding characteristics of beta-adrenoreceptors or muscarinic receptors. The data suggest that the negative inotropism caused by ethanol ingestion is the result of some mechanism other than changes in autonomic receptors. PMID- 3030360 TI - Carbamate kinase of Lactobacillus buchneri NCDO110. I. Purification and properties. AB - Carbamate kinase has been prepared from Lactobacillus buchneri NCDO110. An approximately 91-fold increase in the specific activity of the enzyme was achieved. The purified extract exhibited a single band following polyacrylamide gel electrophoresis. The apparent molecular weight as determined by gel electrophoresis was about 97,000. The enzyme is stable for 2 weeks at -20 degrees C. Maximum enzymatic activity was observed at 30 degrees C and pH 5.4 in 0.1 M acetate buffer. L. buchneri carbamate kinase requires Mg2+ or Mn2+; its activity is higher with Mn2+. The activation energy of the reaction was 4078 cal mol-1 for the reaction with Mn2+ and 3059 cal mol-1 for the reaction with Mg2+. From a Dixon plot a pK value of 4.8 was calculated. The apparent Km values for ADP with Mg2+ or Mn2+ were 0.71 X 10(-3) and 1.17 X 10(-3) M, respectively, and the apparent Km values for carbamyl phosphate with Mg2+ or Mn2+ were 1.63 X 10(-3) and 1.53 X 10(-3) M, respectively. ATP and CTP acted as inhibitors of this reaction and the following values were obtained: Ki (ATP)Mg2+ = 9.4 mM, Ki (ATP)Mn2+ = 6.2 mM, and Ki (CTP)Mg2+ = 4.4 mM. PMID- 3030362 TI - [Biochemical aspects of lymphocyte metabolism. Immunomodulating role of cyclic nucleotides]. AB - In the present review lymphocyte metabolism has been examined with particular reference to ion and molecule flux variations through cell membrane. Furthermore, mitogenic stimulations causes a tremendous increase of cellular metabolism with remarkable modifications of oxidative processes and synthesis rate of enzymes and structural proteins. Finally, the role of cyclic nucleotides in the regulation of some lymphocyte functions has been analyzed. PMID- 3030361 TI - Idiotypic specificity of a human monoclonal anti-Rhesus(D) antibody. AB - A human monoclonal IgG anti-Rhesus(D) (H2D5D2) obtained after transformation of lymphoid cells by Epstein-Barr virus was purified from culture supernatant by affinity chromatography. Rabbits were immunized with the monoclonal anti-D. The rabbit antisera, after appropriate absorption, reacted only against the immunizing monoclonal anti-D. The antiidiotypic antibodies (anti-id ab) were purified and gave a complete inhibition of the monoclonal anti-D, whereas no inhibition was observed with monoclonal or polyclonal IgG or with sera containing high titres of anti-D. Polyclonal anti-D obtained from 118 immunized blood donors were coated on Rh-positive red cells. Agglutination by anti-id ab was observed in 4 cases (3.4%), indicating a cross-reactivity between the monoclonal anti-D and the polyclonal anti-D present in the sera of some immunized donors. PMID- 3030363 TI - Prevalence of HTLV III/LAV and other viral antibodies in normal and at risk populations. AB - The prevalence of HTLV III/LAV antibody has been studied in six groups of population: healthy subjects, a virological laboratory staff, prisoners, drug addicts, patients with lymphoadenopathy-associated syndrome (LAS) and patients with AIDS. Prevalences and levels in the same groups of HBsAg and CMV, HSV 1 and HSV 2 antibodies have been studied, too. As expected, the prevalence of HTLV III/LAV antibody increases in the six groups with the increasing of the risk while the prevalences of the other markers of viral infection do not show such a pattern. The levels of positivity of CMV antibody are significantly higher in the prisoners and LAS groups versus healthy subjects suggesting a possible role of this virus in the developing of full-blown AIDS. PMID- 3030364 TI - [Effect of mercaptans on serum IgA]. AB - Using reducing agents like dithiothreitol (DTT) 2.5 mM IgM lose their ability to bind antigens but they can be still detected by an anti-IgM antiserum. IgA are more resistant to reduction than IgM but using higher concentrations of DTT they lose both the ability to bind antigens and the possibility to be detected by an anti-IgA antiserum. Because of this fact IgA after mild reduction can result more or less inactivated according to the method employed to detect them. Using ELISA or similar methods that need an immunological detection of preformed immune complexes IgA-Ag, the loss of activity is more marked than using techniques such as HAI that detect them directly. PMID- 3030365 TI - The roles of cAMP and cAMP-dependent protein kinase in the regulation of adrenocortical functions: analysis using DNA-mediated gene transfer. AB - DNA-mediated gene transfer was used to evaluate the cause and effect relationship between mutations in cAMP-dependent protein kinase activity and cellular resistance of adrenocortical tumor cells to ACTH and cAMP. Protein kinase defective, Kin 8 adrenocortical tumor cells were transformed with genomic DNA from an ACTH- and cAMP-responsive adrenocortical cell line and screened for the recovery of morphological responses to the cAMP analog 8-bromo-cAMP (8BrcAMP). 8BrcAMP-responsive transformants were recovered with a frequency of approximately 0.5 per 10(3) transformation-competent cells. These transformants recovered the ability to round up in the presence of ACTH and were able to respond to both ACTH and 8BrcAMP with increased steroidogenesis. They also recovered cAMP-dependent protein kinase activity. The transformants, however, were unstable and concomitantly lost cAMP-dependent protein kinase activity and steroidogenic and morphological responses to ACTH and 8BrcAMP. These observations suggest that a single gene, probably the gene encoding the regulatory subunit of cAMP-dependent protein kinase, is responsible for the resistance of the Kin 8 mutant to ACTH and cAMP. PMID- 3030366 TI - Regulation of cAMP concentration by calmodulin-dependent cyclic nucleotide phosphodiesterase. AB - Bovine brain contains two major calmodulin (CaM) dependent phosphodiesterase isozymes which are homodimeric proteins with subunit molecular masses of 60 and 63 kilodaltons (kDa), respectively. The 60-kDa subunit isozyme can be phosphorylated by cAMP-dependent protein kinase, resulting in a decrease in the enzyme affinity towards CaM. The phosphorylation is blocked by Ca2+ and CaM and reversed by the CaM-stimulated phosphatase (calcineurin). The 63-kDa subunit isozymes can also be phosphorylated, but in this case by a CaM-dependent protein kinase(s). This phosphorylation is also accompanied by a decrease in the isozyme affinity towards CaM and can be reversed by the CaM-dependent phosphatase. Analysis of the complex regulatory properties of the phosphodiesterase isozymes has led to the suggestion that fluxes of cAMP and Ca2+ during cell activations are closely coupled and that the CaM-dependent phosphodiesterase isozymes play key roles in this signal coupling phenomenon. PMID- 3030367 TI - Translocation of Mg2+-dependent phosphatidate phosphohydrolase between cytosol and endoplasmic reticulum in a permanent cell line from human lung. AB - Incubation of A549 cells with digitonin for 4 min resulted in the release of over 90% of the lactate dehydrogenase activity into the medium. Approximately 80% of the Mg2+-dependent but only 7% of the Mg2+-independent phosphatidate phosphohydrolase activity was released in the presence of digitonin. Pretreatment of the cells with oleate reduced the efflux of the Mg2+-dependent phosphatidate phosphohydrolase activity to approximately 5% of total. Oleate did not affect the release of lactate dehydrogenase or the release of the Mg2+-independent phosphohydrolase activity. Incubation of A549 cells with [3H]oleate for 60 min led to incorporation of the label into phosphatidic acid, phosphatidylethanolamine, phosphatidylcholine, diacylglycerol, monoacylglycerol, and triacylglycerol, in ascending order. When the level of exogenous oleate was increased to over 2.0 mM, there was a marked increase in the incorporation into monoacylglycerol and diacylglycerol. Only small amounts of radioactivity were associated with phosphatidic acid. Time course studies revealed that the amount of radioactive phosphatidate remained low throughout the incubation period. These investigations were interpreted to indicate that free fatty acids can promote the translocation of the Mg2+-dependent phosphatidate phosphohydrolase activity from cytosol to membrane fractions. This translocation could, at least theoretically, function to facilitate the metabolism of increased amounts of phosphatidate. PMID- 3030368 TI - L-malate transport and proton symport in vesicles prepared from Pseudomonas putida. AB - In vesicles from glucose-grown Pseudomonas putida, L-malate is transported by nonspecific physical diffusion. L-Malate also acts as an electron donor and generates a proton motive force (delta p) of 129 mV which is composed of a membrane potential (delta psi) of 60 mV and a delta pH of 69 mV. In contrast, vesicles from succinate-grown cells transport L-malate by a carrier-mediated system with a Km value of 14.3 mM and a Vmax of 313 nmol X mg protein-1 X min-1, generate no delta psi, delta pH, or delta p when L-malate is the electron donor, and produce an extravesicular alkaline pH during the transport of L-malate. A kinetic analysis of this L-malate-induced proton transport gives a Km value of 16 mM and a Vmax of 667 nmol H+ X mg protein-1 X min-1. This corresponds to a H+/L malate ratio of 2.1. The failure to generate a delta p in these vesicles is considered, therefore, to be consistent with the induction in succinate-grown cells of an electrogenic proton symport L-malate transport system. PMID- 3030369 TI - Role of phospholipid fatty acids on the kinetics of high and low affinity sites of cytochrome c oxidase. AB - The nature of the interactions between cytochrome c oxidase and the phospholipids in mitochondrial membranes has been investigated by varying the nature of the fatty acyl components of Saccharomyces cerevisiae. A double fatty acid yeast mutant, FAI-4C, grown in combinations of unsaturated (oleic, linoleic, linolenic, and eicosenoic) and saturated (lauric and palmitic) fatty acids, was employed to modify mitochondrial membranes. The supplemented fatty acids constituted a unique combination of different acyl chain lengths with varying degrees of unsaturation which were subsequently incorporated into mitochondrial phospholipids. Phosphatidylethanolamine and cardiolipin, the predominant phospholipids of the inner mitochondrial membrane, were characterized by their high levels of supplemented unsaturated fatty acids. Increasing the chain length or the degree of unsaturation of mitochondrial membrane phospholipids had no effect on altering the nature of the phospholipid polar head group but did result in a profound change on the specific activity of cytochrome c oxidase. When studied under conditions of different ionic strengths and pHs the enzyme's activity, as documented by Eadie-Hofstee plots, showed biphasic kinetics. The kinetic parameters for the low affinity reaction were greatly influenced by the changes in the membrane fatty acids and only marginal effects were noted at the high affinity reaction site. The discontinuities in the steady-state fluorescence anisotropy of 1,6-diphenyl-1,3,5-hexatriene, monitored at increasing temperatures, suggested that changes in membrane fluidity were conditioned by alterations in mitochondrial membrane fatty acid constituents. These results indicate that the lipid changes affecting the low affinity binding site of cytochrome c oxidase may be the result of lipid-protein interactions which lead to enzyme conformational changes or may be due to gross changes in membrane fluidity. It may, therefore, follow that this enzyme site may be embedded in or be juxtaposed to the outer surface of the inner mitochondrial membrane bilayer in contrast to the high affinity site which has been shown to be significantly above the membrane plane. PMID- 3030370 TI - Cannabinoids reduce fertility of sea urchin sperm. AB - Cannabinoids are potent pharmacological substances derived from marihuana. The effects of delta 9-tetrahydrocannabinol (THC), cannabinol (CBN), and cannabidiol (CBD) on fertilization in the sea urchin Strongylocentrotus purpuratus were investigated. Insemination of THC-treated eggs (5-400 microM) with excess sperm did not result in polyspermic fertilization. At minimal sperm densities, THC (0.1 10 microM) inhibited fertilization in a dose-dependent manner. Pretreatment of eggs with THC did not reduce their receptivity to sperm. Pretreatment of sperm with THC reduced their fertilizing capacity. The concentration of THC required to reduce sperm fertility by 50% was 1.1 +/- 1.1 microM. The fertilizing capacity of THC-treated sperm depended on concentration of sperm and duration of pretreatment. The fertility of sperm at minimal densities was reduced by 50% at 129.3 +/- 43 s treatment with 10 microM THC. The adverse effect of THC on sperm fertility was reversible. CBN and CBD at comparable concentrations (0.1-10 microM) inhibited fertilization in a manner similar to THC. First division was not delayed in zygotes that were fertilized with sperm pretreated with 10 microM THC. These studies show that cannabinoids directly affect the process of fertilization in sea urchins by reducing the fertilizing capacity of sperm. PMID- 3030371 TI - Saturation analysis of cellular retinoid binding proteins: application to retinoic acid resistant human neuroblastoma cells and to human tumors. AB - A method for saturation analysis of cellular retinoic acid and retinol binding proteins, CRABP and CRBP, respectively, in cultured cells and human tumor samples, and its application to a retinoic acid resistant subline of the human neuroblastoma LA-N-5 cell line is described. Assessment of retinoid binding was accomplished by incubation of cytosols with increasing concentrations of [3H]retinoid (28-43 Ci/mmol; 1 Ci = 37 GBq) for 24 h. Bound retinoid was separated from free retinoid by adsorption with dextran-coated charcoal. Nonspecific binding was quantitated in parallel incubations which had been treated with p-chloromercuribenzene sulfonate (PCMBS), resulting in selective elimination of sulfhydryl-dependent ligand binding to both CRABP and CRBP. Quantitation was accomplished by Scatchard analysis of specific (PCMBS sensitive) binding. Employing this technique, specific retinoid binding was attributed to the presence of 2S macromolecules which displayed the known properties of CRABP and CRBP, namely ligand specificity, saturability, high ligand affinity, and PCMBS sensitivity. The apparent dissociation constants (Kd) for retinoic acid binding in cytosols prepared from murine 3T6 fibroblasts, rat testes, and a human ovarian tumor were 7, 11, and 35 nM, respectively. These preparations also bound retinol with high affinity, exhibiting Kds of 12, 26, and 48 nM, respectively. A retinoic acid resistant subline of LA-N-5 cells designated LA-N-5-R9 was established by long-term culture in the presence of 10(-6) M retinoic acid. This subline is resistant to the effects of retinoic acid in that it requires a 10 fold higher concentration of retinoic acid for 50% inhibition of growth than the parent line and displays no retinoic acid induced morphologic differentiation. Saturation analysis of CRABP in the parent and resistant subline reveal no significant alteration in either CRABP content or affinity. These results indicate that resistance to retinoic acid induced differentiation in LA-N-5-R9 occurs distal to CRABP binding or that CRABP does not mediate this response to retinoic acid. PMID- 3030372 TI - Regulation of the number of Na+,K+-pump sites after mitogenic activation of lymphocytes. AB - The early activation of Na+,K+-ATPase-mediated ion fluxes after concanavalin A (ConA) stimulation of pig lymphocytes is caused by an increase in intracellular Na+ concentration. A second mechanism of regulation of Na+,K+-ATPase activity becomes apparent between 3 and 5 h after mitogenic stimulation, but prior to onset of increase in cell volume; this consists of an increase (about 75%) in the number of ouabain-binding sites (from 35 X 10(3) +/- 12 X 10(3)/cell in resting to 60 X 10(3) +/- 27 X 10(3)/cell in activated lymphocytes). The increase in ouabain binding was attributed to an increase in the number of active Na+,K+ ATPase molecules, based on the following evidence: there was an increase in the Vmax of ouabain binding, without variation in the Km; the increase in ouabain binding was accompanied by a proportional increase in K+ influx, when the assay was performed in the presence of the Na+ ionophore monesin, which was used to eliminate the difference in intracellular Na+ concentration between resting and activated cells; there was proportionality between ouabain-inhibitable ATPase activity in permeabilized cells and the number of ouabain-binding sites in resting and activated lymphocytes. The ConA-induced increase in ouabain-binding sites was influenced neither by amiloride nor by incubation in low Na+ medium, under conditions which prevented both increase in intracellular Na+ concentration and K+ influx. Increase in intracellular Na+ concentration was ineffective in altering the number of active pump molecules in resting cells. During incubation with ConA, the presence of ouabain did not affect the increase in ouabain-binding sites; thus, regulation of the number of pump sites is independent of the regulation of their activity. The ConA-induced increase in number of ouabain binding sites did not require protein synthesis; indeed, cycloheximide, anisomycin, and puromycin, under conditions in which they inhibited protein synthesis by by 95%, induced the increase to approximately the same extent as did ConA. This suggests the presence in resting lymphocytes of a rapidly turning over protein that either prevents the ATPase subunits from assembling or from integrating into the membrane. PMID- 3030373 TI - [Chemotaxic activity of granulocytes in toluene diisocyanate-induced asthma]. PMID- 3030374 TI - [Preliminary study of Na/K-ATPase activity in erythrocyte ghosts in pregnancy induced hypertension]. PMID- 3030375 TI - [ESR study of metabolic characteristics of erythrocytes in the first hours of life]. PMID- 3030377 TI - [Muscle pathology in mitochondrial myopathy]. PMID- 3030376 TI - [Effect of cocaine administration on experimental hepatitis in the mouse caused by MHV-3 virus: preliminary results]. PMID- 3030378 TI - [Neurological approach to mitochondrial abnormalities]. PMID- 3030379 TI - [Clinical and biochemical approach to mitochondrial cytopathy--defects of the respiratory chain]. PMID- 3030381 TI - Mechanism of protein translocation across the endoplasmic reticulum membrane. PMID- 3030380 TI - Nonmuscle actin-binding proteins. PMID- 3030383 TI - Management of the mobile fibrous ridge in the atrophic maxilla using porous hydroxyapatite blocks: a preliminary report. PMID- 3030382 TI - [Recent drug therapy of senile dementia]. PMID- 3030384 TI - Recovery of respiration after neuromuscular blockade with alcuronium. AB - Alcuronium 0.2 mg kg-1 was given to six patients to investigate the simultaneous recovery of breathing and peripheral neuromuscular function. Anaesthesia was maintained with 66% nitrous oxide in oxygen supplemented with 0.5% halothane, and the patients were ventilated to normocarbia. Patients were disconnected from the ventilator after the reappearance of the tetanic response. This response returned at a mean time of 19.2 min after the injection of alcuronium and oxygenation was maintained thereafter by means of apnoeic diffusion. Spontaneous breathing returned at a mean time of 23.6 min after the injection of alcuronium. Sixty minutes after the administration of alcuronium, respiratory exchange was judged adequate, and at that time neuromuscular function was still markedly depressed with a tetanic height less than 25% of control. It was concluded that, because of the slow recovery of neuromuscular function, alcuronium should be reserved for the longer surgical procedure. PMID- 3030385 TI - In vivo release of cyanide from sodium nitroprusside. AB - Brain cytochrome oxidase activity was measured after the in vitro addition of potassium cyanide (KCN) or sodium nitroprusside (SNP). Activity of cytochrome oxidase was sensitive to KCN; however, this activity was unaffected by SNP. In SNP- and KCN-treated animals brain cytochrome oxidase activities were measured. At 3 min after SNP injection, inhibition of the enzymatic activity was the same as 1 min after KCN injection. Time to death for SNP-treated animals was longer than for KCN-treated animals. These data suggest that cyanide was released in vivo from SNP and that time was necessary for this release to occur. PMID- 3030388 TI - Metabolism of [14C]arachidonic acid by polymorphonuclear leukocytes in patients with psoriasis. AB - The formation of LTB4 and its omega-oxidation products 20-hydroxy- and 20-carboxy LTB4 from exogenous [14C]arachidonic acid (AA) by neutrophils from 12 psoriatic patients and 10 healthy controls was investigated. Only a slight difference was detected in the mean amount of [14C]LTB4 produced. In contrast, the amounts of [14C]omega-oxidation products obtained from psoriatic PMN were 2.4-fold higher than the amounts from PMN of healthy controls. We conclude that in vitro, psoriatic PMN synthesize more LTB4 from exogenous AA than do PMN of healthy individuals and due to an efficient omega-oxidation system, the net release of LTB4 in both groups appears to be similar. PMID- 3030386 TI - Antihypertensive effect of single doses of enalapril in hypertensive patients treated with bendrofluazide. AB - This study was designed to investigate the validity of the then current recommendations for initiation of therapy with enalapril in hypertensive patients on treatment with a diuretic. Enalapril in single doses of 10 and 20 mg was given to 13 hypertensive patients on treatment with bendrofluazide 5 mg daily in a randomised, crossover, placebo controlled study. The mean maximal reduction in blood pressure was similar with both doses (35/20 mmHg supine, 38/20 mmHg standing), occurred on average within 6 h of tablet ingestion, and was not accompanied by any significant change in heart rate. Three patients experienced symptomatic hypotension. In one patient this was incapacitating after 10 mg and precluded exposure to 20 mg. This study shows that in hypertensive patients receiving treatment with diuretics, the addition of enalapril should be undertaken with caution. An optimal starting dose of enalapril in such patients remains to be confirmed. PMID- 3030390 TI - Immunoreactive prolyl hydroxylase in patients with primary and secondary myelofibrosis. AB - Prolyl hydroxylase (PH) is an important enzyme in collagen synthesis. It is required for the hydroxylation of prolyl residues in peptide chains in collagen synthesis. Serum PH activity was measured in patients with primary myelofibrosis (agnogenic myeloid metaplasia and myelofibrosis with prior history of polycythaemia vera), in patients with secondary myelofibrosis (in association with carcinoma metastasis), in patients with other myeloproliferative disorders and in controls (anaemia patients and normal volunteers). Both primary and secondary myelofibrosis had significantly elevated PH values, while other myeloproliferative disorders did not differ from normal controls. Increased PH levels in patients with primary and secondary myelofibrosis may signify increased collagen synthesis and may serve as an indicator for the development of fibrosis in the course of myeloproliferative disease. PMID- 3030389 TI - Enzyme cytochemistry of neutrophil granulocytes. PMID- 3030387 TI - Haemodynamic and humoral effects of oral perindopril, an angiotensin converting enzyme inhibitor, in man. AB - The tolerance to and dynamic effects of 1 week's oral treatment with the angiotensin converting enzyme inhibitor, perindopril, were assessed in a placebo controlled, parallel group study in 36 normotensive males. The daily dose of perindopril was 1, 2, 4, 8 or 16 mg. The drug was well tolerated and produced no change in routine haematology or serum biochemistry tests. Dose related inhibition of plasma angiotensin converting enzyme was observed. Perindopril 16 mg produced 90% inhibition 4 h after dosing and 60% after 24 h. A dose related rise in plasma renin activity followed doses of 4 mg and over. The renin remained above the normal range for 24 h. Perindopril caused a modest lowering of plasma aldosterone levels but had no effect on plasma adrenaline or noradrenaline levels. Standing diastolic blood pressure was lowered, particularly with 16 mg daily of perindopril but only a slight rise in heart rate occurred. Perindopril appears to be a well tolerated inhibitor of plasma angiotensin converting enzyme, with predictable effects on the renin angiotensin system and blood pressure. An appropriate dose range for further study would appear to be 4 to 16 mg daily. PMID- 3030391 TI - Enhanced lipid peroxidation and lysosomal enzyme activity in the lungs of rats with prolonged pulmonary deposition of crocidolite asbestos. AB - The interaction of UICC crocidolite asbestos with biological membranes in vivo was studied in rats after a single intratracheal dose of a suspension of 20 mg of fibres per rat. Development of lung fibrosis (increased level of hydroxyproline, a collagen index together with corresponding pathomorphological alteration) confirmed the penetration of crocidolite fibres into the lungs in the course of seven months exposure. The pulmonary deposition of crocidolite affected the lysosomal membranes of lung cells as manifested by (1) enhanced lipid peroxidation with (2) stimulation (release) of activity of beta-glucuronidase and cathepsin D. Enhanced lipid peroxidation and activity of beta-glucuronidase may contribute to the delayed, carcinogenic effects of crocidolite asbestos. PMID- 3030392 TI - Cyproterone acetate, an alternative progestogen in postmenopausal hormone replacement therapy? Effects on serum lipids and lipoproteins. AB - Serum lipids and lipoproteins were studied in 76 healthy postmenopausal women treated for 1 year with either oestradiol valerate sequentially combined with the anti-androgenic progestogen cyproterone acetate (CPA) or placebo. The women were examined every 3 months between days 18 and 21 of the tablet cycle, where the progestogen had been added to the oestrogen for at least 6 days. Combined oestrogen-CPA therapy resulted in significantly reduced levels of total serum cholesterol and LDL-cholesterol at all examinations, but serum triglycerides and HDL-cholesterol levels were similar in the two groups. Total serum cholesterol and LDL-cholesterol were reduced by approximately 5% (P less than 0.01) and 8% (P less than 0.01), respectively, after 1 year of combined oestrogen-CPA therapy in comparison with both the initial values and the placebo group but serum triglycerides and HDL-cholesterol levels were unchanged in both groups. PMID- 3030393 TI - Construction and characterization of an active factor VIII variant lacking the central one-third of the molecule. AB - The primary structure of factor VIII consists of 2332 amino acids that exhibit 3 distinct structural domains, including a triplicated region (A domains), a unique region of 909 amino acids (B domain), and a carboxy-terminal duplicated region (C domains), that are arranged in the order A1-A2-B-A3-C1-C2. The B domain (residues 741-1648) of factor VIII is lost when factor VIII is activated by thrombin, which proteolytically processes factor VIII to active subunits of Mr 50,000 (domain A1), 43,000 (domain A2), and 73,000 (domains A3-C1-C2). To determine if the B domain is required for factor VIII coagulant activity, a variant was constructed by using recombinant DNA techniques in which residues 797-1562 were eliminated. This shortened the B domain from 909 to 142 amino acids. This variant factor VIIIdes-797-1652 was expressed in mammalian cells and was found to be functional. The factor VIIIdes-797-1562 protein was purified and shown to be processed by thrombin in the same manner as full-length factor VIII. The factor VIIIdes-797 1562 variant also bound to von Willebrand factor (vWF) immobilized on Sepharose. These results indicate that most of the highly glycosylated B domain of factor VIII is not required for the expression of factor VIII coagulant activity and its interaction with vWF. PMID- 3030394 TI - Domain organization of chicken gizzard myosin light chain kinase deduced from a cloned cDNA. AB - Myosin light chain kinases (MLCK) are the most studied of the calmodulin activated enzymes; however, minimal sequence information is available for the smooth muscle form of the enzyme. The production of an antibody against the enzyme and the use of expression vectors for constructing cDNA libraries have facilitated the isolation of a cDNA for this kinase. The derived amino sequence was found to contain a region of high homology (54%) to the rabbit skeletal muscle enzyme and also very significant homology (35%) to the catalytic subunit of phosphorylase b kinase and cGMP-dependent protein kinase. All of these homologies were found in the known catalytic domains of these enzyme, thus enabling us to predict the location of the catalytic domain for the chicken gizzard myosin light chain kinase. Within the catalytic domain a consensus sequence for an ATP-binding site was located. Subcloning and expression of different regions of the cDNA defined a 192 base pair fragment coding for the calmodulin-binding domain of MLCK. Both of the cAMP-dependent protein kinase phosphorylation sites were identified by sequence homology. A linear model for MLCK is presented placing the various domains in relative position. Northern blot analysis and S1 protection and mapping experiments have revealed that the mRNA for MLCK is 5.5 kilobases in length, but there also exists a second mRNA of 2.7 kilobases that shares a high degree of homology with about 520 base pairs at the 3' end of the cDNA for MLCK. PMID- 3030395 TI - Purification and characterization of high Ca2+-requiring neutral proteases from porcine and bovine brains. AB - Porcine and bovine brain high Ca2+-requiring neutral proteases were purified to homogeneity by the same isolation procedures, and their properties were compared. A high degree of similarity existed between the two proteases. The purification procedures included ion-exchange chromatography on DEAE-cellulose, hydrophobic chromatography on phenyl-Sepharose CL-4B, second DEAE-cellulose chromatography, second phenyl-Sepharose CL-4B chromatography, and gel filtration on Ultrogel AcA 34. Both purified enzymes were composed of Mr 75,000 and 29,000 subunits, as shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Both enzymes required 250 microM Ca2+ for half-maximal activity and 700 microM Ca2+ for maximal activity. Sr2+ and Ba2+, but not Mg2+ or Mn2+, also activated both enzymes but not as effectively as Ca2+. Both enzymes displayed maximum activity at pH 7.5-8.0. Leupeptin, antipain, and trans-epoxysuccinyl-L-leucylagmatine inhibited both enzymes. Neurofilament triplet proteins and microtubule-associated proteins were extensively hydrolyzed by both proteases, but tubulin and actin were not hydrolyzed. The amino acid compositions of the two proteases were very similar. Antisera against bovine brain protease cross-reacted with porcine brain protease when examined by immunoelectrotransfer blot techniques. PMID- 3030396 TI - Partial amino acid sequence of human thrombospondin as determined by analysis of cDNA clones: homology to malarial circumsporozoite proteins. AB - A lambda gt 11 library prepared from human umbilical vein endothelial cell RNA was screened for cDNAs encoding thrombospondin. Reagents included a monospecific antibody to human thrombospondin and a mixture of four synthetic oligodeoxyribonucleotides derived from an amino acid sequence near the NH2 terminus of mature human thrombospondin. Two series of cDNA clones coding for sequences at the 5' and 3' ends of thrombospondin mRNA, respectively, were isolated. The nucleotide sequence of a 1.3-kilobase (kb) 5' clone (lambda TS-33) coded for 99 bases of 5' untranslated RNA, a signal peptide of 18 amino acids, and the first 379 amino acids of thrombospondin. Northern blot analysis with lambda TS-33 detected a single mRNA species of approximately 6.0 kb in rat aortic smooth muscle cell RNA. Thrombospondin mRNA levels increased rapidly, but transiently, in quiescent smooth muscle cells treated with platelet-derived growth factor. The kinetics of this response were very similar to those of the thrombospondin protein to this growth factor. There was significant homology in amino acid sequence between thrombospondin and a conserved region in the circumsporozoite protein of two malarial sporozoites. This region of thrombospondin may therefore represent a potential recognition site for a cell surface thrombospondin receptor. PMID- 3030397 TI - Construction and characterization of a site-directed CC-1065-N3-adenine adduct within a 117 base pair DNA restriction fragment. AB - The design, construction, and characterization of a site-directed CC-1065-N3 adenine adduct in a 117 base pair segment of M13mpI DNA are described. CC-1065 is an extremely potent antitumor antibiotic produced by Streptomyces zelensis. Previous studies have demonstrated that the cyclopropyl ring of CC-1065 reacts quite specifically with N3 of adenine in double-stranded DNA to form a CC-1065 DNA adduct. Following alkylation, the drug molecule lies snugly within the minor groove of DNA, overlapping with five base pairs for which a marked sequence preference exists [Hurley, L. H., Reynolds, V. R., Swenson, D. H., Petzold, G. L., & Scahill, T. A. (1984) Science (Washington, D.C.) 226, 843-844]. On the basis of the unique characteristics of the reaction of CC-1065 with DNA and the structure of the resulting DNA adduct, we have designed a general strategy to construct a site-directed CC-1065-DNA adduct in a restriction fragment. The presence of unique AluI and HaeIII restriction enzymes sites on each side of a high-affinity CC-1065 binding sequence (5'-GATTA) permitted the preparation of a partial duplex DNA molecule containing the CC-1065 binding sequence in the duplex DNA region. Since CC-1065 only binds to duplex DNA, potential CC-1065 binding sequences in the long single-stranded regions were protected from drug binding during the construction process.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3030398 TI - Atrial natriuretic factor receptor on cultured Leydig tumor cells: ligand binding and photoaffinity labeling. AB - Atrial natriuretic factor (ANF) is a peptide hormone discovered recently from the heart atrium that possesses potent natriuretic and vasorelaxant activities. Recently we found that ANF markedly stimulates intracellular cGMP and almost completely inhibits cAMP accumulation in testicular interstitial tumor cells [Pandey, K. N., Kovacs, W. J., & Inagami, T. (1985) Biochem. Biophys. Res. Commun. 133, 800-806]. These actions of ANF suggest the presence of ANF receptors in testicular interstitial cells. In this study, cultured murine Leydig tumor cells have been shown to contain specific binding sites for ANF. Saturation binding studies indicated a single class of binding sites with a Kd of 5 X 10(-9) M at a density of 2 X 10(6) sites/cell. The binding of mono[125I]iodo-ANF (125I ANF) was competed by unlabeled ANF in a dose-dependent manner. Hormones unrelated to ANF such as angiotensin I, bovine luteinizing hormone, and human chorionic gonadotropin were not able to compete against 125I-ANF. The binding of 125I-ANF was rapid, reaching maximum levels in 15 min at 4 degrees C. At 37 degrees C, the cell-bound 125I label was quickly decreased. Pretreatment of cells with NH4Cl, chloroquine, or NaN3 resulted in significant increases in maximum levels of the cell-bound 125I radioactivity. A photoaffinity reagent for ANF receptor was prepared by reacting ANF with succinimido 4-azidobenzoate, and resultant 4 azidobenzoyl- (AZB-) ANF was purified by high-performance liquid chromatography (HPLC). AZB-ANF was radioiodinated by use of chloramine T and purified again by HPLC.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3030399 TI - Structure-activity analysis of the activation of pertussis toxin. AB - Bordetella pertussis, the causative agent of whooping cough, releases pertussis toxin in an inactive form. The toxin consists of an A protomer containing one S1 peptide subunit and a B oligomer containing several other peptide subunits. The toxin binds to cells via the B oligomer, and the S1 subunit is activated and expresses ADP-ribosyltransferase and NAD glycohydrolase activities. Treatment of purified toxin with dithiothreitol (DTT) in vitro increases both activities. ATP and the detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS) synergistically reduce the A0.5 (activation constant) for DTT from greater than 100 mM to 200 microM. We studied the structure-activity relationships of activators of the toxin. In the presence of CHAPS (1%) and DTT (10 mM) the following compounds increased the NAD glycohydrolase activity of the toxin with the following A0.5's in microM and fraction of the ATP effect in parentheses: ATP, 0.2 (1.0); ADP, 6 (0.8); UTP, 15 (0.7); GTP, 35 (0.6); pyrophosphate, 45 (0.7); triphosphate, 60 (0.6); tetraphosphate, greater than or equal to 170 (greater than or equal to 0.4). Thus, the polyphosphate moiety is sufficient to stimulate the toxin, and the adenosine moiety confers upon ATP its extraordinary affinity for the toxin. Phospholipid and detergents could substitute for CHAPS in the activation of the toxin. Glutathione substituted for DTT with an A0.5 of 2 mM, a concentration within the range found in eucaryotic cells. Thus, membrane lipids and cellular concentrations of glutathione and ATP are sufficient to activate pertussis toxin without the need for a eucaryotic enzymatic process. PMID- 3030400 TI - Lipid mobility and order in bovine rod outer segment disk membranes. A spin-label study of lipid-protein interactions. AB - Lipid-protein interactions in bovine rod outer segment disk membranes have been studied by using a series of eight stearic acid spin-label probes which were labeled at different carbon atom positions in the chain. In randomly oriented membrane dispersions, the electron spin resonance (ESR) spectra of the C-8, C-9, C-10, C-11, C-12, C-13, and C-14 atom positional isomers all apparently consist of two components. One of the components corresponds closely to the spectra obtained from dispersions of the extracted membrane lipids, and the other, which is characterized by a considerably greater degree of motional restriction of the lipid chains, is induced by the presence of the protein. Digital subtraction has been used to separate the two components. The proportion of the motionally restricted lipid component is approximately constant, independent of the position of the spin-label group, and corresponds to 30-40% of the total spin-label spectral intensity. The hyperfine splitting of the outer maxima in the difference spectra of the motionally restricted component decreases, and concomitantly, the line widths increase with increasing temperature but change relatively little with increasing distance of the spin-label group from the polar head-group region. This indicates that the corresponding chain motions of the protein interacting lipids lie in the slow-motion regime of spin-label ESR spectroscopy (tau R approximately 10(-8) S) and that the mobility of these lipids increases with increasing temperature but does not vary greatly along the length of the chain. The data from the hyperfine splittings also suggest the existence of a polarity gradient immediately adjacent to the protein surface, as observed in the fluid lipid regions of the membrane. The more fluid lipid component is only slightly perturbed relative to the lipids alone (for label positions 5-14, inclusive), indicating the presence of chain motions on the nanosecond time scale, and the spectra also reveal a similar polarity profile in both lipid and membrane environments. ESR spectra have also been obtained as a function of magnetic field orientation with oriented membrane samples. For the C-14 atom positional isomer, the motionally restricted component is observed to have a large hyperfine splitting, with the magnetic field oriented both parallel and perpendicular to the membrane normal. This indicates that the motionally restricted lipid chains have a broad distribution of orientations at this label position.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3030401 TI - Competition for formation of nucleosomes on fragmented SV40 DNA: a hyperstable nucleosome forms on the termination region. AB - We have studied the relative abilities of different simian virus 40 (SV40) DNA segments to reconstitute into nucleosomes in vitro. The SV40 genome was separated into 15 discrete fragments by restriction endonuclease digestion and reconstituted with calf thymus core histones under conditions of varying histone to-DNA ratios. Three fragments show very different abilities to form nucleosomes when low histone-to-DNA ratios require all fragments to compete for available histones. Two of these fragments, both from within protein-coding regions, are significantly underreconstituted. The third fragment, covering the SV40 termination region, competes much more effectively for histones than the other 14 fragments. The fragment containing the SV40 origin region formed nucleosomes with about average probability. Overall, the SV40 fragments differed by approximately an order of magnitude in their abilities to support nucleosome formation in vitro. The stability of the nucleosomes was measured by challenge with high concentrations of the destabilizing reagent heparin. The fragment that reconstituted most effectively also formed nucleosomes that were unusually stable to heparin challenge. These observations are intriguing since this fragment contains the sequences where replication of SV40 DNA commonly terminates and where early messenger RNA synthesis may terminate as well. The existence of unique hyperstable nucleosomes in this region suggests the interesting possibility that such nucleosomes may assist in termination events by assisting in the pausing of replication or transcription complexes. PMID- 3030403 TI - Parallax method for direct measurement of membrane penetration depth utilizing fluorescence quenching by spin-labeled phospholipids. AB - This report describes a method suitable for determining the depth of a wide variety of fluorescent molecules embedded in membranes. The method involves determination of the parallax in the apparent location of fluorophores detected when quenching by phospholipids spin-labeled at two different depths is compared. By use of straightforward algebraic expressions, the method allows calculation of depth in angstroms. Furthermore, the analysis can be extended to quenching by energy-transfer acceptors or brominated probes under appropriate conditions. Application of the method to quenching of 7-nitro-2,1,3-benzoxadiazol-4-yl (NBD) labeled lipids by spin-labeled lipids located at three different depths is demonstrated in model membranes. It is shown that the calculated depths of the NBD groups are self-consistent to the extent that they are the same no matter which two spin-labels have been used in a particular experiment. In addition, the calculated depth is independent of spin-label concentration in the membrane within +/- 1 A, ruling out major effects due to spin-label perturbation. The quenching experiments show that the location of the NBD group in head-group labeled phosphatidylethanolamine is at the polar/hydrocarbon interface and that of an NBD label on the "tail" of cholesterol is deeply buried, as expected. Unexpectedly, NBD labels placed at the end of fatty acyl chains of phosphatidylcholines are also near the polar/hydrocarbon interface. Presumably, the polarity of the NBD group results in "looping" back to the surface of the NBD groups attached to flexible acyl chains. PMID- 3030402 TI - Resolution of conformational states of spin-labeled myosin during steady-state ATP hydrolysis. AB - We have measured the conventional electron paramagnetic resonance (EPR) spectrum of spin-labeled myosin filaments as a function of the nucleotide occupancy of the active site of the enzyme. The probe used was 4-(2-iodoacetamido)-2,2,6,6 tetramethylpiperidine-1-oxyl (IASL), which reacts specifically with sulfhydryl 1 of the myosin head. In the absence of nucleotide, the probe remains strongly immobilized (rigidly attached to the myosin head) so that no nanosecond rotational motions are detectable. When MgADP is added to IASL-labeled myosin filaments (T = 20 degrees C), the probe mobility increases slightly. During steady-state MgADP hydrolysis (T = 20 degrees C), the probe undergoes large amplitude nanosecond rotational motion. These results are consistent with previous studies of myosin monomers, heavy meromyosin, and myosin subfragment 1. Isoclinic points observed in overlays of sequential EPR spectra recorded during ATP hydrolysis strongly suggest that the probes fall into two motional classes, separated by approximately an order of magnitude in effective rotational correlation time. Both of the observed states are distinct from the conformation of myosin in the absence of nucleotides, and the spectrum of the less mobile population is indistinguishable from that observed in the presence of MgADP. The addition of ADP and vanadate to IASL-myosin gives rise to two motional classes virtually identical with those observed in the presence of ATP, but the relative concentrations of the spin populations are significantly different. We have quantitated the percentage of myosin in each motional state during ATP hydrolysis. The result agrees well with the predicted percentages in the two predominant chemical states in the myosin ATPase cycle. Spectra obtained in the presence of nucleotide analogues permit us to assign the conformational states to specific chemical states. We propose that the two motional classes represent two distinct local conformations of myosin that are in exchange with one another during the ATP hydrolysis reaction cycle. PMID- 3030404 TI - Spectrochemical studies on the blue copper protein azurin from Alcaligenes denitrificans. AB - Spectroscopic and electrochemical studies, incorporating electronic spectra, electron paramagnetic resonance (EPR) spectra, resonance Raman (RR) spectra, and measurements of the redox potential, have been carried out on the blue copper protein azurin, from Alcaligenes denitrificans. These data are correlated with the refined crystal structure of this azurin and with corresponding data for other blue copper proteins. The electronic spectrum, characterized by an intense (epsilon = 5100 M-1 cm-1) charge-transfer band at 619 nm, the EPR spectral parameters (g perpendicular = 2.059, g parallel of = 2.255, A parallel of = 60 X 10(-4) cm-1), and the resonance Raman spectrum are similar to those obtained from other azurins and from plastocyanins. Both the electronic spectrum and the EPR spectrum are unchanged over the pH range 4-10.5, but major changes occur above pH 12 and below pH 3.5. A small reversible change occurs at pH approximately 11.4. In the RR spectrum the Cu-S stretching mode is shown to contribute to all of the five principal RR peaks. Deuterium substitution produces shifts in at least seven of the peaks; these shifts may be attributable, at least in part, to the NH...S hydrogen bond to the copper-ligated Cys-112. Measurements of the redox potential, using spectroelectrochemical methods, over the temperature range 4.8-40.0 degrees C, give values for delta H0' and delta S0' of -55.6 kJ mol-1 and -97.0 J K-1 mol 1, respectively. The redox potential of A. denitrificans azurin at pH 7.0, Eo', is 276 mV. These data are interpreted in terms of a copper site, in azurin, comprising three strong bonds, in an approximately trigonal plane, from Cys-112, His-46, and His-117 and much longer axial approaches from Met-121 and the peptide carbonyl oxygen of Gly-45. Spectral differences within the azurin family and between azurin and plastocyanin are attributed to differences in the strengths of these axial interactions. Likewise, the distinctly lower Eo values for azurins, as compared with plastocyanins, are related to the more copper(II)-like site in azurin [with a weaker Cu-S(Met) interaction and a Cu-O interaction not found in plastocyanin]. On the other hand, the relative constancy of the EPR parameters between azurin and plastocyanin suggests they are not strongly influenced by weakly interacting axial groups. PMID- 3030405 TI - Predicted structures of cAMP binding domains of type I and II regulatory subunits of cAMP-dependent protein kinase. AB - The mammalian cAMP-dependent protein kinases have regulatory (R) subunits that show substantial homology in amino acid sequence with the catabolite gene activator protein (CAP), a cAMP-dependent gene regulatory protein from Escherichia coli. Each R subunit has two in-tandem cAMP binding domains, and the structure of each of these domains has been modeled by analogy with the crystal structure of CAP. Both the type I and II regulatory subunits have been considered, so that four cAMP binding domains have been modeled. The binding of cAMP in general is analogous in all the structures and has been correlated with previous results based on photolabeling and binding of cAMP analogues. The model predicts that the first cAMP binding domain correlates with the previously defined fast dissociation site, which preferentially binds N6-substituted analogues of cAMP. The second domain corresponds to the slow dissociation site, which has a preference for C8-substituted analogues. The model also is consistent with cAMP binding in the syn conformation in both sites. Finally, this model has targeted specific regions that are likely to be involved in interdomain contacts. This includes contacts between the two cAMP binding domains as well as contacts with the amino-terminal region of the R subunit and with the catalytic subunit. PMID- 3030406 TI - Yeast cytochrome c peroxidase: mutagenesis and expression in Escherichia coli show tryptophan-51 is not the radical site in compound I. AB - Using oligonucleotide-directed site-specific mutagenesis, we have constructed a system for the mutation and expression of yeast cytochrome c peroxidase (CCP, EC 1.11.1.5) in Escherichia coli and applied it to test the hypothesis that Trp-51 is the locus of the free radical observed in compound I of CCP [Poulos, T. L., & Kraut, J. (1980) J. Biol. Chem. 255, 8199-8205]. The system was created by substituting a CCP gene modified by site-directed mutagenesis, CCP(MI), for the fol gene in a vector previously used for mutagenesis and overexpression of dihydrofolate reductase. E. coli transformed with the resulting plasmid produced the CCP(MI) enzyme in large quantities, more than 15 mg/L of cell culture, of which 10% is holo- and 90% is apo-CCP(MI). The apoenzyme was easily converted to holoenzyme by the addition of bovine hemin. Purified CCP(MI) has the same catalytic activity and spectra as bakers' yeast CCP. A mutation has been made in CCP(MI), Trp-51 to Phe. The Phe-51 mutant protein CCP(MI,F51) is fully active, and the electron paramagnetic resonance (EPR) spectrum, at 89 K, of its oxidized intermediate, compound I, displays a strong sharp resonance at g = 2.004, which is very similar to the signal observed for compound I of both bakers' yeast CCP and CCP(MI). However, UV-visible and EPR spectroscopy revealed that the half-life of CCP(MI,F51) compound I at 23 degrees C is only 1.4% of that observed for the compound I forms of CCP(MI) or bakers' yeast CCP. Thus, Trp-51 is not necessary for the formation of the free radical observed in compound I but appears to exert a significant influence on its stability. PMID- 3030407 TI - DNA structure equilibria in the human c-myc gene. AB - We have employed analytical S1 nuclease analysis to identify sites with altered DNA secondary structure in the human c-myc gene. We have mapped several sites of that kind in vitro at one-base resolution but have focused our attention on one particularly stable conformational isomer which occurs approximately 270 base pairs upstream from the preferred transcription origin. We have analyzed the kinetics of that conformational equilibrium as a function of supercoil density and enzyme concentration and find that DNA structure in this region is adequately modeled as a two-state equilibrium between an undistorted (S1 nuclease insensitive) and a distorted (S1-sensitive) state. We find that at fixed supercoil density, S1 nuclease cleavage at this DNA segment can be altered in vitro by a DNA sequence change as far away as 1500 bases. We also find that the S1 nuclease cleavage at this site can be dramatically enhanced by the binding of small RNA molecules. On the basis of an analysis of S1 cutting kinetics and an analysis of DNA sequence at the S1 cleavage site, we conclude that RNA may bind directly to DNA, thereby shifting the underlying conformational equilibrium. Together, these data suggest that as a class, short RNA molecules could serve as site-specific regulatory elements in the myc gene and elsewhere. PMID- 3030409 TI - Purification and structural characterization of herpes simplex virus glycoprotein C. AB - Purification of herpes simplex virus glycoprotein C (gC) in microgram amounts yielded sufficient material for an analysis of its secondary structure. Purification was facilitated by using the mutant virus gC-3, which bears a point mutation that interrupts the putative hydrophobic membrane anchor sequence, causing the secretion of gC-3 protein into the cell culture medium. gC-3 protein was purified by size fractionation of concentrated culture medium from infected cells on a gel filtration column of Sephacryl S-200, followed by immunoaffinity chromatography on a column constructed of gC-specific monoclonal antibodies cross linked to a protein A-Sepharose CL-4B matrix. Purified gC-3 had a molecular weight of 130,000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the size expected for gC, was reactive with gC-specific monoclonal antibodies in protein immunoblots, and contained amino acid sequences characteristic of gC as determined by radiochemical amino acid microsequence analyses. Polyclonal antisera obtained from a rabbit immunized with gC-3 reacted with wild-type gC in immunoprecipitation, enzyme immunoassay, and immunoelectroblot (western blot) assays. Deglycosylation by treatment with trifluoromethanesulfonic acid reduced the molecular weight of gC-3 by approximately 35%. Analyses of both native and deglycosylated gC-3 by Raman spectroscopy showed that the native molecule consists of about 17% alpha-helix, 24% beta-sheet, and 60% disordered secondary structures, whereas deglycosylated gC-3 consists of about 8% alpha-helix, 10% beta-sheet, and 81% disordered structures. These data were in good agreement with the 11% alpha-helix, 18% beta sheet, 61% beta-turn, and 9% disordered structures calculated from Chou-Fasman analysis of the primary sequence of gC-3. PMID- 3030408 TI - Respective role of each of the purine N7 nitrogens of 5'-O-triphosphoadenylyl(2'- --5')adenylyl(2'----5')adenosine in binding to and activation of the RNase L of mouse cells. AB - Through a combination of chemical and enzymatic approaches a series of sequence specific tubercidin-substituted ppp5'A2'p(5'A2'p)n5'A (n = 1 to about 10; 2-5A) analogues were generated. In addition to the previously developed methodology of Imai and Torrence [Imai, J., & Torrence, P.F. (1985) J. Org. Chem. 50, 1418 1420], a new approach to synthesis of 2',5'-linked oligonucleotides utilized adenosine in 3',5' linkage as a precursor to the targeted 5'-terminus of the desired product. For instance, A3'p5'A could be condensed under conditions of lead ion catalysis with tubercidin 5'-phosphate to give A3'p5'A2'p5'(c7A). Treatment with the 3',5'-specific nuclease P1 led to p5'A2'p5'(c7A). The combined use of the above procedures led to the synthesis of p5'(c7A)2'p5'A2'p5'A, p5'A2'p5'(c7A)2'p5'A, p5'A2'p5'A2'p5'(c7A), and p5'A2p5'(c7A)2'p5'(c7A), which were converted to their corresponding 5'-triphosphates by the usual methods. Evaluation of these analogues for their ability to bind to and activate the 2-5A dependent endonuclease (RNase L) of mouse L cells showed that there were small changes (less than or equal to 10-fold) in the ability of the four tubercidin analogues to bind to RNase L. However, whenever the first and/or third adenosine nucleotide units were replaced by tubercidin, a dramatic decrease in ability to activate RNase L occurred. Only the second (from the 5'-terminus) adenosine residue could be replaced by tubercidin without any effect on RNase L activation ability. PMID- 3030410 TI - Multiple equilibria binding treatment of lipid and detergent interactions with membrane proteins. Application to cytochrome c oxidase solubilized in cholate. AB - A modified multiple binding equilibria treatment is presented that allows determination of thermodynamic parameters of the interaction of phospholipids with integral membrane proteins solubilized in excess detergent. Lipid binding is modeled as a series of exchange reactions between lipid molecules and detergent molecules at the hydrophobic protein surface. A general equation is derived which expresses a relative association constant (K) and the total number of contact sites at the lipid-protein interface (N) in terms of experimentally measurable variables. A useful simplification of the general equation occurs when the amount of detergent is high relative to the total number of lipid binding sites in the sample. Computer simulations show that in cases we have examined there appears to be an experimentally accessible range of detergent to protein molar ratios where the approximation at high detergent is useful for analyzing experimental data. This model is used to examine the competition between cholate and spin-labeled phospholipids for the hydrophobic surfaces of bovine heart cytochrome c oxidase. We find, for example, that K = 12 +/- 2 for phosphatidylcholine relative to cholate (i.e., the cholate molecules are relatively easily displaced by membrane lipids). This helps to explain the experimental observation that cholate is an effective detergent both for solubilizing cytochrome c oxidase and for reconstituting this protein into a defined lipid bilayer environment. An excess of cholate readily displaces almost all of the native phospholipids, and the protein is dispersed in cholate micelles. However, when phospholipids are added back, the cholate molecules at the protein surface are replaced because of the higher relative binding of the phospholipids. Observed differences between the behavior of phosphatidylcholine and phosphatidylglycerol suggest that reconstitution in cholate is a selective process in which detergent molecules in localized areas on the protein surface are more readily displaced by certain phospholipids. PMID- 3030411 TI - 13C and 31P NMR study of gluconeogenesis: utilization of 13C-labeled substrates by perfused liver from streptozotocin-diabetic and untreated rats. AB - The metabolism of 13C-labeled substrates was followed by 13C and 31P NMR in perfused liver from the streptozotocin-treated rat model of insulin-dependent diabetes. Comparison was made with perfused liver from untreated littermates, fasted either 24 or 12 h. The major routes of pyruvate metabolism were followed by a 13C NMR approach that provided for the determination of the metabolic fate of several substances simultaneously. The rate of gluconeogenesis was 2-4-fold greater and beta-hydroxybutyrate production was 50% greater in liver from the chronically diabetic rats as compared with the control groups. Large differences in the distribution of 13C label in hepatic alanine were measured between diabetic and control groups. The biosyntheses of 13C-labeled glutathione and N carbamoylaspartate were monitored in time-resolved 13C NMR spectra of perfused liver. Assignments for the resonances of glutathione and N-carbamoylaspartate were made with the aid of 13C NMR studies of perchloric acid extracts of the freeze-clamped livers. 13C NMR spectroscopy of the perfusates provided a convenient, rapid assay of the rate of oxidation of [2-13C]ethanol, the hepatic output of [2-13C]acetaldehyde, and the accumulation of [2-13C]acetate in the perfusate. By 31P NMR spectroscopy, carbamoyl phosphate was measured in all diabetic livers and an unusual P,P'-diesterified pyrophosphate was observed in one-fourth of the diabetic livers examined. Neither of these phosphorylated metabolites was detected in control liver. Both 13C and 31P NMR were useful in defining changes in hepatic metabolism in experimental diabetes. PMID- 3030412 TI - Effects of insulin on perfused liver from streptozotocin-diabetic and untreated rats: 13C NMR assay of pyruvate kinase flux. AB - The effects of insulin in vitro on perfused liver from streptozotocin-diabetic rats and their untreated littermates during gluconeogenesis from either [3 13C]alanine + ethanol or [2-13C]pyruvate + NH4Cl + ethanol were studied by 13C NMR. A 13C NMR determination of the rate of pyruvate kinase flux under steady state conditions of active gluconeogenesis was developed; this assay includes a check on the reuse of recycled pyruvate. The preparations studied provided gradations of pyruvate kinase flux within the confines of the assay's requirement of active gluconeogenesis. By this determination, the rate of pyruvate kinase flux was 0.74 +/- 0.04 of the gluconeogenic rate in liver from 24-h-fasted controls; in liver from 12-h-fasted controls, relative pyruvate kinase flux increased to 1.0 +/- 0.2. In diabetic liver, this flux was undetectable by our NMR method. Insulin's hepatic influence in vitro was greatest in the streptozotocin model of type 1 diabetes: upon treatment of diabetic liver with 7 nM insulin in vitro, a partial reversal of many of the differences noted between diabetic and control liver was demonstrated by 13C NMR. A major effect of insulin in vitro upon diabetic liver was the induction of a large increase in the rate of pyruvate kinase flux, bringing relative and absolute fluxes up to the levels measured in 24-h-fasted controls. By way of comparison, the effects of ischemia on diabetic liver were studied by 13C NMR to test whether changes in allosteric effectors under these conditions could also increase pyruvate kinase flux. A large increase in this activity was demonstrated in ischemic diabetic liver. PMID- 3030414 TI - Dual pathways in muscarinic receptor stimulation of phosphoinositide hydrolysis. AB - The relationships between phosphoinositide hydrolysis induced by various muscarinic agonists and by membrane depolarization agents were investigated in rat cerebral cortex and heart atrium slices. In both preparations, phosphoinositide hydrolysis was stimulated by a combination of carbamylcholine and membrane depolarization with 40 mM K+ in a synergistic fashion. The synergism was more pronounced at lower external calcium ion concentrations and was sensitive to verapamil. Lower external calcium ion concentrations were required for demonstration of the synergism in heart atrium slices than in cerebral cortex slices. The carbamylcholine-induced stimulation was only partially additive with membrane depolarization via Na+ channel gating by batrachotoxin. In addition, K+ depolarization eliminated the sensitivity of carbamylcholine-stimulated phosphoinositide hydrolysis to the sodium channel blocker tetrodotoxin. Our results suggest that muscarinically stimulated phosphoinositide hydrolysis in rat cerebral cortex and heart atrium slices may occur by dual pathways which interact synergistically and that only one of the pathways is depolarization-dependent. Different muscarinic agonists could preferentially utilize these pathways, thus perhaps explaining their different potencies in stimulating phosphoinositide hydrolysis. PMID- 3030413 TI - Identification of purine deoxyribonucleoside kinases from human leukemia cells: substrate activation by purine and pyrimidine deoxyribonucleosides. AB - Cell extracts from human leukemic T lymphoblasts and myeloblasts were chromatographed on DEAE-cellulose columns to separate purine deoxyribonucleoside, deoxyadenosine (dAdo) and deoxyguanosine (dGuo), phosphorylating activities. Three distinct purine deoxyribonucleoside kinases, a deoxycytidine (dCyd) kinase, an adenosine (Ado) kinase, and a deoxyguanosine (dGuo) kinase (the latter appears to be localized in mitochondria), were resolved. dCyd kinase contained the major phosphorylating activity for dAdo, dGuo, and 9-beta-D-arabinofuranosyladenine (ara-A). Ado kinase represented a second kinase for dAdo and ara-A while a third kinase for dAdo was found in mitochondria. dCyd kinase was purified about 2000 fold with ion-exchange, affinity, and hydrophobic chromatographies. On gel electrophoresis, both dCyd and dAdo phosphorylating activities comigrated, indicating that the activities are associated with the same protein. The enzyme showed a broad pH optimum ranging from pH 6.5 to pH 9.5. Divalent cations Mg2+, Mn2+, and Ca2+ stimulated dCyd kinase activity; Mg2+ produced the maximal activity. dCyd kinase from either lymphoid or myeloid cells showed broad substrate specificity. The enzyme used several nucleoside triphosphates, but ATP, GTP, and dTTP were the best phosphate donors. dCyd was the best nucleoside substrate, since dCyd kinase had an apparent Km of 0.3, 85, 90, and 1400 microM for dCyd, dAdo, dGuo, and ara-A, respectively. The enzyme exhibited substrate activation with both pyrimidine and purine deoxyribonucleosides, suggesting that there is more than one substrate binding site on the kinase. These studies show that, in lymphoblasts and myeloblasts, purine deoxyribonucleosides and their analogues are phosphorylated by dCyd kinase, Ado kinase, and dGuo kinase. PMID- 3030415 TI - Respiratory proteins from extremely thermophilic, aerobic bacteria. PMID- 3030416 TI - Intermediate steps in the reaction of cytochrome oxidase with molecular oxygen. PMID- 3030417 TI - Membrane catalysis: synchronous multielectron reactions at the interface between two liquid phases. Bioenergetic mechanisms. AB - Kinetics of multi-electron reactions at the interface between two immiscible liquids are considered. Calculations of the energy of solvent reorganization, of the work required to bring reactants and reaction products together, and of the electrostatic contributions to the Gibbs free energy of the reaction during electron transfer between reactants which are in different dielectric media are reported. Conditions under which the free energy of activation of the interfacial reaction of electron transfer decreases are established. The influence of the distance between reactants and of the dielectric permittivity of the non-aqueous phase on the solvent reorganization energy value is studied. Conditions under which multielectron reactions at the interface proceed are discussed. The biophysics and biochemistry of photosynthesis and respiration are considered as examples of multielectron processes. PMID- 3030418 TI - Substrate-dependent functional defects and altered mitochondrial respiratory capacity in hearts from guinea pigs with iron deficiency anemia. AB - Iron deficiency anemia was induced by dietary means in weanling guinea pigs. A 25% higher ventricular wall mass per 100 g body mass was seen after 6 weeks of feeding. Myocardial performance was determined in isolated perfused hearts using an isovolumic Langendorff preparation. All hearts exhibited a 25% decrease in left ventricular developed pressure (LVDP) and decreased dP/dt when substrate was switched from 10 mM pyruvate to 16.6 mM glucose. The glucose reduction in LVDP resulted from decreased systolic pressure, which completely reversed when hearts again metabolized pyruvate. With glucose as substrate, left ventricular developed pressure-end diastolic volume relationships were indistinguishable. However, with pyruvate, iron-deficient hearts appeared to be less responsive to the increased energy demands required by elevated diastolic volumes. Rates of state 3 respiration were 18% below control with glutamate + malate as substrate, and 38% lower with pyruvate + malate in mitochondria isolated from anemic animals. No differences in respiration were noted with succinate. Cytochrome a + a3 content, cytochrome oxidase activity and total mitochondrial protein content appeared to be unchanged. In contrast, cytochromes b, c + c1, and the flavoproteins were significantly decreased. The data suggest that iron deficiency anemia induces cardiac hypertrophy with a fixed but defective mitochondrial population, potentially placing the heart in an energetic imbalance. These differences in mitochondrial function were expressed by decreased myocardial performance when the heart metabolizes pyruvate, an exclusively aerobic substrate. PMID- 3030419 TI - Specific inhibition of ATP-ADP translocase in cardiac mitoplasts by antibodies against mitochondrial creatine kinase. AB - Mitochondrial creatine kinase was purified from rat hearts and used to produce antibodies in chicken and rabbits. Antibodies were purified to a high degree of homogeneity by an affinity chromatography method. Chicken antibodies against mitochondrial creatine kinase inhibited this enzyme in rat-heart mitochondrial inner membrane and matrix preparation, and simultaneously blocked oxidative phosphorylation. Under these conditions respiratory chain activities remained unchanged, but adenine nucleotide translocase was inhibited. Removal of mitochondrial creatine kinase from the membrane by pretreatment with 0.15 M KCl and 20 mM ADP completely abolished the effect of antibodies against mitochondrial creatine kinase on oxidative phosphorylation. Noninhibitory antibodies from rabbit with high affinity to rat mitochondrial creatine kinase inhibited neither creatine kinase activity nor oxidative phosphorylation. These data show close and specific spatial arrangement of mitochondrial creatine kinase and adenine nucleotide translocase in mitochondria. It is supposed that there is a fixed orientation of these proteins in the cardiolipin domain in the membrane and that their interaction may occur by a frequent collision due to their lateral movement. PMID- 3030420 TI - Cytochrome a3 deficiency in human achondroplasia. AB - Mitochondria prepared from tissue culture cells (skin fibroblasts) from normal subjects and subjects with homozygous achondroplasia were studied to determine the concentrations of cytochromes a and a3 in the preparations. Cytochrome a3 was markedly decreased (80%) in the achondroplastic preparations with cytochrome a present in normal amounts. Determination of total heme a (as the pyridine hemochromogen) in the normal and achondroplastic preparations demonstrated that the observed decrease in concentration of cytochrome a3 in the achondroplastic preparations was due to an absence of cytochrome a3 and not to a change in its absorbancy (extinction coefficient). The decreased concentrations of cytochrome a3 in the achondroplastic cells may decrease the reactivity or affinity of the mitochondrial oxidative systems for oxygen and result in the phenotypic expression of the disease. PMID- 3030421 TI - Separation of luminal and abluminal membrane enriched domains from cultured bovine aortic endothelial cells: monoclonal antibodies specific for endothelial cell plasma membranes. AB - Two kinds of membrane (luminal and abluminal membrane domains) fractions have been isolated from bovine aortic endothelial cells by fractionation of whole cell homogenate on discontinuous sucrose density gradients. The luminal membrane domain was enriched 12-16-fold for angiotensin-converting enzyme activity and 8 10-fold in alkaline phosphatase activity. The abluminal membrane domain displayed an enrichment of 8-fold in (Na+ + K+)-ATPase activity. Both of the membrane domains were minimally contaminated with mitochondria, microsomes and Golgi bodies, as assessed by their corresponding marker enzyme activities. 125I labeling of endothelial cell monolayers by the Enzymo-Bead lactoperoxidase catalyzed iodination procedure, followed by isolation of membranes, revealed that the radioactivity was predominantly associated with membranes enriched in angiotensin-converting enzyme activity, corresponding to the luminal membrane domain. However, when cells were radioiodinated in suspension culture, radioactivity was found equally associated in both the luminal and abluminal membrane fractions. Electron microscopy of freeze-fractured and sectioned material showed both luminal and abluminal membrane domains to be in the form of vesicles varying in size from 100 to 400 nm in diameter. To characterize the separation of endothelial cell membrane domains, we have attempted to prepare monoclonal antibodies specific for endothelial cells. Several clones were obtained, producing antibodies which bound to endothelial cells of arterial, venous and capillary origin. Two antibodies of these clones, XIVC6 and XVD2, were studied in more detail. In the ELISA assay, these antibodies reacted with bovine vascular endothelial cells, but not with human umbilical cord endothelial cells, nor with bovine corneal endothelial cells, smooth muscle cells or fibroblasts. Both of these antibodies are directed against an antigen of approximately 130 kDa, under reducing and non-reducing conditions, as assayed by the immunoprecipitation method. Western blot analysis of luminal and abluminal membrane fractions revealed that only MAb XVD2 reacted with an antigen, indicating that the antibody XIVC6 is directed against an epitope which is denatured by SDS. Moreover, MAb XVD2 preferentially reacted with the luminal membrane compared to the abluminal membrane domain of the endothelial cell. These monoclonal antibodies do not react with platelet membrane proteins, indicating that this 130 kDa membrane antigen is not common to both endothelial cells and platelets.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3030422 TI - Binding and activation of rod outer segment phosphodiesterase and guanosine triphosphate binding protein by disc membranes: influence of reassociation method and divalent cations. AB - Attempts to optimize the recovery of light-stimulated phosphodiesterase activity following reassociation of the hypotonically extractable proteins derived from retinal rod segments with hypotonically stripped disc membranes lead to the following observations: the best reassociations were obtained by mixing proteins and stripped disc membranes under hypotonic conditions and slowly increasing the salt concentration; the binding of G-protein and phosphodiesterase to stripped disc membrane occurs in less than 5 minutes and the recovery of light-stimulated phosphodiesterase activation in response to subsaturating stimulus levels requires 2-3 h to plateau. Stripped disc membranes and proteins were reassociated in 'isotonic' buffers containing KCl/NaCl, KCl/NaCl plus Mg2+, or KCl/NaCl plus Ca2+. Large fractional rhodopsin bleaches produced nearly identical light stimulated phosphodiesterase activities in each of these samples and in the control rod outer segment membranes. Rod outer segment membranes and reassociated stripped disc membrane samples containing divalent cations showed similar phosphodiesterase activities in response to low fractional rhodopsin bleaches (e.g. less than or equal to 0.1%), however, samples devoid of divalent cations during reassociation required rhodopsin bleaches up to 10-fold larger to elicit comparable phosphodiesterase activities. These results suggest that not all phosphodiesterase and/or G-protein molecules bound to the disc membrane surface are equivalent with regard to their efficiency of activation by bleached rhodopsin and that divalent cations can modulate the distribution of G-protein and/or phosphodiesterase between these populations. PMID- 3030423 TI - Lipid-dependent differential effects of stereoisomers of anesthetic alcohols. AB - The cis- and trans-alkenols are equally potent general anesthetics but, respectively, lower and raise the gel-to-liquid crystalline phase transition temperature of saturated phosphatidylcholines (Pringle, M.J. and Miller, K.W. (1978) Biochem. Biophys. Res. Commun. 85, 1191-1198). Here we show that although this differential effect is somewhat reduced when a double bond is introduced into the sn-2 position of phosphatidylcholine, it is abolished when the ethanolamine head group is substituted for the choline head group in dimyristoyl lipids at neutral pH. At high pH, however, dimyristoylphosphatidylethanolamine assumes a negative charge, and its phase transition temperature drops to a value close to that for the corresponding phosphatidylcholine. Under these conditions the differential effect of the alkenol isomers is restored; the cis-alkenol lowers, while the trans-alkenol raises, the phase transition temperature of deprotonated dimyristoylphosphatidylethanolamine. Thus, the differential effects of cis- and trans-alkenols on the gel-to-liquid crystalline phase transition are dependent on the physical chemical characteristics of the polar region of the perturbed lipid species, but only weakly on that of the acyl region. PMID- 3030424 TI - The interaction of dietary fatty acid and cholesterol on catecholamine-stimulated adenylate cyclase activity in the rat heart. AB - Diets supplemented with high levels of saturated or unsaturated fatty acids supplied by addition of sheep kidney fat or sunflower seed oil, respectively, were fed to rats with or without dietary cholesterol. The effects of these diets on cardiac membrane lipid composition, catecholamine-stimulated adenylate cyclase and beta-adrenergic receptor activity associated with cardiac membranes, were determined. The fatty acid-supplemented diets, either with or without cholesterol, resulted in alterations in the proportion of the (n-6) to (n-3) series of unsaturated fatty acids, with the sunflower seed oil increasing and the sheep kidney fat decreasing this ratio, but did not by themselves significantly alter the ratio of saturated to unsaturated fatty acids. However, cholesterol supplementation resulted in a decrease in the proportion of saturated and polyunsaturated fatty acids and a dramatic increase in oleic acid in cardiac membrane phospholipids irrespective of the nature of the dietary fatty acid supplement. The cholesterol/phospholipid ratio of cardiac membrane lipids was also markedly increased with dietary cholesterol supplementation. Although relatively unaffected by the nature of the dietary fatty acid supplement, catecholamine-stimulated adenylate cyclase activity was significantly increased with dietary cholesterol supplementation and was positively correlated with the value of the membrane cholesterol/phospholipid ratio. Although the dissociation constant for the beta-adrenergic receptor, determined by [125I](-) iodocyanopindolol binding, was unaffected by the nature of the dietary lipid supplement, the number of beta-adrenergic receptors was dramatically reduced by dietary cholesterol and negatively correlated with the value of the membrane cholesterol/phospholipid ratio. These results indicate that the activity of the membrane-associated beta-adrenergic/adenylate cyclase system of the heart can be influenced by dietary lipids particularly those altering the membrane cholesterol/phospholipid ratio and presumably membrane physico-chemical properties. In the face of these dietary-induced changes, a degree of homeostasis was apparent both with regard to membrane fatty acid composition in response to an altered membrane cholesterol/phospholipid ratio, and to down regulation of the beta-adrenergic receptor in response to enhanced catecholamine-stimulated adenylate cyclase activity. PMID- 3030425 TI - Estimation of spin probe clustering in biological membranes. AB - An iterative spectral subtraction technique has been developed which accurately estimates the proportion of 'dilute' and 'clustered' I(12, 3) (i.e., 5-nitroxide stearate) in human erythrocyte ghosts at 37 degrees C, even if subtractant spectra free from probe-probe interactions cannot be measured due to technical limitations. Gordon et al. ((1985) J. Membrane Biol. 84, 81-95) earlier showed that I(12, 3) occupies a class of high-affinity sites in ghosts at probe/total lipid ratios (P/L) less than 1/2250. Saturation occurs with increasing probe concentration, and, at higher loading, the probe inserts itself at initially dilute sites to form membrane-bound clusters of variable size. Although this model allows determination of the dilute/clustered probe ratio, it requires subtraction of experimental spectra with a 'magnetically dilute' spectrum obtained using P/L less than 1/4600. The new methodology accurately profiles the % probe clustering in human erythrocyte ghosts over the entire P/L range, even if the lowest P/L for the subtractant spectrum contains substantial probe-probe interactions (i.e., P/L of 1/604 or 1/303). Application of either the subtraction technique in Gordon et al. (1985) or the iterative subtraction protocol described here should allow determination of probe clustering in a wide range of I(12, 3) labeled biological membranes. PMID- 3030427 TI - The effect of D-penicillamine on myeloperoxidase: formation of compound III and inhibition of the chlorinating activity. AB - The inhibitory effect of the anti-arthritic drug D-penicillamine on the formation of hypochlorite (HOCl) by myeloperoxidase from H2O2 and Cl- was investigated. When D-penicillamine was added to myeloperoxidase under turnover conditions, Compound III was formed, the superoxide derivative of the enzyme. Compound III was not formed when D-penicillamine was added in the presence of EDTA or in the absence of oxygen. However, when H2O2 was added to myeloperoxidase, D penicillamine and EDTA, Compound III was formed. Therefore it is concluded that formation of Compound III is initiated by metal-catalysed oxidation of the thiol group of this anti-arthritic drug, resulting in formation of superoxide anions. Once Compound III is formed, a chain reaction is started via which the thiol groups of other D-penicillamine molecules are oxidized to disulphides. Concomitantly, Compound I of myeloperoxidase would be reduced to Compound II and superoxide anions would be generated from oxygen. This conclusion is supported by experiments which showed that formation of Compound III of myeloperoxidase by D penicillamine depended on the chloride concentration. Thus, an enzyme intermediate which is active in chlorination (i.e. Compound I) participated in the generation of superoxide anions from the anti-arthritic drug. From the results described in this paper it is proposed that D-penicillamine may exert its therapeutic effect in the treatment of rheumatoid arthritis by scavenging HOCl and by converting myeloperoxidase to Compound III, which is inactive in the formation of HOCl. PMID- 3030428 TI - Photoaffinity labelling of the 2-oxoglutarate binding site of prolyl 4 hydroxylase with 5-azidopyridine-2-carboxylic acid. AB - The synthesis of the photoaffinity label 5-azidopyridine-2-carboxylic acid is described. The 2-oxoglutarate analogue photoaffinity label is a competitive inhibitor with respect to 2-oxoglutarate with a Ki value of 9 X 10(-3) M. Upon ultraviolet irradiation, 5-azidopyridine-2-carboxylic acid inactivated prolyl 4 hydroxylase irreversibly by up to 50%. The extent of inactivation depended on the 5-azidopyridine-2-carboxylic acid concentration and the irradiation time. Inactivation was prevented in the presence of an excess of 2-oxoglutarate. It is concluded that the 5-azidopyridine-2-carboxylic acid became covalently bound to the alpha subunit of prolyl 4-hydroxylase, as the alpha subunit of the photoaffinity labelled enzyme had a decreased electrophoretic mobility in polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate. PMID- 3030426 TI - Proton/hydroxide conductance through phospholipid bilayer membranes: effects of phytanic acid. AB - Mechanisms of proton/hydroxide conductance (GH/OH) were investigated in planar (Mueller-Rudin) bilayer membranes made from decane solutions of phospholipids or phospholipids plus phytanic acid (a 20-carbon, branched chain fatty acid). At neutral pH, membranes made from diphytanoylphosphatidylcholine or bacterial phosphatidylethanolamine had GH/OH values in the range of (2-5) X 10(-9) S X cm 2, corresponding to H+/OH- 'net' permeabilities of about (0.4-1.0) X 10(-5) cm X s-1. GH/OH was inhibited by serum albumin, phloretin, glycerol and low pH, but was increased by chlorodecane and voltage greater than 80 mV. Water permeability and GH/OH were not correlated, suggesting that water and H+/OH- cross the membrane by separate pathways. Addition of phytanic acid to the phospholipids caused an increase in GH/OH which was proportional to the first power of the phytanic acid concentration. In membranes containing phytanic acid, GH/OH was inhibited by albumin, phloretin, glycerol and low pH, but was increased by chlorodecane and voltages greater than 80 mV. The results suggest that phytanic acid acts as a simple (A- type) proton carrier. The qualitative similarities between the behavior of GH/OH in unmodified and phytanic-acid containing membranes suggest that phospholipids may contain weakly acidic contaminants which cause most of GH/OH at pH greater than 4. However, there is also a significant background (pH independent) GH/OH which may be due to hydrogen-bonded water chains. The ability of phytanic acid to act as a proton carrier may help to explain the toxicity of phytanic acid in Refsum's disease, a metabolic disorder in which phytanic acid accumulates to high levels in plasma, cells and tissues. PMID- 3030429 TI - The inhibition of cytochrome oxidase by diaminomaleonitrile. AB - Diaminomaleonitrile, a tetramer of cyanide, was examined as a possible antagonist to cyanide inhibition of cytochrome oxidase (EC 1.9.3.1). This compound was found to inhibit cytochrome oxidase in vitro; however, despite their structural similarities, diaminomaleonitrile and cyanide inhibit cytochrome oxidase by different mechanisms and bind to the enzyme at different sites. Diaminomaleonitrile inhibition of cytochrome oxidase is described in terms of a partially competitive mechanism. Biological oxidation of diaminomaleonitrile may lead to the formation of cyanide. PMID- 3030430 TI - Papain fragmentation of the gastric (H+ + K+)-ATPase. AB - Membrane-bound (H+ + K+)-ATPase purified from hog gastric mucosa was exposed to limited papain digestion. Such treatment resulted in a rapid inhibition of the K+ stimulated adenosine triphosphatase and p-nitrophenyl phosphatase activities, with about 90% of these activities lost after 3 min incubation at 37 degrees C with 0.1 units of papain per mg of enzyme protein. Parallel to the inhibition of the enzyme activities, there was a production of a 77 kDa membrane-bound fragment containing the aspartyl phosphate residue of the phospho-intermediate. This fragment accounted for about 45% of the total enzyme protein after the 3 min papain treatment. The digestion barely affected the steady-state level of phosphorylation, allowed the aspartyl phosphate of the 77 kDa fragment to undergo the transition to the E2P form, and did not significantly alter the fraction of ADP-sensitive phosphoenzyme. The presence of KCl, however, depressed the steady state level of phosphoenzyme formed from [gamma-32P]ATP considerably less than that of the control enzyme. With further exposure to papain the 77 kDa peptide became fragmented into a 28 kDa soluble peptide that retained the phosphorylating site. Binding of fluorescein 5'-isothiocyanate (FITC) to the native enzyme did not affect the sites of papain hydrolysis because the same peptide fragments were obtained. The FITC reaction site was also in the 28 kDa soluble peptide fragment. PMID- 3030431 TI - Electron paramagnetic resonance and spectrophotometric studies of the peroxide compounds of manganese-substituted horseradish peroxidase, cytochrome-c peroxidase and manganese-porphyrin model complexes. AB - Peroxide compounds of manganese protoporphyrin IX and its complexes with apo horseradish peroxidase and apocytochrome-c peroxidase were characterized by electronic absorption and electron paramagnetic resonance spectroscopies. An intermediate formed upon titration of Mn(III)-horseradish peroxidase with hydrogen peroxide exhibited a new electron paramagnetic resonance absorption at g = 5.23 with a definite six-lined 55Mn hyperfine (AMn = 8.2 mT). Neither a porphyrin pi-cation radical nor any other radical in the apoprotein moiety could be observed. The reduced form of Mn-horseradish peroxidase, Mn(II)-horseradish peroxidase, reacted with a stoichiometric amount of hydrogen peroxide to form a peroxide compound whose electronic absorption spectrum was identical with that formed from Mn(III)-horseradish peroxidase. The electronic state of the peroxide compound of manganese horseradish peroxidase was thus concluded to be Mn(IV), S = 3/2. Mn(III)-cytochrome-c peroxidase reacted with stoichiometry quantities of hydrogen peroxide to form a catalytically active intermediate. The electronic absorption spectrum was very similar to that of a higher oxidation state of manganese porphyrin, Mn(V). Since the peroxide compound of manganese cytochrome-c peroxidase retained two oxidizing equivalents per mol of the enzyme (Yonetani, T. and Asakura, T. (1969) J. Biol. Chem. 244, 4580-4588), this peroxide compound might contain an Mn(V) center. PMID- 3030432 TI - Amino acid sequences of the two heme c-binding sites of Pseudomonas cytochrome-c peroxidase. AB - The amino acid sequences of the two heme c-containing tryptic peptides of Pseudomonas cytochrome-c peroxidase have been determined. The tryptic peptides were isolated from two cyanogen bromide fragments of the protein. Both heme binding sites have the Cys-X-Y-Cys-His structure characteristic of c-type cytochromes. The sequences of the two peptides show distinct homology with each other, suggesting the occurrence of gene doubling during evolution of the protein molecule. The function of the heme c moieties in the catalytic cycle of the enzyme is discussed on the basis of their homology with the proximal histidine region of peroxidase (horseradish peroxidase and yeast cytochrome-c peroxidase) and cytochromes (horse cytochrome c and Pseudomonas cytochrome c-551). PMID- 3030433 TI - A proton NMR study of the non-covalent complex of horse cytochrome c and yeast cytochrome-c peroxidase and its comparison with other interacting protein complexes. AB - Cytochrome-c peroxidase (ferrocytochrome-c:hydrogen-peroxide oxidoreductase, EC 1.11.1.5) forms a noncovalent 1:1 complex with horse cytochrome c in low ionic strength solution that is detectable by proton NMR spectroscopy. When the entire proton hyperfine-shifted spectrum is considered only five hyperfine resonances exhibit unambiguously detectable shifts: the heme 8-CH3 and 3-CH3 resonances, single proton resonances near 19 ppm and -4 ppm and the methionine-80 methyl group. These shifts are very similar to those observed for the covalently crosslinked complex of cytochrome-c peroxidase and horse cytochrome c, but different from those reported for cytochrome c complexes with flavodoxin and cytochrome b5. By comparison with the shifts reported for lysine-13-modified cytochrome c we conclude that the results reported here support the Poulos-Kraut proposed structure for the molecular redox complex between cytochrome-c peroxidase and cytochrome c. These results indicate that the principal site of interaction with cytochrome-c peroxidase is the exposed heme edge of horse cytochrome c, in proximity to lysine-13 and the heme pyrrole II. The noncovalent cytochrome-c peroxidase-cytochrome c complex exists in the rapid-exchange time limit even at 500 mHz proton frequency. Our data provide an improved estimate of the minimum off-rate for exchanging cytochrome c as 1133 (+/- 120) s-1 at 23 degrees C. PMID- 3030434 TI - Molecular cloning and sequence analysis of the (Na+ + K+)-ATPase beta subunit from dog kidney. AB - cDNA complementary to mRNA coding for the beta subunit of dog renal (Na+ + K+) ATPase has been cloned into lambda gt11 and the nucleotide sequence of the DNA has been determined. The amino acid sequence of the beta subunit polypeptide has also been deduced from the DNA. The mature form of the dog kidney beta subunit contains 302 amino acids with three potential asparagine-linked attachment sites for carbohydrate. The initiation methionine is removed during processing of the polypeptide to its mature form. Although the beta subunit is an integral membrane protein there is no signal sequence for the polypeptide, and hydropathy analysis predicts that the beta subunit polypeptide spans the cell membrane only once. Secondary structure predictions and a model for the structure of the beta subunit are proposed. DNA sequencing of the 5' non-coding region of the mRNA revealed a 200 bp inverted repeat from the coding region. Blot hybridization of a fragment of the beta subunit cDNA identified a single mRNA species of 2.7 kb in dog kidney and several rat tissues. RNA from rat liver was deficient in mRNA that hybridized to the dog kidney beta subunit cDNA, although mRNA that hybridized to an alpha subunit cDNA was detected. RNA from a human hepatoma cell line, HepG2, however, contained comparable levels of mRNA for both the alpha and the beta subunits. PMID- 3030435 TI - Isolation, purification and 1H-NMR characterization of a kringle 5 domain fragment from human plasminogen. AB - A scheme is proposed for generating the intact Val-448-Phe-545 polypeptide of human plasminogen which contains the fifth kringle domain of the plasmin heavy chain. The procedure is based on a pepsin fragmentation of miniplasminogen and involves the purification of the kringle 5-containing fragment by gel filtration and ion-exchange chromatography. The final product is characterized by amino acid analysis, N- and C-terminal analyses, and high-resolution 1H-NMR spectroscopy at both 300 MHz and 611 MHz. We detect a (40:60%) Asp/Asn heterogeneity at site 452 of the Glu-plasminogen molecule. In the conventional kringle numbering system, the kringle 5 domain extends from Cys-1 to Cys-80, which corresponds to Cys-461 to Cys-540 in plasminogen. A preliminary 1H-NMR characterization of kringle 5 focuses on the global conformational features of the polypeptide. Assignments are given for a number of resonances, including the Tyr-72, the His imidazoles' and the Trp indoles' spin systems. Comparison with human plasminogen kringles 1 and 4 shows that the kringle 5 conformation is highly structured and very similar to that of the homologous domains. This conservancy is particularly striking in the environment surrounding Leu-46 and in the overall features of the aromatic spectrum. There are some differences, particularly in the buried His-33 imidazole group, whose H2 resonance is shifted to 9.67 ppm. A preliminary study of benzamidine-binding shows that the ligand interacts weakly (Ka approximately equal to 1.7 mM -1) mainly through the amidino functional group. Trp-62 and Tyr 72 are significantly perturbed by benzamidine, suggesting that these residues are part of the ligand-binding site. PMID- 3030436 TI - Calpain abolishes the effect of filamin on the actomyosin system in platelets. AB - Platelet filamin was shown to cross-link F-actin and inhibit actomyosin ATPase activity. Filamin was also shown to be degraded by calpain (calcium-activated neutral proteinase; CANP) when the platelet was activated. The consequences of the proteolysis of filamin on the actomyosin system have been investigated. When degraded by calpain in the presence of Ca2+, filamin loses its ability to cross link F-actin. Under the same conditions, its inhibitory effects on the superprecipitation and ATPase activity of actomyosin are abolished. The result suggests that the degradation of filamin is favorable for contraction of the activated platelets. PMID- 3030437 TI - Partial purification of ethanolaminephosphotransferase from rat brain microsomes. AB - Rat brain ethanolaminephosphotransferase (CDPethanolamine : 1,2-diacylglycerol ethanolaminephosphotransferase, EC 2.7.8.1) was solubilized by treating rat brain microsomes with buffered solutions containing octyl glucoside or Triton X-100. The solubilized enzyme was stable both at 4 degrees C and at -18 degrees C. A partial purification was obtained using an ion-exchange chromatographic procedure. The partially purified enzyme showed four major bands in SDS polyacrylamide gel electrophoresis; its specific activity was increased by a factor of 37 compared to that of the membrane-bound enzyme. Glycerol and diacylglycerol were effective as stabilizers. Phosphatidylcholine, lysophosphatidylcholine and phosphatidylserine increased both the specific activity and the stability of the partially purified enzyme. PMID- 3030438 TI - Acyl-chain specificity and properties of cholesterol esterases from normal and Wolman lymphoid cell lines. AB - Cholesteryl esters with various chain lengths of fatty acid, radioactive (C2 C18:1) and fluorescent (pyrene butanoic and decanoic acid, P4 and P10, respectively) were synthesized and their hydrolysis was investigated in lymphoid cell lines from normal subjects and from Wolman's disease patients. The comparison of their hydrolysis showed that three cholesterol esterases were present in normal lymphoid cell lines: the first, active at pH 4.0, hydrolysed preferentially cholesteryl esters of acyl chain length more than 8 carbons, and P10-cholesteryl ester. This acid cholesterol esterase, strongly inhibited by SH blocking agents and resistant to E600, was severely deficient in Wolman lymphoid cell lines and corresponded to acid lysosomal lipase. The second and the third cholesterol esterases, active at pH 6.0 and 8.0, respectively, hydrolysed shorter chain derivatives: the pH 8.0 enzyme was specific for short-chain derivatives (cholesteryl acetate, butyrate and P4), whereas the pH 6.0 activity showed a broader specificity, since it hydrolysed all the cholesteryl esters, with a maximum of activity on cholesteryl acetate and butyrate. The pH 6.0 and 8.0 enzymes were heat-labile, inhibited by E600, resistant to SH-blocking agents and not deficient in Wolman lymphoid cell lines. The hypothetical physiological role of these enzymes is discussed. PMID- 3030439 TI - Selective inhibition of 5-lipoxygenase pathway in rat pulmonary alveolar macrophages by cigarette smoking. AB - Pulmonary alveolar macrophages from sham or cigarette-smoke-exposed rats were examined for their ability to transform exogenously added arachidonate to metabolites of lipoxygenase and cyclooxygenase pathways. Synthesis of 5-HETE and leukotriene B4 was selectively inhibited by cigarette smoke exposure, whereas the formation of prostaglandin E2 and thromboxane B2 remained unchanged. Selective inhibition of the lipoxygenase pathway was further reflected by the reduced content of leukotriene B4 in bronchoalveolar fluid of smoke-exposed rats. These results suggest that lipoxygenase-derived products may play a unique role in smoking-induced pulmonary diseases. PMID- 3030440 TI - Short-term metabolism of cholesteryl ester from low-density lipoprotein in primary monolayers of bovine adrenal cortical cells. AB - The uptake and metabolism of [14C]cholesteryl ester in bovine LDL to cortisol and to cholesteryl ester was studied in monolayer cultures of bovine adrenal cortical cells over short time periods of up to 8 h. The experiments were designed to determine the intracellular pathway followed by the cholesterol derived from the LDL cholesteryl ester and how this is modified in the short term by the tropic hormone ACTH. The cells were cultured in the presence of mevinolin to remove the contribution of endogenous synthesis of cholesterol for supply of substrate for steroidogenesis. The specific activity of the cortisol secreted by the cells was measured under a variety of conditions. Control incubations showed a relatively steady specific activity in the cortisol secreted over an 8 h period. In the presence of ACTH the specific activity of the cortisol was significantly reduced for the first 2 h of the experiment. This is consistent with dilution of the [14C]cholesterol from the LDL with non-radioactive free cholesterol released from the intracellular stores of cholesteryl ester in the presence of ACTH. The inhibitor of acyl-CoA:cholesterol acyltransferase, Sandoz compound 58-035, increased the specific activity of the secreted cortisol in the absence of ACTH, indicating that much of the incoming cholesterol would normally be esterified but was here diverted to steroidogenesis. In the presence of ACTH this increase was observed only during the first 2 h of the experiment, after which inhibition of acyl-CoA:cholesterol acyltransferase had no effect on the specific activity of the cortisol. The adrenal cells were further fractionated into mitochondrial, lysosomal and microsomal plus cytosol fractions and the appearance of free and esterified cholesterol from the labelled LDL measured in these fractions over a period of up to 8 h. ACTH stimulated the uptake of LDL-cholesteryl ester into the cells and tended to increase the relative amounts of free cholesterol in the cells, consistent with its role in promoting supply of cholesterol for steroidogenesis. These experiments allow the roles of endogenous cholesteryl ester and lipoprotein-derived cholesteryl ester in the bovine adrenal cortical cells to be observed over a short time scale. They show that the cells make a substantial change in the internal flux of cholesterol in a short time after stimulation with ACTH and in these cultures the full expression of the presence of ACTH takes up to 2 h. PMID- 3030441 TI - Superoxide, hydrogen peroxide and singlet oxygen in hematoporphyrin derivative cysteine, -NADH and -light systems. AB - Hematoporphyrin derivative and light in the presence of cysteine or glutathione were found to convert oxygen to superoxide and hydrogen peroxide at pH less than approx. 6.5, while at pH greater than 6.5 no superoxide or hydrogen peroxide production was observed. However, at pH values greater than 6.5 the rate of oxygen consumption increased. This rate paralleled the acid dissociation curve of the cysteine thiol group and is consistent with the chemical quenching of 1O2 by cysteine. The superoxide and hydrogen peroxide formation observed below pH 6.5 appeared not to be related to the singlet oxygen production of hematoporphyrin derivative. In addition, superoxide and hydrogen peroxide production was observed with hematoporphyrin derivative and light in the presence of NADH, both above and below pH 6.5. Direct detection of singlet oxygen luminescence at 1268 nm in the hematoporphyrin derivative-light system (2H2O as solvent) revealed an apparent linear increase in the singlet oxygen emission intensity as the p2H was raised from 7.0 to 10.0. Azide efficiently quenched this observed emission. In addition, at p2H 7.4, 1 mM cysteine resulted in a 40% reduction of the singlet oxygen luminescence, while at p2H 9.4 the signal was quenched by over 95% (under the experimental conditions employed). In total, we interpret these results as consistent with the chemical quenching of 1O2 by the ionized thiol group of cysteine. PMID- 3030442 TI - Oligosaccharides of the Hazelhurst vesicular stomatitis virus glycoprotein are more extensively processed in Rous sarcoma virus-transformed baby hamster kidney cells. AB - Because of the extensive oligosaccharide heterogeneity of the membrane glycoprotein (G) from the Hazelhurst strain of vesicular stomatitis virus, this virus has been used as a specific intracellular probe of altered protein glycosylation in Rous sarcoma virus-transformed versus normal baby hamster kidney cells. Over 70% of G protein from virus released from the transformed cells had acidic-type oligosaccharides at both glycosylation sites, compared to less than 50% from the corresponding normal host cells. The remaining G protein contained an acidic-type oligosaccharide at one site and an endo-beta-N acetylglucosaminidase H-sensitive oligosaccharide at the other. The major endoglycosidase-sensitive species were sialylated hybrid-type (NeuNAc-Gal-GlcNAc Man5GlcNAc2-Asn) from the transformed and neutral-type (Man5-6GlcNAc2-Asn) from the normal host cells. The degree of branching of the acidic-type oligosaccharides was not increased in the transformed cells (approx. 80% biantennary for viral G protein from both cell types). At a reduced growth temperature (24 versus 37 degrees C), the G protein oligosaccharides were more extensively processed in both cell types (approximately 85-95% of G protein contained acidic-type structures at both sites), even though the level of viral protein synthesis and virus release was decreased. Essentially all of the minor, endoglycosidase-sensitive oligosaccharides on mature viral G protein were sialic acid-containing hybrid-type structures. At 24 degrees C the branching of the acidic-type oligosaccharides was increased in the virus released from the transformed cells versus normal cells. PMID- 3030443 TI - The electronic spectra of porphyrin-like cobalt-tetracarboxy-phthalocyanines as modified by polylysine and polyglycols. AB - The effect of poly-L-lysine and polyglycols on the electronic spectral properties of cobalt-tetracarboxy-phthalocyanine, compound (I), was studied in order to determine the effect of the polymers on the molecular stacking properties of compound (I). In the present study we have coupled, both covalently and electrostatically, the poly-L-lysine to compound (I). The electrostatic interaction with the polylysine resulted in a bathochromic shift of absorbance by compound (I) from 680 to 695 nm, and the generation of shoulders in the 590 and 580 nm region. The bathochromic shift indicates that the polylysine either misaligns the units of compound (I) in the molecular stack or alters the angular relationship of the planar compound (I) molecules. The covalent linkage with the polylysine resulted in stabilization of the monomeric form of compound (I) with no hypochromism in the 680 nm region. The reaction of the polyglycols, dextran and DEAE dextran with compound (I) prior to the addition of the polylysine, stabilized the polymeric stacked form of compound (I) in the presence of the polylysine. Ammonium ion and ethanolamine resulted in the appearance of peaks in the 730-760 nm region; a trihydroxy containing primary amine and monomeric lysine generated no such peak at wavelengths above 700 nm. The polyglycol binding capacity of compound (I) facilitated the separation of the unbound compound (I) from the polymeric complexes. PMID- 3030444 TI - Metastatic mouse melanoma cells release collagen-gelatin degrading metalloproteinases as components of shed membrane vesicles. AB - The purpose of this study has been to compare collagen-gelatin degrading enzymes isolated from cancer cell organelles and cytosol to the metalloproteinases released by cancer cells. To this end, metastatic mouse melanoma cell organelles were isolated by sucrose density gradient centrifugation and metalloproteinases were assayed using native and denatured [methyl-3H]collagen substrates. Solubilized proteinases were purified by ammonium sulfate precipitation, anion exchange, concanavalin A affinity and gel-filtration column chromatographic procedures and characterized by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The conclusions were as follows: malignant melanoma cells have a metalloproteinase (Mr = 59,000) which is shed from cells into conditioned medium as a component of intact membrane vesicles rather than as a soluble enzyme; storage of tumor-conditioned medium leads to the generation of autoactivated soluble metalloproteinases of lower molecular weight; purification of these metalloproteinase species yielded variant collagenases that have considerable gelatinolytic activity and a cleavage preference site for the Gly-Ile bond in a collagen-like synthetic octapeptide substrate which is typical for collagenase type metalloproteinases. It is proposed that localization of potent proteinases to the surface of cancer cells facilitates the local breakdown of connective tissues during the invasive process. PMID- 3030445 TI - A vertebrate interstitial collagenase inhibitor from bovine scapular cartilage: purification and characterization. AB - A collagenase inhibitor was purified from bovine cartilage by a combination of gel filtration, ion exchange, concanavalin A-Sepharose affinity chromatography, and elution from preparative sodium dodecyl sulfate-polyacrylamide gels. The inhibitor was purified 370-fold and migrated as a single polypeptide with an Mr of 19,000 on SDS-polyacrylamide gels. It stained positively for carbohydrate with periodic acid-Schiff's reagent and bound to lectins, indicating that it is a glycoprotein. The inhibitory activity was stable to heating up to 60 degrees C and between pH 4 and 10. The inhibition of collagenase by the cartilage inhibitor could not be reversed by trypsin or mersalyl. The inhibitory activity did not require the presence of free sulfhydryl groups, and it could be removed from the cartilage extract by incubating with native collagen, suggesting that the inhibitor binds to collagen. The cartilage inhibitor was effective against human and mouse interstitial collagenases, but it did not inhibit trypsin or bacterial collagenase. PMID- 3030446 TI - Collagen-binding proteins secreted by type II pneumocytes in culture. AB - The alveolar epithelial basement membrane is believed to play important roles in lung development, in maintaining normal alveolar architecture, and in guiding repair following lung injury. However, little is known about the formation of this structure, or of the mechanisms which mediate interactions between the epithelium and specific matrix macromolecules. Since type IV collagen is a major structural component of basement membranes, we investigated the production of type IV collagen-binding proteins by primary cultures of rat lung type II pneumocytes. Cultures were labeled for up to 24 h with 3H-labeled amino acids or [3H]mannose. Soluble collagen-binding proteins which accumulated in the culture medium were isolated by chromatography on collagen-Sepharose and examined by SDS polyacrylamide gel electrophoresis. The major type IV collagen-binding protein (CBP1) was identified as fibronectin. We also identified a novel disulfide-bonded collagen-binding glycoprotein (CBP2; Mr = 45,000, reduced). This protein was not recognized by polyclonal antibodies to fibronectin, and showed no detectable binding to denatured type I collagen. The protein was resolved from fibronectin and partially purified by sequential chromatography on gelatin and type IV collagen-Sepharose. We suggest that type II pneumocyte-derived collagen-binding proteins contribute to the formation of the epithelial basement membrane and/or mediate the attachment of these cells to collagenous components of the extracellular matrix. PMID- 3030447 TI - Diffusion of singlet oxygen into human bronchial epithelial cells. AB - The respiratory epithelium undergoes morphological and functional changes following exposure to single oxygen. However, mechanisms by which singlet oxygen causes cellular injury are unclear. The present experiments were designed to investigate the possibility that singlet oxygen, a highly reactive species, diffuses into respiratory epithelial cells. Of the various methods for detection of singlet oxygen, the electron spin resonance (ESR) spectrometric technique was judged to be most compatible and sensitive for use with cell suspensions. ESR spectrometry was used to monitor the singlet oxygen-mediated conversion of 2 (9,10-dimethoxyanthracenyl)-tert-butylhydroxylamine, (I), to 2-(9,10 dimethoxyanthracenyl)-tert-butylnitroxide, (II), and its corresponding endoperoxide, (III), in human bronchial epithelial cells treated with extracellularly generated singlet oxygen. In a second series of experiments, bronchial epithelial cells labeled with (I) were treated with singlet oxygen in the presence of 1,4-diazabicyclo[2.2.2]octane, a singlet oxygen quenching agent. The addition of this quenching agent eliminated the ESR spectrum corresponding with (II) and (III). This result is consistent with the quenching of singlet oxygen by 1.4-diazabicyclo[2.2.2]octane. Collectively, our results indicate that extracellularly generated singlet oxygen diffuses into human bronchial epithelial cells and that this process is a potentially important step in the cytotoxic action of singlet oxygen to the respiratory epithelium. PMID- 3030449 TI - Possible role of a cAMP-dependent phosphorylation in the calcium release mediated by inositol 1,4,5-trisphosphate in human platelet membrane vesicles. AB - The addition of inositol 1,4,5-trisphosphate (IP3) to a 45Ca-preloaded human platelet membrane fraction (dense tubular system) induced a transient release of Ca2+. When the vesicle fraction was loaded with 45Ca2+ to isotopic equilibrium in the presence of the catalytic subunit of the cAMP-dependent protein kinase, the level of Ca2+ uptake was increased and the subsequent IP3-induced Ca2+ release was enhanced. The stimulation was observed regardless of the IP3 concentration used, and was maximal with an enzyme concentration of 5 micrograms/ml. The addition of the protein kinase inhibitor prevented the stimulatory effect of the catalytic subunit on IP3-induced calcium release, and also abolished the calcium release detected in the absence of added enzyme. It is concluded that a cAMP dependent protein phosphorylation may be involved in the regulation of the IP3 induced Ca2+ release in human platelets. PMID- 3030448 TI - Effects of phosphorylation of protein phosphatase 1 by pp60v-src on the interaction of the enzyme with substrates and inhibitor proteins. AB - Phosphorylation of protein phosphatase 1 by pp60v-src decreased its activity towards phosphorylase kinase and glycogen synthase as well as towards phosphorylase a. Kinetic experiments indicated that the primary effect of phosphorylation was to increase the Km for each of the substrate proteins. There was little or no change in the Vmax for the reactions. The possibility that phosphorylation of protein phosphatase 1 altered its regulation by inhibitors-1 and -2 was also examined. Phosphorylation of protein phosphatase 1 did not prevent the reversible inhibition of the enzyme by inhibitor-1 or inhibitor-2 nor did it prevent the association of inhibitor-2 with protein phosphatase 1 to form the MgATP-dependent protein phosphatase. Protein phosphatase 1 is not a substrate for pp60v-src when it is complexed with inhibitor-2 to form the inactive MgATP dependent protein phosphatase. Here we have shown that protein phosphatase 1 is also not phosphorylated by pp60v-src following activation of the MgATP-dependent protein phosphatase with glycogen synthase kinase-3 and MgATP. This indicates that the inability of pp60v-src to phosphorylate protein phosphatase 1 is not due to the change in protein phosphatase 1 conformation which accompanies the inactivation of the MgATP-dependent protein phosphatase. Rather, it appears to be the result of steric hindrance by inhibitor-2. This suggests that the pp60v-src phosphorylation site is closely associated with the inhibitor-2 binding site involved in the formation of the MgATP dependent protein phosphatase. The pp60v src phosphorylation site was previously localized to a small (Mr less than or equal to 4000) domain which can be selectively degraded by chymotrypsin. Here we have shown that chymotryptic digestion increased the Km of unphosphorylated protein phosphatase 1 for each of the three phosphoprotein substrates used in this study. This effect was similar to that observed after phosphorylation of protein phosphatase 1. These results indicate that the pp60v-src phosphorylation site is in a region of protein phosphatase 1 which influences substrate binding and which may be near the active site. PMID- 3030450 TI - Studies on the mechanism of decreased NMR-measured free magnesium in stored erythrocytes. AB - 31P-NMR spectra have been recorded on erythrocytes stored at 4 degrees C in various preservation media. Storage was always associated with an upfield shift of the inorganic phosphate (Pi) resonance and a pronounced upfield shift of the ATP beta resonance, indicating decreased intracellular pH (pHi) and decreased intracellular free magnesium ([Mg2+]i). The decreased [Mg2+]i occurred in preservation media not containing citrate and even in media supplemented with Mg2+. It could not be attributed to the changes in pHi, Na+, K+, lactate, Pi or 2,3-diphosphoglycerate, that occur with storage. The decrease in [Mg2+]i was largely reversed when stored cells were incubated for 1 h at 37 degrees C in fresh plasma. Stored cells were found to contain significant amounts of inorganic pyrophosphate, up to about 200 mumol per liter cell water. Being a tight binder of Mg2+, pyrophosphate could account for some of the observed decrease in [Mg2+]i. Additional mechanisms may involve precipitation of some other Mg2+ complex during cold storage or enhancement of Mg2+ binding to membrane components. PMID- 3030451 TI - Phosphorylation of rat heart glycogen synthase: studies in cardiomyocytes and in vitro phosphorylations with cAMP-dependent kinase and protein phosphatase-1. AB - The phosphorylation of glycogen synthase has been studied in freshly isolated adult rat cardiomyocytes. Six peaks of 32P-labeled tryptic peptides are recovered via C-18 high performance liquid chromatography (HPLC) when synthase is immunoprecipitated from 32P-labeled cardiomyocytes and digested with trypsin. When epinephrine treated cells are used as a source of enzyme, the same HPLC profile is obtained with a dramatic enhancement of 32P recovered in two of the HPLC peaks. In vitro phosphorylation of rat heart synthase by cAMP-dependent protein kinase stimulates the conversion of synthase from the I to the D form and results in the recovery of the same tryptic peptides from the C-18 as is the case for synthase derived from cardiomyocytes. Treatment of cAMP-dependent kinase phosphorylated synthase with protein phosphatase-1 leads to a reactivation of the enzyme and a dephosphorylation of the same tryptic peptides that are selectively phosphorylated in epinephrine treated cardiomyocytes. These results are discussed in relation to hormonal control of glycogen metabolism in cardiac tissue. PMID- 3030452 TI - [Interaction of alpha-tocopherol with free fatty acids. Mechanism of stabilization of lipid bilayer microviscosity]. AB - Using ESR-spin probes and 1H-NMR-spectroscopy methods the effect of alpha tocopherol on liposome microviscosity has been studied. alpha-Tocopherol has been shown to remove the chaotropic action of free fatty acids on bilayer. The stabilization effect found has a common nature and does not depend on the chemical structure of the phopsholipid functional polar groups, the unsaturation degree of free fatty acids as well as fatty acids residua entering into phospholipid composition. Analog of alpha-tocopherol without phytol chain 2,2,5,7,8-penthamethyl-6-oxychroman does not show the stabilizing effect on the microviscosity of lipid bilayer under the action of free fatty acids. It indicates that both chromanol nucleus and phytol chain of alpha-tocopherol molecule are necessary for stabilizing action. The data obtained allow to suppose that the interaction of alpha-tocopherol with free fatty acids may be one of the molecular mechanisms of lipid bilayer microvicosity stabilization. PMID- 3030453 TI - [Changes in the structure of interphase nuclei of lymph node stromal cells after treatment with cAMP]. AB - The exogenic cAMP has been shown to induce the extrusion of nucleoli and some of nucleic material into cytoplasma of reticular cells of mouse lymph nodes. PMID- 3030454 TI - Interactive roles of progesterone, prostaglandins, and collagenase in the ovulatory mechanism of the ewe. AB - Interrelationships between production of progesterone (P4), prostaglandin (PG) E2 and PGF2 alpha, and collagenase by periovulatory ovine follicles and their possible involvements in the ovulatory process were investigated. Follicles were isolated from ovaries at intervals (0 to 24 h) after the initiation of the preovulatory surge of luteinizing hormone (LH). Progesterone and PGs within follicles were determined by radioimmunoassay. Digestion of radioactive collagen during coincubation with tissue homogenates was used to assess the production of a bioactive follicular collagenase(s). Follicular accumulation of PGs and P4 increased at 12 and 16 h, respectively, after the onset of the surge of LH; PGE2 then decreased at 20 h. Collagenolytic activity of follicular tissue increased at 20 h and was maximal at 24 h (during the time of follicular rupture). An inhibitor of synthesis of P4 (isoxazol) or PGs (indomethacin) was injected into the follicular antrum at 8 h. Isoxazol did not prevent the initial rise in PGs, but inhibited synthesis of PGF2 alpha at 16 h and therafter. Isoxazol negated the decline in PGE2 and increase in collagenolysis. Indomethacin did not influence synthesis of P4; however, it suppressed collagenolytic activity of follicular tissue. Ovaries with treated follicles were left in situ and observed for an ovulation point at 30 h. Isoxazol or indomethacin was a potent inhibitor of ovulation. The blockade of ovulation by isoxazol was reversed by systemic administration of P4 or PGF2 alpha, but not by PGE2. Reversal of the blockade by indomethacin was accomplished with PGE2 or PGF2 alpha. Collagenolytic activity of follicular tissue was likewise restored by such treatments.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3030455 TI - In vivo leucocyte interactions with the NHLBI-DTB primary reference materials: polyethylene and silica-free polydimethylsiloxane. AB - In vivo leucocyte interactions with the NHLBI-DTB primary reference materials, low density polyethylene (LDPE) and silica-free polydimethylsiloxane (PDMS), were qualitatively and quantitatively characterized using a cage implant system over a 21 d implantation period. Scanning electron microscopy (SEM) and cytochemical staining procedures were utilized to observe the cellular events occurring at the leucocyte/biomaterial interface. The results showed that more cells adhered to the PDMS surface than the LDPE surface at days 4 and 7. The differential analysis revealed that mononuclear cells, presumably macrophages, preferentially adhered to both polymer surfaces. By day 21, there were more very large (greater than 20 nuclei per cell) foreign body giant cells (FBGCs) present on the PDMS surface than the LDPE surface. The phagocytic capabilities of the adhered cells, including the FBGCs, decreased to a greater extent on the PDMS surface, corresponding to the earlier and more extensive spreading of these cells observed in the morphological analysis. PMID- 3030456 TI - Biocompatibility and degradability of sepiolite-collagen complex. AB - Sepiolite, a magnesium silicate, binds collagen resulting in a complex which has a gel-like structure when hydrated. The binding of the protein to the clay decreases its degradability by collagenase, and no degradation was observed after treatment of the complex with glutaraldehyde. Extracts of both the untreated and the glutaraldehyde-treated complex are biocompatible for fibroblast growth. Based on its properties, this material should be considered in the design of biomaterials. PMID- 3030457 TI - Cytochrome c binds to lipid domains in arrays of mitochondrial outer membrane channels. AB - Computer-averaged electron microscopic images of negatively stained crystalline arrays of fungal mitochondrial outer-membrane channels in the presence and absence of cytochrome c were compared. Neither the apo- nor the holo- forms of cytochrome c significantly changed the stain distribution in the protein regions of the channel arrays. However, both forms of cytochrome c caused significant stain exclusion from the lipid domains in the arrays, suggesting binding of the polypeptides at these loci. The implications of binding of apocytochrome c to clusters of exposed phospholipids on the mitochondrial outer membrane are discussed with respect to the mechanism of uptake of this polypeptide by mitochondria. PMID- 3030459 TI - Interaction of quercetin with DNA. AB - In vitro experiments to study interaction of the mutagenic flavonoid quercetin with DNA are described. Calf thymus DNA treated with quercetin for various time periods was subjected to S1 nuclease hydrolysis. Thermal melting profiles of treated DNA were also determined using S1 nuclease. The rate of DNA hydrolyzed after 1 hr of pretreatment with quercetin was found to be only about 50% of that in its absence. However, after 10 and 24 hrs of treatment with the drug, the rate of S1 nuclease hydrolysis was observed to be greater than that of native DNA. Thermal melting profiles of DNA, treated with quercetin for 10 and 24 hrs, indicated a slight decrease in melting temperatures. Gel filtration of native DNA, which had been digested with S1 nuclease after preincubation with quercetin for 24 hrs, indicated the production of various sized degraded molecules. The results suggest that the initial interaction of quercetin with DNA may have a stabilizing effect on its secondary structure, but prolonged treatment leads to an extensive disruption of the double helix. PMID- 3030458 TI - Orientation of spin-labeled nucleotides bound to myosin in glycerinated muscle fibers. AB - Electron paramagnetic resonance (EPR) spectroscopy of paramagnetic derivatives of ATP has been used to probe the angular distribution of myosin in glycerinated muscle fibers. Three nucleotide spin labels have been prepared with the nitroxide free radical moiety attached, via an ester linkage to either: the 2' or 3' positions of the ribose unit of ATP (SL-ATP), the 2' position of 3' deoxy ATP (2'SL-dATP), or the 3' position of 2' deoxy ATP (3'SL-dATP). In muscle fibers, these nucleotides are quickly hydrolyzed to their diphosphate forms. All three diphosphate analogues bind to the nucleotide site of myosin with similar affinities: rabbit psoas fibers, 7 X 10(3)/M; insect flight muscle, 5 X 10(3)/M; and rabbit soleus muscle, 2 X 10(4)/M. Analysis of the spectra showed that the principal z-axis of the nitroxide attached to bound nucleotides was oriented with respect to the filament axis. The principal axes of 3'SL-dADP and 2'SL-dADP appeared to be preferentially aligned at mean angles of 67 degrees +/- 4 degrees and 55 degrees +/- 5 degrees, respectively. The distribution of probes about these angles can be described by Gaussians with widths of 16 degrees +/- 4 degrees and 13 degrees +/- 5 degrees, respectively. The spectrum of bound SL-ADP was a linear combination of the spectra of the two deoxy analogues. These orientations were the same in the three muscle types examined, indicating a high degree of homology in the nucleotide binding site. Applying static strains as high as 0.2 N/mm2 to muscle fibers caused no change in the orientation of myosin bound, spin-labeled nucleotides. When muscle fibers were stretched to decrease actin and myosin filament overlap, bound SL-ADP produced EPR spectra indicative of probes with a highly disordered angular distribution. Sodium vanadate and SL ATP caused fiber stiffness to decrease, and the EPR spectrum of the bound analogue indicated an increase in the fraction of disoriented probes with a concomitant decrease in the fraction of oriented probes. These findings indicate that when myosin is bound to actin its nucleotide site is highly oriented relative to the fiber axis, and when this interaction is removed the orientation of the nucleotide site becomes highly disordered. PMID- 3030460 TI - [Purification and study of the physicochemical properties of angiotensin converting enzyme from the human liver]. AB - Angiotensin-converting enzyme (ACE) from human liver was first purified 9000-fold by chromatofocusing with 22% yield. The enzyme had a specific activity of 10 U/mg. The enzyme molecular weight was 150000, as determined by electrophoresis in a 7.5% polyacrylamide gel. The enzyme pI determined by chromatofocusing was 4.2 4.3. KM of human liver ACE, measured using hippuryl-L-histidyl-L-leucine and N benzyloxycarbonyl-L-phenylalanyl-L-histidyl-L-leucine as substrates, was 5 mM and 0.1 mM, respectively. Human liver ACE was inhibited by SQ 20881 with IC50 equal to 1.8 X 10(-8) M. PMID- 3030461 TI - [Analysis of the effect of immunostimulants on the macrophages of mouse peritoneal exudates by using mathematical simulation and planning methods]. AB - Using methods of mathematical modelling and planning an analysis was made of changes in 5-nucleotidase activity in murine peritoneal macrophages after intraperitoneal administration of immunostimulants of different chemical structure and biological origin. The changes in 5-nucleotidase activity after the administration of immunostimulants were shown to exhibit a similar linear pattern. PMID- 3030462 TI - [Functional properties of E. coli SSB-proteins in vivo]. AB - An essential function of single-stranded DNA-binding (SSB) proteins--a defense from nuclease action, determined by their first and second domains, is realized in conditions of normal cell metabolism. With the inhibition of the replicative furcula growth and total protein synthesis (imbalance state), the SSB protein function associated with the third domain and responsible for certain modifications in DNA polymerase II activity is realized. This causes DNA degeneration and increases radiosensitivity of cells. PMID- 3030463 TI - [A protein related to the main core protein of the mouse mammary cancer virus in a microparticle fraction of human milk]. AB - Virus-like density fractions (VDF) have been prepared from 6 specimens of human milk by ultracentrifugation of milk sera and recentrifugation of pellet through 35% sucrose or by separation in sucrose density gradient. Using electroblotting and enzyme-linked antibody probes, the protein with a molecular weight of 27 kD, reacting with serum against MMTV gag proteins, was identified in 5/6 of milk VDF. The absence (or very low amounts) of this protein among proteins of milk, fat globule membranes and cellular membranes of human milk has been demonstrated. Positive VDF decrease the activity of anti-p27 MMTV-specific serum and the activity of human sera reacting with p27 MMTV. A human protein immunologically related to the main gag protein of MMTV has been described; this protein seems to be an antigen causing the appearance of anti-MMTV antibodies in human sera. PMID- 3030465 TI - A component of factor VIII preparations which can be separated from factor VIII activity down modulates human monocyte functions. AB - In this study we investigated different aspects of monocyte functions following interaction of monocytes (Mo) with therapeutic concentrations of factor VIII (F VIII) concentrate. A short (one-hour) treatment of normal Mo with F VIII concentrates led to a significant (P less than 0.001) down modulation of Fc receptors expressed in the Mo plasma membrane. This down modulation was accompanied by a decrease of Mo effector functions that was expressed by a reduced capacity of F VIII-treated Mo to release O2 radicals (40% of controls) and to kill bacteria (% killing: control Mo, 65%; F VIII-treated Mo, 24% to 51%). Further studies showed that the modulating activity was due to a contaminant present in F VIII concentrates (immune complexes or IgG aggregates). Fractionation using molecular sieving revealed that the modulatory activity was confined to a high-molecular range fraction (Mr greater than 1,270,000 daltons), while the fraction containing monomeric IgG had no effect. Further fractionation by affinity chromatography on protein A-Sepharose separated the coagulation activity (effluent) from the Mo function-modulating activity (eluate). We conclude that treatment with F VIII concentrates might contribute to an immunocompromised state in some hemophiliacs and facilitate opportunistic infections in these patients. PMID- 3030464 TI - Contrasting patterns of DNA strand breakage and ADP-ribosylation-dependent DNA ligation during granulocyte and monocyte differentiation. AB - Previous studies have shown that structural changes in DNA, including the ligation of pre-existing DNA breaks and the opening and closure of new breaks, occur shortly after exposure of granulomonocytic precursors (CFU-GM) to granulocyte-macrophage colony stimulating activity (GM-CSA). Monocytic differentiation of CFU-GM is selectively inhibited by compounds known to inhibit the nuclear enzyme ADP-ribosyl transferase (ADPRT). Since this enzyme, which transfers ADP-ribose units to chromatin proteins, is known to activate DNA ligase, we attempted to determine whether ligation of one or both types of DNA break is required for monocytic differentiation. Breaks in DNA were examined using the nucleoid sedimentation technique in which DNA breaks cause loss of DNA supercoiling in nucleoids and concomitant changes in their sedimentation through neutral sucrose gradients. We here report that two distinct patterns of DNA strand breakage and ligation are associated with differentiation to the granulocyte and monocyte lineages. Monocytic inducers (phorbolester and vitamin D3) predominantly produce closure of pre-existing strand breaks, whereas granulocytic inducers (granulocyte colony stimulating activity, G-CSA; retinoic acid) cause opening and closure of new breaks. Only ligation of the pre-existing breaks is highly sensitive to inhibition by 3-methoxybenzamide (a potent ADPRT inhibitor), and only monocytic differentiation is impaired by addition of this compound. These findings suggest that DNA structural changes may be directly involved in granulocyte-macrophage switching. PMID- 3030466 TI - Prevention of pulmonary injury in isolated perfused rat lungs by activated human neutrophils preincubated with anti-Mo1 monoclonal antibody. AB - Neutrophil activation results in neutrophil adherence and may subsequently cause lung injury through the generation of oxidants, release of granule proteases, and generation of a variety of mediator substances. We hypothesized that inhibition of neutrophil adherence and subsequent lung sequestration would attenuate the lung injury caused by activated neutrophils. Using isolated perfused rat lungs, we determined if anti-Mo1 monoclonal antibody (binds to the alpha subunit of a neutrophil glycoprotein [gp 155.94] that facilitates adherence) would attenuate lung neutrophil sequestration and lung injury caused by human neutrophils stimulated by phorbol myristate acetate (PMA). PMA-stimulated neutrophils but not PMA or neutrophils alone caused lung injury as assessed by accumulation of 125I bovine serum albumin into lung parenchyma and alveolar lavage fluid. Incubation of neutrophils with anti-Mo1 antibody prior to stimulation with PMA attenuated lung injury and neutrophil sequestration. Furthermore, a histological survey revealed that anti-Mo1 antibody inhibited neutrophils present in the lung from spreading following exposure to PMA. Anti-Mo1 antibody did not inhibit PMA stimulated neutrophil release of granule constituents or toxic O2 metabolites as evidenced by lysozyme and lactoferrin release or the reduction of ferricytochrome c in the lung perfusate. The inhibition of lung injury caused by the anti-Mo1 antibody was not likely due to a nonspecific effect of the antibody, since another murine monoclonal antibody of the same class (anti-Mo5) did not inhibit lung neutrophil sequestration or lung injury. Thus, in this experimental model, interference with the close approximation of the neutrophil to its target site inhibited the ability of the activated human neutrophil to cause injury. PMID- 3030467 TI - Expression of the macrophage-specific colony-stimulating factor in human monocytes treated with granulocyte-macrophage colony-stimulating factor. AB - The macrophage-specific colony-stimulating factor (CSF-1, M-CSF) regulates the survival, growth and differentiation of monocytes. We have recently demonstrated that phorbol ester induces expression of CSF-1 in human monocytes. These findings suggested that activated monocytes are capable of producing their own lineage specific CSF. The present studies demonstrate that the granulocyte-macrophage colony-stimulating factor (GM-CSF) also induces CSF-1 transcripts in monocytes. Furthermore, we demonstrate that the detection of CSF-1 RNA in GM-CSF-treated monocytes is associated with synthesis of the CSF-1 gene product. The results thus suggest that GM-CSF may indirectly control specific monocyte functions through the regulation of CSF-1 production. These findings indicate another level of interaction between T cells and monocytes. PMID- 3030468 TI - Unproportionally high concentrations of diadenosine triphosphate (Ap3A) and diadenosine tetraphosphate (Ap4A) in heavy platelets. Consequences for in vitro studies with human platelets. AB - Platelets from whole blood were separated into five density subpopulations using a discontinuous Percoll gradient. The content of diadenosine triphosphate (Ap3A), diadenosine tetraphosphate (Ap4A), ADP and ATP were determined in the subfractions. The dinucleotides were directly measured in neutralized, acid soluble extracts of human platelets with a bioluminescence method not requiring any chromatographic step. When comparing the nucleotide contents of the density subpopulations it became evident that all nucleotides steadily increased with increasing density. Ap3A, Ap4A, ADP and ATP were present in 10-, 7-, 4- and 2 fold higher amounts in the heaviest platelets, respectively, as compared to the subfraction with the lowest density. This finding is practically relevant since the most dense platelet subpopulations may be lost during conventional centrifugation to obtain platelet-rich plasma. Therefore we compared a platelet population obtained from PRP with the platelet population, which had been prepared from whole blood by means of a continuous Percoll gradient. All the four nucleotides investigated were represented in 1.5- to 2-fold higher amounts in the whole blood platelet population. This indicates that PRP does not contain a representative population but lacks part of the large heavy platelets containing the highest amounts of nucleotides. PMID- 3030469 TI - [Experimental vaginitis in rats]. AB - The authors present an experimental model of vaginal inflammation in female rats, through local instillation of different substances causing inflammation of varying degrees. The inflammatory reaction was studied macroscopically, histologically and biochemically. Using polysiloxane as a protective substance of vaginal mucosa was found to be of therapeutic interest. PMID- 3030471 TI - Current concepts in the management of soft tissue sarcomas of the extremities. AB - In recent years the orthopaedist has become the initial physician to see soft tissue neoplasms. A review of the current concepts of treatment of these lesions, therefore, seems to be in order. PMID- 3030470 TI - Differential effects of leukotrienes C4, D4, and E4 in the pulmonary and systemic vasculature of sheep. AB - We investigated the comparative direct and cyclooxygenase-mediated effects of the constituents of slow-reacting substance of anaphylaxis (SRS-A), i.e., leukotriene C4 (LTC4), leukotriene D4 (LTD4) and leukotriene E4 (LTE4) on pulmonary and systemic haemodynamics of sheep. In 20 conscious sheep, measurements of pulmonary vascular resistance (Rpv) and systemic vascular resistance (Rsv) were obtained before and after a rapid intravenous injection of LTC4 (0.1 micrograms X kg-1), LTD4 (0.1 micrograms X kg-1) and LTE4 (1 microgram X kg-1). The same protocol was carried out after pretreatment with the leukotriene antagonist FPL-57231 or the cyclooxygenase inhibitor indomethacin. LTD4 increased mean Rpv to 421% of baseline (p less than 0.001) and had a biphasic effect on mean Rsv, which, following an initial decrease of 18% (p less than 0.05), increased to 143% of baseline (p less than 0.05). LTC4 and LTE4 had no significant effects on Rpv, while they increased mean Rsv to 144% and 143% of baseline, respectively (p less than 0.05). This effect was not preceded by a decrease in Rsv. FPL-57231 completely blocked the effects of LTC4, LTD4 and LTE4 on Rsv, and of LTD4 on Rpv. Indomethacin had no effect on LTC4, LTD4 and LTE4-induced increases in mean Rsv, while it prevented the LTD4-induced initial decrease in mean Rsv. Indomethacin also prevented the LTD4-induced increase in Rpv. A dose-response curve (0.05, 0.1, 0.5 and 1 microgram X kg-1) demonstrated that in raising Rsv, LTE4 was approximately 10 times less potent than LTC4 and LTD4.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3030472 TI - Combined intraoperative and postoperative irradiation for soft tissue sarcomas. AB - A major problem in the management of soft tissue sarcomas is the high rate of local recurrence postoperatively, regardless of whether the surgery performed was radical or conservative. The addition of irradiation to the tumor bed has decreased this risk significantly. In an attempt to achieve maximal local tumor control, a number of patients were treated by means of excisional surgery combined with an intraoperative implant and postoperative external irradiation. A new type of intraoperative irradiation procedure was used, in which iridium (192Ir) is inserted into afterloading nylon catheters which are implanted in the tumor bed at the time that the tumor is surgically excised. PMID- 3030473 TI - Spool-shaped proximal pedal phalanges. AB - We have observed that "spool-shaped" proximal pedal phalanges sometimes are found in single or multiple toes, often affecting both feet. These changes may be present in normal as well as abnormal feet. The etiology is unknown. Such changes do not occur in the hands. PMID- 3030474 TI - Clear cell sarcoma of the tendinous portion of the quadriceps muscle at the knee joint managed by limb-sparing surgery. AB - A case report of clear cell sarcoma of the tendinous portion of the quadriceps in the knee joint of a 33-year-old female is presented. This insidious, usually painless malignant tumor is often misdiagnosed and may be referred to the sports medicine specialist as a sport-related injury. The duration of symptoms and the size of the lesion are important prognostic factors. Wide excision or radical surgery is necessary to cure such tumors. With improved reconstructive techniques, including free vascularized myocutaneous flaps, limb-sparing surgery is now possible. PMID- 3030475 TI - Carpal tunnel syndrome secondary to tuberculous tenosynovitis. AB - A case of tuberculous flexor tenosynovitis causing a carpal tunnel syndrome is described. Surgery initially relieved the patient's symptoms, but the disease process continued until the diagnosis was made and appropriate treatment was instituted. This diagnosis, although rare, should be considered as the etiology in a carpal tunnel syndrome, particularly when there is evidence of a proliferative tenosynovitis and radiographs show diffuse osteoporosis. PMID- 3030476 TI - Analogue femurs for laboratory investigations of prosthesis stress distributions and fixation failures. AB - A polymeric analogue femur in combination with a brittle-coating stress-analysis technique was used to compare the relative femoral surface-strain distributions for various hip prostheses. The influence of prosthesis design factors, such as the geometry and material, the effect of a collar, and surface texturing, were readily ascertained and in agreement with previous investigations using other techniques. The fixation failures observed with this laboratory test model were similar to those of published clinical findings. PMID- 3030477 TI - An easy method for the diagnosis of acetabular fractures. AB - An accurate diagnosis of acetabular fractures requires both an anteroposterior radiograph, and two oblique views which are usually obtained by rolling the patient 45 degrees first toward one side and then toward the other. To avoid the discomfort this can cause the patient, we have devised a method that uses standard x-ray equipment but keeps the patient resting supine on the table. The desired oblique views are obtained by tilting the x-ray tube 30 degrees from a medial-to-lateral direction and from a lateral-to-medial direction. PMID- 3030478 TI - Ligament and tendon repair with an absorbable polymer-coated carbon fiber stent. AB - Ribbon-like composite structures of filamentous carbon fiber and absorbable polymers have been used in the repair and replacement of both tendons and ligaments. The composite acts as a scaffold upon which new collagenous tissue can grow and has proved successful in a variety of animal models. The results of the first three years of human clinical trials have revealed ingrowth potential similar to that seen in the animal studies. Most patients have shown significant improvement, with many demonstrating good to excellent stability and function. PMID- 3030479 TI - Mechanical properties and early clinical experience with xenograft biomaterials. AB - Evaluation of new materials for ligament and tendon reconstruction has opened up a new area of orthopaedic research. This report outlines the mechanical properties, animal evaluations, and early clinical results from tests of a xenograft biomaterial of bovine source treated with a gluteraldehyde-based process. A clinical trial under FDA guidelines is currently underway. PMID- 3030480 TI - The effects of the tibial femoral angle and of disuse on the strength of canine anterior cruciate ligaments. AB - Two series of tests were undertaken on dog legs. One series showed that the apparent distraction strength of dog cruciate ligaments depends on the relative angle of the tibia and femur during distraction. The second series documented bone resorption due to disuse. This series showed that mechanical strength was reduced and that there was a reduction in bone collagen due to disuse. PMID- 3030481 TI - Decision-making in low back surgery. AB - The author was the 1986 Sir Robert Jones Lecturer of the Hospital for Joint Diseases Orthopaedic Institute. This article is a slightly edited version of the speech he delivered in that capacity on October 24, 1986, at the Annual Scientific Program of the Alumni Association. PMID- 3030482 TI - Why women are not receiving anti-Rh prophylaxis. PMID- 3030483 TI - Experimental micropolycystic ovarian disease. I. Measurement of body weight, ovarian weight and histochemical activity of 17 beta-hydroxysteroid dehydrogenase. AB - The effects of administration of 1.25 mg testosterone propionate between the 2nd and 5th day of life on body weight, ovarian weight, food and water consumption, and the histochemical activity of 17 beta-hydroxysteroid dehydrogenase (17 beta HSD) were studied in female Wistar rats during the evolution of experimental micropolycystic ovaries. Biometric studies showed a 9.9 day delay of vaginal opening for the treated animals and that the increased body weight of the animals during the disease was not related to food intake and thus presumably due to the induced metabolic disorder. Reduced histochemical activity of the 17 beta-HSD of testosterone-treated rats was detected at ages 30, 60 and 90 days before the onset of morphological alterations of the ovaries. The possible participation of 17 beta-HSD in the disequilibrium between androstenedione and testosterone and in the genesis of the disease is discussed. PMID- 3030484 TI - Leukotrienes and prostaglandins. PMID- 3030486 TI - Vascular responses to leukotriene B4, C4 and D4 following FPL 55712, indomethacin, saralasin, phentolamine and verapamil in the conscious rat. AB - The pressor and vascular permeability effects of leukotrienes B4 (LTB4), C4 and D4 were investigated in conscious unrestrained rats. Leukotrienes C4 and D4 (3.2 51 nmol kg-1 i.v.) caused an acute dose-dependent elevation of the mean arterial pressure, which was maximal after 2 min and returned to control levels within 14 min. Heart rate was significantly reduced by the higher doses of LTC4 and LTD4. LTB4 (up to a dose of 51 nmol kg-1) was essentially inactive. These effects of LTC4 and LTD4 were abolished by FPL 55712, a putative antagonist of sulphidopeptide leukotrienes and by verapamil, a calcium channel blocker. Indomethacin, phentolamine or saralasin pretreatment failed to modify the pressor response to LTC4 and LTD4. LTC4 and LTD4 furthermore caused an increase in haematocrit values, which was significantly attenuated by FPL 55712, indomethacin and verapamil. The present findings show that the pressor effect of LTC4 and LTD4 is not related to prostanoid release and can be reversed by calcium channel blockade; whereas the effect on vascular permeability seems to require the presence of both cyclo-oxygenase product(s) and calcium. PMID- 3030487 TI - Calcium phosphate in catheter encrustation. AB - Encrusted catheters from nine female patients were the source of samples of deposits which were examined by X-ray diffraction, atomic absorption spectroscopy, infra-red spectroscopy and extended X-ray absorption fine structure (EXAFS) spectroscopy. In eight samples the only crystalline phase which could be clearly distinguished by X-ray diffraction was ammonium magnesium orthophosphate hexahydrate, NH4MgPO4 X 6H2O, which occurs naturally as the mineral struvite. However, atomic absorption spectroscopy revealed an appreciable concentration of calcium in all samples. Calcium phosphates have previously been detected in catheter deposits. Infra-red and EXAFS spectra were consistent with the calcium phosphate being present as a poorly crystalline hydroxyapatite. Thus the deposits appear to consist of a mixture of crystalline struvite and a form of hydroxyapatite which is not fully crystalline. PMID- 3030488 TI - Pancreatic endocrine tumours. PMID- 3030485 TI - Crystal-induced inflammation in the rat subcutaneous air-pouch. AB - Monosodium urate (MSU) and calcium pyrophosphate dihydrate (CPPD) crystals initiated acute inflammatory reactions characterized by increased plasma extravasation and polymorphonuclear leukocyte (PMNL) accumulation in the rat subcutaneous air-pouch. Pretreatment of rats with colchicine (1 mg kg-1, s.c.) inhibited PMNL accumulation induced by either crystal type but had a greater inhibitory effect on MSU-induced plasma extravasation compared with that induced by CPPD crystals. Colchicine (1 mg kg-1, s.c.) did not reduce histamine-induced plasma extravasation in the air-pouch. The lipoxygenase product of arachidonic acid metabolism, leukotriene B4 (LTB4), was detected in MSU-induced exudates but not in CPPD-induced exudates. Pretreatment of rats with colchicine (1 mg kg-1, s.c.) inhibited LTB4 production in MSU-induced exudates. PMID- 3030489 TI - A quantitative relationship between Cl- enhanced [3H]flunitrazepam and [35S]t butylbicyclophosphorothionate binding to the benzodiazepine/GABA receptor chloride ionophore complex. AB - A quantitative relationship between the efficacy (i.e. maximum enhancement) of Cl to increase [3H]flunitrazepam binding and the density of [35S]t butylbicyclophosphorothionate binding sites was observed in well-washed membrane fragments of rat cerebral cortex previously exposed to phospholipase A2. This relationship (described by the equation y = ABx) was maintained when [3H]flunitrazepam was assayed at Cl- concentrations between 100 and 600 mM, and was not qualitatively altered by the presence of 100 microM pentobarbital. However, under experimental conditions that reduced the ratio of [35S]TBPS binding sites/benzodiazepine receptors, the effects of pentobarbital suggest that the conductance state of benzodiazepine receptor-coupled chloride channels may be the primary determinant of Cl- enhanced [3H]flunitrazepam binding. PMID- 3030490 TI - Response of supraoptic magnocellular neurons to stimulation of forebrain alpha adrenoceptors. AB - Effects of alpha-adrenoceptor agents on electrophysiologically and immunohistochemically identified supraoptic nucleus (SON) vasopressin (VP) units were investigated by intracarotid infusion. Clonidine, an alpha 2-adrenoceptor agonist always excited SON units and alpha 2-adrenoceptor antagonists consistently inhibited them. alpha 1-Adrenoceptor agents produced inconsistent responses. The results implicate forebrain alpha 2-adrenergic receptors in the excitation of SON VP neurons. PMID- 3030491 TI - Opioid enhancement of perforant path transmission: effect of an enkephalin analog on inhibition and facilitation in the dentate gyrus. AB - The enkephalin analogue [D-Ala2,D-Leu5]enkephalin, applied locally in vivo, enhanced the responsiveness of dentate granule cells of the hippocampus to perforant path stimulation. The facilitatory effect included a decrease in the inhibition and increase in the facilitation induced by sequential stimuli. The results indicate that opioids, acting at opioid receptors, can enhance responsiveness of the dentate gyrus to perforant path transmission, possibly via a disinhibitory mechanism. PMID- 3030492 TI - Intracellular horseradish peroxidase labeling of rapidly firing dorsal raphe projection neurons. AB - The class of rapidly firing neurons in the dorsal raphe of the rat was examined using extracellular recording and intracellular injection of horseradish peroxidase. Rapidly firing neurons (termed F-cells in this report) continue to fire at high spontaneous rates during intracellular recording. This and their brief (ca. 1 ms) and symmetrical action potentials distinguish them from the slowly firing, presumably serotonergic neurons in dorsal raphe. Intracellular labeling with horseradish peroxidase reveals that F-cells have small (10-15 microns) spherical, multipolar or piriform somata. Somatic spines are sparse or entirely absent. The general form of the dendritic tree is radiant and poorly branching. However, the dendrites of F-cells take two forms, with both forms being present on the same neuron so that besides having a complement of poorly branching dendrites, each F-cell has at least one dendrite with a slightly tufted branching pattern: a short primary dendrite gives rise to 3 or 4 secondary branches. The axons of F-cells project from the nucleus. They align themselves along the trajectories of known dorsal raphe efferent pathways, coursing laterally and ventrorostrally beyond the bounds of the nucleus. Morphometric measurements of retrogradely labeled dorsal raphe projection neurons provide additional evidence that small projection neurons exist. PMID- 3030493 TI - Angiotensin II sensitive neurons in the supraoptic nucleus, subfornical organ and anteroventral third ventricle of rats in vitro. AB - The angiotensin II (AII) sensitivity of neurons in the supraoptic nucleus (SON), subfornical organ (SFO) and the region near the anteroventral part of the third ventricle (AV3V) was investigated using extracellular recording in the rat brain slice preparation by adding AII (10(-10)-10(-6) M) to the perfusion medium. Forty seven (44%) of 106 SON neurons, 62 (66%) of 94 SFO neurons and 28 (33%) of 86 AV3V neurons were excited by AII. One cell was inhibited by AII in the SON and one in the SFO. The threshold concentration to evoke responses in the SON neurons was approximately 10(-9) M, but neurons in the SFO and AV3V showed clear excitatory responses to AII at 10(-10) M. In the SON, 18 (40%) of 45 phasic firing neurons (putative vasopressin neurons) and 29 (48%) of 61 nonphasic firing neurons (including putative oxytocin neurons) were excited by AII. The excitatory effect of AII was reversibly antagonized by a specific antagonist saralasin and persisted after synaptic blockade in medium with low [Ca2+] and high [Mg2+]. We conclude that AII can stimulate both vasopressin and oxytocin release, acting directly upon SON neurons and also that both the SFO and AV3V are important receptive sites for AII (although the SFO is relatively more sensitive) which contributes SON input and modulates release of these hormones. PMID- 3030494 TI - The pattern of [3H]cyclofoxy retention in rat brain after in vivo injection corresponds to the in vitro opiate receptor distribution. AB - Cyclofoxy, a fluorinated analog of naltrexone, has been designed specifically to permit in vivo labeling of opiate receptors in experimental animals and ultimately, humans. Recently, using positron emission tomography (PET), 3 [18F]acetylcyclofoxy was shown to accumulate in opiate receptor-rich brain regions of a live baboon and to be stereospecifically displaced by injections of (-)-naloxone but not (+)-naloxone. Autoradiographic evidence is presented here that the unacetylated compound, [3H]cyclofoxy, labels a population of opiate receptors in brain after in vivo injections that is virtually identical to that labeled by [3H]naloxone. The in vivo binding patterns of [3H]cyclofoxy in brain were similar to those obtained following incubation of slide-mounted brain sections in vitro. Intravenous injections of [3H]cyclofoxy to rats yielded high (greater than 4:1) striatal and thalamic to cerebellar binding ratios in brain homogenates, supernatants and in 24 micron-thick brain sections 60 min after 30 mu Ci per animal (spec. act. = 16.4 Ci/mmol). Autoradiographs revealed the typical opiate antagonist binding profile with marked retention of [3H]cyclofoxy in the striatal patches, subcallosal streak, medial habenula and central thalamus with little retention of label in cerebellum. [3H]Cyclofoxy binding was reversible since the radiolabeled drug disappeared from brain tissue within 2 h after injections to rats, or could be removed from brain slices in vitro by washing slide-mounted tissue sections for 45 min. In addition, after in vitro washing the same brain sections again bound [3H]cyclofoxy or [3H]naloxone in the same pattern. When pre-washed brain sections were incubated with [3H]cyclofoxy in the presence of unlabeled naloxone, [3H]cyclofoxy binding was reduced to background levels. These data show that [3H]cyclofoxy labels-sensitive opiate receptors in vivo and in vitro. The present results combined with evidence that cyclofoxy demonstrates a low level of toxicity in animals suggest that cyclofoxy is an excellent tool with which to study the physiological role of opiate receptors in living animals using in vivo autoradiography, and in humans using PET. PMID- 3030495 TI - Mechanisms of halothane action on synaptic transmission in motoneurons of the newborn rat spinal cord in vitro. AB - Action of halothane on synaptic transmission was studied on the isolated newborn rat spinal cord. Clinical doses of halothane (less than or equal to 3%) suppressed mono- and polysynaptic reflexes, dorsal root reflexes, excitatory and inhibitory postsynaptic potentials as well as the spontaneous synaptic potentials caused by impulse bombardment. However, the spontaneous miniature inhibitory postsynaptic potentials observed after blocking impulse activities by tetrodotoxin were not all suppressed by halothane. During halothane administration, the membrane potential of motoneurons was hyperpolarized by several millivolts, associated with an increase in input conductance. However, the threshold potential level for spike generation was virtually unaffected. Depression of synaptic transmission in spinal motoneurons by halothane is suggested to be due to two factors: a reduction in the amount of transmitter release secondary to interference with Ca2+ entry into nerve terminals, either by partial blockade of impulse invasion or voltage-dependent Ca2+ channels; and an increase in the depolarizing current necessary for excitation of motoneurons owing to hyperpolarization and decreased input resistance. PMID- 3030496 TI - REM sleep deprivation up-regulates adenosine A1 receptors. AB - Adenosine receptor binding was determined in the brains of rats deprived of rapid eye movement (REM) sleep for 48 and 96 h using [3H]L-phenylisopropyladenosine. Adenosine A1 receptors (Bmax) were significantly increased in the cortex and corpus striatum, and this increase was sleep-specific. Endogenous adenosine was assayed in microwave-fixed brain tissue and no significant changes were found in REM-deprived rats. PMID- 3030497 TI - The corticostriatal projection in cat: relation between axon terminals and evoked potentials. AB - Because of the potential value of evoked potential methods in evaluating striatal organization, a study was made to examine the reliability of evoked potentials in demonstrating the fine topographic details of the corticostriatal projection in comparison with the autoradiographic fiber-tracing method. The present data indicate a close relation between the distribution of striatal evoked potentials to electrical stimulation of a specific site in the motor cortex and the distribution of axon terminals emanating from that same cortical site in the same animal. PMID- 3030498 TI - Application of spin-trapping study to rat ischemic brain homogenate incubated with NADPH and Fe-EDTA. AB - A spin-trapping technique was applied to the detection of free radicals generated in nicotinamide-adenine denucleotide phosphate (NADPH)-dependent lipid peroxidation of rat ischemic brain homogenate. The spin adduct of phenyl-t butylnitrone (AN = 16.2-16.5 g, AH beta = 3.6-3.8 g) was observed, and it was thought to be derived from NADPH-dependent lipid peroxidation from its oxygen and NADPH dependency. Its intensity was increased in the recirculated condition following ischemic insult, indicating the susceptibility of brain tissue to lipid peroxidation in such a condition. PMID- 3030499 TI - Neurotransmission parameters estimated from miniature endplate current growth phase. AB - A numerical model of miniature endplate current (mepc) generation was fitted to the rising phase of individual mepcs recorded at the frog neuromuscular junction, and estimates of 6 transmission parameters were obtained. Model fitting was enabled by assuming literature values for geometric parameters and determining single channel current by noise analysis, the channel closing rate constant from the mepc decay, and acetylcholine hydrolysis parameters from mepcs recorded in esterase-blocked endplates. Under control conditions, mean estimates were: number of molecules in a quantum = 29,000, diffusion coefficient = 2.8 X 10(-6) cm2s-1, endplate receptor density = 8500 micron-2, forward binding rate constant = 7.6 X 10(8) M-1s-1, equilibrium dissociation constant = 58 microM and channel opening rate constant = 8100 s-1. PMID- 3030500 TI - The population of the dorsal root ganglion cells which have central processes in ventral root and their immunoreactivity. AB - Axonal transport of fluorescent dyes applied to the cut distal ends of rat L4 dorsal and ventral spinal roots was studied in order to characterize the population of dorsal root ganglion (DRG) neurons emitting axons entering the ventral root. Ca. 9% of DRG neurons, principally of small or medium size, can be labeled from the ventral root, and 55% of these display immunoreactivity for the most ubiquitous DRG neuropeptide, calcitonin gene-related peptide (CGRP). Attempts to simultaneously label cut dorsal and ventral roots revealed that double labeling was exceedingly rare and that dorsal root labeling was markedly reduced. The results are consistent with previous reports of small DRG cells emitting axons which loop into the ventral root before entering the spinal cord via the dorsal root. The few cells labeled simultaneously from cut dorsal and ventral roots indicate that axonal bifurcation distal to the DRG is very rare. PMID- 3030501 TI - Aluminum alters cyclic AMP and cyclic GMP levels but not presynaptic cholinergic markers in rat brain in vivo. AB - Oral administration of 0.3% aluminum (citrate or sulfate salt) for 4 weeks significantly elevated adenosine 3',5'-monophosphate (cyclic AMP) levels in rat cortex, hippocampus, striatum and cerebellum. The largest effect observed was a 60% increase in cortical cyclic AMP levels in rats administered aluminum sulfate. The effects of orally administered aluminum on guanosine 3',5'-monophosphate (cyclic GMP) levels were less widespread. Dietary aluminum citrate only elevated cyclic GMP levels in the hippocampus, while aluminum sulfate caused significant increases in the cerebellum, hippocampus and striatum. Aluminum citrate administered i.c.v. (1 mumol, 2 weeks postadministration) elevated cyclic AMP levels in the cortex, but had no effect on cyclic GMP levels. Aluminum administered either orally or i.c.v. had no effect on in vivo acetylcholine levels. However, dietary aluminum citrate significantly reduced choline levels in the cortex, hippocampus and striatum. Aluminum administered i.c.v. had no effect on choline acetyltransferase activity or on high-affinity choline transport. These results indicate that: the metabolism of cyclic AMP and of cyclic GMP are more sensitive to aluminum than are presynaptic cholinergic processes; the metabolism of cyclic AMP is more sensitive to the effects of aluminum than is the metabolism of cyclic GMP; and cortical cAMP metabolism is the most sensitive to the presence of aluminum. Possible consequences of elevated levels of cyclic nucleotides induced by aluminum in the brain are proposed. PMID- 3030502 TI - Protection from ischemia-induced cerebral edema in the rat by U-50488H, a kappa opioid receptor agonist. AB - U-50488 is a specific kappa opioid agonist which produces in rats water diuresis resulting in an elevation of plasma osmolarity. Pretreatment with U-50488H (the methanesulfonate salt) in Fisher rats prior to 4 h of bilateral carotid occlusion prevented the development of edema in the forebrain, and the effect was greater than that from pentobarbital anesthesia. An additional injection of an antidiuretic hormone which prevented the plasma hyperosmolarity also significantly reduced the anticerebral edemic effects of U-50488H. The plasma osmotic effect, however, may not completely account for the ischemic protection produced by U-50488H. PMID- 3030503 TI - The effect of hypophysectomy, ACTH fragments and thalamic lesions upon kindled epilepsy. AB - Hypophysectomized rats showed aberrant and retarded rates of kindled epilepsy. In hypophysectomized rats administered adrenocorticotropic hormone (ACTH) subunits ACTH4-10 and ACTH1-24, the normal kindling pattern was restored. However, in hypophysectomized animals which also had lesions of the thalamus (nucleus parafascicularis), ACTH4-10 did not restore the normal pattern of kindling. There have been many conjectures that kindling may be a subcase of learning. These results are compatible with this hypothesis, since the same procedures act in an analogous fashion within avoidance conditioning paradigms. PMID- 3030504 TI - Development and plasticity in hamster trigeminal primary afferent projections. AB - At birth (gestational day 16), the hamster infraorbital nerve projects to the appropriate portion of the brainstem, though the projection lacks adult-like internal organization (patchiness). Infraorbital nerve damage at this time does not produce appreciable transganglionic atrophy in the central projections of the infraorbital nerve, but it does result in a failure to develop normal infraorbital primary afferent patches. Such damage also produces a more widespread central projection of spared mandibular afferents into regions occupied by 'regenerate' infraorbital terminals (J. Comp. Neurol., 235 (1985) 129 143). In the present study, transganglionic transport techniques were again used to show that, by postnatal day 5 (gestational day 21), rostrocaudally continuous aggregates of horseradish peroxidase-labelled infraorbital terminals are visible throughout the trigeminal brainstem nuclear complex. This aggregation pattern is nearly adult-like and isomorphic with the distribution of the mystacial vibrissae on the face. A similar infraorbital lesion performed on postnatal day 5, however, markedly decreased the density of the adult central projection of the infraorbital nerve to subnuclei principalis, oralis, interpolaris, and the magnocellular laminae of caudalis. The projection to superficial laminae of caudalis and the cervical dorsal horn was maintained. A postnatal-day-5 infraorbital lesion also failed to produce a more widespread central projection from spared mandibular primary afferents. These data suggest a relationship between the postnatal maturity of trigeminal primary afferents and the response of damaged and undamaged trigeminal afferents to infraorbital nerve transection in hamster. The similarity in the central primary afferent response to lesions at equivalent gestational times (postnatal days 5 and 0, respectively) in hamster and rat, suggests that this plasticity gradient may be a general characteristic of mammalian trigeminal primary afferents. PMID- 3030505 TI - Dolichol kinase and the regulation of dolichyl phosphate levels in developing brain. AB - The developmental changes of dolichol kinase activity and dolichyl phosphate levels have been studied in rat brain. Because both dolichol kinase activity and dolichyl phosphate were enriched in microsomes, detailed study of this subcellular fraction was carried out. Dolichol kinase specific activity in brain microsomes increased postnatally 3-fold to a maximum at ca. 30 days of age. This increase was observed whether exogenous dolichol was present or not and whether Zn2+ or Ca2+ was utilized as the cation for the enzyme. Zn2+ was the most effective cation in developing brain, as we have shown previously for adult brain (Sakakihara, Y. and Volpe, J.J., J. Biol. Chem., 260 (1985) 15413-15419). Although the Vmax for the enzyme increased by three-fold with development, the Km for dolichol and for CTP did not change, indicating that the developmental increase was not related to an alteration in catalytic efficiency of the enzyme. A striking and parallel increase in dolichyl phosphate levels in brain microsomes was defined with development. Levels were lowest in one-day-old animals and then increased ca. 13-fold to a maximum at 30 days of postnatal age. The parallel increase in dolichol kinase activity and dolichyl phosphate levels in microsomes of developing brain suggests that dolichol kinase is the principal determinant of cellular levels of dolichyl phosphate, the critical intermediate in the dolichol linked pathway to glycoproteins. PMID- 3030506 TI - Differential ontogeny of divalent cation effects on rat brain delta-, mu-, and kappa-opioid receptor binding. AB - The effect of the divalent cations, Mn2+, Mg2+ and Ca2+ on rat forebrain delta-, mu- and kappa-receptor binding was examined during postnatal development. It was found that delta-receptor binding, assessed with [3H]D-Ala2-D-Leu5-enkephalin ([3H]DADLE) (+ 10 nM D-Ala2- MePhe4-Gly-ol5-enkephalin (DAMGE)), was stimulated by the 3 cations in a dose- and developmental time-dependent manner. delta Binding was most sensitive to the cations during the first week postnatal, prior to the appearance of high-affinity delta-binding. In contrast, inhibition of mu receptor binding ([3H]DAMGE) by divalent cations appeared early in development and remained constant throughout the postnatal period. Divalent cation inhibition of kappa-binding ([3H]ethylketocyclazocine ([3H]EKC) + 100 nM DAMGE and 100 nM DADLE) appeared after the second week postnatal. These results demonstrate that the characteristics and postnatal development of divalent cation modulation of mu , delta- and kappa-binding is distinctly different. Thus, the neonate may be a good model system to examine the binding properties and functions of delta- and kappa-receptor subtypes. PMID- 3030507 TI - Tests for relabelling the goldfish tectum by optic fibers. AB - The temporal half of retina was removed in goldfish and the optic nerve was crushed. Electrophysiological recordings at 1-2 months revealed a retinotopic expansion of the nasal half retina across nearly the entire tectum. In other fish, an abnormal innervation of nasal fibers in anterior tectum was induced and allowed to persist for 51 months. When fibers were then crushed, nasal fibers innervated posterior tectum at 1-2 months showing no preference for anterior tectum. Apparently, fibers did not relabel tectum. PMID- 3030508 TI - Norepinephrine regulates long-term potentiation of both the population spike and dendritic EPSP in hippocampal dentate gyrus. AB - Hippocampal slices from norepinephrine (NE)-depleted rats exhibited marked reductions in long-term potentiation (LTP) of both the population spike and dendritic EPSP in the dentate gyrus. In contrast, depletion of serotonin (5 hydroxytryptamine, 5-HT) had no effect on either population spike or EPSP-LTP. In addition, superfusion of slices with NE produced potentiation of both the granule cell population spike and dendritic EPSP which persisted long after NE washout. These data support a role for NE in regulating long-term plasticity of both granule cell action potential firing and dendritic EPSPs. PMID- 3030509 TI - "Peripheral" benzodiazepine binding sites in the Maudsley reactive rat: selective decrease confined to peripheral tissues. AB - "Peripheral" benzodiazepine binding sites (PBS) were studied in both the CNS and peripheral tissues of Maudsley reactive (MR) and Maudsley non-reactive (MNR) rats using the PBS specific ligand [3H]Ro 5-4864. A statistically significant reduction in the density of PBS was found in heart and kidney of the MR compared to the MNR. Similar reductions in the density of PBS were not observed in a number of areas of the central nervous system (including cortex, hippocampus, and hypothalamus) or other peripheral tissues, such as lung and adrenal. This selective decrease in PBS in a strain selectively bred for a high degree of "fearfulness" may be related to previous findings of a reduction in the density of PBS in the same tissues in rats subjected to uncontrollable shock. These observations suggest that PBS in heart and kidney may be altered in response to fear or anxiety. PMID- 3030510 TI - Theta in hippocampal slices: relation to synaptic responses of dentate neurons. AB - The effects of cholinergic agents on isolated dentate neurons were studied to characterize cellular mechanisms underlying carbachol-induced 'theta' EEG activity. Carbachol, eserine, and acetylcholine produced a synchronization of slow wave activity (theta) accompanied by depression of perforant path to dentate field potentials. These effects were antagonized by atropine but not d tubocurarine. The results suggest that muscarinic receptors mediate theta activity resulting from a depolarization of dentate neurons. PMID- 3030511 TI - Synaptic connections of thalamo-cerebral vocal nuclei of the canary. AB - The canary vocal nuclei include two systems: hyperstriatum ventrale, pars caudale (HVc), nucleus robustus archistriatalis (RA) and nucleus hypoglossus, pars tracheosyringealis (nXIIts) compose a motor driving system for vocalization. The other group of nuclei including HVc, nucleus X of the lobus parolfactorius (Area X), nucleus dorsointermedius posterior thalami (DIP) and nucleus magnocellularis of anterior neostriatum (MAN) is a modulator of the driving system. The HVc neurons receive mono- or polysynaptic innervation from the MAN and send their fibers to Area X. Axons of Area X terminated in the thalamic nucleus, the DIP, from which neurons extended axons back to the cerebral nucleus, the MAN. Accordingly, the HVc, Area X, DIP and MAN are in a closed loop. Auditory inflow may converge on HVc neurons, partly from either the MAN or Area X during feedback control of the song. Thalamic neurons in the DIP responded to MAN stimulation, and to tonal stimuli, with relatively long latency. The interconnections between the HVc and MAN neurons are presumably central in voco-auditory integration during song-learning. PMID- 3030513 TI - [Enzootic bovine leukosis and feline leukosis. 2 current animal models for the study of virus-induced lymphomas and immunodepression syndrome in man]. PMID- 3030512 TI - Labelling of peripheral-type benzodiazepine binding sites in human brain with [3H]PK 11195: anatomical and subcellular distribution. AB - The peripheral-type benzodiazepine binding site, erstwhile characterized in the rodent and feline brain, has now been characterized in post-mortem human brain using [3H]PK 11195. The kinetics and pharmacological properties of the binding of this ligand are similar to peripheral-type benzodiazepine binding sites elsewhere. The potency of RO5-4864 for this site in human brain is close to that seen in ruminant and carnivore tissues but considerably lower than in rodent tissues. The regional distribution of these binding sites would suggest a neuronal rather than a glial localization. [3H]PK 11195 bound in a similar fashion to slide-mounted sections of human brain, thus allowing quantitative studies of the regional distribution of peripheral-type benzodiazepine binding sites to be made. The binding sites were distributed heterogeneously, but were restricted to the grey matter. Highest densities of binding sites were found in forebrain structures. The localization was not limited to any functional system, nor did it resemble any previously described transmitter system. The similarities between peripheral-type benzodiazepine binding sites in human and in feline brain in terms of their pharmacological characteristics and their regional and subcellular distribution suggest that the cat, rather than the rat, may be the better model for studying a possible role for this site in human cerebral function. PMID- 3030514 TI - Brain surface exposure to CO2 and NdYag laser application: effect on pulmonary endothelium response in the rat. AB - Pulmonary endothelial cells and the renin-angiotensin-aldosterone (RAA) system respond to different types of injury (direct or indirect) with variations in their functions. These variations influence the regulatory mechanisms of pulmonary and systemic blood pressure, electrolyte balance and fibrinolysis. Concentration changes of some components of the RAA system and lung plasminogen activator were observed following NdYag laser application to the brain surface in rats. These changes were similar to those observed in cutaneous burn and haemorrhagic hypotension. CO2 laser application did not cause the same changes. PMID- 3030515 TI - Ectopic hormone syndromes. PMID- 3030516 TI - [High correlation between isoenzyme classification and kinetoplast DNA variability in Trypanosoma cruzi]. AB - By means of 14 restriction enzymes, we have studied the kinetoplast DNA polymorphism in 21 Trypanosoma cruzi isolates previously classified into 19 different genotypes based on the analysis of 15 isozyme loci. We have found a high correlation (p less than 0.001) between the proportion of restriction bands (fragments) common to any two given isolates and the corresponding genetic identities calculated from the isozyme data. This shows that the two classifications (kDNA and isozymes) corrobate one another and strongly suggests that the two types of variability are correlated with time (molecular clocks). The phylogenic classifications so obtained can be used as rational bases for medical and epidemiological studies. Although they are correlated, the two types of analysis are complementary as they do not yield identical results. Like the isozyme genetic distances and genetic identities, the values obtained for the proportion of common restriction bands (fragments) exhibit a continuum. This seems to confirm that natural T. cruzi populations exhibit a wide range of genotypes rather than a few well-differentiated clusters of strains. PMID- 3030517 TI - [Biochemical manifestations of the poisoning of larvae of Aedes aegypti (Insecta, Diptera) by the delta-endotoxin of Bacillus thuringiensis israelensis. I. Hemolymph carbohydrates]. AB - The glycemia of 4th instar larvae of Aedes aegypti (Diptera Culicidae) was determined over a 30 min to 36 hrs period following their immersion in water containing 0.00-0.01-0.02-0.1-1.0 mg per liter of a 12,000 I. U. per mg preparation of the delta-endotoxin of Bacillus thuringiensis israelensis. Seven major carbohydrate fractions were separated in the haemolymph, among which glycogen, trehalose and glucose were identified; the most concentrated of the 4 remaining unknown fractions, exhibits a similar chromatographic behavior but different chemical properties than glucuronic acid. The glycogen fraction shows two large amplitude peaks at 1 hr 30 and 6 hrs with 0.1 mg/l, whereas with the 2 lower doses (0.01 and 0.02 mg/l) a first drop from 30 min to 1 hr is followed by a peaking at 12 hrs. The haemolymph trehalose concentrations of intoxicated larvae increase relatively to controls 2 hrs after incubation with the lower dose; by contrast, they decrease with the 2 higher doses. The glucosemia increases in all cases, but more markedly with the higher dose as soon as 1 hr following the treatment. A positive correlation was found over the whole treatment duration between the glucosemia nd the trehalosemia of control larvae. The regression slope of this relationship decreases with increasing toxin doses and turns to negative values with 0.1 and 1.0 mg/l. These observations suggest the hypothesis that a uncontrolled increase of trehalase activity develops according to the increasing stages of intoxication. This clearly differenciates the effects of the B. T. i toxin from those of parasites, such as Fungi, Sporozoans or Nematodes in Mosquitoes. PMID- 3030518 TI - Osteoblastlike cells in a serum-free methylcellulose medium form colonies: effects of insulin and insulinlike growth factor I. AB - Osteoblastlike (OB) cells obtained from a heterogeneous primary cell population by enzymatic cell digestion of calvaria of newborn rats are grown in a serum-free viscous alpha-MEM/F-12 medium containing 0.8% methylcellulose. In contrast to cell monolayers in conventional tissue cultures OB cells proliferate into colonies of rounded-up cells to form morulalike spherical cell clusters containing up to 100 cells. These colonies, with different cell numbers, are clearly not fibroblastlike since fibroblasts from the same rats always grow as a cell monolayer. Alkaline phosphatase activity and cAMP responsivness to PTH are expressed more markedly (70% and 250% respectively) by OB cells in the described culture system than in conventional tissue cultures. Rounded-up OB cells sediment and colonies stick to the dish; proliferation of OB cells is favored and starts 3 4 days after inoculation. Increasing concentrations of insulinlike growth factor (IGF) I (0.4-35 nM) and insulin (20-660 nM), as well as increasing initial cell density, enhances mitogenic activity of these cells in a dose-dependent way. On a molar ratio IGF I (physiological concentrations) is 10 times as potent as insulin (pharmacological concentrations) with respect to proliferation. If less than 10(5) cells/ml are inoculated, there exists an apparent relationship between initial cell density and major onset of replication, indicating the presence and accumulation of local growth factors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3030520 TI - The effect of nifedipine on ischemia-induced changes in the biochemical properties of isolated sarcolemmal vesicles and the ultrastructure of myocardium. AB - To investigate whether slow Ca2+ channel blockers protect against development of changes in properties of the sarcolemma and in the tissue ultrastructure during myocardial ischemia, nifedipine was administered prior to occlusion (up to 3 hours) of the left anterior descending coronary artery in anesthetized pigs. Intravenous doses which reduced arterial blood pressure by 20-25%, had no effect on the time-dependent reduction of Ca2+-calmodulin and cyclic AMP-dependent 32P incorporation into sarcolemmal phospholamban-like protein. Nifedipine blocked the reduction in the activity of sarcolemmal 5'-nucleotidase. Nifedipine had no significant effect on the long-chain fatty acylcarnitine accumulation in sarcolemma. A marked delay in the appearance of ultrastructural indicators of irreversible tissue injury in subepicardial myocardium was observed, when nifedipine was infused. Particularly the reduced appearance of electron-dense bodies in mitochondria suggested a reducing effect of nifedipine on cellular net gain of Ca2+. Apparently, ischemia-induced loss of the ability of the proteinkinases to incorporate phosphate into sarcolemmal phospholamban-like protein is not a process secondary to Ca2+ overload of the myocardium. The involvement of accumulation of long-chain fatty acylcarnitine within the sarcolemma may also be excluded. The membrane defect as indicated by a change in phosphorylation-mediated control of Ca2+ transport may itself be associated with the development of ischemia (-reperfusion)-induced Ca2+ overload. PMID- 3030519 TI - Influence of cell isolation and incubation procedures on Ca2+ dependence of glucose transport in isolated cardiac myocytes. AB - Insulin stimulates hexose transport in isolated myocytes of adult rats but data are conflicting regarding the Ca2+-dependence of this effect, demonstrated previously in intact heart muscle. 3-O-methyl-D-glucose and 2-deoxy-D-glucose transport was compared in cells prepared and incubated by different published procedures. Ca2+-dependence of hexose transport stimulation by insulin was found only with incubation in a modified Joklik tissue culture medium buff red with bicarbonate at pH 7.4. At pH 7.0 or when bicarbonate was replaced by N-2 hydroxyethylpiperazine-N'-2 ethane sulfonic acid (HEPES) stimulation by insulin was not Ca2+-dependent, nor was there a Ca2+-dependent increment in insulin effect in physiological saline media regardless of pH or buffer (HEPES, 3-[N morpholino] propanesulfonic acid (MOPS) or bicarbonate). The ATP content of cells incubated in HEPES or MOPS media was reduced. Ca2+-dependence seems to require the presence of bicarbonate and another factor(s) in tissue culture medium. The response is also influenced by the isolation procedure and may be related in part to preservation of an intact glycocalix. Thus, details in cell isolation and incubation procedures may strongly influence the behaviour of the cells. Isolated cardiac myocytes are acceptable as a valid model only if it can be demonstrated in each specific case that the characteristics present in intact tissue are maintained unaltered in the isolated cells. PMID- 3030521 TI - Murine plasmacytomas constitute a class of natural heat-shock variants in which the major inducible hsp-68 gene is not expressed. AB - Analysis of the heat-shock response in murine plasmacytomas reveals that, as demonstrated previously for the MPC-11 cell line, the genes coding for the 68 kilodalton heat-shock protein (hsp-68) are not expressed upon heat shock or sodium arsenite treatment. Noninduction is unique to the normally coordinated set of three hsp-68 genes since at least two other heat-shock protein genes (hsp-70 and hsp-89) are properly induced. No other lymphoid cell line was found to possess silent hsp-68 genes. Cell lines examined included a T lymphoma, a pre-B lymphocyte, and a non-B-non-T tumor cell line, as well as an Ig-nonproducing myeloma of undetermined differentiated status. Nonexpression is strain independent as observed in BALB/c and C3H plasmacytomas. Based on S1 nuclease analysis using a cloned genomic hsp-68 probe, nonexpression is caused by the absence of hsp-68 mRNA following heat shock. A time-course experiment suggests that rapid degradation of mRNA does not occur, implying that the block is most likely at the transcriptional level. Southern blot analysis does not indicate any minor deletions around the region of transcription initiation, at least in the probed hsp-68 gene. These results suggest that the absence of hsp-68 gene expression may be a reflection of the differentiated and (or) transformed state of murine plasma cells, possibly through the absence or deregulation of a regulatory factor required for induction of heat-shock genes. PMID- 3030523 TI - Adult ventricular myocytes isolated from CHF 146 and CHF 147 cardiomyopathic hamsters. AB - Single ventricular myocytes have been isolated from normal and cardiomyopathic hamsters using a collagenase method. The procedure produces calcium-tolerant myocytes that are viable and appear on light and electron microscopic examination to be healthy. These cells respond to electrical stimulus and have normal intracellular resting and action potentials. PMID- 3030522 TI - Nonstereoselective aspects of propranolol pharmacodynamics. AB - Twenty years after its discovery, the beta-adrenergic blocking agent propranolol continues to interest pharmacologists and clinicians. Its therapeutic profile has extended to areas beyond the purview of the cardiovascular system, and its ocular and central nervous system effects have been well documented. In addition, it still remains a very good pharmacological tool to map out the adrenergic beta receptors in the body, and stereoisomers of propranolol and other beta-blockers serve as valuable agents to distinguish between the effects related to beta adrenoceptors and those which are not. The primary purpose of this review is to summarize the evidence indicating that beta-adrenergic blocking agents lack stereoselectivity in some of their effects, including several of considerable therapeutic importance. Because many pharmacological actions of propranolol followed a nonsteroselective pattern, the involvement of beta-adrenoceptors in them was questioned and this led to the search for alternate mechanisms to explain these effects. Studies with propranolol and some related drugs indicated the involvement of a cholinergic mechanism in their antiarrhythmic, ocular hypotensive and some central effects. Also, a presynaptic inhibitory effect at the skeletal neuromuscular junction has been suggested to explain the benefical effect of propranolol and other beta-blockers in tremor. Biochemical studies with these drugs revealed their inhibitory action on the cholinesterase enzyme in blood and other tissues like myocardium and brain. It is thus hypothesized that modulation of cholinergic neurotransmission by propranolol could explain some of its nonstereoselective actions and open new vistas in propranolol pharmacodynamics. PMID- 3030525 TI - Pancreatic islet-cell neoplasia, with secretion of a parathormone-like substance and hypercalcemia. AB - In a 47-year-old woman with a pancreatic mass associated with hypercalcemia and mental confusion, medical measures failed to restore her serum calcium level to normal. To do so, radical resection of a locally invasive vascular neoplasm arising from the body and tail of the pancreas was necessary. The neoplasm was a pancreatic islet-cell tumour. Serum parathormone assays demonstrated abnormally high secretion of a parathormone-like substance. Ectopic secretion of such substances from islet-cell tumours should be considered in association with refractory metabolic disturbances. In view of the often indolent clinical course of islet-cell tumours and the potential for life-threatening hormonal effects, biopsy confirmation of adenocarcinoma should be obtained before resorting to palliative surgical management of pancreatic neoplasms. PMID- 3030524 TI - L-648,051, sodium 4-[3-(4-acetyl-3-hydroxy-2-propylphenoxy)- propylsulfonyl] gamma-oxo-benzenebutanoate: a leukotriene D4 receptor antagonist. AB - L-648,051, sodium 4-[3-(4-acetyl-3-hydroxy-2-propylphenoxy) propylsulfonyl]-gamma oxo-benzenebutanoate is a selective and competitive inhibitor of [3H]leukotriene D4 (KB value of 4.0 microM) and to a lesser extent [3H]leukotriene C4 (Ki value of 36.7 microM) binding in guinea pig lung homogenates. Functionally, it selectively antagonized contractions of guinea pig trachea induced by leukotrienes C4, D4, E4, and F4 in concentrations that did not antagonize contractions induced by acetylcholine, histamine, serotonin, prostaglandin F2 alpha, or U-44069 (endoperoxide analogue). Schild plot analysis indicated that L 648,051 competitively antagonized contractions of guinea pig ileum induced by leukotriene D4 (pA2 7.7) and contractions of trachea induced by leukotrienes D4, E4, and F4 (pA2 7.3, 7.4, and 7.5, respectively). Contractions of guinea pig trachea induced by leukotriene C4 were inhibited in a noncompetitive fashion (Schild plot slope, 0.45). Developed contractions of trachea induced by the leukotrienes were rapidly reversed by L-648,051 greater than FPL-55712 greater than L-649,923. Intravenous L-648,051 selectively blocked bronchoconstriction induced in anaesthetized guinea pigs by intravenous leukotrienes C4, D4, and E4 but not that induced by arachidonic acid, serotonin, U-44069, or acetylcholine. The compound displayed poor activity following intraduodenal administration. The profile of activity for L-648,051 indicates that it may be a useful topical agent for studying the role of leukotrienes in diseases such as bronchial asthma. PMID- 3030526 TI - Papillomaviruses in human cancer. PMID- 3030527 TI - Type 1a glycogen storage disease with hepatoblastoma in siblings. AB - Clinical and histologic details of the two siblings with type 1a glycogen storage disease (GSD-1a) who developed hepatoblastoma are presented. The light microscopic studies on hepatic tumor in both siblings revealed fetal type of hepatoblastoma. Ultrastructural findings in Patient 2 showed markedly altered mitochondria, which were frequently surrounded by the rough endoplasmic reticulum. This is the first known occurrence with this association, and the third report on the familial occurrence of this neoplasm. Glycogen storage disease may increase the risk of hepatocellular carcinoma and hepatoblastoma. PMID- 3030528 TI - Small cell carcinoma of the prostate. Part I. A clinicopathologic study of 20 cases. AB - Clinical information and histological slides of 20 cases of small cell carcinoma of the prostate seen at The University of Texas M. D. Anderson Hospital and Tumor Institute at Houston over a 23-year period were reviewed. Patient's ages ranged from 30 to 89 years (median, 67 years). In nine cases, pure adenocarcinoma of the prostate preceded recognition of the small cell component by 7 months to 8 years (median, 18 months); five of these were initially at Stage A. There was a small cell component at presentation in 11 cases (10, Stage D). Small cell carcinoma was merging with the adenocarcinoma in 11 cases and represented 30% to 90% of total tumor volume. Eleven of 20 patients died of their disease. Those presenting initially with a pure adenocarcinoma survived between 7 months and 9 years (median, 24 months). After the recognition of the small cell carcinoma component, regardless of a prior history of adenocarcinoma, death followed within 1.5 years (median, 5 months). This study suggests a biologic difference in behavior in prostatic carcinoma containing a small cell carcinoma component. The small cell component may manifest early or late in the disease. PMID- 3030529 TI - Axillary lymph node dissection for intraductal breast carcinoma--is it indicated? AB - One hundred patients with intraductal breast carcinoma (DCIS) were treated with either mastectomy (49 patients) or radiation therapy (51 patients). All patients underwent axillary lymph node dissection (average number of nodes removed, 16) as part of their treatment. No patient had any positive axillary lymph nodes. There has been one recurrence in each treatment group (median follow-up, 27 months) and no deaths. Intraductal breast carcinoma has little potential for metastasis to axillary lymph nodes. PMID- 3030530 TI - Anticancer effects of arterial administration of the anticancer agent SMANCS with lipiodol on metastatic lymph nodes. AB - A new method of arterially administering an oily anticancer agent was successfully established for the selective targeting of metastatic lymph nodes. A high molecular weight anticancer agent, a conjugate of copolymer (styrene maleic acid) to neocarzinostatin (SMANCS) was prepared in our laboratory and dissolved in a lymphographic oily contrast medium, Lipiodol (SMANCS/Lipiodol). SMANCS/Lipiodol was administered intraoperatively to eight patients with colorectal cancer and preoperatively to one patient with gastric cancer with lymph node metastases. In six of the patients with colorectal cancer, the drug was administered via an artery and in the other two patients the drug was injected into the wall of the colon near the primary cancer. In the patient with gastric cancer, the drug was administered via the left gastric artery. Delivery of the drug to the lymph nodes was examined roentgenologically and the anticancer effect was examined histologically. The results showed that SMANCS/Lipiodol could be delivered to the metastatic lymph node via the artery, but it could not be delivered to the metastatic lesion of the lymph node via the lymphatic route. In the patient with gastric cancer, SMANCS/Lipiodol preoperatively administered via an artery was found to remain selectively in a metastatic lymph node and an anticancer effect was histologically proved in all three of the metastatic lymph nodes. PMID- 3030531 TI - Patient age, histologic features, and length of survival in patients with glioblastoma multiforme. AB - Histologic sections from 71 patients with glioblastoma multiforme were reviewed to identify histologic prognostic factors and to explain the significantly shorter survival in older patients. Slides were studied for 14 histologic variables from a group of 35 patients aged less 45 years and from 36 patients aged 65 years or more. The relation of these histologic factors to the length of survival and age group was then investigated. The results document the marked histologic and cytologic heterogeneity of the glioblastoma and reaffirm the importance of necrosis as a prognostic factor. The results further suggest that patients whose glioblastomas contained microcysts, pseudopalisading, cells with astrocytic differentiation, and large areas of better differentiated glioma, did better than those patients whose lesions were homogeneously composed of small cells or whose lesion had a small median nuclear size. The study reaffirmed the strong (P less than 0.0001) negative relationship between advancing age and duration of postoperative survival. The presence of necrosis, a smaller standard deviation of nuclear size, the extent of vascular proliferation, the absence of well differentiated neoplastic fibrillary astrocytes, and neoplasms composed homogeneously of small cells were related to patient age and offered a possible explanation for at least part of the observed age effect. However, the strong relation between age and survival remained significant when adjusted for other variables, and the effect of age must rest largely on factors other than those detected in this morphologic study. PMID- 3030532 TI - Correlation between cytogenetic and molecular findings in human chronic myelogenous leukemia lines EM-2 and EM-3. AB - Few established cell lines derived from patients with chronic myelogenous leukemia have been reported. Cytogenetic examinations of two independently derived Philadelphia chromosome (Ph)-positive cell lines from a patient with chronic myelogenous leukemia were performed serially from their initiation in 1982 to the present. Subcultures of each of these lines maintained separately in two laboratories for over 2 years were compared for degree of divergence. The modal chromosome number declined substantially within the first few months and slowly thereafter. Despite hyperploidy, these lines have remained remarkably stable cytogenetically. For each line, the modal chromosome number is hypotetraploid, multiple copies of Ph are present and no normal chromosome #9 remains. Only a few marker chromosomes have arisen. Despite the multiple copies of Ph, a single bcr restriction pattern was seen, suggesting duplication of a single Ph, rather than independent translocation events. These lines should be very useful for in vitro studies of chronic myelogenous leukemia. PMID- 3030534 TI - Transposition of breakpoint cluster region (3' bcr) in CML cells with variant Philadelphia translocations. AB - A probe derived from the 3' end of the CML breakpoint cluster region (bcr) was localized in chronic myelocytic leukemia (CML) cases with complex Philadelphia translocations, [t(8;9;22)(q13;q34;q11) and t(12;9;22)(p11;q34;q11)], and with "masked" Ph chromosomes, [t(9;5;22)(q34;q31;q11) and t(9;22)(q22;q34)], by a chromosomal in situ hybridization technique. In some cases, the 3' bcr rearrangements in the DNA were examined with Southern blot analysis. In each case, a significant accumulation of grains hybridized to the 3' bcr probe was observed at chromosomal segments derived from the long arm of a chromosome #22. In some cases, the accumulation of grains was detected on both translocated segments derived from the 22q and terminal portions of Ph chromosomes; Southern blot analysis revealed that breakage in chromosome #22 occurred within DNA sequences of the 3' bcr probe involved in the Ph translocations. In a CML cell line, K562, no accumulation of grains hybridized to the 3' bcr probe was detected, except at 22q11 of the normal chromosomes #22; Southern blot analysis of this cell line revealed that the 3' bcr sequences were missing. Thus, the data presented here suggest a complexity in the formation of "masked" Ph chromosomes. PMID- 3030533 TI - Human-fms gene is retained in acute lymphoblastic leukemia cells with del(5)(q32). AB - Cytogenetic and molecular investigations of NALM 6 cells (a pre-B-lymphoblastic acute leukemia cell line) revealed them to contain both alleles of the c-fms gene, though the cells had chromosomal changes of 5q- and 12p+. The amount of DNA fragments hybridized to the 1.4 kb PstI/PstI v-fms probe in the NALM 6 cells was approximately the same, when compared with cells of an Epstein-Barr virus transformed lymphoblastoid cell line with a normal karyotype. Chromosome banding analysis revealed that the breakpoint of the 5q- in the NALM 6 cells was at the proximal portion of the 5q32 band. Chromosomal in situ hybridization of NALM 6 cells showed a significant accumulation of grains on the terminal portions of the abnormal 5q- chromosomes (5q32), as well as on the normal chromosomes #5 with a peak at 5q32-q33. These findings indicate that the human c-fms gene is not deleted in the lymphoblastic leukemia cells with a 5q- studied by us and that it does not show rearrangement or amplification. Thus, the results indicate that a difference in the dosage of the c-fms gene in acute lymphoblastic leukemia cells with the 5q- versus that in cells with the 5q- change in nonlymphocytic neoplasia; in the latter a hemizgosity of the c-fms gene has been suggested. PMID- 3030535 TI - Flow cytometric and karyotypic analysis of a primary small cell carcinoma of the prostate: a xenografted cell line. AB - A human small cell undifferentiated carcinoma of the prostate, xenografted in nude mice, was analyzed both cytogenetically and by DNA flow cytometry. The DNA content of the line indicated its stability on serial passage, and was consistent with the cytogenetic findings. The banded karyotype was hypodiploid with nonrandom losses of chromosomes #6, #7, #10, and #13. Structural rearrangements involved chromosomes #1 and #2, and there were three unidentified markers. The findings were compared with those described in other types of prostatic carcinoma. PMID- 3030536 TI - A consistent chromosome translocation in synovial sarcoma. PMID- 3030537 TI - Translocation (X;18) in a synovial sarcoma. PMID- 3030538 TI - Clofibrate does not induce peroxisomal proliferation in human hepatoma cell lines PLC/PRF/5 and SK-HEP-1. AB - The potential of fibrate drugs to induce peroxisomal proliferation in human liver cells was evaluated in athymic nude mice transplanted with human hepatoma cells and treated by clofibrate in vivo as well as in cultured human hepatoma cells in the presence of fibrate drugs added to the culture medium. Clofibrate did not induce peroxisomal activities and neither acted as a peroxisomal proliferator in human PLC/PRF/5 or SK-HEP-1 heterotransplants under conditions of induction of peroxisomal activities in the host rodent liver. Similarly, clofibric acid or bezafibrate did not induce peroxisomal activities in cultured human PLC/PRF/5 or SK-HEP-1 cells under conditions of induction of peroxisomal activities in cultured primary rat liver cells. The lack of response of the human cells to peroxisomal proliferators of the fibrate type may indicate a species specificity with respect to induction of peroxisomal activities by xenobiotic peroxisomal proliferators. PMID- 3030539 TI - Precancerous lesions of the cervix. Personal risk factors. PMID- 3030540 TI - Topoisomerase II-mediated DNA breaks and cytotoxicity in relation to cell proliferation and the cell cycle in NIH 3T3 fibroblasts and L1210 leukemia cells. AB - The DNA intercalator, 4'-(9-acridinylamino)methanesulfon-m-anisidide (m-AMSA) and the nonintercalator, etoposide (VP-16) produce topoisomerase II-mediated protein linked DNA strand breaks. This function of topoisomerase II was investigated in relation to cell proliferation and cell cycle. Mouse fibroblasts NIH 3T3 and mouse leukemia L1210 cells stop proliferation when they reach a certain density. Nuclei were isolated from proliferative or quiescent cells and then treated with drug for 30 min. DNA modifications were assayed by alkaline elution. We found that the frequencies of m-AMSA- or VP-16-induced DNA-protein links were higher in nuclei from exponentially growing than in those from quiescent cells in both the 3T3 and the L1210 lines. Drug-induced protein-associated DNA breaks were also studied as a function of the cell cycle in 3T3 cells that had been arrested by contact inhibition in medium containing 1% calf serum and then stimulated to proliferate by raplating at a lower cell density in medium containing 10% serum. In these synchronized cells, a large peak of [3H]thymidine incorporation occurred 18-30 h after replating. The yield of DNA-protein cross-links produced by 30-min drug treatments of nuclei isolated at various times after growth initiation increased concomitantly with the peak of the DNA synthesis. The topoisomerase II activity of nuclear extracts, as measured by kinetoplast DNA decatenation followed a similar pattern. Using colony-forming assays, we also observed that m AMSA and VP-16 were most cytotoxic in proliferative cells and during DNA synthesis. These results suggest that alkaline elution measurement of m-AMSA- or VP-16-induced protein-linked DNA breaks reflects the association of topoisomerase II with DNA. This association is increased during DNA replication, making the cells more vulnerable to m-AMSA and VP-16 at this time. PMID- 3030541 TI - In vitro transformation of human B-cell follicular lymphoma cells by Epstein-Barr virus. AB - Human low-grade B-cell lymphoma cells cannot be readily maintained in long-term tissue culture. In an effort to obtain low-grade B-cell lymphoma cell lines for in vitro study, we used Epstein-Barr virus (EBV) as a transforming agent. T-cells were removed prior to EBV transformation by rosetting with sheep erythrocytes, followed by treatment with anti-T11 monoclonal antibody plus complement. The resulting cell population was cocultured with EBV prepared from tissue culture supernatants of marmoset cell line B95-8. Identical immunoglobulin gene rearrangements of tumor cells and EBV-transformed cells were the criteria used to determine that the transformed cells were of tumor origin. DNA was prepared from both biopsy tissue and EBV cell lines and digested with restriction endonucleases, and Southern blots were prepared by standard methods. B-cells isolated from biopsies of four low-grade B-cell lymphomas of follicular, small cleaved cell type and one of follicular, mixed cell type were transformed by EBV into rapidly growing in vitro tissue culture lines. Two of the five transformed cell lines had immunoglobulin heavy chain and light chain gene rearrangements which were present in cells from the original tumor biopsy, indicating that these EBV-transformed cell lines are of tumor origin. PMID- 3030542 TI - Immunoelectron microscopic localization of the pX gene products in human T-cell leukemia virus type 1-producing cells. AB - The location of the pX gene products in human T-cell leukemia virus type 1 producing cells, MT-2 and HUT 102, was studied by immunoelectron microscopy using the direct and indirect peroxidase-labeled antibody methods. Fab'-peroxidase conjugates were prepared for the direct method with a maleimide compound from antisera to the carboxy-terminal region of the pX gene products. Positive immunostaining in MT-2 cells was detected in the endoplasmic reticulum, the outer and inner leaflets of the nuclear membrane, and inside their cisternae, but not in the plasma membrane and viral particles. Staining in the nucleus was faint. On the other hand, positive immunostaining in HUT 102 cells was detected diffusely in the euchromatin regions of the nucleus but not in the nucleoli, nuclear envelope, and cellular membrane systems. The location of the positive immunostaining in the HUT 102 nuclei was reconfirmed by the reaction in isolated nuclei. On the basis of both the immunoelectron microscopic and immunoblotting analyses of the pX gene products, it is suggested that the Mr 40,000 to 42,000 protein (p40x) is localized mainly in the euchromatin regions of the nuclei of human T-cell leukemia virus type 1-producing cells, and the Mr 68,000 protein (p68x) is localized mainly in the nuclear envelope and the endoplasmic reticulum of MT-2 cells. p68x detected in MT-2 cells with the anti-p40x serum was deduced to be a protein consisting of p40x and a part of env gene products and to share epitopes in common with p40x. PMID- 3030543 TI - Proliferative fibromatosis in avian skeletal muscle caused by cloned recombinant avian leukosis viruses. AB - Two molecular recombinants (EU-8 and K-3) constructed from ring-necked pheasant virus and UR2AV, the helper virus associated with avian sarcoma virus UR2, caused a high incidence of a hitherto unreported pathological condition in chick skeletal muscle. A disease spectrum was observed in which muscle was infiltrated by proliferating fibroblasts and caused white streaks, white diffuse areas, or well-defined elongated tumors. Fibroblast proliferation was progressive, and the gross presentation depended on the rapidity and extent of proliferation. There was evidence of anaplasia but not frankly malignant disease or metastases. The disease was produced at a high frequency when 10-day-old embryos were infected. Breast muscle (pectoralis major and minor) was affected with variable severity in all birds, thigh muscles were affected occasionally, while other thoracic, external abdominal, and wing muscles were affected very rarely. Progression of proliferative muscle lesions was demonstrated by sequential necropsies and microscopic examination of muscle samples. The disease appeared before 2 weeks of age, and thin white streaks with minimal cellular proliferation were the earliest lesions observed. Elongated, spindle-shaped tumors were the most advanced form of proliferation observed. The advanced lesions showed cellular anaplasia and signs of rapid growth, and appeared most commonly at 6 weeks of age or later. Histologically, mild proliferation correlated with normal appearing fibroblasts producing collagen. Severe proliferation correlated with anaplastic, rapidly dividing cells producing little collagen and a high mitotic index. A decrease in the virus dose resulted in less severe fibromatosis, but at least one chicken infected with a low virus dose showed severe disease. When 10- or 35-day-old hatched chicks were injected in the breast muscle with EU-8 or K-3, a low incidence of lymphoid leukosis was observed, but no fibromatosis resulted during an observation period of 27 weeks. Infectious virus was extracted from the fibromatosis in the breast muscle, and extracted virus caused the same disease when injected in 10-day-old embryos. No transforming activity was observed on normal chick embryo fibroblast monolayers. Cell cultures were established from areas in the breast muscle affected by fibromatosis, and from discrete tumors. These cells did not show any signs of transformation in culture. Supernatants obtained from these cell cultures caused fibromatosis when injected into 10-day old embryos, but did not transform normal chick embryo fibroblasts. PMID- 3030544 TI - Small cell lung cancer cell line derived from a primary tumor with a characteristic deletion of 3p. AB - Chemotherapy plus surgery is feasible and potentially effective in selected patients with small cell lung cancer (SCLC) and provides a unique opportunity to study SCLC early in its biological history. The in vitro characteristics of a SCLC cell line derived from a resected lung primary tumor after treatment with 3 courses of chemotherapy is described. The original SCLC cell line UMC-SCLC-1 exhibited features of classic SCLC with typical morphology and growth characteristics, high levels of dopa decarboxylase, bombesin-like peptides, neuron-specific enolase and calcitonin, and the presence of neurosecretory granules and demonstrated the deletion of the short arm of chromosome 3. After multiple passages, UMC-SCLC-1 gradually changed its culture characteristics to a cell line, UMC-SCLC-1A, with morphological features of large cell anaplastic carcinoma, an altered growth pattern, decrease in calcitonin, and increase in radioresistance but retained the other biochemical markers of classic SCLC (bombesin and dopa decarboxylase production). Serial DNA content analyses showed that increased aneuploidy during continuous culture in vitro was associated with the morphological changes. Both UMC-SCLC-1 and UMC-SCLC-1A demonstrated the deletion of chromosome 3p, amplification and abundant expression of N-myc, and increased expression of c-raf. Chemotherapy sensitivities were stable throughout multiple passages and correlated with in vivo response. UMC-SCLC-1A represents a unique SCLC cell line with heterogeneous properties of both classic and morphological variant SCLC cell lines. In addition, the characteristic deletion of 3p, previously described in cultures derived from metastatic lesions and heavily pretreated patients, is seen in a primary lesion early in the natural history of SCLC. PMID- 3030545 TI - Macrophage potentiation of invasive capacity of rat ascites hepatoma cells. AB - In vitro and in vivo invasive capacities of rat ascites hepatoma cells (AH 130) that had been cultured on the feeder layers of rat macrophages were examined. The in vitro invasive capacity of the tumor cells was measured by their ability to form tumor cell colonies underneath cultured mesothelial cell monolayers; in vivo invasive capacity was examined by the implantation of the tumor cells into the rat peritoneal cavity. When the tumor cells were precultured on a macrophage feeder layer, the in vitro invasive capacity of the tumor cells increased almost 10 times as much as that of uncocultred control cells. The cocultured tumor cells, when implanted in rat peritoneal cavity, infiltrated extensively in the peritoneum and formed many tumor nodules and enlarged metastatic lymph nodes. Implantation of the uncocultured tumor cells did not develop any macroscopically detectable nodules. The effect of macrophages was reversed by subculturing the cocultured tumor cells without macrophages. Treatment of the tumor cells with the medium conditioned by macrophage culture did not result in the increase in invasive capacity. Almost 50% of the macrophage-mediated enhancement of the in vitro invasive capacity was inhibited by the simultaneous addition of superoxide dismutase and catalase at the time of tumor cell-macrophage coculture. PMID- 3030546 TI - In vivo administration of purified human interleukin-2 to patients with cancer: development of interleukin-2 receptor positive cells and circulating soluble interleukin-2 receptors following interleukin-2 administration. AB - Recent studies have demonstrated efficacy of immunotherapies including interleukin-2 (IL-2) in the treatment of malignancies in rodents and humans. High levels of IL-2 receptor-positive cells were found in the peripheral blood of patients receiving recombinant IL-2 in these Phase I clinical trials. This was demonstrated both in patients receiving i.v. IL-2 who had detectable circulating levels of IL-2 as well as in patients receiving i.p. IL-2 who did not. Up to 100% of the anti-Tac binding could be inhibited by preincubation with IL-2 indicating that this was indeed an IL-2 receptor that was identified. Two-color experiments demonstrated that few Leu 2-positive cells (less than 5-10%) but over 30% of the Leu 3-positive cells bore Tac antigen. Most of the M3-positive monocytes were Tac positive (83.7%) and negative for other T-cell (Leu-4) and nonspecific murine markers (Lyt-2 and Thy 1.2). Although normal individuals had a mean of only 186 units/ml (range, 83-335 units/ml) of soluble IL-2 receptor, patients receiving IL 2 had as much as 20,000 units/ml of soluble IL-2 receptor line in their serum. The physiological role of the IL-2 receptor identified on the cell surface of Leu 3 and M3-positive cells as well as in the serum is unclear. Soluble IL-2 receptors appeared in the circulation early following IL-2 administration, approximately 1 week prior to the detection of circulating IL-2 receptor-bearing cells. Further studies will be needed to assess the role of IL-2 in monocyte function, the precise function of IL-2 receptor-bearing Leu 3-positive cells, and the relationship of these findings to the toxicity and success of this immunotherapy in humans. PMID- 3030547 TI - High-dose cytarabine in small cell lung cancer. PMID- 3030548 TI - "Ifosfamide plus N-acetylcysteine in the treatment of small cell and non-small cell carcinoma of the lung: a Southeastern Cancer Study Group Trial". PMID- 3030549 TI - [Left ventricular hypertrophy: structural, electrophysiologic and biochemical changes at the cellular level]. PMID- 3030550 TI - Studies on the interrelationship between hormonal imprinting and membrane potential in model experiments on Tetrahymena. PMID- 3030551 TI - Effect of progesterone on the sodium transporting mechanism of isolated toad skin. PMID- 3030552 TI - Oxidoreductase distribution in the fat body and symbionts of the German cockroach Blattella germanica: a histochemical approach. PMID- 3030553 TI - [Morphophysiologic disorders after adrenalectomy in the rabbit]. PMID- 3030554 TI - Enkephalin in the goldfish retina. AB - Enkephalin-like immunoreactive amacrine cells were visualized using the highly sensitive avidin-biotin method. The somas of these cells were situated in the inner nuclear and ganglion cell layers. Enkephalin-stained processes were observed in layers 1, 3, and 5 of the inner plexiform layer. The biosynthesis of sulfur-containing compounds in the goldfish retina was studied by means of a pulse-chase incubation with 35S-methionine. A 35S-labeled compound, which comigrated with authentic Met5-enkephalin on high-performance liquid chromatography (HPLC), was synthesized and was bound competitively by antibodies to enkephalin and by opiate receptors. This compound was tentatively identified as "Met5-enkephalin." The newly synthesized 35S-Met5-enkephalin was released upon depolarization of the retina with a high K+ concentration. This K+-stimulated release was greatly suppressed by 5 mM Co2+, suggesting that the release was Ca2+ dependent. Using a double-label technique, enkephalin immunoreactivity and gamma aminobutyric acid (GABA) uptake were colocalized to some amacrine cells, whereas others labeled only for enkephalin or GABA. The possible significance of enkephalin-GABA interactions is also discussed. PMID- 3030555 TI - Effect of retinoic acid on nerve growth factor receptors. AB - Retinoic acid (RA), a naturally occurring metabolite of vitamin A, increased the number of receptors for nerve growth factor (NGF) in cultured human neuroblastoma cells (LA-N-1), as indicated by an immunofluorescence assay of cell surface receptors and by specific binding of 125I-NGF to solubilized receptors. Analysis of 125I-NGF binding showed that RA increased the number of both high affinity and low affinity receptors for NGF without affecting the equilibrium dissociation constants. Neurite outgrowth similar to that produced by NGF occurred following RA-treatment in LA-N-1 cells, in the SY5Y subclone of SK-N-SH human neuroblastoma cells and in explanted chick dorsal root ganglia (DRG). Whether morphological changes following RA treatment are directly related to the increase in NGF receptors is unknown. Data presented here are consistent with literature reports that RA modifies cell surface glycoproteins, including those that act as cell surface receptors for epidermal growth factor and insulin. PMID- 3030556 TI - The yeast polyubiquitin gene is essential for resistance to high temperatures, starvation, and other stresses. AB - Conjugation of ubiquitin to intracellular proteins mediates their selective degradation in eukaryotes. In the yeast Saccharomyces cerevisiae, four distinct ubiquitin-coding loci have been described. UBI1, UBI2, and UBI3 each encode hybrid proteins in which ubiquitin is fused to unrelated sequences. The fourth gene, UBI4, contains five ubiquitin-coding elements in a head-to-tail arrangement, and thus encodes a polyubiquitin precursor protein. A precise, oligonucleotide-directed deletion of UBI4 was constructed in vitro and substituted in the yeast genome in place of the wild-type allele. ubi4 deletion mutants are viable as vegetative cells, grow at wild-type rates, and contain wild type levels of free ubiquitin under exponential growth conditions. However, although ubi4/UBI4 diploids can form four initially viable spores, the two ubi4 spores within the ascus lose viability extremely rapidly, apparently a novel phenotype in yeast. Furthermore, ubi4/ubi4 diploids are sporulation-defective. ubi4 mutants are also hypersensitive to high temperatures, starvation, and amino acid analogs. These three conditions, while diverse in nature, are all known to induce stress proteins. Expression of the UBI4 gene is similarly induced by either heat stress or starvation. These results indicate that UBI4 is specifically required for the resistance of cells to stress, and that ubiquitin is an essential component of the stress response system. PMID- 3030557 TI - KAR1, a gene required for function of both intranuclear and extranuclear microtubules in yeast. AB - Molecular analysis of the KAR1 gene of yeast has shown that it is required for both mitosis and conjugation. The gene was originally identified by mutations that prevent nuclear fusion. By in vitro mutagenesis and gene replacement we have demonstrated that the gene is an essential cell division cycle gene. Temperature sensitive mutant strains show defects in spindle pole body duplication and chromosome disjunction. Overproduction of the gene product blocks spindle pole body duplication, producing a cell cycle arrest phenotype similar to that of the Kar- temperature-sensitive mutations. Long, aberrant extranuclear microtubules are formed in the temperature-sensitive mutants arrested at the nonpermissive temperature as well as in kar1-1 during conjugation. These observations suggest that the KAR1 gene is required for the normal function of both the intranuclear and extranuclear microtubules. PMID- 3030558 TI - 42 bp element from LDL receptor gene confers end-product repression by sterols when inserted into viral TK promoter. AB - The LDL receptor, which mediates the cellular uptake of cholesterol, is subject to classic end-product repression when cholesterol accumulates in the cell. We here show that the sensitivity to end-product repression depends upon a 42 bp element in the 5'-flanking region of the human LDL receptor gene. This sequence, designated sterol regulatory element 42 (SRE 42), contains two 16 bp direct repeats that exhibit positive and negative transcriptional activities. Cells transfected with a fusion gene containing SRE 42 inserted into the promoter of the herpes simplex viral TK gene produced abundant mRNA when grown without sterols. When sterols were present, the mRNA was reduced by 57%-95%, depending on the number of copies of SRE in the fusion gene. These transfection data plus DNAase I footprinting experiments suggest a model of end-product repression in which the end product (sterols) opposes the action of a positive transcription factor that binds to a discrete promoter element. PMID- 3030559 TI - Regulation of transmembrane signaling by receptor phosphorylation. AB - At least two major effects of receptor phosphorylation have been identified- regulation of receptor function, and regulation of receptor distribution. In many cases where phosphorylation directly alters the functions of receptors, this appears to be in a negative direction. Such decreases in receptor activity may reflect reduced ability to interact with biochemical effectors (e.g., the beta adrenergic receptor, rhodopsin), reduced affinity for binding agonist ligands (EGF,IGF-I, insulin receptors) or reduced enzymatic activity (e.g., tyrosine kinase activity of the insulin or EGF receptor). In all instances, these negative modulations are associated with phosphorylation of serine and/or threonine residues of the receptor proteins. In contrast, the tyrosine kinase receptors also appear to be susceptible to positive modulation by phosphorylation. With these receptors, autophosphorylation of tyrosine residues may lead to enhanced protein-tyrosine kinase activity of the receptors and increased receptor function. In addition, the subcellular distribution of a receptor may be regulated by its phosphorylation status (e.g., the beta-adrenergic receptor, receptors for insulin, EGF, IGF-II, and transferrin). The emerging paradigm is that receptor phosphorylation may in some way promote receptor internalization into sequestered compartments where dephosphorylation occurs. The molecular and cellular mechanisms involved in translating changes in receptor phosphorylation into changes in receptor distribution remain to be elucidated. Moreover, the biological role of receptor internalization may be quite varied. Thus, in the case of the beta-adrenergic receptor, it may serve primarily as a mechanism for bringing the phosphorylated receptors into contact with intracellular phosphatases that dephosphorylate and resensitize it. By contrast, for the transferrin receptor and other receptors involved in receptor-mediated endocytosis, the internalization presumably functions to carry some specific ligand or metabolite into the cell. The role of phosphorylation in regulating receptor function dramatically extends the range of regulatory control of this important covalent modification. PMID- 3030560 TI - The unusual conformation adopted by the adenine tracts in kinetoplast DNA. AB - To determine the structural features responsible for the curvature of kinetoplast DNA, we studied 13 adenine tracts in Crithidia fasciculata kinetoplast DNA. The structures of the A tracts were analyzed by cutting the DNA with hydroxyl radical. Reactivity of hydroxyl radical toward the DNA backbone progressively decreased in the 5'----3' direction of each A tract. The cutting pattern of the T rich strand was offset by 1 or 2 bp from the pattern on the A-rich strand. An A tract in a restriction fragment from plasmid pBR322 had the same cutting pattern as the kinetoplast A tracts. We interpret these experiments to show that in A tracts the width of the minor groove decreases smoothly from the 5'----3' end of the A tract. PMID- 3030561 TI - Transcription termination factor rho is an RNA-DNA helicase. AB - E. coli rho factor can unwind a short RNA-DNA duplex in vitro. The duplex is formed between a polylinker sequence at the 3' end of RNA derived from the rho dependent terminator trp t' and the complementary sequence in a single-strand DNA molecule. Release of trp t' RNA from the duplex requires nucleoside triphosphate hydrolysis by rho's NTPase activity and is dependent on rho recognition of the RNA that is 5' to the RNA-DNA duplex region. The direction of helix unwinding appears to be 5' to 3' along the RNA molecule. These characteristics now account for how the RNA-binding and RNA-dependent NTP hydrolysis activities of rho may participate directly in transcription termination. Our results suggest that NTP hydrolysis is utilized to help unwind the RNA-DNA duplex at the 3' end of a nascent transcript, facilitating RNA release from the DNA template. PMID- 3030562 TI - A tail of two src's: mutatis mutandis. PMID- 3030563 TI - The gamma delta resolvase induces an unusual DNA structure at the recombinational crossover point. AB - gamma delta resolvase, a transposon-encoded protein active in site-specific recombination, induces a structural change in the DNA at the recombinational crossover point that results in enhanced intercalation of the foot-printing reagent MPE X Fe(II). The structural change correlates with the formation of a bend in the DNA: a mutant resolvase that binds to the crossover site but induces little or no bend does not promote intercalation. The properties of the mutant protein suggest that the induced structural change, which we propose is a localized kink, is required for recombination. PMID- 3030564 TI - The functions and relationships of Ty-VLP proteins in yeast reflect those of mammalian retroviral proteins. AB - We have identified the major structural core proteins of Ty virus-like particles (Ty-VLPs) and shown that they are generated by proteolytic cleavage of the primary translation product of TYA, p1. This precursor protein is therefore functionally similar to the gag precursor of retroviruses. Cleavage is mediated by a Ty-encoded protease located at the 5' region of TYB and is accompanied by a change in particle morphology. p1 contains sufficient information for the assembly of a pre-Ty-VLP complex, which does not require the presence of either Ty protease or reverse transcriptase. The results indicate that the requirements and pathway of Ty-VLP formation reflect the initial stages of mammalian retroviral assembly and further support the idea of a common origin for Ty elements and retroviruses. PMID- 3030565 TI - Yeast HAP1 activator binds to two upstream activation sites of different sequence. AB - We show that the HAP1 protein binds in vitro to the upstream activation site (UAS) of the yeast CYC7 gene. Strikingly, this sequence bears no obvious similarity to the sequence bound by HAP1 at UAS1 of the CYC1 gene. The CYC1 and CYC7 sites compete for binding to HAP1 and have comparable affinities for the protein. The gross features of the interaction of HAP1 with the two sites are similar: multiple major and minor groove contacts, spanning 23 bp, on one helical face, with a back-side major groove contact toward one end. The precise positions of the contacts differ, however. A mutant form of HAP1, HAP1-18, abolishes the ability of the protein to bind to UAS1 but not CYC7 DNA. Possible mechanisms for how a single protein recognizes two sequences are discussed. PMID- 3030566 TI - Regulation of the human interleukin-2 receptor alpha chain promoter: activation of a nonfunctional promoter by the transactivator gene of HTLV-I. AB - We have characterized regulatory regions of the human IL-2 receptor alpha chain (IL2R alpha) promoter. 5' deletion constructs extending to -327 directed CAT expression in HTLV-I-infected T cells, which express IL2R alpha constitutively, and in Jurkat cells, which express IL2R alpha only after induction. Deletions to 267 and -265 were active only in HTLV-I-transformed T cells, but their activity in Jurkat cells was restored by cotransfection of a construct expressing the HTLV I transactivator protein (tat-I). However, HTLV-I-infected human osteosarcoma cells do not express IL2R alpha-CAT constructs. Thus cell-type-specific factors are required for IL2R alpha expression, and direct or indirect interaction(s) between tat-I and a specific region of the IL2R alpha promoter may cause altered regulation. Tat-I also augments IL2-CAT expression under some conditions, suggesting possible autocrine or paracrine mechanisms for HTLV-I-induced leukemogenesis. PMID- 3030567 TI - Yeast HAP1 activator competes with the factor RC2 for binding to the upstream activation site UAS1 of the CYC1 gene. AB - We show that the yeast HAP1 activator locus encodes a protein that binds in vitro to the upstream activation site, UAS1, of the CYC1 gene (iso-1-cytochrome c). Binding of wild-type HAP1 and truncated HAP1 derivatives to UAS1 is evident in crudely fractionated yeast extracts using the gel electrophoresis DNA binding assay. The binding of HAP1 in vitro, like the activity of UAS1 in vivo, is stimulated by heme. HAP1 binds to region B, one of two portions of UAS1 shown to be important by genetic analysis of the site. Surprisingly, HAP1 binds to the same sequence as a second factor, RC2. Both HAP1 and RC2 bind to the same side of the helix, and make similar but not identical major and minor groove contacts that span two full turns. An additional factor that binds to the second important part of UAS1, the region A factor (RAF), is also identified. A model depicting the interplay of HAP1, RC2, and RAF in the control of UAS1 is presented. PMID- 3030569 TI - DNA breakdown of 3T3 and SV 40-transformed 3T3 cells cultured in suspension. AB - It was examined what effect of suspension culture exerted on prelabeled DNA of 3T3 and SV 40 transformed cells (SV3T3). On an alkaline sucrose density gradient the small size DNA of 3T3 cells increased with time of suspension, while that of SV3T3 did not. Furthermore, it was demonstrated that prelabeled DNA of suspended 3T3 cells became small on a neutral sucrose density gradient, in an alkaline and a neutral elution. When SV3T3 cells were treated with dimethylsulfoxide, the smaller DNA appeared on an alkaline sucrose density gradient. PMID- 3030568 TI - Creation of a processed pseudogene by retroviral infection. AB - We have previously characterized a cell line transformed by a Rous sarcoma virus mutant, SE21Q1b, which contains a mutation preventing encapsidation of genomic RNA. A unique property of this mutant is that cellular RNAs are packaged into virions, even in the presence of replication-competent virus. In the current study, SE21Q1b quail cells were transfected with the plasmids pRSVneo or pCMVneo. Virions produced by SE21Q1b neoR clones contained neo RNA, and when virus from some SE21Q1b neoR clones was used to infect a chemically transformed quail cell line, QT35, neoR QT35 clones were obtained that contained single integrated copies of the neo gene. An intron inserted into pRSVneo was removed during gene transfer. These data are consistent with transfer of neo mRNAs by a pathway involving reverse transcription of mRNA encapsidated within SE21Q1b virions, and integration of resultant cDNAs into the genome of infected QT35 cells. PMID- 3030570 TI - Cardiac effects of menadione and its semiquinone. PMID- 3030571 TI - Positioning of nucleosomes reconstituted with xeroderma pigmentosum and normal histones. AB - Positioning of nucleosomes was examined in a reconstituted system using a plasmid DNA and histones from normal human and xeroderma pigmentosum, complementation group A (XPA), lymphoblastoid cells. The present studies indicate that the arrangement of nucleosomes, composed of normal human histones, in a region near the SV40 origin of replication on the plasmid DNA, is nonrandom. The alignment of nucleosomes in this region was not affected by the presence of histone H1. No difference in nucleosome positioning was observed when the nucleosomes were composed of histones from XPA cells. PMID- 3030572 TI - Electron spin resonance and pulse radiolysis studies on the spin trapping of sulphur-centered radicals. AB - Thiyl radicals are shown to be readily trapped with the spin traps 5,5-dimethyl-1 pyrroline-N-oxide (DMPO) and 3,3,5,5-tetramethyl-1-pyrroline-N-oxide (TMPO) giving characteristic spin adducts with hyperfine coupling constants aN 1.52 1.58, aH 1.52-1.80 mT, and g values in the range 2.0065-2.0067 for the DMPO adducts and aN 1.50-1.56, aH 1.70-1.92 mT, g 20049-2.0051 for the TMPO adducts. Kinetic data obtained from pulse radiolysis studies show that, in general, thiyl radicals react rapidly with these spin traps with rate constants of the order of 10(7)-10(8) dm3 mol-1 s-1. The tetramethylated spin trap TMPO though giving slightly less intense electron spin resonance (ESR) spectra, produces longer lived adducts, and is suggested to be of greater utility due to the more characteristic nature of the coupling constants of the observed adducts; reaction of certain thiyl radicals with DMPO produces adducts which are superficially similar to the hydroxyl radical adduct to the same trap. PMID- 3030574 TI - Studies on peptides. CXLIV. Synthesis and immunological properties of formylmethionyl human adrenocorticotropin. PMID- 3030573 TI - Allylic compounds bind directly to DNA: investigation of the binding mechanisms in vitro. AB - The in vitro binding of three allyl compounds, allyl methanesulphonate, allyl bromide and allyl chloride to DNA from salmon sperm was investigated. Kinetic DNA alkylation measurements revealed significant differences between the strongly alkylating allyl methanesulphonate and allyl bromide with half-life values of 1.5 h and 8.1 h, respectively and the weakly alkylating allyl chloride with a half life value of 360 h. Five alkylated nucleic bases, 3-allyladenine, N6 allyladenine, N2-allylguanine, 7-allylguanine and O6-allylguanine, could be identified after DNA-alkylation with all three allyl compounds. PMID- 3030575 TI - Synthesis and pharmacological properties of morphine congeners having pendant crown ether as an opioid receptor probe. PMID- 3030576 TI - Recent advances in identifying the functions of gangliosides. AB - The recent development of several new approaches has proven extremely useful in identifying functions for gangliosides, the sialic-acid containing glycosphingolipids. The first is the incorporation of exogenous gangliosides into the plasma membrane of ganglioside-deficient cells. Using this approach, specific gangliosides have been identified as the receptors for certain bacterial toxins and viruses and as important factors in the organization of fibronectin into an extracellular matrix. The second approach has been a ligand blotting technique which allows detection of ganglioside-binding proteins such as toxins and antibodies. Gangliosides are separated by thin-layer chromatography and overlain with the protein of interest. Specific binding of the ligand to gangliosides can then be detected by either direct or indirect methods. The third approach is the use of the B or binding subunit of cholera toxin as a specific probe for endogenous plasma membrane ganglioside function. The ability of the B subunit to alter the growth of cells directly demonstrates a role for gangliosides as biotransducers of signals for the regulation of cell growth. PMID- 3030577 TI - Immunobiology of complete molar pregnancy and gestational trophoblastic tumor. AB - The unique curability of gestational trophoblastic tumors may in part be attributable to a host immunologic response. The occurrence of rapidly progressive and fatal choriocarcinoma may be favored by histocompatibility between patients and their partners. However, histocompatibility is not a prerequisite for the development and persistence of gestational choriocarcinoma. The expression of HLA by choriocarcinoma cells in culture is enhanced following incubation with gamma-interferon and this may be of both biologic and clinical significance. Complete molar pregnancy is a complete allograft because all molar chromosomes are of paternal origin. Patients with complete mole are sensitized to paternal HLA antigen which is expressed in molar tissue. Other polymorphic antigen systems including trophoblast-leukocyte common antigens and placental type alkaline phosphatase are also expressed in molar tissue. We have studied the immunopathology of the molar implantation site to investigate possible humoral and cellular immune responses. The relationships among normal placenta, complete mole and choriocarcinoma are not clearly understood. The pattern of expression of oncofetal antigens in these three gestational tissues may be used to assess trophoblastic differentiation. In studies to date, molar trophoblast has the same pattern of expression of oncofetal antigens as normal placental trophoblast. We will review recent advances in our understanding of the immunobiology of gestational trophoblastic disease and suggest new directions for further research. PMID- 3030579 TI - Metaphase and anaphase analysis of V79 cells exposed to erionite, UICC chrysotile and UICC crocidolite. AB - The cytogenetic effects of erionite treatment of V79 cells were compared with those of UICC crocidolite and UICC chrysotile treatment. A significant reduction in diploid cells with an accompanying increase in aneuploid and polyploid cells was observed with all three treatments. In the erionite-treated cultures, an increase in aneuploidy was observed at all dose levels ranging from 10 to 100 micrograms/ml, whereas in the crocidolite- and chrysotile-treated cultures, significant increases in aneuploidy were observed at all dose levels except the low dose, 10 micrograms/ml. Chromatid aberrations were observed in cultures treated with crocidolite and chrysotile and were especially pronounced at dose 100 micrograms/ml of chrysotile. The clastogenic effect of erionite was weaker but statistically significant at dose 100 micrograms/ml. An extrapolation of these cytogenetic changes over dose in number of fibers suggests that erionite was more reactive than the other two minerals in producing aneuploidy. The number of fibers required to produce a similar degree of cytogenetic effects was several orders of magnitude higher for chrysotile and crocidolite than erionite. These results correlate with the higher tumorigenic potency of erionite. In general, fewer cells treated with erionite entered anaphase than those treated with the other two minerals. As a result, abnormal anaphases representing chromosomal mis segregation were observed only in the chrysotile- and crocidolite-treated cultures. To our knowledge, this is the first report on cytogenetic effects of erionite. PMID- 3030580 TI - Noninvasive detection of rejection of transplanted hearts with indium-111-labeled lymphocytes. AB - To determine whether cardiac transplant rejection can be detected noninvasively with indium-111 (111In)-labeled lymphocytes, we studied 11 dogs with thoracic heterotopic cardiac transplants without immunosuppression and five dogs with transplants treated with cyclosporine (10 mg/kg/day) and prednisone (1 mg/kg/day). All were evaluated sequentially with gamma scintigraphy after administration of 150 to 350 muCi of autologous 111In-lymphocytes. Technetium-99m labeled red blood cells (1 to 3 mCi) were used for correction of radioactivity in the blood pool attributable to circulating labeled lymphocytes. Lymphocyte infiltration was quantified as the ratio of indium in the myocardium of the transplant or native heart compared with that in blood (indium excess, IE). Results were correlated with mechanical and electrical activity of allografts and with histologic findings in sequential biopsy specimens. In untreated dogs (n = 11), IE was 15.5 +/- 7.0 (SD) in transplanted hearts undergoing rejection and 0.4 +/- 1.1 in native hearts on the day before animals were killed (p less than .01). In dogs treated with cyclosporine and prednisone (n = 5), IE was minimal in allografts during the course of immunosuppression (0.8 +/- 0.4) and increased to 22.9 +/- 11.1 after immunosuppression was stopped. Scintigraphic criteria of rejection (IE greater than 2 SD above that in native hearts) correlated with results of biopsies indicative of rejection and appeared before electrophysiologic or mechanical manifestations of dysfunction. Thus infiltration of labeled lymphocytes in allografts, indicative of rejection, is detectable noninvasively by gamma scintigraphy and provides a sensitive approach potentially applicable to clinical monitoring for early detection of rejection and guidance for titration of immunosuppressive measures. PMID- 3030581 TI - Prevention of fog-induced bronchospasm by nedocromil sodium. AB - A single dose double-blind crossover study was performed to compare the efficacy of nedocromil sodium (4 mg) and placebo administered from pressurized aerosols against bronchoconstriction induced by the inhalation of ultrasonically nebulized distilled water (fog) in twelve asthmatic subjects. Neither active nor placebo pre-treatment produced any significant change in baseline FEV1 and SRaw. Nedocromil sodium significantly attenuated fog-induced falls in FEV1 (P less than 0.001) and increased specific airways resistance (SRaw, P less than 0.01). The results provide further evidence of the potential therapeutic usefulness of nedocromil sodium in the management of chronic obstructive airways disease. PMID- 3030582 TI - Direct determination of angiotensin-converting enzyme inhibitors in plasma by radioenzymatic assay. AB - We describe a simple, rapid, specific radioenzymatic assay for "CGS 16617," a new, potent inhibitor of angiotensin-converting enzyme (ACE; EC 3.4.15.1) in human plasma. This assay is based on the principle that the inhibitor (i.e., the drug) binds to the ACE in plasma and hence the amount of free ACE in plasma is inversely related to the amount of active inhibitor present. Free enzyme is reacted with a radiolabeled substrate, and the radioactive product is selectively extracted into the scintillation cocktail for quantification. Fivefold-diluted plasma samples are incubated with [3H]hippuryl-glycyl-glycine enzyme substrate at 37 degrees C for 30 min and the liberated [3H]hippuric acid is selectively extracted into scintillation cocktail. The radioactivity is counted in a liquid scintillation counter. Both within-run and between-run, the variability (CV) of the assay is less than 10%. As little as 200 ng of the drug per liter can be quantified in 50-microL plasma samples. The method can also be used to assay two other ACE inhibitors, pentopril and CGS 14831, demonstrating that the method can be readily adapted to any ACE inhibitor having a single active component in plasma. The ester prodrug pentopril can also be assayed after ester hydrolysis. This method is suitable for analysis of large numbers of samples in clinical laboratories for routine monitoring of the concentrations of active ACE inhibitors in blood. PMID- 3030583 TI - Low A-esterase activity in serum of patients with fish-eye disease. AB - The activity of HDL-associated paraoxonase in the lipoprotein fraction of serum from two patients with fish-eye disease (FED) was only 11% of the mean value for control subjects. Similarly, concentrations of apolipoproteins A-I and A-II in the serum of FED subjects were only 10% of those in normal subjects. Thus the ratio of enzyme activity to A-I and A-II is virtually the same in these two groups of subjects. This suggests that paraoxonase activity of the lipoprotein fraction is associated with one or both of these apolipoproteins. PMID- 3030584 TI - Comparative measurements for determination of alkali ions in human blood serum by ion-chromatography. PMID- 3030585 TI - The management of glomus jugulare tumours. AB - The treatment details of 58 patients treated for glomus jugulare tumours in Newcastle upon Tyne are examined in the light of other studies reported in the literature. For the group of 55 patients treated by radiotherapy, the 20 year survival is 94% (determined actuarially). The 20 year disease-free survival (determined actuarially) is 77%. This is comparable with other series reported. As no glomus tympanicum tumour has recurred following surgery and there has been no morbidity due to these tumours they have not been included in the series. It is recommended that patients who are fit and have tumours confined to the tympanum should have primary surgical treatment. All other patients should be treated by accurately planned radiotherapy, using a dose of 50Gy in 5 weeks to the tumour volume. The morbidity of this treatment policy will be low. PMID- 3030586 TI - Persistent effects on blood pressure and renal haemodynamics following chronic angiotensin converting enzyme inhibition with perindopril. AB - Spontaneously hypertensive rats (SHR) and Wistar-Kyoto (WKY) rats aged 4 and 16 weeks were given an acute oral dose of either Perindopril (3 mg/kg) or vehicle. Direct blood pressure (BP), glomerular filtration rate (GFR) and renal blood flow (RBF) were measured, and renal vascular resistance (RVR) calculated. GFR and RBF were lower in vehicle-treated SHR than WKY at 4 weeks of age, but were not different at 16 weeks. Acute Perindopril increased GFR and RBF and reduced RVR in both strains at both 4 and 16 weeks. Total body sodium, sodium intake and blood pressure were measured in SHR and WKY from 1 to 28 weeks of age. Rats of both strains were treated daily between 4 and 16 weeks of age with either Perindopril (3 mg/kg per day) or vehicle. Chronic Perindopril treatment prevented the development of hypertension in the SHR. From 16 to 28 weeks of age, after stopping Perindopril, BP rose slowly in SHR, but remained lower than vehicle treated SHR. No changes in total body sodium occurred during Perindopril treatment. GFR and RBF were measured in SHR and WKY chronically treated with either Perindopril or vehicle, 3 days or 12 weeks after stopping treatment. In WKY, GFR and RBF were not different between Perindopril-treated and untreated rats at either measurement. In SHR, GFR and RBF remained significantly higher in rats previously treated with Perindopril at both ages. These findings suggest that renal haemodynamic abnormalities may be important in the initiation of hypertension in the SHR. These renal circulatory abnormalities and the hypertension of the SHR depend, at least in part, on intact converting enzyme activity, yet appear to be independent of abnormalities of total body sodium. At a later age, hypertension seems to develop independently of renal vascular abnormalities. PMID- 3030587 TI - Aldosterone responses to ACTH during hypoxia in conscious rats. AB - This study examined the aldosterone response to physiological ACTH infusion (4 ng/kg per min) and pharmacological ACTH injection (1.8 microgram) in conscious Long-Evans rats at 42 h of normoxia (21% O2) or hypoxia (10% O2). Hypoxia per se (no exogenous ACTH) significantly decreased control aldosterone levels despite elevated endogenous plasma ACTH). Hypoxia attenuated the aldosterone responses to ACTH infusion but not to ACTH injection. It was concluded that hypoxia attenuates aldosterone responses to small increases in ACTH, and large increases in ACTH can override this apparent decrease in adrenocortical sensitivity. PMID- 3030588 TI - O-phenylenediamine oxidation by phorbol myristate acetate-stimulated human polymorphonuclear leukocytes: characterization of two distinct oxidative mechanisms. AB - O-Phenylenediamine (OPD) oxidation has been extensively utilized for the measurement of peroxidase-mediated catabolism of hydrogen peroxide. However, until now this system has not been evaluated for the measurement of hydrogen peroxide produced upon activation of the hexose monophosphate shunt (HMPS) in polymorphonuclear leukocytes (PMNs). OPD oxidation by phorbol myristate acetate (PMA)-stimulated PMNs was mediated by both hydrogen peroxide and superoxide produced by the activation of the HMPS. Furthermore, OPD oxidation by an oxidative mechanism independent of the HMPS was observed by the PMA stimulation of PMNs obtained from patients with chronic granulomatous disease (CGD). This HMPS-independent OPD oxidation was inhibited by superoxide dismutase or 1 mM potassium cyanide (KCN). Superoxide dismutase, catalase, or 1 mM potassium cyanide inhibited 50% OPD oxidation obtained with PMA-stimulated normal PMNs. PMA treatment of purified human myeloperoxidase (MPO) produced OPD oxidation which is inhibited by superoxide dismutase or 1 mM KCN. These data indicate that OPD oxidation observed with CGD PMNs is mediated by a PMA-induced oxidase activity of myeloperoxidase. OPD oxidation in the presence of 1 mM KCN is a method comparable in sensitivity with ferricytochrome c reduction for the evaluation of HMPS activity. Furthermore, the OPD assay can measure myeloperoxidase oxidase activity in PMA-stimulated PMNs. PMID- 3030589 TI - Hepatic reticuloendothelial system activation in autoimmune mice: differences between (NZB X NZW)F1 and MRL-lpr/lpr strains. AB - 5'-Nucleotidase (5'N) activity and Ia expression of hepatic nonparenchymal cells (NPCs) of the autoimmune (NZB X NZW)F1 and MRL-lpr/lpr and nonautoimmune C3H/FeJ, DBA/2J, and A/J strains were assayed to determine endogenous activation of the hepatic reticuloendothelial system (RES). Pretreatment of the nonautoimmune strains with Corynebacterium parvum resulted in decreases in Fc receptor expressor and 5'N and an increase in Ia expression of NPCs. Endogenous hepatic RES activation, as measured by low 5'N and high Ia expression, was observed in both the autoimmune MRL-lpr/lpr and (NZB X NZW)F1 strains even without C. parvum pretreatment. However, in the MRL-lpr/lpr strain, age is a dependent variable in activation and correlates with delayed clearance from the circulation of soluble IgG immune complexes. In the (NZB X NZW)F1 strain, the observation of decreased 5'-nucleotidase activity and increased percentage of Ia-positive cells at all ages suggests a primary abnormality. Therefore, different genetic or exogenous factors may be responsible for the activation of the hepatic RES in these autoimmune strains. PMID- 3030590 TI - Cellular antigens in nephroblastoma: identification with monoclonal antibodies which recognize hemopoietic cells. AB - The correlation between expression of differentiation antigens and morphogenesis was examined in 11 human nephroblastomas using monoclonal antibodies which recognize both hemopoietic and renal cells. Analysis of frozen tissue sections by indirect immunofluorescence revealed that blastema cells and some tubules were recognized by BA-1 and OKB2, which identify unstimulated B cells and granulocytes. Stroma, some tubules, and focal blastema were recognized by BA-2, which identifies a 24-kDa antigen on leukemic cells and platelets. Mature epithelium in glomerular bodies was identified by C3bR and J5, which recognize CR1 and the common acute lymphoblastic leukemia antigen, respectively. Tissue sections from a clear cell sarcoma and a mesoblastic nephroma were notably reactive only with BA-2. These observations demonstrate that the relationship between antigen expression and morphology in nephroblastomas is similar to that observed in fetal kidney and suggest that expression of these cell surface antigens may have a role in morphogenesis. PMID- 3030591 TI - Heart-specific autoantibodies induced by Coxsackievirus B3: identification of heart autoantigens. AB - Postinfection sera from A.CA/SnJ A.SW/SnJ, B10.S/SgSf, and B10.PL/SgSf mouse strains which varied in their susceptibility to Coxsackievirus B3-induced immunopathology were suspected to contain autoantibodies against cardiac tissue. These sera were used to identify the target myocardial autoantigen(s). Sera pools were made during the peak of the early, virus-induced myocarditis at 5 and 7 days and during the peak of the late, immunopathic phase of myocarditis at Days 15 and 21 after infection. Only the A.CA/SnJ and A.SW/SnJ strains which develop the immunopathic heart disease had heart-specific autoantibodies as determined by indirect immunofluorescence. This panel of sera pools was then tested against solubilized extracts from whole heart and skeletal muscles. Results from Western immunoblotting analyses demonstrated that antibodies to myosin were a prominent feature in the sera of strains which developed immunopathic myocarditis. The immunopathic sera pools were subsequently assayed against low-salt, high-salt, and a number of detergent extracts of heart and skeletal muscle. Anti-myosin was still the most notable reactivity, even though other autoantigens were detected. Absorption with cardiac myosin removed the vast majority of heart reactivity from the pooled sera derived from the A.CA/SnJ and A.SW/SnJ strains as determined within the limitations of the immunofluorescent and immunochemical assays. Both sarcolemmal and A-band staining patterns were abolished by the cardiac myosin absorption. These studies suggest that myosin is one of the major autoantigens in Coxsackievirus B3-induced autoimmune myocarditis. PMID- 3030578 TI - Calcium, cyclic AMP and protein kinase C--partners in mitogenesis. AB - Evidence is steadily mounting that the proto-oncogenes, whose products organize and start the programs that drive normal eukaryotic cells through their chromosome replication/mitosis cycles, are transiently stimulated by sequential signals from a multi-purpose, receptor-operated mechanism (consisting of internal surges of Ca2+ and bursts of protein kinase C activity resulting from phosphatidylinositol 4,5-bisphosphate breakdown and the opening of membrane Ca2+ channels induced by receptor-associated tyrosine-protein kinase activity) and bursts of cyclic AMP-dependent kinase activity. The bypassing or subversion of the receptor-operated Ca2+/phospholipid breakdown/protein kinase C signalling mechanism is probably the basis of the freeing of cell proliferation from external controls that characterizes all neoplastic transformations. PMID- 3030592 TI - Effect of polybrominated biphenyls on the growth and maturation of human peripheral blood lymphocytes. AB - Polybrominated biphenyls (PBB) have been shown to affect the immune system of exposed Michigan farm workers and their families. The purpose of this study was to evaluate the effects of PBB on the function and on the synthesis of immunoglobulins by peripheral blood lymphocytes. Concentrations of PBB as low as 0.001 microgram/10(5) cells decreased lymphocyte response to pokeweed mitogen; higher concentrations of PBB stimulated the in vitro synthesis and release of immunoglobulins. PBB had no effect on the quantity of E-rosette-forming cells, the total T or B cells, or the ratio of helper to suppressor T-cell subpopulations. Enhanced release of IgG was identified in lymphocyte cultures obtained from blood specimens of PBB-exposed Michigan farmers. The data from this study suggest that PBB exerted an adverse effect on cell function, but produced a nonspecific activation of B lymphocytes. PMID- 3030593 TI - Heritable nail disorders. AB - This article describes a number of syndromes affecting the nail unit. When a patient presents with a complaint involving the nails, it is important to consider first the common nail disorders. However, one should evaluate the entire patient to discover associated signs or symptoms that would point toward an underlying syndrome. The dermatologist may have an opportunity to discover a syndrome that might have significant related morbidity or mortality, which may be prevented through early detection. By recognizing the presence of inherited syndromes, the dermatologist may facilitate genetic counseling. It should be noted that most of the syndromes described are of autosomal dominant inheritance, with variable expressivity, and that the individual manifestations can vary widely in severity, making recognition difficult. PMID- 3030594 TI - Vascular disorders. AB - Cutaneous vascular abnormalities are a feature of many syndromes with multisystemic involvement. The most common associations are hypertrophy of underlying soft tissue and bone, as in the Klippel-Trenaunay-Weber and Sturge Weber syndromes, visceral vascular lesions with hemorrhage, as in hereditary hemorrhagic telangiectasia and blue rubber bleb nevus syndrome, and neurologic alterations, as in Fabry's disease, ataxia-telangiectasia, and the Sturge-Weber syndrome. PMID- 3030597 TI - [Alterations in cholinergic and adrenergic receptors in senile dementia of the Alzheimer's-type]. PMID- 3030595 TI - Pharmacokinetics and acute effect on the renin-angiotensin system of delapril in patients with chronic renal failure. AB - The acute effect on the renin-angiotensin system and the pharmacokinetic properties of delapril, a new angiotensin converting enzyme inhibitor and its active diacid metabolites (delapril diacid and 5-hydroxy delapril diacid) arising from delapril in vivo were investigated in 4 hypertensive patients with chronic renal failure (CRF: 4 males, average age 49.5 (37-64) years, mean Ccr 22.2 ml/min/1.73 m2) and 9 patients with essential hypertension (EH: 6 males, 3 females, average age 42.8 (28-61) years, mean Ccr 79.3 ml/min/1.73 m2). In CRF, following a single dose of delapril hydrochloride (30 mg), the biological half lives (t1/2) of delapril diacid and 5-OH-delapril diacid were 4.69, 12.88 hours, the maximum serum concentration (Cmax) and the area under the plasma concentration-time curve ([AUC]24(0)) of delapril and its diacid metabolites were 414, 797 and 435 ng/ml, and 658, 6400 and 5068 ng X h/ml, respectively. In EH, the t1/2 of delapril diacid and 5-OH-delapril diacid were 1.21, 1.40 hours and the Cmax and [AUC]24(0) of delapril and its diacid metabolites were 489, 635 and 229 ng/ml, and 572, 1859 and 948 ng X h/ml, respectively. The [AUC]24(0) in CRF were significantly increased as compared with those in EH. The cumulative urinary excretions were significantly lower in CRF than in EH. The serum angiotensin converting enzyme (ACE) was markedly inhibited in both groups up to 24 hours. The plasma concentration of angiotensin II decreased in both groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3030598 TI - False positive Meckel's scan caused by carcinoid tumors of the small bowel. AB - A case of gastrointestinal bleeding caused by carcinoid tumor of the small bowel detected by a positive Meckel's scan is presented. This test may prove useful when conventional work-ups fail to localize a bleeding site. PMID- 3030596 TI - Chondroosteogenetic response to crude bone matrix proteins bound to hydroxyapatite. AB - For the development of a new implantable biomaterial that possesses osteogenetic ability, bone matrix morphogenetic proteins were bound to hydroxyapatite (BMP HAP). A crude protein extract including the bone morphogenetic protein (BMP) fraction was precipitated in the pores of hydroxyapatite (HAP) inside of a dialysis sac. In tissue culture, BMP-HAP or HAP was used as substratum for rat mesenchymal muscle cell explants. The BMP-HAP and HAP controls were also implanted in the muscle pouches of mice. New cartilage and/or new bone formation was observed on the exterior surface of BMP-HAP but not of HAP controls. Implantation of this HAP into bone marrow cavities of rabbit femoral and tibial condyle produced new bone formation. The deposits extended further inside BMP-HAP than HAP pores. The pore diameters of 90-200 microns produced earlier ingrowth than into larger pore sizes. BMP-HAP should be tested for clinical application as osteogenetic biomaterial for augmentation of bone regeneration. PMID- 3030599 TI - Technetium-99m RBC scintigraphy in the evaluation of small bowel leiomyoma. AB - Hypervascularity and persistent uptake were demonstrated in a small bowel leiomyoma not actively bleeding at the time of Tc-99m RBC scintigraphy. The potential value of Tc-99m RBC scanning in the evaluation of benign small bowel tumors is discussed. PMID- 3030600 TI - Hernia mimicking testicular torsion. PMID- 3030601 TI - Radionuclide imaging for parotid oncocytoma. PMID- 3030602 TI - Detection of small bowel leiomyosarcoma during a Meckel's scan. PMID- 3030603 TI - Visualization of distal splenorenal shunt by isotopic splenovenography. AB - Radioisotopic angiography of the splenic vein was performed in six patients following a distal splenorenal shunt (Warren procedure). Under echographic guidance, the spleen was punctured with a 22-gauge needle, and Tc-99m pertechnetate was injected into the splenic pulp. In five patients whose esophageal varices had remained atrophic, the splenic vein, the left renal vein, the inferior vena cava, and the heart were clearly visualized within 8 seconds. In one patient, in whom recurrence of esophageal varices had been recognized, the splenic vein was not imaged. The injected material coursed mainly upwards through collaterals. PMID- 3030604 TI - Equivocal findings on cranial CT but apparent cerebral lesion(s) on conventional radionuclide imaging. AB - Four patients with different disease entities (multiple cerebral metastases, cerebral infarct, cerebritis, and encephalitis) in whom x-ray computed tomography was either equivocal or negative showed apparent cerebral lesions by radionuclide studies. Equivocal CTs in the patients with multiple cerebral metastases or cerebral infarction may be attributed to the contraindication of contrast media and/or lack of cooperation during the examination procedure. In patients with cerebritis or encephalitis, radionuclide studies have proven to be more sensitive than CT early in the infectious disease process. In certain circumstances it is clinically beneficial and cost effective to evaluate the patient primarily by radionuclide scintigraphy. PMID- 3030605 TI - Fibrous histiocytoma of the larynx in a child. Case report and review. AB - A 6-year-old child presented with a fibrous histiocytoma of the subglottic larynx. These tumors are rare in adults, and only one other case has been reported in a child. The current case is the youngest and best documented in the pediatric age group. The clinical presentation in this patient, along with controversies related to this class of neoplasms is discussed. PMID- 3030606 TI - Management of neurosurgical problems in pregnancy. AB - The effects of pregnancy on neurosurgical disease are complex and require a detailed knowledge of physiology, anatomy, and pharmacology. The following article details the pathophysiology and treatment of the more common disorders of the nervous system seen during pregnancy. PMID- 3030607 TI - Dynamic exercise-induced increase in lymphocyte beta-2-adrenoceptors: abnormality in essential hypertension and its correction by antihypertensives. AB - We compared the effects of acute stimulation of sympathetic activity by dynamic exercise on a bicycle on lymphocyte beta 2-adrenoceptor density and 10 mumol/L ( )-isoprenaline-evoked lymphocyte cyclic adenosine monophosphate increases in normotensive volunteers with those in patients with essential hypertension. In normotensive subjects exercise increased lymphocyte beta 2-adrenoceptors by about 100%. This effect seems to be a beta 2-dependent process, since it is prevented by propranolol (5 mg administered intravenously) and the beta 2-selective antagonist ICI 118,551 (25 mg t.i.d. orally for 2 weeks) but not by the beta 1 selective antagonist bisoprolol (2.5 mg administered intravenously). In patients with essential hypertension who have elevated lymphocyte beta 2-adrenoceptors, dynamic exercise caused only marginal beta 2-adrenoceptor changes, suggesting an impairment of the acute beta-adrenoceptor regulation. Normalization of blood pressure by antihypertensive treatment resulted in a significant fall in lymphocyte beta 2-adrenoceptors and in a restoration of exercise-induced beta 2 adrenoceptor increases. It is concluded that in essential hypertension the impairment of beta-adrenoceptor regulation is directly linked to the elevated blood pressure. PMID- 3030608 TI - Beta 2-adrenoceptor induced increase of plasma insulin levels in man: evidence of direct and indirect B-cell stimulation and liver effects. AB - Beta 2-Adrenoceptor stimulation is known to increase plasma levels of insulin in man. To study the mechanism behind this increase, plasma insulin, plasma C peptide, and blood glucose responses to the beta 2-adrenoceptor agonist terbutaline were investigated in healthy humans. Terbutaline (125 micrograms i.v.) induced an increase in plasma levels of C-peptide from 0.68 +/- 0.11-0.90 +/- 0.14 nmol/l after 2 min (p less than 0.05) (n = 10). Likewise, plasma insulin concentrations increased, from 0.16 +/- 0.05-0.30 +/- 0.08 nmol/l (p less than 0.02). 20 min after terbutaline injection, plasma levels of C-peptide were 0.98 +/- 0.26 nmol/l and of insulin were 0.21 +/- 0.05 nmol/l. Calculations of the minute to minute increase of plasma levels of C-peptide and insulin during the 20 min after terbutaline injection, revealed that plasma insulin increased during the first 2 min by 59 +/- 8% of the increase of plasma C-peptide, and during the third and fourth min by 44 +/- 12% of the increase of plasma C-peptide; the difference being the liver extraction of insulin. In contrast, during min 4-10 after terbutaline injection, plasma insulin levels increased more than did plasma C-peptide levels, indicating that terbutaline inhibits the liver extraction of insulin. Blood glucose levels did not change during the first 4 min after terbutaline injection. During min 6-10 after terbutaline administration, blood glucose increased, from 4.94 +/- 0.24-5.42 +/- 0.25 mmol/l (p less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3030609 TI - [New prospectives for the treatment of shock: endogenous antagonists of opioids]. PMID- 3030610 TI - The use of electron microscopy and immunohistochemistry in the diagnosis and understanding of lung neoplasms. AB - This article has presented information concerning the ultrastructural features and, to a lesser degree, the immunohistochemical characteristics of lung neoplasms. Electron microscopy and immunohistochemistry are useful techniques for diagnosing and understanding common and rare lung tumors. Electron microscopy is particularly helpful in accurately diagnosing undifferentiated lung tumors such as large-cell undifferentiated and small-cell undifferentiated carcinomas. Ultrastructural studies of lung tumors have also aided in overall understanding of the histogenesis of these tumors. Electron microscopy and immunohistochemistry are techniques that can be applied to tiny specimens, including those obtained via fine-needle aspiration biopsy, and to cells obtained from pleural fluid. An intelligent use of electron microscopy and immunohistochemistry aids in the diagnosis and understanding of lung neoplasms. PMID- 3030611 TI - The ultrastructure of selected gynecologic neoplasms. AB - Several articles have been published recently that discuss the role of electron microscopy in the diagnosis and study of gynecologic neoplasms. It becomes apparent from those works and the review just presented that, although an ultrastructural study is not necessary for reaching a diagnosis of many of these tumors, it may be necessary or supportive in identifying the more poorly differentiated ones. Furthermore, electron microscopy is valuable in providing evidence for the histogenesis of some of these neoplasms. Unfortunately for the pathologist, a certain level of morphologic differentiation (and an absence of metaplasia) in a cell is usually necessary for these goals to be achieved. For example, an adenomatoid tumor (see the article by Dr. Srigley, Mr. Toth, and Mr. Edwards in this issue) of the fallopian tube can readily be accepted as being composed of mesothelial cells, because both the neoplastic cells and normal mesothelial cells have the same highly differentiated features of long, slender microvilli, prominent intercellular junctions, and many microfilaments. On the other hand, there is very little resemblance between the granulosa cells of a granulosa-cell tumor and mature mesothelial cells. Thus, if one of the theories of histogenesis of granulosa cells were correct--namely, that they are derived ultimately from mesothelial lining--the ultrastructural evidence would rest on recognizing a similarity between the two types of cells at an earlier stage of differentiation. The neoplastic granulosa cell has differentiated along a separate, specialized line in which the ultrastructural resemblance to the parent cell is partly, if not completely, lost. Another example of the type of information that electron microscopy can provide is in relation to the common epithelial tumors. There is good evidence that the serous tumors in this group arise from mesothelium, although ultrastructurally their differentiation has veered from a mesothelial direction to one in which the cells have a complement of organelles related to secretory activity. Paradoxically, the mucinous cystic tumors, which have been classified traditionally as tumors of surface epithelial origin, are now thought to be of germ-cell origin in some cases, as examples of monophyletic teratomas. The ultrastructural evidence for this conclusion rests on the presence of anchoring filaments in the microvilli of the neoplastic cells, similar to those of normal intestinal epithelium, and on an admixture of various types of gastrointestinal cells, including those that contain dense-core granules (argentaffin cells).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3030613 TI - Tumors of the liver, extrahepatic biliary tract, and pancreas. AB - Electron microscopy can be of limited value in distinguishing among poorly differentiated hepatocellular carcinomas, bile duct carcinomas, and other adenocarcinomas, but may be very helpful in identifying tumors that are metastatic to the liver. The characteristic ultrastructural features of endocrine and exocrine tumors of the pancreas are readily distinguished by electron microscopy when the tissue is adequately preserved, and this technique can be very useful in establishing a diagnosis. PMID- 3030612 TI - Central nervous system tumors. AB - In this article the authors deal with the morphology of primary tumors of the central nervous system and its coverings. The discussion includes astrocytoma variants/glioblastoma, ependymoma and its variants, subependymoma, choroid plexus papilloma and carcinoma, embryonal CNS tumors, neuronal neoplasms, meningioma, dural hemangiopericytoma (angioblastic mengioma), and hemangioblastoma. PMID- 3030614 TI - Small-cell pediatric tumors: histology, immunohistochemistry, and electron microscopy. AB - Histology, immunohistochemistry, and electron microscopy give an account on what is known of the correlation between histologic appearance and prognosis in this important group of tumors. Ultrastructural and immunohistochemical findings are presented that frequently are useful in establishing a diagnosis, but also serve to elucidate the histogenesis of some tumors, especially those that lack characteristic features. PMID- 3030615 TI - Diagnostic electron microscopy of male genital tract tumors. AB - As illustrated in this review, neoplasms of the male genital tract are markedly heterogenous, reflecting their complex embryologic derivation and histogenetic classification. Transmission electron microscopy has greatly increased our understanding of the structure of these tumors, and in doing so has greatly improved our light microscopy. Additionally, in a number of selected situations, TEM also provides important practical diagnostic information. Currently, light microscopy in conjunction with clinical information is the central tool of tumor taxonomy. However, TEM with histochemistry, immunohistology, analytic cytometry, and molecular biology provide practical and useful information in some situations. In all cases, the application of these techniques has greatly increased our overall understanding of tumor structure, pathobiology, and classification. PMID- 3030616 TI - Long-term high fibre, low fat diet in gestational diabetes. AB - A 30-year-old Caucasian who developed gestational diabetes in her first pregnancy requiring 58 U insulin daily and who subsequently adopted a high fibre, low fat diet and who was able to maintain normal glucose tolerance throughout a second pregnancy is reported. PMID- 3030617 TI - Guar: pharmacological fibre or food fibre? PMID- 3030618 TI - Acceptability of high-fibre diets in diabetic patients. AB - Twenty-five diabetic patients were selected from Oxford Diabetic Clinics to assess their dietary compliance (Group A). All had been taught to follow diets rich in dietary fibre in which carbohydrate provided 50% of total energy. The results were compared with those obtained previously from a similar group of patients (Group B) all of whom had been instructed to follow a diet in which carbohydrate provided 40% of total energy. The patients of Group A had a significantly greater intake of carbohydrate (45.1% vs 34.7% of total energy) and dietary fibre (33.1 g vs 18.7 g) and a lower intake of fat (33.5% vs 42.1% of total energy) than the patients of Group B. The results of dietary assessment obtained from a third group of patients (Group C), who had been advised to follow a high-fibre diet before the widespread availability of dietary teaching aids and recipe books, showed that carbohydrate and fat provided 37.5% and 41.0% of total energy, with a dietary fibre intake of 25.6 g. The results suggest that patients are willing and able to change their dietary habits towards a distribution of food constituents likely to improve diabetic control and reduce the risk of coronary heart disease when given enthusiastic instruction and support in diabetic clinics. PMID- 3030619 TI - Diabetic control is improved by guar gum and wheat bran supplementation. AB - Twenty-eight insulin-dependent diabetics were treated with different dietary regimes for three periods of three months. Initially they used a white flour bread (run-in period), then their daily bread ration was enriched with guar gum (mean dose: 29 g), and then with wheat bran (mean dose: 33 g) in a randomized crossover pattern. Fasting and postprandial blood glucose levels were measured on filter paper spots collected once weekly at home, and other biochemical values were measured monthly. No improvement in diabetic control was seen during the run in period. Mean postprandial blood glucose decreased from 12.0 +/- 3.8 mmol/l (mean +/- SD) in the run-in period to 9.7 +/- 2.8 mmol/l (p less than 0.01) in the guar period and to 9.7 +/- mmol/l (p less than 0.01) in the bran period. HbA1 decreased from 10.5 +/- 2.1% in the run-in period to 9.7 +/- 1.6% (less than 0.05) at the end of the guar period and 9.9 +/- 1.2% (not significant) at the end of the bran period. Only modest changes were seen in serum-lipids--total cholesterol decreased significantly in the guar period, but not in the bran period. In this study both guar gum and wheat bran were well tolerated and produced a substantial decrease in postprandial blood glucose. PMID- 3030620 TI - Influence of metabolic control of insulin-dependent diabetes on plasma nucleotide levels (cAMP, cGMP) during bicycle exercise. AB - Plasma concentrations of cyclic nucleotides (cAMP, cGMP) were measured before and during bicycle exercise in 8 well-controlled (mean pre-exercise blood glucose 5.3 mmol/l; HbA1 8.6%) and 8 moderately controlled (mean pre-exercise blood glucose 12.2 mmol/l; HbA1 10.8%) patients aged 18-32 years with insulin-dependent diabetes mellitus (IDDM) and in a group of non-diabetic control subjects matched for age and sex. Pre-exercise plasma cAMP concentrations and the rise with exercise were similar in all study groups. Significantly lower resting cGMP levels were found in well-controlled IDDM patients (3.5 +/- 0.3 pmol/ml, mean +/- SEM) compared to controls (5.6 +/- 1.1 pmol/ml; p less than 0.05) and moderately controlled IDDM patients (5.6 +/- 1.0 pmol/ml; p less than 0.05). By contrast, plasma cGMP levels increased during exercise in the diabetics but not in the controls. These findings indicate a significant difference in responses of plasma cGMP to exercise between IDDM patients and controls. PMID- 3030621 TI - Angiotensin-converting enzyme (ACE): relationship to insulin-dependent diabetes and microangiopathy. AB - Angiotensin-converting enzyme (ACE) is secreted by the vascular endothelium and serum activity may reflect endothelial damage. A study of 48 insulin-dependent diabetics, 15 with and 33 without evidence of diabetic retinopathy and 41 non diabetic controls was performed. ACE activity was significantly elevated in the diabetics compared with controls (mean +/- SD 46 +/- 14 vs 35 +/- 9 U/l, p less than 0.001) (units in micromoles substrate converted/min/l serum). This elevation was more marked in diabetics with such evidence of microangiopathy as retinopathy or raised albumin excretion rate (AER) (51 +/- 14 U/l, p less than 0.0001), and also in those with raised AER alone (47.2 +/- 15 U/l, p less than 0.002). Patients with both raised AER and retinopathy had significantly higher ACE activities than those with no complications (53 +/- 15 vs 41.2 +/- 15 U/l, p less than 0.05). No correlation was found with glycosylated haemoglobin or smoking habits. We conclude that mean serum angiotensin converting enzyme activity is increased in insulin-dependent diabetes, particularly in those with evidence of microangiopathy and this may reflect microvascular damage. PMID- 3030622 TI - Guar gum: the importance of reporting data on its physico-chemical properties. PMID- 3030623 TI - Lawrence Lecture. Hypoglycaemia and diabetes. PMID- 3030624 TI - Ambulatory monitoring of the ST segment in diabetic men with and without peripheral neuropathy. AB - To assess whether myocardial ischaemia is more common in diabetic patients with neuropathy, 24-hour ambulatory monitoring of the ST segment was performed on 27 diabetic men without peripheral neuropathy and in 28 with neuropathy. The patients were matched for age 54 +/- 7 years (mean +/- SD) versus 54 +/- 7 years and for duration of diabetes (16 +/- 9 years versus 16 +/- 12 years). None had clinical evidence of heart disease. Episodes of ST segment depression were seen during ambulatory monitoring in 12 diabetics (22%) but were not more common in patients with peripheral neuropathy. Four of the 13 diabetics with autonomic neuropathy had ST depression during ambulatory monitoring. During a median follow up period of 50 months, four patients developed clinical heart disease. Three of these patients had shown ST depression during ambulatory monitoring. ST depression during ambulatory monitoring is common in diabetic men without cardiac symptoms but is not related to the presence of peripheral neuropathy. Diabetics with ST depression during ambulatory monitoring are at increased risk of developing heart disease in subsequent years. PMID- 3030625 TI - Activities of adenylate-degrading enzymes in muscles from vertebrates and invertebrates. AB - Activities of adenylate-degrading enzymes in muscles of vertebrates and invertebrates were determined. Mammalian and fish muscles showed a markedly higher activity of AMP deaminase with a lower level of adenosine deaminase and 5' nucleotidase. Cephalopods showed an active adenosine deaminase and a 5' nucleotidase which preferred AMP as the substrate. Negligible deamination of AMP and adenosine and little phosphohydrolase activity toward AMP and IMP were observed in the shellfish muscles. Adenine nucleotides can be degraded to form IMP via the AMP deaminase reaction in vertebrate muscles, while dephosphorylation of AMP to adenosine, which is then converted to inosine, appears to proceed in cephalopods. Adenylates can be hardly degraded in shellfish muscles. PMID- 3030626 TI - Levels of glycerate 2,3-P2, 2,3-bisphosphoglycerate synthase and 2,3 bisphosphoglycerate phosphatase activities in rat tissues. A method to quantify blood contamination of tissue extracts. AB - The levels of glycerate 2,3-P2 and of 2,3-bisphosphoglycerate synthase and 2,3 bisphosphoglycerate phosphatase activities have been determined in isolated rat hepatocytes and adipocytes and in perfused rat tissues to discard blood contamination. The values obtained are much lower than those previously reported, ranging 0.50-40 nmol/g tissue. No relationship appears to exist between glycerate 2,3-P2 concentration and the levels of the enzymatic activities involved in glycerate 2,3-P2 metabolism. Assay of glycerate 2,3-P2 in tissue extracts constitute a very useful way to quantify blood contamination. PMID- 3030627 TI - Protein methylase II in five taxa from three phyla. AB - Protein methylase II (protein O-methyltransferase, EC 2.1.1.24) was found in Dictyostelium discoideum amoebae, Astacus leptodactylus axonal, Locusta migratoroides neuronal, Torpedo marmorata electroplaque and Bos bovis stratial tissue and compared in both the soluble and particulate fractions. The intrinsic decay data of the methyl groups transferred onto proteins from Dictyostelium and Torpedo tissues were virtually identical. The short term kinetics of the methyl group transfer of all fractions and of all taxa investigated were non-linear and multiphasic. The particulate fractions displayed transient peaks at 1 min, 3 min, or both after the start of the reaction. The methyl group transfer was stimulated by the neurotoxins veratridine (VTx) and inhibited by veratridine plus tetrodotoxin (TTx) (axonal membrane vesicles of Astacus), stimulated transiently and in a biphasic manner by carbamoylcholine and phospholipase A2 (AChR-rich membrane vesicles of Torpedo), and stimulated transiently and biphasically by the adequate chemotactic stimulus cAMP (aggregation competent amoebae of Dictyostelium). PMID- 3030629 TI - Breast carcinoma metastatic to the palatine tonsils. PMID- 3030628 TI - Cytosol 5'-nucleotidase from Artemia embryos. Purification and properties. AB - Cytosol 5'-nucleotidase (EC 3.1.3.5) has been purified near homogeneity from Artemia embryos. The enzyme cleaves preferentially IMP and GMP, and to a lesser extent other 5'-mononucleotides. The substrate-velocity plot was hyperbolic with GMP and sigmoidal with AMP. The hydrolysis of GMP is stimulated both by ATP and beta, gamma-methyleneadenosine 5'-triphosphate with the same activation constant of around 0.6 mM. Both nucleotides decreased S0.5 without affecting V. The molecular mass of the native purified enzyme was 165 kDa, and one major band of 42 kDa was detected after sodium dodecyl sulphate polyacrylamide gel electrophoresis. PMID- 3030630 TI - Inhibition by gossypol of cyclic AMP production in mouse Leydig cells. AB - Gossypol has been shown to inhibit steroidogenesis in leydig cells. This study examined the mechanism of this action by investigating the effect of gossypol on leydig cell hCG receptor binding, adenylate cyclase activity and cyclic AMP production in leydig cells. Gossypol had no effect on hCG binding to cell membranes but inhibited LH-stimulated cyclic AMP production in whole purified leydig cells. It also reduced the stimulation caused by LH, Gpp(NH)p, forskolin and fluoride in membranes from leydig cells and also from liver. It is therefore possible that gossypol affects cyclic AMP production at the level of ATP conversion to cyclic AMP, although effects on the G-protein and catalytic subunit cannot be ruled out. PMID- 3030631 TI - Recent advances in Bacteroides genetics. AB - Bacteroides are Gram-negative, obligate anaerobes that are present in high concentrations within the intestinal tracts of humans and animals. Bacteroides are also important opportunistic pathogens of humans and animals. Methods for genetic manipulation of these important organisms have only recently begun to emerge. Shuttle vectors which can be transferred by conjugation between Escherichia coli to Bacteroides are now available. A method for transforming some strains of Bacteroides has been developed. Two Bacteroides transposons, Tn4351 and Tn4400, have been found and one of them, Tn4351, has been used for transposon mutagenesis of Bacteroides. Several different Bacteroides genes have now been cloned, including a gene that codes for resistance to clindamycin, genes that code for polysaccharidases (chondroitin lyase and pullulanase), and a gene that codes for a fimbrial subunit. These cloned genes have been used to study the organization and regulation of Bacteroides genes. PMID- 3030633 TI - Peripheral adrenergic stimulation and indomethacin in experimental ocular shedding of HSV. AB - The application of 6-hydroxydopamine to the cornea by iontophoresis, followed by topical epinephrine, effectively induces herpes simplex virus (HSV) shedding from the external eye of latently infected rabbits. In this study the beta adrenergic blocker, Timolol, reduced virus shedding when applied immediately before the epinephrine, but continued administration resulted in increased viral shedding. While indomethacin, a prostaglandin synthesis inhibitor decreased HSV replication in cell culture, it failed to decrease virus shedding when applied topically to the eye in adrenergically stimulated animals. Timolol may act then by its effect on the peripheral cells of the eye rather than by stimulation of virus production in ganglionic neurons. These same animals were subsequently tested for latent infection of the trigeminal and superior cervical ganglia and corneas 14 months after primary infection. Only 2 of 14 animals had virus in the trigeminal ganglia, a finding which suggests that latent virus may be depleted by repeated reactivations. Virus was recovered from corneas of five rabbits by co-cultivation so it is possible that corneal latency occurs in this rabbit model as it does in humans. PMID- 3030632 TI - Ocular pathogenicity of herpes simplex virus. AB - As a relatively small, discrete organ that contains a number of widely different cell types the eye provides an intriguing system in which to study fundamental aspects of virus/cell interactions. Such aspects are considered with particular reference to herpes simplex virus and the pivotal role of virus/neuron interactions in the development of ocular disease. Three aspects of this interaction are discussed: the entry of virus into the eye latency in the trigeminal ganglion nerve damage. PMID- 3030634 TI - Identifying HSV infected neurons after ocular inoculation. AB - ICR mice were inoculated intracamerally with McKrae strain herpes simplex virus (HSV) followed by intraperitoneal injection with 3H-thymidine. Infected mice were sacrificed after 3 or 4 days and the eyes, trigeminal ganglia (TG) and superior cervical ganglia (SCG) were embedded in glycol methacrylate, sectioned, and dipped for autoradiography. Light microscopy revealed silver grain labeling over neurons in the ipsilateral retina, TG and SCG of infected animals. No labeling of neurons was noted in the contralateral TG or SCG. Since the DNA of mature neurons does not replicate, we interpret these labeled neurons to represent cells with active replication of HSV. This technique allows the study of HSV infection of the nervous system with excellent tissue preservation. Furthermore, it may be used to distinguish those neurons with intrinsic viral synthesis from those harboring virus synthesized at a distant site with subsequent intracellular spread. PMID- 3030635 TI - A critical role for ACAID in the distinctive pattern of retinitis that follows anterior chamber inoculation of HSV-1. AB - To test the hypothesis that ACAID induction is instrumental in producing the distinctive pattern of retinal pathology that follows anterior chamber inoculation of HSV-1 in BALB/c mice, panels of mice received uniocular anterior chamber, uniocular intravitreal, and bilateral anterior chamber inoculations of HSV-1. It was found that contralateral retinitis developed after the first two routes, and ACAID was induced by all three. Enucleation of eyes inoculated with HSV-1 before 3 days post-inoculation (but not thereafter) prevented both ACAID and contralateral retinitis. Intracameral inoculations of HSV-2 induced vigorous delayed hypersensitivity and failed to incite contralateral retinitis. It is concluded that ACAID induction plays a crucial role in the pathogenesis of contralateral retinitis following anterior chamber inoculation of HSV-1. PMID- 3030636 TI - Virus-specific DTH prevents contralateral retinitis following intracameral inoculation of HSV-2. AB - Following inoculation of HSV-1 (KOS strain) into the anterior chamber of one eye of a BALB/c mouse, the virus travels to the uninoculated contralateral eye and contributes to the devastating retinal necrosis which occurs in this eye 10-12 days p.i. In parallel experiments, HSV-2 (MS strain) was inoculated uniocularly via the anterior chamber route; animals were sacrificed at intervals and their eyes removed for histopathology and virus culture. Histopathological examination revealed that, in contrast to the retinal destruction observed in HSV-1 inoculated animals, the retina of the contralateral eye of HSV-2 injected animals was unaffected. Culture studies demonstrated that, similar to the spread of HSV-1 after anterior chamber inoculation, HSV-2 reached the contralateral eye in two temporally separate waves and also revealed that the amount of virus recovered from the uninoculated eye of HSV-2 infected animals was significantly less than that recovered from the uninoculated contralateral eye of HSV-1 injected mice. Further examination demonstrated that animals inoculated with HSV-2 via the anterior chamber route did not develop the suppression of virus-specific delayed hypersensitivity (DH) which is characteristic of HSV-1 induced anterior chamber associated immune deviation (ACAID). Taken together, the sparing of the contralateral retina and the ability to generate a virus-specific DH response suggest that DH assists in the preservation of the contralateral retina of animals inoculated with HSV-2 via the anterior chamber route by limiting virus replication and the amount of virus which reaches the uninoculated eye. PMID- 3030638 TI - HSV-induced blastogenesis in splenic mononuclear cells from inbred mice. AB - The severity of HSV stromal keratitis varies among inbred mouse strains, with A/J and BALB/c greater than C57BL/6. Since cell-mediated immunity (CMI) is thought to play a role in the pathogenesis of HSV stromal keratitis, we measured the proliferative response of primed splenic mononuclear cells (SMC) from these strains to HSV antigen in vitro. Primed SMC from all three strains showed increased thymidine uptake after incubation with UV-inactivated HSV-1 antigen (UV HSV) in vitro when compared with control antigen. Uptake by SMC from BALB/c and A/J mice was greater than that by SMC from C57BL/6 mice. The difference between BALB/c and A/J was not significant. Uptake by non-primed SMC cultured with UV-HSV was not significantly greater than uptake induced by control antigen. Thus, among the strains studied, A/J and BALB/c mice, which are relatively susceptible to stromal keratitis, have the greatest proliferative response to UV-HSV antigen. C57BL/6 mice, which are relatively resistant to stromal keratitis, have the least response. These findings suggest that genetically determined differences in CMI may influence the course of HSV keratitis and are consistent with the hypothesis that the host immune response plays a role in the pathogenesis of HSV stromal keratitis. PMID- 3030637 TI - Class II induction of human corneal fibroblasts by cell-free supernatants from HSV stimulated lymphocytes. AB - Lymphocytes obtained from patients with herpes simplex virus, type 1 (HSV) neutralizing antibodies were stimulated in vitro for 48 hr with heat inactivated HSV. Cell-free supernatant from these cultures was added to autologous or allogeneic HLA-DR-negative stromal (fibroblast) cells for 3 days. All stromal cultures were stained with monoclonal reagents by indirect immunofluorescence for Class I (beta 2) or Class II (HLA-DR) antigen expression. Supernatant from HSV stimulated lymphocytes induced HLA-DR antigen on cultured stromal cells while supernatant from lymphocytes cultured in the absence of HSV antigen or from lymphocytes which did not proliferate in response to incubation with HSV antigen did not induce HLA-DR. The expression of Class II antigens induced by supernatant from HSV-stimulated lymphocytes was inhibited by the addition of purified antibodies to IFN. PMID- 3030639 TI - HSV-1 thymidine kinase negative vaccine: pathogenicity, protection, and perils. AB - Primary inoculation of mice and rabbits with an avirulent HSV-1 thymidine kinase negative (TK-) mutant reduced keratitis, mortality, and superinfection of the trigeminal ganglion (TG) as measured by cocultivation and iontophoresis-induced ocular shedding following ocular challenge with HSV-1 McKrae and W strains. However species differences were demonstrated; with complete protection in rabbits, and incomplete protection in mice. In mice, BUDR/autoradiography and restriction enzyme analysis identified both HSV-1 McKrae and W strains as having superinfected the TG, established latency and reactivated. Also, the avirulent TK negative inoculating strain was altered to a virulent TK positive strain through possible in vivo selection, mutation &/or host-modification, and possible in vivo recombination with the virulent challenge strains. We conclude that lower species differences require that potential HSV-1 vaccines be tested in non-human primates prior to clinical trials, and that a DNA-free subunit herpes vaccine represents a safer alternative to a live virus vaccine. PMID- 3030640 TI - Protective effect of passive immunization on herpetic retinitis of newborn rabbits. AB - We studied the protective effects of passive immunization with virus specific antibody in newborn rabbits inoculated subcutaneously with type 2 herpes simplex virus (HSV-2). Newborn rabbits given anti-HSV-2 antibody intraperitoneally (IP) on days 0, 2 and 4 post infection had smaller herpetic skin lesions and reduced mortality when compared to controls. In addition, the IP treatment using this schedule reduced virus growth in the skin lesions and virus dissemination, so that it decreased the frequency of herpetic retinitis. When the IP antibody administration was started at 24 hours post virus inoculation, according to the schedule days 1, 3 and 5, there was less protection; larger skin lesions, higher mortality, and greater evidence of virus dissemination. Also HSV-infected mononuclear cells (MNCs) treated with anti-HSV serum resulted in a significant reduction in the number of infected MNCs. The results of these studies suggest that anti-HSV-2 antibody contributes to protection against HSV-2 infection of skin as well as eyes, probably by inactivation of the virus locally at the skin inoculation site, and by combating the hematogenous spread of HSV-infected MNCs as well as free virus to various organs including the eye. PMID- 3030641 TI - Protective immunity against herpetic ocular disease in an outbred mouse model. AB - Immunization of outbred mice by subdermal (footpad) inoculation with the F strain of herpes simplex virus type 1 (HSV-1) induces an immune response which protects the animals against herpetic ocular disease and encephalitis, and reduces the incidence of latent trigeminal ganglion infections following corneal challenge with the RE strain of HSV type 1. The protective effects are proportional to the dose of virus used for immunization. Heat-killed virus preparations also protected the mice against encephalitis and stromal keratitis, but failed to prevent epithelial keratitis and establishment of latency. PMID- 3030642 TI - Passive immunization with monoclonal antibodies against herpes simplex virus glycoproteins protects mice against herpetic ocular disease. AB - The effects of passive immunization with specific monoclonal antibodies against herpes simplex virus glycoproteins gB, gC, gD, and gE on the course of herpetic keratitis, survival and the establishment of latency in an outbred mouse model are described. A total of nine monoclonal antibodies were tested in these experiments. Passive immunization at 24 or 48 hours post-inoculation had little effect on the severity of the initial epithelial infection of the cornea, but blocked dissemination of the virus to the central nervous system and periocular tissues and prevented development of blepharitis, iritis and stromal keratitis. Additional studies are needed to characterize these monoclonal antibodies in greater detail, and to define the mechanism of these protective effects. PMID- 3030643 TI - Nucleotide sequences important in DNA replication are responsible for differences in the capacity of two herpes simplex virus strains to spread from cornea to central nervous system. AB - Two herpes simplex virus (HSV) intertypic recombinants were isolated with genomes composed entirely of HSV-2(186) nucleotide sequences except for a 6.0 kb segment of HSV-1(17) DNA positioned between 0.40 and 0.44 map units. Following corneal infection of mice, HSV-1(17) and the two intertypic recombinants spread from infected eyes into the central nervous system and induced a fatal encephalitis. Ocular infection with the HSV-2(186) parent did not lead to detectable amounts of virus in the brain, and none of the mice developed encephalitis. The 6.0 kb HSV 1(17) DNA inserted within the genome of the two intertypic recombinants contained nucleotide sequences involved in DNA replication. These include the HSV-1(17) oriL, the HSV-1(17) gene for DNA polymerase and portions of the HSV-1(17) gene coding for DNA-binding protein ICP8. Thus, our results indicate that the difference in the capacity of HSV-1(17) and HSV-2(186) to spread from the cornea into the CNS is determined solely by nucleotide sequences associated with DNA replication. PMID- 3030644 TI - Antiviral sensitivities of the acute retinal necrosis syndrome virus. AB - Varicella zoster was isolated from the vitreous of a patient with the acute retinal necrosis (ARN) syndrome. We utilized a plaque reduction assay to determine the in vitro susceptibility of the ARN isolate to 6 antiviral drugs. The effective doses for 50% inhibition of plaque numbers were 5.3 microM for for acyclovir, 4.7 microM for DHPG, 8.7 microM for ARA-A, 100.7 microM for phosphonoacetic acid, 0.07 microM for BVdU and 2.4 microM for IUdR. Similar inhibitory values were obtained for the OKA vaccine strain of varicella zoster virus. These data do not support the notion that the ARN strain may represent a mutant of varicella zoster virus with significant alterations in either the viral thymidase kinase or DNA polymerase genes based upon its antiviral sensitivities. The implications of these results regarding the role of antiviral chemotherapy in the ARN syndrome are discussed. PMID- 3030645 TI - Anti-herpes simplex virus (HSV) effect of 9-(1,3-dihydroxy-2 propoxymethyl)guanine (DHPG) in rabbit cornea. AB - The anti-herpes simplex virus (HSV) effect and cytotoxicity of a new nucleoside analogue, 9-(1,3-dihydroxy-2-propoxymethyl)guanine (DHPG) in rabbit cornea were studied. In tests of the anti-HSV effect of DHPG, even 0.03% ointment, given 5 times per day for 2 days, prevented lesion formation. The preventive effect of DHPG was much stronger than that of acyclovir (ACV) or 5-iodo-2'-deoxyuridine (IDU). In tests on the therapeutic effect of DHPG against dendritic ulcers, 0.3% ointment, given 5 times per day for 4 days, had a dramatic therapeutic effect. The effect was stronger than that of 3% ACV or 0.5% IDU ointment. Application of 0.3%, 1% or 3% DHPG ointment to normal rabbit corneas, 5 times per day for 2 weeks resulted in no histopathological abnormalities. The above results show that DHPG is superior to ACV or IDU for treatment of HSV infections. PMID- 3030646 TI - 2'-nor-cGMP, a new cyclic derivative of 2'NDG, inhibits HSV-1 replication in vitro and in the mouse keratitis model. AB - The present study examined the antiherpetic effect of 2'-nor-cGMP, a new cyclic phosphate derivative of 2'NDG, in vitro and in the mouse keratitis model. The 50% inhibitory dose (ID50) was determined with HSV-1 RE strain in Vero cell monolayers for 2'-nor-cGMP (6.9 mcg./ml), 2'NDG (.06 mcg/ml), and trifluridine (F3T) (.72 mcg/ml). Balb C mice underwent bilateral ocular inoculation with HSV-1 RE strain, and then were treated with different therapeutic regimens. The antiviral efficacy of each drug was evaluated by ocular virus titers, clinical grading of epithelial keratitis, and histological evaluation of stromal keratitis. 2'-nor-cGMP was the most effective drug (P = .0001) in reducing ocular viral titers. Both 2'-nor-cGMP and 2'NDG were significantly more effective (P = .0001) than F3T in reducing epithelial keratitis, and as effective as F3T in reducing stromal keratitis. PMID- 3030647 TI - The antiherpesvirus activity and cytotoxicity of sangivamycin. AB - Sangivamycin, 4-amino-5-carboxamido-7-(beta-D-ribofuranosyl)-pyrrolo[2,3-d] pyrimidine is a structural analog of adenosine belonging to a group of nucleosides classified as pyrrolopyrimidines. Sangivamycin, an adenosine deaminase resistant analog, was found to inhibit the replication of three strains of herpes simplex virus type 1 (HSV-1) by 50% (ED50) at a concentration approximately equal to the concentration which inhibits cell growth by 50% (LD50). Both Vero cells and rabbit corneal stromal cells in exponential growth were about 10-fold more sensitive to the drug than quiescent cells. The selectivity indices of sangivamycin indicated that the drug was not a highly selective antiviral agent and, therefore, would offer no advantage over drugs currently available for the treatment of herpetic keratitis. PMID- 3030648 TI - Reevaluation of human alpha interferons against herpesvirus infection of the rabbit eye. AB - Because of reported differences in potency, recombinant DNA-derived human alpha interferons (IFNs) were reevaluated for use against acute Herpes Simplex Virus type 1 (HSV-1) infection of the rabbit eye. The IFNs used topically were IFN alpha 2, (IFN-alpha 2) and a consensus of known human IFN-alpha s, designated IFN alpha Con1. Prophylactic treatment with IFN-alpha Con1 at 1 or 15 X 10(6) U/eye/day beginning 48 hours before HSV-1 inoculation and therapeutic treatment with 5 or 15 X 10(6) U/eye/day beginning 4 hours after inoculation with either IFN-alpha Con1 or IFN-alpha 2 appeared to prevent or significantly reduce the development of corneal epithelial involvement. The effects were dose dependent with no evidence for decreased activity at the higher dose. The duration of HSV-1 shedding into tear film was not significantly reduced. PMID- 3030649 TI - Failure of intertypic recombinant constructed from HSV-1 x HSV-2 virulent parents to induce ocular pathology. AB - An intertypic recombinant constructed from HSV-1 x HSV-2 parents was isolated which failed to induce any overt ocular pathology when inoculated onto the sacrificed cornea of four-week-old SJL/J mice. During the 24-48 hour post infection period there was transient virus replication but by day 3 the infectious titer in the eye had dropped by greater than or equal to 10(4)-fold, and little or no virus could be recovered thereafter. When immunosuppressed (600 r) mice were infected corneally, virus clearance was delayed several days but again no obvious ocular pathology was seen, and no mice died. By contrast, infection of the cornea with either parent was followed by virus replication and development of clinically apparent pathology which could progress to blinding stromal keratitis. The genome of the intertypic recombinant was analyzed by agarose gel electrophoresis of restriction endonuclease digests and found to consist entirely of HSV-1 DNA except for HSV-2 DNA sequences located between map units 0.10-0.16, 0.41-0.43, and 0.77-1.0. Potential explanations for the loss of virulence are discussed. PMID- 3030651 TI - Clinical findings after zosteriform spread of herpes simplex virus to the eye of the mouse. AB - When herpes simplex virus (HSV) is inoculated onto the snout of the inbred strain NIH mouse, clinical disease of the eye ensues only after a delay, due to spread of virus to the eye occurring via neural pathways. This report is concerned with the detailed description of eye disease. Physical signs observed include mydriasis, iritis and keratitis. The incidence of combinations of physical signs has been analysed by the computer and presented as pie-charts to show the complexity and evolution of the eye disease. PMID- 3030650 TI - The potency of interferon-alpha 2 and interferon-gamma in a combination therapy of dendritic keratitis. A controlled clinical study. AB - Forty-five patients with virologically confirmed dendritic keratitis were treated in a randomized, double-blind controlled study with a basic therapy of trifluorothymidine (TFT) eye drops. In addition they received different human recombinant interferon (rHu IFN) eye drops. The following results were obtained for average healing times: TFT plus one drop daily of rHu IFN-alpha 2 arg (30 million iu/ml): 3.3 days, TFT plus rHu IFN-gamma (30 million iu/ml): 3.9 days, TFT plus a mixture of alpha plus gamma (0.3 million iu/ml each): 6.1 days, TFT plus a mixture of alpha plus gamma (1.5 million iu/ml each): 3.3 days. High-titer gamma interferon did not significantly differ from high-titer alpha interferon in the combination therapy of dendritic keratitis. A mixture of alpha plus gamma at a moderate titer (1.5 million iu/ml each) was as effective as a high-titer mono preparation. Adding a low-titer interferon mixture gave no better therapeutic results than antiviral monotherapy. Thus it seems possible to save about 90% of interferon commonly used in the combination therapy of dendritic keratitis by applying a mixture of different suitable interferons instead of interferon monospecies. PMID- 3030652 TI - Analysis of glycoproteins expressed by isolates of herpes simplex virus causing different forms of keratitis in man. AB - An in vitro analysis of glycoprotein produced by nine human ocular isolates of HSV-1 is reported. The source of the isolates was; three patients with recurrent dendritic keratitis, three with chronic stromal disease and three with primary keratoconjunctivitis. Virus strains were labelled with the radioactive precursors (35S) methionine and (14C) glucosamine. Radiolabelled viral glycoproteins were subsequently analysed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), followed by autoradiography. Viral glycoproteins were further characterised by immuno-precipitation with polyclonal and monoclonal antibodies to HSV. The stromal isolates excrete larger amounts of 'soluble' precursor glycoprotein D than those in the other two disease categories. It is possible that the immune response to glycoprotein D is in part responsible for the severity of stromal disease. PMID- 3030653 TI - HSV1 strain sensitivity in experimental rabbit keratitis: evolution under repeated topical IDU administrations. AB - The effects of repeated topical idoxuridine (IDU) administration of HSV1 strain sensitivity were investigated during 6 serial passages (P1 to P6) in the rabbit. By comparison to placebo treated rabbits, a delay in ulcer cicatrization appeared at P2 and clinical resistance was completed at P3. Clinical cross resistance to acyclovir (ACV) was also tested and demonstrated at P7. In vitro, a plaque reduction test on Vero cells using directly the tear film HSV populations allowed the prediction of the resistance by an early rise in the effective dose 90% (ED 90) value anticipating that in ED 50%. An ED 50 determination by dye-uptake assay on P6 HSV isolate demonstrated a cross resistance to viral thymidine kinase (TK) dependent drugs without any change in Ara-A and PFA sensitivity, according to a 23% TK activity at P6. At the last passage the HSV drug resistant population had an unrestricted corneal pathogenicity. A return to IDU and ACV in vitro sensitivity was demonstrated in group control animals at P2 but not at P4 or P6. PMID- 3030654 TI - Routes of viral spread in the von Szily model of herpes simplex virus retinopathy. AB - Intraocular inoculation of herpes simplex virus type 1 (HSV-1) in one eye of rabbits results in encephalitis and contralateral necrotizing viral retinopathy. The effects of viral inoculation site and optic nerve (ON) transection on the spread of virus to the brain and contralateral eye in this model were investigated. A surgical technique was developed for transection of the retrobulbar optic nerve posterior to the entrance of the central retinal vessels. HSV-1 was inoculated into the AC or vitreous of one eye in normal rabbits and in rabbits with one ON transected, either ipsilateral or contralateral to the side of inoculation. Animals were followed clinically for signs of disease. Encephalitis and contralateral retinopathy (CR) occurred following both AC and vitreous inoculation of virus, although CR developed later in AC-inoculated rabbits. Ipsilateral retinopathy (IR) developed in 83% of vitreous-inoculated rabbits, but in only 5% of AC-inoculated animals. IR developed 8 days after the onset of CR in the AC-inoculated group. ON transection on the side of virus inoculation prevented development of CR only in vitreous-inoculated rabbits. ON transection on the side opposite virus inoculation prevented CR regardless of the site of inoculation. These findings suggest that HSV-1 can leave the inoculated eye by multiple routes depending on the site of virus inoculation, but that virus reaches the retina of the contralateral eye via the optic nerve. PMID- 3030655 TI - Defining herpes simplex genes involved in neurovirulence and neuroinvasiveness. AB - In this discussion, strategies for physically mapping HSV gene functions associated with significant biologic effects are presented and discussed. They represent an application of molecular biological technologies to complex biologic systems, and are a beginning step in understanding the fundamental basis of herpetic disease. Future studies will involve precise identification of relevant genes, and studies of the important properties of products encoded by these genes. PMID- 3030656 TI - Herpes simplex virus isolation in chronic stromal keratitis: human and laboratory studies. AB - The corneal discs of 41 patients with scarring reminiscent of herpetic infection were organ cultured for HSV isolation. Of the 41 patients, 34 had a definite history of herpetic keratitis, from 10 of whom (29.4%) HSV was isolated. There were no clinical features which distinguished between these groups; there was however an indication that those from whom HSV was not isolated had been previously treated with substantial amounts of topical acycloguanosine. In three patients of 12 patients when the disc was separated into 7 parts using a punch technique, virus was isolated exclusively from those portions demonstrating clinical scarring. Electron microscopy (EM) demonstrated HSV particles in stromal cells in the cultured corneas of seven patients. In two of the patients no virus was detected prior to culture with EM. In one patient HSV antigen was not found using peroxidase-antiperoxidase (PAP) staining prior to subsequently positive organ culture. Studies were made to determine how HSV accedes to the corneal stroma using a murine model in which keratitis occurs by zosteriform spread of HSV following inoculation of the snout. Preliminary evidence using PAP staining indicates that the virus reaches the stroma at the same time as the epithelium, via the sensory nerves. Evidence of HSV persistence in anterior segments was obtained in the same model, in contrast to which no virus could be isolated following direct inoculation into the cornea. It is speculated that for virus to set up a longterm association with the stromal keratocyte, it must be introduced via the sensory nerve.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3030657 TI - HSV-induced reactivation: contribution of epinephrine after corneal iontophoresis. AB - A modified, bilateral iontophoresis technique was used to evaluate herpes simplex virus (HSV) reactivation and epinephrine distribution and concentration in full thickness corneas and in trigeminal ganglia of rabbits. Infectious virus was recovered from 58% of epinephrine-induced eyes 24-96 hours after iontophoresis. A histochemical adrenochrome oxidation method localized epinephrine exclusively in the corneal epithelium and anterior stromal lamellae after iontophoresis. Epinephrine introduced iontophoretically into the corneal epithelium and stroma did not reach the ganglion level as detected by the assay. Radiometric-enzymatic assay of corneal and ganglionic tissues demonstrated a transient increase in total corneal catecholamine levels from an average of 417 micrograms to an average of 778 micrograms ; concentrations returned to approximate control values 24 hours after iontophoresis. Ganglion total catecholamine levels increased from an average of 606 micrograms to an average of 1211 micrograms for a 12-hour period and remained elevated at 24 hours after iontophoresis. Epinephrine acting directly on the corneal epithelium and epinephrine metabolites at the ganglion level could act synergistically to induce ocular and ganglionic HSV reactivation. PMID- 3030658 TI - Study of HSV-1 DNA species from trigeminal ganglia of rabbits during acute and latent infections. AB - Recurrent herpetic keratitis remains a major cause of corneal blindness in developed countries. A fundamental unanswered question regarding herpes simplex virus infection concerns the relationship between the virus and host cell DNA during latency. In the present study DNA was extracted from trigeminal ganglia during both acute and latent infection following ocular inoculation. Extracted, purified DNA was utilized for transfection and for hybridization studies using a 32P-labeled HSV-1 DNA probe. DNA extracted during acute infection was complete, linear and non-integrated. Autoradiographic patterns of DNA isolated during latent infection were suggestive of two separate DNA species. PMID- 3030659 TI - Spread of HSV-1 to the mouse eye after inoculation in the skin of the snout requires an intact nerve supply to the inoculation site. AB - Infection of the eye following inoculation of herpes simplex virus on the skin of the snout was monitored using slit lamp examination of the eye, isolation of virus from eyewashings and identification of virus antigens in whole corneal epithelial sheets by peroxidase-antiperoxidase staining. Infection of the eye was prevented by removing a section of the sensory nerves which supply the inoculation site. This provided evidence that spread from the skin of the snout to the eye occurred via the nerves. PMID- 3030660 TI - Strain specificity of spontaneous and adrenergically induced HSV-1 ocular reactivation in latently infected rabbits. AB - Spontaneous ocular shedding and adrenergic induction of ocular shedding were examined in rabbits infected with ten strains of herpes simplex virus type 1 (HSV 1): McKrae, KOS, F, Rodanus, 17 Syn+, RE, E-43, SC-16, MacIntyre, and CGA-3. All ocular inoculations were with 50 microliter of HSV-1 with titers between 1-10 X 10(6) PFU/ml. All corneas, except those that received the McKrae strain, were scarified. Acute ocular infection was determined by slit-lamp biomicroscopy. Dendritic keratitis or geographic ulcers developed in all eyes of all rabbits within 10 days after ocular inoculation. All eyes of all surviving rabbits were swabbed for 20 consecutive days during days 20-39 postinoculation (PI). On PI day 19, no active lesions were present as judged by slit-lamp biomicroscopy. Ocular tear film was collected on a Dacron-tipped swab and placed on primary rabbit kidney cell monolayers. The cell monolayers were monitored for cytopathic effects consistent with HSV-1 infection. Spontaneous HSV-1 shedding was detected in some eyes from all groups of latently infected rabbits, except those infected with CGA 3. Spontaneous shedding (positive swabs/total swabs) of the other nine strains ranged from 0.7% to 15.7%. After PI day 42, the rabbit eyes received 6 hydroxydopamine by iontophoresis, followed for 5 days by topical application of 2% epinephrine. This procedure results in induced HSV-1 ocular shedding for a duration of 3-5 days in rabbits infected with the McKrae strain. In rabbits latently infected with KOS, F, RE, MacIntyre, and CGA-3, no induced HSV-1 shedding was detected.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3030661 TI - Non-small cell lung cancer. AB - Lung cancer stands as the most important malignant neoplasm in the United States because of its high prevalence, increasing incidence, high rate of mortality, and great potential for prevention through the control of cigarette smoking. The World Health Organization (WHO) classification of lung cancer identifies four major types: squamous cell carcinoma, adenocarcinoma, large cell carcinoma, and small cell carcinoma. These tumors are commonly divided into two groups based on differences in their biology and treatment: small cell (SCLC) and non-small cell carcinomas (NSCLC). This review analyzes NSCLC with a strong emphasis on the practical aspects of treatment. We give recommendations about smoking cessation and early diagnosis through screening of high-risk individuals. We review contemporary diagnostic and staging techniques in the context of the new international TNM system of staging. Subsequent discussions of treatment are based on this new staging system. We stress the pivotal role of surgery for the management of local disease, and in addition present the potential contributions of newer radiation therapy techniques. We examine chemotherapy in detail, including a review of the comparative activity of the available cytotoxic agents against NSCLC, the relative contribution of combination chemotherapy, and the role of surgical adjuvant treatment with either chemotherapy or immunotherapy. We advise that patients with NSCLC be treated under the aegis of modern clinical trials of new therapy whenever possible. When this is not possible, we recommend an individualized approach based on such factors as the patient's age, general state of health, cardiopulmonary status, psychosocial status, and personal system of values. PMID- 3030663 TI - The arenaviruses--an introduction. PMID- 3030662 TI - Neutrophil dysfunction and decreased leukotriene production in burned, septic rats. PMID- 3030664 TI - Pathology and pathogenesis of arenavirus infections. PMID- 3030665 TI - Bowenoid papulosis and squamous cell carcinoma of the genitalia: suspected sexual transmission. AB - The human papillomavirus causes a variety of genital lesions: condyloma acuminatum, bowenoid papulosis (carcinoma in situ), and squamous cell carcinoma. Only condylomata have been documented to be sexually transmitted. We report clinical and histopathologic evidence of suspected female to male transmission of bowenoid papulosis to the penis from a woman with condylomata acuminata, squamous cell carcinoma in situ, and focally invasive squamous cell carcinoma of the vulva. These findings indicate a need for thorough clinical and histopathologic evaluation of any anogenital lesions resembling condylomata and occurring in sexual partners. Conservative yet thorough destruction of bowenoid papulosis and squamous cell carcinoma in situ appears to be the treatment of choice. PMID- 3030666 TI - Familial polyposis coli. PMID- 3030667 TI - Cytomegalovirus pneumonitis and lobar consolidation. AB - This report describes the occurrence of localized lobar consolidation caused solely by cytomegalovirus infection in two heart transplant recipients. This highly atypical and previously unreported radiographic manifestation of cytomegalovirus pneumonitis underscores the need for vigorous diagnostic evaluation of immunosuppressed patients since localized pneumonitis in the immunocompromised host does not exclude the possibility of opportunistic infection. PMID- 3030668 TI - Therapeutic efficacy of itraconazole in systemic candidosis in guinea pigs. AB - Fifty non-immunocompromised guinea pigs were infected by the intravenous route with 8,000 blastospores of Candida albicans per gram body weight: 26 were treated orally with the excipient, 12 with itraconazole at 1.25 mg X kg-1 and 12 at 5 mg X kg-1, once daily for 14 days starting on the day of infection. Hematology was checked for all animals before infection and on days 7, 14 and 17 after infection. Histopathological examinations were done for 2 animals of each group on days 7, 14 and 17. The infection and the therapeutic efficacy were checked by clinical observation, at autopsy and by cultures of organs. Itraconazole was highly active at both concentrations, resulting in clinical cure, negativation of cultures, normalisation of the blood picture and absence of fungal elements and presence of only small remnants of lesions on days 14 and 17 in some organs. No drug-related side effects were observed. PMID- 3030669 TI - Effects of lincomycin on the immune system. AB - The effects of lincomycin on the immune system were studied on patients suffering from chronic bronchitis by means of phagocytosis, chemotaxis and natural-killer tests. The tests were performed before and 2, 4, 8, 12 and 24 h after administration of 600 mg lincomycin i.m. in a single dose. The results indicate that lincomycin stimulates phagocytosis, expressed as enhanced superoxide production (O2-), chemotaxis and the activity of natural killer cells, measured on the basis of their ability to perform a lysis on the K562 tumoral target labelled with 51Cr. The stimulating effects on chemotaxis appear already 2 h after the administration of the drug, while the same effect on phagocytosis and natural-killer activity occurs after 4 h. The maximal stimulating activity on all three parameters can be shown at 8 h and disappears at 24 h. PMID- 3030670 TI - [A sensitive reversed passive hemoagglutination assay for the estimation of urine human chorionic gonadotropin and its clinical applications]. PMID- 3030671 TI - Sequence diversity of gap junction proteins. AB - This paper summarizes our understanding of the molecular organization of gap junction proteins. There appear to be overall similarities in the organization of heart and liver junctions in terms of general domains, even though the molecular sizes of the two proteins are quite different. Sequence data on the amino terminal regions of these two proteins show 43% of the residues to be identical and 25% more to be homologous. The major intrinsic protein of lens (MIP), believed by many to be the lens-fibre junction protein, does not show such sequence homology with the known portions of junction proteins from either heart or liver. Yet the sequence of MIP, which is completely known, suggests a conformation for this molecule quite compatible with a junctional role. It thus appears that molecules potentially involved in junction formation will prove to form a rather diverse family, with special characteristics of organ-specific molecules that may well be related to their function. PMID- 3030672 TI - Topology of gap junction protein and channel function. AB - This paper presents recent results from this laboratory concerning the topological structure and function of the major 27 kDa gap junction protein. Immunological, biochemical and biophysical observations now provide evidence for the localization of the 27 kDa protein to gap junction structures, both in vivo and in vitro, and for the participation of the 27 kDa protein in channel conductance. PMID- 3030673 TI - The use of antibodies to gap junction protein to explore the role of gap junctional communication during development. AB - Antibodies raised against the major 27 kDa protein electrophoretically eluted from isolated gap junctions and affinity purified against the antigen have been used to explore the role of communication through gap junctions in the early amphibian and mouse embryos. In both species, injection of the antibodies into one cell completely blocks both dye transfer and electrical coupling between cells connected by gap junctions. In the amphibian embryo the generation of a communication-incompetent clone of cells leads to patterning defects in the region derived from the antibody-injected cell. In the mouse embryo, blocking cell-to-cell communication leads to decompaction of the communication-incompetent cells. The possible significance of these findings in relation to development in general and to the organization of the first transporting epithelia to appear during development is discussed. PMID- 3030674 TI - Molecular structure of the gap junctional channel. AB - The proteins in various gap junctional preparations from rodent liver have been analysed by two-dimensional peptide mapping and immunoblotting. Only the protein of relative molecular mass (Mr) 16,000 (16K) is found in all gap junctional isolates, and it is unrelated to the 27K protein. The absence of the 27K protein and any of its fragments from trypsin-treated preparations suggests that this protein does not directly contribute to gap junctional structure. Peptide mapping and immunoblotting of the 16K proteins isolated from various tissues and species and of the arthropod 18K protein present in gap junctional preparations from Nephrops norvegicus show that these proteins constitute a family of related junctional proteins. A site-specific antiserum raised against the N-terminal octapeptide of the 16K protein from mouse liver cross-reacts with all 16K and 18K forms of the junctional protein so far tested, suggesting that this particular antigenic determinant is highly conserved. Immuno-localization studies show that the N-terminus is most likely located on the cytoplasmic aspect of the junction and is available to Pronase digestion. PMID- 3030675 TI - [Multiple primary carcinoma of the large bowel]. PMID- 3030676 TI - Perianal Paget's disease. AB - Over the past 11 years (1974 to 1985) ten patients with perianal Paget's disease were treated. The average age was 64 years and half were male. Two patients were diagnosed as an incidental finding after hemorrhoidectomy and the remainder presented with a symptomatic perianal rash (itching and moisture) that averaged two years in duration. Physical examination in these patients demonstrated characteristic lesions (seven with erythematous or ulcerated, whitish gray lesions and one with a papillary lesion). Three patients presented with invasive carcinomas and, despite aggressive therapy, all developed metastatic disease. Two patients had local excisions with minimal margins and developed associated invasive cancers at four and ten years after diagnosis. The remaining five patients were treated by wide local excision and skin grafts. At present all are free of disease. The characteristic appearance of this lesion and its failure to respond quickly to conventional therapy should lead the clinician to obtain a biopsy which readily establishes the diagnosis. Experience confirms that wide local excision is adequate therapy, but adequate initial evaluation and close follow-up are necessary to identify other malignancies that may develop. PMID- 3030677 TI - Physiology of ileoanal anastomosis with ileal reservoir for ulcerative colitis and adenomatosis coli. AB - A physiologic and metabolic assessment was carried out on eight patients six months after total proctocolectomy with ileal reservoir for ulcerative colitis and familial adenomatosis coli. All patients were continent and able to defecate spontaneously, stool frequency ranging from two to five per 24 hours. Anal sphincter resting pressures (35 +/- 14 mmHg) and squeeze pressures (88 +/- 24.2 mmHg) were similar to those of a healthy population, with the exception of one patient's complaint of nocturnal mucous leakage per anus. Biopsies of the ileal mucosa of the reservoirs showed a mild inflammation in seven patients; in one a subtotal villous atrophy (plus glandular pattern) was found. Anthropometric measurements, lymphocyte counts, hemoglobin, albumin, transferrin, iron, B12, and folate were normal in all. In the majority of patients there was no evidence of bacterial overgrowth. Vitamin B12 absorption was reduced slightly in only one patient. Lipid absorption (as judged by the 14C-Triolein breath test) was abnormal in three patients. Fecal clearance of alpha 1 antitrypsin as protein losses index was abnormal in three patients. Bile acid malabsorption was the most important ileal dysfunction observed in the patients. PMID- 3030678 TI - Treatment of distal ulcerative colitis with beclomethasone enemas: high therapeutic efficacy without endocrine side effects. A prospective, randomized, double-blind trial. AB - Sixteen patients with 18 attacks of distal ulcerative colitis were treated randomly with either 0.5 mg topically administered beclomethasone dipropionate (BDP) or 5 mg betamethasone phosphate (BMT). The effect of the steroid enemas on adrenocortical function was examined by ACTH tests, which were performed before and 20 days after treatment. At completion of the trial, a marked suppression of the adrenocortical function was found in seven of eight patients treated for nine attacks with BMT but not in any patients in the BDP group (P less than 0.01). The mean posttreatment basal and stimulated plasma cortisol levels in the BMT group were significantly lower as compared with the BDP group. The overall therapeutic response assessed by score systems was comparable in the two treatment groups. It is concluded that, in the topical treatment of ulcerative colitis, BDP is preferable to BMT because it exerts an equal anti-inflammatory action without affecting adrenocortical function. PMID- 3030679 TI - [13C]methacetin breath test for evaluation of liver damage. AB - Methacetin undergoes rapid O-dealkylation by hepatic microsomal enzyme systems, and the resultant CO2 is present in the expired air. The rate of O-dealkylation of methacetin was assessed by the [13C]methacetin breath test in seven healthy volunteers and 30 patients with histologically proven chronic liver diseases. The 30-min recovery of orally administered [13C]methacetin as 13CO2 in the exhaled air was significantly reduced in patients with chronic aggressive hepatitis and in those with liver cirrhosis but not in patients with chronic persistent hepatitis or healthy controls. Patients with either advanced cirrhosis or hepatocellular carcinoma showed significantly lower values than those with well compensated cirrhosis. The levels in two patients with late primary biliary cirrhosis were reduced. These results show that the severity of liver damage can be effectively evaluated by [13C]methacetin breath test. In addition, this test is simple, safe, and time efficient. PMID- 3030680 TI - The human growth hormone gene locus: structure, evolution, and allelic variations. AB - Genomic clones containing the closely related genes for human growth hormone (hGH) and chorionic somatomammotropin (hCS) were obtained from genomic bacteriophage lambda and cosmid libraries. The entire GH/CS chromosomal locus was reconstructed utilizing overlapping restriction fragments characterized from the isolated clones. The hGH/hCS locus contains two GH genes and three CS genes spanning 48 kb of DNA in the order: 5'-(hGH-1/hCS-5/hCS-1/hGH-2/hCS-2)-3', confirming analysis of cosmid clones obtained from a different human library (Barsh et al., 1983). To complete the characterization of the hCS genes, the nucleotide sequence of the hCS-5 gene was determined. Sequence analysis revealed a mutation of the 5' splice site at the exon II-intron B boundary, suggesting that the hCS-5 gene is a pseudogene. The nucleotide sequence of an allelic variant of the hCS-2 gene was determined and found to contain a single amino acid substitution and the deletion of a single codon. The hGH/hCS gene locus was further characterized by the localization of at least 27 Alu-type repetitive sequences and identification of three unique sequences in the vicinity of several hGH and hCS genes which define the probable breakpoints of the evolutionary duplication units. These data, combined with the nucleotide sequences of all five GH and CS genes, indicate that the hGH/hCS gene locus has evolved by duplication mechanisms. Evidence for the occurrence of at least one gene conversion event involving the hCS-1 gene precursor and the hCS-2 gene was found, indicating that the hGH/hCS gene locus has evolved by concerted mechanisms. The structure of the hCS genes is discussed in light of recent studies of CS genes from other mammalian species. PMID- 3030682 TI - [Hormone-sensitive stage in the development of retinal pigment epithelium in genotypically normal rats]. PMID- 3030681 TI - [Effect of thymosin on afferent transmission in the somatosensory system of Macaca rhesus]. PMID- 3030684 TI - [Gyrase inhibitor: central nervous system side effects]. PMID- 3030683 TI - [Photoexcitation of electron orbits in the magnesium atom and (Na+-K+) ATPase functioning]. PMID- 3030685 TI - [HIV antibodies following CMV immunoglobulin prophylaxis]. PMID- 3030686 TI - Dietary fibre and the gut microflora--their effects on toxicity. PMID- 3030687 TI - Biochemical mechanisms of cephaloridine nephrotoxicity. PMID- 3030688 TI - Changes in abilities of producing and scavenging oxygen radicals in cephaloridine induced nephrotoxicity in rats. PMID- 3030689 TI - Extrapulmonary oat-cell carcinoma of the tonsil. PMID- 3030690 TI - Neurophysiological investigations in multiple nerve crush injury. PMID- 3030691 TI - Relationship between thresholds detection of vibratory sensation and sensory nerve electrophysiology. PMID- 3030692 TI - Bifunctional role of transforming growth factor-beta during granulosa cell development. AB - Regulatory actions of transforming growth factor-beta (TGF beta) on granulosa cell function were analyzed in cells cultured from the ovaries of diethylstilbestrol-implanted rats. In the presence of a suboptimal concentration of FSH (5 ng/ml) that increased LH receptors by 100-fold during a 72-h culture, TGF beta augmented this response in a dose-dependent manner with a maximal effect at 16 pM. In contrast, the growth factor inhibited the LH receptor response to an optimal dose of FSH (50 ng) by up to 50% and was inactive in the absence of gonadotropin. TGF beta also enhanced the formation of cAMP by 5 ng FSH and partially inhibited the effects of higher FSH concentrations. However, the actions of TGF beta were more prominent on LH receptor induction than on cAMP production with either low or high amounts of FSH. In addition, TGF beta had little effect on cAMP production stimulated by cholera toxin or forskolin, but amplified the actions of these ligands as well as that of 8-bromo-cAMP on LH receptor expression. TGF beta also modulated the steroidogenic activity of the granulosa cells, with increased production of progesterone in response to 5-100 ng FSH. The bifunctional actions of TGF beta on FSH-induced LH receptor formation were observed throughout a 96-h culture period. However, the presence of the growth factor was not required for the first 24 h of culture, indicating that TGF beta alters the later events involved in LH receptor formation. TGF beta augmented the stimulatory actions of 5 ng FSH on LH receptors in the absence or presence of insulin, but its inhibitory effect on these receptors was only observed in cells treated with insulin. These results indicate that TGF beta modifies FSH action during granulosa cell development in a biphasic manner. TGF beta can exert stimulatory or inhibitory effects depending upon the concentration of FSH and the presence of insulin, and these are due to alterations in cAMP action as well as cAMP production. Autocrine and/or endocrine actions of TGF beta during granulosa cell differentiation may be involved in the processes of follicle selection and development. PMID- 3030693 TI - Thyroid effects on adenosine 3',5'-monophosphate levels and adenylate cyclase in cultured neuroblastoma cells. AB - Using neuroblastoma cells as a model of developing neurons, we have tested the hypothesis that thyroid hormones alter cAMP metabolism. Neuroblastoma cells were grown in serum-free defined medium for 48 h with or without thyroid hormones. Treatment with 20-200 nM 3,5,3'-triiodo-L-thyronine (T3) increased the accumulation of cAMP by intact cells without altering growth, gross morphology, or DNA or protein content. The increase in cAMP accumulation could be detected 5 h after the addition of T3 and was abolished by the addition of cycloheximide. The maximum stimulation produced by prostaglandin E1 was increased in T3 cells without a significant alteration of the half-maximal concentration. T4 and D-T3 in concentrations up to 20 microM did not increase cAMP accumulation. Adenylate cyclase activity in response to forskolin, guanine nucleotides, and stimulatory hormones was increased in purified membranes from cells grown in T3, suggesting that increased adenylate cyclase is probably the major mechanism of the observed response to thyroid hormone. PMID- 3030694 TI - Differential induction of hepatic estrogen receptor and vitellogenin gene transcription in Xenopus laevis. AB - Low levels of estrogen receptor (200-500 per cell) are present in the liver of hormonally naive male Xenopus. However, administration of estradiol results in a rapid 2- to 5-fold increase in cellular estrogen receptor content concurrent with the de novo transcriptional activation of the genes for the yolk protein precursor vitellogenin. Studies on Xenopus embryogenesis suggest that estrogen receptor induction is required for the activation of vitellogenin transcription. The purpose of the present study was to examine the mechanism of estrogen receptor induction in male Xenopus liver. The experimental protocol used 4 hydroxytamoxifen, an antiestrogen with a high affinity for the estrogen receptor, to inhibit the effects of estradiol. Changes in estrogen receptor content were then determined through the use of an exchange assay. 4-Hydroxytamoxifen alone suppressed the level of estrogen receptor from 800 sites per cell in hormonally naive animals to 250 sites per cell. Administration of estradiol 24 h after the antiestrogen resulted in the induction of estrogen receptor to a level equivalent to that found in control animals (800 sites per cell). However, under the same conditions, estradiol was unable to overcome the antiestrogen inhibition of vitellogenin gene transcription. Although 4-hydroxytamoxifen displayed a high affinity for the hepatic estrogen receptor, it did not inhibit the binding of estradiol to a middle affinity cytoplasmic estrogen-binding protein. These results suggest that different mechanisms are involved in the induction of estrogen receptor and vitellogenin gene transcription. PMID- 3030695 TI - Decreased activity of adrenal S-adenosylmethionine decarboxylase in rats subjected to dopamine agonists, metabolic stress, or bodily immobilization. AB - The activity of S-adenosylmethionine decarboxylase (SAM-DC) has been measured in the adrenal gland of rats given treatments that are known to result in increased activity of ornithine decarboxylase in this organ. In contrast to the effects of the dopamine agonists piribedil and apomorphine on the latter enzyme, the administration of these drugs caused decreases of SAM-DC in both parts of the gland. After piribedil the activity decreased rapidly to a minimum at 2-4 h, with recovery by 6 h. The stress of immobilization or the administration of insulin or 2-deoxyglucose (2-DG) also decreased adrenal SAM-DC activity. The results contrast with those observed in other rat tissues where SAM-DC is generally induced by treatments that induce ornithine decarboxylase. Denervation of the adrenal gland did not clearly affect the reduction in adrenomedullary SAM-DC after 2-DG. Hypophysectomy resulted in reduced SAM-DC activity in both adrenal medulla and cortex; the activity could be restored by giving the animals 2 IU ACTH daily for 4 days. These changes in activity were parallelled by changes in immunoreactive protein. 2-DG did not decrease SAM-DC in hypophysectomized rats receiving maintenance ACTH dosage. This indicates the presence of hormonal control over the activity of SAM-DC in the adrenal medulla and cortex. Acute administration of an additional 10 IU ACTH to hypophysectomized rats on maintenance dosage of ACTH resulted in decreased SAM-DC activity in both adrenal medulla and cortex. These decreases were not abolished by inhibition of corticosteroid synthesis with metopirone. PRL and GH had no significant effect on adrenal SAM-DC activity of hypophysectomized rats. The reduction of SAM-DC activity in both parts of the gland of hypophysectomized rats with administration of (Bu)2cAMP suggests that cAMP may mediate the decreases in SAM-DC caused by the above treatments. PMID- 3030696 TI - A putative mediator of insulin action which inhibits adenylate cyclase and adenosine 3',5'-monophosphate-dependent protein kinase: partial purification from rat liver: site and kinetic mechanism of action. AB - A novel putative mediator of insulin action which acts to inhibit adenylate cyclase and cAMP-dependent protein kinase has been purified from livers of insulin-treated streptozotocin-diabetic rats. It was increased by short term (5 min) insulin injections in vivo and purified several thousand-fold by Sephadex and HPLC. Its mol wt was somewhat larger (2500) than previous mediators identified, and it was more hydrophobic in character. Its mechanism of action or adenylate cyclase was determined and found to be chiefly directed against the catalytic subunit. Its action on the cAMP-dependent protein kinase was found to be competitive with regard to protein substrate, but noncompetitive with regard to ATP and cAMP. Its relationship to other putative insulin mediators and the mechanism of insulin action is discussed. PMID- 3030697 TI - Hypophysial-portal plasma levels, median eminence content, and immunohistochemical staining of corticotropin-releasing factor, arginine vasopressin, and oxytocin after pharmacological adrenalectomy. AB - Changes in immunostaining, median eminence content, and secretion into the hypophysial-portal circulation of immunoreactive CRF (irCRF), arginine vasopressin (irAVP), and oxytocin (irOT) were directly evaluated after pharmacological adrenalectomy (PHADX). Mean circulating levels of ACTH rose from 270 +/- 57 (+/- SE) to 1560 +/- 283 pg/ml after 72 h of treatment with metyrapone and aminoglutethimide. Initially, hypophysial-portal plasma irCRF levels decreased to 52.6% (12 h) and 21.7% (24 h) of control levels (230 +/- 41 pg/ml). Accompanying changes in the patterns of CRF immunostaining in the paraventricular nuclei (PVN) or in median eminence irCRF content at 24 h did not parallel alterations in portal plasma irCRF levels at this time. By 72 h posttreatment, portal irCRF levels were elevated 2.2-fold, while the number of detectable CRF positive perikarya in the PVN increased 3.0-fold. The mean hypophysial-portal plasma irAVP concentration was unchanged from the control value (1312 +/- 287 pg/ml) at 12 h, but was only 34.9% of the control value at 24 h. Inverse changes in median eminence irAVP content were noted at these times, whereas the number of AVP-immunostained cells exhibited a tendency toward an increase at 24 h, in parallel with significantly increased content. By 72 h post-PHADX, portal irAVP, median eminence irAVP content, irAVP immunostaining intensity, and AVP immunopositive cell number were elevated. Approximately 64% of CRF-positive perikarya in the parvocellular PVN costained for AVP at this time, whereas no colocalization was evident in untreated rats. These changes were prevented by corticosterone replacement. irOT staining intensity, irOT-positive cell number, median eminence irOT content, and portal plasma irOT concentration remained stable at all times examined. We conclude that: removal of adrenal steroids by PHADX results in a sequence of changes in CRF and AVP within the PVN (as determined by immunocytochemistry) and the median eminence (as determined by peptide content) similar to those observed after surgical adrenalectomy; after steroid removal the secretion of both irCRF and irAVP changes in a biphasic manner characterized by reduced secretion at 24 h and greatly enhanced secretion at 72 h; neither immunostaining nor median eminence content alone proved to be a reliable index or secretory activity during the initial phases of steroid blockade; and the hypophysiotropic OT system of normal male rats appears to be insensitive to adrenal steroid influences. PMID- 3030699 TI - Immunoreactive corticotropin-releasing factor in rat plasma. AB - Immunoreactive ACTH (I-ACTH) levels in the rat anterior pituitary and plasma, and immunoreactive corticotropin-releasing factor (I-CRF) concentrations in the median eminence (ME) and plasma were determined after adrenalectomy and in insulin-induced hypoglycemia. I-CRF was detected in plasma from normal rats (mean +/- SD, 5.6 +/- 0.9 pg/ml; n = 6). Gel filtration chromatography of I-CRF from pooled plasma of these rats revealed a single peak which eluted in the position of authentic rat CRF. I-CRF levels in ME and I-ACTH levels in anterior pituitary decreased immediately after adrenalectomy, then gradually increased to high levels 14 days after surgery. Plasma I-CRF and I-ACTH concentrations increased immediately after surgery, slightly decreased to near the control levels at 24 h, and then increased to high concentrations 14 days after surgery. Plasma and ME I CRF levels 14 days after adrenalectomy, followed by daily dexamethasone replacement, were almost the same as control levels. In insulin-induced hypoglycemia, plasma I-ACTH and I-CRF concentrations increased and ME I-CRF content decreased at 30 and 60 min. These results suggest that plasma I-CRF levels reflect changes in hypothalamic CRF levels. PMID- 3030698 TI - Liver angiotensinogen synthesis and release during captopril treatment in sodium depleted rats. AB - In vivo generation of angiotensins depends upon both plasma renin and angiotensinogen concentrations. Those factors which may influence hepatic angiotensinogen synthesis and release were examined. We have evaluated in vivo the effects of converting enzyme inhibition on several plasma renin-angiotensin system components, and, using an in vitro preparation of liver slices, we also investigated the effects of converting enzyme inhibition on the synthesis and release of hepatic angiotensinogen. Angiotensinogen concentrations were determined by two different methods. The first was an indirect enzymatic assay which measures the amount of angiotensin I liberated from plasma by an excess of renin. The second was a direct RIA that measures both angiotensinogen and its inactive residue the des-angiotensin I-angiotensinogen. The difference between the methods represents the circulating levels of des-angiotensin I angiotensinogen. Captopril administration in sodium-depleted rats increased plasma concentrations of renin, des-angiotensin I-angiotensinogen, and angiotensin I and decreased plasma angiotensinogen concentration measured by both methods. Plasma des-angiotensin I-angiotensinogen was significantly correlated to plasma renin concentration, which suggests an increase in the consumption of angiotensinogen when the renin secretion is extremely increased. The angiotensinogen liver content and in vitro angiotensinogen release were decreased in sodium-depleted rats treated with a converting enzyme inhibitor, and these parameters were negatively correlated to in vivo plasma levels of renin, angiotensin I, and des-angiotensin I-angiotensinogen. They were positively correlated to plasma angiotensinogen concentration measured by the indirect assay. These data suggest that captopril administration during sodium depletion has two simultaneous effects: it increases angiotensinogen consumption and second, decreases angiotensinogen production and release. PMID- 3030700 TI - Regulatory effect of lithium on thyroxine metabolism in murine neural and anterior pituitary tissue. AB - The conversion of T4 to T3 in the brain and anterior pituitary gland contributes significantly to the T3 content of these tissues and appears to be an important modulator of thyroid hormone action. In the present study, the antimanic agent lithium was demonstrated in cultured neural and pituitary tissue to have a significant inhibitory effect on the activity of low Km (type II) iodothyronine 5'-deiodinase (I5'D), the enzyme mediating T3 formation. At medium lithium concentrations of 3.3-5 mM, 15'D activity was decreased 44 +/- 3% (P less than 0.001) in the NB41A3 mouse neuroblastoma cell line and 48 +/- 2% (P less than 0.001) in the GH3 rat pituitary tumor cell line. This inhibitory effect was only observed in intact cells. Significant inhibition of this enzymatic process was also noted in the anterior pituitary gland of thyroidectomized rats injected 3-24 h earlier with either 4 or 10 mmol/kg BW LiCl. This decrease in low Km I5'D activity was accompanied by significant decreases in the serum T3 concentration and the pituitary nuclear T3 content. Renal high Km (type I) I5'D activity was unaffected by lithium administration. These studies demonstrate that lithium, an agent of proven therapeutic benefit in patients with manic-depressive illness, can affect changes in T4 metabolism and cellular T3 content in neural and anterior pituitary tissue. Given the prominent mood changes that occur in patients with disordered thyroid function, this finding suggests that the therapeutic benefits of lithium in affective illness may be derived in part from alterations in thyroid hormone economy in the brain. PMID- 3030701 TI - Rat uterine progesterone receptor analyzed by [3H]R5020 photoaffinity labeling: evidence that the A and B subunits are not equimolar. AB - The hormone-binding components of the rat uterine progesterone receptor were investigated by the methods of [3H]R5020 photoaffinity labeling and sodium dodecyl sulfatepolyacrylamide gel electrophoresis analysis. Two specifically labeled peaks were observed at mol wt of 85,600 +/- 1,200 and 109,600 +/- 1,200 (n = 31), resembling the A and B progesterone receptor components previously described in other systems. However, in contrast to the equimolar ratio reported in other systems, the level of subunit A observed was consistently greater than that of B (A/B ratio = 3.2 +/- 0.3; n = 31). The unusual A/B ratio prompted a complete validation of the photolabeling procedure in this system. Although the levels of specific binding increased, there was no change in the A/B ratio with varying [3H]R5020 concentrations (5-80 nM) or with time of UV exposure (0.5 min to 3 h). Although adsorption to hydroxylapatite indicated that specific [3H]R5020 binding was reduced by 72.0 +/- 6.4% within 5 min of UV exposure, relabeling the irradiated preparations with [3H]R5020 resulted in little change in specific [3H]R5020 binding. TLC analysis of [3H]R5020 (Rf = 0.48 +/- 0.01; n = 4) after irradiation demonstrated rapid photolysis resulting in a 94.3 +/- 2.5% (n = 3) loss of authentic [3H]R5020 within 5 min. After photolysis, at least two new tritiated products were recovered with Rf values of 0.20 +/- 0.03 and 0.72 +/- 0.02. Analysis by adsorption to hydroxylapatite indicated that the photolysis products competed for specific [3H]R5020-binding sites in cytosol with only 10 fold lower relative binding activity than authentic R5020. Thus, these compounds probably account for the increase in specific photolabeling of the A and B peaks achieved when UV exposure is prolonged from 5 to 30 min. Further study indicated that the A/B subunit ratio in this system was not changed under a variety of in vitro conditions, including the absence or presence of molybdate, sulfhydryl protective reagents (dithiothreitol and thioglycerol), protease inhibitors (phenylmethylsulfonylfluoride and leupeptin), glycerol (0%, 10%, and 30%, vol/vol), or 1.5 mM EGTA, or after precipitation with 40% ammonium sulfate. This consistency of the A/B ratio under a wide variety of adverse in vitro conditions suggests that in vitro artefacts may not account for the ratio's deviation from unity. Estrogen withdrawal (48 h) enhanced by progesterone treatment (0.5 mg for 24 h) resulted in only a modest reduction in the A/B ratio to 1.9 +/- 0.1.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3030702 TI - Aromatase activity in a rat Leydig cell tumor. AB - Male Wistar-Furth rats bearing the transplantable LTW(m) Leydig cell tumor have elevated serum estradiol (E2) concentrations. We measured the ability of these tumors to aromatize testosterone (T) to E2 by two methods. First, tumor minces were incubated with [7-3H]T, and the resultant [3H]E2 and [3H]estrone were purified and measured. In addition, tumor cell cultures were incubated with [1 beta-3H]T, and the resultant [3H]H2O was determined as a measure of aromatization. Tumor minces aromatized more actively than normal rat testicular tissue (3.30 +/- 0.15% of the T added was converted to E2 by the tumor vs. 0.30 +/- 0.25% by normal testis). Most of the aromatizing actitivity was localized to the microsomes. Using cell cultures the maximum velocity was 6.1 pmol/h X 5 X 10(5) cells, and the Km was 98 nM. In neither minces nor cell cultures were we able to show stimulation of aromatization with hCG, (Bu)2cAMP, or phorbol esters, although we could show stimulation by these agents in normal testicular cells. We were unable to inhibit the aromatase activity with human beta-endorphin or stimulate it with naloxone. However, we were able to inhibit the aromatase activity with 4-hydroxy-4-androstene-3,17-dione. We conclude that the LTW(m) rat Leydig cell tumor has an active autonomous aromatase system that is not responsive to compounds affecting the adenylate cyclase-cAMP system. It can be inhibited by 4-hydroxy-4-androstene-3,17-dione, a competitive-suicide inhibitor of the aromatase enzyme(s). PMID- 3030703 TI - 12-O-tetradecanoyl phorbol-13-acetate potentiates adenosine 3',5'-monophosphate mediated chorionic gonadotropin secretion by cultured human choriocarcinoma cells. AB - The activation of protein kinase C and adenylate cyclase in the regulation of human CG (hCG) synthesis by cultured BeWo choriocarcinoma cells was studied. Both cholera toxin (CT), which activates adenylate cyclase, and 12-O-tetradecanoyl phorbol-13-acetate (TPA), a protein kinase C activator, stimulated the secretion of hCG in a dose-dependent manner. During a 72-h culture, stimulation with the maximal effective concentration of CT (1.0 ng/ml) exerted a more pronounced increase (16-fold) in hCG synthesis than TPA (10 ng/ml) (2.8-fold), whereas the inactive phorbol ester 4 alpha-phorbol 12,13-didecanoate (100 ng/ml) was ineffective. When added together, TPA potentiated the effect of CT on hCG secretion (from 16- to 27-fold) and cAMP accumulation (from 36- to 54-fold) in the medium. TPA (1.0 ng/ml) also caused a 2.0-fold increase in basal cAMP production after 72 h. Time-course studies indicated that the effect of TPA on CT induced cAMP and hCG productions became significant at 45 min and 6 h, respectively, from the beginning of stimulation. Proliferation of cells was not responsible for the responses, since the treatments only slightly increased the total protein content and did not alter the rate of incorporation of C3H3 methylated thymidine of the cells. Our results demonstrate that TPA potentiates CT-induced cAMP and hCG production in cultured human choriocarcinoma cells. PMID- 3030704 TI - Differential regulation of brain and pituitary corticotropin-releasing factor receptors by corticosterone. AB - The regulatory actions of CRF during the neuroendocrine response to stress are mediated by specific receptors within the nervous system and the anterior pituitary gland. Glucocorticoids exert negative feedback inhibition on ACTH secretion by interacting at the pituitary corticotrophs and the central nervous system. To determine whether glucocorticoids influence ACTH secretion by regulating the concentration of CRF receptor sites, binding of [125I]Tyr-oCRF to pituitary and brain membrane-rich particles was studied after glucocorticoid treatment. Corticosterone administration (0.5-150 mg/day) for 1-4 days in adult male rats caused a dose-dependent decrease in the number of CRF receptors in the anterior pituitary in parallel with the reduction in ACTH secretion. In the brain, binding studies in membrane-rich fractions or by autoradiography in slide mounted frozen sections revealed no changes in CRF receptors in the cortex, hippocampus, amygdala, septal area, and olfactory bulb, although circulating corticosterone levels were higher than during stress. The selective down regulation of anterior pituitary CRF receptors after corticosterone administration, without alterations in brain CRF receptors, is similar to the change in CRF receptors previously reported after adrenalectomy and indicates that receptor regulatory mechanisms in secretory cells differ from those in neural tissue. Furthermore, the decrease in pituitary CRF receptors after physiological increases in circulating glucocorticoids may contribute to the inhibitory effects of adrenal steroids on ACTH secretion. PMID- 3030705 TI - Beta-adrenergic receptors, glucagon receptors, and their relationship to adenylate cyclase in rat liver during aging. AB - The beta-adrenergic and glucagon receptor-binding capacities in rat livers from 6 27 months of age were measured to investigate the mechanism of a previously observed rise in beta-adrenergic stimulated adenylate cyclase with increasing age. There was no concomitant increase in glucagon-stimulated adenylate cyclase. In the present study neither glucagon-binding capacity nor glucagon-stimulated adenylate cyclase changed with age. In contrast, the beta-adrenergic receptor capacity, measured in the same membranes by [125I]iodopindolol binding, increased nearly 3-fold from 6.6 +/- 0.6 fmol/mg at 6 months to 19.1 +/- 3.3 fmol/mg at 18 19 months. The increase was directly proportional to the maximum isoproterenol stimulated adenylate cyclase activity in livers of rats up to 19 months of age. By 24-27 months the binding capacity had increased to 24.9 +/- 3.3 fmol/mg, but there was no further increase in adenylate cyclase activity. Thus, there appeared to be a beta-receptor-adenylate cyclase uncoupling in livers from the senescent animals (25-27 months). The defect could not be demonstrated by studies examining isoproterenol competition of [125I]iodopindolol from agonist-induced high affinity sites on the membranes, a procedure that examines receptor-Ns protein coupling. Activation of adenylate cyclase by the nonhormonal stimulators F- and forskolin did not change with age, indicating that the catalytic unit was not a limiting factor. Since the relationship between the glucagon receptor and adenylate cyclase also remained unaltered, the uncoupling apparently lies in an alteration of the interaction between the beta-adrenergic receptor and the guanine nucleotide-sensitive Ns protein. PMID- 3030706 TI - Mutually independent effects of adrenocorticotropin on luteinizing hormone and testosterone secretion. AB - In this study, we examined the effect of ACTH on the sensitivity of the testes to gonadotropin and determined the role of the testosterone (T) negative feedback system in mediating the inhibitory effect of ACTH on LH secretion in adult male rats. ACTH infusion for 3 days reduced basal levels of serum T and the T response to GnRH, but did not alter basal levels of serum LH (immunoreactive) or the LH response to GnRH. These effects required the presence of the adrenal glands. Infusion of corticosterone (B) at a dose that increased serum B concentrations 9 fold had an effect similar to that of ACTH on basal serum T levels and the serum T response to GnRH. Basal levels of serum LH and the serum LH response to GnRH were not affected by B administration. These data suggest that ACTH administration reduces the sensitivity of the testes to LH, resulting in a lower basal level of T and a reduced T response to GnRH. This effect was independent of basal serum LH levels or the LH response to GnRH. It appears that B mediates the effect of ACTH on testicular sensitivity to gonadotropin. In another experiment, ACTH administration for 4 days did not alter serum LH values, but reduced serum T levels in sham-castrated male rats. In contrast, ACTH treatment blunted the increase in serum LH after castration by day 2 of treatment, despite the absence of detectable levels of serum T within 6 h after castration. These data suggest that T is not essential for the inhibitory effect of ACTH on LH secretion to occur. They do not support the hypothesis that ACTH enhances the sensitivity of the hypothalamus and/or pituitary to the negative feedback effects of T. PMID- 3030707 TI - Adenosine 3',5'-monophosphate primes the secretion of follicle-stimulating hormone. AB - We investigated whether cAMP acts as a mediator for LHRH in either its immediate FSH release action or its self-priming action. Pituitary pieces from cyclic female rats were superfused in vitro in the presence of Bu2cAMP, 8-bromo-cAMP, or forskolin or used as controls. For pituitary pieces from proestrous rats, the first significant increase in the baseline FSH secretion rate occurred after approximately 90 min of exposure to elevated cAMP resulting from forskolin treatment. By comparison, in the same system LHRH caused a 3-fold increase in FSH secretion during a 10-min exposure to the peptide. In contrast to its ineffectiveness as a secretagogue, cAMP elevation resulted in a several-fold augmentation of both LHRH- and elevated K+-stimulated FSH secretion from pituitary pieces from proestrous, but not estrous, rats; for these experiments, superfusion with a cAMP analog or forskolin for varying times preceded a 10-min pulse of either 8 nM LHRH or 47 mM K+. Augmentation of K+-stimulated secretion was evident after 30 min of cAMP elevation. Priming of LHRH-stimulated FSH secretion required 30-90 min of pretreatment with cAMP; longer exposures to cAMP analogs or forskolin were coincident with greater potentiation. Cycloheximide prevented Bu2cAMP augmentation of LHRH-stimulated FSH secretion. These data show that cAMP does not mimic the FSH release action of LHRH, but does augment LHRH- or K+-stimulated FSH secretion with characteristics that lead us to suggest that cAMP mediates, at least in part, the self-priming function of LHRH. PMID- 3030708 TI - The Syrian hamster pineal gland responds to isoproterenol in vivo at night. AB - Failure of isoproterenol (ISO) injections to raise pineal melatonin content has generated doubt about beta-adrenergic control of the melatonin rhythm in Syrian hamsters. However, the effect of ISO injected at night after light-induced reduction of pineal melatonin has not been reported. In this study, light exposure began at 6 1/4 h into one (normally 10-h) dark phase. The hamsters were injected with either ISO (1 mg/kg) or vehicle 15 min later when pineal melatonin content was low. Light exposure continued. Two h after ISO but not vehicle injection, pineal melatonin content rose more than six-fold. In other animals injected at the end of the usual light phase then kept in light for 2 h, pineal melatonin was equally low after ISO or vehicle injection. The Syrian hamster pineal gland can respond in vivo to a beta-adrenergic agonist injected at the physiologically relevant time of the normal nocturnal melatonin surge. This finding, taken together with the previously reported inhibition of the endogenous nocturnal melatonin surge with a beta-blocking drug, suggests that a beta adrenergic mechanism controls the hamster pineal melatonin rhythm. PMID- 3030709 TI - Effect of synthetic atrial natriuretic polypeptide on hemorrhage-induced adrenocorticotropin secretion of the rat. AB - The effect of synthetic alpha-human atrial natriuretic polypeptide (alpha-hANP) on the in vivo and in vitro release of ACTH and corticosterone was examined. In the in vivo study ACTH and corticosterone responses to rapid 2-ml/rat hemorrhage were measured in sixteen conscious rats after alpha-hANP administration. The hemorrhage increased plasma ACTH and corticosterone concentrations in the control group of rats (p greater than 0.01). ANP inhibited hemorrhage-induced ACTH secretion (p less than 0.05), but the plasma corticosterone response was not affected. In the in vitro study a high concentration of ANP (1 microM) reduced basal corticosterone secretion from the isolated rat adrenal gland (p less than 0.05), but the response to ACTH (10 ng/ml) and dibutyryl cyclic AMP (0.5 mM, 5.0 mM) was not affected. Our data suggest that ANP inhibits hemorrhage-induced ACTH secretion from the anterior pituitary but inhibits corticosterone secretion from the adrenal gland very weakly. PMID- 3030710 TI - Maintenance of pancreatic endocrine cells of the neonatal rat: VII. The effect of 3-amino-3-deoxyglucose on insulin secretion from perifused B cells. AB - Monolayer cultures of the pancreas of the neonatal rat were maintained in TCM 199 medium, supplemented with 5.5 mM glucose, with or without 5 mM 3-amino-3 deoxyglucose, and perifused to examine the changes which occurred in the insulin secretory response during culture. On day 0, B cells showed a monophasic insulin secretion in response to 16.7 mM glucose, whereas in the presence of 200 nM 12-o tetradecanoyl phorbol-13-acetate, 40 microM lysophosphatidylcholine, 10 microM forskolin or 1 mM 3-isobutyl-1-methylxanthine, the same dose of glucose stimulated insulin secretion in a biphasic fashion. Under culture conditions without 3-amino-3-deoxyglucose, the response to glucose totally disappeared after 7 days, and that to 10 mM of either leucine or 2-ketoisocaproate was as low as that of day 0. In contrast, B cells that had been cultured for 7 days in medium with 3-amino-3-deoxyglucose showed an adult-like biphasic pattern in response to glucose. When stimulated by glucose at a linear gradient concentration running from 0 to 20 mM, the B cells responded to increasing concentrations of glucose in a dose-dependent fashion. Further, the response of cAMP to glucose was increased by adding forskolin or 3-isobutyl-1-methylxanthine, which also enhanced the secretion of insulin under either a step-wise or slow-rise stimulation with glucose. The effect of 12-o-tetradecanoyl phorbol-13-acetate was also outstanding. Likewise, the addition of either leucine or 2-keptoisocaproate induced a striking increase in the secondary phase secretion as well as promoting the rates of glutamine oxidation within the cells. In conclusion, it is suggested that the high response to a wider variety of stimuli may represent the reaction of neonatal B cells to the cultural milieu rather than a process of physiological development, and these effects exhibited by 3-amino-3-deoxyglucose would be related to a change in the constituents of glycoproteins in the cells. PMID- 3030711 TI - Biological activities of sarcosine1-angiotensin I in man. AB - In order to examine whether substrate specificity of angiotensin-converting enzyme (ACE) exists or not for N-terminal substituted angiotensin I (ANG I) in man, biological activities of sarcosine1-angiotensin I (Sar1-ANG I) and the effects of an ACE inhibitor, captopril, on the Sar1-ANG I activities were studied in 5 normal men. The following 3 experiments were done at 1 week intervals. Sarcosine1-angiotensin II (Sar1-ANG II) was infused iv at a rate of 5 pmol/kg X min from 0900 h to 0930 h in 5 normal men in a recumbent position. Blood pressure rose remarkably and the average increment was 38/31 mmHg at 30 min (p less than 0.001). Average duration of the pressor action after the cessation of the infusion (T) was 40 min for systolic and 50 min for diastolic and much longer than T of isoleucine5-angiotensin II. Plasma renin activity (PRA) decreased (p less than 0.01) and plasma aldosterone (PA) increased significantly (p less than 0.01). Sar1-ANG I was infused iv at a rate of 5 pmol/kg X min from 0900 h to 0930 h. Blood pressure rose to the same extent as in (1) (p less than 0.001). T was 40 min for both systolic and diastolic and much longer than T of ANG I in man. PRA decreased (p less than 0.01) and PA increased (p less than 0.01) significantly. Oral 100 mg captopril was given at 08:00 h and Sar1-ANG I was infused iv at a rate of 5 pmol/kg X min from 09:00 h to 09:30 h.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3030712 TI - Acute endocrine effects of a single administration of methylmercury chloride (MMC) in rats. AB - The acute effects of methylmercury chloride (MMC) on the endocrine functions were investigated with doses too small to cause any typical neurological dysfunctions. The hormones included PRL, LH, TSH, ACTH, corticosterone (Bk), testosterone (TLI), total thyroxine (T4) and free thyroxine (free T4). The changes in serum hormone levels from 1 hour through 10 days after a single injection of MMC (12 mg/kg s.c.) (Exp. 1), and dose-response relationships between MMC doses (2 to 16 mg/kg s.c.) and the serum hormone levels at 25 hours after MMC injection (Exp. 2) were examined. The acute effects revealed, which were all reversible, are summarized as follows; MMC might directly inhibit thyroxine synthesis; MMC could affect only stimulatively the pituitary-adrenal axis and PRL synthesis/release, the primary action site for which may be the CNS; and the effects of the pituitary-gonadal axis were inconsistent and, therefore, this axis seems to be relatively resistant to MMC. On the other hand, the responses of PRL and TSH to TRH loading, which were examined for both groups in Exp. 3, suggested that MMC could not affect the metabolizing activity for serum PRL and TSH. The hormone levels of the MMC group enhanced by TRH recovered very rapidly as in the control group. Thus, these acute and reversible endocrine effects seem to indicate relatively earlier development of possible chronic and irreversible effects on the endocrine functions when exposed to methylmercury chronically, and these should be examined further. PMID- 3030713 TI - Diurnal rhythms of proopiomelanocortin-derived N-terminal peptide, beta lipotropin, beta-endorphin and adrenocorticotropin in normal subjects and in patients with Addison's disease and Cushing's disease. AB - In order to clarify the diurnal pattern of secretion of plasma immunoreactive (IR) proopiomelanocortin (POMC)-derived peptides, IR-N-terminal peptide (Nt), IR beta-endorphin (Ep), IR-beta-lipotropin (LPH), and IR-ACTH (ACTH) in normal subjects and in patients with Addison's disease and Cushing's disease, we measured these 4 peptides in the same plasma obtained at 0900 h and then every three hours until 0600 h at the next day. All four peptides showed diurnal rhythms with the peaks at 0600 h, and the nadirs of ACTH, LPH, Ep and Nt were at 0000 h, 0000 h, 1800 h and 0300, respectively in normal subjects. In patients with Addison's disease, these four peptides also showed diurnal rhythms with the peaks at 0600 h for ACTH and Ep and at 0900 h for LPH and Nt, and the nadirs at 2100 h for ACTH and Ep and at 0000 h for LPH and Nt. The molar ratios of Ep/ACTH, LPH/ACTH and Nt/ACTH in plasma also presented diurnal variations in normal subjects and in patients with Addison's disease. On the other hand, in patients with Cushing's disease, ACTH, LPH and Nt showed no rhythmicity or change in molar ratios of Ep/ACTH, LPH/ACTH or Nt/ACTH. Only Ep showed diurnal variation. The molar ratios of Ep/ACTH, LPH/ACTH and Nt/ACTH in patients with Cushing's disease were significantly higher than those in normal subjects and in patients with Addison's disease at 0000 h.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3030714 TI - Serum relaxin in patients with invasive mole, choriocarcinoma and persistent trophoblastic disease. AB - Human chorionic gonadotropin (hCG) is considered to be one of the factors which regulate relaxin secretion in humans. Serum immunoreactive relaxin levels are increased and are detectable by radioimmunoassay both in normal and molar pregnancy. Circulating hCG levels are increased in trophoblastic disease. In the present study, relaxin and hCG levels were sequentially measured in patients with invasive mole, choriocarcinoma and persistent trophoblastic disease. Serum relaxin levels were detectable by radioimmunoassay in these patients before treatment, though they were significantly lower than in normal pregnancy. The corpus luteum of pregnancy is the main source of circulating relaxin in normal pregnancy. The existence of a corpus luteum was confirmed in the 2 patients who underwent laparotomy. Consequently, the corpus luteum may also be the main source of circulating relaxin in trophoblastic disease. Parallel changes in hCG and relaxin levels were observed during the courses of trophoblastic disease. The finding suggests that relaxin secretion is dependent on hCG stimulation in trophoblastic disease in the presence of corpus luteum. PMID- 3030715 TI - Small-cell carcinoma of the esophagus: a case treated by chemotherapy. AB - A patient with small-cell carcinoma of the esophagus treated with cyclophosphamide and vincristine is described. Within 2 months after the beginning of therapy, two coexisting tumors had disappeared and the main primary lesion and metastases in the liver and lymph nodes were markedly reduced in size. The patient relapsed 5 months after the start of therapy and died with widespread metastases. Autopsy revealed that the patient was in partial remission. PMID- 3030716 TI - Hepatocellular carcinoma presenting with bleeding due to duodenal perforation by the tumor. AB - A 70-year-old man was admitted with upper GI bleeding. Endoscopy showed a large duodenal bleeding mass. The aspect of the tumor was macroscopically similar to the liver surface. The diagnosis of hepatocellular carcinoma was made by aspiration cytology guided by abdominal computed tomographic scan. The postmortem examination revealed an extensive primary hepatic tumor which caused an ulcer in the first part of the duodenum and protruded into the lumen. This unusual presentation of a hepatocellular carcinoma has not been previously reported. PMID- 3030717 TI - A case of laparoscopically detected cholangiocarcinoma of a few millimeters in size, with portal fibrosis. AB - A 54-year-old male with no subjective complaints was admitted to our hospital with fatty liver and low echoic lesion of 3 X 4 cm, thereafter revealed to be a hemangioma. A white lesion of few millimeters in size was detected in the left lobe of the liver at laparoscopic examination, and histologically diagnosed in an aimed biopsy as cholangiocarcinoma. Another 5 lesions (three white spots, a white longitudinal protrusion and a white lesion with a lacy pattern) suggestive of malignancy were also selectively biopsied, but all proved histologically negative. Microwave coagulation was applied to all biopsied sites as anti-cancer therapy. PMID- 3030718 TI - Control of steroid synthesis in adrenal fasciculata cells. AB - The control of steroid synthesis in adrenal fasciculata cells is considered in terms of two types of control by ACTH: control of cholesterol availability to inner mitochondrial cytochrome P-450scc. This process controls total steroid synthesis and is rapidly activated by ACTH. The several steps in cholesterol transfer are examined. partitioning of metabolism by means of competition between enzymes for limiting amounts of steroid intermediates. Changes in such competition determine the ratio of steroid products from the adrenal cells. Such changes typically are a slower response to ACTH. A critical aspect of such competition is the modulation of multiple activity P-450 cytochromes: P-450(17 alpha) (17 alpha-hydroxylation and 17,20 lyase) and P-450(11 beta) (11 beta- and 18-oxidases). Factors such as substrate binding, electron transfer steps and lipid environment are considered in addition to new enzyme synthesis. The ACTH stimulation of steroid synthesis in bovine adrenal cell primary cultures is examined as a model for both types of regulation. PMID- 3030719 TI - Sterol carrier protein2: further evidence for its role in adrenal steroidogenesis. AB - Homogeneous rat liver sterol carrier protein (SCP2) has been implicated in adrenal steroidogenesis by studies utilizing as a model system various sub cellular fractions of rat adrenals. Levels of SCP2 were measured in rat adrenal subcellular fractions and various rat tissues using a highly sensitive radioimmunoassay. The levels of SCP2 in various tissues correlate well with the capacity of each tissue to either synthesize or metabolize cholesterol. The high level of SCP2 in adrenal mitochondria (46% of total tissue SCP2) is consistent with its proposed role of enhancing transfer of cholesterol from the outer to the inner mitochondrial membrane. Neither ACTH nor cycloheximide treatment of rats had a significant effect on SCP2 levels or distribution in the adrenal subcellular fractions. Western blot analysis of adrenal subcellular fractions indicates the presence of a protein of identical molecular weight and at least similar antigenicity as homogeneous rat liver SCP2. In the present studies, intact dispersed rat adrenal fasciculata cells fused with liposomal encapsulated anti-SCP2 IgG showed a 40-65% reduction in their ability to produce corticosterone when stimulated with ACTH. The steroidogenic competence of these anti-SCP2 IgG treated cells can be restored by treatment of the cells with liposomal encapsulated SCP2 prior to ACTH stimulation. These findings provide direct evidence for the involvement of SCP2 in ACTH stimulated steroidogenesis in rat adrenocortical cells, and suggests that SCP2 may not be the putative high turnover "labile protein" involved in acute steroidogenesis. PMID- 3030720 TI - Acute cAMP stimulation in Leydig cells: rapid accumulation of a protein similar to that detected in adrenal cortex and corpus luteum. AB - Addition of cAMP (as the dibutyryl compound) to a primary culture of mouse Leydig cells caused the accumulation of a protein, i(b), with the same dependence on cAMP concentration as the increase in testosterone synthesis. Stimulation of both protein i(b) and testosterone production were inhibited by cycloheximide. Additionally, cAMP caused repression of synthesis of another protein, p(b), with the same approximate molecular weight (28,000 daltons) as i(b), but more basic isoelectric point. This behavior resembles an event which has been documented in the adrenal cortex (Krueger, R.J., and Orme-Johnson, N.R., J. Biol. Chem., 258, 10159-10167, 1983) and corpus luteum (Pon, L.A., and Orme-Johnson, N.R., J. Biol. Chem., 261, 6594-6599, 1986). The discovery of these proteins in a third steroid producing cell type and the close correlation between conditions causing increased steroid synthesis and increased i(b) production is further indication that protein i(b) may be an intermediary in peptide hormone or cAMP control of steroid hormone biosynthesis. PMID- 3030721 TI - The action of phorbol ester on steroidogenesis in cultured human fetal adrenal cells. AB - The potent mitogen and tumor promoter, phorbol 12-myristate 13-acetate (PMA), has a primary action via activation of calcium-dependent protein kinase C. The treatment of monolayer cultures of human fetal adrenal neocortex (HFA) cells with PMA (50-250 nM) stimulated basal dehydroepiandrosterone sulfate (DS) secretion 2 3 fold. ACTH-treated HFA cells secreted amounts of DS and cortisol (F) 10-50 fold greater than basal secretions. PMA (250 nM) addition with ACTH to HFA cells decreased DS and F secretions at least 75% on days 2 and 3 of treatment. Treatment of HFA cells with 4 alpha-phorbol, which does not activate calcium dependent protein kinase C, did not inhibit steroidogenesis. The attenuated rates of steroidogenesis after PMA treatment correlated with the decreased amounts of steroid 11 beta, 17 alpha- and 21-hydroxylase cholesterol side-chain cleavage steroid dehydrogenase and sulfotransferase activities. The decrease of steroid 17 alpha-hydroxylase activity correlated with the decreased amount of cytochrome P 450(17) alpha as determined after protein immunoblotting of NaDodSO4 cell lysates. After PMA treatment the ACTH-promoted increases of hydroxysteroid sulfotransferase and dehydrogenase activities of HFA cells were suppressed. PMA (50 nM) inhibited cAMP accumulation in ACTH-treated HFA cells, while 4 alpha phorbol had no effect. Importantly, dibutyryl cAMP (0.2 mM) treatment of HFA cells did not reverse phorbol ester-promoted attenuation of steroidogenesis. We conclude that, in the presence of ACTH, phorbol ester chronically inhibits both cAMP synthesis and cAMP-dependent protein kinase action with resultant decreased steroidogenic enzyme synthesis and steroid production. This may be a consequence of activation, migration and a slow degradation of protein kinase C activity. These multifaceted actions of phorbol ester and associated protein kinase C activation may have critical effects on the ontogeny of fetal adrenal function. PMID- 3030722 TI - Corticotropin specifically stimulates the uptake and intracellular movements of sterol carrier protein in adrenals. AB - Sterol carrier protein (SCP), also known as liver fatty acid binding protein, is a major adrenal protein localized in the cytosol and inner mitochondrial membrane. SCP is synthesized in liver and intestine and then rapidly secreted into blood, where it associates primarily with the high density lipoprotein fraction. Studies using steroidogenesis inhibitors showed that corticotropin has a specific effect on the uptake of SCP and its movement with cholesterol to the inner mitochondrial membrane. Thus, SCP appears to be intimately involved in the mechanism(s) of adrenal steroidogenesis from cholesterol. PMID- 3030723 TI - Molecular genetics and the characterization of steroid 21-hydroxylase deficiency. AB - Classical 21-hydroxylase deficiency congenital adrenal hyperplasia is a monogenic autosomal recessive disorder that has been conclusively shown by family HLA typing studies to be in close genetic linkage with the human major histocompatibility complex. More recently recognized is the nonclassical disorder, an attenuated form of 21-hydroxylase deficiency characterized variably by late onset or absence of symptoms. Certain of the mild 21-hydroxylase deficiency allotypes involved in the nonclassical disorder have also been shown to be genetically linked with HLA, exhibiting distinct (B and B,DR) antigen associations. The nonclassical disorder is now also known to result from different genotypes: two mild 21-hydroxylase defects in conjunction, or a mild defect occurring with a sever (classical) defect. Restriction mapping and hybridization analysis have located two highly homologous base sequences, one structural gene coding for 21-hydroxylase and one pseudogene, in the Class III region of the MHC in tandem with the A and B genes for C4, the fourth component of complement. Current work documenting and characterizing gene abnormalities, as well as elucidating the molecular genetic basis of the mutations that have arisen, is aimed at developing better cDNA probes for prenatal diagnosis by amniocentesis and chorionic villus biopsy. In addition, because of the close association of the C4 and 21-hydroxylase genes, coordination of data on C4 variants and null alleles with altered 21-hydroxylase activity is improving understanding of the genetic mechanisms generating disease alleles of this enzyme crucial for normal endocrine function. PMID- 3030724 TI - Calcium and the control of renin secretion. PMID- 3030725 TI - Effect of induced alkalosis on physical work capacity during arm and leg exercise. PMID- 3030726 TI - Reconstitution of the fusogenic activity of vesicular stomatitis virus. AB - Enveloped virus glycoproteins exhibit membrane fusion activity. We have analysed whether the G protein of vesicular stomatitis virus, reconstituted into liposomes, is able to fuse nucleated cells in a pH-dependent fashion. Proteoliposomes produced by octylglucoside dialysis did not exhibit cell fusion activity of the G protein. However, by making use of n-dodecyl octaethylene monoether (C12E8) as the solubilizing agent and by removal of the detergent in two steps, we were able to produce fusogenic G protein liposomes. These G protein liposomes fuse to the BHK-21 cell surface at pH 5.7-6.0 with an efficiency of fusion comparable with that of the parent virus. Physical and chemical analysis revealed that the fusogenic liposomes exhibited a protein to lipid weight ratio of 0.67 and showed an average diameter of 130 nm. PMID- 3030728 TI - Rearrangement of immunoglobulin heavy chain genes in human T leukaemic cells shows preferential utilization of the D segment (DQ52) nearest to the J region. AB - The DNA rearrangements leading to the assembly of genes coding for the immunoglobulin heavy chain (IgH) in B cells and the T cell receptor for antigen in T cells are not completely lineage specific. This probably reflects the use of a common recombinase by IgH and the T cell receptor. This paper describes novel observations on the nature of these cross-lineage rearrangements. A high proportion (though not all) IgH rearrangements in human T leukaemic cells involve the D segment nearest to the J region (DQ52). This same D segment is not involved in B cell IgH rearrangements with one important exception, namely a proportion of B cell leukaemic clones with the most primitive B cell precursor phenotype. These observations have potentially important implications for early lymphoid cell differentiation and in particular support the idea that the 3' D plus J region might lie within a limited window of accessibility of the IgH gene in precursor lymphocytes. PMID- 3030727 TI - Regulation of protein kinase activities in PC12 pheochromocytoma cells. AB - Stimulation of serine protein kinase activity (referred to as S6 kinase) occurs within minutes of addition of nerve growth factor (NGF) to PC12 rat pheochromocytoma cells. This enzyme activity is not related to the cAMP-dependent protein kinase (protein kinase A) or the Ca2+- and phospholipid-dependent protein kinase (protein kinase C), two other protein kinases potentially involved in signal transduction. Two peaks of NGF-stimulated S6 phosphotransferase activity are observed upon ion exchange chromatography; one that comigrates with the serine kinase previously described in chicken embryo fibroblasts and another with distinct elution properties. Several other factors are also found to regulate S6 phosphotransferase activity in PC12 cells including epidermal growth factor, insulin, and phorbol myristate acetate. Dibutyryl cAMP stimulates S6 phosphotransferase activity; however, this activity is strongly inhibited by the protein kinase A heat stable inhibitor. At least two mechanisms exist through which the NGF-stimulated S6 kinase activity can be regulated, one that apparently can use protein kinase C whereas the other(s) does not. The potential roles of these protein kinase activities in signal transduction and regulation of cell growth and differentiation is discussed. PMID- 3030729 TI - The complete sequence and structural analysis of human apolipoprotein B-100: relationship between apoB-100 and apoB-48 forms. AB - We have isolated and sequenced overlapping cDNA clones covering the entire sequence of human apolipoprotein B-100 (apoB-100). DNA sequence analysis and determination of the mRNA transcription initiation site by S1 nuclease mapping showed that the apoB mRNA consists of 14,112 nucleotides including the 5' and 3' untranslated regions which are 128 and 301 nucleotides respectively. The DNA derived protein sequence shows that apoB-100 is 513,000 daltons and contains 4560 amino acids including a 24-amino-acid-long signal peptide. The mol. wt of apoB 100 implies that there is one apoB molecule per LDL particle. Computer analysis of the predicted secondary structure of the protein showed that some of the potential alpha helical and beta sheet structures are amphipathic, whereas others have non-amphipathic neutral to apolar character. These latter regions may contribute to the formation of the lipid-binding domains of apoB-100. The protein contains 25 cysteines and 20 potential N-glycosylation sites. The majority of cysteines are distributed in the amino terminal portion of the protein. Four of the potential glycosylation sites are in predicted beta turn structures and may represent true glycosylation positions. ApoB lacks the tandem repeats which are characteristic of other apolipoproteins. The mean hydrophobicity the mean value of H1 and helical hydrophobic moment the mean value of microH profiles of apoB showed the presence of several potential helical regions with strong polar character and high hydrophobic moment. The region with the highest hydrophobic moment, between amino acid residues 3352 and 3369, contains five closely spaced, positively charged residues, and has sequence homology to the LDL receptor binding site of apoE. This region is flanked by three neighbouring regions with positively charged amino acids and high hydrophobic moment that are located between residues 3174 and 3681. One or more of these closely spaced apoB sequences may be involved in the formation of the LDL receptor-binding domain of apoB-100. Blotting analysis of intestinal RNA and hybridization of the blots with carboxy apoB cDNA probes produced a single 15-kb hybridization band whereas hybridization with amino terminal probes produced two hybridization bands of 15 and 8 kb. Our data indicate that both forms of apoB mRNA contain common sequences which extend from the amino terminal of apoB-100 to the vicinity of nucleotide residue 6300. These two messages may have resulted from differential splicing of the same primary apoB mRNA transcript. PMID- 3030730 TI - Involvement of finger domain and kringle 2 domain of tissue-type plasminogen activator in fibrin binding and stimulation of activity by fibrin. AB - Human tissue-type plasminogen activator (t-PA) catalyses the conversion of inactive plasminogen into active plasmin, the main fibrinolytic enzyme. This process is confined to the fibrin surface by specific binding of t-PA to fibrin and stimulation of its activity by fibrin. Tissue-type plasminogen activator contains five domains designated finger, growth factor, kringle 1, kringle 2 and protease. The involvement of the domains in fibrin specificity was investigated with a set of variant proteins lacking one or more domains. Variant proteins were produced by expression in Chinese hamster ovary cells of plasmids containing part of the coding sequence for the activator. It was found that kringle 2 domain only is involved in stimulation of activity by fibrin. In the absence of plasminogen and at low concentration of fibrin, binding of t-PA is mainly due to the finger domain, while at high fibrin concentrations also kringle 2 is involved in fibrin binding. In the presence of plasminogen, fibrin binding of the kringle 2 region of t-PA also becomes important at low fibrin concentrations. PMID- 3030731 TI - Sites hypersensitive to, and protected from, nuclease digestion in the regulatory region of wild-type and mutant polyoma chromatin. AB - It has been shown that the untranscribed regulatory region of polyoma virus (Py) is hypersensitive (Hs) to DNase I treatment, and that this hypersensitivity is located in two areas which correspond to the A and B domains of the enhancer. We mapped the DNase I hypersensitive sites in the Py regulatory region of wild-type (PyA2) and of mutants, selected in neuroblastoma cells (PyNB), which are characterized by an extensive duplication involving the A domain, with or without deletion of the B domain. The experiments were performed in both a permissive host (3T6 mouse fibroblasts) and in a restrictive host (41A3 mouse neuroblasts). No significant differences were observed between the two hosts. Our results show that four sites, in addition to the ones already described, can be identified in the wild-type A2 strain. These newly identified sites coincide with the domains of the enhancer region as they have recently been established. In PyNB mutants duplications and deletions are generally correlated to the gain or loss of the corresponding hypersensitive sites. However, a new site is formed in one of the duplicated sequences, even if no corresponding hypersensitive site is present in the other identical sequence. A region protected from DNase I digestion occurs in the PyNB mutants which corresponds to the junction of the duplication which is absent in the wild-type strain. In this region, as a consequence of the rearrangement, a GGCGGG motif which is very similar to the one (GGGCGG) present at the binding sites of the cellular regulatory protein SP1, is found.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3030733 TI - The sex-determining gene tra of Drosophila: molecular cloning and transformation studies. AB - In Drosophila, the primary signal for sex determination is given by the ratio of X chromosomes to sets of autosomes (X:A). The primary signal is read by a key gene (Sxl) and transmitted down to the differentiation genes by the subordinate control genes tra, tra-2, ix and dsx. Mutations in tra transform chromosomal females (X/X; tra/tra) into sterile males (pseudomales). We have cloned the tra region by microdissection and chromosomal walking. We identified the gene using deficiency breakpoints, DNA aberrations in three different alleles of tra and by P-mediated transformation. A 3.8-kb fragment perfectly rescued the mutant phenotype of X/X; tra/tra flies, showing that it contained all the necessary information to restore female-specific functions in the mutant flies. We present evidence that most of the function of tra can be provided by a subsegment of 2 kb that is differentially transcribed or processed in males and females. PMID- 3030732 TI - The first exon of the c-myc proto-oncogene contains a novel positive control element. AB - We have identified a positive modulator within the c-myc first exon downstream of the gene's transcription initiation sites, P1 and P2. We introduced myc-CAT (chloramphenicol acetyltransferase) hybrid genes into three cell lines (BJAB, COS and HeLa) and measured their expression by either CAT enzymatic activity, S1 nuclease protection or by a nuclear 'run-on' transcription assay. Removal of 46 bp from the 3' end of the first exon results in a decrease of myc-CAT expression and P2 activity. A 438-bp exon 1 segment, lacking the normal myc promoters, efficiently drives the expression of SV40 early promoters. We find that this first exon segment efficiently functions as a positive modulator only in its sense orientation, 3' of a nearby promoter. The positive effects of the myc first exon and the SV40 enhancer are complementary. PMID- 3030735 TI - Transposition of mini-Mu containing only one of the ends of bacteriophage Mu. AB - Transposition of mini-Mu containing only one of the ends of bacteriophage Mu was studied. Transposition was observed when plasmids containing this mini-Mu were used as the donor in a mating-out assay with the F factor POX38, which lacks all known transposable elements, as the target. Analysis of the plasmids isolated from the recipient strain showed that in most cases the mini-Mu containing plasmid and POX38 were fused and that a part of the plasmid had been duplicated, indicating the formation of co-integrates. To obtain the DNA sequences of the junctions between donor and recipient DNA, an F factor was constructed that made it possible to analyse these junctions directly. The results showed that several sequences can be used as an alternative end in transposition and that these alternative ends show homology with the consensus for a strong A binding site. Moreover, the first base pair at the junction was always a (TA) base pair. This base pair is situated at exactly the same position with respect to the sequence which has homology with the consensus for a strong A binding site as in the ends of Mu. PMID- 3030734 TI - At least two nuclear gene products are specifically required for translation of a single yeast mitochondrial mRNA. AB - Mitochondrial translation of the oxi2 mRNA, encoding yeast cytochrome c oxidase subunit III (coxIII), has previously been shown to specifically require the mitochondrially located protein product of the nuclear gene PET494. We show here that this specific translational activation involves at least one other newly identified gene termed PET54. Mutations in PET54 cause an absence of the coxIII protein despite the presence of normal levels of its mRNA. pet494 mutations are known to be suppressible by mitochondrial gene rearrangements that replace the normal 5'-untranslated leader of the oxi2 mRNA with the leaders of other mitochondrial mRNAs. In this study we show that pet54, pet494 double mutants are suppressed by the same mitochondrial gene rearrangements, showing that the PET54 product is specifically required, in addition to the PET494 protein, for translation of the oxi2 mRNA. Since, as we show here, PET54 is not an activator of PET494 gene expression, our results suggest that the products of both of these genes may act together to stimulate coxIII translation. PMID- 3030736 TI - Illegitimate recombination occurs between the replication origin of the plasmid pC194 and a progressing replication fork. AB - Hybrids between plasmids pC194, pBR322 and the bacteriophage f1 undergo deletions in Escherichia coli. The deletions end most often between nucleotides 1445 and 1446 of pC194. That site probably corresponds to a nick in the replication origin of this plasmid. The localization of the other deletion end appears to be determined by the position of the f1 replication fork. Two models accounting for these data are discussed. PMID- 3030737 TI - Inducible cephalosporinase production in clinical isolates of Enterobacter cloacae is controlled by a regulatory gene that has been deleted from Escherichia coli. AB - Cephalosporin hyper-resistant Enterobacter cloacae strains are isolated with increasing frequency from hospital infections. Resistance is principally due to the chromosomal ampC gene encoding a cephalosporinase. In contrast to Escherichia coli which expresses ampC constitutively from a promoter located in the upstream frdD gene, E. cloacae displays inducible ampC expression. By cloning the ampC gene it was shown that a linked genetic locus, ampR, mediated the induction by beta-lactams. In the absence of the antibiotic the 30,500 dalton AmpR protein represses ampC expression. The ampR gene shows a highly compact arrangement and is situated between the divergently expressed ampC gene and the frd operon from which it is separated by a bifunctional transcription terminator. The promoters for ampR and ampC substantially overlap and mRNA analyses showed that on induction transcription from the ampC promoter increased greatly whereas that from ampR did not. Two regions of sequence homology flank the ampR gene and it is proposed that a homologous recombination event between these in an ancestral enteric bacterium may have led to the deletion of ampR from the E. coli genome. PMID- 3030738 TI - The origin and use of the terms competitive and non-competitive in interactions among chemical substances in biological systems. AB - The terms competition and competitive were in use for appropriate types of interaction in human and animal behaviour from the seventeenth century. In the nineteenth and early twentieth centuries they reached more technical uses in biology, especially in darwinian studies; and in chemistry in describing competing reactions, surface phenomena and the influence of substituent groupings in reactant molecules. Use of competitive and non-competitive to describe enzyme inhibitors had a specific beginning when J. B. S. Haldane (following premonitory work of others) applied the terms in 1927 and 1930 to types of inhibition already differentiated by Michaelis and co-workers. The theoretical background in kinetics and stereochemistry so acquired gave a firmness to the application of the terms in biochemistry. The first examples concerned glycosidases, especially beta-D-fructofuranosidase or invertase, and interactions of carbon monoxide and oxygen at iron-porphyrin systems. They were thus of interest in toxicology and in enzyme and carrier studies. The sphere of application of the biochemically defined terms expanded greatly when, following investigation of sulphonamide action, it was realized that concepts of enzyme inhibition by structurally related compounds offered a route to understanding the action of existing medicaments and to the production of new ones. Ideas and terminology based on competitive and non-competitive enzyme inhibition and receptor occupancy have subsequently been applied in many ways. Examples include application to the analysis of feedback inhibition and other processes of metabolic control; to receptor relationships among neurotransmitters and medicaments; and to understanding interactions at sensory receptors. PMID- 3030739 TI - Primary structure of the Streptomyces R61 extracellular DD-peptidase. 2. Amino acid sequence data. AB - In order to confirm the Streptomyces codon usage, the Streptomyces R61 DD peptidase was fragmented by cyanogen bromide cleavage of the carboxymethylated protein, trypsin digestion of the carboxymethylated protein and trypsin digestion of the protein treated with beta-iodopenicillinate and endoxo-delta 4 tetrahydrophthalic acid. The isolated peptides, which altogether represented more than 50% of the polypeptide chain, were sequenced. The data thus obtained were in excellent agreement with the primary structure of the protein as deduced from the nucleotide sequence of the cloned gene. Though a weak acylating agent, beta iodopenicillanate reacted selectively with the active site of the DD-peptidase and formed an adduct which mas much more stable than that formed with benzylpenicillin, thus facilitating the isolation and characterization of the active-site peptide. PMID- 3030740 TI - Characterization of the cytochrome system of a nitrogen-fixing strain of a sulfate-reducing bacterium: Desulfovibrio desulfuricans strain Berre-Eau. AB - Two c-type cytochromes were purified and characterized by electron paramagnetic resonance (EPR) and nuclear magnetic resonance (NMR) spectroscopic techniques, from the sulfate-reducer nitrogen-fixing organism, Desulfovibrio desulfuricans strain Berre-Eau (NCIB 8387). The purification procedures included several chromatographic steps on alumina, carboxymethylcellulose and gel filtration. A tetrahaem and a monohaem cytochrome were identified. The multihaem cytochrome has visible, EPR and NMR spectra with general properties similar to other low potential bis-histidinyl axially bound haem proteins, belonging to the class of tetrahaem cytochrome c3 isolated from other Desulfovibrio species. The monohaem cytochrome c553 is ascorbate-reducible and its EPR and NMR data are characteristic of a cytochrome with methionine-histidine ligation. Their properties are compared with other homologous proteins isolated from sulfate reducing bacteria. PMID- 3030741 TI - Complementation between LLC-PK1 mutants affected in polypeptide hormone-receptor function. AB - The mutant LLC-PK1 cell lines FIB6 and FIB5/N4 were examined for responsiveness to the polypeptide hormones calcitonin and vasopressin. Both mutants exhibited little or no activation of adenylate cyclase or cAMP-dependent protein kinase (cAMP-PK) in response to calcitonin, but responded to vasopressin. Analysis of calcitonin receptor function demonstrated that both mutants bound less than 9 fmol 125I-labeled salmon calcitonin/mg cellular protein, which was about 1% of parental activity (642 fmol calcitonin bound/mg). Concomitant with reduced calcitonin binding, both mutants exhibited increased vasopressin binding (greater than 272 fmol [[3H]Arg]vasopressin bound/mg) compared to parental (166 fmol bound/mg). The concentration of vasopressin for half-maximal stimulation of adenylate cyclase in both mutants was comparable to that for LLC-PK1 cells (40 pM) and hence the increased binding activity was concluded to be due to increased numbers of functional vasopressin receptors in the mutants. Somatic cell hybrids formed between each mutant and LLC-PK1 cells exhibited normal hormone binding and activation of cAMP-PK in response to both vasopressin and calcitonin. The mutations affecting receptor function in FIB6 and FIB5/N4 were accordingly concluded to be recessive. Somatic cell hybrids between FIB6 and FIB5/N4 showed no complementation of the mutant phenotype, indicating that both cell lines were affected in the same gene. In contrast, somatic cell hybrids between FIB5/N4 and the 'receptorless' mutant M18 (which lacks functional calcitonin and vasopressin receptors) exhibited approximately the same responsiveness to vasopressin and to calcitonin as LLC-PK1. Complementation between two different mutations affecting polypeptide receptor function was thus observed. The results are discussed in terms of a proposed common pathway for processing of calcitonin and vasopressin receptors. PMID- 3030742 TI - 13C-NMR, 1H-NMR and gas-chromatography mass-spectrometry studies of the biosynthesis of 13C-enriched L-lysine by Brevibacterium flavum. AB - The basic metabolic pathways of lysine biosynthesis in Brevibacterium flavum, a strain which excretes excessive amounts of L-lysine, have been followed by using two 13C-labeled precursors. 13C- and 1H-NMR spectroscopies in conjunction with gas chromatography mass spectrometry (GC-MS) have revealed the various metabolic pathways leading to L-[13C]lysine. Discrete metabolic pathways give rise to distinct labeling patterns. L-Lysine resulting from [1-13C]glucose fermentation is relatively specifically labeled: L-[3,5-13C]lysine is the main product. Experimental and theoretical approaches based on the 13C-enrichment values of intracellular glutamate, a major intermediate metabolite, allowed us to assess the relative contribution of the major metabolic pathways forming lysine. The labeling pattern of glutamate reflects the isotope distribution in 2 oxoglutarate. When [2-13C]acetate is used as the sole carbon source in the culture, the energy-producing steps of the Krebs cycle are essential. The higher activity of the Krebs cycle, when endogenous carbohydrates are exhausted from the culture, is indicated by the increased 13C enrichment in C-1 of lysine and reveal a high content of isotopomers of four, five and six 13C atoms in the lysine molecule, pointing out that the four-carbon intermediates of the cycle are being derived from the glyoxylate shunt pathway. Such a phenomenon does not occur in glucose fermentation. GC-MS analyses of 13C enrichments and isotopomer distributions in metabolites and end products are in good agreement with the predicted contribution of each metabolic pathway. This new methodological approach of combined NMR and GC-MS has been demonstrated to be applicable to various other metabolic studies. PMID- 3030743 TI - The relationship between beta-adrenoceptor regulation and beta-adrenergic responsiveness in hepatocytes. Studies on acquisition, desensitization and resensitization of isoproterenol-sensitive adenylate cyclase in primary culture. AB - The role of beta-adrenoceptor regulation in the mechanisms controlling beta adrenergic responsiveness in hepatocytes was explored, using primary monolayer cultures. When plated in vitro, these cells gradually acquire a strong catecholamine-sensitive adenylate cyclase activity and an enhanced ability to bind the beta-adrenoceptor ligand [125I]iodocyanopindolol (125ICYP). Examination of the time course showed that the increase in the number of 125ICYP binding sites was detectable within 1-2 h of culturing and slightly preceded the elevation of isoproterenol-responsive activity. Then the responsiveness rose steeply and between about 5-24 h it closely followed the increase in beta receptor binding. Addition of isoproterenol (10 microM) to cells after 20 h of culturing caused a rapid homologous desensitization of the adenylate cyclase (50% after about 5 min). This was paralleled by a down-regulation of beta adrenoceptors measured both in membrane particles and in total cell lysates. Removal of isoproterenol led to a resensitization of the adenylate cyclase, which was rapid and protein-synthesis-independent after a brief (10-min) desensitization, or slow and cycloheximide-sensitive after prolonged (4-h) exposure to the agonist. In both cases an up-regulation of the 125ICYP binding paralleled the recovery from refractoriness. In contrast, no concurring changes in 125ICYP binding were measured when the beta-adrenoceptor-linked adenylate cyclase activity was enhanced by pretreatment with pertussin toxin (islet activating protein, IAP) or was desensitized by exposure of the cells to glucagon or 8-bromo-cAMP; however, these modulations of the adenylate cyclase were nonselective, since the pretreatments with IAP, glucagon or 8-bromo-cAMP affected both isoproterenol-sensitive and glucagon-sensitive activities. The results suggest that, in hepatocytes, regulation at the beta-adrenoceptor level is a major determinant for both short-term and long-term selective changes of the beta adrenergic responsiveness. PMID- 3030744 TI - Purification, subunit structure and properties of a high-molecular-mass protein phosphatase capable of dephosphorylating mRNP of the brine shrimp Artemia sp. AB - A phosphoprotein phosphatase active towards casein, phosphorylase a and mRNP proteins has been detected in the cytosol of cryptobiotic gastrulae of Artemia sp. This phosphatase has a relative molecular mass (Mr) of 225,000 as measured by gel filtration on Sephadex G-200 and has been purified to near homogeneity by ion exchange chromatography on different DEAE-substituted matrices, affinity chromatography on polylysine-agarose, histone-Sepharose 4B and protamine-agarose, hydrophobic chromatography on phenyl-Sepharose 4B and gel filtration on Sephadex G-200. Sodium dodecyl sulphate gel electrophoresis of the final purification step revealed that the enzyme contains two types of subunits, alpha and beta, with Mr of 40,000 and 75,000, respectively. These values, in conjunction with the native Mr and the molar ratios of the subunits estimated by densitometric analysis of the gel, suggested that the subunit composition of the enzyme is alpha 2 beta 2. When treated with 1.7% (v/v) 2-mercaptoethanol at -20 degrees C or with ethanol, the enzyme released the catalytic alpha subunit of Mr 40,000. The protein phosphatase was activated by basic proteins e.g. protamine (A 0.5 = 1 microM), histone H1 (A 0.5 = 1.6 microM) and polylysine (A 0.5 = 0.2 microM) and inhibited by ATP (I 0.5 = 12 microM), NaF (I 0.5 = 3.1 mM) and pyrophosphate (I 0.5 = 0.6 mM). The enzyme is a polycation-stimulated protein phosphatase. Purified mRNP proteins, phosphorylated by the mRNP-associated casein kinase type II, are among the substrates used by the enzyme. The function of reversible phosphorylation dephosphorylation of mRNP as a regulatory mechanism in mRNP metabolism is discussed. PMID- 3030745 TI - Disassembly and domain structure of the proteins in the signal-recognition particle. AB - The signal-recognition particle (SRP) is a ribonucleoprotein (RNP) complex consisting of six different polypeptide chains and a 7SL RNA. It participates in initiating the translocation of proteins across the membrane of the endoplasmic reticulum. SRP was disassembled in 2 M KCl into three components, one RNP composed of 7SL RNA and the 54-kDa and 19-kDa proteins, and two heterodimers consisting of the 72/68-kDa and the 14/9-kDa proteins respectively. The 54-kDa protein could be released from the RNP subparticle by chromatography on DEAE Sepharose in Mg2+-depleted buffer, while the 19-kDa protein remained bound to the 7SL RNA. The domain structure of SRP proteins was probed by using mild elastase treatment and protein-specific antibodies. It was found that the 72, 68, 54 and 19-kDa SRP proteins were proteolytically processed in distinct steps. Most remarkably a protein fragment of 55-kDa, generated from the 72-kDa SRP protein, and a 35-kDa fragment from the 54-kDa SRP protein were both released from the RNP particle. Fragments generated from the 68-kDa protein and detectable with the anti-(68-kDa protein) antibody remained associated with the RNP particle. Cleavage of the SRP proteins by elastase at 2.5 micrograms/ml resulted in partial loss of activity, while 10 micrograms/ml caused complete inactivation of the particle. Neither the elongation arrest of IgG light chain nor its translocation across SRP-depleted microsomal membranes was promoted. The implications of these results on the possible interaction between the SRP subunits are discussed. PMID- 3030746 TI - Site-directed mutagenesis of the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase from Anacystis nidulans. AB - Using oligonucleotide-directed mutagenesis of the gene encoding the small subunit (rbcS) from Anacystis nidulans mutant enzymes have been generated with either Trp 54 of the small subunit replaced by a Phe residue, or with Trp-57 replaced by a Phe residue, whereas both Trp-54 and Trp-57 have been replaced by Phe residues in a double mutant. Trp-54 and Trp-57 are conserved in all amino acid sequences or the small subunit (S) that are known at present. The wild-type and mutant forms of Rubisco have all been purified to homogeneity. The wild-type enzyme, purified from Escherichia coli is indistinguishable from enzyme similarly purified from A. nidulans in subunit composition, subunit molecular mass and kinetic parameters (Vmax CO2 = 2.9 U/mg, Km CO2 = 155 microM). The single Trp mutants are indistinguishable from the wild-type enzyme by criteria (a) and (b). However, whereas, Km CO2 is also unchanged, Vmax CO2 is 2.5-fold smaller than the value for the wild-type enzyme for both mutants, demonstrating for the first time that single amino acid replacements in the non-catalytic small subunit influence the catalytic rate of the enzyme. The specificity factor tau, which measures the partitioning of the active site between the carboxylase and oxygenase reactions, was found to be invariant. Since tau is not affected by these mutations we conclude that S is an activating not a regulating subunit. PMID- 3030747 TI - The primary structure of cytochrome c1 from Neurospora crassa. AB - The primary structure of the cytochrome c1 subunit of ubiquinol-cytochrome-c reductase from mitochondria of Neurospora crassa was determined by sequencing the cDNA of a bank cloned in Escherichia coli. From the coding region the sequence of 332 amino acids, corresponding to the molecular mass of 36,496 Da, was derived for the precursor protein. The mature protein, the N terminus of which was previously sequenced [Tsugita et al. (1979) in Cytochrome oxidase (King, T. E. et al., eds) pp. 67-77, Elsevier, New York], consists of 262 amino acids and has the molecular mass of 29,908 Da including the heme. The sequence contains an N terminal hydrophilic part of 211 residues, which carries the heme, a hydrophobic stretch of 15 residues, which is assumed to anchor the protein to the membrane, and a C-terminal hydrophilic part of 36 residues. The N-terminal presequence of 70 amino acids contains 9 positive charges but only 1 negative charge and is characterized by a stretch of 20 uncharged residues. PMID- 3030748 TI - The effect of cytochrome c oxidase on lipid polymorphism of model membranes containing cardiolipin. AB - The effect of cytochrome c oxidase incorporation on the lipid polymorphism of the cardiolipin-Ca2+ system was investigated by 31P NMR and freeze-fracture electron microscopy. The integral membrane protein has a stabilizing effect on the bilayer organization of cardiolipin, in that it inhibits the Ca2+-induced HII phase formation of this lipid for Ca2+/cardiolipin molar ratios of 1-10. At a Ca2+/cardiolipin molar ratio of 25, about 80% of the lipid is organized in the HII phase and a structural phase separation occurs between the cardiolipin-Ca2+ complex organized in the hexagonal HII phase without protein and bilayer structures with incorporated protein. PMID- 3030749 TI - New pore protein produced in cells lysogenic for Escherichia coli phage HK253hrk. AB - Outer membrane pore protein OmpC was identified as the receptor for the temperate Escherichia coli phage HK253hrk. The part of OmpC protein recognized by the phage was identified by using hybrid proteins in which parts of OmpC protein are replaced by the corresponding parts of the related PhoE protein. In contrast to other OmpC-specific phages, HK253hrk recognizes a part of OmpC within the C terminal 50 amino acids of the protein. E. coli strains lysogenic for HK253hrk produce reduced amounts of OmpC protein, and produce a new pore protein instead. Expression of this new protein was temperature-dependent, i.e. low at 30 degrees C. The functioning of this new pore protein was characterized both in vivo by studying the uptake of beta-lactam antibodies and in vitro after reconstitution of the protein in black lipid films. Its effective pore size was larger than that of the OmpF pores of E. coli B. The new porin appears to be cation-selective. A comparison with the selectivity of the known OmpC and OmpF pores of E. coli showed that the new pore has a higher selectivity than OmpF but is less selective than OmpC. The new pore protein appears to function in E. coli K12 lysogens as the receptor for the phages HK187, HK189 and HK332. PMID- 3030750 TI - Import of cytochrome c into mitochondria. Cytochrome c heme lyase. AB - The import of cytochrome c into mitochondria can be resolved into a number of discrete steps. Here we report on the covalent attachment of heme to apocytochrome c by the enzyme cytochrome c heme lyase in mitochondria from Neurospora crassa. A new method was developed to measure directly the linkage of heme to apocytochrome c. This method is independent of conformational changes in the protein accompanying heme attachment. Tryptic peptides of [35S]cysteine labelled apocytochrome c, and of enzymatically formed holocytochrome c, were resolved by reverse-phase HPLC. The cysteine-containing peptide to which heme was attached eluted later than the corresponding peptide from apocytochrome c and could be quantified by counting 35S radioactivity as a measure of holocytochrome c formation. Using this procedure, the covalent attachment of heme to apocytochrome c, which is dependent on the enzyme cytochrome c heme lyase, could be measured. Activity required heme (as hemin) and could be reversibly inhibited by the analogue deuterohemin. Holocytochrome c formation was stimulated 5--10 fold by NADH greater than NADPH greater than glutathione and was independent of a potential across the inner mitochondrial membrane. NADH was not required for the binding of apocytochrome c to mitochondria and was not involved in the reduction of the cysteine thiols prior to heme attachment. Holocytochrome c formation was also dependent on a cytosolic factor that was necessary for the heme attaching step of cytochrome c import. The factor was a heat-stable, protease-insensitive, low-molecular-mass component of unknown function. Cytochrome c heme lyase appeared to be a soluble protein located in the mitochondrial intermembrane space and was distinct from the previously identified apocytochrome c binding protein having a similar location. A model is presented in which the covalent attachment of heme by cytochrome c heme lyase also plays an essential role in the import pathway of cytochrome c. PMID- 3030751 TI - Inhibition of proto-oncogene c-fos transcription by inhibitors of protein kinase C and ion transport. AB - Both 4 beta-phorbol 12,13-didecanoate (PDD) and calcium ionophore A23187 induced c-fos mRNA accumulation with similar kinetics in human monocyte-like cells (U937). Their effects were additive. Cells pretreated with PDD were not induced to accumulate c-fos mRNA by PDD but remained responsive to the induction by A23187. Similarly cells pretreated with A23187 responded to PDD but not to A23187. Nuclear run-off transcription assay indicated that increase in the c-fos mRNA level after the second inducer treatment corresponded to transcriptional activation of the c-fos gene. The accumulation of c-fos mRNA and the transcriptional activation of c-fos induced by PDD or A23187 were inhibited by the protein kinase inhibitor H-7 and by quinidine and amiloride. The induction by A23187, but not that by PDD, was also inhibited by 4-aminopyridine, or tetraethylammonium. From these results, it is proposed that activation of protein kinase C and Na+ or K+ transport are required for c-fos induction caused by PDD and A23187. PMID- 3030752 TI - Purification, crystallisation and preliminary X-ray diffraction characterisation of methanol dehydrogenase from Methylosinus trichosporium OB3b. AB - Methanol dehydrogenase was purified from the obligate methanotroph, Methylosinus trichosporium OB3b, in two steps from disrupted biomass by aqueous two-phase partition and ion-exchange chromatography. Copartitioning of a cytochrome c was dependent upon the pH at which aqueous partition was carried out. The native enzyme has a Mr of 120,000, as determined by gel filtration chromatography, and consists of two identical subunits. The purified enzyme contained four electrophoretically distinct isoenzymes, with pI values of 6.3, 6.58, 6.63 and 6.88. The native enzyme has been crystallised in a form suitable for high resolution X-ray crystallographic studies. The crystals diffract to better than 0.19 nm spacing and are relatively stable to irradiation with X-rays. The space group is P6(1)22 (or P6(5)22) with cell dimensions a = b = 10.21 nm, c = 29.32 nm and the crystal probably contains a single monomer in the asymmetric unit. PMID- 3030754 TI - Overproduction and large-scale preparation of deoxyuridine triphosphate nucleotidohydrolase from Escherichia coli. AB - A recombinant plasmid, pHW1, directing the overproduction of the enzyme deoxyuridine 5'-triphosphate nucleotidohydrolase (dUTPase, EC 3.6.1.23) from Escherichia coli has been constructed. A 1900-base DNA fragment carrying the structural gene for the enzyme (dut) has been recloned into a runaway replication vector that also carries the strong leftward promoter (pL) of bacteriophage lambda. Upon temperature shift, an E. coli strain carrying the new plasmid gives an increase in dUTPase activity of about 600-fold in rich medium compared to wild type bacteria. The 64-kDa protein corresponding to the mature form of the enzyme reaches 20% of the total protein content of the bacterial cell. Using this strain, a simplified procedure has been developed for the purification of dUTPase. The purification steps consist of extraction of the cytoplasmic proteins, ammonium sulfate precipitation, anion-exchange chromatography and gel filtration on FPLC. The new overproducing plasmid and the simplified purification procedure developed will make it possible to purify dUTPase in sufficient amounts for detailed characterization studies. PMID- 3030753 TI - The protein phosphatases of Drosophila melanogaster and their inhibitors. AB - Protein phosphatases-1, 2A and 2B have been identified in membrane and soluble fractions of Drosophila melanogaster heads. Similarities between Drosophila and mammalian protein phosphatase-1 included specificity for the beta subunit of phosphorylase kinase, sensitivity to inhibitor-1 and inhibitor-2, inhibition by protamine, retention by heparin-Sepharose and selective interaction with membranes. In addition, an inactive form of protein phosphatase-1, termed protein phosphatase-1I, was detected in the soluble fraction that could be activated by preincubation with MgATP and mammalian glycogen synthase kinase-3. Inhibitor-2 partially purified from Drosophila had an identical molecular mass to its mammalian counterpart, and recombined with mammalian protein phosphatase-1 to form a hybrid protein phosphatase-1I. Similarities between Drosophila and mammalian protein phosphatase-2A included preferential dephosphorylation of the alpha subunit of phosphorylase kinase, insensitivity to inhibitors-1 and -2, activation by protamine, exclusion from heparin-Sepharose and apparent molecular mass. A Ca2+-dependent calmodulin-stimulated protein phosphatase (protein phosphatase-2B) that was inhibited by trifluoperazine was identified in the soluble fraction. The remarkable similarities between Drosophila protein phosphatases and their mammalian counterparts are indicative of strict phylogenetic conservation and demonstrate that the procedures used to classify mammalian protein phosphatases have a wider application. Characterisation of the Drosophila phosphatases will facilitate genetic analysis of dephosphorylation systems and their possible roles in neuronal and behavioural plasticity in Drosophila. PMID- 3030755 TI - Induction, partial purification and characterization of a hamster fibroblast protein kinase activity that phosphorylates ribosomal protein S6. AB - When BHK cells were grown to confluence and the growth medium replenished, there was a large and rapid increase in the phosphorylation of ribosomal protein S6. In postribosomal extracts of these cells, prepared in the presence of glycerol 2 phosphate and EGTA, a ribosomal protein S6 kinase was detected. The increase in activity of this protein kinase broadly reflected the increase in phosphorylation of ribosomal protein S6 observed in vivo. This ribosomal protein S6 kinase activity was substantially purified by a combination of phosphocellulose, DEAE cellulose, Mono Q and heparin-Sepharose chromatography, and some of its characteristics were examined. When the products of phosphorylation of 40S ribosomal subunits by purified enzyme in vitro were analysed using two dimensional gel electrophoresis, monophosphorylated and diphosphorylated forms of ribosomal protein S6 were observed to be the predominant radioactively labelled species. PMID- 3030756 TI - Plasma-gelsolin-binding sites on the actin sequence. AB - Plasma gelsolin was cross-linked to fluorescently labeled actin in order to identify plasma-gelsolin-binding sites on the primary sequence of actin. Plasma gelsolin can be cross-linked to actin with 1-ethyl-3-[3-(dimethyl amino)propyl]carbodiimide (EDC), resulting in the formation of 1:1 and 1:2 plasma gelsolin:actin cross-linked complexes with apparent molecular masses of 130 kDa and 180 kDa respectively. The cross-linked complexes were isolated separately, partially digested with cyanogen bromide and subjected to sodium dodecyl sulfate gel electrophoresis to analyze fluorescent fragments. The electrophoretic pattern showed that fluorescent CNBr fragments obtained from a free actin molecule were all found in those obtained from both the complexes. Since the fluorescent probe was attached to an actin molecule through the penultimate cysteine residue (Cys 374), the agreement in the fluorescent patterns indicated that the NH2-terminal CNBr fragments of both the actin molecules were involved in cross-linking with plasma gelsolin. This was also suggested by hydroxylamine cleavage of the two complexes as the cleavage gave the fluorescent 41-kDa fragment which could not be produced unless plasma gelsolin was cross-linked at the NH2-terminal segment comprising 12 amino acids. Since EDC cross-links an amino group with a carboxyl group only when they are direct contact, the characteristic acidic amino acid residues at the NH2 terminus of actin are suggested to participate in binding plasma gelsolin. PMID- 3030758 TI - Pneumatosis intestinalis related to cytomegalo-virus infection--a new etiology? AB - Five patients who developed pneumatosis intestinalis in the course of a cytomegalo virus (CMV) infection after cadaveric kidney transplantation are described. Based on the fact that CMV is known to cause intestinal ulcers, we postulate a causal relationship between CMV and pneumatosis intestinalis. PMID- 3030757 TI - A new human species of aldolase A mRNA from fibroblasts. AB - A full-length cDNA aldolase A clone was isolated from a human fibroblast cDNA library and completely sequenced. Excluding the poly(A) tail, the clone covers 1095 base pairs (bp) of the coding region, plus 199 bp downstream for the termination codon and 146 bp upstream for the initiation codon, within a total of 1440 bp. Primer extension experiments performed with human cultured fibroblast mRNA indicate an elongated product of a further 40 bp. These results evaluated together with those obtained in a concurrent study concerning aldolase A mRNA isolated from human liver are direct evidence of aldolase A mRNA multiplicity in man. The data also suggest the existence in mammals of three different classes of aldolase A mRNA, which would account for tissue specificity and resurgence of foetal expression in tumors. PMID- 3030759 TI - CT evaluation of pancreatic cancer. Analysis of resected tumours. AB - Preoperative CT examination including dynamic scan was performed in 25 patients with pancreatic carcinoma. Dynamic scans, which clearly demonstrated the attenuation difference between the normal and neoplastic pancreatic tissue in 92% of cases, were superior to high dose contrast enhancement scans that failed to display the relatively hypodense component of tumours in 40%. Detailed comparison of CT interpretation and surgical or pathological findings revealed the usefulness of CT examination in the evaluation of tumour size and local invasion. However, low sensitivity in detection of metastatic lesions (range: 20-38%) indicates limitations of CT diagnosis. PMID- 3030760 TI - Perioperative blood transfusion and prognosis of resected stage Ia lung cancer. AB - Based on the experience of blood-related immunosuppression in kidney transplants, some retrospective studies have reported an adverse relationship between blood transfusion and survival after curative resection for cancer. In order to confirm these findings, we have retrospectively evaluated our population of resected stage Ia non-small cell lung cancers (years 1974-79). Two hundred and eighty three patients were included in this analysis: 65 underwent pneumonectomy (23%), 205 lobectomy (72%) and 13 sublobar resections (5%). Patients submitted to perioperative blood transfusions were 157 (55%), without major differences according to surgery or tumour extent. The cumulative survival at 8 yr was 40% for transfused patients and 41% for nontransfused, relapse-free survival was respectively 36% and 34%; no differences were detectable stratifying for the amount of blood transfused or the extent of operation. Our experience does not support the hypothesis of an adverse prognosis related to perioperative blood transfusion. PMID- 3030761 TI - Subrenal capsule assay of fresh human tumors: problems and pitfalls. AB - The 6 day subrenal capsule (SRC) assay was performed in normal mice using 20 fresh human non-small cell lung cancers and nine fresh ovarian cancers. Different multi-agent chemotherapy regimens administered intravenously on days 2 + 3 of the assay were evaluated for activity against these two tumor types. Macroscopic results measured via an ocular microscope showed high activity for some combinations as well as for single agents. However, when subjected to microscopic examination, the SRC implants in the untreated control animals did not show viable tumor growth on day 6. Detailed histologic evaluation of over 800 SRC grafts reveals an intense inflammatory and fibrotic reaction in the majority of the grafts. These results indicate that drug sensitivity patterns obtained with this assay using only macroscopic criteria do not correlate with actual tumor regression. Microscopic analysis of the grafts prior to transplant shows absence or only minimal presence of tumor in many cases which also contributes to the poor growth observed in these two tumors. PMID- 3030762 TI - The haemodynamic and humoral effects of treatment for one month with the angiotensin converting enzyme inhibitor perindopril in salt replete hypertensive patients. AB - We have studied the effects of treatment for one month with perindopril, 4 or 8 mg once daily, in seven hypertensive patients. Blood pressure was lowered from 164/93 mm Hg to 145/84 mm Hg by 4 mg of perindopril and after one month remained at 142/82 mm Hg. Neither postural hypotension nor tachycardia occurred. Inhibition of plasma angiotensin converting enzyme (ACE) lasting for over 24 h was achieved and there was a significant increase in plasma renin activity (PRA). Maximum plasma concentrations of the active metabolite of perindopril, S-9780, were detected four h after oral administration. After treatment for one month there was evidence of reduced sensitivity of plasma ACE to the action of the inhibitor. The plasma concentration of S-9780 required to produce 50% inhibition of plasma ACE rose from 2.4 ng X ml-1 following the first dose to 5.5 ng X ml-1 after one month. PMID- 3030763 TI - The effect of chronic captopril therapy on platelet angiotensin II receptor density and vascular responsiveness to angiotensin II infusion. AB - The density of angiotensin II (Ang II) receptors on the platelet and the vascular responsiveness to infused angiotensin II before and after two weeks of captopril therapy were examined in ten healthy male volunteers. There was a significant increase in blood flow to the forearm, but no significant changes in either the density of angiotensin II receptors or the pressor response to infused angiotensin II following captopril therapy. The study demonstrates that long term reduction of angiotensin II formation by captopril in man does not increase the responsiveness of the receptors to infused angiotensin, nor results in an "up regulation" of the angiotensin receptors. It also provides further evidence that some of the long term vasodilator effects of captopril may be mediated by mechanisms other than inhibition of angiotensin I (Ang I) converting enzyme. PMID- 3030765 TI - Effect on blood pressure and the renin-angiotensin system of repeated doses of the converting enzyme inhibitor CGS 14824A. AB - A new, orally active angiotensin converting enzyme (ACE) inhibitor, CGS 14824A, was evaluated in 12 healthy male volunteers. Two groups each of 6 volunteers were given 5 or 10 mg once daily p.o. for 8 days. Four hours after the first and the last morning doses, plasma angiotensin II, aldosterone and plasma converting enzyme activity had fallen, while blood angiotensin I and plasma renin activity had risen. Throughout the study, more than 90% inhibition of ACE was found immediately before giving either the 5 or 10 mg dose and 50% blockade was still present 72 h following the last dose. Based on the determination of ACE, there was no evidence of drug accumulation. No significant change in blood pressure or heart rate was observed during the course of the study. CGS 14824A was an effective, orally active, long-lasting and well tolerated converting enzyme inhibitor. PMID- 3030766 TI - Functional activity in vivo of effector T cell populations. III. Protection against Moloney murine sarcoma virus (M-MSV)-induced tumors in T cell deficient mice by the adoptive transfer of a M-MSV-specific cytolytic T lymphocyte clone. AB - The functional activity of Moloney murine sarcoma virus (M-MSV)-specific T lymphocytes in vivo was assayed by the i.v. injection of virus-specific T lymphocytes into T cell-deficient "B mice". Virus-specific T lymphocytes generated in mixed lymphocyte tumor cell cultures were transferred i.v. into syngeneic "B mice" injected simultaneously at a distant site with the virus. These experiments indicated that a low dose (1 X 10(6) cultured cells) of infused lymphocytes can afford protection. To define the T lymphocyte subpopulation which was active, Lyt-2+ lymphocytes were selected by "panning" on plastic petri dishes coated with anti-Lyt-2 monoclonal antibody, and Lyt-2- lymphocytes selected by treatment with anti-Lyt-2 monoclonal antibody and complement. The results indicated that a Lyt-2+ lymphocyte-enriched population was more efficient in conferring protection against M-MSV-induced tumors. To investigate if cytolytic T lymphocytes (CTL) alone had a protective effect, a M-MSV-specific CTL clone was transferred in the same model system. The results demonstrated that a M-MSV specific CTL clone prevented M-MSV-induced tumor growth and also induced the destruction of syngeneic Moloney murine leukemia virus (M-MuLV)-induced MBL-2 leukemic cells in the peritoneal cavity. However, the cell dose required to obtain protection using a CTL clone was higher than that which was effective when mixed lymphocyte tumor cell culture cells were used. To assess the ability of the transferred cells to home and to repopulate the lymphoid organs of the "B mice", the frequency of virus-specific CTL precursors in the spleen was evaluated by limiting dilution analysis. The results indicated that lymphocytes from mixed lymphocyte tumor cell cultures can be recovered from the spleens of "B mice" injected i.v. 25 days earlier. On the contrary, following the transfer of an active CTL clone, a very low frequency (less than 1/200,000 cells) of virus specific CTL precursors was present in the spleens of recipient animals. The same M-MSV-specific CTL clone did not yield protection against M-MSV-induced tumors or MBL-2 leukemic cells when injected i.v. into M-MuLV tolerant mice. PMID- 3030764 TI - Enhancement of the ACTH response to human CRH by pretreatment with the antiglucocorticoid RU-486. AB - The response of ACTH to an i.v. bolus injection of 100 micrograms human CRH at 09.00 h was investigated in five healthy men with and without pretreatment with the antiglucocorticoid RU-486 100 mg given orally 7 h before the injection of CRH. In all five subjects the plasma cortisol level immediately before CRH administration at 09.00 h was significantly higher after pretreatment with the antiglucocorticoid (17.1 vs 11.1 micrograms/100 ml). Despite this higher baseline cortisol level, in all subjects the maximal CRH-induced ACTH increase was more pronounced after RU-486 loading (delta max ACTH 39 vs 26 pg/ml). This observation proves that in man physiological concentrations of cortisol determine the response of the pituitary to CRH. Furthermore, the findings suggest reduced circulating glucocorticoid activity after administration of 100 mg RU-486, not completely compensated by an increase in plasma cortisol. PMID- 3030767 TI - Plasmodium falciparum-specific human T cell clones: evidence for helper and cytotoxic activities. AB - This report describes the isolation, and the phenotypic and functional characterization of Plasmodium falciparum-specific human T lymphocyte clones (TLC) obtained from 2 acutely infected and 4 clinically immune donors. Approximately one third of the TLC obtained from the acutely infected patients had the phenotype CD8+/CD4-. No such clones were obtained from the clinically immune donors. P. falciparum-specific, major histocompatibility complex restricted CD8+ clones can lyse unrelated tumor cell targets in the presence of anti-CD3 antibodies, suggesting that these clones have cytotoxic potential. Conversely no killing was obtained with two CD4+ P. falciparum-specific TLC, even in the presence of anti-CD3 antibody, In addition the helper function of a CD4+ clone was demonstrated in a T/B cell cooperation system. PMID- 3030768 TI - Effect of lipopolysaccharide on intracellular killing of Leishmania enriettii and correlation with macrophage oxidative metabolism. AB - The effect of lipopolysaccharide (LPS) on the lymphokine (LK)-dependent activation of murine peritoneal macrophages for intracellular killing of Leishmania enriettii parasites was investigated. Exposure to LPS alone did not induce macrophages to kill the parasite. In the presence of LK or recombinant interferon-gamma, however, which by themselves rendered the macrophages only weakly cytotoxic, considerable stimulation of intracellular parasite killing was achieved already at a LPS concentration of 1 ng/ml. The response to LPS was of the same magnitude in macrophages tested for intracellular killing as in parallel assays of extracellular cytolysis of target cells. Acquisition of leishmanicidal activity by macrophages exposed to LK and LPS correlated with stimulation of the respiratory burst, as shown by increased hexose monophosphate shunt levels, and priming for elevated chemiluminescence and O2- and H2O2 production. Polymyxin B blocked both this LPS-dependent metabolic activity and intracellular parasite destruction. Intracellular killing was, however, not solely dependent on oxidative metabolism of macrophages since in the absence of LK, LPS stimulated respiratory burst activity, yet no intracellular killing was observed, and triggering of the respiratory burst by phorbol myristate acetate or zymosan did not affect intracellular parasite survival. These results suggest that, in this experimental model, efficient intracellular parasite killing depends both on increased production of oxygen metabolites and on the availability of so far unidentified factor(s), the synthesis of which requires exposure of macrophages to both LK and LPS. PMID- 3030769 TI - Epstein-Barr virus mediates a switch in responsiveness to transforming growth factor, type beta, in cells of the B cell lineage. AB - The functional performance of the transforming growth factor beta (TGF beta), appears to depend on the target cell phenotype as well as in vitro culture conditions. We show here that Epstein-Barr virus (EBV)-infection may induce a change in responsiveness to TGF beta, since TGF beta inhibits traverse of the cell cycle of activated normal human B cells, but promotes cell proliferation of EBV-positive Burkitt's lymphoma cell lines as well as EBV-infected B cells. We present evidence that the switch in the responsiveness to TGF beta is mediated by EBV infection, irrespective of the proliferative status of target cells, and thus may contribute to the initiation as well as the maintenance of certain B cell neoplasias. PMID- 3030770 TI - Vasoactive intestinal polypeptide (VIP) potentiates the muscarinic stimulation of phosphoinositide turnover in rat cerebral cortex. PMID- 3030771 TI - Inhibition of gastric K+/H+-ATPase by acid-activated 2-((2 pyridylmethyl)sulphinyl)benzimidazole products. AB - The inhibitory effects of timoprazole- and omeprazole-derived metabolites were studied in different in vitro test systems in order to characterize the metabolites of substituted benzimidazoles originating from acid activation. Acidification of timoprazole and omeprazole to pH 1.0 markedly increased the inhibitory potency on gastric K+/H+-ATPase. The timoprazole-derived tetracyclic thiol and radical were found to be equally or more potent on the K+/H+-ATPase than the mother compounds dissolved at pH 1.0. Kinetic studies with omeprazole sulphide revealed a competitive inhibition of the K+/H+-ATPase with respect to K+. The mercaptan dithiothreitol reversed the inhibitory effect of omeprazole, acidified timoprazole and the timoprazole-derived radical in the parietal cell and K+/H+-ATPase preparation. In contrast, the inhibitory effect of omeprazole sulphide and the timoprazole-derived thiol could not be reversed by dithiothreitol. Wash-out experiments indicated that acidified timoprazole and the tetracyclic compounds interact irreversibly with the K+/H+-ATPase, which contrasts with the properties of timoprazole in the parietal cell preparation. It is concluded from these data that neither the tetracyclic compounds nor the sulphide act as the 'active principle' of substituted benzimidazoles in the parietal cell preparion. PMID- 3030772 TI - Endothelium-dependent increases in rat gastric mucosal hemodynamics induced by acetylcholine and vagal stimulation. AB - The role of vascular endothelial cells in the vagal control of hemodynamics was studied in rat gastric mucosa. Vagal stimulation and intra-arterial administration of acetylcholine and of papaverine increased hemoglobin (Hb) and oxygen saturation of hemoglobin (SO2) in the gastric mucosa. The increases induced by vagal stimulation were reduced but not abolished by atropine. The responses to acetylcholine and vagal stimulation were reduced by quinacrine, p bromophenacyl bromide and nordihydroguaiaretic acid, while indomethacin had no effect. Intra-arterial infusion of collagenase removed the endothelial cells from submucosal vasculatures and depressed the increase in mucosal hemodynamics in response to acetylcholine and vagal stimulation. The response to papaverine was not depressed in rats treated with quinacrine or collagenase. These results suggest that the increase in gastric mucosal blood flow induced by acetylcholine or vagal stimulation is mediated by the endothelium-derived relaxing factor. PMID- 3030773 TI - Mechanism of antagonism of thromboxane receptors in vascular smooth muscle. AB - The mechanism and profile of antagonism of thromboxane receptors were studied in isolated perfused cat coronary arteries and in rat aortic rings. In cat coronary arteries, the thromboxane receptor antagonist (TxRA) BM-13,505 at concentrations from 3 to 300 ng/ml, significantly attenuated the vasoconstrictor effects of both a thromboxane A2 analog (CTA2) and an endoperoxide analog (U-46,619) and did not alter the constrictor responses to leukotriene D4, arginine vasopressin, or angiotensin II. In rat aortic rings precontracted by CTA2 or U-46,619, the effective threshold concentration of BM-13,505 for relaxation was 5 ng/ml. Lower concentrations of BM-13,505 exerted no relaxation, and higher concentrations exhibited faster relaxation to the precontracted baseline levels. This relaxation was not observed in aortic rings precontracted by norepinephrine or angiotensin II. The action of TxRA was not influenced by the absence of the endothelium or by pretreatment with a selective guanylate cyclase inhibitor, methylene blue. Also, thromboxane receptor antagonists do not appear to block thromboxane induced constriction by action as free radical scavengers. It can be concluded that replacing thromboxane A2 on the vascular receptor by a TxRA is the main factor responsible for the antagonism of thromboxane induced vasoconstriction in vascular smooth muscle preparations, not, the presence of the endothelium, activation of guanylate cyclase, or scavenging of superoxide free radicals. PMID- 3030774 TI - Pre- and postjunctional alpha-adrenoceptor-mediated effects of prazosin, methoxamine and 6-fluoronoradrenaline in blood-perfused canine skeletal muscle in situ. AB - Pre- and postjunctional effects of the alpha 1-selective adrenoceptor antagonist prazosin and the alpha 1- and alpha 2-selective adrenoceptor agonists methoxamine and 6-fluoronoradrenaline, respectively, were studied in skeletal muscle in situ. Prazosin reduced the vasoconstriction and enhanced the overflow of endogenous noradrenaline elicited by sympathetic nerve stimulation (1-4 Hz, 2 min); the threshold concentration was 10-100 times lower for postjunctional than for prejunctional alpha-adrenoceptor blockade. The enhancement of noradrenaline overflow by prazosin was not inversely frequency-dependent, as shown elsewhere for alpha 2-adrenoceptor antagonists. Thus, different mechanisms may be involved. Inhibition of prostaglandin synthesis by diclofenac did not alter the stimulation evoked noradrenaline overflow, indicating a minor importance of prostaglandin mediated transjunctional mechanisms in the modulation of noradrenaline overflow. Methoxamine and 6-fluoronoradrenaline elevated the basal vascular tone and, at higher concentrations, reduced the stimulation-evoked noradrenaline overflow. Methoxamine was 20 times more selective than 6-fluoronoradrenaline for postjunctional receptors. Our results are compatible with a pre- and postjunctional localization of alpha 2-adrenoceptors and a predominantly, but not exclusively, postjunctional localization of alpha 1-adrenoceptors. The postjunctional selectivity for prazosin was less marked than previously reported from in vitro studies. Hence, care should be taken when extrapolating in vitro findings to the more complex in vivo situation. PMID- 3030775 TI - Gonadotropin- and estrogen-induced increase of peripheral-type benzodiazepine binding sites in the hypophyseal-genital axis of rats. AB - Peripheral-type benzodiazepine binding sites (PBS) were demonstrated in the cell membranes of various organs (ovary, uterus, oviduct, pituitary and kidney) of mature and immature female rats by using the PBS-specific ligand [3H]PK 11195. The equilibrium dissociation constants of [3H]PK 11195 for PBS in mature rats ranged from 3 to 4 nM. The specific binding of [3H]PK 11195 (2 nM) in the hypophyseal-genital axis of immature (19-27 days old) female rats was found to be significantly increased in the ovary and uterus, concurrently with the increase in age. Administration of pregnant mare serum gonadotropin or diethylstilbestrol to immature rats increased the density of PBS in the ovary and uterus 2- to 3 fold but no change was found in the kidney. The affinity of [3H]PK 11195 to these tissues did not change following hormonal treatment. These results suggest that gonadotropin and estrogen are involved in the induction of PBS in the organs of the hypophyseal-genital axis in female rats. PMID- 3030776 TI - Evaluation of the supraspinal analgesic activity and abuse liability of ethylketocyclazocine. AB - Rats were trained to escape from aversive electrical brain stimulation delivered to the midbrain reticular formation (MRF) or to obtain rewarding intracranial stimulation delivered to the medial forebrain bundle (MFB). The threshold for escape behavior was determined by a modification of the psychophysical method of limits. Likewise, reward thresholds were determined by a rate-free psychophysical method. Acute administration of ethylketocyclazocine (EKC) (0.06-1.0 mg/kg s.c.) raised escape threshold in a dose-dependent manner while having no effect on non specific measures of sedation or motor impairment. This suggests that opioids with kappa receptor activity specifically modulate pain at a supraspinal level. Administration of EKC had no effect on the threshold for rewarding intracranial stimulation to the MFB suggesting that EKC has low potential for abuse. PMID- 3030777 TI - Chronic treatment with lorazepam and FG 7142 may change the effects of benzodiazepine receptor agonists, antagonists and inverse agonists by different mechanisms. AB - Treatment of mice with lorazepam 10 mg/kg p.o. or FG 7142 40 mg/kg i.p. once a day for 14 days changed the effects of benzodiazepine (BZ) receptor ligands injected acutely on the threshold of pentylenetetrazol (PTZ)-induced seizures. The effects of the two pretreatments differed qualitatively as well as quantitatively. Lorazepam elicited a shift in the effects of all BZ receptor ligands tested, whereby the agonists lorazepam and ZK 93423 now acted like partial agonists given acutely, the partial agonist ZK 91296 acted like an antagonist and the antagonists Ro 15-1788 and ZK 93426 like partial inverse agonists. The proconvulsant effects of the partial inverse agonist FG 7142 and the full inverse agonist DMCM on the PTZ-induced seizures did not change. However, FG 7142 became a full inverse agonist i.e. became convulsant, and DMCM may have increased in potency as a convulsant. After FG 7142 pretreatment lorazepam and ZK 93423 behaved like partial agonists given acutely whereas there was no change in effect for ZK 91296, Ro 15-1788 and ZK 93426. FG 7142 became convulsant (i.e. kindling occurred) and the potency of DMCM as a convulsant was non-significantly increased, while their proconvulsant effects with respect to PTZ-induced seizures were not altered. The fact that the effects of the two very different pretreatments on the BZ receptor ligand continuum were in the same direction may be explainable by assuming two different mechanisms, both of which may involve the GABA receptors. PMID- 3030778 TI - Autoradiographic localization of [3H] [MePhe3,D-Pro4]morphiceptin ([3H]PL017) to mu opioid receptors in rat brain. PMID- 3030779 TI - Clonidine binds to imidazole binding sites as well as alpha 2-adrenoceptors in the ventrolateral medulla. AB - Binding sites labeled by [3H]p-aminoclonidine ([3H]PAC) were characterized in bovine brain membranes prepared from the ventrolateral medulla, the probable site of the antihypertensive action of clonidine and analogs. Comparison was made with [3H]PAC binding to membranes prepared from frontal cortex, which has been studied extensively. Saturation binding isotherms for [3H]PAC were similar in the two regions, although Bmax values were approximately two-fold lower in ventrolateral medulla relative to frontal cortex. Norepinephrine and other phenylethylamines displaced [3H]PAC from a maximum of 70% of the total sites in the ventrolateral medulla. The remaining 30% were norepinephrine-insensitive, non-adrenoceptor sites which displayed high affinity for imidazole compounds. Ligand selectivity differed markedly between ventrolateral medulla and frontal cortex, since some imidazole compounds which potently inhibited [3H]PAC binding in the ventrolateral medulla had no effect in frontal cortex. Imidazole binding sites may mediate, in part, the hypotensive action of clonidine and other imidazole compounds in the ventrolateral medulla. These sites may also participate in the functions of a putative endogenous clonidine-like substance. PMID- 3030780 TI - Autoradiographic analysis of receptors on vascular endothelium. AB - Receptor autoradiography was used to examine the distribution of muscarinic cholinoceptors ([3H]QNB), alpha 2-adrenoceptors ([3H]rauwolscine), beta adrenoceptors ([125I]CYP) and substance P receptors ([125I]BHSP) in rabbit aorta, pulmonary artery, rat aorta, dog aorta, splenic, renal and coronary arteries, bovine aorta and coronary arteries. Muscarinic cholinoceptors and alpha 2 adrenoceptors were not associated with endothelium in any of the blood vessels examined. Substance P receptors were found on endothelium in dog renal but not bovine coronary arteries, and beta-adrenoceptors were found on endothelium in dog coronary arteries but not bovine aorta. The results suggest that endothelium dependent relaxation can result either from activation of receptors located directly on the endothelial cells or, as is the case for ACh, by an indirect mechanism via activation of receptors located on the vascular smooth muscle. PMID- 3030781 TI - Opposite effects of mu and kappa opiates on the firing-rate of dopamine cells in the substantia nigra of the rat. AB - Single unit extracellular recording was carried out in rats to compare the actions of a mu receptor agonist (morphine) to a kappa receptor agonist (U50,488) in the substantia nigra pars compacta (SNC). Intravenously administered morphine and U50,488 exhibited opposite effects: morphine excited SNC dopamine cells, and U50,488 inhibited them. Both effects were blocked by naloxone, although significantly more naloxone was necessary to block the effect of the kappa agonist. Local administration of U50,488 in the caudate also inhibited the firing of SNC dopamine cells. These results are discussed in terms of the natural functions of the prodynorphinergic straiatonigral projection. PMID- 3030782 TI - Two independent effects of cortisol on chicken liver. AB - Activities of two key enzymes of glycogen metabolism have been measured after an acute administration of cortisol to 3d-old chickens. Glycogen synthase activity is enhanced 2-3 hours after a cortisol injection, and this activation is blocked by use of protein synthesis inhibitors. Glycogen phosphorylase activity is enhanced at an early stage, and this effect is not suppressed by protein synthesis inhibitors. Liver cAMP levels are not increased concomitantly with this early activation of glycogen phosphorylase; indeed they are depleted. These results point to the existence of an effect of cortisol previous to and independent of its nuclear interaction, and not mediated by an activation of the membrane adenylate cyclase. PMID- 3030783 TI - Rat hypothalamus contains an antimitogenic factor acting via benzodiazepine receptors. AB - The effect of crude rat hypothalamic extracts on the lymphocyte proliferation in vitro was investigated. It was found that hypothalamic extracts (HE) significantly inhibited the 3H-thymidine incorporation by the mouse spleen lymphocytes cultured in vitro. The factor (or factors) responsible for the antimitogenic effect is thermostabile and partially destroyable by trypsin digestion. The latter observation suggests that it is probably a peptide. Diazepam had been found to exert a similar antimitogenic effect on mouse spleen lymphocytes as HE. The putative endogenous benzodiazepine (BZD) receptor ligand is also assumed to be a peptide. The working hypothesis that the antimitogenic effect of HE depends on endogenous substance acting via BZD receptor was proposed. To test such a possibility, the interaction of HE and specific BZD receptor ligands Ro-15-1788 and Ro-5-4864 was investigated. It was found that Ro 5-4864 alone suppressed the lymphocyte proliferation, and the joint antiproliferative effect of HE and Ro-5-4864 was not additive. Ro-15-1788 alone did not influence the lymphocyte proliferation but abolished the antiproliferative effect of HE. These data suggest that antimitogenic factor present in HE acts via BZD receptors. PMID- 3030785 TI - Effects of stimulated adrenocortical activity on the concentration of thyroxine and triiodothyronine in blood serum of piglets. AB - The effects of elevated adrenocortical activity, induced by cold exposure, weaning or starvation, or exogenous adrenocorticotrophin (ACTH) on the concentrations of thyroxine (T4) and triiodothyronine (T3) in blood serum were investigated in 99 sucking piglets and weaners, divided into five experiments. No changes in concentrations of circulating iodothyronines were recorded during a 3 hour exposure of 3- to 4-day-old piglets to ambient temperature of 8-12 degrees C. Two days after weaning of 4-week-old piglets the concentration of T4 increased. At the same time a decrease of the concentration of T3 was observed in their starving littermates. Two hours after the administration of ACTH, the concentration of both iodothyronines decreased, that of T4 nonsignificantly. 17 hours after the administration of the second of two doses of ACTH, i.e. at the time when the concentration of corticosteroids approached initial values, the blood concentration of iodothyronines both in sucking piglets and in weaners was elevated, that of T4 nonsignificantly. We suggest that for the changes of T4 and T3 concentrations in blood sera of stressed animals both specific stressor effects and circulating corticosteroids are responsible. The suppressive action of corticosteroids seems to be limited to the period of adrenocortical stimulation only. PMID- 3030784 TI - The biochemical mechanisms of the stimulating action of estradiol on the growth of mammary tumours. AB - The mechanisms of stimulating effect of estradiol on the growth of hormone dependent mammary tumours in experimental animals were studied. Estradiol receptors were detected in plasmatic membranes of estradiol-dependent tumour cells. Estradiol-independence of tumours was found to be a result of the impairment of the estradiol receptors in the plasmatic membranes, intracellular receptors, or of their ability to interact with the chromatin of the target cells. It was shown that the interaction of receptors of plasmatic membranes of hormone-dependent tumour cells with estradiol results in activation of the membrane adenylate cyclase and in a short-term rise of a cAMP content. A treatment of the hormone-dependent tumours with ovarian hormones brings about an increase in the activity of the cAMP-dependent protein kinases in cell nuclei, which points to their translocation from cytosol. Under effect of estradiol the phosphorylation of nuclear proteins at the serine, threonine and tyrosine residues increases. PMID- 3030786 TI - Superior efficacy of pulsatile versus continuous administration of ovine corticotropin releasing hormone in healthy man. AB - The effect of continuous (50 micrograms and 100 micrograms/4 hrs) versus pulsatile (6.25 micrograms every 30 minutes equivalent to a total dose of 50 micrograms during 4 hours) administration of ovine corticotropin releasing factor (oCRF) was investigated in 6 healthy, male subjects. A rise in the plasma concentrations of ACTH and of cortisol was observed during the intermittent, but not during the continuous infusion of 50 micrograms oCRF/4 hours. Comparing the rise in the plasma concentrations of ACTH and cortisol above individual basal concentrations no differences were found between the continuous infusion of 100 micrograms oCRF and the pulsatile administration of 50 micrograms oCRF. Response to pulsatile (50 micrograms/4 hrs) and continuous (100 micrograms/4 hrs) administration of oCRF, but lack of response to continuous infusion at a rate of 50 micrograms/4 hrs was also reflected by the calculated mean sum of increments of cortisol above individual basal concentrations. These results indicate a superior efficacy of intermittent versus continuous administration of oCRF in healthy man. PMID- 3030787 TI - Association of three chicken proteins with the 34 kD target of Rous sarcoma virus tyrosine kinase. AB - Transformation of chicken embryo fibroblasts by avian retroviruses induces the tyrosine phosphorylation of a 34-39 kD cellular protein (p34). In vitro, p34 isolated from intestine interacts with F-actin in a Ca2+-dependent manner. We report here that, in the absence of Ca2+ chelators, three proteins co-purified with p34 extracted from a cytosolic or membrane fraction of chicken embryo fibroblasts; these two fractions account respectively for 10-20% and 50% of the total cellular p34. Isolated from the cytosoluble fraction of fibroblasts by sucrose gradient centrifugation and hydrophobic chromatography, p34 and the other proteins behaved as a homogeneous species upon non-denaturing gel electrophoresis, gel filtration, and CsCl density gradient centrifugation, thus indicating a strong association. Moreover, an analysis by electron microscopy following uranyl acetate staining revealed particles with a raspberry-like shape. This association was always disrupted by the calcium-chelating agent, EGTA. PMID- 3030788 TI - Transformation of DNA repair-deficient human diploid fibroblasts with a simian virus 40 plasmid. AB - Fibroblasts from patients with xeroderma pigmentosum (XP) complementation groups A, C, D, E, and G, as well as Bloom syndrome (BS) and Fanconi anemia (FA) have been transfected with a plasmid, pSV7, containing the early region of Simian virus 40 (SV40). All of the cultures exhibited cytologic changes characteristic of transformed cells and expressed T-antigen. They also contained integrated copies of DNA derived from the vector, and in several cases, extrachromosomally replicated DNA. Not all of the transfected cultures became immortalized. The transformed xeroderma pigmentosum (XP) cultures retained their UV-sensitive phenotype in all but one case. The BS and FA cell lines retained their characteristic phenotype. All of the cultures, except the BS cells, can be readily transfected with the plasmids, pSV2neo and pSV2gpt. PMID- 3030789 TI - Frozen raspberries and hepatitis A. AB - An outbreak of 24 cases of hepatitis A in Aberdeen was traced to a large hotel in the city by epidemiological investigation. Food-specific questioning of those affected, their fellow diners and hotel staff, coupled with serological studies, implicated raspberry mousse prepared from frozen raspberries as the source of the infection. The raspberries were probably contaminated at the time of picking. PMID- 3030791 TI - An adenosine diphosphate phosphohydrolase in limbal vasculature. PMID- 3030790 TI - Seroepidemiological studies on the occurrence of common respiratory infections in paediatric student nurses and medical technology students. AB - The occupational risk of acquiring minor respiratory infections for paediatric student nurses was estimated by performing serological examinations with influenza A, B, C, parainfluenza, mumps, respiratory syncytial virus, adenovirus and Mycoplasma pneumoniae at 6-month intervals over a period of 4 years in paediatric student nurses at two schools of nursing and students at one school of medical technology. Titre increases against all tested agents occurred 1.86 times more often in the student nurses than in the medical technology students, the most frequent agents in both groups being influenza A and B. No difference in the relative distribution of the agents could be verified in the two occupational groups. Data on the protective value of pre-infectious antibody levels for influenza A, B, and coronavirus OC43 and on the importance of the spread of single agents among classmates are presented. PMID- 3030792 TI - Evidence for gluconeogenesis in the amphibian retina. AB - Evidence for the existence of a gluconeogenic pathway was provided in the amphibian retina. It was found that [3H]glutamate was converted to [3H]glucose derived from [3H]glutamate was incorporated into glycogen. The rate for this incorporation was found to be essentially the same in both light- and dark adapted retinas: 0.147 vs. 0.142 nmol (mg protein X 2 hr)-1, respectively. However, the rate of incorporation was found to decline progressively with time. The rate for the incorporation of label derived from glutamate into glycogen was found to be considerably less than that for [3H]glucose: 10.2 nmol (mg protein X 2 hr)-1. The activity of a key gluconeogenic enzyme, fructose-1,6-bisphosphatase, also was demonstrated in retinal supernatants, approximately 1 nmol (mg X min)-1, and the activity of this enzyme was found to be inhibited both by adenosine monophosphate and by fructose-2,6-bisphosphate. PMID- 3030793 TI - Effect of age on parathyroid hormone and forskolin stimulated adenylate cyclase and protein kinase activity in the renal cortex. AB - The capacity of parathyroid hormone (PTH) to stimulate the renal production of 1,25-dihydroxyvitamin D declines with age. Since the action of PTH in the kidney is mediated by cAMP, we have examined the effect of PTH and forskolin on renal cortical adenylate cyclase in young (3 months), adult (13-15 months), and old (25 27 months) F344 rats. PTH-dependent adenylate cyclase, measured as cAMP accumulation in cortical slices, was reduced in adult and old rats compared to young rats over a PTH concentration range of 0.015-15 units/ml. There was no difference in PTH-dependent adenylate cyclase activity between adult and old rats. Renal plasma membrane preparations demonstrated similar changes in PTH stimulated adenylate cyclase activity. There was no difference in calcitonin, forskolin, or guanyl-5-ylimidodiphosphate (Gpp(NH)p) stimulation of adenylate cyclase in plasma membranes from each age group, suggesting that the defect lies in the membrane receptor for PTH. The decreased adult sensitivity to PTH could be reversed by thyroparathyroidectomy. PTH stimulation of cytosolic protein kinase activity did not change with age. These results suggest that the decrease in PTH dependent adenylate cyclase is due to the alterations at the level of PTH receptor. These alterations may be in response to the increase in serum PTH seen in these animals with increasing age. PMID- 3030794 TI - Reflex pathways from group II muscle afferents. 2. Functional characteristics of reflex pathways to alpha-motoneurones. AB - The convergence of group II muscle afferents on interneurones in reflex pathways has been elucidated by investigating interaction in transmission to motoneurones. Recording was also made from interneurones activated from group II afferents. Maximal group II EPSPs evoked in motoneurones from different muscles (extensors or flexors and extensors) did not summate linearly but with a deficit of 35-40%. The corresponding deficit in summation with Ia EPSPs was 7%. It is suggested that the difference in deficit is caused largely by occlusion due to shared interneuronal discharge zones and that it gives an approximate minimal measure of the convergence of group II afferents from different muscles on the interneurones. Tests with weak group II volleys from different muscles gave no or little evidence for spatial facilitation in the disynaptic excitatory pathway to flexor motoneurones, and there was no or little temporal facilitation of transmission in this pathway. It is suggested that group II excitation of the interneurones in this pathway depends on few afferents giving large unitary EPSPs. Convergence of cutaneous afferents and joint afferents on the interneurones was evidenced by spatial facilitation from these afferents of group II transmission to motoneurones. Convergence on interneurones in the trisynaptic inhibitory pathway from group II afferents to extensor motoneurones was also investigated with the spatial facilitation technique. There was convergence on common interneurones of group II afferents from different muscles (extensors or flexors and extensors) and from cutaneous afferents as well as joint afferents. Trisynaptic group II IPSPs, including those depending on spatial facilitation from different muscles were resistant to recurrent depression from motor axon collaterals and are therefore not mediated by the reciprocal Ia inhibitory pathway. Interneurones with monosynaptic group II EPSPs were recorded from in the dorsal horn and intermediate region. Graded stimulation revealed large unitary EPSPs from few group II afferents. The EPSP evoked by a single group II afferent may produce firing (extracellular recording). Convergence of monosynaptic group II EPSPs from different muscles was rather limited but could be from flexors and extensors. Extensive multisensory convergence onto some of these interneurones was indicated by di- or polysynaptic EPSPs from group II and III muscle afferents, from joint afferents and from cutaneous afferents.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3030795 TI - Distribution of potentiation following short high frequency bursts to motoneurons of different rheobase. AB - Potentiation of composite EPSPs has been studied at group Ia fiber/alpha motoneuron connections using a short burst of conditioning stimuli [32 shocks with 6 ms interstimulus interval (ISI)]. Potentiation reached its peak (range 1.2 2.0 X control value) 100-150 ms following the burst. Potentiation decayed slowly and was still present 2 s after the burst. High frequency burst stimulation of a nerve to a synergist muscle did not potentiate the response to stimulation of the homonymous nerve. Three independent sets of measurements suggest that the time course of decline of potentiation depends on the amount of transmission during the potentiated state. First, connections on high rheobase motoneurons exhibited more peak potentiation than those on low rheobase motoneurons but potentiation decayed more rapidly in the former than in the latter. Second, increasing the frequency of the conditioning burst enhanced peak potentiation but the rate of decay of this potentiation also increased. Third, potentiated EPSPs exhibited more low frequency depression than unpotentiated ones at the same connection suggesting that low frequency stimulation during the potentiated state could elevate the rate of decay of potentiation. The short burst paradigm could cause peak potentiation similar in magnitude to that evoked by long, high frequency trains (studied here at the same connection) but with much less of an increase in latency and rise time of the potentiated EPSP. The magnitude of potentiation was unrelated to changes in EPSP latency and rise time. These findings indicate that potentiation can act to modulate EPSP amplitude under conditions of normal motor behaviour when spindle afferents fire in patterns similar in duration and frequency to those used in the present experiments. PMID- 3030796 TI - Nitroglycerin-induced desensitization of vascular smooth muscle may be mediated through cyclic GMP-disinhibition of phosphatidylinositol hydrolysis. AB - The purpose of this study was to investigate the hypothesis that nitroglycerin induced desensitization of vascular smooth muscle is mediated through cyclic GMP disinhibition of phosphatidylinositol hydrolysis. Norepinephrine-induced contraction and increased levels of inositol monophosphate, a measure of phosphatidylinositol hydrolysis, in rat aorta. Prior treatment with nitroglycerin inhibited both the norepinephrine-induced contraction and the elevated levels of inositol monophosphate to the same relative magnitude. The nitroglycerin-induced inhibition of contraction and inositol monophosphate formation were prevented in tissues desensitized with nitroglycerin. These results suggest that: nitroglycerin may inhibit vascular smooth muscle contraction through cyclic GMP inhibition of phosphatidylinositol hydrolysis and desensitization to the relaxant effects of nitroglycerin may be due to disinhibition of the hydrolysis. PMID- 3030797 TI - Enzyme markers of collagen synthesis in carbon tetrachloride-induced fibrosis and during colchicine modification of CCl4-induced liver injury. AB - Serum galactosylhydroxylysyl glucosyltransferase (S-Glu-Gal-Hyl-Tase), liver galactosylhydroxylysyl glucosyltransferase (L-Glu-Gal-Hyl-Tase), liver hydroxylysyl galactosyltransferase (L-Gal-Hyl-Tase), and liver prolyl hydroxylase (L-PH) activities were measured in rats during the development of CCl4-induced cirrhosis (0.2 ml of 33% CCl4 in light mineral oil two times weekly for 10 weeks followed by 6 weeks of no treatment). Serum and liver markers of collagen synthesis increased in a time-dependent manner reaching maximum activity at 6 weeks (S-Glu-Gal-Hyl-Tase, two times; L-PH, two times). These enzyme levels returned to normal during the 4-week recovery period. In a separate 4-week experiment, colchicine (10 micrograms/rat/day) was administered with CCl4. Colchicine prevented the increase in S-Glu-Gal-Hyl-Tase, L-Glu-Gal-Hyl-Tase, and L-Gal-Hyl-Tase induced by CCl4 and resulted in a smaller increase in L-PH. These results demonstrate that S-Glu-Gal-Hyl-Tase elevation occurs following CCl4 because of increased liver collagen synthetic activity and the hepatocellular injury produced by CCl4. PMID- 3030799 TI - Biochemical effects of gentamicin on rat kidney cortex. I. Analytical subfractionation of control tissue. AB - As a first step in studies of the molecular mechanism(s) underlying gentamicin toxicity, rat kidney cortex has been subfractionated using differential centrifugation. An analytical, rather than preparative approach was used. DNA was used as a marker for the nuclei, cytochrome oxidase for mitochondria, acid phosphatase for lysosomes, catalase for peroxisomes, NADPH-cytochrome c reductase for the endoplasmic reticulum, p-nitrophenyl-alpha-mannosidase (at pH 5.5) for the Golgi apparatus, AMPase for the plasma membrane in general, and alkaline phosphatase for the brush border, and lactate dehydrogenase for the cytosol. In addition, electron microscopy was performed on the subfractions obtained. The distributions of subcellular markers obtained here for the rat kidney cortex closely resemble the corresponding distributions reported for rat liver. This procedure can now be used to look for biochemical and/or toxic changes which might be reflected in an altered distribution pattern for marker enzymes. PMID- 3030798 TI - Changes in collagen metabolism and proteinolysis after repeated inhalation exposure to ozone. AB - To study the changes in collagen metabolism that occur in the pathogenesis of pulmonary fibrosis, female rats were exposed to 0, 0.57, and 1.1 ppm ozone for 19 hr/day for 11 days and sacrificed 12 or 60 days after initiation of exposure. The lungs of rats sacrificed at 12 days after initiation of exposure to 1.1 ppm had interstitial pneumonia characterized by a mixed inflammatory cell infiltrate, type II cell hyperplasia, and fibroplasia, a proliferation of the collagen producing cells; increased cathepsin D and macrophage elastase activity, indicating macrophage-induced proteinolysis; a reduced percentage of the increased collagen production that was ultrafilterable, indicating a decreased rate of intracellular degradation of newly produced collagen prior to its secretion; and increased lavage fluid hydroxyproline, indicating turnover of extracellular collagenous matrix. Reduced intracellular collagen degradation correlated directly with both increased net collagen production and fibroplasia in rats exposed to 1.1 ppm ozone for 11 days. These changes preceded an increased total lung collagen and the development of modest fibroplasia and fibrosis in the alveolar duct regions by 60 days after the 1.1 ppm ozone exposure was initiated. PMID- 3030800 TI - Biochemical effects of gentamicin on rat kidney cortex. II. Analytical subfractionation after short-term, high-dose treatment. AB - As a first step in studies on the molecular mechanism(s) underlying gentamicin toxicity, the effect of treating rats with this aminoglycoside antibiotic (100 mg/kg once or twice daily for 3 days) on the analytical subfractionation of the kidney cortex has been examined. DNA was used as a marker for the nuclei, cytochrome oxidase for mitochondria, acid phosphatase for lysosomes, catalase for peroxisomes (with reservations; see the companion paper), NADPH-cytochrome c reductase for the endoplasmic reticulum, p-nitrophenyl-alpha-mannosidase (at pH 5.5) for the Golgi apparatus, AMPase for the plasma membrane in general and alkaline phosphatase for the brush border, and lactate dehydrogenase for the cytosol. In addition, the presumptive lysosomal hydrolases N-acetyl-beta-D glucosaminidase, p-nitrophenyl-alpha-mannosidase (at pH 4.5), cathepsin D, and DNase II were monitored. Electron microscopy was also performed on the subfractions obtained. The only significant biochemical changes brought about by gentamicin treatment were that N-acetyl-beta-D-glucosaminidase demonstrated both a greater total activity and a larger enrichment in the 104,000gav pellet, while p-nitrophenyl-alpha-mannosidase at pH 4.5 demonstrated the same total activity and a greater enrichment in the 104,000gav pellet. Since myeloid bodies were shown by electron microscopy to sediment primarily with the 500gav and 10,000gav pellets, the biochemical changes seen cannot be associated with these morphological structures. These findings suggest that selective changes in a certain subpopulation(s) of lysosomes or in certain lysosomal enzymes may be involved in the early stages of gentamicin toxicity. On the other hand, no lysosomal membrane damage was observed here, since both the latency of acid phosphatase and the recovery of this activity in the soluble cytosol were unchanged. The present investigation may also have relevance for the dosage and duration of gentamicin treatment chosen in clinical situations. PMID- 3030802 TI - [Comparative study of the analgesic, sedative and myorelaxant effects of GABA positive preparations]. AB - It was shown in experiments on rats that the analgesic effect of GABA-positive drugs is followed by sedative and myorelaxant manifestations, however there is no direct relationship between the effects. Bicuculline significantly attenuates analgesia induced by THIP, gamma-acetylenic-GABA and depakin in the test of vocalization but enhances their analgesic activity in the tail-flick test. The involvement of individual subtypes of GABA receptors in mediation of analgesic and general depressant effects of GABA-positive drugs was discussed. PMID- 3030801 TI - Manuals of food quality control. 7. Food analysis: general techniques, additives, contaminants, and composition. PMID- 3030803 TI - A simple, one-step purification of cytochrome b from the bc1 complexes of bacteria. PMID- 3030804 TI - Glucose regulates preproinsulin messenger RNA levels in a clonal cell line of simian virus 40-transformed B cells. AB - In HIT-T15 cells grown in the absence of glucose, Northern blot analysis of total RNA revealed a major 0.5 kb preproinsulin (ppI) mRNA transcript which co-migrated with the mature transcript from a human insulinoma. In 4 h tissue cultures, glucose (2-20 mM) stimulated HIT cell ppI mRNA levels in a markedly dose dependent manner. Glucose-stimulated ppI mRNA was (i) inhibited by actinomycin D, suggesting that regulation may be in part transcriptional, and (ii) potentiated by agents known to activate B cell protein kinases. HIT cells represent a unique model for investigating long term regulation of insulin gene expression and biosynthesis. PMID- 3030805 TI - Biologically significant scavenging of the myeloperoxidase-derived oxidant hypochlorous acid by ascorbic acid. Implications for antioxidant protection in the inflamed rheumatoid joint. AB - Ascorbic acid, at physiological concentrations, can scavenge the myeloperoxidase derived oxidant hypochlorous acid at rates sufficient to protect alpha 1 antiprotease against inactivation by this molecule. The rapid depletion of ascorbic acid at sites of inflammation, as in the inflamed rheumatoid joint, may therefore facilitate proteolytic damage. PMID- 3030806 TI - Identification of a new 84/82 kDa calmodulin-binding protein, which also interacts with actin filaments, tubulin and spectrin, as synapsin I. AB - A new 84/82 kDa calmodulin-binding protein, which also interacts with actin filaments, tubulin and spectrin, was purified from the bovine synaptosomal membrane. The binding of calmodulin to this protein was Ca2+-dependent, and was inhibited by trifluoperazine, the association constant being calculated to be 2.2 X 10(6) M-1. Maximally, 1 mol of calmodulin bound to 1 mol of the purified protein. This protein was phosphorylated by both kinase II (Ca2+- and calmodulin dependent kinase) and cyclic AMP-dependent kinase. In addition, antibody against this protein was demonstrated to have an immunological crossreactivity with synapsin I in the synaptosomal membrane. PMID- 3030807 TI - Bimodal regulation of secretion by cytoplasmic Ca2+ as demonstrated by the parathyroid. AB - Bovine parathyroid cells were used to study parathyroid hormone (PTH) release and the cytoplasmic Ca2+ concentration (Cai2+). When the extracellular Ca2+ concentration was decreased from 3.0 to 0.5 mM, perifused cells reacted with rapid stimulation of PTH release. However, a further reduction of extracellular Ca2+ to less than 10 nM resulted in prompt inhibition. Both effects were readily reversible. Using the intracellular Ca2+ indicator quin-2 also as a buffer for calcium it was possible to control Cai2+ within the 20-600 nM range. PTH release was found to increase with Cai2+ up to 200 nM but was gradually suppressed above this concentration. PMID- 3030808 TI - The human preproapolipoprotein C-II gene. Complete nucleic acid sequence and genomic organization. AB - The complete nucleic acid sequence of human preproapolipoprotein (apo) C-II has been determined from 2 apoC-II clones isolated from 2 different human genomic DNA libraries. The cloned fragments were approx. 14 and 18 kb long, and sequence analysis established that the apoC-II gene consists of 3338 nucleotides containing 3 intervening sequences of 2391, 167, and 298 bases. The first intron is located within the 5'-untranslated region of apoC-II and contains 4 Alu type sequences. The second intron interrupts the codon specifying amino acid - 11 of the apoC-II signal peptide. The last intron, which contains a 38 bp sequence which is repeated 6 times, interrupts the codon specifying for amino acid +44 of the mature apolipoprotein. PMID- 3030809 TI - Allopurinol and oxypurinol are hydroxyl radical scavengers. AB - Allopurinol is a scavenger of the highly reactive hydroxyl radical (k2 approx. 10(9) M-1 X s-1). One product of attack of hydroxyl radical upon allopurinol is oxypurinol, which is a major metabolite of allopurinol. Oxypurinol is a better hydroxyl radical scavenger than is allopurinol (k2 approx. 4 X 10(9) M-1 X s-1) and it also reacts with the myeloperoxidase-derived oxidant hypochlorous acid. Hence the protective actions of allopurinol against reperfusion damage after hypoxia need not be entirely due to xanthine oxidase inhibition. PMID- 3030810 TI - The family of human Na+,K+-ATPase genes. A partial nucleotide sequence related to the alpha-subunit. PMID- 3030811 TI - The measurement of free radical reactions in humans. Some thoughts for future experimentation. AB - The question as to whether free radical reactions are a major cause of tissue injury in human disease, or merely an accompaniment to such injury, is very difficult to answer because of lack of adequate experimental techniques. New techniques that are becoming available are discussed, with specific reference to their use in humans. PMID- 3030812 TI - Molecular cloning of Clostridium difficile toxin A gene fragment in lambda gt11. AB - Toxin A of Clostridium difficile has been purified and monospecific antiserum produced. A reliable procedure for isolation and restriction of C. difficile chromosomal DNA was developed which allowed for the construction of a genomic library in lambda gt11. Approx. 35,000 plaques were screened using anti-toxin A which resulted in the identification of one stable positive clone, lambda cd19. Verification of the immunological identity of the isolated toxin A gene fragment in lambda cd19 was determined by affinity purifying toxin A antibodies specific for lambda cd19 gene product, and using these selected antibodies to probe a Western blot of purified toxin A. The insert in lambda cd19 was demonstrated to be a 0.3 kb fragment by restriction digestion, and by hybridization of the clone to a chromosomal digest of C. difficile. The peptide coded for by the toxin A gene fragment in lambda cd19 was not cytotoxic for 3T3 mammalian tissue culture cells. PMID- 3030813 TI - Molecular cloning and characterization of a full-length cDNA clone for human plasminogen. AB - A human liver cDNA library enriched for full-length clones was screened for plasminogen cDNA using a synthetic 24-nucleotide probe derived from a reported partial cDNA sequence. 12 positive clones were identified and one of these was characterized in detail. The 2.7 kb insert contains the complete coding region. At 5 positions, it gives residues different from those reported in a previous amino acid sequence analysis of the protein. The present results show an extra Ile at position 65, Gln instead of Glu at positions 53 and 342, Asn at position 88 instead of Asp, and Asp at position 453 rather than Asn. In the 3'-non-coding region an extension of 29 bases is found which does not contain any structure compatible with a known polyadenylation signal. Instead, the consensus signal AATAAA is placed at a distance of 46 bases upstream of the poly(A)-tail. PMID- 3030814 TI - The three-dimensional similarity between a dimeric antiparallel extended structure and a beta-turn folded form of enkephalin. AB - The three-dimensional similarity between two fundamental conformations, a dimeric antiparallel extended structure and a type I' beta-turn folded form, of enkephalin was examined by computer graphic and empirical energy calculation methods. By the rotation of Tyr and Phe side chains, one half of the former structure could mimic the three-dimensional form of the latter without a large loss of conformational energy. This result provides a new idea for considering the conformation of enkephalin suitable for the multiple opioid receptors. The active conformation of enkephalin for mu- and delta-opioid receptors is discussed in the light of the present study. PMID- 3030815 TI - A comparison of the restrained molecular dynamics and distance geometry methods for determining three-dimensional structures of proteins on the basis of interproton distances. AB - A direct comparison of the metric matrix distance geometry and restrained molecular dynamics methods for determining three-dimensional structures of proteins on the basis of interproton distances is presented using crambin as a model system. It is shown that both methods reproduce the overall features of the secondary and tertiary structure (shape and polypeptide fold). The region of conformational space sampled by the converged structures generated by the two methods is similar in size, and in both cases the converged structures are distributed about mean structures which are closer to the X-ray structure than any of the individual structures. The restrained molecular dynamics structures are superior to those obtained from distance geometry as regards local backbone conformation, side chain positions and non-bonding energies. PMID- 3030816 TI - Cyclic AMP-like effects of polyamines on phosphatidylcholine synthesis and protein phosphorylation in human promyelocytic leukemia HL60 cells. Comparison with the effects of phorbol ester. AB - Spermine or putrescine increased cAMP levels through a catalase-sensitive mechanism, resulting in, most notably, a dephosphorylation of protein A (Mr 45,000, pI 5.15) and protein B (Mr 45,000, pI 4.9) and slightly increased phosphatidylcholine (PC) synthesis in HL60 cells. Exogenous dibutyryl cAMP mimicked the polyamine effects. 12-O-Tetradecanoyl phorbol-13-acetate (TPA) also promoted the protein dephosphorylation and PC synthesis, the effects augmented by R59022 and mimicked by exogenous 1-oleoyl-2-acetylglycerol. The effects of spermine (or dibutyryl cAMP) and TPA on PC synthesis were synergistic. It was suggested that cAMP-dependent protein kinase and protein kinase C might mediate, in an independent but inter-related manner, the effects of polyamines and TPA. PMID- 3030817 TI - Oxidation-reduction midpoint potentials of the molybdenum center in spinach NADH:nitrate reductase. AB - Oxidation-reduction midpoint potentials for the molybdenum center in assimilatory NADH:nitrate reductase isolated from spinach (Spinacia oleracea) have been determined at pH 7.0 in the presence of dye mediators using EPR spectroscopy to monitor formation of Mo(V). Values for the Mo(VI)/Mo(V) and Mo(V)/Mo(IV) couples were determined to be -8 and -42 mV, respectively. PMID- 3030819 TI - Two-electron reduction is required for rapid internal electron transfer in resting, pulsed and oxygenated cytochrome c oxidase. AB - The resting as well as the 420 nm and 428 nm forms of cytochrome oxidase have been studied in kinetic experiments with an excess of enzyme over reduced cytochrome c. No difference was found in the behavior of the two activated forms. With all three forms, a fraction of cytochrome a was reoxidized with a rate which was much lower than kcat. This suggests that intramolecular transfer to the dioxygen-reducing site occurs only if both cytochrome a and CuA are reduced. An initial rapid phase in the oxidation of cytochrome a in the pulsed and oxygenated enzymes is related to the presence of a three-electron-reduced dioxygen intermediate. The increased catalytic activity of pulsed and oxygenated oxidase can be explained on the basis of a shift in the redox equilibrium between cytochrome a and CuA. PMID- 3030818 TI - NMR study of the interaction between cytochrome b5 and cytochrome c. Observation of a ternary complex formed by the two proteins and [Cr(en)3]3+. AB - The interaction between horse cytochrome c and the tryptic fragment of bovine liver microsomal cytochrome b5 in the absence and presence of [Cr(ethylenediamine)3]Cl3 was studied by 1H NMR spectroscopy. The protein-protein interaction region on cytochrome b5 was found to be different from the [Cr(en)3]3+-binding region. The solvent-exposed propionate-bearing edge of the haem of cytochrome b5 is accessible to [Cr(en)3]3+ in the interprotein complex. PMID- 3030820 TI - Permeabilization of thymocytes by photon activation of erythrosin. AB - Thymocytes previously loaded with quin 2 were rapidly permeabilized by the photon activation of erythrosin and the rate of permeabilization monitored by measuring fluorimetrically the increasing saturation of quin 2 with calcium. The extent of permeabilization was assessed also by the loss of [3H]quin 2 from the thymocytes and penetration of the cells by eosin and trypan blue. Lactate dehydrogenase leakage from the permeabilized cells was markedly delayed compared to the rapid increase in permeability to calcium and quin 2. The rate of permeabilization was dependent upon the concentration of erythrosin, the duration of illumination, the presence of oxygen, and the temperature. These results are consistent with the rapid photochemical generation of highly reactive singlet oxygen which alters thymocyte membrane structure and permeability. PMID- 3030822 TI - The cytoskeletal protein vinculin is acylated by myristic acid. AB - In non-muscle cells the mechanism by which microfilament bundles interact with the plasma membrane is unclear. Vinculin, a 130 kDa protein found in adhesion plaques, has been postulated to have a role as a membrane anchor for microfilaments and we have investigated the biochemistry of this molecule in more detail. We report that a fraction of vinculin in chick embryo fibroblasts is acylated by myristic acid. This modification was present in both membrane-bound, cytoskeletal and cytosolic vinculin and thus did not determine preferential subcellular localisation. Myristic acid was also present in vinculin from cells transformed by Rous sarcoma virus. PMID- 3030821 TI - Adenosine and oxytocin reverse antagonism of cyclic AMP elevating agents to insulin activation of adipocyte pyruvate dehydrogenase. AB - ACTH, isoprenaline, forskolin, and dibutyryl cyclic AMP prevented insulin from stimulating adipocyte pyruvate dehydrogenase in the presence of adenosine deaminase. Antagonism was reversed by N6-phenylisopropyladenosine as well as oxytocin. The stimulatory effects of insulin, adenosine and oxytocin on adipocyte pyruvate dehydrogenase appear to be through (a) mechanism(s) which is (are) similar or related. PMID- 3030823 TI - In situ carcinomas of the breast. Types, growth pattern, diagnosis, and treatment. AB - The paper presents a prospective study comprising 40 consecutive patients with in situ carcinomas of the breast and two with atypical ductal hyperplasia (ADH) who underwent operation during a 2-year period at a single hospital. Out of the 40 in situ carcinomas 13 were of the lobular type (LCIS) and 27 of the ductal type (DCIS). They made up about 9% of all newly diagnosed breast cancers. Histologically a distinction could be made between three different growth patterns: microfocal, tumour-forming, and a diffuse form. With the exception of one case, the 26 microfocal growths (2 ADH, 13 LCIS, 11 DCIS) were accidental findings in otherwise benign breast biopsies, whereas the tumour-forming and diffuse forms (16 DCIS) were diagnosed in the great majority clinically and/or by mammography. Of the tumour-forming and diffuse DCIS 25% were demonstrated solely by mammography. The surgical treatment in the 26 patients showing microfocal changes was exclusively biopsy in 23, while three had mastectomy, because of a papillary focus in two and patient preference in one. Of six patients with tumour forming DCIS three had segmental resection and three mastectomy, the latter because of papillary foci, while all 10 with diffuse growth had mastectomy. On the basis of their experience of types and growth patterns, the authors set up a surgical strategy which might add new aspects to our knowledge about the biological nature of in situ lesions. PMID- 3030824 TI - Ductal carcinoma in situ (DCIS) of the breast--a therapeutic dilemma. AB - The incidence of ductal carcinoma in situ of the breast (DCIS) is low. Multifocality and difficulties in defining the extent of the disease are the main problems in relation to the therapy. Biologic characterization still is incomplete. Ablative treatment gives a 100% cure. Prospective trials are needed to evaluate breast conserving therapy. PMID- 3030825 TI - Estrogen carcinogenesis in Syrian hamster tissues: role of metabolism. AB - Evidence for a role of estrogen metabolism in hormonal carcinogenesis was obtained with the Syrian hamster as an in vivo model system. Both natural and synthetic estrogens are capable of inducing a high incidence of renal carcinomas in this species. A high incidence of hepatocellular carcinomas can also be induced in the hamster with synthetic estrogens such as ethinyl estradiol or diethylstilbestrol, provided alpha-naphthoflavone (ANF) is present in the diet. Although steroid receptor-mediated hormonal events appear to be intimately involved in the process of in vivo cell transformation of both tissues, certain observations strongly suggest that nonhormonal events are also important. Despite their potent estrogenic activity at the doses used, ethinyl estradiol and alpha zearalanol induce relatively low renal tumor incidences after 9.0 and 10.0 months of continuous treatment, respectively. A role for the metabolism of estrogens to reactive intermediates is also suggested by studies showing estrogen-induced renal tumorigenesis can be partially inhibited by concomitant administration of ANF or ascorbic acid. Consistent with this is the general correlation between the amount of catechol estrogen formed by a compound, as mediated by estrogen 2-/4 hydroxylase, and renal carcinogenicity data. Recently, additional supporting evidence has been obtained from studies involving the irreversible binding of reactive metabolites of steroidal or stilbene estrogens to hamster liver microsomal proteins. PMID- 3030826 TI - Photosynthesis of vitamin D in the skin: effect of environmental and life-style variables. AB - Exposure to sunlight continues to play a major role in providing adequate vitamin D nutrition for most of the population of the world, including those who live in countries that practice fortification of dairy, margarine, and cereal products with vitamin D. During exposure to sunlight, the high-energy UV photons (290-315 nm) penetrate the epidermis and photolyze 7-dehydrocholesterol (provitamin D3) to previtamin D3. Once formed, previtamin D3 undergoes a thermally induced isomerization to vitamin D3 that takes 2-3 days to reach completion. Melanin effectively competes with provitamin D3 for the UV radiation that enters the epidermis and limits its photolysis to previtamin D3. However, this is not the major factor that prevents excess production of vitamin D in the skin of people who are constantly exposed to sunlight. During the initial exposure to sunlight, provitamin D3 is efficiently converted to previtamin D3. However, because previtamin D3 is photolabile, continued exposure to sunlight causes the isomerization of previtamin D3, principally to lumisterol. Thus, no more than 10 20% of the initial provitamin D3 concentrations ultimately end up as previtamin D3. Aging, sunscreens, seasonal changes, time of day, and latitude also significantly affect the cutaneous production of this vitamin-hormone. PMID- 3030827 TI - Antigonadotropic effect of oxytocin on the isolated human corpus luteum. AB - From each of 43 women who had gynecologic laparotomy at different and well classified stages of the luteal phase of the cycle, the corpus luteum (CL) was excised, cut into pieces, and incubated for short time periods (30 to 240 minutes). Incubations were performed in the absence and presence of human chorionic gonadotropin (hCG) and different concentrations of oxytocin ([OT], 0.1 to 1000 mIU/ml). After incubation, the tissue levels of cyclic adenosine 3',5' monophosphate (cAMP) and the media concentrations of progesterone (P) were determined. Although hCG stimulated both cAMP and P formation in CL of all ages, OT alone had no effect on the basal production of these parameters. However, under certain experimental conditions, OT counteracted the stimulatory effect of hCG on both cAMP and P formation. Because the maximal antigonadotropic effect was found for concentrations of OT in the magnitude of the physiologic intraluteal concentration, these in vitro data suggest a role for OT in the luteolytic process of humans. PMID- 3030828 TI - A correlated stereological and functional studies on the long-term effects of ACTH on rat adrenal cortex. AB - The aim of the study was to investigate the effect of prolonged ACTH administration on quantitative structural changes of the rat adrenal cortex and on function of its cells with particular emphasis on correlation of the results of biochemical estimations with stereologic parameters. Daily injections of 20 micrograms ACTH (Synacthen, Depot) for 35 days results in a marked enlargement of the cortex due to an increase in the volume of all the zones. This increase depends upon hypertrophy and hyperplasia of parenchymal cells. At day 21 of experiment the number of parenchymal cells markedly decreased if compared with day 14, the lost of cells being observed mainly in the zona reticularis. Prolonged ACTH treatment only insignificantly changed serum corticosterone concentration and--if calculated per mg of adrenal weight--did not change adrenal corticosterone concentration and 11 beta-hydroxylase activity and decreased corticosterone output by adrenal homogenate. If expressed per adrenocortical cell or per pair of glands, ACTH increased corticosterone concentration and 11 beta hydroxylase activity while the drop in corticosterone output occurred only at days 28 and 35 of experiment. The striking differences in and among various functional parameters depicting adrenal steroidogenesis show for necessity--in case of long-term experiments leading to hypertrophy or atrophy of the gland--of using coupled stereologic and biochemical techniques which better evaluate the cytophysiological state of adrenocortical cells. PMID- 3030829 TI - Histochemical changes in the testes of lead induced experimental rats. AB - The experiments were performed on mature male rats divided in five groups, one control and four experimental in which the animals received 1 mg, 2 mg, 4 mg and 6 mg/kg body weight lead acetate intraperitoneally respectively, over a period of 30 days. ALA-D and lead was estimated in the blood by the use of atomic absorption spectrophotometer and ATP-ase, AMP-ase, Alk-ase were histochemically localized. Significant increase in blood and testis of lead levels along with decrease of ALA-D levels were observed. Changes in the testicular tissue were encountered. Other details concerned with the damage of the testicular tissue are discussed. PMID- 3030830 TI - [Effects of ACTH or hCG on corticosterone and progesterone secretion from adrenal glands of the rats at various ages]. AB - Effects of ACTH or hCG on the secretion of corticosterone (CB) and progesterone (PRG) from adrenal glands of the female rats at various ages were studied. In in vitro experiment, adrenal gland cells were isolated from fetal, neonatal, immature and adult rats by the method described by us before and were incubated in MEM. We chose to designate the hormones in the medium as hormone secretion. In in vivo experiment, hormones in the serum were determined. In vitro experiment, in which ACTH was added to the medium, indicated that CB was low in the media containing the cells from fetal and neonatal glands compared to those from immature and adult. PRG was elevated by ACTH addition at all of the ages. hCG addition had no effects on both CB and PRG in the media of immature and adult rats. In vivo experiment, in which ACTH was injected s.c., indicated that the response of serum PRG was slightly earlier than the CB response and peaked with a subsequent decline in immature rats. The results suggest that adrenal gland responds to ACTH from fetal age and secrets PRG, but little of CB in fetal age. PMID- 3030831 TI - [Adenylate cyclase system responsive to thyroid stimulating hormone (TSH) of porcine thyroid cells in primary monolayer cultures. Potential effect of forskolin on TSH-mediated adenylate cyclase stimulation]. AB - The TSH-responsive adenylate cyclase system was studied using porcine thyroid cells in a primary monolayer culture. Isolated porcine thyroid cells treated with collagenase were inoculated into 96 wells at the density of 5 X 10(4) viable cells/0.25 ml Ham F-12 containing 10% fetal bovine serum and cultured for 4 days in a humidified atmosphere with 5% CO2. Adenylate cyclase activities in the cells treated or non-treated with protein synthesis inhibitor were assayed in Hanks/20 mM Hepes buffer (pH 7.4) containing 1% BSA, 1 mM IBMX and various stimulants at 37 degrees C for 30 or 60 min. The reaction was stopped by adding ice-cold TCA, and cAMP content in the extract was measured by radioimmunoassay after treatment with water-saturated ether. The cultured thyroid cells had an adenylate cyclase system responsive to TSH, cholera toxin and forskolin. TSH (50 mU/ml) stimulated the activity about eight fold over the basal activity. Cholera toxin (1 microgram/ml) and forskolin (100 microM), however, were much stronger activators of the adenylate cyclase system. In the cells pretreated with cyclo-heximide (5 micrograms/ml) up to 24 hours, cAMP formation by TSH was potentiated 200 approximately 170% compared to that in non-treated cells, suggesting a suppression of an inhibitory mechanism dependent upon new protein synthesis. In contrast, forskolin (100 microM)-stimulation was greatly reduced to 30% of the control after 24-hour treatment. Cholera toxin (1 microgram/ml)-stimulation was significantly lessened or slightly reduced by the treatment. Although the ability of forskolin to act synergistically with TSH or cholera toxin was observed in non treated cells, it was clearly unaffected and demonstrated in the cells treated with protein synthesis inhibitor. The mechanism(s) and site(s) of forskolin action still remain unclear. However, these observations are compatible with a two-site model of forskolin action. The direct activating site of forskolin appears to reside in a protein which is closely associated with the catalytic unit of adenylate cyclase system and has a relatively shorter half-life than other components of the system. The potential action of forskolin may reside in a more stable complex of an activated stimulatory guanine nucleotide binding component and catalytic unit of the adenylate cyclase system. Based on these results, it is likely that the primary monolayer culture of porcine thyroid cells is a good model to investigate the adenylate cyclase system in the thyroid, and that forskolin may potentiate the TSH-mediated stimulation of adenylate cyclase. PMID- 3030832 TI - Serum angiotensin-converting enzyme activity in psoriasis. AB - Conflicting data concerning the presence of enhanced serum angiotensin-converting enzyme (SACE) activity in psoriasis are reported in the literature. In order to verify whether this abnormality is a typical pattern of psoriasis, SACE levels were determined in 27 psoriatics as well as in 18 healthy subjects and in 30 patients with essential hypertension. We found that SACE levels were normal in patients affected by psoriasis independently of the presence or the absence of abnormalities in carbohydrate metabolism. Furthermore, SACE levels were not related to the severity of the skin lesions. We conclude that an elevated SACE activity is not a typical pattern of psoriasis, whereas, when present in a psoriatic, it might suggest the possibility of a coexisting unknown systemic sarcoidosis. PMID- 3030833 TI - Structure and function of the mitochondrial ubiquinol: cytochrome c reductase and NADH: ubiquinone reductase. PMID- 3030835 TI - Structure and genes of ATP synthase. PMID- 3030834 TI - The molecular aetiology of human mitochondrial myopathies. PMID- 3030836 TI - Structure and biology of central nervous system neurotransmitter receptors. PMID- 3030837 TI - mRNA-directed synthesis and insertion of functional amino acid receptors in Xenopus laevis oocytes. PMID- 3030838 TI - The control of floral pigmentation in Antirrhinum. PMID- 3030839 TI - Glucagon stimulates adenylate cyclase through GR2 glucagon receptors: a process which can be attenuated by glucagon stimulating inositol phospholipid metabolism through GR1 glucagon receptors. PMID- 3030840 TI - Regulation of signal transduction by signal-controlled effector systems. PMID- 3030841 TI - The dual regulation of cyclic AMP and calcium by luteinizing hormone and the mechanisms of desensitization and recycling of the luteinizing hormone receptor. PMID- 3030842 TI - Inositol trisphosphate and tetrakisphosphate phosphomonoesterases of rat liver. PMID- 3030843 TI - Mapping the binding sites of transducin by sequence homology, chemical modification, and by using monoclonal antibodies. PMID- 3030844 TI - The gamma-aminobutyric acid receptor of Ascaris as a target for anthelmintics. PMID- 3030845 TI - Lipid-protein interactions in the adaptive regulation of membrane function. PMID- 3030846 TI - Translocation intermediates on the import pathway of proteins into mitochondria. PMID- 3030847 TI - Mullerian inhibiting substance blocks autophosphorylation of the EGF receptor by inhibiting tyrosine kinase. AB - The fetal regressor Mullerian inhibiting substance (MIS), in concentrations as low as picomolar, inhibited the growth of A-431 cells and the autophosphorylation of its epidermal growth factor (EGF) receptor. The inhibition of membrane phosphorylation was due neither to the reduction of the total number of EGF receptor binding sites, nor to stimulation of intrinsic phosphates, but rather to inhibition of tyrosine kinase activity. MIS control of EGF receptor autophosphorylation by tyrosine kinase may be one mechanism by which Mullerian duct regression in the embryo and the inhibition of A-431 proliferation is initiated. In addition, MIS as an inhibitor of phosphorylation may furnish a tool to probe the role of membrane phosphorylation in growth control. PMID- 3030848 TI - Dedifferentiation of cultured thyroid cells by epidermal growth factor: some insights into the mechanism. AB - Epidermal growth factor (EGF) has been shown to enhance both the proliferation and dedifferentiation of thyroid cells in culture, leading to a maintained dedifferentiated state, even in the presence of thyrotropin (TSH). Since this maintained loss of differentiated function is not seen with other mitogens, it may relate to a regulatory role for EGF in thyroid function. Therefore, we have examined the loci affected by the dedifferentiative actions of EGF using porcine thyroid cells in culture. EGF (10 ng/ml) induces a loss of thyrotropin (TSH) receptors with a time course identical to the loss in ability to transport iodide. This could account for the difference in extent of iodide uptake and morphological dedifferentiation seen between TSH- and cAMP-supported cells, although the fact that cAMP-supported cells also dedifferentiate implies a lesion distal to the cyclase. Reciprocal plot analysis of iodide uptake in control and EGF-treated cells shows that EGF increases the Km for iodide transport, corresponding to a decreased affinity of iodide pump sites for iodide. These effects on iodide pump affinity and TSH receptor number may result from reversal of thyroid cell polarity in monolayer culture, or they may be the result of more specific actions of EGF at these loci. It has been possible to discriminate between the proliferative and dedifferentiating actions of EGF using amiloride, a non-specific inhibitor of the Na+/H+ antiporter. An optimum concentration of amiloride (0.1 mM) was able to block EGF-stimulated incorporation of [3H]thymidine into DNA without preventing the blockade of iodide uptake, which implies that dedifferentiation is not a consequence of proliferation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3030849 TI - A Leydig cell stimulatory factor produced by human testicular tubules. AB - It is demonstrated that tubular fragments derived from human testes and cultured in vitro produce a factor that stimulates the production of testosterone by human interstitial cells and by Percoll-purified Leydig cells from rat and mouse origin. The active principle in the conditioned media is a thermo-labile and trypsin-sensitive protein with an MW greater than 10,000. The factor is active in the presence as well as in the absence of maximally effective concentrations of LH and its activity is not accompanied by measurable changes in cAMP production. There are several points of analogy between this factor and a Leydig cell stimulatory protein produced by rat Sertoli cells. Molecular weight fractionation of spent media from human testicular tubules using an Amicon ultrafiltration system results in a 38- to 102-fold increase in Leydig cell stimulatory activity in a fraction corresponding to a molecular weight of 10,000 up to 30,000. These figures are comparable to those observed after molecular weight fractionation of spent media from rat Sertoli cells. Dose-response curves with partially purified preparations from human and rat origin yield parallel dose-response curves. In rat Sertoli cells as well as in human testicular tubules, the production of the active principle is stimulated by FSH and dibutyryl cAMP. Finally, maximally effective concentrations of the active principles of human and rat origin display no additive effects whereas additive effects are clearly evident with other Leydig cell stimulatory factors such as LHRHa and EGF. The hypothesis is advanced that the Leydig cell stimulatory factors from tubular origin may act as paracrine regulatory molecules responsible for the effects of FSH on Leydig cell function. PMID- 3030850 TI - Characterization of glucagon receptors in liver membranes and isolated hepatocytes during rat ontogenic development. AB - We studied the functional properties of hepatic glucagon receptors during rat development. Glucagon binding to liver membranes and isolated hepatocytes was significantly less in foetuses and weaning rats than in adult animals, reflecting changes in the number of receptors rather than any change in receptor affinity for the hormone. After correcting for the smaller surface area of foetal hepatocytes, glucagon receptor concentrations were still lower in foetuses than in adult rats. The time courses of glucagon association to liver membranes and of glucagon receptor degradation of prenatal and postnatal rats were similar, while inactivation of glucagon by liver membranes was significantly lower in foetal than in adult rats. Also, glucagon-stimulated production of cAMP was smaller in younger rats. These findings suggest that, according to several criteria, glucagon receptors in the foetus are functionally normal and their delayed development may be important for the metabolic processes occurring during the perinatal age. PMID- 3030851 TI - Dynamics of LHRH binding to human term placental cells from normal and anencephalic gestations. AB - In order to improve our knowledge on human placental hCG production, we studied the binding of an LHRH agonist (N-Ac-Pro1,D-Leu6)-LHRH to third trimester intact placental cells from normal and anencephalic fetuses. In normal pregnancies, specific and saturable binding was found for both LHRH and its analogs with two classes of binding sites. Association constants were 4.7 +/- 2.2 (mean +/- SEM) X 10(5) M-1 for the low affinity sites and 1.7 +/- 0.8 X 10(8) M-1 for the higher affinity sites (P less than 0.01), and the estimated number of sites was 1.71 +/- 0.52 nmol/mg of cell protein and 2.79 +/- 0.54 pmol/mg of cell protein, respectively. Preincubation with increasing concentrations of LHRH agonist induced a progressive decrease in specific binding sites and manifested by a reduction in hCG production which paralleled the concentration of the agonist in preincubation buffer. Studies with placental cells from three anencephalic fetuses showed a decreased binding capacity for LHRH and its agonist, when compared to normal trophoblastic cells, as well as a reduced capacity to produce hCG. Our results suggest that mechanisms dependent upon LHRH binding to its receptor are required for placental hCG production in normal pregnancies. Furthermore our investigation suggests a role for the endocrine feto-placental milieu in the manifestation of these placental LHRH binding sites. PMID- 3030853 TI - Hormonal regulation of inhibin production by cultured Sertoli cells. AB - The hormonal regulation of inhibin production by cultured rat Sertoli cells was examined using a specific radioimmunoassay (RIA) which detects the N-terminal portion of the porcine inhibin alpha chain. FSH, but not hCG or prolactin caused a dose-dependent increase in inhibin production (EC50 for FSH = 2.4 ng/ml); both secreted and intracellular levels of inhibin were increased, but the secreted form represented one-half to two-thirds of the total. The FSH-stimulated production of inhibin was augmented by addition of a phosphodiesterase inhibitor, and could be mimicked by cholera toxin, forskolin, or dibutyryl cAMP, all of which are known to increase intracellular cAMP levels. Inclusion of either dihydrotestosterone or estradiol in the cultures had no effect on inhibin production, both in the presence and absence of FSH. Examination of the conditioned media from forskolin-treated Sertoli cells by gel filtration chromatography revealed a single peak of bioactive and immunoreactive inhibin, at a molecular weight of approximately 32,000, similar to that observed for the porcine and bovine follicular fluid inhibins. Thus, FSH activated the cAMP pathway to stimulate Sertoli cell production of inhibin which in turn suppresses pituitary FSH release to form a closed-loop feedback system. PMID- 3030852 TI - Conformational determinants in receptor recognition of peptide hormones: interaction of parathyroid hormone with the glucagon receptor. AB - Receptor binding assays demonstrate that bovine parathyroid hormone (PTH) and human PTH(1-34) can displace [125I]iodoglucagon from binding to its receptor in rat liver plasma membranes. The displacement of [125I]iodoglucagon requires several thousand-fold more bovine PTH or human PTH(1-34) than glucagon. However, the PTH peptides are more effective than secretin, which up to a concentration of 10(-5) M exhibits no ability to displace [125I]iodoglucagon. The greater potency of PTH compared with secretin occurs despite the fact that secretin shows a great deal of sequence homology with glucagon while PTH shows none. We demonstrate by circular dichroism that in the presence of 3 mM SDS glucagon and hPTH(1-34) have similar secondary structure contents, while secretin is more helical. Our results suggest that receptors can recognize gross conformational features of a peptide hormone in addition to interacting with a specific amino acid sequence. The ability of PTH to interact with glucagon receptors can be modulated by incorporation of charged amphiphiles into the plasma membrane. Negatively charged taurodeoxycholic acid increases the binding of the more cationic PTH while positively charged myristyltrimethylammonium bromide decreases this interaction. These effects demonstrate that receptor specificity can be modulated by its lipid environment and that electrostatic interactions between the hormone and the membrane surface can contribute to receptor binding. PMID- 3030854 TI - Hexose metabolism in pancreatic islets. Absence of glucose-6-phosphatase in rat islet cells. AB - Homogenates of either rat or mouse pancreatic islets, pure rat B cells or insulin producing cells of the RINm5F line catalyzed the hydrolysis of D-glucose-6 phosphate. Relative to protein content, the enzymic activity, which was mainly associated with particulate rather than soluble subcellular material, was much lower in endocrine pancreatic cells than in liver. The rat islet enzyme differed from liver glucose-6-phosphate by its lower affinity for D-glucose-6-phosphate, its lower pH optimum, its greater relative efficiency towards L-glycerol-3 phosphate as distinct from D-glucose-6-phosphate, its restricted lability during exposure to pH 5.0, its inability to act as a glucose-6-phosphate:glucose phosphotransferase, and its insensitivity to inhibition by D-glucose. It is concluded that rat islet cells are virtually devoid of true glucose-6-phosphatase activity. PMID- 3030855 TI - Inhibition by phorbol esters and other tumor promoters of the response of the Sertoli cell to FSH: evidence for dual site of action. AB - Several tumor promoters exert their effects by activating a Ca2+-phospholipid dependent protein kinase (protein kinase C). To study the role of this protein kinase in the regulation of Sertoli cell function, we have evaluated the effect of phorbol esters, mezerein, and teleocidin on the response of the Sertoli cell to FSH. Cells were treated for different time intervals with the tumor promoters, and cell response was measured by stimulating the cell with FSH. 12-O Tetradecanoylphorbol 13-acetate (TPA) had no significant effect on basal cAMP production but markedly inhibited the cAMP response to FSH. Significant inhibition of cAMP accumulation was observed after 15 min treatment with 100 nM TPA, and maximal inhibition developed within 1 h. The decrease in cAMP accumulation was dependent on the dose of phorbol ester used, with an estimated ED50 of 10-20 nM TPA. In a manner similar to TPA, mezerein and teleocidin also inhibited the cAMP response of the Sertoli cell, while the phorbol ester 4 alpha phorbol 12,13-didecanoate (4 alpha-PDD), inactive as a tumor promoter and unable to stimulate protein kinase C activity, was devoid of effect. The promoters that inhibited cAMP response also inhibited the FSH-stimulated androgen aromatization. The dose of TPA producing half-maximal inhibition of estrogen accumulation was again 10-20 nM TPA, mezerein, and teleocidin inhibited estrogen accumulation whether FSH, forskolin or cholera toxin was used to stimulate the Sertoli cell. In contrast, only FSH-dependent cAMP accumulation was inhibited by the tumor promoters, while forskolin and cholera toxin stimulations were not affected. These data suggest that tumor promoters which activate protein kinase C act at two sites of the Sertoli cell response. They alter receptor-mediated signal transduction across the membrane and affect steroidogenesis at a site distal to cAMP accumulation. PMID- 3030856 TI - Blockade of rat testicular gonadotropin releasing hormone (GnRH) receptors by infusion of a GnRH antagonist has no major effects of Leydig cell function in vivo. AB - The purpose of this study was to examine the physiological functions of the gonadotropin releasing hormone (GnRH) receptors present in the rat testis. The receptors were blocked in situ by infusing one testis of adult rats for 7 days with 10-100 ng/h of a potent GnRH antagonist (N-Ac-Ala1, D-p-Cl-Phe2, D-Trp3,6 GnRH) using Alzet osmotic minipumps. The contents of the pump were delivered to the testis through a cannula perforating, and fixed, to the tunica albuginea. A plastic cannula alone was attached to the contralateral testis, to act as a control. Infusion of the antagonist resulted in a dose-dependent decrease of testicular GnRH receptors, up to 90%. Some of the antagonist also occupied GnRH receptors in the contralateral testis and pituitary, but these effects were always clearly less than in the infused testis. None of the doses used affected circulating levels of gonadotropins, prolactin (Prl) or testosterone. However, when the endocrine parameters of the two testes were compared, the 100 ng/h dose of the antagonist resulted in a significant (P less than 0.01-0.05) 16-32% decrease in the testicular content of testosterone, and LH, FSH and lactogen receptors. Similar effects (inhibition of the same parameters by 22-42%) were observed when immature (30-day-old) male rats were treated for 1 week with intratesticular infusions of the antagonist. It is inferred from these observations that, in physiological circumstances, testicular GnRH receptors may mediate stimulatory effects of Leydig cell LH and lactogen receptors, and testosterone synthesis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3030857 TI - Differential expression of crystallin genes during development of the rat eye lens. AB - The concentrations of alpha A-, beta B1-, and total gamma-crystallin mRNAs were measured during development of the rat eye lens, using Northern blot and dot blot analysis. After 14 days of fetal growth a sharp increase in the concentration of all three mRNA species was observed. After birth, the concentration of alpha A crystallin transcripts remains high until 6 months of age, the concentration of gamma-crystallin transcripts decreases gradually, while the concentration of beta B1-crystallin transcripts decreases sharply. The composition of the gamma crystallin mRNA pool was determined by measuring the relative amount of the transcripts of each of the six gamma-crystallin genes using primer extension and S1-nuclease mapping. The transcripts of all six genes are found until the third month after birth. Thereafter the transcripts of the gamma 1-1, the gamma 3-1, and gamma 4-1 crystallin genes are no longer detectable. Later on the transcripts of the gamma 2-1 and gamma 2-2 genes also disappear leaving only the transcripts of the gamma 1-2 crystallin gene at the age of 8 months. The concentration of the six different gamma-crystallin mRNAs is thus regulated differentially. PMID- 3030859 TI - Gene regulation during dedifferentiation in Dictyostelium discoideum. AB - During development of Dictyostelium discoideum, cells acquire the capacity to rapidly recapitulate morphogenesis. Therefore, when cells at the loose aggregate stage are disaggregated and challenged to reaggregate, they do so in a tenth of the original time. If loose aggregate cells are disaggregated and resuspended in buffered dextrose solution (erasure medium), they retain the capacity of rapid recapitulation for 80 min, then completely lose this capacity in a single, synchronous step referred to as the "erasure event." The erasure event sets in motion a program of dedifferentiation during which cells lose developmentally acquired characteristics at different times. The erasure event is inhibited by the addition of 10(-4) M cAMP to erasure medium. The synthesis of 33 growth associated polypeptides, the synthesis of 53 development-associated polypeptides, and the level of 2 development-associated RNAs have been monitored during the erasure program and in cultures inhibited from erasing by the addition of 10(-4) M cAMP. Growth-associated polypeptides begin to be resynthesized and development associated polypeptides exhibit dramatic decreases in rate of synthesis at different times throughout the first 240 min in erasure medium. Inhibiting the erasure event with cAMP has no major effect in the resynthesis of the majority of growth-associated polypeptides. Only one growth-associated polypeptide, V28, is completely inhibited by cAMP, suggesting that it may play a role in the erasure process. In contrast, inhibiting the erasure event with cAMP has a marked effect on the synthesis of development-associated polypeptides, causing a dramatic reduction in the rate at which synthesis decreases for 6 polypeptides, and completely inhibits the decrease in the synthetic rate of 8 polypeptides. The two development-associated RNAs, 16G1 and 10C3, exhibit two distinctly different patterns of loss during erasure, but in both cases cAMP added at time zero of the erasure process dramatically retards or inhibits loss. In addition, when cAMP is added just prior to the erasure event, it inhibits the erasure event and stimulates a rapid increase in the level of 16G1 RNA back to the developmental level. The level of 16G1 RNA then remains at this level for at least 400 min. When cAMP is added after the erasure event, it causes a low, transient increase in the level of 16G1 RNA. These results are considered both in relation to the program of erasure, and in relation to the role of cAMP in the expression of developmental genes during the forward program of development. PMID- 3030858 TI - A lineage-specific gene encoding a major matrix protein of the sea urchin embryo spicule. II. Structure of the gene and derived sequence of the protein. AB - A lambda gt11 cDNA clone isolated by use of a polyclonal antispicule matrix protein antiserum is shown in the accompanying paper [S. C. Benson, H. M. Sucov, L. Stephens, E. H. Davidson, and F. Wilt (1987) Dev. Biol. 120, 499-506] to encode a prominent 50-kDa spicule matrix protein (SM50). This clone was used to select homologous genomic recombinants, and the structure of the gene was determined. The SM50 gene occurs once per haploid genome. It contains a single intron located within the 35th codon. A unique transcription initiation site 110 nucleotide pairs prior to the translation start signal was mapped by primer extension. The mRNA is 1895 nucleotides in length, excluding the 3' poly(A) sequence, and contains a single open reading frame 450 codons in length. Though rare in whole embryo RNA the prevalence of the SM50 mRNA is calculated to be about 1% of the total mRNA in skeletogenic mesenchyme cells. The derived peptide sequence indicates a typical N-terminal signal peptide, and an N-linked glycosylation site near the C terminus. About 45% of the length of the protein is included in a domain composed of consecutive approximate repetitions of a 13 amino-acid element, the consensus sequence of which is Trp-Val-Gly-Asp-Asn-Gln Ala-LeuTrp-Val-IleAsp-Asn-GlnPro+ ++-ValGlu. The protein also contains an internal domain unusually rich in proline residues and a very basic C-terminal region. PMID- 3030860 TI - A cholera toxin-sensitive G-protein stimulates exocytosis in sea urchin eggs. AB - To identify guanine nucleotide binding proteins (G-proteins) in sea urchin eggs and to investigate their role in signal transduction at fertilization, we used cholera toxin (CTX) and pertussis toxin (PTX), which catalyze the specific ADP ribosylation of G-proteins. Cell surface complex, consisting of plasma membranes and adhering cortical vesicles, was prepared from eggs of Lytechinus variegatus and incubated with 32P-labeled NAD in the presence of CTX or PTX. CTX catalyzed the ADP-ribosylation of a 47-kDa polypeptide, whereas PTX catalyzed the ADP ribosylation of a 40-kDa polypeptide. Microinjection of approximately 30 micrograms/ml whole CTX or approximately 20 micrograms/ml CTX subunit A into intact eggs caused exocytosis of cortical vesicles. However, if the eggs were first injected with EGTA (0.6-1.4 mM), injection of CTX did not cause exocytosis. Eggs injected with 0.8-2.8 mM cAMP or 1.0-4.0 mM adenosine 3':5' monophosphotioate cyclic Sp-isomer (cAMP-S), a hydrolysis-resistant analog of cAMP, did not undergo exocytosis. These results suggest that a CTX-sensitive G protein is involved in regulating Ca2+ release and exocytosis of cortical vesicles in sea urchin eggs. PMID- 3030861 TI - Evaluation of a new combined inactivated DPT-polio vaccine. AB - A new combined DPT-Poliovirus vaccine (DPTP) has been developed in France. It contains the DPT components mixed with poliovirus antigens, prepared by culture on vero cells on microcarriers, inactivated by formalin and formulated at a level of 40-8-32 D-antigen units for the type 1, 2 and 3 respectively. A clinical evaluation of this vaccine was conducted in Mali (West Africa) among 320 infants 3 to 24 months of age. The infants were randomly assigned to 3 groups. One group received 2 doses of DPTP, a second group 2 doses of DPT and poliovirus vaccine (IPV) given in two separate sites, the third group 2 doses of DPT and oral poliovirus vaccine (OPV). In each group the 2 doses were given 3 months a part. No severe adverse reactions were observed and no statistical difference was noticed between the three groups for minor side effects. A seroconversion rate of 100% was observed to poliovirus antigens in the groups who received IPV simultaneously or combined with DPT, as compared to 49 to 77% in the group who received OPV. A seroconversion rate of 100%, 100% and 92% was shown in the DPTP group respectively to diphtheria, tetanus and pertussis antigens. No statistical difference was recorded between the 3 groups in the serological response to DPT antigens. A follow-up could be performed in 74 children one year after primary vaccination and a dramatic response to a booster dose of DPTP was measured.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3030862 TI - Safety and efficacy of a combined parapox/BVD vaccine. AB - The combination of a Bovine Viral Diarrhea-Mucosal Disease-attenuated vaccine strain with heat-inactivated parapoxvirus was tested with respect to safety and efficacy in mice, rabbits and fattening calves. The safety on this vaccine was guaranteed in mice and rabbits under laboratory conditions and in calves under field conditions. Laboratory investigations demonstrated a strong anti-BVD antibody response in rabbits compared to antibody formation after vaccination with the monovalent BVD-vaccine component. In mice the induction of high unspecific defence against heterologous viral challenge (VSV, Aujeszky) was possible only with the combined vaccine and the parapox (ORF) vaccine component. Two BVD-seronegative calves kept under isolated conditions showed high titers of BVD-neutralizing antibodies three weeks after a single vaccination with a three fold vaccine dose. Leukopenia could not be detected in 13 randomly chosen BVD seronegative calves on days 1, 2 and 4 after vaccination. In field trials each of the 69 (29%) BVD-seronegative calves developed high titers of BVD-neutralizing antibodies three weeks after a single (i.m.) inoculation. The same phenomenon occurred with animals which had low pre-vaccination BVD neutralizing antibodies, whereas the third group of calves with high prevaccination antibody titers had a moderate decrease in neutralizing antibodies within three weeks after vaccination. Mortality and morbidity rates in fattening farms decreased markedly after the experiment and during an observation period of one year. Improved immunological response of rabbits and cattle to a single vaccination with the combined parapox-BVD vaccine compared to a monospecific BVD vaccination was a result of immunomodulating effects of the parapox (ORF) vaccine component. PMID- 3030863 TI - Evaluation of a combined measles-mumps-rubella-chickenpox vaccine. AB - There was neither a difference in the (low) percentage of only harmless side reactions nor in the rate of sero-conversions (between 95-100%), when groups of 15 to 24 months old children were vaccinated either with the single components or with a combined MMR + Chickenpox-vaccine of a high potency. PMID- 3030864 TI - The combination measles, mumps, rubella and varicella vaccine in healthy children. AB - A clinical trial was conducted to compare the combination measles, mumps, rubella, varicella vaccine (MMRV) and the standard measles, mumps, rubella vaccine and subsequent varicella vaccine (MMR + V) in 15 to 17 month old healthy children. Both the MMRV and MMR + V schedules stimulated virtually 100% seroconversion for all component viruses. Mean antibody titers were similar for each virus component in the two vaccine groups. Clinical reactivity post immunization was also similar with 25-29% morbilliform rashes, 12-25% mild papulovesicular (varicella) rashes, and 12.5-18% temperature elevations above 101 degrees F. Antibodies to measles, mumps, and rubella viruses were persistent in 8/10 originally seronegative MMRV vaccinees and 5/5 MMR + V recipients tested. On MMRV recipient had a household exposure to chickenpox during the year postvaccination that resulted in a subclinical boost in varicella antibody titer. Two children in the MMR + V group had close varicella exposures: one developed mild varicella (20 lesions). There were no known exposures to natural measles, mumps, or rubella. Three of four MMRV vaccinees with low titer antibody to varicella prior to immunization had greater than four-fold rises in antibodies. The combination measles, mumps, rubella, varicella vaccine is an immunogenic, safe and cost effective approach to varicella immunization of healthy children. PMID- 3030865 TI - Protective effect of gamma-linolenic acid on aspirin-induced gastric hemorrhage in rats. AB - The effects of feeding with gamma-linolenic acid (GLA) in comparison with linoleic acid on aspirin-induced gastric hemorrhage were studied in the rat. Gastric damage was examined macroscopically and histologically. Intragastric administration of 100 mg aspirin daily for 4 weeks produced hemorrhage in 3 of 8 rats receiving a linoleic-acid-enriched diet, but none in 8 rats receiving GLA enriched diet. The levels of linoleic acid in plasma and liver phospholipids were significantly increased, whereas those of arachidonic acid (AA) were reduced in plasma and liver phospholipids of aspirin-treated animals fed linoleic acid. Similar, more pronounced changes occurred in those animals with hemorrhage. The reduced ratios of arachidonate/linoleate suggest that fatty acid desaturation in these animals was depressed. Treatment with GLA prevented these changes. Our results demonstrated that GLA could protect the gastric mucosa from aspirin induced damage by bypassing the depressed delta-6-desaturation and thus providing a precursor for the synthesis of AA and prostaglandins. PMID- 3030866 TI - Four factors that accurately predict hearing loss in "high risk" neonates. AB - Two simple overall measures of health--length of stay in the Intensive Care Nursery (ICN) and gestational age--predict hearing loss in ICN graduates. Craniofacial anomalies, congenital perinatal infections, and meconium aspiration are strong predictors of hearing loss, especially in term infants. Findings are based on univariate and multivariate analyses of a number of variables that might be associated with permanent hearing loss. Study variables included all seven High Risk Register items and a number of other features of the ICN history. They were examined in 799 ICN graduates whose hearing had been monitored in their first few years of life. These babies composed 40% of the ICN population and were selected because they had one or more "high risk" factors in their neonatal history. Prevalence of hearing loss in this high risk sample was similar to that found in other ICN samples. Prevalence of hearing loss associated with individual Risk Register items was similar to other published findings for some items and not for others. PMID- 3030867 TI - Effect of various polybrominated biphenyls on cell-cell communication in cultured human teratocarcinoma cells. AB - Firemaster BP-6 (FM), a mixture of polybrominated biphenyls (PBB), and the congeners 2,2',4,4',5,5'-hexabromobiphenyl (245-HBB), 3,3',4,4',5,5' hexabromobiphenyl (345-HBB), and 3,3',4,4'-tetrabromobiphenyl (34-TBB) were tested for their ability to inhibit cell-cell communication between human teratocarcinoma cells in culture. Both FM and 245-HBB were capable of inhibiting cell-cell communication in these cells, with only slight effects on cell survival. 345-HBB was highly cytotoxic, but did not show the ability to interrupt cell-cell communication. 34-TBB was moderately cytotoxic and was also ineffective at blocking cell-cell communication. The results agree with previously published findings in Chinese hamster V79 cells, and are in accordance with proposed structure-activity relationships for these compounds. Inhibition of cell-cell communication, either directly (FM and 245-HBB) or indirectly through necrosis induced compensatory hyperplasia (345-HBB), may be involved in the promotion of hepatocellular neoplasms by these compounds. PMID- 3030868 TI - Comparative toxicology of tetrachlorobiphenyls in mink and rats. II. Pathology. AB - Young female pastel mink and young female Sprague-Dawley rats were injected intraperitoneally on 3 sequential days with 50 mg/kg of either 2,4,2',4' tetrachlorobiphenyl (TCB) or 3,4,3',4'-TCB and sacrificed after 7 days. Two control groups were established for each species; one allowed free access to food, and one pair-fed to the 3,4,3',4'-TCB-treatment group. Heart blood was collected from each mink immediately after sacrifice. A complete set of tissues was collected from all animals and placed in buffered formalin. The rats displayed no clinical signs of illness following the administration of either congener, nor were there any significant gross or microscopic lesions created in this species. Mink in the 2,4,2',4'-TCB and control groups remained free of clinical signs and significant gross or microscopic lesions. Mink in the 3,4,3',4'-TCB group developed anorexia within 48 hr after the initial injection, and depression and melena by Day 4. Necropsy on Day 7 revealed a severe necrotizing enteritis with moderate to marked villus atrophy and fusion in the small intestines of all mink in this treatment group. The epithelial necrosis generally spared the basal one-third of the mucosa, and the deep crypt epithelium was often moderately hyperplastic. The mechanism by which 3,4,3',4'-TCB causes this unique lesion is unknown. PMID- 3030869 TI - [Evaluation of anti-HAV IgG on 2 samples of closed populations]. AB - The authors have carried out an epidemiologic research about the diffusion of antibody to hepatitis A antigen in the inhabitants of Ginostra, fraction of Stromboli, and Alicudi Islands (Eolie's arcipelago). We have examined by ELISA 86 human sera. We have detected a percentage of positivity about 82 and 70.6% for two populations. A critic examination of the results with parameters auxiliary (sex, age), showed significant differences of positivity about two different sexs and ages. PMID- 3030870 TI - [Use of ACE inhibitors in the treatment of cardiac failure and arterial hypertension]. PMID- 3030872 TI - Effects of leukotrienes on gastric acid and alkaline secretions. AB - This study was designed to determine the influence of leukotriene C4 on gastric acid and alkaline secretion. Leukotriene C4 was found to be a potent inhibitor of gastric acid secretion induced in vagally innervated and denervated portions of the stomach of conscious dogs by a variety of stimulants such as histamine, pentagastrin, and meat feeding. Leukotriene C4 was also an effective inhibitor of acid formation in the isolated gastric glands stimulated by histamine or dibutyryl cyclic adenosine monophosphate without or with addition of indomethacin, indicating that this compound acts directly on the parietal cells without mediation of endogenous prostaglandins. Leukotriene C4 was also an effective stimulant of gastric alkaline secretion. However, this effect was probably mediated by an increase in the generation of endogenous prostaglandin, as it was accompanied by an increase in the luminal release of prostaglandin E2 and indomethacin prevented both the stimulation of alkaline secretion and luminal prostaglandin E2 release by leukotriene C4. PMID- 3030873 TI - Unidirectional interaction between thyrotropin releasing hormone and opiates at the level of their brain receptors. AB - The effect of TRH and its analogs, MK771 and RX-77368 on the binding of [3H]D serine-6-threonine-leucine enkephalin (delta opiate receptors), [3H]ethylketocyclazocine (kappa receptors) and [3H]naltrexone (mu receptors) to the rat brain membranes was determined. Neither TRH nor its analogs affected the binding of ligands for mu, delta, and kappa opiate receptors at either 35 or 0 degrees C. Since previous studies from this laboratory indicate that the drugs acting at the kappa, and delta opiate receptors inhibit the binding of [3H](3 MeHis2) TRH to brain TRH receptors, the present studies suggest a unidirectional interaction between opiates and TRH in the brain at the level of their receptors. PMID- 3030871 TI - Steady state regulation of intracellular pH in isolated rabbit gastric glands. Roles for Na/H and Cl/OH (HCO3) exchange. AB - Intracellular pH (pHi) was measured using the fluorescent dye 2'7' bis(carboxyethyl)-5(6)-carboxyfluorescein (BCECF) in cells of isolated rabbit gastric glands that were either suspended in a stirred cuvet or attached to a cover slip. Glands were treated with HCO3-free Ringer's (100% O2 gassing) of different ionic composition and extracellular pH (pHo). In NaCl Ringer's steady state pHi = 7.15 was independent of pHo over the range pHo 7.1-7.8. This regulatory range of pHi (= 0.7 pH units) was abolished by sodium-free Ringer's or addition of 10(-3) M amiloride and also by 10(-4) M ouabain. Chloride-free Ringer's or 2 X 10(-4) M SITS caused an elevation of pHi to 7.5, and the regulatory range was reduced in magnitude to 0.50 pH units (pHo 7.3-7.8). At pHo values outside the regulatory ranges, pHi was linearly related to pHo, although the specific slope that related pHi to pHo depended on the composition of the external solution. In potassium gluconate Ringer's, pHi was completely independent of pHo. The data suggest that Na/H exchange is primarily responsible for the regulation of pHi over a wide range of pHo, although the specific pHi and the range of regulation are affected by the activity of the Cl/OH (HCO3) exchanger; the movement of protons or hydroxyl ions, or both, across gland cell membranes appears to be mediated under most circumstances by these two exchangers. PMID- 3030874 TI - Trifluoperazine inhibits progesterone and cyclic AMP production in granulosa cells of the hen (Gallus domesticus). AB - Unstimulated (basal) as well as luteinizing hormone (LH)-promoted progesterone production in collagenase-dispersed hen granulosa cells was inhibited in a dose related manner by two phenothiazines, trifluoperazine (TFP) and chlorpromazine (CP), both of which are known calmodulin antagonists. Using TFP, the more potent antagonist of the two, it was found that LH-stimulated cyclic AMP production was also suppressed. Moreover, TFP attenuated the steroidogenic effects of both 8 bromo-cyclic AMP and isobutylmethylxanthine but had no effect on the conversion of pregnenolone to progesterone. The inhibitory effects of TFP on steroidogenesis were reversible. It is concluded that phenothiazines inhibit steroidogenesis in ovarian granulosa cells by acting at multiple sites both proximal and distal to cyclic AMP generation without influencing the enzyme complex responsible for the conversion of pregnenolone to progesterone. The results are discussed in relation to calmodulin- and non-calmodulin-mediated actions of phenothiazines. PMID- 3030875 TI - Independence of the pituitary-interrenal axis and melanotroph activity in the brown trout, Salmo trutta L., under conditions of environmental stress. AB - Adaptation of brown trout to a black or white background had no statistically significant influence on the activity of the pituitary-interrenal axis in unstressed fish, in fish under conditions of mild chronic stress, or in fish under conditions of severe, acute stress. This finding is in contrast to several other studies on salmonid fish in which background color adaptation has been shown to modulate the pituitary-interrenal response to environmental stress. It is argued that these differences in results may be related to the prior history of interrenal activation in the experimental fish and that in previously unstressed brown trout the activity of the pituitary-interrenal axis is independent of melanotroph activity in the pars intermedia when fish are exposed to subsequent stress. PMID- 3030876 TI - On the use of dexamethasone to block the pituitary-interrenal axis in the brown trout, Salmo trutta L. AB - Oral administration of dexamethasone in a single meal at a dose of 3 micrograms g 1 body wt was sufficient to block the secretion of ACTH and cortisol in brown trout subjected to a handling stress followed by 30 min confinement. This dose, however, also had a marked effect on several of the circulating blood cell types. Lymphocyte and neutrophil counts were reduced in dexamethasone-treated fish whereas erythrocyte counts increased. The value of dexamethasone as a tool for investigating the role of interrenal tissue during stress responses in teleost fish is limited by its cortisol-like effects on other steroid-sensitive tissues. PMID- 3030877 TI - Effect of dexamethasone and ACTH on oocyte growth and recruitment in the frog Rana cyanophlyctis during the prebreeding vitellogenic phase. AB - The effects of 5, 25, 50 and 75 micrograms of dexamethasone and 0.1 or 0.5 IU ACTH on oocyte growth and recruitment were studied in Rana cyanophlyctis during prebreeding vitellogenic phase (May). Injections (ip) were given 6 days a week for 31 days and frogs were killed on the 32nd day. Treatment with 5 micrograms dexamethasone had no effect on gonadosomatic index (GSI) or on the number and percentage of different oocytes. Administration of 25 micrograms dexamethasone caused a significant (P less than 0.05) increase in both number and percentage of medium second growth phase (MSGP) oocytes and atretic follicles (AF), and a numerical reduction in number and percentage of large second growth phase (LSGP) oocytes. Frogs which received 50 or 75 micrograms dexamethasone exhibited a significant (P less than 0.05) decrease in GSI, ovarian weight, and number and percentage of LSGP oocytes, while those of MSGP oocytes and AF increased. There was no effect of dexamethasone on first growth phase (FGP) oocytes. The administration of 0.1 IU ACTH had no effect on GSI, percentage of different oocytes, or MSGP and LSGP number. There was a numerical increase in number of FGP oocytes. Treatment with 0.5 IU ACTH caused a significant (P less than 0.05) decrease in GSI, ovary weight, and number and percentage of LSGP oocytes, while those of AF increased. The above findings suggest that lower doses of dexamethasone (5 or 25 micrograms) and of ACTH (0.1 IU) have no effect on oocyte recruitment and growth, but that higher doses (50 or 75 micrograms dexamethasone and 0.5 IU ACTH) impair vitellogenic growth of oocytes and increase follicular atresia.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3030878 TI - Germinal vesicle breakdown in oocytes of catfish, Mystus vittatus (Bloch): in vitro effectiveness of mammalian pituitary hormones. AB - The effect of six mammalian pituitary hormones on germinal vesicle breakdown (GVBD) in Mystus vittatus oocytes was investigated in vitro. bLH was found to be most effective at the concentration of 10 micrograms/ml of incubation medium, causing GVBD in 68.4 +/- 1.2% oocytes (P less than 0.001). It could also induce GVBD at the concentrations of 1.0 micrograms/ml (P less than 0.001) and 0.1 micrograms/ml (P less than 0.005). Among other hormones, oFSH could cause GVBD in 16.6 +/- 2.8 and 9.7 +/- 1.0% oocytes at the concentrations of 10 and 1.0 microgram/ml, respectively (P less than 0.05). However, bTSH, bGH, oPRL, and hACTH (synthetic) had no effect on GVBD in the oocytes of this species. PMID- 3030879 TI - Hybrid dysgenesis in Drosophila melanogaster: evidence from sterility and southern hybridization tests that P cytotype is not maintained in the absence of chromosomal P factors. AB - A two-generation crossing program was used to replace the entire chromosome complement of P strains by M strain chromosomes, the maternal contribution being from the P strain. The cytotype of chromosomally substituted females was indistinguishable from M strain cytotype, judged by the sterility of offspring from the cross of such females to P strain males. In addition, following replacement of the chromosomes, the level of DNA homologous to the P factor was sufficiently low to be explicable by low levels of P factor transposition. These results are consistent with immediate chromosomal control for the switching from P to M cytotype. However, the reverse chromosome substitution, replacing all chromosomes of an M strain with P chromosomes, did not usually lead to immediate change of cytotype properties, showing that there is a true maternal effect in the M to P direction. The absence of true maternal inheritance for P cytotype argues against models of P factor repression which depend on autonomous replication of a nonchromosomal element. The repression could still be explained by nonchromosomal copies of the P factor, provided that these are replenished from chromosomal P factors. A model is put forward in which P cytotype is due to the presence of circular P factors carrying a P factor target sequence, leading to preferential transposition of chromosomal P factors to nonchromosomal target sites. PMID- 3030880 TI - On recombination among In(2L)t, alpha-Gpdh and Adh in Drosophila melanogaster. AB - The occurrence and patterns of linkage disequilibrium between an inversion and allozymic loci within it or nearby have been used in attempts to discriminate among different hypotheses for the maintenance of variability. The interpretation of the data on the best-documented case, that of the nonrandom association between In(2L)t and alpha-Gpdh or Adh in the second chromosome of Drosophila melanogaster, has been done on the basis that recombination between alpha-Gpdh and Adh is almost entirely due to the recombination between In(2L)t and the locus within it (alpha-Gpdh), the recombination between the inversion and the nearby locus (Adh) being negligible. In this paper, we show that the pattern of recombination is just the opposite. PMID- 3030881 TI - Sexual hyperactivity and reduced longevity of dunce females of Drosophila melanogaster. AB - The dunce gene of Drosophila melanogaster codes for a cyclic adenosine-3',5' monophosphate-specific phosphodiesterase. Mutations of dunce alter or abolish the activity of this enzyme, produce elevated cAMP levels, cause recessive female sterility, and produce learning deficiencies in both sexes. Aberrant male sexual behavior has also been associated with the memory defects of dunce mutants. Here we show that the longevity of dunce mutant females, homozygous for null-enzyme alleles, is reduced by 50% in the presence of males compared to control dunce females kept without males. Mutant dunce females, mate every 22-24 hr. We propose a cause-effect relationship between mating and reduced longevity. Pheromones or peptides transferred during mating may activate adenylate cyclase and create an increase in cAMP levels that cannot be damped in dunce females. This increase may affect basic physiological functions and lead to reduced longevity. PMID- 3030882 TI - Amplified RNase H activity in Escherichia coli B/r increases sensitivity to ultraviolet radiation. AB - Strains of E. coli B/r transformed with the plasmid pSK760 were found to be sensitized to inactivation by ultraviolet radiation (UV) and to have elevated levels of RNase H activity. Strains transformed with the carrier vector pBR322 or the plasmid pSK762C derived from pSK760 but with an inactivated rnh gene were not sensitized. UV-inactivation data for strains having known defects in DNA repair and transformed with pSK760 suggested an interference by RNase H of postreplication repair: uvrA cells were strongly sensitized, wild-type and uvrA recF cells were moderately sensitized and recA cells were not sensitized; and minimal medium recovery was no longer apparent in sensitized uvrA cells. Biochemical studies showed that post-UV DNA synthesis was sensitized and that the smaller amounts of DNA synthesized after irradiation, while of normal reduced size as indicated by sedimentation position in alkaline sucrose gradients, did not shift to a larger size (more rapidly sedimenting) upon additional incubation. We suggest an excess level of RNase H interferes with reinitiation of DNA synthesis on damaged templates to disturb the normal pattern of daughter strand gaps and thereby to inhibit postreplication repair. PMID- 3030884 TI - Distribution and abundance of insertion sequences among natural isolates of Escherichia coli. AB - A reference collection of 71 natural isolates of Escherichia coli (the ECOR collection) has been studied with respect to the distribution and abundance of transposable insertion sequences using DNA hybridization. The data include 1173 occurrences of six unrelated insertion sequences (IS1, IS2, IS3, IS4, IS5 and IS30). The number of insertion elements per strain, and the sizes of DNA restriction fragments containing them, is highly variable and can be used to discriminate even among closely related strains. The occurrence and abundance of pairs of unrelated insertion sequences are apparently statistically independent, but significant correlations result from stratifications in the reference collection. However, there is a highly significant positive association among the insertion sequences considered in the aggregate. Nine branching process models, which differ in assumptions regarding the regulation of transposition and the effect of copy number on fitness, have been evaluated with regard to their fit of the observed distributions. No single model fits all copy number distributions. The best models incorporate no regulation of transposition and a moderate to strong decrease in fitness with increasing copy number for IS1 and IS5, strong regulation of transposition and a negligible to weak decrease in fitness with increasing copy number for IS3, and less than strong regulation of transposition for IS2, IS4 and IS30. PMID- 3030883 TI - Local DNA sequence control of deletion formation in Escherichia coli plasmid pBR322. AB - The specificity of deletion formation was studied using tests involving reversion of palindromic insertion mutations. Insertions of a Tn5-related transposon at 13 sites in the ampicillin-resistance (amp) gene of plasmid pBR322 were shortened to a nested set of perfect palindromes, 22, 32 and 90 bp long. We monitored frequencies of reversion to Ampr, which is the result of deletion of the palindrome plus one copy of the flanking 9 bp direct repeats (which had been formed by transposition). Revertant frequencies were found to depend on the location and the sequence of the palindromic insert. Changing a 45-kb interrupted palindrome to a 22-bp perfect palindrome stimulated deletion formation by factors of from fourfold to 545-fold among the 13 sites, while elongation of the perfect palindrome from 22 to 90 bp stimulated deletion formation by factors of from eight- to 18,000-fold. We conclude that deletion formation is strongly affected by subtle features of DNA sequence or conformation, both inside and outside the deleted segment, and that these effects may reflect specific interactions of DNA processing proteins with template DNAs. PMID- 3030885 TI - Hsp28stl: a P-element insertion mutation that alters the expression of a heat shock gene in Drosophila melanogaster. AB - We have identified and cloned a mutant allele of the small heat shock gene Hsp28 of Drosophila melanogaster. This allele, which we have called Hsp28stl, produces small amounts of a single aberrantly large, heat-inducible transcript in heat shocked flies, while a normal-sized Hsp28 transcript is present only in fertile females. No Hsp28 transcript at all is detected in mutant prepupae, a stage when wildtype flies show high levels of Hsp28 RNA. We have cloned the Hsp28stl allele, and have found that a 1.3-kb defective P-element is present 5' to Hsp28 in the mutant line. The site of P-element insertion lies between the Hsp28 "TATA box" sequence and the Hsp28 RNA cap site; in contrast to previously described P element insertions, the element at Hsp28stl is flanked by a two base pair duplication of the insertional target sequence. The results suggest that this insert may separate elements regulating heat-inducible and developmental expression of Hsp28, leading to the different patterns of transcription observed. PMID- 3030886 TI - [Effect of promoter duplication in the synthetic interferon gene on the level of its expression]. AB - The expression of the artificial gene for the human leukocyte interferon (IFN) synthesized by the chemo-enzymic method has been studied in Escherichia coli cells. The genetic constructions in which the IFN gene transcription is carried out from A2 and B promoters of T7 phage or from E. coli lacZ UV5 gene promoter have been obtained, based on pEMB101 plasmid. In addition, the level of interferon production, depending on the promoter used, has been studied. It has been shown that the tandemly located promoters increase efficiency of the IFN gene expression. PMID- 3030887 TI - [Induction of unstable mutations in Drosophila melanogaster by microinjection of DNA from oncogenic viruses into the embryo polar plasma. I. Effect of the embryo genotype]. AB - We have studied the spectrum and nature of mutations induced by oncogenic virus DNA injections into wsn, T-007 line of embryos, and those of the first generation hybrids obtained after crossing the T-007 line males with the Oregon R wild line females (hybrid disgenesis). Each line is shown to have a special group of "hot" sites mutating with high frequency under the effect of the oncovirus DNA injected. PMID- 3030888 TI - [Induction of unstable mutations in Drosophila melanogaster by microinjection of DNA from oncogenic viruses into the embryo polar plasma. II. Analysis of the role of injected DNA]. AB - We have demonstrated that the mutagenic effect of oncovirus DNA injected into Drosophila embryos is of two-type locus specificity: the spectrum of mutations induced by the retroviral cDNA (RSV) changed in different recipient stocks and those induced by the adenoviral DNA (Sa7) did not differ in the stocks studied. The Sa7 DNA and the cDNA of RSV induce mutations in different groups of loci. Transpositions of the copia element were found in mutant lines obtained in both cases. PMID- 3030889 TI - [Molecular causes of thalassemia. IV. Cloning of the beta-globin gene in a patient with beta-thalassemia from Azerbaijan and determination of the point mutation in a minor intron]. AB - On the basis of DNA from a beta-thalassemic patient, human gene library has been obtained using bacteriophage lambda EMBL4 as a vector. The recombinants contain human DNA insertions of 15 to 20 kb. The lambda A1 beta 1 clone has been isolated by the method of hybridization of phage plaque replicas to the HhaI fragment of JW102 plasmid containing human beta-globin cDNA. Restriction mapping revealed the presence of a 22 kb human DNA fragment comprising a portion of the beta-globin gene system. Subcloned fragments of beta-globin gene (within the pUC19 plasmid or phage MI3mp10) were sequenced using the Maxam and Gilbert method as well as that of Sanger. 2150 nucleotides in total were analysed. We have detected the point substitution G----A in the 110 nucleotide of minor intron, leading to the formation of an additional splicing site. PMID- 3030890 TI - Specificity of restriction endonucleases and methylases--a review. AB - The properties and sources of all known restriction endonucleases and methylases are listed. The enzymes are cross-indexed (Table I), classified according to their recognition sequence homologies (Table II), and characterized within Table II by the cleavage and methylation positions, the number of recognition sites on the double-stranded DNA of the bacteriophages lambda, phi X174 and M13mp7, the viruses Ad2 and SV40, the plasmids pBR322 and pBR328, and the microorganisms from which they originate. Other tabulated properties of the restriction endonucleases include relaxed specificities (integrated into Table II), the structure of the generated fragment ends (Table III), and the sensitivity to different kinds of DNA methylation (Table V). In Table IV the conversion of two- and four-base 5' protruding ends into new recognition sequences is compiled which is obtained by the fill-in reaction with Klenow fragment of the Escherichia coli DNA polymerase I or additional nuclease S1 treatment followed by ligation of the modified fragment termini [P3]. Interconversion of restriction sites generates novel cloning sites without the need of linkers. This should improve the flexibility of genetic engineering experiments. Table VI classifies the restriction methylases according to the nature of the methylated base(s) within their recognition sequences. This table also comprises restriction endonucleases which are known to be inhibited or activated by the modified nucleotides. The detailed sequences of those overlapping restriction sites are also included which become resistant to cleavage after the sequential action of corresponding restriction methylases and endonucleases [N11, M21]. By this approach large DNA fragments can be generated which is helpful in the construction of genomic libraries. The data given in both Tables IV and VI allow the design of novel sequence specificities. These procedures complement the creation of universal cleavage specificities applying class IIS enzymes and bivalent DNA adapter molecules [P17, S82]. PMID- 3030891 TI - Synthesis of the outer-capsid glycoprotein of the simian rotavirus SA11 in Escherichia coli. AB - The major outer layer protein, VP7, of the simian rotavirus SA11 has been synthesized in Escherichia coli, under the control of the lac promoter, as a fusion polypeptide with beta-galactosidase (beta Gal). The viral protein in the hybrid polypeptide is missing its N-terminal hydrophobic region and 26 amino acids (aa) at its C-terminus; it is flanked at both ends by beta Gal sequences. We have purified the hybrid 145-kDa protein by affinity chromatography using a column specific for beta Gal. Unexpectedly, a second protein of 118-kDa was also specifically bound to the column. N-terminal aa sequence analysis of these two proteins showed that the 145-kDa protein represented the expected fusion product, whereas the 118-kDa protein was apparently the result of initiation of translation at an internal site close to the 3' end of the viral sequence, in the chimeric mRNA. Each of the two polypeptides represented about 2 to 3% of the total protein of the recombinant-plasmid-carrying bacteria. When a bacterial lysate enriched for the hybrid polypeptides was injected into mice, it induced neutralizing antibodies to SA11 rotavirus. PMID- 3030893 TI - Electron microscopic analysis of in vitro transposition intermediates of bacteriophage Mu DNA. AB - Bacteriophage Mu is a highly efficient transposon and the only moveable element for which an in vitro transposition system has been reported. Recently, this system has been used by Craigie and Mizuuchi [Cell 41 (1985) 867-876] to identify and biochemically characterize intermediates in the transposition process. We have utilized the in vitro transposition system to generate intermediates in the transposition process and have analyzed these intermediates by electron microscopic methods. Partial denaturation mapping has shown the intermediates to be theta-shaped structures in which the phi X174 target DNA is joined to the mini Mu plasmid at the ends of the Mu genome. Our results are in agreement with the previous biochemical studies and the type of intermediate we observe is exactly what is predicted by the Shapiro model of transposition [Proc. Natl. Acad. Sci. USA 76 (1979) 1933-1937]. PMID- 3030892 TI - Radio-labeling of the PstI restriction fragments and improvement in the sequencing procedure. AB - A new and simple method has been developed for efficient labeling of the protruding 3' ends of DNA fragments generated by class-II restriction enzymes. This new method utilizes a synthetic oligodeoxynucleotide 5'-GTGCA-3', which is complementary to the protruding 3' end of the fragment and hybridizes with it in such a manner that the end of the restriction fragment will now have a protruding, one-nucleotide (nt)-long 5' end. This then serves as a template to incorporate just one nt at the 3' end of the restriction fragment. This procedure is easy to use, highly efficient (labeling at 80% of the theoretical incorporation) and generates labeled fragments that are suitable for sequencing. We have also improved the nt sequencing procedure of Bencini et al. [Biotechniques 2 (1984) 4-5] by replacing 'A greater than C' reaction with 'G' and 'C' specific reactions and employing butanol precipitation of DNA. PMID- 3030894 TI - Development of an amphotropic, high-titer retrovirus vector expressing the dihydrofolate reductase gene and conferring methotrexate resistance. AB - Altered mouse dihydrofolate reductase gene (DHFRR) was expressed in murine cells using Abelson murine leukemia provirus genome as a prototype vector. A cDNA clone of DHFRR was inserted into a plasmid structure containing retroviral transcriptional as well as packaging signals. The recombinant plasmid was transfected into psi-2 ecotropic cells and the transient virus was used to infect amphotropic PA-12 cells. Recombinant virus (ABL-DHFRR) was detected in the culture medium of transfected PA-12 cells and was free of helper virus. The ABL DHFRR was capable of conferring methotrexate (MTX) resistance to a variety of cells in culture. The titer of ABL-DHFRR virus was at least tenfold higher than other DHFR retroviruses. The ABL-DHFRR virus titer was increased by selection at increasing concentrations of MTX. The presence of the DHFRR in the virus-infected cells was confirmed by assays which showed reduced inhibition of enzyme activity by MTX. A helper-virus-free, amphotropic, high-titer retrovirus containing the altered DHFR was obtained which may be of use as a dominant selectable marker in infecting hematopoietic progenitor cells. PMID- 3030895 TI - Sequence of 3000 nucleotides at the 5' end of Japanese encephalitis virus RNA. AB - The 5' region of the Japanese encephalitis virus (JEV) RNA was cloned and 3000 nucleotides (nt) were determined by sequencing DNA complementary to viral RNA, and genomic RNA, using oligodeoxynucleotide primers and the dideoxy chain termination reaction. Comparison of the nt sequence and the reduced amino-acid sequence of JEV with those of other flaviviruses showed significant homologies, which allowed locations to be assigned for three structural proteins. PMID- 3030897 TI - Physical map of coliphage BF23 DNA. PMID- 3030896 TI - Construction of a Tn5 derivative determining resistance to gentamicin and spectinomycin using a fragment cloned from R1033. AB - A physical and genetic map of the IncP plasmid R1033 was constructed: restriction fragments were subcloned and antibiotic resistance genes were located. The map is consistent with previous reports that R1033 is a derivative of RP4 carrying a 16 kb transposon Tn1696 which contains the antibiotic-resistance determinants present on R1033 but not on RP4. A BamHI fragment from R1033, determining resistance to gentamicin, spectinomycin and streptomycin, was cloned into Tn5, replacing the central Bg/II fragment that determined kanamycin resistance, producing a recombinant transposon Tn5-GmSpSm. This was shown to transpose in Rhizobium leguminosarum at a frequency similar to that of the parental Tn5. PMID- 3030898 TI - [Endoscopic methods of diagnosing and treating adenoido-cystic cancer of the trachea and bronchi]. PMID- 3030899 TI - Lysis of primary hepatic tumours by lymphokine activated killer cells. AB - Lymphokine activated killer cell is a newly described lytic system against a variety of solid tumours and is distinct in several respects from the classic cytolytic T cell and the natural killer systems. This study was conducted to evaluate the lytic activity of lymphokine activated killer cells against fresh autologous and allogeneic, as well as cultured hepatocellular carcinoma cells. Lymphokine activated killer cell was generated by incubating peripheral blood mononuclear cells with various concentrations of recombinant IL-2 (rIL-2, Cetus, USA) for various periods of time. A four hour 51Cr release assay was used to measure cytotoxicity. The results show that fresh and cultured hepatocellular carcinoma cells were only slightly susceptible to natural killer cells. Normal hepatocytes were resistant to lymphokine activated killer-mediated lysis. Lymphokine activated killer cells could be generated from mononuclear cells of hepatocellular carcinoma patients and normal subjects with lytic activity against fresh autologous and allogeneic and cultured hepatocellular carcinoma cells, but lymphokine activated killer cells from the former was less efficient than that from the latter. It is concluded that the adoptive immunotherapy with combined rIL-2 and lymphokine activated killer may be worth trying in early cases of primary hepatocellular carcinoma. PMID- 3030900 TI - Optimum dosage of ispaghula husk in patients with irritable bowel syndrome: correlation of symptom relief with whole gut transit time and stool weight. AB - To determine the optimum dose of ispaghula husk in patients with irritable bowel syndrome (IBS) and to assess the correlation, if any between the relief in patients' symptoms and the whole gut transit time, and the increase in stool weight, a two part study was carried out. In part 1, 14 male patients were given ispaghula husk in increasing doses of 10 g, 20 g, and 30 g a day for a duration of 17 days each (14 days of study period + three days of stool collection). Ten patients completed the trial. The symptom score improved significantly with all the three doses of ispaghula. Both 20 g and 30 g doses of ispaghula were superior to the 10 g dose but there was no significant difference between the 20 g and 30 g doses. There was a significant (p less than 0.001) increase in the daily stool weight with 10 g dose of fibre with further significant increases with the 20 g and 30 g doses. A positive correlation was seen between the improvement in the symptom score and the increase in stool weight with the 10 g dose of ispaghula but not with the 20 g and 30 g doses. Whole gut transit time remained fairly constant throughout the study period and there was no relationship with either the dose of ispaghula, the alteration in stool weight, or the improvement in the patients symptoms. Ten patients completed part 2 of the study in which ispaghula husk was given in the same dose (10 g, 20 g, and 30 g) but in a random order and with a "washout" period of one week between individual doses. Again all the three doses of ispaghula produced a significant improvement in the symptoms; 20 g and 30 g doses were equally effective and both were significantly superior to the 10 g dose. Assessed individually, all the three symptoms improved significantly; improvement in constipation and pain abdomen was more pronounced than diarrhoea. It is concluded that the optimum dose of ispaghula husk in irritable bowel syndrome is 20 g per day. There is some correlation between the increase in stool weight and the improvement in symptom score but the whole gut transit time remains unchanged despite alterations in stool weight and patients' symptoms. PMID- 3030901 TI - Irritable bowel-type symptoms in spontaneous and induced constipation. AB - The prevalence and severity of irritable bowel symptoms was assessed by systematic questioning in 44 constipated volunteers, most of whom had documented slow intestinal transit. All but two had one or more of the following: passage of mucus, rectal dissatisfaction, bloating, and abdominal pain relieved by defecation. All the symptoms were more prevalent than in 17 normal volunteers or in 301 apparently healthy people studied previously. When 12 normal subjects were made constipated with loperamide all developed one or more irritable bowel symptoms. When 24 constipated subjects received effective laxative treatment the prevalence and severity of these symptoms fell markedly. The findings suggest that in some subjects the slowing down of intestinal transit is associated with irritable bowel symptoms. PMID- 3030903 TI - Abnormal metabolism of arachidonic acid in chronic inflammatory bowel disease: enhanced release of leucotriene B4 from activated neutrophils. AB - The metabolism of endogenous arachidonic acid P(AA) was investigated in activated neutrophils from 20 patients with Crohn's disease, 20 with ulcerative colitis, and 25 healthy volunteers. 1-14C-P(AA) was incorporated into intracellular pools of phospholipids prior to activation of the cells with ionophore A23187 and analyses of released arachidonic acid metabolites by thin layer chromatography. Total release of radioactivity expressing the release of arachidonic acid and its metabolites, was equal in the experimental and control groups, which suggests a normal substrate availability. In contrast, there was a marked increase in the relative release of leucotriene B4 (LTB4) and its omega-oxidation products, 20 hydroxy-LTB4 (20-OH-LTB4) and 20-carboxy-LTB4 (20-COOH-LTB4), with LTB4 values exceeding the reference interval in seven of 20 patients with Crohn's disease, median 8.7%, and in six of 20 patients with ulcerative colitis, median 7.7%, as compared with a median of 5.3% in healthy volunteers. Furthermore, a decreased release of unmetabolised arachidonic acid, correlating inversely with the release of LTB4 in all experimental and control groups, and normal values for the production of other metabolites of arachidonic acid--for example, 5 hydroxyeicosatetraenoic acid (5-HETE) and 12-hydroxyheptadecatrienoic acid (HHT), point to an enzymatic abnormality such as increased activity of leucotriene B synthetase. An increased capacity for release of LTB4, the major pro-inflammatory metabolite of arachidonic acid lipoxygenation by polymorphonuclear leucocytes, may contribute to perpetuation of the inflammation and to tissue destruction in chronic inflammatory bowel disease. Our findings agree with previous reports of an increased release of LTB4 by the colonic mucosa in this condition. PMID- 3030902 TI - Effects of an elemental diet, inert bulk and different types of dietary fibre on the response of the intestinal epithelium to refeeding in the rat and relationship to plasma gastrin, enteroglucagon, and PYY concentrations. AB - Refeeding starved rats with an elemental diet resulted in a marked increase in crypt cell production rate (CCPR) in the proximal small intestine but not in the distal regions of the gut. Little effect on CCPR was noted when inert bulk (kaolin) was added to the elemental diet. Addition of a poorly fermentable dietary fibre (purified wood cellulose) had little effect on intestinal epithelial cell proliferation except in the distal colon where it significantly increased CCPR. A more readily fermentable fibre (purified wheat bran) caused a large proliferative response in the proximal, mid, and distal colon and in the distal small intestine. A gel forming fibre only significantly stimulated proliferation in the distal colon; the rats in this group, however, did not eat all the food given. There was no significant correlation between CCPR and plasma gastrin concentrations, but plasma enteroglucagon concentrations were significantly correlated with CCPR in almost all the sites studied. Plasma PYY concentrations also showed some correlation with CCPR, especially in the colon. Thus while inert bulk cannot stimulate colonic epithelial cell proliferation fermentable fibre is capable of stimulating proliferation in the colon, and especially in the distal colon: it can also stimulate proliferation in the distal small intestine and it is likely that plasma enteroglucagon may have a role to play in this process. PMID- 3030905 TI - Malignant fibrous histiocytoma of the uterus. AB - A primary, malignant pleomorphic giant cell tumor of the uterus was studied by light and electron microscopy. The tumor was characterized by spindle cells, plump epithelioid cells, pleomorphic giant cells, osteoclast-like giant cells, and foamy xanthomatous cells. Histochemically, tumor cells did not show either myogenic or epithelial characteristics. Immunohistochemically, tumor cells were devoid of evidence of desmin, cytokeratin, myoglobin, and lysozyme (muramidase), but vimentin was weakly positive, and alpha 1-antichymotrypsin was weakly positive in the cytoplasm of pleomorphic giant cells. Ultrastructurally, tumor cells did not show either myogenic or epithelial features, but they resembled a variant of malignant fibrous histiocytoma. The present case was classified as a storiform-pleomorphic and giant cell type of malignant fibrous histiocytoma of the soft tissues. The uterus is considered to be an additional possible site of malignant fibrous histiocytoma. PMID- 3030906 TI - Association of human papillomavirus infection and vulvar intraepithelial neoplasia: a morphological and immunohistochemical study of 30 cases. AB - Thirty cases of vulvar intraepithelial neoplasia (VIN) were analyzed in order to determine the frequency of association with human papillomavirus (HPV) infection, and the relationship between this association and patient's age, extent of vulvar lesions, and coexistence with cervicovaginal neoplasia. The presence of condyloma or moderate to marked koilocytosis, now considered as morphological evidence of HPV infection, was observed in 66.6% of our cases. A search for HPV antigens, using the peroxidase-antiperoxidase (PAP) method, was performed in 13 selected cases, and positive staining was detected in 3 of them. The presence of HPV infection correlates with a mean age of 48.8 years, 50% of multicentricity of VIN and coexistence with cervical neoplasia in 30% of the cases, as opposed to a mean age of 55.5 years, 10% of multicentricity of VIN and absence of cervical neoplasia in patients without HPV infection. The demonstration of multiple foci of early stromal invasion in a 43-year-old woman, with multicentric VIN lesions associated with HPV infection, indicates that, even in the presence of such clinicopathological features, the risk of developing stromal invasion should be considered. Considerations are made in relation with the presence of HPV antigen in morphological normal epithelium adjacent to the lesion. Therapeutic implications were also investigated. PMID- 3030904 TI - Is bran efficacious in irritable bowel syndrome? A double blind placebo controlled crossover study. AB - Twenty eight patients with classical irritable bowel syndrome completed a double blind placebo controlled crossover trial in which they added to their normal diet a daily supplement of either 12 bran biscuits (1 = 1.3 g fibre) or 12 placebo biscuits (1 = 0.23 g fibre). Each biscuit was given for three months in random order with crossover to the alternative biscuit at three months. After the initial three months therapy, there was a significant symptomatic improvement compared with pretreatment in both the bran treated (p less than 0.01) and placebo treated groups (p less than 0.01), but there was no significant difference in symptom scores between these two groups. There was no further improvement in either group after the second three months treatment with the alternative therapy. When crossover data for all 28 subjects were combined, symptoms scores after three months bran therapy and after three months placebo therapy did not differ significantly. Twenty four patients completed three day stool collections in both treatment periods. When the symptomatic response to bran among 15 subjects in whom stool weights rose on bran was compared with that among nine subjects whose stool weights were static or fell on the bran, it was shown that symptomatic improvement was independent of an increase in stool weight. These data suggest that in irritable bowel syndrome, especially that associated with abdominal pain, the beneficial effects of bran are due to a placebo response which is independent of an increase in stool weight. PMID- 3030907 TI - [Effects of KT1-32 on acute gastric lesions and duodenal ulcers induced in rats]. AB - We studied the effects of KT1-32 (sodium guaiazulene 3-sulfonate) on development of various acute gastric lesions and duodenal ulcers induced in rats. Male Donryu or Sprague-Dawley rats (220-270 g), fasted (but allowed free access to water) for 24 or 48 hr before the experiments, were used. KT1-32 (dissolved in distilled water, 10-100 mg/kg), given p.o. or intraduodenally (i.d.), dose-dependently inhibited the development of gastric lesions induced by HCl X ethanol (60% ethanol in 150 mM HCl), HCl X aspirin (aspirin 100 mg/kg in 150 mM HCl) or aspirin (150 mg/kg in pylorus-ligated preparation) and Shay ulcers (14 hr pylorus ligation). KT1-32 (30 and 100 mg/kg), given p.o. twice (9.5 hr apart), significantly inhibited the development of duodenal ulcers induced by mepirizole (200 mg/kg, s.c.), but did not inhibit gastric lesions developed simultaneously. KT1-32 (30 and 100 mg/kg), given p.o. or i.d., significantly reduced gastric acid secretion when examined using pylorus ligation preparations. KT1-32 (100 mg/kg, i.d.) had no effect on basal and suppressed duodenal HCO3- secretion by mepirizole. These results suggest that KT1-32 is a promising drug for the treatment of gastritis and peptic ulcers. PMID- 3030908 TI - Derivative spectrophotometric studies on cytotoxic effects of carbon monoxide. AB - Characteristics of cytochrome oxidase prepared from hearts of Sprague-Dawley male rats were studied with the use of the fourth derivative spectrophotometry in respect to the cytotoxic effects of carbon monoxide (CO). CO-exposed rats tended to show lower specific activities of cytochrome oxidase than control and recovered rats. Moreover, there was a significant difference in the fourth derivative spectral features of the enzyme: CO-exposed groups indicated peaks at 412 nm in the spectra while controls at 408 nm. This spectral difference seemed to reflect specific effect of CO on cytochrome oxidase, though such trace remained for not more than a day after death. PMID- 3030909 TI - [Neurosarcoidosis: diagnosis and therapy]. AB - Sarcoidosis is an inflammatory systemic disease with a prevalence of 20 to 50 of 100,000 inhabitants in West-Germany. Caused by an abnormal immune response to a hypothetic inhalative antigen, a granulomatous inflammation in mediastinal lymph nodes occurs, which frequently extends to the lung and sometimes to other tissues. The nervous system is clinically involved in 5% of the cases. The most frequent neurologic symptom is solitary or combined cranial nerve involvement. Cerebral manifestations are nodular or diffuse granulomatous infiltration of the brain and lymphocytic inflammation of the basal meninges. The rarer myelopathies and peripheral neuropathies often cause major neurological deficits. Muscular involvement, which is present in about half of the cases of sarcoidosis, normally causes clinically silent lesions. Chronic sarcoid myopathy or acute polymyositis are rare. Elements of the diagnostic procedure are X-ray of the lung, measurement of serum Angiotensin-Converting Enzyme (ACE)- and serum Lysozyme (LZM)-levels, Bronchoalveolar lavage, Gallium-67-scanning and Kveim-test. The diagnostic value of these investigations and the typical findings in CSF examination, computed tomography, nuclear magnetic resonance, myelography and electrophysiological investigation are presented. Suspected diagnosis of sarcoidosis should be confirmed by biopsy of non-caseating granulomatous lesions in involved tissues. An initially high dosage corticosteroid therapy for many months to several years is often effective. If this is not helpful in cerebral sarcoidosis whole brain irradiation with 1500 to 3000 rd administered in 10 single doses is indicated. Peripheral neuropathy refractory to therapy with orally administered cortisone should be treated with parenteral ultrahigh steroid therapy. PMID- 3030910 TI - A comparison of the effects of isosorbide dinitrate on isolated rabbit aorta and vena cava: role of guanosine 3',5'-cyclic monophosphate in vascular smooth muscle relaxation. PMID- 3030911 TI - Crystalline structures in pulmonary cytomegalic inclusion cells from a case of AIDS. PMID- 3030912 TI - [A simple method for high-resolution banding of chromosomes and its application to diagnosis of birth defects]. AB - A simple method for obtaining high-resolution banding patterns on elongated chromosomes is devised as follows: Peripheral lymphocytes cultured for 3 days with phytohemagglutinin were exposed to 10 micrograms/ml ethidium bromide for 2 hours and with 0.02 micrograms/ml colcemid for 1 hour prior to harvesting. After preparation by steam-dry method, high-resolution G-bands were obtained by Giemsa staining following exposure to hydrogen peroxide and trypsin treatment. 19 cases of the Prader-Willi syndrome, 3 cases of the aniridia-Wilms' tumor syndrome, 2 cases of the Langer-Giedion syndrome, 2 cases of retinoblastoma and 6 cases with multiple congenital anomalies whose diagnosis was not made by routine chromosome analysis were examined by this method. 18 of the 19 cases of the Prader-Willi syndrome had a deletion of 15q11.2. 2 of the 3 cases of the aniridia-Wilms' tumor syndrome had a deletion of 11p13 and the other had a balanced insertion with a breakpoint at 11p13. The 2 cases of the Langer-Giedion syndrome had a deletion of 8q23.3-24.13. One of retinoblastoma had a deletion of 13q14 and the other had a balanced reciprocal X/13 translocation with breakpoints at Xp11.21 and 13q12.3. The last 6 cases had subtle chromosomal aberrations: 46, XX, del (5) (q15q22), 46, XX, del (13) (q32.3), 46, XX, inv dup (6) (p25p21.3), 46, XX, -5, +der (5), t (3; 5) (p23; p13) mat, 46, XX, -5, +der (5), t (5; 9) (p13.3p21) mat, and 46, XX, -4, +der (4), t (4; 16) (p15.2; p13.1) pat. The simple method for high-resolution banding of chromosomes is useful for diagnosis of many kinds of birth defects. PMID- 3030913 TI - Spent media from immature seminiferous tubules and Sertoli cells inhibit adult rat Leydig cell aromatase activity. AB - Adult rat Leydig cell aromatase activity is stimulated 2.5 fold by LH or dbcAMP. Spent media prepared from seminiferous tubules or Sertoli cells of immature rats depress both the basal and the LH stimulated estradiol syntheses (25 and 20% decreases, respectively). These inhibitory effects are further enhanced when FSH is added to the culture medium of seminiferous tubules or Sertoli cells. Rat serum as well as culture media from other cell lines are ineffective while seminiferous tubule media from other immature animals (mouse, guinea-pig, calf) inhibit the aromatase activity. This Sertoli cell factor is a heat stable protein (molecular weight greater than 10 kDa), different from the LHRH-like Sertoli cell compound, which acts on the aromatase activity at a step beyond the adenylate cyclase. PMID- 3030914 TI - Angiotensin-converting enzyme activity decreases during fasting. AB - Serum angiotensin-converting enzyme (ACE) activity varies directly with thyroid hormone levels in states of altered thyroid function. Because T3 levels decrease during fasting, ACE activity was examined to ascertain if it was reduced in this low T3 condition. Eighteen obese euthyroid subjects were hospitalized and placed on a weight-maintaining diet for 4 days. Nine subjects (Group 1) underwent a fast (50 kcal/day) for 8 days. Nine (Group 2) subjects received T3 (5 micrograms q 3 h) during an identical fast. Weight loss was identical in both groups (-4.4 kg). Serum T3 fell in Group 1 from 104 +/- 8 to 50 +/- 4 ng/d/(p less than .05) but was unchanged in Group 2 (114 +/- 11 ng/dl fed vs. 120 +/- 14 ng/dl fasted). Blood pressures fell significantly in Group 1 (mean systolic: 112----104 mmHg; diastolic: 71----65 mmHg, p less than 0.05), but not in Group 2 subjects. ACE activity fell progressively in Group 1 subjects during fasting (14.4 +/- 1.6 U/ml fed vs. 12.8 +/- 1.4 U/ml fasted p less than 0.05). ACE activity was not decreased significantly early in the fast in patients given T3, but by late fast (days 6-8) was reduced to the same degree as in Group 1 subjects. Glucose and insulin levels fell similarly in both groups. CONCLUSIONS: (1) ACE activity is reduced during starvation. This effect is not mediated by T3. (2) Blood pressure reduction during fasting may result from the low T3 levels, but not from decreased ACE activity. Interpretation of serum ACE activity must be viewed in the context of a patient's diet. PMID- 3030915 TI - Angiotensin converting enzyme in hyper- and hypothyroid rats. PMID- 3030916 TI - Hereditary defect of hepatobiliary cysteinyl leukotriene elimination in mutant rats with defective hepatic anion excretion. AB - Hepatobiliary and renal elimination of cysteinyl leukotrienes were investigated in a mutant rat strain with a hereditary defect in the hepatobiliary excretion of conjugated bilirubin, dibromosulfophthalein and ouabain. After intravenous injection of [3H]leukotriene C4, the initial half-life of radioactivity circulating in blood was 79 +/- 15 sec (S.D.) in transport mutant rats as compared to 31 +/- 6 sec (S.D.) in normal Wistar rats. The intrahepatic leukotriene radioactivity was increased 5-fold after 1 hr in mutant rats, while the biliary elimination of [3H]leukotrienes was reduced to 1.8% of control. In normal rats, 77 +/- 7% (S.D.) of the administered leukotriene radioactivity were recovered in bile within 1 hr. The total recovery of radioactivity from bile, urine, liver, intestine, stomach, kidneys, muscular system and blood 1 hr after intravenous [3H]leukotriene C4 was 89 +/- 6% (S.D.) in normal rats and 46 +/- 4% (S.D.) in transport mutants. Enterohepatic circulation was studied after intraduodenal administration of N-acetyl-[3H]leukotriene E4, a major cysteinyl leukotriene metabolite in rat bile. In transport mutants, hepatobiliary elimination of the intestinally absorbed [3H]leukotriene was reduced to 5%, whereas urinary excretion was not significantly affected. [3H]Leukotriene metabolites in bile, liver and urine were separated by reversed-phase high performance liquid chromatography. The proportion of N-acetyl-[3H]leukotriene E4 relative to polar leukotriene metabolites was higher in the bile of transport mutants as compared to control Wistar rats when analyzed within 30 to 60 min after intravenous injection of [3H]leukotriene C4.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3030917 TI - Interaction of natural and synthetic albumin polymers with hepatocytes. AB - The hepatitis B virus binds avidly to albumin polymers which in turn may mediate the initial binding of viral particles to the liver cell. However, the interaction of albumin polymers with the liver remains poorly characterized, and the possibility that hepatic binding reflects an artifact of polymerization with glutaraldehyde has not been excluded. We therefore characterized the binding of 125I-labeled natural and synthetic albumin polymers to suspensions of rat hepatocytes. Saturable binding was demonstrated for all preparations of monomeric and polymeric albumin studied. Glutaraldehyde-polymerized albumin (mean polymerization number = 15) bound much more avidly than naturally occurring albumin polymers (mostly dimers and trimers) or monomeric albumin. Competition between monomer and synthetic polymer was not observed. Reduction of free aldehyde groups on the synthetic polymer decreased nonsaturable binding without affecting saturable binding. Autoradiography confirmed binding of polyalbumin to hepatic parenchymal cells. Glutaraldehyde-polymerized ovalbumin, a protein unrelated to serum albumin, also bound hepatocytes saturably. We conclude that hepatic binding of synthetic albumin polymers is not due to residual aldehyde groups on the polymer and is much more avid than for natural polymer. This difference may reflect the higher degree of polymerization or chemical modification of the synthetic polymer. The hepatic binding sites for synthetic polymer appear distinct from those previously described for monomeric albumin and may not be specific for albumin. PMID- 3030918 TI - Cytophotometric measurements of nuclear DNA content in hepatocellular carcinomas. AB - The nuclear DNA contents of 54 hepatocellular carcinomas were measured on cytologic smears using a scanning microdensitometer at a wavelength of 550 nm. The cellular materials were aspirated under ultrasonographic guidance and stained with modified Feulgen method. The DNA modal values, which could be defined in 51 cases, ranged from 2.2 to 9.8 ploidy, with 46.3% (25/54) in hypertetraploidy. The hepatocellular carcinomas in patients with higher serum levels of alpha fetoprotein contained more hyperploid cells. The DNA modal values did not correlate with the age, HBsAg status, tumor size and histological grading of hepatocellular carcinoma. The S-phase fraction values, which could be estimated on the DNA histograms in 43 cases (79.6%), ranged from 1 to 35 (mean +/- S.D. = 10.6 +/- 8.1). The S-phase fraction values were higher in high grade hepatocellular carcinomas (13.4 +/- 8.5) than low grade hepatocellular carcinomas (6.1 +/- 4.4), p less than 0.01. The mean S-phase fraction values were also significantly higher in hepatocellular carcinomas with alpha-fetoprotein levels of more than 100 ng per ml than those of less than 100 ng per ml (12.9 +/- 8.0 and 7.8 +/- 5.6, respectively). The S-phase fraction values also did not correlate with the age, HBsAg status and tumor size. PMID- 3030919 TI - Proaccelerin and prothrombin assays for detection of hepatocellular carcinoma. PMID- 3030920 TI - Glucose-6-phosphatase activity of Reed-Sternberg and Hodgkin cells. AB - The glucose-6-phosphatase (G-6-Pase) enzyme cytochemical reaction was successfully applied on lymph node imprints, after introducing the appropriate modifications related to fixation and to composition of the substrate solutions. Using this method we studied the G-6-Pase cytochemical profile on lymph node imprint preparations from 30 patients with Hodgkin's disease. Three cases had the histologic subtype of lymphocyte predominance, 14 the modular sclerosis, 11 the mixed cellularity and two the lymphocyte depleted. A strong G-6-Pase positivity was found in the Reed-Sternberg (RS) and Hodgkin cells of the mixed cellularity type, while the RS and Hodgkin cells of the other subtypes were less positive. The rest of the cell population present were G-6-Pase negative, with the exception of plasma cells which exhibited a strong G-6-Pase cytoplasmic positivity. We concluded from our study that RS and Hodgkin cells resemblance to plasma cells and therefore they may be of B-cell origin. In addition it appears that RS and Hodgkin cells express different functional properties in the various histologic subtypes of the disease. PMID- 3030921 TI - Acetone/periodate-lysine-paraformaldehyde (PLP) fixation and improved morphology of cryostat sections for immunohistochemistry. AB - Cryostat sections are extensively used in the study of lymphoid tissues because the majority of antigens do not survive routine fixation and processing. A major disadvantage of cryostat section immunohistochemistry is their poor morphology. The use of periodate-lysine-paraformaldehyde (PLP) fixation regimes for cryostat sections was evaluated and compared with conventional acetone immersion. This fixation gives excellent morphology but poor immunostaining. The quality of immunostaining with a panel of 31 antibodies was good with brief acetone fixation followed by PLP fixation. Morphological preservation was very good. We suggest that acetone-PLP fixation is an improvement over acetone alone for cryostat section immunohistochemistry. PMID- 3030922 TI - Malignant rhabdoid tumor of the central nervous system. AB - Originally described and most frequently reported in association with the kidney, the malignant rhabdoid tumor (MRT) is a highly aggressive neoplasm with distinctive morphologic features. Extrarenal sites reported for this neoplasm include the liver, thymus, and various soft tissue sites. Young infants are affected with rare exceptions. We report the case of a 3-month-old boy who presented with hyperirritability and increasing head size over several weeks. The patient died following a two-week hospital stay marked by development of seizures, paralysis, and apnea. At autopsy, significant findings were limited to the central nervous system. The subarachnoid space contained neoplasm throughout, with multiple areas of parenchymal invasion. A predominating intraparenchymal mass was present in the inferior cerebellum contiguous with the neoplasm in the subarachnoid space and probably represented the site of origin. Microscopically, the neoplasm was composed of a highly cellular monomorphic population of polygonal cells with roughly ovoid vesicular nuclei and conspicuous nucleoli. Variable amounts of cytoplasm were present, and many cells contained a single, well-demarcated eosinophilic hyaline globule adjacent to the nucleus. Ultrastructurally, the cytoplasmic globules were composed of whorled aggregates of intermediate filaments. Immunoperoxidase studies confirmed that the filaments were composed, at least in part, of vimentin. The morphologic and immunohistochemical features are diagnostic of MRT, an entity of unknown histogenesis that has not been reported previously as a primary neoplasm of the CNS. PMID- 3030924 TI - An alpha-fucosidase pseudogene on human chromosome 2. AB - In Chinese hamster-human hybrids with overlapping translocations, the major site of hybridization of a cDNA clone for the liver form of human alpha-L-fucosidase was 1p36.13----1p34, consistent with hybridization to the FUCAl locus. No hybridization to the FUCA2 locus on chromosome 6 was observed. Hybridization to a genomic sequence on chromosome 2 was, however, detected, thus defining a new FUCA like locus. The restriction map of the alpha-fucosidase cDNA could be exactly superimposed upon its region of homology within a genomic clone containing this FUCA-like locus, suggesting that it is a processed pseudogene. PMID- 3030923 TI - DNA microextraction from dried blood spots on filter paper blotters: potential applications to newborn screening. AB - Microextraction of DNA from dried blood specimens would ease specimen transport to centralized laboratory facilities for recombinant DNA diagnosis in the same manner as use of dried blood spots allowed the broad application of screening tests to newborn populations. A method is described which reproducibly yields 0.5 microgram DNA from the dried equivalent of 50 microliters whole blood. Though DNA yields decreased with storage of dried specimens at room temperature, good quality DNA was still obtained. Sufficient DNA was routinely obtained for Southern blot analysis using repetitive and unique sequences. This microextraction procedure will allow immediate application of molecular genetic technology to direct newborn screening follow-up of disorders amenable to DNA diagnosis, such as sickle cell anemia, and may eventually permit primary DNA screening for specific mutations. PMID- 3030925 TI - Alpha-thalassemia haplotypes in the Algerian population. AB - DNA mapping was performed in seven unrelated Hb H patients and nine carriers for alpha-thalassemia trait originating from Algeria. This study has allowed us to identify four alpha-thalassemia haplotypes: the (-alpha 3.7) haplotype, which is the most frequent (18 of 23 alpha-thalassemic chromosomes), the (-(alpha)20.5) haplotype, a (--) haplotype, and an (alpha alpha)T haplotype. Our results also show that the (-alpha 3.7) haplotypes encountered in the Algerian population are heterogeneous and differ by the site of the unequal crossover responsible for the 3.7-kb deletion and the size of the interzeta fragment. In addition, during this survey we observed that normal chromosomes bearing a polymorphic BglII site are associated with different interzeta fragments. PMID- 3030926 TI - DNA deletions in mild and severe Becker muscular dystrophy. AB - The DNA of 33 patients diagnosed as suffering from Becker muscular dystrophy (BMD) has been probed with cloned DNA sequences from Xp21, known to reveal DNA deletions in patients suffering from the more severe Duchenne muscular dystrophy (DMD). Two BMD cases showed clear deletions. A third case gave aberrant band sizes, which further analysis showed to be caused by a small deletion. This suggests that deletions in DXS164 occur approximately as frequently in BMD as they do in DMD. Of the two cases showing large deletions, one is at the severe end of the Becker clinical spectrum, whilst the other is a classical Becker-type dystrophy. The fact that loci defined by probes commonly deleted in classical DMD patients are also deleted in BMD patients of varying severity is strong additional evidence that these disorders are allelic, and further justifies the use of probes with defined linkage relationships to DMD also being used for counselling in BMD families. PMID- 3030928 TI - Exercise radionuclide ventriculography (Ex-RNV) in evaluation of coronary artery disease: experience of 3,300 cases. PMID- 3030929 TI - Insulinoma--a review of six cases. PMID- 3030927 TI - Prenatal diagnosis of X-linked choroideremia with mental retardation, associated with a cytologically detectable X-chromosome deletion. AB - We describe a family in which an X-chromosome deletion is segregating with choroideremia, and X-linked recessive condition. The DNA sequences DXYS1 and DXS3, defined by the probes pDP34 and 19.2 respectively, are absent in the affected male (who is also mentally retarded), and hemizygous in his mother and in his carrier sister, who presented early in pregnancy. Analysis of chorionic villus DNA formed the basis of prenatal exclusion of choroideremia in her male fetus. In three female relatives, studied with late-labelling techniques, the deleted X was preferentially inactivated in 86-100% of cells studied. This family confirms the localisation of the choroideremia locus to within Xq13----21, and places the loci for anhidrotic ectodermal dysplasia and the X-linked immunodeficiencies outside this region. PMID- 3030930 TI - [Hand, foot and mouth disease. Study of 25 cases]. PMID- 3030931 TI - Rotavirus vaccine: current status. PMID- 3030932 TI - Kala-azar in Bihar. PMID- 3030933 TI - Chemotherapy of leishmaniasis in India. PMID- 3030934 TI - Effect of para-chlorophenylalanine on male rats: histopathological and biochemical changes in the testes. AB - Spermatogenically active testes of rat challenged by 100 mg/kg body weight of p- Chlorophenylalanine for 45 days displayed marked and drastic changes in the seminiferous epithelium. Degenerative changes followed by immense necrosis of germ cells lead to complete breakdown of seminiferous tubules. Leydig cells, however, remained unaffected histologically in the treated animals. Among the accessory sex organs, epididymis alone showed a marked decrease in its weight. A biochemical study in the drug treated rats revealed a significant accumulation of glycogen in the testes accompanied by increase in the activities of enzymes like the succinic dehydrogenase, glucose-6-phosphatase, ATP-ase and acid phosphatases. However, a marked decrease was noticed in the activities of enzymes like alkaline phosphatase, phosphohexose isomerase and lactate dehydrogenase. No significant change was found in the protein, DNA and RNA concentrations in the drug treated testes. The histological and biochemical changes induced in the testes by p-CPA suggest the deleterious effect of the drug on the seminiferous tubules of the testes. PMID- 3030935 TI - The Bacteroides by-product succinic acid inhibits neutrophil respiratory burst by reducing intracellular pH. AB - Short-chain fatty acids, particularly succinic acid, are major metabolic by products of Bacteroides species. To determine their role as potential virulence factors in infections containing Bacteroides species, short-chain fatty acids were examined for their effect on the neutrophil respiratory burst. Succinate (30 to 50 mM) irreversibly impaired superoxide and hydrogen peroxide production in response to opsonized zymosan and phorbol myristate acetate when neutrophils were treated at pH 5.5 but not pH 6.0 or greater. Several other short-chain fatty acids tested produced similar inhibition. Reversible inhibition of oxygen consumption was found when neutrophils were incubated in succinate-containing medium (pH 6.0) as well as control medium (pH 6.0 and 5.5). Neutrophil cytoplasmic pH was measured by fluorimetric techniques to determine whether the inhibition was mediated via a reduction in intracellular pH. The intracellular pH of cells in control medium (pH 6.5 or less) was significantly reduced compared with pH 7.4. The addition of succinate (30 mM) to these media caused a further significant reduction in cytoplasmic pH at each pH level. The reduction in intracellular pH was sufficient to account for both the irreversible and reversible impairment of the neutrophil respiratory burst. Thus, short-chain fatty acids appear to exert their inhibition, at least in part, by reducing intracellular pH. These data also demonstrate the potential for interactions between Bacteroides species and their microenvironment to increase the virulence of an infection. PMID- 3030936 TI - Dephosphorylation of the lipid A moiety of Escherichia coli lipopolysaccharide by mouse macrophages. AB - An Escherichia coli deep rough lipopolysaccharide (LPS), biosynthetically labeled with 32PO4 and [3H]glucosamine, was used to study dephosphorylation of the lipid A moiety by murine macrophages. Over a 48-h incubation period, the macrophages removed approximately two-thirds of the 32P from [3H32P]LPS that was added to the culture medium. The LPS-derived phosphate was incorporated into cell components (e.g., phospholipids), as well as released from the cells. Cell lysates were also able to remove phosphate from [3H32P]LPS. The phosphatase activity was optimal at acidic pH and was greatly reduced by 10 mM sodium fluoride or heating at 80 degrees C. There was no evident difference in the LPS-dephosphorylating ability of macrophages from LPS-responsive and -hyporesponsive mice. The results indicate that murine macrophages dephosphorylate the lipid A moiety of deep rough E. coli LPS and raise the possibility that enzymatic dephosphorylation may modify LPS bioactivity. PMID- 3030937 TI - Inhibition of human natural killer cell activity by Pseudomonas aeruginosa alkaline protease and elastase. AB - The present study was designed to examine the effect of Pseudomonas aeruginosa alkaline protease (AP) and elastase (Ela) on human natural killer (NK) cell activity in vitro. AP and Ela were found to inhibit NK cell function. Addition of alpha interferon and interleukin-2 did not abolish this inhibition of NK cell activity. Adhesion of effector to target cells was studied in a single-cell agarose assay of monocyte-depleted NK-cell-enriched cell populations. AP and Ela were shown to inhibit effector/target cell conjugate formation. Furthermore, AP and Ela inhibited the binding of the monoclonal antibody Leu-11, which reacts with the Fc receptor of NK cells. The inhibition of NK cell binding to the target cell by P. aeruginosa proteases is most likely due to proteolytic cleavage of the surface receptors involved in the binding of the effector cell to the target cell. PMID- 3030938 TI - Biofiltration with bicarbonate as dialysate buffer. AB - The biofiltration with bicarbonate as dialysate buffer (BiBF) was used in 10 patients on RDT: the patients were treated for 10 months on standard BF and for 10 months on BiBF. The amount of fluid infused varied between 3 and 5 liters and Na-bicarbonate (100 mEq/h) was infused during BF. The dialytic protocol was 3 hours every other day. Cardiovascular stability, waste molecules and acid-base balance were investigated. No differences in vascular stability and no significant changes in the waste-molecules concentrations were found. Both protocols correct the metabolic acidosis; however, in standard BF 50% of patients showed acute hypocapnia at the end of dialysis. PMID- 3030939 TI - Susceptibility of various human tissues to transformation in vivo with human papillomavirus type 11. AB - Human papillomaviruses (HPVs) are associated with proliferative lesions in a variety of human epithelial types. In some cases, the associations are highly specific in that a certain papillomavirus type may infect only one human epithelial cell phenotype. In other cases, one papillomavirus apparently infects several tissue types and not others. Further, HPV replication capacity varies considerably in different epithelia. Due to the lack of a suitable method, it has not been possible, until now, to define the role of the host cell phenotype in determining the outcome of infection with an HPV. We recently developed a system in which human epithelium was exposed to HPV-II and transplanted to athymic mice. Cervix and skin grafts developed into typical condylomata. We now test the hypothesis that the outcome of infection of diverse epithelial types with a single human papillomavirus is largely determined by the phenotype of the original epithelial cell. For example, skin obtained from a number of disparate sites from a single patient varied dramatically in its capacity for morphological transformation with HPV-II. Skin from penis was highly susceptible, but skin from abdomen did not transform. Vocal cord from 3 children and 2 adults responded to HPV-II infection by producing typical laryngeal papillomata. A variety of tissues were obtained from II donor infants and infected with HPV-II. Foreskin and cervical tissues of these children were transformed at a frequency of 100%. Vocal cord was transformed at an incidence of 88% and urethra at 73%. Only 37% of esophagus samples were transformed and both abdominal skin and urinary bladder from the same infants were totally resistant to morphological transformation. In a separate study, ureteral tissues from a child and an adult were completely resistant to HPV-II infection. Papillomavirus replication was readily detected as capsid antigen in foreskin, cervix, and urethra, but was poorly expressed in morphologically-transformed esophagi and vocal cords. In the last two tissues HPV II DNA and RNA were demonstrated in cells by in situ hybridization techniques. We conclude that the epithelial cell phenotype is a major determinant of HPV-II infection, controlling both morphological transformation and viral replication. PMID- 3030940 TI - Characterization of EBV-carrying B-cell populations in healthy seropositive individuals with regard to density, release of transforming virus and spontaneous outgrowth. AB - Peripheral or tonsil lymphocyte populations of EBV-seropositive donors give rise to EBV-carrying LCLs upon in vitro explantation. Such lines can arise either by a 2-step mechanism, namely release of virus from some of the explanted cells followed by infection of previously uninfected B cells, or by direct outgrowth of virus-harboring B cells (Rickinson et al., 1974; Dalens et al., 1975; Hinuma and Katsuki 1978; Katsuki et al., 1979). We observed that cells responsible for both the 2-step mechanism and for direct outgrowth are found in the purified B-cell compartment. Virus release was more frequent than direct outgrowth. The majority of virus-releasing cells were found in the low-density fraction that contains large, activated B blasts. Cells that were capable of spontaneous outgrowth in the presence of the viral inhibitor PFA and of virus-neutralizing antibody gave rise to cell lines that carried the sex chromosome marker of the original donor, rather than that of admixed cord blood lymphocyte of the opposite sex. Such cells were found in both the low- and the high-density fractions. The majority of the EBV-carrying B cells in vivo are thus low-density blasts. Rare small B cells of high density harboring EBV were capable of spontaneous outgrowth. This may be indicative of a host control mechanism that is removed upon cultivation in vitro. PMID- 3030941 TI - Expression of multiple growth factors in a human lung cancer cell line. AB - U-1810, a human large-cell lung cancer line, was found to express a PDGF-like growth factor. 35S-cysteine labelling and immunoprecipitation revealed the synthesis and secretion of a 31-kDa PDGF-like protein. Serum-free conditioned medium contained PDGF-receptor-competing and mitogenic activity when tested on human fibroblasts. Whereas the receptor-competing activity was fully neutralized by anti-PDGF antibodies, the mitogenic activity was only partially affected. We therefore probed U-1810 mRNA with a panel of growth-factor DNA clones. We found expression of the genes for PDGF A- and B-chains, TGF-alpha, TGF-beta and IGF-II but not EGF or IGF-I. U-1810 cells lacked specific binding sites for PDGF but showed specific binding of EGF and expressed EGF-receptor transcripts. Thus, U 1810 is an example of a human tumor cell line that expresses multiple growth factor genes; in the intact tumor the corresponding growth factors may operate in autocrine stimulation of the tumor cells as well as in paracrine growth reactions (i.e. stroma recruitment). PMID- 3030942 TI - Structure and expression of mos sequences in spontaneous and Moloney murine sarcoma virus-induced yolk sac carcinomas in rats. AB - Facilitation of yolk-sac carcinoma (YSCa) development in fetectomized rats by the Moloney murine sarcoma virus/murine leukemia virus (Mo-MSV/MLV) complex was found to be closely associated with the presence of Mo-MSV sequences in the genomes of the YSCa cells. The virus-induced YSCas consisted of cells of mono- or oligoclonal origin which always contained in their genomes at least I randomly integrated Mo-MSV provirus. In YSCas which developed in the absence of virus, no rearrangement or amplification of c-mos could be detected. In addition, blot hybridization analysis of cellular RNA failed to detect mos-related RNA in cell lines derived from Mo-MSV-induced as well as from non-virally induced YSCas. The methylation level of c-mos DNA was low in all YSCa cell lines. In contrast, v-mos DNA in cell lines derived from Mo-MSV-induced YSCas was heavily methylated. PMID- 3030943 TI - Establishment and characterization of a T-lymphoblastoid cell line MDCC-MTB1 derived from chick lymphocytes infected in vitro with Marek's disease virus serotype 1. AB - Continuously proliferating T-lymphoblastoid cells, named MDCC-MTB1, were obtained by infection of chick embryo lymphocytes with Marek's disease virus serotype I (MDV1) in culture and subsequent cultivation in the presence of human interleukin 2 (IL-2). The MTB1 cells have now been growing well for at least 4 months with a doubling time of about 10 hr, irrespective of the presence of IL-2. The MTB1 cells show lymphoblastoid morphology, and carry T-lymphocyte marker surface antigens and a karyotype of female chick origin with several abnormal chromosomes. Southern blot hybridization showed that they contain about 10 virus genome equivalents/cell of almost the whole MDV genome. Infectious virus could not be rescued from MTB1 cells by co-cultivation of these with chick embryo fibroblasts. In addition, no virus particles were found in thin-sectioned MTB1 cells by electron microscopy. An immunofluorescence test with monoclonal antibody (MAb) to MDV-specific phosphorylated proteins showed that MTB1 cells expressed the MDV-specific antigen in the cytoplasm only after the cells had been treated with 5-iodo-2-deoxyuridine for 48 hr at 41 degrees C. MTB1 cells can form colonies in 0.33% soft agar, and can be transplanted into chicks by i.p. injection. Thus, continuously growing lymphoblastoid cells were obtained by in vitro infection of chick embryo lymphocytes with oncogenic MDV1 and cultivation of the cells in the presence of human IL-2 during the transformation step. These cells appear to show a similar phenotype to an MD lymphoma-derived cell line. PMID- 3030944 TI - In vitro antigenic modulation of human neuroblastoma cells induced by IFN-gamma, retinoic acid and dibutyryl cyclic AMP. AB - Testing with a panel of 26 monoclonal antibodies (MAbs) showed the antigenic profile of 13 human neuroblastoma cell lines to be characterized by a generally poor antigenic expression; therefore, Interferon-gamma (IFN-gamma), dibutyryl cyclic-AMP and retinoic acid were used to analyse the modulation of surface antigenic expression during differentiation. Treatment of neuroblastoma cell lines with IFN-gamma resulted mainly in induction or increase of class-I MHC antigenic expression. Induction of class-II MHC antigens was obtained on only one neuroblastoma cell line out of 13, thus representing an exceptional event. An increase in some other antigens expressed by neuroblastoma cell lines was also observed. In contrast, and in addition to morphological maturation, treatment of these cell lines with the differentiation inducer dibutyryl-cyclic-AMP (dbc-AMP), resulted in general down-modulation of antigenic expression, particularly of neuroblastoma-associated 5A7 or Leu7 antigens. Retinoic acid treatment had no significant effect on MHC antigens, but it decreased expression of 5A7 and Leu7 antigens, and markedly increased the expression of the melanoma-associated antigen Me14-D12. The similarity between the antigenic profile of in vitro differentiated neuroblastoma cells and that of mature ganglioneuroma cells suggests that compounds like cyclic-AMP or retinoic acid are excellent tools for further investigations of the mechanisms of neuroblastoma differentiation and might have important clinical applications. PMID- 3030945 TI - Technetium-99m pyrophosphate single-photon emission computed tomography of the heart in familial amyloid polyneuropathy. AB - A patient with familial amyloid polyneuropathy and congestive heart failure underwent myocardial imaging using technetium-99m pyrophosphate. Planar scintigraphy showed an intense, diffuse biventricular uptake of the radiotracer. Single-photon emission computed tomography demonstrated an unevenly distributed uptake of the isotope. The greatest activity corresponded to regions with marked echocardiographic changes. Emission tomography may aid in assessing the degree and distribution of the infiltrative lesions in cardiac amyloidosis. PMID- 3030946 TI - Trimethoprim-polymyxin B sulfate cream versus fusidic acid cream in the treatment of pyodermas. AB - One hundred patients with a clinical diagnosis of a primary or secondary superficial pyoderma were entered into a double-blind study. They were allocated treatment with either trimethoprim-polymyxin B sulfate cream or fusidic acid cream according to a fully randomized treatment code. Data suitable for evaluation were obtained from 87 patients, and statistical analysis revealed trimethoprim-polymyxin B sulfate (TP) cream to be significantly better than fusidic acid cream in alleviation several of the individual signs and symptoms associated with pyogenic infection of the skin as well as in reducing the overall severity score at the end of the 2-week study period. PMID- 3030947 TI - Bulimic vomiting alters pain tolerance and mood. AB - Bulimia, a disorder of episodic binging and purging, remains without a known etiology. A case report is presented of a patient who attributed bulimic episodes to efforts at inducing euphoria. Experimental pain tolerance was increased by bulimic vomiting, blocked by naloxone, but not by saline. Vomiting was also associated with falls in depression and anxiety. Plasma ACTH and cortisol, putative markers for beta-endorphin, also rose following vomiting. It is hypothesized that in some bulimics, the disorder arises by virtue of an addiction to one's own internally released endogenous opioid peptides. PMID- 3030948 TI - Free radical formation in crystals of guanine hydrochloride dihydrate: an ESR and ENDOR study. AB - Radiation-induced free radical formation in single crystals of guanine hydrochloride dihydrate has been studied at temperatures between 20 and 300 K using ESR and ENDOR spectroscopy. At low temperatures three radical species are trapped. Two of these are the C8 H-addition radical R1 previously analysed by Alexander and Gordy (1967) and the O6-protonated anion radical R2. The third species (R4) remains unidentified. Upon annealing at 280 K for an extended period the protonated anion R2 transforms into a new radical R3 which exhibit a well defined hyperfine pattern but still could not be identified unambiguously. Also radical R4 probably transforms into a new radical (R5) upon such treatment. One proton coupling due to R5 was detected. A scheme of radical reactions incorporating these five radicals is proposed. This scheme also suggests that differences in radical formation between the monohydrate and dihydrate crystals of guanine hydrochloride depends upon differences in the hydrogen bonding network. PMID- 3030949 TI - Radiation cell survival and growth delay studies in multicellular spheroids of small-cell lung carcinoma. AB - The radiation sensitivity of two small-cell lung carcinoma cell lines growing as multicellular spheroids in static culture was determined using clonogenic cell survival and growth delay as endpoints. Growth delay determination suggested that clonogenic cell kill was less than was obtained by direct assay of cell survival. Recovery from potentially lethal damage was assayed in one line (HC12) but was not demonstrable, and clonogenic cell survival decreased with time in treated spheroids with diameters greater than 300 microns which contained a hypoxic cell population. Microscopic examination of the treated spheroids showed the emergence of an abnormal giant-cell population, and the progressive clonogenic cell loss that occurred after treatment was thought to be due to oxygen and nutrient deprivation of the remaining viable cells by this doomed cell population. Correction of the growth delay measurements for changes in cell size and clonogenic cell population allowed correlation of the growth delay and cell survival data. PMID- 3030950 TI - Medical students and community physicians also susceptible to varicella-zoster virus. PMID- 3030951 TI - Nosocomial outbreak of Legionnaires' disease: molecular epidemiology and disease control measures. AB - Molecular laboratory techniques were used to study the epidemiology of an outbreak of nosocomial Legionnaires' disease. All patient isolates were Legionella pneumophila serogroup 1 and showed identical plasmid profiles and reactions with serogroup-specific monoclonal antibodies. L pneumophila was also cultured from four of five cooling tower water samples; however, the isolate from only one tower was serogroup 1 of the same subtype as patient isolates. Since the cases were temporally clustered and epidemiologically associated with exposure to cooling tower aerosols, the single cooling tower implicated by molecular analysis was the most likely source of the outbreak. Chlorination of cooling tower ponds has eradicated the epidemic strain. Since potable water also harbored the infecting organism and was the probable source for cooling tower contamination, decontamination of the hospital water system was also undertaken. Superchlorination of hot water holding tanks to 17 ppm on a weekly basis has effectively eradicated L pneumophila from the potable water system and appears to be a reasonable, simple, and relatively inexpensive alternative to previously described methods of control. PMID- 3030952 TI - Cytomegalovirus. PMID- 3030953 TI - Alpha 1-adrenergic receptor induced subsensitivity and supersensitivity in rabbit iris-ciliary body. Effects on myo-inositol trisphosphate accumulation, arachidonate release, and prostaglandin synthesis. AB - Topical treatment of the rabbit eye with three successive doses of 2% epinephrine resulted in attenuation of the in vitro drug response of the alpha 1-adrenoceptor mediated phosphoinositide hydrolysis system and of alpha 1-adrenergic receptor mediated contraction in the iris. In contrast, sympathetic denervation of the eye resulted in potentiation of these responses. Desensitized tissues showed a significant decrease in epinephrine-induced myo-inositol trisphosphate (IP3) accumulation, 1,2-diacylglycerol (DG) formation, measured as phosphatidate, arachidonic acid (AA) liberation, measured by radiochromatography, prostaglandin (PG) E2 release, measured by radiochromatography and radioimmunoassay, and muscle contraction. Adrenergic desensitization of the eye resulted in attenuation of: The polyphosphoinositide response in the iris, measured both as loss of 32P radioactivity from phosphatidylinositol 4,5-bisphosphate (PIP2) and as IP3 accumulation; the epinephrine-stimulated liberation of AA, from membrane phosphoinositides and other phospholipids, and PGE2 release in the iris; and the epinephrine-induced muscle contraction in the iris dilator. This adrenergic desensitization of the eye is reversible. Surgical sympathetic denervation, previously found to increase alpha 1-adrenoceptor mediated accumulation of IP3 and contraction, increased AA liberation. Dexamethasone blocked the epinephrine induced liberation of AA and PG release, both in vivo and in vitro. These data support the hypothesis that changes in the activity of the alpha 1-adrenergic receptor-mediated phosphoinositide hydrolysis system and its derived second messengers may underlie the mechanism of adrenergic subsensitivity and supersensitivity in the iris-ciliary body. How much the desensitization of alpha 1-adrenergic receptor-mediated responses contribute to the therapeutic action of epinephrine in the eye remains to be determined. PMID- 3030954 TI - Distribution and properties of beta-adrenergic receptors in human iris-ciliary body. AB - beta-Adrenergic receptors from the iris-ciliary body of human eyes removed shortly after death were studied using membranes prepared by isopycnic centrifugation of tissue homogenates. This procedure separates uveal melanin pigment from plasma membranes and reduces nonspecific binding of 125I iodopindolol. The observed binding of 125I-iodopindolol was of high affinity (Kd = 78 +/- 6.6 pM), saturable, and fully reversible (t1/2 = 4.6 min). Scatchard plots were linear and revealed a Bmax of 134 +/- 20 fmol/mg of protein from the whole iris-ciliary body. The affinities of the receptors for a series of agonists and antagonists were determined. The order of potency for the inhibition of the binding of the radioligand by antagonists was ICI 118,551 greater than MK950 greater than propranolol greater than ICI 89,406 greater than metoprolol. This order of potency is characteristic of beta-adrenergic receptors of the beta 2 subtype. Preparations of iris-ciliary body were also subjected to microdissection prior to density gradient centrifugation to permit the study of beta-adrenergic receptors in the ciliary processes, ciliary body, and iris. Each of these regions was found to contain approximately one third of the total number of beta adrenergic receptors in the human iris-ciliary body. The highest density of receptors was located in the ciliary processes (180 +/- 40 fmol/mg of protein), while the density of receptors in the iris (98 +/- 7.5 fmol/mg of protein) and ciliary body (less the processes) (42 +/- 17 fmol/mg of protein) was notably lower. Only beta 2-adrenergic receptors are detectable by competition experiments in the iris-ciliary body as a whole, or in the individual preparations of iris, ciliary processes, or ciliary body; however, microdissection and analysis of beta adrenergic receptor subtypes in isolated ciliary muscle permitted detection of a small number of beta 1-adrenergic receptors. beta 1-Adrenergic receptors comprised about 10% of the total number of beta-adrenergic receptors in the whole iris-ciliary body. The finding that most of the beta-adrenergic receptors in the human iris-ciliary body are of the beta 2 subtype may be of significant therapeutic importance in the medical management of glaucoma. PMID- 3030955 TI - Tear lactoferrin levels in patients with external inflammatory ocular disease. AB - Lactoferrin, an iron complexing protein in normal tears, is an important component of the nonspecific host defense system of the external eye. We measured tear lactoferrin levels in patients with contact lens-induced giant papillary conjunctivitis (GPC) by an enzyme-linked immunosorbent assay (ELISA). Patients with active GPC (N = 26) had significantly reduced tear levels of lactoferrin (0.876 +/- 0.42 mg/ml) compared with normal individuals (N = 12; 1.73 +/- 0.46 mg/ml, P less than 0.0003) and the control contact lens wearers' group (N = 11; 1.57 +/- 0.92 mg/ml, P less than 0.003). Patients with vernal conjunctivitis (N = 10), an ocular disease with similar histopathology, had slightly reduced concentrations of tear lactoferrin (1.22 +/- 0.59 mg/ml). Patients with inactive GPC (N = 7) had normal tear levels of lactoferrin (1.33 +/- 0.49 mg/ml). The lactoferrin to total protein ratio in the tears was significantly reduced in patients with GPC compared to normal subjects, control contact lens wearers, and patients with inactive GPC. The decreased tear levels of lactoferrin in patients with GPC may contribute to increased coating of lenses with bacteria and their products and enhanced ocular inflammation which may play a role in the pathogenesis of GPC. PMID- 3030956 TI - Superiority of antibody versus delayed hypersensitivity in clearance of HSV-1 from eye. AB - The contribution that antibody and delayed type hypersensitivity (DTH) make in promoting HSV-1 clearance from the infected cornea was investigated. Balb/c mice were immunized intravenously or subcutaneously with an attenuated strain of HSV-1 to generate hosts which were antibody-producing DTH-tolerant or antibody producing DTH-responsive. Anti-mu serum treated mice were likewise sensitized intravenously or subcutaneously to obtain hosts which were antibody depressed-DTH tolerant or antibody-depressed DTH-responsive. Eight days after sensitization, these four sensitized groups and unsensitized controls were infected on scarified corneas with a stromal keratitis inducing strain of HSV-1, and the extent of virus replication was determined 1, 3, and 7 days later. Very different results were obtained depending upon the host's immune status. Virus proliferated extensively (greater than 3-4 logs) in the eyes of nonimmune mice and antibody depressed DTH-tolerant hosts during the first 3 days after infection. In striking contrast, HSV-1 could not be detected even 24 hr post challenge in antibody producing DTH-tolerant mice. In fact, such mice cleared virus from the eye as efficiently as immunologically intact hosts. However, in mice with the reverse immune status, ie antibody-depressed DTH-responsive, virus growth was clearly evident (greater than 2-3 logs) during days 1-3, and only thereafter did complete clearance occur. These results indicate that in the sensitized host antibody is both independent of and significantly more effective than DTH in promoting HSV-1 eradication from the infected eye. PMID- 3030957 TI - Two waves of virus following anterior chamber inoculation of HSV-1. AB - Following inoculation of herpes simplex virus type 1 (HSV-1) into the anterior chamber of one eye of a Balb/c mouse, a pattern of ocular retinal disease occurs which is characterized by retinal necrosis, disruption of the retinal architecture of the uninoculated contralateral eye, and sparing of the retina of the virus-inoculated eye. Our direct virus culture studies have revealed that, after uniocular anterior chamber inoculation, virus reaches the uninoculated eye in two temporally separate waves. The first wave of virus is detected in the uninoculated eye as early as one day postinoculation (pi), long before virus is found in either of the optic nerves or the brain. The second wave of virus arrives in this eye between 7 and 10 days pi. Sequentially, the path of the second wave of virus appears to move from the injected eye to (1) the ipsilateral optic nerve, (2) the brain, (3) the contralateral optic nerve, and (4) the posterior segment of the uninoculated contralateral eye, suggesting that interocular spread of the second wave of virus after anterior chamber inoculation occurs via neural pathways. Results of histopathologic examinations and virus culture studies suggest that the early wave of virus contributes to the inflammation observed at the angle structures of the contralateral eye 7-8 days pi and that the second wave of virus accounts for the peak virus titer observed on day 10 pi, a peak which coincides with the destruction of the retina of this eye. It is proposed that the first wave is causally related to the development of retinitis, which occurs as the second wave reaches the retina. PMID- 3030958 TI - Timolol promotes reactivation of latent HSV-1 in the mouse iontophoresis model. AB - The present study examined the effect of topical timolol, a nonspecific beta 1 and beta 2 blocker on reactivation and ocular shedding of latent HSV-1 in an improved mouse iontophoresis model. Latent trigeminal ganglionic infection was established in Balb/C mice following inoculation by corneal scarification with HSV-1 W strain, a clinical isolate, and confirmed by co-cultivation. On day 30, postinfection (pi), the mice were divided into two groups, and treatment begun with coded eye drops (timolol 0.5% or placebo) BID OU for 5 days. On day 31 pi, iontophoresis with 1% 6-hydroxydopamine was performed, and daily treatment with topical epinephrine and 1% prednisolone was administered. Reactivation and recovery of latent HSV-1 was determined by daily ocular swabs, and characteristic HSV-1 cytopathic effect in Vero cells. Results demonstrated that the timolol treated group had a significantly greater number of positive eyes, multiple shedding episodes, and total shedding days compared to the control group. We conclude that beta blockade promotes recurrent ocular shedding induced by epinephrine in the mouse iontophoresis latency model. PMID- 3030959 TI - Timolol induces HSV-1 ocular shedding in the latently infected rabbit. AB - Timolol iontophoresis into the eye can induce herpes simplex virus type 1 (HSV-1) shedding in rabbits latently infected with HSV-1 strain McKrae. Anodal iontophoresis of 0.01% timolol was done at 0.8 mAmp for 8 min once a day for 3 consecutive days. Viral shedding was determined by the presence of HSV-1 in the preocular tear film obtained by eye swabs. In two experiments, iontophoresis of 0.01% timolol resulted in all eyes (18/18) shedding HSV-1 for an average duration of 4.3 days. When 5.0% timolol was applied topically to rabbit eyes supersensitized by iontophoresis of 6-hydroxydopamine (6-HD), all eyes (10/10) shed virus for an average duration of 2.9 days. All eyes (12/12) receiving iontophoresis of 6-HD, pre- and posttreatment with topical application of 5.0% timolol, and posttreatment with topical application of 1.0% epinephrine shed virus for an average duration of 3.6 days. Eyes treated with topical application of 5.0% timolol alone showed no difference in HSV-1 ocular shedding, compared with untreated eyes. We concluded that both iontophoresis of 0.01% timolol and topical application of 5.0% timolol to adrenergically supersensitized eyes induced HSV-1 shedding reliably and with a high frequency, and that topically applied timolol does not block the HSV-1 ocular shedding induced by epinephrine in adrenergically supersensitized eyes. PMID- 3030960 TI - Inhibition of herpes simplex virus replication in the mouse cornea by drug containing immunoliposomes. AB - Monoclonal antibody to HSV glycoprotein D was derivatized with palmitic acid and incorporated into liposomes. These immunoliposomes bound specifically to intact mouse corneas infected with HSV-1 in vitro. Furthermore, in yield reduction assays, anti-gD immunoliposomes loaded with acyclovir proved far more effective at inhibiting viral replication in the cornea than free drug or drug delivered in untargeted liposomes. Site-specific sustained release immunoliposomes of this type are potentially an improved vehicle for drug delivery in the treatment of ocular HSV. PMID- 3030961 TI - Theophylline abolishes the light peak in perfused cat eyes. AB - Theophylline, a phosphodiesterase inhibitor, was applied to the arterially perfused cat eye in vitro. At a concentration of 850 microM, theophylline abolished the light peak of the DC-ERG and led to a 20% increase in perfusate flow to the eye. The effects were reversible. An experimentally-induced increase in flow per se by 20% resulted in a slight enhancement of the light peak. The data suggest the involvement of cyclic nucleotides in the generation of the light peak. Theophylline also caused a decrease in the standing potential of the eye, an increase in the b-wave amplitude, and a decrease in the c-wave amplitude in a dose-dependent and reversible manner. PMID- 3030962 TI - Contrast enhancement of the liver. Evaluation by automated contiguous pixel search. AB - Eight dogs were infused with one of three different contrast agents: meglumine diatrizoate, iosulamide meglumine, or an ethiodized oil emulsion (EOE 13). Image intensity in the liver was evaluated globally and regionally by means of an automated pixel sampling method to determine differences in the rate of enhancement produced by the three agents. The results of the automated method were compared with those of the standard manual cursor method. The automated method showed that liver parenchyma was enhanced uniformly by all three contrast agents. The maximum degree of enhancement (Mean +/- SEM) for the three agents was diatrizoate, 12.0 +/- 3.5 Hounsfield units (HU); iosluamide, 21.3 +/- 3.5 HU; and EOE 13, 37.2 +/- 4.25 HU. With the manual cursor method, contrast enhancement was about 20% more than estimated by the automated method. The automated method is better for evaluating the magnitude and pattern of contrast agent enhancement of the entire liver, since the currently employed cursor technique requires multiple evaluations to evaluate the entire liver. PMID- 3030963 TI - Comparison of two 99mTc(V)-dimercaptosuccinic acid preparations. AB - 99mTc(V)-DMSA labelled either by increasing the pH together with a low concentration of Sn, or by increasing the pH only are compared in vitro and in vivo. The gel chromatographic separation shows that after incubation with blood, all the 99mTc(V)-DMSA preparations are stable and do not bind to the plasma proteins. The behavior in rats as well as in clinical studies does not demonstrate any distinctive characteristics between the different radiomolecules. It seems that the pH of the solution is an important factor which controls the formation of 99mTc(V)-DMSA. PMID- 3030964 TI - Zirconium molybdate gel as a generator for technetium-99m--I. The concept and its evaluation. AB - A new 99mTc generator for the preparation of radiopharmaceuticals has been developed. Its elution characteristics and chemical stability merit its consideration as a replacement for present generators. The 99Mo is present as a zirconium molybdate gel, the high molybdate content of which allows the use of (n, gamma) 99Mo. The resulting generator is as effective and as convenient to use as chromatographic generators that are based on fission product 99Mo. Laboratory evaluations of the concept at low levels of radioactivity described in this paper show that the gel can be prepared in a form which is stable when used in a generator and from which TcO4- can be eluted in yields of 80-85%. The influence of formulation variables on gel properties was established. The performance of this generator using 99Mo with specific activities between 4 and 13 GBq g-1 is described in a further paper. PMID- 3030965 TI - Zirconium molybdate gel as a generator for technetium-99m--II. High activity generators. AB - The characteristics of zirconium molybdate gels were described in Part I of this paper. Here it is shown that zirconium molybdate gel, prepared from neutron irradiated molybdenum oxide with 99Mo specific activities between 4.3 and 13.5 GBq g-1 is sufficiently stable for use as a matrix in preparation of high activity 99mTc generators. Elution efficiencies of generators containing from 2 to 6 g of gel ranged from 83 to 50% for 10 mL elutions. Generators containing 150 g of gel were eluted with efficiencies of 80-98%, with the activity peak in the first 50 mL. Radionuclidic impurities were considerably below the limits set by the British Pharmacopoeia. Although the pH of eluates was less than 4, subsequent treatment with a small zirconium oxide bed provided adjustment to within BP limits, and reduced 99Mo impurity levels tenfold to values near 10(-4)%. No significant differences were found between the biodistributions in rats of radiopharmaceuticals prepared with pertechnetate from gel generators and those from other generators. PMID- 3030966 TI - The standardization of samarium-153. AB - Samarium-153 has been standardized by 4 pi beta liquid-scintillation counting, with an uncertainty of 0.4%. The probability per decay for the 103.2-keV gamma ray was measured, using two germanium detectors, to be 0.298 +/- 0.004. The half life, based on liquid-scintillation measurements over 6.4 half lives and pressurized-ionization-chamber measurements over four half lives, was found to be 46.27 +/- 0.02 h. The uncertainties given are one estimated standard deviation (of the mean when applicable) for random and non-random components. PMID- 3030967 TI - Chemical and biological properties of 2,6-diisopropyl IDA labelled with 99mTc. AB - A procedure is described for the synthesis of 2,6-diisopropyl IDA (DISIDA) and for preparation of the radiopharmaceutical by "instant technique". The chemical parameters affecting the formation of the Sn(II)-DISIDA complex were also determined. Biological studies involved lyophylic measurements, determination of binding constants as well as rate of binding to proteins and biodistribution. Clinical investigation demonstrated good hepatobiliary features of this radiopharmaceutical. PMID- 3030968 TI - Fast chemical synthesis of [75Se]L-selenomethionine. AB - A fast chemical synthesis of high specific activity [75Se]L-selenomethionine (10 Ci/mmol--370 GBq/mmol) is described with a view to 73Se labeling and PET studies. The overall radiochemical yield of the preparation is better than 80%. The purification method uses commercially available reverse phase HPLC columns and 9% NaCl as mobile phase. The final labeled compound is obtained in less than 3 h and the chemical, radiochemical and optical purities of the L-isomer are higher than 99.0%. PMID- 3030969 TI - Preparation of [11C]buprenorphine--a potential radioligand for the study of the opiate receptor system in vivo. AB - A method is described for the preparation of [11C]buprenorphine in high specific activity, based on the reaction of N-(de-cyclopropylmethyl)buprenorphine with "no carrier added" [1-11C]cyclopropanecarbonyl chloride followed by reduction with lithium aluminium hydride. The [1-11C]cyclopropanecarbonyl chloride is itself prepared from cyclotron-produced [11C]carbon dioxide. The overall preparation time is 57 min from the end of radionuclide production, and the radiochemical yield is ca 20%, (decay-corrected from [11C]-carbon dioxide). [11C]Buprenorphine has potential as a radioligand for the study of the opiate receptor system in vivo by means of position emission tomography. PMID- 3030970 TI - [Separation of [Rh-103m]-rhodocene derivatives from the parent [103Ru]ruthenocene derivatives and their organ distribution]. AB - The radioactive decay of [103Ru]ruthenocene derivatives leads to 103mRh labelled rhodocinium derivatives, which can be separated by the extraction of a lipophilic solution of the ruthenocen derivate with water. The separation factor 103mRh/103Ru reaches values of 32:1 Rh3+ ions are not liberated and extracted. The organ distribution of the 103mRh labelled rhodocinium derivatives gained from ruthenocene and from N-isopropyl-ruthenocene amphetamine is different from the distribution of the parent ruthenocene compound. The liver and kidney uptake of the rhodocinium-amphetamine is much higher than the uptake with ruthenocene amphetamine. PMID- 3030971 TI - Conversion of 111In-oxine to 111In trichloride by acid hydrolysis. AB - The lipophilic complex 111In-oxine was converted back to 111InCl3 by acid hydrolysis with 0.1 N HCl. Greater than 99% of the starting material was converted to 111InCl3 with a purity of 99.6%. Unlabeled oxine was completely removed by chloroform extraction. Current experimental data suggest that 111In oxine is unstable in weak acidic medium. PMID- 3030972 TI - A note on the effect of gamma-rays on cefamandole and oxacillin. AB - The feasibility of the radiation sterilization of two beta-lactam antibiotic powders, cefamandole nafate and oxacillin sodium, has been examined by subjecting them to a range gamma-radiation doses, followed by chemical and microbiological analyses. It would appear feasible to radiation sterilize oxacillin sodium. The radiation sterilization of cefamandole nafate may be realistic at low doses or under conditions that minimize radiolysis. PMID- 3030974 TI - Incidence of rotavirus infection in pediatric outpatients with gastroenteritis. PMID- 3030973 TI - [Ultrastructure of the vestibular apparatus in patients with Meniere's disease]. AB - The vestibular labyrinth of eight patients with Meniere's disease obtained by translabyrinthine vestibular neurectomy or transtympanal labyrinthectomy was studied with the light microscope and transmission electron microscope. Previous invasive therapy was transtympanal application of gentamicin in one patient and endolymphatic sac surgery in two patients. Some of the pathological findings were unspecific, e.g. mucoid degeneration of the subepithelial spaces, fibrous long spacing collagen and high amount of lipofuscin in different cell types. Cuticular laminated structures were seen in some of the specimen. A most interesting finding was the discrepancy of large numbers of sub- and intraepithelial nerve fibers and a significant degeneration of vestibular sensory cells with vacuoles and filament bundles. This points out the importance of combining labyrinthectomy and vestibular neurectomy in late stages of unilateral Meniere's disease. PMID- 3030975 TI - Practical aspects of angiotensin-converting enzyme inhibition. Proceedings of a symposium. July 25 and 26, 1986, Denver, Colorado. PMID- 3030976 TI - Introduction to the symposium on practical aspects of angiotensin-converting enzyme inhibition. PMID- 3030977 TI - The antihypertensive effect of angiotensin-converting enzyme inhibition: a multifaceted mechanism of action. PMID- 3030978 TI - An overview of angiotensin-converting enzyme inhibitors in the treatment of hypertension. PMID- 3030979 TI - Metabolic consequences of antihypertensive therapy. PMID- 3030980 TI - Tailoring antihypertensive treatment: which patients, what therapy? PMID- 3030981 TI - Angiotensin-converting enzyme inhibition therapy in heart failure: practical guidelines. PMID- 3030982 TI - Transplacental uptake of WR 2721 by the rat embryo. AB - On day 14 post-conception, near the end of the period of major organogenesis, pregnant rats were injected intravenously or intraperitoneally with WR 2721 spiked with 14C-WR 2721. The radioprotectant was shown to cross the placenta rapidly when administered by either route, and the concentration of WR 2721 in the embryos, placentae, and maternal blood plasma was determined during the period 5 to 90 minutes following administration. The concentration of WR 2721 increased continuously in the embryos during this period and did so against a decreasing concentration in the maternal blood. Injection of WR 2721 at 100 mg/kg of maternal body weight resulted in the presence of 8-9-mg/kg embryo weight; this embryo level is about 1/2 the injected dose of WR 2721 currently being used in human radiotherapy trials, that is, 20 mg/kg (740 mg/m2) body weight. Previous toxicity studies of 9, 11, and 14 day rat embryos have shown that this 100 mg/kg dose is much below the level which produces embryotoxic effects. PMID- 3030983 TI - How to use EEG/ERBP phenomena. AB - Hitherto the study of cerebral slow waves has been mainly empirical phenomenology, in spite of 50 years of effort to discover their origins and possible functions. Since 1970 the increasing use of both analog and digital filters, combined with more sensitive averaging techniques, has led to better understanding of some possible functional significance for the electroencephalogram (EEG) and the endogenous slow waves of event-related brain potentials (ERBP). The developing concepts and mathematics of non-linear dynamicsand coupled oscillators, when considered in the light of cerebral circuit geometry and topology, of local neuronal circuitry, and of both observed and experienced biological behavior, now offer hope for future, more complete, understanding of the development of 'conscious and goal-directed' activity from robotic and reflexive stimulus-response paradigms. Immediate practical results of a biophysical approach to brain waves should be: Models of vertebrate brains as distributed systems of non-linearly coupled oscillators; and revision of traditional methodologies for recording and interpreting EEG/ERBP data. PMID- 3030984 TI - Clinical features and treatment of renal tubular acidosis in two horses. AB - Two horses were admitted separately for evaluation and treatment of profound hyperchloremic metabolic acidosis without azotemia. One, an 11-year-old Quarter Horse mare, had been depressed and ataxic for 2 days. The other, a 2-year-old Quarter Horse colt, had a 6-week history of depression, anorexia, and weight loss. Both horses responded to fluid and electrolyte therapy, but required daily oral administration of sodium bicarbonate for maintenance. In each case, the diagnosis was renal tubular acidosis. PMID- 3030985 TI - Human papillomavirus type 16 transformation of rat 3Y1 cells. AB - The molecularly cloned, prototype human papillomavirus (HPV) 16 DNA has a single base deletion in the El gene. We reconstructed a putative non-defective HPV 16 genome with an uninterrupted El gene, and examined its biological functions. The reconstructed HPV 16 DNA formed foci of morphologically transformed cells in transfected rat 3Y1 cell cultures (an immortalized normal cell line). The transformed rat cells contained transcriptionally active HPV 16 DNA, which appeared to be integrated within the cell DNA, and formed colonies in soft agar. PMID- 3030986 TI - The inhibition of neoplastic cell proliferation with human natural tumor necrosis factor. AB - Purified human natural tumor necrosis factor (n-TNF) was prepared by stimulating human leukemic B cell line (BALL-1) with Sendai virus. The colony formations of all of 18 human cancer-derived abnormal cell lines were suppressed by 10(1)-10(6) U/ml of n-TNF, while n-TNF was nontoxic to all human normal fibroblast cells. This in vitro inhibition of cell growth was reversible. In breast adenocarcinoma MCF7 cells treated with n-TNF a specific decrease of DNA synthesis was observed, and DNA histograms showed a block at G1 in the cell cycle. In vivo studies revealed that n-TNF suppressed the tumor growth of murine Meth A sarcoma, human renal adenocarcinoma (ACHN), malignant melanoma (SK-MEL-28) and glioblastoma (U 373 MG). Isobologram analysis showed that n-TNF synergistically inhibited cell growth in combination with human natural interferon (IFN)-a. In vivo synergism of n-TNF and IFN-a was also found in the U-373 MG tumor model implanted into nude mice. PMID- 3030988 TI - 1 alpha,25-Dihydroxyvitamin D3 induces anchorage-independent growth and c-Ki-ras expression of BALB/3T3 and NIH/3T3 cells. AB - 1 alpha,25-Dihydroxyvitamin D3, a hormonally active form of vitamin D, induces anchorage-independent growth of BALB/3T3 A31-1-1 and NIH/3T3 cells with concomitant increase of their mRNA level of c-Ki-ras but not of c-Ha-ras or c myc, through a receptor-mediated mechanism. Under the same conditions, 12-O tetradecanoylphorbol-13-acetate did not induce anchorage-independent growth in these cell lines. PMID- 3030987 TI - The bovine leukemia virus X region encodes a trans-activator of its long terminal repeat. AB - We constructed a fusion plasmid, pMX-I, by which the major open reading frame, X I, of the bovine leukemia virus (BLV) X gene was expressed under control of the mouse metallothionein promoter. pMX-I was cotransfected into CV1 monkey kidney cells together with another construct containing the BLV long terminal repeat (LTR) linked to the chloramphenicol acetyltransferase (CAT) structural gene. The result of assay of CAT synthesis suggests that the X-I product functions as a trans-acting activation factor of the BLV LTR. PMID- 3030989 TI - Design of LHRH agonist drug candidates. PMID- 3030990 TI - Characterization of beta-adrenergic receptors in dispersed rat testicular interstitial cells. AB - Recent studies have shown that beta-adrenergic agents stimulate steroidogenesis and cyclic AMP formation in mouse Leydig cells in culture. To obtain information about the possible presence and the characteristics of a beta-adrenergic receptor in rat testicular interstitial cells, the potent beta-adrenergic antagonist [125I]cyanopindolol (CYP) was used as ligand. Interstitial cells prepared by collagenase dispersion from rat testis were incubated with the ligand for 2 h at room temperature. [125I]cyanopindolol binds to a single class of high affinity sites at an apparent KD value of 15 pM. A number of sites of 6,600 sites/cell is measured when 0.1 microM (-) propranolol is used to determine non-specific binding. The order of potency of a series of agonists competing for [125I]cyanopindolol binding is consistent with the interaction of a beta 2 subtype receptor: zinterol greater than (-) isoproterenol greater than (-) epinephrine = salbutamol much greater than (-) norepinephrine. In addition, it was observed that the potency of a large series of specific beta 1 and beta 2 synthetic compounds for displacing [125I]cyanopindolol in rat interstitial cells is similar to the potency observed for these compounds in a typical beta 2 adrenergic tissue, the rat lung. For example, the potency of zinterol, a specific beta 2-adrenergic agonist, is 10 times higher in interstitial cells and lung than in rat heart, a typical beta 1-adrenergic tissue. Inversely, practolol, a typical beta 1-antagonist, is about 50 times more potent in rat heart than in interstitial cells and lung.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3030991 TI - 6-(E)-acetylmethylenepenicillanic acid, a potent beta-lactamase inhibitor. PMID- 3030992 TI - Inheritance and interaction of immune traits in beef calves. AB - Three sets of blood samples were obtained from beef calves of two experimental populations and assayed for various immunological measurements. The first set of samples was taken between 24 and 48 h after birth and quantified for IgG1 concentration. A second set was taken immediately prior to vaccination for infectious bovine rhinotracheitis virus (IBRV; at an average age of 164 d) and a third set taken 60 d post-vaccination. These later samples were quantified for antibodies specific to IBRV. Level of complement C3 was also quantified in the samples taken immediately prior to vaccination. Three hundred sixty-seven calves were from four Hereford lines; three lines were previously selected for growth traits and the fourth was a randomly selected control line. There were no consistent differences in immune traits among these lines. The second group of 165 animals were Angus, Hereford and Red Poll calves. While Angus calves had a higher mean IgG1 concentration at 24 to 48 h of age than Hereford or Red Poll calves, no differences among breeds were found for the other immune traits measured. Calves from older dams (greater than 3 yr old) tended to have higher mean IgG1 concentrations, pre-vaccination IBRV antibody titers and complement C3 levels than calves from 2- and 3-yr-old cows. However, these calves had lower 60 d post-vaccination IBRV titers than calves from the younger cows (P less than .05). As pre-vaccination IBRV antibody titer increased, post-vaccination IBRV antibody titer decreased (P less than .05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3030993 TI - Effect of dietary electrolyte balance on growth and acid-base status in swine. AB - The effect of dietary electrolyte balance on pigs fed lysine- or tryptophan adequate or tryptophan-deficient diets was investigated in four experiments using 8- to 12-wk-old pigs. Electrolyte balance, expressed as Na+K-Cl in meq/kg of diet, was varied by altering dietary levels of Na and Cl while holding all other minerals constant. In two experiments in which the basal diet contained a balance of 135 meq/kg, simple lysine or tryptophan deficiences caused depressed growth, feed intake and efficiency of feed utilization, but none of these responses was altered by dietary supplementation with NaHCO3. In one experiment in which the electrolyte balance of the basal diet was 61 meq/kg and in which both lysine and tryptophan were limiting. NaHCO3 supplementation significantly increased growth and feed intake. This did not occur if the diet was also supplemented with tryptophan. A final experiment was conducted to determine the response of pigs to a range of electrolyte balance (-85 to 341 meq/kg) in a practical corn-soy diet containing adequate levels of all amino acids. Growth and feed intake appeared to be maximal for balances of 0 to 341 meq/kg Na+K-Cl, but were decreased at -85 meq/kg (P less than .05). Acid-base balance was adversely affected at 0 meq/kg. The results suggest that the response of lysine-deficient pigs to sodium bicarbonate is dependent upon the electrolyte balance of the diet, and also is influenced by other dietary amino acids. PMID- 3030994 TI - Serum isocitrate dehydrogenase during polybrominated biphenyl toxicosis in dairy cattle. AB - Bovine serum isocitrate dehydrogenase (sICDH) was investigated in dairy cattle as a clinical measurement indicative of hepatic injury. Conditions for optimization of isocitrate dehydrogenase assays for bovine serum are described. Assays of sICDH in normal cattle show average activities of .814 (SD = .202) units/ml serum with a range of .316 to 1.268 for 83 samples taken from 32 animals. Investigation of sICDH in pregnant dairy cattle experimentally dosed with polybrominated biphenyl (PBB) showed no discernible elevations until doses were sufficient to cause toxicosis (25,000 mg PBB/d). Cows lethally dosed with 25,000 mg PBB/d had moderate elevations of sICDH (approximately a twofold increase) concomitant with severe toxicosis in some but not all animals. This PBB dose also caused abortion or fetal death in pregnant animals; elevation of sICDH in these animals was coincident with fetal trauma. This suggests that sICDH may be influenced by fetoplacental contributions in pregnant animals. Non-pregnant cows, intoxicated with PBB, had minimal sICDH elevation as compared with 10-fold in a calf with experimentally induced hepatotoxicity (thioacetamide). This observation was consistent with histopathological findings of minimal, if any, hepatic involvement in dairy cattle lethally intoxicated with PBB. Serum isocitrate dehydrogenase appears to be a useful adjunct to the ordinary complement of serum chemistries used for clinical diagnosis; however, it does not appear to reflect exclusively hepatic injury. PMID- 3030995 TI - Comparison of induced corpora lutea from prepuberal gilts and spontaneous corpora lutea from mature gilts: in vitro progesterone production. AB - Prepuberal (P) gilts were induced to ovulate with pregnant mare serum gonadotropin followed 72 h later by human chorionic gonadotropin (hCG). Three P gilts and three mature (M) gilts each were ovariectomized on d 10, 14, 18, 22 and 26 (d 0 = day of hCG for P gilts and onset of estrus for M gilts). Gilts ovariectomized on d 14, 18, 22 and 26 were hysterectomized on d 6 to ensure maintenance of the corpora lutea (CL). Two to five grams of minced luteal tissue were dispersed using collagenase and hyaluronidase in HEPES buffered salt solution supplemented with glucose and bovine serum albumin. Dispersed cells were rinsed in Dulbecco's Modified Eagle Medium (DMEM), counted (ratio of large to total number of luteal cells determined) and then incubated for 1 h in DMEM. With aliquots standardized to 2.5 X 10(4) viable, large cells (greater than 25 micron diameter) were incubated in 1 ml DMEM for 2 h in the presence of either 10, 50, 100 or 1,000 ng luteinizing hormone (LH); .1, 1, 10 or 100 ng hCG; 10, 100 or 1,000 ng norepinephrine (NE) or either .75, or 1.5 mM dibutyrl cyclic adenosine monophosphate (dbcAMP). Progesterone (P4) in the medium was quantified by radioimmunoassay. Basal P4 production (no P4 stimulator added to the medium) on d 10, 14, 18, 22 and 26 for P gilts was 246 +/- 9, 66 +/- 4, 64 +/- 6, 41 +/- 3 and 69 +/- 6 ng/ml medium, respectively, and for M gilts was 281 +/- 12, 128 +/- 8, 53 +/- 4, 82 +/- 6, 101 +/- 5 ng/ml medium, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3030997 TI - Molecular relationships between trimethoprim R plasmids obtained from human and animal sources. AB - A total of 65 trimethoprim R plasmids (35 obtained from human strains and 30 from animal strains of Escherichia coli) were examined by the technique of restriction endonuclease fingerprinting. Plasmids belonging to incompatibility groups B, FII and I delta obtained from human and animal sources showed close similarities within each group. Plasmids belonging to incompatibility groups I alpha and P were also obtained from both human and animal sources, but there was no conclusive evidence of close relationships within the groups. Restriction endonuclease fingerprinting was found to be useful for obtaining information about the epidemiology of R plasmids. Its main limitation in this study related to broad host range plasmids of the P incompatibility group, some of which contained very few sites susceptible to cleavage by the restriction endonucleases tested. PMID- 3030996 TI - Characterization of in situ nitrogen and fiber digestion and bacterial nitrogen contamination of hay crop forages preserved at different dry matter percentages. AB - Alfalfa, red clover, orchardgrass and timothy were harvested in the vegetative stage, wilted and stored as hay, or ensiled in small batch silos (20 kg) at 60, 40 or 20% (direct cut) dry matter and were analyzed for compositional differences. A ruminally cannulated lactating cow, consuming 50% of her dry matter intake from hay crop silage, was used to measure in situ dry matter, N, neutral detergent fiber and acid detergent fiber disappearance. Diaminopimelic acid was used as a bacterial marker to correct for bacterial N contamination for in situ residual N. Fibrous components tended to become concentrated as percent dry matter at preservation decreased, presumably associated with leaching of water solubles during storage. For most forages, as dry matter percentage of preservation decreased, water soluble dry matter and N increased, with a concomitant increase of ruminally nondigested dry matter. Specific trends in coefficients of digestion associated with forage type or preservation dry matter percentage were not observed for dry matter, N, neutral detergent fiber or acid detergent fiber. Correction for contamination by bacterial N decreased lag time in digestion and altered rates of N digestion compared with noncorrected rates. Linear and quadratic bacterial N contamination profiles were observed with time of ruminal incubation. Rate of digestion of N was highly correlated with fibrous component concentration, and to a lesser extent to rate of neutral and acid detergent fiber digestion. Dry matter percentage at preservation had a variable effect on ruminal digestion rate of dry matter and N, which varied with forage type and had no effect on neutral detergent and acid detergent fiber digestion rates. Correction for bacterial N contamination should be considered when establishing N digestion rates for forage by the in situ technique. PMID- 3030998 TI - [Clinical study of sensory disorders of the hand in leprosy]. PMID- 3030999 TI - Comparison of four beta-lactamase inhibitors in combination with ampicillin against Mycobacterium tuberculosis. AB - The in-vitro activity of a beta-lactamase inhibitor (clavulanic acid, sulbactam, BL-P2013 or BL-P2090) in combination with ampicillin against 13 isolates of Mycobacterium tuberculosis was determined by broth dilution. The agents were tested in 1:1 molar ratio of ampicillin to beta-lactamase inhibitor. The beta lactamase inhibitors alone did not inhibit growth at the highest concentration tested. The MIC90 for ampicillin alone was greater than 32 mg/l. Clavulanic acid plus ampicillin was the most active combination with an MIC90 of 11 microM (4 mg/l of ampicillin). The MIC90 of ampicillin in combination with sulbactam, BL P2013 or BL-P2090 was 23 microM (8 mg/l of ampicillin). A 1:1 ratio of ampicillin to inhibitor was more active than was a 2:1 ratio. The addition of a beta lactamase inhibitor to ampicillin greatly improves its in-vitro activity against M. tuberculosis. PMID- 3031001 TI - cAMP suppresses CA2+-dependent electrical activity of airway smooth muscle induced by TEA. AB - Using intracellular microelectrodes, we investigated whether exogenous dibutyryl adenosine 3',5'-cyclic monophosphate (DBcAMP) or forskolin influenced the electrical effects of tetraethylammonium (TEA) on canine tracheal smooth muscle. We found that 20 mM TEA depolarized airway smooth muscle cells from a resting membrane potential (Em) of -59 +/- 4 mV (mean +/- SD) to -45 +/- 2 mV and caused spontaneous action potentials (AP's) to develop, which were 33 +/- 2 mV in amplitude. These were totally abolished in 0 Ca2+ solution. DBcAMP (1 mM) suppressed the development of this TEA-induced electrical activity and the phasic contractions electrically coupled to it. DBcAMP had no significant effect on Em in the absence of TEA however. Forskolin (1 microM) produced similar effects. Our findings suggest that Ca2+ is the principal ion responsible for the inward current associated with the TEA-induced AP's in airway smooth muscle, and that adenosine 3',5'-cyclic monophosphate may suppress the electrogenesis of this current. PMID- 3031000 TI - Interleukin-1-induced granulocytopenia and pulmonary leukostasis in rabbits. AB - Pulmonary leukostasis is a postulated prerequisite lesion for acute lung injury. Interleukin-1 (IL-1) mediates components of the acute-phase response, stimulates granulocyte metabolism and secretion, and augments endothelial adhesiveness. We studied the effects of murine IL-1 infusion on circulating granulocytes, their sequestration within the pulmonary microvasculature, lung water, and bronchoalveolar lavage fluid (BALF) protein concentration in rabbits at 3 and 24 h after infusion. IL-1 administration induced significant (P less than 0.01) granulocytopenia compared with saline-injected controls and at 3 h induced significant increases in both mean alveolar septal wall granulocytes per high power field (HPF) (P less than 0.001) and mean myeloperoxidase (MPO) activity per gram lung tissue (P less than 0.001). At 24 h, IL-1 induced a marked granulocytosis and again significantly increased both mean alveolar septal wall granulocytes per HPF (P less than 0.001) and lung MPO (P less than 0.01). Increased lung water or BALF protein concentration could not be demonstrated in animals killed at either 3 or 24 h after IL-1 administration. Therefore, IL-1 can induce an early profound granulocytopenia followed by later granulocytosis, as well as sustained pulmonary leukostasis in the absence of detectable pulmonary edema formation or an alveolar-capillary leak. PMID- 3031003 TI - Neutrophil collagenase in rheumatoid interstitial lung disease. AB - Ten patients with rheumatoid arthritis were evaluated by bronchoalveolar lavage. Five patients (group I) had interstitial lung disease by physiological and radiographic criteria, whereas five (group II) had no evidence of lung disease. Lavage fluid from four of the five group I patients contained an active collagenase which by inhibitory profile and substrate specificity appeared to be of neutrophil origin. None of the group II patients demonstrated lavage fluid collagenase. Treatment of lavage fluid with trypsin failed to uncover latent collagenase activity in either group, suggesting that the collagenase is present entirely in an active form. These findings parallel those observed in idiopathic pulmonary fibrosis and suggest a potential pathogenetic role for collagenase in rheumatoid interstitial lung disease. PMID- 3031002 TI - Pharmacological modification of pulmonary vascular injury: possible role of cAMP. AB - Experiments were designed to test the hypothesis that drugs which increase adenosine 3',5'-cyclic monophosphate (cAMP) in the lung would prevent the pulmonary hypertension and the increase in vascular permeability caused by the infusion of the oxidant lipid peroxide, tert-butyl hydroperoxide (t-bu-OOH), in isolated rabbit lungs perfused with Krebs-Henseleit buffer. Pretreatment with indomethacin or verapamil was also studied, since these drugs block the increase in pulmonary arterial pressure caused by t-bu-OOH. Indomethacin or verapamil prevented the pulmonary hypertension but did not prevent the increase in permeability caused by t-bu-OOH. Consequently, indomethacin or verapamil treatment partially reduced the gain in lung weight caused by t-bu-OOH. In contrast, pretreatment with isoproterenol, prostaglandin E1, or a cAMP analogue not only prevented the pulmonary hypertension but also inhibited the increase in vascular permeability caused by t-bu-OOH. Consequently, these drugs completely blocked the gain in lung weight caused by t-bu-OOH. Posttreatment with aminophylline or the cAMP analogue also significantly reduced the gain in lung weight caused by t-bu-OOH. These results indicate that pharmacological therapy can reduce the pulmonary hypertension and the increase in vascular permeability caused by the infusion of a lipid hydroperoxide. Since isoproterenol, aminophylline, prostaglandin E1, and a cAMP analogue all had similar effects, the results suggest that the likely common mechanism for their protective effect is an increase in cAMP. PMID- 3031004 TI - Prejunctional beta 1-adrenoceptors inhibit cholinergic transmission in canine bronchi. AB - The aim of the present study was to determine in canine bronchi the effects produced by norepinephrine (released from adrenergic nerve terminals) on cholinergic neurotransmission. Electrical stimulation of canine bronchi activates cholinergic and adrenergic nerve fibers. The adrenergic neuronal blocker, bretylium tosylate, inhibited the increase in [3H]norepinephrine overflow evoked by electrical stimulation but did not prevent that caused by the indirect sympathomimetic tyramine. During blockade of the exocytotic release of norepinephrine with bretylium, the pharmacological displacement of the sympathetic neurotransmitter by tyramine significantly decreased the contractions evoked by electrical stimulation but did not affect contractions caused by exogenous acetylcholine. Metoprolol, a beta 1-adrenergic antagonist, abolished and propranolol significantly reduced the effect of tyramine during electrical stimulation. alpha 2-Adrenergic blockade, beta 2-adrenergic blockade, or removal of the epithelium did not significantly affect the response to tyramine. These results suggest that norepinephrine when released from sympathetic nerve endings can activate prejunctional inhibitory beta 1-adrenoceptors to depress cholinergic neurotransmission in the bronchial wall. PMID- 3031005 TI - Use of solid phase Florisil cartridges to separate fat from semivolatile organic compounds in adipose tissue. AB - A quick method for separation of semivolatile organic compounds from fat in adipose tissue has been developed. This method uses commercially available solid phase cartridges for sample cleanup. The results indicate that the recoveries, from hexane-extracted fat, of 4 representative classes of organic compounds range from 86.2 to 116%. The solid phase cartridges provide excellent separations of the fat from the analytes; no extraneous interference peaks were detected in the gas chromatograms. The method requires only 0.1 g sample and is quick and simple to use. Although results are reported for samples containing 1-14 ppm, the final extract can be concentrated to a volume allowing detection between 10 and 100 ppb. PMID- 3031006 TI - Sarcoidosis and ACE assay. PMID- 3031007 TI - Diagnostic value of SACE. PMID- 3031008 TI - Toxic accumulation of alpha-ketobutyrate caused by inhibition of the branched chain amino acid biosynthetic enzyme acetolactate synthase in Salmonella typhimurium. AB - Biochemical and genetic analyses of the bacterium Salmonella typhimurium suggest that accumulation of alpha-ketobutyrate partially mediates the herbicidal activity of acetolactate synthase inhibitors. Growth inhibition of wild-type bacteria by the herbicide sulfometuron methyl was prevented by supplementing the medium with isoleucine, an allosteric inhibitor of threonine deaminase-catalyzed synthesis of alpha-ketobutyrate. In contrast, isoleucine did not rescue the growth of a mutant containing a threonine deaminase unresponsive to isoleucine. Moreover, the hypersensitivity of seven Tn10 insertion mutants to growth inhibition by sulfometuron methyl and alpha-ketobutyrate correlated with their inability to convert alpha-ketobutyrate to less noxious metabolites. We propose that alpha-ketobutyrate accumulation is an important component of sulfonylurea and imidazolinone herbicide action. PMID- 3031009 TI - Isolation of mutations in the alpha operon of Escherichia coli that suppress the transcriptional defect conferred by a mutation in the porin regulatory gene envZ. AB - One class of mutations in the envZ gene of Escherichia coli K-12 confers a pleiotropic defect on the expression of several genes, including ompF, lamB, and phoA, that are otherwise not commonly regulated. Four second-site mutations that suppress this transcriptional defect have been isolated by using a procedure that circumvented the problem of intragenic suppressors, including true revertants. All four mutations have been mapped to the genes of the alpha operon and have been assigned tentatively to the gene rpoA, which specifies the alpha subunit of RNA polymerase. The mutations, referred to as sez (for suppressor of envZ), did not appear to confer a phenotype on an otherwise wild-type strain and did not suppress the transcriptional defects conferred by several other phenotypic classes of envZ mutations, including amber mutations. Our results led us to postulate that the alpha subunit or some other component of the alpha operon plays a role in determining the specificity of gene expression. PMID- 3031010 TI - Rhizobium meliloti insertion element ISRm2 and its use for identification of the fixX gene. AB - Two of the three plasmids of the wild-type Rhizobium meliloti 41 (pRme41a and pRme41c) carry a copy of ISRm2, a 2.7-kilobase-long transposable element. ISRm2 is terminated by 22-base-pair (bp) inverted repeat sequences, exhibiting some homology to the inverted repeats of elements generating 9-bp target sequence duplication. Transposition of ISRm2 results in a duplication of 8 bp in length, rather rare among transposable elements. DNA sequences homologous to an internal fragment of ISRm2 were found in several Rhizobium species. Transposition of ISRm2 into fixation and nodulation genes located on the symbiotic plasmid pRme41b was detected at a high frequency. Exact locations of two copies of ISRm2 which transposed into the nod-nif region on the megaplasmid were determined. In one case, integration into the protein-coding region of the hsnD gene that determines a host specificity function of nodulation occurred. In the other mutant, ISRm2 was localized upstream of nifA, where a short open reading frame coding for a new fix gene (fixX) was identified. The product of fixX is a ferredoxin carrying a characteristic cluster of cysteine residues. On the basis of the observation that the arrangement of the ISRm2 copies is identical in the free-living wild-type cells and in nitrogen-fixing nodules, we concluded that the involvement of ISRm2 transposition in the development of nitrogen-fixing symbiosis is unlikely. PMID- 3031011 TI - Specificity of mutation by UV light and delayed photoreversal in umuC-defective Escherichia coli K-12: a targeting intermediate at pyrimidine dimers. AB - Prototrophic mutants produced by UV light in Escherichia coli K-12 strains with argE3(Oc) and hisG4(Oc) defects are distinguished as backmutations and specific nonsense suppressor mutations. In strains carrying a umuC defect, mutants are not produced unless irradiated cells are incubated and then exposed to photoreversing light (delayed photoreversal mutagenesis). The mutants thus produced are found to be specifically suppressor mutations and not backmutations. The suppressor mutations are primarily glutamine tRNA ochre suppressor mutations, which have been attributed previously to mutation targeted at T = C pyrimidine dimers. In a lexA51 recA441 strain, where the SOS mutagenesis functions are constitutive, targeting at dimers is confirmed by demonstrating that the induction of glutamine tRNA suppressor mutations is susceptible to photoreversal. In the same strain induction of backmutations is not susceptible to photoreversal. Thus delayed photoreversal mutagenesis produces suppressor mutations that can be targeted at pyrimidine dimers and does not produce backmutations that are not targeted at pyrimidine dimers. This correlation supports the idea that delayed photoreversal mutagenesis in umuC defective cells reflects a mutation process arrested at a targeting pyrimidine dimer photoproduct, which is the immediate cause of both the alteration in DNA sequence and the obstruction (unless repaired) to mutation fixation and ultimate expression. PMID- 3031012 TI - Amplification of drug resistance genes flanked by inversely repeated IS1 elements: involvement of IS1-promoted DNA rearrangements before amplification. AB - Tn2653 contains one copy of the tet gene and two copies of the cat gene derived from plasmid pBR325 and is flanked by inverted repeats of IS1. Transposed onto the P1-15 prophage, it confers a chloramphenicol resistance phenotype to the Escherichia coli host. Because the prophage is perpetuated as a plasmid at about one copy per host chromosome, the host cell is still tetracycline sensitive even though P1-15 is carrying one copy of the tet gene. We isolated P1-15::Tn2653 mutants conferring a tetracycline resistance phenotype, in which the whole transposon and variable flanking P1-15 DNA segments were amplified. Amplification was most probably preceded by IS1-mediated DNA rearrangements which led to long direct repeats containing Tn2653 sequences and P1-15 DNA. Subsequent recombination events between these direct repeats led to amplification of a segment containing the tetracycline resistance gene in tandem arrays. PMID- 3031013 TI - Molecular cloning, characterization, and chromosomal localization of dapF, the Escherichia coli gene for diaminopimelate epimerase. AB - The Escherichia coli dapF gene was isolated from a cosmid library as a result of screening for clones overproducing diaminopimelate epimerase. Insertional mutagenesis was performed on the cloned dapF gene with a mini-Mu transposon, leading to chloramphenicol resistance. One of these insertions was transferred onto the chromosome by a double-recombination event, allowing us to obtain a dapF mutant. This mutant accumulated large amounts of LL-diaminopimelate, confirming the blockage in the step catalyzed by the dapF product, but did not require meso diaminopimelate for growth. The dapF gene was localized in the 85-min region of the E. coli chromosome between cya and uvrD. PMID- 3031014 TI - Characterization of the Pseudomonas aeruginosa recA analog and its protein product: rec-102 is a mutant allele of the P. aeruginosa PAO recA gene. AB - We cloned a 2.3-kilobase-pair fragment of the Pseudomonas aeruginosa PAO chromosome which is capable of complementing recA mutations of Escherichia coli. The recA-complementing activity was further localized to a 1.5-kilobase-pair PvuII-HindIII fragment. Southern blot analysis under conditions of high stringency indicated that DNA sequence homology is shared by the E. coli recA gene and the P. aeruginosa recA analog. The cloned recA analog was shown to restore resistance to methyl methanesulfonate, nitrofurantoin, and UV irradiation to E. coli recA mutants. Upon introduction of the cloned P. aeruginosa gene, these mutants regained recombination proficiency in HfrH-mediated conjugation and the ability to induce lambda prophages and SOS functions (din gene transcription) after exposure to DNA-damaging agents. Lambda prophage carrying a cI ind mutation was not inducible, suggesting that the mechanism of induction of these SOS functions by the P. aeruginosa RecA analog is similar to that by the activated E. coli RecA protein. The product of the recA analog was identified in minicells as a protein of approximately 47,000 daltons. Western blot analysis using anti-E. coli RecA antibody demonstrated that this protein is antigenically cross-reactive with the E. coli recA protein. The recA-containing fragment was cloned into the broad-host-range vector pCP13 and introduced into Rec- strains of P. aeruginosa containing the rec-102 allele. The plasmid was shown to restore recombination proficiency in FP5-mediated conjugations and to restore resistance to UV irradiation and methyl methanesulfonate to these Rec- mutants. It was shown that a wild-type allele of rec-102 is necessary for UV-mediated induction of D3 and F116 prophages. The cloned recA analog restored the UV inducibility of these prophages in rec-102 mutants. These data indicate that rec-102 is a mutant allele of the P. aeruginosa recA gene and suggest that there has been considerable conservation of the recA gene in the evolution of the gram-negative bacteria. PMID- 3031015 TI - Construction and characterization of Pseudomonas aeruginosa algB mutants: role of algB in high-level production of alginate. AB - The algB gene, which is involved in the production of alginate in Pseudomonas aeruginosa, was localized to approximately 2.2 kilobases of DNA from strain FRD by using transposon Tn501 insertion mutagenesis, subcloning, and complementation techniques. The previously reported alg-50(Ts) mutation, which confers the phenotype of temperature-sensitive alginate production, was here designated as an algB allele. A transduction-mediated gene replacement technique was used for site directed mutagenesis to isolate and characterize algB::Tn501 mutants of P. aeruginosa FRD. Although algB::Tn501 mutants had a nonmucoid phenotype (indicating an alginate deficiency), they still produced about 1 to 5% of wild type levels of alginate in most growth media and up to 16% in very rich media. The algB::Tn501 mutations had no apparent effect on growth rate or growth requirements. Using another gene replacement technique called excision marker rescue, we constructed a chromosomal algB deletion (delta algB) mutant of P. aeruginosa FRD. The delta algB mutant also produced low levels of alginate as did the algB::Tn501 mutants. The alginate produced by algB::Tn501 mutants resembled wild-type alginate by all criteria studied: molecular weight, acetylation, and proportion of mannuronic and guluronic acids. Thus, the algB gene product is apparently involved in the high-level production of alginate by P. aeruginosa and is not directly involved in the pathway leading to its biosynthesis. Chromosomal mapping of an algB::Tn501 insertion showed linkage to the trp-2 marker on the FRD chromosome as does the algB50(Ts) mutation. The excision marker rescue technique was also used to place the algB::Tn501 marker on the chromosome of characterized strains of P. aeruginosa PAO. The algB::Tn501 mutation mapped near 21 min on the PAO chromosome. PMID- 3031017 TI - Use of TnphoA to detect genes for exported proteins in Escherichia coli: identification of the plasmid-encoded gene for a periplasmic acid phosphatase. AB - The structural gene (appA) for the periplasmic acid phosphatase (optimum pH 2.5) of Escherichia coli was cloned into a plasmid by using a combination of in vivo and in vitro techniques. The position and orientation of the appA gene within the cloned DNA fragment were identified by using fusions to the alkaline phosphatase gene (phoA) generated by Tn5 IS50L::phoA (TnphoA) insertions. For TnphoA generated hybrid proteins to have high enzymatic activity, it appears that the phoA gene must be fused to a target gene coding for a signal which promotes protein export. The approach used to identify the appA gene thus appears to provide a simple general means of selectively identifying genes encoding membrane and secreted proteins. PMID- 3031018 TI - Specific magnesium-dependent diadenosine 5',5'''-P1,P3-triphosphate pyrophosphohydrolase in Escherichia coli. AB - A specific Mg2+-dependent bis(5'-adenosyl)-triphosphatase (EC 3.6.1.29) was purified 270-fold from Escherichia coli. The enzyme had a strict requirement for Mg2+. Other divalent cations, such as Mn2+, Ca2+, or Co2+, were not effective. The products of the reaction with bis(5'-adenosyl) triphosphate (Ap3A) as the substrate were ADP and AMP in stoichiometric amounts. The Km for Ap3A was 12 +/- 5 microM. Bis(5'-adenosyl) di-, tetra-, and pentaphosphates, NAD+, ATP, ADP, AMP, glucose 6-phosphate, p-nitrophenylphosphate, bis-p-nitrophenylphospate, and deoxyribosylthymine-5'-(4-nitrophenylphosphate) were not substrates of the reaction. The enzyme had a molecular mass of 36 kilodaltons (as determined both by gel filtration and sodium dodecyl sulfate-polyacrylamide gel electrophoresis), an isoelectric point of 4.84 +/- 0.05, and a pH optimum of 8.2 to 8.5. Zn2+, a known potent inhibitor of rat liver bis(5'-adenosyl)-triphosphatase and bis(5' guanosyl)-tetraphosphatase (EC 3.6 1.17), was without effect. The enzyme differs from the E. coli diadenosine 5',5'''-P1, P4-tetraphosphate pyrophosphohydrolase which, in the presence of Mn2+, also hydrolyzes Ap3A. PMID- 3031016 TI - Nascent secretory polypeptides synthesized on Escherichia coli ribosomes are not translocated across mammalian endoplasmic reticulum. AB - Cell-free protein-synthesizing systems from Escherichia coli and wheat germ were compared for their capacity to support the translocation of secretory proteins across microsomal membranes derived from mammalian endoplasmic reticulum. Three different secretory proteins, two of bacterial and one of eucaryotic origin, were tested in this respect. In all three cases a contrast between the results in the eucaryotic and procaryotic protein-synthesizing systems was revealed. Whereas the eucaryotic system, as expected, supported the translocation of nascent secretory proteins across the microsomal membranes, the procaryotic system failed to do so. This failure was not due to the absence of a translocation-promoting activity or the presence of a translocation-blocking activity in the procaryotic system. These results demonstrate a specificity in the requirement of components of the protein-synthesizing machinery for protein translocation. These components might participate in forming a functional ribosome-membrane junction during protein translocation. The nascent secretory chain alone is not sufficient for making this junction, which might involve the postulated binding of the ribosome to the signal recognition particle or another component of the membrane. PMID- 3031019 TI - Requirement of the Escherichia coli dnaA gene function for ori-2-dependent mini-F plasmid replication. AB - The mini-F plasmids pSC138, pKP1013, and pKV513 were unable to transform Escherichia coli cells with a dnaA-defective mutation under nonpermissive conditions. The dnaA defect was suppressed for host chromosome replication either by the simultaneous presence of the rnh-199 (amber) mutation or by prophage P2 sig5 integrated at the attP2II locus on the chromosome, both providing new origins for replication independent of dnaA function. The dnaA mutations tested were dnaA17, dnaA5, and dnaA46. dnaA5 and dnaA46 are missense mutations. dnaA17 is an amber mutation whose activity is controlled by the temperature-sensitive amber suppressor supF6. Under permissive conditions in which active DnaA protein was available, the mini-F plasmids efficiently transformed the cells. However, the transformants lost the plasmid as the cells multiplied under conditions in which DnaA protein was inactivated or its synthesis was arrested. As controls, plasmids pSC101 and pBR322 were examined along with mini-F; pSC101 behaved in the same manner as mini-F, showing complete dependence on dnaA for stable maintenance, whereas pBR322 was indifferent to the dnaA defect. Thus, ori-2 dependent mini-F plasmid replication seems to require active dnaA gene function. This notion was strengthened by the results of deletion analysis which revealed that integrity of at least one of the two DnaA boxes present as a tandem repeat in ori-2 was required for the origin activity of mini-F replication. PMID- 3031020 TI - Regulatory role of recF in the SOS response of Escherichia coli: impaired induction of SOS genes by UV irradiation and nalidixic acid in a recF mutant. AB - We isolated a new recF mutant of Escherichia coli K-12 by insertion of transposon Tn5 into the recF gene. This recF400::Tn5 allele displayed the same phenotypic characteristics as the classic recF143 mutation. By using Mu d(Ap lac) fusions, the induction of nine SOS genes, including recA, umuC, dinA, dinB, dinD, dinF, recN, and sulA, by UV irradiation and nalidixic acid was examined. Induction of eight genes by the two agents was impaired by recF400::Tn5 to different extents. The ninth fused SOS gene, dinF, was no longer inducible by UV when combined with recF400::Tn5. The generally impaired SOS response in recF strains did not result from weak induction of recA protein synthesis, since a recA operator-constitutive mutation did not alleviate the inhibitory effect of the recF mutation. The results suggest that recF plays a regulatory role in the SOS response. It is proposed that this role is to optimize the signal usage by recA protein to become a protease. PMID- 3031021 TI - Localization of a genetic region involved in McrB restriction by Escherichia coli K-12. AB - A 5,500-base-pair BglII-EcoRI fragment proximal to the hsd genes of Escherichia coli K-12 has been cloned in the plasmid vector pUC9. The resultant hybrid plasmid was shown to complement the mcrB mutation of E. coli K802. The presence of the hybrid plasmid in strain K802 caused an 18.3-fold drop in transformation efficiency with AluI-methylated pACYC184 relative to unmethylated pACYC184. These results indicate that the cloned DNA is involved in the McrB system restriction of 5-methylcytosine DNA. PMID- 3031022 TI - Structural inhibition and reactivation of Escherichia coli septation by elements of the SOS and TER pathways. AB - The inhibition of cell division caused by induction of the SOS pathway in Escherichia coli structurally blocks septation, as deduced from two sets of results. Potential septation sites active at the time of SOS induction became inactivated, while those initiated during the following doubling time were active. Penicillin resistance increased in wild-type UV light-irradiated cells, a behavior similar to that observed in mutants in which structural blocks were introduced by inactivation of FtsA. Potential septation sites that have been structurally blocked by either the SOS division inhibitor, furazlocillin inhibition of PBP3, or inactivation of a TER pathway component, FtsA3, could be reactivated one doubling time after removal of the inhibitory agent in the presence of an active lon gene product. Reactivation of potential septation sites blocked by the presence of an inactivated FtsA3 was significantly lower when the lon protease was not active, suggesting that Lon plays a role in the removal of inactivated TER pathway products from the blocked potential septation sites. PMID- 3031023 TI - IS2 activates the ilvA gene of Pseudomonas cepacia in Escherichia coli. AB - The ilvA gene of Pseudomonas cepacia was expressed poorly in Escherichia coli. Insertion of IS2 upstream of the cloned gene dramatically increased its transcription, resulting in an 85-fold increase in threonine dehydratase (deaminase) specific activity. The results confirm earlier reports that IS2 promotes efficient expression of foreign genes in E. coli. PMID- 3031024 TI - [Weibel-Palade bodies in endothelial cells in human dental pulp]. AB - The bodies of Weibel-Palade (BWP) were first described in 1964 in the endothelial cells of human and rat pulmonary arteries. They are probably present in the blood vessels of all human and animal tissues. Their high concentration in blood capillaries of human dental pulp makes this tissue is a good model for their study. 107 transverse pulpal sections of BWP were selected from 15 pulps. The distribution of the tubules in the matrix from regular patterns. A simple linear regression study showed that the transverse diameter of BWP is proportional to the number of contained tubules. The functions of the Von Willebrand factor or antigenic portion of factor VIII being stored in BWP of endothelial cells of other tissues is discussed. PMID- 3031025 TI - The receptor-binding sequence of urokinase. A biological function for the growth factor module of proteases. AB - Previous studies have shown that the region of human urokinase-type plasminogen activator (uPA) responsible for receptor binding resides in the amino-terminal fragment (ATF, residues 1-135) (Stoppelli, M.P., Corti, A., Soffientini, A., Cassani, G., Blasi, F., and Assoian, R.K. (1985) Proc. Natl. Acad. Sci. U.S. A. 82, 4939-4943). The area within ATF responsible for specific receptor binding has now been identified by the ability of different synthetic peptides corresponding to different regions of the amino terminus of uPA to inhibit receptor binding of 125I-labeled ATF. A peptide corresponding to human [Ala19]uPA-(12-32) resulted in 50% inhibition of ATF binding at 100 nM. Peptides uPA-(18-32) and [Ala13]uPA-(9 20) inhibit at 100 and 2000 microM, respectively. The human peptide uPA-(1-14) and the mouse peptide [Ala20]uPA-(13-33) have no effect on ATF receptor binding. This region of uPA is referred to as the growth factor module since it shares partial amino acid sequence homology (residues 14-33) to epidermal growth factor (EGF). Furthermore, this region of EGF is responsible for binding of EGF to its receptor (Komoriya, A. Hortsch, M., Meyers, C., Smith, M., Kanety, H., and Schlessinger, J. (1984) Proc. Natl. Acad. Sci. U.S.A. 81, 1351-1355). However, EGF does not inhibit ATF receptor binding. Comparison of the sequences responsible for receptor binding of uPA and EGF indicate that the region of highest homology is between residues 13-19 and 14-20 of human uPA and EGF, respectively. In addition, there is a conservation of the spacings of four cysteines in this module whereas there is no homology between residues 20-30 and 21-33 of uPA and EGF. Thus, residues 20-30 of uPA apparently confer receptor binding specificity, and residues 13-19 provide the proper conformation to the adjacent binding region. PMID- 3031026 TI - The effect of dibutyryl cyclic AMP and butyrate on F9 teratocarcinoma cellular retinoic acid-binding protein activity. AB - F9 teratocarcinoma cells contain a cellular retinoic acid-binding protein (CRABP) that may mediate the retinoic acid-induced differentiation of this cell line. Specific [3H]retinoic acid binding to CRABP in F9 stem cell cytosol is protein dependent, reaches equilibrium within 4 h at 4 degrees C, and yields 643 +/- 105 fmol of [3H]retinoic acid per mg of protein with an apparent dissociation constant of 9.2 +/- 1.1 nM. When F9 stem cells are grown in the presence of either dibutyryl cyclic AMP or sodium butyrate, CRABP activity is stimulated 2-4 fold. The effect of these drugs on CRABP activity is both time and concentration dependent, resulting in an increase in the number of binding sites for [3H]retinoic acid with no change in their affinity. The new [3H]retinoic acid binding sites have a sedimentation coefficient of 2 S and are not displaced by excess retinol. When F9 stem cells are grown in the presence of cyclic 8-bromo AMP or cholera toxin, no increase in CRABP activity is observed. We conclude that the stimulation of CRABP activity by dibutyryl cyclic AMP may result from the action of butyrate. In addition, the stimulation of retinoic acid-induced F9 cell differentiation by cyclic AMP analogs (Strickland, S., Smith, K.K., and Marotti, K.R. (1980) Cell 21, 347-355) and the inhibition of this differentiation by butyrate (Levine R. A., Campisi, J., Wang, S.-Y., and Gudas, L. J. (1984) Dev. Biol. 105, 443-450) are not correlated with increases or decreases, respectively, in the level of CRABP activity. PMID- 3031027 TI - 31P and 13C NMR studies of oxygen transfer during catalysis by 3-deoxy-D-manno octulosonate cytidylyltransferase from Escherichia coli. AB - [18O]3-Deoxy-D-manno-octulosonate (KDO), labeled at the anomeric oxygen, was prepared by exchange with [18O]H2O and used to follow the route of oxygen transfer during cytidine 5'-monophosphate-3-deoxy-D-manno-octulosonate (CMP-KDO) formation catalyzed by 3-deoxy-D-manno-octulosonate cytidylyl-transferase (CMP KDO synthetase). The 31P-NMR signal of the phosphoryl group of CMP-KDO (-5.85 ppm), which appeared as a single resonance when CMP-KDO formation took place with unenriched KDO, appeared as two peaks when CMP-KDO formation took place in the presence of a mixture of [16O]-and [18O]KDO. These results demonstrate the retention of 18O during CMP-KDO formation. Confirmation that the labeled oxygen in CMP-KDO was retained in the "bridge" position between CMP and KDO came from 13C-NMR studies of CMP-KDO formed in the presence of 90% [2-13C, 18O] KDO. The prominent C-2 KDO resonance in CMP-KDO, which is normally a doublet at 101.4 ppm (Kohlbrenner, W.E., and Fesik, S.W. (1985) J. Biol. Chem. 260, 14695-14700), appeared as four peaks when a mixture of [2-13C,16O]- and [2-13C, 18O]KDO was used, confirming the direct bonding of 18O to the C-2 of KDO in CMP-KDO. These results are consistent with a nucleophilic displacement mechanism for CMP-KDO formation. PMID- 3031028 TI - Cell surface receptor for hemopexin in human leukemia HL60 cells. Specific binding, affinity labeling, and fate of heme. AB - The binding of 125I-labeled human hemopexin to human leukemia HL60 cell at 4 degrees C was saturable with time and with increasing concentrations of 125I hemopexin. Scatchard analysis of the binding data revealed the presence of approximately 42,000 binding sites/cell with an apparent dissociation constant (Kd) of 1.0 X 10(-9) M. When cells were incubated with radioactive hemopexin at 37 degrees C, 125I-hemopexin was rapidly bound and then was dissociated after the release of heme. Treatment of surface-bound 125I-hemopexin with divalent lysine directed cross-linking disuccinimidyl suberate revealed a membrane polypeptide of about 80,000 Da, to which hemopexin is cross-linked. To examine the fate of the internalized heme, lysates from the cells previously incubated with [59Fe]heme hemopexin complex were analyzed by CM-cellulose and Sephacryl S-200 column chromatography. A considerable amount of the radioactivity was present in the fraction which co-eluted with the myeloperoxidase activity. When myeloperoxidase was isolated from the cells incubated with [59Fe]heme-hemopexin complex by immunoprecipitation with anti-myeloperoxidase antibody, radiolabeled iron associated with myeloperoxidase increased with time, and more than 30% of the radioactivity in the cells was present in the myeloperoxidase. These results indicate that the binding of hemopexin to the surface receptors triggers a release of heme and that this heme is incorporated into the intracellular myeloperoxidase. PMID- 3031029 TI - Accessibility of tryptophan residues in Na,K-ATPase. AB - The accessibility of the tryptophans in dog kidney Na,K-ATPase was studied with the technique of quenching by acrylamide. By use of a modified Stern-Volmer equation, fa, the effective fraction of tryptophans most exposed to quencher, and Ka, the effective quenching constant, were calculated. The direct Stern-Volmer plots are nonlinear under nondenaturing conditions, indicating that the tryptophan residues are unequally accessible to quencher. Modified Stern-Volmer plots revealed marked differences in the exposure of tryptophans in the E1 and E2 states. In the presence of Na or ADP, ligands that stabilize E1, these plots curve downward, indicating that the in addition to buried (unquenched) tryptophans, there is a heterogeneous class of tryptophans. In the presence of K or ouabain, conditions that favor E2, the modified Stern-Volmer plots are linear, consistent with a homogeneous population of tryptophans. Treatment with chymotrypsin to block the E1 to E2 transition results in a new set of quenching parameters which are unchanged with Na or K. Even after detergent denaturation (1% sodium dodecyl sulfate for 30 min), Stern-Volmer plots are nonlinear, and a significant fraction of tryptophan residues remain inaccessible to quencher. Denaturation with urea or guanidine HCl plus dithiothreitol increases the fraction of quenchable fluorescence even more, but still a small fraction, about 7-13%, is buried. The observed changes in exposure of the tryptophan residues would seem to account for the differences in intrinsic fluorescence seen on adding K and Na to Na,K-ATPase. The present results provide new evidence that a significant rearrangement of amino acid residues results from the E1 to E2 transition. Furthermore, a region of the molecule is inaccessible even after denaturation; this may correspond to highly hydrophobic stretches that are normally buried in the membrane. PMID- 3031030 TI - Ligand-dependent regulation of the 1,25-dihydroxyvitamin D3 receptor in rat osteosarcoma cells. AB - The specific binding of radiolabeled 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) to intact rat osteosarcoma (ROS 17/2) cells was followed for 24 h. In the presence of 0.5-1.5 nM 1,25(OH)2D3, hormone binding increased over a period of 12 h, from 1.1 X 10(4) to 1.3 X 10(5) receptors/cell. The elevated level of hormone binding persisted through 24 h provided that the initial concentration of hormone was maintained. The concentration dependence of this increase in receptor level was centered between 10 and 30 pM 1,25(OH)2D3, and the binding at 12 h exhibited the metabolite specificity expected for a 1,25(OH)2D3 receptor. The t 1/2 values for the disappearance of unoccupied and occupied receptors were roughly the same, approximately 2.7 h; therefore, the increase in hormone binding was not due to receptor stabilization. In comparison, hormone-receptor complexes appeared to dissociate with a t 1/2 of 1 h. alpha-Amanitin treatment reduced the magnitude of receptor accumulation by 50-60%, indicating that mRNA synthesis was required to achieve the maximal response. Ligand-dependent regulation of cellular receptor levels provides a mechanism for amplifying the primary hormonal signal and is predicted to influence the kinetics, magnitude, and dose dependence of cellular responses. PMID- 3031031 TI - alpha, beta-Dihydroxyisovalerate dehydratase. A superoxide-sensitive enzyme. AB - Increasing the intracellular flux of O-2 by incubating aerobic Escherichia coli with paraquat or plumbagin markedly lowered the alpha, beta-dihydroxyisovalerate dehydratase activity detectable in extracts from these cells. This effect was not seen in the absence of dioxygen and was exacerbated by inhibiting protein biosynthesis with chloramphenicol. These effects of paraquat and of plumbagin were both time- and concentration-dependent. Transfer of E. coli from aerobic to anaerobic conditions caused a rebound of the dehydratase activity, in the continued presence of paraquat and of chloramphenicol, indicating the presence of a mechanism for reactivating this enzyme. The instability of the dehydratase activity in cell extracts was exacerbated by selective removal of superoxide dismutase, but not of catalase, by immunoprecipitation. Addition of exogenous superoxide dismutase reversed the effect of immunoprecipitation; whereas catalase or inactive superoxide dismutase were ineffective. We conclude that the dehydratase is inactivated by O-2. This could account for the bacteriostatic effects of dioxygen and of paraquat. PMID- 3031032 TI - Primary structure and disruption of the phosphatidylinositol synthase gene of Saccharomyces cerevisiae. AB - The wild-type yeast nuclear gene, PIS, encodes phosphatidylinositol synthase (CDPdiacylglycerol-inositol 3-phosphatidyltransferase, EC 2.7.8.11) (Nikawa, J., and Yamashita, S. (1984) Eur. J. Biochem. 143, 251-256). We now report the sequence of the cloned 2, 129-base pair DNA and the location of the PIS coding region within the sequence. The PIS coding frame is capable of encoding 220 amino acid residues with a calculated molecular weight of 24,823. On Northern blot analysis, an RNA species that hybridized with the coding region was detected in the total poly(A)+ RNA of the wild-type yeast. The primary translation product contains a region showing local sequence homology with yeast phosphatidylserine synthase (EC 2.7.8.8) and Escherichia coli 3-phosphatidyl-1'-glycerol-3' phosphate synthase (EC 2.7.8.5), suggesting that these three enzymes are evolutionarily related. The PIS gene was disrupted in vitro through insertion of the yeast HIS3 gene into the coding region. A heterozygous diploid, PIS/pis::HIS3, constructed from a PIS/PIS his3/his3 diploid by replacing one of the wild-type PIS genes with the disrupted PIS gene, showed no segregation of viable His+ spores on tetrad analysis, indicating that disruption of the PIS gene is lethal. The nonviable spores were in an arrested state with a characteristic terminal phenotype, suggesting that the function of the PIS gene is essential for progression of the yeast cell cycle. PMID- 3031033 TI - Multiple mRNAs from rat kidney and brain encode a single Na+,K+-ATPase beta subunit protein. AB - To determine whether multiple forms of the Na+,K+-ATPase beta subunit exist in rat kidney and brain, we constructed cDNA libraries from both tissues and isolated clones carrying inserts coding for this subunit. Characterization of these cDNAs by restriction endonuclease mapping and DNA sequencing suggests that a single form of the beta subunit protein is present in these tissues. We did, however, find that the beta subunit was encoded by multiple mRNAs which can be grouped into classes based on the lengths of their 5' and 3' untranslated regions. Size variation at the 3' end is due to the use of at least two of the five potential polyadenylation sites, whereas variation at the 5' end appears to be due to the use of different initiation sites. Northern blot analysis demonstrates the presence of multiple mRNA species which correlate with the sizes observed for the different cDNA classes and also demonstrates that the larger mRNA species are present at different levels in kidney and brain. S1 and primer extension analyses suggest that the observed tissue differences in the expression of these mRNAs are due to transcription from multiple initiation sites, whose strength is tissue-dependent. In the 3' untranslated region, comparison of the nucleotide sequence of the rat beta subunit cDNAs to those from other species reveals regions of high sequence similarity. PMID- 3031034 TI - Pertussis toxin attenuates atrial natriuretic factor-mediated inhibition of adenylate cyclase. Involvement of inhibitory guanine nucleotide regulatory protein. AB - The effect of pertussis toxin treatment was studied on the inhibitory effect of atrial natriuretic factor (ANF) on adenylate cyclase activity in rat aorta. The incubation of rat aorta washed particles with pertussis toxin and [alpha-32P]NAD resulted in the ADP-ribosylation of a single 40-kDa protein. In addition, pertussis toxin treatment enhanced guanosine 5'-O-(thiotriphosphate) and isoproterenol-stimulated adenylate cyclase activities and attenuated the ANF mediated inhibition of basal, isoproterenol-, and forskolin-stimulated adenylate cyclase activities. These data suggest that ANF receptors are coupled to adenylate cyclase through inhibitory guanine nucleotide regulatory protein. PMID- 3031035 TI - Biochemical changes during sucrose deprivation in higher plant cells. Phosphorus 31 nuclear magnetic resonance studies. AB - An experimental arrangement was described that enables nuclear magnetic resonance spectra of compressed plant cells to be recorded while circulating a medium through the sample. The system provided a convenient arrangement for monitoring by 31P NMR the behavior of plant cells over a long period of time under different conditions such as sucrose starvation. Perfusion of compressed sycamore cells with sucrose-free culture medium triggered a progressive decrease in the glucose 6-P and uridine-5'-diphosphate-alpha-D-glucose resonances over 30 h. When almost all the intracellular carbohydrate pool had disappeared the nucleotide triphosphate resonances decline progressively. These changes were accompanied by a Pi accumulation in the vacuole and a phosphorylcholine (P-choline) accumulation in the cytoplasm. The very long lag phase observed for ATP and P-choline evolution was comparable with that observed for the progressive intracellular digestion of cytoplasmic constituents (Journet, E., Bligny, R. and Douce, R. (1986) J. Biol. Chem. 261, 3193-3199). Addition of sucrose in the circulating system after a long period of sucrose starvation led to a disappearance of the cytoplasmic Pi resonance and a marked increase in that of glucose 6-P. Under these conditions the vacuolar Pi pool did not fluctuate to buffer the Pi in the cytoplasm. The results suggest that Pi which has been sequestered in the vacuole during the course of sucrose starvation is not restored to the cytoplasm for rapid metabolic processes. Furthermore, the presence of P-choline in plant cells in large excess should be considered as a good marker of membrane utilization after a long period of sucrose starvation and is very likely related to stress. PMID- 3031036 TI - Guanine nucleotides induce Ca2+-independent insulin secretion from permeabilized RINm5F cells. AB - The role of guanine nucleotides in insulin secretion was investigated in electrically permeabilized RINm5F cells. Ca2+ stimulated insulin release (EC50 approximately 2 microM Ca2+). The GTP stable analog, GTP gamma S, elicited insulin secretion at vanishingly low Ca2+ concentrations (less than 10(-11) M), slightly potentiated the response to intermediate Ca2+ levels, but exerted less than additive effects at maximal Ca2+ concentrations. The GDP analog, GDP beta S, inhibited both GTP gamma S- and Ca2+-stimulated secretion. The action of GTP gamma S was not mediated by cAMP, as the latter only enhanced Ca2+-induced secretion. In contrast, 12-O-tetradecanoylphorbol-13-acetate, an activator of protein kinase C, promoted insulin release at nonstimulatory Ca2+ levels as well as potentiating the Ca2+ response. GTP analogs stimulated hydrolysis of phosphatidylinositol 4,5-bisphosphate (PtdInsP2), as assessed by inositol phosphate generation. However, this could not fully explain guanine nucleotide induced secretion because: GTP gamma S-stimulated PtdInsP2 breakdown was totally dependent on Ca2+ and abolished at Ca2+ below 10(-11) M; at these Ca2+ levels, activators of protein kinase C were weak or ineffective secretagogues; the GTP analog Gpp(NH)p was much less effective than GTP gamma S in activating PtdInsP2 hydrolysis, while fully mimicking the effect on Ca2+-independent secretion. Both GTP gamma S-induced PtdInsP2 hydrolysis and insulin release were insensitive to pertussis toxin and cholera toxin. The findings point to a guanine nucleotide regulated site in the activation of insulin secretion different from the known transmembrane signalling systems. PMID- 3031037 TI - Angiotensin II-stimulated Na+/H+ exchange in cultured vascular smooth muscle cells. Evidence for protein kinase C-dependent and -independent pathways. AB - Angiotensin II, a potent vasoconstrictor, is known to stimulate Ca2+ mobilization and Na+ influx in vascular smooth muscle cells (VSMC). The fact that the Na+/H+ exchange inhibitor, amiloride, blocks angiotensin II-stimulated Na+ influx and is itself a vasodilator suggests that Na+/H+ exchange may play a role in the angiotensin II-mediated effects on VSMC. We have used a pH-sensitive fluorescent dye to study Na+/H+ exchange in cultured rat aortic VSMC. Basal intracellular pH was 7.08 in physiological saline buffer. Angiotensin II stimulation caused an initial transient acidification, followed by a Na+-dependent alkalinization. Angiotensin II increased the rate of alkalinization with apparent threshold, half maximal, and maximal effect of 0.01, 3, and 100 nM, respectively. Angiotensin II stimulation appeared to be mediated by a shift in the Km of the Na+/H+ exchanger for extracellular Na+. Since angiotensin II activates phospholipase C in VSMC, we tested the possibility that angiotensin II increased Na+/H+ exchange by activation of protein kinase C via stimulation of diacylglycerol formation. The phorbol ester, 12-O-tetradecanoylphorbol-13-acetate (TPA), stimulated Na+/H+ exchange in VSMC cultured for 24 h in serum-free medium, and the subsequent angiotensin II response was inhibited. However, VSMC grown in serum and treated for 24 h with TPA to decrease protein kinase C activity showed no inhibition of angiotensin II-stimulated Na+/H+ exchange. TPA caused no intracellular alkalinization of VSMC grown in serum, while the angiotensin II response was actually enhanced compared to VSMC deprived of serum for 24 h. We conclude that angiotensin II stimulates an amiloride-sensitive Na+/H+ exchange system in cultured VSMC which is mediated by protein kinase C-dependent and -independent mechanisms. Angiotensin II-mediated Na+ influx and intracellular alkalinization may play a role in excitation-response coupling in vascular smooth muscle. PMID- 3031039 TI - Angiotensin II effect on cytosolic pH in cultured rat vascular smooth muscle cells. AB - This study investigated fluctuations of cytosolic pH (pHi) of cultured rat vascular smooth muscle cells (VSMCs) in reaction to metabolic alterations induced by angiotensin II (AII). Serially passed VSMCs from Wistar rat aortae were grown on coverslips and loaded with the pH-sensitive fluorescent indicator 2',7' bis(carboxyethyl)-5,6-carboxyfluorescein. A biphasic reaction was seen after exposure of these cells to AII (1 nM to 1 microM); an initial and relatively brief phase of acidification was followed by sustained alkalinization. The rate of acidification and magnitude of alkalinization were dose-dependent. This biphasic effect of AII was also demonstrated in Ca2+-free medium and was mimicked by subjecting VSMCs to the calcium ionophore A23187 (5 microM) in Ca2+-containing medium but not in Ca2+-free medium. Verapamil (10 microM) almost entirely eliminated the AII-induced acidification, whereas amiloride analogues 5-(N-methyl N-isobutyl)amiloride and 5-(N-ethyl-N-isopropyl)amiloride (100 microM) as well as Na+-deficient medium abolished the subsequent (alkalinization) phase produced by the hormone. Activation of the Na+/H+ antiport by subjecting VSMCs to phorbol 12 myristate 13-acetate (100 nM) prevented a subsequent effect of AII on the pHi profile. This resistance to a further action of the hormone was not mediated via cytoplasmic alkalinization. AII produced a dramatic redistribution in the cellular compartments of 45Ca2+ associated with accelerated 45Ca2+ washout. These findings suggest that the AII-induced acidification phase may relate to activation of the Ca2+ pump (Ca2+/H+ exchange) and that this process can take place in the presence and absence of extracellular Ca2+. The alkalinization phase is the consequence of stimulation of the Na+/H+ antiport, which in cultured VSMCs can be activated by a rise in cytosolic free Ca2+ as well as other mechanisms. PMID- 3031038 TI - Early agonist-mediated ionic events in cultured vascular smooth muscle cells. Calcium mobilization is associated with intracellular acidification. AB - Angiotensin II, a potent vasoconstrictor peptide, increases free cytoplasmic Ca2+ concentration ([Ca2+]i) in vascular smooth muscle cells (VSMC) by release of nonmitochondrial Ca2+ stores and stimulates an amiloride-sensitive Na+ influx, presumably via Na+/H+ exchange. We recently have found that the angiotensin II mediated change in VSMC intracellular pH has two components, an early rapid acidification phase and a slower recovery phase involving Na+-dependent alkalinization. In the present study, we show that the early acidification is not mediated via Na+/H+ exchange. Instead, we propose a mechanism which involves increases in [Ca2+]i and Ca2+ efflux with a subsequent rise in intracellular H+. Agonists, in addition to angiotensin II, which increase [Ca2+]i in cultured VSMC, including platelet-derived growth factor, vasopressin, and bradykinin, induce an acidification, while agonists which fail to raise [Ca2+]i do not. The time course and magnitude of agonist-stimulated 45Ca2+ efflux correlate with the acidification response. The angiotensin II concentration-response relationship for acidification and Ca2+ mobilization are similar. Furthermore, inhibition of changes in [Ca2+]i by treatment with phorbol ester, cyclic GMP, or quin2 loading prevent agonist-mediated acidification. The effects of altering extracellular [Ca2+] and [H+] on agonist-mediated intracellular acidification and H+ efflux suggest that the acidification is due to ATP-dependent unidirectional H+ influx, perhaps via the plasma membrane Ca2+-ATPase, and not to a Ca2+/H+ antiport. This agonist-mediated acidification represents a previously undescribed ionic event in VSMC activation which may be involved in excitation-response coupling. PMID- 3031040 TI - The human IL-2 receptor gene contains a positive regulatory element that functions in cultured cells and cell-free extracts. AB - The 130-base pair fragment located between 220 and 90 base pairs upstream of the major transcription initiation site of the human interleukin 2 (IL-2) receptor gene had positive regulatory effect on the early promoter of simian virus 40 as well as its own promoter. This fragment seems to be responsible for not only cell specific but also lymphokine-induced expression of the IL-2 receptor gene as assessed by DNA transfection. The same DNA fragment directed cell-specific transcription of the IL-2 receptor gene in extract of HTLV-I-infected T cells, MT 1, but not of Epstein-Barr virus-transformed B cells, CESS. The addition of small amounts of MT-1 extract to CESS extract resulted in specific expression of the IL 2 receptor gene, indicating that cell-specific expression is regulated by trans acting molecules in MT-1 extract. PMID- 3031041 TI - Oxidation-reduction properties of the two Fe4S4 clusters in Clostridium pasteurianum ferredoxin. AB - The ferredoxin from Clostridium pasteurianum contains two Fe4S4 clusters. In this paper we determine their oxidation-reduction midpoint potentials; we find them to be essentially identical (within 10 mV) and to have pH-independent Em values of 412 +/- 11 mV from pH 6.3 to 10.0. PMID- 3031042 TI - Purification and properties of the catalytic subunit of the branched-chain alpha keto acid dehydrogenase phosphatase from bovine kidney mitochondria. AB - The catalytic subunit of the branched-chain alpha-keto acid dehydrogenase (BCKDH) phosphatase (Damuni, Z., Merryfield, M.L., Humphreys, J.S., and Reed, L.J., (1984) Proc. Natl. Acad. Sci. U.S.A. 81, 4335-4338) has been purified over 50,000 fold from extracts of bovine kidney mitochondria. The apparently homogeneous protein consists of a single polypeptide chain with an apparent Mr = approximately 33,000 as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. BCKDH phosphatase, with an apparent Mr = 460,000, was dissociated to its catalytic subunit with no apparent change in activity, at an early stage in the purification procedure by treatment with 6 M urea. The specific activity of the catalytic subunit was 1,500-2,500 units/mg. The catalytic subunit exhibited approximately 10% maximal activity with 32P-labeled pyruvate dehydrogenase complex but was inactive with phosphorylase a and with p nitrophenyl phosphate. The catalytic subunit, like the Mr = 460,000 species, was inhibited by nanomolar concentrations of BCKDH phosphatase inhibitor protein, was unaffected by protein phosphatase inhibitor 1 and inhibitor 2, and was inhibited by nucleoside tri- and diphosphates but not by nucleoside monophosphates. PMID- 3031045 TI - The mechanism of utilization of polyphosphate by polyphosphate glucokinase from Propionibacterium shermanii. AB - The phosphorylation of glucose by polyphosphate glucokinase with both long- and short-chain polyphosphates has been shown to occur by either a nonprocessive mechanism, i.e. with repeated association and dissociation of the polyphosphate from the enzyme after each phosphorylation or by a quasiprocessive mechanism in which several phosphorylations occur prior to the release of polyphosphate and the reassociation with the enzyme. In contrast, the phosphorylation of ADP to ATP by polyphosphate kinase is by a strictly processive mechanism; the phosphorylation occurs without release of the polymer from the enzyme prior to termination of the reaction (Robinson, N. A., Clark, J. E., and Wood, H. G. (1987) J. Biol. Chem. 262, 5216-5222). The demonstration that the mechanism is quasi-or nonprocessive was accomplished by electrophoresis using a variety of concentrations of polyacrylamide gels which made it possible to detect the intermediate sizes formed during the reactions. It also has been shown that all chains longer than about 100 are used simultaneously, but with chains of less than 100 residues, there is preferential utilization of the longest chains. Thus a narrow range of sizes is formed from a heterogeneous mixture of long chains. It is this formation of the narrow range of sizes that makes it possible to use polyphosphate glucokinase for the determination of the average size of long chains (Pepin, C. A., Wood, H. G., and Robinson, N. A. (1986) Biochem. Int. 12, 111-123). PMID- 3031044 TI - Polyphosphate kinase from Propionibacterium shermanii. Demonstration that polyphosphates are primers and determination of the size of the synthesized polyphosphate. AB - Polyphosphate kinase from Propionibacterium shermanii was purified to 70% homogeneity and shown to be a monomeric enzyme of molecular weight 83,000 +/- 3,000. It was demonstrated that short chains of polyphosphate serve as primers by using [32P]polyphosphate, 6-80 residues in length for synthesis of long-chain polyphosphate glucokinase, the radiolabel was found to be at the end of the polymer, proving that the mechanism of elongation of polyphosphate by polyphosphate kinase is strictly processive. Only 1 out of 3-8 of the polyphosphate chains contained the primer, indicating that there is a second unknown pathway of initiation which does not involve the polyphosphate primer. The termination of polyphosphate synthesis was investigated. With polyphosphate as a primer, the majority of the synthesized polyphosphate was 750 residues in length. With phosphate, in place of the polyphosphate primer, the major portion was about 2,000 residues in length but there was a large span of chain lengths down to 300. Termination is influenced by pH, temperature, and the concentration of the polyphosphate primer, with the chain length decreasing as either the temperature or the concentration of primer is increased. PMID- 3031043 TI - Purification and characterization of a divalent cation-independent, spermine stimulated protein phosphatase from bovine kidney mitochondria. AB - A divalent cation-independent and spermine-stimulated phosphatase (protein phosphatase SP) that is active toward the phosphorylated pyruvate dehydrogenase complex has been purified about 15,000-fold to near homogeneity from extracts of bovine kidney mitochondria. Half-maximal stimulation, 1.5- to 3-fold at pH 7.0 7.3, occurred at 0.5 mM spermine. Protein phosphatase SP exhibited an apparent Mr = 140,000-170,000 as estimated by gel-filtration chromatography on Sephacryl S 300. Two major subunits, with apparent Mr = 60,000 and 34,000, were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Gel-permeation chromatography of protein phosphatase SP on Sephacryl S-200 in the presence of 6 M urea and 1.4 M NaCl increased its activity 3- to 6-fold and was accompanied by conversion to the catalytic subunit with an apparent Mr = approximately 34,000. Protein phosphatase SP was inactive with p-nitrophenyl phosphate and was not inhibited by protein phosphatase inhibitor 1, inhibitor 2, or the protein inhibitor of branched-chain alpha-keto acid dehydrogenase phosphatase. Protein phosphatase SP was inhibited by sheep antibody to the catalytic subunit of protein phosphatase 2A from rabbit skeletal muscle. It appears that protein phosphatase SP is related to protein phosphatase 2A. PMID- 3031046 TI - Ganglioside-modulated protein phosphorylation. Partial purification and characterization of a ganglioside-stimulated protein kinase in brain. AB - Gangliosides have profound modulatory effects on protein phosphorylation in brain. A protein kinase activated directly by gangliosides has been partially purified from the particulate fractions of guinea pig brain through extraction with nonionic detergent, ion-exchange chromatography, hydrophobic chromatography, and gel filtration. This novel ganglioside-stimulated protein kinase is distinct from cAMP-dependent, Ca2+/calmodulin-dependent, and Ca2+/phospholipid-dependent protein kinases. The partially purified kinase preparation could undergo ganglioside-stimulated autophosphorylation of a major phosphoprotein with Mr corresponding to 68,000. It also could phosphorylate exogenous substrates such as the synthetic peptide Leu-Arg-Arg-Ala Ser-Leu-Gly. The requirement of gangliosides for the activation of kinase activity is dose-dependent and specific. Among the various gangliosides tested, GT1b and GD1a were found to be the most potent activators, whereas GD1b and GM1 were slightly less effective. The activation process is rapid and does not require the presence of Ca2+, suggesting that the stimulatory effect of gangliosides is not mediated through limited proteolysis or Ca2+-glycolipid complexes. Although the exact physiological significance of the ganglioside-stimulated protein kinase is not known at present, it is possible that certain functions related to gangliosides in the nervous system are mediated through the activation of this novel enzyme. PMID- 3031047 TI - Tissue-specific and periodic changes in the nuclease sensitivity of the fibroin gene chromatin in the silkworm Bombyx mori. AB - Nuclei of substantial purity were isolated from the middle or posterior silk glands of the silkworm Bombyx mori larvae. Both the fibroin H- and L-chain gene sequences in the isolated nuclei from the posterior silk glands of the fifth instar larvae, where the genes are transcribed actively, are extremely sensitive to the digestion with DNaseI; on the other hand, these sequences in the middle silk gland nuclei from the same larvae, where the genes are not expressed, are markedly resistant to the digestion. The H-chain gene sequences in the posterior silk gland nuclei from the fifth instar larvae are also highly susceptible to the digestion with micrococcal nuclease, HinfI, and HhaI. The digestion products with micrococcal nuclease show a continuous size distribution. The H-chain gene sequences in the middle silk gland nuclei or the posterior silk gland nuclei from the fourth molting stage are cleaved partially into nucleosome dimer to oligomer sizes upon digestion with higher concentrations of micrococcal nuclease, suggesting that the inactive forms of the H-chain gene chromatin are constructed by folding of the chromatin fiber containing a regular array of nucleosomes. Hypersensitive sites to micrococcal nuclease are present near both ends of the second exon, a major body of the fibroin H-chain gene, in both the active and inactive forms of the chromatin. The DNaseI or micrococcal nuclease sensitivity of the H-chain gene chromatin in the posterior silk gland nuclei shows periodical changes corresponding to the intermolt-molt-intermolt cycle. PMID- 3031048 TI - Molecular characterization of the cell cycle-regulated thymidylate synthase gene of Saccharomyces cerevisiae. AB - The complete nucleotide sequence of a 1.8-kilobase DNA fragment containing the cell cycle-regulated thymidylate synthase gene (TMP 1) of the yeast Saccharomyces cerevisiae is presented. This analysis has revealed a 912-base pair open reading frame which encodes a 304-amino acid residue protein with a calculated Mr of 35,007. The tmp1-6 and cdc21-1 mutant alleles of this gene also have been sequenced, and both show single base pair changes which would result in different amino acid substitutions. The amino acid sequence of the yeast thymidylate synthase gene derived from the DNA sequence shows considerable homology when compared with the human, mouse, Herpesvirus saimiri, Leishmania major, Leishmania tropica, Escherichia coli, Lactobacillus casei, bacteriophage T4, and Bacillus subtilis phage phi 3T enzymes. Northern blot hybridization reveals that the TMP 1 mRNA is a 1.15-kilobase polyadenylated transcript. A set of consensus yeast mRNA splice sequences appears within the open reading frame of TMP 1, but S1 nuclease protection experiments reveal that splicing of the mRNA does not occur. Disruption of the gene by the introduction of a large insertion did not produce any defect besides the expected dependence on dTMP for growth. Specifically, the viability of the mutants in the presence of dTMP indicates that the protein does not play a significant structural role in a complex of replication enzymes. PMID- 3031049 TI - Interstitial cell heterogeneity in rat testes. I. Purification of collagenase dispersed Leydig cells by unit gravity sedimentation and demonstration of binding sites for gonadotropin in light cells versus enhanced steroidogenesis in heavier cells. AB - Two testicular interstitial cell fractions, light and heavier, biochemically and morphologically distinct were obtained by a unit gravity sedimentation procedure. Binding sites for 125I-labeled human chorionic gonadotropin (hCG) were preferentially localized in the light cell fraction (apparent Kd = 2.02 X 10(-10) M; Bmax = 1.17 X 10(-5) nmol/2 X 10(6) cells). These cells did not synthesize testosterone in response to hCG, but the basal release of testosterone was higher than by cells in the heavier fraction (2.49 +/- 0.02 ng/2 X 10(6) cells in the light versus 0.22 +/- 0.00 ng/2 X 10(6) cells in the heavier fraction). The cells in the heavier fraction bound little or no hCG. The binding data from this fraction did not obey saturation kinetics, but testosterone levels were elevated 700-800% in the presence of hCG (i.e. basal value 0.22 +/- 0.00 ng/2 X 10(6) cells versus 1.81 +/- 0.04 ng/2 X 10(6) cells in hCG-stimulated cells). Electron microscopy revealed that heavier cells had features typical of Leydig cells such as large ovoid nucleus with peripherally located heterochromatin, numerous mitochondria with tubular cristae, some lipid droplets, extensively developed smooth endoplasmic reticulum, and well developed Golgi complex. The cells in the light fraction contained an ovoid nucleus with one or more deep infoldings, and their most notable cytoplasmic feature was the presence of numerous vacuoles of varying sizes and shapes. Based upon this and the investigation which follows (Bhalla, V.K., Flasch, M.V., Browne, E.S., Sohal, G.S., and Sharawy, M.M. (1987) J. Biol. Chem. 262, 5322-5332), we conclude that occupancy of high affinity hCG binding sites, generally assumed to be coupled to steroidogenesis, is not necessarily related to the elicitation of this biological response. PMID- 3031050 TI - Interstitial cell heterogeneity in rat testes. II. Purification of cells by Percoll and metrizamide gradient centrifugation with preferential localization of gonadotropin binding sites in light cell fraction and hormone-induced steroidogenesis in heavier cell fraction. AB - The ability of 125I-labeled human chorionic gonadotropin (125I-labeled hCG) to bind and stimulate steroidogenesis was studied in light cells (density, 1.053 1.065 g/cm3) and heavier cells (density, 1.090-1.110 g/cm3) purified from collagenase-dispersed rat testicular interstitial cells by unit gravity sedimentation (Bhalla, V.K., Rajan, V.P., Burgett, A.C., and Sohal, G.S. (1987) J. Biol. Chem. 262, 5313-5321). Preferential localization of gonadotropin binding sites was demonstrated on light cells, and the heavier cells produced testosterone in response to hCG without occupancy of high affinity (Kd = 2.02 X 10(-10) M) binding sites. In this study, established methods for interstitial cell purification involving gradient centrifugation were utilized to demonstrate the cell heterogeneity. Light cells bound hCG with high affinity (Kd = 3 X 10( 10) M) without manifestation of steroidogenic response. The heavier cells responded to hCG with elicitation of steroidogenesis, but the occupancy was negligible. Stimulation of steroidogenesis by hCG in heavier cells was dose and time dependent. Dibutyryl and bromo cyclic AMP (1 mM) also promoted steroidogenesis comparable to a level stimulated by the tropic hormone (700% stimulation). The concept of spare receptors was tested in purified cell fractions. Upon cell purification, no saturable high affinity binding sites were observed in the heavier cell fraction. Autoradiographic analyses at the electron microscopical level supported this conclusion. Our data suggest that target cell activation is not preceded by hormone occupancy of high affinity binding sites. A model for defining the functional domains of the physiological receptor for hCG is presented. PMID- 3031051 TI - Mapping the active site tyrosine of Escherichia coli DNA gyrase. AB - We have identified tyrosine 122 of the A subunit of Escherichia coli DNA gyrase as the tyrosine that becomes covalently bound to DNA when the enzyme breaks the phosphodiester bonds of DNA. The covalent gyrase X DNA complex was isolated following cleavage of the DNA by gyrase in the presence of the gyrase inhibitor oxolinic acid. The active site tyrosine was first mapped to two overlapping peptides. Its precise position in the sequence of the A subunit of gyrase was then determined by sequencing of a peptide bound to DNA. We also present a method for mapping sites of DNA attachment in a protein of known amino acid sequence. The covalent complex of DNA and protein is treated with proteases that cut specifically. The electrophoretic mobilities of the resulting peptide-bound DNA molecules are correlated with the sizes of the bound peptides, allowing determination of the site of attachment of the DNA. PMID- 3031052 TI - The enhancer of the immunoglobulin heavy chain locus is flanked by presumptive chromosomal loop anchorage elements. AB - We have located presumptive chromosomal loop anchorage elements within the mouse heavy chain immunoglobulin locus. Analysis of 31 kilobases spanning diversity, joining, enhancer, switch, and the mu and delta constant regions reveals that only a single 1-kilobase segment exhibits specific binding to nuclear matrices. It is of particular significance that the transcriptional enhancer element resides within this matrix association region (MAR). Fine structure mapping indicates that binding is mediated by A+T-rich approximately 350-base pair segments that reside on either side of the enhancer. The MAR sequences residing 5' of the enhancer contain topoisomerase II consensus sequences like the MAR located upstream of the kappa light chain gene enhancer. The heavy chain gene MARs, however, exhibit a lower affinity for matrix association compared to the kappa gene MAR. Significantly, the juxtaposition of enhancer elements with MARs appears to be evolutionarily conserved within the immunoglobulin genes, suggesting that MARs may act as positive and/or negative regulators of enhancer function. PMID- 3031053 TI - Block duplications of a zeta-zeta-alpha-theta gene set in the rabbit alpha-like globin gene cluster. AB - In order to understand the coordinate regulation between the alpha-like and beta like globins during the developmental switches in hemoglobin synthesis, we have studied the rabbit alpha-like globin gene family. A cluster of six linked genes arranged 5'-zeta 1-alpha 1-theta 1-zeta 2-zeta 3-theta 2-3' has been isolated as a set of overlapping clones from a library of rabbit genomic DNA. Blot hybridization analysis of genomic DNA not only confirms this linkage arrangement but also reveals the presence of additional zeta and theta genes. We propose that this gene cluster was generated by a block duplication of a set of alpha-like genes; the proposed duplication unit is zeta-zeta-alpha-theta. Further duplications of a zeta-zeta-theta set are also proposed to have occurred. As expected for a duplicated locus, the rabbit alpha-like gene cluster contains long blocks of internal homology. The Z homology block is about 7.2 kilobase pairs long and contains the zeta genes; the T homology block is about 4.7 kilobase pairs long and contains a theta gene. Surprisingly, both Z and T homology blocks are flanked by a common junction sequence (J) which contains a region very similar to the 3'-untranslated sequence of an alpha-globin gene. Analysis of the J sequences suggests a recombination mechanism by which the alpha gene could have been deleted from the second set of genes in the cluster (zeta 2-zeta 3-theta 2). The relationships among the genes in characterized alpha-like gene clusters in mammals are summarized. The rabbit gene cluster differs from those of other mammals principally in the loss of a gene orthologous to the human psi alpha 1 and in the block duplication of the zeta-zeta-alpha-theta gene set. PMID- 3031054 TI - Synthesis and secretion of serum gelsolin by smooth muscle tissue. AB - Gelsolin is one of many actin binding proteins which regulate the structure of intracellular microfilaments. A secretory form of gelsolin, a protein also known as "actin depolymerizing factor" or "brevin," is present in animal sera. In the present studies, we: demonstrate that a 90-kDa secretory protein produced by chicken gizzard smooth muscle is serum gelsolin; show that chicken serum gelsolin, as compared with its mammalian counterparts, lacks 26 amino acid residues at its NH2-terminal end; show that gizzard smooth muscle devotes on the order of 100 times more of its total protein synthetic effort (about 1% of the total) to the production of serum gelsolin than does liver, a previously speculated major source of this protein; and give evidence that rat tissues which are rich in smooth muscle cells (blood vessels, uterine muscle) also produce serum gelsolin. Our work suggests that, in vivo, smooth muscle-containing tissues may be major producers of the serum form of this actin binding protein. PMID- 3031055 TI - Import of hybrid vesicular stomatitis G protein to the mitochondrial inner membrane. AB - cDNA fusions were employed to construct a 35-kDa hybrid protein bearing the amino terminal signal sequence of pre-ornithine carbamoyltransferase (pOCT), a mitochondrial matrix enzyme, fused to the carboxyl-terminal half of vesicular stomatitis virus G protein (VSV G). Following transcription-translation in vitro, the hybrid protein was imported by purified mitochondria and processed at its amino terminus by the matrix processing enzyme. The protein, however, remained anchored in the mitochondrial inner membrane, apparently in a transmembrane configuration, via the hydrophobic VSV G stop-transfer domain; a small portion (approximately 2 kDa) of the G protein fragment was accessible to protease digestion only after selective permeabilization of the mitochondrial outer membrane with digitonin, a finding consistent with localization of the extreme carboxyl-terminal cytoplasmic tail of G in the intermembrane space. The results demonstrate that the membrane-anchor domain of VSV G can function in a post translational manner and can operate in membranes other than those derived from the endoplasmic reticulum. However, it appears to be selectively recognized as a stop-transfer signal by the translocation machinery of the mitochondrial inner, rather than outer, membrane. PMID- 3031056 TI - Identification of a receptor for peptides of the bombesin family in Swiss 3T3 cells by affinity cross-linking. AB - The cross-linking agent ethylene glycol-bis(succinimidyl succinate) was used to covalently link 125I-labeled gastrin releasing peptide (125I-GRP) to an Mr 75,000 85,000 surface protein in Swiss 3T3 cells that displays many characteristics of a specific receptor for peptides of the bombesin family. This protein was not present in other cell lines which do not exhibit receptors for bombesin-like peptides. Unlabeled GRP competed for affinity labeling of the Mr 75,000-85,000 protein in a concentration-dependent manner, and other bombesin-related peptides also inhibited the cross-linking of 125I-GRP to this component. In contrast, high concentrations of a variety of other peptide hormones and mitogens had no effect. Affinity labeling of the Mr 75,000-85,000 protein was dependent on the concentration of 125I-GRP and exhibited saturability. 125I-GRP affinity labeling of this protein was also demonstrated by two-dimensional gel electrophoresis. These studies suggest that an Mr 75,000-85,000 surface protein with an isoelectric point of 6.0 to 6.5 is a major component of the receptor for peptides of the bombesin family in Swiss 3T3 cells. PMID- 3031057 TI - Structure, organization, and regulation of a hamster proline-rich protein gene. A multigene family. AB - Genomic DNA fragments bearing proline-rich protein (PRP) genes expressed specifically in hamster parotid glands have been isolated and characterized. Complete exonic sequences as well as intronic and a considerable portion of the flanking sequences are reported for a PRP gene, H29. H29 is interrupted by three intervening sequences, with consensus splice junctions, and it likely encodes the acidic hamster PRP Hp43a. Exceedingly high homology of the 5'-untranslated region and the sequence encoding the signal peptide is observed with other PRPs of all species studied. Significant homology was also detected among the repetitive sequences of the mature acidic PRPs from human, mouse, hamster, and rat. This conservation of the internal repeats of the PRPs suggested that proline-rich protein gene evolution involved intragenic duplication of internal repeats and gene duplication and conversion. Both hamster and mouse PRP genes (H29 and mouse proline-rich protein gene, respectively) share considerable sequence similarity in the 5'-flanking regions for about 100 base pairs upstream. The remainder of the upstream sequences were heterologous except for three oligonucleotide regions with 60-70% sequence conservation. These three regions are thought to be involved in the regulation of the tissue-specific PRP gene induction. PMID- 3031058 TI - Alpha-melanocyte-stimulating hormone regulation of tyrosinase in Cloudman S-91 mouse melanoma cell cultures. AB - alpha-MSH (melanocyte-stimulating hormone) causes an increase in tyrosinase activity (O-diphenol-O2 oxidoreductase; EC 1.14.18.1) in Cloudman S-91 mouse melanoma cell cultures following a lag period of approximately 9 h. Treatment of cells with 2 X 10(-7)M alpha-MSH for 6 days results in a 90-fold increase in the specific activity of the enzyme. The hormone-mediated increase in tyrosinase activity is dependent upon continued transcription since the enzyme induction is suppressed by either cordycepin (1 microgram/ml) or alpha-amanitin (10 micrograms/ml). Immunoprecipitation analysis of pulse-labeled tyrosinase from control and MSH-treated cultures (48-h exposure) has demonstrated that MSH stimulates tyrosinase synthesis by approximately 4-fold, a level of induction which does not correspond to the observed 14-fold increase in enzyme activity. When immunotitration curves were developed from cell extracts of control and MSH treated cultures using immunoprecipitation and competitive enzyme-linked immunosorbent assay protocols, evidence for the presence of immunologically active but catalytically less active enzyme in untreated melanoma cell cultures was demonstrated. Degradation rates of tyrosinase were found to be similar in control cultures or in cells treated with MSH for up to 48 h. Taken together, these results suggest that in addition to stimulating tyrosinase synthesis, MSH may also promote an increase in the catalytic efficiency of the enzyme. PMID- 3031059 TI - Identification and characterization of leukotriene D4 receptors and signal transduction processes in rat basophilic leukemia cells. AB - The leukotriene D4 (LTD4) receptor on rat basophilic leukemia (RBL-1) cell membranes was characterized using a radioligand binding assay. [3H]LTD4 binding to RBL-1 membrane receptors was stereoselective, specific, and saturable. The binding affinity and maximum binding density of [3H]LTD4 to RBL-1 membrane receptors were 0.9 +/- 0.2 nM and 800 +/- 125 fmol/mg protein, respectively. Binding of [3H]LTD4 to the receptors was enhanced by divalent cations (Ca2+, Mg2+, and Mn2+) and inhibited by guanine nucleotides and sodium ions, specifically, indicating that a guanine nucleotide-binding protein may regulate the agonist-receptor interaction. LTD4, LTE4 agonist and antagonist analogs competed with the radioligand in binding to the RBL-1 LTD4 receptors. The binding affinities of these analogs correlated with (a) those determined from the guinea pig lung LTD4 receptors and (b) the pharmacological activities in smooth muscle contraction. LTD4 and related agonists also induced time- and concentration dependent phosphatidylinositol hydrolysis in RBL-1 cells. The LTD4 induction of inositol 1-phosphate was potent, stereoselective, specific, and was blocked by LTD4 receptor antagonists. The rank order potency of agonist-induced inositol 1 phosphate formation in RBL-1 cells was equivalent to the receptor binding affinity determined using either RBL-1 cell or guinea pig lung membranes. These studies have demonstrated the G protein coupled LTD4 receptors on RBL-1 cell membranes. Binding of agonists to the receptor may activate the G protein regulated phospholipase C to induce hydrolysis of phosphatidylinositol. The hydrolytic products of phosphatidylinositol, possibly inositol trisphosphate and diacylglycerol, may be the intracellular messengers for LTD4 receptors in RBL-1 cells. PMID- 3031060 TI - NADPH oxidase of human neutrophils. Subcellular localization and characterization of an arachidonate-activatable superoxide-generating system. AB - The superoxide-forming NADPH oxidase of human neutrophils was studied in subcellular fractions of unstimulated cells. Purified neutrophils were disrupted by nitrogen cavitation and separated on Percoll density gradients into four fractions: alpha, azurophil granules; beta, mostly specific granules; gamma, plasma membrane, and cytosol. NADPH-dependent O2-. formation by these fractions was quantitated as the rate of superoxide dismutase-inhibitable reduction of ferricytochrome c. In the presence of cytosol, NADPH, and either arachidonic acid (optimum 90 microM) or sodium dodecyl sulfate (optimum 160 microM), 70-75% of the oxidase was in the beta fraction and about 25% was in the gamma fraction. A similar distribution was found for cytochrome b559 and FAD, two putative components of the oxidase. The reaction rates observed with arachidonic acid activation were sufficient to account for 25-75% of the O2-. generated by intact neutrophils. The properties of the beta and gamma enzymes were similar and closely resembled those of the oxidase in intact neutrophils or disrupted prestimulated cells. These included resistance to azide and cyanide, a pH optimum of 7.4, and a preference for NADPH (Km approximately 40-45 microM) rather than NADH (Km approximately 2.5 mM) as the electron donor. The combination of beta and gamma fractions displayed additive activity. The activatable oxidase required Mg2+ but not Ca2+. ATP was required for maximum reaction rates. When beta and gamma membranes were preincubated with cytosol and arachidonic acid in the presence of millimolar Mg2+ and then ultracentrifuged membrane-bound O2-. forming activity was recovered in the pellet and the enzyme required only NADPH (i.e. no cytosol, arachidonic acid, or Mg2+) for expression of activity. These data suggest that cytosol contains a Mg2+-dependent oxidase-activating factor. Molecular sieve chromatography of cytosol indicated a single peak of activity (i.e. ability to activate O2-. generation by beta and/or gamma fraction) eluting with molecules of about 10,000 daltons. PMID- 3031061 TI - The alpha-saturation and terminal events in dolichol biosynthesis. AB - Incubations of 10,000 X g supernatant from rat liver with [3H]mevalonate were performed and the labeling of polyprenols was studied. It was demonstrated that factors like pH, substrate concentration, and presence of detergent not only greatly influence the total incorporation but also the relative distribution of radioactivity among the isoprenologues. The synthesis was shown to be extremely sensitive to Triton X-100. Substrate concentrations of 1 and 100 microM mostly gave polyprenols with 18 and 20 isoprenes, respectively. At a given substrate concentration, pH 6.5 resulted in shorter polyprenols than did pH 7.5. Ozonolytic fragmentation demonstrated that in the initial phase of incubation, polyprenols are elongated by 1 isoprene residue and saturated to give dolichols. No substantial dephosphorylation of polyprenyl phosphates to the free alcohol occurred. The production of dolichol in vitro was shown to utilize NADH for the saturation event. This seemed to occur concomitantly with the synthesis. alpha Saturation of polyprenyl-P could not be achieved with the procedures employed. It is proposed that the synthesis of dolichol and dolichyl-P do not share the same terminal steps; saturation and terminal isoprene condensation occur in cooperation; and substrate concentration and pH influence the terminal enzyme(s) and the nature of the final product in the polyprenol biosynthesis. PMID- 3031062 TI - Receptor-mediated endocytosis in Xenopus oocytes. I. Characterization of the vitellogenin receptor system. AB - We have investigated the vitellogenin (VTG) receptor system in Xenopus oocytes since these cells are specialized for endocytosis. Oocytes have between 0.2 and 3 X 10(11) receptors per 1-mm cell. There is only a single class of receptors of low affinity (1.3 X 10(-6) M at 22 degrees C and 2-4 X 10(-6) M at 0 degree C), but high specificity (less than 5% nonspecific binding at 2 X 10(-6) M). The specific internalization rate of the VTG receptor (around 2 X 10(-3) s-1) is first order, highly variable, and at the upper end of the range of values reported for mammalian cells. The receptor association rate constant (9.6 X 10(2) M-1 s-1) is extremely low although the dissociation rate constant was immeasurable. Calcium is required for VTG binding, and low pH does not dissociate the VTG-receptor complex. Monensin treatment at 100 microM caused the loss of surface receptors with a t1/2 of 3 h and the accumulation of internalized ligand in a "pre-lysosomal" endocytic compartment. Conversely, the recovery of surface VTG receptors that were removed with trypsin occurred with a t1/2 of about 2 h. These observations indicate that oocytes have very large intracellular pools of receptors and that although surface receptors are internalized on the time scale of minutes, the intracellular pool is recycled on the time scale of hours. PMID- 3031064 TI - Interleukin 2 rapidly stimulates synthesis and breakdown of polyphosphoinositides in interleukin 2-dependent, murine T-cell lines. AB - Several T-cell functions are controlled by the regulatory peptide interleukin 2 (IL-2). Binding of IL-2 with specific receptors has been well documented, but the molecular mechanism by which IL-2/IL-2 receptor interaction is transduced is not known. We have found that treatment of IL-2-dependent T-cell lines with IL-2 is followed by a rapid stimulation of inositol phospholipid metabolism, as determined by isotopic methodology employing myo-[1,2-3H]inositol. Increased incorporation of the metabolic precursor into phosphatidylinositol and phosphatidylinositol 4-monophosphate, together with the appearance of radiolabeled phosphatidylinositol 4,5-bisphosphate, occurred within minutes of treatment with IL-2 of factor-dependent CT6 cells. Analysis of labeled water soluble compounds from prelabeled cells indicated a rapid (within 1 min) stimulation of inositol phospholipid hydrolysis following IL-2 treatment. Increased recovery of [3H] inositol phosphates and appearance of [3H]inositol trisphosphate were observed after treatment with IL-2 of CT6 cells, as well as of a second IL-2-dependent cell line, CTB6. These findings suggests that inositol phospholipid-derived metabolites (i.e. diacylglycerol and inositol trisphosphate) may be part of the mechanism by which certain IL-2 signals are transduced. PMID- 3031063 TI - Receptor-mediated endocytosis in Xenopus oocytes. II. Evidence for two novel mechanisms of hormonal regulation. AB - Xenopus oocytes exhibit enhanced rates of vitellogenin (VTG) endocytosis following exposure to insulin in vitro and human chorionic gonadotropin in vivo. We investigated this phenomenon using kinetic and steady state analyses and found that the stimulation of VTG uptake was not due to an increase in surface VTG receptors. Instead, both hormones acted by stimulating the specific internalization rate (ke) of the VTG receptor, although by apparently different mechanisms. The stimulation of ke by insulin was most obvious at low levels of receptor occupancy. Insulin also increased the affinity of the VTG receptor for its ligand. Both hormones increased the rate of fluid phase endocytosis, but the magnitude of stimulation was not sufficient to account for the observed increase in VTG uptake. The steady state binding of VTG was biphasic with respect to increasing ligand concentration, primarily due to a nonlinear receptor internalization rate as a function of occupancy. The nonlinear VTG binding was abolished by incubating the oocytes at 0 degree C. Our data are consistent with a model in which there is a cell surface regulatory component that facilitates the internalization of the occupied VTG receptor. Although both human chorionic gonadotropin and insulin stimulate VTG uptake by increasing the net rate of endocytosis, insulin also increases the association of the occupied VTG receptor with this regulatory component. PMID- 3031066 TI - Complementation of mutations and nucleotide sequence of FAS1 gene encoding beta subunit of yeast fatty acid synthase. AB - The yeast fatty acid synthase is a complex (alpha 6 beta 6) of two multifunctional proteins alpha and beta. The alpha subunit (Mr 212,000) contains two of the seven enzymatic activities required for the synthesis of fatty acids and the site for attachment of the prosthetic group 4'-phosphopantetheine. The beta subunit (Mr 203,000) contains the remaining five activities. Cloning of the genes encoding the alpha and beta proteins has been reported (Kuziora, M. A., Chalmers, J. H., Jr., Hitzeman, R. A., Douglas, M. G., and Wakil, S. J. (1983) J. Biol. Chem. 258, 11648-11653). In the present study it is shown that two of the clones containing the beta subunit gene, YEpFAS1 and YEp33F1, are not identical. The clone YEp33F1 contains the gene that codes for the entire beta subunit while YEpFAS1 is missing approximately half of the gene at the 3' end. Despite this loss, YEpFAS1 is still able to complement a fas1 mutation at the enoyl reductase domain. This complementation does not occur by recombination, rather a small mRNA is produced in cells transformed with YEpFAS1 and is translated into a protein of molecular weight of approximately 125,000 which is immunologically reactive with yeast fatty acid synthase antibodies. The data suggest that this truncated beta subunit interacts with the mutant alpha 6 beta 6 complex to restore fatty acid synthesis to the cell. The nucleotide sequence of the FAS1 gene cloned in YEp33F1 DNA, which encodes the beta subunit of fatty acid synthase, was determined. The coding region consists of 5940 base pairs (bp) and could encode a protein of 1980 amino acids with a calculated molecular weight of 220,077. A major transcriptional start point was mapped to a position of about 330 bp upstream from the first ATG codon. The termination of transcription was mapped at about 300 bp downstream from the first TGA stop codon. The sequence of the beta subunit protein does not appear to be similar to any other sequenced protein. The sites of the active seryl groups for the acetyltransacylase and malonyl/palmitoyl transacylase were identified from known amino acid sequences to be residues 274 and 1808, respectively. Putative binding sites for FMN and NADPH were suggested based on similarities with amino acid sequences of known flavin and pyridine nucleotide enzymes, respectively. PMID- 3031065 TI - Alpha 1-adrenergic receptor-mediated phosphoinositide hydrolysis and prostaglandin E2 formation in Madin-Darby canine kidney cells. Possible parallel activation of phospholipase C and phospholipase A2. AB - alpha 1-Adrenergic receptors mediate two effects on phospholipid metabolism in Madin-Darby canine kidney (MDCK-D1) cells: hydrolysis of phosphoinositides and arachidonic acid release with generation of prostaglandin E2 (PGE2). The similarity in concentration dependence for the agonist (-)-epinephrine in eliciting these two responses implies that they are mediated by a single population of alpha 1-adrenergic receptors. However, we find that the kinetics of the two responses are quite different, PGE2 production occurring more rapidly and transiently than the hydrolysis of phosphoinositides. The antibiotic neomycin selectively decreases alpha 1-receptor-mediated phosphatidylinositol 4,5 bisphosphate hydrolysis without decreasing alpha 1-receptor-mediated arachidonic acid release and PGE2 generation. In addition, receptor-mediated inositol trisphosphate formation is independent of extracellular calcium, whereas release of labeled arachidonic acid is largely calcium-dependent. Moreover, based on studies obtained with labeled arachidonic acid, receptor-mediated generation of arachidonic acid cannot be accounted for by breakdown of phosphatidylinositol monophosphate, phosphatidylinositol bisphosphate, or phosphatidic acid. Further studies indicate that epinephrine produces changes in formation or turnover of several classes of membrane phospholipids in MDCK cells. We conclude that alpha 1 adrenergic receptors in MDCK cells appear to regulate phospholipid metabolism by the parallel activation of phospholipase C and phospholipase A2. This parallel activation of phospholipases contrasts with models described in other systems which imply sequential activation of phospholipase C and diacylglycerol lipase or phospholipase A2. PMID- 3031068 TI - Interaction between the DNA polymerase and single-stranded DNA-binding protein (infected cell protein 8) of herpes simplex virus 1. AB - The herpes virus-encoded DNA replication protein, infected cell protein 8 (ICP8), binds specifically to single-stranded DNA with a stoichiometry of one ICP8 molecule/12 nucleotides. In the absence of single-stranded DNA, it assembles into long filamentous structures. Binding of ICP8 inhibits DNA synthesis by the herpes induced DNA polymerase on singly primed single-stranded DNA circles. In contrast, ICP8 greatly stimulates replication of circular duplex DNA by the polymerase. Stimulation occurs only in the presence of a nuclear extract from herpes-infected cells. Appearance of the stimulatory activity in nuclear extracts coincides closely with the time of appearance of herpes-induced DNA replication proteins including ICP8 and DNA polymerase. A viral factor(s) may therefore be required to mediate ICP8 function in DNA replication. PMID- 3031067 TI - Processive replication of single-stranded DNA templates by the herpes simplex virus-induced DNA polymerase. AB - The DNA polymerase encoded by herpes simplex virus 1 consists of a single polypeptide of Mr 136,000 that has both DNA polymerase and 3'----5' exonuclease activities; it lacks a 5'----3' exonuclease. The herpes polymerase is exceptionally slow in extending a synthetic DNA primer annealed to circular single-stranded DNA (turnover number approximately 0.25 nucleotide). Nevertheless, it is highly processive because of its extremely tight binding to a primer terminus (Kd less than 1 nM). The single-stranded DNA-binding protein from Escherichia coli greatly stimulates the rate (turnover number approximately 4.5 nucleotides) by facilitating the efficient binding to and extension of the DNA primers. Synchronous replication by the polymerase of primed single-stranded DNA circles coated with the single-stranded DNA-binding protein proceeds to the last nucleotide of available 5.4-kilobase template without dissociation, despite the 20-30 min required to replicate the circle. Upon completion of synthesis, the polymerase is slow in cycling to other primed single-stranded DNA circles. ATP (or dATP) is not required to initiate or sustain highly processive synthesis. The 3'----5' exonuclease associated with the herpes DNA polymerase binds a 3' terminus tightly (Km less than 50 nM) and is as sensitive as the polymerase activity to inhibition by phosphonoacetic acid (Ki approximately 4 microM), suggesting close communication between the polymerase and exonuclease sites. PMID- 3031069 TI - Cell-free fatty acylation of microsomal integrated and detergent-solubilized glycoprotein of vesicular stomatitis virus. AB - An enzymatic activity associated with intracellular membrane fractions of Merwin plasma cell tumor II, baby hamster kidney, and chicken embryo fibroblast cells and bovine kidney has been characterized which covalently links fatty acids onto the G protein of vesicular stomatitis virus. Exogenous G protein extracted from native vesicular stomatitis virus particles can be acylated in vitro only after it has been previously deacylated. The fatty acids transferred in vitro are sensitive to treatment with hydroxylamine, indicating an ester linkage. Cell-free acyl transfer was also observed with endogenous G protein present in membrane fractions prepared from vesicular stomatitis virus-infected cells. In this case, the fatty acids become linked to a G protein species (G1) which is not terminally glycosylated and therefore has not entered the trans-Golgi compartment. The same G protein species also becomes acylated in infected cells during short pulses with radioactive palmitic acid. Acylation of the G protein in vitro with free palmitic or myristic acid is energy-dependent, and the addition of ATP is specifically required. Other nucleoside triphosphates cannot substitute for ATP in the activation of free acyl chains. Alternatively, activated fatty acids linked in a high energy thioester bond to coenzyme A, e.g. [14C] palmitoyl-CoA, are suitable lipid donors in the in vitro acylation reactions. Palmitic acid transfer onto G protein shows the typical characteristics of an enzyme-catalyzed reaction. PMID- 3031070 TI - Permeability of the peroxisomal membrane to cofactors of beta-oxidation. Evidence for the presence of a pore-forming protein. AB - Peroxisomes were purified from livers of clofibrate-treated rats. Permeability measurements on the isolated organelles revealed that peroxisomes are permeable to small solutes, including sucrose and the cofactors for fatty acid oxidation NAD+, CoA, ATP, and carnitine. The intraperoxisomal distribution volume was equal for all solutes. Peroxisomal solute uptake was rapid, not saturable and not visibly influenced by temperature. NAD+ and carnitine uptake in the solute accessible volume was not diminished by a variety of analogs and inhibitors. Subfractionation of peroxisomes and reconstitution of the subfractions into liposomes preloaded with solutes made the liposomes reconstituted with the integral membrane protein fraction, but not those reconstituted with the other subperoxisomal protein fractions, permeable to the same solutes that entered intact peroxisomes. Solute leakage from the preloaded liposomes was rapid and not visibly influenced by temperature. Leakage activity was destroyed by heat treatment of the integral membrane protein fraction and was not present in lipid extracts of the membrane. Separation of the integral membrane proteins on sucrose density gradients and reconstitution of the gradient fractions into liposomes indicated that the leakage activity was caused by a polypeptide of rather low molecular weight. The gradient distribution of leakage activity corresponded most closely to the presence of a 22- and a 28-kDa polypeptide. Our experiments indicate that the nonspecific permeability of the peroxisomal membrane to small solutes is based on the presence in the membrane of a nonselective pore-forming protein. PMID- 3031071 TI - Antipeptide antibodies directed against cytoplasmic rhodopsin sequences recognize the beta-adrenergic receptor. AB - Antibodies were made against synthetic peptides that correspond to cytoplasmic domains of rhodopsin, the photopigment protein of the retinal rod. These antipeptide antibodies recognized rhodopsin as detected by immunoblot analysis. Antibodies directed against the cytoplasmic loop between transmembrane domains 1 and 2, as well as those directed against the serine/threonine-rich region of the COOH terminus of bovine rhodopsin, also recognized purified beta-adrenergic receptor isolated from mouse S49 lymphoma cells. In addition, antibodies raised against membrane-associated rhodopsin recognized the beta-adrenergic receptor. Both the antipeptide and anti-rhodopsin antibodies were able to detect a 65-kDa protein band corresponding to the molecular weight of the beta-adrenergic receptor in membranes derived from human placenta, rat adipocytes, and S49 mouse lymphoma cells. Putative recognition sites for the rhodopsin antibodies on the beta-adrenergic receptor are identified, and the significance of the homology between the two proteins is discussed. PMID- 3031072 TI - A study of the mechanism of glucagon-induced protein phosphorylation in isolated rat hepatocytes using (Sp)-cAMPS and (Rp)-cAMPS, the stimulatory and inhibitory diastereomers of adenosine cyclic 3',5'-phosphorothioate. AB - Maximal doses of glucagon increase the phosphorylation state of 12 cytosolic proteins in isolated hepatocytes from fasted rats (Garrison, J. C., and Wagner, J. D. (1982) J. Biol. Chem. 257, 13135-13143). Incubation of hepatocytes with lower concentrations of glucagon indicates that a hierarchy of substrates exists with the concentration of glucagon required for half-maximal increases in phosphorylation varying 5-15-fold. The proteins whose phosphorylation state is most sensitive to low concentrations of glucagon are pyruvate kinase and 6 phosphofructo-2-kinase/fructose-2,6-bisphosphatase, both of which play key roles in the regulation of gluconeogenesis. Treatment of hepatocytes with (Sp)-cAMPS, the stimulatory diastereomer of adenosine cyclic 3',5'-phosphorothioate, mimics the response seen with glucagon. When hepatocytes are pretreated with the cAMP antagonist, (Rp)-cAMPS, the phosphorylation response is abolished at low concentrations of glucagon, and the dose of glucagon required for half-maximal stimulation of phosphorylation is increased 5-10-fold. The (Sp)-cAMPS-stimulated increases in phosphorylation state are also blunted by (Rp)-cAMPS. These results provide direct pharmacological evidence for the activation of the cAMP-dependent protein kinase in response to glucagon in the intact cell. Although low doses of glucagon appear to stimulate protein phosphorylation via the cAMP-dependent protein kinase, high doses of glucagon also cause a small increase in the concentration of free intracellular Ca2+ in hepatocytes. The glucagon-stimulated increases in the level of Ca2+ can be mimicked by (Sp)-cAMPS and inhibited by pretreatment with (Rp)-cAMPS. These results suggest that glucagon can elevate intracellular Ca2+ via cAMP and the cAMP-dependent protein kinase. PMID- 3031073 TI - Molecular cloning of a cDNA for a human ADP/ATP carrier which is growth regulated. AB - We have identified in a human cDNA library a clone (hp2F1) whose cognate RNA is growth-regulated. The insert has been sequenced and the nucleotide sequence shows a strong homology to the nucleotide sequences of the ADP/ATP carrier cDNA and gene, respectively, isolated from Neurospora crassa and Saccharomyces cerevisiae. The putative amino acid sequence of hp2F1 shows an 87% homology to the amino acid sequence of the ADP/ATP carrier from beef heart mitochondria. We conclude that the insert of hp2F1 contains the full coding sequence of a human ADP/ATP carrier. The steady-state RNA levels of the ADP/ATP carrier are growth-regulated. They increase when quiescent cells are stimulated by serum, platelet-derived growth factor, or epidermal growth factor, but not by platelet-poor plasma or insulin. RNA levels of the ADP/ATP carrier decrease instead when growing HL-60 cells are induced to differentiate by either phorbol esters or retinoic acid. PMID- 3031074 TI - Glucose-1-phosphotransferase and N-acetylglucosamine-1-phosphotransferase have distinct acceptor specificities. AB - UDP-glucose:glycoprotein glucose-1-phosphotransferase (Glc-phosphotransferase) catalyzes the transfer of alpha Glc-1-P from UDP-Glc to endoglycosidase H sensitive oligosaccharides on acceptor glycoproteins. We have previously demonstrated that Glc-phosphotransferase was specific for UDP-Glc as its nucleotide sugar substrate and thus appeared to be distinct from UDP-N acetylglucosamine:glycoprotein N-acetylglucosamine-1-phosphotransferase (GlcNAc phosphotransferase), an enzyme specific for lysosomally destined acceptor glycoproteins. Here, sodium dodecyl sulfate-polyacrylamide gel electrophoresis autoradiographs of endogenous acceptor glycoproteins in embryonic chick neural retina homogenates labeled by the presence of [beta-32P]UDP-Glc were shown to be distinct from those labeled by [beta-32P]UDP-GlcNAc, indicating that the two enzymatic activities recognize different populations of endogenous glycoproteins. To further probe the acceptor specificities of these enzymes, three glycoproteins known to be exogenous acceptors for GlcNAc-phosphotransferase were included in assays for Glc-phosphotransferase from retinal homogenates. Cathepsin D and beta N-acetylhexosaminidase had no significant effects on phosphoglucose incorporation. Uteroferrin, an acid phosphatase, had a pronounced inhibitory effect on incorporation from UDP-Glc, and subsequent experiments suggested that phosphorylation of the Glc-phosphotransferase or another protein may be necessary for maximal activity to be seen. Also, I-cells, which have previously been shown to possess no GlcNAc-phosphotransferase activity, and control human fibroblasts were assayed for both Glc-phosphotransferase and GlcNAc-phosphotransferase. GlcNAc-phosphotransferase activity was observed only in control cells, whereas Glc-phosphotransferase was observed in both I-cells and controls at similar specific activities. PMID- 3031075 TI - Structure, assembly, and secretion of octameric invertase. AB - Yeast invertase forms a homo-octamer of core glycosylated subunits during assembly in the lumen of the endoplasmic reticulum. This form has been purified from mutant cells (sec18) in which transport of secreted proteins from the endoplasmic reticulum is blocked. No heterologous protein subunits are found in the purified material. Analysis of invertase derived from wild type cells or from mutant cells blocked at subsequent stages in secretion demonstrates that invertase remains a homo-octamer throughout the pathway even though the extent of subunit glycosylation increases. Purified octameric invertase is dissociated into dimer units that reassociate in the presence of polyethylene glycol. Negatively stained preparations show the dissociated enzyme as individual spheres, whereas octameric invertase appears as four associated spheres. Assembly of the octamer in vitro and in vivo is facilitated by the presence of N-linked carbohydrate. Selective release of dimeric glycosylated invertase from intact yeast cells suggests that oligomerization helps retain the enzyme in the periplasmic space. PMID- 3031076 TI - The potency determination of human varicella-zoster immunoglobulin by enzyme linked immunosorbent assay, complement-fixation test and indirect fluorescent antibody tests. AB - Traditionally, plasma for the production of the human varicella-zoster immunoglobulin (VZIG) has been selected on the basis of the complement-fixing antibody (CFA) titre. Since immune individuals may lack CFA to varicella-zoster virus (VZV), non-CFA may be of importance in protection. In a search for a simple and reliable method for potency determination, 24 VZIG preparations were quantified by enzyme-linked immunosorbent assay (ELISA), the complement-fixation test (CFT), the indirect fluorescent antibody test to acetone-fixed (IF) and viable (FAMA) VZV-infected cells, respectively. The antibody titres obtained by the various methods were compared. Arranged in order of decreasing agreement, the correlation coefficients (r) of the regression equations between the variables were 0.62 for CFT and FAMA, 0.50 for CFT and ELISA and 0.26 for CFT and IF in a log2 plot. There was complete agreement between the titres obtained by the commercially available Enzygnost Varicella/Zoster kits (Behring Institute, Marburg, F.R. Germany) and the ELISA microtitre plates produced at our institute (r = 1). The regression equation lines for ELISA/CFT and FAMA/CFT titres tended to be parallel to each other, while the line for IF/CFT titres had a less steep slope. Similar titration curves were obtained for VZIGs fractionated by two different methods. Furthermore, the titration curves of serum pools from varicella and zoster convalescents, respectively, had a similar shape below delta OD = 0.4. Generally, a steeper slope was observed above delta OD = 0.4. As antibody detectable by ELISA seems to correlate with protection and the method is sensitive, specific, reproduceable, simple to carry out and easily automated, it may be suitable for the potency determination of VZIGs. PMID- 3031077 TI - Synthetic peptide vaccines against foot-and-mouth disease. I. Duration of the immune response and priming in guinea-pigs, rabbits and mice. AB - The immunogenicity of two aphthovirus-specific synthetic peptides was investigated. One peptide copied the sequence of amino acids 141 to 160 from the capsid VP1 of the aphthovirus strains O1 BFS 1860 and O1 Kaufbeuren (O peptide), the other copied the equivalent sequence from aphthovirus strain A24 Cruzeiro (A peptide). Peptide coupled to keyhole limpet haemocyanin (KLH) stimulated a long lasting immune response in guinea-pigs and rabbits. Significant levels of antibody were detectable at least one year after vaccination, although the reactivity of the antibody depended on the species and the peptide used. In some circumstances the peptides were able to prime the immune system such that a subsequent dose of peptide boosted antibody production. This effect, also, was dependent on the species of experimental animal and on the peptide used, an observation which has important implications for the use of such peptides as vaccines. PMID- 3031078 TI - Synthetic peptide vaccines against foot-and-mouth disease. II. Comparison of the response of guinea-pigs, rabbits and mice to various formulations. AB - The immune response of guinea-pigs to vaccination with either of two aphthovirus specific synthetic peptides was investigated. One peptide copied the sequence of amino acids 141 to 160 from the VP1 of aphthovirus strains O1BFS 1860 and O1 Kaufbeuren (O peptide), the other copied the equivalent sequence from aphthovirus strain A24 Cruzeiro (A peptide). The immune response was enhanced when the peptides were conjugated to a carried protein, but the choice of carrier protein and cross-linking agent was not critical in obtaining enhancement. The response was greatest when the peptide or peptide-carrier conjugate was adjuvanted in Freund's complete adjuvant (FCA). The O peptide was poorly immunogenic and did not confer protection against challenge with infectious virus, whereas the A peptide had good immunogenicity and did confer protection. This reflected the relative immunogenicity of the parent viruses. Rabbits, three strains of guinea pigs and seven strains of mice were vaccinated with the O peptide conjugated to keyhole limpet haemocyanin (KLH). Considerable variation was observed between the responses of each strain and it is proposed that this relates to their repertoire of immune response genes. PMID- 3031080 TI - Metabolic denitrosation of diphenylnitrosamine: a possible bioactivation pathway. AB - Nitrosodiphenylamine was tested for induction of DNA single strand breaks in rat hepatocytes and Chinese hamster V 79 cells with the alkaline filter elution assay. While in rat hepatocytes DNA damage was observed, negative results were obtained in V 79 cells. In view of the metabolic capacity of hepatocytes and the chemical structure of nitrosodiphenylamine it seems likely that cytochrome P-450 dependent, reductive denitrosation might be necessary for exerting this effect. Therefore the metabolism of nitrosodiphenylamine was investigated in phenobarbital-induced mouse liver microsomes and some of the metabolites were also tested. One metabolite was identified as diphenylamine whereas the others were identified as a ring-hydroxylated derivative of diphenylamine and its corresponding quinoneimine. Diphenylhydroxylamine which was not detected in the microsomes as a metabolite produced a significant amount of DNA single strand breaks in V 79 cells. When diphenylhydroxylamine was incubated with microsomes electron spin resonance spectrum was observed which indicated the formation of the diphenylnitroxide radical. This radical seems to be mediated by auto oxidation rather than by enzymatic catalysis. Whether diphenylhydroxylamine might be responsible for the observed genetoxic effects of nitrosodiphenylamine assumed to be produced via active oxygen species is discussed. PMID- 3031079 TI - Targeting of neoglycoprotein-drug conjugates to cultured human embryonal carcinoma cells. AB - Fluorescent neoglycoproteins were used to screen for the presence and sugar specificities of cell surface lectins in two human embryonal carcinoma cell lines. Efficient labeling correlated with extent of lectin-mediated uptake of neoglycoproteins, as measured by inhibition of DNA synthesis by drug neoglycoprotein conjugates. These conjugates contain covalently linked carbohydrate moieties on the carriers to render them accessible to the membrane lectins, most effectively galactosides and alpha-glucosides. They furthermore contain chemically linked cytotoxic drugs (etoposide, cis diamminedichloroplatinum II and methotrexate) which are intracellularly released after lysosomal breakdown of the carrier, as indicated by the effect of leupeptin. Sugars can confer a greater than 10-fold increase in cytotoxic capacity to the nonglycosylated carrier-drug conjugate, nearly reaching the level of toxicity of the freely diffusible drug. Two different neoglycoproteins, reacting with independently targeted membrane lectins, were shown to be useful in a model for combination chemotherapy. These results therefore suggest potential usefulness of custom-made glycosylated carriers in the targeting of therapeutic agents to human embryonal carcinoma cells. PMID- 3031081 TI - Posttranslational modifications of the cytochrome P-450 monooxygenase system. AB - Two forms of enzymatic posttranslational modifications of the monooxygenase system are described: modification by phosphatase and modification by protein kinase. Phosphatase treatment of microsomes isolated from phenobarbital pretreated rabbits and rats caused a marked decrease of monooxygenase activity which was paralleled by a comparable decrease of NADPH-cytochrome P-450 reductase activity while the second essential component of the system, cytochrome P-450, remained unaltered. Thus phosphatase attacks monooxygenase via reductase. Protein kinases showed the opposite preference; while cytochrome P-450 was phosphorylated, NADPH-cytochrome P-450 reductase was not. Thus the kinase affects monooxygenase via cytochrome P-450. The phosphorylation of cytochrome P-450 turned out to be a specific reaction observed only with certain cytochrome P-450 isoenzymes and certain protein kinases. PMID- 3031082 TI - Accumulation of adrenocorticotropin secretory granules in the midbody of telophase AtT20 cells: evidence that secretory granules move anterogradely along microtubules. AB - During the cell cycle the distribution of the ACTH-containing secretory granules in AtT20 cells, as revealed by immunofluorescence labeling and electron microscopy of thin sections, undergoes a cycle of changes. In interphase cells the granules are concentrated in the Golgi region, where they form, and also at the tips of projections from the cells, where they accumulate. These projections contain many microtubules extending to their tips. During metaphase and anaphase the granules are randomly distributed in the cytoplasm of the rounded-up mitotic cells. On entry into telophase there is a rapid and striking redistribution of the granules, which accumulate in large numbers in the midbody as it develops during cytokinesis. This accumulation of secretory granules in the midbody is dependent upon the presence of microtubules. The changing pattern of distribution of the secretory granules during the cell cycle fulfills the predictions of a model envisaging first that secretory granules associate with and move along interphase microtubules in a net anterograde direction away from the centrioles, and secondly that they do not associate with microtubules of the mitotic spindle during metaphase and anaphase. PMID- 3031083 TI - Urokinase-type plasminogen activator: proenzyme, receptor, and inhibitors. PMID- 3031084 TI - Isolation and characterization of cDNA clones for rat ribophorin I: complete coding sequence and in vitro synthesis and insertion of the encoded product into endoplasmic reticulum membranes. AB - Ribophorins I and II are two transmembrane glycoproteins that are characteristic of the rough endoplasmic reticulum and are thought to be part of the apparatus that affects the co-translational translocation of polypeptides synthesized on membrane-bound polysomes. A ribophorin I cDNA clone containing a 0.6-kb insert was isolated from a rat liver lambda gtll cDNA library by immunoscreening with specific antibodies. This cDNA was used to isolate a clone (2.3 kb) from a rat brain lambda gtll cDNA library that contains the entire ribophorin I coding sequence. SP6 RNA transcripts of the insert in this clone directed the in vitro synthesis of a polypeptide of the expected size that was immunoprecipitated with anti-ribophorin I antibodies. When synthesized in the presence of microsomes, this polypeptide, like the translation product of the natural ribophorin I mRNA, underwent membrane insertion, signal cleavage, and co-translational glycosylation. The complete amino acid sequence of the polypeptide encoded in the cDNA insert was derived from the nucleotide sequence and found to contain a segment that corresponds to a partial amino terminal sequence of ribophorin I that was obtained by Edman degradation. This confirmed the identity of the cDNA clone and established that ribophorin I contains 583 amino acids and is synthesized with a cleavable amino terminal insertion signal of 22 residues. Analysis of the amino acid sequence of ribophorin I suggested that the polypeptide has a simple transmembrane disposition with a rather hydrophilic carboxy terminal segment of 150 amino acids exposed on the cytoplasmic face of the membrane, and a luminal domain of 414 amino acids containing three potential N-glycosylation sites. Hybridization measurements using the cloned cDNA as a probe showed that ribophorin I mRNA levels increase fourfold 15 h after partial hepatectomy, in confirmation of measurements made by in vitro translation of liver mRNA. Southern blot analysis of rat genomic DNA suggests that there is a single copy of the ribophorin I gene in the haploid rat genome. PMID- 3031085 TI - Rapid analytical and preparative isolation of functional endosomes by free flow electrophoresis. AB - Endosomes are prelysosomal organelles that serve as an intracellular site for the sorting, distribution, and processing of receptors, ligands, fluid phase components, and membrane proteins internalized by endocytosis. Whereas the overall functions of endosomes are increasingly understood, little is known about endosome structure, composition, or biogenesis. In this paper, we describe a rapid procedure that permits analytical and preparative isolation of endosomes from a variety of tissue culture cells. The procedure relies on a combination of density gradient centrifugation and free flow electrophoresis. It yields a fraction of highly purified, functionally intact organelles. As markers for endosomes in Chinese hamster ovary cells, we used endocytosed horseradish peroxidase, FITC-conjugated dextran, and [35S]methionine-labeled Semliki Forest virus. Total postnuclear supernatants, crude microsomal pellets, or partially purified Golgi fractions were subjected to free flow electrophoresis. Endosomes and lysosomes migrated together as a single anodally deflected peak separated from most other organelles (plasma membrane, mitochondria, endoplasmic reticulum, and Golgi). The endosomes and lysosomes were then resolved by centrifugation in Percoll density gradients. Endosomes prepared in this way were enriched up to 70 fold relative to the initial homogenate and were still capable of ATP-dependent acidification. By electron microscopy, the isolated organelles were found to consist of electron lucent vacuoles and tubules, many of which could be shown to contain an endocytic tracer (e.g., horseradish peroxidase). SDS PAGE analysis of integral and peripheral membrane proteins (separated from each other by condensation in Triton X-114) revealed a unique and restricted subset of proteins when compared with lysosomes, the unshifted free flow electrophoresis peak, and total cell protein. Altogether, the purification procedure takes 5-6 h and yields amounts of endosomes (150-200 micrograms protein) sufficient for biochemical, immunological, and functional analysis. PMID- 3031086 TI - gamma-Aminobutyric acid-containing terminals can be apposed to glycine receptors at central synapses. AB - The distributions of terminals containing gamma-aminobutyric acid (GABA) and of endings apposed to glycine receptors were investigated cytochemically in the ventral horn of the rat spinal cord. For this purpose, a polyclonal antibody raised to recognize glutamic acid decarboxylase (GAD), a synthetic enzyme for GABA, and three monoclonal antibodies (mAb's) directed against the glycine receptor were used. Double immunofluorescence showed that, surprisingly, GAD positive terminals are closely associated in this system with glycine receptors at all the investigated cells, most of which were spinal motoneurons. Furthermore, double labeling was performed with immunoenzymatic recognition of GAD and indirect marking of mAb's with colloidal gold. With this combined approach, it was found, at the electron microscopic level, that all GAD-positive terminals are in direct apposition with glycine receptors while, on the other hand, not all glycine receptors are in front of GABA-containing boutons. This result is not due to a cross-reactivity of mAb's with GABA receptors as shown by using as a control synapses known to use GABA as a neurotransmitter in the cerebellar cortex. Indeed, no glycine receptor immunoreactivity was detected on Purkinje cells facing basket axon terminals. However, Purkinje neurons can express glycine receptor immunoreactivity at other synaptic contacts. Assuming that the presence of postsynaptic receptors for glycine indicates that this amino acid is used for neurotransmission at a given synapse, our results strongly support the notion that GABA and glycine, two classical inhibitory transmitters, coexist at some central connections. However, such is not always the case; in the cerebellum, Golgi terminals impinging on the dendrites of granule cells are either GAD-positive or face glycine receptors, in a well-segregated manner. PMID- 3031087 TI - Further analysis of spontaneous membrane potential activity and the hyperpolarizing response to parathyroid hormone in osteoblastlike cells. AB - Whole cell voltage clamp measurements using the patch technique on well-attached and well-spread cells of an osteoblastlike line (ROS 17/2.8) show the same spontaneous membrane potential activity as measurements with inserted microelectrodes. Furthermore, membrane potential measurements during the first 80 milliseconds (ms) following microelectrode penetration of the cell membrane usually show no decay. There is also good agreement between values of cell membrane resistance obtained by the microelectrode technique, the whole cell patch clamp technique, and the single channel patch clamp technique. These results indicate that our microelectrode measurements are not dominated by leak induced artifacts, and that the spontaneous membrane potential activity is not induced by Ca2+ leakage around the microelectrode. The spontaneous membrane potential activity is eliminated in the presence of the Ca2+ ionophore A23187, also in serum-free medium, and by K+ and Ca2+ channel blockers, but it is not affected by the hyperpolarizing responses to parathyroid hormone (PTH) and dibutyryl cAMP, which persist under all of these conditions. These results support the hypothesis that the spontaneous membrane potential activity is related to repeated fluctuations of internal [Ca2+] and that such fluctuations result from a feedback loop involving Ca2+ channels or Ca2+ pumps in the cell membrane. PMID- 3031088 TI - Heat stress stimulates inositol trisphosphate release and phosphorylation of phosphoinositides in CHO and Balb C 3T3 cells. AB - Exposure of eukaryotic cells to elevated temperature leads to profound switches in cell metabolism and gene expression which may be involved in cellular homeostatic mechanisms. We have investigated the effect of heat shock (45 degrees C) on the metabolism of the phosphoinositides, a class of phospholipids involved in the function of Ca2+ -linked membrane receptors. Heat shock led to stimulation of phosphoinositide turnover in HA1-CHO and Balb C 3T3 cells, resulting in the rapid accumulation of inositol trisphosphate (IP3). Mitogenic and alpha 1 adrenergic stimulation, with serum or phenylephrine, led to similar increases in IP3. Heat shock also caused rapid increase in phosphorylation of polyphosphoinositides (PPI). Prolonged exposure to heat greater than 15 min at 45 degrees C led to progressive cellular toxicity which was associated with depletion of PPI. This decline in PPI concentration appeared to result from inhibition of PPI resynthesis. In this respect, heat may resemble some other types of cellular stresses in stimulating membrane phospholipases to deplete classes of membrane phospholipids. The induction of PPI turnover may, therefore, be involved in both pleiotropic responses to brief heat shock and toxicity resulting from prolonged thermal stress. PMID- 3031089 TI - Modulation by the src oncogene of the effect of inhibitory diffusible factor IDF45. AB - Density-dependent inhibition of growth has been assumed to be under the control of inhibitory molecules diffusing from dense cell cultures. Growth inhibitory factors have been fractionated or purified from medium conditioned by different cell types. In the present work, it was shown that IDF45 (inhibitory factor diffusing from 3T3 cells) decreased DNA synthesis in chick embryo fibroblasts (CEF) and was an inhibitor of CEF growth; this inhibition was reversible. Since similitudes between oncogene products and growth factors have been observed, it was of interest to compare the inhibitory effect of IDF45 upon the stimulation of DNA synthesis induced either by serum or by pp60-src. CEF infected by Ny68 virus (a mutant of Rous sarcoma virus ts for the expression of transformation) were density-inhibited at 41 degrees C, but were stimulated at this temperature by addition of 1% serum. This stimulation was 94% inhibited by IDF45. The same Ny68 infected cells could also be stimulated by transfer to 37 degrees C, the permissive temperature (in the absence of serum). The stimulation of DNA synthesis by src expression was poorly inhibited by IDF45. From our results, it appears that oncogene expression in CEF induces a loss in their sensitivity to IDF45. This would explain why transformed cells escape DDI of growth. PMID- 3031090 TI - Isolation and characterization of a cloned growth factor dependent macrophage cell line, BAC1.2F5. AB - The SV40 transformed murine macrophage cell line, BAC1, proliferates in response to the colony stimulating factor, CSF-1 (Schwarzbaum et al., J. Immunol., 132:1158, 1984). In order to obtain a cell line suitable for biochemical and genetic studies of CSF-1 signal transduction, clones of BAC1 were established. Clones ranged from being completely autonomous to being completely dependent on CSF-1 for growth. Cells of one clone (2F5), which proliferated in response to either CSF-1 or granulocyte-macrophage CSF (GM-CSF) were characterized in detail. The kinetics of receptor-mediated internalization and intracellular destruction of CSF-1 were comparable to the kinetics observed with peritoneal exudate macrophages. CSF-1 was shown to regulate cell spreading, cell survival, protein degradation, and the duration of the G1 and S phases of the cell cycle. The 2F5 clone therefore exhibits a number of CSF-1 stimulated responses and is being used for genetic and biochemical studies of CSF-1 action. PMID- 3031091 TI - Exogenous norepinephrine constricts cerebral arterioles via alpha 2-adrenoceptors in newborn pigs. AB - The purpose of this study was to determine whether exogenous norepinephrine mediates cerebrovascular constriction via alpha 1- or alpha 2-adrenoceptors in anesthetized neonatal pigs. Diameters of pial arterioles in anesthetized piglets, 1--6 days old, were investigated using a "closed" cranial window. We examined constrictor effects of norepinephrine on pial arterioles in the absence and presence of relatively selective alpha 1-(prazosin) and alpha 2-(yohimbine) adrenoceptor antagonists (1 mg/kg i.v.). Yohimbine and prazosin inhibited pial arteriolar constriction induced by topical application of clonidine and phenylephrine (10(-6) and 10(-4) M, respectively), and yohimbine did not affect the response to topical phenylephrine. In one group diameter was 188 +/- 13 (mean +/- SEM) micron during control and 146 +/- 12 micron during 10(-5) M norepinephrine (22 +/- 5% constriction). Following yohimbine the same vessels did not constrict significantly. In another group 10(-5) M norepinephrine constricted arterioles by 22 +/- 5%, and this response was unaffected by prazosin (24 +/- 5% constriction). We conclude that pial arterioles are responsive to both alpha 1- and alpha 2-adrenoceptor agonists, that intravenous administration of prazosin and yohimbine results in these drugs crossing the blood-brain barrier and inhibiting constrictor effects of agonists, and that norepinephrine constricts pial arterioles predominantly via alpha 2-adrenoceptors. PMID- 3031092 TI - [Spontaneous rupture of an hepatocarcinoma on a cirrhotic liver]. PMID- 3031093 TI - Neurologic complications of intravenous drug abuse. PMID- 3031094 TI - Leg paralysis in man with oat cell carcinoma. PMID- 3031095 TI - High-performance immobilized-metal affinity chromatography of proteins on iminodiacetic acid silica-based bonded phases. AB - High-performance separations of proteins, based on immobilized-metal affinity chromatography (HPIMAC), are described. The stationary phase consisted of iminodiacetic acid (IDA) chelate groups, bonded to small particle, wide pore silica gel by means of a polyether hydrophilic leash. After loading the column with metal, retention of proteins was achieved by protein-metal complexation at high concentrations of sodium sulfate. Elution was accomplished by addition of competitive complexing ligands, such as ammonia at constant pH, and/or by a decreasing pH gradient of a specially designed buffer system to maintain buffer capacity constant throughout the gradient. Selective separations, based on differences in the number of histidine residues present on the surface of the proteins, are described. The application of HPIMAC in the separation and purification of structurally similar proteins is presented. The potential application of IDA columns in three chromatographic modes (HPIMA, hydrophobic interaction, and cation exchange) is also described. PMID- 3031096 TI - Quantitative analytical aspects of reversed-phase liquid chromatography with slurry-packed capillary columns. AB - Conditions for reproducible packing of fused-silica liquid chromatography capillary columns were demonstrated. The plate height vs. velocity curves were determined for successively prepared columns. In addition, the values of separation impedance were calculated. Several liquid chromatography systems were evaluated in conjunction with the slurry-packed capillaries for the retention and peak area reproducibility. PMID- 3031097 TI - High-performance liquid chromatographic separation of biological macromolecules on new silica-based ion exchangers. AB - The fundamental characteristics of a new series of ion-exchange columns, specifically designed for high-speed analysis of biological macromolecules, have been examined. Each of these columns is packed with a chemically modified silica gel, that has a controlled pore-size distribution and a uniform particle diameter. Surface silanol groups of these materials are chemically modified with hydrophilic polymers to produce a neutral silica surface, and ionic functional groups are chemically bonded to this neutral support, producing a stable ion exchanger. The separation of biological macromolecules on these columns has been investigated, and the selection of a mobile phase is discussed. PMID- 3031098 TI - Determination of adenosine ribo- and deoxyribonucleotides as their 1-N6-etheno derivatives by reversed-phase ion-pair high-performance liquid chromatography. AB - The chromatographic resolution of fluorescent 1-N6-etheno derivatives of adenine and adenosine ribo- and deoxyribonucleotides on reversed-phase columns has been optimised by a systematic study of the effect of ion-pair concentration, pH, eluent molarity and methanol concentration. Using tetrabutylammonium hydrogen sulphate as the ion-pair separation of up to eight derivatives could be achieved in ca. 10 min. Conditions of derivatisation with chloroacetaldehyde have been investigated in order to reduce hydrolysis of the nucleotides. PMID- 3031099 TI - Retention reproducibility of basic drugs in high-performance liquid chromatography on a silica column with a methanol-ammonium nitrate eluent. Interlaboratory collaborative study. AB - The reproducibility of the separation of basic drugs on silica columns has been tested in a collaborative study between nine laboratories. The results from the different laboratories were very similar. Various methods of recording the retention properties of the drugs were compared with reference to their reproducibility and ability to discriminate between different compounds. Relative retention times, relative capacity factors, and corrected capacity factors were much better than retention times and capacity factors, although all the methods suffered from large errors with weakly retained compounds. All the work was carried out using a single batch of silica to avoid variations due to the stationary phase. PMID- 3031100 TI - Comparison of bonded, polymeric and silica columns for chromatography of some penicillins. AB - The retention of ampicillin, amoxicillin, and penicillin G was determined on ODS and cyanopropyl (CN)-bonded to silica, on an all-organic polymeric reversed-phase column (styrene-divinylbenzene) and on bare silica. Solvents tested were 0.01 M H3PO4 (pH 1.6)-acetonitrile and 0.002 M KH2PO4 (pH 4.6)-acetonitrile in the proportions 100:0 to 10:90. Retention with 0.002 M KH2PO4 was weaker than with 0.01 M H3PO4 at all proportions of acetonitrile on all packings. With 0.01 M H3PO4, retention of penicillins on the polymeric column decreased rapidly from 100 to 50% aqueous phase and remained low to 10% aqueous phase. On the ODS bonded column, retention was similar, except that retention of the aminopenicillins, amoxicillin and ampicillin increased at less than 30% aqueous phase. On the CN bonded column, results were similar to those on the ODS column, except that the aminopenicillins were less strongly retained when using 100% 0.01 M H3PO4. On the bare silica, penicillin G was poorly retained at all proportions of acetonitrile, while retention of aminopenicillins increased rapidly at less than 50% 0.01 M H3PO4. Retention of aminopenicillins at less than 50% 0.01 M H3PO4 was parallel on bonded and bare silica but weaker on the bonded silica. A bonded organic layer weakened but did not prevent binding of amines to silica. PMID- 3031102 TI - Correction of retention index values in high-performance liquid chromatography as a tool for comparison of results obtained with different octadecyl silica phases. PMID- 3031101 TI - Rapid purification of tonin, esterase B, antigen psi and kallikrein from rat submandibular gland by fast protein liquid chromatography. AB - Tonin, esterase B, antigen psi and kallikrein from the rat submandibular gland were purified by fast protein liquid chromatography with Mono P or Mono Q columns. The purity of the separated proteins was evaluated by sodium dodecyl sulphate polyacrylamide gel electrophoresis and by isoelectrofocusing in flat-bed polyacrylamide gel. Tonin and esterase B were purified by DE-52 cellulose anion exchange chromatography and chromatofocusing on Mono P in two and three steps, respectively. Antigen psi and kallikrein were purified by a two-step procedure using DE-52 cellulose and Mono Q anion-exchange chromatography. The high resolution power of Mono Q revealed the different isoenzymes of kallikrein. PMID- 3031103 TI - Affinity fibre--a new support for rapid enzyme purification by high-performance liquid affinity chromatography. AB - A new type of support for chromatographic use has been developed. Nonporous quartz fibre, with a mean diameter of 0.5 micron, was silyated with mercaptopropyltrimethoxysilane. Tresyl chloride-activated dextran was covalently coupled to the SH groups on the fibre. Remaining active tresyl groups on the dextran were then coupled with an NAD derivative. The affinity fibre contained 0.3 mumol NAD derivative per g and was able to bind 15 mg of lactate dehydrogenase per g of fibre. The affinity fibre was used for large-scale purification of ox heart lactate dehydrogenase and its performance was compared with other commonly used chromatographic matrices. The operational capacity was found to be 1.0 g of pure lactate dehydrogenase per hour per 100 g opf fibre material (adsorption followed by salt elution). The affinity fibre was found to be particularly suitable for very rapid processing of large volumes of dilute enzyme solutions. PMID- 3031104 TI - High-performance liquid chromatographic determination of subsidiary dyes, intermediates and side reaction products in erythrosine. PMID- 3031105 TI - Improved method for the determination of sulphate in human serum using ion chromatography. PMID- 3031106 TI - Determination of itraconazole in plasma and animal tissues by high-performance liquid chromatography. PMID- 3031107 TI - A simple method for the visualization of the separated zones of sugars on silica gel TLC plates without spray reagent. AB - A simple procedure of visualization of the separated zones of sugars on silica gel TLC plates without spray reagent is described. The plate is placed into a glass chamber filled with n-hexane. The separated sugars are then seen as opaque zones. PMID- 3031108 TI - Antibody isotype response after human cytomegalovirus infection. AB - The antibody isotype response to human cytomegalovirus (CMV) was studied in paired sera from patients with primary and recurrent CMV infection. The majority of sera was obtained from immunocompromised patients. In the 48 patients with primary CMV infection, CMV-specific IgM (CMV-IgM) and IgG (CMV-IgG) antibodies were found in all patients, and CMV-specific IgA (CMV-IgA) and IgE (CMV-IgE) antibodies in 46 patients. CMV-IgM, -IgA, and -IgE antibodies were found in, respectively, 21, 31 and 4 of the 53 patients with recurrent CMV infection, and in, respectively, 3, 3 and 1 of the healthy controls. The results indicate that CMV-IgE is a better marker of primary CMV infection than CMV-IgM, and confirm that detection of CMV-IgM and -IgA may also be useful for diagnosis of recurrent CMV infection. PMID- 3031109 TI - Sensitivity and specificity of enzyme immunofiltration and DNA hybridization for the detection of HCMV-infected cells. AB - The sensitivity and specificity of enzyme immunofiltration and DNA hybridization were compared in human cytomegalovirus (HCMV) (AD 169)-infected MRC-5 cells. The enzyme immunofiltration was carried out on glass fiber filters in microplates, using an HCMV (AD 169) monoclonal antibody and a peroxidase conjugate. The DNA hybridization was carried out with a microfiltration apparatus, using a 32P labelled HCMV (AD 169) Eco R1 D fragment probe. The sensitivities of enzyme immunofiltration and DNA hybridization were 1.82 X 10(3) and 1.13 X 10(3) infected cells, respectively. Both methods were highly specific, but enzyme immunofiltration was faster and simpler. PMID- 3031110 TI - Detection of flavivirus RNA in infected cells using photobiotin-labelled hybridization probes. AB - Ten plasmids containing viral cDNA inserts of portions of the dengue virus type 2 (DEN-2) or Kunjin virus (KUN) genomes were biotinylated using photobiotin acetate and used as probes for the detection of flavivirus RNA in infected Vero cells. The viral cDNA inserts ranged in length from 0.19 to 2.7 kilobase pairs, and represented segments of the flavivirus genome coding for structural and nonstructural proteins. In spot hybridization assays (hybridization at 60 degrees C) with RNA extracted from cells infected with one of fourteen different flaviviruses or Semliki Forest virus, all DEN-2 and KUN probes hybridized specifically with RNA from cells infected with DEN-2 or KUN, respectively. At the reduced stringency of lower temperatures, specific hybridization to homologous viral RNA was still a feature of the probes, and only limited cross-hybridization to the RNA of some other flavivirus species was detected. PMID- 3031111 TI - Covalent binding of formalin fixed paraffin embedded brain tissue sections to glass slides suitable for in situ hybridization. AB - A novel method for covalently binding formalin fixed paraffin embedded (FFPE) tissue sections to glass microscope slides is validated suitable for in situ hybridization (ISH). Using the organosilane methodology of Maples (1985), 100% tissue adhesion is reported with no nonspecific probe binding, staining, or autoradiographic artefacts. JC viral nucleic acid sequences are successfully detected in FFPE progressive multifocal leukoencephalopathy brain tissue and the Tm of the hybridized product is estimated. From the Tm the most stringent washing condition resulting in an optimal signal to noise ratio is determined. A comparison is made between currently used methods of tissue adhesion and the proposed organosilane methodology. This methodology greatly facilitates studies of conditions for ISH and elucidation of mechanisms of viral infections requiring consecutive FFPE sections. It is also applicable to studies using cryosections and cultured cells. PMID- 3031112 TI - Competitive and blocking enzyme-linked immunoassay for detection of fetal bovine serum antibodies to bovine viral diarrhea virus. AB - A competitive blocking enzyme-linked immunoassay (CELIA) was developed to detect bovine viral diarrhea virus (BVDV) antibodies in undiluted fetal bovine serum (FBS). The CELIA was based on competition of serum BVDV antibodies with biotin labelled anti-BVDV immunoglobulins (Ig) for a limited quantity of solid-phase BVDV antigen. Antigen preparation was simple, FBS could be tested undiluted, and detergent-containing washes were unnecessary. A series of dilutions of postnatal bovine BVDV antiserum prepared in FBS and a set of 147 undiluted abbatoir FBS samples were tested by both CELIA and serum neutralization tests (SNT). CELIA results on both sets of specimens correlated positively with SNT titers (r = 0.99 and r = 0.85). Relative to the SNT, CELIA sensitivity was 100%; specificity was 76%. CELIA detected a level of BVDV antibody below the 1:2-titer threshold detectable with the SNT. Advantages, limitations, and theoretical differences between the CELIA and SNT are discussed. A similar comparison of CELIA with non competitive enzyme-linked immunoassay approaches to BVDV serodiagnosis is made. It is concluded that the CELIA is valuable in selecting only BVDV-seronegative FBS for use in virologic cell culture media. PMID- 3031113 TI - A protein immunoblot test for detection of bovine leukemia virus p24 antibody in cattle and experimentally infected sheep. AB - A protein immunoblot test for detecting antibody to the bovine leukemia virus p24 antigen is described. The test employs a crude antigen preparation derived from concentrated cell culture fluid, and an optimised biotin-streptavidin-peroxidase amplification system for immunodetection. The test is highly specific and is more sensitive than the gp51 agar gel immunodiffusion (AGID) test for detection of BLV antibody in cattle and experimentally infected sheep. In a selected set of 30 field sera from cattle which had given equivocal results in the gp51-AGID test, 21 were positive, 4 were negative, and 5 gave an uncertain result in the p24 immunoblot test. PMID- 3031114 TI - A test for human cytomegalovirus-specific immunoglobulins using a modification of a commercial test kit. AB - A technique (Ig-EIA) for the detection of CMV-specific IgG, IgM and IgA in human blood is described. Ig-EIA utilizes alkaline phosphatase-labeled goat anti-human IgG, IgM and IgA as a detection probe and CMV antigen-coated solid phase from commercial kits. Ig-EIA is compared to indirect fluorescent assay (IFA) and indirect hemagglutination (IHA) for sensitivity and specificity. On sequential samples of blood from a set of patients, Ig-EIA clearly demonstrated seroconversion in CMV-specific IgG and IgM. A test of 332 blood donors by Ig-EIA showed 177 (53%) had CMV-specific IgG and 17 (5%) had CMV IgM. Only two of the 17 donors with CMV IgM were nonreactive for CMV-IgG. The potential of CMV-IgM as an indicator of CMV infectivity is discussed. PMID- 3031115 TI - Cross-hybridisation of human papillomavirus DNA on filters. AB - Accurate type assignment of the different HPV types which infect the female genital tract, is essential in view of the differing pathological potential of the common virus types present in the cervix. We have developed hybridisation, washing and autoradiography conditions that minimise cross-hybridisation on filters and so allow clear-cut type assignment. We describe the conditions in this paper and have used this method to screen for HPV infection in clinical populations. PMID- 3031116 TI - Characterization of the twenty-four hour secretion patterns of adrenocorticotropin and cortisol in normal women and patients with Cushing's disease. AB - The episodic and circadian secretory patterns of ACTH and cortisol were studied in five patients with Cushing's disease (CD) and nine normal women by repetitive (every 20 min) venous blood sampling. In contrast to normal women, the 24-h transverse mean plasma ACTH levels were more than 3-fold greater (P less than 0.01), and the 24-h transverse mean serum cortisol levels were more than 2-fold higher (P less than 0.001) in the CD patients. Using a pulse detection algorithm, we found that the elevated ACTH levels in CD were accounted for, in part, by the more than 2-fold greater elevation in mean pulsatile ACTH amplitude (P less than 0.01), while the mean frequency of episodic ACTH and cortisol secretion was similar to that in normal subjects (10-12 episodes/24 h). Although a relatively close temporal relationship between ACTH and cortisol secretory episodes was found (r = 0.70 and 0.72, normal and CD groups, respectively), the increased ACTH pulse amplitude was not consistently associated with comparable cortisol pulses in CD patients. Circadian rhythms were identified for ACTH in two CD patients and for cortisol in all CD patients. The timing of the acrophases and nadirs for cortisol was not significantly altered compared to that in the normal group. After the noon meal, the normal postprandial elevation in cortisol was depressed or absent in the CD group. ACTH and cortisol responses to CRH in four CD patients were highly variable, but were not significantly different from those in normal subjects. These studies demonstrate that the elevated plasma ACTH levels in CD are sustained in part by increased ACTH pulse amplitude without significant alterations in pulse frequency. Despite the persistently high ACTH levels, the circadian variation of cortisol is maintained. The finding of an abnormal postprandial cortisol response in CD may provide an additional biological marker for CD. PMID- 3031117 TI - Receptors and actions of corticotropin-releasing hormone in the primate pituitary gland. AB - Receptors for CRH were identified in the pituitary gland of several primate species, and their binding characteristics were compared to the ability of CRH to elicit ACTH and cAMP responses in vitro. Autoradiographic analysis of the binding of [125I]Tyr-ovine CRH to frozen pituitary sections revealed CRH receptors in the intermediate and anterior lobes of human, marmoset, and cynomolgus monkey pituitaries. In the cynomolgus monkey, a high density of CRH receptors was present throughout the anterior and intermediate lobes. In the human pituitary, binding was concentrated in the anteromedial portion of the gland, whereas in the marmoset, binding was dense in the intermediate lobe and scattered as clusters throughout the anterior lobe. In membrane-rich fractions from the cynomolgus pituitary binding of [125I]Tyr-ovine CRH was time and temperature dependent, and was specific for CRH-related peptides; specific binding was increased by divalent cations and inhibited by guanyl nucleotides. Scatchard analyses of the binding data revealed a single class of high affinity sites [Kd, 1.93 +/- 0.23 (+/- SEM) nM], with a receptor concentration of 605 +/- 121 fmol/mg. In marmoset pituitary membranes, there were fewer receptors (200 +/- 15 fmol/mg), in agreement with the lower autoradiographic density of CRH binding. In anterior pituitary cell cultures from cynomolgus monkeys, CRH caused a dose-dependent stimulation of cAMP production and ACTH release, with half-maximum effective concentrations in the range of the CRH receptor affinity. Vasopressin and norepinephrine stimulated ACTH release to a much lesser extent, but both potentiated the stimulatory effect of CRH. Angiotensin II had no effect alone, but it also potentiated the effect of CRH. These data demonstrate the presence of CRH receptors in the primate pituitary, with characteristics similar to those in other species in their binding properties, coupling to adenylate cyclase, and functional interactions with other regulators of ACTH secretion that mediate the stimulatory effect of the peptide in the corticotroph. PMID- 3031118 TI - Tissues of the Laron dwarf are sensitive to insulin-like growth factor I but not to growth hormone. AB - Tissues from patients with Laron dwarfism are resistant to the actions of endogenous or exogenous GH. As a result, insulin-like growth factor I (IGF-I) levels are low, possibly contributing to the severe growth deficiency that occurs in patients with this syndrome. In this study, we found that erythroid progenitor cells and permanently transformed T-cell lines from two patients with Laron dwarfism responded in vitro to added IGF-I in concentrations ranging between 1-10 ng/mL despite no stimulatory response to added GH in concentrations of up to 500 ng/mL. Normal or near-normal responsiveness to insulin was also demonstrated. The persistence of GH resistance in the cultured T-cell lines confirms the primary genetic nature of the defect in Laron dwarfism. The preservation of in vitro growth responsiveness to IGF-I in hematopoietic tissue from the Laron dwarfs suggests that affected individuals are sensitive to this factor and may respond to it in vivo. PMID- 3031119 TI - Serum osteocalcin levels and bone alkaline phosphatase isoenzyme after oophorectomy and in primary hyperparathyroidism. AB - Serum osteocalcin levels peaked 1 yr after oophorectomy in a prospective study of 12 women. Estrogen treatment restored serum osteocalcin to the normal range within 4 months of therapy. The changes in serum osteocalcin preceded those in bone alkaline phosphatase activity by 1-2 months, in these oophorectomized patients and during estrogen treatment. The changes in these two markers of bone formation over time were significantly different from those in urinary hydroxyproline excretion. A significant positive correlation was found between bone alkaline phosphatase and serum osteocalcin levels in patients after oophorectomy and in 18 patients with primary hyperparathyroidism. Significant positive correlations also were found between the biochemical indices of osteoblastic function and urinary hydroxyproline excretion and/or nephrogenous cAMP in primary hyperparathyroidism. In most of the patients with primary hyperparathyroidism, however, the elevation in bone alkaline phosphatase was more marked than that in osteocalcin. These data indicate that the clinical utility of serum osteocalcin as a marker of bone formation is similar but not identical to that of bone alkaline phosphatase. PMID- 3031120 TI - A short negative feedback mechanism regulating corticotropin-releasing hormone release. AB - The effect of ACTH administration on plasma CRH levels was studied. In five patients with Addison's disease and three patients with hypopituitarism, bolus iv injection of 0.25 and 0.5 mg ACTH-(1-24) reduced plasma CRH levels (that had become elevated 48 h after discontinuation of corticosteroid replacement) to near normal levels at 30-60 min in a dose-dependent manner. Plasma immunoreactive beta endorphin levels were similarly decreased in patients with Addison's disease. ACTH-(1-24) (0.25 and 0.5 mg) injection failed to inhibit plasma CRH levels in five normal subjects. Basal CRH release from the rat hypothalamic median eminence in vitro was inhibited by 0.22 and 2.2 nM ACTH-(1-24) and ACTH-(1-39) in a dose dependent manner. These results suggest that in the absence of negative feedback control of ACTH secretion by glucocorticoids, ACTH can regulate its secretion by inhibition of hypothalamic CRH release. PMID- 3031122 TI - Transient hypoparathyroidism induced by synthetic human parathyroid hormone-(1 34) treatment. AB - Daily injections of low doses of a synthetic fragment of human PTH [hPTH-(1-34) have increased iliac trabecular bone volume when used in the treatment of osteoporosis. In approximately 50 patients no major side-effects had occurred. However, during daily sc 100-micrograms injections of the peptide, one patient repeatedly developed parathyroid hypofunction which resolved each time treatment was stopped. Specific immunoglobulin G (IgG) antibodies binding [125I]hPTH-(1-34) were identified in the patient's serum, and positive immunohistochemical reactions were obtained when bovine parathyroid sections were exposed to the patient's IgG. After adsorption with PTH, the patient's IgG, free of anti-PTH antibodies, reacted with renal cell membranes, as demonstrated by indirect immunofluorescence and blocked renal PTH-dependent adenylate-cyclase activation in vitro. These results support the hypothesis that anti-PTH receptor as well as anti-PTH antibodies were generated during hPTH-(1-34) treatment, which led to the development of hypoparathyroidism when their titers were high. PMID- 3031123 TI - Inhibitory effect of interferon-gamma on the response of human thyrocytes to thyrotropin (TSH) stimulation: relationship between the response to TSH and the expression of DR antigen. AB - Thyroid epithelial cells (thyrocytes) in autoimmune thyroid disease have been found to express DR antigens on their surfaces, and interferon-gamma (IFN gamma) induces DR antigen expression. This study was undertaken to determine the effect of IFN gamma on the response of human thyrocytes to TSH stimulation and the relationship between the response to TSH and the expression of DR antigen induced by IFN gamma, using monolayer cultures of Graves' thyrocytes. When confluent thyrocyte monolayers were incubated with TSH or Bu2cAMP (DBcAMP) for 7-9 days, T3 and thyroglobulin concentrations in the culture medium increased gradually in a dose-dependent manner. However, when TSH or DBcAMP was added after the cells had been cultured for 4 days with IFN gamma, T3 and thyroglobulin secretion in response to both 10 mU/mL TSH and 1 mM DBcAMP was significantly inhibited. The inhibition by IFN gamma was dose dependent and correlated with the number of DR antigen-positive thyrocytes present on the last day of culture. IFN alpha and beta did not affect the response of thyrocytes to TSH or DBcAMP stimulation. These results suggest that DR antigen-positive thyrocytes fail to respond to TSH stimulation at a site located distal to cAMP formation. PMID- 3031121 TI - Donor age-dependent decline in response of human red cell Ca2+-ATPase activity to thyroid hormone in vitro. AB - The effect of increasing donor age on the susceptibility of human red blood cell Ca2+-ATPase activity to stimulation in vitro by thyroid hormone was studied in 26 normal subjects, aged 15-81 yr. Basal enzyme activity (no added thyroid hormone) was unaffected by donor age. Group analysis, young (less than or equal to 50 yr) vs. elderly (greater than 60 yr old), revealed a 23% decrease in responsiveness of the enzyme to L-T4 (P less than 0.001). Regression analysis confirmed an age dependent decline in thyroid hormone stimulability of Ca2+-ATPase [r = -0.42 (T4 effect) and -0.38 (T3 effect); P less than 0.01]. Red cell membrane Na,K-ATPase activity was not affected by donor age. Plasma T4 and T3 concentrations in these normal subjects also did not change with age. Possible contributions of the following mechanisms to this age-correlated change in enzyme activity were examined: altered responsiveness to calmodulin of membrane Ca2+-ATPase; membrane content of endogenous calmodulin, endogenous plasma T4 and T3 concentrations, and plasma glucose concentrations. Calmodulin responsiveness is required for iodothyronine action on the enzyme, but the calmodulin responsiveness of cells from elderly donors was not significantly different from that of cells from younger donors (P greater than 0.10). There was no relationship between membrane immunoassayable calmodulin and donor age or membrane calmodulin and Ca2+-ATPase activity. There were positive correlations between donor plasma T4 level and basal enzyme activity (P less than 0.05) and between donor plasma T3 concentration and hormone-responsive Ca2+-ATPase (P less than 0.01), but these did not contribute to the age effect. Plasma glucose previously was found to modulate red cell Ca2+-ATPase activity, but did not correlate with decreased hormone responsiveness of the enzyme in elderly donors. In conclusion, we found that the susceptibility of human red cell Ca2+-ATPase to in vitro thyroid hormone stimulation declined significantly with advancing donor age. Several possible calmodulin-dependent mechanisms for this age-dependent change were excluded, and thus, we postulate that the altered hormone sensitivity of the enzyme is membrane phospholipid mediated. PMID- 3031124 TI - Responses of the hypothalamic-pituitary-adrenal and renin-angiotensin axes and the sympathetic system during controlled surgical and anesthetic stress. AB - We studied the responses of plasma CRH, ACTH, cortisol, norepinephrine, epinephrine, and renin activity in 11 patients undergoing parathyroid or thyroid surgery after identical preoperative sedation and during isoflurane (Forane) anesthesia. During surgical exploration, plasma CRH levels [10 +/- 2 (+/- SEM) pg/mL] remained at basal (unstimulated) levels, and plasma ACTH (11.5 +/- 1.4 pg/mL), cortisol (24 +/- 4 micrograms/dL), and epinephrine (35 +/- 10 pg/mL) concentrations remained within their normal morning ranges. The majority of the patients had no evidence of pulsatile ACTH secretion during the operation, but, rather, secreted ACTH and cortisol continuously. There was a small elevation of plasma norepinephrine and PRA which was associated with a small increase in heart rate and decrease in blood pressure. Anesthesia reversal, endotrachial extubation, and the early recovery period were associated with marked mean peak increases in plasma ACTH (173 +/- 45 pg/mL), cortisol (35 +/- 6 micrograms/dL), and epinephrine (220 +/- 56 pg/mL) and the return of plasma norepinephrine and PRA to basal levels. All hormones returned to basal levels by the first post operative day. The data suggest that with modern anesthetic techniques patients undergoing neck surgery had mildly elevated plasma ACTH, cortisol, and epinephrine levels. Glucocorticoid secretion during the operation was maintained primarily by continuous rather than pulsatile ACTH secretion. The immediate postoperative period was associated with profound elevations of plasma ACTH, cortisol, and epinephrine. The major determinant of ACTH, cortisol, and epinephrine secretion was anesthesia reversal and recovery and not surgical trauma. PMID- 3031125 TI - Pituitary-adrenocortical response to metoclopramide in patients with acromegaly and prolactinoma: a clinical evaluation of catecholamine-mediated adrenocorticotropin secretion. AB - We have demonstrated that metoclopramide stimulates cortisol secretion at least in part by a stress-mediated effect in normal men. To examine further the effect of the drug on the hypothalamo-pituitary adrenal system, we studied the cortisol response to 20 mg metoclopramide in patients with acromegaly, prolactinomas, and functional hyperprolactinemia and compared the results with the responses to insulin-induced hypoglycemia. In some patients, the effects of metoclopramide on CRH-induced ACTH and cortisol increase were studied to determine whether a change in dopaminergic (catecholaminergic) activity altered CRH stimulation of pituitary adrenal function. No cortisol response to 20 mg metoclopramide occurred in 13 tests on 8 of 9 patients with prolactinoma or acromegaly with hyperprolactinemia, whereas both acromegalic patients without hyperprolactinemia had a response. All of the patients had a normal cortisol response to insulin-induced hypoglycemia. Pretreatment with metoclopramide enhanced the CRH-induced cortisol increase from 30-120 min after CRH in normal men, but only at 15 and 30 min in 5 agromegalic patients. The results suggest that metoclopramide acts in the hypothalamus to release ACTH through a dopamine antagonist-mediated (catecholaminergic) mechanism, and that metoclopramide may act additively with CRH to stimulate ACTH secretion in normal men. The absence of a metoclopramide-induced cortisol response in patients with acromegaly or prolactinomas and the absence of a normal cortisol response to metoclopramide-CRH in acromegalic patients could be due to endogenous catecholamine deficiency in these patients. PMID- 3031126 TI - Antibodies against nuclear poly(A) polymerases in rheumatic autoimmune diseases. AB - Sera from 53 patients, 26 with systemic lupus erythematosus (SLE), 8 with rheumatoid arthritis (RA), 9 with Sjogren's syndrome (SS), and 10 with scleroderma (Scl), were screened for the presence of antibodies against liver type poly(A) polymerase and tumor-type poly(A) polymerase. Sixty percent of the patients with the above four autoimmune diseases have antibodies directed against liver poly(A) polymerase, whereas sera from 74% of the patients contained anti hepatoma poly(A) polymerase antibodies. About 25% of the patients produced antibodies exclusively against the tumor poly(A) polymerase. IgG containing anti liver or anti-tumor poly(A) polymerase antibodies inhibited the activity of the respective enzyme. IgG containing antibodies against liver and tumor enzymes inhibited the activity of both enzymes, whereas IgG from sera that did not react with poly(A) polymerase had no effect on either enzyme. These data demonstrated the specificity of these autoantibodies and confirmed the results of the radioimmunoassay. PMID- 3031127 TI - Antiprogesterone activity of RU 486 and its contragestive and other applications. AB - RU 486 is the first antiprogesterone to be used clinically. It inhibits the action of the hormone at the receptor level in target tissues. Its action is particularly significant in the endometrium where it prevents the initiation and progression of pregnancy in the first weeks (contragestive effect). The data indicate that the compound can be used for: voluntary interruption of pregnancy between 6 and 10 weeks; induction of menstruation during the fifth week of amenorrhoea, and post-coital contraception. Current trials include its use as a once-a-month menses inducer. It can also be utilized for therapeutic interruption at a late stage of pregnancy, and tried as adjuvant treatment in some cases of breast cancer. The data on RU 486 have been obtained through studies in physio pharmacological endocrinology and biochemistry. The development of this antihormone represents a concerted research link between biology and medicine. PMID- 3031128 TI - Human luteolysis--interaction between HCG and oestradiol-17beta in an in-vitro model. AB - Corpora lutea (CL) were excised from each of 24 women undergoing laparotomy at different and well-classified stages of the luteal phase of the menstrual cycle. Pieces were incubated for intervals between 30 min and 4 h, the incubations being carried out in the presence or absence of human chorionic gonadotrophin (HCG) and oestradiol-17beta (E2) along and in combination. Following incubation, the tissue levels of cAMP and the media concentrations of progesterone and, in certain cases, pregnenolone, were determined. HCG stimulated the production of cAMP and progesterone in CL of all ages. This effect on the formation of progesterone was significantly counteracted by E2 in CL of the early and mid-luteal phases. The addition of indomethacin to the incubation media did not alter this anti gonadotrophic effect of E2 in-vitro. The formation of cAMP was not influenced by E2 in-vitro. These results suggest that the well-known luteolytic effect of E2 in vivo is caused by inhibition of the gonadotrophin-stimulated synthesis of progesterone by corpora lutea, an effect mediated neither by prostaglandins nor cAMP. PMID- 3031129 TI - Changes in cerebrospinal fluid cells, IgG and IgA during herpes simplex virus encephalitis in rabbits. AB - Rabbits were infected with herpes simplex-type 1 virus either by corneal scarification or intrathecal inoculation. Encephalitis was induced predictably by either route but was most severe after intrathecal inoculation. Serial examination of the cerebrospinal fluid (CSF) demonstrated abnormalities reflecting two distinct phases of the immune response to this central nervous system infection. The acute phase was manifested by a mononuclear pleocytosis and transudation of serum proteins into the CSF. The recovery phase was manifested by increased amounts of IgG, IgA and antibody specific for herpes simplex virus in the CSF. These studies demonstrate that IgA is a significant component of the local immune response to viral encephalitis in the rabbit as well as in mice and man. PMID- 3031130 TI - Analysis of the intrathecal humoral immune response in Brown Norway (BN) rats, infected with the murine coronavirus JHM. AB - Serum and CSF specimens from clinically healthy Brown Norway (BN) rats inoculated intracerebrally with corona virus JHM were analysed with respect to the state of the blood-brain barrier (BBB) and the intrathecal synthesis and isoelectric distribution of immunoglobulins (Ig). Increased CSF/serum ratios for Ig in the context of an intact BBB were never seen in the absence of intrathecal synthesis of virus-specific antibodies. Affinity-mediated immunoblot analysis revealed a broad pattern of virus-specific antibodies with embedded clusters of restricted heterogeneity, but no signs of oligoclonal Ig production carrying non-viral specificity. From these data it was concluded that BN rats do control the intracerebral spread of JHM virus effectively by a strong local virus-specific antibody response, thereby preventing a clinically apparent disease. PMID- 3031131 TI - Uptake of oleate from albumin solutions by rat liver. Failure to detect catalysis of the dissociation of oleate from albumin by an albumin receptor. AB - The hepatic removal of albumin-bound substances from plasma requires that they dissociate from albumin. Using indirect methods, we and others have proposed that dissociation may be catalyzed by interaction of albumin with the liver cell surface. This study looked for direct evidence of catalysis by comparing the rate of dissociation of oleate from albumin in vitro with the rate observed within the sinusoids of perfused rat liver. No evidence for catalysis was found. The rate of hepatic oleate removal from dilute albumin solutions did not exceed but instead closely paralleled the rate predicted from the in vitro dissociation rate constant (0.14s-1). These results suggest that under some conditions the liver can remove unbound material from the sinusoids faster than it can be replenished by dissociation from albumin, resulting in dissociation-limited removal. However, dissociation of oleate does not appear to be catalyzed by the liver. PMID- 3031132 TI - Monoclonal antibody characterization of a chymotrypsin-like molecule on neutrophil membrane associated with cellular activation. AB - Monoclonal antibody 1-15 (Ab 1-15), is a murine anti-human neutrophil (PMN) IgG1 that inhibits PMN effector responses to N-formyl-met-leu-phe (FMLP) and phorbol myristate acetate. In this study, the effects of Ab 1-15 on PMN membrane-related functions were characterized: Ab 1-15 inhibited PMN superoxide (O-2) response to FMLP by 60% (P less than 0.005) without effect on the onset or duration of O-2 production. This inhibition of O-2 response was associated with a significant inhibition of PMN chymotrypsin-like, but not trypsin-like, protease activity. Cell fractionation studies indicated the presence of an Ab 1-15 inhibitable, chymotryptic neutral protease activity in PMN membranes. In studies of Ab 1-15 effects on membrane-related second messenger pathways, Ab 1-15 augmented both FMLP- and isoproterenol-induced intracellular cAMP accumulation, whereas alpha chymotrypsin decreased PMN cAMP response to these stimuli. Our data suggest that the function-inhibiting, anti-PMN monoclonal Ab 1-15 defines a PMN chymotryptic enzyme on the membrane surface that is involved in regulation of two membrane related functions, O-2 generation and cAMP generation. PMID- 3031133 TI - Growth advantage and enhanced toxicity of Escherichia coli adherent to tissue culture cells due to restricted diffusion of products secreted by the cells. AB - This study was undertaken to examine whether Escherichia coli adherent to tissue cells gain advantages over nonadherent bacteria due to their proximity to the cells. We used tissue culture cells and isogenic derivatives of a proline auxotrophic strain of E. coli that were fimbriated (Fim+) or nonfimbriated (Fim ), and were heat-labile enterotoxin producing (Tox+) or toxin nonproducing (Tox ). We found that the Fim+ bacteria; which were capable of adhering to tissue culture cells, initiated growth much sooner than did nonadherent Fim- bacteria; the adherent bacteria used tissue cell-derived proline, which was available at high concentrations only in the zone of bacterial adherence. Likewise, cyclic AMP secreted by adherent (Fim+) bacteria was maintained at high concentration on the tissue cell surfaces. As few as 2 X 10(5) adherent Fim+ Tox+ bacteria exert toxic activity upon Y1 adrenal cells, whereas toxin secreted in the medium by 6 X 10(6) Fim- Tox+ bacteria was undetectable. The results suggest that the growth advantage and enhanced toxicity of adherent E. coli is due to restricted diffusion of products secreted by the tissue culture and bacterial cells, respectively. PMID- 3031134 TI - Influence of Cl- on organic anion transport in short-term cultured rat hepatocytes and isolated perfused rat liver. AB - Transport of 35S-labeled sulfobromophthalein [35S]BSP was studied in short-term cultured rat hepatocytes incubated in bovine serum albumin. At 37 degrees C, initial uptake of [35S]BSP was 5-10-fold that at 4 degrees C, linear for at least 15 min, saturable, inhibited by bilirubin, and reduced by greater than 70% after ATP depletion or isosmotic substitution of sucrose for NaCl in medium. Replacement of Na+ by K+ or Li+ did not alter uptake, whereas replacement of Cl- by HCO-3 or gluconate- reduced uptake by approximately 40%. Substitution of Cl- by the more permeant NO-3 enhanced initial BSP uptake by 30%. Efflux of [35S]BSP from cells to media was inhibited by 40% after ATP depletion or sucrose substitution. To confirm these results in a more physiologic system, transport of [3H]bilirubin was studied in isolated livers perfused with control medium or medium in which Cl- was replaced by gluconate-. Perfusion data analyzed by the model of Goresky, revealed 40-50% reductions in influx and efflux with gluconate- substitution. These results are consistent with existence of a Cl-/organic anion exchange mechanism similar to that described by others in renal tubules. PMID- 3031136 TI - Prediction of human papilloma virus antigen in cervical squamous epithelium by koilocyte nuclear morphology and "wart scores": confirmation by immunoperoxidase. AB - Koilocytes (balloon cells) in cervical squamous epithelium can be distinguished by their nuclear morphology as members of two populations A and B. The proposition that population A was infected with human papilloma virus (HPV) and population B was not, was examined immunohistologically. A peroxidase antiperoxidase technique using polyclonal HPV antibody failed to support the hypothesis and showed small fractions of both populations to be infected with the virus (A = 5 of 25; B = 2 of 19). Nuclear morphology alone is thus inadequate to distinguish infected from non-infected koilocytes, or balloon cells. When a number of well established histological changes in squamous epithelia infected with HPV were examined, graded, and summated to obtain a "wart score," however, a reasonably accurate prediction of HPV infection emerged. PMID- 3031137 TI - Nerve growth factor (NGF) receptor expression in chicken cranial development. AB - In order to map the expression of receptors for nerve growth factor (NGF) during brain and cranial ganglia development, iodinated NGF (125I beta NGF) was used as a probe in an autoradiographical analysis performed between embryonic day 3 (E3) and posthatching day 3 (P3) of chicken development. Heavy autoradiographic labelling was observed at the classical NGF target sites, the proximal cranial sensory ganglia and the sympathetic superior cervical ganglion, throughout development and after hatching. In contrast, only weak labelling could be detected during a restricted time span in the vestibulocochlear (E4-E8) and the distal cranial sensory ganglia (E4-E10), the neurons of which originate from the otic and epibranchial placodes. Specific 125I beta NGF binding was also observed in various brain regions during early brain development. NGF receptor expression there followed a characteristic pattern. The neuroepithelial layer displayed very low levels of specific 125I beta NGF binding, while strong 125I beta NGF labelling was found in the mantle layer. Brainstem somatomotor nuclei, visceromotor columns, brainstem alar plate, cerebellar anlage, tectum, and basal forebrain (epithalamus, striatum) were found to be transiently labelled by 125I beta NGF in early development (E4-E12). Non-nervous tissues such as parts of the otic vesicle epithelium and skeletal muscle anlagen of the head were also labelled. These results, showing specific binding of 125I beta NGF to cranial cells of different origin (neural tube, neural crest, placode, and possibly mesoderm) strengthen the concept that NGF may have diverse functions in growth and differentiation of various tissues and cell types. PMID- 3031135 TI - Inositol 1,4,5-triphosphate-induced granule secretion in platelets. Evidence that the activation of phospholipase C mediated by platelet thromboxane receptors involves a guanine nucleotide binding protein-dependent mechanism distinct from that of thrombin. AB - Phosphoinositide hydrolysis in platelets stimulated by thrombin is thought to be regulated by a pertussis toxin-sensitive guanine nucleotide binding protein (G protein) referred to as Gp. The present studies examine the role of Gp in platelet responses to the thromboxane A2 analogue U46619 and in the pathway by which the phosphoinositide hydrolysis product inositol 1,4,5-triphosphate (IP3) causes secretion. In permeabilized platelets, U46619 caused phosphatidic acid formation and secretion, which were abolished by the G protein inhibitor, guanosine 5'-O-(2-thiophosphate) (GDP beta S). Unlike thrombin, however, U46619 induced phosphoinositide hydrolysis was unaffected by pertussis toxin, and U46619 was unable to inhibit the [32P]ADP ribosylation of the 42-kD pertussis toxin substrate in platelets. IP3-induced secretion, which is known to depend upon intracellular Ca release and subsequent arachidonic acid metabolism, was also inhibited by GDP beta S, as was Ca-induced secretion. These observations suggest that platelet thromboxane A2 (TxA2) receptors are coupled to a toxin-resistant form of Gp distinct from the one that is coupled to thrombin receptors, and that TxA2-stimulated phosphoinositide hydrolysis may serve as a feedback mechanism by which stimuli for arachidonic acid release, such as IP3 and Ca, amplify responses to agonists. PMID- 3031138 TI - A malignant nephroblastoma in an aged fox (Fennecus zerda). AB - A malignant nephroblastoma with pulmonary metastasis which was found at necropsy in an old fox is described. This is the first report of such a tumour in a fox. Nephroblastoma is rare in the family Canidae and usually occurs in young individuals. The presence of a tumour arising from embryonal tissue in an aged animal raises questions about the genesis and behaviour of this tumour. The tumour may have contributed to the animal's congestive heart failure as a result of the generalized pulmonary involvement, an erythropoietin-induced polycythaemia, or increased peripheral resistance via the renin-angiotensin system. PMID- 3031139 TI - Relationship between cellular morphology and immunocytological findings of spontaneous pituitary tumours in the aged rat. AB - A total of 339 spontaneous pituitary tumours from 284 aged rats were studied by light microscopy. Based on the cellular morphology in preparations stained by haematoxylin and eosin, tumour cells were classified into 4 types: granular; agranular small; agranular large polygonal; and agranular large slender cells. Eight tumours consisted of mixed neoplastic masses of 2 types of cells. A total of 85 cases representative for each tumour type was used for special staining and immunocytochemistry. All tumours of the granular cell type and some tumours of the agranular large polygonal cell type had granules which stained red by azocarmine and were positive for prolactin. All agranular small cell type tumours were also positive for prolactin and agranular slender cell type tumours were positive for adrenocorticotropic hormone. Only tumours of the agranular large polygonal cell type gave an indefinite or negative reaction for all the various pituitary hormones. These results indicate a relatively good correlation between the cellular morphology and the immunocytochemistry of spontaneous pituitary tumours in aged rats. PMID- 3031141 TI - Relationship between milkability and adrenoceptor concentrations in teat tissue in primiparous cows. AB - Milking characteristics were measured on 19 primiparous cows of the Red Pied breed at morning milkings. Measurements included maximum and average rate of flow, yield of milk through the 1st, 2nd, and 3rd min, amount of milk through the first 2 and first 3 min of milking, and milking time. Subsequently cows were slaughtered and teats immediately removed. Adrenoceptors on membranes isolated from teat tissue were identified by radiolabeled antagonists: [3H]dihydroalprenolol for beta 2-adrenoceptors, [3H]rauwolscine for alpha 2 adrenoceptors, and [3H]prazosin for alpha 1-adrenoceptors. Measurements of milking characteristics were highly repeatable within cow. Cows showed five distinct and significantly different milk flow patterns, which were characteristic for fast (type I, II), relatively fast (type III), and slow (type IV, V) milking cows. Covariance analysis for the five different types revealed an average regression between alpha 2-adrenoceptor density and milking rate 1st min as well as between beta 2:alpha 2-adrenoceptor ratios and milking rate 2nd min and milking time. A hypothesis is presented to explain these observations. During milking, tone of efferent sympathetic nerves in the teat is low. This phenomenon is more pronounced in fast milking cows. The typical presynaptic adrenoceptor pattern in the teat of fast milkers (low beta 2:alpha 2-adrenoceptor ratio) results in a decline of norepinephrine release by feedback mechanisms. PMID- 3031140 TI - Malignant transformation of eccrine tumors. AB - Malignant transformation occurred in pre-existing sweat gland tumors in 7 patients. Three lesions showed an histologic pattern of eccrine spiradenoma, 2 eccrine poroma, one cylindroma and one papillary eccrine adenoma. Malignant transformation was histologically characterized by the presence of solid tumor areas populated with large cells having irregularly shaped nuclei and mitotic figures. There were multiple foci of squamous metaplasia, areas of loss of basement membrane and invasion of the surrounding connective tissue. PMID- 3031143 TI - A case of poroma folliculare on the forearm. PMID- 3031142 TI - Syringocystadenoma papilliferum associated with poroma folliculare. PMID- 3031144 TI - Platelet [3H]imipramine binding in depressed patients and its circadian variations in healthy controls. AB - Platelet [3H]imipramine binding was determined in 28 patients with major depression, 11 with bipolar disorders, and 28 healthy controls. The mean maximum number of binding sites (Bmax) in depressed patients was significantly lower than in healthy controls. A significant negative correlation was found between the Bmax values and the total scores of the 17-item Hamilton depression rating scale in major depression. Our results suggest that the Bmax values in major depression may be related to severity of depression. There were significant circadian variations in the Bmax values of [3H]imipramine binding on human platelets from six healthy controls. The mean Bmax values were significantly low in the dark phase and high in the light phase. PMID- 3031146 TI - Histologic and biochemical correlates of left ventricular chamber dynamics in man. AB - To investigate the relation between left ventricular chamber dynamics in humans and the quantitative analysis of the histologic and biochemical characteristics of left ventricular endomyocardial biopsy material, 15 patients with a wide range of ventricular function were studied. The pressure-volume relation was determined using simultaneous gated radionuclide angiography, echocardiography and micromanometer pressure. The derived chamber dynamics were then compared with quantitative histologic data (percent fibrosis and cell diameter) and adenosine triphosphate content measurements obtained from the left ventricular biopsy specimen obtained at the time of the pressure-volume studies. The measures of systolic function correlated linearly with high energy phosphate content. The adenosine triphosphate/protein ratio (nanomoles) was shown to parallel ejection fraction (r = 0.81), peak ejection rate (r = -0.73) and peak positive maximal rate of rise in left ventricular pressure (dP/dt) (r = 0.79). No correlation was observed between these variables and the percent fibrosis or cell diameter. Variable results were found in comparing the diastolic properties of the left ventricle with the biopsy data. In general, the high energy phosphate content correlated with measures of active relaxation, but not with the passive filling characteristics of the left ventricle. The adenosine triphosphate/protein ratio was linearly related to peak negative dP/dt (r = -0.74) and the peak filling rate (r = 0.76) but correlated less well with other measures of active and passive diastolic filling. No correlation was found between any diastolic variable and the percent fibrosis or cell diameter.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3031145 TI - Adrenocortical responsiveness to the ACTH stimulation test in depressed patients and healthy volunteers. AB - Adrenocortical activation in depression has been postulated to result from overactivity of limbic system-hypothalamic function. However, some studies suggest the possibility that excessive secretion of cortisol might result, in part, from a heightened adrenocortical responsiveness to ACTH. To further examine this possibility, we utilized both the ACTH stimulation test and the overnight dexamethasone suppression test (DST) in 72 patients with major depression and 37 age- and gender-matched healthy volunteers. The melancholic/DST-nonsuppressor group had larger mean peak cortisol and cumulative cortisol responses (CCR) than any of the other patients groups or healthy controls. However, the differences failed to reach statistical significance as a result of a relatively large cortisol response variability. Nevertheless, the present findings are in general agreement with previous reports suggesting the possibility of an enhanced adrenocortical responsiveness to ACTH. PMID- 3031147 TI - Lipid-derived and other chemical mediators of inflammation in the lung. PMID- 3031148 TI - Isolation of the C1r sub-unit of the first complement component (C1), avoiding low pH values. PMID- 3031149 TI - Effects of prenatal maternal stress on the pituitary adrenocortical reactivity in guinea-pig pups. AB - Seven days before term, pregnant guinea-pigs were subjected to a psychosomatic stress. Adrenocorticotrophin (ACTH), cortisol and aldosterone concentrations increased in maternal and fetal plasma immediately after stress. In the offspring born from prenatally-stressed mothers that were resubjected to stress, the increases in plasma ACTH and cortisol level were lower than in the control offspring. Plasma aldosterone levels increased after stress in 12 days-old pups but did not change neither in 50 days-old animals nor in 12 days-old guinea-pigs born of prenatally stressed mothers. Thus prenatal stress of mother resulted in lowered response of pituitary-adrenocortical axis of offspring subjected to stress. PMID- 3031150 TI - Predicting the outcome of corticoid therapy for acute ulcerative colitis. Results of a prospective, randomized, double-blind clinical trial. AB - We looked for factors predicting the therapeutic outcome in 66 patients with severe ulcerative colitis treated with intravenous hydrocortisone or corticotropin (ACTH) for 10 days. Patients were randomized before therapy within strata defined by whether they had received oral corticosteroids continuously before the study (group A, 35 patients) or not (group B, 31 patients). Comparisons were made between groups receiving what we considered optimal corticoid therapy, hydrocortisone for group A and ACTH for group B. Overall, therapeutic success was achieved in 28 (42%), with a median time of 7.5 days. Favorable factors measured on admission to the study were those suggesting less severe colitis activity: absence of fulminant disease, limited disease extent, a shorter duration of the present attack, fewer stools, a lower erythrocyte sedimentation rate (ESR), and a higher hemoglobin. Factors compatible with more severe colitis including fulminant activity, more extensive disease, a shorter total disease duration, bloody stools, and fewer bowel movements, favored an early response among those patients who were to achieve a remission. Prolonging therapy beyond 10 days by switching to the alternate corticoid drug did not improve the remission rate. Achieving remission during the initial therapy period, especially when it occurred early, was the most important predictive factor for a favorable clinical course during the following year. Prolonging therapy did not improve the 1-year remission rate. In fact, a higher proportion of patients who continued to require therapy underwent colectomy than those who received one treatment course. PMID- 3031151 TI - Preponderance of serum and intra-hepatic 5 alpha-dihydrotestosterone in males with hepatocellular carcinoma despite low circulating androgen levels. AB - To investigate possible influences of the sex-steroid milieu on hepatocellular carcinoma (HCC) and vice versa, circulating and intra-hepatic sex-steroid levels were investigated and compared with the levels in cirrhosis alone. In cirrhotic men with HCC, serum 17 beta-oestradiol levels were normal, unlike the elevated levels in men with cirrhosis alone. Total and free levels of testosterone and 5 alpha-dihydrotestosterone (DHT) were lower in patients with HCC than in cirrhotic or normal men; the greater decrease in testosterone levels caused an elevated DHT: testosterone ratio. Hypothalamic-pituitary axis dysfunction demonstrated for HCC and cirrhotic groups could not explain the differences in sex-steroids between them. Compared with normal tissue, HCC cytosol had lower testosterone and similar DHT levels; both androgens were higher than in cirrhotic tissue, and the intracellular DHT: testosterone ratio in the tumour was much higher than in control tissue. Results suggest alterations in sex-steroid metabolism in HCC favouring hepatic accumulation of 5 alpha-reduced metabolites aided by the elevated intracellular sex-hormone binding globulin levels shown in HCC tissue. PMID- 3031152 TI - Antibody to the hepatitis B virus receptor for polymerized albumin in acute infection and in hepatitis B vaccine recipients. AB - A receptor for polymerized human serum albumin is encoded by the pre-S region of the hepatitis B virus genome and may mediate attachment of the virion to hepatocytes. To investigate antibody response to the virus receptor we studied sera and their IgG fractions for inhibitory activity on hemagglutination of polyalbumin-coated red cells by virus particles containing the pre-S polypeptide. By this method antibody to the receptor was detected in serum in a goat immunized with pre-S containing particles, with no relation to levels of antibody to hepatitis B surface antigen, and in the sera of 33% and 83%, respectively, of acute hepatitis B patients studied during the early phase of illness and during convalescence. In contrast, antibody to the receptor was not detected in serum in any of the 47 subjects immunized with a commercial, plasma-derived, hepatitis B vaccine. These results demonstrate that natural acute infection with hepatitis B virus leads to production of antibody to the virus receptor for polyalbumin, while such antibody response is absent after immunization with currently licensed hepatitis B vaccines. PMID- 3031153 TI - Further progress towards understanding hepatic sinusoidal cells. PMID- 3031154 TI - Noradrenergic mechanisms in the response to dextroamphetamine in humans. AB - The effect of oral thymoxamine, an alpha-noradrenergic antagonist, on the arousal and anorectic action of oral dextroamphetamine was measured in order to investigate the role of the alpha-noradrenergic pathways in these responses. Thymoxamine appeared to accentuate the alerting action and reduce the anorectic action of dextroamphetamine. The implications of these findings on the role of the alpha-noradrenergic pathways in these responses is discussed in the light of previous findings. PMID- 3031155 TI - Distribution of basal lysosomes in exocrine acinar cells. AB - We examined the distribution of trimetaphosphatase (TMPase)-positive basal lysosomes in pancreas, parotid, submandibular, sublingual, and exorbital lacrimal glands from rats, rabbits, and guinea pigs. The location of the basal lysosomes was compared to that of the acid phosphatase (AcPase)-positive lysosomes. In all of the tissues examined from rat and rabbit, AcPase activity was localized primarily to the Golgi region. Reaction product was localized in GERL, immature secretory granules, and lysosomes lying adjacent to the Golgi apparatus. TMPase activity was found in basal lysosomes and in occasional elongated lysosomes adjacent to the Golgi apparatus. In guinea pig, the distribution of TMPase activity was identical to that seen in the other two species, but a significant number of lysosomes in the basal region of the cells also contained AcPase activity. These results confirm and extend our previous finding (J Histochem Cytochem 31:1209, 1983) that exocrine acinar cells possess two distinct populations of lysosomes. The lysosomes in the Golgi region contain both AcPase and TMPase activity, whereas those in the basal portion of the cells are reactive predominantly for TMPase. The functional significance of the two populations of lysosomes is not understood at present. PMID- 3031156 TI - Cerium-based cytochemical method for detection of ouabain-sensitive, potassium dependent p-nitrophenylphosphatase activity at physiological pH. AB - We have developed a new cytochemical method for detecting the ouabain-sensitive, potassium-dependent p-nitrophenylphosphatase (K-NPPase) activity of the sodium potassium-activated adenosine triphosphatase (Na-K ATPase) complex. The incubation medium contains p-nitrophenylphosphate (p-NPP) as substrate, cerium chloride as capture agent, Tricine buffer, MgCl2, and KCl. Tricine buffer protected against the medium turbidity caused by non-enzymatic reaction at pH 7.5. Biochemically, the accumulation of p-nitrophenol and phosphate in the reaction precipitate was proportionally related to the enzyme concentration. Ultracytochemically, the reaction products of the K-NPPase activity were localized as fine and uniform electron-dense deposits in the cytoplasmic side of specialized basolateral plasma membranes of cells of kidney distal convoluted tubules, secretory cells of salt gland, and marginal cells of stria vascularis. This method has the advantage of being useful at physiological pH. PMID- 3031157 TI - Superinduction of T lymphocyte-endothelial cell (EC) binding by treatment of EC with interleukin 1 and protein synthesis inhibitors. AB - We have previously reported that marked enhancement of the in vitro binding of lymphocytes to endothelial cell (EC) monolayers is observed after stimulation of the EC with interleukin 1 (IL 1). To determine whether new protein synthesis was required for this effect of IL 1, EC were incubated with IL 1 in the presence of cycloheximide or puromycin. Three different effects of these protein synthesis inhibitors on T-EC binding were observed. First, preincubation of the EC with both IL 1 and an inhibitor blocked the increase in binding if the inhibitor was present during both the preincubation and the 1 hr duration of the T-EC binding assay, suggesting that new protein synthesis is required for the enhancement of T EC adhesion by IL 1. Second, preincubation of the EC with low doses of the inhibitors (0.1 to 1 microgram/ml) in the absence of IL 1 consistently increased T-EC binding, even if the inhibitors were present during the T-EC adhesion assay; in addition, the inhibitors additionally increased the stimulatory effect of IL 1 if the EC were washed free of the inhibitor before the assay step. The binding enhancing effect of low concentrations of cycloheximide could be inhibited by an antibody to the CDw18 complex on the T cell, suggesting an up-regulation of the ligand on the EC involved in CDw18-dependent T cell adhesion. Third, higher concentrations of the inhibitors (3 to 10 micrograms/ml) were toxic for the EC in the presence of IL 1, possibly due to the combined blocking effect of IL 1 and inhibitors on EC protein synthesis. PMID- 3031159 TI - Autoantibodies specific for the cardiac myosin isoform are found in mice susceptible to Coxsackievirus B3-induced myocarditis. AB - Several mouse strains are susceptible to immunopathic myocarditis after infection with Coxsackievirus B3 (CB3). This disease is associated with autoantibodies that are directed against myosin. In this study we characterized sera from CB3 infected mice for their reactivity with three different myosin isoforms (heart, skeletal muscle, and brain myosins) and for autoantibody isotype by using an ELISA. Competitive inhibition assays and absorption studies with various myosins demonstrated the presence of two autoantibody populations in sera of susceptible A.CA and A.SW mice. The first was specific for cardiac myosin and was mainly IgG. The second antibody population cross-reacted with heart, skeletal muscle, and brain myosin and was mainly IgM. B10.PL/SgSf and B10.A/SgSf mice, which do not develop immunopathic myocarditis, produced only the IgM autoantibody population cross-reactive with all three myosin isoforms. Because the heart-specific myosin autoantibodies were found exclusively in the mouse strains that developed immunopathic myocarditis, they can be considered a serologic marker for autoimmune heart disease. PMID- 3031158 TI - Oxidation of salicylates by stimulated granulocytes: evidence that these drugs act as free radical scavengers in biological systems. AB - Although salicylates have been used for centuries as treatment of inflammatory diseases, the mechanism of action of these drugs is still not clear. Aspirin (acetylsalicylic acid) and other nonsteroidal anti-inflammatory drugs (NSAID) inhibit prostaglandin biosynthesis, a property that appears to explain part of their anti-inflammatory activity. However, this mechanism does not appear to explain the anti-inflammatory properties of salicylic acid, which is a major metabolite of ASA in vivo. Results of prior studies in our laboratory have established that benzoic acid, the parent compound of the salicylate group of drugs, is decarboxylated and hydroxylated by the hydroxyl free radical (OH.) produced by stimulated granulocytes. These observations suggested that salicylates might be similarly metabolized by granulocytes. If so, the capacity of salicylates to rapidly react with OH. might relate directly to their known anti-inflammatory properties. Preliminary experiments established that salicylic acid and aspirin were decarboxylated by the hydroxyl free radical generated by the enzyme system xanthine-xanthine oxidase. We then studied the metabolism of salicylates by human granulocytes. Unstimulated granulocyte suspensions did not oxidize ASA or salicylic acid. However, suspensions stimulated by opsonized zymosan particles rapidly oxidized both substrates in pharmacological concentrations. The rate of oxidation of salicylic acid was 16-fold higher than benzoic acid, whereas the rate of oxidation of ASA was four-fold higher. The reaction was oxygen dependent and could be inhibited by known hydroxyl scavengers, particularly dimethylthiourea. The reaction could also be inhibited by superoxide dismutase and azide, indicating that O-2 and heme or an iron dependent enzyme were required for the reaction. The reaction was not impaired by compounds known to react with the HOCL and the chloramines generated by stimulated PMN. Furthermore, salicylic acid in high concentrations did not impair the HMPS pathway, the production of O-2 or the production of H2O2 by granulocytes. These data provide evidence that salicylates are rapidly oxidized by the hydroxyl free radical produced by granulocytes and not O-2, H2O2, or HOCL. This capacity of salicylates to react rapidly and selectively react with OH. may directly relate to their anti-inflammatory properties. In addition, results of our experiments indicate that stimulated granulocytes acquire the capacity to metabolize these drugs. Therefore, several metabolites of salicylates may be produced at a site of inflammation, all of which may have altered biological activity compared with the parent compound. PMID- 3031160 TI - Development and characterization of a human B cell line that responds to B cell growth factor but not interleukin 2. AB - The human lymphoblastoid cell line we present here proliferated in response to a 14,000 m.w. B cell growth factor (BCGF), and not to interleukin 2 (IL 2). This cell line, designated B-A3, was established by Epstein Barr virus (EBV) transformation of Staphylococcus aureus Cowan I (SAC)-activated spleen B cells, and has been maintained in RPMI 1640 medium complemented with 15% fetal calf serum (FCS) without the addition of other exogenous growth factors. A proliferative response, as measured by [3H]thymidine uptake of B-A3 cells was significantly induced by either commercial IL 2-free human BCGF preparations, or phytohemagglutinin-stimulated mixed lymphocyte culture supernatant at all FCS concentrations used in the assay. The most marked proliferation due to BCGF, however, was observed in the absence of FCS. This BCGF-induced proliferation was not influenced by IL 2 or interferon-gamma (IFN-gamma), because both recombinant IL 2 and IFN-gamma failed to induce proliferation. The response of B-A3 cells to a specific BCGF was additionally indicated by the responsiveness of this cell line to BCGF purified by a series of chromatographic steps. The BCGF to which B A3 cells responded had a m.w. of 14,000 and was similar to low m.w. BCGF reported from other laboratories. Surface characterization of B-A3 cells, analyzed by flow cytometry with a panel of monoclonal antibodies, demonstrated that the majority of B-A3 cells were stained positively with Leu-12, HLA-DR, and surface IgG markers, whereas staining with surface IgM, IgD markers, pan T cell markers (Leu 4 and Leu-9), and IL 2 receptor (Tac) were consistently negative. Taken together, the human lymphoblastoid cell line we present here responded specifically to a low m.w. BCGF. This cell line may be of value in the purification of BCGF to homogeneity, in studies of the interactions of BCGF with human B cells, and in the identification of the BCGF receptor. PMID- 3031162 TI - HTLV-III large envelope protein (gp120) suppresses PHA-induced lymphocyte blastogenesis. AB - The addition of inactivated preparations of purified human T cell lymphotropic virus (HTLV-III) was found to inhibit normal human lymphocyte phytohemagglutinin (PHA)-induced blastogenesis but had no effect on concanavalin A (Con A), pokeweek mitogen, or allogeneic stimulation. The inhibition was concentration-dependent and also dependent on adding the virus preparation before or at the same time as PHA. The CD4 molecule is the receptor for HTLV-III binding. Immunopurified large envelope protein (gp120) from HTLV-III was found to bind to the CD4 molecule and also inhibited PHA-induced lymphocyte blastogenesis. These results suggest that the gp120 viral protein may alter immune function by binding to the CD4 molecule, which in turn serves as an "off" signal to lymphocyte response to PHA stimulation. Alternatively, by binding to the CD4 molecule, gp120 may interfere with the interaction of this molecule with class II histocompatibility antigens on accessory cells, thus selectively suppressing PHA response. PMID- 3031161 TI - LTB4 induced activation signals and responses in neutrophils are short-lived compared to formylpeptide. AB - Leukotriene B4 (LTB4) was shown to be a potent stimulator of neutrophil actin polymerization as detected by right-angle light scatter and rhodamine-phalloidin staining of F-actin. When we compared the kinetics of this neutrophil cytoskeletal response to the chemoattractants formylpeptide and LTB4, we observed that the response to LTB4 was markedly shorter-lived. To understand the basis of this result, we re-examined the kinetics of superoxide generation, elastase release, intracellular calcium elevation, and phosphoinositide metabolism in neutrophils stimulated with LTB4 and N-formylhexapeptide. LTB4 was relatively inefficient in producing superoxide and in releasing elastase. Although both responses were initiated with similar rapidity, they turned off sooner with LTB4 as compared with N-formylhexapeptide stimulation. Intracellular calcium elevation, a signal that is necessary for superoxide generation and degranulation, was of similar magnitude but of shorter duration in response to LTB4 as compared with the N-formylhexapeptide. The LTB4-induced rise of phosphatidic acid also was not sustained as long as the N-formylhexapeptide induced increase. Prior exposure of neutrophils to LTB4 did not inhibit subsequent stimulation of superoxide generation by N-formylhexapeptide. Thus, the inability of LTB4 to stimulate superoxide generation was not due to LTB4-induced global inhibitory signals. The deficiency in LTB4-induced superoxide and elastase responses may be related to short-lived LTB4-induced activation signals and/or the number of receptors contributing to these responses. PMID- 3031163 TI - Cytostatic and cytotoxic activity of tumor necrosis factor on human cancer cells. AB - The cytostatic and cytotoxic activity of human recombinant tumor necrosis factor (rTNF) was assayed on different tumor cell lines. Human BT-20 breast and ME-180 cervix cancer cells were growth-inhibited by rTNF, whereas two other cell lines were not significantly inhibited. However, when protein synthesis was inhibited by cycloheximide, rTNF was cytotoxic for these cells but not for BT-20 cells. This finding suggested that different mechanisms are responsible for the cytostatic and cytotoxic activity of rTNF. The sensitivity of different cell lines to rTNF could not be correlated with a high number or affinity of rTNF receptors. Occupancy of only a few receptors was sufficient for rTNA cytotoxicity, but an increase in receptor number after treatment with interferon gamma, or a decrease after pretreatment with cycloheximide, correspondingly enhanced or depressed the cytotoxicity of rTNF. It seemed possible that some cells could be protected from this effect of rTNF by synthesizing "protective" proteins. While searching for such proteins, we observed that rTNF induced the synthesis of two polypeptides in SK-MEL-109 melanoma cells, but not in other cancer cells. Actinomycin D added with rTNF abolished synthesis of these polypeptides, suggesting that rTNF induced transcription of the corresponding mRNAs. Surprisingly, rTNF stimulated growth of SK-MEL-109 cells cultured in medium with low serum. PMID- 3031164 TI - An improved ELISA for the detection of antibodies to ovine and caprine lentiviruses, employing monoclonal antibodies in a one-step assay. AB - An improved ELISA for the detection of antibodies to ovine and caprine lentiviruses has been developed. The assay employs two monoclonal antibodies (Mabs) directed against the major core protein (p28) of the virus in a modified double antibody sandwich blocking procedure. The modification was necessary to circumvent the consequences of using low affinity Mabs and to lower the chance of false-negative results. A novel one-step concept was developed in which washing between steps was largely omitted. The new assay was designated complex trapping blocking (CTB)-ELISA. A range of sera from sheep and goats was tested comparatively in CTB-ELISA, in an indirect ELISA and in an agar gel precipitation test. The CTB-ELISA proved sensitive and highly specific and in addition reliable and easy to perform. PMID- 3031167 TI - Cerebellar glioblastoma. PMID- 3031166 TI - Reduction of ferricytochrome C may underestimate superoxide production by monocytes. AB - Superoxide production by stimulated phagocytes is commonly measured by reduction of ferricytochrome C, with specificity of the assay assumed if the reaction is inhibited by superoxide dismutase (SOD). Most preparations of ferricytochrome C contain a small proportion in the reduced (ferro) form, and this is also formed by the reaction of ferricytochrome C with superoxide. The generation of other reactive oxygen intermediates, such as hydrogen peroxide or hydroxyl radical, could cause oxidation of ferrocytochrome C and consequent underestimation of superoxide production. In support of this, it has been demonstrated that exogenous catalase enhanced the reduction of ferricytochrome C by phorbol myristate acetate (PMA)-stimulated human monocytes. Control experiments confirmed that this was due to enhanced detection rather than increased production of superoxide. In addition, SOD was found to promote oxidation of ferrocytochrome C by PMA-stimulated human monocytes, but this was also inhibited by catalase. These effects of catalase and SOD on ferricytochrome C reduction/ferrocytochrome C oxidation were also demonstrated when superoxide was produced independently of monocytes by a xanthine and xanthine oxidase generating system. It is concluded that the assay of superoxide, using 'SOD inhibitable' reduction of ferricytochrome C, underestimates superoxide production. PMID- 3031165 TI - Partial purification of lymphoblasts after in vitro immunization increases the yield in Ig-producing hybridomas. AB - Using in vitro immunization with a human plasma protein (apolipoprotein-A1) as antigen, we have shown that it is possible to prepare more monoclonal antibodies using a ten-fold lower concentration of antigen compared to in vivo immunization procedures (Weech et al., 1985). In addition, we can increase the number of Ig producing hybridomas after in vitro immunization by a simple one-step separation of the lymphoblasts on a Percoll gradient before the fusion procedure. In order to apply this procedure to in vivo immunization techniques, it is necessary to expand the B-blast/plasma cell population by culturing the spleen cells for 4-6 days before fusion. Only antibodies of the IgM class were produced with the in vitro technique. However, by combining in vivo priming with in vitro immunization, it is possible to produce specific antibodies to both IgG and IgM classes. PMID- 3031168 TI - [Benign tumors of the posterior urethra in children. Apropos of an unusual case of rhabdomyoma of fetal type]. AB - The authors report a very unusual case of urethral tumor in a child: fetal rhabdomyoma, after nephroblastoma treated by enlarged nephrectomy. Excellent result is observed four years after endoscopic treatment of this prostatic urethral tumor. Clinical and histological features of posterior urethral tumors and rhabdomyoma are recalled; the association nephroblastoma-tumor of urethra is discussed. PMID- 3031169 TI - Cytomegalovirus infection and reinfection transmitted by heart transplantation. PMID- 3031170 TI - Effects of human immunodeficiency virus on the cellular immune response to Epstein-Barr virus in homosexual men: characterization of the cytotoxic response and lymphokine production. AB - We investigated Epstein-Barr virus (EBV)-specific T cell responses in homosexual men with, and at risk for, AIDS. We studied healthy laboratory workers, healthy homosexual men, and patients with AIDS-related complex or AIDS. The cytotoxic activity, absolute number of T4 lymphocytes, and interleukin-2 (IL-2) production decreased, whereas the relative number of Ia+ lymphocytes increased with the extent of infection with the human immunodeficiency virus (HIV). Cytotoxic activity correlated positively with the number of T4 lymphocytes (r = .56, P less than .001) and the amount of IL-2 produced (r = .47, P less than .01) but not with interferon production. Recombinant IL-2, but not gamma interferon, could restore cytotoxic T cell activity to control levels in patients with early HIV infection. EBV-specific serological studies paralleled the T lymphocyte investigations. The increased EBV activity observed in progressive HIV infection may be related to a diminution in the auto-reactive population of the T4 lymphocyte subset and may be amenable to IL-2 reconstitution. PMID- 3031171 TI - Interaction of strain AD169 and a clinical isolate of cytomegalovirus with peripheral monocytes: the effect of lipopolysaccharide stimulation. AB - The effect of human cytomegalovirus (CMV) infection on lipopolysaccharide (LPS) stimulated and unstimulated monocytes from seronegative donors was studied by using the laboratory-adapted strain AD169 and a recent clinical isolate. LPS stimulated and unstimulated monocytes infected with the isolate showed expression of immediate-early CMV antigens and were significantly more suppressive for lymphocyte proliferation than were strain AD169-infected monocytes, which rarely expressed detectable viral protein. Human CMV infection of LPS-stimulated and unstimulated monocytes resulted in abrogation of interleukin-1 activity, with the effect being marked in LPS-stimulated monocytes infected with the clinical isolate of CMV. Addition of interleukin-1 to infected, stimulated monocytes completely restored lymphoproliferative responses to phytohemagglutinin, whereas addition of this leukokine to infected, unstimulated cells could not restore this response. PMID- 3031172 TI - Rapid clearance of cytomegalovirus-specific IgG after repeated intravenous infusions of human immunoglobulin into allogeneic bone marrow transplant recipients. AB - We studied the kinetics of the disappearance of CMV-specific IgG from the serum of 18 allogeneic bone marrow transplant recipients who were receiving repeated intravenous infusions of immunoglobulin. Peak serum titers occurred 24-48 hr after infusion. The mean half-life of IgG to CMV varied from 30 to 70 hr. Additional studies showed that this unexpectedly short half-life was not specific for either the preparation used or the type of patient studied. Repeated treatment did not prevent patients from developing CMV infections (incidence, 50%); however, none of the patients developed CMV interstitial pneumonitis. Treatment had to be discontinued for one patient because of a serum sickness-like syndrome. Our results suggest that rational, time-sequential passive immunization regimens should be developed. PMID- 3031173 TI - Vaccination with recombinant herpes simplex virus glycoproteins: protection against initial and recurrent genital herpes. AB - The natural history of initial and subsequent recurrent herpes simplex virus (HSV) genital infection was studied in guinea pigs immunized with HSV glycoprotein vaccines before vaginal challenge with HSV-2. The vaccines used included HSV-1 glycoprotein B and HSV-1 glycoprotein D, both prepared by recombinant DNA techniques, and mixtures of HSV-1 or HSV-2 glycoproteins, prepared from infected cell monolayers by lectin chromatography. Immunizing guinea pigs with subunit HSV glycoprotein vaccines favorably altered the clinical and virological course of initial infection and substantially reduced the incidence, frequency, and duration of recurrent herpetic infections. The extent of protection was influenced by the nature of the glycoprotein vaccine, the immunizing dose, and the co-administration of adjuvant. PMID- 3031174 TI - Immunization of experimental animals with reconstituted glycoprotein mixtures of herpes simplex virus 1 and 2: protection against challenge with virulent virus. AB - Artificial mixtures of the glycoproteins B, C, D, and E of herpes simplex virus 1 and 2 (HSV-1 and HSV-2), purified individually from infected Vero cell lysates by immunoaffinity to monoclonal antibodies, were bound to an aluminum hydroxide gel and were used to immunize mice, guinea pigs, and owl monkeys (Aotus trivirgatus) once or twice (mice and guinea pigs) to as many as four times (owl monkeys). In all animals tested, low levels of neutralizing antibodies were detected only after two or more immunizations. Lymphocyte transformation tests in owl monkeys suggested low or borderline levels of cellular immunity. The survival of immunized mice after intracerebral challenge was inversely related to the challenge dose. Immunized guinea pigs challenged by intravaginal inoculation showed reduced morbidity at the site of inoculation and were protected from CNS disease. Both immunized and nonimmunized monkeys were highly susceptible and could not be differentiated with respect to morbidity or mortality when challenged with 1,000 pfu of HSV-2 by the intravaginal route. PMID- 3031175 TI - Aseptic meningitis due to varicella-zoster virus: serum antibody levels and local synthesis of specific IgG, IgM, and IgA. AB - We used an indirect enzyme immunoassay to describe the evolution of serum levels and the intrathecal production of IgG, IgA, and IgM antibodies to varicella zoster virus (VZV) in eight patients with a syndrome of acute aseptic meningitis (AAM) and evidence of intrathecal production of VZV-specific IgG antibodies. Four of the eight patients showed no cutaneous zoster while hospitalized. Our results suggested an etiologic relation between VZV and AAM in all cases. Furthermore, we observed some differences in the pattern of evolution of antibodies in serum and cerebrospinal fluid in relation to the presence or absence of cutaneous lesions in our patients. These differences could reflect different pathogenic mechanisms in the spread of VZV to the central nervous system and in the production of the AAM syndrome. PMID- 3031176 TI - An in vitro model of the wound microenvironment: local phagocytic cell abnormalities associated with in situ complement activation. AB - An in vitro model was developed to investigate the inflammatory response to tissue damage. Human fibroblasts were heat killed and incubated with serum. Complement studies showed activation of the alternative pathway proportional to the number of dead cells; C3 was fixed on dead cells, and C5a was generated. Neutrophils (PMNLs) adhered to killed fibroblasts, a process requiring fresh serum. After adhering to killed fibroblasts in the presence of serum, PMNLs exhibited depressed chemotactic responsiveness to activated serum and reduced bactericidal activity against preopsonized Staphylococcus aureus. These data suggest that thermally killed cells activate and fix complement, a process generating cleavage products that, in turn, recruit PMNLs and bind them to the inflammatory site. Thus, in our model, dead tissue activates humoral mechanisms and inflammatory cells; this process results in depressed in situ host-defense function upon subsequent local challenge with microbes. PMID- 3031178 TI - Glomangiosarcoma with metastasis to the maxilla. AB - A thorough review of the literature disclosed very little information on glomangiosarcoma with no reported cases of verified metastases. We present a case which may be the first instance of oral metastatic glomangiosarcoma to be reported in the literature. PMID- 3031177 TI - Granulocyte colony-stimulating factor and differentiation-induction in myeloid leukemic cells. AB - The granulocyte colony-stimulating factor (G-CSF) belongs to a family of hemopoietic growth factors regulating the production of granulocytes and macrophages. Murine G-CSF stimulates the proliferation and differentiation of precursors of neutrophilic granulocytes and is also able to stimulate the functional activities of mature neutrophils. Among the hemopoietic growth factors, G-CSF has an outstanding capacity to induce terminal differentiation and suppression of self-renewal in myeloid leukemic cells. Murine and human G-CSF's show complete biological cross-reactivity across species and bind equally well to G-CSF receptors of either species. Specific receptors for G-CSF exist on all normal neutrophilic cells and have not been lost in the generation of primary human myeloid leukemias. This data indicates that G-CSF may be a useful reagent in the treatment of myeloid leukemia, in hemopoietic regeneration and in increasing resistance against infections. PMID- 3031179 TI - Benign neurogenic tumors of the oral cavity. AB - Neurogenic tumors are rare in the oral cavity, particularly so when malignant. Traumatic neuroma, although usually included with neurogenic tumors, is a reactive process rather than a true neoplasm. Neurofibroma and schwannoma derive from nerve fibers, the perineurium, the endoneurium and the neurolemmomal cells. They present histological differences. The neurofibroma may present in solitary and generalized types; the latter also known as neurofibromatosis or von Recklinghausen's disease of the skin. We here report typical cases of benign neurogenic tumors of the oral cavity. PMID- 3031180 TI - [Immunization of healthy children with live attenuated varicella vaccine (OKA strain)]. PMID- 3031181 TI - [Malignant fibrous histiocytomas arising in the thoracic area--a clinical and immunohistochemical study]. PMID- 3031182 TI - Krukenberg tumors of the ovary: a clinicopathologic analysis of 112 cases. AB - The records from 1965 to 1985 in the Department of Obstetrics and Gynecology Kurume University Hospital were reviewed. In this period, 112 cases were fulfilled the criteria of Krukenberg tumor and they were carefully screened to clinical history, physical findings, operative findings, and histologic character of the lesion. The incidence of Krukenberg tumors in all ovarian cancer was 17.8%. The average age was 45.4 years with ranged from 23 to 71. As expected from this age distribution, 73 patients (65%) were 50 years of age or younger. A primary carcinoma of the stomach was found in 78 (69.6%) of 112 patients. In three cases, the Krukenberg tumors were associated with a recent pregnancy. Two cases were associated with ovarian dermoid cyst. Microscopically, both ovaries were involved in all cases that could be evaluated for bilaterality. The relation between age distribution and histological characteristics was examined. Signet ring type carcinomas were observed frequently in young women and gland-formation type carcinomas in older women. Stromal hyperplasia of the tumors were remarkably common in younger women. In the patients with a recent pregnancy, the abdominal mass developed rapidly after delivery. In addition, estrogen receptors were present in all experimented cases. The above results suggested that estrogen influences tumor development and the histological character many of Krukenberg tumor. PMID- 3031183 TI - Dapsone-induced neuropathy compounds Hansen's disease nerve damage: an electrophysiological study in tuberculoid patients. AB - In 17 previous cases of dermatological disorders, an axonal motor neuropathy was described as a dapsone (DDS) therapy side effect. In this study, we attempted to assess DDS-induced neuropathy in the ulnar and popliteal nerves of 39 tuberculoid Hansen's disease patients using electrophysiological recordings at the time of DDS withdrawal, owing to dermatological improvement, and 4 months after. Distal motor latencies, conduction velocities at forearm and leg and above the epicondyle and the neck of the fibula were improved at a highly significant level. Twenty-five percent of the patients presented abnormal values (outside of the 95% confidence interval) at the first recording session compared to those at the second session. By contrast, parameters exploring the degree of innervation of distal muscles showed a progressive denervation. These results lead to an impairment of Hansen's disease neuropathy during DDS therapy affecting the motor conduction velocities of one quarter of the patients, and are discussed in terms of physiopathological mechanisms. PMID- 3031184 TI - Dapsone neuropathy--report of three cases and pathologic features of a motor nerve. AB - We report the clinical features, electrophysiologic findings, and dapsone and isoniazid excretion studies in three young people who ingested excessive amounts (2-4 times the prescribed dose) of dapsone for hypopigmented macules and who developed, subacutely, progressive motor neuropathy a few months later. Pathologic studies on a biopsied motor nerve confirmed the electrophysiologic conclusion of distal motor axonopathy. All made a rapid recovery in a few months after dapsone was stopped, although electrical abnormalities persisted. One patient was a rapid acetylator of isoniazid. PMID- 3031186 TI - [A case of primary hepatocellular carcinoma with hypouricemia]. PMID- 3031185 TI - [Use of calcium phosphate ceramics in periodontal therapy. 6. Clinical application of hydroxyapatite granules]. PMID- 3031187 TI - [A case of multiple myeloma with hepatocellular carcinoma]. PMID- 3031188 TI - [Intracellular responses of trigeminal sensory neurons to peripheral and central stimulation]. PMID- 3031189 TI - Receptor binding and activation of polymorphonuclear neutrophils by tumor necrosis factor-alpha. AB - The interaction of highly purified recombinant human tumor necrosis factor-alpha (rTNF-alpha) with human polymorphonuclear neutrophils (PMNs) was investigated. Binding of 125I-rTNF-alpha to PMN reached maximum levels in 30 min at 37 degrees C and in 2 h at 4 degrees C. Scatchard analysis of competitive binding data indicated approximately 6000 receptor sites per cell and a Kd of 1.37 nM. Binding data at 37 degrees C indicated a rapid internalization of rTNF-alpha. Following this receptor-mediated interaction, recombinant TNF-alpha was found to inhibit the migration of PMNs under agarose and to enhance PMN production of superoxide anion (O-2) in a dose-dependent manner. Furthermore, rTNF-alpha-activated PMNs caused a marked disruption of human umbilical-vein-derived endothelial cell monolayers and caused inhibition of their proliferative activities. These data substantiate the role of TNF-alpha as an activator of PMN functions and indicate that PMN/TNF-alpha/endothelial cell interactions may play a major role in inflammatory reactions. PMID- 3031190 TI - L-fucose, D-mannose, L-galactose, and their BSA conjugates stimulate macrophage migration. AB - The effect of selected monosaccharides on the random migration of normal adult rabbit alveolar macrophages (AM) was investigated. It was observed that 10 mM of L-fucose, L-galactose, or D-mannose stimulated AM migration 1.5-2.0 times. In addition, derivatives of L-fucose and D-mannose occupying the carbon-6 position such as L-fucosyl-lactose, D-mannose-6-phosphate, D-mannitol, and mannan enhanced the migration of AM, whereas derivatives of L-fucose and D-mannose in the carbon 1 position produced no migration enhancement. Macrophage migration enhancement activity that was produced spontaneously by spleen cell cultures from normal young rabbits was destroyed by treatment with L-fucosidase. Accordingly, the migration enhancement factor (MEF) found in spleen cell culture supernatants appeared to depend on L-fucose conjugated to some protein carrier because MEF was non-dialyzable. When normal adult AM were treated with L-fucosidase, they lost their responsiveness to migration inhibitory factor (MIF) but retained their responsiveness to MEF. We have interpreted this to mean that the MIF and MEF receptors are distinct. Synthetic MEFs were prepared by conjugating L-fucose, D mannose, of L-galactose to bovine serum albumin (BSA). It was noted that these sugar-BSA conjugates were about 200 times more effective than the corresponding free sugars in producing migration enhancement. In addition, these sugar-BSA conjugates neutralized MIF activity in a migration inhibition test. PMID- 3031191 TI - Chromatographic characteristics of pro-opiomelanocortin-derived peptides from the rat transplantable tumour 7315a. AB - Adrenocorticotrophin (ACTH) and other pro-opiomelanocortin (POMC)-derived peptides produced by the 7315a corticomammotrophic tumour have been poorly studied although they elicit profound hypertrophy and hyperplasia in the adrenal glands of recipient Buffalo rats. Tumour extracts were chromatographed on Sephadex G-75 and fractions monitored for POMC-derived peptides by four radioimmunoassay (RIA) systems: ACTH, alpha-MSH, beta-lipotrophin (beta LPH)/endorphin and N-terminal POMC (N-POMC). Chromatograms were compared with those of pars distalis extracts from normal Buffalo rats. All four RIA systems detected immunoreactive material in tumour extracts. ACTH, beta-LPH/endorphin and N-POMC were present in approximately equimolar amounts (ACTH content 93.40 +/- 5.27 (S.E.M.) pmol/g) whereas alpha-MSH was present in smaller amounts (2.83 +/- 0.13 pmol/g). Total peptide content correlated well with tumour weight. ACTH immunoreactivity (IR) in Sephadex chromatograms was located in a large 20,000 mol. wt peak, an ACTH(1-39) peak and a smaller peak coinciding with ACTH(1-24). The latter two peaks showed biological activity consistent with ACTH(1-39) and an ACTH (1-24)-like peptide respectively. The beta-LPH/endorphin RIA revealed a peak eluting at approximately 20,000 mol. wt which could not be ascribed to any known POMC peptide containing the endorphin sequence. A beta-LPH-like peak, a beta endorphin-like peak and a smaller-sized peak, which contained the bulk of the beta-LPH/endorphin IR, were detected; the low molecular weight peak probably representing alpha- or gamma-endorphin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3031192 TI - Response of adrenal cells to parathyroid hormone stimulation. AB - Dispersed adrenal cells of avian and bovine origin were incubated with human (h) 1-34 parathyroid hormone (PTH(1-34)), bovine (b) PTH(1-84), bPTH(3-34), avian (a) PTH, and with ACTH. Kidney tubular cells, the established target cells for PTH, were used for comparison. Cyclic AMP (cAMP) accumulation and steroid hormone (aldosterone and corticosterone) secretion were measured in response to the hormones. In the bovine adrenocortical cells PTH of both bovine and avian origin stimulated cAMP production, but the aPTH action was more pronounced. The maximal cAMP response of avian adrenocortical cells to aPTH was 15-fold greater than that of ACTH, but a 20-times higher concentration of aPTH was required to reach half maximal response. Avian PTH stimulated steroid hormone secretion in the chick adrenocortical cells, but the induced secretion was similar to that induced by ACTH, despite the difference in cAMP accumulation. Human PTH(1-34) and bPTH(1 84), which stimulated cAMP production in kidney cells, and the conventional antagonist bPTH(3-34) inhibited aPTH stimulation of cAMP accumulation in avian adrenocortical cells, but did not interfere with ACTH action. Furthermore, cAMP stimulation by aPTH and ACTH in avian adrenal cells was additive. The results establish the adrenal as a target organ for PTH, and suggest that the PTH acts through specific receptors, distinct from those for ACTH. PMID- 3031193 TI - Regulation of ecdysone biosynthesis in the tobacco hornworm, Manduca sexta: titre of the haemolymph stimulatory factor during the last larval instar. AB - A recently isolated haemolymph protein appears to be an important regulator of ecdysone biosynthesis by prothoracic glands in Manduca sexta. Using a dose response titration protocol, the haemolymph titre of this stimulatory factor was determined during the last larval instar. The titre was high (greater than 2.0 U ml-1) on days 0 and 1, then dropped significantly to 0.55 U ml-1 on day 2, and remained depressed until day 4. The titre of the stimulatory factor then increased to a peak of 1.62 U ml-1 on day 7, and remained elevated (approx. 1.1 U ml-1) until the end of the instar. A set of physical and biochemical criteria was used to confirm that the stimulatory activity present in haemolymph on different days of the instar represented the presence of the factor. The data are consistent with the hypothesis that fluctuations in the titre of the haemolymph stimulatory factor play a critical role in regulating ecdysone biosynthesis during larval-pupal development. PMID- 3031194 TI - Suppression of sensory to motor synaptic transmission and narrowing of the sensory neurone action potential by arginine vasotocin in Aplysia californica. AB - The vertebrate neurohypophysial peptide arginine vasotocin (AVT), which may be endogenous to the Aplysia central nervous system, was tested for its effect on sensory to motor neurone synaptic transmission. In the semi-intact preparation, superfusion of AVT (10(-6) moll-1) over the abdominal ganglion decreased the amplitude of both the gill withdrawal reflex and the short-latency excitatory postsynaptic potentials (EPSPs) evoked in gill and siphon motor neurones by single action potentials elicited in sensory neurones. AVT slowed the rate of rise of the EPSP, enhanced the rate of homosynaptic depression, and reversibly decreased the duration of the action potential of mechanosensory neurones in isolated, perfused abdominal and pleural ganglia. Frequency-dependent prolongation of action potentials of pleural sensory cells was also decreased by application of AVT. Because this peptide has been shown to modulate the gill withdrawal reflex and its subsequent habituation, the hypothesis that AVT plays a physiological role in the expression of the suppressed behavioural state is proposed. In addition, it is proposed that modulation of the reflex by AVT occurs in part by shortening the duration of the sensory neurone action potential. PMID- 3031195 TI - Internalization of interleukin 2 is mediated by the beta chain of the high affinity interleukin 2 receptor. AB - High-affinity IL-2-R correspond to a membrane receptor complex composed of two different IL-2-binding proteins, the Tac antigen (alpha chain) and a 70-75 kD beta chain. Using cell lines that express either the alpha or the beta protein, we demonstrate that IL-2 internalization occurs when ligand is bound to the isolated beta chain, but not when it is bound to the isolated alpha chain. The kinetics of IL-2 internalization mediated by the intermediate-affinity beta chain were nearly identical to those of the high-affinity alpha/beta heterodimer (t1/2 of 10-15 min), and each type of receptor targeted the bound IL-2 for intracellular degradation in lysosomes. The beta chain thus appeared to provide the essential element necessary for ligand internalization by both types of IL-2 R. PMID- 3031196 TI - Induction and inhibition of in vitro oocyte maturation and production of steroids in fish follicles by forskolin. AB - The effects of forskolin (FK) on in vitro oocyte maturation and production of steroids were examined in Oryzias latipes. When oocytes within preovulatory follicles were preincubated in the presence of FK for 2-10 hr, they matured normally after additional incubation for 10-20 hr in plain culture medium. Naked (follicle cell-free) oocytes did not mature under these conditions. FK stimulated dose-dependent production of steroids (estradiol-17 beta, E2, and 17 alpha,20 beta-dihydroxy-4-pregnen-3-one, 17 alpha,20 beta-diOHprog) and cAMP in follicle (granulosa) cells. On the other hand, exposure to FK within 2 hr after 17 alpha,20 beta-diOH prog stimulation caused reversible inhibition of gonadotropin (PMS)- or 17 alpha,20 beta-diOH prog-induced maturation of the intrafollicular oocytes in vitro. FK also significantly inhibited the 17 alpha,20 beta-diOHprog induced maturation of naked oocytes, suggesting the existence of adenylate cyclase in fish oocytes. These data indicate that in Oryzias latipes, FK induces oocyte maturation by stimulating follicular production of maturation-inducing steroid (MIS), probably 17 alpha,20 beta-diOH prog, via an increase in cAMP, and that it may inhibit oocyte maturation by increasing ooplasmic cAMP and some inhibitory interaction between the granulosa cells and the oocyte through intercellular communication. PMID- 3031197 TI - Effects of catecholamines on early development of the chick embryo: relationship to effects of calcium and cAMP. AB - Catecholamines (dopamine or norepinephrine) injected under the blastoderm of the unincubated chick embryo produced a thickened primitive streak and prevented the migration of axial mesoblast after 24 h. The mesoblastic cells that accumulated in the primitive streak contained many intracytoplasmic yolk granules. After 48 h, neural tube, notochord, and somites were severely affected, and their cells appeared loaded with yolk inclusions. Heart, lateral plates, blood cells, and blood vessels differentiated normally. At the onset of gastrulation, the level of glycogen was fivefold lower in catecholamine-treated embryos than in control embryos. Injection of glucose plus dopamine, at equimolar concentrations resulted in normal development both at 24 h and at 48 h. Because adrenergic stimulation of glycogenolysis in differentiated cells is usually mediated by cAMP and/or by calcium, we attempted to determine whether these substances could reproduce the effects of catecholamines. Only calcium was able to produce, to a limited extent, the same morphogenetic disturbances as those produced by catecholamines, whereas the chelating agent, ethylenediamine tetraacetic acid, when administered with dopamine, partially inhibited the effects of catecholamines. An increase in the number of yolk granules was the only common finding among embryos treated with cAMP and catecholamines. Blood and a well differentiated, gastrular endoderm always developed, independently of the nature of the substance with which the embryos had been treated. Morphogenetic disturbances caused by exogenous catecholamines could be due to depletion of glucose. Alternatively, a different metabolic commitment might exist within the diverse populations of cells that constitute the mesoblastic layer. PMID- 3031198 TI - Effects of agar and Ficus awkeotsang makino on mucin degradation and intestinal permeability of phenol red in rats. PMID- 3031200 TI - Detection of serum antibody responses to RIT 4237 rotavirus vaccine by ELISA and neutralization assays. AB - Immunoglobulin class-specific determination by ELISA and a virus neutralization test (NT) were compared for the detection of the serum antibody responses after vaccination of infants with the RIT 4237 rotavirus vaccine (of bovine origin). A capture method was applied for specific IgM determination by ELISA because of its greater sensitivity and reproducibility over the conventional ELISA-IgM test. NT was improved through the use of an enzyme-labelled antibody for the detection of non-neutralized virus. In 6-12-month-old children, the most sensitive single test to detect an antibody response was the ELISA-IgM capture method, but the combination of ELISA-IgM and IgG tests or ELISA-IgM and NT detected the highest seroconversion rate of 79%. In newborn infants high levels of maternal antibody made the ELISA-IgG test unsuitable. ELISA-IgM gave a response rate of 31%, but NT with homologous virus was the most sensitive indicator of a serological response to the vaccine in this age group, yielding a 45% response rate. PMID- 3031201 TI - Intracerebral infection of Cebus apella with the XJ-Clone 3 strain of Junin virus. AB - To assess the usefulness of the South American primate Cebus apella as a model for neurovirulence of Junin virus, eight monkeys were inoculated with 10(5) LD50 of the attenuated XJ-Clone 3 Junin virus strain by the intrathalamic route. After the second week, weight loss and polyadenopathies were observed in most animals, one-half of which had a transient leukothrombocytopenia. Moderate clinical central nervous system (CNS) involvement was present in four of eight monkeys, while the rest had only mild neurologic signs. All recovered except one, which developed a deep coma and was killed in a pre-mortem stage at 18 days post infection (pi). Junin virus was isolated from the throat from five, from the blood from three, and from the brain from two monkeys. In the most severely ill animal, virus titers higher than viremia were detected in both inoculated and contralateral brain hemispheres, as well as in lung, lymph node, and small intestine. Junin antigens and "in vivo" bound immunoglobulins were detected by immunofluorescence (IF) in the brain of four animals at 18, 21, 40, and 155 days pi. Moderate lymphocytic parenchymal and meningeal infiltration were observed in the brain of four animals, and gliosis was also present in the most affected monkey. Although the clinical response to infection was not uniform, all infected monkeys developed high IF antibodies. Cebus apella cannot be used as a highly sensitive model for Argentine hemorrhagic fever (AHF). However, the results obtained show that the XJ-Clone 3 strain can replicate in the primate CNS and to induce lesions and immunoglobulin deposition. In addition, viral persistence is suggested by the late detection of viral antigens in brain at 40 and 155 days pi. PMID- 3031199 TI - Na+- and cGMP-induced Ca2+ fluxes in frog rod photoreceptors. AB - We have examined the Ca2+ content and pathways of Ca2+ transport in frog rod outer segments using the Ca2+-indicating dye arsenazo III. The experiments employed suspensions of outer segments of truncated, but physiologically functional, frog rods (OS-IS), intact isolated outer segments (intact OS), and leaky outer segments (leaky OS with a plasma membrane leaky to small solutes, but with sealed disk membranes). We observed the following. Intact OS or OS-IS isolated and purified in Percoll-Ringer's solution contained an average of 2.2 mM total Ca2+, while leaky OS contained 2.0 mM total Ca2+. This suggests that most of the Ca2+ in OS-IS is contained inside OS disks. Phosphodiesterase inhibitors increased the Ca2+ content to approximately 4.2 mM in intact OS or OS-IS, whereas the Ca2+ content of leaky OS was not altered. Na-Ca exchange was the dominant pathway for Ca2+ efflux in both intact and leaky OS/OS-IS. The rate of Na-Ca exchange in intact OS/OS-IS was half-maximal between 30 and 50 mM Na+; at 50 mM Na+, this amounted to 5.8 X 10(7) Ca2+/OS X s or 0.05 mM total Ca2+/s. This is much larger than the Ca2+ component of the dark current. Other alkali cations could not replace Na+ in Na-Ca exchange in either OS-IS or leaky OS. They inhibited the rate of Na-Ca exchange (K greater than or equal to Rb greater than Cs greater than or equal to Li greater than TMA) and, as the inhibition became greater, a delay developed in the onset of Na-Ca exchange. The inhibition of Na Ca exchange by alkali cations correlates with the prolonged duration of the photoresponse induced by these cations (Hodgkin, A. L., P. A. McNaughton, and B. J. Nunn. 1985. Journal of Physiology. 358:447-468). In addition to Na-Ca exchange, disk membranes in leaky OS showed a second pathway of Ca2+ transport activated by cyclic GMP (cGMP). The cGMP-activated pathway required the presence of alkali cations and had a maximal rate of 9.7 X 10(6) Ca2+/OS X s. cGMP caused the release of only 30% of the total Ca2+ from leaky OS. The rate of Na-Ca exchange in leaky OS amounted to 1.9 X 10(7) Ca2+/OS X s.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3031203 TI - Infection of a poikilothermic cell line (XL-2) with eastern equine encephalitis and western equine encephalitis viruses. AB - Eastern Equine Encephalitis (EEE) was in Cuba before the 1940s; the virus has been isolated from horses, birds, and rodents during epizootic as well as interepizootic periods. The only isolation of Western Equine Encephalitis (WEE) virus was from a sick pigeon found in the vicinity of Havana University. Both viruses can cause human disease; the isolation of WEE virus from the centre of an urban area emphasises the need for the prompt isolation and rapid identification of these agents. The object of this work was to compare the sensitivity of a continuous cell line (XL-2) from the toad, Xenopus laevis, with primary chick embryo cell cultures (CEC) routinely used for isolation as well as assay of these viruses. Both cell systems were infected with EEE virus isolated from horse brain and WEE virus isolated from a sick pigeon. A clear cytopathic effect (CPE) consisting of rounding and detachment of cells was observed in both cell cultures infected with the two viruses. By 18 hours post-infection, there was partial destruction of the cell monolayer and by 24 hours the CPE was total. The infectious titre of EEE and WEE viruses in XL-2 and CEC were similar. Both viruses produced small plaques (0.5-1.0 mm diameter) in XL-2 cells. Studies on the sensitivity of the XL-2 cells for direct isolation of the two viruses from field samples and for the detection of Cuban flaviviruses by the immunofluorescence test are in progress. PMID- 3031202 TI - Effect of combinations of difluoromethylornithine (DFMO) and 9[(1,3-dihydroxy-2 propoxy) methyl]guanine (DHPG) on human cytomegalovirus. AB - Both the nucleoside analog 9[(1,3-dihydroxy-2-propoxy)methyl]guanine (ganciclovir; DHPG) and the polyamine synthesis inhibitor difluoromethylornithine (DFMO) have been reported to inhibit replication of cytomegalovirus (CMV) in vitro. In these studies, DHPG inhibited human CMV replication at concentrations of 0.1 microM or greater, while DFMO was active (and then only inconsistently) at 5 mM or greater. (DFMO was added to cells 3 days before virus to maximize polyamine depletion.) However, DHPG combined with DFMO synergistically inhibited the replication of CMV strain AD169 and one wild-type CMV strain in human foreskin fibroblasts; the effect was evident on both virus yield and plaque formation. The synergistic effect of these two drugs on CMV replication could potentially be exploited clinically to limit drug toxicity or increase efficacy. PMID- 3031204 TI - Cloning and characterization of BK virus-related DNA sequences from normal and neoplastic human tissues. AB - DNA homologous to that of the known papovarius BK was cloned from the high molecular weight DNA of two human tissues: a normal liver and a kidney carcinoma. The clone isolated from human liver consisted of DNA indistinguishable from prototype BK by restriction enzyme analysis that used ten different enzymes. The DNA cloned from the human carcinoma of the kidney was subject to rearrangement in recombination-deficient bacteria, and exhibited a deletion of a small segment of DNA localized to the BK late region. Restriction fragments representing the BK origin and promoter regions are overrepresented in the tumor-derived clone. The possible significance of retrieval of defective viral genomes from tumor tissues is discussed. PMID- 3031205 TI - Unilateral tonsillar swelling in adults. PMID- 3031206 TI - Signet-ring cell carcinoma of the urinary bladder. PMID- 3031207 TI - Mania as a manifestation of end-stage renal disease. AB - A patient presented with mania as the primary psychiatric manifestation of end stage renal disease. Previous descriptions of mania secondary to renal failure have been reported only in the 19th century literature. This case demonstrates that mania may arise due to endstage renal disease and that it should be included in the differential of secondary mania. Implications of this case on the pathophysiology of mania are discussed. PMID- 3031208 TI - Involvement of both opiate and catecholaminergic receptors of ventromedial hypothalamus in the locomotor stimulant action of thyrotropin-releasing hormone. AB - To explore the mode of the locomotor stimulant action of thyrotropin-releasing hormone (TRH), rats with or without administration of opiate or catecholaminergic receptor antagonists were infused with TRH through previously implanted hypothalamic cannula. Administration of TRH, but not the normal saline or TSH, into the ventromedial hypothalamus caused an enhancement in both the gross movements (including stimulation of forward locomotion, head and body rearing) and fine movements (including increased grooming and head swaying). The locomotor stimulant action provoked by TRH was antagonized by pretreatment of ventromedial hypothalamus with either an alpha-adrenergic receptor antagonist (yohimbine), a dopaminergic receptor antagonist (haloperidol) or an opiate receptor antagonist (naloxone), but not with a beta-adrenergic receptor antagonist (propranolol). The results indicate that all the adrenergic, dopaminergic and opiate receptors in the ventromedial hypothalamus are involved in the TRH-induced hyperactivity in the rat. PMID- 3031209 TI - Hemes and hemoproteins. 3. The reaction of microperoxidase-8 with cyanide: comparison with aquocobalamin and hemoproteins. AB - The monomeric heme octapeptide from cytochrome c, microperoxidase-8, (MP-8), coordinates CN- with log K = 7.55 +/- 0.04 at 25 degrees C in 20% (v/v) aqueous methanol. Log K values are independent of pH between 6 and 9. A spectrophotometric titration of cyanoMP-8 between pH 5.5 and 13.8 gave a single pKa greater than or equal to 13.5 ascribed to ionization of the proximal His ligand. A study of the kinetics of the reaction of MP-8 with cyanide between pH 5.5 and 12, at 25 degrees C and mu = 0.1, indicates that formation of cyanoMP-8 occurs via three routes: attack of CN- on Fe(III) (k1 = 6.0 +/- 0.3 X 10(5) M-1 sec-1); attack of HCN on Fe(III) (k2 = 4.8 +/- 2.0 X 10(3) M-1 sec-1), followed by deprotonation and isomerization to form the C-bound species; and displacement of OH- by CN- when the proximal His ligand is ionized (k5 = 1.8 +/- 0.1 X 10(5) M 1 sec-1). These results are compared with available data for the reaction of cyanide with aquocobalamin and with various hemoproteins. PMID- 3031210 TI - Ternary complexes of Cu(II) ion with cimetidine and L-alanine. AB - (Cu(CM)A)B(C6H5)4 and Cu(CM)A(OH) X H2O (CM = cimetidine and HA = L-alanine) were prepared and characterized by elemental analysis, TG-DTA, IR, and electronic spectral data and magnetic susceptibility measurements. The EPR spectrum of (Cu(CM)A)B(C6H5)4 shows a distorted octahedral environment for the Cu(II) ion. PMID- 3031211 TI - Effect of inositol hexakisphosphate on the EPR properties of the nitric oxide derivative of ferrous dromedary (Camelus dromedarius) hemoglobin. Evidence for two polyanion binding sites. AB - The effect of inositol hexakisphosphate on the EPR properties of the nitric oxide derivative of ferrous dromedary (Camelus dromedarius) hemoglobin has been investigated at 110 K. In the absence of inositol hexakisphosphate, the nitrosyl derivative of dromedary hemoglobin shows an EPR spectrum with a rhombic shape and a weak hyperfine splitting in the gz region, a feature that is generally taken as characteristic of the high-affinity state of tetrameric hemoproteins. On addition of 1 mole of inositol hexakisphosphate/tetramer, three new hyperfine lines (Az = 1.7 mT), centered at gz = 2.01, appear; this type of spectrum is indicative of the low-affinity state of hemoglobins. A further addition of inositol hexakisphosphate, corresponding to a 20-fold molar excess, completely reverses the polyphosphate-dependent transition, giving an EPR spectrum that is exactly superimposable to that observed in the absence of the allosteric effector, i.e., is typical of the high-affinity state of the macromolecule. Both in the absence and presence of inositol hexakisphosphate, the EPR spectra are virtually independent of pH in the range explored (from 5.5 to 7.5). These results, taken together with the behavior of the nitric oxide derivative of human hemoglobin, provide further evidence for the existance in dromedary hemoglobin of two polyanion binding sites that affect in an opposite way the conformational equilibrium of the macromolecule. PMID- 3031212 TI - Effects of extracellular choline concentration and K+ depolarization on choline kinase and choline acetyltransferase activities in superior cervical sympathetic ganglia excised from rats. AB - The activities of choline kinase (CK) and choline acetyltransferase (ChAT) were examined in vitro in superior cervical sympathetic ganglia (SCG) excised from rats following aerobic incubation for 1 h in a medium containing various choline concentrations, with and without application of a high KCl level (70 mM). Ganglionic CK activity was strongly inhibited (by approximately 75%) at low extracellular choline concentrations (1-5 microM) but rose as the choline concentration was raised to 10-50 microM in the incubation medium, then fell and rose again with further increases in choline concentration. A similar but moderate accelerative effect on ganglionic CK activity was also observed after addition of acetylcholine (ACh; 1 mM) without eserine. Whereas specific CK activity did not change significantly in axotomized SCG, in which the ratio of glial cells to neurons is greatly increased for a week after the operation., it was remarkably increased after denervation, in which the preganglionic cholinergic nerve terminals had degenerated. When either a high KCl level or hemicholinium-3 (HC-3; 50 microM) was added to the medium in the presence or absence of choline, ganglionic CK activity was markedly inhibited. On the other hand, ChAT activity in the SCG remained at a significantly high level during incubation with low choline concentrations (1-10 microM), but the enhanced enzyme activity became inhibited as the extracellular choline concentration was raised to 50-100 microM in the medium. Addition of HC-3 to the medium did not alter ganglionic ChAT activity at low choline concentrations. However, application of quinacrine (10 microM) considerably reduced ganglionic CK activity and also suppressed ChAT activity induced by high KCl levels.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3031213 TI - Calcium-dependent nerve growth factor-stimulated hydrolysis of phosphoinositides in PC12 cells. AB - The effect of nerve growth factor (NGF) on the hydrolysis of phosphoinositides in PC12 cells was examined. Addition of NGF to PC12 cells prelabeled with [3H] inositol resulted in an increase in the formation of labeled inositol trisphosphate ([3H]IP3), inositol bisphosphate ([3H]IP2), and inositol monophosphate ([3H]IP), an observation indicating that NGF stimulated hydrolysis of the polyphosphoinositides. The increase in these inositol phosphates was detected as early as 15 s after addition of NGF. In the presence of LiCl, the accumulation of [3H]IP was linear for at least 20 min. The NGF-stimulated accumulation of [3H]IP was dose-dependent with a Kact of 0.17 nM and was dependent on the presence of extracellular calcium. In a calcium-free buffer containing EGTA, the NGF-dependent increase in accumulation of [3H]IP was not seen, and the basal level of [3H]IP accumulation was lower than that observed in the presence of extracellular calcium. Lanthanum inhibited both the basal and NGF stimulated accumulation of [3H]IP, whereas the calcium ionophore A23187, in the absence of NGF, stimulated an accumulation of [3H]IP. The maximal accumulation of [3H]IP in the presence of A23187 was the same as that observed in the presence of NGF. Incubation of the cells with both A23187 and NGF resulted in an accumulation of [3H]IP that was not significantly different from the effect of either agent alone. These results suggest that NGF rapidly stimulates the hydrolysis of phosphoinositides in PC12 cells and that this NGF-stimulated hydrolysis of phosphoinositides occurs by a calcium-dependent mechanism. PMID- 3031214 TI - Solubilisation of the benzodiazepine receptor from rat cerebellum by the detergent n-octylpentaoxyethylene. AB - The detergent n-octylpentaoxyethylene is one in the series of tenside detergents developed for membrane solubilisation. We have used this detergent to solubilise benzodiazepine receptors from rat cerebellum. The soluble receptor has an affinity (KD) for [3H]flunitrazepam of 1.8 nM +/- 0.2, which is not significantly different from that observed in synaptic membranes. Under optimal conditions (0.6% wt/vol), the number of soluble flunitrazepam binding sites (Bmax) of 0.35 pmol/mg protein suggests an apparent solubilisation of 40% of sites from the membrane. However, the absence of the characteristic facilitation of [3H]flunitrazepam binding by gamma-aminobutyric acid (GABA), cartazolate, and pentobarbital in this soluble receptor preparation suggests that such a preparation is unlikely to be a useful preparation for further studies on the molecular mechanisms underlying GABAA receptor function. PMID- 3031215 TI - Enzymic synthesis of leukotriene B4 in guinea pig brain. AB - Leukotriene B4 [5(S), 12(R)-dihydroxy-6, 14-cis-8,10-trans-eicosatetraenoic acid] was obtained from endogenous arachidonic acid when slices of the guinea pig brain cortex were incubated with the calcium ionophore A 23187. Enzymes involved in its synthesis, arachidonate 5-lipoxygenase [arachidonic acid to 5(S)-hydroperoxy-6 trans-8,11,14-cis-eicosatetraenoic acid and subsequently to leukotriene A4] and leukotriene A4 hydrolase (leukotriene A4 to B4), were present in the cytosol fraction. Arachidonate 5-lipoxygenase was Ca2+-dependent, and was stimulated by ATP and the microsomal membrane, as was noted for the enzyme from mast cells. The lipid hydroperoxides stimulated 5-lipoxygenase by four- to sixfold. The leukotriene A4 hydrolase activity was rich in brain, and the specific activity (0.4 nmol/min/mg of protein) was much the same as that of guinea pig leukocytes. High activities of these enzymes were detected in the olfactory bulb, pituitary gland, hypothalamus, and cerebral cortex. Since leukotriene B4 is enzymically synthesized in the brain, possible roles related to neuronal functions or dysfunctions deserve to be examined. PMID- 3031216 TI - Identification of atrial natriuretic factor receptor of neuroblastoma N4TG1 cells: binding characteristics and photoaffinity labeling. AB - We have found specific receptors for atrial natriuretic factor (ANF) in cultured neuroblastoma cells (N4TG1) of peripheral ganglionic origin. Scatchard analysis of the displacement binding revealed noninteracting, single-class binding sites with a KD of 1 X 10(-10) M and a density (Bmax) of 110,000-150,000 sites/cell. The cell-bound 125I-ANF was displaced by unlabeled ANF in a dose-dependent manner. Hormones unrelated to ANF such as angiotensins, adrenocorticotropic hormone, or arginine vasopressin were ineffective in displacing the cell-bound radioactivity. Using azidobenzoyl-125I-ANF as a photoaffinity ligand, an ANF receptor with an apparent Mr of 138,000 was identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis and autoradiography. The addition of unlabeled ANF (1 microM) to the incubation medium completely abolished the labeling of this protein band, but atriopeptin I (1 microM) or angiotensins I, II, and III (each 1 microM) were not effective in inhibiting the affinity labeling. The treatment of the neuroblastoma cells with ANF stimulated intracellular cyclic GMP levels in a dose-dependent manner with an EC50 of 5 nM. ANF (1 X 10(-7) M) stimulated cyclic GMP accumulation in less than 5 min by 30-fold as compared to the controls. PMID- 3031217 TI - Calcium-dependent release of tyrosine in brain elicited by stimulation of neuropeptide receptors. AB - We sought to establish whether the endogenous opiate-receptor agonist Met enkephalin (m-ENK) selectively modulates the release of endogenous tyrosine (Tyr) from brain slices prepared from the corpus striatum (CS). Amino acids (AAs) released from slices of CS and, for comparison, cerebral cortex (Cx) were measured by HPLC. Incubation of slices with m-ENK (1-10 microM) increased the basal release of Tyr (up to 293% of control) from CS, but not Cx, whereas other nonneurotransmitter AAs, phenylalanine (Phe) and valine (Val), were unchanged. The release of the putative neurotransmitter AAs glutamate (Glu), taurine (Tau), and glycine (Gly) were similarly increased by 50-150% with m-ENK in slices of CS, but not Cx. The enhanced release of AAs by m-ENK was prevented by removal of extracellular Ca2+ or by preincubation with the opiate receptor antagonist naloxone. Neuronal depolarization by potassium (5-55 mM) in the presence of Ca2+ did not affect the release of Tyr, whereas release of neurotransmitter AAs such as gamma-aminobutyric acid (GABA) were markedly increased. The increase in basal Tyr release by m-ENK was not the result of a decreased uptake of Tyr. Relative to slices, the basal release of Tyr, Phe, and Val from a synaptosomal (P2) preparation of CS was small (8-51%) compared to that of GABA, Gly, Glu, and Tau (49-123%). Nonetheless, m-ENK (10 microM) markedly increased the release of Tyr (to 833%), but not Glu, Gly, and Tau from the P2 fraction.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3031218 TI - Molecular cloning of cDNA to mRNA for a cerebellar spot 35 protein. AB - The nucleotide sequence of mRNA for rat cerebellar spot 35 protein, a Ca-binding protein, was determined from recombinant complementary DNA (cDNA) clones. The sequence was composed of 1,714 base pairs (bp) which included the 783 bp of the complete coding region, the 130 bp of the 5'-noncoding region, and the 801 bp of the 3'-noncoding region containing a polyadenylation signal. In addition, a polyadenylic acid [poly(A)] tail was also found. Because the size of spot 35 mRNA was estimated to be about 1,900 bases by Northern blot analysis, the longest insert was verified to contain a nearly full-length cDNA sequence including the poly(A) tail. The amino acid sequence of the protein deduced from the nucleotide sequence contains 261 amino acids and at least five Ca-binding domains. There was a high homology in the amino acid sequences (79%) and the nucleotide sequences (77%) between spot 35 protein and chick intestinal Ca-binding protein (28K). PMID- 3031219 TI - Search for viral nucleic acid sequences in the post mortem brains of patients with schizophrenia and individuals who have committed suicide. AB - Molecular hybridisation has been used to screen several areas of post mortem brain from 20 patients with schizophrenia, 23 individuals who were suspected of having committed suicide and 21 control cases, for viral nucleic acids. Cloned probes were able to detect picogram levels of viral DNA and to quantify herpes simplex type 1 DNA from encephalitic brain, but no sequences hybridising to cytomegalovirus, varicella zoster virus, herpes simplex type I or JC or BK papovaviruses were detected in any of the experimental samples. These findings are discussed with reference to the viral hypothesis of the aetiology of psychiatric disease. PMID- 3031220 TI - The variable clinical manifestations of ulnar neuropathies at the elbow. AB - In twenty-five cases of ulnar neuropathy at the elbow, the involvement of the fibres from three sensory and to four motor branches were examined clinically and, where possible, electrophysiologically. Of the sensory fibres, those from the terminal digital nerves were most commonly involved. The fibres to the hand muscles were much more frequently involved than those to the forearm muscles. These findings suggest that in ulnar neuropathies at the elbow there is variable damage to the fascicles within the nerve. PMID- 3031221 TI - Neurotoxic effects of n-hexane on the human central nervous system: evoked potential abnormalities in n-hexane polyneuropathy. AB - An outbreak of n-hexane polyneuropathy as a result of industrial exposure occurred in printing factories in Taipei area from December 1983 to February 1985. Multimodality evoked potentials study was performed on 22 of the polyneuropathy cases, five of the subclinical cases, and seven of the unaffected workers. The absolute and interpeak latencies of patterned visual evoked potential (pVEP) in both the polyneuropathy and subclinical groups were longer than in the normal controls. The pVEP interpeak amplitude was also decreased in the polyneuropathy cases. Brainstem auditory evoked potentials (BAEP), showed no difference of wave I latency between factory workers and normal controls, but prolongation of the wave I-V interpeak latencies was noted, corresponding with the severity of the polyneuropathy. In somatosensory evoked potentials (SEPs), both the absolute latencies and central conduction time (CCT) were longer in subclinical and polyneuropathy cases than in the unaffected workers and normal controls. From this evoked potentials study, chronic toxic effects of n-hexane on the central nervous system were shown. PMID- 3031222 TI - X-linked bulbo-spinal neuronopathy: a family study of three patients. AB - The clinical features of two brothers and one nephew with X-linked recessive bulbo-spinal neuronopathy are described. The neurophysiological investigations and sural nerve biopsy, previously unreported, confirmed that both motor and sensory nerves are affected. Because of the genetic implications, the importance is stressed of recognising this disorder as a separate entity which should not be classified with the spinal muscular atrophies. PMID- 3031223 TI - Another venereal disease with frequent nervous system involvement: neuro-AIDS. PMID- 3031224 TI - WR-2721 and high-dose cisplatin: an active combination in the treatment of metastatic melanoma. AB - Cisplatin, alone or in combination with other chemotherapeutic agents, is relatively inactive against metastatic melanoma. Prior trials have demonstrated partial response (PR) rates of less than 10% with cisplatin alone. WR-2721 is an organic thiophosphate compound, which in the animal model, selectively protects normal tissues against the toxicity of cisplatin chemotherapy. During the course of a phase I trial of WR-2721 and cisplatin, objective PRs were noted in patients with far advanced metastatic melanoma. These observations led us to perform a phase II trial of WR-2721 and cisplatin. Thirty-six patients received 128 courses of WR-2721 before cisplatin (60 to 150 mg/m2). All patients had progressive disease before treatment. Objective PRs were observed in 19 of 36 evaluable patients (53%). Three additional patients had minor responses (MRs). PRs occurred in 53% of patients with prior chemotherapy (ten of 19). Sites of responding metastases were subcutaneous disease (15 of 19 patients), lymph nodes (16 of 21 patients), lung (four of ten patients), and liver (eight of 17 patients). The median duration of response was 4 months, with a mean of 4.5 months (range, 1 to 8 months). Transient nephrotoxicity was observed in less than 5% of courses. In all cases, renal function returned to normal within 1 to 2 weeks. Hematologic toxicity was mild and infrequent. Nine patients developed peripheral neuropathy following a median cisplatin dose of 670 mg/m2. Twenty patients experienced mild clinical hearing loss. These data suggest that WR-2721 may potentiate the antitumor activity of cisplatin in metastatic melanoma. PMID- 3031225 TI - The superiority of combination chemotherapy including etoposide based on in vivo cell cycle analysis in the treatment of extensive small-cell lung cancer: a randomized trial of 288 consecutive patients. AB - Two hundred and eighty-eight patients with extensive small-cell carcinoma of the lung (SCCL) were entered into a three-arm prospective randomized trial. The purpose was both to compare etoposide with methotrexate (MTX) in a combination chemotherapy regimen otherwise consisting of vincristine (VCR), lomustine (CCNU), and cyclophosphamide (CTX) and to evaluate a treatment design based on cell kinetic observations suggesting enhanced sensitivity to etoposide three to six days after administration of VCR, CCNU, and CTX. In all three treatment arms, VCR, CCNU, and CTX were administered on day 1 of a 28-day cycle. In arm A, MTX was administered on days 14 and 17, while in arm B, MTX was replaced by etoposide administered on days 14 through 17. In arm C, MTX was also replaced by etoposide, but administered on days 3 through 6. Overall survival was significantly longer for patients treated with "early" etoposide (arm C; median, 33 weeks) as compared with arm A (MTX; median, 23 weeks) (P less than .05), but not statistically different from "late" etoposide administration (arm B; median, 27 weeks). However, for patients with initial favorable performance status (0 + 1), a significantly longer survival was obtained for those treated with early etoposide (arm C. median, 51 weeks) as compared with patients in arm A (median, 32 weeks) and arm B (median, 36 weeks) (P less than .05). Two-year survival was obtained in six patients (7%) in arm C compared with three patients (3%) in arm B and none in arm A. The study confirmed that etoposide is an active drug in the treatment of SCCL and when combined with CTX, CCNU, and VCR, the cell kinetic approach of an early administration yields the best results. PMID- 3031227 TI - Hepatic arterial infusion of mitoxantrone in the treatment of primary hepatocellular carcinoma. AB - Twenty-three patients (16 male, seven female) with hepatocellular carcinoma (HCC) were treated by hepatic arterial infusion (HAI) of mitoxantrone every 4 weeks. At each treatment, a catheter was inserted percutaneously into the main hepatic artery via the femoral artery under image intensification. Treatment consisted of a 24-hour continuous HAI of mitoxantrone, 6 mg/m2/d X 3 (eight patients) or 10 mg/m2/d X 3 (14 patients) without heparin. Eight patients had only one infusion, nine patients four infusions, five patients three infusions, two patients two infusions, and one patient five infusions. A partial response was seen in six patients, with a median duration of 20 weeks (range, 18 to 38 weeks). Five patients achieved stable disease, with a median duration of 20 weeks (range, 11 to 42 weeks). The median survival of the overall group was 22 weeks. Survivals of responding, stable, and nonresponding patients were 32 weeks, 24 weeks, and 9 weeks, respectively. Complications of catheter placement included asymptomatic dissection of the hepatic artery (one patient), and asymptomatic thrombosis of the hepatic artery (five patients). Three patients experienced mild nausea and vomiting, and six patients had mild to moderate alopecia. Granulocytopenia was frequent at both dose schedules. The granulocyte nadir was greater than 1,000/microL in 34% of evaluable courses, 500 to 1,000/microL in 32%, and less than 500/microL in 34% of courses. Two patients developed neutropenia-associated fever. A platelet nadir below 100,000/microL was seen after only 10% of courses, and only two patients had platelets below 50,000/microL. Seven patients received doxorubicin after progression on mitoxantrone. Four received systemic doxorubicin, 50 mg/m2, and three HAI of doxorubicin, 25 mg/m2, for three days. Two patients achieved partial response (18 weeks and 32 weeks) to HAI doxorubicin. Mitoxantrone has activity in HCC and is well tolerated when administered by HAI. It is not entirely cross-resistant with doxorubicin. PMID- 3031226 TI - Multimodal therapy for limited small-cell lung cancer: a randomized study of induction combination chemotherapy with or without thoracic radiation in complete responders; and with wide-field versus reduced-field radiation in partial responders: a Southwest Oncology Group Study. AB - In 1979 we initiated a phase III study in the Southwest Oncology Group (SWOG) which was designed to determine the value of chest radiation in limited-stage small-cell lung cancer patients achieving complete response after induction chemotherapy, and to test the use of wide-field v more limited-volume radiation in patients with partial responses (PRs) and patients with stable disease (SD). The induction chemotherapy (VMV-VAC) consisted of vincristine, 2 mg intravenously (IV) every week for six doses; methotrexate, 60 mg/m2 IV days 1 and 43; VP-16, 50 mg/m2/d IV days 1 to 5 and 43 to 47; doxorubicin, 60 mg/m2 IV days 22 and 64; and cyclophosphamide, 1,000 mg/m2 IV days 22 and 64. Four hundred ninety-four patients were registered, of whom 473 were eligible. Of 466 response-evaluable patients, 153 (33%) achieved complete disease remission (CR) with chemotherapy. A total of 387 patients entered the consolidation phase of treatment after chemotherapy and response determination. CR patients were prospectively randomized to receive chest radiation, consisting of 4,800 rad administered in a split-course scheme, or to continue chemotherapy without interruption. The treatment volume was based on tumor extent before the induction chemotherapy. Maintenance chemotherapy consisted of cyclophosphamide and VP-16 administered for four cycles before a period of reinduction chemotherapy consisting of VMV-VAC as described above. Patients receiving chest radiation therapy were given the same maintenance and reinduction chemotherapy programs following completion of the chest radiation. One hundred ninety-one eligible patients achieving PR or SD status after induction chemotherapy were randomized to a preinduction treatment volume or to a postinduction reduced tumor volume, with treatment portals designed according to tumor extent before or after induction chemotherapy, respectively. After completion of the entire treatment plan, there were 218 (47%) CRs and 121 (26%) PRs. These figures represent the greatest response achieved at any point in the treatment program. The median survival for all eligible patients was 57 weeks (74 weeks for CRs). Overall survival for CR patients was not different for patients who did or did not receive chest radiation. However, patterns of tumor relapse were affected by the chest radiation, as 38 of 42 relapsing patients who did not receive radiation had intrathoracic recurrences in comparison to only 20 of 36 radiated patients.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3031229 TI - Effect of methylprednisolone on radiotherapy of F344 rats with avian sarcoma virus induced gliomas. AB - We have examined the impact of methylprednisolone acetate (MPA) on survival of F344 rats that were bearing avian sarcoma virus (ASV)-induced gliomas and that were treated optimally with radiotherapy. Toxicity of MPA (dose range of 0.2-5.0 mg/kg X 7 over 3 weeks) was first established in non-tumor bearing rats as assessed by their relative failure to gain weight. Doses of 2.0 or 5.0 mg/kg X 7 caused animals to be 21.8 or 43.9%, respectively, underweight compared with vehicle controls. In rats bearing ASV-induced gliomas, treatment with 3,000 cGY (nine fractions over a 3-week period) alone or with 0.2 or 1.0 mg MPA/kg (X 6 during the 3-week radiotherapy course) produced a significantly prolonged survival compared with that of untreated, tumor bearing rats. However, MPA did not enhance survival when given concurrently with radiotherapy; indeed, at the higher of these two doses, median survival of tumor-bearers was slightly less than with radiotherapy alone. This trend towards interference with the beneficial effects of radiotherapy was more pronounced with the highest dose of MPA studied, 5.0 mg/kg body weight X 6. These animals had a median survival time that was significantly less than that of tumor-bearers receiving radiotherapy alone, but not significantly different from untreated rats with gliomas. The possible significance of these observations is discussed. PMID- 3031230 TI - Differential effects of tetanus toxin on inhibitory and excitatory synaptic transmission in mammalian spinal cord neurons in culture: a presynaptic locus of action for tetanus toxin. AB - Tetanus toxin reduces monosynaptic inhibitory and excitatory synaptic transmission in mouse spinal cord neurons in culture. Inhibitory transmission is preferentially reduced by the toxin; however, excitatory transmission is also ultimately reduced and blocked by the concentrations of toxin used in these studies. Recordings from monosynaptically connected cell pairs revealed a marked diminution in amplitude of evoked monosynaptic inhibitory postsynaptic potentials coincident with the onset of convulsant action at a time when evoked monosynaptic EPSPs were relatively unaffected. Increased polysynaptic excitation occurred as a result of diminished inhibition. This supports the reduction of inhibition as an important mechanism in the convulsant action of tetanus toxin. Quantal analysis of the late effects of tetanus toxin on the monosynaptic excitatory postsynaptic potential revealed a reduction in quantal number with no reduction in quantal size, thus demonstrating a presynaptic locus of action for the toxin on spinal neurons. PMID- 3031231 TI - Excitatory amino acid receptors of rod- and cone-driven horizontal cells in the rabbit retina. AB - Intracellular recordings were obtained from horizontal cells in the superfused retina-eyecup preparation of the rabbit. Rod- and cone-dominated horizontal cells were studied using bath-applied excitatory amino acid analogues. Cone-dominated horizontal cell somas were depolarized by kainate (KA) or quisqualate (QQ) and their light responses were reduced or abolished. They were not affected by N methyl-DL-aspartate (NMDLA) at concentrations up to 2 mM or by 2-amino-4 phosphonobutyrate (APB), a selective agonist for the ON bipolar cell. When synaptic transmission was blocked with cobalt, horizontal cell somas were hyperpolarized. Under these conditions, KA and QQ caused large depolarizations suggesting that these agents have a direct action on horizontal cell somas. Excitatory amino acid antagonists such as cis-2,3-piperidine dicarboxylic acid (PDA) and kynurenic acid (Kyn) hyperpolarized horizontal cell somas to the level of the light-driven membrane potential. These antagonists blocked both the light driven responses and the depolarizing action of KA. The specific NMDA antagonist 2-amino-7-phosphonoheptanoate (AP-7) had no effect on the membrane potential or light-driven responses of horizontal cell somas. In contrast to a previous report, we found no evidence that low concentrations of NMDLA could hyperpolarize horizontal cells or act as a KA antagonist in the rabbit retina. Rod-dominated axon terminals were identified by waveform, threshold, and the presence of a large rod after-potential evoked by high light intensity. These cells were depolarized by KA and their light responses were attenuated. NMDLA and APB had no effect on these cells. The general antagonists, PDA and Kyn, hyperpolarized axon terminals and blocked their light-evoked responses. The specific NMDA antagonist, AP-7, had no effect on these cells. These results suggest that the synaptic receptors that mediate light input to both rod- and cone-dominated horizontal cells are kainate or quisqualate receptors. This implies that the rod and cone transmitters of the rabbit retina are similar, with the characteristics of an excitatory amino acid, such as glutamate. PMID- 3031232 TI - Junctional transmission in fast- and slow-twitch mammalian motor units. AB - The phenomena of facilitation and tetanic potentiation of end-plate potentials (EPPs) were investigated in the predominantly fast-twitch extensor digitorum longus (EDL) and the predominantly slow-twitch soleus (Sol) muscles of the rat, using extra- and intracellular recording methods. When the quantal content of the EPPs was reduced to average values of 2-3, facilitation and potentiation could be studied in both muscles without the masking effects of neuromuscular depression. Under these conditions, the facilitation and potentiation at the neuromuscular junctions of EDL were different from those at the junctions of Sol. The potentiation during sustained activation was higher in EDL than in Sol at stimulation frequencies varying between 10 and 100 Hz. The same tendency was found for the facilitation measured by paired stimuli; the facilitation in EDL was generally higher than in Sol with interpulse intervals between 300 and 10 ms. The dependence of facilitation and potentiation on the stimulation frequency was also different in EDL and Sol. As the interpulse interval decreased from 200 to 25 ms, the facilitation in Sol increased more steeply than in EDL, whereas with further decrease from 25 to 10 ms, the facilitation in Sol did not increase at all, whereas the facilitation in EDL exhibited a substantial increase. The same general pattern was observed for potentiation: the potentiation in Sol increased more steeply than in EDL between 10 and 40 Hz (100- to 25-ms intervals) and more moderately than in EDL between 40 and 100 Hz (25- to 10-ms intervals). Another paradigm, repetitive short trains, induced a progressive increase in potentiation at the neuromuscular junctions of both muscles. The cumulative effect of potentiation was more prominent in EDL than in Sol. The differences in potentiation between EDL and Sol were also evident from intracellular recordings. These recordings also revealed that the amplitude of miniature EPPs remained unaltered in both muscles before and during the stimulus trains. Comparisons of the EPPs elicited before and at the end of tetanic trains revealed that the onset time of the EPPs was delayed in Sol but not in EDL during the train. This latency shift was also observed in the focally recorded nerve terminal potentials and end plate currents in Sol, but not in EDL. The differences in facilitation and potentiation between EDL and Sol may be due to heterogeneities of the transmitter release machinery of their nerve terminals or differences in the capability of their presynaptic nerves to conduct action potentials at high frequencies.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3031228 TI - Correlation of neuropathologic findings, computerized tomographic and high resolution ultrasound scans of canine avian sarcoma virus-induced brain tumors. AB - The location, size, geometry and neuropathological findings of anaplastic astrocytomas (AA), gliosarcomas and sarcomas induced by the avian sarcoma virus (ASV) in dogs were compared with images generated using computerized tomography (CT) and real time high-resolution ultrasound (HRUS). Seven AA showed a wide range of findings on CT. Pre-contrast CT scans showed that the tumors could be hyper, hypo, or isodense. Three of seven AA had no contrast enhancement; two of these tumors were also isodense which resulted in a false-negative CT exam. Partial enhancement was seen in one tumor. This resulted in a sensitivity of detection of 72%. Real time HRUS was able to define tumor location, size and geometry of the AA missed or incompletely imaged by CT. All tumors were hyperechoic. Inhomogeneity of the echo pattern was due to hemorrhage, cyst formation, and necrosis within the tumors. Such secondary tumor characteristics were more accurately defined by HRUS compared to CT. Vasogenic edema in the brain surrounding tumors was of low density on CT and hypoechoic or indistinguishable from normal brain on US. Similar findings were seen in six gliosarcomas, two of which were not visualized by either pre- or post-contrast enhanced CT scans (sensitivity of 66%). Sarcomas differed in that they were either hyper or isodense; none were hypodense. The area of increased density matched the tumor geometry and correlated with dense cellularity and reticulin deposition. All 13 sarcomas showed contrast enhancement (100% sensitivity), but in two tumors, contrast enhanced CT underestimated the size of the tumor. Because of the large size and multiplicity of the sarcomas, HRUS imaging was not able to resolve the entire tumor volume because of limited imaging access. Intravenously injected horseradish peroxidase (HRP) crossed the tumor blood-brain barrier (BBB) only in those tumors in which contrast enhancement was seen. These studies suggest that intraoperative HRUS imaging may be useful in detecting and delineating human AA incompletely visualized by CT. PMID- 3031233 TI - Horizontal segregation of color information in the middle layers of foveal striate cortex. AB - The present study examines the chromatic organization of foveal striate cortex in the awake monkey by means of a series of microelectrode penetrations made as perpendicular as possible to the layers. The study includes 79 penetrations and 261 cells, of which 218 were tested systematically for color selectivity. Detailed analyses are conducted on a subset of 41 penetrations, which included 164 color-tested cells, an average of four cells per penetration. The penetrations were divided into two major categories on the basis of orientation selectivity testing. One group of penetrations contained at least one nonoriented cell in the first 600 microns of microelectrode trajectory (upper layers), whereas the other group of penetrations contained only oriented cells in the first 600 microns of microelectrode trajectory. The two groups were called N (nonoriented) and O (oriented), respectively. Analysis of the color properties of cells in N and O penetrations revealed that middle layer cells in N penetrations showed poor responses to white light, and color preferences for endspectral wavelengths, i.e., red or blue. Middle layer cells in O penetrations, by contrast, responded well to white light and to midspectral wavelengths. There were two subgroups of N penetrations, characterized by predominantly red (NR) or blue (NB) sensitivity in the middle layers. O penetrations could likewise be divided up into three subgroups (OG, OY, OW), characterized, respectively, by predominant sensitivity to greenish wavelengths (490-540 nm), yellowish wavelengths (550-600 nm), or white (i.e., all colors). Despite the identification of five subgroups, similarities between NR and OY, between NB and OG, and between OY and OW subgroups might be consistent with a continuum. The middle layers of N penetrations contained a unique class of cells with excitatory responses restricted to the two spectral ends, endspectral double-peak cells. A model is proposed for the color organization of layer 4 in foveal striate cortex, with red and blue zones in register with alternate cytochrome oxidase "blobs" of layers 2 and 3, white zones in register with interblob centers, and yellow and green zones in between. PMID- 3031234 TI - Modulation of pacemaker activity by IPSP and brief length perturbations in the crayfish stretch receptor. AB - We have studied the influences of brief stretches and releases and of inhibitory postsynaptic potentials (IPSPs) on pacemaker activity of the crayfish stretch receptor (RM1). Stimuli shift or reset the ongoing rhythm. Resettings were different if evaluated in interspike intervals containing perturbations, or in succeeding ones, and are referred to as early and late, respectively. Early resetting revealed that stretches and releases or IPSPs advance and delay, respectively, the next spike. With small stretches and releases or IPSPs, effects depend on the timing of the perturbation relative to the previous spike or phase, but above a characteristic mechanical perturbation amplitude the next spike fires at a fixed latency, invariant with the phase. Of particular interest was the finding that during late resetting the first successive intervals following stretches and releases or IPSPs were longer and shorter, respectively, than the period. This led, in approximately 50% of the cases, to a gradual recovery of the original pacemaker beat in the sense that spikes fire timed as if the early rhythm shift had not occurred. In conclusion, the recent firing history is essential in determining the RM1's response. The receptor's sensitivity is a complex nonlinear and periodic function of the pacemaker activity, and the response is due to interactions between pacemaker- and perturbation-induced transmembrane ionic currents. Although several alternative mechanisms may underly beat recovery, the results suggest that at least two coupled oscillators, one perturbable and the other not, provide a better explanation than a single oscillator. The physiological significance of resettings is unknown, but the early rhythm shift may synchronize RM1s in several segments when the animal's tail is moved, and conversely recovery would reduce synchrony, with obvious influences on shared postsynaptic neurons. PMID- 3031235 TI - Low extracellular magnesium induces epileptiform activity and spreading depression in rat hippocampal slices. AB - The effect of low extracellular Mg2+ concentration ([Mg2+]o) on neuronal activity was studied in rat hippocampal slices. After 20-40 min of perfusion with Mg2+ free medium, when [Mg2+]o declined to approximately 0.1-0.4 mM, spontaneous field potentials developed in the CA1 and CA3 regions, but not in the dentate gyrus. In the CA3 pyramidal cell layer, these potentials consisted of repetitive (0.3-0.5 Hz), 40- to 120-ms-long positive deflections (2-5 mV) with superimposed population spikes. In the stratum (str.) pyramidale of the CA1 region, positive negative deflections (less than 3 mV) lasting for 30-80 ms were observed, which occurred with a frequency of 0.3-0.5 Hz. In some cases, longer lasting and rapidly recurring events were also observed. In CA3 pyramidal cells, the intracellular correlates of the field potential transients were 20- to 30-mV paroxysmal depolarization shifts (PDS) with superimposed bursts of action potentials, followed by large (greater than 10 mV), 500- to 1,200-ms-long afterhyperpolarizations (AHP). In contrast, pyramidal neurons of the CA1 area did not show PDSs; instead, sequences of excitatory postsynaptic potentials (EPSPs)/inhibitory postsynaptic potentials (IPSPs) accompanied the transient field potential changes. Occasionally, spontaneous EPSPs/IPSPs, occurring with high frequencies, could also be observed in CA1 without any field potential transients. In both hippocampal regions, the epileptiform activity evolved without significant alterations in the resting membrane potential (RMP) and input resistance (RN) of the neurons, although a 2- to 5-mV reduction in action potential threshold was noted. The spontaneous activity in Mg2+-free medium was readily suppressed by raising the extracellular Ca2+ concentration ([Ca2+]o) from 1.6 to 3.6 mM. The perfusion of 10-30 microns DL-2-amino-5-phosphonovaleric acid (2-APV), an antagonist for the glutamate receptors of the N-methyl-D-aspartate (NMDA) type, also attenuated or reversibly blocked the spontaneous activity. Surgical isolation of area CA1 from CA3 ceased the occurrence of the transients in CA1 but not in CA3. The synaptic input/output curves were shifted to the left in the absence of [Mg2+]o. Threshold intensity for eliciting population spikes was 50-75% of that in normal medium. Paired-pulse facilitation was still present near threshold, but was reduced at higher stimulus intensities. Decreases in [Ca2+]o, produced by repetitive stimulation (20-Hz/5-10 s) of the Schaffer collateral/commissural pathway and monitored with ion-selective microelectrodes in the CA1 region, were enhanced in Mg2+-free medium.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3031237 TI - Pharmacology of retinotectal transmission in the goldfish: effects of nicotinic ligands, strychnine, and kynurenic acid. AB - The goal of this study was to evaluate different neurotransmitters and their receptors that might be involved in retinotectal transmission in the goldfish. Sections of tectum were isolated and maintained in vitro while pharmacological agents were administered via the tissue bath. Field potentials were elicited by electrical stimulation of the optic nerve and recorded at 50 micron depth intervals, and profiles of current source densities (CSDs) were computed from the second spatial derivatives of these potentials. The preparations were treated with low [Ca2+]/high [Mg2+] media, various cholinergic agonists and antagonists, eserine, strychnine, or kynurenic acid, via the tissue bath. Prior to treatment, depth profiles of these in vitro field potentials and CSDs closely resembled those previously reported in vivo, including 2 prominent sink-source pairs with their sinks in the superficial optic neuropil, followed by a smaller and more prolonged sink-source pair of opposite polarity. These were rapidly and reversibly eliminated by low [Ca2+]/high [Mg2+] bathing media, and substantially reduced by 0.5 or 1.0 mM kynurenic acid. By contrast, d-tubocurarine (d-TC; up to 0.16 mM) reduced peak response amplitudes by less than 40%, eliminated the third sink-source pair, and more than doubled the duration of decay of sink-source pairs 1 and 2 in a concentration-dependent manner. Strychnine had a similar action to d-TC but was slightly more potent. The time course and amplitudes of responses were not much affected by the following nicotinic agonists or antagonists (concentrations in microM): mecamylamine, 50; dihydro-beta erythroidine, 50; nicotine, 200; tetramethylammonium, 500; ACh (protected by eserine, 20), 200; alpha-bungarotoxin, 2 microM for 2.5 hr, and 0.4 microM for up to 10.5 hr; and lophotoxin, 32 microM for up to 94 min. Eserine (20 microM) and carbachol (200 microM) increased peak response amplitudes by up to 80% within 5 10 min, and amplitudes remained elevated during 20-33 min of continued treatment. The onset of the effects of d-TC, strychnine, and kynurenic acid began in 5-10 min and was completed in 30 min or less, indicating that test substances could adequately penetrate into the interior of the isolated sections of tectum. The failure of these cholinergic ligands to prevent postsynaptic responses indicates that excitatory retinotectal transmission does not depend on an intact nicotinic (or other cholinergic) system, as previously proposed. The action by kynurenic acid suggests the involvement of an excitatory amino acid neurotransmitter in retinotectal transmission.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3031236 TI - Activators of adenylate cyclase and cyclic AMP prolong calcium-dependent action potentials of mouse sensory neurons in culture by reducing a voltage-dependent potassium conductance. AB - The effects of compounds that activate adenylate cyclase and of cAMP on calcium dependent action potentials recorded from mouse dorsal root ganglion neurons were assessed. Application of compounds that stimulate the adenylate cyclase system (forskolin, cholera toxin, and prostaglandin E1) increased action potential duration with an associated decrease in afterhyperpolarization. An adenylate cyclase inhibitor, 2',5'-dideoxyadenosine, partially inhibited the responses to forskolin and cholera toxin. cAMP analogs mimicked the effect of forskolin, and the phosphodiesterase inhibitor theophylline enhanced the response to forskolin. Following intracellular injection of the potassium channel blocker cesium, the forskolin response was reduced. Forskolin did not significantly alter resting membrane potential or conductance. The action potential responses to forskolin were voltage dependent, being reduced when the membrane was held at less negative (less than -50 mV) potentials. The data suggest that activators of adenylate cyclase and cAMP prolong calcium-dependent action potentials by blocking a voltage-dependent potassium conductance that is responsible, in part, for action potential repolarization and that inactivates at membrane potentials less negative than -50 mV. PMID- 3031238 TI - Demonstration of the retrograde transport of nerve growth factor receptor in the peripheral and central nervous system. AB - NGF acts on responsive neurons by binding to specific NGF receptors on axonal termini, after which a critical biochemical signal is retrogradely transported to the cell body. The identity of the signal(s) is unknown; candidates include NGF itself or some other "second messenger." A possible second messenger is the NGF receptor. As a first step in assessing the possible role of NGF receptor in the generation of the NGF-dependent signal, and in understanding the economy of NGF receptor synthesis and utilization, we determined whether the NGF receptor is retrogradely transported. Using immunohistochemical staining with a monoclonal antibody (192-IgG) against rat NGF receptor, we looked for accumulation of NGF receptor molecules distal (retrograde transport), as well as proximal (anterograde transport), to sites of axonal ligation or transection. By 10-12 hr in both the ligated sciatic nerve and the lesioned fimbria-fornix, accumulated NGF receptor was detected proximal and distal to the ligation/lesion site. The transported receptor presumably was located in sympathetic and sensory neurons in the sciatic nerve and in forebrain cholinergic neurons projecting from the medial septum to the hippocampus. In both anatomical sites, accumulation of NGF receptor on the proximal (anterograde) side occurred in streams of fine axonal processes, whereas staining on the distal (retrograde) side occurred in varicose or granular configurations. These results raise the possibility that the NGF receptor has a role in the mechanism of NGF beyond the initial binding event at the plasma membrane of the axonal terminus. PMID- 3031239 TI - The relationship between the capillary structure and hemorrhage in gliomas. AB - A 48-year-old man was admitted with the sudden onset of symptoms of stroke caused by hemorrhage in an oligodendroglioma. Despite surgery and antiedema treatment, the patient died. Histological evaluation revealed an oligodendroglioma with calcified capillaries of the retiform type. To further investigate this phenomenon, a total of 160 gliomas were reviewed: 90 glioblastomas multiforme, 30 oligodendrogliomas, and 40 astrocytomas. Sufficient data were available for clinical evaluation in 100 cases. Of these, 5% (two oligodendrogliomas and three glioblastomas multiforme) were related to clinically significant hemorrhages. Of the remaining cases, microhemorrhages were found in 53.0% of the glioblastomas, in 56.7% of the oligodendrogliomas, and in 10.0% of the astrocytomas. In each case reviewed, the capillaries were assigned to one of three groups: axial, retiform, or glomeruloid. Statistical analysis revealed a significant association between hemorrhages and retiform capillaries in all three types of tumors, except that in oligodendrogliomas the statistical significance held true when calcification of the capillaries was also present. Glomeruloid-type capillaries were only weakly associated with hemorrhages, and no association was found for axial capillaries. A large-scale prospective study is necessary to more precisely assess the role of each of the three types of capillaries in hemorrhages of gliomas. Based on data available so far, patients with glial tumors with retiform capillaries, confirmed on biopsy, should be carefully monitored to exclude possible intratumoral hemorrhage. PMID- 3031240 TI - Catecholamine-secreting paragangliomas of the base of the skull. Report of two cases. AB - Two cases of catecholamine-secreting paragangliomas of the base of the skull are described. The patients presented with uncontrollable hypertension and, after investigation, tumors were discovered in the regions of the glomus jugulare and pterygopalatine ganglion, respectively. After cardiovascular stabilization and tumor embolization, the tumors were surgically removed, with subsequent resolution of hypertension. The incidence of these tumors is discussed. PMID- 3031243 TI - Response changing and student achievement on objective tests. AB - The purposes of this study were to evaluate the types of response changes made by undergraduate nursing majors on an objective test and to provide additional data which support the limited research on this topic. Responses were evaluated for the percent of total responses, direction of the changes made, and the effects of the changes on student achievement. Approximately 70% (n = 39) of the students changed one or more responses; the mean gain was 2.47 items. Major findings include significant differences in overall achievement on the test between response changers and nonresponse changers (t = 2.14; p less than .03); significance (X2 = 41.67; p less than .001) in the direction of the changes made by students; and significant (X2 = 19.463; p less than .0000) gain in the achievement of students who changed responses. The results of this study support the previous, but limited, research in the investigation of response changing among students. The limitation of this study and implications for future research are also presented. PMID- 3031241 TI - Intracranial metastases from malignant spinal-cord astrocytoma. Case report. AB - A patient with postoperative intracranial seeding from a malignant spinal-cord astrocytoma is presented. This case is compared with 17 previously cited cases of intracranial dissemination from spinal-cord astrocytoma. Factors associated with tumor dissemination include histological malignancy, proximity of the tumor to cerebrospinal fluid (CSF) pathways, and surgical manipulation. Hydrocephalus with infiltration of the basal cisterns also appears to be a consistent feature in these patients. Cytological studies of the CSF in this and previous cases were noted to be misleading, whereas intravenous contrast-enhanced cranial computerized tomography was invaluable for diagnosis of tumor dissemination in each case. Prophylactic irradiation of the entire neuraxis may limit intracranial metastases from malignant astrocytomas of the spinal cord. PMID- 3031242 TI - Research in nursing education: assumptions and priorities. AB - The purposes of this study were: to identify assumptions about the nature of research in nursing education, and to identify and rank order critical research questions regarding nursing education. The Delphi survey technique was used with 121 nurse educators responding to three survey rounds. The panel concurred with the assumptions that research in nursing education can and should meet criteria for scientific merit, should not be viewed as secondary in importance to nursing practice research, should emphasize the clinical nature of nursing, and needs to be less fragmented. Priorities for research included the following topics: integration of research findings into nursing curricula, development of problem solving skills, approaches to clinical teaching, and level of practice of graduates of different basic preparations. PMID- 3031244 TI - Nurses' attitudes toward the advantages of master's degree preparation in nursing. AB - The first objective of this study was to measure attitudes and opinions regarding master's degree preparation in nursing. The second objective was to identify important characteristics of graduate nursing education programs. The researcher developed Educational Attitude Questionnaire was mailed to a random sample of members of the Association of Rehabilitation Nurses and the Oncology Nursing Society. Five hundred twenty-four completed questionnaires were used for data analysis. Study findings revealed the bachelor's prepared nurses surveyed held a generally positive attitude toward graduate education in nursing. Fifty percent of the subjects indicated an intention to obtain a masters degree in nursing. For the majority of subjects the characteristics of graduate programs considered most important included faculty with high academic standards who offer challenging specialty area programs on a flexible time schedule. PMID- 3031245 TI - A comparison of role conceptions and role deprivation of baccalaureate students in nursing participating in a preceptorship or a traditional clinical program. AB - In this study, the professional and bureaucratic role conceptions and role deprivation of students participating in a preceptorship program were compared to those in a traditional faculty-supervised clinical group. The role conceptions and role deprivation of nursing faculty and preceptors were also examined. One hundred eighteen students in a two-year upper division baccalaureate program participated in this study. The Corwin Nursing Role Conception tool was administered. Results indicated no differences in role conceptions or role deprivation in students participating in the preceptorship program and those who did not. There were no differences in the developmental pattern of role conception and role deprivation during two years of nursing education. Also, nursing faculty had a significantly higher professional role conception and a significantly lower bureaucratic role conception than preceptors but there were no differences in role deprivation between the two groups. PMID- 3031247 TI - A public school-based referral system for psychiatric mental health nursing experiences. AB - Numerous advantages are anticipated with the implementation of the school-based referral system as a method of providing outpatient clinical experiences for a group of Psychiatric/Mental Health Nursing students. The school setting, when used as a clinical site, can provide valuable learning experiences where the student can apply the nursing process with individuals, families and groups. This multi-dimensional involvement reinforces family systems theory as well as group concepts. The nature of the students' experiences through the school-based referral system gives them the opportunity to increase their knowledge and understanding of the three levels of prevention: primary, secondary, and tertiary. The referrals generally constitute secondary intervention, with early recognition/intervention being the focus. Students are also encouraged to assess for factors that would place a child at risk for the development of other problems. With the identification of such factors, primary intervention strategies can be structured and implemented to reduce the child's risk. In some cases, students are involved in tertiary prevention, with a rehabilitative approach employed to assist individuals/families cope with a long-term problem situation. This expanded clinical experience facilitates the application of leadership concepts and reinforces the collaborative role of the nurse in the multidisciplinary provision of client services. Since the Psychiatric/Mental Health component of the curriculum is in the senior year, involvement in such a clinical experience also provides the students with a positive stimulus for the assumption of their professional roles and an opportunity to synthesize all previous learning. PMID- 3031248 TI - Legislated articulation of credits: initial impact of mandated career upward mobility in Arkansas nursing programs. PMID- 3031246 TI - The generic videodisc: an innovative technology in nursing education. AB - Generic videodisc is a resource that we have used to expand the clinical experience available to students in a specialized graduate curriculum. The versatility of this resource has made it equally valuable to undergraduate students, as well as students and professionals in other health-related disciplines. In contrast to usual media, the general disc has no fixed message that limits its use to one philosophy or educational objective. Educators and learners can tailor this resource to their own goals. PMID- 3031249 TI - Curriculum redesign to improve psychosocial assessment skills of first semester baccalaureate nursing students. PMID- 3031250 TI - Faculty workload measures: the time is right. PMID- 3031251 TI - Teaching the dynamics of effective oral presentations. PMID- 3031252 TI - Intestinal structural changes in African green monkeys after long term psyllium or cellulose feeding. AB - Intestinal structure of male adult African Green monkeys (Cercopithecus aethiops ssp vervets) was studied after 3 1/2 yr of consuming diets containing 10% psyllium husk or cellulose. Scanning electron microscopy (SEM) identified mild damage (cellular swelling and disarray, and microvillar denudation and disarray) at villous tips throughout the small intestine in the psyllium-fed monkeys. The cellulose group had similar duodenal damage. Differences were not found in colons by SEM. By light microscopy, jejunum had shorter villi with psyllium feeding, based upon villous height (P less than 0.05), and length around a sectioned villus (P less than 0.1), but not based upon the number of enterocytes per villus. Jejunal and ileal circular and longitudinal muscle layer thicknesses were increased in psyllium-fed monkeys. Colonic mucosal height was significantly (P less than 0.05) reduced and muscle layer thickness was mildly reduced in the psyllium-fed monkeys. Group differences were not found in intestinal weight or length or in the weight of small intestinal mucosal scrapings. Psyllium husk may cause epithelial cell loss and muscle layer hypertrophy in the jejunum and ileum and thinning of the colonic wall after prolonged feeding. PMID- 3031253 TI - Metabolizable energy in humans in two diets containing different sources of dietary fiber. Calculations and analysis. AB - The metabolizable energy (ME) of two high fiber diets, providing between 33 and 74 g dietary fiber (DF) per day, was calculated by application of Atwater's general factors (4, 9 and 4 kcal/g protein, fat and carbohydrate, respectively), Merrill and Watt's specific factors, and the British approach in which ME from carbohydrates is calculated by multiplication of the monosaccharide equivalent by 3.75 kcal/g. These factors were applied to the intakes of fat, protein and carbohydrate of 20 human subjects in two balance experiments. ME estimated by Merrill and Watt's factors agreed better with ME estimated in the balance experiment than did ME calculated by Atwater's factors or by the British approach. It was calculated that the DF in diet A, derived mainly from cereals, contributed 2.5 +/- 1.4 kcal/g to ME of the diet. The corresponding figure for DF in diet B, derived mainly from beans, vegetables and fruits, was 3.1 +/- 1.2 kcal/g. It was concluded that Merrill & Watt's factors represent the best system in current use for calculation of ME in DF-rich diets. PMID- 3031255 TI - Effects of hydrophilic fiber sources in dry rat diets. PMID- 3031254 TI - Influences of dietary restriction and age on liver enzyme activities and lipid peroxidation in mice. AB - Dietary restriction extends maximum life span in rodents by unknown mechanisms. We compared livers from 12- and 24-mo-old mice fed control (C, approximately 95 kcal/wk) or restricted (R, approximately 55 kcal/wk) amounts of diet since 3 wk of age. We hypothesized that dietary restriction might alter the activity levels of enzymes with possible relevance to aging processes. The enzymes included several xenobiotic metabolizers, radical scavengers (catalase, superoxide dismutase, glutathione peroxidase), superoxide sources (xanthine oxidase, peroxisomal beta-oxidation of palmitoyl-CoA) and glucose-6-phosphatase. Lipid peroxidation (LP) was also measured. Comparing 12- and 24-mo-old mice, the strongest diet or age effect was an increased catalase activity for group R (42% higher at 12 mo, 64% at 24 mo). LP was clearly lower in group R at 12 mo (a 30% decrease) and somewhat lower (13%) at 24 mo than in group C. Similarly, in 12-mo old C and R mice injected with either the P-450 inducer beta-naphthoflavone (beta NF in corn oil) or with corn oil alone. R mice showed higher catalase activity (40-44%) and lower LP (43-46%) in both beta-NF-injected and vehicle-injected groups. These data suggest that if free radical damage is involved in aging, it may be a particular kind of damage, that is, that in part prevented by a selective increase in catalase activity. PMID- 3031256 TI - Iodine content of various meals currently consumed by urban Japanese. AB - Various meals being currently consumed by urban Japanese were determined for iodine. The meal samples were collected in 1982 and 1984. The habitual daily home meals of 4 middle-aged Japanese living in urban areas contained 45-1,921 micrograms (mean; 362, 361, 429 and 1,023 micrograms, respectively) of iodine per day. The regular meals served in two university hospitals contained 95-287 micrograms (mean; 195 micrograms) and 89-4,746 micrograms (mean; 1,290 micrograms) of iodine per day, respectively, and the diets for diabetes mellitus contained 59-144 micrograms (mean; 96 micrograms) of iodine per day. In the daily meals containing iodine exceeding ca. 300 micrograms, some kinds of seaweeds and, in some cases, several foods containing a red food color with low iodine bioavailability, erythrosine, provided a large portion of iodine. The iodine contents of refectory meals in a university were 47-203 micrograms (mean; 113 micrograms) per meal and those of lunches in two elementary schools were 25-31 micrograms (mean; 27 micrograms) and 18-43 micrograms (mean; 36 micrograms) per lunch, respectively. These results suggest that the current daily iodine intake of urban Japanese is not great and that erythrosine elevates the iodine content of meals. PMID- 3031257 TI - Tissue response at the bone-implant interface in a hydroxylapatite augmented mandibular ridge. AB - This case demonstrates that new bone formation can occur in human HA augmented mandibular ridges. However, patient age at the time of the procedure and the length of time the implant is in place may be determining factors in the degree of osteogenesis that occurs. PMID- 3031258 TI - Histologic study of tissue response to implanted hydroxylapatite in two patients. AB - The tissue response to hydroxylapatite implants that had been used to augment deficient mandibular alveolar ridges was examined histologically in samples taken from two patients at five-months and one-year after implantation, respectively. New bone formation in the interparticular spaces was found in both cases. There was no evidence of foreign body response around the subperiosteal implanted particles. PMID- 3031259 TI - Histochemical analysis of glycoprotein and glycosaminoglycans occurring in pleomorphic adenoma of minor salivary gland. PMID- 3031260 TI - Effect of phosphorus intake in total parenteral nutrition infusates in premature neonates. AB - Phosphorus intake was evaluated in 27 appropriate weight for gestational age, critically ill neonates who required total parenteral nutrition for 2 weeks. All received approximately 30 mg/kg/d elemental calcium. The low P intake group (1.01 mmol/kg/d, 30 mg/kg/d) showed signs of phosphate depletion: hypercalciuria, hypophosphatemia, and absence of phosphaturia. The high P intake group (1.67 mmol/kg/d, 50 mg/kg/d) did not have signs of P depletion; however, they had high urinary cyclic adenosine monophosphate excretion and marked phosphaturia, suggesting secondary hyperparathyroidism. The moderate P intake group (1.34 mmol/kg/d, 40 mg/kg/d) had evidence of neither phosphate depletion nor secondary hyperparathyroidism. This phosphorus dose appears to be appropriate for the very sick, poorly growing infant receiving total parenteral nutrition. PMID- 3031261 TI - Inactive renin: a tumor marker in nephroblastoma. AB - Renin-containing cells have been identified in nephroblastoma. A prospective study of eight children with nephroblastoma has demonstrated abnormally high levels of total renin. The increase in total renin was due to increased levels of inactive renin (prorenin), rather than the active renin. These high levels decreased to normal after operation. The plasma level of inactive renin could be a useful biochemical tumor marker in nephroblastoma. PMID- 3031262 TI - Neutrophil function in localized juvenile periodontitis. Phagocytosis, superoxide production and specific granule release. AB - Patients with localized juvenile periodontitis (LJP) exhibit defective neutrophil functions to a variety of environmental and host stimuli. It is not clear, however, how many of the measurable functions are defective and whether individual patients exhibit single or multiple dysfunctions. The purpose of this study was to evaluate chemotaxis, phagocytosis, specific granule release and superoxide production in a group of 23 previously unreported LJP patients. Our results indicate that all 23 of these LJP patients exhibited chemotaxis depression to N-formyl-L-methionyl-L-leucyl-L-phenylalanine (FMLP) and endotoxin activated serum (EAS). Smaller groups from the 23 chemotactically defective LJP group were used to test other function due to inability to obtain sufficient quantities of blood. Fourteen of 14 LJP patients tested exhibited defective phagocytosis. Ten LJP patients were evaluated for specific granule release, and 14 LJP patients were evaluated for superoxide production. Both granule release and superoxide production were found to be normal in chemotactically defective LJP patients. Since both defective and normal responses noted in the same neutrophil populations are mediated by the same receptor, it is hypothesized that the cellular defect lies in a post receptor pathway. PMID- 3031263 TI - Synergistic anticellular and antiviral activities of human recombinant interferon gamma and -beta. AB - The synergism of anticellular and antiviral activities of recombinant human interferon-gamma (ReIFN-gamma) and recombinant human interferon-beta (ReIFN-beta) was examined in vitro using human melanoma SK-MEL-28 cells. Some differences were detected in the kinetics of anticellular activity between both IFNs, namely the inhibitory effect of ReIFN-beta occurred earlier than that of ReIFN-gamma. Significant synergism was detected in the anticellular activity of both IFNs when growth curves and isobolograms were examined. A difference between ReIFN-gamma and ReIFN-beta was also detected in antiviral activity. The antiviral activity of ReIFN-gamma against vesicular stomatitis virus (VSV) was significantly weaker than that of ReIFN-beta, even though both IFNs exhibited almost equivalent antiviral activities against Sindbis virus. However, ReIFN-gamma and ReIFN-beta exhibited synergistic antiviral activities against both VSV and Sindbis virus. The analysis of cell cycle distribution by flow cytometry revealed that there were some differences in the distribution pattern between cells treated with ReIFN-gamma alone, ReIFN-beta alone, or ReIFN-gamma and ReIFN-beta in combination. ReIFN-beta induced a prolongation or accumulation of S phase, whereas the effect of ReIFN-gamma was cycle-nonspecific. The combination of ReIFN gamma and ReIFN-beta induced a decrease of G1 phase and an increase of G2M phase. These results suggest that ReIFN-gamma and ReIFN-beta used in combination were more effective in inhibiting the growth of human tumor cells and the proliferation of viruses than IFN used individually. PMID- 3031264 TI - Diurnal variation of cation pump enzyme activity in pineal and seven other rat brain regions. AB - Adult female Long-Evans rats were maintained on an automatically regulated artificial lighting schedule of light:dark (L:D) 14.5:9.5 for 12 wk. After sacrifice at 0630, 1130, 1600, 1800, 2000, 2200, 0230, or 0400, the pineals were removed, weighed, and assayed for N-acetyltransferase (NAT), melatonin, Mg++ paranitrophenylphosphatase (pNPPase), and K-pNPPase activity. The brains were quickly dissected into the following areas: cerebellum, superior colliculi, inferior colliculi, visual cortex, auditory cortex, sensorimotor cortex, and the hypothalamic area around the suprachiasmatic nucleus. These regions were weighed and 10% sucrose homogenates were prepared for determinations of protein, Mg++ pNPPase, and K+-pNPPase activity. Pineal melatonin rose over six-fold from 144 +/ 70 pg/gland at 1130 to 981 +/- 173 pg/gland at 0230. Similarly, pineal NAT activity rose over 11-fold, from 119 +/- 12 pmol/gland/h to 1315 +/- 232 pmol/gland/h at the same times. K+-pNPPase activity rose by about two-thirds, from 133 +/- 12.8 nmol/gland/h to 224 +/- 22.3 nmol/gland/h from 1600 to 0230. However, when expressed per mg protein, these differences in pNPPase activity were not significant. There were no significant daily rhythms discernible in any of the seven other brain regions across these times. We conclude that cation pump enzyme activity varies only slightly with time in the rat brain and pineal gland, in spite of definite daily rhythms of pineal melatonin and NAT activity. PMID- 3031265 TI - [High-performance liquid chromatography of organosulfur compounds using the postcolumn ligand exchange reaction with iodoplatinate. Application to the simultaneous determination of (2R,4R)-2-(o-hydroxyphenyl)-3-(3-mercaptopropionyl) 4-thiazolidine carboxylic acid (SA446), an angiotensin converting enzyme inhibitor and its urinary metabolites]. PMID- 3031266 TI - [Studies on biological damage by active oxygen. I. Protective effect of various hydroxyl radical scavengers against alloxan-induced diabetes]. PMID- 3031267 TI - [Effect of eicosapentaenoic acid and arachidonic acid on mouse peritoneal exudate cells and its characteristics]. PMID- 3031268 TI - Effect of N-acetyl-DL-homocysteine thiolactone and 2,3-dimercaptosuccinic acid on the restoration of alkaline phosphatase in the nervous system of rat during methylmercury toxication. AB - The study deals with alkaline phosphatase (ALK) changes in methylmercury chloride (MMC) and the antagonists, N-acetyl-DL-homocysteine thiolactone (NAHT) and 2,3 dimercaptosuccinic acid (DSA) treated rats. A daily dose of 10 mg/kg of MMC was given to the animals for two days, seven days, and fifteen days. The animals were sacrificed on third day, eighth day, fifteen day (kept for one week without treatment) and sixteenth day. The antagonists (40 mg/kg body weight) were also applied simultaneously except in third group animals where these agents were administered from 8th-14th days. Study reveals progressive decreased activity of the enzyme in all the animal groups with increasing the duration of the dose except two days treated animals. NAHT restored the enzyme level in all the groups except in fifteen days MMC treated animals, while DSA was less effective in all the groups except in two days MMC treated animals. The significance of inhibition of the enzyme in relation to methylmercury toxication has been discussed in detail. PMID- 3031269 TI - Pyrazolo[4,5-c]quinolines. 3. Synthesis, receptor binding, and 13C NMR study. AB - Some 1-aryl-3-methylpyrazolo[4,5-c]quinolin-4-ones, were prepared and tested for their ability to displace specific [3H]flunitrazepam binding from bovine brain membranes. The 1-meta-aryl derivatives were the compounds that bound with the highest affinity within this class. Our 13C NMR study suggested a correlation between the binding affinity and the chemical shift value of a carbon atom of the tricyclic system. PMID- 3031270 TI - Synovial sarcoma presenting as tarsal tunnel syndrome. PMID- 3031271 TI - Aldosterone-reversible decrease in the density of renal peripheral benzodiazepine receptors in the rat after adrenalectomy. AB - A statistically significant decrease in the density of peripheral benzodiazepine receptors was observed in renal membranes of rats beginning 2 weeks after adrenalectomy when compared with sham-operated controls. This decrease in peripheral benzodiazepine receptor density was manifest as a decrease in the maximum binding of two ligands, [3H]Ro 5-4864 and [3H]PK 11195, without accompanying changes in their Kd for this site. Similar changes were not seen in another aldosterone-sensitive organ, the submandibular salivary gland. The decrease in peripheral benzodiazepine receptor density observed in adrenalectomized rat renal membranes was restored to control levels after 1 week of aldosterone administration using a dose (12.5 micrograms/kg/day) that had no effect on peripheral benzodiazepine receptor density in sham-operated animals. In contrast, dexamethasone administration (50 micrograms/kg/day, 1 week) had no effect on renal peripheral benzodiazepine receptor density when administered to either adrenalectomized or sham-operated rats. Further, adrenal demedullation had no effect on renal peripheral benzodiazepine receptor density or affinity. The decrease in peripheral benzodiazepine receptor density was localized to the renal cortex and the outer stripe of the medulla by gross dissection of renal slices and renal tissue section autoradiography. The specific effect of adrenalectomy on renal peripheral benzodiazepine receptor density, the lack of direct effect of aldosterone on [3H] Ro 5-4864 binding and the localization of the change in peripheral benzodiazepine receptor density to the renal cortex and outer stripe suggest that these changes may reflect an adaptation of the renal nephron (possibly the distal convoluted tubule, intermediate tubule and/or the collecting duct) to the loss of mineralocorticoid hormones. PMID- 3031272 TI - Comparison of norepinephrine- and veratrine-induced phosphoinositide hydrolysis in rat brain. AB - Stimulation of phosphoinositide hydrolysis by depolarization with veratrine was compared to that produced by stimulation of alpha-1 adrenoceptors by norepinephrine. The phosphoinositides in rat cerebral cortex were labeled with [myo-3H]inositol and the effects of the drugs on the formation of the following inositol phosphates were determined: inositol 1-phosphate (IP); inositol 1,4 bisphosphate (IP2); mixture of inositol 1,4,5-trisphosphate and inositol 1,3,4 trisphosphate (IP3). Termination of the hydrolysis by trichloroacetic acid resulted in lower basal levels and more reproducible results than termination by water lysis or a chloroform-methanol mixture (CHCl3-MeOH). The amounts of IP and IP2 formed by a maximal concentration of veratrine were about one half of that formed by a maximal concentration of norepinephrine although the amount of IP3 formed after stimulation by veratrine was only about 10% of that produced by norepinephrine. The increase in IP was linear with time (30 min) for both norepinephrine and veratrine. Stimulation of IP2 and IP3 formation by veratrine reached a maximum at 5 min whereas that produced by norepinephrine continued to increase for 30 min. Blockade of voltage-dependent calcium channels with manganese produced nearly complete antagonism of the veratrine response while only partially antagonizing the norepinephrine response. Norepinephrine-induced IP2 formation was less sensitive to manganese than was formation of IP or IP3. These data suggest that either veratrine and norepinephrine cause hydrolysis of different pools of phosphoinositide or that the hydrolysis occurs by different mechanisms. The data also suggest that IP and IP2 may be produced directly from phosphatidylinositol and phosphatidylinositol 4-phosphate rather than solely as a metabolite of IP3. PMID- 3031273 TI - Phenylquinolines PK 8165 and PK 9084 allosterically modulate [35S]t butylbicyclophosphorothionate binding to a chloride ionophore in rat brain via a novel Ro5 4864 binding site. AB - The binding of the cage convulsant, [35S]t-butylbicyclophosphorothionate ([35S]TBPS), to a picrotoxin-sensitive site in rat cerebral cortical homogenates was used to identify and characterize the site of action of the phenylquinolines PK 8165 and PK 9084, the isoquinoline PK 11195 and the atypical benzodiazepine (BZ) Ro5 4864. These agents were found to allosterically modulate the binding of 2 nM [35S]TBPS in a pharmacologically relevant fashion. Evidence is presented to suggest that these compounds share a common site of action as modulators of [35S]TBPS binding. The relative potencies of these compounds in vitro are in the submicromolar to micromolar concentration range and correlate well with the concentrations reported to elicit specific responses in behavioral and electrophysiologic studies. Modulation of [35S]TBPS binding in vitro is affected by micromolar quantities of gamma-aminobutyric acid in a (+)-bicuculline sensitive fashion and is unaffected by the central BZ receptor "antagonist" Ro15 1788. Collectively, the evidence presented suggests the existence of a novel drug binding site that is functionally coupled to a gamma-aminobutyric acid-A receptor and a [35S]TBPS-labeled chloride ionophore. Moreover, this site is distinct from the central BZ receptor recognized by clonazepam and the high-affinity peripheral BZ binding site labeled by [3H] Ro5 4864. The hypothesis is proposed that the novel "Ro5 4864 site" identified in the present study is a functionally relevant binding site that mediates some of the pharmacologic effects of Ro5 4864, PK 8165, PK 9084 and PK 11195 in the mammalian central nervous system. PMID- 3031274 TI - Binding of [3H]3-(2-carboxypiperazin-4-yl)propyl-1-phosphonic acid to rat brain membranes: a selective, high-affinity ligand for N-methyl-D-aspartate receptors. AB - 3-(2-Carboxypiperazin-4-yl)propyl-1-phosphonic acid (CPP), a rigid analog of 2 amino-7-phosphonoheptanoic acid, has been reported as a selective N-methyl-D aspartate (NMDA) antagonist. [3H]CPP bound with relatively high affinity (Kd = 201 nM) to Triton-treated rat brain crude synaptic membranes using a centrifugation assay. Binding was saturable, reversible, heat sensitive and dependent on protein concentration. Specific binding, which represented 75 to 85% of the total counts bound, was enriched in synaptosomal and microsomal fractions of rat brain, suggesting an involvement in events related to synaptic transmission. On a regional basis, binding was highest in hippocampus, followed by cortex greater than striatum greater than cerebellum = thalamus. No specific binding could be detected in pons medulla or in liver, kidney, heart, lung and adrenal tissue. [3H]CPP binding was stereoselective for the isomers of glutamate, 2-amino-5-phosphonopentanoic acid, homocysteic acid, alpha-aminoadipic acid and N methyl-aspartate. The most potent compounds tested were L-glutamate and CPP, which were equiactive in displacing [3H]CPP. The order of activity of other excitatory amino acid receptor ligands was D-2-amino-5-phosphonopentanoic acid greater than L-homocysteic acid greater than or equal to DL-2-amino-7 phosphonoheptanoic acid = D-aspartate = L-aspartate greater than L-serine-O sulfate = D-alpha-aminoadipic acid = ibotenate greater than NMDA greater than DL 2-amino-6-phosphonohexanoic acid greater than quisqualate greater than N-methyl-L aspartate. The quisqualate- and kainate-type receptor agonists DL-alpha-amino-3 hydroxy-5-methylisoxazole-4-propionate and kainic acid, respectively, had negligible activity at 100 microM.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3031275 TI - Decrease in delta and mu opioid receptor binding capacity in rat brain after chronic etorphine treatment. AB - The modulatory effect of continued activation of opiate receptors with agonist on the receptor level was examined in current studies. Rats were rendered tolerant to etorphine by s.c. implantation of osmotic minipumps containing 3 mg/ml of etorphine for up to 7 days. During this period, there were a time-dependent increase in the AD50 values of etorphine to inhibit the tail-flick response and an increase in naloxone-precipitated withdrawal signs. When these animals and others were sacrificed and the opiate receptor binding properties were examined, it could be demonstrated that there was also a time-dependent decrease in the amount of [3H]diprenorphine specifically bound, with maximal attenuation reached 3 days after implantation. There was no alteration in alpha 2 adrenergic receptor binding. This observed decrease in [3H]diprenorphine binding was not due to the presence of nonwashed etorphine in the membrane, for acute administration of the same dose of etorphine before sacrificing did not produce an attenuation of the opiate receptor binding. Further examination of opiate receptor binding in the brain regions of cortex, midbrain and striatum revealed the greatest decrease in the amount of [3H]diprenorphine bound in striatal region after chronic etorphine treatment, a 68% decrease. When the relative decrease in mu and delta opioid receptor binding was determined by carrying out [3H]-D-Ala2, DLeu5-enkephalin binding in the presence of 1 microM morphiceptin, it was observed that, 3 days after etorphine treatment, there was a decrease in mu opioid receptor binding, with minimal change in delta opioid receptor binding in both brain regions of striatum and midbrain. Only in cortex was a decrease in binding of both receptor subtypes observed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3031276 TI - Differential ontogeny of opioid, dopaminergic and serotonergic regulation of prolactin secretion. AB - The goal of this study was to determine if the opioid system which is stimulatory to prolactin (PRL) secretion develops before the serotonergic system which regulates PRL release. The opioid and serotonergic systems were chosen for comparison because evidence exists that functional serotonergic neurons are necessary for opiate-induced PRL secretion in adult rats. Haloperidol and morphine produced a dose-related stimulation of PRL release in animals of all ages. In contrast, the serotonin agonists, quipazine and m chlorophenylpiperazine, and the serotonin-releasing drug p-chloroamphetamine produced dose-related increases in PRL release in adult rats, but not in neonatal rats. The PRL response to the serotonin precursor 5-hydroxytryptophan was potentiated by fluoxetine only in animals 15 days of age or older. PRL secretion induced by these serotonergic agents was blocked by cyproheptadine, a serotonin receptor antagonist. Unlike PRL, corticosterone and growth hormone secretion were stimulated by quipazine and 5-hydroxytryptophan plus fluoxetine in both adult and neonatal rats. These findings suggest that stimulatory opioid control of PRL secretion and the dopaminergic mechanism which tonically inhibits PRL release are intact in the neonatal rat. In contrast, the stimulatory serotonergic mechanism is not functional until between 10 to 15 days of age. This late maturation appears to be specific to the serotonergic neurons regulating PRL release because the corticosterone and growth hormone responses to serotonergic stimulation develop early in ontogeny. PMID- 3031277 TI - Sensitivity of alpha-1 adrenoceptor-mediated pressor responses to inhibition by Ca++ entry blockers in the pithed rat: arguments against the role of receptor reserve. AB - In pithed rats, nifedipine (i.a., -15 min) maximally shifted the alpha-1 adrenoceptor-mediated log dose-pressor response curve to i.v. (-)-phenylephrine 5 fold to the right, doses of 1 and 3 mg/kg being equieffective. Phenoxybenzamine (3-1000 micrograms/kg i.v., -60 min) enhanced the potency of nifedipine, expressed as -log ID50, to inhibit the vasopressor response to (-)-phenylephrine. After treatment with 0.1, 0.3 or 1 mg/kg of phenoxybenzamine a dose of 1 mg/kg of nifedipine was sufficient to virtually eliminate the pressor responses. The potentiating effect of phenoxybenzamine was already present at a low dose of 3 micrograms/kg, which by itself had no influence on the log dose-pressor response curve to (-)-phenylephrine. It was also clearly limited, inasmuch as the -log ID50 values of nifedipine determined after treatment with 0.1, 0.3 and 1 mg/kg of phenoxybenzamine were identical and similar to the value reported for nifedipine against alpha-2 adrenoceptor-mediated vasoconstriction in pithed rats. Infusion of vasopressin to counteract the vasodilatory action of nifedipine did not affect its inhibitory potency compared to the effectiveness quantified under the conditions of reduced baseline diastolic pressure. The data support the conclusions that the resistance of the pressor response to (-)-phenylephrine (and other full alpha-1 adrenoceptor agonists) to inhibition by nifedipine (or other Ca++ channel blockers) is due to the dual nature of the Ca++ utilization in the vasoconstriction to these stimulants, i.e., transmembranous influx of extracellular Ca++ (sensitive to Ca++ channel blockers) and a mobilization of intracellular Ca++ (insensitive to Ca++ channel blockers).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3031278 TI - Supraspinal and spinal potency of selective opioid agonists in the mouse writhing test. AB - Three agonists with the highest degree of selectivity available for mu ([D Ala2,NMePhe4,Gly-ol]enkephalin, DAGO), delta ([ D-Pen2,D-Pen5]enkephalin, DPDPE) and kappa (U-50,488H, U50) opioid receptors were compared for their activity in inhibiting acetic acid-induced writhing in mice. Additionally, three reference agonists for mu (morphine), delta ([ D-Ala2,D-Leu5]enkephalin, DADLE) and kappa (ketocyclazocine, KC) receptors were also studied in this test. The agonists were given directly into the lateral cerebral ventricle (i.c.v.) or into the lumbar spinal subarachnoid space (intrathecal), and the potency of each compound was compared across injection sites and with data previously obtained in a thermal analgesic test (mouse hot-plate test). The rank order of potency for inhibition of writhing after i.c.v. administration was DAGO greater than DADLE greater than morphine greater than DPDPE; KC and U50 showed no significant activity by this route. After intrathecal administration, the compounds inhibited writhing with a rank potency order of DAGO greater than KC greater than morphine = DADLE greater than DPDPE greater than U50. All compounds were more potent in inhibiting writhing at spinal sites than at supraspinal sites; DPDPE and DAGO were 15 and 24 times more potent after intrathecal than after i.c.v. administration, respectively. The proposed delta agonists DPDPE and DADLE inhibited writhing at both spinal and supraspinal sites. Further, although the proposed kappa-acting compounds KC and U50 were effective at relatively low doses at spinal levels, these compounds lacked activity at supraspinal sites at doses not causing sedation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3031279 TI - Local anesthetics differentiate dihydropyridine calcium antagonist binding sites in rat brain and cardiac membranes. AB - Local anesthetics were used to probe differences in the binding of [3H]nitrendipine to dihydropyridine calcium antagonist binding sites on rat brain and cardiac membranes. Local anesthetics inhibited [3H]nitrendipine binding to brain and cardiac membranes with the rank order of potency, dibucaine = proadifen much greater than tetracaine greater than meproadifen greater than RAC-109 (S) greater than RAC-109 (R) greater than benzocaine. Lidocaine, procaine, piperocaine and bupivacaine produced either a small potentiation or inhibition of [3H]nitrendipine binding. Dibucaine inhibited [3H]nitrendipine binding to brain membranes (IC50, 4.9 +/- 0.5 microM) by increasing the Kd, whereas in cardiac membranes (IC50, 8.5 +/- 0.9 microM) it both increased the Kd and decreased the maximum binding site capacity of [3H]nitrendipine. The potency of dibucaine to inhibit [3H]nitrendipine binding was reduced in both tissues by monovalent (Li+ greater than Na+ = K+ = Rb+; EC50, 40-50 mM) and divalent (Ca++, Mg++ and Mn++; EC50, 10-50 microM) cations. These cations reduced the effect of dibucaine on the Kd of [3H]nitrendipine in brain and on the maximum binding site capacity of [3H]nitrendipine in cardiac membranes. Inhibition of [3H]nitrendipine binding by dibucaine was best described by high (2 microM) and low (50 microM) affinity sites. The apparent affinities of these sites, but not the fractional occupancies, were similar in brain and cardiac membranes. Na+ modulated the occupancies of these sites in brain, but not in cardiac membranes, whereas Ca++ inhibited occupancy of the high affinity site in both tissues. The effects of Li+ were similar to those of Ca++. These findings indicate that brain and cardiac dihydropyridine calcium antagonist binding sites are coupled to different allosteric effectors or exist in a different membrane environment. PMID- 3031280 TI - Catecholamine receptors involved in the inhibitory effects of dopamine on vagally stimulated gastric acid secretion and mucosal blood flow in rats. AB - The mechanisms of inhibitory effects of dopamine on the vagally stimulated gastric acid secretion and mucosal blood flow (MBF) were studied in anesthetized rats with a gastric fistula. Intravenous infusion of dopamine significantly reduced both gastric acid secretion and MBF. The inhibitory effect of dopamine on the vagally stimulated gastric acid secretion was not attenuated by sulpiride, metoclopramide or domperidone. Haloperidol abolished the inhibitory effect of dopamine on the acid secretion; it also abolished the inhibitory effect of norepinephrine on the vagally stimulated acid secretion. The inhibitory effect of dopamine on the acid secretion was abolished by phentolamine and yohimbine but not by propranolol or prazosin. Dopamine-induced reduction in the vagally stimulated gastric MBF was abolished by haloperidol and was partially antagonized by sulpiride or metoclopramide. In addition, the dopamine-induced reduction in the MBF was abolished by phentolamine and prazosin and was partially antagonized by yohimbine. These results indicate that the inhibitory effect of dopamine on the vagally stimulated gastric acid secretion is mediated by alpha-2 adrenoceptor mechanisms and that the inhibitory effect of dopamine on the MBF is, at least in part, mediated by alpha-1 adrenoceptor mechanisms. The authors did not obtain evidence for the existence of dopamine receptor-mediated mechanisms. PMID- 3031281 TI - Greater activation of smooth muscle alpha-2 adrenoceptors by epinephrine in distal than in proximal segments of rat tail artery. AB - The effects of selective blockade of alpha-1 and alpha-2 adrenoceptors on the vasoconstrictor responses to epinephrine (EPI) and norepinephrine (NOR) were compared in perfused/superfused proximal and distal segments of rat tail artery. The influence of neuronal uptake and of activation of beta adrenoceptors was also investigated. EPI was more potent in distal than in proximal segments. The antagonistic effect of idazoxan (100 nM) against EPI was greater in distal than in proximal segments, whereas the opposite result was obtained with prazosin (0.1 10 nM). No such difference were observed when NOR was used as agonist. Reducing the calcium ion concentration had a greater inhibitory effect on EPI in distal than in proximal segments. Cocaine (4 microM) increased responses to EPI and NOR to a greater extent in proximal than in distal segments. In the presence of cocaine, in proximal segments, antagonism of EPI by prazosin was reduced, whereas in distal segments, antagonism by idazoxan and the inhibitory effect of a reduction in calcium ion concentration were reduced. Propranolol (1 microM) increased responses to EPI and NOR to a greater extent in proximal than in distal segments. In the presence of propranolol, antagonism of EPI by both prazosin and idazoxan was reduced in proximal segments, and the inhibitory effect of a reduction in calcium ion concentration was lost. Forskolin (1 microM) inhibited responses to EPI and prevented the antagonistic effect of idazoxan, but not that of prazosin. From the results obtained it is suggested that smooth muscle alpha-2 adrenoceptors are distributed differently in the proximal and distal ends of the rat tail artery.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3031282 TI - Effects of synthetic adrenocorticotrophin on adrenal medullary responses to splanchnic nerve stimulation in conscious calves. AB - Right medullary and various cardiovascular responses to stimulation of the peripheral end of the splanchnic nerve have been investigated in the presence and absence of exogenous adrenocorticotrophin, ACTH1-24, (5 ng min-1 kg-1). The adrenal-clamp technique was employed in conscious calves, after the pituitary stalk had been cauterized and they had recovered from anaesthesia. The intravenous infusion of ACTH1-24 increased the plasma ACTH concentration by about 1100 pg ml-1 and right adrenal venous output of cortisol by about 400 ng min-1 kg body weight-1. Stimulation of the splanchnic nerve at 4 Hz for 10 min had no effect on either arterial plasma ACTH concentration or the adrenal output of cortisol. Closely similar amounts of both adrenaline and noradrenaline were released in response to nerve stimulation in the presence and absence of exogenous ACTH. In contrast, the fall in adrenal vascular resistance of about 40%, which normally occurred in response to splanchnic nerve stimulation, was completely abolished by ACTH. The adrenal produced relatively large quantities of met-enkephalin-containing peptides. During splanchnic nerve stimulation the output of these increased 2-100-fold, at which time free met5-enkephalin accounted for only 10-20% of total. During ACTH infusion the output of free met5 enkephalin was reduced at rest and during nerve stimulation, but that of total met-enkephalin-containing peptides was unaffected. These results indicate that ACTH or an adrenal steroid may alter the processing of proenkephalin in the adrenal medulla acutely but not total opiate secretion. Alternatively, the presence of ACTH could act by influencing the population of chromaffin cells activated by splanchnic nerve stimulation. PMID- 3031283 TI - Presynaptic opioid delta-receptors in the rabbit mesenteric artery. AB - Excitatory junction potentials (e.j.p.s) evoked by nerve stimulation were recorded from muscle cells of the rabbit isolated mesenteric artery. At 0.03 Hz the e.j.p. amplitudes were stable. When a train of fifteen pulses was applied at 0.25 Hz or at higher frequencies (0.5, 1 and 2 Hz), e.j.p.s showed an initial facilitation followed by depression. [Met5]enkephalin 0.1 and 1 mumol/l, [D Ala2,D-Leu5]enkephalin 0.1 and 1, but not 0.01 mumol/l, and [D-Pen2, L Pen5]enkephalin 3 mumol/l all depressed the e.j.p.s evoked by trains of fifteen pulses at 1 Hz. When more than one concentration was used ([Met5]enkephalin, [D Ala2,D-Leu5]enkephalin), the inhibition was concentration dependent. It was always greater for the first few e.j.p.s than for the later ones in a train. [Met5]enkephalin 1 mumol/l reduced the first e.j.p. at 1 Hz and the e.j.p.s evoked by 0.03 Hz to a similar extent. The inhibitory effect of [Met5]enkephalin 1 mumol/l on e.j.p.s persisted in the presence of yohimbine 0.3 mumol/l. Naloxone 1 mumol/l did not interfere with the effect of [Met5]enkephalin 1 mumol/l. Naloxone 10 mumol/l depressed some e.j.p.s and prevented the inhibition by [Met5]enkephalin 1 mumol/l. Neither ICI 154129 10 mumol/l nor ICI 174864 0.3 mumol/l had any effect of their own and both compounds antagonized the action of [Met5]enkephalin 1 mumol/l. Normorphine 10 mumol/l, fentanyl 1 mumol/l, ethylketocyclazocine 0.1 mumol/l, and dynorphin A(1-13) 1 mumol/l were all ineffective. Ethylketocyclazocine 1 mumol/l did not change the e.j.p.s either, but antagonized [Met5]enkephalin 1 mumol/l. [Met5]enkephalin 1 mumol/l failed to influence both the resting membrane potential of the muscle cells and the depolarizing effect of noradrenaline 3 and 30 mumol/l. We suggest that the axon terminals of post-ganglionic sympathetic neurones in the rabbit mesenteric artery possess opioid delta-, but not mu- or kappa-receptors. The activation of presynaptic delta-receptors inhibits the release of the neuroeffector transmitter. There is no evidence for any effect of co-released endogenous opioid peptides under our experimental conditions. PMID- 3031286 TI - Thyrotoxicosis and the hungry bone syndrome: a cause of postoperative tetany. PMID- 3031285 TI - The management of germ-cell testicular tumours. PMID- 3031284 TI - Potentiation by 4-aminopyridine of quantal acetylcholine release at the Torpedo nerve-electroplaque junction. AB - The effects of 4-aminopyridine (4-AP) on electrophysiological post-synaptic responses evoked by field stimulation or evoked focally using a loose patch-clamp technique, and on radiolabelled transmitter release were studied in the Torpedo electric organ. In this preparation, 4-AP had three major effects: it greatly potentiated the amount of acetylcholine (ACh) released by a nerve impulse, it prolonged the duration of the post-synaptic electroplaque current (e.c.) by several hundreds of milliseconds, and it increased the delay of responses triggered by a presynaptic action potential. Noise analysis performed at different times during the focally recorded giant response showed that it was made of a sustained release of ACh quanta. The maximum synchronous release of transmitter, expressed as the maximum number of quanta simultaneously delivered/micron2 of presynaptic membrane, was apparently not modified by 4-AP. A slightly different dose dependence was found for the effects of 4-AP on the potentiation of transmitter release and on the prolongation of the synaptic delay. The effects of tetraethylammonium (TEA) and other K+ channel blockers on these parameters were similar to those of 4-AP. Strong depolarizing pulses applied focally to a nerve ending were able to evoke a giant response even in the presence of 1 microM-tetrodotoxin (TTX). The prolongation of the discharge by 4 AP was therefore not caused by repetitive re-excitation of the nerve branches. Both the amplitude and the time course of the giant response were Ca2+ dependent. At a low Mg2+ concentration, the Ca2+ dependence of transmitter release was identical in the presence or absence of 4-AP. Paradoxically, in the presence of 4 AP, addition of 4 mM-Mg2+ considerably increased the Ca2+ dependence of release, whereas in the absence of 4-AP, Mg2+ blocked transmitter release by decreasing its sensitivity to Ca2+. This potentiating interaction between Mg2+ and 4-AP was not seen with TEA or guanidine. In conclusion, 4-AP potentiates ACh release in two different ways in the Torpedo electric organ: it promotes a sustained quantal release of transmitter during several hundreds of milliseconds without any significant change in the maximal synchronous release, it interacts with Mg2+ in such a manner that the sensitivity to Ca2+ of the nerve terminals is increased. PMID- 3031287 TI - Carcinoid of the biliary tree: a case report and review of the literature. PMID- 3031289 TI - HLA antigens in human parvovirus arthropathy. PMID- 3031288 TI - [Left adrenal pseudomass. Apropos of a case of splenorenal anastomosis by the vein of the left pillar of the diaphragm]. AB - A case of hepatocarcinoma on cirrhosis with portal hypertension is described. The anatomy of splenorenal anastomoses and their ultrasound and CT scan imaging characteristics are reviewed, and the interest of differential diagnosis from adrenal masses emphasized. Its importance is enhanced by the fact that the portal hypertension may be unrecognized as in the present case. Diagnostic effectiveness of ultrasound and CT scan imaging is discussed. PMID- 3031290 TI - Molecular structure of the dihydropyridazinone cardiotonic 1,3-dihydro-3,3 dimethyl-5-(1,4,5,6-tetrahydro-6-oxo-pyridazinyl)- 2H-indol-2-one, a potent inhibitor of cyclic AMP phosphodiesterase. AB - The cardiotonic 1,3-dihydro-3,3-dimethyl-5-(1,4,5,6-tetrahydro-6-oxo-3- pyridazinyl)-2H-indol-2-one (1, LY195115) is a potent, competitive inhibitor (Ki = 80 nM) of sarcoplasmic reticulum derived phosphodiesterase (SR-PDE). Moreover, the compound is a potent positive inotrope both in vitro and in vivo. To assist further cardiotonic drug-design studies, we have mapped the three-dimensional structure of 1 using X-ray crystallography. From a global viewpoint, this drug was essentially planar, but two small regions of nonplanarity were apparent. These involved the geminal methyl substituents in the indol-2-one moiety and the C5' methylene unit of the dihydropyridazinone ring. Because of our previous studies involving the bipyridine cardiotonics amrinone and milrinone, the conformational relationship between the plane of the phenyl ring and the horizontal symmetry plane defined by N2', C3', and C4' of 1 was of particular interest. The C6-C5-C3'-C4' dihedral angle was -2.7 degrees, whereas the C6-C5 C3'-N2' dihedral angle was 174.6 degrees. Therefore the two rings maintain a high degree of coplanarity. Compound 4, the congener of 1 possessing a completely unsaturated pyridazinone ring was also studied. In terms of inotropic activity, this compound, devoid of any puckering in the pyridazinone moiety, was equipotent with 1. Methyl substitution at the 4-position of the dihydropyridazinone and pyridazinone rings provided disparate results. Compound 2, the 4-methyl analogue of 1, was 2-fold more potent than 1, and the methyl substituent probably caused only minor perturbations in overall molecular topology. However 5, the 4-methyl analogue of the pyridazinone 4, was 4.4-fold less active than 4, perhaps as a result of methyl-induced molecular nonplanarity. PMID- 3031291 TI - Leukotriene receptor antagonists. 1. Synthesis and structure-activity relationships of alkoxyacetophenone derivatives. AB - A series of derivatives of 2,4-dihydroxy-3-propylacetophenone(1) were prepared and examined for their ability to block leukotriene D4 (LTD4) induced contraction of guinea pig ileum. Straight-chain carboxylic acids where the carboxyl group was separated from the acetophenone moiety by varying numbers of methylenes were evaluated, and maximum activity was obtained with the pentamethylene acid (6). Examination of ring substitution showed that the 2-propyl-3-hydroxy-4-acetyl substitution pattern was required for maximum LTD4 antagonist activity. Additional chain terminal groups were examined, and the acidic 5-tetrazolyl group separated from the acetophenone moiety by four to seven methylenes (26, 23, 27, 28) gave excellent in vitro and in vivo activities. Compound 26 (LY171883) had the best balance of in vitro and in vivo activity. It lacked bronchospastic activity at the doses administered and has been chosen for clinical evaluation. PMID- 3031292 TI - S-[(N,N'-diarylamidino)methyl] diethyldithiocarbamates and related amides as potential radioprotectants. AB - A series of symmetrically and unsymmetrically N,N'-diaryl-substituted amidinomethyl diethyldithiocarbamates and corresponding amides have been synthesized and tested for potential radioprotective activity. Antiradiation data for 10 amidines are presented. Half of the amidines showed low toxicity and radioprotective activity, based on both protective index (PI) and 30-day survival (greater than 35%) after lethal irradiation (800-rad X-rays). All (N arylcarbamoyl)methyl diethyldithiocarbamates investigated were nonprotective. PMID- 3031293 TI - Beta-adrenoceptor antagonist activity of bivalent ligands. 1. Diamide analogues of practolol. AB - Two series of bivalent ligands (P-X-P) containing the (R,S)-3-[(4-aminoaryl)oxy] 1-(isopropylamino)propan-2-ol pharmacophore and a connecting alpha,omega dicarbonylpoly(methylene) [X = -OC(CH2)nCO-] or alpha,omega-N,N' bis(carbonylmethylene) polymethylenediamine [X = -OCCH2NH(CH2)nNHCH2CO-] spanner were synthesized and evaluated for beta-adrenoceptor antagonist activity in rat heart and lung membrane preparations. The target compounds were obtained as a mixture of stereoisomers in modest yields by using a three to four step sequence beginning with N-benzylpractolol. The results from the competitive binding studies indicated that binding affinity increased by a factor of up to 160 by increasing the length of the group spanning the pharmacophore moieties. Modest increases in cardioselectivity were also obtained. The data suggest that further increases in spanner length and lipophilicity and optical resolution may improve the potential of a labeled bivalent beta 1-adrenoceptor antagonist to function as a myocardial imaging agent. PMID- 3031294 TI - Synthesis and pharmacological evaluation of gamma-aminobutyric acid analogues. New ligand for GABAB sites. AB - Baclofen (beta-p-chlorophenyl-GABA) is the only selective agonist for the bicuculline-insensitive GABAB receptor. We report the synthesis of new GABA analogues and baclofen analogues. In vitro, two compounds, 4-amino-3 benzo[b]furan-2-ylbutanoic acid (9g) and 4-amino-3-(5-methoxybenzo[b]furan-2 yl)butanoic acid (9h), showed an affinity for the GABAB receptor. The results obtained with racemic compounds of benzofuran structure, new for this series, and the surprising inactivity of compound 3a (4-amino-3-(4-hydroxyphenyl)butanoic acid) permit the proposal of an hypothesis for the structure-activity relationships with regard to GABAB receptor. PMID- 3031295 TI - Resolution of racemic carbocyclic analogues of purine nucleosides through the action of adenosine deaminase. Antiviral activity of the carbocyclic 2' deoxyguanosine enantiomers. AB - The action of adenosine deaminase on racemic carbocyclic analogues of 6 aminopurine nucleosides was investigated. When either racemic carbocyclic adenosine [(+/-)-C-Ado] or the racemic carbocyclic analogue [(+/-)-C-2,6-DAP-2' dR] of 2,6-diaminopurine 2'-deoxyribofuranoside was incubated with this enzyme, approximately half of the material was deaminated rapidly. From the resulting solution, the D isomers of the deaminated carbocyclic analogues (D-carbocyclic inosine, D-C-Ino, or D-carbocyclic 2'-deoxyguanosine, D-2'-CDG) and the L isomers of the undeaminated carbocyclic analogues were isolated. At higher concentrations of the enzyme, deamination of L-C-Ado and L-C-2,6-DAP-2'-dR proceeded slowly, thus also making the other enantiomers accessible. In tests in vitro against herpes simplex virus, types 1 and 2, D-2'-CDG was as active and potent as (+/-) 2'-CDG, whereas L-2'-CDG displayed only modest activity. In contrast to the previously reported high activity and potency of (+/-)-C-2,6-DAP-2'-dR against these two viruses, L-C-2,6-DAP-2'-dR was inactive. PMID- 3031296 TI - Synthesis of 6-substituted beta-carbolines that behave as benzodiazepine receptor antagonists or inverse agonists. AB - The synthesis of the first beta-carboline, 6-(benzylamino)-beta-carboline (1c), to be devoid of a substituent at the 3-position and that still binds to benzodiazepine receptors with potent affinity is described. Furthermore, 1c proved to be a partial inverse agonist when tested in mice. Addition of the benzylamino group at the 6-position of the beta-carboline nucleus is primarily responsible for the activity of beta-carbolines 1b and 1c. The importance of the Nb-nitrogen atom for binding affinity was also demonstrated since 3 (benzylamino)carbazole (6) exhibited little or no affinity for benzodiazepine receptors in vitro, in contrast to the activity of 1c. PMID- 3031297 TI - Direct detection of haemoglobin E with MnlI. PMID- 3031298 TI - G and R banding of 11p deletions in aniridia--Wilms' tumour. PMID- 3031299 TI - Beta thalassaemia mutations in Sardinians: implications for prenatal diagnosis. AB - In this study we have characterised by oligonucleotide hybridisation and direct restriction endonuclease analysis the beta thalassaemia mutation in 494 Sardinian beta thalassaemia heterozygotes. The most prevalent mutation, accounting for 95.4% of the cases, was the nonsense mutation at codon 39. The remainder, in decreasing order of frequency, were a frameshift at codon 6 (2.2%), beta + IVS-1, nt 110 (0.4%), and beta + IVS-2, nt 745 (0.4%). This information allows prenatal diagnosis by DNA analysis to be made in the great majority of Sardinian couples at risk for beta thalassaemia. PMID- 3031300 TI - Plasmid-mediated resistance to gentamicin in Staphylococcus aureus: the involvement of a transposon. AB - Resistance to gentamicin, tobramycin and kanamycin (GmrTmrKmr) in strains of Staphylococcus aureus isolated from clinical sources in Australia is mediated by a 4.7 kb transposable element, designated Tn4001. A 2.5 kb HindIII fragment which maps symmetrically within Tn4001, and encompasses the aminoglycoside-resistance coding region, has been shown to hybridise with fragments of identical size in HindIII digests of three different GmrTmrKmr plasmids, two of which were self transmissible, from strains of S. aureus isolated in the USA. Examination by electronmicroscopy of self-annealed molecules of the North American GmrTmrKmr plasmids revealed the presence of stem and loop structures similar to those produced by Tn4001, but with shorter inverted repeats. These results suggest that GmrTmrKmr in strains of S. aureus isolated in the USA is, or once was, transposable, and that transposable elements analogous to Tn4001 may be found in isolates of GmrTmrKmr S. aureus worldwide. PMID- 3031301 TI - Antibiotic resistances and plasmids in Staphylococcus aureus from Italian hospitals. AB - A total of 473 Staphylococcus aureus isolates from six Italian hospitals was examined for susceptibility to several antimicrobial agents and for plasmid content. Methicillin-resistant S. aureus (MRSA) were characterised by a plasmid of mol. wt (10(6)) 18-22 or 25 that carried the determinants for penicillinase production, resistance to cadmium ions and resistance to tetracycline. MRSA isolates usually harboured other smaller plasmids of mol. wt (10(6)) 2.8, 2.6 and 1.65 that encoded resistance to tetracycline, chloramphenicol and erythromycin, respectively, and cryptic plasmids of mol. wt (10(6)) c. 2 and 1 were found frequently. Methicillin-sensitive S. aureus (MSSA) that produced penicillinase often carried plasmids of mol. wt (10(6)) 11 or 13. No particular difference was found in plasmid patterns of strains from the various sources. Analysis of plasmids by EcoRI digestion showed that plasmids of similar mol. wt and phenotypic characteristics may have different restriction patterns, but often share one or more fragments in common. PMID- 3031302 TI - Bacteriophages associated with multiresistant Staphylococcus aureus in Australia. AB - Nineteen multiresistant strains of Staphylococcus aureus from Australian hospitals were examined for lysogenic bacteriophage. Thirteen strains contained prophage inducible with mitomycin C. Three of these lysed completely on induction producing a phage referred to as type 1; this phage plated on S. aureus propagating strains 6, 53 and 77, which are hosts for phages of serogroup lysogroup A III, B III and F III respectively. Type-1 phage did not plate on other propagating strains representative of the other serogroup-lysogroup combinations in the International Typing Set for S. aureus. Ten strains of S. aureus lysed incompletely when treated with mitomycin C, yielding phage type 2, that plated only on propagating strain 6. The virions of phage types 1 and 2 had isometric heads and flexible tails, and the genome consisted of c. 40 kilobases of double stranded DNA. The DNA from the two phage types was different, as shown by endonuclease digestion and by hybridisation to reference phage DNAs. The remaining six S. aureus strains contained no phage inducible with either mitomycin C or ultraviolet irradiation. However, all contained type 2 DNA, as shown by Southern blotting, present presumably in a defective prophage state. Moreover, the three strains yielding type-1 phage on induction also contained type-2 DNA. Thus, type-2 DNA was found in all 19 strains of multiresistant S. aureus from geographically diverse Australian hospitals. PMID- 3031303 TI - The effect of antibiotics on the cell morphology of Legionella pneumophila. AB - Legionella pneumophila, in Buffered Yeast Extract broth, was treated for 5 h at 37 degrees C with rosaramicin, erythromycin, cefotaxime, dibekacin, penicillin, methicillin, cefoxitin, cephalothin, ticarcillin, carbenicillin or polymyxin B at near-MIC levels and above. Electronmicroscopy demonstrated morphological changes to the bacteria in some, but not all, of the antibiotic-treated suspensions. Penicillin, at 1000 micrograms/ml (40 X MIC) but not less, produced smooth bubble like structures on cell surfaces; methicillin produced rough bubble-like structures at 100 micrograms/ml (MIC) but not at 1000 micrograms/ml. In each case, these structures resembled spheroplasts. Polymyxin B induced small-bleb formation on the bacterial cell surfaces at all concentrations tested (MIC-10 X MIC). The other eight antibiotics did not induce any morphological changes at any concentration tested. PMID- 3031304 TI - Luminol- and lucigenin-dependent chemiluminescence of neutrophils: role of degranulation. AB - The role of myeloperoxidase in luminol- and lucigenin-dependent chemiluminescence of stimulated human neutrophils has been investigated using purified myeloperoxidase and anti-(human myeloperoxidase) antiserum. This antiserum has been used as a specific enzyme inhibitor to assess myeloperoxidase-dependent neutrophil functions in single preparations of cells, thus overcoming the limitations inherent in other approaches using non-specific haem-inhibitors or myeloperoxidase-deficient neutrophils. The results show that luminol-dependent chemiluminescence is largely dependent on both oxidase activity and degranulation (of myeloperoxidase), while lucigenin monitors oxidase activity independently of the extent of degranulation. Since oxidase activation can occur in the absence of degranulation, assays utilizing luminol-dependent chemiluminescence to measure oxidant generation by stimulated neutrophils should include saturating levels of exogenous myeloperoxidase to overcome this problem. PMID- 3031306 TI - Potassium transport in the rabbit renal proximal tubule: effects of barium, ouabain, valinomycin, and other ionophores. AB - Potassium fluxes in a suspension of rabbit proximal tubules were monitored using a potassium-sensitive extracellular electrode. Ouabain (10(-4) M) and barium (5 mM) were used to selectively quantitate the potassium efflux pathway (105 +/- 5 nmol K+ X mg protein-1 X min-1) and the sodium pump-related potassium influx (108 +/- 7), respectively. These equal and opposite fluxes suggest that potassium accumulation in the cell occurs mainly through the sodium pump and that potassium efflux occurs mainly through barium-sensitive potassium channels. Thus the activity of the sodium pump (Na,K-ATPase) in the basolateral membrane of the proximal tubule is balanced by the efflux of potassium, presumably across the basolateral membrane, which has a high potassium permeability. In addition, the effect of valinomycin and other ionophores was examined on potassium fluxes and several metabolic parameters [oxygen consumption (QO2), ATP content]. The addition of valinomycin to the tubules produced a net efflux of potassium which was quantitatively equivalent to the efflux produced by the addition of ouabain. The valinomycin-induced efflux was mainly due to the activity of valinomycin as a mitochondrial uncoupler, which indirectly inhibited the sodium pump by allowing a rapid reduction of the intracellular ATP. Amphotericin, nystatin, and monensin all produced large net releases of intracellular potassium. The action of the ionophores could be localized to the plasma or mitochondrial membrane and classified into three groups, as follows: those which demonstrated full mitochondrial uncoupler activity (FCCP, valinomycin), those which had no uncoupler activity (amphotericin B, nystatin); and those which displayed partial uncoupler activity (monensin, nigericin). PMID- 3031305 TI - Downregulation of cell-to-cell communication by the viral src gene is blocked by TMB-8 and recovery of communication is blocked by vanadate. AB - The viral src gene downregulates junctional communication, closing cell-to-cell membrane channels presumably by way of the phosphoinositide signal route. We show that TMB-8 [8-N, N-(diethylamino) octyl-3,4,5-trimethoxybenzoate] counteracts this downregulation in cells transformed by temperature-sensitive mutant Rous sarcoma virus: TMB-8 (36-72 microM) raises junctional permeability when applied during activity of src protein kinase, i.e., at steady permissive temperature; and TMB-8 inhibits the fall of junctional permeability, when the activity of src protein kinase gets turned on. TMB-8 also (reversibly) inhibits the growth of the cells at permissive temperature and reverses the morphological changes associated with transformation. The morphological reversal lags several hours behind the junctional-permeability reversal. Communication recovers within a few minutes when the activity of the src protein kinase is turned off (in absence of TMB-8). Sodium orthovanadate (20 microM) prevents this recovery, but it has no major effect on junctional permeability on its own. We discuss possible modes of action of these agents on critical stages of the signal route, related to intracellular Ca2+ and protein kinase C. PMID- 3031307 TI - Cyclic AMP-dependent stimulation of Na,K-ATPase in shark rectal gland. AB - Scatchard analysis of 3H ouabain bound to isolated rectal gland cells as a function of increasing ouabain concentrations produced a concave curvilinear plot that was resolved into two specific sites with either a high (I) or low (II) affinity for ouabain. Cyclic cAMP/theophylline (+/- furosemide, 10(-4) M) increased the amount of 3H ouabain bound to the high-affinity site I. Vanadate, a phosphate congener which promotes formation of the ouabain-binding state of the enzyme, mimicked the effects of cAMP/theophylline at low concentrations of ouabain, suggesting that cAMP/theophylline increases binding to site I by enhancing the rate of turnover of resident enzyme. Enhanced 86Rb uptake seen following cAMP/theophylline administration was primarily associated with increased flux through the high-affinity ouabain site, and this stimulation was not obliterated by the co-administration of furosemide. A model was presented which suggested the presence of two noninteracting pools of enzyme or isozymes which exhibit either a high or low affinity for ouabain. Cyclic AMP both stimulated turnover via site I, and modified the kinetics of binding of 3H ouabain to site II. The (ave) Kd of 3H ouabain for site II was increased from 3.6 microM (controls) to 0.5 microM (cAMP/theophylline) and the Hill coefficient was modified from 0.45 (controls) to 1.12 (cAMP/theophylline), suggesting a transition from a negative- to a noncooperative binding state. While furosemide reversed the effects of cAMP/theophylline on site II kinetics, it did not obliterate cAMP/theophylline effects on site I. This suggests that cAMP may alter the intrinsic turnover rate of this particular pool of Na,K-ATPase in shark rectal gland. PMID- 3031308 TI - Identification of the renal Na+/H+ exchanger with N,N'-dicyclohexylcarbodiimide (DCCD) and amiloride analogues. AB - Dicyclohexylcarbodiimide (DCCD) and the 5-ethyl-isopropyl-6-bromo-derivative of amiloride (Br-EIPA) have been used as affinity and photoaffinity labels of the Na+/H+ exchanger in rat renal brush-border membranes. Intravesicular acidification by the Na+/H+ exchanger was irreversibly inhibited after incubation of vesicles for 30 min with DCCD. The substrate of the antiporter, Na+, and the competitive inhibitor, amiloride, protected from irreversible inhibition. The Na+ dependent transport systems for sulfate, dicarboxylates, and neutral, acidic, and basic amino acids were inhibited by DCCD, but not protected by amiloride. An irreversible inhibition of Na+/H+ exchange was also observed when brush-border membrane vesicles were irradiated in the presence of Br-EIPA. Na+ and Li+ protected. [14C]-DCCD was mostly incorporated into three brush-border membrane polypeptides with apparent molecular weights of 88,000, 65,000 and 51,000. Na+ did not protect but rather enhanced labeling. In contrast, amiloride effectively decreased the labeling of the 65,000 molecular weight polypeptide. In basolateral membrane vesicles one band was highly labeled by [14C]DCCD that was identified as the alpha-subunit of the Na+,K+-ATPase. [14C]-Br-EIPA was mainly incorporated into a brush-border membrane polypeptide with apparent molecular weight of 65,000. Na+ decreased the labeling of this protein. Similar to the Na+/H+ exchanger this Na+-protectable band was absent in basolateral membrane vesicles. We conclude that a membrane protein with an apparent molecular weight of 65,000 is involved in rat renal Na+/H+ exchange. PMID- 3031309 TI - How do protons cross the membrane-solution interface? Kinetic studies on bilayer membranes exposed to the protonophore S-13 (5-chloro-3-tert-butyl-2'-chloro-4' nitrosalicylanilide). AB - A simple carrier model describes adequately the transport of protons across lipid bilayer membranes by the weak acid S-13. We determined the adsorption coefficients of the anionic, A-, and neutral, HA, forms of the weak acid and the rate constants for the movement of A- and HA across the membrane by equilibrium dialysis, electrophoretic mobility, membrane potential, membrane conductance, and spectrophotometric measurements. These measurements agree with the results of voltage clamp and charge pulse kinetic experiments. We considered three mechanisms by which protons can cross the membrane-solution interface. An anion adsorbed to the interface can be protonated by a H+ ion in the aqueous phase (protolysis), a buffer molecule in the aqueous phase or water molecules (hydrolysis). We demonstrated that the first reaction cannot provide the required flux of protons: the rate at which H+ must combine with the adsorbed anions is greater than the rate at which diffusion-limited reactions occur in the bulk aqueous phase. We also ruled out the possibility that the buffer is the main source of protons: the rate at which buffers must combine with the adsorbed anions is greater than the diffusion-limited rate when we reduced the concentration of polyanionic buffer adjacent to the membrane-solution interface by using membranes with a negative surface charge. A simple analysis demonstrates that a hydrolysis reaction can account for the kinetic data. Experiments at acid pH demonstrate that the transfer of H+ from the membrane to the aqueous phase is limited by the rate at which OH- combines with adsorbed HA and that the diffusion coefficient of OH- in the water adjacent to the bilayer has a value characteristic of bulk water. Our experimental results demonstrate that protons are capable of moving rapidly across the membrane-solution interface, which argues against some mechanisms of local chemiosmosis. PMID- 3031310 TI - Identification and sequence analysis of a silent gene (ushA0) in Salmonella typhimurium. AB - The evolution of new or improved enzyme specificities in prokaryotes has been proposed to involve gene duplication, followed by silencing of one of the duplicates at the transcriptional or translational level. Such "silent gene intermediates" are distinct from "cryptic" genes, which are proposed to have a different role in evolution. We describe the identification in Salmonella typhimurium of a silent gene (ushA0) using the active (homologous) ushA gene (encoding UDP-sugar hydrolase) from Escherichia coli as a probe. The ushA0 gene has been cloned and, in the multicopy state, very weak expression can be detected; the gene product was shown to be immunologically and functionally related to the enzyme from E. coli. The sequence of the ushA0 gene was found to be highly homologous to the previously determined sequence of the ushA gene, and the respective promoter and ribosomal-binding sites are also very similar. However, a presumed strong rho-independent terminator in the ushA gene is absent from ushA0; although a weak stem-and-loop structure is present in the 3' region of ushA0, its structure is atypical of rho-independent terminators. The sequence analysis also revealed an insertion-sequence like sequence at the 3' end of ushA0 with a convergent open reading frame terminating 116 base-pairs from the ushA0 stop codon. A deletion of the 5' region of the open reading frame results in increased expression of ushA0, indicating that convergent transcription plays some role in the silencing of ushA0. S. typhimurium contains a UDP-sugar hydrolase, biochemically and genetically distinct from that in E. coli, encoded by the ushB gene. Our results indicate that ushB is not strongly sequence-related to ushA0, and its gene product is not immunologically related to the ushA gene product. ushB is hence a functional duplicate of ushA and provides a rationale for the silencing of ushA0. This situation, and the DNA sequence comparison of ushA and ushA0, strongly suggests that rather than being a cryptic gene, ushA0 has been silenced recently during the evolution of S. typhimurium. PMID- 3031311 TI - Sequence of the short unique region, short repeats, and part of the long repeats of human cytomegalovirus. AB - We have determined the DNA sequence (46 kilobases) of the short unique region, the short repeat, and part of the long repeat of human cytomegalovirus strain AD169. Analysis of the sequence has revealed at least 38 possible regions that may code for protein. Many of these open reading frames show homology to each other, and five groups of homologous reading frames are identified. Half of the predicted translation products appear to be membrane proteins, and fall into two distinct classes; those that have potential signal and anchor sequences, and those that have seven potential membrane-spanning regions and appear to be integral membrane proteins. A number of the former class contain sites for N linked glycosylation and may therefore be glycoproteins. None of the 38 open reading frames shows homology to other known herpesvirus proteins. PMID- 3031313 TI - Primary structure of poliovirus defective-interfering particle genomes and possible generation mechanisms of the particles. AB - The genomes of defective-interfering (DI) particles derived from the Sabin strain of type 1 poliovirus (PV1(Sab] were characterized by nuclease S1 mapping using complementary DNA (cDNA) copies of PV1(Sab) genome as probes. The results demonstrated variety in the size and location of the deletions, which were compatible with our previous prediction. The results further indicated that the locations of the deletions were limited within the internal genome region encoding viral capsid proteins and that the deletion sites were clustered in certain areas on the genome. Sequence analysis of a number of cloned cDNAs to the DI genomes revealed that every DI genome retained the correct reading frame for viral protein synthesis. These results strongly suggested that one or all of the viral non-structural proteins might be cis-acting at least at a certain stage in viral replication. A computer search for secondary structures with regard to the deletion sites provided a possible common structure from which, supported by sequences existing on the plus or minus RNA strand of PV1(Sab), deletion regions looped out from the remaining sequences. Replicase might, therefore, skip these transiently formed loop structures with certain frequencies, resulting in the generation of DI genomes. This model could also be considered as a model for genetic recombination in these RNA genomes. Possible "supporting sequences" were also found for every rearranged site on the RNAs of influenza virus and sindbis virus. Thus, we propose a new copy-choice model, designated the "supporting sequence-loop model", for the generation of rearrangements occurring on single stranded RNA genomes. PMID- 3031312 TI - Sequential resonance assignments as a basis for the determination of a three dimensional structure of protein E-L30 of Escherichia coli. AB - Nuclear Overhauser enhancement spectra of ribosomal protein E-L30 were searched for interresidual connectivities involving peptide bond amide protons in order to establish sequential neighbourships between amino acid residues. By comparing these data with the actual amino acid sequence of the protein, sequential resonance assignments became available for almost 90% for the amino acids in E L30. With the aid of these assignments, some 30 nuclear Overhauser connectivities could be interpreted in terms of short interproton distances involving remote sites in the polypeptide chain. It turned out that these contacts between residues generated enough constraints to permit construction of a three dimensional structure for the protein. PMID- 3031314 TI - 1H nuclear magnetic resonance study of the dynamic properties of the B and Z forms of poly[d(A-br5C).d(G-T)]. AB - Poly[d(A-br5C).d(G-T)], a synthetic polynucleotide with a 50% A-T base composition, undergoes a reversible, highly co-operative transition between the right-handed B and left-handed Z conformations. The latter is stabilized at both elevated temperature and ionic strength. The B and Z-forms of poly[d(A-br5C).d(G T)] coexist in 4.6 M-NaCl at 45 degrees C. Due to slow exchange, two sets of Tim and Gim resonances are observed and can be assigned to the B and Z conformations (the chemical shifts are, respectively, Tim = 13.4, 14.1 p.p.m. (parts/million); and Gim = 11.9, 12.4 p.p.m.). Measurements of the 1H spin-lattice (R1) and spin spin (R2) relaxation rates of the exchangeable thymine (Tim) and guanine (Gim) imino protons have been used to probe the internal dynamics of the B and Z-forms of poly[d(A-br5C).d(G-T)] and the mechanism of the B-Z transition. The proton exchange behavior in the B and Z conformations is quite different. At elevated temperature, R1 for both Tim and Gim in the B conformation is dominated by exchange with the solvent, with Tim exchanging more rapidly than Gim. This demonstrates that exchange involves the opening of single base-pairs and that neighboring A-T and G-br5C base-pairs exchange independently of each other. B form poly[d(A-br5C).d(G-T)] is unusual in that there is an acceleration of the Tim exchange rate with increasing NaCl concentration. Conversion to the Z-form by addition of 4.5 M-NaCl dramatically reduces both the Tim and Gim exchange rates (estimated to be less than 2 s-1 at 70 degrees C). Thus, the G-br5C base-pair and, in particular, the A-T base-pair are stabilized in the Z conformation. By measuring relaxation rates at 45 to 50 degrees C where the B and Z-forms are in equilibrium, we find that the B-Z interconversion rates are less than two per second. In the B conformation at 25 degrees C, the dipolar contributions to the imino proton relaxation rates are about one-third of those expected on the basis of a rigid rod model for 65 base-pair fragments, a difference we assign to large amplitude (30 degrees high frequency (less than 100 ns) out-of-plane motions of the bases. Conversion to the Z conformation has little effect on the dipolar contributions to relaxation, i.e. on the internal motions.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3031315 TI - Directional control of site-specific recombination by bacteriophage lambda. Evidence that a binding site for Int protein far from the crossover point is required for integrative but not excisive recombination. AB - Phage lambda controls its integration and excision by differential catalysis of the forward and reverse reactions. The lambda Int protein is required for both directions, but Xis for excision only. To investigate the substrate requirements for directional control, we have characterized two mutations of the phage attachment site that are defective in integrative but not excisive recombination. Both of these mutations produce the same base change in the P'3 binding site for Int protein 79 base-pairs from the center of the crossover region for site specific recombination. We infer that differential utilization of this distant binding site is crucial for directional control of recombination. PMID- 3031316 TI - Independent conformational changes caused by calmodulin and calcium in the cardiac microsomal (Ca2+, Mg2+)-ATPase during its ATP hydrolysis-ATP synthesis cycle. AB - The cardiac sarcoplasmic reticulum preparation employed in this study presented two fractions of associated calmodulin, one easily removable and the other tightly bound. The latter one was resistant to treatment with EGTA and high ionic strength. The forward or hydrolytic component of the (Ca2+, Mg2+)-ATPase cycle of calmodulin-free vesicles was stimulated when preincubated with trypsin, as opposed to the inhibition of this activity when the intact or calmodulin containing vesicles were employed in parallel assays. Interestingly, the reverse reaction of the ATPase cycle or ATP in equilibrium Pi exchange reaction found in both types of vesicles, was more sensitive to exposure to trypsin. Although a drastic inhibition of this exchange reaction was observed independent of the presence of the modulator, this reaction was transiently stimulated when the calmodulin-free vesicles were preincubated with a low trypsin to protein ratio. The differential effect of trypsin upon each reaction indicates that the equilibrium between the E1 and the E2 states of the enzyme is displaced. [125I]calmodulin was found to bind equally to the Ca X E1 approximately P and the E2 - P intermediates of the isolated enzyme formed with the ATP and Pi respectively. It is suggested the formation of an independent overall conformational state for the (Ca2+, Mg2+)-ATPase in the presence of calmodulin. PMID- 3031317 TI - The effect of treatment with coenzyme Q10 on the mitochondrial function and superoxide radical formation in cardiac muscle hypertrophied by mild aortic stenosis. AB - A 40% reduction of the diameter of the ascending aorta maintained for 60 days induced the formation of a compensate cardiac hypertrophy in rabbits without changing the value of the azide insensitive Ca2+-ATPase activity in comparison to control hearts. The cardiac mitochondria isolated from constricted animals assayed in presence of glutamate and succinate did not show a change in the R.C.I. and ADP/O values in comparison to the controls, whilst the QO2 value enhanced or decreased respectively when determined with glutamate or succinate. The intramuscular injections of CoQ10 (12 mg/kg body weight/48 h) enhanced the mitochondrial CoQ10 concentrations both in the control and in the constricted animals and further increased the QO2 value determined in both groups of animals when glutamate was used as the substrate. The production of O2.- radicals by the level of the complexes I and III of the respiratory chain, did not change in the constricted animals, nor in the animals administered with CoQ10 in comparison to the control. CoQ10 augmented the rate of oxygen consumption by the submitochondrial particles only in the constricted animals. Moreover, the treatment with the coenzyme or the constriction of the aorta, did not modify the cardiac superoxide dismutase activity, but increased the glutathione peroxidase activity only in the banded animals. In addition, in the CoQ10 treated animals there was a reduction of NADH-diaphorase activity both in the control and constricted animals, while the malondialdehyde, generated during the thiobarbituric acid test, and the cardiac content of lipofuscin were decreased. PMID- 3031318 TI - Influence of cyclic nucleotides on junctional permeability in atrial muscle. AB - The influence of dB-cAMP (5 X 10(-4) M) and 8-Br-cGMP (5 X 10(-5) M; 5 X 10(-9) M) on the longitudinal diffusion of Lucifer Yellow CH along dog trabeculae was investigated. It was found that dB-cAMP enhances the diffusion coefficient of the dye from 4 +/- 0.63 X 10(-7) cm2/s (control) to 2 +/- 0.53 X 10(-6) cm2/s. Efflux studies showed that the permeability of the surface cell membrane to Lucifer Yellow is negligible ruling out the possibility that the dye moved from cell-to cell through the extracellular space. The permeability of the nexal membrane (Pnexus = 3 X 4 cm/s) was appreciably enhanced in fibers exposed to dB-cAMP (9.1 X 10(-4) cm/s). No change in the longitudinal redistribution of Lucifer Yellow CH was found with 8-Br-cGMP. The results support the hypothesis that cAMP is a modulator of junctional permeability in the normal heart. PMID- 3031319 TI - Characterization of the macronuclear DNA of different species of Tetrahymena. AB - The macronuclear DNAs from 20 different species of Tetrahymena were characterized using Alternating Orthogonal Field (AOF) gel electrophoresis. Each species has approximately 300 different macronuclear DNA molecules that range in size from about 100-2000 kb pairs. Although the individual macronuclear DNA molecules are not well resolved on an AOF gel, most species have a unique profile of macronuclear DNA. The sequences that hybridize with histone H4 (Tetrahymena) and ubiquitin (yeast) genes were identified on the separated macronuclear DNA molecules of the different species. All species have 2 histone H4 genes located on macronuclear DNA molecules of different sizes. This is consistent with the duplication of the histone H4 gene prior to the speciation events leading to the various species of Tetrahymena. The number and sizes of the macronuclear DNA molecules that hybridize with the ubiquitin probe vary from species to species. A grouping of the different species of Tetrahymena based on this hybridization pattern parallels groupings of the species based on ribosomal RNA sequences and isoenzymes. Some intraspecific variation among different strains of Tetrahymena thermophila was detected using ubiquitin and 5S ribosomal RNA as probes. PMID- 3031320 TI - In vitro expression of two proteins from overlapping reading frames in a eukaryotic DNA sequence. AB - The in vitro expression of two distinct proteins from overlapping reading frames in a sequence of rainbow trout genomic DNA has been demonstrated. In vitro transcription of DNA sequences, cloned in a plasmid under the control of Salmonella phage 6 polymerase promoter, led to the synthesis of two distinct and functional mRNAs corresponding to the protamine mRNA and also to another overlapping mRNA, termed Y. These mRNAs were translated in an mRNA-dependent rabbit reticulocyte lysate cell free system which synthesized the corresponding protein products. Similarities between the synthesized Pro-rich protein Y and three proline-rich proteins, the human salivary Pro-rich protein, the avian sarcoma virus protein P19 and the myc oncogene product, were evident and the significance of these findings is discussed. A synthetic oligonucleotide which is complementary to a sequence corresponding to a region of the Y protein mRNA, but upstream (5') of the transcribed protamine mRNA, hybridized faintly and only to trout brain RNA. However, more sensitive primer extension studies utilizing the Y specific oligonucleotide detected several Y-related mRNAs in trout brain. PMID- 3031321 TI - Nucleotide sequence and evolution of the orangutan epsilon globin gene region and surrounding Alu repeats. AB - We have mapped and sequenced the epsilon globin gene and seven surrounding Alu repeat sequences in the orangutan beta globin gene cluster and have compared these and other orangutan sequences to orthologously related human sequences. Noncoding flanking and intron sequences, synonymous sites of alpha, gamma, and epsilon globin coding regions, and Alu sequences in human and orangutan diverge by 3.2%, 2.7%, and 3.7%, respectively. These values compare to 3.6% from DNA hybridizations and 3.4% from the psi eta globin gene region. If as suggested by fossil evidence and "molecular clock" calculations, human and orangutan lineages diverged about 10-15 MYA, the rate of noncoding DNA evolution in the two species is 1.0-1.5 X 10(-9) substitutions per site per year. We found no evidence for either the addition or deletion of Alu sequences from the beta globin gene cluster nor is there any evidence for recent concerted evolution among the Alu sequences examined. Both phylogenetic and phenetic distance analyses suggest that Alu sequences within the alpha and beta globin gene clusters arose close to the time of simian and prosimian primate divergence (about 50-60 MYA). We conclude that Alu sequences have been evolving at the rate typical of noncoding DNA for the majority of primate history. PMID- 3031322 TI - Inhibition of natural killer cell function by marijuana components. AB - The extent of modulation of host resistance mechanisms by marijuana components is not fully understood. Natural killer (NK) cells are a subpopulation of lymphoid cells and are important in host resistance mechanisms against malignant cells, virus-infected cells, and possibly pathogenic bacteria and fungi. We report that the marijuana component delta-9-tetrahydrocannabinol (THC) injected into mice results in a suppression of splenic NK activity. Furthermore, THC and the hydroxylated metabolite 11-hydroxy-delta-9-tetrahydrocannabinol (11-hydroxy-THC) suppress the NK activity of cultured murine splenocytes in a dose-dependent manner (range 1 X 10(-5) to 3.2 X 10(-5) M) without diminishing NK cell viability. The hydroxylated derivative appears to possess a more potent suppressive effect, in that it suppresses at lower concentrations than THC does and requires a shorter incubation time with the effector cells for its suppressive action. Purification of NK cells by Percoll density-gradient centrifugation suggests that both cannabinoids act directly on the natural killer cell population, resulting in suppression. Studies involving target binding analysis and calcium ionophore experiments suggest that cannabinoids do not suppress NK cell killing by the inhibition of effector/target binding or by disruption of calcium ion flux. These results suggest that two principal psychoactive cannabinoids can suppress natural killer cell function by interacting directly with the killer cells and disrupting cellular events postbinding and during the programming for lysis. Furthermore, the data suggest different modes of action for THC and the hydroxylated metabolite. PMID- 3031324 TI - Augmentation and overdenture prosthesis. Function and efficacy. PMID- 3031323 TI - Short-term oral administration of polybrominated biphenyls enhances the development of hepatic enzyme-altered foci in initiated rats. AB - FireMaster BP-6 (FM), a commercial mixture of polybrominated biphenyls (PBB), has been shown to act as a tumor promoter in hepatocarcinogenesis assays in rats. Most hepatic tumor promoters must be administered for many weeks or months. Because FM is highly persistent in animal tissues, it was hypothesized that very short-term administration of FM would result in tumor promotion. Female Sprague Dawley rats weighing 185-215 g were initiated by a two-thirds partial hepatectomy followed by 10 mg diethylnitrosamine/kg body weight (BW) 24 h later. Thirty days later, rats were gavaged with FM in corn oil, at total doses of 0, 13, or 130 mg FM/kg BW. Half the dose was given on d 30, and the remaining half was given 24 h later. At 120 d after gavage the rats were killed and necropsied. Five liver sections from each animal were histochemically stained for gamma-glutamyl transpeptidase-positive enzyme-altered foci (EAF). EAF were significantly increased over control values in initiated rats given 130 mg FM/kg. In animals given 13 mg FM/kg, EAF were increased to a lesser extent but not significantly above controls. Enhancement of these EAF in initiated rats reflects tumor promoting activity. In this study, 24-h administration of FM in initiated rats was sufficient to enhance hepatic EAF measured 120 d later in an rats was sufficient to enhance hepatic EAF measured 120 d later in an initiation-promotion protocol, and a dose of 13 mg FM/kg was apparently close to a possible no-effect threshold level for enhancement of EAF. PMID- 3031325 TI - Calciumhydroxylapatite. PMID- 3031326 TI - Plasma-sprayed titanium-I.M.Z. implant. PMID- 3031327 TI - Morphometric analysis of nasopharyngeal angiofibromas. AB - Whether recurrences of nasopharyngeal angiofibromas in some way is related to the amount and composition of the vascular components was studied by estimating the volume densities of the vascular lumen, the wall of thin-walled (lined only with endothelial cells) and thick-walled vessels (several cell-layers in the vascular wall) in seven cases of recurring and five non-recurring tumors. The volume density of the vascular compartment was less in recurring compared to non recurring tumors, 4.3 +/- 2.6% and 8.0 +/- 2.6% respectively (p less than 0.05). The volume density of the wall of thin-walled vessels was 0.7 +/- 0.2% and 1.5 +/ 0.5% for recurring and non-recurring tumors respectively (p less than 0.01). Our findings suggest that nasopharyngeal angiofibromas may be subdivided into two subgroups with different clinical behavior. PMID- 3031328 TI - AIDS, otolaryngology and a case of adenoid cystic carcinoma of the parotid arising in a patient with the AIDS-related complex. AB - The incidence of acquired immunodeficiency syndrome (AIDS) and AIDS-related complex (ARC) is increasing, and as head and neck manifestations of this entity are common it is important that otolaryngologists have an up-to-date knowledge of this condition. In this paper the epidemiological aspects of the disease are reviewed and the head and neck manifestations discussed. An interesting case of adenoid cystic carcinoma of the parotid arising in a patient with ARC is presented. PMID- 3031330 TI - Aplastic crisis or erythroid hypoplasia? PMID- 3031329 TI - Testicular cancer: prognostic implications of vascular invasion. AB - In a retrospective study the primary tumors of 33 patients with seminomas and 53 with nonseminomatous germ cell tumors were re-evaluated for vascular invasion. The significance of vascular invasion was analyzed in respect to the appearance of visceral metastases and the effect of adjuvant chemotherapy. Vascular invasion was demonstrated in 27 per cent of the patients with seminomas and 53 per cent with nonseminomatous germ cell testis tumors, while visceral metastases appeared in 9 and 32 per cent, respectively. Without adjuvant chemotherapy all 13 patients with nonseminomatous germ cell testis tumors and vascular invasion had metastases, compared to only 3 of 13 without vascular invasion (p less than 0.0005). Of 9 patients with seminoma and vascular invasion 3 had tumor progression, compared to 1 of 24 without vascular invasion (p greater than 0.05). With adjuvant chemotherapy only 1 of 15 patients (7 per cent) with nonseminomatous germ cell testis tumors and vascular invasion had metastases, compared to 100 per cent of 13 without this treatment. No significant correlation was noted between pT stage versus vascular invasion and pT stage versus tumor progression. The results demonstrate the importance of vascular invasion in the staging of and choice of treatment for early nonseminomatous germ cell testis tumors. PMID- 3031331 TI - [Bedside monitoring of neuromuscular transmission only with a nerve stimulator]. PMID- 3031332 TI - [Macroscopic study of cholangiocarcinoma]. AB - Fourty three autopsy cases of cholangiocarcinoma and 18 cases of Thorotrast related cholangiocarcinoma were macroscopically studied. They were classified into two types, according to the intrahepatic location of the tumors; middle peripheral type and hilar type. Most Thorotrast-related cases (83%) were middle peripheral type, while the non-Thorotrast-related cases were hilar type (68%). According to the gross characteristics of the tumor. They were classified into four gross types: infiltrative type (54%), nodular type (21%), periductal type (18%), and diffuse type (7%). Non-specific liver cirrhosis (mixed macro and micronodular type) was associated in 4 of non-Thorotrast related cases, and the association of biliary cirrhosis was found in 3 of the non-Thorotrast cases of the hilar type. PMID- 3031333 TI - [A case of hepatocellular carcinoma associated with multiple pulmonary tumor thrombi and rupture of its right adrenal metastasis]. AB - A 54-year-old man, who had the history of a blood transfusion 29 years ago, was admitted to our hospital because of dyspnea and abdominal fullness. Physical examination revealed jaundice and massive ascites and laboratory data suggested liver cirrhosis. The high level of AFP and a CT scan indicated the association of hepatocellular carcinoma and its metastasis to the right adrenal gland. On the 21st hospital day, he suddenly complained of severe pain in the right upper quadrant and the right flank, and fell into hemorrhagic shock. Blood transfusion was given, but he died on the 24th hospital day. Autopsy revealed liver cirrhosis, accompanied by hepatocellular carcinoma with the metastasis to the right adrenal gland and multiple pulmonary tumor thrombi. Massive hemorrhaging due to rupture of the right adrenal metastasis was seen in the retroperitoneal space. PMID- 3031334 TI - [An autopsy case with a combination of hepatocellular carcinoma and cholangiocellular carcinoma]. AB - A 68-year-old male was admitted to the Kurume University Hospital with a diagnosis of hepatocellular carcinoma (HCC). Serum CEA levels ranged from 3.7 ng/ml and 6.8 ng/ml during the course. Serum AFP levels were at normal range at admission and elevated to 13,250 ng/ml 3 months before death. He died of liver failure 17 months after admission. An autopsy disclosed diffuse infiltrative tumors distributed throughout the liver with metastasis to the bilateral lungs. Histologically, tumors of the liver consisted of elements of both HCC and cholangiolocellular carcinoma. Cholangiolocellular carcinoma is believed to occur from the canals of Hering or cholangiole, and this case may provide further information as to the histogenesis of combined HCC and cholangiocarcinoma. PMID- 3031335 TI - Autologous bone marrow transplantation in acute lymphoblastic leukemia following in vitro treatment with monoclonal antibodies and with complement: analyses for two cases of failure. AB - Two children with acute lymphoblastic leukemia (ALL) received autologous bone marrow transplantation (BMT) using remission bone marrow treated in vitro with the monoclonal antibodies, CD24 (BA-1), CD9 (BA-2) and CD10 (BA-3), and with rabbit complement. In one child with second remission ALL, hematopoietic recovery after BMT was prompt but, 81 days after BMT, isolated central nervous system (CNS) relapse occurred. Bone marrow relapse developed three months later, and she died 11 months after BMT. In patients with CNS leukemia prior to BMT, as in the present case, more intensive pretransplant CNS treatment and/or a conditioning regimen may reduce the risk of relapse. In the other patient, with primary refractory ALL in first remission, marrow reconstitution was slower. The patient developed interstitial pneumonitis with pleural effusion, and died 54 days after BMT. No infectious causes could be detected by culture or from serological studies of the pleural effusion. The rationale for applying autologous BMT to children with second remission ALL and first remission refractory ALL is discussed. PMID- 3031336 TI - [Technics used in viral genetics as an alternative means of diagnosing viral diseases]. PMID- 3031337 TI - [Sensory nerve conduction velocity (SCV) between roots and tips of fingers determined by newly designed device. An application to early detection of diabetic peripheral neuropathy]. PMID- 3031338 TI - [Magnetic resonance imaging in hemangioma of the liver, with special reference to differentiation from hepatocellular carcinoma]. PMID- 3031339 TI - [Repeated transcatheter arterial embolization therapy of hepatocellular carcinoma]. PMID- 3031340 TI - [Changes in lymphokine-activated killer cell (LAK) activity in patients with hepatocellular carcinoma after transcatheter arterial embolization]. PMID- 3031341 TI - [Restriction analysis of pepsinogen I genes in the Japanese]. PMID- 3031342 TI - [Treatment of extensive-stage small cell carcinoma of the lung in aged patients]. PMID- 3031343 TI - Studies on ABO grouping of fingernails. II. Alpha-galactosidase treatment of fingernails. PMID- 3031344 TI - [Study of parenchymal lesions in renal tuberculosis by renal scintigraphy]. PMID- 3031345 TI - [Hemodynamics of esophageal varices studied by transsplenic radionuclide portography]. PMID- 3031347 TI - Effect of cold acclimation on glucagon receptors of rat white adipocytes. AB - In order to determine the role of glucagon in cold acclimation, the changes in glucagon receptor were investigated in white adipocytes from cold-acclimated rats by establishing a glucagon radioreceptor assay method for isolated white adipocytes. The following conditions were found to be appropriate for specific glucagon receptor binding assay; cell concentration of about 1 X 10(5) cells/ml, 15 min preincubation with glucagon and 30 min-reaction at 25 degrees C in the presence of bacitracin (1 mg/ml). Cold acclimation decreased the size and increased the number of epididymal white adipocytes. Cold acclimation increased the number of glucagon receptors of white adipocytes, resulting in 140% in terms of unit cell, and 260% increase per unit surface area and 210% increase per whole tissue. However, the affinity of the binding site for glucagon was not affected. The results suggested that an enhanced metabolic response of cold-acclimated rats to glucagon could be partly explained by the increased number of glucagon receptor in white adipocytes. PMID- 3031346 TI - Augmentation of catecholamine-stimulated [3H]GDP release in adipocyte membranes from exercise-trained rats. AB - The effects of exercise training on the catecholamine-stimulated [3H]GDP release in rat adipocyte membranes prelabeled with [3H]GTP and the adenylate cyclase activity were investigated. Exercise training significantly increased the release of [3H]GDP in response to (-)isoproterenol. The adenylate cyclase activity induced by a nonhydrolyzable guanine nucleotide analogue, Gpp(NH)p, was significantly greater in exercise-trained rats. PMID- 3031348 TI - Parvocellular neurosecretory neurons: converging inputs after saphenous nerve and hypovolemic stimulations in the rat. AB - Effects of saphenous nerve stimulation and hemorrhage on discharge activity of parvocellular neurosecretory (PC) cells of the paraventricular hypothalamic nucleus (PVN) were studied in anesthetized rats. The PC cells were identified in the PVN with the criteria that units recorded in the PVN showed an antidromically conducted spike after median eminence but not after posterior pituitary stimulation of 0.45 mA intensity and were demonstrated histologically to be located in the parvocellular regions of the PVN. Saphenous nerve stimulation at 4 Hz repetition and of 3 mA intensity increased the discharge rate of 13 of the 52 PC cells tested. These excited cells exhibited a synaptically mediated excitation during post-stimulus period on peristimulus time histograms constructed during 1 Hz stimulation. Hemorrhage (4 ml/kg b.w.) activated 7 of the 10 PC cells tested which had been excited by saphenous nerve stimulation. Both saphenous nerve stimulation and hemorrhage increased immunoreactive adrenocorticotropic hormone (ACTH) in the plasma in the anesthetized rats. These data support the view that synergistic potentiation of ACTH secretion reported to occur after noxious and hypovolemic stimuli depends at least in part on the interaction between afferent neural signals originating from noxious and cardiovascular receptors on their way up to corticotropin-releasing factor (CRF)-secreting PVN neurons. PMID- 3031349 TI - Role of inhibitory and stimulative effects of prostaglandins on vasopressin stimulated osmotic water flow in the toad bladder. AB - Vasopressin-prostaglandin (PG) interaction, especially the role of the inhibitory effects of PGE2 on vasopressin action, was studied using toad urinary bladders. The PGH2, at 1 X 10(-7) M, inhibited vasopressin-stimulated water flow (Marumo, 1982); PGE2 inhibited the water flow at 10(-8) M, but PGD2, PGF2 alpha, and PGI2 did not do so even at 10(-7) M. Thus, PGE2 has a physiological effect in contrast to other PGs converted from PGH2. Indomethacin enhanced both the vasopressin- and cyclic AMP-stimulated water flow across the toad bladder. However, the half maximum activation dose for vasopressin was 2 X 10(-10) M, but for cyclic AMP, as much as 3 X 10(-8) M. The PGE2 inhibited both vasopressin- and cyclic AMP stimulated water flow. However, PGE2 inhibited vasopressin action in a dose dependent manner which was not noted as a PGE2 effect on cyclic AMP action. The W 7, which is a specific inhibitor of calmodulin, suppressed cyclic AMP-stimulated water flow in a dose-dependent manner. Thus, PGE2 may suppress vasopressin stimulated water flow at a site of cyclic AMP generation under physiological conditions. Thromboxane B2 (TXB2) enhanced vasopressin-stimulated water flow but not cyclic AMP-stimulated one. Thus PGE2 and TXB2 may be concluded as negative or positive modulators of vasopressin action in the toad bladder on the step(s) as the site of cyclic AMP generation under physiological conditions. PMID- 3031350 TI - [Combined modality treatment of lung cancer: present and future]. PMID- 3031351 TI - [A case of tracheal adenoid cystic carcinoma managed by resection of 8 tracheal rings and the lateral half of the right main bronchus and primary anastomosis]. PMID- 3031352 TI - Comparison of contractile effects of sodium vanadate and ouabain in vascular smooth muscles of guinea-pigs and rats. AB - Ouabain and vanadate are known as potent inhibitors of Na, K-ATPase in various tissues including smooth muscles. Both agents showed contractile action on various smooth muscles in a similar fashion: stronger contractile action on the aortae of rats (WKY and stroke prone spontaneously hypertensive rats, SHRSP) and guinea-pigs, and weaker contractile actions on basilar and mesenteric arteries of the same animals. Time to peak tension, however, was far longer in ouabain induced contraction. Phentolamine depressed ouabain-induced contractions, while vanadate-induced contractions were not affected. Elevation of K+ concentration to 20 or 30 mM potentiated vanadate-induced contraction markely, while it potentiated ouabain-induced contraction only slightly. DIDS blocked vanadate induced contraction but showed no effect on ouabain-induced contraction. Removal of Ca abolished ouabain-induced contractions, while vanadate-induced contractions of reduced height could be observed in the absence of Ca. Verapamil depressed both ouabain- and vanadate-induced contractions of WKY and SHRSP aorte aut exhibited no effect on the guinea-pig aorta. Thus, although similarities of the action of ouabain and sodium vanadate were observed, the modes of the actions were revealed to be different in the two agents. Inhibition of Na, K-ATPase might be involved in the case of ouabain-induced contractions, and inhibition of Ca ATPase of membranous systems might be involved in vanadate-induced contraction. PMID- 3031354 TI - [Evaluation of the function of the left ventricle of the heart according to the results of radionuclide ventriculography in the diagnosis of ischemic heart disease]. AB - Radionuclide (99mTc) ventriculography demonstrated a tendency to reduced ejection fraction, mean normalized systolic expulsion rate and myocardial circular fibre shortening rate of coronary patients that was proportionate to the severity of their condition. Myocardial lesion was shown to be heterogeneous and typically involve asynergic areas. As the disease progresses, the disturbance of myocardial contractility grows more severe, and the number of hypokinetic areas increases while that of normo- and hyperkinetic areas declines. PMID- 3031353 TI - [The HLA system and Coxsackie B viral myocarditis in adults]. AB - The distribution of HLA antigens A, B and C was studied in 152 normal donors, 53 patients with Coxsackie B virus myocarditis, 35 patients with myocarditis of unknown origin, 16 coronary patients and 14 rheumatic patients with high titres of anti-Coxsackie virus antibodies. Coxsackie virus myocarditis was associated with increased occurrence of HLA antigens A3, B40, Cw2 and A28, the increase being significant (with an adjustment to the number of the tested antigens) for A3 in severe and medium myocarditis, and B40 in patients who had developed myocarditis before age 30. Antigen Cw2 was more common in myocarditis patients with high titres of anti-Coxsackie virus B4 antibodies. The distribution of HLA antigens in coronary patients was similar to that of the controls. Rheumatic patients showed increased incidence of antigen Cw2. It is suggested that carriers of histocompatibility antigen A3 may be predisposed genetically to severe or medium Coxsackie B virus myocarditis. PMID- 3031355 TI - [Mechanisms of the hypotensive action of captopril in essential hypertension]. AB - Captopril effects were studied in 27 patients with second-stage essential hypertension following administration of a single 25 mg oral dose of the drug. Blood pressure (BP), blood angiotensin-1-converting enzyme (ACE) activity, active (APR) and inactive (IPR) plasma renin levels were measured every 30 minutes for 3 hours. Before and near the end of the acute test, urinary kallikrein and catecholamine excretions were measured, and systemic and regional hemodynamic changes assessed. BP decreased in 21 patients: 11 of those had low basal APR (group 1), and 10 had normal or moderately elevated APR (group 2). The greatest hypotensive effect was observed within 1.5 hours after the administration, coinciding with the most marked ACE inhibition. There was no significant intergroup difference with respect to the extent of the hypotensive effect. No correlation was found between the hypotensive effect and the degree of ACE inhibition. All patients showed significantly decreased arteriolar tone, increased venous distensibility, decreased total peripheral resistance, and expanded end diastolic volume, while their cardiac output and heart rate remained unaffected. Hypertensive patients with low APR gave no evidence of renin angiotensin inhibition or kallikrein-kinin activation that might have accounted for the hemodynamic effect of captopril. PMID- 3031356 TI - [Role of erythrocyte membrane cholesterol in disorders of their structuro functional status in patients with ischemic heart disease]. AB - Increased molar cholesterol/phospholipid ratios, significantly increased membrane microviscosity, reduced filterability through a 3 micron-pore filter, increased aggregation and longer disaggregation time, and occasional abnormally large cells were found in erythrocyte assays of 80 coronary patients, as compared to normal subjects. It is suggested that erythrocyte microrheologic disorders are related to excessive cellular cholesterol content in coronary patients. PMID- 3031357 TI - Aminoglycoside-induced alterations of phosphoinositide metabolism. AB - There exists a strong interaction between aminoglycosides and phosphoinositides, and these membrane lipids are even considered as the drug receptors. To shed some light on the role of such an interaction in the drug nephrotoxicity, we have investigated the influence of aminoglycosides on phosphoinositide metabolism in kidney proximal tubules where these compounds accumulate. Experiments were carried out by measuring 32P labelling of phosphatidylinositol 4-phosphate (PI-P) and of phosphatidylinositol 4,5-bisphosphate (PI-P2) after incubation of homogenates of isolated proximal tubules with [gamma-32P] ATP. The treatment of rabbits with neomycin, gentamicin and amikacin (50, 50 and 300 mg/kg/day, respectively for seven days) promoted a decrease in 32P-PI-P2 and an increase in 32P-PI-P, when compared to the respective values observed in tubules from untreated rabbits. Under these conditions, the extent of modifications in lipid labelling was similar with the three drugs tested. In in vitro experiments, the exogenous addition of the above aminoglycosides to the incubation medium containing tubule homogenates from untreated rabbits also produced, in a dose dependent manner, a decrease in 32P-PI-P2 and an increase in 32P-PI-P. In the in vitro experiments, however, amikacin and gentamicin appeared to be less potent than neomycin. The results indicated moreover that phosphoinositide metabolism was more sensitive to the in vivo (vs. in vitro) action of the drugs. Phosphoinositides are involved in Ca2+ transport and/or mobilization processes, and aminoglycosides are known to interfere with the Ca2+ binding to membranes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3031358 TI - Receptors to vasopressin and other hormones in the mammalian kidney. PMID- 3031359 TI - [Changes in rickets prevention and changes in vitamin D preparations (the "new" Dekristol)]. PMID- 3031361 TI - [Stevens-Johnson syndrome as a complication of bronchography]. PMID- 3031360 TI - A quantitative receptor assay for "digitalis-like" compounds in serum. Demonstration of raised concentrations in essential hypertension and correlation with arterial blood pressure. AB - The influence of serum from patients with essential hypertension on [3H]ouabain binding to isolated (Na+ +K+)-ATPase and on the reactivity with digoxin-specific antibodies was investigated. [3H]Ouabain binding to (Na+ +K+)-ATPase was significantly decreased (P less than 0.001) by sera of 18 hypertensive patients (34.9 +/- 1.5 pmol/U) as compared with 22 normotensive controls (43.8 +/- 1.2 pmol/U). The factor, whose concentration is increased in the serum of patients with essential hypertension, competed with [3H]ouabain at isolated (Na+ +K+) ATPase. Therefore, it was possible to quantify this "digitalis-like" factor with a receptor assay in ouabain equivalents. Three times higher mean serum levels were found in hypertensive patients (234.8 +/- 48.7 nM) than in normotensive controls (76.3 +/- 9.3 nM). Deproteinization of the sera by ultrafiltration through steroid-adsorbing membranes and by boiling of acidified sera for 10 min led to a significant reduction of the inhibitory activity and to the complete loss of a difference between the sera of normotensives and hypertensives. After deproteinization by boiling for 15 min, sera of normotensives showed levels of "digitalis-like" compounds of 16.53 +/- 2.15 nM and hypertensives of 41.65 +/- 8.41 nM (P less than 0.05). Though significantly elevated concentrations of "digitalis-like" factor were measured with the receptor assay, no significant increase of digoxin-like activity could be detected with digoxin-specific antibodies in untreated serum. PMID- 3031362 TI - [Role of cyclic nucleotides in the pathogenesis of arterial hypertension]. PMID- 3031363 TI - [Changes in arterial pressure, serum lipids and thromboxane B2 after using a diet with various levels of eicosapentaenoic acid in patients with hypertension]. PMID- 3031364 TI - Glycerol-3-phosphate excretion in fructose-1,6-diphosphatase deficiency. AB - A patient aged 23 months with fructose-1,6-diphosphatase deficiency is reported. This infant demonstrated an increased urine excretion of glycerol-3-phosphate during episodes of hypoglycaemia. The excretion of this compound has not previously been described in this disease or in those disorders associated with a deficiency in one of the other three gluconeogenic enzymes associated with hypoglycaemia. Its presence in the urine from patients may be useful in diagnosis. PMID- 3031365 TI - The angiotensin I-converting enzyme. PMID- 3031366 TI - Cellular and subcellular immunohistochemical localization of angiotensin converting enzyme in the rat adrenal gland. AB - Angiotensin-converting enzyme (ACE) was localized by immunocytochemical methods in the rat adrenal gland. The three-layer peroxidase-antiperoxidase method was used with light and electron microscopy. At the cortical level, ACE immunoreactivity occurred only on the luminal side of vascular endothelial cell membranes in the adrenal capsule, whereas no ACE activity was found in the zona glomerulosa, fasciculata, or reticularis. ACE immunoreactivity was found mainly but not exclusively on the luminal side of all the types of vessels in the adrenal medulla such as capillaries, venous sinuses, and arteriae medullae. In addition, ACE was localized on the plasma membrane of chromaffin cells but never in Schwann cells nor in nerve fibers. These results provide evidence for a local production of angiotensin II at the vascular level in the adrenal gland. In addition, the presence of ACE on the plasma membrane of chromaffin cells suggests that angiotensin II or other peptides, such as bradykinin, could be metabolized here. PMID- 3031367 TI - Ultrastructural changes associated with retroviral replication in central nervous system capillary endothelial cells. AB - Widespread ultrastructural alterations in the vascular basal lamina were found in the central nervous system of symptomatic mice infected with three different neuropathogenic murine leukemia viruses. These changes appeared to progress in frequency and severity in parallel with the progression of spongiform lesions and clinical symptoms. Accumulations of pleomorphic, apparently degenerating, virions were frequently seen within areas that showed marked aberrations from the normal basal lamina ultrastructure. We suggest that the observed retrovirus-associated ultrastructural changes in the endothelial basal lamina may alter normal physiologic functions and thus play a primary role in the pathogenesis of retrovirus-induced spongiform disease. PMID- 3031370 TI - Bartholin gland carcinoma: a case report and review of the literature. PMID- 3031368 TI - Cyclic AMP inhibits increased collagen production by cyclically stretched smooth muscle cells. AB - Rabbit arterial smooth muscle cells, grown on elastin membranes which were cyclically elongated and relaxed, responded by increasing their rates of synthesis of protein and, in particular, of collagen, compared to stationary controls. Raising intracellular cyclic AMP (cAMP) levels by adding theophylline or dibutyryl cAMP to the culture medium prevented the synthetic response to cyclic stretching, but did not alter the rates of protein or collagen synthesis by stationary controls. Both synthesis and degradation of collagen by cyclically stretched cells increased in parallel such that the proportion of synthesized collagen that was degraded was similar to that found in the stationary cultures. Collagen degradation was not affected by theophylline administration to stationary cell cultures but the drug increased degradation of collagen by cyclically stretched cells. We conclude that the net production of protein, and in particular of a structural protein, collagen, by arterial smooth muscle cells subjected to the mechanical force of stretching was inhibited when intracellular levels of cAMP were raised. The results suggest that cAMP may play a role in the modulation of structural protein content of artery walls in response to changes in tensile stress. PMID- 3031369 TI - The pathogenesis of spinal cord involvement in the encephalomyelitis of mice caused by neuroadapted Sindbis virus infection. AB - Weanling mice develop an acute encephalomyelitis with high mortality after intracerebral inoculation of neuroadapted Sindbis virus. The mice develop kyphoscoliosis and hindlimb paralysis. Immunohistochemical and in situ hybridization studies have demonstrated virus in the gray matter of the brain and spinal cord. Ventral horn cells are prominently infected, providing an anatomical basis for the clinical poliomyelitis. A novel route of spread of inoculated virus within the central nervous system has been found. The virus enters the ventricular system, and then travels caudally in the central canal of the spinal cord where ependymal cells are infected. The virus subsequently spreads into the gray matter. The distribution of virus in the spinal cord is likely dependent both on variations in the susceptibility of neural cells and on this route of entry and subsequent spread. PMID- 3031371 TI - Functional identification of the perioral neuromuscular system: a signal flow diagram. AB - The components of a neuromuscular system are identified, and anatomical and physiological features of the elements are organized into a signal flow diagram. The elements of the perioral neuromuscular system are described in the context of the system flow diagram developed for a generalized neuromuscular system. Important features of commonality and contrast between limb and perioral systems, and areas of needed research are emphasized. Features of contrast include geometry and insertions of muscles, relative influence of inertial loads, and organization of inputs to motoneurons. PMID- 3031372 TI - Vasoactive intestinal peptide (VIP) stimulates aldosterone secretion by rat adrenal glands in vivo. AB - VIP acutely enhanced the plasma concentration of aldosterone (but not that of corticosterone) both in normal rats, and in rats chronically treated with dexamethasone and ACTH or captopril and angiotensin II. VIP increased aldosterone blood concentration in chronically captopril-treated animals, but not in rats in which ACTH secretion was inhibited by dexamethasone. These findings suggest that VIP is specifically involved in the stimulation of the secretory activity of rat zona glomerulosa, and that this action of VIP requires a normal level of circulating ACTH. PMID- 3031373 TI - Identification of a mineralocorticoid receptor binding substance in the urine of patients with congenital adrenal hyperplasia. AB - Increased amounts of circulating mineralocorticoid receptor binding substances presumed to be natural antagonists were previously demonstrated in congenital adrenal hyperplasia. In this study the feasibility of using urinary extracts for the identification of such binding substances was investigated. Urinary extracts from patients with the 21-hydroxylase defect did contain greater than normal amounts of mineralocorticoid receptor binding material. When subjected to chromatographic separation using a radioreceptor assay to follow the course of fractionation, a major aldosterone binding competitor was identified. On the basis of its chromatographic mobility in comparison with the labeled steroid, radioimmunoassay, ultraviolet absorption and radio-receptor assay of the native and acetylated derivative, the component was identified as 11-deoxycorticosterone and its structure confirmed by mass spectrometry. Although the major mineralocorticoid receptor binding component proved not to be an antagonist but an agonist, the results are in keeping with other evidence for overproduction of 11-deoxycorticosterone in the simple virilizing form of the disorder. Our finding did not disprove the existence of a circulating mineralocorticoid antagonist in congenital adrenal hyperplasia, but demonstrate that the major receptor binding substance in urinary extracts in that disorder is the mineralocorticoid agonist, 11-deoxycorticosterone. PMID- 3031374 TI - Influence of an estrone-desulfating intestinal flora on the enterohepatic circulation of estrone-sulfate in rats. AB - The fecal and urinary excretion of orally administered [4-14C]estrone-3-sulfate was studied in germfree (GF) rats, conventional (CV) rats and gnotobiotic rats selectively associated with estrone-desulfating and/or cecal-volume reducing microorganisms. The time required to excrete 50% of the total label recovered (t 1/2) was 22 h in CV rats vs 32 h in GF rats. Gnotobiotic rats selectively associated with a cecal volume-reducing flora (CRF rats) excreted the label even faster (t 1/2 = 13 h) than CV rats. Association of GF rats as well as CRF rats with estrone-desulfating microorganisms (termed S1 + S2 + R9 rats and CRF + S1 + S2 + R9 rats, respectively) led to a slower excretion of labeled products (t 1/2 = 38 h in S1 + S2 + R9 rats and t 1/2 = 27 h in CFR + S1 + S2 + R9 rats). Intestinal microbial desulfation also increased the relative part of the urinary excretion from 4% in GF rats to 8% in S1 + S2 + R9 rats and from 3% in CRF rats to 9% in CFR + S1 + S2 + R9 rats. We conclude that intestinal microbial desulfation enhances the enterohepatic circulation of orally administered estrone 3-sulfate. PMID- 3031375 TI - Chronic treatment with corticotropin increases the capacity of rabbit adrenocortical cells to convert pregnenolone into androgens. AB - The present study was conducted to evaluate whether the previously demonstrated enhancement in adrenocortical androgen secretion in rabbits chronically treated with ACTH results, in addition to an increased pregnenolone production, from a more efficient conversion of this precursor of steroidogenesis into androgens. To this end, the adrenocortical cells from 14 control and 14 ACTH-treated rabbits (ACTH 1-24,200 micrograms s.c. daily for 12 days) were incubated either in the presence of different concentration of ACTH or with pregnenolone added in amounts from 0.5 to 250 micrograms. The total steroidogenic potency (maximal response to ACTH) was significantly enhanced for cells from ACTH-treated animals, as was the ACTH-induced production of dehydroepiandrosterone (DHEA), DHEA-sulfate, androstenedione and testosterone. In addition the production of these androgens from given amounts of exogenous pregnenolone was also significantly increased. The maximal capacity of adrenocortical cells to convert pregnenolone into androgens averaged (for ACTH-treated vs control group) 130 +/- 34 vs 43 +/- 10 pmol for DHEA, 138 +/- 43 vs 46 +/- 14 pmol for DHEA-sulfate, 99 +/- 31 vs 10 +/- 2 pmol for androstenedione and 8.0 +/- 2.6 vs 2.4 +/- 0.3 pmol for testosterone (P less than 0.001 for all androgens). The addition of ACTH to adrenocortical cells incubated with pregnenolone did not modify the maximal capacity of conversion of pregnenolone into androgens, which was in both experimental groups similar to that documented in the absence of ACTH. Thus, while an acute stimulatory effect of ACTH on adrenocortical steroidogenesis is devoid of any influence on the activity of the post-pregnenolone pathway of androgen synthesis, the chronic exposure of adrenocortical cells to ACTH lead to increased activity of steroidogenic pathway involved in the conversion of pregnenolone into androgens. PMID- 3031376 TI - Infected mullerian adenosarcoma of the endometrium. AB - A 52-year-old postmenopausal woman presented with signs and symptoms of acute pelvic inflammatory disease. On vaginal speculum examination, an infected polypoid tumor protruding from a dilated cervical canal was disclosed. While the patient was under extensive antibiotic treatment, uterine curettage was performed. This was followed a few days later by a total abdominal hysterectomy and bilateral salpingo-oophorectomy. Histopathological examination demonstrated the tumor to be Mullerian adenosarcoma of the uterine endometrium. The patient had postoperative vaginal vault irradiation, and more than 1 year later she is alive with no evidence of recurrent disease. Diagnosis and management of uterine Mullerian adenosarcoma is discussed. PMID- 3031377 TI - Myasthenic syndrome (Eaton-Lambert syndrome) associated with pulmonary adenocarcinoma. AB - A case of a 57-year-old man who presented with the clinical features of Eaton Lambert syndrome preceding the diagnosis of lung adenocarcinoma at autopsy by 7 years, is reported. Although myasthenic syndrome is intimately associated with pulmonary small cell carcinoma, which connotes a grave prognosis, a small percentage of the tumor can be squamous cell carcinoma or adenocarcinoma, which may be resectable. Therefore, a continued search for evidence of intrathoracic neoplasm must be pursued following manifestations of myasthenic syndrome. PMID- 3031378 TI - Cardiac beta-adrenergic receptors in diabetic rats: alteration of guanyl nucleotide regulation. AB - Cardiac beta-adrenergic receptors (beta AR) were studied in rats after 4 months of diabetes induced by streptozotocin (50 mg/kg, i.v.). Saturation binding studies performed using [125I]-iodocyanopindolol (ICYP) as an antagonist ligand showed that the number and affinity of beta AR were not significantly altered. Analysis of competition curves of ICYP binding by the agonist (-)-isoproterenol showed that diabetes induces an increase in the (-)-isoproterenol IC50 and a steepening of the curve ("pseudo-Hill" coefficient not significantly different from 1) in the absence of Gpp(NH)p, while these parameters were not significantly modified in the presence of Gpp(NH)p (10(-4) M). These results indicates that beta AR were not significantly reduced in number but altered in their capacity to efficiently couple with the GTP-binding protein of adenylate cyclase (AC). These data can be correlated to the alteration of beta-adrenergic stimulation of cardiac AC previously observed after 4 months of diabetes. The characteristics of beta AR alteration at this stage of the disease are similar to those of a first step of a beta AR desensitization process due to overstimulation. PMID- 3031379 TI - Incubated or superfused rat lung parenchymal strip: a valid preparation for direct measurement of beta-responses. AB - Responses to the beta-adrenoceptor agonists isoprenaline (Iso) (non selective), salbutamol (Sal) (beta 2-selective) and noradrenaline (NA) (beta 1-selective) were studied on incubated and superfused rat lung strip. In both conditions, Iso and Sal elicited dose-related relaxations of lung strip and these responses were unaffected by the presence of phentolamine (Phen) 10(-5) M. Contractile responses to NA were obtained when it was used alone whereas in the presence of Phen 10(-5) M, NA elicited relaxant responses. The relative potencies of Iso : Sal : NA (plus Phen) were 100 : 20 : 0.69. This demonstrates that beta 2-adrenoceptor is the predominant beta-subtype involved in responses. Propranolol (10(-4) M) completely abolished the relaxant responses to Iso. However, after the addition of Iso tissue responded normally to drugs acting by other mechanisms. Thus, relaxations were obtained with caffeine and contractions with acetylcholine (AcH) or NA. This shows that responses to Iso are mediated only by beta-adrenoceptors. Propranolol and the selective beta-blocking agents butoxamine (beta 2) and atenolol (beta 1) competitively antagonized the effects of Iso. The Schild plots obtained had slopes which did not differ significantly from -1 and mean pA2 values were: 7.82 for propranolol; 6.32 for butoxamine and 5.22 for atenolol. These experiments were carried out in the absence of catecholamine uptake inhibitors, since in a preliminary set of experiments no significant changes of lung strip responses to Iso were observed in the presence of cocaine (10(-5) M) or corticosterone (10(-5) M). In conclusion, it appears that incubated rat lung strip possesses intrinsic tone and, although it cannot be considered a representative preparation of peripheral airways, it provides a valid model for investigating the direct effects of drugs which act at beta-adrenoceptors. PMID- 3031380 TI - Effect of clomipramine treatment on dog platelet alpha 2-adrenergic receptivity. AB - The effects of clomipramine treatment (5 mg/kg s.c. a day during 21 days) on dog platelet alpha 2-adrenoreceptivity were investigated. The radioligand binding studies on platelet membrane preparations showed that the antidepressant treatment promotes a decrease in the number of [3H]-yohimbine binding sites. The measurement of the inhibition of PGE1-induced cyclic AMP accumulation in the whole platelet indicated that the biological event immediately associated with alpha 2-adrenergic stimulation was not modified. The aggregatory response to ADP or ADP plus adrenaline was totally abolished in clomipramine-treated platelets. The diastolic pressor responses to noradrenaline or adrenaline injections (after propranolol and prazosin administration) in chloralose anaesthetized dogs were not affected by clomipramine treatment. This work shows that variations in the binding capacities of alpha 2-adrenergic sites are not always linked to modifications of related physiological events. PMID- 3031381 TI - Modulation of ligands binding to the benzodiazepine receptor by an endogenous inhibitor (GABA-modulin) and bicuculline. AB - We have prepared and purified an endogenous protein (brain neuropeptide) by extraction with 0.025 N acetic acid and precipitation with a saturated ammonium sulfate solution followed by column chromatography. This protein was isolated and partially purified (approx. 500 times). When added to Triton X-100 treated synaptic membrane preparation obtained from rat CNS, this modulates specific high affinity GABA binding site without affecting low affinity site. This protein was called GABA-modulin (GM). GM like bicuculline (competitive GABA antagonist) modulates the affinity of benzodiazepine receptor ligands. But the effect of bicuculline is more important than that of GM. Bicuculline increases or decreases respectively the binding of 3H-beta-CCM or 3H-flunitrazepam by approximately 60%. In the same conditions, at a concentration of 12 micrograms/ml GM increases or decreases respectively the binding of 3H-beta-CCM or 3H-flunitrazepam by approximately 35% and no more modulation was obtained even the concentration of GM was increased. Bicuculline and GM doesn't modify the specific binding of 3H-Ro 15-1788 to the BZD receptor. It is concluded that GM exhibit a lower ability than bicuculline to modulate the binding of the BZD receptor ligands because GM interacts only with the GABA high affinity binding site without affecting the low affinity binding site. These observations suggest that the two GABA binding sites probably interact with the BZD receptor. PMID- 3031383 TI - The in situ isolated larynx for evaluating peripheral opiate receptor antagonists. AB - The in situ isolated rodent larynx preparation can be utilized for the detection of peripheral opiate receptor antagonists. Measurements of peripheral opioid induced laryngospasm and central opioid-induced respiratory depression can be made in each preparation. Fentanyl citrate was used to stimulate both peripheral and central opiate receptors and [D-Ala2-Met5] enkephalinamide was used to stimulate only peripheral opiate receptors. Compounds that inhibit both laryngeal and respiratory effects of fentanyl, e.g., naloxone HCl, can be considered both central and peripheral opiate receptor antagonists. Compounds that inhibit only the peripheral laryngeal effects of fentanyl, e.g., naltrexone methylbromide, can be considered peripheral opiate receptor antagonists. The description of this preparation provides a simple and sensitive anesthetized animal model for the detection and characterization of compounds acting at peripheral and/or central opiate receptors. PMID- 3031382 TI - Interaction of carbolines and some GABA receptor ligands with the GABA and the benzodiazepine receptors. AB - The binding of different drugs were investigated on benzodiazepines and GABA receptors using P2 fraction or 0.05% Triton X-100 treated membrane preparation (TX-100 P) obtained from rat's CNS. Bicuculline and picrotoxin have the ability to modulate the specific 3H-flunitrazepam binding to its receptor present in P2 fraction; this modulation decreases for bicuculline and disappears for picrotoxin when a TX-100 P was used. Bicuculline at the opposite of picrotoxin displaces 3H GABA binding on TX-100 P. The beta-CCE binds to the brain benzodiazepines receptor with a high affinity (IC50 = 8.5 nM on TX-100 P) and does not inhibit the binding of 3H-GABA on TX-100 P. On the contrary, other related carbolines such as harmine, harmaline, harmane and harmalol have a much lower ability to inhibit flunitrazepam binding (IC50 between 28 and 130 microM with TX-100 P) and can also inhibit 3H-GABA binding (IC50 between 75 and 186 microM with TX-100 P). But the inhibition of 3H-GABA binding by those related carbolines can't be reversed by a benzodiazepine like flunitrazepam or a benzodiazepine antagonist like Ro 15-1788. PMID- 3031384 TI - 16th annual UCLA symposium. Abstracts: Human retroviruses, cancer and AIDS: approaches to prevention and therapy. PMID- 3031385 TI - Recognition of T-cell murine leukemia by bacteriocin (colicin); correlation with transplantation experiments. AB - The presence of potential murine leukemia and overt leukemia cells in various organs at different phases of leukemogenesis was demonstrated by transplantation experiments and sensitivity to bacteriocin (colicin HSC10). Such correlation was found in three experimental models: AKR mice developing spontaneous T-cell leukemia and BL/6 mice infected with radiation leukemia virus variants inducing a high or low overt T-cell leukemia incidence. The sensitivity to bacteriocin was evaluated by testing the cell cycle perturbation following in-vitro incubation of lymphoid cells with colicin (or Tris buffer as controls) monitored by flow cytometry. The analysis was based on measuring relative differences in fluorescence intensity of propidium iodide stained DNA in the individual cells. The interaction with colicin of leukemic cells and lymphoid cells containing potential leukemic cells (PLC) resulted in a reduction in the cell number of the G0/G1 and SG2M phases while cells accumulated in the "pre-G1" channels. In contrast, normal lymphoid cells exposed to bacteriocin did not show such changes in the DNA histograms. The distribution pattern of PLC in the thymus and spleen (in the models tested) obtained by transplantation studies coincided with sensitivity of spleen and thymus cells to colicin. However, in most instances, the PLC in the bone marrow were not recognized by colicin, but their leukemogenic potential was reduced following interaction with colicin as shown by PLC transplantation studies. It is thus suggested that colicin might be used for identification and eradication of transformed cells. PMID- 3031386 TI - Radiation leukemia virus and X-irradiation induce in C57BL/6 mice two distinct T cell neoplasms: a growth factor-dependent lymphoma and a growth factor independent lymphoma. AB - Two different classes of neoplastic T cells were isolated from radiation leukemia virus (RadLV)-inoculated and from X-ray-treated C57BL/6 mice. One consisted of growth factor-dependent T-cell lymphoma (FD-TCL) lines which were established from the spleens and thymuses of treated mice within a day of lymphoma detection. FD-TCL cells were often eudiploid and could be grown in pure culture only at high concentrations, or on stromal feeder layers. Non-thymic, factor-dependent TCL cells produced interleukin-2 upon lectin stimulation, and were autostimulatory because they secreted growth factor(s) constitutively. Single cell cloning of FD TCL cells in semisolid medium required the addition of exogenous conditioned medium. In vivo, FD-TCL cells that were injected intraperitoneally or intravenously homed to the spleen, proliferated in it and killed the injected mice. FD-TCL cells did not produce local tumors at the site of subcutaneous injection. The isolation and study of FD-TCL cells was facilitated by their cultivation on stromal hematopoietic monolayers in supplemented "lymphocyte medium", until an autostimulating, self-sustaining concentration of FD-TCL cells was obtained. FD-TCL cells could not be grown from lymphoid tissue of normal, control mice. In contrast, T-cell lymphoma (TCL) lines, which were established from virus-induced thymomas which had been kept in situ for 4-6 weeks after detection, consisted of factor-independent cells that possessed an aneuploid karyotype (in some cases trisomic for chromosome No. 15), and produced local tumors at the site of subcutaneous injection. These cells could be cloned in semisolid medium without addition of exogenous factor(s). The phenotypic markers of TCL cells differed from those of FD-TCL cells, suggesting heterogeneity in the stages of differentiation at which cells can give rise to growth factor independent (TCL) and to growth factor-dependent (FD-TCL) lines. PMID- 3031387 TI - Bone marrow phenotype determines genetic resistance to RadLV-induced leukemia in radiation chimeric mice. AB - The development of RadLV-induced T-cell leukemia is a multistep process which evolves along the bone marrow-thymus axis. This process has been shown to be under the control of resistance and susceptibility genes. The relative importance of bone marrow and thymic phenotypes in this genetic control have not been established. We have constructed radiation chimeras with bone marrow from susceptible C57BL/Ka and thymus from resistant B10.A(5R) mice (and vice versa). The rate of leukemia development in the various groups indicates that the phenotype of the bone marrow and not that of the thymus determines the expression of resistance or susceptibility. PMID- 3031388 TI - Experimental transmission of enzootic bovine leukosis to cattle, sheep and goats: infectious doses of blood and incubation period of the disease. AB - A group of 49 BLV-free recipient animals (24 cattle, 15 sheep and 10 goats) were inoculated intradermally with serial dilutions of blood collected on two BLV positive donor cows. One donor had a high lymphocytosis and high antibody titers to gpBLV antigens; these two parameters were low for the second donor. The number of lymphocytes which induced BLV infection in recipient animals varied widely with the donor. The high infectivity of a donor seemed to be correlated with high lymphocytosis and high antibody titers to gpBLV antigens. Identification and removal of infectious animals would reduce or stop the spread of the infection in a herd, and could be used in the strategy to eradicate the disease. A given inoculum can be infectious in sheep and, at the same time, harmless in cattle. The incubation periods, apparently shorter in sheep, were generally in the range from 2 to 5 weeks for the three species, and exceptionally above. PMID- 3031389 TI - Direct radioimmunoassay of angiotensin-converting enzyme in sera from patients with pulmonary diseases. PMID- 3031391 TI - Presence of Weibel-Palade bodies in lymphatic endothelium of the cat. PMID- 3031390 TI - Pulmonary alveolar proteinosis: determination of prostaglandins and leukotrienes in lavage fluid. PMID- 3031392 TI - [Treatment of herpes infections with acyclovir]. PMID- 3031393 TI - [Vasopressinergic system (II)]. PMID- 3031394 TI - [ELISA method for false positive reaction for the human immunodeficiency virus in a patient with alcoholic liver disease]. PMID- 3031395 TI - [Therapy of herpes simplex and varicella zoster infections in the ENT area]. AB - The article reports on 41 patients having infections induced by Herpes simplex and Herpes zoster virus. Systemic intravenous administration of acyclovir results in a very rapid reduction of pain and mucosal changes in herpetic stomatitis. In cutaneous lesions of the trigeminal nerve branches induced by Herpes zoster virus there is also a very rapid reduction of pain and efflurescence after 3 days. In 16 patients suffering from Ramsay Hunt syndrome, also known as Herpes zoster oticus, lesions of the facial nerve function were present. 8 Patients demonstrated cochleovestibular signs and symptoms, 2 had flat inner ear hearing loss of 40 dB, 1 reduced unilateral caloric response. Treatment was effected by intravenous administration of acyclovir and simultaneous classical symptomatic therapy consisting of intravenously administered dextrane, cortisone and antiinflammatory drugs. Symptomatic therapy is necessary because acyclovir stops the replication of viruses but does not influence the disturbed nerve function. In 2 cases with a damage of more than 90% of the facial nerve fibres, endaural decompression of the geniculate ganglion was performed. Cochleovestibular deficits improved to normal during one week and all facial lesions within 8 months. Drug-related side effects were seen in one patient who had an exanthema. PMID- 3031396 TI - [Nuclear magnetic resonance tomography: a new imaging diagnostic procedure of the petrous bone and cerebellopontile angle]. AB - Nuclear magnetic resonance imaging (NMRI) is a new technique enabling three dimensional scanning of the petrous bone without ionizing radiation. Contrary to computer tomography NMRI enables visualization of the structures of the inner ear and the cerebello-pontine angle without masking by overprojecting bony structures. The resolution of inflammatory processes and tumors is excellent. NMRI is inferior to CT in demonstrating bony destruction, and is therefore not so effective in the evaluation of glomus tumours. The use of surface coils in combination with a special gradient-zoom-technique allows high resolution imaging. This and Gadolinium-DTPA for enhancement opens new perspectives in the diagnosis of small acoustic neuromas. PMID- 3031397 TI - Site of inhibitory action of CRE 9-41 on ACTH release from isolated rat pituitary cells. AB - The corticotropin-releasing factor (CRF) analog CRF 9-41 inhibits CRF, but not forskolin or dibutyryl cyclic AMP, stimulated release of ACTH from isolated pituitary cells. CRF 9-41 also blocks CRF-stimulated accumulation of cyclic AMP in a parallel dose dependent fashion. CRF 9-41 has no effect on basal ACTH release or cAMP levels. This substantiates that the analog acts as a direct CRF antagonist and that the site of this inhibition is most likely at the level of binding of CRF to its receptor on the corticotrope. Various substances, including most prominently glucocorticoids, inhibit release of ACTH from the pituitary. In an effort to develop another class of inhibitors, Rivier et al recently synthesized analogs of corticotropin releasing factor (CRF). One among these, alpha-helical ovine CRF 9-41 blunts adrenalectomy and stress induced ACTH release in non-anesthetized rats. At micromolar concentrations, CRF 9-41, shifts rightward the dose response of isolated pituitary cells to ovine CRF. Thus, the authors suggested that CRF 9-41 acts as a competitive antagonist to CRF-induced ACTH secretion. CRF appears to act through stimulation of adenylate cyclase. To determine the potential site of action of CRF 9-41 in the activation sequence for adenylate cyclase, we studied its effects on pituitary cyclic AMP formation and ACTH secretion from dispersed anterior pituitary cells derived from normal adult rats, as well as, its interaction with cyclic nucleotide agonists. PMID- 3031398 TI - Ketotifen increases cyclic adenosine 3',5'-monophosphate in intact human lymphocyte and potentiates other adenylate cyclase activating agents. AB - We have investigated the effects of ketotifen on the cyclic adenosine 3',5' monophosphate (cyclic AMP) response of intact human lymphocyte and its interaction with adenylate cyclase activating agents. In the presence of cyclic AMP phosphodiesterase inhibitor (3-isobutyl-1-methyl-xanthine), ketotifen (10(-8) 10(-4) M) caused an 80% increase in cyclic AMP content of human lymphocyte, a magnitude similar to that observed with hydrocortisone. The cyclic AMP level peaked at about 15 minutes and remained elevated for at least 45 minutes. In addition, ketotifen (10(-6)-10(-4) M) markedly potentiated the effect of several adenylate cyclase stimulating agents, including L-isoproterenol, prostaglandin E1 and forskolin. The biochemical mechanisms underlying these effects are unknown. It may be at least partly related to the ability of ketotifen to reverse and prevent beta 2 adrenoceptor desensitization and to promote the formation of hormone - nucleotide - high affinity receptor complex. These effects may contribute to its prophylactic effect in the treatment of bronchial asthma. PMID- 3031399 TI - Direct evidence for the interaction between papaverine and alpha 2-adrenergic receptors in canine platelets. AB - The effects of papaverine on specific [3H]-yohimbine binding to canine platelet alpha 2-adrenergic receptors and on the platelet aggregation were assessed and compared with those of verapamil. Both compounds concentration-dependently inhibited [3H]-yohimbine binding with KI values for respective compounds of 0.39 +/- 0.05 microM (n = 3) and 15 +/- 0.19 microM (n = 3). In the presence of either compound KD values in Scatchard analysis of the equilibrium ligand binding increased in concentration-dependent manner, whereas Bmax did not change, indicating competitive inhibition of the ligand binding by these compounds. (-) Epinephrine (3 microM) potentiation of adenosine diphosphate (ADP, 0.1 microM) aggregation was inhibited by papaverine with IC50 of 11 +/- 3.6 microM (n = 4). In the same experiments verapamil inhibited the platelet aggregation with lower IC50 (3.1 +/- 0.87 microM, n = 4) in comparison with that for papaverine. These results suggest that papaverine, like verapamil, inhibits physiological response of canine platelets through alpha-adrenergic receptor stimulation by direct interaction with the receptors. PMID- 3031400 TI - The beta-carboline ZK 93423 inhibits reticulata neurons: an effect reversed by benzodiazepine antagonists. AB - A novel beta-carboline with benzodiazepine-like properties has recently been synthesized. We compared the effect of the i.v. administration of this drug, ZK 93423, with diazepam on the activity of nigral pars reticulata neurons which are known to be very sensitive to the inhibitory effect produced by GABA-mimetics and benzodiazepines. ZK 93423 (0.05-1.0 mg/kg) inhibited reticulata cells in a dose related manner up to the cessation of their activity. Since the maximal rate inhibition elicited by diazepam (1.0 mg/kg) was some 55% of baseline, ZK 93423 showed a much greater potency. Moreover, the firing depression by ZK 93423 was prevented and reversed by two benzodiazepine receptor antagonists: Ro15-1788 and ZK 93426. However, the dosage of Ro15-1788 required for these actions was at least five times higher than that for the blockade of the diazepam effect. The results indicate that the beta-carboline agonist ZK 93423 decreases the activity of reticulata neurons more effectively than diazepam. PMID- 3031401 TI - Effects of different light wavelengths at equal irradiance on testis weight, protein content, and K-paranitrophenylphosphatase activity in the Syrian hamster. AB - The effects of different light wavelengths at equal irradiance on testis weight, testis protein content, and testis K-paranitrophenylphosphatase (K-pNPPase) activity were studied in the Syrian hamster. One group (long photoperiod) was maintained on a light:dark cycle of 14:10, and another group (short photoperiod) on a cycle of 10:14. Five other groups were maintained on a cycle of 10:14 but with a one hour pulse of equally intense illumination in the middle of the dark period with UV, blue, green, yellow or red light. Animals exposed to a long photoperiod or UV, blue or green light pulses had significantly greater testis weights--up to eightfold greater than those in the yellow or red or short photoperiod groups. Organ protein content closely paralleled organ weight, but the protein/wet weight ratio was consistently higher in the large organ groups. K pNPPase and Mg-pNPPase activities were significantly higher in the large organ groups, even when expressed per mg protein. Therefore, at a balanced irradiance of 0.2uW/cm2, light wavelength exerts a profound effect on testicular weight, protein content, and K-pNPPase and Mg-pNPPase activities. Testicular involution is a process that is selective with regard to protein biosynthesis. PMID- 3031402 TI - Effect of bromocriptine on lipid plasma levels in rats. AB - Results show that bromocriptine induced marked alterations in plasma levels of cholesterol and lipids in response to acute and chronic administrations in rats. Two hours after an I.P. dose of 10 mg/kg, bromocriptine mesylate caused significant reductions in plasma levels of total high density lipoprotein (HDL) and high density lipoprotein cholesterol (HDL cholesterol). At a dose of 20 mg/kg, bromocriptine mesylate induced significant elevations in plasma levels of total cholesterol, total HDL, HDL cholesterol, total low density lipoproteins (LDL), and low density lipoprotein cholesterol (LDL cholesterol). Injected at a dose of 4 or 10 mg/kg daily for 14 consecutive days, bromocriptine mesylate caused significant increases in plasma levels of total cholesterol, LDL cholesterol and total LDL whereas the levels of HDL cholesterol, total HDL triglycerides (TG) were reduced. At a dose of 20 mg/kg all parameters were significantly increased. Marked hyperglycaemia was noticed in response to doses of 10, 15 and 20 mg/kg injected daily for 14 consecutive days or 2 hrs after a single administration of 15 mg/kg. Plasma insulin activity was reduced 2 hours after injection of bromocriptine at a dose of 15 mg/kg Likewise, a significant reduction in plasma insulin activity was observed in response to daily I.P. injections of bromocriptine at a dose of 15 mg/kg. Hyperglycaemic and hypoinsulinaemic effects of bromocriptine (acute and chronic) were markedly decreased when sulpiride, a dopaminergic D2 antagonist, was injected at an I.P. dose of 10 mg/kg before bromocriptine. Plasma ACTH activity was significantly increased in response to bromocriptine (15 mg/kg I.P.) in acute and chronic experiments. This effect was markedly diminished when sulpiride was injected prior to bromocriptine. In conclusion, bromocriptine induced marked elevations in plasma levels of total cholesterol and lipids which are likely to be related to hyperglycaemic and hypoinsulinaemic effects. PMID- 3031404 TI - Effect of norepinephrine on renal tubular Na-K-ATPase and oxygen consumption. AB - Previous studies from this laboratory have demonstrated that norepinephrine (NE) increases sodium dependent phenylalanine uptake by in vitro rat cortical tubules. In the present study, we have examined whether the observed increase in proximal tubular sodium transport, during NE treatment, is related to changes in membrane Na-K-ATPase and cellular oxygen consumption. Treatment of intact tubules with NE increased microsomal Na-K-ATPase activity but had no effect on cellular oxygen consumption or ouabain inhibitable oxygen consumption. The increased Na-K-ATPase activity is consistent with the observed increase in sodium transport while the lack of a detectable effect on oxygen consumption suggests that the increased transport does not require additional oxygen utilization. PMID- 3031403 TI - Coexistence of central and peripheral benzodiazepine binding sites in the human pineal gland. AB - The pineal gland and particularly its major hormone, melatonin, may participate in several physiological functions, including sleep promotion, anticonvulsant activity and the modulation of biological rhythms and affective disorders. These effects may be related to an interaction with benzodiazepine receptors, which have been demonstrated to be present in the pineal gland of several species including man. The present study examined the characteristics of benzodiazepine binding site subtypes in the human pineal gland, using [3H]flunitrazepam and [3H]PK 11195 as specific ligands for central and peripheral type benzodiazepine binding sites respectively. Scatchard analysis of [3H]flunitrazepam binding to pineal membrane preparations was linear, indicating the presence of a single population of sites. Clonazepam and RO 15-1788, which have a high affinity for central benzodiazepine binding sites, were potent competitors for [3H]flunitrazepam binding in the human pineal, whereas RO 5-4864 had a low affinity for these sites. Analyses of [3H]PK 11195 binding to pineal membranes also revealed the presence of a single population of sites. RO 5-4864, a specific ligand for peripheral benzodiazepine binding sites was the most potent of the drugs tested in displacing [3H]PK 11195, whereas clonazepam and RO 15-1788 were weak inhibitors of [3H]PK 11195 binding to pineal membranes. Overall, these results demonstrate, for the first time, the coexistence of peripheral and central benzodiazepine binding sites in the human pineal gland. PMID- 3031405 TI - Organ and tumor distribution of (18F)-2-fluoro-2-deoxy-D-glucose in fasting and non-fasting rats. AB - Differences in the uptake of (18F)-2-fluoro-2-deoxy-D-glucose ((18F)FDG) in various normal organs and the Rous sarcoma of fasted and unfasted rats were studied at 5, 15, 30, 60, and 120 minutes after i.v. injection. The uptake of (18F)FDG in the tumor, spleen, and testis increased for 120 minutes, while uptake in the other organs was either level (brain, heart, white fat) or cleared off. The uptake was higher in the tumor than in the normal organs. The fraction of viable tumor tissue as measured morphometrically correlated intraindividually with the uptake of (18F)FDG--an increase of 1% of vital tumor corresponded to a 1.01-fold increase in tumor uptake of (18F)FDG. The nutritional state was of importance for the uptake of (18F)FDG into the heart, testis and brown fat. (18F)FDG is taken quantitatively up by the viable parts of the Rous tumor; this may make it possible to follow the response of treatment in individual tumors also in man with (18F)FDG and positron emission tomography (PET). PMID- 3031406 TI - Diurnal rhythm of plasma immunoreactive corticotropin-releasing factor in normal subjects. AB - Plasma corticotropin-releasing factor (CRF), corticotropin (ACTH) and cortisol levels were simultaneously determined by radioimmunoassays at 0600 h, 1200 h, 1800 h and 2200 h in six normal subjects, in order to examine whether the diurnal rhythm in plasma CRF exists and how it correlates to the diurnal rhythm in plasma ACTH and cortisol concentration. The highest CRF level was observed at 0600 h (7.0 +/- 1.2 pg/ml) and significantly lower levels (p less than 0.01) at 1800 h (1.7 +/- 0.2 pg/ml) and 2200 h (1.9 +/- 0.4 pg/ml). A clear diurnal rhythm was demonstrated in plasma ACTH and cortisol levels, with the highest values at 0600 h (44.6 +/- 8.1 pg/ml and 15.9 +/- 2.0 micrograms/dl, respectively) and the lowest at 2200 h (12.3 +/- 2.8 pg/ml and 4.6 +/- 1.0 micrograms/ml, respectively). These results suggest that the diurnal rhythm in ACTH and cortisol is under the regulation, at least in part, of the diurnal rhythm in CRF secretion. PMID- 3031407 TI - Binding sites for atrial natriuretic factor (ANF) in kidneys and adrenal glands of spontaneously hypertensive (SHR) rats. AB - Binding sites for atrial natriuretic factor (ANF) were studied in kidneys and adrenal glands of 17 week old male spontaneously hypertensive (SHR) and Wistar Kyoto (WKY) normotensive rats by quantitative autoradiography using 125I-ANF-28. In kidney, 125I-ANF-28 binding sites were found in high concentrations in glomeruli and in much lower concentrations in the renal papilla. In adrenal gland, 125I-ANF-28 binding sites were highly localized to the zona glomerulosa and were of moderate density in the inner cortical regions. ANF binding sites did not occur in the adrenal medulla. The maximum binding capacity (Bmax) of 125I-ANF 28 was reduced by 50% in the kidney glomeruli of SHRs compared to WKY controls. In contrast, the affinity constant (Ka) for 125I-ANF-28 was elevated by 100% in kidney glomeruli of SHRs. There were no significant strain differences in values for Bmax or Ka for 125I-ANF-28 binding in the adrenal zona glomerulosa. These findings suggest that the natriuretic and diuretic actions of ANF within kidney glomeruli may be compromised in adult SHR rats and these alterations may contribute to the development and maintenance of hypertension in rats of this strain. PMID- 3031408 TI - Beta-endorphin and ACTH in plasma: effects of physical and psychological stress. AB - The effects of the expectancy of an official race (22000 m) and the performance of this last event on plasma levels of beta-Endorphin (B-End) and ACTH have been assessed. In a group of nine athletes, samples were obtained first in basal conditions; second in the day of the run before the warming up period and third after running. B-End immunoactivity was increased from 15.7 +/- 2.0 pg/ml to 23.4 +/- 2.5 pg/ml before the run and up to 30.6 +/- 2.9 pg/ml after the trial. ACTH levels were increased from 8.4 +/- 1.2 pg/ml to 17.9 +/- 2.3 pg/ml before running and up to 36.2 +/- 3.9 pg/ml after running. The results suggest that psychological and physical stress act synergically to increase the levels of B End and ACTH during the practice of physical exercise. PMID- 3031409 TI - Atrial natriuretic peptide binding properties of purified rat glomerular membranes. AB - 125I-ANP (3-[125I] iodotyrosyl28) binding studies with purified rat glomerular membranes indicate two types of physiologically relevant hormonal receptors, Types I and II, Kd approximately 5 pM and approximately 2.5 nM, respectively. All preparations were essentially free of capsular and tubular contamination. Binding data indicated that Type I receptors were three times more concentrated than Type II receptors in purified membrane fractions. When purified membranes were cross linked with 125I-rANP, using disuccinimidyl suberate and separated by SDS-PAGE, approximately 75- and approximately 140-kDa proteins were specifically labeled in a ratio of approximately 3:1, respectively. Thus, in purified renal glomerular membranes, Type I receptors with molecular weight of approximately 75-kDa appeared to predominate and would be detectably saturated at circulating ANP concentrations as low as 15 pg/ml. These findings could account for the exquisite sensitivity of natriuretic response to ANP. PMID- 3031410 TI - Cholesteryl esterase activities in ventricles, isolated heart cells and aorta of the rat. AB - Cholesteryl esterase activities were determined in homogenates of rat heart (ventricles), isolated, calcium-tolerant, cardiac myocytes and aortic tissue and were compared with acid and neutral triglyceride lipase activities in these fractions. Using cholesteryl oleate/phosphatidylcholine/taurocholate emulsions and digitonin pretreatment of the enzyme fractions, acid and neutral cholesteryl esterase activities were measured in all tissue preparations. In contrast to the acid and neutral triglyceridase and acid cholesteryl esterase activity, the neutral cholesteryl esterase activity was subject to substrate inhibition. Upon isolation of cardiac myocytes, and in contrast with the recovery of neutral triglyceride lipase activity, only a small portion of the neutral cholesteryl esterase (6%) was recovered, suggesting that nonmyocyte neutral cholesteryl esterase activity markedly contributes to the relatively high activity detectable in whole ventricular homogenates. The recovery of large amounts of neutral cholesteryl esterase activity in the supernatant of collagenase-digested heart tissue, obtained during the isolation of myocytes, which is also markedly enriched in activities of two endothelial marker enzymes (5'-nucleotidase and angiotensine-converting enzyme) may indicate the predominant contribution of neutral cholesteryl esterase activity from coronary endothelial cells to this activity detectable in ventricular homogenates. Relative to the activity in ventricular and myocyte homogenates, aorta homogenates possessed the highest specific neutral cholesteryl esterase activity. We propose that in addition to coronary endothelium, smooth muscle cells also contribute to the neutral cholesteryl esterase activity in ventricular homogenates.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3031412 TI - Diet and Crohn's disease. PMID- 3031413 TI - Dietary fibre--definition, chemistry and analytical determination. AB - Dietary fibre includes non-starch polysaccharides and lignin that are not digested or absorbed in the human small intestine. It contains a mixture of chemically complex polysaccharides. Lignin is a highly cross-linked complex polymer of phenylpropane units. The plant cell wall is the main source of dietary fibre and its structure is reviewed briefly. The structure of the main dietary fibre polysaccharides is then summarized. The demarcation between starch--the main digestible polysaccharide--and dietary fibre presents some problems due to more or less enzyme resistant starch fractions that occur naturally or are formed with processing. "Resistant starch" formed during baking passes through the small intestine in the rat and, probably, in man and is fermented in the colon. It should therefore also be regarded as dietary fibre. Methods for dietary fibre determination fall into two categories: gravimetric methods, weighing the dietary fibre after removal of other components; component analysis methods, determining monomeric composition of fibre polysaccharides (preferably by gas-liquid chromatography) supplemented with a gravimetric lignin determination and separate assay of uronic acid components (pectin). Recently developed enzymatic gravimetric methods are most convenient for the assay of total dietary fibre or water soluble and insoluble fibre separately, whereas component analysis is required for determining the dietary fibre composition. PMID- 3031411 TI - The effect of mengovirus infection on lipid synthesis in cultured Ehrlich ascites tumor cells. AB - The concept of generally increased lipid synthesis during the initial 2/3 of picornaviral infectious cycles, held by several authors, needs differentiation. In mengovirus-infected Ehrlich ascites tumor cells, an increase in the rate of synthesis of phosphatidylcholine could be confirmed, but for phosphatidylethanolamine constant to decreasing rates of synthesis were found. Moreover, phosphatidylinositol was increasingly synthesized in the midst of the infectious cycle. The changes observed might have their functional expression in the proliferation of smooth cytoplasmic membrane systems that provide the structural framework for the replication of picornaviral RNA and virus assembly. The alterations in the labeling patterns of phosphatidylinositol, phosphatidylglycerol and diphosphatidylglycerol late in virus infection point to increased turnover of these compounds, possibly mediated by phospholipase D. The formation of lysophosphatidylcholine (cytolytic effect) and bis(monoacylglyceryl)phosphate in the final phase of the infectious cycle might be correlated with the liberation of lysosomal enzymes and the development of the cytopathic effect. PMID- 3031414 TI - Physiological properties of dietary fibre. PMID- 3031415 TI - Dietary fibre and the colon. PMID- 3031416 TI - Dietary fibre and mineral metabolism. AB - Current dietary recommendations urge, inter alia, an increased consumption of fibre-containing foods. Some experimental studies made on various animals and man indicate that the associated increases in intakes of fibre and phytic acid may prejudice mineral status respecting calcium, magnesium, iron and zinc. An examination has been made of the experimental evidence, also of the epidemiological evidence on numerous types of populations, past and present, developing and developed. It has been concluded that diets high in fibre, characteristically do not have meaningful ill effects on well-being or unequivocally enhance morbidity. In particular populations in certain regions where deleterious effects have been reported it is judged that local factors, not wholly understood, are in operation. In assessing the extent of the benefit to be derived from the dietary changes urged, results must be viewed holistically and not in isolation. It is believed that the beneficial effects respecting reduced pronenesses to various degenerative diseases are likely to far outweigh the possible adverse effects of reduced bioavailability of mineral nutrients. PMID- 3031418 TI - Cholesterol-rich gallstones. PMID- 3031417 TI - The development of the concept of dietary fibre. PMID- 3031420 TI - Viral STDs: herpes simplex and human papillomavirus. PMID- 3031419 TI - Dietary fibre, carbohydrate metabolism and diabetes. PMID- 3031421 TI - [Scintigraphic functional research on normal salivary glands]. AB - Modification of the method of sialoscintigraphy was proposed. The determination of concentration of an increment of radioactivity of the glands after functional exercise was new. Scintigraphic indices of function in 90 patients with the unchanged salivary glands were presented. PMID- 3031422 TI - [Assessment of kidney function with 99mTc-pyrophosphate during the scintigraphy of the joints in rheumatoid arthritis patients]. AB - Diagnostic potentialities of investigation of the kidneys using 99mTc pyrophosphate were studied in 34 patients with rheumatoid arthritis (RA) for articular scintigraphy. The results obtained were compared with those of radionuclide renography with 131I-hippuran and excretory urography. A possibility of investigation of the kidneys with 99mTc-pyrophosphate for the assessment of parenchyma function and kidney urodynamics was shown. Taking into account a high prevalence of nephropathy in RA patients, dynamic renal scintigraphy and clearance registration were recommended at the first stage of articular scintigraphy with 99mTc-pyrophosphate in order to obtain information on the anatomotopographic site and accumulation-evacuatory function of the kidneys. PMID- 3031423 TI - Effects of nitroxides on the magnetic field and temperature dependence of 1/T1 of solvent water protons. AB - We report a study of the longitudinal NMRD profiles (proton longitudinal relaxation rates as a function of field strength) over a broad range of magnetic field (0.01 to 50 MHz proton Larmor frequency) and temperature (-9.6 to 37 degrees C) for aqueous solutions of (i) a fatty acid-nitroxide/albumin complex and (ii) 10 low molecular weight nitroxides. Analysis of the NMRD profile for the fatty acid-nitroxide/albumin complex provides a lower bound estimate for the rotational correlation time of the complex, which permits the calculation of an upper bound on the inner sphere contribution to relaxation of the uncomplexed nitroxides. Inner sphere processes, ostensibly due to water molecules hydrogen bonded to the nitroxide moiety, dominate the relaxation effects of the slowly rotating macromolecular nitroxide/albumin complex. By extrapolation, the contribution of these inner sphere processes are negligible for rapidly tumbling nitroxides free in solution, which affect solvent proton relaxation almost entirely through outer sphere processes (i.e., translational diffusion). A comparison of the relaxation data for aqueous solutions of the uncomplexed nitroxides with the theory of outer sphere relaxation of J.H. Freed (J. Chem. Phys. 68, 4034 (1978] yields values for the distance of closest approach of the water and nitroxide molecules, as well as for their relative diffusion constants, at five different temperatures. Our results indicate that the rather modest relaxivities of aqueous solutions of nitroxides increase substantially with increased solvent viscosity and with protein binding, supporting the potential utility of nitroxides for enhancement of contrast in nuclear magnetic resonance images. PMID- 3031424 TI - The role of pump number and intracellular sodium and potassium in determining Na,K pump activity in human erythrocytes. AB - The factors that determine the activity of the Na,K pump in vivo were investigated by measuring Na,K pump activity under in vivo conditions in human red cells and relating it to the intracellular content of sodium ([Na]i) and potassium ([K]i) and the number of pump units per cell (pump number). Na,K pump activity was measured as ouabain-sensitive K+ influx, pump number was determined from the maximal binding of 3H-ouabain to intact cells, and [Na]i and [K]i were measured by atomic absorption spectrophotometry in washed, packed cells. In the 81 samples studied, pump activity per cell was significantly correlated with pump number (r = .64, P less than 0.001), but was negatively correlated with [Na]i (r = -.28, P less than 0.02) and was not correlated with [K]i. An inverse relationship was found between pump number and [Na]i. When pump activity was expressed as activity per pump unit, rather than per cell, a significant relationship was seen between pump activity and [Na]i (r = .50, P less than 0.001), and a negative correlation existed between the activity per pump unit and [K]i (r = -.29, P less than 0.01). The effect of intracellular Na+ at physiologic levels on pump activity was not strong, with the activity per pump unit increasing only 25% with a doubling of [Na]i. These results indicate that pump number is the major determinant of pump activity in human red cells in vivo, while [Na]i and [K]i are of secondary importance.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3031425 TI - Increase in S-100b protein content in thyroid carcinoma. AB - S-100b protein was detectable in the soluble fraction of thyroid tissue. The concentration of S-100b protein in thyroid carcinoma tissue was three to five times higher than in normal thyroid tissue and thyroid adenoma. It is, however, not higher in the thyroid tissue of Graves' disease. The increase of S-100b protein concentration was not remarkable in carcinomatous tissue of the stomach and other digestive organs. The calmodulin content in the thyroid carcinoma tissue increased but the increment was low compared to that of S-100b protein. These data suggest that S-100b protein may play a significant role in cell maturation or differentiation. PMID- 3031427 TI - Organization and nucleotide sequence of genes encoding core components of the phycobilisomes from Synechococcus 6301. AB - Cyanobacteria possess specialized organelles, called phycobilisomes, which collect and transfer light energy to the reaction centres of photosystem II, in the photosynthetic membrane. Phycobilisomes consist of a central core, mainly composed of allophycocyanin, from which six rods radiate. We report here the isolation, for the first time, of three genes that encode core components of cyanobacterial phycobilisomes. The genes coding for the alpha- and beta-subunit apoproteins of allophycocyanin (apcA and apcB) were cloned from Synechococcus PCC 6301 and subjected to nucleotide sequence analysis. Dowstream of apcB, we found a third open reading frame (apcC) which, by comparison with known amino acid sequences, was assigned to L7.8c, a linker polypeptide associated with phycobiliproteins within the core of the phycobilisomes. Homologies between amino acid sequences deduced from the nucleotide sequence of the Synechococcus PCC 6301 apc genes and the amino acid sequences published for corresponding proteins either from cyanobacteria or chloroplast-like organelles of eukaryotic organisms, are 75% or more. The genetic organization of this photosynthetic gene cluster relative to that observed in the cyanelle genome of the flagellate Cyanophora paradoxa is discussed. PMID- 3031426 TI - Characterization of the genes encoding the haemolytic toxin and the mosquitocidal delta-endotoxin of Bacillus thuringiensis israelensis. AB - The crystalline parasporal inclusions (crystals) of Bacillus thuringiensis israelensis (Bti), which are specifically toxic to mosquito and black fly larvae, contain three main polypeptides of 28 kDa, 68 kDa and 130 kDa. The genes encoding the 28 kDa protein and the 130 kDa protein have been cloned from a large plasmid of Bti. Escherichia-coli recombinant clones containing the 130 kDa protein gene were highly active against larvae of Aedes aegypti and Culex pipiens, while B. subtilis recombinant cells containing the 28 kDa protein gene were haemolytic for sheep red blood cells. A fragment of the Bti plasmid which is partially homologous to the 130 kDa protein gene was also isolated; it probably corresponds to part of a second type of mosquitocidal toxin gene. Furthermore, restriction enzyme analysis suggested that the 130 kDa protein gene is located on the same Bti EcoRI fragment as another kind of Bti mosquitocidal protein gene cloned by Thorne et al. (1986). Hybridization experiments conducted with the 28 kDa protein gene and the 130 kDa protein gene showed that these two Bti genes are probably present in the plasmid DNA of B. thuringiensis subsp. morrisoni (PG14), which is also highly active against mosquito larvae. PMID- 3031428 TI - Nucleotide sequence of ompV, the gene for a major Vibrio cholerae outer membrane protein. AB - The nucleotide sequence of the ompV gene of Vibrio cholerae was determined. The product of the gene is a 28,000 dalton protein which, after the removal of a 19 amino acid signal sequence, produces a mature outer membrane protein of 26,000 daltons. The cleavage site was determined by amino-terminal amino acid sequencing of the purified mature protein. The DNA upstream of the gene shows the presence of a typical promoter region as judged from the Escherichia coli consensus information; however, the Shine-Dalgarno sequence is associated with a region capable of forming a secondary structure in the mRNA. The formation of this structure would inhibit binding of the mRNA to the ribosome and reduce translation. It is proposed that this structure is recognized by a positive activator in V. cholerae and because of its absence in E. coli ompV is poorly expressed. The distribution of rare codons within ompV suggests that they may serve to slow down the translation of particular domains such that the nascent polypeptide has an opportunity to take up its conformation without interference from the later formed regions. Such a mechanism could aid localization of the protein if export were by a contranslational secretion system. PMID- 3031429 TI - The gene for Escherichia coli diadenosine tetraphosphatase is located immediately clockwise to folA and forms an operon with ksgA. AB - The DNA sequences of the diadenosine tetraphosphatase gene (apaH) and of the flanking regions were determined. Three other genes were identified in the flanking regions: ksgA, apaG and folA encoding, respectively, a 16 S rRNA methyltransferase, an unidentified protein of Mr 13,826 and dihydrofolate reductase, with the order folA-apaH-apaG-ksgA. The apaH gene is thus located between folA and ksgA at 1 min on the Escherichia coli chromosome linkage map and folA is transcribed clockwise, whereas ksgA, apaG and apaH are transcribed in the opposite direction. It was shown that ksgA, apaG and apaH can be expressed from a polycistronic mRNA originating from a promoter (p1) located upstream of ksgA. However, another promoter (p2) was found within the ksgA structural gene. This promoter, active in vivo, can account for p1-independent expression of the two distal cistrons, apaG and apaH. Finally, the effect on diadenosine tetraphosphatase over-production of a frameshift mutation causing premature translational termination of apaG suggests that expression of apaG and apaH is coupled at the translational level. PMID- 3031430 TI - Physical mapping of LP51 and LP52 prophages of lysogenic strains of Bacillus licheniformis. AB - The physical maps of the LP51 and LP52 prophages in lysogenic strains of Bacillus licheniformis were constructed on the basis of data obtained by hybridization of phage DNA probes with Southern blots of restricted DNA of the lysogens. The data were compatible with the Campbell model for chromosomal integration; the attP site was mapped at 58.7-61.8 map units of the genomes of both phages. Identification of prophage-host DNA junction fragments indicated the presence of a unique attB site on the bacterial chromosome; the set of junction fragments in the strain B. licheniformis ATCC 10716 was identical to that of ATCC 11946, but different from ATCC 8187. Both the LP51 and LP52 phages used the same integration sites. Upon reinfection with either phage, the cured strains UM12 and UM18 (i.e. 10716 and 11946 cured of LP52 or LP51, respectively) turned out to be integration deficient. In surface cultures the reinfected bacteria could be maintained in the lysogenic state without, however, integrating the phage genome; when these bacteria were passaged in submerged cultures, several modes of anomalous integration were observed, and the phage segregated into a variety of forms, discernible by virulence and plaque morphology. In liquid cultures of UM12(LP51) or UM12(LP52) lytic forms finally predominated, while most lysogenized UM18 were converted into defective lysogens which contained a defective prophage in a stably integrated form. PMID- 3031431 TI - Strains overproducing tRNA for histidine. AB - Hybridization analysis of total genomic DNA indicated that Escherichia coli K12 contains a single copy of the gene encoding the histidine-accepting tRNA. This gene was subcloned onto an inducible expression vector under the control of the tac promoter. Strains carrying the resulting plasmid showed five- to six-fold increased histidine-accepting activity after induction. This overproduction of tRNAHis did not effect the growth rate of the strain or lead to derepression of the histidine biosynthetic enzymes. Neither did it have an effect on mistranslation elicited by histidine starvation. However, in cells starved for histidine by the addition of alpha-methyl histidine, the overproduction of tRNAHis interfered with the ability of the cells to recover from starvation. PMID- 3031434 TI - Homologies between small nuclear RNAs and LAV/HTLV-III sequences and their possible role in the cytopathic effect of the virus. AB - The mechanism of action of LAV/HTLV-III resulting in a pronounced cytopathic effect is still unknown. We demonstrate that the long terminal repeat (LTR) sequence and env gene of LAV/HTLV-III contain regions of 70-80% homology and/or complementarity with regions of small nuclear (sn) RNAs U1 and U2. On this basis, we hypothesize that the cytotoxic effect of LAV/HTLV-III is due - at least partly - to the inhibition of processing of cellular hnRNAs by competing for splice junction sites and/or by complexing with U1 and especially U2 RNAs. PMID- 3031432 TI - Analysis of Tn7 transposition. AB - Five gene products required for Tn7 transposition were identified using genetic complementation tests. Four of these (tnsA, tnsB, tnsC and tnsD) are essential for insertion into the attachment site, whereas the fifth (tnsE) is required for transposition to plasmids which lack this site. tnsD is not required for transposition to plasmids lacking the attachment site. This analysis used a chloramphenicol-resistant 'mini' transposon containing Tn7 termini but no complete Tn7 gene product and several compatible expression vectors containing cloned Tn7 fragments. The use of transcriptional and translational fusions allowed the identification of two promoters (P1 and P2) at the righthand end of the transposon and indicated that at least tnsA, tnsB and tnsC are translated in the same direction. Expression from P1 appears to be repressed by tnsB. PMID- 3031433 TI - DNA inversion mediated by the r-determinant of plasmid NR1: evidence for the intramolecular replicative transposition of a 23 kb IS1-flanked transposon? AB - The r-determinant (r-det) of the R plasmid NR1-Basel is a 23 kb, IS1-flanked transposon, called Tn2671, which has been shown to transpose to the genome of bacteriophage P7. Among the derivatives of phage P7::r-det we found one which carried two copies of the r-det as inverted repeats and which also contained the P7 genome segment between them in inverted orientation. Its generation is best explained by assuming that the entire 23 kb Tn2671 transposon has undergone intramolecular replicative transposition. PMID- 3031436 TI - Homologous physiological effects of nutritional antioxidants and eicosapentaenoic acid. AB - Dietary fish oils rich in eicosapentaenoic acid (EPA) possess immunostimulant, anti-inflammatory, cancer-retardant, antithrombotic and ischemia-protective properties that are remarkably parallel to the effects of high-dose nutritional antioxidants. EPA and nutritional antioxidants may show complementary activities in a wide range of preventive and therapeutic applications. PMID- 3031435 TI - Adrenergic receptor hyperactivity--a cause for pseudopheochromocytoma? AB - Two patients with classical signs and symptoms of pheochromocytoma, are described. In these two patients the 24-hr urinary excretion of dopamine, norepinephrine, epinephrine and their metabolites were normal or low, both in basal states or after attacks (spontaneous or provoked). However, the 24-hr urinary excretion of cyclic AMP was exceedingly high in one patient (13000 nmol/gm creatinine), and elevated in the second (5920 nmol/gm creatinine). Both patients were benefited by treatment with a combination of alpha and beta adrenergic blocking drugs. The first patient, after stopping treatment, developed hypertensive crisis and died. Post-mortem examination did not detect a pheochromocytoma or any other abnormality which could explain the excessively elevated cyclic AMP. In the second patient, extensive X-ray and CT examinations were negative for pheochromocytoma. A hypothesis is developed linking the symptoms and biochemical findings of both patients to an abnormality of the adrenergic receptor. PMID- 3031437 TI - Fat embolization and pulmonary infiltrates after bone marrow transplantation. AB - Pulmonary interstitial infiltrates developed in a 22-year-old female after bone marrow transplantation (BMT) for acute lymphoblastic leukemia (ALL) in second remission. She was receiving prednisone for graft versus host disease (GvH). There was some evidence of cardiac failure, but the primary diagnosis was that of cytomegalovirus (CMV) pneumonia, which resolved. Recurrent infiltrates were associated with the appearance of fat emboli in the pulmonary capillaries. There was little histological evidence of CMV pneumonitis, although other tests confirmed persistent infection. The patient recovered after further treatment directed at CMV infection and cardiac failure with a modest reduction in steroid dose. Most previous descriptions of pulmonary fat embolization (PFE) in immunocompromised patients have been derived from autopsy studies, and the majority of patients have received steroid therapy. The present case illustrates that PFE may complicate or contribute to the picture of interstitial pneumonitis (IPN) in the BMT recipient and that this syndrome may be reversible. PMID- 3031438 TI - Bilateral anaplastic Wilms' tumors: change in ploidy following treatment. PMID- 3031439 TI - A review of 1H nuclear magnetic resonance relaxation in pathology: are T1 and T2 diagnostic? AB - The longitudinal (T1) and transverse (T2) proton (1H) nuclear magnetic resonance (NMR) relaxation times of pathological human and animal tissues in the frequency range 1-100 MHz are archived, reviewed, and analyzed as a function of tissue of origin, NMR frequency, temperature, species, and in vivo versus in vitro status. T1 data from specific disease states of the bone, brain, breast, kidney, liver, muscle, pancreas, and spleen can be characterized by simple dispersions of the form T1 = AvB in the range 1-100 MHz with A and B empirically determined pathology-dependent constants. Pathological tissue T2 values are essentially independent of NMR frequency. Raw relaxation data, best-fit T1 parameters A and B, and the mean T2 values, are tabulated along with standard deviations and sample size to establish the normal range of pathological tissue relaxation times applicable to NMR imaging or in vitro NMR examination. Statistical analysis of relaxation data, assumed independent, reveals that most tumor and edematous tissue T1 values and some breast, liver, and muscle tumor T2 values are significantly elevated (p greater than or equal to 0.95) relative to normal, but do not differ significantly from other tumors and pathologies. Statistically significant abnormalities in the T1 values of some brain, breast, and lung tumors, and most pathological tissue T2 values could not, however, be demonstrated in the presence of large statistical errors. Both T1 and T2 in uninvolved tissue from tumor-bearing animals or organs do not demonstrate statistically significant differences from normal when considered as a group, suggesting no appreciable systemic effects associated with the presence of tumors compared to the statistical uncertainty. Statistical prediction analysis for both T1 and T2 indicates that of all the tissues studied, only liver hepatoma can be reliably distinguished from normal liver based on a single T1 measurement (p greater than or equal to 0.95) given the scatter in the current published data. Indeed, data scatter, not easily attributable to temperature, species, in vivo versus in vitro status, the inclusion of implanted or chemical induced tumors, or the possible existence of multiple component relaxation, is recognized as the major factor inhibiting the diagnostic utility of quantitative NMR relaxation measurements. Malignancy indexes that combine T1 and T2 data as a diagnostic indicator suffer similar problems of uncertainty. The literature review reveals a dearth of information on the temperature and frequency dependence of pathological tissue relaxation and the possible existence of multiple relaxation components.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3031440 TI - [Clinicopathologic conference. Cholestasis syndrome in epithelioid cell granuloma of the liver: diagnostic and therapeutic difficulties]. PMID- 3031441 TI - [Nutrition and carcinogenesis]. PMID- 3031443 TI - Classification system for human immunodeficiency virus (HIV) infection in children under 13 years of age. PMID- 3031444 TI - Electronic determinants of the anti-inflammatory action of benzoic and salicylic acids. AB - Ab initio, quantum chemical methods have been used to study the possible modes of binding of benzoic and salicylic acids to cyclooxygenase which lead to their anti inflammatory action. The biological data for this work were obtained from full dose response curves of the inhibitory potency of active compounds on prostaglandin production in mouse macrophages. With the help of simple regression analysis the most important reactivity indices are identified and the ionization state of the active species is discussed. From the physical significance implied by these regressions and an analysis of the electronic charge distributions of the frontier orbitals, a two-way charge transfer model is proposed. The electrostatic potentials of active and inactive congeners have been analyzed, and it is shown that an electrostatic orientation effect seems to make an important contribution to the binding of the active molecules to their receptor site. An electrostatic potential model of the binding site is proposed, and it is shown that this model is able to rationalize the source of activity or inactivity of the investigated substances. PMID- 3031445 TI - A proposed mechanism for the selective inhibition of human cytomegalovirus replication by 1-(2'-deoxy-2'-fluoro-beta-D-arabinofuranosyl)-5-fluorouracil. AB - The biological activities of 1-(2'-deoxy-2'-fluoro-beta-D-arabinofuranosyl)-5 fluorouracil (2'F-ara-FU), 1-(3'-deoxy-3'-fluoro-beta-D-arabinofuranosyl)-5 fluorouracil (3'F-ara-FU) and 1-beta-D-arabinofuranosylthymine (ara-T) were compared in human cytomegalovirus (HCMV)-infected and noninfected human fibroblasts. 2'F-ara-FU inhibited HCMV plaque formation (ED50, 16 microM for AD 169 strain) at lower concentrations than 3'F-ara-FU (ED50, 100 microM for AD 169). These nucleoside analogues are expected to be phosphorylated to their 5' phosphate forms by cellular thymidine kinase in HCMV-infected cells. The thymidine kinase activities in the virus-infected and noninfected cells were compared. Cellular thymidine kinase was increased in the virus-infected cells and showed better phosphorylation of 2'F-ara-FU than did 3'F-ara-FU. HCMV DNA polymerase was purified using affinity column chromatography, and the inhibitory effect of the 5'-triphosphate derivatives of 2'F-ara-FU (2'F-ara-FUTP) and 3'F ara-FU (3'F-ara-FUTP) against viral and host DNA polymerase alpha was examined. No significant difference in the effectiveness of inhibition was observed between viral DNA polymerase and host polymerase alpha. However, viral polymerase incorporated 2'F-ara-FUTP into newly synthesized DNA, whereas polymerase alpha did not utilize 2'F-ara-FUTP as a substrate. Thus, viral polymerase differs from host polymerase alpha in its recognition and utilization of 2'F-ara-FUTP. This difference may be important to the design of selective antiviral agents for HCMV. PMID- 3031446 TI - Ontogeny of adenosine 3',5'-monophosphate metabolism in guinea pig cerebral cortex. I. Development of responses to histamine, norepinephrine and adenosine. AB - The effects of norepinephrine, histamine and adenosine, singly or in combinations, on the accumulation of adenosine 3',5'-monophosphate were examined in slices of cerebral cortex from strain 2 guinea pigs at 40 to 68 days of gestation. The response to histamine was 2-fold at 40 days, increased to 19-fold at 55 days and declined there after toward the adult value of 4-fold. The response to adenosine was first apparent at 44 days and developed rapidly to a maximum of about 40-fold at 55 days. The response to norepinephrine remained at about 2-fold throughout the entire period. Synergistic responses to combinations of pairs of agents all became visible at 42 days and the degree of synergism was maximal by 47 to 48 days of gestation. The pharmacological characteristics of responses in fetal tissue resembled those in adult tissue in that the effects of norepinephrine in the presence of either adenosine or histamine were mediated principally by alpha-adrenergic receptors and the responses to histamine were more effectively inhibited by H1 and H2 antagonists in the presence and absence of adenosine, respectively. PMID- 3031447 TI - Ontogeny of adenosine 3',5'-monophosphate metabolism in rabbit cerebral cortex. Development of responses to histamine, norepinephrine, adenosine, and glutamate. AB - The effects of norepinephrine (NE), histamine (HIST), glutamate, and adenosine, singly and in combinations, on the accumulation of adenosine 3',5'-monophosphate (cyclic AMP) in slices of rabbit cerebral cortex were examined using tissue from animals 4 days before to 38 days after birth. A response to NE became visible 2 days before birth and increased to a maximum at 7 days after birth before declining toward the small adult value during the second post-natal week. During this period NE was at least twice as efficacious as isoproterenol, and both alpha and beta-adrenergic antagonists had prominent inhibitory effects. Responses to HIST were already apparent 4 days before birth and increased in an irregular fashion thereafter, sometimes exceeding the adult value during the second post natal week. The response to adenosine was not visible until birth and gradually increased toward the adult value during the entire period examined. Synergistic responses to various combinations of the three agents were first detected at 2 to 4 days before birth. The degree of synergism was larger during the neonatal period than that found in adult tissue; no synergism between HIST and adenosine persisted in the adult. During the first post-natal week, L-glutamate produced very large increases of cyclic AMP accumulation in the presence of either adenosine or histamine plus theophylline; smaller but substantial responses occurred in combination with NE. Responses to glutamate declined progressively after about the tenth post-natal day. PMID- 3031448 TI - Autonomous replicating sequences from mouse cells which can replicate in mouse cells in vivo and in vitro. AB - We have already reported that the cloned mouse DNA fragment (pMU65) could replicate in a simian virus 40 T antigen-dependent system in vivo and in vitro (H. Ariga, Z. Tsuchihashi, M. Naruto, and M. Yamada, Mol. Cell. Biol. 5:563-568, 1985). The plasmid p65-tk, containing the thymidine kinase (tk) gene of herpes simplex virus and the BglII-EcoRI region of pMU65 homologous to the simian virus 40 origin of DNA replication, was constructed. The p65-tk persisted episomally in tk+ transformants after the transfection of p65-tk into mouse FM3Atk- cells. The copy numbers of p65-tk in FM3Atk+ cells were 100 to 200 copies per cell. Furthermore, the p65-tk replicated semiconservatively, and the initiation of DNA replication started from the mouse DNA sequences when the replicating activity of p65-tk was tested in the in vitro DNA replication system developed from the FM3A cells. These results show that a 2.5-kilobase fragment of mouse DNA contains the autonomously replicating sequences. PMID- 3031449 TI - Two related regulatory sequences are required for maximal induction of Saccharomyces cerevisiae his3 transcription. AB - In Saccharomyces cerevisiae, the coordinate induction of his3 and other amino acid biosynthesis genes is mediated by the binding of GCN4 activator protein to specific promoter sequences. The his3 regulatory region contains the sequence TGACTC, which with some variation is repeated six times upstream of the mRNA initiation site. The requirements for maximal his3 induction were examined with a series of sequential 5' deletion mutations as well as a set of small internal deletions. Deletions encroaching as far downstream as position -142 behave indistinguishably from the wild-type gene, thus indicating that the two proximal copies of the regulatory sequence are sufficient for maximal induction. Deletions with breakpoints between -137 and -99 confer inducibility, but not to the normal wild-type level. A deletion ending immediately upstream of the proximal TGACTC sequence (position -99) shows some constitutive expression that is independent of the gcn4 gene product. Deletions extending to -94 or beyond do not produce detectable levels of his3 mRNA. Small internal deletions that only remove the proximal regulatory sequence and a 1-base-pair deletion of the thymine residue at -99 abolish induction, but do not affect the basal level of transcription. These results indicate that the proximal copy between -99 and -94 is absolutely required for his3 induction, whereas the copy between -142 and -137 is required only for the maximal level of induction and is inactive by itself. From these and other observations, we suggest the possibility that these related regulatory sequences may be targets for two distinct proteins. PMID- 3031450 TI - Pre-mRNA splicing and the nuclear matrix. AB - We examined the relationship between pre-mRNA splicing and the nuclear matrix by using an in vivo system that we have developed. Plasmids containing the inducible herpesvirus tk gene promoter linked to an intron-containing segment of the rabbit beta-globin gene were transfected into HeLa cells, and then the promoter was transactivated by infection with a TK- virus. Northern analysis revealed that the globin pre-mRNA and all its splicing intermediates and products are associated with the nuclear matrix prepared from such transfected cells. When the nuclear matrix was incubated with a HeLa cell in vitro splicing extract in the presence of ATP, the amount of matrix-associated precursor progressively decreased without a temporal lag in the reaction, with a corresponding increase in free intron lariat. Thus, most of the events of the splicing process (endonucleolytic cuts and branching) occur in this in vitro complementation reaction. However, ligation of exons cannot be monitored in this system because of the abundance of preexisting mature mRNA. Since the matrix is not a self-splicing entity, whereas the in vitro splicing system cannot process efficiently deproteinized matrix RNA, we conclude from our in vitro complementation results (which can be reproduced by using micrococcal nuclease-treated splicing extract) that the nuclear matrix preparation retains parts of preassembled ribonucleoprotein complexes that have the potential to function when supplemented with soluble factors (presumably other than most of the small nuclear ribonucleoproteins known to participate in splicing) present in the HeLa cell extract. PMID- 3031451 TI - Signal peptide specificity in posttranslational processing of the plant protein phaseolin in Saccharomyces cerevisiae. AB - We linked the cDNA coding region for the bean storage protein phaseolin to the promoter and regulatory region of the Saccharomyces cerevisiae repressible acid phosphatase gene (PHO5) in multicopy expression plasmids. Yeast transformants containing these plasmids expressed phaseolin at levels up to 3% of the total soluble cellular protein. Phaseolin polypeptides in S. cerevisiae were glycosylated, and their molecular weights suggested that the signal peptide had been processed. We also constructed a series of plasmids in which the phaseolin signal-peptide-coding region was either removed or replaced with increasing amounts of the amino-terminal coding region for acid phosphatase. Phaseolin polypeptides with no signal peptide were not posttranslationally modified in S. cerevisiae. Partial or complete substitution of the phaseolin signal peptide with that from acid phosphatase dramatically inhibited both signal peptide processing and glycosylation, suggesting that some specific feature of the phaseolin signal amino acid sequence was required for these modifications to occur. Larger hybrid proteins that included approximately one-half of the acid phosphatase sequence linked to the amino terminus of the mature phaseolin polypeptide did undergo proteolytic processing and glycosylation. However, these polypeptides were cleaved at several sites that are not normally used in the unaltered acid phosphatase protein. PMID- 3031452 TI - Localization of specific topoisomerase I interactions within the transcribed region of active heat shock genes by using the inhibitor camptothecin. AB - Camptothecin stabilizes the topoisomerase I-DNA covalent intermediate that forms during the relaxation of torsionally strained DNA. By mapping the position of the resultant DNA nicks, we analyzed the distribution of the covalent intermediates formed on heat shock genes in cultured Drosophila melanogaster cells. Topoisomerase I was found to interact with the transcriptionally active genes hsp22, hsp23, hsp26, and hsp28 after heat shock but not with the inactive genes before heat shock. The interaction occurred predominantly within the transcribed region, with specific sites occurring on both the transcribed and nontranscribed strands of the DNA. Little interaction was seen with nontranscribed flanking sequences. Camptothecin only partially inhibited transcription of the hsp28 gene during heat shock, causing a reduced level of transcripts which were nonetheless full length. Topoisomerase I also interacted with the DNA throughout the transcriptionally active hsp83 gene, including an intron, in both heat-shocked and non-heat-shocked cells. The results point to a dynamic set of interactions at the active locus. PMID- 3031453 TI - Identification of the sequences controlling cyclic AMP regulation and cell-type specific expression of a prestalk-specific gene in Dictyostelium discoideum. AB - We have cloned and analyzed a developmentally and spatially regulated prestalk cell-specific gene from Dictyostelium discoideum. The gene encodes a protein highly homologous to the lysosomal cysteine proteinases cathepsin H and cathepsin B. Amino acid comparisons between these enzymes showed that the active-site amino acids were conserved, as were amino acids known to be important for catalysis and residues which form the intramolecular cysteine bridges. We have constructed a series of internal deletions, duplications, and linker scanner mutations within the region 300 base pairs 5' to the cap site. Analysis of expression of the mutations in transformants identified a approximately 35-base pair GC-rich region containing a dAdC/dGdT palindromic repeat and a G-rich box which is homologous to the 3' GT half of the palindromic repeat. Deletion or disruption of the G box resulted in a approximately 50-fold drop in the level of expression of the gene fusion in transformants in response to cyclic AMP in single-cell culture but did not affect the temporal pattern of regulation or control by cyclic AMP. The expression of such constructs during normal multicellular differentiation paralleled that of the endogenous gene; however, the level of RNA from the constructs was only approximately 10-fold lower than that of constructs containing the G box. Deletion of the 3' half of the palindromic sequence and the G box region resulted in a dramatic decrease in the level of transcription, although the constructs still showed proper temporal expression. These results suggest that this 35-base-pair region acts as an important part of the regulatory region for cell type and cyclic AMP regulation. PMID- 3031454 TI - Human immunoglobulin kappa gene enhancer: chromatin structure analysis at high resolution. AB - The murine immunoglobulin kappa gene enhancer has previously been found to coincide with a region of altered chromatin structure reflected in a DNase I hypersensitivity site detectable on Southern blots of B-cell DNA. We examined the chromatin structure of the homologous region of human DNA using the high resolution electroblotting method originally developed for genomic sequence analysis by G. Church and W. Gilbert (Proc. Natl. Acad. Sci. USA 81:1991-1995, 1984). Analysis of DNA isolated from cells treated in vivo with dimethyl sulfate revealed two B-cell-specific sites of enhanced guanine methylation. Both sites are located within perfect inverted repeats theoretically capable of forming cruciform structures; one of these repeats overlaps an enhancer core sequence. No enhancement or protection of guanine methylation was observed within sequences similar to sites of altered methylation previously described in the immunoglobulin heavy-chain enhancer. Treatment of isolated nuclei with DNase I or a variety of restriction endonucleases defined a B-cell-specific approximately 0.25-kilobase region of enhanced nuclease susceptibility similar to that observed in the murine kappa enhancer. The 130-base-pair DNA segment that shows high sequence conservation between human, mouse, and rabbit DNAs lies at the 5' end of the nuclease-susceptible region. PMID- 3031455 TI - The membrane-associated enzyme phosphatidylserine synthase is regulated at the level of mRNA abundance. AB - To precisely define the functional sequence of the CHO1 gene from Saccharomyces cerevisiae, encoding the regulated membrane-associated enzyme phosphatidylserine synthase (PSS), we subcloned the original 4.5-kilobase (kb) CHO1 clone. In this report a 2.8-kb subclone was shown to complement the ethanolamine-choline auxotrophy and to repair the defect in the synthesis of phosphatidylserine, both of which are characteristic of cho1 mutants. When this subclone was used as a hybridization probe of Northern and slot blots of RNA from wild-type cells, the abundance of a 1.2-kb RNA changed in response to alterations in the levels of the soluble phospholipid precursors inositol and choline. The addition of inositol led to a 40% repression of the 1.2-kb RNA level, while the addition of choline and inositol led to an 85% repression. Choline alone had little repressive effect. The level of 1.2-kb RNA closely paralleled the level of PSS activity found in the same cells as determined by enzyme assays. Disruption of the CHO1 gene resulted in the simultaneous disappearance of 1.2-kb RNA and PSS activity. Cells bearing the ino2 or ino4 regulatory mutations, which exhibit constitutively repressed levels of a number of phospholipid biosynthetic enzymes, had constitutively repressed levels of 1.2-kb RNA and PSS activity. Another regulatory mutation, opi1, which causes the constitutive derepression of PSS and other phospholipid biosynthetic enzymes, caused the constitutive derepression of the 1.2-kb RNA. When cho1 mutant cells were transformed with the 2.8-kb subclone on a single-copy plasmid, the 1.2-kb RNA and PSS activity levels were regulated in a wild-type fashion. The presence of the 2.8-kb subclone on a multicopy plasmid, however, led to the constitutive overproduction of 1.2-kb RNA and PSS in cho1 cells. PMID- 3031456 TI - Isolation and characterization of MOD5, a gene required for isopentenylation of cytoplasmic and mitochondrial tRNAs of Saccharomyces cerevisiae. AB - The mod5-1 mutation is a nuclear mutation in Saccharomyces cerevisiae that reduces the biosynthesis of N6-(delta 2-isopentenyl)adenosine in both cytoplasmic and mitochondrial tRNAs to less than 1.5% of wild-type levels. The tRNA modification enzyme, delta 2-isopentenyl pyrophosphate:tRNA isopentenyl transferase, cannot be detected in vitro with extracts from mod5-1 cells. A characterization of the MOD5 gene would help to determine how the same enzyme activity in different cellular compartments can be abolished by a single nuclear mutation. To that end we have cloned the MOD5 gene and shown that it restores delta 2-isopentenyl pyrophosphate:tRNA isopentenyl transferase activity and N6 (delta 2-isopentenyl)adenosine to tRNA in both the mitochondria and the nucleus/cytoplasm compartments of mod5-1 yeast cells. That MOD5 sequences are expressed in Escherichia coli and can complement an N6-(delta 2-isopentenyl)-2 methylthioadenosine-deficient E. coli mutant leads us to conclude that MOD5 is the structural gene for delta 2-isopentenyl pyrophosphate:tRNA isopentenyl transferase. PMID- 3031442 TI - Antimicrobial resistance of Staphylococcus aureus: genetic basis. PMID- 3031458 TI - Bidirectional activity of the rat insulin II 5'-flanking region in transgenic mice. AB - We have identified a new transcription initiation site in the 5'-flanking regulatory region of the rat insulin II gene. This site is located on the opposite strand with respect to the insulin gene promoter, upstream of the insulin gene transcriptional enhancer. The cell-specific activity of this reverse promoter element is demonstrated in two lineages of transgenic mice, in which it directs expression of simian virus 40 T antigen specifically to the beta cells of the endocrine pancreas, resulting in development of pancreatic tumors. Analysis of RNA from the tumor cells demonstrates bidirectional transcription from the insulin regulatory region of the transgene. These data raise the possibility that bidirectional activity is a quality of the regulatory region of the insulin gene in its natural genomic context. PMID- 3031457 TI - DNA sequence and transcript mapping of MOD5: features of the 5' region which suggest two translational starts. AB - A mutation in the yeast nuclear gene MOD5 drastically reduces the biosynthesis of the modified base isopentenyladenosine in tRNAs located in different cellular compartments: the mitochondria and the nucleus or cytoplasm. Several lines of evidence strongly suggest that MOD5 is the structural gene encoding the tRNA modifying enzyme delta 2-isopentenyl pyrophosphate:tRNA isopentenyl transferase. DNA sequence analysis of MOD5 reveals an open reading frame of 428 amino acids. A set of mRNAs heterogeneous at both the 5' and 3' termini are transcribed from this gene. Although all of these transcripts initiate upstream of the first AUG codon of the open reading frame, a subset has an extremely short (greater than or equal to 1 base) 5' leader. Furthermore, in positions important for efficient initiation of translation and generally occupied by purines, this first AUG codon is flanked by a U (position -3) and a C (position +4). It is possible that two proteins, one with an amino-terminal extension of basic charge, could be generated from the MOD5 gene via differential translational starts. PMID- 3031459 TI - Directed mutagenesis in Candida albicans: one-step gene disruption to isolate ura3 mutants. AB - A method for introducing specific mutations into the diploid Candida albicans by one-step gene disruption and subsequent UV-induced recombination was developed. The cloned C. albicans URA3 gene was disrupted with the C. albicans ADE2 gene, and the linearized DNA was used for transformation of two ade2 mutants, SGY-129 and A81-Pu. Both an insertional inactivation of the URA3 gene and a disruption which results in a 4.0-kilobase deletion were made. Southern hybridization analyses demonstrated that the URA3 gene was disrupted on one of the chromosomal homologs in 15 of the 18 transformants analyzed. These analyses also revealed restriction site dimorphism of EcoRI at the URA3 locus which provides a unique marker to distinguish between chromosomal homologs. This enabled us to show that either homolog could be disrupted and that disrupted transformants of SGY-129 contained more than two copies of the URA3 locus. The A81-Pu transformants heterozygous for the ura3 mutations were rendered homozygous and Ura- by UV induced recombination. The homozygosity of a deletion mutant and an insertion mutant was confirmed by Southern hybridization. Both mutants were transformed to Ura+ with plasmids containing the URA3 gene and in addition, were resistant to 5 fluoro-orotic acid, a characteristic of Saccharomyces cerevisiae ura3 mutants as well as of orotidine-5'-phosphate decarboxylase mutants of other organisms. PMID- 3031460 TI - Development of autonomously replicating plasmids for Candida albicans. AB - A pool of Candida albicans RsaI fragments cloned onto a vector containing pBR322 sequences and the Candida ADE2 gene was used to transform a Candida ade2 mutant to adenine protrophy. A potential autonomously replicating sequence (ARS) in Candida DNA was identified by two criteria: instability of the selectable marker in the absence of selection and the presence of free plasmid in total DNA preparations. Plasmids carrying the ARS transformed C. albicans at a high frequency (200 to 1,000 ADE+ transformants per microgram of DNA), and Southern hybridization analysis of these transformants indicated that multiple copies of the plasmid sequences were present and that, although they were present in high molecular-weight molecules, these sequences had not undergone rearrangement. Orthogonal field alternation gel electrophoresis indicated that the high molecular-weight transforming sequences were not associated with any chromosome. The simplest interpretation to account for these data is that the transforming sequences are present as oligomers consisting of head-to-tail tandem repeats. The transformed strains occasionally yield stable segregants in which the transforming sequences are integrated into the chromosome as repeats. The Candida sequence responsible for the ARS phenotype was limited to a single 0.35-kilobase RsaI fragment which is present in one copy per haploid genome. PMID- 3031461 TI - DNA mismatch repair detected in human cell extracts. AB - A system to study mismatch repair in vitro in HeLa cell extracts was developed. Preformed heteroduplex plasmid DNA containing two single base pair mismatches within the SupF gene of Escherichia coli was used as a substrate in a mismatch repair assay. Repair of one or both of the mismatches to the wild-type sequence was measured by transformation of a lac(Am) E. coli strain in which the presence of an active supF gene could be scored. The E. coli strain used was constructed to carry mutations in genes associated with mismatch repair and recombination (mutH, mutU, and recA) so that the processing of the heteroduplex DNA by the bacterium was minimal. Extract reactions were carried out by the incubation of the heteroduplex plasmid DNA in the HeLa cell extracts to which ATP, creatine phosphate, creatine kinase, deoxynucleotides, and a magnesium-containing buffer were added. Under these conditions about 1% of the mismatches were repaired. In the absence of added energy sources or deoxynucleotides, the activity in the extracts was significantly reduced. The addition of either aphidicolin or dideoxynucleotides reduced the mismatch repair activity, but only aphidicolin was effective in blocking DNA polymerization in the extracts. It is concluded that mismatch repair in these extracts is an energy-requiring process that is dependent on an adequate deoxynucleotide concentration. The results also indicate that the process is associated with some type of DNA polymerization, but the different effects of aphidicolin and dideoxynucleotides suggest that the mismatch repair activity in the extracts cannot simply be accounted for by random nick translation activity alone. PMID- 3031462 TI - Nucleotide sequence analysis of a cloned DNA fragment from human cells reveals homology to retrotransposons. AB - During molecular cloning of proviral DNA of human spumaretrovirus, various recombinant clones were established and analyzed. Blot hybridization revealed that one of the recombinant plasmids had the characteristic features of a member of the long interspersed repetitive sequences family. The DNA element was analyzed by restriction mapping and nucleotide sequencing. It showed a high degree of amino acid sequence homology of 54.3% when compared with the 5' terminal part of the pol gene product of the murine retrotransposon LIMd. The 3' region of the cloned DNA element encodes proteins with an even higher degree of homology of 67.4% in comparison to the corresponding parts of a member of the primate KpnI sequence family. PMID- 3031463 TI - Heat shock response of Saccharomyces cerevisiae mutants altered in cyclic AMP dependent protein phosphorylation. AB - When Saccharomyces cerevisiae cells grown at 23 degrees C were transferred to 36 degrees C, they initiated synthesis of heat shock proteins, acquired thermotolerance to a lethal heat treatment given after the temperature shift, and arrested their growth transiently at the G1 phase of the cell division cycle. The bcy1 mutant which resulted in production of cyclic AMP (cAMP)-independent protein kinase did not synthesize the three heat shock proteins hsp72A, hsp72B, and hsp41 after the temperature shift. The bcy1 cells failed to acquire thermotolerance to the lethal heat treatment and were not arrested at the G1 phase after the temperature shift. In contrast, the cyr1-2 mutant, which produced a low level of cAMP, constitutively produced three heat shock proteins and four other proteins without the temperature shift and was resistant to the lethal heat treatment. The results suggest that a decrease in the level of cAMP-dependent protein phosphorylation results in the heat shock response, including elevated synthesis of three heat shock proteins, acquisition of thermotolerance, and transient arrest of the cell cycle. PMID- 3031465 TI - A highly conserved endonuclease activity present in Escherichia coli, bovine, and human cells recognizes oxidative DNA damage at sites of pyrimidines. AB - We have compared the sites of nucleotide incision on DNA damaged by oxidizing agents when cleavage is mediated by either Escherichia coli endonuclease III or an endonuclease present in bovine and human cells. E. coli endonuclease III, the bovine endonuclease isolated from calf thymus, and the human endonuclease partially purified from HeLa and CEM-C1 lymphoblastoid cells incised DNA damaged with osmium tetroxide, ionizing radiation, or high doses of UV light at sites of pyrimidines. For each damaging agent studied, regardless of whether the E. coli, bovine, or human endonuclease was used, the same sequence specificity of cleavage was observed. We detected this endonuclease activity in a variety of human fibroblasts derived from normal individuals as well as individuals with the DNA repair deficiency diseases ataxia telangiectasia and xeroderma pigmentosum. The highly conserved nature of such a DNA damage-specific endonuclease suggests that a common pathway exists in bacteria, humans, and other mammals for the reversal of certain types of oxidative DNA damage. PMID- 3031464 TI - Ty1 sequence with enhancer and mating-type-dependent regulatory activities. AB - Some insertion mutations in Saccharomyces cerevisiae activate the expression of adjacent structural genes. The CYC7-H2 mutation is a Ty1 insertion 5' to the iso 2-cytochrome c coding region of CYC7. The Ty1 insertion causes a 20-fold increase in CYC7 expression in a and alpha haploid cell types of S. cerevisiae. This activation is repressed in the a/alpha diploid cell type. Previous computer analysis of the CYC7-H2 Ty1 activator region identified two related sequences with homology both to mammalian enhancers and to a yeast a/alpha control site. A 112-base-pair (bp) DNA fragment encompassing one of these blocks of homology functioned as one component of the Ty1 activator. A 28-bp synthetic oligonucleotide with the wild-type homology block sequence was also functional. A single base pair mutation within the enhancer core of the synthetic 28-bp regulatory element reduced its activation ability to near background amounts. In addition, the 112-bp Ty1 fragment by itself functioned as a target for repression of adjacent gene expression in a/alpha diploid cells. PMID- 3031466 TI - Transcription of spacer sequences flanking the rat 45S ribosomal DNA gene. AB - The transcriptional activity of spacer sequences flanking the rat 45S ribosomal DNA (rDNA) gene were studied. Nascent RNA labeled in in vitro nuclear run-on reactions hybridized with both 5' and 3' spacer regions. The highest level of hybridization was seen with an rDNA fragment containing tandem repeats of a 130 base-pair sequence upstream of the 45S rRNA initiation site. Synthesis of RNA transcripts homologous to this internally repetitious spacer region was insensitive to high levels of alpha-amanitin, suggesting that it is mediated by RNA polymerase I. Analysis of steady-state RNA showed that these transcripts were present at extremely low levels in vivo relative to precursor rRNA transcripts. In contrast, precursor and spacer run-on RNAs were synthesized at similar levels. This suggests that spacer transcripts are highly unstable in vivo; therefore, it may be the process of transcription rather than the presence of spacer transcripts that is functionally important. Transcription in this upstream rDNA region may be involved in regulation of 45S rRNA synthesis in rodents, as has been suggested previously for frog rRNA. In addition, the presence of transcriptional activity in other regions of the spacer suggests that some polymerase I molecules may transcribe through the spacer from one 45S gene to the next on rodent rDNA. PMID- 3031467 TI - Frequent independent duplicative transpositions activate a single VSG gene. AB - The expression of several surface antigen genes in Trypanosoma brucei is mediated by the duplicative transposition of a basic-copy variant surface glycoprotein (VSG) gene into an expression site. We determined that the appearance of variant 118, in a parasitemia, resulted from at least four independent duplicative transpositions of the same VSG 118 gene. Variants 117 and 118 both appeared at specific periods but resulted from multiple independent activations. Antigenic variants thus occur in an ordered manner. We show that in the duplicative transpositions of VSG genes, the ends of the transposed segments were homologous between the basic copy and the expression site. Sequences other than the previously reported 70-base-pair (bp) repeats could be involved. In one variant, 118 clone 1, the homology was between a sequence previously transposed into the expression site and a sequence located 6 kilobases upstream of the VSG 118 gene. In variant 118b the homology was presumably in 70-bp repeat arrays, while in a third 118 variant yet another sequence was involved. The possibility that the 70 bp repeats are important in the initial steps of the recombinational events was illustrated by a rearrangement involving a 70-bp repeat array. The data provide strong evidence for the notion that gene conversion mediates the duplicative transposition of VSG genes. We discuss a model that explains how the process of duplicative transposition can occur at random and still produce an ordered appearance of variants. PMID- 3031468 TI - Phosphorylation of talin at tyrosine in Rous sarcoma virus-transformed cells. AB - The cytoskeletal protein talin was found to undergo enhanced phosphorylation at tyrosine residues in chicken embryo fibroblasts following transformation by Rous sarcoma virus. An increase in the tyrosine phosphorylation of talin was also observed within 6 h in cells infected by the temperature-sensitive mutant tsNY68 after a shift from the nonpermissive to the permissive temperature. The overall extent of phosphorylation was 0.07 mol of phosphate per mol of talin and was not appreciably altered by transformation. In uninfected cells talin was shown to be phosphorylated at multiple sites by tryptic peptide mapping. Following transformation most of these sites remained phosphorylated, to the same or to a lesser extent, while novel, phosphotyrosine-containing phosphopeptides appeared. Talin was phosphorylated at tyrosine in cells infected by Rous sarcoma virus mutants which induce altered or partial transformation morphologies; thus the increased phosphorylation of talin at tyrosine occurred irrespective of the morphology induced. Transformation by Y73 also induced elevated levels of phosphotyrosine in talin, whereas transformation by the avian erythroblastosis and Fujinami sarcoma viruses did not. PMID- 3031469 TI - Analysis of mutation in human cells by using an Epstein-Barr virus shuttle system. AB - We developed highly sensitive shuttle vector systems for detection of mutations formed in human cells using autonomously replicating derivatives of Epstein-Barr virus (EBV). EBV vectors carrying the bacterial lacI gene as the target for mutation were established in human cells and later returned to Escherichia coli for rapid detection and analysis of lacI mutations. The majority of the clonal cell lines created by establishment of the lacI-EBV vector show spontaneous LacI- frequencies of less than 10(-5) and are suitable for studies of induced mutation. The ability to isolate clonal lines represents a major advantage of the EBV vectors over transiently replicating shuttle vectors (such as those derived from simian virus 40) for the study of mutation. The DNA sequence changes were determined for 61 lacI mutations induced by exposure of one of the cell lines to N-nitroso-N-methylurea. A total of 33 of 34 lacI nonsense mutations and 26 of 27 missense mutations involve G X C to A X T transitions. These data provide support for the mutational theory of cancer. PMID- 3031470 TI - cis-acting regulatory elements within gag genes of avian retroviruses. AB - A cis-acting enhancer element has been detected within the gag gene of several avian retroviruses, including Rous sarcoma virus, Fujinami sarcoma virus, and the endogenous Rous-associated virus-0. A consensus enhancer core sequence, GTGGTTTG, is present in all of these viral genomes, approximately 900 bases downstream from the site of initiation of transcription. When an internal fragment derived from the gag gene of any of these viruses (spanning nucleotides 533 to approximately 1149) was inserted into a plasmid containing the chloramphenicol acetyltransferase (cat) gene under control of the simian virus 40 promoter, 9- or 21-fold enhancement of CAT expression was observed after transfection into mouse L cells and chicken embryo fibroblasts, respectively. This enhancement was not dependent on the position of insertion of the gag fragment into the plasmid. However, there was a strong dependence on orientation, with higher levels of CAT expression in constructs in which the 5' end of the gag fragment was nearest to the promoter, suggesting a possible negative regulatory element at the 3' end of this fragment. Deletion of the 3' end of the insert resulted in a gag fragment, containing nucleotides 533 to 1017, which enhanced expression equally in either orientation. When the gag fragment was inserted into a plasmid containing the cat gene under the control of an intact Rous sarcoma virus long terminal repeat, it induced a two- to threefold increase in CAT activity and CAT mRNA levels. Translation of the gag fragment did not appear to be necessary for the observed enhancement, since two insertional mutations resulting in frameshifts in the gag insert did not affect CAT expression. However, deletion of a 330-base internal fragment from the gag insert restored a basal level of CAT activity. These results suggest that retroviruses have regulatory elements within their genes distinct from those in the long terminal repeats that flank the genes. PMID- 3031471 TI - Molecular cloning of chromosome I DNA from Saccharomyces cerevisiae: isolation and analysis of the CEN1-ADE1-CDC15 region. AB - To continue the systematic examination of the physical and genetic organization of an entire Saccharomyces cerevisiae chromosome, the DNA from the CEN1-ADE1 CDC15 region from chromosome I was isolated and characterized. Starting with the previously cloned ADE1 gene (J. C. Crowley and D. B. Kaback, J. Bacteriol. 159:413-417, 1984), a series of recombinant lambda bacteriophages containing 82 kilobases of contiguous DNA from chromosome I were obtained by overlap hybridization. The cloned sequences were mapped with restriction endonucleases and oriented with respect to the genetic map by determining the physical positions of the CDC15 gene and the centromeric DNA (CEN1). The CDC15 gene was located by isolating plasmids from a YCp50 S. cerevisiae genomic library that complemented the cdc15-1 mutation. S. cerevisiae sequences from these plasmids were found to be represented among those already obtained by overlap hybridization. The cdc15-1-complementing plasmids all shared only one intact transcribed region that was shown to contain the bona fide CDC15 gene by in vitro gene disruption and one-step replacement to delete the chromosomal copy of this gene. This deletion produced a recessive lethal phenotype that was also recessive to cdc15-1. CEN1 was located by finding a sequence from the appropriate part of the cloned region that stabilized the inheritance of autonomously replicating S. cerevisiae plasmid vectors. Finally, RNA blot hybridization and electron microscopy of R-loop-containing DNA were used to map transcribed regions in the 23 kilobases of DNA that went from CEN1 to CDC15. In addition to the transcribed regions corresponding to the ADE1 and ADC15 genes, this DNA contained five regions that gave rise to polyadenylated RNA, at least two regions complementary to 4S RNA species, and a Ty1 transposable element. Notably, a higher than average proportion of the DNA examined was transcribed into RNA. PMID- 3031472 TI - Nucleotide sequence structure and consistency of a developmentally regulated DNA deletion in Tetrahymena thermophila. AB - DNA deletion by site-specific chromosome breakage and rejoining occurs extensively during macronuclear development in the ciliate Tetrahymena thermophila. We have sequenced both the micronuclear (germ line) and rearranged macronuclear (somatic) forms of one region from which 1.1 kilobases of micronuclear DNA are reproducibly deleted during macronuclear development. The deletion junctions lie within a pair of 6-base-pair direct repeats. The termini of the deleted sequence are not inverted repeats. The precision of deletion at the nucleotide level was also characterized by hybridization with a synthetic oligonucleotide matching the determined macronuclear (rejoined) junction sequence. This deletion occurs in a remarkably sequence-specific manner. However, a very minor degree of variability in the macronuclear junction sequences was detected and was shown to be inherent in the mechanism of deletion itself. These results suggest that DNA deletion during macronuclear development in T. thermophila may constitute a novel type of DNA recombination and that it can create sequence heterogeneity on the order of a few base pairs at rejoining junctions. PMID- 3031473 TI - The viral Ki-ras gene must be expressed in the G2 phase if ts Kirsten sarcoma virus-infected NRK cells are to proliferate in serum-free medium. AB - NRK cells infected with a temperature-sensitive Kirsten sarcoma virus (ts371 KSV) are transformed at 36 degrees C, but are untransformed at 41 degrees C which inactivates the abnormally thermolabile oncogenic p21Ki product of the viral Ki ras gene. At 41 degrees C, tsKSV-infected NRK cells were arrested in G0/G1 when incubated in serum-free medium, but could then be stimulated to transit G1, replicate DNA, and divide by adding serum at 41 degrees C or dropping the temperature to a p21-activating 36 degrees C without adding serum. When quiescent cells at 41 degrees C were stimulated to transit G1 in serum-free medium by activating p21 at 36 degrees C and then shifted back to the p21-inactivating 41 degrees C in the mid-S phase, they continued replicating DNA but could not transit G2. Reactivating p21 in the G2-arrested cells by once again lowering the temperature to 36 degrees C stimulated a rapid entry into mitosis. By contrast, while serum-stimulated quiescent G0 cells at 41 degrees C replicate DNA and divide, serum did not induce G2-arrested cells to enter mitosis, indicating that serum growth factors may trigger events in the G1 phase that ultimately determine G2 transit. These observations made with the viral ras product suggest that cellular ras proto-oncogene products have a role in G2 transit of normal cells. PMID- 3031474 TI - Rate of replication of the murine immunoglobulin heavy-chain locus: evidence that the region is part of a single replicon. AB - We measured the temporal order of replication of EcoRI segments from the murine immunoglobulin heavy-chain constant region (IgCH) gene cluster, including the joining (J) and diversity (D) loci and encompassing approximately 300 kilobases. The relative concentrations of EcoRI segments in bromouracil-labeled DNA that replicated during selected intervals of the S phase in Friend virus-transformed murine erythroleukemia (MEL) cells were measured. From these results, we calculated the nuclear DNA content (C value; the haploid DNA content of a cell in the G1 phase of the cell cycle) at the time each segment replicated during the S phase. We observed that IgCH genes replicate in the following order: alpha, epsilon, gamma 2a, gamma 2b, gamma 1, gamma 3, delta, and mu, followed by the J and D segments. The C value at which each segment replicates increased as a linear function of its distance from C alpha. The average rate of DNA replication in the IgCH gene cluster was determined from these data to be 1.7 to 1.9 kilobases/min, similar to the rate measured for mammalian replicons by autoradiography and electron microscopy (for a review, see H. J. Edenberg and J. A. Huberman, Annu. Rev. Genet. 9:245-284, 1975, and R. G. Martin, Adv. Cancer Res. 34:1-55, 1981). Similar results were obtained with other murine non-B cell lines, including a fibroblast cell line (L60T) and a hepatoma cell line (Hepa 1.6). In contrast, we observed that IgCh segments in a B-cell plasmacytoma (MPC11) and two Abelson murine leukemia virus-transformed pre-B cell lines (22D6 and 300-19O) replicated as early as (300-19P) or earlier than (MPC11 and 22D6) C alpha in MEL cells. Unlike MEL cells, however, all of the IgCH segments in a given B cell line replicated at very similar times during the S phase, so that a temporal directionality in the replication of the IgCH gene cluster was not apparent from these data. These results provide evidence that in murine non-B cells the IgCH, J, and D loci are part of a single replicon. PMID- 3031475 TI - Cyclic AMP regulation of early gene expression in Dictyostelium discoideum: mediation via the cell surface cyclic AMP receptor. AB - We examined two sets of genes expressed early in the developmental cycle of Dictyostelium discoideum that appear to be regulated by cyclic AMP (cAMP). The transcripts of both sets of genes were not detectable in vegetative cells. During normal development on filter pads, RNA complementary to these genes could be detected at about 2 h, peaked around 6 to 8 h, and decreased gradually thereafter. Expression of these genes upon starvation in shaking culture was stimulated by pulsing the cells with nanomolar levels of cAMP, a condition that mimics the in vivo pulsing during normal aggregation. Expression was inhibited by caffeine or by continuous levels of cAMP, a condition found later in development when in vivo expression of these genes decreased. The inhibition of caffeine could be overcome by pulsing cells with cAMP. These results suggest that the expression is mediated via the cell surface cAMP receptor, but does not require a rise in intracellular cAMP. mRNA from a gene of the second class was induced upon starvation, peaked by 2.5 h of development, and then declined. In contrast to the other genes, its expression was maintained by continuous levels of cAMP and repressed by cAMP pulses. These and other results on a number of classes of developmentally regulated genes indicates that changing levels of cAMP, acting via the cell surface cAMP receptor, are involved in controlling these groups of genes. We also examined the structure and partial sequence of the cAMP pulse induced genes. The two tandemly duplicated M3 genes were almost continuously homologous over the sequenced portion of the protein-coding region except for a region near the N-terminal end. The two M3 genes had regions of homology in the 5' flanking sequence and showed slight homology to the same regions in gene D2, another cAMP pulse-induced gene. D2 showed extremely significant homology over its entire sequenced length to an acetylcholinesterase. The results presented here and by others suggest that expression of many early genes in D. discoideum is regulated via the cell surface cAMP receptor. We expect that many of these genes may play essential roles in early Dictyostelium development and could code for elements of the cAMP signal transduction pathway involved in aggregation. PMID- 3031476 TI - Isolation, characterization, and expression of the gene encoding the late histone subtype H1-gamma of the sea urchin Strongylocentrotus purpuratus. AB - We cloned and characterized the gene encoding H1-gamma, a late histone subtype of the sea urchin species Strongylocentrotus purpuratus. The predicted primary sequence of H1-gamma is 216 amino acids in length and has a net charge of +70, which is high for a somatic H1 histone. The H1-gamma gene appears to be a unique sequence gene that is not tightly linked to the core histone genes. The 770-base pair transcribed region of the H1-gamma gene is bordered on the 5' side by two previously described H1-specific sequence elements and on the 3' side by a hairpin loop structure and CAGA box sequences. We detected 3,900 stored maternal H1-gamma mRNA transcripts per egg. The number of H1-gamma transcripts per embryo rises by 9.5 h postfertilization, but the maximum rate of accumulation (4,300 molecules per min per embryo) occurs in the late-blastula-stage embryo between 14 and 21 h after fertilization. The number of H1-gamma mRNA molecules peaks 21 h after fertilization when there are 2.0 X 10(6) molecules per embryo (a 500-fold increase) and then decreases over the next 3.25 h to 1.3 million molecules per embryo. Between 24 and 82 h after fertilization the number of H1-gamma transcripts declines steadily (210 molecules per min per embryo) to reach approximately 5.4 X 10(5) H1-gamma mRNAs by 82 h postfertilization. Surprisingly, the number of late H1 mRNA molecules per embryo is greater than the number of late H2B mRNA molecules beginning at the early gastrula stage of development. PMID- 3031477 TI - Requirements for accurate and efficient mRNA 3' end cleavage and polyadenylation of a simian virus 40 early pre-RNA in vitro. AB - Using a pre-RNA containing the simian virus 40 early introns and poly(A) addition site, we investigated several possible requirements for accurate and efficient mRNA 3' end cleavage and polyadenylation in a HeLa cell nuclear extract. Splicing and 3' end formation occurred under the same conditions but did not appear to be coupled in any way in vitro. Like splicing, 3' end cleavage and polyadenylation each required Mg2+, although spermidine could substitute in the cleavage reaction. Additionally, cleavage of this pre-RNA, but not others, was totally blocked by EDTA, indicating that structural features of pre-RNA may affect the ionic requirements of 3' end formation. The ATP analog 3' dATP inhibited both cleavage and polyadenylation even in the presence of ATP, possibly reflecting the coupled nature of these activities. A 5' cap structure appears not to be required for mRNA 3' end processing in vitro because neither the presence or absence of a 5' cap on the pre-RNA nor the addition of cap analogs to reaction mixtures had any effect on the efficiency of 3' end processing. Micrococcal nuclease pretreatment of the nuclear extract inhibited cleavage and polyadenylation. However, restoration of activity was achieved by addition of purified Escherichia coli RNA, suggesting that the inhibition caused by such a nuclease treatment was due to a general requirement for mass of RNA rather than to destruction of a particular nucleic acid-containing component such as a small nuclear ribonucleoprotein. PMID- 3031478 TI - Sucl+ encodes a predicted 13-kilodalton protein that is essential for cell viability and is directly involved in the division cycle of Schizosaccharomyces pombe. AB - Sucl+ was originally identified as a DNA sequence that, at high copy number, rescued Schizosaccharomyces pombe strains carrying certain temperature-sensitive alleles of the cdc2 cell cycle control gene. We determined the nucleotide sequence of a 1,083-base-pair Sucl+ DNA fragment and S1 mapped its 866-nucleotide RNA transcript. The protein-coding sequence of the gene is interrupted by two intervening sequences of 115 and 51 base pairs. The predicted translational product of the gene is a protein of 13 kilodaltons. A chromosomal gene disruption of Sucl+ was constructed in a diploid S. pombe strain. Germinating spores carrying a null allele of the gene were capable of very limited cell division, following which many cells became highly elongated. The Sucl+ gene was also strongly overexpressed under the control of a heterologous S. pombe promoter. Overexpression of Sucl+ is not lethal but causes a division delay such that cells are approximately twice the normal length at division. These data suggest that Sucl+ encodes a protein which plays a direct role in the cell division cycle of S. pombe. PMID- 3031479 TI - Identification of a new common provirus integration site in gross passage A murine leukemia virus-induced mouse thymoma DNA. AB - The Gross passage A murine leukemia virus (MuLV) induced T-cell leukemia of clonal (or oligoclonal) origin in inoculated mice. To study the role of the integrated proviruses in these tumor cells, we cloned several newly integrated proviruses (with their flanking cellular sequences) from a single tumor in procaryotic vectors. With each of the five clones obtained, a probe was prepared from the cellular sequences flanking the provirus. With one such probe (SS8), we screened several Gross passage A MuLV-induced SIM.S mouse tumor DNAs and found that, in 11 of 40 tumors, a provirus was integrated into a common region designated Gin-1. A 26-kilobase-pair sequence of Gin-1 was cloned from two lambda libraries, and a restriction map was derived. All proviruses were integrated as a cluster in the same orientation within a 5-kilobase-pair region of Gin-1, and most of them had a recombinant structure of the mink cell focus-forming virus type. The frequency of Gin-1 occupancy by provirus was much lower in thymoma induced by other strains of MuLV in other mouse strains. Using somatic-cell hybrid DNAs, we mapped Gin-1 on mouse chromosome 19. Gin-1 was not homologous to 16 known oncogenes and was distinct from the other common regions for provirus integration previously described. Therefore, Gin-1 appears to represent a new common provirus integration region. The integration of a provirus within Gin-1 might be an important event leading to T-cell transformation, and the Gin-1 region might harbor sequences which are involved in tumor development. PMID- 3031480 TI - The genome of Shope fibroma virus, a tumorigenic poxvirus, contains a growth factor gene with sequence similarity to those encoding epidermal growth factor and transforming growth factor alpha. AB - Degenerate oligonucleotide probes corresponding to a highly conserved region common to epidermal growth factor, transforming growth factor alpha, and vaccinia growth factor were used to identify a novel growth factor gene in the Shope fibroma virus genome. Sequence analysis indicates that the Shope fibroma growth factor is a distinct new member of this family of growth factors. PMID- 3031481 TI - Induction of c-Ha-ras transcription in rat cells by simian virus 40 large T antigen. AB - Rat 3Y1 cells expressing simian virus 40 large T antigen under the control of the mouse mammary tumor virus long terminal repeat were established. The amount of c Ha-ras mRNA in those cells was elevated by about 20 times in parallel with large T antigen after exposure to dexamethasone for 48 h. In chloramphenicol acetyltransferase assays with a plasmid containing the c-Ha-ras-1 promoter the increase in c-Ha-ras mRNA was shown to occur at the transcriptional level. PMID- 3031482 TI - Functional domains of a T-DNA promoter active in crown gall tumors. AB - Promoter domains required for transcriptional expression of the 780 gene of T right (pTi15955) were identified by deletion mutagenesis. Accurate quantitation of transcriptional activity of a series of 5' and internal deletion mutants was achieved by using a double gene vector containing a reference 780 gene as an internal standard. Results of the 5' deletions delineated an activator element located between -440 and -229 base pairs (bp) from the start of transcription. Removal of this region resulted in a 100-fold decrease in promoter activity. Two relatively small internal deletion/substitution mutations at positions -74 to -76 and -98 to -112 reduced promoter activity to 38 and 42%, respectively. In most cases large-scale internal deletions (38 to 151 bp) occurring in various locations from positions -12 to -348 bp caused a significant loss in major promoter activity. However, three internal deletions starting at position -37 and extending upstream as far as -153 bp either had little effect on transcriptional activity or resulted in increased activity. Removal of the TATA motif drastically reduced promoter activity to less than 0.1% of the wild type. A minor start of transcription was detected 60 bases upstream from the major transcriptional start site. This minor promoter shares the same activator element as the major promoter for full activity. Deletion and insertion mutations downstream of the minor promoter TATA demonstrated the role of the TATA box in positioning the start of transcription. PMID- 3031483 TI - Sedimentation of an RNA polymerase complex from vaccinia virus that specifically initiates and terminates transcription. AB - A high-molecular-weight protein complex that is capable of accurate transcription initiation and termination of vaccinia virus early genes without additional factors was demonstrated. The complex was solubilized by disruption of purified virions, freed of DNA by passage through a DEAE-cellulose column, and isolated by glycerol gradient sedimentation. All detectable RNA polymerase activity was associated with the transcription complex, whereas the majority of enzymes released from virus cores including mRNA (nucleoside-2'-O)methyltransferase, poly(A) polymerase, topoisomerase, nucleoside triphosphate phosphohydrolase II, protein kinase, and single-strand DNase sedimented more slowly. Activities corresponding to two enzymes, mRNA guanylyltransferase (capping enzyme) and nucleoside triphosphate phosphohydrolase I (DNA-dependent ATPase), partially sedimented with the complex. Silver-stained polyacrylamide gels, immunoblots, and autoradiographs confirmed the presence of subunits of vaccinia virus RNA polymerase, mRNA guanylyltransferase, and nucleoside triphosphate phosphohydrolase I, as well as additional unidentified polypeptides, in fractions with transcriptase activity. A possible role for the DNA-dependent ATPase was suggested by studies with ATP analogs with gamma-S or nonhydrolyzable beta-gamma phosphodiester bonds. These analogs were used by vaccinia virus RNA polymerase to nonspecifically transcribe single-stranded DNA templates but did not support accurate transcription of early genes by the complex. Transcription also was sensitive to high concentrations of novobiocin; however, this effect could be attributed to inhibition of RNA polymerase or ATPase activities rather than topoisomerase. PMID- 3031484 TI - Rapid inhibition of processing and assembly of small nuclear ribonucleoproteins after infection with vesicular stomatitis virus. AB - After infection of baby hamster kidney cells with vesicular stomatitis virus (VSV), processing and assembly of small nuclear ribonucleoproteins (snRNP) were rapidly inhibited. The U1 and U2 snRNAs accumulated as precursor species approximately 3 and 10 nucleotides longer, respectively, than the mature RNAs. Alteration in snRNP assembly was noted because the precursor snRNAs were not associated with the U-series RNA-core protein complex in infected cells. However, antibodies specific for the U2 RNA-binding protein, A', were able to precipitate pre-U2 RNAs from VSV-infected cells. These results indicated that precursors to U2 RNA were bound to A' and remained bound during virus infection. Analysis of the synthesis of proteins normally associated with U1 and U2 RNAs indicated that synthesis was unaffected at times when snRNP assembly with core proteins was blocked by the VSV. These findings suggested that the core proteins associate with one another in the absence of the snRNAs in VSV-infected cells. They further suggest a correlation between the inability of the core complex to bind the U series snRNPs and the failure to process the 3' ends of U1 and U2 RNAs in VSV infected cells. These effects of VSV on snRNP assembly may be related to the shutoff of host-cell macromolecular synthesis. PMID- 3031486 TI - Episomal maintenance of a bovine papilloma virus vector in transgenic mice. AB - We have used a bovine papillomavirus-based vector to generate transgenic mice. Transgenic mice result from either pronuclear or cytoplasmic injections of the vector into fertilized eggs. Of 30 mice generated by microinjection, 27 (90%) contained the vector in its episomal form, at less than one copy per cell. This represents a highly efficient means of gene transfer in which the transgene is in a controlled genetic environment. PMID- 3031485 TI - Cloning and characterization of LOS1, a Saccharomyces cerevisiae gene that affects tRNA splicing. AB - Saccharomyces cerevisiae strains carrying los1-1 mutations are defective in tRNA processing; at 37 degrees C, such strains accumulate tRNA precursors which have mature 5' and 3' ends but contain intervening sequences. Strains bearing los1-1 and an intron-containing ochre-suppressing tRNA gene, SUP4(0), also fail to suppress the ochre mutations ade2-1(0) and can1-100(0) at 34 degrees C. To understand the role of the LOS1 product in tRNA splicing, we initiated a molecular study of the LOS1 gene. Two plasmids, YEpLOS1 and YCpLOS1, that complement the los1-1 phenotype were isolated from the YEp24 and YCp50 libraries, respectively. YEpLOS1 and YCpLOS1 had overlapping restriction maps, indicating that the DNA in the overlapping segment could complement los1-1 when present in multiple or single copy. Integration of plasmid DNA at the LOS1 locus confirmed that these clones contained authentic LOS1 sequences. Southern analyses showed that LOS1 is a single copy gene. The locations of the LOS1 gene within YEpLOS1 and YCpLOS1 were determined by deletion and gamma-delta mapping. Two genomic disruptions of the LOS1 gene were constructed, i.e., an insertion of a 1.2 kilobase fragment carrying the yeast URA3 gene, los1::URA3, and a 2.4-kilobase deletion from the LOS1 gene, los1-delta V. Disruption or deletion of most of the LOS1 gene was not lethal; cells carrying the disrupted los1 alleles were viable and had phenotypes similar to those of cells carrying the los1-1 allele. Thus, it appears that the los1 gene product expedites tRNA splicing at elevated temperatures but is not essential for this process. PMID- 3031487 TI - Effect of deletion and insertion on double-strand-break repair in Saccharomyces cerevisiae. AB - I investigated double-strand-break repair in Saccharomyces cerevisiae cells by measuring the frequencies and types of integration events at the PET56-HIS3-DED1 chromosomal region associated with the introduction of linearized plasmid DNAs containing homologous sequences. In general, the integration frequencies observed in strains containing a wild-type region, a 1-kilobase (kb) deletion, or a 5-kb insertion were similar, provided that the cleavage site in the plasmid DNA was present in the host genome. Cleavage at a plasmid DNA site corresponding to a region deleted in the chromosome caused a 10-fold reduction in the integration frequency even when the site was close to regions of homology. However, although the integration frequency was normal even when cleavage occurred only 25 base pairs (bp) outside the deletion breakpoint, 98% of the events were associated not with the usual heterogenote structure, but instead with a homogenote structure containing two copies of the deletion allele separated by vector sequences. Similarly, when cleavage occurred 80 bp outside the 5-kb substitution breakpoint, 40% of the integration events were associated with homogenote structures. From these observations, I suggest that exonuclease and polymerase activities are not rate-limiting steps in double-strand-break repair, exonuclease activity is coupled to the initiation step, the integration frequency is strongly influenced by the amount of homology near the recombinogenic ends, both ends of a linear DNA molecule might interact with the host chromosome before significant exonuclease or polymerase action, and the average repair tract is about 600 bp. PMID- 3031488 TI - [Sonographic diagnosis of solid space-occupying abdominal lesions in childhood]. AB - Between 1981 and May 1986 31 children with solid abdominal tumor masses were observed in our clinic. The first diagnostic procedure was a sonographic examination, followed by further radiological investigations if necessary. 30 cases were examined histologically; in one case the sonographic findings were confirmed by an angiography. The most frequent abdominal masses were neuroblastomas and Wilms tumors (7 cases each). A mesoblastic nephroma was diagnosed in 3 cases, a lymphoma, a hepatoblastoma and a rhabdomyosarcoma 2 times each. One time we found a pancreas carcinoma, a teratoma, a hemangiomatosis of the liver, a malignant Schwannoma, a Ewing sarcoma, an adenoma of the adrenal gland, a pheochromocytoma and an osteosarcoma. According to our own experience and recent reports in the literature it seems possible in most cases, to predict the correct diagnosis of solid abdominal masses using the informations of sonographic imaging. Sonography is a highly specific non-invasive diagnostic tool for planning treatment (e.g. early surgery, cytostatic therapy and/or radiation) of solid abdominal masses. Nevertheless the histological examination should be performed in every case to confirm the definitive diagnosis. PMID- 3031489 TI - [Endemic enterovirus meningitis in 4 newborn infants]. AB - Four infants acquired meningitis within ten days in the same maternity unit. They were not severely ill. Fever was neither high nor long-lasting, and sedimentation rate, leukocyte count and C-reactive protein did not suggest bacterial infection, In contrast, there was a marked pleocytosis in the cerebrospinal fluid in three children with leukocyte counts between 2660 and 5000 cells, predominantly granulocytes. As pathogens enteroviruses were identified. One father and one brother of the infants had mild fever and had identical enteroviruses in their stools, and one other father and two further neonates were unaffected carriers. PMID- 3031490 TI - [Various characteristics of the anti-restriction mechanism in bacteriophage T5]. AB - Bacteriophage T5 is not confined by the restriction systems of the second type EcoRII and EcoRV. Bacteriophage T5 DNA is not modified by EcoRII and EcoRV methylases in vivo. The sites of recognition for restriction endonuclease EcoRV are mapped at 24.4; 57.6; 68.5; 70.2% of T5 DNA, while the sites at 5.1; 7.6% are recognized by EcoRII, the sites at 5.75; 6.0 and 6.5% are recognized by HpaI in FST. A high activity of restriction endonucleases EcoRI and EcoRV is demonstrated in crude extracts of E. coli B834 (RI) and E. coli B834 (RV) cells infected by bacteriophage T5. The simultaneous infection of E. coli B834 (RI) or E. coli B834 (RV) cells by the amber mutants of bacteriophage T5 and the suppressing phage lambda NM761 does not result in the protection of lambda DNA by the T5 anti restriction mechanism. The presented data support the hypothesis that the anti restriction mechanism of bacteriophage T5 is based on prevention of T5 DNA contacts with restriction enzymes by a specific phage protein. PMID- 3031491 TI - [The library of Rickettsia prowazekii genes]. AB - The DNA of Rickettsia provazekii strain E was cleaved by PstI restriction endonuclease under the conditions of partial restriction. The fragments were inserted into the PstI site of pBR325 and cloned in this plasmid. E. coli strain HB101 was used as a recipient for cloning. 880 clones sensitive to ampicillin and resistant to tetracycline were selected from 5120 transformants. The cloning of rickettsial DNA has been confirmed by the blot hybridization technique. Analysis of individual and net probes of the hybrid DNA by gel electrophoresis makes it possible to conclude that 90% of the selected clones harbour hybrid plasmids, the size of the cloned fragments rangers from 0.9 to 10.4 Kb, the obtained library of clones contains 70% of the whole genome of Rickettsia provazekii. PMID- 3031492 TI - [Dynamics of the accumulation of virus particles with different physico-chemical properties in FRhK-4 cells infected with hepatitis A virus]. AB - The propagation time-course of hepatitis A virus (HAV, strain HAS-15) in continuous culture of the foetal rhesus monkey kidney cells (FRhK-4) was investigated. The HAV infectivity and viral RNA content in the infected cells reached the maximal level 5-8 days after infection, while accumulation of hepatitis A antigen (HAAg) continued for 2-3 weeks more. Viral particles with the densities 1.27-1.28 g/cm3 and 1.18-1.22 g/cm3 were isolated from the infected cells as well as the mature virions with the buoyant density 1.33-1.34 g/cm3 in CsCl. The concurrent accumulation of mature virus and "light" particles (1.18 1.22 g/cm3) was registered during infection. Viral particles with the density 1.27-1.28 g/cm3 accumulated predominantly from the 14th to the 21st-24th days after infection. The mature virions (1.34 g/cm3) as well as the particles with the density 1.24-1.25 g/cm3 were isolated from supernatant precipitated by ammonium sulphate. The HAAg activity of both fractions increased progressively in equal proportion in course of infection. PMID- 3031493 TI - [Molecular cloning and study of the expression of a region of a diphtheria toxin gene encoding fragment A of the Streptomyces lividans 66 strain]. AB - The shuttle plasmid pVG202 containing a part of diphtheria toxin gene coding for fragment A has been constructed. S. lividans strain 66 has been transformed by the plasmid pVG202 DNA. The presence of the hybrid plasmid in S. lividans 66 cells determines the production of catalytically active toxoid secreted into the cultural liquid medium. The deleted plasmid pVG205 which determines for the increased catalytic activity has been selected and shown to be stably inherited by the bacterial cells. PMID- 3031494 TI - Mutagenicity of a series of N-alkyl-, N-hydroxyalkyl-, N-haloalkyl- and N carboxyalkyl-N-nitrosoureas in Escherichia coli tester strains: dependence on the uvrA DNA-repair system. AB - A series of N-alkyl-, N-hydroxyalkyl-, N-haloalkyl- and N-carboxyalkyl-N nitrosoureas and some related derivatives were tested for mutagenicity in E. coli B (Arg-) H/r30R (wild-type) and its isogenic Hs30R (uvrA) tester strains. Mutagenic potency in Hs30R, in general, appears to depend on the substituent ( OH, -OCH3, -halogen, -COO- or -COOCH3) on the alkyl group, rather than the chain length or branching of the alkyl group. On the other hand, mutagenic potency in the wild-type H/r30R strain depends on buliness of the substituent and alkyl moiety. The term "uvrA-dependence" of mutation frequency is then defined as the ratio of the mutation frequency in Hs30R versus that in H/r30R at 1 mM dose of mutagens. Its dependence on structure is also discussed. A good correlation was found with the van der Waals volume of the substituted alkyl group, except for compounds having a carboxyalkyl or a branched alkyl group. The carboxyalkyl derivatives are the most weakly mutagenic and most seriously "uvrA-dependent", probably due to the negative charge of the molecule. The possibility of forming epoxides and lactones from N-hydroxyalkyl- and N-carboxyalkyl-N-nitrosoureas, respectively, and their participation in mutagenic potency are discussed. An attempt to correlate the partition property and activation rate of the N nitrosoureas with mutagenic characteristics proved unsuccessful. PMID- 3031495 TI - Photochemical instability of 1-nitropyrene, 3-nitrofluoranthene, 1,8 dinitropyrene and their parent polycyclic aromatic hydrocarbons. AB - The environmental contaminants pyrene, 1-nitropyrene, 1,8-dinitropyrene, fluoranthene, and 3-nitrofluoranthene were exposed to light (greater than or equal to 310 nm) either in DMSO, or following coating onto silica. Under all conditions tested the pyrenyl were less stable than the fluoranthenyl compounds. During irradiation in DMSO or on silica, 1-nitropyrene had half-lives of 1.2 and 6 days, while those of 3-nitrofluoranthene were 12.5 and greater than 20 days, respectively. The photodecomposition of 1,8-dinitropyrene resembled that of 1 nitropyrene with half-lives of 0.7 and 5.7 days. A principle photodecomposition product of 1,8-dinitropyrene was identified as 1-nitropyren-8-ol. It was also found that when the nitroarenes were exposed to light, the loss of compound was associated with a concomitant loss of mutagenicity in Salmonella typhimurium strain TA98. The mechanism of nitrated polycyclic aromatic hydrocarbon decomposition and 1-nitropyren-8-ol formation, and the relevance to the atmospheric disposition of these compounds are discussed. PMID- 3031496 TI - Calcium-activated protease activity in tenotomized muscle. AB - The purpose of this study was to investigate the possible role of calcium activated neutral protease in the disorganization and dissolution of the myofibrils of the rat soleus that occurs following tenotomy. Rats were killed 3, 5, 7, 14, 21, and 42 days after tenotomy of the soleus, and the muscles were removed and assayed for calcium-activated protease activity. Maximal protease activity occurred 1 week after tenotomy, at the time when myofibril organization is completely disrupted. Activity was still high 2 and 3 weeks after the operation, but returned to normal levels by 6 weeks, when muscle histology had returned to normal. The time course of the calcium-activated protease activity corresponded closely to the time course of the morphological changes. Thus, calcium-activated neutral protease may play a major role in myofibrillar proteolysis following tenotomy and in making the myofibril susceptible to proteolytic attack by other, less specific proteases. PMID- 3031497 TI - Calcium-activated protease in hamster cardiomyopathy. AB - A high calcium-requiring protease was purified from the hearts of myopathic hamsters. The biochemical properties of the enzyme were studied with [3H]acetylcasein as substrate. Comparison of the enzyme from hamster and rat hearts indicated no species specificity. Increased levels of the enzyme were associated with the development of cardiac lesions in myopathic hamsters. PMID- 3031498 TI - Studies on the pathogenesis of vincristine-induced neuropathy. AB - The pathogenesis of the peripheral neuropathy induced by vincristine is poorly understood, but interference of vinca alkaloids with microtubule assembly suggests that microtubule changes could be important. This possibility was studied by directly exposing the rat sciatic nerve to graded concentrations of vincristine sulfate. Microtubule length histograms prepared from randomly selected axons showed a unimodal distribution in vincristine and control axons. In vincristine-exposed axons, however, there was a shift to shorter length microtubules, and the mean measured length of microtubules (0.42 +/- 0.37 micron) was significantly (P less than 0.001) shorter than controls (0.67 +/- 0.55 micron). On cross-sections, the vincristine-exposed axons showed a decrease in the number of microtubules per square micrometer of axonal area compared to controls. These findings fit best with a loss of portion(s) of each microtubule and support the possibility that microtubules changes were associated with malorientation of microtubules and neurofilaments, accompanied by free vesicle accumulation and fragmentation of the smooth endoplasmic reticulum. These structural alterations would account for the previously observed abnormalities in axoplasmic transport and would also provide insight into the commonly observed peripheral neuropathy induced by vincristine treatment. PMID- 3031500 TI - Double-step repetitive stimulation in myasthenia gravis. AB - The double-step technique of repetitive nerve stimulation was compared with repetitive nerve stimulation to a distal and a proximal muscle and with single fiber needle electromyography in 10 patients with myasthenia gravis. We conclude that the double-step technique is slightly more sensitive than repetitive nerve stimulation to a proximal muscle but only 60% as sensitive as single fiber electromyography in demonstrating abnormal neuromuscular transmission in myasthenia gravis. PMID- 3031499 TI - A critical review of therapies in acute and chronic inflammatory demyelinating polyneuropathies. AB - Acute and chronic inflammatory demyelinating neuropathies are among the most common treatable neuropathies seen by neurologists. Evidence for effective therapy has only recently been provided by randomized or controlled trials. In the Guillain-Barre syndrome such evidence does not support the use of corticosteroids or immunosuppressive agents. However, when used early in the course, plasma exchange (PE) has been shown to lessen the severity and shorten the duration of the disease; it is indicated only in severely paralyzed patients or those whose rapid deterioration suggests the imminent need for ventilatory support. Some patients with chronic inflammatory demyelinating polyneuropathy (CIDP) respond to corticosteroid therapy or other immunosuppressive agents. PE is also effective in certain patients, but there is no sound evidence to date concerning combined immunosuppression and PE. The rationale of PE in these conditions and whether it is the removal of a toxic factor or the replacement fluid used that is beneficial remains to be clarified. PMID- 3031501 TI - Sensory potentials and sural nerve biopsy: a model evaluation. AB - Compound action potentials (CAPs) were recorded from the sural nerve prior to nerve biopsy in patients with various kinds of polyneuropathy. A previously developed volume conductor model for the CAP is applied to analyze the recorded CAPs in close relation with the morphological findings in the biopsy. First, the fiber diameter histograms obtained from the biopsy are used to simulate CAPs, by assuming a linear relation between fiber diameter and propagation velocity. It is concluded that the simulated CAPs deviate systematically from the recorded CAPs. Next, the assumption of a linear diameter-velocity relation is left, and the assumed fiber velocity distribution is adapted to obtain optimal model reconstructions of the recorded CAPs. It is concluded that the model is capable of reconstructing the recorded CAPs, including slow components and small polyphasic potentials in the case of severe fiber loss. It is demonstrated how the diameter histogram and the optimal velocity distribution can be combined to an empirical estimate of the diameter-velocity relation. PMID- 3031502 TI - Treatment of pityriasis versicolor with itraconazole. PMID- 3031503 TI - Developmental neurobiology. Trophic factor theory matures. PMID- 3031504 TI - Glycine-binding sites and NMDA receptors in brain. PMID- 3031505 TI - Timing and site of nerve growth factor synthesis in developing skin in relation to innervation and expression of the receptor. AB - We show that nerve growth factor (NGF) synthesis in developing skin begins with sensory innervation and that sensory neurons do not express NGF receptors until their fibres reach their cutaneous targets. Both cutaneous epithelium and mesenchyme synthesize NGF, the concentration of messenger RNA for NGF being higher in the more densely innervated epithelium. PMID- 3031506 TI - Rearrangement of a chicken immunoglobulin gene occurs in the lymphoid lineage of transgenic mice. AB - Immunoglobulin (Ig) and T-cell antigen receptor genes rearrange through identical heptamer-nonamer recognition sequences during entry of cells into the B or T lymphoid lineage. A similar enzymatic machinery may be used to perform these highly cell-specific events in these two types of lymphoid cells. We have investigated what the signal may be that triggers the rearrangement of one or other of the receptor genes in B or T cells. Mice from three transgenic lines carrying two, four or twenty copies of the unrearranged chicken lambda light chain locus were analysed. In all three lines the chicken Ig transgene rearranges in B cells; in the line with 20 copies, a rearranged fragment can also be detected in thymus DNA. We conclude that the inserted chicken light-chain locus in its natural configuration contains target sequences that permit specific rearrangement in mouse lymphoid B cells, but that this precise differentiation step may be deregulated in thymic cells when the physiological level of relevant information is experimentally altered. PMID- 3031507 TI - Transfection of the CD8 gene enhances T-cell recognition. AB - Antibodies against CD8 or CD4 antigens can prevent T-cell functions induced by T cell targets. As CD8 or CD4 antibodies can also initiate negative signals in T cells in the absence of appropriate targets it is not clear whether CD8 and CD4 molecules are directly involved in the interaction of T cells with their targets. In previous experiments we have introduced the T-cell receptor alpha- and beta chain genes from a CD8-positive cytolytic T cell specific for the antigen fluorescein (FL) and the H-2D molecule of the major histocompatibility complex (MHC) into a CD8-negative recipient cell. The CD8-positive donor cell lysed both FL-conjugated fibroblasts and lymphoblasts, which express relatively high and low amounts of H-2D molecules, respectively. In contrast the CD8-negative transfectant lysed FL-conjugated fibroblasts only. Here we show that recognition of FL-conjugated lymphoblasts by the transfectant is enhanced by supertransfecting it with the CD8 gene. PMID- 3031508 TI - Fibronectin receptor structures in the VLA family of heterodimers. AB - Multiple cell surface proteins of relative molecular mass 115,000-155,000 (Mr 115K-155K) have been implicated as receptors mediating adhesion to extracellular matrix proteins. But the organization and relatedness of these peptides has remained unclear. In separate studies, the 'very late antigens' VLA-1 (Mr 210K/130K) and VLA-2 (Mr 160K/130K) were initially characterized as surface heterodimers appearing 2-4 weeks after in vitro stimulation of human T cells. Three more VLA heterodimers have since been discovered, which, like VLA-1 and VLA 2, are each composed of unique alpha-subunits in association with a common 130K beta subunit. This paper shows that the common VLA beta-subunit is equivalent to subunits found in structures with known fibronectin and laminin receptor activity, and that VLA-3 and VLA-5 are similar or identical to these previously defined receptors for adhesion molecules. Antibody blocking studies confirmed that at least some of the widely distributed VLA proteins of previously unknown function are involved in cell adhesion to fibronectin and laminin. We suggest that the VLA family of receptors may provide cells with multiple independent substrate adhesion capabilities. PMID- 3031509 TI - Myristic acid is coupled to a structural protein of polyoma virus and SV40. AB - In the lytic cycle of papova viruses, both uncoating of the viral genome after infection and assembly of functional virions take place in the cell nucleus. The mechanisms by which newly internalized virions are targeted to the nucleus and viral DNA encapsidated into particles are poorly understood. Although the major capsid protein VP1 is involved in endocytosis, and largely defines virion structure, the functions of the minor proteins VP2 and VP3 have remained obscure. Here we show that VP2 from both polyoma virus and simian virus 40 (SV40) is covalently linked to myristic acid; this is the first report of a myristylated protein in the nucleus and of a fatty acid being important in the structure of a nonenveloped virus. We consider the implications of this unusual modification on encapsidation and suggest that VP2 may be a scaffolding protein for virion assembly. PMID- 3031510 TI - Genome organization and transactivation of the human immunodeficiency virus type 2. AB - Analysis of the nucleotide sequence of the human retrovirus associated with AIDS in West Africa, HIV-2, shows that it is evolutionarily distant from the previously characterized HIV-1. We suggest that these viruses existed long before the current AIDS epidemics. Their biological properties are conserved in spite of limited sequence homology; this may help the determination of the structure function relationships of the different viral elements. PMID- 3031511 TI - NMDA receptors of dentate gyrus granule cells participate in synaptic transmission following kindling. AB - In the mammalian central nervous system, receptors for the excitatory amino-acid neurotransmitters are divided into three subtypes depending on their sensitivity to three specific agonists: kainate, quisqualate and N-methyl-D-aspartate (NMDA). The ionophores operated by NMDA are gated by Mg2+ in a voltage-dependent manner and allow passage of several cations, including Ca2+ which may be important in plastic alterations of neuronal excitability. Indeed, specific antagonists of NMDA receptors effectively block spatial learning, long-term potentiation and some animal models of chronic epilepsy. Despite their abundance on central neurons, NMDA receptors, with a few noteworthy exceptions, do not generally seem to be involved in low-frequency synaptic transmission. Here we report for the first time that NMDA receptors of the dentate gyrus, where they do not normally contribute to the generation of synaptic potentials, become actively involved in synaptic transmission following long-lasting neuronal changes induced by daily electrical stimulation (kindling) of the amygdala or hippocampal commissures. In contrast to controls, the excitatory postsynaptic potentials (e.p.s.ps) of granule cells in hippocampal slices obtained from kindled animals displayed characteristics typical of an NMDA-receptor-mediated component. The involvement of NMDA receptors in synaptic transmission may underlie the long-lasting changes in neuronal function induced by kindling. PMID- 3031512 TI - An inducible transcription factor activates expression of human immunodeficiency virus in T cells. AB - Human immunodeficiency virus (HIV) production from latently infected T lymphocytes can be induced with compounds that activate the cells to secrete lymphokines. The elements in the HIV genome which control activation are not known but expression might be regulated through a variety of DNA elements. The cis-acting control elements of the viral genome are enhancer and promoter regions. The virus also encodes trans-acting factors specified by the tat-III and art genes. We have examined whether products specific to activated T cells might stimulate viral transcription by binding to regions on viral DNA. Activation of T cells, which increases HIV expression up to 50-fold, correlated with induction of a DNA binding protein indistinguishable from a recognized transcription factor, called NF-kappa B, with binding sites in the viral enhancer. Mutation of these binding sites abolished inducibility. That NF-kappa B acts in synergy with the viral tat-III gene product to enhance HIV expression in T cells may have implications for the pathogenesis of AIDS (acquired immune deficiency syndrome). PMID- 3031513 TI - Direct activation of resting T lymphocytes by human T-lymphotropic virus type I. AB - The mitogenic or antigenic stimulation of T lymphocytes in the presence of accessory cells results in the synthesis of interleukin-2 (IL-2), which, on interacting with de novo synthesized receptors on the membrane, induces the proliferation and differentiation of T cells. T lymphocytes are the preferential target cells of human T-lymphotropic virus type I (HTLV-I). This virus was isolated from leukaemic cells of patients with adult T-cell leukaemia. No viral transcription was detected in these leukaemic cells, indicating that consistent expression of HTLV-I is not needed for maintenance of the neoplastic state. After a short time in culture, these leukaemic cells produce viral particles which immortalize normal T cells. Hence, HTLV-I might be an important agent not only in the development, but also in the initiation, of this lymphoproliferative disease. This possibility was sustained by our previous observations indicating that normal T lymphocytes incubated with HTLV-I are able to form colonies in soft agar, seven days later, in the absence of exogenous IL-2. These results led us to explore further the immediate effects of viral infection on purified resting T lymphocytes. Here, we present evidence that HTLV-I is able to activate T lymphocytes. Indeed, binding of viral particles to these cells induces IL-2 production and IL-2 receptor expression, and triggers T-cell proliferation. This initial activation, which appears to be independent of accessory cells, may be relevant in understanding the role of HTLV-I in the aetiology of adult T-cell leukaemia. PMID- 3031514 TI - The serotonin/noradrenaline-link in brain. I. The role of noradrenaline and serotonin in the regulation of density and function of beta adrenoceptors and its alteration by desipramine. AB - The present studies were undertaken to assess the role of noradrenaline (NA) and serotonin (5HT) in the regulation of the NA receptor coupled adenylate cyclase system and its alteration by desipramine (DMI) in brain structures with or without noradrenergic neuronal projections. In contrast to cortex and limbic forebrain, where chronic DMI administration caused subsensitivity of the NA sensitive adenylate cyclase linked to a down-regulation of beta adrenoceptors, the drug failed to alter the NA receptor coupled adenylate cyclase system in the striatum. Selective lesions of serotonergic axons with 5,7-dihydroxytryptamine caused a significant increase in the density of beta adrenoceptors in cortex, limbic forebrain and striatum and prevented the down-regulation by DMI of beta adrenoceptors in cortex and limbic forebrain while the responsiveness of the NA sensitive adenylate cyclase was reduced to the same extent as in sham-lesioned control animals. The discrepancy between beta adrenoceptor number and NA responsiveness following lesions of 5HT axons was particularly profound in the striatum. The analysis of high- and low-affinity components of agonist binding demonstrated that the increase in striatal beta adrenoceptors is due to a marked increase in receptors with low affinity while the number of receptors with high affinity is unchanged. The results lend further support to the view that the synaptic availability of NA is a prerequisite for the induction of subsensitivity of the NA sensitive adenylate cyclase and for the down-regulation of its beta adrenoceptor population by DMI and that 5HT plays a pivotal role in both the regulation of the number and the function of central beta adrenoceptors. PMID- 3031515 TI - Phorbol-12,13-dibutyrate enhances the cyclic AMP accumulation in rat hippocampal slices induced by adenosine analogues. AB - Phorbol-12,13-dibutyrate (PDiBu) augmented the accumulation of [3H]-cyclic AMP in rat hippocampal slices elicited by the adenosine receptor agonist 5'-(N-ethyl) carboxamidoadenosine (NECA). A similar, but less pronounced, effect was observed when using tetradecanoylphorbol acetate (TPA) provided that the slices were preincubated with the phorbol ester before addition of NECA. PDiBu also induced cyclic AMP accumulation in the absence of NECA, but this effect could be markedly reduced or even abolished by the adenosine antagonist 8-para sulphophenyl theophylline (8-pst). Whereas no clearcut synergism was observed between isoprenaline and PDiBu under normal circumstances, such synergism was observed after elimination of the cyclic AMP accumulation caused by endogenous adenosine. PDiBu slightly enhanced the stimulatory effect of low (0.1-1 microM) concentrations of forskolin, but did not enhance the effect of 10 microM forskolin. In the presence of forskolin, another adenosine analogue, R-PIA, had a biphasic effect on cAMP accumulation. With PDiBu present the inhibitory phase was entirely eliminated and the stimulatory phase substantially enhanced. A synergistic interaction between PDiBu and NECA was found even after inhibition of the Ni-protein by N-ethyl maleimide. It is concluded that the phorbol ester may enhance the stimulatory effect of adenylate cyclase stimulatory compounds, including endogenous adenosine. The effect may partly depend on protein kinase C mediated inactivation of N-protein mediated adenylate cyclase inhibition, but other mechanisms also seem to be involved. PMID- 3031516 TI - Effects of propylthiouracil and methylthiouracil on cyclic AMP and ion movements in rat pancreatic islets. AB - Propylthiouracil and methylthiouracil have been shown to potentiate glucose induced insulin secretion from rat pancreatic islets: the effect of methylthiouracil being less pronounced than that of propylthiouracil. In this study the effects of these substances on cAMP levels, 86Rb+ efflux, 45Ca2+ net uptake, and 45Ca2+ efflux were tested in isolated rat islets in order to obtain information on their possible mechanism of action. Propylthiouracil and to a lesser extent methylthiouracil increased islet cyclic AMP in a concentration related manner. Maximum increases at the highest concentrations tested were 261% and 190% respectively. In the presence of 3 mM glucose propylthiouracil and methylthiouracil led to a decrease in the 86Rb efflux rate. With 5.6 mM glucose, both thiourea derivatives produced an increase in the 86Rb+ efflux rate which was independent of the presence or absence of calcium in the medium. Propylthiouracil and methylthiouracil augmented the 45Ca2+ efflux rate in the presence as well as in the absence of external calcium at various glucose concentrations. Propylthiouracil did not change, and methylthiouracil only slightly augmented, 45Ca2+ net uptake into the isolated islets. It is suggested that the synergistic effect of propylthiouracil and methylthiouracil on glucose-induced insulin release is at least in part due to an increase in islet cAMP levels. Whether the two substances have additional direct effects on ionic fluxes which contribute to their insulinotropic action or whether the observed changes in ion movements are secondary to the elevation of cAMP levels remains to be unclear and needs further investigation. PMID- 3031517 TI - [Atrial natriuretic factor: a versatile hormone produced by the heart]. PMID- 3031518 TI - The immunological activities of components isolated from Corynebacterium parvum in mice injected with polyoma tumor cells. AB - The inhibition of polyoma tumor in CBA mice after immunization with different fractions of Corynebacterium parvum was investigated. Treatment with 200 micrograms of polysaccharide from culture filtrate before s.c. inoculation 2 X 10(6) of tumor cells induced antitumor effect in mice. Treatment with lower doses (1 microgram or 20 micrograms) after transplantation of tumor cells was not effective. Injection of lipopolysaccharide from Escherichia coli abolished a positive response on inhibition of polyoma tumor growth after immunization with Corynebacterium parvum fractions. Additionally, delayed-type hypersensitivity reaction to protein fractions contain tumor antigenic components of membrane vesicles from polyoma tumor cells in the mice immunized with different compounds of Corynebacterium parvum was very strong. These findings suggest that the Corynebacterium parvum preparations are capable to inhibit tumor growth and could be used in special circumstances in immunoprophylaxis. PMID- 3031519 TI - Cyclic AMP levels of C6 glioma cells treated with cis-dichlorodiammine platinum (cis-DDP). AB - Rat C6 glioma cells were cultured for 3-4 days in MEM supplemented with bovine serum. After 10 min incubation of cells with 0.075, 1.0 or 7.5 micrograms ml-1 cis-DDP the basal cAMP levels (7.87 +/- 0.4 pmoles mg-1 protein) were not affected. In the presence of a phosphodiesterase inhibitor, IBMX, an increase of cAMP occurred; the later was more pronounced in cis-DDP treated cells than in the controls. This suggests that both adenylate cyclase and cAMP-phosphodiesterase were proportionally influenced at this period and that the stimulatory effect of cis-DDP on AC could be demonstrated only when increased activity of PDE had been blocked by IBMX. At later time intervals (10 h-40 h), a 5- to 17-fold elevation of cAMP levels was observed even in the absence of IBMX. Pretreatment of the cells with cis-DDP significantly potentiated cAMP accumulation in response to NE alone and to cis-DDP plus NE could be prevented to a large extent by propranolol; in cis-DDP treated cells the propranolol protection was more effective, both in the absence and the presence of IBMX. The pretreatment of cells with an alpha blocker, Regitin, did not significantly influence cAMP accumulation. The results indicate that the cis-DDP stimulated cAMP response to NE is mediated via an interaction with beta-adrenergic receptors. The late increase in cAMP content may be a mediator of the morphological changes in these cells following exposure to cis-DDP. PMID- 3031520 TI - Competition between low- and high-molecular-weight proteins for renal tubular uptake. AB - To explain the occurrence of tubular and glomerular proteinuria in patients with primarily tubular or glomerular dysfunction, it is usually assumed that the mechanisms responsible for the renal tubular transport of small and large proteins are different. The present in vivo study does not support this hypothesis since it clearly shows that small and large proteins (e.g., beta 2 microglobulin and albumin) can compete for renal uptake. Our results lead us to postulate the existence of common tubular reabsorption sites for which proteins exhibit different affinities depending on their charge, size and conformation. PMID- 3031521 TI - Total body potassium in Bartter's syndrome before and during treatment with enalapril. AB - Total Body Potassium (TBK) was measured by whole body counting of 40K in 3 patients with Bartter's syndrome before, after 3 months and after 1 year of treatment with enalapril. In 2 patients TBK was found to be decreased before treatment, whereas TBK was within the normal range in the 3rd. During treatment serum potassium concentration and TBK increased in each subject and symptoms of fatigue and tetany disappeared. Enalapril is shown to be an effective treatment in Bartter's syndrome as it improves serum potassium, TBK and complaints. PMID- 3031522 TI - Spinal evoked responses in the cat. A comparison of different recording techniques with regard to neuromonitoring. AB - Spinal evoked potentials to sciatic nerve stimulation were recorded with epidural electrodes over the dorsal columns. A comparison with potentials to dorsal column stimulation disclosed differences in the typical segmental variation in regard to wave form, amplitude, spinal conduction velocity and conduction time that could be important in neuromonitoring. PMID- 3031523 TI - The effect of increased longevity, produced by dietary restriction, on the neuronal population of area 17 in rat cerebral cortex. AB - Area 17 of the brains of Sprague-Dawley derived rats, maintained on a limited ration of food to maintain their weights at the levels attained by two months of age, was compared with area 17 in control groups of rats fed ad lib. The oldest rats in the diet restricted group were sacrificed at 46 and 48 months of age, by which time their life spans had been extended about 12 months beyond the oldest age that rats fed ad lib achieve, for only few of the latter live as long as 33 months. In this study, the rats which were compared consisted of two groups of ad lib fed rats, one 3 and 6 months of age, and the other 33 months old, and two groups of diet restricted rats, one 26 months old and the other 46 to 48 month old rats (designated as 47 month old rats). Two indices were used to assess whether age affects the volume of area 17. One, the number of clusters of apical dendrites of layer V pyramidal cells per unit area of tangential sections, was the same in all groups, indicating that the lateral spread of area 17 did not alter with age. However, the other index, the thickness of area 17, did change with age, for area 17 was significantly thinner in the 47 month old diet restricted rats than in the other three groups. It was also found that the number of neuronal profiles in strips of sections passing through the entire depth of area 17 is decreased in the 47 month old rats, indicating that neurons had been lost from their cortices. This decrease in the number of neuronal profiles in the 47 month old rats was not due to nuclear shrinkage since the sizes of neuronal nuclei were not significantly different in the older ad lib and diet restricted rats. Determinations of neuronal packing densities in layers II/III, IV, V and VIa suggest that neurons are most frequently lost from the deeper cortical layers of the 47 month old rats, and in these layers large vacuolated spaces, the sizes of neuronal cell bodies, have been encountered. It is suggested that these spaces represent places from which neurons have been lost. It is concluded, therefore, that neurons are lost from area 17 in rats whose longevity is increased by diet restriction. PMID- 3031524 TI - [New knowledge on the calcium pyrophosphate dihydrate (CPPD) crystal deposition disease in the cervical ligamentum flavum]. AB - A case of 64-year-old Japanese woman with cervical radiculomyelopathy caused by the deposition of both calcium pyrophosphate dihydrate (CPPD) and hydroxyapatite crystals in the cervical ligamentum flavum is reported. The patient developed paresthesia in her both hands and gait disturbance in the previous six months. Neurological examination revealed generalized hyperreflexia and hypesthesia in the C7 region. Neuroradiologically, two nodular opacifications were shown in the C3-4 and C5-6 level. CT scan revealed linear and symmetrical calcifications in the C3-4 and nodular calcifications in the C5-6 level. En bloc laminectomy was performed and the patient was discharged from the hospital with improvements of neurological symptoms. Specimens of the spinal laminae and ligamenta flava were studied by light microscopy, scanning electron microscopy (SEM), energy dispersive X-ray microanalysis (X-MA) on SEM, and an X-ray diffraction study was carried out on the crystals. The results showed A. CPPD crystal depositions throughout the ligamenta flava together with a minimal thickening of the ligaments. The deposition was seen in the midzone of the ligaments forming a plate-like distribution and forming lines in the cut surface, the latter coincided well with the CT finding at the C3-4 level. B. The nodule (7 X 6 X 6 mm in size) at the C5-6 level was composed of two crystals, CPPD and hydroxyapatite, the latter being encased by the former. The thickness of the CPPD layer was quite thin varying from 10 to 100 mu. Myelographically, the spinal cord was most severely compressed by the C5-6 nodule. C. Because of the absence of family histories and predisposing factors, the case was considered one of primary or idiopathic CPPD crystal deposition disease. The origin of the deposition of mixed crystals was discussed and it appeared likely that the central part of CPPD crystal deposition had transformed to hydroxyapatite at the C5-6 level. In the literature a considerable number of cases have been reported under the title, "Calcification of the Ligamentum Flavum", and some of them have been shown to have hydroxyapatite deposition. Considering the great similarities in clinical, radiological and histological findings between the so-called "calcification of the lig. fl." and CPPD crystal deposition disease, the present case was the first to clearly indicate that these two conditions belong to a single category, i.e. CPPD crystal deposition disease.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3031525 TI - Excitotoxic mechanisms in hypoglycaemic hippocampal injury. AB - Light and electron microscopy were used to study the effect of hypoglycaemia on selectively vulnerable neurons of rat hippocampus with and without pharmacologic blockade of the N-methyl-D-aspartate (NMDA)-preferring receptor with 2-amino-7 phosphonoheptanoic acid (AP-7). In control hypoglycaemic hippocampi, dark cell change occurs predominantly in dentate granule cells. The topography and ultrastructural appearance of these changes is distinct from that produced by ischaemia or status epilepticus. In hypoglycaemia, mitochondrial calcium accumulation characteristic of ischaemia or status epilepticus is not seen. NMDA receptor blockade markedly attenuates the hypoglycaemic cell injury. Similar attenuation of ischaemic and epileptic brain damage by NMDA receptor blockade suggest that excessive neuronal excitation is a common mechanism of injury in each of the three conditions. PMID- 3031527 TI - Zinc in the anterior pituitary of rat: a histochemical and analytical work. AB - Recent studies have suggested zinc to be a possible modulator of hormone secretion from the anterior pituitary. The three aims of the present study were: to estimate the total amount of zinc in the gland by instrumental analysis; to visualize the zinc at light and electron microscopical levels, and to estimate the amount of zinc that can be visualized by histochemical techniques. PIXE measurement showed the total amount of zinc to be 74 ng/mg dry weight in males and 100 ng/mg dry weight in females, which is a highly significant difference. Histochemically, the zinc was shown by a modified Timm method and the selenium method to be localized to the secretory granules, and to a smaller extent to the Golgi apparatus of the somatotrophs, corticotrophs and thyrotrophs. Chelation of tissue zinc by intravital dithizone treatment effectively blocked subsequent selenium and Timm staining in the secretory granules, and PIXE assays of the chelated metal (extracted into CCl4) showed it constituted less than 5% of the total zinc in the tissue. It is concluded from the study that zinc is present in the anterior pituitary of rats in rather high amounts and that loosely bound zinc, which is suggested to exert a biological effect by itself, can be located to the parts of the cells responsible for production, storing and release of hormones. PMID- 3031526 TI - Noradrenaline, by activation of alpha-1-adrenoceptors in the region of the supraoptic nucleus, causes secretion of vasopressin in the unanaesthetized rat. AB - In unanaesthetized rats chronically prepared with venous and intracerebral cannulae, noradrenaline injected into the region of the supraoptic nuclei caused a dose-dependent increase in plasma vasopressin, measured by radioimmunoassay. A similar response was obtained with phenylephrine, but not with either clonidine or isoprenaline. The secretion of vasopressin was not secondary to change in arterial pressure, since similar injections of noradrenaline resulted in a small increase in arterial pressure, measured in the anaesthetized rat. These results suggest that noradrenaline stimulates alpha-1-adrenoceptors, presumably located on vasopressin-secreting neurones, thereby causing these cells to secrete vasopressin into the circulation. Tyramine injections also resulted in a prompt elevation in plasma vasopressin, indicating that endogenous noradrenaline is capable of releasing vasopressin. PMID- 3031528 TI - Down-regulation of serotonin2, but not of beta-adrenergic receptors during chronic treatment with amitriptyline is independent of stimulation of serotonin2 and beta-adrenergic receptors. AB - Antidepressant drugs down-regulate beta-adrenergic, alpha 2-adrenergic and serotonergic 5-HT2 receptors with a time course that parallels their clinical efficacy, i.e. chronic administration is required (Crews and Smith, 1978; Svensson and Usdin, 1978; Banerjee, Kung, Riggi and Chanda, 1979; Bergstrom and Keller, 1979; Peroutka and Snyder, 1980). In the present study, it was found that the 5-HT2 receptor antagonist, nefazadone (50 mg/kg per day) did not prevent the downregulation of 5-HT2 receptors in the cerebral cortex produced by amitriptyline (10 mg/kg per day), when administered for 3 weeks. Moreover, treatment with nefazadone (50 mg/kg per day) alone for 3 weeks decreased binding to 5-HT2 receptors in cerebral cortex. In contrast, administration of propranolol, the beta receptor antagonist, (10 mg/kg per day) with amitriptyline (10 mg/kg per day) for 3 weeks prevented the down-regulation of beta receptors, but did not alter the decrease in binding to 5-HT2 receptors. In addition, the depletion of central stores of norepinephrine and serotonin by a 4-day treatment with reserpine (5 mg/kg per day) increased binding to beta receptors in the cerebral cortex and hippocampus, but did not affect binding to 5-HT2 receptors in either region. These results suggest that the 5-HT2 receptor is not down regulated by direct stimulation by serotonin agonists and that the down regulation of 5-HT2 receptors by amitriptyline is independent of down-regulation of beta-adrenergic receptors. PMID- 3031529 TI - Down regulation of beta-receptors by bupropion and its major metabolite in mouse brain. AB - Mice were treated with bupropion Compound II (major metabolite of bupropion) or desmethylimpramine (DMI) twice a day intraperitoneally for either 1 or 3 weeks. The binding of dihydroalprenolol and spiroperidol in the frontal cortex and limbic forebrain areas were analyzed. There was a significant reduction in beta receptors in the frontal cortex induced by DMI at both times examined. Bupropion showed a significant reduction of beta-receptor in the frontal cortex by 3 weeks. Though propiophenone did not have any significant effect on beta-receptors in the frontal cortex, it down-regulated beta-receptors in the limbic forebrain area significantly by 1 and 3 weeks. There was no significant effect of buropion or propiophenone on the binding of spiroperidol either in the cortex (S2 receptor) or in the limbic forebrain (dopaminergic). These results show that bupropion may exert part of its clinical effect through its metabolite propiophenone. PMID- 3031530 TI - Age-related change in alpha-adrenergic responsiveness of the urinary bladder of the rat is regionally specific. AB - The effects of age on the responsiveness of the body of the urinary bladder and base of the bladder to alpha-adrenergic agonists were studied. Regions of the bladder were isolated from Fischer 344 rats, ages 7, 16, and 27 months. Maximum isotonic contractions elicited by potassium chloride (KCl) in both regions of the bladder were unaffected by age. In the bladder body there was an age-related increase in the maximum contraction elicited by phenylephrine, norepinephrine and clonidine. No such alteration in responsiveness was observed in the base of the bladder with age. The ED50 values of all three agonists were unchanged with age in both regions of the bladder. The pA2 values of prazosin and yohimbine were approximately 8.5 and 6.0, respectively, in the body of the bladder, and these values were not altered by age. Thus, it is concluded that an age-related increase occurs in the responsiveness of the body of the bladder to alpha adrenergic activation and that these changes are mediated by alpha 1 adrenoceptors. PMID- 3031531 TI - Opioid receptors in the brain of the rat following chronic treatment with desipramine and electroconvulsive shock. AB - The effect of chronic administration of electroconvulsive shock and the antidepressant drug desipramine on binding to opioid receptors was studied in the cerebral cortex, the basal ganglia and the hippocampus of the forebrain of the rat. In vitro, desipramine inhibited the binding of enkephalinamide to opioid receptors through a reduction in the number of binding sites, whereas the affinity was unchanged or only slightly decreased. In vivo desipramine, injected daily (10 mg/kg) for three weeks, did not change the number of opioid binding sites in the forebrain of the rat. Chronic electroconvulsive shock, given twice a week for three weeks, had no effect on the number of opioid binding sites in the forebrain of the rat. The data do not support the hypothesis of shared effects of lithium, tricyclic antidepressants and electroconvulsive shock on the opioid peptide-opioid receptor systems. PMID- 3031532 TI - Formation of sulphidopeptide-leukotrienes in brain tissue of spontaneously convulsing gerbils. AB - Five minutes after the onset of seizures high amounts of immunoreactive prostaglandin (PG) F2 alpha and smaller amounts of sulphidopeptide (SP) leukotriene (LT)-like immunoreactivity could be detected in gerbil brain tissue. Bilateral carotid artery ligation followed by 15 min of reperfusion even more enhanced brain tissue contents of PGF2 alpha and SP-LT-like material. Analysis of the immunoreactive SP-LT-like material by reversed phase high pressure liquid chromatography (h.p.l.c.) revealed immunoreactivity co-eluting with authentic LTC4 and LTD4. PMID- 3031533 TI - Effects of the anticonvulsant taurine derivative, taltrimide, on membrane transport and binding of GABA and taurine in the mouse cerebrum. AB - The effects of a new anticonvulsive derivative of taurine, taltrimide (2 phthalimido-ethanesulphon-N-isopropylamide), and its metabolites on the release, uptake and binding of gamma-aminobutyric acid (GABA) and taurine, were studied in the cerebrum of the mouse. The potassium-stimulated release of taurine from slices of cerebral cortex was inhibited in vitro by taltrimide and its dealkylated metabolite, whereas the stimulated release of GABA was significantly enhanced by both drugs. The uptake of taurine and GABA were not markedly affected. Both taltrimide and its dealkylated metabolite strongly inhibited the sodium-independent binding of taurine to synaptic membranes of brain, the effects on the binding of GABA being less pronounced. The actions on synaptic binding of taurine and on depolarization-stimulated release of GABA may be of significance for their anticonvulsant properties. PMID- 3031534 TI - Selective changes in the in vivo effects of benzodiazepine receptor ligands after chemical kindling with FG 7142. AB - It has recently been demonstrated that kindling occurs with repeated administration of the benzodiazepine "inverse agonist" FG 7142. The present study was an investigation of the effects of other ligands for the benzodiazepine receptor in mice kindled with FG 7142. It was shown that over a range of doses the lowering effects of FG 7142 on the seizure threshold were greater in kindled animals than in control. In contrast, the hypothermic effect of FG 7142 was unaltered. The effects of the partial inverse agonist CGS 8216 were unaltered. The effects of the full inverse agonist DMCM were unchanged except for an enhancement of its convulsant effect when infused at a concentration of 100 mu gm 1-1. Studies with the full agonist benzodiazepine, flurazepam and the full agonist beta-carboline, ZK 93423, showed small but significant reductions in their hypothermic effects. The sedative and anticonvulsant effects of flurazepam were unaltered, whereas the anticonvulsant effects of ZK 93423 were decreased in animals kindled with FG 7142. There was a pronounced reduction in the anticonvulsant and hypothermic effects of the partial agonist beta-carboline, ZK 91296. These data do not fit any simple explanation of kindling being due to a change in the function of benzodiazepine receptors, although they may offer some support for the idea that kindling with FG 7142 produces a change in the effects of all beta-carboline compounds which act at the benzodiazepine receptor. PMID- 3031535 TI - Noradrenergic involvement in catalepsy induced by delta 9-tetrahydrocannabinol. AB - In order to elucidate the role of the catecholaminergic system in the cataleptogenic effect of delta 9-tetrahydrocannabinol (THC), the effect of pretreatment with 6-hydroxydopamine (6-OHDA) or with desipramine and 6-OHDA and lesions of the locus coeruleus were investigated in rats. The cataleptogenic effect of THC was significantly reduced in rats treated with 6-OHDA and in rats with lesions of the locus coeruleus but not in rats treated with desipramine and 6-OHDA, as compared with control rats. On the contrary, the cataleptogenic effect of haloperidol was significantly reduced in rats treated with desipramine and 6 OHDA but not in rats treated with 6-OHDA or in rats with lesions of the locus coeruleus. These results indicate that noradrenergic neurons have an important role in the manifestation of catalepsy induced by THC, whereas dopaminergic neurons are important in catalepsy induced by haloperidol. PMID- 3031536 TI - Naloxonazine and opioid-induced inhibition of reflex urinary bladder contractions. AB - Spontaneous volume-induced contractions of the urinary bladder were recorded isometrically in urethane-anesthetized rats. Contractions were inhibited by alternate submaximal but equieffective doses of the selective mu and delta-opioid ligands [D-Ala2-Me-Phe4,Gly(ol)5] enkephaline (DAGO) and [2-D-penicillamine, 5-D penicillamine] enkephalin (DPDPE), respectively, administered by the intracerebroventricular (i.c.v.) or spinal intrathecal (i.t.) route. Naloxonazine, postulated to be an irreversible mu 1-opioid receptor antagonist, administered by the same route, antagonized the effects of both DAGO and DPDPE. The antagonism of the effect of DAGO was reversed 3-4 hr later but that of DPDPE was more prolonged. Recovery of the effect of DPDPE was observed some 24 hr later. A similar pattern of activity against DAGO and DPDPE given intraventricularly or intrathecally was observed following intravenous injection of naloxonazine (10 mg/kg). Also naloxonazine (i.c.v., i.t. or i.v.) antagonized the effect of morphine given intraventricularly or intrathecally, but antagonism was not observed when morphine was retested 3-4 hr and 24 hr later. Naloxonazine increased the frequency of contraction of the bladder after each route of administration. This effect lasted 1-3 hr and was not seen 24 hr later. Systemic administration of naloxone (10 mg/kg, i.v) also increased the frequency of bladder contraction and attenuated or abolished the effect of DAGO given intraventricularly or intrathecally and the delta-receptor agonist [2-D penicillamine, 5-L-penicillamine] enkephaline (DPLPE).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3031537 TI - Neuropeptides and leukotriene release: effect of peptide histidine isoleucine and secretin in platelet activating factor-stimulated rat lung. AB - We have previously demonstrated that vasoactive intestinal peptide and calcitonin gene-related peptide inhibit leukotriene release from platelet activating factor stimulated rat lung. We now report evidence that Peptide Histidine Isoleucine and Secretin, two naturally occurring peptides which are structurally homologous to vasoactive intestinal peptide, show a significant ability to inhibit leukotriene release in the same animal model. A preliminary hypothesis about the mechanism of this inhibition by the neuropeptides is discussed. PMID- 3031538 TI - Lipolytic potency of proopiomelanocorticotropin peptides in vitro. AB - Four lipolytic active centers were localized in proopiomelanocorticotropin (POMC): in the N-terminal part of POMC ("tryptophane rich peptide"), in the N terminal part of adrenocorticotropic hormone, in the middle portion of beta lipotropin and in the C-terminal part of beta-lipotropin. The weak lipolytic activity of the "tryptophane rich peptide" is not mediated by its two partial sequences gamma-MSH and delta-MSH. The minimal amino acid sequence for obtaining lipolysis from the N-terminal part of ACTH was ACTH 4-10. Lengthening of this amino acid sequence on the N- or C- terminus resulted in a strong increase of lipolytic potency. The minimal effective sequence from the middle portion of beta lipotropin was located in the amino acid residues 47-53 which are identical to ACTH 4-10. Additional amino acids on the N- and C-terminus (beta-lipotropin p 41 58 and beta-lipotropin h 35-56) lead also to increased lipolytic activity. The forth center of POMC resides in the C-terminal part of beta-lipotropin (residues 78-91) because sequences from the N-terminal part of beta-lipotropin 61-91 were ineffective. The order of potency of POMC peptides especially in respect to the minimal effective concentration was ACTH 1-13 (alpha-MSH) greater than beta lipotropin p greater than ACTH greater than ACTH 1-10 greater than beta lipotropin p 61-91. PMID- 3031539 TI - Direct delivery of medication into a brain tumor through multiple chronically implanted catheters. AB - A recurrent malignant glioma was treated with cisplatin delivered directly into the tumor. The drug was delivered via 68 small catheters distributed evenly throughout the tumor area. Each catheter was connected to an osmotic minipump that delivered 0.5 microliter of drug solution each hour for 10 days. There were no side effects from the catheter implantation, chemotherapy, or catheter removal. Although the chemotherapy halted progression of the tumor, it recurred, and the patient died 6 months after treatment. PMID- 3031540 TI - Fiber tracts that contain more opioid binding sites than gray matter does: a quantitative autoradiographic study in the guinea-pig. AB - Opioid binding sites were localized in cryostat sections of guinea-pig brain by in vitro autoradiography using (-)-[3H]bremazocine. Quantification of binding sites in gray matter was accomplished using standard samples of tritium mixed in brain gray matter. The binding sites in fiber tracts were quantified using standards made from white matter to compensate for the quenching caused by myelin. Binding in gray matter corroborated previous findings that the moderately high densities of opioid sites in the cerebellum are of the kappa type, and that the V and VI laminae of the cerebral cortex, the substantia nigra, and olfactory bulb contain high levels of opioid sites in the guinea-pig. Several fiber tracts such as cerebellar white matter and the corpus callosum contained densities of ( )-[3H]bremazocine binding sites equal to, or higher than, the most densely labeled gray matter areas. The dorsal hippocampal commissure and the splenium of the corpus callosum contained 2200 fmol sites/mg protein, two and one half times more than the most densely labeled gray matter areas, the external plexiform layer of the olfactory bulb. These sites may be receptors in transport, but their density constitutes a massive volume of receptors for which a physiological role still needs to be more clearly defined. PMID- 3031541 TI - Kappa opioid receptor activation depresses excitatory synaptic input to rat locus coeruleus neurons in vitro. AB - Intracellular recordings were made from neurons of the rat locus coeruleus contained within a brain slice maintained in vitro. When applied to the slice in known concentrations the selective kappa opioid receptor agonist trans-(+)-3,4 dichloro-N-methyl-[2-(1-pyrrolidinyl)cyclohexyl] benzeneacetamide methane sulphonate (U50488) (0.01-1 microM) produced a concentration-dependent depression of the excitatory post-synaptic potential evoked by electrical stimulation of afferent inputs to the locus coeruleus. This effect was antagonized by naloxone with an apparent dissociation equilibrium constant (Kd) of 28 nM. U50488 did not completely abolish the EPSP. Over the same concentration range U50488 had no effect on the resting membrane potential, input resistance or action potential waveform of locus coeruleus neurons, nor did U50488 depress the depolarization produced by local application of L-glutamic acid. The mu opioid receptor agonists [D-Ala2, NMe Phe4, Gly-ol5] enkephalin (0.003-1 microM) and [D-Ala2, NMe Phe4, Met(O)5] enkephalinol (0.003-1 microM) caused a membrane hyperpolarization concomitant with a fall in neuronal input resistance. These effects were concentration-dependent and antagonized by naloxone with an apparent Kd of 1.5 nM. Mu agonists also caused a depression of the tetrodotoxin resistant action potential. An in vitro autoradiographic study of [3H]bremazocine binding within the locus coeruleus revealed that, although the majority of binding appears to be to mu sites, a significant proportion was displaceable by unlabelled U50488 and thus represented kappa binding sites. The possibility that kappa opioid receptors may be located pre-synaptically within the locus coeruleus, and that activation of these receptors depresses excitatory synaptic input, is discussed. PMID- 3031542 TI - Neural crest-derived proprioceptive neurons express nerve growth factor receptors but are not supported by nerve growth factor in culture. AB - The neural crest-derived, first-order, sensory neurons of the embryonic chick trigeminal mesencephalic nucleus were grown in dissociated, glia-free culture. Whereas brain-derived neurotrophic factor promoted the survival and growth of the majority of these neurons (over 70% after 48 h incubation), nerve growth factor had no effect on their survival. The percentage survival in cultures supplemented with nerve growth factor at concentrations ranging from 0.2 to 625 ng/ml was only 2%, the same percentage survival as in control cultures. Furthermore, nerve growth factor did not change the dose-response of these neurons to brain-derived neurotrophic factor. Although nerve growth factor did not influence the survival of trigeminal mesencephalic neurons in culture, nerve growth factor specifically bound to the great majority of neurons growing in the presence of brain-derived neurotrophic factor. Autoradiographs of cultures incubated with iodinated nerve growth factor showed that the perikarya and processes of neurons were heavily labelled with silver grains. These findings demonstrate the existence of a population of neural crest-derived sensory neurons which express nerve growth factor receptors but are not supported by nerve growth factor in culture. PMID- 3031543 TI - Cytomegalovirus polyradiculoneuropathy in acquired immune deficiency syndrome. AB - A 34-year-old homosexual man with acquired immune deficiency syndrome developed extraocular muscle deficits, chorioretinitis, and paraplegia without sensory symptoms. EMG showed severe diffuse denervation, but only mildly slowed nerve conduction velocities, in both legs. Meningitis persisted for 6 weeks and was exacerbated prior to the patient's death. Necropsy revealed subpial and subependymal cytomegalovirus (CMV) infection. Histology of ventral roots demonstrated proximal CMV infection and massive fiber loss. In this immunosuppressed patient, CMV caused a severe motor polyradiculopathy by selective destruction of the motor neurons of ventral spinal roots and motor cranial nerves. PMID- 3031544 TI - Diagnosis of familial amyloidotic polyneuropathy: isolation of variant prealbumin. AB - A novel, small-scale method was developed for detecting carriers of a prealbumin variant associated with type 1 familial amyloidotic polyneuropathy (FAP). Prealbumin isolated from plasma by a two-step preliminary chromatographic procedure was further separated into two peaks by reverse-phase high-performance liquid chromatography. The normal and variant prealbumins were identified by secondary ion mass spectrometry. The procedure is relatively simple, reliable, and applicable to the definitive diagnosis of FAP in affected patients and also as a preclinical test for the offspring of patients with FAP. PMID- 3031545 TI - Effect of cyanide intoxication on free fatty acid pattern in the brain of rats developing on a diet with various amounts of lipids. PMID- 3031546 TI - [Spontaneous rupture of hepatocarcinoma, a rare cause of hemoperitoneum. Presentation of 2 cases]. PMID- 3031547 TI - [Surgical treatment of parotid tumors. Review of the literature]. PMID- 3031548 TI - [Chemo-radiotherapeutic treatment of non-seminoma germ cell tumors of the testis in the advanced stage]. AB - The results obtained in 18 patients suffering from non seminomatous germ cell tumours of the testis (stage IIC, IID, III), treated at the Division of Radiotherapy-Ospedale Maggiore Novara with chemotherapy and radiotherapy are presented. The radiotherapy technique is described particularly with reference to the use of large fields. CR and PR were obtained in 83.3% and 16.7% of the patients respectively. After a follow up from 13 to 60 months (median 40 months), the total survival is 88.9% and the NED survival is 77.8%. PMID- 3031549 TI - [Pulmonary microcytoma]. AB - 111 patients were treated for lung cancer in 1980-1985. Small cell carcinomas accounted for 9% of the various histological types. The cases classified by TNM were treated by radical surgery where possible. There were no cases of disease free survival at two years. PMID- 3031551 TI - Calcium-dependent neutral protease activity of myelin from bovine spinal cord: evidence for soluble cleavage products of myelin proteins. AB - Myelin basic protein (MBP) is degraded by a calcium-stimulated protease of myelin. An attempt was made to demonstrate soluble endopeptic cleavage products of this reaction. Myelin from bovine spinal cord was incubated at pH 7.5 with 5 mM CaCl2. Protein patterns were evaluated by quantitative polyacrylamide gel electrophoresis. A selective decrease in MBP of residual myelin was accompanied by trace amounts of insoluble cleavage products only. In contrast, the buffer media contained at least 3 distinct peptides of approximate Mr's between 8 and 11 kDa. They comprised approximately 70% of total soluble protein. There was a striking concentration-dependent effect of millimolar CaCl2 on the release of both undegraded MBP and proteolytic polypeptides along with a novel polypeptide of 15 kDa. The results suggest that calcium ions are strongly affecting the retention of loosely bound myelin protein. PMID- 3031550 TI - Quantitative evaluation of alpha- and beta-adrenoceptor modulation of [3H]choline release in guinea pig superior cervical ganglia. AB - [3H] Overflow evoked by 5 min supramaximal preganglionic stimulation at 1 pps has been studied in isolated guinea pig superior cervical ganglion preparations preincubated with [3H]choline. At 15 microM norepinephrine (NE) reduced both the [3H]choline overflow and endogenous acetylcholine release by 59.4 and 54.1% respectively; the dose-response curve for NE inhibitory action is described. Evidence is given that endogenous catecholamines effectively reduce ACh release from the ganglia. After blocking the inhibitory action of endogenous NE, a significant beta-adrenoceptor-mediated facilitatory effect on ACh release could be observed. Preincubation of the ganglia with different combinations of alpha 1 and alpha 2 agonists (phenylephrine, 10 microM and clonidine, 1 microM respectively) and antagonists (prazosin, 10 microM and yohimbine, 3 microM) showed that the adrenoceptors involved in alpha-mediated NE inhibition of ACh output are exclusively of the alpha 2-type. PMID- 3031552 TI - Inhibition of neuronal Cl-stimulated Mg-ATPase by alpha-human atrial natriuretic polypeptide. AB - Effects of alpha-human atrial natriuretic polypeptide (alpha-hANP) on neuronal membrane-bound adenosine triphosphatases (ATPases) (Na, K-ATPase, Cl-stimulated Mg-ATPase and anion-insensitive Mg-ATPase) were examined using rat brain microsomes. This peptide inhibited Cl-stimulated Mg-ATPase in a dose-dependent manner. The inhibition was non-competitive with respect to ATP, and was not affected by Cl-, the most potent stimulator of this enzyme. The activities of Na,K-ATPase and anion-insensitive Mg-ATPase were not changed by alpha-hANP at all. PMID- 3031553 TI - Modulation of beta-adrenergic receptors in the pituitary gland following adrenalectomy in rats. AB - The effects of adrenalectomy on beta-adrenergic receptors in the rat pituitary were examined using quantitative in vitro autoradiography with 125I iodocyanopindolol (125ICYP). 125ICYP binding in the anterior, intermediate and posterior lobes of the pituitary gland was significantly increased in chronically adrenalectomized rats. The increase in 125ICYP binding sites in the rat pituitary following adrenalectomy was not reversed by glucocorticoid replacement with dexamethasone. These data indicate that catecholamines of adrenomedullary origin are capable of modulating beta-adrenergic receptors in the pituitary gland and suggest that peripheral epinephrine may be important in regulating pituitary hormone secretion. PMID- 3031555 TI - Evidence for the presence of GABAergic and glycine-like systems responsible for cardiovascular control in the nucleus tractus solitarii of the rat. AB - Microinjections of gamma-aminobutyric acid (GABA) and glycine, into the medial area of the nucleus tractus solitarii (NTS) of the rat, led to an increase in arterial pressure and heart rate. The GABA receptor antagonist bicuculline and the glycine receptor antagonist strychnine decreased both of these cardiovascular parameters whereas the GABA uptake inhibitor nipecotic acid produced hypertension. High K+ stimulation caused a calcium-dependent release of GABA and glycine from tissues in the area of the NTS. Our results suggest that GABA and glycine may modulate the cardiovascular control within the NTS. PMID- 3031554 TI - Evidence for basal and stress-induced release of corticotropin releasing factor in the push-pull cannulated median eminence of conscious free-moving rats. AB - Fourteen adult male rats were successfully implanted in their median eminence with a push-pull cannula perfusing an artificial fluid at a rate of 13 microliters/min, to measure the release of corticotropin releasing factor (rCRF 41) under physiological conditions assessed by baseline plasma adrenocorticotropic hormone (ACTH) levels. In the basal conscious free-moving state, rCRF-41 release displayed a fluctuating pattern, with peaks about every 45 min at a mean value of 9.0 +/- 0.7 pg/15 min sample (n = 42) vs a mean trough value of 4.1 +/- 0.3 pg/sample (n = 44). Ether stress was followed by a striking rise in rCRF-41 release which generally lasted about 45 min and reached mean peak values of 54.3 +/- 3.2 pg/15 min sample (n = 7). These data constitute the first direct measurements of basal and stress-induced CRF releases in conscious unrestrained rats. PMID- 3031556 TI - Loss of hippocampal [3H]TCP binding in Alzheimer's disease. AB - We have previously demonstrated a marked loss in N-methyl-D-aspartate (NMDA) receptors in the hippocampus and cerebral cortex of patients dying with dementia of the Alzheimer type (DAT). In addition, we have found that the dissociative anesthetic N-(1-[2-thienyl]cyclohexyl)3,4-piperidine ([3H]TCP) binds to a site whose regional distribution is highly correlated with that of NMDA receptor sites. We studied the binding of [3H]TCP to sections of hippocampi from 8 controls, 12 patients with DAT and 7 patients with other dementias. [3H]TCP binding was significantly reduced in strata pyramidalia of CA1/CA2, CA3 and subiculum of DAT hippocampal formation compared to that of control. Labelled dissociative anesthetics could potentially be used with positron emission tomography in the diagnosis of DAT. PMID- 3031557 TI - 2,5-Hexanedione neuropathy is associated with the covalent crosslinking of neurofilament proteins. AB - Protein composition in different segments along nerves from rats intoxicated with 2,5-hexanedione (HD) was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis and by immunoblotting, using monoclonal antibodies specific for each of the three neurofilament polypeptide components (H, M, and L). Comparison with nerve protein extracts from normal (control) rats revealed a disappearance of the largest neurofilament polypeptide (H), accompanied by accumulation of higher-molecular-weight products that were immunoreactive with H-specific antibodies. Both the extent of this crosslinking and its localization in particular portions of peripheral nerves showed a correlation with HD dosage and with the known progression of ultrastructural features during HD-induced neuropathy. Similar changes were not detected for the M and L neurofilament components. PMID- 3031558 TI - Effect of lead on Na+,K+ATPase activity in the developing brain of intra-uterine growth-retarded rats. AB - Lead (Pb) intoxication in developing mammals, including humans, produces serious brain damage. In addition, it is known that nutritional status influences the susceptibility to Pb toxicity. We developed an in utero undernutrition model based on restriction of blood supply to fetuses on d 17 of pregnancy (IUGR rats). The aim of this study was to investigate in vitro the possible effect of Pb on Na+, K+ATPase activity in the brain of developing IUGR and control rats from 6 to 60 d after birth. In addition, we measured the stimulation of Na+, K+ATPase by the monoamines noradrenaline and serotonin. Our results show that: The neurotoxic effect of Pb is an age-related phenomenon. Both IUGR and control rats were more sensitive to Pb in the first week of life. In adults, Pb had a weak inhibitory potency; the delayed matured brain in IUGR animals seemed less sensitive to Pb when compared to age-paired control rats; in the IUGR group, at 15 and 22 d, low doses of Pb had a stimulatory effect on Na+, K+ATPase instead of an inhibitory effect; noradrenaline and serotonin stimulated Na+, K+ATPase activity to an equivalent extent, but this was greater in IUGR than control rats; and at low Pb concentrations, the studied monoamines reversed Pb-induced inhibition. PMID- 3031559 TI - Development of adrenergic receptor binding sites in brain regions of the neonatal rat: effects of prenatal or postnatal exposure to methylmercury. AB - In order to understand the effects of developmental exposure to methylmercury on the ontogeny of synaptic function, we examined the impact of prenatal or postnatal exposure on acquisition of receptor binding sites for norepinephrine. The actions of the mercurial were both regionally- and receptor subtype-selective and depended upon the maturational profile of each region. alpha 1- alpha 2- and beta-receptor sites in the cerebellum, the region which develops last, were the most vulnerable to methylmercury. In contrast, the same receptor subtypes in the midbrain + brainstem, which develops earliest, were resistant to methylmercury. The cerebral cortex, which matures at a time midway between cerebellum and midbrain + brainstem, also displayed intermediate vulnerability to actions of methylmercury on receptors. Within the cerebellum, prenatal exposure to 1 mg/kg of methylmercury interfered the most with ontogeny of alpha 1-receptor binding, less so for alpha 2-receptors and least for beta-receptors. Lower doses of methylmercury tended to increase receptor binding for all subtypes, a fact which may contribute to promotion of neurological development seen in animals exposed to those levels. These data support the view that methylmercury exerts a dual spectrum of action on developing synapses, with promotional effects predominating at low doses and inhibitory actions at higher doses. The net effect on responsiveness to neurotransmitters is influenced, in part, by the actions on developing receptor sites which are in turn dependent upon the specific regional timetables for cellular maturation and receptor acquisition. PMID- 3031560 TI - Neurotoxicants and central catecholamine systems. AB - The ubiquitous functional roles of brain catecholamines have led to the notion that in vitro neurochemical changes in these systems may predict neurotoxicity. Conversely, others have argued that the appropriate use of neurochemical methods is for testing specific hypotheses that are developed based on observed phenomena. Three studies from this laboratory are presented in support of the latter hypothesis. The first example is with inorganic lead, a major environmental pollutant. The effects of small doses of lead on CNS development have been difficult to quantify or study mechanistically. However, the serendipitous finding that lead exposure during early postnatal development increased lithium-induced polydipsia (LIP) has provided clues that have permitted testing of specific neurochemical hypotheses related to dopamine systems. Conversely, the administration of either trimethyl- or triethyltin to rats during perinatal periods causes profound neurotoxicity. However, although some changes in the neurochemistry of catecholamine systems have been found, these data have provided little insight into either the cause or sequelae of toxicity. Finally, the food color erythrosin (FD & C Red #3) was hypothesized to be a neurotoxicant because it disrupted neurotransmitter uptake in vitro. Our data suggested this was an artifact of the methodology, a position supported by clinical and behavioral studies. These data provide examples of the strengths and weaknesses in neurochemical approaches to neurotoxicity. PMID- 3031561 TI - Function of glycoprotein carbohydrate. PMID- 3031562 TI - Biochemical evaluation of organophosphorus-ester induced delayed neurotoxicity (OPIDN): the role of 2',3'-cyclic nucleotide 3'-phosphohydrolase. PMID- 3031563 TI - Effect of mercury on rabbit myelin CNP-ase in vitro. AB - 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNPase) catalyzes hydrolysis of 2',3'-cyclic nucleotides to form the corresponding 2'-monophosphates. Rabbit myelin fraction with CNPase specific activity between 30-40 mumoles/min/mg protein was incubated in the presence of various inorganic and organic heavy metal compounds: HgCl2; (CH3Hg)OH; Pb(NO3)2; Pb(C2H302)2.3H20; (C2H5)2Pb; (C2H5)3SnCl. The enzyme has been shown to be almost exclusively sensitive to mercurials in microM concentration range. This would arise from the high solubility of mercurials in organic solvents, which allows them to penetrate into hydrophobic regions of the enzyme to react with active sulfhydryl groups. CNP-ase inhibition by methylmercury was biphasic: A reversible, non-competitive inhibition with an apparent Ki = 1 microM occurred after a 5 min preincubation time of the enzyme with the inhibitor. In the case of longer preincubation time, as well as in the presence of HgCl2, the graph of enzyme activity versus protein concentration intercepted the abscissa to the right of the origin, indicating that mercurials are irreversible inhibitors of the enzyme. After 45 min of preincubation of the inhibitors with the enzyme 1 nmol of HgCl2 completely blocks CNP-ase activity equivalent to 15.6 micrograms of myelin protein, whereas 1 nmole of Met-Hg blocks activity in 9.9 micrograms proteins. This apparently irreversible inhibition of CNP-ase activity by HgCl2 could be fully restored by the use of an excess of hydrophobic low molecular weight thiols, lipoic acid being the most efficient. Dithiothreitol, a hydrophilic complexing agent, was potent to reverse the inhibition caused by Met-Hg only during the short time experiments. Both low molecular weight thiols, and also EDTA in the case of inorganic mercury could prevent the inhibition of CNP-ase by mercurials, if preincubated for 15 min with the inhibitors, prior to the addition of the enzyme. The irreversible type of inhibition of CNP-ase by Met-Hg was only partially reversed in the presence of low molecular weight thiols. This suggests that the formation of a metal-mercaptide complex is not the only mechanism of inhibition by methylmercury. The possibility of lipid peroxidation triggered by methylmercury with subsequent inhibition of the enzyme activity was not supported by the experimental results. In fact, myelin associated CNP-ase activity appears to be very resistant to the structural membrane alterations caused by lipid peroxidation. PMID- 3031564 TI - Brain neurotransmitter receptor alterations in offspring of rats exposed to phenobarbital, phenytoin or their combination during pregnancy. AB - Neurotransmitter receptor binding was studied in the progeny of rats exposed to phenobarbital (80 mg/kg), phenytoin (80 mg/kg) or their combination daily from 5 to 20 days of gestation. At 22 days postnatally, female pups in all treated groups showed decreased binding of [3H]spiroperidol to striatal membranes. A decreased binding of [3H]diazepam to frontal cortical membranes was observed in the female pups of rats exposed to phenytoin or the combination. The Scatchard analysis of data from the affected groups revealed a decrease in the maximum number of dopamine and benzodiazepine receptor sites and no change in their affinities. The binding of [3H]spiroperidol or [3H]diazepam was not altered significantly in any of the drug-treated male pups. Also, no significant change in the binding of [3H]serotonin or [3H]quinuclidinyl benzilate to frontal cortical membranes was observed in any of the treated groups. At 60 days postnatally, no significant change in the binding of any of the above ligands was evident. The above prenatal drug exposures appear to cause transitory subsensitivity of dopamine and benzodiazepine receptors in rats. PMID- 3031566 TI - [Radioisotope study of reparative osteogenesis in the treatment of transcondyloid fractures of the humerus (experimental study)]. PMID- 3031565 TI - Acute viral hepatitis in Auckland. AB - A community survey was undertaken in order to determine the seroepidemiologic pattern of acute viral hepatitis in Auckland. As hospital records and Health Department notifications underestimate the problem, all patients with a serum alanine transpeptidase (ALT) level of greater than 200 mu/l (X 5 normal) were investigated for viral liver disease. Over a four month period a total of 303 cases of acute viral hepatitis were identified, 49 (16.2%) were hepatitis A, 11 with coincident hepatitis B, 88 (29%) were hepatitis B, 80 (26.4%) nonA nonB hepatitis, 81 (26.7%) Epstein Barr virus hepatitis and 5 (1.6%) cytomegalovirus hepatitis. Hepatitis A and hepatitis B occurred with increased frequency among Maoris and Polynesians, while Epstein Barr virus, cytomegalovirus and nonA nonB hepatitis occurred more frequently among Europeans. The incidence of acute symptomatic viral hepatitis (excluding cytomegalovirus and Epstein Barr virus infections) was 78 cases/10(5)/per year for the Auckland region in this survey. PMID- 3031567 TI - [Targeting of anthracyclines and hepatomas]. AB - DNR can be targeted to the liver by linking to galactosylated human serum albumin (AG). The linkage is stable in the blood stream and allows the release of active DNR after endocytosis of the conjugate by the target cells. Receptors for AG are present at the cell membrane of hepatocytes, primary human hepatoma cells and lung metastases. After i.v. administration of AG-DNR more than 70% of the drug is taken up by the liver and in rats 50% of DNR is eliminated in the bile after 16 h. Nude mice bearing human hepatoblastoma cells implanted s.c. were treated twice a week with a dose equivalent to 6.6 mg/kg of DNR either for the free drug or the conjugate AG-DNR. After 7 injections, tumor growth is inhibited in both case, however, the conjugate seems more active and is at least twice less toxic in terms of LD50. A phase I clinical trial of AG-DNR on 11 patients bearing hepatocarcinoma revealed that despite a transitory hyperthermia (37-38 degrees C) during the first day of the 4 day-perfusion, and modifications of hepatic enzymes in 3 cirrhotic patients, no hematologic and cardiac toxicity could be detected. A subjective response has been obtained in half of the patients with a decrease of plasmatic alpha-foetoprotein levels by more than 50% in 4 patients and one complete remission of more than 23 months with disparition of pulmonary metastases. PMID- 3031568 TI - [Effect of SIN-1 on the synthesis of leukotrienes in human polymorphonuclear cells]. AB - Leukotriene (LT) synthesis triggered with ionophore A23187 or inflammatory stimuli (PAF, fMLP) in human polymorphonuclear leukocytes (PMNL) was markedly decreased by SIN-1, an active metabolite of molsidomine. This inhibition occurred in the absence or in the presence of exogenous arachidonic acid (AA). These results suggest that SIN-1 inhibited both the release of AA from membrane phospholipids and the 5-lipoxygenase activity in human PMNL. Molsidomine was without any effect on LT formation. PMID- 3031569 TI - [Mechanism of the vasodilating effect and blood platelet- antiaggregating activity of molsidomine and SIN-1]. AB - Present evidence indicates that the active metabolite SIN-1 of the prodrug molsidomine dilates vascular smooth muscle and inhibits platelet aggregation by a direct stimulatory effect on soluble guanylate-cyclase in the cytosol of vascular smooth muscle cells and platelets, respectively. Evidence from studies in bovine coronary arteries is presented to proof the causal relation between SIN-1 induced rises in cGMP and relaxation under a variety of pharmacological conditions. In contrast to organic nitrates, SIN-1, which activates guanylate-cyclase in vitro independently of the presence of added cysteine, does not cause tolerance. Tolerance most pronounced with nitroglycerin appears to be due to a direct inactivating effect of this drug on guanylate-cyclase. PMID- 3031570 TI - [Dynamics of succinate dehydrogenase and cytochrome oxidase activity in the myocardium and brain of hypokinetic rats]. PMID- 3031571 TI - [Mechanism of the depression of the heart pumping function in experimental myocardial infarct]. PMID- 3031573 TI - [Nocturnal intragastric administration of phosphates in the treatment of familial hypophosphatemic rickets]. PMID- 3031572 TI - [A case of intravascular bronchiolo-alveolar tumor (IVBAT)]. PMID- 3031574 TI - Neonatal rotavirus vaccination with RIT 4237 bovine rotavirus vaccine: a preliminary report. AB - We vaccinated 244 newborn infants orally with RIT 4237 bovine rotavirus vaccine or placebo and followed them serologically and clinically for 16 months. Initially 39 of the 119 (33%) vaccine recipients compared with 1 of the 120 placebo recipients seroconverted by enzyme-linked immunosorbent assay immunoglobulin M. After the first winter rotavirus season, at 7 months of age 55% of the vaccinated infants and 37% of the unvaccinated infants were rotavirus seropositive by enzyme-linked immunosorbent assay-immunoglobulin G (P less than 0.01, chi square test). At 12 months of age, after a low rotavirus prevalence season, 34% of the vaccinated children and 23% of the unvaccinated children remained seropositive. There were 14 confirmed episodes of rotavirus gastroenteritis in the vaccine group and 10 episodes in the placebo group during the first 16 months. However, only 1 of the episodes in the vaccine group was severe, 4 were moderately severe and 9 were mild, whereas 7 episodes in the placebo group were severe and 3 were moderately severe (P less than 0.001 between groups, Fisher's exact test). There was no clear correlation between vaccine induced clinical protection and initial serologic response (enzyme-linked immunosorbent assay-immunoglobulin M) to vaccination, but during follow-up severe rotavirus gastroenteritis was more likely to occur in children with no serum rotavirus immunoglobulin G antibody at the time of infection. We conclude at the present stage that neonatal rotavirus vaccination with RIT 4237 vaccine gives no protection against rotavirus infection but appears to modify the severity of gastroenteritis. PMID- 3031576 TI - Parainfluenza type 3 in newborns. PMID- 3031575 TI - Longitudinal study of rotavirus infection and gastroenteritis in families served by a pediatric medical practice: clinical and epidemiologic observations. AB - During 29 months of prospective longitudinal study of diarrhea in the home, human rotaviruses (HRVs) infected one or more members in 51% of 65 families, 35 of 126 children (28%) and 16 of 124 adults (13%). Within the 33 affected families, 57% of 62 children and 25% of 65 adults were infected. HRV gastroenteritis peaked at 40/100 person years at ages 12 to 23 months and decreased to 5 episodes/100 person years in adults. Among 25 children 0 through 36 months of age who had HRV infection, 88% were symptomatic. Of the 22 children with symptomatic HRV infection, 1 required hospitalization and 8 were seen by their physician for supportive care. HRVs were found in 12% of 216 stools obtained during gastrointestinal illness, but in only 0.2% of 1238 non-illness stools tested. HRV infections were noted as early as October and as late as April. Of 33 families who were studied for 2 seasons, at least 1 individual in each of 3 families experienced HRV infections in both years, but only one, an adult, shed virus and had symptoms in both seasons. PMID- 3031577 TI - Intravenous hyperimmunoglobulin to prevent varicella-zoster infection. PMID- 3031578 TI - Effect of water intake on Na-K-ATPase in nephron segments of the desert rodent, Jaculus orientalis. AB - Na-K-ATPase activity was measured in individual pieces of nephron microdissected from collagenase-treated kidneys of jerboas, Jaculus orientalis. Na-K-ATPase activity was high in the distal convoluted tubule, intermediate in the thick ascending limb of the loop of Henle and low in the proximal and collecting tubule. When jerboas were adapted for several weeks to a hydrated diet and excreted a more diluted urine, Na-K-ATPase activity was altered in specific segments of the nephron: 1. In the proximal convoluted tubule, Na-K-ATPase activity decreased, especially in the juxtamedullary nephrons, suggesting that internephron heterogeneity was diminished; 2. In the medullary thick ascending limb, but not in the cortical portion, Na-K-ATPase activity decreased by 30%; 3. Na-K-ATPase was also diminished in the cortical collecting tubules (by 20%) but not in the medullary collecting tubule. Morphometric measurements also indicate that changes in Na-K-ATPase activity observed in the thick ascending limb are correlated to a cell atrophy, whereas in the collecting tubule, they occur independently of any visible morphological alteration. These differences in Na-K ATPase activity are likely to be secondary to the changes in the plasma concentration of vasopressin previously described during such adaptation and to be involved in the control of water and sodium handling. PMID- 3031579 TI - Pertussis toxin inhibits negative inotropic and negative chronotropic muscarinic cholinergic effects on the heart. AB - We injected rats with pertussis toxin, known to cause ADP ribosylation of the Gi regulatory protein of the adenylate cyclase complex and of another closely related GTP binding protein in the heart, and after 7 days we examined several effects of muscarinic activation on the heart. The negative chronotropic effect of carbamoylcholine on spontaneously beating perfused hearts was conspicuously diminished. While 10(-5) mol/l carbamoylcholine invariably produced heart arrest in control rats, the heart rate did not decrease by more than 20% in the toxin treated rats even when the concentration of carbamoylcholine was raised to 10(-2) mol/l. The negative inotropic effect of carbamoylcholine examined on electrically paced ventricles perfused with isoproterenol was reduced, while the maximum positive inotropic effect of isoproterenol was substantially increased after toxin treatment. The inhibitory action of carbamoylcholine on the isoproterenol stimulated accumulation of cyclic AMP in the heart auricles was attenuated. The weakening by pertussis toxin of the negative inotropic effect of carbamoylcholine is probably mainly due to the ADP ribosylation of the Gi regulatory protein and the subsequent loss of influence of muscarinic receptors on adenylate cyclase. The blockade of the negative chronotropic action of carbamoylcholine by pertussis toxin strongly indicates, together with other recently published evidence, that the Gi or another closely related GTP binding protein in the cardiac pacemaker cells is involved in the coupling of muscarinic receptors to the K+ channels. PMID- 3031580 TI - Event-related cross-correlations between spike trains illustrated on interactions between motor units and muscle spindle afferents. AB - A method is presented for computing correlation coefficients of two (or more) output spike trains in temporal relation to one (or more) input even trains. These event-related correlation functions are computed by convolving the output spike trains, represented as point processes, with rectangular pulses of selectable width, and by then calculating linear correlation coefficients for the pairs of amplitude values obtained from the two convolved processes in temporal relation to the input events. The merits of this technique are illustrated on stimulus trains delivered to motor units (MUs) and output spike trains recorded from muscle spindle afferents of the same cat hindlimb muscle. The correlation functions obtained show the temporal course of the correlated firings of the two afferents (mostly Ia afferents from primary muscle spindle endings) as a function of time from MU activation; they are compared with the conventional cross correlation histograms (CCHs) between afferents and with peri-stimulus time histograms (PSTHs) between stimulus and afferent firing patterns. Stimulus related cross-correlation functions as displayed here can be calculated for any three spike trains. Possible extensions of the method to larger numbers of input and output channels are also discussed. PMID- 3031581 TI - Cloning and characterization of the gene coding for cytoplasmic seryl-tRNA synthetase from Saccharomyces cerevisiae. AB - We have screened a Saccharomyces cerevisiae expression library with antibodies against seryl-tRNA synthetase (SerRS) from baker's yeast. In this way we obtained clones which contain serS, the structural gene for seryl-tRNA synthetase. Genomic Southern blots show that the serS gene resides on a 5.0 kb SalI fragment. Nucleotide sequence analysis of the genes revealed a single open reading frame from which we deduced the amino acid sequence of the enzyme consistent with that of two peptides isolated from SerRS. The enzyme is comprised of 462 amino acids consistent with earlier determinations of its molecular weight. The codon usage of serS is typical of abundant yeast proteins. Nuclease S1 analysis of serS mRNA defined the RNA initiation site 20-40 bases downstream from an AT rich sequence containing the TATA box and 21-39 nucleotides upstream of the translation initiation codon. Yeast strains transformed with the cloned gene overproduce seryl-tRNA synthetase in vivo. PMID- 3031582 TI - Human dihydropteridine reductase: characterisation of a cDNA clone and its use in analysis of patients with dihydropteridine reductase deficiency. AB - Deficiency of human dihydropteridine reductase (hDHPR) causes malignant hyperphenylalaninemia. We report the isolation of a cDNA clone for hDHPR that spans the complete coding region, and present the nucleotide sequence and the predicted amino acid sequence. The hDHPR protein does not share extensive homology with the enzymatically related protein human dihydrofolate reductase. Patients with hDHPR deficiency were analysed for the presence of hDHPR cross reacting protein, mRNA encoding hDHPR, and chromosomal DNA rearrangements. The results show that this inherited error of metabolism can result from a variety of mutations. However, no major rearrangements were seen in 11 patients analysed by Southern blotting. Three RFLPs were found with the restriction endonucleases AvaII and MspI. These RFLPs are useful for prenatal diagnosis of hDHPR deficiency. PMID- 3031583 TI - The two classes of genes for the major potato tuber protein, patatin, are differentially expressed in tubers and roots. AB - The major potato tuber protein, patatin, is a family of 40kd glycoproteins that constitutes forty per cent of the soluble protein in tubers but is generally undetectable in other tissues. Fused rocket immunoelectro-phoresis was used to detect in roots patatin that is immunologically different from tuber patatin. Western blots of SDS-polyacrylamide gels show root patatin to have a different molecular weight distribution than tuber patatin isoforms, but immunoprecipitation of in vitro translation products shows the patatin precursors to be of similar molecular weight in both tissues. This suggests that post translational processing may differ in tubers and roots. Northern blots show that tuber and root patatin mRNAs are of similar size, but tuber transcripts are about 100-fold more abundant. 5' S1 nuclease and primer extension mapping suggests the class of patatin transcripts expressed in roots (class II transcripts) to be a subset of patatin transcripts expressed in tubers (classes I and II). Class II patatin mRNAs differ from class I transcripts by the presence of a 22 nucleotide insertion just upstream of the initiation codon. These data demonstrate that expression of the patatin multigene family is differentially regulated in tubers and roots. PMID- 3031584 TI - A combination of RNase H and S1 nuclease circumvents an artefact inherent to conventional S1 analysis of RNA splicing. AB - S1 nuclease mapping is commonly used to analyze transcription and processing of unlabelled RNAs. However, the S1 protocol that appears best suited to demonstrate splicing of a particular RNA (using an intronless probe that is 5' end-labelled in the downstream exon) is not diagnostic as expected. Rather, both intron containing RNA and intronless RNA confer protection of probe across the splice juncture. To unambiguously demonstrate correctly spliced RNAs that begin at a specific initiation site, we present a procedure in which unspliced RNA molecules are first cleaved by RNase H following annealing to an intronic DNA fragment and the remaining RNA is then subjected to S1 analysis using an intronless probe present in vast excess. Only spliced, correctly initiated transcripts can protect the probe across the splice junction and up to residue +1. This RNase H/S1 method provides a broadly applicable technique with which to demonstrate splicing and initiation of a variety of transcripts, especially ones from transfected genes that can arise both from the normal and from activated cryptic initiation sites. PMID- 3031585 TI - Characterization of cDNA clones for human myeloperoxidase: predicted amino acid sequence and evidence for multiple mRNA species. AB - Myeloperoxidase is a component of the microbicidal network of polymorphonuclear leukocytes. The enzyme is a tetramer consisting of two heavy and two light subunits. A large proportion of humans demonstrate genetic deficiencies in the production of myeloperoxidase. As a first step in analyzing these deficiencies in more detail, we have isolated cDNA clones for myeloperoxidase from an expression library of the HL-60 human promyelocytic leukemia cell line. Two overlapping plasmids (pMP02 and pMP062) were identified as myeloperoxidase cDNA clones based on the detection with myeloperoxidase antiserum of 70 kDa protein expressed in pMP02-containing bacteria and a 75 kDa polypeptide produced by hybridization selection and translation using pMP062 and HL-60 RNA. Formal identification of the clones was made by matching the predicted amino acid sequences with the amino terminal sequences of the heavy and light subunits. Both subunits are encoded by one mRNA in the following order: pre-pro-sequences--light subunit--heavy subunit. The molecular weight of the predicted primary translation product is 83.7 kDa. Northern blots reveal two size classes of hybridizing RNAs (approximately 3.0-3.3 and 3.5-4.0 kilobases) whose expression is restricted to cells of the granulocytic lineage and parallels the changes in enzymatic activity observed during differentiation. PMID- 3031586 TI - Transcription termination and RNA processing in the 3'-end spacer of mouse ribosomal RNA genes. AB - The 3' termini of ribosomal RNA precursors from mouse FM3A cultured cells are mapped to eight sites within 625 bp downstream from the 3' terminus of 28 S rRNA. Three additional sites are mapped in liver RNA from C3H/He strain mice. Two of them, the sites at 570 bp and 625 bp are assumed to be termination sites in vivo, because they correspond to in vitro termination sites of RNA polymerase I, and 45 S RNAs having these 3' termini decay with kinetics distinct from others. The amount of 45 S RNA having the 3' terminus at other sites is variable among several mouse strains, despite their having the same DNA sequence in these regions. The ability to produce 3' termini in these sites seems to follow Mendel's law of inheritance. Therefore, we postulate that these nine sites are RNA processing sites which are controlled genetically. PMID- 3031587 TI - In vitro molecular genetics as a tool for determining the differential cleavage specificities of the poliovirus 3C proteinase. AB - We describe a completely in vitro system for generating defined poliovirus proteinase mutations and subsequently assaying the phenotypic expression of such mutations. A complete cDNA copy of the entire poliovirus genome has been inserted into a bacteriophage T7 transcription vector. We have introduced proteinase and/or cleavage site mutations into this cDNA. Mutant RNA is transcribed from the altered cDNA template and is subsequently translated in vitro. Employing such a system, we provide direct evidence for the bimolecular cleavage events carried out by the 3C proteinase. We show that specific genetically-altered precursor polypeptides containing authentic Q-G cleavage sites will not act as substrates for 3C either in cis or in trans. We also provide evidence that almost the entire P3 region is required to generate 3C proteinase activity capable of cleaving the P1 precursor to capsid proteins. However, only the 3C portion of P3 is required to generate 3C proteinase activity capable of cleaving P2 and its processing products. PMID- 3031588 TI - Nuclear proteins from lactating mammary glands bind to the promoter of a milk protein gene. AB - The gene for the whey acidic protein (WAP) is expressed specifically in the lactating mammary glands of rodents. We present evidence that nuclear proteins from mammary epithelial cells form a multiple nucleoprotein complex with the WAP gene promoter/upstream region. As monitored by mobility shifts, nuclear proteins from lactating mammary glands and from the mammary cell line MCF-7 form four high affinity complexes with a fragment spanning the region between nucleotides -175 and -88. Nuclear proteins from liver and HeLa cells generate only three high affinity complexes. DNAaseI and ExonucleaseIII protection confirmed the binding of mammary nuclear proteins to specific sequences in the WAP gene upstream region. This is the first report to describe the interaction of nuclear proteins from lactating mammary glands with cognate binding sites in the promoter/upstream region of a milk protein gene. The possibility of the binding sites being candidates for cis-acting regulatory elements governing the regulated expression of the WAP gene is discussed. PMID- 3031589 TI - Nucleotide sequence and characterization of toxR: a gene involved in exotoxin A regulation in Pseudomonas aeruginosa. AB - We have previously reported the discovery and subsequent cloning of a regulatory gene, designated toxR, which appears to regulate the expression of the exotoxin A (ETA) structural gene toxA. Subsequent work by this laboratory has resulted in the subcloning of the toxR gene and its transfer to a high copy number plasmid (pGW28). Functional analysis of the toxR gene using a Tn5 insertion along with toxR deletions indicates that inactivation of toxR results in a dramatic reduction of ETA production. Nucleotide sequence analysis of pGW28 has revealed a 675 bp major open reading frame (225 codons) which could encode for a protein of 24,626 daltons. Using S1 nuclease mapping, the toxR RNA transcript has been shown to originate 20 bp upstream of the presumptive translation initiation codon. Experiments using a toxA specific probe have revealed the the toxR gene product appears to regulate the expression of ETA at the transcriptional level. PMID- 3031590 TI - Nucleotide sequence of the melA gene, coding for alpha-galactosidase in Escherichia coli K-12. AB - Melibiose uptake and hydrolysis in E.coli is performed by the MelB and MelA proteins, respectively. We report the cloning and sequencing of the melA gene. The nucleotide sequence data showed that melA codes for a 450 amino acid long protein with a molecular weight of 50.6 kd. The sequence data also supported the assumption that the mel locus forms an operon with melA in proximal position. A comparison of MelA with alpha-galactosidase proteins from yeast and human origin showed that these proteins have only limited homology, the yeast and human proteins being more related. However, regions common to all three proteins were found indicating sequences that might comprise the active site of alpha galactosidase. PMID- 3031591 TI - Activation of the adenovirus 2 protein IX promoter by DNA replication in a transient expression assay. AB - We have studied the effect of template replication on transcriptional activation of the promoters for the adenoviral protein IX and E1a genes using a short-term transient expression assay. DNA replication mediated modulation of the transcriptional activity of these promoters was monitored using plasmids with limited replication capability conferred by the SV40 minimal origin of DNA replication in the monkey kidney cell line, COS M6. Our results indicate a substantial increase in protein IX promoter directed transcription in a dosage independent manner as a consequence of the DNA replication process. In contrast, the transcriptional activity of the adenovirus 2 E1a promoter was not significantly altered. In addition to this DNA replication-mediated effect, transcription from the protein IX promoter was positively regulated by the adenovirus E1a 13S and E4 gene products but was repressed by the E1a 12S gene product to a limited extent. The evidence suggests that the effect of template replication on the transcriptional activity of plasmid-borne protein IX and E1a gene promoters in this transient expression system closely mimics the situation on the viral chromosome. PMID- 3031594 TI - Conservation of a common 'backbone' in the genetic organization of the IncP plasmids RP4 and R751. PMID- 3031592 TI - The enhancer sequence of human hepatitis B virus can enhance the activity of its surface gene promoter. AB - The transcriptional enhancer sequence has recently been demonstrated in the hepatitis B viral genome. This enhancer sequence has also been shown to increase the activity of HBcAg gene promoter. Using the chloramphenicol acetyltransferase gene expression system, we demonstrated that the intrinsic promoter activity of HBsAg gene promoter was stronger than that of HBcAg gene promoter in both human hepatoma cell lines, Hep3B and HuH-7. Furthermore, we showed that the HBV enhancer sequence not only stimulated the HBcAg gene promoter activity, but also stimulated the HBsAg gene promoter activity in both Hep3B and HuH-7 cells. The enhancer sequence increased the HBsAg gene promoter activity 20-fold in both Hep3B and HuH-7 cell lines, while the increase of the HBcAg gene promoter was 2- and 8-fold in Hep3B and HuH-7 cells, respectively. PMID- 3031595 TI - The human cytomegalovirus immediate early gene promoter is a strong promoter in cultured Drosophila melanogaster cells. PMID- 3031593 TI - Studies on SP6 promoter using a new plasmid vector that allows gene insertion at the transcription initiation site. AB - All the phage-promoter containing subcloning vectors available for in vitro transcription reactions contain a polylinker away from the transcription initiation site. A new SP6 transcription subcloning vector, pCKSP6, has been constructed, in which a gene can be inserted precisely at the transcription initiation site. This was achieved by bringing the BamHI cleavage site into the initiation site. When DNA ends of both insert gene and BamHI cleaved pCKSP6 are made blunt-ended using a single strand specific nuclease, the in vitro transcripts of the recombinant DNA by SP6 RNA polymerase will contain only the gene sequence immediately after the initiation base G. Mung bean nuclease was used to generate a series of mutants resulting from step-wise deletion of single base pairs around the initiation site. Transcription assays with these SP6 promoter mutants revealed that not only the sequence immediately upstream of the initiation site but also the six base pairs from position +1 to +6 are important elements for promoter binding and/or transcription initiation activity. Furthermore, there appears to be a hierarchy of importance of each base pair in the order of position +1 greater than +2 greater than +3 greater than +4, +5, +6, -1, -2. PMID- 3031596 TI - Nucleotide sequence of the long terminal repeat of the avian retrovirus RAV-1: evolution of avian retroviruses. PMID- 3031597 TI - Properties of a U1 RNA enhancer-like sequence. AB - The properties of a X.laevis U1B snRNA gene enhancer have been studied by microinjection in Xenopus oocytes. The enhancer-like sequence, defined as a short DNA stretch that is able to activate transcription in an orientation independent manner, is interchangeable between different U snRNA genes. The enhancer sequence alone does not, however, efficiently activate transcription from an SV40 pol II promoter but regains its activity when combined with the U-gene specific proximal sequence element. DNase I protection experiments show that the X.laevis U1B enhancer can interact specifically with a nuclear factor present in mammalian cells. PMID- 3031598 TI - The sequence motifs that are involved in SV40 enhancer function also control SV40 late promoter activity. AB - The simian virus 40 (SV40) enhancer element is constituted of two domains which contain sequences important for late transcription (M. Ernoult-Lange, F. Omilli, D. O'Reilly and E. May, J. Virol. 61, 167-176, 1987). By analysing a series of clustered point mutations generated throughout the enhancer region we mapped domain I from nt 232 to 272 and domain II from nt 184 to 216. These two domains which are required for late promoter activity both in the presence and in the absence of T antigen correspond closely to the domains B and A respectively, identified for enhancer function (M. Zenke, T. Grundstrom, H. Matthes, M. Wintzerith, C. Schatz, A. Wildeman and P. Chambon, EMBO J., 5, 387-397, 1986). Similarly to the enhancer function the late promoter elements defined by these two domains contain multiple sequence motifs. Moreover there is a striking overlap between the sequence motifs within domain A, active for early enhancer function and those within domain II involved in efficient late transcription. PMID- 3031599 TI - Xenopus tropicalis U6 snRNA genes transcribed by Pol III contain the upstream promoter elements used by Pol II dependent U snRNA genes. AB - We have cloned and sequenced a 977bp DNA fragment, pXTU6-2, that represents the transcription unit for a Xenopus tropicalis U6 RNA gene. This basic repeating unit is reiterated ca.500-fold per haploid genome. Oocyte injections of pXTU6-2 led to the transcription of a mature-sized U6 RNA that, however, lacked internal 2'-O-methylations. These posttranscriptional modifications of U6 RNA might be cytoplasmic and could require its association with U4 RNA to be accomplished. The low alpha- amanitin sensitivity of U6 RNA synthesis in oocytes suggested that U6 RNA is transcribed by RNA polymerase III, consistent with features of the U6 RNA molecule which also contains a Box A- like intragenic control region. Inspection of X. tropicalis, mouse and human U6 DNA upstream sequences revealed the presence of a TATA box as well as of the proximal and enhancer (octamer motif) elements contained in snRNA genes transcribed by RNA polymerase II. We propose that U6 RNAs are synthesized by a specialized transcription complex consisting of RNA polymerase III and transcription factors, some of which are very likely shared with RNA polymerase II promoters. PMID- 3031600 TI - Transcripts within the replication origin, oriC, of Escherichia coli. AB - Transcription start and termination sites were mapped in the E. coli replication origin, oriC. Outward transcription from within oriC (promoters Pori-r and Pori l) was found to start in vivo at position 178 for Pori-l and at positions 294 and 304 for Pori-r, respectively. These transcripts were terminated after 100-150 bases, at terminators designated Tori-l and Tori-r. Transcription from the 16 kd promoter, which lies clockwise adjacent to oriC and promotes transcription toward oriC, started at position 757 and gave transcripts with 3' ends at several positions within and to the left of the minimal replication origin. However, the majority of transcripts traversed the whole oriC region, and were not terminated within the DNA segment tested. Transcription of the chromosomal 16 kd gene was negatively regulated by DnaA protein and positively affected by dam methylation. The possible function of these transcripts is discussed. PMID- 3031602 TI - Nucleotide sequence analysis of alpha-amylase and thiol protease genes that are hormonally regulated in barley aleurone cells. AB - We have determined the nucleotide sequences of Amy32b, a type A alpha-amylase gene, and of the gene for aleurain, a thiol protease closely related to mammalian cathepsin H. Both are expressed in barley aleurone cells under control of the plant hormones gibberellic acid and abscisic acid, but only aleurain is expressed at high levels in other barley tissues. Sequence analysis indicates that the 5' end of the aleurain gene, comprising 3 exons and 2 introns, may have become associated with the remainder of the gene, encoding the protease domain of the protein, by some sort of recombination event. This 5' domain of the gene is very G + C-rich and is flanked by inverted repetitive sequences. We found two different groups of homologous sequence elements. The first group consists of four blocks of sequences conserved in the same spatial arrangement in both genes; these are arranged at similar intervals upstream from the Amy32b TATA box and from a TATA box present in intron 3 of aleurain, outside of the 5' domain and upstream from the protease domain. A part of two of these conserved sequences is similar to the core sequence of certain enhancer elements characterized from mammalian cells. The second group of homologous elements is present in the upstream region of both genes. We speculate that these conserved sets of sequences may have some role in either the tissue specificity of expression of the genes or in some part of the hormonal regulation imposed on them. PMID- 3031601 TI - Molecular cloning and characterization of rat estrogen receptor cDNA. AB - A cDNA clone of rat uterus estrogen receptor (ER) has been isolated and sequenced. This clone contains a complete open reading frame encoding 600 amino acid residues which is 5 and 11 amino acids larger than the corresponding molecules of human and chicken, respectively. The molecular weight of this protein is calculated to be 67,029. When this clone was ligated to the pSV2 vector and transfected into COS7 cells, a protein was produced that had the same affinity to estrogen as rat uterus ER. This sequence shows 88% homology with human ER; 528 amino acids are identical and 14 amino acids are conservative substitutions. The comparison of rat, human and chicken ER sequences indicate the presence of three highly conserved regions suggesting that these regions play important roles in ER function. The putative DNA-binding domain is completely identical in rat, human and chicken. The C-terminal half region which is thought to be the estrogen binding domain is also highly conserved and is rich in hydrophobic amino acid residues. Southern blot analysis of genomic DNA with ER cDNA as a probe has shown that related sequences are present in the genome. PMID- 3031603 TI - Characterization of a human 'midisatellite' sequence. AB - We have examined the structure and DNA sequence of a human genomic locus that consists of a large hypervariable region made up of repeats of a simple sequence. With several restriction enzymes, the locus shows many restriction fragments that vary quantitatively as well as qualitatively. Other restriction enzymes produce only a single, high-molecular-weight fragment at this locus. Almost all of the fragments are revealed with a simple sequence probe. Southern transfers of the high-molecular-weight restriction fragments produced by the restriction enzymes NotI and SfiI, resolved by pulsed-field gel electrophoresis, gave at most two fragments, demonstrated to be allelic, showing that the majority of the restriction fragments seen in the complex patterns are at a single locus. The estimated size of the region homologous to the probe varied from 250 to 500 kilobases. DNA sequencing indicated that the region consists of tandem repeats of a 40-base-pair sequence. Some homology was detected to the tandem repeating units of the insulin gene and the zetaglobin pseudogene hypervariable regions, and to the "minisatellite" DNA at the myoglobin locus. PMID- 3031604 TI - Aggregation of DNA by analogs of spermidine; enzymatic and structural studies. AB - A homologous series of spermidine analogs, with defined abilities to replace the natural polyamine in supporting cell growth, was examined for its influence on the structure of supercoiled, aggregated DNA and on the ability of the DNA aggregates to act as substrates for various enzymes. The concentration of amine necessary to aggregate negatively supercoiled Col E1 DNA was progressively increased as the diaminobutane moiety of spermidine was extended beyond 5 methylene groups. 1H- and 31P-NMR spectroscopy suggested that less rigid DNA aggregates were formed by spermidine analogs than by spermidine itself. Spermidine and its analogs differentially modulated the activities of bacterial and mammalian type I topoisomerases and EcoRI restriction endonuclease on aggregated DNA in a manner reminiscent of the abilities of the amines to stimulate cell growth. When DNA was not aggregated, the influence of the various amines on these reactions was almost identical. These results are discussed in relation to the structures of the DNA aggregates in the presence of the various triamines. PMID- 3031605 TI - The length but not the sequence of the polyoma virus late leader exon is important for both late RNA splicing and stability. AB - Polyoma virus late RNA processing provides a convenient model system in which to study the mechanics of splicing in vivo. In order to understand further the role of the untranslated "late leader" unit in late RNA processing we have constructed a group of polyoma viruses with deletions and substitutions in the leader exon. This has allowed us to determine that there is a minimum exon size required for both pre-mRNA splicing and stability in this system. We show here that the non viability of a mutant (ALM) with a 9 base late leader unit is due to a general defect in late RNA splicing. In addition, ALM-infected cells show at least 40 fold depression in the accumulation of late nuclear RNA (spliced or unspliced). The ALM late promoter, however, functions nearly normally. Substituted leader variants with 51- to 96-base long exons of unrelated sequence are viable (G. Adami and G. Carmichael, J. Virol. 58, 417-425, 1986). We show here that late RNA from one of these substituted leader mutants (containing a 51-base leader exon) is spliced at wild type levels, with virtually no defect in accumulation. Thus, in the polyoma system, splice sites separated by only 9 bases can inhibit each others usage, presumably by steric interference. We suggest that this type of inhibition leads to extreme RNA instability. PMID- 3031606 TI - Nucleotide sequence and expression of the cloned gene of bacteriophage SP6 RNA polymerase. AB - The coding region of the gene for bacteriophage SP6 RNA polymerase was cloned into pBR322, and its entire nucleotide sequence was deduced. The predicted amino acid sequence for the polymerase consists of 874 amino acid residues with a total molecular weight of 98,561 daltons. Comparison of the amino acid sequence with that of T7 RNA polymerase reveals that regions with partial homology are present along the sequence. The coding region of SP6 RNA polymerase was inserted into an E. coli expression vector. The polymerase gene was efficiently expressed in E. coli cells, and the enzymatic properties of the expressed polymerase were very similar to those of the enzyme synthesized in SP6 phage-infected Salmonella typhimurium cells. PMID- 3031607 TI - Activity of a Streptomyces transcriptional terminator in Escherichia coli. AB - A 205bp DNA fragment from the Streptomyces multi-copy plasmid pIJ101 has in vivo terminator activity both in Streptomyces lividans and in Escherichia coli. Termination of RNA synthesis, detected by high-resolution S1 nuclease mapping, occurs at precisely the same nucleotides in both organisms. This suggests that the E. coli RNA polymerase recognizes the same sequence elements and chooses the point(s) of termination in the same way as the corresponding S. lividans enzyme. PMID- 3031608 TI - Charons 36 to 40: multi enzyme, high capacity, recombination deficient replacement vectors with polylinkers and polystuffers. AB - New phage lambda based cloning vectors, Charons 36-40, have been constructed which allow cloning of large (up to 24 kb) DNA fragments with up to sixteen cloning enzymes. Several of these could not be used previously with lambda vectors. Clones produced with these vectors can be propagated under recombination deficient conditions. A novel polystuffer method has been developed that permits vector arms to be purified by simple precipitation and which allows reliable identification of clones that have reincorporated any part of the stuffer. Three of the vectors are available with amber mutations in essential genes. PMID- 3031609 TI - Identification of an Epstein-Barr virus-specific desoxyribonuclease gene using complementary DNA. AB - We have recently obtained 18 distinct cDNA clones representing different genes expressed in the early phase of EBV infection. One of them, c37, which is situated at the position 12907-122451 in the B95-8 viral genome, is shown here to code for a viral desoxyribonuclease [DNase]. Cell free translation of c37 selected messenger RNA yielded a protein of about 52 KDa which was immunoprecipitated by a high EA titer serum from nasopharyngeal carcinoma patient. This protein showed a DNase activity which was resistant to high salt concentrations (150 to 300 mM KCl) and was specifically neutralized by EA positive serum. These properties are typical of the EBV-specific DNase activity that we recently described in chemically induced EBV-transformed lymphoid cells. The same results were obtained on cell-free translation of the native RNA synthesized in vitro from pGEM-37 plasmid containing the entire c37 cDNA sequence (1.53 Kb). These data indicate that the BGLF5 open reading frame contained in c37 encodes for the EBV-specific DNase. PMID- 3031611 TI - Hybrid pUC vectors for addition of new restriction enzyme sites to the ends of DNA fragments. PMID- 3031610 TI - A cellular protein binds to a conserved sequence in the adenovirus type 2 enhancer. AB - A sensitive gel retention assay has been utilized to detect proteins from uninfected Hela nuclei which interact with the adenovirus type 2 enhancer. This assay has been employed to monitor fractionation of nuclear extracts. Three enhancer binding factors were resolved by chromatography on DEAE-Sepharose and one of the factors was further purified by chromatography on heparin-Sepharose. DNase protection experiments have shown that the heparin-Sepharose fraction contains a factor which binds predominantly to the conserved sequence GTGGAAATTT present at position 160 in the adenovirus type 2 genome and found in many viral and cellular enhancers. Protection of this sequence from DNase I digestion was abolished by competition with a synthetic duplex oligonucleotide spanning bases 144-181. This region corresponds to the sequence defined by Hen et al. as possessing enhancer function. Competition experiments indicated that the enhancer binding factor also bound, albeit with reduced affinity, to multiple sites in the Ela upstream region located between positions 192 and 353. Within the sequences which compete are regions with homology to the high affinity site at position 160. The enhancer binding factor also binds with high affinity to sequences within the SV40 enhancer demonstrating that this factor interacts with sequences common to both the adenovirus and SV40 enhancers. PMID- 3031613 TI - Unusual structure, evolutionary conservation of non-coding sequences and numerous pseudogenes characterize the human H3.3 histone multigene family. AB - The genomic organization of the replication-independent, basally expressed, human H3.3 gene is atypical of traditional histone gene organization. The gene contains 3 introns totalling 7.8 kb and unusual direct repeats flank all three intron-exon splice junctions. The transcription initiation site was mapped by S1 nuclease protection analysis and confirms that cDNA clones previously reported were full length. Sequence similarities between regions at the 5' and 3' termini of this human gene and a chicken H3.3 gene lead us to propose that either the previous assignments of termini of the chicken gene are in error, or there are alternative transcription start and polyadenylation sites. The 85% base matching of human and chicken H3.3 3'UTR sequences for 520 bases is unprecedented among homolog 3'UTR segments, especially considering that these species are separated by over 250 Myr of evolution. We also present the sequence of three related processed human H3.3 pseudogenes and provide evidence demonstrating that most of the 20 to 30 copies of the H3.3 gene within the human genome are in fact processed pseudogenes. PMID- 3031612 TI - Nuclear factors binding to the human immunoglobulin heavy-chain gene enhancer. AB - The human immunoglobulin heavy chain (IgH) gene contains at least two tissue specific regulatory regions, which are similar to the mouse IgH gene. One is the J-C enhancer and another is located in the 5' promoter region. Using an electrophoretic mobility shift assay and DNase I footprint, we have examined the interaction of factors in B cell nuclear extracts with the two regulatory regions of the human IgH gene. We have identified a nuclear factor in mouse B cell nuclear extracts which bound to specific sequence in the human IgH enhancer. This factor is apparently not present in mouse fibroblast nuclear extracts. We also found factor(s) which bound to the highly conserved octanucleotide sequence within the human IgH enhancer and 5' promoter regions. PMID- 3031615 TI - Nucleotide sequence of the bovine parainfluenza 3 virus genome: the genes of the F and HN glycoproteins. AB - By analysing complementary DNA clones constructed from genomic RNA of bovine parainfluenza 3 virus (BPIV3), we determined the nucleotide sequence of the region containing the entire F and HN genes. Their deduced amino acid sequences showed about 80% homologies with those of human parainfluenza 3 virus (HPIV3), about 45% with those of Sendai virus, and about 20% with those of SV5 and Newcastle disease virus (NDV), indicating, together with the results described in the preceding paper on the NP, P, C and M proteins of BPIV3, that BPIV3, HPIV3 and Sendai virus constitute a paramyxovirus subgroup, and that BPIV3 and HPIV3 are very closely related. The F and HN proteins of all these viruses, including SV5 and NDV, however, were shown to have protein-specific structures as well as short but well-conserved amino acid sequences, suggesting that these structures and sequences are related to the activities of these glycoproteins. PMID- 3031614 TI - Nucleotide sequence of the bovine parainfluenza 3 virus genome: its 3' end and the genes of NP, P, C and M proteins. AB - We present the nucleotide sequence of bovine parainfluenza 3 virus (BPIV3) genome from its 3' end to the opening region of the F gene, through the NP, P plus C, and M genes. Comparison of the sequence with those reported for other paramyxoviruses indicated that BPIV3 was most similar to human parainfluenza 3 virus (HPIV3), and also very similar to Sendai virus in the structural make-up of its genome and the amino acid sequences of its gene products, suggesting that these three viruses constitute a paramyxovirus subgroup from which Newcastle disease and measles viruses are separable. In BPIV3 and Sendai virus, the NP and M proteins, the main structural elements, were more highly conserved than the functionally important P and C proteins. This tendency was also observed even in BPIV3 and HPIV3. Virus-specific amino acid sequences of the NP and M proteins were found at the carboxyl and amino terminal regions, respectively. BPIV3 M mRNA was found to have aberrations in its poly A attachment site. PMID- 3031616 TI - Reconstruction of human alpha thalassemia-2 genotypes in monkey cells. AB - The human adult alpha globin genes, alpha 2 and alpha 1, are contained within two tandemly arranged duplication units. Each unit spans 4 kb of DNA, and contains three homology blocks (X, Y, Z) separated by non-homologous sequences. Segmental DNA recombination processes between the two units have resulted in high frequencies of two types of deletions in certain human populations, each deletion removing one alpha globin gene from chromosome 16, (alpha-thalassemia 2). In order to study the molecular mechanisms of alpha-thalassemia 2, and of homologous DNA recombination in general in mammalian cells, we have reconstructed these two alpha-thalassemia 2 genotypes in monkey cells. The two duplication units have been cloned in an SV40 origin-containing vector, and transfected into COS 7 cells. Newly replicated plasmid DNA was isolated and analyzed by Southern blot hybridization. Homologous DNA recombination occurs with high frequencies (10-20% per kb of homology), and this generates both types of alpha-thalassemia 2 deletions on the episomes in the monkey cells. Removal of the 5' end of either one, or both, of the X blocks prior to DNA transfection affects the relative frequencies of the two alpha-thalassemia 2 genotypes in a novel way. We consider and discuss these results in terms of several alternative models. Our data suggest the existence of hot spot(s) for initiation of homologous DNA recombination, or recombination promoting element(s), in a specific region of the human adult alpha globin locus. A DNA sequence that defines the boundaries of the two duplication units, and has been implicated in the initiation of gene conversion of the two X blocks, is contained within this region. PMID- 3031617 TI - Nonrandom distribution of MMTV proviral sequences in the mouse genome. AB - Integrated sequences of mouse mammary tumor virus (MMTV) have been localized in the genomes of five inbred mouse strains (Balb/c, C3H, DBA/2, A.TH, 129-SV) and one mammary tumor cell line (GR). Two major classes of MMTV sequences have been detected in mouse DNA fractions as obtained by Cs2SO4/BAMD (3,6-bis (acetatomercurimethyl)dioxane) density gradient centrifugation. The first one corresponds to previously described endogenous sequences (Mtv loci), whereas the second one corresponds to endogenous sequences not previously known, and/or recently acquired; in the case of GR cells exogenous sequences may also be present in this class. The genome distribution is somewhat different for the two classes of sequences, the first one being practically only present in the lightest DNA segments of the mouse genome (GC congruent to 38%); the second one being also represented in heavier segments (GC congruent to 43%). This integration pattern suggests that "ancient" endogenous sequences are practically only localized in genome segments of roughly matching composition, whereas exogenous and recently acquired endogenous MMTV sequences may also be present in heavier fractions. PMID- 3031618 TI - Minor-groove-modified oligonucleotides: synthesis of decadeoxynucleotides containing hypoxanthine, N2-methylguanine and 3-deazaadenine, and their interactions with restriction endonucleases Bgl II, Sau, 3AI, and Mbo I (Nucleosides and Nucleotides Part 75). AB - Decadeoxynucleotides containing hypoxanthine, N2-methylguanine, 3-deazaadenine in the recognition sequences of restriction endonucleases Bgl II, Sau 3AI, and Mbo I were synthesized. These decanucleotides modified in the base moieties facing in to the minor groove were strongly resistant to hydrolysis by Bgl II and partially resistant to that of Sau 3AI and Mbo I. The decadeoxynucleotide containing 3 deazaadenine in place of adenine was bound to Bgl II strongly, whereas the nucleotides containing hypoxanthine and N2-methylguanine were bound less tightly. PMID- 3031619 TI - Nucleotide sequence of the Escherichia coli mutH gene. AB - The complete nucleotide sequence of mutH gene from E. coli has been determined. Based on the deduced amino acid sequence, the MutH protein has a molecular weight of 25.4 kdaltons in agreement with the previous estimates based on SDS polyacrylamide gel electrophoresis of the purified protein. Deletion analysis of the DNA sequences upstream of mutH has identified the promoter region for this gene. Two independently isolated temperature sensitive alleles of the mutH gene have also been sequenced. One mutation results in an amino acid change at position 27 (thr to leu) while the other occurs at position 156 (asp to asn). PMID- 3031621 TI - Determination of vector: insert junctions in lambda gt10 cDNAs that do not recut with EcoRI. Nucleotide sequence of the lambda imm434 HindIII-EcoRI DNA fragment encoding part of the cI protein. PMID- 3031620 TI - Regulation of the herpes simplex virus type 1 late (gamma 2) glycoprotein C gene: sequences between base pairs -34 to +29 control transient expression and responsiveness to transactivation by the products of the immediate early (alpha) 4 and 0 genes. AB - The glycoprotein C (gC) gene of herpes simplex virus type 1 is a true late gene, in that its expression occurs late in infection with a strict requirement for viral DNA replication. Recently, we reported on gC expression during infection with mutant viruses carrying deletions in the gC gene promoter. Analysis of RNA extracted from cells infected with individual mutants showed that the DNA sequences required for regulated expression of this late gene lie within bases 34 to +124 relative to the 5' end of the mRNA. In the present study, the deleted gC promoter sequences were fused to the bacterial chlorampheniol acetyltransferase (CAT) gene and expression was measured in short-term transfection assays after transactivation by infection with HSV or cotransfection with a second plasmid carrying the immediate early genes of HSV-1. The 63 base pair sequence located between -34 to +29 on the gC promoter was sufficient to give induction of CAT activity following infection and on cotransfection with plasmids which code for the immediate early gene products ICP4 and ICPO. This 63 base pair region contains the TATA homology and the transcriptional start site of the gC gene, and apparently contains specific promoter elements not found in a similar region of the HSV TK promoter. This was shown by removing the distal upstream region of the TK promoter, 5' to -37, and found that the TK gene was no longer activated by infection or cotransfection with an alpha 4-alpha 0 gene containing plasmid. PMID- 3031622 TI - Nucleotide sequence of the human parainfluenza virus 3 matrix protein gene. PMID- 3031623 TI - Architecture of models for prebiotic synthesis of nucleic acids: template directed synthesis of oligoguanylates using N-cyanoimidazole. AB - N-Cyanoimidazole is an efficient condensing agent for the polymerization of guanosine 5'-phosphate (pG) on a poly(C) template in an aqueous solution. At 0 degree C, up to about 30% of input pG was converted to a mixture of oligomers with a mean chain length of up to 7. The effect of divalent metal ions in the polymerization of pG on a poly(C) template was not so considerable as in that of oligo(A) on a poly(U) template. In the polymerization of pG, the moderate yields were obtaind in the presence of Co2+, Ni2+ and Cu2+. PMID- 3031624 TI - The expression of human tumor necrosis factor in E. coli. AB - cDNA of human natural TNF (n-TNF) obtained by stimulating human leukemic B cell line (Ball-1) with Sendai virus was cloned. Valine-started-TNF (V-TNF) gene was constructed from the cDNA and expressed in E.coli HB101 under the control of a trp promoter by the induction of 3-indoleacrylic acid. The expression level of V TNF clone was about 10% of the total E.coli protein. On the other hand, the expression level of glutamine started-TNF (Q-TNF) gene having the same SD-ATG sequence which was constructed from V-TNF gene was as low as about 1/20 of that of V-TNF. The nucleotide sequence around ATG (-4 approximately +12) of Q-TNF gene was randomly changed without modifying the coded amino acid sequence, resulting to obtain high expression clones as similar TNF protein yield as that of V-TNF. These clones possessed A residue rich sequence around the initiation codon ATG. These results show that some correlation might exist between the high expression rate and A residue rich sequence around the initiation codon. PMID- 3031625 TI - Products distribution in the oxidation of thymine with hydroxyl radicals. AB - In the oxidation of thymine with hydroxyl radical generated from L-ascorbic acid/copper(II) ion/O system, four reaction products, thymine glycol(TG), N formyl-N'-pyruvylurea(FPU), 5-hydroxymethyluracil(HMU) and 5-hydroxy-5-methyl barbituric acid(HMBA) were obtained. A reaction scheme was proposed to explain the products distribution observed. PMID- 3031626 TI - Total assignments of the 1H NMR of decanucleotide duplexes containing the recognition sites of restriction enzymes. AB - Complementary DNAs, decanucleotides, were synthesized by solid phase synthesis. They contain six base pairs with different sequences in the center, flanked by GC pairs to stabilize the ends of the helix. All the imino and non-exchangeable protons of these decamers in aqueous solution were assigned by NOESY experiment. Sequence dependent exchange rates of imino protons and the chemical shifts of non exchangeable protons were examined. PMID- 3031627 TI - Studies on griseolic acid derivatives. III. Synthesis and biological activities of the adducts of griseolic acid and their base exchanged derivatives. AB - Addition reactions across the double bond of griseolic acid were investigated. Dihydrogriseolic acid was obtained by a reduction of the adduct having halogen at 4' position. The ring juncture of the two five membered rings of the dihydro derivatives was all "cis" configuration. An acetolysis of the protected dihydro derivative gave corresponding 1'-acetoxy sugar. A glycosidation of this sugar derivative with silylated bases gave base exchanged derivatives of the dihydrogriseolic acid. The influence of the base moiety and the double bond to the PDE inhibitory activity was investigated. As a result, we found that this type of compounds had a weaker inhibitory activity than the corresponding compounds which had an original double bond or a dihydro bond that made the ring juncture of the two five membered ring "trans". PMID- 3031628 TI - Biologically active analogs of thymopentin with enhanced enzymatic stability. AB - Thymopentin, a synthetic pentapeptide fragment of thymopoietin (residues 32-36, Arg-Lys-Asp-Val-Tyr) is biologically active but susceptible to proteolytic digestion. Analogs were synthesized and studied for biological activity and susceptibility to peptidases. Amino acid changes were incorporated at positions known to not affect activity adversely and N-terminal acetylation and C-terminal amidation were used to increase resistance to proteolytic degradation by exopeptidases. Ac-Pro2-TP5-NH2 and Aib2-TP5-NH2 retained activity and were shown to exhibit a high degree of stability when incubated in human serum. PMID- 3031629 TI - Pharmacologic characterization and autoradiographic distribution of binding sites for iodinated tachykinins in the rat central nervous system. AB - P-type, E-type, and K-type tachykinin binding sites have been identified in the mammalian CNS. These sites may be tachykinin receptors for which the mammalian neuropeptides substance P, neuromedin K, and substance K are the preferred natural agonists, respectively. In the present investigation, we have compared the pharmacology and the autoradiographic distribution of CNS binding sites for the iodinated (125I-Bolton-Hunter reagent) tachykinins substance P, eledoisin, neuromedin K, and substance K. Iodinated eledoisin and neuromedin K exhibited an E-type binding pattern in cortical membranes. Iodinated eledoisin, neuromedin K, and substance K each labeled sites that had a similar distribution but one that was considerably different from that of sites labeled by iodinated substance P. CNS regions where there were detectable densities of binding sites for iodinated eledoisin, neuromedin K, and substance K and few or no sites for iodinated substance P included cortical layers IV-VI, mediolateral septum, supraoptic and paraventricular nuclei, interpeduncular nucleus, ventral tegmental area, and substantia nigra pars compacta. Binding sites for SP were generally more widespread in the CNS. CNS regions where there was a substantial density of binding sites for iodinated substance P and few or no sites for iodinated eledoisin, neuromedin K, and substance K included cortical layers I and II, olfactory tubercle, nucleus accumbens, caudate-putamen, globus pallidus, medial and lateral septum, endopiriform nucleus, rostral thalamus, medial and lateral preoptic nuclei, arcuate nucleus, dorsal raphe nucleus, dorsal parabrachial nucleus, parabigeminal nucleus, cerebellum, inferior olive, nucleus ambiguus, retrofacial and reticular nuclei, and spinal cord autonomic and somatic motor nuclei. In the brainstem, iodinated substance P labeled sites in both sensory and motor nuclei whereas iodinated eledoisin, neuromedin K, and substance K labeled primarily sensory nuclei. Our results are consistent with either of two alternatives: (1) that iodinated eledoisin, neuromedin K, and substance K bind to the same receptor site in the rat CNS, or (2) that they bind to multiple types of receptor sites with very similar distribution. PMID- 3031630 TI - Possible recognition of the GnRH receptor by an antiserum against a peptide encoded by nucleotide sequence complementary to mRNA of a GnRH precursor peptide. AB - Two peptides, rHRnG and hproHRnG, which were encoded by the nucleotide sequences complementary to mRNA of rat hypothalamic gonadotropin-releasing hormone (GnRH) and human placental proGnRH(-3-13), respectively, were synthesized. A remarkable hydropathic anti-complementarity was observed in the N-terminal region between hproHRnG and human proGnRH(-3-13). Neither hproHRnG nor rHRnG bound GnRH in ELISA unless extremely high concentrations of peptides were used. 125I-GnRH failed to bind with either rHRnG or hproHRnG previously coated polypropylene tubes. Antisera against these peptides were generated in rabbits. All the rabbits produced antibodies with high titer as tested by ELISA. One rabbit immunized with hproHRnG showed markedly reduced serum testosterone levels as compared with those of other rabbits. Intravenous administration of 1 ml serum from this rabbit, antiserum R281, into ovariectomized rats significantly decreased plasma LH. Using antiserum R281, about 10% of female rat pituitary cells were stained by immunohistochemistry. The staining was specific to hproHRnG since it was abolished by preabsorption of the antiserum with hproHRnG, but not with rHRnG, GnRH, LH nor any other peptide tested. This particular antiserum may have recognized the GnRH receptor, and thereby interfered with the action of endogenous GnRH. These results appear to be in agreement with the view that there is a structural similarity between the receptor for a peptide and the so-called complementary peptide. PMID- 3031632 TI - Synthetic peptides from C-reactive protein containing tuftsin-related sequences. AB - Peptides containing Lys-Pro-Arg or Thr-Lys-Arg segments corresponding to various regions of human C-reactive protein were synthesized. The peptides prepared were composed of amino acid residues, 37-58, 51-58, 173-187 and 181-187 of C-reactive protein. The relationship between C-reactive protein, its synthetic fragments and tuftsin (Thr-Lys-Pro-Arg) was investigated in binding studies, enhancement of phagocytosis and change in cyclic nucleotide levels of mouse macrophages. The peptides AA 51-58 and 181-187 did enhance macrophage phagocytosis capacity to a similar extent to that of tuftsin. They showed however only negligible binding to the cells. The effect of C-reactive protein and the synthetic peptides on metabolic activity of neutrophils was also investigated. It was shown that the peptides inhibited to some degree superoxide production, lysozyme release and Vitamin B12 binding protein release from neutrophils in the absence and presence of the stimulants, PMA or Con A. Comparable activity with tuftsin was not found. PMID- 3031633 TI - [Role of genetic factors in the etiology of cancer of the large intestine]. PMID- 3031631 TI - Comparison of the effects of CRF and stress on levels of pituitary cyclic AMP and plasma ACTH in vivo. AB - We have previously reported that acute stress increases levels of rat pituitary cyclic AMP in vivo. The present study was conducted to test the hypothesis that stress-induced increases in pituitary cyclic AMP in vivo were mediated via CRF. We compared the effects of various stressors with the effects of CRF or epinephrine administration on pituitary cyclic AMP and plasma ACTH responses in vivo. Stressors, epinephrine or CRF increased levels of pituitary cyclic AMP. Pituitary cyclic AMP response to either immobilization or CRF was much greater at light onset than at lights off in rats maintained on a 12 hr light:12 hr dark lighting regimen. In rats with pituitary stalk transections, footshock did not increase levels of pituitary cyclic AMP, suggesting that some factor of central origin was required for this stress response. Exogenous CRF administration did increase levels of pituitary cyclic AMP in stalk-transected rats, while epinephrine increased levels in sham-operated but not in stalk-transected rats. Antisera to CRF markedly decreased pituitary cyclic AMP response to exogenous CRF administered 6 min following antisera and partially attenuated pituitary cyclic AMP response to forced running. Taken as a whole these data support a major role for CRF in the pituitary cyclic AMP response to stress. PMID- 3031634 TI - Breast lesions diagnosed by fine needle aspiration. AB - Fine needle aspiration (FNA) of the breast offers patient and clinican a rapid, non-morbid, inexpensive, and highly accurate means of diagnosing both benign and malignant breast lesions. Breast FNA can replace frozen section diagnosis, and in fact has the advantage of providing a known diagnosis before the time that operations are performed. This situation enables the patient and surgeon to discuss and plan therapeutic alternatives in a rational atmosphere. As with any other technique, experience of the aspirator and diagnostician minimizes false positive and false negative rates. PMID- 3031635 TI - Fibrohistiocytic tumors of soft tissues. An immunohistochemical study of 183 cases. AB - 183 cases of soft tissue tumors were studied utilizing the immunoperoxidase technique to demonstrate alpha-1-antitrypsin, ferritin and lysozyme. The series comprises 50 malignant lesions, 34 intermediate malignancy lesions, 99 benign lesions of fibrohistiocytic origin, and 23 malignant tumors of non fibrohistiocytic origin. The actual results of the study are as follows: alpha-1 antitrypsin, ferritin and lysozyme were always absent in 10 fibrosarcomas, 2 liposarcomas, 2 Ewing sarcomas, 3 synovial sarcomas, 4 neurofibrosarcomas, and 2 rhabdomyosarcomas, but in 24 malignant fibrous histiocytomas, 34 cases of dermatofibrosarcoma protuberans and 102 benign fibrohistiocytic lesions, these activities were present in a percentage that ranged between 12% and 38% (average 25%). Differences in the frequency of positive reactions did not occur between benign and malignant fibrohistiocytic lesions. The immunohistological examinations carried out have, therefore, only a very limited value for the practical diagnostic evaluation, but, when positive, are important to clarify the histogenesis of the tumor. PMID- 3031636 TI - The histopathologic identification of CMV infected cells in biopsies of human renal allografts. An evaluation of 100 transplant biopsies by in situ hybridization. AB - In order to determine the incidence and significance of CMV infected cells within human renal allograft biopsies 100 transplant biopsies were examined for the presence of CMV DNA within the renal tissue specimens using the in situ hybridization technique. In 41 cases CMV infected cells were predominantly found within proximal tubular epithelial cells, although typical nuclear inclusion ("owl eyes") were absent. In only one case was CMV detected within a few glomerular cells. The presence of CMV infected cells within allograft biopsies does not correlate with active CMV infection of the patients at the time of biopsy. There are no significant differences in the distribution of primary and secondary CMV infections between patients with positive and negative biopsy findings. No significant differences as to the histological alterations between CMV infected and non-infected biopsies could be found. The data give evidence that the renal allograft is more often affected by CMV than is generally appreciated. The in situ hybridization technique may be useful for the fast detection of latently CMV infected cells in renal transplants and thus may influence the choice of therapeutic steps early after transplantation. Furthermore, it may facilitate the diagnosis of interstitial nephritis due to virus infection if typical nuclear inclusions in routinely stained tissue sections are absent. PMID- 3031637 TI - Parvovirus infections in childhood. AB - Human parvovirus (HPV) infections have been associated with several clinical syndromes. The virus is the etiologic agent of erythema infectiosum (fifth disease) and the primary cause of aplastic crises in children with sickle cell disease and other hemolytic anemias. Some individuals with an acute symmetric polyarthritis were found to have a recent HPV infection. Although two stillborn infants with proved HPV infections have been identified, the exact relationship among HPV, intrauterine infection, and subsequent fetal damage requires further research. PMID- 3031638 TI - [Pain and endogenous opioid peptides]. PMID- 3031640 TI - Morphine-6-glucuronide has high affinity for the opioid receptor. PMID- 3031639 TI - Effects of lithium on calmodulin-stimulated adenylate cyclase activity in cortical membranes from rat brain. AB - The influence of lithium on calmodulin-stimulated adenylate cyclase activity has been studied in vitro and after chronic treatment. Chronic lithium treatment decreased calcium-calmodulin-stimulated adenylate cyclase activity in rat cortical membranes, while no effect was observed on GTP-stimulated activity. Lithium in vitro inhibited adenylate cyclase activity stimulated by isoprenaline, GTP or calcium-calmodulin. Calcium-calmodulin-stimulated activity was more sensitive to lithium (2 mM) than isoprenaline- and GTP-stimulated activities (5 mM) and activities by these agents combined. Lithium had no effect on the unstimulated enzyme activity. The inhibitory effect of lithium in vitro on calcium-calmodulin-stimulated adenylate cyclase activity was antagonized by magnesium. The inhibition induced by lithium in vitro on the GTP-stimulated adenylate cyclase activity was increased by substituting manganese for magnesium in the assay media. Furthermore, the manganese-stimulated activity was also reduced by lithium. The latter effect was not observed in calmodulin-depleted membranes, but the inhibitory effect of lithium could be restored by addition of exogenous calmodulin. The present results suggest that lithium might influence the interaction of calmodulin with the enzyme and/or interfere with the divalent cation site(s) on the adenylate cyclase system. PMID- 3031641 TI - [Regional odontodysplasia]. PMID- 3031642 TI - [Value and prognostic significance of alpha-lactalbumin (ALA), lactoferrin (Lfr) and HMFG-2 in primary breast cancers. An immunohistochemical study]. PMID- 3031643 TI - A specific endpoint assay for ubiquitin. AB - Simple endpoint assays for free ubiquitin (Ub) and for the Ub-activating enzyme are described. The method for measuring Ub makes use of the reaction of iodoacetamide-treated Ub-activating enzyme (E): [3H]ATP + Ub + E----E X [3H]AMP Ub + PPi and PPi----2Pi (in the presence of pyrophosphatase). The Ub is then measured by determining the acid-insoluble radioactivity. The reaction is accompanied by a slow enzyme-catalyzed hydrolysis of the complex to AMP plus Ub. The presence of ubiquitin-activating enzyme in excess of Ub by approximately equal to 0.1 microM assures that the steady state will be close to the endpoint for total Ub. A preparation of the activating enzyme from human erythrocytes that does not depend on affinity chromatography is described. Several applications of the assay are presented. PMID- 3031646 TI - Human salivary gustin is a potent activator of calmodulin-dependent brain phosphodiesterase. AB - Human salivary gustin stimulated activity of brain calmodulin-dependent cyclic nucleotide phosphodiesterase (cAMP PDEase; 3',5'-cyclic-nucleotide phosphodiesterase, EC 3.1.4.17) in a dose-dependent manner in the absence of calmodulin. At physiological levels found in human saliva, gustin activated cAMP PDEase 5- to 6-fold. Activation of PDEase occurred with as little as 500 ng of gustin. Comparative sensitivity of activation of PDEase by gustin was intermediate between calmodulin and lysophosphatidylcholine with maximal activation and half-maximal activation (indicated in parentheses) at 3 X 10(-8) M (4.3 X 10(-9) M), 3.4 X 10(-6) M (3.4 X 10(-7) M), and 2.5 X 10(-3) M (4.0 X 10( 5) M) for calmodulin, gustin, and lysophosphatidylcholine, respectively. No other major salivary protein activated PDEase. Anticalmodulin antibody completely inhibited calmodulin-activated cAMP PDEase activity, but the antibody had no effect on gustin-activated cAMP PDEase activity. A sensitive calmodulin RIA indicated that no calmodulin was detected in any gustin preparation that activated cAMP PDEase. Both gustin and calmodulin rendered cAMP PDEase thermally labile to a similar extent and increased Vmax without affecting the apparent Km for the substrate cAMP. Activation by gustin and calmodulin was unaffected by lubrol-PX, trypsin inhibitor, pepstatin A, or leupeptin. In the presence of 1 mM EGTA, gustin activated cAMP PDE 5- to 6-fold, but the activating ability was completely lost after gustin was heated at 100 degrees C for 5 min. In contrast, calmodulin lost all activating ability in the presence of 1 mM EGTA, whereas heating calmodulin at 100 degrees C for 5 min did not affect its activation of cAMP PDEase. Lysophosphatidylcholine-activation of cAMP PDEase, like gustin activation, was unaffected by EGTA, but lysophosphatidylcholine-activation of cAMP PDEase, like calmodulin activation, was unaffected by heating at 100 degrees C for 5 min. PMID- 3031645 TI - Identification of a peptide binding protein that plays a role in antigen presentation. AB - The helper T-cell response to globular proteins appears, in general, to require intracellular processing of the antigen, such that a peptide fragment containing the T-cell antigenic determinant is released and transported to and held on the surface of an Ia-expressing, antigen-presenting cell. However, the molecular details underlying these phenomena are largely unknown. The means by which antigenic peptides are anchored on the antigen-presenting cell surface was investigated. A cell surface protein is identified that was isolated by its ability to bind to a 24-amino acid peptide fragment of pigeon cytochrome c, residues 81-104, containing the major antigenic determinant for B10.A mouse T cells. This peptide binding protein, purified from [35S]methionine-labeled cells, appears as two discrete bands of approximately equal to 72 and 74 kDa after NaDodSO4/PAGE. The protein can be eluted from the peptide affinity column with equivalent concentrations of either the antigenic pigeon cytochrome c peptide or the corresponding nonantigenic peptide of mouse cytochrome c. However, it does not bind to the native cytochromes c, either of pigeon or mouse, and thus the protein appears to recognize some structure available only in the free peptides. This protein plays a role in antigen presentation as evidenced by the ability of rabbit antibodies raised against it to block the activation of an antigen specific T-cell hybrid by antigen-presenting cells and pigeon cytochrome c. Its expression is not major histocompatibility complex-restricted in that the blocking activity of the antisera can be absorbed on spleen cells from mice of different haplotypes. This peptide binding protein can be isolated from a variety of cell types, including B cells, T cells, and fibroblasts. The anchoring of processed peptides on the cell surface by such a protein may play a role in antigen presentation--facilitating the interaction of antigenic peptides with Ia and/or the T-cell receptor. PMID- 3031644 TI - Coordinate tropic hormone regulation of mRNAs for insulin-like growth factor II and the cholesterol side-chain-cleavage enzyme, P450scc [corrected], in human steroidogenic tissues. AB - Insulin-like growth factors (IGFs) are single-chain polypeptides important for cell proliferation and growth. IGFs are produced in several tissues, suggesting that they function in a paracrine or autocrine fashion as well as functioning as endocrine hormones. We studied the hormonal regulation of IGF-I and IGF-II mRNA in human steroidogenic tissues. In cultured human ovarian granulosa cells, follicle-stimulating hormone, human chorionic gonadotropin, and dibutyryl cAMP increased IGF-II mRNA, but corticotropin [adrenocorticotropic hormone (ACTH)], chorionic somatomammotropin, growth hormone, prolactin, dexamethasone, estradiol, and progesterone had no effect. In cultured human fetal adrenal cells, ACTH and dibutyryl cAMP increased IGF-II mRNA accumulation, but human chorionic gonadotropin and angiotensin II did not. The same five size species of IGF-II mRNA were detected in transfer blots of RNA from granulosa cells and fetal adrenal cells, and all of these increased after hormonal stimuli. Dibutyryl cAMP also increased IGF-II mRNA accumulation in cultured human placental cells. Accumulation of mRNA for the cholesterol side-chain-cleavage monooxygenase [P450scc [corrected]; cholesterol, reduced-adrenal-ferredoxin:oxygen oxidoreductase (side-chain-cleaving), EC 1.14.15.6] was regulated in parallel with IGF-II mRNA in all these steroidogenic tissues. IGF-I mRNA was not detected in transfer blots of these RNAs, and the minimal amounts detected in dot blots showed no detectable change after any of the hormonal stimuli studied. The data indicate that the IGF-II gene is expressed in human steroidogenic tissues and is regulated by cAMP. These data suggest that IGF-II may act in an autocrine or paracrine fashion to stimulate the adrenal and gonadal growth stimulated by ACTH and gonadotropins, respectively. PMID- 3031647 TI - Application of a protein-blotting procedure to the study of human glucocorticoid receptor interactions with DNA. AB - To exert their effects, glucocorticoid receptor complexes interact selectively with DNA sequences known as glucocorticoid regulatory elements. We have studied the interaction between human glucocorticoid receptors and mouse mammary tumor virus (MMTV) DNA by means of a procedure that permits analysis after immobilization of the receptor on nitrocellulose. Proteins from crude cytosolic or nuclear extracts were electrophoresed on NaDodSO4/PAGE gels, soaked in a urea buffer to remove NaDodSO4, transferred to nitrocellulose, and probed with nick translated MMTV [32P]DNA in a 5% nonfat dry milk buffer, which minimizes nonselective DNA-protein interactions. We present evidence that MMTV [32P]DNA interacts selectively with the glucocorticoid receptor. These data include comigration of [3H]dexamethasone mesylate-labeled band and bound MMTV [32P]DNA on gel electrophoresis systems; localization of DNA-binding activity in the cytosol of cells incubated with steroid at 0 degrees C and in the nucleus and cytosol of cells incubated at 37 degrees C; binding of the MMTV DNA to highly purified receptor; and absence of MMTV DNA binding activity in extracts from cells whose receptor has been down-regulated. Furthermore, glucocorticoid receptors analyzed under these conditions exhibit selective binding to DNA fragments that contain glucocorticoid regulatory elements. PMID- 3031648 TI - Uptake of norepinephrine and related catecholamines by cultured chromaffin cells: characterization of cocaine-sensitive and -insensitive plasma membrane transport sites. AB - Norepinephrine and its closely related analogues, dopamine and epinephrine, are transported into chromaffin cells in culture by two distinct types of sites on the plasma membrane: one is sensitive to cocaine while the other is not. The cocaine-sensitive site has a high affinity for catecholamines and depends on sodium in the medium. The apparent Km for norepinephrine uptake by the cocaine sensitive site is 5.8 microM when determined in the presence of 118 mM NaCl, obtained using nonlinear least-square curve fitting. Detailed kinetic analysis has also shown cocaine to be a competitive inhibitor of norepinephrine uptake with an apparent Ki of ca. 1 microM. This site is blocked by a series of tricyclic antidepressant drugs with relative potencies characteristic of norepinephrine transport sites in neurons. In contrast, the cocaine-insensitive site(s) have a low affinity for norepinephrine (apparent Km, approximately 88 microM) and are also able to transport catecholamine analogues such as dimethyl epinephrine and isoproterenol, which have bulky groups attached to the amine moiety. Transport of norepinephrine at both sites is blocked by low temperature, by mitochondrial uncouplers, and by other metabolic inhibitors. Both of these transport sites in the chromaffin cell plasma membrane, therefore, appear to be different from the well-characterized catecholamine transport sites in the chromaffin granule membrane on the basis of substrate specificity and their sensitivity to inhibitors. PMID- 3031649 TI - Evidence for altered DNA conformations in the simian virus 40 genome: site specific DNA cleavage by the chiral complex lambda-tris(4,7-diphenyl-1,10 phenanthroline)cobalt(III). AB - lambda-Tris(4,7-diphenyl-1,10-phenanthroline)cobalt(III), a photoactivated DNA cleaving agent, is a small molecular probe of DNA structure. Because of its chirality, the complex cannot bind to regular right-handed B-form DNA but exhibits site-specific cleavage along the polymer strand at conformationally distinct sites such as those in a left-handed conformation. Both coarse and higher resolution mapping experiments using the chiral cobalt complex indicate intriguing conformational variations along the simian virus 40 genome. Highly specific cleavage is evident in the enhancer and promoter blocks and in the region downstream of 3' termini. A specific cleavage pattern borders an alternating purine/pyrimidine stretch within the enhancer, which was found earlier to bind anti-Z-DNA antibodies. Throughout the simian virus 40 genome, variations in structure delineated with the cobalt complex appear to correlate with regions important for control of gene expression. PMID- 3031650 TI - Multicopy plasmid with a structure related to the polyoma virus genome. AB - In a subclone derived from mouse L(tk-) cells, we found a plasmid present in a high copy number (greater than 5000 copies per cell) that was stably maintained extrachromosomally without any cytopathic effect to the host cells. This plasmid, termed L factor, has two forms: 5.3 and 5.5 kilobase pairs. DNA sequencing and restriction enzyme mapping showed that, although the structure contains DNA sequences common to polyoma virus, plasmid sequences belonging to the regulatory region (the enhancer region) and other regions are quite different from those in polyoma. In cells bearing the plasmid, we detected a low level of material that cross-reacts with antibody to polyoma tumor antigens, suggesting that the plasmids replicate and are maintained in the cells by a mechanism different from that functioning during propagation following infection of papovaviruses. PMID- 3031651 TI - Structural domains in phage Mu transposase: identification of the site-specific DNA-binding domain. AB - Limited proteolysis of phage Mu transposase with three proteases of differing specificities produced a common pattern of fragmentation. The fragments were mapped by using a combination of immunoblotting and amino acid sequence analysis. Our results suggest that the transposase molecule is organized principally into three domains: an amino-terminal domain of molecular mass 30 kDa, a core region of approximately 35 kDa, and a carboxyl-terminal domain of approximately 10 kDa. The amino-terminal domain has at least two additional sites that are partially accessible to proteases. Filter binding and nuclease protection studies were done to determine the functions of the isolated domains. Site-specific binding to Mu DNA was localized to the amino-terminal domain. The core domain showed nonspecific DNA-binding activity. PMID- 3031652 TI - pen repeat sequences are GGN clusters and encode a glycine-rich domain in a Drosophila cDNA homologous to the rat helix destabilizing protein. AB - Several cDNA clones that contain the pen repeat have been isolated and sequenced; pen consists of clusters of GGN triplets, where N can be any nucleotide. Some of the pen repeat sequences are found within long open reading frames in which they encode oligoglycine stretches. For one of the clones, the deduced amino acid sequence of the entire open reading frame, especially in the region preceding the glycine-rich domain, shows strong homology to the rat helix destabilizing protein [Cobianchi, F., SenGupta, D. N., Zmudzka, B. Z. & Wilson, S. H. (1986) J. Biol. Chem. 261, 3536-3543]. The rat protein and homologs in other organisms are single stranded nucleic acid binding proteins, some of which are major components of heterogeneous nuclear ribonucleoprotein particles. We suggest that we have cloned a cDNA encoding a Drosophila single-stranded nucleic acid binding protein. PMID- 3031653 TI - Ubiquitin-aldehyde: a general inhibitor of ubiquitin-recycling processes. AB - The generation and characterization of ubiquitin (Ub)-aldehyde, a potent inhibitor of Ub-C-terminal hydrolase, has previously been reported. We now examine the action of this compound on the Ub-mediated proteolytic pathway using the system derived from rabbit reticulocytes. Addition of Ub-aldehyde was found to strongly inhibit breakdown of added 125I-labeled lysozyme, but inhibition was overcome by increasing concentrations of Ub. The following evidence shows the effect of Ub-aldehyde on protein breakdown to be indirectly caused by its interference with the recycling of Ub, leading to exhaustion of the supply of free Ub: Ub-aldehyde markedly increased the accumulation of Ub-protein conjugates coincident with a much decreased rate of conjugate breakdown. release of Ub from isolated Ub-protein conjugates in the absence of ATP (and therefore not coupled to protein degradation) is markedly inhibited by Ub-aldehyde. On the other hand, the ATP-dependent degradation of the protein moiety of Ub conjugates, which is an integral part of the proteolytic process, is not inhibited by this agent. Direct measurement of levels of free Ub showed a rapid disappearance caused by the inhibitor. The Ub is found to be distributed in derivatives of a wide range of molecular weight classes. It thus seems that Ub-aldehyde, previously demonstrated to inhibit the hydrolysis of Ub conjugates of small molecules, also inhibits the activity of a series of enzymes that regenerate free Ub from adducts with proteins and intermediates in protein breakdown. PMID- 3031654 TI - Replication of simian virus 40 origin-containing DNA in vitro with purified proteins. AB - Simian virus 40 (SV40) DNA replication dependent on the SV40 origin of replication and the SV40 large tumor (T) antigen has been reconstituted in vitro with purified protein components isolated from HeLa cells. In addition to SV40 T antigen, these components included the DNA polymerase alpha-primase complex, topoisomerase I, and a fraction that contained a single-stranded DNA binding protein. The latter protein, which sediments at 5.1 S on glycerol gradients and copurifies with two major protein species of 72 and 76 kDa, was isolated solely by its ability to support SV40 DNA replication. The purified system retained the species-specific DNA polymerase alpha-primase requirement previously observed with crude fractions; the complex from HeLa cells supported SV40 replication, whereas that from calf thymus and mouse cells did not. DNA containing the polyomavirus origin of replication was replicated in a system containing polyomavirus T antigen, the HeLa single-stranded DNA binding protein-containing fraction, and DNA polymerase alpha-primase complex from mouse, but not HeLa, cells. While crude fractions yielded closed circular duplex DNA, none was detected with the purified system. Nevertheless, the addition of a crude fraction to the purified system yielded closed circular monomer products. PMID- 3031655 TI - Multiple forms of basic fibroblast growth factor: amino-terminal cleavages by tumor cell- and brain cell-derived acid proteinases. AB - Basic fibroblast growth factor (FGF) was purified by heparin-Sepharose chromatography from two sources, brain and hepatoma cells. Brain cell-derived basic FGF (brFGF) and hepatoma cell-derived basic FGF (heFGF) were found to exist in multiple forms whose molecular weights depended on whether they were extracted from their respective tissue or cells at neutral or acid pH. When extracted at pH 7.0 brFGF and heFGF comigrated on NaDodSO4/PAGE with a Mr of approximately 18,400. When extracted at pHs 3.5-4.5, acid proteinases cleaved brFGF and heFGF to lower molecular weight forms but to different extents. brFGF was cleaved to a Mr 18,000 form at acid pH by a brain-derived acid proteinase that could be inhibited by pepstatin. heFGF was cleaved mostly to a Mr 16,500 form at acid pH by a hepatoma cell-derived acid proteinase that was inhibited by leupeptin. Electrophoretic transfer blot analysis using site-specific anti-FGF antibodies suggested that the cleavages occurred at the amino-terminal ends of brFGF and heFGF. Cleavage to lower molecular weight forms of brFGF and heFGF did not affect growth factor activity or chromatographic behavior on heparin-Sepharose columns. PMID- 3031656 TI - Cloning and analysis of the promotor region of the human fibronectin gene. AB - Human fibronectin (FN) genomic clones were isolated by screening a human genomic library with a 75-base oligonucleotide. The sequence of the oligonucleotide corresponds to a region near the 5' end of the human FN cDNA clone pFH6 that contains the amino-terminal coding sequences but does not extend to the 5' end of the mRNA [Kornblihtt, A. R., Umezawa, K., Vibe-Pedersen, K. & Baralle, F. E. (1985) EMBO J. 4, 1755-1759]. The 5' end of the FN gene is found on a 3.7 kilobase-pair EcoRI fragment that contains about 2.7 kilobase pairs of flanking sequence. The first exon is 414 base pairs long, with a 5' untranslated region of 267 base pairs. As deduced on the basis of the position of the initiation codon, FN is synthesized with a 31-residue amino acid extension on the amino terminus that is not present in the mature polypeptide. This amino-terminal extension appears to contain both a signal peptide and a propeptide. The first 200 base pairs of 5'-flanking sequence is very G + C rich. Upstream of this the sequence becomes relatively A + T rich. The sequence ATATAA is found at -25 and the sequence CAAT is present at -150. The sequence GGGGCGGGGC at -102 exhibits homology to the binding site for the transcription factor SP1, and the sequence TGACGTCA at -173 exhibits homology to 5'-flanking sequences important for induction by cAMP. PMID- 3031657 TI - Isolation of the tissue factor inhibitor produced by HepG2 hepatoma cells. AB - Progressive inhibition of tissue factor activity occurs upon its addition to human plasma (serum). This process requires the presence of factor VII(a), facto X(a), Ca2+, and another component in plasma that we have called the tissue factor inhibitor (TFI). A TFI secreted by HepG2 cells (human hepatoma cell line) was isolated from serum-free conditioned medium in a four-step procedure including CdCl2 precipitation, diisopropylphosphoryl-factor Xa affinity chromatography, Sephadex G-75 superfine gel filtration, and Mono Q ion-exchange chromatography. The purified TFI contained a predominant band at Mr 38,000 on NaDodSO4/polyacrylamide gel electrophoresis that comigrates with inhibitory activity. Like the activity present in plasma, this TFI requires the presence of factor VII(a), factor X(a), and Ca2+ to express inhibitory activity. Its specific activity (assuming an extinction coefficient of 10 at 280 nM, for a 1-cm path length through a 1% solution) was 9800 units/mg of protein, where 1 unit of TFI activity was defined as that present in 1 ml of normal pooled serum. PMID- 3031660 TI - Interleukin 2 binding molecule distinct from the Tac protein: analysis of its role in formation of high-affinity receptors. AB - Interleukin 2 (IL-2) receptors on activated T cells exist in high- and low affinity configurations, both of which share a ligand-binding component known as the Tac protein. Although almost all binding of IL-2 to such cells was inhibited by an antibody to Tac, the predominant component of binding on the natural killer (NK)-like cell line YT was resistant to this reagent. The ligand-binding component on YT cells also differed from Tac in its affinity constant (Kd approximately 8.2 X 10(-10) M vs. Kd approximately equal to 1.1 X 10(-8) M for low-affinity Tac sites) and in its susceptibility to inhibition by certain antibodies to IL-2. When the YT cells were stimulated in a manner that induced expression of the Tac protein, the IL-2 binding sites were converted to a high affinity configuration (Kd approximately 1.8 X 10(-11) M). Thus, the original binding component on unstimulated YT cells appeared to combine with Tac and IL-2 to produce a high-affinity receptor complex. Use of bifunctional crosslinking agents following ligand binding to unstimulated YT cells yielded covalent IL-2 receptor complexes of 83 and 90 kDa. These complexes were similar in size to those derived from high-affinity receptors on activated T cells and shared a similar fragmentation pattern upon proteolysis. These results demonstrate the existence of a second IL-2 binding component in addition to the Tac protein and suggest that this component combines with Tac and IL-2 to form high-affinity receptor sites. PMID- 3031659 TI - Involvement of chromosome X in primary cytogenetic change in human neoplasia: nonrandom translocation in synovial sarcoma. AB - A translocation that involves chromosome X (band p11.2) and chromosome 18 (band q11.2) was observed in short-term in vitro cultures of cells from five synovial sarcomas and one malignant fibrous histiocytoma. In four of these tumors, the translocation t(X;18)(p11.2;q11.2) was reciprocal. The two other tumors had complex translocations: t(X;18;21)(p11.2;q11.2;p13) and t(X;15;18)(p11.2;q23;q11.2). A translocation between chromosomes X and 18 was not detected in other histological types of soft tissue sarcoma. The X;18 rearrangement appears to characterize the synovial sarcoma and is the first description of a primary, nonrandom change in the sex chromosome of a human solid tumor. PMID- 3031658 TI - Herpes simplex virus-infected cells contain a function(s) that destabilizes both host and viral mRNAs. AB - The herpes simplex virus virion contains a function that mediates the shutoff of host-protein synthesis and the degradation of host mRNA. Viral mutants affected in this function (vhs mutants) have previously been derived. Cells infected with these mutants exhibit a more stable synthesis of host as well as the immediate early (alpha)-viral proteins. We now show that a function associated with purified virions of the wild-type virus reduces the half-life of host and alpha mRNAs, whereas purified vhs-1 mutant virions lack this activity. The functional half-life of many early (beta)- and late (gamma)-viral mRNAs is also prolonged in mutant virus infections. These studies suggest that the wild-type virion brings into cells a function that indiscriminately reduces the half-life of both host and viral transcripts and that the early translational shutoff of the host is a consequence of this function. This function may facilitate rapid transitions in the expression of groups of genes that are transcriptionally turned on at different times after infection. PMID- 3031661 TI - Neuropeptide Y inhibits cholinergic transmission in the isolated guinea pig colon: mediation through alpha-adrenergic receptors. AB - Neuropeptide Y, a member of the pancreatic polypeptide family, caused dose dependent relaxation of guinea pig colon longitudinal muscle. This inhibitory effect was unaffected by hexamethonium but was abolished by atropine and tetrodotoxin, suggesting that neuropeptide Y is acting via postganglionic cholinergic neurons. Studies utilizing alpha-adrenergic and dopamine receptor antagonists revealed that neuropeptide Y-mediated relaxation was blocked only by the alpha 2 antagonist yohimbine. Neuropeptide Y also antagonized muscle response to electric field stimulation that was reversed by yohimbine. Additional studies with muscle slices incubated with [3H]norepinephrine or [3H]choline showed that neuropeptide Y stimulated norepinephrine release from sympathetic nerves, which, in turn, inhibited the release of acetylcholine via alpha 2 receptors located on postganglionic nerves. This pathway provides a mechanism for neuropeptide modulation of classical neurotransmitter function. PMID- 3031662 TI - cDNA cloning of human myeloperoxidase: decrease in myeloperoxidase mRNA upon induction of HL-60 cells. AB - Myeloperoxidase (MPO), the most abundant neutrophil protein, is a bacteriocidal component of the primary granules and a critical marker in distinguishing acute myelogenous leukemia from acute lymphoid leukemia. A cDNA clone for human MPO was isolated by immunologic screening of human hematopoietic lambda gt11 expression vector libraries with specific anti-MPO antibody. The identity of the cDNA clone was confirmed by finding that epitope-selected antibody against this clone recognizes purified MPO and MPO in human promyelocytic (HL-60) cell lysates by immunoblot analysis, and that hybrid selection of HL-60 mRNA with this cDNA clone and translation in vitro results in the synthesis of an 80-kDa protein recognized by the anti-MPO antiserum. RNA blot analysis with this MPO cDNA clone detects hybridization to two polyadenylylated transcripts of approximately 3.6 and approximately 2.9 kilobases in HL-60 cells. No hybridization is detected to human placenta mRNA. Upon induction of HL-60 cells to differentiate by incubation for 4 days with dimethyl sulfoxide, a drastic decrease in the hybridization intensity of these two bands is seen. This is consistent with previous data suggesting a decrease in MPO synthesis upon such induction of these cells. The MPO cDNA should be useful for further molecular and genetic characterization of MPO and its expression and biosynthesis in normal and leukemic granulocytic differentiation. PMID- 3031663 TI - Structure and expression of the rat neuropeptide Y gene. AB - Neuropeptide Y is a 36-amino acid peptide that is abundant throughout the mammalian nervous system. It belongs to the same family of carboxyl-terminally amidated peptides as pancreatic polypeptide and peptide YY. We describe here the gene encoding the rat neuropeptide Y precursor. The gene spans 7.2 kilobase pairs and contains four exons. The exon organization is identical to the pancreatic polypeptide gene, although the amino acid sequences of the neuropeptide Y and pancreatic polypeptide precursors differ extensively. The predicted amino acid sequence of mature rat neuropeptide Y is identical to the human sequence. Also the sequence of the 30-amino acid carboxyl-terminal peptide of preproneuropeptide Y is highly conserved, which suggests that it is functionally important. Two neuropeptide Y alleles were found to differ at nine positions in 2.5 kilobase pairs at the 5' portion of the gene. No exon difference was found. One nucleotide substitution close to the gene promoter may influence the regulation of expression. Neuropeptide Y mRNA was found in all rat brain subregions tested, which shows that neuropeptide Y is synthesized throughout the brain. Developmentally, mRNA was detected in the rat brain as early as embryonic day 16 and increased rapidly to adult levels. The level of neuropeptide Y mRNA was also studied in several rat peripheral organs. Unexpectedly high levels were observed in heart and spleen. This mRNA may be synthesized in intrinsic ganglia and non neuronal cells, respectively. PMID- 3031665 TI - Occurrence in Saccharomyces cerevisiae of a gene homologous to the cDNA coding for the alpha subunit of mammalian G proteins. AB - From cross-hybridization studies with cDNAs that code for the alpha subunits of rat brain guanine nucleotide-binding regulatory (G) proteins, we have isolated a gene from yeast Saccharomyces cerevisiae encoding an amino acid sequence that is highly homologous to the alpha subunit of the G protein that mediates inhibition of adenylate cyclase (Gi alpha) from rat brain. The gene, tentatively designated as GPA1, contains a contiguous, single open reading frame of 1416 nucleotides that codes for a protein of 472 amino acids with a calculated Mr of 54,075. The predicted amino acid sequence of the protein encoded by the GPA1 gene (tentatively designated as G protein 1 alpha or GP1 alpha) is remarkably homologous to the amino acid sequence of rat brain Gi alpha and the alpha subunit of the G protein of unknown function (Go alpha); the primary structure of the sites for GTP hydrolysis as well as GTP interaction are nearly identical. The main difference in the molecular sizes of yeast GP1 alpha (472 amino acids) and rat brain Gi alpha (355 amino acids) is due to the presence of a stretch of 110 extra amino acid residues in yeast GP1 alpha, which are inserted near the NH2 terminal one-third of mammalian Gi alpha. From blot-hybridization analysis, the size of the GP1 alpha mRNA was estimated as 1.7 kilobases. PMID- 3031664 TI - Two promoters, one inducible and one constitutive, control transcription of the Streptomyces lividans galactose operon. AB - Galactose utilization in Streptomyces lividans was shown to be controlled by an operon that is induced in the presence of galactose and repressed by glucose. Two promoters, galP1 and galP2, which direct transcription of two distinct polycistronic transcripts, have been identified. galP1 is located immediately upstream of the operon and is induced in the presence of galactose. This promoter directs transcription of the galT, galE, and galK genes. The second promoter, galP2, is located within the operon just upstream of the galE gene. This promoter is responsible for constitutive transcription of the galE and galK genes. Comparison of the S. lividans gal operon to the Escherichia coli gal operon indicates the presence of a constitutive promoter positioned upstream of galE in both operons. We suggest that coupling the operon's constitutive promoter to the galE gene fulfills a physiological requirement for constitutive UDPgalactose 4 epimerase expression in Streptomyces. PMID- 3031666 TI - Protein glycosylation in yeast: transcript heterogeneity of the ALG7 gene. AB - The first enzyme in the lipid-linked oligosaccharide biosynthetic pathway, UDP-N acetylglucosamine-dolichyl-phosphate N-acetylglucosaminephosphotransferase (UDP-N acetyl-D-glucosamine:dolichyl-phosphate-N-acetyl- D glucosaminephosphotransferase, EC 2.7.8.15), is encoded by the ALG7 gene. We show that this gene is essential for cell growth, since a null mutation constructed with standard gene disruption techniques results in cell lethality. The ALG7 gene is transcribed into two major messages, approximately 1.38 and 1.56 kilobase pairs (kbp) in size, and this heterogeneity has been mapped to the 3' untranslated region. Two sets of tripartite sequences implicated in transcription termination begin 15 bp and 256 bp past the translation stop codon, TGA. The ratios of the two major transcripts change with gene dosage, with the longer mRNA becoming more abundant in cells containing higher levels of the ALG7 gene. Changes in transcript ratios are also observed in mutants defective in lipid linked sugar-donor biosynthesis. In addition, there is 5' heterogeneity in the ALG7 mRNAs. The transcripts start at four initiation sites located within a 20-bp region. Two potentially functional TATA elements have been identified at positions -157 and -139, which may be involved in initiation from multiple sites. These features point to numerous factors that may be involved in the regulation of the expression of the ALG7 gene. PMID- 3031668 TI - Cosmid vectors for rapid genomic walking, restriction mapping, and gene transfer. AB - We have designed cosmid vectors for rapid genomic "walking" and restriction mapping. These vectors contain the transcription promoters from either bacteriophage SP6, T7, or T3 flanking a unique BamHI cloning site. Mammalian expression modules encoding the dominant marker neomycin phosphotransferase or the amplifiable dihydrofolate reductase gene expressed from SV40 promoters were inserted for use in gene transfer studies. Restriction sites for the enzymes Not I and Sfi I, which cut mammalian DNA very infrequently, have been engineered near the transcriptional promoters to enable the excision of most inserts as single, full-length fragments. Genomic libraries representative of mouse, human, and hamster genomes were constructed by inserting 33- to 44-kilobase-pair (kbp) DNA fragments, generated by partial cleavage of genomic DNA with Mbo I or Sau3A, into the unique BamHI site. Digestion of recombinant cosmids with restriction enzymes that cleave frequently but do not disrupt the transcriptional promoters generates two small DNA templates for the synthesis of end-specific RNA probes to facilitate directional "walking." Cosmid restriction maps can be determined rapidly by one of several methods. The cosmids and methods we describe should have wide utility in determining the functional and structural organization of complex eukaryotic genomes and for physically linking distant genetic loci. PMID- 3031667 TI - Expression of human class II major histocompatibility complex antigens using retrovirus vectors. AB - Retrovirus vectors [direct orientation (DO) vectors] that permit the simultaneous expression of an inserted protein-coding sequence and a dominant-acting selectable marker have been constructed. In these vectors, an internal simian virus 40 or human metallothionein promoter sequence serves to drive the expression of the bacterial neomycin phosphotransferase or guanine-xanthine phosphoribosyltransferase genes, whereas the viral long terminal repeat sequences are utilized to promote expression of inserted sequences. In some of the vectors, the viral 5' splice site, normally used in the biogenesis of the subgenomic env encoding mRNA, has been eliminated. These vectors yield high transient and stable titers of virus after transfection of viral packaging cell lines, show little or no depression of virus titer with a variety of inserts, and faithfully transmit recombinant proviral sequences to recipient cells. To characterize the expression potential of these vectors, a variety of inserts encoding the alpha and beta subunits of the human major histocompatibility complex class II antigen HLA-DR have been introduced into these vectors. NIH 3T3 cells infected by viruses containing HLA-DR alpha or beta cDNAs express these proteins as shown by immunoprecipitation of metabolically labeled extracts. In addition, through the sequential infection of cells with retrovirus constructions expressing two different selectable markers, both subunits of the class II antigen have been introduced into NIH 3T3 cells. Such infected cells express HLA-DR molecules at the cell surface. PMID- 3031669 TI - Pathway for inositol 1,3,4-trisphosphate and 1,4-bisphosphate metabolism. AB - We prepared [3H]inositol-,3-[32P]phosphate-and 4-[32P]phosphate-labeled inositol phosphate substrates to investigate the metabolism of inositol 1,3,4 trisphosphate and inositol 1,4-bisphosphate. In crude extracts of calf brain, inositol 1,3,4-trisphosphate is first converted to inositol 3,4-bisphosphate, then the inositol 3,4-bisphosphate intermediate is further converted to inositol 3-phosphate. Similarly, inositol 1,4-bisphosphate is converted to inositol 4 phosphate, and no inositol 1-phosphate is formed. We partially purified an enzyme that we tentatively name inositol polyphosphate 1-phosphatase. This cytosolic enzyme converts inositol 1,3,4-trisphosphate to inositol 3,4-bisphosphate and also converts inositol 1,4-bisphosphate to inositol 4-phosphate. The enzyme does not utilize inositol 1,3,4,5-tetrakisphosphate, inositol 1,4,5-trisphosphate, or inositol 1-phosphate as substrates. Thus we propose a new scheme for inositol phosphate metabolism. According to this pathway inositol 1,4,5-trisphosphate and inositol 1,4-bisphosphate are degraded to inositol 4-phosphate. Inositol 1 phosphate, which is the major inositol monophosphate formed in stimulated brain, is derived either from phospholipase C cleavage of phosphatidylinositol or from the degradation of inositol cyclic phosphates. PMID- 3031671 TI - Detection of three protein binding sites in the serum-regulated promoter of the human gene encoding the 70-kDa heat shock protein. AB - The basal promoter of the human gene encoding 70-kDa heat shock protein (HSP70) controls maximal transcriptional activity and serum-regulated expression. We demonstrate that the three promoter elements defined by in vivo studies--CCAAT, serum-regulated element (SRE), and "TATA"--correspond to protein-binding sites in vitro. The promoter interactions with protein factors in HeLa cell crude nuclear extracts were detected by an exonuclease III digestion assay. The sequence specificity was demonstrated with promoter probes containing wild-type sequences or unique linker-scanner mutations that alter each of the elements. We suggest that the protein factor binding to the SRE is involved in the serum-regulated expression of the human gene for HSP70. PMID- 3031670 TI - In vivo aminoacylation of human and Xenopus suppressor tRNAs constructed by site specific mutagenesis. AB - Amber suppressor tRNA genes were constructed by site-specific mutagenesis of the anticodons of human lysine-inserting tRNA (tRNA(Lys)) and glutamine-inserting tRNA (tRNA(Gln)) genes, and a Xenopus laevis tyrosine-inserting tRNA (tRNA(Tyr)) gene. As previous in vitro studies in prokaryotes have shown that substitution of nucleotides in the anticodon region can profoundly affect tRNA aminoacylation, it is important to determine whether the mutation affects aminoacylation of these eukaryotic tRNAs. We present a method for quantitating the tRNA aminoacylation in vivo in mammalian cells, and we have determined that the suppressor tRNA(Tyr) is fully aminoacylated and suppressor tRNA(Lys) and tRNA(Gln) are aminoacylated 40 50% and 80%, respectively. This in vivo method of estimating aminoacylation may be applied to other mutations in the tRNA genes. PMID- 3031673 TI - Insulin inhibition of hepatic cAMP-dependent protein kinase: decreased affinity of protein kinase for cAMP and possible differential regulation of intrachain sites 1 and 2. AB - In hepatocytes stimulated with 8-bromo-cAMP, insulin decreases the affinity of the cAMP-dependent protein kinase for cAMP, shifting the Ka without affecting the Vmax activity. This occurs under conditions where cyclic adenine nucleotide concentrations are unchanged. We report here that glycogenolysis stimulated by 8 (4-chlorophenylthio)-cAMP, an analog with 100 times tighter affinity than cAMP for the protein kinase regulatory subunit, was only slightly antagonized by insulin. The tight binding of this analog appears to overcome the protein kinase affinity change induced by insulin. The relative importance of the two intrachain cAMP binding sites of the cAMP-dependent protein kinase regulatory subunit was investigated by using analogs with relative selectivity for each site. Analogs exhibiting preferential binding to site 2 were far less sensitive to insulin antagonism than were analogs binding preferentially at site 1 and less well at site 2. No other property of these analogs, including the rate of hydrolysis by phosphodiesterase, the IC50 for phosphodiesterase, the Ka for protein kinase, or the type I versus type II kinase specificity, could account for the ability of insulin to antagonize glycogenolysis stimulated by these analogs. These data indicate that insulin may act to decrease the affinity of protein kinases for cAMP through a possible regulation of intrachain site 2 binding. PMID- 3031672 TI - Platelet-derived growth factor stimulates phagocytosis and blocks agonist-induced activation of the neutrophil oxidative burst: a possible cellular mechanism to protect against oxygen radical damage. AB - The effect of platelet-derived growth factor (PDGF) on agonist-induced activation of the superoxide-generating oxidative burst in human neutrophils was tested. PDGF had no effect on the resting level of superoxide generation but inhibited both the rate and the extent of fMet-Leu-Phe-stimulated superoxide production in a dose-dependent manner. The concentration required to inhibit the response by 50% was 95 +/- 26 pM (n = 10). PDGF also blocked activation by other receptor mediated agonists such as the complement protein C5a and opsonized zymosan, but not by phorbol myristate acetate or arachidonate, both of which may act at postreceptor sites. The growth factor, however, had no effect on the binding of fMet-Leu-Phe to its receptor. PDGF in concentrations that blocked the oxidative burst stimulated phagocytosis of opsonized latex particles. Thus, PDGF functions as a heterologous "down-regulator" of receptor-mediated activation of the neutrophil oxidative burst and an activator of phagocytosis. A model for a feedback regulatory loop between platelets and neutrophils is proposed. PMID- 3031674 TI - Diphtheria toxin prevents catecholamine desensitization of A431 human epidermoid carcinoma cells. AB - We proposed that a rapidly turning over protein, induced in response to catecholamine stimulation of C6-2B rat astrocytoma cells, inhibits subsequent hormonal activation of adenylate cyclase. Studies upon which our hypothesis is based and confirmatory work in a variety of other cell lines and in vivo have utilized actinomycin D and cycloheximide to inhibit RNA and protein synthesis, respectively. These inhibitors, however, are not specific and have been reported also to interfere with other cellular processes. Diphtheria toxin is a specific protein synthesis inhibitor that acts only by ADP-ribosylating elongation factor 2, thus preventing peptide chain elongation. We thus tested whether diphtheria toxin could prevent catecholamine-induced desensitization in A431 human epidermoid carcinoma cells. The toxin inhibited protein synthesis and altered the time course of isoproterenol-stimulated cAMP accumulation as did the less specific protein synthesis inhibitor cycloheximide. Cellular cAMP content after a 30-min exposure to isoproterenol was similar in control and in toxin-treated cells. However, after 4 hr of treatment with isoproterenol, toxin-treated cells accumulated up to six times more cAMP than controls. When cells or cell-free adenylate cyclase preparations were rechallenged with agonists, toxin-mediated inhibition of protein synthesis prevented desensitization. These results show that diphtheria toxin, a specific inhibitor of protein synthesis, can interfere with the normal physiological regulation of cAMP metabolism in eukaryotic cells and provide compelling evidence that catecholamine stimulation of adenylate cyclase promotes the synthesis of a protein(s) that, in some way, inhibits hormone-stimulated adenylate cyclase. PMID- 3031675 TI - Porphyrins are endogenous ligands for the mitochondrial (peripheral-type) benzodiazepine receptor. AB - "Peripheral-type" benzodiazepine receptors are localized to the outer mitochondrial membrane. We have identified potent competitive inhibitors of these receptors and purified them from human blood and from several rat organs. TLC analysis of the purified inhibitor from erythrocytes displays a single peak of inhibitory activity with an absorbance spectrum identical to hemin. All of the inhibitory activity in extracts of several tissues can be accounted for by their porphyrin and metalloporphyrin content. Pure hemin and protoporphyrin IX competitively inhibit mitochondrial benzodiazepine binding with Ki values of 41 and 15 nM, respectively, and are less active by a factor of 1000 at central-type benzodiazepine receptors. Thus, porphyrins appear to be endogenous ligands for mitochondrial benzodiazepine receptors. PMID- 3031676 TI - Protein kinase C activation enhances cAMP accumulation in Swiss 3T3 cells: inhibition by pertussis toxin. AB - Addition of phorbol 12,13-dibutyrate (PBt2) in the presence of forskolin or cholera toxin caused marked (6- to 8-fold) and rapid accumulation of cAMP in Swiss 3T3 cells. The effect of PBt2 is mediated by protein kinase C because the synthetic diacylglycerol 1-oleoyl-2 acetylglycerol substitutes for PBt2 in enhancing cAMP accumulation and because the enhancing effect of either PBt2 or 1 oleoyl-2-acetylglycerol was prevented by down-regulation of protein kinase C. Vasopressin, which activates protein kinase C but does not directly affect adenylate cyclase in 3T3 cells, also enhanced cAMP accumulation in cells treated with cholera toxin or forskolin. This effect was abolished by down-regulation of protein kinase C. Treatment with pertussis toxin blocked the enhancing effect of PBt2 in a concentration- and time-dependent manner. Pertussis toxin neither prevented protein kinase C activation by PBt2 nor other biologic responses elicited by PBt2. The results presented here suggest an unusual function for a pertussis toxin substrate--namely, coupling protein kinase C activation to cAMP production. PMID- 3031678 TI - Ultrastructural localization of a platelet-derived growth factor/v-sis-related protein(s) in cytoplasm and nucleus of simian sarcoma virus-transformed cells. AB - The subcellular distribution of v-sis-related protein(s) was analyzed in three simian sarcoma virus (SSV)-transformed cell lines with immunofluorescence and protein A-gold labeling techniques using rabbit polyclonal anti-platelet-derived growth factor (PDGF) antisera. Antigenically reactive proteins were recognized in subcellular organelles related to protein synthesis and processing, including polyribosomes, endoplasmic reticulum, and the Golgi apparatus, as well as on the cytoplasmic surface of plasma membranes. Prominent immunoreactive proteins were also shown in association with nuclear chromatin in intact cells and in isolated nuclei using Lowicryl K4M resin embedding techniques. Protein A-gold labeling was markedly reduced in sections of non-SSV-transformed fibroblasts incubated with anti-PDGF and absent from SSV-transformed cells if Epon resin was substituted for Lowicryl in the embedding process or if sections were with irrelevant antisera. Nuclear localization of v-sis-related antigens was confirmed in a nitrocellulose based immunoassay using nuclei isolated from SSV-transformed fibroblasts. Thus, polypeptides recognized antigenically as related to the v-sis gene product not only may be found in subcellular organelles associated with protein synthesis and packaging but also may be found in the nucleus of SSV-transformed cells. These results raise the possibility that v-sis- or PDGF-like proteins may function within the nucleus of SSV-transformed cells. PMID- 3031677 TI - Association of p60src with Triton X-100-resistant cellular structure correlates with morphological transformation. AB - More than 70% of wild-type Rous sarcoma virus p60v-src was found to be associated with a cellular structure resistant to nonionic detergent extraction that consists primarily of cytoskeletal proteins. On the other hand, nontransforming src proteins, including cellular p60c-src, nonmyristoylated forms, and those inactive in protein kinase, were found in the fraction solubilized by the detergent extraction. p60c-src was detergent-soluble even in transformed cells, suggesting that the association of p60v-src is not a result of cell transformation. Analyses with a variety of Rous sarcoma virus mutants showed a good correlation between the degree of association with the detergent-resistant structure and the extent of cell transformation caused by mutant src proteins, suggesting that this association may be significant for the process of cell transformation by Rous sarcoma virus. PMID- 3031679 TI - Loss of heterozygosity in human ductal breast tumors indicates a recessive mutation on chromosome 13. AB - The genotypes at chromosomal loci defined by recombinant DNA probes revealing restriction fragment length polymorphisms were determined in constitutional and tumor tissue from 10 cases of ductal breast cancer: eight premenopausal females and two males. Somatic loss of constitutional heterozygosity was observed at loci on chromosome 13 in primary tumor tissue from three females and one male. In two cases, specific loss of heterozygosity at three distinct genetic loci along the length of the chromosome was observed. In another case, concurrent loss of alleles at loci on chromosomes 2, 13, 14, and 20 was detected, whereas a fourth case showed loss of heterozygosity for chromosomes 5 and 13. In each instance, the data were consistent with loss of one of the homologous chromosomes by mitotic nondisjunction. Analysis of loci on several other chromosomes showed retention of constitutional heterozygosity suggesting the relative specificity of the events. In contrast, similar analyses of other breast cancers, including comedocarcinoma, medullary carcinoma, and juvenile secretory carcinoma, showed no loss of alleles at loci on chromosome 13. These data indicate that the pathogenesis of ductal breast cancer may, in a substantial proportion of cases, involve unmasking of a recessive locus on chromosome 13 and suggest the involvement of such a locus in heritable forms of this disease. PMID- 3031680 TI - Isolation of a cDNA clone and localization of human glutathione S-transferase 2 genes to chromosome band 6p12. AB - The glutathione S-transferases (GST) (glutathione transferase; EC 2.5.1.18) are a family of enzymes responsible for the metabolism of a broad range of xenobiotics and carcinogens. A cDNA clone containing the entire amino acid coding sequence of a human GST-2 subunit has been isolated using a lambda gt11 expression library. The complete nucleotide sequence and a partial restriction map are presented. The subunit is composed of 221 amino acids with a molecular weight of 25,425 before posttranslational modification. The deduced amino acid sequence is rich in lysine, which is consistent with the relatively high pI of GST-2. The human sequence shows considerable homology with the rat Ya and Yc GST sequences but little homology with the rat GSTp and Yb subunit sequences. Southern blots of restriction digests of human DNA indicate that there may be multiple GST-2 genes. In situ hybridization of the cloned cDNA to human chromosomes produces intense labeling only over band p12 on the short arm of chromosome 6 near the centromere. This indicates that the GST-2 gene(s) are located only at this site. PMID- 3031682 TI - Transmission of mitochondrial and chloroplast genomes in crosses of Chlamydomonas. AB - Physical differences between organelle genomes of the interfertile species Chlamydomonas reinhardtii and Chlamydomonas smithii have been used to demonstrate that sexual zygotes transmit chloroplast and mitochondrial DNA from opposite mating types. Processes responsible can be separated functionally and genetically, although both are controlled by mating type. In vegetative diploids, chloroplast and mitochondrial genomes are transmitted biparentally, but a 1 kilobase insert present in the C. smithii mitochondrial genome spreads unidirectionally to all C. reinhardtii genomes in a manner reminiscent of the intron found in the mitochondrial 21S rRNA gene of omega + strains of yeast. PMID- 3031681 TI - S1 nuclease analysis of alpha-globin gene expression in preleukemic patients with acquired hemoglobin H disease after transfer to mouse erythroleukemia cells. AB - The loss of alpha-globin gene transcriptional activity rarely occurs as an acquired abnormality during the evolution of myeloproliferative disease or preleukemia. To test whether the mutation responsible for the loss of alpha globin gene expression (hemoglobin H disease) in these patients is linked with the alpha-globin genes on chromosome 16, we transferred chromosome 16 from preleukemic patients with acquired hemoglobin H disease to mouse erythroleukemia cells and measured the transcriptional activity of the human alpha-globin genes. After transfer to mouse erythroleukemia cells, the expression of human alpha globin genes from the peripheral blood or marrow cells of preleukemic patients with acquired hemoglobin H disease was similar to that of human alpha-globin genes transferred to mouse erythroleukemia cells from normal donors. These data showed that factor(s) in the mouse erythroleukemia cell can genetically complement the alpha-globin gene defect in these preleukemia patients with acquired hemoglobin H disease and suggest that altered expression of a gene in trans to the alpha-globin gene may be responsible for the acquisition of hemoglobin H disease in these patients. PMID- 3031683 TI - Handicapped retroviral vectors efficiently transduce foreign genes into hematopoietic stem cells. AB - Retroviral vectors, designated handicapped, are described. These are genetically defective viruses that allow transfer of nonselectable genes under the transcriptional control of internal promoters. The basic handicapped vector (pHHAM) is derived from Harvey, Abelson, and Moloney murine retroviruses. It contains a 327-base-pair deletion in the 3' long terminal repeat that spans enhancer and promoter sequences. The deletion is successfully transferred to the 5' long terminal repeat after reverse transcription of viral RNA, yielding a provirus incapable of synthesizing viral transcripts. HHAM viruses containing the mouse c-myc gene under the control of immunoglobulin kappa chain gene regulatory elements, along with a selectable gene (neo) driven by a weak promoter (tk), were stably transmitted to cultured mouse B cells. The donor c-myc gene was transcribed from the kappa promoter in these cells. Helper-free virus-producing cell lines were generated at titers favorable for the efficient introduction of HHAM viruses, even without selection, into hematopoietic stem cells from mouse bone marrow. When returned to unirradiated congenic recipient mice, the cells were capable of long-term reconstitution of the myeloid and lymphoid lineages of W/Wv mutants and the lymphoid system of scid mutants. PMID- 3031684 TI - Cell-surface antigens of melanoma recognized by human monoclonal antibodies. AB - Human monoclonal antibodies (mAbs) were derived from lymph node lymphocytes and peripheral blood lymphocytes (PBL) from patients with melanoma. Four methods for generating human mAbs were compared: fusion with human [LICR-LON-HMy-2 (LICR-2)] or mouse (NS-1) cells; transformation by Epstein-Barr virus (EBV); and EBV transformation followed by NS-1 fusion. NS-1 fusion with lymph node lymphocytes resulted in a higher number of growing hybrids than LICR-2 fusion. Virtually no hybrids were obtained from NS-1 or LICR-2 fusions with PBL. EBV transformed lymphocytes from lymph node and peripheral blood with equal efficiency, and the yield of proliferating cultures for antibody screening was more than 10- to 30 fold greater than that obtained by fusion techniques. However, once antibody producing cultures had been identified, stability and clonability of EBV transformed cells were poorer than that of NS-1 hybrid cells. To combine the strengths of both methods, cultures of EBV-transformed cells were fused with NS 1; and hybrid clones were isolated that showed vigorous growth, clonability, and stable antibody secretion. Detailed specificity analysis of the mAbs produced by six of these clones indicated detection of a class 1 (unique) melanoma antigen, a class 3 melanoma antigen, and four ganglioside antigens (GD3, GM3, and two other, as yet uncharacterized, heterophile antigens). PMID- 3031685 TI - Human complement C3b/C4b receptor (CR1) mRNA polymorphism that correlates with the CR1 allelic molecular weight polymorphism. AB - The human C3b/C4b receptor (CR1) is a Mr approximately equal to 200,000 single chain integral membrane glycoprotein of human erythrocytes and leukocytes. It functions both as a receptor for C3b- and C4b-coated ligands and as a regulator of complement activation. Prior structural studies have defined an unusual molecular weight allelic polymorphism in which the allelic products differ in molecular weight by as much as 90,000. On peripheral blood cells there is codominant expression of CR1 gene products of Mr 190,000 (A), 220,000 (B), 160,000 (C), and 250,000 (D). Results of prior biosynthetic and tryptic peptide mapping experiments have suggested that the most likely basis for the allelic molecular weight differences is at the polypeptide level. In order to define further the molecular basis for these molecular weight differences, human CR1 was purified to homogeneity, tryptic peptide fragments were isolated by HPLC and sequenced, oligonucleotide probes were prepared, and a CR1 cDNA was identified. A subclone of this CR1 cDNA was used as a probe of RNA blots of Epstein-Barr virus transformed cell lines expressing the allelic variants. Each allelic variant encodes two distinct transcripts. A mRNA size polymorphism was identified that correlated with the gene product molecular weight polymorphism. This finding, in addition to a prior report of several homologous repeats in CR1, is consistent with the hypothesis that the molecular weight polymorphism is determined at the genomic level and may have been generated by unequal crossing-over. PMID- 3031686 TI - Complete nucleotide sequence of an attenuated hepatitis A virus: comparison with wild-type virus. AB - The complete nucleotide sequence of an attenuated hepatitis A virus, HAV HM-175/7 MK-5, was determined from cloned cDNA. This virus was derived from wild-type HAV HM-175 after 32 passages in African green monkey kidney cells. The resultant cell culture-adapted virus is attenuated for chimpanzees. This virus was passaged an additional three times in monkey kidney cells to obtain sufficient virus for molecular cloning and was designated HM-175/7 MK-5. Three overlapping cDNA clones were obtained that together spanned the entire genome. Comparison of the nucleotide sequence of cDNA from wild-type virus (propagated in marmoset liver in vivo) with attenuated virus (grown in cell culture) showed 24 nucleotide changes distributed throughout the genome. Five base deletions occurred in the 5' noncoding region, and 12 of the 16 base substitutions in the coding region resulted in amino acid changes. Amino acid changes occurred in viral capsid proteins VP1 and VP2 and several of the nonstructural proteins. Thus, a small number of nucleotide changes are responsible for adaptation to cell culture and attenuation of HAV strain HM-175. PMID- 3031687 TI - Molecular structure of mammalian neuropeptide Y: analysis by molecular cloning and computer-aided comparison with crystal structure of avian homologue. AB - Identification and characterization of the cDNA encoding rat neuropeptide Y revealed the nucleotide sequence coding for a 98-amino acid precursor. The deduced amino acid sequence for rat neuropeptide Y is identical to the human peptide and is highly homologous to avian pancreatic polypeptide. The tertiary structure of avian pancreatic polypeptide has been previously derived from crystallographic data by Blundell and coworkers. The homology between neuropeptide Y and avian pancreatic polypeptide preserves all of the residues essential for the maintenance of the tertiary structure. Thus, it has been possible to compute a three-dimensional model of the mammalian neuropeptide, neuropeptide Y, based on the known structure of the avian homologue. This model suggest that neuropeptide preserves a compact tertiary structure characterized by extensive hydrophobic interactions between an N-terminal polyproline-II-like helix and a C-terminal alpha-helix. The model has been used to identify amino acids residing in key positions within this structure and, thereby, to direct future analysis of neuropeptide Y structure-function relationships. PMID- 3031689 TI - Effect of vanadate on amino acid transport in rat jejunum. AB - Vanadate has been reported to inhibit (Na+ + K+)-ATPase of many cells and in some systems to stimulate adenylate cyclase. Since intestinal transport is influenced by these enzymes, we studied the effects of varying concentrations of orthovanadate (VO-4) on alanine transport in the in vitro rat jejunum. At the higher concentrations tested (10(-3) and 10(-2) M) vanadate had a ouabainlike action on alanine transport. It decreased the mucosal-to-serosal flux and the influx of alanine into the intestinal epithelium and it caused a reduction of (Na+ + K+)-ATPase activity of basolateral membranes. The relatively lower vanadate concentration of 10(-4) M increased the influx and the efflux of alanine across the mucosal border of the jejunum. The increase was associated with elevation of cyclic AMP in the intestinal mucosa. The studies suggest the presence of a dual action of vanadate on amino acid transport, a stimulatory effect at low concentration, due to increased adenylate cyclase activity, and an inhibitory effect at higher concentrations, due to a decreased activity of (Na+ + K+)-ATPase. PMID- 3031688 TI - Neurite outgrowth on muscle cell surfaces involves extracellular matrix receptors as well as Ca2+-dependent and -independent cell adhesion molecules. AB - To identify the molecules on the neuronal surface that mediate axonal growth on myotubes, we have examined neurite formation by ciliary neurons grown on myotubes in the absence or presence of specific antibodies. Dramatic inhibition of neurite outgrowth was seen only when antibodies blocked simultaneously the functions of two cell adhesion molecules--neural cell adhesion molecule (N-CAM) and neural Ca2+-dependent CAM (N-Cal-CAM)--and the neuronal receptors for several extracellular matrix (ECM) proteins. Although the antibody used to block ECM receptors (JG22) has been shown to eliminate almost all neurite growth on ECMs, it had only small effects on neurite growth on myotubes, reducing somewhat the length of neurites. Similarly, antibodies to the two CAMs, when used alone, had no detectable effects on neurite length and, when used together, had only small inhibitory effects on neurite growth. Combination of anti-ECM receptor (JG22) with antibodies to either CAM, however, greatly shortened the length of neurites. These results imply that ECM receptors and the CAMs N-CAM and N-Cal-CAM are major macromolecules used by neuronal growth cones for interactions with myotubes. Each provides a distinct mechanism for regulating growth cone motility. PMID- 3031690 TI - Ontogenic corticosteroidogenesis of the domestic fowl: response of isolated adrenocortical cells. AB - Ontogenic adrenocortical function of the domestic was investigated using adrenocortical cells isolated from embryonic chicks (18, 19, 20, and 21 days old) and male and female posthatch birds (1 day, 1 week, and 3 weeks old). Production of the predominant corticosteroids secreted by the chicken adrenal gland, corticosterone, cortisol, and aldosterone, was measured by radioimmunoassay after 2-hr incubation of cells with or without steroidogenic agents. Approaching hatch, basal and maximal ACTH-(1-24) (ACTH)-induced corticosteroid production increased steadily and peaked around 1 day posthatch (5-18 times and 3-9 times, respectively, the production values at 18 days embryonic life). Thereafter, corticosteroid production values decreased steadily to 3 weeks posthatch. Corticosterone predominated over the ages studied: Maximal ACTH-induced corticosterone production averaged 52 and 115 times the production values of aldosterone and cortisol, respectively. In addition, maximal ACTH-induced aldosterone production was roughly 2.2 times greater than cortisol production over the ages studied except for a short-lived, disproportionately greater aldosterone production at 1 day posthatch. In addition to perihatch and age related differences in cellular corticosteroid production, there were also differences in cellular sensitivity to steroidogenic agents as indicated by the differences in half-maximal steroidogenic concentration values (ED50 values) of the steroidogenic agents. Sensitivity to ACTH increased 2.7 times from Day 18 of embryonic life to 1 day posthatch and then decreased steadily to 3 weeks posthatch. In addition, sensitivity to 8-bromo-cAMP (8-Br-cAMP) increased abruptly at 1 day posthatch (nearly 3 times) but then remained constant thereafter. However, a consistent change in cellular sensitivity to 25 hydroxycholesterol was not observed until 3 weeks posthatch (an increase in sensitivity of 3 times that at Day 18 of embryonic life). These data of cellular sensitivity suggest that there were distinct development and maturational alterations in the cellular loci at which ACTH, 8-Br-cAMP, and 25 hydroxycholesterol acted. Thus, during the transition from embryonic to postembryonic life of the domestic fowl, there are alterations in adrenocortical cell steroidogenic capacity and in the function of some cellular loci comprising the corticosteroidogenic pathway. PMID- 3031692 TI - Heterogeneity of rabbit aortic endothelial cells in primary culture. AB - Factor VIII-related antigen (F8-RAg) and angiotensin-converting enzyme (ACE) are accepted diagnostic markers of endothelial cells in culture. However, when we isolated cells from rabbit thoracic aorta (after collagenase treatment and gentle scraping of the intima) and examined them with immunoperoxidase techniques, we observed two cell types which stained specifically for either F8-RAg or ACE, but not both. Each cell type was morphologically distinguishable in primary culture. F8-RAg-positive cells were recognizable in distinct patches as more elongated, tightly apposed, and firmly adherent cells; they exhibited only faint or no staining for ACE and no accumulation of a fluorescent, acetylated low-density lipoprotein probe (DiI-Ac-LDL), another endothelial cell marker. In contrast, ACE positive cells were more rounded, less closely apposed, and grew as strict monolayers that exhibited a characteristic cobblestone appearance at confluence; ACE-positive cells were F8-RAg negative, but demonstrated intense labeling with DiI-Ac-LDL. Subcultures of ACE-positive cells were also stained by anti-rabbit thrombomodulin. PMID- 3031691 TI - Effect of 3,3'-iminodiproprionitrile (IDPN) on corticosteroidogenesis of isolated adrenocortical cells. AB - The neurotoxic agent, 3,3'-iminodiproprionitrile (IDPN), is a disrupter of neurofilament- and intermediate filament-organelle association. In the present study, the effect of IDPN on corticosteroidogenesis was investigated using isolated rat (having few intermediate filaments) and domestic fowl (having abundant intermediate filaments) adrenocortical cells. Cells were incubated with or without steroidogenic agents and precursors and with or without various concentrations of IDPN for 2 hr. IDPN had similar inhibitory potencies (as indicated by the half-maximal inhibitor concentrations (ID50 values] with both rat and domestic fowl cells despite their grossly different intermediate filament content. However, the average ID50 values of IDPN varied with the different steroidogenic agents and precursors used. The average IDPN ID50 values for maximal ACTH- and 8-bromo-cyclic AMP (8-Br-cAMP)-induced corticosterone production were equivalent (49.7 and 45.7 mM, respectively). However, the IDPN ID50 values for maximal ACTH-induced cAMP production, maximal 25 hydroxycholesterol- and pregnenolone-supported corticosterone production, and maximal ACTH- and 8-Br-cAMP-induced protein synthesis varied from 3.7 to 5.4 times the average ID50 values for maximal ACTH- and 8-Br-cAMP-induced corticosterone production. Thus, the inhibitory action of IDPN was not closely linked to the inhibition of ACTH-transmembrane signaling via cAMP, protein synthesis, and steroidogenic enzyme activity. The data suggest that IDPN inhibited corticosteroidogenesis at at a step after cAMP but before cholesterol side-chain cleavage and that the inhibition was not dependent on the presence of intermediate filaments. PMID- 3031693 TI - Component deficiencies. 1. The first component: C1q, C1r, C1s. PMID- 3031694 TI - Blood leukotriene levels during the acute asthma attack in children. AB - Leukotrienes (LT) have been proposed to be important mediators in the etiology of the acute asthma attack (AAA). We therefore studied blood LT levels in 18 children having AAA. Heparinized blood samples were obtained before and after treatment with epinephrine injections and/or metaproterenol inhalations in the emergency room. The samples were acidified and subjected to Sep-pak chromatography. Reverse phase high performance liquid chromatography (RP-HPLC), ultraviolet (UV) spectroscopy and bioassay on guinea pig ileum were used to identify the LT based on comparison to data produced by standard synthetic LT samples. Radioimmunoassay (RIA) was used to further confirm the presence of LT. LT C, D and E were detected in the plasma of children having AAA. Only LT C levels were significantly elevated over control values. The mean blood LT C level of control patients was 1.6 +/- 1.2 nanograms per milliliter (ng/ml, mean +/- SEM) while that of the asthma patients was 73.8 +/- 18.2 ng/ml prior to treatment. After emergency room treatment the asthma patients had a mean blood LT C level of 22.5 +/- 11.7 ng/ml. Lowered levels of LT C accompanied improved clinical condition of the patients. This finding indicates that the AAA in children is associated with elevated blood levels of LT C. PMID- 3031695 TI - Determination of cross tolerance in rat spinal cord using intrathecal infusion via sequential mini-osmotic pumps. AB - Continuous intrathecal (IT) infusion via ALZET mini-osmotic pumps was used to induced spinal tolerance to morphine in the rat. Naloxone (1 mg/kg IP), injected on day 3 of continuous IT morphine (10 micrograms/hr), produced mild withdrawal symptoms in all morphine-treated animals. In rats pretreated with continuous IT morphine (10 micrograms/hr) or saline, systemic morphine (2, 4, 8, 10 and 15 mg/kg IP) produced equivalent, dose-dependent antinociception using the tail flick and paw pressure tests. The rostral and caudal distribution of methylene blue dye in rat spinal cord was determined on days 1-7 of continuous IT infusion. The dye remained localized near the catheter tip throughout infusion; maximum distribution was 1.5 cm rostrally and 1.0 cm caudally. The data indicate that morphine, infused at the rate of 10 micrograms/hr, does not undergo extensive redistribution in the spinal cord. A sequential, double mini-osmotic pump technique for cross tolerance studies in rat spinal cord is described. In rats pretreated with continuous IT norepinephrine for 4 days, the antinociceptive actions of continuous IT morphine were reduced but not significantly different from saline-pretreated animals. These data suggest that morphine, injected into the spinal cord, does not produce behavioural analgesia by activation of local adrenergic systems. PMID- 3031696 TI - Behavioral effects of THC as a function of environment and prior drug experience. AB - Holtzman albino rats were divided into 4 groups, and on 5 consecutive days each group was exposed to one of 4 conditions. The drug-adapted group was given delta 9-tetrahydrocannabinol (THC) (0.0, 0.5, 2.5 or 5.0 mg/kg PO) in their home cages, while the environment-adapted group was given vehicle and placed for one hr in the chamber where they were later tested. The naive group was given vehicle in their home cages and the drug + environment adapted group was given THC and placed in the test chamber. One week later, all rats were given either 0.0, 0.5, 2.5, or 5.0 mg/kg THC and placed in the test chamber where standing, sitting, and behavioral activity were measured. The results showed that the behavioral effects of THC are a function of environmental familiarity in rats who are drug naive but not in rats given prior exposure to THC. PMID- 3031697 TI - Repeated treatment with antidepressant drugs does not affect the benzodiazepine receptors in preincubated membrane preparations from mouse and rat brain. AB - Repeated injections or oral administration of antidepressants: imipramine, amitriptyline and desmethylimipramine (10 mg/kg twice daily for 21 days) did not alter significantly [3H]flunitrazepam binding to frozen-thawed preincubated membranes from the brains of mice or rats. PMID- 3031698 TI - Differential release and effects of LTB4 in guinea-pig lung "in vitro". AB - LTB4 is a potent chemotactic and chemokinetic eicosanoid released by leukocytes during inflammatory reaction; in addition to this it possesses a bronchopulmonary activity in different animal species. Since cysteinyl containing leukotrienes and other eicosanoids have been shown to induce hyperreactivity of pulmonary smooth muscles, we investigated the release of LTB4 from different anatomical structures of guinea-pig lung in vitro and the possible interaction with histamine in these tissues. In fact, hyperreactivity is evidentiated by a synergism between different mediators. The ovalbumin sensitized guinea-pig lung has been brushed in order to separate lung parenchyma from bronchi and vessels which were divided into large and small preparations. The antigen challenge resulted in a statistically significant increase in LTB4 production in large bronchi and vessels, whereas in the other preparations considered the basal levels of the eicosanoid were not modified during the anaphylactic reaction. In parallel with the differential site of LTB4 release, a positive interaction between LTB4 and histamine was only observed in the pulmonary artery. These data suggest that the possible role of LTB4 in different pulmonary diseases is not confined to the airway smooth muscle but it might be related to its capacity to induce vascular hyperreactivity. PMID- 3031699 TI - Lisuride-induced mounting and its modification by drugs active on adrenergic and dopaminergic receptors. AB - Imidazole (IMID), (9.37-75 mg/Kg) and yohimbine (YOH), (0.5-2.5 mg/Kg), strongly potentiated lisuride-induced mounting, scored as a percentage of animals affected and mean number of mounts per animal, while clonidine (150 micrograms/Kg) significantly antagonized the phenomenon. A high (2 mg/Kg) but not a low (50 micrograms/Kg) dose of B-HT 920 and DPI (100 and 500 micrograms/Kg) also inhibited lisuride-induce mounting. While, at present, IMID specific activity on monaminergic system is not yet conclusive, it is demonstrated that, at the doses used, YOH and clonidine are selective alpha 2 antagonist and agonist, respectively; B-HT 920 preferentially stimulates D2 receptors at 50 micrograms/Kg and alpha 2 receptors at 2 mg/Kg; finally DPI, proposed as DAI agonist, also activates alpha 2 receptors. Therefore, in view of the dose-related receptorial selectivity of action of the drugs tested, neurochemical mechanisms on specific receptors involved for modulation of this form of sexual behaviour are briefly discussed. PMID- 3031700 TI - Physiologic responses to chronic dietary tyrosine supplementation in DOCA-salt treated rats. AB - The antihypertensive effect of chronic administration of L-tyrosine (Tyr) was investigated in a two-part study. In the first experiment, adult male Sprague Dawley rats were assigned to 1 of 4 treatment groups: control diet plus unilateral nephrectomy (Nphx) and 0.15 M NaCl (Sal) as the sole drinking solution (C-CTRL); control diet plus deoxycorticosterone acetate (DOCA, 268 micrograms/rat/day), Nphx, and Sal (C-DOCA); control diet supplemented with 2.5% L-p-Tyr plus Nphx and Sal (Tyr-CTRL), and Tyr plus DOCA, Nphx, and Sal (Tyr DOCA). Systolic blood pressure (SBP) increased within 2 weeks after initiation of treatment with DOCA-salt and remained elevated throughout the duration (8 weeks) of the study (p less than 0.001). Dietary administration of Tyr to DOCA-treated rats failed either to affect SBP in normotensive rats or the elevation of SBP in DOCA-treated rats. Dietary supplementation with Tyr induced a significant elevation in urinary excretion of free dopamine (week 1, 3, 5, and 7) and a decreased excretion of free norepinephrine (week 1) without regard to DOCA treatment. Metabolic responsiveness (change in colonic temperature) and cardiovascular responsiveness (change in heart rate) to subcutaneous administration of the beta-adrenergic agonist, isoproterenol, were significantly prolonged while alpha 2-adrenoceptor number (cerebral cortical membranes; 3H yohimbine binding) was reduced in rats receiving Tyr. In the second experiment, similar rats were assigned to 1 of 3 treatment groups: control diet plus Nphx and Sal, control diet plus Nphx, DOCA and Sal, and Tyr plus DOCA, Nphx, and Sal; however, Tyr was not started until DOCA-salt-induced hypertension developed (4 weeks). Neither acute (2.5 h post-meal) nor chronic (4 weeks) effects of administration of Tyr on SBP were noted. Thus, the Tyr-induced changes observed in these studies include a chronic increase in free dopamine, and a transient decrease in norepinephrine, excretion. No significant effects of Tyr on blood pressure of DOCA-salt-treated rats were observed. PMID- 3031701 TI - Actions of snake venom toxins on neuronal nicotinic receptors and other neuronal receptors. PMID- 3031702 TI - Identification of specific DNA lesions induced by three classes of chloroethylating agents: chloroethylnitrosoureas, chloroethylmethanesulfonates and chloroethylimidazotetrazines. PMID- 3031703 TI - Photoaffinity labeling of beta-adrenergic receptors. PMID- 3031704 TI - Photolysis of pheomelanin precursors: an ESR-spin trapping study. PMID- 3031705 TI - Photodynamic generation of hydroxyl radicals by hematoporphyrin derivative and light. PMID- 3031706 TI - The use of spin label oximetry in the study of photodynamic inactivation of Chinese hamster ovary cells. PMID- 3031707 TI - Is DNA topoisomerase involved in the UV excision repair process? New evidence from studies with DNA intercalating and non-intercalating antitumor agents. PMID- 3031708 TI - Laser flash photokinetic studies of rose bengal sensitized photodynamic interactions of nucleotides and DNA. PMID- 3031709 TI - Photoinduced generation of hydrogen peroxide and hydroxyl radicals in melanins. PMID- 3031710 TI - Spectroscopic studies of cutaneous photosensitizing agents--IX. A spin trapping study of the photolysis of amiodarone and desethylamiodarone. PMID- 3031711 TI - Restriction enzyme cleavage of UV-irradiated DNA: a comparison of far-UV and mid UV wavelengths. PMID- 3031712 TI - Free radicals detected by ESR from phenylheptatriyne in liposomes irradiated with UV-A. PMID- 3031713 TI - Light-flash physiology with synthetic photosensitive compounds. PMID- 3031714 TI - Influences of ACTH 4-10 on event-related potentials reflecting attention in man. AB - The present paper is concerned with effects of the 4-10 sequence of the endogenous ACTH on electrophysiological measures of attention in humans. It was attempted to replicate previous findings of an impaired selective attention following administration of an analog of ACTH 4-9. The effect of this analog had been found to dominate in the beginning of the blocks of an attention task, but to fade away with time on task. In the present study, fourteen male students were tested in a dichotic listening paradigm, 40 min after intranasal application of either 0.4 mg ACTH 4-10, or placebo. Averaged auditory evoked potentials (AEPs) to attended and inattended tone pips, EEG power spectra, heart rate and blood pressure, and behavioral performance were measured during task performance. ACTH 4-10 appeared to slightly impair selective attention as indicated by AEP responses. In particular, the positive shift of the AEP waveforms to inattended stimuli was reduced at the beginning of each block of tone pips under ACTH 4-10. The pattern of actions resembled the effects observed after administration of the more potent synthetic analog of ACTH 4-9 in the previous experiment. Effects of ACTH 4-10 on the AEPs to inattended stimuli, however, differed from influences of the synthetic analog in that they did not affect a rather wide latency range but concentrated on the latency range of the P200 component. PMID- 3031715 TI - Recognizing the adolescent drug abuser. AB - Adolescents are at high risk for using and abusing illicit drugs. Guidelines for recognizing drug abusers are presented as well as a staging process for progression of drug use. The family physician is in an ideal position to identify young users/abusers and to assist them and their families in obtaining much needed assistance. PMID- 3031717 TI - Effects of receptor blockers (methysergide, propranolol, phentolamine, yohimbine and prazosin) on desimipramine-induced pituitary hormone stimulation in humans- II. Prolactin. AB - In this report the effects of various receptor blockers on desimipramine (DMI) induced prolactin (PRL) secretion in healthy male subjects are presented. Each trial consisted of two administrations: one of DMI i.v. alone and one of DMI i.v. in combination with the respective receptor blocker: methysergide (serotonin (5 HT) receptor blocker), propranolol (beta receptor blocker), phentolamine (alpha 1/alpha-2 receptor blocker), yohimbine (alpha-2 greater than alpha-1 receptor blocker), and prazosin (alpha-1 receptor blocker). Following administration of methysergide (12 mg p.o., n = 12), a significantly lower (p less than 0.01) DMI induced PRL secretion compared to DMI alone in another group of subjects (n = 12) was observed. Combined administration with propranolol (15 mg i.v.) significantly enhanced the DMI-induced PRL secretion compared to DMI 50 mg i.v. alone (n = 18, incomplete block design) (p less than 0.01). Neither combined administration with phentolamine (60 mg i.v., n = 12), yohimbine (10 mg i.v., n = 6), nor prazosin (1 mg p.o., n = 12) significantly influenced the DMI-induced PRL secretion compared to DMI alone in the same subjects. The results of the present study, especially the inhibitory effect on DMI-induced PRL secretion of methysergide, indicate that the primarily noradrenaline (NA) and lesser serotonin (5-HT) reuptake inhibiting antidepressant DMI stimulates PRL secretion via 5-HT neurons. Furthermore, the significantly enhanced PRL release following combined administration of DMI and propranolol suggests that a noradrenergic inhibitory effect also may be involved in the transmission of the PRL stimulus. PMID- 3031716 TI - Effect of receptor blockers (methysergide, propranolol, phentolamine, yohimbine and prazosin) on desimipramine-induced pituitary hormone stimulation in humans- I. Growth hormone. AB - In this report the effects of various receptor blockers on desimipramine (DMI) induced growth hormone (GH) secretion in healthy male subjects are presented. Each trial consisted of two administrations: one of DMI i.v. alone and one of DMI i.v. in combination with the respective receptor blocker: methysergide (serotonin (5-HT) receptor blocker), propranolol (beta receptor blocker), phentolamine (alpha-1/alpha-2 receptor blocker), yohimbine (alpha-2 greater than alpha-1 receptor blocker), and prazosin (alpha-1 receptor blocker). DMI-induced GH stimulation was not significantly different after DMI i.v. alone (n = 12) than after three days' pretreatment with 12 mg methysergide p.o. in another group of subjects (n = 12). Following combined administration of DMI and propranolol (15 mg i.v.), GH secretion was significantly increased by 25 mg DMI (p less than 0.05) and 50 mg DMI (incomplete block design, n = 18). GH secretion was significantly lower (p less than 0.01) after DMI in combination with 60 mg phentolamine i.v. compared to that after administration of DMI alone in the same group (n = 12). Following 10 mg yohimbine i.v. in combination with DMI (n = 6), the DMI-induced GH increase was also significantly less (p less than 0.05) than that after DMI alone. The DMI-induced GH increase following DMI plus 1 mg prazosin p.o. (n = 12) was comparable to that after DMI alone. The results indicate that the GH-stimulating effect of DMI is primarily related to the ability of DMI to inhibit noradrenaline (NA) reuptake. Should serotonergic receptors be involved in the DMI-induced GH secretion at all, they transmit a positive stimulus. The alpha-1 receptors are most likely not (or not essentially) involved, whereas alpha-2 receptors affect the DMI-induced secretion positively, and beta receptors have an inhibitory effect. PMID- 3031718 TI - Effects of receptor blockers (methysergide, propranolol, phentolamine, yohimbine and prazosin) on desimipramine-induced pituitary hormone stimulation in humans- III. Hypothalamo-pituitary-adrenocortical axis. AB - In this report the effects of various receptor blockers on the desimipramine (DMI)-induced cortisol (ACTH) secretion in healthy male subjects are presented. Each trial consisted of two administrations: one of DMI i.v. alone and one of DMI i.v. in combination with the respective receptor blocker, i.e. methysergide (serotonin (5-HT) receptor blocker), propranolol (beta receptor blocker), phentolamine (alpha-1/alpha-2 receptor blocker), yohimbine (alpha-2 greater than alpha-1 receptor blocker), and prazosin (alpha-1 receptor blocker). In addition, the effect of prazosin on DMI-induced ACTH stimulation was examined. DMI-induced cortisol stimulation was not significantly different after DMI alone (n = 12) from that after three days pretreatment with methysergide (12 mg p.o.) in another group of subjects (n = 12). Neither the combination of DMI plus propranolol (15 mg i.v. n = 18, incomplete block design) nor that of DMI plus phentolamine (60 mg i.v. n = 12) had a significant influence on DMI-induced cortisol secretion. Following combined administration with yohimbine (10 mg i.v.), cortisol secretion was higher compared to that after DMI alone in the same group (n = 6). DMI induced cortisol secretion was significantly lower (p less than 0.01) following combined administration with prazosin (1 mg p.o. n = 12), as was DMI-induced ACTH secretion (p less than 0.05) in these subjects. The findings of these trials, especially those of the prazosin trial, indicate that DMI-induced stimulation of cortisol and ACTH secretion is attributable to the noradrenaline (NA) reuptake inhibiting effect of DMI, and that the stimulus is transmitted with the aid of noradrenergic alpha-1 receptors. Alpha-2 receptors possibly exert a negative influence on this effect. PMID- 3031720 TI - Membrane conductance and potassium permeability of the rat lens. AB - The membrane potential, electrical impedance and 86Rb+ efflux rate constants were measured in the rat lens perifused at 35 degrees C. The membrane conductance was obtained from the difference between the magnitude of the impedance at low (less than 0.1 Hz) and high (greater than 100 Hz) frequencies. Values of the rubidium permeability coefficients (PRb) were obtained from the rate constant and potential data. The values for the membrane potential and conductance in control solution (5 mM potassium) were -69.6 mV and 5.5 X 10(-4) S respectively, while the computed permeability was 2.9 X 10(-8) m.s-1. On perifusing with 35 mM potassium, the membrane depolarized by 25 mV and the conductance and rubidium permeability increased considerably. These increases could be blocked by quinine (0.3 mM), tetraethylammonium (30 mM) and 4-aminopyridine (10 mM). The latter agent was more effective at alkaline pH (8.3). It is suggested that there are voltage-gated potassium channels that are inhibited by these three agents. After the initial depolarization in high potassium, there was little further change in membrane potential with any of the inhibitors. All three agents, however, produced a marked depolarization when applied in control solution. This was accompanied by a decrease in conductance and rubidium permeability, suggesting that, in the rat lens, some voltage-gated potassium channels are activated at the resting potential. PMID- 3031719 TI - Environmentally-induced modification of the benzodiazepine/GABA receptor coupled chloride ionophore. AB - The benzodiazepine/GABA receptor coupled chloride ionophore was examined in brain membranes of rats maintained in either a conventional animal facility or a "protected" (low-stress) environment. Following a 10 min ambient temperature swim, animals maintained in both environments had qualitatively similar increases in the number (Bmax) of [35S]t-butylbicyclophosphorothionate (TBPS) binding sites, the apparent affinity of this radioligand, and the efficacy and potency of Cl- to enhance [3H]flunitrazepam binding. Nonetheless, the Bmax of [35S]TBPS and efficacy of Cl- to enhance [3H]flunitrazepam binding were significantly lower in animals housed in the protected environment compared to those maintained in a conventional facility both before and after swim stress. Furthermore, in rats housed in a protected environment, sequential removal of animals from a common cage (cohort removal), produced a very rapid increase (less than or equal to 15 s) in Cl(-)-enhanced [3H]flunitrazepam binding in cortical and hippocampal but not cerebellar membranes. Cohort removal also produced a sequential increase in the number of [35S]TBPS binding sites and apparent affinity of this radioligand in cerebral cortical membranes. The effects of cohort removal were not observed in animals subjected to ambient temperature swim or if animals were removed from different cages. Changes in the benzodiazepine/GABA receptor coupled chloride ionophore produced by cohort removal from a common cage preceded any statistically significant changes in circulating levels of alpha-MSH, beta endorphin, ACTH or corticosterone. These findings suggest that the benzodiazepine/GABA receptor chloride ionophore complex (supramolecular complex) is under both tonic and acute regulation by the environment, and may subserve a physiologically relevant function in the response to stressful or anxiety producing stimuli. PMID- 3031721 TI - Protection of mice against fission neutron irradiation by WR-2721 or WR-151327. AB - Two phosphorothioate compounds, WR-2721 and WR-151327, were examined for their radioprotective efficacies against the effects of fission neutron irradiation in male and female mice. Within sex groups no significant difference in lethality at 30 or 100 days postirradiation was found between WR-2721 or WR-151327 pretreatment. The dose modification factors (DMFs) for male mice treated with either compound were 1.29 (LD50/30) and 1.24 (LD50/100), and those for drug treated female mice were 1.21 (LD50/30) and 1.19 (LD50/100). Both WR-2721 and WR 151327 were found to be equally radioprotective when compared using DMFs as the end point. WR-151327 (500 mg/kg, ip) was found to be significantly more toxic to both male and female B6D2F1 mice than equimolar amounts of WR-2721. Small but significant sex differences in radioprotection were found: the DMFs for female mice pretreated with either compound were lower than those for similarly treated male mice; the incidence of mortality 31-100 days postexposure in male mice pretreated with WR-151327 was greater than for female mice. In addition, sex differences were noted in drug toxicity. Toxic death in female mice given WR 151327 (500 mg/kg, ip) is 2.6 times more probable than in males. PMID- 3031722 TI - The differential protection by WR2721 of skin versus growing cartilage following irradiation in weanling rats. AB - The potential for radioprotection of growing cartilage by the thiophosphate WR2721 was evaluated in weanling rats using single fractions of irradiation. Protection of acute skin toxicity was monitored simultaneously. Single doses of 600, 1200, 1800, or 2400 cGy were administered to the left tibia of CrL:CD(SD)BR female rats in groups of 12. Identically treated groups were injected with 310 mg/kg WR2721 (2/3 the determined LD50/30) in a concentration of 26 mg/ml intraperitoneally 15 min prior to irradiation. Rats untreated or given WR2721 without radiation served as control groups. Radiographs of the irradiated and unirradiated tibiae for each animal were obtained weekly to the date of sacrifice at 80 days following the initial treatment. Skin toxicity was assessed weekly starting on the second week using Moulder's scale (J.E. Moulder, J.J. Fischer, and A. Casey, Radiology 115, 465-470 (1975]. No significant difference in bone growth as measured by tibial lengths for the WR2721-treated or untreated animals was observed. Skin toxicity including moist desquamation occurred in irradiated limbs and was substantially less in rats treated with WR2721. As opposed to previous work with cysteamine, WR2721 as administered had no significant radioprotective effect on tibial growth in weanling rats but substantially reduced the accompanying skin toxicity. PMID- 3031724 TI - Hepatocellular carcinoma: treatment with intraarterial iodized oil with and without chemotherapeutic agents. AB - Thirty-one patients with hepatocellular carcinoma (HCC) were given either an intraarterial injection of iodized poppyseed oil (Lipiodol) alone (group A, n = 6), an emulsion of iodized oil and doxorubicin hydrochloride (Adriamycin) (group B, n = 15), or chemoembolization with the same emulsion followed by gelatin sponge (Gelfoam) particles (group C, n = 10). Hepatic resection was subsequently performed. The frequencies of complete necrosis of tumor in the main lesion, daughter tumors, tumor thrombus, and foci of intracapsular invasion were evaluated in the cut surface of the resected specimen. Group C demonstrated the best therapeutic effects, showing complete necrosis of the main lesion in 83% (P less than .01), daughter tumors in 53% (P less than .01), tumor thrombus in 17%, and foci of intracapsular invasion in 67%. These results are superior to those reported previously for chemoembolization without iodized oil. Group B showed better results than group A, but the difference was not significant. Iodized oil alone (group A) had practically no therapeutic effect but was helpful in differentiating small HCC from regenerative nodules. PMID- 3031725 TI - High-attenuation recent thrombus of the portal vein: CT demonstration and clinical significance. AB - Attenuation characteristics of portal vein thrombi on nonenhanced computed tomographic (CT) scans were assessed in 122 patients with proved portal vein thrombosis. Portal vein thrombi of high attenuation were found in four patients with hepatocellular carcinoma. From pathologic and radiologic studies, it was concluded that the high attenuation was caused by blood clots of recent onset formed at the tip of tumor thrombus. Differentiation from choledocholithiasis, hematobilia, and calcification of thrombi could be easily made by means of ultrasonography (US). Although plain CT is usually considered noncontributory in the diagnosis of venous thrombosis, it enabled the differentiation of recent thrombus in these four patients. Tumor thrombus in the major branches or main trunk of the portal vein is indicative of poor prognosis. When hepatic mass and high-attenuation portal vein thrombi are demonstrated with plain CT and substantiated by US, enhanced CT and angiography may be unnecessary for treatment of patients with advanced hepatocellular carcinoma. PMID- 3031723 TI - Cytomegalovirus infection of the alimentary canal: radiologic findings with pathologic correlation. AB - A spectrum of radiologic findings in cytomegalovirus (CMV) infection of the alimentary canal seen in 14 patients and correlated with pathologic examinations is described. Twelve patients had acquired immunodeficiency syndrome and two had no identified immunosuppression. Autopsies were performed on 12. Diffuse CMV colitis was present in eight patients, enteritis in seven, esophagitis in four, gastritis in two, cholangitis in one, and acute pancreatitis in one. Of 11 patients with enteritis and/or colitis seven had significant lower gastrointestinal bleeding and five died as a result of it. Radiologic findings in the gastrointestinal tract included superficial or deep mucosal ulcerations, perforation or fistula formation, luminal narrowing, rigidity and thickening of the intestinal wall, and inflammatory infiltration of the mesentery. These were seen on barium examinations and computed tomographic (CT) scans. Findings of pancreatitis were seen on CT scans in one patient. In another, a cholangiogram showed abnormal bile ducts caused by CMV cholangitis. The radiologist should be aware of the diverse manifestations of the disease and its likely occurrence in immunosuppressed individuals. PMID- 3031726 TI - Wilms tumor occurring as a botryoid renal pelvicalyceal mass. AB - Wilms tumor usually occurs as an abdominal mass arising from the renal parenchyma. A case was encountered in which the neoplasm filled the pelvicalyceal system of an 8-year-old boy as a botryoid mass, with minimal parenchymal involvement. The radiologic manifestations, pathologic features, and surgical implications are discussed. PMID- 3031728 TI - Light scan evaluation of nonpalpable breast lesions. AB - Transillumination light scanning of the breast was performed immediately before needle localization of 112 nonpalpable mammographic abnormalities detected in 103 patients. Twenty-four cancers were diagnosed in 23 patients. The positive predictive value for mammography was 21%. Ten (42%) of these nonpalpable cancers were not visible on transillumination light scanning. Eleven of 16 invasive ductal cancers and three of seven intraductal cancers were evident on transillumination light scans, but a single case of lobular carcinoma in situ was not evident. There were 12 false-positive light scan examinations. Transillumination light scanning had a 58% sensitivity, an 86% specificity, a 54% positive predictive value, and an 88% negative predictive value for small breast lesions. Therefore, the authors are unable to recommend delaying biopsy in patients with mammographic abnormalities on the basis of a negative light scan study. PMID- 3031727 TI - Benign extraaxial tumors: contrast enhancement with Gd-DTPA. AB - Meningiomas, acoustic neuromas, and other benign extraaxial tumors have little contrast with adjacent brain tissue on conventional magnetic resonance (MR) images. The contrast enhancement produced by intravenous administration of 0.1 mmol/kg of gadolinium-DTPA in these tumors was measured on T1 MR images. Acoustic neuromas showed the greatest enhancement (average, 310%), meningiomas the next greatest (average, 180%), and neurofibromas, glomus tumors, and pituitary microadenomas the least enhancement. The degree of enhancement was almost always greater at 3 minutes than at 25 or 55 minutes. Contrast between the tumor and adjacent tissue resulted from tumor enhancement in neuromas, meningiomas, and neurofibromas and from enhancement of the surrounding tissue in pituitary microadenomas. PMID- 3031729 TI - Identification of ACTH-producing intrathoracic tumors by measuring ACTH levels in aspirated specimens. AB - Three patients with Cushing syndrome secondary to ectopic adrenocorticotropic hormone (ACTH) production underwent direct 22-gauge needle aspiration of bronchial (two cases) and mediastinal (one case) carcinoid tumors. High levels of ACTH were measured in all three tumors. This technique permits absolute identification of an ectopic source of ACTH before surgery. PMID- 3031730 TI - Aspects on the information handling by the central nervous system: focus on cotransmission in the aged rat brain. PMID- 3031731 TI - Morphofunctional studies on the neuropeptide Y/adrenaline costoring terminal systems in the dorsal cardiovascular region of the medulla oblongata. Focus on receptor-receptor interactions in cotransmission. PMID- 3031732 TI - Characterization of central neuropeptide Y receptor binding sites and possible interactions with alpha 2-adrenoceptors. PMID- 3031733 TI - Neuropeptide Y receptor interaction with beta-adrenoceptor coupling to adenylate cyclase. PMID- 3031734 TI - Cotransmission at GABAergic synapses. PMID- 3031735 TI - Transcellular metabolism of eicosanoids. AB - In this chapter, current concepts of eicosanoid biochemistry and function have been summarized. Emphasis has been placed on the importance of ascertaining a complete profile of eicosanoids synthesized by a given tissue under varying conditions of stimulation. The concept of transient intermediates, which are involved in each of the known pathways, was reviewed. Such intermediates can also represent substrate for cell-cell interactions, which have been classified and discussed. Knowledge of the synthetic mechanisms, metabolism, and catabolism of eicosanoids has surpassed our comprehension of their biologic actions. It can be speculated that eicosanoids reinforce or synergize normal homeostatic mechanisms that might proceed less effectively in their absence but occur more efficiently when they are produced. Incomplete comprehension of the functional role of eicosanoids has retarded the development of definitive pharmaceutical approaches toward inhibition or stimulation of eicosanoid production in pathophysiologic situations. With regard to thrombosis, the inflammatory response and host-defense mechanisms, leukotrienes, which have been shown to act on vascular endothelium and smooth muscle, may play an important role in occlusive vascular disease. Since leukotrienes and other hydroxy acids such as 12-HETE are not inhibited by aspirin or nonsteroidal anti-inflammatory agents, development of a unifying concept is difficult. It appears that we are approaching a point in time for reevaluation and reassessment of the role of the eicosanoid pathway in hemostasis and thrombosis. Most importantly, recent eicosanoid research has provided answers to biologic phenomena that were not explicable in the past and also explains why therapeutic trials in occlusive vascular disease were not as successful as predicted on theoretical grounds. PMID- 3031736 TI - Inositol phospholipid turnover in stimulus-response coupling. PMID- 3031739 TI - [Transmission of cellular information]. PMID- 3031737 TI - The role of specific forms of heparan sulfate in regulating blood vessel wall function. PMID- 3031738 TI - Hereditary disorders predisposing to thrombosis. PMID- 3031740 TI - [recA protein]. PMID- 3031741 TI - Neutron therapy for malignant tumours of the salivary glands. A report of the Edinburgh experience. AB - A group of 28 patients with malignant tumours of the salivary glands have been treated by d(15) + Be neutron irradiation. Nineteen patients had inoperable cancers. Three had gross recurrent cancer and three had measurable residual cancer after surgery. Three patients were treated post-operatively for microscopic residual disease. Seven different histological types of tumour were included. Six out of 8 patients with adenoid cystic carcinomas have lasting local tumour control. 54.5% of the gross tumours were locally controlled. All three of those classified as microscopic residual disease have no evidence of local recurrence. 11/14 cancers given 16.0 Gy or more in 20 fractions in 4 weeks were controlled compared with only 1/8 given a lower dose. 12/19 cancers less than 10.0 cm maximum diameter were controlled. The radiation-related morbidity was similar to that observed after photon therapy. PMID- 3031742 TI - Chromogranin A, B and C: immunohistochemical localization in ovine pituitary and the relationship with hormone-containing cells. AB - Chromogranins A, B and C, three distinct groups of proteins found in bovine chromaffin granules, were also found to be present in the pituitary using immunoblotting techniques. Their distribution was therefore studied in the normal ram pituitary using an immunoperoxidase technique applied to semithin serial sections and compared with that of some of the hormones of the anterior pituitary. Chromogranin-immunoreactivity was found in gonadotrophs (all three), thyrotrophs (A with some positive for C) and corticotrophs (a fraction with A and fewer with B and C). The mammotrophs and somatotrophs were negative. Chromogranin C was the only one of the three to be located in the pars nervosa, whilst chromogranin B was rarely found in the pars intermedia. The results suggest that chromogranins A, B and C are not always stored together, some hormone-containing cells do not contain immunohistologically detectable levels of the chromogranins. PMID- 3031743 TI - Opioid inhibition of immunoreactive corticotropin-releasing factor secretion into the hypophysial-portal circulation of rats. AB - The role of the opioid peptides in the regulation of adrenocorticotropin (ACTH) secretion remains unclear. In rats, morphine and the enkephalins exert a stimulatory effect on the hypothalamic-pituitary-adrenal axis, while beta endorphin (beta-E) and dynorphin (DYN) are reported to have stimulatory or inhibitory activity. Alternatively, data from human studies indicate a clear inhibitory role of opiates. In the present studies, secretion of immunoreactive corticotropin releasing factor (irCRF) into the hypophysial-portal circulation was directly measured before and after intracerebroventricular administration of beta-E, DYN and naltrexone (NTX). Both beta-E and DYN were equipotent in their dose-related inhibition of irCRF secretion. The inhibitory action of beta-E was reversed by NTX, while the action of DYN was only partially blocked. Administration of NTX alone resulted in a significant elevation of spontaneous and stimulated irCRF secretion. Finally, injection (icv) of 1.0 nmol beta-E or DYN blocked the nitroprusside-hypotension induced elevation of irCRF. These observations suggest, that under the conditions of these experiments, exogenous beta-E acting primarily via mu opioid receptors and DYN acting via kappa and mu receptors exert tonic inhibitory effects on the activity of CRF secreting cells. Furthermore, it appears that beta-E and DYN are capable of modulating (inhibiting) stimulated secretion of irCRF and thus activity of the hypothalamic pituitary-adrenal axis. PMID- 3031744 TI - Alcohol reverses the proconflict effect of corticotropin-releasing factor. AB - Alcohol has tension reducing properties in man that are reflected in a release of punished responding in a rat operant conflict test. In contrast, corticotropin releasing factor (CRF), injected centrally produces a suppression of punished and non-punished responding in the conflict test consistent with its hypothesized role in mediating behavioral responses to stress. Alcohol in a dose of 0.75 g/kg reversed the suppressive effects of 0.5 microgram CRF injected intracerebroventricularly on punished responding but augmented the suppression of unpunished responding by CRF. Results suggest that one mechanism for the tension reducing properties of acute alcohol intoxication may involve a suppression of brain CRF systems. PMID- 3031745 TI - Corticotropin-releasing factor receptor antagonist: effects on the autonomic nervous system and cardiovascular function. AB - The corticotropin-releasing factor (CRF) receptor antagonist, alpha-helical [Glu27]-corticotropin-releasing factor 9-41 (CRF 9-41) has been assessed for its ability to modify plasma concentrations of epinephrine and norepinephrine, mean arterial pressure (MAP) and heart rate (HR). Basal concentrations of epinephrine and norepinephrine were not altered by lateral ventricular (icv) administration of CRF 9-41. However, this CRF antagonist, given icv, attenuated the rise of plasma epinephrine following 30% hemorrhage and insulin-induced hypoglycemia. CRF 9-41 did not alter the increased plasma concentrations of epinephrine or norepinephrine following icv administration of bombesin. Icv administration of CRF 9-41 blunted CRF-induced elevation of MAP and HR in normal animals. However, this CRF antagonist did not modify the MAP or HR in spontaneously hypertensive rats. Similarly, this CRF antagonist administered to Sprague-Dawley rats neither prevented the rise of MAP or HR following electrical stimulation of the central nucleus of the amygdala, nor did it affect nitroprusside-induced hypotension and tachycardia. PMID- 3031746 TI - Atrial natriuretic peptide does not affect corticotropin-releasing factor-, arginine vasopressin- and angiotensin II-induced adrenocorticotropic hormone release in vivo or in vitro. AB - The effect of synthetic atrial natriuretic peptide (ANP) was examined on the in vivo and in vitro release of ACTH. Intravenous ANP (4 micrograms/kg body weight) administration did not affect the corticotropin releasing factor (CRF, 4 micrograms/kg body weight)-, arginine vasopressin (AVP, 2 micrograms/kg body weight)- and angiotensin II (A II, 4 micrograms/kg body weight)-induced ACTH release in unanesthetized freely moving rats. ANP did not inhibit the basal, CRF- and AVP-induced release of ACTH in pituitary cell cultures. ANP did not affect the CRF- and AVP-induced plasma corticosterone elevation, while it attenuated the AVP-induced corticosterone elevation. These results indicate that ANP does not affect the ACTH release at the pituitary level in vivo and in vitro. PMID- 3031747 TI - [The use of hydroxyapatite in cases with endodontic and parodontic lesions]. PMID- 3031749 TI - [Retrovirus, oncogenes and malignant transformation]. PMID- 3031748 TI - [Biological control of Culicidae and Simuliidae: bacterial insecticides]. PMID- 3031750 TI - [Jaundice of 1-month's development and cerebrovascular accident in a 66-year-old male]. PMID- 3031751 TI - [Cardiac MRT. Research technic and topographic anatomical imaging information]. AB - ECG-triggered cardio-MRT, using T1-weighted SE sequences, provides images of the heart showing very high anatomical resolution. So far, however, it has not been possible to demonstrate the morphology of the valves and coronary vessels. By using multiplanar sections, it is possible to obtain cuts which are perpendicular to the portion of myocardium or valve under consideration. This provides optimal imaging for morphological diagnosis and for evaluating the ventricular myocardium and its function. PMID- 3031752 TI - [Percutaneous balloon dilatation of pulmonary valve stenosis]. AB - Balloon valvuloplasty was carried out on five children with isolated valvular pulmonary stenosis. One child had a residual stenosis following a Brock's transventricular valvotomy. The systolic gradient between the right ventricle and the pulmonary artery fell from 60.4 +/- 18.7 mmHg to 21.8 +/- 12.0 mmHg. Right ventricular pressure fell from 76.2 +/- 20.7 mmHg to 43.0 +/- 16.4 mmHg. Percutaneous balloon valvuloplasty has proved to be an effective method for the treatment of isolated valvular pulmonary stenosis. PMID- 3031754 TI - [Computed tomographic diagnosis and differential diagnosis of leiomyosarcoma of the inferior vena cava]. AB - The computer tomographic appearances of the rare leiomyosarcoma of the inferior vena cava are described and illustrated by one case. CT is particularly informative and the differential diagnosis from caval thrombosis and extension of tumour thrombus into the cava is discussed. PMID- 3031753 TI - [Direct venous thrombolysis and venous angioplasty in the upper extremity]. AB - Venous thromboses of stenoses in the upper extremity are often the result of a compression syndrome of the shoulder girdle, the Paget-von Schroetter syndrome, vascular surgery, space-occupying lesions in the mediastinum or the result of catheterisation. Direct venous thrombolysis and venous angioplasty were performed successfully in six patients. PMID- 3031755 TI - [Magnetic resonance tomography (MRT) of the orbit]. AB - By using high field strengths and surface coils, MRT achieves a resolution comparable with CT in the orbita. The advantages of MRT are good contrast resolution and imaging in several planes. Twenty-six patients have been examined by MRT, which has shown high sensitivity and good detail for the demonstration of pathological changes. In spite of this, MRT at present is not a realistic alternative to ultrasound and CT, because it is unable to demonstrate bone and calcification; its specificity is low, but the time and cost of the examination is high. It is indicated only for problems involving the optic nerve and chiasma. PMID- 3031756 TI - [Malignant tumors of the mouth and pharynx--MR tomography with surface coils]. AB - The place of MRT, using surface coils, has been evaluated retrospectively in 27 patients with clinically confirmed malignant tumours of the mouth and pharynx. The extent of the tumour and its relationship to neighbouring structures is well demonstrated. The resolution obtainable by using surface coils is almost as good as that of CT. Soft tissue contrast is better in showing the spread and size of the tumour. A disadvantage of MRT is the inability in most cases to demonstrate bone destruction. Differentiation between residual tumour, recurrence and scarring is possible only occasionally. Quantitative MRT may be expected to lead to improved diagnosis during the course of following up. PMID- 3031757 TI - [Functional echo morphology of Bauhin's valve]. AB - Following rectal irrigation, it is possible to demonstrate the ileo-caecal valve sonographically, using a right para-inguinal approach. The shape, position and motility of the valve can be evaluated. The normal valve is two to three mm. thick, similar to the colonic wall, but its lips are thickened. Two distinct mechanisms are responsible for opening the valve, the ileocolic pressure gradient and relaxation of the circular muscle round the valve. The resulting intraluminal flow can be quantified by duplex sonography. The information may be of diagnostic and therapeutic significance, particularly during childhood. PMID- 3031758 TI - [Differential diagnosis of chronic peripancreatic pseudocysts on the computerized tomogram. Possibility of error and pitfalls]. AB - Cystic masses within the pancreas and in its neighbourhood are often really pseudocysts. In most cases, centrally necrotic solid tumours or genuine cystic neoplasms are easy to differentiate from such pseudocysts. They are in fact rare pathologic conditions. The exclusion of a pseudocyst is sometimes more difficult, especially in peripancreatic cystic masses. Computed tomography has significantly improved the diagnostics of the upper abdomen. Nevertheless, it sometimes creates new problems. This study is a selection of cases pointing to differential diagnostic difficulties that may become pitfalls for the examiner. An image imminent approximative diagnosis is attempted. The importance of the localisation of these lesions and of the topographical anatomy of the upper abdomen are pointed out. PMID- 3031759 TI - [Intra-arterial DSA versus conventional angiography in assessing the resectability of pancreatic and periampullary carcinomas]. AB - Fifty patients were examined to determine whether intraarterial DSA can replace conventional angiography to assess the resectability of pancreatic and periampullary carcinomas. Intraarterial DSA yielded equivalent diagnostic information in 60% of the cases. As a result of contrast-enhanced visualisation of the splanchnic veins intraarterial DSA was superior to conventional angiography in 26% of the cases, whereas in 14% intraarterial DSA provided less information due to motion artifacts. PMID- 3031760 TI - [MR of the shoulder joint with surface coils at 1.5 tesla. Its anatomy and possible clinical use]. AB - High spatial resolution magnetic resonance images of the shoulder were obtained in axial, sagittal and coronal orientations using a 1.5 T imaging system and anatomically shaped, wrap-around surface coils. Variations in scapular position induced by patient positioning change the relationship of the planes to the shoulder anatomy and make reproducibility of sagittal and coronal planes difficult. We, therefore, use--after axial orientation--image-oblique planes perpendicular and parallel to the glenoid fossa. In this manner MRI can visualise the anatomic structures of the shoulder including rotator cuff, long biceps tendon, articular capsule, articular cartilage, muscles and bones due to the high soft tissue contrast of MRI. PMID- 3031761 TI - [Results of bone marrow scintigraphy in hematological diseases]. AB - Bone marrow scintigraphy was performed in 40 patients with haematological disorders (lymphomas, multiple myeloma, myeloid leukaemia, panmyelopathy and others). Routine skeletal scintigraphy was available for comparison in 18 patients. In agreement with the literature it became evident that marrow scintigraphy can show infiltrations of the bone marrow in haematological disorders earlier than skeletal scintigraphy. Hot lesions were caused by reactive proliferation of the bone marrow such as may occur with acute inflammatory joint diseases, osteomyelitis, fracture healing or other entities. Marrow imaging demonstrated reliably the actual distribution of the functioning bone marrow, a characteristic that is important for diagnosis and staging, especially in myeloproliferative diseases. PMID- 3031762 TI - [Microangiographic research on revascularization in the fracture model of the canine tibia]. AB - Micro-angiograms were performed between eight weeks and one year following osteotomies and pressure osteosyntheses in 34 dog tibiae. Seventeen animals also underwent lumbar sympathectomy. In more than half of the cases, vascularisation of the medulla was reduced by damage to the nutrient artery. Segmental connections could be demonstrated by means of transcortical anastomoses. Following sympathectomy, revascularisation was more marked as a result of numerous trans-osseus anastomoses. This, however, did not lead to improved healing; healing was worse after sympathectomy. Transcortical revascularisation and vessel growth along drill holes are of crucial significance. PMID- 3031763 TI - Lymphography and abdominal computed tomography in staging Hodgkin's disease. AB - Lymphography and abdominal CT were performed in 78 patients staged for Hodgkin's disease. In 82% of all patients, both examinations agreed on the presence or absence of lymph node involvement. In the group of 39 patients undergoing lymphography prior to CT, the agreement was 90%. In the group of 39 patients with lymphography following CT, the agreement was 74%. In 50% of the patients with discrepant findings, lymphography revealed abnormal nodes compared with CT. Lymphography was abnormal in 26% of patients with a normal CT scan as the first examination, and in 9% of patients with a normal CT scan as the second examination. It is concluded that lymphography is more reliable than CT in the examination of the abdomen. CT performed after a normal or an abnormal lymphogram adds little additional information. When CT is preferred as the initial investigation in staging Hodgkin's disease, lymphography only adds significant information if the CT scan is normal or equivocal. PMID- 3031765 TI - [The importance of thermography in the diagnosis of lumbar radicular pain syndromes]. AB - This study proved the existence of lateral differences in thermographic recordings of the legs of 83 out of a total of 84 patients with the clinical diagnosis "lumbar disk herniation". In 68.5 per cent only of 71 patients with unilateral symptoms, however, the hypothermic areas occurred in the symptomatic leg. These areas could not be matched with corresponding dermatomas. As regards determination of the level of the herniation, thermography is an unreliable method for differentiation between the different segments, because the site of the dermatoma cannot be identified in relation to the site of the herniation. The mechanism of difference in temperature distribution probably depends on sympathetic reflexes with increased activity in the vasoconstrictor system due to pain. These reflexes are not limited to one dermatoma. This has been confirmed by means of diagnostic nerve blocks of the appropriate root. PMID- 3031764 TI - [Diagnosis of muscle hematomas and pseudotumors in hemophilia]. AB - Two hundred and twenty-six patients with haemophilia A and B were examined by CT. In 135 cases, some form of bleeding was demonstrated. CT permits exact anatomical localisation of the haematoma and its delineation from neighbouring structures. Measurements of the densities of the haematomas make it possible to determine their age from the change in absorption values. Computed tomography makes it easier for the clinician to choose between conservative or surgical treatment and to determine the duration of treatment during the regression of the haematoma. PMID- 3031766 TI - [Lumbar reprolapse or scar tissue? An attempt at differentiation via a computer tomographic bicolor mode]. AB - The bicolour mode for computer tomography is explained. It replaces the soft tissue level of the computer tomogram by setting the window level to +55 Hu and its width to 1 Hu. Structures in the spinal canal with attenuation values greater than 56 Hu appear white. The cauda equina has an attenuation value of less than +45 Hu. Disc prolapses, scar tissue and solid tumours (with the exception of lipomas) have attenuation values greater than +55 Hu. The use of the bicolour mode combined with intravenous contrast in patients with post-operative symptoms can often help to differentiate between reprolapse and scarring. A time-saving method for the bicolour mode is described. PMID- 3031767 TI - [Tinnitus and its radiological diagnosis and therapy]. AB - Tinnitus is a familiar symptom in diseases of the central nervous system. Its aetiology being of a varied nature, the type of tinnitus is a pointer to the pathoanatomic findings that are responsible for the disease. The causes of tinnitus are often found in the borderline areas between various neighbouring disciplines, such as otology, neurology and neurosurgery, whereas the final identification of the real causes is mainly within the scope of radiological diagnosis, in which computed tomography and superselective angiography play an essential role. In addition to arteriovenous fistulas near the petrous bone and glomus tumours, which are well known, there are a few other aetiologies of pulse synchronous tinnitus that are being discussed in this paper. In recent years there has been substantial therapeutical progress owing to the introduction of new techniques and the development of improved materials for embolisation in interventional neuroradiology. Their application is discussed in connection with various patient groups. PMID- 3031768 TI - [Parametric visualization of cerebral perfusion by intravenous digital angiography]. AB - Intravenous digital subtraction angiography is often used for evaluation of the extracranial and major intracranial vessels. The information obtained by this examination concerning cerebral perfusion is usually judged visually. In 50 patients a time-density curve at a specific point was analysed using 128 X 128 matrix. The level of grey scale at the moment of arrival of the bolus permits estimation of the haemodynamics of the intracranial vessels and provides a simple demonstration of differences of perfusion. PMID- 3031769 TI - [Contrast medium reaction to iohexol (Omnipaque) with monocyte activation demonstrated in vivo]. AB - A contrast reaction to the intravenous injection of the non-ionic contrast medium iohexol (Omnipaque) is reported. The symptoms developed several hours after the injection and at first were not recognised as being due to the contrast. The reaction could be reproduced under controlled conditions and was associated with a significant rise in thromboplastin synthesis in circulating monocytes, a sign of marked cellular activation. PMID- 3031770 TI - [Definitions of pulse sequences and parameter weightings in the NMR tomogram]. AB - Due to the different possibilities of image creation in MR tomography there is no clear terminology of pulse sequences and MR images. This paper tries to define the designation of pulse sequence parameters in a practical way and to specify the term "parameter weighting of MR images". Starting with general definitions, special definitions for CNS and liver are elaborated. PMID- 3031771 TI - [Cerebral manifestation of Nocardia brasiliensis infection]. PMID- 3031772 TI - CT-appearance of cervical actinomycosis. Case report. PMID- 3031773 TI - [Ossifying soft tissue tumor in melorheostosis]. PMID- 3031774 TI - [Idiopathic Ceelen-Gellerstedt pulmonary hemosiderosis]. PMID- 3031775 TI - Small renal haemangioma diagnosed with pharmacoangiography. A case report. PMID- 3031776 TI - [Rare complication of elbow joint arthrography]. PMID- 3031777 TI - [Comparative experimental pharmacodynamic study of different neuromuscular blockers]. PMID- 3031778 TI - [Bone metastases as the initial manifestation of hepatocarcinoma]. PMID- 3031779 TI - [Puncture aspiration with fine needle under echographic control. Initial study]. PMID- 3031780 TI - [Mixed hepatoblastoma in an adult]. PMID- 3031781 TI - [Primary herpes infection of the esophagus]. PMID- 3031782 TI - [The incidence of generalized metastases in hepatoma: apropos of a case]. PMID- 3031783 TI - [Determination of the distribution of ventilation-perfusion ratios: technic of elimination of multiple inert gases]. AB - The multiple inert gas elimination technique (MIGT) facilitates the estimation of the distributions of ventilation-perfusion (VA/Q) ratios in the experimental and clinical setting. The most relevant technical aspects and equipment and operational requirements needed to measure a mixture of inert gases in both the gas phase and the blood phase using gas chromatography are overviewed with detail. Results obtained in 3 dogs and 4 syringe-homogeneous lung models were entirely consistent with data formerly reported in the literature. Particular attention is paid to the linearity of the gas chromatograph detectors, reproducibility of inert gases sampling, and analysis of brands of heparin to detect acetone content. The errors of measurement (coefficients of variation) in blood were: 1.4 for sulfur hexafluoride; 1.8% for ethane; 2% for cyclopropane and halothane, each; 2.4% for diethyl ether; and, 3.6% for acetone. Important practical points are also emphasized in order to draw attention to potential problems and issues that should be concentrated upon to minimize the error in the measurements. It is concluded that the setting up of the MIGT is well established and validated. PMID- 3031784 TI - [Effect of the amplitude and frequency of sinusoidal stimulation on the harmonic distortion of the response of sensory terminations of muscle spindles]. AB - Harmonic distortion (HD) from 1,055 responses of muscle spindles sensory endings to sinusoidal stretches (frequency range 0.0008 to 0.8333 Hz, amplitude range 0.019 to 3.09 mm) has been studied in the cat soleus muscle. Sixty-six per cent were primary afferents (Ia) and 34% secondary (II). HD mean value (0.28) did not show any significant differences between both types of endings. Analysis of variance for HD versus stimulation amplitude showed a greater HD when stretch amplitudes were beyond 1.599 mm or less than 0.031 mm on primary afferents (p less than 0.001) and less than 0.070 mm on secondary (p less than 0.001). The effect of stimulus frequency was also significant (p less than 0.01 Ia and p less than 0.001 II), however only at 0.8333 Hz and in secondary endings HD was significantly higher. The silent period in the response, at release of stretch, caused by half wave rectification could explain about 50% of measured HD. PMID- 3031785 TI - Effect of fenoterol on secretions of an isolated single submucosal gland from the trachea. AB - The mechanism of secretion of airway submucosal glands is not known. Although the existence of cyclic AMP in the submucosal gland has been reported, the effect of stimulation of the gland by exogenous cyclic AMP has been denied. We hypothesize that the negative effect of exogenous cyclic AMP is due to the modified influence of ciliated epithelium on the submucosal gland, because epithelium releases chemical mediators. We isolated a single submucosal gland from a cat trachea. We measured mucous glycoprotein labeled with [3H]-glucosamine from the isolated submucosal glands. With stimulation of dibutyryl cyclic AMP, a percent increase of the secretion above control was about 3 times in isolated glands. Methacholine and fenoterol secreted mucus largely in isolated glands. We suggest that cyclic AMP increased the secretion in the glands. The difference of quantity of secretions between methacholine and fenoterol has been investigated further. PMID- 3031786 TI - Adrenergic and non-adrenergic, non-cholinergic control of airways. AB - In addition to cholinergic neural mechanisms, airway tone is influenced by adrenergic mechanisms and by more recently described neural mechanisms which are non-adrenergic and non-cholinergic (NANC). Sympathetic innervation to human airways is very sparse and there is no functional adrenergic innervation of smooth muscle, although sympathetic fibres may supply ganglia, submucosal glands and bronchial vessels. Airway tone may be influenced by circulating adrenaline and there is some evidence that adrenaline secretion may be impaired in asthma. beta-Adrenoceptors (which are almost entirely of the beta 2-subtype) are localized to many cell types in airways and beta-agonist may be beneficial in airway obstruction, not only by directly relaxing airway smooth muscle (from trachea to terminal bronchioles), but by inhibiting mast cell mediator release, by modulating cholinergic nerves, by reducing bronchial oedema and by reversing the defect in mucociliary clearance. There is little evidence that beta-receptor function is impaired in asthma. Alpha-adrenoceptors, which are bronchoconstrictor, may be activated by inflammatory mediators and disease, and alpha-agonists cause bronchoconstriction in asthmatic patients. However, alpha antagonists have little effect, which questions the role of alpha-receptors in asthma. NANC nerves which relax human airways have been demonstrated in vitro. Although the neurotransmitter is not certain, there is now convincing evidence that it may be vasoactive intestinal peptide (VIP) and a related peptide histidine methionine (PHM). VIP and PHM immuno-active nerves are found in human airways, and both peptides potently relay human airways in vitro (but not in vivo because of diffusion and metabolism problems).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3031787 TI - [Positron emission tomography of lung cancer using 18F-2-fluoro-2-deoxy-D-glucose and L-[11C-methyl]-methionine]. PMID- 3031788 TI - Markers of activity of sarcoidosis. PMID- 3031789 TI - Angiotensin-converting enzyme activity in sarcoidosis and other disorders. AB - A review is given on S-angiotensin-converting enzyme (SACE) and its clinical value, based upon 327 sarcoidosis patients and 1,274 patients with various disorders. SACE was elevated in 55% of the sarcoidosis patients, although with a higher frequency in those with active disease. Erythema nodosum was associated with normal initial SACE, subsequently rising, and sarcoid hypercalcaemia was consistently followed by elevated SACE. In non-sarcoid patients, elevated SACE was observed in only 10 cases. The sensitivity and specificity were 0.55 and 0.99, respectively, and the positive and negative predictive values were 0.95 and 0.90, respectively. Elevated SACE pointed strongly towards the presence of sarcoidosis, although reservations must be made in patients with liver disorders, diabetes mellitus, hyperthyroidism, asbestosis or silicosis which are rather common disorders also associated with elevated SACE. Normal SACE does not exclude sarcoidosis. PMID- 3031790 TI - Azathioprine treatment of chronic pulmonary sarcoidosis. AB - The object of this study was to evaluate the effectiveness of the immuno suppressor Azathioprine (AZ) on chronic and severe pulmonary sarcoidosis, with a persistent activity resistant to prolonged corticotherapy. The study was done on 10 patients (4 women, 6 men) afflicted by an histologically proven and chronic sarcoidosis, resistant to steroid treatment. The treatment consisted of a daily oral intake of 150 mg of AZ for six months. Its effectiveness was evaluated before and after treatment, in comparison with a control group and a steroid treated group of sarcoid patients. Serologic and alveolar functional and immuno biologic tests were performed in 8 cases according to the activity criteria defined at the IXth International Congress on sarcoidosis. No clinical or hematological side effects were observed; a clear and prolonged radiological and clinical amelioration was observed in 7 out of 10 cases and in 3 cases a restoration of sensibility to the tuberculin skin test; in the 8 cases a significant improvement (p less than 0.01) was noted after the sixth month of treatment only in the alveolar fluid in the following parameters: ACE, all the proteins studied and the percentage of lymphocytes. PMID- 3031791 TI - Neurosarcoidosis. A report of ten patients illustrating some usual and unusual manifestations. AB - The common and not so common neurologic lesions due to sarcoidosis include cranial nerve palsies, meningitis, hypothalamic and pituitary lesions, basilar meningitis, polyneuropathy, space occupying lesions and spinal cord involvement. The diagnosis of neurosarcoidosis requires a compatible clinical or radiologic picture of sarcoidosis and histological confirmation of noncaseating granulomas. Serum ACE, Gallium-67 lung scan and bronchoalveolar lavage are of help in supporting the diagnosis and establishing activity of the disease. A CT scan with enhancement is the diagnostic procedure of choice. Corticosteroids are the cornerstone of therapy. PMID- 3031792 TI - [Epidemiological study of otorhinolaryngologic cancers in Abidjan (Ivory Coast). Apropos of 204 cases]. PMID- 3031793 TI - Radiolabeled antibody therapy. PMID- 3031794 TI - Considerations with cisplatin (Platinol) administration in the office setting for oncology nurses. PMID- 3031795 TI - [Radiculo-spinal discomfort and Scheuermann's disease. 12 cases]. AB - Twelve cases of Scheuermann's disease associated with neurological disorders are reported. The relation between the two conditions can be established through herniated disc, kyphotic angulation or spinal cyst. A more interventionist attitude is justified by the progress and results of neuroradiological investigations and surgical techniques. PMID- 3031796 TI - Peripheral and spinal inputs to physiologically identified thalamic and nonthalamic relay neurons in cat cuneate nucleus. AB - A single-unit population study of the feline cuneate nucleus was carried out to identify principal neuron types, their distribution within the nucleus, pattern of peripheral activation, and receptive field characteristics. Units were also tested for response to isolated dorsal column or dorsolateral funicular electrical stimulation. The nucleus was explored in a uniform pattern, and sample size was optimized by applying the search stimulus shocks to the dorsal spinal cord. Single units were defined as spinal afferents, cuneothalamic-relay (CTR) neurons, and non-cuneothalamic-relay (non-CTR) neurons. The following features were observed: The distribution within the nucleus of specific cell types agreed with cytoarchitectural studies: Spinal afferent fibers were superficial and caudal; 22% of neurons were CTR neurons; CTR neurons were most dense in the middle of the nucleus and were largely separate from non-CTR neurons. Of the 58 neurons tested for response to isolated dorsal column and dorsolateral funicular stimulation, 24% were activated from both tracts. Convergent input from the off focus periphery (defined as other than the ipsilateral forelimb) was detected in both CTR and non-CTR neurons, most commonly from the contralateral forepaw. Several neurons were activated from three limbs. Thirty-seven percent of units were unresponsive to hair movement, touch, muscle palpation, or movement of joints. Compared to spinal fibers and non-CTR neurons, CTR neurons were most likely to have an identifiable input. PMID- 3031797 TI - [Management and long-term therapy of patients following aortocoronary bypass surgery, PTCA and thrombolysis treatment]. PMID- 3031799 TI - [Calcium antagonists--a new class of drugs used in the treatment of lung diseases (current status and outlook)]. PMID- 3031798 TI - [2 cases of stiff-man syndrome with tremor]. AB - Clinical history, neurologic investigations, course, EMG findings, muscle biopsy and response to diazepam treatment in two patients fulfilled criteria established by Gordon, Januszko and Kaufman for the diagnosis of the stiff-man syndrome. Both patients also presented tremor, a rare finding in this affection. PMID- 3031800 TI - [Current pathogenetic aspects in viral infections of the respiratory system]. PMID- 3031801 TI - [The current situation with intestinal tuberculosis]. PMID- 3031802 TI - [Importance and efficiency of an epidemiological survey of children with tuberculosis]. PMID- 3031803 TI - [Disease of the small airways as a posttuberculous syndrome]. PMID- 3031804 TI - [Value of determining the total E-rosettes in the pleural fluid]. PMID- 3031805 TI - [Pulmonary dysfunction study of children with chronic respiratory symptoms]. PMID- 3031806 TI - [Bronchoscopy in the diagnosis of bronchopulmonary cancer]. PMID- 3031807 TI - [Characteristics of a case of pulmonary paraganglioma (pheochromocytoma)]. PMID- 3031808 TI - [From the proceedings of the 13th National Conference of Pneumophtisiology. Bucharest, Oct. 1985]. PMID- 3031809 TI - Schistosoma mansoni: effects of anesthetics and antimonial drugs on worm shift in the mouse. PMID- 3031810 TI - [10 years of poliomyelitis in Spain (1976-1985)]. PMID- 3031811 TI - Characterization of leukotriene B4-omega-hydroxylase activity within human polymorphonuclear granulocytes. AB - Human polymorphonuclear granulocytes (PMN) metabolize exogenous [3H]leukotriene B4 (LTB4) into 20-hydroxy- and 20-carboxy-[3H]LTB4. The conversion was enhanced at acidic pH values (pH 6.0-7.0). Sonication of purified PMN and subcellular fractionation by differential centrifugation showed that major LTB4-hydroxylase activity was associated with the microsomal fraction (105,000 g pellet). In contrast to intact cells, LTB4-hydroxylase activity within the microsomal fraction revealed optimal activity at neutral pH and was inhibited by a wide range of divalent cations. There was a strict requirement for the presence of suitable electron donors such as NADPH. Heterocyclic nitrogenous bases, such as imidazole and pyridine, inhibited the LTB4 conversion induced by intact PMN as well as by their microsomes. These observations combined with the spectrophotometric analysis (carbon monoxide dithionite-reduced difference spectrum) supported the assumption that LTB4-hydroxylase resembled a cytochrome P 450 enzyme. The LTB4-hydroxylase within human PMN was not identical with the cytochrome P-450 of rat liver; hepatic microsomes only showed minute conversion of LTB4. PMID- 3031812 TI - In vitro activity of ampicillin alone and in combination with different concentrations of 6 beta-bromopenicillanic acid, clavulanic acid and mecillinam. AB - The antibacterial activity of ampicillin against Enterobacteriaceae strongly increased when combined with 6 beta-bromopenicillanic acid (BPA) or clavulanic acid (CA). In vitro, the combination ampicillin: BPA in the ratio 1:1 proved to be the most effective one. The antibacterial activity of mecillinam against Enterobacteriaceae was strongly potentiated by the addition of ampicillin. The combination mecillinam:ampicillin in the ratio 4:5 showed an antibacterial activity comparable to that of new beta-lactam compounds such as cefotaxime and ceftazidime. PMID- 3031813 TI - [Loeffler's endocarditis and disseminated viral infection]. AB - We report a case of Loffler's endocarditis in a 60-year-old woman with multiple lymphadenopathies, blood eosinophilia and acute heart failure. The eosinophilia developed over a six-month period to a maximum of 54.5% eosinophiles for 7600 X 10(9) leukocytes/l. Five weeks before death lymph node biopsy revealed Hodgkin's disease with mixed cellularity. Autopsy disclosed parietal endocarditis with endomyocardial fibrosis and mural thrombi, principally in the inflow tract of the left ventricle, lesions typical of the fibroplastic parietal endocarditis described by Loffler. Examination of the lymph nodes at autopsy showed a disseminated viral infection probably due to a virus of the herpes group, as was confirmed by electron microscopy. Since these findings disagree with the interpretation of the lymph node biopsy we think the diagnosis of Hodgkin's disease should be withdrawn. Loffler's endocarditis is discussed with reference to pathological features, differential diagnosis and lymph node changes in the patient. PMID- 3031814 TI - How photoreceptor cells respond to light. PMID- 3031815 TI - Survival from extreme lactic and keto-acidosis in diabetes mellitus. AB - A case report of extreme acidosis associated with diabetic metabolic decompensation is described. Treatment with conventional therapy and sodium bicarbonate resulted in complete recovery. PMID- 3031816 TI - Introduction of a normal human chromosome 11 into a Wilms' tumor cell line controls its tumorigenic expression. AB - The development of Wilms' tumor, a pediatric nephroblastoma, has been associated with a deletion in the p13 region of chromosome 11. The structure and function or functions of this deleted genetic material are unknown. The role of this deletion in the process of malignant transformation was investigated by introducing a normal human chromosome 11 into a Wilms' tumor cell line by means of the microcell transfer technique. These variant cells, derived by microcell hybridization, expressed similar transformed traits in culture as the parental cell line. Furthermore, expression of several proto-oncogenes by the parental cells was unaffected by the introduction of this chromosome. However, the ability of these cells to form tumors in nude mice was completely suppressed. Transfer of other chromosomes, namely X and 13, had no effect on the tumorigenicity of the Wilms' tumor cells. These studies provide support for the existence of genetic information on chromosome 11 which can control the malignant expression of Wilms' tumor cells. PMID- 3031817 TI - Visualization of viral clearance in the living animal. AB - The early events in viral dissemination via the bloodstream were identified by monitoring the fate of 123I-radiolabeled reovirus after it was injected intravenously in rats. Continuous scintillation camera imaging showed that reovirus serotypes 1 and 3 were cleared from the circulation in less than 10 minutes by specific and distinct target organs. Reovirus serotype 1 accumulated predominantly in the lungs and the liver, whereas serotype 3 accumulated in the liver and the spleen with very little virus uptake by the lungs. Incubation of reovirus serotype 1 with a monoclonal antibody directed against the viral hemagglutinin before injection totally inhibited the clearance of the virus by the lungs. Similar results were obtained when viruses biolabeled with 35S were used. These results demonstrate that viruses can be rapidly transported through the bloodstream to specific target organs and that the localization of the viruses depends on the interaction between specific viral surface components and the target organ. PMID- 3031818 TI - Platelet-mediated trigger mechanisms in the contact phase of blood coagulation. PMID- 3031819 TI - The role of arachidonic acid metabolites in preeclampsia. AB - Preeclampsia is characterized by imbalances in the production rates of several arachidonic acid metabolites. There is a considerable amount of evidence to indicate that PGI2 production is decreased in preeclampsia. PGI2 is a potent vasodilator, an inhibitor of platelet aggregation, and an inhibitor of uterine contractility. Evidence is accumulating that TX production in preeclampsia is either increased or unchanged so that the ratio of TX to PGI2 favors TX. TX opposes the actions of PGI2 in that it is a potent vasoconstrictor, a stimulator of platelet aggregation, and a stimulator of uterine contractility. Therefore, an imbalance of increased TX/decreased PGI2 production can account for the major clinical symptoms of preeclampsia. There is now preliminary evidence that the placenta produces lipoxygenase, as well as cyclooxygenase compounds. The hydroxyeicosatetranoic acids (5-HETE, 12-HETE, 15-HETE) and LTB4 have been identified as placental products. The preeclamptic placenta appears to be deficient in the 5- and 12-lipoxygenase enzymes, as indicated by placental production of 5-HETE and 12-HETE. Little is known about the regulation of arachidonic acid metabolites in normal or preeclamptic pregnancies. Steroids are known to affect cyclooxygenase product production. Progesterone, for example, inhibits PGI2 production. Although circulating and urinary concentrations of progesterone do not differ between normal and preeclamptic pregnancies, there is one report that placental progesterone concentrations are higher in preeclampsia; this may partially explain the decreased placental production of PGI2. There is a considerable amount of evidence in various tissues that regulatory interactions exist between the cyclooxygenase and lipoxygenase metabolites of arachidonic acid, but the role of these interactions in preeclampsia is not known. There is still much to be learned about the etiology of preeclampsia, but it is certain that aberrations in the production of arachidonic acid metabolites play an important role. It is likely that the effective treatment of preeclampsia will come from understanding the regulation of the arachidonic acid metabolites during pregnancy, and, therefore, the ability to specifically treat the production imbalances that characterize this disorder. PMID- 3031820 TI - Prostaglandin and leukotriene receptors in pulmonary, vascular, and uterine smooth muscle. AB - Prostaglandins are currently used to maintain patency of the ductus arteriosus and to elicit uterine contractions. Prostaglandin synthesis inhibitors are used to promote closure of the ductus and their administration in pregnant animals has produced fetal pulmonary hypertension. Exposure of the human fetus to inhibitors of prostaglandin synthesis has been associated with persistent pulmonary hypertension. A case report of a child with primary hypertension supports the hypothesis that the balance of prostaglandin metabolites plays an important role in maintaining PVR. Leukotrienes have been identified in the sputum of allergic asthmatic patients, patients with cystic fibrosis and infants with persistent pulmonary hypertension. Leukotriene inhibition in rats and newborn lambs prevented and reversed HPV. Receptors for prostaglandins have been identified in smooth muscle preparations of the uterus and renal glomerulus. Further studies have characterized the binding sites in lung tissue, giving supportive evidence for the existence of receptor sites there. Specific receptor sites for LTC4 and LTD4 have been demonstrated in lung tissue. Temperature, pH, and the presence of cations and guanine nucleotides have been shown to affect the receptor density and affinity. Lewis et al demonstrated that the characteristics of the receptor for [3H]LTD4 in the human lung are identical in adult and fetal tissue. This leads to the need for further investigation of the receptors and the effects of the local environments in an attempt to explain the physiologic changes seen in the successful and unsuccessful transition from fetal to neonatal circulation. PMID- 3031822 TI - [Pneumonectomy with atrial resection in patients with cancer of the lung]. PMID- 3031821 TI - Isolation and characterization of a cDNA for the catalytic subunit of the (Na+ + K+)-ATPase. PMID- 3031823 TI - [Effect of hemosorption on the status of neuromuscular transmission in myasthenia]. PMID- 3031824 TI - [Specific chemotherapy of herpetic genital lesions]. PMID- 3031825 TI - Effects of Bloom's syndrome fibroblasts on genetic recombination and mutagenesis of herpes simplex virus type 1. AB - The effects of Bloom's syndrome (BS) fibroblasts on genetic recombination and the mutation frequency of herpes simplex virus type 1 (HSV-1) was determined by employing two factor crosses of selected temperature-sensitive (ts) mutants. A significant increase in the recombination frequency (RF) was observed in seven of nine crosses when multiple BS fibroblast lines were compared to normal human fibroblasts. The RF of HSV-1 ts mutants increased following 12-O tetradecanoylphorbol-13-acetate (TPA) treatment of normal, but not BS fibroblasts, suggesting that BS fibroblasts express higher constitutive levels of genetic recombination activity. HSV-1 ts mutants demonstrated significantly higher reversion frequencies to the nontemperature sensitive (ts+) phenotype following growth in BS rather than normal fibroblasts, indicating that exogenous viral DNA encoding many of the enzymes necessary for its own replication is affected by the mutator phenotype of BS. PMID- 3031826 TI - Glucocorticoid-dependent maturation of viral proteins in mouse lymphoma cells: isolation of defective and hormone-independent cell variants. AB - Maturation of mouse mammary tumor virus proteins is dependent on glucocorticoid hormones in W7MG1, a stably infected mouse T-lymphosarcoma cell line derived from WEH17. We used an immunological procedure to select variant cell lines with altered levels of viral glycoproteins on the cell surface. One variant, W7M329a, expressed lower-than-normal levels of the major viral glycoprotein, gp52env, on the cell surface before and after exposure to hormone. Two other variants, W7M302b and W7M326.4, expressed elevated levels of gp52env on the cell surface even in the absence of hormone. Analysis of the levels and/or rates of synthesis of viral RNA and glycoproteins before and after hormone treatment indicated that the variant phenotypes resulted from changes in posttranslational steps of viral gene expression. The hormone-independent maturation of MMTV proteins is a novel variant phenotype that has not previously been reported. PMID- 3031827 TI - Parasexual analysis of human pepsinogen molecular heterogeneity. AB - Pepsinogens (PGA) are the inactive precursors of pepsin, the major acid protease found in the stomach. Highly polymorphic variation of these proteins has been demonstrated in several populations, and comparison of the DNA restriction fragment patterns obtained from informative pepsinogen phenotypes suggest that the polymorphism results from chromosomal haplotypes containing variable numbers of pepsinogen genes. In order to isolate the three most common PGA haplotypes (A, B, and C) and to unambiguously demonstrate their relationship to the observed protein heterogeneity, we constructed mouse X human somatic cell hybrids from individuals heterozygous for PGA and INS (insulin). Here, we describe analysis of hybrid cell lines that segregated human chromosomes containing the PGA genes and thereby provided for the parasexual discrimination of the different haplotypes on chromosome 11 determining the corresponding heterozygous phenotypes. These studies demonstrate that the A, B, and C haplotypes contain three, two, and one PGA genes, respectively. This unusual polymorphism of genomic DNA encoding very similar proteins probably reflects recent evolution by gene duplication. PMID- 3031828 TI - [Surgical treatment of small-cell forms of lung cancer]. PMID- 3031829 TI - Langerhans' cells, papillomaviruses and oesophageal carcinoma--a hypothesis. PMID- 3031830 TI - Oesophageal carcinoma--search for a possible viral aetiology using molecular hybridisation techniques. PMID- 3031831 TI - Malignant fibrous histiocytoma of the spermatic cord. A case report. AB - A case of malignant fibrous histiocytoma of the spermatic cord treated by primary local excision, followed 5 days later by radical inguinal orchidectomy and hemi scrotectomy is reported. At 30 months' follow-up the patient is free from local recurrence. PMID- 3031832 TI - Epstein-Barr virus reactivation in renal transplant recipients. AB - Renal transplant recipients at Tygerberg Hospital were investigated to determine the importance of Epstein-Barr virus (EBV) as a pathogen in these patients. All 106 patients investigated were shown to have EBV antibodies before transplantation and most had serological evidence of reactivation of the infection after transplantation. A mild clinical illness was present in a few patients concomitant with EBV reactivation, which may suggest that this virus has a role in the morbidity of some renal transplant recipients. Lymphoblastoid cell lines were established from 11 renal transplant recipients; 5 of these cell lines were shown to be virus producers and 1 is thought to have unique properties. PMID- 3031833 TI - Herpes simplex virus shedding in genital secretions. AB - Nine women and eight men with a history of genital herpes had cultures taken for 60 consecutive days to assess the frequency and duration of viral shedding in the absence of symptoms. Daily self-obtained specimens (cervical-vaginal swabs from women and urethral swabs from men) were submitted for viral isolation. Five of 972 samples were found to contain infectious virus; two of the positive specimens correlated with overt disease. The other three positive cultures, two from the same individual, were not clearly associated with a genital infection. Thus, the overall frequency of asymptomatic viral shedding was 0.31% and occurred in two of 17 individuals. It is concluded that prolonged continuous sampling may be necessary to assess the risk of asymptomatic shedding. Infectious virus was not present in the genital secretions of most individuals (15 of 17) in the absence of a lesion, even when cultured on a daily basis for 60 days. PMID- 3031834 TI - Acquisition of concomitant oral and genital infection with herpes simplex virus type 2. AB - Primary oral and genital infections caused by herpes simplex virus type 2 were diagnosed in an 18-year-old female. A history of sexual practices was critical in determining the anatomic sites of infection. Restriction endonuclease analysis of viral DNAs helped to identify the male sexual partner from whom the virus had been acquired. He had been infected recently by a previous sexual partner but had not yet developed lesions. Clinicians should obtain a history of sexual practices from patients with newly acquired genital herpes and should advise patients with genital herpes that transmission of virus to sexual partners can occur in the absence of overt lesions. PMID- 3031836 TI - Update on the diagnosis and treatment of rare neuroendocrine tumors. AB - A growing knowledge and awareness of bizarre clinical presentations, the physiology of new peptides, and the more frequent use of radioimmunoassay techniques has led to the identification of more patients with glucagonomas, somatostatinomas, vipomas, and ectopic tumors. Definite clinical syndromes, though sometimes seemingly "silent," occur as a result of hypersecretion of newly identified hormones and neurotransmitters that act in endocrine, neurocrine, or paracrine fashion to alter normal metabolism of carbohydrates and electrolytes. Metabolically and clinically, glucagonomas are catabolic, somatostatinomas are inhibitory, and vipomas are diarrheogenic. The clinical syndromes can be differentiated from other, more common, endocrinopathies; the measurement of the plasma concentrations of the specific peptides is not only diagnostic, but prognostic. Specific pathologic confirmation of the functional potential of these tumors by immunocytochemical techniques is now possible. The goal of diagnosis is detection and tumor localization before metastases occur so that surgical excision may be curative. Objective clinical and humoral responses to chemotherapy for nonresectable or metastatic lesions can be expected in about 50 per cent of patients; specifically, dacarbazine (DTIC) is the agent of choice for glucagonomas, streptozotocin for somatostatinomas, and leukocyte interferon for vipomas. PMID- 3031835 TI - Factors influencing the growth of normal and neoplastic thyroid tissue. AB - TSH activates the adenylate cyclase and the phosphoinositide turnover-protein kinase C-calcium systems in thyroid cells and appears to have an important but variable role in controlling the growth of both normal and neoplastic thyroid tissues. Species differences, experimental conditions, and tissue or tumor cell heterogeneity may account for this variability. Although TSH seems to be an important physiologic growth factor, it is neither the exclusive growth factor for the thyroid gland nor absolutely necessary for the effect of other thyroid growth factors. TSH may work in concert with other growth factors such as EGF. Some growth factors influence thyroid growth through TSH, whereas others do not. PMID- 3031837 TI - Potent inhibition of superoxide anion generation by PGE1 and the PGE1 analogue OP 1206 in human PMN's--unrelated to its antiplatelet PGI2-like activity. PMID- 3031838 TI - [Hyperfractionated irradiation of bronchial cancer. Results of a pilot study]. AB - The results achieved during recent years in experimental radiotherapy of malignant tumors show more and more the benefit provided by the small individual doses applied in fractionated irradiation. The effects and side effects of a hyperfractionated therapy of the bronchial carcinoma were investigated in a not randomized comparative study. The data of 100 patients were available for evaluation; they showed a tendency to local superiority of hyperfractionated irradiation. The value of hyperfractionation shall be examined in detail in a randomized prospective study. PMID- 3031839 TI - [Results of irradiation using different radiation sources in the treatment of malignant parotid tumors]. AB - Patients with malignant parotid tumors were treated at two different centres with slightly ionizing radiation (110 patients) and neutron therapy (15 patients). The treatment results are compared. The principles of comparison applied in this analysis are identical. The histologic classification was made according to the W.H.O. recommendation of 1972 and the staging according to Becske. Local tumor control is the criterion applied in the comparative assessment of both methods. The analysis has shown that there is no significant difference in the treatment results within the individual stages of tumor's advancement after application of an orthovolt therapy and a telecobalt therapy. Patients treated in early stages (stages I and III) had a substantially higher rate of local tumor control (72%) than patients in advanced stages (46%, stages III and IV). A more favorable treatment results was achieved by neutron therapy in all stages of tumor's advancement. In this case the rate of patients with local tumor control (87%) was significantly higher than after radiotherapy with slightly ionizing radiation (46%). PMID- 3031840 TI - Misonidazole combined with radiotherapy in the treatment of non-small cell lung cancer. A randomized double-blind trial. AB - Forty-one patients with inoperable non-small cell lung cancer were randomized to receive irradiation therapy 45 Gy in ten fractions over eight weeks, plus either placebo or misonidazole 1.2 g/m2 orally on treatment day. The irradiated area was the primary tumour. Twenty-one patients received misonidazole and 20 received placebo. Minimal observation time for the study was 36 months. No clear difference was observed in the pattern of relapse between the groups. 43% of the patients in the misonidazole group attained a complete response (CR) or partial response (PR) as compared with 65% in the placebo group. Median survival time was twelve months in the misonidazole group and 14 months in the placebo group. PMID- 3031841 TI - Interleukin 1 stimulates endothelial cell tissue factor production and expression by a prostaglandin-independent mechanism. AB - Activation of coagulation occurs at inflammatory sites following the ingress of mononuclear cells, and may result from alterations in the vessel wall. Since the monokine, interleukin 1, initiates diverse responses to inflammation, its ability to enhance vascular procoagulant activity was studied. Interleukin 1-treated cultured human endothelial cells acquired elevated levels of the procoagulant, tissue factor. This required de novo protein synthesis, was maximal at 2 h after exposure to interleukin 1, and resulted in persistently elevated cellular procoagulant activity. Tissue factor was later expressed (6-24 h) on the surface of uninjured endothelial cells. Endothelial cell procoagulant production and expression in response to interleukin 1 could be dissociated from endogenous prostaglandin metabolism, being insensitive to hydrocortisone, indomethacin, eicosatetrayionic acid and exogenous arachidonic acid. In addition, no increase in prostaglandin synthesis occurred during the interval in which tissue factor was synthesized. We therefore conclude that interleukin 1 stimulates endothelial synthesis and surface expression of tissue factor by a prostaglandin-independent mechanism. PMID- 3031842 TI - [2 newborn infants with severe arrhythmia caused by hyperkalemia]. AB - Two newborn infants with ventricular arrhythmias secondary to hyperkalaemia are presented. One child also showed a decreased serum calcium concentration. There is scanty literature concerning the often life threatening cardiac arrhythmias due to hyperkalaemia in the newborn infants. Treatment of the cardiac arrhythmias require intravenous calcium gluconat and sodium bicarbonate infusion beside lowering the serum potassium level in the usual way. PMID- 3031843 TI - Effect of microinjected amine and diamine oxidases on the ultrastructure of eukaryotic cultured cells. AB - Diamine oxide and serum amine oxidase, which catalyse the oxidation of diamines and polyamines, respectively, were trapped within reconstituted Sendai virus envelopes. These loaded envelopes were incubated with cultured normal chick fibroblasts or with fibroblasts transformed by Rous sarcoma viruses. The binding of the reconstituted envelopes to the cultured cells was confirmed by scanning electron microscopy. It has been shown that the reconstituted envelopes (1-3 microns diameter) were attached to the eukaryotic cells. No significant changes in the morphology of the normal chick embryo fibroblasts were noted upon treatment with enzyme-loaded envelopes. On the other hand, chick embryo fibroblasts transformed by Rous sarcoma virus were affected by the microinjected amine oxidases. Scanning electron microscopy demonstrated the formation of holes in the microinjected cells. Similar morphological changes were also observed when diamine oxidase was microinjected into cultured glioma cells. These holes may be the result of the ejection of the nucleus. These findings are in line with the observed effect of the injected amine oxidases on macromolecular synthesis in normal and transformed chick embryo fibroblasts. PMID- 3031844 TI - [After care: who needs it?]. PMID- 3031845 TI - Tumor histology and the immunoregulatory T lymphocyte subsets in lung cancer patients. AB - The immunoregulatory T lymphocyte subsets (CD4+, CD8+) in 39 patients with primary lung cancer were analyzed as for the possible correlation with tumor histology. Compared to the healthy controls (n = 36), cancer lymphocytes were generally characterized by a higher proportion of CD8+ subset (p less than 0.01) with no difference in either CD4+ subset or CD3+ cells. There was, however, a significant difference (p less than 0.05) between one extreme of epidermoid carcinoma with the highest CD8+ and the lowest ratio (CD4+/8+) and the other end of adenocarcinoma with the lowest CD8+ and the highest ratio. Large cell and small cell carcinomas were of the intermediate values. A spectrum of the subset marker profiles was thus found in associated with tumor histology. A diversity in the host immune status was suggested in primary lung cancer, although clinical implication remains to be clarified. PMID- 3031846 TI - Studies on the mechanism of carbon monoxide-induced vasodilation in the isolated perfused rat heart. AB - We investigated the effects of dissolved CO on isolated potassium-arrested (K+) perfused rat hearts. Hearts from male Sprague-Dawley rats were perfused via the aorta with oxygenated Krebs-Henseleit solution containing 20 mM K+. Coronary flow (Qt) averaged 48.8 +/- 1.6 (SE), 48.1 +/- 1.7, and 55.6 +/- 1.7 ml/min/g dry wt when the perfusate was equilibrated with 95% O2-5% CO2, 5% N2-90% O2-5% CO2, and 5% CO-90% O2-5% CO2, respectively. The change in Qt was statistically significant when CO was present in the perfusion medium, but was not significant when N2 was present. Furthermore, the effect was reversible because coronary flow returned to control levels when CO was removed. Myocardial oxygen consumption (MVO2) did not change significantly when hearts were perfused with either N2 or CO. The magnitude of CO-induced vasodilation was not affected significantly by the addition of either 5 microM propranolol, 2 microM phentolamine, 1 unit of adenosine deaminase, or 0.1 mM indomethacin to the perfusate. In addition, CO reversed the vasoconstrictive effects of the alpha-agonist methoxamine. These results indicate that CO exerts a vasodilatory effect on coronary vasculature that is not the result of decreased O2 content in the perfusate and is not mediated by adrenergic influences, adenosine, or prostaglandins. PMID- 3031847 TI - Primary afferent terminal function following acrylamide: alterations in the dorsal root potential and reflex. AB - The dorsal root potential (DRP) and the dorsal root reflex (DRR) were studied in acrylamide (ACR)-induced axonopathy to determine the nature and extent of primary afferent terminal (PAT) dysfunction. Cats were administered 30 mg/kg/day ACR for either 5 (ACR 5D) or 10 (ACR 10D) days. The day after the last injection, the spinal cord as isolated in situ and the DRP and DRR were elicited. It was found that only 21% of the ACR 10D animals exhibited a DRP. Furthermore, in that 21%, the DRP appeared to degrade over distance differently from the control group. There was no change in the DRP evoked in the ACR 5D group. ACR affected the DRR when evoked from the cutaneous sural nerve (SU) to a greater extent than when evoked from the medial gastrocnemius (MG) nerve. A SU-evoked DRR could not be elicited in any of the animals in the ACR 10D group and in only 20% of the ACR 5D group. There was no difference in the ability to elicit a MG-evoked DDR in either ACR-treated group when compared to control. However, the maximum-evoked area under the DRR elicited from the MG nerve was significantly smaller in the ACR treated groups than in control. These data show that ACR does indeed impair PAT function. ACR preferentially affects the PAT processing of the SU when compared to the MG nerve, which may indicate that the selective vulnerability of the largest diameter fibers to ACR is not necessarily true. PMID- 3031848 TI - The copper-induced deformability loss and echinocyte formation in human erythrocytes: an electron paramagnetic resonance study. AB - The effects of incubating human erythrocytes with CuSO4 under prelytic conditions were investigated to clarify the cause of the decreased whole-cell deformability in flow, using electron paramagnetic resonance and the spin label methods. The effect of Cu(II) on erythrocytes was manifested by the dose-dependent echinocyte formation as well as by the decreased whole-cell deformability in flow. The deformability loss was attributed in part to the altered rheological characteristics due to echinocytosis. By using various reagents capable of reversing the effects of Cu(II) on morphology and deformability loss, it was shown that at least two types of damage sites can be distinguished. They include (1) the Cu(II) coordination sites which can be dissociated by chelating reagents and (2) the sites involving sulfhydryl groups in which disulfide bonds and probably Cu(II)-bridged strong coordination bonds are formed. The latter can be reversed by sulfhydryl reagents such as dithioerythritol, but not by EDTA or penicillamine. Pretreatment of erythrocytes with iodoacetate eliminates the variance in susceptibility to Cu(II) among donors and generally enhances the effects of Cu(II) by lowering the cellular glutathione level. A possible relation between the nature of the damage sites and the cellular glutathione level is discussed. PMID- 3031850 TI - Lead-induced inclusion bodies in rat kidney after perinatal treatment with lead and disulfiram. AB - The presence of inclusion bodies in renal proximal tubules was studied in rats exposed to lead and/or disulfiram (tetraethylthiuram disulfide). Pregnant rats were treated with only lead acetate (0.25% Pb in the drinking water), only disulfiram (0.1 mmol/kg p.o. twice a week) or with both lead acetate and disulfiram from day 1 of pregnancy and until the offspring were 4 weeks of age. After parturition the disulfiram was given s.c. directly to the offspring instead of to the dams. Treatment was discontinued at weaning and tissue samples from renal cortex were studied by electron microscopy. In lead-treated dams inclusions were present in nuclei of renal proximal tubule cells in the 3 segments with the highest incidence in the middle segment. Inclusions were also present in the cytoplasm. In the offspring, indirectly exposed to lead via the dams, inclusions were present in all 3 segments. No inclusions were present in control rats or in disulfiram-treated rats. Combined treatment with lead and disulfiram resulted in a marked decrease in the incidence of inclusion bodies both in the dams and in the offspring compared to in rats treated with only lead. Diethyldithiocarbamate, a major metabolite of disulfiram, forms a lipophilic complex with lead, and is known to cause pronounced effects on the tissue distribution of lead. The present investigation shows that lead inclusion bodies are formed in the offspring indirectly exposed to lead via the dams during gestation and lactation. Concurrent exposure to disulfiram reduces the incidence of inclusion bodies in renal proximal tubules, probably due to formation of a lead-dithiocarbamate complex. PMID- 3031849 TI - Amantadine-induced lipidosis. A cytological and physicochemical study. AB - The purpose of this study was to test whether or not the antiviral drug amantadine induces the structural features of lipidosis in intact animals (rats) and cultured cells, and to investigate the interactions between amantadine and phospholipids. Chlorphentermine was used as reference compound. When subchronically fed to rats at a daily dosage of approximately 180 mg/kg, amantadine induced ultrastructural symptoms of generalized lipidosis, the degree of which was, however, by far less marked than that previously reported for chlorphentermine. This was paralleled by the findings on cell cultures (rat peritoneal macrophages), where the lipidosis-inducing potency of amantadine was approximately 10-fold lower than that of chlorphentermine. As to drug phospholipid interactions, amantadine had less marked effects than chlorphentermine upon the phase transition characteristics of phosphatidylcholine and phosphatidic acid; furthermore, amantadine was approximately 10-fold less potent than chlorphentermine in displacing Ca from phosphatidylserine monolayers. The present study has revealed a parallel between the comparatively low lipidosis inducing efficacy inherent to amantadine and the comparatively low tendency to interact with phospholipids. It is suggested that the cage-like structure of the amantadine molecule hinders an effective intercalation of the drug into phospholipid aggregates, and that this is an essential factor responsible for the low inherent efficacy of amantadine with respect to lipidosis induction. PMID- 3031851 TI - Synergistic interactions of 2,3,7,8-TCDD and 2,2',4,4',5,5'-hexachlorobiphenyl in C57BL/6J and DBA/2J mice: role of the Ah receptor. AB - Treatment of C57BL/6J mice with 2,2',4,4',5,5'-hexachlorobiphenyl (HCBP, 500 mumol/kg) elevated hepatic cytosolic Ah receptor levels 82-107% for up to 14 days. Scatchard analysis of the [3H]2,3,7,8-TCDD (TCDD)-Ah receptor saturation binding curves from corn oil and HCBP treated rats gave KD values of 0.80 and 0.90 nM, respectively and confirmed that treatment with HCBP did not significantly alter receptor-radioligand affinities. Administration of HCBP to DBA/2J mice did not result in detectable hepatic cytosolic Ah receptor levels. Cotreatment of C57BL/6J mice with HCBP (500 mumol/kg) at a dose level of TCDD (1 nmol/kg) which elicited less than 10% of the maximum induction response resulted in significant synergistic induction of hepatic EROD and AHH [compared to animals treated only with TCDD (1 nmol/kg)]. In contrast, cotreatment of C57BL/6J mice with HCBP (500 mumol/kg) and maximally inducing dose levels of TCDD (100 or 500 nmol/kg) resulted in either a slight or no difference in the induction of AHH or EROD compared to the induction responses observed in mice treated only with TCDD. In contrast, cotreatment of DBA/2J mice with TCDD and HCBP (500 mumol/kg) resulted in significant synergistic induction of AHH and EROD at both submaximal (10-500 nmol/kg) and maximal (5000 nmol/kg) induction levels of TCDD. The only significant interactive effect of HCBP (500 mumol/kg) on the toxicity of TCDD in C57BL/6J and DBA/2J was protection from body weight loss observed after cotreatment of HCBP and TCDD in DBA/2J mice. PMID- 3031852 TI - Platelet aggregation inhibitors from Agkistrodon acutus snake venom. AB - Among all the purified components from A. acutus venom, including ADPase, 5' nucleotidase, phospholipase A2 and fibrinogenases, only the venom ADPase (50-100 micrograms/ml) shows marked inhibitory action on ADP (10 microM)-, collagen (10 micrograms/ml)- and sodium arachidonate (100 microM)-induced platelet aggregations of rabbit platelet-rich plasma. The venom 5'-nucleotidase (100 micrograms/ml) inhibited ADP-induced platelet aggregation by 31 +/- 4% (n = 4, P less than 0.05). Fibrinogenolytic enzymes (fractions I and IX, 100 micrograms/ml) did not significantly inhibit platelet aggregation induced by ADP (10 microM), collagen (10 micrograms/ml) or sodium arachidonate (100 microM). However, when the fibrinogenase (fraction IX, 100 micrograms/ml) was preincubated with platelet rich plasma for 30 min it inhibited collagen (20 micrograms/ml)- and ADP (10 microM)-induced platelet aggregations by 34 +/- 9% (n = 4, P less than 0.05) and 35 +/- 6% (n = 4, P less than 0.05), respectively. The phospholipase A2 (100 micrograms/ml) did not affect platelet aggregation. The venom ADPase is a single chain polypeptide with a molecular weight of 94,000. The specific ADPase activity is estimated to be 4.3 mu moles Pi/min/mg of protein. It also possesses phosphodiesterase and weak 5'-nucleotidase activities. PMID- 3031854 TI - Isopropyl methanesulfonate is a potent red cell hematotoxicant. AB - Mice given 10-weekly injections (i.m.) of 20 mumol of isopropyl methanesulfonate (IPMS) in trioctanoin exhibited depressed hematopoiesis as manifested by reduced peripheral blood counts and by reduced thymic and bone marrow cellularities. The treatments were particularly toxic to erythroid populations and ultimately induced signs of splenic erythropoiesis. Bone marrow cellularity recovered during the latter stages of the treatments indicating that subpopulations of hematopoietic cells were resistant to the treatments. This recovery of bone marrow cellularity may bear on the recently discovered leukemogenic potential of IPMS. PMID- 3031853 TI - Chromosome aberrations and sister-chromatid exchanges induced by technical grade toluene diisocyanate and methylenediphenyl diisocyanate in cultured human lymphocytes. AB - Technical-grade toluene diisocyanate (TDI; 80% 2,4-isomer and 20% 2,6-isomer) and 4,4'-methyl-enediphenyl diisocyanate (MDI; about 45% MDI, 25% 4,4' methylenediphenyl triisocyanate and 30% unspecified compounds of higher molecular weight), used as hardeners in the production of polyurethane, induced chromosome aberrations after a 24 h treatment in the absence of metabolic activation in human whole-blood lymphocyte cultures, MDI at all doses tested (0.54-4.30 microliter/ml) and TDI (0.019-0.150 microliter/ml) at the highest two doses (0.075 and 0.150 microliter/ml). In the presence of rat liver S9 mix (1.5 h treatment), both mixtures significantly increased aberrations at only one of the doses used, MDI at the highest dose (4.30 microliter/ml) and TDI at the second highest dose (0.038 microliter/ml). MDI also marginally increased sister chromatid exchanges at the highest dose available (2.17 microliter/ml) with and without (48 h treatment) S9 mix. On addition to culture medium, both TDI and MDI formed polymers, which were seen as small particles on the microscopic slides. At high doses the presence of these polymers made metaphase analysis impossible. PMID- 3031855 TI - Two dimensional gel analysis of HLA-DR2 associated HLA-D clusters. AB - Five cell lines with different cellular HLA-D specificities associated with HLA DR2, i.e. Dw2, Dw12, MN2, DB9 and D2J, displayed structural variation in the products of the DR and DQ loci. Analyses of immunoprecipitated DR molecules from HLA-D homozygous B-lymphoblastoid cell lines by two dimensional polyacrylamide gel electrophoresis showed variable patterns with one set of DR beta chains. Another set of DR beta chain products, however, was homogeneous in all cell lines studied. In addition, analyses of DQ molecules showed extremely variant DQ beta chains, and slightly variant DQ alpha chains. PMID- 3031856 TI - [Chemodectomas of the neck]. PMID- 3031857 TI - Radioimmunoassay of urinary 3 beta-hydroxy-5-cholenoyl glycine in hepatobiliary disease. AB - A radioimmunoassay for 3 beta-hydroxy-5-cholenoyl glycine in human urine has been developed. The antiserum was elicited with the antigen in which the steroid hapten is linked to a bovine serum albumin through the C-19 position. The [125I] tyrosine derivative of the hapten was used as radioligand. The standard curves were linear ranging from 10 to 320 ng/mL. The cross-reactivities with other bile acids were not detectable and below 0.3% with cholesterol. Sample preparation includes extraction of 3 beta-hydroxy-5-cholenoyl glycine from urine and solvolysis of the sulfates--main form present in urine. Urinary excretion of 3 beta-hydroxy-5-cholenoyl glycine was 0.373 +/- 0.133 mumol/day in healthy adults. Urinary excretion of 3 beta-hydroxy-5-cholenoyl glycine increased in chronic liver dysfunction, hepatoma and obstructive jaundice in this order. PMID- 3031858 TI - Development of malignancies in Japanese renal transplant recipients. PMID- 3031859 TI - Primary hepatocellular carcinoma in Africans. PMID- 3031860 TI - [Correlations of the individual levels of the activity and heat resistance of energy-support enzymes in the lake frog]. AB - A study was made of the correlation between the levels of the activity and heat resistance of Na,K-ATPase, Mg-ATPase and succinate dehydrogenase (SDG) in Rana ridibunda. A correlation was observed between the activities of two ATPases, and between either of them and the activity of liver SDG. The correlation coefficient between the heat resistance levels was statistically significant only in SDG of liver and muscles. The question whether the criterion of T50% inactivation may be good for a comparative appraisal of the heat resistance of enzymes is under discussion. PMID- 3031861 TI - Cystosarcoma phyllodes. A malignant cystosarcoma phyllodes in a 14-year-old girl. PMID- 3031862 TI - Neoplastic potential in patients with disorders of sexual differentiation. AB - The occurrence of tumors was determined in 157 patients with disorders of sexual differentiation at a major pediatric hospital from 1960 to 1980. Gonadoblastoma developed in 2 of the 4 patients with mixed gonadal dysgenesis (MGD) who had not had bilateral gonadectomy; this indicates the need for early gonadectomy in such cases. Gonadal embryonal carcinoma developed in 1 child with Turner's syndrome (1 of 71). Four cases of nephroblastoma occurred, 1 in a child with MGD (1 of 18) and 3 in male pseudohermaphrodites (3 of 35). The nature of the defect linking male pseudohermaphroditism with Wilms tumor remains to be established. PMID- 3031863 TI - [Cyclic nucleotides in anterior chamber humor in primary glaucoma]. PMID- 3031864 TI - [Clinico-topographo-anatomical classification of juvenile angiofibromas of the nasopharynx]. PMID- 3031866 TI - [Comparison of the biological properties of various strains of Aujeszky's disease viruses]. AB - We compared three strains of Aujeszky's disease virus that had been isolated from slaughter pigs from the dormant foci of Aujeszky's disease, with the known virulent strain CVOS and TOP. No significant differences were found out in the following characters: titer value on tissue cultures from chick embryonal cells (KEB) and cellular line MDBK at the temperatures of 37 degrees C and 40 degrees C, cytopathogenic effect (CPE) in the HeLa cells, plaque formation in the KEB tissue culture, pruritus rise and the time of rabbit death after infection. Assessing the results of the comparison, the three new-isolated strains of Aujeszky's disease virus can be evaluated as strongly virulent. PMID- 3031865 TI - The zoonotic potential of feline leukemia virus. AB - The many studies that have addressed the possibility for FeLV infection in human beings are reviewed in this article. Because of the many similarities between FeLV-induced immune system impairment in cats and retrovirus related acquired immune deficiency syndrome in man, these two conditions are discussed as well. PMID- 3031868 TI - The prevalence of antibodies of Brucella abortus, Dermatophilus congolensis and bovine leukaemia virus in Nigerian slaughter cattle. AB - In a pilot survey to compare the relative prevalence of three diseases in apparently healthy White Fulani Zebu (WFZ) cattle slaughtered in Nigeria, sera from 80 randomly selected animals with no significant gross lesions on ante mortem and post mortem inspection were examined for antibodies to Brucella abortus, Dermatophilus congolensis and bovine leukaemia virus. Of the samples screened, 5.0, 8.8 and 2.0% showed serological evidence for brucellosis, cutaneous streptothricosis and bovine leukosis respectively. PMID- 3031867 TI - Comparison of the effect of live Newcastle disease vaccine Clone 30 in broilers administered at day 1 or at day 7 and the effect of H120 vaccination at 17 days of age: a field experiment. AB - To analyse the results of a vaccination on the first day of age against Newcastle disease (ND) and on the 17th day of age against Infectious Bronchitis (IB) resp. with spray vaccines with Clone 30 and H120 vaccine. These vaccinations are compared in field circumstances with other vaccination methods. A serological examination and challenge test were used to be informed about the response and protection. From the present study the following conclusions can be drawn: Clear indications are obtained that following a spray vaccination against ND with Clone 30 vaccine of one-day-old broilers which possessed maternal antibodies, birds received a moderately good protection against ND, in spite of very low levels of HI antibodies. A spray vaccination against IB with H120 vaccine of broilers at 17 days of age gave some protection from two weeks after vaccination, however making a good conclusion about the protection is impossible and further investigation is required. PMID- 3031869 TI - Vaccination of swine against H3N2-influenza field isolates using the human Philippines-strain. AB - Intratracheal inoculation of 2 Belgian H3N2-influenza viral strains, isolated from sick swine in the field, caused high fever, anorexia and dyspnoea in unvaccinated swine. The strains are related to the human A/Port Chalmers/1/73 (H3N2)-strain. In a limited study, 2 subunit vaccines, both derived from the human A/Philippines/2/82 (H3N2)-strain, were tested for efficacy in protecting swine against these Belgian field isolates. Vaccine A was a commercial vaccine, vaccine B an experimental vaccine. For evaluation of the efficacy of the vaccines, clinical as well as virological parameters were used. It was found that 2 spaced injections of the experimental vaccine (B) resulted in very high serum hemagglutination-inhibition (HI) titres against the Philippines-strain. Nevertheless, only partial protection was obtained, as indicated by the milder clinical signs and the decreased viral replication at challenge. One injection of the experimental vaccine (B) and 2 spaced injections of the commercial vaccine (A) did not result in any protection at challenge, even though moderate HI titres against the Philippines-strain were obtained. It was concluded that if an H3N2 strain is included in vaccines for use in swine, a strain should be selected which is identical or very closely related to the strain(s) prevalent in the swine population of the country in which the vaccine will be used. PMID- 3031870 TI - Parasites as stressors: plasma cortisol responses of goats infected with the stomach worm Haemonchus contortus to exogenous corticotropin (ACTH). AB - Ten male, juvenile pigmy goats of similar age and weight were allocated randomly to two groups. Goats in one group were each inoculated with 20,000 infective larvae of Haemonchus contortus. The other group served as uninfected controls. Goats were housed together, and precautions were taken to avoid the creation of differential, between group, stressogenic circumstances. Body weights, nematode egg production, hematocrits, and clinical signs were monitored over a 61-day period following inoculation of larvae. On Days 59 and 61, adrenal response tests (ART) were conducted by measuring the levels of plasma cortisol before and 2 h after administration of porcine ACTH at the rate of 0.35 I.U. kg-1 body weight on Day 59 and 2.2 I.U. kg-1 on Day 61. Although the infections did not reduce body weights, they were 'heavy' on the basis of egg production, and led to significant reductions in packed erythrocyte volumes. There was no significant difference between the groups of goats in the responses to ART, indicating that the infections did not produce sufficient stress to reduce the ability of the adrenal cortex to respond to exogenous ACTH. PMID- 3031873 TI - Highly sensitive antigen detection procedures for the diagnosis of infectious bovine rhinotracheitis: amplified ELISA and reverse passive haemagglutination. AB - The sensitivity of an enzyme-linked immunosorbent assay (ELISA) for the detection of bovid herpesvirus 1 antigen was increased by up to 50-fold using the biotin avidin interaction to amplify the reaction, when compared with a simple sandwich ELISA. An alternative immunoassay, reverse passive haemagglutination (RPHA), had a similar sensitivity to the amplified ELISA, and was technically simpler to perform. Both the amplified ELISA and the RPHA could detect viral antigen in the nasal secretions of calves undergoing experimental primary infection with the virus from Day 3 to Day 7 after inoculation. Neither assay was as sensitive as virus isolation in cell culture and they failed to detect antigen in virus positive samples from the calves from 8 days after inoculation, and from vaccinated calves undergoing challenge infection. PMID- 3031871 TI - Hemagglutination inhibition and virus neutralizing response of rabbits inoculated with bovine herpesvirus 1 subunit vaccine. AB - The immunogenicity of bovine herpesvirus type 1 (BHV-1) hemagglutinin has been investigated. Both live and nonionic detergent solubilized vaccines were prepared and 5000 hemagglutinating units (HAU) were injected subcutaneously into rabbits. Both types of vaccine induced a good antibody response but live virus was four times more efficient in inducing hemagglutination inhibiting and neutralizing antibodies than either Triton X-100- or octylglucoside-solubilized subunit vaccine. Blotting analysis revealed that five proteins, of 105,000, 90,000, 74,000, 64,000 and 54,000 mol. wt, were recognized by the serum of vaccinated animals. Triton X-100-solubilized vaccine did not induce antibodies against the 105,000 and 64,000 mol. wt proteins, indicating the important role of VP 90,000 and VP 74,000 in hemagglutination and neutralization. The order in which antibodies to the different viral proteins were induced was VP 90,000, (VP 105,000, VP 64,000, VP 54,000) and VP 74,000. Our data indicate that VP 90,000 is the hemagglutinin. Using convalescent serum from intranasally infected animals, we could identify nine structural proteins for BHV-1; VP 105,000, VP 90,000, VP 74,000, VP 64,000, VP 54,000, VP 50,000, VP 47,000, VP 40,000 and VP 31,000. PMID- 3031872 TI - Four-layer enzyme immunoassay (EIA) detection of differences in IgG, IgM and IgA antibody response to Aujeszky's disease virus in infected and vaccinated pigs. AB - The use of the four-layer enzyme immunoassay (EIA) for the detection of IgG, IgM and IgA antibodies against Aujeszky's disease virus in blood and oropharyngeal swabs of infected and vaccinated pigs is described. Mean antibody titres obtained using the four-layer EIA were 6.1 and 3829 times higher compared with the indirect enzyme-linked immunosorbent assay (ELISA) and virus neutralization (VN) test, respectively. The VN test detected mainly IgG antibodies, while the IgM antibodies did not react. Using the EIA, the first antiviral antibodies in sera were demonstrated on Days 5-7 after infection or vaccination. Up to the 7th day, demonstrable antibodies were almost exclusively of the IgM class. In infected pigs high titres of IgM antibodies were still detected on Day 18, while in vaccinated animals they were absent by this time. Antibodies of the IgG class appeared in infected pigs sooner (Day 7) than in vaccinated pigs (Day 10) and reached higher mean titres. Antibodies of the IgA class were demonstrable from Day 10 only in samples from infected pigs. Similar antibody dynamics and distribution were detected in oropharyngeal swabs, except that the IgG and IgM titres were roughly 100 times lower than in sera. However, titres of IgA antibodies in oropharyngeal swabs were two times higher than in sera. The greatest differences between both groups of animals were recorded on Day 18; in the infected pigs, IgG, IgM and IgA antibodies were present in sera and oropharyngeal swabs at that time, while in vaccinated pigs only IgG antibodies were demonstrable. The effect of infection and vaccination on the pattern of the immune response as well as the importance of the detection of individual immunoglobulin classes for the specificity of the enzyme immunoassay are discussed. PMID- 3031874 TI - Detection of bovine virus diarrhea virus in cell culture using an immunoperoxidase technique. AB - An indirect immunoperoxidase staining technique was developed for identifying cell cultures infected with bovine virus diarrhea virus. Infected cell monolayers stained intensely while uninfected monolayers remained colorless. Immunoperoxidase staining was as sensitive as direct immunofluorescence in detecting endpoint dilutions of virus suspensions. Using the immunoperoxidase technique, infected monolayers were detectable by macroscopic, as well as microscopic, observation. PMID- 3031876 TI - [Specific protective immune mechanism in herpes simplex virus infection]. PMID- 3031877 TI - [Antibodies to enteric adenovirus (species 40 and 41) in the sera of children, young adults and the elderly]. PMID- 3031875 TI - An analysis of histology and DNA-ploidy in primary wilms tumors and their metastases and a study of the morphological effects of therapy. AB - In children with Wilms' tumours the length of survival is greatly influenced by success in preventing or controlling metastatic disease. The current study focuses on the morphological aspects of metastases when compared with the primary tumour. In 8 patients it appeared that blastema is the most likely component to metastasize, whereas epithelial and stromal components were hardly, if at all, represented in metastases. Furthermore, flow cytometric DNA ploidy determinations on 4 cases showed that both the primary tumours and the metastases had stemlines in the diploid and low aneuploid (hyperdiploid) range. Finally, in four cases the influence of therapy on morphology of the primary tumours was analyzed. In these cases blastema seemed to be the component most sensitive to therapy. Thus, blastema seems to play a central role in prognosis of Wilms' tumours; either reacting to therapy or, if insensitive, by metastasizing. PMID- 3031878 TI - [Isolation of enteric adenoviruses in cell cultures]. PMID- 3031879 TI - [Bran in the treatment of irritable bowel syndrome]. AB - The paper deals with the problem of using wheat bran in the treatment of patients with irritable colon syndrome expressed as spastic constipation. The effectiveness of the treatment with the wheat bran only (in a dose of 30-35 g/day), and in combination with drugs was comparatively studied in 105 patients. The bran fractions differing in the particle size, in the content of cellulose, starch and vitamins were used in the treatment. The combined therapy proved to be advantageous only in the rate of the clinical effect, while the acceleration of the movement along the large intestine did not depend on the treatment type. A long-term (during one year) follow-up of the patients showed that the bran intake led to the cessation of the disease relapse; when the bran was abolished the symptoms of the disease appeared in 11 out of 12 cases. The highest effect was recorded with the bran fraction containing 55.3% cellulose, 18.3% lignin, 157 micrograms tocopherol and the lowest amount of starch--18.0%. A conclusion has been made that the wheat bran are effective in the treatment and prevention of intestinal diseases, the effectiveness of the treatment depends on the summary content of food fibers in the nutrition. PMID- 3031880 TI - [Effect of combined vitamin D and E deficiencies on calcium metabolism and bone tissue of the rat]. AB - In experiments on young rats a study was made of the influence of combined vitamin D and E deficiency on biochemical signs of rickets. It was shown that in intact animals vitamin E deficiency induced moderate hypocalcemia, a tendency to decreased Ca active transport in the small intestine and mineral saturation of the bone tissue, as well as a significantly increased Ca content in the skeletal muscles and heart. The concentration of 25-oxyvitamin D (25-OVD) in the blood serum of these animals was not changed. Combined vitamin D and E deficiency intensified disorders typical for rickets; sometimes synergy of such a negative effect was recorded. A delay in the recovery of Ca metabolism parameters, bone tissue condition and 25-OVD circulating concentration persisted, when the animals with vitamin D and E deficiency were injected cholecalciferol during 6 days. The data obtained have evidenced a possible role of vitamin E deficiency in the rickets development, that is, probably, mediated, to a certain extent, by its influence on metabolism and (or) biochemical function of vitamin D. PMID- 3031881 TI - [Low-calorie products in the Hungarian diet]. PMID- 3031882 TI - [Various aspects of the pathogenesis of the development of immunoenzyme suppression]. PMID- 3031883 TI - [Oncogenes]. AB - Within a very short time the application of molecular biology to cancer research has resulted in an essential change and extension of our knowledge of transformation processes and tumor development. For the first time these mechanisms can be understood in a causal manner and causal cancer therapy seems to be possible in the near future. In this manuscript an attempt is made to give a brief survey of the influence of oncogenes on carcinogenesis. An account is given of the origin of viral oncogenes, viral mechanisms of cell transformation and activation of cellular oncogenes. Additionally, kinetics and targets of tumor proteins are discussed. The complexity and diversity of genetic regulation of growth and differentiation is discussed in some well known diseases such as chronic myeloid leukemia, Burkitt lymphoma, retinoblastoma and acute myeloid leukemia. PMID- 3031884 TI - Genetics and human cancer: family history and common cancers. PMID- 3031885 TI - Effects of carbon tetrachloride on the liver of chickens. Early biochemical and ultrastructural alterations in the absence of detectable lipid peroxidation. AB - Administration of CCl4 i.p. to Leghorn chickens did not promote lipid peroxidation of liver microsomal lipids, as evidenced by either increased diene conjugation or by decreased arachidonic acid content. The hepatotoxin did not produce liver necrosis 24 h after dosing, but decreased the cytochrome P-450 content, and aminopyrine N-demethylase and glucose 6 phosphatase activities at 1, 3, 6 and 24 h. CCl4 administration produced dilation of the rough endoplasmic reticulum and detachment of ribosomes from their membranes. These observations suggest that lipid peroxidation is not the key event in the production of these biochemical and ultrastructural alterations, elicited by CCl4. PMID- 3031886 TI - Growth patterns of stomach cancer in relation to biological characteristics. PMID- 3031887 TI - Cytomegalovirus infection of the thyroid in immunocompromised adults. AB - Cytomegalovirus (CMV) inclusions were discovered at autopsy in the thyroid follicles in five immunocompromised adults, four of whom had acquired immune deficiency syndrome (AIDS). Although reports of viral infections of the thyroid are uncommon in adults, our experience suggests that the thyroid gland may be commonly involved in patients with disseminated CMV, and the possibility exists that CMV infection of the thyroid has the potential to cause clinical dysfunction. PMID- 3031888 TI - [Serum cathepsin activity in chronic liver diseases]. AB - Serum cathepsin activity and serum level of glycoproteins were determined in patients with chronic liver disorders (cirrhosis, chronic persistent hepatitis, chronic active hepatitis, hepatoma. An increase of cathepsin activity was found in chronic persistent hepatitis and cirrhosis of the liver. Increased level of glycoproteins was observed, the most evident in patients with chronic active hepatitis. An increase of the ratio cathepsin activity/glycoprotein concentration was found in chronic persistent hepatitis, and a decrease of this ratio was observed in chronic active hepatitis. PMID- 3031889 TI - [Current achievements in nutrition, current viewpoints in dietetics]. PMID- 3031890 TI - [Dynamics of myeloperoxidase activity and phospholipid level of neutrophils in patients with rubromycosis of the feet]. PMID- 3031891 TI - Temperature-sensitive herpes simplex virus type 1 mutants defective in the shutoff of cellular DNA synthesis and host polypeptide synthesis. AB - Two temperature-sensitive herpes simplex virus type 1 mutants, ts 1-8 and ts 199, belonging to different complementation groups, were isolated. Both mutants were defective in the shutoff of host DNA synthesis at 39.5 degrees C (nonpermissive temperature). ts 1-8 exhibited intermediate levels of viral DNA synthesis at 39.5 degrees C, while ts 199 was completely deficient in viral DNA synthesis at 39.5 degrees C. Comparative polyacrylamide gel electrophoresis of the ts 1-8, ts 199 and wild-type viral-coded polypeptides and cellular proteins produced in vivo at 34 degrees C and 39.5 degrees C during various periods post infection was performed. The results indicated that ts 1-8 and ts 199 were temperature sensitive for the secondary suppression of host polypeptide synthesis. Production of the beta (early) and gamma (late) viral polypeptides was slightly delayed in the mutant-infected cells at early times post infection at both 34 degrees C and 39.5 degrees C. This delayed protein production was not evident at later times post-infection. The ts 1-8 and ts 199 mutants were distinct from the HSV-1 viron associated host shutoff (vhs) mutants of Read and Frenkel (J. Virol. 46 (1983) 498). PMID- 3031892 TI - Detection of guinea pig cytomegalovirus nucleic acids in cultured cells with biotin-labelled hybridization probes. AB - Biotin labelled hybridization probes prepared from recombinant plasmids containing segments of the guinea pig cytomegalovirus (GPCMV) genome were used to detect GPCMV nucleic acids in guinea pig cells by in situ hybridization. The time course of GPCMV infection was assessed in two cultured cell types, guinea pig embryo (GPE) cells and 104C1 cells, a transformed and cloned guinea pig cell line. Detection of GPCMV nucleic acids was accomplished in both cell types with individual GPCMV DNA fragments and with mixtures of GPCMV DNA fragments. When compared to other established methods of GPCMV detection, the method of in situ hybridization enabled the detection of a higher percentage of positive cells early during the course of the infection. In addition, differences in the replication cycle of GPCMV in the two cultured cell lines could be demonstrated. These findings will facilitate future studies of GPCMV tissue tropism in vivo. PMID- 3031893 TI - Expression of influenza hemagglutinin-polyoma T-antigen fusion proteins in a rat embryo fibroblast cell line. AB - Plasmids encoding the amino terminal portion of an influenza virus hemagglutinin (HA) fused to polyoma virus middle T (mT) or large T (lT) sequences have been constructed. Stable expression of the chimeric proteins was obtained in established rat embryo fibroblasts following plasmid co-transfection and selection for G418 resistance. The synthesis and localization of the proteins was followed by metabolic labeling with [35S]methionine and [3H]mannose, cell fractionation, and immunoprecipitation with anti-polyoma T antibody. The HA leader and amino terminal peptide direct the synthesis of the lT and mT proteins into the endoplasmic reticulum where they undergo glycosylation, but this occurs with a very low efficiency. Most of the HA-mT and HA-lT fusion protein molecules do not enter completely into the endoplasmic reticulum, but rather achieve their normal locations in the cell as slightly higher molecular weight proteins, presumably due to the extra sequences derived from HA at their amino termini. HA mT fusion protein is found to have associated tyrosine-specific protein kinase activity precipitable with anti-src as well as anti-T antibody, and cells expressing this fusion protein have a transformed phenotype. PMID- 3031894 TI - Mammary preneoplasia and tumorigenesis in the BALB/c mouse: structure and modification of mouse mammary tumor virus DNA sequences. AB - Mouse mammary tumor virus (MMTV) DNA sequences were examined in mammary tissues from BALB/c mice, including both preneoplastic hyperplastic alveolar nodule (HAN) outgrowth lines, tumors derived from those preneoplastic tissues, and DMBA induced mammary tumors. Over 60 different HAN and tumors samples were analyzed. The D2 HAN line contained one additional provirus, whereas Cv-2 and Cv-4 HAN lines that are infected with exogenous virus exhibited multiple virus integration events. D2 tumors showed the same provirus pattern as D2 HANs, whereas Cv-2 and Cv-4 tumors exhibited a subset of the acquired proviruses found in the parental HAN populations. Differential methylation patterns of virus-specific sequences were observed that resembled those described for other tissues in which viral DNA replication and integration has occurred, i.e., acquired proviruses were hypomethylated and endogenous proviruses were methylated. In tumors that arose from HAN lines and exhibited only endogenous proviruses, demethylation of the subgenomic Mtv-6 locus was observed. Demethylation of Mtv-6 was not detected in any of the preneoplastic tissues. Altered methylation of Mtv-8 and -9 was observed in both Cv-2 and Cv-4 tumors. Finally, mammary tumors induced by DMBA carried no acquired proviruses and demethylation of endogenous MMTV proviruses was demonstrated. PMID- 3031895 TI - Feline leukemia virus envelope protein expression encoded by a recombinant vaccinia virus: apparent lack of immunogenicity in vaccinated animals. AB - We have constructed a recombinant vaccinia virus encoding the expression of the feline leukemia virus (FeLV) envelope gene of the Gardner-Arnstein strain of FeLV subgroup B. Human cells infected with the recombinant virus (vFeLVenv) express and process the FeLV envelope protein similarly to cells infected with authentic FeLV. The mature gp 70 protein is transported to and accumulates on the surface of vFeLVenv-infected cells. Vaccinia virus replication and FeLV gp70 accumulation was also observed in cells of feline and murine origin, albeit at levels somewhat reduced from those in human cells. Toward the goal of developing a recombinant vaccinia virus as a live vaccine for feline leukemia disease in cats, immunogenicity studies were performed in cats and mice. These experiments yielded surprising results: although animals mounted a typical virus-neutralizing antibody response to the vaccinia virus vector, we were unable to detect antibodies against FeLV gp70 in any of the vaccinated animals. A subsequent 'booster' immunization with killed FeLV was unable to elicit evidence of immunologic 'priming' by the recombinant virus. We are presently unable to explain the apparent lack of immunogenicity. These results may point to complexities involved in the development of vaccines to protect against retrovirus infection. PMID- 3031896 TI - [Taste aversions in the ontogeny of the rat]. AB - The characteristics of acquisition and extinction of conditioned taste aversion (CTA) were studied in rats of both sexes and various ages by pairing of saccharin solution consumption with discomfort (LiCl intoxication, rotation). Pairing of saccharin consumption with LiCl in adult rats led to acquisition of CTA, more profound in male rats than in female ones. In rats of both sexes 30 days old, a profound CTA, comparable in intensity with CTA of adult male rats was acquired. In 2 months after acquisition of CTA in adult rats its magnitude did not change, while in rats of the junior group which by that time had reached puberty, CTA was reduced in females and did not change in males. Pairing of saccharin intake and rotation led to acquisition of CTA only in rats 30 days old. The role of external factors in duration of retention of CTA acquired by pairing of saccharin solution consumption with LiCl injection was studied in male and female rats 1 and 3.5 months old. In all cases CTA was extinguished much sooner in home cages than in experimental chambers, and in the elder group sex dimorphism was found: in both situations CTA disappeared sooner in female rats. The obtained data allow to suggest modulating influences of hormonal background and of contextual stimuli on CTA acquisition and retention. PMID- 3031897 TI - [Secondary-reinforcing effects of opiate agonists in the mouse]. AB - Comparative examination of secondary-reinforcing properties of opiate agonists morphine, nalorphine, pentazocine and phencyclidine was carried out on mice using procedure of place preference conditioning in an automatic shuttle-box. Morphine, phencyclidine and pentaxocine (but not nalorphine) produced strong dose-dependent secondary-reinforcing effects. Special features of behavioural manifestations of their secondary-reinforcing action were analyzed in aspect of changes in temporal asymmetry and attendant locomotor activity of animals in the shuttle-box. On the basis of the obtained and literature data, it is suggested that mu- and sigma opiate receptors mediate secondary-reinforcing effects of opiate agonists. PMID- 3031898 TI - [Hypothesis of the information switching of conditioned reflex activity]. AB - A hypothesis of informational switching of conditioned activity is proposed on the basis of experimental data, demonstrating coding of nervous processes in distribution of uncrossed classes of neuronal interpulses intervals (IPI) and functional interaction between neurones selective to IPI. According to the hypothesis, informational switching represents the ability of nervous chains to change the channels of efferent output in response to one and the same afferent signal, depending on impulse intervals characteristics of control signal, created inside the CNS or coming from without. Schemes are given of informational switching and their description in connection with CRs switching over. PMID- 3031899 TI - [Malignant fibrous histiocytoma of the mandible in a 1 6/12-year-old boy]. AB - A report on malignant fibrous histiocytoma of the mandible in a 1 6/12-year-old boy, a condition rarely seen in children. Because of inoperability radiotherapy was used resulting in complete tumour remission. The clinical course was complicated by a tracheo-oesophageal fistula and aspiration pneumonia. The spontaneous closure of the fistula occurred 5 months after tracheostomy and catheter jejunostomy. After 2 10/12 years there is no evidence of tumour disease. Possibilities and problems of therapy are discussed. PMID- 3031900 TI - [Mediator concepts and modulation of renal compensatory adaptation]. AB - In the renal compensatory adaptation after the definition and the description of the fundamental phenomena of the functional compensation as well as of the structural adaptation is reported on mediator concepts and on modulations of the renal adaptation processes. Issuing from the central position of the sodium balance a mediator concept on natriuretic hormones (Auriculin and Endoxin) is developed which is supplemented by the renotropin mediator concept. The authors deal with the modulation of the renal compensatory adaptation (e.g. influences of age diet and so on). The pharmacotherapeutic modulation of the renal compensatory adaptation is discussed with regard to the stimulation of the tubulosecretory transport of foreign substances with para-amino hippuric acid as principal substance (including own investigations with cyclopenthiazide [Benesal]. PMID- 3031901 TI - [Quantitative calcium determination in the decalcification water in ultrasonic nitric acid decalcification of the teeth]. PMID- 3031903 TI - Modifications of cAMP in plasma and CSF after experimental brain injury. AB - A dynamic head injury produced by an unconstrained head impact was induced in 30 dogs, out of a total of 45; 15 constituted the control group. Where the energy absorbed by the dog's head is of 59,95 Joules primary brain damage is produced. If the energy absorbed is of 340.31 Joules a severe brain swelling with increase in ICP takes place, leading to the animal's death within 48-96 hours. In the group of animals with a primary brain damage there was no significant change in I.C.P. However, the concentration of cAMP in CSF rose significantly, and this rise continued for 48 hours after impact. In the group of animals with severe secondary brain damage ICP rose significantly from the first post-injury control onwards. The concentration of cAMP in CSF increases significantly after the trauma, but reaches sub-normal levels 48 hours after impact. The plasma concentration of cAMP, on the other hand, is not significantly altered in any of the three groups. The modifications of cAMP in CSF could, in our opinion constitute a good guide for the assessment of the degree of brain damage. PMID- 3031902 TI - [Preoperative (neoadjuvant) chemotherapy of small cell bronchial cancer]. AB - The high rate of locally delimited recurrences of parvicellular bronchial carcinoma, following chemotherapy and complete remission, primarily in cases of limited disease, prompted the authors to apply preoperative neoadjuvant chemotherapy to seven patients. Four of them were without tumours for six to 72 months from treatment. One patient died of myocardial infarction but with no tumour detectable 15 months after treatment. Locally delimited recurrence with additional lymphogenic spreading was recorded from one patient only. Two patients died of haematogenic metastasation. The concept should be tested for efficiency on larger groups of patients. PMID- 3031904 TI - Xylazine-induced hyperglycaemia and alpha-adrenergic receptors in sheep. PMID- 3031905 TI - [The submucous plexus (Meissner and Schabadasch) in the intestine of pigs. II. Experimental infection with coronaviruses (transmissible gastroenteritis, TGE; epizootic virus diarrhea, EVD) and rotaviruses]. PMID- 3031907 TI - The properties of Aujeszky's disease (pseudorabies) virus after serial passages in chick-embryo cell cultures in the presence of ethyl p-chlorphenoxyisobutyrate (CPIB). PMID- 3031906 TI - [Sequelae of concomitant infection of chicks with adenovirus or reovirus and the agent of avian infectious anemia (CAA)]. PMID- 3031909 TI - Cytomegalovirus hemopericarditis. Report of 1 case with histologic confirmation. AB - An otherwise healthy woman presented cytomegalovirus (CMV) infection with an haemorrhagic pericardial effusion. This disease was successfully treated by a pericardial window and corticoid administration. CMV antibody titers and observation of CMV inclusions in the pericardial biopsy (fig. 1) confirmed the infection by CMV. PMID- 3031908 TI - Changes in the spontaneous action potentials from the whole, isolated, intact bovine pituitary gland following an injection of various bovine pituitary intraglandular colloid fractions. AB - Bovine pituitary intraglandular colloid (IGC) of intermediate lobe (IL) origin with accessions from the anterior lobe (AL), can modify the spontaneous action potentials (SAP) from AL, IL and posterior lobe (PL) cells. It was discovered that intraglandular colloid contains peptides ACTH, alpha-MSH, and beta-LPH when subjected to a series of radioimmunoassays. These peptides are thought, in part, to be responsible for altering the SAP. PMID- 3031910 TI - Pleomorphic adenoma in the breast of a human female. Aspiration biopsy findings and receptor determinations. Case report. AB - A case of multiple pleomorphic adenomas ("mixed" tumour of salivary gland type) of the breast is reported. This rare benign tumour can be misinterpreted as a malignant tumour both clinically and radiologically. The aspiration biopsy findings suggested cystosarcoma phyllodes. Oestrogen and progesterone receptor determinations revealed medium high levels, comparable to carcinoma of the breast. PMID- 3031911 TI - Confirmation of the inhibitory influence of exogenous oxytocin on cortisol and ACTH in man: evidence of reproducibility. AB - The influence of low-dose oxytocin perfusion (32 mIU/min) on ACTH and cortisol plasma levels was tested in 8 normal male volunteers (age 18-26). The 1-h oxytocin perfusion periods were preceded and followed by two 1-h saline control periods. During the first period, there was a slight ACTH concentration decrease in 4 individuals. Oxytocin perfusion induced a clear-cut plasma ACTH decrease in 7 out of the volunteers, and a slight decrease in the remaining one. During the second saline infusion, there was a marked ACTH increase in 7 out of the volunteers, and a weak increase in one (P less than 0.0001). A similar pattern was observed in the plasma cortisol levels (P less than 0.00001). Furthermore, a control saline perfusion was performed in 4 of the 8 volunteers and compared to the 4 corresponding oxytocin perfusion tests: the differences between the 2 sets of tests was highly significant both for ACTH (P less than 0.003) and for cortisol (P less than 0.007). Lastly, the reproducibility of this inhibitory influence was retested in 4 volunteers: the tests were repeated under the same conditions 7 days later. There were no global differences between the 2 sets of tests either for ACTH (NS) or for cortisol (NS). ACTH and cortisol concentration fluctuations during the period between each set of tests were not significant. The following variations were measured for ACTH (NS) and for cortisol (NS). The present results confirm the inhibitory influence of low-dose oxytocin perfusion on ACTH and cortisol levels in normal males.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3031912 TI - Acquired immune deficiency syndrome (AIDS) in Japanese hemophiliacs: with special reference to a case of AIDS with inclusion body disease and Kaposi's sarcoma. PMID- 3031913 TI - Platelet thrombin receptor. PMID- 3031914 TI - The regulation of blood coagulation by the endothelium. PMID- 3031915 TI - Evolution of localization of reactions of adenosine triphosphatase (Mg++-ATP ase), 5'nucleotidase (5'nt), alkaline phosphatase (AP), and acid phosphatase (AcP) in developing rat testis. II. After CdCl2 treatment. AB - The described experiments show the influence of a single dose of cadmium chloride (1.5 mg CdCl2/kg body weight applied intraperitoneally) on development of the male gonads from the 1st d of post-fetal life to 1.5 a of life. In the case of the enzymes triggering transportation processes, adenosine triphosphatase stimulated by Mg++ (Mg++-ATP-ase), alkaline phosphatase (AP), and 5'nucleotidase (5'Nt), a considerable damages begin to appear in the 15th d of life whereas in the case of acid phosphatase (AcP) already in the 1st d of life whereas in the case of acid phosphatase (AcP) already in the 1st d of life. These damages increase with age reaching their maximum in the 45th d of life. PMID- 3031916 TI - Topogenesis of glycogen distribution pattern in post-natal rat liver in reference to the activity-rest cycle. AB - The present study, on immature rats, revealed that in the liver with exclusively diploid cell population (MD), the glycogen content was half of the adult level and the 24 h distribution pattern was reverse, i.e. highest in the evening and lowest in the morning. In these animals, in general, the protein content was high and did not show any circadian rhythm but incidentally showed a time dependent zonal distribution pattern. The low glucose-6-phosphatase activity reciprocates with low glycogen content. The relative number of cells per unit area showed a time dependent distribution pattern. The liver with equal distribution of diploid and tetraploid classes (MD:MT), attained the classical circadian rhythm of glycogen with high morning and low evening oscillation pattern. The TRIDENT measurements manifest the zonal distribution pattern with high values at the perilobular region (PL) and low at the centrolobular region (CL). The relative cell number study along the acinus demonstrated more number of cells around the PL region than that at CL region, indicating a variation in the cell size. Eventually, the protein content showed a circadian rhythm in this group, with high amount in the morning. The zonal distribution pattern always revealed the high content near the perilobular region. This could be due to more number of cells per unit area in this region. The glucose-6-phosphatase showed a circadian rhythm. The typical high glucose-6-phosphatase in the perilobular region could be further subzonated into small groups of high activity surrounded by lighter zones, thus establishing the heterogeneous function of liver parenchymal cells. PMID- 3031917 TI - Freeze-fracture replica studies of effects of ACTH treatment and hypophysectomy on the cell surface of the rat adrenal inner cortex. AB - The morphological change of the cell surface in the inner zone of the adrenal cortex was studied in intact, hypophysectomized, and ACTH-treated hypophysectomized rats, using freeze-fracture replicas. In both intact and hypophysectomized ACTH-treated animals, the P fracture face of the plasma membranes of the cortical cells facing capillaries shows groups of characteristic finger-shaped microvilli, which arise horizontally from the edges of a small cavity on the cell surface to form various configurations ranging from incompletely organized, flower bud-like to complicated, arboraceous ones. In the non-treated hypophysectomized animals, however, the microvilli in such groups decrease in number and shorten in length but increase in width, looking like a blade of a prickly-pear cactus or an irregular-shaped plate. On the other hand, presumptive exocytotic and endocytotic specializations of intra-membraneous and experimental animals. PMID- 3031918 TI - [Training of leprosy teams in health education]. PMID- 3031919 TI - [Electrophysiologic demonstration of the activity of rifampicin on nerve disease developing in mice inoculated with Hansen's bacilli in the footpad]. PMID- 3031920 TI - [Importance of neurophysiologic examinations in neural leprosy: role of the H reflex of the soleus muscle (study of 30 cases)]. PMID- 3031921 TI - [Practical problems of polychemotherapy of leprosy in the Ivory Coast]. PMID- 3031923 TI - Dental anomalies in familial adenomatous polyposis coli. AB - Forty-seven Danish and 50 Finnish patients with familial adenomatous polyposis coli (FPC) were studied by panoramic tomography (PTG) of the mandible, which showed dental abnormalities in 17% of the cases. Eleven patients (11%) had supernumerary teeth and/or compound osteomas, and nine patients (9%) had impacted permanent teeth. The frequency of the dental anomalies was statistically significantly higher in FPC patients than in the normal population; it is therefore concluded that this dental abnormality should be included in the list of extracolonic manifestations that may occur in any FPC patient. The frequency of dental anomalies was higher in Finnish than in Danish patients, probably owing to a higher frequency of extracolonic manifestations in the Finnish series. PMID- 3031922 TI - Isolation and characterization of macrophages from scrapie-infected mouse brain. AB - We have isolated and characterized a population of brain macrophages from normal and scrapie-infected mice. The cells are phagocytic, possess Fc-IgG receptors, Mac-1 surface antigen and proliferate in the presence of macrophage colony stimulating factor. They resemble microglia in that they have a plasmalemmal distribution of the enzyme nucleoside diphosphatase, a property tht is characteristic of microglia in situ. In two of the three combinations of scrapie agent and mouse strain examined, the number of brain macrophages was several fold higher than in normal control mice. The increase was not observed in mice infected intraperitoneally or in control mice inoculated with normal brain homogenate. The increase is detectable as early as 3-5 weeks postinoculation. The agent/host combination that failed to show an increase in brain macrophages is one that develops large numbers of amyloid plaques. These observations suggest that these cells are closely associated with the scrapie pathogenic process in the CNS. The failure of these cells to increase in the plaque forming model of scrapie disease also suggests that they play a role in the control of CNS amyloidogenesis. PMID- 3031924 TI - Long-term treatment with corticosteroids/ACTH in asthmatic children. IV. Skeletal maturation. AB - Skeletal maturation in severe bronchial asthma was studied in 15 children during and after treatment with depot tetracosactrin and in 6 children during treatment with prednisolone. Attained skeletal maturity before treatment was below the reference mean in the majority of children. Skeletal maturation was enhanced during treatment with depot tetracosactrin, which led to more advanced attained skeletal maturity at withdrawal than at start of treatment. There was a tendency towards larger increases of skeletal maturity than of height. The influence of adrenal androgens, released by ACTH-stimulation and good control of the disease are probably relevant factors for the acceleration of skeletal maturation. After withdrawal of depot tetracosactrin the rate of skeletal maturation normalized. During prednisolone treatment there was a delay of skeletal maturation leading to a progressive relative decrease of attained skeletal maturity, and closely related to a delay in linear growth. Treatment with depot tetracosactrin may thus induce enhancement of skeletal maturation, but with the treatment regimen found to be efficacious in bronchial asthma, the effect is not very pronounced and does not perceptibly affect the ultimate outcome of growth. PMID- 3031925 TI - Evidence for transient peripheral resistance to parathyroid hormone in premature infants. AB - Serum immunoreactive parathyroid hormone (iPTH), ionized calcium, the urinary cyclic AMP/creatinine ratio (cAMP/Cr) and some indices of bone turnover (alkaline phosphatase (AP), serum osteocalcin, and the urinary total hydroxyproline/creatinine ratio (OH-P/Cr)) were measured in 26 preterm infants during the first 4 weeks of life. Despite of stimulated parathyroid gland activity cAMP/Cr, AP, osteocalcin and OH-P/Cr were low during the first week. Thereafter iPTH decreased, whereas cAMP/Cr, and the indices of bone turnover increased, reaching high-normal values (in comparison to full-term infants) during the second and third week of life. Serum iPTH was negatively correlated to cAMP/Cr in the first week (r = -0.61, p less than 0.01), whereas the relationship became positive during the second (r = 0.47, p less than 0.05) and third (r = 0.54, p less than 0.05) week of life indicating maturation of the renal response to PTH. The study supports the concept that in premature infants a transient pseudohypoparathyroid-like state is present during the first week of life reflecting an immaturity of renal and possibly bone response to PTH. This may be an etiological factor in hypocalcemia of prematurity. PMID- 3031926 TI - Clinical epidemiology of symptomatic primary herpetic infection in children. A study of 50 cases. AB - We have studied a series of 50 children with clinical primary herpetic infection during 1975-1985. Our data confirm: absence of sex differences and seasonal variations, a peak age of incidence between 6 and 24 months, a prevalence of patients of lower social status, evidence of recurrent herpes labialis as the most frequent source of infection, and frequency of herpes simplex virus 1 gingivostomatitis. The relevant findings of this study were as follows: herpes simplex virus 2 was isolated in 10% of the patients, 6% of cases occurred in the first six months of life, infection was multifocal in 36% of cases, autoinoculation was a frequent route of transmission of genital primary infection in young children, herpetic Kaposi-Juliusberg's pustulosis in infants with atopic dermatitis was the most severe presentation of primary infection and should be more adequately prevented. PMID- 3031927 TI - Intrathecal immunoglobulin production and minor motor seizures. AB - Five infants with idiopathic infantile minor motor seizures had an elevated immunoglobulin G (IgG) index ((cerebrospinal fluid IgG/serum IgG):(cerebrospinal fluid albumin/serum albumin)). The infants had prolonged prodromal symptoms, and bad prognosis. It is suggested that immunological mechanisms may contribute to the pathophysiology in certain cases of infantile spasms. PMID- 3031928 TI - Recurrent glioblastoma treated by chemotherapy alone. PMID- 3031929 TI - Molecular characterization and comparison of plasmid content in seven different strains of Neisseria gonorrhoeae. AB - The plasmid content of one penicillin sensitive and six penicillin resistant strains of Neisseria gonorrhoeae has been examined. All strains harbour a small, phenotypically cryptic plasmid of 4.1 kilo base pairs (kb). Four of the penicillin resistant strains carry a beta-lactamase-producing plasmid of 7.3 kb. One of these also carries a large plasmid of about 40 kb. The two remaining penicillin resistant strains harbour a smaller beta-lactamase-producing plasmid of 5.5 kb. The plasmids have been subjected to digestion with a number of restriction endonucleases, and their restriction maps have been compared. Judging by the maps, the cryptic (C-) plasmids show great similarities. Except that two of them have 54 additional base-pairs (bp), and two have a HpaII site instead of a DdeI site, no differences were found. The larger beta-lactamase-producing (B-) plasmids have identical maps. The smaller seem to be homologous with the larger, except for a deletion of 1.8 kb. There is no correlation between the variant of C plasmid and type of B-plasmid harboured in the penicillin resistant strains. The evolutionary implications suggested by this finding are discussed. PMID- 3031930 TI - Mammalian muscle fiber types: comparison of excitation-contraction coupling mechanisms. AB - The mechanism of fatigue of single skeletal muscle fibers appears to involve failure or inactivation of excitation-contraction (EC) coupling. The communication between transverse tubule (TT) and sarcoplasmic reticulum (SR) membranes, whereby plasmalemma voltage controls SR Ca2+ release, may involve inositol trisphosphate (IP3) as a chemical signal. IP3 has already been shown to cause SR Ca2+ release in skeletal fast-twitch fibers; the purpose of this paper is to show that IP3 has similar effects on rabbit slow twitch fibers from soleus muscle. IP3 appears to trigger SR Ca2+ release via a mechanism which is independent of the Ca2+ induced one, since it elicits regenerative SR Ca2+ release in the presence of 20 mM procaine. IP3 also acts at a step in EC coupling beyond TT depolarization, making it a likely candidate for a TT-SR chemical signal. The possible relationship of an IP3 chemical signaling mechanism to skeletal muscle fiber fatigue and mammalian fiber type differences are discussed. PMID- 3031931 TI - Changes in cyclic 3'5'-adenosine monophosphate tissue concentration and net fluid transport in the cat's small intestine elicited by cholera toxin, arachidonic acid, vasoactive intestinal polypeptide and 5-hydroxytryptamine. AB - We have analysed tissue cyclic 3'5'-adenosine monophosphate (cAMP) concentration in different fractions of the cat's small intestinal mucosa during secretion elicited in vivo by four different secretagogues: cholera toxin (administered intraluminally), vasoactive intestinal polypeptide (VIP; given i.a.), arachidonic acid (AA; administered intraluminally) and 5-hydroxytryptamine (5-HT; given i.a.). Cholera toxin was found to increase cAMP concentration in the villi but not in the crypts. The VIP, AA and 5-HT did not influence tissue cAMP concentration despite a profuse net fluid secretion. Hexamethonium inhibited secretion elicited by cholera toxin and AA but did not significantly influence tissue cAMP concentration. There is strong evidence for the view that villus and crypt regions of the small intestinal mucosa have different functions, secretion taking place in the crypts and absorption in the villi. However, the lack of cAMP increase in the crypts reported in this study suggests that cholera toxin in this model does not reach the crypts. The results are not in agreement with a role for cAMP in mediating secretion from the crypts, but are compatible with a role of cAMP in inhibiting absorption in the villi. It is suggested that the observed fluid secretion from the crypts elicited by cholera toxin, AA and 5-HT is to a major part mediated by intramural enteric reflexes. PMID- 3031932 TI - Enhanced binding of [3H] (-) adrenaline to platelets of depressed patients with melancholia: effect of long-term clomipramine treatment. AB - The specific binding of the full agonist [3H] (-) adrenaline to platelet membranes was measured in 14 drug-free depressed patients with melancholia. There was an increased number of binding sites (Bmax) with the same apparent affinity (KD) for the radioligand, indicating that platelet alpha 2-adrenoceptor density in the high affinity state was increased in depressed patients. Long-term administration of clomipramine was associated with decreases in platelet alpha 2 adrenoceptor densities which correlated with the duration of treatment. The results support the hypothesis that in endogenous depression there is a dysfunction of the alpha 2-adrenoceptor and that antidepressant treatment might involve a time-dependent desensitization process of this receptor. PMID- 3031933 TI - Clinical observation of 3 patients with glucagonoma. PMID- 3031934 TI - Effect of naloxone on PTH and calcitonin secretion and 25-hydroxy-vitamin D concentration in chronic renal failure. PMID- 3031935 TI - Myeloperoxidase activity in normal adults and children. The population study. PMID- 3031937 TI - Morphological investigations on co-cultures of rat hepatocytes and the neonatal liver cell line RL 19. AB - Hepatocytes isolated from mature rats were cultivated together with proliferating liver cells derived from newborn rats (RL 19 line). The hepatocytes of the co cultures were viable for long period and preserved their glucose-6-phosphatase activity and glycogen content longer than the mature rat hepatocytes cultured alone. Electron microscopy revealed that the RL 19 cells were covering the hepatocytes and growing beneath them. Both the RL 19 cells and the adult hepatocytes established tight contact with each other, but between the two cell types there was always a gap bridged by microvilli and resembling Disse's space. It is assumed that the envelope formed by the RL 19 cells may have a role in the maintenance of the differentiated hepatocyte structure. PMID- 3031936 TI - Immunocytochemical investigation of ACTH-cells and vasopressin in the pituitaries of humans died from myocardial infarction. AB - The pituitaries and adrenals of 30 patients died from extensive fresh myocardial infarction and of 25 patients died from other diseases were studied. In myocardial infarction the mean weight of the above glands was significantly higher than in other diseases. In most cases an increase in number of pituitary ACTH-cells was observed with the immunoperoxidase method. In non-cardiogenic shock (another 9 cases) gland weight was also increased but without a significant increase in the number of ACTH-cells. Hyperactivity of the anterior pituitary adrenal system is due to a number of known factors. It may be assumed that patients who have infarction are either exposed to an extraordinary amount of stress stimuli or are more susceptible to stress than normal subjects. The findings may indicate the morphological basis of this situation. Of the shock phenomena, incomplete necrosis and haemorrhage of the adrenal cortex are frequent. In the pituitary neural lobe the neurosecretory material, which proved to be vasopressin with the PAP-method, was found to be increased more frequently in myocardial infarction than after other diseases. PMID- 3031938 TI - Inflammatory fibrous histiocytoma of bone. AB - An 11-year-old patient with inflammatory fibrous histiocytoma of the iliac bone is presented. In addition to routine histopathological procedures the lesion was studied by enzyme-histochemical and immuno-histochemical methods. The results of acid phosphatase, alpha-naphthyl-acetate esterase and intracytoplasmic muramidase demonstration in tumour cells supported the histiocytic nature of the presented case. PMID- 3031939 TI - Rehabilitation of patients with left ventricular dysfunction and heart failure. PMID- 3031940 TI - Increased adrenocortical responsiveness to exogenous ACTH in oral contraceptive users. AB - To evaluate the effects of changing steroid milieu on adrenocortical function, basal levels and responses of cortisol, 17-hydroxyprogesterone (17PO), androstenedione (A), dehydroepiandrosterone (DHEA), and testosterone to exogenous synthetic ACTH were investigated in six normal women during the early follicular (EF) and midluteal (ML) phases of the menstrual cycle and in five women on an oral contraceptive (OC) agent (35 micrograms ethinyl estradiol and 1 mg ethynodiol diacetate, Demulen). Baseline serum steroid and cortisol binding globulin (CBG) levels were measured on days 3-7 and 21-23 of the menstrual cycle in the normal subjects and on days 3-7 of OC treatment cycles. ACTH stimulation (10 micrograms m-2 i.v. bolus) was performed following dexamethasone suppression (0.5 mg p.o. q 6 h X 4). Basal levels of cortisol and CBG as well as cortisol responses to ACTH were increased in OC users relative to normal women tested during both the EF and ML phases of the cycle. In addition, 17PO levels were increased during the ML phase both before and following dexamethasone suppression compared to levels present in the EF phase and in OC users, no doubt because of increased ovarian steroidogenesis. PMID- 3031941 TI - Parathyroid hormone and 1.25 vitamin D3 exert opposite effects on bone. PMID- 3031942 TI - Solubilization of a guanine nucleotide-sensitive parathyroid hormone-receptor complex from canine renal cortex. PMID- 3031943 TI - Parathyroid hormone secretory responses to peroral phosphate and stimulability of serum levels of carboxyl-terminal flanking peptide (PDN-21) of the human calcitonin gene by calcium in normal subjects and osteoporotic patients. PMID- 3031944 TI - Renal adenylate cyclase stimulating action of normal and oxidized parathyroid hormone (1-34). PMID- 3031945 TI - Hypercalcemia of malignancy. PMID- 3031946 TI - Mechanism of the hypocalcemic agent WR-2721 and its acute and chronic application in eu- and hyperparathyroidism. PMID- 3031947 TI - Determinants of circulating levels of 1,25(OH)2D3 (calcitriol) in primary hyperparathyroidism (PHPT). PMID- 3031948 TI - Participation of various brain nuclei in the altered time course of genetic hypertension in SHR dependent on changes in calcium metabolism. AB - Variations in calcium alimentation influence the development of high blood pressure in genetic hypertensive rats. Different calcium feeding and changes in parathyroid hormone status cause altered cAMP-content in specific brain areas. All animals with high calcium diet show significant elevated cAMP-content in the locus coeruleus, irrespective of parathyroid status. These findings support the hypothesis that elevated cAMP-concentrations reflect an increased depressor activity of the locus coeruleus. PMID- 3031949 TI - Regulation of renal Na-K-ATPase: effects of aldosterone in phosphate depletion. PMID- 3031950 TI - Mechanisms of ion transport regulation by parathyroid hormone: cAMP/Ca2+/calmodulin and phospholipid dependent phosphorylation. PMID- 3031951 TI - Mechanism of inhibition of glycolysis by vanadate. PMID- 3031952 TI - Regulation of Na+/H+ antiport in the intact renal proximal tubular cell. PMID- 3031953 TI - Interaction of vitamin D-metabolites with adenylate cyclase/cyclic AMP system: a biological model of controlled regulation. PMID- 3031954 TI - Dibutryladenosine 3',5'-cyclic monophosphate (dBcAMP) does not mimic the action of parathyroid hormone (PTH) on canine proximal tubular basolateral membrane Na+:Ca2+. PMID- 3031955 TI - Regulation of Na-phosphate cotransport in cultured renal epithelial cells: protein-synthesis-dependent and protein-synthesis-independent pathways. PMID- 3031957 TI - The pattern of management after stroke. AB - A large population-based study of stroke is used to describe the pattern of management and the relative contribution of hospital care and community care in the year following an acute stroke episode. Two thirds of the 680 patients registered in the study were admitted to a public hospital during the acute stage of their illness, while almost one quarter of all patients were managed entirely at home. One month after the initial episode the majority of patients admitted to hospital had either died or had been discharged. Overall, four-fifths of the care during the first year after the onset of the stroke took place out of hospital, either at home or in private long-term institutions. PMID- 3031956 TI - Chronic granulomatous disease. PMID- 3031958 TI - [Anatomopathological variations in malignant tumors of the mouth]. PMID- 3031960 TI - Thanatophoric dwarfism in a Nigerian child. AB - We describe a lethal variety of congenital short-limbed dwarfism (Thanatophoric dwarfism) characterized by marked shortening of the extremities, macrocephaly with associated hydrocephalus and a narrow thorax, delivered by a 25-year-old woman in our hospital. Experience with this form of abnormality is rare and even an isolated case report will heighten awareness and assist with further management and counselling of parents. PMID- 3031959 TI - Antibody-mediated enhancement of Wesselsbron virus in P388D1 cells. AB - Antibody-mediated enhancement of Wesselsbron virus was investigated in P388D1 cell cultures. Virus infection was enhanced in culture by various dilutions of homologous and heterologous flavivirus antibody. Highest enhancement ratios and enhancing antibody titres were obtained with the homologous antibody. Enhancement of Wesselsbron virus infection in P388D1 cultures was also dependent on the multiplicity of infection (MOI) used; cultures infected at the lowest MOI produced the highest enhancement ratios. Of the four heterologous flavivirus IMAF tested for ability to enhance Wesselsbron virus infection, Potiskum virus antibody produced highest fold enhancement and possessed the highest enhancing antibody titre. Zika, Uganda S and Dakar bat IMAF produced lower fold enhancement and had lower enhancing antibody titres. PMID- 3031961 TI - Socio-economic aspects of malaria in India--a literature review. PMID- 3031962 TI - Radiological assessment of renovascular hypertension in Ibadan, Nigeria. AB - Eight cases of renovascular hypertension (RVH) encountered in the University College Hospital (UCH), Ibadan, Nigeria between 1977 and 1982 are presented and reviewed along with four additional cases, previously reported from this hospital. Seven of these cases were due to fibromuscular disease (FMD); four were due to involvement of the renal arteries by aortic aneurysms and one was due to Takayasu's arteriopathy. No case due to atherosclerosis was seen, which is in sharp contrast to the situation in the western world where atherosclerosis is the commonest cause of renal artery stenosis (RAS). Detection and management of RVH are highlighted and the current concepts and features of FMD are briefly discussed. PMID- 3031963 TI - Changes in seminal quality following varicocelectomy in infertile Nigerian males. AB - Semen analysis in 116 subfertile and infertile patients with varicocele revealed oligospermia (counts less than 20 million/ml) in 66% of patients, asthenospermia (motility of less than 50%) in 59% of patients; and teratospermia (abnormal sperm morphology of greater than 50%) in 65% of patients. The sperm motility and morphology deteriorated with decrease in sperm concentration. After varicocelectomy 53% of all patients showed improvement in the overall semen quality. The best post-operative results were noted in patients who had pre operative counts over 20 million/ml. Deterioration in semen quality was observed in 6% of all patients. Factors which possibly played a role in the response to varicocelectomy in our environment are discussed. PMID- 3031964 TI - Clostridium difficile in the normal adult faecal flora. AB - One hundred and seventy-seven out of 324 routine non-diarrhoeal faecal specimens investigated for the presence of Clostridium difficile were culture-positive. Twelve of the faecal specimens obtained from thirty-six normal healthy volunteers yielded Cl. difficile prior to oral administration of clindamycin. Thereafter all the volunteers excreted the organism from the second day to the fifth day during clindamycin administration. Three months after stoppage of oral clindamycin sixteen of the thirty-six volunteers still excreted Cl. difficile. Peak or trough serum levels of clindamycin had no relationship with the density of growth. None of the volunteers developed any Cl. difficile associated diseases. PMID- 3031965 TI - Infectivity with human ascariasis in Ibadan Oyo State, Nigeria. AB - Some of the common foodstuffs that assist in the transmission of ascariasis in Ibadan have been determined. The frequency of recovery of Ascaris ova in the different food samples examined varied from 9.6% in tomatoes to 25.6% in lettuce leaves. The indiscriminate pollution of Ibadan environment with human faeces enables the foodstuff consumed raw to serve as effective sources of heavy Ascaris infection which lead to other medical complications. The strategies for the control of Ascaris infection in Ibadan were discussed. PMID- 3031966 TI - Ascaris lumbricoides as a vehicle of bacterial infections. AB - Investigations were conducted on microflora on the body surface and internal organs of adult Ascaris lumbricoides. The investigations involved examination of wet faecal preparations of over 100 pupils using light microscope. The body surface of the adult Ascaris worms was cultured on selective media. Adult worms were dissected and different parts of the worms' gut were cultured for isolation of micro-organisms. The results of the bacteriological examination of the body surface of A. lumbricoides yielded many genera of bacterial organisms. The results from internal organs of dissected adult Ascaris worm yielded varying percentages of organisms similar to those found on the surface of the worm. The possibility of linking Ascaris infection with this aetiology of pyrexia of unknown origins as commonly seen in tropical regions was discussed. PMID- 3031967 TI - Unusual renal manifestations of choriocarcinoma. AB - The cases of two young Nigerian women, who presented with profuse haematuria and renal enlargement secondary to metastatic infiltration from choriocarcinoma in the absence of primary malignant uterine foci are reported and discussed. The rarity of this mode of presentation of choriocarcinoma is highlighted. PMID- 3031968 TI - Ectopic thoracic kidney. AB - A case of congenital thoracic kidney recently encountered at the University College Hospital, Ibadan, Nigeria is presented. Ectopic intrathoracic kidney, although very rare, should be considered in any patient with a mass at the base of the lung on a chest radiograph. Intravenous urography is diagnostic and may obviate the need for extensive investigation and operation for an asymptomatic kidney that functions normally and requires no treatment. PMID- 3031970 TI - Does exogenous magnesium suppress myocardial irritability and tachyarrhythmias in the nondigitalized patient? PMID- 3031969 TI - Radiographic evaluation of brachial plexopathy. AB - Neurologic signs and symptoms of brachial plexopathy may be subtle or confusing, making clinical localization of disease along the length of the brachial plexus difficult. To determine the most direct radiographic approach to diagnosing and anatomically delineating the cause of brachial plexopathy, we reviewed the clinical and radiographic records of 43 patients presenting with signs and symptoms referable to the brachial plexus who received CT and/or myelography as part of their radiographic evaluation. The study population was divided into two groups, those with and those without trauma. Significant deficiencies were detected in the radiographic evaluation of the nontraumatic group, with 35% of these patients having an incomplete or inappropriate CT examination that failed to visualize the full extent of the brachial plexus. In four patients, this led to a significant (greater than 6 months) delay in diagnosis. It was concluded that trauma patients presenting with brachial plexus symptoms should have cervical myelography first, rather than CT. Patients without a history of trauma should be classified on the basis of clinical findings as having central (cord, epidural space, neural foramen) or peripheral (retroclavicular space, axillary apex) disease. If the abnormality is central, myelography should be the first technique used; if peripheral disease is present, CT should be the first study. If the disease extends beyond the confines of the anatomic compartment suggested clinically, the other technique should be used for further evaluation. CT scan protocols for brachial plexus evaluation should employ bolus/drip contrast enhancement to distinguish vascular structures from masses.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3031971 TI - Experimental evaluation of technetium-99m red blood cell radionuclide ventriculography for detecting pericardial bleeding. AB - Abnormal loculated or diffuse blood pools adjacent to the heart have been observed in patients with pericardial bleeding who have been imaged by gated equilibrium radionuclide ventriculography (RNV). To study the scintigraphic appearance of fresh pericardial blood, we performed equilibrium RNV in six dogs with measured volumes (10, 30, or 50 ml) of intrapericardial blood. Loculated and diffuse pericardial blood was simulated by injecting the blood either into an intrapericardial balloon, or freely into the pericardial space. Ability to detect pericardial blood was determined by blinded review, and blood volume analysis was attempted by measuring its scintigraphic thickness, brightness (relative to the left ventricle), extent, and background-subtracted count rate and a peak count index. Detection rates for 10, 30, and 50 ml were all 100% for loculated pericardial blood, and 67%, 100% and 100% for free pericardial blood, with the use of three scintigraphic views. Visually determined "extent" of the abnormal blood pool was the most reliable indicator of pericardial blood volume. When the volume was 30 ml or more, at least 40% of the heart was surrounded in 26 of 27 cases (96%); the specificity of this finding was 90%. We conclude from this animal study that RNV should be a sensitive method for detecting pericardial bleeding; visual appearance permits qualitative assessment of the volume of accumulated labeled blood. PMID- 3031972 TI - Guidelines for vasodilator therapy of congestive heart failure in infants and children. PMID- 3031974 TI - Occupational exposure to airborne dust, respirable quartz and metals arising from refuse handling, burning and landfilling. AB - Industrial hygiene investigations were conducted in 1983 at a refuse derived fuel (RDF) burning plant, a refuse transfer station and three municipal landfill sites. The field surveys were conducted during the warmer and drier seasons of the year. The investigations included air sampling for total dust, respirable quartz and airborne metals. Bulk samples of soil cover, precipitator/boiler ash and transfer station baghouse fines were analyzed for quartz, elements, asbestos and polychlorinated biphenyls (PCBs) content. Asbestos and PCBs were not detected in any of the bulk samples taken. Quartz content of the bulk samples varied from 8% to 31%. Except for the boiler grate inspector and the precipitator cleaner at the RDF burning plant, exposure to airborne metals was not excessive at the sites tested. One personal sample (at landfill site A) for total dust exceeded 10 mg/m3 out of a total of eighteen personal and area samples. Significant exposure to respirable quartz was found at all the sites (up to 0.20 mg/m3). Respirable quartz exposures in excess of the NIOSH criteria concentration of 0.05 mg/m3 were found in three out of seven personal samples at the RDF burning plant, two out of three personal samples at the refuse transfer station and six out of nine personal samples taken at the three landfill sites. PMID- 3031973 TI - Odorization of inert gas for occupational safety: psychophysical considerations. AB - Odorization of inert gas can serve to warn workers in an enclosed space about gas leaking into the space. This psychophysical investigation, performed under conditions of directed attention, examined two candidates for possible odorization of argon:pyridine and cis-3-hexen-1-ol. Detection thresholds for pyridine and cis-3-hexen-1-ol in argon were 106 ppb and 19 ppb, respectively. Practice over four days yielded modest improvement in the detection of both odorants. For cis-3-hexen-1-ol, smokers had marginally lower thresholds than nonsmokers and older participants had slightly higher thresholds than younger participants. Gender, smoking status and age had no reliable influence on threshold for pyridine. This outcome indicated desirable perceptual stability for pyridine. Additional experiments dealt with the perceived intensity of pyridine and cis-3-hexen-1-ol over time in the realistic setting of an environmental chamber. Visitors to the chamber and occupants in the chamber assessed perceived magnitude at 5-min intervals for up to 60 min during injections of odorized argon into the chamber. Participants could gauge and track the concentration of pyridine much better than that of cis-3-hexen-1-ol. This held true for occupants almost to the same degree as visitors, though occupants inevitably exhibited some olfactory adaptation. Hence, the suprathreshold measurements also gave strong relative endorsement to pyridine. Calculations based on the experimental results indicated that odorization of the inert gas stream with 3 to 10 ppm (v/v) pyridine should suffice to warn occupants or visitors of an argon buildup of any severity. Field studies should permit a definitive judgment of the best concentration to use in practice. PMID- 3031975 TI - Respirable dust control in grinding gray iron castings. AB - High speed grinding of gray iron castings long has been associated with excessive exposure to crystalline silica. Not all workers engaged in these operations are protected by conventional ventilation techniques. Dust in the air that has been entrained by the spinning grinding wheel and not captured in the grinder hood has been postulated to be a major exposure source. A pilot grinding operation was constructed, and the size distribution and concentration of airborne particles were measured with the aerodynamic particle sizer (APS). Various control measures proved effective in reducing the respirable dust concentration: increased exhaust ventilation, and installation of baffles and/or the use of an air jet to deflect the entrained air stream. The concentration of respirable dust is the breathing zone was reduced approximately 20-fold through the combined use of increased ventilation, interior baffles, and an air jet. The air jet and baffle utilized at the base ventilation rate reduced the respirable dust concentration by a factor of three to four, whereas the baffle alone halved the concentration. PMID- 3031977 TI - Low incidence of acquired cytomegalovirus infection in neonates transfused with washed red blood cells. AB - To define the incidence of cytomegalovirus (CMV) infection in infants given transfusions of washed blood cells from random donors, 100 infants who were identified as being CMV seronegative at birth were resampled at hospital discharge and again six weeks after hospitalization. All infants received washed red blood cell products; 37 infants received nonleukodepleted platelets and/or plasma. There were 7.4 donor exposures per infant. Donor units were assayed for anti-CMV IgG and IgM at the time of donation. Seventy-six infants received at least one transfusion from a seropositive donor (mean transfusion volume, 89 mL; mean, 3.7 seropositive donor exposures). Infection was defined by seroconversion to anti-CMV. None of the recipients of exclusively seronegative blood seroconverted. A single infant who received 34 mL of washed cells from a seropositive donor (IgG+, IgM-) and 31 mL of washed cells from a seronegative donor showed IgM anti-CMV 15 days after transfusion and IgG anti-CMV at a six week follow-up visit. No recipients of IgM+ blood were infected. Our data demonstrate a 1.3% incidence of anti-CMV seroconversion following receipt of washed red cells from seropositive donors. This rate is within background levels for hospitalized neonates and is significantly lower than results of similar studies using unwashed blood. PMID- 3031976 TI - No routine role for vincristine, adriamycin, and cyclophosphamide (VAC) or thoracic radiation therapy in extensive stage small cell lung cancer. AB - A randomized trial of extensive small cell lung cancer (SMCLC) without CNS metastases comparing combination chemotherapy with the three-drug combination of vincristine, doxorubicin, and cyclophosphamide (VAC) versus a program using six drugs and starting with VAC plus Lomustine (CCNU) and etoposide (VAC-LE) was begun. After three cycles, patients without progression of disease were bronchoscoped and then randomized to the original chemotherapy program and prophylactic cranial irradiation (PCI) with or without thoracic radiation therapy (TRT). Patients with CNS metastases at diagnosis were treated with the VAC-LE program. At initial evaluation of 17 patients on VAC, and 18 without CNS metastases, and 10 with CNS metastases on VAC-LE, a statistical superiority for the VAC-LE over VAC was noted. Patients on VAC-LE (in CNS negative patients) had a higher regression rate (89 vs. 82%, 7 CR vs. 0%); a higher negative second bronchoscopy rate (69 vs. 50%); and better median (12.3 vs. 6.8 months), 2-year (14 vs. 0%), and overall survival rates (p = 0.005) than did patients on VAC. Even the VAC-LE, CNS positive patients had higher CR (20 vs. 0%), median (8.6 vs. 6.8 months), and 2-year survival rates (10 vs. 0%) than did VAC patients without CNS metastases. PMID- 3031978 TI - The importance of the virology laboratory in the diagnosis and management of viral meningitis. AB - Recent advances in cell culture techniques have made possible the rapid and accurate detection of enteroviruses, the most commonly identified cause of aseptic meningitis. Between 1983 and 1985, 69 patients were diagnosed as having enteroviral meningitis by viral culture of cerebrospinal fluid, throat swab, and/or rectal swab or stool specimens. Half of the 49 patients in whom the diagnosis was based on positive cerebrospinal fluid culture benefited directly from viral culture results by early withdrawal of antibiotics, early discharge, or changing of the diagnosis and prognosis of the disease. Enteroviral cultures became positive in as early as 24 hours, and most of the cultures became positive within one week. The cost of viral culture is comparable with that of other microbiologic tests. The virology laboratory has proved useful in the diagnosis and management of patients with enteroviral meningitis. PMID- 3031979 TI - Immunoprophylaxis of viral hepatitis: a practical guide. AB - There are presently available immune globulin preparations of proven efficacy and safety in the prophylaxis of type A and B viral hepatitis. Active vaccines for prevention of type B hepatitis are now available and are safe and effective. None of these preparations has been associated with the development of HTLV III/LAV/HIV infection. Preexposure prophylaxis for type B hepatitis is indicated for health care workers who have frequent contact with blood, and most endoscopists would be among this group of high-risk individuals. Prenatal screening of pregnant women for HBsAg provides an opportunity to intervene in the vicious cycle of perpetuating a population of hepatitis B carriers via perinatal transmission. Unfortunately until serological tags for the non-A non-B hepatitis viruses are identified, there will be no effective vaccination for this form of hepatitis. Patients who already have acute or chronic viral hepatitis must await the advent of safe and effective antiviral agents for treatment of these viral infections. PMID- 3031980 TI - Colonic small cell undifferentiated carcinoma: a distinct pathological diagnosis with therapeutic implications. AB - A case of metastatic primary small cell undifferentiated carcinoma of the colon responding to combination chemotherapy is reported. A review of small cell undifferentiated carcinoma of the gastrointestinal tract with attention to the colon is discussed. Recommendations for the management of this rare neoplasm are presented based on its similarity to the more common bronchogenic small cell undifferentiated carcinoma. PMID- 3031981 TI - Effect of IgG anti-Rho(D) in adult patients with chronic autoimmune thrombocytopenia. PMID- 3031983 TI - Neoplastic superior vena caval obstruction: diagnosis with percutaneous needle aspiration. AB - Obstruction of the superior vena cava arises from a spectrum of etiologies that include both benign and malignant conditions. Therefore, management of this serious disorder varies and depends on the underlying cause. Pursuit of a histologic diagnosis with invasive procedures has been associated with a wide range of diagnostic yields and complications. Percutaneous fine-needle aspiration biopsy has been shown to be highly reliable and well tolerated in the diagnosis of a variety of mediastinal and lung masses. Three patients are presented with obstruction of the superior vena cava in whom computed tomography safely guided percutaneous needle biopsy in obtaining a correct histologic diagnosis. It appears that transthoracic percutaneous needle aspiration biopsy is safe and efficacious in patients with superior vena cava syndrome, but further experience with this increasingly available procedure is warranted. PMID- 3031982 TI - cDNA cloning and mapping of the human creatine kinase M gene to 19q13. AB - We describe the first isolation of a human creatine kinase M cDNA clone and its mapping of the gene to human chromosome 19. A human creatine kinase M cDNA clone, pJN2CK-M, harboring a 1,160-bp insert, was isolated by colony hybridization with a previously sequenced chicken creatine kinase M cDNA probe. The human cDNA was used as a probe in Southern transfers of TaqI-digested genomic DNA from mouse/human somatic-cell hybrids to localize the human creatine kinase-M gene to chromosome 19. In situ hybridization of the tritiated cDNA probe to metaphase chromosomes of peripheral blood lymphocytes from normal males revealed significant labeling to chromosome 19. These two independent methodologies assign the human creatine kinase-M gene to chromosome 19. Since greater than 69% of the grains of chromosome 19 label band q13, the human creatine kinase-M gene has been mapped to 19q13. On the basis of high-resolution G-banding, the predominant labeling site was 19q13.2-q13.3. PMID- 3031984 TI - Prevalence, pathophysiology, and treatment of rhegmatogenous retinal detachment in treated cytomegalovirus retinitis. AB - Seventeen patients with the acquired immune deficiency syndrome and cytomegalovirus retinitis were treated with the antiviral drug ganciclovir (9 [1,3-dihydroxy-2-propoxy-methyl]-guanine, DHPG). Eight eyes of five patients developed rhegmatogenous retinal detachment after initiation of treatment. Multiple breaks in areas of peripheral, healed, atrophic retina accounted for the detachments. All seven eyes that underwent surgery had extensive retinal detachments that were reattached with vitrectomy and silicone oil. Retinotomy and retinal tacks were necessary in two cases that were complicated by severe proliferative vitreoretinopathy. In the fellow eye of one patient, laser treatment was used prophylactically to wall off a peripheral patch of healed retinitis. Endoretinal biopsies and culture were taken in five eyes; evidence of persistent cytomegalovirus was seen in two cases despite concurrent and clinically effective antiviral therapy. PMID- 3031985 TI - In vitro studies of human monocyte migration across endothelium in response to leukotriene B4 and f-Met-Leu-Phe. AB - Relatively little is known about monocyte emigration from the vasculature or about the factors that regulate this process. In this study, a human in vitro model of a blood vessel wall was used for examination of monocyte transendothelial migration. Umbilical vein endothelial cells were grown to confluency on amnion connective tissue, and human monocytes were stimulated to cross the monolayer in response to the chemoattractants leukotriene B4 or f-Met Leu-Phe. The pattern and time course of monocyte migration were similar for the two chemotactic factors. In both cases, approximately 40-50% of the adherent monocytes extended single or multiple pseudopods into the apical endothelial surface. This indenting behavior was also observed in the absence of chemotactic factors. It was not affected by the medium (M199 or Gey's) or method of monocyte isolation. Neutrophils also displayed this behavior, but only about half as many neutrophils as monocytes indented the endothelial surface. The integrity of the endothelium remained intact as the monocytes traversed the monolayer. When the monocytes reached the basal surface of the endothelium, they frequently wedged themselves between the basal surface of the endothelium and its basal lamina. The monocytes then invaded the basal lamina and accumulated in the connective tissue. In response to both f-Met-Leu-Phe and leukotriene B4, monocyte migration across the endothelium began as early as 10 minutes. The average rate of accumulation in the connective tissue peaked at 30 minutes; and by 60 minutes, 25-35% of the monocytes had traversed the monolayer. Approximately two to three times as many monocytes traversed the endothelium under conditions of chemotaxis as under conditions of chemokinesis or random migration. These studies provide the basis for understanding the process of monocyte migration out of the bloodstream and lay the foundation for the study of their differentiation into macrophages in the connective tissue. PMID- 3031988 TI - Ototoxicity of an ototopical preparation in a primate. AB - Severe ototoxicity has been observed in laboratory rodents after middle ear application of ototopical preparations or their constituents. Owing to their relatively high susceptibility, these animals may not be ideal experimental models. In the present study the ototoxicity of Cortisporin Otic Suspension (neomycin, polymyxin B, hydrocortisone) was studied using the baboon as a primate model. Middle and inner ear pathologic changes were assessed after a single application of the suspension to the tympanic cavity. Inner and outer hair cell loss occurred in all the experimental animals; it was, however, confined to the basal turn of the organ of Corti. Relative to previously studied rodents, the thicker, more densely structured round window membrane of the primate appears to provide better protection against penetration of ototoxic agents. Nonetheless, the results of this study indicate that ototopical preparations must be used with caution in clinical situations in which they may reach the middle ear. PMID- 3031986 TI - Hepatocarcinomas, cholangiocarcinomas, and hepatoblastomas produced by chemically transformed cultured rat liver epithelial cells. A light- and electron microscopic analysis. AB - The histology and ultrastructure of more than 100 tumors produced by a chemically transformed rat liver epithelial cell line and its single-cell-derived clonal subpopulations were studied. Wide-ranging morphologic presentations were observed, including carcinomas, sarcomas, "mixed epithelial-mesenchymal" tumors, and undifferentiated tumors. In addition to epidermoid and adenocarcinomas, several tumors were morphologically indistinguishable from hepatocellular carcinomas. The "mixed epithelial-mesenchymal" tumors reproduced most of the various histologic features of human hepatoblastomas. In many instances, the epithelial component occupied focal areas of tumors, and transmission electron microscopic studies of the "sarcomatous" regions revealed either spindle-shaped epithelial tumor cells or, in most cases, scattered epithelial tumor cells surrounded by numerous fibroblasts or myofibroblasts. Similar findings were observed in several "sarcomas" examined ultrastructurally, which suggests that most of the mixed tumors or sarcomas actually were spindle cell carcinomas and/or carcinomas with marked host fibroblastic reaction. However, in a few mixed tumors produced by clonally derived cell strains, unequivocal carcinosarcomas with neoplastic osteoid or chondroid tissue were demonstrated. The findings of this study are discussed in the context of current insights on the cellular composition of the liver and on the histogenesis of human hepatoblastoma. PMID- 3031987 TI - Eicosanoid production by peritoneal and splenic macrophages in mice depleted of bone marrow by 89Sr. AB - Previous studies showed that the prostaglandin-forming macrophages (M phi) induced in the spleens of CBA/J mice by intraperitoneal administration of Corynebacterium parvum (CP) could not be demonstrated following the depletion of bone marrow and blood monocytes with 89Sr. The present study compares prostaglandin E2 (PGE2), leukotriene C4 (LTC4), and LTB4 release by splenic and resident peritoneal M phi in 89Sr-treated mice and 88Sr controls following in vivo CP and in vitro incubation with zymosan, calcium ionophore A23187, or phorbol ester (PMA). Intraperitoneal administration of CP resulted in the appearance of PGE2- and LTB4-releasing M phi in the spleens of control but not 89Sr mice. The incorporation and quantitative distribution of 3H-arachidonic acid into membrane lipids, however, were comparable in test and control mice. Neither zymosan nor any of the other stimulatory agents was able to effect significant release of PGE2 in vitro. No release of LTC4 by splenic M phi was detectable under experimental or control conditions. In contrast, the capacity of resident peritoneal M phi to release PGE2, LTC4, and LTB4 was apparently unaffected by 89Sr-induced bone marrow and monocyte depletion with virtually no demonstrable elicitation. Resident peritoneal M phi removed after CP in such mice showed a dramatic decrease in PGE2 release when incubated in vitro with zymosan, A23187, or PMA. These results, taken with earlier findings, demonstrate characteristically different phenotypic expression of metabolism of certain eicosanoids by splenic M phi from the spleen and the peritoneal cavity and suggest in addition that the induction of PGE2-synthesizing M phi in the spleen by CP is dependent on either an immigrant cell originating in the bone marrow or a regulatory agent derived from a bone marrow cell. PMID- 3031989 TI - Endotoxin increases superoxide dismutase in cultured bovine pulmonary endothelial cells. AB - Manganous (Mn) and copper zinc (CuZn) superoxide dismutase (SOD) concentrations and glutathione peroxidase (GSH-Px) and catalase (CAT) activities were measured in cultured bovine pulmonary endothelial cells with and without exposure to Escherichia coli endotoxin (10(-1) micrograms/ml) over intervals of 0.5-24 h. The activities of two mitochondrial marker enzymes, fumarase and cytochrome-c oxidase, were also measured. Endotoxin exposure caused a marked increase (9-fold) in endothelial cell Mn SOD content without significant effects on GSH-Px, CAT, fumarase, or cytochrome-c oxidase activities. Endotoxin induced a slight decrease in CuZn SOD content over 24 h. This is the first report of a selective effect of endotoxin on Mn SOD in pulmonary endothelial cells. The response appears to be independent of an increase in mitochondrial activity (no change was observed in cytochrome-c oxidase or fumarase activities). These findings support the notion that endotoxin increases generation of toxic oxygen metabolites within pulmonary endothelial cells. An endotoxin-induced increase in Mn SOD could contribute to the reported protective effect of endotoxin against oxygen toxicity in these cells. PMID- 3031990 TI - Sexual dimorphism in adrenergic regulation of hepatic glycogenolysis. AB - The total phosphorylase a plus b of hepatocytes isolated from females and incubated in the absence or presence of estradiol and progesterone at concentrations found in vivo does not vary during the estrous cycle. However, there is a slight but significant influence of the estrous cycle on basal and epinephrine-stimulated phosphorylase a activity, with a nadir being seen on diestrus. The relative contributions of the alpha- and beta-mediated pathways to phosphorylase a activation do not vary with the estrous cycle but are constant at 75 and 56%, respectively, of the response to 5 X 10(-8) M epinephrine. When the epinephrine-stimulated glucose release from glycogen stores in cells from females and males is compared, the release from the female is greater than that from the male, while the alpha-receptor-mediated stimulation in the female is comparable with that in the male. The beta-mediated pathway causes glucose release which averages 45% of that stimulated by epinephrine alone in cells from females. Basal cytosolic free calcium is similar in cells from males and females (142 vs. 149 nM, respectively). The epinephrine-stimulated increase in cytostolic free calcium (Cai) is greater in the male than the female at 10(-6) M (489 vs. 380 nM) but greater in the female than the male at 5 X 10(-9) M (54 vs. 27 nM). The changes in Cai are equivalent at intermediate epinephrine concentrations. When considered with our prior analysis of 45Ca efflux after adrenergic stimulation, this suggests there may be a sexual dimorphism in hepatocyte calcium transport systems. The glucose release for a given increase in Cai is greater in the female than the male probably due to the concomitant action of the beta-mediated increase in cAMP and the alpha-mediated increase in Cai. This supports the conclusion that the beta-mediated component does make a significant contribution to the catecholamine regulation of glycogenolysis in hepatocytes from adult female rats. PMID- 3031992 TI - Hippocampal and renal type I receptors are differentially regulated. AB - Previously, we have shown that renal mineralocorticoid receptors and hippocampal "corticosterone-perferring" sites have identical intrinsic steroid specificity in vitro. Others have shown that the aldosterone binding species in kidney and hippocampus have identical trypsin fragmentation patterns on isoelectric focusing. To further explore possible areas of identity, we determined levels of type I receptors in hippocampus, renal outer medulla cortex, and renal inner medulla papilla from 22 min to 16 days after adrenalectomy. Available type I sites in kidney fractions increased postadrenalectomy to plateau levels in 22 (inner medulla papilla) or 90 min (outer medulla cortex). In contrast, available hippocampal receptors attained maximal levels 24-48 h postadrenalectomy. Animals, 24-h adrenalectomized, showed no differences in steroid uptake or washout between kidney and hippocampus, determined by in vitro tracer binding 22 or 90 min after intravenous aldosterone or corticosterone. We interpret the marked difference in receptor levels between kidney and hippocampus postadrenalectomy as evidence for tissue-specific differences in the control of receptor levels by endogenous steroids. PMID- 3031991 TI - Effects of insulin and epinephrine on Na+-K+ and glucose transport in soleus muscle. AB - To identify possible cause-effect relationships between changes in active Na+-K+ transport, resting membrane potential, and glucose transport, the effects of insulin and epinephrine were compared in rat soleus muscle. Epinephrine, which produced twice as large a hyperpolarization as insulin, induced only a modest increase in sugar transport. Ouabain, at a concentration (10(-3) M) sufficient to block active Na+-K+ transport and the hyperpolarization induced by the two hormones, did not interfere with sugar transport stimulation. After Na+ loading in K+-free buffer, the return to K+-containing standard buffer caused marked stimulation of active Na+-K+ transport, twice the hyperpolarization produced by insulin but no change in sugar transport. The insulin-induced activation of the Na+-K+ pump leads to decreased intracellular Na+ concentration and hyperpolarization, but none of these events can account for the concomitant activation of the glucose transport system. The stimulating effect of insulin on active Na+-K+ transport was not suppressed by amiloride, indicating that in intact skeletal muscle it is not elicited by a primary increase in Na+ influx via the Na+/H+-exchange system. PMID- 3031993 TI - Stimulation-associated redistribution of H+-K+-ATPase activity in isolated gastric glands. AB - The objective of this work is to establish a procedure to study the stimulation dependent membrane redistribution and properties of H+-K+-ATPase in an in vitro model system, rabbit isolated gastric glands. Stimulated (10(-4) M histamine plus 10(-5) M forskolin) and resting (10(-4) M metiamide) glands were homogenized and fractionated into PO (40 g, 5 min), P1 (400 g, 10 min), P2 (14,500 g, 10 min), P3 (48,200 g, 90 min), and supernatant, S3. Significant changes occurred in the distribution of our marker for H+-K+-ATPase (K+-p-nitrophenyl phosphatase) activity: a reduction in activity of P3 and a compensatory increment in P1. P3 showed valinomycin (Val)-dependent vesicular H+ uptake, while H+ uptake in P1 was Val independent. Direct measurements of ATPase revealed that H+-K+-ATPase activity of P3 was Val dependent and decreased by stimulation; H+-K+-ATPase activity of P1 was Val independent and increased by stimulation. Further density gradient purification of P1 showed that membranes lighter than 17% Ficoll contained higher specific H+-K+-ATPase activity, and the observed increase in H+ K+-ATPase associated with stimulation was more pronounced. Also, the lighter fractions from stimulated P1 had much latent H+-K+-ATPase activity that was unmasked by n-octylglucoside. The properties of membrane fractions from isolated glands were consistent with results obtained in vivo: high H+-K+-ATPase activity of P3 from resting glands corresponds to cytoplasmic tubulovesicles lacking KCl transport pathways; high activity of P1 from stimulated glands corresponds to apical plasma membrane vesicles containing KCl transport in addition to the H+-K+ ATPase, and full competency for the generation of HCl. PMID- 3031994 TI - Phorbol esters and A23187 regulate Na+-K+-pump activity in pancreatic acinar cells. AB - To clarify the subcellular mechanisms that mediate stimulation of Na+-K+-pump activity in pancreatic acinar cells by cholinergic agonists, we examined the effects of the phorbol ester, 12-O-tetradecanoylphorbol-13-acetate (TPA) and the Ca2+ ionophore A23187 on [3H]ouabain binding to dispersed guinea pig pancreatic acinar cells under conditions in which binding reflects the average rate of pump cycling. The phorbol ester more than doubled Na+-K+-pump activity as did the diacylglycerol analogue, 1-oleoyl-2-acetolyl-sn-3-glycerol. A23187 increased pump activity by a maximum of 31% at 0.3 microM but was progressively inhibitory at higher concentrations. The stimulatory effects of TPA and A23187 were additive, although either secretagogue elicited a less than additive response when added together with a maximally effective concentration of the cholinergic agonist, carbachol. Removal of extracellular Ca2+ had little effect on the pump response to TPA and did not reduce the maximal effect of A23187 but abolished the inhibitory effect seen at high ionophore concentrations in Ca2+-containing medium. These results indicate that both Ca2+ and protein kinase c are involved in regulating Na+-K+-pump activity in the pancreatic acinar cell. PMID- 3031995 TI - Mechanisms of active Cl- secretion by frog gastric mucosa. AB - Net Cl- flux across the bullfrog gastric mucosa was examined to test the hypothesis that Cl-secretion (JClnet) can be driven by either of the two cation exchange pumps in the oxyntic cell. The effects on JClnet of ouabain, an Na+-K+ pump inhibitor, and omeprazole, an H+-K+ pump inhibitor were examined. Omeprazole abolished acid secretion (JH) and reduced JClnet in bullfrog gastric mucosa. For mucosae at open circuit the omeprazole-induced decrease in JH was not significantly different than the decrease in JClnet, and the transmucosal potential difference (PD) was increased. When short-circuited mucosae were treated with omeprazole, the decrease in JClnet was significantly less than the decrease in JH, and short-circuit current (SCC) was correspondingly increased. After treatment of short-circuited mucosae with ouabain, the omeprazole-induced decreases in JH and JClnet were not significantly different, and no change in SCC occurred. For open-circuited mucosae, pretreatment with ouabain resulted in a significantly smaller omeprazole-induced increase in the transmucosal PD than was seen without ouabain pretreatment. Our data 1) show that both the H+-K+ pump and the Na+-K+ pump can drive Cl- secretion and 2) suggest that inhibition of the H+ K+ pump with omeprazole stimulates the Na+-K+ pump. PMID- 3031997 TI - Effects of parathyroid hormone on net proton flux from neonatal mouse calvariae. AB - Bone mineral buffers protons during acute metabolic acidosis; whether parathyroid hormone (PTH) augments proton buffering is controversial. To determine whether PTH augments proton buffering by bone, we cultured neonatal mouse calvariae with or without PTH (10(-8) M) for 3 h in medium that was physiologically acid (pH approximately 7.20), neutral (pH approximately 7.40), or alkaline (pH approximately 7.60). Over the entire pH range studied there was less influx of protons into calvariae treated with PTH than into control calvariae, indicating that PTH does not augment but instead inhibits proton buffering by bone. To determine whether chronic exposure to PTH is necessary to augment proton buffering, calvariae were incubated with PTH for 24 h before a 3-h culture. Calcium efflux from calvariae exposed to PTH (10(-8) M) for 24 h exceeded that of controls. When these same calvariae were recultured for 3 h in fresh medium, PTH treated and control calvariae behaved similarly, with net efflux of protons into acid, neutral, and alkaline media. Regardless of whether PTH is added at the time of exposure to acid medium or 24 h before calvariae cultured with PTH do not buffer protons to a greater extent than controls. PMID- 3031996 TI - Dihydroxy bile salt-induced secretion of rubidium ion across the rabbit distal colon. AB - Possible mechanisms of dihydroxy bile salt-induced K+ secretion by the mammalian colon were evaluated by studying the effects of taurochenodeoxycholate (TCDC) on 86Rb+ transport across the isolated, short-circuited rabbit distal colon. Simultaneous measurements of 86Rb+ and 42K+ unidirectional fluxes were highly correlated [r = 0.964 for serosal (s) to mucosal (m) and 0.765 for m to s], indicating that Rb+ is a suitable tracer for K+ transport across the colon. Furthermore, mucosal Ba2+ (4 mM) or serosal ouabain (0.1 mM) decreased serosal to mucosal rubidium flux (JRbs----m) (from 0.24 +/- 0.02 to 0.09 +/- 0.02, and 0.08 +/- 0.01 mu eq X h-1 X cm-2, respectively) without affecting JRbm----s. Dibutyryl cyclic adenosine monophosphate (dBcAMP, 0.5 mM serosal) specifically increased JRbs----m of controls (from 0.21 +/- 0.05 to 0.67 +/- 0.09 mu eq X h-1 X cm-2) through a barium- (4 mM, mucosal) sensitive pathway without affecting JRbs----m. Mucosal addition of 2 mM TCDC increased tissue conductance (GT), reduced short circuit (Isc) slightly, and reversed JRbnet (from 0.13 +/- 0.05 to -0.29 +/- 0.08 mu eq X h-1 X cm-2) principally by increasing JRbs----m. The TCDC-induced increases in JRbs----m were reduced by 0.1 mM serosal ouabain (from 0.53 +/- 0.03 to 0.11 +/- 0.02 mu eq X h-1 X cm-2) or 4 mM mucosal Ba2+ (from 0.76 +/- 0.07 to 0.32 +/- 0.04 mu eq X h-1 X cm-2).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3031998 TI - Conductive Na+ transport in an epithelial cell line (LLC-PK1) with characteristics of proximal tubular cells. AB - Na+ influx and efflux from confluent monolayers of an epithelial cell line with multiple differentiated characteristics of the straight segment of the renal proximal tubule were studied in the presence and absence of a pH gradient. The results show that Na+ influx in the absence of a pH gradient is inhibited by amiloride as well as by complete replacement of Cl- by an impermeable anion, such as isethionate. Dissipation of cell membrane potential by increasing the potassium concentration of the extracellular medium in the presence of valinomycin also inhibited Na+ influx, whereas sodium influx induced by an H+ gradient was not affected. Inhibition of Na+ influx by different maneuvers produced hyperpolarization of the plasma cell membrane, as would be expected if the sodium movement involved net displacement of charges. Calcium and other divalent and trivalent cations also inhibited Na+ influx measured in the absence of an H+ gradient. Na+ influx induced by a pH gradient, however, was not affected. Like the Na+-H+-exchange system, the conductive Na+ pathway is localized in the apical membrane of the epithelial cells. From these results, we conclude that at least a fraction of transepithelial Na+ transport in LLC-PK1 monolayers occurs through a simple rheogenic transport system. PMID- 3031999 TI - Transepithelial and intracellular potentials in isolated Malpighian tubules of tenebrionid beetle. AB - Malpighian tubules of Onymacris plana (Coleoptera: Tenebrionidae) have been isolated for measurement of transepithelial and intracellular potentials, before and during stimulation of fluid secretion. In a bathing medium resembling the hemolymph composition of the insect, the transepithelial potential (VT) was approximately 13 mV, lumen positive. VT was subject to drift and frequently showed super-imposed regular oscillations, which were apparently action potentials associated with contractions of muscle fibers running along the tubules. Although tubules of Onymacris are approximately 8 cm long, the basal membrane potential (Vb) did not vary with distance along the tubule, averaging 31 mV. Addition of adenosine 3',5'-cyclic monophosphate (cAMP) or diuretic hormone (DH) homogenate to the bathing medium had no effect on Vb, but opposing effects on VT: cAMP caused it to increase to 60 mV, whereas DH homogenate caused a rapid drop in VT to almost zero. Ion substitutions in the bathing medium showed that under control conditions beetle tubules possessed appreciable basal permeability to both K and Cl ions, with a 10-fold reduction in bath K concentration hyperpolarizing Vb by 54 mV. The basal K and Cl channels were partially blocked by barium and thiocyanate ions, respectively. Stimulation with cAMP increased the apical membrane potential (Va) and significantly reduced the Cl permeability of the basal membrane, whereas its Na permeability remained negligible. PMID- 3032001 TI - Glucocorticoid inhibition of neurohypophysial vasopressin secretion. AB - Several lines of evidence have suggested that neurohypophysial vasopressin secretion is under the influence of glucocorticoid negative feedback. Studies in clinical and experimental adrenal insufficiency have suggested that the impaired water excretion accompanying that syndrome may be due to elevated vasopressin levels. Furthermore, both the impaired water excretion and elevated vasopressin levels observed in adrenal insufficiency may be normalized by glucocorticoid treatment. This topic remains controversial, with a considerable body of evidence suggesting that vasopressin is elevated during adrenal insufficiency not because of a loss of central steroid negative feedback but because of alterations in plasma volume osmolality (renal mechanisms). Vasopressin responses to a variety of stimuli (hemorrhage, hypoxia, hypertonic saline) in normal humans and animals appear to be attenuated or eliminated by pretreatment with glucocorticoids. However, the vasopressinergic system appears to be considerably less sensitive to negative feedback than the corticotropin-releasing factor-adrenocorticotropic hormone (ACTH) system. There is evidence that the locus for this inhibitory effect is both directly at the posterior pituitary and within the hypothalamus. It is unlikely that corticosteroid negative feedback closes a direct hypothalamo neurohypophysial-adrenocortical feedback loop. Since neurohypophysial vasopressin is involved in the control of ACTH secretion, it is more likely that the modulation of neurohypophysial vasopressin by glucocorticoid is an integral part of the overall negative-feedback control of ACTH secretion. The physiological role of glucocorticoid inhibition of vasopressin secretion remains speculative. PMID- 3032000 TI - Electroneutral H+ secretion in distal tubule of Amphiuma. AB - This study examines the cellular mechanisms of acid secretion by the in vitro perfused late distal tubule of Amphiuma kidney. Acidification of tubule fluid occurred against an electrochemical gradient of 16 mV; thus H+ secretion was active. Amiloride (1 mM) or a reduction of sodium in the perfusion fluid (from 83.7 to 7.7 mM) partially reduced acidification. Amiloride, in the presence of low sodium, completely inhibited acidification. Furthermore, acetazolamide and ouabain in the bath solution (0.1 mM) also inhibited acidification. Conductive properties of the epithelium and of individual cell membranes were determined by means of cable analysis of the tubule and intracellular voltage recordings. The transepithelial voltage and resistance averaged -0.4 +/- 0.4 mV, lumen negative, and 7,147 +/- 845 omega X cm, respectively. Two functionally different cell types were identified by intracellular microelectrodes. Type I cells had a basolateral membrane voltage (Vbl) of -67.7 mV. As determined by ion substitution experiments, the basolateral membrane was conductive to K+ and Cl-. This cell also had a 4-acetamido-4'-isothiocyanostilbene-2-2'-disulfonic acid (SITS) sensitive Na+-dependent HCO3- exit pathway in the basolateral membrane. Type II cells had a Vbl of -76.1 mV (P less than 0.05 vs. type I) and the basolateral membrane was conductive to K+ and Cl- but not to HCO3-. HCO3- movement across the basolateral membrane in this cell may occur by electroneutral Cl- -HCO3- exchange. The apical cell membrane of both cell types did not contain measurable ionic conductances, as evidenced by a high value of apical membrane fractional resistance (0.98 +/- 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3032002 TI - Forebrain contribution to sympathetic nerve discharge in anesthetized cats. AB - We tested the hypothesis that the forebrain is responsible for a component of basal sympathetic nerve discharge (SND) in the anesthetized cat. For this purpose integrated postganglionic inferior cardiac SND and blood pressure were measured before and after serial transections of the midbrain at stereotaxic planes A3 and AP0. In the first series of experiments on baroreceptor-denervated cats anesthetized with alpha-chloralose, A3 transection significantly reduced SND to 62 +/- 7% of control. Blood pressure was reduced 33 +/- 4 mmHg. SND and frontal parietal cortical activity were temporally related prior to A3 transection in many of these animals. Recovery of SND and blood pressure to control levels occurred within 30 min after A3 transection. A second midbrain transection at AP0 failed to affect SND and blood pressure at this time. This observation suggests that the effects of A3 transection were due to the loss of a component of SND of forebrain origin rather than to generalized trauma. The effects of A3 transection were not attributable to cataleptic anesthesia with alpha-chloralose, since SND and blood pressure were significantly reduced by midbrain transection in baroreceptor-denervated cats anesthetized with diallylbarbiturate-urethan. A3 transection also reduced SND and blood pressure in some baroreceptor-innervated cats. Finally, medial diencephalic ablation prevented the effects of A3 transection. We conclude that the forebrain is responsible for a significant component of sympathetic tone in anesthetized cats. Recovery from the loss of this component, however, occurs rapidly even in animals deprived of their compensatory baroreceptor reflex mechanisms. PMID- 3032003 TI - Hormonal regulation of hepatic glycogenolysis in the carp, Cyprinus carpio. AB - Carp (Cyprinus carpio) liver maintained normal glycogen content and enzyme complement for several days in organ culture. Epinephrine-stimulated glycogenolysis, phosphorylase activation, and cyclic AMP (cAMP) accumulation in a concentration-dependent manner with EC50s of 100, 100, and 500 nM, respectively. These actions were blocked by the beta-adrenergic antagonist, propranolol, but not by the alpha-adrenergic antagonist phentolamine. Glycogenolysis and tissue cAMP were uninfluenced by 10(-6) M arginine vasotocin, arginine vasopressin, lysine vasotocin, lysine vasopressin, mesotocin, or oxytocin, but were slightly increased by 10(-5) M isotocin and slightly decreased by 10(-6) M angiotensin II. [125I]-iodocyanopindolol (ICP), a beta-adrenergic ligand, bound to isolated carp liver membranes with a KD of 83 pM. Maximum binding of 45 fmol/mg protein was at 600 pM. Propranolol, isoprenaline, epinephrine, phenylephrine, norepinephrine, and phenoxybenzamine displaced ICP with KDs of 100 nM, 2, 20, 20, 60, and 200 microM, respectively. The alpha-adrenergic antagonists, yohimbine and prazosin, showed no specific binding. These data provide evidence that catecholamines act via beta-adrenergic receptors in carp liver and that alpha-adrenergic receptors are not present. Vasoactive peptides play no significant role in regulation of carp liver glycogenolysis. PMID- 3032004 TI - Renal brush-border Na+-H+ exchange activity in the aging rat. AB - Amiloride-sensitive Na+-H+ exchange activity in brush-border membrane vesicles isolated from male rat proximal tubules was decreased in the senescent rat (24 mo) compared with the young adult (6 mo). There was no significant loss in Na+-H+ exchange activity in the kidneys of animals between 6 and 18 mo of age. Amiloride insensitive Na+ uptake and the rate of pH gradient dissipation were not altered during aging. The decrease in sodium-dependent phosphate transport preceded the decline in Na+-H+ exchange activity by at least 6 mo. Sodium-dependent glucose transport was not significantly altered during aging. Thus various renal plasma membrane transport functions were affected differently in the aging rat. The decrease in Na+-H+ exchange activity during aging contrasted with the increase in exchange activity reported previously in acute ablation models of chronic renal failure. PMID- 3032005 TI - Arterial baroreceptor and vagal inputs to sympathoexcitatory neurons in rat medulla. AB - Lumbar sympathetic nerve discharge and unit activity of reticulospinal sympathoexcitatory (SE) neurons located in nucleus paragigantocellularis lateralis (PGCL) were recorded in rats. The sympathoinhibition produced by low frequency stimulation of vagal afferents was abolished by bilateral microinjections of 20 pmol of bicuculline methiodide (BIC, a GABA-receptor antagonist) into PGCL and was converted into a pressor response by injections of 100 pmol. These BIC injections also inhibited the arterial baroreflex in a dose dependent manner. In contrast the sympathoexcitation produced by high-frequency stimulation of vagal afferents was selectively blocked by bilateral injections of kynurenic acid (KYN, a Glu-receptor antagonist) into PGCL. Convergence of vagal excitatory, vagal inhibitory, and arterial baroreceptor inputs was detected in all SE neurons recorded. Single-pulse stimulation of vagal afferents produced up to two peaks of excitation of SE neurons, both blocked by iontophoretic applications of KYN and at least one inhibitory period selectively blocked by iontophoresis of BIC. The results emphasize the importance of SE neurons and surrounding area in integrating the brain vasomotor output to spinal preganglionic neurons. PMID- 3032006 TI - Quantitation of phosphorus excretion in sheep by compartmental analysis. AB - The control of phosphorus excretion in sheep has been examined by constructing a kinetic model that contains a mechanistic set of connections between blood and gastrointestinal tract. The model was developed using experimental data from chaff-fed sheep and gives an accurate description of the absorption and excretion of phosphorus in feces and urine of the ruminating sheep. Simulation of the response to an intravenous phosphorus infusion by adding an inflow of 2 g/day of phosphorus into the compartment describing blood, predicted values for fecal output of phosphorus lower than found experimentally. However, by alteration of the parameters describing absorption or salivation, the predictions approached experimental values. Similarly simulation of the conditions existing when a liquid diet was infused directly into the abomasum, i.e., a decrease in salivation rate [L(4.1)] and dietary phosphorus entering compartment 5 (abomasum) instead of compartment 4 (rumen), gave incorrect predictions for plasma and urinary phosphorus, but when the parameter for urinary phosphorus was increased the predicted values approached experimental values. These results indicated the main control site for phosphorus excretion in the ruminating sheep was the gastrointestinal tract, whereas for the nonruminating sheep fed the liquid diet, control was exerted by the kidney. A critical factor in the induction of adaptation of phosphorus reabsorption by the kidney was the reduction in salivation, and since this response occurred independently of marked changes in the delivery of phosphorus to the kidney, a humoral factor may be involved in this communication between salivary gland and kidney. PMID- 3032007 TI - Negative-feedback inhibition of fetal ACTH secretion by maternal cortisol. AB - Previous studies in this laboratory have shown that small increases in fetal cortisol (F) decreased basal fetal plasma renin activity (PRA) and completely inhibited the fetal adrenocorticotropin hormone (ACTH) response to hypotension. The present study was designed to quantitate suppression of fetal ACTH and renin secretion by maternal F. Fetal and maternal femoral arteries and veins were chronically catheterized (11 fetuses, 118-129 days gestation). Maternal intravenous infusion of 0, 0.5, 1.0, and 2.0 micrograms F X kg-1 X min-1 (n = 5 6) increased mean maternal and fetal cortisol and suppressed fetal ACTH responses to a subsequent period of hypotension in a dose-related manner. Increases in fetal plasma cortisol to 8.3 ng/ml completely suppressed the fetal ACTH response to hypotension. The results indicate that increases in maternal plasma cortisol concentration sufficient to produce modest increases in fetal plasma cortisol inhibit fetal ACTH secretion. PMID- 3032009 TI - Natural killer cell cytotoxicity and antibody-dependent cellular cytotoxicity to herpes simplex virus-infected cells in human pregnancy. AB - Natural killer cell (NKC) cytotoxicity and antibody-dependent cellular cytotoxicity (ADCC) represent the ability of human leukocyte effector cells to destroy target cells in the absence and presence of antibody, respectively. Since these immune systems play a pivotal role in the body's primary lines of defense against a variety of pathogens including herpes simplex virus (HSV), a study was undertaken to evaluate the influence of pregnancy on these systems. Eleven uncomplicated gravidas were followed serially through each trimester and compared to 11 nonpregnant female controls. Mononuclear cells were acquired by Ficoll Hypaque centrifugation of heparinized blood. Chang liver cells infected with HSV I were utilized as target cells in a 51Cr release assay. Mean NKC values in the pregnant patients were uniformly lower than in the controls. No similar decreases in ADCC activity were observed in a comparison between the two study populations. These data support previous observations suggesting that pregnancy represents a relatively immunocompromised state. Differences apparently exist between NKC and ADCC effector cell populations with regard to the influence of pregnancy. Although these physiologic alterations in immunoregulation may help support the fetoplacental allograph, detrimental conditions may exist regarding susceptibility to various pathogens such as HSV. PMID- 3032008 TI - Vasopressin release in response to nausea-producing agents and cholecystokinin in monkeys. AB - Administration of lithium chloride and copper sulfate to adult monkeys caused marked elevations in plasma vasopressin (AVP) levels without significant increases in plasma oxytocin (OT) levels. Emesis was produced in five of the seven animals given these agents, in support of nausea as the main stimulus to AVP release. A similar pattern of AVP release without OT release was found after administration of cholecystokinin (CCK). Although most monkeys vomited in response to 10 micrograms/kg of CCK, a significant increase in plasma AVP levels also was produced with a dose of 1 microgram/kg, which did not produce emesis in any animal. These findings are in marked contrast with previous results in rats, which indicated that lithium chloride, copper sulfate, and CCK each stimulated OT rather than AVP release. Despite this interspecies difference, the significant neurohypophysial hormone secretion in response to both nausea-producing agents and CCK suggests that AVP secretion in monkeys, similar to OT secretion in rats, might reflect activation of central pathways mediating nausea and/or inhibition of food intake, even when overt illness is not produced. PMID- 3032011 TI - Mammary and extramammary Paget's disease. PMID- 3032010 TI - Synaptophysin. A new marker for pancreatic neuroendocrine tumors. AB - Synaptophysin (SYP) is a glycoprotein recently isolated from presynaptic vesicles of bovine neurons. Initial studies have demonstrated its presence in neurons in the brain, spinal cord and retina, and in adrenal medullary cells. A subsequent study demonstrated it in pancreatic islet cells and certain neuroendocrine (NE) neoplasms, including several pancreatic islet cell tumors. Based on these preliminary observations, we examined, by immunohistochemistry, conventionally fixed, paraffin sections of 57 pancreatic endocrine tumors with a monoclonal antibody to SYP. Furthermore, we compared the SYP immunoreactivity of 30 of these same tumors with that of neuron-specific enolase (NSE) and of chromogranin (CG). SYP was demonstrated in all but one of the 57 tumors. In the comparative study, for which material was available in only 30 cases, SYP and NSE were present in 29 of the tumors, whereas CG was seen in only 15 cases. We conclude that SYP is a highly sensitive and useful marker for pancreatic NE neoplasms. Moreover, in view of the increasingly evident limited specificity of NSE, SYP should be considered the marker of choice for pancreatic NE neoplasms. PMID- 3032012 TI - Essential fatty acids, prostaglandins, and alcoholism: an overview. AB - Essential fatty acids (EFAs) are major structural components of the brain and through their effects on membrane properties can influence nerve conduction, transmitter release, and transmitter action. Prostaglandins (PGs) derived from EFAs have profound behavioral effects and are also able to modify conduction and transmitter function. Effects of alcohol on EFAs and PGs are therefore good candidates for explaining at least some of the actions of alcohol on brain function. Ethanol has three main known actions on EFA and PG metabolism: it reduces blood linoleic acid levels and induces or exaggerates EFA deficiency states; it blocks metabolism of linoleic acid to EFA metabolites which are known to be important in brain structure; and it enhances conversion of the linoleic acid metabolite, dihomo-gamma-linolenic acid, to PGE1. This review demonstrates that some of the short-term behavioral effects of ethanol and some of its long term adverse effects on brain, liver, and other tissues may be partly explicable in terms of ethanol actions on EFA and PG metabolism. Modification of such metabolism by dietary and other means has already been shown to influence the effects of alcohol and alcohol withdrawal in both humans and animals. This promises to be a fruitful source of investigation with substantial implications for the understanding and treatment of alcoholism. PMID- 3032014 TI - Effects of habitual alcohol intake and cigarette smoking on the development of hepatocellular carcinoma. AB - To study the effects of habitual alcohol intake and cigarette smoking on the latency period for the development of hepatocellular carcinoma (HCC), 455 patients with HCC were analyzed with respect to age at diagnosis. They were divided into hepatitis B virus (HBV) positive and negative patients based on HbsAg and high titer anti-HBc in serum. HBV-positive and negative HCC patients were further subdivided into four subgroups based on the history of drinking more than one small bottle of Japanese "sake" or its equivalent per day for more than 10 yr and the history of smoking more than one cigarette per day for more than 10 yr. Among HBV-positive HCC patients, the average age of those with a drinking and a smoking habit (50 +/- 10 yr) was younger compared with that of patients with a drinking habit but without a smoking habit (56 +/- 14 yr, not significant, NS), of those who were smokers and nondrinkers (55 +/- 10 yr, NS) and of those who did not drink nor smoke (59 +/- 8 yr, p less than 0.005). Among HBV-negative HCC patients, patients with drinking and smoking habits (57 +/- 9 yr) were younger compared with those with a drinking habit without smoking (59 +/- 9 yr, NS), those who were smokers and nondrinkers (62 +/- 9 yr, p less than 0.005), and those who were nondrinkers and nonsmokers (63 +/- 12 yr, p less than 0.005). These data suggest that habitual alcohol intake may promote the development of HCC if the patients smoke cigarette regardless of the status of HBV seromakers. PMID- 3032013 TI - Hepatic triacylglycerol accumulation induced by ethanol and carbon tetrachloride: interactions with essential fatty acids and prostaglandins. AB - Triacylglycerol accumulation in the liver (fatty liver) caused by ethanol or carbon tetrachloride involves interactions with essential fatty acids and prostaglandins. The degree to which the fatty liver develops is dependent on total dietary fat intake. Both ethanol and carbon tetrachloride impair desaturation of linoleic acid and dihomo-gamma-linolenic acid and this appears to be relevant to the pathogenesis of fatty liver from two points of view. First, low arachidonic acid in liver phospholipids is associated with increased liver triacylglycerol content whether caused by ethanol, carbon tetrachloride, or essential fatty acid deficiency. Second, essential fatty acids including gamma linolenic acid and arachidonic acid, as well as the prostaglandins, prevent ethanol- and carbon tetrachloride-induced fatty liver. Arachidonic acid and possibly the prostaglandins are therefore likely to be directly involved in lipoprotein and triacylglycerol secretion by the liver. PMID- 3032015 TI - Opiate receptors: an introduction. AB - Current status of opiate receptors and their agonists is reviewed--basic aspects of receptor theory, the importance of stereospecificity in drug-receptor interactions and the role of 'second messengers' in drug action. The three classes of endogenous opioids, originating from three distinct genes, are discussed: pro-opiomelanocortin, giving rise to beta-endorphin, ACTH and various MSHs; pro-enkephalin, giving methionine enkephalin and leucine enkephalin; and prodynorphin; their anatomical distribution and the main classes of receptors with which they interact, the mu-receptor, with a high affinity for met enkephalin and beta-endorphin (as well as morphine and dynorphin A); the delta receptor for which the primary ligand is leu-enkephalin; and the kappa-receptor which is the main target for the dynorphins. Functional roles for endogenous opioids are considered. Essentially they are inhibitory to target neurones, depressing motor reflexes, baroreflexes and nociception. They also have roles in the response to physical and psychological stress. PMID- 3032016 TI - High-performance liquid chromatography for the purification of restriction endonucleases, application to BanII, SacI, and SphI. AB - Conventional fractionation methods are time consuming, thus they prolong the time required to process low-stability restriction enzymes. We now report a rapid and effective two-step chromatographic method that affords high purity endonucleases in a short time. Accordingly, an inexpensive chromatographic adsorbent such as phosphocellulose or dyed agarose in the first step is coupled to a high performance ion exchanger, namely, MonoQ, in the second step. The purification schemes reported here are now in routine use to prepare high-purity BanII, SacI, and SphI as judged by the "overdigestion" and "cut-ligate-recut" stringent quality tests. PMID- 3032017 TI - Pholasin--a bioluminescent indicator for detecting activation of single neutrophils. AB - Pholasin is the protein-bound luciferin from the bivalve mollusc Pholas dactylus which reacts with its luciferase and molecular oxygen to produce light. Pholasin was purified 226-fold with a yield of 58% from P. dactylus to give a preparation free from luciferase contamination. The ratio (k) of endogenous pholasin chemiluminescence to that when maximally stimulated by luciferase was 8.12 X 10( 6) +/- 0.87 X 10(-6) (mean +/- SD, n = 6), equivalent to a t 1/2 of 23.7 h at pH 9. Pholasin could detect reactive oxygen metabolite production from neutrophils stimulated by the chemotactic peptide N-formyl-Met-Leu-Phe, in the presence and absence of 2-chloroadenosine or cytochalasin B, and by latex beads in the presence and absence of cytochalasin B. Pholasin was also able to detect a longer lived oxidative activity distinct from myeloperoxidase, and released from neutrophils activated by latex beads or chemotactic peptide; luminol could not. Under optimal conditions pholasin produced a signal some 50-100 times that of luminol in the presence of activated neutrophils. This enabled activation of a single neutrophil by chemotactic peptide (1 microM) to be detected, giving a signal of 194 +/- 21 chemiluminescent counts per second, some six times that of the background signal (mean +/- SD, n = 2). Pholasin thus provides an indicator sufficiently sensitive to establish whether neutrophil activation occurs through thresholds in individual cells. PMID- 3032018 TI - A simple and rapid assay for heme attachment to apocytochrome c. AB - A method for assaying the covalent attachment of heme to apoprotein of cytochrome c was developed. 125I-labeled apoprotein was chemically prepared from 125I labeled yeast cytochrome c (iso-1-cytochrome c). After incubation of 125I apocytochrome c with yeast mitochondria, the product was extracted with Triton X 100, digested with trypsin in the presence or absence of a reducing agent, and then precipitated in trichloroacetic acid. The resulting precipitates were collected on nitrocellulose membranes and counted for radioactivity. The radioactivity correlated well with the appearance of a heme-containing peptide in the trypsin digested peptide fragments of cytochrome c. This procedure is simpler and faster than the previously reported methods. PMID- 3032019 TI - Determination and metabolism of dithiol-chelating agents: electrolytic and chemical reduction of oxidized dithiols in urine. AB - The presence of oxidized species of the dithiol-chelating agents, meso-2,3 dimercaptosuccinic acid (DMSA) and 2,3-dimercaptopropane-1-sulfonic acid (DMPS), in human urine was determined by chemical and electrolytic reduction methods. Urine from a human given either DMSA or DMPS was treated with electrolysis, dithiothreitol, or sodium tetrahydridoborate (NaBH4). The SH groups were derivatized with monobromobimane for the determination of unaltered dithiols. Total dithiol (unaltered and oxidized) was determined by reduction followed by derivatization with monobromobimane. The bimane derivatives were identified and quantified by HPLC and fluorescence. Although all three reduction methods gave similar results, electrolytic reduction of oxidized DMSA and chemical reduction with NaBH4 of oxidized DMPS are recommended based upon both day to day reproducibility and recovery of standards. After reduction a 4-fold increase in DMSA and a 20-fold increase in DMPS were found in urine by 12 h after an oral dose of DMSA or DMPS. These new methods for the determination of dithiols and their oxidized forms should lead to a better understanding of the metabolic properties of these increasingly important orally effective chelating agents. PMID- 3032020 TI - Fluorometric assay for ADP-ribosylarginine cleavage enzymes. AB - A continuous fluorometric assay for enzyme activities which remove ADP-ribose linked to proteins at arginine was developed. The substrate analog, N alpha dansyl-N omega-(1,N6-etheno-ADP-ribosyl)arginine methyl ester, was used to assay the catalytic activities of dinitrogenase reductase activating glycohydrolase from Rhodospirillum rubrum and nucleotide pyrophosphatase from Crotalus adamanteus. The assay is based on the increase in fluorescent emission by ethenoadenine accompanying the enzyme-catalyzed hydrolysis of the substrate. The assay has been used to detect activities of 10 fmol substrate cleaved per minute. The substrate anomerizes to give a 40:60 equilibrium of alpha:beta ribosylguanidinium anomers, allowing the determination of enzyme stereospecificity. The substrate was used to determine the kinetic parameters and products of the N-glycohydrolase and the pyrophosphatase. PMID- 3032021 TI - Internal countershock produces myocardial damage and lactate production without myocardial ischemia in anesthetized dogs. AB - The global myocardial extraction of lactate was measured in 13 halothane anesthetized dogs to assess the effect of electric countershock applied directly to the heart. Seven animals received two countershocks of 30 delivered joules each, while six animals were not shocked but were atrially paced to a rate of 190 200, both with and without occlusion of the vena cava to produce a mean arterial pressure of 40-50 mmHg. All animals had substantially positive lactate extraction in the baseline state (36 +/- 10% for countershock group vs. 41 +/- 3% for pacing group). Myocardial lactate extraction reached a markedly negative nadir 2.5 min after countershock (-19 +/- 15%), but returned toward normal by 6 min (10 +/- 6%). Lactate extraction was not significantly changed from baseline in the pacing group. The relationship between changes in regional myocardial blood flow (radiolabeled microspheres) and post-countershock myocardial damage (technetium pyrophosphate uptake) was assessed in six dogs shocked as above. Mean myocardial blood flow was increased minimally immediately after countershock (0.78 +/- 0.08 ml X min-1 X g-1 vs. 1.16 +/- 0.3), but there was no difference in blood flow between damaged and undamaged tissue at either time point. The epicardial-to endocardial ratio of blood flow was unchanged after countershock (0.97 +/- 0.05 vs. 0.99 +/- 0.08). There was no relationship between myocardial damage and either the absolute amount of blood flow after countershock (r = -0.03) or the change in blood flow compared with the pre-shock period (r = 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3032022 TI - Following up. PMID- 3032023 TI - Shedding of enteric coronavirus in adult cattle. AB - Electron microscopic examination of fecal specimens from adult dairy cows indicated seasonal coronavirus shedding. Fifty-two cows from a 300 cow herd were monitored for shedding of coronavirus. Approximately 50% to 60% of the cows monitored shed coronavirus during the winter months (November to March) of the first year of the study. During 2 subsequent years of monitoring the same cows, 20% to 30% of the cows shed coronavirus during the winter months. Virus shedding was not detected during the summer months (July to September). Half of the cows monitored were vaccinated with a modified-live rotavirus-coronavirus-Escherichia coli combination vaccine; however, vaccination did not influence seasonal shedding of coronavirus, as compared with shedding in the nonvaccinated cows. In nonvaccinated cows that calved in the winter months, the incidence of coronavirus shedding increased from 20% to 30% during the last 2 months of gestation to 65% to 70% at parturition. In vaccinated cows, the incidence of shedding did not increase at parturition. PMID- 3032024 TI - Uptake of channel catfish virus from water by channel catfish and bluegills. AB - Radiolabeled channel catfish virus entered the gills of juvenile channel catfish (Ictalurus punctatus) and was then concentrated in the gut and the liver over 48 hours. Diminution of radioactivity was not detected in these tissues over the course of the experiment. Bluegills (Lepomis macrochirus) were capable of clearing the virus during a period of 48 hours. [3H]Thymidine alone had a different distribution in the channel catfish than did labeled virus. PMID- 3032025 TI - Effect of nonadrenal illness on adrenal function in the cat. AB - Adrenal function was assessed by a combined dexamethasone suppression-ACTH stimulation test in 18 healthy cats, 17 diabetic cats, and 19 sick nondiabetic cats. In all groups, plasma cortisol concentrations decreased after dexamethasone was administered and increased after ACTH was administered. There were no significant (P greater than 0.05) differences among groups in time trend changes in cortisol concentration. There was considerable variation in adrenal response between cats in each group. Diabetic cats had more variation in base-line and postdexamethasone plasma cortisol concentrations (P less than 0.05) than did other groups. In sick, nondiabetic cats, cortisol concentrations tended to be higher in cats with hyperthyroidism (P = 0.06) than in cats with other diseases. PMID- 3032027 TI - Swine antibody-dependent cellular cytotoxicity against pseudorabies virus infected cells. AB - In pseudorabies virus (PRV) infection of pigs, antibody-dependent cellular cytotoxicity (ADCC) may be an early defense mechanism. Peripheral blood leukocytes (PBL) and pulmonary macrophages mediate ADCC activity. Antibody dependent cellular cytotoxicity against PRV-infected target cells was assessed, and the effect of infection of cells having an ADCC-effector function was determined. Although pulmonary lavage cells (PLC) had ADCC activity, in vitro infection of PLC led to PRV replication, loss of cell viability, and loss of ADCC activity. In contrast, infection of PBL did not lead to replication, decreased cell viability, or reduced ADCC activity, compared with those in non-infected controls. Measuring ADCC activity in a longitudinal study revealed that PBL from neonates had lower ADCC activity than did PBL from pigs greater than 3.5 months old. Peripheral blood leukocytes and not PLC may have a greater role in control of PRV dissemination in the pig. The difference in activity between cells from neonates and older pigs might explain, in part, the age dependency in the severity of the disease. PMID- 3032026 TI - Determination of thyroxine, triiodothyronine, and cortisol changes during simultaneous adrenal and thyroid function tests in healthy dogs. AB - Changes in thyroxine (T4), triiodothyronine (T3), and cortisol during a combined adrenal (dexamethasone suppression/adrenocorticotrophic hormone response test) and thyroid function tests (thyroid-stimulating hormone [TSH] response test) were determined in 20 healthy hospitalized pet dogs. The effect of dexamethasone on T4 and T3 changes was evaluated during a simultaneous TSH response/dexamethasone suppression adrenocorticotrophic hormone response test. Greater ranges in basal cortisol concentrations and slower changes after dexamethasone was administered were observed in healthy pet dogs kenneled in a hospital setting than those reported for conditioned laboratory dogs. Pet dogs were observed to demonstrate cortisol suppression more reliably at 4 hours than at 2 hours after dexamethasone was administered. Dexamethasone had no effect on the response to TSH as assessed by T4 and T3 assays, thus supporting the validity of combining adrenal and thyroid response tests in a 5-hour period. PMID- 3032028 TI - Experimentally induced ovine border disease: extensive hypomyelination with minimal viral antigen in neonatal spinal cord. AB - Border disease (BD) was experimentally induced in 9 lambs by inoculation of their dams with the BD-31 strain of border disease virus (BDV) at 50 days of gestation. These ewes developed serum-neutralizing antibody titers to BDV. The diagnosis of BD in their lambs was confirmed by hairy birthcoats, intrauterine growth retardation, tremors, and hypomyelination. Three clinically healthy age-matched control lambs, whose dams had been given an inoculum containing only BDV-free cell culture supernatant, were studied in parallel. There were significant differences in birth weights and in the lengths of the right tibiae and radii between the affected and the control lambs. There was a gradient in severity of clinical neurologic signs among the affected lambs, which directly correlated with the severity of hypomyelination in their spinal cords. However, the difference in severity of the neurologic deficits did not correlate with differences either in the precolostral serum-neutralizing antibody titers to BDV in these lambs or in the number of BDV antigen-containing cells in their spinal cords. Only approximately 0.01% to 0.3% of spinal cord cells, both in gray matter and white matter, were BDV antigen positive in the affected lambs. These results indicate that extensive infection of CNS cells with their subsequent destruction or functional alteration may not be a critical part of the pathogenesis of the hypomyelination in BD. PMID- 3032029 TI - Ultrastructural and microprobe analysis of simian lung mite pigments. AB - Using light microscopy, polarizable amorphorous bodies with tinctorial characteristics of lipochromes and flocculent pigmental materials were found within the alveolar macrophages and peribronchiolar region of the lungs in a pig tailed macaque (Macaca nemistrina) infected with simian lung mite (Pneumonyssus simicola). Examination of these pigmental bodies, using a high-voltage (1.2 meV) electron microscope and an energy-dispersive X-ray analysis system, indicated that the pigmental bodies contained a high concentration of silica. One female adult P simicola also was examined, and its digestive tract had visible, polarizable pigment and a high concentration of silica. Thus, lesions associated with lung mite infection in Old World monkeys may be superimposed by this silicotic condition, which may be associated with their arid, dusty, environment. Therefore, until the link between lung mite infection and silicosis is clarified, experimental inhalation toxicologic findings in mite-infected Old World monkeys should be interpreted cautiously. PMID- 3032030 TI - Apparent effect of catalase on airway edema in guinea pigs. Role of endotoxin contamination. AB - The airway edema that develops in guinea pigs after exposure to toluene diisocyanate (TDI) requires the presence of polymorphonuclear leukocytes (PMN). To determine whether this airway edema is mediated by the release of hydrogen peroxide from PMN, we treated animals intravenously with catalase bound to polyethylene glycol and examined the extravasation of Evans blue dye into the tracheal wall after exposure to air or 3 ppm TDI for 1 h. Catalase (25,000, 100,000, and 300,000 IU/kg) caused a dose-dependent inhibition of the TDI-induced increase in dye extravasation. However, treatment with catalase, inactivated at the peroxide binding site with 3-aminotriazole, inhibited dye extravasation after exposure to TDI as effectively as the equimolar 100,000 IU/kg dose of active catalase. Injection of polyethylene glycol alone was without effect. Dose dependent decreases in extravascular migration of PMN and in circulating PMN also were noted after catalase treatment. These results suggest that the catalase preparations used in these studies inhibited the PMN-dependent airway edema by an effect other than hydrogen peroxide scavenging. Examination of this and other commercially available catalase preparations revealed trace concentrations of endotoxin at levels that could be responsible for the observed effects on PMN function. Treatment of animals with doses of Escherichia coli endotoxin similar to those inadvertantly administered to the catalase-treated groups (0.1 ng/kg to 100 ng/kg, intravenously) inhibited TDI-induced extravasation of Evans blue dye in a dose-dependent manner. These results suggest that contaminating endotoxin may contribute to some of the protective effects of preparations of catalase observed in previous studies of vascular permeability. PMID- 3032032 TI - [Poland syndrome and leukemia]. PMID- 3032031 TI - Transpulmonary angiotensin II formation in patients with chronic stable cor pulmonale. AB - The activity of the renin-angiotensin (RA) system and the ability of the lungs to generate angiotensin II (AII) were studied in 11 patients with stable cor pulmonale and respiratory failure caused by chronic obstructive bronchitis and emphysema. Angiotensin I concentrations (18.7 +/- 8.3 pmol/L) were normal, and transpulmonary AII formation rates (TRAIIFR) (14.2 +/- 18.1 pmol/min) were not significantly different from those recorded in nonedematous cardiac subjects (19.9 +/- 20.1 pmol/min), matched for sex, age, and diuretic therapy. The main determinant of TPAIIFR was the mixed venous AI concentration. Administration of oxygen for 30 min led to a small increase in TPAIIFR in the majority of patients. This increase could not be accounted for by changes in mixed venous AI. There was no correlation between serum angiotensin-converting enzyme levels and either the TPAIIFR or the systemic arterial AII concentrations. PMID- 3032033 TI - Chemotherapy alone or chemotherapy with chest radiation therapy in limited stage small cell lung cancer. A prospective, randomized trial. AB - STUDY OBJECTIVE: To determine the effect of concurrent chest radiation therapy on response rate, recurrence, and treatment toxicity among patients with limited stage small cell lung cancer who are receiving combination chemotherapy. DESIGN: Randomized trial with a median follow-up of 57 months. SETTING: A single government institution--the National Cancer Institute. PATIENTS: Consecutive sample of 96 patients with histologically confirmed small cell lung cancer that was confined to the hemithorax of origin or mediastinal and supraclavicular nodes, and which could be encompassed within a tolerable radiation portal ("limited stage"). All patients were followed until death or the end of the study period. INTERVENTIONS: Chemotherapy: Cyclophosphamide, methotrexate, and lomustine in 6-week cycles alternating with vincristine, adriamycin, and procarbazine in 6-week cycles, for a total of 48 weeks. Radiation therapy: Chest irradiation to 40 Gy in 15 fractions over 3 weeks, given simultaneously with the first chemotherapy cycle. MEASUREMENTS AND MAIN RESULTS: The combined therapy led to a significantly higher response rate (complete responses, 81%, compared with partial responses, 43%; 95% Cl for the difference, 20% to 56%), significantly improved local control of the chest tumor (p less than 0.001), and significantly longer survival (p less than 0.035) (median, 15.0 months, compared with 11.6 for chemotherapy alone). The combined therapy produced significantly more myelosuppression, weight loss, esophagitis, and pulmonary dysfunction. There were more infections and deaths from toxicity in the combined treatment group, but the differences between groups were not statistically significant. CONCLUSION: A regimen of combined chemotherapy and chest radiation therapy given concurrently is superior to chemotherapy given alone in inducing remission and prolonging survival in patients with limited stage small cell lung cancer, and the benefit of combined therapy is reduced by its greater toxicity. PMID- 3032034 TI - Long-term adjuvant therapy with tamoxifen in breast cancer: how long is long? PMID- 3032035 TI - Poorly differentiated carcinoma or extragonadal germ cell cancer. PMID- 3032036 TI - Chronic Epstein-Barr virus infection and human immunodeficiency virus infection. PMID- 3032037 TI - Cryptococcal meningitis and fluconazole. PMID- 3032038 TI - Treatment of postmenopausal osteoporosis. PMID- 3032039 TI - [Neuropathies and monoclonal dysglobulinemias]. AB - Nine patients with peripheral neuropathy and monoclonal gammapathy are presented (4 multiple myeloma, 5 Waldenstrom's macroglobulinemia). Two patients had tremor and ataxia. All patients had delayed nerve conduction and increased cerebrospinal fluid protein. Symptoms of neuropathy preceded detection of serum protein abnormalities in seven cases. Nerve fiber lesions involved myelin and axons. Biphasic myelopathy with uniform separation of myelin lamellae was observed in one case of Waldenstrom's disease. Amyloid stains were negative. Treatment was successful in one of four patients. 185 cases of peripheral neuropathy with monoclonal gammapathy are reviewed. Pathogenic role of microvascular changes, amyloid depositions, antimyelin antibody is discussed. The role of anti MAG antibody remains unresolved. Nerve damage due to another cause is possible. The paraprotein and the neuropathy may not be directly related but both caused by the underlying condition. PMID- 3032040 TI - Modeling complex molecular interactions involving proteins and DNA. AB - We have presented a perspective of progress in three areas of simulations of complex molecules: the development of force fields for molecular simulation; the application of computer graphics, molecular mechanics and molecular dynamics in simulations of DNA and DNA-drug complexes and the application of computer graphics, molecular mechanics and quantum mechanics in studies of enzyme substrate interactions. It is our perspective that improvements are being made in force fields, and these will allow a more accurate simulation of structures and energies of complex molecules. In the area of DNA molecular mechanics and dynamics, it is clear that the use of computer graphics model building combined with NMR NOE data is a potentially very powerful tool in accurately determining structures of drug-DNA complexes using molecular mechanics and dynamics. Finally, we are in a position to reasonably simulate structures and (qualitatively) energies for complete reaction pathways of enzymes using a combination of computer graphics, molecular mechanics and quantum mechanics. More accurate energies and pathways are sure to follow, using the combined molecular mechanics/quantum mechanics optimization developed by Singh and the free energy perturbation methods pioneered in Groningen and Houston. PMID- 3032041 TI - The structure of membrane bound cytochrome c oxidase. PMID- 3032042 TI - Interaction of thrombin with thrombomodulin. PMID- 3032043 TI - Complex interaction between factor Va-light chain and thrombomodulin in the regulation of protein C activation. PMID- 3032044 TI - Thrombin-cellular interactions. AB - It is clear that there are a number of different types of reactions between thrombin and the cell surface (TABLE 6). In one type, thrombin binds to cell surface receptors resulting in cellular activation. In other types of reactions, there are at least two components to the thrombin-specific pathway of cellular activation: a classical receptor to which thrombin binds, and a protein that is cleaved. In both types of reactions, thrombin binding and/or proteolysis is linked to changes in GTP-binding proteins, protein kinase C, or other pathways. In most cases, the receptor and membrane substrates involved in cellular activation are not well characterized. In another type of reaction, the interaction between thrombin and proteins in the extracellular fluid is regulated by cell-surface receptors. Binding of thrombin to these receptors can result in acceleration or inhibition of the reactions with the soluble proteins. In the fourth type of reaction, thrombin cleaves a cell-membrane protein that is involved in reactions with plasma proteins. Recognition of the different types of interactions between thrombin and the cell surface is necessary for the correct interpretation of experimental observations. Although the term receptor has classically referred to a cell-surface component to which an agonist binds, it is now clear that there are additional membrane components that specifically bind potential agonists not leading to cellular activation. PMID- 3032045 TI - Thrombin interactions with cultured fibroblasts: relationship to mitogenic stimulation. PMID- 3032046 TI - Thrombin-receptor occupancy initiates a transient increase in cAMP levels in mitogenically responsive hamster (NIL) fibroblasts. AB - Previous studies from our laboratory have shown that thrombin mitogenesis requires both high-affinity receptor occupancy and enzymic activity. Combined addition of DIP-inactivated-thrombin, which retains the ability to bind to thrombin receptors, and enzymically active gamma-thrombin generates a complete set of signals sufficient to initiate cell proliferation. Several possible signals, including stimulation of ion fluxes and phosphoinositide turnover, appear to be stimulated by thrombin's enzymic activity, but not by receptor occupancy. We now report that alpha-thrombin and DIP-thrombin stimulate an early, transient increase of 60 to 200% in intracellular levels of cAMP. This stimulation occurs at low mitogenic concentrations of alpha-thrombin where less than half the receptors are occupied. Enzymically active gamma-thrombin, which stimulates other types of signals, has no stimulatory effects on cAMP. Thus, this effect appears to be generated by high-affinity interaction of thrombin with its cell-surface receptors. Artificially increasing cAMP levels within these cells, however, cannot replace the requirement for thrombin-receptor occupancy in completing the mitogenic stimulation. Therefore, thrombin-receptor occupancy may generate additional, as yet unidentified, required signals. PMID- 3032047 TI - Thrombin-induced alterations in endothelial permeability. AB - Figure 15 summarizes the current understanding of mechanisms of endothelial permeability alterations induced with thrombin. If thrombin generation exceeds the antiprotease activity, thrombin results in clotting of fibrinogen and intravascular fibrin accumulation. Pulmonary neutrophil sequestration also occurs after fibrin deposition, and this is related to the degree and duration of fibrin sequestration. Neutrophil activation appears to be an essential requirement for the mediation of the pulmonary vascular injury. Thrombin-induced intravascular coagulation results in the generation of lipid mediators (LTB4 and HETEs), which may be involved in increasing endothelial permeability. The release of thrombin in higher concentrations during lysis of fibrin (sequence; see text) FIGURE 15. Hypothesis showing mechanisms of thrombin-induced increase in endothelial permeability to proteins. Thrombin may have direct effects on endothelial permeability, or thrombin induced fibrinogen clotting, activation of neutrophils, and the release of lipid metabolites that subsequently lead to an increase in endothelial permeability. clots may induce a direct formation of interendothelial "gaps." Therefore, the vascular injury induced by neutrophil activation and the formation of endothelial "gaps" induced directly by thrombin can both increase the endothelial permeability to proteins. Thrombin is an important mediator of increased endothelial permeability to macromolecules, and may participate in the inflammatory response. In this regard, thrombin may be similar to other mediators (such as histamine and serotonin) that have been previously documented to increase macromolecule transport across the endothelium. The implications of free thrombin in increasing endothelial permeability may be greater because thrombin not only has a direct effect on endothelial permeability, but also induces clotting of fibrinogen and the subsequent generation of mediators that activate neutrophils and that in turn can induce endothelial injury. PMID- 3032048 TI - Thrombin effects on cultured nerve cells: clinical implications and evidence for a novel mechanism of neuronal activation. PMID- 3032049 TI - Hormone-like activity of human thrombin. AB - Recently, we have shown that thrombin is a chemotaxin and growth-promoting agent for cells of the mononuclear phagocytic lineage. These activities are independent of thrombin's enzymatic activity. Unlike other chemotactic factors, thrombin is specific for monocytes and does not attract granulocytes. To further explore the cellular specificity we have used a human leukemia cell line HL-60 that is capable of in vitro differentiation toward either monocytes (HL-60/mono) following incubation with 1,25(OH)2D3, or granulocytes (HL-60/gran) following incubation with DMSO. In contrast to undifferentiated HL-60 cells or HL-60/gran, we find that HL-60/mono respond chemotactically to intact human alpha-thrombin, esterolytically inactive iPR2P-alpha-thrombin, and the thrombin-derived peptide CB67-129, previously shown to contain the thrombin chemotactic exosite. In addition, thrombin induces in HL-60/mono association of actin with the cytoskeleton and causes an increase in levels of free cytosolic Ca2+. These phenomena are well characterized as early events occurring concomitant with directed cell movement associated with exposure to chemotactic agents such as FMLP. Furthermore, in contrast to fibroblasts, both iPR2P-alpha-thrombin and the thrombin chemotactic peptide CB67-129 evoke dose-dependent [3H]TdR incorporation, protein synthesis, and cell replication in growth-arrested J-744 cells, a murine macrophage-like cell line. Limited tryptic digests of CB67-129 lose chemotactic activity but retain full mitogenic activity, demonstrating that as with PDGF, the sites on CB67-129 required for chemotaxis and mitogenesis are clearly dissociable. The mitogenic effects of the CB67-129 digest can be mimicked by a synthetic tetradecapeptide analogue of CB67-129 (residues 367-380) that includes the loop B insertion sequence, previously shown to be critical for thrombin's chemotactic effects. From these data, it is apparent that the loop B insertion is critical for thrombin's nonenzymic biological effects on cells, but additional sites are required for stimulation of cell movement. PMID- 3032050 TI - Thrombin interactions with platelet membrane proteins. PMID- 3032051 TI - Platelet activation by alpha-thrombin is a receptor-mediated event. AB - Computer-assisted data analysis of binding isotherms (LIGAND) has shown that human platelets have binding sites for alpha-thrombin of high (Kd 0.3 nM), moderate (Kd 10 nM), and low affinities (Kd 3 microM). Application of similar techniques has shown that TLCK-thrombin does not, whereas PPACK-thrombin does, bind to the high-affinity binding site accessible to alpha-thrombin, but that both bind to the moderate and low-affinity sites. Treatment of platelets with Serratia marcescens protease destroys the high-affinity site but does not affect moderate-affinity binding. In accordance with this model, both modified thrombins compete with alpha-thrombin for platelet activation at the moderate-affinity site, but only PPACK-thrombin competes at the high-affinity site. These results establish that platelet activation by either low or moderate concentrations of thrombin are receptor-mediated events and explain the paradox of the differential effects of TLCK-thrombin on the binding and activation of platelets by alpha thrombin. PMID- 3032052 TI - Thrombin: active-site topography. PMID- 3032053 TI - Nerve growth factor binding to cells derived from neurofibromas. PMID- 3032054 TI - The nerve growth factor receptor in normal and transformed neural crest cells. PMID- 3032055 TI - Influences of peripheral nerve components on axonal growth. PMID- 3032056 TI - Schwann-like cells cultured from human dermal neurofibromas. Immunohistological identification and response to Schwann cell mitogens. AB - Primary cultures prepared from dermal and plexiform neurofibromas contain Schwann like cells and fibroblast-like cells. SLC are elongated and bipolar or multipolar. By indirect immunofluorescence light microscopy, living SLC bind antibodies against laminin and against nerve growth factor receptor to their surface, but not antibodies against fibronectin. In these respects, cultured SLC are indistinguishable from cultured human adult Schwann cells. FLC are flat and pleomorphic. By indirect immunofluorescence light microscopy, living FLC bind antibodies against fibronectin but not against laminin or NGFR. In these respects, cultured FLC are indistinguishable from cultured human adult endoneurial fibroblasts. Considerable purification of viable SLC from SLC/FLC mixed cultures can be achieved by flow cytofluorometry using a monoclonal anti NGFR antibody. Tritiated thymidine radioautography indicated that mitosis of SLC in mixed SLC/FLC cultures prepared from dermal neurofibromas is infrequent in MEM with 10% calf serum, more frequent in RPMI 1640 medium with 15% fetal calf serum. Central nervous system axolemmal fragments (rat or human) elicited a greater than 10-fold SLC proliferative response in mixed SLC/FLC cultures from three of seven dermal neurofibromas (from six patients with neurofibromatosis), but had no effect on SLC mitosis in cultures from the other four dermal neurofibromas. SLC mitosis was inhibited by concentrations of cyclic adenosine 3',5'-monophosphate analogues known to stimulate proliferation of normal rat Schwann cells. Glial growth factor partially purified from bovine pituitaries stimulated SLC mitosis both in SLC/FLC mixed cultures and in cultures of purified SLC. The studies we have described indicate that neurofibroma SLC can be cultured, unequivocally identified in culture by morphological and immunohistological criteria, purified, and stimulated to proliferate by several Schwann cell mitogens. Further quantitative comparisons of the baseline and mitogen-stimulated rates of proliferation of SLC and age-matched control human Schwann cells are needed, however, to determine which of the two alternate pathogenetic mechanisms for formation of neurofibromas mentioned in the introduction is correct. PMID- 3032058 TI - Enhanced sensitivity of skin fibroblasts from neurofibromatosis patients to transformation by the Kirsten murine sarcoma virus. A potential laboratory assay for individuals at risk of cancer. AB - The present work describes a laboratory assay for individuals predisposed to cancer within NF pedigrees. The assay is based on the association between the increased sensitivity of human skin fibroblasts to transformation by the Kirsten murine sarcoma virus and predisposition to cancer in clinically affected patients and in otherwise apparently healthy individuals within NF pedigrees. The more sensitive the cells are to transformation by KiMSV, the greater the probability that a person from whom such cells have been derived will develop cancer. The results show a strong correlation with the NF trait. Together with the clinical data this laboratory assay could, therefore, be used to ascertain the NF genotype. PMID- 3032060 TI - Hypothalamic-pituitary-adrenal axis and suicide. PMID- 3032059 TI - Molecular and immunocytochemical studies of neurofibromas and related cell types. PMID- 3032057 TI - DNA transfection analysis of the tumors of neurofibromatosis. PMID- 3032061 TI - Isolation and characterization of papillomavirus DNA from nasal inverting (schneiderian) papillomas. AB - To elucidate the possible role of papillomaviruses as etiological agents in nasal inverting papillomas, DNA hybridization techniques were used. Total DNA from two nasal inverting papillomas was examined for the presence of DNA from human papillomavirus (HPV) types 6 and 11 under stringent conditions of hybridization. Both lesions contained DNA hybridizing with HPV 11. Restriction enzyme digestion and subsequent Southern blotting of the DNA samples revealed that one lesion contained viral DNA identical to HPV 11a. The DNA of the other lesion contained an extra 500 base pair insertion. These results provide definitive evidence for the first time for the association of HPV with nasal inverting papillomas. PMID- 3032062 TI - [Cytological study of the mode of contraceptive action of nonoxynol-9]. PMID- 3032063 TI - Long-term potentiation. PMID- 3032064 TI - Perspectives on the discovery of central monoaminergic neurotransmission. PMID- 3032065 TI - The organization and function of the vomeronasal system. PMID- 3032067 TI - Recessive cancer genes and chromosomal mechanisms in tumorigenesis. PMID- 3032066 TI - Genetic instabilities at the chromosomal and the molecular levels induced by the plt oncogene of polyoma virus. PMID- 3032068 TI - 99mTechnetium(V) dimercaptosuccinic acid (99mTc(v) DMSA) as a tumour seeking agent in nasopharyngeal carcinoma. AB - We prepared 99mTechnetium(V) DMSA (pentavalent form) as an imaging agent for eighteen patients with proven nasopharyngeal carcinoma. The head, neck, chest and abdomen were scanned. The nasopharyngeal tumour showed tracer accumulation in only 5 out of the 18 patients (28%). The study indicates that 99mTechnetium(V) DMSA is not a useful radiopharmaceutical for visualising nasopharyngeal tumours although it may have other useful properties. PMID- 3032069 TI - Effect of intrahepatic arterial infusion of 131I-labelled lipiodol on hepatocellular carcinoma of rat. AB - Intrahepatic arterial infusion of 131I-labelled lipiodol was performed to study the intrahepatic distribution of lipiodol and to determine the radiation effect on 3'-Methyl-4-Dimethylaminobenzene (DAB) induced hepatocellular carcinoma (HCC) in rats. From the findings of the softex films and scintigrams of the liver, according to the time course, lipiodol was found to accumulate in the tumour tissues parallel with the degree of perfusion, and it remained in the tumour tissues for an extended period probably due to the delay of the degradation of lipiodol compared to that in non tumour tissues. It was noticed that the lipiodol, accumulated and deposited selectively in the tumour tissues, existed mostly in the extracellular space but not in the intracellular space. In histological studies of the resected specimens of the tumours, complete necrosis of hepatocellular carcinoma was recognized 2 to 8 weeks after the infusion of 131I-labelled lipiodol, while there was none in the control group where cold lipiodol was employed. These results suggest that the most effect on hepatocellular carcinoma in this study is caused by the radiation effect of 131I labelled lipiodol. PMID- 3032070 TI - Biogenic amines as regulators of the proliferative activity of normal and neoplastic intestinal epithelial cells (review). AB - The role of extracellular amines such as noradrenaline and serotonin and their interaction with cyclic nucleotides and intracellular polyamines in the regulation of intestinal epithelial cell proliferation is reviewed with particular reference to the differences between normal and neoplastic cells. In respect to the normal epithelium of the small intestine there is a strong case to support the notion that cell proliferation is controlled by, amongst other things, sympathetic nerves. In colonic carcinomas, antagonists for certain serotonin receptors, for histamine H2 receptors and for dopamine D2 receptors inhibit both cell division and tumour growth. Because of the reproducible variations between tumour lines in the response to these antagonists, this inhibition appears to be due to a direct effect on the tumour cells rather than an indirect effect via the tumour host or stroma. This conclusion is supported by the cytocidal effects of toxic congeners of serotonin on the tumour cells. The most salient difference between the amine responses of normal and neoplastic cells relates to the issue of amine uptake. Proliferation of crypt cells is promoted by amine uptake inhibitors, presumably because they block amine re uptake by the amine secreting cells--sympathetic neurones and enteroendocrine cells. However, tumour cell proliferation is strongly inhibited by amine uptake inhibitors, suggesting that neoplastic cells can, and need to take up the amine before being stimulated by it. Recent revelations in the field of oncogenes also support an important association between amines, cyclic nucleotides and cell division. The ras oncogenes code for a protein that is a member of a family of molecules which relay information from extracellular regulators, such as biogenic amines, to the intracellular regulators, including cyclic nucleotides. Evidence is presented suggesting that enteroendocrine cells, enterocytes, carcinoid tumour cells and adenocarcinoma cells all have the same embryonic origin and that cells exhibiting an admixture of endocrine and proliferative properties exist in colonic tumours, but not in the normal intestinal epithelium. Thus, it appears that in the normal intestine a clear structural and functional distinction exists between the regulating cells (i.e. the sympathetic neurones and enteroendocrine cells) and the regulated cells (i.e. the undifferentiated crypt cells): cells that have acquired a regulating role are no longer able to divide and cells which are able to divide do not take up or store amines.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3032071 TI - The influence of progestins on receptor levels in breast cancer metastasis. AB - The influence of oral high-dose (HD) medroxyprogesterone acetate (MPA) and megestrol acetate (MA) treatment on steroid hormone receptor levels in metastatic breast tumor tissue was investigated. In ten postmenopausal women with advanced breast cancer, receptor biopsies were obtained from the same tumor before the start, and one and eight weeks after the start of oral HD progestin treatment. Endocrine treatment had been stopped in all patients at least five weeks before the start of progestin treatment. Estradiol receptor (ER) levels were reduced by 26.9% and 20.0% respectively after one and eight weeks of treatment. Progesterone receptors (PgR) were significantly reduced to undetectable levels, possibly due to MPA and MA binding to PgR. A stepwise decrease in androgen receptors (AR) was observed, indicating that the two progestins (MPA and MA) may also act through AR, and/or interfere with the replenishment of AR. PMID- 3032072 TI - Flow cytometric analysis of primary lung carcinomas and their lymph node metastases. AB - Specimens of primary lung carcinomas and lymph node metastases from the same 18 patients were investigated by means of flow cytometry. The number of DNA stemlines, DNA indices, the proportion of diploid cells in the tumors and the distribution of the cell cycle phases were compared. In 10 patients DNA stemlines and DNA indices were identical in primary tumors and metastases. In two cases the DNA indices were doubled in metastases. In 6 cases the primary tumors contained two abnormal DNA stemlines and their metastases contained only one aneuploid stemline. Gross differences between primary tumors and lymph node metastases with regard to the proportion of cell cycle phases could not be found. The large variation between primary tumors and lymph node metastases with regard to DNA stemlines indicates that flow cytometric analysis of lymph nodes gives only limited information about the primary tumors. PMID- 3032073 TI - Differential effects of phorbol ester on tumor cells induced by avian sarcoma virus. AB - Avian sarcoma virus-induced tumors usually grow progressively for several weeks and then regress. We have injected phorbol myristate acetate (PMA) directly into tumors in an effort to stimulate neoplastic growth. The results show instead that PMA exerted an inhibitory effect in this regard and, in fact, caused an acceleration of tumor regression. At the same time, treatment of cultured avian sarcoma cells with PMA resulted in greatly diminished levels of the kinase activity associated with the src gene product, pp60src. PMA-treated tumor cells from regressing sarcomas were, however, stimulated to express viral antigens at their surface and produced more progeny virus than did untreated tumor cells. PMID- 3032074 TI - Calcium as a mediator of tubuloglomerular feedback. PMID- 3032075 TI - Role of H+ and HCO3- in salt transport in gallbladder epithelium. PMID- 3032077 TI - Peripheral airway ganglia. PMID- 3032076 TI - Proton and bicarbonate transport mechanisms in the intestine. PMID- 3032078 TI - Nervous receptors of the tracheobronchial tree. PMID- 3032079 TI - Spectroscopic probes of muscle cross-bridge rotation. PMID- 3032080 TI - The nature and identity of the internal excitational transmitter of vertebrate phototransduction. PMID- 3032081 TI - The molecular mechanism of visual excitation and its relation to the structure and composition of the rod outer segment. PMID- 3032082 TI - Molecular properties of the cGMP cascade of vertebrate photoreceptors. PMID- 3032083 TI - Role of proton and bicarbonate transport in pancreatic cell function. PMID- 3032084 TI - [Structural-functional research on polymyxins. 1H-NMR spectra of polymyxin B and its shortened analog]. AB - Polymyxin B and its shortened analog were studied comparatively by 1H-NMR spectroscopy. Analysis of the signal chemical shifts, constants of spin-spin interaction of 3J HN-C alpha H and temperature coefficients of the NH signal chemical shifts revealed absolute structural identity of both molecules cyclic parts. This proved that there was no conformative interaction between the cyclic and linear parts of the polymyxin B molecule. Comparison of the results with the data on the biological activity showed that the hydrophobic N-end moiety of the polymyxin B molecule played a specific role in its antibacterial effect and toxicity. PMID- 3032085 TI - [Detection of an inhibitor of the causative agent of plague active against strains of the plague microbe from an homologous focus. The effect of lithium chloride on detecting the inhibitor]. AB - A nutrient medium containing 0.7 per cent of lithium chloride was tested and Yersinia pestis inhibitor was detected in 83.3 per cent of the strains on the growth tufts of 8 indicator strains isolated from a homologous focus. Lowering the lithium chloride concentration in the medium up to 0.6 per cent promoted detection of the inhibitor in a larger number of the tested strains (93.3 per cent). But even on this medium none of the indicator strains formed overall growth tufts. The subcultures of the indicator strains resistant to lithium chloride formed overall growth tufts on agar plates with lithium chloride and were sensitive to the inhibitor of the tested strains from the central Caucasus. However, during storage they lost their sensitivity to the inhibitor which did not permit their use as the test cultures. PMID- 3032086 TI - Therapeutic implications of inhibition versus killing of Mycobacterium avium complex by antimicrobial agents. AB - Patients with the acquired immune deficiency syndrome (AIDS) with disseminated Mycobacterium avium infection have responded poorly to treatment with rifabutine (Ansamycin) and clofazimine, in spite of the good in vitro response of M. avium to these antimicrobial agents. We compared the ability of these and other antimicrobial agents to kill versus the ability to inhibit the growth of strains of the M. avium complex isolated from patients with AIDS. Killing curve experiments showed that the concentrations of rifabutine and clofazimine needed to kill two log units of M. avium are at least 32 times greater than the concentrations needed to inhibit growth. Little or no killing occurred at concentrations of these antimicrobial agents that are achievable in serum. In contrast, five of seven strains tested were killed by ciprofloxacin at concentrations that can be achieved in serum. Ciprofloxacin should be studied further for possible use in the treatment of M. avium infections. PMID- 3032087 TI - Role of porins in intrinsic antibiotic resistance of Pseudomonas cepacia. AB - The measured outer membrane permeability of Pseudomonas cepacia to the beta lactam nitrocefin was low: approximately 10 times less than that of Escherichia coli and comparable to that of Pseudomonas aeruginosa. The purified P. cepacia porin demonstrated an average single channel conductance in 1 M KCl of 0.23 nS. PMID- 3032088 TI - Rifabutin and rifapentine compared with rifampin against Mycobacterium leprae in mice. PMID- 3032089 TI - Activity of (+)-cyclaradine (Sch 31172) against herpes simplex virus in vitro and in vivo. AB - (+)-Cyclaradine (Sch 31172) is the carbocyclic derivative of adenosine arabinoside (9-beta-D-arabinofuranosyladenine). Because it is not deaminated by deaminase in serum, as is adenosine arabinoside, (+)-cyclaradine is about 2 to 5 times more active in vitro against herpes simplex virus. (+)-Cyclaradine has in vitro activity nearly equivalent to that of phosphonoformate but is significantly less active than acycloguanosine (acyclovir; ACV), trifluorothymidine, or 9-(1,3 dihydroxy-2-propoxymethyl)guanine. The absolute ratios of in vitro activities are difficult to determine because of variability among virus strains, inoculum size, and dependence on the tissue culture cell line in which the comparative test is carried out. (+)-Cyclaradine is active against TK-, ACV-resistant mutants. In the guinea pig model of vaginal herpes simplex virus infection, (+)-cyclaradine is only slightly less active than ACV when both molecules are nearly equivalently bioavailable; thus, the large difference in activity seen in vitro is not reflected in this in vivo model system. PMID- 3032090 TI - Inactivation of enveloped viruses and killing of cells by fatty acids and monoglycerides. AB - Lipids in fresh human milk do not inactivate viruses but become antiviral after storage of the milk for a few days at 4 or 23 degrees C. The appearance of antiviral activity depends on active milk lipases and correlates with the release of free fatty acids in the milk. A number of fatty acids which are normal components of milk lipids were tested against enveloped viruses, i.e., vesicular stomatitis virus, herpes simplex virus, and visna virus, and against a nonenveloped virus, poliovirus. Short-chain and long-chain saturated fatty acids had no or a very small antiviral effect at the highest concentrations tested. Medium-chain saturated and long-chain unsaturated fatty acids, on the other hand, were all highly active against the enveloped viruses, although the fatty acid concentration required for maximum viral inactivation varied by as much as 20 fold. Monoglycerides of these fatty acids were also highly antiviral, in some instances at a concentration 10 times lower than that of the free fatty acids. None of the fatty acids inactivated poliovirus. Antiviral fatty acids were found to affect the viral envelope, causing leakage and at higher concentrations, a complete disintegration of the envelope and the viral particles. They also caused disintegration of the plasma membranes of tissue culture cells resulting in cell lysis and death. The same phenomenon occurred in cell cultures incubated with stored antiviral human milk. The antimicrobial effect of human milk lipids in vitro is therefore most likely caused by disintegration of cellular and viral membranes by fatty acids. Studies are needed to establish whether human milk lipids have an antimicrobial effect in the stomach and intestines of infants and to determine what role, if any, they play in protecting infants against gastrointestinal infections. PMID- 3032091 TI - Penetration of new azole compounds into the eye and efficacy in experimental Candida endophthalmitis. AB - We studied the penetration of three azole compounds, ketoconazole, itraconazole, and fluconazole, into the ocular tissues and fluids of rabbits in the presence and absence of ocular inflammation. Drug concentrations were compared with those found in serum and cerebrospinal fluid. The rank order of penetration into eye tissue was fluconazole greater than ketoconazole greater than itraconazole. Fluconazole penetrated freely into both inflamed and uninflamed eyes. The presence of inflammation improved penetration of all three compounds into ocular fluids and tissues. Penetration of these azoles into the anterior chamber of uninflamed eyes and into the cerebrospinal fluid was similar. All three azole compounds reduced the number of yeasts found in the eye in hematogenous Candida albicans endophthalmitis in rabbits when therapy was initiated within 24 h of inoculation. However, only ketoconazole significantly reduced yeast counts in the eye when therapy was postponed for 7 days. PMID- 3032092 TI - Inhibiting effect of (RS)-9-[4-hydroxy-2-(hydroxymethyl)butyl]guanine on varicella-zoster virus replication in cell culture. AB - The activity and mode of action of the new nucleoside analog (RS)-9-[4-hydroxy-2 (hydroxymethyl)butyl]guanine (2HM-HBG) against varicella-zoster virus (VZV) were determined. In cell culture, replication of different strains of VZV was inhibited to 50% by 0.4 to 0.7 microM 2HM-HBG, while 685 microM was required to inhibit 50% of the DNA synthesis in uninfected human lung fibroblasts. A thymidine kinase-negative VZV strain was not inhibited by 100 microM 2HM-HBG. Inhibition of VZV replication was not reversible after 7 to 14 days of incubation, depending on the multiplicity of VZV. 2HM-HBG was shown to be selectively phosphorylated by purified VZV thymidine kinase, with an inhibition constant of 32.5 microM. The antiviral activity of 2HM-HBG in cell culture was decreased by the addition of deoxythymidine and deoxycytidine but not by other ribo- or deoxyribonucleosides. PMID- 3032093 TI - Decreased binding of antibiotics to lipopolysaccharides from polymyxin-resistant strains of Escherichia coli and Salmonella typhimurium. AB - The interactions of polycationic antibiotics with lipopolysaccharide (LPS) isolated from parental and polymyxin-resistant strains of Salmonella typhimurium and Escherichia coli were measured by using a cationic spin probe. Electron spin resonance spectra indicated that increasing concentrations of cations competitively displaced probe from LPS aggregates. Polymyxin B and other cations displaced less probe from LPS of polymyxin-resistant strains than from LPS of the parental strains, whereas the same amount or more probe was displaced from isolates of the mutants by the structurally similar antibiotic, EM 49 (octapeptin). In general, the differential affinities of these antibiotics for LPS correlated with their antibiotic activity in vivo, suggesting that resistance results from a decrease in antibiotic permeability across the outer membrane due to alterations in the LPS which affect antibiotic binding. The alterations in the structure of LPS from the polymyxin-resistant mutants of E. coli were characterized using 31P nuclear magnetic resonance spectroscopy. The results suggested that esterification of the core-lipid A phosphates is responsible for increased resistance to polymyxin B and that this alteration is different from that previously proposed for the S. typhimurium strains. In both cases, however, resistance was the result of modifications that result in a less acidic lipid A. PMID- 3032095 TI - Organization of two sulfonamide resistance genes on plasmids of gram-negative bacteria. AB - The organization of two widely distributed sulfonamide resistance genes has been studied. The type I gene was linked to other resistance genes, like streptomycin resistance in R100 and trimethoprim resistance in R388 and other recently isolated plasmids from Sri Lanka. In R388, the sulfonamide resistance gene was transcribed from a promoter of its own, but in all other studied plasmids the linked genes were transcribed from a common promoter. This was especially established with a clone derived from plasmid R6-5, in which transposon mutagenesis showed that expression of sulfonamide resistance was completely dependent on the linked streptomycin resistance gene. The type II sulfonamide resistance gene was independently transcribed and found on two kinds of small resistance plasmids and also on large plasmids isolated from clinical material. PMID- 3032094 TI - Molecular genetic analysis of cephalosporinase production and its role in beta lactam resistance in clinical isolates of Enterobacter cloacae. AB - Two strains of Enterobacter cloacae were isolated from a patient before (strain MHN1) and during (strain MHN2) treatment with moxalactam and gentamicin. Strain MHN1 exhibited inducible ampC cephalosporinase production. In contrast, strain MHN2 expressed the enzyme constitutively at a 3,000-fold higher level. With the Escherichia coli ampC gene as a hybridization probe it was shown that the genomic arrangement of the ampC region was the same in both strains. To gain more insight into regulatory phenomena, the ampC genes were cloned, and their expression was studied in E. coli K-12. The ampC gene from MHN1 behaved normally and conferred inducible beta-lactam resistance. A regulatory region of at least 800 base pairs involved in controlling repression-induction was located immediately upstream of ampC. Surprisingly, when present in E. coli the ampC gene from MHN2 no longer overproduced the cephalosporinase, and inducible expression was observed. This indicates that in MHN2 stable cephalosporinase overproduction is controlled by another factor which is not linked to the ampC gene. PMID- 3032096 TI - Acyclic guanosine analogs as inhibitors of human cytomegalovirus. AB - The acyclic guanosine analogs R- and S-enantiomers of 9-(3,4 dihydroxybutyl)guanine [(R)- and (S)-DHBG], 9-(4-hydroxybutyl)guanine (HBG), and 9-(2-hydroxyethoxymethyl)guanine (ACV) were examined for their effects on human cytomegalovirus (CMV) replication and on CMV DNA synthesis in cell culture as well as for their ability as triphosphates to interact with CMV DNA polymerase. Production of early CMV antigens was not affected. All analogs inhibited CMV DNA synthesis and late viral antigen synthesis. Primary CMV isolates were less susceptible to all tested analogs than was the laboratory strain CMV Ad.169. The triphosphate of ACV was the most potent inhibitor of CMV DNA polymerase, with an observed Ki of 0.0076 microM. The corresponding Ki values of the triphosphates of (R)-DHBG, (S)-DHBG, and HBG were 3.5, 13.0 and 0.23 microM, respectively. All triphosphates of the analogs given above inhibited CMV DNA polymerase in a competitive manner with respect to dGTP. The triphosphates of the analogs also inhibited reactions when the synthetic template poly(dC)oligo(dG)12-18 was used, whereas no inhibition was observed with poly(dA)oligo(dT)12-18. None of the triphosphate analogs supported DNA synthesis in the absence of dGTP, showing that no analog was an alternative substrate to dGTP. PMID- 3032097 TI - Activity of cefazolin and two beta-lactamase inhibitors, clavulanic acid and sulbactam, against Bacteroides fragilis. AB - One hundred clinical isolates of the Bacteroides fragilis group of bacteria were tested by agar dilution for susceptibility to cefazolin alone or in combination with clavulanic acid or sulbactam. For cefazolin, the MIC for 50% of the isolates (MIC50) was 32 micrograms/ml, the breakpoint for susceptibility. With the addition of 0.5 micrograms of clavulanic acid per ml, the MIC for 90% of the isolates (MIC90) was 8 micrograms/ml, well within the achievable range of concentrations in serum or tissue. Similarly, with the addition of 0.5 micrograms of sulbactam per ml, the MIC90 was 16 micrograms/ml. The addition of a higher concentration (4.0 micrograms/ml) of clavulanic acid or sulbactam resulted in MIC90S which were fourfold lower than those with 0.5 micrograms/ml. A fixed ratio of cefazolin-beta-lactamase inhibitor of 4:1 resulted in an MIC50 and MIC90 which were intermediate between the 0.5- and 4.0-micrograms/ml fixed concentration of beta-lactamase inhibitor. PMID- 3032098 TI - Inhibition of DNA gyrase by optically active ofloxacin. AB - Inhibition of DNA gyrase activity by optically active ofloxacins was studied and compared with the inhibition of norfloxacin and ciprofloxacin. The (-)-isomer of ofloxacin inhibited the supercoiling activity of gyrase from Micrococcus luteus more effectively than did the (+)-isomer. The 50% inhibitory concentrations of ( )-, (+/-)-, and (+)-ofloxacin; norfloxacin; and ciprofloxacin for gyrase from Escherichia coli were 0.78, 0.98, 7.24, 0.78, and 1.15 microgram/ml, respectively. These values correlated well with the antibacterial activity of each compound against intact bacterial cells. PMID- 3032099 TI - In vitro activity of LY146032, a new lipopeptide antibiotic, against gram positive cocci. AB - The activity of LY146032, a new lipopeptide antibiotic, was compared in vitro with those of vancomycin, oxacillin, and ampicillin against 261 staphylococcal and 154 streptococcal isolates. The MICs of LY146032 and vancomycin were similar, but the bactericidal activity and killing kinetics of LY146032 were more pronounced. PMID- 3032100 TI - Effect of ganciclovir [9-(1,3-dihydroxy-2-propoxymethyl)guanine] on viral DNA and protein synthesis in cells infected with herpes simplex virus. AB - The effect of ganciclovir [9-(1,3-dihydroxy-2-propoxymethyl)guanine] on herpes simplex virus type 1 DNA and protein synthesis was studied. Ganciclovir markedly inhibited the synthesis of viral DNA and gamma proteins in a dose-dependent manner. However, the synthesis of viral beta proteins was significantly increased by ganciclovir in the later stage of infection. PMID- 3032101 TI - Lactobacilli as effectors of host functions: no influence on the activities of enzymes in enterocytes of mice. AB - Preparations of lactobacilli are often used as dietary supplements to improve the growth and efficiency in utilizing food of animals of commercial value. We tested in an experimental model whether the effects of lactobacilli on growth of and food utilization by animals may be due to alteration of the activities of absorptive enzymes in the small bowel. Germfree mice housed in isolators under tightly controlled conditions were monoassociated with one of four strains of indigenous Lactobacillus spp. From 1 to 5 weeks later, the activity of alkaline phosphatase was assayed in homogenates of segments of the upper small intestines of the associated animals and of matched germfree controls. The specific activity of the enzyme was the same in the mice in the two groups. In other experiments, epithelial cells were isolated from the upper small intestines of mice associated with eight Lactobacillus strains (octa-associated) and from those of matched germfree mice and assayed for alkaline phosphatase, phosphodiesterase, and thymidine kinase activities. The epithelial cells were harvested sequentially from the tips of the villi toward the crypts of Lieberkuhn of the intestines. In all preparations, mice of both types yielded an equivalent mass (wet weight) of cells. The protein content of the cells reflected the mass. The activities of the microvillous membrane enzymes alkaline phosphatase and phosphodiesterase and the cytosol enzyme thymidine kinase were the same whether or not the animals contained the bacteria. Therefore, any effects on animal growth and food utilization observed when lactobacilli are used as dietary supplements may not be due to a direct alteration by the bacteria of the absorptive enzymes of the host animal.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3032103 TI - Purification and characterization of deoxyuridine triphosphate nucleotidohydrolase from anemic rat spleen: a trimer composition of the enzyme protein. AB - Deoxyuridine triphosphate nucleotidohydrolase (dUTPase) was purified to near homogeneity from the spleens of rats made anemic by phenylhydrazine injection; the enzyme activity in these spleens was about 30 times higher than that in spleens of untreated rats. The purified enzyme preparation showed an apparent molecular weight of 58,500 and appeared to consist of three identical subunits each with a molecular weight of about 19,500. The purified enzyme catalyzed specifically the hydrolysis of dUTP, and no other naturally occurring nucleoside triphosphates could be hydrolyzed by this enzyme. The Km value for dUTP was 12 microM. Enzyme activity was inhibited by the addition of EDTA, whereas the enzyme preparation exhibited activity in the absence of added divalent cations. Activity was not affected by the addition of fluoride ion. PMID- 3032102 TI - Protease-deficient Bacillus subtilis host strains for production of Staphylococcal protein A. AB - We constructed strains of Bacillus subtilis which produced very low levels of extracellular proteases. These strains carried insertion or deletion mutations in the subtilisin structural gene (apr) which were constructed in vitro by using the cloned gene. The methods used to construct the mutations involved the use of a plasmid vector which allowed the selection of chromosomal integrates and their subsequent excision by homologous recombination to effect replacement of the chromosomal apr gene by a derivative carrying an inactivating insert with a selectable marker (a cat gene conferring chloramphenicol resistance). The strains produced no subtilisin, no detectable extracellular metalloprotease activity, and residual extracellular serine protease levels as low as 0.5% of that of the standard strain from which they were derived. The strains proved to be superior host strains for the production of staphylococcal protein A, accumulating higher levels of intact protein than do previously available B. subtilis strains. PMID- 3032105 TI - Modulation of rat brain cytosolic phosphatidate phosphohydrolase: effect of cationic amphiphilic drugs and divalent cations. AB - The effects of three cationic amphiphilic drugs on rat brain cytosolic phosphatidate phosphohydrolase and their mechanisms of action were studied utilizing membrane-bound, emulsified, and emulsified sonicated phosphatidate as substrates. With the membrane-bound substrate, chlorpromazine, desmethylimipramine, and propranolol inhibited the activity in a dose-dependent fashion with an IC50 of 30-50 microM. In the presence of the emulsified substrate, chlorpromazine was a more potent inhibitor than desmethylimipramine or propranolol but 200 microM was needed for 50% inhibition of activity. Addition of heat-inactivated microsomes to the emulsified substrate, to simulate the conditions with the membrane-bound substrate, did not alter this value. Both Mg2+ and Ca2+ stimulated the enzyme activity but only Ca2+ counteracted the effect of chlorpromazine. Kinetic studies indicate that chlorpromazine acts as a noncompetitive inhibitor of the enzyme. Emulsified sonicated phosphatidate was a good substrate at low (less than 10 microM) concentrations. It was a poor substrate at 1 mM, but at this concentration chlorpromazine stimulated the activity instead of inhibiting. This drug altered the integrity of phosphatidate vesicle membranes as visualized by electron microscopy. The different results obtained with the three types of substrate indicate the importance of the configuration of phosphatidate for the expression of enzyme activity and for its susceptibility to the action of cationic amphiphilic drugs. PMID- 3032104 TI - Early metabolic effects and mechanism of ammonium transport in yeast. AB - Studies were performed to define the effects and mechanism of NH+4 transport in yeast. The following results were obtained. Glucose was a better facilitator than ethanol-H2O2 for ammonium transport; low concentrations of uncouplers or respiratory inhibitors could inhibit the transport with ethanol as the substrate. With glucose, respiratory inhibitors showed only small inhibitory effects, and only high concentrations of azide or trifluoromethoxy carbonylcyanide phenylhydrazone could inhibit ammonium transport. Ammonium in the free state could be concentrated approximately 200-fold by the cells. Also, the addition of ammonium produced stimulation of both respiration and fermentation; an increased rate of H+ extrusion and an alkalinization of the interior of the cell; a decrease of the membrane potential, as monitored by fluorescent cyanine; an immediate decrease of the levels of ATP and an increase of ADP, which may account for the stimulation of both fermentation and respiration; and an increase of the levels of inorganic phosphate. Ammonium was found to inhibit 86Rb+ transport much less than K+. Also, while K+ produced a competitive type of inhibition, that produced by NH4+ was of the noncompetitive type. From the distribution ratio of ammonium and the pH gradient, an electrochemical potential gradient of around 180 mV was calculated. The results indicate that ammonium is transported in yeast by a mechanism similar to that of monovalent alkaline cations, driven by a membrane potential. The immediate metabolic effects of this cation seem to be due to an increased [H+]ATPase, to which its transport is coupled. However, the carriers seem to be different. The transport system studied in this work was that of low affinity. PMID- 3032106 TI - [Analysis of drug sensitivity by successive colony-forming assay]. AB - It is difficult to predict the adequate doses of effective drugs which must be administered in cancer chemotherapy. We have developed a successive colony forming assay in order to analyze changes of drug sensitivity in human cancer cell lines, and it is used as a model of cancer chemotherapy. Presently, widely used methods include the clonog ceni assay as in vitro sensitivity test established by Salmon and Hamberger et al., for revealing effective drugs. However, formation of a second colony using initial colony-forming cells has not yet been performed. We therefore analyzed the changes occurring in drug sensitivity in 2nd and 3rd colony-forming assays. Using a human lung adenocarcinoma cell line PC-9 (established by Hayata at the Surgical Department in Tokyo Medical School), we measured the changes in sensitivity to CDDP and its derivatives. After picking up the colony-forming cells by micropipette and culturing them in culture medium, we tried to incubate them for 1 hour with the anticancer drug, and then replate them in soft agar. This technique is called "2nd colony-forming assay". We were able to carry this out three times. The results obtained showed that the anti-tumor effect of 254S was the same as that of CDDP, whereas that of CBDCA was less than either of them. Although these tendencies were noticeable even in the 2nd colony-forming assay, the sensitivities to CDDP and its derivatives were diminished in the 3rd colony forming assay. PMID- 3032107 TI - [Establishment of adriamycin-resistant human small-cell lung cancer cells in culture: analysis of the mechanism of resistance and cross-resistance]. AB - An adriamycin-resistant cell line was established in culture by continuous exposure of SBC-3 cells, a cell line of human small-cell lung cancer, to increasing concentrations of adriamycin, followed by a cloning procedure. The resistant cells (SBC-3/ADM) were 30 times more resistant to the drug than the parent cells in terms of 70% lethal dose, determined by soft agar clonogenic assay. Uptake studies using [3H] daunomycin, which was completely cross-resistant to adriamycin, showed decreased influx and enhanced efflux of the agent. This resistance to adriamycin may be attributed to an alteration in membrane transport, resulting in reduced intracellular accumulation of the drug. Using the SBC-3/ADM cells, the activity of a variety of drugs was analyzed by soft agar clonogenic assay in order to search for a means of circumventing drug resistance. The SBC-3/ADM cells were markedly resistant to anthracycline antibiotics such as THP-adriamycin and 4'-epi-adriamycin. The cells were also resistant to structurally or pharmacodynamically unrelated compounds such as vincristine, mitomycin C, 40497S, an active compound of ifosfamide, and etoposide. However, the cells were as sensitive to mitoxantrone as the parent cells, and were considerably susceptible to cisplatin. These results suggest that mitoxantrone and cisplatin may exert a sufficient degree of activity for use against small cell lung cancer resistant to adriamycin. PMID- 3032109 TI - [The regression of medial smooth muscle in host arteries supplying human cancers and its functional significance]. AB - The hemodynamics prevailing in cancer tissue are known to be quite abnormal, at least in some cases, in that the tumor vessels are inert to vasoactive stimuli, a feature that has been exploited in the development of pharmacoangiography or induced hypertension chemotherapy. However, not only has this feature yet to be correlated with the structural changes occurring in the vascular wall, but also the remarkable differences existing among cancer patients in its manifestation await future elucidation. In a series of human cancers, we have been aware of peculiar degenerative changes differently involving the medial smooth muscle of tumor-supplying host arteries, which we have assumed act together with the muscular, newly formed "tumor vessels" in creating the abnormal reactivity of cancer blood flow, at the same time accounting for its wide variability. From this viewpoint, cancer-supplying host arteries were submitted to morphometry of their medial smooth muscle in 30 surgical specimens of carcinoma of the stomach or colon, and in 30 autopsied livers with hepatocellular carcinoma. In GI cancers it was disclosed that the percentage volume of smooth muscular cells in the medial layer, as high as 80% in segments of unaffected artery located at a distance from a cancer, rapidly fell to 30% or less as it reached and penetrated into a tumor, and that at the tumor center, a host artery was often transformed into a dilated sac with a purely collagenous wall. Morphometry of hepatocellular carcinoma also demonstrated unambiguous atrophy of the medial muscular layer of host hepatic arteries, which was shown to be significantly thinner than in control specimens even at a distance from the cancer. This too, was interpreted to be a contributory factor in the cancer circulation typically devoid of regulatory activity. We stress that the functional significance of these features of tumor-supplying host arteries should be taken into consideration when studying the behavior of individual cancers in arteriography, pharmacoangiography or induced hypertension chemotherapy. PMID- 3032108 TI - [Serial measurements of serum carcinoembryonic antigen (CEA) and neuron-specific enolase (NSE) during chemotherapy of patients with inoperable lung cancer]. AB - Serial measurements of serum CEA levels were analyzed in 226 patients with inoperable lung cancer (115 small cell carcinomas, 64 adenocarcinomas, 37 squamous cell carcinomas and 10 large cell carcinomas) during chemotherapy. Of all patients, 29.1% had pretreatment CEA levels greater than or equal to 5 ng/ml. In all of the patients with complete response, and 15 (68.2%) of 22 patients with partial response whose pretreatment CEA levels were 5 ng/ml or higher, CEA levels fell to below 5 ng/ml. All of 17 patients who showed a decrease greater than 50% in serum CEA levels during chemotherapy showed a shrinkage of more than 50% in the tumor burden. Serial serum CEA level measurements were useful as an indicator of response to chemotherapy in advanced lung cancer. Serial serum NSE levels were measured in 36 patients with small cell lung cancer. Pretreatment NSE levels were elevated to more than 10 ng/ml in 83.1% of all patients. A transient rise in serum NSE levels occurred in 22 out of 33 patients measured on the third day during initial chemotherapy. Serum NSE levels greater than or equal to 10 ng/ml declined to within the normal range in all patients responding to the chemotherapy. The survival in patients whose NSE levels (greater than or equal to 10 ng/ml) fell to within the normal range for more than four weeks was longer than that in other patients. Serial measurements of serum NSE levels were thus useful for monitoring the response to chemotherapy in cases of small cell lung cancer. PMID- 3032110 TI - [Clinical study on the effect of 24-hour continuous intravenous infusion of CDDP in far-advanced and recurrent carcinoma of the gastrointestinal tract]. AB - This clinical study was undertaken in order to evaluate the effect of CDDP on 43 patients with far-advanced or recurrent carcinoma of the gastrointestinal tract. For all these patients, CDDP at 100 mg/m2 had been administered by continuous intravenous infusion for 24 hours and repeated one to seven times at an interval of 3 to 4 weeks. The effect of this therapy was assessed according to the criteria of clinical evaluation of chemotherapy for solid cancers by Koyama and Saito. The response rate for both complete and partial response was 27.9% among all 43 patients, including 47.1% (8/17) for gastric cancer, 33.3% (1/3) for esophageal cancer, 25.0% (1/4) for hepatocellular carcinoma, 25.0% (1/4) for carcinoma of the gallbladder or bile duct, 20.0% (1/5) for pancreatic cancer and none (0/10) for colorectal cancer. In particular, a good response rate of 37.5% (3/8) was obtained for patients with recurrent tumor and one of 33.3% (6/18) for those with palliative resection of the primary tumor, which was much higher that the rate of 17.6% (3/17) for those without resection. As for the side effects of CDDP therapy, gastrointestinal symptoms were most frequently found in 78.3% of patients followed by bone marrow suppression in 15.2%, and abnormalities of hepatic and renal function in 4.3% and 4.3%, respectively. Consequently, 24-hour continuous intravenous infusion of CDDP was considered to be effective for far advanced or recurrent carcinoma, especially in cases of gastric cancer. PMID- 3032112 TI - [Usefulness of caerulein in suppressing post-TAE complications of the gallbladder]. AB - While transcatheter hepatic arterial embolization (TAE) has been extensively performed as a form of treatment for nonresectable malignant hepatic tumors, complications, such as abdominal pain, fever or leukocytosis due to gallbladder infarction by embolic materials frequently occur and have not yet been overcome. We devised a new procedure for reducing the incidence of gallbladder infarction by administering caerulein prior to TAE. Between 1984 and 1986, 63 patients with hepatocellular carcinoma were treated by TAE with the use of Gelfoam. These patients were divided into 3 groups. Fourteen patients underwent TAE in which the tip of the catheter was placed in the right hepatic artery distal to the origin of the cystic artery (group A). In the other patients the tip of the catheter was placed proximal to the origin of the cystic artery; 40 patients were not treated by caerulein (group B); 9 patients were administered caerulein 20 micrograms intramuscularly 15 to 30 minutes prior to TAE. The incidence of complications after TAE, such as abdominal pain, fever over 38 degrees C, leukocytosis and ultrasonographical abnormalities of the gallbladder was compared in these 3 groups. The results showed that in group C (TAE after administration of caerulein), the incidence of complications was significantly decreased compared with group B(TAE without caerulein). The authors suggest that post-TAE infarction of the gallbladder is effectively diminished by contracting it with caerulein. PMID- 3032111 TI - [Treatment of autochthonous rat brain tumors with chemotherapy and radiotherapy]. AB - The authors tried to establish a model of primary, autochthonous avian sarcoma virus-induced rat glioma for experimental chemotherapy and radiotherapy. It was found that the intracerebral inoculation of 2 X 10(6) FFU/5 microliter of an infectious cells-free homogeneous sub-group D Schmidt-Ruppin avian sarcoma virus into 3-day-old inbred Fischer rats induced brain tumors in all rats. The mean survival time of the inoculated rats was 58.7 +/- 12 days. With regard to the classification of the induced brain tumors in Fischer rats, astrocytoma accounted for 70%. This ASV-induced tumor in rats fulfills the following criteria for a desirable animal model. Spontaneously arising. Glial origin. Intraparenchymal growth. Uniformly fatal within a reasonable time period. In the present study, the therapeutic effects of anticancer drugs, such as ACNU and vincristine were evaluated and additionally, the effect of ACNU used in conjunction with radiation was also evaluated in this model. The mean survival time of rats was prolonged significantly with ACNU (20 mg/kg) or radiation therapy (1,000 rads), respectively, and in cases where ACNU was used together with radiation, the mean survival time was prolonged further still, but not very significantly, in comparison with radiation therapy alone. In conclusion, the ASV-induced rat glioma model was considered to be closely akin to a spontaneous brain tumor in terms of morphology, blood supply and kinetics of the primary tumor. Moreover, the therapeutic sensitivity of this model to anticancer drugs was fairly similar to that of human anaplastic astrocytoma. Considering these observations, this model seems to be an excellent experimental brain tumor model which is useful for evaluating the effect of new therapies against malignant brain tumors. PMID- 3032113 TI - [The efficacy of temporary retention chemotherapy of the treatment of polyps in the remnant rectum of familial polyposis after total colectomy and ileorectostomy -a case report]. AB - A case of familial polyposis is reported. The patient was a 24-year-old woman on whom total colectomy followed by ileorectostomy had been performed. However, there were multiple polyps remaining in the remnant rectum. For the treatment of these residual polyps, a new chemotherapeutic method known as Temporary Retention Chemotherapy (TRC), originally devised by us for the treatment of advanced gastric cancer, was applied. A balloon catheter was inserted and fixed into the rectum via the anus for the treatment of the rectal tumors. Through the catheter, high-dose 5-fluorouracil (5-FU) solution was injected into the rectum and retained there for one hour before being discarded. During the therapy, the patient was asked to change her position of recumbency at certain intervals, so that the 5-FU solution could act evenly on the lesions in question. This therapy was repeated twice a week, and followed by routine endoscopic studies. A total dose of 5,700 mg of 5-FU was necessary for obtaining a complete response of the remnant lesions. 6 months later, 4 polyps were found, and TRC was again applied, resulting in disappearance of the polyps again after the therapy. So far, no side effect have been noted as a result of this therapy. She is currently still alive without any residual polyps, carcinoma or metastasis 6 years after the operation. PMID- 3032114 TI - [FT, 5-FU and uracil concentrations of the blood, bile and tissue of hepatoma with liver cirrhosis after oral administration of UFT]. AB - UFT was orally administered to eight patients with hepatocellular carcinoma (HCC) combined with liver cirrhosis and to five patients with normal liver. The concentrations of FT-207 (FT), 5-FU, and uracil in blood, tissue and bile were then respectively determined. The FT level in cancer tissue and non-cancer tissue was almost identical in patients with HCC. On the other hand, the 5-FU level in normal liver tissue was significantly higher (P less than 0.05) than that in cancer tissue, and the uracil level in normal liver tissue was lower than that in cancer tissue (P less than 0.05). Transportation of FT from blood to liver was significantly correlated with clearance of indocyanine green from the blood (ICGK). These results suggested that transportation of FT from blood to liver and activation of FT were impaired in HCC with liver cirrhosis. However, the 5-FU level in the cancer tissue of HCC tended to be higher than that in non-cancer tissue. The 5-FU level in the tissue had a significant correlation with the FT level in the tissue. In addition, it was presumed that cancer tissue was able to produce 5-FU from FT more quickly than non-cancer tissue. PMID- 3032115 TI - [Activation of macrophage function by OK-432 and its subcellular fractions]. PMID- 3032116 TI - Herpes oesophagitis in a healthy 8 year-old. AB - An 8 year-old, immunocompetent child developed a severe acute herpetic oesophagitis in the absence of oropharyngeal lesions. Intravenous treatment with the antiviral drug, acyclovir, relieved symptoms within 24 hours. PMID- 3032117 TI - Human papillomavirus 25-related DNA in solitary keratoacanthoma. AB - Solitary keratoacanthomas of 32 patients were screened for the presence of human papillomavirus (HPV) 25 DNA, which was originally isolated and molecularly cloned from warts of an epidermodysplasia verruciformis (Ev) patient. Biotinylated virus DNA was hybridized in situ to thin sections obtained from paraffin-embedded material. HPV DNA was detected in 12 of 32 tumors under stringent conditions, and in 2 additional tumors under relaxed conditions. PMID- 3032118 TI - Indigenously acquired human visceral leishmaniasis in Al Agamy (Alexandria Governorate) Egypt. PMID- 3032119 TI - [Prevalence of anti-HIV antibodies in patients without AIDS or AIDS-related syndrome in Kinshasa, Zaire]. PMID- 3032121 TI - [Use of enalapril, an angiotensin-converting enzyme inhibitor, in pulmonary artery hypertension]. AB - Enalapril (E) was used to treat 16 patients with pulmonary arterial hypertension, 6 primary and 10 secondary, 5 of the latter with congenital heart disease and 5 with chronic obstructive pulmonary disease. The average dose of E was 20 mg/day. All patients underwent pre and post-treatment cardiac catheterization with determination of pressures at: right atrial (RA), main pulmonary artery (MPA), pulmonary capillary wedge pressure ( VCP) and systemic arterial (SA). Resistances forces were also measured as; total pulmonary (TPR), pulmonary arteriolar (PAR) and total systemic (TSR) as well as cardiac output (CO), and echo and electrocardiograms, chest x ray, stress test and respiratory function test. The functional class (NYHA) improved in all (p less than 0.001). The initial mean pressures were: RA 12.24 +/- 4.35; MPA 73.81 +/- 25.16; VCP 12 +/- 2.73 and SA 89 +/- 14; TPR 1477 +/- 761; PAR 1243 +/- 730 and TSR 1684 +/- 505.5; CO 4.5 +/- 1.29. The final values were: RA 9.66 +/- 2.46 (p less than 0.001); MPA 63.26 +/- 24.45 (p less than 0.001); VCP 11.33 +/- 2.38 (p = NS); SA 81 +/- 10 (p less than 0.001); TPR 1009.5 +/- 536.7 (p less than 0.001); PAR 829 +/- 511.5 (p less than 0.001); TSR 1309.6 +/- 296.3 (p less than 0.001); CO 5.2 +/- 1.44 (p less than 0.001). The average of minutes on treadmill was initially 8.2 +/- 2.45 and final 12.46 +/- 3.0 (p less than 0.001). It is concluded that enalapril is a useful drug in the treatment of pulmonary arterial hypertension of any etiology. PMID- 3032122 TI - [Use of technetium 99m radio-angiocardiography in the identification of myxoma of the right atrium. Presentation of 2 cases]. AB - A right atrial myxoma was identified in two patients with radio-angiocardiography 99m Tc. Tumor is identified as a persistent photon-deficient region in both first pass and equilibrium images. In addition there is an abnormal accumulation and sluggish disappearance of the radionuclide from the right atrium. Even though sensitivity of the procedure should be confirmed, it is a helpful non-invasive technique of great value in the diagnosis oF right atrial tumors. PMID- 3032120 TI - Magnetic resonance imaging versus computed tomography in the evaluation of soft tissue tumors of the extremities. AB - Twenty patients with extremity soft tissue tumors were prospectively evaluated with magnetic resonance imaging (MRI) and computed tomography (CT) scans with subsequent anatomic correlation of surgical findings. MRI and CT had a similar percentage of accuracy in assessing tumor relationship with major neurovascular (80% and 70%, respectively) and skeletal (80% and 75%, respectively) structures. MRI was significantly better than CT in displaying contrast between tumor and muscle when using the T2 weighted spin echo (SE) (p2 less than 0.002) and inversion recovery (IR) (p2 less than 0.005) pulse sequences. MRI and CT were comparable in demonstrating contrast between tumor and fat. The contrast between tumor and vessel was better displayed by MRI compared with CT when using the T1 weighted SE (p2 less than 0.001) and T2 weighted SE (p2 less than 0.001) pulse sequences. T1 and T2 values were measured on fresh tumor and normal tissue samples and were used to predict relative contrast on different MRI pulse sequences using isosignal contour plots. MRI appears to offer several advantages over CT in the evaluation of extremity soft tissue tumors. PMID- 3032123 TI - [Gamma-angiography during exercise in chronic aortic insufficiency. Development of ventricular function and regurgitation]. AB - Twenty-three patients with chronic aortic incompetence (17 men and 6 women) aged 27 to 71 years (average 51 years) underwent sequential gamma-angiography at rest and during the different levels of exercise and recovery phase to investigate the evolution of ventricular function and regurgitant fraction and so, guide therapy. The radionuclide indices of left ventricular function (end diastolic and end systolic indexed volumes, global ejection fraction, regional wall motion) and the regurgitant fractions were calculated and compared with clinical, echocardiographic, angiographic and haemodynamic data. The changes observed on effort during gamma-angiography allowed identification of 3 groups of patients: Group I: compensated aortic incompetence with a normal left ventricular ejection fraction (0.69 +/- 0.1), a moderate regurgitant fraction (40 per cent +/- 20 per cent) and, during exercise, a stable left ventricular end diastolic volume index (less than 5 per cent variation), an end systolic volume index which decreased (average-13 per cent), an ejection fraction which increases (by more than 0.05 in 62.5 per cent of cases) and with good global and regional wall motion. Group II: intermediate cases with a left ventricular ejection fraction of 0.62 +/- 0.09 and a regurgitant fraction of 60 +/- 16 per cent. Individual variations were observed with this group which either resembled those of Group I or those of Group III.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3032124 TI - Fibroadenomas with stromal cellularity. A clinicopathologic study of 21 patients. AB - Data on clinical and pathologic features of tumors from 21 patients who had breast tumors compatible with juvenile (cellular) fibroadenomas (JCF) were reviewed. All patients were black females ranging in age at time of presentation from 10 to 39 years, with a median age of 15 years. Follow-up (median period, six years) was obtained for 20 patients. Patients were categorized into two groups; 13 had solitary lesions and eight had multiple and successive lesions. No histologic differences were observed between JCF occurring as solitary or multiple tumors or between lesions occurring in patients younger and older than age 20 years. The JCF was shown to be a fairly distinct clinicopathologic entity that characteristically occurred in adolescents, but which was occasionally seen in young adults. It had a benign course, characterized by synchronous and metachronous multicentricity, rather than local recurrence. PMID- 3032125 TI - Immunohistochemical staining for apolipoprotein B in human sarcomas. AB - A study of immunocytochemical staining for apolipoprotein B (apo B) in 52 human sarcomas and eight benign soft-tissue tumors is described. A peroxidase antiperoxidase technique was employed using a rabbit anti-apo B antiserum. All 27 liposarcomas examined were positive for apo B, and 23 of these tumors showed at least 2-plus staining on a scale of 0 to 3 plus. Of the ten myxoid liposarcomas within this group, seven showed 2-plus or 3-plus staining, while the other three stained 1 plus. By contrast, 11 of 13 malignant fibrous histiocytomas were negative for apo B; the remainder showed 1-plus staining. Myxoid areas present in three of these cases were negative for apo B. Five of seven rhabdomyosarcomas stained at least 2 plus for apo B. Both benign and malignant peripheral nerve sheath tumors showed no consistent pattern of staining. We conclude that apo B immunocytochemical staining tends to be high in tumors whose normal tissue counterparts exhibit relatively large numbers of low-density lipoprotein receptors. Furthermore, immunoperoxidase staining for apo B should be a useful adjunct in the evaluation of soft-tissue sarcomas, particularly in cases with myxoid differentiation. PMID- 3032126 TI - Immunoreactive beta-endorphin in ovarian sex cord-stromal tumors. AB - The possible production of the opioid polypeptide beta-endorphin (beta-EP) was investigated in paraffin-embedded tissue from 17 ovarian tumors with the use of a specific anti-beta-EP antibody and the avidin-biotin-peroxidase staining technique. Only sex cord-stromal tumors (ten cases) showed positive staining. Strong beta-EP immunoreactivity was present in Leydig's cells of Sertoli-Leydig cell tumors; weaker sporadic staining was present in cells of granulosa cell tumors, and faint staining was present in occasional, luteinized theca cells of fibrothecomata. These findings suggest that cells with the sex cord-stromal phenotype that are capable of steroid production can also produce beta-EP. The latter may be a component of the "functional" status associated with some ovarian sex cord-stromal tumors and may serve as a helpful marker in distinguishing this type of tumors from germ cell or epithelial neoplasms. PMID- 3032127 TI - Fixation of experimental osteotomies of the distal femur of rabbits with biodegradable material. AB - Osteotomies of the distal femur in 19 rabbits were operatively fixed with totally biodegradable implants. Radiographic, histological, microradiographic, and oxytetracycline-labeling studies showing healing of the osteotomy within 6 weeks. The fixation proved stable sufficiently during healing of the osteotomized bone. The osteotomies united without delay and malalignment did not occur, although no external support was used and the rabbits were allowed to walk freely after the operation. Successful use of the biodegradable fixation developed in the present study was reported recently for the treatment of fractures of the ankle. PMID- 3032128 TI - Constant current sine wave transcutaneous nerve stimulation for the evaluation of peripheral neuropathy. AB - We have evaluated various forms of peripheral neuropathy with a new device which emits a constant sinusoid stimulus at varying frequencies to quantitate current perception thresholds (CPTs). In normal individuals, CPT measures increase with increasing stimulation frequency, are highest on the toe and lowest on the face. There is a significant effect of age and sex on threshold perception. In patients with neuropathy, there is a marked increase in thresholds on hands and feet, as well as a lesser but still significant increase of facial CPTs. Thresholds furthermore correlated with clinical severity in a group of patients with diabetic neuropathy. Facial thresholds were markedly elevated in the patients with moderate to severe neuropathy, suggesting that the device is sensitive to the systemic nature of peripheral neuropathy. The authors believe the device will be a useful tool in screening for sensory neurologic abnormalities. PMID- 3032129 TI - Surgical disease in East Africa. Presidential address. PMID- 3032130 TI - Effects on membrane function by cholesterol oxidation derivatives in cultured aortic smooth muscle cells. AB - Autoxidation derivatives of cholesterol known to affect cholesterol content of the cells were shown to alter some membrane associated functions in cultured aortic smooth muscle cells. For study of membrane-bound enzymes, Na+,K+-ATPase and 5'-nucleotidase were measured cytochemically by electron microscopy. Cells incubated with 10 ug/ml of cholestane-3 beta,5 alpha,6 beta-triol and 25 hydroxycholesterol for 24 to 48 hours showed marked inhibition of both enzyme activities. For study of carrier-mediated hexose transport, radiolabeled 2-deoxy D-glucose was utilized. The uptake of this labeled compound was measured in the cells preincubated with oxidation derivatives of cholesterol for various time periods. Cholestane-3 beta,5 alpha,6 beta-triol had a rapid inhibitory effect on hexose transport, which was reversible after removal of the sterol from the medium. Hexose transport was not significantly altered by 25-hydroxycholesterol after up to 8 hours incubation. Two underlying mechanisms are possible. The prompt onset of the effect of cholestane-3 beta,5 alpha,6 beta-triol may be attributable to an incorporation of the sterol into the cell membranes. On the other hand, 25-hydroxycholesterol, a potent inhibitor of cholesterol biosynthesis, may have a delayed effect on membrane function by depleting the cholesterol available for membrane synthesis. PMID- 3032131 TI - [Combined gastrinoma of the duodenum and glucagonoma of the pancreas]. AB - A female patient, aged 53, with a combined duodenal gastrinoma and pancreatic glucagonoma is reported. Such a combination is an integral part of multiple endocrine neoplasias Type I. In both tumors psammous bodies were found. Gastrinoma was an ulcerogenic tumor resulting in the development of Zollinger Ellison syndrome. Glucagonoma for a long time was asymptomatic. PMID- 3032132 TI - Leukotrienes in aqueous humor of patients with uveitis. PMID- 3032133 TI - Pleomorphic adenoma ('benign mixed tumor') of the lacrimal gland. PMID- 3032134 TI - Localization of Na+, K+-ATPase activity in the tegmentum vasculosum of the chick cochlea. AB - We studied the localization of Na+, K+-ATPase activity in the chick cochlea, utilizing a cytochemical method of K+-p-nitrophenylphosphatase (K+-NPPase), and found that the enzyme activity was limited to the dark cells of the tegmentum vasculosum (TV). Our present results indicate that the dark cells of the TV play an essential role in the maintenance of the ionic composition of the cochlear endolymph. PMID- 3032135 TI - Tissue conditioners as obturators in maxillofacial surgery. PMID- 3032136 TI - Ross River virus infection and epidemic polyarthritis. PMID- 3032137 TI - Clinical experiences with human parathyroid hormone 1-34. AB - Human parathyroid hormone (PTH) 1-34 was given to nine normal subjects and to 10 patients with hypoparathyroidism. There were no side effects associated with the protocol employed. In normal subjects, five statistically significant changes occurred during the period of observation: plasma cyclic adenosine monophosphate (cAMP) rose by a factor of 3 (at 30 min), nephrogenous cyclic AMP rose approximately 40-fold (at 60 min), urinary phosphate rose by a factor of 2 (at 120 min), urine calcium levels fell by 50% between 60 and 120 min, and plasma prolactin rose by a factor of 1.4 (at 60 min). The cAMP responses were significantly blunted in five patients with chronic hypocalcemia, chronic hyperphosphatemia, and detectable serum immunoreactive PTH levels. On the basis of this test these patients were designated as suffering from pseudohypoparathyroidism. The acute phosphaturic and hypocalciuric responses were apparently intact in these five individuals. Human PTH 1-34 is likely to replace bovine material in the delineation of syndromes associated with PTH resistance. PMID- 3032138 TI - Amnesia produced by anisomycin in an appetitive task is not due to conditioned aversion. AB - Two experiments investigated the effects of lithium chloride (LiCl) and anisomycin (ANI) in a water reward Y-maze task. In Experiment 1, male CD-1 mice given weak or strong training were injected post-training with either saline or LiCl (150 mg/kg), which has been reported to produce conditioned aversion in mice. One day after training, both LiCl groups avoided the rewarded arm of the maze and drank less water than saline-injected controls. Two days after training, the strongly trained LiCl mice showed avoidance, while both LiCl groups drank less water. In Experiment 2, weakly trained mice given pre- and post-training ANI (30 mg/kg) were amnesic on the second test day compared to mice that received post-trial saline. However, water consumption was increased on the test day for both groups. LiCl produced a different pattern of results than ANI in this task. On the basis of these results, it is suggested that amnesia produced by ANI is due to impaired memory formation and not to conditioned aversion. PMID- 3032140 TI - The importance of the refractile body in expression of the killer trait in paramecium. PMID- 3032139 TI - Extrachromosomal elements of extrachromosomal elements of Paramecium and their extrachromosomal elements. AB - Expression of killer traits in Paramecium is due to a complex interaction between the lower eukaryote host and two or three elements that can be viewed either as extrachromosomal elements or as endosymbionts. In all cases, the determinants of the killer trait are carried by obligate bacterial endosymbionts belonging to the genus Caedibacter. However, the actual genetic determinants for expression of these traits are not an integral part of the symbiont genome. They are located on extrachromosomal genetic elements (plasmids or bacteriophages) which essentially are molecular endosymbionts of Caedibacter. In the case of the plasmids, they are associated with yet another set of extrachromosomal genetic elements, which are transposons. These transposons have been observed to move into new sites in the plasmids and even to disrupt expression of R body production and the killer trait. Thus, the transposons can be considered either as extrachromosomal elements of extrachromosomal elements (plasmids) of extrachromosomal elements (C. taeniospiralis) of paramecia, or as molecular parasites of molecular endosymbionts (plasmids) of bacterial endosymbionts of paramecia. PMID- 3032141 TI - Mobile elements in the yeast mitochondrial genome. PMID- 3032142 TI - Mitochondrial gene expression in yeast: further studies of a self-splicing group II intron. PMID- 3032143 TI - Structural studies on centromeres in the yeast Saccharomyces cerevisiae. AB - In the yeast Saccharomyces cerevisiae, circular or linear plasmids containing a functional centromere (CEN) and a chromosomal replicator (ARS) are mitotically stable and segregate as ordinary yeast chromosomes in the first and second meiotic divisions. A centromere in S. cerevisiae consists of a region of DNA, approximately 150 bp in length, containing three important sequence elements, which are folded with proteins into a specific conformation in the chromatin (the yeast kinetochore). Each of the functional CEN sequences contains a high (91% to 95%) AT region (element II), 78 to 86 bp in length, flanked on one side by the common sequence PuTCACPuTG (element I), and on the other by the sequence TGTTT.TG.TTTCCGAAA....AAA (element III). Deletions in the element II region partially inactivate mitotic function and cause precocious separation of the sister chromatids in meiosis I. Element III appears to be a protein binding site, as evidenced by the following observations. Various point mutations in element III inactivate centromere function, especially in the central CCG (17). One or more protein binding sites in the element III region can be demonstrated by an exonuclease III blocking assay. Wild-type CEN sequences compete strongly in this binding assay, whereas certain functionally inactive mutant CEN sequences do not. In addition, various DNA segments containing either CEN3 or the element III region strongly repress expression of the yeast GAL1 gene when inserted immediately upstream from the transcriptional start site. Helical DNA segments containing CEN3 or CEN14 are shown to be bent or distorted in shape in the high AT element II region. PMID- 3032145 TI - Regulation of the yeast CYC1 gene. PMID- 3032144 TI - Mitochondrial introns as mobile genetic elements: the role of intron-encoded proteins. AB - Introns of organelle genes share distinctive RNA secondary structures that allow their classification into two known families. These structures are believed to play an essential role in splicing, and members of both structural classes have recently been shown to perform self-splicing reactions in vitro. In lower eukaryotes, many structured introns also contain long internal open reading frames (ORFs), which are able to code for hydrophilic proteins. Several properties of self-splicing structured introns suggest that they resemble mobile genetic elements, even though no actual transposition event involving these introns has yet been found. We report here on the characterization of two intron encoded proteins that strongly support this attractive idea. First, we show that the class I intron of the 21S ribosomal RNA (rRNA) gene of Saccharomyces cerevisiae omega+ strains (rl intron) encodes a specific transposase. This protein has been partially purified from Escherichia coli cells that overexpress it from an artificial universal code equivalent to the rl intronic ORF. The omega transposase shows a double-strand endonuclease activity in vitro. This activity creates a 4-bp staggered cut with 3' OH overhangs within a specific sequence of the 21S rRNA gene of omega- strains. It is precisely within this sequence that the rl intron inserts by a duplicative transposition. Second, we report on the synthesis, in E. coli, of a putative reverse transcriptase encoded by the class II intron of the cytochrome b gene of Schizosaccharomyces pombe. This synthesis was obtained from E. coli expression vectors, using the class II intronic ORF linked to an artificial initiator sequence. As further support of the idea that structured introns are mobile, we show, from a systematic screening of introns in various yeast species, that the rl intron has transposed into the ATPase subunit 9 gene of Kluyveromyces fragilis. Structural features observed at the new intron homing site may be relevant to the transposition event. PMID- 3032146 TI - Translation, post-translational processing, and mitochondrial translocation of yeast iso-1-cytochrome c. PMID- 3032147 TI - Control of yeast gene expression by transposable elements. PMID- 3032148 TI - Mitochondrial plasmids of Neurospora and other filamentous fungi. PMID- 3032149 TI - [Adenoviruses in chickens: an attempt at summarizing]. PMID- 3032150 TI - [The pathogenesis of "egg drop syndrome 76" after experimental infection of one day-old chicks and laying hens]. PMID- 3032151 TI - Tubulin-nucleotide interactions. Effects of removal of exchangeable guanine nucleotide on protein conformation and microtubule assembly. AB - The removal of tightly bound GDP from the exchangeable nucleotide-binding site of tubulin has been performed with alkaline phosphatase under conditions which essentially retain the assembly properties of the protein. When microtubule protein is treated with alkaline phosphatase, nucleotide is selectively removed from tubulin dimer rather than from MAP (microtubule-associated protein) containing oligomeric species. Tubulin devoid of E-site (the exchangeable nucleotide-binding site of the tubulin dimer) nucleotide shows enhanced proteolytic susceptibility of the beta-subunit to thermolysin and decreased protein stability, consistent with nucleotide removal causing changes in protein tertiary structure. Pyrophosphate ion (3 mM) is able to promote formation of normal microtubules in the complete absence of GTP by incubation at 37 degrees C either with nucleotide-depleted microtubule protein or with nucleotide-depleted tubulin dimer to which MAPs have been added. The resulting microtubules contain up to 80% of tubulin lacking E-site nucleotide. In addition to its effects on nucleation, pyrophosphate competes weakly with GDP bound at the E-site. It is deduced that binding of pyrophosphate at a vacant E-site can promote microtubule assembly. The minimum structural requirement for ligands to induce tubulin assembly apparently involves charge neutralization at the E-site by bidentate ligation, which stabilizes protein domains in a favourable orientation for promoting the supramolecular protein-protein interactions involved in microtubule formation. PMID- 3032153 TI - Reaction mechanism of the gastric H+ +K+-dependent ATPase. Effects of inhibitor and pH. AB - The effect of nolinium bromide [2-(3,4-dichlorophenylamino)quinolizium bromide], which acts as a K+ antagonist in the gastric H+ +K+-dependent ATPase reaction, was investigated at the level of 32P-labelled intermediates of the gastric ATPase reaction. A concentration-dependent effect of nolinium bromide was observed on the concentrations of phosphorylated intermediates. At low (up to 50 microM) concentrations the drug did not interfere with the concentrations of intermediates but exhibited a competition with K+ at the level of both 32P labelled intermediates and hydrolysis of ATP at pH 7.0. Similar competition was noted in the H+ +K+-dependent ATPase reaction. Low nolinium bromide concentrations also drastically slowed the enzyme turnover. The concentrations of the intermediates were lowered appreciably between 50 microM- and 100 microM nolinium bromide without affecting the ATP hydrolysis, and the effects were independent of pH. Similar to the effects at pH 7.0, the drug also exhibited competition with K+ in lowering the E approximately P concentration at pH 5.0. A dramatic effect of pH on the K+-sensitivity as well as on turnover of the 32P labelled intermediates was observed. Although the concentrations of intermediates remained nearly unaltered at various pH values, the K+-stimulated hydrolysis of ATP showed an optimum at pH 7.0 with sharp declines at pH 5 and 8. The data suggest a critical involvement of H+ in the conversion of the K+-insensitive E1 approximately P into the K+-sensitive E2 approximately P form of the enzyme. Nolinium bromide appears to function as a K+ analogue and seems to block the entry of K+ at the K X E2 step, thereby interfering with the enzyme turnover. PMID- 3032152 TI - Mechanism of inhibition of mammalian DNA topoisomerase I by heparin. AB - We have previously shown that heparin is a potent inhibitor of a mammalian DNA topoisomerase I. We have now investigated the mechanism of its inhibition. This was carried out first by scrutinizing the structural features of heparin molecules responsible for the inhibition. Commercial heparin preparation was fractionated by antithrombin III-Sepharose into non-adsorbed, low-affinity and high-affinity fractions, of which only the high-affinity fraction of heparin is known to contain a specific oligosaccharide sequence responsible for the binding to antithrombin III. These fractions all exhibited essentially similar inhibitory activities. Furthermore, when chemically sulphated to an extent comparable with or higher than heparin, otherwise inactive glycosaminoglycans such as heparan sulphate, chondroitin 4-sulphate, dermatan sulphate and neutral polysaccharides such as dextran and amylose were converted into potent inhibitors. Sulphated dermatan sulphate, one of the model compounds, was further shown to bind competitively to the same sites on the enzyme as heparin. These observations strongly suggested that topoisomerase inhibition by heparin is attributable primarily, if not entirely, to the highly sulphated polyanionic nature of the molecules. In a second series of experiments we examined whether heparin inhibits only one or both of the topoisomerase reactions, i.e. nicking and re-joining. It was demonstrated that both reactions were inhibited by heparin, but the nicking reaction was more severely affected than was the re-joining reaction. PMID- 3032154 TI - [15N]aspartate metabolism in cultured astrocytes. Studies with gas chromatography mass spectrometry. AB - The metabolism of 2.5 mM-[15N]aspartate in cultured astrocytes was studied with gas chromatography-mass spectrometry. Three primary metabolic pathways of aspartate nitrogen disposition were identified: transamination with 2 oxoglutarate to form [15N]glutamate, the nitrogen of which subsequently was transferred to glutamine, alanine, serine and ornithine; condensation with IMP in the first step of the purine nucleotide cycle, the aspartate nitrogen appearing as [6-amino-15N]adenine nucleotides; condensation with citrulline to form argininosuccinate, which is cleaved to yield [15N]arginine. Of these three pathways, the formation of arginine was quantitatively the most important, and net nitrogen flux to arginine was greater than flux to other amino acids, including glutamine. Notwithstanding the large amount of [15N]arginine produced, essentially no [15N]urea was measured. Addition of NaH13CO3 to the astrocyte culture medium was associated with the formation of [13C]citrulline, thus confirming that these cells are capable of citrulline synthesis de novo. When astrocytes were incubated with a lower (0.05 mM) concentration of [15N]aspartate, most 15N was recovered in alanine, glutamine and arginine. Formation of [6-amino 15N]adenine nucleotides was diminished markedly compared with results obtained in the presence of 2.5 mM-[15N]aspartate. PMID- 3032155 TI - Hexokinase isoenzymes of RIN-m5F insulinoma cells. Expression of glucokinase gene in insulin-producing cells. AB - We have analysed the pattern of expression of the hexokinase isoenzyme group in RIN-m5F insulinoma cells. Three hexokinase forms were resolved by DEAE-cellulose chromatography. The most abundant isoenzyme co-eluted with hexokinase type II from rat adipose tissue and displayed a Km for glucose of 0.15 mM, similar to the adipose-tissue enzyme. Hexokinase type II was in large part associated with a particulate subcellular fraction in RIN-m5F cells. The two other hexokinases separated by ion-exchange chromatography were an enzyme similar to hexokinase type I from brain and glucokinase (or hexokinase type IV). The latter isoenzyme was identified as the liver-type glucokinase by the following properties: co elution with hepatic glucokinase from DEAE-cellulose and DEAE-Sephadex; sigmoid saturation kinetics with glucose with half-maximal velocity at 5.6 mM and Hill coefficient (h) of 1.54; suppression of enzyme activity by antibodies raised against rat liver glucokinase; apparent Mr of 56,500 and pI of 5.6, as shown by immunoblotting after one- and two-dimensional gel electrophoresis; peptide map identical with that of hepatic glucokinase after proteolysis with chymotrypsin and papain. These data indicate that the gene coding for hepatic glucokinase is expressed in RIN-m5F cells, a finding consistent with indirect evidence for the presence of glucokinase in the beta-cell of the islet of Langerhans. On the other hand, the overall pattern of hexokinases is distinctly different in RIN-m5F cells and islets of Langerhans, since hexokinase type II appears to be lacking in islets. Alteration in hexokinase expression after tumoral transformation has been reported in other systems. PMID- 3032156 TI - Cytochrome c protein-synthesis rates and mRNA contents during atrophy and recovery in skeletal muscle. AB - It is known that immobilization of the rat hindlimb by plaster casting leads to muscle atrophy and loss of muscle protein. In the present study, immobilization of the rat hindlimb for 6 h resulted in a significant 27% decrease in the absolute rate of cytochrome c synthesis in the red quadriceps muscle, without any change in the relative amount of cytochrome c mRNA. Cytochrome c mRNA in normal red quadriceps muscle was observed to be of four different lengths (1400, 1050, 650 and 580 bases). After 7 days of immobilization, the absolute rate of cytochrome c synthesis remained depressed and cytochrome c mRNA decreased by 40%; each of the cytochrome c mRNAs decreased, with a preferential disappearance of the 1050- and 1400-base lengths. Immobilization was ended on day 7, and the atrophied muscle was allowed to recover. At day 4 of recovery, the absolute rate of cytochrome c synthesis was 92% higher and the amount of cytochrome c mRNA had returned to control values. The abundances of the 1050- and 1400-base cytochrome c mRNAs had increased more than the shorter cytochrome c mRNAs, so that they were higher than control values. It appears that acute decreases in contractile activity of the red quadriceps muscle alter cytochrome c synthesis rates via translational or post-translational mechanisms, whereas chronic periods of modified contractile activity alter its synthesis rate via pre-translational mechanisms. PMID- 3032158 TI - Regulation of 3-hydroxy-3-methylglutaryl-coenzyme A reductase activity in mouse uterine epithelial cells. AB - The regulation of 3-hydroxy-3-methylglutaryl-CoA reductase was studied in mouse uterine epithelium. The enzyme was rapidly inactivated during incubation with ATP/Mg2+ in vitro, and could be re-activated by incubation with partially purified rat liver phosphoprotein phosphatase. Enzyme activity was rapidly inhibited by mevalonate injection in vivo to approx. 30% of control. The percentage of total enzyme active in vivo was measured by inclusion of NaF in the isolation buffers. The percentage of enzyme active in vivo 18 h after stimulation by oestrogens remained at approx. 25% after inhibition of activity by mevalonate injection, cholesterol feeding or progesterone pretreatment. However, 9 h after oestrogen stimulation, cholesterol feeding inhibited enzyme activity to 57% of control, 94% of which was in the active form. We conclude that, although all components for a reversible phosphorylative regulation of 3-hydroxy-3 methylglutaryl-CoA reductase activity are present in uterine epithelial cells, a role in the rapid changes in epithelial enzyme activity has not been demonstrated. PMID- 3032157 TI - Superoxide-dependent and ascorbate-dependent formation of hydroxyl radicals from hydrogen peroxide in the presence of iron. Are lactoferrin and transferrin promoters of hydroxyl-radical generation? AB - Apo-lactoferrin and apo-transferrin protect against iron-ion-dependent hydroxyl radical (.OH) generation from H2O2 in the presence of superoxide radicals or ascorbic acid at pH 7.4, whether the necessary iron is added as ionic iron or as ferritin. Iron-loaded transferrin and lactoferrin [2 mol of Fe(III)/mol] show no protective ability, but do not themselves accelerate .OH production unless chelating agents are present in the reaction mixture, especially if the proteins are incorrectly loaded with iron. At acidic pH values, the protective ability of the apoproteins is diminished, and the fully iron-loaded proteins can release some iron in a form able to accelerate .OH generation. The physiological significance of these observations is discussed. PMID- 3032159 TI - The circular-dichroic properties of the 'Rieske' iron-sulphur protein in the mitochondrial ubiquinol: cytochrome c reductase. AB - We have studied the c.d. spectra of the 'Rieske' iron-sulphur protein isolated from the ubiquinol: cytochrome c reductase (bc1 complex) of bovine heart mitochondria. Both the oxidized and the reduced form of the 'Rieske' protein display a series of well-resolved c.d. features resembling those reported for the 'Rieske'-type iron-sulphur protein purified from the bacterium Thermus thermophilus [Fee, Findling, Yoshida, Hille, Tarr, Hearshen, Dunham, Day, Kent & Munck (1984) J. Biol, Chem. 259, 124-133]. In particular, the difference spectra, reduced minus oxidized, of both proteins have a distinctive negative band at 497 nm. The c.d. features characteristic of the isolated 'Rieske' protein were found in the dichroic spectra of the whole bc1 complex in the region between 450 and 520 nm. The reduction of the enzyme by ascorbate or ubiquinol is accompanied by the formation of a negative band at about 500 nm that corresponds, in all its c.d. properties, to the specific dichroic absorption of the reduced 'Rieske' iron sulphur protein. PMID- 3032160 TI - Maternal administration of dexamethasone stimulates choline-phosphate cytidylyltransferase in fetal type II cells. AB - Administration of dexamethasone to pregnant rats at 19 days gestation increased phosphatidylcholine synthesis (45%) from radioactive choline in type II cells. This enhanced synthesis of phosphatidylcholine was accompanied by an increased conversion of choline phosphate into CDP-choline. Similar results were obtained by incubating organotypic cultures of 19-day-fetal rat lung with cortisol. The increased conversion of choline phosphate into CDP-choline correlated with an enhanced choline-phosphate cytidylyltransferase activity (31% after dexamethasone treatment; 47% after cortisol exposure) in the cell homogenates. A similar increase (26% after dexamethasone treatment; 39% after cortisol exposure) was found in the microsomal-associated enzyme. No differences in cytosolic enzyme activity were observed. The specific activity of the microsomal enzyme was 3-4 times that of the cytosolic enzyme. Most of the enzyme activity was located in the microsomal fraction (58-65%). The treatments had no effect on the total amount of enzyme recovered from the cell homogenates. These results, taken collectively, are interpreted to indicate that the active form of cytidylyltransferase in type II cells is the membrane-bound enzyme and that cytidylyltransferase activation in type II cells from fetal rat lung after maternal glucocorticoid administration occurs by binding of inactive cytosolic enzyme to endoplasmic reticulum. PMID- 3032163 TI - Isolation and purification of chloroplastic spinach (Spinacia oleracea) sedoheptulose-1,7-bisphosphatase. AB - Higher-plant sedoheptulose-1,7-bisphosphatase was isolated and purified over 200 fold from spinach (Spinacia oleracea) chloroplast stromal extracts to apparent electrophoretic homogeneity by DEAE-Fractogel, molecular sieving on Sephadex G 200 and Blue B dye-matrix affinity chromatography. It is a protein of Mr 66,000, made up of two apparently identical subunits (Mr 35,000). The enzyme is activated by reduced thioredoxin fb in the presence of dithiothreitol. Its specificity towards sedoheptulose 1,7-bisphosphate versus fructose 1,6-bisphosphate is high, but not absolute. PMID- 3032164 TI - Maturation of rabbit reticulocytes: strong decline of the turnover of polyphosphoinositides. AB - The [32P]phosphate labelling of ATP and polyphosphoinositides (PI) of intact rabbit reticulocytes and mature erythrocytes was compared. Despite the fact that reticulocytes have a 30-fold higher turnover of ATP than mature erythrocytes, the 32P-labelling of the cellular ATP pool was identical. The 32P-Pi entry into the cells is the rate-limiting step for 32P-phosphate isotopic equilibration of the ATP-pool and is independent of maturation. The rates of 32P-phosphate incorporation into PI-4,5-P2 and into PI-4-P, respectively, were 17-fold and 8 fold higher in reticulocyte-rich cell suspensions than in erythrocytes. The specific radioactivity of PI-4,5-P2 labelling reaches 55% and that of PI-4-P 40% of the ATP specific activity. However, a rapid isotopic equilibration of the polyphosphoinositides, was found. These findings indicate the existence of a heterogeneity of polyphosphoinositide pools. Reticulocyte membranes lose about 30% of their polyphosphoinositides during maturation proportional to the total loss of phospholipids. PMID- 3032161 TI - The leukotriene B4 paradox: neutrophils can, but will not, respond to ligand receptor interactions by forming leukotriene B4 or its omega-metabolites. AB - Leukotriene B4 (5S,12R-dihydroxy-6,14-cis,8,10-trans-eicosatetraenoic acid, LTB4) is released from neutrophils exposed to calcium ionophores. To determine whether LTB4 might be produced by ligand-receptor interactions at the plasmalemma, we treated human neutrophils with serum-treated zymosan (STZ), heat-aggregated IgG and fMet-Leu-Phe (fMLP), agonists at the C3b, Fc and fMLP receptors respectively. STZ (10 mg/ml) provoked the formation of barely detectable amounts of LTB4 (0.74 ng/10(7) cells); no omega-oxidized metabolites of LTB4 were found. Adding 10 microM-arachidonate did not significantly increase production of LTB4 or its metabolites. Addition of 50 microM-arachidonate (an amount which activates protein kinase C) before STZ caused a 40-fold increase in the quantity of LTB4 and its omega-oxidation products. Neither phorbol myristate acetate (PMA, 200 ng/ml) nor linoleic acid (50 microM), also activators of protein kinase C, augmented generation of LTB4 by cells stimulated with STZ. Neither fMLP (10(-6) M) nor aggregated IgG (0.3 mg/ml) induced LTB4 formation (less than 0.01 ng/10(7) cells). Moreover, cells exposed to STZ, fMLP, or IgG did not form all-trans-LTB4 or 5-hydroxyeicosatetraenoic acid; their failure to make LTB4 was therefore due to inactivity of neutrophil 5-lipoxygenase. However, adding 50 microM arachidonate to neutrophil suspensions before fMLP or IgG triggered LTB4 production, the majority of which was metabolized to its omega-oxidized products (fMLP, 20.2 ng/10(7) cells; IgG, 17.1 ng/10(7) cells). The data show that neutrophils exposed to agonists at defined cell-surface receptors produce significant quantities of LTB4 only when treated with non-physiological concentrations of arachidonate. PMID- 3032162 TI - Purification and characterization of a major glycoprotein in rat hepatoma plasma membranes. One of the membrane proteins released by phosphatidylinositol-specific phospholipase C. AB - A major glycoprotein of rat hepatoma plasma membranes was selectively released as a soluble form by incubating the membrane with phosphatidylinositol-specific phospholipase C. The soluble form corresponding to the glycoprotein was also prepared by butan-1-ol extraction of microsomal membranes at pH 5.5, whereas extraction at pH 8.5 yielded an electrophoretically different form with a hydrophobic nature. The soluble glycoprotein extracted at pH 5.5 was purified by sequential chromatography on concanavalin A-Sepharose, Sephacryl S-300 and anti (alkaline phosphatase) IgG-Sepharose, the last step being used to remove a contaminating alkaline phosphatase. The glycoprotein thus purified was a single protein with Mr 130,000 in SDS/polyacrylamide-gel electrophoresis, although it behaved as a dimer in gel filtration on Sephacryl S-300. The glycoprotein was analysed for amino acid and carbohydrate composition. The composition of the carbohydrate moiety, which amounted to 64% by weight, suggested that the glycoprotein contained much larger numbers of N-linked oligosaccharide chains than those with O-linkage. It was confirmed that the purified glycoprotein was immunologically identical not only with that released by the phospholipase C but also with the hydrophobic form extracted with butan-1-ol at pH 8.5. The results indicate that the glycoprotein of rat hepatoma plasma membranes, which has an unusually high content of carbohydrate, is another membrane protein released by phosphatidylinositol-specific phospholipase C, as documented for alkaline phosphatase, acetylcholinesterase and Thy-1 antigen. PMID- 3032166 TI - Slime moulds and the origin of foldback DNA. PMID- 3032165 TI - [Establishment and characterization of the pig aortic endothelial cell line BSEz 3 in a serum-reduced medium]. AB - The cell line BSEz-3 was established in a serum-reduced medium MEMPAS. The cells were isolated and cultivated under the same conditions as the calf aortic endothelial cell line BKEz-7. With regard to cell isolation and cell density in the stationary phase, BSEz-3-cells show characteristic differences from BKEz-7 cells. For the isolation of BSEz-3-cells an enzymatic-mechanical method was more successful than a mechanical technique only. The low saturation density in the stationary phase with an average number of 50,000 cells/cm2 of glass surface area is considered as a strong contact inhibition of cell proliferation. As a cell type specific marker the BSEz-3-cells contain factor-VIII-antigen and possess angiotensin-converting enzyme activity. For a practical use of the BSEz-3-cells in drug research we give recommandations regarding cell inoculum density for subcultivation and cryo-conservation in ampoules. PMID- 3032168 TI - Growth factors and their receptors: specific roles in development. PMID- 3032167 TI - Viral sequences required for neurovirulence of poliovirus. PMID- 3032169 TI - ATP stimulates inositol phosphates accumulation and calcium mobilization in a primary culture of rat aortic myocytes. AB - The effects of extracellular ATP on phosphoinositide metabolism and intracellular Ca2+ concentration were studied in a primary culture of rat aortic myocytes. ATP increases the level of inositol phosphates, the putative second messenger for Ca2+ mobilization. No saturation of inositol phosphates accumulation is obtained (up to 10(-2) M ATP). Under the same conditions, ATP rapidly mobilizes intracellular Ca2+ in fura-2 loaded myocytes. The mobilization of intracellular Ca2+ is dose-dependent (maximal at 10(-4) M ATP), and is not affected by addition of EGTA. It is concluded that the receptors mediating the cytosolic increase of Ca2+ are of the P2-purinoceptor subtype. The physiological functions of these receptors are not presently known. PMID- 3032170 TI - G-protein dissociation, GTP-GDP exchange and GTPase activity in control and PMA treated neutrophils stimulated by fMet-Leu-Phe. AB - The addition of the chemotactic factor fMet-Leu-Phe to cell homogenates causes a decrease in the pertussis toxin catalyzed ADP-ribosylation of a 41 kDa protein. The fMet-Leu-Phe induced decrease is not abolished in homogenates prepared from phorbol 12-myristate 13-acetate treated neutrophils. This decreased ribosylation probably reflects a dissociation of the GTP-binding protein oligomer that is not followed by association, possibly because of the release of the alpha-subunit into the suspending medium. Furthermore, fMet-Leu-Phe stimulates the binding of radiolabelled guanylylimidodiphosphate to membrane preparations. Again, the stimulated binding of guanylylimidodiphosphate is not affected by treating the intact neutrophils with phorbol 12-myristate 13-acetate. In addition leukotriene B4, platelet activating factor and fMet-Leu-Phe activate a high-affinity GTPase in membrane preparations. The basal level of this GTPase activity is dramatically inhibited in membrane preparations isolated from cells treated with phorbol 12 myristate 13-acetate. On the other hand, the fMet-Leu-Phe stimulated component is only marginally reduced. The present findings suggest that PMA does not prevent receptor G-protein interaction. PMID- 3032171 TI - Partially purified phosphatidylinositol kinase does not catalyze the formation of phosphatidylinositol-4,5-bisphosphate. AB - The enzyme phosphatidylinositol kinase was partially purified from murine livers. The purification scheme involved solubilization of proteins with Triton X-100 and deoxycholate, followed by gel filtration chromatography in ACA 44, affinity chromatography with Blue Sepharose and hydroxylapatite. The purification achieved from membranes was 490 fold. We found that partially purified phosphatidylinositol kinase was unable to catalyze the formation of phosphatidylinositol-4,5-bisphosphate. PMID- 3032172 TI - Retinoic acid alters the metabolic 3H-labelling of glycosphingolipids. AB - Retinoic acid increases the incorporation of radioactivity from a mixture of [3H] galactose and [3H]-glucosamine into glycosphingolipids of serum-starved quiescent human foreskin fibroblasts with a preferential labelling of ceramide mono- and dihexoside as compared to ceramide tri- and tetrahexoside. Under the conditions used, no similar change in the specific labelling of glycoprotein is observed. Alteration in [3H]-precursor uptake into glycolipids comparable to that seen under the influence of retinoic acid does not occur in the presence of phorbolester, colchicine, butyrate or after infection with cytomegalovirus. PMID- 3032173 TI - Physiological correlation between nucleoside-diphosphate kinase and the enzyme associated guanine nucleotide binding proteins. AB - The physiological correlation between NDP-kinase and the enzyme-associated guanine nucleotide binding proteins (G1 and G2) has been studied in vitro. It was found that incubation of the phosphoenzyme (enzyme-bound high-energy phosphate intermediate) of NDP-kinases with one of the nucleoside 5'-diphosphates (NDPs) in the presence of divalent cations (Mg2+ and Ca2+) results in the formation of nucleoside 5'-triphosphates (NTPs) within 40 sec even at low temperatures (below 4 degrees C) without strict base-specificity; and high-energy phosphates on the phosphoenzyme can transfer preferentially to GDP on the guanine nucleotide binding proteins (G1, G2 and r-p21 protein) in the presence of 0.25 mM Ca2+ or 1 mM Mg2+ even if any other NDPs are present in the reaction mixtures. These observations suggest that NDP-kinase may be responsible for the phosphate transfer between GDP on the guanine nucleotide binding proteins and its phosphoenzyme. PMID- 3032175 TI - Effects of diadenosine 5',5'''-P1, P4-tetraphosphate and of adenosine 5' triphosphate on the higher order structure of calf thymus DNA. AB - The effective length and the hard core radius were calculated by scaled particle theory for high molecular weight calf thymus DNA in the presence of varying concentrations of diadenosine 5',5'''-P1, P4-tetraphosphate and of adenosine 5' triphosphate in aqueous millimolar NaCl. DNA became slightly more flexible in the presence of diadenosine 5',5'''-P1, P4-tetraphosphate at concentrations of 10(-9) 10(-7) M. DNA was denatured in the presence of 5 X 10(-5) M adenosine triphosphate. PMID- 3032174 TI - Monoclonal anti-parathyroid antibodies interfering with a Ca2+-sensor of human parathyroid cells. AB - Previous findings indicate that binding of Ca2+ to an external receptor is part of the mechanism by which extracellular Ca2+ regulates cytoplasmic Ca2+ and hormone release of parathyroid cells. We now present evidence that two newly generated monoclonal anti-parathyroid antibodies react with structures involved in this sensing and/or gating of Ca2+. Microfluorimetric studies of fura 2-loaded human parathyroid cells thus revealed that the antibodies competed with Ca2+ and antagonized the rise in cytoplasmic Ca2+ normally obtained when parathyroid cells are exposed to increasing concentrations of extracellular Ca2+. PMID- 3032177 TI - Enzymatic hydrolysis of leukotriene A4 into 5,6-dihydroxy-7,9,11,14 eicosatetraenoic acid and LTB4 by mammalian kidney. AB - Homogenates from rat and pig kidney converted leukotriene A4 to 5,6-dihydroxy 7,9,11,14-eicosatetraenoic acid as well as leukotriene B4. Both hydrolyses were enzymatic as judged by the effects of heat treatment and proteolytic digestion. Upon subcellular fractionation, conversion of leukotriene A4 to 5,6-dihydroxy 7,9,11,14-eicosatetraenoic acid occurred both in the 105,000xg supernatant and the 20,000xg pellet from rat kidney, whereas conversion to leukotriene B4 was confined to the 105,000xg supernatant. We also found production of 5,6-dihydroxy 7,9,11,14-eicosatetraenoic acid and leukotriene B4 in isolated rat renal epithelial cells, either from exogenous leukotriene A4 or from this substrate supplied by human leukocytes. PMID- 3032176 TI - The diacylglycerol kinase inhibitor, R59022, enhances the superoxide generation from human neutrophils induced by stimulation of fMet-Leu-Phe, IgG and C3b receptors. AB - A specific diacylglycerol kinase inhibitor, at a concentration of 10(-5) M, consistently enhanced superoxide generation from human neutrophils stimulated with fMet-Leu-Phe, IgG, heat-aggregated IgG and opsonized zymosan. The concentration-response curve for fMet-Leu-Phe was displaced to the left and the maximum superoxide release was also consistently increased by R59022 whereas the diacylglycerol lipase inhibitor, RHC80267, 10(-5) M, had no significant effect. These results suggest that the diacylglycerol formed after fMet-Leu-Phe stimulation in human neutrophils is metabolized largely by the kinase and not the lipase, which implies that diacylglycerol is not the major source of arachidonate during signal-transduction. PMID- 3032178 TI - Activation of interleukin-2 receptor gene by forskolin and cyclic AMP analogues. AB - Forskolin (FK), a reversible activator of adenylate cyclase, markedly enhanced the expression of interleukin-2 receptor (IL-2-R) on a human natural killer (NK) like cell line, YT. The FK-induced increase in IL-2-R on YT cells closely correlated with an increase in intracellular cyclic AMP (cAMP) level, and was mimicked by dibutyryl cyclic AMP (dbcAMP). FK induced both high and low affinity IL-2-R on the cells. Using a cDNA for the IL-2-R as a probe, the FK-induced IL-2 R expression was shown to be associated with an increase in IL-2-R mRNA. FK also enhanced the IL-2-R expression on a human T lymphotrophic virus I (HTLV-I) positive T-cell line (YTA-1H) and augmented the phorbol ester-induced expression of IL-2-R on HTLV-I negative T-cell lines (HSB-2 and HPB-ALL). These results suggest the possibility that the stimulation of adenylate cyclase may serve as a pathway leading to activation of the IL-2-R gene in certain types of lymphocytes. PMID- 3032180 TI - Transcriptional induction of proto-oncogene fos by HSV-2. AB - HSV-2 induces the transcription of c-fos gene in both human (diploid) cell strains and mouse (heteroploid) cell lines. HSV-1 is incapable of such induction. The induction of transcription is rapid and transient in both cell types. The product of transcriptional induction of c-fos in mouse cells is mainly a 4.5 kb fos mRNA along with lesser amount of a 2.2 kb fos mRNA, while in human cells the 4.5 kb mRNA is the only detectable product. Both the 4.5 kb and 2.2 kb fos mRNAs are mature gene products as judged by their presence exclusively in the cytoplasm of infected mouse L929 cells, while a 3.5 kb putative precursor of the 2.2 kb fos mRNA appears exclusively in the nucleus. In infected human WI38 cells, the 4.5 kb mRNA also appears exclusively in the cytoplasm. Virus specific protein synthesis is an absolute requirement for the transcriptional activation. The 4.5 kb fos mRNA is under different control mechanisms since it is not induced by serum stimulation of serum starved cells as is the 2.2 kb fos transcript. Moreover, in serum starved cells simultaneously stimulated by serum and infected, only the 2.2 kb fos transcript can be superinduced by the protein synthesis inhibitor cycloheximide, while the appearance of the 4.5 kb fos mRNA is completely blocked by this treatment. PMID- 3032179 TI - Fluorescein mercuric acetate specifically displaces zinc from cytochrome oxidase. AB - Treatment of beef heart cytochrome oxidase with fluorescein mercuric acetate (FMA) was found to specifically displace zinc from the enzyme and inhibit the steady-state activity in a parallel fashion. The native cytochrome oxidase preparation contained 2.3 Cu: 2.0 Fe: 1.1 Zn: 0.9 Mg. Addition of 2 equivalents of FMA inhibited the activity by 50% and displaced 60% of the zinc from the enzyme, but did not affect the copper, iron or magnesium content. The pre-steady state reduction of cytochrome oxidase by ferrocytochrome c was not affected by the FMA treatment, in contrast to the inhibition of steady state activity. These results suggest a possible structural or functional role for zinc in cytochrome oxidase. PMID- 3032181 TI - A 2-deoxy analogue of KDO as the first inhibitor of the enzyme CMP-KDO synthetase. AB - The two KDO analogues 2,6-anhydro-3-deoxy-D-glycero-D-galacto-octonate and 2,6 anhydro-3-deoxy-D-glycero-D-talo-octonate were synthesized and tested as inhibitors of the enzyme CTP:CMP-deoxyoctulosonate cytidylyltransferase (CMP-KDO synthetase) from Gram-negative bacteria. Only compound 4, the 2-deoxy analogue of beta-KDO-pyranose, was found to be an inhibitor with a Ki of 3.9 microM. PMID- 3032182 TI - Adenosine 3',5'-cyclic monophosphate stimulates secretion of alpha-melanocyte stimulating hormone from permeabilized cells of the intermediate lobe of the rat pituitary gland. AB - The effect of Mg-ATP and cyclic AMP on the secretion of alpha-melanocyto stimulating hormone (alpha-MSH) from electrically permeabilized cells of rat intermediate lobe (IL) were investigated. Addition of exogenous Ca2+ stimulated alpha-MSH secretion in a concentration- (EC50 = 4.8 microM) and temperature dependent manner. This Ca2+-evoked secretion was further enhanced by Mg-ATP and cyclic AMP. Mg-ATP was required for the fully secretory response in the electrically permeabilized IL cells and the maximal secretion was reached at 1 mM. Cyclic AMP in the presence of GTP gamma S also potentiated Ca2+-evoked alpha MSH secretion to the same magnitude as Mg-ATP. In the absence of Ca2+ both the cyclic AMP and Mg-ATP did not stimulate alpha-MSH secretion from IL cells. The data suggest that Mg-ATP and cyclic AMP may modulate directly the secretory components rather than change intracellular concentration of free Ca2+. PMID- 3032183 TI - Isolation of a cDNA clone for rat liver 6-phosphofructo 2-kinase/fructose 2,6 bisphosphatase. AB - A cDNA clone for rat liver 6-phosphofructo 2-kinase/fructose 2,6-bisphosphatase was isolated from a lambda gt11 rat liver expression library by antibody screening. The clone was approximately 1100 bases in length and the derived amino acid sequence contained 303 residues at the carboxyl end of the subunit. This derived amino acid sequence corresponded exactly with the actual amino acid sequence of the enzyme determined by direct sequencing of the protein. PMID- 3032184 TI - beta-Galactosidase from rat epididymal fluid is bound by a recognition site attached to membranes of the epididymis different from the phosphomannosyl receptor. AB - In order to know if the beta-galactosidase of the rat epididymal fluid, as other secreted acid hydrolases, carries a marker in its molecule, we studied the binding of this enzyme to cellular membranes of the epididymal tissue. The binding, like that mediated by the phosphomannosyl receptor, was saturable, did not require calcium, had a Kd in the nM range and was inhibited by phosphatase or metaperiodate treatment of the enzyme. However fructose 6-phosphate derivates were more effective competitive inhibitors than mannose 6-phosphate. The binding capacity of the membranes were extractable with Triton X-100 and incorporable into liposomes. Trypsin inhibited the binding capacity of Triton extracts but it did not affect the affinity of intact cellular membranes for beta-galactosidase. The results suggest that a phosphorylated carbohydrate of the enzyme is bound by a recognizing site of the cellular membranes different from the phosphomannosyl receptor. PMID- 3032185 TI - Conservation of E6 and E7 regions of human papillomavirus types 16 and 18 present in cervical cancers. AB - The E6 and E7 regions of human papillomavirus (HPV) type 16 were present in the DNA samples from cervical cancer cell lines, SKG-IIIa and SKG-IIIb, and those from cervical cancer tissues of three different patients. T601 cells, an NIH3T3 transformant obtained by transfection of DNA from a surgical specimen of a cervical cancer, also contained the E6 and E7 regions. The E6 region of HPV type 16 was expressed as mRNA in SKG-IIIa, SKG-IIIb and T601 cells. The E6 and E7 regions of HPV type 18 were present in the DNA samples from cervical cancer cell lines, SKG-I and SKG-II, and those from cervical cancer tissues of two different patients. SKG-I and SKG-II cells expressed the E6 region of HPV type 18 as mRNAs. These results strongly suggest that the E6 and E7 regions or the sequence surrounding these regions are important for maintaining malignant phenotype of cervical cancer cells. PMID- 3032186 TI - Vitamins E and K induce aryl hydrocarbon hydroxylase activity in human cell cultures. AB - Two fat soluble vitamins, Vitamins E and K, when added into culture medium, were found to increase aryl hydrocarbon hydroxylase activity in human cultured cells. The extent of induction in a hepatoma-derived cell line (Hep G2) by these vitamins is of similar magnitude to those cells receiving benz[a]anthracene; whereas in a mammary tumor-derived cell line (MCF-7), benz[a]anthracene is the best inducer for the hydroxylase activity. The increase of the hydroxylase activity is associated with increased levels of a specific mRNA coding for polynuclear aromatic hydrocarbons-induced form of cytochrome P-450 with Vitamins E and K treatment. The size of the induced mRNA is 3.3 kilobase which is the same as that of benz[a]anthracene treatment. PMID- 3032187 TI - LH-releasing activity and receptor binding of pHGnRH 14-26 analogues. AB - The human gonadotropin-releasing hormone (GnRH) precursor consists of the GnRH sequence followed by a cleavage and amidation site and a 56-amino acid carboxyl terminal extension (pHGnRH - precursor human GnRH) which has been shown to stimulate gonadotropin release. This activity has been localized to a decapeptide sequence (corresponding to pHGnRH 17-26) in its amino-terminal region using human pituitary cell cultures. To further characterize the structural features required for gonadotropin release, two analogues, [D-Ala17]pHGnRH 14-26 and [D Trp22]pHGnRH 14-26, with D-amino acid substitutions inside and peripheral to this decapeptide sequence were chemically synthesized. pHGnRH 14-26 and the D-Ala17 analogue were inactive and GnRH, pHGnRH 14-36 and the D-Trp22 analogue stimulated luteinizing hormone release from cultured rat pituitary cells in a calcium dependent, dose-responsive manner. Experiments and receptor binding studies with the active pHGnRH peptides in conjunction with GnRH or a GnRH antagonist suggest that the active pHGnRH peptides act through the GnRH receptor. PMID- 3032188 TI - Isolation of a biotin receptor from hepatic plasma membranes. AB - Biotin, one of the growth promoting members of the B complex vitamin family, was found in the present investigation to have a specific receptor on isolated plasma membranes. Biotin bound to its receptor in a linear manner at 20 degrees C with some binding as early as 30 minutes and full equilibrium binding being present at 20 hours. The best binding was found at 0.6 mg/ml of protein, but significant binding was still present at 0.6 microgram/ml. The saturation of ligand binding sites was at 10(-7)M. Half maximal saturation of binding was between 10(-9) and 10(-10)M. These results demonstrate that a receptor for biotin does exist on purified plasma membranes. PMID- 3032189 TI - The effect of polymyxin B nonapeptide (PMBN) on transformation. AB - The effect of the outer membrane permeabilizing polycation, polymyxin B nonapeptide (PMBN) on the transformation of E. coli HB101 with pBR322 plasmid DNA was investigated. Pretreatment of cells with PMBN (followed by suspending the cells in PMBN-free medium) did not stimulate the development of competence induced by the calcium heat pulse. In the absence of calcium-ions, a high PMBN concentration (1 mM) was able to induce a low transformation frequency provided that PMBN was not removed before the addition of DNA. PMID- 3032190 TI - Spin trapping artifacts in DMSO. AB - Spin-trapping experiments in alkaline aqueous dimethyl sulfoxide (DMSO) solution using sodium 3,5-dibromo-4-nitrosobenzenesulfonate (DBNBS) yielded a strong signal of the sulfur trioxide anion radical adduct. This radical adduct is identical to that obtained by the oxidation of sulfite with horseradish peroxidase/hydrogen peroxide and subsequent spin trapping with DBNBS. This radical adduct is very stable, and satellite peaks of the natural abundance 13C and 33S could be obtained. Apparently, under alkaline conditions DMSO decomposes in air to form the sulfur trioxide anion radical. A comparison with a recent publication shows that this DMSO-derived radical adduct has been misassigned as a uniquely stable spin adduct of superoxide (Ozawa and Hanaki (1986) Biochem. Biophys. Res. Commun. 136, 657-664). PMID- 3032191 TI - Glycerol metabolism in higher plants: glycerol kinase. AB - Glycerol kinase activity was identified in extracts of higher plant seeds and seedlings, and was partially purified and characterized from cucumber radicle tissue. The enzyme was localized in the post-mitochondrial supernatant of the cell, and catalyzed the formation of glycerol-3-phosphate. The pH optiumum was 9.0. ATP, CTP, GTP or UTP could be used as the phosphoryl group donor. The Km for glycerol was 55 microM and Km values for the nucleoside triphosphates were 145 620 microM. The Vmax for the reaction was 40-78 pmol product per min. Kinetic data indicate that the enzyme has a sequential mechanism. PMID- 3032192 TI - Acetaldehyde-mediated hepatic lipid peroxidation: role of superoxide and ferritin. AB - Evidence in alcoholics as well as in experimental models support the role of hepatic lipid peroxidation in the pathogenesis of alcohol-induced liver injury, but the mechanism of this injury is not fully delineated. Previous studies of the metabolism of ethanol by alcohol dehydrogenase revealed iron mobilization from ferritin that was markedly stimulated by superoxide radical generation by xanthine oxidase. Peroxidation of hepatic lipid membranes (assessed as malondialdehyde production) was studied during in vitro alcohol metabolism by alcohol dehydrogenase. Peroxidation was initiated by acetaldehyde-xanthine oxidase, stimulated by ferritin, and inhibited by superoxide dismutase or chelation or iron with desferrioxamine. In conclusion, lipid peroxidation may be initiated during the metabolism of ethanol by alcohol dehydrogenase by an iron dependent acetaldehyde-xanthine oxidase mechanism. PMID- 3032193 TI - Arachidonic acid inhibits phosphate transport by cultured renal cells. AB - Because arachidonic acid and its metabolites are reported to be intracellular messengers of various exogenous stimuli, we studied whether arachidonic acid influences phosphate transport by cultured mouse renal epithelial cells. Arachidonic acid, at 10(-7)-10(-4)M, inhibited phosphate transport without influencing cyclic adenosine 3':5'-monophosphate production. Nordihydroguaiaretic acid and indomethacin, inhibitors of arachidonic acid metabolism, did not cancel the arachidonic acid-induced inhibition of phosphate transport. Furthermore, unsaturated fatty acids other than arachidonic acid also inhibited phosphate transport and their inhibitory effect increased as the number of double bond increased. These data demonstrate that arachidonic acid inhibits the phosphate transport by the cultured renal epithelial cells, probably not via conversion to its metabolites. PMID- 3032194 TI - High-performance liquid chromatography using novel square tile-shaped hydroxyapatite crystals as adsorbent. AB - High-performance liquid chromatography using, as adsorbent, novel square tile shaped hydroxyapatite crystals (with thicknesses of about 2 microns and diameters of 3-7 microns) has been developed. The chromatographic efficiencies of the novel hydroxyapatite packed columns are almost equal to those of the previously developed spherical hydroxyapatite packed columns; high chromatographic resolutions can be obtained by using extremely reduced column lengths of 0.5-3 cm. Since both the square and the spherical hydroxyapatite have roughly the same particle size of some micrometers, the chromatographic efficiency can be deduced to be determined mainly by the particle size rather than the particle shape. PMID- 3032195 TI - Adriamycin-stimulated hydroxyl radical formation in human breast tumor cells. PMID- 3032196 TI - Inhibition by p-nitrophenylphosphate of acetylcholine release induced by Na+ deprivation. AB - The effect of p-nitrophenylphosphate (p-NPP) on the release of acetylcholine evoked by drugs and ionic environments known to inhibit Na+, K+-ATPase was studied in isolated cortical slices of rat brain and longitudinal muscle strip of guinea-pig ileum. p-NPP inhibited the release of acetylcholine induced by sodium deprivation provided that the circumstances were in favour of the function of the K+-activated part of ATPase. However, it failed to antagonize the increase in the acetylcholine release elicited by omission of K+ or by administration of ouabain. Therefore it is concluded that the K+-stimulated phosphatase moiety of the Na+, K+-ATPase might be involved in the release of acetylcholine. PMID- 3032197 TI - Induction of cytosolic and microsomal epoxide hydrolases and proliferation of peroxisomes and mitochondria in mouse liver after dietary exposure to p chlorophenoxyacetic acid, 2,4-dichlorophenoxyacetic acid and 2,4,5 trichlorophenoxyacetic acid. AB - The effects of dietary exposure to 0.125% (w/w) p-chlorophenoxyacetic acid, 2,4 dichlorophenoxyacetic acid or 2,4,5-trichlorophenoxyacetic acid on the content of peroxisomes and levels of certain xenobiotic-metabolizing enzymes in mouse liver have been investigated. In agreement with the literature on rat liver 2,4 dichlorophenoxyacetic acid and 2,4,5-trichlorophenoxyacetic acid were found to cause extensive proliferation of peroxisomes (as judged by the total levels of "mitochondrial" protein, carnitine acetyltransferase, cyanide-insensitive palmitoyl-CoA oxidation and catalase) in mouse liver. On the other hand, exposure to p-chlorophenoxyacetic acid did not significantly affect any of these parameters. As with certain other peroxisome proliferators, 2,4 dichlorophenoxyacetic acid and 2,4,5-trichlorophenoxyacetic acid increased total cytochrome oxidase activity as well. In addition, dietary exposure to 2,4 dichlorophenoxyacetic acid and to 2,4,5-trichlorophenoxyacetic acid resulted in increases in the activities of cytosolic and microsomal epoxide hydrolases in mouse liver and generally less pronounced increases in the total cytosolic glutathione transferase activity and microsomal content of cytochrome P-450. In the case of cytochrome P-450, this process can be said to be a true induction (i.e. the amount of enzyme protein is increased), because the assay procedure for cytochrome P-450 measures holoenzyme amount. Immunoquantitation demonstrated that this was also the case for the changes in cytosolic epoxide hydrolase. The dramatic differences in proliferation of peroxisomes and induction of xenobiotic metabolizing enzymes seen here with compounds differing relatively little in structure may indicate that a receptor mechanism of some kind is involved. PMID- 3032198 TI - Lymphotoxic activity of methyl prednisolone in vitro--I. Comparative toxicity of methyl prednisolone in human cell lines of B and T origin. AB - The cytotoxic activity of methyl prednisolone was compared in EB-3(B), NALM-6(B), CCRF-CEM(T) and RPMI-8226 (plasma cell) cell lines derived from human lymphoid malignancies. Whereas EB-3 cells were steroid-sensitive, NALM-6 cells were partially sensitive and CCRF-CEM and RPMI-8226 were steroid resistant at concentrations of methyl prednisolone up to 10(-4) M. A high concentration of methyl prednisolone, 2.5 X 10(-3) M was toxic to all cell lines. Steroid sensitivity did not correlate with the incorporation of [3H] dexamethasone and could not be mimicked by flurbiprofen, a non-steroidal anti-inflammatory agent. Both theophylline and di-butyryl cAMP were toxic towards NALM-6, EB-3 and CCRF CEM cells; however, this toxicity was reversible and did not reflect the cells' sensitivities towards methyl prednisolone. Furthermore, elevated levels of cAMP in theophylline-treated cells, were not demonstrable in cells treated with methyl prednisolone at toxic or non-toxic concentrations of the steroid. Steroid sensitive EB-3 cells exposed to 10(-5) M methyl prednisolone, produced a soluble factor which was toxic CCRF-CEM cells. PMID- 3032200 TI - Pitfalls in demonstrating an endogenous ligand of imipramine recognition sites. AB - The recognition sites for the 5-hydroxytryptamine (5-HT) uptake inhibitors imipramine and paroxetine may represent receptors for a presently unknown endogenous ligand, whose function would be to modulate 5-HT uptake. Attempts to isolate such a factor from rat brain tissue are described, following a published procedure. It is shown that chromatographic fractions found to inhibit the binding of [3H]imipramine and [3H]paroxetine to rat brain membranes consisted of material essentially unretained by the reverse-phase HPLC column, and they were of high osmolarity. Thus, the activity was probably unspecific in nature, and the presence in rat brain of the factor has not been unequivocally demonstrated. PMID- 3032199 TI - Comparison of the stimulation of inositol phospholipid hydrolysis and of cyclic GMP formation by neurotensin, some of its analogs, and neuromedin N in neuroblastoma clone N1E-115. AB - Neurotensin, some of its analogs, and neuromedin N were examined for comparison of their potencies at stimulating inositol phospholipid hydrolysis and cyclic GMP synthesis in intact murine neuroblastoma cells (clone N1E-115). Neurotensin(8-13) and acetylneurotensin(8-13) had the highest potencies for the stimulation of the hydrolysis of inositol phospholipid, which were about three times as potent as neurotensin (EC50 = 0.9 nM). On the other hand, fragments of the amino-terminal portion of neurotensin, such as neurotensin(1-6), neurotensin(1-8) and neurotensin(1-11), showed no ability to stimulate this hydrolysis. Neuromedin N, which is similar in structure to neurotensin(8-13) and which has been demonstrated to stimulate cyclic GMP formation [J.A. Gilbert and E. Richelson, Eur. J. Pharmac. 129, 379 (1986)], had EC50 values of 2.5 and 4.5 nM for release of [3H]inositol phosphates and stimulation of cyclic [3H]GMP respectively. A strong correlation was obtained between the EC50 values for neurotensin and several analogs in the stimulation of the release of inositol phosphates and the EC50 values for these peptides in the stimulation of cyclic GMP formation in neuroblastoma clone N1E-115 cells under similar experimental conditions. Thus, these two different biochemical effects of neurotensin and its analogs appear to be mediated by the same receptor site, which may also have been the site of action of neuromedin N in these cells. PMID- 3032201 TI - Cyclic AMP efflux from rat striatal slices is enhanced by CCK. PMID- 3032202 TI - [3H]-nitrendipine binding in membranes obtained from hypoxic and reoxygenated heart. AB - We compared the binding properties of [3H]-nitrendipine in heart membranes from normal guinea-pig heart and from hypoxic or hypoxic and reoxygenated heart. The [3H]-nitrendipine binds a single class of high capacity (Bmax 667.2 +/- 105.2) with high affinity (KD 0.14 +/- 0.02) binding sites. By contrast, in membranes of hypoxic and reoxygenated heart the Bmax decreases significantly while it remains unaffected during hypoxia. Xanthinoxidase activity is increased in hypoxic reoxygenated hearts. PMID- 3032203 TI - Photosensitization of SV 40 DNA mediated by promazine derivatives and 4' hydroxymethyl-4,5',8-trimethylpsoralen. Inhibition of the in vitro transcription. AB - In vitro transcription by E. coli RNA polymerase was carried out on SV40 DNA photoreacted with various promazine derivatives. Inhibition of the template activity was recorded with increasing irradiation times in the presence of promazine derivatives. Promazine covalent adducts on guanine did not terminate RNA synthesis and seemed to be bypassed by the enzyme. HMT (4'-hydroxymethyl 4,5',8-trimethylpsoralen) photoreaction with DNA was carried out under two conditions: irradiation with lambda greater than 395 nm favouring monoadduction on pyrimidine residues and irradiation at 360 nm inducing a maximum of interstrand diadducts. Both adducts were able to terminate RNA synthesis on the phototreated SV40 DNA and using the O-methyl-nucleotide sequencing procedure, the termination sites were precisely mapped. Monoadducts on the coding strand and cross-links induced termination two bases away from the covalent adduct, but monoadducts on the noncoding strand did not half RNA polymerase. PMID- 3032204 TI - The amplified chemiluminescence test to characterize antirheumatic drugs as oxygen radical scavengers. AB - High levels of reactive oxygen species (ROS) are generated by phagocytes involved in host defence and inflammation. Thus, it appears highly desirable to learn more about the potential of antirheumatic drugs to scavenge ROS or to inhibit their enzymatic generation. Amplified chemiluminescence (CL) allows detection of O-2 using lucigenin (LgCL) or H2O2 using luminol (LuCL). A total of 43 compounds have been tested quantitatively in vitro (10(-6) to 10(-4) mol/l) with respect to three test parameters; varying cell-activity, and incubation-time employing two different phagocyte populations (neutrophils/macrophages). The most active compounds with LgCL were the known radical scavengers nordihydroguaiaretic acid (NDGA), N-propyl gallate, superoxide dismutase and chloroquine, the non-steroidal anti-inflammatory drugs (NSAID) benzydamine, timegadine, carprofen, enolicam, the known lipoxygenase inhibitors (e.g. CBS 1108/1114, BW 755C) and glucosaminoglucan polysulfate. Inactive in this system were corticosteroids (prednisolone, dexamethasone) most of the tested NSAID (N = 16/20), most disease modifying drugs (D-penicillamine, levamisole, gold-TM) and the anti-gout drugs (sulfinpyrazone, allopurinol, colchicine). Therefore amplified CL with lucigenin appears to be a rapid, kinetic, reproducible means of pharmacological profiling in vitro new anti inflammatory drugs for radical scavenger activity. PMID- 3032205 TI - Evaluation of membrane fluidity effects and enzyme activities alterations in adriamycin neurotoxicity. AB - Adriamycin (ADR) increased the lipid fluidity of dog brain synaptosomal plasma membranes (SPM) labeled with 1,6-diphenyl-1,3,5-hexatriene (DPH), as indicated by the steady-state fluorescence anisotropy [(ro/r)-1]-1. Arrhenius-type plots of [(ro/r)-1]-1 indicated that the lipid phase separation of the membrane at 23.3 +/ 1.2 degrees was perturbed by ADR such that the temperature was reduced to 16.2 +/- 1.1 degrees. Arrhenius plots of (Na+ + K+)-stimulated ATPase activity exhibited a break point at 22.8 +/- 1.1 degrees in control SPM which was reduced to 15.8 +/- 1.0 degrees in ADR treated SPM, suggesting differences in the interaction of (Na+ + K+)-stimulated ATPase with lipids between ADR treated and untreated SPM. (Na+ + K+)-stimulated ATPase and Ca2+-stimulated ATPase activities were increased at a concentration range 10(-18)-10(-15) M of ADR; higher concentrations (up to 10(-7) M), however, led to a progressive inhibition of the enzyme activities. The allosteric properties of SPM-bound (Na+ + K+)-stimulated ATPase by fluoride (F-) (as reflected by changes in the Hill coefficient) were modulated by ADR whereas those of SPM-bound acetylcholinesterase remained unaffected. We propose that ADR achieves these effects through asymmetric perturbations of the membrane lipid structure and that changes in membrane fluidity may be an early key event in ADR induced neurotoxicity. PMID- 3032206 TI - The effects of hydrazine on the phosphatidate phosphohydrolase activity in rat liver. AB - Injection of hydrazine (0.7 mmole/kg) in the male fasting rats caused an increase in phosphatidate phosphohydrolase (PAP) activity in the soluble fraction of the liver. The increased PAP activity was parallel with a rise in hepatic triacylglycerol (TG) (3.5-fold) and in the catecholamine concentration (3.4-fold) in adrenal glands. Hydrazine also increased serum glucose. The hydrazine-induced increase in PAP activity and TG accumulation was completely prevented by adrenalectomy. The data suggest that increased PAP activity is at least partly responsible for hydrazine-induced fatty liver and that adrenal hormones may take part in the mechanism by which hydrazine exerts its effects on the liver. PMID- 3032207 TI - Is histamine potentiation of adenosine-stimulated cyclic AMP accumulation in guinea-pig cerebral cortical slices mediated by products of inositol phospholipid breakdown? PMID- 3032208 TI - [Primary structure of the alpha-subunit of Na+,K+-ATPase from the swine kidney. III. Complete nucleotide sequence corresponding to the structural region of the gene]. AB - The nucleotide sequence of the cDNA, containing coding region of the alpha subunit of the pig kidney Na+, K+-ATPase, was determined. The region contains 3063 b.p. coding for 1021 amino acid residues. In the course of processing, five amino acid residues are cleaved to yield the mature Na+, K+-ATPase alpha-subunit containing 1016 amino acid residues. PMID- 3032209 TI - [Primary structure of the beta-subunit of Na+,K+-ATPase from the swine kidney. I. Analysis of products of trypsin hydrolysis of immobilized proteins]. AB - The Na+, K+-ATPase's beta-subunit immobilized on thiol-glass was hydrolyzed with trypsin. Over 25 peptides covering ca. 90% of the protein polypeptide chain were isolated from the digest by HPLC and characterized. Structural analysis allowed us to localize the sites of attachment of all three carbohydrate chains of beta subunit. Sequence data were used to design of oligonucleotide hybridization probes for gene cloning. PMID- 3032210 TI - Low plasma androgens in women with systemic lupus erythematosus. AB - The high ratio of women with systemic lupus erythematosus (SLE) has remained unexplained, despite the recent description of metabolic abnormalities of estrogen and androgen metabolism. Alterations of steroid metabolism in patients with SLE could be important in the pathogenesis of this disease, since it has been reported that gonadal steroids modulate the immune system. Moreover, research with inbred lupus mice has shown that estrogens have adverse effects on the disease in both sexes, whereas androgen therapy or oophorectomy is protective in females. Recently, the finding of elevated testosterone oxidation at C-17 in females with SLE suggested that plasma androgen levels in males and females with SLE should be examined more closely. We studied the varying degrees of clinical activity, with regard to plasma levels of 4 significant androgens: testosterone, androstenedione, dehydroepiandrosterone, and dehydroepiandrosterone sulfate in a series of 5 male and 42 female SLE patients. Decreased levels of all androgens were observed in women with SLE. The lowest levels of plasma androgens were found in female patients who had active disease, as determined by laboratory and clinical assessment. These data support the fact that specific abnormalities of androgen metabolism in the female are associated with SLE, and may contribute in some way to morbidity and mortality. More importantly, these data may have implications for future therapeutic regimens based on male hormone replacement. PMID- 3032211 TI - Modulation of prostaglandin E2 synthesis in rabbit synoviocytes. AB - Prostaglandin E2 (PGE2) production by rabbit synoviocytes was markedly stimulated by hydroxyapatite only after a 60-minute delay. Release of 3H-arachidonic acid (C20:4) and 3H-PGE2 from cells with phospholipids that were prelabeled with 3H C20:4 did not occur in parallel. At 240 minutes, phospholipase activity in sonicated cell suspensions had increased only 38%, while cyclooxygenase activity had doubled (109%). This doubling, as well as the production of synoviocyte PGE2, was prevented by inhibiting the synthesis of protein. Cyclooxygenase is an inducible enzyme, and as such, it is a rate-controlling step in PGE2 production. PMID- 3032212 TI - Usefulness of pimobendan in the treatment of heart failure. AB - The effects of the benzimidazole-pyridazinone pimobendan (UD-CG 115 BS) on systemic haemodynamics, myocardial performance and the distribution of cardiac output were studied in open-chest anaesthetized pigs. After intravenous bolus injections (0.1-0.5 mg X kg-1, n = 7) increases in heart rate (up to 37%), LVdP/dtmax (up to 54%) and decreases in systemic vascular resistance (up to 33%) and left ventricular filling pressure (up to 50%) were observed, while cardiac output was unchanged. Vasodilation occurred in nearly all regional vascular beds, but was most pronounced in the adrenals (200%), followed by stomach (150%), small intestines (130%), heart (125%) and brain (110%). O2-consumption was not affected in spite of the increases in heart rate and myocardial inotropy. To evaluate the direct effects on the myocardial, pimobendan was also infused (1-5 micrograms X kg-1 X min-1, n = 7) directly into the left anterior descending coronary artery. In addition to a marked vasodilation of the coronary bed (140%), also a lowering of the left ventricular filling pressure (up to 20%) and cardiac output (15%) was observed, but no changes in regional myocardial function, LVdP/dtmax and systemic vascular resistance occurred. Immediately after intracoronary bolus injections (1 mg X kg-1, n = 4), vasodilation of the coronary vessels was apparent, but myocardial contractility was not affected. This may explain that cyclic AMP content, determined in biopsies excised 30 s after injection, was unaltered. It may be concluded that pimobendan exerts actions on the cardiovascular system which may be useful in the treatment of heart failure. PMID- 3032213 TI - Peptide vaccine--a new approach to a safer foot-and-mouth disease virus vaccine. AB - A synthetic peptide, of which the region of the major antigenic determinant of foot-and-mouth disease virus serotype O1K located on the coat protein VP1 consists, was coupled to different protein carriers. Comparing the potency of the conjugates to elicit neutralising antibodies it has been shown that KLH was the best carrier protein. Using different amounts of peptide A (aa 144-aa 159) the dependence of neutralising antibody response on the amount of injected peptide has been demonstrated. Peptide A was shown to be located in the C-terminal part of a highly variable region when the protein sequences of different sero- and subtypes were compared. Comparative evaluation of peptide A with two longer peptides G1-21 and G1-31 (aa 140-aa 160 and aa 129-aa 160) suggests that the N terminal part of the highly variable region before amino acid 144 is not necessary to obtain a protective response, but it is necessary to enhance the immune response when the peptides are used uncoupled. PMID- 3032214 TI - Analysis of bacterial exopolysaccharides. AB - Extracellular polysaccharides have been isolated from cultures of freshwater and marine bacteria originally isolated from material adhering to surfaces and underivatized hydrolysates have been analyzed by high-performance liquid chromatography methods. A scheme has been developed whereby the uronic acids can be identified on strong anion-exchange columns, while neutral monosaccharides can be separated and identified using aminobonded columns or cation-exchange adsorbent loaded with a heavy metal ion. The methods permit rapid and accurate comparison of polysaccharides with differing chemotype. The strains studied show a range of different chemotypes, all containing a uronic acid and several neutral monosaccharides. Some of the polysaccharides isolated from marine bacteria possessed a very high acetyl content. PMID- 3032215 TI - Cell fusion by nasopharyngeal carcinoma-derived Epstein-Barr virus. AB - Cell fusion experiments using Epstein-Barr virus (EBV) obtained from nasopharyngeal carcinoma (NPC) hybrid cells (NPC-KT) were carried out to examine whether human adenoid epithelial cells (Ad-AH) could be fused with lymphoblastoid cells (BJAB, Raji, and A2L) superinfected or infected by EBV, and if Ad-AH nuclei of the fused cells could express EBV-associated nuclear antigen. The presence of NPC EBV could induce EBV early antigen in all three lymphoblastoid cell lines. By autoradiography, we found that NPC EBV could induce cell fusion between Ad-AH cells and tritiated thymidine-labeled lymphoblastoid cells. In addition, we found that Ad-AH nuclei of the fused cells expressed EBV-associated nuclear antigen four days after NPC EBV-mediated cell fusion of Ad-AH cells with lymphoblastoid cells. PMID- 3032216 TI - Antigenic analysis of testicular cytochromes c using monoclonal antibodies. AB - Testicular cytochrome c (cyt ct) was isolated from testes of sexually mature, rat, mouse, rabbit, and bull, among which rat testis is highly rich in cyt ct. By fusion of NS-1 myeloma cells and spleen cells of mice immunized with rat cyt ct, 11 stable mouse hybridoma cell lines were established. Using an enzyme-linked immunosorbent assay, it was determined that 4 of the 11 anti-rat cyt ct monoclonal antibodies (McAb) did not bind to somatic cyt c (cyt cs) of vertebrates nor to cyt ct of mouse, rabbit, and bull. Four other McAb showed no binding to cyt cs but showed different patterns of cross-reactivity with these four cyt ct. Therefore, these McAb appear to be very sensitive and useful probes for the discrimination or identification of extremely similar isocytochromes c. Although the primary amino acid sequences between cyt cs of rat and mouse are identical, the antigenic structure of cyt ct of rat and mouse are clearly distinct with regard to cross-reactivity with some anti-rat cyt ct McAb. Furthermore, these McAb also reveal that the primary amino acid sequences of cyt ct, which reflect differences in the surface conformation of the molecule, are probably species specific. PMID- 3032217 TI - The relative degradation of [14C]eicosapentaenoyl and [3H]arachidonoyl species of phosphatidylinositol and phosphatidylcholine in thrombin-stimulated human platelets. AB - The platelet-rich plasma from volunteers who had consumed a supplement containing eicosapentaenoate (EPA) was incubated with [3H]arachidonic acid (AA) and [14C]EPA so as to provide for the labelling of these fatty acids in the individual platelet phospholipids. Washed dual-labelled platelet suspensions were prepared and incubated with and without thrombin in the presence of BW755C and in the presence and absence of trifluoperazine (TFP) or indomethacin. The platelet lipids were extracted and the individual phospholipids, as well as diacylglycerol and phosphatidic acid, were separated by thin-layer chromatography and the radioactivity in each fraction was determined. The [3H]AA/[14C]EPA dpm ratio for the loss of radioactivity from phosphatidylcholine (PC) upon thrombin stimulation, as well as that in the residual PC remaining after stimulation, was similar to that in PC in the resting platelets. This suggests no marked selectivity in the degradation of EPA-versus AA-containing species of PC during platelet activation. The [3H]/[14C] ratios for the increased radioactivity appearing in diacylglycerol (DG) and phosphatidic acid (PA) upon thrombin stimulation were not significantly different from the corresponding ratio in phosphatidylinositol (PI) from resting platelets, suggesting little or no preference for 1-acyl-2-eicosapentaenoyl PI over 1-acyl-2-arachidonoyl PI in the pathway from PI to DG to PA. These results suggest that the relative formation of the 2- and 3-series prostaglandins, including thromboxane (Tx) A2 and A3, in stimulated platelets is not regulated by a preferential loss of one of the corresponding eicosanoid precursors over the other from membrane PC and PI. PMID- 3032218 TI - Structure of the O-chain of the lipopolysaccharide of Haemophilus pleuropneumoniae serotype 1. AB - By phenol-water extraction an aqueous-phase soluble cellular lipopolysaccharide was isolated from Haemophilus pleuro-pneumoniae serotype 1. It was shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, hydrolysis, methylation, and both one- and two-dimensional 1H and 13C nuclear magnetic resonance studies to be an S-type lipopolysaccharide, which could be cleaved to yield a lipid A and an O-chain polysaccharide identified as a high molecular weight branched polymer of a tetrasaccharide repeating unit having the structure: (Formula: see text). PMID- 3032219 TI - Restriction of single-stranded M13 DNA using synthetic oligonucleotides: the structural requirement of restriction enzymes. AB - A targeted ss (single stranded) DNA cleavage technique is reported which involves the use of synthetic oligomers complementary to the ss M13 DNA polylinker. BamHI, SmaI, and KpnI restriction enzymes were tested with a partial duplex DNA formed from ss M13 DNA and a nested series of fragments derived from a synthetic 21-mer which were complementary to the polylinker region. These enzymes require up to two flanking nucleotides in addition to the hexameric recognition site for efficient cleavage. This technique could be useful for effecting unique cleavages of DNA with enzymes which generally give a large number of fragments and for strategies of ss DNA manipulation. PMID- 3032220 TI - Changes of cAMP and cGMP levels of rat antral and fundic gastric mucosa in different ulcer models. AB - In both ulcer models investigated, except the 240-min IND and 8-hr STR cAMP values in the antrum, the cyclic nucleotides showed a significant decrease in the gastric mucosa, which is most probably a sign of cellular exhaustion. PMID- 3032221 TI - Na+/K+-ATPase activity of different types of striated muscles. AB - Na+/K+-ATPase activity was determined in striated muscles with different aerobic capacities. The underlying hypothesis was that different aerobic capacities are reflective of different contractile activity which imposes greater demands on sarcolemmal ion translocation and may thus set Na pumping capacity. The added ion translocation demands required during exercise-training on Na+/K+-ATPase activity in different muscle fiber types may require an adaptation of this enzyme. The highest and lowest Na+/K+-ATPase activity was in the heart and white gastrocnemius muscle (WG), respectively. A high linear correlation existed between Na+/K+-ATPase activity and succinate dehydrogenase activity in the six muscles studied. Exercise-training did not increase Na+/K+-ATPase activity in any of the muscles, but did increase the aerobic capacity, except in the heart and WG. It was concluded that Na+/K+-ATPase activity has a high positive correlation with the aerobic capacity of striated muscles in the rat and that the Na pump capacity does not adapt to exercise-training of 1 hr X day-1 as does aerobic capacity. PMID- 3032222 TI - Dot immunodetection for sphingomyelinase with monoclonal antibody. AB - An assay for the detection of sphingomyelinase with monoclonal antibodies is described. The assay takes advantage of nitrocellulose membranes as antigen adsorbent on which a dilute sample can be concentrated as a spot, using a specially designed 96-well filtration device which is commercially available. The method requires only 6 micrograms of the extracts from leukocytes and liver, and it is 10 times more sensitive than the colorimetric assay. This reduced amount of sample material also has the merit of requiring only a 0.5-ml blood sample from patients. The combination of this dot immunoassay with the monoclonal antibody allows a sensitive and a specific assay, and is also applicable as a screening test on a large number of samples. Furthermore, the possibility of differential diagnosis of Niemann-Pick disease types by detecting isoenzymes by this method was examined after isoelectric focusing of placental sphingomyelinase. PMID- 3032223 TI - Lipolysis and beta-adrenergic receptor binding on adipocytes of spontaneously hypertensive rats. AB - Adipocytes from spontaneously hypertensive rats demonstrated a blunted lipolytic response to isoproterenol and dibutyryl cyclic AMP. (-)-[3H]Dihydroalprenolol binding was examined in adipocytes from normotensive and spontaneously hypertensive rats. Increasing concentrations of isoproterenol decreased total (-) [3H]dihydroalprenolol binding to intact cells from normotensive rats, and the efficacy of competition was decreased in adipocytes from spontaneously hypertensive rats. Scatchard analysis indicated that the number of (-) [3H]dihydroalprenolol binding sites and the affinity of dihydroalprenolol binding were comparable between normotensive and spontaneously hypertensive rats. Isoproterenol- and Gpp(NH)p-stimulated adenylate cyclase activity was consistently depressed in adipocyte membranes from spontaneously hypertensive rats as compared to normotensive rats. No difference in fluoride-stimulated adenylate cyclase activity was observed. The blunted lipolytic and cyclic AMP response to isoproterenol in these cells suggest a postreceptor lesion of the lipolytic pathway (possibly the guanine nucleotide regulatory protein) in adipocytes from spontaneously hypertensive rats. The blunted lipolytic response to dibutyryl cyclic AMP suggests defective regulation of lipolytic enzymes at the protein kinase-hormone-sensitive lipase level. PMID- 3032224 TI - Use of 99Tcm radionuclides to show nephrotoxicity of cyclosporin A in transplanted kidneys. AB - In 60 patients, 99Tcm pertechnetate or 99Tcm methylene diphosphonate was injected through an arm vein within 36 h after renal transplantation and repeated 24 or 48 h later. Thirty-one were treated with azathioprine and prednisone, 27 of the grafts had a well-defined peak on the histograms similar to that from the iliac artery histograms at both examinations. In contrast, of the 29 patients treated with cyclosporin A, only 5 of the grafts had a well-defined peak at both examinations. The difference is highly significant. Treatment with cyclosporin A causes a decrease in renal blood flow in the days immediately after transplantation. PMID- 3032225 TI - Platelet alpha-adrenoceptors--a valid model for brain or vascular adrenoceptors? PMID- 3032227 TI - Peripheral neutrophil inclusions in amiodarone treated patients. AB - In 14 patients receiving chronic amiodarone therapy the appearance of multilamellar inclusion bodies in peripheral blood neutrophils was related to both plasma concentrations of amiodarone and its desethyl metabolite and unwanted effects of the drug. Seven of the patients had well defined inclusion bodies. In this group mean amiodarone and desethylamiodarone concentrations were significantly higher than in the remaining seven patients and all but one had unwanted effects of the drug. Of the seven patients without inclusion bodies only one, with high plasma amiodarone and desethylamiodarone concentrations, had unwanted effects of the drug. It is concluded that the appearance of multilamellar bodies in the blood neutrophils of amiodarone-treated patients may help to distinguish those patients at risk of long-term amiodarone toxicity. PMID- 3032228 TI - Is there any interaction between digoxin and enalapril? PMID- 3032226 TI - Topical steroid treatment reduces arachidonic acid and leukotriene B4 in lesional skin of psoriasis. AB - Topical clobetasol propionate or vehicle ointment was applied daily for 3 days to psoriatic plaques on eight patients. Skin chamber exudates from untreated, steroid and vehicle treated lesions were assayed for arachidonic acid (AA), leukotriene B4 (LTB4), prostaglandin E2 (PGE2) and 12-hydroxy-5,8,10,14 eicosatetraenoic acid (12-HETE) before, and at 24 h and 72 h after treatment. Significant reductions in AA and LTB4 were observed at 72 h in steroid treated lesions. The reduction in 12-HETE levels observed after steroid treatment was not statistically significant. PGE2 levels in lesional psoriatic skin were unaltered. The reduction of AA, and LTB4 was associated with clinical improvement of psoriasis. PMID- 3032229 TI - Hereditary spherocytosis revealed by human parvovirus infection. PMID- 3032230 TI - Some peculiarities of the pulmonary phagocytotic response: dust retention kinetics and silicosis development during long term exposure of rats to high quartz dust levels. AB - Rats were exposed to quartz dust (about 90 mg/m3) for five hours a day, five times a week either throughout the 48 weeks of the experiment or for a total of 40 weeks plus eight weeks of "rest." Cytological study of bronchoalveolar lavage showed that at a certain level of silicotic changes in the lungs, a pronounced breakdown in pulmonary dust clearance by macrophages could be observed. There was, however, a concomitant compensatory increase in the contribution to pulmonary phagocytosis by the neutrophil leukocytes (NL). As a result, pulmonary dust kinetics could be mathematically described using a constant clearance rate both throughout the 48 weeks of exposure and during the eight weeks after the end of the 40 week exposure. It is shown in a separate experiment that enhanced recruitment of NL into the airways may be due not only to an attractant effect of the products of macrophage breakdown (PMB) themselves but also to the release of an NL attractant factor by viable macrophages activated under influence of the PMB. PMID- 3032232 TI - Chrysotile asbestos exposure and mesothelioma. PMID- 3032231 TI - Sister chromatid exchanges in rat pleural mesothelial cells treated with crocidolite, attapulgite, or benzo 3-4 pyrene. PMID- 3032233 TI - The influence of guar-gum bread on the regulation of diabetes mellitus type II in elderly patients. AB - The effect of a high-fibre bread mixed with guar-gum (75 g/kg flour) on serum glucose, connecting peptide (C-peptide), haemoglobin A1 (HbA1), high-density lipoprotein-cholesterol (HDL-cholesterol) and triglycerides was examined in fourteen elderly patients with diabetes mellitus type II. The mean daily consumption of guar gum was 8.1 g. Gastrointestinal disorders were not observed. Consumption of guar-gum bread resulted in a significant decrease in C-peptide values on the 1st day and blood glucose values after 3 weeks (both measured 90 min after breakfast). C-peptide values remained low, while an unaccountable 'rebound' phenomenon was seen in the blood glucose values 90 min after breakfast at the end of the study. No significant change was seen in respect to HDL cholesterol and triglycerides. However, a small significant increase in HbA1 was noted. PMID- 3032234 TI - The effect of prolonged dietary supplementation with guar gum on subsequent iron absorption and retention in rats. AB - The effect of prolonged consumption of guar gum on iron absorption and Fe status was investigated in rats. Experiments with closed loops of duodenum, isolated in situ, were designed to reveal changes in the short-term regulation of duodenal Fe uptake, induced by challenge with low- and high-Fe meals. In separate experiments, the effect of guar gum on the capacity of intact rats to maintain Fe status and to absorb Fe from a test-meal was investigated. Male Wistar rats were given either a control, semi-synthetic diet (C) for 21 d or a similar diet containing 100 g guar gum (G)/kg for 27 d. Both diets contained 36 mg Fe/kg. Two subgroups were then challenged with meals containing low-Fe (8 mg/kg) or high-Fe (566 mg/kg), while a third subgroup received a meal of the control diet (36 mg Fe/kg). At intervals of 12, 36, 60 and 84 h after the dietary challenge, the uptake of [59Fe]ferric citrate was measured using closed duodenal loops in situ. All G-supplemented animals absorbed less Fe than their C-fed counterparts. Within group C, animals given the high-Fe challenge had lower absorptions 12, 36 and 60 h later, compared with those given the maintenance diet, whilst those given the low-Fe meal showed much increased uptake 12 and 36 h later. The latter effect was virtually abolished by guar gum. In the second experiment, male Wistar rats were fed on the C or G diets containing 8, 15, 20, 26 or 36 mg Fe/kg for approximately 10 weeks ad lib.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3032235 TI - Limited proteolysis of type I collagen at hyperreactive sites by class I and II Clostridium histolyticum collagenases: complementary digestion patterns. AB - The initial proteolytic events in the hydrolysis of rat tendon type I collagen by the class I and II collagenases from Clostridium histolyticum have been investigated at 15 degrees C. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis has been used to detect the initial cleavage fragments of both the alpha 1(I) and alpha 2 chains, which migrate at different rates in the buffer system employed. Experiments with the class I collagenases indicate that the first cleavage occurs across all three chains of the triple helix close to the C terminus to produce fragments whose alpha chains have molecular weights of approximately 88,000. The second cleavage occurs near the N-terminus to reduce the molecular weight of the alpha chains to 80,000. Initial proteolysis by the class II collagenases occurs across all three chains at a site in the interior of the collagen triple helix to give N- and C-terminal fragments with alpha-chain molecular weights of 35,000 and 62,000, respectively. The C-terminal fragment is subsequently cleaved to give fragments with alpha-chain molecular weights of 59,000. These results indicate that type I collagen is degraded at several hyperreactive sites by these enzymes. Thus, initial proteolysis by these bacterial collagenases occurs at specific sites, much like the mammalian collagenases. These results with the individual clostridial collagenases provide an explanation for earlier data which indicated that collagen is degraded sequentially from the ends by a crude clostridial collagenase preparation. PMID- 3032236 TI - Laser flash photolysis studies of electron transfer between ferredoxin-NADP+ reductase and several high-potential redox proteins. AB - Complex formation and the kinetics of electron transfer between ferredoxin-NADP+ reductase (FNR) and two structurally homologous acidic 4Fe-4S high-potential ferredoxins (HiPIP's) from Ectothiorhodospira halophila (HP1 and HP2) and two structurally homologous cytochromes c2 from Paracoccus denitrificans and Rhodospirillum rubrum (PC2, and RC2, respectively) have been investigated by gel filtration and laser flash photolysis techniques. Gel filtration studies indicated that complex formation occurred between FNRox and HP1ox or HP2ox at low ionic strength (10 mM) and that the complexes were completely dissociated at high ionic strength (310 mM). Laser flash photolysis using lumiflavin as the reductant demonstrated that both free HP1ox and HP2ox reacted primarily with the anionic form of fully reduced lumiflavin (LFH-), whereas FNR was unreactive. Second-order rate constants of 1 X 10(6) and 0.8 X 10(6) M-1 s-1 were obtained for these reactions at 10 mM ionic strength. Increasing the ionic strength to 310 mM resulted in an approximately 1.5-fold increase in the rate constant. Inclusion of stoichiometric amounts of FNRox into the reaction mixture at low ionic strength led to a 2.5-fold increase in the rate constants. The reaction of 5 deazariboflavin semiquinone (5-dRf.) with the oxidized HiPIP's was also investigated by laser flash photolysis. Second-order rate constants of 3.0 X 10(8) M-1 s-1 (HP1) and 2.5 X 10(8) M-1 s-1 (HP2) were obtained for the free proteins at 10 mM ionic strength. Under the same conditions, 5-dRf. reacted with free FNRox, resulting in the formation of the neutral protein-bound semiquinone (FNR.), with a second-order rate constant of 6 X 10(8) M-1 s-1.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3032237 TI - The Mr 93,000 polypeptide of the postsynaptic glycine receptor complex is a peripheral membrane protein. AB - The glycine receptor of mammalian spinal cord is an oligomeric membrane protein that, after affinity purification on aminostrychnine-agarose or immobilized antibody, contains three polypeptides of Mr 48,000, 58,000, and 93,000. Here, the association and the properties of the polypeptides of the rat glycine receptor were investigated. Upon phase partitioning in the nonionic detergent Triton X 114, the three receptor polypeptides behaved as a hydrophilic protein complex exhibiting phospholipid binding. Sucrose gradient centrifugation or gel filtration in the presence of dithiothreitol and Triton X-100 separated the Mr 93,000 polypeptide from the Mr 48,000 and 58,000 polypeptides, which harbor the antagonist binding site of the glycine receptor. Alkaline or dimethylmaleic acid anhydride treatment of crude synaptic membrane fractions resulted in extraction of the Mr 93,000 polypeptide. Lectin binding was observed for the Mr 48,000 and 58,000 glycine receptor subunits but not the Mr 93,000 polypeptide. These results indicate that the Mr 93,000 polypeptide is a peripheral membrane protein that is located at the cytoplasmic face of the postsynaptic glycine receptor complex. PMID- 3032238 TI - Lateral mobility of reconstituted Sendai virus envelope glycoproteins on human erythrocytes: correlation with cell-cell fusion. AB - We have recently employed fluorescence photobleaching recovery (FPR) to demonstrate that the envelope glycoproteins of Sendai virions become laterally mobile on the surface of human erythrocytes following fusion [Henis, Y. I., Gutman, O., & Loyter, A. (1985) Exp. Cell Res. 160, 514-526]. In order to investigate whether this lateral mobilization is involved in the mechanism of virally mediated cell-cell fusion, or is merely a result of viral envelope-cell fusion, we have now performed FPR studies on erythrocytes fused with reconstituted Sendai virus envelopes (RSVE). These RSVE, which were prepared by solubilization of Sendai virions with Triton X-100 followed by removal of the detergent through adsorption to SM-2 Bio-beads, fused with human erythrocytes as efficiently as native virions but induced cell-cell fusion to a much lower degree. The fraction of the viral envelope glycoproteins that became laterally mobile in the erythrocyte membrane following fusion was markedly lower in the case of RSVE than in the case of native virions. The lower cell-cell fusion activity of the RSVE does not appear to be due to inactivation of the viral fusion protein, since the envelope-cell fusion and hemolytic activities of the RSVE were similar to those of native virions. Moreover, fusion with RSVE or with native virions resulted in the incorporation of rather similar amounts of viral glycoproteins into the cell membrane. Since the reduced fraction of laterally mobile viral glycoproteins correlates with the lower cell-cell fusion activity of the RSVE.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3032239 TI - Reconstitution of membrane proteins: sequential incorporation of integral membrane proteins into preformed lipid bilayers. AB - Several integral membrane proteins can be inserted sequentially into preformed unilamellar vesicles (ULV's) composed of dimyristoylphosphatidylcholine (DMPC) and cholesterol in a gel phase. Thus, proteoliposomes of DMPC, cholesterol, and bacteriorhodopsin from Halobacterium halobium rapidly incorporate UDPglucuronosyltransferase (EC 2.4.1.17) from pig liver microsomes, cytochrome oxidase from beef heart mitochondria, and additional bacteriorhodopsin, added sequentially. This process of spontaneous incorporation can be regulated to produce complex artificial membranes that contain phospholipids and proteins at ratios (mol/mol) equivalent to what is found in biological membranes. The ability of the lipid-protein bilayers to incorporate additional integral membrane proteins is not affected by annealing of the proteoliposomes at 37 degrees C nor by the order of addition of the proteins. Bacteriorhodopsin-containing vesicles formed by the sequential addition of integral membrane proteins demonstrate light driven proton pumping. Therefore, they have retained a vesicular structure. Vesicles containing one or two different proteins will fuse with each other at 21 degrees C or with ULV's devoid of proteins. Incorporation of bacteriorhodopsin or UDPglucuronosyltransferase into proteoliposomes containing DMPC, with or without cholesterol as impurity, also occurs above the phase transition for DMPC. The presence of a protein in a liquid-crystalline bilayer provides the necessary condition for promoting the spontaneous incorporation of other membrane proteins into preformed bilayers. PMID- 3032240 TI - Specific activation of transcription initiation by the sequence-specific DNA binding agents distamycin A and netropsin. AB - A series of promoters with nine base-pair substitutions in the spacer DNA separating the -10 and -35 regions was used to demonstrate that Escherichia coli RNA polymerase is sensitive to events affecting the spacer DNA--a region not directly contacted by the enzyme. The drugs distamycin A and netropsin specifically enhanced the rate of functional complex formation at a promoter bearing a substitution of nonalternating A-T base pairs. The effect is exerted at an early step in the RNA polymerase-promoter interaction. We hypothesize that a drug-induced structural alteration in the spacer DNA occurs, similar to that normally resulting from RNA polymerase binding. These findings are relevant to an understanding of potential mechanisms of transcription activation. PMID- 3032241 TI - Mammalian alpha-polymerase: cloning of partial complementary DNA and immunobinding of catalytic subunit in crude homogenate protein blots. AB - A new polyclonal antibody against the alpha-polymerase catalytic polypeptide was prepared by using homogeneous HeLa cell alpha-polymerase. The antibody neutralized alpha-polymerase activity and was strong and specific for the alpha polymerase catalytic polypeptide (Mr 183,000) in Western blot analysis of crude extracts of HeLa cells. The antibody was used to screen a cDNA library of newborn rat brain poly(A+) RNA in lambda gt11. A positive phage was identified and plaque purified. This phage, designated lambda pol alpha 1.2, also was found to be positive with an antibody against Drosophila alpha-polymerase. The insert in lambda pol alpha 1.2 (1183 base pairs) contained a poly(A) sequence at the 3' terminus and a short in-phase open reading frame at the 5' terminus. A synthetic oligopeptide (eight amino acids) corresponding to the open reading frame was used to raise antiserum in rabbits. Antibody affinity purified from this serum was found to be immunoreactive against purified alpha-polymerase by enzyme-linked immunosorbent assay and was capable of immunoprecipitating alpha-polymerase. This indicated the lambda pol alpha 1.2 insert encoded an alpha-polymerase epitope and suggested that the cDNA corresponded to an alpha-polymerase mRNA. This was confirmed in hybrid selection experiments using pUC9 containing the cDNA insert and poly(A+) RNA from newborn rat brain; the insert hybridized to mRNA capable of encoding alpha-polymerase catalytic polypeptides. Northern blot analysis of rat brain poly(A+) RNA revealed that this mRNA is approximately 5.4 kilobases. PMID- 3032242 TI - Allosteric interactions in sipunculid and brachiopod hemerythrins. AB - Chemical and spectroscopic consequences of allosteric interactions for ligand binding to sipunculid (Phascolopsis gouldii) and brachiopod (Lingula reevii) hemerythrins (Hrs) have been investigated. Possible allosteric effectors for homotropic effects in sipunculid Hrs have been examined, but only reduction in ligand affinity is observed without cooperativity. In contrast to sipunculid Hr, L. reevii Hr binds O2 cooperatively in the pH range 7-8 and exhibits a Bohr effect. Spectroscopic comparisons of the sipunculid and brachiopod Hrs show no significant differences in the active site structures; therefore, modulation of oxygen affinity is attributable to effects linking the site to quaternary structural changes in the octamer. Oxygen equilibria can be fit with a conformational model incorporating a minimum of three states, tensed (T), relaxed (R), and an R-T hybrid. Resonance Raman spectra of L. reevii oxyHr show a shift in the peroxo stretching frequency when the pH is lowered from pH 7.7 (predominantly R oxyHr) to pH 6.3 (a mixture of R, T, and R-T hybrid), but P. gouldii Hr does not have a frequency shift under the same conditions. In contrast to hemoglobins, ligand binding to the deoxy and met forms is noncooperative for brachiopod (and sipunculid) Hrs. It is thus suggested that conformational changes in the protein are linked to the oxidation state change that accompanies oxygenation of the coupled binuclear iron site (deoxy [FeIIFeII]----oxy [FeIIIFeIII]). The total allosteric energy expended in oxygenation is about 1.4 kcal/mol, and such a shift is possible in the relaxed-tense conversion with relatively limited constraints of the iron coordination environment via the protein quaternary structure. The mechanism of cooperativity in the binuclear copper oxygen carrier hemocyanin is discussed in light of these results. PMID- 3032243 TI - The majority of cellular fatty acid acylated proteins are localized to the cytoplasmic surface of the plasma membrane. AB - The BC3Hl muscle cell line was previously reported to contain a broad array of fatty acid acylated proteins [Olson, E. N., Towler, D. A., & Glaser, L. (1985) J. Biol. Chem. 260, 3784-3790]. Palmitate was shown to be attached to membrane proteins posttranslationally through thiol ester linkages, whereas myristate was attached cotranslationally, or within seconds thereafter, to soluble and membrane bound proteins through amide linkages [Olson, E. N., & Spizz, G. (1986) J. Biol. Chem. 261, 2458-2466]. The temporal and subcellular differences between palmitate and myristate acylation suggested that these two classes of acyl proteins might follow different intracellular pathways to distinct subcellular membrane systems or organelles. In this study, we examined the subcellular localization of the major fatty acylated proteins in BC3Hl cells. Palmitate-containing proteins were localized to the plasma membrane, but only a subset of myristate-containing proteins was localized to this membrane fraction. The majority of acyl proteins were nonglycosylated and resistant to digestion with extracellular proteases, suggesting that they were not exposed to the external surface of the plasma membrane. Many proteins were, however, digested during incubation of isolated membranes with proteases, which indicates that these proteins face the cytoplasm. Two-dimensional gel electrophoresis of proteins labeled with [3H]palmitate and [3H]myristate revealed that individual proteins were modified by only one of the two fatty acids and did not undergo both N-linked myristylation and ester-linked palmitylation. Together, these results suggest that the majority of cellular acyl proteins are routed to the cytoplasmic surface of the plasma membrane, and they raise the possibility that fatty acid acylation may play a role in intracellular sorting of nontransmembranous, nonglycosylated membrane proteins. PMID- 3032244 TI - Cloning and sequence determination of a complementary DNA related to human liver microsomal cytochrome P-450 S-mephenytoin 4-hydroxylase. AB - A cDNA sequence related to the human cytochrome P-450 responsible for S mephenytoin 4-hydroxylation (P-450MP) has been isolated from a human liver bacteriophage lambda gt11 library with antibodies specific for P-450MP. The total length of the cDNA is 2.5 kilobases (kb), of which there is a 1.6-kb EcoRI fragment coding for all but five amino acids corresponding to the N-terminus of the protein and including a small noncoding region at the 3' end. This 1.6-kb fragment has been sequenced and used as a probe to analyze human genomic DNA and liver RNA. The sequence shows extensive sequence similarity with that of rabbit liver cytochrome P-450 progesterone 21-hydroxylase [Tukey, R. H., Okino, S., Barnes, H., Griffin, K. J., & Johnson, E. F. (1985) J. Biol. Chem. 260, 13347 13354], and this cDNA, like the rabbit clone, appears to be part of a multigene family. At least two liver mRNA species, 2.2 kb and 3.5 kb, hybridize to the cDNA sequence. The cloning of this gene should aid in analyzing the molecular basis for the genetic polymorphism of S-mephenytoin 4-hydroxylation reported in humans. PMID- 3032245 TI - Head group and chain length dependence of phospholipid self-assembly studied by spin-label electron spin resonance. AB - The critical micelle concentrations (cmc's) of a variety of spin-labeled phospholipids, 1-acyl-2-[4-(4,4-dimethyloxazolidine-N-oxyl)valeryl]-sn-glycero-3 pho sph o derivatives, have been determined by electron spin resonance (ESR) spectroscopy. The narrow, three-line ESR spectra of the rapidly tumbling monomers are clearly distinguished from the spin-spin broadened spectra of the micellar aggregates, allowing a direct determination of the concentrations of the two species. The influence of both the hydrocarbon chain length and the polar head group on the energetics of self-assembly has been studied. For phosphatidylcholine, 1n [cmc] decreases linearly with the length of the sn-1 chain. The gradient of this linear dependence corresponds to a free energy of transfer of the monomer from the aqueous phase to the micelle of delta Gtr = 1.1RT per CH2 group. The cmc's of the 1-lauroyl derivatives of both phosphatidylcholine and phosphatidylglycerol have relatively shallow, biphasic temperature dependences with a minimum at approximately 20 degrees C. Both of these properties are characteristic of the hydrophobic effect, with the free energy of transfer being slightly less than that for the solubility of n hydrocarbons in water, corresponding to the reduced configurational entropy of the lipid chains in the micellar state. The cmc's of the 1-lauroyl derivatives of the phospholipids in 0.15 M NaCl, for their various charge states, are as follows: phosphatidic acid(2-), 0.77 mM; phosphatidic acid(1-), 0.13 mM; phosphatidylserine(1-), 0.24 mM; phosphatidylglycerol(1-), 0.17 mM; phosphatidylcholine, 0.10 mM; phosphatidylethanolamine, 0.05 mM.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3032246 TI - 2,3-diphosphoglycerate phosphatase activity of phosphoglycerate mutase: stimulation by vanadate and phosphate. AB - The binding of inorganic vanadate (Vi) to rabbit muscle phosphoglycerate mutase (PGM), studied by using 51V nuclear magnetic resonance spectroscopy, shows a sigmoidal dependence on vanadate concentration with a stoichiometry of four vanadium atoms per PGM molecule at saturating [Vi]. The data are consistent with binding of one divanadate ion to each of the two subunits of PGM in a noncooperative manner with an intrinsic dissociation constant of 4 X 10(-6) M. The relevance of this result to other studies which have shown that the Vi stimulated 2,3-diphosphoglycerate (2,3-DPG) phosphatase activity of PGM has a sigmoidal dependence on [Vi] with a Hill coefficient of 2.0 is discussed. At pH 7.0, inorganic phosphate has little effect on the 2,3-DPG phosphatase activity of PGM, even at concentrations as high as 50 mM. Similarly, 25 microM Vi has little effect on the phosphatase activity. However, in the presence of 25 microM Vi, a phosphate concentration of 20 mM increases the phosphatase activity by more than 3-fold. This behavior is rationalized in terms of activation of the phosphatase activity by a phosphate/vanadate mixed anhydride. This interpretation is supported by the observation of strong activation of the phosphatase activity by inorganic pyrophosphate. A molecular mechanism for the observed effects of vanadate is proposed, and the relevance of this study to the possible use of vanadate as a therapeutic agent for the treatment of sickle cell anemia is discussed. PMID- 3032247 TI - Studies of the active site of cytochrome P-450scc with a high-affinity spin labeled inhibitor. AB - The intramolecular site of P-450scc for conversion of cholesterol to pregnenolone involves a substrate site, an active site, and a site for transmission of electrons. The substrate site was studied with a high-affinity, high-potency nitroxide spin-labeled inhibitor of cholesterol side-chain cleavage. This substance, 17 alpha-hydroxy-11-deoxycorticosterone nitroxide (SL-V), has an affinity comparable to that of the most active substrate inhibitors ever reported and 2-50 times greater than that of the natural substrate cholesterol. Competition experiments with cholesterol and its analogues confirmed that SL-V binds reversibly to the substrate site. Titration experiments showed a single binding site on the P-450 molecule. The substrate site is on the apoprotein and has little or no direct interaction with the heme. Spin-spin interactions between the Fe3+ and side-chain or A-ring spin-labeled groups could not be demonstrated, which is consistent with carbons 22 and 20 being closest to the heme iron. We postulate that substrate disrupts a histidine nitrogen coordination with the heme iron and induces conformational changes in the apoprotein. These changes lead to increased affinity for iron-sulfur protein. PMID- 3032248 TI - Detergents as probes of reconstituted rat liver cytochrome P-450 function. AB - A series of 16 ionic, zwitterionic, and nonionic detergents have been used to perturb the catalytic activities of major cytochrome P-450 (P-450) forms from untreated (UT-A), phenobarbital-treated (PB-B) and beta-naphthoflavone-treated (BNF-B) rats in reconstituted systems with NADPH--P-450 reductase Detergent effects on R warfarin hydroxylase activities were correlated with detergent effects on the quaternary structures of P-450 and reductase, and on their 1:1 complexes as determined by gel exclusion chromatography using sodium cholate as a prototype detergent. The detergent concentrations used did not in most cases affect rates of NADPH-dependent reduction of cytochrome c by the reductase. With P-450 BNF-B, ionic and zwitterionic detergents enhanced warfarin hydroxylase activities at low concentrations and produced marked inhibition at higher concentrations, while nonionic detergents only inhibited. With P-450 UT-A, some nonionic and zwitterionic detergents increased rates at low concentrations and inhibited at higher concentrations. P-450 PB-B was inhibited by detergents of all three classes at low and high concentrations. The concentrations of a detergent required to affect 50% inhibition differed for the three P-450s, suggesting, together with the differential susceptibilities to detergent-mediated rate enhancing effects, that the reductase interacts functionally differently with the three P-450s. Chromatographic studies demonstrated that concentrations of sodium cholate which optimally enhanced metabolic rates with P-450 BNF-B facilitated the uptake of the P-450 into the functional reductase/P-450 complex, and higher concentrations of cholate, which completely inhibited activity, produced profound disruptions of the complex. The data have provided insight into the functional interactions required for monooxygenase activity. PMID- 3032249 TI - Observation of a chloride-dependent intermediate during catalysis by angiotensin converting enzyme using radiationless energy transfer. AB - Stopped-flow radiationless energy-transfer kinetics have been used to examine the effects of chloride on the hydrolysis of Dns-Lys-Phe-Ala-Arg by angiotensin converting enzyme. The kinetic constants for hydrolysis at pH 7.5 and 22 degrees C in the presence of 300 mM sodium chloride were KM = 28 microM and kcat = 110 s 1, and in its absence, KM = 240 microM and kcat = 68 s-1. The apparent binding constant for chloride was 4 mM, and the extent of chloride activation in terms of kcat/KM was 14-fold. The effects of chloride on the pre-steady-state were examined at 2 degrees C. In the presence of chloride, two distinct enzyme substrate complexes were observed, suggesting multiple steps in substrate binding. The initial complex was formed during the mixing period (kobsd greater than 200 s-1) while the second complex was formed much more slowly (kobsd = 40 s 1 when [S] = 5 microM and [NaCl] = 150 mM). Strikingly, in the absence of chloride, only a single, rapidly formed enzyme-substrate complex was observed. These results are consistent with a nonessential activator kinetic mechanism in which the slow step reflects conversion of an initially formed complex, (E X Cl- X S)1, to a more tightly bound complex, (E X Cl- X S)2. PMID- 3032250 TI - Catalysis by human leukocyte elastase: proton inventory as a mechanistic probe. AB - Proton inventories (rate measurements in mixtures of H2O and D2O) were determined for the human leukocyte elastase catalyzed hydrolyses of thiobenzyl esters and p nitroanilides of the peptides MeOSuc-Val, MeOSuc-Alan-Pro-Val (n = 0-2), and MeOSuc-Alan-Pro-Ala (n = 1 or 2). The dependencies of k2/Ks on mole fraction of solvent deuterium for the p-nitroanilides are "dome-shaped" and were fit to a model that incorporates the mechanistic features of generalized solvent reorganization when substrate binds to enzyme and partial rate limitation of k2/Ks by physical and chemical steps [Stein, R. L. (1985) J. Am. Chem. Soc. 107, 7768-7769]. The proton inventories for the deacylation of MeOSuc-Val-HLE and MeOSuc-Pro-Val-HLE are linear while those for the deacylation of MeOSuc-Ala-Pro Val-HLE and MeOSuc-Ala-Ala-Pro-Val-HLE are "bowl-shaped" and could be fit to a quadratic dependence of rate on mole fraction of deuterium. These results are interpreted to suggest that the correct operation of the catalytic triad is dependent on substrate structure. Minimal substrates, which cannot interact with elastase at remote subsites, are hydrolyzed via a mechanism involving simple general-base catalysis by the active site histidine and transfer of a single proton in the rate-limiting transition state. In contrast, tri- and tetrapeptide substrates, which are able to interact at remote subsites, are hydrolyzed by a more complex mechanism of protolytic catalysis involving full functioning of the catalytic triad and transfer of two protons in the rate-limiting transition state. Finally, the proton inventories for the deacylation of MeOSuc-Ala-Pro-Ala HLE and MeOSuc-Ala-Ala-Pro-Ala-HLE are dome-shaped and suggest that the chemical events of acyl-enzyme hydrolysis are only partially rate limiting for these reactions and that some other physical step is also partially rate limiting. PMID- 3032253 TI - Electron-spin resonance studies of the bound iron-sulfur centers in Photosystem I. II. Correlation of P-700 triplet production with urea/ferricyanide inactivation of the iron-sulfur clusters. AB - Photosystem I charge separation in a subchloroplast particle isolated from spinach was investigated by electron spin resonance (ESR) spectroscopy following graduated inactivation of the bound iron-sulfur centers by urea-ferricyanide treatment. Previous work demonstrated a differential decrease in iron-sulfur centers A, B and X which indicated that center X serves as a branch point for parallel electron flow through centers A and B (Golbeck, J.H. and Warden, J.T. (1982) Biochim. Biophys. Acta 681, 77-84). We now show that during inactivation the disappearance of iron-sulfur centers A, B, and X correlates with the appearance of a spin-polarized triplet ESR signal with [D] = 279 X 10(-4) cm-1 and [E] = 39 X 10(-4) cm-1. The triplet resonances titrate with a midpoint potential of +380 +/- 10 mV. Illumination of the inactivated particles results in the generation of an asymmetric ESR signal with g = 2.0031 and delta Hpp = 1.0 mT. Deconvolution of the P-700+ contribution to this composite resonance reveals the spectrum of the putative primary acceptor species A0, which is characterized by g = 2.0033 +/- 0.0004 and delta Hpp = 1.0 +/- 0.2 mT. The data presented in this report do not substantiate the participation of the electron acceptor A1 in PS I electron transport, following destruction of the iron-sulfur cluster corresponding to center X. We suggest that A1 is closely associated with center X and that this component is decoupled from the electron-transport path upon destruction of center X. The inability to photoreduce A1 in reaction centers lacking a functional center X may result from alteration of the reaction center tertiary structure by the urea-ferricyanide treatment or from displacement of A1 from its binding site. PMID- 3032252 TI - Purification of highly active cytochrome bc1 complexes from phylogenetically diverse species by a single chromatographic procedure. AB - A method has been developed for purification of highly active ubiquinol cytochrome c oxidoreductase (cytochrome bc1) complexes from wild-type Rhodobacter sphaeroides, Rhodobacter capsulatus MT1131, bovine heart and yeast mitochondria. This is the first report of the isolation of cytochrome bc1 complex from a wild type strain of Rb. sphaeroides and from any strain of Rb. capsulatus. The purification involves extraction of membranes with dodecyl maltoside and two successive DEAE column chromatography steps. All of the resulting bc1 complexes are free of succinate dehydrogenase and cytochrome c oxidase activities. The purified bc1 complexes from both photosynthetic bacteria contain four polypeptide subunits, although the molecular weights of some of their subunits differ. They are also free of reaction center and light-harvesting pigments and polypeptides. The turnover number of the Rb. sphaeroides complex is 128 s-1, and that of the Rb. capsulatus complex is 64 s-1. The bc1 complex from bovine heart contains eight polypeptides and has a turnover number of 1152 s-1, while the yeast complex contains nine polypeptides and has a turnover number of 219 s-1. The activities of these complexes are equal to or better than those commonly obtained by previously reported methods. This method of purification is relatively simple, reproducible, and yields cytochrome bc1 complexes which largely retain the turnover number of the starting material and are pure on the basis of optical spectra, enzymatic activities and polypeptide composition. The purification of cytochrome bc1 complexes from energy-transducing membranes which differ markedly in their lipid and protein composition makes it likely that with minor modifications this method could be applied to species other than those described here. PMID- 3032251 TI - Functional differences in the beta gamma complexes of transducin and the inhibitory guanine nucleotide regulatory protein. AB - We have examined the mechanism of inhibition of adenylate cyclase using the purified alpha and beta gamma subunits of bovine brain inhibitory guanine nucleotide regulatory protein (Ni) (i.e., alpha i and beta gamma N) and bovine retinal transducin (alpha T and beta gamma T) in reconstituted phospholipid vesicle systems. The addition of beta gamma N or beta gamma T to lipid vesicles containing the pure stimulatory guanine nucleotide regulatory protein (Ns) from human erythrocytes as well as a resolved preparation of the catalytic moiety (C) of bovine caudate adenylate cyclase results in significant inhibition of guanine nucleotide stimulated cyclase activity (80-90%). The inhibition by these beta gamma subunit complexes appears to fully account for the inhibitory effects observed with holo-Ni or holotransducin. A variety of structure-function comparisons of the beta gamma N and beta gamma T complexes were performed in order to further probe the molecular mechanisms involved in the inhibitory pathway. Whereas the beta subunits of beta gamma N and beta gamma T appear to be very similar, if not identical, on the basis of comparisons of their gel electrophoretic mobility and immunological cross-reactivity, clear differences exist in the apparent structures of gamma N and gamma T. Interestingly, functional differences are observed in the effectiveness of these two beta gamma complexes to inhibit adenylate cyclase activity. Specifically, while both beta gamma N and beta gamma T are capable of effecting significant levels of inhibition of the guanine nucleotide stimulated activities, the beta gamma N complex is consistently more potent than beta gamma T in inhibiting these activities.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3032254 TI - Inhibition of transepithelial osmotic water flow by blockers of the glucose transporter. AB - On the basis of evidence derived mostly from human erythrocytes, it has been suggested that water traverses cell membranes through membrane-spanning proteins such as the anion channel or the glucose transporter acting as water pores. However, specific inhibitors of such permeation processes have not been found to block water transport, and hence a precise identification of the water route has not been possible so far. We have investigated this issue by characterizing the osmotic flows across a fluid-transporting epithelium, the rabbit corneal endothelium. The rate of such flows was monitored continuously as a function of time. We confirmed prior findings of an inhibition by PCMBS on osmotic water flow, and lack of inhibition by DTNB and DIDS. On the other hand, we have found for the first time that several blockers of glucose facilitated diffusion, namely, phloretin (2 mM), phloridzin (2 mM), diallyldiethylstilbestrol (0.1 mM), cytochalasin B (20 micrograms/ml), and ethylidene-D-glucose (200 mM), all clearly inhibit osmotic flow. Our evidence is consistent with the hypothesis that both water and glucose may traverse these cell membranes through the same channel-like pathway contained in the glucose transporter membrane-spanning protein. PMID- 3032255 TI - The localization of cholinephosphotransferase in the outer membrane of guinea-pig lung mitochondria. AB - The activity of cholinephosphotransferase was measured in the subcellular fractions of guinea-pig lung. The specific activity of the enzyme was highest in a fraction, intermediate in density between mitochondria and microsomes. Similar subcellular distribution patterns were observed for both cholinephosphotransferase and rotenone-insensitive NADH-cytochrome c reductase, an enzyme associated with the outer membrane of mitochondria and endoplasmic reticulum, suggesting that cholinephosphotransferase may be localized in both of these organelles. The distribution of cholinephosphotransferase activity in the subfractions of mitochondria and the intermediate fractions recovered by linear density gradient paralleled that of the mitochondrial outer membrane marker enzyme, monoamine oxidase. RNA content of a subfraction enriched in cholinephosphotransferase and monoamine oxidase was not typical to that of either rough or smooth endoplasmic reticulum. The results of this study suggest that in guinea-pig lung, cholinephosphotransferase is localized in both the outer membrane of mitochondria, and the endoplasmic reticulum. PMID- 3032256 TI - Quinidine and melittin both decrease the fluidity of liver plasma membranes and both inhibit hormone-stimulated adenylate cyclase activity. AB - Increasing concentrations of either quinidine or melittin gave a dose-dependent inhibition of both the glucagon- and fluoride-stimulated activities of adenylate cyclase in the liver plasma membranes. At similar concentrations these agents increased the order of liver plasma membranes as detected by a fatty acid ESR probe, doxyl stearic acid. This increase in bilayer order (decrease in 'fluidity') is suggested to explain the inhibitory action of quinidine on adenylate cyclase activity but only in part contributes to the inhibitory action of melittin on adenylate cyclase. Arrhenius plots of fluoride-stimulated activity became non-linear in the presence of either quinidine or melittin, with a single well-defined break occurring at around 12 degrees C in each instance. Arrhenius plots of the glucagon-stimulated activity also exhibited such a novel break at around 12 degrees C when either quinidine or melittin were present as well as exhibiting a break at around 28 degrees C, as was seen in the absence of these ligands. The fatty acid spin probe inserted into liver plasma membranes detected a novel lipid phase separation occurring at around 12 degrees C when either quinidine or melittin was present and showed that the lipid phase separation occurring at around 28 degrees C in native membranes was apparently unaffected by these ligands. PMID- 3032258 TI - Inositol lipids and cell proliferation. PMID- 3032257 TI - The spontaneous incorporation of proteins into preformed bilayers. PMID- 3032259 TI - Mechanisms of retrovirus-induced leukaemia: selected aspects. PMID- 3032260 TI - Expression of human antithrombin III in Saccharomyces cerevisiae and Schizosaccharomyces pombe. AB - Recombinant plasmids were constructed that direct the synthesis of human antithrombin III in baker's yeast, Saccharomyces cerevisiae, and the fission yeast, Schizosaccharomyces pombe. The signal sequence of antithrombin III was recognized by both yeast species, and antithrombin III was secreted into the medium. When the signal sequence was replaced by a sequence of ten arbitrary amino acids, the product expressed from such a construct stayed inside the cell. Antithrombin III was glycosylated by the baker's and fission yeast and was immunologically identical to antithrombin III isolated from human plasma. Antithrombin III isolated from the culture media of recombinant yeasts was biologically active, as could be shown by progressive inhibitor activity and heparin cofactor activity. PMID- 3032262 TI - Nuclease S1-sensitive sites on superhelical DNA molecules carrying the LTR region of Moloney murine leukemia virus. AB - The long terminal repeat (LTR) from proviral DNA of Moloney murine leukemia virus (Mo-MLV) was cloned on a derivative of pBR322, and after introducing superhelical torsions into the resulting recombinant, the sites of conformational transition were investigated by the nuclease S1-digestion method. With an increase in the negative linking differences, fourteen dominant cutting sites were identified, of which two were mapped inside the LTR and one at the 3' end of the LTR. By searching the sequence data, all these sites were localized in the regions having either palindromic sequences or AT-rich sequences. Free energy calculation for the local secondary structure on one strand indicated that nuclease S1 attacked the palindromic sequence regions which could form relatively stable hairpin structures. Under the conditions used, no correlation was found between the S1 sensitive sites and the potential Z-DNA-forming regions, including those within the enhancer sequence. PMID- 3032261 TI - Nuclear non-polyadenylated RNAs containing the first intervening sequence of the major late premessenger RNA from adenovirus-2: characterization and distribution in ribonucleoproteins. AB - The nuclear non-polyadenylated RNA from HeLa cells infected with adenovirus-2 was examined for the presence of molecules containing the first intervening sequence (IVS1) of the major late premessenger RNA. Four molecules with the approximate size of free IVS1 in sucrose gradients (1021 nucleotides) were separated by polyacrylamide gel electrophoresis and characterized by complementary methods: S1 nuclease mapping, susceptibility to debranching enzyme, RNAase-H-directed cleavage. The results indicate that the most abundant RNA form is the excised lariat IVS1. We also find linear IVS1 and a randomly nicked lariat, the latter probably being made during RNA isolation. The fourth RNA is a leader 1-IVS1 molecule. No truncated IVS1 which might indicate that IVS1 is excised by several cycle of cleavage-ligation was detected. A study of the distribution of the four RNAs in hnRNP shows that they are part of RNPs of about 70 S. However, each RNP has distinct sedimentation characteristics and sensitivity to salt dissociation. Together, the results suggest that the excised lariat IVS1 is released from the large late premRNP under the form of a 70 S RNP, where it is linearized. PMID- 3032263 TI - Electronic absorption and EPR spectroscopy of copper alcohol dehydrogenase: pink, violet and green forms of a type 1 copper center analog. AB - Blue and non-blue states of the copper center in copper-substituted alcohol dehydrogenase (EC 1.1.1.1) can be attained by coenzyme binding and/or ligand binding to the copper ion. Copper alcohol dehydrogenase has been studied by electronic absorption, CD and EPR spectroscopy in the presence and absence of coenzyme. On the basis of previous work on blue (Type 1) copper proteins with a CuSS*N2 chromophore the assignment of charge transfer transitions in copper alcohol dehydrogenase is discussed. The latter contains a CuS2N(OH2) unit in the ligand-free protein and a CuS2N2 unit in the ternary complex with NAD+ and pyrazole. It is proposed that the energy of the charge transfer transitions can be used as a structural marker in combination with EPR data. A comparison is made between the spectroscopic properties of the ternary complex of copper alcohol dehydrogenase and the copper centers in stellacyanin and cytochrome-c oxidase (CuA) in order to test the validity of recent structural models of the type CuS2N2, i.e., a cupric ion coordinated to two thiolate ligands. Finally, a close resemblance between the electronic absorption spectra of copper alcohol dehydrogenase and those of other variants of Type 1 copper centers such as the 'unusual' copper center of nitrous oxide reductase is noted as an indication of similar coordination environments. PMID- 3032264 TI - Buffer, pH, and ionic strength effects on the (Na+ + K+)-ATPase. AB - A dog kidney (Na+ + K+)-ATPase preparation also catalyzes K+-independent and K+ activated phosphatase reactions with p-nitrophenyl phosphate as substrate. K+ independent activity increases with declining pH over the range 7.5 to 5.8, whereas the other two activities decrease. The increased K+-independent activity is similar with imidazole, histidine, and several Good buffers, and is thus attributable to free H+, probably by affecting enzyme conformations rather than by changing affinity for Mg2+ or substrate or by H+ occupying specific K+-sites. The decrease in K+-phosphatase and (Na+ + K+)-ATPase activities with pH also occurs similarly with those buffers, and is not due to changes in apparent affinity for substrate or for cation activators. However, the Good buffers Pipes and ADA inhibit the K+-independent phosphatase reaction strongly, the K+ activated reaction moderately, and the (Na+ + K+)-ATPase reaction little; both contain two acidic groups, unlike the other buffers tested. Inhibition of the phosphatase reaction by Pipes is associated with a decreased apparent affinity for K+ and an increased sensitivity to inhibition by Na+ and ADP, consistent with Pipes hindering conformational transitions to the E2 enzyme forms required for phosphatase hydrolytic activity. PMID- 3032265 TI - Formation of 6,15-diketoprostaglandin F1 alpha from prostaglandin G2 by bovine aortic endothelial cells. AB - Besides 6-ketoprostaglandin F1 alpha, bovine aortic endothelial cells also produced considerable amounts of 6,15-diketoprostaglandin F1 alpha from arachidonic acid, either exogenously added or released from cellular phospholipids. Incubations of particulate fractions of endothelial cells with the cyclic endoperoxides prostaglandin G2 and prostaglandin H2 showed that 6,15 diketoprostaglandin F1 alpha is formed by the action of prostaglandin I2 synthetase on prostaglandin G2. The labile metabolite 15-hydroperoxyprostaglandin I2 is then converted nonenzymatically to the 15-keto derivative. In the presence of reduced glutathione, quantitative analysis of both metabolites by gas chromatography-mass spectrometry showed a significant decrease of 6,15 diketoprostaglandin F1 alpha formation, whereas prostaglandin I2 synthesis was markedly increased. This shift seems to be due to a stimulation of peroxidase by GSH, a well known cofactor of this enzyme. Thus, it seems that a decreased endothelial prostaglandin I2 formation may occur when cellular glutathione levels are reduced as a consequence of oxidant injury and lipid peroxidation. Additionally, ferrous ions seems to be involved in the regulation of endothelial prostaglandin I2 synthesis, since Desferal, a specific ferrous ion chelator that might have antimetastatic properties, produced a pronounced shift from 6,15 diketoprostaglandin F1 alpha to the 6-keto derivative, i.e., prostaglandin I2. PMID- 3032266 TI - Inhibition of leukotriene formation in human leukocytes by halothane. AB - The effects of an inhalation anesthetic, halothane (2-bromo-2-chloro-1,1,1 trifluoroethane) on the formation of 5-lipoxygenase metabolites such as leukotriene B4, 5(S)-hydroxyeicosatetraenoic acid (5-HETE), 6-trans-isomers of leukotriene B4 and leukotriene C4 were studied in human leukocytes stimulated with calcium ionophore A23187. Halothane inhibited the formation of all these metabolites dose dependently and the formation was restored by removal of the drug. The anesthetic also reversibly inhibited the release of [3H]arachidonic acid from neutrophils with a half-inhibition concentration of less than 0.19 mM. The formation of 5-lipoxygenase metabolites was not inhibited by the anesthetic when leukocytes were stimulated with the ionophore in the presence of exogenous arachidonic acid. These observations indicate that the inhibitory effect of halothane on the formation of 5-lipoxygenase metabolites in leukocytes is mainly due to the inhibition of arachidonic acid release. PMID- 3032267 TI - Enhanced release and synthesis of lipoprotein lipase in rat heart cell cultures exposed to high concentrations of Hepes. AB - While attempting to optimize conditions for synthesis of lipoprotein lipase by cultured heart cells, we encountered an unexpected rise in enzyme activity when media were supplemented inadvertently with 100 mM Hepes buffer (4-(2 hydroxyethyl)-1-piperazineethanesulphonic acid). This finding was further investigated and optimal results were obtained at pH 7.0-7.2. The increase in lipoprotein lipase activity was time dependent; after 3-6 h there was a rise in medium activity but cellular activity increased only after 24 h. The increased enzyme activity was defined as lipoprotein lipase by inhibition with antiserum to rat adipose tissue lipoprotein lipase. A 72-h exposure to Hepes resulted in a 30% increase in the incorporation of [35S]methionine into cellular proteins and a 2 fold increase into heparin-releasable proteins. Using heparin Sepharose chromatography and stepwise elution, a lipoprotein lipase enriched fraction was recovered with 2 M NaCl. The amount of [35S]methionine and [3H]galactose incorporated into protein of this fraction derived from Hepes-treated cells was 2 6-fold that of controls. A 4-fold increase in cellular lipoprotein lipase mass in Hepes-treated cells was shown by immunoblotting. Results obtained with Hepes conditioned medium suggest the presence of cell-derived compounds that enhance release and subsequent synthesis of lipoprotein lipase. The effect of Hepes conditioned medium on lipoprotein lipase resembled to some extent that of the addition of heparin. Therefore, it appears that when Hepes is first added to the culture medium, it might promote a release of heparan sulfate or related compounds, possibly by virtue of its negatively charged sulfonic acid residue. The accumulated heparan sulfate could then promote a sustained release of lipoprotein lipase into the culture medium which in turn leads to increased enzyme synthesis. PMID- 3032268 TI - Translocation of CTP: phosphocholine cytidylyltransferase from cytosol to membranes in HeLa cells: stimulation by fatty acid, fatty alcohol, mono- and diacylglycerol. AB - Addition of oleate, oleyl alcohol, or palmitate to HeLa cell medium resulted in a rapid stimulation of PC synthesis and activation of CTP: phosphocholine cytidylyltransferase. Stimulation was optimal with 0.35 mM oleate, 0.3 mM oleyl alcohol and 5 mM palmitate, or 1 mM palmitate if EGTA were added to the medium. The cytidylyltransferase was activated by translocation of the inactive cytosolic form to membranes. In untreated cells approx. 30% of the total cytidylyltransferase was membrane bound, while in treated cells, 80-90% was membrane associated. Addition of bovine serum albumin (10 mg/ml) to cells previously treated with oleate (0.35 mM) rapidly removed cellular fatty acid, and the membrane-bound cytidylyltransferase activity returned to approx. 30%. Similar results were obtained by extraction of membranes with albumin in vitro. Although 95% of the free fatty acid was extracted, 30-40% of the membrane cytidylyltransferase remained bound. Translocation of cytidylyltransferase between isolated cytosol and microsomal fractions was promoted by addition of oleate, palmitate, oleyl alcohol, and monoolein. Addition of diacylglycerol, lysophosphatidylcholine, lysophosphatidylethanolamine, calcium palmitate, and detergents such as Triton X-100, cholate or Zwittergent did not stimulate translocation of the enzyme. Addition of oleoyl-CoA promoited translocation, however, 40% of it was hydrolyzed releasing free oleic acid. Cytosolic cytidylyltransferase bound to microsomes pre-treated with phospholipase C, which had 7-fold elevated diacylglycerol content. Fatty acid-promoted translocation was blocked by Triton X-100, but not by 1 M KCl. These results suggest that a variety of compounds with differing head group size and charge, and number of hydrocarbon chains can function as translocators, and that hydrophobic rather than ionic interactions mediate the binding of cytidylyltransferase to membranes. PMID- 3032269 TI - Binding of CTP: phosphocholine cytidylyltransferase to large unilamellar vesicles. AB - We have studied the binding of CTP: phosphocholine cytidylyltransferase from HeLa cell cytosol to large unilamellar vesicles of egg phosphatidylcholine (PC) or HeLa cell phospholipids that contain various amounts of oleic acid. A fatty acid/phospholipid molar ratio exceeding 10% was required for CTP: phosphocholine cytidylyltransferase binding to liposomes. At a fatty acid/phospholipid molar ratio of 1; 85% of the cytosolic CTP: phosphocholine cytidylyltransferase was bound. The enzyme also bound to liposomes with at least 20 mol% palmitic acid, monoolein, diolein or oleoylacetylglycerol. Oleoyl-CoA did not promote enzyme binding to liposomes. Binding to oleate-PC vesicles was blocked by Triton X-100 but not by 1 M KCl, and was reversed by incubation of the vesicles with bovine serum albumin. Cytidylyltransferase bound to egg PC vesicles that contained 33 mol% oleic acid equally well at 4 degrees C and 37 degrees C. The enzyme also bound to dimyristoyl- and dipalmitoylphosphatidylcholine vesicles containing oleic acid at temperatures below the phase transition for these liposomes. Binding of the cytidylyltransferase to egg PC vesicles containing oleic acid, monoolein, oleoylacetylglycerol or diolein resulted in enzyme activation, as did binding to dipalmitoylPC-oleic acid vesicles. However, binding to egg PC-palmitic acid vesicles did not fully activate the transferase. Various mechanisms for cytidylyltransferase interaction with membranes are discussed. PMID- 3032270 TI - Effects of vasopressin on the synthesis of phosphatidylethanolamines and phosphatidylcholines by isolated rat hepatocytes. AB - The effect of vasopressin on the biosynthesis of phosphatidylcholines and phosphatidylethanolamines was investigated in freshly isolated rat hepatocytes in suspension. Treatment of hepatocytes with vasopressin inhibits the incorporation of [Me-14C]choline into phosphatidylcholines in a dose-dependent manner. The hormone does not affect the uptake, phosphorylation or oxidation of choline. Pulse-chase studies indicate that CTP:cholinephosphate cytidylyltransferase might be subject to hormonal regulation by vasopressin. In contrast with the inhibitory effect of vasopressin on the synthesis of phosphatidylcholines, this hormone stimulates the incorporation of [1,2-14C]ethanolamine into phosphatidylethanolamines in a dose-dependent manner. Pulse and pulse-chase studies with labelled ethanolamine show that the conversion of ethanolaminephosphate to CDPethanolamine as well as the formation of phosphatidylethanolamines from CDPethanolamine and diacylglycerol are enhanced. Determination of the effect of vasopressin on the activity of the enzymes of the synthesis de novo of phosphatidylethanolamines demonstrates an increase of the activity of ethanolaminephosphotransferase, probably as a result of the increased amount of diacylglycerol in vasopressin-treated cells. PMID- 3032271 TI - Di(2-ethylhexyl)phthalate enhances hepatic phospholipid synthesis in rats. AB - The effects of di(2-ethylhexyl)phthalate, a typical peroxisomal proliferator, on the activities of key enzymes in the glycerophospholipid synthetic pathway and the incorporation of lipid precursors into liver lipids in vitro were studied periodically in rats. When di(2-ethylhexyl)phthalate was fed at the 1% level to rats, glycerol-3-phosphate acyltransferase activity increased 2-3-fold in liver homogenates and microsomes in 2-4 days. The specific activity of microsomal CTP:phosphocholine cytidylyltransferase increased by 1.5-fold, whereas the cytosolic activity was depressed. The microsomal CDPcholine:diacylglycerol cholinephosphotransferase specific activity decreased, whereas the activity in the homogenates increased, suggesting the proliferation of the hepatic endoplasmic reticulum in di(2-ethylhexyl)phthalate-treated rats. The incorporation of [1(3)-3H]glycerol or [1-14C]acetate into liver phospholipids in vitro increased in 2 days and stayed at a high level up to 12 days. The present study confirmed that di(2-ethylhexyl)phthalate induced an enhancement of phospholipid synthesis in the liver. The increase in hepatic phospholipid synthesis by this drug is presumably linked to the proliferation of peroxisomes and other intracellular membranes. PMID- 3032272 TI - Activation of hydrazine derivatives to free radicals in the perfused rat liver: a spin-trapping study. AB - Spin-trapping techniques and electron spin resonance spectroscopy have been used to study bioactivations of hydrazine and its derivatives by isolated perfused rat livers. Using phenylbutylnitrone (PBN) as the stable spin trap, it was found that the liver perfusion of hydrazine, acetylhydrazine and isoniazid resulted in the formation of the same carbon-centered radical which was shown to be the acetyl radical. The identity of the acetyl radical was confirmed after organic extraction of the liver perfusates, by comparing its coupling constants with those of the in vitro metal ion- or horseradish peroxidase-catalyzed oxidation products of the hydrazines in the same solvents. The liver perfusion of iproniazid formed the isopropyl radical which was previously observed to result from peroxidase-catalyzed oxidation. PMID- 3032273 TI - Purification and some properties of liver adenylylsulfate kinase. AB - Adenylylsulfate kinase (ATP:adenylylsulfate 3'-phosphotransferase, EC 2.7.1.25) has been purified over 1300-fold from rat liver in 10% yield. The enzyme has a molecular weight of 58,000 and is composed of four subunits of equal molecular weight. ATP is an allosteric activator of adenylylsulfate kinase, with a Hill coefficient of 2.2 and a K0.5 of 2.5 mM. Adenosine phosphosulfate is a potent inhibitor of adenylylsulfate kinase, but the adenosine phosphosulfate concentration for maximal reaction is dependent on the ATP concentration. At the physiological levels of ATP the inhibition by adenosine phosphosulfate is not likely to play a role, while the allosteric regulation of adenylylsulfate kinase by ATP may be operative. PMID- 3032275 TI - The oxidation of NADH by vanadate plus sugars. AB - Sugars and sugar phosphates enable vanadate to catalyze the oxidation of NADH. Superoxide dismutase inhibits this oxidation. Incubation of sugars with vanadate, prior to addition of NADH, accelerates this oxidation of subsequently added NADH and eliminates the lag phase otherwise noted. Incubation of sugars with vanadate also results in the reduction of vanadate to vanadyl, with appearance of a blue green color probably associated with a vanadyl-vanadate complex. It appears that sugars reduce vanadate to vanadyl which, in turn, reduces O2 to O2- and that vanadate plus O2- then catalyzes the oxidation of NAD(P)H by a free radical chain reaction. Such oxidation of NAD(P)H may account for several of the biological effects of vanadate. PMID- 3032274 TI - Nucleoside triphosphate pyrophosphatase of rabbit matrix vesicles, a mechanism for the generation of inorganic pyrophosphate in epiphyseal cartilage. AB - Inorganic pyrophosphate (PPi) may be important in the regulation of mineralisation but its origin in epiphyseal cartilage is ill-defined. Nucleoside triphosphate pyrophosphatase is one potential source, as this enzyme catalyses the formation of PPi from nucleoside triphosphates. This enzyme has been identified in matrix vesicles derived from rabbit epiphyseal cartilage and a method developed to measure the activity using ATP as substrate in intact matrix vesicles under relatively physiological conditions. The enzyme had a high affinity for ATP (Km less than 10 microM) and was also active towards GTP, CTP and UTP. Disruption of the matrix vesicle membrane by sonication failed to alter the activity. Treatment of sonicated matrix vesicles with Triton X-100 increased the activity which may indicate a direct effect of the detergent on the enzyme. Activity towards ATP was inhibited substantially by ADP and AMP and by another potential substrate beta,gamma-methyleneadenosine 5'-triphosphate. Dichloromethylene bisphosphonate, an analogue of the product PPi, inhibited the activity to a lesser extent. Two other potential substrates, NADP+ and thymidine 5'-monophosphate p-nitrophenyl ester were only weakly inhibitory as was 1 hydroxyethylidene 1,1-bisphosphonate. These results imply that nucleoside triphosphates are the substrates in vivo and the inhibitory effects of ADP and AMP suggest mechanisms whereby this activity could be regulated. PMID- 3032276 TI - A dual role for calcium in regulation of superoxide generation by stimulated rat alveolar macrophages. AB - Superoxide production in alveolar macrophages is stimulated by agonists which act through Ca2+-mediated (concanavalin A) and/or protein kinase C (phorbol ester or diacylglycerol analogues) -mediated events. Simultaneous addition of saturating concentrations of concanavalin A and a protein kinase C activator (either phorbol 12-myristate-13-acetate or 1-oleoyl-2-acetyl-sn-glycerol) caused a supra-additive enhancement of the initial rate of O2-. production. This synergism closely correlated with the known time-course of Ca2+ mobilization induced by concanavalin A; however, it occurred under conditions in which protein kinase C activation is reportedly not Ca2+ dependent. Phorbol ester-induced O2-. production was partially inhibited by the Ca2+ ionophore, A23187. Although phorbol ester-stimulated O2-. production initially was enhanced by concanavalin A, the duration of this O2-. production was reduced in comparison to that induced by phorbol ester alone. These results suggest a dual role for intracellular Ca2+ in both stimulatory and inhibitory regulation of O2-. production. PMID- 3032277 TI - Modulation of alkaline phosphodiesterase I in cultured rat hepatocytes. AB - Alkaline phosphodiesterase I activity was measured in adult and foetal rat hepatocytes maintained in primary culture under various conditions. This enzyme was found to be expressed in both cell populations and could be resolved into two bands having apparent molecular weights of 130,000 and 250,000, respectively. Alkaline phosphodiesterase I activity was already at high levels in 15 day foetal liver and, as early as the 19th day of gestation, it reached adult levels. Alkaline phosphodiesterase I levels were well maintained during culture. In the absence of serum, its level continued to increase with time in foetal cells. It dramatically increased by days 4 and 5, in adult cells maintained on fibronectin and plastic, respectively. Dexamethasone stimulated alkaline phosphodiesterase I activity after a lag phase of 8 h, with a maximum reached after 40 h. As this induction was prevented by addition of actinomycin D or cycloheximide, it could be concluded that it required RNA and protein synthesis. Only the major Mr 250,000 form responded to dexamethasone and was sensitive to serum. PMID- 3032279 TI - Identification of functional cAMP-dependent protein kinase in a 'neurite minus' mouse neuroblastoma cell line. AB - We have characterized and quantitated the level of cAMP-dependent protein kinase in the NS-20, N1E-115, N-18 and N1A-103 mouse neuroblastoma clonal cell lines, and we have correlated the occurrence of functional cAMP-dependent protein kinase with the dibutyryl cAMP-induced differentiated functions in these cells. Our results demonstrate the presence of functional cAMP-dependent protein kinase in extracts of all four cell lines examined, including the 'neurite minus' N1A-103 cell line. Dibutyryl cAMP induced neurite outgrowth and acetylcholinesterase activity in the NS-20, N1E-115 and N-18 neuroblastoma cell lines, but not in the N1A-103 cell line. However, dibutyryl cAMP caused a 2-3-fold increase in the R1 regulatory subunit protein and cAMP-phosphodiesterase activity in the 'neurite minus' N1A-103 cells in a manner similar to that of the other three 'neurite positive' cell lines. These results suggest that the biochemical lesion(s) subserving the neurite-minus phenotype of the N1A-103 cells may be distal to the activation of cAMP-dependent protein kinase and is in a biochemical pathway distinct from the induction of R1 regulatory subunit protein and cAMP phosphodiesterase activity. PMID- 3032278 TI - Actions of cholera toxin on dispersed Chief cells from guinea pig stomach. AB - Although much is known about the actions of cholera toxin on intestinal and extra gastrointestinal tissues, almost nothing is known about the interaction of this toxin with cells in the stomach. In the present study, we prepared 125I-labeled cholera toxin (1900 Ci/mmol) and examined the binding of this radioligand to dispersed Chief cells from guinea pig stomach. Moreover, we examined the actions of cholera toxin on cellular cAMP and pepsinogen secretion from Chief cells. Binding of 125I-labeled cholera toxin could be detected within 5 min, was maximal by 60 min, and was increased by increasing the radioligand or cell concentrations. Inhibition of binding by unlabeled toxin indicated a dissociation constant of 3 nM and 8.7 X 10(5) cholera toxin receptors per Chief cell. In contrast to the rapidity of binding, a cholera toxin-induced increase in cAMP and pepsinogen secretion was not detected until 30-45 min of incubation. A 3 to 6 fold increase in cAMP and pepsinogen secretion was observed with maximal concentrations of cholera toxin. Binding of 125I-labeled cholera toxin and the toxin's actions on cAMP and pepsinogen secretion were inhibited by the B subunit of the toxin. Binding was not altered by other agents that have been shown to stimulate pepsinogen secretion (carbachol, CCK-8, secretin, vasoactive intestinal peptide, prostaglandin E1, or forskolin). These data indicate that Chief cells from guinea pig stomach possess a specific class of cholera toxin receptors. Binding of cholera toxin to these receptors causes an increase in cellular cAMP that stimulates pepsinogen secretion. PMID- 3032280 TI - Association of low-density lipoprotein with particulate connective tissue matrix components enhances cholesterol accumulation in cultured subendothelial cells of human aorta. AB - Low-density lipoproteins (LDL) were incubated with elastin particles, collagenase resistant debris isolated from human aorta, and latex beads of 1.13 microns in diameter. As a result of incubation, insoluble LDL-associates were formed. These associates, as well as LDL-heparin-fibronectin-gelatin complexes described by other workers, were added to a 7-day primary culture of enzyme-isolated cells of human aortic subendothelial intima. The culture contained a mixed cell population made up mostly of typical and modified smooth muscle cells. 24 h later, total cholesterol, phospholipid, triacylglycerol, free cholesterol and cholesteryl ester levels were measured. Addition of insoluble LDL-complexes as well as LDL associates to culture brought about a substantial accumulation of intracellular lipids; primarily, cholesteryl esters. The total cholesterol level in cultured cells was raised 3- to 8-fold. Addition of free LDL or LDL-free particles had no effect on the content of intracellular lipids. The results obtained allow the assumption that the occurrence of the LDL-mediated accumulation of intracellular lipids is due mainly to the LDL penetration inside the cell via 'nonspecific' phagocytosis and not through a regulated receptor-dependent pathway. PMID- 3032282 TI - Vasopressin does not hydrolyze polyphosphoinositides in rabbit papillary collecting tubule cells. AB - Changes in phosphatidylinositol metabolism are suggested to be involved in the mechanism of action of many membrane active hormones. We studied the effect of vasopressin on polyphosphoinositide metabolism in rabbit papillary collecting tubule cells to assess if the hydrolysis of these phospholipids is involved in transmembrane signaling. Rabbit papillary collecting tubule cells grown in monolayers for 5 days were labeled to constant specific activity with [3H]inositol. The temporal changes in [3H]inositol-labeled phospholipids were assessed in response to vasopressin. Similarly, water-soluble inositides were monitored after separation by ion exchange chromatography. Intracellular Ca2+ was monitored by use of the fluorescent indicator dye, quin2. Vasopressin (10(-7) M) did not increase the hydrolysis of phosphoinositides over a 5 min period when compared with controls. Similarly, there was no increase in water-soluble phosphoinositols during the same interval. Pretreating the cells with LiCl (10 mM) did not produce any increase in inositol 1-phosphate when stimulated with vasopressin but did in response to bradykinin. Finally, vasopressin did not increase cytosolic Ca2+ and did not increase the release of prostaglandin E2 into the media under our experimental conditions. We conclude that vasopressin does not stimulate prostaglandin E2 in rabbit papillary collecting tubule cells, does not initiate hydrolysis of polyphosphoinositides and does not increase cytosolic Ca2+. Thus these cells lack V1 receptor coupling mechanisms. PMID- 3032281 TI - Stimulatory effect of glucose 6-phosphate on the non-mitochondrial Ca2+ uptake in permeabilized hepatocytes and Ca2+ release by inositol trisphosphate. AB - The relationships between Ca2+ transport and glucose-6-phosphatase activity, previously studied in isolated liver microsomes, were investigated in permeabilized hepatocytes in the presence of mitochondrial inhibitors. It was found that the addition of glucose 6-phosphate to the cells markedly stimulates the MgATP-dependent Ca2+ uptake. A progressive increase in the stimulation of Ca2+ uptake was seen with increasing amounts of glucose 6-phosphate up to 5 mM concentrations. Vanadate, when added in adequate concentrations (20-40 microM) to the hepatocytes inhibits both the glucose-6-phosphatase activity and the stimulation of Ca2+ uptake by glucose 6-phosphate, while not affecting the MgATP dependent Ca2+ uptake. The addition of inositol 1,4,5-trisphosphate to permeabilized hepatocytes in which Ca2+ had been accumulated in the presence of MgATP and glucose 6-phosphate, results in a rapid release of Ca2+. PMID- 3032283 TI - Dephosphorylation of cardiac myofibril C-protein by protein phosphatase 1 and protein phosphatase 2A. AB - C-protein purified from chicken cardiac myofibrils was phosphorylated with the catalytic subunit of cAMP-dependent protein kinase to nearly 3 mol [32P]phosphate/mol C protein. Digestion of 32P-labeled C-protein with trypsin revealed that the radioactivity was nearly equally distributed in three tryptic peptides which were separated by reversed-phase HPLC. Fragmentation of 32P labeled C-protein with CNBr showed that the isotope was incorporated at different ratios in three CNBr fragments which were separated on polyacrylamide gels in the presence of sodium dodecyl sulfate. Phosphorylation was present in both serine and threonine residues. Incubation of 32P-labeled C-protein with the catalytic subunit of protein phosphatase 1 or 2A rapidly removed 30-40% of the [32P]phosphate. The major site(s) dephosphorylated by either one of the phosphatases was a phosphothreonine residue(s) apparently located on the same tryptic peptide and on the same CNBr fragment. CNBr fragmentation also revealed a minor phosphorylation site which was dephosphorylated by either of the phosphatases. Increasing the incubation period or the phosphatase concentration did not result in any further dephosphorylation of C-protein by phosphatase 1, but phosphatase 2A at high concentrations could completely dephosphorylate C protein. These results demonstrate that C-protein phosphorylated with cAMP dependent protein kinase can be dephosphorylated by protein phosphatases 1 and 2A. It is suggested that the enzyme responsible for dephosphorylation of C protein in vivo is phosphatase 2A. PMID- 3032284 TI - Subcellular distribution of alpha 1-adrenergic receptors in rat liver. AB - The distribution of alpha 1-adrenergic receptors in rat liver subcellular fractions was studied using the alpha 1-adrenergic receptor ligand [3H]prazosin. The highest number of [3H]prazosin binding sites was found in a plasma membrane fraction followed by 2 Golgi and a residual microsomal fraction, the numbers of binding sites were 1145, 845, 629 and 223 fmol/mg protein, respectively. When the binding in these fractions was compared with the activity of plasma membrane 'marker' enzymes in the same fractions a relative enrichment of [3H]prazosin binding sites was found in the residual microsomes and one of the Golgi fractions. Photoaffinity labelling with 125I-arylazidoprazosin in combination with SDS-polyacrylamide gel electrophoresis revealed the specific binding to 40 and 23 kDa entities in a Golgi fraction, while in plasma membranes the binders had an apparent molecular mass of 36 and 23 kDa. When [3H]prazosin was injected in vivo into rat portal blood followed by subcellular fractionation of liver, a pattern of an initial rapid decline and thereafter a slow decline of radioactivity was noted in all fractions. Additionally, in the two Golgi fractions a transient accumulation of radioactivity occurred between 5 and 10 min after the injection. The ED50 values for displacement of [3H]prazosin with adrenaline was lowest in the plasma membrane fraction, followed by the residual microsomes and Golgi fractions, the values were 10(-6), 10(-5) and 10(-4) mol/l, respectively. On the basis of lack of correlation between distribution of alpha 1 adrenergic antagonist binding and adenylate cyclase activity, differences in the molecular mass of alpha 1-adrenergic antagonist binders, differences in the kinetics of in vivo binding and accumulation of [3H]prazosin and also differences in agonist affinity between plasma membrane and Golgi fractions, it is concluded that alpha 1-adrenergic receptors are localized to low-density intracellular membranes involved in receptor biosynthesis and endocytosis. PMID- 3032285 TI - Malondialdehyde production from spermine by homogenates of normal and transformed cells. AB - The oxidation of spermine in vitro by a mixture of polyamine oxidase and diamine oxidase from pig kidney gives rise to malondialdehyde via 3-aminopropanol as the intermediate. Conversely, with spermidine, under similar experimental conditions, no evidence could be obtained for malondialdehyde formation within the limits of sensitivity of the assay (2.0 nmol). The activities of both these enzymes show about a 2-fold increase in normal rat kidney cells (LA31 NRK) transformed by the temperature sensitive mutant of Rous sarcoma virus (LA31) and incubated at the non permissive temperature (39 degrees C) compared to the activities in LA31 NRK at the permissive temperature (33 degrees C). These same enzymatic activities show no temperature dependent changes in normal rat kidney cells (NRK) or in these same cells infected by the wild type virus (NRK B77). In extracts derived from Friend erythroleukemic cells induced to differentiate by dimethyl sulfoxide or hexamethylene bis acetamide, spermine oxidation takes place more efficiently than in non induced cells. A rise in diamine oxidase activity is seen in LA31 NRK (39 degrees C) 12 h after the temperature shift, whereas morphological manifestations of normalcy are seen only at 48 h. The Km of diamine oxidase is 10(-6) M for putrescine and 10(-3) M for 3-aminopropanol. A possible mechanism involving the well documented acetylation of putrescine [23,26] is proposed for diverting intracellular putrescine away from cytosolic diamine oxidase and towards intramitochondrial monoamine oxidase. PMID- 3032286 TI - Effects of butyric and acetic acids on acetone-butanol formation by Clostridium acetobutylicum. AB - The actions of butyric and acetic acids on acetone-butanol fermentation are investigated. Production of butyric and acetic acids are controlled by the extracellular concentrations of both acids: acetic acid added to the medium inhibits its own formation but has no effect on butyric acid formation, and added butyric acid inhibits its own formation but not that of acetic acid. The ratio of end metabolites depends upon acetic and butyric acid quantities excreted during the fermentation. In contrast to acetic acid, which specifically increases acetone formation, butyric acid increases both acetone and butanol formations. Acetate and butyrate kinase activities were also examined. Both increase at the start of fermentation and decrease when solvents appear in the medium. Coenzyme A transferase activity is weak in the acidogenic phase and markedly increases in the solvent phase. Acetic and butyric acids appear to be co-substrates. On the basis of these results, a mechanism of acetic and butyric acid pathways, coupled to solvent formation by C. acetobutylicum glucose fermentation is proposed. PMID- 3032287 TI - [Relation between Mg2+ activation and AMP inhibition of fructose-1,6 diphosphatase from rabbit skeletal muscles]. AB - It was shown that AMP, an allosteric inhibitor of fructose-1.6-bisphosphatase, decreases the apparent affinity of the enzyme for the activating cation, Mg2+, which is accompanied by a decrease of the kinetic cooperativity between the Mg2+ binding sites. In its turn, the Mg2+ increase diminishes the enzyme sensitivity to the inhibiting effect of AMP and decreases the cooperativity of the inhibitor binding. The heterotropic interactions between the allosteric inhibitor and activator binding centers are consistent with the predictions of the Monod-Wyman Changeux model which involves two conformational states of the enzyme (of which one is catalytically inactive) differing in their affinity for the ligands. An increase in pH from 7.4 to 9.0 increases the enzyme affinity for Mg2+ and causes an equilibrium shift towards the catalytically active state of the enzyme. PMID- 3032288 TI - [The use of 1H-NMR-spectroscopy for the study of peptide degradation by angiotensin-converting enzyme]. AB - A new method for continuous registration of enzymatic hydrolysis of peptides involving 1H-NMR spectroscopy was developed. The advantages of the method were demonstrated, using dalargin (Tyr-D-Ala-Gly-Phe-Leu-Arg) hydrolysis catalyzed by human kidney angiotensin-converting enzyme as an example. It was shown that the maximal activity of the enzyme towards dalargin is observed at pH 7.8; Km is 0.35 mM. The enzyme is inhibited by the substrate (Kd = 0.55 mM). Cl- do not influence the catalytic activity of the enzyme with respect to dalargin. The stereospecificity of the angiotensin-converting enzyme towards dalargin diasteriomers was studied. PMID- 3032289 TI - Hypothalamic peptide modulation of EEG sleep in depression: a further application of the S-process hypothesis. PMID- 3032290 TI - Distribution and levels of cellular retinol- and cellular retinoic acid-binding protein in various types of rat testis cells. AB - The distribution and levels of cellular retinol-binding protein (CRBP) and cellular retinoic acid-binding protein (CRABP) were measured in rat testicular peritubular and Sertoli cells and in isolated rat pachytene spermatocytes and spermatids. Two Sertoli cell preparations, one containing some germ cells and another that had been osmotically shocked to destroy germ cells, were examined. CRBP and CRABP levels were measured by specific and sensitive radioimmunoassays. Testicular peritubular cell cytosol preparations were found to contain high levels of CRBP (1.48 +/- 0.87 microgram CRBP/mg protein) but CRABP could not be detected. The mean CRBP level in Sertoli cell preparations that contained some germ cells was 0.93 +/- 0.24 microgram CRBP/mg protein; this value was similar to the level of 1.11 +/- 0.20 microgram CRBP/mg protein measured for Sertoli cells free of germ cells. The level of CRABP found in Sertoli cell preparations containing germ cells (0.81 +/- 0.32 microgram CRABP/mg protein) was approximately five times greater than was observed in Sertoli cells free of germ cells (0.16 +/- 0.03 microgram CRABP/mg protein). CRBP and CRABP levels in cultured Sertoli cells were not affected by time in culture for up to five days of culture. Pachytene spermatocytes and spermatids were very enriched in CRABP (0.72 +/- 0.26 microgram CRABP/mg protein for spermatocytes and 0.65 +/- 0.21 microgram CRABP/ml protein for spermatids). A search for a high molecular weight retinol-binding protein did not demonstrate the existence of such a protein in Sertoli cell-conditioned medium. In summary, these studies provide quantitative information about the distribution of the cellular retinoid-binding proteins in the cell types that compose the rat testis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3032291 TI - The molecular basis of gonadotropin-releasing hormone (GnRH) action in the pituitary gonadotrope. PMID- 3032292 TI - Early effects of luteinizing hormone on mitochondrial phosphoinositides in ovarian follicles. AB - It is not clear if luteinizing hormone (LH) stimulates breakdown as well as synthesis of phosphoinositides in ovarian tissue. Possibly, LH stimulation results in hydrolysis of ovarian phosphoinositides in discrete subcellular compartments while increasing their synthesis at other sites. To investigate this hypothesis, we determined the effects of LH on phosphoinositide metabolism in whole homogenates and mitochondria of ovarian follicles. Medium (3-7 mm) follicles from porcine ovaries were preincubated for 2 h in phosphate (PO4)-free medium with 32PO4, and incubated without or with LH (1 microgram/ml). Phosphatidylinositol (PI) and related compounds, phosphatidic acid (PA), phosphatidylinositol phosphate (PIP) and phosphatidylinositol bisphosphate (PIP2), accounted for 40% of the radiolabeled phospholipids in whole homogenates and over 60% in mitochondria from preincubated follicles. After 5 min, LH caused a significant decrease in radiolabeling of PIP2 and PIP in mitochondria, but not in whole homogenates. Luteinizing hormone increased radiolabeling of PIP2, PIP, PI and PA within 10 min in whole homogenates, and within 20 to 30 min in mitochondria. This delayed increase in radiolabeling of mitochondrial phosphoinositides after LH treatment was accompanied by decreases in PIP2, PIP and PI radiolabeling in whole homogenates. Follicles also were preincubated for 4 h with [3H]inositol, then for 15 min with 10 mM LiCl (an inhibitor of inositol phosphate hydrolysis). Inositol phosphate accumulation in 30 min was 2.7 times higher in homogenates of LH-treated follicles then in untreated follicles. Also, LH significantly decreased inositol bisphosphate, but did not change inositol trisphosphate accumulation. Accumulation of inositol phosphates in mitochondria was not measurable.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3032293 TI - Electron density profile of two-dimensionally crystalline membranous cytochrome c oxidase at low resolution. AB - Unilamellar vesicles of membranous cytochrome c oxidase have been isolated whose distribution of protein in the membrane plane was predominantly crystalline. The vesicles were collapsed via controlled partial dehydration, resulting, at first, in the formation of unoriented, mostly unstacked, membrane pairs. Further controlled partial dehydration resulted in the formation of oriented multilayers of stacks of membrane pairs, retaining the in-plane crystallinity. The above were monitored by electron microscopy and x-ray diffraction. Analysis of the x-ray diffraction from unoriented, unstacked membrane pairs by two independent methods provided the membrane electron density profile to 30 A resolution. PMID- 3032294 TI - Fluorescence quenching dynamics of tryptophan in proteins. Effect of internal rotation under potential barrier. AB - In many proteins fluorescence from single tryptophan exhibits a nonexponential decay function. To elucidate the origin of this nonexponential decay, we have examined the fluorescence decay function and time-resolved fluorescence anisotropy of a fluorophore covalently bound to a macromolecule by solving a rotational analogue of the Smoluchowski equation. An angular-dependent quenching constant and potential energy for the fluorophore undergoing internal rotation were introduced into the equation of motion for fluorophore. Results of numerical calculations using the equations thus obtained predict that both the fluorescence decay function and time-resolved anisotropy are dependent on rotational diffusion coefficients of fluorophore and potential energy for the internal rotation. The method was applied to the observed fluorescence decay curve of the single tryptophan in apocytochrome c from horse heart. The calculated decay curves fit the observed ones well. PMID- 3032295 TI - Correlation between computed conformational properties of cytochrome c peptides and their antigenicity in a T-lymphocyte proliferation assay. PMID- 3032296 TI - A proton NMR spin-lattice relaxation study of the imino proton exchange kinetics in calf-thymus DNA. PMID- 3032297 TI - [The role of the adenylate cyclase system in the inhibition of gastric secretion by products of limited hydrolysis of kappa-casein]. AB - The influence of the peptide inhibiting gastric acid secretion and derived from kappa-casein on the levels of cAMP, cGMP and adenylate cyclase activity of rat gastric mucosa was studied. Intravenous peptide preparation administration failed to alter cyclic nucleotide levels. The revealed decrease of adenylate cyclase activity was due to the presence of calcium in the investigated peptide preparation. It was assumed that cyclase system of the rat gastric mucosa cells were not involved in the inhibition of gastric acid secretion by the peptide preparations under study. PMID- 3032298 TI - [Blood levels of corticosterone, aldosterone, angiotensin and ACTH in rats after deafferentation of the tongue]. AB - The influence of tongue deafferentation on the plasma level of sodium homeostasis controlling hormones has been studied. Using radioimmunoassay, high corticosterone and aldosterone plasma levels were discovered in rats with tongue deafferentation, as compared to sham-operated controls. ACTH and angiotensin I plasma concentrations in deafferentated rats were the same as in sham-operated rats. The role of emotional factors of taste perception in sodium homeostasis control is being discussed. PMID- 3032299 TI - [Effect of cholera enterotoxin on carbohydrate metabolism of the liver and small intestine mucosa in the rabbit]. AB - Changes in carbohydrate metabolism were studied in the isolated intestinal loops of rabbits during secretory diarrhea, induced by cholera enterotoxin. Glucose synthesis level in the small intestinal mucosa and liver was measured by isotope technique, using L-alanine as a precursor. Intestinal gluconeogenesis, calculated per mg of protein, appeared to be twice higher than in the liver of fasting rabbits. Cholera enterotoxin administration enhanced gluconeogenesis in the liver by 60%, as compared to the control. The rate of glucose synthesis and glucose-6 phosphatase activity in the intestinal mucosa remained unchanged, whereas glucose 6-phosphatase in the liver was slightly inhibited. It is suggested that gluconeogenesis in the liver supplies glucose as a convenient energy source for the secretory process induced by cholera enterotoxin in the rabbit small intestine. PMID- 3032300 TI - [Phosphatidylcholine-induced repair of damaged hepatocyte membranes in heliotrine poisoning]. AB - Hepatocyte membranes destruction in experimental toxic hepatitis caused by heliotrine administration was accompanied by a 10-fold increase in blood serum activity of aldolase fructose-I-monophosphate, a decrease in cytochrome P-450 content, an increase in the rate of cytochrome P-450 inactivation, as well as a decrease in microsomal glucose-6-phosphatase activity. Administration of phosphatidylcholine liposomes decreased the activity of aldolase twofold, which indirectly shows partial reconstitution of liver cell membranes. Phosphatidylcholine protective action is also manifested in an increase in the activity of glucose-6-phosphatase, a microsomal marker enzyme, up to its control level and in a 20% reduced rate of cytochrome P-450 inactivation. It has been shown that destroyed liver cell membranes may be repaired by the introduction of phosphatidylcholine in the form of multilayer liposomes. PMID- 3032301 TI - [The role of various opiate receptors in the regulation of the nociceptive reaction of the arterial pressure]. AB - The experiments on alert rats have shown that dissociation in opioid regulation of behavioural and hemodynamic pain manifestations is determined at a spinal opiate receptor level. Opiates and opioids suppress blood pressure nociceptive reactions to mu-opiate receptors, while sigma-opiate receptors are involved into the generation of autonomic activating effect in opiate analgesia. PMID- 3032302 TI - [The effect of opiate receptor ligands on emotional cardiovascular reactions in lower primates]. AB - Intravenous naloxone injection (0.1 mg/kg) facilitated blood pressure increase in response to conditioned sound stimulus followed by electrocutaneous shock in conscious chair-restrained baboons (Papio hamadryas). Naloxone at a dose of 1.0 mg/kg had an opposite effect and led to the decrease in blood pressure and heart rate in conditioned fear reflex. Naloxone microinjections (50 microM) into the periventricular hypothalamus led to a significant diminution of blood pressure and heart rate increment in response to electrocutaneous shock; naloxone microinjections into tractus solitarius nuclei suppressed blood pressure and heart rate reactions both to conditioned (sound) and unconditioned (electrocutaneous shock) stimuli. Microinjections of equimolar morphine quantities in these brain regions facilitated such reactions. It is concluded that endogenous opioid system participates in the formation of cardiovascular reactions to emotional stimuli in monkeys, with multiple opioid receptors of periventricular hypothalamus and tractus solitarius nuclei involved in the generation of such reactions. PMID- 3032303 TI - [The site (central or peripheral) of the anti-ulcer action of dalargin, a synthetic analog of endogenous opioids in an experimental model of cysteamine induced duodenal ulcer in rats]. AB - Dalargin injected subcutaneously at a dose of 10 micrograms/kg decreased 4-5 fold ulcer manifestations in rats with cysteamine-induced duodenal ulcers. Intracerebroventricular dose of 2 micrograms diminished the manifestations to a lesser extent Dalargin only at a dose exceeding 500 micrograms intraperitoneally decreased significantly the in vivo binding of 3H-D-Ala2, D-Leu5-enkephalin with brain opiate receptors. We believe that Dalargin injected peripherally in small doses does not penetrate the blood-brain barrier and that its antiulcer activity is due to the interaction with peripheral opioid receptors. It seems possible that the disturbances in the central/peripheral ratio of the opioid activity plays an important role in the pathogenesis of duodenal peptic ulcer. PMID- 3032304 TI - [Ultrastructural and functional changes in the myocardium and coronary vessels after massive blood loss]. AB - The ultrastructure of cardiomyocytes and circulatory bed has been compared to transmembrane cAMP-dependent Ca2+ transport in experiments on the hearts of 14 dogs immediately after massive blood loss. The results an hour after non compensatory hemorrhage have shown extra- and intracellular myocardial edema, central destruction of sarcomers, steep increase in the volume of agranular sarcomplasmic reticulum and T-system, different degree of damage of other organoids, and also disturbances in the ultrastructure of venous capillary and postcapillary section. The biochemical techniques used have shown a decrease in Ca2+ transporting ability of sarcolemma due to its AMP-dependent regulation of cardiomyocytes. Excessive Ca2+ storage in cytosole promoted the appearance of "constriction bands" in myofibrils. PMID- 3032305 TI - Ki-M8 monoclonal antibody reactive with an intracytoplasmic antigen of monocyte/macrophage lineage. AB - A monoclonal antibody (MoAb), Ki-M8, that reacts specifically with cells of the monocyte/macrophage system is described. On light and electron microscopic immunohistochemistry, Ki-M8 recognizes intracytoplasmatically localized antigens of mol wt 30,000 and 32,000, increasingly expressed during differentiation of monocytes into macrophages. Ki-M8 antigen is detectable on almost all known tissue macrophages and monocyte/macrophage-related cell lines after appropriate stimulation. In functional terms Ki-M8 significantly impairs the generation of oxygen radicals during an induced respiratory burst. Applied to acute nonlymphoblastic leukemias, a clear-cut differentiation of the monocytic phenotype and differentiation is possible on the basis of Ki-M8 immunoreactivity. Ki-M8 represents a reagent specific for the monocyte/macrophage system with regard to antigen distribution in normal and neoplastic cells as well as with regard to its influence on a typical monocyte/macrophage-related function. PMID- 3032306 TI - Fate of the DNA in plasmid-containing Escherichia coli minicells ingested by human neutrophils. AB - Escherichia coli minicells containing the plasmid pSC101 (approximately 10 kb) or pBR322 (approximately 4 kb) were opsonized and incubated with human neutrophils. The neutrophils responded to the minicells as they would to native E coli: they ingested the minicells, discharged their granule contents into the minicell containing phagosomes, and expressed a respiratory burst. After one hour of incubation, the fate of the ingested plasmid DNA was examined. No DNA degradation was detected by trichloroacetic acid precipitation or agarose gel electrophoresis. Moreover, when pBR322 recovered from ingested minicells was transformed into E coli, no mutations in either of the antibiotic resistance genes carried by the plasmid were detected out of many thousand transformants screened. These findings confirm the surprisingly limited effect of neutrophils on ingested DNA. PMID- 3032307 TI - Iron deficiency and neutrophil function: different rates of correction of the depressions in oxidative burst and myeloperoxidase activity after iron treatment. AB - The polymorphonuclear granulocyte (PMN) kills ingested bacteria by mechanisms that include myeloperoxidase (MPO) and a sudden increase in oxygen consumption (the oxidative burst), both of which are iron dependent. The magnitude of the oxidative burst and activity of MPO were determined in PMNs during the progression of iron deficiency (ID) and following its treatment in rats. As ID developed, the oxidative burst after zymosan activation was less depressed than the activity of MPO. There was no change in the oxidative burst after activation with phorbol myristate acetate (PMA) or in the generation of superoxide (O2-) by NADPH oxidase-containing particles from PMNs. Following iron treatment, impairment of the oxidative burst after zymosan activation was corrected after 1 day. In contrast, the deficit in MPO activity was not corrected until 7 days after initiation of iron treatment. The pattern of recovery in MPO activity after iron treatment corresponded to the prolonged period of maturation of the PMN primary granule since the formation of primary granules, which contain MPO, takes place only in the early, mitotic stages of maturation. The tendency of the PMN to maintain the oxidative burst allows the cell to preserve its capacity for bacterial killing during the progression of iron deficiency. PMID- 3032308 TI - Protein C inhibits endocytosis of thrombin-thrombomodulin complexes in A549 lung cancer cells and human umbilical vein endothelial cells. AB - We investigated the effect of protein C on the endocytosis of thrombin thrombomodulin complexes. We previously showed that exposure of umbilical vein endothelial cells to thrombin stimulated the internalization and degradation of thrombin. A similar internalization was stimulated by a monoclonal antithrombomodulin antibody. We have repeated these studies in the presence of protein C and found that endocytosis of 125I-thrombin-thrombomodulin complexes, but not 125I-antithrombomodulin-thrombomodulin complexes, is inhibited. Activated protein C did not inhibit endocytosis of thrombin-thrombomodulin complexes. Protein C inhibited both internalization and degradation of 125I-thrombin and diisopropylphosphoryl (DIP) 125I-thrombin in human lung cancer cells (A549). These effects were observed at protein C concentrations found in human plasma. Protein S had no effect on the inhibition of endocytosis of thrombin thrombomodulin complexes by protein C. We propose that protein C may regulate the rate of endocytosis of thrombin-thrombomodulin complexes in vivo and thereby control the capacity for endothelium to activate protein C. PMID- 3032309 TI - Human monoclonal antibody against Rh(D) antigen: partial characterization of the Rh(D) polypeptide from human erythrocytes. AB - A human monoclonal anti-Rh(D) antibody produced by an Epstein-Barr virus (EBV) transformed B-cell line (IgG1(lambda), clone H2D5D2) has been purified on protein A-Sepharose column and used for binding studies and immune precipitation of the blood group rhesus (Rh) antigens. Scatchard plot analyses show that the 125I labeled antibody (iodo-gen procedure), binds to 1.09 X 10(5), 0.43 X 10(5), and 0.32 X 10(5) antigen sites on each D--/D--, R2R2 and R1R1 RBC, respectively, with an association constant of approximately 0.6 X 10(8) mol/L-1. Immune precipitation studies indicate also that the Rh(D) antigen of the Rh(D)-positive RBCs is carried by a 29 kd polypeptide as deduced from sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). No material could be precipitated from Rh(D)-negative or Rhnull RBCs. These results indicate that the monoclonal and the polyclonal human anti-Rh(D) behave similarly. A sample (Blo., presumed genotype R2r or R0R2) showing an increased number of antigen sites (0.76 X 10(5)/cell) and a high binding constant (5.7 X 10(8) mol/L-1) was used, as well as D--/D-- RBCs, for further purification of the 29-kd component. Extraction by Triton X-100 (0.1% to 5%) of the immune complexes formed between the membrane bound Rh(D) antigens and the monoclonal antibody as well as a direct quantitative estimate of the 29-kd component, suggest that the Rh(D) polypeptide is loosely bound to the skeleton, since less than or equal to 80% can be solubilized from the membrane. In similar conditions, glycophorin A showed a slight association with the Triton-insoluble residue, whereas glycophorin B was easily and completely extracted. In contrast, both the minor RBC sialoglycoproteins, glycophorin C and glycoprotein gamma, remained predominantly bound to the membrane skeleton. The purified Rh(D) polypeptide obtained from Blo. and D--/D-- RBCs by immunoprecipitation and preparative gel electrophoresis was homogenous as judged by SDS-PAGE. Amino acid composition indicated that the Rh(D) protein contained sulfhydryl groups which are essential for biological activity. PMID- 3032310 TI - Synergistic inhibition of platelet activation by plasmin and prostaglandin I2. AB - Endothelial cell prostacyclin (PGI2) inhibits platelet activation by raising platelet cyclic AMP. Previously, platelet activation was also shown to be blocked by plasmin formed by endothelium-derived tissue plasminogen activator (TPA). We have now studied interactions between PGI2 and plasmin in the control of platelet function. PGI2 and plasmin cause synergistic inhibition of thrombin- and ADP induced aggregation of washed platelets. Inhibition by PGI2 is similarly potentiated by TPA added to platelet-rich plasma to generate plasmin. Thrombin stimulated rise in platelet cytosolic Ca2+, measured by fura2 fluorescence, and thromboxane A2 formation, measured by radioimmunoassay (RIA), are likewise synergistically inhibited by PGI2 and plasmin. Plasmin neither increases nor potentiates PGI2-stimulated increases in platelet cyclic AMP. Thus, PGI2 and plasmin cause synergistic inhibition of platelet activation by both cyclic AMP dependent and independent mechanisms. This interaction between two different endothelium-derived products may play an important role in localizing the hemostatic plug to a site of vascular injury by preventing further thrombin mediated accrual of platelets. PMID- 3032311 TI - Heterogeneity of mouse vascular endothelium. In vitro studies of lymphatic, large blood vessel and microvascular endothelial cells. AB - Microvascular endothelial cell cultures have been established from mouse lung, liver, brain, heart, placenta, kidney, urinary bladder, mammary gland, ovary and epididymal fat pad. In addition, large vessel endothelial cells have been obtained from the mouse aorta and thoracic duct. The heterogeneity of these cells has been shown by flow cytometric determination of angiotensin-converting enzyme, by differential presence of the acetyl low density lipoprotein receptor, by the variable expression of cell surface antigens, and by differential binding of various plant lectins. The endothelial cell lines we have developed provide the means to examine in the mouse, long a key species for biomedical research, a wide range of biological functions and properties of the vascular endothelium. PMID- 3032313 TI - Mass spectrometry of prostaglandins, leukotrienes and steroids as their allyldimethylsilyl ether derivatives. AB - Allyldimethylsilyl ethers of prostaglandin E2 and F2 alpha, leukotriene B4 and 5 beta-pregnane-3 alpha, 20 alpha-diol were prepared at room temperature in good yields with the novel reagent N,O-bis(allyldimethylsilyl)-trifluoroacetamide. The gas chromatographic properties of the derivatives of prostaglandins and the steroid were found to be excellent whereas those of leukotriene B4 were found to be less than satisfactory. The mass spectra of the allyldimethylsilyl ether derivatives of the compounds studied show intense ions in their upper mass region derived from the elimination of an allyl radical. In the mass spectrum of the derivative of leukotriene B4, the formation of a conjugated carbonium ion by alpha-cleavage is a major process. The detection limits for a quantitative assay were established by performing selected ion monitoring on the most intense ion in the upper mass region of respective mass spectrum. Detection limits in the low picogram range were obtained for the prostaglandins and the steroid but the allyldimethylsilyl ether derivative of leukotriene B4 exhibited a far higher detection limit. It is concluded for the latter compound that the detection limit depends primarily on the gas chromatographic properties. PMID- 3032312 TI - Stimulation of oxidative metabolism of granulocytes by recombinant granulocyte macrophage-colony-stimulating-factor and a conditioned medium of a urinary bladder carcinoma cell line. AB - Neutrophils (PMN) are the major host defence cells protecting the body against invasion by microorganisms. Products of oxidative metabolism mediate PMN microbicidal and tumoricidal activity, but the mechanisms by which these pathways become activated are not well understood. The colony stimulating factors (CSF) are known to stimulate proliferation and differentiation of committed bone marrow stem cells. These regulators may probably play an important role in non specific resistance to infections. We studied the oxidative metabolism of neutrophils after stimulation with recombinant GM-CSF (r.GM-CSF) and the concentrated conditioned medium of the UBC-5637 cell line (UBC-CM) showing CSF activity. It could be demonstrated that the r.GM-CSF, as well as the UBC-CM, induce an activation of the neutrophil respiratory burst without any cofactors such as f MLP, PMA, or zymosan. In addition, we observed an increase of the response to those stimulants in the presence of either r.GM-CSF or UBC-CM. These effects were not endotoxin-induced, since stimulation persisted after addition of Polymyxin B, which is known to inhibit the action of endotoxins. PMID- 3032314 TI - Replacement of aromatic fluorine by a methoxy group during reaction with methyl iodide in N,N-dimethylformamide solvent. AB - The DNA base uracil was derivatized with pentafluorobenzoyl chloride, followed by methylation with methyl iodide in the presence of N,N-dimethylformamide (DMF). In addition to a 3-pentafluorobenzoyl-1-methyl derivative of uracil, GC/MS analysis of the reaction mixture revealed the formation of an unusual product, whose molecular weight was 12 U higher than that of the prior derivative. This unexpected product has been identified as the 3-(para-methoxytetrafluorobenzoyl) 1-methyl derivative of uracil. Isotopic labeling and related experiments have revealed that the DMF solvent contributes the oxygen atom of the methoxy group that replaces the para fluorine atom. This work allowed a single derivative to be obtained for the methylation reaction by changing the solvent to acetonitrile. PMID- 3032315 TI - Viral alkaline nuclease in intranuclear dense bodies induced by herpes simplex infection. AB - The distribution of the herpes simplex virus alkaline nuclease (HSV DNase) in rabbit fibroblast cells infected with HSV type 1 or 2 (HSV-1 or HSV-2) has been examined with the aid of immunocytochemical techniques using gold particles as markers. HSV DNase was found to be accumulated within infected nuclei as early as 2.5 hr post-infection. Labeled antibody to HSV DNase subsequently increased in all nuclei after 7 hr and 17 hr. At all times post-infection the virus induced nuclear dense bodies were always the most intensely labeled structures. The association of HSV DNase with nucleoprotein containing structures was readily dissociated by hypotonic shock and detergent treatment, but its association with the dense bodies was not disrupted. Nucleolar 100 kDa protein was found to be simultaneously present in the dense bodies. This suggests that HSV DNase might interfere with ribosomal RNA synthesis and play a role in the degradation of the host-cell metabolism. PMID- 3032316 TI - Colchicine-induced tubular, vesicular and cisternal organelle aggregates in absorptive cells of the small intestine of the rat. II.--Endocytosis studies. AB - Administration of the antimicrotubular agent colchicine to adult rats (0.5 mg/100 g of body weight for 6 hr) induces formation of extended aggregates of tubular, vesicular, and cisternal organelles in the absorptive cells of the small intestine. The phosphatase reaction pattern (thiamine pyrophosphatase, acid phosphatase, acid trimetaphosphatase) suggests that the majority of them belongs to the lysosomal system (Ellinger and Pavelka, 1984). The present study extends these findings and examines the uptake and fate of intravenously injected horseradish peroxidase (HRP) at the basal and lateral cell surfaces and of intraluminally applied HRP at the apical cell surface. HRP, applied to control animals and animals pretreated with colchicine, was internalized at both apical and basolateral cell surfaces of the absorptive cells, and delivered into endosome-like vesicles, multivesiculated bodies (mvbs), dense bodies (dbs), and in several instances into Golgi cisternae. Following intraluminal application, evidence was obtained for the transport of HRP across the cell; in contrast, intravenously applied HRP was never detected at the apical cell surface. Colchicine pretreatment did not stop the uptake of HRP, which was rapidly sequestered to the clustered tubules, vesicles, and cisternae, as well as to the mvbs and dbs. After longer intervals, the portion of HRP-reactive tubules, vesicles, and cisternae within the clusters increased: 60 min after HRP administration all of them contained HRP-activity. These results indicate that the colchicine-induced clustered organelles are recipients of endocytic materials internalized at the apical as well as at the basolateral cell surface. PMID- 3032317 TI - Nuclear accumulation of HMG1 protein is correlated to DNA synthesis. AB - The subcellular localization of HMG1 protein was studied by immunoelectron microscopy during growth of CV1 cells in culture and in confluent CV1 cells subsequently lytically infected with SV40. HMG1 was always detected in the cytoplasm of both non-infected and infected cells. On the other hand, this protein displayed a nuclear localization only in those cells active in cellular and/or viral DNA replication, that is, in actively dividing non-infected cells and in confluent cells following SV40 infection. The combination of electron microscope immunocytochemistry and autoradiography revealed that during SV40 lytic infection, HMG1 accumulates at sites of active viral DNA replication. Since HMG1 is a single-stranded DNA binding protein and acts in vitro as a physiological nucleosome assembly factor, we suggest that its presence in the nucleus is related to its requirement in the DNA replication process. PMID- 3032318 TI - [Production of restriction endonucleases from various Salmonella strains of human origin]. PMID- 3032319 TI - Reconstruction of the cranial base following tumour resection. AB - Twenty-four patients have undergone resection of tumours involving the cranial base by a multidisciplinary team consisting of a neurosurgeon, ENT surgeon and plastic surgeon. The resultant defects of the cranial base have been reconstructed using local fascial flaps, transposition of local muscle flaps and microsurgical transfer of free muscle flaps. Indications for reconstruction have included obliteration of paranasal sinuses, coverage of tenuous dural repairs or dural grafts and separation of the nasopharynx from the dura of the frontal and temporal lobes and posterior fossa to prevent CSF leakage and meningitis. PMID- 3032320 TI - Modulation of GABAA receptor activity by alphaxalone. AB - The modulation of the gamma-aminobutyric acidA (GABAA) receptor by alphaxalone has been investigated by use of voltage-clamp recordings from enzymatically isolated bovine chromaffin cells maintained in cell culture. Alphaxalone (greater than 30 nM) reversibly and dose-dependently potentiated the amplitude of membrane currents elicited by locally applied GABA (100 microM). The potentiation was not associated with a change in the reversal potential of GABA-evoked currents and was not influenced by the benzodiazepine receptor antagonist, Ro15-1788 (300 nM). At relatively high concentrations (greater than 1 microM), alphaxalone directly elicited a membrane current. It is concluded that such currents result from GABAA receptor activation since they were reversibly suppressed by bicuculline (3 microM), dose-dependently enhanced by phenobarbitone (100-500 microM), and had a similar reversal potential (approximately 0 mV) to currents elicited by GABA. Additionally, on outside-out membrane patches, alphaxalone activated single channel currents with amplitudes and a reversal potential similar to those evoked by GABA. Alphaxalone (30 nM-1 microM) had no effect upon the amplitude of membrane currents elicited by locally applied acetylcholine (ACh) (100 microM). However, higher concentrations of alphaxalone (10-100 microM) reversibly suppressed ACh-evoked currents, the IC50 for blockade being 20 microM. The beta hydroxy isomer of alphaxalone, betaxalone (100 nM-1 microM), did not potentiate GABA-induced currents, nor did higher concentrations of the steroid (10-100 microM) directly evoke a membrane current. However, over the latter concentration range, betaxalone suppressed the amplitude of currents elicited either by GABA or ACh. The relevance of the present results to the anaesthetic action of alphaxalone is discussed together with the broader implications of steroidal modulation of the GABAA receptor. PMID- 3032321 TI - Changes in tissue blood flow and beta-receptor density of skeletal muscle in rats treated with the beta2-adrenoceptor agonist clenbuterol. AB - Rats injected with the beta2-adrenoceptor agonist clenbuterol (2 mg kg-1 per day) for 18 days gained significantly more weight than controls. Tissue blood flow assessed 24 h after the last injection from the distribution of radiolabelled microspheres was increased in white (5 fold) and brown (3 fold) adipose tissue of clenbuterol-treated rats but was unaffected in kidney, brain and diaphragm, and was reduced by about 80% in skeletal muscle. Acute injection of clenbuterol one hour before measuring blood flow, increased blood flow to brown fat (20 fold) in both treated and control groups. Blood flow to skeletal muscle increased more in the rats treated chronically with clenbuterol (6 fold increase) than in control rats (2 fold increase), but absolute flow rates were still significantly lower in the rats treated chronically with clenbuterol. Skeletal muscle beta-adrenoceptor density and subtype were assessed from ligand binding and displacement studies using [3H]-dihydroalprenolol. Rats treated with clenbuterol for 18 days showed a 50% reduction in beta-receptor density, but the ratio of beta 1/beta 2-receptors was unaffected (15% beta 1/85% beta 2). The results indicate that, although clenbuterol produces acute increases in muscle blood flow, chronic treatment results in lower flow rates immediately (1 h) and 24 h after the previous injection. The attenuated response following chronic treatment is associated with a marked reduction in skeletal muscle beta-adrenoceptor density. The data suggest that any anabolic effects of clenbuterol on muscle which may require, or may be mediated by increases in blood supply, cannot be sustained by chronic treatment. Conversely, blood flow to white and brown adipose tissue would appear to be potentiated by chronic treatment, possibly reflecting increases in lipolytic and/or thermogenic activity. PMID- 3032322 TI - A tumour-like bone lesion as a manifestation of vitamin D-resistant hypophosphataemic osteomalacia. PMID- 3032323 TI - Why women are not receiving anti-Rh prophylaxis. PMID- 3032324 TI - Angio-oedema and urticaria associated with angiotensin converting enzyme inhibitors. PMID- 3032325 TI - Dihydroxypropoxymethyl guanine in the treatment of AIDS related retinitis due to cytomegalovirus. PMID- 3032326 TI - Moderate sodium restriction with angiotensin converting enzyme inhibitor in essential hypertension. PMID- 3032327 TI - Eczema herpeticum: a potentially fatal disease. PMID- 3032328 TI - Hereditary generalized amyloidosis with polyneuropathy. Clinicopathological study of 65 Japanese patients. AB - A clinicopathological study was made on 65 patients from a small area of Nagano Prefecture, Japan, with hereditary generalized amyloidosis with polyneuropathy to clarify the clinical variety of the disease. Forty-five patients from Ogawa village showed similar clinical features. The age of onset ranged widely from 16 to 62 years. The main neurological manifestations were polyneuropathy starting in the legs and autonomic dysfunction. Lower cranial nerves were also affected in the advanced stages. Severe cardiac and renal involvement was uncommon. All these clinical features are consistent with type I familial amyloid polyneuropathy (FAP). The remaining 20 patients from five unrelated kinships showed unique clinical pictures. Two families from Ogawa village had type I FAP, but 4 out of the 5 affected patients showed marked nephropathy with heavy proteinuria from an early stage. Of the three other families, one, with 10 patients, was notable for the involvement of the central nervous system. Most of the patients showed cerebellar ataxia and pyramidal tract signs in addition to a sensorimotor and autonomic peripheral neuropathy. Another family had 2 siblings who had severe amyloid heart disease from the onset and developed polyneuropathy with autonomic features at an advanced stage. In the third family, onset occurred in the sixth decade in all 3 patients and the course was mild in 2, although the clinical features were those of typical type I FAP. Immunohistochemical study revealed that the amyloid fibril proteins in the patients with all four unusual clinical phenotypes were related to plasma prealbumin. The most common form of hereditary generalized amyloidosis in Japan is type I FAP, but the disease shows considerable variety in the age of onset and involves more systemic organs than previously recognized. The newly recognized clinical forms of hereditary generalized amyloidosis with severe amyloid heart disease or central nervous dysfunction indicate clinical heterogeneity of hereditary amyloidosis with polyneuropathy. PMID- 3032329 TI - Peripheral sensorimotor and autonomic neuropathy associated with systemic lupus erythematosus. Clinical, pathological and immunological features. AB - The clinical features and pathological findings in the sural nerves are described of 7 patients with peripheral neuropathy; in 4 cases the criteria for diagnosis of systemic lupus erythematosus (SLE) were satisfied and in 3 other cases there was serological evidence of an undifferentiated connective tissue disease, most probably SLE. The peripheral neuropathy was of a chronic sensorimotor type with predominantly sensory features and gradual onset. In 2 cases the presentation was asymmetric. One patient had autonomic dysfunction. The pathological findings in the biopsied sural nerves were those of axonal degeneration and vasculitis. In 6 nerves there was increased expression of Class II (Ia) antigen within the nerve fascicle, perineurium and within endothelial cells. PMID- 3032330 TI - Early effect of an ACTH4-9 analog (Org.2766) on regenerative sprouting demonstrated by the use of neurofilament-binding antibodies isolated from a serum raised by alpha-MSH immunization. AB - Antibodies binding to the 150 kDa neurofilament protein NF150 have been purified from a serum raised by immunizing a rabbit with alpha-MSH. The NF150-binding antibodies were purified by affinity chromatography on a column of cytoskeletal proteins coupled to CNBr-activated Sepharose-4B. Immunocytochemical application of these antibodies, followed by a FITC-coupled second antibody, labels axons in intact and regenerating nerves and provides a means of identifying and counting small regenerating sprouts from 48 h after nerve crush. We have been able to demonstrate that treatment with the neurotrophic melanocortin analog, Org.2766, increases the number of regenerating axons present in the nerve as early as 72 h after nerve crush. PMID- 3032331 TI - Dissociation of the IPSP and response to GABA during spreading depression-like depolarizations in hippocampal slices. AB - We have compared the sensitivity of CA1 and CA3 hippocampal pyramidal cells, in mature and immature tissue, to spreading depression-like depolarization episodes. Using hippocampal slices from rabbit, we have found that mature and immature tissue, and CA1 and CA3 neurons, were differentially prone to depolarization episodes, depending on the method used to produce the depolarization. CA1 region was generally more sensitive than CA3. Spontaneous and stimulus-evoked depolarizations were seen more frequently in immature tissue than in mature slices, but anoxia-induced depolarizations were much more likely to occur in mature tissue. Synaptic transmission and responses to somatic gamma-aminobutyric acid (GABA) ejection were compared during anoxia-induced depolarizations in mature slices. The early component of the inhibitory postsynaptic potential (IPSP) normally had the same reversal potential as the GABA response. During anoxia-induced depolarization, both the drug response and the PSPs were lost. Synaptic transmission generally disappeared before the response to exogenous GABA application; the GABA response reappeared before synaptic function was restored. During the recovery of resting potential (RMP) following depolarization, the reversal potential of the early IPSP differed significantly from that of the GABA response; when the cell had recovered to RMP, the IPSP was depolarizing, whereas GABA application produced a 'normal' cell hyperpolarization. IPSPs and GABA mediated responses attained their pre-depolarization form within a few minutes of RMP recovery. These observations suggest that, at least under special circumstances, the early component of the IPSP and GABA-mediated hyperpolarizations can be dissociated. Therefore, the early IPSP may be mediated by more complex mechanisms than a simple alteration in chloride conductance due to GABA-receptor interactions. PMID- 3032332 TI - Ventral pallidum projections to mediodorsal nucleus of the thalamus: an anatomical and electrophysiological investigation in the rat. AB - Horseradish peroxidase (HRP) and single unit recording experiments were done in rats to investigate neural connections from the ventral pallidal region to the mediodorsal nucleus of the thalamus (MD). In the first series, following the diffusion or iontophoretic injection of HRP into the MD, retrogradely labeled neurons were observed throughout the rostrocaudal extent of the ipsilateral ventral pallidum. Most of the labeled neurons were found in an area between the nucleus of the diagonal band and the ventral aspect of the substantia innominata subcommissuralis. Additional labeled neurons were found in the ventral aspect of the globus pallidus and substantia innominata sublenticularis. In the second series, the region shown to contain labeled neurons was explored for single units antidromically activated by single pulse stimulation of the MD in urethane anesthetized rats. One hundred and fifty-nine single units in the subpallidal area were antidromically activated with latencies corresponding to conduction velocities of 0.2-3.9 m/s. A greater percentage of units in the subcommissural region (50.3%) were activated antidromically as compared to the sublenticular region (27.4%). In the third series, the MD was explored for single units which responded orthodromically to stimulation of the ventral pallidum. Fifty-eight percent (40/69) of MD units responded to stimulation of the subcommissural substantia innominata, whereas 90% (72/80) MD units responded to stimulation of the sublenticular substantia innominata. The most frequent type of orthodromic response observed in MD neurons was inhibition with short onset latencies (less than 10 ms). These data provide anatomical and electrophysiological evidence for the existence of direct pathways from the ventral pallidum to the MD and suggest that this projection is part of a corticosubcortical loop through which the frontal cortex with the ventral striatum and pallidum may contribute to motor function. PMID- 3032333 TI - 'Non-specific' cholinesterase-containing neurons of the dorsal thalamus project to medial limbic cortex. AB - Thalamocortical neurons that contain 'non-specific' cholinesterase (ChE) were studied with cholinesterase histochemistry and experimental axonal tracing techniques in adult rats. In addition to the presence of ChE that is ubiquitous in capillary endothelium, neurons that contain ChE are found in 3 distinct regions of the dorsal thalamus, the thalamic reuniens nucleus (Re), the anterior dorsal nucleus (AD) and a region that includes the lateral part of the central lateral nucleus (CL) and the ventral portion of the lateral dorsal nucleus (LD). ChE activity appears light in cerebral cortex in general but histochemical staining is slightly greater in neuropil of the cingulate gyrus. Anterograde transport techniques with autoradiography demonstrated that neurons in the LD-CL region project to anterior cingulate cortex and the dorsal retrosplenial area. Anterograde degeneration techniques demonstrated that AD projects primarily to ventral retrosplenial cortex. Injections of horseradish peroxidase (HRP) in the anterior cingulate cortex resulted in double labeled cells (cells containing both ChE and HRP reaction products) primarily in LD and CL. HRP injections into ventral retrosplenial cortex resulted in double labeled cells in AD and Re. HRP injections in the subiculum resulted in double labeled cells in Re. Lesions placed in the region of thalamocortical projections resulted in a loss of ChE in the ipsilateral cingulate gyrus, as measured both histochemically and enzymatically. The finding that neurons containing ChE project to medial limbic cortex suggests that the ChE may be involved in the function of the thalamocortical component of the limbic system. PMID- 3032334 TI - Corticospinal projections originate from the arcuate premotor area. AB - We investigated the patterns of corticospinal projection to different segments of the cervical spinal cord using retrograde transport of wheat germ agglutinin conjugated to horseradish peroxidase. We confirmed prior suggestions that the arcuate premotor area (APA) has a significant projection to the spinal cord. In addition, we observed that the corticospinal projection from the APA terminates in upper, but not lower segments of the cervical spinal cord. Furthermore, the region of the APA which projects to the cervical cord also projects to the arm area of the primary motor cortex on the crest of the precentral gyrus. Thus, we conclude that the APA projection to upper cervical segments is involved in the cortical control of arm movements. PMID- 3032335 TI - Deafferentation elicits a transient decrease in Na+, K+-ATPase activity and ouabain binding in the olfactory tubercle. AB - This work examines the effects of olfactory bulbectomy on Na+, K+-ATPase activity and ouabain binding in the olfactory tubercle. The activity and number of enzyme sites were reduced significantly in olfactory tubercle, but not in corpus striatum or hippocampus, 14 and 21 days after bulbectomy. Enzyme activity and ouabain binding returned to normal by 42 days after the lesions. The time of the reduction in Na+, K+-ATPase coincides with that observed earlier for dopaminergic sprouting and increased dopamine-sensitive adenylate cyclase activity. PMID- 3032336 TI - The ventral striatopallidothalamic projection. III. Striatal cells of the olfactory tubercle establish direct synaptic contact with ventral pallidal cells projecting to mediodorsal thalamus. AB - After injection of HRP in the thalamic mediodorsal nucleus (MD) of the rat on one side followed by a superficial laminar heat lesion of the olfactory tubercle (Tu) on the same side, electron microscopic examination of DAB-reacted sections through the Tu revealed degenerating terminals contacting HRP-labeled neurons, confirming that a monosynaptic, two-neuron link extends, via a relay in ventral pallidum, between the superficial dense cell layer of the Tu and the MD. PMID- 3032337 TI - Sites of origin of gonadotropin releasing hormone containing projections to the amygdala and the interpeduncular nucleus. AB - The sites of origin of gonadotropin releasing hormone (GnRH) containing projections to the amygdala and the interpeduncular nucleus were studied with immunofluorescence for GnRH in combination with retrograde transport of True blue. After small injections of True blue into the amygdala, retrogradely labeled GnRH producing neurons were identified in the rostral medial septum, the caudal roots of the nervus terminalis, diagonal band, nucleus triangularis septi, nucleus interstitialis striae terminalis, and in the ventrolateral hypothalamus. After injection of True blue into the interpeduncular nucleus, a small GnRH producing cell group in the ventromedial diagonal band, and certain GnRH-positive neurons in the ventral hypothalamus had retrogradely transported the dye. The results suggest a link between the amygdala and the nervus terminalis which is provided in part by GnRH-containing projections. The heterogeneity of the sites of origin of inputs to the amygdala and the interpeduncular nucleus indicates that the 'behavioral' component of the GnRH system is not restricted to a single brain nucleus, but it may be part of a diffuse network which has close anatomical ties with the 'endocrine' GnRH system. PMID- 3032338 TI - 4-Aminopyridine affects synaptosomal protein phosphorylation in rat hippocampal slices. AB - Rat brain hippocampal slices were incubated with or without the convulsant 4 aminopyridine (4-AP). From these slices a crude mitochondrial/synaptosomal membrane fraction was prepared and analyzed for endogenous protein phosphorylation. 4-AP (10(-5) M) stimulated the phosphorylation of a 50 kDa protein by 86%. The phosphorylation of this 50 kDa protein is Ca2+/calmodulin dependent and we suggest that this protein is the lower molecular weight subunit of Ca2+/calmodulin-dependent protein kinase II (CaMK II). PMID- 3032339 TI - Pregnenolone-sulfate: an endogenous antagonist of the gamma-aminobutyric acid receptor complex in brain? AB - The interaction of the 'neurosteroid', pregnenolone-sulfate (PS), with the GABA/benzodiazepine/chloride ionophore receptor complex was investigated in rat brain subcellular preparations. At low micromolar concentrations PS competitively inhibited the binding of the convulsant [35S]t-butylbicyclophosphorothionate (TBPS) and antagonized pentobarbital-stimulated [3H]flunitrazepam binding to synaptosomes. In addition, PS inhibited muscimol-stimulated 36Cl-uptake in brain synaptoneurosomes, including that PS has characteristics of a relatively potent antagonist of the chloride channel coupled to the GABA receptor. Together with our previous finding that A-ring reduced metabolites of progesterone and deoxycorticosterone also interact with the GABA receptor complex but as hypnotic barbiturates, these data suggest that the regulation of GABAergic neurotransmission by various neurosteroids may be an important mechanism for controlling neuronal excitability. PMID- 3032340 TI - Functional and metabolic correlates of long series of cortical spreading depression waves in rats. AB - Repetitive generation of spreading depression (SD) waves may induce metabolic and functional disturbances in the invaded brain regions. In order to better characterize this state, up to 60 SD waves were elicited in the cerebral cortex of anesthetized rats by intracortical injections of 2 microliters of isotonic K acetate repeated at regular 5-min intervals. Propagation rate of SD between capillary electrodes located 1 mm and 7 mm from the injection site gradually decreased and after 35-45 waves became irregular. In another series of experiments SD-induced changes of cyclic adenosine monophosphate (cAMP) level were examined after 24 SD waves. Although no irregularities of SD propagation were observed, maximum level of cAMP in the SD-invaded cortex decreased from 270% in controls to 170% after 24 waves. Also, the recovery of the nucleotide was slowed down. Metabolic alterations thus preceded irregularities in SD propagation. PMID- 3032341 TI - Long C3-C5 propriospinal neurones in the cat. AB - Intracellular recording was made in the C3-C5 segments of cats from cells identified as long propriospinal neurones (PNs) by antidromic activation from the lower thoracic segments. The cell bodies were in laminae VII and VIII and their ventrally located axons were either uncrossed or crossed. Stimulation of higher motor centres revealed monosynaptic excitatory postsynaptic potentials (EPSPs) from cortico-, rubro-, tecto-, reticulo-, interstitio-, fastigio- and trigeminospinal fibres. Monosynaptic inhibitory postsynaptic potentials (IPSPs) were evoked from reticulospinal fibres. These PSPs were in addition to the separately described effects from the vestibular nuclei. Monosynaptic EPSPs were also evoked in some cells from neck or forelimb afferents and disynaptic EPSPs or IPSPs from forelimb afferents. PMID- 3032342 TI - Noradrenergic regulation of cytosol estrogen receptors in female rat hypothalamus: possible role of alpha 2-noradrenergic receptors. AB - Because of the previous work demonstrating that the alpha 1-noradrenergic receptor antagonist, prazosin, decreases the concentration of cytosol estrogen receptors in rat mediobasal hypothalamus, a series of experiments was performed to determine the specificity of this effect to the alpha 1-noradrenergic system. Injection of the alpha 2-noradrenergic antagonist, yohimbine, caused a decrease in the concentration of estrogen receptors in mediobasal hypothalamus. In addition, the down-regulation of cytosol estrogen receptors by either the alpha 1 noradrenergic antagonist, prazosin, or the alpha 2-noradrenergic antagonist, yohimbine, could be blocked by pretreatment with the alpha 2-noradrenergic agonist, clonidine. The alpha 1-noradrenergic agonist, phenylephrine, was ineffective in blocking the effects of the alpha 1-noradrenergic antagonist, prazosin. These results add further support to the hypothesis that the alpha noradrenergic system modulates the concentration of cytosol estrogen receptors in the rat hypothalamus. They suggest that the modulation may occur by way of alpha 2-noradrenergic receptors in addition to, or instead of, alpha 1-noradrenergic receptors. PMID- 3032343 TI - Beta-adrenergic receptors of cerebellar astrocytes in culture: intact cells versus membrane preparation. AB - Experiments were carried out to assess: the influence of culture conditions on the expression of beta-adrenergic receptors in intact glial cells from the central nervous system; and the extent to which quantitation of receptor sites in membrane preparations reflects the receptor population of the whole cells they are derived from. Cerebellar astrocytes were chosen for this study since essentially one receptor subtype, the beta 2 one, is present in adult cerebellum. Intact, attached cerebellar astrocytes exhibit only one class of binding sites for the beta-adrenergic antagonist, [3H]CGP 12177. Replating of the astrocytes after a few days of culture in vitro induces an up-regulation of the receptors. This effect is particularly important when astrocytes are maintained for 6 days in the presence of horse serum, a condition that favors cellular differentiation. Only 30-50% of the beta-adrenergic receptors of the intact cells can be detected on membrane preparations. When membranes are prepared from astrocytes grown either in the presence of horse serum or under chemically controlled medium (i.e. under differentiation promoting conditions) two classes of binding sites for [125I](-)-iodocyanopindolol are revealed. Several hypotheses, mainly related to the morphology of the cells, may provide an explanation for such differences. Studies of the pharmacological specificity of receptors of membrane fractions show that cerebellar astrocytes cultured in vitro exhibit both beta 1 and beta 2 receptor subtypes. The beta 1 subtype receptors are slightly more abundant when astrocytes are grown in fetal calf serum (FCS), a condition under which they exhibit a polygonal, poorly differentiated morphology. When culture conditions favor cellular differentiation, more receptors of the beta 2 subtype are seen, which can be related to what is observed in the adult in vivo where the astrocytes exhibit a differentiated morphology. PMID- 3032344 TI - Alpha 2-adrenoceptors in amygdala control renal sympathetic nerve activity and renal function in conscious spontaneously hypertensive rats. AB - The contributions of alpha 2- and beta 2-adrenoceptors in the central amygdaloid nucleus to the increased renal sympathetic nerve activity and decreased urinary sodium excretion resulting from environmental stress (air jet) in conscious spontaneously hypertensive rats (SHR) were examined. Air stress increased mean arterial pressure and renal sympathetic nerve activity (47% from 9.3 +/- 0.8 integrator resets/min), and decreased urinary sodium excretion (38% from 2.4 +/- 0.3 microEq/min/100 g b. wt.). After bilateral administration of guanabenz (2 X 2.5 micrograms) (alpha 2-adrenoceptor agonist) into the central amygdaloid nucleus of the same SHR, air stress had no effect on renal sympathetic nerve activity (+7% from 7.5 +/- 0.8 integrator resets/min) or urinary sodium excretion (+5% from 3.5 +/- 0.4 microEq/min/100 g b. wt.), but still increased mean arterial pressure. These effects of guanabenz were prevented by the prior bilateral administration of the alpha 2-adrenoceptor antagonist, rauwolscine (2 X 15 micrograms), into the central amygdaloid nucleus; rauwolscine alone had no effect on the renal responses to air stress in SHR. Guanabenz in the lateral cerebral ventricle (2 micrograms), the basolateral amygdaloid nucleus (2 X 2.5 micrograms) or a site dorsal to the central amygdaloid nucleus in the area of globus pallidus (2 X 2.5 micrograms) had no effect on the renal sympathetic nerve activity or urinary sodium excretion responses to air stress. Similarly, ICI 118,551 (2 X 1 microgram; beta 2-adrenoceptor antagonist) in the central amygdaloid nucleus had no effect on the renal responses to air stress.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3032345 TI - Cross-tolerance between analgesic low doses of morphine and naloxone in arthritic rats. AB - The effects of acute injections of naloxone (3-3000 micrograms/kg i.v.) and morphine (100-1000 micrograms/kg i.v.) on the vocalization threshold induced by pressure on the paw were analyzed in adjuvant-induced arthritic rats pretreated either with naloxone or with morphine administered at low doses (9 micrograms/kg s.c. and 3000 micrograms/kg s.c., respectively) over 4 consecutive days. In naloxone-pretreated arthritic rats, the paradoxical analgesic effect of low doses of naloxone was almost abolished, and the potent analgesic effects of low doses of morphine were also strongly and dose-dependently reduced. In morphine pretreated arthritic animals, the analgesic effect of low doses of naloxone was significantly attenuated. These results attest that a cross-tolerance with low analgesic doses of morphine and naloxone can be demonstrated in these chronic suffering animals. By contrast, in rats pretreated either with naloxone or morphine, the hyperalgesic effect of naloxone produced by higher doses persisted and even was unmasked for doses which were analgesic before the pretreatment. These data emphasize the involvement of opiate receptors different in their sensitivity and/or their functions in the two opposite effects of naloxone. They also suggest that opiate receptors and endorphinergic systems differ in normal animals and animals which experience persistent pain. PMID- 3032346 TI - Phaclofen: a peripheral and central baclofen antagonist. AB - Phaclofen, the phosphonic acid derivative of baclofen, reversibly antagonized the depression of the cholinergic twitch response of the guinea pig ileum and distal colon by either baclofen or GABA. When administered microelectrophoretically, phaclofen reversibly blocked the presumed presynaptic reduction by baclofen of the monosynaptic excitation of spinal interneurones by impulses in primary afferent fibres of the cat but did not block the postsynaptic depressant action of baclofen on these neurones. Phaclofen may thus be useful in determining the physiological significance of central and peripheral bicuculline-insensitive receptors with which GABA and (-)-baclofen interact. PMID- 3032347 TI - The modulation of dopamine-sensitive adenylate cyclase activity in the mouse caudate nucleus by estradiol. AB - When levodopa (L-DOPA) is administered to mice, the central nervous system responses to the dopamine stimulation were less sensitive in female than in the male mice. The sensitivity is reflected both behaviorally and biochemically. The behavioral response were measured by scores recorded after intraperitoneal injection of a standard dose of L-DOPA. The biochemical studies included the determination of the cyclic AMP produced by in vitro experiments after incubating dopamine with mouse caudate nucleus homogenates. This insensitivity to dopamine stimulation in female animals can be reversed by ovariectomy. Estradiol treatment in vivo suppressed the dopamine stimulation, while estradiol directly added to the caudate nucleus homogenate does not affect the dopamine-sensitive adenylate cyclase activity. These findings indicate that the estradiol-induced effects on the brain enzyme are indirect. The female sex hormone does have an effect on the central dopaminergic system. However, the underlying mechanism(s) cannot be delineated by the present studies. PMID- 3032348 TI - Facilitation of premotor cortical seizure development by intranigral muscimol. AB - The role of the substantia nigra in seizure development was investigated using a chronic model of partial onset generalized seizure induced by low frequency cortical stimulation. Unilateral intranigral micro-injection of muscimol, a GABA receptor agonist, was found to facilitate partial onset seizure development, but did not affect developed seizures. This finding suggested that a non dopaminergic, presumably GABAergic, mechanism was involved since the facilitatory effect of intranigral muscimol was not modified by haloperidol pretreatment. PMID- 3032349 TI - Intercostal muscles and purring in the cat: the influence of afferent inputs. AB - Feline purring has previously been reported as originating in a central oscillator, independent of afferent inputs, and also as not involving expiratory muscles. Here we show, via electromyographic recordings from intercostal muscles, quantified by cross-correlation, that expiratory muscles can be involved and that even if the oscillator is central, reflex components nevertheless play a considerable part in the production of the periodic pattern of muscle activation seen during purring. PMID- 3032350 TI - Activity of A9 and A10 dopaminergic neurons in unrestrained rats: further characterization and effects of apomorphine and cholecystokinin. AB - The activity of single dopaminergic (DA) neurons (total n = 77) in the midbrain of awake, unrestrained rats was examined. The firing rates and patterns of ventral tegmental area (A10) cells (n = 39) were similar to those of identified DA neurons in anesthetized and paralyzed rats. Unit activity was briefly stimulated (increased firing rate and burst activity) in response to an auditory stimulus and during manual stimulation of the vibrissae. Similar changes occurred during orienting responses and periods of sniffing. Twenty-six percent of A10 cells recorded appeared to be electronically coupled which matched the prevalence previously observed among A9 neurons. The effects on A9 and A10 DA cell activity of apomorphine and the carboxyterminal octapeptide of cholecystokinin (CCK-8) were then determined. Sequential doses of apomorphine (5-320 micrograms/kg, i.v.) reduced the firing rate of each neuron tested (n = 19). Ten of these cells were classified as 'sensitive' to the drug (ED50 less than 20 micrograms/kg), while the remainder were considerably 'less sensitive' (ED50 greater than 30 micrograms/kg). Cells of either sensitivity were as likely to be found in A9 as in A10. Sulfated CCK-8 (1-16 micrograms/kg, i.v.) excited (firing rate and bursting increased) 73% of the A9 neurons sampled but produced inconsistent effects on A10 cell firing. CCK-8 pretreatment increased the percentage of A9 and A10 neurons which were classified as 'sensitive' to apomorphine (82%). Enhanced sensitivity to apomorphine occurred regardless of the effect of CCK-8 on unit firing. Thus, as has been found in anesthetized rats, CCK-8 appeared to enhance the inhibitory effects of a DA agonist on DA neurons. PMID- 3032351 TI - Intracellular recordings in pericruciate neurons during spike and wave discharges of feline generalized penicillin epilepsy. AB - Concurrent EEG and intracellular recordings from pericruciate neurons of cats obtained before and after i.m. injection of penicillin inducing the syndrome of feline generalized penicillin epilepsy (FGPE) characterized by spike and wave (SW) discharge in the EEG, display large excitatory postsynaptic potentials (EPSPs) at the time of the EEG 'spike' which alternate with hyperpolarizing potentials occurring in coincidence with the EEG 'wave' component of the SW complex. The large EPSPs trigger discharges of single or multiple high-frequency action potentials which do not show a progressive decrement in amplitude nor an appreciable increase in duration. These bursts thus differ in some respects from typical paroxysmal depolarization shifts. The hyperpolarizing potentials show an early phase which is reversed by intracellular Cl- injection or diffusion and thus behaves like a classical inhibitory postsynaptic potential (IPSP). The late phase is unaffected by Cl-. Hyperpolarizing potentials of pericruciate neurons induced by antidromic activation of the cerebral peduncle (CP) or by direct cortical stimulation are not altered after i.m. injections of penicillin at doses sufficient to induce generalized SW discharge. The early phase of hyperpolarization both before and after i.m. penicillin is reversed by intracellular Cl- injection or diffusion, the late phase remains unchanged. The early phase thus represents a classical IPSP, which does not appear to be affected by the low brain penicillin concentrations sufficient to induce generalized SW discharge. It is concluded that this form of epileptic discharge cannot be attributed to blockage of phasic (presumably somatic) postsynaptic inhibition by penicillin. These results indicate that to regard all forms of epileptic discharge as the consequence of a blockage of gamma-aminobutyric acid mediated phasic postsynaptic inhibition acting on the soma represents an unduly restrictive view of epileptogenesis. PMID- 3032352 TI - Time course study on the effect of reserpine on hypothalamic immunoreactive CRF levels in rats. AB - A time course study on the changes of rat hypothalamic corticotropin-releasing factor (CRF) levels and ACTH levels in plasma, pituitary and hypothalamus after an acute treatment with reserpine was examined using a rat CRF RIA. The massive and prolonged depletion of hypothalamic norepinephrine and dopamine levels provoked by a single injection of reserpine (2 and 8 mg/kg, i.p.) caused a transient decrease of hypothalamic CRF levels and ACTH levels in the anterior pituitary glands, and an increase in plasma ACTH levels. There was a strong correlation between the depletion of hypothalamic CRF and norepinephrine levels. These results suggest that: acute depletion of hypothalamic norepinephrine levels cause the initial release of CRF that stimulates pituitary ACTH secretion, and the depletion of CRF and ACTH stores at the early stage; and noradrenergic pathways may be involved in the inhibitory mechanism of CRF release. PMID- 3032353 TI - Naloxone-reversible analgesia induced by electrical stimulation of the habenula in the rat. AB - During a previous study of the nucleus parafascicularis (Pf), cells were recorded in the lateral habenula (HbL) which exhibited response patterns to peripheral noxious stimuli similar to those recorded in the Pf. In order to study the possible role of the habenular complex (Hb) in pain processing, we investigated the effect of electrical stimulation of the Hb on the tail-flick latency. For each series of experiments, the Hb of 15 female rats was implanted unilaterally, with bipolar electrodes, on either the right or left side. A week later, the animals were submitted to measurements of tail-flick latency, every 10 min, for a period of 3 h. The amount of analgesia was estimated by the percentage increase in latency. Five intensities of current (50, 100, 200, 300 and 400 microA) were used for stimulation during 60 s, at 50 Hz and 0.5-ms pulse width. A group of animals were given naloxone i.p. (1 mg/kg) 40 min after Hb stimulation at 200 microA to study the reversibility of the analgesia. A second group had their Hb destroyed by coagulation and the effect on tail-flick latency was checked once a week for 4 weeks. The results of these experiments clearly demonstrate Hb stimulation-induced analgesia, the maximum of which occurs 60-80 min after stimulation and then decreases slowly. The maximal amount of analgesia increases with the intensity of current up to 200 microA, without any behavioral side effects. At 300 microA, the analgesia is not significantly different from the one induced at 200 microA. However at 400 microA, behavioral side effects (fear, escape) appear and the analgesia is weaker. Two-hundred microA appears to be the most efficient current intensity and induces an average of 80% increase in tail flick latency. The group which was given naloxone exhibited a dramatic and complete reversal of analgesia. The group which had their Hb destroyed did not show any difference from the control group a week after surgery. During the following weeks, both lesioned animals and controls exhibited a habituation-like analgesia, without any significant difference (the index of analgesia was 45.73 +/- 23.65% for the lesioned rats and 51.82 +/- 29.18% for the controls), which was not naloxone reversible. A review of the literature does not provide an explantation for Hb-induced analgesia.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3032354 TI - Inhibition of synaptic transmission in the hippocampus by cholecystokinin (CCK) and its antagonism by a CCK analog (CCK27-33). AB - The actions of CCK-8 and putative CCK-8 antagonists were examined on synaptic transmission in the hippocampal slice preparation. CCK-8 (10(-7) M) caused a reversible depression of postsynaptic responses of the Schaffer collateral-CA1 synapse. CCK-mediated depression was unaffected by proglumide (10(-3) M) but was antagonized by a CCK analog (CCK27-33). The CCK analog alone had an excitatory effect on synaptic transmission. These results suggest that CCK has an inhibitory action on synaptic transmission. PMID- 3032356 TI - Selective changes in mu, delta and kappa opioid receptor binding in certain limbic regions of the brain in Alzheimer's disease patients. AB - Total opioid binding and levels of the three major types of opioid binding sites were measured in homogenates of various limbic structures from post-mortem brains of Alzheimer's disease patients and age-matched control individuals. The most consistent finding in Alzheimer's disease brains was an increase in kappa binding in all 6 areas of the limbic system examined, with the putamen and caudate regions showing significant increases of 114% and 53%, respectively. In addition, the Alzheimer's disease putamen showed a significantly higher level of total binding (85% increase). The amygdala of Alzheimer's disease patients exhibited significantly lower levels of mu and delta binding (41% and 55% decrease, respectively). Total binding and binding to mu and delta receptors in frontal cortex, caudate and hippocampus of Alzheimer's disease brains was indistinguishable from levels seen in these brain areas from control individuals. PMID- 3032355 TI - Quantitative histochemical and microchemical changes in the adult mouse central nervous system after section of the infraorbital and optic nerves. AB - The sensory projections from the whiskers of mice and other rodents synapse somatotopically in 3 subnuclei in the brainstem trigeminal complex, in the ventrobasal complex of the thalamus and in the somatosensory cortex. Deafferentation of the whiskers in adult animals results in qualitative and quantitative changes in activities of the metabolic enzymes in the somatosensory cortex (e.g. J. Neuro-sci., 1 (1981) 929-935). We determined the time course and extent of changes in the subcortical trigeminal centers of adult mice after deafferentation. The right infraorbital nerve was sectioned in mice under surgical anesthesia; the animals survived for periods up to 26 weeks. The optic nerve was also cut to evaluate the effects of central tract section. Some brains were prepared histochemically for the mitochondrial enzymes cytochrome oxidase (CO) and succinic dehydrogenase (SDH), and some were prepared for microchemical analysis of the enzymes citrate synthase (CS), malate dehydrogenase (MDH) and phosphorylase. All deafferented and intact nuclei were examined in each animal quantitatively. The oxidative enzymes (CO, SDH, CS and MDH) that were analyzed by histochemical and microchemical approaches showed a decrease in activities as early as 3 weeks postdeafferentation, a trend that continued up to 12 weeks in all the subcortical trigeminal stations and lateral geniculate nucleus (LGN) when compared with the intact side. By 25 weeks postlesion, the levels were comparable to the intact side except that in the LGN, the levels remained depressed. The phosphorylase levels increased at around 3 weeks postoperation and remained elevated 25 weeks postlesion. Each case provided results on the effects of deafferentation at a given time point throughout the trigeminal pathway. Direct quantitative correlation of histochemical and microchemical approaches for glycolytic enzymes is consistent with a coordinate regulation of these molecules. The changes in enzyme levels in all nuclei occur simultaneously and to a similar degree. This strongly suggests that neuronal activity plays an important role in regulating metabolic machinery throughout this pathway in adults. PMID- 3032358 TI - Direct excitatory and lateral inhibitory synaptic inputs to amacrine cells in the tiger salamander retina. AB - Two distinct synaptic currents in amacrine cells were measured using whole cell patch clamp in retinal slices. Synaptic activity was elicited with transretinal current passed from photoreceptors to ganglion cells to depolarize bipolar cell terminals. Recordings made in line with the stimulating electrodes revealed inward synaptic currents that reversed near O mV, positive to spike threshold. Recording 400 micron lateral to the stimulating electrodes revealed synaptic currents that reversed near-60 mV, negative to spike threshold. The lateral signal was blocked by tetrodotoxin and strychnine. A current with reversal potential similar to the lateral input was elicited by direct application of glycine to the amacrine cells. The results suggest that bipolar, but not amacrine input can elicit spikes in neighboring amacrine cells, that amacrine cells are mutually inhibitory and the inhibitory lateral transmission is limited to the extent of processes of the excited, spiking cell. PMID- 3032357 TI - Release of cholecystokinin from rat cerebral cortex in vivo: role of GABA and glutamate receptor systems. AB - Using cortical cups in chloralose-urethanized rats, the in vivo release of cholecystokinin-like immunoreactivity (CCK-LI) from cerebral cortex was examined. Resting levels of cholecystokinin-like immunoreactivity ranged from 20 to 30 pg/20 min sample. The addition of potassium (40 mM) in excess, resulted in a highly significant elevation in the levels of CCK-LI in the cortical superfusate. Deletion of calcium and the substitution of cobalt (10 mM), resulted in a significant reduction in both resting release and the release otherwise evoked by the addition of potassium. Focal electrical stimulation of the cortex (20 Hz), resulted in a significant (1.9 +/- 0.2-fold, n = 8) increase in the levels of CCK LI. The addition of glutamate (10(-6)-10(-4) M) of kainic acid (10(-8)-10(-6) M), also resulted in significant elevations in the levels of CCK-LI. The co administration of a putative glutamate receptor antagonist, kynurenic acid (10( 4) M) resulted in a significant reduction in the levels of release otherwise evoked by the addition of glutamate, but not by electrical stimulation. The addition of GABA (10(-5)-10(-3) M) resulted in a dose-dependent decrease in the resting release of CCK-LI, and the release evoked by glutamate. Picrotoxin (10( 6)-10(-4) M), resulted in a highly significant increase in the levels of CCK-LI in the cortical effluent. These results are consistent with a tonic GABAergic inhibition of CCK-releasing neurons. The treatment of the animal with diazepam (30 mg/kg, i.p.) also resulted in a significant reduction in resting release and the release otherwise evoked by focal cortical stimulation. PMID- 3032359 TI - Vasoconstriction and neural excitation in response to transient hypoxia in the rat hippocampal slice. AB - The vascular and neural responses to transient hypoxia in the rat hippocampal slice were studied. Neural hyperexcitability produced by tissue hypoxia was associated with localized decreases in the diameter of precapillary arterioles. Vasoconstriction occurred periodically along the length of vessels observed. The mean percent decrease in vessel diameter in these narrowed regions was 10.25%. Population spikes recorded in the cell body layer of the dentate gyrus showed a mean increase in amplitude of 71.3%. The mean latency to peak response was similar for both the vessels and neurons. The results suggest mechanisms by which autoregulatory influences on microvessel caliber may be counteracted in conditions of hypoxia and hypotension in the whole animal. PMID- 3032360 TI - Projection from the prefrontal cortex to histaminergic cell groups in the posterior hypothalamic region of the rat. Anterograde tracing with Phaseolus vulgaris leucoagglutinin combined with immunocytochemistry of histidine decarboxylase. AB - We investigated the projection from the infralimbic division of the prefrontal cortex (area 25) to histaminergic neurons in the posterior hypothalamic area. Phaseolus vulgaris-leucoagglutinin (PHA-L) was injected in the prefrontal cortex of rats. Frozen brain sections were subjected to combined PHA-L and histidine decarboxylase (HDC)-peroxidase immunocytochemistry, using nickel-enhanced diaminobenzidine (blue reaction product) to visualize the transported PHA-L, and diaminobenzidine (brown reaction product) to visualize simultaneously the HDC containing neurons. PHA-L-labeled fibers could be seen coursing in the capsula interna, leaving the telencephalon via the anterior thalamic radiation and the medial forebrain bundle. In the lateral and posterior hypothalamic areas, PHA-L labeled fibers leave the medial forebrain bundle and traverse the nuclei containing HDC-immunoreactive neurons. Varicosities on the PHA-L-labeled fibers, the majority of which occur en passant, could be observed in close association with the HDC-immunoreactive neurons. The results suggest that the hypothalamic histaminergic neurons receive afferent synaptic input from neurons of the infralimbic division of the prefrontal cortex. PMID- 3032361 TI - Responses of adrenocorticotropin and vasopressin to hemorrhage after lesions of the caudal ventrolateral medulla in rats. AB - Hormonal responses to hemorrhage were examined in urethane-anesthetized rats 48 h after electrolytic lesions of the caudal ventrolateral medulla. Bilateral A1 lesions (A1X) inhibited the response of plasma adrenocorticotropin relative either to other lesions (XX) or to sham lesions. A1X inhibited the response of plasma vasopressin relative to XX but not to sham lesions. A1X and XX destroyed similar areas that were dorsal or caudal to A1. No lesion blocked either response completely. A hemodynamic pathway controlling the release of both hormones may pass through A1. However, other pathways may bypass this area. PMID- 3032362 TI - Pertussis toxin or 8-bromo-cAMP block inhibition of the acoustic startle response by the alpha 2-adrenergic agonist ST-91. AB - Stimulation of supraspinal alpha 2-adrenergic receptors by intraventricular infusion of the alpha 2-adrenergic agonist ST-91 depresses a simple vertebrate behavior, the acoustic startle response. Intraventricular pretreatment with pertussis toxin, an agent known to inactivate the inhibitory guanine nucleotide binding protein (Gi) which can inhibit adenylate cyclase, completely prevented the depressant behavioral effect of ST-91. In contrast, pertussis toxin did not alter the depressant effect of intraventricular infusion of the 5-HT 1B agonist 1 m-chlorophenylpiperazine (mCPP). Intraventricular infusion of the cyclic adenosine monophosphate (cAMP) analog 8-bromo-cAMP also reversed the depressant effect of ST-91 without altering the effect of mCPP. These data suggest that inhibition of adenylate cyclase may be involved in the effect of activation of central alpha 2-adrenergic receptors. PMID- 3032363 TI - Barbiturate-induced inhibition of a spinal nociceptive reflex: role of GABA mechanisms and descending modulation. AB - The present study investigated the effect of systemically administered pentobarbital on the tail-flick (TF) reflex in rats, the neurochemical mechanism of action and the role of descending influences. Pentobarbital produced a clear inhibition of the TF response. Systemic administration of naloxone did not significantly alter this effect, thus it appears to be independent of endogenous opioid systems. Complete spinal transection resulted in a marked potentiation of pentobarbital-induced TF inhibition, demonstrating a spinal locus of action. Moreover, this observation suggests the existence of a tonic descending excitatory influence, opposing the pentobarbital-produced depression of nociceptive transmission in the intact animal. Intrathecal administration of pentobarbital caused a much more pronounced TF inhibition in transected than in intact animals, lending further support to this hypothesis. To identify the neurochemical mechanisms involved in pentobarbital-produced antinociception, the gamma-aminobutyric acid (GABA) antagonists bicuculline and picrotoxinin were administered intrathecally in spinalized animals. Both substances caused an attenuation of the pentobarbital effect, demonstrating the involvement of GABAergic transmission. The proposed descending excitatory system may act either presynaptically and cause a decreased release of GABA into the synapse or postsynaptically via endogenous GABA antagonistic neurotransmitters, which may change the conformation of the GABA-barbiturate receptor complex. PMID- 3032364 TI - Chronic lithium treatment increases the phosphorylation of a 64-kDa protein in rat brains. AB - Despite the wide clinical use of lithium in the treatment of manic depressive illness there is no adequate explanation for its mechanism of action. In the light of lithium's suggestive effects on the second messenger system in the brain, we studied the effects of chronic dietary lithium treatment (achieving blood levels in the therapeutic range) on protein phosphorylation in different areas of rat brain. An increase in the phosphorylation of a 64-kDa membrane associated protein was evident in the lithium-treated rats compared to controls. This increase was observed only under basal phosphorylating conditions and was abolished when the phosphorylation was performed in the presence of Ca2+ or Ca2+ and calmodulin. The possibility that this 64-kDa protein affected by lithium is the beta-subunit of the calmodulin-dependent protein kinase or a different protein which co-migrates with it is discussed. PMID- 3032365 TI - Role of cholinergic neuromuscular transmission in the neuroregulation of the autophosphorylatable regulatory subunit of cyclic AMP-dependent protein kinase type II and the acetylcholine receptor content in skeletal muscle. AB - Previously, we found that the in vitro [32P]-autophosphorylation of the regulatory subunit of cyclic AMP-dependent protein kinase type II in rat soleus muscles is subject to a nerve-stump-length-dependent neuroregulation which indicates that this event is dependent upon some neural signal other than the impulse-directed release of acetylcholine. In this investigation, tetrodotoxin and alpha-bungarotoxin were also administered to further differentiate the effect of impulse-directed and spontaneously released acetylcholine upon this event and also upon the appearance of new acetylcholine receptors as measured by the binding of radioiodinated bungarotoxin. A 24 h blockade of cholinergic transmission with either neurotoxin did not change the phosphorylation level of the regulatory subunit, while a significant increase is observed when solei are surgically denervated for this period. The phosphorylation level and also the acetylcholine receptor content were increased only after more prolonged (48-96 h) muscle inactivity was produced with the neurotoxins. However, then their effects may not be solely related to alterations in cholinergic transmission. Taken together, our results do not support a trophic role for spontaneously released acetylcholine with respect to the two neurotrophic events studied. PMID- 3032366 TI - Dopamine agonists produce functional recovery from septal lesions which affect hypothalamic defensive attack in cats. AB - Effects of lesions of the lateral septum and subsequent administration of methamphetamine (MAT, 1 mg/kg, i.p.) or apomorphine (APO, 1 mg/kg, i.p.) on thresholds for defensive attack elicited by electrical stimulation of the ventromedial hypothalamic nucleus (VM) were examined. Hissing and directed attack were selected for threshold determination. Thresholds were measured under two situations: one with provocation by a human and the other without it. Electrolytic lesions of the lateral septum enhanced the facilitative influences exerted by the provocation on the thresholds, however, subsequent administration of MAT or APO abolished or tended to abolish the enhancement. The rapid recovery of function was interpreted to have taken place due to excessive dopaminergic inputs to the spared tissue of the lateral septum, and a gating mechanism of neuronal information by dopamine was suggested. PMID- 3032367 TI - Heat production associated with synaptic transmission in the bullfrog spinal cord. AB - By using heat detectors made with pyroelectric film, rapid heat production by the bullfrog spinal cord in response to dorsal root stimulation has been demonstrated. The heat production rises to its peak in about 100 ms after the arrival of afferent impulses and falls slowly with a time course comparable to that of the dorsal root potential. Stimulation of the ventral roots produces no detectable heat. The heat production was reversibly suppressed by immersion of the cord in a low Ca2+, high Mg2+ salt solution, indicating that the underlying exothermic process is associated with intraspinal synaptic transmission. The source of this 'synaptic heat' is located near the boundary between the dorsal column and the substantia gelatinosa in the vicinity of the stimulated dorsal roots. PMID- 3032368 TI - Occurrence of the D-1 dopamine receptor in the substantia nigra of several mammalian species: identification in binding studies using [125I]SCH 23982. AB - The substantia nigra of the rat brain contains a high concentration of binding sites for benzazepine antagonists of the D-1 receptor. Using [125I]SCH 23982, the properties of these binding sites are characterized and shown to resemble the D-1 dopamine receptor occurring in the caudate nucleus. Similar binding sites are identified in the substantia nigra of the cat and guinea pig. PMID- 3032369 TI - Effects of neonatal thermal lesioning of the mesocortical dopaminergic projection on the development of the rat prefrontal cortex. AB - The role of dopamine (DA) in the development of the prefrontal cortex (PFC) was investigated by depleting the dopaminergic innervation of the PFC. A new stereotaxic procedure made it possible to produce small lesions in 1-day-old rats confined to the A10 group of dopaminergic cell bodies in the ventral tegmentum, from which the dopaminergic projection to the PFC originates. The variety in the lesions revealed a clear topographical organization of the efferent connections of the ventral tegmental area (VTA) to the prefrontal cortex. As far as we know from the literature the data presented in this study are a first direct indication of a neurotrophic role for dopamine in the development of the prefrontal cortex. When the prefrontal cortex was depleted of the dopaminergic innervation from birth on, by lesioning the cells of origin on postnatal day 1, the cortical thickness in the medial PFC was reduced by about 6%. Although coagulative lesions in the ventral tegmentum cause also a depletion of cortical serotonin, cortical reduction seems to be rather the result of the absence of dopamine during its development. This is indicated by the absence of a significant cortical thickness reduction in the dysgranular part of the first somatosensory cortex, which receives a serotonergic but no dopaminergic innervation. PMID- 3032371 TI - Noradrenergic regulation of alpha 1-receptors during the postnatal development of the guinea pig. AB - In guinea pig brain, alpha 1-noradrenergic receptor concentrations undergo region specific fluctuations during the first weeks of postnatal life. However, the factors involved in the regulation of these receptors have yet to be identified. In this study, the ontogeny of one possible regulatory factor, norepinephrine, was examined in relation to postnatal changes in alpha 1-receptor levels in several different regions of guinea pig brain. Results from these studies showed that while the activity of the noradrenergic system increased throughout the first weeks of postnatal development in each brain area examined, the concentration of alpha 1-receptors decreased in preoptic area and hypothalamus and increased in cortex. In subsequent experiments, the effects of noradrenergic lesions with 6-hydroxydopamine on alpha 1-receptor levels were assessed to examine the possibility that alpha 1-receptors are differentially sensitive to noradrenergic stimulation in cortex and preoptic area/hypothalamus in immature guinea pigs. Noradrenergic lesions which reduced norepinephrine levels by 87-94% resulted in significant elevations in alpha 1-receptors in all regions examined. These results are discussed with reference to the anatomical distribution of alpha 1-receptors and their regulation by norepinephrine. PMID- 3032370 TI - Exogenous cholecystokinin (CCK) reduces neonatal rat brain opioid receptor density and CCK levels. AB - Newborn rats were given saline or cholecystokinin8 (CCK8) (5 micrograms/kg, twice daily) i.p. for 3 weeks. On day 21, effects on brain development were assessed. CCK-like immunoreactivity was measured in 7 brain regions; a small (12-18%) but significant decrease in endogenous levels of this peptide was detected in cerebral cortex, medulla and pons of the CCK-treated rats. Morphometric measurements revealed a slight reduction in thickness of most cerebral cortical sections within the CCK-treated group. The area of a midsagittal section of the cerebellum was unchanged except for the Purkinje/granule cell layer, which was smaller in CCK-treated animals. Levels of mu-, delta- and kappa-opioid receptors were estimated by homologous displacement binding assays using selective radioligands. The CCK treatment resulted in a significant decrease in levels of mu- (11%) and delta- (13%)-sites in the cerebral cortex. Neither binding affinities nor kappa-receptor densities were altered. Other animals received the same treatment regimens for 21 days and were maintained for an additional 29 days without treatment; these rats had reductions only in cortical mu-sites (15%). Chronic intraventricular administration of CCK (0.1 microgram/h) to adult rats did not elicit a similar down-regulation of cortical mu or delta receptors, suggesting that the effects observed in neonates reflected developmental processes. PMID- 3032372 TI - Motoneurone survival and neurite regeneration requirements: the role of dorsal root ganglion cells during development. AB - The effect of dissociated dorsal root ganglion cells on the survival of motoneurones in vitro from differently aged chick embryos has been studied. Homogeneous cultures of motoneurones were prepared from 5-8-day embryos, using a cell sorter; dorsal root ganglion cells were obtained from 8-day embryos. The survival of motoneurones from 5-day and 6-day embryos was not enhanced above controls by the presence of dorsal root ganglion cells; however, the survival of motoneurones from older embryos was greatly increased, reaching a maximum of over 80% for 8-day embryonic motoneurones. In contrast, the number of motoneurones that had regenerated neurites when co-cultured with dorsal root ganglion cells for 24 h decreased with the motoneurone age at plating, from 51% at 5 days to less than 10% for 7- and 8-day motoneurones. The survival-enhancing effects were probably mediated by cell contact between the motoneurones and processes of the dorsal root ganglion cells: conditioned media from high-density cultures of dorsal root ganglion cells could not be shown to significantly enhance the survival of motoneurones above that of control levels. The possibility that the ganglion cells exert this survival enhancing effect by depolarizing the motoneurones was examined by exposing 8-day sorted motoneurones to 47 mM potassium; this did not effect the survival of the motoneurones relative to control levels. The stage dependency of the survival of motoneurones on different neurotrophic factors and the dorsal root ganglion cell is discussed. PMID- 3032373 TI - Morphine- and ketocyclazocine-induced analgesia in the developing rat: differences due to type of noxious stimulus and body topography. AB - Patterns of morphine- and ketocyclazocine-induced analgesia in limb withdrawal and tail-flick tests of thermal and mechanical nociception were examined in the preweanling rat. In the forepaw test, morphine was more effective than ketocyclazocine with both thermal and mechanical stimuli. Both drugs first induced analgesia between 3 and 5 days of age. In the tail-flick test, ketocyclazocine-induced analgesia preceded morphine's effects against both thermal and mechanical stimuli by several days. Ketocyclazocine produced robust analgesia between 7 and 10 days of age, while the effects of morphine did not peak until day 14. In the hindpaw, morphine was more effective than ketocyclazocine against a higher intensity mechanical stimulus, while ketocyclazocine was more effective against a lower intensity mechanical stimulus. Morphine-induced analgesia was reversed by lower doses of naloxone than was ketocyclazocine-induced analgesia, regardless of body part tested, against all noxious stimuli. These findings demonstrate differences in morphine- and ketocyclazocine-induced analgesia that are dependent upon age, body topography, stimulus type and intensity and imply different physiologic roles of mu- and chi opioid receptors in analgesia. PMID- 3032374 TI - Neonatal 6-hydroxydopamine lesion of spinal noradrenergic terminals: nociception, clonidine analgesia and spinal alpha two adrenoceptors. AB - Newborn rats received intraspinal injections of 6-hydroxydopamine to enduringly deplete spinal norepinephrine (NE). When tested in adulthood for pain sensitivity with a hot water-tail immersion procedure, this neonatal spinal NE lesion lowered tail flick latencies of females but not males. It was postulated that this sexually dimorphic sparing or recovery of function reflected the development of denervation supersensitivity for males but not females. Contrary to expectation from such an hypothesis, females, not males, showed exaggerated sensitivity to the analgesic effects of a test dose of clonidine. Furthermore, neither males nor females showed an increased number of spinal cord binding sites for (3H)para amino-clonidine [(3H)PAC]. These receptor binding data failed to indicate proliferation of the spinal alpha two adrenoceptor in either sex. That the lesioning of spinal NE terminals did not reduce (3H)PAC binding sites suggests that the spinal alpha two adrenoceptor does not reside exclusively on NE terminals. This is consistent with current conclusions concerning the alpha two adrenoceptor in the cerebral cortex. PMID- 3032376 TI - [Note on certain hereditary neuropathies involving hypersensitivity to pressure: sausage-like neuropathies]. PMID- 3032375 TI - Tryptamine receptors: fact, myth or misunderstanding? AB - Tryptamine is an endogenous brain amine which is implicated in neural regulation and proposed to play a significant role in the aetiology of some neuropsychiatric illnesses. Recent reports indicate the possible existence of specific tryptamine binding sites. It has been postulated that these binding sites may be functional tryptamine receptors in the central nervous system. The status of current developments in this area is critically reviewed. Current problems are outlined and discussed in terms of the specificity of the [3H]-tryptamine binding site and its functional assessment with experiments involving both drug treatment and electrolytic and neurotoxin-induced brain lesions. Current data indicate that the [3H]-tryptamine binding site is selective and not attributable to residual monoamine oxidase binding. PMID- 3032377 TI - Diseases of the peripheral nerves. AB - Peripheral nerve disorders involving the bovine thoracic and pelvic limbs are presented. Anatomic considerations, etiologic mechanisms, clinical signs, diagnosis, and prognosis are detailed for each specific impairment. Treatment, although generally supportive, is also discussed. PMID- 3032378 TI - Prevention of parathyroid hormone-dependent nephrocalcinosis by chronic administration of the organic phosphorothioate WR-2721. AB - Pharmacological inhibition of parathyroid hormone (PTH) secretion has been the object of several experimental and clinical trials in the past. It is only recently that a drug, WR-2721 [S-,2-(3-aminopropylamino)-,ethylphosphorothioic acid], has been shown to effectively inhibit PTH secretion in euparathyroid subjects and in parathyroid cancer patients within a few hours after its administration. In the present study we tested its long term efficacy as a PTH inhibitor in a model of nephrocalcinosis resulting from secondary hyperparathyroidism. Intact and thyroparathyroidectomized (TPTX) rats were pair fed on a low Ca (0.2%)-high phosphorus (1.6%) diet (nephrocalcinotic diet) or on a normal Ca (1.1%)-normal phosphorus (0.8%) diet for 7 days. Simultaneously, either 0.2 mmol/kg of WR-2721 or its solvent was injected subcutaneously twice daily. The intact animals on the nephrocalcinotic diet had an increased urinary cyclic AMP excretion and important renal Ca accumulation. This nephrocalcinosis was markedly reduced by WR-2721 treatment. The kidney Ca content of the WR-2721 treated rats was 58 +/- 6% lower than that of the nontreated animals. The low Ca high phosphorus diet did not cause nephrocalcinosis in the TPTX rats. WR-2721 failed to reduce the nephrocalcinosis induced by 1,25 dihydroxyvitamin D3 intoxication in TPTX rats fed the normal Ca-normal phosphorus diet. In conclusion, the present study suggests that chronic treatment with WR-2721, a potent inhibitor of PTH secretion, may be effective for preventing the deleterious consequences of hyperparathyroidism, such as nephrocalcinosis. PMID- 3032379 TI - Occurrence of nitrogenous species in precipitated B-type carbonated hydroxyapatites. AB - B-type carbonated hydroxyapatites, prepared in aqueous media free of alkali ions, fix ammonium ions present in the reaction medium. A small portion of the carbonate ions introduced into the apatite structure enter by the substitution mechanism (CO3(2-), NH4+)----(PO4(3-), Ca2+). With these results for the structural incorporation of ammonium ions, differences in lattice parameters observed among specimens with the same degree of carbonation were attributed to some substitution of NH4+ for Ca2+. The fixed ammonium ions were shown to be the source of the cyanamide and cyanate ions that develop on heating. Above 500 degrees C these apatites lost both the carbonate and the cyanate and cyanamide ions. PMID- 3032381 TI - The effects of gestational age and intrafetal ACTH administration on the concentration of progesterone in the fetal membranes, endometrium, and myometrium of pregnant sheep. AB - To examine the hypothesis that progesterone withdrawal from intrauterine tissues is a prerequisite to spontaneous labour or labour induced by administering ACTH to the ovine fetus, we measured the concentration of progesterone in amnion, chorion, endometrium, and myometrium of sheep at different stages of pregnancy and during ACTH-induced labour. There was no significant change in the concentration of progesterone nor in the progesterone:estradiol ratio in amnion or chorion in association with either spontaneous or ACTH-induced labour. The concentration of progesterone in endometrium rose significantly between days 50 60 and days 130-135 of gestation and decreased at term. There was also a fall in the progesterone:estradiol ratio in endometrium between days 130-135 and term. Neither the progesterone concentration not the progesterone:estradiol ratio changed in endometrium during ACTH-induced labour. In the myometrium the concentration of progesterone rose significantly between days 50-60 and day 100 of pregnancy and decreased between day 100 and days 130-135, with a further decline towards term. After intrafetal ACTH there was no change in the concentration of progesterone in the myometrium, although there was a fall in the progesterone:estradiol ratio. We conclude that labour occurring spontaneously at term is associated with a decrease in the progesterone concentration of maternal intrauterine tissues, the myometrium and endometrium. In contrast, there is no decline in the progesterone concentrations of the fetal membranes, the amnion and chorion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3032380 TI - Tissue-specific concentration changes of estrone and estradiol during spontaneous and ACTH-induced parturition in sheep. AB - Changes in estrogen production are considered important in the sequence of events leading to parturition. We sought tissue-specific changes in the concentration of unconjugated estrone (E1) and estradiol (E2) in intrauterine fetal (amnion, chorion) and maternal (endometrium, myometrium) tissues during normal pregnancy, labour, and ACTH-induced labour in sheep. The mean concentrations of E1 and E2 in the fetal membranes were higher than in endometrium and myometrium. In amnion there were no consistent changes in estrone concentrations with gestation, although estradiol concentrations increased between day 130 and term. In the endometrium there were increases in both estrone and estradiol between day 100 and term, whereas in the myometrium increases in the concentrations of E1 and E2 occurred between days 130-135 and term. Animals showing a labourlike pattern of uterine contractions after intrafetal ACTH administration did not show significant differences in estrone or estradiol concentrations in amnion, chorion, or endometrium compared with saline-infused controls. However, there was a progressive increase in the concentration of estrone and estradiol in the myometrium during ACTH-induced labour. We conclude that changes in the concentrations of estrone and estradiol in intrauterine tissues vary between the tissues studied and the two estrogens. In general, estrogen concentrations increased towards term, but this trend was more marked in the maternal than fetal tissues. The changes in estrone concentrations in myometrium, but not in the other tissues, were replicated during ACTH-induced labour. Our results would be compatible with the suggestion that tissue-specific changes in estrogen concentrations may contribute to the local intrauterine steroid milieu during pregnancy and at term. PMID- 3032382 TI - Effects of a specific bradycardic agent (AQ-A39) and verapamil on beta-adrenergic responses in isolated guinea pig atria. AB - AQ-A39 (5,6-dimethoxy-2-[3-[(3,4-dimethoxy)phenylethyl)methylamino]propyl)- phthalimidine), a specific bradycardic agent, and verapamil, a calcium channel blocker, were studied for their ability to alter rate and force of contraction in the presence and absence of isoproterenol, a beta-adrenergic stimulant, using isolated guinea pig atria. Both compounds (10(-7)-10(-4) M) produced dose-related decreases in frequency of spontaneously beating right atria. Verapamil decreased, while AQ-A39 increased, the force of contraction of electrically stimulated (1.0 Hz) left atria. At equal negative chronotropic concentrations, AQ-A39 was more effective than verapamil in reducing the maximum isoproterenol-induced tachycardia. Verapamil, but not AQ-A39, antagonized positive inotropic responses to isoproterenol. Therefore, AQ-A39 differed from verapamil in that (i) AQ-A39 was a more selective bradycardic agent in both beta-adrenergically stimulated and nonstimulated preparations and (ii) AQ-A39 was more effective in reducing isoproterenol-elevated heart rate compared with basal heart rate. This profile of activities suggests that AQ-A39 will be beneficial in cardiac pathologies where sympathetic nervous system activity is elevated and a lowering of heart rate without a reduction in cardiac contractility is desired. PMID- 3032383 TI - Prolonged isobaric relaxation time in small mesenteric arteries of the spontaneously hypertensive rat. AB - Prolonged isometric relaxation in hypertensive aortic and caudal arterial smooth muscle has been demonstrated; however, isobaric relaxation in resistance arteries is more pertinent to studies in hypertension. A comparative study of mesenteric arterial isobaric relaxation times was made using spontaneously hypertensive rats (SHR), normotensive Wistar-Kyoto rats (WKY), and MK-421 treated SHR (treatment commenced at 8 weeks of age and was maintained until sacrifice). Relaxation rates of vessels constricting against a range of pressures and achieving different degrees of narrowing or changes in circumference were analyzed. Comparisons were made between SHR, WKY, and MK-421 treated SHR arteries that had constricted from the same initial circumference and against the same magnitude of pressure. The SHR mesenteric arteries relaxed at a slower rate than did the WKY vessels. The normotensive MK-421 treated SHR showed the same prolonged relaxation rate as did the untreated SHR preparations. Thus the slower rate of relaxation in SHR arteries does not appear to be a consequence of the hypertension. Such prolonged time for narrowing would function to increase the average peripheral resistance and thus may contribute to the initiation and maintenance of increased blood pressure. PMID- 3032384 TI - Effect of cold exposure on energy metabolism in the young pig. AB - Twenty-four piglets were weaned at 3 weeks of age and received 615 kJ metabolizable energy/(kg body weight X day). The temperature was reduced from 29 to 25 degrees C by 4 weeks of age. Six pigs were exposed to cold by reducing the temperature to 10 degrees C by 5 weeks of age while the other six were exposed to 23 degrees C. These two temperatures were maintained for 3 weeks. Each week the three pigs closest to the mean weight of each group were used for measurements of heat production for 23 h by open-circuit calorimetry and glucose turnover by continuous infusion of [6-3H]- and [U-14C]-glucose. The animals were sacrificed at 8 weeks of age, a sample of their intercostal muscle was used for in vitro measurement of muscle respiration, and the carcass was analyzed. There was no change in heat production, glucose turnover, and rectal temperature during the 3 weeks of cold exposure, so the data were pooled. Cold exposure increased heat production by 50%, forced mobilization of fat reserves (1000 g over the 3 weeks), and increased glucose replacement rate by 20%. The decline in rectal temperature to 37.6 from 38.8 degrees C could be regarded as a strategy to reduce energy needed to regulate body temperature. In the cold, heat production equalled metabolizable energy intake, but protein deposition was maintained at the same level as that in the pigs at the thermoneutral temperature, using the energy derived from the mobilization of body fat. The increase in Na+-K+ ATPase dependent respiration accounted for 70% of the increase in O2 consumption of muscle from cold-exposed pigs and thus is potentially an important component of cold-induced thermogenesis in the pig. PMID- 3032385 TI - Characterization of two abundantly expressed constitutive genes of Neurospora crassa. AB - Two abundantly expressed, constitutive genes of Neurospora crassa were isolated during differential screening of Neurospora genomic libraries. The coding regions of these two genes, designated RLF1 and RLF3, were identified by hybridization of the cloned DNA sequences with cDNA probes made from polyadenylated RNA. The RLF3 gene was carried on a 15-kilobase Neurospora BamHI DNA fragment present in a lambda 1059 recombinant; a 2-kilobase restriction fragment that contains RLF3 was subcloned into plasmid pBR322 prior to further characterization. Southern blot analysis revealed that both RLF1 and RLF3 are single copy genes. Northern blot analysis and S1 nuclease mapping demonstrated that RLF1 is transcribed to yield a 1.6-kilobase RNA, whereas RLF3 appears to give rise to two distinct sized RNA species of 1.0 and 1.6 kilobases. RNA dot blot analysis provided conclusive evidence that both of these genes are constitutively expressed. These constitutive genes will be valuable to provide a detailed comparison with the 5' flanking regions of regulated genes. PMID- 3032386 TI - Oligonucleotide probes for the detection of TEM-1 and TEM-2 beta-lactamase genes and their transposons. AB - We describe the use of molecular probes to detect the TEM-type beta-lactamase genes. As a general probe, we prepared a 656 base pair restriction fragment, entirely within the TEM structural gene. This probe was specific for the TEM family, hybridizing only with TEM-1 and TEM-2. The TEM-1 and TEM-2 beta lactamases differ by only one amino acid. We synthesized two oligonucleotides whose central bases correspond to this difference. The use of these oligonucleotides enables us to discriminate between TEM-1 and TEM-2 genes. Using oligonucleotides homologous to parts of Tn3, we also monitored the presence of TnA-like transposons in bacteria harboring different beta-lactamase genes. Only the TEM-1 and TEM-2 genes were found to be on transposons with terminal sequences identical to those of Tn3. All hybridization experiments were performed with both dot-blot and colony-hybridization techniques, and the suitability of these two methods for epidemiological studies is compared. PMID- 3032387 TI - Genome homology and divergence in the SP beta-related bacteriophages of Bacillus subtilis. AB - Chromosomal organization in related temperate Bacillus subtilis bacteriophages SP beta, phi 3T, rho 11, Z, and E was compared. DNA-DNA hybridization studies done in conjunction with available restriction fragment maps of SP beta, phi 3T, and rho 11 demonstrated that DNA homology between these three phages extended over most of their respective genomes, although each contained unique chromosomal segments, phi 3T, rho 11, Z, and E, but not SP beta, possessed apparently homologous structural genes (thyP) for thymidylate synthetase. DNA from all thyP containing phages transformed thymine auxotrophs of B. subtilis SP beta lysogens to prototrophy. This transformation commonly involved incorporation of the thyP gene into SP beta prophage within a region corresponding to the middle of the viral chromosome. Chimeric plasmids containing the thyP gene from phi 3T or cloned fragments of SP beta DNA were used in DNA-DNA hybridization studies to locate the thymidylate synthetase gene near the center of the phi 3T chromosome, and to demonstrate that the organization of this region resembled the analogous portion of the SP beta genome. Profiles of virion structural proteins from the five phages were also very similar, further suggesting functional homology between these viruses. However, despite these evidences of relatedness, populations of fragments generated by digesting SP beta, phi 3T, rho 11, Z, and E DNA with restriction enzymes were quite dissimilar. PMID- 3032388 TI - A necrotizing pneumonia in lambs caused by pseudorabies virus (Aujesky's disease virus). AB - An outbreak of pseudorabies occurred in sheep housed with swine in the same building. Although the sheep and swine were not in physical contact, the lambs and ewes were exposed to air from the sows' section. Three dead lambs were submitted to the Iowa State University Veterinary Diagnostic Laboratory for necropsy. Grossly there were pulmonary congestion and multifocal pulmonary hemorrhages. Microscopic lesions were severe acute multifocal necrotizing bronchopneumonia with necrotizing vasculitis and intranuclear inclusion bodies within the neurons of the parabronchial ganglia. Bacterial cultures were negative for pathogenic agents; pseudorabies virus was isolated from ovine brain tissue. Viral antigen was demonstrated in the neurons of the parabronchial ganglia by immunoperoxidase staining. Electron microscopy revealed nucleocapsids in the parabronchial ganglionic neurons which contained basophilic intranuclear inclusion bodies. Viral DNA prepared from the ovine pseudorabies virus isolate was found by restriction endonuclease analysis to be related to the Indiana Funkhauser strain of pseudorabies virus. PMID- 3032390 TI - The kinetics-based enzyme-linked immunosorbent assay for coronavirus antibodies in cats: calibration to the indirect immunofluorescence assay and computerized standardization of results through normalization to control values. AB - The computer-assisted, kinetics-based enzyme-linked immunosorbent assay for coronavirus antibodies in cats was calibrated to the conventional indirect immunofluorescence assay by linear regression analysis and computerized interpolation (generation of "immunofluorescence assay-equivalent" titers). Procedures were developed for normalization and standardization of kinetics-based enzyme-linked immunosorbent assay results through incorporation of five different control sera of predetermined ("expected") titer in daily runs. When used with such sera and with computer assistance, the kinetics-based enzyme-linked immunosorbent assay minimized both within-run and between-run variability while allowing also for efficient data reduction and statistical analysis and reporting of results. PMID- 3032389 TI - Vaccination of calves with a diaminopimelic acid mutant of Salmonella typhimurium. AB - The purpose of this study was to evaluate the safety and efficacy of a diaminopimelic acid mutant of Salmonella typhimurium as a vaccine for calves. Transposon techniques were used to create a stable nonreverting diaminopimelic acid mutant of a virulent S. typhimurium strain. Calves were vaccinated at weekly intervals with the diaminopimelic acid mutant given as an oral dose of 10(10) organisms, followed by two subcutaneous doses of 10(9) organisms. The calves tolerated vaccination well and the vaccine strain was eliminated from the feces within four days. Of five vaccinated calves, three survived challenge with 5 X 10(9) organisms of the parent strain whereas all five unvaccinated calves that were challenged died. The surviving calves eliminated the challenge organism from the feces within three weeks. PMID- 3032391 TI - Suppression of cortisol responses to exogenous adrenocorticotrophic hormone, and the occurrence of side effects attributable to glucocorticoid excess, in cats during therapy with megestrol acetate and prednisolone. AB - The major purpose of this investigation was to determine the effect of prednisolone and megestrol acetate in cats on the adrenal cortisol response to exogenous adrenocorticotrophic hormone during drug administration at dose rates employed for management of some inflammatory feline dermatoses. Prednisolone (at least 2 mg/kg/day) and megestrol acetate (5 mg/cat/day) were each administered orally to seven cats from days 1 to 16. Three additional cats received no therapy. Basal and stimulated cortisol concentrations, food and water intake, hematology, blood biochemistry, urinalyses, and hepatic and cutaneous histology were studied in all cats before, during, and two weeks following the end of treatment. Cats given prednisolone or megestrol acetate had significant suppression of stimulated cortisol levels on day 8. This change was more marked on day 15, when the suppression in cats given megestrol acetate was also significantly more severe than in those receiving prednisolone. Recovery of adrenal reserve was considered present on day 30 in six of seven cats given prednisolone, but in only three of seven receiving megestrol acetate. Eosinopenia, glycosuria and hepatocyte swelling from glycogen deposition were occasionally recorded in treated cats of both groups, providing additional circumstantial evidence for glucocorticoid activity of megestrol acetate in cats. It is advised that abrupt withdrawal of prednisolone or megestrol acetate therapy be avoided in this species to reduce the chance of precipitating clinical signs of hypoadrenocorticism, even after treatment for as little as one week. PMID- 3032393 TI - The inpatient treatment of the borderline personality disorder: a critical review and discussion of aftercare implications. AB - The literature on inpatient treatment of adult borderline personality disorder is reviewed. The controversy regarding the use or avoidance of the regression which can accompany hospitalization is discussed. It is suggested that this literature is not specific enough. There is insufficient attention to definition of terms, clinical heterogeneity and the longitudinal course of the illness. As well, the ahistoric approach to the hospitalization of these patients serves to reinforce characteristic ego deficits. Recommendations regarding the use of long or short term length of hospital stay are not substantiated by formal research. It is suggested that greater emphasis be placed on aftercare issues. The stabilization of the patient, outpatient therapist, and aftercare system are described as important goals of inhospital care. PMID- 3032392 TI - Stability of bovine leukemia virus antigens. AB - Stability of bovine leukemia virus glycoprotein 51 and nonglycosylated protein 24 antigens was examined under various conditions. The glycoprotein antigen was unstable at 56 degrees C and 37 degrees C and at acidic and basic pH. The specificity of the glycoprotein 51 antigen was reduced to half by the first treatment with ethyl ether, but was not decreased further by repeated treatments. This antigen was not inactivated with triton X-100 and was resistant to lyophilization as well as to freezing and thawing. The protein 24 antigen was generally very stable under all conditions tested. PMID- 3032394 TI - Long-term observations of the patterns of failure in patients with unresectable non-oat cell carcinoma of the lung treated with definitive radiotherapy. Report by the Radiation Therapy Oncology Group. AB - This report details the patterns of tumor recurrence in two prospective randomized studies involving 551 patients with histologically proven unresectable or medically inoperable non-oat cell carcinoma of the lung treated with definitive radiotherapy. Patients were treated according to two protocols, depending on the stage of the tumor: (1) Patients with T1, 2, 3-NO, 1, 2 tumors were randomized to four different regimens: 4000 cGy split course (2000 cGy in five fractions, per 1 week, 2 weeks rest and additional 2000 cGy in five fractions, per 1 week) or 4000, 5000, or 6000 cGy continuous courses, five fractions per week. (2) Patients with T4, any N or N3, any T stage tumors were randomized to be treated with 3000 cGy tumor dose (TD), ten fractions in 2 weeks, 4000 cGy split course (described above), or 4000 cGy continuous course. In the patients with less advanced tumors (Study 1) the intrathoracic failure rate within the irradiated volume was 48% with 4000 cGy continuous, 38% with 4000 cGy split course or 5000 cGy continuous, and 27% for patients receiving 6000 cGy continuous course. The failure rate in the nonirradiated lung ranged from 25% to 30% in the various groups. Patients with adenocarcinoma or large cell undifferentiated carcinoma had better intrathoracic tumor control (35%) than those with squamous cell carcinoma (20%). The incidence of distant metastases was 75% to 80% in all histologic groups. Distant metastases appeared sooner after therapy in the patients with adenocarcinoma or large cell undifferentiated carcinoma. The initial failure rate in the brain was 7% in patients with squamous cell carcinoma, 19% with adenocarcinoma, and 13% in patients with large cell carcinoma. The ultimate incidence of brain metastases was 16% in squamous cell carcinoma, and 30% for adenocarcinoma or large cell undifferentiated carcinoma. Higher doses of irradiation will be necessary in order to improve the intrathoracic tumor control. Clinical trials by the Radiation Therapy Oncology Group, some of them involving multiple daily fractionation, are in progress. Furthermore, because of the high incidence of distant metastases, effective systemic cytotoxic agents are critically needed to improve survival of lung cancer patients. The high frequency of brain metastases suggests that, as in small cell carcinoma of the lung, elective irradiation of the brain may be necessary, if not to improve survival to enhance the quality of life of patients with adenocarcinoma and large cell carcinoma. PMID- 3032395 TI - Angioinvasion in breast carcinoma. An immunohistochemical study of factor VIII related antigen. AB - Histologic sections from 63 patients with infiltrating duct carcinoma of the breast were selected for study by immunohistochemical staining with antibody against human Factor VIII-related antigen. Of these 63, 30 had no lymph node metastases at the time of surgery, while 33 had axillary lymph node metastases. A positive correlation exists between the presence of vascular invasion and lymph node metastases. Sixty-nine percent of patients with lymph node metastases had vascular invasion while only 26% of patients without lymph node metastases showed evidence of invasion of blood vessels. The finding of vascular invasion by tumor in patients without axillary lymph node metastases at the time of mastectomy may explain the occurrence of disseminated disease years after treatment. PMID- 3032396 TI - Flow cytometric analysis of DNA in bone and soft-tissue tumors using nuclear suspensions. AB - Ninety-four bone and soft tissue tumors were analyzed for their DNA content using flow cytometry (FCM). A simple, rapid method for preparing isolated nuclear suspensions was used. Tissues, minced in a hypotonic solution containing detergent and propidium iodide as a fluorescent probe for DNA, provided in most instances high nuclear yields from only 0.02 to 0.03 g of solid tumor. Whereas all nonneoplastic samples had a diploid DNA content, various degrees of abnormal DNA distributions were detected in 90% of the neoplastic samples and were present in benign as well as malignant tumors. Our findings demonstrate that FCM DNA analysis is practical in most musculoskeletal tumors and support the observations of others that abnormal DNA content may serve as a general neoplastic marker in these tumors. PMID- 3032397 TI - Anaplastic sarcoma arising in a mature metachronous bilateral Wilms' tumor after irradiation and chemotherapy. Spontaneous versus induced malignant change. AB - A 1-year-old male infant developed a classic Wilms' tumor of the left kidney. Treatment consisted of a left nephrectomy, chemotherapy, and irradiation to the left flank and associated abdomen. Two years later, a mass in the right kidney was discovered; open renal biopsy demonstrated a mature Wilms' tumor consisting entirely of rhabdomyomatous elements in the biopsy specimen. The patient was given a second course of chemotherapy and 2000 rad to the right flank. Over the next 8 years, the mass continued to grow without evidence of metastatic spread. Renal function deteriorated secondary to compression of the surrounding normal renal parenchyma by the enlarging tumor; creatinine clearance from the solitary kidney decreased from 120 ml/min to 40 ml/min during the 12 months prior to removal of the lesion. Via a nephron-sparing procedure, the 3400 g tumor measuring 19 cm X 16 cm X 9 cm was enucleated from the right kidney without compromise to the remaining normal tissue. Pathologic examination of the surgical specimen revealed a mature Wilms' tumor with a malignant anaplastic sarcoma arising in the central portion. Postoperatively, the patient received a third course of chemotherapy with no irradiation to the tumor bed. Currently, he is disease-free with normal renal function more than 20 years after diagnosis of the metachronous bilateral Wilms' tumor. This is the first reported case of an anaplastic sarcoma arising within a Wilms' tumor; this individual also is the longest surviving patient with metachronous Wilms' tumor. The various possibilities regarding the development of the anaplastic sarcoma within the Wilms' tumor of the right kidney are discussed, including the possible role of chemotherapy and irradiation in the development of a second malignancy. PMID- 3032399 TI - The characteristics of embryonal carcinoma cells in teratocarcinomas. AB - The orchiectomy specimens and the respective lymphadenectomies of 33 teratocarcinomas (teratoma and embryonal carcinoma) and 30 embryonal carcinomas were identified in a series of 457 consecutive germ cell tumors of the testis. Although teratocarcinomas were larger tumors the retroperitoneal lymphadenectomies revealed metastases in only 10 of 33 (30%) teratocarcinomas as compared to 19 of 30 (63%) embryonal carcinomas. Even after subtracting the teratoma component and stratifying for size of the embryonal carcinoma component, the teratocarcinomas were still less likely to metastasize than comparably sized pure embryonal carcinomas. Statistical significance was found between the differences in size and the differences in the rates of metastases, before and after stratifying for size of the embryonal carcinoma component of the teratocarcinomas. Further, all embryonal carcinomas metastasized as embryonal carcinoma while only 5 teratocarcinomas metastasized as embryonal carcinoma. This study supports experimental evidence that the embryonal carcinoma cells in teratocarcinomas are not necessarily identical to embryonal carcinoma cells in embryonal carcinomas. PMID- 3032398 TI - Antigenic phenotype of the lymphocytic component of medullary carcinoma of the breast. AB - Medullary carcinoma of the breast, which is usually associated with a dense lymphocytic infiltrate, carries a better prognosis than do most other histologic subtypes of breast carcinoma. We studied cryostat-cut fresh frozen sections from 12 patients with medullary carcinoma and, as controls, nine patients with infiltrating ductal carcinoma in order to determine and compare the antigenic phenotype of the lymphocytic components of these tumors. We used a large panel of monoclonal antibodies and polyclonal antisera for T-cells (Leu-1, Leu-2a, Leu-3a, Leu-9, T-3, T-6, T-10, T-11, and TQ-1), pre-B and B-cells (BA-1, B-1, B-2, B-4, and J5), NK cells (Leu-7 and Leu-11b), and cell activation associated antigens (T 9, HLA-Dr, and Tac). The most commonly encountered antigens on the lymphocytic components of both medullary carcinoma and infiltrating ductal carcinoma were: T 3, T-11, Leu-1, Leu-2a, Leu-3a, and Leu-9. There was little staining for NK-, pre B-, or B-cell associated antigens in either type of carcinoma. However, the lymphocytes in the control cases tended to express HLA-Dr and T-10 more often than did the lymphocytes in the cases of medullary breast carcinoma. Our data indicate that: the antigenic phenotypes of the lymphocytic infiltrates of medullary carcinoma and those of infiltrating ductal carcinoma of the breast are essentially similar; and the lymphocytes in these carcinomas are composed predominantly of peripheral T-lymphocytes. We therefore conclude that the favorable biologic behavior of medullary carcinoma of the breast cannot readily be explained by the immunophenotype of its lymphocytic component. PMID- 3032400 TI - Use of carcinoembryonic antigen in small cell lung cancer. Prognostic value and relation to the clinical course 1. AB - Carcinoembryonic antigen (CEA) was measured in 147 patients at diagnosis of small cell lung cancer; 17% of patients with limited disease and 51% with extensive disease had an abnormal CEA level (greater than 10 ng/ml). The median level was higher in extensive than in limited disease (11 ng/ml and 5.8 ng/ml, respectively; P less than 0.001). Multivariate analysis showed CEA level greater than or equal to 50 ng/ml to be an adverse prognostic factor (P = 0.02); median survival at this level was shorter than at less than 50 ng/ml (7 and 46 weeks, respectively; P = 0.002). No consistent directional changes of follow-up CEA values were observed in patients with initially normal CEA levels, but normalization of levels occurred in complete responders. We recommend that CEA be measured in this disease at diagnosis as an additional prognostic factor and that patients with abnormal initial CEA levels have follow-up measurements to aid in evaluating response. PMID- 3032401 TI - Metastatic islet cell tumor with ACTH, gastrin, and glucagon secretion. Clinical and pathologic studies with multiple therapies. AB - A patient with metastatic islet cell carcinoma demonstrated multiple clinical syndromes simultaneously with secretion of ACTH, gastrin, glucagon, and serotonin. Hepatic arterial embolization resulted in an initial decrease in all secretory products, which was sustained for glucagon and serotonin. Recrudescence of the Cushings and Zollinger-Ellison syndrome was managed by surgical extirpation of the primary tumor and regional metastases as well as bilateral adrenalectomy. Electron microscopy and immunocytochemistry of the primary tumor and the metastatic lesions revealed the presence of multiple types of granules within single cells and, different patterns of secretory profiles in different tumor sites. PMID- 3032402 TI - Topographic study of cervical condyloma and intraepithelial neoplasia. AB - In this study of 101 cervical conization specimens, the location and the size of condyloma and intraepithelial neoplasia (dysplasia and carcinoma in situ) were mapped using the last endocervical gland as the marker for the original squamocolumnar junction. Condylomatous changes were identified in 85% of cervices affected by the intraepithelial neoplasia, and were in direct contact with 68% of intraepithelial neoplasms. The proximal location of intraepithelial neoplasia in relation to the condyloma can be explained by the occurrence of neoplasia just proximal to the condyloma and subsequent expansion of neoplasia towards the cervical canal. This study adds topographic evidence linking cervical condyloma to the development of intraepithelial neoplasia. PMID- 3032403 TI - Prognostic factors in patients with hepatocellular carcinoma. Attempts for the selection of patients with prolonged survival. AB - A prognostic study based on 127 untreated patients with hepatocellular carcinoma was undertaken to evaluate their survival time and to find clinical and biologic criteria which allow the selection of patients with a survival time longer than 60 days who could enter a therapeutic trial. Twenty-eight clinical and biologic variables were assessed using univariate and multivariate semiparametric regression (Cox's) models. Ten variables were isolated by univariate analysis. Multivariate analysis found a negative relationship between a survival time longer than 60 days and five of these variables; these variables were in decreasing order: encephalopathy, alcohol consumption, aspartate amino transferase (AST), blood urea nitrogen, and total bilirubin. Prevalence, positive, and negative predictive values of encephalopathy were 20%, 27.5%, and 97% respectively. When three other criteria: ASAT greater than four times the upper limit of the normal (N), blood urea nitrogen greater than N, and total bilirubin greater than 2N were added, their prevalence, positive, and negative predictive values were 72%, 89.7%, and 57.1% respectively. These results suggest that in countries where incidence of hepatocellular carcinoma is low and recruitment of patients difficult, absence of encephalopathy must be the only criterion for selection of patients with hepatocellular carcinoma in therapeutic trials; whereas, in countries with a high incidence of hepatocellular carcinoma the other criteria may be added. PMID- 3032404 TI - Karyotypic characterization of an established human hepatoma cell line HA22T/VGH. AB - The karyotype of an established human hepatoma cell line HA22T/VGH was characterized by G-banding. A majority of the 200 cells counted had around 70 chromosomes at passage 24, and 60 at passage 338. Of the 50 cells karyotyped from each of passage 24 and passages 338-339, chromosomes #13 and #18 were absent. The presence of the Y chromosome was reduced dramatically from a mean value of 1.12/cell at passage 24 to 0.12/cell at passages 338-339. In general, most of the chromosomes--particularly chromosomes #5, #7, #9, #15, and #21--tended to be less represented in the course of propagation in vitro. The presence of multiple copies of a normal chromosome in a single cell was quite common for chromosomes #5 and #7 at both early and late passages. Numerous structural rearrangements of the chromosomes were observed. PMID- 3032405 TI - Abnormal chromosomes including small metacentrics in 14 ovarian cancers. AB - In direct preparations of 14 ovarian cancers including 11 primary tumors, chromosomes #1 (12 tumors), #3 (12 tumors, including 3q- chromosomes in five), #6 [eight tumors, including six with a 6q- and two with an i(6p)], #11 (11p + in seven tumors), and #14 (14q+ in at least seven tumors) were most frequently involved in structural aberrations. Also, abnormal small metacentrics were seen in 11 tumors. In ten of these the chromosome appeared to be an i(4p) or i(5p) and in one of these, a mixed Mullerian tumor, there was also an i(12p); the latter anomaly was also present (in duplicate) in a dysgerminoma. PMID- 3032406 TI - Metabolic activation of diethylstilbestrol by stimulated human leukocytes. AB - Previous studies have implicated both peroxidases and leukocytes in the metabolic activation of the human carcinogen diethylstilbestrol (DES). Here we demonstrate that DES is converted during the oxidative burst of human leukocytes to reactive protein binding species. Although luminol-dependent chemiluminescence indicated that peroxidase-dependent metabolism was occurring, the protein binding was not inhibitable by azide. This suggested that either peroxidase-mediated metabolism was not responsible for the formation of the reactive protein binding species or that this binding was occurring in a cellular compartment inaccessible to azide. The addition of catalase alone and in combination with superoxide dismutase (SOD) did, however, result in significant inhibition of binding. Hypochlorous acid was also shown to be capable of directly converting DES to protein binding species. These results indicate that a product of the oxidative burst, most likely a highly oxidizing species derived from H2O2, is capable of converting DES to a potentially carcinogenic binding species. PMID- 3032407 TI - Clonal evolution of t(14;18) follicular lymphomas demonstrated by immunoglobulin genes and the 18q21 major breakpoint region. AB - A 2.8-kilobase major breakpoint region on chromosome segment 18q21 is the site of most t(14;18) translocations typical of human follicular lymphomas. Breaks are focused at the 5' end of joining (JH) regions of immunoglobulin (Ig) on chromosome 14, indicating that the translocation occurs at a pre-B-cell stage during attempted heavy (H) chain joining. A new gene from 18q21 (Bcl-2) is placed in the H chain locus creating a unique, translocation-specific JH;18q21 rearrangement that presumably represents a transformation event. In addition, normal Ig gene joining occurs in a H before light (L) chain and K before lambda cascade, creating ordered clonal markers. These serial markers were examined to determine if variations in Ig gene patterns during the natural history of lymphomas represent the emergence of truly separate neoplasms or heterogeneity of a single neoplasm. We examined 45 serial biopsies from 16 B follicular lymphoma patients; six cases showed variation in Ig gene patterns over time. Seven individuals had a detectable JH;18q21 rearrangement present, and it remained unchanged over 5-10 years. Further rearrangements of H chain genes occurred on the normal chromosome 14 within evolving subclones of the original tumor. Lambda L chains also underwent additional rearrangements in two instances, while K gene patterns remained unchanged. All variations in the normal H and L chain genes were 2 degrees rearrangements occurring at a mature B-cell stage following the initial successful rearrangement of a H and L chain. In contrast the t(14;18) breakpoint was conserved in each individual, indicating that evolving neoplastic subpopulations arose from a common clonal progenitor cell. PMID- 3032408 TI - Lysis by activated lymphocytes of melanoma and small cell lung cancer cells surviving in vitro treatment with mafosfamide. AB - Six short term-cultured melanoma cell lines and one small cell lung cancer cell line were treated in vitro with the alkylating agent mafosfamide. The sensitivity of the surviving cells to in vitro lysis by recombinant interleukin 2-activated autologous and allogeneic lymphocytes was then investigated. In no case did chemo surviving tumor cells appear less sensitive to lymphocyte-mediated lysis than untreated counterparts. In three of seven cases (two of which were derived from the same patient), chemo-selected cells were even more sensitive to cytotoxic lymphocytes, a difference not explained by a different distribution of neoplastic cells in the various cell cycle phases. We also studied the inhibitory activity of activated lymphocytes on the clonogenic potential of chemo-surviving tumor cells by the human tumor clonogenic assay. Inhibitions of tumor cell growth in the two patients tested were 100 and 94%, respectively; the activity of lymphocytes was dependent on the coculture time and the effector/target cell ratio. These data indicate that in vitro treatment with mafosfamide does not select cells resistant to the action of activated lymphocytes and that, given the right experimental conditions, these immune effectors can completely lyse tumor cells. PMID- 3032409 TI - Characterization of the cyclic adenosine 3':5'-monophosphate effector system in hormone-dependent and hormone-independent rat mammary carcinomas. AB - We have compared the properties of cyclic adenosine 3':5'-monophosphate (cAMP) dependent protein kinases I and II in hormone-dependent/cAMP-sensitive (DMBA tumor) and hormone-independent/cAMP-resistant (DMBA 1 tumor) rat mammary carcinomas. cAMP-resistance was not due to less total kinase in the hormone independent tumor, grossly altered distribution between soluble and particulate forms of the kinase (80% soluble in either tumor), alteration in the relative proportion of isozymes I and II of the protein kinase (the soluble and the particulate fraction from both tumors contained about 50% of either isozyme), or a decreased sensitivity towards cAMP (both isozymes had affinities for cAMP and its derivatives that corresponded closely with those of isozymes from normal tissues). Furthermore, the sensitivity of the enzymes towards thermal denaturation was identical for samples from the two tumor types. Subtle differences did, however, exist between the regulatory moieties [regulatory subunit of cAMP-dependent protein kinase II (RII)] of isozyme II from the two tumors: autophosphorylated RII from the hormone-independent tumor migrated as a doublet corresponding to Mrs 54,000 and 52,000 on sodium dodecyl sulfate polyacrylamide gels, against Mrs 53,000 and 52,000 for RII from the hormone dependent tumor; RII from the two tumors showed different elution profiles upon DEAE-cellulose chromatography; a considerable proportion of the soluble RII in the hormone-independent tumor formed supramolecular aggregates as judged by size exclusion chromatography. No such microheterogeneity was noted for isozyme I. This study thus shows that the lack of cAMP-responsiveness of one tumor is related either to a defect distal to the cAMP-dependent protein kinases or to the appearance of the new subtype of RII in the resistant tumor. If the latter explanation is correct, it means that the part of the RII molecule responsible for interaction with other proteins rather than that responsible for cAMP-binding and control of protein kinase activity modulates the growth-inhibiting response to cAMP. PMID- 3032410 TI - Correlation of aromatic hydroxylation of 11 beta-substituted estrogens with morphological transformation in vitro but not with in vivo tumor induction by these hormones. AB - In estrogen-induced cancer, catechol formation from administered steroids has been postulated to be a necessary event for estrogen activation and subsequent damage to cellular macromolecules. In the present study, this hypothesis has been tested using two homologous series of structurally related estrogens: estradiol, 11 beta-methylestradiol, 11 beta-ethylestradiol, 11 beta-methyl-17 alpha ethinylestradiol, 11 beta-ethyl-17 alpha-ethinylestradiol, 11 beta-methoxy-17 alpha-ethinylestradiol, and 17 alpha-ethinylestradiol. In the Syrian hamster renal carcinoma model, only 11 beta-methylestradiol and 17 alpha-ethinylestradiol were weak carcinogens (2 of 20 and 2 of 24 hamsters with tumors, respectively). The other estrogens tested induced renal carcinoma within 6 to 9 months with an incidence in the 80-100% range. The tumor incidence in vivo did not correlate with the rates of catechol formation by hamster kidney microsomes in vitro. Compared to estradiol (relative rate, 100), catechol formation by the substituted estrogens was significantly lower, ranging from 48 (11 beta-methylestradiol) to 2 (11 beta-methoxy-17 alpha-ethinylestradiol). Kidney DNA of hamsters treated with the four 17 alpha-ethinyl estrogens, when analyzed by 32P postlabeling assay, contained the same set of covalently modified nucleotides the formation of which had previously been found to precede estrogen-induced renal carcinogenesis in vivo. In contrast, relative rates of catechol estrogen formation by BALB/c 3T3 microsomes correlated with induction of morphological transformation of BALB/c 3T3 cells and decreased in the following order: 11 beta-methylestradiol greater than 17 alpha-ethinylestradiol greater than or equal to estradiol greater than 11 beta-ethylestradiol greater than 11 beta-methoxy-17 alpha-ethinylestradiol. The hormonal potencies of several estrogen derivatives studied by various assays did not correlate with in vivo carcinogenic or in vitro cell-transforming activities. It is concluded from these experiments that in cell culture catechol formation and morphological transformation are directly related. In vivo, aromatic hydroxylation of administered estrogens did not correlate with the incidence of estrogen-induced renal carcinoma in Syrian hamsters. PMID- 3032411 TI - DNA-mediated transfer and cloning of a human multidrug-resistant gene of adriamycin-resistant myelogenous leukemia K562. AB - We have successfully transferred and cloned a fragment of a human multidrug resistant gene by using DNA-mediated gene transfer. Macromolecular DNA of human multidrug-resistant K562 cells was transfected to drug-sensitive mouse Ltk- cells to obtain a drug-resistant transfectant with a human resistant gene. Both primary and secondary transfectants showed similar patterns of cross-resistance to Adriamycin and vincristine. The mechanism of drug resistance of the transfectants was attributed to decreased retention of the drug. Three secondary transfectants obtained independently contained common Alu-containing EcoRI fragments 15, 6.5, 3.7, 2.6, and 1.9 kilobases long. The 2.6-kilobase EcoRI fragment was cloned from a lambda phage genomic library made from DNA of a secondary transfectant. The 2.6 kilobase fragment was detected in the primary and secondary transfectants but not in the parental Ltk-, Adriamycin-resistant Ltk-, and Adriamycin-resistant P388 cells. This sequence was found to be amplified in several multidrug-resistant cell lines such as Adriamycin-resistant ovarian carcinoma A2780 and colchicine resistant KB carcinoma cells. The 2.6-kilobase fragment hybridized with a 4.5 kilobase mRNA which is overexpressed in the Adriamycin-resistant K562 cells and the Adriamycin-resistant A2780 cells but not detected in the parental K562 cells. The gene transferred and cloned in this study seems to be related to the P glycoprotein gene as judged from the size of mRNA and its overexpression in some of the multidrug-resistant cell lines where P-glycoprotein was found to be highly expressed. PMID- 3032412 TI - Molecular evidence for the lack of epidermal growth factor receptor gene expression in small cell lung carcinoma cells. AB - It has been shown that none of the small cell lung carcinoma (SCLC) cell lines possess epidermal growth factor (EGF) binding activity on their surface. We have examined several SCLC cell lines for the possibility that they may have EGF receptors but that the receptors are masked by an EGF-like protein factor(s), which may be produced by an autocrine mechanism. No evidence, however, was found for the production of such factors. We then used an EGF receptor complementary DNA to determine the state of the EGF receptor gene by Southern blot analysis. The receptor gene appears to be present in these cells in an intact, unrearranged form. These cells, however, were found to lack detectable levels of EGF receptor mRNA, suggesting a possible reason for the absence of EGF receptors on the cell surface. Furthermore, karyotype analysis revealed that SCLC cell lines Lu134 and H69 contained a morphologically normal chromosome 7, which carries the EGF receptor gene. Also, these SCLC cells contained the apparently normal chromosome 3 and exhibited the presence of c-raf-1 gene in an unrearranged form. Thus, the previously noted partial deletion of chromosome 3 is not necessarily common to the SCLC cells. Instead, the lack of EGF receptor is frequently found in SCLC cell lines and is distinct from the other types of lung cancer. We postulate that SCLC cells have some active regulatory mechanism which prevents the expression of EGF receptor gene. PMID- 3032413 TI - Determinants of complete remission induction and maintenance in chemotherapy with or without irradiation of small cell lung cancer. AB - The possible influence of pretreatment patient characteristics upon the probabilities of complete remission (CR) induction and maintenance was investigated in a series of 815 nonresected patients with small cell lung cancer. All patients underwent pretreatment staging which enabled allocation of 391 patients to trials for limited stage disease and 424 patients to trials for extensive disease. Three controlled trials for each disease stage were conducted between 1973 and 1981. All therapeutic regimens consisted of combinations of between three and six agents (lomustine, cyclophosphamide, methotrexate, vincristine, doxorubicin, etoposide) with or without irradiation. Thirty-five % of the limited stage patients and 18% of the extensive stage patients were alive and had achieved a complete remission 16 weeks after initiation of the treatment, i.e., after four cycles of chemotherapy. Relationships between pretreatment characteristics and the probability to pass this benchmark were examined by logistic regression analysis. The probability of CR was negatively related to increased serum lactate dehydrogenase and male sex in both disease stages. Pretreatment anemia (less than 12 g/liter) and poor performance status were associated with a reduced CR rate in limited and extensive stage disease, respectively. Factors related to the maintenance of complete remission were subsequently examined in the 211 complete responders by use of Cox's regression analysis. Complete responders with extensive disease prior to treatment had greater cumulative risk of relapse than those with limited disease (P less than 0.01). Hyponatremia had a significant negative influence on the remission duration in limited disease while age greater than 60 years and bone marrow metastases had significantly negative influence in extensive disease. Using the models it was possible to identify subgroups of patients with CR rates ranging from 5 to 55% and to stratify complete responders according to estimated risks of subsequent relapse. PMID- 3032414 TI - Detection of small cell lung cancer bone marrow involvement by discontinuous gradient sedimentation. AB - Marrow involvement by small cell lung cancer (SCLC) is detected in 10-23% of patients at initial diagnosis by marrow aspirate and biopsy techniques. To improve the detection and potentially monitor marrow involvement by SCLC we have attempted to concentrate malignant cells with clonogenic potential on a discontinuous density gradient (DDG). The bone marrow from 43 patients with SCLC (36 with histologically negative marrow aspirates and biopsies) were separated on a Ficoll-based DDG. Samples were also separated by conventional Ficoll diatrizoate (FD) (density, 1.077) gradient sedimentation. The cellular interphase from three fractions (F X) corresponding to specific densities 1.050 (F X 1), 1.055 (F X 2), and 1.060 (F X 3) as well as cells separated by Ficoll-diatrizoate (F X FD) centrifugation were isolated and 2.5 X 10(5) cells from each fraction were cultured in 2 ml of 0.3% agar in McCoy's media with 10% fetal calf serum, 2.5 micrograms transferrin, 1 microgram insulin, and 1% penicillin-streptomycin. Colony growth was assessed after 14 days of culture at 37 degrees C and 6% CO2. Tumor colony growth was seen in eight of 36 (22%) patients with histologically negative marrows as well as in four of seven (57%) patients with known involvement. Mean colony growth per 2.5 X 10(5) cells for all 12 patients was 4.3 colonies for F X 1; 8.8 for F X 2; and 2.7 for F X 3. In contrast mean growth from the F X FD was 1.0 colonies. Cells with clonogenic potential could be demonstrated from F X 2 and F X 3 in seven of 12 and eight of 12 patients, respectively; in F X FD four of 12 patients had tumor growth. We conclude that separation of marrow samples by DDG concentrates malignant cells with clonogenic potential at least 8-fold compared to FD separation and that the sensitivity of the clonogenic assay in detecting marrow involvement by SCLC is enhanced by DDG sedimentation. PMID- 3032415 TI - Inhibition of human erythrocyte inositol lipid metabolism by adriamycin. AB - Incubation of human erythrocytes with Adriamycin prevented their morphological transition from discocytes to echinocytes when they were either depleted of ATP or loaded with calcium. This effect was dependent upon drug concentration and cell density. Adriamycin (10(-5) M) prevented, by greater than 90%, the echinocytosis of 10(7) cells/ml (S. B. Chahwala and J. A. Hickman, Cancer Res., 45: 4986-4989, 1985), and 5 X 10(-4) M prevented that of 10(9) cells/ml. There was a poor correlation between the effects of Adriamycin as a modulator of this morphological transition and its potency as an inhibitor of calmodulin. Using inside-out red blood cell vesicles, Adriamycin inhibited calmodulin dependent Ca2+ uptake with a 50% inhibitory concentration of 5 X 10(-4) M. Adriamycin thus differs from other amphipathic drugs, such as those of the phenothiazine class, where inhibition of calmodulin correlated well with effects on erythrocyte morphology (G. A. Nelson, M. L. Andrews, and M. J. Karnovsky, J. Cell Biol., 96: 730-735, 1983). After 12 h of ATP depletion, levels of phosphatidylinositol 4,5 bisphosphate [PtdIns(4,5)P2] extracted from 10(9) erythrocytes/ml fell by 57% and after 48 h they fell by 97%, changes which were coincident with a 100% transition of morphology to echinocytes. Adriamycin, 5 X 10(-4) M-1 X 10(-3) M, maintained 10(9) cells/ml in a discocyte morphology and maintained PtdIns(4,5)P2 levels at 60-70% of the time zero controls, independently of the size of the fall in PtdIns(4,5)P2 levels. The data suggested that Adriamycin inhibited a discrete pool of PtdIns(4,5)P2 which may be responsible for the maintenance of a discocyte morphology. Neomycin, 10(-3) M, had no effect on the ATP depletion-induced discocyte-echinocyte transition of 10(9) erythrocytes/ml or on the fall in PtdIns(4,5)P2 levels. Adriamycin, like neomycin, prevented the calcium-induced breakdown of erythrocyte membrane vesicle PtdIns(4,5)P2 to inositol trisphosphate (50% inhibitory concentration, 7 X 10(-4) M) but, unlike neomycin (50% inhibitory concentration, 4.25 X 10(-4) M) it was able to inhibit breakdown by 100% at higher concentrations. PMID- 3032416 TI - Detection of human lung epithelial cell growth factors produced by a lung carcinoma cell line: use in culture of primary solid lung tumors. AB - Serum-free medium conditioned for 72 h by a human bronchioloalveolar carcinoma of lung, A549-1, stimulated the colony formation of normal human bronchial epithelial cells, newly cultured cells from human solid lung tumors, and established human lung tumor cell lines, including A549-1 cells themselves. This activity was concentration dependent and was stable to acid. Growth factors in A549-1 conditioned medium (CM) supported culture of solid lung tumors; primary cell cultures were obtained from nine of 10 solid lung tumors of non-small cell origin and from one small cell tumor using A549-1 CM. In addition, three cell lines have been established to date from these primary cultures. Gel filtration of concentrated A549-1 CM on Biogel P-10 separated the growth promoting activity into four regions of apparent Mr 70,000, 12,000, 8,000, and 6,000, and two broad regions of apparent Mr 3000-5000. All but the 12,000 Mr fraction contained activity which competed for specific binding of epidermal growth factor (EGF) to A431 cell membranes. CM was superior to both EGF and TGF alpha in stimulating growth of normal and neoplastic lung cells. EGF also was inhibitory to tumor cells while TGF alpha stimulated both normal and tumor cell growth. TGF beta was also found in CM but inhibited normal and neoplastic lung epithelial cell growth. Of other substances tested, ILGF-I stimulated colony formation. The results suggest that autocrine factors may be important in non-small cell lung tumor cell growth and that differences in response to EGF and TGF alpha may provide the basis for selective culturing of normal and neoplastic lung epithelial cells. PMID- 3032417 TI - Differential effects of steroid hormones on parameters of cell growth. AB - Steroid hormones affect the growth of many tumor cells both in vivo and in vitro. Growth of cells in vitro can be studied as either anchorage dependent (monolayer culture) or anchorage independent (suspension culture), and in each case, steroid hormones can alter log-phase proliferation rate, saturation density, or cell morphology. Results presented here demonstrate that different steroid hormones can have different effects on each of these parameters in breast cancer cells. In androgen-responsive cells, glucocorticoid and androgen both stimulated fibroblastic morphology and saturation density in monolayer and growth in suspension, but glucocorticoid inhibited log-phase proliferation rate while androgen stimulated it. By transfection experiments, it has been possible to separate the androgen-regulated stimulation of cell proliferation from the androgen-regulated cell morphology changes indicating that the two parameters are not totally interdependent. The same separation, however, was not achieved for glucocorticoid regulation. Further separation of hormone effects was achieved also during the natural course of progression to steroid autonomy. In particular, the androgen and glucocorticoid stimulations of growth in suspension were separated. The experiments described here carry important messages for the design and interpretation of any experiment aimed at elucidating molecular events involved in steroid-mediated cell growth in culture. PMID- 3032418 TI - Gestational risk factors for Wilms' tumor: results of a case-control study. AB - Gestational risk factors for Wilms' tumor were investigated in a pair-matched case-control study. Cases who were under 15 years of age at diagnosis during 1970 1983 were identified through the registries of the three main hospitals treating childhood cancer in the greater Philadelphia area. Controls were selected by random digit dialing and were matched to cases on race, birth date, and telephone area code and exchange. Because of a low participation rate among nonwhites, results are reported only for the 88 white matched pairs whose parents were interviewed by telephone. Of the hypothesized risk factors, maternal use of hair coloring products in the year prior to the index child's birth (odds ratio, 3.6; P = 0.003) and hypertension or fluid retention during pregnancy (odds ratio, 5.0, P = 0.01) were significantly associated with increased risk of Wilms' tumor. Use of hair-coloring products was strongly associated with cases in which Wilms' tumor was diagnosed before 2 years of age (odds ratio, 15; P = 0.001). For two other gestational factors, tea drinking and vaginal infection, the odds ratios were significantly elevated for all cases and the effects were concentrated among the nongenetic cases. Bilateral cases had a significantly higher mean birth weight than did their controls. Older maternal age was significantly associated with the genetic form of Wilms' tumor. Adjustment for possible confounders and consideration of the time interval between the index pregnancy and the interview did not substantially alter the findings. PMID- 3032419 TI - Stimulation of the respiratory burst of murine peritoneal inflammatory neutrophils by conjugation with tumor cells. AB - Murine peritoneal neutrophils (PMNs), elicited by i.p. injection of formalin killed Corynebacteria parvum, spontaneously lyse teratocarcinoma targets through the secretion of reactive oxygen intermediates. Examination of effector-target interactions at the single cell level revealed that PMNs conjugated to tumor cells were 3-fold more frequently stained by nitroblue tetrazolium compared to nonconjugating PMNs suggesting that tumor targets stimulated a potent tumor-lytic respiratory burst. This notion was confirmed by the detection of superoxide and hydrogen peroxide generation from PMNs as well as a luminol-dependent chemiluminescent response following conjugation with viable tumor targets. Generation of superoxide was dependent upon the presence of dihydrocytochalasin B. In addition to teratocarcinoma cells, comparable stimulation was achieved by conjugation with YAC and P815 targets but not thymocytes. Reactive oxygen intermediate release was also achieved by mixing peritoneal PMNs with heat-killed tumor cells. In contrast to bacteria-induced effectors, PMNs elicited by i.p. injection of thioglycollate were incapable of responding following conjugation with tumor targets although they were competent for reactive oxygen intermediate release when stimulated by phorbol myristate acetate. Teratocarcinoma targets were sensitive to concentrations of H2O2 that could be achieved by PMNs following contact. These data indicate that Corynebacteria-elicited inflammatory PMNs lyse their bound tumor targets by a mechanism similar to a stimulus-secretion model. PMID- 3032420 TI - Molecular properties of F9 embryoglycan recognized by a unique antibody in sera from patients with germ cell tumors. AB - Human antibody against an embryoglycan present on a mouse teratocarcinoma cell line F9 was found in sera from 16 of 29 patients with embryonal carcinoma, yolk sac tumor, immature teratoma, and choriocarcinoma of gonadal and extragonadal origins by Farr assay. In contrast, none of the sera from patients (77 cases) with dysgerminoma, seminoma, germinoma, and mature teratoma or from patients (118 cases) with nongerm cell types of ovarian tumors contained this antibody. The antigenic embryoglycan was of high molecular weight (Mr greater than 70,000) on Sephacryl S300 column chromatography and carried binding sites for Grifonia simplicifolia agglutinin-1. The antigenic embryoglycan was also found in F9 cell cultured medium. Absorption of patients' sera with synthetic Blood Group B trisaccharides failed to remove the antibody against F9 embryoglycan. None of these patients' sera showed higher hemagglutination titer to rabbit erythrocytes than the normal range. In contrast, alpha-galactosyl carbohydrates obtained from Ehrlich ascites tumor cells effectively inhibited the binding of patients' sera with F9 embryoglycan. These results indicate that the human antibody against F9 embryoglycan recognizes alpha-galactosyl structures that are distinct from B blood group antigen, but are cross-reactive with alpha-galactosyl structures on Ehrlich ascites cells. PMID- 3032422 TI - Differential effects of dibutyryl cyclic adenosine monophosphate and retinoic acid on the growth, differentiation, and cyclic adenosine monophosphate-binding protein of murine neuroblastoma cells. AB - Dibutyryl cyclic adenosine 3':5'-monophosphate (Bt2cAMP) and beta-all-trans retinoic acid (RA) have been shown separately, and in some cases in combination, to modulate the growth, differentiation, and cAMP-dependent protein kinase (PK-A) activity of various tumor cells. The effects of Bt2cAMP and RA on a cholinergic clone (S20) of C1300 mouse neuroblastoma cells were explored in the present study. Treatment of these cells with 1 mM Bt2cAMP for 3 or more days resulted in 93% inhibition of cell proliferation in monolayer cultures and in 98% inhibition of colony formation in semisolid medium (0.5% agarose). In contrast, treatment of the cells with 1 or 10 microM RA had no inhibitory effects on cell proliferation in monolayer cultures but enhanced colony formation in agarose by up to 130%. The growth of cells treated with a combination of Bt2cAMP and RA was inhibited, although less so than with Bt2cAMP alone. Cells treated with Bt2cAMP alone or Bt2cAMP and RA extended long, neurite-like, cellular processes indicative of differentiation, whereas only a few untreated or RA-treated cells produced such extensions. The amount of [3H]cAMP-binding protein increased gradually up to 2 fold during a 3-day treatment with Bt2cAMP; in contrast it decreased by nearly 2 fold during RA treatment. These changes occurred in the level of the type I regulatory subunit (RI) of PK-A as determined by photoaffinity labeling with 8 azidoadenosine cyclic 3':5'-[32P]monophosphate. The increase in RI following Bt2cAMP treatment was corroborated by DEAE-cellulose chromatography. This analysis also demonstrated that type I PK-A is the predominant kinase in the untreated S20 cells and that RI exists as a free subunit in Bt2cAMP-treated cells. The activity of PK-A decreased by about 20% following treatment with either Bt2cAMP or RA and by 45% following treatment with a combination of both agents. These results suggest that the distinct effects of Bt2cAMP and RA on the anchorage-independent growth of S20 cells may be related to their opposite effects on the level of RI. PMID- 3032421 TI - Role of metabolism and oxidation-reduction cycling in the cytotoxicity of antitumor quinoneimines and quinonediimines. AB - Quinone(di)imines are nitrogen analogues of quinones in which one or both quinone oxygens are replaced by an imino group. A series of quinone(di)imines with antitumor activity has been studied for its in vitro chemical reactivity, metabolism, acute toxicity to primary cultured rat hepatocytes, and growth inhibitory activity with Chinese hamster ovary (CHO) cells. The quinone(di)imines exhibited a wide range of activity as substrates for metabolism by hepatic microsomal flavoenzymes. The maximum rate of quinone(di)imine metabolism was more than 7.5-fold greater than reported for metabolism of quinones. Some quinone(di)imines formed free radicals that could be detected by electron spin resonance spectroscopy. 2-Amino-1,4-naphthoquinoneimine gave a short-lived electron spin resonance signal that could be detected only under aerobic conditions. 2,3',6-Trichloroindophenol gave an electron spin resonance signal in air that was stable for 24 h. Most quinone(di)imines underwent oxidation reduction cycling to form the superoxide anion radical, but some quinone(di)imines, although rapidly metabolized, formed little or no superoxide anion radical. Quinone(di)imines were relatively toxic to hepatocytes and CHO cells, and some quinone(di)imines were more toxic to one cell type than the other. The log 1-octanol/water partition coefficient showed an optimal value of 2.61 for toxicity against both cell types. In hepatocytes the more toxic quinone(di)imines were the most rapidly metabolized. For a subgroup of quinone(di)imines toxicity to hepatocytes and CHO cells appeared to be related to the ability to form a semiquinone(di)imine free radical. Toxicity of quinone(di)imines to hepatocytes and CHO cells was not related to superoxide anion radical formation, and toxicity to CHO cells was not affected by exclusion of oxygen during exposure of the cells to the compounds. The rate of chemical addition of quinone(di)imines to reduced glutathione did not correlate with toxicity. An understanding of the mechanisms of acute toxicity and growth inhibitory activity of quinone(di)imines could lead to the design of more selective quinonoid antitumor agents. PMID- 3032424 TI - Interactions between catecholamines and their receptors in blood. PMID- 3032423 TI - Quantitative assay of human T-cell leukemia/lymphoma virus transformation. AB - The in vitro transformation of normal T-lymphocytes by human T-cell leukemia/lymphoma virus (HTLV-I) is possible utilizing cocultivation techniques. We now report on a quantitative assay for HTLV-I transformation. Transformed cell lines were produced by cocultivation of either preactivated (phytohemagglutinin and T-cell growth factor) or nonactivated peripheral blood mononuclear cells with an equal number of lethally irradiated HTLV-I-positive donor cells (MT-2). After 14 days in liquid culture, transformed cells were plated in a 2-layer soft agarose system with or without T-cell growth factor (TCGF). Colony formation among 50 normal controls was observed at varying efficiencies with a mean number of 179 colonies (range, 6-599) in the presence of TCGF (up to a 2-log difference). The day 14 T-cell cultures demonstrated relatively low colony forming efficiencies (less than or equal to 0.1%) and enhanced colony formation in the presence of TCGF. Day 14 after cocultivation was chosen for this assay based on a dose-response relationship between colony formation and the virus positive donor cell inoculum and the known kinetics of colony growth of normal activated T-cells. An analysis of individual colonies indicated that they were of target cell origin and HTLV-I positive. Recombinant beta-interferon in increasing concentrations caused a decrease in colony formation as measured in this assay. Long-term cell cultures (2-18 months) showed higher colony-forming efficiencies (up to 1.0%) which were not enhanced by TCGF. The ability to quantitatively evaluate transformation via colony counts will provide an opportunity to study differences in transforming efficiencies attributable to varying target cells, donor cells, or blocking factors such as interferons, drugs, or anti-HTLV-I antibodies. PMID- 3032425 TI - Second messengers, natural rewards, and drugs of abuse. PMID- 3032426 TI - Structure-activity relationship of peptide fragments derived from DBI (diazepam binding inhibitor), a putative endogenous ligand of benzodiazepine recognition sites. PMID- 3032427 TI - Endogenous modulator for nitrendipine binding sites. PMID- 3032428 TI - Adenylate cyclase coupled beta-adrenergic receptors: biochemical mechanisms of desensitization. PMID- 3032429 TI - Gangliosides reduce mortality due to global ischemia: membrane protection. PMID- 3032430 TI - Blood platelets as neuronal models: use and limitations. PMID- 3032431 TI - Biochemical aspects of suicide. PMID- 3032432 TI - The possible role of locus coeruleus noradrenergic activity in anxiety-panic. PMID- 3032433 TI - The benzodiazepine/GABA receptor complex in anxiety. PMID- 3032435 TI - Meningioma preceding breast cancer. PMID- 3032434 TI - High-dose etoposide therapy for extensive small cell lung cancer: a Cancer and Leukemia Group B Study. AB - Twenty-one previously untreated patients with extensive small cell lung cancer were treated with etoposide, 400 mg/m2/day for 3 consecutive days. Myelosuppression was severe, with a treatment-related death rate of 28%. Five partial responses were achieved. High-dose etoposide as given in this study produced unacceptable toxicity and no complete responses. PMID- 3032436 TI - Effects of ascorbate ions on intracellular fluorescein emission polarization spectra in cancer and normal proliferating cells. AB - Ascorbate ions induced a polarization peak in the intracellular fluorescein fluorescence polarization spectra of asynchronous populations of a variety of cancer cell types from human and animal biopsy material or in cells grown in vivo and/or in vitro. The appearance of this polarization peak at 510 nm on excitation at 470 nm (P510 peak) indicates the transition of mitochondria from the condensed (active) into orthodox (resting) conformation. In contrast, no effects of ascorbate ions were observed in the polarization spectra of various normal cell lines. This apparent differential response seems to be caused by the effects of ascorbate ions on several steps of the altered energy metabolism in cancer cells. It appeared not to be due to a defective mechanism responsible for the conformational changes in the mitochondria. In YMDR cells resistant to the antitumor drug methylene-dimethanesulphonate (MDMS), HeLa cells pretreated with colcemid, and YMDS cells pretreated with cytochalasin B, the transition of mitochondria into orthodox conformation could not be induced by ascorbate ions. Thus, it is possible that tumors also pretreated with other drugs become resistant to growth inhibitory effects of ascorbate ions. Induction of the fluorescein emission polarization peak at 510 nm in cells from biopsies or from in vitro-induced malignancies could be used as an additional criterion for malignant transformation. PMID- 3032437 TI - Role of cellular retinoic acid binding protein (cRABP) in patients with large bowel cancer. AB - The presence of cellular retinoic acid binding protein (cRABP) was analyzed in 13 consecutive patients with large bowel cancer (one right colon and 12 rectum sigmoid, classified as three Duke B, eight C, and two D). Specimens of neoplastic tissue and of adjacent mucosa were obtained at surgery, and cRABP was determined by an assay based on incubation of partially purified cytosol with labeled retinoic acid and ultracentrifugation in sucrose density gradients. Sixty-one percent of tumors contained detectable levels of cRABP, whereas 58.3% of normal mucosal specimens were positive for cRABP. Among the positive tumors 62.5% contained cRABP also in the corresponding mucosa; in the group of cRABP-negative tumors, 40% showed cRABP in the adjacent mucosa. No correlation could be established with the grading or the stage of the tumors; however, interestingly, 100% (three cases) of gelatinous carcinomas were cRABP positive. Since cRABP seems to be a character of neoplastic cells contrary to normal ones, it would be interesting to investigate the conditions that influence the presence of this protein in normal appearing mucosa adjacent to carcinoma. PMID- 3032438 TI - Estimation of poly (C) avid serum ribonuclease in hepatoma patients. AB - Human poly (C) avid serum ribonuclease (RNase) differs in physico-chemical, electrophoretic, and catalytic properties from ribonuclease activity encountered in liver preparations. The first is reported as "secretory type", the latter, because it is undetectable in body fluids, as "nonsecretory type". We determined RNase activity in 11 hepatoma patients. A statistical difference from a normal control of corresponding age was encountered in both age groups investigated (51 60 years, P less than 0.05; 61-70 years, P less than 0.01). The circumstances mentioned above make the tumor itself unlikely to be the source of RNase elevation. Besides a diminished synthesis of RNase inhibitor by hepatoma cells, tumor-derived polyamines could contribute to enhanced RNase activity. The influence of polyamines on RNase activity has already been demonstrated by in vitro experiments. Simultaneous estimation of polyamines and RNase is required to elucidate in vivo circumstances. PMID- 3032439 TI - Rapid diagnosis of liver cancer by ultrasound-guided fine-needle aspiration biopsy. AB - Because of the relatively favorable prognosis to the patient with early detected hepatocarcinoma followed by surgical treatment if resection is possible, it is important to differentiate quickly between primary and secondary liver cancer. Ultrasound-guided percutaneous fine needle aspiration biopsy (US-FNAB) was used as a first diagnostic measure in patients with sonographic evidence of liver tumors. Biopsies were done under sonographic control and antiseptic conditions from the center and the border zone of solid tumors of the liver, and the aspirated cell material was air dried on glass slides and Giemsa stained. The cytologic diagnosis was proved by clinical course and in most cases by surgical or autoptic histology. Cytologic evaluation lead in 15 cases to the diagnosis of definitive or suspicious malignant liver disease; the sensitivity was 93% and the specificity was 87%. One case classified as suspicious for malignancy by cytologic examination could be identified as cirrhotic nodule by further investigations. In none of the patients did we find complications from the biopsy procedure. From these data it is concluded that US-FNAB can serve as a rapid, inexpensive, safe, and highly accurate first diagnostic step in patients with solid lesions of the liver. PMID- 3032440 TI - Clearance of retroviremia and regression of malignancy in cats with leukemia lymphoma during treatment with staphylococcal protein A. AB - Cats persistently infected with a type C retrovirus (feline leukemia virus; FeLV) were treated either by the extracorporeal perfusion of their plasma through filter columns containing staphylococcal protein A (SPA) or by the intraperitoneal injection of SPA for up to 32 treatments. The clearance of evidence of viral infection occurred during treatment with SPA in 67% of the treated cats with chronic viral infection that had FeLV-related abnormalities other than overt malignancy. This rate of viral clearance is more than 30 times greater than the incidence of spontaneous clearance of persistent FeLV viremia in the untreated cat. Actual clearance of viremia occurred in 28% of all cats treated. Regression of malignancy, which occurred in four of 12 (33%) treated cats, was demonstrated by reductions in tumor size and bone marrow neoplastic cell populations. Immediately preceding or concurrent with these remissions from viral infection and malignant disease, a marked though transient elevation in the plasma level of circulating gamma-like interferon occurred in some responding cats and was followed by the appearance and rising titer of a complement dependent cytotoxic antibody that reacted with FeLV-infected cells. This IgG antibody was shown by several analyses to be specific for the major viral envelope glycoprotein gp70. Further investigations demonstrated that high levels of FeLV at the cellular level might be responsible for the reduced ability of normal feline lymphocytes to secrete gamma-interferon in response to mitogenic stimuli in vitro. These findings may provide some clue to the variation in the responsiveness of individual cats to immunotherapy with staphylococcal protein A. PMID- 3032441 TI - [Regulation of heart contraction: role of contractile proteins]. PMID- 3032442 TI - Anticalculus effect of a dentifrice containing pyrophosphate salts and sodium fluoride. PMID- 3032443 TI - Applications of hydroxylapatite in oral and maxillofacial surgery, part I: Periodontal and endosteal-implant repairs. PMID- 3032444 TI - The Keyes technique as a cofactor in self-inflicted gingival lesions: a case report. PMID- 3032445 TI - Cell-cell communication and hormonal imprinting: transmission of a receptor level effect of cells not directly influenced by the hormone. PMID- 3032448 TI - Transpososomes: stable protein-DNA complexes involved in the in vitro transposition of bacteriophage Mu DNA. AB - We report that two types of stable protein-DNA complexes, or transpososomes, are generated in vitro during the Mu DNA strand transfer reaction. The Type 1 complex is an intermediate in the reaction. Its formation requires a supercoiled mini-Mu donor plasmid, Mu A and HU protein, and Mg2+. In the Type 1 complex the two ends of Mu are held together, creating a figure eight-shaped molecule with two independent topological domains; the Mu sequences remain supercoiled while the vector DNA is relaxed because of nicking. In the presence of Mu B protein, ATP, target DNA, and Mg2+, the Type 1 complex is converted into the protein-associated product of the strand transfer reaction. In this Type 2 complex, the target DNA has been joined to the Mu DNA ends held in the synaptic complex at the center of the figure eight. Supercoils are not required for the latter reaction. PMID- 3032446 TI - Calmodulin blocker inhibits Ca++-ATPase activity in secretory ameloblast of rat incisor. AB - The effects of the calmodulin blocker, trifluoperazine (TEP), on membrane-bound Ca++-ATPase, Na+-K+-ATPase (EC 3.6.1.3.) and the ultrastructure of the enamel organ were investigated in the lower incisors of normal and TFP-injected rats. The rats, of about 100 g body weight, were given either 0.2 ml physiological saline or 100 micrograms TFP dissolved in 0.2 ml physiological saline through a jugular vein and fixed by transcardiac perfusion with a formaldehyde glutaraldehyde mixture at 1 and 2 h after TFP administration. Non-decalcified sections of the enamel organ less than 50 micron in thickness, prepared from dissected lower incisors, were processed for the ultracytochemical demonstration of Ca++-ATPase and Na+-K+-ATPase by the one-step lead method at alkaline pH. In control saline-injected animals the most intense enzymatic reaction of Ca++ ATPase was demonstrated along the plasma membranes of the entire cell surfaces of secretory ameloblasts. Moderate enzymatic reaction was also observed in the plasma membranes of the cells of stratum intermedium and papillary layer. Reaction precipitates of Na+-K+-ATPase activity were localized clearly along the plasma membranes of only the cells of stratum intermedium and papillary layer. The most drastic effect of TFP was a marked disappearance of enzymatic reaction of Ca++-ATPase from the plasma membranes of secretory ameloblasts, except for a weak persistent reaction in the basolateral cell surfaces of the infranuclear region facing the stratum intermedium. The cells of stratum intermedium and papillary layer, however, continued to react for Ca++-ATPase even after TFP treatment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3032447 TI - Pathogenesis of Burkitt lymphoma: expression of an activated c-myc oncogene causes the tumorigenic conversion of EBV-infected human B lymphoblasts. AB - To study the pathogenesis of Burkitt lymphoma, we introduced activated c-myc genes into human EBV-infected lymphoblastoid cells derived from in vitro infection of normal cord blood or directly from infected peripheral blood from AIDS patients. In both cell types the constitutive expression of exogenous c-myc caused negative regulation of endogenous c-myc expression, changes in growth properties typical of transformed cells, and acquisition of tumorigenicity in immunodeficient mice. In all myc-transfected populations the degree of malignancy directly correlated with the level of c-myc mRNA. EBV infection and c-myc activation are thus sufficient for the tumorigenic conversion of human B cells in vitro, strongly supporting the hypothesis that these same two pathogenetic steps may be involved in the in vivo development of Burkitt lymphoma. PMID- 3032449 TI - Transformation by Rous sarcoma virus induces a novel gene with homology to a mitogenic platelet protein. AB - A cDNA clone, designated 9E3, was isolated from a chick embryo fibroblast (CEF) cDNA library. 9E3 mRNA was 20-fold higher in CEF following transformation by Rous sarcoma virus because of increased transcription rate. In CEF infected with temperature-sensitive mutants, increased 9E3 mRNA was found within 2 hr of a shift to permissive temperature. Nucleotide sequence and in vitro translation results indicate that 9E3 mRNA encodes an 11 kd polypeptide that is homologous to human connective tissue activating peptide III (CTAP-III), a mitogenic platelet alpha-granule protein, and to beta-thromboglobulin and platelet factor 4. The reported biological activities of CTAP-III suggest that elevated expression of 9E3 may play a role in producing some of the phenotypic features of RSV transformed cells. PMID- 3032450 TI - Correct integration of retroviral DNA in vitro. AB - We have developed a cell-free system for studying the integration of retroviral DNA. In our assay, amber mutations in a bacteriophage lambda genome that serves as the target for integration are suppressed by integration of an MLV derivative that carries the E. coli supF gene. The structure of the reaction products is that expected from an authentic MLV integration reaction. Linear viral DNA from the cytoplasm of infected cells serves as a precursor, though not necessarily the immediate precursor, to the provirus integrated in vitro. The viral DNA in the infected cell appears to be tightly associated with the enzymatic machinery required for its integration. Supercoiling, chromatin structure, transcription, and replication are not required of the target DNA. Since no high-energy cofactor is necessary, the DNA breakage and joining steps in the integration reaction are probably coupled. PMID- 3032451 TI - Interferon-beta gene regulation: tandemly repeated sequences of a synthetic 6 bp oligomer function as a virus-inducible enhancer. AB - Evidence is presented that virus-induced transcriptional activation of the human interferon-beta gene is mediated by repeated sequence units each consisting of 6 bp. The repeated units are present in the region between -65 and -109 relative to the cap site. We chemically synthesized the "consensus" DNA sequence units and tested their ability to confer virus inducibility on cognate and noncognate promoters. Most of such sequence units when tandemly repeated indeed mediate virus-induced activation of the promoter sequences. The most efficient sequence unit, AAGTGA, contributes incrementally in virus-induced activation of transcription and manifests properties of an inducible enhancer. Evidence is also provided that suggests that interaction of a positive regulatory factor(s) with this regulatory region. PMID- 3032452 TI - Recombination at the human alpha-globin gene cluster: sequence features and topological constraints. AB - We have characterized 170 kb of DNA around the human alpha-globin gene cluster to enable a systematic analysis of 12 naturally occurring deletions from this region. In 8 deletions, the 3' breakpoints lie within a 6-8 kb segment of DNA, identifying a breakpoint cluster region. Members of the Alu family of repetitive sequences are frequently found at the breakpoints and we describe a novel deletion due to homologous recombination between such repeats. In another deletion the breakpoints are separated by 131 bp of DNA, which we have shown to be transposed from a region 36 kb upstream from the 5' breakpoint where it is present in the inverse orientation. The sizes of these deletions, the nonrandom distribution of their breakpoints, and the nature of the inversion-duplication transposition event suggest that these rearrangements are constrained by the higher-order structure of the alpha-globin cluster. PMID- 3032453 TI - Phosphorylation within a specific domain of the phosphoprotein of vesicular stomatitis virus regulates transcription in vitro. AB - We have investigated the functional significance of phosphoserine residues that lie in the L protein-binding domain between amino acids 213 and 247 of the phosphoprotein (NS) of vesicular stomatitis virus. A series of mutant NS proteins were made by cell-free translation of mRNAs transcribed from the cloned gene. Site-directed substitution of alanine for both serine 236 and serine 242 essentially abolished RNA synthesis catalyzed by the NS-L complex. Substitution of either of these serines reduced RNA synthesis by 75%. Serine 218 played no major role in RNA synthesis. Phosphorylation of NS by the L protein was abrogated by substitution of either serine 236 or serine 242. These results indicate that phosphorylation of serines 236 and 242 in the NS protein regulates its binding with the L protein and the N-RNA template and is essential for activation of viral RNA synthesis. PMID- 3032454 TI - Cloning and expression of steroid sulfatase cDNA and the frequent occurrence of deletions in STS deficiency: implications for X-Y interchange. AB - Human STS is a microsomal enzyme important in steroid metabolism. The gene encoding STS is pseudoautosomal in the mouse but not in humans, and escapes X inactivation in both species. We have prepared monoclonal and polyclonal antibodies to the protein which has been purified and from which partial amino acid sequence data have been obtained. cDNA clones containing the entire coding sequence were isolated, sequenced, and expressed in heterologous cells. Variable length transcripts have been shown to be present and due to usage of alternative poly(A) addition sites. The functional gene maps to Xp22.3-Xpter and there is a pseudogene on Yq suggesting a recent pericentric inversion. Absence of STS enzymatic activity occurs frequently in human populations and produces a visible phenotype of scaly skin or ichthyosis. Ten patients with inherited STS deficiency were studied and eight had complete gene deletions. The possibility that STS deficiency results from aberrant X-Y interchange is discussed. PMID- 3032455 TI - Molecular analysis of mbcl-2: structure and expression of the murine gene homologous to the human gene involved in follicular lymphoma. AB - We have cloned the mouse bcl-2 (mbcl-2) genomic locus and analyzed it in detail. The gene is comprised of two exons separated by more than 15 kb. Two species of mRNAs are produced, and DNA sequencing analysis shows that they code for two proteins differing at their C terminus: a 7.5 kb transcript codes for a polypeptide of 236 amino acids, mbcl-2 alpha, and a 2.4 kb transcript, which derives from the 5' exon only, codes for a protein of 199 amino acids, mbcl-2 beta. The gene is characterized by very long (5' about 1.4 kb, and 3' about 5.1 kb) untranslated regions surrounding the relatively short coding region. We have mapped the 5' end of the mbcl-2 mRNAs by S1 protection analysis, and we have analyzed the promoter region. The expression of the mbcl-2 gene was analyzed in different cell lines and in mouse tissues. Expression is tissue-specific in adult tissues: spleen and thymus express the highest level of mbcl-2 transcripts. The mbcl-2 gene maps to mouse chromosome 1. PMID- 3032457 TI - hobo is responsible for the induction of hybrid dysgenesis by strains of Drosophila melanogaster bearing the male recombination factor 23.5MRF. AB - The male recombination factor 23.5MRF, isolated ten years ago from a natural Greek population of Drosophila melanogaster, has been shown to induce hybrid dysgenesis when crossed to some M strains, in a fashion slightly different from that of most P strains. Furthermore, it was recently shown that 23.5MRF can also induce GD sterility when crossed to specific P strain females (e.g., Harwich, pi 2 and T-007). In these experiments, the P strains mentioned behaved like M strains in that they did not induce sterility in the reciprocal crosses involving 23.5MRF. We extended the analysis to show that 23.5MRF does not destabilize snW(M) and that a derivative with fewer full-length P elements behaves like an M strain toward the same P strains and still retains its dysgenic properties in the reciprocal crosses. We show that there is a strong correlation between the site of dysgenic chromosomal breakpoints induced by 23.5MRF and the localization of hobo elements on the second chromosome, and also that hobo elements are found associated with several 23.5MRF induced mutations. These results suggest that hobo elements are responsible for the aberrant dysgenic properties of this strain, and that they may express their dysgenic properties independent of the presence of P elements. PMID- 3032456 TI - Coexpression of MMTV/v-Ha-ras and MMTV/c-myc genes in transgenic mice: synergistic action of oncogenes in vivo. AB - We have derived and mated separate strains of transgenic mice that carry either the v-Ha-ras or the c-myc gene driven by the mouse mammary tumor virus (MMTV) promoter/enhancer. Mice carrying the MMTV/v-Ha-ras transgene manifest two distinct disturbances of cell growth. The first, a benign hyperplasia of the Harderian lacrimal gland, is diffuse, involves the entire gland, and likely requires only the abnormal action of the v-Ha-ras gene. The second involves the focal development of malignancies of mammary, salivary, and lymphoid tissue and likely requires additional somatic events. When the MMTV/v-Ha-ras and MMTV/c-myc strains are crossed to yield hybrid mice, their joint action results in a dramatic and synergistic acceleration of tumor formation. Since these tumors arise stochastically and are apparently monoclonal in origin, additional somatic events appear necessary for their full malignant progression, even in the presence of activated v-Ha-ras and c-myc transgenes. PMID- 3032458 TI - Mobilization of hobo elements residing within the decapentaplegic gene complex: suggestion of a new hybrid dysgenesis system in Drosophila melanogaster. AB - We present results demonstrating that the hobo family of transposable elements can promote high rates of chromosomal instability. Using strains with a hobo element inserted within the decapentaplegic gene complex (DPP-C), we have recovered numerous DPP-C mutations involving chromosomal rearrangements and deletions with one endpoint in the vicinity of the pre-existing hobo element. This hypermutability occurred in the germ lines of hybrid progeny from crosses involving strains containing hobo elements to strains lacking them. In some crosses, the offspring had rudimentary gonads, reminiscent of GD sterility. The germline hypermutability and infertility are similar to those produced by P element-mediated hybrid dysgenesis. Given the many genetic and molecular similarities of the P and hobo systems, we propose that a system analogous to P-M hybrid dysgenesis has been activated in the hobo+ X hobo- crosses. PMID- 3032459 TI - Negative regulation of mitosis by wee1+, a gene encoding a protein kinase homolog. AB - Fission yeast wee1- mutants initiate mitosis at half the cell size of wild type. The wee1+ activity is required to prevent lethal premature mitosis in cells that overproduce the mitotic inducer cdc25+. This lethal phenotype was used to clone wee1+ by complementation. When wee1+ expression is increased, mitosis is delayed until cells grow to a larger size. Thus wee1+ functions as a dose-dependent inhibitor of mitosis, the first such element to be specifically identified and cloned. The carboxy-terminal region of the predicted 112 kd wee1+ protein contains protein kinase consensus sequences, suggesting that negative regulation of mitosis involves protein phosphorylation. Genetic evidence indicates that wee1+ and cdc25+ compete in a control system regulating the cdc2+ protein kinase, which is required for mitotic initiation. PMID- 3032460 TI - Effects of tunicamycin on the differentiation of F9 cells induced by either retinoic acid or retinoic acid and dibutyryl cyclic AMP. AB - Tunicamycin (0.5 micrograms/ml) inhibited differentiation of F9 cells treated either with retinoic acid or with retinoic acid and dibutyryl cyclic AMP, as monitored by the activity of alkaline phosphatase and expression of cytokeratins. On the other hand, the pattern of the polysaccharide chain synthesis changed drastically with the treatment irrespective of the presence of tunicamycin. Therefore, phenotypes induced with retinoic acid are dissociated into two categories, one that is directly induced by the drug and the other that is induced indirectly by a mechanism in which glycoproteins play a role. PMID- 3032462 TI - Differentiation of retrovirus-infected avian neuroretina cells. AB - The effects of oncogenic retroviruses on the expression of differentiation markers were studied in monolayer cultures of chick and quail embryo neuroretinas. Transformation by Rous sarcoma virus (RSV) did not affect the appearance of synapses, and the expression of glutamic acid decarboxylase was stimulated by pp60v-src, the product of the src gene. Quail embryo neuroretina cells transformed by Mill Hill 2 (which contains the two oncogenes v-mil and v myc) were induced to proliferate into permanent cultures that synthesized crystallins and produced lentoid bodies. In contrast, transformation with a temperature-sensitive mutant of RSV reversibly blocked the production of crystallins and lentoid bodies. These data show that given cellular genes can respond differently to distinct oncogenes. PMID- 3032461 TI - Induction of xanthophores from non-pigmented dermal cells of xanthic goldfish in vitro. AB - To identify precursor cells of xanthophores (xanthoblasts), non-pigmented cells without any phenotypic traits as pigment cells were isolated from the dermal tissue of xanthic goldfish with an adult color pattern and cultured in a medium containing 1 mM db-cAMP or 0.25 U/ml ACTH and 10% carp serum. These non-pigmented cells differentiated into xanthophores which showed a dendritic morphology and contained a large quantity of fluorescent pteridines and numerous vesicular inclusions. Sepiapterin was the major component, and the vesicles contained fuzzy material in addition to small membranous elements. The fluorescent pattern and the morphological characteristics indicated that the differentiated pigment cells were xanthophores of larval type. PMID- 3032463 TI - Regulation of human peripheral blood monocyte collagenase by prostaglandins and anti-inflammatory drugs. AB - Macrophages, which produce the collagenolytic enzyme collagenase, are commonly found at sites of connective tissue destruction in chronic inflammatory lesions. Since tissue macrophages are derived from circulating peripheral blood monocytes, we used these less-differentiated, more readily available cells to examine the production and regulation of collagenase. Human monocytes, isolated in large quantities by counterflow centrifugal elutriation, were shown to produce substantial amounts of collagenase when stimulated by concanavalin A (Con A) and to a lesser extent with lipopolysaccharide, while unstimulated monocyte cultures produced negligible collagenase. Collagenase was detected in the culture media within the first 24 hr of culture after activation with peak production at 48 hr. Analysis of the intracellular regulation of collagenase revealed that synthesis of this enzyme required a prostaglandin (PGE2)-dependent step since indomethacin inhibited enzyme synthesis was reversed by PGE2. Additionally, dibutyryladenosine cyclic monophosphate (dBcAMP) restored collagenase synthesis in indomethacin blocked cultures, indicating a PGE2-dependent generation of cAMP requirement for collagenase production similar to that demonstrated in experimental animals systems. In additional studies, anti-inflammatory drugs which are known to modulate connective tissue destruction were analyzed for their influence on monocyte-derived collagenase. Dexamethasone, colchicine or retinoic acid all inhibited collagenase synthesis by monocytes in a dose-dependent manner although the effect of these drugs on monocyte PGE2 synthesis differed. Dexamethasone inhibited PGE2 synthesis, which resulted in the suppression of collagenase. However, PGE2 production was unaffected by colchicine whereas retinoic acid caused a significant increase in PGE2 levels. Inhibition of collagenase synthesis by dexamethasone, but not colchicine or retinoic acid, could be reversed by PGE2 or phospholipase A2. These findings provide insight into the intracellular events regulating monocyte collagenase synthesis and also implicate monocytes as a target of anti-inflammatory agents which ameliorate connective tissue degradation associated with chronic inflammatory lesions. PMID- 3032464 TI - Study of DTH and resistance in Mycobacterium lepraemurium infection using a T cell line isolated from mice infected with Mycobacterium bovis (BCG). AB - A T-cell line of mixed phenotype (60% L3T4+, 40% Lyt-2+) was isolated from mice infected with Mycobacterium bovis (BCG). This line responded to M. lepraemurium and BCG but not to M. leprae and produced TCGF spontaneously. It also produced factors which stimulated macrophages to secrete hydrogen peroxide and superoxide anion. In vivo studies showed that only L3T4+ cells were required to transfer DTH responses and that Lyt-2+ cells suppressed this response. Both L3T4+ and Lyt-2+ cells were required to inhibit M. lepraemurium multiplication in vivo. PMID- 3032465 TI - Preexposure of resting B cells to interferon-gamma enhances their proliferative response to subsequent activation signals. AB - In this report we demonstrate that pretreatment of resting splenic B cells with IFN-gamma increases their mitogenic response to subsequent activating stimuli. This effect is completely blocked by neutralizing anti-IFN-gamma antibodies. By contrast, a similar effect induced by partially purified BCGF is not completely inhibited by anti-IFN-gamma antibody, inferring that as in the mouse, a B-cell specific factor may also induce increased responsiveness to mitogens in resting B cells. The mechanism of this response was analyzed. Phenotypic and cell cycle analyses of the IFN-gamma-treated cells following activation were not significantly different from control cells with respect to kinetics, although as expected from thymidine uptake, more cells were actively cycling. Even when a very early manifestation of cell activation, Ca2+ flux was examined, no response to IFN-gamma alone was evoked, and the response to subsequent activation was identical to that of control cells. These data show that IFN-gamma did not directly activate B cells, but primed B cells in a manner which amplified subsequent mitogenesis. PMID- 3032467 TI - Amplification of the c-myc gene and the elevation of its transcripts in human ovarian tumor lines. AB - c-myc gene was amplified in the human ovarian tumor line OS-4 5-6-fold and in the OvCa-1 line 3-4-fold. The relative amount of the transcript from the c-myc gene to the total RNA in OS-4 was 6-fold that for normal cells. PMID- 3032466 TI - Possible involvement of a Na+/H+ antiporter in the stimulation of hexose transport in Swiss 3T3 cells by a phorbol ester and growth factors. AB - Phorbol-12,13-dibutyrate, epidermal growth factor, and insulin raised the intracellular pH ([pH]i), presumably through the activation of a Na+/H+ antiporter. Addition of amiloride or replacement of extra-cellular Na+ by choline which abolishes the cytoplasmic alkalinization prevented the stimulation of hexose transport by these agents. Furthermore, monensin, a Na+/H+ ionophore which increases the [pH]i, stimulated hexose transport. This stimulation was also prevented by the replacement of extra-cellular Na+ by choline. These observations suggest that stimulation of the Na+/H+ antiporter may have stimulated the increase in hexose transport. PMID- 3032468 TI - [The proper time of spraying the leaves of Coix lacrymajobi with phosphorus]. PMID- 3032469 TI - Hepatocellular carcinoma presenting extrahepatic obstructive jaundice due to bile duct invasion--clinicopathological study of two cases. AB - Two rare cases of autopsy and surgery presenting extrahepatic biliary obstruction due to intrabile-duct growth of hepatocellular carcinoma were reported. Clinically obstructive jaundice was predominant in comparison with the other symptoms in both cases. In one autopsy case, hepatocellular carcinoma developed in the right lobe of the cirrhotic liver (posthepatitic). It involved the secondary branch of the right hepatic duct and grew into the common hepatic duct. In the other case of surgical operation, hepatocellular carcinoma, which developed in the posterior portion of the right lobe of the cirrhotic liver (posthepatitic), destroyed the posterior wall of the bifurcation of the bilateral hepatic duct and obstructed the common hepatic duct due to the intraductal cancer growth. From the site of the bile duct invasion or permeation by the tumor, two cases were classified into the peripheral (the former case) and proximal (the latter case) types, respectively. Furthermore, as far as obstructive jaundice is clinically concerned, the possibility should be kept in mind that hepatocellular carcinoma may proliferate into the large bile ducts, apart from that of cholangiocarcinoma or cholelithiasis. PMID- 3032470 TI - Extensive stage small cell carcinoma of the bronchus. A randomised study of etoposide given orally by one-day or five-day schedule together with intravenous adriamycin and cyclophosphamide. AB - Fifty-four patients whose disease had been staged as extensive small cell carcinoma of the bronchus were randomised to receive either CAV1 (cyclophosphamide 600 mg m-2 i.v., adriamycin 50 mg m-2 i.v., given on day 1, and etoposide 500 mg m-2 p.o. given on day 3) or CAV5 (cyclophosphamide and adriamycin given as for CAV1, etoposide 500 mg m-2 given in divided dose over days 3-7) on a 21-day schedule. The two regimens proved comparable (CR + PR 55% vs 56%), and the survival curves were virtually superimposable (median survival: CAV1, 8 months; CAV5, 9 months). Only five patients are still alive. The toxicity of the two treatments was similar. The scheduling of etoposide over 1 or 5 days seemed clinically unimportant in this study, perhaps because of concurrent use of other effective chemotherapy drugs. PMID- 3032471 TI - Pathophysiology of acute renal failure following living Escherichia coli injection in rats: high-energy metabolism and renal functions. AB - The energy metabolism of kidney and renal function were studied in rats following an IV injection of living Escherichia coli. Energy charge (ATP + 0.5 ADP/ATP + ADP + AMP) decreased throughout the period studied. Total and ouabain-sensitive Na-K ATPase activity of renal cortex homogenate decreased markedly at 3 hr followed by gradual recovery. Polyulia was seen at 3 and 6 hr followed by oliguria at 12 hr after E. coli injection. PSP excretion test showed a marked decrease throughout the time course. In contrast, creatinine clearance decreased only at 12 hr. From these results, it was clarified that the renal insufficiency following bacteremia occurs in two different stages; the early stage with a high urinary output accompanied by decreased Na-K ATPase activity suggesting deterioration of proximal tubular functions and the late stage with oliguria in which glomerular filtration is severely depressed. In both stages, renal energy metabolism is markedly disturbed. PMID- 3032472 TI - Propranolol treatment externalizes beta-adrenergic receptors in guinea pig myocardium and prevents further externalization by ischemia. AB - Purified sarcolemmal and light vesicle (intracellular) fractions of beta adrenergic receptors were used to examine the effects of propranolol on receptor translocation in guinea pig heart. Guinea pigs were given propranolol (0.15 mg/kg/hr) via minipumps for 7 days and either killed or made ischemic for 1 hour via a coronary ligature. Propranolol treatment led to an externalization of beta receptors from light vesicle to sarcolemmal fractions. This externalization increased the number of surface beta-adrenergic receptors that were functional, as assessed by isoproterenol-stimulated adenylate cyclase activity. After chronic propranolol treatment, ischemia did not further alter receptor distribution. These results suggest that externalization of beta-adrenergic receptors from a light vesicle fraction to the sarcolemma contributes to up-regulation of beta receptors that occur in response to both propranolol treatment and ischemia. Because propranolol-treated animals show blunting in externalization after myocardial ischemia, propranolol treatment and myocardial ischemia appear to access the same pool of intracellular beta-adrenergic receptors. Depletion of this pool of receptors along with receptor blockade may thus contribute to the mechanism by which the drug is efficacious in preventing some adverse effects of ischemia. PMID- 3032474 TI - Endothelium-dependent modulation of cGMP levels and intrinsic smooth muscle tone in isolated bovine intrapulmonary artery and vein. AB - The role of the endothelium in modulating cyclic nucleotide levels and intrinsic smooth muscle tone was studied in isolated rings of bovine intrapulmonary artery and vein. Cyclic 3',5'-guanosine monophosphate (cGMP) levels were threefold to fourfold higher in unrubbed artery and vein than in vessels that had been denuded of endothelium. Cyclic 3',5'-adenosine monophosphate (cAMP) levels were twofold higher in unrubbed than in endothelium-denuded artery, but no differences were observed in veins. Methylene blue, an inhibitor of guanylate cyclase, decreased cGMP but not cAMP levels, and this was accompanied by increases in smooth muscle tone. M&B 22,948, an inhibitor of cGMP-phosphodiesterase, increased cGMP but not cAMP levels, and this was accompanied by decreases in smooth muscle tone. Unrubbed vessels were more sensitive than endothelium-denuded vessels to the actions of both methylene blue and M&B 22,948, and this may be attributed to endothelium-dependent increases in cGMP turnover. Moreover, unrubbed vessels were more sensitive than endothelium-denuded vessels to contractile responses to phenylephrine and potassium, and these responses were potentiated by methylene blue and attenuated by M&B 22,948. Although indomethacin lowered cAMP levels in unrubbed artery, no changes in tone or contractile responsiveness were observed. A consistent observation was that the smaller branches of unrubbed but not endothelium-denuded intrapulmonary artery and vein had higher levels of cGMP but not cAMP, were sensitive to endothelium-dependent vasodilators, were more sensitive to methylene blue, and would not maintain a steady level of submaximal tone to phenylephrine when compared with larger branches from a common vascular bed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3032473 TI - Intracellular K+ activity, intracellular Na+ activity and maximum diastolic potential of canine subendocardial Purkinje cells from one-day-old infarcts. AB - The basis for the reduced maximum diastolic potential of canine cardiac subendocardial Purkinje fibers surviving one day after extensive transmural infarction was investigated, using double-barrel potassium and sodium ion sensitive microelectrodes. The maximum diastolic potential of Purkinje fibers in infarct preparations from the left ventricular apex measured during the first hour of superfusion in a tissue bath was -50.1 +/- 13.7 mV, a value markedly reduced from the value in control Purkinje fibers from noninfarcted preparations (-85.0 +/- 4.5 mV). The intracellular potassium ion activity was reduced by 50.4 mM during this time (intracellular potassium ion activity equals 61.6 +/- 16.1 mM, as compared to control intracellular potassium ion activity of 112 +/- 19.8 mM). The potassium equilibrium potential was reduced by 16.0 mV (from -97.2 +/- 4.7 mV in controls to -81.2 +/- 6.9 mV), thus accounting for about one half of the reduction in the maximum diastolic potential. After 6 hours of superfusion, the maximum diastolic potential increased to -78.9 +/- 8.7 mV (still significantly less than control). The potassium equilibrium potential had largely recovered (-93.8 +/- 5.9 mV). The intracellular sodium ion activity of Purkinje fibers in the infarcts (15.6 +/- 6.9 mM) was elevated during the first hour of superfusion by 6.2 mM compared to control (9.4 +/- 2.6 mM), and this was only 12% as much as the initial intracellular potassium ion activity decrease. Sodium ion activity after 3-6 hours of superfusion was not significantly different than normal (12.1 +/- 4.9 mM). In conclusion, only a portion of the maximum diastolic potential changes can be explained by a reduction of the potassium equilibrium potential. It is likely that change(s) in the cell membrane sodium-potassium pump's function and in the membrane conductance are also involved. Furthermore, the lack of a compensatory increase in intracellular sodium ion activity accompanying the large reduction of intracellular potassium ion activity may be a consequence of the cellular acidosis, which is known to occur during myocardial ischemia. PMID- 3032475 TI - Relation of arrhythmias and electrolyte abnormalities to survival in patients with severe chronic heart failure. AB - To investigate the determinants of mortality in patients with chronic congestive heart failure, we prospectively evaluated 84 patients with this disorder who underwent detailed biochemical, clinical, and functional tests at the time of initial evaluation and were then followed for 12 to 52 months (mean 31). During this period of follow-up, 58% of the patients died, of whom 71% died suddenly. The most important pretreatment predictor of mortality in these patients was the frequency of ventricular extrasystoles, followed by echocardiographic fractional shortening, a diagnosis of coronary artery disease, and duration of treadmill exercise. The finding of hypokalemia and hyponatremia in these patients at the time of entry into the study was associated with a poor prognosis by univariate analytical methods, but these electrolyte abnormalities did not provide independent prognostic information. The presence of ventricular arrhythmias was related to the severity of left ventricular dysfunction, exercise intolerance, and neurohormonal activation, suggesting that such arrhythmias are multifactorial in origin and may not simply be related to electrolyte abnormalities. PMID- 3032476 TI - The importance of defining left ventricular area at risk in vivo during acute myocardial infarction: an experimental evaluation with myocardial contrast two dimensional echocardiography. AB - Because the left ventricular "area at risk" is the most important determinant of ultimate infarct size, it would be useful to know the size of the area at risk during acute myocardial infarction to make therapeutic decisions. We therefore performed a series of experiments in four groups of dogs. In group I dogs (n = 15) we attempted to determine whether current methods of assessing left ventricular function during acute myocardial infarction reflect the true size of the area at risk. At each of two to five sequential stages, a more proximal coronary occlusion was performed to produce a larger area at risk until cardiovascular collapse occurred. At each stage, the area at risk (measured by myocardial contrast echocardiography), hemodynamic variables, and left ventricular ejection fraction (LVEF) were measured. Hemodynamic variables became abnormal when the area at risk was large (25% to 40% of the left ventricle), whereas LVEF became abnormal when the area at risk was of moderate size (18%). When cardiac output and LVEF were normalized to baseline values, a close inverse relationship was noted between these variables and area at risk. In contrast, there was a poor relationship between normalized mean arterial pressure and area at risk (r = .42). In group II dogs (n = 9) the area at risk was measured serially over 6 hr after coronary occlusion. The size of the area at risk remained unchanged regardless of the transmural extent of the ultimate infarct. The circumferential endocardial extent of the area at risk closely predicted the circumferential endocardial extent of the infarct at 6 hr in eight of nine dogs that developed an infarct. Group III dogs (n = 7) underwent the same protocol as group II dogs, but the duration of occlusion was 3 hr. The circumferential endocardial extent of the area at risk closely predicted the circumferential endocardial extent of the infarct. Group IV dogs (n = 5) underwent subtotal coronary occlusion. Although regional wall motion abnormalities were noted in this group, no area at risk could be defined. We conclude that although a close inverse relationship is noted between normalized cardiac output and area at risk, the absolute values for cardiac output and other hemodynamic variables become abnormal only when the area at risk is large (25% to 40%); measurement of LVEF may provide a better assessment of the size of the area at risk than hemodynamic variables.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3032477 TI - An unusual ultrastructural neutrophil abnormality of unknown function. PMID- 3032478 TI - Alterations of carboxypeptidases N activities in patients with thyroid dysfunction. AB - Serum carboxypeptidases N (EC. 3.4.17.3) were determined spectrophotometrically in 87 patients with disturbances of thyroid function and in 131 euthyroid individuals, including 33 women taking estrogens for contraception. Carboxypeptidases N (CN) can be subdivided into CN1 and CN2, according to variable substrate affinity. In addition, measurements of blood pressure and in vitro tests of thyroid function were performed. In euthyroid controls, CN1 was negatively correlated with age. No significant differences between CN1 and CN2 have been observed with regard to sex. In patients with hyperthyroidism, the mean values of both enzymes were elevated, but this tendency proved to be significant only for CN2. In hypothyroid patients CN1 and CN2 levels were normal. Elevations of CN1 and CN2 in the hyperthyroid state seem not to be related to underlying immunological processes but to the thyroid hormone excess itself. In euthyroid women taking estrogens for contraception CN2 was also elevated. PMID- 3032479 TI - Is paraoxon hydrolytic activity in serum predictive of myocardial infarction? PMID- 3032480 TI - Mycoplasma, chlamydia and viruses in the female genital tract and infertility: clinical experience. AB - In the University of Padua during the last ten years the chlamydia trachomatis, mycoplasma and viruses were studied in human infertility. The results show the influence on chlamydia and ureaplasma urealyticum, in some cases with different pathogenetic mechanisms. The results are discussed. PMID- 3032481 TI - Dose response relationship of relaxin on the pubic ligament of the mouse. AB - In the present work, the dose-response relationship of highly purified porcine relaxin has been examined on broadening of the pubic ligament in mice. Using the method of Steinetz et al. with 7 days of oestriol priming, higher sensitivity of the pubic ligament was attained in mice with an original weight of 10 g than in those with a weight of 20 g. S-shaped curves were obtained; by increasing the relaxin doses after maximal effect had been reached a decreased action was observed. With presomen priming, a relaxin effect was established after only 2 days instead of the usual 8 days but the action was markedly less than in the Steinetz test. PMID- 3032482 TI - The relative extent of glycation of haemoglobin and albumin. AB - The level of non-enzymatic glycation of a protein is thought to depend on the number of sites available for reaction, the half-life of the protein and the ambient concentration of glucose. Accordingly, the modification of two blood proteins with a similar number of potential sites but different survival times was examined in non-diabetic patients by periodate oxidation and by reduction with [3H]borohydride. The amount of glycation of haemoglobin and its sub fractions HbA1 and HbA1c were determined to be 0.44, 2.42 and 2.24 mol/mol respectively and the corresponding value for albumin was 0.37 mol/mol protein. Amino acid analysis showed that the epsilon amino groups of albumin were more extensively modified than they were in haemoglobin and thus it is concluded that the average rate of reaction of the lysine residues in albumin is markedly faster than in haemoglobin. PMID- 3032483 TI - Peripheral neuropathy with gammopathy responding to plasmapheresis. AB - Two patients with subacute sensorimotor peripheral neuropathy were found to have benign gammopathies. They both responded to treatment by plasmapheresis. We suggest that a vigorous search for a paraprotein should be made in all undiagnosed patients presenting with peripheral neuropathy, including electrophoresis with isoelectric focusing. Plasmapheresis may be a treatment modality that should be explored in such cases. PMID- 3032484 TI - IgG subclass potential of surface IgM-negative and surface IgM-positive human peripheral blood B cells. AB - To examine the IgG subclass potential of B cells at various stages of maturation, human peripheral blood B cells were separated into surface IgM-positive (sIgM+) and surface IgM-negative (sIgM-) cells by panning techniques and cultured with pokeweed mitogen, LPS, or Epstein-Barr virus. At the end of 7 days, cells were harvested, counted, spun onto slides, fixed, and stained with subclass-specific monoclonal antibodies. In all experiments with all mitogens, there was an enrichment of IgG2 plasma cells in sIgM+ cultures compared to the sIgM- cultures (P less than 0.0001). In the Epstein-Barr virus-stimulated cultures there was also a statistically significant enrichment for IgG3 in the sIgM+ cultures (P less than 0.01). In contrast, sIgM- cultures were always enriched for IgG1 plasma cells (P less than 0.0001). In the pokeweed mitogen-stimulated cultures, the sIgM cultures also contained a higher proportion of IgG4 plasma cells than did the sIgM+ cultures (P less than 0.01). These results demonstrate that the IgG subclass potential of immature, surface IgM-positive precursors of IgG plasma cells differs from that of more mature surface IgM-negative precursors. PMID- 3032485 TI - The role of colony stimulating factors in leukaemogenesis. PMID- 3032486 TI - Lafora disease: a quantitative morphological and biochemical study of the cerebral cortex. AB - Morphological and biochemical studies were performed on a brain biopsy from a patient with typical Lafora disease. Qualitative morphological investigation of the cortex showed that the Lafora bodies were most abundant in layers III and V of the cerebral cortex. They were exclusively located in the neurons and their processes. Quantitative morphological investigation of the cerebral cortex revealed abnormalities of the pyramidal cells of layers III and V. The neurons were often slightly atrophic. There was a reduction in the number of dendrites, in maximal dendritic length, and in the number of spines on the apical dendrites and on the side branches. Because of the biochemical study of the pyruvate metabolism of the gray and white matter of the cortex did not show abnormalities, a mitochondrial dysfunction is not likely. PMID- 3032487 TI - Mitochondrial paracrystalline inclusions in the peroneus brevis muscle of patients with peripheral neuropathy. AB - In a series of 50 consecutive biopsies of peroneus brevis muscle (PBm) taken from patients with ascertained or suspected polyneuropathy in the course of sural nerve biopsy, we found a high incidence (26%) of intramitochondrial paracrystalline inclusions (MPI). Five out of these 13 patients were also submitted to an additional biopsy of a proximal muscle, which in no case confirmed the finding of MPI. Six out of the 13 patients with MPI were affected by diseases with a presumably important ischemic component. The mean age of patients with MPI was quite elevated (60.7), and the difference in distribution of age between patients with and without MPI was statistically significant. No significant difference in histochemical changes was found between the group of MPI patients and a control group of age-matched patients without MPI, thus excluding that MPI in the PBm are specifically associated with other neurogenic or myopathic aspects. We conclude that aging and, probably, ischemia are largely responsible for the frequent presence of MPI in the PBm. In addition, factors intrinsic to the muscle itself, possibly related to morphological, physiological, or biochemical peculiarities, may also influence the development of MPI. PMID- 3032489 TI - [Scanning electron microscopy and X-ray spectrometric studies of Hirano bodies: aluminum accumulation in Hirano bodies]. PMID- 3032488 TI - [Metronidazole neuropathy]. PMID- 3032490 TI - Variability of central conduction in the course of multiple sclerosis. Serial recordings of evoked potentials in the evaluation of therapy. AB - In 27 normal subjects evoked potentials (EPs) were serially measured across an interval of 3.5 months. The variation in latencies of the P100 component of the VEP and of the components N13 and N20 of the SEP did not exceed 12% of the absolute values of these latencies on the first recording. The VEP and SEP were also serially recorded over the same interval in 35 patients with 'clinically definite' multiple sclerosis. Changes of the latencies exceeding those of normal subjects were noted in 30% of the cases. The changes in the EPs showed no correlation with the variations in the overall clinical disability. It is argued that EPs are probably more sensitive to changes in function of parts of the white matter than the clinical examination and are useful in the quantification of these changes. As a model for future studies, EPs were used in the 'evaluation' of ACTH therapy. It appeared, that therapy did not have effect on the EPs. PMID- 3032491 TI - Inhibition of renal clearance of furosemide by pentopril, an angiotensin converting enzyme inhibitor. AB - The pharmacokinetic interaction between pentopril (250 mg) and furosemide (40 mg) was studied in 12 normal healthy volunteers after oral administration of each drug alone and in combination. No significant changes in any pharmacokinetic parameters of pentopril or its active metabolite (CGS 13934) were observed on coadministration of furosemide. In contrast, pentopril induced significant changes in disposition of furosemide. Pentopril decreased renal clearance (CLR) of furosemide by 54% and the fraction excreted unchanged in urine also decreased by 55%. However, such decrease in CLR of furosemide was compensated by a simultaneous increase in glucuronidation (by 200%), resulting in a slight increase in systemic clearance (decreased AUC). Systemic bioavailability of furosemide appears to be unchanged in the presence of pentopril (0.46 vs. 0.41). No effect of pentopril on plasma protein binding of furosemide was detected. In spite of the decreased CLR and urinary excretion rate of furosemide, the urinary output (1749 vs. 1774 ml/6 hr) and Na+ excretion (757 vs. 816 mEq/6 hr) remained almost unchanged. These findings suggest that total furosemide (unchanged and glucuronide) might contribute to diuresis and natriuresis rather than the unchanged furosemide alone. Because of unchanged pharmacodynamic effect, such pharmacokinetic interaction may not require any dosage adjustment for furosemide on pentopril coadministration. PMID- 3032492 TI - The effect of a dentifrice containing soluble pyrophosphate and sodium fluoride on calculus deposits. A 6-month clinical study. PMID- 3032493 TI - Peripheral and autonomic nerve function in long-term insulin-dependent diabetes. AB - In a cross sectional study, motor nerve conduction velocity (MNCV) and sensory nerve conduction velocity (SNCV), beat-to-beat variation (BBV) at rest and speed of pupillary dilatation (SPD) have been investigated in 127 nonketonuric long term insulin-dependent diabetics aged 19-72 yr and in age-matched control subjects. 84% of the patients had electrophysiologic abnormalities, 58% had symptomatic peripheral neuropathy, 35% had abnormal cardiac parasympathetic tests and 26% had abnormal pupillary tests. The most frequent pathologic feature was a decreased sural SNCV (66%). Among the patients without symptomatic peripheral neuropathy, 71% showed electrophysiologic abnormalities. MNCV and SNCV in median, peroneal and sural nerves correlated with BBV (p less than 0.005), but only peroneal MNCV was related to SPD (p less than 0.005). There was also a relationship between BBV and SPD (p less than 0.05). Glycosylated hemoglobin (HbA1) levels correlated inversely with median MNCV (p less than 0.001), and SNCV (p less than 0.03), peroneal MNCV (p less than 0.05) and sural SNCV (p less than 0.05). Patients with abnormal peripheral or autonomic nerve function tests or symptomatic peripheral neuropathy had significantly higher HbA1 levels than those with normal tests (p less than 0.04) or asymptomatic patients (p less than 0.01). Median MNCV and SNCV, sural SNCV and BBV deteriorated with age (p less than 0.05) and median SNCV and peroneal MNCV deteriorated with the duration of diabetes (p less than 0.001). Our findings show an association between peripheral and autonomic nerve dysfunction in long-term insulin-dependent diabetics.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3032494 TI - [Phantom substances for quantitative evaluation of MRT images. III. Effect of various protein concentrations on MRT intensity values]. AB - The use of heavy water (D2O) in paramagnetic agar phantoms can yield tissue equivalent values of the proton spin density p. In this manner all the MRI parameters that are typical of tissues can be realised in phantom substances. It is shown that the use of heavy water results in some enhancement of relaxation time, T2, that can be easily controlled via the agar concentration. PMID- 3032496 TI - Use of the modified Cope introduction set for transhepatic removal of obstructed Carey-Coons biliary endoprosthesis. AB - A new method of replacing an occluded biliary endoprosthesis by using the modified version of the Cope catheter introduction set is described. PMID- 3032495 TI - [Sonographic diagnosis of liver tumors. Results of comparative studies of ultrasound, computerized tomography, laparoscopy, biopsy and scintigraphy in 413 patients]. AB - Based on a retrospective study of 413 patients, the accuracy of sonography compared to CT, laparoscopy, biopsy and scintigraphy in the primary diagnosis of liver masses is shown. False positive sonographic reports are analysed in retrospect. 190 of 338 solid space-occupying growths of the liver have sonographically been called definite growths and 148 have been considered suspicious of being such growths. 49 of the cystic lesions were considered definitely cystic and 26 suspicious. Sonography made correct positive diagnoses in 91.8% of the cysts and in 88.9% of the solid growths. The diagnostic accuracy of solid growths (without further specification) was 90%. Among the sonographically suspicious ones the specific diagnosis was correct in 80.8% of the cystic lesions and in 44.5% of the solid lesions and a correct diagnosis without further specification was made in 55.4%. The majority of false positive diagnoses corresponded to normal findings in other methods. The number of false positive diagnoses in sonography in contrast to CT depends to a high degree on the experience of the physician. Before the use of other more invasive methods the sonographically suspicious findings should therefore be double-checked by an experienced colleague with several years of sonographic experience. PMID- 3032497 TI - The Carey-Coons percutaneous biliary endoprosthesis: a three-centre experience in 87 patients. AB - Eighty-seven patients with malignant obstruction of the biliary tract from three centres and deemed unsuitable for surgery underwent insertion of the 'Carey Coons' transhepatic endoprosthesis. It was successfully introduced in all, with early relief of cholestasis in 97%. The 30 day mortality rate was 11.5%, which is lower than reported in series using different endoprostheses. Two fifths of patients had complications, especially cholangitis. Despite its ease of insertion the design of the endoprosthesis may be related to the high incidence of cholangitis and to formation of biliary sludge and occlusion found in follow up. Although the anchoring threads prevented migration, they may have played a role in infection at the skin entry site in four patients and in tumour seeding to the skin in a further two. Still further improvements in endoprosthesis design are desirable. PMID- 3032498 TI - Second malignant neoplasms after childhood cancer: a report of three cases of osteogenic sarcoma. AB - The rising incidence of second malignant neoplasms after childhood cancer, whilst due in part to increasing numbers of survivors, is also thought to be related to increasingly more intensive combined modality treatment schedules. Three illustrative cases are reported in which radiation therapy in childhood for the first cancer is thought to have been a significant aetiological factor in the pathogenesis of the second malignancy (which in all three patients was an osteogenic sarcoma). PMID- 3032499 TI - Aldose reductase inhibition, glomerular metabolism, and diabetic nephropathy. PMID- 3032500 TI - The polyol pathway in dysfunction of diabetic peripheral nerve. PMID- 3032501 TI - New indices for selection of carbohydrate foods in the diabetic diet: hopes and limitations. PMID- 3032502 TI - Comparison of electron microscopy with three commercial tests for the detection of rotavirus in animal feces. AB - Three commercial test kits were evaluated to detect the presence of rotavirus antigens in bovine, porcine, and turkey feces. Two of the assays, Rotalex (Medical Technology Corporation, Somerset, NJ) and Virogen-Rotatest (Wampole Labs, Cranbury, NJ) are latex agglutination tests (LA), while the third, Pathfinder (Kallestad, Austin, TX) is an enzyme immunoassay. The clinical usefulness of these assays was elevated by comparing their results with those of direct electron microscopy (EM). A total of 135, 92, and 211 samples of animal feces were tested by Rotalex, Virogen, and Pathfinder, respectively. All samples were examined by EM as a reference procedure. The overall agreement of the three commercial assays with EM was 53%, 66%, and 83% for Rotalex, Virogen, and Pathfinder, respectively. Based on these results, we consider Pathfinder as an attractive alternative to EM for the detection of rotavirus in animals. Of the two LA tests, Virogen was found to be a little more sensitive and specific. PMID- 3032503 TI - In vitro susceptibility of cytomegalovirus isolates from immunocompromised patients to acyclovir and ganciclovir. AB - Fifty-four isolates of cytomegalovirus (CMV) from 25 immunocompromised patients with CMV infections were examined in a plaque reduction assay to determine in vitro susceptibilities to both acyclovir and ganciclovir. Isolates were approximately 25-fold more sensitive to ganciclovir than to acyclovir. The mean +/- SD ID50 for all isolates to acyclovir was 63.1 +/- 30.2 microM, (median, 52.3 microM; range, 16.7-146.4 microM). The mean ID50 for all isolates to ganciclovir was 2.50 +/- 1.27 microM, (median, 2.15 microM; range, 0.65-7.11 microM). Exposure to acyclovir or ganciclovir for periods of 2-5 wk did not alter the mean susceptibility of clinical isolates. However, one CMV strain isolated from a renal transplant patient after 14 days of acyclovir therapy displayed the lowest susceptibility of all strains tested (acyclovir 146.4 microM; ganciclovir 7.11 microM) and may represent selection of a resistant virus population. PMID- 3032504 TI - New drug treatment of hypertension. PMID- 3032505 TI - Geriatric malnutrition: recognition and prevention. PMID- 3032506 TI - Advances in the diagnosis and management of congestive heart failure. PMID- 3032507 TI - Lectin inhibition and kinetics of microsomal K+-dependent p-nitrophenyl phosphatase of frog epidermis. AB - The specific activity of K+-dependent p-NPPase (paranitrophenylphosphatase) from frog (Rana ridibunda) epidermis microsomal preparation was determined. The activity was proportional to time of incubation and protein concentrations under our assays conditions. Optimal phosphatase activity was at pH from 8 to 9 and over 35 degrees C. 10(-3) M ouabain inhibited 100% of the activity and the Ki was estimated about 5 X 10(-5) M. The Km for p-NPP was 3.8 mM and 2.1 for K+. The lectins GSI and GSII produced 80-90% of non-competitive inhibition of the activity. 50% of inhibition by GSI was obtained at 2 micrograms/ml. The Km for p NPP did not change but the Vmax of activity was clearly reduced for both GSI and GSII lectins. PMID- 3032508 TI - An ion-exchange mechanism of cartilage calcification. AB - The provisional calcification of epiphyseal cartilage involves deposition of hydroxyapatite (calcium phosphate) crystals in an extracellular matrix consisting principally of Type II collagen and cartilage proteoglycan. A mechanism is now proposed to explain how epiphyseal cartilage calcification is initiated. Calcium exists at high concentration in cartilage, but is mainly bound to the anionic groups of proteoglycans, and thus is unavailable for precipitation. A local increase in phosphate concentration displaces calcium ions from proteoglycan by an ion-exchange effect, raising the Ca X PO4 product above the threshold for precipitation of hydroxyapatite. Evidence for this hypothesis has been derived from studies of the effect of phosphate on the binding of calcium to cartilage proteoglycan, and on hydroxyapatite formation in the presence of chondroitin sulfate. PMID- 3032509 TI - Dynamics of collagen accumulation and activity of collagen-degrading enzymes in the liver of rats with carbon tetrachloride-induced hepatic fibrosis. AB - Liver fibrosis in rats was induced by repeated subcutaneous injections of carbon tetrachloride. Total collagen, soluble and insoluble collagen fractions as well as type I and type III collagen content in the liver were subsequently measured over a 3-18 week period. Liver collagen was found to increase exponentially during this time. Insoluble collagen accumulated more rapidly than soluble forms, and the accumulation of type III collagen was relatively greater than type I collagen. Changes in specific liver enzymes were also observed. Collagenase, collagenolytic cathepsin and collagen peptidase activities all increased. Levels of collagen-degrading enzymes increased rapidly during the first weeks of fibrosis-induction, and were followed by a more gradual increase during the remainder of the study. PMID- 3032510 TI - Liver disease in renal transplant patients treated with azathioprine or ciclosporin. PMID- 3032511 TI - Experimental uremic cardiomyopathy--fact or fiction? PMID- 3032512 TI - Hypertension in diabetes mellitus. PMID- 3032513 TI - Segmental atresia of the transverse colon in a foal with concurrent equine herpes virus-1 infection. AB - Segmental atresia of the transverse colon was observed at necropsy in a neonatal foal. The dorsal and ventral components of the large colon were fused, and ended blindly. The small colon was collapsed and completely closed at its cranial end. The right and left dorsal and ventral colons were fused into one blind-ended tube. Histologically, eosinophilic intranuclear inclusion bodies demonstrative of Equine Rhinopneumonitis were present in the thymus. PMID- 3032514 TI - Unilateral neglect and constructional apraxia in a right-handed artist with a left posterior lesion. AB - A rapidly growing tumor in the left posterior parietal lobe of a right-handed, 72 year old artist resulted in bilateral neglect and constructional apraxia that were greater on the right side of his painting than on the left. These changes indicate that a left posterior parietal lesion alters visuospatial perception and constructional ability on both sides of a painting with the contralateral hemispace more impaired than the ipsilateral hemispace. Primitivization of emotional expressions on human faces also occurred. PMID- 3032515 TI - Inhibition of angiotensin-converting enzyme by perindopril diacid in canine oleic acid pulmonary edema. AB - To test the hypothesis that angiotensin II could be a mediator of acute lung injury, we studied the effects of perindopril diacid, a new angiotensin converting enzyme inhibitor, on hemodynamics, blood gases, lung mechanics, and extravascular lung water (EVLW). Twenty-four dogs were anesthetized, paralyzed and ventilated with a fraction of inspired oxygen of 0.4 in which pulmonary edema was induced by 0.1 ml/kg iv oleic acid. Perindopril diacid (1 mg/kg) was administered iv either before (eight dogs) or 100 min after (eight dogs) oleic acid injection. In the control group (eight dogs) not treated with perindopril diacid, 150 min after oleic acid injection, PaO2 changed from 193 +/- 7 (mean +/- SEM) to 55 +/- 4 torr, venous admixture from 3 +/- 1% to 52 +/- 5%, cardiac index from 4.1 +/- 0.3 to 3.1 +/- 0.3 L/min X m2, mean pulmonary artery pressure from 13 +/- 1 to 17 +/- 1 mm Hg, dynamic compliance from 90 +/- 8 to 46 +/- 7 ml/cm H2O, and EVLW from 165 +/- 25 to 750 +/- 92 ml/m2. Administration of perindopril diacid reduced systemic BP by 20% but did not affect other hemodynamic variables, blood gases, or dynamic compliance. Maximum increases in EVLW were from 169 +/- 16 to 615 +/- 54 ml/m2 in the pretreated group and from 188 +/- 23 to 675 +/- 56 ml/m2 in the treated group (no significant difference from the control group). However, pretreatment with perindopril diacid significantly (p less than .05) slowed the rise in EVLW, which was lower 60 and 90 min after oleic acid injection compared to untreated dogs. Plasma renin activity and angiotensin I concentration increased after oleic acid injection.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3032516 TI - Selenite cataract: a review. AB - Selenite cataract is a fairly recently described, experimental animal model for cataract (1). Selenite cataract has been extensively characterized histologically (2) and biochemically (3,4). The model has been particularly useful for studies on the roles of calcium accumulation and lens proteolysis in cataract formation (4). This review describes current knowledge of the biochemical mechanism for selenite cataract and indicates how the model may be used for further understanding of cataractogenesis in general. PMID- 3032517 TI - The effects of forskolin on cyclic AMP, intraocular pressure and aqueous humor formation in rabbits. AB - Forskolin was used to study cyclic AMP-mediated regulation of aqueous humor dynamics in rabbits. Crystalline forskolin was solubilized in oil and its pharmacological effects were studied both in vitro and following topical ocular administration. In vitro, using cultured corneal epithelial cells, forskolin rapidly stimulated cyclic AMP production and in vivo increased cyclic AMP concentration in the aqueous humor 10-fold following topical administration. The effect of topical forskolin on intraocular pressure and aqueous humor formation was determined in vivo using pneumatonometry and fluorophotometry, respectively. Forskolin caused a prolonged reduction of intraocular pressure and decreased aqueous humor formation. The ability of forskolin to potentiate the ocular hypotensive effect of epinephrine was investigated. Forskolin in combination with epinephrine caused a decrease in intraocular pressure of longer duration than either 0.1% epinephrine or 1% forskolin administered separately. Forskolin caused a small but significant increase in the permeability of the blood-aqueous barrier at the time of maximal intraocular pressure reduction. This effect on the blood aqueous barrier may explain the inhibitory effect of forskolin on aqueous humor formation. PMID- 3032518 TI - Immunoelectron microscopic localization of photoreceptor-specific markers in the monkey retina. AB - Antibodies for several molecules that function in the visual process were used to localize these molecules in primate rod and cone cells. These antibodies (monoclonal or polyclonal) were prepared against Interphotoreceptor Retinoid binding Protein (IRBP), S-antigen (S-Ag), opsin, alpha-transducin and also against cyclic GMP (cGMP). Lowicryl-embedded tissues were labeled with secondary antibodies linked to colloidal gold. Although IRBP is predominantly an extracellular protein, the relatively small amount found intracellularly was localized mainly in rods, with little in cones. Opsin, S-Ag and cGMP were found mainly in rod cell outer segments. A polyclonal antiserum raised against transducin-alpha purified from rod outer segments predominantly labeled rod cells, but an antiserum against the carboxyterminal decapeptide of transducin alpha labeled both rod and cone cells. Thus, most of these specialized molecules are present predominantly in rod cells, confirming major differences in components of the visual cycle in rods and cones. PMID- 3032520 TI - The management of malignant strictures of the bile duct. PMID- 3032519 TI - Corneal deturgescence during organ culture. AB - A simple method is described to monitor rabbit and human corneal deturgescence while the tissue is suspended in tissue culture media. The composition of the media is such that the cornea slowly thickens at 4 degrees but exhibits the classic "temperature reversal" thinning phenomenon when incubated at 23 degrees 37 degrees. Each cornea is cultured in a closed eye bank corneal viewing chamber during swelling and deswelling phases of an experiment, minimizing direct handling of the tissue. Optimal conditions for observing corneal deturgescence are described. This protocol has several advantages over corneal perfusion techniques and could be further developed for routine use in eye banks as a functional test of donor suitability for keratoplasty. PMID- 3032521 TI - Re-evaluation of the sublocalization of esterase D and its relation to the retinoblastoma locus by in situ hybridization. AB - In situ hybridization of a cDNA probe for the esterase D gene (ESD) was carried out on human chromosomes. The probe hybridized most strongly to 13q14.2 and 13q14.3. This observation raises doubts concerning the most recently published assignment of ESD to 13q14.1. A deletion in an individual with retinoblastoma was reported to separate the closely linked ESD and retinoblastoma (RB1) loci, placing ESD proximal to RB1. Quantitative in situ hybridization studies of this deletion do not confirm this interpretation. Rather, they suggest that ESD is missing from the deleted chromosome 13 and duplicated on the normal homolog. From these findings, we conclude that the deletion in this individual cannot be used to determine the orientation nor the sublocalization of ESD and RB1 within the 13q14 region. PMID- 3032522 TI - Lymphocyte subsets in lung cancer. AB - Altered cellular immune function has been demonstrated in patients with lung cancer, including decreased numbers of circulating lymphocytes and changes in the percentage of lymphocytes in various functional subsets. We quantitated lymphocyte subsets in 54 patients with lung cancer including patients with limited (stages 1 and 2) nonsmall cell lung cancer (NSCLC, n = 23), advanced (stage 3) NSCLC (n = 16), and small cell cancer (SCLC, n = 15). Serum albumin was decreased in 15 lung cancer patients, and lymphocyte subsets were separately evaluated in these patients. Lymphocyte populations in cancer patients were compared to those of nonsmokers and a smoking patient population. No difference from smokers was noted in patients with limited NSCLC. Patients with SCLC and advanced NSCLC had significantly decreased numbers of T-helper and T-suppressor cells (p less than 0.05). Patients with lung cancer and hypoalbuminemia had the greatest decrease in number of circulating T-helper cells (p less than 0.001). B lymphocytes were also decreased in patients with advanced NSCLC and patients with hypoalbuminemia (p less than 0.05). A decrease in population of T-lymphocytes subsets is frequent in patients with SCLC, advanced NSCLC, and lung cancer patients with hypoalbuminemia. PMID- 3032523 TI - Potentially deleterious effects of long-term vasodilator therapy in patients with heart failure. PMID- 3032524 TI - Exacerbation of pulmonary lymphangioleiomyomatosis by exogenous estrogens. AB - A 48-year-old woman with profound, rapidly progressive dyspnea requested a second opinion regarding the diagnosis and management of an undiagnosed interstitial process. One year prior to this evaluation, she had been placed on therapy with exogenous estrogens for the treatment of osteoporosis. During this therapy, she had a marked deterioration of her pulmonary status. Review of her open lung biopsy, which was obtained five years previously, revealed lymphangioleiomyomatosis. Discontinuation of estrogen therapy and treatment with tamoxifen were successful in stopping the progressive course. This patient's clinical course suggested an association between estrogen therapy and clinical deterioration during the menopause. PMID- 3032525 TI - Autonomic control of airway function in asthma. PMID- 3032526 TI - Effect of vidarabine in dimethyl sulfoxide vehicle on type 1 herpesvirus-induced cutaneous lesions in laboratory animals. AB - Vidarabine (9-beta-D-arabinofuranosyladenine) prepared in a 70% dimethyl sulfoxide vehicle was applied topically to type 1 herpesvirus-induced cutaneous lesions on guinea pigs and athymic nude mice. Treatments were 3 or 5 times daily for 7 days beginning 24 h after virus exposure. Against infections in guinea pigs induced by a thymidine kinase-positive virus strain, either treatment schedule effectively inhibited mean lesion score, lesion size, appearance of new lesions, and reduced lesion virus titers. Therapy was similarly effective against infections in guinea pigs induced by a thymidine kinase-negative virus strain, except that lesion virus titers were somewhat increased in animals treated 3 times daily. Treatment 5 times daily was most efficacious against both virus strains. Treatment 3 times daily of mice infected with a thymidine kinase negative virus was not effective, but treatment 5 times daily significantly inhibited lesion score and size and reduced lesion virus titer by 37%. Toxicity controls exhibited no signs of skin irritation, although guinea pigs treated 5 times daily experienced some transient weight loss. PMID- 3032527 TI - Inhibitory effect of Azone (1-dodecylazacycloheptan-2-one) on herpes simplex viruses. In vivo and in vitro studies. AB - Enhanced activity of antiviral agents, when mixed with a penetration enhancer like Azone (1-dodecylazacycloheptan-2-one) in topical preparations, is well recognized. These studies were undertaken to investigate whether Azone, per se, has any activity against herpes simplex (HSV) type 1 and type 2 viruses. When HSV 1 and HSV-2 were mixed with 5, 10 and 20% Azone, there was a 1-4.5 log10 decrease in virus titers. The efficacy of 5 and 10% Azone applied topically was evaluated during the treatment of HSV-1 cutaneous infections in guinea pigs. HSV-1 infection was produced by intradermally inoculating guinea pigs with 10(7.7) HSV 1/ml. The mean number of lesions per site was reduced by more than 50% in Azone treated sites as compared to no treatment (control) sites (p less than 0.01). Similarly, the surface area of each lesion in the Azone-treated sites was only 40% of that seen in control sites (p less than 0.001). More than 80% of the lesions on Azone-treated sites healed by day 4 as compared to only 3% on control sites (p less than 0.001). These data indicate that Azone, a lipophilic chemical, may have antiviral properties when used topically. PMID- 3032528 TI - [Herpes virus. Herpes infections and therapeutic implications]. PMID- 3032529 TI - [Clinical features and management of fulminating and silent pituitary apoplexy in ACTH-secreting tumors]. PMID- 3032530 TI - [Peripheral neuropathy associated with rheumatoid arthritis. Report of clinical and pathological findings of a sural nerve biopsy in 6 cases]. PMID- 3032531 TI - [Pericardial metastases in lung cancer. A report of 10 cases]. PMID- 3032532 TI - [Small cell carcinoma of the lung associated with syndrome of inappropriate secretion of antidiuretic hormone: report of a case and review of the literature]. PMID- 3032533 TI - Alcoholism: scientific basis and chemotherapeutic control. PMID- 3032534 TI - Turcot syndrome. Autosomal dominant or recessive transmission? AB - The authors analyzed a family in which three descendants presented with adenocarcinoma of the colon. In two of them the presence of colonic adenomatosis was observed. Another family member, a 13-year-old girl, presented with Turcot syndrome, that is, brain tumor associated with colonic adenomatosis. The nature of the hereditary transmission of Turcot syndrome is hence analyzed, discussing whether it happens through an autosomal recessive or a dominant gene. Undoubtedly the family has colonic adenomatosis, a disease considered of autosomal dominant transmission. Based on the clinical observation, the authors suggest that Turcot syndrome may be determined by an autosomal gene with a pleiotropic effect and variable expressivity. PMID- 3032535 TI - Upper gastrointestinal neoplasia in familial polyposis. AB - Upper gastrointestinal (UGI) endoscopy was performed in 41 asymptomatic American patients with familial polyposis to assess the prevalence of gastric and duodenal polyps and to characterize their pathological features. Eighteen patients (44%) had UGI endoscopic abnormalities. Six patients had both gastric and duodenal lesions. Eight patients had only gastric polyps, and four had duodenal polyps only. The presence of other extracolonic expressions of polyposis had a suggestive but statistically insignificant correlation with UGI polyps. Patients with familial polyposis and duodenal adenomatous polyps are at high risk for the development of periampullary cancer; screening and identification of these individuals is recommended. PMID- 3032536 TI - Nutritional recommendations and principles for individuals with diabetes mellitus: 1986. American Diabetes Association. PMID- 3032537 TI - No glycemic benefit from guar administration in NIDDM. AB - A randomized crossover study of 5-g guar minitablets against placebo, given three times per day with main meals for 8 wk, was done in 29 non-insulin-dependent diabetes mellitus (NIDDM) patients who had near-normal fasting plasma glucose concentrations on treatment with diet alone, additional sulfonylurea, or ultralente insulin. Guar did not reduce the excessive postprandial glycemic excursion, glycosylated hemoglobin values, basal plasma glucose concentrations, basal or incremental plasma C-peptide values, or body weight. There were few side effects with either guar or placebo therapy. Mean low-density lipoprotein cholesterol levels were significantly reduced (P less than .001) by guar administration (116 +/- 23 vs. 104 +/- 19 mg/dl). Guar additives did not improve the excessive postprandial glycemia found in NIDDM patients in whom near-normal fasting plasma glucose levels had been obtained. PMID- 3032538 TI - Fine-needle aspiration cytology in pancreatic endocrine tumors. AB - Pancreatic endocrine tumors (PETs) are relatively uncommon neoplasms. Although their histologic patterns have been widely studied, their cytologic features as they appear in fine-needle aspiration (FNA) specimens have rarely been reported. In this study, aspirates of seven PETs, four primary and three metastatic lesions (two to liver and one to bone), are described. The tumors occurred in seven men ranging in age from 37 to 72 yr. Six tumors presented as nonfunctioning masses and one produced Zollinger-Ellison syndrome. Three were located in the head of the pancreas and four in the body and tail. The pancreatic and liver aspirations were performed under computed tomographic guidance and the bone lesion, under fluoroscopy. The aspiration specimens were hypercellular. The tumor cells occurred singly and in small clusters. In three cases, there was a tendency toward acinar formations. In two cases, there were prominent, thin-walled, branching blood vessels with tumor cells attached to the vascular walls. The cells were round or polygonal with a moderate amount of finely granular, well defined cytoplasm. The nuclei were eccentrically located and round-to-oval--with one or two small nucleoli and finely granular, evenly-dispersed chromatin. The diagnosis was confirmed by immunocytochemistry (two cases) and electron microscopy (four cases) of the aspirated material and histology sections of the resected tumors (two cases). The results of this study demonstrate that FNA is a useful method to establish the diagnosis of PETs. PMID- 3032539 TI - Neuron-specific enolase-positive rosettes in nephroblastoma: a possible diagnostic pitfall in aspiration cytology. AB - Neuron-specific enolase-positive cells, some arranged in rosettes, were identified in smears obtained in fine-needle aspiration of a left retroperitoneal tumor found in a 3-yr-old boy. Nephrectomy showed a typical nephroblastoma with a prominent blastemic component revealing a positive reaction for neuron-specific enolase in blastemic and tubular components. Neuron-specific, enolase-positive, small malignant cells are not diagnostic of neuroblastoma when coming from a tumor of the retroperitoneum even if rosettes are present. PMID- 3032540 TI - Control of glycogen synthesis in health and disease. AB - Investigations in our laboratory have shown that the activity of glycogen synthase phosphatase in the liver is shared by at least two functionally distinct proteins: a G-component, which is tightly associated with glycogen particles, and a soluble S-component. Most preparations of glycogen synthase-b that are isolated from the liver of fed glucagon-treated animals require the presence of both components in order to be converted to synthase-a. The G-component is subject to control mechanisms that do not affect the S-component. Its activity is strongly inhibited by phosphorylase-a. This feature explains why glycogen synthesis and glycogenolysis do not normally occur simultaneously, except in the glycogen depleted liver, where a futile cycle may occur. Experiments in vitro have shown that a minimal glycogen concentration is required to ensure the interaction between the G-component and phosphorylase-a. The G-component is also selectively inhibited by Ca2+, and the magnitude of this inhibition depends markedly on the glycogen concentration. The latter inhibition is probably one of the mechanisms by which cyclic adenosine monophosphate (cAMP)-independent glycogenolytic agents achieve the inactivation of glycogen synthase in the liver. Glucocorticoid hormones and insulin are required for the induction and/or maintenance of the G component in the liver. During the development of the fetal rat, glucocorticoids induce the G-component in the liver. This is an essential event in the glucocorticoid-triggered deposition of glycogen in the fetal liver. A functional adrenal cortex is also required in the adult animal to prevent a loss of the capacity for hepatic glycogen storage during starvation. The latter capacity depends on the concentration of functional G-component in the liver. Chronic diabetes causes a similar functional loss. However, the effect of glucocorticoids is not mediated by a putative secretion of insulin. PMID- 3032541 TI - Mechanisms of hormonal regulation of hepatic glucose metabolism. AB - Acute hormonal regulation of liver carbohydrate metabolism mainly involves changes in the cytosolic levels of cAMP and Ca2+. Epinephrine, acting through beta 2-adrenergic receptors, and glucagon activate adenylate cyclase in the liver plasma membrane through a mechanism involving a guanine nucleotide-binding protein that is stimulatory to the enzyme. The resulting accumulation of cAMP leads to activation of cAMP-dependent protein kinase, which, in turn, phosphorylates many intracellular enzymes involved in the regulation of glycogen metabolism, gluconeogenesis, and glycolysis. These are (1) phosphorylase b kinase, which is activated and, in turn, phosphorylates and activates phosphorylase, the rate-limiting enzyme for glycogen breakdown; (2) glycogen synthase, which is inactivated and is rate-controlling for glycogen synthesis; (3) pyruvate kinase, which is inactivated and is an important regulatory enzyme for glycolysis; and (4) the 6-phosphofructo-2-kinase/fructose 2,6-bisphosphatase bifunctional enzyme, phosphorylation of which leads to decreased formation of fructose 2,6-P2, which is an activator of 6-phosphofructo-1-kinase and an inhibitor of fructose 1,6-bisphosphatase, both of which are important regulatory enzymes for glycolysis and gluconeogenesis. In addition to rapid effects of glucagon and beta-adrenergic agonists to increase hepatic glucose output by stimulating glycogenolysis and gluconeogenesis and inhibiting glycogen synthesis and glycolysis, these agents produce longer-term stimulatory effects on gluconeogenesis through altered synthesis of certain enzymes of gluconeogenesis/glycolysis and amino acid metabolism. For example, P-enolpyruvate carboxykinase is induced through an effect at the level of transcription mediated by cAMP-dependent protein kinase. Tyrosine amino-transferase, serine dehydratase, tryptophan oxygenase, and glucokinase are also regulated by cAMP, in part at the level of specific messenger RNA synthesis. The sympathetic nervous system and its neurohumoral agonists epinephrine and norepinephrine also rapidly alter hepatic glycogen metabolism and gluconeogenesis acting through alpha 1-adrenergic receptors. The primary response to these agonists is the phosphodiesterase mediated breakdown of the plasma membrane polyphosphoinositide phosphatidylinositol 4,5-P2 to inositol 1,4,5-P3 and 1,2-diacylglycerol. This involves a guanine nucleotide-binding protein that is different from those involved in the regulation of adenylate cyclase. Inositol 1,4,5-P3 acts as an intracellular messenger for Ca2+ mobilization by releasing Ca2+ from the endoplasmic reticulum.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3032542 TI - Liver glycogen metabolism: an overview. PMID- 3032543 TI - [Interaction of human embryo cells derived from different tissues with lymphoblastoid cell lines]. AB - Cells from skin and muscle of two month human embryo and from nasopharynx, tonsil, lung and kidney of four month human embryo were cocultivated with lymphoblastoid cells--B95-8 and Raji. It is found that all the epithelial cells, such as the focal growth of epithelioid cells from the tonsil and nasopharynx as well as kidney cells from the human embryo can not adhere to any kind of lymphoblastoid cells; but fibroblasts from muscle and skin of two month embryo or from nasopharynx, tonsil and lung of four month embryo have the ability of sticking to lymphoblastoid cells. However, only the fibroblastoid cells from the nasopharynx of human embryo have the ability to stick and to fuse with lymphoblastoid cells--B95-8 in cocultivation and to activate the EBV production. The mechanism of this phenomenon and its biological functions will be studied in detail. PMID- 3032544 TI - [Prostaglandin E (PGE) and gastric carcinoma]. AB - Since 1984, an experimental and clinical study on the relation between PGE and gastric carcinoma has been performed by determining PGE content in the bioptic gastric mucosa and plasma. It is found the PGE content in the gastric mucosa and plasma is increased in all patients with gastric cancer, especially with signet ring cell carcinoma. It is higher in the regional lymph node metastasis than in the early cancer, extensive metastases and normal subjects. The PGE content in the plasma is reduced obviously 7-10 days after operation but is increased markedly in recurrent patients. There is no significant difference in extensive metastases, relapse free and normal subjects. The PGE content in the plasma is significantly higher in gastric carcinoma than in chronic atrophic gastritis, but no difference is present between chronic atrophic gastritis and normal subjects. PMID- 3032545 TI - [Nuclear ultrastructure and histocytologic features of thyroid papillary carcinoma]. AB - Nuclear ultrastructure and histo-cytologic features of 5 cases of thyroid papillary carcinoma were studied. Three main ultrastructural features were: (1) the complex folds and invaginations of the nuclear envelope with formation of nuclear lobules and tiny bridges, (2) the intranuclear pseudoinclusions and (3) the abundant euchromatins. Under the light microscope, as mentioned above in (1), the findings were observed as the indentation of nuclear envelope and deep nuclear groove, (2) was shown as the intranuclear eosinophilic globular body and (3) as the ground-glass appearance of nucleus. The intranuclear eosinophilic globular bodies could be seen on the routine and frozen sections as well as smears. The intranuclear pseudoinclusions and eosinophilic globular bodies, were found in these 5 papillary carcinomas. At the same time, the ultrastructure and histologic features of the follicular cells were observed in 13 cases of colloid adenomas and 9 nodular goiters. None of the follicular cell nuclei showed the three features described above. The intranuclear pseudoinclusion or eosinophilic globular body was not found in any of the ultrathin and histologic sections. These results indicate that in the pathologic diagnosis of thyroid papillary carcinoma, the light microscope has more practical value than the electron microscope. Still does it, especially in the cytologic diagnosis of thyroid puncture before operation. PMID- 3032546 TI - [Double primary cancer in the larynx and lung--report of 12 cases]. AB - 12 patients with double primary cancers in the larynx and lung were treated from 1958 to 1984. The incidence is 1.2% (12/943) of laryngeal carcinomas, 1.3% (12/904) of multiple primary cancers, 1.4% (12/873) of double primary cancers and 39% (12/31) of double primary cancers related to laryngeal cancers. There were 9 male and 3 female. 11 of the first primary cancers occurred in the larynx and only one in the lung. All were proved to be squamous cell carcinoma. In the 11 patients whose second primaries occurred in the lung, 4 were proved to be squamous cell carcinoma, one adenocarcinoma, one oat cell carcinoma, one poorly differentiated carcinoma and one cancer unclassified. Of 10 patients in whom both the first and the second primary cancer were treated, 6 survived for more than 2 years, 4 for 3 years and one for 5 years after the second treatment. It seems that double primary cancers of the larynx and lung could yield favorable results. PMID- 3032547 TI - [Carcinoma of salivary glands--a clinical analysis of 342 cases]. AB - 342 cases with carcinoma of salivary glands were analysed. 152 tumors were located in parotid, 42 in submaxillary, 17 in sublingual and 131 in minor salivary glands. Pathological diagnosis were 106 mucoepidermoid carcinoma, 80 adenoid cystic carcinoma, 54 malignant mixed tumor, 40 adenocarcinoma, 38 papillary cystadenocarcinoma, 17 acinic cell carcinoma, 5 squamous cell carcinoma, and 2 undifferentiated carcinoma. The 3, 5, 10 and 15 year survival rates of these 342 cases were 76.6%, 65.9%, 48% and 29%, respectively. The difference between survival rate and relapse-free survival rate was about 8%. The prognosis of acinic cell carcinoma and mucoepidermoid carcinoma was much better than that of squamous cell carcinoma, adenocarcinoma and undifferentiated carcinoma. The survival rates, according to location, were: minor salivary gland tumors the highest, and those of submaxillary gland tumors the lowest. Postoperative radiotherapy improved the survival rate of adenoid cystic carcinoma. The overall recurrence rate was 37.4%, the neck lymph node metastasis rate 14.3% and the distant metastasis rate 9.1%. PMID- 3032548 TI - Glucose turnover in insulinoma--a case report. AB - To define glucose flux in a state of chronic endogenous insulin excess, a patient with an insulinoma was studied. Plasma glucose, insulin (IRI), glucagon (IRG) and glucose turnover ([3-3H]glucose infusion) were measured before and after insulinoma resection in the postabsorptive state (PA), during a glucose infusion adjusted to attain euglycemia (before insulinoma resection only) and following an intravenous glucagon bolus (1 mg). Before insulinoma resection, plasma glucose was 55 mg/dl, glucose production (Ra) and disappearance (Rd) were equal (1.6 mg/kg/min) and glucose clearance was elevated (2.8 ml/kg/min) in PA. When glycemia was raised with a glucose infusion to 77 mg/dl, Rd did not change; in contrast Ra dropped to zero. Plasma IRI and IRG concentrations were 0.7 ng/ml and 110 pg/ml respectively before glucose infusion and remained constant throughout. After resection of the insulinoma, glycemia in PA was 103 mg/dl, Ra and Rd were increased slightly to 1.9 mg/kg/min while the metabolic clearance of glucose was decreased by 25% (2.1 ml/kg/min). Glucagon stimulation pre- and postinsulinoma resection resulted in significant increases in glycemia and IRI. We conclude that hypoglycemia with insulinoma is a consequence of decreased glucose production and increased glucose clearance. Hepatic sensitivity to small increments in glycemia is markedly enhanced so as to fully suppress endogenous glucose production at euglycemic levels in the absence of any change in IRI and IRG. The mechanisms controlling hepatic Ra in insulinoma appear different from normal. PMID- 3032549 TI - Reversal of diabetes by syngeneic transplantation of a radiation-induced rat insulinoma. AB - The growth and metabolic effects of a radiation-induced rat insulinoma were examined after subcutaneous subscapular transplantation into normal and streptozotocin diabetic NEDH rats. Streptozotocin diabetic rats exhibited hyperglycaemia, hypoinsulinaemia, impaired glucose tolerance without an insulin response, polyuria, polydipsia, hyperphagia and weight loss. Transplantation of tumour fragments gradually improved the physical and metabolic state over the following 3 weeks. Coincident with a progressive rise in plasma insulin between 10 and 17 days, the diabetic rats gained weight and reduced their food intake. The rats remained hyperglycaemic during this time, but developed hypoglycaemia with marked hyperinsulinaemia by 24 days. Furthermore, plasma glucose and insulin concentrations were not increased by an intraperitoneal glucose challenge, indicating greatly accelerated glucose clearance. Both the streptozotocin-treated and normal insulinoma-bearing rats incurred a fatal hypoglycaemic coma by 28-33 days after transplantation. Final body weights, tumour weights and concentrations of glucose and insulin were similar in the two groups. This study demonstrates reversal of streptozotocin diabetes by insulinoma transplantation. The hyperglycaemia and the accompanying diabetic environment did not modify tumour growth and development. PMID- 3032550 TI - [Accuracy of insertion of the Drosophila melanogaster mobile element MDG1 into the cytologically detected "hot spot" of its localization]. PMID- 3032551 TI - [Ultrastructural analysis of the frog sensorimotor synapse physiologically identified and labeled with horseradish peroxidase]. PMID- 3032552 TI - Synergy of sulbactam and ampicillin against methicillin-resistant staphylococci. AB - Methicillin resistance in staphylococci is an increasing problem both for Staphylococcus aureus (MRSA) and Staphylococcus epidermidis (MRSE), which cause infections of the heart and after central nervous system surgery. Resistance seems to be due primarily to production of altered penicillin-binding proteins. The present study determined whether a combination of beta-lactamase inhibitor sulbactam and ampicillin or sulbactam and cefazolin would inhibit MRSA and MRSE. Sulbactam, ampicillin and cefazolin at 32 micrograms/ml did not inhibit MRSA or MRSE. At 8 micrograms/ml of each agent all isolates were inhibited. Synergy of sulbactam and ampicillin could be demonstrated against MRSA by the agar fixed ratio method, checkerboard dilution and by killing curves. This suggests that in certain situations MRSA and MRSE may be effectively eliminated by this method. PMID- 3032554 TI - [Myocardial scintigraphy for the early evaluation of the success of thrombolysis therapy in acute myocardial infarct]. PMID- 3032555 TI - [Review: Acquired immunodeficiency syndrome of man (AIDS) and of primates (SAIDS)]. PMID- 3032553 TI - Membrane phospholipids alter nutrient transport and drug toxicity in tumorigenic fibroblasts. AB - The phospholipid polar head group composition of LM cell plasma membranes was nutritionally altered by choline analogue supplementation. Phosphatidylcholine (PC), normally 60% of total phospholipid, was depleted by 60 to 90% in membranes from cells cultured with N, N'-dimethylethanolamine (DME), N monomethylethanolamine (ME), and ethanolamine (E). Enrichment of LM cell membranes with choline analogues, such as DME-, ME-, and E-containing phospholipids, decreased the transport of [3H]thymidine, [3H]2-deoxy-D-glucose, and [14C]3-O-methylglucose. Conversely, no change in the transport of [3H] uridine or [14C]aminoisobutyric acid was observed. The toxicity of antineoplastic drugs such as 5-fluorouracil, but not daunorubicin, doxorubicin, or methotrexate, was enhanced threefold in cells enriched with phospholipid containing choline or DME as compared to ME or E. Arrhenius plots of Na+-K+-ATPase activity demonstrated a characteristic temperature at 29 degrees C in plasma membranes from choline-fed cells, while those from analogue-fed cells showed an additional break at 20 degrees C and had higher energies of activation below this temperature. In addition, choline analogue supplementation altered the protein composition of the plasma membrane. The results reported herein demonstrate that nutritional alteration of LM fibroblast plasma membrane phospholipid polar head group composition affects several transport processes and toxicity of some anticancer drugs. PMID- 3032556 TI - [Infectious course of equine herpesvirus 1 infection in a riding stable]. PMID- 3032557 TI - [Influenza epidemic in horses in West Berlin 1983-1985. 1. Clinical and hematological findings]. PMID- 3032558 TI - [Influenza epidemic in horses in West Berlin 1983-1985. 2. Virological and serological findings]. PMID- 3032559 TI - [Bovine viral diarrhea (BVD) as the cause of calf mortality among cattle breeding stock in the Wesermarsch district]. PMID- 3032560 TI - Symposium on angiotensin-converting enzyme inhibition in cardiac failure. Oslo, 23 October 1985. Proceedings. PMID- 3032561 TI - Clinical pharmacology of the ACE inhibitors. AB - The central role of the renin-angiotensin-aldosterone system in the regulation of fluid balance and haemodynamics was not fully appreciated until the discovery and clinical application of inhibitors of the angiotensin-converting enzyme (ACE). Captopril, the first orally active compound has proved to be highly effective in mild, intermediate and severe hypertension and in congestive heart failure. This is also true for enalapril, the first of the second generation (non-sulfhydryl) of ACE inhibitors. Both compounds combine a high degree of clinical efficacy with a low rate of side effects, and both are eliminated via renal excretion. Thus, reduced doses are required in renal failure. Captopril has a shorter half-life and requires more frequent dosing than enalapril. Whether long-acting ACE inhibitors carry distinct advantages or disadvantages compared with short-acting ones is not clear, but both possibilities must be considered. Among many new ACE inhibitors being developed, compounds eliminated via hepatic routes, such as fosfenopril and zofenopril, may prove advantageous in renal failure. Very long acting ones, such as lisinopril, a potent ACE inhibitor already shown to be clinically effective, may add value to this group of therapeutic agents. Drug compliance may be easily tested by measuring ACE activity in serum from patients treated with stable ACE inhibitors, such as enalapril and lisinopril, which is an obvious advantage compared with other antihypertensive compounds. The presence of ACE in sites not associated with regulation of fluid balance or blood pressure, such as macrophages and reproductive organs, and the possibility that new functions of ACE may be discovered, call for vigilance regarding possible long term side effects of ACE inhibitors. PMID- 3032563 TI - Effects of enalapril on different aspects of the clinical state in congestive heart failure. AB - In patients with moderate to severe CHF, enalapril improves symptoms and is well tolerated. It increases functional capacity in the form of exercise time and reduces the left ventricular dimensions. Enalapril also has favourable effects on electrolytes and, although it increases renal blood flow, it reduces the glomerular filtration rate. PMID- 3032562 TI - Pharmacokinetics of enalapril in congestive heart failure. AB - The pharmacokinetics of the converting enzyme inhibitor enalapril were studied in an open, randomised, balanced crossover design in 12 hospitalised patients with stable, chronic congestive heart failure (CHF). Enalapril maleate is a prodrug requiring in vivo hepatic esterolysis to yield the active diacid inhibitor enalaprilat. CHF results in changes in regional blood flow that may affect the gastrointestinal absorption, hepatic hydrolysis and renal excretion of enalapril and enalaprilat. In order to evaluate the pharmacokinetics of enalapril in CHF, the following treatments were given: enalapril 10 mg orally, enalapril 5 mg intravenously and enalaprilat 5 mg intravenously. Each dose was followed by a 72 hour period with frequent blood sampling and fractionated urine collection for the radioimmunoassay of both enalapril and enalaprilat. Mean absorption for the oral dose was 69%, hydrolysis 55%, bioavailability 38%, urinary recovery 77% and estimated first-pass effect 10%. The results were compared with available data in normal subjects. After oral administration of 10 mg enalapril, the extent of absorption, the degree of hydrolysis and the bioavailability in CHF patients appear to be similar to those in normal subjects, with differences less than 10%. The rates of absorption and hydrolysis appear to be slightly slower in CHF. The serum concentrations of enalaprilat were consistently greater in CHF, and maximal concentrations were reached at 6 hours in CHF compared with 4 hours in normal subjects. The maximal hypotensive responses were similar for all three treatments, although the onset of action was rapid following intravenous enalaprilat. It is concluded that the presence of CHF does not appreciably alter the pharmacokinetic behaviour of enalapril.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3032564 TI - When to use ACE inhibitors in heart failure. PMID- 3032565 TI - [The defect patient in the post-hospitalization stage]. PMID- 3032566 TI - [Surgical treatment possibilities for periodontally and apically diseased teeth]. PMID- 3032567 TI - Role of postnatal gonadal function in the determination of thyrotropin (TSH) releasing hormone-induced TSH response in adult male and female rats. AB - Permanent effects of postnatal gonadal function on the hypothalamo-pituitary thyroid axis were examined in male and female rats of the Wistar-Imamichi strain. Animals were used at the age of about 10 weeks. Neonatally castrated (NC) males showed a significantly higher plasma TSH response to TRH (10 micrograms/kg BW, ip) than males castrated at 1, 2, 3, 4, or 6 weeks of age. The TSH response in intact males was similar to that seen in NC males. Neonatal androgenization of NC males with 100 micrograms/rat of testosterone propionate decreased the TSH response in the group. The anterior pituitary (AP) TSH content was similar among all male groups. The number of AP TRH receptors was significantly higher in both NC males and intact males than in both NC + neonatal androgenization males and 6 week-castrated males. In female rats, TRH-induced TSH response, AP TSH content, and AP TRH receptor number were all unaffected by the age of castration, whereas intact females significantly exceeded 6-week-castrated females in these three variables. The circulating levels of T3 and T4 and hypothalamic TRH content were similar in all groups in each sex. These results indicate biphasic effects of postnatal testosterone on the TSH response to TRH, that is, a permanent inhibition in early postnatal days and a transient stimulation in adult age. The effects occurred not via changes in AP TSH content but via changes in AP TRH receptor number. In females, however, postnatal gonads did not exert any permanent effects on the TRH-TSH system. PMID- 3032569 TI - Presence of gonadotropin-releasing hormone-like proteins in bovine and ovine ovaries. AB - Recently, the rat ovary was shown to contain significant levels of a GnRH-like protein, but no detectable GnRH. In the present studies, extracts of bovine ovaries and ovine corpora lutea were examined for their content of both GnRH-like protein and GnRH. The GnRH-like proteins were detected with a rat ovarian membrane radioreceptor assay, and GnRH was detected with a specific GnRH RIA. The biological activity of the GnRH-like protein was evaluated with a rat luteal cell assay. GnRH-like activity, but not GnRH, was clearly present in extracts of the entire bovine ovary, bovine corpus luteum, bovine granulosa cells, and ovine corpus luteum. The highest levels of GnRH-like activity were present in granulosa cells. Neither GnRH-like activity nor GnRH was detected in extracts of bovine follicular fluid or bovine jugular plasma. Fractionation of the bovine and ovine GnRH-like proteins by reverse phase HPLC resulted in retention times similar to that of the rat ovarian GnRH-like protein, but distinctly different from that of authentic GnRH. The bovine and ovine GnRH-like fractions, like those in the rat, were sensitive to protease and heat. The bovine GnRH-like protein, obtained by preparative reverse phase HPLC, evoked a dose-dependent inhibition of LH stimulated cAMP accumulation in rat luteal cells similar to that caused by GnRH. Based on these results, we suggest that the bovine ovary, granulosa cells, and corpus luteum and the ovine corpus luteum contain an antigonadotropic GnRH-like protein similar to the GnRH-like protein of the rat ovary. The presence of a similar GnRH-like protein (but the absence of GnRH) in ovaries of domestic species and the rat raises the possibility that this substance may play a paracrine antigonadotropic role in the ovary of diverse species. PMID- 3032568 TI - Growth hormone-releasing factor-44 specificity for components of somatotroph and lactotroph immediate release pool substructures. AB - Rat somatotroph and lactotroph hormone storage is divisible into at least two functional compartments: an immediate release pool (IRP) and a pool that responds to prolonged stimulation. An IRP substructure has been defined by release in response to potassium ion (K+), prostaglandin E1 (PGE1), and Bu2cAMP. The somatotroph IRP is expandable; the lactotroph IRP is fixed in size. The present experiments examined which IRP components contribute to the rapid release of stored GH in response to GH-releasing factor-44 (GRF). Release of stored PRL was monitored for comparison. In vitro prelabeling defined stored rat (r) GH and rPRL. Release in response to 21 mM K+, 3 microM PGE1, 1 mM Bu2cAMP, and/or 3 nM GRF was monitored with a perifusion-immunoprecipitation system. After 120 min of basal perifusion, tissue was exposed to one of the four secretagogues for 90 min. During a second 90-min period a second secretagogue was added while exposure to the first secretagogue continued. We demonstrated that 21 mM K+ reduces peak rGH release in response to 3 nM GRF by 52%, whereas GRF does not reduce rGH release in response to K+; 3 microM PGE1 reduces rGH release in response to GRF by only 19% although GRF reduces rGH release in response to PGE1 by 88%; 1 mM Bu2cAMP reduces rGH release in response to GRF by 87%, and GRF eliminates rGH release in response to Bu2cAMP (1.2% of control value); combined K+ plus Bu2cAMP reduce rGH release in response to GRF to 2.5% of the control value, whereas after GRF pretreatment rGH release in response to combined K+ plus Bu2cAMP is 93% of the control value; and combined PGE1 and Bu2cAMP reduce the response to GRF to 17% of the control value. Effects on rPRL release are qualitatively similar. We conclude that immediate GRF-stimulated release of stored rGH originates in the somatotroph IRP components defined by responses to PGE1 and Bu2cAMP; it derives only slightly, if at all, from the IRP component defined by the response to K+. The smaller GRF-stimulated release of IRP rPRL is similarly derived. PMID- 3032570 TI - Chronic morphine treatment enhances the negative and positive feedback effects of estradiol on gonadotropin secretion in ovariectomized rats. AB - Studies were undertaken to evaluate the effects of chronic morphine exposure on the negative and positive feedback effects of estradiol (E2) on LH and FSH secretion in ovariectomized rats. Two days of E2 exposure reduced at 1000 h and stimulated at 1600 h the serum LH concentration at each dose tested. Chronic exposure (4 days) to morphine enhanced both the inhibitory effects of E2 on LH secretion at 1000 h and E2 induction of the midafternoon surge in LH. A detailed time course of the afternoon LH response to E2 revealed that chronic morphine exposure advanced its time of onset and increased its magnitude. The response of FSH to E2 and the interaction between E2 and morphine on FSH secretion were similar to those observed for LH, albeit lower in magnitude. With extension of the E2 exposure period from 2 to 4 days, the synergistic interaction between E2 and morphine persisted for the negative, but not the positive, feedback action on LH. Chronic morphine exposure in ovariectomized rats, without E2 replacement, was ineffective in altering the LH levels at 1000 h, but had a slight stimulatory effect on LH at 1600 h, indicating an E2 requirement for the observed suppression and stimulation of LH in the morning and afternoon, respectively. The response of the anterior pituitary to exogenously administered LHRH in the morning (1000 h) and during the midafternoon hypersecretion of LH (1800 h) was markedly enhanced in animals exposed chronically to morphine. Thus, the chronic stimulation of opiate receptors with morphine enhances the inhibition and subsequent hypersecretion of LH in E2-exposed rats, advances the time of onset and enhances the magnitude of the E2-induced LH surge, and augments the response to LHRH when LH levels are low in the morning and during the E2-induced hypersecretion of LH. These data support an important role for opioid neuronal systems in the mechanism that switches the E2 signal for gonadotropin secretion from inhibitory to stimulatory and, hence, in the series of neuronal events that leads to phasic secretion of gonadotropins. PMID- 3032571 TI - Control of cyclic adenosine 3',5'-monophosphate production in osteoclasts: calcitonin-induced persistent activation and homologous desensitization of adenylate cyclase. AB - Hormonal control of cAMP production in the osteoclast has not been investigated in detail because this bone-resorbing cell has been difficult to isolate. We have used osteoclasts freshly isolated by disaggregation from neonatal rat long bones and settled onto coverslips (100-150 cells per coverslip) to examine the effects of calcitonin and prostaglandin E2 on osteoclast cAMP levels and cytoplasmic spreading. Salmon, eel, and human calcitonin (CT), and various analogs, stimulated cAMP production in a dose-dependent manner with relative potencies as seen in other response systems. Forskolin (10(-7) M) increased the sensitivity and amplitude of the response. Pretreatment with pertussis toxin (200 ng/ml for 3 h) had no effect suggesting that CT does not act through Ni, the inhibitory guanine nucleotide regulatory unit of adenylate cyclase. CT treatment was associated with rapid and dose-dependent induction of a persistent activated state of adenylate cyclase and homologous desensitization, the former being a particular feature of CT action previously observed in nonosteoclastic cells. Quantitative histomorphometry demonstrated a sensitive, dose dependent, and prolonged (greater than 2 h) reduction in osteoclast plan area after exposure to salmon CT. Although prostaglandin E2 also stimulated cAMP production and resulted in cell contraction in osteoclasts this was not associated with persistent activation of adenylate cyclase nor with prolonged contraction. Persistent activation of adenylate cyclase may be an important mechanism in CT inhibition of osteoclast function. PMID- 3032572 TI - Ultrastructural localization of cyclic adenosine 3',5'-monophosphate-dependent protein kinase after adrenocorticotropin stimulation in adrenal cortical tumor cells. AB - To increase our knowledge of the molecular details of peptide hormone action, a specific immunogold staining procedure for the ultrastructural localization of cAMP-dependent protein kinase regulatory (RI) and catalytic (C) subunits was used in Y-1 adrenal cortical tumor cells. The Y-1 adrenal cell responds to ACTH (40 mU/ml) with a decrease in cell division and an increase in steroid production. Corresponding to the decrease in rate of cell division and the increase in steroid production, there was a 2-fold increase in both nuclear and cytoplasmic localization of the C subunit after 60-min ACTH (40 mU/ml) treatment of the culture. The amount of immunogold staining in the adrenal tumor cells after localization with antiserum of the RI subunit decreased after 60-min ACTH (40 mU/ml) stimulation. A 2-fold decrease in labeling in the cytoplasm and nucleus was observed for the RI subunit. The loss of RI subunit from the soluble fraction of the cytoplasm of the cell or an alteration of this RI subunit is suggested by this investigation. Since there was no increase in the RI subunit in the nuclear compartment, a loss of RI subunit from the cytoplasm into the nucleus seems unlikely. The observed immunogold changes in the C subunit after ACTH treatment correspond to the reported changes observed with light microscopic techniques with a fluorescein-coupled inhibitor as a probe for the localization of free C. The immunogold technique allows for the ultra-structural identification and quantification of nuclear as well as the cytoplasmic sites of cAMP-dependent protein kinase after hormonal stimulation. These results support the proposed role of protein kinase as a mediator of the ACTH response. PMID- 3032573 TI - Tissue distribution and hormonal regulation of messenger ribonucleic acid for regulatory and catalytic subunits of adenosine 3',5'-monophosphate-dependent protein kinases during ovarian follicular development and luteinization in the rat. AB - cDNA probes specific for the regulatory (R) subunits [RII51; (mol wt, 51,000) and RI (mol wt, 49,000)] and a catalytic (C alpha) subunit of cAMP-dependent protein kinases were used to analyze the hormonal regulation, tissue distribution, and content of mRNAs for these kinase subunits in the rat ovary. Filter hybridization assays demonstrated that mRNA specific for RII51 increased in both thecal and granulosa cells of preovulatory (PO) follicles, then declined precipitously (less than 10-fold) in both cell types within 7 h after an ovulatory (10-IU) dose of hCG and remained low in corpora lutea. Dose-response studies showed that doses of FSH greater than 2 micrograms were required to increase RII51 mRNA in granulosa cells of hypophysectomized (H) rats, whereas doses of 0.5-1.0 micrograms were effective in granulosa cells of H estradiol-treated (HE) rats. At all doses (0.5 50 micrograms) of FSH administered, RII51 mRNA was 4 times higher in granulosa cells of HE rats than in H rats. Solution hybridization assays demonstrated that the concentration of RII51 mRNA increased 6-fold from 20 molecules/granulosa cell in H rats to 120 molecules/cell in H rats treated with estradiol and FSH (HEF). Transcription assays using nuclei of granulosa cells demonstrated further that the 5- to 10-fold increases in RII51 mRNA content in estradiol-/FSH-induced granulosa cells was associated with increased transcription of the RII51 gene. In thecal cells of small antral (SA), PO, and luteinizing follicles, changes in the content of mRNA for RI and C alpha kinase subunits showed a pattern similar to that for RII51. However, in granulosa cells, mRNA specific for RI and C alpha was highest in SA follicles, declined in PO follicles, and remained unchanged during luteinization. The content of mRNA for RI (45-70 molecules/cell) and C alpha (10 molecules/cell) also changed less than 2-fold in granulosa cells of H, HE, and HEF rats. In summary, these results indicate that the content of RII51 mRNA is hormonally regulated in both thecal cells and granulosa cells during follicular development and luteinization. Low concentrations of gonadotropins increase RII51 mRNA, whereas an ovulatory dose of hCG causes mRNA for RII51 to decrease rapidly. In granulosa cells, induction of mRNA for RII51, but not that for RI and C alpha is induced by the actions of estradiol and FSH, and involves increased transcription of the RII51 gene. PMID- 3032574 TI - Hormonal regulation of the production of collagenase and a collagenase inhibitor activity by rat osteogenic sarcoma cells. AB - Collagenases that specifically cleave native collagen at neutral pH have been implicated in the maintenance and turnover of connective tissue. In bone, the origin of neutral collagenase has remained equivocal, although recent studies have indicated that it is synthesized by the osteoblast. In the present work, regulation of secretion of neutral collagenase and a collagenase inhibitory activity was investigated using the osteoblastic tumor cell line UMR 106-01 and a variety of bone-resorbing agents. Under basal conditions, UMR 106-01 cells produced very low levels of collagenase but substantial amounts of the inhibitory activity. Exposure to PTH and, to a lesser extent, 1,25-dihydroxyvitamin D3, prostaglandin E2, retinoic acid, and epidermal growth factor stimulated the release of collagenase, an effect not seen with interleukin-1 or heparin. The stimulation of collagenase by PTH was dose dependent, with a half-maximal response occurring at 10(-8) M. Inclusion of isobutylmethylxanthine decreased the concentration of PTH required to produce half-maximal stimulation to 2 X 10(-10) M, indicating action via cAMP. With respect to the inhibitory activity, PTH and epidermal growth factor were the only agents, among those tested, able to enhance its production. Both hormones caused a 50-100% increase over control levels 72 h after hormone administration. There were notable differences in the time courses of production of collagenase and the inhibitor. After treatment with PTH, the enzyme reached maximal concentrations between 12-48 h, but declined to undetectable levels by 96 h. In contrast, the inhibitory activity was secreted in a linear fashion, with the highest concentrations achieved around 72-96 h. These results suggest a complex pattern of regulation of collagenase and inhibitor secretion by the osteoblastic cell, with the steady accumulation of inhibitor perhaps being responsible for the ultimate curtailment of enzyme activity. PMID- 3032575 TI - Age-related change in ketone body metabolism: diminished glucagon effect on ketogenesis in adult rats. AB - The age-related changes in plasma ketone body levels and related substances such as carnitine and FFA of normal and streptozotocin diabetic rats, as well as its effects on glucagon-induced ketogenesis in isolated perfused rat livers, were examined in this study. The degree of increase of acetoacetate (AcAc) in adult rats (50-week-old) after a 36-h fasting period in both normal and diabetic rats was significantly smaller than that of young rats (8-week-old). Plasma total carnitine tended to decrease with aging. On the other hand, the plasma levels of FFA and glucagon in fasted adult rats were significantly higher than those in young rats. In parallel with the in vivo observation, the basal output of AcAc, but not 3-hydroxybutyrate, from adult rat livers was significantly smaller than that of the young normal and diabetic rats. The level of glucagon-stimulated AcAc output from the young rat liver was significantly higher than that from the adult rat liver in both the normal and diabetic rats. This study demonstrates that the hepatic unresponsiveness to glucagon in terms of its ketone body production by aging may be one of the major causes of hyperosmolar nonketotic coma in elderly people. PMID- 3032576 TI - Regression of renal hypertrophy and elevated renal Na+,K+-ATPase activity after insulin treatment in streptozotocin-diabetic rats. AB - The effect of insulin treatment on the renal hypertrophy and elevated renal Na+,K+-ATPase activity in rats with streptozotocin (STZ)-induced diabetes was examined. Rats with STZ-diabetes of 6- to 8-week duration had significantly lower body weights, higher plasma and urinary glucose concentrations, greater urinary volumes, increased kidney weights, and increased kidney/body weights and protein/kidney weight ratios compared to those in saline-citrate-injected controls. Specific Na+,K+-ATPase activity per mg protein in both cortical and outer medullary kidney homogenates was significantly elevated in diabetic vs. control animals, as was total renal Na+,K+-ATPase activity. One week of insulin treatment returned elevated plasma glucose, urinary volume, the protein/kidney weight ratio, and cortical and outer medullary Na+,K+-ATPase activity per mg protein to control values. Kidney weights and kidney/body weight ratios of diabetic animals remained elevated, as did absolute total renal Na+,K+-ATPase activity. After 3 weeks of insulin treatment, kidney weight and total renal Na+,K+-ATPase activity in diabetic animals returned to control values, but body weights remained lower than those in the controls, resulting in continued elevation of kidney/body weight ratios in the diabetic animals. The concurrent regression of both renal hypertrophy and elevated Na+,K+-ATPase activity to normal levels after insulin treatment of STZ-diabetic animals implicates renal growth rather than a direct effect of insulin as the primary factor controlling elevation and regression of Na+,K+-ATPase activity in the diabetic kidney. This finding demonstrates that the effect of renal hypertrophy can outweigh the intrinsic effects of insulin on an important renal transport system and that this effect may be as important as lack of hormone in determining the renal physiological responses in the disease. It is suggested that the increased renal tubular Na+,K+-ATPase activity is a key component of the renal hypertrophy and hyperfunction seen in diabetes. PMID- 3032577 TI - Hormonal and immunological characterization of the cell-associated plasminogen activators produced by cultured rat granulosa cells. AB - The hormonal induction and immunological reactivities of the cell-associated plasminogen activators (PAs) produced by granulosa cells obtained from the ovaries of diethylstilbestrol-implanted immature rats were studied. FSH and other cAMP-inducing ligands, including cholera toxin, forskolin, and 8-bromo-cAMP, elevated the PA activity of granulosa cells in a concentration-dependent manner during a 4-h culture, with an approximate 10-fold maximal increase in PA activity compared to control cells. Negligible levels of PA activity were observed in the extracellular medium in the absence or presence of hormones. The PA induced by FSH or cAMP in intact cells was progressively neutralized during the 4-h culture by increasing amounts of antibodies to the tissue-type PA (tPA), but not by an IgG fraction against the urokinase-type PA (UK-PA). However, solubilization of granulosa cells with Triton X-100 revealed the presence of intracellular UK-PA activity in both FSH-treated and control cells that consisted of about 20% of the total cellular PA activity. Electrophoretic analysis of extracts from solubilized granulosa cells indicated the presence of three peaks of PA activity. A PA with a Mr of 70,000 was induced by FSH, was completely inactivated by tPA antibodies, and required fibrin for full activity. A 40,000 Mr fibrin-independent PA was also stimulated by FSH and was partially inhibited by UK-PA antibodies, but not by anti-tPA immunoglobulin G. A third peak of PA activity comigrated with a human UK PA standard at a Mr of 33,000, was fully neutralized by UK-PA antibodies, and was largely present only in control cells. These results suggest that during the first hours of granulosa cell development, FSH via cAMP induces the production of a cell-surface tPA, while both FSH-treated and control cells synthesize intracellular UK-PAs. Hormonal regulation of the production and activities of these cellular enzymes may allow the expression of specific differentiated functions of developing granulosa cells. PMID- 3032578 TI - Effect of relaxin on aromatase activity in human endometrial stromal cells. AB - Previous studies have shown that the aromatase activity in human endometrial stromal cells was stimulated by progestin and enhanced by estrogen and forskolin (Fk), an agent that stimulates the accumulation of intracellular cAMP. Present study was undertaken to investigate whether any peptide hormone would affect endometrial aromatase activity. Stromal cells were isolated from normal proliferative and secretory endometria and cultured in nutrient medium. Porcine relaxin (RLX) was added to culture medium individually or in combination with medroxyprogesterone acetate (MPA) and estradiol (E2). Cells treated with RLX alone did not affect the aromatase activity. RLX, however, exerted a synergistic effect on aromatase activity in the presence of MPA or MPA plus E2. On the other hand, human CG, epidermal growth factor, human PRL, and insulin did not increase the aromatase activity in the presence or absence of MPA and E2 studied in a limited number of specimens. The progestin-dependent effect of RLX on aromatase activity was dose dependent indicating that the biological effect of RLX is mediated through a saturable mechanism. When RLX was added to MPA-pretreated cells, additional increase of aromatase activity was seen after 24 h incubation indicating that the action of RLX on stromal cells is not an acute effect. Antiprogestin, RU486, inhibited the stimulation of aromatase activity in both MPA and MPA plus RLX treated cells. RLX has either no effect or a moderate increase (up to 2-fold over the control) on intracellular cAMP content. On the other hand, Fk increased the intracellular cAMP level and enhanced the aromatase activity in the presence of progestin. Also RLX did not replace the effect of Fk since additional increase of aromatase activity was noted when stromal cells were incubated with MPA plus RLX plus Fk in comparison to MPA plus RLX or MPA plus Fk. These results suggest that the action of RLX on stromal cells may be mediated through an intracellular messenger independent of cAMP. Present studies provide evidence that RLX exerts a synergistic effect on aromatase activity in the presence of progestin in human endometrial stromal cells. It is evident that human endometrium is a target organ of RLX. PMID- 3032579 TI - Alpha 1-adrenergic receptors in the neural lobe of the rat pituitary: autoradiographic identification and localization. AB - alpha 1-Adrenergic receptors were identified, characterized, and localized in rat pituitary gland by quantitative light microscopic autoradiography. Autoradiographic studies were carried out in slide-mounted rat pituitary sections using both [125I]2-[beta-(4-hydroxy-3-iodophenyl)ethyl-aminomethyl]tetralone ([125I]HEAT) and [3H]prazosin to localize alpha 1-adrenergic receptors. Data analysis by densitometry showed that [125I]HEAT binding in the rat neural lobe was saturable and of high affinity, with an apparent dissociation constant (Kd) of about 5 pM. Data from competition studies using a variety of compounds demonstrated an alpha 1-adrenergic receptor profile for [125I]HEAT-binding sites in the rat pituitary. A high density of alpha 1-adrenergic receptors (1 microM prazosin-displaceable [125I]HEAT binding or 10 microM phentolamine-displaceable [3H]prazosin binding) was found present only in the neural lobe, with negligible concentrations in the anterior and intermediate lobes. The regulation of [125I]HEAT-binding sites in the neural lobe was examined in pituitary stalk transected and superior cervical ganglionectomized rats. Significant increases in [125I]HEAT-binding sites were observed after superior cervical ganglionectomy, but no changes in [125I]HEAT binding were found in pituitary stalk-transected rats compared to that in sham-operated controls. These data provide the first identification of alpha 1-adrenergic receptors in the neural lobe of the rat pituitary and suggest that these receptors may be localized primarily in blood vessels. In addition, a primary role for the peripheral sympathetic nervous system in regulating the neurohypophyseal vasculature is suggested. The precise function of alpha 1-adrenergic receptors in the neural lobe in regulating posterior lobe hormone secretion remains to be demonstrated. PMID- 3032580 TI - Effects of epinephrine and norepinephrine on lipid mobilization from coho salmon liver incubated in vitro. AB - The direct effects of epinephrine and norepinephrine (NE) on lipid mobilization were studied in coho salmon liver slices incubated in vitro. Fatty acid (FA) release from liver slices was measured continuously by pH-stat titration. The pH stat assay system was validated by simultaneous colorimetric measurement of FA and glycerol release. Liver slices incubated in a glucose-free, low buffering capacity medium and stimulated with NE (10(-6) M) exhibited a one proton (titrimetric) to one FA (colorimetric) release profile. Under similar conditions, NE administration also stimulated glycerol release, in the expected three FA (colorimetric) to one glycerol (colorimetric) ratio. NE stimulated FA release from liver slices in a dose-dependent manner; epinephrine did not have a lipolytic effect. The beta-agonist isoproterenol stimulated FA release, whereas alpha-agonists had no effect. Furthermore, the beta-antagonist propranolol inhibited both NE- and beta-agonist-stimulated FA release. Liver triacylglycerol lipase activity was also stimulated by NE. Propranolol inhibited NE-stimulated lipase activity. These results establish the presence of hormone-sensitive lipase in salmon liver and suggest that NE-stimulated lipid mobilization in salmon liver is mediated through beta-adrenergic pathways. PMID- 3032581 TI - Compensatory response of the luteinizing-hormone (LH)-releasing hormone (LHRH)/LH pulse generator after administration of a potent LHRH antagonist in the ram. AB - It is established that the blockade of the pituitary LHRH receptor by an LHRH antagonist will suppress pituitary LH secretion and reduce serum concentrations of gonadal steroids. Little is known, however, about the activity of the LHRH/LH pulse generator during this inhibitory period or during the recovery phase. To investigate this, a potent LHRH antagonist [N-Ac-D-pCl-Phe1,D-pCl-Phe2,D-Trp3,D hArg(Et2)6, D-Ala10 LHRH was injected iv into sexually active rams and the changes in the blood plasma concentrations of LH, FSH, testosterone, and PRL were measured in samples collected every 15 min for 24-48 h. The treatment induced an immediate blockade of pulsatile LH secretion and a parallel decline in blood levels of testosterone. Plasma levels of FSH were not suppressed by treatment with the LHRH antagonist and there was no consistent effect on plasma levels of PRL. The duration of the inhibition of LH was dose dependent lasting 4.3 +/- 0.4 h, 18.0 +/- 1.0 h, and 31.8 +/- 1.3 h for the low (6 micrograms/kg), medium (36 micrograms/kg), and high (365 micrograms/kg) doses of LHRH antagonist, respectively. During the recovery period there was an approximate 2-fold increase in the frequency of LH pulses. These results suggest a compensatory response to the decline in the negative feedback effect of testosterone secretion. Even the lowest dose of antagonist elicited a decrease in the level of testosterone and an increase in LH pulse frequency. At this dose, the decline in testosterone was very transitory indicating an acute sensitivity of the hypothalamus to changes in the negative feedback signal. These results suggest that the suppression of LH and testosterone secretion in the ram by LHRH antagonist is associated with a compensatory increase in the activity of the LHRH pulse generator. PMID- 3032582 TI - Interaction of receptors for insulin-like growth factor I, platelet-derived growth factor, and fibroblast growth factor in rat aortic cells. AB - Serum contains various growth factors which regulate the proliferation of cells. We investigated the growth of cultured arterial smooth muscle cells under the influence of insulin-like growth factor I (IGF I), fibroblast growth factor (FGF), and platelet-derived growth factor (PDGF), and examined the effect of these growth factors on the binding of [125I] IGF I and on the binding of [125I]PDGF to these cells. IGF I, FGF, and PDGF stimulated [6-3H]thymidine incorporation into DNA of confluent cultures of cells which were incubated in modified Dulbecco's modified Eagle medium. However, the effect of these growth factors on DNA synthesis was much more potent in Dulbecco's modified Eagle medium with 1% fetal calf serum. FGF and PDGF potentiated the growth-promoting effect of IGF I. The binding of [125I]IGF I to the cells was increased after a preincubation with FGF and PDGF. The binding was potently increased by FGF (100 ng/ml) after a preincubation time of 30 min. There was an increase in binding during the first 3 h of preincubation followed by a decrease after 4-5 h. PDGF (10-1000 ng/ml) stimulated [125I]IGF I binding only after 2 h of preincubation. The stimulation was dose dependent. Maximal stimulation of the binding was observed after 3 h of preincubation followed by a decrease after 4-5 h of preincubation. Specific binding sites for PDGF on smooth muscle cells could be demonstrated too. A preincubation of confluent cells with IGF I caused a dose dependent increase in [125I]PDGF binding. These results support the hypothesis that the regulation of the binding of a specific growth factor by a second growth factor is important for the control of cell growth. PMID- 3032583 TI - Calcium release from porcine thyroid microsomes by phosphatidylinositol 4,5 bisphosphate and inositol 1,4,5-trisphosphate. AB - We have demonstrated Ca2+ mobilization induced by inositol compounds from intracellular stores of isolated porcine thyroid cells that were permeabilized with saponin and from fractionated thyroid microsomes that were preloaded with Ca2+. In the presence of saponin and antimycin A, a mitochondrial inhibitor, the Ca2+ concentration in the cell suspension decreased after the addition of ATP; the addition of inositol 1,4,5-trisphosphate (IP3) elicited transient elevations in Ca2+. The half-effective concentration (EC50) of IP3 was about 1 microM. TSH, acetylcholine, and norepinephrine had no effect on the Ca2+ movement, but phosphatidylinositol 4,5-bisphosphate (PIP2) induced a similar Ca2+ release (EC50 = 3 microM). PIP2-induced release decreased as the Mg2+ concentration was raised. After the release induced by a maximal amount of IP3, PIP2 could induce a further Ca2+ release. We also measured Ca2+ efflux from fractionated microsomes preloaded with 45Ca2+ by ATP-dependent transport. IP3 induced a small efflux of 45Ca2+, but PIP2 was much more effective at low Mg2+ concentrations. More than half of the loaded 45Ca2+ was released by 10 microM PIP2 within 30 sec, and the EC50 was 0.7 microM. These results suggest that Ca2+ mobilization from the microsomes mediated by inositol compounds participates in the regulation of thyroid cell functions. PMID- 3032584 TI - Diminished diurnal secretion of adrenocorticotropin (ACTH), but not corticosterone, in old male rats: possible relation to increased adrenal sensitivity to ACTH in vivo. AB - The diurnal secretion of ACTH and corticosterone was examined in chronically cannulated young (3-4 months old), middle-aged (10-12 months old), and old (22-24 months old) Fischer 344 male rats. Plasma corticosterone in young rats increased from baseline concentrations of 78 +/- 5 to a maximum of 171 +/- 24 ng/ml at 1730 h and declined to basal levels by 1930 h. Middle-aged and old rats demonstrated a similar magnitude and time course of corticosterone release. However, comparison of the relative concentrations of ACTH released during the diurnal surge revealed that old rats secreted 35% less ACTH than young or middle-aged animals (P less than 0.05). Age-related changes in the sensitivity of the adrenal gland to a submaximal dose of ACTH were tested in dexamethasone-pretreated animals at 1100 and 1700 h in a separate experiment. Plasma corticosterone levels were significantly greater after ACTH administration (1 mIU/kg ACTHAR, iv) at 1700 h in both young and old rats compared to 1100 h values (P less than 0.05), and levels 20 min post-ACTH injection at 1700 h were significantly greater in old than young or middle-aged rats at the same time (P less than 0.05). These results demonstrate that 1) there are no age-related changes in the diurnal secretion of corticosterone in Fischer 344 male rats; 2) there is a decline in the peak level of ACTH during the diurnal surge of old compared to young animals; and 3) adrenal sensitivity to ACTH at 1700 h is greater in old compared to young or middle-aged rats. We hypothesize that the greater increase in adrenal sensitivity to ACTH is responsible for the maintenance of the corticosterone rhythm in the presence of diminished ACTH concentrations in older rats. PMID- 3032585 TI - The hepatic glucagon receptor: a comparative study of the regulatory and structural properties. AB - Guanine nucleotide and Mg2+ ion regulation of [125I-Tyr10]monoiodoglucagon ([125I]MIG) binding to liver plasma membranes from chicken, rat, and rabbit was studied. It was found that [125I]MIG binding to chicken liver membranes was increased by the addition of Mg2+ ion, while binding to rat and rabbit liver membranes was unaffected. In the chicken liver membranes, the Mg2+ ion induced high affinity binding which was sensitive to guanine nucleotides, while the low affinity binding in the absence of Mg2+ ion was not. Maximal effects of Mg2+ ion were observed at 1 mM. Glucagon binding to rat liver membrane receptors was GTP sensitive regardless of whether Mg2+ ion was added. Glucagon binding to rabbit liver membranes was insensitive to both Mg2+ ions and GTP. This lack of GTP effect was not due to degradation of GTP; no effect of the nonhydrolyzable analog guanyl-5'-yl-imidodiphosphate was observable. Glucagon stimulation of rabbit liver adenylyl cyclase, however, was dependent on GTP, as was the case with all of the other liver adenylyl cyclases studied here. The Kact of GTP for the rabbit liver system was very similar to that for rat liver membranes. The glucagon receptor was covalently labeled with [125I]MIG using p-hydroxysuccinimidyl azidobenzoate and analyzed by sodium dodecyl sulfate-gel electrophoresis. In all cases, a major labeled band at 63,000 daltons was observed. The levels of glucagon receptor and stimulatory (Ns) and inhibitory (Ni) regulatory proteins of adenylyl cyclase were measured. The highest levels of glucagon receptor were measured in rat liver membranes, while the levels in chicken and rabbit membranes were 30-40% lower. Rabbit liver membrane had the highest levels of Ns, while rat liver membranes had 2-fold lower and chick liver membrane 4-fold lower levels than rabbit liver membranes. The levels of Ni was similar in the three systems. Thus, the ratio of Ns to glucagon receptor was highest in the rabbit. In the rat, this ratio was 3-fold lower than that in the rabbit. In the chicken membranes, the ratio was about 60% of that in the rat. These data suggest that the observed differences in effects of GTP on hormone binding can be explained by alterations in the ratio of the receptor and Ns proteins among the various species. PMID- 3032586 TI - Identification of a corticotropin-releasing factor-binding protein in the plasma membrane of AtT-20 mouse pituitary tumor cells and its regulation by dexamethasone. AB - CRF stimulates the synthesis and secretion of proopiomelanocortin-derived peptides from AtT-20 mouse pituitary tumor cells. This study has shown that there is a specific binding site for CRF located on the plasma membrane of these cells. Both [125I]iodo-Tyr0CRF and noniodinated CRF (10(-11)-10(-7) M) stimulated, in a dose-dependent manner, the secretion of equimolar amounts of beta-endorphin-like immunoactivity from AtT-20 cells. Disuccinimidyl suberate, a cross-linking agent, was used to demonstrate specific binding of [125I]iodo-Tyr0CRF to plasma membranes from these cells. After cross-linking [125I] iodo-Tyr0CRF, the membrane proteins were solubilized with sodium dodecyl sulfate and electrophoresed on a 10% polyacrylamide gel. A single radioactively labeled band, corresponding to a mol wt of 66,000, was identified by autoradiography. [125I]Iodo-Tyr0CRF binding to these membranes was inhibited by 10(-7) M unlabeled CRF or an equimolar concentration of the CRF analog sauvagine. Similar concentrations (10(-7) M) of TRH, GnRH, insulin, [Arg8]vasopressin, somatostatin, and ACTH did not inhibit [125I]iodo-Tyr0CRF binding to the plasma membranes. Incubation of AtT-20 cells for 24 h in the presence of 10 nM dexamethasone reduced [125I]iodo-Tyr0CRF binding by 80% compared to that in untreated cells. Dexamethasone also inhibited the CRF-stimulated beta-endorphin-like immunoactivity secretory response. These data indicate that binding of CRF to a specific membrane protein is an integral component in the stimulation of AtT-20 cells by CRF. PMID- 3032587 TI - Adrenocorticotropin and the time of day both influence the amount and activity of 3-hydroxy-3-methylglutaryl coenzyme-A reductase in the hamster adrenal. AB - We studied the effect of ACTH, alone or in combination with cycloheximide, on hamster adrenal 3-hydroxy-3-methylglutaryl coenzyme-A (HMG-CoA) reductase protein content (mass) and activity. Immunoblotting was performed on both homogenate and microsomal preparations, using a rabbit antirat liver reductase antibody and 125I labeled protein A. A dose-dependent increase in HMG-CoA reductase protein content and activity was produced by ACTH. A time-course study revealed a latency of 60 min, followed by a significant increase at 120 and 180 min, in this response. At 180 min, microsomal reductase activity was significantly increased 4.7-fold, whereas the reductase protein content of microsomes and homogenate preparations was enhanced 4.4- and 3.1-fold, respectively. We calculated that only 40-50% of the reductase protein content was microsomal, indicating that more than one pool of the enzyme is present in adrenals. The coadministration of cycloheximide with ACTH not only prevented the hormonal effect on the protein content and activity of the reductase, but also produced, 180 min posttreatment, 58-69% and 65-86% decreases in reductase protein and activity, respectively. Also, both reductase activity and protein content fluctuated in parallel during the day. In the presence of proteolytic enzyme inhibitors and iodoacetamide, a major band in the area of 102K mol wt was evidenced by immunoblotting. These results indicate that ACTH and other conditions that provoke a change in adrenal HMG-CoA reductase activity also induce, in parallel, a change in the reductase protein content. The basic unit of the reductase has an apparent mol wt of about 102K. PMID- 3032589 TI - Pituitary and adrenal responses to pulsatile ovine corticotropin-releasing factor administered to fetal sheep. AB - In fetal sheep a bolus injection of ovine CRF (oCRF) elevates plasma immunoreactive ACTH (IR-ACTH) during the last 5 weeks of gestation. However, the effects of long term administration of oCRF to fetal sheep have not been studied. We examined the effects of pulsatile administration of oCRF (1 microgram every 4 h) for 7 days on fetal pituitary and adrenal responses, as reflected by plasma concentrations of IR-ACTH and cortisol (F). In addition, we examined the effects of oCRF on cAMP accumulation by dispersed pituitary cells in vitro after treatment in vivo with either oCRF or saline. Pulsed oCRF (P-CRF) treatment resulted in a significant (P less than 0.05) increase in basal IR-ACTH and F concentrations on all days of treatment. However, the pituitary response (change in IR-ACTH in response to a pulse of oCRF) decreased, and the adrenal response (change in F in response to endogenously secreted ACTH) increased as treatment progressed. A significant inverse correlation (r = 0.962) between basal F and the IR-ACTH response to oCRF was seen over the 7 days of treatment. Although P-CRF treatment resulted in an increase in fetal adrenal weight, it did not lead to premature parturition. There was a dose-dependent accumulation of cAMP in response to oCRF in vitro by dispersed pituitary cells from both groups of fetuses. However, this response was significantly greater when the fetuses had been pretreated with oCRF in vivo than after saline treatment. We conclude that the P-CRF regimen employed in this study stimulates the fetal pituitary-adrenal axis and the ability of fetal pituitary cells to accumulate cAMP in response to further oCRF in vitro. The reduced plasma IR-ACTH response after continued P-CRF in vivo may be attributed to increasing negative feedback effects of elevated endogenous F. PMID- 3032588 TI - 2-Hydroxyestradiol modulates a facilitative action of catecholamines on porcine granulosa cells. AB - Recent studies have disclosed a novel intraovarian paracrine system whereby catecholestrogens [e.g. 2-hydroxyestradiol (2-OH-E2)], synthesized locally from estradiol (E2) can stimulate progesterone secretion by granulosa cells (GC). Since these studies suggested that effects of 2-OH-E2 were discrete from those of E2, the present studies were undertaken to determine if the effects of 2-OH-E2 could be mediated through or interact with the catecholamine response system of GC. First, the effects of 2-OH-E2 were compared with those of E2 and epinephrine (EPI) using undifferentiated porcine GC. After 4 days of treatment, saturating concentrations of EPI (1 microgram/ml), 2-OH-E2 (4 micrograms/ml), and E2 (1 microgram/ml) stimulated progesterone production per cell 2-, 9-, and 10-fold, respectively. Saturating doses of EPI plus 2-OH-E2 or EPI plus E2 caused further significant increases in progesterone production above the effects of any single treatment, and the effects of EPI plus 2-OH-E2 were significantly greater than that of EPI plus E2. Isoproterenol mimicked the effect of EPI. Neither propranolol (a beta-antagonist) nor phentolamine (an alpha-antagonist) blocked the effects of 2-OH-E2, suggesting that effects of 2-OH-E2 were not mediated through alpha- or beta-adrenergic receptors. Collectively, these data suggest that 2-OH-E2, beta-adrenergic agonists, and E2 use separate response systems. To further evaluate the interaction between catecholamines and 2-OH-E2, GC were treated with increasing doses of 2-OH-E2 with or without EPI or isoproterenol. EPI and isoproterenol caused a synergistic increase in progesterone production above that induced by all doses of 2-OH-E2 along (average 3.8 +/- 0.7- and 3.2 +/ 0.4-fold enhancement for EPI and isoproterenol, respectively). This synergistic effect was blocked with addition of propranolol, indicating a beta-adrenergic mediation for the catecholamine portion of this effect. Studies using the catechol-O-methyltransferase inhibitor, U-0521, and the O-methyl derivative of EPI, metanephrine, suggested that catechol-O-methyltransferase present in GC dramatically reduces the potency of EPI, but is not involved in the synergism between EPI and 2-OH-E2. In conclusion, 2-OH-E2 is a more efficacious stimulator of progesterone secretion than catecholamines and synergizes with beta-adrenergic agonists to further stimulate progesterone production by GC. The mechanism by which catecholamines and 2-OH-E2 interact within GC is unknown.4+owever, this catecholamine/2-OH-E2 interaction PMID- 3032590 TI - Hormonal regulation of phospholipid methyltransferase by 3',5'-cyclic adenosine monophosphate-dependent and independent mechanisms. AB - Treatment of isolated rat adipocytes with epinephrine or isoproterenol caused a time- and concentration-dependent increase in phospholipid methyltransferase (PLMT) activity that was blocked by propranolol and unaffected by phentolamine. Forskolin mimicked the stimulatory effect on PLMT, and insulin inhibited this effect. In both the absence and presence of insulin, there was a linear relationship between PLMT activity and lipolysis. PLMT activity was also increased in response to oxytocin, which does not activate adenylate cyclase in adipocytes and does not stimulate lipolysis. The effects of oxytocin were inhibited by insulin and were additive with those of isoproterenol on PLMT. These data support the hypothesis that in adipocytes, PLMT is activated by a cAMP dependent protein kinase and a cAMP-independent mechanism, both of which can be regulated independently, and both of which are sensitive to inhibition by insulin. PMID- 3032591 TI - Characterization and solubilization of vasoactive intestinal peptide receptors from rat lung membranes. AB - Receptors for vasoactive intestinal peptide (VIP) were characterized in rat lung membranes by binding and covalent cross-linking of [125I]VIP using ethylene glycolbis-(succinimidylsuccinate). Binding studies indicated the presence of two classes of binding sites for VIP in rat lung membranes: 0.28 +/- 0.11 pmol/mg protein high affinity receptors (Kd = 79.2 +/- 26.4 pM) and 3.3 +/- 0.9 pmol/mg protein lower affinity receptors (Kd = 4.8 +/- 2.1 nM). Furthermore, binding of [125I]VIP to rat lung receptors was inhibited by micromolar concentrations of GTP analogs, guanosine-5'-O-(3-thiotriphosphate) GTP gamma S), and guanylylimidodiphosphate, suggesting that VIP receptors in rat lung membranes were tightly coupled to the guanine nucleotide regulatory protein (Ns). Scatchard analysis of VIP binding in the presence of GTP gamma S revealed selective inhibition of binding to high affinity sites. A 58K band was specifically labeled when membranes covalently labeled with [125I]VIP were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions. The apparent size of this species was not altered when sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis was carried out in the absence of reducing agent. Unlabeled VIP inhibited the labeling, with an IC50 of about 1 nM. A related peptide, GH-releasing factor-(1-40)OH, exhibited a much lower binding affinity, and two unrelated peptides, insulin and atrial natriuretic factor, did not inhibit labeling of the 58K species, even at micromolar concentrations. Labeling of the 58K species was inhibited in a GTP gamma S-dependent manner, suggesting the involvement of this species in the coupling to Ns. These data collectively indicated that the 58K species was a VIP-binding unit of VIP receptors in rat lung membranes. Several nondenaturing detergents were tested for extraction of labeled receptors from the membrane; the best extraction was obtained using 1% n-octyl-beta-D-glucopyranoside. PMID- 3032592 TI - Direct stimulation of nucleoside triphosphatase activity in bovine luteal nuclear membranes by human chorionic gonadotropin. AB - Nuclear membranes of bovine corpora lutea contain nucleoside triphosphatase (NTPase), an enzyme involved in the nucleocytoplasmic transfer of mRNA. Upon addition to nuclear membranes, hCG stimulated this enzyme, but not Mg2+-ATPase or NADH cytochrome c reductase, in a dose-, time-, and temperature-dependent manner. Heat-denatured hCG, however, had no effect on NTPase activity. hCG antisera blocked hCG's ability to stimulate the NTPase activity. While human LH also stimulated luteal nuclear membrane NTPase activity, a variety of other hormones tested, including alpha- and beta-subunits of hCG and 1 and 10 mM cAMP, had no effect on the enzyme. hCG's effect on NTPase is tissue specific in that it had no effect on liver or kidney nuclear membrane NTPase activity. In conclusion, the present data demonstrate that hCG acts directly on the luteal cell nucleus, thus raising the possibility that internalized hCG might influence nuclear functions before it is eventually degraded in lysosomes of luteal cells. PMID- 3032593 TI - Solubilization and properties of oxytocin receptors from rat mammary gland. AB - Oxytocin (OT)-binding activity was extracted with the zwitterionic detergent 3 [(3-cholamidopropyl)dimethylammonio]2-hydroxy-1-propanesulfonate (CHAPSO) from rat involuted mammary glands with about 20% yield. The binding in detergent extracts was characterized and shown to be similar or identical to that of OT receptors on intact plasma membranes. Solubilized receptors had a high affinity site (Kd approximately 2 nM) and a lower affinity component, whereas the membrane receptor has only the high affinity site. Several synthetic OT analogs inhibited [3H]OT binding in the same rank order in both solubilized and intact membrane preparations. Both solubilized and membrane-associated receptor required Mn2+ for [3H]OT binding. The concentration of OT-binding sites in solubilized extracts of uterine myometrium from rats in late pregnancy was substantially greater in uteri from rats in labor than in that from rats 2 days before labor, as we have seen previously with receptors on intact membranes. The affinity of the solubilized myometrial receptor (Kd approximately 5 nM) was comparable to that of the membrane-associated receptor. Binding of [3H]OT to solubilized extracts of intestinal smooth muscle, which is not a target for OT, was negligible. Gel filtration analysis on columns of Sepharose 6B indicated that the solubilized [3H]OT-binding component from mammary gland was present in multiple mol wt forms, but the smallest major form eluted with an average apparent mol wt of about 40,000. These studies indicate that CHAPSO-solubilized binding sites for [3H]OT are the same as those in intact membranes and, therefore, are components of the OT receptor. PMID- 3032594 TI - 1,25-Dihydroxycholecalciferol receptor regulation in hormonally induced differentiation of mouse mammary gland in culture. AB - Mammary explants from pregnant mouse cultured in the presence of insulin, cortisol, and PRL express their differentiated function by producing milk proteins such as casein. Sucrose density gradient analysis of high salt extract of cultured explants showed the presence of 1,25-dihydroxycholecalciferol (1,25 (OH)2D3 specific binding activity sedimenting at 3.3 S. Specific 1,25-(OH)2D3 binding activity in mammary explants varied as a function of the hormonal milieu in culture. The highest activity was found in explants cultured in the presence of insulin, cortisol, and PRL, whereas only a marginal activity was present in explants cultured with insulin. The comparison of 1,25-(OH)2D3 binding activity in explants cultured with insulin and with the three hormone combination by Scatchard plot analysis indicated that the synergistic actions of insulin, cortisol, and PRL increased the number of 1,25-(OH)2D3 specific binding proteins as much as 6-fold; however, the affinity constant of the binding protein was relatively constant in the two systems. Time course studies showed that the increase in 1,25-(OH)2D3 binding activity was detectable as early as 12 h after the addition of cortisol and PRL to insulin-primed explants. The increase was inhibited by the presence of actinomycin D and cycloheximide, suggesting that the hormonal stimulation of 1,25-(OH)2D3 binding activity involves both transcriptional and translational process. These results indicate that insulin, cortisol, and PRL stimulate the specific 1,25-(OH)2D3 binding activity during the induction of mammary functional differentiation when milk protein synthesis takes place. PMID- 3032595 TI - Purification and partial characterization of two prolactin-like glycoprotein hormone complexes from the midpregnant mouse conceptus. AB - Two PRL-like glycoprotein hormone complexes were purified from the medium of cultured mouse conceptuses from day 10 of pregnancy: mouse placental lactogen-I (mPL-I) (29-32K), and mPL-I (36.5-42K). Sodium dodecyl sulfate-gel electrophoresis revealed that mPL-I (36.5-42K) is a complex of five proteins with mol wt of 36.5K, 37.5K, 39K, 40.5K, and 42K. Deglycosylation with peptide: N glycosidase F or trifluoromethanesulfonic acid produced a single 29K protein. mPL I (36.5-42K) was also sensitive to neuraminidase, but not to endo-beta-N acetylglucosaminidase H or bacterial alkaline phosphatase. The production of intermediates from partial digestion of mPL-I (36.5-42K) with endo-beta-N acetylglucosaminidase F indicated the presence of multiple glycosylation sites. mPL-I (29-32K) is a complex of three proteins with mol wt of 29K, 30.5K, and 32K. Treatment with peptide:N-glycosidase F or trifluoromethanesulfonic acid reduced the mol wt of the 30.5K and 32K bands to 28K. The 30.5K band was sensitive to endo-beta-N-acetylglucosaminidase H and endo-beta-N-acetylglucosaminidase F, but the 32K band was not. Neither band was sensitive to neuraminidase or bacterial alkaline phosphatase. The 29K band was resistant to all chemical and enzymatic treatments and is probably not glycosylated or phosphorylated. In the nonreduced state, neither form of mPL-I showed an increase in mobility over that of its reduced counterpart on sodium dodecyl sulfate-gel electrophoresis, indicating that neither form of mPL-I contains the large disulfide loop common to hormones of the PRL family. After iodination, all component proteins of both forms of mPL I were found to bind to day 17 pregnant mouse liver membranes and were displaceable by excess mPL-II. In a radioreceptor assay, 125I-labeled mPL-I (36.5 42K) was displaced by mPRL or mPL-II, but not by mGH. An antiserum to both forms of mPL-I was generated, and a RIA employing mPL-I (36.5-42K) as the standard and radioligand was developed. Dilutions of day 10 pregnant maternal mouse serum and placental homogenate and a partially purified fraction of mPL-I (29-32K) produced displacement curves parallel to that of mPL-I (36.5-42K) standard curve. Five micrograms of mPRL, mPL-II, or mGH or 10 microliter day 17 pregnant or male mouse serum did not displace the radioligand from the antibody. mPL-I (36.5-42K) was lactogenic, but it did not possess LH-like bioactivity. PMID- 3032596 TI - Steroidogenesis in adrenal tumor cells: influence of cell shape. AB - Y-1 adrenal tumor cells were grown on plastic, or plastic treated with poly(2 hydroxyethyl methacrylate) (polyHEMA) to produce concentration-dependent rounding (10(-5)-3 X 10(-4) M) of the cells or on plastic treated with poly-D-lysine (polylysine) to produce flat cells, in order to determine whether or not cell shape is correlated with steroid synthesis. The degree of rounding of cells was measured by determining mean cell height and longest cell diameter. Three measurements of steroid production were made: production of 20 alpha dihydroprogesterone, transport of cholesterol to the inner mitochondrial membrane, and production of pregnenolone by isolated mitochondria. Cells grown on poly(HEMA) showed increase in mean cell height, decrease in longest diameter (i.e. rounding), and increase in all three measurements of steroidogenesis. In the case of synthesis of 20 alpha-dihydroprogesterone, the response was dependent on the concentration of poly(HEMA), being greater with higher concentrations (up to 10(-4) M), of this agent. Moreover the degree of rounding (cell height) was correlated with production of 20 alpha-dihydroprogesterone at three concentrations of poly(HEMA) (r = 0.93. ACTH at a supramaximal concentration produced increases in all of these responses to the poly(HEMA) surface. Polylysine produced flatter cells (lower mean height and greater longest diameter) than plastic and also inhibited all three steroidogenic responses to ACTH. (Bu)2cAMP exerted the same effects as ACTH. Growing cells on poly(HEMA) or polylysine did not affect production of cyclic AMP by the cells. Addition of poly(HEMA) or polylysine to the medium in which the cells were incubated, at the same concentrations as those used for influencing cell shape, was without effect on steroid synthesis or the response to ACTH. Cells grown on poly(HEMA) show decreased incorporation of [3H] thymidine into DNA. It is concluded that cell shape influences the delivery of cholesterol to inner mitochondrial membrane and in this way, increases the production of steroids by Y-1 cells and that the effects of poly(HEMA) on cell shape, cholesterol transport, and synthesis of DNA may involve microfilaments. PMID- 3032597 TI - Thyrotropin stimulates glucose transport in cultured rat thyroid cells. AB - Glucose transport by FRTL-5 cells, a rat thyroid cell line, was found to be TSH dependent. The effect of TSH on the uptake of 2-deoxy-D-glucose, a nonmetabolizable glucose analogue, was prompt, being 200% over basal value after 10 min and maximal after 12 h (600-700% increase). The TSH effect was dose dependent, with half-maximum stimulation at 10 microU TSH/ml, and maximum stimulation at 1 mU TSH/ml. TSH enhanced also the uptake of 3-O-methyl-D-glucose by FRTL-5 cells. The TSH activation of glucose transport had the following characteristics: it was mimicked by (Bu)2-cAMP (1 mM) and by agents that increase cAMP levels in thyroid cells, such as forskolin (10 microM) and cholera toxin (50 micrograms/ml); it involved the facilitated glucose transport system in that it was inhibited in a dose-related manner by both cytochalasin B and phloretin; it showed a glucose stereochemical sensitivity, being affected by D-glucose and 3-O methyl-glucose, and not by L-glucose; it was characterized by an increase in the maximum velocity (Vmax) of glucose uptake (from 15.3 to 66.0 fmol/min X micrograms DNA) without change in the Michaelis-Menten constant (Km) (5.3 mM); the effect on the Vmax was due to an increase in the number of surface glucose transporters as indicated by the enhancement of the D-glucose-sensitive fraction of [3H]cytochalasin B binding sites that in thyroid plasma membranes of cells exposed to TSH for 2 and 8 h, increased from 5.0 (basal value) to 10.4 and 23.1 pmol/mg protein, respectively. These data indicate that in FRTL-5 cells TSH stimulates the glucose transport system by an enhancement of the number of functional glucose transporters in the thyroid plasma membrane. PMID- 3032599 TI - Quantitation of silica-induced type II cell hyperplasia by using alkaline phosphatase histochemistry in glycol methacrylate embedded lung. AB - A light microscopic technique based on alkaline phosphatase histochemistry was developed to specifically quantitate Type II cells in the intact rat lung. Lungs were fixed in 4% neutral-buffered formalin containing 0.25 M sucrose and embedded in glycol methacrylate. Two micron thick sections were mounted on glass microscope slides. Alkaline phosphatase activity was localized by using naphthol AS-BI phosphate as substrate in 0.125 M 2-amino-2-methyl-1-propanol buffer containing 0.625 mM MgCl2 (pH 8.9). Sections were counterstained with Harris hematoxylin. Type II cells were the only cell type in the alveolar region containing alkaline phosphatase activity, an observation that was confirmed by using electron microscopic histochemistry. By combining the alkaline phosphatase staining technique with standard morphometric procedures, the proliferative response to a single intratracheal dose of 10 mg silica was followed as a function of time. Type II cells were significantly increased at all time points examined. Twenty eight days following silica, Type II cells had increased to (252 +/- 16) X 10(6) cells per set of lungs compared to a control value of (141 +/- 32) X 16(6) cells. The method presented is a simple and rapid technique for examining Type II cell population kinetics. PMID- 3032598 TI - An EPR study of the interactions between heme and flavin in yeast flavocytochrome b2. AB - E.P.R. experiments and spin-lattice relaxation time measurements have been performed on Flavocytochrome b2 in the range 10 K to 100 K, to obtain information on the distance between the two prosthetic groups of the protein, flavin and heme. We have used the stabilization effect of pyruvate on the semiquinone form of the flavin, to compare the E.P.R. spectral shape and the relaxation properties of the radical when the heme is either in the ferrous form or in the ferric form. When the heme is ferric, no significant increase of the line broadening or enhancement of the relaxation rate of the radical can be detected in the range 10 K to 100 K. From these results, a minimum intercentre distance of 18 to 20 A can be estimated. PMID- 3032600 TI - Physiological stress induced by sublethal concentrations of phenolic compounds in Notopterus notopterus: measurement of hydrolytic enzymes. AB - The studies on the effects of phenol (P), 2,4-dinitrophenol (DNP), pentachlorophenol (PCP) individually and in three combinations, viz., (PCP + DNP)/P (highly antagonistic), (DNP + P)/PCP (additive), and (P + DNP)/PCP (highly synergistic), at three sublethal levels (1/10, 1/15, and 1/20 of the 96-hr LC50) on the activity of hydrolytic enzymes (acid, alkaline, and glucose-6-phosphatases and 5-nucleotidase) in the brain and kidney of a fresh-water teleost, Notopterus notopterus revealed inhibition of enzyme activity after 15 and 30 days of exposure. Maximum inhibition (-89.32%) was observed in acid phosphatase activity in brain at 1/10 concentration of (P + DNP)/PCP combination after 30 days, but minimum (-3.64%) in activity of glucose-6-phosphatase in kidney at 1/20 of (PCP + DNP)/P combination after 15 days of exposure. However, biphasic effects of P, DNP, and (PCP + DNP)/P combination were observed in kidney, i.e., inhibition of enzymes activity at higher concentrations and stimulation at lower concentrations after both time intervals. PMID- 3032601 TI - The lung biological activity of American attapulgite. AB - Attapulgite is a fibrous mineral industrially consumed at the rate of over a million tons per year but the biological activity of the material is not fully known. To evaluate the in vivo toxicity of the fibrous material, we exposed the tracheal lobe of 16 sheep to a single exposure of either 100 ml saline, 100 mg UICC asbestos fibers in 100 ml saline, 100 mg short asbestos fibers in 100 ml saline, or 100 mg attapulgite in 100 ml saline. The animals were studied by bronchoalveolar lavage (BAL) at Days 2, 12, 24, 40, and 60 and by autopsy at Day 60. In the saline-exposed sheep, BAL and lung histology did not change. In the UICC asbestos-exposed animals, we reproduced the BAL changes previously reported (R. Begin, M. Rola-Pleszczynski, S. Masse, Y. Berthiaume, and G. Drapeau, 1985, Environ. Res. 36, 389-404). In the short asbestos-exposed sheep, there were no significant BAL changes. In the attapulgite sheep, we found significant and sustained increases in total BAL cells (X2, P less than 0.05), macrophages (X2, P less than 0.05), neutrophils (X5, P less than 0.01), fibronectin (X2-3, P less than 0.05), lactate dehydrogenase (X2, P less than 0.05), beta-glucuronidase (X2, P less than 0.05), but BAL cellularity returned to control levels by Day 60 whereas in the UICC asbestos-exposed sheep, it remained significantly above control. Lung histology demonstrated the characteristic peribronchiolar fibrosing alveolitis in the UICC asbestos-exposed sheep, whereas macrophagic alveolitis with minimal airway distortion was seen in the short asbestos-exposed sheep and in all of the attapulgite-exposed sheep but three which had typical peribronchiolar alveolitis quite similar to that observed in UICC-exposed sheep, but of lower intensity. In conclusion, the study documents that attapulgite is not an inert material for lung tissue; it does produce a macrophagic and neutrophilic alveolitis in the early inflammatory phase of lung reaction with peribronchiolar involvement in some sheep. Whether or not this initial attapulgite induced alveolitis leads to lung fibrosis remains to be evaluated in a longer follow-up study of attapulgite-exposed animals. PMID- 3032602 TI - Some biological properties of respirable iron ore dust. AB - The respirable fraction of an ore dust from the North-West of Western Australia was tested for biological properties by inhalation and intrapleural implantation trials using rats and mice. Pulmonary histology indicated significant levels of interstitial pneumonia occasionally associated with bronchopneumonia, bronchiectasis, emphysema, and lung collapse over that found in age-matched control animals. While there was a significant increase of the incidence of tumors in general in WAG inbred rats up to 2 years following dust exposure, this did not persist into old age. No mesotheliomas were induced by any treatments associated with iron ore dust, although the rats were shown to be susceptible to crocidolite asbestos-induced mesothelioma. In the mouse models, tumors which are normally seen only in aged animals were induced with a significant number of bronchial adenomas being recorded following intrapleural implantation of dust into inbred BALB/c mice. Leukemia/lymphoma associated with murine leukemia virus was increased following dust inhalation by inbred C57BL mice. PMID- 3032603 TI - The prophylactic use of polyvinylpyridine-N-oxide (PVNO) in baboons exposed to quartz dust. AB - Twelve baboons were exposed to a quartz dust cloud. Four of these were also given polyvinylpyridine-N-oxide (PVNO) by aerosol and four received PVNO by aerosol and injection. A prophylactic effect was demonstrated during the course of treatment, but when treatment stopped the silicosis progressed to the same degree of severity as in the untreated animals. PMID- 3032604 TI - Activation of latent collagenase from polymorphonuclear leukocytes by oxygen radicals. AB - Polymorphonuclear leukocytes (PMN) accumulating at inflammatory sites have the potential to degrade collagen by releasing the metalloproteinase collagenase (EC 3.4.24.7), which is stored within the specific granules of these cells in a latent, inactive, form. In order to elucidate the activation mechanism the latent enzyme (molecular weight 91,000) was purified from human PMN and incubated with the oxygen radical-generating system of xanthine oxidase (EC 1.1.3.22) and hypoxanthine. This coincubation resulted in the activation of the latent enzyme as assessed by the collagenolytic attack on human and bovine cartilaginous tissue. Two parameters for collagenolysis were used: loss of hydroxyproline containing fragments, and mechanical measurements reflecting the stability of tissue specimens. Superoxide dismutase (EC 1.15.1.1) as well as catalase (EC 1.11.1.6) were capable of inhibiting the activation of latent PMN collagenase by the oxygen radical-generating system. The results indicate the hydroxyl radical to be the final oxidant responsible for the activation of latent PMN collagenase. Thus a new activation mechanism of latent collagenase is presented in this paper and discussed together with the potential relevance in pathophysiologic states of acute and chronic inflammation. PMID- 3032605 TI - Involvement of iron and iron-catalyzed free radical production in ethanol metabolism and toxicity. AB - Lipoperoxidation, a degradative process of membranous polyunsaturated fatty acids, has been suggested to represent an important mechanism in the pathogenesis of ethanol toxicity on the liver and possibly also on the brain. Catalysis by transition metals, especially iron, is involved in the biosynthesis of free radicals contributing to lipid peroxidation. Although the exact nature of the redox-active iron implicated in this catalysis is still unknown, it has been well established that lipid peroxidation can be prevented in vitro by iron chelators such as desferrioxamine. Deprivation of redox-active iron through desferrioxamine inhibits by about 50% the microsomal oxidation of ethanol in vitro and reduces very significantly in vivo the overall ethanol elimination rate in rats. Administration of desferrioxamine together with ethanol also reduces the ethanol induced disturbances in the antioxidant defense mechanisms of the hepatocyte. It also reduces in mice both the severity of physical dependence on ethanol and lethality following the acute administration of a narcotic dose of ethanol. Chronic overloading of rats with iron results, on the opposite, in an increased rate of ethanol elimination, although alcohol dehydrogenase and catalase activities are reduced and cytochrome P-450 depleted in the liver of such iron overloaded animals. The magnitude of the ethanol-induced increase in lipid peroxidation and decrease in the major membranous antioxidant, alpha-tocopherol, is exacerbated in iron-overloaded rats. Several disturbances of iron metabolism have been reported in human alcoholics. Their contribution to ethanol toxicity appears very likely in the case of hepatic siderosis associated with alcohol abuse. Ethanol could however disturb iron metabolism even in the absence of gross abnormalities of the total iron stores. It is suggested that ethanol intoxication could increase cellular redox-active iron, thus contributing to an enhanced steady-state concentration of reactive-free radicals. This oxidative stress would lead to lipoperoxidative damage and cellular injury. PMID- 3032606 TI - Effects of ethanol on the activity of brain enzymes. AB - Ethanol alters, in a selective manner, the activity of several membrane-bound enzymes in the central nervous system (CNS) which are important for neuronal transmission of information. Ethanol inhibits Na+/K+-transporting ATPase activity, while adenylate cyclase (AC) activity is stimulated by ethanol added in vitro. Ethanol's effects on AC activity are mediated primarily via effects on proteins that regulate AC activity. Ethanol has selective effects on monoamine oxidase activity, in that the B form of the enzyme is more sensitive to inhibition by ethanol added in vitro. The selective effects of ethanol on different membrane-bound CNS enzymes may result from differing membrane lipid microenvironments of the enzymes, or from differences in the enzyme proteins per se. PMID- 3032607 TI - Comparative in vitro activity of LY146032 (daptomycin), a new lipopeptide antimicrobial. AB - The in vitro activity of LY146032, a new cyclic lipopeptide antibiotic, was compared with those of vancomycin, teicoplanin, and either oxacillin or ampicillin by determining agar dilution MIC values for 304 clinical gram-positive isolates. LY146032 had superior in vitro activity against oxacillin-resistant staphylococci when compared to vancomycin or teicoplanin. Against oxacillin sensitive staphylococci, group JK-diphtheroids, streptococci, Listeria monocytogenes and Clostridium difficile, LY146032 was equally or less active than vancomycin, teicoplanin, or the penicillins tested. When tested by macrobroth dilution MIC/MBC, LY146032 showed good bactericidal activity against all organisms with the exception of Clostridium difficile. PMID- 3032608 TI - Enzyme profile of Haemophilus ducreyi strains isolated on different continents. AB - Two hundred strains of Haemophilus ducreyi, isolated in different parts of the world, were investigated using the API-ZYM system, which included 95 different substrates. All strains produced aminopeptidase against beta-naphthylamide derivatives of L-lysine, glycine, L-arginine, L-alanine, D-L-methionine, glycyl glycine, glycyl-L-alanine and L-leucine. All strains also produced alkaline and acid phosphatase and phosphohydrolase. Nearly all strains showed esterase activity against butyrate, valerate, caproate and caprylate, but the reactions were very weak. No glycosidase activity could be detected. Of the 47 aminopeptidase tests showing variable reactions, only the results for S-benzyl-L cystine, L-ornithine, L-alanyl-L-phenyl-alanyl-L-proline, L-hystidyl-L-leucyl-L histidine and L-histidyl-L-serine arylamidase obtained on strains from Asia, Africa and Europe were significantly different (p less than 0.05). On the basis of test results for L-ornithine arylamidase and L-alanyl-L-phenyl-alanyl-L proline arylamidase, the distribution of three biovars found among the isolates of the different continents was significantly different (p less than 0.0001), whereas African strains isolated in Kenya and South Africa yielded the same enzymatic pattern. Thus, these enzymes may constitute a marker system for the epidemiological study of Haemophilus ducreyi. PMID- 3032609 TI - A luminescence Western blot with enhanced sensitivity for antibodies to human immunodeficiency virus. AB - A luminescence assay was adapted for detection of peroxidase-labelled secondary antibodies, which react specifically with antibodies to human immunodeficiency virus on Western blots. This method is about one hundred times more sensitive than either commercial enzyme-linked immunosorbent assay or the chromogenic peroxidase assay on Western blots and detects seroconversion earlier than any other technique currently available. PMID- 3032610 TI - Reliability of the colistin disk test in identification of Serratia marcescens and Serratia liquefaciens. AB - Resistance to polymycin B or E (colistin) in the disk test, which is used as a means of identification, was tested in 43 strains of Serratia marcescens and 26 of Serratia liquefaciens. While all strains had MIC values greater than 32 mg/l for colistin, false susceptibility to both polymyxins was encountered in the 24 h disk test in 9% of Serratia marcescens and 12-96% of Serratia liquefaciens strains, depending on the kind of polymyxin, temperature and duration of incubation. Using colistin disks and incubation at 25 degrees C for 48 h, the percentage of false susceptible results could be minimized. PMID- 3032611 TI - In vitro activity of LY146032 (daptomycin), a new peptolide. AB - The in vitro activity of LY146032, a new peptolide antibiotic, was compared with those of vancomycin, teicoplanin, imipenem, amoxicillin and erythromycin. LY146032 inhibited 90% of Staphylococcus aureus and Staphylococcus epidermidis, including methicillin-resistant isolates at less than or equal to 1 microgram/ml. Its activity was comparable to those of vancomycin and teicoplanin. MIC90s for the beta-hemolytic streptococci varied from 0.25 microgram/ml for group B streptococci to 4 micrograms/ml for some group C and F streptococci. MICs for Streptococcus faecalis were in the range of 0.5 to 8 micrograms/ml, and the MIC90 4 micrograms/ml, compared to 4 micrograms/ml for vancomycin and 1 microgram/ml for teicoplanin. For some viridans streptococci the MICs were 4 micrograms/ml, whereas Streptococcus pneumoniae were inhibited by 0.5 microgram/ml. Corynebacterium JK species were inhibited by 0.5 microgram/ml, similar to vancomycin, and Listeria monocytogenes by 4 micrograms/ml. Neisseria species, Haemophilus species and enteric species were not inhibited. Most MBCs were within two-fold of the respective MICs. After 14 days passage in sub-inhibitory concentrations of LY146032, Staphylococcus aureus, Staphylococcus epidermidis and Streptococcus faecalis showed minimal increase in MICs. The activity of LY146032 was increased by adding Ca2+ and was reduced in an anaerobic environment. Overall, LY146032 is an extremely interesting new agent that inhibits gram positive species. PMID- 3032612 TI - In vitro activity of LY146032 (daptomycin) against selected aerobic bacteria. AB - The in vitro activity of the new lipopeptide antibiotic LY146032 was generally four-fold greater (MIC 90 less than or equal to 0.5 microgram/ml) than that of vancomycin against methicillin-susceptible or methicillin-resistant Staphylococcus aureus and coagulase-negative species of Staphylococcus. Enterococci, Streptococcus bovis, group B and viridans streptococci, and Corynebacterium group J-K isolates were inhibited by less than or equal to 4 micrograms/ml of LY146032, which represented activity equivalent to or greater than that of vancomycin. Unlike vancomycin, LY146032 was bactericidal for Enterococcus faecalis, Enterococcus faecium and Listeria monocytogenes. Due to its bactericidal properties LY146032 appeared to represent an improvement over vancomycin and teicoplanin. PMID- 3032613 TI - Comparative in vitro activity of LY146032 (daptomycin) against gram-positive cocci. AB - The in vitro activity of LY146032 was compared with those of seven other antimicrobial agents against gram-positive cocci. MICs of LY146032 were lowest for Streptococcus pneumoniae and methicillin-susceptible Staphylococcus epidermidis (0.25 mg/l). For methicillin-resistant Staphylococcus epidermidis and Staphylococcus aureus the MICs were 1 mg/l, for Enterococcus faecalis 2 mg/l and for Enterococcus faecium 4 mg/l. The activity of LY146032 was in general higher than that of vancomycin. Time-kill studies showed LY146032 had higher bactericidal activity than vancomycin against a methicillin-resistant Staphylococcus aureus strain, and bactericidal activity against Enterococcus faecalis and Enterococcus faecium. PMID- 3032614 TI - Cytochrome c oxidase is a three-copper, two-heme-A protein. AB - Metal contents of preparations of procaryotic (Paracoccus denitrificans) and eucaryotic (beef heart) cytochrome c oxidases have been determined by inductively coupled plasma atomic emission spectroscopy and shown to be stoichiometrically related to the protein contents. The results show that oxidases which possess subunits I and II have three copper atoms besides hemes a and a3 (Paracoccus denitrificans, Cu: 2.97 +/- 0.08 and Fe: 2.09 +/- 0.10; bovine heart, Cu: 2.83 +/ 0.07 and Fe: 1.94 +/- 0.12). Together with data reported for the c1 aa3 oxidase from Thermus thermophilus, the following conclusions can be drawn. Subunit I binds two copper atoms and both hemes a and a3 and thus is the universal terminal oxidase of this spectral type. Subunit II binds one copper and functions as an electron conductor. The mitochondrial respiratory complex IV binds, in addition to three copper and two hemes a, stoichiometric amounts of magnesium and zinc (bovine heart Mg: 0.98 +/- 0.05 and Zn: 1.01 +/- 0.04). PMID- 3032615 TI - Regulation of the cholesterol ester cycle of cultured Leydig tumor cells. AB - The MA-10 Leydig tumor cells take up low-density lipoprotein (LDL) from the medium and store the LDL-derived cholesterol as cholesterol esters that can be subsequently mobilized and used for steroid hormone synthesis. The present studies investigate the mechanisms by which cAMP acutely regulates the cellular content of cholesterol esters. In the absence of cholesterol utilization for steroidogenesis, cAMP stimulates cholesterol ester hydrolysis and ester resynthesis proportionally. The augmentation of ester hydrolysis by cAMP is completely matched by increased activity of the acyl-coenzyme-A:cholesterol acyltransferase and thus does not regulate cellular cholesterol ester concentration per se. The more important action of cAMP is to interrupt the cycle of hydrolysis and ester resynthesis by decreasing cholesterol re-esterification. In cells actively synthesizing steroid hormones, cholesterol reesterification is decreased by 82%. The decrease in cholesterol re-esterification occurs because cAMP directs cholesterol normally destined for re-esterification into steroid synthesis; simply blocking the utilization of cholesterol for steroidogenesis completely prevents net cholesterol ester hydrolysis and increases the cellular rate of cholesterol esterification. PMID- 3032617 TI - Differentiation of the drug-binding sites of calmodulin. AB - Calmodulin contains several binding sites for hydrophobic compounds. The apparent specificity of various 'calmodulin antagonists' for these sites was investigated. The Ki values for the inhibition of calmodulin-activated cyclic-nucleotide phosphodiesterase and myosin light-chain kinase was determined. In addition, the Kd values of the same compounds for binding to calmodulin were measured. The compounds could be separated into four groups. Group I and II compounds inhibited competitively the activation of the phosphodiesterase and myosin light-chain kinase by calmodulin. Group I compounds inhibited the activation of the phosphodiesterase and myosin light-chain kinase at identical concentrations. In contrast, group II compounds inhibited the activation of the phosphodiesterase at 5-10-fold lower concentrations than that of myosin light-chain kinase. Group III compounds inhibited the activation of these enzymes by an uncompetitive mechanism. Group IV compounds inhibited the activation of the phosphodiesterase with Ki values above 10 microM and did not affect the activation of myosin light chain kinase. Binding of [3H]bepridil to calmodulin under equilibrium conditions yielded one high-affinity site (apparent Kd 0.4 microM) and four low affinity sites (apparent Kd 44 microM). Group I compounds interfered with the binding of bepridil to the high and low-affinity sites in a competitive manner. Group II compounds interfered in a non-competitive manner with the high-affinity site and apparently competed only with one of the low-affinity sites. Group III compounds did not compete with any of the bepridil-binding sites. Nimodipine, a group III compound, bound to one site on calmodulin with a Kd value of 1.1 microM. Other dihydropyridines competed with [3H]nimodipine for this site. The group I and II compounds, trifluoperazine and prenylamine, did not affect the binding of [3H]nimodipine. These data show that 'calmodulin antagonists' can be differentiated into at least three distinct groups. Kinetic and binding data suggest that the three groups bind to at least three different sites on calmodulin. Selective occupation of these sites may inhibit specifically the activation of distinct enzymes. PMID- 3032616 TI - Changes in the concentration of cAMP, fructose 2,6-bisphosphate and related metabolites and enzymes in Saccharomyces cerevisiae during growth on glucose. AB - Changes in the concentration of several metabolites and enzymes related to carbohydrate metabolism were measured during the growth of Saccharomyces cerevisiae on a mineral medium containing glucose as the limiting nutrient. When about 50% of the original glucose was used the exponential phase ended and the culture entered a 'transition' phase before the complete exhaustion of glucose. In this transition phase several metabolic changes occurred. cAMP, that decreased along growth, reached a constant value of about 0.7 nmol/g dry weight. A pronounced drop in fructose-6-phosphate-2-kinase activity and in the concentration of fructose 2,6-bisphosphate and fructose 1,6-bisphosphate was observed accompanied by a less marked decrease in hexose monophosphates. Trehalase activity also dropped and reached a minimal value at the onset of the stationary phase when synthesis of trehalose began. Glycogen concentration and glycogen synthase activity increased sharply during the transition phase. Plasma membrane ATPase began to increase at the middle of the exponential phase and then, coincident with the glucose exhaustion, a 90% decrease in the measurable activity was observed. PMID- 3032618 TI - A comparative study on the phoE genes of three enterobacterial species. Implications for structure-function relationships in a pore-forming protein of the outer membrane. AB - The cloned phoE genes from Enterobacter cloacae and Klebsiella pneumoniae are normally expressed and regulated in Escherichia coli K-12, and their products are correctly assembled into the outer membrane. Differences between the three PhoE proteins were found with binding of two out of ten monoclonal antibodies directed against the cell-surface-exposed part and in pore characteristics, but not in phage receptor function. The DNA sequences of the E. cloacae and K. pneumoniae phoE genes were determined and used to predict the primary structures of the encoded proteins. In the upstream non-coding regions, which showed more variations among the three genes than the coding regions, conserved sequences were identified which might be involved in regulation of phoE gene expression. Comparison of the predicted PhoE primary structures revealed a high degree of homology, with 81% of the amino acid residues being identical in all three proteins. Four small variable regions were found where differences are the most pronounced, corresponding to regions which were previously predicted to be exposed at the cell surface. Implications of the sequence comparison for structure-function relationships in PhoE protein are discussed. PMID- 3032620 TI - Characteristics of the redox-linked proton ejection in beef-heart cytochrome c oxidase reconstituted in liposomes. AB - In this paper a study is presented of the characteristics of redox-linked proton ejection exhibited by isolated beef-heart cytochrome c oxidase incorporated in asolectin vesicles. The enzyme was 90% oriented 'right-side out' as in the mitochondrial membrane. The effects on the H+/e- stoichiometry of the modalities of activation of electron flow, the pH of the medium and its ionic composition were investigated. The results obtained show that, whilst ferrocytochrome c pulses of the aerobic oxidase vesicles at neutral pH and in the presence of saturating concentrations of valinomycin and K+ to ensure charge compensation produced H+/e- ratios around 1 (as has been shown previously), oxygen pulses of reduced anaerobic vesicles supplemented with cytochrome c, gave H+/e- ratios around 0.3. The H+/e- ratios exhibited, with both reductant and oxidant pulses, a marked pH dependence. Maximum values were observed at pH 7.0-7.7, which decreased to negligible values at acidic pH with apparent pKa of 6.7-6.3. Mg2+ and Ca2+ caused a marked depression of the H+/e- ratio, which in the presence of these cations and after a few ferrocytochrome pulses, became negligible. Analysis of cytochrome c oxidation showed that the modalities of activation of electron flow and divalent cations exerted profound effects on the kinetics of cytochrome c oxidation by oxidase vesicles. The observations presented seem to provide interesting clues for the nature and mechanism of redox-linked proton ejection in reconstituted cytochrome c oxidase. PMID- 3032619 TI - Iron transport and storage. PMID- 3032621 TI - The calcium channel antagonists receptor from rabbit skeletal muscle. Reconstitution after purification and subunit characterization. AB - The Ca2+ channel antagonists receptor from rabbit skeletal muscle was purified to homogeneity. Following reconstitution into phosphatidylcholine vesicles, binding experiments with (+)[3H]PN 200-110, (-)[3H]D888 and d-cis-[3H]diltiazem demonstrated that receptor sites for the three most common Ca2+ channel markers copurified with binding stoichiometries close to 1:1:1. Sodium dodecyl sulfate gel analysis of the purified receptor showed that it is composed of only one protein of Mr 170,000 under non-reducing conditions and of two polypeptides of Mr 140,000 and 32,000 under disulfide-reducing conditions. Iodination of the protein of Mr 170,000 and immunoblots experiments with antisera directed against the different components demonstrated that the Ca2+ channel antagonists receptor is a complex of Mr 170,000 composed of a polypeptide chain of Mr 140,000 associated to one polypeptide chain of Mr 32,000 by disulfide bridges. One of the problems concerning this subunit structure of the putative Ca2+ channel was the presence of smaller polypeptide chains of Mr 29,000 and 25,000. Peptide mapping of these polypeptide chains and analysis of their cross-reactivity with sera directed against the proteins of Mr 170,000 and 32,000 demonstrated that they were degradative products of the Mr 32,000 component. Both the large (140 kDa) and the small (32 kDa) component of the putative Ca2+ channel are heavily glycosylated. At least 20-22% of their mass were removed by enzymatic deglycosylation. Finally the possibility that both the 140-kDa and 32-kDa components originate from a single polypeptide chain of Mr 170,000 which is cleaved by proteolysis upon purification is discussed. PMID- 3032622 TI - 5,6-Dichloro-1-beta-O-ribofuranosylbenzimidazole induces DNA damage by interfering with DNA topoisomerase II. AB - We have examined DNA in cells treated with 5,6-dichloro-1-beta-O ribofuranosylbenzimidazole (DRB), an adenosine analogue. The results show that DRB induces an partial fragmentation of DNA when the cells are lysed in dilute alkali. Fragmentation of DNA does not occur in control cells, nor in cells pretreated with novobiocin or VP-16/VM-26. The data show that DRB interferes with DNA topoisomerase II. In agreement with this interpretation, crude nuclear extracts of DRB-treated cells result in reduced in vitro KC1/SDS precipitation of covalent protein-DNA complexes. Formation of covalent complexes is typical of topoisomerase-DNA interaction. PMID- 3032623 TI - Glucagon antagonists. Synthesis and inhibitory properties of Asp3-containing glucagon analogs. AB - In an effort to find analogs of glucagon that would bind to the glucagon receptor of the rat liver membrane but would not activate membrane-bound adenyl cyclase, several hybrid molecules were synthesized which contained sequences from both glucagon and secretin. [Asp3, Glu9]Glucagon and [Asp3, Glu9, Arg12]glucagon were inactive in the adenyl cyclase assay even at high concentrations but retained some binding affinity for the receptor. They were able to displace 125I-glucagon completely from its receptor and could completely inhibit the activation of adenyl cyclase by natural or synthetic glucagon. The inhibition index [I/A]50 was approximately 110 for both analogs. [Asp3]Glucagon, [Glu3]glucagon and [Asp3, Lys17, 18, Glu21]glucagon were weak partial agonists, while [Asp3, Glu21]glucagon was inactive and a poor inhibitor. The peptides were synthesized by solid-phase methods and purified to homogeneity by reverse-phase high-performance liquid chromatography on C18 silica columns. These are the first fully synthetic competitive glucagon antagonists to be reported. PMID- 3032624 TI - Structural organization of the bovine thyroglobulin gene and of its 5'-flanking region. AB - The structural organization of the bovine thyroglobulin gene has been investigated by a combination of Southern genomic blotting and direct analysis of cloned gene fragments isolated from a chromosomal DNA library. The entire locus is spread over more than 200,000 base pairs which makes it one of the largest eukaryotic genes studies to date. The coding information is scattered into at least 42 exons, 34 of which have been precisely identified. A different evolutionary origin of the 5' and 3' regions of the gene is supported by the highly different proportion of exonic material they contain (12% and 3%, respectively) and by the existence of sequence homology between the 3' region of thyroglobulin and acetylcholinesterase. Detailed sequence analysis of the 5' region of the gene and its flanking segment demonstrated that a significant homology exists between bovine and human thyroglobulin sequences, except for the presence within the ruminant promoter region of a 220-base-pair sequence belonging to the bovine monomer repeated family. PMID- 3032625 TI - Stereospecific assignments of side-chain protons and characterization of torsion angles in Eglin c. AB - Stereospecific assignments were obtained in the protein Eglin c for beta methylene protons of 9 out of the 24 residues with AMX symmetry, for three residues with long side chains and for the gamma-methyl groups of 8 out of the 11 valines. This was achieved by analyzing the cross-peak multiplet structures in two-dimensional correlated spectra with spectral simulations and peak fitting, and by quantitative measurements of intraresidue nuclear Overhauser enhancements. In addition to the stereospecific assignments, information on the torsion angles phi and chi 1 was obtained. To estimate inferences of internal motions on this analysis, the effect of uniform averaging within a certain range of the torsion angle chi 1 on cross-peak multiplet structures and on relative intraresidue nuclear Overhauser enhancement is discussed. PMID- 3032626 TI - Solubilization and characterization of guinea-pig pancreatic somatostatin receptors. AB - The solubilization of somatostatin receptors from guinea-pig pancreas by different non-denaturing detergents was investigated after stabilization of the receptors by prior binding of 125I-[Tyr11]somatostatin or its analogue 125I [Leu8,DTrp22,Tyr25]somatostatin 28, to pancreatic plasma membranes. The somatostatin-receptor complexes were solubilized in a high yield by Zwittergent 3 14 (3-[tetradecyldimethylammonio]-1-propanesulfonate), a zwitterionic detergent. Other detergents, digitonin, Triton X-100, Chaps (3 [cholamidopropyldimethylammonio]-1-propanesulfonate) and octyl beta-D glycopyranoside, achieved only partial solubilization. The recovery of receptor complexes was increased by glycerol. In order to characterize solubilized somatostatin-receptor complexes, membranes receptors were covalently labelled using N-5-azido-2-nitrobenzoyloxysuccinimide as cross-linking reagent before solubilization. Gel filtration chromatography analysis resulted in the identification of a major protein component of apparent Mr = 93,000 which interacted with the two radioligands. In addition, a similar component of Mr = 88,000 was characterized after analysis by SDS-PAGE of membrane receptors covalently cross-linked with 125I-[Leu8,DTrp22,Tyr25]somatostatin 28 by different heterobifunctional reagents: N-5-azido-2-nitrobenzoyloxysuccinimide, N hydroxysuccinimidyl 4-azidobenzoate, N-succinimidyl 6-(4'-azido-2' nitrophenylamino)hexanoate. Optimal cross-linking results were obtained with N-5 azido-2-nitrobenzoyloxysuccinimide. The solubilized somatostatin-receptor complex was adsorbed to wheat-germ agglutinin-agarose column and eluted by specific sugars. We concluded that the guinea-pig pancreatic somatostatin receptor in the membrane and in the non-denaturing detergent solution behaves as a protein monomer of apparent Mr approximately 85,000-90,000. The somatostatin receptor is a glycoprotein which contains complex-type carbohydrate chains. PMID- 3032627 TI - Control of uncoupling protein in brown-fat mitochondria by purine nucleotides. Chemical modification by diazobenzenesulfonate. AB - The uncoupling protein (UP) of isolated brown adipose tissue mitochondria was studied with respect to the mechanism of control of UP function by purine nucleotides. Passive transport of H+ and Cl- was followed simultaneously in a KCl medium. With both GDP and ATP a higher sensitivity of Cl- transport (apparent Ki = 2.2 microM and 4.7 microM respectively) than of H+ transport (apparent Ki = 7.7 microM and 34 microM respectively) was observed. Chemical modification of isolated mitochondria by diazobenzenesulfonate (DABS) up to 75 mumol/mg protein did not affect the transport, its ionic selectivity and regulation by endogenous free fatty acids. In contrast, the sensitivity to purine nucleotides of both H+ and Cl- translocation was decreased (apparent Ki increased 71 and 47 times respectively). DABS decreased the affinity of [3H]GDP for the specific nucleotide binding site on mitochondria (Kd increased from 2.7 microM to 13 microM) and depressed, to a smaller extent, the GDP-binding capacity. Correlation between occupancy of the specific nucleotide-binding site by GDP and inhibition of transport yielded a linear relationship for Cl- transport in control mitochondria. For H+ transport in the control, and for both H+ and Cl- transports in DABS-treated mitochondria, a biphasic correlation was obtained. The results show that different structural parts of UP are involved in transport and its control by the regulatory ligands and that, in addition to binding of purine nucleotides to UP, the inhibition of ion transport by purine nucleotides depends on an intrinsic factor modulating the inhibitory effect. PMID- 3032628 TI - Characterization and regulation of somatomedin-C/insulin-like growth factor I (Sm C/IGF-I) receptors on cultured pig Leydig cells. Effects of Sm-C/IGF-I on luteotropin receptors and steroidogenesis. AB - We have characterized somatomedin-C/insulin-like growth factor I (Sm-C/IGF-I) receptors in cultured pig Leydig cells by binding and cross-linking affinity experiments. At equilibrium the dissociation constant and the number of binding sites per cell were 1.8 +/- 0.2 X 10(-9) M and 12,200 +/- 3200 respectively. Under reducing conditions, disuccinimidyl suberate cross-linked 125I-Sm-C to one receptor complex with apparent Mr = 120,000. Continuous treatment of Leydig cells with increasing concentrations of human chorionic gonadotropin (hCG) for 48 h resulted in a dose-dependent (ED50 0.05 nM) increment in IGF type I receptors (2.5-3-fold increase). Conversely, treatment of Leydig cells for 48 h with increasing concentrations of Sm-C/IGF-I produced a 3-4-fold increase in hCG receptors. This effect was dose-dependent (ED50 = 7 ng/ml). Sm-C/IGF-I treatment also enhanced Leydig cell responsiveness to hCG for both cAMP (6-fold) and testosterone (8-fold). Moreover the stimulatory effects of Sm-C/IGF-I on cells cultured either in the absence or presence of 5% human serum from an hypopituitary patient were inhibited by anti-(Sm-C/IGF-I) antibodies. Taken together these results not only show that Leydig cells must be considered as targets for Sm-C/IGF-I, but also lend support to the possibility that Sm-C/IGF-I plays a role in the regulation of Leydig cell function. Moreover, they suggest that Sm-C/IGF-I might be responsible for the delayed puberty observed in some patients with isolated growth hormone deficiency. PMID- 3032629 TI - The GTP pool in Bacillus brevis and its significance for sporulation. AB - The GTP pool of Bacillus brevis as well as that of other nucleotides is highly sensitive to all kinds of environmental changes like the cell transfer procedures or nutrient depletion of the cells. In growing cultures, as well as in cells transferred from rich to nitrogen-deficient medium, the nucleotide pool decreases significantly. This decrease is followed by the onset of sporulation only when cells are allowed to produce the peptide antibiotic tyrocidine or if tyrocidine is added to the culture. However, exogenous tyrocidine is active in triggering sporulation only when it is added within a short period of time immediately after shift down, that is when the nucleotide pool is observed to decrease. PMID- 3032630 TI - Ribonucleoside uptake and phosphorylation during fertilization and early development of the sea-urchin, Strongylocentrotus purpuratus. AB - Uptake of the four ribonucleosides normally present in RNA increases nearly 50 fold shortly after fertilization in eggs of the sea-urchin, Strongylocentrotus purpuratus. Uridine, adenosine and cytidine are phosphorylated (greater than 95%) to their mono-, di- and triphosphates immediately after transport into the fertilized egg. Although guanosine is transported to an extent equal to the other three ribonucleosides, less than 12% of its phosphorylated after transport. In vitro nucleoside and nucleotide kinase assays of unfertilized egg homogenates indicate that the uridine, adenosine and cytidine kinases as well as the uridylate, adenylate, cytidylate and guanylate kinases are present in the egg prior to fertilization. Substrate competition measurements indicate that adenosine phosphorylation is catalyzed by a monospecific enzyme, while uridine and cytidine phosphorylations are catalyzed by a common kinase. Guanosine kinase activity was not detectable in unfertilized egg homogenates. Between 3 h and 5 h after fertilization the phosphorylation of transported guanosine begins to increase as it enters the embryo. By 7 h after fertilization, more than 95% of the guanosine entering the embryo is phosphorylated to the mono-, di- and triphosphates. More than 80% is phosphorylated to guanosine triphosphate. The timing of increased guanosine phosphorylation correlates with a decrease in the acid-soluble GTP pools in the embryo, suggesting that increased guanosine kinase activity is a response to increased GTP demand. These results, in view of the importance of GTP in many cellular processes, imply a crucial role for guanosine kinase activation in GTP pool maintenance and cellular metabolism during early sea-urchin development. PMID- 3032631 TI - Colloidal gold labeling studies related to vascular and endothelial function, hemostasis and receptor-mediated processing of plasma macromolecules. PMID- 3032632 TI - Cyclic adenosine monophosphate stimulates biosynthesis and phosphorylation of the 26 kDa gap junction protein in cultured mouse hepatocytes. AB - Hepatocytes prepared from 18-day-old mouse embryos were grown in serum-free medium and reached confluence after two days in culture. The total amount of the 26 kDa gap junction protein decreased in these cells during the first 24 h in culture and increased again between day 1 and day 3 more than 10-fold. At day 3 a half-life time of 2.5 to 3 h was determined for the 26 kDa protein by [35S]methionine incorporation and immunoprecipitation using affinity-purified anti-26 kDa. Incorporation of [32P]orthophosphate into the 26 kDa protein of cultured hepatocytes was found at serine residues (98%) and tyrosine residues (about 2%). The addition of dibutyryl cyclic adenosine monophosphate (db cAMP) to the culture medium at day 2 had two effects: After 15 min the extent of phosphorylation of the 26 kDa protein increased 2.7-fold whereas the total amount of the 26 kDa protein increased only 1.2-fold. After 3 h of incubation with db cAMP, a 2.5-fold increase of the 26 kDa protein was noticed which was accompanied by a 3.2-fold increase in phosphorylation of serine residues. The effects of db cAMP on phosphorylation of the 26 kDa protein could be augmented or mimicked by the addition of isoproterenol, theophylline or forskolin to the culture medium of hepatocytes. In extracts of rat hepatocarcinoma MH1C1 cells and dog kidney MDCK cells, a phosphorylated 26 kDa protein can be immunoprecipitated using anti-liver 26 kDa. These results demonstrate that the gap junction 26 kDa protein can be posttranslationally modified by cAMP-dependent phosphorylation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3032633 TI - Detection of human papilloma virus infection in routine Papanicolaou-stained cervical smears. AB - Women at reproductive age were investigated cytologically for the presence of koilocytes in cervical scrapings to study the frequency of Human Papilloma Virus infections. Out of 2275 cervical smears, 51 (2.24%) revealed koilocytes. Koilocytes were more frequently associated with dysplasia and neoplasia than inflammation and negative smears. Higher frequency of HPV infection was observed in Hindu women as compared to Muslims. Blood stained or white discharge per vaginum is the commonest presenting symptom of the patients with HPV cervical lesions. The data observed in this study provide useful baseline information. PMID- 3032635 TI - Diagnostic problems of cervico-vaginal infections by HPV. AB - The diagnostic problems of cervico-vaginal infections by HPV have been considered. The colposcopic and cytological diagnoses have been compared with the histological one in order to evaluate the diagnostic reliability of the two methods of screening, and to seek evidence of those colposcopic and cytological aspects which, even if not pathognomic, are suggestive of HPV infection. The association between viral changes and CIN and the correlated problems have also been examined. The importance is underlined of screening and following up of HPV infections by the integrated use of colpocytology and colposcopy and of eventual aimed histologic checking as an improved prognostic evaluation, considering the potentiality of the evolutive risks of such infections. PMID- 3032634 TI - Human papillomavirus (HPV) infections involved in the neoplastic process of the uterine cervix as established by prospective follow-up of 513 women for two years. AB - A total of 513 women with cervical HPV infections have been followed-up since 1981 (mean 25.6 months) to establish the biological potential of HPV in cervical carcinogenesis. On each attendance, the patients were subjected to colposcopy accompanied by Papanicolaou (PAP) smears and/or punch biopsies. The latter were analysed for HPV particles on TEM, for the cytopathic changes of HPV, as well as for HPV structural proteins. The local immunocompetent cell (ICC) infiltrates are enumerated using ANAE-technique to define B-, MPS- and T cells, and monoclonal antibodies for T cell subsets, NK and K cells and Langerhans cells. HPV DNA typing was accomplished by Southern blot and spot hybridization using the DNA probes for HPV 6, 11, 16 and 18. Antibody titres for HSV were measured, and Chlamydia trachomatis isolations completed in cervical swabs. No correlation with the clinical course, e.g. regression (RE), persistence (PE), progression (PR) or recurrence (RC) of the HPV lesions could be established for the following factors; expression of HPV antigens, presence of HPV particles on TEM, Chlamydia in cervical swabs, the levels of HSV antibodies, and the levels of the ICCs. The OKT4+/OKT8+ cell ratio, however, was inversely correlated with PR, being most markedly reduced in recurrent lesions. Of the 513 lesions, 24.8% regressed, 59.8% remained persistent, and 14.1% progressed, 11.9% having been coned due to progression into CIS. So far, 1.1% of lesions have recurred after such a treatment. The progression rate was highest (45.5%) in HPV 16 lesions, followed by that (27.3%) in HPV 18 lesions, as contrasted to 0% and 13.3% for HPV 6 and 11, respectively. The results clearly confirm that cervical HPV infections are capable of progressing into CIS and thus show a natural history equivalent to that of classical CIN. The inherent potential of HPV 16 and HPV 18 lesions for clinical progression was clearly established, supporting the concept on HPV 16 and HPV 18 as the high risk HPV types in cervical carcinogenesis. PMID- 3032636 TI - Estrogen receptors and histopathological evaluation of human breast cancer. AB - The breast cancers with positive ER, the anaplastic low level and the node non involvement were identified with good prognostic values. The positive estrogen receptors in 52 primary breast carcinomas were correlated with either big or small tumors, with node involvement or without it and either with or without sinus histocitosis reaction. The vascular and lymph infiltration, the undifferentiated and half-differentiated carcinomas, the high stroma component and the high number of mitosis were other parameters with were opposed to positive ER tumors. The prevailing age of the patients was postmenopausal. PMID- 3032637 TI - Stress induced right ventricular dysfunction: an indication of reversible right ventricular ischaemia. AB - Stress induced changes in left ventricular ejection fraction are widely used in the detection and assessment of coronary artery disease. This study demonstrates that right ventricular dysfunction may also occur, and assesses its significance in terms of coronary artery anatomy. This study involved 14 normal subjects and 26 with coronary artery disease investigated by equilibrium radionuclide ventriculography, at rest and during maximal dynamic exercise. Mean normal resting right ventricular ejection fraction (RVEF) was 0.40 (SD 0.118), and all normal subjects increased RVEF with stress (mean delta RVEF + 0.13 SD 0.099). Mean delta RVEF in the subjects with coronary artery disease was significantly lower at 0.00 (SD 0.080), but there was overlap between the two groups. The largest falls in RVEF were seen if the right coronary artery was occluded without retrograde filling. In this subgroup with the most severely compromised right ventricular perfusion (nine subjects), RVEF always fell with stress, and mean delta RVEF was -0.08 (SD 0.050). There was no significant correlation between delta LVEF and delta RVEF, implying that the right ventricular dysfunction was due to right ventricular ischaemia, rather than secondary to left ventricular dysfunction. Stress induced right ventricular ischaemia can therefore be detected readily by radionuclide ventriculography. PMID- 3032638 TI - Uptake of technetium 99m MDP by hepatoblastoma. AB - Focal uptake of 99mTc-MDP was seen in a case of hepatoblastoma. The focal uptake corresponded to an area of calcification on CT, which was shown histologically to consist of osteoid with mineralization. The mechanism of uptake by the tumor in this case is likely to be the same as for skeletal uptake. PMID- 3032639 TI - Digoxin-like immunoreactive substance in serum of preterm and full-term neonates. AB - A significant serum level of digoxin-like immunoreactive substance (DLIS) (greater than or equal to 0.5 ng/ml) has been found in healthy full-term neonates, in prematurely born neonates as well as in full-term but small for gestational age neonates. Neither the babies nor their mothers had received digoxin therapy. On the first day of life, the incidence of serum levels of DLIS greater than or equal to 0.5 ng/ml in the three groups of neonates were respectively 64% (32/50), 42% (8/19) and 77% (10/13). Longitudinal measurements in preterm and small for gestational age neonates indicate a progressive disappearance of DLIS from their serum, none of them having a significant serum level at 21 days of age. As long as the chemical structure, origin and physiological properties of DLIS remain unknown, clinicians must be cautious in interpreting the serum levels of digoxin used for therapeutical purpose in neonates. PMID- 3032640 TI - Double-blind controlled study on the efficacy of sodium alginate (Gaviscon) in reducing gastroesophageal reflux assessed by 24 h continuous pH monitoring in infants and children. AB - We studied the effects of an alginate compound (Gaviscon) on the frequency and the duration of gastroesophageal reflux (GOR) episodes in children. Twenty infants and children with characteristic symptoms of GOR were divided at random into two groups which were given either Gaviscon (ten patients, mean age: 21 months) or a placebo (ten patients, mean age: 35 months). A continuous pH probe monitoring of the lower oesophageal third was performed in all the patients before and after 8 days of treatment. Before the trial, sensitive pH monitoring variables of acid reflux (Euler-Byrne index, percentage of total reflux time per 24 h, mean duration and percentage of reflux time during sleep, total number of reflux episodes per 24 h and number of reflux episodes per 2 h post-cibal periods) were abnormal in all the patients tested. The oesophagram revealed a GOR in 13 of the 20 patients; none of the children who underwent an endoscopy had evidence of oesophagitis. Episodes of regurgitation reported by the parents decreased during Gaviscon therapy while no clinical improvement was noticed in the placebo group. No adverse effects were observed. After 8 days of treatment with Gaviscon, results of all the pH monitoring variables were significantly (P less than 0.05) reduced between -35% and -61% of the initial values recorded. In the placebo treated group, the mean values remained little changes (-9.5 to +8.2% of initial values). These data suggest that Gaviscon may prove useful in the medical management of GOR in infants and children. PMID- 3032641 TI - The haematic thiamine level in the course of alcoholic neuropathy. AB - The specific roles of ethanol and malnutrition in the pathogenesis of neurological disorders of chronic alcoholism remain unclear. We measured plasmatic thiamine levels and erythrocyte transketolase activity in 30 alcoholics with peripheral neuropathy and in 4 with a Wernicke-Korsakoff (W-K) syndrome. Thiamine levels in the first group were comparable to those of normal subjects while a significantly lower concentration was found in W-K syndrome. Transketolase activity was lower for both groups in comparison with normal subjects. We suggest that a defect in thiamine utilization is involved in peripheral neuropathy of alcoholics, rather than a lack of thiamine itself. PMID- 3032642 TI - Progressive systemic sclerosis and nervous system involvement. A review of 14 cases. AB - Nervous system involvement in progressive systemic sclerosis (PSS) has been considered rare compared to other collagen diseases. We present 14 additional cases of PSS with neurological manifestations. Primary involvement of the peripheral nerves could be detected in 4 of 14 patients and is documented by electromyo- and electroneurographical examinations. Central nervous system (CNS) manifestations directly related to PSS are a rarity, which may reflect the lack of collagen in the brain, histological differences between cerebral and other arteries and the immunological particularity of the brain. There may have been a direct relationship between CNS involvement and PSS in only one patient presenting with an overlap-syndrome. PMID- 3032643 TI - Regional variation in high-density lipoprotein binding to human adipocyte plasma membranes of massively obese subjects. AB - Obesity is associated with significant changes in cholesterol and lipoprotein metabolism. High density lipoprotein (HDL) cholesterol is often reduced and adipose tissue cholesterol stores are increased in obese individuals. This prompted a study on the binding of HLD fractions (HDL2 and HDL3) to adipocyte plasma membranes obtained from massively obese subjects (BMI greater than 37 kg m 2) undergoing gastroplasty. Regional variation in HDL binding to these adipocyte plasma membranes was demonstrated. Membranes derived from the abdominal subcutaneous depot exhibited similar binding affinity (Kd) but higher binding capacity (Bmax) for HDL2 and HDL3 than that from the omental depot. There was significant inter-individual variation in Bmax but the amount of HDL2 or HDL3 bound to the two depots of the same individual was positively correlated (HDL2, r = 0.66, P less than 0.05; HDL3, r = 0.88, P less than 0.01). While HDL2 binding showed a higher affinity (lower Kd) than HDL3, a significant positive correlation existed between HDL2 and HDL3 binding to the same adipocyte membranes (r = 0.89, P less than 0.01). A significant inverse correlation (P less than 0.05) was also observed between HDL2 and HDL3 binding to adipocyte membranes and plasma HDL cholesterol concentration. These results suggest that adipose tissue is an important site of HDL metabolism and the subcutaneous fat depot may play a proportionally more significant role due to its higher HDL binding capacity. It is further suggested that increased HDL binding and metabolism by the expanded adipose tissue mass may contribute to reduced plasma HDL-cholesterol levels frequently associated with obesity. PMID- 3032644 TI - Production of 5-lipoxygenase pathway metabolites by peripheral leucocytes in capillary leak syndrome (Clarkson disease). AB - Periodic systemic capillary leak syndrome (Clarkson disease) is characterized by unexplained attacks of a marked increase in capillary permeability. As leukotrienes, derived from arachidonic acid via the 5-lipoxygenase pathway, enhance capillary permeability, we studied arachidonate metabolism in leucocytes of a patient with capillary leak syndrome. Leucocyte-platelet suspensions, prepared from blood collected from the patient during asymptomatic periods (n = 11) produced greater amounts of 5-hydroxyeicosatetraenoic acid (5-HETE) than control suspensions (P less than 0.05). Peripheral leucocytes, collected during attacks (n = 3) and studied without addition of A23187 released LTB4 in vitro but not sulphidopeptides leukotrienes. This result was never observed with leucocytes from control subjects or from the patient out of a crisis. These results suggest that in the patient, peripheral leucocytes could be stimulated by an unknown, as yet to be determined, endogenous factor to produce more 5-HETE and LTB4. Whether LTB4 plays a pathogenic role in the capillary leakage remains to be determined. PMID- 3032645 TI - Role of alpha-adrenoceptors for adipocyte size in man. AB - Catecholamines have dual effects on lipid mobilization in man. Lipolysis is stimulated via beta-adrenoceptors and inhibited via alpha 2-adrenoceptors. The relationship between fat-cell size and catecholamine-induced lipolysis (expressed per cell surface area) was investigated in vitro in subcutaneous adipocytes of thirty-five non-obese subjects between 1 month and 45 years of age. Fat-cell volume showed a positive correlation with noradrenaline-induced lipolysis (r = 0.7). Furthermore, fat-cell size showed a negative correlation with the alpha 2 effect of noradrenaline (r = 0.8) but no correlation with the beta-effect of the hormone (r less than 0.1). Although age showed a positive correlation with noradrenaline-induced rate of lipolysis (r = 0.6) it did not contribute to the relationship between cell size and the catecholamine effect. There was no relationship between adipocyte size and the basal (unstimulated) rate of lipolysis (r less than 0.1). In conclusion, ageing in non-obese subjects is associated with enlargement of fat-cell size and enhancement of the lipolytic effect of catecholamines. The latter is due to diminished alpha 2-anti-lipolytic effect of the hormones. This modulation of the alpha 2-adrenoceptor activity may be of importance for the regulation of adipocyte size in man. PMID- 3032646 TI - Haematological and hormonal responses to dynamic exercise in patients with chronic airway obstruction. AB - The influence of exercise on hormonal and total white blood cells (WBC), lymphocytes (L). Granulocytes (GR), and platelet (P) count responses was studied in: twenty-five patients with chronic airway obstruction (CAO, 47 +/- 1.8 years, mean +/- SEM) and thirteen normal subjects (N, 36 +/- 2.6 years). They performed a submaximal (40 W) and a maximal exercise (VO2max). Arterial blood samples were taken at rest, 40 W, and VO2max. [H+], PaCO2, PaO2 haematocrit (Hct), [Hb], P, total platelet volume (TPV), WBC, GR, L, and total red blood cells (RBC) were measured. At rest, WBC, GR, P and TPV were higher in CAO patients, whilst PaO2 and cortisol were lower. At 40 W, when compared to values obtained at rest, WBC, GR, L, P and TPV were increased in both groups; WBC, GR, P and TPV were higher in CAO patients. VO2max of CAO patients represented 54% of that of controls. At VO2max, Hct, [Hb] and RBC were approximately 10% higher than at rest in both groups, whilst changes were more significant in normals for WBC (CAO = 55%, N = 76%), lymphocytes (CAO = 83%, N = 105%), GR, (CAO = 37%; N = 51%), platelets (CAO = 23%, N = 29%), TPV (CAO = 25.4%, N = 35%), [H+] (CAO = 43%, N = 38%) and ACTH (CAO = 82%, N = 139%). PaO2 and cortisol did not differ between groups. PaCO2 and platelets however, were higher in the CAO group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3032647 TI - Hybridomas producing human monoclonal antibodies against varicella-zoster virus. AB - Hybridomas producing human monoclonal antibodies (mAb) against varicella-zoster virus (VZV) were generated by fusing human splenic lymphocytes with mouse myeloma cells. Before cell fusion, lymphocytes were stimulated in vitro with viral antigens and pokeweed mitogen. This combination synergistically increased the generation of VZV-specific hybridomas. Five established hybridomas have been stably producing mAb for at least 9 months. These mAb, designated V1, V2, V6, V8 and V9, were of the IgG1, lambda isotype. They bound to all 6 tested VZV strains but not to other herpes viruses, with the exception that V1 bound to herpes simplex virus (HSV) as well as VZV. Immunoprecipitation analysis showed that V1, V6 and V9 recognized glycoprotein gpII, whereas V2 and V8 recognized gpI. In addition, V1 reacted with the gB glycoprotein of HSV. All these mAb neutralized viral infectivity. The neutralizations by V2 and V8 were more effective and more complement dependent than those by V1, V6 and V9. Immunofluorescence tests revealed that all these mAb bound to the surface membrane of VZV-infected cells. These results suggest that cell fusion between in vitro stimulated lymphocytes and mouse myeloma cells is a reliable method for the generation of hybridomas capable of stable production of human mAb. The human mAb thus developed may provide a new means of passive immunization of humans against VZV infection. PMID- 3032648 TI - Interactions between hepatitis B virus and polymeric human albumin. I. Production of monoclonal anti-idiotypes (anti-anti-polymeric human albumin) which recognize hepatitis B virus surface antigen. AB - In an attempt to characterize the polymeric human albumin (polyHSA) receptor expressed on hepatitis B virus and hepatocytes, we have used a human anti-polyHSA IgG to generate monoclonal anti-idiotypes (anti-Id) which bear the internal image of polyHSA and mimic its binding activity. Two monoclonal anti-Id antibodies, 63.14 and 70.F9, were strongly reactive in both radioimmunoassay and enzyme linked immunosorbent assay (ELISA) with the F(ab')2 of the immunogen as well as with purified hepatitis B surface antigen (HBsAg) expressing various subtypes. The specificity of the binding of anti-Id to HBsAg was confirmed in direct ELISA and by Western blot analysis. These experiments also showed that the anti-Id bind to a site expressed on the major 24-kDa protein of HBsAg particles, and that this recognition is specifically inhibited by polyHSA. Experiments on cellular staining and radioimmunoprecipitation on HBsAg-positive and -negative cell lines showed that the anti-Id recognize intracellular HBsAg but not other liver cell proteins, including the putative polyHSA receptor. These data indicate, therefore, that the monoclonal anti-Id mimic the binding activity of polyHSA and recognize its binding site on the virus. The inability of both anti-Id to react with the hepatocyte surface suggests either the absence of a specific hepatic polyHSA receptor or the expression of one with a different configuration. PMID- 3032649 TI - Effects of pertussis toxin on the alpha 2-adrenoceptor-inhibitory GTP-binding protein-adenylate cyclase system in rat brain: pharmacological and neurochemical studies. AB - Behavioral excitement and the increase in locomotion were observed in male adult rats four days after an intraventricular injection of 5 micrograms pertussis toxin (IAP). Clonidine (100 micrograms/kg s.c.)-induced locomotor hypoactivity was not observed in animals pretreated with 1 and 5 micrograms IAP. IAP caused a significant (P less than 0.05) decrease in the KD value of [3H]clonidine binding and enhanced GTP (1 microM)-induced decrease in the binding to cortical membranes from rat brain. In addition, the inhibition of adenylate cyclase induced by alpha 2-receptor stimulation (100 microM adrenaline plus 100 microM propranolol) was completely suppressed in the cerebral cortical membranes by IAP pretreatment. It is suggested that the system consisting of alpha 2-receptor, the inhibitory GTP binding protein (Ni) and adenylate cyclase inhibits some animal behaviors and cyclic AMP formation. Moreover, IAP seems to inactivate Ni, subsequently producing behavioral excitement and it inhibits clonidine-induced sedation. PMID- 3032651 TI - (Na+,K+)-ATPase and noradrenergic function: effects of chronic ethanol. AB - The experiments in this paper examined interactions between ethanol and repeated noradrenergic stimulation in vivo on regulation of (Na+,K+)-ATPase. The increase in ouabain binding and K+-phosphatase activity associated with (Na+,K+)-ATPase in rats treated with repeated yohimbine injections was prevented by chronic ethanol. Ethanol did not affect the yohimbine-induced alterations in noradrenergic receptor binding or in content of the norepinephrine metabolite 3-methoxy-4 hydroxyphenylglycol, showing that prevention of noradrenergic stimulation of (Na+,K+)-ATPase was not caused by a decrease in availability of norepinephrine. In addition, norepinephrine depletion with the neurotoxin DSP4 did not prevent the increases in (Na+,K+)-ATPase indices during chronic ethanol treatment, showing that they did not result from increased norepinephrine exposure. These results suggest that chronic ethanol reduces sensitivity of (Na+,K+)-ATPase to norepinephrine in vivo, possibly as a consequence of membrane effects of ethanol tolerance. PMID- 3032650 TI - A comparison of the neurochemical and behavioral effects of clenbuterol and desipramine. AB - Because both long-term adrenoceptor agonist administration and antidepressant treatment in animals down-regulate CNS beta-adrenoceptors and attenuate brain adenylate cyclase activity, beta-adrenoceptor agonists may also possess antidepressant properties. We compared the effects of the centrally acting beta adrenoceptor agonist clenbuterol (5, 10 and 35 mg/kg per day), and the combination of propranolol (5 mg/kg per day) and clenbuterol (10 mg/kg per day), with desipramine (15 mg/kg per day) on forced swim test performance and on cortical beta-adrenoceptors in rats following 7 days of drug administration. Desipramine (15 mg/kg per day), and clenbuterol (10 and 35 mg/kg per day, but not 5 mg/kg per day) both significantly reduced immobility in the forced swim test. Frontal cortex beta-adrenoceptors were significantly down-regulated after desipramine and all 3 doses of clenbuterol. The co-administration of propranolol (5 mg/kg per day) blocked both the reduction in immobility and down-regulation of cortical beta-receptors induced by clenbuterol (10 mg/kg per day). Propranolol (5 mg/kg per day) alone up-regulated frontal cortex beta-adrenoceptors, but had no significant effect on swimming performance. These data suggest that the physiological consequences of beta-adrenoceptor down-regulation are important in the mechanism of action of antidepressants. The results also suggest that clenbuterol may be useful in the treatment of depression. PMID- 3032652 TI - Effect of dB-c-AMP and forskolin on the 45Ca influx, net Ca uptake and tension in rabbit aortic smooth muscle. AB - The effects of dibutyril-adenosine-3',5'-cyclic monophosphate (dB-c-AMP) and forskolin on aortic tension and 45Ca influx were measured. dB-c-AMP reduced both the rate of force development and the maximal tension achieved in solutions containing various K+ concentrations. Stimulated 45Ca influx was also reduced, however, to a lesser extent than was the tension. Forskolin showed more marked effects of a similar nature. Thus, both these agents which increase intracellular c-AMP caused a rightward shift in the curve expressing force (ordinate) as a function of Ca2+ influx (abscissa). Consequently, we found that dB-c-AMP stimulated more net Ca to be taken up by sarcoplasmic reticulum (SR) at the same influx rate. The conclusion that c-AMP produced these effects by stimulating Ca uptake into the superficial SR was supported by the finding that dB-c-AMP increased the amount of Ca taken up into a caffeine releasable fraction. PMID- 3032653 TI - d(CH2)5Tyr(Me)-[Orn8]vasotocin, a potent oxytocin antagonist, antagonizes penile erection and yawning induced by oxytocin and apomorphine, but not by ACTH-(1-24). AB - Intraventricular (i.c.v.) injection of d(CH2)5-Tyr(Me)-[Orn8]vasotocin, a potent oxytocin antagonist, antagonized in a dose-dependent manner (10-100 ng) penile erection and yawning induced by the systemic injection of apomorphine (80 micrograms/kg s.c.) or by the i.c.v. injection of oxytocin (30 ng). In contrast, the oxytocin antagonist, even at the dose of 10 micrograms, did not modify penile erection and yawning induced by the i.c.v. injection of ACTH-(1-24). These results suggest that apomorphine, but not ACTH-(1-24), induce penile erection and yawning by releasing oxytocin in some brain area. PMID- 3032654 TI - Autoradiographic localisation of NPY receptors in rabbit kidney: comparison with rat, guinea-pig and human. AB - In vitro labelling of neuropeptide Y binding sites with 125I-NPY was performed in rabbit kidney sections. Autoradiographic and histochemical techniques localised these binding sites to the proximal convoluted tubules and the vascular smooth muscle. In comparison, 125I-NPY binding was absent from the rat, guinea pig and human kidney. Possible physiological roles for the putative NPY receptors in rabbit renal function are discussed. PMID- 3032655 TI - The dopamine hypothesis of opiate reward challenged. AB - The dopamine hypothesis of opiate reward was directly tested by blocking dopamine receptor systems with haloperidol. Haloperidol was administered either systemically or locally into different brain areas with dopaminergic terminals in rats allowed to initiate and maintain intravenous heroin self-administration. Haloperidol treatment did not block heroin reward. It is concluded that dopamine is not critically involved in opiate reward and that multiple endogenous reward systems are present in the brain. PMID- 3032656 TI - Effects of agents which interact with central benzodiazepine binding sites on stress-induced ultrasounds in rat pups. AB - Compounds of different chemical classes, with affinity for central benzodiazepine receptors, were investigated for effects on handling-induced ultrasonic cries in rat pups. Diazepam and clobazam inhibited ultrasounds and impaired motor performance at higher doses. Suriclone showed a similar profile to diazepam but premazepam clearly separated ultrasound inhibition from motor impairment. ZK 91296, CGS 9896, RU 39419 and RU 43028 inhibited ultrasounds with a lower maximal response but induced little or no motor incoordination. CL 218872 inconsistently inhibited sounds with no motor impairment and PK 8165 and PK 9084 had no consistent effects. Benzodiazepine antagonists weakly inhibited (RU 40410) or did not affect (Ro 15-1788 or CGS 8216) ultrasounds alone but fully (Ro 15-1788 or CGS 8216) or partially (RU 40410) antagonised the effects of benzodiazepines. Inverse agonists FG 7142, methyl-beta-carboline-3-carboxylate (BCM) and DMCM tended to increase ultrasounds alone, particularly when less stressful handling stimuli were used, and antagonised benzodiazepines. This procedure detects the differing behavioural effects of benzodiazepines receptor ligands and is proposed as a simple model of anxiety. PMID- 3032657 TI - A single dose of FG 7142 causes long-term increases in mouse cortical beta adrenoceptors. AB - There is evidence that central monoamines are involved in the actions of benzodiazepines. We have investigated the effects of a single dose of the benzodiazepine partial inverse agonist, FG 7142, on radioligand binding to alpha 2- and beta-adrenoceptors in mouse cerebral cortex. We found that seven days after a single injection of FG 7142 there was a large increase in the density of beta-adrenoceptors. This rise was not detectable either 15-30 min, or 24 h after the injection and no statistically significant changes in alpha 2-adrenoceptor binding were found at any of these times. Administration of the benzodiazepine antagonist Ro 15-1788 at the same time as FG 7142 prevented the rise in beta adrenoceptor density. We discuss the possibility that the beta-adrenoceptor upregulation is related to the behavioural effects of FG 7142. PMID- 3032658 TI - Repeated electroconvulsive shock prevents increased neocortical beta 1 adrenoceptor binding after DSP-4 treatment in rats. AB - Repeated electroconvulsive shock (ECS) was administered to rats previously injected with DSP-4 (N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine hydrochloride), a noradrenergic neurotoxin. The normal increase in neocortical beta 1 adrenoceptor binding caused by noradrenaline depletion was effectively prevented by ECS. This finding suggests that the plasticity of the beta 1-adrenoceptor may be partially independent of endogenous noradrenaline concentration. Additionally, functional noradrenergic neurons are not necessarily a critical requirement for the antidepressant effect of electroconvulsive treatment in humans. PMID- 3032659 TI - Comparison of GABA-induced responses in various segments of the guinea-pig intestine. AB - In preloaded, isolated segments of duodenum, jejunum, ileum and colon, ethylenediamine (EDA) caused a 3-mercaptopropionic acid (3-MPA)-sensitive release of [3H]GABA. Both EDA and GABA caused a contraction and a delayed 'after relaxation' in the ileum, but only a delta-aminovaleric acid (DAVA)-sensitive relaxation in other intestinal segments; these actions of EDA were reduced by 3 MPA and therefore due to release of endogenous GABA. Baclofen induced a DAVA sensitive relaxation in all tissues. Evidently, GABA modulates cholinergic excitation of the smooth muscle at all levels of the intestine. PMID- 3032660 TI - Pertussis toxin does not inhibit the alpha 1-adrenoceptor-mediated effect on inositol phosphate production in the heart. PMID- 3032661 TI - Protein kinase C inhibits TRH-stimulated phosphoinositide hydrolysis in GH3 cells. AB - The GH3 pituitary tumor line expresses TRH receptors that stimulate phosphoinositide hydrolysis and hormone secretion. After protein kinase C was identified in GH3 cells by direct labeling with [3H]phorbol dibutyrate (PDB), the response to phorbol ester and TRH pretreatment on subsequent TRH-stimulated inositol phosphate (IP) accumulation was found to be inhibitory. Both phorbol myristate acetate (PMA) and PDB were effective in this regard at low nM concentrations within a few minutes, whereas phorbols that do not stimulate protein kinase C were without effect. Furthermore, the mono-, bis- and tris phosphate forms of IP were all reduced by an average of 30-40% after 5 min of PMA. TRH concentration-response studies indicated a clear change in TRH efficacy induced by PMA. Finally, preincubation with TRH itself was also capable of reducing the subsequent response to TRH. Because TRH receptor action is thought to activate protein kinase C by producing diacylglycerol, these data indicate a negative feedback system via protein kinase C operative during continuous exposure to TRH in GH3 cells. PMID- 3032662 TI - Photoaffinity labeling of a novel benzodiazepine binding protein in rat brain. AB - Photoaffinity labeling experiments were conducted with [3H]clonazepam to investigate the possible existence of a benzodiazepine binding site that is distinct from the central- and peripheral-type benzodiazepine binding sites. These studies demonstrate the presence of a novel benzodiazepine binding protein in rat brain with a relative mobility of 65,000 daltons that binds benzodiazepines stereoselectivity in the high nanomolar-low micromolar concentration range. PMID- 3032663 TI - Cellular regulation of poly ADP-ribosylation of proteins. II. Augmentation of poly(ADP-ribose) polymerase in SV40 3T3 cells following methotrexate-induced G1/S inhibition of cell cycle progression. AB - SV40-3T3 cells were exposed in monolayer cultures to 5 X 10(-7) M methotrexate (MTX), that inhibited thymidylate synthetase, arrested cell growth without cell killing in 24 h and did not induce single- (ss) or double-strand (ds) breaks in DNA. Following 24, up to 72 h, the poly(ADP-ribose) polymerase content of attached cells was induced by 5 X 10(-7) M MTX and the augmentation of the enzyme increased with the time of exposure to the drug. Inhibition of protein or RNA synthesis abolished augmentation of enzymatic activity; so too did the initiation of maximal cell growth by thymidine + hypoxanthine, by-passing the inhibitory site of MTX. Isolation of the ADP-ribosylated enzyme protein by gel electrophoresis identified poly(ADP-ribose) polymerase protein as the molecule that was induced by 5 X 10(-7) M MTX. Under identical conditions, the poly(ADP ribose) polymerase induction in 3T3 cells could not be demonstrated. A possible cell-cycle-dependent biosynthesis of the enzyme protein is proposed in SV40 3T3 cells. PMID- 3032664 TI - Actions of MPTP and MPP+ on synaptic transmission in guinea-pig hippocampal slices. AB - MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) causes a Parkinson's disease like syndrome in man, monkeys, and mice. We studied the effects of MPTP and its metabolite, MPP+, on neuronal properties and synaptic transmission in isolated slices of guinea-pig hippocampus using intra- and extracellular recording methods. Addition of MPTP to the superfusate (50 to 100 microM) produced the following effects: Excitatory postsynaptic potentials and extracellularly recorded population spikes, evoked by stimulation of the Schaffer collaterals were increased in amplitude during the application period (30 min). Within 30 min of washing in normal solution, synaptic transmission was blocked, although axonal population action potentials could still be elicited. The block of synaptic transmission was prevented by prior incubation in pargyline, an inhibitor of monoamine oxidase. The membrane potential and resistance of single pyramidal neurons were virtually unaffected; action potentials elicited by depolarizing intracellular current pulses were also unchanged. MPP+ (50 microM) blocked synaptic transmission during the application period by a pargyline-in-sensitive mechanism. These results suggest that MPP+ blocks synaptic transmission in the hippocampus at a presynaptic site. This effect may be relevant for the acute action of MPTP and may provide some insight into its chronic action on nigrostriatal neurons. PMID- 3032665 TI - Developmental expression of the muscle-specific isozyme of phosphoglycerate mutase in human muscle cultured in monolayer and innervated by fetal rat spinal cord. AB - The electrophoretic pattern of phosphoglycerate mutase of adult innervated normal human muscle is composed predominantly of the muscle-specific isozyme, whereas the electrophoretic pattern of aneurally cultured human muscle is composed only of the brain-specific isozyme. We studied the transition of the isozymes (phosphogluterate mutase) in human muscle cultured in monolayer and innervated for 20 to 83 days by rat embryo spinal cord explants. In this culture system, regions of innervated muscle fibers in close proximity to the ventral part of the spinal cord explant continuously contracted and the contractions were reversibly blocked by 1 mM d-tubocurarine. In those innervated cultured human muscle fibers, the total activity of phosphoglycerate mutase was increased and the muscle specific isozyme was expressed. The amount of muscle-specific isozyme directly correlated with the duration of innervation. This study demonstrated that expression of the gene for the muscle-specific isozyme of phosphoglycerate mutase in human muscle cultured in monolayer is influenced by de novo innervation. PMID- 3032666 TI - Babesia bovis: molecular and biological characteristics of cloned parasite lines. AB - An in vivo limiting dilution technique was used to produce several Babesia bovis cloned lines with which to study the basis of virulence and immunogenicity in this parasite. DNA hybridization using a cloned DNA fragment from the BabR locus demonstrated that the cloned lines were a more restricted genetic population than the parent strain. Biosynthetic labeling and immunoprecipitation studies indicated that the cloned lines differed from each other and from the parentals in the expression of a small number of polypeptides and antigens. Animal trials with three of the lines demonstrated that the parental line contains both virulent and avirulent parasite populations, at least three of which are not tick transmissible, and that while the lines do provide significant protection against heterologous challenge, they may not give as effective protection as the parental line. These experiments demonstrated the existence of subpopulations with distinctive molecular and biological properties, providing evidence that the attenuation process is based on the selection of preexisting parasite subpopulations combined with the ability of these parasites to vary genetically. PMID- 3032667 TI - Long-term adrenergic beta-action decreases and alpha-action enhances corticosterone levels in rats. AB - A 20 h hyperadrenalinemia in rats was produced by subcutaneously implanted A retard tablets with an output rate of 1.8 micrograms/min/250 g. This caused a moderate (6 h, 20 h) to expressed (12 h) rise in Corticosterone. Concomitant beta blockade leads to equal (12 h, 20 h) or even more expressed (6 h) enhancement of plasma corticosterone, while A + alpha-blockade lowers those levels significantly against A or A + Prop treated animals. At 6 and 20 h they are even significantly lower than control values. We therefore conclude that enhanced alpha-adrenergic influence increases and beta-adrenergic influence decreases plasma Corticosterone levels in rats. PMID- 3032668 TI - Stabilisation of collagen by betel nut polyphenols as a mechanism in oral submucous fibrosis. AB - Treatment of reconstituted collagen fibrils and pieces of rat dermis with the crude extract, purified tannins or (+)-catechin from betel nut (Areca catechu) increases their resistance to both human and bacterial collagenases in a concentration-dependent manner. These tanning agents may stabilise collagen in vivo following damage to the oral epithelium, and promote the sub-epithelial fibrosis which occurs in betel nut chewers. PMID- 3032669 TI - A major metabolite of delta 1-tetrahydrocannabinol reduces its cataleptic effect in mice. AB - The results described here demonstrate that THC-induced catalepsy in mice can be substantially inhibited by the prior administration of delta 1-THC-7-oic acid, the major metabolite of THC in most species including humans. This raises the possibility that the intensity and duration of action of THC may depend to a large degree on the levels of this metabolite at the sites of action. PMID- 3032671 TI - Evidence for the presence of virus in clonal insulin-producing RINm5F cells. AB - The ultrastructural morphology of the clonal insulin-producing cell line, RINm5F, was investigated. Virus-like particles, probably C type viruses, were identified both intra- and extracellularly. Because these particles could not be found in the original transplantable tumor, it is probable that viruses were induced at some later stage in the development of the RINm5F cell line. All investigators using the RINm5F cells should be aware of the fact that these cells may contain one or several types of viruses, and of the possibility that these particles may interfere with a variety of cellular functions. PMID- 3032670 TI - Effect of acute administration of delta 1-tetrahydrocannabinol on beta-endorphin levels in plasma and brain tissue of the rat. AB - Acute treatment with delta 1-tetrahydrocannabinol (delta 1-THC) elevated the concentration of beta-endorphin-like immunoreactivity (beta-ELIR) in plasma and in the hypothalamus, but not in the hippocampus of rats habituated to the injection procedure. These effects were not obtained with the psychotropically inert analog of delta 1-THC, cannabidiol. In animals that had not been habituated to the injection procedure, placebo treatment induced a decrease in hippocampal beta-ELIR. PMID- 3032672 TI - Encephalomyocarditis (EMC) virus-induced diabetes mellitus prevented by Corynebacterium parvum in mice. AB - Corynebacterium parvum prevented the development of encephalomyocarditis virus induced diabetes in mice, when it was given 3-14 days before the virus infection. This treatment inhibited virus replication in the pancreas of the infected mice at an early stage of the infection. PMID- 3032673 TI - GMP-stimulation of the cyanide-insensitive mitochondrial respiration in heat shocked conidia of Neurospora crassa. AB - In mitochondria of heat-shocked conidia of Neurospora exogenous NADH and succinate were oxidized mainly via the alternative, hydroxamate-sensitive pathway (70%) and only 30% via the cytochromic, cyanide-sensitive pathway which was predominant in untreated conidia; the alternative oxidase pathway was markedly stimulated by guanosine 5'-monophosphate (GMP). PMID- 3032674 TI - Opioid peptides. Biological study of [D-MetO2] dermorphin analogues in relation to the plurality of opioid receptors. XI. AB - The opioid activity pattern of 8 dermorphin tetrapeptides, W-Tyr-D-MetO-Phe-Xaa-Y (W = H, H2N-C = (NH); Xaa = Gly, 2-aminoethanol, sarcosine; Y = NH2, NH-alkyl), was determined in guinea-pig ileum (GPI) preparations and in brain binding assays and compared with their antinociceptive potency in mice. Almost all modifications increased potency on the GPI test as well as the antinociceptive action of the parent H-Tyr-D-Ala-Phe-Gly-NH2. Dermorphin, H-Tyr-D-Ala-Phe-Gly-Tyr-Pro-Ser-NH2, and related tetrapeptide analogues show negligible K-binding activity and, like morphine, possess a higher affinity to mu- than delta-receptors. Nevertheless the correlation of analgesia with the effects on GPI and binding data separate the peptides from morphine. For example, in comparison with the opiate alkaloid H-Tyr D-MetO-Phe-Gly-ol and H-Tyr-D-MetO-Phe-Gly-NH2 displayed a lower affinity for mu sites, a moderately higher potency on GPI but an exceptionally stronger analgesia, being respectively 350 and 1500 times as potent an analgesic as morphine. This may involve different subpopulations of opioid mu-receptors. PMID- 3032675 TI - Proton magnetic resonance of the bovine spleen green heme-protein. AB - The ferric spleen green heme-protein exhibits hyperfine-shifted proton resonances between 90 and 20 ppm for the high-spin resting form and the chloride complex, and between 46 and -9.4 ppm for the low-spin nitrite complex. The proton NMR spectral profile of the enzyme is similar to that of lactoperoxidase, but different from those of common heme-proteins. The appearance of a resonance at 76 ppm in the ferrous enzyme shows the presence of a proximal histidine residue linked to the iron. The proton relaxation rates of bulk water indicate that chloride binds to the sixth position of the iron in the chloride complex of the enzyme. PMID- 3032676 TI - Evidence for N coordination to Fe in the [2Fe-2S] center in yeast mitochondrial complex III. Comparison with similar findings for analogous bacterial [2Fe-2S] proteins. AB - Yeast mitochondrial complex III contains a subunit with a [2Fe-2S] cluster (the Rieske center) that has unusual physical and chemical properties. For apparently similar centers isolated from bacteria, it has been shown by electron nuclear double resonance (ENDOR) and electron spin echo envelope modulation (ESEEM) measurements that these [2Fe-2S] centers are coordinated by at least one and probably two nitrogen ligands. This work describes similar ENDOR and ESEEM studies on the intact mitochondrial complex. We find that this [2Fe-2S] cluster exhibits ESEEM and ENDOR properties that appear to be indistinguishable from those observed with the isolated bacterial systems. Furthermore, changes in EPR lineshape that occur as complex III is progressively reduced are not accompanied by any changes in the nitrogen coupling parameters. This spectroscopic evidence for nitrogen coordination is supported by published sequence data on four Rieske iron-sulfur subunits. It seems likely that this is a general characteristic of such [2Fe-2S] redox active centers. PMID- 3032677 TI - De novo CDP-choline-dependent glycerophosphorylcholine synthesis and its involvement as an intermediate in phosphatidylcholine synthesis. AB - The activity of CDP-choline-dependent glycerophosphorylcholine synthetase (CDP choline:sn-3-glycerophosphate cholinetransferase), a newly discovered enzyme involved in the recently proposed pathways of acyl-specific phosphatidylcholine synthesis, is reported in rat liver. Endogenous CDP-choline, synthesized via the CMP-driven back reaction of phosphorylcholine transferase, is also shown to be a choline donor for this glycerophosphorylcholine synthetase. The function of glycerophosphorylcholine as an intermediate in phosphatidylcholine synthesis is demonstrated by specific isotope trapping whereby unlabelled glycerophosphorylcholine inhibited label incorporation from sn-[14C]glycerol-3 phosphate into phosphatidylcholine in mouse gastrocnemius, a tissue that is essentially devoid of the cytidine pathway for phosphatidylcholine synthesis and uses a non-allelic glycerophosphorylcholine synthetase (exogenous PC:sn-3 glycerophosphate cholinetransferase) in the synthesis of glycerophosphorylcholine. PMID- 3032678 TI - Adenovirus infection reverses the antiviral state induced by human interferon. AB - HeLa cells treated with human lymphoblastoid interferon do not synthesize poliovirus proteins. The antiviral state against poliovirus is reversed if cells are previously infected with adenovirus type 5. A late gene product seems to be involved in this reversion, since no effect is observed at early stages of infection or in the presence of aphidicolin. PMID- 3032679 TI - A commentary on the inhibition by retinoids of leukotriene B4 production in leukocytes. AB - The order of potency of retinoids as inhibitors of A23187-induced production of leukotriene B4 (LTB4) in human polymorphonuclear leukocytes (PMN) was retinoic acid greater than retinal greater than retinol. However, the conversion of exogenous arachidonate (AA) to LTB4 by PMN homogenates was inhibited in the rank order retinol greater than retinal much greater than retinoic acid. The agreement between active concentrations of retinol in these two systems is consistent with this compound acting directly to inhibit AA metabolism: this is not so for the other retinoids. The order of potency for inhibition of phorbol dibutyrate (PDBu) stimulated superoxide (O-2) production in HL60 granulocytes was retinol greater than retinoic acid much greater than retinal (inactive); neither retinol nor retinal displaced [3H]PDBu from HL60 cells. We conclude that inhibition of LTB4 production by retinoic acid and retinal is neither through inhibition of AA metabolism nor through inhibition of protein kinase C. PMID- 3032680 TI - Decrease of cellular ATP by dihexanoylglycerol may limit responses to protein kinase C activation. AB - 1,2-sn-Dihexanoylglycerol (HHG) reduced the ATP content of HL-60 cells. This concentration-related (10-100 microM) effect reached a maximum of over 90%, was enantiomerically specific and not accompanied by release of lactate dehydrogenase. Oleoylacetylglycerols (3-100 microM) had no effect on ATP levels while phorbol dibutyrate (PDBu, 0.01-1 microM) decreased ATP content of HL-60 cells by up to 40%. Responses stimulated by HHG became limited as the concentration was increased above 10 microM, this being manifest as either a low maximum response compared to PDBu (superoxide release) or a bell-shaped concentration-effect curve (degranulation). HHG (30-100 microM) inhibited PDBu stimulated superoxide release, this inhibition being enantiomerically specific. It is probable that the effect of HHG on ATP content impairs cellular responses mediated through protein kinase C activation. PMID- 3032682 TI - Expression of a truncated v-myb product in transformed chicken embryo fibroblasts. AB - Transformed cells have been isolated after transfection of chicken embryo fibroblasts (CEF) with the DNA of a recombinant clone (KXA 3457) in which the v myb sequences are flanked by the two AMV-LTRs. Abnormal myb-specific RNA species and myb-related polypeptides were found to be expressed in these cells, suggesting that transformation of CEF by v-myb might require alterations of the oncogene product. PMID- 3032681 TI - Homology between bacterial DNA and bovine mitochondrial DNA encoding cytochrome c oxidase subunit III. AB - A segment of mitochondrial DNA encoding the bovine cytochrome c oxidase subunit III gene was isolated and inserted into an Escherichia coli plasmid vector. A 556 base pair fragment of the insert DNA representing about 70% of the 3'-end of the subunit III gene was used to search for homology with bacterial DNA from strains that contain heme aa3-type cytochrome c oxidases. Bacillus subtilis, Thermus thermophilus, and PS3 DNAs all showed strong hybridization to the probe, whereas Paracoccus denitrificans and Rhodopseudomonas sphaeroides DNAs showed only weak hybridization to the probe, even under low stringency conditions. PMID- 3032683 TI - A novel superoxide radical generator in heart mitochondria. AB - Experimental evidence is presented demonstrating the existence of a potent O2.- source in heart mitochondria. The novel O2.- generator is more active than any other known mitochondrial O2.- generator and also exhibits a higher affinity for molecular oxygen. In contrast to mitochondrial O2.- sources reported previously [(1974) FEBS Lett. 42, 68-72; (1978) Eur. J. Biochem. 82, 563-567], the O2.- generator described in this paper is not involved in energy-linked respiration. Superoxide radicals from this source require NADH to initiate their generation, and the radicals formed are released entirely into the extramitochondrial space. NADH-related O2.- generation was also observed with the solubilized exogenous NADH oxidoreductase of heart mitochondria, an enzyme recently described [(1987) Eur. J. Biochem., submitted]. This finding together with the lack of an NADH dependent O2.- source in liver mitochondria suggests that the novel O2.- generator and the exogenous NADH oxidoreductase of heart mitochondria are identical. PMID- 3032684 TI - ESR signals from stimulated and resting porcine blood neutrophils. AB - The NADPH oxidase in neutrophils was specifically solubilized from membrane vesicles of porcine blood neutrophils and rapidly concentrated by immunoprecipitation with cross-reacting anti-P-450 reductase IgG. The precipitates from both myristic acid-stimulated and resting cells contained one third of the cytochrome b-558 and were slightly contaminated with myeloperoxidase. The immunoprecipitate from stimulated cells gave rhombic high spin ESR signals of a heme at g = 6.47 and 5.49, which were insensitive to KCN, whereas the preparation from resting cells did not give these signals. The rhombic high-spin signals are discussed in view of the participation of cytochrome b-558 in the NADPH oxidase system. PMID- 3032685 TI - BliI, a restriction endonuclease from Bacillus licheniformis. AB - From Bacillus licheniformis a site-specific restriction endonuclease, named BliI, has been purified and characterized. BliI was able to digest lambda DNA at pH 9.1 over a wide temperature range (25-65 degrees C). Digestion of lambda and psi X174 DNAs with BliI produced banding patterns identical to those seen with HaeIII. Therefore, BliI and HaeIII endonculeases are isoschizomers. PMID- 3032686 TI - Bromoacetylalprenololmenthane binding to beta-receptors modulates the rate of hormone-induced internalization but not desensitization in S49 cells. AB - Bromoacetylalprenololmenthane was found to inhibit hormone-induced beta adrenergic receptor internalization in a dose-dependent fashion in S49 lymphoma cells, besides its known ability to bind to beta-receptors irreversibly. This new found property of BAAM+ was taken advantage of in studying whether receptor internalization is a necessary step in the desensitization of adenylate cyclase. BAAM-treated cells showed functional desensitization even when receptor internalization had been blocked substantially by 50-65%. This finding suggests that receptor internalization is not directly involved in desensitization. PMID- 3032687 TI - The activation of protein kinase C by the polyphosphoinositides phosphatidylinositol 4,5-diphosphate and phosphatidylinositol 4-monophosphate. AB - Protein kinase C(PKC) is a Ca2+- and phospholipid-dependent protein kinase which can be activated by diacylglycerol, a product of polyphosphoinositide hydrolysis. In this report, we show that the polyphosphoinositides L-alpha phosphatidylinositol 4-monophosphate (PI 4P) and L-alpha-phosphatidylinositol 4,5 diphosphate (PI 4.5DP) can serve as phospholipid cofactors of isolated rat brain PKC. The order of potency of the phosphoinositides in the activation of PKC, PI greater than PI 4P greater than PI 4,5DP, shows a negative correlation with the degree of acidity of the phospholipid head group, whether 1 mM Ca2+ or 200 nM TPA is present in the reaction assay mixture. Although the polyphosphoinositides are by themselves weaker activators of PKC than PI, small amounts of PI 4,5DP cause a two-fold enhancement of PKC in the presence of Ca2+ and PI. While the endogenous phospholipid cofactors of PKC remain to be identified, these results suggest that the small amounts of polyphosphoinositides which are present in cell membranes may play a direct role in the activation of PKC in vivo, by serving as phospholipid cofactors of the enzyme. PMID- 3032688 TI - cAMP reduces the affinity of Ca2+-triggered secretion in platelets. AB - Prostacyclin and other related compounds known to increase intracellular cAMP levels inhibit platelet responses. The mechanisms involved are only partially known, especially those concerning the complex relations between Ca2+ and cAMP as opposite intracellular mediators. Here, we have investigated aggregation and secretion in quin2-loaded platelets under conditions in which Ca2+ and cAMP are the only intracellular mediators. Our results show that cAMP inhibits aggregation and secretion in ionophore-treated cell without modifying their intracellular Ca2+ levels. This result suggests that the inhibition takes place on some intracellular target for Ca2+. PMID- 3032689 TI - Fast abortive initiation of uvrA promoter in a supercoiled plasmid studied by stopped-flow techniques. AB - In order to follow the fast kinetics of abortive initiation (lag time from 1 ms to 10 s), we have built a stopped-flow apparatus equipped for fluorescence detection. The small volume used for each assay (35 microliters), and the short dead time (approximately 0.5 ms) are the essential advantages of this apparatus. Supercoiling of DNA affects considerably the initiation of transcription from the uvrA promoter. It decreases the lag time due to the isomerisation process 3-fold. Nevertheless, it does not change significantly the product KBk2, which is indicative of promoter strength and shows that uvrA is an 'association-limited' promoter. The presence of the LexA repressor increases the lag time considerably. At least for small RNA polymerase concentrations this increase is stronger for supercoiled than for linearized DNA. PMID- 3032690 TI - Oxygen-derived radicals: a link between reperfusion injury and inflammation. AB - Oxygen-derived free radicals (superoxide and hydroxyl) and related species (hydrogen peroxide and hypohalous acids) have well-defined roles in the inflammatory process. Their actions include the killing of microorganisms as well as participation in cell-to-cell communication among phagocytes via the activation of a superoxide-dependent chemoattractant. The active oxygen species also have roles in postischemic injury brought about by the conversion during ischemia of the enzyme xanthine dehydrogenase (EC 1.1.1.204) to the radical producing xanthine oxidase (EC 1.1.3.22). Although the enzymes responsible for producing superoxide in inflammation and ischemia are quite distinct, and are triggered by very different events, there are points of interplay in the two mechanisms whereby an ischemia/reperfusion-induced injury would lead to inflammation, and conversely whereby inflammation could lead to impairment of the circulation and hence to ischemic injury. PMID- 3032692 TI - Utility of the 24-hour delay hysterosalpingogram film. AB - Controversy exists regarding the ideal contrast media for hysterosalpingography. A unique property of oil-base contrast media is the availability of a 24-hour delay radiograph for further assessment of tubal patency and adhesions. A review was undertaken of the delay films in 131 cases performed by use of oil-base contrast media with subsequent surgical confirmation of pelvic findings. A 97% predictive accuracy was achieved with regard to distal obstruction and a 79% accuracy with regard to pelvic adhesions. Objective criteria for the evaluation of 24-hour delay hysterogram films were developed and are illustrated. PMID- 3032691 TI - Opioid regulation of luteinizing hormone in amenorrheic patients after therapy for induction of ovulation. AB - This study evaluated the activity of central opiate receptors modulating luteinizing hormone (LH) secretion before and during treatment with human menopausal gonadotropin (n = 8) or purified human urinary follicle-stimulating hormone (n = 6) in 14 patients with hypogonadotropic hypogonadism (n = 6) or secondary amenorrhea (n = 8). LH response to saline infusion and naloxone administration (4 mg intravenously) was assessed. As control, 6 normal ovulating women were studied. Before therapy, all amenorrheic patients showed no LH increase after naloxone injection. Gonadotropin treatment restored the naloxone induced LH response at preovulatory and midluteal phases in ovulating patients with secondary amenorrhea. The same response was present in spontaneously ovulating women but was absent in the hypogonadotropic hypogonad patients, despite the gonadotropin therapy's efficiency. In conclusion, when the alteration of gonadotropin-releasing hormone synthesis and/or release is reversible, the opioid system actively participates in the regulation of the hypothalamus pituitary-gonadal axis. PMID- 3032693 TI - Male infertility due to congenital adrenal hyperplasia: testicular biopsy findings, hormonal evaluation, and therapeutic results in three patients. AB - Testicular biopsy in three infertile male patients with congenital adrenal hyperplasia showed different grades of germinal epithelium maturation, and no Leydig cells in the interstitium. In two patients with severe germinal cell hypoplasia, very low serum gonadotropins showed no response to luteinizing hormone-releasing hormone stimulation and responded to long-term clomiphene citrate administration. It is hypothesized that an increase of hypothalamic estrogen production because of local androstenedione aromatization might be responsible for the luteinizing hormone-releasing hormone block observed in these patients. All patients were submitted to adrenal suppressive therapy, and in two cases a return of the fertility status was obtained. PMID- 3032694 TI - Hyperprolactinemia associated with clinically silent adenomas: endocrinologic and pathologic studies; a report of two cases. AB - Pituitary adenomas containing adrenocorticotropic hormone (ACTH) in one case, and ACTH, beta-lipotropin, and beta-endorphin in the other, were demonstrated in two patients who had amenorrhea-galactorrhea and hyperprolactinemia with no manifestation of Cushing's disease. Neither adenoma contained prolactin (PRL). Initial bromocriptine therapy resulted in cessation of amenorrhea-galactorrhea and normalization of PRL levels. However, there was radiologic evidence of tumor enlargement in both patients. After pituitary adenomectomy, the two patients resumed regular menses and normal PRL dynamics. These patients illustrate the need for bromocriptine therapy for possible enlargement of their pituitary adenomas. The diagnosis of silent corticotroph adenoma should be kept in mind. PMID- 3032695 TI - [Interneuronal relations in a dissociated sympathetic ganglion culture]. AB - Structural and functional relations between sympathetic neurons in the long-term culture of a dissociated autonomic ganglion were studied with light and electron microscopic (scanning microscopy) and electrophysiological microelectrode techniques. Different patterns of functional interneuronal relations depending on the ultrastructural organization and features of unification of the cells in networks, are discussed. PMID- 3032696 TI - [Effect of continuous illumination and disruption of monoaminergic innervation on the hypothalamus of the female rat]. AB - Hypothalamic suprachiasmatic nucleus (Schn) and Arquate nucleus (AN) were studied in female rats kept under constant illumination (rats with persistent estrus) or under light-dark cyclic illumination (normal estrus cycle). To destroy monoaminergic terminals, some rats from each group were injected with neurotoxic agents into the lateral cerebral ventricle. 6-hydroxydophamine which disturbs catecholaminergic terminals induced activation of the Schn and AN in rats with persistent estrus only. Destruction of serotoninergic terminals with 5,6 hydroxytryptophan induced activation of the Schn in rats with normal sex cycle and in those with a persistent estrus; the AN was also activated but only in rats with normal estrous cycle. As reported earlier, destruction of serotoninergic terminals withstands the decrease of plasma LH under constant illumination although persistent estrus still lasts. It seems there are different parallel ways to disturb normal sex cycle under constant light illumination. PMID- 3032698 TI - Two exons specific for the myb proto-oncogene found upstream from the avian myeloblastosis virus-transduced myb sequences. AB - A partial restriction map of cloned 5.42-kb chicken DNA (clone P542, Perbal et al. 1983), covering a portion of the c-myb locus, is presented. The 5' end of the v-myb gene (approximately 0.5 kb) is located at the 3' end of P542 DNA, the remainder are the cellular sequences not transduced by avian myeloblastosis virus. Two non-contiguous DNA segments were detected within these cellular sequences which code for the 5' end of c-myb mRNA. These two exons, designated e1 and e2, are separated by a approximately equal to 1.5-kb non-coding region. Both of them are transcribed into 0.4 kb located near the 5' end of c-myb mRNA. The second exon e2 (approximately equal to 0.2 kb) is flanked at its 3' end by a short non-coding region within which virus-cell recombination took place. The possible presence of a portion of this intron in the 2.1-kb v-myb mRNA is discussed. PMID- 3032697 TI - [Intracellular study of the formation of new synapses and collateral sprouting of red nucleus neurons after destruction of the cerebellar nucleus interpositus in the adult cat]. AB - Responses of red nucleus neurons to stimulation of ipsilateral cerebellar nucleus interpositus were studied in cats after lesion of contralateral cerebellar nucleus interpositus within the period ranging from two weeks to one year and seven weeks. Recording of fast and slow mono- and polysynaptic EPSPs showed collateral sprouting of axons of interposito-rubral neurons to occur in various parts of soma-dendritic membrane of rubro-spinal neurons. Stimulation of ipsilateral cerebellar nucleus interpositus evoked antidromic activation in many red nucleus neurons, suggesting the sprouting of axon collaterals from rubro spinal neurons onto the neurons of ipsilateral cerebellar n. interpositus projecting to the red nucleus. PMID- 3032699 TI - Transfer of Drosophila melanogaster transponsable genetic element mdg-4 into plant cells. AB - The copia-like element mdg-4, as a component of the Drosophila genomic fragment Dm111, was cloned into the vector pIB16. The chimaeric plasmid pIB16 [Dm111] was used to transform tobacco cells as a cointegrate with pTiC58 (Agrobacterium tumefaciens), pTiC58::pIB16 [Dm111]. The growth properties of primary crown-gall tumours were followed, and the DNA of one Nicotiana tabacum transformant was further analysed. The DNA/DNA molecular hybridization of digests of genomic DNA with 32P-labelled pDm111 probe demonstrated full-length insertion of Dm111 into N. tabacum genome. We were not able to detect any common sequence with Dm111 in untransformed tobacco cells. PMID- 3032700 TI - [Influence of endogenous renin-angiotensin system on plasma aldosterone responsiveness to metoclopramide]. AB - Hyperresponsiveness of plasma aldosterone to metoclopramide (MCP) was found in the patients with secondary aldosteronism. To understand the mechanisms of aldosterone reactivity to MCP, we examined the changes of its responsiveness to MCP and ACTH in various states of endogenous renin-angiotensin system which were induced by administrating two antihypertensive agents to patients with essential hypertension. Aldosterone responsiveness to both stimulants were accentuated during diuretic antihypertensive therapy in comparison with those before therapy. After adding converting enzyme blocker to diuretic agent, aldosterone reactivity to MCP decreased. These results indicates that aldosterone responsiveness to MCP which was thought to be controlled by changes of endogenous dopamine may be also influenced by the state of endogenous renin-angiotensin system. PMID- 3032701 TI - Improved bridge aesthetics: alveolar ridge augmentation. PMID- 3032702 TI - Cold reacting antilymphocyte antibodies in type I (insulin dependent) diabetes. AB - Non HLA antilymphocyte antibodies have been detected in numerous auto-immune illnesses and notably in type I (insulin dependent) diabetes. In order to ascertain their role in this illness, the actual frequency of this phenomenon has been estimated. The privileged cellular target has been determined. Finally correlations with the other immunogenetic markers of diabetes have been investigated. Non HLA antilymphocyte antibodies are frequent, above all at the beginning of the illness (32.4% before 6 months, 10% after 5 years). The cellular target is mainly composed of B lymphocytes. B lymphocyte enrichment of cellular suspensions leads to better method sensitivity (70.3% positive reactions at the beginning of the illness). No correlation was found with presence of anti islet cell antibodies or anti coxsackie B virus antibodies (IgM). Association with the HLA DR3 phenotype is relatively frequent but does not attain statistical significance. PMID- 3032703 TI - Effect of Li+ and Ba2+ on the electrocyte membrane-bound (Na+ + K+)-ATPase. AB - Membrane-bound (Na+ + K+)-ATPase activity from the non-innervated and innervated faces of Electrophorus electricus (L.) electric organ, obtained by differential centrifugation, was measured using AChE as an enzyme marker for membranes derived from the post-synaptic area (fraction P3) of the electrocyte. The effect of Li+ and Ba2+ on (Na+ + K+)-ATPase activity of the two membrane fractions (P2 and P3) was analysed with respect to K+ and Mg2+ ions, after the I50 estimation. The kinetics of the reactions with these cations were investigated showing that Li+ inhibits P2 uncompetitively and for P3 presented a mixed type inhibition. Ba2+ behaved as an hyperbolic mixed type inhibitor for P2 and a linear mixed type inhibitor for P3 fraction. PMID- 3032705 TI - An early effect of estradiol at hepatic level, previous to its protein synthesis activation. AB - Liver glycogen phosphorylase activity is increased before protein synthesis activation by estradiol. This effect is not blocked by antibiotics (actinomycin D and cycloheximide) inhibitor of protein synthesis. At times very similar to those of phosphorylase activation, cAMP levels are not enhanced, as would be expected, but slightly depleted. At similar times, cGMP levels are dramatically increased. PMID- 3032704 TI - Calmodulin in porcine malignant hyperpyrexia. AB - The anaesthetic complication malignant hyperpyrexia (MH) is due to an elevation of the myoplasmic Ca2+ concentration. Examination of calmodulin isolated from MH susceptible swine suggests that the disorder in calcium regulation in MH is not due to an abnormality in calmodulin. PMID- 3032706 TI - Phosphodiesterase from the venom of Crotalus ruber ruber. AB - Phosphodiesterase was isolated from the venom of Crotalus ruber ruber from the U.S.A. using the gel filtration on a Sephadex G-75 column, followed by anion or cation exchange chromatography. Phosphodiesterase was homogeneous as established by a single band on acrylamide gel electrophoresis and isoelectric focusing electrophoresis. Phosphodiesterase activity was inhibited by ethylenediamine tetraacetic acid (EDTA), o-phenanthroline, thioglycolic acid or p chloromercuribenzoate (PCMB), but not by soybean trypsin inhibitor (SBTI) or benzamidine. The molecular weight of this enzyme was determined to be approx. 98,000 and the isoelectric point was found to be pH 10.5 by isoelectric focusing with carrier ampholyte. This enzyme contained 1.04 mol zinc per mol. The Michaelis constant (Km) of this enzyme for p-nitrophenyl thymidine-5'-phosphate and inhibition constant (Ki) for PCMB were found to be 8.3 X 10(-3) and 1.2 X 10( 2) M, respectively. PMID- 3032707 TI - Cellular localization and developmental changes of deoxyribonucleoside-activated nucleotidase in the rat testis. AB - Specific deoxyribonucleoside-activated nucleotidase (DAN) activity showed a rapid decline during the exponential increase in testis weight between 25 and 35 days of age. Specific DAN activity in Sertoli cells was dependent on the amount of cytosol in the enzyme assay. At optimal cytosol concentration the measured value was 50 units/mg of protein. Specific DAN activity in peritubular cells, primary spermatocytes and round spermatids was 13, 3.7 and 3.1 units/mg, respectively, and was independent of the cytosol concentration in the assay. PMID- 3032708 TI - Oestrogens inhibit steroid production by dispersed porcine thecal cells. AB - An intra-ovarian role for oestrogens in the control of steroid production was investigated using dispersed thecal cells obtained from porcine follicles. Thecal cells were incubated for 14 h at 37 degrees C and the media subsequently assayed for androstenedione, progesterone and cyclic AMP. LH caused a dose-dependent stimulation of both steroids and the addition of oestradiol at doses of 10 ng-10 micrograms/ml significantly (P less than 0.01) inhibited both basal and LH stimulated steroid production from doses of 500 ng/ml and upwards. Of other oestrogens investigated, oestrone and oestriol were somewhat less potent than oestradiol in inhibiting steroid synthesis, whereas the synthetic oestrogen diethylstilbestrol (DES) was more potent. The presence of oestradiol at doses of 10 ng-10 micrograms/ml had no significant effect (P less than 0.05) on either basal or LH-stimulated cAMP suggesting that the oestradiol inhibition does not involve inhibition of LH receptor-linked adenylate cyclase. These results demonstrate that physiological doses of oestrogen can act by local negative feedback to control the synthesis of its own precursor and thus regulate intrafollicular steroidogenesis. PMID- 3032709 TI - Modulation of Leydig cell functions by culture with Sertoli cells or with Sertoli cell-conditioned medium: effect of insulin, somatomedin-C and FSH. AB - The potential regulatory action of Sertoli cells on Leydig cell functions has been investigated by using a coculture system of Leydig and Sertoli cells obtained from immature pig or by culturing Leydig cells with Sertoli cell conditioned medium (SCCM). Coculture of Leydig and Sertoli cells for 48 h in the absence of insulin or somatomedin-C (Sm-C), produced a small but significant increase in both hCG receptors and hCG-induced testosterone production. Addition to the medium of pFSH (100 ng/ml), insulin (5 micrograms/ml) or somatomedin-C (50 ng/ml) produced a marked increase in these two parameters of Leydig cell function. A further significant increase was observed when pFSH was associated with insulin or Sm-C. In contrast, coculture of Leydig cells with aortic endothelial cells decreased both the hCG receptor number and the hCG responsiveness. SCCM stimulated Leydig cell testosterone production following a 4 h incubation. The stimulation depended upon the amount of SCCM used and the conditions in which Sertoli cells were cultured. The most active was the medium from cells cultured in the presence of pFSH and insulin at high concentrations. Since pig Sertoli cells have specific somatomedin-C receptors, but not insulin receptors, it is likely that the effect of micromolar concentrations of insulin are exerted through Sm-C receptors. In addition, SCCM produced a long-term effect after 48 h incubation. SCCM from cells cultured in the absence of insulin and pFSH inhibited both the hCG receptor number and hCG responsiveness. A similar inhibition was observed with SCCM medium from cells cultured without insulin but with pFSH.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3032710 TI - Relationship between renin mRNA and renin secretion in adrenalectomized, salt depleted, or converting enzyme inhibitor-treated rats. AB - Renin secretion was stimulated in rats by adrenalectomy (ADX), by treatment with the converting enzyme inhibitor enalapril (MK 421), or by feeding a low salt diet plus furosemide treatment (FUR). Relative amounts of renin mRNA were measured by densitometric Northern blot analysis using a 32P-labeled 700 bp mouse submaxillary gland cDNA fragment as hybridization probe. Renin secretion was measured in isolated kidneys taken from pretreated rats. Treatment resulted in a 4.4-, 7- and 12.7-fold increase in total renin mRNA in kidneys of ADX, MK 421 and FUR rats, respectively. Plasma renin levels increased 5-, 9- and 12-fold in ADX, MK 421 and FUR rats, whereas ex vivo secretion rates from the isolated kidney were 4.7-, 0.5- and 4.2-fold, respectively, that of controls. Thus, a close relationship between increases in renin secretion in vivo (reflected by plasma renin concentration) and renin mRNA exists, whereas secretion rates in the isolated perfused kidney ex vivo change in a non-proportional fashion. PMID- 3032711 TI - Binding of (+)-PN 200-110 to rat pituitaries and to normal and adenomatous human pituitaries. AB - Endocrine cells possess voltage-sensitive Ca2+ channels involved in the modulation of hormonal secretion. Using the dihydropyridine, (+)-PN 200-110, we have investigated the binding characteristics of this ligand to pituitary membrane Ca2+ channels from normal rat, normal and adenomatous human pituitaries. [3H]PN 200-110 binds specifically to rat pituitary membranes to one class of sites (Kd = 0.41 +/- 0.10 mM; Bmax = 39 +/- 1.3 fmol/mg protein). At 37 degrees C, equilibrium is reached in 45 min and half-life of the binding is 13 min. No significant changes were observed for either the Kd or Bmax values between normal rat and human pituitaries or between the different types of adenomas (GH- and PRL secreting and non-secreting). As the secretory activity of the pituitary adenomas, involving Ca2+ mobilization, varies from one adenoma to another, our results could indicate that, if there is a modified regulation of Ca2+ entry in the adenomas, it may not be related to a varying number of calcium channels, at least the channels labeled by the dihydropyridine (+)-PN 200-110. PMID- 3032712 TI - A quantitative study of the developmental expression of nerve growth factor (NGF) receptor in rats. AB - The developmental expression of nerve growth factor (NGF) receptor was quantitated in either homogenates or plasma membrane-enriched preparations from whole rat embryos or from isolated tissues. The assay involved crosslinking 125I NGF to receptors followed by immunoprecipitation with a monoclonal antibody to rat NGF receptor. In some cases, the pellet was resuspended and subjected to a sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) autoradiographic analysis. The NGF receptor was found in whole embryo homogenates as early as embryonic Day 10 (E10) (earliest age examined). The NGF receptor content in whole embryos per milligram protein increased about 3-fold from E11 to E18 and decreased slightly at E20. SDS-PAGE autoradiography showed that the molecular weights of 125I-NGF-bound receptors did not vary with age. The NGF receptor content in sciatic nerve homogenates decreased 23-fold from newborn to adulthood. The change of NGF receptor level in hindleg muscle had a profile similar to that seen in sciatic nerve. The NGF receptor content in superior cervical ganglion (SCG) or dorsal root ganglion (DRG) homogenate preparations was expressed in two ways. On a per milligram protein basis, in SCG, the receptor density was decreased slightly from E20 to adulthood; in DRG, it was relatively constant from E15 through postnatal Day 0 (PND-0) and then dropped 6.7-fold in adults. On a per ganglion basis, in SCG, it increased 4.4-fold from E20 to adult; in DRG, it increased 9-fold from E15 to PND-0 and then stayed constant through adulthood. In brain membrane preparations, the NGF receptor level decreased 11 fold from E15 to adulthood. In spinal cord membrane preparations, it decreased 7 fold from E18 to adulthood. Levels of receptor in cord were always greater than in brain. These data suggest that alterations in the NGF receptor density may have a role in changes in tissue responsiveness to NGF during development. PMID- 3032713 TI - A rise in cytosolic calcium is not necessary for maturation of Xenopus laevis oocytes. AB - Cytoplasmic free calcium levels during progesterone-induced meiotic maturation in Xenopus laevis oocytes were measured using the photoprotein aequorin. The resting level of [Ca2+]i was 92.6 +/- 30 nM. No significant changes were observed after progesterone addition, although a large pulse of [Ca2+]i was observed upon activation of matured oocytes. These findings are discussed in terms of the role of calcium in maturation and it is concluded that calcium is not the second messenger for progesterone. This conclusion is further supported by the finding that 100 microM TMB-8, a blocker of intracellular calcium release, had no effect on progesterone-induced maturation. PMID- 3032714 TI - Evidence for the involvement of PI-signaling and diacylglycerol second messengers in the initiation of metamorphosis in the hydroid Hydractinia echinata Fleming. PMID- 3032715 TI - Insulin-receptor kinase activity of adipose tissue from morbidly obese humans with and without NIDDM. AB - We have determined glucose transport, insulin binding, and insulin-receptor kinase activity in adipose tissue from morbidly obese patients with and without non-insulin-dependent diabetes mellitus (NIDDM). The insulin sensitivity and responsiveness of glucose transport in freshly isolated adipocytes were significantly reduced in NIDDM subjects compared with nondiabetics. This was due in part to decreased insulin binding in adipocytes. Reduced specific 125I-labeled insulin binding was also observed in crude detergent extracts and partially purified insulin receptors from adipose tissue. In addition, the basal and insulin-stimulated tyrosine-specific protein kinase activity per milligram of protein was significantly decreased in NIDDM patients compared with nondiabetics. The differences between maximally insulin-stimulated and basal kinase activities expressed by insulin-binding activity were also significantly reduced in NIDDM subjects. We conclude that insulin resistance in morbidly obese patients with NIDDM is due to both insulin-binding and postbinding defects. One of the postbinding defects in NIDDM appears to be impaired insulin-receptor kinase activity of fat tissue. PMID- 3032716 TI - Direct stimulation of Na+-K+-ATPase and its glucosylated derivative by aldose reductase inhibitor. AB - In the presence of 10(-8) M concentrations of the aldose reductase inhibitor AL 1576, there is a 20-30% increase in the rate of hydrolysis of near-saturating concentrations of ATP by bovine renal Na+-K+-ATPase. When bovine renal Na+-K+ ATPase is reacted with glucose 6-phosphate in the presence of 10(-8) M concentrations of AL 1576 or 10(-6) M concentrations of a second aldose reductase inhibitor, sorbinil, glucosylation occurs. Whereas sorbinil has no effect on ATP hydrolysis by the glucosylated Na+-K+-ATPase, 10(-8) M AL 1576 causes a shift in the kinetics of hydrolysis of ATP from substrate inhibition to normal substrate activation. The aldose reductase inhibitors interact with the enzyme at the low affinity ATP-binding site. PMID- 3032717 TI - Galactose neuropathy. Structural changes evaluated by nuclear magnetic resonance spectroscopy. AB - We examined the hypothesis that the polyol accumulation resulting from chronic galactose supplementation in the diet produces endoneurial edema that can be prevented by inhibition of aldose reductase. We explored the potential of nuclear magnetic resonance (NMR) spectroscopy to quantitate and characterize the water accumulation in the sciatic nerve in this "galactose neuropathy." The data demonstrate a 16% increase in gravimetrically determined total water content of nerve in the galactose-fed rat after 8 mo of this diet and a 50% increase in the T1 relaxation time for nerve water as determined by NMR spectroscopy. Prolongation of the T1 relaxation time reflects increased rotation of water in a magnetic field, consistent with an extracellular site of the additional water. Simultaneous feeding of sorbinil to inhibit aldose reductase resulted in normalization of both total nerve water and of the prolongation of T1 relaxation time. These data define the NMR-spectroscopic state of endoneurial edema in the galactose-fed rat and suggest specific application to the investigation of the role of aldose reductase in human diabetic neuropathy. PMID- 3032718 TI - Increased responsiveness to glucoregulatory effect of opiates in obese-diabetic ob/ob mice. AB - Plasma glucose and insulin responses to opiate receptor stimulation and antagonism were determined in 12-14 week old lean and obese-diabetic Aston ob/ob mice. The opiate receptor antagonist naloxone (1 mg/kg intraperitoneally) rapidly and transiently raised glucose and suppressed insulin concentrations in lean mice, and produced qualitatively similar but more protracted response in ob/ob mice. Selective stimulation of mu- and delta-opiate receptors using the enkephalin analogues Tyr-D-Ala-Gly-MePhe-NH(CH2)2OH (1 mg/kg, intraperitoneally) and Tyr-D-Ala-Gly-Phe-D-Leu (10 mg/kg intraperitoneally) respectively, rapidly and transiently increased glucose and insulin concentrations in lean and ob/ob mice. The ob/ob mice exhibited greater glucose and insulin responses to these analogues. The results provide evidence that endogenous opiates participate in the regulation of glucose and insulin homeostasis, and suggest that increased responsiveness to mu- and delta-opiate receptor stimulation may contribute to the hyperglycaemia and hyperinsulinaemia of obese-diabetic mice. PMID- 3032720 TI - Altered cyclic-AMP receptor activity and morphogenesis in a chemosensory mutant of Dictyostelium discoideum. AB - The functional properties of the cell-surface cyclic-AMP receptor that controls chemotaxis were found to be altered in an aggregation mutant of Dictyostelium discoideum. The mutant aggregated without stream formation and had a tenfold increased cell-density requirement for the initiation of aggregation. After aggregation, mounds formed multiple tips and subsequently subdivided to give multiple fruits that were small and abnormally proportioned. Cyclic-AMP-induced light-scattering changes in cell suspensions indicated that the mutant had a diminished response to external cyclic-AMP signals. Associated with these altered functional responses was a physical change in the cyclic-AMP sensory system. Cyclic-AMP-binding studies showed that the parent had two classes of cyclic-AMP binding sites, i.e., Kd = 32 and 110 nM. In contrast, the mutant had two- to threefold or more high-affinity sites (Kd = 25 nM) and altered low-affinity sites (Kd less than 3 microM). These results indicate that both affinity classes of binding site are independently mutable. This observation suggests that the two affinity classes can be interconverted by mutation, or the mutation alters a single molecular species and its equilibrium between binding sites with different affinities for cyclic AMP, as postulated in receptor cycling models. PMID- 3032719 TI - Catecholamines and tumour promoting phorbolesters inhibit insulin receptor kinase and induce insulin resistance in isolated human adipocytes. AB - The effect of the catecholamine isoprenaline (10(-5) mol/l) and of the tumour promoting phorbolester tetradecanoyl-beta-phorbol acetate (10(-9) mol/l) on insulin stimulated 3-O-methyl-glucose transport was studied in freshly isolated human adipocytes. Both substances reduced the maximal responsiveness of the glucose transport system to insulin by approximately 50%. To test if this is caused by inhibition of the insulin receptor kinase the receptor from phorbolester and isoprenaline treated cells was solubilized, partially purified and its kinase activity studied in vitro. Insulin stimulated 32P-incorporation into the beta-subunit of the insulin receptor of phorbolester or isoprenaline treated cells was reduced to 20-60% of the values found with receptor from control cells at insulin concentrations between 10(-10) mol/l and 10(-7) mol/l. This inhibition of kinase activity of receptor from phorbolester and isoprenaline treated cells was observed at nonsaturating adenosine triphosphate levels (5 mumol/l), and it could be overcome with higher concentrations of gamma-32P adenosine triphosphate in the phosphorylation assay. A Lineweaver Burk analysis of the insulin stimulated receptor phosphorylation revealed that the Michaelis constant for adenosine triphosphate of the receptor kinase from phorbolester and isoprenaline treated cells was increased to greater than 100 mumol/l compared with less than 50 mumol/l for receptor from control cells. We conclude from the data that catecholamine and phorbolester treatment of human adipocytes modulates the kinase activity of the insulin receptor by increasing its Michaelis constant for adenosine-triphosphate, and propose that this modulation of receptor kinase is a mechanism that can contribute to the pathogenesis of insulin resistance in human fat cells. PMID- 3032721 TI - Chondrogenesis in mouse limb buds in vitro: effects of dibutyryl cyclic AMP treatment. AB - We studied the effects of dibutyryl cyclic AMP (dbcAMP) on mouse limb-bud chondrogenesis at three stages of embryonic development. After 24 h of culture, limb buds with or without a covering of ectoderm were treated with 1 mM dbcAMP for 48 h and were then compared with untreated cultured limb buds. Treatment with dbcAMP enhanced cartilaginous differentiation in organ cultures of stage-17 and 19 (according to Theiler's) limb buds, although the presence of ectoderm reduced the level of dbcAMP stimulation. By stage 20, treatment with dbcAMP irreversibly inhibited cartilaginous differentiation. These results suggest that the responsiveness of mesenchymal limb-bud cells to dbcAMP is stage related. The results of histological studies as well as of analyses of DNA content and sulphated glycosaminoglycan accumulation supported the hypothesis that dbcAMP treatment induces recruitment of initially non-chondrogenic cells whose commitment explains the enhancement of cartilaginous differentiation. Limb-bud competence for chondrogenesis throughout the three developmental stages studied is also discussed. PMID- 3032722 TI - Retinoic-acid-induced differentiation of HL-60 cells is associated with biphasic activation of the Na+-K+ pump. AB - The human promyelocytic leukemia cell line, HL-60, can be induced to differentiate into granulocyte-like cells when cultured in the presence of 10(-6) M retinoic acid (RA) for several days. Following the addition of RA two kinds of changes occur. First, there are early changes that comprise an increase in the intracellular concentration of sodium ions [Na]i, which reaches its maximum after 6 h, and an increase in the activity of the Na+-pump, which is reflected by an ouabain-sensitive K+ influx that peaks at 8 h (170% of the control value) and that occurs without any change in the number of pump molecules, as measured by the binding of 3H-ouabain. Second, beginning after 12 h of culture with RA, a decrease in the number of ouabain-binding sites occurs, this being accompanied by an increase in the number of K+ ions actively transported by each site. The effect of modulation of Na+-pump activity on the RA-induced differentiation of HL 60 cells was studied using low, noncytotoxic concentrations of ouabain which, although alone having no differentiating effect, accelerated and potentiated the effect of RA on differentiation. When added in combination, these drugs induced rapid stimulation of the Na+-pump, which reached its peak after 2 h. These results indicate that a concomitant increase in the level of [Na+]i and in the activity of the Na+-pump constitute primary events in the interaction between RA and HL-60 cells, and that cation fluxes may play a role in the initiation of the process of differentiation. PMID- 3032723 TI - [Hepatocellular carcinoma invading the right atrium: clinical, echographic and angiographic study]. AB - The authors report the case of a 54 year old woman suffering from hepatocellular carcinoma with tumor growth into right hepatic vein, inferior vena cava and right atrium. On cardiac examination, a pansystolic bruit and a diastolic rumble were audible at the tricuspid focus. Diagnosis was confirmed by inferior vena cavography and two-dimensional echocardiography, which demonstrated a large mobile mass in the right atrium moving to and fro through the tricuspid valve. This case report emphasizes the value of routine cardiac examination during the course of hepatocellular carcinoma. PMID- 3032724 TI - [Acute hepatitis after taking enalapril maleate (Renitec)]. PMID- 3032725 TI - Hepatitis B virus DNA in human hepatocellular carcinoma: is the integration of hepatitis B virus DNA really carcinogenic? AB - Hepatitis B virus-specific DNA sequences (HBVDNA) in the liver were examined in 19 patients with hepatocellular carcinoma (HCC), 5 patients with liver cirrhosis (LC) and without HCC, 3 patients with chronic hepatitis (CH), 2 patients with metastatic liver cancer (MLC), 1 patient with primary biliary cirrhosis (PBC) and 4 patients with normal liver (NL) by the Southern blot hybridization procedure. Integration of HBVDNA was found in all 4 HCC patients with serum HBsAg, of whom one patient had HBVDNA only in the non-tumor (cirrhotic) region. Integration of HBVDNA was also detected in 4 of 8 HCC patients without serum HBsAg but with serum HBV-related antibodies, and in 2 HBsAg-positive patients of 5 LC patients. All 3 HBsAg-positive CH patients had only extrachromosomal HBVDNA. No HBVDNA was detected in the other 21 patients. Although integration of HBVDNA was observed in HBsAg-positive HCC patients with a higher frequency, integrated HBVDNA could also be detected in non-tumor regions of HCC patients and cirrhotic livers without HCC. It was concluded from these observations that integration of HBVDNA was frequently associated with HCC but might not have a direct causal effect on hepatocarcinogenesis even in HBV carriers. PMID- 3032726 TI - Hepatic conversion of 1-(tetrahydro-2-furanyl)-5-fluorouracil into 5-fluorouracil in patients with hepatocellular carcinoma. AB - The pharmacokinetics of 1-(tetrahydro-2-furanyl)-5-fluorouracil (FT) and its conversion into 5-fluorouracil (FUra) in liver tissue were studied in ten patients with hepatocellular carcinoma (HCC). The plasma concentration of FT after its intravenous injection (dosage: 800 mg) was computerfitted to a bi exponential function (C = Ae-alpha t + Be-beta t), indicating a two-compartment disposition. The pharmacokinetic parameters did not significantly differ between the five patients with, and the five without cirrhosis of the liver. The plasma concentrations of FUra likewise showed no significant difference between the two groups. The rates of FT degradation in the liver tissue homogenate were similar for four of the patients with cirrhosis (0.10 +/- 0.05 mumol/g liver protein/30 min) and four of those without it (0.13 +/- 0.05). The rates of cytochrome P-450 dependent FUra formation in the microsomal fraction of liver tissue from two patients (1.1 and 1.3 nmol/mg microsomal protein/30 min) were dramatically reduced to less than half of those of two control subjects (2.4 and 2.7). The estimated rates of FUra formation in the soluble fraction (105,000 X g supernatant fraction) from the two patients (0.1 and 0.13 nmol/mg protein/30 min) were almost identical to those from the controls (0.12 and 0.14), suggesting that the rate in the soluble fraction from HCC patients may not be as strongly affected as the rate in the microsomal fraction.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3032727 TI - Gastric plasmacytoma with cylindrical crystalline inclusions. AB - An 81 year-old female case of extramedullary plasmacytoma of the stomach is reported. Immunohistochemically, the tumor cells contained IgM-lambda immunoglobulin. Electron micrographs of the tumor cells showed centrally located nuclei with nuclear inclusions and cytoplasm filled with crystalline inclusions in the rough endoplasmic reticulum. IgM-lambda producing gastric plasmacytoma with cylindrical inclusions is rare. PMID- 3032728 TI - Granular cell tumor of the ascending colon: a case report. AB - A granular cell tumor (granular cell myoblastoma) of the ascending colon in a 48 year old male is reported. The tumor was detected by barium enema study as a sessile polyp, and colonofiberscopy revealed submucosal tumor. It was removed by endoscopic polypectomy. Macroscopic examination showed the characteristic features of granular cell tumor. The avidin-biotin-peroxidase complex (ABC) method for detection of S-100 protein demonstrated that the cytoplasm of tumor cells and the pleomorphic nuclei were strongly stained with anti-S-100 protein serum, which supports the concept of the Schwann cell origin of granular cell tumor. PMID- 3032729 TI - Bile duct abnormalities in the acquired immune deficiency syndrome. AB - Pathological changes in the liver have been described in patients with acquired immune deficiency syndrome; however, lesions of the bile ducts have not been noted. We report 2 patients with intrahepatic bile duct abnormalities, one having sclerosing cholangitis of the large ducts. We discuss the possibility that these changes are secondary to opportunistic infection or are induced more directly by immunologic mechanisms. PMID- 3032730 TI - Thyroid hormone-induced changes in different ion-specific adenosine triphosphatase activities in liver of Singi fish Heteropneustes fossilis (Bloch). AB - A single injection of different doses of T3 (0.5, 5, 20, and 50 micrograms/g) to Singi fish caused an increase in Na+K+-ATPase activity in crude liver homogenate in a dose-dependent non-linear fashion on the 3rd d. Ca++- and Mg++-ATPase activity increased only with 20 and 50 micrograms/g of T3. Lowering the dose of T3 to 0.1 microgram and 0.25 microgram/g in a single injection had not effect on these enzyme activities. TETRAC (1, 2, and 4 micrograms/g) and TRIAC (2 and 4 micrograms/g) in a single injection enhanced the activities of Na+K+-ATPase, but Ca++- and Mg++-ATPase activities remained unchanged on the 3rd d. Immersion of Singi fish in thiourea-containing medium (1 mg/ml) for 30 d caused reduction in Na+K+-ATPase activity, but Ca++- and Mg++-ATPase activity remained unaltered. The reduced level of Na+K+-ATPase activity in the thiourea-treated hypothyroid fish was recovered and even brought above the control level by a single injection of T3 at the dose of 0.5 microgram/g. Differential sensitivity of various ion specific ATPases to T3 in liver of Singi fish is thus documented. PMID- 3032731 TI - Involvement of central alpha-pressor and beta-depressor adrenoceptors in the cardiovascular response to intracerebroventricular catecholamines in the rat. AB - Microinjections of catecholamines were performed into the lateral ventricle of anesthetized and unanesthetized rats and the blood pressure effects recorded prior to or after the administration of pharmacological antagonists. Injections of 20-100 micrograms normetanephrine (alpha-agonist) produced only pressor responses in both groups of animals. Injections of 10-20 micrograms of norepinephrine (preferentially an alpha-agonist) produced mainly pressor in awake and only depressor responses in anesthetized animals, whereas injections of 10-20 micrograms of epinephrine (preferentially a beta-agonist) produced only depressor responses in both groups of animals. Intracerebroventricular pretreatment with the beta-blocker propranolol (40-100 micrograms) blocked the depressor responses to the catecholamines or even reverted them into clear pressor response. Pretreatment with the alpha-blocker phentolamine (100 micrograms) reduced the pressor effects induced by the intraventricular injection of catecholamines. The existence of central alpha-pressor and beta-depressor mechanisms mediating the blood pressure responses to the intracerebroventricular administration of catecholamines is proposed. PMID- 3032732 TI - Effects of bunazosin on electrical responses of smooth muscle cells of the guinea pig mesenteric artery and vein to perivascular nerve stimulation and to noradrenaline. AB - The noradrenaline-induced depolarization of smooth muscle cell membrane was blocked by bunazosin in the mesenteric artery but not in the mesenteric vein. Bunazosin enhanced the excitatory junction potential (e.j.p.) evoked in the mesenteric artery but did not modulate the slow depolarization evoked in the mesenteric vein. Application of noradrenaline decreased the amplitude of e.j.p. enhanced by bunazosin but not by yohimbine. It was concluded that bunazosin is a highly selective alpha 1-adrenoceptor blocker in vascular tissues. PMID- 3032733 TI - TRH-induced antinociception: interaction with the opioid systems? AB - Mice were chronically treated with morphine or ethylketocyclazocine in order to induce a marked tolerance to their antinociceptive effect in the phenyl-p benzoquinone writhing test. TRH (2 mg kg-1 i.p.) significantly reduced the number of writhes in non-tolerant mice, but did not alter the response of morphine- or ethylketocyclazocine-tolerant mice. TRH did not modify the binding of [3H]naloxone in mouse brain either in vitro (TRH: 10(-10)-10(-4) M) or ex vivo (TRH: 1-40 mg kg-1 i.p.). There was no dose-dependent modification of the in vivo binding of [3H]lofentanil in any of the mouse brain areas studied after TRH (1-40 mg kg-1 i.p.). PMID- 3032734 TI - Noradrenaline inhibits vasoconstriction induced by electrical stimulation. AB - Exposure of the isolated rat tail artery to exogenous noradrenaline inhibited vasoconstriction induced by electrical field stimulation. Phenylephrine produced brief inhibition; guanfacine potentiated electrical stimulation. Sympathetic neurotransmission may be blunted by brief increases in circulating noradrenaline levels. PMID- 3032735 TI - Central receptor binding and cardiovascular effects of GABA analogues in the cat. AB - Several structural analogues of GABA were shown to be inhibitors of GABAA receptor binding in membranes from cat cerebral cortex. These compounds were 3 aminopropanesulfonic acid (APS; IC50 = 0.04 microM), imidazoleacetic acid (IMA; IC50 = 0.4 microM), morpholinopropanesulfonic acid (MOPS; IC50 = 1.6 microM), 5 phenylpyrrolepropionic acid (PPP; IC50 = 15 microM), aminoethanethiosulfonic acid (AETS; IC50 = 22 microM), 3-amino-3-phenylpropionic acid (APP; IC50 = 35 microM), meta-aminobenzoic acid (MABA; IC50 = 58 microM) and urocanic acid (UCA; IC50 = 354 microM). The IC50 value for GABA was 0.03 microM. GABA, PPP, AETS, MABA and UCA were previously shown to reduce arterial pressure in the cat after intracerebroventricular infusion. In the present study MOPS (ED50 = 0.26 nmol/kg), APS (ED50 = 4.7 nmol/kg), APP (ED50 = 49 nmol/kg), and IMA (ED50 = 350 nmol/kg) were also found to be able to decrease blood pressure when infused into the fourth ventricle. All nine compounds reduced blood pressure to the same extent, but in some cases their relative potencies (ED50 values) exhibited significant differences. When the IC50 values for receptor binding were plotted against the ED50 values for the cardiovascular effects, no significant correlation emerged. This lack of a correlation does not necessarily imply that the reductions in blood pressure elicited by the drugs are not related to an activation of central GABAA receptors. Instead, it highlights the difficulties that are sometimes encountered in attempting to obtain quantitative measurements after intracerebroventricular infusion. PMID- 3032736 TI - The failure of morphine to interact with serotonin on nucleus raphe magnus descending inhibition or morphine-induced suppression of cat spinal cord multireceptive neurones. AB - Morphine sulphate, administered in three cumulative doses (0.5, 1.0, and 2.0 mg/kg, i.v.) to alpha-chloralose anaesthetized cats, reduced the nociceptive activity of deep dorsal horn multireceptive neurones but failed to alter the descending nucleus raphe magnus (NRM) phasic inhibition of these neurones. Morphine was also administered to fluoxetine (6.0 mg/kg, i.v.) pretreated animals. Fluoxetine is a selective 5-hydroxytryptamine (5-HT) uptake blocker, which should enhance 5-HT synaptic transmission. In these animals, morphine suppressed the neuronal nociceptive activity to the same extent as seen with morphine alone and did not affect the NRM inhibition. These results do not support the notion that morphine activates a descending serotonergic inhibition from the NRM or that serotonin mediates morphine inhibition of spinal nociceptive transmission in cats. PMID- 3032737 TI - Effects of cytotoxic drugs and inhibitors of insulin secretion on a serially transplantable rat insulinoma and cultured rat insulinoma cells. AB - The effects of cytotoxic drugs and inhibitors of insulin secretion were examined in vivo in rats with a radiation-induced transplantable insulinoma, and in vitro using cultured rat insulinoma cells and the derived RINm5F insulin-secreting cell line. Administration of diazoxide to insulinoma-bearing rats resulted in a transient decrease of plasma insulin with a temporary rise of glucose concentrations. Mannoheptulose and somatostatin failed to affect the marked hyperinsulinaemia and hypoglycaemia. Streptozotocin produced a rapid and sustained decrease of insulin concentrations in insulinoma-bearing rats, accompanied by a progressive elevation of plasma glucose. Administration of alloxan failed to affect circulating insulin or glucose concentrations. In vitro, streptozotocin and alloxan exerted approximately equipotent time-dependent and concentration-dependent cytotoxic effects on insulinoma cells and RINm5F cells as established by cell staining with trypan blue. The cytotoxic actions of both drugs were decreased by agents believed to scavenge free radicals or to act as inhibitors of poly(ADP-ribose) synthetase. The results suggest that the cytotoxic actions of streptozotocin and alloxan on rat insulinoma cells and RINm5F cells are mediated by the generation of hydroxyl free radicals and DNA strand breaks. The ineffectiveness of alloxan in insulinoma-bearing rats probably reflects the high rate of decomposition of the drug in vivo. PMID- 3032738 TI - Polymorphism and gene conversion in mouse alpha-globin haplotypes. AB - We have cloned and characterized three distinct alpha-globin haplotypes obtained from inbred strains of the mouse, Mus domesticus. We report here the complete nucleotide sequence of the six alpha-globin genes that the haplotypes contain. Our analysis of these genes and those from one other previously described haplotype indicates that recurrent gene conversion events have played a major role in their history. The pattern of nucleotide substitutions suggests that conversions have occurred both within and between haplotypes. Limited segments of coding and noncoding DNA have been involved in these gene conversion events. In two of the haplotypes, the nonallelic genes of each maintain DNA sequence identity over discrete intervals and encode the same alpha-globin polypeptide. On the other hand, the coding regions of some genes have accumulated replacement changes that result in distinct alpha-globins. In one instance, these changes appear to reflect positive selection of advantageous mutations. PMID- 3032739 TI - [Physical mapping of the immunity region of phage phi 80]. AB - Mapping of the sites of cleavage of five restriction endonucleases (BglI, BglII, EcoRV, KpnI and PvuII) in the immunity region of bacteriophage phi 80 DNA is described. The order of the 27 restriction sites was established. This helped to localize the phi 80 cI gene within the 640 bp fragment of the immunity region. Recombinant plasmids carrying phi 80 "kil" function were constructed. The function is suppressed by the cI-coded repressor. The deletion AB43 which is present in the phi 80 vir mutant is located within the phi 80 DNA fragment carrying the cI gene. PMID- 3032740 TI - [Plasmid vectors of "insertion inactivation" of the trimethoprim resistance marker]. AB - We demonstrate the possibility of using the T4 phage frd gene as an insertion inactivation marker within pBR322, in plasmids with changing copy number and expression of foreign genes under control. The structural part of the frd genes contains unique recognition sites for EcoRI and SalI endonucleases. Transformants with recombinant plasmids carrying the frd gene grow on media with up to 500 mkg/ml trimethoprim, whatever the gene dosage. PMID- 3032741 TI - [Effect of mutations of Escherichia coli K-12 chromosome on the integration of bacteriophage Mu introduced into cells by infection or conjugation]. AB - We present the detailed research on the previously described Escherichia coli K 12 Mud- mutants with impaired development of bacteriophage Mu. The ability of Mu phage DNA to penetrate into mutant cells on infection was shown. If introduced into the cells or combined with mud mutation by recombination, the prophage may be induced, which results in phage Mu lythic development and phage burst from mutant cells. In the course of conjugative transfer into the mutant cells, within a DNA fragment of the lysogenic donor chromosome, MupAp1 prophage is not inherited by recombinants. At the same time, Mu prophage deficient in genes A and B, whose products are required for transposition, is inherited by the mutant with the usual frequency. These data enable us to conclude that the mud mutations disturb the stage of conservative transposition which is connected with the insertion of the Mu prophage into the chromosome, after excision from the linear DNA introduced into the cells via infection or conjugation. PMID- 3032742 TI - A latitudinal cline in P-M gonadal dysgenesis potential in Australian Drosophila melanogaster populations. PMID- 3032743 TI - A test for the role of natural selection in the stabilization of transposable element copy number in a population of Drosophila melanogaster. PMID- 3032744 TI - Major spontaneous genomic rearrangements in Haemophilus influenzae S2 and HP1c1 bacteriophages. AB - Physical maps constructed by the localization of the cleavage site of several restriction endonucleases have shown that the genomes of the Haemophilus bacteriophages S2 and HP1c1 exist in variant forms which differ in the molecular organization of the genomes. At least three regions of different organization of the bacteriophage chromosomes have been identified. The different types of molecular organization can be detected both in the DNA isolated from the mature phage particles and after integration of the phage DNA into the bacterial chromosome. PMID- 3032745 TI - Identification of Paramecium mitochondrial proteins using antibodies raised against fused mitochondrial gene products. AB - Paramecium aurelia mitochondrial (mt) DNA fragments carrying the coding regions for two proteins, P1 (in the region adjacent to the origin of replication) and COII (subunit II of cytochrome oxidase), were used to study mt gene expression. The sequence for the portion of mtDNA containing P1 has already been described [Pritchard et al., Gene 44 (1986) 243-253]. The complete nucleotide sequence of the portion containing the COII gene is presented here. An 18.5-kDa protein was produced in maxicells when a fragment containing a major portion of the sequence coding for P1 was used. This fragment and a fragment carrying the COII gene were cloned into the expression vector pTRPLE', and antibodies were raised against the resulting fusion proteins in an Escherichia coli lysate. Western blots of Paramecium mt extracts identified two proteins, one 21 kDa (COII) and the other 23.5 kDa (P1). The size of the P1 protein is in agreement with the size of the open reading frame in that region of mitochondrial DNA. Based on extensive amino acid homology to the Paramecium gene and limited homology to COII genes from other organisms, the COII gene in another ciliate, Tetrahymena pyriformis, was identified just upstream of the small subunit rDNA in previously published sequences (Schnare et al., 1986). The size of the COII gene and the homology with the COII gene from Tetrahymena suggest that ATC, ATT, GTG and GTC could be used as translational initiators in Paramecium mitochondria. PMID- 3032747 TI - Terminal inverted repeats of prokaryotic transposable element IS186 which can generate duplications of variable length at an identical target sequence. AB - The insertion element IS186, which resides in the chromosome of Escherichia coli K-12, is 1338 bp long. Its termini represent 23-bp perfectly inverted repeats, but a variant carries a mismatch at position 23. IS186 transposes preferentially into G + C-rich sequences and generates target duplications of variable length, even at the same integration site. PMID- 3032746 TI - Human renin gene of renin-secreting tumor. AB - A large amount of renin mRNA was found to be expressed in the juxtaglomerular cell (JGC) tumor, as determined by Northern analysis. We have isolated the long 5'-flanking region of the human renin gene from the tumor, and characterized the promoter region with respect to nucleotide (nt) sequence and mRNA transcription start point. Of two sets of CAAT and TATA box at 29 bp upstream from the capping site is demonstrated to be a functional promoter by the primer extension. The 1:6 kb sequence, containing the 5'-flanking region, exon 1, and part of the first intron, obtained from the tumor was in complete agreement with that of the clone from fetal liver, which does not produce renin. This indicates that abnormal expression of the human renin gene in the JGC tumor involves no major alteration in the primary structure within 1.2 kb of the 5'-flanking region. Within 1.2 kb of the 5'-flanking region, there are several nt segments exhibiting homology with the glucocorticoid, estrogen, and progesterone receptor-binding sites and enhancers. These structures may be related to the tissue-specific expression of the renin gene. PMID- 3032748 TI - Complete nucleotide sequence of the penicillin acylase gene from Kluyvera citrophila. AB - The penicillin acylase (PAC) from Kluyvera citrophila ATCC21285 has been purified to homogeneity and found to be composed of two non-identical subunits of 23 and 62 kDa, in contrast with the previous findings [Shimizu et al., Agr. Biol. Chem. 39 (1975) 1655-1661]. The nucleotide (nt) sequence of the K. citrophila pac gene contained in the 3-kb PvuI-HindIII fragment of pKAP1 [Garcia and Buesa, J. Biotechnol. 3 (1986) 187-195] has been determined, showing that it encodes a protein of 844 amino acid (aa) residues. The aa analysis of the N-terminal and C terminal sequences of the purified subunits showed that they were derived from a common precursor protein of 93.5 kDa, from which a signal peptide of 26 aa, responsible for the periplasmic translocation of the protein, and an internal connecting polypeptide of 54 aa, have been removed in the maturation of the PAC. The comparison of the nt sequences of the pac genes from K. citrophila and Escherichia coli ATCC11105 [Schumacher et al., Nucl. Acids Res. 14 (1986) 5713 5727] revealed 80% homology, suggesting a common ancestral pac gene origin. The results reported here should allow investigation of the unusual mechanism of maturation of this prokaryotic protein, as well as manipulation, using DNA recombinant techniques, of the catalytic properties of this industrially important enzyme. PMID- 3032749 TI - Chromosome analysis by two-dimensional fingerprinting. AB - A two-dimensional fingerprinting technique has been developed that allows large, cell genome-size DNA's to be analyzed by restriction endonuclease cleavage and separation of DNA fragments by agarose gel electrophoresis. Equations have been derived to determine the genome size and number of cleavage sites from analysis of the distribution of fragment lengths. Genome sizes of Escherichia coli strain JM101 and two strains of the mycoplasma Acholeplasma laidlawii measured by this method are in agreement with published values. Other uses of two-dimensional fingerprinting for studies of prokaryotic and eukaryotic genome structure and organization are described. PMID- 3032751 TI - Treatment planning for particle radiation therapy. AB - Fast neutrons beams from the new medically dedicated cyclotrons in the US have depth dose characteristics comparable to photon beams from a 6-MV linear accelerator, at best. Treatment planning will have specific difficulties related to the relatively increased radiosensitivities of the brain, spinal cord, lens of the eye, and salivary gland. Therefore, exploitation of the potential biologic advantages compared to high energy photons will extract the price of increased difficulties in treatment planning. Dosimetric advantages of protons and helium ions compared to high energy photons are real and make possible the high-dose irradiation of cancers immediately adjacent to sensitive critical normal structures. Treatment planning and delivery with a precision of less than 2 mm is necessary. Such methods are already operational. Particle radiation therapy facilities are national resources, which can help the clinical radiation oncologist in the unique management of a few, specific problems. PMID- 3032750 TI - Efficient oligodeoxyribonucleotide-directed deletion mutagenesis using pEMBL vectors: removal of early region introns from polyoma virus mutants. AB - We have used oligodeoxyribonucleotide-directed deletion mutagenesis to remove early region introns from polyoma virus mutants. To this end we compared single priming, double priming, and gapped duplex approaches using either priming at 37 degrees C or at the critical temperature. The gapped duplex approach, coupled with priming at the critical temperature, resulted in up to 70% yield of the desired product. In conjunction with the use of the pEMBL vector system this method was simplified to yield specific deletions from cloned large DNA fragments with high efficiency. The resulting mutant plasmids could be used directly for biological assays without retransformation or recloning. RNA and protein analyses showed that removal of the large T- or middle T-antigen introns from polyoma early region mutants dl23 and dl8 was specific and resulted in DNA competent for the synthesis of only one T antigen. PMID- 3032752 TI - [Methodologic approach to testing the functional status of the peripheral nervous system of chickens in evaluating the late neurotoxic effects of organophosphorus compounds]. PMID- 3032753 TI - [Human Papillomavirus infection and cancer of the cervix uteri]. PMID- 3032754 TI - Gastric and duodenal polyps in familial adenomatous polyposis: a prospective study of the nature and prevalence of upper gastrointestinal polyps. AB - One hundred patients with familial adenomatous polyposis have prospectively undergone gastroduodenoscopy to identify and characterise polyps found. Forty six patients had polyps in the stomach or duodenum. Thirty five patients had adenomas (33 in duodenum, two in stomach) and 26 patients had fundic gland polyps. Some of these patients had polyps in the stomach and the duodenum. Adenomas in the duodenum were present in 33% of patients studied with Gardner's syndrome variant (p = 0.04). Adenomas were also more common in older patients. As adenomas may be a precursor of adenocarcinoma, routine surveillance of the stomach and duodenum with gastroduodenoscopy is recommended in patients affected with familial adenomatous polyposis. PMID- 3032755 TI - Malignant fibrous histiocytoma of the vulva. AB - Malignant fibrous histiocytoma is a sarcoma that usually occurs in the extremities of elderly patients. Primary sarcomas of the vulva are extreme rarities. The most common vulvar sarcoma is leiomyosarcoma. Only two case reports on malignant fibrous histiocytoma in vulva have previously been published. It is a malignant tumor capable of metastasizing by hematogenous and lymphatic routes. The therapy of choice is radical surgery. PMID- 3032756 TI - Human papilloma viruses and cervical intraepithelial neoplasia: the role of colposcopy. AB - To evaluate the reliability of colposcopy for distinguishing flat condyloma from cervical intraepithelial neoplasia (CIN), 211 patients with abnormal cytology, colposcopical evidence of an atypical transformation zone (ATZ), and a histological diagnosis of flat condyloma or CIN were studied. Colposcopic evidence of surface abnormalities, the presence of satellite lesions, and an irregular Lugol's uptake were tentatively considered to be features of condyloma. Histologically, koilocytotic lesions with a disorganized cytologically atypical basal/parabasal layer and with atypical mitotic figures (AMFs) were considered to be CIN cases, and designated as CIN with koilocytosis (CIN K). At least two colposcopic features of condyloma found in 98 of 99 flat condylomas, were also found in 89 of 112 CINs. When colposcopic features were matched with histology for every directed biopsy site, they correlated strongly with koilocytosis, regardless of the degree of atypia in the lesion. Moreover, these features often occurred at the periphery of poorly differentiated or undifferentiated, high grade CINs, in areas histologically indistinguishable from flat condyloma. Thus, colposcopic features are not of predictive value in distinguishing flat condyloma from CIN, do not show correspondence to the lesional degree of atypia and cannot be fully related to the biological characteristics of the cervices in which they are found. These findings confirm that colposcopy cannot be considered to be a diagnostic method. PMID- 3032757 TI - Effect of dihydroergocryptine and dihydroergocristine on cyclic AMP accumulation and prolactin release in vitro: evidence for a dopaminomimetic action. AB - Dihydroergocryptine and dihydroergocristine, two C-9, 10-hydrogenated ergot alkaloids, inhibited in a concentration-dependent manner prolactin release and cyclic AMP accumulation in cultured anterior pituitary cells. The inhibitory effect of dihydroergocryptine was more potent and started at lower concentrations than that of dihydroergocristine. Haloperidol and pimozide, two dopamine receptor antagonists, completely abolished the inhibitory activity of the ergot alkaloids. The involvement of the adenylate cyclase-cyclic AMP system in the inhibitory action of the two compounds was demonstrated by the antagonism by pertussis toxin of the reduction of both prolactin release and cyclic AMP accumulation produced by dihydroergocryptine and dihydroergocristine. PMID- 3032758 TI - Propranolol for prevention of recurrent gastrointestinal bleeding in patients with cirrhosis: a prospective study of factors associated with rebleeding. AB - In a previous randomized trial, we demonstrated that propranolol prevented recurrent gastrointestinal bleeding in patients with cirrhosis. We have undertaken the present study in a new group of patients to ascertain the factors associated with rebleeding. Among 232 patients with cirrhosis admitted for gastrointestinal bleeding, 127 were included. They received propranolol orally at a dose reducing the heart rate by 25%. The median follow-up period was 682 days. The following factors were studied: cause of cirrhosis; severity of cirrhosis; hepatocellular carcinoma recognized after inclusion; compliance; persistent decrease in heart rate; dose of propranolol; alcohol abstinence; previous history of hemorrhage; time interval from hemorrhage to onset of propranolol administration, and source of bleeding. The percentage of patients free of rebleeding was 71% at 1 year and 57% at 2 years. Only five factors were significantly and independently associated with rebleeding: occurrence of hepatocellular carcinoma; lack of compliance; lack of persistent decrease in heart rate; lack of abstinence, and previous history of bleeding. In conclusion, this study confirms the results of our previous trial and suggests that certain factors play a role in the mechanism of rebleeding in patients receiving propranolol. PMID- 3032759 TI - Increased prolyl hydroxylase activity and collagen synthesis in hepatocyte cultures exposed to superoxide. AB - Primary monolayer cultures of rat hepatocytes at confluence were exposed to an exogenously added source of superoxide, and its influence on collagen synthesis was examined. Superoxide was generated by the addition of dihydroxyfumarate to the culture medium. Exposure of hepatocytes to dihydroxyfumarate greatly stimulated the activity of prolyl hydroxylase and the synthesis of collagen. A significant increase in prolyl hydroxylase activity was observed with 5 micrograms per ml dihydroxyfumarate in 24 hr relative to that in the untreated cultures. Maximum stimulation of greater than 3-fold compared to the control value was elicited by 25 micrograms per ml dihydroxyfumarate. When scavengers of superoxide such as superoxide dismutase and Cu(Lys)2 were added in the medium, the increase in prolyl hydroxylase activity induced by dihydroxyfumarate was nearly abolished. Experiments with actinomycin D indicated that synthesis of new RNA was involved in the stimulation of prolyl hydroxylase activity. Analysis of collagen synthesis in cultures exposed to dihydroxyfumarate also showed a marked increase compared to that of the untreated cultures. The presence of superoxide dismutase in the medium significantly reduced the increase in collagen synthesis. Our results indicate that superoxide mediates the stimulation of collagen synthesis in hepatocytes. These findings may provide a possible explanation for excess collagen formation during induced liver fibrosis. PMID- 3032761 TI - Regulatory modulation in hepatology. PMID- 3032760 TI - Establishment and characterization of a human combined hepatocholangiocarcinoma cell line and its heterologous transplantation in nude mice. AB - A human cell line (KMCH-1) derived from a surgical specimen of combined hepatocellular and cholangiocarcinoma has been established. The original tumor consisted of both hepatocellular carcinoma of the trabecular type and cholangiocellular carcinoma. This cell line has been maintained for 26 months through 75 passages. KMCH-1 cells show characteristics of adenocarcinoma on light and electron microscopy. They proliferate in culture in a pave stone arrangement. The doubling time of these cells at the 24th passage was 39 hr. Chromosome analysis revealed a chromosome number ranging from 60 to 98, with a modal number of 74. KMCH-1 cells produced tumors several months after subcutaneous and intraperitoneal transplantations into athymic nude mice. Histologically, the subcutaneous tumors were poorly differentiated adrenocarcinoma, while intraperitoneal tumors were moderately to well-differentiated adenocarcinoma. Hepatocellular carcinoma components were not observed. Thus, KMCH-1 cells demonstrate the feature of cholangiocellular carcinoma in vitro and in vivo. This KMCH-1 cell line is the first established combined hepatocellular and cholangiocarcinoma cell line and may contribute to further investigation of combined hepatocellular and cholangiocarcinoma. PMID- 3032763 TI - The expression of 3-fucosylated-N-acetyl lactosamine carbohydrate determinants in renal tumours. AB - The distribution in renal tumours of 3-fucosyl-N-acetyl lactosamine has been studied by using the monoclonal antibodies AGF 4.36 and AGF 4.48 and immunoperoxidase methods on tissue sections. Seven of 19 nephroblastomas and 12 of 30 renal cell carcinomas contained the epitope. In nephroblastomas the epitope was found on the terminals of type B tubules in six cases and in one case on the type A or neoplastic tubules. In renal carcinoma the antigen was found on the surface of tumour cells. The results suggest that in kidneys bearing nephroblastomas ureteric bud elements may grow into the tumour from the adjacent kidney. PMID- 3032762 TI - Purine degradative enzymes in circulating malignant cells of patients with chronic B cell neoplasia. AB - Previous reports have shown that the purine degradative enzymes adenosine deaminase (ADA), purine nucleoside phosphorylase (PNP) and ecto-5'-nucleotidase (5'NT), play an important role in the normal development of lymphocytes and that investigations of these enzymes are of value in defining subsets of lymphoid malignancies of T-cell origin. Pharmacological inhibition of one of these enzymes has been found to be an effective treatment for a few lymphatic neoplasia. We have studied the activities of the above enzymes in the circulating malignant cells of 25 patients with B-chronic lymphatic leukemia (B-CLL), four patients with B prolymphocytic leukemia (PLL), seven patients with leukemic centrocytic lymphoma (CC), 18 patients with hairy cell leukemia (HCL) and 16 patients with immunocytoma (IC). For comparison, the blasts of nine patients with 'common' acute lymphatic leukemia (cALL) and normal T (n = 12) and B (n = 8) cells were simultaneously investigated. Despite morphologic similarity, the leukemic cells of the chronic B cell malignancies demonstrate different enzyme patterns. B-CLL is characterized by very low activities of all the enzymes ADA, PNP and 5'NT. In the cells of HCL the highest values of PNP are found. The leukemic cells of IC are characterized by low levels of ADA but moderate levels of PNP and high levels of 5'NT. Thus some of the entities of B malignancies show typical enzyme patterns which might be of importance in defining maturation stages of the disease. The differences in these enzyme patterns can also be made use of in therapy with enzyme inhibitors such as deoxycoformycin. PMID- 3032764 TI - [A study on nucleic acid hybridization of human nasopharyngeal carcinoma with EBV W fragment probe]. PMID- 3032765 TI - [A study of antineoplastic activity of DHAQ on human adenocarcinoma cells of lung hepatoma and gastric cancer cells in vitro]. PMID- 3032767 TI - Changing MFH to FHS. PMID- 3032766 TI - Thorotrast-associated hepatic leiomyosarcoma and cholangiocarcinoma in a single patient. AB - A 66-year-old man developed two distinct primary malignant hepatic neoplasms (leiomyosarcoma and cholangiocarcinoma) 50 years after Thorotrast administration. This is the first case report of Thorotrast-associated primary hepatic leiomyosarcoma. Ultrastructural and immunohistochemical studies are discussed. PMID- 3032768 TI - Chromosomal location by in situ hybridization of the human Sau3A family of DNA repeats. AB - The Sau3A family is a human, clustered, highly repetitive, GC-rich DNA family. In situ hybridization studies with a plasmid carrying a Sau3A monomer as a probe have shown that Sau3A sequences are preferentially concentrated in the heterochromatic regions of human acrocentric chromosomes (D and G groups, both in pericentromeric regions and in cytological satellites) and in pericentromeric heterochromatin of chromosome 1. The same chromosomal locations were observed by using as probes two recombinant phages which carry Sau3A-positive genomic sectors. The two sectors differ for the relative proportions of monomer and multiples of Sau3A repeats, which show different extents of homology to the cloned monomer, and for the presence, in one of the two, of a small amount of an unrelated repeat (alphoid DNA). The similarity of the results obtained with the three probes suggests that heterogeneous Sau3A repeats share the same chromosomal localizations and that the two analyzed genomic sectors may not contain significant amounts of repetitive DNAs other than the Sau3A family. A comparison between the chromosomal locations of Sau3A and EcoRI families of repeats has confirmed that each family is characterized by specific chromosomal locations and that single heterochromatic regions may contain both. PMID- 3032769 TI - Construction of a human ventricular cDNA library and characterization of a beta myosin heavy chain cDNA clone. AB - We have constructed and characterized for the first time a complementary DNA (cDNA) clone, pHMC3, which codes for a cardiac myosin heavy chain mRNA from human heart. This clone contains a 1.7 kb DNA segment and specifies 543 amino acids of the carboxyl portion of the myosin heavy chain. The DNA sequence and encoded amino acid sequence were compared to the hamster alpha (pVHC1) and beta (pVHC2/pVHC3) cardiac myosin heavy chain cDNA and amino acid sequences and the rat cardiac myosin heavy chain sequences as well. The myosin heavy chain mRNAs are highly conserved and this is reflected in our cDNA clone. The pHMC3 clone is 87.9% homologous to the hamster alpha cDNA and 92.2% homologous to the hamster beta cDNA clones. The 3' untranslated region of pHMC3 is 64.1% homologous to the hamster beta clone while the hamster alpha myosin heavy chain shows only 25% homology to pHMC3 and exhibits extensive diversity. Similar results were obtained when pHMC3 was compared to the rat cardiac myosin heavy chain cDNA sequences. The comparisons showed that pHMC3 is a beta cardiac myosin heavy chain cDNA clone. PMID- 3032770 TI - The 35 kd pulmonary surfactant-associated protein is encoded on chromosome 10. AB - The genomic components identified by each of two closely related cDNA clones for the major 35 kilodalton non-serum surfactant-associated proteins (PSP-A) were shown to derive from human chromosome 10 by Southern blot analysis of DNAs from human-rodent somatic cell hybrids. By in situ hybridization to human metaphase chromosomes, the cDNA probes were localized to the region 10q21-q24. PMID- 3032771 TI - Human ferritin H and L sequences lie on ten different chromosomes. AB - In humans, the H (heavy) and L (light) chains of the iron-storage protein ferritin, are derived from multigene families. We have examined the chromosomal distribution of these H and L sequences by Southern analysis of hybrid cell DNA and by chromosomal in situ hybridization. Our results show that human ferritin H genes and related sequences are found on at least seven different chromosomes while L genes and related sequences are on at least three different chromosomes. Further, we have mapped the chromosomal location of expressed genes for human H and L ferritin chains and have found an H sequence which may be a useful marker for idiopathic hemochromatosis. PMID- 3032772 TI - Signal transduction in human T lymphocytes. PMID- 3032774 TI - T-lymphocyte activation: the role of protein kinase C and the bifurcating inositol phospholipid signal transduction pathway. PMID- 3032773 TI - B-lymphocyte signal transduction in response to anti-immunoglobulin and bacterial lipopolysaccharide. PMID- 3032775 TI - Inhibition of mevalonate kinase by disulphide compounds. PMID- 3032776 TI - Fine needle aspiration cytology in diagnosis of hepatic malignancy. PMID- 3032777 TI - Murine cytomegalovirus. AB - BALB/c and C57BL/6 mice were infected with murine cytomegalovirus (MCMV). On day 4 or 12 of the infection, the animals were immunized with SRBC (T-dependent), TNP Ficoll (T-independent) and standard poliovirus. The adverse effect of the virus infection on humoral immune responses was limited to animals immunized on day 4; while anti-SRBC antibody formation was severely depressed in both mouse strains, reduced plaque forming cells to TNP-Ficoll were registered only in BALB/c mice. Antibodies to poliovirus were depressed in both strains, although to a lesser degree in C57Bl/6 than in BALB/c mice. Anti-SRBC B cell memory was found to be affected by MCMV infection. These results are interpreted to mean that T dependent and -independent antigens may be handled differently by the two mouse strains tested. PMID- 3032778 TI - The relationship between liver-specific lipoprotein and the hepatocyte plasma membrane. AB - Liver-specific lipoprotein (LSP) has been the subject of intense investigation as a candidate target antigen in chronic active hepatitis. Fundamental to the interest in LSP has been the belief that it is an antigen complex of hepatocyte plasma membrane origin. In this study the physical, biochemical and antigenic relationships between LSP and isolated hepatocyte plasma membrane (HPM) were investigated. Electron microscopic examination of LSP showed it to be devoid of plasmalemma sheets that were abundant in HPM. The plasma membrane marker enzyme 5'-nucleotidase was enriched 11-fold in HPM relative to liver homogenate, whereas the enzyme activity in LSP was 17% of that found in liver homogenate and only 1.5% of that found in HPM. The antigenic relationship between LSP and HPM was assessed using sera from rabbits immunized with either mouse LSP or mouse HPM. By filtration ELISA, antibody to LSP reacted poorly with entrapped HPM, relative to antibody to mouse HPM. Antisera to LSP and HPM were both effectively absorbed by the immunizing antigen, however antibody to LSP was not absorbed with HPM, and minimal cross-absorption of HPM antibody with LSP was found. By immunoblot of SDS PAGE separated LSP and HPM, it was shown that antigenic cross-reactivity between LSP and HPM at the polypeptide level was rare. By immunofluorescence, antibody to LSP failed to react with the surface of viable mouse hepatocytes, whereas antibody to HPM showed linear fluorescence. The data show that the two preparations, LSP and HPM, are dissimilar antigen complexes. HPM may be a more appropriate preparation for the study of autoimmune liver disease than LSP. PMID- 3032779 TI - Interleukin-1-like activity in capsular material from Haemophilus actinomycetemcomitans. AB - This paper describes the activity of a bacterial surface component (capsular material, CM) in biological assays for interleukin-1 (IL-1). CM from the periodontal pathogen Haemophilus actinomycetemcomitans was tested in the following in vitro assays: mouse thymocyte proliferation (LAF assay), stimulation of collagenase and prostaglandin (PG) E2 synthesis by articular chondrocytes, and stimulation of PGE2 synthesis by fibroblasts. In all these assays, CM gave a response similar to an IL-1 preparation. This ability to mimic IL-1 suggests an important role for CM in both cell-mediated immunity and connective tissue destruction in localized juvenile periodontitis (LJP). PMID- 3032780 TI - Release of leukotrienes during rapid expulsion of Trichinella spiralis from immune rats. AB - Rapid expulsion of the nematode Trichinella spiralis from immune rats is associated with an increase in volume of intestinal exudate and the presence of large numbers of tissue mucosal mast cells (MMC) and eosinophils. We have measured the concentrations of leukotrienes (LT) C4 (LTC4) and B4 (LTB4) in gut perfusates and mucosal homogenates at 30 min, 1, 3, 6 and 20 hr after challenge with larvae. Leukotrienes were identified by radioimmunoassay (RIA) combined with reverse-phase high-pressure liquid chromatography (RP-HPLC). There were significant elevations at 30 min and 1 hr in the concentrations of LTC4 in the perfusates from the gut of challenged immune rats compared to controls (infected unchallenged and uninfected naive rats). Similar increases in immunoreactive LTC4 and LTB4 were observed in mucosal homogenates from the gut of immune challenged animals. A second peak of LTB4 was also observed at 20 hr in both immune and naive challenged rats. There were also elevations in serum concentration of the MMC-associated specific serine protease, rat mast cell protease II (RMCPII). Since LTC4 causes smooth muscle contraction, increased vascular permeability and stimulation of mucus hypersecretion, and LTB4 recruits and activates inflammatory cells, leukotrienes may participate in the process of rapid expulsion of T. spiralis. PMID- 3032781 TI - Antibody-independent killing of gram-negative bacteria via the classical pathway of complement. AB - The experiments in this paper provided evidence that, besides lipopolysaccharides (LPS), porins of gram-negative bacteria bind to C1q and C1. From these experiments, we concluded that the association of LPS and porins (outer membrane proteins, OMP) may potentiate the C1q and C1 binding in the absence of specific antibodies. This antibody independent binding of C1 to LPS and porins is a prerequisite for the activation of the classical pathway of complement leading to the killing of serum-sensitive bacteria. PMID- 3032782 TI - C1 activation and dissociation in disease. AB - The composition of complexes containing C1 inactivator (C1 IA), C1r and C1s was investigated in normal serum after activation of C1 under various conditions. Analyses were performed with PAGE of eluates from Sepharose beads coated with F(ab')2 fragments of anti C1s followed by immunoblotting with anti C1 IA, anti C1s or anti C1r. Eluates obtained from serum treated with aggregated IgG (AGG) contained C1 IA in complex with C1r and C1s with both subcomponents in activated form. Eluates from serum incubated at 37 degrees C for 1, 2 or 3 days without activators showed C1 IA complexed with activated C1r and with C1s in proenzyme state associated to the complex. On analysis of serum, treated as mentioned above, by a variant of the electroimmunoassay using an intermediate gel containing anti-C1 IA and with anti-C1s in the anodal gel the two types of C1r- C1s--C1 IA complexes could be distinguished. Investigation of fresh sera and synovial fluids from patients with rheumatoid arthritis in this assay showed complexes containing C1 IA and C1r-C1s in activated form in the synovial fluids, while C1 IA-activated C1r-proenzyme C1s complexes were found in the corresponding sera. PMID- 3032783 TI - The endogenous retrovirus Mtv-8 on mouse chromosome 6 maps near several kappa light chain markers. AB - Endogenous retroviruses are known to affect expression of cellular genes in the vicinity of their integration sites. The endogenous mouse mammary tumor provirus (Mtv-8) previously has been reported to reside on mouse chromosome 6 near the immunoglobulin kappa chain locus. Using pairs of mouse strains on the BALB/c (Mtv 8 positive) and C58 (Mtv-8 negative) backgrounds which are congenic for chromosome 6 genetic markers, we have confirmed the chromosome assignment of this provirus. Moreover, we have analyzed the N1 progeny of a (B6 X C58) X C58 backcross to determine the segregation of the Mtv-8 provirus with respect to polymorphisms in the Igk-VSer and Igk-J loci. The results with congenic and backcross mice together with results of others suggest that Mtv-8 is located approximately 0.52 cM from several closely linked kappa markers on chromosome 6. PMID- 3032784 TI - Isolation, expression, and the primary structure of HLA-Cw1 and HLA-Cw2 genes: evolutionary aspects. AB - The HLA-Cw1 and -Cw2 genes were identified in a genomic library and their products characterized by biochemical methods. The HLA-Cw1 and -Cw2 genes, upon transfection in mouse L cells, give rise to class I antigen heavy chains that associate with neither mouse nor human beta-2 microglobulin. They are indistinguishable in isoelectric point from polypeptides identified as HLA-Cw1 and -Cw2 in human cells. The nucleotide sequence of HLA-Cw1 and -Cw2 and their comparison with HLA-Cw3, the only other known HLA-C sequence, reveal a characteristic pattern of locus-specific amino acids. A comparison of 13 different human class I primary structures leads us to speculate that the most variable region in HLA class I antigens, positions 61-83, could assume an alpha helical structure of critical importance for class I antigen function. The locus specificity and the higher degree of intralocus conservation in the COOH-terminal region, especially in the transmembrane and cytoplasmic domains, must reflect evolutionary ancestry rather than positive selection. In view of the pattern and types of substitutions observed for HLA-C locus products, their function as immune response gene products is questioned. PMID- 3032785 TI - Characterization of the human homolog of the rat MRC OX-2 membrane glycoprotein. AB - The MRC OX-2 antigen is a membrane glycoprotein present on rat thymocytes, neurons, follicular dendritic cells, endothelium, and some smooth muscle. The sequence of 248 amino acids has similarities to Ig domains organized with one V like domain, one C-like domain, and transmembrane and cytoplasmic regions. Thus it resembles a T-cell receptor chain but shows no sequence divergence. We report the characterization of the human gene for this molecule. Its exon organization is similar to that found for immunoglobulins although the region with similarities to Ig J regions is found within the same exon as the V-like domain. Human MRC OX-2 is expressed at the mRNA level in brain and B-cell lines but not detected in liver or T-cell lines. It does not obviously correspond to any previously defined leukocyte antigen. The sequence homology for the human and rat MRC OX-2 molecules is higher for the Ig-related region (75%) than for many other Ig-related molecules and very high in the transmembrane region (96%), implying a functional role other than simply its anchoring into the membrane. PMID- 3032787 TI - Interface mechanics and histology of titanium and hydroxylapatite-coated titanium for dental implant applications. PMID- 3032786 TI - Characterization of an HLA-DR beta pseudogene in the DRw52 supertypic group. AB - The nature of the DR beta II pseudogene in a haplotype of the DRw52 supertypic group was investigated by nucleotide sequence analysis. It revealed several deleterious mutations in the signal sequence and second domain regions in addition to the complete absence of the first domain and adjacent sequences. No expression of DR beta II pseudogene mRNA can be detected. The same DR beta II pseudogene is probably present in other members of the DRw52 supertypic group. The pattern of mutations in this DR beta II pseudogene is different from that observed in the DR beta pseudogene of the DRw53 supertypic group, indicating a distinct evolutionary pathway for these two groups of DR haplotypes. PMID- 3032788 TI - Red blood cell Na+,K+-ATPase in men with newly diagnosed or previously treated essential hypertension. AB - Alterations of cellular function of Na+,K+-adenosine triphosphatase (ATPase; Na+ K+ pump) have been implicated in the pathophysiology of essential hypertension. Therefore, this aspect of red blood cell (RBC) Na metabolism was studied in black men with newly diagnosed, untreated essential hypertension (NEH) and a normotensive control group. RBC Na content, Na+-K+ pump number (ouabain binding sites), and pump activity were measured. No statistically significant differences were found between the two groups for any of these three parameters. However, a group of previously treated essential hypertensive subjects (PEH) who had been withdrawn from therapy in the preceding 6 weeks were also studied. This group differed significantly from the NEH subjects in regard to all RBC Na+-K+ pump parameters. Their RBC Na content (10.27 +/- 3.23 vs 7.77 +/- 2.52 mmol Na/LRBC; p = 0.006) was higher, and their Na+-K+ pump activity (166 +/- 50 vs 221 +/- 87 nmol inorganic phosphate/mg membrane protein/hr; p = 0.03) and Na+-K+ pump number (213 +/- 40 vs 284 +/- 85 binding sites/RBC; p = 0.001) were lower compared with those in NEH subjects. Although the PEH subjects were older and somewhat less hypertensive than their NEH counterparts, these factors were not found to influence the Na+-K+ pump parameters. These results indicate that chronic diuretic therapy of patients with essential hypertension is associated with a reduced number of RBC Na+-K+ pumps. Since RBCs are not considered target cells for diuretics, the effects of these drugs on RBC electrolyte metabolism may occur at the time of erythropoiesis by the production of RBCs with fewer Na+-K+ pumps.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3032789 TI - Preserved cardiac beta-adrenergic sensitivity in early renovascular hypertension. AB - To determine the mechanism of blunted sympathetic reflex responses in early renovascular hypertension, we measured inotropic and chronotropic responses of the heart to beta-adrenergic stimulation in vivo and myocardial beta-adrenergic receptor number and adenylate cyclase activity in 10 dogs during an early stage of one-kidney renal hypertension. Mean aortic pressure was higher in the hypertensive dogs (152 +/- 4 mm Hg) than in eight sham-operated dogs (122 +/- 1 mm Hg; p less than 0.001), but heart rate, cardiac output, and left atrial pressure did not differ between the two groups. Blood pressure reduction with a direct-acting vasodilator, pinacidil, resulted in marked increases in heart rate (+97 +/- 12 beats/min) and rate of change of left ventricular pressure (dP/dt; +1447 +/- 367 mm Hg/sec) in normotensive dogs but only blunted heart rate (+54 +/ 12 beats/min) and minimal left ventricular dP/dt (+376 +/- 264 mm Hg/sec) responses in hypertensive dogs. In contrast, intravenously administered isoproterenol produced similar increases in heart rate and left ventricular dP/dt in the two groups. These two groups also did not differ in either left ventricular beta-adrenergic receptor number and affinity or basal, isoproterenol stimulated, and fluoride-stimulated adenylate cyclase activity. Thus, despite blunted reflex responses to blood pressure reduction, hypertensive dogs showed neither reduction in chronotropic and inotropic responses to direct beta adrenergic stimulation nor beta-adrenergic desensitization of the myocardium, as assessed by beta-adrenergic receptor number and adenylate cyclase activity. Blunted reflex responses in this model of early hypertension must be due to factors operating at some locus other than the beta-adrenergic receptor-adenylate cyclase complex. PMID- 3032790 TI - Cytochemically assayable Na+,K+-ATPase inhibition by Milan hypertensive rat plasma. AB - The ability of plasma from 3- and 9-week-old Milan hypertensive rats and their normotensive controls to inhibit Na+,K+-adenosine triphosphatase (ATPase) was studied using cytochemical bioassay techniques in fresh tissue. With a validated cytochemical bioassay that measures the capacity of biological samples to stimulate glucose-6-phosphate dehydrogenase activity in guinea pig proximal tubules as an indication of their capacity to inhibit Na+,K+-ATPase, the mean glucose-6-phosphate dehydrogenase-stimulating ability of the plasma of the 9-week old Milan hypertensive rats and their normotensive controls was 586.0 +/- 88 and 23.4 +/- 8.3 U/ml (n = 7; p less than 0.001), while that of the 3-week-old Milan hypertensive rats (before the main rise in arterial pressure) and their normotensive controls was 99.9 +/- 27.4 and 7.8 +/- 1.8 U/ml (n = 7; p less than 0.001). With the use of a semiquantitative cytochemical assay that measures Na+,K+-ATPase activity directly, plasma from the adult hypertensive rats had a much greater capacity to inhibit Na+,K+-ATPase than the plasma of the control rats. The significantly raised levels found in the young hypertensive rats before the main rise in arterial pressure are consistent with the hypothesis that the rise in the ability of plasma to inhibit Na+,K+-ATPase is due to an inherited renal difficulty in excreting sodium. PMID- 3032791 TI - Decreased density of vascular receptors for atrial natriuretic peptide in DOCA salt hypertensive rats. AB - We have previously found that vascular receptors for atrial natriuretic peptide (ANP) in the rat are down-regulated by volume expansion. For this reason vascular ANP receptor density and affinity were examined in a model of volume-expanded hypertension, the deoxycorticosterone acetate (DOCA)-salt hypertensive rat. The density of mesenteric vascular ANP binding sites was decreased in DOCA-salt hypertensive rats from a control value in uninephrectomized rats of 203 +/- 25 fmol/mg protein to 60 +/- 13 fmol/mg protein (p less than 0.01). The sensitivity of norepinephrine-precontracted aorta to ANP was significantly reduced in DOCA salt hypertensive rats (p less than 0.001). DOCA-salt hypertensive rats infused intravenously for 4 days with ANP, 100 to 300 ng/hr, did not experience a lowering of blood pressure, in contrast to the significant reduction in blood pressure seen in two-kidney, one clip Goldblatt hypertensive rats similarly infused. In the latter there was no natriuretic response to ANP, while in the DOCA-salt hypertensive rats natriuresis occurred without lowering of blood pressure. In the DOCA-salt hypertensive rats plasma ANP concentration was increased to 68 +/- 8 fmol/ml from 10 +/- 1 fmol/ml in uninephrectomized rats. In conclusion, raised ANP concentration in plasma of volume-expanded hypertensive rats (DOCA-salt hypertension) may result in decreased density of ANP vascular receptors. These results suggest that a decrement in the number of ANP receptors may be a cause of decreased sensitivity of vascular responses to ANP in vitro and resistance to the blood pressure-lowering action of ANP in vivo. PMID- 3032792 TI - In vitro inhibition of group B streptococcus-induced polymorphonuclear leukocyte aggregation. AB - Evidence suggests that part of the pathophysiologic response seen in group B streptococcal (GBS) sepsis may be due to polymorphonuclear leukocyte (PMN) activation. Indomethacin (INDO), which inhibits eicosanoid metabolism, attenuates the pathophysiologic response stimulated by GBS, possibly due to inhibition of PMN aggregation. We examined the capability of two eicosanoid metabolism inhibitors, INDO and nordihydroguaiaretic acid (NDGA), to inhibit PMN aggregation induced by heat-inactivated opsonized GBS and GBS-activated plasma. Opsonized GBS induced PMN aggregation was inhibited by INDO (50-500 microM) and NDGA (1-100 microM). Over similar concentration ranges, INDO and NDGA had no significant effect on PMN aggregation induced by GBS-activated plasma. PMNs in plasma aggregate in response to unopsonized GBS. The stimuli for aggregation are opsonized GBS and GBS-activated plasma. INDO (50-500 microM) was unable to inhibit aggregation under this condition. Over the same concentration range in which INDO inhibited opsonized GBS-induced PMN aggregation, INDO was unable to inhibit opsonized GBS-induced superoxide production in PMNs. NDGA was examined but was found to interfere with the assay. The above evidence suggests PMN aggregation via eicosanoid metabolism may play a role in GBS-induced sepsis, which may be attenuated by agents such as INDO and NDGA. PMID- 3032793 TI - 111Indium-tropolone labeled human PMNs: a rapid method of preparation and evaluation of labeling parameters. AB - Pure polymorphonuclear leukocytes (PMNs) have been isolated from a small amount of human blood by a single-step density gradient centrifugation method using a commercially available Ficoll-Hypaque mixture of density 1.114. The cells were labelled with [111In]tropolone in both buffer and plasma. Cell viability, ability to generate superoxide anion, and chemotaxis were found to be unaltered both before and after labeling. The optimum tropolone concentration for labeling was found to be 1 X 10(-4) M. Labeling efficiency was higher at 37 degrees C than at room temperature. Compared to [111In]oxine, tropolone preparation both in buffer and plasma resulted in consistently higher yields. Preliminary experiments of in vivo cell viability of the labeled PMNs were carried out in rabbits. The ability of the cells to localize in experimentally produced inflammatory lesions was found to be intact. The method of cell separation and labeling described has been found to be simple and rapid and could easily be incorporated in routine nuclear medicine laboratory practice. PMID- 3032795 TI - Alterations in neutrophil superoxide production following piroxicam therapy in patients with rheumatoid arthritis. AB - Twenty patients with classical rheumatoid arthritis were enrolled in a study to determine the effects of piroxicam therapy on neutrophil function as defined by chemotaxis and superoxide anion (O2-) production. T-lymphocyte chemotaxis was also evaluated in these patients. Leukocytes were obtained from these subjects initially, after two weeks of placebo treatment, and subsequently after four and 10 weeks of piroxicam therapy (20 mg, once daily). Responses were compared to simultaneously tested normal controls and to the patient's own cells obtained at the different time points. Studies showed that four and 10 weeks of piroxicam therapy resulted in significantly suppressed neutrophil O2- production in response to phorbol myristate acetate (PMA) and formyl methionyl leucyl phenylalanine (FMLP). O2- production in response to opsonized zymosan was not significantly affected after four weeks of therapy, but was significantly reduced after 10 weeks of therapy when compared to the patient's own cell response after two weeks of placebo treatment. Unlike O2- production, PMN random migration and chemotaxis in response to C5a or FMLP did not differ significantly from normal or untreated patient controls. Analysis of T-lymphocyte migration showed that T-cell random migration or migration to the chemokinetic agent, casein, was not significantly altered by piroxicam therapy. However, when T lymphocytes were tested for chemotaxis in response to lymphocyte-derived chemotactic factor for T cells (LCF), T cell migration was significantly suppressed after 10 weeks of therapy. Furthermore, when T cells from these subjects were cultured for 24 h, random migration was significantly reduced after four and 10 weeks of piroxicam therapy when compared to the patient prior to therapy, and migration in response to LCF was suppressed after four weeks of therapy when compared to normal controls. These data indicate that in patients with rheumatoid arthritis, treatment with piroxicam will significantly suppress PMN O2- production and may also alter the locomotor capacity of T lymphocytes. These actions may contribute to the antiinflammatory effects of piroxicam. PMID- 3032794 TI - Stimulated release of neutral proteinases elastase and cathepsin G from inflammatory rat polymorphonuclear leukocytes. AB - Rat leukocytes from inflammatory peritoneal exudates respond in vitro to a variety of chemotactic and phagocytic stimuli by releasing both elastase and cathepsin G neutral proteinase enzyme activities. PAF, FMLP, and PMA stimulated a rapid, cytochalasin B-dependent, dose-related release of both enzymes; however, leukotriene B4 was inactive. It was not possible to measure the activity of zymosan-activated serum on these cells as rat serum contains high levels of proteinase inhibitors. The calcium ionophore A23187 stimulated a dose-related, time-dependent, cytochalasin B-independent enzyme release. Concanavalin A stimulated a weak, nondose-related release of enzyme activity. Zymosan and serum coated zymosan stimulated enzyme secretion which was markedly inhibited by the presence of cytochalasin B. These data indicate that release of azurophillic granule neutral proteinases from rat inflammatory leukocytes can be detected and quantitated in vitro. This model could provide a test system for monitoring the pattern and specificity of enzyme release from azurophil granules. The ability of a variety of stimuli to induce proteolytic enzyme release from inflammatory neutrophils may be of considerable relevance to chronic inflammatory diseases. PMID- 3032796 TI - Role of high-avidity binding of human neutrophil myeloperoxidase in the killing of Actinobacillus actinomycetemcomitans. AB - The binding of the neutrophil enzyme myeloperoxidase (MPO) to microbial surfaces is believed to be the first step in its microbicidal activity. The MPO-H2O2-Cl- system is responsible for most oxidative killing of Actinobacillus actinomycetemcomitans by human neutrophils. There appear to be three forms of MPO (MPO I, II, and III), all of which can kill this organism in the presence of H2O2 and chloride. In this study, we characterized the binding of native human neutrophil MPO to A. actinomycetemcomitans by an elution procedure dependent on the cationic detergent cetyltrimethylammonium bromide. Binding of native MPO was rapid and reached apparent equilibrium within 1 min. A proportion of binding under equilibrium conditions was saturable and highly avid, with a capacity of 4,500 sites per cell and a dissociation constant of 7.9 X 10(-10) M. At equal protein concentrations, more MPO III bound than MPO II, and more MPO II bound than MPO I. The high-avidity interaction was inhibitable with yeast mannan and with the serotype-defining mannan of A. actinomycetemcomitans. Binding was also partially reversible with yeast mannan. MPO bound to the high-avidity sites did not oxidize guaiacol but oxidized chloride, as detected by the chlorination of taurine. MPO bound to the high-avidity sites was incapable of killing A. actinomycetemcomitans alone in the presence of H2O2 and Cl-, but potentiated killing when sufficient additional MPO was provided. The killing of A. actinomycetemcomitans by the MPO-H2O2-Cl- system was inhibited by yeast mannan and a serotype-defining mannan of A. actinomycetemcomitans. We conclude that high avidity binding of MPO to the surface of A. actinomycetemcomitans is a mannan specific interaction and that MPO bound to the high-avidity sites is essential but not alone sufficient to kill A. actinomycetemcomitans. PMID- 3032798 TI - Detection of nucleoside triphosphate hydrolase as a circulating antigen in sera of mice infected with Toxoplasma gondii. AB - The nucleoside triphosphate hydrolase, which is present in the tachyzoite form of Toxoplasma gondii, was detected as a circulating antigen in sera of mice infected with a virulent (RH) or an avirulent (Beverly) strain of T. gondii. The enzyme was detected with a monoclonal antibody incorporated into an enzyme-linked immunosorbent assay. The lower limit of sensitivity of the assay was about 0.3 ng/ml, and standard assays provided a linear plot of nucleoside triphosphate hydrolase concentration over a range of 0.3 to 12 ng/ml. In mice inoculated intraperitoneally with tachyzoites of the RH strain, nucleoside triphosphate hydrolase emerged in the serum 1 day after injection, and then the concentration increased and reached a value of 30 micrograms/ml on day 5. In mice inoculated intraperitoneally with cysts of the Beverly strain, nucleoside triphosphate hydrolase was detected at day 3 after injection, and a peak concentration of 89 ng/ml was seen on day 10. The concentration of enzyme decreased thereafter, and the enzyme disappeared from the circulation on day 56. PMID- 3032797 TI - Role of myeloperoxidase in the killing of Naegleria fowleri by lymphokine-altered human neutrophils. AB - Previously we have shown that human neutrophils treated with conditioned medium from phytohemagglutinin-stimulated mononuclear leukocytes (sCM) in the presence of antisera have amoebicidal properties for Naegleria fowleri, a pathogenic free living amoeba. The data now presented show that neutrophils which lack myeloperoxidase (MPO) but have a normal oxygen-dependent respiratory burst could not be altered by sCM to express the amoebicidal activity. Catalase inhibited this amoebicidal activity of sCM-treated neutrophils. Various components and products of the neutrophils were examined for effects on naegleriae. A granule extract was found to have no effect at concentrations up to 100-fold that which killed Salmonella minnesota R595. Hydrogen peroxide appeared to have little effect even at 100 microM. However, in the presence of MPO, H2O2 was amoebicidal at 2.5 microM. The generation of amoebicidal activity required the presence of chloride ions. Azide inhibited the effects of the MPO-H2O2-Cl- system. Arginine, a scavenger of hypochlorite, significantly depressed the ability of sCM-treated neutrophils to kill amoebae and also prevented the amoebicidal properties of the MPO-H2O2-halide system. These results suggest that the MPO-H2O2-halide system is important in the killing of naegleriae by sCM-treated neutrophils and that hypochlorite may be the amoebicidal agent. PMID- 3032799 TI - Morphogenesis and pathogenicity of Histoplasma capsulatum. AB - The sulfhydryl blocking agent p-chloromercuriphenylsulfonic acid (PCMS) irreversibly inhibited the mycelium-to-yeast transitions of two virulent strains of Histoplasma capsulatum, G184A and G222B, when the temperature of incubation was raised to 37 degrees C, and the block persisted even after the cultures were washed free of PCMS. Instead of transforming to yeast cells, PCMS-treated mycelia continued to grow as mycelia at the elevated temperatures. A less virulent strain (Downs) was more temperature sensitive, but it showed a similar irreversible effect at 34 degrees C. Therefore, the mycelium-to-yeast transition of H. capsulatum is not required for the adaptation of mycelia to elevated temperatures but probably results from the temperature-dependent activation of yeast-specific genes. The transition to yeast is inferred to be obligate for pathogenicity in mice because PCMS-treated mycelia failed to cause infection, and no fungi were seen in tissues after PCMS-treated mycelia were injected into mice. PMID- 3032800 TI - Opsonization of Staphylococcus aureus protects endothelial cells from damage by phagocytosing polymorphonuclear leukocytes. AB - When phagocytosis of Staphylococcus aureus by human polymorphonuclear leukocytes (PMN) takes place on the surface of cultured human endothelial cells, the endothelial monolayers are damaged by lysosomal enzymes that are released by the PMN. Because PMN can phagocytose opsonized as well as unopsonized staphylococci on an endothelial surface, we studied the role of bacterial opsonization in the damage caused to the endothelium. Phagocytosis of unopsonized S. aureus was accompanied by greater damage (expressed as the percentage of the endothelial cells detached from the culture plates) of the monolayers than was phagocytosis of opsonized S. aureus: 52 +/- 10% and 24 +/- 7%, respectively, after 30 min of phagocytosis and 73 +/- 5% and 50 +/- 6%, respectively, after 60 min of phagocytosis. When correlated to the amount of phagocytosis, this difference was even greater (uptake was 35 +/- 4% for unopsonized S. aureus and 56 +/- 5% for opsonized S. aureus after 30 min and 42 +/- 3% and 60 +/- 5%, respectively, after 60 min). Total release of lysozyme and myeloperoxidase and generation of superoxide anion were the same during phagocytosis of opsonized or unopsonized staphylococci. Adherence of PMN to the endothelial cells was greater during phagocytosis of unopsonized S. aureus: 42 +/- 4% verus 27 +/- 3% during phagocytosis of opsonized staphylococci. Possibly, increased adherence of the PMN resulted in a locally higher concentration of enzymes which induced more damage. We conclude that opsonization of bacteria not only improves bacterial uptake, but also protects bystander cells from damage by the phagocytosing PMN. PMID- 3032801 TI - Effect of crevicular fluid and lysosomal enzymes on the adherence of streptococci and bacteroides to hydroxyapatite. AB - Samples of hydroxyapatite (HA) beads smaller than 1 mg were coated with 10 microliter of either saliva, serum, or human crevicular fluid before being added to a suspension of Streptococcus sanguis or Bacteroides gingivalis. In some assays, preparations of a granular fraction, elastase, or cathepsin G from human leukocytes were used to coat HA or to treat saliva-coated HA (SHA) before mixing with bacteria. The number of cells adhering to the beads was then counted under a scanning electron microscope by a standardized procedure. More cells were found to adhere to SHA in this assay than in the conventional large-scale assay. Human crevicular fluid, even when diluted up to three times, completely inhibited the adherence of S. sanguis to HA. A 100% inhibition of S. sanguis adherence was also observed when HA was coated with the granular fraction of leukocytes, and a 65% inhibition observed when SHA was treated with the enzyme preparation. When used to coat HA, elastase and cathepsin G reduced the adherence of S. sanguis by 30 and 50%, respectively. The binding of S. sanguis to elastase- or cathepsin G treated SHA was also reduced. B. gingivalis 33277 was found to adhere in high numbers to SHA. Coating HA with crevicular fluid or with the lysosomal enzyme preparation had a limited negative effect. We postulate that crevicular fluid prevents the adherence of S. sanguis by virtue of either its enzyme content or its albumin content or both. PMID- 3032803 TI - HTLV-III/LAV and the origin and pathogenesis of AIDS. PMID- 3032802 TI - Staphylococcal enterotoxin B as a nonimmunological mast cell stimulus in primates: the role of endogenous cysteinyl leukotrienes. AB - The immediate-type skin reaction and the emetic response in unsensitized monkeys on challenge with staphylococcal enterotoxin B (SEB) were studied to define the role of cysteinyl leukotrienes (LTs) in the action of the toxin. LY 171883, a selective LTD4/LTE4 receptor inhibitor, antagonized SEB-induced skin reactions and emetic responses completely. Inhibition of prostanoid formation by indomethacin, however, and pretreatment with BW 755C, a dual lipoxygenase and cyclooxygenase inhibitor, did not influence these reactions. The generation of endogenous cysteinyl LTs upon intragastric SEB administration was established in vivo. There was a tenfold increase in LTE4, the major biliary cysteinyl LT, and a novel cysteinyl LT metabolite in urine occurred, indicating strongly enhanced LT generation on SEB challenge. These results provide the first evidence that cysteinyl LTs may be important mediators in the pathophysiology of SEB-induced effects, as a model for pseudo-allergic reactions. PMID- 3032804 TI - The development of a new agent for the treatment of inflammatory/allergic conditions. AB - The search for new drugs for the treatment of reversible obstructive airways disease has been a major pursuit of the pharmaceutical industry for almost two decades. Little progress has been achieved. The acceptance that asthma is a multi facetted disease state has led to the development of a new complex in vivo screen in the macaque monkey. This model is mediated through mast cells which stain positively with alcian blue/safranin. A new therapeutic agent, nedocromil sodium, was significantly more active than sodium cromoglycate in this primate model of airway disease. Extensive clinical trials have shown that this new agent is effective in the treatment of the reversible component of obstructive airways disease. PMID- 3032805 TI - Betamethasone modulates the biological function of human polymorphonuclear leukocytes. AB - We have examined the influence of betamethasone (BT) on cyclic AMP (cAMP) metabolism and lysosomal enzyme release from highly purified (approximately equal to 99%) human polymorphonuclear leukocytes (PMNs). Preincubation (1-24 h) of human PMNs with BT (10(-9)-10(-5) M) had no effect on either cAMP content or on beta-glucuronidase release induced by formyl-containing tripeptide (f-met peptide). Preincubation (16-24 h) of PMNs with BT (10(-8)-10(-7) M) dose dependently potentiated the cAMP accumulation caused by beta-agonists (isoproterenol), adenosine A2/Ra agonist (NECA), prostaglandin E1 (PGE1) and histamine in PMNs. Similarly, BT potentiated the inhibition of f-met peptide induced beta-glucuronidase release from human PMNs caused by PGE1 (10(-6) M), histamine (2 X 10(-5) M), NECA (10(-4) M) and isoproterenol (10(-6) M). PMID- 3032808 TI - Effect on leukocyte locomotion and superoxide production by uremic toxins and polyamines. AB - We have isolated peak II, which is the peak of the middle molecules containing polyamines, from dialysate of uremic patients in recirculating dialysis. We investigated the effect of total dialysate, chromatographic peaks, I, II, III, IV and commercial polyamines on polymorphonuclear leukocyte chemotaxis and superoxide production (i.e. phagocytic activity) in healthy and uremic patients. Polymorphonuclear chemotaxis was inhibited by total dialysate and peak II but not by polyamines; polyamines, total dialysate and peak II had no activity on polymorphonuclear phagocytosis. PMID- 3032807 TI - Buffers in peritoneal dialysis. PMID- 3032806 TI - Effects of some prostaglandins and leukotrienes on lymphocytes, monocytes and their activity in vitro. AB - The effect of some prostaglandins (PGs) and leukotrienes (LTs) on lymphocytes and monocytes was tested in vitro. Overnight incubation with PGD2 reduces the expression of Leu-2 antigen (suppressor/cytotoxic phenotype) and of Fc receptors for IgG. PGA2, PGD2 and to a lesser extent PGE2 inhibit PHA reactivity of lymphocytes. LTB4 does not have any inhibitory activity. Preincubation of lymphocytes with PGD2 and PGE2 decreases their NK activity. LTB4, but not the PGs tested, stimulates monocyte metabolism as assessed by chemiluminescence. PMID- 3032809 TI - Human papillomavirus type 17 transcripts expressed in skin carcinoma tissue of a patient with epidermodysplasia verruciformis. AB - Certain types of human papillomavirus (HPV) are involved in skin carcinogenesis in epidermodysplasia verruciformis. However, no gene or gene product of HPV associated with skin carcinogenesis has yet been identified. Here, we report HPV 17 transcripts expressed in skin carcinoma tissue of an epidermodysplasia verruciformis patient infected with HPV-17. Further, we show that one of these transcripts was localized to a portion of the genome which contains the 3' open reading frames of the early region (E2, E3, E4 and E5). The analogous region in bovine papillomavirus type I has been shown to contain a transforming gene (Nakabayashi et al., 1983; Sarver et al., 1984; Yang et al., 1985a). PMID- 3032810 TI - Ultrastructural evidence of collagenolytic activity in ductal infiltrating carcinoma of the human breast. AB - The stroma of ductal infiltrating carcinoma of the human breast shows characteristic and localized areas of collagen rarefaction and fragmentation. This finding has been correlated with a peculiar type of fibrillar damage, observed in a small percentage of collagen fibrils isolated in the native state from the tumour stroma. The same pattern of lesion has been reproduced in vitro by human collagenase digestion on reconstituted fibrils. No effect has been detected by other nonspecific proteases in the same system. PMID- 3032811 TI - A new animal model for human colon cancer metastasis. AB - An animal model for human colon cancer metastasis is described in which spontaneously metastasizing colonic tumors are formed after injection of human colon cancer cells into the cecal wall of young athymic nude mice. Lymphatic and vascular invasion were demonstrated histologically after injection of both well- and poorly-differentiated cell lines, and metastases were found in a pattern similar to that of naturally occurring human colonic cancer. In contrast, little or no visceral organ involvement could be demonstrated after s.c. injection. Cells with increased liver-metastasizing potential were obtained by serial selection in this system. These cells had an enhanced ability to penetrate a reconstituted basement membrane in the presence of partially purified liver extract when compared to lung or brain extracts in a modified Boyden chamber assay. These results demonstrate the ability of human epithelial tumor cells to metastasize reproducibly in an animal model system, which may be useful for studying many aspects of the pathogenesis of cancer metastasis. In addition, it is suggested that local invasion by colon cancer cells may be influenced in part by tissue-specific factors. PMID- 3032812 TI - Practical guidelines for the use of converting enzyme inhibitors in congestive heart failure. PMID- 3032813 TI - Hypothalamic-pituitary-adrenal interactions with the immune system. PMID- 3032815 TI - Delta hepatitis virus infection in China. AB - To assess the prevalence, epidemiological features and prognostic implications of hepatitis D (Delta) in Sichuan Province, The People's Republic of China, 649 sera (515 from HBsAg positive patients and 134 from HBsAg negative subjects) were tested by radioimmunoassay (RIA) for antibody to the hepatitis D virus (anti-HD). Forty-seven sera (7.2%) showed some degree of reactivity. Serial dilutions of these sera indicated that prozoning was not responsible for the equivocal results. Thirty-four of the 47 sera were submitted under code to a second laboratory for independent analysis. According to those results anti-HD antibodies were detected in four of these sera. The overall prevalence of anti-HD in the HBsAg positive patients therefore was 0.8% (4/515). On the basis of clinical, biochemical and histological data 427 HBsAg positive sera were further divided into acute Type B hepatitis, chronic Type B hepatitis, healthy carrier state and hepatocellular carcinoma (HCC) subgroups. Two of 65 (3.1%) anti-HD positive sera belonged to the acute Type B hepatitis group; one of 104 (0.9%), the chronic Type B hepatitis group and one of 246 (0.4%), the healthy carrier group. No antibody was detected in sera from 12 HBsAg positive HCC patients. All HBsAg negative patients were negative for anti-HD antibody. The results of this study indicate that despite a high prevalence of hepatitis B virus infection, positive serology for delta virus is uncommon in Sichuan Province, The People's Republic of China. PMID- 3032814 TI - A controlled trial of iodinated oil for the prevention of endemic cretinism: a long-term follow-up. AB - A double blind controlled trial designed to examine the effectiveness of intramuscular iodinated oil as a prophylactic for the nervous type of endemic cretinism was begun in 1966 in the highlands of Papua New Guinea. Infants born into the trial between 1966 and 1972 were followed up until 1982. The results showed that if the iodine supplement was given before conception the nervous form of endemic cretinism was prevented. Also a striking difference in the 15-year cumulative survival rate in favour of the test (iodinated oil) group was observed. Measures of motor and intellectual function revealed that children born to mothers given an iodine supplement performed significantly better. This observation shows that iodine deficiency leads to sub-clinical as well as clinical deficits. It also justifies the use of the term iodine deficiency disorder to cover the polymorphic nature of the abnormalities attributable to iodine deficiency. PMID- 3032816 TI - Comparative studies on aetiology and epidemiology of viral conjunctivitis in three countries of East Asia--Japan, Taiwan and South Korea. AB - A total number of 1105 cases with viral conjunctivitis, mainly epidemic kerato conjunctivitis (EKC) consisting of 354 cases from Sapporo, Japan, of 628 from Kaohsiung, Taiwan and of 123 from Busan, Korea, encountered during the periods, 1980-81 and/or 1983, were studied aetiologically and epidemiologically. Patients were aged from 27 days to 88 years with a peak in the 20-29 year age group. Aetiological diagnoses were established in 610 cases (55%), consisting of 536 cases (49%) caused by adenoviruses and of 74 (7%) by EV 70. The most frequently detected agent was Ad 8 (57%), followed by EV 70 (12%), Ad 3 (9%) and Ad 19 (7%). The aetiological profiles of viral conjunctivitis were generally similar in three cities of East Asia. EKC (70%) was mainly caused by Ad 8, Ad 19 and Ad 37, but AHC (13%) by EV 70 and PCF (5%) by Ad 3 and Ad 11. Ad 19 and Ad 37 isolates in three cities were compared with the cleavage patterns with restriction endonucleases, BamHl, Smal, Sall and Hindlll. All of the Ad 19 isolates tested were identical to Ad 19a reported by Wadell et al, and all but three isolates of Ad 37 were identical to the Ad 37 prototype prevalent in Europe and the US. The three isolates of Ad 37, different from the prototype in cleavage pattern with Hindlll, designated as Ad 37A, were detected in Sapporo and Kaohsiung in 1980. From the cleavage patterns with four restriction endonucleases, Hindlll, BamHl, Sa1l and Sstl, the Ad 8 isolates in 1983 were divided into three subtypes, which were associated with the cities isolated.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3032817 TI - Human papillomavirus type 16 in intraepithelial neoplasia (bowenoid papulosis) and coexistent invasive carcinoma of the vulva. AB - Tissues from two cases of bowenoid papulosis of the vulva with coexistent invasive squamous cell carcinoma were evaluated for the presence of human papillomaviruses (HPVs) and for nuclear DNA content. In both cases, HPV type 16 and nuclear aneuploidy were found in bowenoid papulosis as well as in invasive carcinoma. Patterns of hybridization suggested that the viral genome was integrated into the cellular genome in both bowenoid papulosis tissues as well as in invasive carcinoma tissues. These observations suggest that lesions designated as bowenoid papulosis may have invasive cancer potential. The term vulvar intraepithelial neoplasia seems to be more appropriate for HPV-16-containing aneuploid, intraepithelial lesions of the vulva. PMID- 3032818 TI - Abstracts of papers presented at the symposium of the International Society of Gynecological Pathologists. Vienna, Austria, September 5, 1986. PMID- 3032820 TI - Dietary fibre and obesity. Proceedings of a symposium at 5th International Congress on Obesity. 16 September 1986, Jerusalem, Israel. PMID- 3032819 TI - Studies on insulin secretion and the pituitary insulin secretagogue beta-cell tropin in the sand-rat (Psammomys obesus). AB - The sand-rat (Psammomys obesus) is an animal model for the study of human maturity onset diabetes which appears to be controlled by caloric intake. In the present investigations, these animals have been studied in relation to the influence of low- and high-energy diets on body weight, plasma insulin and blood glucose levels, and on insulin secretion from the perfused pancreas and the secretion of corticotropin-like intermediate lobe peptide (CLIP, ACTH18-39) and the insulin secretagogue beta-cell-tropin (beta-CT, ACTH22-39) from the pituitary neurointermediate lobe. The sand-rats maintained on the high-energy diet all became obese. Insulin secretion from the perfused pancreas of the obese sand-rat in the presence of 5.6 mM glucose was significantly higher than in the lean controls maintained on low-energy diets. Increasing the glucose concentration to 16.7 mM only produced a small stimulation of insulin secretion in the obese animals, and the difference between the two groups was not significant. Stimulation of insulin secretion by beta-CT was variable, but the obese animals appeared to be more responsive. Pituitary neurointermediate lobes were incubated for 4 h to measure the secretion of the ACTH related peptide. These were separated by gel filtration and the concentrations measured by radioimmunoassay with a CLIP antiserum and a CLIP standard. In all experiments beta-CT was 4-6 per cent of the total CLIP immunoreactive material. In these experiments the obese animals maintained on a high-energy diet were divided into two groups, those with plasma insulin levels less than 500 mu u/ml and those with insulin levels greater than 500 mu u/ml. The latter group had a significantly higher blood glucose level, presumably due to the insulin resistance resulting from the severe hyperinsulinaemia. It was also observed that CLIP-IRM and beta-CT secretion was lower in this group than in the animals maintained on low-energy diets or those on high-energy diets with moderate hyperinsulinaemia. This suggests a possible feedback inhibition by insulin on the secretion of beta-CT. PMID- 3032821 TI - Food energy values of dietary fibre components and decreased deposition of body fat. AB - The effects of guar gum (GG) and solka floc cellulose (SF) on food intake, stomach emptying and body fat deposition in rats were assessed together with their food energy values. Voluntary food intake in meal-fed rats (ca 100 g, male Wistar kept at 21C) was depressed initially by 40 per cent on adding 10 g GG to 100 g fibre free (FF) semisynthetic diet. This effect was not sustained, intake doubling within 15 d. Similar effects were not observed with SF. In pair-fed animals neither GG nor SF affected the rate of stomach emptying after 28 days. Digestible energy (DE) intake per rat over the 28 days was GG diet = 4616, SF diet = 4410, FF diet = 4373 kJ but body fat was lower in GG (25%) and SF (16%) fed rats than in FF fed rats. The calculated DE values of the fibre components were 10.1 kJ/g GG and 1.5 kJ/g SF (sem 1 kJ/g). After including body fat into the equations for metabolizable energy, the energy values were -7.1 kJ/g GG and -4.8 kJ/g SF (sem 2 kJ/g). This is equivalent to an increased energy expenditure of 17.2 kJ/g GG and 6.3 kJ/g SF. For GG an increased intestinal mucosal mass and cell turnover explains part of the apparent increase in energy demand. Our overall conclusion is that under certain circumstances and with regard to fat deposition some fibre components can be attributed negative energy values. PMID- 3032822 TI - Dietary fibre, diabetes and obesity. AB - An increased intake of dietary fibre appears to be useful for the treatment of both obesity and diabetes mellitus. Fibre-rich food is usually satisfying without being calorically dense. Supplementing a normal diet with gel-forming fibres, such as guar gum, leads to an increased satiation probably due to a slower gastric emptying. Recent long-term studies have confirmed the usefulness of viscous fibres as an adjunct to regular dietary treatment of obesity. Apart from a beneficial effect during caloric restriction, dietary fibre may improve some of the metabolic aberrations seen in obesity. Gel-forming fibres are particularly effective in reducing elevated LDL-cholesterol without changing the HDL-fraction. Impaired glucose tolerance or manifest diabetes is also improved. These effects are probably in part associated with the gelling property of the fibre which leads to an increased viscosity of the unstirred layer thereby delaying the absorption process. Other sources of dietary fibre with a high content of viscous gums, such as oats, have been shown to reduce LDL-cholesterol. Increased intake of viscous fibre leads to a gradual reduction in fasting glucose levels in diabetics. The reason for this is unclear but it cannot readily be explained by a delayed absorption process. Since insulin levels are also reduced these findings suggest that insulin resistance is alleviated. Recent studies with the euglycemic clamp technique support this possibility. Glucose uptake by isolated fat cells and both insulin sensitivity and responsiveness are also increased. PMID- 3032823 TI - Dietary fibre: mechanisms of action. AB - Actions of dietary fibre in the gut may be important in the control of energy intake and energy digestibility. These include effects on gastric emptying, upper small gut motility, rates and completeness of carbohydrate, fat and protein absorption in the gut. The ileal brake, whereby fat in the terminal ileum slows upper gut motility and inhibits ad-lib food intake, demands further investigation. Energy digestibility may be important, but large changes of fibre intake result in only small changes in digestibility. Gastrointestinal hormone release which may be affected by fibre is another topic for further investigation, and the effect of some types of fibre in reducing postprandial blood glucose and insulin levels, and thereby increasing insulin sensitivity, may be important for lipolysis and lipogenesis. PMID- 3032824 TI - Dietary fibre and lipid metabolism. AB - The influence of dietary fibre on lipid metabolism can be assessed at several levels. In animal studies it is possible to investigate the effects of fibre on serum and tissue lipid levels, on lipid absorption and excretion and, in specific instances, on experimental atherosclerosis. In man we are limited to examination of blood lipid and lipoprotein levels and on patterns of lipid excretion. Ershoff and his co-workers (eg) were among the first to demonstrate beneficial effects of fibre on serum and liver cholesterol levels in cholesterol-fed rats. They also showed that not all that we call fibre is hypocholesterolaemic. Cellulose, for instance, caused increases in liver cholesterol levels in rats. Cellulose also leads to increased levels of carcass cholesterol. Experiments in chickens, rabbits and monkeys have shown cellulose to exert the least beneficial effect on atherosclerosis or aortic sudanophilia. In man the insoluble fibres such as bran or cellulose are not hypocholesterolaemic, whereas soluble fibres such as guar gum or pectin are. The effects are also reflected in levels of high and low density lipoproteins. Studies relating to mechanism(s) of fibre action have shown that fibre may increase cholesterol and bile acid excretion and, in some cases, may affect faecal bile acid composition. Fibres have been shown to bind bile acids in vitro, the extent of binding being a function of both the type of fibre used and the type of bile acid or bile salt being studied. Fibre has also been shown to bind other lipids. Fibre also influences the rate of cholesterol absorption. To date most results can be classified according to the type of fibre used, ie soluble or insoluble; ionic or non-ionic. Our added understanding of modes of action of fibre will depend on better ability to relate structure to function. PMID- 3032825 TI - Dietary fibre, minerals and vitamins. AB - In Western populations a century ago diets consumed by the masses included a large amount of little refined cereal products. Intakes of vegetables (other than potatoes) and fruit, also foodstuffs of animal origin, tended to be low. Since then the situation changed conversely. There is now a third stage of change- authorities are urging reversion toward greater reliance on foods of plant origin, with increased intakes of less refined cereal products, legumes and vegetables and fruit. In developing countries, the first two stages are represented by rural traditional and urban partially Westernized diets. This paper provides details of intakes of diets generally, especially of dietary fibre, mineral salts and vitamins during the stages referred to, in both First and Third World populations, and discusses their bearing on the occurrences of nutrition-related diseases. PMID- 3032826 TI - New sources of dietary fibre. AB - Two sources of dietary fibre were discussed in this presentation: soybean and fenugreek. Soybean dietary fibre (SDF) was found to be effective in reducing plasma glucose levels in diabetic and fa/fa rats, ob/ob mice and in non-insulin dependent diabetes mellitus (NIDDM) subjects. Supplementation of SDF in bread was more effective in glucose reduction than powder. SDF was also found to be more effective in subjects with fasting blood glucose levels above 7.2 mmol/l. SDF had no effect on insulin levels in rats or NIDDM subjects although the insulin levels in ob/ob mice were lower after 180 d feeding. SDF had no effect on body weight or lipid levels in rats and human subjects. However, in diabetic rats with high levels of blood cholesterol, SDF feeding decreased the cholesterol levels after 45 d SDF administration. Addition of powdered fenugreek to an oral glucose tolerance test significantly reduced the subsequent postprandial blood glucose level in diabetic rats. Inclusion of fenugreek to the meal tolerance test given to NIDDM also decreased the postprandial blood glucose levels. Fenugreek was found to reduce the rate of gastric emptying and to inhibit glucose transport, indicating the blood modulating effect of fenugreek to be due mainly to delayed gastric emptying with direct interference with intestinal glucose absorption. Soybean and fenugreek dietary fibres reveal a potential benefit for the control of glucose metabolism in diabetes with additional advantages resulting from their ease in usage either in a mixture of water or milk products or in cooking. PMID- 3032827 TI - The influence of a high-fibre diet on body weight, serum lipids and blood pressure in slightly overweight persons. A randomized, double-blind, placebo controlled investigation with diet and fibre tablets (DumoVital). AB - Sixty slightly overweight women were treated with a weight-reducing diet for 12 weeks in a randomized, double-blind, placebo-controlled study. In addition to the diet 30 women received dietary fibre tablets, whereas the remaining 30 women received identical-looking placebo tablets. During the trial both groups experienced a significant reduction in body weight (P less than 0.01). The mean weight loss 8.5 kg (7.5-9.5 kg) in the fibre group was significantly higher than that of the placebo group 6.7 kg (4.8-8.0 kg) (P less than 0.01). Both serum triglyceride and serum cholesterol concentrations were significantly lowered (P less than or equal to 0.02) after treatment in both groups. No significant differences were detected between the groups. Both systolic and diastolic blood pressure were significantly reduced (P less than 0.01) in the fibre group. No significant reduction in blood pressure was found in the placebo group. Side effects, which were gastrointestinal in nature, were of low frequency. We conclude that supplementation with dietary fibre of the form used in this study is useful in the treatment of overweight women. PMID- 3032829 TI - Dietary treatment of obesity localized in different regions. The effect of dietary fibre on relapse. AB - Ninety-four obese women were matched in pairs with similar body mass index and age but with as wide variations as possible in regional obesity, estimated by the waist/hip (W/H) circumference ratio. Body composition and metabolic variables were determined. The women were instructed to lower their energy intake to 1100 kcal/d, and were then seen every 4th week by a dietitian. After 5 per cent weight decrease initial measurements of body composition were repeated and when reaching weight steady state (less than 2 kg weight decrease upon two visits) both body composition and metabolic variables were again determined. Before the start of diet there was a positive correlation between W/H ratio on the one hand and insulin and alanine aminotransferase on the other. There was no correlation between rate of weight decrease, loss of cell body mass or diminution of circumferences with the W/H ratio. The examination of potential effects of dietary fibre on relapse is currently in progress. PMID- 3032828 TI - Effects of a moderate dietary fibre supplement on hunger rating, energy input and faecal energy output in young, healthy volunteers. A randomized, double-blind, cross-over trial. AB - The effects of moderate dietary-fibre supplementation on satiety, energy intake and faecal energy excretion were studied in 20 young healthy volunteers of normal body weight, mean body mass index 20.9, receiving a dietary fibre supplement of 7.3 g per day in a randomized, double-blind, cross-over study. Hunger feeling, energy intake, and defaecation pattern, were recorded daily during a 2-week control period and then, during two 4-week treatment periods. Furthermore, faecal energy output was determined during the last week of each treatment period. The fibre treatment, as compared to placebo, resulted in a significantly higher faecal energy excretion: 173 kcal/d (163-183 kcal/d) vs 153 kcal/d (135-171 kcal/d), respectively (P less than 0.05); a decrease in hunger rating (using a visual analogue scale) (P less than 0.05); an increase in number of bowel movements (P less than 0.05), and a softer consistency of the stools (P less than 0.05). There was no significant difference in mean energy intake between the two treatment periods. This study demonstrated that moderate dietary fibre supplementation in normal man increases faecal energy excretion with simultaneously decreased hunger feeling. These beneficial effects may have therapeutic value in the management of obesity. PMID- 3032830 TI - The effect of high and low-fibre breakfasts on hunger, satiety and food intake in a subsequent meal. AB - Twenty normal weight female volunteers divided into high and low restraint groups consumed breakfast meals of high and low-fibre content (12.0 g and 3.0 g fibre respectively) on two separate occasions. Visual analogue scales were used to record hunger, fullness, desire to eat, and a measure of prospective consumption for 21/2 hours after each meal. At this point, a tray of pre-weighed lunch foods was offered and subjects were requested to eat as much or as little as they desired. The two breakfast meals (of equal weight) were based on toast, breakfast cereal, milk, butter and orange marmalade. No significant difference in energy intake at lunch was found after the high and low fibre breakfasts, or between the restraint groups. There was no significant difference between ratings after the high and low-fibre meals except for fullness, which was greater after the high fibre breakfast. The effect of fibre overall, was relatively weak compared to the differences between the two restraint groups, with the high restraint group consistently expressing significantly less hunger before, during and after the breakfasts compared to the low restraint group. PMID- 3032831 TI - Satiation, satiety and the action of fibre on food intake. AB - This paper sets out to provide a review of the effect of fibre upon the motivation to eat and ultimately upon the amount of energy consumed. In addition the authors suggest various ways in which the analysis and understanding of the effects of fibre may be extended by embracing different types of experimental design and varying the assessment procedures. In the course of this work it became obvious that some important pieces of information had yet to be demonstrated. In particular it seems necessary for studies to measure the actual amounts of energy ingested in addition to tapping motivational indices. Moreover, the evaluation of the motivation to eat could become more sensitive and more comprehensive. Evidence suggests that fibre does exert effects on the short-term control of food consumption. The action appears to be expressed both within and between meals. Both of these effects may be clinically relevant. It has yet to be determined the extent to which these effects are influenced by the mode of administration of the fibre, the type of fibre and the amount. PMID- 3032832 TI - Effect of dietary fibre on postprandial thermogenesis. AB - The effect of fibre on postprandial thermogenesis was evaluated in seven healthy males (age 29.6 +/- 3.6 years, BMI 24.6 +/- 0.6 kg/m2) after they had ingested three isocaloric meals with different fibre contents: low fibre (Lfb) containing 8 g. dietary fibre; glucomannan (Glc), which was Lfb supplemented with 6 g glucomannan, a pectin-like fibre; and high fibre (Hfb) containing 26 g dietary fibre. Postprandial thermogenesis, evaluated for 6 h after the ingestion of food, was higher after the Lfb (82.3 +/- 5.4 kcal) than after the Hfb and Glc meals (69.4 +/- 6.8 and 61.4 +/- 8.4 kcal, respectively). Glucose and insulin responses were depressed over the first 2 h after the Hfb and Glc meals as compared to the Lfb meal. However, these differences disappeared when the whole 6 h postprandial period was considered. No major effects of the different fibre contents of the meals on postprandial triglyceride and FFA levels were detected. These results demonstrate that the fibre content of meals, besides the well known effect on energy intake, may also affect energy expenditure. PMID- 3032833 TI - In vitro study on humoral encapsulation of microfilariae: effects of diethyldithiocarbamate and dopachrome on the reaction. PMID- 3032834 TI - Amino acid sequence of UP1, an hnRNP-derived single-stranded nucleic acid binding protein from calf thymus. AB - The UP1 single-stranded nucleic acid binding protein from calf thymus (Herrick, G. & Alberts, B.M. (1976) J. Biol. Chem. 251, 2124-2132) has recently been shown to be a proteolytic fragment derived from the A1 heterogeneous nuclear ribonucleoprotein (hnRNP) (Pandolfo et al. (1985) Nucleic Acids Res. 13, 6577 6590). The NH2-terminus of the 22,162 dalton UP1 protein appears to be blocked, which suggests that UP1 represents the NH2-terminal two thirds of this 32,000 dalton hnRNP protein. The complete amino acid sequence for UP1 was derived from automated sequencing of peptides that were purified by HPLC from digests with trypsin, chymotrypsin, Staphylococcus aureus protease, endoproteinase Lys-C, and cyanogen bromide. Trichloroacetic acid precipitation followed by enzymatic digestion in 2 M urea proved to be the best approach for generating UP1 peptides. By carboxymethylating after, rather than before, digestion it was possible to avoid problems associated with the insolubility of the carboxymethylated UP1. All of the resulting peptides in amounts varying from 2 to 15 nmol were coupled to aminopolystyrene prior to solid-phase sequencing. Using these methods, no difficulties were encountered in assigning glutamic acid residues or in completely sequencing peptides that contained up to 25-30 residues. The relative ease with which the UP1 protein was sequenced, requiring only about a year to complete, and the comparatively modest amount of protein required, less than 5 mg, attests to the usefulness of water soluble carbodiimide coupling and solid phase sequencing for determining the primary structures of proteins. In addition to serving as a basis for determining structural relationships among various mammalian single-stranded nucleic acid binding proteins, the amino acid sequence of UP1 reveals that the A1 hnRNP protein contains a region of internal sequence homology that apparently corresponds to two independent nucleic acid binding sites. PMID- 3032835 TI - Degree of chromatin fragmentation and frequency of nuclear pyknosis in Percoll fractionated thymocytes of irradiated rats. AB - The relationship between nuclear chromatin degradation to polydeoxyribonucleotides (PDN) and other features of interphase death were studied using thymocytes of normal and X-irradiated rats. Fractionation of the thymic cells in Percoll gradients was performed in order to separate dead from intact cells. The degree of radiation-induced chromatin fragmentation, as assessed by electrophoresis, was similar for PDN from all Percoll bands. Following irradiation 87-98 per cent of 'heavy' thymocytes were pyknotic and almost devoid of receptors to autologous erythrocytes thus comprising a dead cell population. A direct relationship between PDN content and nuclear pyknosis was noted throughout the gradient. The loss of autologous rosette-forming ability was directly related to other indices of interphase death. The possibility of PDN originating from pyknosis-prone cells and the capacity of radiosensitive thymocytes to form autologous rosettes are discussed. PMID- 3032836 TI - [Secondary achalasia within the scope of polyneuropathy in small cell bronchial cancer]. PMID- 3032837 TI - Beta-adrenergic receptors in human trabecular meshwork. Identification and autoradiographic localization. AB - Beta-adrenergic receptors were identified in slide-mounted sections of human trabecular meshwork by in-vitro labeling, light microscopic autoradiography. Autoradiograms were generated after incubation of slide-mounted tissue sections with 125I-cyanopindolol, a selective high-affinity probe for beta-adrenergic receptors. Experiments in which an excess of either unlabeled Practolol (beta 1 selective ligand) or Zinterol (beta 2 selective ligand) were included suggested that most of the receptors were of the beta 2 subtype. Data from displacement studies using increasing concentrations of the highly specific beta 1- and beta 2 adrenergic receptor antagonists, ICI-89,406 and ICI-118,551, respectively, confirmed the predominance of the beta 2-adrenergic receptors. Similar results obtained with cultured trabecular endothelial cells suggest that the beta adrenergic receptors may be associated with the endothelial cells of the trabecular meshwork in vivo. These results provide anatomic evidence for the hypothesis that beta-adrenergic agents improve outflow by direct action on the trabecular meshwork and provide a rationale for the development of more selective beta 2-adrenergic agents to increase outflow facility. PMID- 3032838 TI - Regulation of lens cyclic nucleotide metabolism by Ca2+ plus calmodulin. AB - Adenylate cyclase activity was identified in membranes isolated from bovine lens fiber cells. Basal activity, in the presence of microM Ca2+ was stimulated by either sodium fluoride, guanosine 5'-[alpha,beta-imido]triphosphate (Gpp(NH)p), or forskolin; ethylene glycolbis(2-aminoethylether) tetraacetic acid (EGTA) markedly inhibited both the basal activity and the extent of stimulation by these agents. Exogenous calmodulin enhanced the Ca2+-dependent stimulation of adenylate cyclase activity. In the presence of optimal concentrations of Ca2+ plus calmodulin, adenylate cyclase activity was approximately 15 times greater than that in the presence of EGTA. Adenylate cyclase activity was not stimulated by a number of potential agonists that included carbachol, serotonin, prostaglandin E1 (PGE1), prostaglandin E2 (PGE2), adenosine, isoproterenol epinephrine, dopamine, and phenylephrine. The presence of the Ns and Ni guanine nucleotide regulatory complexes was indicated by two observations: Cholera toxin catalyzed the adenosine diphosphate (ADP) ribosylation of a number of lens membrane proteins, including a 46,500-dalton component (likely the alpha-subunit of Ns), and Pertussis toxin catalyzed the ADP ribosylation of a single 41,000-dalton lens membrane component (likely the alpha-subunit of Ni). However, that Gpp(NH)p did not inhibit either the forskolin-activated or the calmodulin-activated adenylate cyclase activities does not indicate a role for Ni in regulating this enzyme. Both cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) phosphodiesterase activities were identified in a supernate fraction derived from bovine lens. The cAMP phosphodiesterase activity appeared to be predominantly the low Km form of the enzyme. The cGMP phosphodiesterase activity, which was Ca2+-dependent, was partly inhibited maximally by 7 microM R24571, indicating its probable calmodulin dependence.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3032839 TI - Retinitis and deviant immune responses following intravitreal inoculation of HSV 1. AB - Following anterior chamber inoculation of herpes simplex virus type 1 (HSV-1) into one eye of adult, immunocompetent BALB/c mice, an interesting pattern of ocular pathology and systemic immune responses emerges, characterized by destruction of the contralateral retina with sparing of the ipsilateral retina; and impairment of delayed hypersensitivity (DH) accompanied by intact humoral immunity to the virus. Experiments using animals inoculated via the intravitreal route revealed a different pattern of retinitis in the inoculated and in the uninoculated fellow eye following intravitreal inoculation of HSV-1. The retinitis in the inoculated eye results in localized, focal necrosis with concomitant preservation of the retinal architecture in areas juxtaposed to those in which retinal destruction occurs. The retinitis in the uninoculated contralateral eye is characterized by pan-retinal inflammation and subsequent loss of the architecture of the entire retina. Intravitreally inoculated animals exhibited virus-specific impairment of DH responses to HSV-1 but were capable of producing anti-HSV-1 neutralizing antibody. Impairment of DH response after inoculation of live HSV-1 suggests that intraocular processing of viral antigens occurs such that processed antigens are released systemically in a soluble form. PMID- 3032841 TI - Human papillomavirus type 38 isolated from patients with epidermodysplasia verruciformis. AB - A new type of human papillomavirus (HPV), HPV-38b, was isolated from patients with epidermodysplasia verruciformis and was molecularly cloned. This HPV was shown by hybridization experiments to have almost no sequence homology with other types of reported HPVs, but did show homology with HPV-38, which was recently isolated from a melanoma. A physical map and a rough genetic map of the organization of this HPV are presented. PMID- 3032840 TI - Clearance and localization of intravitreal liposomes in the aphakic vitrectomized eye. AB - The authors have examined the fate of intravitreally injected liposomes in the aphakic, vitrectomized eye of the rabbit. Liposomes labelled with 125[I]-p hydroxybenzimidylphosphatidylethanolamine were eliminated rapidly from the intraocular fluid. Nonetheless, a significant fraction of these liposomes were found to bind to various ocular tissues including the retina, iris, sclera, and cornea. Ultrastructural studies with gold colloid-loaded liposomes revealed that retinal bound liposomes were attached to the inner limiting lamina but did not penetrate to the internal cells of the retina. Epiretinal cells bound and internalized gold colloid-loaded liposomes suggesting that these cells may be very sensitive to liposome mediated drug delivery. PMID- 3032843 TI - Host involvement in vaccinia virus replication. AB - Nutrient-depleted conditions, which induced an inhibited state in uninfected cell monolayers, rendered the cells nonpermissive for vaccinia virus, while not affecting their ability to support propagation of an unrelated mengovirus. When the inhibited state was reversed, the ability to propagate vaccinia virus was restored. The inhibited cells retained the ability to support some early-induced events (thymidine kinase synthesis and its arrest, which are DNA synthesis independent functions in a mouse cell host), but lost the ability to support some others (sensitization of input viral DNA to DNase, induction of DNA polymerase synthesis, and virus-induced CPE). The data are consistent with involvement of a host factor(s) in induction of early vaccinia virus-induced events in infected cells. PMID- 3032842 TI - Comparative analysis of the canAV-1 and canAV-2 genomes. AB - The molecular weights of canine adenovirus type 1 (canAV-1, strain Utrecht) and type 2 (canAV-2, strain Toronto A26/61) DNAs were determined by contour length measurements and restriction endonuclease analysis. In each case, an average molecular weight value of 20 (+/- 0.5) X 10(6) daltons was obtained with both methods. Similar in size, the canAV-1 and canAV-2 DNA molecules were, however, cleaved at very distinct positions by a variety of restriction endonucleases. The observed lack of DNA sequence homology confirms the existence of at least two different canine adenovirus species [Intervirology 23: 23-28 (1985)]. PMID- 3032844 TI - Zoster sine herpete causing encephalomyelitis. AB - The case of a patient with encephalomyelitis and laboratory signs of a central nervous system herpes zoster infection without cutaneous lesions is reported. The diagnosis was supported by the serological evidence of intrathecal synthesis of specific antibodies against Varicella-zoster virus (VZV). PMID- 3032845 TI - Pathophysiology of the neuromuscular junction in diabetic rats. AB - Using electrophysiological methods, we examined transport and transmission properties of the neuromuscular synapse in normal and diabetic rats. The stereospecific glucose transport system in the presynaptic nerve terminal was examined in two diabetic models and compared to the control. In both the streptozotocin (STZ) model and the Cohen model, a significant reduction was observed in the rates of glucose transport. A semiquantitative index for glucose transport, the glucose uptake factor, was 61 +/- 18% (SD) in the control group, 17 +/- 9% in the STZ group, and 15 +/- 10% in the Cohen group. Diabetes also affects the liberation of acetylcholine from the presynaptic motor nerve endings. The mean number of acetylcholine quanta liberated by the nerve impulse [quantal content (QC)] was reduced by 43% after 3.5 months of STZ diabetes. Thus, both the metabolic and the transmission functions of the neuromuscular junction are affected by the diabetic state. PMID- 3032847 TI - Modulation of synaptic potentials at central and peripheral synapses. PMID- 3032846 TI - The role of interstitial potassium in the generation of low-calcium hippocampal seizures. AB - The explosive nature of a focal cortical seizure suggests the operation of strong positive feedback in the neuronal network. It was previously proposed that in the hippocampus this may be provided by the regenerative accumulation of potassium in the interstitium. This hypothesis was severely criticized in the past. However, it gained new impetus with the recent discovery that focal seizures can arise in mammalian hippocampal slices perfused with low calcium solutions despite the block of chemical synaptic transmission. Here, we examine the relationship of interstitial potassium concentration to the electrogenesis of these low-calcium seizures. Both the experimental data and the behavior of a simplified mathematical model describing neuronal discharge in low calcium, support the contention that interstitial potassium accumulation may play an important role in the buildup of hippocampal seizures. PMID- 3032848 TI - Minimal brain damage induced by early iron deficiency: modified dopaminergic neurotransmission. AB - The reports that iron-deficiency anemia in human subjects induces behavioral changes was investigated in rats made nutritionally iron-deficient. The most prominent features of these animals were: the unchanged metabolism of the neurotransmitters noradrenaline, dopamine and serotonin, profound reduction of brain nonheme iron, the selective diminution of dopamine D2 receptor number (measured by Bmax), modification of dopamine-dependent behaviors and reduction of learning processes. The induction of these changes and their recovery with iron supplementation are age- and time-dependent phenomena. In newborn rats, however, the consequences of iron deficiency are irreversible, even after long-term iron supplementation. The results point to the profound effect iron metabolism can have on the long-term development and function of dopaminergic neurotransmission. These findings may not be totally unexpected, since iron distribution in the brain is highly localized in dopaminergic-peptidergic regions, such as the globus pallidus, substantia nigra, red nucleus, thalamus, caudate nucleus and nucleus accumbens. In some regions its concentration is higher than that found in the liver, the site of iron metabolism. PMID- 3032849 TI - Possible involvement of adenylate cyclase in learning and short-term memory. Experimental data and some theoretical considerations. AB - Two experimental approaches--the cellular and the neurogenetic--to the study of molecular mechanisms of elementary memory implicate the cyclic AMP cascade in general, and adenylate cyclase in particular, in the processes of acquisition and short-term memory. Models of learning and memory should account for four basic phenomena: persistence of memory, stimulus convergence, temporal specificity and memory decay. These phenomena thus place constraints on the structure and operation of any postulated memory apparatus. The relevant experimental data derived from the study of memory mutants in Drosophila and of the gill and siphon withdrawal reflex in Aplysia are discussed and analyzed in light of the above mentioned constraints. PMID- 3032850 TI - The role of endogenous opioid peptides in physiological and pharmacological reward responses--a survey of present-day knowledge. AB - Endogenous opioid peptides (EOP) participate in a variety of physiological and pharmacological responses that can be recognized by: increased concentrations of EOP in the cerebrospinal fluid; a combination of euphoria and analgesia; and suppression of the responses by specific opiate antagonists. Application of these responses is exemplified in acupuncture, in self-stimulation via hypothalamic electrodes, and in food and fluid consumption. For example, intake of sweet solutions exerts a biphasic effect: in the beginning pain threshold is elevated, but later on tolerance to morphine-induced analgesia develops. A supply of 2% NaCl also stimulates fluid intake as well as raising the pain threshold. Stress induced analgesia is due to coupled release of ACTH and beta-endorphin from the pituitary. In man, large variations in pain sensitivity may be related to the level of intrinsic EOP. PMID- 3032851 TI - Regulation of motor axon sprouting. AB - Our studies on the amphibian and mammalian motor systems suggest that sprouting of intact motoneurons and synapse formation can be regulated by three mechanisms: peripheral, central, and transneuronal. Peripheral mechanisms provide the means of a direct mode of interaction between the periphery of the nerve cell and the target, to determine the extent of target innervation. The central mechanism enables target muscles to signal the cell bodies of their innervating motoneurons to regulate axonal growth and synapse formation, and thus again determine the extent of their innervation. The transneuronal mechanism provides a vehicle by which the pattern of innervation of a muscle can be altered by nerve cells that do not themselves innervate the muscle, but are an integral part of the entire system. PMID- 3032852 TI - Inhibition of human polymorphonuclear leukocyte functions by ibuprofen. AB - We have found that pretreatment of human neutrophils with ibuprofen (0.10-1.0 mg/ml) results in an irreversible, concentration-dependent inhibition of superoxide anion generation and release of lysosomal enzymes (myeloperoxidase, lysozyme) stimulated by the synthetic peptide, N-formyl-methionyl-leucyl phenylalanine (FMLP), the complement fragment C5a, and to a lesser extent by serum opsonized zymosan. Inhibition of granule exocytosis and oxygen radical generation at ibuprofen concentrations less than 5 mg/ml was not due to drug cytotoxicity since release of the cytoplasmic enzyme lactate dehydrogenase was not affected by ibuprofen. In contrast to neutrophil responses mediated by C5a or FMLP, ibuprofen did not inhibit either enzyme release or superoxide anion generation by neutrophils stimulated with phorbol myristate acetate. Ibuprofen did not function as an oxygen radical scavenger in a cell-free system in which superoxide anion was generated by the aerobic action of xanthine oxidase on hypoxanthine. Ibuprofen also inhibited in a concentration-dependent fashion both directed migration (chemotaxis) and stimulated random migration (chemokinesis) of neutrophils exposed to either FMLP or C5a. Inhibition of neutrophil adherence to plastic surfaces and bovine pulmonary artery endothelial cells was equally effective when the neutrophils were treated with ibuprofen before stimulation with FMLP or phorbol myristate acetate. The inhibitory effects of ibuprofen pretreatment of neutrophils could not be overcome by addition of prostaglandins E1 or E2 (0.3-300 nM). These results demonstrate that ibuprofen is capable of suppressing many functions thought to be important in neutrophil-mediated acute pulmonary inflammatory processes. Results of these experiments further suggest that ibuprofen may inhibit neutrophil functions by acting on cellular components separate from membrane receptors or by blockade of cyclo-oxygenase products which may be involved in these neutrophil functions. PMID- 3032854 TI - Epstein-Barr virus (EBV) infection. PMID- 3032853 TI - Suppression of human lymphocyte responsiveness by forskolin: reversal by 12-O tetradecanoyl phorbol 13-acetate, diacylglycerol and ionomycin. AB - Forskolin, a potent activator of adenylate cyclase, was examined for its ability to alter human peripheral blood lymphocyte (HPBL) activation by both mitogens and antigens. We found that forskolin, at concentrations ranging from 0.04 to 25 micrograms/ml, caused a dose-dependent inhibition of HPBL responses to mitogens (concanavalin A, phytohemagglutinin, pokeweed mitogen and Staphylococcus aureus) and to recall antigens (tetanus toxoid and streptokinase/streptodornase). Inhibition was reflected in altered DNA, RNA and protein synthesis, including immunoglobulin production, and was not due to altered cell viability. Forskolin also induced a 19-fold increase in HPBL cyclic AMP levels at the same concentrations that suppressed HPBL function. To further define the mechanism(s) by which these elevations in cyclic AMP suppressed HPBL function, we tried to reverse these inhibitory effects with several agents; ascorbic acid, carbachol and levamisole had no effect. However, the phorbol ester, 12-O-tetradecanoyl phorbol 13-acetate, as well as L-alpha-1,2-dioleoyl diacylglycerol were able to completely reverse the inhibition. Furthermore, the Ca2+ ionophore, ionomycin, was also able to act synergistically with lower and less effective concentrations of 12-O-tetradecanoyl phorbol 13-acetate to reverse the inhibitory effects of forskolin. The data suggest that forskolin-induced elevations in cyclic AMP may lead to inhibition (or, more correctly, prevents the activation) of protein kinase C, presumably by inhibiting phospholipid turnover. Our studies suggest a linkage between these two opposing membrane-signal transduction systems with protein kinase C representing a pivotal point for various regulatory signals that ultimately control lymphocyte activation and function. PMID- 3032855 TI - Quantitative risk assessment of lung cancer in U.S. uranium miners. AB - The National Institute for Occupational Safety and Health (NIOSH) has recently updated the vital status of the U.S. cohort of U miners through the end of 1982. This represents 69 additional lung cancer deaths since the last published follow up through 1977. This more recent data was used to generate quantitative risk estimates of lung cancer after exposure to Rn daughters. Relative risks were estimated through use of the Cox proportional hazards model with an internal referent group. Results indicated that the exposure-response relationship was a slightly convex curve, predicting excess relative risks between 0.9 and 1.4 per 100 working level months (WLM) in the lower cumulative exposure range. Other findings of interest include a significant exposure-rate effect with low exposure rates more harmful per unit of cumulative exposure (WLM). Two temporal effects which modify relative risk estimates were also found. Relative risk increased with age at initial exposure to underground U mining. However, relative risk of lung cancer fell dramatically in the years following cessation of exposure. PMID- 3032856 TI - Optimizing the gross alpha counting method for determining Rn progeny levels in the atmosphere. PMID- 3032857 TI - Comment on 'An epidemiological analysis of the relationship between exposure to Rn progeny, smoking and bronchogenic carcinoma'. PMID- 3032858 TI - The influence of the charge and of the tin-ligand ratio on bone uptake of technetium 99m labelled osteotropic radiopharmaceuticals. AB - This work describes the determination of the charge for various 99mTc-Sn phosphonate complexes. The variation of the Sn/P ratio using an HMDP ligand produces complexes with different negative charges. Biological distribution studies show superior bone uptake of strongly negative charged complexes. PMID- 3032859 TI - Molecular weights of technetium hydroxyethylidene diphosphonate complexes. AB - The results of recycle size-exclusion and anion-exchange chromatography are combined to determine molecular weights and radii of gyration for selected components of a carrier-added technetium hydroxyethylidene diphosphonate radiopharmaceutical bone-imaging preparation. Samples evaluated by size-exclusion are selected from fractions collected as the preparation elutes from the anion exchange column. Polyethylene glycol and polyphosphate molecular weight standards are used, in conjunction with the universal calibration method, to determine Mark Houwink constants and to estimate molecular weights for the complexes. The results of this study support a spherical shape model for the collection of technetium complexes. Given the combined knowledge of molecular weights, charges, technetium concentration dependencies, and mass spectral data, several formula assignments are proposed, including TcO(OH)L-1, Tc2O(OH)6L-3, Tc2O(OH)2L3(-6), Tc3O2(OH)3L2(-5), Tc3O2(OH)3L3(-7), Tc2O2(OH)4(HL)3(-7) and Tc2O2(OH)4(H2L)4(-8). PMID- 3032860 TI - Sum peak method applied to the study of human platelet cells. AB - The sum peak method based on the phenomenon of perturbed angular correlation was applied to study the chemical environments of 111In in intact platelet cells. The sum peak appearing in the gamma-ray spectrum of 111In which emits two gamma-rays in cascade was observed for 111InCl3 solutions and 111In-labeled platelets. An increase in intensity ratios of the sum peak to its single peak was found when 111In incorporated into platelets from its aqueous solution. This fact was discussed in terms of physico-chemical environments of 111In together with an evaluation of sum peak analysis in the spectra. PMID- 3032861 TI - Cyclotron production of no-carrier-added 206Bi (6.24 d) and 205Bi (15.31 d) as tracers for biological studies and for the development of alpha-emitting radiotherapeutic agents. AB - The production and radiochemical purification of no-carrier-added (NCA) bismuth isotopes (i.e. 6.24 d 206Bi and 15.31 d 205Bi) was studied. The Bi isotopes are intended for use as tracers in biodistribution studies and in the design and testing of alpha-emitting radiotherapeutic agents. The total cross sections and yields for the production of 206Bi and 205Bi from the proton bombardment of natural Pb targets were measured using the stacked-foils technique. A radiochemical method for the separation and purification of NCA Bi radioactivities from a multi-gram Pb target was also tested, modified, and improved to provide aqueous solutions of chemically and biologically useful amounts of NCA 206,205Bi for measuring tissue and subcellular-distributions, and for studies covering several chemical aspects of the development of Pb/Bi radiotherapeutic agents. PMID- 3032862 TI - Radioprotection of sheep erythrocytes by polyvinylpyrrolidone. AB - The protective effect of polyvinylpyrrolidone of mol. wt 10,000 and 40,000, on sheep erythrocytes irradiated up to 13 kGy, and stored for one month at 4 degrees C was studied. Addition of these compounds at a level of 10-15% reduced hemolysis from 90% in irradiated erythrocyte solution to approximately 10%. The high efficiency of this protective agent is discussed. PMID- 3032863 TI - Targetry and radiochemical methods for the simultaneous cyclotron production of no-carrier-added radiopharmaceutical-quality 100Pd, 97Ru and 101mRh. AB - Target-dissolution and radiochemical methods were investigated to optimize the simultaneous production of radiopharmaceutical-quality, no-carrier-added (NCA) 3.63-d 100Pd, 2.88-d 97Ru and 4.26-d 101mRh. These radionuclides have potential as radiotracer labels and/or as short-range dose emitters for use with specific function radiopharmaceuticals being investigated for radioimmunotherapy applications. Metallic Rh (100% 103Rh) and RhCl3 X 3H2O were used as target materials. After bombardment with high energy protons these targets were subjected to a combination of procedures (i.e. electrolytic dissolution, ion exchange, and solvent extraction) in order to separate the desired radionuclides. The use of a single cyclotron target in combination with several radiochemical processes were investigated to simultaneously produce these radionuclides in high specific activities and radiochemical forms suitable for radiopharmaceutical syntheses. PMID- 3032864 TI - Comparative evaluation of electrophilic aromatic iododemetallation techniques for labeling radiopharmaceuticals with iodine-122. AB - A series of metallated arenes bearing sigma-bonded tin, mercury, germanium, boron, or silicon were evaluated as labeling substrates for the generator produced positron emitter 122I(t1/2 = 3.6 min). Using dichloramine-T with 122I, radiochemical yields exceeding 90% were achieved after 1 min at 25 degrees C using stannylated or mercurated arenes in ethanol, or germylated arenes in acidic solvent. Stannylated or mercurated arenes resulted in high labeling yields even with deactivated aromatics in ethanol, but regiospecifically-iodinated products were obtained only from stannylated precursors due to differences in the syntheses of these organometallics. The implications of these results to the labeling of radiopharmaceuticals with 122I are discussed. PMID- 3032865 TI - The effect of temperature of fluorescence for liquid scintillators and their solvents. AB - The pulse-height distributions for 241Am and 131mXe in PPO solutions of the aromatic hydrocarbons, i.e. benzene, o-xylene, m-xylene, ethylbenzene and cumene are found to shift toward higher pulse-heights with decreasing temperature. Under u.v. excitation, the fluorescence intensity of these pure aromatic hydrocarbons increases markedly and the fluorescence maximum is found to shift to longer wavelengths with decreasing temperature. In conjunction with observations of the pulse-height shift in liquid scintillators and the fluorescence emission from the pure solvents, the pulse-height shifts observed are interpreted in terms of excimer formation in the aromatic hydrocarbons. PMID- 3032866 TI - Optimization of [11C]HCN production and no-carrier-added [1-11C]amino acid synthesis. AB - The optimal conditions for the catalytic production of [11C]HCN from [11C]CO2 were investigated. [11C]CO2 was reduced to [11C]CH4 with H2 on Ni and then converted to [11C]HCN by reaction with NH3 on Pt in a radiochemical yield of more than 95% under the optimized conditions of an NH3 concentration of 5 vol%, a Pt furnace temperature of 920 degrees C, and a reaction gas flow rate of over 200 mL/min. Absorbers were used to remove O2 and H2O from the reaction gas. The synthesis of no-carrier-added [1-11C]amino acids from [11C]HCN via [11C]aminonitriles was successfully carried out. This method is suitable for automation of [1-11C]amino acid production. PMID- 3032867 TI - [Cytomegalovirus infections following heart surgery. A prospective study of specific antibodies]. PMID- 3032868 TI - [Brain metastasis as the initial manifestation of bronchial carcinoma: value of combination surgery of the metastasis and the primary tumor]. PMID- 3032869 TI - [Rotavirus infections in childhood: studies using a molecular biology method (gene electrophoresis)]. AB - During a 15-month period all children below 16 years admitted to the Children's Hospital in St. Gallen with acute diarrheal disease were studied for rotavirus (RV) infection. Stool samples from control patients without gastrointestinal disease were investigated for RV shedding in order to detect RV carriers or asymptomatically infected children. RV was detected by electrophoresis of RV genomic dsRNA in 154 children. 119 (58%) of 205 patients hospitalized because of diarrhea were RV associated, 25 children became symptomatically ill during their hospitalization, and in 10 (1%) of 954 control patients RV could be found. During the winter 1983/1984 two epidemic peaks of RV associated diarrhea were noted, whereas in the summer period RV related disease occurred only sporadically. The peak incidence of RV infection was in the age group between 10 and 12 months. After the 3rd year of life RV infection only rarely required hospitalization. During this study period 8 different genomic RNA patterns of RV were found. The number of patients within these 8 RV types, however, is too small to allow definite correlations between epidemiological or clinical features and selected electrophorotypes. The possibility to perform refined epidemiological and clinical analyses of RV infection by genomic dsRNA electrophoresis offers important advantages when compared to other RV detection systems. In addition, this method has proven to be fast, simple and reliable. PMID- 3032870 TI - [Possibilities and limits in the detection of nosocomial rotavirus infections]. AB - During an epidemiological survey on rotavirus gastroenteritis in the area of Berne (Switzerland) different virus types were analyzed according to their genome segment pattern. Nosocomial rotavirus infections among pediatric patients were carefully investigated. Possible limitations of such studies are discussed in details. PMID- 3032871 TI - Mitochondrial DNA insertion polymorphism and germ line heteroplasmy in the Triturus cristatus complex. AB - Restriction enzyme analysis of mitochondrial (mt) DNA isolated from oocytes of 185 individuals of the T. cristatus complex collected from 10 European countries has demonstrated that large length variation (greater than 40 bp) is a common feature of the group. Insertion polymorphism was found both within and among populations, and in all cases maps to the control region of the molecule. In addition, 2 individuals from Pisa, (Italy) were each found to be heteroplasmic for 2 large insertions comprising tandem repeats of 1100 bp of the control region. Large-scale length variation has been described in a few other lower vertebrates, but some of the insertion variants within populations described here are of unprecedented size (up to 8500 bp). This is in dramatic contrast to mammalian mtDNA in which size variation is largely restricted to small (less than 15 bp) insertions and deletions. PMID- 3032872 TI - A high-fibre diet in gestational diabetes--wheat fibre, leguminous fibre or both? PMID- 3032873 TI - Immunocytochemical demonstration of proopiomelanocortin- and other opioid-related substances and a CRF-like peptide in the gut of the american cockroach, Periplaneta americana L. AB - Using the peroxidase-antiperoxidase technique, we showed the presence of peptides which are immunologically resembling mammalian corticotropin releasing hormone (CRF)-, adrenocorticotropic hormone (ACTH)-, beta-endorphin (beta-END)-, alpha melanocyte stimulating hormone (alpha-MSH)-, methionine-enkephalin (met-ENK)- and leucine enkephalin (leu-ENK)- like immunoreactivity in hundreds to thousands of endocrine cells and nerve fibers in the midgut of the American cockroach Periplaneta americana. In the cockroach hindgut no immunoreactive cell bodies could be observed, although nerve fibers were clearly noticed to be recognized by antisera to CRF, ACTH1-24, ACTH11-24 and beta-END. Nothing is exactly known as to the function(s) of the demonstrated materials, but one can speculate that these numerous immunoreactive cells, might have important paracrine and/or endocrine functions in the insect physiology. PMID- 3032874 TI - Importance of HLA-DQ and -DP restriction elements in T-cell responses to soluble antigens: mutational analysis. AB - We describe a new approach to delineating the restriction elements used by antigen-specific human T-cell lines. EBV-transformed B cell lines and congenic HLA class II antigen-loss mutants are used to present soluble antigen to immune T cells. In this way it is possible to assess the independent contribution of individual class II loci to the restriction repertoire of the T cells. In contrast to results obtained with other methods of restriction element analysis, we find that approximately 40% of the T-cell response to several antigens is restricted by non-DR class II molecules. Both mutational analysis and blocking by class II specific monoclonal antibodies demonstrate that the non-DR restricted responses derive from DQ and DP-encoded determinants. We also find specific DR/DQ haplotype preferences for the presentation of some but not all antigens. Using a mutant that expresses only the DQ1 molecule, and is derived from a DR1, DQ1 parent line, we demonstrate a functional split of serologically defined DQ1 molecules consistent with the electrophoretic variation reported between DQ1 molecules linked to DR1 and those linked to DR2. Pairs of mutants that differ by expression of a single class II protein reveal a much broader use of available class II restriction elements than previously recognized. PMID- 3032876 TI - Cervico-facial adenoid cystic carcinoma: study of 102 cases. Influence of radiation therapy. AB - One hundred two patients with cervico-facial adenoid cystic carcinoma were treated with surgery alone, radiotherapy alone or both from 1951 to 1980. All the cases have a 5-year minimum follow-up. The local control rate is 55.5% at 5 years and 37.7% at 10 years. The 5-year local control rate is 44% with surgery alone, 65.8% with radiotherapy alone and 77.8% with post-operative radiotherapy. The difference between surgery alone and radiosurgical association is significant (p less than 0.01). The bone involvement diminished local control rate (32.2%/k 68.8%). The 5-year survival rate of the patients who recurred and have been retreated is 70.5%. The 5-year survival rate of the patients after the appearance of a metastasis is 38.1% and 2 patients have survived more than 10 years. The NED 5-year survival rate is 48.8%. There is no significant difference in the NED 5 year survival rate according to sites or treatments. The crude 5-year survival rate is 70%, 51.4% at 10 years and 32.2% at 15 years. Our study shows that adenoid cystic carcinoma have a peculiar and slow evolution. Surgery with post operative radiotherapy obtains the best local control. These results and the radiosensibility of these lesions allow us to propose an aggressive treatment for the recurrence and the primary tumor of the directly metastatic patients. PMID- 3032875 TI - The HLA-class II-associated chondroitin sulfate proteoglycan expressed by class II positive T and monocyte-like cell lines is larger than that expressed by EBV transformed B-lymphoblastoid cell lines. AB - The human class II-associated chondroitin sulfate proteoglycan (CSPG) was originally detected as an approximately 40-70 kd species from normal human tonsil cells and Epstein-Barr virus (EBV) transformed B-lymphoblastoid cell lines. The identification of the invariant chain as the core protein of the CSPG allowed us to directly assay for the CSPG on both class II positive and negative immunocompetent cells of other lineages. Our results indicate that the CSPG is present on the class II-positive monocyte-like cell line U937 and T-cell line HUT 102, but not on the class-II negative T-cell line CCRF/CEM. No class II positive cells were found that did not also express the CSPG. The expression of the CSPG on U937 cells is increased after stimulation with gamma-interferon and PMA, paralleling the previously described increase in class II and invariant chain expression. In addition, the CSPGs from U937, HUT-102, and Raji, all cell lines derived from human malignancies, migrate as an approximately 55-90 kd species, larger than the CSPG previously characterized. However, the core proteins of the CSPG from all cells studied appear as two bands of 38 and 28 kd, indicating the size difference in the CSPG is attributable to differences in the glycosaminoglycan chains. PMID- 3032877 TI - Multi-modality treatment of primary nonresectable intrahepatic cholangiocarcinoma with 131I anti-CEA--a Radiation Therapy Oncology Group Study. AB - Thirty-seven patients with primary nonresectable intrahepatic cholangiocarcinoma (57% with prior treatment and/or metastasis) were prospectively treated with external radiation, chemotherapy, and 131I labelled anti-CEA. Therapy began in all trials with whole liver irradiation (21.0 Gy, 3.0 Gy/Fx, 4 days/week, 10 MV photons) with alternate treatment day chemotherapy (Adriamycin, 15 mg + 5-FU, 500 mg). One month after external beam therapy, chemotherapy was given (Adriamycin, 15 mg + 5-FU, 500 mg) followed the next day by the first administration of 131I anti-CEA. The treatment schedule used was 20 mCi day 0; 10 mCi day 5 as an outpatient. This schedule was derived from tumor dose estimates which indicated that 20 mCi (8-10 mCi/mg IgG) was sufficient to achieve tumor saturation with a tumor effective half-life of 3 to 5 days, depending upon the species of animal from which the antibody was obtained. The median tumor dose for the 20 mCi + 10 mCi regimen was 6.2 Gy. Antibody therapy was delivered in 2-month cycles using antibody generated in different species of animals; rabbit, pig, monkey, and bovine. Toxicity was limited to hematologic toxicity and was manifested as thrombocytopenia and leukocytopenia (3.2% Grade IV for each according to RTOG toxicity criteria). Tumor remission evaluated by CT scan digitized tumor volume analysis indicated a 26.6% partial response (PR). Tumor remission by physical examination indicated a 33.3% remission rate (25.9% PR and 7.4% complete remission (CR]. The median survival for patients who responded was 15.2 months. The actuarial median survival for the entire group of patients (metastases and previous treatment) was 6.5 months. The longest partial remission is presently more than 4 years. PMID- 3032878 TI - Timing of elective brain irradiation: a critical factor for brain metastasis-free survival in small cell lung cancer. AB - Risk factors for brain metastasis in small cell lung cancer were studied in 260 patients without brain metastasis at presentation who were treated on 5 protocols. The first three protocols offered elective brain irradiation (EBI) to all available patients after 2 or 3 courses of chemotherapy (early EBI), whereas the other two offered it after 5 or 6 courses of chemotherapy (late EBI). The overall incidence of brain metastasis was higher in the late EBI group than in the early EBI group (23.6% vs. 14.3%, p = 0.08), primarily due to higher incidence of brain metastasis developing before EBI in the former. There was a higher incidence of brain metastasis in patients without than in patients with bone marrow metastasis (21.4% vs. 6.8%, p = 0.04), in patients less than 60 years old than in patients 60 years or older (23.4% vs. 13.4%, p = 0.06), and in patients with than in patients without bone metastasis (30.2% vs. 16.8%, p = 0.07). Major risk factors for short brain metastasis-free survival were bone metastasis (p = 0.008), late EBI (p = 0.03), and failure to achieve complete remission after induction chemotherapy (p = 0.001) or as a best response (p = 0.0001). The early EBI group had a longer brain metastasis-free survival than the late EBI group, even among patients with bone metastasis (p = 0.02) or bone marrow metastasis (p = 0.05) and those who achieved complete remission after induction chemotherapy (p = 0.06) or as a best response (p = 0.05). These results indicate that timing of EBI is a critical factor in brain metastasis in patients with small cell lung cancer. There was no difference in overall survival between the two groups, however, even among patients who achieved complete remission as a best response. PMID- 3032879 TI - Immunologic and virologic findings in a bull chronically infected with noncytopathic bovine viral diarrhea virus. AB - Depressed lymphocyte blastogenesis in response to mitogen stimulation, depressed iodination of protein by neutrophils, and enhanced ingestion of Staphylococcus aureus by neutrophils were detected in a bull with chronic bovine viral diarrhea (BVD). Before developing chronic BVD, the bull was vaccinated with a killed cytopathic BVD virus. Neutralizing antibodies specific for the vaccine virus were detected in serum specimens obtained from the bull immediately before death. A noncytopathic BVD virus was isolated from the spleen after death. The immunologic and virologic findings in this bull supported reported research findings on the pathogenetic mechanisms involved in chronic BVD and mucosal disease. PMID- 3032881 TI - Effect of dietary supplements of sodium or potassium bicarbonate on short-term macromineral balance in swine. AB - Four crossbred barrows, weighing an average of 26 kg each, were fitted with simple T-cannulas in the terminal ileum and placed in metabolism cages to evaluate the effect of dietary supplements of NaHCO3 or KHCO3 on the short-term metabolism of sodium (Na), potassium (K), calcium (Ca), magnesium (Mg) and chloride (Cl). A control diet containing 2.8 g/kg Na and 4.1 g/kg K was compared with similar diets supplemented with either 13 g/kg or 26 g/kg NaHCO3 or 30 g/kg KHCO3. All diets contained 4 g/kg Cr2O3 as an external marker and were offered twice daily (1,100 g X pig-1 X d-1) in a 4 X 4 Latin square arrangement. Feces and total urine output were collected for 24 h on the fifth day after introducing a new diet; digesta was collected for 12-h periods on d 6 and 7. Sodium and K concentrations at the terminal ileum were unaffected by dietary treatment. Apparent ileal digestibility of Na was increased by NaHCO3 supplements. Over the total gastrointestinal tract, diet had no affect on apparent Na digestibility. Urinary Na clearance was increased by NaHCO3 in the diet in a dose-dependent manner. Net Na retention (g/d) was increased by NaHCO3. Apparent ileal digestibility of K was increased by KHCO3. Apparent fecal digestibility of K was increased by KHCO3 and NaHCO3. Urinary K clearance was elevated by KHCO3, but not enough to overcome the increased K intake; net K balance (g/d) rose in response to dietary KHCO3 supplements. Sodium bicarbonate or KHCO3 had no effect on short term digestibility or balance of Mg, Ca or Cl.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3032880 TI - Effect of cisplatin on hair cell morphology and lateral wall Na,K-ATPase activity. AB - The dose-response ototoxic effects of cisplatin were studied in guinea pigs. Loss of Preyer reflex and suppression of the N1 amplitude occurred in cisplatin treated animals and was described as dose-related. Drug-induced hair cell damage, as observed with scanning electron microscopy, occurred sporadically throughout the turns of the cochlea and the incidence increased with dose. Na,K-ATPase activity in the lateral wall tissues was not significantly different between treatment groups. The results reported here indicate that cisplatin ototoxicity was dose-dependent, but was not directly related to Na,K-ATPase activity in the lateral wall. PMID- 3032882 TI - Action of fluconazole (UK-49,858) in relation to other systemic antifungal azoles. AB - Viability studies showed that fluconazole (UK-49,858) was strictly fungistatic in its activity against three strains of Candida species, whether tested against yeasts in stationary or early logarithmic phase. In this regard, fluconazole appears to offer no advantage over three other oral azole-containing agents, including ketoconazole, vibunazole (Bay n 7133), and ICI 153,066. PMID- 3032883 TI - In-vitro activity of erythromycin, roxithromycin and CP 62993 against common paediatric pathogens. PMID- 3032884 TI - Comparative in-vitro activities of teicoplanin, vancomycin, coumermycin and ciprofloxacin, alone and in combination with rifampicin or LM 427, against Staphylococcus aureus. AB - The bactericidal interactions in vitro of two antibiotics active at the cell wall (teicoplanin and vancomycin) or two inhibitors of DNA gyrase (coumermycin and ciprofloxacin) combined with two inhibitors of protein synthesis (rifampicin and LM 427) were assessed against five Staphylococcus aureus strains. The S. aureus isolates included two oxacillin-sensitive and three oxacillin-resistant strains. The concentrations tested covered the range usually obtained in the serum with recommended doses. Coumermycin + rifampicin or LM 427, and ciprofloxacin + rifampicin or LM 427 were antagonistic when the concentration of either drug was higher than the MIC, and additive when sub-MIC concentrations were tested. Teicoplanin or vancomycin + rifampicin or LM 427 produced variable results. Of the synergic combinations, 14 of 15 included teicoplanin. Of the antagonistic combinations, 20 of 33 included LM 427. Antagonism with LM 427 was due to the higher intrinsic activity of LM 427 alone than teicoplanin, vancomycin or rifampicin. Teicoplanin (20 mg/l) + rifampicin or LM 427 (1 mg/l) was synergic (9/10). PMID- 3032885 TI - Cefodizime (HR 221) potentiation of human neutrophil oxygen-independent bactericidal activity. AB - The enhanced bactericidal activity of human neutrophils induced by cefotaxime and cefodizime, two methoxy-imino-amino- 2-thiazolyl cephalosporins, is linked to the cell stimulation of oxygen-dependent and oxygen-independent killing systems, respectively. Cefotaxime enhances both the killing and the oxidative response of neutrophils to opsonized particulate stimuli (bacteria for both activities and opsonized zymosan for the oxidative burst). These effects were not observed with non-opsonized particles (bacteria or zymosan) or soluble stimuli. On the contrary, cefodizime enhances killing of opsonized and non-opsonized bacteria by neutrophils regardless of treatment with phenylbutazone which blocks neutrophil oxidative metabolism. Cefodizime does not universally alter the oxidative burst induced by various stimuli, but has been shown to enhance the bactericidal activity of crude extracts of neutrophil granules. The data suggest that cefodizime and non O2-dependent killing systems of neutrophils cooperate in killing bacteria. PMID- 3032886 TI - Effects of gas composition and pH on kinetics of lung angiotensin-converting enzyme. AB - Given the pH dependence of enzymes in general and the potential importance of a blood and alveolar gas composition dependency on the interpretation of changes in the hydrolysis of angiotensin-converting enzyme (ACE) substrates by pulmonary endothelial ACE, we examined the influence of Pco2 and Po2 on the hydrolysis of a synthetic ACE substrate (benzoyl-phenylalanyl-alanyl-proline, BPAP) on passage through isolated rabbit lungs. Perfusate pH values of about 7.1, 7.4, and 7.9 were obtained by ventilating the lungs with gas containing different CO2 concentrations and Po2 values of approximately 110 and approximately 10 Torr were obtained by varying the concentration of O2 in the ventilating gas mixture. In the range studied neither acidosis nor alkalosis produced any significant changes in BPAP hydrolysis or in the kinetic parameters, Vmax and Km, for the hydrolysis process. On the other hand, a reduction in BPAP hydrolysis was detected when the Po2 was reduced from 110 to 10 Torr. The Vmax for BPAP hydrolysis by the lung was inversely correlated with the magnitude of the hypoxic vasoconstriction that occurred, suggesting that the reduced BPAP hydrolysis with hypoxia was due to the loss of perfused surface area due to the vasoconstriction. The results suggest that correlations between Pco2 and/or pH and whole-lung ACE activity that might occur in diseased lungs do not imply causalty. The hemodynamic consequences of changing Po2 (i.e., hypoxic vasoconstriction) may alter whole-organ ACE activity in the sense of changing the perfused surface area (i.e., the amount of ACE in contact with flowing perfusate). PMID- 3032887 TI - Differences in brain cytochrome responses to carbon monoxide and cyanide in vivo. AB - Cytochrome oxidation-reduction responses to two mitochondrial electron transport inhibitors, carbon monoxide (CO) and cyanide (CN), were studied in the intact brains of fluorocarbon-circulated rats. In vivo reflectance spectrophotometry indicated that cortical b-type cytochromes (564 nm) were highly resistant to reduction by CN in the presence of O2 but showed reduction responses to the administration of 1-5% CO in 90% O2. In contrast, cyanide-sensitive cytochromes aa3 (605 nm) and c + c1 (551 nm) did not increase their reduction levels during exposure to 5% CO in 90% O2. The in vivo CO-mediated b-cytochrome reduction responses did not occur after pretreatment with the cytochrome b inhibitor, antimycin A. Transmission spectrophotometry of superfused hemoglobin-free rat brain slices confirmed cortical b-type cytochromes to be CN-resistant in the presence of O2. Another cytochrome absorbing at 445 nm also was resistant to reduction by 1-mM cyanide in vitro, but it could be reduced anaerobically. The reduced 445-nm cytochrome bound CO in the presence of cyanide. We postulate that this CN-resistant CO binding component might account for in vivo cytochrome aa3 CO interactions and directly or indirectly modulate cytochrome b reduction responses to CO. In any event, the spectral data indicate different primary tissue target sites for CO and CN in living rat brain and also suggest different bioenergetic consequences of exposure to the two agents. PMID- 3032888 TI - CVF-induced decomplementation: effect on lung transvascular protein flux after thrombin. AB - We examined the effects of cobra venom factor (CVF) on the changes in pulmonary hemodynamics and transvascular fluid and protein exchange following thrombin induced pulmonary microembolism. Studies were made in unanesthetized sheep prepared with lung lymph fistulas. The animals received tranexamic acid (100 mg) to suppress fibrinolysis and were then challenged with an intravenous infusion of alpha-thrombin (80 U/kg). Control-thrombin challenged sheep were compared with the CVF-treated sheep challenged with the same thrombin dosage. CVF treatment (187 U X kg-1 X day-1 for 4 days) decreased the total hemolytic complement activity by 45% of control. Thrombin infusion in control sheep increased the mean pulmonary arterial pressure (Ppa), pulmonary vascular resistance (PVR), and lymph protein clearance (pulmonary lymph flow X lymph-to-plasma protein concentration ratio, Clym). Thrombin infusion in CVF-treated sheep produced smaller increments in Ppa, PVR, and Clym. Pulmonary lymph obtained from control-thrombin and CVF thrombin sheep induced migration of granulocytes obtained from normal unchallenged sheep. The granulocytes obtained from CVF-treated sheep responded relatively less to the migratory and O-2-generating stimuli (i.e., zymosan treated serum, pulmonary lymph from sheep after thrombin challenge, and plasma from sheep after CVF treatment) compared with normal granulocytes. The attenuation of the thrombin-induced increases in Ppa, PVR, and lung transvascular fluid and protein exchange by CVF treatment may be the result of impaired function of granulocytes. PMID- 3032889 TI - Clonal growth and culture life span of bovine adrenocortical cells in a serum free medium. AB - The life span and growth from clonal density of bovine adrenocortical cell cultures were studied in serum-supplemented medium and a serum-free defined medium, which supported sustained cell proliferation and steroid production. The total culture life span was 79 population doublings in serum-supplemented medium with fibroblast growth factor (FGF) and 36 population doublings in the defined medium without serum. Older passage cell cultures grown in the defined medium progressively lost the ability to produce 11 beta- and 21-hydroxylated steroids, which was observed previously for cultures in serum-supplemented medium, and also had a decline of 17 alpha-hydroxylated steroid production. The cloning efficiency in the defined medium was 12.2% as compared to 24% in serum-supplemented medium with FGF. Five isolated clonal cell lines grown in the defined medium were characterized for steroid function in response to steroidogenic agents. All five clonal cell lines had stimulated steroid production with 8-bromo-cAMP, but only four of the clonal lines were stimulated also by adrenocorticotropin. None of the clonal cell lines produced 11 beta-, 21- or 17 alpha-hydroxylated steroids in response to treatment with either steroidogenic agent, results that were similar to data obtained from older mass cultures. The apparent deficiency of the defined medium as compared to serum-supplemented medium for maximum support of the culture life span and cloning efficiency may be useful in studies of cellular aging and its relation to differentiated function for this cell culture system. PMID- 3032891 TI - N-terminal amino acid sequence of leukemia derived growth factor (LGF) from human erythroleukemia cell culture. AB - A human erythroleukemia cell line, K-562 T1, was adapted to a protein-free chemically defined medium; that is, the medium does not contain any proteins such as exogenous hormones, growth factors, serum and serum albumin. The K-562 T1 cells which can proliferate in a protein-free medium are one of the model systems suitably supporting the autocrine hypothesis, which claims that cancer cells produce and respond to their own growth factors. The K-562 T1 cells were cultured in a protein-free medium at large scale and the growth factors were purified from the conditioned medium. It was found that K-562 T1 cells produce at least two growth factors; one is LGF-I (leukemia-derived growth factor-I) which can stimulate the proliferation of a wide range of human leukemia cell lines and the other is LGF-II (leukemia-derived growth factor-II), which can contribute to the growth of fibroblasts. LGF-I was purified using QAE-Sephadex, Bio Gel P-60 and Mono S FPLC. The purified protein was found to be homogenous by SDS polyacrylamide gel electrophoresis and NH2-terminal sequence analysis. The molecular weight of LGF-I was 20,000 by SDS-polyacrylamide gel electrophoresis. The 30 NH2-terminal residues of LGF-I are the same as that of ubiquitin. Ubiquitin is a protein found in eukaryotic cells with molecular weight of 8,600. In the nucleus ubiquitin is conjugated to histone 2A to form the nuclear protein A24 which may play a role in regulation of chromatin structure, and in the cytoplasm is part of an ATP-dependent non-lysosomal proteolytic pathway. However, its physiological significance has not yet been fully resolved. Ubiquitin purified from bovine thymus did not show cell proliferating activity for any cells tested. The results suggest that LGF-I is a new autocrine growth factor with a molecular weight of 20,000 daltons, containing ubiquitin at the NH2 terminal end. PMID- 3032892 TI - General Referee reports: Committee on Feeds, Fertilizers, and Related Materials. PMID- 3032890 TI - Collagen-fibronectin interactions in normal and Rous sarcoma virus-transformed avian tendon cells: possible mechanisms for increased extracellular matrix turnover after transformation. AB - Using gelatin, casein, and fibronectin as substrates and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), we have identified protein degrading enzymes in both normal and Rous sarcoma virus-transformed primary avian tendon cells. Although there are some consistent differences in the profile of the gelatinolytic activities (mainly metalloproteinases) between normal and transformed cells, the amounts of fibronectin-degrading activities seem to be comparable. In vitro studies reported here demonstrate that the degradation of fibronectin is partially and specifically inhibited by gelatin and collagen. We therefore propose that the abundant collagen present in normal tendon cells protects fibronectin against degradation. Conversely, in transformed cells, where collagen levels are drastically reduced, fibronectin may be more accessible to degradation. Thus differences in the steady-state levels of fibronectin on normal and transformed cells may be, at least in part, a consequence of changes in collagen levels. PMID- 3032893 TI - [Echographic and computerized tomography aspects of pseudomyxoma peritonei: apropos of a case and review of the literature]. PMID- 3032894 TI - Genetic analysis in Salmonella typhimurium with a small collection of randomly spaced insertions of transposon Tn10 delta 16 delta 17. AB - We report the isolation of a group of 279 Salmonella typhimurium strains carrying randomly spaced insertions of the minitransposon Tn10 delta 16 delta 17 and describe the use of these strains to facilitate genetic analysis. The insertions were isolated initially in individual recombinant lambda clones from a genomic library. Individual insertions were then moved into the S. typhimurium chromosome, where the distribution of insertion sites relative to standard genetic markers was analyzed in a series of transductional crosses. Since a different, randomly chosen clone was used to generate each insertion, the distribution of insertion positions should have been as random as the cloning events leading to the formation of the library. In agreement with this expectation, most S. typhimurium markers tested were cotransducible with one or more of these Tn10 delta 16 delta 17 insertions. We expect that most new mutations will be quickly classified and mapped by determination of the pattern of cotransduction with this set of insertions. This use is illustrated by the analysis of a group of lac operon fusions regulated by anaerobiosis. We also describe several other applications that should make this collection a useful new tool in S. typhimurium genetics. PMID- 3032895 TI - Mutations in the lac P2 promoter. AB - We used site-directed mutagenesis to generate mutations in the -10 region of the lac P2 promoter. The mutations were crossed onto lambda bacteriophage carrying the lac regulatory elements and an intact lacZ gene, and the effects of the various mutations were determined in vivo and in vitro. Two of four mutations had effects on the start point of the P2-directed transcript and had very little effect on lac expression. Another mutation, which abolishes P2 promoter activity in vitro, also had very little effect on lac expression in vivo. We suggest that the P2 promoter plays little or no role in the activation of the P1 promoter by catabolite activator protein in complex with cyclic AMP. PMID- 3032896 TI - Molecular cloning of a recA-like gene from the cyanobacterium Anabaena variabilis. AB - A recA-like gene isolated from the cyanobacterium Anabaena variabilis was cloned and partially characterized. When introduced into Escherichia coli recA mutants, the 7.5-kilobase-pair plasmid-borne DNA insert restored resistance to methyl methanesulfonate and UV irradiation, as well as recombination proficiency when measured by Hfr-mediated conjugation. The cyanobacterial recA gene restored spontaneous but not mitomycin C-induced prophage production. Restriction analysis and subcloning yielded a 1.5-kilobase-pair Sau3A fragment which also restored methylmethane sulfonate resistance and coded for a 38- to 40-kilodalton polypeptide when expressed in an in vitro transcription-translation system. PMID- 3032898 TI - Two families of repeated DNA sequences in Thiobacillus ferrooxidans. AB - The genome of Thiobacillus ferrooxidans ATCC 19859 is about 2.8 X 10(6) base pairs as determined by analysis of reassociation kinetics of sheared DNA. This is 70% of the size of the genome of Escherichia coli. About 6% of the genome of T. ferrooxidans consists of moderately repetitive DNA sequences that are repeated an average of 20 times per genome. Two distinct repeated sequences, designated family 1 and family 2, have been analyzed in more detail. Both families are approximately 1 kilobase in length and are repeated 20 to 30 times per genome. Preliminary evidence from restriction enzyme analysis, Southern blotting experiments, and thermal melting analysis indicates that members of both families are conserved and are interspersed with single-copy DNA. Six copies of one family are present on the 45-kilobase-pair plasmid of strain ATCC 19859. PMID- 3032897 TI - Nucleotide sequence analysis of RepFIC, a basic replicon present in IncFI plasmids P307 and F, and its relation to the RepA replicon of IncFII plasmids. AB - RepFIC is a basic replicon of IncFI plasmid P307 which is located within a 3.09 kilobase SmaI fragment. The nucleotide sequence of this region has been determined and shown to be homologous with the RepFIIA replicon of IncFII plasmids. The two replicons share three homologous regions, HRI, HRII, and HRIII, which are flanked by two nonhomologous regions, NHRI and NHRII. A comparison of coding regions reveals that the two replicons have several features in common. RepFIC, like RepFIIA, codes for a repA2 protein with its amino-terminal codons in HRI and its carboxy-terminal codons in NHRI. Although the codons for the repA1 proteins are located in NHRII, the DNA region containing a putative promoter, ribosomal binding site, and initiation codons is located in HRII. This region also codes for an inc RNA. There are nine base-pair differences between the inc RNA of RepFIIA and that of RepFIC, and as a result, RepFIC and RepFIIA replicons are compatible. An EcoRI fragment from the F plasmid which shows homology with RepFIC of P307 has also been sequenced. This fragment contains only a portion of RepFIC, including the genes for the putative repA2 protein and inc RNA. The region coding for a putative repA1 protein is interrupted by the transposon Tn1000 and shows no homology with the repA1 region of RepFIIA and RepFIC of P307. Our comparative and structural analyses suggest that RepFIC and RepFIIA, although different, have a similar replication mechanism and thus can be assigned to the same replicon family, which we designate the RepFIIA family. PMID- 3032899 TI - Overexpression of the dnaA gene in Escherichia coli B/r: chromosome and minichromosome replication in the presence of rifampin. AB - The replication of chromosomes and minichromosomes in Escherichia coli B/r was examined under conditions in which the dnaA gene product was overproduced. Increased levels of the DnaA protein were achieved by thermoinduction of the dnaA gene, under the control of the lambda pL promoter, or by cellular maintenance of multicopy plasmids carrying the dnaA gene under the control of its own promoters. Previous work has shown that overproduction of DnaA protein stimulates replication of the chromosomal origin, oriC, but that the newly initiated forks do not progress along the length of the chromosome (T. Atlung, K. V. Rasmussen, E. Clausen, and F. G. Hansen, p. 282-297, in M. Schaechter, F. C. Neidhardt, J. L. Ingraham, and N. O. Kjeldgaard, ed., The Molecular Biology of Bacterial Growth, 1985). In the present study, it was found that overproduction of DnaA protein caused both a two- to threefold increase in the amount of residual chromosome replication and an extended synthesis of minichromosome DNA in the presence of rifampin. The amount of residual chromosome replication was consistent with the appearance of functional replication forks on the majority of the chromosomes. Since the rate of DNA accumulation and the cellular DNA/mass ratios were not increased significantly by overexpression of the dnaA gene, we concluded that the addition of rifampin either enabled stalled replication forks to proceed beyond oriC or enabled new forks to initiate on both chromosomes and minichromosomes, or both. PMID- 3032900 TI - Mobilization of Haemophilus influenzae chromosomal markers by an Escherichia coli F' factor. AB - Filter matings between E. coli K-12 strains carrying an F'::Tn5,Tn9 factor with H. influenzae Rd strains gave rise to kanamycin-chloramphenicol-resistant H. influenzae strains at a frequency of approximately 10(-6). Transfer of the F' factor to H. influenzae was verified by expression of unselected markers in H. influenzae (lac+ or cotransfer of the nonselected antibiotic resistance), physical presence of a high-molecular-weight plasmid in recipient H. influenzae cells, and detection by Southern hybridization analysis of DNA sequences specific for the F' factor replication and partition functions in recipient H. influenzae cells. H. influenzae (F' Tn5,Tn9) strains were capable of transferring kanamycin and chloramphenicol resistances to other H. influenzae strains and were capable of mobilizing H. influenzae chromosomal markers at a low frequency. Insertion of a Tn5 element in the H. influenzae genome near the novobiocin resistance gene increased the frequency of transfer of novobiocin resistance about 30-fold. Transfer of other chromosomal markers also increased, although to a lesser extent, and ordered transfer of chromosomal markers could be demonstrated. Gene transfer was insensitive to DNase I, and transfer of chromosomal (but not F' factor) markers was dependent on the H. influenzae rec-1 and rec-2 gene functions. PMID- 3032901 TI - Inactivation of the ampD gene causes semiconstitutive overproduction of the inducible Citrobacter freundii beta-lactamase. AB - In Citrobacter freundii and Enterobacter cloacae, synthesis of AmpC beta lactamase is inducible by the addition of beta-lactams to the growth medium. Spontaneous mutants that constitutively overproduce the enzyme occur at a high frequency. When the C. freundii ampC beta-lactamase gene is cloned into Escherichia coli together with the regulatory gene ampR, beta-lactamase expression from the clone is inducible. Spontaneous cefotaxime-resistant mutants were selected from an E. coli strain carrying the cloned C. freundii ampC and ampR genes on a plasmid. Virtually all isolates had chromosomal mutations leading to semiconstitutive overproduction of beta-lactamase. The mutation ampD2 in one such mutant was caused by an IS1 insertion into the hitherto unknown ampD gene, located between nadC and aroP at minute 2.4 on the E. coli chromosome. The wild type ampD allele cloned on a plasmid could fully trans-complement beta-lactamase overproducing mutants of both E. coli and C. freundii, restoring the wild-type phenotype of highly inducible enzyme synthesis. This indicates that these E. coli and C. freundii mutants have their lesions in ampD. We hypothesize that induction of beta-lactamase synthesis is caused by blocking of the AmpD function by the beta-lactam inducer and that this leads directly or indirectly to an AmpR mediated stimulation of ampC expression. PMID- 3032902 TI - Regulation of cell division in Escherichia coli K-12: probable interactions among proteins FtsQ, FtsA, and FtsZ. AB - In Escherichia coli, the FtsQ, FtsA, and FtsZ proteins are believed to play essential roles in the regulation of cell division. Of the three proteins, FtsZ has received the most attention, particularly because of its interactions with SfiA. Double mutants which carry mutations located in the ftsQ, ftsA, or ftsZ gene in combination with the lon-1 mutation were constructed. In the presence of the lon-1 mutation, which is known to stabilize SfiA, the ftsQ1 mutant cells were not capable of forming colonies on a rich agar medium, whereas mutant cells harboring either one of the mutations grew well on this medium. Examination of lon-1 fts double-mutant cells for sensitivity to UV light revealed that those carrying the ftsA10 allele were resistant. It was also observed that in the presence of a multicopy plasmid containing a wild-type ftsZ gene, the ftsQ1 mutant filamented markedly following a nutritional shift-up and that the division rate of ftsZ84 mutant cells was slightly reduced when they harbored a wild-type ftsQ-containing plasmid. The possibility that the Fts proteins are interacting with one another and forming a molecular complex is discussed. PMID- 3032903 TI - Defective regulation of the phenylalanine biosynthetic operon in mutants of the phenylalanyl-tRNA synthetase operon. AB - Among mutants of Escherichia coli resistant to p-fluorophenylalanine (PFP) were some with constitutive expression of the phenylalanine biosynthetic operon (the pheA operon). This operon is repressed in the wild type by phenylalanine. The mutation in three of these mutants mapped in the aroH-aroD region of the E. coli chromosome at 37 min. A plasmid bearing wild-type DNA from this region restored p fluorophenylalanine sensitivity and wild-type repression of the pheA operon. Analysis of subclones of this plasmid and comparison of its restriction map with published maps indicated that the mutations affecting regulation of the pheA operon lie in the structural genes for phenylalanyl-tRNA synthetase, pheST, probably in pheS. Thus, the pheST operon has a role in the regulation of phenylalanine biosynthesis, the most likely being that wild-type phenylalanyl tRNA synthetase maintains a sufficient intracellular concentration of Phe tRNA(Phe) for attenuation of the pheA operon in the presence of phenylalanine. A revised gene order for the 37-min region of the chromosome is reported. Read clockwise, the order is aroD, aroH, pheT, and pheS. PMID- 3032904 TI - Competence for genetic transformation in Streptococcus pneumoniae: molecular cloning of com, a competence control locus. AB - To identify and map genes involved in competence for genetic transformation in Streptococcus pneumoniae, we have cloned DNA surrounding an ermB insertion mutation that causes a competence factor deficiency. We recovered the insert and approximately 500 base pairs of neighboring pneumococcal DNA in pMB9. Larger pieces of DNA from this region were unstable in pMB9 and pBR325. However, larger pieces were stable in pKK232-8, an Escherichia coli vector containing strong transcription terminators. Overlapping pieces of wild-type DNA from this competence control region were cloned and mapped in this vector. Insertion mutations were constructed in vitro throughout the cloned region. When crossed into the pneumococcus chromosome, they showed that the com locus was 4.2 to 5.2 kilobases long. PMID- 3032905 TI - Molecular cloning of Mu d(bla lacZ) transcriptional and translational fusions. AB - The vector pBW2 was made to selectively clone chimeric plasmids with chromosomal Mu d(bla lacZ) transcriptional or translational fusions. It was tetracycline resistant and had the carboxyl-terminal end of bla distal to its PstI site. Because ligation of PstI-digested chromosomal DNA of a Mu d(bla lacZ) insertion with pBW2 restored bla, ampicillin-resistant chimeric plasmids were selectable. These plasmids had the Mu d bla amino terminus and simultaneously acquired other Mu d sequences including lacZ, the chromosomal fusion joint, and the DNA adjacent to the nearest chromosomal PstI site. The plasmid pBW2 was useful in the molecular cloning of several psi and pho::lacZ(Mu d) fusions, as well as chromosomal genes located near Mu d insertions. PMID- 3032906 TI - Ferric-coprogen receptor FhuE of Escherichia coli: processing and sequence common to all TonB-dependent outer membrane receptor proteins. AB - Iron transport via siderophores requires outer membrane receptor proteins and the TonB protein. The FhuE protein of Escherichia coli functions as the receptor for ferric coprogen and ferric-rhodotorulic acid. A chromosomal DNA fragment bearing the fhuE gene was cloned into pACYC184. The gene was localized by insertion mutagenesis by using the transposon Tn1000. Expression in minicells revealed a FhuE precursor with an apparent molecular weight of 82,000 and a FhuE protein with a molecular weight of 76,000. The transcription polarity of the fhuE gene was deduced from the size of truncated polypeptides derived from Tn1000 insertions, which were mapped by restriction analysis. The processing of truncated precursors that were synthesized by insertion mutants was strongly reduced even when the insertion site was close to the carboxy terminus of the FhuE protein. It is concluded that either the efficient insertion of proFhuE into the cytoplasmic membrane or the rate of cleavage of the signal peptide requires a particular conformation of the proFhuE protein, which is only formed by the complete primary structure. The amino-terminal amino acid sequence deduced from the nucleotide sequence was confirmed by gas-phase sequencing of the precursor and the mature form, which were separated by electrophoresis on polyacrylamide gels. The precursor contained an unusually long signal peptide of 36 amino acids. The amino-terminal end of the mature form contained the sequence Glu-Thr-Val Ile Val. A pentapeptide starting with either Glu or Asp, followed by Thr, and two uncharged residues ending with Val were found in all outer membrane receptor proteins that were constituents of TonB-dependent transport systems. PMID- 3032907 TI - Identification of the cydC locus required for expression of the functional form of the cytochrome d terminal oxidase complex in Escherichia coli. AB - The aerobic respiratory chain of Escherichia coli contains two terminal oxidases which are differentially regulated. The cytochrome o complex predominates under growth conditions of high aeration, whereas the cytochrome d complex predominates when the oxygen tension is low. Either terminal oxidase will support aerobic growth. The goal of the work presented in this paper was to identify genes required for the expression of the functional form of the cytochrome d complex, other than the genes encoding the polypeptide components of the oxidase complex (cyd locus). A strain lacking the cytochrome o complex (cyo mutant strain) was mutagenized by using a lambda-Mu hybrid hopper bacteriophage, lambda placMu53, which inserts randomly into the chromosome and carries a kanamycin resistance marker. Strains were isolated and examined which were unable to grow aerobically, i.e., which lacked functional cytochrome d complex, and which could not be complemented by introduction of the cyd gene on F-prime episomes. One strain was selected for characterization. The phage insert was mapped to min 18.9 on the genetic linkage map, defining a new genetic locus, cydC. Evidence described in the text suggests that the gene product is probably required for the synthesis of the unique heme d component of the cytochrome d complex. PMID- 3032909 TI - Regulation in Escherichia coli of the porin protein gene encoded by lambdoid bacteriophages. AB - Specialized lambda transducing phages carrying the cloned lc porin gene from the lambdoid bacteriophage PA-2, including various amounts of a sequence 5' to the start of transcription, were used to study the regulation of the porin gene. It was found that a cyclic AMP receptor protein consensus binding site 65 base pairs 5' to the start of transcription was required for catabolite repression of lc but was not sufficient for maximum expression under derepressing conditions. A sequence located more than 209 base pairs 5' to the start of transcription was necessary for maximum expression. By manipulating the copy number of the lc gene and the temperature and by measuring both the rate of synthesis of mRNA and the amount of Lc protein in the outer membrane, it was determined that the expression of lc is regulated primarily at the level of transcription and that expression is not autoregulated. Evidence is also presented that the silent phage porin gene nmpC of Escherichia coli K-12 is transcribed to the same extent as lc even though it does not give rise to a stable pool of mRNA. The structure of the 5' end of lc and nmpC is similar to that of ompF, and a model for transcriptional regulation is presented which may apply to all of these porin genes. PMID- 3032908 TI - Transfer of the conjugal tetracycline resistance transposon Tn916 from Streptococcus faecalis to Staphylococcus aureus and identification of some insertion sites in the staphylococcal chromosome. AB - The Streptococcus faecalis pheromone-dependent conjugative plasmid pAD1::Tn916 and the membrane filter-dependent conjugative plasmid pPD5::Tn916 were used to introduce Tn916 into Staphylococcus aureus by intergeneric protoplast fusions and intergeneric membrane-filter matings. In recombinants obtained by protoplast fusion where no plasmid DNA could be detected, tetracycline resistance resulted from transposition of Tn916 from pAD1 to the S. aureus chromosome. Transformation analyses showed that S. aureus Tn916 chromosomal insertions occurred near pig, ilv, uraA, tyrB, fus, ala, and the trp operon. DNA hybridization analyses of EcoRI- and HindIII-digested chromosomal DNAs confirmed the diversity of chromosomal sites involved and demonstrated that the inserts were Tn916 insertions rather than integrations of all or part of pAD1::Tn916. Both pAD1::Tn916 and pPD5::Tn916 were transferred to S. aureus by membrane-filter matings. These plasmids remained intact and expressed tetracycline resistance in S. aureus. S. aureus strains carrying pAD1::Tn916, but not a chromosomal insert of Tn916, and any one of several conjugal gentamicin-resistance plasmids lost their ability to serve as conjugal donors of the gentamicin-resistance plasmids. PMID- 3032910 TI - Isolation and characterization of symbiotic mutants of bradyrhizobium sp. (Arachis) strain NC92: mutants with host-specific defects in nodulation and nitrogen fixation. AB - Random transposon Tn5 mutagenesis of Bradyrhizobium sp. (Arachis) strain NC92, a member of the cowpea cross-inoculation group, was carried out, and kanamycin resistant transconjugants were tested for their symbiotic phenotype on three host plants: groundnut, siratro, and pigeonpea. Two nodulation (Nod- phenotype) mutants were isolated. One is unable to nodulate all three hosts and appears to contain an insertion in one of the common nodulation genes (nodABCD); the other is a host-specific nodulation mutant that fails to nodulate pigeonpea, elicits uninvaded nodules on siratro, and elicits normal, nitrogen-fixing nodules on groundnut. In addition, nine mutants defective in nitrogen fixation (Fix- phenotype) were isolated. Three fail to supply symbiotically fixed nitrogen to all three host plants. Surprisingly, nodules elicited by one of these mutants exhibit high levels of acetylene reduction activity, demonstrating the presence of the enzyme nitrogenase. Three more mutants have partially effective phenotypes (Fix +/-) in symbiosis with all three host plants. The remaining three mutants fail to supply fixed nitrogen to one of the host plants tested while remaining partially or fully effective on the other two hosts; two of these mutants are Fix in pigeonpea and Fix +/- on groundnut and on siratro, whereas the other one is Fix- on groundnut but Fix+ on siratro and on pigeonpea. These latter mutants also retain significant nodule acetylene reduction activity, even in the ineffective symbioses. Such bacterial host-specific fixation (Hsf) mutants have not previously been reported. PMID- 3032912 TI - Identification of the promoter for a peptide antibiotic biosynthesis gene from Bacillus brevis and its regulation in Bacillus subtilis. AB - Tyrocidine is a cyclic decapeptide antibiotic which is produced and secreted by stationary-phase cells of the sporeforming bacterium Bacillus brevis. We identified the promoter for the B. brevis structural gene (tycA) for tyrocidine synthetase I, the enzyme catalyzing the first step in tyrocidine biosynthesis, and studied its regulation in cells of B. brevis and Bacillus subtilis. Transcription from the tycA promoter was induced at the end of the exponential phase of the growth cycle in B. brevis cells growing in sporulation medium. To study the regulation of tycA in B. subtilis, we constructed a derivative of the B. subtilis bacteriophage SP beta containing a transcriptional fusion of the tycA promoter to the lacZ gene of Escherichia coli and introduced the tycA-lacZ operon fusion by means of specialized transduction into sporulation mutants known to be blocked in sporulation-associated antibiotic production. Our principal finding was that tycA-directed lacZ expression was impaired in the stage-0 mutants with mutations spo0A, spo0B, and spo0E but not in spo0C, spo0F, spo0H, or spo+ bacteria. The dependence on the spo0A gene product could be entirely bypassed by an abrB suppressor mutation, which caused tycA-lacZ to be transcribed constitutively at all stages of growth. A simple model is proposed for the mechanism of tycA induction based on the Spo0A-dependent inactivation of Ab-B protein, which is proposed to be a negative regulator of tycA transcription. PMID- 3032911 TI - Outer membrane protein mediating iron uptake via pyoverdinpss, the fluorescent siderophore produced by Pseudomonas syringae pv. syringae. AB - In an iron-limited environment Pseudomonas syringae pv. syringae B301D produces a yellow-green fluorescent siderophore called pyoverdinpss which functions in high affinity iron transport. Two-dimensional electrophoretic comparisons of the outer membrane proteins of strain B301D identified nine proteins which were expressed at low (50 nM) but not at high (10 microM) iron concentrations. Except for the minor protein 8e, the iron-regulated proteins exhibited high molecular weights ranging from approximately 74,000 to 80,000. A mutant of strain B301D incapable of iron uptake (Iu-) from ferric pyoverdinpss lacked the 74,000-molecular-weight protein 4a, which was the major iron-regulated outer membrane protein. In contrast, a nonfluorescent mutant (Flu-) unable to synthesize pyoverdinpss showed no quantitative or qualitative difference in its outer membrane profile from that of the wild-type strain. In plant pathogenicity tests the Iu- and Flu- strains caused typical brown necrotic and sunken lesions in immature sweet cherry fruit which were indistinguishable from those of the wild-type strain. Thus, excretion of pyoverdinpss and subsequent Fe(III) uptake do not have a determinative role in the pathogenicity or virulence of P. syringae pv. syringae. PMID- 3032913 TI - Regulation of cobalamin biosynthetic operons in Salmonella typhimurium. AB - Transcription of cobalamin (cob) biosynthetic genes in Salmonella typhimurium is repressed by cobalamin and by molecular oxygen. These genes seem to be subject to catabolite repression, and they are maximally expressed under conditions of anaerobic respiration of glycerol-fumarate. A 215-fold increase in the expression of cob genes occurs when S. typhimurium shifts from aerobic growth on glucose to anaerobic respiration of glycerol-fumarate under strictly anoxic growth conditions. Exogenous cyclic AMP substantially stimulates the transcription of cob-lac fusions during aerobic growth. However, cyclic AMP is not absolutely required for the expression of the pathway, nor does it mediate the aerobic control. Cobalamin biosynthesis is not seen under aerobic growth conditions, even when transcription is stimulated by the addition of cyclic AMP. Hence, additional control mechanisms triggered by the presence of molecular oxygen must operate independently from transcription effects on the cob operons. PMID- 3032914 TI - Cyclic AMP phosphodiesterase in Thermomonospora curvata. AB - Cyclic AMP phosphodiesterase (PDE; EC 3.1.4.17) in Thermomonospora curvata was purified and characterized. Fractionation of cell extracts by ion-exchange and size-exclusion chromatography revealed four PDE isozymes, which differed markedly in molecular weight, theophylline sensitivity, pH optima, and substrate affinity. Although the enzyme was labile after purification, total recovery of PDE activity was fivefold that of the crude extract. PDE biosynthesis appeared sensitive to the growth phase, growth rate, and carbon source. PDE levels in batch cultures peaked and declined rapidly during mid-exponential-phase growth. In continuous culture, maximal PDE and cellulase production occurred at dilution rates yielding mean cell generation times of about 5 and 17 h, respectively. The addition of glucose to cellulose-grown cells caused declines in both cyclic AMP and PDE levels, suggesting that the enzyme was subject to, rather than the agent of, catabolite repression. PMID- 3032916 TI - Characterization by EPR spectroscopy of cytochrome b-562 isolated from the cytochrome b-c1 complex of Rhodopseudomonas sphaeroides R-26. AB - The EPR spectra of cytochrome b-562 isolated from the cytochrome b-c1 complex of Rhodopseudomonas sphaeroides were measured at liquid helium temperature. The purified cytochrome b-562 gives a high spin signal at g = 6.0. Anaerobic titration of this signal confirmed the presence of two redox components with Em = 40 and -110 mV at pH 7.5. These values are consistent with the published ones, Em = 55 and -100 mV at pH 7.0, that were optically estimated for the same type of preparation (Iba et al. (1985) FEBS Lett. 183, 151-154). The power saturation behavior of the g = 6.0 signal at different redox potentials indicated a direct spin-spin interaction between these two redox centers. PMID- 3032915 TI - Some properties of subunits of DNA gyrase from Pseudomonas aeruginosa PAO1 and its nalidixic acid-resistant mutant. AB - Subunits A and B of DNA gyrase were purified from Pseudomonas aeruginosa PAO1 and its mutant, which was resistant to nalidixic acid. Inhibition tests of DNA gyrases reconstituted with a combination of subunits from the two strains showed that an alteration of subunit A but not subunit B caused bacteria to resist fluoroquinolones. PMID- 3032917 TI - 2,6-Dichlorophenolindophenol-reducing activity of phagocytosis-associated NADPH oxidase. AB - The 2,6-dichlorophenolindophenol (DCIP)-reducing activity of the phagocytosis associated NADPH oxidase was investigated using homogenates and a membrane fraction (F2) of elicited guinea pig peritoneal macrophages stimulated by phorbol myristate acetate. Essentially all of the stimulation-specific DCIP reduction under aerobic conditions could be inhibited when high concentrations of superoxide dismutase (SOD), about 10 times those usually used to inhibit the superoxide (O-2)-mediated cytochrome c reduction, were used. SOD inhibited the DCIP reduction by chemically generated O2- in the same manner as the stimulation specific DCIP reduction by the macrophage F2, and the concentration of SOD necessary for 50% inhibition was about 10 times that for the reduction of cytochrome c. Under anaerobic conditions, however, the NADPH oxidase could reduce DCIP, though the rate was slow because we could not use a sufficiently high DCIP concentration. The observations indicate that the NADPH oxidase preferentially reduces oxygen under aerobic conditions, though the oxidase can reduce DCIP in the anaerobic state. PMID- 3032919 TI - Endogenous inhibitor of [3H]kainate binding to synaptic membrane in rat brain. AB - An understanding of the mechanism of kainic acid toxicity to neurons could provide important clues to pathogenesis of Huntington's chorea. The existence of high-affinity binding sites for kainate, a foreign compound, is suggestive of the existence of kainate-like substances in the brain. In addition to such neurotoxic kainate-like substances, and endogenous inhibitor of kainate binding may also exist in the brain to allow the synaptic function to operate normally. Based on this idea, the existence of molecules which inhibit [3H]kainate binding to synaptic membranes was examined in rat brain. An endogenous inhibitor of [3H]kainate binding to synaptic membranes was found in the supernatant obtained from synaptic membranes of rat brain. The inhibitor is a thermostable, basic protein with a relatively low molecular weight. PMID- 3032918 TI - Partial purification and characterization of exudate gelatinase in the acute phase of carrageenin-induced inflammation in rats. AB - Gelatinase has been partially purified from exudate in the acute phase of carrageenin-induced inflammation in rats. The enzyme occurs in a latent form that can be activated with 4-aminophenylmercuric acetate (APMA). The latent gelatinase was separated into an active gelatinase and a protein fraction by zinc-chelating Sepharose 6B column chromatography in the final step of purification, suggesting that the latent gelatinase is an enzyme-inhibitor complex. The pH optimum of the active gelatinase is about 7.5 and no reactivity toward native type I collagen or alpha-casein was detected. The molecular weights of the latent and active gelatinases were about 245,000 and about 185,000, respectively, as determined by gel filtration on Sephadex G-200. On the other hand, both latent and active gelatinases occurred in multiple forms in SDS-substrate polyacrylamide gel electrophoresis; the latent gelatinase showed two bands with molecular weights of 105,000 and 69,000, and two additional bands of 88,000 and 83,000 appeared when the latent gelatinase was activated with APMA, while the active gelatinase showed all four species. The active gelatinase was inhibited by metallo-proteinase inhibitors, but not by serine- or cysteine-proteinase inhibitors, suggesting that the exudate gelatinase is a metallo-proteinase. The active gelatinase was also inhibited by serum proteins such as albumin and gamma-globulin, suggesting that gelatinase does not remain in an active form in the inflammatory lesion, where the vascular permeability is increased. PMID- 3032920 TI - Epidermal growth factor increases c-myc mRNA without eliciting phosphoinositide turnover, protein kinase C activation, or calcium ion mobilization in Swiss 3T3 fibroblasts. AB - Incubation of quiescent cultures of Swiss 3T3 cells with epidermal growth factor (EGF) caused an increase in c-myc mRNA. Under these conditions, EGF did not induce phosphoinositide turnover, formation of diacylglycerol, formation of inositol tris-, bis-, and monophosphates, protein kinase C activation, or Ca2+ mobilization. Although it has been reported that both protein kinase C and Ca2+ may be responsible for the platelet-derived growth factor- and fibroblast growth factor-induced increases in c-myc mRNA in Swiss 3T3 cells (Kaibuchi, K., Tsuda, T., Kikuchi, A., Tanimoto, T., Yamashita, T., & Takai, Y. (1986) J. Biol. Chem. 261, 1187-1192), these results indicate that neither protein kinase C nor Ca2+ is involved in the EGF-induced increase in c-myc mRNA, and that an unidentified system may be involved in this reaction. PMID- 3032921 TI - Structure of the trypsin-binding domain of Bowman-Birk type protease inhibitor and its interaction with trypsin. AB - The crystal structure of the complex formed by bovine trypsin and Bowman-Birk type protease inhibitor AB-I extracted from azuki beans (Vigna angularis) 'Takara' has been analyzed. The structure was solved by the application of the phase combination of single isomorphous phases and trypsin model phases, followed by phase improvement using the iterative Fourier technique. From the resulting electron density map, a three-dimensional atomic model of the trypsin binding domain of AB-I has been built. The peptide chain at the trypsin reactive site turns back sharply at Pro29 and forms a 9-residue ring (Cys24-Cys32). The 'front side' of this ring, consisting of the reactive site (Cys24-Met28), interacts with trypsin in a similar manner to other families of inhibitors and forms a stable complex, which seems to be maintained by the interactions with the 'back side' of this ring (Pro29-Cys34). The similar spatial arrangements of the 'back side' of this inhibitor and the 'secondary contact region' of the other inhibitors with respect to the reactive site suggest an important common role of these regions in exhibiting inhibitory activity. PMID- 3032922 TI - Analysis of smooth muscle myosin phosphorylation with native pyrophosphate gels and its application to studies on myosin phosphorylation in contracting smooth muscle. AB - Gizzard smooth muscle myosin, the 20,000 Mr light chain (L20) of which had been phosphorylated in vitro with a calmodulin-myosin light chain kinase system, was separated into 5 isolated bands in a pyrophosphate polyacrylamide gel. Their mobilities were in the following order: myosin with 2 unphosphorylated L20 (GM) less than myosin with 1 unphosphorylated and 1 mono-phosphorylated L20 (GMP1) less than myosin with 2 mono-phosphorylated L20 (GMP2) less than myosin with 1 mono-phosphorylated and 1 di-phosphorylated L20 (GMP3) less than myosin with 2 di phosphorylated L20 (GMP4). We used this pyrophosphate polyacrylamide gel electrophoresis to analyze the phosphorylated state of taenia coli smooth muscle during K+-induced contraction. During the initial 2 min contraction, phosphorylated forms corresponding to GMP1 and GMP2 were detected in addition to the unphosphorylated form. PMID- 3032923 TI - Purification and properties of dihydrogeodin oxidase from Aspergillus terreus. AB - The last step of (+)-geodin biosynthesis is a phenol oxidative coupling, which is one of the most important reactions in biosynthesis of natural products. The enzyme named dihydrogeodin oxidase catalyzes the regio- and stereospecific phenol oxidative coupling reaction to form (+)-geodin from dihydrogeodin. The enzyme was purified from the cell-free extract of Aspergillus terreus, a (+)-geodin producer, by ammonium sulfate fractionation, acid treatment, and column chromatographies on DEAE-cellulose, Hydroxyapatite, chromatofocusing, and Toyopearl HW-55S. The purified enzyme was homogeneous as judged by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The molecular weight of the enzyme was estimated to be 153,000 by gel filtration on a Toyopearl HW-55S column and 76,000 by SDS-polyacrylamide gel electrophoresis, indicating that the enzyme is a dimer. The purified enzyme showed an intense blue color and had absorption maxima at 280 and 600 nm, which suggested it to be a blue copper protein. The copper content was found to be 8 atoms per subunit by atomic absorption analysis and no significant amount of other metals was detected by ICP emission spectrometry. The electron paramagnetic resonance spectrum showed the presence of type 1 and type 2 copper atoms in the enzyme molecule. Sodium azide and ethylxanthate inhibited the enzyme activity, but potassium cyanide and diethyldithiocarbamate, both known as potent copper enzyme inhibitors, were not inhibitory. PMID- 3032924 TI - Primary structure of rat liver mannan-binding protein deduced from its cDNA sequence. AB - cDNA clones encoding rat liver mannan-binding protein (MBP), a lectin specific for mannose and N-acetylglucosamine, were isolated from a rat liver cDNA library carried in lambda gt 11, by screening with affinity purified polyclonal rabbit anti-rat liver MBP antibodies. The nucleotide sequence of the cDNA determined by the dideoxy method revealed the complete amino acid sequence of the MBP (226 residues). The NH2-terminal residue of the MBP, glutamic acid, was preceded by a predominantly hydrophobic stretch of 18 amino acids, which was assumed to be a signal peptide. Near the NH2-terminal, there was a collagen-like domain, which consisted of 19 repeats of the sequence Gly-X-Y. Here, X and Y were frequently proline and lysine. Three proline and lysine residues were hydroxylated, and one of the latter appeared to link to galactose. Computer analysis of several lectins for sequence homology suggested that the COOH-terminal quarter of the MBP is associated with the calcium binding as well as carbohydrate recognition. PMID- 3032925 TI - Flash-induced proton release in Rhodopseudomonas sphaeroides spheroplasts. AB - Proton release by flash excitations was measured with right-side-out vesicles prepared from Rhodopseudomonas sphaeroides by lysozyme-EDTA treatment followed by hypotonic treatment. Absorbance change at 586 nm in the presence of bromcresol purple was measured to monitor the pH change. In the presence of horse heart cytochrome c, which catalyzes the electron transfer from the cytochrome b-c1 complex to the primary electron donor, the single-turnover flash elicited release of about two protons per primary electron donor, which was rereduced rapidly by the added cytochrome c. The halftime of the proton release was about 70 ms at pH 6.3 and at a redox potential of about 150 mV. The rate was considerably lower than that of the electron transfer from the cytochrome b-c1 complex to cytochrome c. However, multiple flashes with intervals of 60 ms caused release of the same amount of protons as that by flashes with longer intervals. This indicated that the proton release itself was rapid, but delocalization was slower. Antimycin A inhibited the proton release, and myxothiazol almost completely abolished it. PMID- 3032926 TI - Electron spin resonance spectra of free radicals formed in the reaction of metmyoglobins with ethylhydroperoxide. AB - Free radicals of myoglobins were measured at room temperature with an ESR spectrometer equipped with a flow apparatus. When horse heart MetMb was mixed with an equimolar amount of ethyl hydroperoxide (EtOOH), a well resolved ESR spectrum with 6 lines and a shoulder was observed. It reached a maximum in a few seconds and decayed with a half-life of about 10 s when the final concentrations of MetMb and EtOOH were 200 microM. This decay rate was the same at a MetMb concentration of 50 microM. The maximum molar radical concentration amounted to about half of the total myoglobin. In the case of sperm whale myoglobin, a similar 6-line spectrum reached a maximum in 1 s and decayed with a half-life of a few seconds. In this case, however, a small and poorly resolved doublet spectrum remained, the half-life of which was about 8 min. An effect of O2 on the signal decay was evident for horse heart myoglobin, but not for sperm whale myoglobin. PMID- 3032927 TI - Cyclic GMP regulation of the plasma membrane (Ca2+-Mg2+)ATPase in vascular smooth muscle. AB - Plasma membrane (Ca2+-Mg2+)ATPase purified from bovine aortic microsomes by calmodulin affinity chromatography was incorporated into soybean phospholipid liposomes. In the reconstituted proteoliposomes, a protein corresponding to the ATPase was phosphorylated by [gamma-32P]ATP in the presence of cGMP and cGMP dependent protein kinase. Both the affinity for Ca2+ and the maximum Ca2+ uptake activity by the proteoliposomes were increased by the cGMP-dependent phosphorylation, and there was good parallelism between the Ca2+-uptake rate and the extent of phosphorylation. These results strongly suggest that the Ca2+ transport ATPase of the vascular smooth muscle plasma membrane is regulated through its cGMP-dependent phosphorylation. PMID- 3032928 TI - The plasma membrane ATPase of Neurospora: a proton-pumping electroenzyme. AB - Probably the best marker enzyme for plasma membranes of eukaryotic cells is a magnesium-dependent, vanadate-inhibited ATPase whose primary function is the transmembrane transport of cations. In animal cells, different species of the enzyme transport different cations: sodium ions released in unequal exchange for potassium ions, calcium ions extruded alone (perhaps), or protons secreted in equal exchange for potassium ions. But in plants and fungi only proton secretion has been clearly demonstrated. A useful model cell for studying the proton secreting ATPase has been the ascomycete fungus Neurospora, in which the enzyme drives an outward current of protons that can exceed 50 microA/cm2 and can support membrane potentials greater than 300 mV. Both thermodynamic and kinetic studies have shown that the proton-pumping ATPase of Neurospora normally transports only a single proton for each ATP molecule split; and kinetic modelling studies have suggested (contrary to conventional assumptions) that the fast steps in the overall reaction are transmembrane transit of the proton and its dissociation following transport, while the slow steps are the binding of protons and/or ATP. The primary structure of the Neurospora enzyme, recently deduced by gene sequencing, is very close to that of the yeast (Saccharomyces) enzyme, and the hydropathic patterns for both closely resemble those for the animal-cell plasma-membrane ATPases. All of these enzymes appear to have 6-10 membrane-spanning alpha-helices, plus a large cytoplasmic headgroup which bears the catalytic nucleotide-binding site. Structural data, taken together with the electrical-kinetic behavior, suggest that the catalytic headgroup functions as an energized gate for protons. From a geometric point of view, action of such a gate would transfer the membrane field across the "transported" ion, rather than vice versa. PMID- 3032929 TI - Effect of succinate on mitochondrial lipid peroxidation. 2. The protective effect of succinate against functional and structural changes induced by lipid peroxidation. AB - The damaging effects of ADP/Fe/NADPH-induced lipid peroxidation were studied on the enzymes and membranes of rat liver mitochondria. Succinate, an inhibitor of mitochondrial lipid peroxidation, prevented or delayed most of the damage caused by the peroxidation on different mitochondrial structures and functions. There were marked abnormalities on the electrophoretic pattern of mitochondrial proteins during the course of lipid peroxidation. The disappearance of particular polypeptide bands and the accumulation of high-molecular-weight aggregates could be observed. Succinate was found to delay these effects. As a consequence of lipid peroxidation the succinate oxidase activity of mitochondria was decreased. The succinate dehydrogenase enzyme and the component(s) of the respiratory chain were inactivated. Succinate prevented the inactivation of succinate dehydrogenase but did not protect the other components of terminal oxidation chain. From the matrix enzymes the glutamate dehydrogenase retained its full activity but the NADP-linked isocitrate dehydrogenase was inactivated. The mitochondrial membranes became permeable to large protein molecules. Succinate prevented the inactivation of isocitrate dehydrogenase and delayed the release of protein molecules from mitochondria. PMID- 3032931 TI - 5-Fluorouracil augmentation of dihydrofolate reductase RNA containing contiguous exon and intron sequences in KB7B cells. AB - Quantitative S1 nuclease mapping studies were performed with uniformly labeled RNA probes, containing contiguous dihydrofolate reductase exon and intron sequences, and total RNA isolated from KB7B cells exposed to 5-fluorouracil for 5 days. Dihydrofolate reductase RNA containing both exon 1 and intron I, or exon 5 and a portion of intron V, increased up to 5-fold in cells grown in the presence of 2.0 to 3.0 microM 5-fluorouracil. Dihydrofolate reductase RNA containing exon 1 or exon 5, but lacking intron I or intron V, respectively, increased 2-fold in cells grown in the presence of 0.65 to 3.0 microM 5-fluorouracil. Primer extension analysis and S1 mapping studies revealed two major transcriptional start sites at positions -72 and -69 and minor start sites upstream from position -183, for dihydrofolate reductase RNA isolated from methotrexate-resistant KB7B cells. The results of these studies demonstrate that 5-fluorouracil alters the metabolism of dihydrofolate reductase precursor mRNA and/or processing intermediates. PMID- 3032930 TI - Interaction of photosystem I-derived protons with the water-splitting enzyme complex. Evidence for localized domains. AB - The induction of millisecond delayed fluorescence mediated by PS I-dependent proton pumping has been used as an indicator of the time course with which those protons equilibrate with sites on the oxygen-evolving enzyme complex (Bowes, J. M., and Crofts, A. R. (1978). Z. Naturforsch. 33C, 271-275). We found that the induction curves were retarded by a reversible exposure of non-energized thylakoids to low concentrations of the uncoupler, desaspidin, at alkaline, but not at neutral, pH. The induction curves were not retarded by increasing the buffering capacity of the thylakoid lumen with Tricine, and were inhibited by the energy transfer inhibitors, dicyclohexylcarbodiimide (DCCD) and triphenyltin chloride (TPT). These data suggest that the catalytic site of the water-splitting complex is located in proton-sequestering membrane domains, rather than at the lumen-exposed inner membrane surface, protons released during PS I-mediated electron transport might equilibrate with protonatable sites on the oxygen evolving complex without passing through the lumen, and those protons may travel over specific conducting pathways which can be blocked by DCCD and TPT. PMID- 3032932 TI - Subunit VII, the ubiquinone-binding protein, of the cytochrome b-c1 complex of yeast mitochondria is involved in electron transport at center o and faces the matrix side of the membrane. AB - The functional role and topographical orientation in the inner membrane of subunit VII, the ubiquinone-binding protein, of the cytochrome b-c1 complex of yeast mitochondria has been investigated. The apparent molecular weight of this subunit on sodium dodecyl sulfate-urea gels was calculated to be 15,500, while its amino acid composition was similar to that of the Q-binding proteins present in the cytochrome b-c1 complexes isolated from both beef heart and yeast mitochondria. The specific antibody obtained against subunit VII inhibited 30-47% of the ubiquinol-cytochrome c reductase activity in the isolated cytochrome b-c1 complex and in submitochondrial particles but had no effect on cytochrome c reductase activity in mitoplasts, mitochondria from which the outer membrane has been removed. Furthermore, the antibody against subunit VII strongly inhibited (74%) the reduction of cytochrome b by succinate in the presence of antimycin, an inhibitor of center i, but had no effect on cytochrome b reduction in the presence of myxothiazol, an inhibitor of center o. These results suggest that subunit VII, the Q-binding protein, is involved in electron transport at center o of the cytochrome b-c1 complex of the respiratory chain and that subunit VII is localized facing the matrix side of the inner mitochondrial membrane. PMID- 3032934 TI - Purification of atrial natriuretic peptide receptor from bovine lung. Evidence for a disulfide-linked subunit structure. AB - The receptor for atrial natriuretic peptide (ANP) was purified to apparent homogeneity from bovine lung by a combination of detergent extraction, ammonium sulfate precipitation, and affinity chromatography on ANP-Affi-Gel 10. The Mr of the purified receptor is about 140,000 as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis. After reduction, the protein migrated as a single band with an Mr near 70,000. NH2-terminal sequence analysis of the purified material revealed only one sequence, indicating that the ANP receptor is composed of two probably identical subunits held together by disulfide bond(s), although it remains possible that one of the subunits is blocked at the NH2 terminus. Antibody was produced to the nonreduced Mr = 140,000 species and shown to interact with detergent-solubilized forms of the lung and kidney ANP receptor. PMID- 3032933 TI - Thrombin- and nucleotide-activated phosphatidylinositol 4,5-bisphosphate phospholipase C in human platelet membranes. AB - Thrombin, nucleotides, and chelators elicited a phosphatidylinositol 4,5 bisphosphate (PtdIns-P2) phospholipase C activity that was associated with human platelet membranes. Both alpha- and gamma-thrombin enhanced phospholipase C activity, whereas active site-inhibited alpha-thrombin did not stimulate PtdIns P2 hydrolysis. PtdIns-P2 phospholipase C was also activated by nucleoside triphosphates, citrate, EDTA, and NaF. Magnesium was an inhibitor of PtdIns-P2 hydrolysis stimulated by nucleotides and chelators. Only PtdIns-P2 was degraded by the phospholipase C activated by alpha-thrombin, nucleotides, and chelators. The soluble fraction phospholipase C activity was also stimulated at low protein concentrations by nucleotides; however, soluble fraction phospholipase C activity cleaved both PtdIns-P2 and phosphatidylinositol 4-phosphate and was inhibited by chelators, suggesting the presence of a different enzyme in this compartment. The pH optimum for the membrane-associated phospholipase C in the presence of alpha thrombin or nucleotides was 6.0, and the PtdIns-P2 phospholipase C was inhibited by neomycin and high detergent concentrations. Guanine nucleotides did not synergistically activate phospholipase C in the presence of alpha-thrombin. The characteristics of the membrane-associated PtdIns-P2 phospholipase C suggest that this enzyme is involved in platelet activation by the low-affinity alpha- or gamma-thrombin-dependent pathway. PMID- 3032935 TI - Isolation and characterization of adenosine kinase from Leishmania donovani. AB - Adenosine kinase (ATP:adenosine 5'-phosphotransferase, EC 2.7.1.20) has been purified 3250-fold from Leishmania donovani promastigotes using ion-exchange, gel filtration, and affinity chromatography techniques. Both native and sodium dodecyl sulfate-gel electrophoresis of the enzyme revealed a single polypeptide of around 38,000 molecular weight. Biophysical and biochemical analyses of the enzyme reveal unique characteristics different from those of adenosine kinases from other eukaryotic sources. The isoelectric pH of the enzyme is 8.8. In native acrylamide gels the enzyme moves with an RF of about 0.62. The enzyme displays a maximum activity at pH between 7.5 and 8.5 and is dependent upon an optimum ATP/Mg2+ ratio. ATP at high concentration inhibits the reaction. Adenosine and Mg2+ are not inhibitory. EDTA completely knocks off the activity. Enzyme activity is dependent upon the presence of active thiol group(s) at or near the active center. Under a defined set of conditions the enzyme exhibited an apparent Km for adenosine and ATP of 33 and 50 microM, respectively. Of the nucleoside triphosphates tested ATP and GTP were the most effective phosphate donors. Marginal inhibition of activity was detected with other nucleosides as competitors. However, adenosine analogs, such as 7-deaza-adenosine (tubercidin) and 6-methylmercaptopurine riboside at very low concentrations, were found to be excellent inhibitors and substrates as well. S-Adenosylhomocysteine does not inhibit the reaction even at very high concentration. PMID- 3032936 TI - Phosphatidic acid may stimulate membrane receptors mediating adenylate cyclase inhibition and phospholipid breakdown in 3T3 fibroblasts. AB - Incubation of 3T3 fibroblasts with phosphatidic acid (PA) from egg lecithin or with thrombin resulted in decreases in cellular cAMP due to inhibition of adenylate cyclase, in rapid increases in inositol 1,4,5-tris-,1,4-bis-, and 1 monophosphates probably due to activation of phospholipase C, and in arachidonic acid release. Synthetic PAs consisting of unsaturated fatty acid diesters were as effective as PA from egg lecithin, whereas PAs with saturated fatty acids were only slightly effective and antagonized the effect of active PAs selectively, despite the fact that both types of PA analogues (sodium salts) were apparently dissolved in the incubation medium. PA-induced decreases in cAMP were not affected by omission of Ca2+ from incubation medium but were abolished by prior exposure of cells to islet-activating protein (pertussis toxin). This islet activating protein treatment of cells was without effect on PA- or thrombin induced generation of inositol phosphates. Thus, PA-induced inhibition of adenylate cyclase was (but activation of phospholipase C was not) mediated by an islet-activating protein substrate GTP-binding protein. Homologous desensitization was observed with thrombin-, bradykinin-, and PA-induced decreases in cAMP in 3T3 cells; prior exposure of the cells to any one of these agents abolished or greatly diminished the subsequent response to the same agent but did not affect the responses to others. The effects of PA were cell-specific; it failed to decrease cAMP in rabbit platelets in which labeled PA rapidly increasing in response to thrombin or A23187 was mostly outside the cells. Based on these results, it is proposed that PA interacts with its own specific membrane receptors, thereby triggering multiple effector systems in 3T3 cells. PMID- 3032937 TI - Binding and receptor-mediated endocytosis of erythropoietin in Friend virus infected erythroid cells. AB - The binding of labeled erythropoietin (EP) to cell surface receptors and subsequent processing of the hormone within the cell was studied in erythroid cells procured from the spleens of mice infected with the anemia strain of Friend virus. These immature erythroid cells respond to EP in culture to differentiate into reticulocytes and erythrocytes. Radiolabeled EP (both iodinated and tritiated) binds to 800-1000 cell surface receptors on these cells at 4 degrees C. Using 125I-EP, we found that 300 of these cell surface receptors have a higher affinity for EP (Kd = 0.09 nM) than the remaining receptors (Kd = 0.57 nM). The number of molecules of EP bound per cell increased about 2-fold when binding was carried out at 37 degrees C. Treatment of the cell surface with pronase or removal of surface-bound EP with a low pH wash revealed that radiolabeled EP is internalized by the cells at 37 degrees C. Pulse chase experiments showed that degradation products of radiolabeled EP are released into the medium with a corresponding loss of label from the interior of the cell. Inhibitors of lysosomal function greatly reduced this degradation of 125I-EP. Since 180 of the 300 high affinity receptors and very few of the low affinity receptors are occupied at the concentration of EP which elicits the maximum biological response in these cells, we suggest that interaction of EP with the high affinity receptors are necessary for the full biological effect of the hormone. A different murine erythroleukemia cell line which does not differentiate in response to EP was found to have only the lower affinity binding sites for the hormone. PMID- 3032938 TI - Mechanisms of accelerated purine nucleotide synthesis in human fibroblasts with superactive phosphoribosylpyrophosphate synthetases. AB - Concentrations and rates of synthesis of phosphoribosylpyrophosphate (PP-Rib-P) and purine nucleotides were compared in fibroblasts cultured from 5 males with PP Rib-P synthetase superactivity, 3 normal individuals, and 2 children with severe hypoxanthine-guanine phosphoribosyltransferase deficiency. Although all cell strains with PP-Rib-P synthetase superactivity showed increased PP-Rib-P concentration and generation, increased rates of PP-Rib-P-dependent purine synthetic pathways, and increased purine and pyrimidine nucleoside triphosphate concentrations, two subgroups were discernible. Three fibroblast strains with isolated catalytic defects in PP-Rib-P synthetase showed milder increases in PP Rib-P concentration (2.5-fold normal) and generation (1.6- to 2.1-fold) and in rates of purine synthesis de novo (1.6- to 2.2-fold) and purine nucleoside triphosphate pools (1.5-fold) than did cells from 2 individuals with combined kinetic defects in PP-Rib-P synthetase, both with purine nucleotide inhibitor resistance. Values for these processes in the latter two strains were, respectively, 5- to 6-fold, 2.6- to 3.2-fold, 4- to 7-fold, and 1.7- to 2.2-fold those of normal cells. In contrast to cells with catalytic defects, these cells also excreted an abnormally high proportion of labeled purines and resisted purine base-mediated inhibition of PP-Rib-P and purine nucleotide synthesis. Hypoxanthine-guanine phosphoribosyltransferase-deficient cells showed normal regulation of PP-Rib-P synthesis and normal nucleoside triphosphate pools despite increased rates of purine synthesis de novo and of purine excretion. Cells with PP-Rib-P synthetase superactivity thus synthesize purine nucleotides at increased rates as a consequence of increased PP-Rib-P production, despite increased purine nucleotide concentrations. These and additional findings provide evidence that regulation of purine synthesis de novo is effected at both the PP-Rib-P synthetase and amidophosphoribosyltransferase reactions. PMID- 3032939 TI - Diacylglycerol accumulation and superoxide anion production in stimulated human neutrophils. AB - Exogenous diacylglycerols stimulate neutrophil superoxide anion production, suggesting that endogenous diacylglycerols may function as second messengers for this biological response. We have measured the diacylglycerol mass in human neutrophils stimulated by fMet-Leu-Phe, ionomycin, and concanavalin A and have correlated the kinetics and magnitude of the diacylglycerol response with those for superoxide anion production. For each stimulus, no increase in diacylglycerol mass was detected prior to the onset of superoxide anion generation. However, large sustained increases in diacylglycerol concentration (260-2000% of basal levels) occurred in parallel with the rise in superoxide anion. The cessation or continuation of diacylglycerol accumulation and superoxide anion production also correlated. The diacylglycerol response was proportional to the stimulus concentration and correlated with the concentration dependence for superoxide anion. Pretreatment of neutrophils with cytochalasin B enhanced both superoxide anion and diacylglycerol responses with all three stimuli. These data support the hypothesis that diacylglycerol functions as a modulator of superoxide anion generation causing a sustained or augmented respiratory burst. PMID- 3032940 TI - Crystal structure of a cyclic AMP-independent mutant of catabolite gene activator protein. AB - Escherichia coli NCR91 synthesizes a mutant form of catabolite gene activator protein (CAP) in which alanine 144 is replaced by threonine. This mutant, which also lacks adenylate cyclase activity, has a CAP phenotype; in the absence of cAMP it is able to express genes that normally require cAMP. CAP91 has been purified and crystallized with cAMP under the same conditions as used to crystallize the wild type CAP X cAMP complex. X-ray diffraction data were measured to 2.4-A resolution and the CAP91 structure was determined using initial model phases from the wild type structure. A difference Fourier map calculated between CAP91 and wild type showed the 2 alanine to threonine sequence changes in the dimer and also a change in orientation of cysteine 178 in one of the subunits. The CAP91 coordinates were refined by restrained least squares to an R factor of 0.186. Differences in the atomic positions of the wild type and mutant protein structures were analyzed by a vector averaging technique. There were small changes that included concerted motions in the small domains, in the hinge between the two domains and in an adjacent loop between beta-strands 4 and 5. The mutation at residue 144 apparently causes changes in the position of some protein atoms that are distal to the mutation site. PMID- 3032941 TI - Characteristics of a pure endogenous activator of the gastric H+,K+-ATPase system. Evaluation of the role as a possible intracellular regulator. AB - An endogenous activator capable of stimulating the gastric H+,K+-ATPase activity has been purified to homogeneity from dog and pig gastric cells and found to be a dimer of two identical 40-kDa subunits in the active state. Identical nature of the activator monomers was revealed by the detection of lysine as the sole N terminal amino acid. The activator from one species can stimulate the H+,K+ ATPase from another species and vice versa. Such cross-activation is consistent with the striking similarities in the amino acid composition between the two species, suggesting considerable homology in the activator molecules from different species. The activator exhibited several unique features during modulation of the H+,K+-ATPase reaction. It appreciably enhances affinity of the H+,K+-ATPase for K+, known to increase turnover of the enzyme. To complement this K+ affinity, the activator also enhances ability of the H+,K+-ATPase to generate more transition state (E*.ATP) complex by increasing the entropy of activation (delta S++) of the system as revealed from an Arrhenius plot of the data on temperature activation. In addition, the activator shows both positive cooperativity and strong inhibition, depending on its concentration. Thus, up to the ratio of the H+,K+-ATPase and activator of about 1:2 (on the protein basis), the activator shows sigmoidal activation (Hill coefficient = 4.5), but beyond such concentration a strong inhibition was observed. Finally, Ca2+ at low (2-4 microM) concentration strongly inhibits the activator-stimulated H+,K+-ATPase. It is proposed that the activator may be acting as a link in the signal transducing cascade system between the intracellular second messenger (Ca2+) and the physiological response (gastric H+ transport). PMID- 3032942 TI - Altered signal transduction in erbB-transformed cells. Implication of enhanced inositol phospholipid metabolism in erbB-induced transformation. AB - To clarify the signal transduction mechanism of the erbB gene (virus oncogene) products leading to cell growth and transformation, the alteration of signal transduction induced by enhanced inositol phospholipid metabolism was studied in chick embryo fibroblast cells (CEF cells) transformed by gag-fused erbB gene carrying virus (GEV cells). The incorporations of 32P into phosphatidylinositol 4 phosphate (PIP) and phosphatidylinositol 4,5-bisphosphate were markedly increased in GEV cells. In GEV cells, the activities of lipid kinases such as phosphatidylinositol (PI), PIP, and diacylglycerol (DG) kinases were also increased. The activities of other important enzymes involved in inositol phospholipid metabolism, such as CDP-DG:myo-inositol transferase and phospholipase C, were not changed in GEV cells. Increased inositol phospholipid metabolism might lead to the production of second messengers, such as 1,2-DG and inositol 1,4,5-trisphosphate. Indeed, the 1,2-DG content was also increased in GEV cells. Moreover, the activity of protein kinase C (the Ca2+/phospholipid dependent enzyme), which should be stimulated by 1,2-DG, was elevated in GEV cells; the protein kinase C activity in the membrane fraction of GEV cells was especially high. When CEF cells were treated with tetradecanoylphorbol acetate, protein kinase C activator, plus Ca2+ ionophore, [3H]thymidine incorporation was markedly stimulated, and maximal stimulation was observed with 1 nM Ca2+ ionophore A23187 plus 100 nM TPA. On the other hand, when GEV cells were treated with TPA plus Ca2+ ionophore A23187, [3H]thymidine incorporation was consistently inhibited. Next, studies were made to determine whether the erbB gene product itself had kinase activity on PI, PIP, and DG after membranes were mildly solubilized with Triton X-100 to prevent inactivation of these kinases. Immunoprecipitates of a GEV cell lysate with antisera that reacted with the erbB gene product had PI kinase activity, whereas no activity was detected in those of lysates of uninfected CEF cells. However, the activity was very weak compared with the total cellular activity. No difference in the PIP and DG kinase activities of immunoprecipitates of cell lysates of uninfected CEF cells and GEV cells was observed. These results suggest that the erbB gene product enhances inositol phospholipid metabolism and subsequent signal transduction, but that the erbB gene product is not involved directly in lipid kinases, although it is closely associated with lipid kinase. PMID- 3032943 TI - Primary structures of bovine elastin a, b, and c deduced from the sequences of cDNA clones. AB - Nucleotide sequence analysis of cDNA clones for bovine elastin revealed the occurrence of three mRNAs for elastin in fetal calf nuchal ligament, encoding three forms of elastins (a, b, and c, of 747, 733, and 713 amino acid residues, respectively). These forms arise as the result of the presence, at a single position, of 102 additional nucleotides in the mRNA for elastin a and of 60 of these nucleotides in the mRNA for elastin b as compared to the mRNA for elastin c. As expected, most lysines occur in pairs, separated by two or three small amino acid residues. However, at two places, lysines occur in groups of three. The occurrence of a group of three lysines followed by a hydrophobic residue (lysine 400, 404, and 407) offers an explanation for the formation of lysinonorleucine. The alignment of amino acid sequences of porcine tropoelastin tryptic peptides with the sequence for bovine elastin a results in the ordering of these tryptic peptides. The analysis of the complete primary structures of elastin a, b, and c provides further insight into the structure-function relations of elastin. PMID- 3032944 TI - Purification of the putative hormone-sensitive cyclic AMP phosphodiesterase from rat adipose tissue using a derivative of cilostamide as a novel affinity ligand. AB - A "low Km" cAMP phosphodiesterase with properties of a peripheral membrane protein accounts for approximately 90% of total cAMP phosphodiesterase activity in particulate (100,000 X g) fractions from rat fat cells. Incubation of fat cells with insulin for 10 min increased particulate (but not soluble) cAMP phosphodiesterase activity, with a maximum increase (approximately 100%) at 1 nM insulin. Most of the increase in activity was retained after solubilization (with non-ionic detergent and NaBr) and partial purification (approximately 20-fold) on DEAE-Sephacel. The solubilized enzyme from adipose tissue was purified approximately 65,000-fold to apparent homogeneity (yield approximately 20%) by chromatography on DEAE-Sephacel and Sephadex G-200 and affinity chromatography on aminoethyl agarose conjugated with the N-(2-isothiocyanato)ethyl derivative of the phosphodiesterase inhibitor cilostamide (OPC 3689). A 63,800 +/- 200-Da polypeptide (accounting for greater than 90% of the protein eluted from the affinity column) was identified by polyacrylamide gel electrophoresis in sodium dodecyl sulfate (with or without reduction). Enzyme activity was associated with the single protein band after electrophoresis under nondenaturing conditions. On gel permeation, Mr(app) was 100,000-110,000, suggesting that the holoenzyme is a dimer. A pI of 4.9-5.0 was estimated by isoelectric focusing. At 30 degrees C, the purified enzyme hydrolyzed both cAMP and cGMP with normal Michaelis-Menten kinetics; the pH optimum was 7.5. The Km(app) for cAMP was 0.38 microM and Vmax, 8.5 mumol/min/mg; for cGMP, Km(app) was 0.28 microM and Vmax, 2.0 mumol/min/mg. cGMP competitively inhibited cAMP hydrolysis with a Ki of approximately 0.15 microM. The enzyme was also inhibited by several OPC derivatives and "cardiotonic" drugs, but not by RO 20-1724. It was very sensitive to inhibition by agents which covalently modify protein sulfhydryls, but not by diisopropyl fluorophosphate. The activation by insulin and other findings indicate that the purified enzyme, which seems to belong to a subtype of low Km cAMP phosphodiesterases that is specifically and potently inhibited by cGMP, cilostamide, other OPC derivatives, and certain cardiotonic drugs, is likely to account for the hormone-sensitive particulate low Km cAMP phosphodiesterase activity of rat adipocytes. PMID- 3032946 TI - Purification of the Ca2+-dependent proteinase inhibitor from bovine cardiac muscle and its interaction with the millimolar Ca2+-dependent proteinase. AB - A protein inhibitor of the Ca2+-dependent proteinase has been purified from bovine cardiac muscle by using the following steps in succession: salting out 17,600 X gmax supernatants from muscle homogenates in 50 mM Tris acetate, pH 7.5, 4 mM EDTA between 25 and 65% ammonium sulfate saturation; eluting between 25 and 120 mM KCl from a DEAE-cellulose column at pH 7.5; salting out between 30 and 60% ammonium sulfate saturation; Ultrogel-22 gel permeation chromatography at pH 7.5; heating to 80 degrees C followed by immediate cooling to 0 degree C; 6% agarose gel permeation chromatography in 4 M urea, pH 7.5; and elution from a phenyl Sepharose hydrophobic column between 0.7 and 0.5 M ammonium sulfate. Approximately 1.16-1.69 mg of purified Ca2+-dependent proteinase inhibitor are obtained from 1 kg of bovine cardiac muscle, fresh weight. Bovine cardiac Ca2+ dependent proteinase inhibitor has an Mr of 115,000 as measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, a pI of 4.85-4.95, very little alpha helical structure, a very low specific absorbance of 1.647 (A1% 280), and very low contents of histidine, tryptophan, phenylalanine, and tyrosine. Bovine cardiac Ca2+-dependent proteinase inhibitor probably contains a single polypeptide chain in nondenaturing solvents. One 115-kDa inhibitor polypeptide inactivates 10 110-kDa millimolar Ca2+-requiring proteinase (millimolar Ca2+ dependent proteinase) molecules in assays of purified proteins. Inhibition of millimolar proteinase by the proteinase inhibitor did not change in the pH range 6.2-8.6. The inhibitor requires Ca2+ to bind to millimolar Ca2+-dependent proteinase. The Ca2+ concentration required for one-half-maximum binding of millimolar Ca2+-dependent proteinase to the inhibitor was 0.53 mM, compared with a Ca2+ concentration of 0.92 mM required for one-half maximum activity of millimolar Ca2+-dependent proteinase in the absence of the proteinase inhibitor. Unless millimolar Ca2+-dependent proteinase is located subcellularly in a different place than the proteinase inhibitor or unless the proteinase/inhibitor interaction is regulated, millimolar proteinase could never be active in situ. PMID- 3032945 TI - A single precursor protein for two separable mitochondrial enzymes in Neurospora crassa. AB - The arg-6 locus of Neurospora crassa encodes two early enzymes of the arginine biosynthetic pathway, acetylglutamate kinase and acetylglutamyl-phosphate reductase. Previous genetic and biochemical analyses of this locus and its products showed that: 1) strains carrying polar nonsense mutations in the acetylglutamate kinase gene lacked both enzyme activities (Davis, R.H., and Weiss, R.L. (1983) Mol. Gen. Genet. 192, 46-50), and 2) the proteins isolated from mitochondria were completely separable (Wandinger-Ness, A., Wolf, E.C., Weiss, R.L., and Davis, R.H. (1985) J. Biol. Chem. 260,5974-5978). These data suggested that the two enzymes were initially synthesized as a single precursor which was subsequently cleaved into two distinct polypeptides. We report here the identification of a high molecular weight protein, synthesized in vitro from isolated N. crassa RNA, that contains sequences corresponding to acetylglutamate kinase as well as acetylglutamyl-phosphate reductase. An analogous precursor was identified in vivo by pulse-labeling experiments. The precursor was similar to other mitochondrial precursors in that its uptake and processing in vivo was rapid and required an intact mitochondrial electrochemical gradient. This represents the first report of a bifunctional protein precursor which gives rise to two mitochondrial enzymes. PMID- 3032947 TI - The activation of human skin fibroblast procollagenase. Sequence identification of the major conversion products. AB - Human skin collagenase is secreted by cultured fibroblasts in a proenzyme form and can be activated to a catalytically competent enzyme by a number of processes. All modes of activation studied lead to conversion of the proenzyme to a stable 42-kDa active enzyme, concomitant with removal of an 81-amino acid peptide from the amino-terminal end of the molecule. The sequence of events leading to the formation of this enzyme form has been determined by analyzing the primary structure of the conversion intermediates. Trypsin-induced activation of procollagenase occurs as a result of the initial cleavage of the peptide bond between Arg-55 and Asn-56, generating a major intermediate of 46 kDa. Treatment of the proenzyme with organomercurials, which have no intrinsic ability to cleave peptide bonds, initially results in activation of the enzyme without loss of molecular weight. This is followed by conversion to two lower molecular weight species of 44 and 42 kDa, the latter corresponding to the stable active enzyme form. The final cleavage producing this form of collagenase is not restricted to a single polypeptide bond but can occur on the amino-terminal side of any one of three contiguous hydrophobic residues, Phe-100, Val-101, Leu-102. The data suggest that both trypsin and organomercurials activate procollagenase by initiating an intramolecular autoproteolytic reaction resulting in the formation of a stable 42-kDa active enzyme species. PMID- 3032948 TI - Characterization of a second gene product related to rabbit cytochrome P-450 1. AB - A cDNA, p1-88, was cloned from a library constructed using rabbit liver mRNA. Sequence analysis indicates that p1-88 is highly similar (congruent to 95%) to the cDNA, p1-8, that encodes rabbit liver cytochrome P-450 1 and that had been isolated from the same library. The predicted amino acid sequence of the protein encoded by p1-88, P-450 IIC4, differs at 25 of 487 amino acids from that encoded by p1-8. P-450 IIC4 was synthesized in vitro using rabbit reticulocyte lysate primed with RNA transcribed from the coding sequence of p1-88 using a bacteriophage T7 RNA polymerase/promoter system. P-450 IIC4 reacts with two monoclonal antibodies that recognize P-450 1 and exhibits the same relative electrophoretic mobility as P-450 1. In contrast, the reactivity of a third monoclonal antibody recognizing P-450 1, 1F11, toward P-450 IIC4 synthesized in vitro is greatly diminished. The latter antibody extensively inhibits hepatic progesterone 21-hydroxylase activity and recognizes phenotypic differences among rabbits in the microsomal concentration of P-450 1. This difference in the immunoreactivity of P-450 IIC4 and P-450 1 with the 1F11 antibody suggests that P 450 IIC4 does not contribute significantly to hepatic progesterone 21-hydroxylase activity. S1 nuclease mapping demonstrates that the expression of mRNAs corresponding to p1-88 are expressed to equivalent extents in rabbits exhibiting high and low expression of mRNAs corresponding to p1-8. Thus, P-450 1 differs from the protein encoded by p1-88, in its regulation, immunoreactivity, and by inference its catalytic properties although the amino acid sequences of P-450 1 and P-450 IIC4 are highly similar (congruent to 95%). PMID- 3032949 TI - Structure and organization of the microsomal xenobiotic epoxide hydrolase gene. AB - The gene for the microsomal xenobiotic rat liver epoxide hydrolase has been isolated and characterized. Clones were obtained from a Wistar Furth Charon 35 genomic library by hybridization with a full-length epoxide hydrolase cDNA. The gene for the xenobiotic epoxide hydrolase is approximately 16 kilobases in length and consists of 9 exons ranging in size from 109 to 420 base pairs and 8 intervening sequences, the largest of which is 3.2 kilobases. S1-nuclease mapping, primer extension studies, and sequence analysis were used to determine the 5' cap site and the size of the first exon (170 base pairs). Regulatory sequences analogous to TATA, CCAAT, and core enhancer sequences were noted in the 5'-flanking region of the gene. The cDNA and gene for epoxide hydrolase displayed nucleotide sequence identity although they were isolated from different rat strains. Also, Southern blot analysis of restricted liver DNA from inbred Fischer 344 and Wistar Furth rat strains, and outbred Sprague-Dawley rats indicated a high degree of structural similarity for the epoxide hydrolase gene within these three strains. Only a single functional epoxide hydrolase gene was identified and no evidence of hybridization to the genes for the microsomal cholesterol epoxide hydrolase or the cytosolic epoxide hydrolase was observed. However, a pseudogene for the microsomal xenobiotic epoxide hydrolase was isolated and characterized from the genomic library. PMID- 3032950 TI - Human fibroblast collagenase contains an amino acid sequence homologous to the zinc-binding site of Serratia protease. AB - Analysis of sequence alignments between the recently published sequence of human fibroblast collagenase and a computer file of published protease sequences has revealed a previously unrecognized homology to the 11 amino acids flanking the zinc-binding site of Serratia protease, a bacterial metalloprotease. There is also strong homology among several bacterial metalloproteases at this site. This finding implies that zinc binding by many proteins may have structural requirements which are apparent even in primary structure and which have been evolutionarily conserved or convergently evolved. This consensus sequence could be used as a marker for recognizing other members of the metalloprotease family. PMID- 3032951 TI - ATP induces conformational changes in mitochondrial cytochrome c oxidase. Effect on the cytochrome c binding site. AB - ATP influences the kinetics of electron transfer from cytochrome c to mitochondrial oxidase both in the membrane-embedded and detergent-solubilized forms of the enzyme. The most relevant effect is on the so-called "high affinity" binding site for cytochrome c which can be converted to "low affinity" by millimolar concentrations of ATP (Ferguson-Miller, S., Brautigan, D. L., and Margoliash, E. (1976) J. Biol. Chem. 251, 1104-1115). This phenomenon is characterized at the molecular level by the following features. ATP triggers a conformational change on the water-exposed surface of cytochrome c oxidase; in this process, carboxyl groups forming the cluster of negative charges responsible for binding cytochrome c change their accessibility to water-soluble protein modifier reagents; as a consequence the electrostatic field that controls the enzyme-substrate interaction is altered and cytochrome c appears to bind differently to oxidase; photolabeling experiments with the enzyme from bovine heart and other eukaryotic sources show that ATP cross-links specifically to the cytoplasmic subunits IV and VIII. Taken together, these data indicate that ATP can, at physiological concentration, bind to cytochrome c oxidase and induce an allosteric conformational change, thus affecting the interaction of the enzyme with cytochrome c. These findings raise the possibility that the oxidase activity may be influenced by the cell environment via cytoplasmic subunit-mediated interactions. PMID- 3032952 TI - DNA sequences of the cysB regions of Salmonella typhimurium and Escherichia coli. AB - Nucleotide sequences of the cysB region of Salmonella typhimurium and Escherichia coli have been determined and compared. A total of 1759 nucleotides were sequenced in S. typhimurium and 1840 in E. coli. Both contain a 972-nucleotide open reading frame identified as the coding region for the cysB regulatory protein on the basis of sequence homology and by comparison of the deduced amino acid sequences with known physicochemical properties of this protein. The DNA sequence identity for the cysB coding region in the two species is 80.5%. The deduced amino acid sequences are 95% identical. The predicted cysB polypeptide molecular weights are 36,013 for S. typhimurium and 36,150 for E. coli. For both proteins a helix-turn-helix region similar to that found in other DNA-binding proteins is predicted from the deduced amino acid sequence. Sequences upstream to cysB contain open reading frames which represent the carboxyl-terminal end of the topA gene product, DNA topoisomerase I. A pattern of highly conserved nucleotide sequences in the 151 nucleotides immediately preceding the cysB initiator codon in both species suggests that this region may contain multiple signals for the regulation of cysB expression. PMID- 3032953 TI - Purification of the cysB protein from Salmonella typhimurium. AB - Using an assay based on polypeptide mobility in one- and two-dimensional polyacrylamide electropherograms, we purified the Salmonella typhimurium cysB protein from an Escherichia coli strain carrying plasmid pGBK14, in which the S. typhimurium cysB gene is under transcriptional control of the strong promoter PL from phage lambda. cysB protein constitutes approximately 4% of total soluble protein in such cells and was obtained with good yield at a purity of 85% after precipitation of nucleic acids with streptomycin sulfate, ammonium sulfate fractionation, and hydrophobic chromatography on methyl agarose. Material with 95% purity was obtained with poor yield by ion-exchange chromatography of native protein and with good yield by size exclusion chromatography of denatured protein in sodium dodecyl sulfate. Physicochemical studies of the purified cysB protein show that it is a tetramer of identical 36-kDa subunits with a pI, amino acid composition, amino-terminal sequence, and carboxyl-terminal amino acid content identical to those known from previous studies or deduced from the cysB DNA sequence. Although O-acetyl-L-serine is believed to bind to cysB protein to form a complex that stimulates transcription of genes of the cysteine regulon, we were unable to demonstrate such an interaction using methods that would have detected binding with a Kd of less than 0.1 mM. PMID- 3032954 TI - Identification of a factor IX/IXa binding protein on the endothelial cell surface. AB - Endothelium provides a specific binding site for Factor IX/IXa which can propagate activation of coagulation by promoting Factor IXa-VIII-mediated activation of Factor X. In this report the endothelial cell Factor IX/IXa binding site has been identified and the coagulant function of the receptor blocked. Studies using [3H]Factor IX derivatized with the photoaffinity labeling agent N succinimidyl-6-(4'-azido-2'-nitrophenylamino)hexanoate (SANPAH) and cultured bovine endothelial cells demonstrated cross-linking to a trypsin-sensitive cell surface protein of Mr approximately equal to 140,000. Immunoprecipitation of metabolically labeled endothelium with Factor IX derivatized with the cleavable cross-linking agent N-succinimidyl(4-azidophenyl)-1,3'-dithiopropionate and antibody to Factor IX demonstrated the endothelial cell origin of the Mr 140,000 cell surface protein. Blockade of the Factor IX/IXa binding protein by covalently linking SANPAH-5-dimethylaminonaphthalene-1-sulfonyl-Glu-Gly-Arg-Factor IXa or SANPAH-Factor IX prevented both specific Factor IXa binding and effective Factor IXa-VIII-mediated activation of Factor X on endothelium. Following extraction of endothelium with detergents, Factor IX/IXa binding activity was solubilized and could be assayed using a polyvinyl chloride plate binding assay. Western blots of cell extracts demonstrated binding of 125I-Factor IX at Mr approximately equal to 140,000 which was blocked by excess Factor IX, but not antisera to Factor VIII, von Willebrand factor, alpha 2-macroglobulin, or epidermal growth factor receptor. These data indicate that endothelium provides a distinct binding site for Factor IX/IXa consisting, at least in part, of a membrane protein which can modulate the coagulant activity of Factor IXa on the cell surface. PMID- 3032956 TI - Nucleotide regulatory protein-mediated activation of phospholipase C in human polymorphonuclear leukocytes is disrupted by phorbol esters. AB - Polymorphonuclear leukocytes (PMNs) activate phospholipase C via a guanine nucleotide regulatory (G) protein. Pretreatment of the PMNs with pertussis toxin (PT) or 4-beta-phorbol 12-myristate 13-acetate (PMA) inhibited chemoattractant induced inositol trisphosphate generation. To determine the loci of inhibition by PT and PMA, G protein-mediated reactions in PMN plasma membranes were examined. Plasma membranes prepared from untreated and PMA-treated PMNs demonstrated equivalent ability of a GTP analogue to suppress high affinity binding of the chemoattractant-N-formyl-methionyl-leucyl-phenylalanine (fMet-Leu-Phe) to its receptor. The rate, but not the extent, of high affinity binding of GTP gamma[35S] to untreated PMN membranes was stimulated up to 2-fold by preincubation with 1 microM fMet-Leu-Phe. The ability of fMet-Leu-Phe to stimulate the rate of GTP gamma S binding was absent in membranes prepared from PT-treated PMNs, but remained intact in membranes from PMA-treated cells. Hydrolysis of phosphatidylinositol 4,5-bisphosphate (PIP2) via phospholipase C could be activated in untreated PMN membranes by either fMet-Leu-Phe plus GTP or GTP gamma S alone at low concentrations of Ca2+ (0.1-1 microM). Membranes prepared from PT-treated PMNs degraded PIP2 upon exposure to GTP gamma S, but not fMet-Leu-Phe plus GTP. In contrast, membranes prepared from phorbol ester-treated PMNs did not hydrolyze PIP2 when incubated with GTP gamma S. Treatment with PT or PMA did not affect the ability of 1 mM Ca2+ to activate PIP2 hydrolysis in PMN membranes, indicating that neither treatment directly inactivated phospholipase C. Therefore, PT appears to block coupling of the chemoattractant receptors to G protein activation, while phorbol esters disrupt coupling of the activated G protein to phospholipase C. The phorbol ester-mediated effect may mimic a negative feedback signal induced by protein kinase C activation by diacylglycerol generated upon activation of phospholipase C. PMID- 3032957 TI - Upper and lower limits of the charge translocation stoichiometry of cytochrome c oxidase. AB - The mechanistic stoichiometry of charge separation coupled to the flow of electrons through cytochrome c oxidase has remained a center of controversy since it was first demonstrated that cytochrome oxidase is an H+ pump. Currently the major dispute is whether the q+/O ratio for this segment is 4 or 6. One cause of the controversy is incomplete coupling between electron flow, electrogenic H+ ejection, and electrophoretic cation uptake, which is usually attributed to finite rates of H+ leakage and/or slippage of the H+ pumps. To minimize the uncertainty which incomplete coupling introduces into estimates of the mechanistic stoichiometry, a new approach (Beavis, A. D., and Lehninger, A. L. (1986) Eur. J. Biochem. 158, 307-314) has been used to determine the upper and lower limits of the mechanistic q+/O translocation stoichiometry of cytochrome oxidase. In this approach, the relationship between the rate of valinomycin dependent K+ uptake, JK, and rate of O2 consumption, JO, is determined as the rates are modulated by two distinct means. When the rates are modulated by the rate of electron flow (i.e. rate of energy supply) the slope of JK versus JO must at all points be less than the mechanistic K+/O ratio. On the other hand, when the rates are modulated by varying the concentration of valinomycin (i.e. the rate of energy utilization) the slope of JK versus JO must at all points be greater than the mechanistic K+/O ratio. The results indicate that the q+/O ratio lies between 4.3 and 5.5. These data are inconsistent with both currently favored stoichiometries, and it is suggested that the true mechanistic stoichiometry of charge separation coupled to electron flow through cytochrome oxidase may be 5 q+/O. PMID- 3032955 TI - Sites of covalent modification in Trg, a sensory transducer of Escherichia coli. AB - The Trg protein mediates chemotactic response of Escherichia coli to the attractants ribose and galactose. Like other transducers, Trg is a transmembrane protein that undergoes post-translational covalent modification. The modifications are hydrolysis (deamidation) of certain glutamine side chains to create glutamate residues and methylation of specific glutamates to form carboxyl methyl esters. Analysis of radiolabeled, tryptic peptides by high performance liquid chromatography and gas-phase sequencing allowed direct identification of the modified residues of Trg. The protein has 5 methyl-accepting residues. Four, at positions 304, 310, 311, and 318, are contained in a 23-residue tryptic peptide ending in lysine. The fifth, at position 500, is within a 25-residue tryptic peptide ending in arginine. At two sites, 311 and 318, glutamines are deamidated to create methyl-accepting glutamates. There is not a required order of modification among the sites. However, there is a substantial preference for methylation on the arginine peptide and, among sites on the lysine peptide, for the middle pair. Comparison of sequences surrounding modified residues identified in this work for Trg and previously for Tsr and Tar suggests a consensus sequence for methyl-accepting sites of Ala/Ser-Xaa-Xaa-Glu-Glu*-Xaa-Ala/OH-Ala-OH/Ala, where OH signifies Ser or Thr and the asterick marks the site of modification. PMID- 3032958 TI - Comparison of the maxicircle (mitochondrial) genomes of Leishmania tarentolae and Trypanosoma brucei at the level of nucleotide sequence. AB - The entire 16.7-kilobase (kb) transcribed region of the Leishmania tarentolae maxicircle was compared to the entire 15-kb transcribed region of the Trypanosoma brucei maxicircle at the nucleotide sequence level by dot matrix analysis and by alignments of individual genes. The L. tarentolae NADH dehydrogenase subunit 1 (ND1) gene was identified in a newly obtained 2.9-kb sequence. All but two regions which flank the cytochrome b gene are highly conserved in both species. One 3.1-kb region in L. tarentolae that contains the cytochrome oxidase subunit III (COIII) gene and several open reading frames corresponds to a 2-kb sequence in T. brucei with limited sequence homology that lacks the COIII gene. Another 0.6-kb region that comprises an unidentified open reading frame (open reading frame 12) in L. tarentolae is substituted by a nonhomologous 0.4-kb open reading frame in T. brucei. A short intergenic region between the ND1 gene and the maxicircle unidentified reading frame 1 gene shows limited sequence homology, and the regions between the ND4 and ND5 genes and between the COI and ND4 genes are not conserved. All of the intergenic regions share G + C richness and a similar pattern of G versus C strand bias. 1.8 kb of the L. tarentolae divergent region (DV) and around 3 kb of the T. brucei DV were also obtained. The T. brucei DV sequences were not homologous to the L. tarentolae DV sequence but were organized in a similar fashion with tandem repeats of varying complexity. PMID- 3032959 TI - Mapping of control elements in the displacement loop region of bovine mitochondrial DNA. AB - The genomes of mammalian mitochondria are duplex DNA circles. The two major transcriptional promoters and the origin of DNA replication for one DNA strand are located in a single region which contains no structural genes and occupies about 6% of the genome. This region is called the displacement loop (D-loop) region since it is often found as a novel triplex structure in which the heavy strand of the genome has been partially replicated. This nascent single-stranded DNA segment remains hybridized to the light strand, displacing the heavy strand of the genome over much of the D-loop region. The promoters and the sites of initiation of D-loop DNA synthesis have been mapped in the human and mouse genomes and may show limited sequence conservation. We have mapped these sites in the bovine mitochondrial genome. Some features are conserved between all three species; however, the promoters and the sites of initiation of D-loop DNA synthesis show no primary sequence homology among all species. This lack of sequence homology is in contrast to the greater than 80% sequence conservation which has been reported in portions of the D-loop region which are located distal to the origin of DNA replication and far from the transcriptional promoters. These results imply that closely related species may have developed different means of controlling mitochondrial gene expression. PMID- 3032960 TI - Sequence specificity of human skin fibroblast collagenase. Evidence for the role of collagen structure in determining the collagenase cleavage site. AB - The sequence specificity of human skin fibroblast collagenase has been investigated by measuring the rate of hydrolysis of 16 synthetic octapeptides covering the P4 through P4' subsites of the substrate. The choice of peptides was patterned after potential collagenase cleavage sites (those containing either the Gly-Leu-Ala or Gly-Ile-Ala sequences) found in types I, II, and III collagens. The initial rate of hydrolysis of the P1-P1' bond of each peptide has been measured by quantitating the concentration of amino groups produced upon cleavage after reaction with fluorescamine. The reactions have been carried out under first-order conditions ([S] much less than KM) and kcat/KM values have been calculated from the initial rates. The amino acids in subsites P3 (Pro, Ala, Leu, or Asn), P2 (Gln, Leu, Hyp, Arg, Asp, or Val), P1' (Ile or Leu), and P4' (Gln, Thr, His, Ala, or Pro) all influence the hydrolysis rates. However, the differences in the relative rates observed for these octapeptides cannot in themselves explain why fibroblast collagenase hydrolyzes only the Gly-Leu and Gly Ile bonds found at the cleavage site of native collagens. This supports the notion that the local structure of collagen is important in determining the location of the mammalian collagenase cleavage site. PMID- 3032961 TI - Cardiac contractile protein phosphatases. Purification of two enzyme forms and their characterization with subunit-specific antibodies. AB - Two forms of protein phosphatase which dephosphorylate cardiac myosin or myosin light chains and the inhibitory subunit of cardiac troponin were purified from bovine cardiac muscle. The enzymes were composed of subunits of Mr = 63,000, 55,000, and 38,000 in a 1:1:1 molar ratio (PT-1) or Mr = 63,000 and 38,000 in a 1:1 molar ratio (PT-2). Native gel electrophoresis and sucrose gradient sedimentation indicated that activity toward all three substrates was due to a single enzyme species. A monoclonal antibody and polyclonal antiserum directed against an Mr = 38,000 protein phosphatase from this tissue specifically reacted with the Mr = 38,000 subunit of PT-1 and PT-2. The specificity of antibodies for the Mr = 38,000 subunit indicated that it was distinct from the other subunits. The Mr = 63,000 subunits of PT-1 and PT-2 were identical based on mobility on sodium dodecyl sulfate gels and one-dimensional peptide maps. Specificity of antiserum against the Mr = 55,000 subunit of PT-1 showed that this subunit was a distinct protein and not derived from the Mr = 63,000 subunit by proteolysis. PT 2 but not PT-1 could interact with antiserum against the Mr = 38,000 catalytic subunit in competitive immunoassays indicating that the presence of the Mr = 55,000 subunit may alter or mask antigenic site(s). Analysis of the enzymatic properties of PT-1 and PT-2 showed that PT-2 had higher activity with myosin, myosin light chains, and phosphorylase while PT-1 had higher activity with troponin. The results indicate that the presence of the Mr = 55,000 subunit may alter the enzymatic properties of the catalytic subunit. PMID- 3032962 TI - Cell- and sequence-specific binding of nuclear proteins to 5'-flanking DNA of the rat growth hormone gene. AB - Stimulation of growth hormone gene transcription in several rat pituitary cell lines (e.g. GC and GH1) is mediated by a thyroid hormone nuclear receptor which is a DNA binding protein. We report that these cell lines contain nuclear proteins which selectively interact with sequences found within the first 236 base pairs of 5'-flanking DNA of the rat growth hormone gene. Sequences found between -104 and -49 base pairs, relative to the transcription initiation (cap) site, bind to nuclear protein(s) which appears to be cell type specific and generate a DNase I-resistant footprint on both strands between -95 and -68. A distinct protein component(s) selectively binds to DNA between -236 and -146 but is not cell type specific. These regions correspond to those found in gene transfer studies to be important in mediating basal expression (-104/+7) and thyroid hormone-regulated expression (-236/-146) of the gene. PMID- 3032963 TI - Two major replicating simian virus 40 chromosome classes. Synchronous replication fork movement is associated with bound large T antigen during elongation. AB - We have analyzed the asynchronous progression of replication forks through the early (E) and late (L) gene sides in bidirectionally replicating SV40 chromosomes during lytic infection. By cutting purified replicating DNA with an appropriate single-site restriction endonuclease and measuring the contour lengths of replicated and unreplicated segments by electron microscopy, the positions of the two replication forks in each elongating intermediate were determined. Our results indicate that there are at least two major classes of replicating SV40 chromosomes which differ in their relative rate of E and L fork movement, the presence or absence of bound SV40 large T antigen during elongation, and the termination region utilized. These two classes also have altered apparent start sites for initiating bidirectional replication, flanking either side of core ori. The largest group (67%) replicated synchronously was associated with T antigen during elongation, appeared to initiate bidirectional elongation at nucleotide 5203 or 41 base pairs (bp) toward the E side of 0/5243, at the junction of T binding site I and ori, and terminated at the typical region centered at 0.5 map units. A second group (24%) replicated asynchronously with the L fork moving 3 times faster than the E fork, was not associated with T antigen during elongation, and terminated at a broad region centered at 0.73 map units. This group appeared to initiate at nucleotide 29 at the junction of the AT-rich region of ori, T binding site I, and the start of the 21-bp repeated transcriptional control sequences. A third group (9%) appeared to initiate at nucleotide 5148 or 95 bp to the E side of 0/5243 and replicated asynchronously preferentially on the E side at early times. However, this group is related to the synchronous class in that it contains bound T antigen and both forks move synchronously past 30% elongation, terminating at the same region. The association of T antigen with synchronous but not asynchronous DNA molecules indicates that T functions in regulating fork movement during elongation. A synchronization role implies that both forks are closely associated with one another in replicating molecules with bound T. Replicating molecules lacking T not only elongated highly asynchronously but preferential fork progression occurred almost exclusively on the L side. The ori region in asynchronous compared to synchronous intermediates was differentially sensitive to BglI digestion, indicating that nuclease digestion can distinguish between different populations of replicating molecules.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3032964 TI - Constitutive function of a positively regulated promoter reveals new sequences essential for activity. AB - A consensus "-10" recognition sequence for RNA polymerase was created at the positively regulated lambda Pre promoter by introducing three single base pair mutations. This altered promoter, Pre*, functions constitutively in vivo and in vitro at high efficiency despite very poor consensus "-35" region sequence homology. We examined the influence of the -35 region sequence information on promoter function by shifting the wild type -35 region +/- 2 base pairs relative to the -10 region consensus sequence and by completely replacing it with alternative DNA sequences. In every case, the altered Pre* promoters retained transcriptional activity although differences in their transcriptional efficiencies were observed. Apparently the Pre* promoter does not require specific -35 region sequences for constitutive promoter activity, although the 35 region sequences can modulate overall promoter strength. In addition, by point mutation analysis we have identified bases immediately upstream of the -10 hexamer which are essential for constitutive function of the Pre* promoter. We propose that these mutants define an extended -10 region at Pre* that compensates for its poor -35 region sequence information by providing critical contacts that stabilize productive RNA polymerase binding. PMID- 3032965 TI - Comparison of the three-dimensional structures of human, yeast, and oat ubiquitin. AB - The crystal structure of human ubiquitin has been solved by x-ray diffraction methods and refined by standard procedures to a conventional crystallographic R factor of 0.176 at 1.8-A resolution (Vijay-Kumar, S., Bugg, C.E., and Cook, W.J. (1987) J. Mol. Biol. 194, 525-538). Crystals of yeast and oat ubiquitin have been grown using human ubiquitin crystals as seeds. Diffraction data for yeast and oat ubiquitin have been collected to a resolution of 1.9 and 1.8 A, respectively. Difference Fourier electron-density maps reveal that the structures of yeast and oat ubiquitin are quite similar to human ubiquitin. All the amino acid changes are clustered in two small patches on one surface of the molecule. This surface is probably not involved in conjugation with proteins destined for ATP-dependent proteolysis. PMID- 3032966 TI - Structure of the Pseudomonas putida alkBAC operon. Identification of transcription and translation products. AB - The structural genes of the Pseudomonas oleovorans alk (alkane utilization) system, which are localized on the alkBAC operon, were cloned as a 16.9-kilobase pair EcoRI fragment. We have measured the length and determined the position of the alkBAC operon on this fragment by electron microscopy of R-loops. Furthermore, the 7.3-kilobase pair long alkBAC operon was analyzed for translation products in Escherichia coli minicells. Using a spectrum of overlapping subclones, six different proteins were identified. Starting from the alkBAC promotor, these polypeptides had molecular masses of 41, 15, 49, 58, 59, and 20 kDa, respectively. The 41-kDa protein was identified as alkane hydroxylase by reaction with a specific antibody. The 15- and 49-kDa peptides are soluble components of the alkane hydroxylase complex. The 58-kDa protein is most likely involved in alkanol dehydrogenase activity. PMID- 3032967 TI - The segment inversion site of herpes simplex virus type 1 adopts a novel DNA structure. AB - The 12-base pair (bp) tandem direct repeat sequences (DR2) at the joint region (a sequence) of herpes simplex virus type 1 (strain F) adopt a new type of DNA conformation under the influence of negative supercoiling. The novel conformation is dependent on the number of the DR2 repeats; the 19 mer (228 bp total) and the 14 mer (168 bp) readily form the alternate structure whereas pentamer, trimer, and dimer repeats show somewhat different properties. S1 and P1 nuclease studies reveal that the new conformation has a major structural aberration at its center and conformational periodicities which are not identical on the complementary strands. Also, the effect of salt and pH, the location of reaction with bromo- and chloroacetaldehyde, the type of sequence (direct repeat) involved, and the nature and extent of supercoil-induced relaxations demonstrate that this structure differs from previously recognized conformations including left-handed Z helices, cruciforms, bent DNA, and slipped structures. We propose the existence of a novel conformation, anisomorphic DNA, with different structures on the complementary strands which elicit a structural aberration at the physical center of the tandem sequences. Since the oligopurine X oligopyrimidine sequence may be inherently inflexible, this supercoil-induced structural change and the physical stress on these inserts in recombinant plasmids tend to deform (crack) the DR2 sequences at their centers. Possible roles for anisomorphic DNA in the functions of this segment of intense biological activity are proposed. PMID- 3032969 TI - A single polypeptide acts both as the beta subunit of prolyl 4-hydroxylase and as a protein disulfide-isomerase. AB - A single polypeptide is shown to act both as the beta subunit of the proline hydroxylase (EC 1.14.11.2) and as a protein disulfide-isomerase (EC 5.3.4.1). When isolated from chick embryos or rat liver, the beta subunit of prolyl 4 hydroxylase and the enzyme protein disulfide-isomerase have identical molecular weights and peptide maps as produced by digestion with Staphylococcus aureus V8 protease. The apparent molecular weights of both proteins isolated from human placental tissue are slightly higher, and the human beta subunit and one of its peptides have molecular weights about Mr 500 higher than the protein disulfide isomerase and its corresponding peptide. Experiments with polyclonal and monoclonal antibodies also suggest a structural identity between the two proteins. The beta subunit isolated from the prolyl 4-hydroxylase tetramer has protein disulfide-isomerase activity similar to protein disulfide-isomerase itself, and even the beta subunit when present in the prolyl 4-hydroxylase tetramer has one-half of this activity. PMID- 3032968 TI - The maximal velocity and the calcium affinity of the red cell calcium pump may be regulated independently. AB - The kinetics of active Ca2+ transport in inside-out red cell membrane vesicles and the Ca2+-ATPase activity of the purified Ca2+ pump were studied and the effects of calmodulin, acidic phospholipids, and controlled trypsinization were compared. In the presence of calmodulin the maximal rate and the apparent affinity of the pump for Ca2+ were greatly increased in both preparations. The lowest value of Km(Ca) was between 0.5 and 0.7 microM depending on the concentration of calmodulin and on the enzyme preparation. Positive cooperativity for Ca2+ activation with a Hill coefficient of 1.6-1.7 was observed in all cases. When acidic phospholipids (phosphatidylinositol 4-phosphate was routinely used) were added to the inside-out vesicles or to the purified enzyme, maximal transport rates equal to those obtained with calmodulin were measured but the Km(Ca) decreased to 0.25 microM and the positive cooperativity disappeared (the Hill coefficient approached 1). Highly active, calmodulin-independent proteolytic fragments of molecular mass of 81 and 76 kDa were produced with controlled trypsinization. When the trypsin treatment was directed to obtain primarily the 81-kDa fragment, the preparation showed characteristics similar to those of the intact Ca2+ pump in the presence of calmodulin; that is, the same Vmax was obtained, the Km(Ca2+) was 0.5-0.6 microM, and the Hill coefficient was about 1.6. Addition of phosphatidylinositol 4-phosphate or allowing further proteolysis to produce the 76-kDa fragment, shifted the Km(Ca) to 0.25 and reduced the Hill coefficient to 1, without changes in the maximal rate. Based on these results it is suggested that the maximal velocity and the Ca2+ affinity on the erythrocyte Ca2+ pump may be regulated independently and that independent polypeptide regions of the enzyme are involved in the regulations. PMID- 3032970 TI - Activation of the respiratory burst oxidase in a fully soluble system from human neutrophils. AB - The O2(-)-forming respiratory burst oxidase is present in a dormant state in a fully soluble system containing both cytosol and a deoxycholate extract of membranes from resting human neutrophils. Sodium dodecyl sulfate at low concentrations converts this soluble dormant oxidase into its catalytically active form. The Vmax for the activated oxidase was 2.1 mumol of O2-/min/mg of membrane protein. Michaelis constants for NADPH and NADH (38 microM and 1.7 mM, respectively) were similar to those measured previously in other systems. Oxidase activity was not detected after sodium dodecyl sulfate treatment of systems containing solubilized neutrophil membranes obtained from patients with X-linked chronic granulomatous disease. These results suggest that the deoxycholate extract contains both the resting oxidase and those membrane-associated components needed for its activation, all in functioning states. PMID- 3032971 TI - Regulation by phosphatidylinositol of rat pituitary plasma membrane and endoplasmic reticulum phosphatidylinositol synthase activities. A mechanism for activation of phosphoinositide resynthesis during cell stimulation. AB - The mechanism of signal transduction used by a large number of extracellular regulatory molecules involves hydrolysis and resynthesis of phosphoinositides. We recently demonstrated that during stimulation by thyrotropin-releasing hormone of rat pituitary (GH3) cells phosphatidylinositol (PtdIns) resynthesis occurs within the plasma membrane as well as the endoplasmic reticulum (Imai, A., and Gershengorn, M. C. (1987) Nature, 325, 726-728). In this report, we have studied regulation of PtdIns synthase (CDP-diglyceride-inositol phosphatidyltransferase, EC 2.7.8.11) activities associated with plasma membranes and endoplasmic reticulum isolated from GH3 cells. Exogenously added PtdIns noncompetitively inhibited membrane-associated and solubilized PtdIns synthase activities by up to 84 to 91%; half-maximal inhibition occurred between 0.03 and 0.1 mM PtdIns. Similar inhibition of PtdIns synthase activities were observed when PtdIns content of both membrane fractions was increased in vivo in intact GH3 cells prior to assay in vitro. These findings demonstrate that PtdIns synthase activities associated with plasma membrane and endoplasmic reticulum fractions isolated from GH3 cells are inhibited by the product, PtdIns. Because PtdIns levels decrease and PtdIns resynthesis is activated in both membrane fractions during stimulation of GH3 cells by thyrotropin-releasing hormone, it seems likely that activation of PtdIns synthase(s) during cell stimulation occurs by release of this enzyme(s) from inhibition by its product. PMID- 3032973 TI - EPR and electron nuclear double resonance investigation of oxidized hydrogenase II (uptake) from Clostridium pasteurianum W5. Effects of carbon monoxide binding. AB - Two different hydrogenases have been isolated from Clostridium pasteurianum W5. Hydrogenase II (uptake) is active in H2 oxidation while hydrogenase I (bidirectional) is active both in H2 oxidation and evolution. Previous EPR and electron nuclear double resonance (ENDOR) studies of oxidized hydrogenase I have now been complemented by analogous studies on oxidized 57Fe-enriched hydrogenase II and its CO derivative (using 12CO and 13CO). Binding of CO greatly changes the EPR spectrum of oxidized hydrogenase II, and use of 13CO leads to resolved hyperfine splitting from interaction with a single 13CO molecule (AC approximately 34 MHz). This coupling is over 50% larger than that seen for hydrogenase I. 57Fe ENDOR disclosed two types of iron site in both oxidized hydrogenase II and its CO derivative. Combination of EPR, ENDOR, and Mossbauer results shows that site 1 has AFe1 = 18 MHz shifting to approximately 30 MHz upon CO binding and consisting of two Fe atoms and site 2 has A2 approximately 7 MHz shifting to approximately 10 MHz and containing a single Fe. These results are very similar to those seen for hydrogenase I, which indicates that a structurally similar 3Fe cluster, believed to be the catalytically active site, is present in both. Proton ENDOR shows a solvent exchangeable resonance only in the CO derivative of hydrogenase II. This indicates a structural difference between hydrogenases I and II that is brought out by CO binding. No evidence of 14N coordination to the cluster is seen for either enzyme. PMID- 3032972 TI - Purification and some characteristics of nitrous oxide reductase from Paracoccus denitrificans. AB - Nitrous oxide reductase from the denitrifying bacterium Paracoccus denitrificans has been purified very nearly to homogeneity by an anaerobic procedure that results in a product with high specific activity. The enzyme is a dimer of about Mr 144,000 composed of two subunits of apparently equal Mr and contains 4 mol of Cu per mol of subunit. The isoelectric point is 4.3; specific activity at 25 degrees C, pH 7.1, is 122 mumol X min-1 X mg of protein-1; and Km is about 7 microM N2O under the same conditions. The N2O- and O2-oxidized forms of the enzyme had principal absorption bands at 550 and 820 nm; the dithionite-reduced form, at 650 nm. The extinction coefficient at 550 nm for the oxidized enzyme is about 5300 (M subunit)-1 X cm-1. Ferricyanide-oxidized enzyme and enzyme exposed to O2 for a couple of days at 4 degrees C exhibited additional bands at 480, 620, and 780 nm and had very low specific activities. Cu-EPR signals were observed with oxidized and reduced forms of the enzyme with g perpendicular values at 2.042 and 2.055, respectively. The O2-oxidized enzyme had g parallel and A parallel values of about 2.244 and 35 gauss, respectively, based on the observation of four hyperfine lines in the g parallel region. The enzyme may therefore contain at least one Cu atom approximating the "Type 1" class. Spin counts against Cu-EDTA standards suggest that 20-30% of the enzyme-bound Cu is EPR detectable in the O2-oxidized enzyme and 7-15% in the enzyme as prepared and in the reduced enzyme. Much of the Cu thus appears to be EPR silent. Nitrous oxide reductase was observed to undergo turnover-dependent inactivation, and nitrite and fluoride among other anions were found to accelerate this process. In a number of characteristics, the enzyme resembles nitrous oxide reductase recently purified from Pseudomonas perfectomarina and Rhodopseudomonas sphaeroides, particularly the former. Some differences appear related to whether or not purification is carried out entirely under anaerobic conditions. PMID- 3032974 TI - Polymyxin B is an inhibitor of insulin-induced hypoglycemia in the whole animal model. Studies on the mode of inhibitory action. AB - The cyclic decapeptide, polymyxin B (PMXB), was found to inhibit hypoglycemia in mice receiving exogenous insulin (Amir, S., and Shechter, Y. (1985) Eur. J. Pharmacol. 110, 283-285). In this study, we have extended this observation to rats. Insulin-dependent hypoglycemia in rats is efficiently blocked at a 12:1 molar ratio of PMXB to insulin. This effect is highly specific, as it could not be mimicked by a variety of antibiotics or positively charged substances. Chemical modifications of PMXB have revealed that the ring structure, rather than the tail structure, is important for anti-insulin-like activity. Colistin A, which differs from PMXB by one conservative amino acid substitution in the ring structure, is devoid of this activity. Polymyxin B does not interact with insulin, nor does it alter the rate of insulin absorption and/or degradation, or the ability of insulin to bind to target tissues. This peptide inhibits hypoglycemia by blocking insulin-dependent activation of the hexose transport mechanism, as deduced by in vitro studies. The effect of insulin in stimulating hexose uptake (and subsequent glucose metabolism) in both isolated muscle tissue and adipocytes is blocked with little or no effect on the basal activities of these processes. Colistin A has no significant inhibiting effect. Other insulin dependent activities, such as inhibition of lipolysis in adipocytes or synthesis of DNA in muscle cells, are not inhibited. It is concluded that PMXB inhibits, in a highly specific manner, the action of insulin in stimulating hexose transport and subsequent glucose metabolism, both in vitro and in the whole animal model. PMID- 3032975 TI - Expression of completely gamma-carboxylated recombinant human prothrombin. AB - Human prothrombin cDNA has been expressed in mammalian cells to yield biologically active, fully gamma-carboxylated prothrombin. A 2.0-kilobase cDNA encoding full-length prothrombin was isolated from a human fetal liver library using a cDNA fragment recovered from a lambda gt11 human hepatoma expression library. Prothrombin cDNA was cloned into a mammalian expression vector and transfected into Chinese hamster ovary cells. Selection for expression of dihydrofolate reductase yielded cell lines secreting up to 0.55 microgram/ml of prothrombin. Recombinant prothrombin synthesized in the presence of vitamin K was quantitatively recovered from tissue culture medium by affinity chromatography using conformation-specific antibodies directed against the metal-stabilized, gamma-carboxylated conformer. The purified material migrated as a single band on denaturing polyacrylamide gels with an electrophoretic mobility equivalent to that of plasma-derived human prothrombin. Automated Edman degradation of recombinant prothrombin revealed a single amino-terminal sequence identical to that of plasma-derived prothrombin. Recombinant and plasma-derived prothrombin interacted similarly with antibodies specific for total prothrombin, abnormal des gamma-carboxyprothrombin, and two metal-stabilized conformers of prothrombin. Recombinant prothrombin exhibited a specific coagulant activity equivalent to that of plasma-derived prothrombin. The gamma-carboxyglutamic acid analysis of recombinant prothrombin demonstrated 9.9 +/- 0.4 mol of gamma-carboxyglutamic acid/mol of prothrombin. These results represent the first description of the expression of a recombinant vitamin K-dependent protein in which all of the expressed protein is gamma-carboxylated. PMID- 3032976 TI - Nucleotide sequence and organization of the rat heme oxygenase gene. AB - Heme oxygenase, an essential enzyme of heme catabolism, is inducible by its substrate heme, by heavy metals, and by various other substances. To study the molecular mechanisms of the induction of heme oxygenase, we isolated the heme oxygenase gene from a rat genomic DNA library using cloned cDNA as hybridization probes and determined its complete nucleotide sequence. The gene is composed of 6830 nucleotides, and is organized in four introns and five exons. The transcription initiation site was identified by S1 nuclease mapping analysis. Using HeLa cell lysate, we confirmed that the transcription of cloned heme oxygenase gene is initiated accurately at the assigned initiation site. In the 5' flanking region of the heme oxygenase gene, we found several potential binding sites for different transcription factors: a transcription factor Sp1, a positive regulator for the control of amino acid synthesis (GCN4), a heat shock transcription factor, and a metal-dependent transcription factor. Furthermore, the intron 1 contains the sequence that shows about 65% homology to that of the neuronal identifier sequence, a possible enhancer element. PMID- 3032977 TI - Two Na,K-ATPase isoenzymes in canine cardiac myocytes. Molecular basis of inotropic and toxic effects of digitalis. AB - Canine cardiac myocytes contain two distinct molecular forms of the Na,K-ATPase catalytic subunit. They are resolved by gel electrophoresis and identified using immunological techniques. The apparent molecular weights of the catalytic subunits are 95,000 (alpha) and 98,000 (alpha +). As judged by [3H]ouabain binding measurements and Na,K-ATPase assays, the two forms are active and differ by a factor of 150 in their respective affinity for digitalis (ouabain and digitoxigenin). The dissociation constant of the high affinity form (alpha +) is KD, 2 nM, and that of the low affinity molecular form (alpha) is KD, 300 nM. According to both enzymatic and binding assays, up to 70% of maximum inhibition is caused by occupation of the high affinity sites (alpha +). Inasmuch as the pharmacological and toxic concentrations of digitalis in dog are 1 and 200 nM, respectively, and as maximum inhibition of Na+ pump in vivo should not exceed 80% to avoid toxicity (Akera, T. and Brody, T. (1982) Annu. Rev. Physiol. 44, 375 388), it appears that the high affinity molecular form (alpha +) is the pharmacological receptor exclusively related to positive inotropy, whereas the low affinity form (alpha) is mainly associated with toxicity. PMID- 3032978 TI - Transcription of Rhizobium meliloti nodulation genes. Identification of a nodD transcription initiation site in vitro and in vivo. AB - Nodulation genes in Rhizobium are required for invasion of the host plant. The nodABC operon is induced by plant activator molecules; this activation requires the gene product of the constitutively expressed nodD locus, which is transcribed divergently from nodABC. We are employing in vitro transcription to elucidate the molecular mechanism of nod gene activation. We used a micropurification technique to obtain RNA polymerase from Rhizobium meliloti, and here demonstrate that it initiated and terminated accurately at the Escherichia coli trp promoter-leader region. E. coli RNA polymerase, however, apparently fails to recognize R. meliloti promoters. We used the R. meliloti RNA polymerase in a minimal transcription system to attempt to localize the divergent start sites for nodD and nodABC. Transcript sizing and fingerprinting, together with synchronized single-round transcription experiments permit us to designate an in vitro transcription initiation site for nodD. Primer extension analysis of in vivo mRNA demonstrates that the initiation site which is utilized in vitro is the same site used in vivo. While nodABC is not transcribed in our minimal in vitro transcription system, this system should prove useful for the study of factors in induced cells which promote expression of this inducible promoter. PMID- 3032979 TI - Helicase action of dnaB protein during replication from the Escherichia coli chromosomal origin in vitro. AB - Initiation of bidirectional replication from the origin of the Escherichia coli chromosome (oriC) proceeds through stages in which the components of the two replication forks are assembled. From a complex containing proteins dnaA, dnaB, and dnaC bound at oriC, the dnaB helicase moves in both directions to unwind the duplex. In the absence of replication, this unwinding generates a bubble at oriC coated by single strand binding protein. Addition of gyrase allows unwinding to proceed extensively in both directions from oriC at 60 base pairs/s/fork at 37 degrees C. This rate is sharply dependent on temperature and also stimulated by both primase and DNA polymerase III holoenzyme, even in the absence of DNA synthesis. Primer and DNA synthesis are efficient when coupled to template unwinding. DNA synthesis proceeds bidirectionally from oriC at a rate limited by unwinding. With extensive unwinding preceding DNA synthesis, initiations are not limited to oriC. PMID- 3032980 TI - Regulation of estrogen biosynthesis in human adipose stromal cells. Effects of dibutyryl cyclic AMP, epidermal growth factor, and phorbol esters on the synthesis of aromatase cytochrome P-450. AB - Using human adipose stromal cells in monolayer culture as a model system for study of the regulation of aromatase activity, as well as polyclonal antibodies raised in this laboratory against aromatase cytochrome P-450 (cytochrome P 450AROM), it was found that the rate of synthesis of cytochrome P-450AROM was stimulated by dibutyryl cyclic AMP. This stimulation was attenuated by epidermal growth factor and was potentiated by phorbol esters. These changes in cytochrome P-450AROM synthesis were associated with comparable changes in the levels of translatable cytochrome P-450AROM mRNA, as well as with changes in the activity of aromatase of these cells. By contrast, there was little change in the synthesis of the reductase component of the aromatase enzyme complex in response to these factors. The increase in mRNA was blocked by cycloheximide, indicative of a requirement for protein synthesis in mediating this inductive response. It is concluded that aromatase activity is regulated primarily by changes in the level of mRNA encoding cytochrome P-450AROM, and that such changes are likely a reflection of changes in the rate of transcription of the gene encoding this enzyme. Increases in the levels of cytochrome P-450AROM mRNA are apparently mediated by a regulatory protein(s), similar to that found for other steroidogenic forms of cytochrome P-450. PMID- 3032981 TI - Characterization of lipocortin I and an immunologically unrelated 33-kDa protein as epidermal growth factor receptor/kinase substrates and phospholipase A2 inhibitors. AB - Reversible calcium-dependent association with a particulate fraction from human placenta was used as the first step in the purification of substrates for the epidermal growth factor-stimulated protein kinase. A protein with apparent Mr of 35,000 was purified to homogeneity, and the sequence was determined for approximately one-fourth of the protein. These residues could be aligned exactly with the previously published sequence of lipocortin I derived from the cDNA from a human lymphoma. Two other proteins that appear to be formed by proteolytic removal of 12 or 26 of the amino acids from the NH2 terminus of the protein also were isolated. Placental lipocortin I was phosphorylated in Tyr-21 in an epidermal growth factor-dependent manner by the kinase activity in a particulate fraction from A431 cells; half-maximal phosphorylation occurred at 50 nM lipocortin I. Lipocortin I phosphorylated on Tyr-21 was approximately 10-fold more sensitive to tryptic cleavage at Lys-26 than was the native protein. Placental lipocortin I and its two truncated forms were potent inhibitors of pancreatic phospholipase A2 activity. Another 33-kDa protein that was not related immunologically to lipocortin I or lipocortin II (calpactin I) also was purified from the EGTA extract of placenta. The unidentified protein inhibited phospholipase A2 but was not a substrate for the epidermal growth factor stimulated kinase. The mechanism by which these proteins inhibit phospholipase A2 activity was investigated. Attempts to detect direct interaction between these proteins and the enzyme were unsuccessful. However, both the unidentified protein, lipocortin I, and 32P-labeled lipocortin I bound in a Ca2+-dependent manner to the [3H]oleic acid-labeled Escherichia coli membranes used as substrate in the phospholipase A2 assay. Heparin, which is known to block lipocortin I inhibition of phospholipase A2, also blocked binding of lipocortin I to E. coli membranes. The results of these and other experiments raise the possibility that placental lipocortin I inhibits phospholipase A2 activity in this assay by coating the phospholipid and thereby blocking interaction of enzyme and substrate. PMID- 3032982 TI - Femoral neuropathy secondary to pressurized cement in total hip replacement: treatment by decompression and neurolysis. Report of a case. PMID- 3032983 TI - Yeast carboxypeptidase Y can be translocated and glycosylated without its amino terminal signal sequence. AB - We have constructed a series of mutations in the signal sequence of the yeast vacuolar protein carboxypeptidase Y (CPY), and have used pulse-chase radiolabeling and immunoprecipitation to examine the in vivo effects of these mutations on the entry of the mutant CPY proteins into the secretory pathway. We find that introduction of a negatively charged residue, aspartate, into the hydrophobic core of the signal sequence has no apparent effect on signal sequence function. In contrast, internal in-frame deletions within the signal sequence cause CPY to be synthesized as unglycosylated precursors. These are slowly and inefficiently converted to glycosylated precursors that are indistinguishable from the glycosylated forms produced from the wild-type gene. These precursors are converted to active CPY in a PEP4-dependent manner, indicating that they are correctly localized to the vacuole. Surprisingly, a deletion mutation that removes the entire CPY signal sequence has a similar effect: unglycosylated precursor accumulates in cells carrying this mutant gene, and greater than 10% of it is posttranslationally glycosylated. Thus, the amino-terminal signal sequence of CPY, while important for translocation efficiency, is not absolutely required for the translocation of this protein. PMID- 3032984 TI - Regulation by aldosterone of Na+,K+-ATPase mRNAs, protein synthesis, and sodium transport in cultured kidney cells. AB - Transepithelial Na+ reabsorption across tight epithelia is regulated by aldosterone. Mineralocorticoids modulate the expression of a number of proteins. Na+,K+-ATPase has been identified as an aldosterone-induced protein (Geering, K., M. Girardet, C. Bron, J. P. Kraehenbuhl, and B. C. Rossier, 1982, J. Biol. Chem., 257:10338-10343). Using A6 cells (kidney of Xenopus laevis) grown on filters we demonstrated by Northern blot analysis that the induction of Na+,K+-ATPase was mainly mediated by a two- to fourfold accumulation of both alpha- and beta subunit mRNAs. The specific competitor spironolactone decreased basal Na+ transport, Na+,K+-ATPase mRNA, and the relative rate of protein biosynthesis, and it blocked the response to aldosterone. Cycloheximide inhibited the aldosterone dependent sodium transport but did not significantly affect the cytoplasmic accumulation of Na+,K+-ATPase mRNA induced by aldosterone. PMID- 3032985 TI - Localization of Na+,K+-ATPase alpha-subunit to the sinusoidal and lateral but not canalicular membranes of rat hepatocytes. AB - Controversy has recently developed over the surface distribution of Na+,K+-ATPase in hepatic parenchymal cells. We have reexamined this issue using several independent techniques. A monoclonal antibody specific for the endodomain of alpha-subunit was used to examine Na+,K+-ATPase distribution at the light and electron microscope levels. When cryostat sections of rat liver were incubated with the monoclonal antibody, followed by either rhodamine or horseradish peroxidase-conjugated goat anti-mouse secondary, fluorescent staining or horseradish peroxidase reaction product was observed at the basolateral surfaces of hepatocytes from the space of Disse to the tight junctions bordering bile canaliculi. No labeling of the canalicular plasma membrane was detected. In contrast, when hepatocytes were dissociated by collagenase digestion, Na+,K+ ATPase alpha-subunit was localized to the entire plasma membrane. Na+,K+-ATPase was quantitated in isolated rat liver plasma membrane fractions by Western blots using a polyclonal antibody against Na+,K+-ATPase alpha-subunit. Plasma membranes from the basolateral domain of hepatocytes possessed essentially all of the cell's estimated Na+,K+-ATPase catalytic activity and contained a 96-kD alpha subunit band. Canalicular plasma membrane fractions, defined by their enrichment in alkaline phosphatase, 5' nucleotidase, gamma-glutamyl transferase, and leucine aminopeptidase had no detectable Na+,K+-ATPase activity and no alpha-subunit band could be detected in Western blots of these fractions. We conclude that Na+,K+ ATPase is limited to the sinusoidal and lateral domains of hepatocyte plasma membrane in intact liver. This basolateral distribution is consistent with its topology in other ion-transporting epithelia. PMID- 3032986 TI - Sorting of endocytosed transferrin and asialoglycoprotein occurs immediately after internalization in HepG2 cells. AB - After receptor-mediated uptake, asialoglycoproteins are routed to lysosomes, while transferrin is returned to the medium as apotransferrin. This sorting process was analyzed using 3,3'-diaminobenzidine (DAB) cytochemistry, followed by Percoll density gradient cell fractionation. A conjugate of asialoorosomucoid (ASOR) and horseradish peroxidase (HRP) was used as a ligand for the asialoglycoprotein receptor. Cells were incubated at 0 degree C in the presence of both 131I-transferrin and 125I-ASOR/HRP. Endocytosis of prebound 125I-ASOR/HRP and 131I-transferrin was monitored by cell fractionation on Percoll density gradients. Incubation of the cell homogenate in the presence of DAB and H2O2 before cell fractionation gave rise to a density shift of 125I-ASOR/HRP containing vesicles due to HRP-catalyzed DAB polymerization. An identical change in density for 125I-transferrin and 125I-ASOR/HRP, induced by DAB cytochemistry, is taken as evidence for the concomitant presence of both ligands in the same compartment. At 37 degrees C, sorting of the two ligands occurred with a half time of approximately 2 min, and was nearly completed within 10 min. The 125I ASOR/HRP-induced shift of 131I-transferrin was completely dependent on the receptor-mediated uptake of 125I-ASOR/HRP in the same compartment. In the presence of a weak base (0.3 mM primaquine), the recycling of transferrin receptors was blocked. The cell surface transferrin receptor population was decreased within 6 min to 15% of its original size. DAB cytochemistry showed that sorting between endocytosed 131I-transferrin and 125I-ASOR/HRP was also blocked in the presence of primaquine. These results indicate that transferrin and asialoglycoprotein are taken up via the same compartments and that segregation of the transferrin-receptor complex and asialoglycoprotein occurs very efficiently soon after uptake. PMID- 3032988 TI - Quantitation of an alpha subunit splicing intermediate: evidence for transcriptional activation in the control of acetylcholine receptor expression in denervated chick skeletal muscle. AB - We have investigated the mechanisms responsible for the increase in acetylcholine receptor subunit mRNAs during the induction of denervation supersensitivity in skeletal muscle. Using a cRNA probe specific for exon 7 (224 nucleotides; with flanking intron sequences of 105 nucleotides on the 3' end, and of 70 nucleotides on the 5' end) of the alpha subunit of the chicken muscle acetylcholine receptor gene, we were able to quantitate the concentration of mature alpha subunit mRNA and its precursor. In 3-wk-old chicks, the concentration of alpha subunit message in leg muscle was found to be 4.0 attomoles per microgram total RNA, and to increase 40-fold within 1 wk after section of the sciatic nerve. The molar ratio of precursor/mature mRNA, which was approximately 0.023 in innervated as well as denervated muscle, transiently rose to 0.047 at the beginning of the second postoperative day when mature message content increased 20-fold; the rise in precursor level preceded the increase in mature message content. These findings suggest that an accelerated rate of transcription of the message coding for the alpha subunit causes increased message content and the stimulation of receptor synthesis characteristic of denervated muscle. PMID- 3032989 TI - Release of angiotensin I-converting enzyme by endothelial cells in vitro. AB - Bovine fetal aortic endothelial cells cultured in serum-containing medium accumulate angiotensin I-converting enzyme (ACE) activity and also release it into the culture medium. Following subcultivation of a confluent culture using trypsin-EDTA, cellular ACE activity falls 50% within 8 h, but no ACE activity is detected in the medium, suggesting intracellular loss of the enzyme activity. ACE activity reappears in both the cell lysate and culture medium after the culture becomes confluent. The rate of accumulation of ACE activity released into the medium is always greater than that for cellular activity. For example, 21 days following subcultivation 80-85% of the total culture activity is detected in the medium. Both cellular and medium-associated ACE decrease proportionately as the culture progresses through its in vitro lifespan. PMID- 3032987 TI - Myosin light chain kinase and myosin light chain phosphatase from Dictyostelium: effects of reversible phosphorylation on myosin structure and function. AB - We have partially purified myosin light chain kinase (MLCK) and myosin light chain phosphatase (MLCP) from Dictyostelium discoideum. MLCK was purified 4,700 fold with a yield of approximately 1 mg from 350 g of cells. The enzyme is very acidic as suggested by its tight binding to DEAE. Dictyostelium MLCK has an apparent native molecular mass on HPLC G3000SW of approximately 30,000 D. Mg2+ is required for enzyme activity. Ca2+ inhibits activity and this inhibition is not relieved by calmodulin. cAMP or cGMP have no effect on enzyme activity. Dictyostelium MLCK is very specific for the 18,000-D light chain of Dictyostelium myosin and does not phosphorylate the light chain of several other myosins tested. Myosin purified from log-phase amebas of Dictyostelium has approximately 0.3 mol Pi/mol 18,000-D light chain as assayed by glycerol-urea gel electrophoresis. Dictyostelium MLCK can phosphorylate this myosin to a stoichiometry approaching 1 mol Pi/mol 18,000-D light chain. MLCP, which was partially purified, selectively removes phosphate from the 18,000-D light chain but not from the heavy chain of Dictyostelium myosin. Phosphatase-treated Dictyostelium myosin has less than or equal to 0.01 mol Pi/mol 18,000-D light chain. Phosphatase-treated myosin could be rephosphorylated to greater than or equal to 0.96 mol Pi/mol 18,000-D light chain by incubation with MLCK and ATP. We found myosin thick filament assembly to be independent of the extent of 18,000-D light-chain phosphorylation when measured as a function of ionic strength. However, actin-activated Mg2+-ATPase activity of Dictyostelium myosin was found to be directly related to the extent of phosphorylation of the 18,000-D light chain. MLCK-treated myosin moved in an in vitro motility assay (Sheetz, M. P., and J. A. Spudich, 1983, Nature (Lond.), 305:31-35) at approximately 1.4 micron/s whereas phosphatase-treated myosin moved only slowly or not at all. The effects of phosphatase treatment on the movement were fully reversed by subsequent treatment with MLCK. PMID- 3032990 TI - High and low affinity binding sites for basic fibroblast growth factor on cultured cells: absence of a role for low affinity binding in the stimulation of plasminogen activator production by bovine capillary endothelial cells. AB - Scatchard analysis of binding of 125I-basic fibroblast growth factor (FGF) to baby hamster kidney (BHK) cells revealed the presence of two binding sites: a high affinity site with KD of 20 pM and 80,000 sites per cell and a low affinity site with KD of about 2 nM and 600,000 sites per cell. The binding to the two sites could be separated by first washing the cells with 2 M NaCl at pH 7.5 which released the low affinity binding and then extracting the cells with 0.5% Triton X-100 to recover the 125I-basic FGF bound to high affinity sites. The binding to the high affinity site was acid sensitive, suggesting that it represented binding to the receptor. Binding to the low affinity site could be competed strongly by heparin and less strongly by heparan sulfate but not by chondroitin sulfate, dermatan sulfate, or keratan sulfate. Treatment of BHK cells with heparinase abolished 62% of the low affinity binding, suggesting that the low affinity binding represented binding to cell-associated, heparin-like molecules. A variety of other cell types, including bovine capillary endothelial (BCE) cells, also demonstrated both low and high affinity binding sites. To test whether the low affinity binding might play a role in the basic FGF stimulation of plasminogen activator (PA) production by BCE cells, heparin was added to BCE cultures at concentrations which totally blocked binding of 125I-basic FGF to the low affinity sites. Addition of the heparin did not diminish the increased PA production induced by basic FGF. This suggests that the low affinity binding has no direct role in the stimulation of PA production in BCE cells. PMID- 3032991 TI - Heparin modulation of the neurotropic effects of acidic and basic fibroblast growth factors and nerve growth factor on PC12 cells. AB - Nerve growth factor (NGF) and acidic or basic fibroblast growth factor (aFGF and bFGF, respectively) induce neurite outgrowth from the rat pheochromocytoma cell line, PC12. The neurites induced by these three factors are stable for up to a month in cell culture in the continued presence of any of the above growth factors. bFGF (ED50 = 30 pg/ml) is 800 fold more potent in stimulating neurite outgrowth than aFGF (ED50 = 25 ng/ml) and 260 fold more potent than NGF (ED50 = 8 ng/ml). While the neurotropic activities of aFGF and NGF are potentiated by heparin, that of bFGF is both partially inhibited or stimulated, depending upon the concentration of bFGF. Radioreceptor binding experiments show that aFGF and bFGF bind to a common binding site on the PC12 cell surface. Affinity labeling studies demonstrate a single receptor with an apparent molecular weight of 145,000 daltons, which corresponds to the high molecular weight receptor identified in BHK-21 cells. NGF does not appear to compete with aFGF or bFGF for binding to the receptor. Heparin blocked the binding of bFGF to the receptor but had only a small inhibitory effect on the binding of aFGF to the receptor. Thus, it appears that heparin inhibition of the neurotropic effects of bFGF occurs, at least in part, by impairing the interaction of bFGF with the receptor, while having little effect on that of aFGF. The stimulatory effects of heparin on the neurotropic activity of aFGF, bFGF, and NGF may occur through a site not associated with the respective cellular receptor for the growth factors. PMID- 3032992 TI - Effect of reduced endocytosis induced by hypotonic shock and potassium depletion on the infection of Hep 2 cells by picornaviruses. AB - Potassium depletion after a brief exposure of the cells to hypotonic medium was used to inhibit endocytosis from coated pits in Hep 2 cells. After such treatment the endocytic uptake of transferrin was arrested, and electron microscopy revealed that virtually no coated pits were present at the cell surface, while smooth (uncoated) pits were abundant. Under the same conditions the cells were strongly protected against poliovirus, while the cytopathogenic effect of human rhinovirus type 2, HRV 2, was increased. The cytopathogenic effect of encephalomyocarditis (EMC) virus was only slightly affected. Potassium depletion without hypotonic shock reduced the endocytic uptake of transferrin 2-3-fold and the number of coated pits at the cell surface about 3-fold. Furthermore, the cells were not protected against poliovirus after such treatment. The data indicate that the productive uptake of poliovirus occurs by receptor-mediated endocytosis from coated pits, while the productive uptake of the other two picornaviruses may occur by another endocytic pathway. In order to efficiently arrest endocytosis from coated pits in these cells, hypotonic shock seems to be a critical component of the potassium depletion protocol. PMID- 3032994 TI - Magnesium deprivation inhibits the expression of differentiation-related phenotypes in human promyelocytic leukemia HL-60 cells. AB - The role of magnesium ions in the differentiation of human promyelocytic leukemia HL-60 cells was investigated. When HL-60 extracellular magnesium was deficient (less than 0.01 mM), the total intracellular magnesium content and [3H] leucine incorporation rates decreased to 61 and 28%, respectively, on day 3. When the cells were treated with various inducers (100 nM 1 alpha, 25 dihydroxyitamine D3 (1,25(OH)2D3), 100 nM beta-all-trans retinoic acid (RA), 20 nM 12-o-tetradecanoyl phorbol-13-acetate (TPA), 1.25% dimethylsulfoxide (DMSO) and 30 nM aclacinomycin (AcM] in magnesium-deficient medium, the expression of differentiation-related phenotypes (nitroblue tetrazolium (NBT) reducing ability, nonspecific esterase (NSE) activity and monoclonal antibody, OKM1 binding activity) was almost completely inhibited. After a 2-day treatment with 100 nM 1,25(OH)2D3 in magnesium-deficient medium, the expression of differentiation-related phenotypes was restored by further incubation in the absence of inducer in standard magnesium medium (0.4 mM). These results suggested that magnesium deprivation inhibited the expression of HL-60 differentiation-related phenotypes but not their commitment to differentiation. These phenotypes were expressed without inducer in standard magnesium medium after a 2-day simultaneous treatment with 1,25(OH)2D3 and cyclohexamide (protein synthesis inhibitor) in magnesium deficient medium, but not after simultaneous pretreatment with 1,25(OH)2D3 and alpha-amanitin (RNA synthesis inhibitor). Thus, it was suggested that the magnesium-requiring step in HL-60 cell differentiation is in protein but not mRNA synthesis. This conclusion is supported by the findings that changes in c-myc and c-fms mRNA levels in HL-60 cells treated with 100 nM 1,25(OH)2D3 in magnesium deficient medium and those in standard magnesium medium were the same. In addition, dibutyryl cyclic adenosine monophosphate (dbc AMP) could restore expression of differentiation-related phenotypes inhibited by magnesium deprivation but not those inhibited by cyclohexamide, even though magnesium deprivation inhibited protein synthesis as much as did cyclohexamide. This suggests that magnesium-requiring step in HL-60 cell differentiation is different from that inhibited by cyclohexamide. PMID- 3032993 TI - Coordinate expression of c-myc, c-myb, and histone H4 genes in reversibly differentiating HL 60 cells. AB - The expression of oncogenes c-myc and c-myb in human leukemic cells HL 60 was compared to the expression of histone H4 gene, known to be cell-cycle dependent. Steady-state levels of mRNA transcribed from these genes were determined by simultaneous hybridization of Northern transfers with four probes, and the rates of gene expression were measured by nuclear transcription ("run-on") assays. Expression of genes c-myc, c-myb and histone H4 varied coordinately and in parallel with the rates of DNA synthesis, while the rates of total and ribosomal RNA synthesis, the expression of gene c-Ha-ras, unrelated to proliferation of these cells, and gene p 72, a constitutively expressed human gene, were unchanged. Further, the levels of c-myc and c-myb mRNA but not p 72 mRNA were higher in cell populations enriched for S phase cells. Thus, transcription of genes c-myc and c-myb in HL 60 cells appears to be linked to DNA replication in a manner previously demonstrated for core histone gene expression. PMID- 3032995 TI - Effects of diverse intracellular thiol delivery agents on glutathione peroxidase activity, the ratio of reduced/oxidized glutathione, and ornithine decarboxylase induction in isolated mouse epidermal cells treated with 12-O tetradecanoylphorbol-13-acetate. AB - Since the enhancement of the activity of the natural glutathione (GSH)-dependent antioxidant protective system of the epidermal cells appears to inhibit the oxidative challenge presumably linked to skin tumor promotion by 12-O tetradecanoylphorbol-13-acetate (TPA), we have compared the effectiveness of diverse intracellular thiol delivery agents as inhibitors of the effects of TPA on GSH metabolism and ornithine decarboxylase (ODC; L-ornithine carboxylase, EC 4.1.1.17) induction in isolated mouse epidermal cells. Here we report at a 2-mM concentration, the monoethyl and monomethyl esters of GSH, N-acetyl-L-cysteine, and L-2-oxothiazolidine-4-carboxylate are all significantly more effective than GSH in inhibiting the sharp decline in the intracellular ratio of reduced GSH/oxidized glutathione (GSSG), the prolonged decrease in GSH peroxidase (GSH:H2O2 oxidoreductase, EC 1.11.1.9) activity, and the induction of ODC activity caused by 1 microM TPA. Moreover, diethyldithiocarbamate prevents totally the initial drop in the GSH/GSSG ratio of TPA-treated cells and is the most potent inhibitor of TPA-decreased GSH peroxidase activity in relation with its remarkable 98% inhibition of TPA-induced ODC activity, suggesting that the potential antitumor-promoting activity of this compound in mouse skin may be far superior to that previously demonstrated by GSH in the initiation-promotion protocol. PMID- 3032997 TI - Transformation by the v-fms oncogene product: an analog of the CSF-1 receptor. AB - The product of the c-fms proto-oncogene is related to, and possibly identical with, the receptor for the macrophage colony-stimulating factor, M-CSF (CSF-1). Unlike the product of the v-erbB oncogene, which is a truncated version of the EGF receptor, the glycoprotein encoded by the v-fms oncogene retains an intact extracellular ligand-binding domain so that cells transformed by v-fms express CSF-1 receptors at their surface. Although fibroblasts susceptible to transformation by v-fms generally produce CSF-1, v-fms-mediated transformation does not depend on an exogenous source of the growth factor, and neutralizing antibodies to CSF-1 do not affect the transformed phenotype. An alteration of the v-fms gene product at its extreme carboxyl-terminus represents the major structural difference between it and the c-fms-coded glycoprotein and may affect the tyrosine kinase activity of the v-fms-coded receptor. Consistent with this interpretation, tyrosine phosphorylation of the v-fms products in membranes was observed in the absence of CSF-1 and was not enhanced by addition of the murine growth factor. Cells transformed by v-fms have a constitutively elevated specific activity of a guanine nucleotide-dependent, phosphatidylinositol-4,5-diphosphate specific phospholipase C. We speculate that the tyrosine kinase activity of the v fms/c-fms gene products may be coupled to this phospholipase C, possibly through a G regulatory protein, thereby increasing phosphatidylinositol turnover and generating the intracellular second messengers diacylglycerol and inositol triphosphate. PMID- 3032996 TI - Fusion-mediated microinjection of active amine and diamine oxidases into cultured cells: effect on protein and DNA synthesis in chick embryo fibroblasts and in glioma cells. AB - Serum amine oxidase and/or porcine kidney diamine oxidase were trapped within reconstituted Sendai virus envelopes, and retained their activity. The trapped enzymes that were detected by radioimmunoblots were microinjected into cultured cells by fusion. When diamine oxidase was microinjected into cultured fibroblasts of chick or rat embryos, a temporary arrest in protein and DNA synthesis was observed. The inhibitory effect was more significant when both serum amine oxidase and kidney diamine oxidase were microinjected into those cultured cells. Fibroblasts of either chick or rat embryos transformed by Rous sarcoma virus were more susceptible to the injected enzymes than the normal cultures, showing a complete arrest in protein and DNA synthesis within 4 hours. Similar results were obtained by microinjecting diamine oxidase into cultured glioma cells. The injected enzyme catalyzed the oxidation of intracellular polyamines. The resulting oxidation product (hydrogen peroxide and aminoaldehydes) apparently caused the arrest in the synthesis of macromolecules. PMID- 3032998 TI - High-performance liquid chromatographic method using a C18 column for the simultaneous separation of the products of decomposition and oligomerization of guanosine 5'-phospho-2-methylimidazolide. PMID- 3032999 TI - Investigation of perchlorate, phosphate and ion-pairing eluent modifiers for the separation of cephalosporin epimers. AB - The retention behavior of several pairs of 7 alpha- and 7 beta-cephalosporin epimers was investigated using perchlorate, phosphate and ion-pairing eluent modifiers. At pH 2.5, sodium perchlorate, sodium phosphate and sodium pentanesulfonate all provided separation of epimers with free 7-amino groups. When the 7-amino group was blocked, as in cephalexin and cefaclor, sodium perchlorate gave the best separation at pH 2.5. A tetrabutylammonium ion-pairing system at pH 7.0 provided separation of all epimer pairs containing a free carboxylic acid at the 3 position. Hydrophobic, residual silanol and ionic interactions were factors in the retention mechanism of the cephalosporins under the conditions investigated. An ionic interaction of perchlorate with the protonated amine of the cephalosporin was postulated as an explanation of the retention and selectivity effects observed with perchlorate as an eluent modifier. PMID- 3033000 TI - Reversed-phase high-performance liquid chromatography of tyrosine-containing metabolites of enkephalins using octanesulphonic acid as ion-pairing agent and radioactivity detection. PMID- 3033001 TI - Determination of acetylcholine and choline in perchlorate extracts of brain tissue using liquid chromatography-electrochemistry with an immobilized-enzyme reactor. PMID- 3033002 TI - Confirmation of cannabis abuse by the determination of 11-nor-delta 9 tetrahydrocannabinol-9-carboxylic acid in urine with high-performance liquid chromatography and electrochemical detection. PMID- 3033003 TI - Determination of catecholamines in urine (and plasma) by liquid chromatography after on-line sample pretreatment on small alumina or dihydroxyborylsilica columns. AB - The possibilities of the on-line pre-concentration of catecholamines on small columns packed with aluminium oxide (10 micron) or dihydroxyborylsilica (5 micron) prior to high-performance liquid chromatography (HPLC) were investigated. For both pre-concentration materials the influence of the pH of the mobile phase on the retention characteristics was studied for the catecholamines and some of their derivatives lacking the catechol and/or the amine function. The recovery of the catecholamines and the loading capacity on some pre-columns was then investigated. The compatibility of the pre-concentration system with several HPLC systems, viz., reversed-phase ion-pair, ion-exchange and reversed-phase ion-pair partition chromatography, was studied. Combined with amperometric or fluorimetric detection, the method was applied to the determination of free catecholamines in urine, using dihydroxybenzylamine or epinine as internal standards. An example of the determination of catecholamines in plasma containing 100 pg/ml of each catecholamine is also shown. PMID- 3033004 TI - Determination of kininase I and kininase II activities in human urine by high performance liquid chromatography. PMID- 3033005 TI - Oxytocin and vasopressin: distinct receptors in myometrium. AB - The binding characteristics of [3H]oxytocin [( 3H]OT) and [3H]lysine vasopressin [( 3H]LVP) to nonpregnant human myometrium were investigated. Binding of both radioligands was saturable, time dependent, and reversible. Whereas [3H]OT was found to bind to a single class of sites with high affinity [Kd, 1.5 +/- 0.4 (+/- SEM) nM] and low capacity [maximum binding (Bmax), 34 +/- 6 fmol/mg protein], [3H]LVP bound to two classes of sites, one with high affinity (Kd, 2.2 +/- 0.1 nM) and low capacity (Bmax, 198 +/- 7 fmol/mg protein) and another with low affinity (Kd, 655 +/- 209 nM) and high capacity (Bmax, 5794 +/- 1616 fmol/mg protein). The binding of the labeled peptides also displayed a marked difference in sensitivity to Mg2+ and guanine nucleotides. These differences in binding characteristics as well as the differences in potency of analogs in competing for [3H]OT and [3H]LVP binding indicate the presence of distinct receptors for OT and vasopressin in human myometrium. Pharmacological characterization of the high affinity binding sites for [3H]LVP indicated that these are of the V1 subtype. Although, as suggested by others, vasopressin and OT can bind to the same sites, the presence of distinct receptors for both peptides provides an explanation for the previously reported difference in myometrial responsiveness to OT and vasopressin. PMID- 3033006 TI - Corticotropin-releasing hormone stimulation test before and after transsphenoidal selective microadenomectomy in 30 patients with Cushing's disease. AB - Thirty patients with ACTH-dependent Cushing's disease were tested with CRH before and 7-10 days and 3-6 months after selective transsphenoidal adenomectomy. In 28 of 30 patients an adenoma was found, and in 22 (79%) clinical and endocrinological remission occurred. Preoperatively, the majority of the patients had basal and CRH-stimulated plasma ACTH levels that were markedly increased compared to those in normal subjects. On the basis of the CRH stimulation test and low dose (2 mg) dexamethasone suppression test results 7-10 days after surgery, these 30 patients were divided into 4 groups. Groups I, II, and III were patients in remission, as defined by undetectable, subnormal, or normal basal plasma ACTH and cortisol levels in addition to sufficient suppression of cortisol (less than 2 micrograms/dL) during the low dose (2 mg) dexamethasone suppression test. Patients in group IV were not in remission. In group I (n = 6), CRH failed to raise undetectable basal ACTH levels in the early postoperative period; however, 3-6 months later plasma ACTH did increase in response to CRH. In group II (n = 11), undetectable or low basal ACTH levels increased after CRH, and the increase was similar to that in normal individuals. In group III (n = 5), basal ACTH levels were normal, and the response to CRH was exaggerated, but all patients responded normally to the dexamethasone suppression test. The CRH induced ACTH increase in group III was significantly greater (P less than 0.003) than that in normal subjects, but was similar to that in patients not in remission in group IV (n = 6). Three to 6 months later, the ACTH response to CRH in group III was normal. In summary, the CRH test 7-10 days after surgery in patients with Cushing's disease indicated remission when there was no CRH-induced ACTH response or the response was normal (groups I and II). The test failed to predict remission in patients with an exaggerated CRH-induced ACTH response (groups III and IV). However, with regard to group II, the CRH-induced ACTH increase 1 week after selective adenomectomy indirectly supports the concept of CRH deficiency during hypercorticism and thus, in these patients as well as in group I, a pituitary origin of the disease. PMID- 3033007 TI - Functional uncoupling of the platelet alpha 2-adrenergic receptor-adenylate cyclase complex in the elderly. AB - Elderly humans demonstrate decreased responsiveness in several hormone-receptor systems, including adrenergic receptors. Studies of the beta-adrenergic receptor (beta-AR) system have shown that reduced beta-adrenergic sensitivity in the elderly may be due to reduced beta-AR affinity for agonists. To determine the mechanisms underlying altered alpha-adrenergic sensitivity in the elderly, we assessed the relationships between age and platelet membrane alpha 2-adrenergic receptor (alpha 2-AR)-binding properties, receptor-linked adenylate cyclase (AC) activity, and the affinity of the alpha 2-AR-AC complex for agonists in 18 young (mean age, 24 yr; range 19-34) and 13 elderly (mean age, 69 yr; range, 63-85) normal subjects. In platelet membrane preparations from elderly compared to young subjects, we found similar antagonist-binding properties and similar activity of the catalytic unit of platelet AC, as indicated by the cAMP response to sodium fluoride stimulation. However, mean epinephrine-mediated inhibition of sodium fluoride-stimulated platelet AC activity was less in the elderly [20 +/- 4% (+/- SEM) vs. 31 +/- 2% inhibition; P less than 0.005). In addition, platelet alpha 2 AR affinity for agonist was lower in the elderly, as indicated by the higher concentration of epinephrine needed to inhibit 50% of specific [3H]yohimbine binding (IC50, 3.2 +/- 0.6 vs. 1.4 +/- 0.3 microM; P less than 0.02). These data provide evidence that platelet membranes from elderly humans have decreased responsiveness to alpha-adrenergic stimulation, which can be attributed to reduced alpha 2-AR-AC affinity for agonists. Similarly to reported age-related alterations in beta-adrenergic receptor function, these results suggest that there is also functional uncoupling of the alpha 2-AR-AC complex in elderly humans. PMID- 3033008 TI - Sunscreens suppress cutaneous vitamin D3 synthesis. AB - Sunscreens block the cutaneous absorption of UV-B radiation and prevent sunburning, premature aging, and cancer of the skin. Inasmuch as UV-B radiation is also responsible for the photosynthesis of vitamin D3, we investigated the effect of sunscreens on the cutaneous formation of vitamin D3 in vivo and in vitro. Eight normal subjects, four of whom had been protected with the sunscreen para-aminobenzoic acid (sun protection factor 8), were exposed to one minimal erythema dose of UV radiation. The mean serum vitamin D3 concentration increased from 1.5 +/- 1.0 (+/- SEM) to 25.6 +/- 6.7 ng/mL in unprotected subjects, whereas it was 5.6 +/- 3.0 and 4.4 +/- 2.4 ng/mL at these times in the subjects who were protected with para-aminobenzoic acid. Para-aminobenzoic acid also prevented the photoisomerization of 7-dehydrocholesterol to previtamin D3 in human skin slices in vitro. These results indicate that the sunscreen interferred with the cutaneous production of vitamin D3. PMID- 3033009 TI - Effect of ovine corticotropin-releasing hormone administered during insulin induced hypoglycemia on plasma adrenocorticotropin and cortisol. AB - The factors that mediate the hypothalamic-pituitary response to hypoglycemia in man are unknown. To investigate the role of CRH in the plasma ACTH response to hypoglycemia, two different doses of ovine CRH (oCRH) were given to normal men during insulin-induced hypoglycemia. We hypothesized that if the endogenous CRH response to hypoglycemia were less than maximally stimulating, administration of oCRH during hypoglycemia would result in a greater peak plasma immunoreactive (IR) ACTH response. Six normal men were given 1) 0.15 U/kg regular insulin, iv; 2) insulin plus 1 microgram/kg oCRH, iv, 5 min after serum glucose fell to 40 mg/dL or less; and 3) oCRH alone. The degree and duration of hypoglycemia were the same when insulin was given alone or with oCRH. Plasma IR-ACTH after insulin alone and insulin plus oCRH rose at the same rate to similar peaks of 226 +/- 37 (mean +/- SEM) and 213 +/- 53 pg/mL, respectively, both of which were greater (P less than 0.05) than the peak plasma IR-ACTH after oCRH alone (61 +/- 19 pg/mL). The peak plasma IR-cortisol levels after insulin alone (24 +/- 4 micrograms/dL), insulin plus oCRH (27 +/- 3 micrograms/dL), and oCRH alone (18 +/- 2 micrograms/dL) were not significantly different. In a second study, six normal men were given 0.15 U/kg regular insulin, iv; insulin plus 10 micrograms/kg oCRH, iv; and 10 micrograms/kg oCRH alone. Administration of oCRH 5 min after serum glucose fell to 40 mg/dL or less did not affect the degree or duration of hypoglycemia. Plasma IR-ACTH after insulin alone and insulin plus oCRH rose at the same rate to similar peaks of 258 +/- 14 and 290 +/- 33 pg/mL, respectively, both of which were greater (P less than 0.01) than the peak (54 +/- 6 pg/mL) after oCRH alone. After insulin alone, plasma IR-ACTH declined to baseline by 3 h. However, after insulin plus oCRH, plasma IR-ACTH fell gradually until 2 h, rose to a second peak at 2.5-3 h, and remained greater (P less than 0.01) than after insulin or oCRH alone for the 4-h duration of the study. The mean peak plasma IR-cortisol level after insulin plus oCRH (33 +/- 4 micrograms/dL) was similar to that after insulin alone (28 +/- 3 micrograms/dL), but was greater (P less than 0.05) than that after oCRH alone (18 +/- 2 micrograms/dL).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3033010 TI - Effects of continuous subcutaneous insulin infusion and intensified conventional therapy on peripheral and autonomic nerve dysfunction. AB - The objective of this study was to determine whether a favorable effect of short term continuous sc insulin infusion (CSII) therapy on the peripheral and autonomic nervous system could be maintained by subsequent intensified conventional treatment (ICT). Nine type I diabetic patients, aged 18-32 yr, who had been diabetic for 4-23 yr and had reduced nerve conduction velocities received CSII for 4 weeks and subsequently ICT for up to 26 weeks. Motor and sensory nerve conduction velocities (MNCV and SNCV) and heart rate variations during deep breathing (E/I ratio), during lying and standing (30/15 ratio), and during the Valsalva maneuver (Valsalva ratio) were measured before CSII and at intervals of 1, 2, 4, 6, 10, and 26 weeks. During CSII, MNCV and SNCV increased significantly (P less than 0.01), the E/I ratio improved in seven patients (P less than 0.05), the Valsalva ratio increased in eight patients (P less than 0.01), and the 30/15 ratio increased in five patients. The E/I ratio increased significantly earlier than the Valsalva ratio (P less than 0.025). During ICT, nerve conduction velocity slightly, though not significantly, decreased, and the results of the cardiovascular reflex tests also gradually declined. The hemoglobin A1 concentration before initiation of CSII and the diminution of the hemoglobin A1 concentration during CSII therapy were inversely correlated to the increase in MNCV (P less than 0.01 and P less than 0.05, respectively). In conclusion, CSII improved peripheral and autonomic nervous system function, but the improvement diminished somewhat during ICT. PMID- 3033012 TI - Lymphocyte responses to varicella zoster virus in the elderly. AB - Elderly subjects (age 75-95 years) immune to varicella zoster virus (VZV) were identified by the presence of serum IgG antibody. The frequency of lymphocytes in their blood which proliferated in varicella zoster virus antigen-stimulated cultures was 1:78,000 +/- 6600. This is less than the 1:14,000 +/- 2000 frequency of VZV-responsive lymphocytes in blood from younger adult (20-43 years) donors. Elderly donors' blood mononuclear cells were less efficient than those of younger adults at lysing VZV-infected fibroblasts but not K562 target cells. The lysis of VZV target cells by elderly donors' MNC increased to control levels in the presence of 10 U/ml of interleukin-2 (IL-2). These results suggest that mononuclear cells capable of killing VZV-infected target cells persist with aging but that reduced numbers of antigen-responsive and lymphokine-releasing T cells may limit their function. PMID- 3033011 TI - The significance of antilymphocyte antibodies in patients with acquired immune deficiency syndrome (AIDS) and their sexual partners. AB - Antilymphocyte antibodies have been demonstrated in autoimmune diseases, acute viral infections, and acquired immune deficiency syndrome (AIDS) by using either the conventional microlymphocytotoxicity or the double fluorescence technique. In the present study, we used both methods to detect the antilymphocyte antibodies and to characterize further their immunologic significance in patients with AIDS and their sexual partners. The results using the conventional microlymphocytotoxicity method demonstrated that 8 of 10 patients with AIDS and 6 of 10 partners had significant levels of antilymphocyte antibodies which were reactive with B and T cells at cold and warm temperatures. A significant loss in antibody activity following absorption with B, T, and Daudi cells and Staphylococcus aureus, but not platelets or red cells, indicated that these antibodies are not directed to HLA class I antigens but, rather, to antigens that are common to both groups of lymphocytes. There is a close association between antilymphocyte antibodies and lymphopenia in patients but not in partners. Antibodies against lymphocyte subclasses [helper (T4) and suppressor (T8)] were detected by the double fluorescence staining technique, which employs C6 deficient serum as a nonlytic source of complement, and demonstrated the binding of antibodies to target cells, in contrast to lysing of the target cells as in the microlymphocytotoxicity method. The results of this assay showed that antibodies were directed to both populations, and there was no correlation or association between the absolute numbers of peripheral T4 and T8 cells and the percentage of antibody binding. Taken together, there appear to be at least two kinds of antilymphocyte antibodies: lymphocytotoxic antibodies detected by the conventional microlymphocytotoxicity assay and noncytotoxic antibodies detected by the double fluorescence staining technique. The former may be responsible in part for the lymphopenia. The latter may alter lymphocyte function. The patients and partners who had antilymphocyte antibodies also had anti-HTLV-III antibodies, although there was not any close correlation between titers. These findings support the possibility that both types of antibodies occur as part of a generalized immune response, possibly stimulated by the same viral agent. PMID- 3033013 TI - Simple and specific enzyme immunoassay using monoclonal antibodies for serotyping human rotaviruses. AB - An enzyme immunoassay for serotyping human rotaviruses in stools and in cell culture was developed. Hyperimmune rabbit antisera to rotaviruses were used as capture antibodies, and rotavirus-neutralizing mouse monoclonal antibodies specific for serotypes 1, 2, 3, and 4 were used as detection reagents. Partial purification of monoclonal antibodies and inclusion of skim milk powder in antibody diluents contributed to assay specificity. The sensitivity of this assay was greater than that of a direct enzyme immunoassay in which rotaviruses of the appropriate serotype were adsorbed directly to the solid phase. When fecal extracts were concentrated threefold, this serotyping enzyme immunoassay was of equal specificity and approached the sensitivity of electron microscopy for rotavirus detection. This assay is simple and rapid and is suitable for serotyping the large numbers of isolates obtained from epidemiological studies and vaccine trials. PMID- 3033014 TI - Molecular probe for identification of medically important Candida species and Torulopsis glabrata. AB - A cloned DNA fragment from Candida albicans containing the gene for the protein actin was used to probe the molecular structure of the actin gene of several medically important yeasts (C. albicans, Candida stellatoidea, Candida tropicalis, Candida pseudotropicalis, Candida krusei, Candida parapsilosis, Candida guilliermondii, and Torulopsis glabrata). Whole-cell DNA from each species was digested with restriction endonucleases, electrophoresed on agarose gels, and transferred to nitrocellulose. Radioactively labeled C. albicans actin gene was hybridized to the DNA fragments on the nitrocellulose. The C. albicans probe produced a strong signal with all of the Candida DNAs tested, indicating considerable conservation of this gene. In addition, the actin genes of all of the species tested were found to have no internal EcoRI or SalI restriction sites. With the exception of C. guilliermondii, all of the species tested had a single internal HindIII recognition site. However, the location of flanking restriction sites was found to be species specific. For all of the enzymes tested, the locations of the flanking restriction sites in C. albicans and C. stellatoidea were identical; all of the other strains yielded fragments clearly distinct from one another. These differences provide a molecular tool for the differentiation of medically important Candida species. PMID- 3033015 TI - Detection of antibody to murine cytomegalovirus by enzyme-linked immunosorbent and indirect immunofluorescence assays. AB - We have compared murine cytomegalovirus (MCMV) antibody determination by enzyme linked immunosorbent assay (ELISA) and indirect immunofluorescence assay. A comparison of antibody detection with 146 serum samples at a 1:20 dilution showed 100% agreement (60 negatives and 86 positives) between the assays. There was close agreement of endpoint determinations of sera by both methods. After experimental MCMV infection, antibody to MCMV was detected by both assays as early as day 7, and high titers persisted as late as 6 months. In contrast to immunocompetent littermates, athymic nude mice did not develop antibody after infection. Mice lacking antibody detectable by ELISA were susceptible to lethal MCMV challenge. In a survey of animals from five commercial sources, MCMV antibody was not detected unless mice were experimentally infected. MCMV antibody determination by ELISA is a convenient method, comparable to the indirect immunofluorescence assay in sensitivity and specificity. PMID- 3033016 TI - Application of DNA typing methods to epidemiology and taxonomy of Candida species. AB - Methods are described for extraction of DNA from the yeast form of Candida spp., followed by digestion and electrophoresis of DNA fragments. The resulting gel patterns (greater than 100 bands) were used to type Candida isolates. Four intense bands identified, three of which are present in each isolate (6 to 7, 3.7 or 4.2, and 2.5 to 3 kilobases), appear to be DNA encoding the rRNA. The methods proved to be both simple and reproducible. The patterns were shown to be stable through several hundred doublings from multiple single colonies. A survey of isolates showed that, on the basis of similarity of gel patterns, several Candida species could be sorted into mutually exclusive groups, and subgroups could be created. Analyses of this survey suggested the possible epidemiologic and taxonomic applications of these methods. DNA typing methods appear to offer important potential advantages over phenotyping methods. The methods provide a base for further epidemiologic studies and for further development of techniques, such as the use of cloned probes for studies of DNA homology. PMID- 3033017 TI - Application of whole-cell DNA restriction endonuclease profiles to the epidemiology of Clostridium difficile-induced diarrhea. AB - Two patients in one hospital room acquired pseudomembranous colitis, one shortly after the other. The DNA restriction patterns of isolates from the patients and of four isolates from the environment were indistinguishable from one another and differed from isolates of other patients. Restriction endonuclease digest analysis appears to be a useful method for studying the epidemiology of Clostridium difficile. PMID- 3033018 TI - Suppression of the immune response by benzodiazepine receptor inverse agonists. AB - 24 h after administration of a single dose of the benzodiazepine receptor inverse agonists N'-methyl-beta-carboline-3-carboxamide (FG 7142) and 3-carbomethoxy-4 ethyl-6,7-dimethoxy-beta-carboline (DMCM), a profound suppression of the immune response was observed in rodents. This immunosuppression was manifest as a decrease in phytohemagglutinin (PHA) and concanavalin-A (Con-A) stimulated T cell proliferation in rats and mice administered FG 7142 and a decrease in allogeneic cytotoxic T lymphocyte activity in mice administered either FG 7142 or DMCM. The effects of FG 7142 were antagonized by the prior administration of Ro 15-1788, a benzodiazepine receptor antagonist. These findings demonstrate that the neural pathways subserved by benzodiazepine receptors can modulate immune function, and suggest that these receptors may be involved in the stress-induced modulation of immune function. PMID- 3033020 TI - 99mTc-pertechnetate uptake in parotid acinar cells by the Na+/K+/Cl- co-transport system. AB - 99mTc-Pertechnetate (99mTcO4-) has widespread clinical use in the diagnosis and evaluation of dysfunctions in many different tissues. However, despite the broad clinical application of this radionuclide, very little is known about the mechanism by which 99mTcO4- enters a cell. We report evidence here that 99mTcO4- shares the Na+/K+/Cl- co-transport system localized to the basolateral membrane of rat parotid acinar cells. 99mTcO4- uptake by these cells was quite rapid (t1/2 approximately 30 s), was completely inhibited by the loop diuretics furosemide and bumetanide, and was markedly dependent on the presence of Na+, K+, and Cl- in the extracellular medium. Relative to uptake measured in the presence of physiological extracellular salt concentrations (Hanks' salts), 99mTcO4- uptake was inhibited 80% by sodium replacement and 50% by potassium replacement. When Cl was replaced with the physiologically inert anion gluconate a threefold stimulation in 99mTcO4- uptake resulted. These observations provide strong evidence that 99mTcO4- can substitute for Cl- as a substrate for the Na+/K+/Cl- co-transporter and indicate that 99mTcO4- uptake by salivary glands (e.g., as seen with salivary scintiscans), and possibly by a variety of other tissues, reflects the functional activity of this co-transport mechanism. PMID- 3033019 TI - Antigen receptor genes as molecular markers of lymphoid neoplasms. PMID- 3033021 TI - Insulin receptor kinase in human skeletal muscle from obese subjects with and without noninsulin dependent diabetes. AB - We have studied the structure and function of the insulin receptors in obese patients with and without noninsulin dependent diabetes mellitus (NIDDM) and in nonobese controls using partially purified receptors from muscle biopsies. Insulin binding was decreased in obesity due to reduced number of binding sites but no differences were observed in insulin binding between obese subjects with or without NIDDM. The structural characteristics of the receptors, as determined by affinity labeling methods and electrophoretic mobility of the beta-subunit, were not altered in obese or NIDDM compared to normal weight subjects. Furthermore, the ability of insulin to stimulate the autophosphorylation of the beta-subunit and the phosphoamino acid composition of the phosphorylated receptor were the same in all groups. However, insulin receptor kinase activity was decreased in obesity using Glu4:Tyr1 as exogenous phosphoacceptor without any appreciable additional defect when obesity was associated with NIDDM. Thus, our data are supportive of the hypothesis that in muscle of obese humans, insulin resistance is partially due to decreased insulin receptors and insulin receptor kinase activity. In NIDDM the defect(s) in muscle is probably distal to the insulin receptor kinase. PMID- 3033022 TI - Mechanisms underlying the differential effects of ethanol on the bioavailability of riboflavin and flavin adenine dinucleotide. AB - Chronic alcoholism is associated with a high prevalence of riboflavin deficiency. Experiments were designed in an animal model to determine whether ethanol alters selectively the absorption of riboflavin and flavin adenine dinucleotide (FAD), the predominant dietary form of the vitamin. Rats received by gavage a liver homogenate to which either [14C]riboflavin or [14C]FAD was added with either ethanol or isocaloric sucrose solutions. Ethanol markedly diminished the bioavailability of [14C]FAD to a greater degree than that of [14C]riboflavin. Corroboration of an ethanol-impaired intraluminal hydrolysis of FAD was provided by using everted jejunal segments and measuring mucosal uptake of [14C]riboflavin together with nonradiolabeled FAD. In subsequent studies with mucosal cell extracts, ethanol markedly inhibited activities of FAD pyrophosphatase and flavin mononucleotide (FMN) phosphatase. These findings suggest that dietary sources of riboflavin (FMN and FAD) are not absorbed as well in the presence of ethanol than are vitamin preparations containing riboflavin, which is utilized more readily. PMID- 3033023 TI - New mechanism for glomerular injury. Myeloperoxidase-hydrogen peroxide-halide system. AB - Reactive oxygen species, particularly hydrogen peroxide (H2O2), participate in neutrophil-mediated glomerulonephritis. However, the mechanism of H2O2 neptrotoxicity is unknown. Myeloperoxidase (MPO), a neutrophil cationic enzyme that localizes in glomeruli, can react with H2O2 and halides to form highly reactive products. We tested the hypothesis that the MPO-H2O2-halide system may induce glomerular injury by infusing MPO followed by H2O2 in a chloride containing solution into the renal artery of rats. Controls received MPO or H2O2 alone. MPO-H2O2-perfused rats developed significant proteinuria, endothelial cell swelling, and epithelial cell foot process effacement, whereas control kidneys were normal. In the presence of free 125I, MPO-H2O2-perfused rats incorporated large amounts of 125I, localized to the glomerular basement membrane and mesangium by autoradiography, into glomeruli. Glomerular iodination was greatly decreased or absent in controls. The MPO-H2O2-halide system causes glomerular injury and may be important in neutrophil-mediated glomerulonephritis. PMID- 3033024 TI - Gene deletions correlate with the development of alloantibodies in von Willebrand disease. AB - Among all patients with von Willebrand disease (vWD), alloantibodies to von Willebrand factor (vWF) have been described only in severe vWD (type III). The relationship between the development of alloantibodies and the nature of the genetic lesion in vWD is not known. In hemophilia B, large deletions within the factor IX gene appear to correlate with the occurrence of alloantibodies, whereas in hemophilia A no such correlation is apparent. We have studied 19 patients with severe recessive vWD (type III) and 19 with autosomal dominant vWD (type I) by Southern blotting with probes encompassing the full 9 kilobases (kb) of the vWF cDNA. Two apparently unrelated patients were shown to have large deletions within the vWF gene. Both patients had severe vWD (type III) and were the only patients among those studied that had inhibitory alloantibodies to vWF. The extent of deletion was similar in both patients, corresponding to at least the 3'-7.4 kb of the vWF cDNA. The deletion in each patient was estimated to exceed 110 kb. In addition, the localization of the vWF gene to chromosome 12 was confirmed, and a homologous sequence on chromosome 22 was identified. PMID- 3033025 TI - Role of sorbitol accumulation and myo-inositol depletion in paranodal swelling of large myelinated nerve fibers in the insulin-deficient spontaneously diabetic bio breeding rat. Reversal by insulin replacement, an aldose reductase inhibitor, and myo-inositol. AB - Axo-glial dysjunction refers to the disruption of important junctional complexes that anchor terminal loops of myelin to the paranodal axolemma in diabetic human and animal peripheral nerve. Neither axo-glial dysjunction nor the preceeding acute localized paranodal swelling has been specifically attributed to discrete metabolic consequences of insulin deficiency or hyperglycemia. Two metabolic sequelae of hyperglycemia in diabetic nerve, sorbitol accumulation via aldose reductase, and (Na,K)-ATPase deficiency related to myo-inositol depletion, were explored as possible underlying causes of acute paranodal swelling in the spontaneously diabetic bio-breeding rat. 3 wk of insulin replacement, or therapy with an aldose reductase inhibitor or myo-inositol completely reversed paranodal swelling in sural nerve fibers after 3 wk of untreated insulin deficiency. These observations suggest that insulin deficiency and hyperglycemia cause reversible paranodal swelling, and ultimately poorly reversible axo-glial dysjunction, via the myo-inositol-related (Na,K)-ATPase defect rather than by the osmotic effects of sorbitol accumulation within nerve fibers. PMID- 3033026 TI - Variation of erythroid and myeloid precursors in the marrow and peripheral blood of volunteer subjects infected with human parvovirus (B19). AB - Infection of normal individuals with human parvovirus (B19) results in a mild disease (erythema infectiosum) but gives rise to aplastic crises in patients with chronic hemolytic anemias. The effects of this disease on hemopoiesis were investigated following intranasal inoculation of the virus into three volunteers. A typical disease ensued with a viremia peaking at 9 d. Marrow morphology 6 d after inoculation appeared normal but at 10 d there was a severe loss of erythroid precursors followed by a 1-2-g drop in hemoglobin, and an increase in serum immunoreactive erythropoietin. Erythroid burst-forming units (BFU-E) from the peripheral blood were considerably reduced, starting at the time of viremia and persisting for 4-8 d depending on the individual. Granulocyte-macrophage colony-forming units (CFU-GM) were also affected but the loss started 2 d later. Both CFU-GM and BFU-E showed a sharp overshoot at recovery. In the marrow, BFU-E and CFU-E were reduced at 6 and 10 d in the individual having the longest period of peripheral progenitor loss. In contrast, there was an increase in BFU-E and CFU-E in the subject with least change in peripheral progenitors. In the third subject, with an intermediate picture, there was a loss at 6 d but an increase at 10 d of erythroid progenitors. It is suggested that the architecture of the marrow might partially isolate progenitors from high titers of virus in the serum and individual variation in this respect might give the results observed. PMID- 3033028 TI - Distribution and size of thalamic neurons projecting to layer I of the auditory cortical fields of the cat compared to those projecting to layer IV. AB - The distribution of thalamocortical neurons projecting to layer I of the cat auditory cortical fields was examined by the horseradish peroxidase (HRP) method. After HRP injection into layer I of the primary auditory cortex (AI), HRP-labeled neuronal cell bodies were distributed mainly in the medial, dorsal, and ventrolateral divisions of the medial geniculate nucleus (MGN) and suprageniculate nucleus (Sg), and additionally in the lateral and medial divisions of the posterior group of the thalamus (Pol and Pom), lateroposterior thalamic nucleus (Lp), and nucleus of the brachium of the inferior colliculus (BIN). After HRP injection into layer I of the second auditory cortex (AII), labeled neurons were seen mainly in the medial, dorsal, and ventrolateral divisions of the MGN and Sg and additionally in the Pom, Lp, and BIN. After HRP injection into layer I of the anterior auditory field (AAF), labeled neurons were located mainly in the medial and dorsal divisions of the MGN, Sg, Pol, and BIN, and additionally in the ventrolateral divisions of the MGN, Pom, and Lp. After HRP injection into layer I of the dorsal part of the posterior ectosylvian gyrus (Epd), labeled neurons were observed chiefly in the medial and dorsal divisions of the MGN, Sg, and Lp and additionally in the ventrolateral division of the MGN, Pom, and BIN. After HRP injection into layer I of the ventral part of the posterior ectosylvian gyrus (Epv), labeled neurons were distributed chiefly in the medial and dorsal divisions of the MGN and Pol and additionally in the ventrolateral division of the MGN, Sg, and BIN. Thus no labeled neurons were found in the ventral division of the MGN after HRP injection into layer I of all auditory cortical fields examined in the present study. The average soma diameters of neurons that were labeled after HRP injection into layer I were statistically smaller than those of neurons that were labeled after HRP injection into layer IV. PMID- 3033027 TI - Novobiocin enhances alkylating agent cytotoxicity and DNA interstrand crosslinks in a murine model. AB - DNA-DNA crosslinks are the lethal cellular mechanism of bifunctional alkylating agent cytotoxicity. Novobiocin, an inhibitor of DNA topoisomerase II, impairs eukaryotic DNA repair of alkylating agent adducts and may increase the number of adducts and their resultant cytotoxicity in malignant cells. The effect of novobiocin on clonogenic survival and DNA crosslinking due to cisplatin (cDDP) and carmustine (BCNU) was studied. Novobiocin caused synergistic cytotoxicity in Chinese hamster ovary cells exposed to cDDP or BCNU. Novobiocin and cDDP increased the formation of DNA-DNA interstrand crosslinks six-fold greater than cDDP alone. The effect was schedule dependent. Novobiocin and cDDP or BCNU markedly reduced in vivo growth of a murine fibrosarcoma without increased host toxicity. As a modulating agent of cytotoxicity due to DNA-DNA crosslinking, novobiocin may enhance the clinical effectiveness of the alkylating agents in human cancer and offer insight into new therapeutic strategies. PMID- 3033029 TI - Action and location of neuropeptide tyrosine (Y) on hippocampal neurons of the rat in slice preparations. AB - The action of bath applied NPY (1-1,000 nM) was investigated on hippocampal slices of the rat with extra- and intracellular recording. Neuropeptide Y (NPY) at 10-1,000 nM caused a concentration-dependent, long-lasting reduction of excitatory postsynaptic potentials (EPSPs) in the hippocampal subfield CA1 and the area dentata, and an even stronger reduction of population spikes. Paired pulse experiments with low intensity, stimulation-evoked PSPs showed a marked increase in facilitation in the presence of NPY, indicating a presynaptic action. Spontaneous burst firing of CA1 pyramidal cells in low calcium, high magnesium medium was reduced, indicating a partially postsynaptic inhibitory action of NPY on their dendrites. Intracellular recording from CA1 somata during NPY administration revealed a reduction of the amplitudes of excitatory-inhibitory postsynaptic potential (EPSP-IPSP) sequences in the absence of changes in membrane potential and conductance. Accommodation of firing during long depolarizing pulses and afterhyperpolarizations were unchanged. The innervation pattern of NPY immunoreactive fibers in the same regions was studied in slices adjacent to the ones used for electrophysiology by using antisera against NPY and light and electron microscopy. There is a dense innervation of CA1 by NPY immunoreactive axons and terminals, particularly in the stratum moleculare. NPY immunoreactive neurons are present in the stratum oriens and pyramidale. The NPY labeled axons of the stratum moleculare participate in numerous synaptic contacts with the smaller dendritic elements in this layer, many of which belong to pyramidal neurons. These observations provide evidence for a dendritic NPY immunoreactive innervation of CA1 neurons, which is in keeping with the electrophysiological effects of NPY on pyramidal neurons.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3033030 TI - Quantitative analysis of the choline acetyltransferase-immunoreactive axonal network in the cat primary visual cortex: I. Adult cats. AB - The laminar distribution of cholinergic axons was analyzed quantitatively in the visual cortex of adult cats by using immunocytochemistry with a monoclonal antibody against choline acetyltransferase (ChAT). ChAT(+) fibers and varicosities were counted at different locations within area 17 and the distribution patterns in various animals were compared. Choline acetyltransferase immunoreactivity was localized in fine, varicose fibers, which were present in all layers of the visual cortex. The density of labeled fibers was highest in layer I, which contained 14% of all fibers and 19.5% of all varicosities, and decreased toward deeper layers. The number of varicosities decreased more markedly toward deeper layers than the frequency of fibers. These distribution patterns were very consistent, showing only slight intra- and interindividual variability. PMID- 3033031 TI - Pharmacology and significance of nonsteroidal anti-inflammatory drugs in the treatment of skin diseases. AB - Intensive studies of the molecular pathways involved in common inflammatory skin disorders, coupled with detailed pharmacologic evaluation of the responses of skin to the end products of these pathways, have resulted in a much clearer understanding of the mode of action of nonsteroidal anti-inflammatory drugs. In particular the development of lipoxygenase inhibitors is prompting intense interest in their possible role as anti-inflammatory agents in psoriasis and other dermatoses. Because of the potency of these and other classes of new anti inflammatory drugs, careful monitoring of their pharmacokinetics in individual patients, especially those at risk for adverse reactions, will prove necessary, especially in the early stages of treatment. Meanwhile, currently available nonsteroidal anti-inflammatory drugs have a limited but significant place in the treatment of certain dermatoses. Current experience of the high incidence of adverse reactions to existing nonsteroidal anti-inflammatory drugs suggests that this will be no less a problem with new agents under development. The skin is frequently involved in adverse reactions to this class of drug, and past experience suggests that cutaneous reactions are among the earliest unwanted side effects reported in a new drug of this type. The dermatologist, therefore, has an important responsibility to observe, document, and report such "early warning signs" to the appropriate licensing authority and the manufacturer. PMID- 3033032 TI - Cutaneous ultrastructural changes and photosensitivity associated with amiodarone therapy. AB - Amiodarone, an antiarrhythmic agent, is known to cause photosensitivity and cutaneous hyperpigmentation. Five patients taking this drug for periods of 1 to 48 months were studied. Skin biopsy specimens taken from a sun-exposed site were assessed by light microscopy, electron microscopy, and direct immunofluorescence. Three patients allowed comparative studies to be done on a biopsy specimen from non-sun-exposed skin. Light microscopy findings, including special stains, were not diagnostic of amiodarone-associated cutaneous changes. Electron microscopy, however, displayed distinctive intracytoplasmic inclusions in many cell types, some of which have not been reported previously. These inclusions represent phospholipid membranes associated with amiodarone or its metabolites as the result of a drug-induced lipidosis. Previous reports had postulated the inclusions were lipofuscin. Sun exposure may accelerate the formation of these intracellular deposits because they are more prominent in sun-exposed skin. Four of the above five cases, plus two additional patients, had symptoms compatible with a photosensitivity. Porphyrin assays were normal. Of the six patients phototested, three showed acute reactions to ultraviolet A (UVA) and ultraviolet B (UVB) and significant delayed reactions to UVA and/or UVB. The patients who had normal phototesting were on the drug for shorter periods than those with positive tests. PMID- 3033033 TI - Lethal genes surviving by mosaicism: a possible explanation for sporadic birth defects involving the skin. AB - A genetic concept is advanced to explain the origin of several sporadic syndromes characterized by a mosaic distribution of skin defects. It is postulated that these disorders are due to the action of a lethal gene surviving by mosaicism. The presence of the mutation in the zygote will lead to death of the embryo at an early stage of development. Cells bearing the mutation can survive only in a mosaic state, in close proximity with normal cells. The mosaic may arise either from a gametic half chromatid mutation or from an early somatic mutation. This concept of origin is proposed to apply to the Schimmelpenning-Feuerstein-Mims syndrome, the McCune-Albright syndrome, the Klippel-Trenaunay syndrome, the Sturge-Weber syndrome, and neurocutaneous melanosis. Moreover, this etiologic hypothesis may apply to two other birth defects that have recently been delineated, the Proteus syndrome (partial gigantism of hands or feet, hemihypertrophy, macrocephaly, linear papillomatous epidermal nevus, subcutaneous hemangiomas and lipomas, accelerated growth, and visceral anomalies), and the Delleman-Oorthuys syndrome (orbital cyst, porencephaly, periorbital appendages, and focal aplasia of the skin. PMID- 3033034 TI - Genetic association between nest building and brown adipose tissue thermogenesis in female house mice. AB - Mice selectively bred for either high or low levels of thermoregulatory nest building were cold-acclimated (5 degrees C) for 3 weeks without nesting material; then body weight and food intake were measured. The mice selected for low nest building (Lows) of both sexes showed lower feed efficiencies than the high nest building mice (Highs), although their body weights were not significantly different (Table 1). This adds to a large body of evidence which suggests that nest building and feed efficiency were influenced by a common mechanism (Lacy et al. 1978; Sulzbach and Lynch 1984; Lynch et al. 1981; Lynch and Roberts 1984). Brown adipose tissue mitochondrial GDP binding and cytochrome c oxidase activity were measured in the above mice. In females, the Lows had 100% higher levels of total GDP binding than the Highs, while no difference between the lines was seen in males. Thus in the High females, lower energy expenditure through brown fat thermogenesis may account for their greater feed efficiency. In males, the genetic differences in feed efficiency must be due to differences in either thermogenesis in tissues other than brown fat, or mechanisms which reduce heat loss. PMID- 3033036 TI - Thermodynamic analysis of precisely measured oxygen equilibria of tench (Tinca tinca) hemoglobin and their dependence on ATP and protons. AB - Precise oxygen equilibria including extreme, high and low saturation values were determined for hemoglobin (Hb) from the freshwater teleost Tinca tinca at three temperatures, each at two pH levels and in the presence and absence of the erythrocytic cofactor ATP, at twofold molar excess over Hb. Analysis of the data in terms of Adair's successive oxygenation theory shows that in the absence of ATP, each of the four oxygenation steps are exothermic, but that net heat release decreases as pH falls from 8.2 to 7.4. ATP greatly depresses the temperature sensitivity of oxygenation particularly at physiological erythrocytic pH, where endothermic cofactor dissociation finds expression in a reverse temperature sensitivity for binding of the 3rd oxygen molecule to the tetrameric Hb. Enthalpy (delta Hi) and entropy (delta Si) changes of oxygenation vary with oxygenation step, i, as well as with pH and ATP addition, but the variations of delta Hi are similar to those of delta Si reflecting enthalpy-entropy compensation. The data show that the cooperative effects in tench Hb can be dominated either by entropic or enthalpic contributions, depending on the experimental condition and the oxygenation step. PMID- 3033037 TI - Effects of potassium carbonate and sodium bicarbonate on rumen function in lactating Holstein cows. AB - Twelve multiparous lactating Holstein cows were used to compare effects of 1) no buffer, 2) 1.5% sodium bicarbonate, 3) 1.25% potassium carbonate, or 4) 1.85% potassium carbonate in total diet on rumen environment and liquid turnover, dry matter intake and digestibility, milk yield and composition, and blood acid-base balance. Cows fed buffered diets had greater dry matter intake and greater digestibility of dry matter, acid detergent fiber, and neutral detergent fiber than controls. Rumen pH was higher in cows fed buffers than in controls 2 to 4 h postfeeding, but buffered diets were not different. Rumen volume, osmolality, and liquid turnover were unaffected by dietary treatment. Molar percentage of rumen acetate was greater, propionate was less, and acetate:propionate ratio was greater in cows fed 1.85% potassium carbonate compared with other treatments. There were no treatment effects on milk yield, although milk fat percentage tended to be greater in buffered diets. Blood acid-base balance was not altered. Cows fed diets containing potassium carbonate performed similarly to those fed sodium bicarbonate. No adverse effects of potassium carbonate on rumen function or environment were observed. Potassium carbonate is an acceptable buffer and serves as a potassium supplement. PMID- 3033035 TI - Changes in neck muscles in Swedish reindeer bucks during rutting season. AB - Three neck muscles in Swedish reindeer bucks have been studied before and during the rutting season. These were M. splenius, M. sternocephalicus and M. brachiocephalicus. For comparison, M. longissimus dorsi was chosen. Fibre composition and fibre size were studied in the four muscles as also was the metabolic potential of three enzymes, representing respiratory chain (cytochrome oxidase), beta-oxidation of fatty acids (3-hydroxyacyl-CoA dehydrogenase) and anaerobic glycolysis (lactate dehydrogenase). The extreme increase in size of certain muscles in the neck in connection with the rutting season (e.g. sternocephalicus, which increases from 250 g to 1,500 g) was to a great extent due to an increase in fibre size. In splenius, all three fibre types studied increased (I, IIA, IIB); in brachiocephalicus, mainly IIA and IIB; and in sternocephalicus, only the IIB. No corresponding fibre increase could be found in longissimus dorsi. In splenius and sternocephalicus from bucks older than 54 months, 60-70% of the fibres were of type I, and in brachiocephalicus, only about 40%. In all muscles but one, oxidative capacity (cytochrome oxidase) and beta oxidation of fatty acids (3-hydroxyacyl-CoA dehydrogenase) decreased significantly during the rutting season. This indicates that purposes other than the enhancement of energy production by fatty acid oxidation must account for the enlargement of the neck muscles. PMID- 3033038 TI - Effects of neutral detergent fiber and roughage source on dry matter intake and milk yield and composition of dairy cows. AB - Data were from 20 experiments that utilized early to midlactation Holstein cows fed complete mixed diets or fed at constant forage:concentrate ratios. Within-cow diet comparisons (1688 cow-periods) were analyzed by least squares analysis of variance; mathematical model included experiment, cow in experiment, period, body weight, and source of roughage. Objectives were to determine relationships between neutral detergent fiber content of diet and milk yield and dry matter intake. Roughages and number of cow-periods were: sugarcane bagasse/silage (507), cottonseed hulls (504), corn silage (268), ground corrugated boxes (170), alfalfa/peanut hay (132), and others (107). Dry matter intake and estimated net energy intake had linear effects on milk yield and explained 21.6 and 24.0% of its residual variation; milk yield had curvilinear (quadratic) effect and explained 22.4% of dry matter intake residual variation. Interaction between neutral detergent fiber and source of roughage on milk yield, 4% fat-corrected milk, and dry matter intake resulted in reductions of 5.6, 5.6, and 13% in residual variations. Results suggest neutral detergent (% of dry matter) has greater effect on dry matter intake than on milk yield and its use in formulating diets for dairy cows will be within roughage source. PMID- 3033039 TI - [Cyclic adenosine-3',5'-monophosphate and serotonin in the tissues of rats at various stages of a neurodystrophic process]. PMID- 3033041 TI - Strategy in immunocompromised patients with pulmonary infiltrates. PMID- 3033040 TI - Prevention of colonization and respiratory infections in long-term ventilated patients by local antimicrobial prophylaxis. AB - In a randomized clinical trial the prophylactic effects of locally administered antimicrobials on quantitative colonization and respiratory infections were studied in intubated patients with an expected period of mechanical ventilation of greater than 6 days. Nineteen patients received 50 mg of polymyxin B and 80 mg of gentamicin distributed among nose, oropharynx and stomach at 6-h intervals, as well as 300 mg of amphotericin B in the oropharynx. Twenty untreated patients served as controls. In the control group colonization by respiratory pathogens was more common (oropharynx 19 vs 6 patients (p less than 0.001); trachea 19 vs 11 (p less than 0.01)), and the number as well as the count of the colonizing species was usually higher. Fourteen patients of the control group developed respiratory infections, including nine cases of pneumonia, as compared to four patients with prophylaxis, including one case of pneumonia (p less than 0.01). Pneumonia-associated deaths were prevented with prophylaxis; however, the overall mortality remained unchanged. Respiratory infections in the prophylaxis group were associated with organisms resistant to the agents used, but the overall occurrence of resistance was not increased, as compared to the control group. We conclude that unrestrained upper airway colonization by respiratory pathogens and respiratory tract infection were causally related. Local antimicrobial prophylaxis proved to be a highly effective strategy for the prevention of potentially life-threatening pneumonias in critically ill patients, but in the present study the host setting appeared to be the major determinant of outcome. PMID- 3033042 TI - Platelet binding studies in panic disorder. AB - Binding parameters of the alpha 2-adrenoceptor (using [3H]yohimbine) and the 5-HT uptake site (using [3H]imipramine) were measured in intact platelets in a group of 15 patients with a DSM-III diagnosis of panic disorder. When compared with an age- and sex-matched control population no differences in either Bmax or Kd were found. This result is discussed in light of the reports of platelet binding abnormalities in depression. PMID- 3033043 TI - Enalaprilat, a new parenteral angiotensin-converting enzyme inhibitor: rapid changes in systemic and coronary hemodynamics and humoral profile in chronic heart failure. AB - Systemic and coronary hemodynamic, metabolic and humoral effects of a new intravenous angiotensin-converting enzyme inhibitor, enalaprilat, were evaluated in 14 patients with chronic heart failure. Onset of hemodynamic action occurred within 15 minutes and persisted for 6 hours. At the time of peak effect, there was a significant reduction in mean arterial pressure (-21%) and pulmonary capillary wedge pressure (-33%). Systemic vascular resistance decreased by 32% and stroke volume index increased by 20%. These systemic hemodynamic changes indicate improved left ventricular function. There was a substantial sustained reduction in rate-pressure product initially without a change in coronary sinus blood flow or myocardial oxygen consumption. There was also reduced myocardial oxygen extraction and augmented coronary sinus oxygen saturation at 30 minutes and 1 hour. In three patients, abnormal myocardial lactate extraction, present before enalaprilat, changed to uptake after enalaprilat, indicating amelioration of myocardial ischemia that was not clinically manifest. Systemic catecholamine levels and myocardial catecholamine balance did not change. Plasma renin activity increased and plasma aldosterone decreased. These findings suggest that enalaprilat produces inhibition of the angiotensin-converting enzyme and consequent beneficial systemic hemodynamic changes in heart failure. In some patients with heart failure, silent myocardial ischemia at rest can occur and can be alleviated with enalaprilat. Decreased myocardial oxygen extraction, increased coronary sinus oxygen saturation and lack of expected decrease in coronary sinus blood flow despite reduced rate-pressure product suggest transient coronary vasodilation by enalaprilat. PMID- 3033045 TI - The role of dietary carbohydrate in the decreased glucose tolerance of the elderly. AB - In this study we have attempted to evaluate the role of dietary carbohydrate (CHO) in the decreased glucose tolerance of aging. Eighteen healthy young (mean age, 27 +/- 1 standard error of the mean) (SEM) and 18 old (71 +/- 1 SEM) subjects matched for relative weight and socioeconomic group were studied. Oral glucose tolerance tests were performed while eating ad lib diet and after being given high (85%), medium (45%), and low (20% to 30%) CHO formula diets for at least three days. A three- to seven-day food record was used to determine the major nutrients including CHO and fiber intake in the ad lib diet at home. Our older subjects consumed significantly less total calories and slightly less carbohydrate but there was no difference in dietary fiber, protein, and fat intake. When studies were performed on matched CHO diets, decreased glucose tolerance was present in the older group. However, the mechanism(s) involved may vary with dietary CHO and with age. During the low CHO formula diet and the medium CHO ad lib diet, impaired insulin secretion was prominent in the elderly but was not present in the young. During the high CHO formula diet, insulin response was adequate in both old and young, but decreased glucose tolerance persisted in the older group, suggesting that insulin resistance may be the major contributing factor. We conclude that decreased glucose tolerance in the elderly is modified by CHO intake, but is present even when dietary CHO variability is acutely controlled.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3033044 TI - Regulation of coronary artery tone in relation to the activation of signal transductors that regulate calcium homeostasis. AB - Muscle tone of coronary arteries is regulated by free calcium concentration in the myoplasm. Various agonists, autacoids and putative peptides modify the calcium concentration directly or through actions of second messengers (signal transductors), such as cyclic guanosine monophosphate (GMP), cyclic adenosine monophosphate (AMP), inositol 1,4,5-trisphosphate, diacylglycerol or calmodulin. For example, acetylcholine (in the presence of intact endothelium cells), alpha human natriuretic peptide or nitrate compounds increase the amount of cyclic GMP and isoproterenol, prostacyclin (prostaglandin I2) or vasoactive intestinal polypeptide increases cyclic AMP. Both cyclic nucleotides reduce free calcium concentration. On the other hand, acetylcholine (in the presence or absence of endothelium cells), norepinephrine or thromboxane A2 increases inositol 1,4,5 trisphosphate and diacylglycerol, thus causing the increase in the free calcium concentration, whereas vasoactive intestinal peptide and alpha-human natriuretic peptide reduce them. Calmodulin acts as an internal calcium receptor for regulation of the contractile machinary. Regulation of calcium homeostasis in relation to the muscle tone in the coronary arteries including other vascular tissues is discussed together with the role of second messengers. PMID- 3033046 TI - Muscarinic receptors of rat parotid gland enlarged by gland ablation and bulk diet. Effects of denervation. AB - The density of muscarinic binding sites was increased 10% in the rat parotid gland enlarged (2 times control) as a result of ablation of the submandibular sublingual glands and maintenance of rats on bulk diet (50% inert cellulose plus 50% solid chow) for 4 weeks. When either the parasympathetic or sympathetic innervation to the gland was unilaterally removed at the time of submandibular sublingual ablation and introduction of the bulk diet, the density of muscarinic receptors showed an even greater increase from levels of innervated glands of chow-fed controls (29%); with removal of both nerves, the increase was 39%. A 36% increase in cyclic guanosine monophosphate levels accompanied the increase in receptors of the enlarged gland, but when the parotid was denervated, there was no change in cyclic GMP. Absence of either or both nerves led to a maximal decrease of 24-29% in density of muscarinic receptors of parotid gland of chow fed controls, but to no change in cyclic GMP levels. While autonomic influences mediate the changes in density of muscarinic receptors of parotid gland of chow fed rats, some additional factor is apparently involved in their increase in the enlarged gland. PMID- 3033047 TI - 1,10 phenanthroline, a metal chelator, protects against alloxan- but not streptozotocin-induced diabetes. AB - The iron chelator, 1,10-phenanthroline (Phen), which inhibits hydroxyl radical formation, was tested in vitro and in vivo against alloxan and streptozotocin (STZ) cytotoxicity. Phen injection reduced the severity of alloxan-induced diabetes in rats and attenuated alloxan-induced toxicity in human fibroblasts (VA 13 line) in culture. These protective effects were not observed against STZ toxic action. These results are consistent with the hypothesis that hydroxyl radicals, generated via an iron-catalyzed reaction, induce the alloxan but not the STZ diabetogenic effects. PMID- 3033049 TI - Lipid hydroperoxides initiate the autoxidation of sulfite with concomitant production of superoxide. AB - The inhalation of SO2 or the ingestion of beverages or food containing sulfite as a preservative has been associated with exacerbations of obstructive pulmonary disease. In this study it is demonstrated that 15-HPETE, a likely component of the lung's inflammatory response, can initiate the autoxidation of sulfite. Since 1 mol of 15-HPETE can initiate the consumption of 3 mol of oxygen and 6 mol of sulfite, it is likely that a free radical chain mechanism is operative. Direct evidence for the production of relatively small quantities of superoxide and indirect evidence for the production of lipid sulfonates are presented. It is possible that an intermediate free radical or product (as lipid sulfonate) is involved in the bronchospastic response to inhaled SO2. PMID- 3033048 TI - Photosensitization by antitumor agents 2: anthrapyrazole-photosensitized oxidation of ascorbic acid and 3,4-dihydroxyphenylalanine. AB - A novel anthrapyrazole anticancer agent has been examined for photosensitizing properties. Illumination of the anthrapyrazole and ascorbic acid with blue light in aerated aqueous solutions causes SOD and catalase-sensitive oxygen consumption, indicating involvement of both superoxide radical and hydrogen peroxide in this process. Electron paramagnetic resonance showed that the ascorbyl radical is also produced during the photooxidation. When 3,4 dihydroxyphenylalanine (Dopa) is used as a substrate, production of hydrogen peroxide is evidenced by catalase-sensitive oxygen consumption. Generation of hydroxyl radicals during illumination of the drug and ascorbic acid (or Dopa) in the presence of catalytic amounts of the Fe(III)/EDTA complex is demonstrated using EPR and spin-trapping techniques. PMID- 3033050 TI - Premarin priming does not alter growth hormone release following exercise. AB - We evaluated the usefulness of premarin priming on exercise induced growth hormone release and the value of combining several growth hormone screening agents in a large population of prepubertal children. Two hundred five short healthy prepubertal children growing below the 5th percentile in height were studied. One hundred forty-four were screened with exercise following glucose ingestion, while 61 were primed with estrogen prior to glucose and exercise testing. Premarin priming did not significantly increase the number of our patients who responded to exercise nor to glucose; 86% and 88.5% of non-primed and primed patients, respectively, responded with a growth hormone increase greater than or equal to 8 ng/ml following exercise and glucose. Glucose loading alone was not associated with a high enough growth hormone rise to rule out growth hormone deficiency in most of our children. Age (less than or equal to 5 yr) was associated with lower post-exercise growth hormone levels and a higher failure rate to testing in both primed and non primed children. Premarin priming does not seem to alter the growth hormone releasing capacity to exercise of prepubertal children. The combined use of exercise, glucose loading and premarin priming in a single screening test does not improve on the results obtained by growth hormone exercise screening alone. PMID- 3033051 TI - Thyroid and pituitary hormone responses to TRH in advanced nonalcoholic liver disease. AB - Basal T4, T3, TSH, prolactin and growth hormone levels were determined in several groups: patients with postnecrotic cirrhosis with hepatocellular carcinoma (n = 14); patients with postnecrotic cirrhosis but without hepatocellular carcinoma (n = 26); cholangiolar carcinoma (n = 9); and normal controls age-matched to within 5 yr of the liver disease subjects studied. In addition, TRH stimulation (400 micrograms TRH) was performed; TSH, prolactin and growth hormone responses over a 180-min time interval were evaluated for each subject. The responses observed varied between liver disease groups. The presence or absence of hepatocellular carcinoma was found to determine, at least in part, the type of response observed. Similarly, the presence or absence of hepatic encephalopathy determined, and/or reflected, at least in part, the type of response observed. Finally, for purposes of continuity, basal and TRH-stimulated levels of TSH, prolactin, growth hormone, T4 and T3 are compared in 3 settings of cirrhosis: alcoholic, nonalcoholic postnecrotic cirrhosis, and postnecrotic cirrhosis with hepatocellular carcinoma. PMID- 3033054 TI - Wheat and nonwheat dietary fibers. Is there a choice? PMID- 3033052 TI - Hirsutism caused by an androgen-producing ovarian tumor. A case of arrhenoblastoma. AB - A 34-year-old woman with excessive facial hirsutism and a 18-month history of amenorrhea was found to have an ovarian arrhenoblastoma of the intermediate type. The endocrine profile was determined before, during and after surgery. Determination of hormone levels indicated that, although both the delta 4 and delta 5 pathways were involved, the delta 4 pathway was probably predominant in androgen biosynthesis. Four weeks after removal of the tumor, the menstrual cycle was normalized and has subsequently remained regular. Six yr after operation, the patient had lost her hirsutism and neither radiological, palpatory nor endocrine signs of either recurrence or metastasis have been observed. PMID- 3033055 TI - Immune response in buffalo calves experimentally infected with buffalo-pox virus. PMID- 3033053 TI - Age and alpha-2 adrenergic regulation of plasma norepinephrine kinetics in humans. AB - To investigate whether the age-related elevation of plasma norepinephrine (NE) is due to impaired alpha-2 adrenergic inhibition of sympathetic nervous system (SNS) outflow, arterialized plasma NE kinetics were measured before and 120 to 140 min after 1.5 and 5.0 micrograms m/kg oral clonidine in 6 old (57 to 78 years) and 8 young (25 to 39 years) normotensive male volunteers. Baseline plasma NE levels were higher in old compared with young men (M +/- SEM, 355 +/- 58 vs. 197 +/- 22 pg/ml, p less than .02). Clonidine produced significant (p less than .05) dose related reductions in plasma NE, NE appearance rate, NE clearance, and mean arterial blood pressure (MAP) in both groups. There was no difference between old and young men in response to low dose clonidine. Following the higher dose, both groups had similar suppression of plasma NE (-51 +/- 7% vs. -58 +/- 2%, p greater than .05) and NE appearance (-60 +/- 6% vs. -62 +/- 2%, p greater than .05), but older men had a greater fall in NE clearance (-20 +/- 2% vs. -10 +/- 1%, p less than .003) and MAP (-28 +/- 3% vs. -10 +/- 4%, p less than .006). These findings suggest that sensitivity to alpha-2 receptor-mediated suppression of plasma NE and NE appearance is not diminished in elderly men. PMID- 3033056 TI - Entomological investigations of Japanese encephalitis outbreak in Gorakhpur and Deoria districts of Uttar Pradesh. PMID- 3033057 TI - Hepatic encephalopathy. Application of visual evoked responses to test hypotheses of its pathogenesis in rats. AB - A previous study of the patterns of visual evoked responses (VERs) in rats was interpreted as providing support for the synergistic neurotoxins hypothesis of the pathogenesis of hepatic encephalopathy (HE) due to fulminant hepatic failure (FHF). In contrast, other studies of the patterns of VERs in rabbits with different encephalopathies were interpreted as providing support for the concept that increased GABA-ergic tone may contribute to the neural inhibition of HE due to FHF. To attempt to resolve the discordant findings in these studies, additional studies of VERs have been undertaken in rats. To induce increased tissue levels of ammonia, mercaptans and fatty acids which are found in HE due to FHF, carefully predetermined doses of urease, dimethyldisulphide and octanoic acid were administered. The (pre-seizure) encephalopathy induced by these three agents was associated with abnormalities of the VER waveform that were fundamentally different from the abnormalities of the VER waveform associated with HE due to thioacetamide-induced FHF. However, the VER waveform in this model of HE due to FHF resembled closely that associated with pentobarbital-induced encephalopathy. These findings are in satisfactory agreement with those in the previous analogous studies in rabbits. They do not provide support for the synergistic neurotoxins hypothesis of the pathogenesis of HE, but are entirely consistent with increased GABA-ergic tone contributing to the neural inhibition of HE due to FHF. PMID- 3033058 TI - Cirrhosis and the aetiology of hepatocellular carcinoma. PMID- 3033059 TI - Clearance of hepatitis B surface antigen (HBsAg) after surgical resection of hepatocellular carcinoma. AB - The serum HBsAg in 4 chronic HBsAg carrier patients with hepatocellular carcinoma (HCC) cleared within 4-38 months after surgical resection of their hepatic tumors. Two patients developed anti-HBs. During the follow-up period from 21 to 28 months after HBsAg clearance, none of the patients regained positive serum HBsAg. Two patients who had had tissue HBsAg present, exclusively in the tumor, showed quick HBsAg clearance after resection. The other 2 patients had a delayed HBsAg clearance. One had tissue HBsAg in both the tumor and nontumoral liver. Only 1 patient had tissue HBsAg in the liver, but not in the tumor. During the same period of observation of 323 chronic HBsAg carriers, who had a variety of histologically-verified chronic liver diseases and were followed for more than 6 months, only 1 cleared the antigen. The spontaneous HBsAg clearance in our HBsAg carriers (1/323) was significantly lower than that (4/64) of HBsAg-positive HCC patients with tumor resection, P less than 0.004. The mechanisms of HBsAg clearance in HCC patients after surgical resection of tumors are discussed. PMID- 3033060 TI - In vitro and in vivo tumour localisation with a monoclonal antibody directed against a membrane antigen on the human hepatocellular carcinoma cell line PLC/PRF/5. AB - A monoclonal antibody, designated K-PLC1, has been produced to a tumour associated antigen on the cell membrane of the PLC/PRF/5 cell line. Using an indirect immunofluorescence technique this antibody produced membrane staining of three hepatocellular carcinoma (HCC) cell lines and it has positively stained 10 of 11 human HCC biopsy specimens. In vitro, 125I-labelled K-PLC1 binds specifically to PLC/PRF/5 cells, as shown by competitive inhibition experiments. Tumours derived from the PLC/PRF/5 cell line were grown in nude mice and groups of tumour-bearing animals were injected with either [125I]K-PLC1 or [125I]mouse IgG, and then killed at 1, 4 or 7 days post injection. Bound radioactivity was counted in a variety of solid organs. Tumour:liver ratios for K-PLC1 were greater than those for mouse IgG at each time point, the differences being greatest on day 4 (ratio K-PLC1 4.4 +/- 0.93, ratio mouse IgG 1.53 +/- 0.60, mean +/- SD, P less than 0.05). The amount of [125I]K-PLC1 bound was greater in the tumour than in any other solid organ, the differences again being maximal on day four. Blood pool radioactivity however remained high throughout the study period. We conclude that anti-HCC monoclonal antibodies may be of value as immunodiagnostic and immunotherapeutic agents in human HCC. PMID- 3033061 TI - Effects of alpha-adrenergic stimulation and beta-adrenergic blockade on azygos blood flow and splanchnic haemodynamics in patients with cirrhosis. AB - The effects of beta-blockade with propranolol and of alpha-adrenergic stimulation with methoxamine, a powerful alpha-agonist, on azygos blood flow and on systemic and hepatic haemodynamics were investigated in 26 cirrhotic patients with portal hypertension. Beta-adrenergic blockade with propranolol (n = 12), evidenced by a significant reduction of heart rate (-17 +/- 1%, P less than 0.001) and cardiac index (-17 +/- 2%, P less than 0.001), caused a mild but significant decrease of hepatic venous pressure gradient (-10 +/- 2%, P less than 0.05) and a marked fall of azygos venous blood flow (-31 +/- 5%, P less than 0.05). Alpha-adrenergic stimulation with methoxamine (n = 14), manifested by a significant increase of mean arterial pressure (19 +/- 2%, P less than 0.001), mimicked the effects of propranolol on hepatic venous pressure gradient (-10 +/- 4%, P less than 0.05) and cardiac index (-11 +/- 2%, P less than 0.001). However, azygos blood flow was not significantly reduced by methoxamine (0.7 +/- 0.1 vs 0.6 +/- 0.1 l/min). On the contrary, hepatic blood flow was significantly reduced by methoxamine (-19 +/ 4%, P less than 0.01) but not by propranolol (-7 +/- 7%, ns). Similarly, in 8 patients who received methoxamine after being beta-blocked by propranolol, azygos blood flow, that was markedly reduced by beta-blockade, did not experience a further reduction but increased slightly by alpha-adrenergic stimulation, while hepatic blood flow, that was not reduced by propranolol, decreased significantly during the subsequent methoxamine infusion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3033062 TI - Double-immunolabeling systems for phenotyping of immune cells harboring bovine viral diarrhea virus. AB - Optimal staining conditions were defined for simultaneous detection of bovine viral diarrhea virus (BVDV) and mononuclear leukocyte surface antigens in tissue sections and cytospins. Because of the extreme lability of the virus antigens and the variable stability of the epitopes on the cell differentiation antigens, cryopreservation had to be used. This method gives slightly sub-optimal preservation of morphology. However, the specificity and sensitivity of the immunolabeling ensured reliable identification of the double-labeled cells, i.e., the phenotypic identification of virus-infected cells within the immune system. PMID- 3033063 TI - Effect of dehydrating agents on DNA organization in herpes viruses. AB - With routine procedures of Epon- or GMA-embedding and a stain specific for DNA, the nucleoid of mature herpes simplex virus-type 1 (HSV-1) assumes the well-known form of a short, compact, hollow cylinder or torus. A new, more complex organization of DNA filaments in encapsidated HSV-1 was found in infected cells after aldehyde fixation, methanol dehydration, and Lowicryl embedment. We have determined that it is the use of methanol as dehydrating agent that permits visualization of this internal structure. The same new spatial organization of DNA can be seen in Epon and GMA sections when methanol dehydration is used. This organization is lost in a methanol-ethanol sequence of dehydration but can be restored in an ethanol-methanol sequence. Dimethylsulfoxide (DMSO) is the only other agent among several reviewed here which resembles methanol in its effect on HSV-1 DNA. Methanol had the same effect on five subfamilies of the herpes group (HSV-1, HSV-2, CCV, CMV, CTHV) but did not alter the nucleoid ultrastructure in simian virus 40 (SV40) and adenovirus type 5 (Ad 5). Therefore, it may sometimes, but not always, provide additional information about the organization of biological structures. PMID- 3033065 TI - Segmental cable modelling of electrotonic transfer properties of deep superior colliculus neurons in the cat. AB - A segmental cable model of tecto-reticulo-spinal neurons of cat superior colliculus was constructed, based on detailed anatomical measurements from three neurons. The calculated membrane resistance for which the model best fitted the measured input resistance was 2,300-3,000 omega cm2. Electrotonic length of dendrites fell under 0.59-1.20 and 0.52-1.05 length constants, while the mean electrotonic length for the three cells averaged 0.91, 0.79, 0.81 and 0.80, 0.70, 0.71 (for sealed-end and open-end cable termination, respectively). Dendrite-to soma conductance ratios averaged 16.0, 10.7, 7.5 and 21.3, 14.1, 11.4 for the two different end conditions, respectively. Synaptic efficacy was estimated by the transfer of steady-state voltage or current reaching the soma from distal dendritic locations. While voltage transfer was less than 1%, almost 60% of injected current (or charge) reached the soma. Analysis of voltage transients recorded experimentally in TRSNs demonstrated considerable difference between parameters derived from either equivalent-cylinder model or segmental cable model. The obvious deviations of TRSNs both in anatomical and electrotonic respect from the assumptions of the equivalent-cylinder model indicate that the detailed cable model will give a more appropriate description of these neurons. The significance of the estimated electrotonic parameters for the particular burst generation mechanism of TRSNs is discussed. PMID- 3033064 TI - Quantitative immunoelectron microscopic localization of (Na+,K+)ATPase on rat exocrine pancreatic cells. AB - Distribution of (Na+,K+)ATPase in rat exocrine pancreatic cells was investigated quantitatively by immunoelectron microscopy using the post-embedding protein A gold technique. We found that in acinar and duct cells (Na+,K+)ATPase exists on both the luminal and the basolateral surfaces, with higher particle density on the luminal surface (4.4 times in the acinar cells and 5.6 times in the duct cells). According to Bolender (J Cell Biol 61:269, 1974), the luminal surface represents only 5% of the total cell surface of an average pancreatic acinar cell. It is roughly estimated, therefore, that approximately 80% of the plasma membrane (Na+,K+)ATPase in the acinar cells exists on the basolateral surface. When the acinar and duct cells were compared, more than twice as many particles were found on acinar cells than on duct cells. The enzyme existed on all the cell surfaces, preferentially on the microvilli or on the cell membrane folds, and no clustering was detected. We suggest that the (Na+,K+)ATPase on the basolateral surface is mainly responsible for the extrusion of a large number of sodium ions that are incorporated into the cytoplasm accompanying the secondary active transport of various organic substances and inorganic ions, whereas that on the luminal surface is responsible for active extrusion of sodium ions that are partially responsible for the fluid secretion of the pancreatic cells. PMID- 3033066 TI - Altered structures in the cytoplasm of the ependymal cells next to the periventricular nucleus of the turtle Mauremys caspica. AB - In the apical cytoplasm of some ependymal cells of the Hypothalamic Periventricular Nucleus of the turtle Mauremys caspica large amount of cell structures of difficult diagnose are found. Their morphology is variable seeming to correspond to cell organelles in a process of degeneration, characteristic feature of the high metabolic activity in the ventricular barrier. Their possible physiological significance is discussed in the present work. PMID- 3033067 TI - Evaluation of a latex agglutination test for rapid detection of rotavirus in faecal specimens. AB - Two methods for detecting rotaviruses (latex agglutination, electron microscopy) have been compared on 80 faecal samples. These samples were obtained from infants between the age of four and 30 months hospitalized for acute gastroenteritis in the Children's Hospital, Karl Marx University at Leipzig, in 1982. Complete agreement among the two techniques was found in 75 specimens. Sensitivity of latex agglutination could be estimated at 95%, the specificity also at 95%. Only one sample reacted nonspecifically. Performance of the latex agglutination proved quite simple. The results indicate that latex agglutination is suitable for rapid screening of rotavirus induced gastroenteritis in clinical practice thus enabling the rate of nosocomial rotavirus infections in children's hospitals to be reduced. PMID- 3033068 TI - On the pathogenesis of spontaneous parvovirus infection. AB - The internal organs of 21 beagles spontaneously infected with parvovirus were examined histologically and in 10 of the dogs immunofluorescence examination was also performed. The study showed that the pathological process had started in the small intestine and from there the viral agent had spread through the regional lymph nodes into the other lymphatic and the haemopoietic organs causing there depletion predominantly of lymphocytes and arrest of haemopoiesis. Four dogs displayed conspicuous oedema of the media of the arterioli in the liver. In the cytoplasm of probably Kupffer's cells specific fluorescence was present in all the 10 dogs examined. On the other hand neither viral antigen nor histological changes were found in kidneys. Our observations suggest that spontaneous infection of dogs with parvovirus takes place per os. Parvovirus is not eliminated from the organism with urine. PMID- 3033069 TI - Activation of immature cortical thymocytes through the T11 sheep erythrocyte binding protein. AB - Two major pathways, the T cell receptor and the T11 alternate pathway, allow for T cell activation. In the human thymus, the T cell antigen receptor complex is reduced or absent on immature thymocytes, whereas the T11 glycoprotein is present at high cell surface density on all thymocytes. To determine whether activation through the T11 pathway induces similar or different changes in mature and immature thymocytes, we fractionated thymocytes according to their surface expression of the T3-T cell receptor (T3/Ti) complex. We report that two populations, one with high and one with low T3/Ti expression, can be activated through the T11 pathway to undergo nuclear activation and express IL 2 receptors. Moreover, in the absence of accessory cells, only the most mature population, expressing high T3 density, could be induced to proliferate, whereas the subset representing immature cortical thymocytes required accessory cells for proliferation. These findings suggest that the cellular microenvironment may have a critical role in regulating the activation of immature cortical thymocytes and that this cell population may not represent "nonfunctional" dead end cells, but rather a valid intermediate in human thymic differentiation. PMID- 3033071 TI - Retrovirus infection alters growth factor responses of T lymphocytes. AB - A murine helper/inducer T cell clone, D10.G4, has been infected with Kirsten murine sarcoma virus (KiSV) pseudotyped with an amphotropic murine leukemia virus. The resultant Ki-ras-expressing lines (KiSV-D10) remain dependent on exogenous factors for continued growth but display distinctly different mitotic responses to certain cytokines as compared to the uninfected parent clone. Unlike the parent D10.G4 cells, these KiSV-D10 cells can be maintained in vitro indefinitely in the presence of recombinant interleukin 2 (IL 2), and they all display a maximal proliferative response to purified or recombinant interleukin 1 (IL 1). The IL 1-induced proliferation is shown not to be dependent or secretion of the T cell autocrine growth factors IL 2 or B cell stimulatory factor-1 (BSF 1). The KiSV-D10 lines show certain differences from one another and parent D10.G4 cells in their secretory and proliferative responses to T cell receptor- and BSF-1 mediated signals. These viral oncogene-expressing T cell lines, which remain responsive to and dependent on physiologic growth factors, should prove valuable for analyzing the mechanisms of action of single oncogenes and the intracellular events in T lymphocyte activation. PMID- 3033070 TI - Interleukin 1 (IL 1) as a mediator of crystal arthritis. Stimulation of T cell and synovial fibroblast mitogenesis by urate crystal-induced IL 1. AB - We reported before that monosodium urate (MSU) crystals were potent stimulators of endogenous pyrogen (EP) production from human and rabbit mononuclear phagocytes, and proposed that this property of MSU crystals may be important in the pathogenesis of gout. EP activity is now attributed to interleukin 1 (IL 1) peptides but IL 1 is not the only pyrogenic monocyte-derived cytokine, since both interferon-alpha (alpha-IFN) and tumor necrosis factor (TNF) are also pyrogenic in rabbits. Using a T cell comitogenic assay based on a murine helper T cell clone that does not respond to IFN or TNF, we now report the release of IL 1 activity from human blood monocytes and synovial fluid mononuclear cells (MNC), following stimulation with MSU crystals. MSU-induced supernatants with IL 1 activity were neutralized with rabbit antiserum to human IL 1 and also stimulated the growth ([3H]thymidine incorporation) of long-term fibroblast-like cell lines derived from human synovial rheumatoid exudate. Two other crystals associated with articular inflammation were tested: hydroxyapatite was a much less potent stimulus compared with MSU crystals, and calcium pyrophosphate dihydrate did not stimulate IL 1 release from human monocytes or synovial fluid MNC. As a model for the inflammatory consequences of acute and chronic overproduction of IL 1, gout is the only sterile inflammatory disease where the local and systemic pathology is compatible with such overproduction; raised IL 1 levels have been found at the site of inflammation, and a necessary etiologic agent, crystalline urate, has been shown unequivocally to be a direct activator of mononuclear IL 1 release. PMID- 3033072 TI - Quantitative analysis of the T cell response to antigen and planar membranes containing purified Ia molecules. AB - Planar lipid membranes containing the purified Ia molecule E beta k:E alpha k can present a peptide antigen derived from cytochrome c to the T cell hybridoma 2B4.11. The incorporation of E beta k:E alpha k into planar membranes was linear over a 120-fold range of Ia molecule concentrations, permitting the dependence of the T cell response on the Ia molecule concentration to be examined. As the Ia molecule concentration was increased in the planar membranes, two parameters changed: less antigen was needed to stimulate the T cells, and the plateau response seen at functionally saturating concentrations of antigen increased. The antigen sensitivity was analyzed by plotting the antigen concentration (log2) required to stimulate the release of 10 U of IL 2 from the T cells as a function of the Ia molecule concentration (log2). If the T cell recognized a simple unit of one antigen molecule and one Ia molecule, this plot should have generated a straight line with a slope of -1. Surprisingly, a line with a slope of -2.04 X/divided by 1.12 was observed, suggesting that the T cell might recognize one antigen molecule and two Ia molecules. This complexity, however, resulted from changes in the maximal response achieved at different Ia molecule concentrations. A similar phenomenon was observed when the Ia molecule concentration was decreased in cultures containing splenic antigen-presenting cells (APC) by the addition of an anti-E beta k:E alpha k monoclonal antibody, or the use of [B10.A(4R) X B10.PL]F1APC. The Ia molecule concentration can therefore be limiting for T cell hybridomas in cultures containing normal APC and functionally saturating amounts of antigen. When the planar membrane data were normalized to the maximal response to eliminate the effect of the changing plateau response, the resulting plot generated a line with a slope of -1.17 X/divided by 1.11. These results suggest that the sole stimulatory signal for this T cell hybridoma consisted of a 1:1 ratio of antigen and Ia molecules. PMID- 3033073 TI - Phorbol myristate acetate and the calcium ionophore A23187 synergistically induce release of LTB4 by human neutrophils: involvement of protein kinase C activation in regulation of the 5-lipoxygenase pathway. AB - Phorbol myristate acetate (PMA), a tumor-promoting phorbol ester, and the calcium ionophore A23187 synergistically induced the noncytotoxic release of leukotriene B4 (LTB4) and other 5-lipoxygenase products of arachidonic acid metabolism from human neutrophils. Whereas neutrophils incubated with either A23187 (0.4 microM) or PMA (1.6 microM) alone failed to release any 5-lipoxygenase arachidonate products, neutrophils incubated with both stimuli together for 5 min at 37 degrees C released LTB4 as well as 20-COOH-LTB4, 20-OH-LTB4, 5-(S),12-(R)-6-trans LTB4, 5-(S),12-(S)-6-trans-LTB4, and 5-hydroxyeicosatetraenoic acid, as determined by high pressure liquid chromatography. This synergistic response exhibited concentration dependence on both PMA and A23187. PMA induced 5 lipoxygenase product release at a concentration causing a half-maximal effect of approximately 5 nM in the presence of A23187 (0.4 microM). Competition binding experiments showed that PMA inhibited the specific binding of [3H]phorbol dibutyrate ([3H]PDBu) to intact neutrophils with a 50% inhibitory concentration (IC50) of approximately 8 nM. 1-oleoyl-2-acetyl-glycerol (OAG) also acted synergistically with A23187 to induce the release of 5-lipoxygenase products. 4 alpha-phorbol didecanoate (PDD), an inactive phorbol ester, did not affect the amount of lipoxygenase products released in response to A23187 or compete for specific [3H]PDBu binding. PMA and A23187 acted synergistically to increase arachidonate release from neutrophils prelabeled with [3H]arachidonic acid but did not affect the release of the cyclooxygenase product prostaglandin E2. Both PMA and OAG, but not PDD, induced the redistribution of protein kinase C activity from the cytosol to the membrane fraction of neutrophils, a characteristic of protein kinase C activation. Thus, activation of protein kinase C may play a physiologic role in releasing free arachidonate substrate from membrane phospholipids and/or in modulating 5-lipoxygenase activity in stimulated human neutrophils. PMID- 3033074 TI - Human recombinant interleukin 1 beta has no effect on intracellular calcium or on functional responses of human neutrophils. AB - The effect of human recombinant interleukin 1 beta (rIL 1 beta) on human neutrophils was examined. rIL 1 beta, even at concentrations of 100 ng/ml (100 half-maximal T cell stimulating U/ml) did not change significantly the intracellular free calcium concentration, [Ca++]i, whereas the control stimulus, fmet-leu-phe, significantly elevated [Ca++]i. rIL 1 beta also failed to stimulate production of superoxide, degranulation of lysosomal enzymes, phagocytosis of bacterial particles, chemotaxis, or chemokinesis of human neutrophils. This is substantial evidence that superphysiologic concentrations of interleukin 1 have no direct effect on [Ca++]i, as well as on functional responses of neutrophils. PMID- 3033075 TI - The mechanisms of antiviral immunity induced by a vaccinia virus recombinant expressing herpes simplex virus type 1 glycoprotein D: clearance of local infection. AB - We have shown that immunization of mice with a vaccinia virus recombinant expressing glycoprotein D of Herpes simplex virus (HSV)-1 will induce a variety of L3T4+ T cell responses. These included a HSV-specific delayed-type hypersensitivity response, T cell help for the induction of antiviral antibodies, and the ability to eliminate a challenge dose of HSV from the pinna. This protection against a subcutaneous virus challenge was not mediated by the delayed type hypersensitivity response because intravenous inoculation of the vaccinia virus recombinant expressing HSV-1-gD induced a state of split tolerance. Thus, mice could still clear a HSV challenge inoculum from the pinna yet were unable to mount a HSV-specific delayed-type hypersensitivity response. Evidence is presented that suggests the protective response was, at least, in part mediated by a T cell-dependent induction of virus-neutralizing antibodies. Evidence is also presented that may suggest the failure of a vaccinia virus recombinant expressing HSV-1-gD to induce HSV-specific cytotoxic T cell responses appears to minimize the protective response to only efficiently clearing low 10(4) 50% tissue culture infective dose) challenge populations of virus. These findings are discussed with relevance to the immune control of HSV infections and to the future development of anti-HSV vaccines. PMID- 3033076 TI - Structure function relationships for the IL 2 receptor system. III. Tac protein missing amino acids 102 to 173 (exon 4) is unable to bind IL 2: detection of spliced protein after L cell transfection. AB - High affinity receptors for interleukin 2 (IL 2) contain the Tac protein as one ligand-binding subunit. Localization of the IL 2-binding site on this molecule, as well as localization of the complementary site on IL 2, should provide insight into the design of IL 2 analogs. In this report, we examine the ability of normal and modified Tac protein to bind IL 2 and several antibodies that recognize the native Tac molecule. Using a transient L cell expression system, we have determined that transfection with cDNA-missing Tac exon 4 resulted in expression of spliced protein that had no measurable binding to IL 2 or the monoclonal anti receptor antibodies, anti-Tac, and 7G7/B6. This protein was detected, however, by rabbit polyclonal antibodies prepared against synthetic Tac peptides. Thus, one or more amino acids encoded by exon 4 is important, either for direct ligand contact or for the proper folding of critical segments of the Tac molecule. In addition, insertion of stop codons at a unique restriction enzyme site near the beginning of exon 5 resulted in cellular secretion of truncated Tac molecules that were capable of binding IL 2, anti-Tac, and 7G7/B6. Amino acids encoded by exons 5 to 8 thus play no critical role in IL 2 binding. The ligand association demonstrated for truncated Tac protein produced by exons 2 to 4 should guide attempts to define the IL 2-binding segment of the Tac molecule. PMID- 3033077 TI - Thymic epithelium controls thymocyte expression of preleukemic phenotype and leukemogenic retroviruses. AB - Congenitally athymic AKR-streaker (nustr/nustr) mice were grafted separately with syngeneic or allogeneic, irradiated (1200 R) thymic reticuloepithelial (TRE) elements (stroma) or nonirradiated whole thymus grafts (control group) from N tropic murine leukemia virus (MuLV) infection-susceptible (Fv-1n/n) or N-tropic MuLV-infection-resistant (Fv-1b/n) murine strains. From 3 to 13 mo after grafting, the mononuclear cells repopulating the thymus grafts were stained with fluorescent monoclonal antibodies to thymocyte differentiation antigens, peanut agglutinin, and an antibody to MuLV antigens and were then analyzed by flow cytometry. Irradiated TRE of the Fv-1n/n genotype, whether from high or low leukemia-incidence strains, contained lymphoid cells of host (nustr/nustr) origin with alterations in thymocyte differentiation and MuLV antigen expression consistent with preleukemic changes. In contrast, transplanted TRE of the low leukemia-incidence Fv-1b/n genotype restricted preleukemic changes in thymocyte differentiation and MuLV antigen expression by lymphoid cells derived from the nustr/nustr host. Thus, nustr/nustr lymphocytes must infect susceptible TRE (Fv 1n/n) with N-tropic-MuLV before preleukemic changes occur in the mustr/nustr lymphocytes that later migrate to the thymus. Therefore, it was the radiation resistant cells in the thymus that amplified or suppressed expression of AKR MCF retroviruses and the preleukemic phenotype, not the thymic lymphocytes. Thy-1.1+ splenocytes of ungrafted nustr/nustr mice were comparable in percentage to nustr/+ but were deficient in the Lyt-1+2- subpopulation and unresponsive to mitogens or alloantigens in vitro. Analysis of splenocyte cell surface markers, mitogen, MLC, and CML responses of Fv-1n/n-thymus-grafted nustr/nustr mice showed restoration of Lyt-1+2- cells to levels comparable to nustr/+ and reconstitution of in vitro proliferative and cytotoxic responses. PMID- 3033078 TI - Cell surface antigens on rat islet tumors. AB - To examine the antigenic properties of the rat pancreatic beta cell tumor, RIN, we used a cell surface enzyme-linked immunosorbent assay (CELISA). Monoclonal antibodies of known specificity are used to validate the assay, and the results show that antibodies directed at glycoproteins and glycolipids are detected by this technique. The principal glycolipid targets are found in the ganglio and lacto series of glycosphingolipids, and not in the globo series. When the CELISA was used to study sera from type I diabetics, no differences in binding of control and diabetic samples were detected at low dilutions of serum. However, at higher serum dilutions (1/30 to 1/120) binding of IgG antibodies from type I diabetics was greater than that observed with normal sera. Similar reactivity was not detected in these sera when rat hepatocytes were used as targets in the CELISA. Prolonged culture of RIN cells, however, resulted in the loss of reactivity with sera from type I subjects, and is associated with a corresponding decrease in expression of cell surface gangliosides. Accordingly, subclones selected for ganglioside expression were used to compare anti-RIN binding in type I diabetics with that of normal controls and unaffected siblings. The results indicate that some rat islet tumors express antigenic determinants recognized by anti-islet antibodies associated with type I diabetes. Both nonspecific interaction at low serum dilutions and variable expression of cell surface antigens may explain the difficulties encountered when these cells are used for diagnostic purposes. An objective assay such as the CELISA may help to avoid these problems. PMID- 3033080 TI - The immune status of avian sarcoma virus-infected chickens as a determinant of sarcoma growth pattern and viral antigen expression. AB - Patterns of sarcoma growth were compared in immune-suppressed (cyclophosphamide treated) vs nonsuppressed avian sarcoma virus-infected chickens. Sarcomas were initially induced in suppressed or nonsuppressed line FP chickens (donors) by wing web inoculation with clone 85, an avian sarcoma virus encoding an envelope glycoprotein that is non-antigenically cross-reactive with endogenous viral glycoprotein. Sarcoma tissue from these donors was then implanted in the wing webs of suppressed or nonsuppressed recipients to compare the effects of immune suppression of donors and of recipients. Sarcoma tissue that had been excised from suppressed donors and implanted in the wing webs of nonsuppressed recipients evidenced a much greater capacity than sarcoma tissue from nonsuppressed donors to yield distal sarcomas localized to visceral organs and to induce expansion by infection-mediated recruitment of new sarcoma cells. In contrast, immune suppression of the recipients of sarcoma tissue from nonsuppressed donors was not significantly enhancing for these effects. The enhancement achieved by immune suppression of the donors correlated with a markedly increased level of virus production and viral antigen expression by the primary (wing web) sarcoma cells from these donors. PMID- 3033081 TI - CSF-1-induced resistance to viral infection in murine macrophages. AB - Murine peritoneal thioglycollate-elicited macrophages were cultured for 3 days in the presence or absence of highly purified human macrophage colony stimulating factor (CSF-1). The cells were then challenged with vesicular stomatitis virus (VSV) for 24 hr. Ability to resist viral infection was measured in two ways. First, macrophage viability after infection with VSV was measured by washing to remove dead cells, staining the remaining cells with crystal violet, and reading absorbance. Second, a yield reduction assay was used to measure viral replication in the macrophage cultures. Cells treated with CSF-1 (500 to 2000 U/ml) and infected with VSV looked similar microscopically to uninfected cells and had absorbance values twofold to threefold higher than those of infected cultures not treated with CSF-1. The CSF-1-treated cultures also had a virus titer one log lower than that of the untreated cultures. Treatment with partially purified murine CSF-1 induced a similar reduction in virus titer, whereas other murine CSF tested (purified murine GM-CSF, lung-conditioned medium that contains GM-CSF and G-CSF, and WEHI-3B-conditioned medium as a source of IL 3) had little to no effect on virus titer. Antibody to murine IFN-alpha/beta added to the macrophage cultures inhibited the protective effect of CSF-1, indicating that the CSF-1 effect was due to induction of endogenous IFN. Treatment with lipopolysaccharide (1 ng/ml) had some protective effect, which was blocked with polymyxin B. Polymyxin B did not inhibit the effect of CSF-1. PMID- 3033079 TI - Purification and biochemical characterization of a human autocrine growth factor produced by Epstein-Barr virus-transformed B cells. AB - Autocrine growth factor for Epstein-Barr virus-transformed human B cells (aBGF), a protein that is constitutively produced by the human EBV-transformed B cell line 5/2, has been purified from serum-free conditioned medium. The purification involved sequential ammonium sulfate precipitation, ion exchange chromatography, gel filtration, and reversed-phase high performance liquid chromatography. The purified protein has a m.w. of 16,000 in NaDodSO4/polyacrylamide gel electrophoresis and an isoelectric point between 7.0 and 8.0. The relative molecular mass 16,000 form exists in equilibrium with dimeric and tetrameric forms. aBGF supports the growth of EBV-transformed B cells, which have been deprived of their own conditioned medium. The purified aBGF is fully effective at 0.5 ng/ml and has no interleukin 1 activity in the lymphocyte activation factor assay. Because several randomly selected lines of EBV-transformed cells and one EBV-negative lymphoma cell line both produce aBGF activity and show growth dependency on aBGF and because stimulation of normal B cells with anti immunoglobulin M is increased by aBGF, we propose that aBGF has general significance for growth control of human B cells. PMID- 3033082 TI - Excess antibody immunoassay for the measurement of tonin in rat tissues and plasma. AB - Tonin, a proteolytic enzyme isolated from the rat submandibular gland, can generate angiotensin II directly from angiotensinogen. To date a method for the measurement of tonin in plasma has not been available and the present paper describes a sensitive and specific excess antibody immunoassay for determination of tonin in tissue homogenates and plasma. Interference from immunologically cross-reacting proteins was evaluated and the assay was found to be specific for tonin. Tonin measured in various tissue homogenates was directly proportional to the amount of sample added, giving a linear dose-response curve. The slope of this curve was determined by the recovery of tonin, which was better than 55% for urine and all tissues tested. The highest concentration of tonin was seen in the submandibular and sublingual gland (69 and 0.7 microgram/mg protein, respectively). The parotid gland, the exorbital lacrimal gland, liver, kidney, pancreas, and lung contained only negligible amounts (less than 4 ng/mg protein). Tonin in plasma was bound to one major inhibitor with a molecular weight of about 650,000-750,000. A partial splitting of the tonin-inhibitor complex was obtained by preincubating plasma with guanidine, allowing tonin to be measured with a recovery of 38 +/- 13% (n = 16) and with a linear dose-response curve. The concentration of immunoreactive tonin in normal arterial plasma from adult male rats was 0.90 +/- 0.53 ng/ml (n = 16). The concentration decreased after removal of the submandibular glands and increased after sympathetic stimulation. PMID- 3033083 TI - More exact quantification of interleukin-2 production by addition of anti-Tac monoclonal antibody to cultures of stimulated lymphocytes. AB - Human T-cells produce interleukin-2 (IL-2) in response to in vitro stimulation with mitogens and antigens. Precise measurement of IL-2 production in vitro is hampered by binding of IL-2 to IL-2 receptors on activated T-cells which results in IL-2 consumption. In an attempt to prevent IL-2 consumption, we supplemented the cultures with anti-Tac, a monoclonal antibody that functionally blocks the human lymphocyte receptor for IL-2. Addition of anti-Tac to cultures of mitogen stimulated peripheral blood mononuclear cells resulted in a 2-4-fold increase of maximal levels of IL-2 in the supernatants. No decline of IL-2 concentrations beyond 18-24 h of culture was observed, indicating that anti-Tac effectively blocked IL-2 consumption. Comparison of the kinetics of IL-2 accumulation in the presence and absence of anti-Tac revealed that IL-2 consumption by mitogen stimulated cells occurred as early as 6 h after culture initiation. Addition of anti-Tac to cultures of antigen-stimulated cells similarly resulted in higher IL 2 levels in the culture supernatants, and prevented the rapid decline of IL-2 concentrations at prolonged culture time. We conclude that addition of anti-Tac to lymphocyte cultures enables more exact quantification of in vitro IL-2 production, presumably by blocking IL-2 consumption. This finding offers new possibilities for the study of abnormalities of IL-2 production in different disease conditions. PMID- 3033085 TI - Krukenberg tumour associated with pregnancy. PMID- 3033084 TI - Long-term storage of peroxidase-labelled immunoglobulins for use in enzyme immunoassay. AB - Optimal conditions for long-term storage of peroxidase-labelled immunoglobulins for use in enzyme immunoassays have been investigated with particular emphasis on the preservation of the immunological and enzyme activity. Immunoglobulin G preparations from different animal species were purified according to conventional methods and labelled with horseradish peroxidase. Conjugates were stored at different temperatures either in liquid form with 50% glycerol, after lyophilization or as ammonium sulphate precipitates. In addition, the conjugates were also frozen and stored at -40 degrees C. The immunological and enzymatic activity of the conjugates was evaluated at regular intervals throughout storage. The best results were obtained with conjugates stored as ammonium sulphate precipitates at 4 degrees C. Under these conditions, the complexes retained after 2 years of storage 92 and 91% of their enzymatic and immunological activity respectively and gave excellent reproducibility in ELISA. The conjugates exhibited a very low degree of self-aggregation. The ammonium sulphate did not interfere in the ELISA. Storage at elevated temperatures resulted in a greater decrease in enzymatic than immunological activity but the results obtained with the ammonium sulphate precipitates were clearly superior to those obtained with alternative storage procedures. The estimated half life of the whole enzymatic immunological activity of conjugates at 4 degrees C was 9 years. The stability of both activities at 25 degrees C was also reasonably good for short periods. Conjugates stored for 40 days at 37 degrees C lost 60% enzyme activity but retained unaltered their immunological activity and gave rise to high blanks in the ELISA, due to auto-aggregation. PMID- 3033086 TI - Serum folate in viral and mycoplasmal infections. AB - The concentration of serum folate in 260 patients with viral and mycoplasmal infections was determined (305 samples). In 60% the serum folate concentrations were found to be below 3 micrograms/l. The incidence of low serum folate varied slightly. According to the infections diagnosed, low values were: for influenza A 50% (50 patients), influenza B 42% (45 patients), human parvovirus 67% (76 patients), rubella 62% (13 patients), infectious mononucleosis 60% (15 patients), Mycoplasma pneumoniae 73% (45 patients). For a group of undiagnosed rashes it was 81% (16 patients). The cause of low concentrations of serum folate in these patients is discussed. PMID- 3033087 TI - Plasma membrane receptors. PMID- 3033088 TI - In vitro damage of basal lamina-associated anionic sites by hydroxyl radical. AB - We examined the effect of chemically generated hydroxyl radical on basal lamina associated anionic sites. Cytochemical studies showed that hydroxyl radical produced a loss of cationic tracer-positive, anionic particles, and this effect was inhibited by a specific scavenger, thiourea. These data might suggest that anionic sites were degraded by hydroxyl radical which was derived, for example, from the activated leukocytes in close contact with the dermal-epidermal junction during acute inflammation resulting in the disturbance of the charge-selective permeability barrier. PMID- 3033089 TI - Molecular epidemiology of adenovirus type 35 infections in immunocompromised hosts. AB - Adenovirus type 35 (Ad35) is an uncommon group B adenovirus that has been isolated nearly exclusively from immunosuppressed hosts. In our series of 46 patients with Ad35 isolates, 36 had AIDS or AIDS-related complex (ARC), seven patients were immunocompromised because of other diseases, and three patients were "normal." Neither patients with AIDS or ARC made specific antibody to Ad35, whereas antibody was present in one immunocompetent host and one renal transplant recipient. All isolates were identified as Ad35 by using genomic analysis with the restriction endonuclease SmaI, but many viruses exhibited other group B hemagglutinins, a property of the fiber protein. Further analysis of DNAs from 40 of these isolates, mapped by using the enzymes BamHI, HpaI, and PstI, revealed a total of 14 genomic variants from the Ad35 prototype. Seven of these genomic types, with either Ad7 or Ad11 hemagglutinin, had corresponding restriction site differences within the fiber gene, a result consistent with recombination events with other group B adenoviruses. PMID- 3033090 TI - Direct serotyping of human rotavirus in stools by an enzyme-linked immunosorbent assay using serotype 1-, 2-, 3-, and 4-specific monoclonal antibodies to VP7. AB - Four serotypes of human rotaviruses that can be differentiated by neutralization tests have been described so far. We prepared serotype 1-, 2-, 3-, and 4 specific, neutralizing monoclonal antibodies to human rotaviruses. All were directed to VP7, a glycoprotein that carries a major serotype specificity. An enzyme-linked immunosorbent assay using these monoclonal antibodies has been developed for serotyping human rotavirus isolates. The sensitivity and specificity of this method were verified by using various strains of human rotavirus that were adapted to cell culture. Furthermore, it was shown that the method was applicable to serotyping human rotaviruses directly in stool specimens. PMID- 3033091 TI - Antigen detection with monoclonal antibodies for the diagnosis of adenovirus gastroenteritis. AB - A monoclonal antibody-based enzyme immunoassay (EIA) was developed for direct detection of enteric adenoviruses in stool specimens from individuals with gastroenteritis. Tests specific for each of the enteric adenoviruses, adenovirus type 40 (Ad 40) and type 41 (Ad 41), were designed. The sensitivity of the assay was determined by comparing the results of the EIA with isolation of virus in Graham 293 cells from stools that contained particles having adenovirus morphology. The standard for specificity was analysis of adenovirus genome profiles after digestion with SmaI endonuclease. The sensitivity was 95.8% (23 of 24) for Ad 40 and 97.1% (34 of 35) for Ad 41. The specificity was 95.7% (45 of 47) and 97.2% (35 of 36), respectively. The two type-specific monoclonal antibodies could be mixed in an EIA for identification of enteric adenoviruses in stools without loss of reactivity in either type. The EIA permits rapid diagnosis and type-specific identification of enteric adenoviruses in gastroenteritis. PMID- 3033092 TI - Detection of antigen to herpes simplex virus in cerebrospinal fluid from patients with herpes simplex encephalitis. AB - Cerebrospinal fluid (CSF) specimens were obtained from patients with presumed herpes simplex encephalitis who underwent brain biopsy for diagnostic confirmation. Coded CSF specimens were fixed on nitrocellulose filter paper and probed with a pool of monoclonal antibodies directed against four herpes simplex virus glycoproteins (gB, gC, gD, and gE). Herpes simplex virus antigen was detected in 35 of 40 specimens obtained from 26 biopsy-positive patients. In contrast, only three of 25 specimens from 17 biopsy-negative patients gave positive results by this assay. An additional 30 CSF specimens from patients with proven bacterial and fungal infections were all negative by this assay. For all specimens tested, the sensitivity and specificity of the assay were both 88%. However, when results were evaluated by patient, the sensitivity was 92% (24 of 26) with a specificity of 82% (14 of 17). Among specimens collected one week or later after disease onset, the sensitivity was 100%, with a specificity of 93%. PMID- 3033093 TI - Defective production of antibody to herpes simplex virus in neonates: defective production of T helper lymphokine and induction of suppression. AB - Defective production of antibodies to herpes simplex virus (HSV) and low resistance to HSV infection in neonatal mice could be reconstituted by using macrophages from syngeneic adult mice plus concanavalin A-stimulated supernatants prepared from adult mouse spleen cells or adult human peripheral blood lymphocytes. In each supernatant, interleukin-2 (IL-2) produced by T helper cells (positive for Lyt 1.2 or OKT4) was necessary to provide reconstitution. Supernatants from neonatal mice failed to mediate reconstitution because of an age-dependent absence of production of IL-2. Although supernatants from neonatal human cell cultures contained IL-2, they failed to reconstitute production of antibody to HSV and resistance to HSV infection because of a suppressor of IL-2 activity. Early antibody production and antibody-dependent cellular effector function are important defenses against HSV infection and are critically defective in the neonate. PMID- 3033094 TI - Protection of guinea pigs from primary and recurrent herpes simplex virus (HSV) type 2 cutaneous disease with vaccinia virus recombinants expressing HSV glycoprotein D. AB - Vaccinia virus recombinants containing herpes simplex virus (HSV) type 1 (VP176) or type 2 (VP221) glycoprotein D (gD) genes were studied for their protective potential in the guinea pig model of recurrent HSV type 2 disease. Cells infected with these recombinants synthesized at least one protein (precursor, mature form, or both) that was precipitated with monoclonal antibody to HSV type-common determinants on gD. These determinants were detected on the surface of cells infected with the recombinants at 2 hr after infection. VP176 immunization protected against primary (P much less than .001) and recurrent (P much less than .001) cutaneous HSV type 2 lesions and ganglionic latency (62% protection). VP221 immunization protected against recurrent disease (P less than .05), although HSV type 2 ganglionic infection was established. Protection, first observed at two weeks after immunization, apparently did not involve HSV-specific neutralizing antibody because seroconversion was detected at 35-45 days after immunization. Protection was correlated with HSV-specific lymphoproliferation and the elaboration of lymphokines that enhance natural killer cell cytolysis. PMID- 3033095 TI - Outer membrane protein F (porin) preparation of Pseudomonas aeruginosa as a protective vaccine against heterologous immunotype strains in a burned mouse model. AB - Outer membrane (OM) protein F (porin) was purified by extraction from polyacrylamide gels of cell envelope proteins of the Pseudomonas aeruginosa PA01 strain. Mice were immunized intramuscularly with 10 micrograms of protein F preparation on days 1 and 14 and then subjected to burn and challenge on day 28. Protein F immunization afforded significant protection above that provided by PA01 lipopolysaccharide (LPS) immunization against subsequent challenge with six of six heterologous LPS immunotype strains of P. aeruginosa. By an ELISA, the murine immune response revealed an IgG titer of 5,120 to protein F by day 30. Immunoblot analysis of antisera from protein F-immunized mice revealed bands with both protein F and protein H of cell envelopes of all immunotypes tested. Active immunization with OM protein H did not, however, afford significant protection to mice in this burned mouse model. These data show the efficacy of OM protein F as a protective vaccine in a murine model representative of human infection. PMID- 3033096 TI - Detection of human papillomavirus type 16 DNA in peritoneal washings from patients with cervical carcinoma. PMID- 3033097 TI - Failure of live, oral virus vaccines in developing countries. PMID- 3033098 TI - [Signals for T cell growth]. PMID- 3033099 TI - [A self protection system based on the oxidation, oxygenation and halogenation by myeloperoxidase]. PMID- 3033100 TI - NDA3: a differentiation antigen associated with the receptor for B cell growth factor. PMID- 3033102 TI - [Radiotherapy of patients with germ cell tumor]. PMID- 3033103 TI - [When does chemotherapy of trophoblastic disease should be stopped?]. PMID- 3033101 TI - Regulation of T cell activation by leukotriene B4. PMID- 3033105 TI - Eccrine tumours of the hand: two cases. AB - Two cases of eccrine tumour of sweat gland origin found in the hand are presented. They were non-tender rounded masses of eight months to two years duration, beneath and adherent to the epidermis but movable on underlying tissue. The pre-operative diagnosis in each case was that of a sebaceous cyst. Histology revealed one to be an eccrine spiradenoma and the other a chondroid syringoma. The lesions were considered unusual in that such tumours usually occur on the trunk, head and back and are rarely found in the hand. PMID- 3033104 TI - [A case of a simple virilizing form of 21-hydroxylase deficiency with schizophrenic symptoms, marked pigmentation and highly elevated plasma deoxycorticosterone]. PMID- 3033106 TI - Respiratory burst of normal human eosinophils. AB - Oxidative metabolism of eosinophils has generally been studied in cells from patients with eosinophilia. We isolated eosinophils with purity of greater than 95% from normal donors. Oxygen metabolism of eosinophils and neutrophils was compared using O2-. production as a measure of the stimulus-induced respiratory burst. During 10 min of stimulation, using phorbol myristate acetate (PMA) or opsonized zymosan (OPZ), eosinophils produced two to three times as much O2-. as did neutrophils. Continuous kinetic analysis was used to measure the latency period and maximal rate of O2-. production. Eosinophils has a shorter latency period and greater rate of O2-. production than did neutrophils at all concentrations of PMA. With OPZ stimulation, O2-. production rates by both cells were similar. The latency periods were similar with high concentrations of OPZ but at limiting amounts of OPZ eosinophils had a longer latency period than did neutrophils. In addition, the kinetic assay demonstrated that the respiratory burst in eosinophils was more sustained than in neutrophils. These results indicate substantial differences in the oxidative burst of eosinophils and neutrophils with respect to activation, capacity, and regulation. These distinctive features of O2-. production by eosinophils may be important in host defense against metazoan parasites or in tissue injury during inflammation. PMID- 3033107 TI - Candidacidal activity of monocyte-derived human macrophages: relationship between Candida killing and oxygen radical generation by human macrophages. AB - Freshly isolated human monocytes ingested and killed Candida albicans, and generated O2- H2O2 and .OH efficiently. When monocytes were cultured in vitro, these cells transformed into macrophages. Cultured monocytes retained their ingestive activity but lost their candidacidal activity almost completely after day 3. The release of O2- by monocytes decreased slightly with culture and that of .OH was markedly decreased on day 3 of culture. The activity of myeloperoxidase in the monocytes decreased with culture. These results suggested that the loss of candidacidal activity is due to the decrease of .OH generation and myeloperoxidase activity in cultured monocytes. PMID- 3033108 TI - Independent regulation of leukotriene B4-elicited polymorphonuclear leukocyte exocytosis and superoxide generation by a serum factor. AB - Serum from a patient with inactive systemic lupus erythematosus (SLE) and ibuprofen-induced transient neutropenia was used as a probe to define further the control of human polymorphonuclear leukocyte (PMN) exocytosis and superoxide (O2 ) generation. Thirty-minute preincubation of normal PMNs with 10-50% v/v of this serum, followed by washing, produced a specific dose-related suppression of leukotriene B4 (LTB4)-elicited beta-glucuronidase and lysozyme release of up to 45% and 30% respectively. If cells were not washed, the inhibition increased to 60% and 40%. Superoxide production stimulated by LTB4 was unaffected. The serum had no effect on formyl-met-leu-phe (FMLP) or phorbol myristate acetate stimulated O2- or exocytosis. O2- and beta-glucuronidase release elicited by zymosan-treated serum (ZTS) were both decreased by 15%, but there was no increased inhibition seen if cells were not washed, or if the time of preincubation was increased from 7 to 30 min. In contradistinction, the serum inhibition of LTB4 exocytosis did show time dependence. Serum obtained when the patient was not leukopenic and sera from 6 normal controls, 2 patients with inactive SLE, 1 patient with SLE and chronic leukopenia, and 2 controls taking ibuprofen did not influence any PMN function. The serum inhibition of ZTS-induced functions was qualitatively similar to that observed when PMNs were preincubated and desensitized with ZTS in vitro. Selective inhibition of LTB4 exocytosis was not seen when PMNs were desensitized with LTB4 in vitro. These observations indicate that LTB4-elicited O2- and exocytosis can be independently and specifically regulated. The cellular site at which this serum factor acts is not clear, but the current studies strongly suggest that this inhibition is not due to in vitro deactivation by LTB4 activity. PMID- 3033109 TI - Nitroblue tetrazolium reduction in lymphocytes. AB - Certain populations of lymphocytes have been shown to reduce tetrazolium salts, indicating that superoxide anion may be present in lymphocytes. These experiments were done to determine if nitroblue tetrazolium reduction by lymphocytes was due to the presence of superoxide anion. Mitogen-activated lymphocytes showed increased nitroblue tetrazolium reduction compared to unstimulated cells, and the cells reducing nitroblue tetrazolium were both T-cells and non-T-cells. Release of superoxide anion or hydrogen peroxide by either resting or stimulated lymphocytes was not detected. There was no difference between resting and stimulated lymphocytes in the amount of chemiluminescence produced in the presence of lucigenin, an agent which appears to be sensitive to intracellular and extracellular superoxide anion. The results, then, indicate superoxide anion is not present in lymphocytes. PMID- 3033110 TI - Detection of the production of oxygen-centered free radicals by human neutrophils using spin trapping techniques: a critical perspective. AB - Human neutrophils generate oxygen reduction products as a consequence of membrane interactions with a number of stimuli. One oxygen-centered free radical (superoxide) has been unequivocally shown to result from this "respiratory burst," and some experimental evidence for another (hydroxyl radical) has been published, although debate remains as to its significance. The role of phagocyte derived free radicals in microbicidal and tumoricidal activity as well as tissue damage at sites of inflammation has been the focus of extensive investigation in recent years. Of the techniques available to study free radical generation in biological systems, spin trapping has emerged as a powerful tool for detection and identification of these reactive species, owing in part to its ability to measure production of free radicals inside the phagosome. However, interpretation of resulting spectra is extremely complex and filled with pitfalls and limitations. In this communication we review spin-trapping techniques and discuss the application of this system to the identification of free radicals resulting from stimulation of human neutrophils. Criteria are established which are necessary for definitive identification of superoxide and hydroxyl radical when employing this technology. In this context a critical perspective of previous studies of neutrophil-derived free radicals is offered. PMID- 3033111 TI - Late aluminum therapy reduces the cellular activities of simple silicosis in the sheep model. AB - To evaluate the biological effects of aluminum lactate therapy on nodular silicosis, we exposed the tracheal lobe of three groups of sheep containing eight sheep per group to either 11 mg of Al lactate in 100 ml saline (Al group), 100 mg of Minusil-5 in 100 ml saline (Si group), or 100 mg of Minusil-5 in 100 ml saline followed by 11 mg of Al lactate at monthly intervals 4 months after exposure (Si Al-treated group). The lung biological processes were evaluated by sequential lung lavage analyses of cellularity and biochemistry of supernatant and by autopsy analyses of cellularity and biochemistry of supernatant and by autopsy analyses of lung tissue histopathology and quartz content. Al lactate alone did not have any significant effect. Silica exposure produced the silicotic nodules and significant increases on lung lavage of cellularity, enzyme release, surfactant, and glycosaminoglycan accumulations. Al lactate therapy at month 4 after exposure did not decrease the pathological score of disease, but it significantly reduced all markers of cellular hyperactivity. This therapy was associated with a 65% reduction of the quartz retention in lung tissue and might help to prevent long-term progression of the disease process. PMID- 3033113 TI - Requirements for immunoglobulin synthesis in leukocyte cultures exposed to human cytomegalovirus. AB - T cell-depleted leukocytes from normal healthy donors lacking antibody to cytomegalovirus (CMV) were induced to secrete immunoglobulin (Ig) by exposure to inactivated CMV. The responses to two virus strains were compared. One strain had been reported to require specific T cell help to induce Ig synthesis, and the other had been reported to induce Ig synthesis in the presence of only very few cells. Both viruses induced T cell-depleted leukocytes from one group of seronegative donors to secrete Ig. However, both viruses failed to induce leukocytes from a second group of donors to secrete Ig unless B cell growth factor was added as a source of T cell help. These findings suggest that certain individuals are hyperresponsive to CMV and raise the possibility that this group may be most likely to develop mononucleosis after primary infection with CMV. PMID- 3033114 TI - The effect of a protein kinase C inhibitor, H-7, on human neutrophil oxidative burst and degranulation. AB - The role of C-kinase in the activation of human polymorphonuclear leukocytes has been examined using H-7, a recently described C-kinase inhibitor. We found that H 7 will inhibit PMN superoxide anion release in response to the tumor promotor phorbol myristate acetate and the calcium ionophore A23187. In contrast, no inhibition by H-7 was seen for PMN superoxide release stimulated by the chemotactic peptide FMLP. H-7 did not inhibit PMN NADPH oxidase activity or PMN degranulation by any stimulant, but it reversed a phorbol ester-induced inhibition of granule release by FMLP. The results show that H-7 differentially affects the PMN functional events of secretion and superoxide release and suggests that an H-7 inhibitable C-kinase is not involved in chemotactic peptide induced activation of PMN and may not regulate stimulus induced PMN degranulation. PMID- 3033112 TI - Alveolar macrophage phagocytic kinetics following pulmonary parainfluenza-3 virus infection. AB - Qualitative and quantitative evaluations of the cellular components of bronchoalveolar washings of calves with experimental parainfluenza-3 virus pneumonitis and control calves were made. Calves were exposed to 10(9) TCID50 of PI-3 by intranasal aerosol exposure and bronchoalveolar cells obtained 7 days after infection by volume-controlled bronchopulmonary lavage. Transient tachypnea and pyrexia occurred in all infected calves, and virus was recoverable at 7 days from nasal swabs and lung tissue. Pulmonary lesions were typical of viral pneumonitis, characterized by patchy alveolitis and bronchiolitis with accumulations of cells and inflammatory debris. The mean total lavage cell yield was elevated in the virus-infected calves, and the percentage of neutrophils was elevated (P less than 0.05). Increased numbers of pulmonary alveolar macrophages (PAM) were also recovered but the difference was not significant. Linear regression equations showed that a decreased proportion of PAM from virus infected animals were phagocytic. The mean initial phagocytic rates of macrophages from calves with viral pneumonitis were significantly decreased (P less than 0.05) over controls. This difference was concentration dependent and required a phagocytic stimulus in excess of 12.5 X 10(6) beads/ml. Studies of phagocytic kinetics showed that PAM from calves with viral pneumonitis had a lower Vmax than PAM from control calves, but that Km values were comparable. No differences in PAM beta-glucuronidase and acid phosphatase activity were observed. These results indicate depressed phagocytic function in PI-3 virus inflamed lungs relative to controls. In concert with virus-induced airway lesions, such in vivo depression of PAM phagocytic functions would be expected to depress pulmonary particulate clearance and lung defense mechanisms. PMID- 3033115 TI - Zymosan-stimulated production of phosphatidic acid by macrophages: relationship to release of superoxide anion and inhibition by agents that increase intracellular cyclic AMP. AB - Murine peritoneal macrophages (m phi) respond to unopsonized zymosan with the production of superoxide anion (O2-). We investigated the involvement of phospholipid turnover in the transduction mechanism for this phenomenon. Zymosan stimulated m phi produced increased amounts of phosphatidic acid (PA); the increase was first detected at 1.5 min and continued for 10 min of incubation. Production of O2- was not detected until between 2 to 4 min after stimulation, and continued to increase through 60 min. Inhibition experiments suggested that these two processes were linked. Theophylline (theo)/dibutyrylcyclic AMP (dbcAMP) and theo/prostaglandin E2 (PGE2) inhibited O2- production at every time point (79% and 80% inhibition, respectively, at 4 min). Corresponding inhibition of PA production was also achieved at every time point (85% by theo/dbcAMP; 67% by theo/PGE2 at 4 min). These results are compatible with a role for phospholipid remodeling in the transduction process associated with the respiratory burst. Results suggest that the phospholipid species could be phosphatidylcholine (PC) as well as phosphatidylinositol (PI). PMID- 3033116 TI - Characterization of the phospholipid and fatty acid composition of Sendai virus. AB - The lipid composition of Sendai virus, propagated in chicken eggs, was analyzed by high performance liquid chromatography (HPLC), thin-layer chromatography (TLC), and gas-liquid chromatography (GLC). Phosphatidylcholine was found to be the dominant phospholipid (37.3%) with phosphatidylethanolamine (26.8%) and phosphatidylserine (12.0%) also present in significant amounts. Analysis of the fatty acid methyl esters revealed that the dominant fatty acids in total phospholipid were: C16:0 (17.6%), C18:0 (15.4%), C18:1 (n-9) (22.0%), and C24:0 (6.0%). Cardiolipin, phosphatidylserine, and sphingomyelin contained higher levels of saturated fatty acids relative to phosphatidylinositol, phosphatidylethanolamine, and phosphatidylcholine. PMID- 3033117 TI - Sidedness of ceramide-phosphoethanolamine synthesis on rat liver and brain microsomal membranes. AB - Phosphatidylethanolamine:ceramide-ethanolamine-phosphotransferase catalyzes the synthesis of ceramide-phosphoethanolamine, a sphingomyelin analogue. Its localization was studied in rat liver and brain microsomes. After testing the integrity and the sidedness of microsomal vesicles, trypsin treatment of intact or deoxycholate-disrupted microsomes made it possible to conclude that both the transferase and the ceramide-phosphoethanolamine are located in the cisternal leaflet of the membrane bilayer. Using trinitrobenzenesulfonic acid as a probe, no trace of newly synthesized ceramide-phosphoethanolamine was detectable on the cytoplasmic side of the microsomes. PMID- 3033118 TI - Chylomicron remnant-vitamin A metabolism by the human hepatoma cell line HepG2. AB - The binding and metabolism of [3H]vitamin A-containing chylomicron (CM) remnants by the human hepatoma cell line HepG2 were studied. Mesenteric lymph chylomicrons were collected from [3H]retinol-fed rats and incubated with lipoprotein lipase to obtain CM remnants. At 4 degrees C, specific CM remnant binding was inhibited by an excess of unlabeled CM remnants. Specific binding predominated at low concentrations and approached saturation while total binding continued to increase over an extensive concentration range (0.45-32 microgram triglyceride/ml). CM remnant uptake at 37 degrees C was greater than that of CM and at least 70 times more efficient than the pinocytosis of sucrose. CM remnant binding increased with the extent of lipolysis. Addition of human apolipoprotein E enhanced both CM remnant and CM binding. After internalization, HepG2 cells hydrolyzed CM remnant-[3H]retinyl esters, and radiolabeled metabolites accumulated. As a function of the concentration of [3H]retinoid initially bound to cells, retinol and retinyl esters accumulated as the major cell-associated metabolites. In contrast, retinol was the major metabolite in the medium only at low retinoid concentrations; other more polar metabolites accumulated at higher concentrations (greater than 110 pmol retinoid/mg cell protein). The accumulation in the medium of labeled metabolites derived from CM remnant-retinoid was reduced when cells were preincubated in unlabeled retinol-supplemented media. The specific activity of retinol in the medium indicated that CM remnant-vitamin A had mixed with the cellular store prior to its secretion as retinol. These results indicate that HepG2 cells internalize CM remnants in part by specific binding sites, and that the metabolism of CM remnant-retinoids by the HepG2 cell involves retinyl ester hydrolysis and the secretion of retinol and other more polar metabolites. These processes were regulated in part by the concentration of retinoid delivered by the CM remnant and by the initial retinoid content of the cell. PMID- 3033119 TI - Prevention of traveler illness. PMID- 3033120 TI - Analysis of dihydrocodeine in urine using Sep-Pak C18 cartridges for sample cleanup. AB - A rapid and precise method for the isolation and identification of dihydrocodeine from urine is reported. The narcotic is isolated from urine using Sep-Pak C18 cartridges for cleanup, requiring less than 30 min for preparation. Identification is performed by gas chromatography/mass spectrometry. PMID- 3033121 TI - The physico-chemical, immunological and biological characteristics of HCG in gestational trophoblastic disease and pre-eclampsia. PMID- 3033122 TI - N-hexane-induced electroneurographic changes and early detection of N-hexane intoxication. PMID- 3033123 TI - Nucleotide sequence of the coding and flanking regions of the human parainfluenza virus type 3 fusion glycoprotein gene. AB - The complete nucleotide sequence of the human parainfluenza virus type 3 (HPIV3) fusion (F) protein gene has been determined. The HPIV3 F gene is 1851 nucleotides long including six U residues in the genomic RNA, which probably direct synthesis of the first few nucleotides in the F mRNA polyadenylate tail. The HPIV3 F gene contains a single long open reading frame coding for 539 amino acids. The predicted molecular weight of the unglycosylated precursor F0 protein was 60031. Four potential carbohydrate acceptor sites were identified. Comparison of the HPIV3 F protein sequence with the F gene sequences of two other paramyxoviruses, Sendai virus and simian virus 5, indicated a very close evolutionary relationship between HPIV3 and Sendai virus. Sequence analysis of HPIV3 F gene flanking regions identified signals which appear to be responsible for polymerase recognition and polyadenylation. PMID- 3033124 TI - Lymphotropic strain SL3 of Aleutian disease virus: identification of replicative form DNA, molecular cloning and expression of capsid-specific proteins. AB - Replicative form (RF) DNA of the lymphotropic strain SL3 of Aleutian disease virus was isolated from infected cell cultures. A novel intermediate of about 7.6 kilobases was demonstrated in Hirt lysates in addition to single-stranded viral, double-stranded monomer and dimer RF DNA. The monomer RF DNA exhibited a length heterogeneity of 70 bp and 160 bp at its 3' and 5' termini. The two major monomer RF DNA species each contained hairpins in the extended or the foldback configurations. A central fragment between map units 0.15 and 0.88 was cloned into plasmid pUC18. The recombinant clone expressed virus-specific proteins ranging from 32,000 to 74,000 mol. wt. PMID- 3033125 TI - Organization of terminal reiterations in the virion DNA of herpesvirus saimiri. AB - The population of herpesvirus saimiri (HVS) genomes extracted from extracellular virions are double-stranded, linear DNA molecules of about 160 kilobase pairs (kbp) each composed of a central segment of 110 to 112 kbp and 36% (G + C) (i.e. 'light' or L-DNA) linked to direct reiterations of a 1.44 kbp repeat unit of 71% (G + C) (i.e. 'heavy' or H-DNA) at each terminus. In this paper, we show that the population of HVS DNA molecules contains approximately equal concentrations of genomes with all possible integral numbers of complete repeat units (i.e. from greater than 30 to 1) at either 'left' or 'right' ends but that all molecular ends are derived by a unique cleavage at a site close to the single ApaI restriction endonuclease site of the H-DNA repeat unit. Junctions of proximal H DNA repeat units with L-DNA occur at, or very close to, the sequence present at the molecular ends. The transition from L- to H-DNA occurs abruptly at this site at the 'right' end of the L-DNA component but some rearranged restriction enzyme cleavage sites typical of H-DNA are found within the first 0.8 kbp of the L-DNA sequences at the 'left' H-L DNA junction. HVS appears to provide an extreme example of the general process whereby herpesvirus DNAs are matured from concatemeric intermediates by a site-specific cleavage/recombination process involving random choice between equivalent sites for the initiation of the process and with choices between alternative termination sites being limited by a headful packaging mechanism. PMID- 3033126 TI - Genetic relations between varicella-zoster virus and Epstein-Barr virus. AB - Varicella-zoster virus (VZV) and Epstein-Barr virus (EBV) are important human pathogens which belong to different subfamilies of the herpesviruses: the Alpha- and Gammaherpesvirinae, respectively. Computer comparisons of the amino acid sequences of proteins predicted from the published complete VZV and EBV DNA sequences resulted in the detection of EBV counterparts to 29 of the 67 unique VZV genes. Conserved genes were detected only in the UL component of each genome, and are located in three major regions, within which conserved genes are generally colinear. However, the three regions are arranged differently in the two genomes. These results make it possible in principle to propose the functions of EBV genes on the basis of the functions of their VZV counterparts. The data also allow identification of the types of events which may have occurred during divergence of VZV and EBV, as representatives of the Alpha- and Gammaherpesvirinae, from a common ancestor. PMID- 3033127 TI - Epstein-Barr virus-specific transcription in normal and malignant nasopharyngeal biopsies and in lymphocytes from healthy donors and infectious mononucleosis patients. AB - Cytoplasmic RNA was prepared from biopsy material obtained from the nasopharynx of normal individuals and nasopharyngeal carcinoma (NPC) patients. Similar RNA preparations were prepared from lymphocytes of healthy donors and infectious mononucleosis patients. RNA was radioactively labelled in vitro and hybridized to cloned fragments of B95-8 Epstein-Barr virus (EBV). In all seropositive cases the minimum pattern of EBV-specific RNA expression was like that observed previously in latently infected, EBV-positive lymphoblastoid cell lines or Burkitt's lymphoma biopsies. A novel observation was that all of the control biopsies from normal nasopharynxes expressed EBV-specific RNA, some of which appeared to be associated with a more active state of EBV infection. The pattern of expression in NPC patients was similar to that of the normal donors but showed some variations in complexity. In general the virus-specified small RNAs were not present in normal nasopharyngeal tissue but were expressed in the NPC biopsies. PMID- 3033129 TI - Inhibitory effect of papaverine on HVJ (Sendai virus) replication in rat glioma C6 cells. AB - The replication of HVJ (Sendai virus) in C6 rat glial cells was found to be inhibited by treatment of the cells with papaverine, an inhibitor of cAMP phosphodiesterase, but not with cAMP or dibutyryl cAMP. In addition, cyclic GMP which often manifests a reciprocal relationship to cAMP did not counteract the inhibition of HVJ yield by papaverine. Both viral genome replication and transcription were suppressed slightly by treatment of the cells with papaverine. In the cells cultured in the presence of papaverine, the synthesis of viral proteins and their phosphorylation occurred at normal rates. Membrane immunofluorescence and cell surface immunoprecipitation showed that the viral glycoproteins HN and F0 were expressed on the cell surface of the papaverine treated cells. Moreover, all the viral structural proteins were associated with plasma membrane isolated from the treated cells. These results indicate that papaverine treatment suppresses some part of the process of virus budding at the plasma membrane. PMID- 3033128 TI - Suppression of delayed type hypersensitivity to herpes simplex virus type 1 following immunization with anti-idiotypic antibody: an example of split tolerance. AB - Intraperitoneal (i.p.) immunization with herpes simplex virus type 1 (HSV-1) or an anti-idiotypic antibody (anti-id C) prepared against a monoclonal antibody specific for glycoprotein C of HSV-1, tolerizes mice for an HSV-1 delayed type hypersensitivity (DTH) response. This tolerization could be adoptively transferred to naive X-irradiated mice by splenic T cells, and was specific for HSV-1 DTH. Thus, DTH-tolerized mice responded to vaccinia virus or HSV-2 challenge, while remaining tolerized for HSV-1 DTH. In addition, these animals demonstrated a form of split tolerance such that HSV antibody, cytotoxic T cell and lymphoproliferative responses were detected in vitro. Thus, anti-id C induced a T cell population capable of specifically suppressing the HSV-1 DTH response, mimicking the effect of i.p. and intravenous immunization with HSV-1 found previously. PMID- 3033131 TI - A major phosphoprotein of cells infected with pseudorabies virus is phosphorylated by cellular casein kinase II. AB - Endogenous protein phosphorylation was studied in extracts of hamster fibroblasts infected with pseudorabies virus. The major phosphorylation was detected quite late in infection and involved an acidic protein of Mr 62,000. It was catalysed by an enzyme activity with the properties of cellular casein kinase II. Two dimensional gel analysis was used to demonstrate that this same protein was also phosphorylated in vivo. The phosphoprotein was detected in mature virions and is most likely viral in origin. PMID- 3033132 TI - Generation of a herpes simplex virus type 1 variant devoid of XbaI sites. AB - Using both selection enrichment and site-directed mutagenesis, a herpes simplex virus type 1 (HSV-1) strain 17 genome lacking all four XbaI sites has been generated. The site at 0.45 map units which lies within the gene coding for a polypeptide of 28,000 molecular weight was removed by selection enrichment, while the site at 0.29 map units which lies within the gene coding for glycoprotein H was removed by site-directed mutagenesis. The parental virus from which these two XbaI sites were deleted had previously had the sites at 0.07 and 0.63 map units removed through selection enrichment. The variant devoid of XbaI sites (X4) showed normal growth characteristics; its phenotype was normal apart from the absence of the thymidine kinase protein, which is believed to be unrelated to the loss of XbaI sites. PMID- 3033130 TI - PGJ2, a new antiviral prostaglandin: inhibition of Sendai virus replication and alteration of virus protein synthesis. AB - Prostaglandin J2 (PGJ2) was found to suppress dramatically Sendai virus replication in African green monkey kidney cells in culture. PGJ2 was not toxic at the active dose to uninfected cells and did not significantly inhibit macromolecular synthesis, but it specifically stimulated the synthesis of a polypeptide of 74,000 mol. wt. In Sendai virus-infected cells, PGJ2 partially inhibited virus protein synthesis and caused an alteration in the mobility of the virus glycoprotein HN in SDS-PAGE, corresponding to a decrease of about 4000 in its mol. wt. We propose that the PGJ2-induced alteration in the molecular structure of the HN protein prevents the insertion of this protein into the cell membrane thereby blocking virus maturation. The alpha,beta-unsaturated carbonyl group in the cyclopentane ring of the PGJ2 molecule may be necessary for antiviral activity. PMID- 3033134 TI - Isolation and characterization of a cold-sensitive strain of coxsackievirus A10. AB - A coxsackievirus A10 strain, isolated from a clinical specimen from a patient with pharyngitis, was characterized with respect to its growth properties in different cultured cells and at different incubation temperatures. This virus multiplied within cultured cells and produced cytopathogenic effects, whereas a prototype strain of coxsackievirus A10 did not. The isolate multiplied efficiently in cultured cells at 37 degrees C but its replication was markedly restricted at 32 degrees C. Temperature shift experiments indicated that the cold sensitive event affected the late function(s) of the virus. PMID- 3033133 TI - Transmission of mouse thymic virus. AB - Mouse thymic virus (MTV) is a naturally occurring herpesvirus of mice which produces persistent infection in salivary glands. No transmission study has been reported previously. In the present work, transmissibility of MTV has been studied by close contact between cage-mates, by the transplacental route from experimentally infected pregnant mice to their foetuses at term or delivered by Caesarean section and by nursing mothers to their sucklings. Transmission of MTV was detected between cage-mates after a long period of contact. The virus was also recovered from newborns nursed by infected mothers inoculated 1 day post delivery. However, no transmission was detected in the foetuses following infection of mothers at different stages of pregnancy. PMID- 3033135 TI - Dependence of guanidine sensitivity of poliovirus replication on the concentration of monovalent cations in the culture medium. AB - An earlier suggestion that guanidine may inhibit picornavirus replication by interfering with a monovalent cation-mediated event was tested by determining the effect of varying monovalent cation concentration in isotonic medium on the sensitivity of poliovirus replication in HeLa cells to 0.2 mM-guanidine. Lowering [Na+] in the medium to 50 mM had no effect on virus replication. It was found that the degree of inhibition of virus replication by 0.2 mM-guanidine was inversely related to [Na+] in the medium: 99.8%, 99.1%, 38% and 0% inhibition in the presence of 50, 75, 100 and 145 mM-Na+ respectively. Likewise, guanidine uptake by HeLa cells was also inversely related to [Na+] in the medium. On the other hand, lowering medium [Na+] to 50 or 75 mM resulted in reduced intracellular [Na+] and [K+]. The increased sensitivity of virus replication to guanidine in the presence of low Na+ medium could be abolished with excess K+ in such medium. Excess K+ in low Na+ medium restored intracellular [Na+] and reduced guanidine uptake. Thus, the increased sensitivity of poliovirus replication to guanidine in the presence of low Na+ medium correlated with reduced intracellular [Na+] and [K+] and elevated guanidine uptake. PMID- 3033136 TI - Preliminary studies on antigenic variation of poliovirus using neutralizing monoclonal antibodies. AB - Cross-neutralization assays were done using 85 strains of poliovirus type 1 with five groups of monoclonal antibodies. These strains were classified into 10 subgroups which had marked differences in antigenicity. Subgroups P1-2 (28%) and P1-5 (43%) were dominant and have been epidemic in China in recent years. These two subgroups were antigenically different from the Sabin-1 strain, but according to their responses to one group of monoclonal antibodies they had antigenic epitopes in common with the Mahoney and Brunhilde strains. Similarly, 91 strains of type 3 poliovirus were classified into six subgroups with another five groups of monoclonal antibodies. The results showed that strain P3/Yunnan/2/84, which was isolated from cases of poliomyelitis in a local epidemic in the Yunnan province of China in 1984, and strain P3/Finland/23127/84, which was isolated in Finland in 1984, were both antigenically different from the Sabin-3 strain and the reference virulent strain. PMID- 3033137 TI - Intracellular RNAs of the feline infectious peritonitis coronavirus strain 79 1146. AB - In Felis catus whole foetus D cells infected with feline infectious peritonitis virus (FIPV), strain 79-1146, six virus-specific, poly(A)-containing RNA species of about 20, 9.6, 5.2, 3.8, 2.8 and 1.6 kb were found. By translation in vitro the 3.8 and 2.8 kb RNAs were shown to encode the 25K envelope protein and the 45K nucleocapsid protein, respectively. The partial map of the FIPV genome was compared with genomic maps of porcine, murine and avian coronaviruses. Differences in these maps suggest that transcription units have been lost or gained during coronavirus divergence. PMID- 3033138 TI - The two major structural phosphoproteins (pp65 and pp150) of human cytomegalovirus and their antigenic properties. AB - Human cytomegalovirus (HCMV) purified from cell culture contains two dominant structural phosphoproteins with apparent molecular weights of 65,000 and 150,000, designated as pp65 and pp150 respectively. The humoral immune response of infected individuals against pp65 is relatively weak and is not always detectable by Western blot analyses. This report shows that recent clinical isolates of HCMV do not necessarily have pp65 as a prominent constituent, suggesting that the low immune reaction is due to variable expression of the pp65 in natural infections. However, the HCMV strains tested in this study produced the large structural phosphoprotein (pp150) in about equal amounts. The pp150 is remarkably immunogenic, if compared with all other virion constituents; serum pools and individual sera from HCMV-infected patients recognized this particular protein intensively in immunoblot assays. Thus, phosphoprotein pp150 seems to be the primary polypeptide candidate for expression cloning in order to develop reagents for novel ways of HCMV diagnosis. PMID- 3033139 TI - A herpes simplex virus type 1 variant which fails to synthesize immediate early polypeptide VmwIE63. AB - We report the isolation of a variant (X2D) of herpes simplex virus type 1 strain 17 which has a deletion of 5 X 10(6) mol. wt. in the long unique and long inverted repeat regions, such that one copy of the immediate early (IE) gene 1 and two unique open reading frames coding for polypeptides of 20K and 22K are deleted. The mutant X2D synthesizes reduced levels of VmwIE110, and also apparently fails to synthesize VmwIE63, at both the protein and RNA levels, despite there being no apparent deletion in the coding or controlling regions of the IE2 gene. X2D also fails to synthesize the thymidine kinase polypeptide but exhibits normal growth characteristics in tissue culture. PMID- 3033140 TI - Identification of human papillomavirus type 18 E6 polypeptide in cells derived from human cervical carcinomas. AB - We recently reported the expression of human papillomavirus type 18 (HPV-18) E6 protein in bacteria and the production of anti-E6 polyclonal antibodies. This work has now been extended with the production of a panel of monoclonal antibodies against the HPV-18 E6 protein. These antibodies demonstrate that there is little antigenic conservation in the E6 protein between HPV-16 and HPV-18, with only one antibody recognizing a cross-reactive epitope. We have used both the monoclonal and the polyclonal antibodies to look for E6 expression in a number of HPV DNA-containing cell lines. These reagents specifically detected a 16.5K mol. wt. polypeptide in cells derived from a human cervical carcinoma. PMID- 3033141 TI - Analysis of Marek's disease virus serotype 1-specific phosphorylated polypeptides in virus-infected cells and Marek's disease lymphoblastoid cells. AB - By use of monoclonal antibodies, a virus-specific cytoplasmic antigen related to phosphorylated polypeptides specific to serotype 1 of Marek's disease virus (MDV) related viruses (MDV1) has been identified in all MD tumour cell lines examined, as well as in infected cells and in tumour lesions of chickens with MD. At least two phosphorylated polypeptides with mol. wt. 39,000 (39K) to 36K and 24K (pp39/36 and pp24, respectively) were identified in the MD tumour cell line H10 cultured at 33 degrees C by immunoprecipitation with monoclonal antibody M21 which reacts with virus-specific phosphorylated polypeptides. These polypeptides were not detected in cells infected with MDV-related viruses of serotype 2 or 3. Immunoblot analysis indicated that these two polypeptides contained a serotype 1 specific epitope recognized with M21. An additional 41K polypeptide appeared in different virus strains of serotype 1. These polypeptides were found to contain phosphorylated serine but no detectable phosphorylated tyrosine or phosphorylated threonine. Cell fractionation indicated that the two phosphorylated polypeptides were mainly associated with smooth and rough endoplasmic reticulum fractions of cells infected with MDV1. Furthermore, the mRNA coding for pp39/36 could be separated from that coding for pp24 on a sucrose density gradient. These results suggest that pp24 and pp39/36 are translated from distinct mRNAs and encoded from overlapping genes or separate regions with partial DNA homology in the MDV1 genome. PMID- 3033142 TI - The herpes simplex virus type 1 DNA polymerase gene: site of phosphonoacetic acid resistance mutation in strain Angelotti is highly conserved. AB - By comparative sequence analysis of the herpes simplex virus type 1 DNA polymerase gene of strain Angelotti and a phosphonoacetic acid-resistant (PAAr) derivative, the site of the PAAr mutation was identified as a single nucleotide (C----T) conversion within the mapping limits of the known PAAr mutations of strains KOS and 17. The conservative amino acid change at residue 719 from alanine to valine results in a radical change in the properties of the polymerase, rendering the mutant enzyme resistant to PAA and various antiviral compounds. Amino acid homologies as well as secondary structure analysis reveal that the PAAr mutation is contained in a 14 amino acid sequence which is highly conserved, and detected in the central domain of prokaryotic and eukaryotic DNA polymerases. PMID- 3033143 TI - Virulence is not conserved in recombinants between herpes simplex virus types 1 and 2. AB - The virulence of 31 herpes simplex virus type 1 X herpes simplex virus type 2 intertypic recombinants was determined following intraperitoneal inoculation into CBA mice. Only eight of the recombinants killed any of the mice and of these, only one recombinant was as virulent as its type 2 parent, the other seven recombinants being intermediate between their type 1 and type 2 parental viruses in virulence. These results indicate that most heterotypic combinations are attenuated independently of the virulence of the parental viruses and therefore that the virulence of HSV is controlled multigenically. PMID- 3033144 TI - Varicella-zoster virus specifies a thymidylate synthetase. AB - A homology search of proteins predicted from the recently reported complete DNA sequence of varicella-zoster virus (VZV) revealed that the product of gene 13 was highly homologous to eukaryotic and prokaryotic thymidylate synthetases (TSs). The VZV protein was shown to be a TS by three functional tests. Firstly, a plasmid designed to express the native protein was able to complement a strain of Escherichia coli in which the natural TS gene is deleted. Secondly, in an enzyme assay for TS, extracts of the complemented strain were capable of releasing tritiated water from 2'-deoxy[5-3H]uridylate. Thirdly, these extracts contained a protein that bound isotopically labelled 5-fluoro-2'-deoxyuridylate, a ligand specific for the active site of TS. In addition, a novel ligand-binding protein was detected in human cells infected with VZV. PMID- 3033145 TI - Extension and retraction of axonal projections by some developing neurons in the leech depends upon the existence of neighboring homologues. I. The HA cells. AB - The role of homologues in the establishment of the pattern of axonal projections of identified segmentally homologous neurons was investigated by means of selective cell ablation and dye injection. The cells studied were the bilateral pairs of heart accessory (HA) neurons found in the fifth and sixth segmental ganglia of the leech ventral nerve cord. Homologues start their morphological differentiation with identical axonal projections, and segmental differences are manifested later, when specific branches stop growing and disappear. The deletion of single HA cells at early stages, however, permits these branches to survive in their ipsilateral homologues and to grow and take over the projections of the deleted neurons. In addition, if both HA homologues on the same side of the nerve cord, or three of the four HA cells, are deleted in an animal, the remaining HA cells often extend novel projections. These observations suggest that either competition for targets, inputs or growth factors, or direct interactions among homologous cells may play a role in the differentiation of segment specific patterns of axonal projections. PMID- 3033146 TI - Characterization of the binding of [3H]SR 95531, a GABAA antagonist, to rat brain membranes. AB - A synthetic derivative of gamma-aminobutyric acid (GABA), SR 95531 [2-(3'-carboxy 2'-propyl)-3-amino-6-p-methoxyphenylpyridazinium bromide], has recently been reported, on the basis of biochemical and in vivo microiontophoretic studies, to be a potent, selective, competitive, and reversible GABAA antagonist. In the present study, the binding of [3H]SR 95531 to washed, frozen, and thawed rat brain membranes was characterized. Specific binding was linear with tissue concentrations, had a pH optimum at neutrality, and was maximal at 4 degrees C after 30 min of incubation. Pretreatment of the membranes with Triton X-100 resulted in a 50% decrease of specific binding. Addition of iodide, thiocyanate, or nitrate to the incubation mixture decreased the affinity of [3H]SR 95531 for its binding site; Na+ had no effect. Subcellular fractionation showed that 74% of the P2 binding was in synaptosomes; 31% of the total homogenate binding was in P2 and 50% in P3. The binding of [3H]SR 95531 was saturable; Scatchard analysis of the saturation isotherm revealed two apparent populations of binding sites (KD of 6.34 nM and Bmax of 0.19 pmol/mg of protein; KD of 32 nM and Bmax of 0.81 pmol/mg of protein). The binding of [3H]SR 95531 was reversible, and association and dissociation kinetics confirmed the existence of two binding sites. Only GABAA ligands were effective displacers of [3H]SR 95531. GABAA antagonists were relatively more potent in displacing [3H]SR 95531 than [3H]GABA; the inverse was true for GABAA agonists. There were marked regional differences in the distribution of binding sites: hippocampus = cerebral cortex greater than thalamus = olfactory bulb = hypothalamus = amygdala = striatum greater than pons medulla and cerebellum. The surprisingly low density of binding sites in the cerebellum was owing to a marked reduction of Bmax values at both the high- and the low-affinity binding sites. In conclusion, the present results demonstrate specific, high-affinity, saturable, and reversible binding of [3H]SR 95531 to rat brain membranes and strongly suggest that this radioligand labels the GABAA receptor site in its antagonist conformation. PMID- 3033147 TI - Influence of hydrocortisone on chick embryo retina development. AB - Treatment of chick embryos in ovo with hydrocortisone-21-phosphate (a single dose of 150 micrograms) caused a marked reduction of retinal thymidine kinase activity 24 h later. The inhibitory effect was highest (65-70%) in 8-10-day-old embryos and declined with age, disappearing after day 15. It was accompanied by a reduction in thickness of the retinal layers. Adrenocorticotropic hormone (ACTH) treatment (10 micrograms daily for 2 days) also produced an age-dependent inhibitory effect on retinal thymidine kinase, whereas treatment with a single dose of 200 micrograms of metopirone, a compound that prevents the 11 beta hydroxylation of steroid molecules in the adrenal glands, impeded the decrease in thymidine kinase activity that normally occurs in chick embryo retina after day 9 of development. In addition, metopirone prevented the inhibition exerted by ACTH on thymidine kinase activity but had no effect on the action of hydrocortisone. PMID- 3033148 TI - Independent protein kinases associated with the rat cerebral synaptic junction: comparison with cyclic AMP-dependent and Ca2+/calmodulin-dependent protein kinases in the synaptic junction. AB - Independent protein kinases in the synaptic junction (SJ) isolated from rat cerebrum were characterized. SJ showed a protein kinase activity, phosphorylating intrinsic proteins, even in the absence of cyclic AMP or Ca2+ plus calmodulin (CaM) exogenously added. The activity was affected neither by Ca2+ concentrations in the physiological fluctuation range nor by the addition of specific ligands such as glutamate, aspartate, acetylcholine, and concanavalin A. The activity was not due to cyclic AMP-dependent protein kinase in SJ, since the activity was not inhibited by an inhibitor protein for cyclic AMP-dependent protein kinase, and since synapsin I was not specifically phosphorylated whereas cyclic AMP-dependent kinase appeared to phosphorylate selectively the protein in SJ. Phosphorylation of SJ proteins by the independent kinases was about one-third of that of the Ca2+/CaM-dependent protein kinase intrinsic to SJ. The apparent Km for ATP was estimated to be 700 microM. Proteins of 16K Mr and 117K Mr were specifically phosphorylated under the basic condition (in the absence of the substances known to activate specifically protein kinases), as well as six other proteins both under the basic conditions and in the presence of Ca2+ and CaM. The phosphorylation of 150K Mr, 60K Mr, 51K Mr, and 16K Mr SJ proteins was enhanced after prephosphorylation of SJ proteins by intrinsic kinase in the presence of Ca2+ and CaM.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3033149 TI - Oligodendroglial differentiation in glial primary cultures: requirement for mevalonate. AB - The oligodendroglial enzyme, 2',3'-cyclic nucleotide 3'-phosphohydrolase (CNP), is a valuable marker for expression of oligodendroglial differentiation in glial primary cultures, and the inducibility of this enzyme by dibutyryl-3',5'-cyclic AMP (dBcAMP) appears to be limited to immature or developing oligodendroglia. To investigate the relationship between the induction of CNP and the sterol biosynthetic pathway, primary cultures of glia dissociated from the brains of newborn rats were maintained in 10% fetal calf serum (FCS) and exposed to 1 mM dBcAMP on day 7 in culture. Cultures so treated for either 48 h or 72 h demonstrated a three- to fourfold induction of CNP specific activity. The magnitude of this induction was not affected when the cholesterol content of the culture medium was reduced by greater than 95% by placing the cultures in 10% lipoprotein-poor serum rather than 10% FCS during the exposure to dBcAMP. Mevinolin (10 microM), a specific inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase, the rate-limiting enzyme of the sterol biosynthetic pathway, completely inhibited the induction of CNP by dBcAMP, while not affecting either the accumulation of cellular protein per flask or rate of protein synthesis. Simultaneous addition of mevalonate (20 mM) prevented the inhibition of the induction of CNP by mevinolin. However, simultaneous addition of low density lipoprotein sufficient to increase the cholesterol content of the medium 80-fold failed to correct mevinolin's inhibition of the induction of CNP.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3033150 TI - Alterations in lipid metabolism, Na+,K+-ATPase activity, and tissue water content of spinal cord following experimental traumatic injury. AB - Traumatic spinal cord injury has recently been shown to cause a rapid increase in free fatty acids (FFAs) and lipid degradation in cats. The present studies report a more delayed, time-dependent increase in FFAs and a concomitant decrease in phospholipids following traumatic spinal injury in rats. The largest percentage increases were found for polyunsaturated fatty acids, particularly arachidonic acid. Associated with these changes were a reduction in the activity of Na+,K+ ATPase and development of spinal cord edema. These findings support the hypothesis that traumatic spinal cord injury leads to delayed, as well as early, hydrolysis of membrane phospholipids, resulting in the liberation of FFAs. Such changes may contribute to secondary spinal cord injury either through direct effects on membranes or through the actions of secondary metabolic products such as the eicosanoids. The latter may cause tissue injury by contributing to the reduction in spinal cord blood flow or through inflammatory responses that follow trauma. PMID- 3033152 TI - Different behaviors among lysosomal enzymes in the cerebellum of jaundiced Gunn rats with cerebellar hypoplasia. AB - Activities of six lysosomal enzymes in the cerebellum of jaundiced homozygous (jj) Gunn rats were examined from 5 to 20 days of life and compared with those in heterozygotes (j+). Significantly higher enzyme activities were first detected at 8 days. The jj/j+ activity ratios of all enzymes peaked at 15 days. The ratios of beta-glycerophosphatase, beta-mannosidase, and acid lipase were only 1.3-1.7, whereas those of arylsulfatase and cathepsin were 2.0 and 3.1, respectively. The most striking increase in activity was observed with beta-glucuronidase, the ratio of which was 8.4. These results indicate a selective increase in activities of certain lysosomal enzymes in the hypoplastic cerebellum of jj rats. PMID- 3033151 TI - Effects of chronic ethanol treatment on the beta-adrenergic receptor-coupled adenylate cyclase system of mouse cerebral cortex. AB - Chronic ingestion of ethanol, which produced tolerance and physical dependence, resulted in altered function of the cerebral cortical beta-adrenergic receptor coupled adenylate cyclase system in mice. Although there was no change in basal adenylate cyclase activity, or in the activity of the digitonin-solubilized catalytic unit, stimulation of adenylate cyclase activity by the nonhydrolyzable guanine nucleotide analog guanylylimidodiphosphate [Gpp(NH)p] was reduced in brains of ethanol-fed animals. Ethanol added in vitro increased adenylate cyclase activity, and this enhancement, in the presence of Gpp(NH)p, was also reduced in cortical membranes of ethanol-fed mice. Furthermore, the maximal response to isoproterenol was decreased, and the EC50 for isoproterenol stimulation of adenylate cyclase activity was increased in ethanol-fed animals. The results are consistent with a qualitative or quantitative defect in the function of the stimulatory guanine nucleotide-binding protein (Ns), as well as in the beta adrenergic receptor, after chronic ethanol exposure. In part, these changes appear to be similar to those that occur during heterologous desensitization of various receptor systems, and may be associated with dependence on or tolerance to ethanol. PMID- 3033153 TI - Subcellular distribution of ependymins in goldfish brain measured by radioimmunoassay. AB - Goldfish CNS was fractionated by differential and density gradient centrifugation. The fractions obtained were characterized by marker enzymes typical of various subcellular organelles. They were further analyzed by radioimmunoassay for their contents of ependymins, two CNS glycoproteins known to participate in biochemical reactions after learning events. Ependymins were shown to be major constituents of the soluble cytoplasm (5.6% of the total protein content). The nuclear fraction was virtually devoid of ependymins (0.6% of protein). Small amounts were observed in the crude synaptosomal and microsomal fractions (1.0 and 3.5%, respectively). The highest steady-state concentration of ependymins, however, was measured in the brain extracellular fluid (15.6% of the protein), including the CSF. The specificity of the distribution was examined by intracerebroventricular injection of 125I-labeled ependymins as exogenous marker substances. No indication of an artificial redistribution of the radiolabel during homogenization and fractionation was obtained. The exogenous analogues of ependymins were, however, incorporated in vivo into organelles recovered in the nuclear and crude synaptosomal fractions. Our results suggest that ependymins may interact with synaptic membranes from the extracellular fluid, although so far no evidence for a specific receptor-type binding site could be obtained in vitro. PMID- 3033155 TI - Enhanced coupling of neonatal muscarinic receptors in rat brain to phosphoinositide turnover. AB - The relationship between the density of the muscarinic receptor in developing rat cerebral cortex and its coupling to phosphoinositide turnover is examined. Tissue slices from rats of various ages were incubated with myo-[2-3H]inositol, and the effect of carbamoylcholine on the release of total inositol phosphates was determined. Binding of [3H]quinuclidinyl benzilate was determined in the same tissue. Although muscarinic receptor density in day-18 embryonic cortex was only 5% of that in the adult, the maximal response of stimulated phosphoinositide turnover to carbamoylcholine (1-10 mM) was at the adult level (i.e., three-fold increase). Comparison of the dependence of the turnover on carbamoylcholine concentration revealed that in neonates, the dose-response curve was shifted to the left, giving a half-maximal effect at concentrations approximately tenfold lower than that in the adult. In addition, the partial muscarinic agonists oxotremorine-2 and bethanechol were both more efficacious in young rats than in adults. The differences could not be accounted for either by alterations in agonist affinity for the receptor or by the presence of "spare" muscarinic receptors. These results indicate that muscarinic receptors in fetal and newborn rat cerebral cortex are more efficiently coupled to stimulation of phosphoinositide turnover than in the adult. PMID- 3033154 TI - Glycoprotein as a constituent of purified gamma-aminobutyric acid/benzodiazepine receptor complex: structures and physiological roles of its carbohydrate chain. AB - The effect of treatments with various enzymes and chemically modifying agents on [3H]muscimol binding to a purified gamma-aminobutyric acid (GABA)/benzodiazepine receptor complex from the bovine cerebral cortex was examined. Treatments with pronase, trypsin, guanidine hydrochloride, and urea significantly decreased the binding of [3H]muscimol, but dithiothreitol, N-ethylmaleimide, reduced glutathione, oxidized glutathione, cysteine, and cystine had no significant effect. These results indicate that the GABA receptor indeed consists of protein, but -SH and -S-S- groups in the protein are not involved in the exhibition of the binding activity. On the other hand, column chromatography using concanavalin A Sepharose eluted protein having [3H]muscimol binding activity and staining of glycoprotein using an electrophoresed slab gel indicated the existence of two bands originating from the subunits of the GABA/benzodiazepine receptor complex. Furthermore, treatments with various glycosidases such as glycopeptidase A, beta galactosidase, and alpha-mannosidase significantly increased the binding of [3H]muscimol. These results strongly suggest that GABA/benzodiazepine receptor complex is a glycoprotein and that its carbohydrate chain may be a hybrid type. Treatment with beta-galactosidase resulted in the disappearance of the low affinity site for [3H]muscimol binding and in an increase of Bmax of the high affinity site, without changing the KD value. These results suggest that the carbohydrate chain in the receptor complex may have a role in exhibiting the low affinity binding site for GABA. The observation that the enhancement of [3H]muscimol binding by treatments with beta-galactosidase and glycopeptidase A were much higher than that with alpha-mannosidase may also indicate a special importance of the beta-galactosyl residue in the inhibition of GABA receptor binding activity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3033156 TI - Distant metastases from a malignant glioma: unusual complications associated with treatment of a glioblastoma: distant metastases and focal white matter degeneration. PMID- 3033157 TI - Cyclosporine enhances virally induced T-cell-mediated demyelination. The effect of cyclosporine on a demyelinating virus infection. AB - Semliki Forest virus infection of adult mice results in a demyelinating meningoencephalomyelitis. Demyelination does not result from direct viral damage but from the activity of T lymphocytes. We have studied the effect of the immunosuppressant cyclosporine (Cs) on the outcome of this infection. Cs had no effect when given 5 days after the infection, and little effect when given 4 h after infection. When the Cs was given 48 h before infection there was a prolongation of the blood and brain virus titres, and a reduction in some mice of serum IgG anti-viral antibody synthesis, but an increase in the severity of the CNS inflammatory response and the demyelination. Consideration of these findings along with measurement of Cs levels in the serum and cerebrospinal fluid suggests that this drug does not cross the blood brain barrier. PMID- 3033158 TI - Relation of renin-angiotensin system activity to left ventricular hypertrophy and function in experimental and human hypertension. AB - This article examines available data concerning the hypothesis that the renin angiotensin system directly stimulates cardiac hypertrophy and dysfunction in hypertension. Several experiments support a direct effect of angiotensin on myocardial protein synthesis and suggest that this may be independent of adrenergic influences. However, the role played by the renin system in the pathogenesis of hypertensive cardiac hypertrophy may be small, for left ventricular (LV) muscle mass is not systematically greater in high-renin as opposed to low-renin forms of experimental or human hypertension. In animal studies, regression of LV hypertrophy is more consistently observed in response to drugs that inhibit as opposed to those that stimulate renin-angiotensin system activity (i.e., converting enzyme inhibitors or beta-adrenoceptor blockers vs. diuretics or direct vasodilators), although the difference is more quantitative than absolute. Similarly, significant reductions in LV mass occurred in 13 of 16 human trials with converting enzyme inhibitors or beta blockers as opposed to 2 of 10 trials with diuretics or vasodilators (mean = 7.11; p less than 0.01). However, uncertainties regarding the degree of reduction in angiotensin II and adrenergic effects, possible interactions between these systems, and incomplete characterization of induced changes in hemodynamic load on the heart all limit the interpretation of available studies. Preliminary data suggest that an inverse relation exists between renin-angiotensin system activity and LV systolic performance in primary and secondary human hypertension, a possibility that merits further study. PMID- 3033159 TI - Operation for small-cell carcinoma revisited. PMID- 3033161 TI - High-dose induction chemotherapy with cyclophosphamide, etoposide, and cisplatin for extensive-stage small-cell lung cancer. AB - To exploit possible dose-response and combination drug synergism, 20 previously untreated patients with extensive-stage small-cell lung cancer (SCLC) received one or two courses of high-dose induction chemotherapy consisting of cyclophosphamide (100 mg/kg), etoposide (1,200 mg/m2), and cisplatin (120 mg/m2) (HDCEP). HDCEP was followed by four cycles of standard-dose cyclophosphamide (1,000 mg/m2), doxorubicin (40 mg/m2), and vincristine (1.4 mg/m2) (CAV). Response was determined after HDCEP and following CAV. Reevaluation included repeat bronchoscopy and chest computerized tomography (CT), as well as repetition of all initially abnormal studies. All patients were evaluable for response and toxicity. Overall response to HDCEP was 90%, with a complete response (CR) rate of 65% (95% confidence limits, 44% to 86%) and a partial response (PR) rate of 25% (95% confidence limits, 6% to 44%). All patients either maintained or improved their initial response while receiving CAV. Median duration of response was 6 months (range, 2 to 12 months) and median survival was 9.5 + months (range, 2 to 21 + months). All 37 courses of HDCEP were associated with leukopenia (less than 1,000/microL), 92% with thrombocytopenia (less than 20,000/microL), and 84% with fever of greater than 38.5 degrees C. Additional toxicities included bacteremia (24%), nausea and emesis (59%), mucositis (57%), diarrhea (38%), and hemorrhagic cystitis (5%). There were two treatment-related deaths due to infection. A third patient died 4 months after completing HDCEP with pulmonary fibrosis. Although response duration and median survival were not improved, HDCEP produced a high CR rate in ambulatory patients with extensive-stage SCLC. PMID- 3033160 TI - The role of surgery in the management of selected patients with small-cell carcinoma of the lung. AB - This study was designed to evaluate the efficacy of surgical resection of the primary tumor and lymph nodes in patients with localized small-cell carcinoma who had responded to induction chemotherapy. The study was performed in 37 patients who received two cycles of chemotherapy consisting of cyclophosphamide, doxorubicin, and etoposide. Those patients who achieved a complete or partial (greater than 50%) response were evaluated for thoracotomy and the primary tumor and regional lymph nodes excised when feasible. Postoperatively, the patients received prophylactic cranial irradiation and were maintained on the same chemotherapy for an average of 11 months. Twelve patients were resected and found to have residual small-cell carcinoma in the operative specimen (ten) or no residual disease (two). Seven of these patients (58%) are alive without evidence of disease (median follow-up, 24 months). Seven other patients who were resected proved to have either residual foci or small-cell carcinoma mixed with adenocarcinoma or large-cell carcinoma (four) or only focal areas of adenocarcinoma, large-cell carcinoma, or squamous-cell carcinoma with no evidence of residual small-cell carcinoma. Five of these patients (71%) are alive without evidence of disease (median follow-up, 36 months). Two of the 16 patients who were not resected but treated with chemotherapy and radiation are alive at 15 and 31 months without evidence of disease, the other 14 are dead of disease. PMID- 3033162 TI - Cytologically negative pericardial effusion complicating combined modality therapy for localized small-cell carcinoma of the lung. AB - A 17% frequency of cytologically negative pericardial effusion (CNPE), accompanied in some cases by tamponade, occurred a median of 12.6 months from the onset of treatment for localized small-cell carcinoma of the lung. CNPE was apparently caused by toxicity of radiation/chemotherapy treatment rather than recurrent cancer. The occurrence of CNPE does not appear to represent enhanced toxicity of immediate (as opposed to delayed) concurrent chemoradiotherapy, but may be a consequence of the superior survival status of patients treated in this way. The onset of chest pain and/or dyspnea associated with increase in cardiac silhouette and positive echocardiogram allowed accurate diagnosis. Each instance was relatively easily managed by catheter drainage, and, for some patients, with the addition of nonsteroidal antiinflammatory drugs. It is important to recognize the possibility that a radiation/chemotherapy-related syndrome of pericardial effusion/tamponade may occur so that early diagnosis can be made and the risk of fatal tamponade avoided. PMID- 3033163 TI - Brain tumor resection guided by intraoperative computed tomography. AB - Intraoperative computed tomography (CT) with a dedicated, therapeutic scanner facilitated resection of intracranial tumors in nine patients. Major benefits of intraoperative CT included exact, multiplanar lesion localization, maximal surgical resection and immediate recognition of potential intraoperative complications such as intracranial hemorrhage. Three cases are presented in detail to demonstrate the surgical technique. No complications related to the use of intraoperative imaging occurred. Although total resection of malignant brain neoplasms remained an unrealized goal, intraoperative CT increased the surgical accessibility of malignant and benign lesions located in critical areas of the brain. PMID- 3033165 TI - The probabilistic nature of synaptic transmission at a mammalian excitatory central synapse. AB - The synaptic connection between single group I afferents and dorsal spinocerebellar tract (DSCT) neurons in the cat spinal cord has been studied in an attempt to gain insight into the mechanisms of excitatory synaptic transmission in the mammalian CNS. Fluctuations in the amplitude of single group I fiber EPSPs in DSCT neurons were examined using a numerical deconvolution procedure to reduce the effects of contaminating noise. In general, it was found that single fiber EPSPs fluctuate in peak amplitude between discrete levels separated by equal or quantal increments. Many previous studies have proposed simple binomial statistics as a general model of quantal synaptic transmission. In the present study we show that simple binomial statistics do not describe the fluctuations in amplitude of single group I fiber EPSPs in DSCT neurons. It is suggested that nonuniformities in the probability of transmitter release from release site to release site explain the failure of the binomial model to describe the EPSP fluctuation pattern at this synapse. Nonuniform quantal transmission is proposed as a more adequate description of excitatory synaptic transmission in the mammalian CNS. PMID- 3033164 TI - Misonidazole and CCNU chemotherapy for recurrent primary brain tumor. AB - CCNU chemotherapy prolongs survival of patients with primary brain tumor when given at the time of tumor progression following radiation therapy. Used as single agent, response rates of 30 to 80 per cent have been reported with median response durations of five to six months. Experimentally, tumor cytotoxicity is enhanced using the combination of misonidazole and CCNU, without increasing myelotoxicity. In this phase I/II study, 23 patients with primary brain tumor which progressed following radiation therapy were treated with combined CCNU and misonidazole. In all patients either the diagnosis of high grade glioma was made at the time of initial diagnosis prior to radiation therapy or the tumor transformed from low grade to high grade glioma at the time of progression following radiation therapy. CCNU 120 mg/M2 was given four hours following misonidazole 3.5 g/M2 every six weeks, with dosage adjustments for myelotoxicity. Treatment was continued for one year or until tumor progression. Of the 17 patients in the study for one year or more, 11 (65 per cent) survived for one year, and six (35 per cent) remained free of tumor progression for one year. Median time to tumor progression from start of CCNU plus misonidazole chemotherapy was 27 weeks and median survival was 80 weeks. No severe complications resulted from myelotoxicity. One patient developed mild peripheral neuropathy which disappeared following discontinuation of misonidazole. PMID- 3033166 TI - Long-term modifiability of anomalous and delayed rectification in guinea pig inferior olivary neurons. AB - Delayed and anomalous rectification was studied in inferior olivary (I.O.) neurons in guinea pig brain stem slices maintained in vitro. Hyperpolarization of the I.O. cell beyond rest membrane potential was accompanied by anomalous rectification (AR). This consisted of 2 parts: an instantaneous and a time dependent component. The "instantaneous" component was blocked by bath addition of Ba2+ or Cs+ and demonstrated inactivation following prolonged hyperpolarization. The time-dependent component, referred to as the gK(ol), was blocked by harmaline in concentrations of 0.1 mg/ml or by substitution of Co2+, Cd2+, or Mn2+ for Ca2+ in the bath. The gK(ol) was blocked by extracellular Cs+ but not by Ba2+. Delayed rectification (DR), consisting of 2 distinct components, was observed after membrane depolarization by more than 10 mV with respect to rest (usually at -65 mV). One of the components of the DR was found to be quite similar to the classical gK. It did not demonstrate significant inactivation with membrane potential change and was reduced by Ba2+ or tetraethylammonium (TEA). A second component of the DR demonstrated voltage-dependent inactivation and was thus referred to as gK(inact). This inactivation determined by current-clamp measurements had a sigmoidal time course, with approximately a 1 sec onset latency and a half-time to peak of 7 sec. The inactivation of gK(inact) outlasted current injection for tens of seconds to several minutes, depending on the duration and amplitude of the preceding depolarization. During this period, I.O. neurons could be easily activated and demonstrated full dendritic spikes following current injection or excitatory synaptic input that had previously been subthreshold for spike initiation. The inactivation component of the DR was removed by prolonged membrane hyperpolarization beyond rest. gK(inact) was blocked by 4-aminopyridine (4-AP; 100 microM) but not by Ba2+. This inactivation was dependent on the presence of extracellular Ca2+ or Ba2+. Addition of Co2+ or Cd2+ to the bath did not block gK(inact) but did prevent its inactivation. The modulatory effects of these different membrane conductances on the integrative properties of I.O. neurons are described. The long duration of the inactivation of DR and AR is considered as the basis for a dynamic long-term modulation of the electroresponsive and integrated properties of I.O. neurons. PMID- 3033167 TI - Mapping of retinal and geniculate neurons onto striate cortex of macaque. AB - A unity ratio between geniculate and ganglion cells can be shown in the macaque visual system. Comparison of the densities (cells/deg2) in the dorsal lateral geniculate nucleus (dLGN) of parvocellular (P) and magnocellular (M) cells, respectively, representing color-opponent and broad-band ganglion cells, with cortical magnification (mm2/deg2) gives the number of afferents per square millimeter in striate cortex (V1). For P cells, this afferent density rises only slightly with eccentricity, indicating that V1 magnification is approximately proportional to the density of P cells. The density of cytochrome oxidase puffs in V1 also rises only slightly with eccentricity. As a result, the number of P cell afferents per puff-centered module is remarkably constant throughout V1. Our findings thus support a novel hypothesis of peripheral scaling, in which V1 cortical magnification is based on the mapping of just 1 class of afferent onto V1 modules. This "P-cell module" in V1 may be composed of submodules corresponding anatomically to the honeycomb cell in layer 4A of V1 and physiologically to a minimal complete set of color-opponent ganglion cells. In contrast, the afferent density of M cells rises steeply with eccentricity, so that the reciprocal of their afferent density, the cortical "domain" of M cells, declines with eccentricity. This decline is similar to that of point-image area in V1. As a result, the number of M cells per point-image area is nearly constant. This quantity is analogous to the receptive-field coverage factor in the retina, which for M cells is fairly constant and greater than unity at all eccentricities. The results show fundamental differences between the neural maps of these 2 major cell types, differences that are likely to have psychophysical consequences. PMID- 3033168 TI - Synapsin I in PC12 cells. II. Evidence for regulation by NGF of phosphorylation at a novel site. AB - NGF treatment of PC12 cells caused a rapid increase in the state of phosphorylation of synapsin I. This phosphorylation of synapsin I is accompanied by a decrease in its electrophoretic mobility on SDS-PAGE. Phosphopeptide fingerprint analysis of the synapsin I revealed that this phosphorylation occurred on a particular phosphopeptide, designated peptide N. Phosphoserine was the only phosphoamino acid detected in peptide N. Partially purified PC12 synapsin I was a substrate for several protein kinases known to be capable of phosphorylating brain synapsin I, but none of these kinases phosphorylated synapsin I on peptide N. The results suggest that the NGF-stimulated phosphorylation of synapsin I may be mediated by a novel protein kinase. PMID- 3033169 TI - Effects of early unilateral blur on the macaque's visual system. II. Anatomical observations. AB - We studied the effects of early unilateral blur on the anatomical organization of the visual pathways in 8 macaque monkeys. Blur was induced in one eye, beginning 2-14 d after birth, by 0.5% atropine twice a day. Atropinization was stopped at 6 8 months of age, and the animals were studied for anatomy 3-24 months later. The retina and all other eye tissues showed normal histology. In the dorsal lateral geniculate nucleus (LGN), cells in parvocellular layers receiving input from the atropine-treated eye were 9-32% smaller and were more lightly stained than those in layers innervated by the untreated eye. These changes were generally larger in the LGN ipsilateral to the treated eye. LGN cell size changes were absent or much smaller in the magnocellular layers. In the striate cortex, the distribution of the oxidative enzyme cytochrome oxidase (CO) was markedly altered in layer 4C beta. Layer 4C beta is uniformly stained in normal animals, but showed a distinct pattern of alternating high and low CO bands in the atropine-treated animals; the bands of higher CO activity were narrower than the bands of lower activity and had a 857-1050 micron repeat. Fainter banding was seen in layers 4A, 4C alpha, and 6, but the density of the rows of dark CO-stained dots in layer 3 was unaffected. Double-labeling revealed that the narrow dark CO bands were associated with the centers of the ocular dominance columns devoted to the atropine-treated eye. The distribution of 14C-2-deoxyglucose uptake in visual cortex produced by 4.5-9 c/deg spatial frequency stimulation was strongly biased toward the untreated eye. The treated eye could, however, elicit reasonably strong uptake when stimulated with patterns containing lower spatial frequencies. These results suggest that unilateral neonatal blur preferentially affects the parvocellular layers of the LGN and layer 4C beta of striate cortex, which are the portions of the central visual system associated with the processing of information concerning fine spatial detail. These anatomical changes are consistent with the high spatial frequency loss of vision demonstrated behaviorally and electrophysiologically in the atropine eye-driven visual system of these same animals. PMID- 3033170 TI - Numbers and proportions of GABA-immunoreactive neurons in different areas of monkey cerebral cortex. AB - The number and proportion of neurons displaying GABA immunoreactivity were determined for 50-micron-wide columns through the thickness of 10 areas of monkey cerebral cortex, including the precentral motor area (area 4), 3 cytoarchitectonic fields of the first somatic sensory area (areas 3b, 1, and 2), 2 areas of parietal association cortex (areas 5 and 7), the first and second visual areas (areas 17 and 18), area 21 of the temporal lobe, and areas of the orbital and lateral frontal cortex. Methods of fixation and immunocytochemical processing were designed to maximize the number of stained cells in 15-micron thick frozen sections and 1-micron-thick plastic sections. In 8 of the 10 areas the number and proportion of GABA-immunoreactive neurons per 50-micron-wide column were found to be the same (34-43 cells/column; 25% of the total neuronal population). Areas 17 and 3b differed. Area 17 contained 50% more GABA immunoreactive neurons (52-66 cells/column) but more than twice the total number of neurons, so that the GABA cells made up less than 20% of the total. In 3 monkeys, the number and proportion of GABA-positive neurons per 50-micron-wide column in area 3b were smaller than in adjacent areas of sensorimotor cortex (26 42 cells/column; 19-22%). In 2 other monkeys, the number and proportion (34-43 cells/column; 24-26%) were the same as in adjacent areas. Despite the similarity among most areas of monkey cortex, within some areas, the number of GABA-positive neurons per 50-micron-wide column varied as much as 30%. These variations form a significant, repeating pattern only in area 18, where narrow bands (150-200 micron wide) of relatively few stained cells alternated with either narrow or wide bands (600-700 micron wide) in which columns contained more cells. The GABA immunoreactive neurons were unevenly distributed across layers, with every area containing large numbers and proportions of stained cells in layer II, and every area but area 4 displaying a second concentration in the principal thalamocortical recipient layers. In area 4, the number of GABA-positive neurons declined sharply from layer II to layer III and remained low through layer VI. For areas displaying the greatest intra-areal variability, only 1 or 2 layers contributed significantly to that variability (layer IV in area 3b, layers III and V in area 18, and layers II and III in area 17).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3033171 TI - Potent and specific killing of human malignant brain tumor cells by an anti transferrin receptor antibody-ricin immunotoxin. AB - Immunotoxins are hybrid molecules which combine the exquisite selectivity of monoclonal antibodies with the potent toxicity of protein toxins. An immunotoxin was constructed by linking a murine monoclonal antibody against the human transferrin receptor (TR) to the plant toxin, ricin. The cytotoxic activity of the anti-TR-ricin immunotoxin was tested in vitro and demonstrated highly potent and cell type-specific killing of cells derived from human glioblastoma, medulloblastoma, and leukemia. The anti-TR-ricin immunotoxin killed more than 50% of "target" cells at a concentration of 5.6 X 10(-13) M after an 18-hour incubation with the ionophore, monensin. This potency exceeds that of any other anti-TR immunotoxin reported in the literature. When the activity of the anti-TR ricin immunotoxin against "target" tumor-derived cells was compared with the immunotoxin's activity against "non-target" cells, it could be predicted that a selective toxicity of anti-TR-ricin immunotoxin between tumor cells and normal brain was more than 150- to 1380-fold. Solid-phase indirect radioimmunoassay techniques were used to demonstrate significantly higher levels of TR in the glioblastoma- and medulloblastoma-derived cell lines, as well as in surgical tissue samples of medulloblastoma and glioblastoma, as compared to normal brain. Immunotoxins targeted to the TR may possess sufficient specificity to be of therapeutic importance, particularly to treat neoplastic disease of the central nervous system involving compartments (such as intrathecal, intraventricular, or cystic) where delivery of immunotoxins to tumor would not require transvascular transport. PMID- 3033172 TI - Imaging-based stereotaxic serial biopsies in untreated intracranial glial neoplasms. AB - Forty patients with previously untreated intracranial glial neoplasms underwent stereotaxic serial biopsies assisted by computerized tomography (CT) and magnetic resonance imaging (MRI). Tumor volumes defined by computer reconstruction of contrast enhancement and low-attenuation boundaries on CT and T1 and T2 prolongation on MRI revealed that tumor volumes defined by T2-weighted MRI scans were larger than those defined by low-attenuation or contrast enhancement on CT scans. Histological analysis of 195 biopsy specimens obtained from various locations within the volumes defined by CT and MRI revealed that: contrast enhancement most often corresponded to tumor tissue without intervening parenchyma; hypodensity corresponded to parenchyma infiltrated by isolated tumor cells or in some instances to tumor tissue in low-grade gliomas or to simple edema; and isolated tumor cell infiltration extended at least as far as T2 prolongation on magnetic resonance images. This information may be useful in planning surgical procedures and radiation therapy in patients with intracranial glial neoplasms. PMID- 3033173 TI - Autoradiographic comparison of thallium-201 diethyldithiocarbamate, isopropyliodoamphetamine and iodoantipyrine as cerebral blood flow tracers. AB - We investigated [201Tl]diethyldithiocarbamate (DDC) as a tracer for local cerebral blood flow (LCBF). Awake male rats were given intravenous infusions of a mixture of DDC and reference tracer(s): [123I]isopropyliodoamphetamine (IMP) and/or [14C]iodoantipyrine (IAP). LCBF values for DDC, IMP, and IAP were measured using simultaneous multiple radionuclide autoradiography and quantitative digital image analysis. Patterns of local cerebral blood flow (LCBF) obtained with DDC were intermediate compared to IMP and IAP, although they were more similar to those of IMP. DDC and IMP underestimated LCBF in some white matter and adjacent structures, while IAP underestimated LCBF values in high flow regions. We conclude that DDC uptake generally reflects local cerebral blood flow and that it can therefore be used as a cerebral perfusion tracer in humans using SPECT imaging. PMID- 3033175 TI - Ca2+ metabolism in arteries of spontaneously hypertensive rats: assessment by proton-induced X-ray emission. AB - Changes of intracellular Ca2+-metabolism have been considered to be of importance for the pathogenesis of essential hypertension. In the present study particle induced X-ray emission (PIXE) was used to get information on the spatial distribution of Ca2+ in aortas and renal arteries of spontaneously hypertensive (SHR) and normotensive rats. Our findings demonstrate a significant Ca2+ elevation in the aorta and renal artery of SHR versus normotensive Wistar-Kyoto (WKY) rats (3317 +/- 734 micrograms Ca2+/g tissue versus 1623 +/- 569 micrograms Ca2+/g tissue and 3432 +/- 1867 micrograms Ca2+/g tissue versus 1050 +/- 554 micrograms Ca2+/g tissue). PMID- 3033174 TI - Possible role of acetyl-CoA in the inhibition of CoA biosynthesis by ethanol in rats. AB - Ethanol, both administered to rats in vivo and added to cultured hepatocyte incubations, inhibits the conversion of [14C]pantothenate to coenzyme A (CoA). Data suggesting that the inhibition by ethanol involves its oxidation to acetate were obtained with rat hepatocytes maintained in primary culture. Ethanol, acetaldehyde and acetate were approximately equally effective inhibitors of [14C]pantothenate conversion to CoA (46-71%) and had no effect on uptake of [14C]pantothenate by hepatocytes. In the presence of saturating levels of acetate, acetaldehyde had no additional inhibitory effect. Cyanamide and diethyldithiocarbamate decreased the inhibition by acetaldehyde at the same concentration (10 microM), which saturated their ability to inhibit acetaldehyde oxidation. Studies with an isolated pantothenate kinase preparation showed that, of the ethanol metabolites, only acetyl-CoA was an effective inhibitor. Acetate and butyrate, which were both inhibitors of [14C]pantothenate conversion to CoA, increased the acetyl-CoA and decreased the free, unacylated CoA (CoASH) content of the cultured hepatocytes. The data were consistent with a mechanism for the inhibitory effect of ethanol that involves inhibition of pantothenate kinase by acetyl-CoA, but did not exclude a possible role of additional regulatory factors. PMID- 3033176 TI - The role of intraglomerular pressure in the initiation and progression of renal disease. AB - Reduction in functioning nephron number leads to progressive renal disease. A haemodynamic basis for this process has been suggested by studies of partially nephrectomized rats. In this model compensatory hyperfiltration in the remnant nephrons due to increases in the glomerular capillary hydraulic pressure (-PGC) and plasma flow rate is associated with eventual glomerular sclerosis. Therapeutic attenuation of these haemodynamic adaptations protects against glomerular injury. One such therapy is angiotensin converting enzyme (ACE) inhibition, which lowers systemic blood pressure and -PGC and prevents sclerosis in rats with renal ablation, as well as in the hyperfiltering kidneys of normotensive rats with diabetes mellitus. Control of -PGC with ACE inhibitor is also protective even when therapy is delayed until systemic hypertension and glomerular injury are established. In contrast, the control of systemic hypertension but not -PGC affords no protection in remnant kidney rats. Thus, control of glomerular hypertension slows the progression of renal disease. PMID- 3033177 TI - Reversing glomerular hypertension stabilizes established glomerular injury in renal ablation. AB - Male Munich-Wistar rats were studied 18 weeks after 1 2/3 nephrectomy. One group received no therapy. A second group received the angiotensin converting enzyme (ACE) inhibitor enalapril, starting 1 week after ablation. Two additional groups received no therapy during the first 8 weeks, and then received ACE inhibitor or a low (12%) protein diet. Early ACE inhibitor therapy resulted in control of systemic and glomerular hypertension (HTN), and a striking limitation of proteinuria and glomerular sclerosis. During the first 8 weeks untreated rats developed severe systemic HTN and increasing proteinuria. After 8 weeks proteinuria increased further in untreated rats, and widespread sclerosis resulted. Late ACE inhibition reversed systemic HTN. Both late ACE inhibition and late protein restriction reversed glomerular HTN and prevented further increases in proteinuria and sclerosis. Thus, control of glomerular HTN can stabilize renal injury even when therapy is delayed until hypertension and glomerular injury are established. PMID- 3033178 TI - Prevention of glomerular capillary hypertension in experimental diabetes mellitus obviates functional and structural glomerular injury. AB - Streptozotocin-diabetic rats kept moderately hyperglycaemic by daily injections of ultralente insulin for 4-6 weeks (group DM) demonstrated a higher glomerular transcapillary hydraulic pressure gradient (delta P), glomerular plasma flow rate and single-nephron glomerular filtration rate (GFR) than was observed in age- and weight-matched non-diabetic controls (group C). This rise in delta P was prevented by therapy with the angiotensin I converting enzyme inhibitor enalapril (15 mg/l drinking water, group DM + E) even though glomerular hyperperfusion and hyperfiltration persisted. Fourteen months after induction of diabetes, animals in group DM displayed high levels of albuminuria and an increased incidence of focal glomerular sclerosis; treatment with enalapril maintained these parameters at levels which did not differ from those observed in group C. We conclude that prevention of glomerular capillary hypertension with enalapril therapy obviates functional and structural glomerular injury in experimental diabetes mellitus. PMID- 3033179 TI - Effect of converting enzyme inhibitors on hypertensive large arteries in humans. AB - As demonstrated in hypertensive humans on the basis of non-invasive Doppler methods, converting enzyme inhibitors (CEI) dilate not only small arteries but also large arteries, particularly in the brachial and the carotid circulations. The dilating effect of CEI on large arteries is associated with an enhancement of systemic and brachial arterial compliance, implying a particular action of the drug on arterial smooth muscle. Converting enzyme inhibitors are thus able to reverse the reduced arterial compliance observed in patients with essential hypertension. PMID- 3033180 TI - Enalapril as a first-step agent in essential hypertension: a comparative study with atenolol. AB - The aim of the present double-blind crossover study was to compare the antihypertensive efficacy and tolerability of enalapril and atenolol in 48 patients with mild to moderate essential hypertension. After a 2-week wash-out period, treatment was started with either enalapril 20 mg or atenolol 50 mg daily. In patients with a diastolic blood pressure value of more than 90 mmHg after 2 weeks of therapy, doses were doubled. After 4 weeks of therapy, cases were classified as responders (diastolic blood pressure less than or equal to 95 mmHg) or non-responders (diastolic blood pressure greater than 95 mmHg) and after a 2-week wash-out phase switched over to the alternative drug for another 4-week therapy period. Both substances significantly lowered mean systolic and diastolic blood pressure to a comparable degree. After 2 weeks, 57% of patients under enalapril and 59% under atenolol shared a satisfactory response, which did not change at the higher dose levels. After 4 weeks of therapy the incidence of side effects was slightly, but insignificantly, higher on atenolol than on enalapril. Thus, our results show that both agents seem equally qualified as first-step drugs in the treatment of mild to moderate essential hypertension. PMID- 3033181 TI - Enalapril: a well-tolerated and efficacious agent for the paediatric hypertensive patient. AB - Enalapril, either alone or with a diuretic, was administered to 14 children with various renal diseases and hypertension. Five were transplant recipients on immunosuppressive agents. No adverse clinical or laboratory experiences were encountered. Renal function improved in seven children. Normal blood pressure for age was achieved in all 14 children. However, eight children required the addition of a diuretic. In four children blood pressure remained normal despite a reduction in the dose of enalapril. Enalapril was efficacious as a once per day agent. Concomitant ingestion of food did not attenuate the blood pressure lowering effect of enalapril. PMID- 3033182 TI - Different effects of two angiotensin converting enzyme inhibitors in primary hypertension--a comparison of captopril and enalapril. AB - A comparison of the antihypertensive effect and tolerability of captopril and enalapril was performed. Forty patients with primary hypertension were treated in randomized order, investigated immediately and in a crossover trial with treatment for 6 weeks, with an intermediate placebo wash-out of 3 weeks. A plasma renin profile was obtained from each patient before treatment. The immediate maximal mean change in supine blood pressure (BP) was similar with both drugs, but occurred earlier with captopril. After 6 weeks of treatment enalapril was more potent than captopril in the mean reduction of supine BP produced. This was even more evident in patients with low-renin hypertension (LRH), where captopril had practically no effect. The different chronic BP response in relation to initial renin activity may indicate different mechanisms or modalities of action. Following the recommended doses and administration intervals of captopril and enalapril, the latter seems more effective, but there seems to be no difference in tolerability. PMID- 3033184 TI - Acute hypotensive effect of calcium antagonists and endogenous digitalis-like immunoreactivity in human essential hypertension. AB - Evidence has been provided on the increased presence, in essential hypertension, of endogenous digitalis-like factor(s) [DLIS, digoxin-like immunoreactive substance(s)] able to cross-react with antidigoxin antibodies and to inhibit the membrane-bound sodium-potassium pump. An inhibition of the sodium pump could lead, in smooth muscle cells, to an increase of intracellular calcium ions and to an increase of total peripheral resistances. In this study the relation between plasma levels of DLIS and the acute hypotensive effect of a calcium antagonist (nifedipine) has been evaluated in a group of borderline to severe hypertensive patients and in a control group of normotensive subjects. The results obtained confirm that the hypotensive effect of nifedipine is related to pretreatment blood pressure and show, only in hypertensive patients, a significant relation of DLIS with both pretreatment blood pressure and blood pressure decrement induced by nifedipine. These findings are compatible with a possible role of DLIS in modulating cellular calcium handling. PMID- 3033183 TI - Angiotensin converting enzyme inhibition and calcium channel blockade as primary antihypertensive therapy. AB - Recent large-scale antihypertensive treatment trials primarily emphasize the quality of blood pressure control for reduction of cerebrovascular accidents as well as for myocardial infarction, practically irrespective of the type of drug used. Therefore, the best drug that normalizes blood pressure without adverse effects should be sought. On the basis of studies demonstrating cellular membrane and calcium homoeostatic derangements, and an age-dependent transition of overall cardiovascular regulation and peripheral vasoconstrictor forces during the course of essential hypertension, an alternative treatment concept is proposed: angiotensin converting enzyme inhibitors or beta-blockers can primarily be used in younger patients and those with a high renin, while calcium antagonists are used in place of diuretics in older, low-renin or black patients. Age-oriented two-way drug selection enables normalization of blood pressure without untoward effects in about 80% of patients with essential hypertension, and helps to optimize drug combinations in those patients who are difficult to treat. PMID- 3033185 TI - Modification of stained enamel surfaces: use of hydrochloric acid and pumice mixture. PMID- 3033186 TI - Evaluation of collagen/hydroxylapatite for augmenting deficient alveolar ridges: a preliminary report. AB - A multicenter study was undertaken to evaluate a new alveolar ridge augmentation material composed of purified fibrillar collagen and particulate hydroxylapatite (PFC/HA). In a study of 77 patients and 99 reconstructed ridges, this material provided superior handling properties over HA alone as evidenced by its ease of surgical placement and manipulation, and the rarity of particle migration or displacement. Moreover, the rapid development of ridge firmness and stability allowed for the loading of dentures within three to six weeks. Prosthodontist evaluations and surveys of patient satisfaction showed great satisfaction with denture fit, comfort, esthetics, speech, and ability to masticate. PMID- 3033187 TI - Mandibular augmentation in dogs with hydroxylapatite combined with demineralized bone. AB - This study investigates the tissue response in dogs to mandibular augmentation with hydroxylapatite (HA) combined with demineralized bone. Thirty mongrel dogs underwent ridge augmentation with 3 ml of HA alone (HAO), HA combined with autogenous bone (HAB), or HA combined with demineralized bone (HAD). Bone was not found in the HAO augmented ridges through 52 weeks; bone was found in the HAB augmented ridges by 12 weeks, and the HAD augmented ridges had bone formation after 26 weeks. This study demonstrates that demineralized bone can induce osteogenesis within the HA augmented dog ridge, but it is delayed when compared to HAB augmentations. PMID- 3033188 TI - Porous hydroxylapatite as a bone graft substitute in mandibular contour augmentation: a histometric study. AB - The buccal contour of the mandible was augmented in 17 dogs with 5 X 7.5 X 20 mm blocks of porous hydroxylapatite (HA) on one side and two-layered split rib autografts on the other. Both specimens were retrieved at three, six, 12, 24, and 48 months. Undecalcified sections were prepared for microradiography, light and UV microscopy, and histometry. A transmitted light video image digitizing system was used to trace implant and graft perimeters and calculate cross sectional areas. This system was also used to measure graft density and calculate bone and soft tissue compositions. The HA matrix, bone and soft tissue compositions of implant specimens were measured with a backscattered scanning electron microscope imaging digitizing system. All grafts became increasingly resorbed with time whereas all implants remained intact. Mature osteotonic bone ingrowth was present in all implants except one which failed to unite with the mandibular cortex. The mean graft areas decreased from 30.8 mm2 at three months to 0.7 mm2 at 48 months, while the implant areas averaged 35.5 mm2 and remained stable. The graft specimens were composed of 46.6% bone and 53.4% soft tissue or fluid space. The implant specimens were composed of 34.5% HA matrix, 28.6% bone, and 33.9% soft tissue. The HA matrix had a surface area of 9.8 mm2/mm3 that was 61.9% covered with bone ingrowth and 38.1% covered with soft tissue or fluid space. In contrast to the rapid resorption of graft onlays, the porous HA matrix demonstrated a long term permanence with maintenance of contour and osseous incorporation over the four-year duration of this study. PMID- 3033189 TI - Elastosis in breast carcinoma: I. Immunohistochemical characterization of elastic fibres. AB - Elastosis associated with invasive ductal and lobular carcinomas of the breast was examined by tinctorial and immunohistochemical staining methods, enzyme digestion, and electron microscopy. The elastotic material exhibited the tinctorial staining properties of elastic fibres, and the ultrastructural appearances were those of elastic fibres although there was a higher proportion of microfibrils than in normal mature elastic fibres. The elastosis was immunostained by antisera to human fetal elastin, lysozyme and amyloid P component, as in other sites where elastic fibres are found. These findings indicate that immunohistochemically intact elastic fibres are present in the elastosis of breast cancer. They also demonstrate that lysozyme and amyloid P component are co-distributed with elastic fibres in elastosis of breast carcinoma, as distinct components with different susceptibilities to enzyme digestion. The cellular origin of elastosis in breast carcinoma remains uncertain. PMID- 3033190 TI - Effects of ketoconazole and itraconazole on growth and sterol synthesis in Pityrosporum ovale. AB - The effects of ketoconazole and itraconazole on growth and sterolsynthesis in Pityrosporum ovale was studied. Itraconazole was at least 10 times more active than ketoconazole. Sterol synthesis was inhibited more rapidly than growth, suggesting that the antifungal activity of both azoles originates from an effect on the 14 alpha demethylase system, as seen in other species. PMID- 3033191 TI - Cavitron assisted liver resection in a child. AB - Resection of a liver tumor using the cavitron ultrasound dissector is described. The technique is safe and minimizes blood loss. PMID- 3033192 TI - Role of dopaminergic and GABAergic mechanisms in discrete brain areas in phencyclidine-induced locomotor stimulation and turning behavior. AB - This study was designed to test whether phencyclidine (PCP)-induced turning behavior and locomotor stimulation result from the action of this drug on functionally different neuronal systems and different sites of the brain. PCP produced turning behavior towards the drug injection side with unilateral injection of PCP (50-100 micrograms) into the globus pallidus, but not the nucleus accumbens and the caudate nucleus. This turning behavior was strongly attenuated by a gamma-aminobutyric acid (GABA) antagonist, bicuculline, and by pimozide which reduces dopaminergic transmission in non-injection sites. Turning behavior induced by intraperitoneal injection of PCP (7.5 mg/kg) was enhanced by a GABA agonist, baclofen, and attenuated by GABA antagonists (bicuculline, picrotoxin). On the other hand, PCP produced significant locomotor stimulation, sniffing, rearing and forward locomotion with unilateral injection of 25-100 micrograms into the nucleus accumbens and the caudate nucleus. These behaviors were strongly antagonized by intraperitoneal injection of pimozide. The locomotor stimulation induced by intraperitoneal injection of PCP (5 mg/kg) was markedly enhanced by a small dose of methamphetamine and, by contrast, attenuated by reserpine, 6-hydroxydopamine, haloperidol, pimozide and a low dose of apomorphine which inhibits the release of dopamine by the stimulation of presynaptic receptors. These results suggest that PCP-induced turning behavior may be produced through stimulation of GABAergic transmission in the globus pallidus, although PCP-induced locomotor stimulation, sniffing, rearing and forward locomotion may be produced by increasing dopaminergic transmission in the nucleus accumbens and the caudate nucleus. PMID- 3033193 TI - [Autonomously replicating sequence activity of kinetoplast DNA in yeast cells]. AB - Different fragments of the maxicircle of the kinetoplast DNA (kpDNA) from Crithidia oncopelti were cloned (Fig. 1, 2) and tested for ARS-activity (ARS, autonomously replicating sequences). ARS-activity was expressed as number of transformed yeast cells per microgram plasmid DNA, as number of transformed cells per number of plasmid molecules (transformation efficiency, TE) and as number of transformed cells per kilobase pair of cloned kpDNA (specific transformation efficiency, TES) (Fig. 3, Table 1). All DNA fragments studied showed ARS activity. Large fragments exhibited higher ARS-activities than their smaller subfragments. The number of fragments showing ARS activity and their distribution within the maxicircles (Fig. 4) suggest that there was no strong correlation between sites with ARS-activity and the replication origin of kpDNA. PMID- 3033194 TI - Spread of plasmid-mediated nourseothricin resistance due to antibiotic use in animal husbandry. AB - After using of the streptothricin antibiotic nourseothricin in animal husbandry for growth promotion, plasmid-borne resistance to streptothricin could be observed in E. coli from nourseothricin fed pigs, from employees in pig farms and from their family members. Moreover, streptothricin resistance plasmids also occurred in E. coli of man without any contact to pig farms (gut flora and even urinary tract infections). However, these individuals live in villages and towns of the territory where nourseothricin was applied to pigs. Similar streptothricin resistance plasmids belonging to different incompatibility groups were found in both E. coli from pigs and E. coli from human beings. As no coselection of resistance to drugs indispensable for therapeutic use in man was observed, the application of nourseothricin in animal husbandry has not clinical implication for human medicine yet. Nevertheless, this problem remains under further investigation. PMID- 3033195 TI - Synthesis and antiviral activity of various esters of 9-[(1,3-dihydroxy-2 propoxy)methyl]guanine. AB - The synthesis and in vivo biological activity of a series of mono-O-, di-O-, and N2-acyl derivatives of 9-[(1,3-dihydro-2-propoxy)-methyl]guanine (DHPG) are described. PMID- 3033196 TI - Purification and characterization of the plantar human papilloma virus. PMID- 3033197 TI - Subungual fibrous histiocytoma mimicking melanoma. PMID- 3033198 TI - Salutary effects of prostaglandin E1 in perfused rat lungs injured with hydrogen peroxide. AB - Studies were conducted in isolated, buffer-perfused rat lungs to determine if prostaglandin (PG) E1 attenuated pulmonary edema provoked by hydrogen peroxide (H2O2). When lungs were challenged by 60 min of perfusion with H2O2 (generated by the reaction between glucose and glucose oxidase) the wet weight-to-dry weight ratio increased from control by 54%, indicating development of pulmonary edema. In contrast, lungs treated simultaneously with H2O2 plus PGE1 (1 microgram/min) failed to exhibit an elevated wet-to-dry weight ratio. H2O2-injured lungs demonstrated a modest 2 torr increase in pulmonary arterial perfusion pressure that was not influenced by simultaneous treatment with PGE1. Both radioimmunoassay (RIA) and high-performance liquid chromatographic (HPLC) analysis detected increased amounts of (5S)-5-hydroxy-6,8,11,14 eicosatetraenoic acid in the perfusion medium of H2O2-injured lungs (RIA, 48.0 +/- 14.7; HPLC, 54.8 +/- 13.5) relative to controls (RIA, 6.6 +/- 1.6; HPLC, 6.8 +/- 1.9), and simultaneous treatment with PGE1 tended to blunt this increase (RIA, 29.2 +/- 8.3; HPLC, 29.8 +/- 7.6). PGE1 abolished the increase in wet weight-to-dry weight ratio induced by exogenous leukotriene C4. Production of H2O2 by the glucose glucose oxidase reaction was not influenced by PGE1. Taken together, these observations indicate that PGE1 attenuates H2O2-induced pulmonary edema formation in buffer-perfused rat lungs by mechanisms that may relate to inhibition of lung 5'-lipoxygenase activation and/or to inhibition of the injurious effects of endogenously produced lipoxygenase products. PMID- 3033199 TI - Molecular mechanisms of corticotropin-releasing factor stimulation of calcium mobilization and adrenocorticotropin release from anterior pituitary tumor cells. AB - In a tumor cell line of the mouse anterior pituitary (AtT-20/D16-16) consisting of a homogeneous population of corticotrophs, corticotropin-releasing factor (CRF) activates adenylate cyclase and cAMP-dependent protein kinase. In addition, CRF induces a rise in cytosolic calcium levels in AtT-20/D16-16 cells and stimulates adrenocorticotropin hormone release. To determine whether activation of cAMP-dependent protein kinase is essential for CRF to stimulate calcium mobilization and trigger adrenocorticotropin hormone release, an inhibitor of cAMP-dependent protein kinase was inserted into AtT-20/D16-16 cells using a liposome technique. In control cells, CRF, forskolin (a direct activator of adenylate cyclase) and potassium increased cytosolic calcium levels. Insertion of the protein kinase inhibitor into AtT-20/D16-16 cells greatly attenuated CRF and forskolin-stimulated calcium mobilization although it did not alter the rise in cytosolic calcium induced by potassium. Treatment of the cells with liposomes lacking protein kinase inhibitor (but containing an equivalent amount of bovine serum albumin) had no effect upon the calcium mobilization elicited by any of the agents tested. These results reveal an essential role for cAMP-dependent protein kinase in mediating CRF-stimulated calcium mobilization and suggest that its activation may be an essential molecular event for CRF to evoke adrenocorticotropin hormone secretion. PMID- 3033200 TI - Effects of clenbuterol on central beta-1 and beta-2 adrenergic receptors of the rat. AB - Repeated administration of the centrally acting beta adrenoceptor agonist, clenbuterol, to rats reduced the ability of isoproterenol to increase the concentration of cyclic AMP (cAMP) in slices of cerebellum. This reduced responsiveness to isoproterenol was accompanied by a marked reduction in the density of beta adrenoceptors as measured by the binding of the beta adrenoceptor antagonist [125I]iodopindolol. In addition, the agonist-binding properties of remaining cerebellar beta adrenoceptors were altered after clenbuterol treatment. The clenbuterol-induced reduction in the density of beta adrenoceptors in the cerebellum is in marked contrast to its inability to do this in cerebral cortex. Comparison of the ability of clenbuterol to that of isoproterenol to increase levels of cAMP in slices of cerebral cortex or cerebellum showed that clenbuterol is a weakly potent agonist in both brain regions. The increase in cAMP induced by isoproterenol in the cortex was significantly reduced in the presence of the selective beta-1 adrenoceptor antagonist, ICI 89,406. In contrast, the clenbuterol-induced increase in cortical cAMP was unchanged by ICI 89,406 but was reduced significantly by the beta-2 adrenoceptor antagonist, ICI 118,551. In cerebellum, both isoproterenol- and clenbuterol-stimulated accumulation of cAMP were antagonized much more potently by ICI 118,551 than by ICI 89,406. Furthermore, clenbuterol antagonized the cAMP response induced by isoproterenol in the presence of ICI 118,551 in a concentration-dependent manner. In terms of measurement of cAMP in brain slices, clenbuterol is weakly potent as an agonist at beta-2 adrenoceptors and has antagonist properties at beta-1 adrenoceptors. PMID- 3033201 TI - Vasoactive intestinal peptide and intraocular pressure: adenylate cyclase activation and binding sites for vasoactive intestinal peptide in membranes of ocular ciliary processes. AB - Vasoactive intestinal peptide (VIP)-responsive adenylate cyclase and VIP binding sites were investigated in membranes prepared from ciliary processes dissected from albino rabbit eyes. High-affinity binding sites for VIP (Kd, 0.95 nM; 607 fmol/mg of protein), in addition to beta adrenergic sites labeled by dihydroalprenolol (Kd, 0.48 nM; 123 fmol/mg of protein), were present. Activation of adenylate cyclase by VIP had a Ka of 65 nM, and the maximal response was 3.3 fold greater than that for I-isoproterenol (Ka, 102 nM). A peptide fragment of VIP (sequence 10-28) was inactive in all assays and did not inhibit VIP stimulated adenylate cyclase at 10 microM. Responses to VIP and isoproterenol in combination were additive at lower doses but less than additive at maximal doses. Responses to VIP in combination with a low dose of forskolin (0.1 microM) were potentiated at all dose levels, whether assays were done in presence of MgCl2 or MnCl2. VIP- and forskolin-activated adenylate cyclase was associated with the nonpigmented epithelial cell fraction and not with pigmented epithelial cells separated on Percoll density gradients after dissociation of cells from processes by collagenase digestion. Intravitreous injection of 10 nmol of VIP into the rabbit eye caused a maximal reduction in intraocular pressure at 40 to 50 hr lasting beyond 72 hr. VIP-responsive and beta adrenergic-responsive adenylate cyclase are present on the same cell type (nonpigmented epithelial cells) and appear to share components of the adenylate cyclase system in the same membrane. VIP may participate in the physiologic regulation of aqueous humor secretion at the level of the epithelial cell membrane. PMID- 3033202 TI - Variation in sensitivity of alpha adrenoceptor-mediated contraction of the vascular smooth muscle of rabbit elastic and muscular arteries is related to receptor affinity. AB - Norepinephrine sensitivity (pD2) and agonist dissociation constant (pKA) have been determined in the following 12 rabbit arteries: thoracic and abdominal aorta, basilar, ear, common, external and internal iliac, ovarian, large and medium pulmonary, renal and superior mesenteric. They were determined in the presence of beta adrenoceptor blockade and uptake 1 and uptake 2 inhibition to prevent compromising additional actions of norepinephrine and intrinsic processes that influence its concentration at the site of action. In the superior mesenteric artery, determinations were made after endothelial inactivation and in the presence of indomethacin, because in this vessel blockade by these procedures influences norepinephrine sensitivity. In 12 arteries a positive correlation was found between norepinephrine pD2 and pKA (r = 0.74, P less than .01). The slope of the regression line did not differ from unity. Norepinephrine pD2 did not correlate with receptor reserve in these arteries when assessed as antilog pD2 pKA. In three arteries, the ear and common and external iliac, a large receptor reserve was found. If these were excluded from the series, the following correlation would be found: r = 0.9; P less than .001. Here again the slope of the regression line did not differ from unity. The pD2 and pKA were determined for the more selective alpha-1 adrenoceptor agonist, phenylephrine in six of these arteries, and similar results were obtained. The KB for prazosin in this series did not correlate with norepinephrine KA (r = 0.45, P greater than .05), and the slope (0.17) was not significantly different from zero. The pD2 for histamine, determined after H2 receptor blockade, does not differ in the arteries.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3033203 TI - Fengabine, a novel antidepressant GABAergic agent. I. Activity in models for antidepressant drugs and psychopharmacological profile. AB - Fengabine (SL 79.229) is a novel benzylidene derivative with clinically proven antidepressant action. Fengabine is active in behavioral models for antidepressant drug action, reversing the passive avoidance deficit in olfactory bulbectomized rats, antagonizing the escape deficit in the learned helplessness model and decreasing paradoxical sleep in the rat. In contrast to tricyclic antidepressants, fengabine antagonizes 5-hydroxytryptophan-induced head twitches and only weakly reverses reserpine-induced ptosis. Fengabine inhibits neither monoamine uptake nor monoamine oxidase. A GABAergic mechanism of fengabine is indicated as bicuculline reverses its action in the olfactory bulbectomy and learned helplessness models. The wide-spectrum anticonvulsant action of fengabine is consistent with a GABA-mimetic action and is in contrast to the proconvulsant effect of most classical antidepressants. PMID- 3033204 TI - Fengabine, a novel antidepressant GABAergic agent. II. Effect on cerebral noradrenergic, serotonergic and GABAergic transmission in the rat. AB - The effects of fengabine (a novel benzylidene derivative possessing clinically demonstrated antidepressant action) on neurochemical parameters related to norepinephrine, serotonin and gamma-aminobutyric acid (GABA) neurons have been investigated in the rat and mouse brain. When given acutely, fengabine (50-1000 mg/kg i.p.) does not alter norepinephrine uptake but accelerates the turnover rate of norepinephrine in the rat brain as demonstrated by the enhancement of: the alpha-methyl-p-tyrosine-induced disappearance of norepinephrine in the hypothalamus; 3,4-dihydroxyphenylacetic acid levels in noradrenergic cell body areas; the pargyline-induced accumulation of normetanephrine in the hypothalamus; and 3,4-dihydroxyphenylethyleneglycol levels in the hypothalamus, septum and spinal cord. No tolerance to the effect of fengabine on the latter biochemical parameter was observed after repeated treatment for 2 weeks at doses of 100 or 200 mg/kg i.p., b.i.d. Fengabine (100 or 200 mg/kg i.p., b.i.d.), given for 14 days, causes a desensitization of isoprenaline-stimulated adenylate cyclase in septal and cortical slices of the rat but fails to modify cortical beta, alpha-1 or alpha-2 adrenoceptor binding sites. Fengabine (up to 400 mg/kg i.p.) has no effect on rat cerebral serotonin uptake, synthesis or metabolism. Moreover, when given subacutely (100 or 200 mg/kg i.p., b.i.d. for 2 weeks), it fails to alter rat cortical serotonine receptors or [3H]imipramine binding sites. Fengabine (up to 50-100 microM) is also inactive in vitro on [3H] GABA binding to GABAA or GABAB receptors in the rat brain or on GABA transaminase activity in the mouse brain.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3033205 TI - Beta-2 adrenoceptor-mediated relaxation of the isolated human saphenous vein. AB - On isolated strips of human saphenous vein, pretreated with 5 microM phenoxybenzamine and contracted with 10 mM KCl, the beta adrenoceptor mediating the relaxant effects of isoproterenol, procaterol and norepinephrine was characterized using the selective beta-1 adrenoceptor antagonist, bisoprolol, and the selective beta-2 adrenoceptor antagonist, ICI 118,551. All three agonists produced concentration-dependent relaxations of the isolated saphenous vein with an order of potency: procaterol (pD2 value, 7.69) greater than isoproterenol (pD2 value, 7.41) much greater than norepinephrine (pD2 value, 5.30). ICI 118,551 (3 X 10(-10) to 3 X 10(-9) M) was nearly 100 times more potent than bisoprolol (10(-7) to 10(-6) M) in antagonizing the relaxant effects of isoproterenol and procaterol. The slopes of the Schild plots for the antagonistic effects of ICI 118,551 and bisoprolol against isoproterenol- and procaterol-induced relaxations were not significantly different from unity indicating interaction with a homogeneous population of beta adrenoceptors. The pA2 value for ICI 118,551 amounted to 9.11 to 9.20 and for bisoprolol to 6.50 to 6.63. In addition, the concentration-response curve for the relaxant effect of norepinephrine was significantly shifted to the right by 10(-9) M ICI 118,551, but not affected by 10(-7) M bisoprolol. These results indicate that on the isolated strips of the human saphenous vein the beta adrenoceptor mediating relaxation is of the beta-2 subtype. PMID- 3033206 TI - Nitrite conversion to nitric oxide in red cells and its stabilization as a nitrosylated valency hybrid of hemoglobin. AB - The authors describe a method for the preparation, isolation and purification from human red blood cells of a stable form of hemoglobin containing both oxidized and reduced subunits. After isoelectric focusing across a pH gradient, it occupies the same position as the synthetically reconstituted or chemically generated species, (alpha 2+ beta 3+)2 (I). Unlike its synthetic or chemically generated counterpart, however, it (HbX) does not bind oxygen. A different method for the preparation in situ of the (alpha 3+ beta 2+)2 valency hybrid results in a pigment with chemical and spectral properties identical with those ascribed to its synthetically reconstituted counterpart, and both bind oxygen. HbX is generated in highest yield when red cells are incubated under N2 in the presence of glucose, methylene blue and nitrite for several hours. Its visible absorption spectrum differs from that reported for I, and it reacted very slowly with ferricyanide. Exposure to CO did not result in spectral shifts over that for HbX in air, but spectral shifts were produced with CO exposure of (alpha 3+ beta 2+)2. HbX had an inositol hexaphosphate difference-binding spectrum quite unlike that described for I. HbX also had a distinctive electron paramagnetic resonance spectrum that shifted after inositol hexaphosphate addition with a new three-line hyperfine pattern. Both spectra were characteristic for an unpaired electron in an iron-centered orbital with a hyperfine coupling to the nitrogen nuclear spin of a nitrosyl ligand on heme iron. The authors conclude that HbX is a form of I, but it has NO bound to its reduced subunits. PMID- 3033207 TI - Interactions of metaphit with phencyclidine and sigma agonist actions in rat cerebellum: determination of specificity and selectivity. AB - The interactions of phencyclidine (PCP) and related agonists with putative receptor blockers were studied on cerebellar Purkinje neurons using electrophysiological techniques. Depressions induced by PCP or dexoxadrol, a sigma receptor agonist, were markedly antagonized by the PCP receptor antagonist metaphit, which acylates PCP receptors via its isothiocyanate moiety. Conversely, the depressant effect of levoxadrol, the (-) isomer of dexoxadrol, was not affected by metaphit. Further evidence that metaphit's specific antagonism of dexoxadrol- and PCP-mediated depressions was derived from data showing that drugs which respectively acylate mu and delta opioid receptors, benzimidazole isothiocyanate and fentanyl isothiocyanate, do not antagonize the actions of either PCP or dexoxadrol. Moreover, tyramine, which like PCP acts as an indirect norepinephrine agonist, is not antagonized by metaphit. These observations support the concept that metaphit causes a pharmacologically specific and irreversible antagonism of the effects of both PCP and dexoxadrol in the cerebellum. Thus, the electrophysiological mechanisms of PCP actions are similar to those triggered by sigma opioid agonists in this brain area. PMID- 3033209 TI - Structure-activity relationships for steroid interaction with the gamma aminobutyric acidA receptor complex. AB - Certain steroids are potent barbiturate-like modulators of the gamma-aminobutyric acidA (GABA) receptor-chloride ionophore complex in rat brain membranes. At nanomolar to low micromolar concentrations, these steroids stimulate [3H]flunitrazepam and [3H] muscimol binding and displace the convulsant [35S]t butylbicyclophosphorothionate from its binding site in an allosteric manner, in addition to enhancing Cl- conductance responses to GABA recorded in cultured rat hippocampal and spinal neurons. A stringent structure-activity relationship exists for these interactions of steroids with the GABAA receptor complex. Comparison of the structure-activity relationship data obtained in this study with those for steroid-induced general anesthesia strongly suggests that steroidal anesthesia may result from the interaction between steroids and the GABAA receptor. The essential features of the active structures are a 5 alpha or 5 beta-reduced pregnane skeleton with a hydroxyl at C3 in the alpha-position and a ketone group at C20. These features are all present in some naturally occurring steroids, including metabolites of deoxycorticosterone and progesterone, that show potent activity at the GABAA receptor complex. Two of the compounds investigated are known to be formed in vivo as reduced metabolites of endogenous steroid hormones: 5 alpha-pregnane-3 alpha -ol-20-one and 5 alpha-pregnane-3 alpha,21-diol-20-one, which are derived from progesterone and deoxycorticosterone, respectively. These two steroids produce a striking prolongation of GABA-mediated inhibitory postsynaptic currents recorded at synapses between rat hippocampal neurons in culture and could conceivably regulate GABA-mediated inhibition under some physiologic and pathologic conditions. PMID- 3033210 TI - Comparison of the alpha adrenoceptor activity of dopamine, ibopamine and epinine in the pulmonary circulation of the dog. AB - The ability of dopamine, ibopamine and epinine to elicit alpha adrenoceptor mediated vasoconstriction was studied in the in situ, autoperfused pulmonary circulation of the open-chest anesthetized dog. Animals were pretreated with propranolol to eliminate beta adrenoceptor-mediated relaxation of the pulmonary vasculature. Heparinized blood was withdrawn from the left femoral artery and transferred via a peristaltic pump to the pulmonary arterial branch supplying the left diaphragmatic lobe of the lung. The flow rate of the pump was adjusted so that mean pulmonary perfusion pressure in the lobe was equal to resting diastolic pulmonary artery pressure (10 +/- 1 mm Hg). Under conditions of constant left atrial pressure and pulmonary blood flow, intralobar administration of dopamine, ibopamine and epinine elicited dose-dependent increases in perfusion pressure of the lobe, reflecting increases in pulmonary vascular resistance. Prazosin (100 micrograms/kg i.v.), a selective alpha-1 adrenoceptor antagonist, inhibited the pulmonary vasopressor responses to dopamine, ibopamine and epinine. Rauwolscine (100 micrograms/kg i.v.), a selective alpha-2 adrenoceptor antagonist, inhibited pulmonary pressor responses to dopamine and epinine without altering significantly the pulmonary vasoconstrictor response to ibopamine. These data indicate that dopamine and epinine stimulate both postjunctional vascular alpha-1 and alpha-2 adrenoceptors to elicit pulmonary vasoconstriction in the dog, whereas ibopamine, when injected directly into the pulmonary circulation, stimulates primarily postjunctional vascular alpha-1 adrenoceptors. However, when ibopamine was administered intraduodenally, both prazosin and rauwolscine were found to inhibit the resulting pulmonary vasopressor response. This finding is consistent with the hypothesis that ibopamine is converted to its active metabolite epinine, which stimulates both pulmonary vascular alpha-1 and alpha-2 adrenoceptors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3033208 TI - Mu opioid receptor involvement in enkephalin activation of dopamine neurons in the ventral tegmental area. AB - Many lines of evidence suggest that opioids act in the A10 dopamine (DA) region to activate DA neurons projecting to limbic terminal areas. Thus, injection of morphine and enkephalin analogs into the ventral tegmental area (a major subnucleus of the A10 DA region) produces an increase in spontaneous motor activity that is blocked by DA receptor antagonists and increases DA metabolism in the nucleus accumbens. The present study utilized enkephalin analogs specific for either the mu or delta opioid receptor to evaluate which receptor subtype(s) is activating the A10 DA neurons. It was found that the specific mu agonist, Try D-Ala-Gly-NMe-Phe-Gly-ol, was significantly more potent than the specific delta agonist, [D-Pen2,5]-enkephalin, at increasing spontaneous motor activity or DA metabolism in the nucleus accumbens, septum, striatum and prefrontal cortex. Further, naloxonazine, a putative antagonist of the mu-1 isoreceptor, significantly attenuated the motor-stimulant effect and increase in DA metabolism produced by intra-ventral tegmental area injection of Tyr-D-Ala-Gly-NMe-Phe-Gly ol. It was found that the disposition of microinjected Tyr-D-Ala-Gly-NMe-Phe-Gly ol or [D-Pen2,5]-enkephalin was not responsible for the difference in their potency. It is concluded that the mu receptor and, perhaps, the mu-1 isoreceptor mediate a major portion of the activation of A10 DA neurons previously demonstrated with mixed mu and delta opioid agonists. PMID- 3033211 TI - Human postjunctional alpha-1 and alpha-2 adrenoceptors: differential distribution in arteries of the limbs. AB - Experiments were performed in order to characterize the post-junctional alpha adrenoceptors that mediate contraction in arteries of human limbs. Blood vessels were obtained from patients undergoing amputation of an extremity for reasons other than vascular disease. Proximal (dorsalis pedis and arcuate arteries of the foot, superficial palmer arch of the hand) and distal (digital arteries of the foot and hand) blood vessels were studied from each limb. The blood vessels were removed within 60 min of amputation and were suspended for isometric tension recording in modified Krebs-Ringer bicarbonate solution. In proximal and distal arteries, alpha-1 adrenergic blockade with prazosin produced a nonparallel shift in the concentration-effect curve to high compared to low concentrations of the agonist. In contrast, alpha-2 adrenergic blockade with rauwolscine was more effective against responses evoked by low concentrations of norepinephrine. This suggests that the alpha-2 adrenergic component of the response to norepinephrine is a low-maximum effect compared to the alpha-1 adrenergic component. Prazosin was less potent and rauwolscine more potent in distal arteries, compared to proximal arteries which might indicate an increased alpha-2 adrenergic response in distal arteries. The selective alpha-1 adrenergic agonist, phenylephrine, produced similar responses in proximal and distal arteries. However, the selective alpha-2 adrenergic agonist, B-HT 920, caused greater contractile responses in distal arteries compared to proximal arteries. The results suggest that alpha-1 and alpha-2 adrenoceptors are present on the vascular smooth muscle of arteries of human limbs, and that alpha-2 adrenoceptors are more prominent on distal arteries. This may be related to an increased contribution of the distal arteries to thermoregulation. PMID- 3033212 TI - Phorbol ester-induced inhibition of cyclic GMP formation mediated by muscarinic receptors in murine neuroblastoma cells. AB - The effects of phorbol 12-myristate 13-acetate (PMA) on carbamylcholine (CBC) induced [3H]cyclic GMP formation in mouse neuroblastoma cells (clone N1E-115) were studied. PMA, but not 4 alpha-phorbol, suppressed muscarinic receptor mediated cyclic GMP responses in a time-dependent and a concentration-dependent fashion with an IC50 of 68.8 +/- 20.2 nM. The inhibitory effects of PMA on CBC induced cyclic GMP formation were of a mixed competitive and noncompetitive type, being characterized by a depression of maximal cyclic GMP response to CBC and a significant increase in its EC50. PMA also significantly reduced [3H]cyclic GMP formation induced by histamine, without affecting the responses elicited either by sodium azide or the calcium ionophore A23187. Although the inhibitory effects of PMA on CBC-induced cyclic GMP formation were not reversed by washing, these effects were significantly attenuated by H-7 [1-(5-isoquinolinesulfonyl)-2 methylpiperazine], a protein kinase C inhibitor. PMA had no effect on binding of an antagonist ligand to muscarinic receptors, or on the binding characteristics of CBC to these receptors in intact cells. On the other hand, PMA competed for the specific binding of a labeled phorbol ester in intact cells with a potency similar to that of PMA in inhibiting muscarinic receptor-mediated [3H]cyclic GMP responses. PMID- 3033214 TI - Role of mu and delta receptors in the supraspinal and spinal analgesic effects of [D-Pen2, D-Pen5]enkephalin in the mouse. AB - The opioid receptors involved in the supraspinal and spinal actions of [D-Pen2, D Pen5]enkephalin (DPDPE) for production and/or modulation of analgesia were investigated in two thermal analgesic tests, the mouse warm water (55 degrees C) tail-withdrawal assay and the radiant heat tail-flick test. Two approaches were used at supraspinal and spinal sites: determination of possible cross-tolerance between morphine and a variety of receptor selective/nonselective agonists (DPDPE, [D-Pen2, L-Pen5]enkephalin (DPLPE), [D-Ala2, MePhe4, Gly-ol]enkephalin, [D-Ala2, Met5]enkephalin amide, [D-Ser2, Leu5, Thr6]enkephalin and [D-Thr2 Leu, Thr6]enkephalin) and possible potentiation of morphine (mu) analgesia by proposed delta agonists (DPDPE, DPLPE and [D-Ala2, D-Leu5]enkephalin) in naive and morphine-tolerant mice. Additionally, proposed mu (morphine) and delta (DPDPE) agonists were evaluated for their i.c.v. analgesic effectiveness in the absence, and in the presence, of the proposed delta antagonist ICI 174,864. The present communication now reports that after i.c.v. administration analgesic cross tolerance could be demonstrated between morphine and a variety of relatively selective or nonselective opioids but not to the highly delta selective DPDPE and DPLPE. This result was consistent with direct antagonism of i.c.v. DPDPE, but not morphine analgesia, by ICI 174,864. Furthermore, i.c.v. DPDPE and DPLPE were able to potentiate morphine analgesia in either naive or morphine-tolerant mice. In contrast, after intrathecal administration, cross-tolerance could be demonstrated between DPDPE or DPLPE and morphine, and no potentiation of morphine by DPDPE could be observed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3033213 TI - Use of beta-funaltrexamine to determine mu opioid receptor involvement in the analgesic activity of various opioid ligands. AB - Systemic administration of beta-funaltrexamine (beta-FNA) 24 hr before analgesic testing produced approximately a 10-fold parallel shift in the dose-response curves of the prototypic mu agonists morphine, I-methadone, fentanyl and etorphine in the mouse abdominal constriction test. In contrast, prior administration of beta-FNA produced no appreciable shift in the analgesic dose response curve of the selective kappa agonist, U-50, 488H. These results suggest that beta-FNA is selective for mu over kappa receptors under the conditions used in this study. The dose-response curves for ethylketazocine and proxorphan were affected only to a small extent by beta-FNA pretreatment, suggesting that these compounds have analgesic actions mediated primarily through nonmu, probably kappa receptors. The dose-response curves for cyclazocine, buprenorphine, butorphanol, nalorphine and nalbuphine were shifted markedly to the right and frequently not in a parallel fashion by the prior administration of beta-FNA. These results seem to indicate a major role for the mu receptor in the analgesic actions of these compounds. PMID- 3033215 TI - Developmental changes in muscarinic receptor-stimulated phosphoinositide metabolism in rat brain. AB - Muscarinic receptor-stimulated phosphoinositide hydrolysis was investigated in rat brain during ontogeny by measuring the accumulation of [3H]inositol phosphates ([3H]InsPs) in cerebral cortex slices at various ages. Experiments with carbachol and acetylcholine showed that [3H]InsPs accumulation was maximal in 7-day-old rats (1477 +/- 98% of basal) and lowest in adult (75 days) rats (428 +/- 24% of basal). No differences were found in the EC50 values for both cholinergic agonists. This effect appeared to be mediated by the M1-muscarinic receptor subtype as it was blocked by pirenzepine with Ki = 29.1 +/- 7.1 nM (adults) and 87.9 +/- 18.2 nM (7-day-old rats). Incorporation of [3H]inositol into phospholipid decreased from day 3 to adulthood; however, when data of [3H]InsPs release were corrected for the incorporation at a given age, the highest stimulation by cholinergic agonists was still observed in 7-day-old rats. Among the other neurotransmitters tested (norepinephrine, histamine and serotonin), all known to stimulate phosphoinositide metabolism, none had the same developmental profile of [3H]InsPs accumulation as cholinergic agonists. In contrast to carbachol- and acetylcholine-stimulated phosphoinositide hydrolysis, the density of muscarinic binding sites, measured by [3H]quinuclidinyl benzilate binding, increased from day 3 to day 75. Acetylcholinesterase activity also increased during development. The dissociation of receptor binding sites from receptor-stimulated phosphoinositide metabolism suggests the presence of a more effective receptor-effector coupling at specific times of neonatal development, particularly 1 week. Furthermore, the fact that maximal stimulation of phosphoinositide hydrolysis coincides with the period of brain growth spurt in the rats suggests that this system in the cerebral cortex might be involved in the processes of cell division and differentiation. PMID- 3033216 TI - Attenuation of the plasma prolactin response to restraint stress after acute and chronic administration of nicotine to rats. AB - We have previously shown that a single dose of nicotine elevates plasma adrenocorticotropin (ACTH) levels in rats and has a biphasic effect on plasma prolactin (PRL). The stimulatory effect of nicotine on these stress responsive hormones desensitizes after a single injection of nicotine. Continuous exposure to nicotine also induces tolerance to its locomotor depressive and hypothermic effects, which have been associated with an increase of central [3H]nicotine binding. Thus, the acute and chronic administration of nicotine might induce changes in central nicotinic cholinergic circuits that affect the ACTH and PRL responses to stress. In the present study, a single dose of nicotine (0.75-3.0 mg/kg b.wt.) significantly inhibited the elevation of plasma PRL due to restraint stress initiated 60 min afterward. Five injections of nicotine during 1 day produced a similar attenuation of the PRL response to restraint stress but neither of these paradigms affected ACTH. In contrast, intermittent delivery of nicotine for 7 days failed to affect the PRL response to restraint stress; however, after withholding nicotine for 14 hr, high dose nicotine attenuated the PRL response to stress, whereas low dose nicotine remained ineffective. On the other hand, administration of the same schedule of low dose nicotine did significantly diminish the expected release of PRL in response to a final injection of nicotine (0.5-2.0 mg/kg b.wt.) in unstressed animals. In summary, a single dose or 5 doses of nicotine in 1 day attenuated the PRL response to restraint stress, whereas, after chronic administration, this effect was lost.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3033218 TI - Studies on the agonistic activity of delta 9-11-tetrahydrocannabinol in mice, dogs and rhesus monkeys and its interactions with delta 9-tetrahydrocannabinol. AB - The present studies examine some of the pharmacological effects of delta-9 (11) tetrahydrocannabinol (delta 9-11-THC), an analog of delta-9-tetrahydrocannabinol (delta 9-THC). In tests with mice, delta 9-11-THC was similar to but less potent than delta 9-THC in producing hypothermia, analgesia, lethality and in reducing spontaneous activity. In dogs delta 9-THC but not delta 9-11-THC produced classical cannabimimetic signs including static ataxia, hyperreflexia, prancing and tail-tuck. delta 9-11-THC did produce central nervous system depression in 9 of the 15 dogs tested but the effects were not dose-related and appeared earlier and dissipated faster than the depressive effects induced by delta 9-THC. delta 9 THC but not delta 9-11-THC produced signs of ptosis, sedation and ataxia in rhesus monkeys. delta 9-THC also suppressed operant responding completely in four of four monkeys tested whereas in one monkey delta 9-11-THC did not do so up to doses as high as 5.0 mg/kg and was 8 to 100 times less potent in doing so in the other monkeys. When monkeys were pretreated with delta 9-11-THC the doses of delta 9-THC required to produce ptosis, sedation, ataxia and operant suppression were increased. However, when mice and dogs were pretreated with delta 9-11-THC the effects of delta 9-THC were not attenuated and usually were enhanced. The pharmacological profile of delta 9-11-THC is unusual in that it seems to have cannabimimetic activity in mice, noncannabimimetic-like effects in dogs and is perhaps devoid of cannabimimetic effects in rhesus monkeys. In addition, pretreatment with delta 9-11-THC attenuates the cannabimimetic effects of delta 9 THC in rhesus monkeys but not in mice or dogs. PMID- 3033217 TI - Effect of parathyroid hormone fragments on calcium transport in toad bladder. AB - Parathyroid hormone (PTH) has a variety of biologic effects which are both dependent and independent on activation of adenylate cyclase. We studied the effects of intact PTH and PTH fragments on water flow and Ca transport in isolated toad bladder sacs. As reported previously by us, PTH (1-84) significantly stimulated basal water flow in isolated toad bladder sacs. Synthetic PTH 1-34, 44-68, 53-84 and 65-84 (1 microgram/ml) had no effect on basal water flow after a 60-min incubation period. Intact PTH (1-84) and synthetic 1-34 PTH significantly inhibited both arginine-vasopressin and cyclic AMP-stimulated water flow. Synthetic PTH 44-68, 53-84 and 65-84 (1 microgram m/ml) had no effect on arginine-vasopressin or cyclic AMP-stimulated water flow after a 60-min incubation. Intact PTH (1-84) and synthetic 1-34 PTH significantly stimulated 45Ca uptake without affecting 45Ca efflux. Synthetic PTH 44-68, 53-84 and 65-84 had no effect on either 45Ca uptake or 45Ca efflux. Although these results suggest that the intact hormone is required for the maximal effect of PTH on water flow, substantial activity resides in the amino terminal fragment of the hormone. No activity per se resides in the carboxy terminal portion of the hormone as regards water flow. Alterations in Ca transport appear to mediate the effect of PTH on water flow. These effects are independent of activation of adenylate cyclase because these hormones also inhibit cyclic AMP-stimulated water flow. PMID- 3033219 TI - Modulation of rat brain opioid receptors by cannabinoids. AB - The interaction of delta 9-tetrahydrocannabinol (delta 9-THC) and related cannabinoids with opioid receptors of neuronal membranes has been investigated. Treatment of membranes with delta 9-THC consistently decreased specific in vitro binding of [3H]dihydromorphine (mu opioid) in a dose-dependent fashion. Similar dose-dependent changes were elicited by cannabidiol and (+/-) hexahydrocannabinol. Equilibrium binding studies in which brain membranes were titrated with [3H]dihydromorphine in the presence of delta 9-THC demonstrated that the decrease in [3H]dihydromorphine binding is due to a reduction in the number of binding sites, with no significant alteration in receptor affinity. This result suggests that the interaction of delta 9-THC with opioid receptors is a noncompetitive one. Delta 9-THC also inhibited the binding of the delta opioid [3H]D-Pen2, D-Pen5-enkephalin and the opioid antagonist [3H]naloxone (Ki = 16 and 19 microM, respectively) but failed to inhibit the binding of the kappa opioid [3H]ethylketocyclazocine (after suppression of mu and delta receptor binding), the phencyclidine analog [3H]N-(1-[2-theinyl]cyclohexyl)piperidine, the dopamine antagonist [3H]spiroperidol or the muscarinic antagonist [3H]quinuclidinyl benzilate. Moreover, delta 9-THC inhibited the binding of [3H]etorphine (potent opioid agonist) to solubilized, partially purified opioid receptors with a Ki value similar to that observed for the membrane-bound receptors. This finding indicates that the allosteric modulation of the opioid receptor by delta 9-THC is the result of a direct interaction with the receptor protein or with a specific protein-lipid complex and not merely the result of a perturbation of the lipid bilayer of the membrane.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3033220 TI - Effects of oxytocin-related peptides on acute morphine tolerance: opposite actions by oxytocin and its receptor antagonists. AB - The hormonally and behaviorally active nonapeptide oxytocin (OXT), its behaviorally active N-terminal octapeptide desglycinamide9-OXT and Z-prolyl-D leucine, a synthetic analog of the C-terminal prolyl7-leucine8 sequence, inhibited the development both of a moderate and of a strong tolerance to morphine. N-alpha-Acetyl-(2-0-methyltyrosine)-OXT and (penicillamine1-2-0 methyltyrosine)- lysine8-vasopressin, both OXT receptor antagonists, facilitated the development of a moderate morphine tolerance. The i.c.v. injection of either antagonist prevented the effects of i.c.v. and s.c. OXT treatment on the development of tolerance. The effect of desglycinamide9-OXT, but not that of Z prolyl-D-leucine was also prevented by N-alpha-acetyl-(2-0-methyltyrosine)-OXT. It is concluded that OXT and desglycinamide9-OXT, but not Z-prolyl-D-leucine, attenuate morphine tolerance by affecting putative oxytocinergic binding sites in the mouse brain. The fact that i.c.v. injection of the receptor antagonist also blocked the effect of s.c. OXT treatment argues in favor of the possibility that a minor proportion of s.c. OXT (or behaviorally active fragments thereof) may reach central nervous system target sites. PMID- 3033221 TI - Presynaptic alpha-2 adrenoceptors play a major role in the effects of idazoxan on cortical noradrenaline release (as measured by in vivo dialysis) in the rat. AB - Transcortical dialysis in awake unrestrained rats has been used to evaluate the functional role of differently located alpha-2 adrenoceptors in mediating the action of the alpha-2 adrenoceptor antagonist idazoxan on cerebral noradrenaline release. Basal efflux of noradrenaline collected by a cortically implanted dialysis fiber was stable over a period of 4 days. Systemic injections of idazoxan (20 mg/kg i.p.) increased cortical noradrenaline efflux. This effect was potentiated by pretreatment with the noradrenaline uptake blocker desipramine (20 mg/kg i.p.). Local cortical infusion of (10(-4) M idazoxan which provides a theoretical extracellular administration of 4 to 48 microM) via the dialysis fiber, thus eliminating the potential contribution of somatodendritic alpha-2 adrenoceptors, also elevated cortical noradrenaline efflux. Desipramine (20 mg/kg i.p.) potentiated this effect. Four days after lesioning cortical cell bodies with ibotenic acid (20 min infusion of 10(-4) M ibotenic acid via the dialysis fiber), both systemic injections and local cortical infusions of idazoxan were still effective in increasing cortical noradrenaline efflux. Lesion of serotonergic afferents to the cerebral cortex (by i.c.v. injection of 5,7 dihydroxytryptamine) or of cortical cholinergic afferents (by bilateral electrocoagulation of the nucleus basalis magnocellularis) did not affect the ability of cortical idazoxan infusion to stimulate noradrenaline efflux. The results suggest that the effects of idazoxan on cortical noradrenaline release are mediated primarily by alpha-2 adrenoceptors on noradrenergic nerve terminals, rather than by those located postsynaptically, somatodendritically or on the terminals of other neuronal inputs to the cerebral cortex. PMID- 3033222 TI - gamma-Aminobutyric acid receptor-regulated 36Cl- flux in mouse cortical slices. AB - An improved neurochemical assay for gamma-aminobutyric acid (GABA) function has been developed using tracer-radioactive chloride efflux in mouse cortical slices. Careful maintenance of the brain slice viability resulted in a 3-fold stimulation of 36Cl- efflux rate by the GABA agonist muscimol (EC50 = 3 microM), comparable to electrophysiologic and other chloride flux preparations. The shape of the muscimol dose-response curve was shallow, suggestive of negative cooperativity or heterogeneous receptors, but tissue uptake of agonist, possible diffusion barriers and apparent functional desensitization complicated these results. The response to muscimol was inhibited by GABAA receptor antagonists such as RU5135 and was enhanced by barbiturates and benzodiazepines. As observed previously, barbiturates stimulated 36Cl- efflux rate on their own and potentiated the response (potency and maximal effect) to muscimol in a stereospecific and picrotoxin-sensitive manner. Benzodiazepine receptor ligands alone did not alter 36Cl- flux, but agonists such as flunitrazepam enhanced the response to muscimol, an effect sensitive to the antagonist Ro15-1788. The inverse agonist methyl 6,7 dimethoxy-4-ethyl-beta-carboline-3-carboxylate did not inhibit muscimol-activated 36Cl- flux. The anthelminthic-insecticide avermectin B1a stimulated 36Cl- flux by itself, and this response was apparently additive with that of muscimol. This brain slice chloride flux assay is therefore suitable for the assessment of activity including dose-response curves for GABAA agonists, antagonists and modulators including benzodiazepines. PMID- 3033223 TI - Potentiation of aminoglycoside-induced neuromuscular blockade by protons in vitro and in vivo. AB - pH-dependent effects of 100 microM streptomycin and various aminoglycosides were examined at frog (Rana pipiens pipiens) sciatic sartorii in vitro by using the intracellular microelectrode recording technique. pH-dependent effects of streptomycin were also examined on indirectly elicited (nerve-stimulated) and directly elicited sartorius muscle twitches in vitro. Furthermore, in vivo effects of systemic pH alterations on neomycin-induced mortality were examined in Sprague-Dawley rats. There was a direct correlation between aminoglycoside potency and the number of basic groups per drug molecule (r = 0.95). At pH 7.2 and 9.0, the effect of pH on aminoglycoside potency correlated inversely with the pKa of the aminoglycoside (r = -0.98). At 0.7 and 2.2 pH units below 7.2, however, aminoglycoside-induced inhibitions of quantal content, end-plate potential amplitude and the indirectly elicited muscle twitch were potentiated by a pH-dependent mechanism that was independent of the pKa of the aminoglycoside. At these pH values, qualitatively similar drug effects were not observed on miniature end-plate potential amplitude and frequency or the directly elicited muscle twitch. Potentiation of aminoglycoside action was observed on mortality of rats, however, when the pH was 0.1 pH unit below 7.4. Thus, potentiation of aminoglycoside-induced neuromuscular blockade by protons in vitro and in vivo was demonstrated and appears to involve the pH-dependent activity of a prejunctional membrane component that regulates voltage-dependent, Ca++-mediated transmitter release. PMID- 3033225 TI - Dopaminergic and benzodiazepine receptors studied in vivo by PET. AB - Using Positron Emission Tomography (PET) and specific radioligands, dopaminergic D2 (DA-D2 receptors) and benzodiazepine receptors (BZ-receptors) were studied in living animals during normal and pathological conditions. In vivo characterization of both receptors was performed using two highly specific antagonists namely: 11C-Ro 15 1788 for BZ-receptors and 76 Br-Bromospiperone for DA-D2 receptors. Changes in 11c-Ro 15 1788 specific binding to BZ-receptors were observed during convulsive seizures. After MPTP treatment, a decrease in the 76 Br-Bromospiperone striatal specific binding was observed, correlated with the establishment of a Parkinson-like syndrome. PMID- 3033224 TI - Muscarinic release of inositol trisphosphate without mobilization of calcium in bovine adrenal chromaffin cells. AB - Chromaffin cells of bovine adrenal medulla release catecholamines in response to activation of nicotinic ACh receptors which open voltage-sensitive calcium channels. Catecholamine secretion by exocytosis requires an increase in cytosolic free calcium. The cells also possess muscarinic ACh receptors but muscarinic agents do not provoke catecholamine release. Quin-2 studies show that they do not increase cytosolic free Ca2+ concentration, but unlike the nicotinic agents, they cause phosphoinositide hydrolysis. Muscarinic stimulation leads to rapid loss of labelled phosphatidylinositol 4-phosphate and of phosphatidylinositol 4,5 bisphosphate. At the same time there is release of inositol trisphosphate, inositol bisphosphate and inositol phosphate. In a number of other cells inositol trisphosphate may act as a second messenger releasing Ca2+ from storage sites in the endoplasmic reticulum but this is not its function in bovine chromaffin cells. PMID- 3033226 TI - Protein phosphorylation and synaptic transmission: receptor mediated modulation of protein kinase C in a rat brain fraction enriched in synaptosomes. AB - Aspects of protein phosphorylation related to events occurring during synaptic transmission were briefly reviewed. High resolution two-dimensional electrophoresis was used to study protein phosphorylation catalysed by protein kinase C in a fraction from rat brain enriched in synaptosomes. Incubation of 32P labelled synaptosomes with 4 beta-phorbol 12 beta-myristate 13 alpha-acetate resulted in an increase in the phosphorylation of a 45 K polypeptide (generally known as B-50) and an 82 K polypeptide; other major phosphoproteins in the preparation were unaffected by this treatment. It appears therefore that the 45 K and 82 K polypeptides are the only significant substrates for protein kinase C in synaptosomes. Depolarisation of labelled synaptosomes by high K+ increased the phosphorylation of the 82 K polypeptide, synapsin I and several unknown phosphoproteins. Incubation of labelled synaptosomes with the cholinergic agonist carbachol resulted in a modest, but statistically significant, increase in the phosphorylation of the 45 K (B-50) and 82 K polypeptides. This effect was blocked by atropine. The results are discussed in relation to a possible role for the B 50 phosphoprotein in regulating the resynthesis of polyphosphoinositides following cholinergic stimulation. PMID- 3033227 TI - Immunoreactive forms of ACTH released by the adenohypophysis of the rat during the perinatal period in vivo and in vitro studies. AB - Immunoreactive ACTH levels were determined in whole plasma, eluate fractions of plasma and perfusates of anterior lobe extracts subjected to chromatography on Sephadex G-50 fine. In the fetal plasma, ACTH levels were higher on day 19 than on days 17, 18, 20 and 21. After birth, ACTH concentrations dropped to reach the lowest values in one week old newborns; thereafter they increased until weaning on day 21. They were then similar to those of non pregnant adult females. Three peaks of immunoreactive ACTH were present in all the chromatograms; the first one eluted near the void volume ("big" ACTH, PM approximately 44,000), the third one coeluted with human ACTH (1-39) ("little" ACTH, PM approximately 4,500) and the second one eluted midway between the 2 previous peaks ("intermediate" ACTH, PM approximately 13,000). During the last days of pregnancy, the proportion of the "little" form of ACTH in the fetal plasma showed a gradual increase whereas that of the "big" one decreased. On days 17, 19 and 21 of gestation the anterior lobes of fetal pituitary glands released in vitro these 3 forms of immunoreactive ACTH in the same proportions as those observed in the anterior lobes. In contrast, the proportions of the circulating forms of ACTH were quite different; the "little" one gradually increased and the "big" one decreased as gestation progressed. In vitro controlled tryptic digestion of the isolated "big" form led to the appearance of "intermediate" and "little" forms suggesting some transformations in the circulation of the ACTH forms released by the fetal hypophysis in vivo. PMID- 3033228 TI - [Effect of marathons on neuromuscular excitability; comparison of electromyographic and biochemical findings]. AB - The aim of this paper is to study the neuromuscular excitability of a group of marathon runners and to see how it can be modified right after the marathon. Spontaneously appearing multiplets in EMG signal under ischaemia served as a test of the neuromuscular hyperexcitability. The percentage of positive tests is much higher than in the control population; as a rule, the effect of the marathon is to diminish the neuromuscular hyperexcitability. The concentrations of Ca, Mg and P in the plasma as well as the globular concentration of Mg have been determined several times, both before and after the marathon; no correlation has been found with the neuromuscular hyperexcitability. PMID- 3033229 TI - [Effects of parathyroid hormone on the cardiovascular system]. AB - It has been well accepted that the bone and kidney are the principal organs of parathyroid hormone (PTH) actions, but there has been little work on the cardiovascular system. We evaluated the effect of PTH on the cardiovascular system of rats. In thiobutabarbital anesthetized rats, synthetic bovine parathyroid hormone, containing the amino acid (b-PTH 1-34) in dose of 0.1-10 micrograms/kg iv, caused dose related decrease in mean arterial blood pressure (MAP). On the other hand, there were significantly increase in heart rate (HR) and left ventricular contractile force. With the doses of 10 micrograms/kg, PTH decreased the MAP from 104.3 to 55.5 mmHg, left ventricular pressure (LVP) 122.1 to 96.4 mmHg, left ventricular end diastolic pressure (LVEDP) from 6.70 to 6.37 mmHg and LV dp/dt max 5,684 to 4,736 mmHg/sec. The HR, LV dp/dt/p and Vmax increase from 399.7 to 410.0 bpm, 95.5 to 108.4/sec, 98.2 to 107.4/sec, respectively. The propranolol, phentolamine, atropine and promethazine did not affect these actions of PTH. On the basis of these findings, we conclude that PTH has the directory vasodepressive action and the effect of augmentation of the left ventricular contractile force. PMID- 3033230 TI - Evaluation of implanted durapatite particles in fresh extraction sockets to maintain the alveolar ridge in beagle dogs. PMID- 3033231 TI - Fabrication of a sectional surgical stent for hydroxyapatite augmentation for the edentulous residual ridge. PMID- 3033232 TI - Inhibition of ovulation in PMSG/hCG-treated immature rats by rotenone, a specific inhibitor of mitochondrial oxidation. AB - Immature Wistar rats were induced to ovulate by treatment with PMSG and hCG. Control animals ovulated 43.5 +/- 0.36 ova/rat. Intraperitoneal injection of rotenone doses of 0.125, 0.25 and 0.50 mg/kg reduced the ovulation rate to 24.0 +/- 3.08, 8.0 +/- 0.88 and 1.5 +/- 0.44 ova/rat, respectively. The rotenone significantly reduced ovarian cytochrome oxidase activity and progesterone production, but not production of oestradiol or testosterone. Thyroxine treatment at a dose of 5 mg/kg s.c. reversed the rotenone inhibition of ovulation. The results suggest that an increase in mitochondrial respiration is an essential feature of the ovulation process in mammals. PMID- 3033234 TI - Fetal viral myocarditis and congenital complete heart block in a pregnancy complicated by systemic lupus erythematosus. A case report. AB - Congenital complete heart block has been associated with collagen vascular disorders. Many etiologies have been proposed for the myocarditis that develops and results in the complete heart block. Transplacental antibody transfer- specifically, antibodies to tissue-soluble ribonuclear protein antigen-A--has been implicated as the etiology of the myocarditis. This case supports the association of this antibody while suggesting that viral infection may serve as an initiator of the autoimmune destruction. Also, fetal viral infection alone must be considered a possible cause of congenital complete heart block. PMID- 3033233 TI - Effects of a luteolytic dose of oestradiol benzoate on uterine oxytocin receptor concentrations, phosphoinositide turnover and prostaglandin F-2 alpha secretion in sheep. AB - Administration of oestradiol-17 beta benzoate on Days 9 and 10 of the oestrous cycle resulted in episodic secretion of PGF-2 alpha (as indicated by elevated circulating concentrations of 13,14-dihydro-15-ketoprostaglandin F-2 alpha) and a decline in circulating progesterone. Release of PGF-2 alpha began 35 +/- 3 h after first injection of oestrogen and progesterone concentrations declined from 42 +/- 3 h. Secretion of oxytocin, which was first observed 26 +/- 3 h after oestrogen treatment, preceded secretion of PGF-2 alpha; 69% of pulses of oxytocin coincided with episodes of PGF-2 alpha secretion. Uterine oxytocin receptor concentrations were raised in ewes treated with oestrogen, increases occurring in caruncular endometrium and myometrium by 12 h after treatment and in intercaruncular endometrium by 24 h. Raised receptor concentrations were followed at 24 h by increases in the incorporation of [3H]inositol into phosphatidylinositol and in the hydrolysis of labelled tissue phosphoinositides in response to oxytocin in slices of caruncular endometrium incubated in vitro. The following sequence of events is therefore suggested to occur at oestrogen induced luteolysis: induction of the oxytocin receptor; increased turnover of phosphoinositides; onset of episodic secretion of PGF-2 alpha; and functional luteolysis. PMID- 3033235 TI - Coxsackievirus B 1 induced murine polymyositis: acute infection with active virus is required for myositis. AB - Although the etiology of human polymyositis (PM) remains obscure, group B coxsackieviruses (CVB) have been implicated in disease pathogenesis and a particular strain of type 1 CVB (CVB 1) has been shown to cause in mice an inflammatory myositis which is similar to human PM. After infection of neonatal Swiss mice with active CVB 1, virus replicated to high titers in muscle and produced acute myonecrosis. Viral titers peaked in muscle at Day 7 and virus was undetectable after Day 14. Myocytes regenerated, but a chronic inflammatory myositis, involving principally proximal muscle groups of the hind limbs, was observed by Day 14 and persisted through at least Day 70. Injection of inactivated virus resulted in no clinical or histological disease. Anti-CVB 1 antibody was demonstrated in mice injected with inactivated or active virus pools. This report establishes the need for active virus in the pathogenesis of CVB 1 induced murine PM. PMID- 3033236 TI - Giant cell arteritis and peripheral neuropathy: a report of 2 cases and review of the literature. AB - Giant cell arteritis (GCA) is a systemic vasculitis primarily affecting large and medium sized vessels. While the disease may present with blindness or other signs of extracranial vasculitis, symptoms referable to the peripheral nervous system are uncommon. We describe 2 patients with biopsy proven GCA who simultaneously developed peripheral neurologic lesions. The first developed a mononeuritis multiplex superimposed on a diffuse, primarily sensory and distal polyneuropathy; the second, a symmetric, primarily motor and distal polyneuropathy. We review the published experience of GCA with peripheral nerve involvement and discuss a possible pathophysiologic basis for its occurrence. PMID- 3033237 TI - Tetracyclines inhibit human synovial collagenase in vivo and in vitro. AB - To determine if tetracyclines can inhibit human synovial collagenase from rheumatoid tissue, paired synovial tissue (or synovial fluid) was collected from 7 patients before and after oral administration of minocycline (100 mg BID) for 10 days. With each patient serving as his own control, the postminocycline collagenase activities fell an average of 67% from pretreatment values. Qualitative SDS-PAGE revealed decreased loss of alpha collagen components and reduced formation of alpha A digestion fragments. Addition of minocycline or a chemically modified tetracycline to synovial culture media in vitro profoundly inhibited collagenase activity. Further study of this action of tetracyclines could serve as a probe of the role of collagenase in rheumatoid arthritis and lead to development of agents capable of modifying the tissue destructive actions of collagenase. PMID- 3033238 TI - Effects of recombinant human interferons on rheumatoid arthritis B lymphocytes activated by Epstein-Barr virus. AB - We evaluated the effects of all 3 classes of recombinant human interferon (IFN) on Epstein-Barr virus (EBV) infection of purified B lymphocytes from patients with rheumatoid arthritis (RA). After EBV infection, RA B cells secreted more IgM and significantly more IgM rheumatoid factor (RF) than normals. Spontaneous (no EBV) proliferation, IgM, and IgM RF were also higher in RA. All 3 types of IFN inhibited dose dependently EBV induced B cell activation. In RA, however, higher doses of each class of IFN were necessary to obtain 50% inhibition. IFN gamma was most potent in normals and RA. Four IgM RF production IFN gamma was significantly more potent than IFN alpha and IFN beta in reducing the spontaneous activation of RA B cells, and a similar trend was seen in B cell proliferation. These findings are discussed in the context of ongoing clinical trials with IFN gamma in RA. PMID- 3033239 TI - Reversal of depressed neutrophil superoxide production in Felty's syndrome after gold therapy. AB - Six patients with Felty's syndrome were reevaluated after a course of chrysotherapy. In 5 patients, white cells and absolute neutrophil counts had returned to normal. In 4 of these, there was significant concomitant improvement in neutrophil function as assessed by the ability of cells to generate superoxide radicals. PMID- 3033240 TI - Effect of low dose methotrexate on neutrophil chemotaxis induced by leukotriene B4 and complement C5a. AB - When mediators of inflammation such as complement component C5a or leukotriene B4 are introduced into an air pouch created in mice, these mediators induce the migration of neutrophils into the air pouch. Pretreatment of mice with low doses of methotrexate inhibits leukotriene B4 or C5a induced neutrophil migration into the air pouch. Inhibition of neutrophil chemotaxis by methotrexate may, at least in part, account for the rapid onset of antiinflammatory activity that was observed in clinical trials with methotrexate in rheumatoid arthritis. PMID- 3033241 TI - Dihydropyridazinone cardiotonics: synthesis and inotropic activity of 5'-(1,4,5,6 tetrahydro-6-oxo-3-pyridazinyl)spiro[cycloalkane- 1,3'-[3H]indol]-2'(1'H)-ones. AB - In the 1,3-dihydro-5-(1,4,5,6-tetrahydro-6-oxo-3-pyridazinyl)-2H-indol-2-one series of cardiotonics, we found that a spirocycloalkyl ring may be annealed to the 3-position of the indolone moiety while retaining inotropic activity. An inverse relationship was found between spirocyloalkyl ring size and inotropic potency. ED50 values of the spirocyclopropane 10, spirocyclobutane 12, and spirocyclopentane 13 were 2.7, 35, and 133 micrograms/kg, respectively, following iv administration to pentobarbital-anesthetized dogs. The most potent compound prepared was 11 (5'-(1,4,5,6-tetrahydro-4-methyl-6-oxo-3 pyridazinyl)spiro[cyclopropane- 1,3'-[3H]indol]-2'(1'H)-one), the 4-methyl analogue of 10. This compound had an iv ED50 of 1.5 microgram/kg. Oral activity was evaluated by administering 50 micrograms/kg of 10 to conscious, chronically instrumented dogs. A 39% increase in LV dP/dt60 was observed, and an inotropic effect was demonstrable in excess of 7 h. Thus, the spirocyclic dihydropyridazinone inotropes are potent, long-acting, orally effective cardiotonics. Compound 11 was a potent inhibitor (IC50 = 13 nM) of cAMP phosphodiesterase derived from canine cardiac sarcoplasmic reticulum (SR-PDE). Importantly, -log IC50 values for inhibition of SR-PDE for this entire series of compounds were highly correlated (r = 0.949, p less than 0.02) with their inotropic -log ED50 values, supporting the hypothesis that inhibition of this enzyme contributes to the mechanism of action of the spirocyclic dihydropyridazinones. PMID- 3033242 TI - Estrogenic affinity labels: synthesis, irreversible receptor binding, and bioactivity of aziridine-substituted hexestrol derivatives. AB - To develop an affinity label for the estrogen receptor that would be an estrogen agonist, rather than antagonist, we prepared several aziridine derivatives of the potent nonsteroidal estrogen hexestrol [3R,4S)-3,4-bis(4-hydroxyphenyl)hexane) bearing an aziridine function on the side chain. Three functional groups link the hexestrol ligand and the aziridine: a carbonyl group (ketone or ester), a thioether, or a methylene chain. The apparent competitive binding affinity of these derivatives for the estrogen receptor ranges from 1.8% to 25% that of estradiol, and most of them bind in a time-dependent, irreversible manner with the receptor, although the rate and efficiency of this binding vary widely, often with relatively small changes in structure. This is consistent with the irreversible attachment requiring a precise alignment of activating and reacting residues in the binding site of the receptor. The estrogenic and antiestrogenic activity of these aziridine derivatives was investigated in MCF-7 human breast cancer cells. Most of the compounds are agonists, with one being an antagonist. The derivative (6R,7S)-1-N-aziridinyl-6,7-bis(4-hydroxyphenyl)-5-nonanone (keto nonestrol aziridine 3) appears to have the most ideal behavior of the estrogenic affinity labeling agents prepared: It is an agonist, and it binds to receptor irreversibly, efficiently, and quite rapidly. PMID- 3033243 TI - Potential anti-AIDS drugs. 2',3'-Dideoxycytidine analogues. AB - 5-Substituted 2',3'-dideoxycytidine analogues have been synthesized and evaluated in vitro for their capabilities to protect T4+ lymphocytes from the cytopathic effects of the HTLV-III/LAV (HIV) virus, the causative agent of acquired immunodeficiency syndrome (AIDS). These analogues were designed to be more lipophilic than 2',3'-dideoxycytidine (ddC) in order to enhance central nervous system penetration. Earlier reports had shown that ddC is a potent protective agent. When ddC is substituted at the 5-position with either methyl or bromo substituents, activity is completely abolished. However, when the substitution is fluoro (5-F-ddC), both activity and potency are retained. 2',3'-Dideoxy-5 azacytidine is also protective but more toxic than ddC or 5-F-ddC. In a different approach, an attempt was made to utilize ddCMP, ddTMP, and ddAMP as preformed nucleotides in order to circumvent the generally low level of phosphorylation achieved with dideoxynucleosides which function as relatively poor substrates for the cellular kinases. Only ddAMP is as active as its nucleoside precursor. Because ddAMP is not more active than ddA at low concentrations, it is possible that the active agent is ddA which is generated from ddAMP prior to cell entry. PMID- 3033245 TI - Leukotriene receptor antagonists. 2. The [[(tetrazol-5 ylaryl)oxy]methyl]acetophenone derivatives. AB - A series of [[(tetrazol-5-ylaryl)oxy]methyl]acetophenones was synthesized and evaluated as antagonists of leukotriene D4 induced contractions of guinea pig ileum. Substitutions at the 3-position of the acetophenone with ethyl (66), propyl (68), butyl (83), and isobutyl (84) gave -log IC50 values of 7.9, 8.0, 7.8, and 7.7, respectively. Equally potent compounds were obtained when the tetrazol-5-yl group was connected to the second benzene ring in the para position with a chemical bond (67), methylene (68), or ethylene (71). For retention of high antagonist activity, the acetophenone should be substituted in the 2 position by a hydroxyl group and the tetrazole ring should have an acidic hydrogen atom. 1-[2-Hydroxy-3-propyl-4-[[4-(1H-tetrazol-5-ylmethy) phenoxy]methyl]phenyl]ethanone (68, LY1632443) has undergone extensive pharmacologic evaluation for its potential as an antiasthma agent. PMID- 3033244 TI - 1-(2-Deoxy-2-fluoro-beta-D-arabinofuranosyl)-5-ethyluracil. A highly selective antiherpes simplex agent. AB - The protected nucleoside 1-(2-deoxy-2-fluoro-3,5-di-O-benzoyl-beta-D arabinofuranosyl)-5-ethylura cil (10) was prepared by condensation of 3,5 dibenzoyl-2-deoxy-2-fluoro-alpha-D-arabinofuranosyl bromide (9) with 2,4-bis-O (trimethylsilyl)-5-ethyluracil (8). The ratio in this coupling reaction has been raised to 17:1 in favor of the desired beta-anomer. Deprotection by aminolysis gave 1-(2-deoxy-2-fluoro-beta-D-arabinofuranosyl)-5-ethyluracil (FEAU, 1) in 67% isolated yield from the bromo sugar 9. In vitro data show that FEAU has activity against herpes simplex virus types 1 and 2 comparable to that of 1-(2-deoxy-2 fluoro-beta-D-arabinofuranosyl)-5-methyluracil (FMAU, 2), 1-(2-deoxy-2-fluoro beta-D-arabinofuranosyl)-5-iodouracil (FIAU, 3), and acyclovir (ACV, 12). The cellular toxicity of FEAU was found to be much lower than that of the other nucleoside analogues. Biochemical experiments indicate that FEAU has similar affinity toward thymidine kinases encoded by HSV 1 and 2 and a much lower affinity for cellular thymidine kinase than thymidine. The in vivo antiviral effects of FEAU, FMAU, FIAU, and ACV were evaluated against herpes infection in a systemic mouse encephalitis model and a cutaneous guinea pig model. While FEAU showed activity comparable to that of ACV in the systemic infection model, it was superior in the cutaneous herpes infection model. PMID- 3033246 TI - Opioid agonist and antagonist activities of monofunctional nitrogen mustard analogues of beta-chlornaltrexamine. AB - In an effort to compare the role of a monofunctional nitrogen mustard with that of its bifunctional counterpart (i.e., beta-CNA, 1b) in modulating nonequilibrium activity of opioid receptors, we have synthesized and tested N-(2-chloroethyl)-N methylamino analogues 2a and 2b. Compound 2b and beta-CNA (1b) possessed qualitatively similar pharmacologic profiles on the guinea pig ileum (GPI) and mouse vas deferens (MVD) preparations. Moreover, the corresponding epimer 2a behaved somewhat like that reported for alpha-CNA (1a) in that it possessed irreversible agonist activity in the GPI. The similar pharmacologic profiles of the monofunctional and bifunctional nitrogen mustards suggest that possible cross linking of receptor nucleophiles by the latter is not critical for activity. In addition, the results are consistent with the idea that the rank-order nonequilibrium activity of 2b at different opioid receptor types is related to its relative affinity at those sites rather than to the alkylation step. PMID- 3033247 TI - Synthesis and biological activity of unsaturated carboacyclic purine nucleoside analogues. AB - Two new carboacyclic nucleoside analogues, 9-[4-hydroxy-3-(hydroxymethyl)-2 butenyl]adenine (6) and 9-[4-hydroxy-3-(hydroxymethyl)-2-butenyl]guanine (5), modeled on the unsaturated carbocyclic nucleoside analogue neplanocin A (2), have been synthesized and tested for antiviral activity against HSV-2 and, in the case of 6, for activity against influenza and in vitro inhibition of S adenosylhomocysteine hydrolase. The synthesis was accomplished through the coupling of either adenine or the guanine precursor 2-amino-6-chloropurine (15) to the key intermediate 1-(benzyloxy)-2-[(benzyloxy)methyl]-4-chloro-2-butene (13). Debenzylation of the N-9 adenine adduct gave 6 directly, while the product of the debenzylation of the N-9 adduct of 15 when treated with sodium hydroxide gave the guanine analogue 5. The carboacyclic guanine analogue (5) exhibited significant antiviral activity against HSV-2 (VR = 1.5, MIC50 = 65.6 micrograms/mL), a level of activity that is superior to that of ara-A but inferior to that of acyclovir. The adenine analogue 6 was active against HSV-2 only at a very high dose; it was devoid of antiviral activity against influenza type A2, and it lacked inhibitory activity against S-adenosylhomocysteine hydrolase. PMID- 3033249 TI - An abdominal mass: the congenital mesoblastic nephroma (CMN). The pathologic basis of its management. PMID- 3033250 TI - Effect of isoproterenol on myocardial mechanical function and cyclic AMP content in the fetal rabbit. AB - The effect of isoproterenol on mechanical function was studied in the isolated arterially perfused heart of the fetal (21st and 28th day of gestation) and newborn rabbits. The inotropic effect of isoproterenol in the fetus was less than in the newborn. In contrast, myocardial cyclic AMP levels after isoproterenol infusion in the fetus were greater than in the newborn. The inotropic effects of dibutyryl cyclic AMP and of calcium in the fetus were less than in the newborn. These data suggest that the process from beta-receptor to cyclic AMP in the fetus was equally or even more responsive to isoproterenol than in the newborn. The diminished inotropic effect of isoproterenol in the fetus may be due, at least in part, to the decreased inotropic effect of calcium. PMID- 3033248 TI - Hexahydro-1H-1-pyrindines from acid rearrangement of 9-alkylidene-5-(m methoxyphenyl)-2-methylmorphans. A new structural type of narcotic antagonists. AB - 9-Methylene- and 9-ethylidene-5-(m-methoxyphenyl)-2-methylmorphans (1, 2) and refluxing 48% HBr have given rearrangement products 3 and 4, respectively. The structure of 4 [4a-ethyl-2,4a,5,6,7,7a-hexahydro-4-(3-hydroxyphenyl)-1-methyl-1H 1- pyrindine] was determined by X-ray crystallography and that of 3 [1,4a dimethyl-2,4a,5,6,7,7a-hexahydro-4-(3-hydroxyphenyl)-1-methyl-1H- pyrindine] follows from analogy and NMR data. Compounds 3 and 4 are opioid antagonists of about the potency of nalorphine in the tail-flick vs. morphine assay and precipitate a complete abstinence syndrome in morphine-dependent monkeys. Both are nearly devoid of antinociceptive activity and they have about 0.025 times the affinity of nalorphine for the mu opioid receptor. PMID- 3033252 TI - Calibration of mitochondrial DNA evolution in geese. AB - Mitochondrial DNA was purified from five American species of geese representing the genera Anser and Branta, which have fossil records. The results of electrophoretic comparisons of about 75 fragments per individual produced by 14 restriction enzymes imply that the mean extent of sequence divergence between species of Anser and Branta is about 9%. Fossil evidence suggests that these two groups of geese had a common ancestor 4-5 million years ago. Thus, the mean rate of sequence divergence in goose mitochondrial DNA is not far from 2% per million years, the value in mammals. PMID- 3033254 TI - Asbestos-induced changes in rat lung parenchyma. AB - Fischer 344 rats have been exposed to UICC crocidolite by whole-body inhalation procedures for periods of 1 d to 12 mo. Material was obtained from the same location in the left lung, and the numbers of cells in the parenchyma were identified and determined by transmission electron microscopy. An immediate increase (1 d of exposure) was evident in the number of type II cells, suggesting a direct action of the dust on these cells. The number of interstitial and alveolar macrophages showed a significant increase after 3 mo of exposure. The number of alveolar macrophages containing dust particles after a 1-d exposure was 49%, and the corresponding value after 12 mo of exposure was 92%. The longer periods of exposure were associated with an increase in the number of particles per macrophage. Polymorphs appeared in the interstitium at airway bifurcations, prior to their appearance in the alveolar space. These bifurcations were also the initial sites where evidence of cell damage and collagen deposition was seen. In this experiment crocidolite appears to be weakly fibrogenic, and other factors may be needed to produce the marked lesions seen in human asbestosis. PMID- 3033251 TI - Reduction in the activity of the stimulatory guanine nucleotide-binding protein in the myocardium of spontaneously hypertensive rats. AB - The beta-adrenergic receptor-adenylate cyclase system of the cardiac membranes in spontaneously hypertensive rats (SHR) 14 weeks old was studied. The maximal activity of the catalytic unit of adenylate cyclase stimulated by purified stimulatory guanine nucleotide-binding protein (Ns) or forskolin was higher in SHR than in control Wistar-Kyoto (WKY) rats. However, adenylate cyclase activity stimulated by isoproterenol and GTP was the same between SHR and WKY rats. Although there was no difference in the amount of Ns which was measured by cholera toxin-catalyzed ADP-ribosylation, the functional activity of Ns in cholate-extracted membranes from SHR was significantly lower than that from WKY rats. There were no strain differences in the number and affinity of beta adrenergic receptors; the function and amount of the inhibitory guanine nucleotide-binding protein (Ni), and the amount of beta gamma-subunits of Ns and Ni. These findings showed that there is an abnormal signal transduction in this system in SHR due to a reduction in the functional activity of alpha-subunits of Ns. PMID- 3033255 TI - Binding of nerve growth factor to its receptor. PMID- 3033253 TI - Survival of patients with unresectable non-small cell lung cancer following 6,000 rad megavoltage radiotherapy. AB - One hundred and fifty-seven patients with histologically proven, locally advanced, unresectable or inoperable non-small cell lung cancer were treated in a uniform fashion with external-beam megavoltage radiotherapy. Patients received a continuous course of 6,000 rad in 71/2 weeks (four fractions of 200 rad per fraction each week). The comparatively high, uncorrected one-year survival rate of 48.7 percent, and uncorrected two-year survival rate of 25.7 percent, would be expected following uniform treatment with a relatively high external-beam tumor dose. The five-year uncorrected actuarial survival rate of 2.7 percent demonstrates that although the median survival can be increased by the use of higher radiation doses and better delivery technique, the ultimate cure rate and prognosis for inoperable and unresectable non-small cell lung cancer remains poor with current equipment and methods of radiotherapy. PMID- 3033256 TI - Neuritic growth from a new subline of PC12 pheochromocytoma cells: cyclic AMP mimics the action of nerve growth factor. AB - We have identified a new subline of PC12 pheochromocytoma cells (PC12D cells) in which neurites are extended within 24 hr in response to cAMP-enhancing reagents as well as in response to nerve growth factor (NGF), but not in response to epidermal growth factor or phorbol diester. Anti-NGF antiserum did not affect forskolin (FRK)-induced neuritic recruitment. FRK-induced neurites exhibited growth cones and contained secretion granules and many parallel arrays of microtubules as was the case with NGF-induced neurites. FRK, but not NGF, increased the levels of intracellular cAMP and activated adenylate cyclase in the membrane fraction. Both NGF and FRK enhanced the activities of tyrosine hydroxylase (TH), acetylcholinesterase (AchE), and ornithine decarboxylase (ODC), but not the levels of neuron-specific enolase. Enhanced levels of intracellular cAMP mimicked the effects of NGF on neuritic growth, TH, AchE, and ODC activities in PC12D cells, even though NGF does not act through elevation of levels of cAMP. PMID- 3033257 TI - Ganglioside-induced alterations in hippocampal cholinergic enzymes and Na,K ATPase after fimbria-fornix transection. AB - Previous biochemical findings suggest that exogenous gangliosides enhance cholinergic sprouting in the hippocampus after partial lesions of the septohippocampal pathway. To assess whether GM1 ganglioside accelerates the onset of this sprouting after complete lesions, we measured cholinergic enzymes and Na,K-ATPase activity in the hippocampus of rats with unilateral fimbria-fornix transection. At 14 and 18 days postlesion, histochemical staining showed that acetylcholinesterase (AChE) was almost completely eliminated in the hippocampus ipsilateral to the transection in untreated and GM1-treated rats. Biochemical assays confirmed that GM1 treatments did not increase AChE activity in the denervated hippocampus. Rather, there were significant reductions of AChE and choline acetyltransferase activities in the ipsilateral hippocampus relative to the contralateral value (P less than .001); and the reductions were greater in GM1-treated rats than in untreated controls (P less than .001). Na,K-ATPase activity in the ipsilateral hippocampus increased by 10.1% in GM1-treated rats, whereas it decreased by 21.7% in untreated controls (P less than .05). Since Na, K-ATPase is enriched in synaptic membranes, the increased activity of this enzyme may indicate that GM1 treatments stabilize surviving synaptic membranes and/or accelerate the onset of sprouting in the denervated hippocampus. The reductions in cholinergic enzymes, however, imply that the sprouting pathway must be noncholinergic. PMID- 3033258 TI - Effects of arginine vasopressin on protein phosphorylation in rat hippocampal synaptic membranes. AB - Our laboratory has reported previously the characteristics of specific AVP binding to rat hippocampal synaptic membranes (SPM) in the presence of Ni2+ [Costantini MG, Pearlmutter AF: J Biol Chem 259: 11739-11745, 1984]. We extended our investigation to determine the effects of Ni2+, (AVP), and AVP analogs on SPM protein phosphorylation. Ni2+ (5 mM) caused a dramatic reduction in phosphorylation of most SPM phosphoproteins. The most prominent protein which is phosphorylated in SPM has a molecular weight of 48 kilodaltons (KDa) and has been named B50 or F1; this protein shows altered phosphorylation in vitro in response to long-term potentiation in vivo as well as changes induced by exposure of SPM to ACTH (1-24), dopamine, and somatostatin. AVP and related peptides reduced phosphorylation of this pre-synaptic phosphoprotein in the following order of potency: AVP = oxytocin greater than DG-AVP greater than dDAVP greater than d(CH2)5Tyr(Me)AVP = [pGlu4,Cyt6]AVP-(4-9). Except for the pressor antagonist d(CH2)5Tyr(Me)AVP, this corresponds to their relative efficacy in displacing 3H AVP from high-affinity specific binding sites on rat hippocampal synaptic membranes. Ni2+ did not alter the degree of inhibition caused by the peptides. When SPM were treated with AVP after the attainment of maximum 32P incorporation, AVP inhibited dephosphorylation over a 30-min period. Our results show that AVP can alter both phosphorylation and dephosphorylation of hippocampal SPM phosphoproteins in vitro; the direction of these effects depends upon experimental conditions. Since B50/F1 is known to be a substrate for protein kinase C, AVP may act by inhibition of protein kinase C activity, either directly or indirectly. PMID- 3033259 TI - The effect of oropharyngeal decontamination using topical nonabsorbable antibiotics on the incidence of nosocomial respiratory tract infections in multiple trauma patients. AB - The incidence of respiratory tract infections was determined in 59 multiple trauma patients requiring prolonged intensive care (greater than 5 days) and receiving no antibiotic prophylaxis. Early pneumonia (less than 48 hr) with S. aureus, S. pneumoniae, and/or H. influenzae was found in 44% of patients. Secondary colonization of the oropharynx and respiratory tract with ICU associated Gram-negative bacilli followed by pneumonia occurred in 12 patients (20%). The overall incidence of respiratory tract infections was 59%. In a prospective open trial three prophylactic antibiotic regimens were compared: 17 patients were treated with intestinal decontamination using nonabsorbable antibiotics (polymyxin E 400 mg, tobramycin 320 mg, amphotericin B 2,000 mg/day). No difference in infection rate was found. Twenty-five patients were treated with intestinal and oropharyngeal decontamination using an ointment containing 2% of the same antibiotics. Secondary colonization and infection of the respiratory tract with Gram-negative bacilli was significantly reduced (p less than 0.001). The incidence of early (Gram-positive) infections, however, was unchanged. Another group of 63 patients was treated with systemic antibiotic prophylaxis during the first days in combination with oropharyngeal and intestinal decontamination. The incidence of early pneumonia was significantly reduced (p less than 0.001). Five patients (8%) developed an infection. Superinfections were not observed. PMID- 3033260 TI - Relationships among plasma cortisol, adrenocorticotrophin, and severity of injury in recently injured patients. AB - When samples were taken within 2 hr of accidental injury, plasma cortisol levels increased with injury severity score (ISS) in patients with minor and moderate injuries (ISS, 1-12) but decreased at higher ISS. Radioimmunoassay of adrenocorticotrophin (ACTH) showed very variable plasma concentrations at all ISS ranges. Like cortisol, ACTH increased with ISS up to a score of about 13, but it thereafter plateaued, and in patients with severe injuries plasma cortisol fell in relation to ACTH as well as in absolute terms. This suggested that although ACTH secretion was generally far from maximal the relatively low cortisol concentrations in the most severely injured were at least partly due to a poor response of the adrenal cortex to ACTH. In patients who presented late (more than 2 hr after injury) plasma cortisol levels were more variable and more strongly related to ACTH than at shorter times after injury. The variability of cortisol also increased with age. Patients with severe head injuries had cortisol and ACTH concentrations similar to those without head injuries but with a similar ISS from injuries in other parts of the body. PMID- 3033261 TI - Influence of the Epstein-Barr virus nuclear antigen EBNA 2 on the growth phenotype of virus-transformed B cells. AB - Epstein-Barr virus (EBV) isolates show sequence divergence in the BamHI YH region of the genome which encodes the nuclear antigen EBNA 2, a protein thought to be involved in the initiation of virus-induced B-cell transformation; type A isolates (such as B95-8 EBV) encode a 82- to 87-kilodalton EBNA 2A protein, whereas type B isolates (such as AG876 EBV) encode an antigenically distinct 75 kilodalton EBNA 2B protein. In the present work 12 type A isolates and 8 type B isolates have been compared for their ability to transform resting human B cells in vitro into permanent lymphoblastoid cell lines. Although the kinetics of initial focus formation was not markedly dependent upon the EBNA 2 type of the transforming virus, on subsequent passage type A virus-transformed cells (type A transformants) yielded cell lines much more readily than did type B transformants. Direct comparison between the two types of transformant revealed clear differences in several aspects of growth phenotype. Compared with type A transformants, cell lines established with type B virus isolates consistently displayed an unusual growth pattern with poor survival of individual cells shed from lymphoblastoid clumps, a lower growth rate and a greater sensitivity to seeding at limiting dilutions, and a significantly lower saturation density that could not be corrected by supplementation of the medium with culture supernatant containing B-cell growth factors. This is the first direct evidence that, in EBV transformed B-cell lines, the EBNA 2 protein plays a continuing role in determining the cellular growth phenotype. PMID- 3033262 TI - Visna virus exhibits a complex transcriptional pattern: one aspect of gene expression shared with the acquired immunodeficiency syndrome retrovirus. AB - A complex pattern of gene expression was found for visna virus in a highly permissive cell culture system in vitro. In addition to the genomic RNA (9.4 kilobases [kb]), five other mRNAs were detected. The three large RNA transcripts (5.0 kb and a doublet at 4.3 kb) arise by a single splicing event joining 5' sequences to sequences located at positions 3' to the pol gene. The two smallest transcripts (1.8 and 1.5 kb) are at least doubly spliced mRNAs which contain sequences derived from the 5' end of the genome, the region between the pol and env genes, and 3' terminal sequences. In addition to this complex pattern of transcription, the mRNAs appear to be regulated temporally. The 1.5-kb mRNA appears 6 h later than the other transcripts. The significance of this complex pattern of gene expression in the unique aspects of the lentivirus life cycle and pathogenesis is considered. PMID- 3033263 TI - A mutated membrane protein of vesicular stomatitis virus has an abnormal distribution within the infected cell and causes defective budding. AB - Two temperature-sensitive (ts) mutants of the M protein of vesicular stomatitis virus (tsG31 and tsG33) are defective in viral assembly, but the exact nature of this defect is not known. When infected cells are switched from nonpermissive (40 degrees C) to permissive (32 degrees C) temperatures in the presence of cycloheximide, tsG33 virus release increased by 100-fold, whereas tsG31 release increased only by 10-fold. Thus, the tsG33 defect is more reversible than that of tsG31. Therefore, we investigated how the altered synthesis and cellular distribution of tsG33 M protein correlates with the viral assembly defect. At 32 degrees C tsG33 M protein is stained diffusely in the cell cytoplasm and later at the budding sites. In contrast, at 40 degrees C the mutant M protein formed unusual aggregates mostly located in the perinuclear regions of virus-infected cells and partially colocalized with G protein in this region. In temperature shift-down experiments, M can be disaggregated and used to some extent for nucleocapsid coiling and budding, which correlates with the virus titer increase. M aggregates also formed after shift-up from 32 to 40 degrees C, indicating a complete dependence of M aggregation on the temperature. Biochemical analysis with sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting revealed that at 40 degrees C M protein is detected exclusively in pellet fractions (nuclear and cytoskeleton components), whereas at 32 degrees C M protein is mainly in the cytoplasmic soluble fractions. Furthermore, when the temperature is raised from 32 to 40 degrees C, the distribution of M protein tends to shift from the soluble to the pellet and cytoskeletal fractions. Electron micrographs of immunoperoxidase-labeled M protein showed that at 40 degrees C M aggregates are often associated with the outer nuclear membranes as well as with vesicular structures. No nucleocapsid coiling was observed in these cells, whereas coiling and budding were seen at 32 degrees C in cells where M protein was partly associated with the plasma membrane. We suggest that the tsG33 M protein mutation may produce a reversible conformational alteration which causes M protein to aggregate at 40 degrees C, therefore inhibiting the proper association of M protein with nucleocapsids and budding membranes. PMID- 3033264 TI - Nucleotide sequence of a cDNA clone carrying the glycoprotein gene of infectious hematopoietic necrosis virus, a fish rhabdovirus. AB - The nucleotide sequence of the mRNA encoding the glycoprotein of infectious hematopoietic necrosis virus was determined from a cDNA clone containing the entire coding region. The G-protein cDNA is 1,609 nucleotides long (excluding the polyadenylic acid) and encodes a protein of 508 amino acids. The predicted amino acid sequence was compared with that of the glycoprotein of the Indiana and New Jersey serotypes of vesicular stomatitis virus and with the glycoprotein of rabies virus, using a computer program which determined optimal alignment. An amino acid identity of approximately 20% was found between infectious hematopoietic necrosis virus and the two vesicular stomatitis virus serotypes and between infectious hematopoietic necrosis virus and rabies virus. The positions and sizes of the signal sequence and transmembrane domain and the possible glycosylation sites were determined. PMID- 3033265 TI - Tissue-specific replication of Friend and Moloney murine leukemia viruses in infected mice. AB - We have studied the replication of ecotropic murine leukemia viruses (MuLV) in the spleens and thymuses of mice infected with the lymphocytic leukemia-inducing virus Moloney MuLV (M-MuLV), with the erythroleukemia-inducing virus Friend MuLV (F-MuLV), or with in vitro-constructed recombinants between these viruses in which the long terminal repeat (LTR) sequences have been exchanged. At 1 week after infection both the parents and the LTR recombinants replicated predominantly in the spleens with only low levels of replication in the thymus. At 2 weeks after infection, the patterns of replication in the spleens and thymuses were strongly influenced by the type of LTR. Viruses containing the M MuLV LTR exhibited a remarkable elevation in thymus titers which frequently exceeded the spleen titers, whereas viruses containing the F-MuLV LTR replicated predominantly in the spleen. In older preleukemic mice (5 to 8 weeks of age) the structural genes of M-MuLV or F-MuLV predominantly influenced the patterns of replication. Viruses containing the structural genes of M-MuLV replicated efficiently in both the spleen and thymus, whereas viruses containing the structural genes of F-MuLV replicated predominantly in the spleen. In leukemic mice infected with the recombinant containing F-MuLV structural genes and the M MuLV LTR, high levels of virus replication were observed in splenic tumors but not in thymic tumors. This phenotypic difference suggested that tumors of the spleen and thymus may have originated by the independent transformation of different cell types. Quantification of polytropic MulVs in late-preleukemic mice infected with each of the ecotropic MuLVs indicated that the level of polytropic MuLV replication closely paralleled the level of replication of the ecotropic MuLVs in all instances. These studies indicated that determinants of tissue tropism are contained in both the LTR and structural gene sequences of F-MuLV and M-MuLV and that high levels of ecotropic or polytropic MuLV replication, per se, are not sufficient for leukemia induction. Our results further suggested that leukemia induction requires a high level of virus replication in the target organ only transiently during an early preleukemic stage of disease. PMID- 3033266 TI - Map position and nucleotide sequence of the gene for the large structural phosphoprotein of human cytomegalovirus. AB - Human cytomegalovirus particles contain a phosphoprotein of 150,000 (pp150) apparent molecular weight in their matrix; the protein appears particularly reactive in Western blot analyses with human antisera. The gene for pp150 was mapped by screening a bacteriophage lambda gt11 cDNA expression library with monospecific rabbit antisera. Subsequent hybridization of cDNA with cosmid and plasmid clones containing the human cytomegalovirus strain AD169 genome mapped the gene to HindIII fragments J and N. The gene is transcribed into a late 6.2 kilobase RNA. The nucleotide sequence of this region was determined, and a transcription initiation site and two polyadenylation sites of an abundant transcript were located by primer extension and nuclease protection experiments. The reading frame for pp150, deduced from computer analyses, gives rise to a polypeptide of 1,048 amino acids in length; protein secondary structure analysis revealed multiple beta-pleated sheets in hydrophilic clusters, providing a possible explanation for the immunogenic properties of the polypeptide. PMID- 3033267 TI - Vesicular stomatitis virus in Drosophila melanogaster cells: lack of leader RNA transport into the nuclei and frequent abortion of the replication step. AB - In cultured Drosophila melanogaster cells, vesicular stomatitis virus (VSV) establishes a persistent, noncytopathic infection. No inhibition of host macromolecular synthesis occurs. We studied the synthesis of VSV plus-strand leader RNA, which may be directly involved in vertebrate host synthesis shut-off. Leader RNA accumulated in Drosophila cell cytoplasm, but in low amounts, it was either free or associated to structures larger than the leader RNA-N protein complexes found in vertebrate cells. Only a few leader RNA copies migrated into the cell nucleus; no increase of this transport was observed at any time during the virus cycle. Viral RNAs complementary to the 3' end of the genome and ranging in size from the leader to several hundred nucleotides were found to accumulate in Drosophila cell cytoplasm. Their synthesis was inhibited in the presence of cycloheximide, which blocks all protein synthesis and VSV replication. Correlation between the absence of VSV cytopathogenicity in Drosophila cells and the lack of leader RNA transport into their nuclei is discussed, as well as the possible relationship between the restriction of viral synthesis and the frequent initiation of an abortive replication step. PMID- 3033268 TI - Nucleotide sequence and transcript organization of a region of the vaccinia virus genome which encodes a constitutively expressed gene required for DNA replication. AB - A vaccinia virus (VV) gene required for DNA replication has been mapped to the left side of the 16-kilobase (kb) VV HindIII D DNA fragment by marker rescue of a DNA- temperature-sensitive mutant, ts17, using cloned fragments of the viral genome. The region of VV DNA containing the ts17 locus (3.6 kb) was sequenced. This nucleotide sequence contains one complete open reading frame (ORF) and two incomplete ORFs reading from left to right. Analysis of this region at early times revealed that transcription from the incomplete upstream ORF terminates coincidentally with the complete ORF encoding the ts17 gene product, which is directly downstream. The predicted proteins encoded by this region correlate well with polypeptides mapped by in vitro translation of hybrid-selected early mRNA. The nucleotide sequences of a 1.3-kb BglII fragment derived from ts17 and from two ts17 revertants were also determined, and the nature of the ts17 mutation was identified. S1 nuclease protection studies were carried out to determine the 5' and 3' ends of the transcripts and to examine the kinetics of expression of the ts17 gene during viral infection. The ts17 transcript is present at both early and late times postinfection, indicating that this gene is constitutively expressed. Surprisingly, the transcriptional start throughout infection occurs at the proposed late regulatory element TAA, which immediately precedes the putative initiation codon ATG. Although the biological activity of the ts17-encoded polypeptide was not identified, it was noted that in ts17-infected cells, expression of a nonlinked VV immediate-early gene (thymidine kinase) was deregulated at the nonpermissive temperature. This result may indicate that the ts17 gene product is functionally required at an early step of the VV replicative cycle. PMID- 3033269 TI - Identification of gp350 as the viral glycoprotein mediating attachment of Epstein Barr virus (EBV) to the EBV/C3d receptor of B cells: sequence homology of gp350 and C3 complement fragment C3d. AB - The major Epstein-Barr virus (EBV) envelope glycoprotein, gp350, was purified from the B95-8 cell line and analyzed for its ability to mediate virus attachment to the isolated EBV/C3d receptor (CR2) of human B lymphocytes. Purified gp350 and EBV, but not cytomegalovirus, exhibited dose-dependent binding to purified CR2 in dot blot immunoassays. Binding was inhibited by certain monoclonal antibodies to CR2 and to gp350. Liposomes bearing incorporated gp350 bound to CR2-positive B cell lines but not to CR2-negative lines. Liposome binding was also inhibited by the OKB7 anti-CR2 monoclonal antibody. A computer-generated comparison of the deduced gp350 amino acid sequence with that of the human C3d complement fragment revealed two regions of significant primary sequence homology, a finding which suggests that a common region on these two unrelated proteins may be involved in CR2 binding. PMID- 3033270 TI - Simian immunodeficiency virus induces expression of class II major histocompatibility complex structures on infected target cells in vitro. AB - The human immunodeficiency virus (HIV) and the closely related simian immunodeficiency virus (SIV) induce profound immune dysfunction in primate species. The present studies show that cell populations infected in vitro with SIV exhibit increases in major histocompatibility complex (MHC) class II antigen expression. Cell lines chronically infected with both the monkey and human viruses express substantially more MHC class II but not more lineage-restricted or activation antigens on their membranes than do uninfected cell lines. Furthermore, 2'-deoxy-5-iodouridine increased MHC class II antigen expression on SIV-infected cell lines in parallel with increased expression of viral antigens. MHC class II induction does not appear to be mediated through the production of a soluble factor, such as gamma interferon, by SIV-infected cells. Interestingly, studies of the kinetics of antigen expression by cell lines after SIV infection indicate that the induction of MHC class II structures is a late event. Immunoelectron microscopy revealed that MHC class II antigen is expressed not only on the surfaces of the SIV-infected cells but also on the envelope of virus particles derived from those cells. MHC antigen expression on virus-infected cells and the expression of those determinants by the virus may play a role in the pathogenesis of acquired immunodeficiency syndrome and the autoimmune abnormalities observed in HIV-infected individuals. PMID- 3033272 TI - Owl monkey astrocytoma cells in culture spontaneously produce infectious JC virus which demonstrates altered biological properties. AB - A tumor cell suspension of an explanted JC virus (JCV)-induced owl monkey glioblastoma was inoculated intracranially into four recipient juvenile owl monkeys. Twenty-eight months following inoculation one owl monkey developed a glioblastoma, which was explanted into tissue culture. DNA from both the tumor tissue and tumor cells in culture hybridized to a JCV DNA probe by Southern analysis, indicating that free, as well as integrated, viral DNA may be present. At the time of the second culture passage, viral JCV DNA was extracted from these cells and cloned into a plasmid vector. Nucleotide sequencing of the regulatory region of the cloned DNA demonstrated homology with the prototype Mad-1 strain of JCV and revealed a 19-base-pair deletion in the second 98-base-pair tandem repeat that eliminated a second TATA box. This deletion is characteristic of the Mad-4 strain of JCV, which is highly neurooncogenic. By the third culture passage, 100% of the cells were T-antigen positive. Approximately one-third of the cells in culture hybridized to a biotinylated JCV DNA probe when in situ hybridization was used, a technique that only detects high-copy-number of replicating viral sequences. By the culture passage 5 and continuing through culture passage 14, viable JC virions could be recovered. The T protein synthesized by this virus, now termed JCV-586, differed from both the Mad-1 and Mad-4 strains in that it formed a stable complex with the cellular p53 protein in the tumor cells. Also, the JCV-586 T protein reacted to several monoclonal antibodies made to the simian virus 40 T protein that were not recognized by either the Mad-1 or Mad-4 strains. PMID- 3033271 TI - Herpes simplex virus infection generates large tandemly reiterated simian virus 40 DNA molecules in a transformed hamster cell line. AB - When the simian virus 40 (SV40)-transformed Syrian hamster cell line Elona is infected with herpes simplex virus type 1, an excessive amplification of SV40 specific DNA sequences occurs. Analysis of total DNA from herpes simplex virus infected cells revealed that amplified DNA sequences were present predominantly in a high-molecular-weight form, consisting of a tandem array of many unit-length SV40 DNA molecules. Repeat units of amplified DNA were found to be very similar to standard SV40 DNA as was shown by restriction analyses, except for a small deletion close to the origin of replication, which could also be detected in the chromosomal DNA of uninfected cells. A procedure, devised for selective enrichment of amplified SV40 DNA molecules from the bulk of cellular and herpesviral DNA, allowed molecular cloning of single repeat units and nucleotide sequence analysis of the relative genomic region. PMID- 3033273 TI - Regions of poliovirus protein VP1 produced in Escherichia coli induce neutralizing antibodies. AB - Poliovirus type 1 cDNA was prepared from viral RNA encoding the VP1 capsid region of the virus by using a specific DNA primer and was cloned in Escherichia coli. DNA fragments corresponding to VP1 amino acid positions 129 to 302 (pPM5k3), 52 to 302 (pPMhae3), and 24 to 129 (pPMDxba) were incorporated into plasmid vectors designed to express Trp LE-poliovirus VP1 fusion proteins under the control of the inducible tryptophan promoter-operator system. Induction of bacterial cultures containing the plasmids resulted in the production of fusion proteins which accounted for 21% (pPMhae3), 68% (pPM5k3), and 27% (pPMDxba) of the total cell protein. The proteins were purified, and each reacted with polyclonal antibodies raised against intact virions as measured by an enzyme-linked immunosorbent assay. The sera from rabbits immunized with the bacterially produced fusion proteins pPMDxba and pPMhae3 contained poliovirus-neutralizing antibodies. PMID- 3033274 TI - Construction of a genetic switch for inducible trans-activation of gene expression in eucaryotic cells. AB - The cotransfection of selectable marker genes and the gene for the nonstructural proteins NS1 and NS2 of the autonomous parvovirus H-1 failed to produce cell lines that constitutively expressed NS1. A plasmid, pP38NS1cat, was constructed that expressed the NS1-NS2 gene from the H-1 P38 coat protein promoter in place of the natural P4 promoter. The P38 promoter is constitutively weak and is trans activated by NS1. Stable cell lines were isolated that contained pP38NS1cat that was constitutively silent, but inducible with exogenous NS1 by superinfection or by treatment with sodium butyrate. The cells that were induced for this self stimulatory genetic circuit did not remain in the culture, suggesting that expression of NS1-NS2 is cytotoxic or that the expression is not sustained. The properties of these cell lines and an example of the construction of a cell line inducible for expression of the viral coat protein gene and the bacterial gene for chloramphenicol acetyltransferase (cat) are described. PMID- 3033275 TI - Construction of viable deletion and insertion mutants of the Sabin strain of type 1 poliovirus: function of the 5' noncoding sequence in viral replication. AB - A number of deletion and insertion sequences were introduced into the 5' noncoding sequence (742 nucleotides long) of the genome of the Sabin strain of type 1 poliovirus by using an infectious cDNA clone of the virus strain. The genomes of all three poliovirus serotypes contained highly homologous sequences (nucleotide positions 509 to 639) as well as highly variable sequences (positions 640 to 742) in the 5' noncoding region. The viability of mutant viruses was tested by transfecting mutant cDNA clones into African green monkey kidney cells and then estimating the plaque sizes displayed on the cells. The results suggested that the highly variable sequence next to the VP4 coding region did not play an important role, at least in the in vitro culture system used, that the loci of highly conserved nucleotide sequences were not always expected to be the genome regions essential for viral replication, that the sequence between positions 564 and 599 carried genetic information to maintain the efficiency of certain steps in viral replication, and that the sequence between positions 551 to 563 might play an essential role in viral replication. Four-base deletion or insertion mutations were introduced into relatively variable sequences in the genome region upstream of position 509. The results suggest that variable sequences do not always indicate that the corresponding genome regions are less important. Apparent revertants (large-plaque variants) were easily generated from one of the viable mutants with the small-plaque phenotype. The determination of nucleotide sequences of the revertant genomes revealed the second mutation site. The results suggested that the different loci at around positions 200 and 500 might specifically interact with each other. This interaction may result in the formation of a functional structure that influences the efficiency of certain steps in the viral replication. PMID- 3033276 TI - Synthesis and immunogenicity of the rotavirus major capsid antigen using a baculovirus expression system. AB - Rotaviruses are the major pathogens that cause life-threatening diarrhea in young children and animals. We inserted a simian rotavirus SA11 gene 6 cDNA into the genome of the baculovirus Autographa californica nuclear polyhedrosis virus adjacent to the strong polyhedrin promoter. The major capsid antigen (VP6) was expressed in high yields (20 to 150 micrograms/10(6) cells) when Spodoptera frugiperda cells were infected with baculovirus recombinants containing SA11 gene 6 inserts. Reactivity with monospecific polyclonal and monoclonal antibodies suggested that VP6, expressed intracellularly or found in the media, maintained native antigenic determinants. VP6 purified from the media from infected cells also possessed a native oligomeric structure, was immunogenic in guinea pigs, and was able to spontaneously assemble into morphologic subunits. Antisera from immunized guinea pigs failed to neutralize virus in plaque reduction assays, but detected homologous and heterologous rotavirus strains when tested by immunofluorescence, immunoprecipitation, and enzyme-linked immunosorbent assays. PMID- 3033277 TI - Sequences of the Epstein-Barr Virus (EBV) large internal repeat form the center of a 16-kilobase-pair palindrome of EBV (P3HR-1) heterogeneous DNA. AB - We have previously characterized several genomic rearrangements of Epstein-Barr virus (EBV) DNA contained in one of the defective EBV genomes harbored by the P3HR-1 (HR-1) line (H. B. Jenson, M. S. Rabson, and G. Miller, J. Virol. 58:475 486, 1986). One recombinant clone of heterogeneous DNA (het DNA) from this defective genome is an EcoRI fragment of 16 kilobase pairs (kbp) which is a palindrome. DNA digestion fragments specific for the center of this palindrome were present in cells which contained het DNA but not in cells which lacked het DNA. Thus, the palindrome was not an artifact of DNA cloning. The organization of the center of this palindrome was studied by DNA sequencing. The comparable region of the parental HR-1 genome was also studied by DNA sequencing. The central 3,495 base pairs (bp) of the palindrome were composed of sequences derived exclusively from internal repeat 1 of EBV, represented by BamHI W fragment. At each end of the central 3,495 hp was a symmetrical recombination with sequences of BamHI-Z, located more than 50 kbp away on the standard EBV genome. The central 3,495 bp were composed of an unduplicated 341 bp flanked by two perfect palindromic repeats of 1,577 bp. The 341-bp unique region was a portion of a 387-bp region of standard HR-1 BamHI-W which was identical to the central 387 bp of the palindrome. This central 387-bp region contained numerous stretches of dyad symmetry capable of forming a large stem-and-loop structure. The palindromic rearrangement had created two novel open reading frames in het DNA derived from standard HR-1 BamHI-W sequences. These two het DNA open reading frames had different amino termini but identical carboxy termini derived from the large open reading frame in standard HR-1 BamHI-W (HR-1 BWRF1). The BamHI-W sequences found in het DNA did not include either the TATA box of standard HR-1 BamHI-W or the exons which are present in the potentially polycistronic latent mRNAs encoding EBV nuclear antigens. These marked alterations in genomic structure may relate to the unique biologic properties of virus stocks containing het DNA by creation of new polypeptides or by formation or deletion of regulatory or functional signals. PMID- 3033278 TI - Analysis of the complete nucleotide sequence of the picornavirus Theiler's murine encephalomyelitis virus indicates that it is closely related to cardioviruses. AB - Theiler's murine encephalomyelitis viruses (TMEV) are naturally occurring enteric pathogens of mice which constitute a separate serological group within the picornavirus family. Persistent TMEV infection in mice provides a relevant experimental animal model for the human demyelinating disease multiple sclerosis. To provide information about the TMEV classification, genome organization, and protein processing map, we determined the complete nucleotide sequence of the TMEV genome and deduced the amino acid sequence of the polyprotein coding region. The RNA genome, which is typical of the picornavirus family, is 8,098 nucleotides long. The 5' untranslated region is 1,064 nucleotides long (making it the longest in the picornavirus family after the aphthoviruses) and lacks a poly(C) tract. Computer-generated comparison of the 5' and 3' noncoding regions and polyprotein revealed the highest level of nucleotide and predicted amino acid identity between the TMEV and the cardioviruses encephalomyocarditis virus (EMCV) and Mengo virus. The TMEV polyprotein, which appears to be processed like EMCV since the amino acids flanking the putative proteolytic cleavage sites have been conserved, begins with a short leader peptide followed by 11 other gene products in the standard L-4-3-4 picornavirus arrangement. Because of these similarities, we propose that the TMEV be grouped with the cardioviruses. However, since TMEV and EMCV have different biophysical properties and show no cross-neutralization, they most likely belong in a separate cardiovirus subgroup. PMID- 3033280 TI - Structural characteristics of the Corynebacterium lilium bacteriophage CL31. AB - Bacteriophage CL31 was isolated on a Corynebacterium lilium strain. Out of 30 strains tested, only CL31 was able to form plaques on Corynebacterium glutamicum ATCC 13287, Brevibacterium lactofermentum ATCC 21086, and Arthrobacter sp. strain SI55, but at a very low frequency. This phage belongs to group B of Bradley's classification (D. E. Bradley, Bacteriol. Rev. 31:230-314; 1967). Its head is 53 nm in diameter, and its tail is 396 nm in length. The phage capsid contains three major proteins, of 12.5, 29.0, and 37.0 kilodaltons, and five minor ones (23.9, 26.0, 27.0, 40.0, and 55.4 kilodaltons). CL31 DNA is a linear molecule of 48 kilobases with cohesive ends. Restriction mapping was performed for endonucleases BglII, EcoRI, SalI, and KpnI. The expression of CL31 genes in Escherichia coli was studied by the maxicell technique; 12 different proteins were detected. PMID- 3033279 TI - Native and recombinant herpes simplex virus type 1 envelope proteins induce human immune T-lymphocyte responses. AB - The abilities of whole herpes simplex virus type 1 (HSV-1) antigen (HSV-ag) and purified HSV-1 native and recombinant envelope proteins to stimulate in vitro T lymphocyte responses were compared in patients with recurrent herpes labialis. Immunochemically purified preparations of native glycoproteins B, C, and D (ngB, ngC, ngD) from cultured HSV-1 as well as expressed recombinant plasmid preparations of gD (rgD-1t, rgD-45K) elicited lymphocyte proliferation (LT) and production of gamma interferon (IFN-gamma) and interleukin-2 (IL-2) only in seropositive individuals. The IFN-gamma induced by rgD-1t correlated with the time to the next herpetic lesion in 19 volunteers followed to recurrence (r = 0.69, P less than 0.008), although the magnitude and frequency of LT and IFN gamma responses were lower with either recombinant or native purified antigens than with the whole-virus antigen. Combinations of ngB plus ngD or ngB plus ngC plus ngD stimulated more IFN-gamma, equivalent to whole-virus-antigen responses. Recombinant-derived human IL-2 also specifically increased LT and IFN-gamma responses in antigen-driven cultures. ngD stimulated IL-2 and LT responses similar to those of whole-virus antigen and higher than those of ngC. HSV-ag and ngB induced significantly higher titers of total IFN than could be accounted for by IFN-gamma; this was not seen for the other antigens, which induced only IFN gamma. HSV-ag-driven Leu 2a-, plastic-nonadherent blood cells, unlike whole peripheral blood mononuclear cells, showed evidence of an increase and then a decline in the frequency of HSV-responsive cells after a lesion recurrence. These studies suggest that HSV-1 envelope proteins are capable of stimulating an immune T-helper-cell response which is associated with the prevention of human herpes simplex lesion recurrence. Although the whole virus probably contains additional important antigens, increasing concentrations or combinations of certain purified glycoproteins or the addition of nonspecific enhancers of T-lymphocyte function can drive in vitro immune responses to the same level as the complete set of viral antigens. PMID- 3033281 TI - Cyclic AMP specifically blocks proliferation of rat 3T3 cells transformed by polyomavirus. AB - Elevated exogenous and intracellular levels of cyclic AMP could totally block proliferation of polyomavirus (PyV) transformants derived from rat 3T3 cells without affecting proliferation of normal cells or simian virus 40 (SV40)-induced transformants. Concanavalin A (ConA) had the opposite effect; it could totally block proliferation of both normal cells and SV40 transformants but reduced proliferation of PyV transformants only twofold. Adenylate cyclase was threefold less active in membranes of PyV transformants, and the number of ConA receptors was similar to that of normal cells. Proliferating PyV transformants contained threefold less cyclic AMP than did proliferating SV40 transformants. The sensitivity to cyclic AMP did not correlate with the degree of transformation: cells transformed by Rous sarcoma virus and tumor cells derived from SV40 transformants were not sensitive to cyclic AMP. The differential effect of cyclic AMP and ConA on proliferation was probably due to the activity of an intact middle t protein. The presence of both large T and small t together with middle t was also required for cyclic AMP sensitivity. PMID- 3033282 TI - Human neonatal lymphocytes immortalized after microinjection of Epstein-Barr virus DNA. AB - Epstein-Barr virus (EBV) is a highly efficient acute transforming agent in human cells, provided that the intact virus is used. To investigate the ability of viral DNA alone to transform cells, we introduced the EBV genome into human lymphocytes. After microinjection of EBV DNA into neonatal B lymphocytes, we established a cell line that in early passages contained multiple viral fragments. This cell line retained sequences from the short, unique (Us) region of the EBV genome and sequences from EcoRI-E. The viral sequences were not expressed; however, the cells expressed a 2.3-kilobase polyadenylated message homologous to the c-fgr oncogene, a cellular locus believed to be activated by EBV infection [M. S. C. Cheah, T. J. Ley, S. R. Tronick, and K. C. Robbins, Nature (London) 319:238-240.]. The cell line was monoclonal with rearrangement at the immunoglobulin locus and had a reciprocal translocation t(1;7)(p34;q34) and a deletion of sequences within the locus for the beta chain of the T-cell receptor. The close proximity of the translocation to the chromosomal loci for c-fgr on chromosome 1 and the T-cell receptor beta chain on chromosome 7 suggests that structural alteration of these genes was critical to this transformation event. PMID- 3033283 TI - Multiple tandemly repeated binding sites for cellular nuclear factor 1 that surround the major immediate-early promoters of simian and human cytomegalovirus. AB - We show that the large DNA genomes of human and simian cytomegaloviruses (HCMV and SCMV, respectively) each contain multiple binding sites for purified cellular nuclear factor 1 (NF1) protein. Examination of the major immediate-early (IE) gene region in the HindIII H fragment of SCMV (Colburn) by filter binding assays showed that it competed 45-fold better than the single adenovirus type 2 binding site for NF1 protein and that it contained at least two distinct binding loci. Direct DNase I footprinting analyses of the 5' upstream locus detected at least 20 adjacent NF1-binding sites located between positions -600 and -1300 relative to the IE94 mRNA start site. DNA sequence analysis of the region revealed a conserved consensus NF1 recognition element (T)TGG(C/A)N5GCCAA embedded within each of 23 highly diverged 30-base-pair tandem repeats, together with a second downstream cluster of five consensus NF1-binding sites between positions +470 and +570 in the large first intron. Two separate NF1-binding loci were also found in the equivalent IE68 gene of HCMV(Towne) DNA, but in this case the DNA sequence and competition filter binding experiments indicated a maximum of only four to five consensus binding sites encompassing the promoter-enhancer region. In transient expression assays, neither the isolated upstream IE94 tandem repeats nor a synthetic single-copy consensus NF1-binding site acted as transcriptional cis activators or enhancers when placed adjacent to the simian virus 40 minimal early region promoter. We conclude that the large and complex 5' upstream promoter-regulatory region for the SCMV IE94 gene comprises two distinct domains. The previously described four sets of 13- to 18-base-pair interspersed repeat elements between -55 and -580 provide most of the high basal transcriptional strength, whereas the arrangement of further upstream tandemly repeated NF1 binding sites may contribute significantly to the expanded biological host range for expression of SCMV IE94 compared with HCMV IE68. PMID- 3033284 TI - Expression of the bovine leukemia virus X region in virus-infected cells. AB - Bovine leukemia virus, like its closest relatives the human T-cell leukemia virus types I and II, contains a 1.8-kilobase X region between the env gene and the 3' long terminal repeat. In this communication, we report the detection and characterization of a subgenomic mRNA from which this X region is presumably translated. This mRNA was produced by a complex splicing mechanism which resulted in juxtaposition of the 5' end of the env gene and the two overlapping X-region open reading frames. Translation of this mRNA could yield at least two distinct proteins depending on which initiation codon is used. Detection of the protein encoded by the BLV X-region long open reading frame has been reported (N. Sagata, J. Tsuzuku-Kawamura, M. Nagayoshi-Aida, F. Shimizu, K.-I. Imagawa, and Y. Ikawa, Proc. Natl. Acad. Sci. USA 82:7879-7883, 1985). Using synthetic peptide antisera, we detected a protein encoded by the short open reading frame in virus-infected cells. The protein migrated in sodium dodecyl sulfate-polyacrylamide gels with an apparent molecular weight of 19,000. It is a nuclear phosphoprotein. PMID- 3033285 TI - Differences among human immunodeficiency virus strains in their capacities to induce cytolysis or persistent infection of a lymphoblastoid cell line immortalized by Epstein-Barr virus. AB - Four strains of human immunodeficiency virus (HIV) manifest consistent differences in biologic behavior after infection of the X50-7 line of human umbilical cord lymphocytes immortalized by Epstein-Barr virus (EBV). Some dilutions of the first strain examined, human T-cell lymphotropic virus type III B, which is derived from a pool of patient isolates propagated in H9 cells, caused transient cytopathic effects (CPE) followed by recovery of a subpopulation of X50-7 cells which became virus carrier cultures. Other dilutions of the same virus stock completely lysed X50-7 cells. Two other strains, RF2 and YW, both from individual patients with acquired immune deficiency syndrome, always induced complete cytolysis of X50-7 cells at all dilutions which infected the cells. However, RF2 did establish persistent infection of H9 cells. A fourth strain, PH1 MN, from a child with acquired immune deficiency syndrome-related complex, induced only transient CPE in X50-7 and H9 cells, which thereafter always recovered to form carrier cultures. For all four strains, the dilutions of HIV stocks which caused CPE corresponded to dilutions which resulted in the detection of HIV polypeptides by immunoblot. Cytolysis in HIV-infected X50-7 cells was accompanied by a decrease in the amount of EBV nuclear antigen; however, HIV infection did not induce EBV replication. Thus CPE in X50-7 cells is due to replication of HIV per se and not to activation of EBV. The observations indicate that there are differences in the cytolytic properties of HIVs and that these differences are influenced by the target cell. PMID- 3033286 TI - Mutations within the proteolytic cleavage site of the Rous sarcoma virus glycoprotein that block processing to gp85 and gp37. AB - We have investigated the specificity of the proteolytic cleavage of the Rous sarcoma virus glycoprotein precursor by introducing two mutations into the putative cleavage region (Arg-Arg-Lys-Arg). We show that neither a deletion of the cleavage sequence nor a glutamic acid for lysine substitution altered intracellular transport or surface expression of the env gene products. However, both the four-amino-acid deletion and the glutamic acid substitution block processing of the env precursor. Susceptibility of the glutamic acid-substituted env precursor to proteases indicated that tertiary protein structure was unaffected. While inhibitor experiments suggested that more than one endopeptidase might be capable of mediating the proteolytic cleavage, the results presented here point to the presence in the Golgi apparatus of a novel endopeptidase, required for retroviral glycoprotein cleavage, that has a high specificity for lysine-containing peptides. PMID- 3033287 TI - Replicative cis-advantage of polyomavirus regulatory region mutants in different murine cell lines. AB - To determine the relative growth advantages of polyomavirus regulatory region mutants selected from Friend leukemic and neuroblastoma cells persistently infected with polyomavirus A2 wild type, different murine cell lines, some of which are capable of further differentiation in vitro, were used as hosts in mixed infection and transfection. The tests allowed the determination, through the measurement of cis-advantage in replication, of the most effective polyomavirus regulatory region constitutions for a given cell line and, in some cases, for specific stages of cell differentiation. Furthermore, different domains of the viral regulatory region were shown to have different effects- positive, neutral, or negative--depending on the host analyzed. PMID- 3033288 TI - Subtyping of European foot-and-mouth disease virus strains by nucleotide sequence determination. AB - The VP1-coding regions of foot-and-mouth disease virus strains from 18 recent European outbreaks and of 9 strains isolated more than 20 years ago and used in part as vaccines were determined by direct cDNA sequencing. Comparison of the sequences revealed that most of the isolated outbreak viruses are closely related to the vaccine strains used. Isolates from the Italian epizootic of 1984 to 1985 correspond, for example, to the vaccine strain A5 Parma 62; the outbreak in 1984 in Bernbeuren, Federal Republic of Germany, was induced by A5 Allier 60; outbreaks in 1982 in Funen, Denmark, and in Murchin, German Democratic Republic, were caused by O1 Lausanne 65. Viruses isolated during the 1983 Iberian epizootic show a close relationship to the vaccine strain A5 Allier 60 but were probably derived from another not yet identified vaccine strain from Spain. Only two minor outbreaks in the Federal Republic of Germany, A Aachen in 1976 and O Wuppertal in 1982, did not correspond to the classical European strains but were obviously introduced from outside. We suggest that nucleotide sequence analysis should be used as a standard method of diagnosis, because when compared with other techniques it more clearly reveals the origin and course of epizootics and offers the possibility of preventing further outbreaks. PMID- 3033289 TI - Transcriptional trans-activation by the human papillomavirus type 16 E2 gene product. AB - We identified a conditional transcriptional enhancer in the long control region (LCR) of human papillomavirus type 16 (HPV-16). This conditional enhancer requires activation in trans by a product of the viral early-region open reading frames (ORFs). Primer extension analysis of chloramphenicol acetyltransferase RNA isolated from transiently transfected CV-1 cells demonstrated that trans activation of the HPV-16 LCR enhancer operated at the transcriptional level. Mutational analysis of the early ORFs demonstrated that the conditional enhancer of the LCR was trans-activated by the product of the E2 ORF. The E2 gene product of bovine papillomavirus type 1, which can trans-activate the conditional enhancer in the bovine papillomavirus type 1 LCR, was also capable of trans activating the E2-responsive enhancer of HPV-16. The activity of the HPV-16 LCR enhancer was also assayed in two human cervical carcinoma cell lines, HeLa and SiHa, which harbor transcriptionally active, integrated HPV-18 and HPV-16 DNA sequences, respectively. No endogenous E2 or E2-like activity was detected in either cell line. PMID- 3033290 TI - Effect of internal viral sequences on the utility of retroviral vectors. AB - Expression of the human ADA cDNA encoded by the Moloney murine leukemia virus spliced RNA form is enhanced by intron-contained sequences. The presence of sequences corresponding to the viral gag gene in a Moloney murine leukemia virus based vector results in the generation of 10- to 40-fold higher titers of virus. PMID- 3033291 TI - A standardized nomenclature for endogenous mouse mammary tumor viruses. AB - We propose a revised standardized nomenclature for endogenous mouse mammary tumor viruses based on characterization by molecular cloning techniques and genetic segregation data. PMID- 3033292 TI - DNA-binding activity of papillomavirus proteins. AB - We demonstrate DNA binding by papillomavirus (PV) open reading frame (ORF) proteins that correspond to the early transforming and trans-activating (E6 and E2) and late structural regions (L2 and L1) from bovine PV type 1 and human PV types 6b and 16. All PV proteins were synthesized in Escherichia coli and had a common 13-amino-acid leader sequence from the expression vector pRA10. Antibodies have been generated in rabbits against these PV proteins. The PV ORF proteins bind double-stranded DNA, and this activity is demonstrated to be inherent to the PV proteins. DNA-binding activity by PV proteins is optimal at 50 mM NaCl and at pH 7.0. For some PV proteins (e.g., bovine PV type 1 E2), DNA binding is enhanced at a lower pH (pH 6.0) and NaCl concentration (50 to 100 mM). DNA binding is inhibited by the appropriate antibodies. The possible significance of these findings is discussed in relation to the genetic and structural evidence on the function of these ORFs. PMID- 3033294 TI - Resistance/susceptibility to lethal Sendai virus infection genetically linked to a mucociliary transport polymorphism. AB - Linkage was tested between a mucociliary transport polymorphism and resistance/susceptibility to lethal Sendai virus infection in segregant hybrid mice of C57BL/6J and DBA/2J parents. The distribution of paired phenotypes for tracheal mucociliary transport rates and susceptibility to lethal Sendai virus infection in 171 F1 X DBA/2J mice showed strong interaction of the parental phenotypes. PMID- 3033293 TI - Sequences responsible for the altered erythropoietin responsiveness in spleen focus-forming virus strain SFFVP-infected cells are localized to a 678-base-pair region at the 3' end of the envelope gene. AB - Two different strains of Friend spleen focus-forming virus, SFFVP and SFFVA, are known to cause a rapid erythroleukemia. The SFFVP-infected cells can proliferate and differentiate maximally without the addition of the erythroid-specific hormone erythropoietin, whereas the SFFVA-infected cells require erythropoietin for differentiation and for maximum proliferation. We previously reported that a recombinant virus containing sequences from the 3' half of the SFFVP envelope gene and the SFFVP long terminal repeat on an SFFVA background has all of the biological and biochemical characteristics of SFFVP. We are now presenting data on a new recombinant virus to show that only the 3' half of the SFFVP envelope gene is responsible for the differences observed between the two strains. PMID- 3033295 TI - Human papillomavirus type 18 DNA is integrated at a single chromosome site in cervical carcinoma cell line SW756. AB - SW756, a cervical carcinoma cell line, has multiple copies of human papillomavirus type 18 DNA sequences. The integration site of human papillomavirus type 18 DNA was localized by in situ hybridization to chromosome 12 at band q13. This single integration site corresponds to a heritable fragile site, which may have facilitated the integration of the viral DNA. PMID- 3033296 TI - The major human papillomavirus protein in cervical cancers is a cytoplasmic phosphoprotein. AB - In a previous study, the most abundant viral transcript in a human papillomavirus type 16-associated cervical cancer and in a cancer-derived cell line was characterized, and its translation product, the E7 protein, was identified (D. Smotkin and F. O. Wettstein, Proc. Natl. Acad. Sci. USA 68:4680-4684, 1986). Here we show that the E7 protein had a half life of about 1 h and was located in the soluble cytoplasmic fraction. The protein was phosphorylated at serine residues and exhibited a high heterogeneous sedimentation rate in nondenaturing glycerol gradients, suggesting an oligomer formation or association with cellular protein. PMID- 3033297 TI - Continued expression of a poly(A)+ transcript of herpes simplex virus type 1 in trigeminal ganglia of latently infected mice. AB - Radioactively labeled cDNAs were prepared by using as the template poly(A)+ mRNA from trigeminal ganglia of mice latently infected with herpes simplex virus type 1. These cDNAs were used as hybridization probes for Southern blots of cloned herpes simplex virus type 1 DNA fragments. Specific hybridization to fragments from the terminal repetition of the L segment was detected with probes derived from mRNAs obtained as early as 3 weeks and as late as 17 months postinoculation. Fine mapping of the region of hybridization showed that the viral transcripts originated from DNA sequences coding for the immediate-early gene IE-1 (alpha-0). These results indicate that IE-1, or an as yet unidentified gene colinear with it, is continuously expressed during latency. PMID- 3033298 TI - Functional expression of a cloned herpes simplex virus type 1 DNA polymerase gene. AB - A vector which expresses the herpes simplex virus type 1 (HSV-1) (strain 17) DNA polymerase gene was constructed by ligating two separately cloned HSV DNA restriction fragments into an intermediate plasmid and then mobilizing the intact polymerase gene-encoding sequence into a pSV2 derivative. The expression vector (pD7) contains a functional simian virus 40 replication origin and early enhancer promoter upstream from the HSV DNA polymerase-encoding sequence. COS-1 cells transfected with pD7 contained an RNA species, shown by Northern blot analysis to hybridize specifically with an HSV DNA pol probe and to be the same size (4.3 kilobases) as the pol mRNA found in HSV-1-infected COS-1 cells. A genetic complementation test was used to establish that pD7 expresses a functional pol gene product. COS-1 cells transfected with pD7 were able to partially complement the growth defect of an HSV-1 (KOS) temperature-sensitive mutant, tsC7, in the DNA polymerase gene at the nonpermissive temperature. PMID- 3033299 TI - Polyomavirus major capsid protein VP1 is modified by tyrosine sulfuration. AB - Polyomavirus was propagated in primary mouse kidney cell monolayers and 35S sulfate labeled by maintaining the infected cells in serum-free Eagle medium supplemented with 35S-labeled sodium sulfate. Sodium dodecyl sulfate polyacrylamide gel electrophoresis of CsCI gradient-purified 35S-sulfate-labeled virions followed by fluorography indicated that the polyomavirus-coded major capsid protein VP1 incorporated this radiolabel. Two-dimensional gel electrophoresis followed by fluorography revealed 35S-sulfate incorporation into only two of the six VP1 isoelectric species (E and F). Amino acid analysis of 35S sulfate labeled VP1 by enzymatic hydrolysis followed by two-dimensional thin layer electrophoresis revealed the presence of 35S-sulfate-labeled tyrosine-O sulfate. PMID- 3033300 TI - Expression of polyomavirus large T antigen by using a baculovirus vector. AB - A gene encoding the large T antigen of polyomavirus was inserted into the baculovirus Autographa californica nuclear polyhedrosis virus so that gene expression was under the control of the strong, very late polyhedrin gene promoter. Significantly more large T antigen was produced in recombinant virus infected insect cells than was observed in polyomavirus-transformed mouse cells. The insect-derived T antigen exhibited polyomavirus origin-specific DNA binding. The baculovirus expression system provides a convenient source of T antigen for in vitro studies. PMID- 3033301 TI - Intratypic recombination of polioviruses: evidence for multiple crossing-over sites on the viral genome. AB - Intratypic recombinant polioviruses were isolated from cells that were coinfected with two temperature-sensitive (ts) mutants of poliovirus type 1, ts035Gr and ts247. After phenotypic characterization of these recombinants, their proteins were studied by polyacrylamide gel electrophoresis, and their genomes were analyzed by RNase T1 fingerprinting and partial nucleotide sequencing. Segregation of specific phenotypic and biochemical characteristics inherited from the parental viruses demonstrated that crossing-over could occur in at least four distinct regions of the genome. Possible mechanisms for recombination are discussed. PMID- 3033302 TI - Processing of the herpes simplex virus type 2 glycoprotein gG-2 results in secretion of a 34,000-Mr cleavage product. AB - Herpes simplex virus type 2 glycoprotein gG-2 undergoes a cleavage event during its synthesis and processing. The focus of this report is on the detection and fate of the small-molecular-weight component of gG-2, designated the 34K component. In cultures containing the inhibitor monensin, a 31K component accumulated within infected cells. In contrast, the intracellular accumulation of this 31K precursor was not detected in cultures grown in the absence of the inhibitor. However, the 34K component of gG-2 was found in the extracellular culture fluid. The data suggest that the 31K high-mannose cleavage product of gG 2 is further glycosylated and rapidly secreted from herpes simplex virus type 2 infected cells; however, if glycosylation is perturbed, the 31K high-mannose form remains cell associated. PMID- 3033303 TI - Replication of latent Epstein-Barr virus genomes in Raji cells. AB - The replication of the 50 to 60 latent, predominantly extrachromosomal, Epstein Barr virus genomes maintained by the Burkitt-lymphoma-derived Raji cell line was investigated by using a Meselson-Stahl density transfer approach. Samples of DNA isolated from cells cultivated for different periods in bromodeoxyuridine supplemented medium were fractionated according to density, and the distribution of viral and cellular DNAs among the heavy-, hybrid-, and light-density species was quantitated. The results indicate that the majority of latent Epstein-Barr virus DNA plasmids each replicate once during the cell cycle. PMID- 3033304 TI - Structure and function of the transcriptional control region of nonpassaged BK virus. AB - We compared the nucleotide sequence in the transcriptional control region of BK virus isolates cloned directly from human urine (BK-WW) with that of prototype BK virus. BK-WW was found to have a 63-base-pair insertion and only one of the 68 base-pair enhancer repeat elements. In transient expression assays, BK-WW enhancer showed approximately one-half the activity given by the prototype enhancer. PMID- 3033305 TI - In vivo transcription of bacteriophage phi 29 DNA: transcription termination. AB - The main early and late transcription termination sites in vivo in bacteriophage phi 29 DNA were determined by nuclease S1 mapping. Transcription of the phi 29 early genes located at the left end of the viral genome terminated at the very end of the DNA molecule and within the HindIII G fragment of the viral DNA. Transcription termination of the early genes located at the right end of the genome and that of the late viral genes overlapped in a specific region of the phi 29 DNA within the EcoRI D fragment. Stem-loop structures followed by uridine rich tails could be derived close to the 3' ends of early and late mRNAs, suggesting Rho-independent transcription termination in phi 29 DNA. PMID- 3033307 TI - A baculovirus vector can express intron-containing genes. AB - A simjan virus 40 genomic fragment containing the genes coding for the large T and small t antigens was inserted into the genome of the baculovirus Autographa californica nuclear polyhedrosis virus downstream of the strong polyhedrin promoter. Infection of eucaryotic Spodoptera frugiperda (SF9) cells with this recombinant virus produced significant amounts of small t antigen and little or no large T protein. Analysis by Northern blotting and S1 nuclease digestion revealed correct and preferential utilization of the small t splicing signals. PMID- 3033306 TI - Characterization of endonuclease activities in Moloney murine leukemia virus and its replication-defective mutants. AB - To study Moloney murine leukemia virus (M-MulV) proteins associated with the integration of proviral DNA into the host chromosome, we isolated endonuclease activities from purified virion preparations of the wild type and two of its replication mutants. A major endonuclease activity was identified in virions of M MuLV; the enzyme catalyzed nicks in double-stranded DNA in the presence of either Mn2+ or Mg2+ and was stimulated by ATP. The endonuclease nicked DNA adjacent to all four nucleotides with some preference for G and C. The same enzyme, and in comparable amounts, was isolated from two virus replication mutants: dl2905, deficient in the processing of Pr65gag and Pr200gag-pol, and dl50401, deficient for the virus integration function. In the process of these experiments, the residual reverse transcriptase in mutant dl2905 was shown to be the mature size, implying that the uncleaved precursor lacks enzymatic activity. It appears that the major endonuclease activity found in virions of M-MuLV is not encoded by either the gag or pol genes. PMID- 3033309 TI - Evolution of pseudorabies virions containing genomes with an invertible long component after repeated passage in chicken embryo fibroblasts. AB - The genome of pseudorabies virus consists of two components, short (S) and long (L). Only the S component is bracketed by inverted repeats, and only the S component inverts itself relative to the L component, giving rise to two isomeric forms of the genome. An attenuated vaccine strain of pseudorabies virus (Norden), however, has a genome which is found in four isomeric forms (B. Lomniczi, M. L. Blankenship, and T. Ben-Porat, J. Virol. 49:970-979, 1984). To determine the basis for the atypical structure of the genome of the Norden strain, we examined more than 40 field isolates of pseudorabies virus; all contained genomes in which the L component was fixed in only one orientation relative to the S component. Several independently generated vaccine strains which have been passaged extensively in chicken embryos and chicken embryo fibroblast (CEF) cell cultures were also analyzed; they possessed an invertible L component. Furthermore, emergence of pseudorabies virus variants with an invertible L component was observed after passage of the virus in CEF, but not in rabbit kidney or pig kidney, cells. The invertibility of the L component was associated consistently with a translocation of sequences from the left end of the genome to a position next to the inverted repeat sequence of the S component. Three observations indicate that genomes with an invertible L component (and the translocation) have a selective growth advantage over standard pseudorabies virus when grown in CEF. The proportion of virions with such genomes does not increase linearly as would be expected if the translocation events occurred repeatedly, most genomes eventually experiencing the translocation. Instead, after a lag, the proportion of such virions in the population increases relatively rapidly. The genome structures that are generated upon independent passage in CEF of each virion population were relatively homogeneous. Some heterogeneity was observed at relatively early stages of the emergence of the genomes carrying the translocation; at later stages, virions with genomes with a specific size translocation predominated in the virus population. Parallel passages in CEF of the same pseudorabies virus strain resulted in the emergence of populations of virions with genomes with different size translocations. However, in each of the passaged populations of virions the majority of virions had genomes with the same size translocation. The most likely interpretation of these results is that virions with genomes carrying the translocations that emerge upon passage of the virus in CEF have a selective advantage when grown in these cells. PMID- 3033308 TI - DNA-dependent RNA polymerase subunits encoded within the vaccinia virus genome. AB - Antiserum to a multisubunit DNA-dependent RNA polymerase from vaccinia virions was prepared to carry out genetic studies. This antiserum selectively inhibited the activity of the viral polymerase but had no effect on calf thymus RNA polymerase II. The specificity of the antiserum was further demonstrated by immunoprecipitation of RNA polymerase subunits from dissociated virus particles. The presence in vaccinia virus-infected cells of mRNA that encodes the polymerase subunits was determined by in vitro translation. Immunoprecipitable polypeptides with Mrs of about 135,000, 128,000, 36,000, 34,000, 31,000, 23,000, 21,000, 20,000, and 17,000 were made when early mRNA was added to reticulocyte extracts. The subunits were encoded within the vaccinia virus genome, as demonstrated by translation of early mRNA that hybridized to vaccinia virus DNA. The locations of the subunit genes were determined initially by hybridization of RNA to a series of overlapping 40-kilobase-pair DNA fragments that were cloned in a cosmid vector. Further mapping was achieved with cloned HindIII restriction fragments. Results of these studies indicated that RNA polymerase subunit genes are transcribed early in infection and are distributed within the highly conserved central portion of the poxvirus genome in HindIII fragments E, J, H, D, and A. PMID- 3033310 TI - Activation of double-stranded RNA-activated protein kinase in HeLa cells after poliovirus infection does not result in increased phosphorylation of eucaryotic initiation factor-2. AB - Protein kinase activity in general is stimulated at least 5- to 10-fold in ribosomal salt wash preparations from poliovirus-infected HeLa cells compared with those from mock-infected cells. The stimulation of kinase activity is manifested by increased phosphorylation of ribosome-associated polypeptides having approximate molecular weights of 135,000, 120,000, 85,000, 68,000, 65,000, 40,000, 28,000, 25,000, and 21,000. The Mr 68,000 phosphoprotein is structurally identical to the interferon-induced, double-stranded RNA-activated protein kinase (P1) which phosphorylates the alpha subunit of eucaryotic initiation factor-2 (eIF-2). A similar protein of Mr 68,000 is more phosphorylated in poliovirus infected cells than in mock-infected cells. Increased phosphorylation of P1 protein in poliovirus-infected cells, however, does not result in an increased phosphorylation of the alpha subunit of endogenous or exogenously added eIF-2, both in vitro and in vivo. These results suggest that a mechanism must exist in poliovirus-infected HeLa cells which prevents further phosphorylation of eIF-2 by the activated kinase. PMID- 3033311 TI - Expression of the Epstein-Barr virus gp350/220 gene in rodent and primate cells. AB - The gene encoding the Epstein-Barr virus envelope glycoproteins gp350 and gp220 was inserted downstream of the cytomegalovirus immediate-early, Moloney murine leukemia virus, mouse mammary tumor virus, or varicella-zoster virus gpI promoters in vectors containing selectable markers. Host cell and recombinant vector systems were defined which enabled the isolation of rodent or primate cell clones which expressed gp350/220 in substantial quantities. Continued expression of gp350/220 required maintenance of cells under positive selection for linked markers and periodic cloning. gp350/220 expressed in various host cells varied slightly in electrophoretic mobility, probably reflecting differences in glycosylation. Insertion of a stop codon into the gp350/220 open reading frame, upstream of the putative membrane anchor sequence, resulted in efficient secretion of truncated gp350 and gp220 from rat pituitary (GH3) cells. gp350/220 expressed in mammalian cells is highly immunogenic and elicits virus-neutralizing antibodies when administered to mice. PMID- 3033312 TI - Multiple integration site of hepatitis B virus DNA in hepatocellular carcinoma and chronic active hepatitis tissues from children. AB - Attention was directed to hepatitis B virus (HBV) integration in tissues obtained from an hepatocellular carcinoma (HCC) of an 11-year-old boy and from the liver of his 6-year-old brother, who had chronic active hepatitis. Multiple HBV DNA integration sites were demonstrated in both tissues. Cell population(s) in the HCC and liver from the patient with chronic active hepatitis were assumed to be heterogeneous with regard to HBV integration. The integrated forms in the two tissues showed similar genetic organization without gross rearrangement. The location of one of the virus-chromosomal junctions was restricted to the 5'-end region of the minus-strand DNA of HBV. The experimental results support our previous model for the mechanism of HBV integration, in which minus-strand replicative intermediates integrate into chromosomal DNA. The integrated HBV DNAs were conserved in the same region of the viral genome, spanning from the C gene through the S gene to the X gene, which contains intrinsic promoter-enhancer sequences. PMID- 3033313 TI - In vitro replication of mouse hepatitis virus strain A59. AB - An in vitro replication system for mouse hepatitis virus (MHV) strain A59 was developed using lysolecithin to produce cell extracts. In extracts of MHV infected cells, radiolabeled UMP was incorporated at a linear rate for up to 1 h into RNA, which hybridized to MHV-specific cDNA probes and migrated in denaturing formaldehyde-agarose gels to the same position as MHV genomic RNA. The incorporation of [32P]UMP into genome-sized RNA in vitro correlated with the observed increase of [3H]uridine incorporation in MHV-infected cells labeled in vivo. Incorporation of [32P]UMP into genome-sized RNA was inhibited when extracts were incubated with puromycin. The addition to the assay of antiserum to the MHV A59 nucleocapsid protein N inhibited synthesis of genome-sized RNA by 90% compared with the addition of preimmune serum. In contrast, antiserum to the E1 or E2 glycoproteins did not significantly inhibit RNA replication. In vitro synthesized RNA banded in cesium chloride gradients as a ribonucleoprotein complex with the characteristic density of MHV nucleocapsids isolated from virions. These experiments suggest that ongoing protein synthesis is necessary for replication of MHV genomic RNA and indicate that the N protein plays an important role in MHV replication. PMID- 3033314 TI - Cellular mutation mediates T-antigen-positive revertant cells resistant to simian virus 40 transformation but not to retransformation by polyomavirus and adenovirus type 2. AB - T-antigen-positive transformation revertant cell lines were isolated from fully simian virus 40 (SV40)-transformed Fisher rat embryo fibroblast cells (REF 52 cells) by methionine starvation. Reversion of the transformed cells (SV-52 cells) was caused by a mutation within the cellular genome. To demonstrate this, we isolated SV40 DNA from the host genome, inserted it into plasmid pSPT18 DNA, cloned it in Escherichia coli, and microinjected it into the nuclei of the REF 52 cells. Fully transformed cells were obtained with the same efficiency (20 to 25%) as after microinjection of wild-type SV40 DNA I. Furthermore, the revertant cells were resistant to retransformation by SV40. Following microinjection of wild-type SV40 DNA I, 42 independent cell lines were isolated. Cells of all analyzed lines acquired additional SV40 DNA copies, but changes in the cell morphology or growth characteristic were not demonstrable. However, the revertants were retransformable with a high efficiency after polyomavirus and adenovirus type 2 infections or microinjection. Also, fusion of the revertant cells with the grandparental REF 52 cells led to restoration of the transformed state. PMID- 3033316 TI - Binding of the pseudorabies virus immediate-early protein to single-stranded DNA. AB - In an attempt to correlate the ability to activate transcription with affinity for single-stranded DNA, both wild-type and temperature-sensitive pseudorabies virus immediate-early proteins were tested for the ability to bind to single stranded DNA columns. Wild-type and temperature-sensitive immediate-early proteins bound to nonspecific single-stranded DNA columns with similar affinities at both 0 and 40 degrees C. There did not seem to be a direct correlation between the ability to activate transcription and the ability to bind to single-stranded DNA. To study further the interactions that are involved in binding to single stranded DNA, we expressed the immediate-early protein in an Escherichia coli expression vector. In this system the expressed immediate-early protein was not phosphorylated, nor could it be complexed with mammalian cell factors. The first trp construct did not express a soluble form of the immediate-early protein, presumably due to the insoluble nature of the trp leader. We deleted a large segment of the trpE gene and found that the immediate-early fusion protein was soluble. We tested this protein for its affinity for single-stranded DNA by passage over single-stranded DNA cellulose columns. The bacterially expressed immediate-early protein bound single-stranded DNA at least as well as did the wild-type protein. Affinity for single-stranded DNA did not appear to be dependent on the phosphorylation state nor on the presence of mammalian cell factors. PMID- 3033315 TI - An aberrant avian leukosis virus provirus inserted downstream from the chicken c myc coding sequence in a bursal lymphoma results from intrachromosomal recombination between two proviruses and deletion of cellular DNA. AB - A chicken bursal lymphoma, LL6, contains avian leukosis virus DNA integrated 3' of the c-myc coding sequences, unlike all other examined bursal lymphomas, which have integrations 5' to c-myc. To better understand this unusual mutation, we examined a molecular clone containing the LL6 c-myc gene and determined the structure of the proviral insertion by DNA sequencing. Viral DNA begins 575 base pairs downstream of the c-myc coding sequences within the untranslated region, disrupting the use of the normal polyadenylation signal. An internal deletion of the provirus extends from within U3 in the 5' long terminal repeat to within the gp37-coding region of the env gene, disabling virus replication and protein synthesis. Both host-virus boundaries appear normal with respect to the site in viral DNA which is joined to host DNA; both long terminal repeats lack the terminal dinucleotide found in unintegrated DNA. However, in contrast to normal integrations, the six bases of cellular sequence at the 5' junction are not repeated at the 3' junction. The DNA sequences immediately downstream of the LL6 recombinant provirus are not part of the c-myc gene; they originate from the same chromosome as c-myc, but at least 15 kilobases (kb) away. In addition, DNA sequences normally residing 3' of c-myc are deleted in LL6. In summary, these results imply that the LL6 provirus is the result of recombination between two proviruses; that both proviruses were originally downstream of c-myc in the same orientation and separated by at least 15 kb; and that the recombination event was preceded, accompanied, or followed by an internal proviral deletion. No transcript could be detected within a 20-kb region downstream of the LL6 provirus, leaving unresolved the question of whether the additional chromosomal alterations make a specific contribution to LL6 tumorigenesis. PMID- 3033318 TI - Genetically determined resistance to lethal murine cytomegalovirus infection is mediated by interferon-dependent and -independent restriction of virus replication. AB - Susceptibility of 4-week-old mice of different strains to lethal murine cytomegalovirus (MCMV) infection was studied. Strains homozygous for H-2k and C57BL strains were resistant to greater than or equal to 10(5.5) PFU. B10.BR mice congenic for C57BL background genes and H-2k were about 10-fold more resistant than either C3H/HeN or C57BL strains. BALB/c mice (H-2d) were susceptible (50% lethal dose, 10(5.05) PFU). This susceptibility was dominant over resistance associated with H-2k but not that associated with C57BL background genes. The dominant susceptibility trait segregated in backcross mice as if carried by a single gene. Virus replication in spleen cells in vivo correlated with susceptibility to lethal infection. A similar trend was found in tests of salivary glands. Replication of MCMV in vitro in cultures of adherent spleen cells and primary mouse embryo cells correlated with replication in vivo. Neutralization of interferon (IFN) in cultures of adherent spleen cells reversed H-2k-linked restriction of viral replication but had minor effects on cells of other strains. Natural killer cell responses to infection were often higher in more resistant strains, but B10.BR mice developed minimal natural killer cell responses. Specific antibody and cytotoxic T cell responses in B10.BR mice were similar or lower than in other strains. Thus, resistance to lethal MCMV infection was not immunologically mediated, was dependent on and reflected by the capacity of cells from a given mouse strain to support replication in vivo and in vitro, and was IFN dependent and recessive if linked to H-2k but IFN independent when associated with C57BL background genes. PMID- 3033317 TI - Sequences responsible for erythroid and lymphoid leukemia in the long terminal repeats of Friend-mink cell focus-forming and Moloney murine leukemia viruses. AB - Despite the high degree of homology (91%) between the nucleotide sequences of the Friend-mink cell focus-forming (MCF) and the Moloney murine leukemia virus (MuLV) genomic long terminal repeats (LTRs), the pathogenicities determined by the LTR sequences of the two viruses are quite different. Friend-MCF MuLV is an erythroid leukemia virus, and Moloney MuLV is a lymphoid leukemia virus. To map the LTR sequences responsible for the different disease specificities, we constructed nine viruses with LTRs recombinant between the Friend-MCF and Moloney MuLVs. Analysis of the leukemia induced with the recombinant viruses showed that a 195 base-pair nucleotide sequence, including a 75-base-pair nucleotide Moloney enhancer, is responsible for the tissue-specific leukemogenicity of Moloney MuLV. However, not only the enhancer but also its downstream sequences appear to be necessary. The Moloney virus enhancer and its downstream sequence exerted a dominant effect over that of the Friend-MCF virus, but the enhancer sequence alone did not. The results that three of the nine recombinant viruses induced both erythroid and lymphoid leukemias supported the hypothesis that multiple viral genetic determinants control both the ability to cause leukemia and the type of leukemia induced. PMID- 3033319 TI - Class II polytropic murine leukemia viruses (MuLVs) of AKR/J mice: possible role in the generation of class I oncogenic polytropic MuLVs. AB - We examined the frequency of occurrence of polytropic murine leukemia viruses (MuLVs) in the spleens and thymuses of preleukemic AKR/J mice from 1 week to 6 months of age and analyzed the genomic RNAs of several polytropic isolates by RNase T1 oligonucleotide fingerprinting. Polytropic MuLVs were first detected in the spleens of 3-week-old mice and preceded the appearance of polytropic MuLVs in the thymus by over 1 month. At 4 months of age and older, nearly all mice expressed polytropic MuLVs in both organs. In contrast to previous studies which have identified class I polytropic MuLVs in AKR/J mice, fingerprint analysis of polytropic MuLVs from both young (3- to 4-week-old) and older (5- to 6-month-old) preleukemic mice indicated that a large proportion of viruses at both ages were class II polytropic MuLVs. All polytropic viruses (five isolates) analyzed from 3 to 4-week-old mice were recovered from spleen cells and were class II polytropic MuLVs. In older preleukemic mice, five of seven isolates were class II polytropic MuLVs and two were class I polytropic viruses. Class I and class II polytropic MuLVs were recovered from both the spleens and thymuses of older preleukemic mice. A detailed comparison of the class I and class II polytropic MuLVs from 5- to 6-month-old mice revealed that the nonecotropic gp70 sequences of most of the class I and class II MuLVs were identical, consistent with a common origin for these sequences. In contrast, the nonecotropic p15E sequences of class I MuLVs were clearly derived from different endogenous sequences than the nonecotropic p15E sequences of the class II MuLVs. The in vitro host ranges of class I and class II polytropic viruses were clearly distinguishable. Examination of the in vitro host range of several isolates suggested that the predominant polytropic viruses initially identified in the thymus (2 to 3 months of age) were class II polytropic viruses. The order of appearance of the class I and class II polytropic MuLVs and the identity of the gp70 oligonucleotides of these MuLVs suggested a model for the stepwise generation of class I polytropic MuLVs involving a class II polytropic MuLV intermediate. PMID- 3033321 TI - Sequence and structural organization of murine cytomegalovirus immediate-early gene 1. AB - In murine cytomegalovirus, abundant immediate-early transcription originates from 0.769 to 0.815 map units of the genome. This region contains the immediate-early gene (gene ieI) which encodes pp89, a phosphoprotein active in transcriptional regulation. In this paper we report on the precise location, structural organization, and sequence of gene ieI. The predominant ieI transcript, a 2.75 kilobase mRNA, is generated by splicing and composed of four exons. The precise termini of the 2.75-kilobase mRNA and the positions of the exons were determined by nuclease digestion experiments with either 5' or 3' end-labeled DNA fragments or in vitro transcribed cRNA probes. Exons of 300, 111, 191, and 1,703 nucleotides are separated by introns of 825, 95, and 122 nucleotides. The first AUG is located in the second exon of 111 nucleotides, and a single open reading frame of 1,785 nucleotides predicts a protein of 595 amino acids with a calculated molecular weight of 66,713. The N-terminal region of the protein contains sequences similar to a consensus sequence of histone 2B proteins. The regulatory function of pp89 and the role of this protein as an immunodominant antigen are discussed in relation to the amino acid sequence. PMID- 3033320 TI - Morphf mutants of Rous sarcoma virus: nucleotide sequencing analysis suggests that a class of morphf mutants was generated through splicing of a cryptic intron. AB - The nature of the lesions involved in producing the fusiform phenotype of three mutants (WO101, WO201, and tsST529) of the Schmidt-Ruppin A strain of Rous sarcoma virus (RSV) was determined by molecular cloning and DNA sequencing. WO101 and WO201 contained an in-frame deletion of the v-src region coding for amino acids 116 to 140 of p60v-src. The deleted segment was flanked by consensus splice donor and acceptor sequences and contained an appropriately positioned branchpoint acceptor consensus sequence, suggesting that the deletion occurred through an aberrant RNA splicing event. S1 mapping experiments performed on RNA isolated from chicken cells infected with molecularly cloned wild-type RSV DNA suggested that the splice acceptor involved in the generation of this deletion was utilized at a low frequency (less than 1.0%) in wild-type RSV-infected cells. These results suggested that stable mutations may have arisen in the coding sequence of a eucaryotic viral transforming gene as a result of a probable aberrant RNA splicing event followed by reverse transcription into DNA. ST529 was found to harbor the same deletion present in WO101 and WO201 but also contained a point mutation which resulted in the substitution of lysine for glutamic acid at position 93. This change and the resulting large change in local charge were presumably required for the temperature-sensitive transformation phenotype of ST529. These results, together with other known deletions that produce fusiform mutants, suggested that a region within the amino-terminal one-third coding region of the src gene contributed to a structural domain of p60v-src that was important for controlling some morphological parameters of transformation in cells infected with RSV. PMID- 3033322 TI - Retention or loss of v-mil sequences after propagation of MH2 virus in vivo or in vitro. AB - During propagation of the defective avian retrovirus MH2 in the presence of replication-competent helper virus, deletion of portions of the viral genome occurred frequently. After transformation of quail cells in vitro, v-mil sequences were lost, leading to populations of MH2 viruses which were highly deficient for mil gene expression but which could transform macrophage and fibroblast cells in vitro with high efficiency. In contrast, after induction of tumors in quail with mil-deficient MH2 viral stocks, a majority of the tumor DNAs contained mil+ proviruses, suggesting that there is selection for retention of the v-mil gene in vivo and that the mil protein may play a role in the oncogenicity of MH2 virus. We also isolated MH2-transformed cell lines which contained deleted proviruses arising from packaging and subsequent integration of the subgenomic v-myc-encoding mRNA. Some of these cell lines produced viruses which encoded abnormal v-myc proteins and had altered in vitro transforming properties. These altered phenotypes may be caused by mutations within the v-myc gene. PMID- 3033323 TI - Efficient resolution of replicated poxvirus telomeres to native hairpin structures requires two inverted symmetrical copies of a core target DNA sequence. AB - The terminal hairpin sequences of the linear double-stranded DNA genome of the leporipoxvirus Shope fibroma virus (SFV) has been cloned in Saccharomyces cerevisiae and in recombination-deficient Escherichia coli as a palindromic insert within circular plasmid vectors. This sequence configuration is equivalent to the inverted repeat structure detected as a telomeric replicative intermediate during poxvirus replication in vivo. Previously, it has been shown that when circular plasmids containing this palindromic insert were transfected into SFV infected cells, efficient replication and resolution generated linear minichromosomes with bona fide viral hairpin termini (A. M. DeLange, M. Reddy, D. Scraba, C. Upton, and G. McFadden, J. Virol. 59:249-259, 1986). To localize the minimal target DNA sequence required for efficient resolution, a series of staggered unidirectional deletions were constructed at both ends of the inverted repeat. Analyses of the resolution efficiencies of the various clones indicate that up to 240 base pairs (bp) centered at the symmetry axis were required for maximal resolution to minichromosomes. To investigate the role of the AT-rich central axis sequences, which in SFV include 8 nonpalindromic bp, a unique AflII site at the symmetry axis was exploited. Bidirectional deletions extending from this AflII site and insertions of synthetic oligonucleotides into one of the deletion derivatives were constructed and tested in vivo. The efficiency with which these plasmids resolved to linear minichromosomes with hairpin termini has enabled us to define the minimal target DNA sequence as two inverted copies of an identical DNA sequence between 58 and 76 bp in length. The nonpalindromic nucleotides, which, after resolution, constitute the extrahelical residues characteristic of native poxviral telomeres, were not required for resolution. The close resemblance of the SFV core target sequence to the analogous region from the orthopoxvirus vaccinia virus is consistent with a conserved mechanism for poxviral telomere resolution. PMID- 3033324 TI - Genetic and biological analyses of a herpes simplex virus intertypic recombinant reduced specifically for neurovirulence. AB - RS6 is a herpes simplex virus intertypic recombinant derived from type 1 strain 17 syn+ and type 2 strain HG52. With a 50% lethal dose of about 10(5) PFU after intracerebral inoculation of mice, RS6 was approximately 100,000 times less neurovirulent than either of its wild-type parental viruses were. When compared with strains 17 syn+ and HG52, RS6 replicated intermediately in primary mouse embryo fibroblasts in vitro at 38.5 degrees C (mouse temperature) and to wild type peak titers in mouse feet in vivo. In contrast, following intracranial inoculation of mice, RS6 replicated significantly less well than did either of its parental viruses in brains. The genetic defect(s) responsible for the reduced neurovirulence of RS6 was stable after in vitro and in vivo serial passage, was not manifested as temperature-sensitive plaquing in vitro, and did not affect thymidine kinase expression. These data indicate that RS6 has a genetic defect(s) specifically affecting its ability to replicate in the mouse brain. Using marker rescue technologies, we increased the neurovirulence of RS6 and localized one genetic determinant(s) involved with the reduced neurovirulence of this agent to 0.72 to 0.87 map units (and, tentatively, to 0.79 to 0.83 map units) of the herpes simplex virus genome. When coupled with the work suggesting that thymidine kinase expression is essential for efficient replication in nerve tissues and earlier reports from this laboratory and others, the results presented in this study indicate that more than one herpes simplex virus gene is involved with neurovirulence. PMID- 3033325 TI - Two strains of Epstein-Barr virus (B95-8 and a P3HR-1 subclone) that lack defective genomes induce early antigen and cause abortive infection of Raji cells. AB - The heterogeneity of Epstein-Barr virus (EBV) obtained from P3HR-1 cells has permitted derivation of a distinct subclone of P3HR-1 (L. Heston, M. Rabson, N. Brown, and G. Miller, Nature (London) 295:160-163, 1982). We have analyzed the biologic properties and genomic structure of this subclonal virus (clone 13) compared with those of parental P3HR-1 and B95-8 viruses. Synthesis of EBV compared with those of parental P3HR-1 and B95-8 viruses. Synthesis of EBV proteins in Raji cells superinfected with virus derived from P3HR-1, clone 13, and B95-8 was analyzed both by fluorography of radiolabeled proteins and by immunoblotting. Highly concentrated preparations of clone 13 and B95-8 virus induced most of the spectrum of EBV proteins in Raji cells with the exception of the 145,000-, 140,000-, and 110,000-molecular-weight proteins, which were either undetectable or reduced. Moreover, both clone 13 and B95-8 viruses also induced the same patterns of early antigen diffuse components as the parental P3HR-1 virus did. However, only P3HR-1 virus could induce EBV DNA synthesis in superinfected Raji cells, as determined both by buoyant density centrifugation and by in situ cytohybridization with biotinylated recombinant EBV DNA probes. Defective heterogeneous molecules present in P3HR-1 virus have been implicated in early antigen induction after superinfection of Raji cells. Therefore, Southern blots of clone 13, P3HR-1, and B95-8 viruses were hybridized to recombinant EBV fragments representing the sequences contained within the defective molecules in P3HR-1. The parental P3HR-1 contained the previously described defective molecules. No evidence for defective molecules was found in clone 13 or B95-8 viruses. These data indicate that concentrated preparations of both clone 13 and B95-8 viruses can induce abortive infection in Raji cells, but while the defective molecules are not needed for induction of early antigen diffuse components, they may be required for the induction of viral DNA synthesis. PMID- 3033326 TI - Possible mechanisms by which the H-2Kbm3 mutation may decrease cytotoxic T lymphocyte recognition of vesicular stomatitis virus nucleoprotein antigen. AB - Spleen cells from C57BL/6 (B6) mice generate a strong in vitro cytotoxic T lymphocyte (CTL) response specific for vesicular stomatitis virus (VSV). Spleen cells from VSV-primed B6-H-2bm3 (bm3) mice, which have a mutation in H-2Kb, require approximately 10-fold more UV-inactivated VSV to generate in vitro secondary anti-VSV CTL, compared with spleen cells from primed B6 mice. Anti-VSV CTL elicited in both bm3 and B6 mice are primarily specific for the viral nucleocapsid protein (N protein), as demonstrated by using recombinant vaccinia viruses that express the VSV N protein. bm3 CTL were found to exhibit only a very low level of lytic activity when tested against autologous VSV-infected concanavalin A spleen cell blasts as well as several H-2b tumor cell lines. The weak anti-VSV response of bm3 CTL was found to be the result of a combination of inefficient recognition of VSV-infected target cells and decreased elicitation of secondary effector cells. VSV-infected bm3 target cells were not killed as well as B6 targets by either bm3 or B6 effectors. This is because of the inefficient recognition of targets, as demonstrated by the fact that VSV-infected bm3 cells were unable to competitively inhibit the lysis of VSV-infected B6 target cells by either bm3 or B6 effectors. By using cells from recombinant mice, it was shown that the CTL response restricted by H-2Kb was low in the bm3 mice, compared with that of the B6 mice. However, the H-2Db-restricted CTL activity was similarly low in both the B6 and bm3 mice. The possibility that the low response to VSV infected bm3 cells is caused by differences between the bm3 and B6 cells in expression of either viral antigens or H-2K was investigated by radiolabeling and immunoprecipitation. VSV-infected B6 and bm3 cells were found to express equivalent levels of both viral antigens and H-2K. These results indicate that the bm3 mutation alters a functional site on the H-2Kb molecule that is involved in the recognition of VSV-infected cells. The observation that elicitation of bm3 CTL can occur at high antigen doses further suggests that the bm3 mutation results in a lower affinity of H-2K either for viral antigen or for receptor sites on the CTL. PMID- 3033327 TI - Avian sarcoma and leukosis virus pol-endonuclease recognition of the tandem long terminal repeat junction: minimum site required for cleavage is also required for viral growth. AB - Integration of retroviral DNA is a site-specific reaction involving an endonuclease encoded by the viral pol gene (pol-endo). In vitro the pol-endo from avian sarcoma and leukosis viruses (ASLVs) cleaves both DNA strands near the U5 U3 junction of tandem long terminal repeats (LTR-LTR junction) in single-stranded and replicative form (RF)-I substrates. We have reported previously that the sequences that are required for cleavage of single-stranded substrates by the alpha beta form of the pol-endo differ for the plus and minus strands (G. Duyk, M. Longiaru, D. Cobrinik, R. Kowal, P. deHaseth, A. M. Skalka, and J. Leis, J. Virol. 56:589-599, 1985). This is not the case with RF-I substrates, in which a maximum of 22 base pairs of U5 and 8 base pairs of U3 were required for alpha beta pol-endo cleavage in each strand. Insertion of a palindromic octanucleotide (CATCGATG) at the LTR-LTR junction abolished cleavage in RF-I but not in single stranded DNA substrates. Deletion of the four nucleotides (TTAA) at the junction prevented cleavage in the plus strand of RF-I DNA, but did not affect cleavage of single-stranded DNA. Furthermore, the alpha beta form of ASLV pol-endo did not recognize heterologous LTR-LTR junction sequences from the reticuloendotheliosis virus or Moloney murine leukemia virus in either substrate form, despite their sequence and structural similarities to the ASLV junction. These results support a role for a sequence-specific interaction between the ASLV pol-endo and the LTR LTR junction domains that are required for cleavage. By using the infectious Rous sarcoma virus clone pATV8-K, we introduced a set of deletions into the U5 region that would be incorporated into the LTR-LTR junction on viral replication. In the unintegrated provirus, the deletions started 43 base pairs from the LTR-LTR junction and extended various lengths toward the junction. Results of transfection studies with these clones indicated that the U5 sequences that are required for virus production in vivo correspond to those that are required for cleavage of RF-I DNA in vitro. PMID- 3033328 TI - Characteristics of the specific cell-mediated immune response in human immunodeficiency virus infection. AB - The human immunodeficiency virus (HIV)-specific lymphocyte proliferation response was determined for 40 persons at different stages of HIV infection. The specific response to purified HIV virion antigens from strain HTLV-IIIB was poor, occurred in only 9 of the 40 subjects, was not improved with the addition of interleukin 2, and was more frequent in symptom-free individuals (46%) than in patients with lymphadenopathy syndrome (10%). Reactivity to subcomponent p24 was better than that to whole HIV; reactivity was present in five of six infected persons and increased with the addition of exogenous interleukin-2. Reactivities to subcomponents (g)p41 and gp120 were also measured. This is the first evidence of a specific cell-mediated immune response to HIV antigen in HIV-infected persons. Monkeys immunized with purified HIV or with purified p24 displayed cellular immunoreactivity both to whole HIV and to subcomponents. In contrast to the poor reactivity to HIV antigen, the lymphocytes of the patients had good specific cell proliferation responses to cytomegalovirus and herpes simplex virus challenge and a normal response to the addition of phytohemagglutinin. The results suggest a functional defect in peripheral lymphocytes of some HIV-infected individuals on the basis of their response to whole HIV antigen and a better response to gag protein. PMID- 3033329 TI - Relationship between simian virus 40 large tumor antigen expression and tumor formation in transgenic mice. AB - A line of transgenic mice containing the simian virus 40 (SV40) large tumor antigen gene under the control of the viral enhancer-promoter expressed this viral protein in the brains of these mice within the first 2 weeks after birth. Multiple foci of anaplastic cells formed in the choroid plexuses of these mice at 36 to 41 days after birth, and normal tissue coexisted with these transformed foci. Immunoperoxidase staining to detect the SV40 T antigen showed tumor specific expression of nuclear T antigen at late times in tumor development, approximately 90 to 100 days and thereafter. The level of SV40 T antigen, on a per cell basis, appeared to be lower in the great majority of choroid plexus cells at earlier times in tumor development. These results suggest that low levels of tumor antigen (14 to 36 days) are present before detectable pathology (36 to 41 days) and the level of T antigen per cell is higher in rapidly growing late-stage tumors (older than 90 days). PMID- 3033330 TI - Effects of cDNA hybridization on translation of encephalomyocarditis virus RNA. AB - Cell-free translation of the RNA of encephalomyocarditis virus was examined after hybridization of chemically synthesized cDNA fragments to different sites of the 5' noncoding region of the viral RNA. The following results were obtained. The binding of cDNA fragments to the first 41 nucleotides, to the poly(C) tract (between nucleotides 149 and 263), and to the sequence between nucleotides 309 and 338 did not affect translation of the viral RNA; the binding of cDNA fragments to the sequence between nucleotides 420 and 449 caused a slight inhibition; and the binding of fragments to eight different sites between nucleotides 450 and the initiator AUG codon (nucleotide 834) caused high degrees of inhibition. The results suggest that the first part of the 5' untranslated region, at least to nucleotide 338, may not be required for encephalomyocarditis viral RNA translation; however, the region near nucleotide 450 is important for translation of the viral RNA. The possibility that initiation occurs at an internal site is discussed. PMID- 3033331 TI - Coronavirus E1 glycoprotein expressed from cloned cDNA localizes in the Golgi region. AB - Cloned cDNA encoding the membrane glycoprotein E1 of the coronavirus mouse hepatitis virus strain A59 was expressed transiently in a monkey fibroblast cell line (COS) by using a simian virus 40-based vector. As determined by indirect immunofluorescence microscopy, the E1 protein accumulated intracellularly in a perinuclear region coincident with a Golgi marker. The same three species of E1 that occur in virus-infected cells were also found in transfected cells. These are one unglycosylated form and two apparently O-glycosylated forms that could be labeled in a tunicamycin-resistant fashion with [3H]glucosamine. Because O glycosylation occurs posttranslationally in the Golgi apparatus, we could show, by monitoring the rate of acquisition of oligosaccharides, that the transport of E1 from the rough endoplasmic reticulum to the Golgi apparatus had a half time of between 15 and 30 min. PMID- 3033332 TI - Identification of viral polypeptides involved in pseudorabies virus ribonucleotide reductase activity. AB - We studied pseudorabies virus-induced ribonucleotide reductase and found that it exhibited biochemical properties very similar to those of herpes simplex virus reductase. A polyclonal rabbit antiserum (P9) directed against the carboxy terminus of subunit H2 polypeptide (38,000 daltons) of herpes simplex virus reductase neutralized the pseudorabies virus reductase, as well as the herpes simplex virus isozyme. This serum recognized two pseudorabies virus-specified polypeptides of 34,000 and 110,000 daltons, which may represent the two subunits of the enzyme. Furthermore, as already shown for herpes simplex virus reductase (E. A. Cohen, P. Gaudreau, P. Brazeau, and Y. Langelier, Nature [London] 321:441 443, 1986), we show that the nonapeptide itself specifically inhibited pseudorabies reductase activity. PMID- 3033333 TI - Cell-protective monoclonal antibodies to bovine enterovirus-3 and partial or no activity against other serotypes. AB - Preparation of monoclonal antibodies to bovine virus diarrhea virus (BVDV) yielded some hybridoma cells that secreted monoclonal antibodies against the Madin-Darby bovine kidney cells. The anti-cellular monoclonal antibodies reacted with other bovine cells (bovine turbinate and testicle) but not with cell lines derived from other animal species. Subclones derived from one hybridoma partially blocked the infectivity of BVDV, possibly through the binding of the monoclonal antibodies with an epitope close to the receptor site of BVDV and not by way of steric hindrance. Unexpectedly, these same subclones completely blocked the infectivity of bovine enterovirus-3 (BEV-3) strain 240A and partially blocked the infectivity of BEV-2 and BEV-3 (ATCC strain) but not that of other serotypes. Other subclones derived from two other hybridomas, although cell membrane specific, did not have a protective activity against BEV or BVDV. PMID- 3033335 TI - Nephroblastoma in an ovotestis of a true hermaphrodite: a case report. AB - We describe a patient who presented with a painful mass in the left inguinoscrotal region, gynecomastia, perineal hypospadias with chordee and a bifid scrotum with a small gonad in the right hemiscrotum. At operation the left mass was found to be an ovotestis with hemorrhage in a corpus luteum cyst. A rudimentary uterus and fallopian tube were situated next to the ovotestis. The right gonad also was an ovotestis with a tiny focus of small cell malignancy that was suggestive of nephroblastoma. PMID- 3033334 TI - Host immune response to cytomegalovirus: products of transfected viral immediate early genes are recognized by cloned cytolytic T lymphocytes. AB - To confirm that immediate-early (IE) genes of murine cytomegalovirus (MCMV) give rise to antigens recognized by specific cytolytic T lymphocytes (CTL), a 10.8 kilobase fragment of MCMV DNA which is abundantly transcribed at IE times was transfected into L cells expressing the Ld class I major histocompatibility glycoprotein. The viral genome fragment contains sequences of the three IE transcription units of MCMV: ie1, ie2, and ie3. In the transfected cell lines, only the predominant 2.75-kilobase transcript of ie1 and its translation product pp89 could be detected. The transfectants were analyzed for membrane expression of an IE antigen by employing clone IE1, an IE-specific CTL clone, as the probe. Only cells that expressed both the MCMV IE gene(s) and the Ld gene were recognized by the CTL clone. PMID- 3033336 TI - Extragonadal germ cell tumor: the preoperative urological evaluation. AB - Primary extragonadal germ cell tumors were diagnosed in 4 patients in whom surgical or postmortem examination of the testis failed to reveal germ cell tumor. A fifth patient underwent serial examination with testicular ultrasound without evidence of any abnormalities and he remains free of disease following chemotherapy. Based on our experience and a literature review, it is our opinion that orchiectomy is not indicated unless there is a palpable testicular abnormality, recent change in testicular size or consistency, abnormal testicular ultrasound or history of cryptorchidism, or if the primary tumor is choriocarcinoma. PMID- 3033337 TI - A cluster of patients with a chronic mononucleosis-like syndrome. Is Epstein-Barr virus the cause? AB - A cluster of 134 patients who had undergone Epstein-Barr virus (EBV) serological testing because of suspected chronic EBV syndrome was investigated in Nevada. Fifteen case-patients were identified who had severe, persistent fatigue of undetermined etiology for more than two months. When compared with the remaining 119 patients who had less severe illnesses and with 30 age-, sex-, and race matched control-persons, these 15 patients had significantly higher antibody titers against various components of EBV and against cytomegalovirus and herpes simplex and measles viruses. Epstein-Barr virus serology could not reliably differentiate individual case-patients from the others, and the reproducibility of the tests within and among laboratories was poor. As a group, the case patients appear to have had a syndrome that is characterized by chronic fatigue, fever, sore throat, and lymphadenopathy. The relationship of this fatigue syndrome to EBV is unclear; further studies are needed to determine its etiology. PMID- 3033338 TI - Frequency of 'chronic active Epstein-Barr virus infection' in a general medical practice. AB - Twenty-one percent of 500 unselected patients, aged 17 to 50 years, seeking primary care for any reason were found to be suffering from a chronic fatigue syndrome consistent with "chronic active Epstein-Barr virus (EBV) infection," They had been experiencing "severe" fatigue, usually cyclic, for a median of 16 months (range, six to 458 months), associated with sore throat, myalgias, or headaches; 45% of the patients were periodically bedridden; and 25% to 73% reported recurrent cervical adenopathy, paresthesias, arthralgias, and difficulty in concentrating or sleeping. The patients had no recognized chronic "physical" illness and were not receiving psychiatric care. While antibody titers to several EBV-specific antigens were higher in patients than in age- and sex-matched controls subjects, the differences generally were not statistically significant. A chronic fatigue syndrome consistent with the chronic active EBV infection syndrome was prevalent in our primary care practice. However, our data offer no evidence that EBV is causally related to the syndrome. Indeed, we feel that among unselected patients seen in a general medical practice currently available EBV serologic test results must be interpreted with great caution. PMID- 3033340 TI - EB or not EB--that is the question. PMID- 3033339 TI - Effect of the contraceptive sponge on chlamydial infection, gonorrhea, and candidiasis. A comparative clinical trial. AB - To investigate the effect of the nonoxynol 9-impregnated contraceptive sponge on the incidence of chlamydial infection, gonorrhea, and candidiasis, we conducted a randomized comparative study among high-risk women in Bangkok, Thailand. The first (parallel) portion of the study covered 434 woman-weeks among sponge users and 494 woman-weeks among nonusers. As compared with women not using the sponge, sponge users were found to be less likely to become infected with chlamydia (relative rate, 0.67; 95% confidence interval, 0.42 to 1.07) and gonorrhea (relative rate, 0.31 [0.16 to 0.60]) but more likely to become infected with Candida (relative rate, 2.76 [0.96 to 7.98]). Women who continued in the study were crossed over to the alternate group, with former nonusers starting to employ the sponge and vice versa. The results of this second phase were similar to those of the larger parallel study. Overall, these results suggest that women using the sponge are protected against the two most common sexually transmitted pathogens, which are also those with the most serious health consequences. However, women using the sponge should be advised they may have an increased likelihood of a vaginal infection with Candida. PMID- 3033341 TI - Cervical dysplasia upturn worries gynecologists, health officials. PMID- 3033342 TI - For women infected with papillomavirus, close watch counseled. PMID- 3033343 TI - Hepatoblastoma in families with polyposis coli. AB - We observed hepatoblastoma in four unrelated children who have a family history of polyposis coli and found this association in ten other kindreds in the literature. The one child in our series who has survived hepatoblastoma showed multiple colonic adenomas at 7 years of age. She and eight affected maternal relatives also had congenital hypertrophy of the retinal pigment epithelium, a new marker of gene carriers in some polyposis coli families. These findings suggest that childhood hepatoblastoma is an associated feature of familial polyposis coli. PMID- 3033344 TI - Acute haemorrhagic conjunctivitis. PMID- 3033345 TI - Abnormality of circulatory reflex and aldosterone response during head-up tilting in patients with primary aldosteronism. AB - Abnormality of the circulatory reflexes has been reported in patients with primary aldosteronism. However, changes in blood pressure, heart rate, plasma renin activity (PRA), and plasma aldosterone concentration (PAC) after head-up tilting in primary aldosteronism have not yet been reported. Seven patients with primary aldosteronism were tilted to a 65 degree head-up position which was maintained for 30 min. Systolic blood pressure decreased significantly 5 min after tilting and remained at this level during the period of tilting. Diastolic blood pressure did not change during the tilting. Heart rate increased after 5 min of tilting and this level of heart rate was maintained for 30 min. Plasma renin activity was low and did not change during tilting. However, plasma aldosterone concentration increased significantly 20 min after tilting. Plasma cortisol concentration and plasma ACTH concentration also increased significantly. These results suggest that primary aldosteronism causes abnormalities of the circulatory reflexes. The increase of endogenous ACTH may increase plasma aldosterone concentration in patients with primary aldosteronism. PMID- 3033346 TI - The mechanism of the control of renin release by beta-adrenergic receptors. AB - Recent reports suggest that prostaglandins play an important role in the beta adrenergic receptor mechanism of renin release. However, the site of the action of prostaglandins has not yet been clarified. Superfusion of rabbit renal cortical slices was used to evaluate the beta-adrenergic receptor mechanism of renin release. Renin release was stimulated by isoproterenol, prostaglandin E2, and dibutyryl cyclic AMP. Renin release stimulated by isoproterenol was inhibited by propranolol, whereas renin release stimulated by prostaglandin E2 was not inhibited by propranolol. Isoproterenol stimulated prostaglandin E2 release as well as renin release, and indomethacin inhibited these effects of isoproterenol. Propranolol inhibited prostaglandin E2 release stimulated by isoproterenol. On the other hand, indomethacin did not affect renin release stimulated by prostaglandin E2 release. Dibutyryl cyclic AMP did not stimulate prostaglandin E2 release. Indomethacin did not affect renin release stimulated by dibutyryl cyclic AMP, however, it suppressed prostaglandin E2 release during the superfusion with dibutyryl cyclic AMP. Finally, isoproterenol and prostaglandin E2 stimulated cyclic AMP release. These data suggest that prostaglandins play an important role in the beta-adrenergic receptor mechanism of renin release and the site of the action of prostaglandins is between the beta-adrenergic receptor and cyclic AMP. PMID- 3033347 TI - [The use of CT in the treatment of lung cancer with radiation therapy]. AB - In this study, the extent to which CT accuracy in the diagnosis of regional lymph node metastasis of non-small cell lung cancer contributed to treatment results with radiation therapy was examined. The long-termed survival rate of 133 cases examined by CT scanning was superior to that of 154 cases diagnosed by conventional radiological method alone. The difference was statistically significant in N2 cases. When the mediastinal lymph node metastasis was detected on the ipsilateral side of the primary tumor or when the mediastinal lymph node swelling was smaller than 3 cm in diameter, the prognosis became better. PMID- 3033349 TI - [Partition of the motor cortex and its output--f. The occipital-temporal cortex and voluntary movement]. PMID- 3033348 TI - [A case of malignant paraganglioma arising from the retroperitoneum]. AB - We report the case of 38-year-old male with a malignant and functional paraganglioma arising from the retroperitoneum. He was admitted to our hospital with a pathological fracture of the right femur, A diagnosis of paraganglioma was made after taking a biopsy specimen and discovering a hypervascular retroperitoneal tumor. The level of catecholamine in his urine was greatly increased, accompanied with hypertension, rapid pulse and sweating. He was treated with chemotherapy, radiotherapy and surgical therapy. However, he died from pulmonary metastasis. PMID- 3033350 TI - [Theory of the neural circuit network of memory and recognition]. PMID- 3033351 TI - [Molecular neurobiology of memory--history and present status]. PMID- 3033352 TI - [A case of spontaneous gas formation in hepatocellular carcinoma (HCC)]. PMID- 3033353 TI - [Two cases of skeletal metastasis of hepatocellular carcinoma demonstrating an abnormal accumulation of Tc-99m PMT]. PMID- 3033354 TI - [High resolution CT of the lung after lymphangiography]. PMID- 3033355 TI - [Treatment results in lung cancer using radiotherapy]. PMID- 3033356 TI - [Ultrasonography of peripheral cholangiocellular carcinoma--comparison with pathologic findings in 7 cases]. PMID- 3033357 TI - [Intralesional interferon in the treatment of extramammary Paget's disease]. PMID- 3033358 TI - [Studies on production of human epidermal growth factor and its receptor in human hepatoma cell line (PLC/PRF/5)]. PMID- 3033359 TI - [Pathophysiological significance of trace metals in rats with experimental liver cirrhosis]. PMID- 3033360 TI - [Biochemical studies of inhibition by interferon (alpha + beta) in mouse liver regeneration after partial hepatectomy]. PMID- 3033361 TI - [The development of new tumor imaging 99mTc (V)-DMS kit: preparation and labeling condition studies]. PMID- 3033362 TI - A case of rhabdomyolysis in chronic renal failure. AB - Rhabdomyolysis can be induced by a variety of physical and chemical insults to skeletal muscle. Though there are only a few reports of myositis associated with viral diseases, and no reported cases of virus-induced rhabdomyolysis in uremic patients. Chronic uremic states have been known to potentiate a variety of metabolic and immunological abnormalities. Present case had an acute, progressive and fatal rhabdomyolysis which was thought to be induced by virus infection. A 43 year-old man had received hemodialysis therapy for 8 years. He suffered from an upper respiratory tract infection 11 days before admission. On admission, he was diagnosed as rhabdomyolysis with severe lactic acidosis and marked hyperkalemia. Although intensive care had been performed, he died of uncontrollable hyperkalemia (10.0 mEq/L) 2 days after admission. Maximum CPK and GOT levels were 105,200 and 56,800 mU/ml, respectively. Most probable cause of rhabdomyolysis was thought to be Parainfluenza type-3 infection, though histological examination failed to prove virus infection. PMID- 3033363 TI - Effect of peptide bond splitting on ouabain sensitive conformational changes in Na+,K+-ATPase treated with N-[p-(2-benzimidazolyl)phenyl]maleimide. AB - Trypsin treatment of N-[p-(2-benzimidazolyl)phenyl]maleimide modified enzyme caused a marked reduction in Na+,K+-ATPase activity and in the amount of the alpha-chain, which contains the phosphorylation and ouabain binding sites. However, these preparations retained nearly 90% of the ouabain binding capacity and showed ouabain sensitive dynamic fluorescence changes accompanying the hydrolysis of ATP. The data showed that the three dimensional structure of Na+,K+ ATPase, which is important in the dynamic fluorescence change, is little affected in spite of extensive covalent bond splitting in the alpha-chain of Na+,K+ ATPase. PMID- 3033364 TI - Behavioral and neurochemical changes produced by postnatal pretreatments with methamphetamine in rats. AB - Male neonates of Wistar strain rats were given s.c. 1-4 mg/kg/day of methamphetamine (MAP) for 7 successive days from days 6 to 12 after birth. The acquisition processes of the discriminative lever-press avoidance response were investigated from day 60 after birth. Effects of the postnatal pretreatments with MAP on saturation constants for specific bindings of 3H-spiperone (SPP) and 3H WB4101, respectively, in 8 brain regions were also investigated at 100-120 days after birth. In addition, dopamine, noradrenaline and the levels of their metabolites were measured in the brain. No significant difference was detected in body weight, gross behaviors and avoidance learning between saline- and MAP pretreated groups. However, effects of MAP and apomorphine on the locomotor activity significantly increased in the MAP-pretreated group. Significant decreases in Bmax and Kd values of 3H-SPP binding sites in the striatum were detected in the MAP-pretreated group, while significant decreases in Bmax values of 3H-WB4101 binding sites in the cortex and hippocampus as well as those in Kd values in the hippocampus were found in the treated group. Dopamine and noradrenaline levels significantly decreased in the MAP-pretreated group, but on the contrary, their metabolites levels significantly increased. These results suggest that postnatal pretreatments with MAP produce long-lasting behavioral changes associated with an impaired development of catecholaminergic neurons in the rat brain after maturity. PMID- 3033365 TI - An increase of tissue cyclic AMP level by adenosine in the dog pancreas without stimulation of exocrine secretion. AB - In the vascularly isolated and self-hemoperfused dog pancreas, secretin (0.025 clinical units) and adenosine (1.0 mg) administered intra-arterially (i.a.) increased tissue cyclic AMP level by about 46% and 37%, respectively. Although the increases in the nucleotide were not statistically different, only secretin stimulated exocrine secretion. The increase in cyclic AMP induced by adenosine was significantly reversed by pretreatment with theophylline (0.3 mg, i.a.). These results suggest that the effect of adenosine on cyclic AMP formation occurs mainly in the non-exocrine system of the dog pancreas through A2/Ra-receptors. PMID- 3033366 TI - Peripheral transmission in primary sensory nerves. AB - Primary sensory nerves do not only transfer messages to the central nervous system but also transmit messages to the periphery. The peripheral response is one of defense, including neurogenic inflammation such as miosis and a breakdown of the blood aqueous barrier in the eye or vasodilatation and plasma extravasation in the skin. Recent pharmacological, biochemical and immunohistochemical studies have revealed that tachykinins are the most likely transmitters at peripheral as well as central endings of primary sensory nerves. Furthermore, studies with trigeminal nerve-innervated and isolated rabbit iris sphincter muscle have clearly shown that the peripheral neurotransmission is affected by many neuromodulators and irritants. It is not unlikely that the neurotransmission mechanisms at peripheral endings are analogous to those at central endings in the primary sensory nerves. PMID- 3033367 TI - Effect of concanavalin A on 5'-nucleotidase activity of rabbit blood platelets. AB - Ecto-5'-nucleotidase (ecto-5'-NU) of platelets was enhanced by concanavalin A (Con A). This effect of Con A was antagonized by alpha-methyl-D-mannose, a specific antagonist of Con A binding to glycoprotein. Coformycin, an adenosine deaminase inhibitor, did not change the effect of Con A on the ecto-5'-NU. Uptake of adenosine by platelets was not affected by Con A. It was suggested that the ecto-5'-NU of platelet might be a direct and primary site of action of Con A. PMID- 3033369 TI - Treatment of epilepsy in infancy with special emphasis on ACTH therapy. PMID- 3033368 TI - Inhibitory effects of condensed tannins on angiotensin converting enzyme. AB - Effects of condensed tannins isolated from Rhei Rhizoma on the activities of angiotensin converting enzyme (ACE) and various proteases were examined in vitro. Among the various condensed tannins tested, procyanidin B-5 3,3'-di-O-gallate and procyanidin C-1 3,3',3"-tri-O-gallate strongly inhibited the activity of ACE. The concentration of procyanidin B-5 3,3'-di-O-gallate required for 50% inhibition of ACE was 1.3 X 10(-6) M. The inhibition of ACE by condensed tannins was reversible and non-competitive, according to dialysis and to Dixon plots. However, over one hundred times the concentration was required to inhibit activities of other proteases such as trypsin, chymotrypsin, leucine aminopeptidase, carboxypeptidase A and urinary kallikrein. These results suggest that the inhibitory effects of condensed tannins on the activities of ACE are specific. PMID- 3033370 TI - Change in brain thyrotropin-releasing hormone (TRH) mechanism of amygdaloid kindled rats. AB - We reported previously that DN-1417, a potent analog of thyrotropin-releasing hormone (TRH), suppressed both the progression of amygdaloid (AM) kindling and AM kindled seizure. To study a functional role of the cerebral TRH mechanism in AM kindling, immunoreactive TRH (IR-TRH) and specific TRH receptor binding were examined in the rat brains kindled from the left AM. The IR-TRH concentration elevated significantly in the amygdala plus piriform cortex and the hippocampus 24 and 48 hours after the AM kindled convulsion. Such an elevation of IR-TRH was not found 7 days after the last convulsion, indicating that the elevation of IR TRH was a transient change seen after the AM kindled convulsion. By contrast, the specific TRH receptor binding in the striatum increased 48 hours, 7 and 21 days after the AM kindled convulsion. This indicates that the increase of the specific TRH binding in the striatum was a long-lasting change. The present study suggests that the change in the striatal TRH receptors may be associated with a long lasting seizure susceptibility of AM kindled rats. PMID- 3033371 TI - The possible involvement of free radicals in seizure mechanism. PMID- 3033372 TI - Role of cyclic AMP and intracellular calcium concentration in seizure activity. PMID- 3033373 TI - Correlation between inositide response and seizure-related substances as well as antiepileptic drugs. PMID- 3033375 TI - Effects of large amounts of vitamin D3 injection on plasma vitamin D3 metabolites in lactating cows. PMID- 3033374 TI - Combination chemotherapy for unresectable hepatoblastoma in children. AB - Combination chemotherapy (adriamycin, vincristine, cyclophosphamide, etc.) was prescribed for two patients with initially unresectable hepatoblastoma. A significant reduction in tumor size occurred, and subsequent successful resection was feasible in one, although in the other the regressed tumor remained inoperable due to the link to the hepatic veins. The survival time for these patients was 9 months after operation and 41 months from diagnosis, respectively. Preoperative combination chemotherapy for initially unresectable hepatoblastoma facilitates surgical removal under more favorable conditions of a decreased tumor size. PMID- 3033376 TI - Induction of neutralizing antibodies by structural proteins VP1 and VP2 of swine vesicular disease virus. PMID- 3033377 TI - Establishment of a lymphoid cell line from tumor of bovine skin leukosis. PMID- 3033378 TI - Micro-method for neutralization test of transmissible gastroenteritis virus using porcine kidney cell line, CPK cells. PMID- 3033379 TI - Detection of rotaviruses in cat feces. PMID- 3033380 TI - The decline in the production of interleukin 2 in cats spontaneously infected with feline leukemia virus. PMID- 3033381 TI - Isolation of hemagglutinating encephalomyelitis virus from respiratory tract of pigs in Japan. PMID- 3033383 TI - Energy intake: its relationship to colon cancer risk. AB - A case-control study was conducted to assess the role of diet in the etiology of colon cancer. Diet was measured by means of a comprehensive quantifiable food frequency history instrument in 246 cases and 484 controls drawn from the general population of Utah. Each subject's diet was described by major nutrient groups and total energy based on the nutritional content of foods reported. Cases reported higher daily food intake 5 years preceding diagnosis than controls [men, rate ratio (RR) = 2.5; women, RR = 3.6], as measured by total energy content of the diet. Higher risk of colon cancer with increasing energy intake was independent of stage of disease at diagnosis and obesity, as measured by body mass. Fat, protein, and carbohydrate intake all had elevated RRs but could not be assessed as risk factors independent of energy intake because of their strong correlations with total calories. Due to the higher energy intake of the cases, odds ratios for the daily intake of dietary fiber and vitamins A and C were also greater than 1. However, adjusting for caloric intake removed this effect, and dietary fiber showed a weak protective effect. Total energy intake must be evaluated before attempting to assign a causal role to any food or nutrient that may be postulated to play a role in colon cancer. PMID- 3033384 TI - The role of HSV & HPV in cervical cancer. PMID- 3033382 TI - Integration and expression of provirus in human cells transformed by avian sarcoma virus. AB - Previously, human diploid fibroblasts from some donors infected in vitro by avian sarcoma virus (ASV) were transformed and found, by electron microscopy, to produce small numbers of virus particles that were infectious by bioassay; also, a line of human osteosarcoma cells infected with ASV developed additional characteristics of transformation and released a small number of infectious virus particles. In this study the complete proviral sequence was shown to be integrated in the genome of these cells. The env-related proteins gp85 and gp37 and the gag-related proteins pr76, pr60, and p19 can be detected in cytoplasmic extracts of ASV-infected human cells. Comparable amounts of pp60v-src were found in human and avian cells infected with ASV. The associated kinase activity in infected human cells was dramatically increased as compared to that of uninfected controls; the enzyme had the same cation and substrate requirements as those from ASV-transformed avian cells. Replicating particles from infected human cells were purified and were significantly modified compared to those from avian hosts as shown by a) higher specific gravity, b) the presence of RSV gag-related but not env-related antigens, and c) the fact that the virus-associated reverse transcriptase preferred the divalent cations Mn2+ and Fe2+ over Mg2+. PMID- 3033385 TI - [Certain current problems in health resort cardiology]. PMID- 3033386 TI - Aluminum action on mouse bone cell metabolism and response to PTH and 1,25(OH)2D3. AB - Aluminum (Al) accumulation in bone is associated with low bone formation and mineralization rates; resorption may also be reduced. The mechanism of these Al induced changes was investigated using cultured mouse osteoblast-like (OB) and osteoclast-like (OC) cells. The Al effect on bone resorption was measured by the in vitro release of 45Ca and beta-glucuronidase from mouse fetal limb-bones. Al had a biphasic effect. High concentrations (greater than 1.5 X 10(-6) M) of Al inhibited collagen and DNA synthesis, ornithine decarboxylase and alkaline phosphatase activity in OB, and depressed tartrate-resistant acid phosphatase activity in OC. Lower Al concentrations stimulated these cellular activities and 45Ca and beta-glucuronidase release from fetal bones. Al had no effect on basal cAMP levels in OB but inhibited the stimulating effect of bPTH on cAMP content. Al also altered the 1,25(OH)2D3 effects on the ornithine decarboxylase activity of OB cells. These data suggest that: (i) the low bone formation observed in vivo during Al intoxication may be due to the inhibition of collagen synthesis and to depressed cell proliferation; and (ii) Al may indirectly influence bone remodeling by interfering with the actions of bPTH and 1,25(OH)2D3 on bone cells. PMID- 3033387 TI - Angiotensin I converting enzyme and kinin-hydrolyzing enzymes along the rabbit nephron. AB - Angiotensin I converting enzyme (ACE) and kininase activities were measured in various segments of the rabbit nephron. ACE was determined with tritiated hippuryl-glycylglycine as substrate. Lysyl-bradykinin (LBK) hydrolysis (kininase activity) was measured by radioimmunoassay. ACE was only found in the glomerulus and in the two parts of proximal tubule: the convoluted proximal tubule and the pars recta (PR). It was distributed along a concentration gradient which increased from the glomerulus to PR. Kininase activity was found in both proximal and distal parts of the nephron. Besides intense LBK-hydrolyzing activity in the proximal tubule, a kininase activity was also found in the medullary collecting tubule (MCT). Kininase activity in the glomerulus and the proximal tubule was completely inhibited by chelating agents. Captopril inhibited this activity only in the PR and at high concentrations (above 10(-7) M). These results indicate that several types of enzymes other than ACE hydrolyze kinins in the glomerulus and in the proximal tubule. The contribution of ACE to kinin hydrolysis appears only minimal. The kininase activity found in MCT was different from ACE and other proximal tubule kininases because it was not inhibited by chelating agents. This kininase may play a physiological role in inactivating the kinins formed by kallikrein at or beyond the connecting tubule. PMID- 3033388 TI - Reversing glomerular hypertension stabilizes established glomerular injury. AB - Munich-Wistar rats were studied 18 weeks following 5/6 renal ablation. In untreated group 1 rats maintained on standard chow containing 24% protein, sustained systemic and glomerular hypertension were associated with increasing proteinuria and widespread glomerular injury. In group 2, early treatment with the converting enzyme inhibitor enalapril prevented systemic and glomerular hypertension, and largely limited proteinuria and glomerular injury. Groups 3 and 4 received no therapy during the first eight weeks, during which they developed systemic hypertension and levels of proteinuria previously shown to be associated with significant glomerular sclerosis at this time point. Enalapril therapy begun at eight weeks in group 3 rats reversed systemic and glomerular hypertension, prevented a further rise in proteinuria, and limited glomerular lesions at 18 weeks relative to group 1. Reduction of dietary protein content to 12% at eight weeks in group 4 rats controlled glomerular but not systemic hypertension to near normal levels, stabilized proteinuria values, and also limited glomerular lesions at 18 weeks compared to group 1. These studies support the view that glomerular hypertension is an essential hemodynamic derangement responsible for progressive glomerular injury. Furthermore, reduction of capillary pressure can arrest the progression of remnant glomerular injury even when therapy is delayed until glomerular injury is established. PMID- 3033389 TI - Moderate sodium restriction in hypertensive subjects: renal effects of ACE inhibition. AB - It has been suggested that AII-mediated renal mechanisms limit the efficacy of moderate sodium restriction in the lowering of blood pressure (BP) in hypertension. We therefore studied renal hemodynamics and sodium handling in nine essential hypertensives in balance on 200 and on a 50 mmol sodium diet, before and during ACE-inhibition (enalapril 10 mg bid for 8 days) in a cross-over fashion. BP was similar on 50 and 200 mmol Na before enalapril, the fall in BP during enalapril was significantly more pronounced on 50 mmol Na. On 50 mmol Na, GFR and filtered Na were significantly lower, and tubular reabsorption was significantly higher than on 200 mmol Na. GFR increased during enalapril in 50 but not on 200 mmol Na. Consequently, the differences in GFR and filtered load elicited by sodium restriction were no longer present during ACE-inhibition. In contrast, the differences in tubular reabsorption between 50 and 200 mmol Na persisted during enalapril. In conclusion, moderate sodium restriction, not affecting BP, can elicit a renal hemodynamic response. As this response is blunted by ACE-inhibition it is probably mediated by AII. This blunting may contribute to the increased sodium sensitivity of BP during ACE-inhibition. The adaptation of tubular sodium reabsorption is not affected by ACE-inhibition. PMID- 3033390 TI - [Dispensarization of inguinal hernia patients]. PMID- 3033391 TI - [Hypophyseal and steroid hormones in women with breast cancer]. PMID- 3033392 TI - [Morphologic studies of the pathogenesis of ligneous conjunctivitis]. AB - The cases of ligneous conjunctivitis published since 1964 are presented in a review of the literature. A total of 84 cases have been described. The predominance of this disease in women is lower than was previously thought. Ligneous conjunctivitis tends to occur more frequently in children. However, it is not a "disease of little girls". Two cases are described in the present paper, including pathomorphological studies. In both cases, neutrophilic granulocytes were studied by electron microscopy. The results are compared with a control case and with corresponding reports in the literature. The number of granules in the neutrophilic granulocytes was significantly reduced in both cases. In one of the cases studied the granules were increased in size. These findings support the hypothesis that ligneous conjunctivitis could be the result of a lowered resistance associated with a disturbance of wound healing. PMID- 3033395 TI - [The role of dietary fiber in nutrition]. PMID- 3033393 TI - Alteration of oral carbohydrate tolerance during administration of a fiber-free formula diet. AB - Fiber-free formula diets are widely used for medical and experimental purposes. In healthy individuals no major changes of fasting blood glucose have been reported, whereas oral glucose tolerance tests indicate a deterioration of carbohydrate tolerance following ingestion of liquid diets containing certain sugars. We examined carbohydrate tolerance in normal subjects during prolonged administration of a fiber-free semisolid diet containing polysaccharides instead of sugars. In the first experiment, diet A (55% starch, 40% highly polyunsaturated fat, P/S 5.4, 15% protein) served as the sole source of energy for 4 weeks. During this time a progressive deterioration of carbohydrate tolerance was observed, indicated by a gradual increase of the postprandial glycemic response (area under the curve) from -316 to +3874 mg/dl X min without significant alteration of postprandial serum insulin concentrations. For the second study, nutrient relations of the diet were modified (diet B: 45% starch, 40% saturated fat, P/S 0.3, 15% protein). The changes of carbohydrate tolerance during 1 week of feeding diet B were similar to those seen in the first experiment. However, follow-up observations during a subsequent pharmacological trial showed that the changes of postprandial blood glucose curves were reversed within 2 weeks. This was associated with an enhanced serum insulin response. The reason for the different durations of the metabolic effects of diets A and B are not obvious from our data. The time course may have been influenced by the different nutrient intake. Fasting blood glucose concentrations were not sensitive enough to detect the deterioration of carbohydrate tolerance. PMID- 3033394 TI - Naloxone in treatment of circulatory shock resistant to conventional therapy. AB - The effect of naloxone (4.4-5.9 mg i.v.) was evaluated in 10 patients with circulatory shock (sepsis, n = 7; intoxication, n = 1; cardiogenic shock, n = 2) not responding to full conventional therapy. In addition, we measured plasma ACTH and immunoreactive beta-endorphin before and 60 min after administration of naloxone and compared the results with hormone concentrations in 10 intensive care patients without shock. Only in two patient with septic shock a transient increase (duration 15 min and 60 min, respectively) of systolic blood pressure was observed, while naloxone was ineffective in the remaining eight patients. No adverse effects of naloxone were found. Plasma ACTH and immunoreactive beta endorphin concentrations in patients with shock were not different from those in controls (ACTH, 79 +/- 28 vs 120 +/- 60 pg/ml; immunoreactive beta-endorphin, 952 +/- 262 vs 1,070 +/- 378 pg/ml). Our findings suggest that naloxone in a single dose of 4.4-5.9 mg i.v. does not improve the management of circulatory shock unresponsive to conventional treatment. beta-endorphin seems to play no major role in the hypotension of shock. PMID- 3033397 TI - Detection of past and recurrent marijuana use by a modified GC/MS procedure. AB - A published gas chromatography/mass spectrometry method for detecting 11-nor-9 carboxy-delta-9-tetrahydrocannabinol (THC-COOH) in urine was modified. In the new procedure, five GC/MS ion peaks of a pentafluoropropylpentafluoropropionyl derivative of THC-COOH are monitored in the multiple-ion mode for improved reliability; two ion peaks of the trideuterated internal standard are used for greater quantitative precision; and methanolic KOH is employed instead of NaOH in the hydrolysis and extraction steps to produce a cleaner extract. Finally, the new procedure is designed to keep THC-COOH either in basic solution or in an organic solvent at all times to prevent adsorption onto glass or plastic, so that disposable, non-silanized glassware may be used. Patient urines were analyzed by both the new procedure and the EMIT method. For 32 specimens, the average and range of EMIT/GC/MS concentration ratios were 2.8 and 0.9-7.2, respectively. Concentrations of THC-COOH measured by the GC/MS procedure may be more indicative of recent marijuana use than the EMIT semi-quantitative concentration values. PMID- 3033396 TI - Regional distribution of the phenylalanine-sensitive ATP-sulphurylase in brain. AB - The regional distribution of the phenylalanine-sensitive ATP-sulphurylase in fetal calf brain coincides with demyelinated lesions observed in the central nervous systems of untreated PKU patients. This would be expected if this species of ATP-sulphurylase played a role in the pathogenesis of brain dysfunction in the untreated or poorly controlled phenylketonuria patient. PMID- 3033398 TI - Additional results in the validation of the Bond Elut-THC extraction column--TLC as a confirmation method for EMIT- and RIA-positive cannabinoid urines. PMID- 3033400 TI - Epidemiology and clinical characteristics of testicular tumors in Saudi Arabia: King Faisal Specialist Hospital and Research Centre experience. AB - The data on 62 patients with germinal testicular tumors seen at King Faisal Specialist Hospital and Research Centre between January 1977 and June 1983 were analyzed to determine the epidemiologic and clinical characteristics of the disease in Saudi Arabia. Fifty-seven patients were Saudis. The geographic distribution of Saudi patients seemed to coincide with population concentrations. Testicular seminomas (TS) and non-seminomatous testicular tumors (NSTT) comprised 50% each. The mean age was 41 and 27.8 years for TS and NSTT, respectively. Fifteen patients had cryptorchidism of the involved testicle. Three patients with NSTT had a history of trauma to the involved testicle. The most common presentations were painless testicular swelling (51.6%), painful swelling (16%), and abdominal or inguinal swelling (21%). The delay between the onset of symptoms and referral (mean 15 months) was considerable. Eighty percent of patients with NSTT and 45% of those with TS had advanced disease at referral. PMID- 3033399 TI - A specific histamine-stimulated phosphoprotein in isolated parietal cells. AB - Histamine-stimulated phosphorylation was studied in isolated rabbit parietal cells. Secretion of acid, as assessed by aminopyrine uptake, was linear at 15 min of stimulation with histamine. By utilizing two dimensional gels, a specific 30,000-Da protein (pp30) was identified whose phosphorylation was prominently stimulated by histamine after 15 min of incubation. The pp30 protein displayed an isoelectric point of 6.0. Furthermore, cAMP-dependent pp30 phosphorylation could also be demonstrated in vitro in a preparation of parietal cell cytosol. The results suggest that pp30 may represent an important histamine-stimulated cAMP dependent phosphoprotein involved in the initiation or maintenance of parietal cell secretion. PMID- 3033401 TI - Changing incidence of adenocarcinoma of the lung: a brief review. AB - The autopsy of lung cancer at Hines Veterans Administration Hospital was reviewed for a 27-year period, between 1957 and 1983. There has been a changing histologic pattern with a recent increase in adenocarcinoma and a relative decrease of the other cell types. This observation, based on a virtually all-male patient population, is partly attributed to better and improved diagnostic procedures. PMID- 3033402 TI - Relationship of nuclear appearance to stromal invasion in human breast cancer. AB - Two biopsies of intraductal and invasive lobular carcinoma of the breast and 11 biopsies of intraductal and invasive carcinoma of the breast were examined by automated micromensurative techniques for mean nuclear area of carcinoma cells in intraductal and in invasive compartments. The nuclei of invasive carcinoma cells tended to be smaller when they invaded stroma as single cells or as "thin strands" than when they invaded as part of large sheets of cells. Cells in direct apposition with the stroma usually had smaller nuclei than cells which were found centrally in large confluent sheets of cells. Intraduct carcinoma cells approached, in size, invasive cells which formed confluent sheets, rather than cancer cells which invaded as single cells. The nuclei of cells which invaded in small groups were usually darker than those of cells in confluent sheets, which were usually more vesicular. PMID- 3033403 TI - Establishment of bovine leukemia virus-producing and -nonproducing B-lymphoid cell lines and their proviral genomes. AB - We have established four cell lines in vitro from peripheral leukemic cells of four independent enzootic bovine leukosis (EBL) cattle. All cell lines exhibited differentiated B-cell characters. Two of them produced infectious bovine leukemia virus (BLV), but the others did not. Nonproducer cell lines contained single copies of defective BLV proviral genomes with the same integration profiles as the uncultured cells. On the other hand, numerous proviral copies were detected in producer cell lines. One of the producer cell lines, BL407, whose original uncultured cell contained complete and defective proviral genomes retained the original two copies and had increased only complete genomes in different integration sites after long term culture. These findings suggest that the monoclonal leukemic cells from EBL cases are preferentially established in vitro irrespective of their proviral structures, and the producer B-lymphoid cells amplify their proviral copies by reinfection with viruses re-expressed from the cells during in vitro cultivation. PMID- 3033404 TI - Intracellular recording of identified dorsal spinocerebellar tract neurones from guinea pig spinal cord in vitro. AB - Recent studies on the monosynaptic connection between primary afferent fibres and dorsal spinocerebellar tract (DSCT) neurones in the spinal cord of anaesthetized cats have been undertaken to investigate the mechanisms of excitatory synaptic transmission in the mammalian central nervous system. The need to extend these observations to a study of the effects of changes in the extracellular environment of DSCT neurones prompted us to develop the in vitro preparation described in this paper. We have developed an isolated spinal cord preparation from young guinea pigs, in which DSCT neurones can be identified and recorded from intracellularly. The spinal cord can be maintained in vitro for at least 7 hours. This preparation is sufficiently stable to allow intracellular penetration of DSCT cells in excess of 2 h, with resting membrane potentials of -65 mV obtainable. Reconstruction of neurones stained with horseradish peroxidase confirmed their location in Clarke's column. The dendritic morphology of the reconstructed DSCT neurones was found to be similar to DSCT neurones described in the cat spinal cord. Since the motor system of the guinea pig is quite advanced at birth, this preparation should provide a valid extension to in vivo results on neuronal membrane properties and synaptic potentials in DSCT neurones. PMID- 3033406 TI - [Paraneoplastic retinal degeneration]. PMID- 3033405 TI - [Undifferentiated small cell carcinoma of pancreatic origin. Presentation of a case and review of the literature]. PMID- 3033407 TI - [Cytomegalovirus rhabdomyolysis]. PMID- 3033408 TI - [Hepatic encephalopathy in a 57-year-old male with Caroli's disease]. PMID- 3033409 TI - Synthesis of vitamin D3 and related compounds. PMID- 3033410 TI - The release of phosphate from contracting atria as a parameter of (Na+ + K+) ATPase activity. Effect of ouabain. AB - An experimental situation that permits simultaneous evaluation of the mechanical activity and inorganic phosphate (Pi) release by beating atria was developed. It concerned cell membrane integrity and compartmentalization of the cells. The Pi released by isolated rat and guinea pig atria was measured in a physiological salt solution. The preparations were incubated under preload for the simultaneous recording of dF/dt and Pi. The substrate of this reaction was the endogenous ATP. Pi release by atria increased with time. It was stimulated at high K+ concentrations (30 mmol/l) and inhibited at very low ones (1.2 mmol/l). The lack of Mg2+ in the incubation media did not affect it. Ouabain induced different effects upon Pi release; at low concentrations it was increased while at higher ones it was inhibited. The relationship between ouabain concentrations, mechanical responses and liberation of Pi by isolated atria from rat and guinea pig showed that at low concentrations ouabain simultaneously increased dF/dt and Pi release. On the contrary, at high concentrations of ouabain dF/dt decreased, contracture occurred and the Pi release decreased. Isoproterenol, norepinephrine and calcium, at concentrations that increased dF/dt, did not modify Pi release. Different concentrations of potassium and ouabain also modulated the 32P Pi efflux in a fashion similar to that with the Pi released by beating atria. We propose that Pi release by living cells appears to be correlated with manipulations which either stimulate or inhibit (Na+ + K+)-ATPase activity. PMID- 3033411 TI - Evidence of beta-adrenergic involvement in forced swimming-induced behavioural despair of mice. AB - Modulation of forced swimming-induced immobility by beta-adrenoceptor activation was investigated in mice. Isoprenaline prolonged the immobility duration of mice in a dose-related manner when the animals were forced to swim for 6 min periods. On the other hand, salbutamol (a beta 2-agonist) reduced the immobility duration of mice. Pretreatment with propranolol (1,2,4,8 and 16 mg/kg, ip), atenolol (10 mg/kg, ip) and metoprolol (10 mg/kg, ip) antagonized the immobility-enhancing effect of isoprenaline. Chronic administration (10 mg/kg/day) for 8 days of propranolol and imipramine protected the animals from isoprenaline-evoked prolongation of immobility. These findings suggest that central beta 1- and beta 2-subtypes of adrenoceptors may be acting in opposite directions to modify the immobility duration of mice. Activation of central beta 1-adrenoceptors may lead to enhanced behavioural despair, whereas a reverse effect may be observed on beta 2-adrenoceptor activation. PMID- 3033412 TI - Strategies in the development of new drugs and drug combinations against leprosy, demonstrated on the example of folate and gyrase inhibitors. PMID- 3033413 TI - Selective interaction of tricyclic antidepressants with a subclass of rat brain cholinergic muscarinic receptors. AB - Experiments were undertaken to determine whether the anticholinergic actions of tricyclic antidepressants are mediated by a selective interaction with a subclass of muscarinic receptors. To this end, the potencies of these antidepressants to inhibit [3H]-QNB binding to rat brain cerebral cortical membranes was compared to their potencies as antagonists of carbachol-stimulated inositol phosphate accumulation in cerebral cortical slices and carbachol-induced inhibition of GTP stimulated adenylate cyclase in striatal membranes. Whereas amitriptyline was more potent than pirenzepine, a selective muscarinic M1 receptor antagonist, in competing for [3H]-QNB binding sites and as an antagonist of carbachol-induced inhibition of adenylate cyclase, pirenzepine was substantially more active (ten fold) than amitriptyline in blocking carbachol-stimulated phosphatidyl inositol turnover. Atropine was more potent than all other agents in these assays, failing to display any significant degree of selectivity. The results suggest that the tricyclic antidepressants, in particular amitriptyline, appear to be selective antagonists for muscarinic receptors associated with adenylate cyclase in striatal membranes. Given the current classification of cholinergic receptors, these findings indicate that the tricyclic antidepressants may be useful for defining the properties of M2 receptors in brain. PMID- 3033414 TI - Coupling of adrenal medullary opioid receptors to islet-activating protein sensitive GTP-binding proteins. AB - Possible coupling of bovine adrenal medullary opioid receptors to islet activating protein (IAP, pertussis toxin)-sensitive GTP-binding proteins was investigated by studying effects of guanyl-5'-yl imidodiphosphate (Gpp(NH)p) and IAP treatment of membranes on opioid binding. Gpp(NH)p inhibited [3H]D-Ala2-D Leu5-enkephalin ([3H]DADLE) binding by increasing the dissociation constant of [3H]DADLE and membranes, and enhanced slightly [3H]diprenorphine binding. IAP treatment of membranes reduced [3H]DADLE binding and abolished almost completely the Gpp(NH)p inhibition of [3H]DADLE binding. Treatment of membranes with IAP and [32P]NAD resulted in radio-labeling of membrane proteins of approximately 39,000 dalton. DADLE inhibited adenylate cyclase activity in rat brain caudate nucleus. However, DADLE, beta-endorphin, levorphanol and dynorphin A(1-13) did not show any significant inhibitory action on bovine adrenal medullary adenylate cyclase activity. These results suggest that bovine adrenal medullary opioid (DADLE) receptors are linked to IAP-sensitive GTP-binding proteins which are not directly coupled to adenylate cyclase. PMID- 3033415 TI - Facilitation of nerve stimulation evoked noradrenaline overflow by isoprenaline but not by circulating adrenaline in the dog in vivo. AB - The influence of isoprenaline and adrenaline on the overflow of endogenous noradrenaline evoked by sympathetic nerve stimulation was studied in canine blood perfused gracilis muscle in situ. Neuronal uptake was inhibited by desipramine. Local i.a. infusions of isoprenaline enhanced stimulation evoked noradrenaline overflow by 32 +/- 10% (P less than 0.05), indicating the existence of prejunctional facilitatory beta-adrenoceptors. This effect of isoprenaline was not antagonized by beta 1-adrenoceptor blockade and does not seem to be related to the vasodilatation caused by isoprenaline. In a second series of experiments circulating adrenaline levels were raised by i.v. infusions from basal levels of 0.4 +/- 0.2 nM to 1.7 +/- 0.2 and 6.3 +/- 0.6 nM, respectively, in arterial plasma. Adrenaline elicited vasodilatation in the gracilis muscle (19 +/- 3 and 28 +/- 5% increases in vascular conductance, respectively), indicating activation of postjunctional beta 2-adrenoceptors, without influencing nerve stimulation evoked noradrenaline overflow. Thus, our results support the existence of a prejunctional beta 2-adrenoceptor mediated mechanism facilitating noradrenaline release in vivo, but provide no evidence to support the idea that physiologically relevant increases in circulating adrenaline levels enhance noradrenergic neurotransmission in skeletal muscle. PMID- 3033416 TI - Regulation of cyclic-AMP synthesis in amphibian melanotrope cells through catecholamine and GABA receptors. AB - Catecholamines and GABA are neurotransmitters involved in the regulation of release of pro-opiomelanocortin (POMC) derived peptides from the neurointermediate lobe of Xenopus laevis. The present study concerns the relation of these neurotransmitters to the adenylate cyclase system of the melanotrope cell. During in vitro incubation of isolated melanotrope cells it was found that dopamine, adrenaline and LY 171555 induced inhibition of forskolin-stimulated cAMP production and concomitantly inhibited MSH release. Activation of the GABAb receptors by baclofen also induced inhibition of cAMP production and alpha MSH secretion. Activation of the GABAa receptors evoked stimulation of cAMP production, while alpha MSH release was slightly inhibited, indicating that the GABAa mechanism may prove to be complex. A dual regulation through two subtypes of this receptor might be involved, one stimulating release through the adenylate cyclase system, while the other would inhibit secretion. PMID- 3033417 TI - The benzodiazepine agonist clonazepam potentiates the effects of gamma aminobutyric acid on alpha-MSH release from neurointermediate lobes in vitro. AB - The action of the central-type benzodiazepine-receptor agonist clonazepam on alpha-MSH release has been studied in vitro using perifused frog neurointermediate lobes. High concentrations of clonazepam (3.16 X 10(-5) and 10( 4) M) caused an inhibition of alpha-MSH release and this effect was reversed by the central-type benzodiazepine-receptor antagonist Ro 15-1788. High doses of GABA (10(-5) and 10(-4) M) induced a biphasic effect on pars intermedia cells: a brief stimulation followed by a sustained inhibition of alpha-MSH secretion. Administration of clonazepam (10(-5) M) in the presence of various concentrations of GABA (10(-6) to 10(-4) M) led to a potentiation of both stimulatory and inhibitory phases of alpha-MSH secretion induced by GABA. Ro 15-1788 completely abolished the potentiating effect of clonazepam. Thus our results indicate that endogenous benzodiazepine receptors may modulate the effects of GABA on alpha-MSH secretion. PMID- 3033418 TI - C57BL/6J-bgJ (beige) mice: differential sensitivity in the tail flick test to centrally administered mu- and delta-opioid receptor agonists. AB - The antinociceptive effects of two mu-opioid receptor agonists, morphine and [D Ala2, MePhe4, Gly-ol5]enkephalin (DAGO), and a selective delta-receptor agonist, [D-Pen2, L-Pen5]enkephalin (DPLPE), were determined in C57BL/6J-bgJ (beige) and control mice (CRS-CDl and C57BL/6By) using a standard tail-flick assay. The antinociceptive response of C57BL/6J-bgJ mice to intracerebro-ventricularly administered morphine and DAGO was significantly reduced compared to controls, but there was no difference in the antinociceptive response to DPLPE. These results suggest that there is a genetic deficit of mu-opioid receptor number or a genetically-induced alteration in receptor function in regions of C57BL/6J-bgJ brains involved in antinociception, that delta-opioid receptors can mediate antinociception in mice, and that the C57BL/6J-bgJ strain may offer a practical new animal model for studying the function of opioid receptor subtypes. PMID- 3033419 TI - The formation of lysophosphatidylinositol phosphate in human platelet microsomes. AB - The formation of lysophosphatidylinositol phosphate (lysoPIP) from lysophosphatidylinositol (lysoPI) via kinase activity was studied in microsomal preparations from human platelets. For this purpose, [3H]lysoPI or [3H]phosphatidylinositol ([3H]PI) was prepared and incubated in the presence or absence of ATP, MgCl2 and Triton X-100, and the appearances of radioactivity in [3H]lysoPIP and [3H]phosphatidylinositol phosphate ([3H]PIP), respectively, were monitored using thin layer chromatography. Both lysoPI and PI phosphorylations were completely dependent upon the presence of ATP and MgCl2 in the incubation medium; Triton X-100 addition stimulated both reactions, with the stimulation of PI conversion being considerably greater than that for lysoPI can be converted to lysoPIP by phosphorylation in human platelet microsomes. The potential significance of this enzymatic reaction in stimulated cells is discussed in relation to the generation of inositol-1,4,5-trisphosphate, an important intracellular second messenger. PMID- 3033421 TI - Ground squirrel hepatitis virus (GSHV) infection and hepatocellular carcinoma in the Canadian Richardson ground squirrel (Spermophilus richardsonii). AB - Sera and livers from 40 Richardson ground squirrels were examined for evidence of ground squirrel hepatitis virus (GSHV) infection and hepatocellular carcinoma. Twenty-five sera were obtained from fully grown adult ground squirrels and 15 from young ground squirrels estimated to be between 6-8 weeks of age. All animals had been caught in the wild and had spent less than 1 month in captivity. Sixteen sera (40%) had at least one serologic marker of GSHV (2 with GSHV surface antigen, 3 GSHV core antigen, 5 GSHV antibody to core antigen and 11 GSHV antibody to surface antigen). Two cases of hepatocellular carcinoma were detected, both in adult ground squirrels. Tumour tissue and adjacent normal liver tissue were negative for GSHV surface and core antigen by direct immunofluorescence in both livers and negative for GSHV-DNA by molecular hybridization in the one tumour examined. These results indicate that: A) GSHV is geographically more widespread than previously considered; B) viral transmission occurs early in life, probably via the vertical route; and C) hepatocellular carcinoma is a relatively common finding in these animals while still in their wild state. Any causal relationship between hepatocellular carcinoma and GSHV infection in these animals, however, has yet to be demonstrated. PMID- 3033420 TI - Two new icosapentaenoic acids from the temperate red seaweed Ptilota filicina J. Agardh. AB - Two new fatty acid metabolites, 5(Z),7(E),9(E),14(Z),17(Z)-icosapentaenoic acid and 5(E),7(E),9(E),14(Z),17(Z)-icosapentaenoic acid, have been isolated from the temperate red marine alga, Ptilota filicina (Ceramiales, Rhodophyta). The structures of these new compounds, isolated as their methyl ester derivatives, have been deduced from detailed 1H nuclear magnetic resonance (NMR), 13C NMR and 2D-NMR analyses as well as comparisons to known compounds. PMID- 3033422 TI - Relationship of histologic grade of hepatocellular carcinoma (HCC) to tumor size, and demonstration of tumor cells of multiple different grades in single small HCC. AB - The histologic features of 65 surgically resected cases of hepatocellular carcinoma (HCC) were studied, and the cellular differentiation was graded from I to IV according to Edmondson-Steiner's classification. HCC cells of 2 or more grades were seen in 31 (47.7%) of the 65 cases. There was no significant difference in the proportion of HCC composed of tumor cells of more than one histologic grade between HCCs with association with liver cirrhosis and without cirrhosis. Among these 31 cases, extremely well-differentiated HCC, corresponding to Edmondson-Steiner's grade I carcinoma, was found in 9 of 11 tumors smaller than 3 cm in diameter, but it was not seen in 20 tumors larger than 3 cm in diameter. In the 34 HCCs with a uniform histologic pattern, all of two minute tumors smaller than 1 cm in diameter consisted of extremely well-differentiated HCC, but there were no cases consisting of extremely well-differentiated HCC in tumors larger than 2 cm in diameter. Taken together, these findings suggest that HCCs originate as relatively well-differentiated tumors, which may be difficult to distinguish from adenomatous regenerative nodules, and become progressively less differentiated at a later stage of their development. PMID- 3033423 TI - Spontaneous and antibody-dependent cellular immune reactions to ethanol-altered hepatoma cells. AB - Spontaneous cell-mediated cytotoxicity (SCMC), antibody-dependent cellular cytotoxicity (ADCC) and proliferative lymphocyte stimulation in alcoholic liver disease (ALD) were investigated. Peripheral blood lymphocytes (PBL) from eight patients with advanced ALD and nine normal controls were tested against hepatoma cells (PLC/PRF/5) as targets. Target cells were grown in either normal culture medium or medium supplemented with 1 and 5% ethanol, respectively, for 24 to 48 h. Ethanol-exposed hepatoma cells exhibited profound and characteristic morphological alterations. Ethanol preincubation, however, proved to be without effect on immune reactions. Provided that hepatoma cells are an appropriate model, we assume that the proposed immune reactions in ALD are based on metabolic interactions operative only in vivo but do not parallel morphological alterations of liver cells directly induced by ethanol. PMID- 3033425 TI - Echo acquisition during frequency-selective pulse trains for proton spectroscopy of metabolites in vivo. AB - Suppression of water and fat in proton spectra of rat skeletal muscle was achieved by acquisition of the spin echoes formed in the steady state after a series of frequency-selective composite pulses of constant amplitude, phase, and interpulse interval. The method enables observation of lactate in the presence of fat without resorting to difference spectroscopy, and on spectrometers which permit data acquisition immediately prior to a pulse, enables much faster data accumulation compared to standard echo sequences. PMID- 3033424 TI - In vivo 31P NMR spectroscopy of agonist-stimulated phosphatidylinositol metabolism in cat brain. AB - As a result of the agonist stimulation of muscarinic receptors, myo-inositol-1 phosphate accumulates in the presence of millimolar lithium concentrations. This accumulation of myo-inositol-1-phosphate can be detected by in vivo 31P NMR spectroscopy. Since myo-inositol-1-phosphate is a breakdown product of phosphatidylinositol, this may provide a means of noninvasively monitoring phosphatidylinositol metabolism in vivo. PMID- 3033426 TI - [Serum concentration of alpha 1-antitrypsin in various hepatic diseases]. PMID- 3033427 TI - [Latent infection and recurrent disease caused by herpes simplex type 1: prophylactic and therapeutic perspectives]. PMID- 3033428 TI - [The epidemic of AIDS (acquired immunodeficiency syndrome)]. PMID- 3033429 TI - [Immunofluorescence in the serological study of HTLV-III/LAV]. PMID- 3033430 TI - [Undifferentiated small cell carcinoma (oat cell). Review of 55 cases, including 4 extrapulmonary]. PMID- 3033431 TI - Assays for superoxide dismutase. PMID- 3033432 TI - Phospholamban involvement in the maintenance of basal calcium transport in cardiac sarcoplasmic reticulum. AB - Phosphorylation of cardiac sarcoplasmic reticulum membrane vesicles by exogenous c-AMP and c-AMP-dependent protein kinase stimulates calcium uptake and Ca2+ dependent ATP hydrolysis by 40-50% and results in the incorporation of 32P into a 22-KDa protein, phospholamban. Treatment of the membrane with DOC (0.0002% or 5 X 10(-6) M) solubilizes phospholamban from the membrane and induces a 90% inhibition of basal calcium uptake. This inhibition cannot be attributed to an alteration in vesicle integrity or membrane permeability. The (Ca2+ + Mg2+) ATPase remains associated with the membrane fraction and exhibits optimal levels of Ca2+-stimulated ATP hydrolysis. Phosphorylation prior to DOC treatment allows retention of the phospholamban in the membrane, concomitant with maintenance of the calcium transport activity. The results presented suggest that phospholamban is involved in the maintenance of basal calcium transport function in cardiac sarcoplasmic reticulum and that its phosphorylation stimulates Ca2+ transport. PMID- 3033433 TI - Peptide transport in Salmonella typhimurium: molecular cloning and characterization of the oligopeptide permease genes. AB - The oligopeptide permease is encoded by at least four genes which are transcribed as a single operon. We cloned and characterized this operon from Salmonella typhimurium, as well as the flanking genes, tonB, ana and a new gene, cwd, which affects cell wall synthesis. We correlated the physical map of opp DNA with a detailed genetic map of the opp operon and the individual opp genes were accurately located with respect to various restriction sites by Southern blotting. The region of the chromosome near opp was found to be highly unstable with deletions arising at a highly frequency. The operon also contains hot-spots for IS1 and IS5 insertions. PMID- 3033434 TI - Linear DNA must have free ends to transform rat cells efficiently. AB - We have observed that failure to remove certain restriction enzymes after digestion reduced the transforming ability of DNA from 10- to 50-fold. The DNA found integrated in the transformed cells isolated under these conditions had lost little or no sequences. We interpret these results as indicating that certain restriction enzymes remain bound to the DNA ends after digestion, thus generating a substrate unfavorable both for integration and exonucleolytic degradation. As expected from this interpretation, removal of the restriction enzymes before transfection restored the full transforming ability of linear DNA, but also resulted in the integrated sequences being significantly shorter than the transfected DNA. These findings strongly argue for the hypothesis that integration of linear DNA by illegitimate recombination requires free ends and further suggest that exonucleolytic degradation of such ends may generate a preferred substrate for integration. Finally, a comparison of the sequences found integrated after transfection with circular or linear molecules, led us to conclude that circular molecules need not be linearized to become integrated. PMID- 3033435 TI - Isolation of the yeast phosphoglyceromutase gene and construction of deletion mutants. AB - The PGM1 gene (also called GPM; Fraenkel 1982) coding for phosphoglyceromutase was isolated by functional complementation. When present on a multicopy vector and introduced into yeast cells it led to an about eightfold increase in specific enzymatic activity. This apparent overproduction was confirmed by SDS polyacrylamide gel electrophoresis of crude extracts and at the transcriptional level by Northern analysis. By subcloning of the yeast DNA insertions of the plasmids originally isolated the PGM1 coding region was located within a 1.3 kb SalI-HindIII fragment. Integration at the chromosomal locus confirmed that the PGM1 gene had indeed been isolated. Southern analysis of genomic digests showed the same restriction patterns as the cloned sequences. However, a BamHI restriction polymorphism was observed. Furthermore, a repetitive element was found in the PGM1 flanking region. Finally, the chromosomal copy of the gene was deleted by replacement with a URA3 marker. The deletion mutants showed that the gene is not essential for yeast growing in the presence of a combination of glycerol and ethanol. However, growth was inhibited by glucose and neither glycerol nor ethanol alone were sufficient to support growth. PMID- 3033436 TI - Cloning in Escherichia coli of genes involved in the synthesis of proline and leucine in Desulfovibrio desulfuricans Norway. AB - A library of Desulfovibrio desulfuricans Norway genomic DNA was constructed in Escherichia coli with pBR322 as vector and plasmids able to complement the proA and leuB mutations of the host were screened. It was observed that all the plasmids studied were highly unstable, the insert DNA being rapidly lost under non-selective growth conditions. A 2.75 kb DNA fragment of D. desulfuricans Norway was found to complement E. coli ProA, ProB and ProC deficiencies. From the results of restriction analysis and Southern hybridizations, it is proposed that the genes involved in proline and leucine biosynthesis are clustered on the chromosome of D. desulfuricans Norway. PMID- 3033437 TI - Mobilization of the non-conjugative plasmid RSF1010: a genetic analysis of its origin of transfer. AB - The oriT site of the broad host-range multicopy IncQ plasmid RSF1010 was cloned onto the 2.2 kb pBR322-derived vector pED825. By successive subcloning and construction of deletions, the oriT region was localised on an 80-88 bp segment of DNA. This segment was contained within the HaeII fragment of RSF1010 that is known to include the relaxation nick site. The oriT region was sequenced and inverted repeats and sequences homologous to the oriT regions of ColE1 and RK2 were identified. A striking 10 bp inverted repeat at one end of the 88 bp oriT segment may be important for recognition of oriT, and its possible role in transfer is discussed. As for other plasmids, the oriT region served as the site for recA-independent, transfer-dependent, site-specific recombination. This provides genetic evidence that strand breakage and re-joining occur at oriT during transfer. Mobilization was independent of transcription by RNA polymerase in the donor cell, as shown by the lack of effect of rifampicin. Inversion of the oriT site with respect to the plasmid oriV site showed that there was no functional dependence of oriT on oriV for synthesis of primers possibly involved in recipient conjugal DNA synthesis. Alternative mechanisms are discussed. PMID- 3033438 TI - Mobilization of the non-conjugative plasmid RSF1010: a genetic and DNA sequence analysis of the mobilization region. AB - The entire region required for mobilization of the non-conjugative plasmid RSF1010 has been cloned into a mobilization-deficient pBR322 derivative. The segment of DNA cloned was approximately 1.8 kb and included the origin of conjugal DNA transfer (oriT). The DNA sequence of the mobilization region has been determined, and revealed the presence of several overlapping reading frames. The isolation and mapping of both Tn1725 and BamH1-linker insertions and comparison with the DNA sequence data has allowed the identification of three genes required for mobilization. Two of these genes are overlapping and encode proteins of 16 kDa and greater than 65 kDa (although the truncated protein is functional, the gene extends outside the region cloned). The third gene is transcribed in the opposite direction. Promoters capable of transcribing these genes were located by S1 mapping in the inter-cistronic region between these divergently transcribed genes. The oriT site is located in this region, and the transcriptional patterns observed for mob+ and mob- plasmids implied that the promoters may be regulated by two of the mobilization proteins binding to the oriT site. PMID- 3033439 TI - The glucose kinase gene of Streptomyces coelicolor and its use in selecting spontaneous deletions for desired regions of the genome. AB - A number of deletions in the glucose kinase (glk) region of the Streptomyces coelicolor chromosome were found among spontaneous glk mutants. The deletions were identified by probing Southern blots of chromosomal DNA from glk mutants with cloned glk DNA. The deletions ranged in size from 0.3 kb to greater than 2.9 kb. When cloned glk DNA was introduced on a phi C31 phage vector into a glk mutant that contained a deletion of the entire homologous chromosomal glk region, glucose kinase activity was detected in extracts of these cells. The entire coding information for at least a subunit of glucose kinase is therefore present on the cloned glk DNA. The 0.3 kb glk chromosomal deletion was used to demonstrate that transfer of chromosomal glk mutations on to the phi C31::glk phage could occur by recombination in vivo. Since glk mutations frequently arise from deletion events, a method was devised for inserting the cloned glk DNA at sites in the chromosome for which cloned DNA is available, and thus facilitating the isolation of deletions in those DNA regions. phi C31::glk vectors containing a deletion of the phage att site cannot lysogenize S. coelicolor recipients containing a deletion of the glk chromosomal gene unless these phages contain S. coelicolor chromosomal DNA. In such lysogens, the glk gene becomes integrated into the chromosome by homologous recombination directed by the chromosomal insert on the phage DNA. In appropriate selective conditions, mutants which contain deletions of the glk gene that extend into the adjacent host DNA can be easily isolated. This method was used to insert glk into the methylenomycin biosynthetic genes, and isolate derivatives with deletions of host DNA from within the prophage into the adjacent host DNA. Phenotypic and Southern blot analysis of the deletions showed that there are no genes essential for methylenomycin biosynthesis for at least 13 kb to the left of a region concerned with negative regulation of methylenomycin biosynthesis. Many of the deletions also removed part of the phi C31 prophage. PMID- 3033440 TI - The yeast nuclear gene CBS1 is required for translation of mitochondrial mRNAs bearing the cob 5' untranslated leader. AB - Mitochondrial translation of the cob mRNA to yield apocytochrome b is specifically dependent on the nuclear gene CBS1, while mitochondrial translation of the oxi2 mRNA to yield cytochrome oxidase subunit III (cox III) is specifically dependent on the nuclear gene PET494. Chimeric oxi2 mRNAs bearing the 5' leaders of other mitochondrial mRNAs, transcribed from rho- mitochondrial DNAs termed MSU494, are translated in pet494 mutants. In this study, we examined translation of coxIII from MSU494-encoded chimeric mRNAs in zygotes of defined nuclear and mitochondrial genotype. CoxIII was translated from a chimeric mRNA bearing the cob leader only when the zygotes contained a wild-type CBS1 gene. CoxIII translation from an mRNA bearing the 5' leader of the mitochondrial gene aap1 was not dependent on CBS1 activity. We conclude that the product of the nuclear gene CBS1, or something under its control, acts in the mitochondrion on the cob mRNA 5' leader to activate translation of down-stream coding sequences. PMID- 3033441 TI - Overproduction of DnaA protein stimulates initiation of chromosome and minichromosome replication in Escherichia coli. AB - Increased synthesis of DnaA protein, obtained with plasmids carrying the dnaA gene controlled by the heat inducible lambda pL promoter, stimulated initiation of replication from oriC about threefold. The overinitiation was determined both as an increase in copy number of a minichromosome and as an increase in chromosomal gene dosage of oriC proximal DNA. The additional replication forks which were initiated on the chromosome did not lead to an overall increase in DNA content. DNA/DNA hybridization showed an amplification encompassing less than a few hundred kilobases on each side of oriC. Kinetic studies showed that the overinitiation occurred very rapidly after the induction, and that the initiation frequency then decreased to a near normal frequency per oriC. The results indicate that the DnaA protein is one important factor in regulation of initiation of DNA replication from oriC. PMID- 3033442 TI - Molecular cloning and nucleotide sequence of the mutT mutator of Escherichia coli that causes A:T to C:G transversion. AB - The Escherichia coli mutator gene mutT, which causes A:T----C:G transversion, was cloned in pBR 322. mutT+ plasmids carry a 0.9 kb PvuII DNA fragment derived from the E. coli chromosome. Specific labelling of plasmid-encoded proteins by the maxicell method revealed that mutT codes for a polypeptide of about 15,000 daltons. The protein was overproduced when the mutT gene was placed under the control of the lac regulatory region on a multicopy runaway plasmid. The nucleotide sequence of the mutT gene was determined by the dideoxy method. PMID- 3033443 TI - Reduced transcription of the rnh gene in Escherichia coli mutants expressing the SOS regulon constitutively. AB - We have analysed the transcription levels for the convergently overlapping Escherichia coli genes for the DNA polymerase III proofreading function (dnaQ) and ribonuclease H (rnh). The two tandem dnaQ promoters are about three times more active than the single rnh promoter as shown by analysing the level of in vivo transcription using dnaQ-galK and rnh-galK fusions. In E. coli mutants constitutively expressing the pleiotropic SOS response, which includes activities that enhance DNA repair, recombination and mutagenesis, a strong reduction in rnh transcription was observed. The lexA51 recA441 double mutant which fully expresses the SOS response shows the strongest reduction in rnh transcription and the highest increase in dnaQ transcription. Nuclease S1 mapping supported the finding that a constitutive expression of SOS function leads to a strong reduction in rnh transcription. PMID- 3033445 TI - Characterization of the nuclear progesterone receptor in normal Premarin-primed post-menopausal endometrium. AB - Nuclear progesterone receptors (RPN) were characterised in a 0.4 M KCl nuclear extract of Premarin-primed normal post-menopausal human endometrium after progesterone injection using [3H]R5020 as a ligand. The receptor character of the binding was demonstrated by the following findings: the high specificity for progesterone binding; the high saturable affinity (dissociation constant approx. 2.5 nM) for R5020; the sedimentation coefficient at high salt concentration (approx. 3 S); and the change in distribution of the RP following progesterone administration. These results indicate that normal post-menopausal human endometrium retains the property of translocation (2-step model) or nuclear activation and retention (nuclear receptor model) of the RP if it is exposed to the appropriate oestrogenic and progestational hormonal milieu. PMID- 3033444 TI - Benefits and risks of different hormonal replacement therapies in post-menopausal women. AB - A total of 113 women who presented with climacteric symptoms participated in the study. They were randomly allocated to seven groups of 10-27 subjects, who received for 6 mth the following therapies, respectively: conjugated oestrogens (CE) 0.625 mg/day for 21 days + norethisterone (NET) 5 mg/day from day 12 to day 21; CE + cyproterone acetate (CPA) 12.5 mg/day from day 1 to day 10; oestradiol valerate (EV) 2 mg/day for 21 days + NET; EV + CPA; oestriol (E3) 2-4 mg/day; tibolone (ORG OD14) 2.5 mg/day; and placebo, one tablet/day. Hot flushes decreased significantly over the treatment period in all seven groups. However, E3 was less effective at the dose used than CE, EV or ORG OD 14. At the end of the 6 month treatment period histological examination revealed no changes in endometrial morphology in any of the patients treated. Indeed, the addition of a progestogen even induced regression of endometrial hyperplasia in 8 cases. No significant variation in the plasma levels of triglycerides, total cholesterol, high-density lipoprotein (HDL) or low-density lipoprotein (LDL) was observed after the second and sixth months of treatment with E3 or ORG OD 14. After 6 months, treatment with CE/EV + CPA produced a significant increase in HDL, while treatment with CE/EV + NET brought about a reduction in total cholesterol and HDL and an increase in LDL. PMID- 3033446 TI - Effect of progesterone injection on progesterone receptor levels and distribution in untreated and short-term Premarin-primed pre-menopausal and post-menopausal endometrium. AB - Cytosolic and nuclear progesterone receptors (RPC, RPN) were measured in post menopausal endometria, using [3H]R5020 as the radioligand, and the findings compared with those in pre-menopausal endometria. Total RP levels (RPC + RPN) in post-menopausal endometria were low, i.e. less than 2000 fmol/mg DNA. A 7-11 day course of Premarin (conjugated equine oestrogen) treatment in post-menopausal subjects resulted in RP levels in 11818 +/- 3008 fmol/mg DNA, which were higher than those in proliferative, mid-cycle and Premarin-primed pre-menopausal endometria. Progesterone injection 1-3 h before tissue collection resulted in a change in the distribution of the RP in both premenopausal and post-menopausal Premarin-primed endometria and pre-menopausal proliferative and mid-cycle endometria. Following the progesterone injection RPN levels increased to 57 +/- 9% of the total as compared with 23 +/- 8% in endometrial samples from women who received no progesterone. PMID- 3033448 TI - Comparison of antibody response in mice to Sendai virus exposed to disulfide bonds splitting or U.V. irradiation. AB - The effect of treating Sendai Virus with reducing agents as dithiothreitol (DTT) or with ultraviolet (U.V.)--irradiation, upon the induction of an immune response in Balb/c mice, was studied. Humoral immune response was assessed measuring the haemolysis inhibiting (HLI) antibody titre in serum 1 and 2 weeks after injection. The native viral particles elicited an HLI antibody titre proportional to the amount of viral proteins injected in the range of 8-300 micrograms. The U.V.--irradiation of viral inoculum produced a 6-7 fold decrease in HLI antibody titre even raising the injected dose to 740 micrograms. Similarly, dithiothreitol treatment, which abolishes binding and fusogenic properties of Sendai virus particles, affected the HLI antibody titre about 2-6 fold. PMID- 3033447 TI - Conservation and map location of human cytomegalovirus strain Ad-169 transforming sequences in the DNA of clinical isolates. AB - All DNAs from human cytomegalovirus studied (57 fresh isolates, 3 reference strains) showed hybridization under stringent conditions to a plasmid containing strain Ad-169 transforming sequences. Analysis of hybridization patterns obtained with DNAs digested by three different enzymes allowed construction of a global map for this highly conserved area of the genome. PMID- 3033450 TI - Reactivation in calves of latent infection by Bovid herpesvirus-4. AB - Nine calves, six of which had been infected with strain 85/BH 16TV and three with strain 85/BH 232TN of Bovid herpesvirus-4 (BHV-4), were treated with dexamethasone (DMS) three months after infection. DMS administration did not induce any clinical signs of disease, but BHV-4 was isolated from the nasal swabbings of all calves for a maximum of 8 days after the start of DMS treatment. The virus was also isolated from the nerve tissues, nasal mucosa, lymph nodes, lung and spleen of 4 calves that were killed at different stages (3, 5, 7 days) of the DMS treatment. Intranuclear inclusions associated with cellular shrinkage were found in the neural tissues of calves killed either 3 or 5 days after the start of DMS treatment. Unexpectedly, a latent infection by infectious bovine rhinotracheitis (IBR) virus was reactivated. The virus was isolated from the nasal swabbings of two calves and also from the brain, cerebellum and nasal mucosa of one calf killed 5 days after the start of DMS treatment. PMID- 3033449 TI - Development of immunity to Epstein-Barr virus in Malaysian children. AB - The pattern of seroconversion to Epstein-Barr virus (EBV) was determined in 98 Malaysian children aged 2 weeks to 12 years. Maternal IgG antibodies to EBV viral capsid antigen, ranging between 1:10 to 1:160 titer, were found in 70.6 percent of infants less than three months old, and dropped to 26 percent by seven to nine months. Primary infection, as denoted by emergence of EBV-IgM antibody, occurred at 4 to 6 months, and by eight years all children were seropositive. Maternal antibody titers to EBV nuclear antigen were detected in 52.9 percent of infants less than 3 months old, declined to undetectable levels by 4 to 12 months, and then increased to 40 percent by the age of 12 years. The IgA antibody to viral capsid antigen was absent in all but one infant aged one year; the child also had IgG anti-early antigen, The IgG antibody to EBV early antigen were present in 17.7 percent of the infants aged 3 months or less. This seroconversion to EBV in early life explains the absence of infectious mononucleosis in the Malaysian population. The data suggest that a subunit vaccine to protect against EBV associated diseases, most notably nasopharyngeal carcinoma, commonly observed in Malaysians would have to be administered to infants 6-12 months of age. PMID- 3033451 TI - Interferon-gamma inhibits cell replication, but not pp60src activity of RSV transformed fibroblasts. AB - The present study analyzes the effect of murine IFN-gamma on DNA synthesis, ornithine decarboxylase activity and phosphorylation of pp60src of Rouse sarcoma virus transformed cells. When natural or recombinant IFN-gamma was added to quiescent SR-BALB or L1210 cells, stimulated with fetal calf serum, only the highest IFN-gamma concentrations (2000 U/ml) inhibited DNA synthesis of both tumor lines. By contrast, lower IFN-concentrations (20-200 U/ml) inhibited the DNA synthesis of L1210 but not of SR-BALB tumor cells. A similar pattern of inhibition was observed when ornithine decarboxylase activity was analyzed. Finally, when the levels of phosphorylation in SR-BALB cells were analyzed after IFN-gamma treatment, no difference with untreated controls was observed, even at the highest concentrations. These results suggest that SR-BALB tumor cells are insensitive to IFN-gamma and that src oncogene activity is not affected at IFN concentrations inhibiting cell growth. PMID- 3033452 TI - Effects of lipid A content, hydrocortisone and polymyxin B on endotoxin-induced decreases in in vivo drug metabolism. AB - Using hexobarbital sleeping and zoxazolamine paralysis time as indices of in vivo hepatic drug metabolism, the effects of endotoxin on drug action appear to be time- and dose-dependent and the lipid A moiety of endotoxin appears to be responsible for its inhibitory effects. These studies have also demonstrated that polymyxin B can ameliorate the adverse effects of endotoxin on drug metabolism. Since hydrocortisone protects mice from endotoxin lethality, but does not alter the prolongation of hexobarbital sleeping time caused by endotoxin, it is possible to separate the lethal effects of endotoxin from its effects on drug metabolism. PMID- 3033454 TI - An unusual case of childhood paraplegia. PMID- 3033453 TI - Leukaemogenesis: a postulated mechanism involving tyrosine protein kinase and DNA topoisomerase. AB - The weight of available evidence suggests that leukaemogenesis is a multi-step, complex process. Since there are different types of leukaemia, it seems likely that a variety of distinct molecular mechanisms are involved, arising from different combinations of genetic defects. It is not therefore surprising that studies at genetic, karyotypic, biochemical, cellular and clinical levels all reveal considerable heterogeneity of abnormalities. An attempt to define associations between abnormalities at these different levels in the myeloid leukaemias, led to a new hypothesis which provides a link between activation of the tyrosine kinase group of oncogenes and two components of multi-step leukaemogenesis: reduced differentiation and the tendency for acquisition of further genetic changes. PMID- 3033455 TI - Use of sodium restriction and enalapril in persons with moderate to severe hypertension. AB - One hundred and seventy-four patients who were receiving drug therapy for hypertension were asked to restrict their sodium intake for three months. At the end of that time their drug therapy was replaced with enalapril and the dose of the drug "titrated" to obtain a diastolic blood pressure of less than 90 mmHg. Sodium restriction caused a small fall in blood pressure and could be used as sole therapy in only 6% of patients. Enalapril therapy was instituted without problems and control of blood pressure below 90 mmHg was achieved in 62% of persons with monotherapy. The number of tablets of enalapril that were taken was reduced from 5.9 to 2.7; in most patients these were taken once a day. There were few side-effects and no depression of white cell count, no proteinuria and no deterioration of renal function. Seventy-six per cent of patients preferred the new regimen either because they felt better than with their previous therapy (52%) or because of the more simple regimen (24%). Enalapril was an effective, well tolerated antihypertensive agent and potentially has a major role to play in the management of patients with high blood pressure. PMID- 3033456 TI - Molecular weight determination of the two segments of double-stranded RNA of infectious bursal disease virus, a member of the birnavirus group. AB - The molecular weights (mol. wts.) of the two double-stranded (ds) RNA segments of infectious bursal disease virus (IBDV) were determined using previously sequenced reovirus genes M3 and S2 as internal ds RNA reference molecules. Electrophoresis under fully denaturing conditions revealed mol. wts. of 2.26 X 10(6) daltons and 1.98 X 10(6) daltons. By direct length measurements under the electron microscope, using two different spreading conditions, the two segments were calculated to be composed of 3274 +/- 79 base pairs (bp) and 2821 +/- 59 bp or 3299 +/- 68 bp and 2830 +/- 73 bp, resulting in mol. wts. of 2.24-2.26 X 10(6) daltons and 1.93-1.94 X 10(6) daltons, respectively. Base pair distances of 2.67 +/- 0.08 A and 2.71 +/- 0.11 A in ds RNA were close to those of the A-RNA form; in ds DNA included as a control, the rise per base pair was 3.18 A, which is consistent with published results. Mol. wts. obtained for IBDV indicate that the RNAs of the other birnaviruses are also smaller than reported. PMID- 3033457 TI - Effect of trypsin and chymotrypsin on polypeptides of human rotavirus KUN strain. AB - In view of the fact that trypsin enhances the infectivity of human rotavirus and decreases its hemagglutination, the trypsin-mediated structural modification of the viral polypeptides was analysed, using the KUN strain, a cultivable human rotavirus isolate, grown in the absence of trypsin. A major polypeptide sensitive to trypsin treatment was Vp4 with a molecular weight of 80,000, being cleaved into three polypeptides with molecular weights of 54,000 (P54), 30,000 (P30), and 24,000 (P24). Vp4 was also sensitive to chymotrypsin treatment, generating cleavage products different from those obtained with trypsin. PMID- 3033458 TI - The regulated expression of Epstein-Barr virus: evidence that the transition from primary to secondary protein synthesis in Raji cells superinfected with Epstein Barr virus requires the synthesis of new RNA. AB - In the presence of canavanine and the absence of arginine, superinfected Raji cells synthesize a limited spectrum of proteins (primary proteins). After the removal of canavanine at 8 h post infection, the cells proceed after a lag phase of 2-3 h to synthesize a second group of proteins. The appearance of these proteins can be prevented by addition of actinomycin-D to the chase media, suggesting that active primary proteins are required for the synthesis of new mRNA coding for the secondary proteins. PMID- 3033459 TI - [Pigmented neuroectodermal tumor in childhood. Description of a case]. PMID- 3033460 TI - How research convinced my hospital to start a new cardiac rehab program. PMID- 3033461 TI - Two nurse groups marshall efforts to fight unfair lay-offs, save jobs. Part III. PMID- 3033462 TI - B-virus infection in humans--Pensacola, Florida. PMID- 3033463 TI - [Evaluation of the prognostic factor in human gastric cancer with special reference to the relationship between proliferation and carcinoembryonic antigen production of the cancer cell]. AB - The biological property of human gastric cancer was studied by morphological and immunohistochemical observations. The cytoplasmic mucin of cancer cells was mainly neurtromucin in the mucosa. However, acid mucin was increased in the cancer cells of signet-ring cell carcinoma with submucosal infiltration. Immunohistochemically, CEA was localized in the plasma membrane of matured signet ring cells and the cytoplasm of immature cancer cells. On electron microscopic observation, the cytoplasmic organellae were prominent in the immature cells which were dominant in submucosal infiltrating region. In vitro, the cancer cell line established from signet-ring cell carcinoma revealed that CEA production was associated with cell growth property showing 3H-TdR and BrdU uptake. This results may suggest that CEA production was an important indicator for the decision of prognosis in signet-ring cell carcinoma. PMID- 3033465 TI - Stereochemical features controlling binding and intrinsic activity properties of benzodiazepine-receptor ligands. AB - Benzodiazepine-receptor ligands belong to several different chemical classes. All of them bind to the receptor but display a variety of biological effects ranging from agonist to inverse agonist to antagonist. The properties of the most representative compounds for each class are briefly reviewed as concerns their receptor binding affinities, gamma-aminobutyric acid ratios, photoaffinity labeling ratios, and pharmacological properties. Their geometries, as obtained by X-ray crystallography, are discussed and missing crystal and molecular structures of two of them (zopiclone and CL 218-872) are reported. Binding and intrinsic activity properties of series of benzodiazepines and beta-carbolines are extensively analyzed and correlated with their molecular structures. A general stereochemical model accounting for both binding abilities and kinds of biochemical and pharmacological activities for all benzodiazepine-receptor ligands is proposed. This is based on the assumption of a rather diffuse and substantially planar recognition site where the main drug-receptor interactions are mediated by the drug carbonylic or iminic groups via hydrogen bonding and the observed differences in pharmacological profiles are accounted for by the different localization of the different ligands inside this unique binding site. PMID- 3033464 TI - Characterization of the binding of a morphine (mu) receptor-specific ligand: Tyr Pro-NMePhe-D-Pro-NH2, [3H]-PL17. AB - The highly mu-receptor-selective ligand Tyr-Pro-NMePhe-D-Pro-NH2 (PL17) was custom tritiated and its binding to rat brain membranes was directly measured. The data were well fit by a model assuming the existence of a single homogeneous population of binding sites. Scatchard analysis yielded values for Kd and binding sites of 6 nM and 0.16 pmol/mg, respectively. As expected for binding to an opiate receptor, addition of sodium, magnesium, and guanyl nucleotides to the assays resulted in a modulation of ligand binding. In all cases tested, however, no significant deviations of the Scatchard plots from linearity were observed. Furthermore, displacement of [3H]-PL17 by all opioid ligands tested was monophasic, consistent with simple competitive inhibition at a single binding site. The IC50 values thus obtained showed good agreement with previously published values determined using less selective radiolabeled ligands. The regional distribution of [3H]-PL17 binding to rat brain was examined by in vitro autoradiography. The labeling pattern was similar to that seen for other mu-type ligands. Because of its high selectivity, the binding of [3H]-PL17 to the mu-type receptor can be measured directly without the need to suppress binding to other sites. As such, [3H]-PL17 should be a useful ligand for dissecting the actions of multiple opiate receptors. PMID- 3033466 TI - Glucocorticoid regulation of beta-adrenergic receptors in 3T3-L1 preadipocytes. AB - Treatment of 3T3-L1 preadipocytes (fibroblasts) with 250 nM dexamethasone for 48 hr caused a doubling of total beta-adrenergic receptors and an increase in beta 2 adrenergic receptor subtype proportion from approximately 50% in controls to 85% in treated cells. The responses to epinephrine and norepinephrine in a whole cell cAMP accumulation assay reflected these changes. The effects of dexamethasone on beta-adrenergic receptors were mediated through the glucocorticoid receptor and were time and dose dependent with an EC50 of 2.77 +/- 0.73 nM for an increase in the proportion of beta 2-adrenergic receptors. The rank order of potency of steroids to effect these changes (betamethasone = dexamethasone greater than fludrocortisone greater than hydrocortisone = triamcinolone greater than aldosterone) correlated with their glucocorticoid potency. [3H]Dexamethasone binding to intact cells yielded a KD value of 3.47 +/- 0.38 nM for binding to the glucocorticoid receptor which correlated well with the EC50 for dexamethasone to alter beta-adrenergic receptors. Inhibition of [3H]dexamethasone binding by other steroids confirmed that the ability of steroids to regulate beta-adrenergic receptors correlated with the affinity of each compound for the 3T3-L1 glucocorticoid receptor. Progesterone, which can bind to the glucocorticoid receptor but has only weak agonist activity, competitively inhibited the ability of dexamethasone to alter beta-adrenergic receptors. Protein synthesis, RNA synthesis, and N-linked glycosylation appeared to be necessary for the change in receptor subtype expression and the increase in beta-adrenergic receptor number induced by dexamethasone. The present study suggests that regulation of beta adrenergic receptor expression in 3T3-L1 preadipocytes by dexamethasone is a glucocorticoid-specific effect which may require gene activation. PMID- 3033467 TI - Oxidation of thiol drugs and biochemicals by the lactoperoxidase/hydrogen peroxide system. AB - The thiol moiety is prone to oxidative free radical formation, which may be important in mediating the toxicity of some thiol-containing compounds. The oxidation of the compounds cysteine, cysteamine, N-acetylcysteine, glutathione, penicillamine, and captopril were studied using ESR and oxygen uptake techniques. Lactoperoxidase, with hydrogen peroxide to provide oxidizing equivalents, was used to initiate the oxidation. The reaction appears to be strongly peroxide dependent, with either exogenous H2O2 or thiol-derived peroxide driving the reaction. PMID- 3033468 TI - Incorporation of (E)-5-(2-iodovinyl)-2'-deoxyuridine into deoxyribonucleic acids of varicella-zoster virus (TK+ and TK- strains)-infected cells. AB - The incorporation of (E)-5-(2-iodovinyl)-2'-deoxyuridine (IVDU) into DNA of varicella-zoster virus (VZV)-infected human embryo fibroblasts was studied, using thymidine kinase-positive (TK+) and thymidine kinase-negative (TK-) VZV strains. [125I]IVDU was taken up by cells infected with TK+ VZV-, but not by TK- VZV- or mock-infected cells. [125I]IVDU was incorporated into both VZV DNA and cellular DNA of TK+ VZV-infected cells. When the cells were exposed to 0.3 microM IVDU, a more marked shift was noted in the buoyant density of viral DNA than of host DNA. In contrast, the DNAs isolated from TK- VZV- or mock-infected cells did not exhibit a detectable incorporation of [125I]IVDU. [125I] IVDU-labeled VZV DNA was purified from the viral nucleocapsids of TK+ VZV-infected cells. Substitution of no more than 0.1-1% of the thymidine residues in the VZV DNA by IVDU seemed to suffice to inhibit the replication of VZV. PMID- 3033469 TI - CGS 9343B, a novel, potent, and selective inhibitor of calmodulin activity. AB - 1,3-Dihydro-1-[1-[(4-methyl-4H,6H-pyrrolo[1,2-a][4,1]- benzoxazepin-4-yl)methyl] 4-piperidinyl]-2H-benzimidazol-2-o ne (1:1) maleate was synthesized in six steps from methyl anthranilate and designated CGS 9343B. CGS 9343B inhibited calmodulin stimulated cAMP phosphodiesterase activity with an IC50 value of 3.3 microM. CGS 9343B was 3.8 times more potent than trifluoperazine (IC50 = 12.7 microM) as an inhibitor of calmodulin activity. CGS 9343B did not inhibit protein kinase C activity at concentrations up to 100 microM, whereas trifluoperazine inhibited protein kinase C activity with an IC50 value of 43.9 microM. CGS 9343B weakly displaced [3H]spiperone from postsynaptic dopamine receptors with an IC50 value of 4.8 microM while the value for trifluoperazine, a potent antipsychotic agent, was 0.018 microM. It is concluded that CGS 9343B is a novel, potent, and selective inhibitor of calmodulin activity. Unlike trifluoperazine, CGS 9343B does not inhibit protein kinase C activity and does not possess potential antidopaminergic activity. PMID- 3033470 TI - Repeated electroconvulsive shock: effect on sodium dependency and regional distribution of opioid-binding sites. AB - The effects of single and repeated electroconvulsive shock (ECS) on the binding of [3H]diprenorphine to rat brain membranes was studied. Repeated but not single ECS significantly increased the Bmax of [3H]diprenorphine binding when measured in the absence but not in the presence of NaCl. On a regional basis the effect of ECS was greatest in the olfactory bulb, nucleus accumbens, and striatum. More modest increases were found in the hippocampus, amygdala, septum, hypothalamus, and pyriform cortex. No significant effect was found in the brainstem and frontal cortex. Although the regional rank order of receptor increase does not match the receptor distribution of brain enkephalins, the receptor increase does parallel the regional increases in brain enkephalins following ECS. PMID- 3033471 TI - [Molecular mechanisms of photoreception. VI. Cyclic nucleotide- and light dependent phosphorylation of rod outer segment proteins in the frog retina]. AB - Phosphorylation of proteins in purified rod outer segment from frog retina was investigated. Phosphorylation of 18, 17, 12 and 11.5 kDa proteins was stimulated by cAMP (Ka approximately equal to 10(-7) M) and cGMP (Ka approximately equal to 10(-4) M). 32P-incorporation into 18 and 17 kDa proteins was much lower than into 12 and 11.5 kDa, which are in the group of main phosphoproteins of the rod outer segment: 12 and 11.5 kDA phosphoproteins appear to be present in cytoplasm or are slightly bound to disk's membranes. However, they are not discovered in the cytoplasmic membranes. The dephosphorylation of low-molecular weight proteins, discovered earlier by Polans et al., occurs slowly: the light doesn't change the level of phosphorylation of proteins in living retina within the time of photoresponse. It is suggested that the process of light-dependent phosphorylation-dephosphorylation of 12 and 11.5 kDa proteins controls the light sensitivity of the photoreceptor. PMID- 3033473 TI - [Plasmid vector for cloning, determination of nucleotide sequence and directed assembly of DNA fragments]. AB - A plasmid vector pNIMB has been constructed (starting) from the pUR222 plasmid as a result of substitution of the polylinker containing restriction sites: PstI, SalGI, AccI, HindII, BamHI EcoRI and by other synthetic linkers with additional sites for HindIII and HgaI. Plasmid pNIMB does not differ from the parent one phenotypically. Compared to pUR222 the vector contains an additional site for cloning HindIII fragments of DNA and allows to clone SalGI/BamHI- and PstI/SalGI fragments. Cloning of DNA fragments in all seven unique sites of pNiMB gives the possibility for sequencing the fragments avoiding their isolation from the gel. Moreover, this vector may be useful for cloning and directed assembly of chemically synthesised DNA fragments when the endonuclease HgaI sites are used. PMID- 3033472 TI - [Quantum chemistry analysis of the mechanism of action of proteolytic enzymes. III. Proton transport in serine proteases]. AB - The model system for the proton transfer on the amide atom of the substrate leaving group based on the existence of "charge relay system" in the serine type proteases was analysed by the CNDO/2 method. The unfitness of this model to explain the action mechanism of serine proteases was shown. The model system for proton transfer with the water molecule as the intermediate acceptor of the Ser 195 proton was suggested and analysed by the same method. The acylation activation barrier of this system was shown to localize on the stage of synchronous transfer of the Ser-195 alcoholic proton and the water molecule proton hydrogen bound to the His-57 N epsilon 2-atom on the water molecule oxygen atom and the N epsilon 2-atom, respectively. The protonation of substrate in the case of the model system with the water molecule as the intermediate acceptor of proton was demonstrated to begin before the completion of the tetrahedral intermediate substance and the protonated from of the tetrahedral intermediate was shown to form only. A hypothesis considering the role of this water molecule as the nucleophilic reagent on the deacylation stage is presented. PMID- 3033474 TI - [Minor promoters of phage phi X174 are controlled by CRP-cAMP, lexA, glnG and several other common common regulatory systems of the host cell]. AB - It was found that CRP-cAMP-recognized sequences in DNA being suggested as GTGN7 11CAC (with variability both in domain's structures and in spacer's length) are located non-randomly in promoters. In CRP-cAMP-stimulated promoters they lie upstream the "-35" box and are separated from it by a whole number of DNA turns, whereas in CRP-cAMP-repressed ones they are located downstream "-35" in a half whole-turn-number distance. Several CRP-, SOS- and NR1-sites in the phi X174 DNA sequence were found and a few new promoters were deduced from it. PCRP1 lies within gene F and has both CRR and ntrC sites and one SOS-operator, PCRP3 (in gene A) has a CRP site which overlaps with the SOS-operator, PA and PCRP2 (in gene G) have sCRP and PD has a stringent discriminator. Four promotors, PCRP1, PCRP2, PA and PB are cloned in the pBR322 plasmid. For cloned PCRP1 the activation by exogenous cAMP and the SOS-induction by the mitomycin C were observed in vivo in pVYB215-containing cells by increasing the levels of beta lactamase up to 27-fold. The new gene L of the phi X174 is deduced from the DNA sequence. It has two start points, overlaps the gene F inside it and codes for peptides 23 or 19 amino acids in length. These lethal peptides have strong homology in sequence to the cellular protein sulA(sfiA) of E. coli, and L* can cause observed filamentation and death of pVYB215- bearing cells after PCRP1 induction. In the A and A* protein sequences two domains "helix-turn-helix" were found that are homologous to those in CRP and repressors; this makes possible the competition between A* and CRT for its DNA sites that also have some homology. The model of the phi X174 infection cycle control and mechanisms of DNA recognition by CRP-CAMP are discussed. PCRP1 is the first promotor controlled by both three global regulons of E. coli cell. PMID- 3033476 TI - [Fructose-1,6-diphosphatase deficiency. Clinical aspects and diagnosis based on a case report]. AB - In a 2-year-old boy the enzyme defect of fructose-1,6-diphosphatase deficiency could be demonstrated in liver tissue, jejunal mucosa and leukocytes. During the neonatal period the boy had suffered from transient metabolic acidosis and hypoglycemia. At the age of 2 years, during a febrile infection, he developed a hyperkinetic-hypotonic syndrome, which disappeared by fructose-free diet and avoidance of prolonged periods of fasting. PMID- 3033475 TI - [Primary structure of cDNA for bovine beta-casein]. AB - In this work the complete sequence of 1097 nucleotide bovine beta-casein cDNA has been determined. Sequencing of end labeled fragments was performed using the method of Maxam and Gilbert. Bovine beta-casein cDNA consist of a 672 nucleotide coding region, flanked by 60 nucleotide 5' and 358 nucleotide 3'-noncoding region. The restriction map of beta-casein cDNA was constructed. The computer analysis was done and the comparisons of the complete sequence of bovine beta casein cDNA with other sequences, coding caseins in bovine, rat and guinea-pig was performed. It was determined, that cloned cDNA is related to genetic variant A1. PMID- 3033478 TI - Retroperitoneal malignant fibrous histiocytoma: case report and literature review. PMID- 3033477 TI - Assessment of serum angiotensin-converting enzyme as a marker of activity in sarcoidosis: a study of 31 patients with erythema nodosum. PMID- 3033479 TI - Multiplicity of causes for adrenal damage in AIDS. PMID- 3033480 TI - [Mobilization of the pACYC184 plasmid by hybrid pAS8-121 delta plasmids in Escherichia coli cells]. AB - The plasmid pACYC184 is shown to be mobilized for conjugal transfer in Escherichia coli cells by the deleted (Tn7-TcR) derivatives of the hybrid conjugative plasmid pAS8-121 (RP4-Co1E1). Both the mobilized and mobilizing plasmids are autonomously inherited by the recipient cells when the mobilizing plasmid carries single copy of IS8 (the plasmid pAS8-121 delta 16). Cointegrates pAS8-121 delta 16D:: ::pACYC184 are found in the recipient cells with pACYC184 being inserted between two repeats of IS8 if the derivate plasmid pAS8-121 delta 16D having the duplication of IS8 is used to mobilize pACYC184 for conjugal transfer. The insertion of pACYC184 between IS8 repeats in the plasmid pAS8-121 delta 16D eliminates the plasmid ability to be inserted with high frequency into the chromosome of the phototrophic bacterium R. sphaeroides 2R. The cointegrate pAS8-121 delta 16D:: pACYC184 is stable but can be resolved during the transformation deriving the plasmid pACYC184:: IS8. The latter may be used as a probe for isolation and analysis of IS8 DNA sequences and for constructing the vectors on the basis of pACYC184. PMID- 3033481 TI - [Protein Z is not an endonuclease of the recF recombination pathway in Escherichia coli K12]. AB - A 74 kD protein was extracted from Escherichia coli cells and purified under the physiological conditions. The protein is able to catalyze the reactions of endonucleolytic degradation of plasmid DNA. The genetic determinant coding for the 74 KD protein synthesis has been localized between 17 and 27 min on Escherichia coli chromosomal map. The endonuclease previously described as a recF gene dependent "protein Z" (Krivonogov S. V., Novitskaja V. A. Mol. Gen. Genet., 1982, v, 187, p. 302) is shown to be independent of the integrity of Escherichia coli recF gene. PMID- 3033482 TI - [Properties of hepatitis A virus particles produced during infection in vitro]. AB - Four types of virus-specific particles with different sedimentation coefficients and buoyant densities in CsCl were shown to be accumulated in hepatitis A virus (strain HAS-15) infected fetal rhesus monkey kidney cells (FRhK-4 line). Unlike the mature virions (155S, 1.34 g/cm3), cell-associated isosedimenting 92 S particles (buoyant densities of 1.30 and 1.20 g/cm3) proved to be sensitive to lipase action. Particles of all four types were shown to contain similar sets of polypeptides, and, with the exception of "empty" 1.30 g/cm3-particles, appeared to be "full" under the immune electron microscopic examination. The viral RNA was unequivocally identified by the molecular hydridization test only in the mature virions. PMID- 3033483 TI - [Detailed mapping of the Hly-region of the plasmid pHly241. Homology of the hemolysis determinants of plasmids pHly241 and pHly195]. AB - Data on the hemolysin synthesis determined by the plasmid pHly241, belonging to the incompatibility group I2, and its derivatives are presented in this paper. The restriction analysis of the pHly241 plasmid has resulted in the detailed mapping of the hly determinant region in the plasmid. The substantial homology has been demonstrated between the regions of the plasmids pHly241 and pHly195 coding for hemolysin synthesis and excretion from the bacterial cell. PMID- 3033484 TI - [Primary structure of the potato virus X genome: the region preceding the capsid protein cistron]. AB - DNA copies of the potato virus X (PVX) RNA corresponding to 2300 nucleotides at the 3'-end have been cloned. The cloned cDNA copies containing the nucleotides 445-1280 from the 3'-end have been sequenced. The 5'-terminal region of the PVX coat protein gene corresponds to residues 445-786 from the 3'-end. The amino acid sequences of two more open reading frames (ORF) have been deduced from the nucleotide sequence. The potential translation products of these ORF's would correspond to the nonstructural viral proteins. We have located the ORF1 within the region of residues 799-1009 preceding the coat protein cistron. The tentative protein is composed of 70 amino acids and has an aminoterminal segment which is markedly hydrophobic. ORF2 in the PVX sequence ends with UAG at nucleotides 942 944 and extends to the 5'-terminus for additional 340 nucleotides. The distant sequence homology exists between a carboxyterminal portion of PVX ORF2 and that of the nonstructural "30 K-proteins" of the plant tobamoviruses. PMID- 3033485 TI - [The genome of chickens free of endogenous avian leukosis-sarcoma proviruses contains sequences distantly related to Rous sarcoma virus]. AB - Line of Brown leghorn chickens free of RAV-O-type endogenous proviruses was obtained by selection under blot hybridization control. A set of dispersed sequences distantly related to avian leukosis virus genome was found in DNA of these chickens by means of hybridization in non-stringent conditions. Different restriction fragments were detected by gag, pol and env hybridization probes. PMID- 3033486 TI - recA-dependent and recA-independent N-ethyl-N-nitrosourea mutagenesis at a plasmid-encoded herpes simplex virus thymidine kinase gene in Escherichia coli. AB - We have compared isogenic recA13/recA+ Escherichia coli K-12 strains for the induction by N-ethyl-N-nitrosourea (ENU) of forward mutations at a plasmid encoded herpes simplex virus type 1 thymidine kinase (HSV-tk) gene. Treatment of plasmid-bearing bacteria with ENU resulted in a dose-dependent increase in the mutant frequencies of the chromosomal udk locus and of the plasmid HSV-tk locus in both recA13 and recA+ strains. Although the recA13 strain was considerably more sensitive to the cytotoxic effects of ENU treatment than was the recA+ strain, the ENU-induced mutation frequency at both loci was greater for the recA+ strain than for the recA13 strain. When plasmid DNA modified by in vitro reaction with ENU was used to transform recA13, recA+, and UV pre-irradiated recA+ strains, an increase in the HSV-tk mutant frequency was observed in all 3 cases. The induction of mutations in recA13 and recA+ strains followed a similar dose response, while the ENU-induced HSV-tk mutant frequency was significantly greater for UV pre-irradiated recA+ bacteria. These results indicate that fixation of ENU induced premutagenic lesions can occur by both recA-dependent and recA independent pathways. PMID- 3033487 TI - Molecular and biochemical analyses of spontaneous and X-ray-induced mutants in human lymphoblastoid cells. AB - We have isolated a series of 14 spontaneously arising and 28 X-ray-induced mutants at the hypoxanthine-guanine phosphoribosyltransferase (hgprt) locus in human lymphoblastoid cells. Among the spontaneous mutants, 5/14 (36%) had detectable alterations in their restriction fragment pattern after hybridization with a human cDNA probe for hgprt. Of the 10 remaining mutants, 4 had partial HGPRT enzyme activity, which suggested that they contained point mutations. Among the 28 mutants induced by 150 rad of X-rays, 15 (54%) had deletions of part or all of the hgprt gene. 5 of the remaining 13 (18% overall) had partial HGPRT enzyme activity, which suggested that they contained point mutations. These data imply that in this human cell system, X-rays induce both point mutants which have residual enzyme activity as well as mutations involving relatively large deletions of DNA. PMID- 3033489 TI - Effect of heat treatment on chromosomal aberrations induced by the alkylating agent trenimon or the restriction endonuclease Alu I in Chinese hamster ovary (CHO) cells. AB - Heat treatment of CHO cells in the G1-phase of the cell cycle leads to chromatid type aberrations in first posttreatment metaphases. Posttreatment of heat-treated cells with the alkylating agent trenimon leads to a synergistic effect on the production of chromatid-type exchanges. These results indicate that heat induces lesions which like the lesions produced by trenimon give rise to chromatid-type aberrations during the first posttreatment S-phase, and that these lesions can interact with each other to produce chromatid-type exchanges. Treatment of CHO cells in the G1-phase of the cell cycle with the restriction endonuclease Alu I induces chromosomal aberrations. Pretreatment of cells with heat leads to a reduction of Alu I induced chromosome-type aberrations. When cells are allowed to recover after heat treatment for 22 h, the aberration frequencies produced by Alu I are the same as in cells not treated with heat. These findings can be explained by assuming that heat-induced accumulation of accessory proteins in the chromatin protects the DNA from being cut by Alu I, and that the cells recovered from the heat-induced protein accumulation after 22 h. PMID- 3033488 TI - Iron-mediated induction of sister-chromatid exchanges by hydrogen peroxide and superoxide anion. AB - When Chinese hamster fibroblasts were exposed to hydrogen peroxide or to a system consisting of xanthine oxidase and hypoxanthine, which generates superoxide anion plus hydrogen peroxide, sister-chromatid exchanges (SCEs) were formed in a dose dependent manner. When the iron-complexing agent o-phenanthroline was present in the medium, however, the production of these SCEs was completely inhibited. This fact indicates that the Fenton reaction: Fe2+ + H2O2----OH0 + OH- + Fe3+ is responsible for the production of SCEs. When O2- and H2O2 were generated inside the cell by incubation with menadione, the production of SCE was prevented by co incubation with copper diisopropylsalicylate, a superoxide dismutase mimetic agent. The most likely role of O2- is as a reducing agent of Fe3+: O2- + Fe3+--- Fe2+ + O2, so that the sum of this and the Fenton reaction, i.e., the iron catalyzed Haber-Weiss reaction, provides an explanation for the active oxygen species-induced SCE: H2O2 + O2(-)----OH- + OH0 + O2. According to this view, the OH radical thus produced is the agent which ultimately causes SCE. These results are discussed in comparison with other mechanisms previously proposed for induction of SCE by active oxygen species. PMID- 3033490 TI - Nitropyrenes are inducers of polyoma viral DNA synthesis. AB - The biological activity of a series of nitropyrenes was assayed by measuring their ability to induce the asynchronous replication of viral DNA in rat fibroblasts transformed by a ts-a mutant of polyoma virus. Concentrations of 10 30 micrograms/ml of 1-nitropyrene (1-NP) induced viral replication, and this effect was enhanced by addition of rat-liver S9 microsomal fraction (300 micrograms/ml) to the culture medium. The response was less than that obtained with 0.1 micrograms/ml of the activated metabolite of benzo[a]pyrene (BP), BP trans-7,8-dihydrodiol-9,10 epoxide (anti) (BPDE). A series of di-, tri-, and tetra-nitropyrenes were also found to induce polyoma DNA replication, in the absence of exogenous microsomal activation, displaying strongly positive effects at 0.5-2.0 microgram/ml. Dose-response curves with 1,6-dinitropyrene (1,6-DNP) from 0.01 to 0.5 microgram/ml indicated that this compound was approximately equipotent with BPDE for induction of polyoma DNA synthesis. Studies of drug metabolism, DNA binding and DNA adduct formation indicate that 1,6-DNP is metabolized in this cell line, binds to DNA, and forms stable adducts. The level of DNA modification seen with 1,6-DNP is higher than that observed under comparable conditions with an equivalent dose of BPDE. These findings provide additional evidence that the nitropyrene class of compounds can exert biological effects in mammalian cells, and that the dinitropyrenes are more potent than 1 NP. PMID- 3033491 TI - Preirradiation of host (monkey) cells mitigates the effects of UV upon simian virus 40 DNA replication. AB - We are examining the effects of preirradiation of host (monkey) cells upon the replication of UV-damaged SV40. Control cells and cells preirradiated with low fluences (5 or 10 J/m2) of UV were infected with undamaged SV40, and the immediate effects of a subsequent irradiation were determined. UV inhibited total SV 40 DNA synthesis (incorporation of thymidine into viral DNA) in both preirradiated and control cells, but the extent of inhibition was less in the preirradiated cells. A test fluence of 60 J/m2 to SV40 replicating in preirradiated cells reduced synthesis only as much as a test fluence of 25 J/m2 in control cells. The fraction of recently replicated SV40 molecules that re entered the replication pool and subsequently completed one round of replication in the first 2 h after UV was also decreased less in the preirradiated cells. Thus preirradiation of the host cell mitigates the immediate inhibitory effects of a subsequent UV exposure upon SV40 replication. PMID- 3033493 TI - Mutagenic activity of chloramines. AB - Mutagenesis by chloramines and hypochlorous acid (HOCl) was studied to determine whether these agents could contribute to the mutagenic and potentially carcinogenic activity of stimulated leukocytes and whether environmental exposure to these agents is a cause for concern. Mutagenic activity was measured using the S. typhimurium TA97a, TA100 and TA102 tester strains. Because chloramines and HOCl are bactericidal, react rapidly with cell components, and can destroy the histidine and biotin required for the mutagenesis assay, activity can't be compared directly with that of less toxic or reactive agents. Nevertheless, chloramines were mutagenic when tested under appropriate conditions. TA100 was the most sensitive strain, and the most active mutagens were lipophilic dichloramines (RNCl2) including derivatives of histamine, ethanolamine and putrescine. Lipophilic monochloramines (RNHCl) such as histamine-monochloramine and NH2Cl were less active. Hydrophilic chloramines such as taurine-chloramines had low activity, and HOCl was inactive. The metabolic state of the bacteria was critical. Chloramines were mutagenic when added to bacteria with glucose at 37 degrees C, but killing predominated when chloramines were added at 4 degrees C or 25 degrees C, or at 37 degrees C without glucose. Production of chloramines and HOCl by leukocytes in vivo could contribute to the association of chronic inflammation and cancer as a result of: (1) the entry of membrane-permeable chloramines into normal cells followed by attack on intracellular components including DNA, and (2) the production of secondary mutagens such as compounds with carbonyl groups or carbon-chlorine bonds. On the other hand, chlorination of water supplies is perhaps more likely to destroy than create mutagens, and chloramines from the environment are unlikely to penetrate the skin and mucous membranes. PMID- 3033492 TI - Analysis of mutations occurring during replication of a SV40 shuttle vector in mammalian cells. AB - To analyse mutations that arise in mammalian cells we have used a SV40::plasmid shuttle vector containing a portion of the E. coli lacZ gene. We have found that following transfection into monkey Cos-7 cell mutations are not detected in the recovered plasmids at 24 h post transfection, but are found at 48 h post transfection, after the onset of DNA replication. Analysis of the mutant plasmids shows that in almost all cases the mutant phenotype is caused by a deletion or rearrangement of the lacZ gene in the shuttle vector. PMID- 3033494 TI - The ribosomal gene spacer as a tool for the taxonomy of Leishmania. AB - A recombinant DNA ribosomal gene spacer of Leishmania braziliensis Y was used as probe to test different Leishmania species. Based on the similarity of their restriction patterns, three groups were distinguished with respect to international Leishmania references: first a group with a similar restriction pattern to L. braziliensis Y and the reference organism L. mexicana garnhami JAP78; a second group with restriction patterns similar to the reference organism L. mexicana mexicana M379; and finally a group where all the restriction patterns were related to the reference organism L. braziliensis braziliensis M2903. These results support the existence of L. garnhami as an independent Leishmania species; they confirm previous studies on L. mexicana and L. braziliensis and open the way for the more exact diagnosis of New World Leishmaniasis. PMID- 3033495 TI - cDNA clone encoding a high molecular weight antigen of Babesia bovis. AB - An expression library was constructed by inserting cDNA copied from mRNA of the blood stages of Babesia bovis isolate KA into bacteriophage lambda gt11-amp3. An antigen-positive cDNA clone detected by screening the library with antibodies from cattle vaccinated with the KA isolate was shown to encode part of a high molecular weight polypeptide antigen of B. bovis. This molecule was a dominant immunogen and was found by immunofluorescence to be within the parasite in infected erythrocytes. PMID- 3033496 TI - Biochemical changes during the aerobic-anaerobic transition in Ascaris suum larvae. AB - Ascaris suum L3 larvae isolated from rabbit lungs undergo the third ecdysis to L4 larvae after 3 days in culture under a gas phase of 85% N2/10% CO2/5% O2. The L3 larvae contain substantial malic enzyme activity and are capable of producing small amounts of the reduced organic acids characteristic of the fermentative pathways which operate in the adult. However, only a small portion of the total carbon utilized is accounted for by these reduced acids and their motility is cyanide-sensitive, suggesting that their energy-generating pathways are predominantly aerobic. In contrast, after ecdysis, the L4 larvae begin to utilize glucose at a greater rate and the proportion of total carbon utilized which is accounted for as propionate, 2-methylbutyrate and 2-methylvalerate also increases. In addition, motility becomes increasingly cyanide-insensitive, suggesting that these L4 larvae are able to utilize the anaerobic energy generating pathways of the adult. Surprisingly, on day 10 in culture, these L4 larvae, although capable of producing reduced volatile acids, still retain substantial cyanide-sensitive cytochrome oxidase activity. PMID- 3033498 TI - A comparative study of UTP-D-glucose-1-phosphate uridylyl transferase in the cysts of Echinococcus multilocularis and the livers of infected and control Meriones unguiculatus. AB - Kinetic and physical parameters of UDP-glucose pyrophosphorylase were determined in Meriones unguiculatus infected with Echinococcus multilocularis metacestodes (cestoda). Studies were carried out on parasite cysts, and on livers from control and infected animals after purification of the enzyme by affinity chromatography on UTP-agarose. The enzyme from infected and control livers had km values for UTP of 0.01 mM and 0.5 mM, respectively; for glucose-1-phosphate values were 0.46 mM and 0.07 mM, respectively. On the other hand the enzyme from cysts was found to have a higher Km for UTP (1 mM) and for glucose-1-phosphate (1.5 mM) than from infected or non-infected livers. Physical characteristics (pI = 6 and Mr = 160,000) of UDP-glucopyrophosphorylases were the same in controls and infected host livers but were different from the cyst enzyme (pI = 7 and Mr = 251,000). These results provide evidence for the existence of significant differences between parasitic and host enzymes, which could possibly be exploited in chemotherapy. PMID- 3033497 TI - Tandemly arranged gene clusters of malarial parasites that are highly conserved and transcribed. AB - A molecular clone containing a 5.8 kb Eco RI fragment was isolated from a genomic library of the rodent malarial parasite Plasmodium yoelii. The P. yoelii genome contains about 150 copies of this sequence, making up almost 3% of the DNA. These sequences are tandemly arrayed in head-to-tail configurations with the unit length of the repeat being 5.8 kb. Several poly(A+) RNAs of P. yoelii ranging from 1.6 to 0.3 kb are recognized by the 5.8 kb clone. Five additional species of malarial parasites (P. chabaudi, P. berghei, P. falciparum, P. knowlesi, and P. cynomolgi) contain tandemly repeated arrays of sequences having the same unit length of 5.8 kb, which readily hybridize to the sequence cloned from P. yoelii. PMID- 3033500 TI - The effects of p-chloromercuriphenylsulfonic acid on Trypanosoma cruzi infection of mammalian host cells in vitro. AB - Treatment of Trypanosoma cruzi blood trypomastigotes with p chloromercuriphenylsulfonic acid (PCMS) increased the association of the parasite with either mouse resident peritoneal macrophages or rat heart myoblasts in vitro. The effect was evidenced by elevation of both the percentage of host cells with parasites and the number of flagellates per 100 cells. The effect of PCMS appeared to be largely on the process of parasite penetration rather than surface binding as it was seen at 37 degrees C but not at 4 degrees C. A short pretreatment time, 5 min, was sufficient to elicit the enhancement, suggesting that the primary effect of PCMS might be at the parasite's cell surface. The PCMS effect was reversible as the parasite returned to normal levels of association with the host cells in less than 4 h after removal of excess PCMS. That sulfhydryl groups were involved in the PCMS effect was indicated by the abilities of excess cysteine and glutathione to block it. These results suggest a role for free sulfhydryl groups on the parasite surface in the process of host cell invasion. PMID- 3033499 TI - Kinetoplast DNA of Trypanosoma evansi. AB - We show here that the kinetoplast DNA (kDNA) networks from six Trypanosoma evansi strains differ from those of T. brucei by their lack of maxi-circles and absence of mini-circle sequence heterogeneity. The lack of maxi-circles is sufficient to account for the inability of T. evansi to multiply in tsetse flies, since this requires functional mitochondria containing maxi-circle gene products. Judged by restriction enzyme analysis, five of the six T. evansi strains contain mini circles that differ less than 4% in sequence. This type A mini-circle is found in strains from East Africa, West Africa and South America. Another strain from East Africa contains a very different mini-circle (type B), which shows about the same degree of hybridization to type A mini-circles as to a mini-circle from T. brucei. We propose that the pronounced sequence heterogeneity of the mini-circles of T. brucei has arisen by recombination of strains that had diverged for long periods of time in reproductive isolation. We further propose that the homogeneous mini-circles of T. evansi (and T. equiperdum) reflect the inability of species to mate. This proposal implies that mini-circle heterogeneity indicates (infrequent) genetic exchange and that all kinetoplastid flagellates with heterogeneous mini-circles exchange DNA. PMID- 3033501 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 19-1987. A 64-year-old woman with liver dysfunction and abnormal mononuclear cells in the blood smears. PMID- 3033503 TI - Compensating children with vaccine-related injuries. PMID- 3033502 TI - Pituitary tumors containing cholecystokinin. AB - We found small amounts of cholecystokinin in the normal human adenohypophysis and therefore examined pituitary tumors from 87 patients with acromegaly, Cushing's disease, Nelson's syndrome, prolactinoma, or inactive pituitary adenomas. Five adenomas associated with Nelson's syndrome contained increased amounts of cholecystokinin, the concentrations being extremely high in two: 8281 and 13,453 pmol per gram as compared with less than 30 pmol per gram in normal pituitary glands. The cholecystokinin concentrations were moderately increased in adenomas from another 12 patients, of whom 5 had Cushing's disease and 7 acromegaly with adenomas containing ACTH. The cholecystokinin peptides from the tumors were smaller and less sulfated than cholecystokinin from normal pituitary glands. We conclude that ACTH-producing pituitary cells may also produce an altered form of cholecystokinin. PMID- 3033504 TI - Acute hypothalamic-pituitary-adrenal responses to the stress of treadmill exercise. Physiologic adaptations to physical training. AB - To study the effects of physical conditioning on the hypothalamic-pituitary adrenal axis, we examined the plasma ACTH, cortisol, and lactate responses in sedentary subjects, moderately trained runners, and highly trained runners to graded levels of treadmill exercise (50, 70, and 90 percent of maximal oxygen uptake) and to intravenous ovine corticotropin-releasing hormone (1 microgram per kilogram of body weight). Basal evening concentrations of ACTH and cortisol, but not of lactate, were elevated in highly trained runners as compared with sedentary subjects and moderately trained runners. Exercise-stimulated ACTH, cortisol, and lactate responses were similar in all groups and were proportional to the exercise intensity employed. These responses, however, were attenuated in the trained subjects when plotted against applied absolute workload. Only the highly trained group had diminished responses of ACTH and cortisol to ovine corticotropin-releasing hormone, consistent with sustained hypercortisolism. We conclude that physical conditioning is associated with a reduction in pituitary adrenal activation in response to a given workload. Alterations of the hypothalamic-pituitary-adrenal axis consistent with mild hypercortisolism and similar to findings in depression and anorexia nervosa were found only in highly trained runners. Whether these alterations represent an adaptive change to the daily stress of strenuous exercise or a marker of a specific personality profile in highly trained athletes is unknown. PMID- 3033506 TI - Recurrences after oral and genital herpes simplex virus infection. Influence of site of infection and viral type. AB - We prospectively followed 39 adults with concurrent primary herpes simplex virus (HSV) infection (12 with HSV type 1 and 27 with HSV type 2) of the oropharynx and genitalia, caused by the same virus in each person, to evaluate the influence of viral type (HSV-1 vs. HSV-2) and site of infection (oropharyngeal vs. genital) on the frequency of recurrence. The subsequent recurrence patterns of HSV infection differed markedly according to viral type and anatomical site. Oral-labial recurrences developed in 5 of 12 patients with HSV-1 and 1 of 27 patients with HSV-2 (P less than 0.001). Conversely, genital recurrences developed in 24 of 27 patients with HSV-2 and 3 of 12 patients with HSV-1 (P less than 0.01). The mean rate of subsequent genital recurrences (due to HSV-1 and HSV-2) was 0.23 per month, whereas the mean rate of oral-labial recurrences was only 0.04 per month (P less than 0.001). The mean monthly frequencies of recurrence were, in order, genital HSV-2 infections, 0.33 per month; oral-labial HSV-1 infections, 0.12 per month; genital HSV-1 infections, 0.020 per month; and oral HSV-2 infections, 0.001 per month (P less than 0.01 for each comparison). We conclude that the likelihood of reactivation of HSV infection differs between HSV-1 and HSV-2 infections and between the sacral and trigeminal anatomical sites. The sixfold more frequent clinical recurrence rate of genital HSV infections as compared with oral-labial HSV infections may account for the relatively rapid increase in the prevalence of clinically recognized genital herpes in recent years. PMID- 3033507 TI - Do vasodilators prolong life in heart failure? PMID- 3033505 TI - Young children as a probable source of maternal and congenital cytomegalovirus infection. AB - To identify possible sources of cytomegalovirus infection in pregnant women, we studied seven families with a recent case of congenital or maternal cytomegalovirus infection and a history of maternal contact with a young child shedding the virus. We used restriction-endonuclease techniques to compare the DNA of viral isolates collected from family members. Five families contained an infant who had congenital or perinatal infection, a mother who had had evidence of primary infection during her most recent pregnancy, and a child less than three years of age who was excreting cytomegalovirus. All five of the young children attended day-care centers at least part-time. In each of these five families, strains from family members were identical, and it is most likely that the toddler-aged child was the source of the virus for both the mother and the fetus or infant. In two other families, acquisition of cytomegalovirus by children in a day-care center was followed by seroconversion in the mother along with excretion of a strain of the virus identical to that in her child, as demonstrated by restriction-endonuclease analysis. Five of the seven fathers were tested for antibody to cytomegalovirus; four were seronegative, ruling them out as a source of infection in the mothers. These results not only strengthen evidence for the transmission of cytomegalovirus from child to mother but also indicate that infections acquired by a mother from a child can be transmitted to her fetus. PMID- 3033508 TI - Striking the balance on AIDS. PMID- 3033509 TI - Regulation of transcription. Are some controlling factors more equal than others? PMID- 3033511 TI - Role of arginine-tRNA in protein degradation by the ubiquitin pathway. AB - Degradation of intracellular proteins through the ubiquitin and ATP-dependent proteolysis pathway involves several steps. Initially, ubiquitin is covalently linked to the proteolytic substrate in an ATP-requiring reaction. Proteins marked by ubiquitin may then be selectively lysed in a reaction that also requires ATP (for reviews see refs 1-3). A major question concerns the structural features of a protein that make it a specific substrate for ubiquitin-mediated degradation. It was shown that a free alpha-NH2 group is one important feature of the protein structure recognized by the ubiquitin ligation system, and that the half-life in vivo of a protein with an exposed amino terminus depends on its amino terminal residue. We have previously demonstrated that transfer RNA (tRNA) is essential for conjugation of ubiquitin and for the subsequent degradation of proteins with acidic amino termini (aspartate or glutamate). We now show that tRNA is required for post-translational conjugation of arginine to acidic amino termini of proteins, a modification that is essential for their degradation by the ubiquitin pathway. PMID- 3033510 TI - Two growth factor signalling pathways in fibroblasts distinguished by pertussis toxin. AB - The primary action of a family of mitogens including bombesin, bradykinin, vasopressin and alpha-thrombin is to activate the hydrolysis of polyphosphoinositides. Hydrolysis of phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2) by phospholipase C is mediated through coupling of surface receptors to a GTP-binding protein (Gp protein) which, in some cells, is inactivated by the toxin of Bordetella pertussis. It is not known whether this signalling pathway is involved in initiating DNA replication, whereas it has been firmly established that reinitiation of DNA synthesis can be triggered without activation of PtdIns(4,5)P2 hydrolysis by, for example, EGF (epidermal growth factor), FGF (fibroblast growth factor) and insulin/IGF-I (insulin-like growth factor-I), members of a class of mitogens known to activate receptor tyrosine kinases. Taking advantage of the fact that Chinese hamster lung fibroblasts respond to either class of mitogens and that their Gp protein appears to be sensitive to pertussis toxin, we have now analysed the toxin's effect on reinitiation of DNA synthesis and find that it inhibits up to 95% of thrombin induced mitogenicity without affecting EGF- or FGF-induced DNA synthesis and proliferation. These findings strongly suggest that activation of PtdIns(4,5)P2 phospholipase C has a determinant function in growth control, and confirm the existence of alternative growth factor-signalling pathways independent of polyphosphoinositide breakdown. PMID- 3033513 TI - Cross-talk between cellular signalling pathways suggested by phorbol-ester induced adenylate cyclase phosphorylation. AB - Receptor-mediated activation of both adenylate cyclase and phosphatidylinositide hydrolysis systems occurs through guanine nucleotide regulatory proteins and ultimately leads to specific activation of either cyclic AMP-dependent protein kinase A or Ca2+/phospholipid-dependent protein kinase C. Given the remarkable diversity of agents that influence cellular metabolism through these pathways and the similarities of their components, interactions between the two signalling systems could occur. In fact, stimulation of cells with 12-O-tetradecanoyl phorbol-13-acetate (TPA), a phorbol ester that activates protein kinase C, influences hormone-sensitive adenylate cyclase. In some cells TPA induces desensitization of receptor-mediated stimulation of adenylate cyclase, whereas in others, such as frog erythrocytes, phorbol ester treatment results in increased agonist-stimulated as well as basal, guanine nucleotide- and fluoride ion stimulated adenylate cyclase activities. We show here that TPA produces phosphorylation of the catalytic unit of adenylate cyclase in frog erythrocytes. Moreover, purified protein kinase C can directly phosphorylate in vitro the catalytic unit of adenylate cyclase purified from bovine brain. These results suggest that phosphorylation of the catalytic unit of adenylate cyclase by protein kinase C may be involved in the phorbol ester-induced enhancement of adenylate cyclase activity. In addition to providing the first direct demonstration of a covalent modification of the catalytic unit of adenylate cyclase, these results provide a potential biochemical mechanism for a regulatory link between the two major transmembrane signalling systems. PMID- 3033514 TI - Histamine receptors branch out. PMID- 3033512 TI - Tumour prevention and rejection with recombinant vaccinia. AB - Tumour-specific antigens (TSA; ref. 1) have been exploited in the diagnosis and imaging of human cancer and anti-TSA antibodies have therapeutic potential. Vaccination with TSA or anti-idiotypic (TSA) antibodies has also been used to control tumour growth in model systems. An effective immune response nevertheless demands copresentation of antigen with host histocompatibility determinants. We therefore examined whether live vaccinia virus recombinants expressing TSA in cells of the vaccinated host might better elicit tumour immunity. Polyoma virus (PY) is tumorigenic in rodents; because killed PY-transformed cells can elicit tumour immunity, a PY-specific TSA has been postulated. Tumorigenesis involves expression of three early PY proteins, large-T (LT), middle-T (MT) and small-T (ST), but their role as TSAs is unclear. We therefore expressed the three T proteins in separate vaccinia recombinants. Rejection of PY tumours was observed in rats immunized with recombinants expressing either LT or MT. Further, tumour bearing animals could be induced to reject their tumours by inoculation of recombinants. PMID- 3033515 TI - Origins of HTLV-4. PMID- 3033517 TI - A novel class (H3) of histamine receptors on perivascular nerve terminals. AB - Two types of histamine receptor, the H1- and H2-receptors, are found not only on vascular smooth muscle cells but on the perivascular autonomic nerve terminals. Activation of the prejunctional histamine receptors modifies transmitter release from the nerve terminals. Recently, histamine was shown to inhibit its own release from depolarized slices of rat cerebral cortex. This phenomenon was found to be mediated by a novel class of histamine receptor, the H3-receptor, that was pharmacologically distinct from the H1- and H2-receptors. Up to now, there has been no indication whether this third class of histamine receptor is present in any tissue other than the brain. We report here that histamine depresses sympathetic neurotransmission in the guinea-pig mesenteric artery by interacting with histamine H3-receptors on the perivascular nerve terminals. The pharmacological properties of these receptors are similar to those reported for the H3-receptors in the brain. Our data provide evidence for the existence of H3 receptors in the autonomic nervous system. PMID- 3033518 TI - Call for destruction of smallpox virus. PMID- 3033516 TI - Highly potent and selective ligands for histamine H3-receptors. AB - New drugs selective for histamine H3-receptors can be used to establish that these receptors are involved in the feedback control of histamine synthesis and release, and to demonstrate their distribution in the brain and peripheral tissues. These drugs provide new tools for affecting physiological and possibly pathological conditions in which histamine is involved. PMID- 3033519 TI - Ectoenzymes control adenosine modulation of immunoisolated cholinergic synapses. AB - One of the most important inhibitory modulators of synaptic transmission in mammalian brain is adenosine. At some cholinergic terminals, adenosine is known to inhibit further release of acetylcholine. It is unclear whether adenosine is released directly at the synapse or whether ATP is co-released with transmitter and hydrolysed to adenosine in the synaptic cleft. Methods used in the past for isolating nerve terminals have not yielded homogeneous preparations, making it impossible to determine whether sufficient ATP or adenosine is released at specific synapses for inhibition of transmitter release to occur. Immunoaffinity purification techniques have recently permitted the preparation of homogeneous populations of cholinergic nerve terminals, which release ATP upon stimulation. We now report that in immunoisolated cholinergic nerve terminals from the striatum synaptic ectophosphohydrolases convert this ATP to adenosine, which inhibits further acetylcholine release, but this inhibitory effect is not seen in cortical cholinergic terminals lacking the complete ectophosphohydrolase pathway. Therefore the differing adenosine-mediated modulation in different brain areas is controlled by the presence and activity of synaptic ectophosphohydrolases. PMID- 3033520 TI - Isolation of an excision product of T-cell receptor alpha-chain gene rearrangements. AB - The genes for the T-cell receptor, like the immunoglobulin genes, are rearranged as DNA. The mechanism of this rearrangement is not clear; unequal crossover between chromosomes and the looping-out and excision of the excess DNA have both been suggested. We isolated small polydisperse circular (spc) DNAs from mouse thymocytes and cloned them into a phage vector. Of the 56 clones we analysed, nine contained sequences homologous to T-cell receptor alpha-chain joining (J alpha) segments. We have characterized one of these clones; it contains one J alpha segment, and the product out of the recombination of a variable region of the alpha-chain gene (V alpha) with a J alpha gene segment. This is the first demonstration of the presence in extrachromosomal DNA of a reciprocal recombination product of any rearranging immunoglobulin or T-cell receptor gene. The finding verifies that V alpha-J alpha joining can occur by the looping-out and excision of chromosomal DNA. PMID- 3033521 TI - Are the alpha 2-adrenoceptors in the noradrenaline cell body region of physiological significance? AB - Preganglionic stimulation of the cervical sympathetic increased the content of 3,4-dihydroxyphenylacetic acid (DOPAC) in the superior cervical ganglion of rats treated with the dopamine beta-hydroxylase inhibitor FLA-63. It also increased and decreased the concentrations of dopamine and noradrenaline, respectively, in the salivary gland. The effects were partially inhibited by the alpha 2 adrenoceptor agonist clonidine via a yohimbine-sensitive mechanism. The alpha 2 adrenoceptor antagonist yohimbine by itself did not enhance the stimulation evoked increase in ganglionic DOPAC, but it markedly potentiated the stimulation evoked changes in dopamine and noradrenaline in the salivary gland. The results indicate that there are inhibitory alpha 2-adrenoceptors both in the somatodendritic and in the axon terminal region of the noradrenaline neurons but that only the alpha 2-receptors of the axon terminals are physiologically stimulated. PMID- 3033523 TI - Inotropic and electrophysiological effects of 8-substituted cyclic AMP analogues on guinea-pig papillary muscle. AB - The inotropic potencies of 8-substituted cyclic AMP analogues, applied as sodium salts and in form of benzyl esters, were determined in isolated guinea-pig papillary muscles contracting isometrically at a frequency of 0.2 Hz. Half maximally effective concentrations, EC50, for the positive inotropic effect of 8 substituted cyclic AMP (sodium salt) increased in the order 8-(4-chloro phenyl)thio-cyclic AMP, 8-tertiary-butyl-thio-cyclic AMP, 8-benzyl-seleno-cyclic AMP, 8-benzyl-thio-cyclic AMP, 8-methyl-thio-cyclic AMP, 8-bromo-cyclic AMP. Neutralization of the phosphate hydroxyl residue of 8-substituted cyclic AMP by a benzyl group yielded cyclic AMP benzyl esters (cAMP-O-Bn) which were 30 to 100 times more potent than the respective cyclic AMP salts. Cyclic AMP derivatives with a 8-(4-chloro-phenyl)thio- or a 8-tertiary butyl-thio substituent showed comparatively high inotropic potencies. The intrinsic activity was uniformely the same for all 8-substituted cyclic AMP derivatives and equalled that of isoprenaline. As measured by octanol/water partitioning (log P), the increase in lipophilicity of 8-substituted cyclic AMP by esterification with a benzyl group was 7000-fold for 8-bromo-cyclic AMP, 5000-fold for 8-methyl-thio-cyclic AMP, and approximately 1000-fold for the other derivatives. Within the series of benzyl esters, differences in lipophilicity were small. The positive inotropic effect of 8-substituted cyclic AMP analogues was accompanied by a shortening of contraction duration, mainly due to an abbreviation of relaxation time.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3033522 TI - Age-related alterations in alpha 1- and beta-adrenoceptors mediated responsiveness of rat aorta. AB - We have examined the responsiveness of alpha- and beta-adrenoceptors in aortae from 1.5, 3, 6 and 24 month old rats. The isometric contraction to phenylephrine was antagonised competitively by prazosin with a pA2 value of 9.45, suggesting that the receptor is an alpha 1-adrenoceptor. The potency of phenylephrine was significantly reduced in 24 months old as compared with all younger rats combined. The maximum contraction to phenylephrine was unaltered in 24 month old rats. The maximum contraction to potassium chloride was significantly less than that to phenylephrine only in 1.5 months old rats. In tissues contracted by potassium chloride, isoprenaline produced a marked relaxation in 1.5 months old animals, but there was a progressive loss with increasing age of the beta adrenoceptor-mediated relaxation which was markedly reduced by 6 months and abolished in 24 months old. It is concluded that, in the rat aorta, there is a decrease in alpha 1-adrenoceptor responsiveness in senescence, and a loss of beta adrenoceptor-mediated responses in maturation. PMID- 3033524 TI - [Pharmacologic study of the inhibition of the glomerular input to the olfactory tubercle of the frog induced by a puff of air on the olfactory mucosa]. AB - The effects of 4-aminopyridine (10(-2) mol/l), aminooxyacetic acid (AOAA, 10(-4) 10(-3) mol/l), beta-alanine (10(-3)-10(-2) mol/l) and bicuculline (10(-5), 10(-4) mol/l) applications on the frog olfactory bulb (OB) were studied in vivo. The suppression of the orthodromic potential postsynaptic components evoked by a puff on the olfactory mucosa (OB input inhibition), or by single electrostimulation of the olfactory nerve (postsynaptic inhibition) was evaluated. 4-aminopyridine greatly reduced the OB input inhibition and strongly increased the postsynaptic inhibition. AOAA and bicuculline increased and beta-alanine slightly reduced the OB input inhibition. Clear-cut, simple alterations of the postsynaptic inhibition under the influence of the same drugs were not observed. The results confirm the hypothesis, that the OB input inhibition evoked by a puff on the olfactory mucosa may appear due to the release of GABA from glial cells and its binding with presynaptic GABAB-receptors in glomeruli. PMID- 3033525 TI - [Relation between the surface potential of mouse neuroblastoma clone C1300 cells and the phase of the cell cycle]. AB - The surface charge of neuroblastoma cells in different phases of the cell cycle was studied by the microelectrophoresis method. The surface charge increased by 50% on the average after addition of colchicine to the culture medium, by 20-30% after addition of dibutyryl-cAMP or removal of the serum from the medium and decreased by 30% after addition of DMSO. These changes correlated well with variations of the protein content per cell. PMID- 3033526 TI - [Cumulative effect of alcohol (ethanol) on the activity of the cerebellar Purkinje cells of the cat]. AB - The activity of the identified cerebellar Purkinje cells after three successive intravenous injections of 2 ml/kg of 30% alcohol (ethanol) solution was studied in anaesthetized cats. After the first injection a sharp increase in the discharge frequency of Purkinje cells activated by mossy fibre afferent input was accompanied by a decrease in the discharge frequency of the Purkinje cells activated by climbing fiber input. Each successive injection of the same dose of alcohol produced a cumulative effect on the activity of the cerebellar Purkinje cells. PMID- 3033527 TI - [Growth of neurites and formation of connections in cultures of pteropodial mollusc neurons]. AB - Dissociated neurons from the brain of pteropodial mollusc were cultivated in a 25% Leibovitz medium containing 2% of calf serum. Neurite outgrowth was observed in 1-30% of the neurons. It was maximum during the first 3 days. Neurite length reached 300 microns. Membrane potential of neurons was 40-60 mV; they generated single spikes or bursts of impulses. Intercellular connections were tested on the 3-4th days in 70 pairs of neurons with neurites overlapped. Electrical connections between the cells were observed in 20% of the pairs tested, and in 6% of pairs stimulation of one of the neurons evoked an inhibitory postsynaptic potential in the other. PMID- 3033528 TI - [Augmentation of the atrophic mandibular alveolar process with hydroxylapatite granules. Preliminary results]. PMID- 3033529 TI - Nephropathy associated with Wilms' tumor. A case of a 13-year-old girl. AB - The association of nephropathy with Wilms' tumor and male pseudohermaphroditism is known as Drash's syndrome. The absence of either pseudohermaphroditism or Wilms' tumor has also been reported. The nephropathy, characterized by proteinuria or nephrotic syndrome, leads rapidly and inevitably to renal failure. The renal findings are those of a chronic glomerulopathy with mesangial hypercellularity, glomerular sclerosis, interstitial infiltration and marked tubular dilation. This infrequent syndrome usually appears in early life, between 1 and 3 years of age. The case of nephropathy associated with Wilms' tumor we present here is characterized by age and onset: an abrupt onset of renal insufficiency without previous signs of nephropathy in a 13-year-old girl. PMID- 3033530 TI - Phenobarbital and 6-aminonicotinamide effect on cerebral enzymatic activities related to energy metabolism in different rat brain areas. AB - The effect of phenobarbital (100 mg/kg i.p.) and 6-aminonicotinamide (6AN) (35 mg/kg i.p.) on enzyme activities related to energy transduction was investigated on the homogenate "in toto", non-synaptic mitochondrial fraction and synaptosomal fraction isolated from different rat brain areas (cerebral cortex, hippocampus, hypothalamus, striatum, and medulla oblongata). 6AN treatment decreased: phosphofructokinase in all the areas tested; lactate dehydrogenase on the homogenate "in toto" in striatum and hypothalamus, and on the synaptosomal fraction in cerebral cortex and corpus striatum; succinate dehydrogenase on non synaptic mitochondrial fraction in hippocampus and striatum. Finally, aspartate aminotransferase was increased on non-synaptic mitochondrial fraction in striatum and medulla oblongata. Phenobarbital treatment induced an increase of total NADH cytochrome c reductase on mitochondrial fraction in hippocampus and hypothalamus, and a decrease of cytochrome oxidase activity on non-synaptic mitochondrial fraction in hypothalamus and medulla oblongata. PMID- 3033531 TI - The molecular structure of opiate receptors. AB - Examples are given which demonstrate that the kappa opiate receptor can be separated from mu and delta subtypes by their physical parameters. When the subunit composition of the subtypes are compared, no definite differences are encountered. The data from the literature are also contradictory. This may in part be explained by the fact that the different receptors appear to contain a structurally common high affinity binding site. A possible hypothesis would be that the subtypes differ from each other by the number of subunits. PMID- 3033532 TI - Neurochemical evidence for two types of presynaptic alpha 2-adrenoceptors. AB - Neurochemical and pharmacological evidence has been obtained that noradrenergic varicosities (in mouse and rat vas deferens) and cholinergic varicosities (in the Auerbach's plexus) contain heterogenous alpha 2-adrenoceptors through which the release of [3H]noradrenaline and [3H]acetylcholine can be modulated. The quantitative data also support the hypothesis that different noradrenaline and xylazine sensitive alpha 2-adrenoceptors are present prejunctionally in the vas deferens and Auerbach's plexus preparations. Prazosin, although it has a presynaptic inhibitory effect on alpha 2-adrenoceptors of noradrenergic axon terminals, has no effect on cholinergic axon terminals. These data suggest that there are two different types of alpha 2-adrenoceptors at the presynaptic axon terminals. PMID- 3033533 TI - The hydrolysis of glycerophosphocholine by rat brain microsomes: activation and inhibition. AB - Experiments with glycerophosphocholine phosphodiesterase (GPC diesterase, EC 3.1.4.2.) in rat brain microsomes suggest that, although its activity is inhibited by low concentrations of calmidazolium, its dependence on Ca2+ ions is not modulated by calmodulin. The activity of glycerophosphocholine choline phosphodiesterase (choline phosphohydrolase, EC 3.1.4.38) was much lower than that of the GPC diesterase. A relatively inexpensive method for the preparation of sn-glycero-3-phospho [Me-14C]choline is described. PMID- 3033534 TI - Effects of specific activation of mu-, delta- and kappa-opioid receptors on the secretion of luteinizing hormone and prolactin in the ovariectomized rat. AB - With the recent development of highly specific ligands for the mu, delta and kappa opioid receptors it was of interest to define the effects of activation of each of these receptor types on LH and prolactin (PRL) secretion. The compounds were infused (10 microliters/h) at various concentrations into the third cerebroventricle of unanesthetized, ovariectomized rats. The mu agonist, DAGO, at both 1 and 10 micrograms/h caused a significant suppression of LH secretion and a significant stimulation of PRL release. DPDPE, the delta agonist, had no effect on either hormone at 1 microgram/h but inhibited LH secretion at 10 micrograms/h. There was still no effect of this high dose of DPDPE on PRL release. The kappa agonist, U50,488H, had no effect on either hormone at 10 micrograms/h, but at 100 micrograms/h produced a significant suppression of LH release and a highly variable increase in PRL. Coinfusion of 100 micrograms/h of naloxone with the high dose of each of the agonists completely blocked the responses of both hormones to each of the agonists with one exception: the highly variable stimulation of PRL by U50,488H was not affected, thus indicating a nonspecific effect of U50,488H on PRL secretion. These results demonstrate that: activation of the mu receptors produces an inhibition of LH secretion and a stimulation of PRL release; activation of the delta receptors produces an inhibition of LH secretion but has no effect on PRL release, and activation of the kappa receptors produces an inhibition of LH release and a variable stimulation of PRL secretion. PMID- 3033535 TI - Characterization and modulation of corticotropin-releasing factor in the neurointermediate pituitary gland. AB - The present study attempts to determine whether part of the corticotropin releasing factor (CRF)-like materials present in the 'posterior pituitary' is composed of authentic CRF and examines whether the concentration of that peptide may be modulated by circulating glucocorticoids. Analysis of crude extracts of neurointermediate lobes (NIL) of rat pituitaries by reverse-phase HPLC, coupled with a specific radioimmunoassay (RIA), revealed the presence of a major component eluting with the same retention time as rat CRF (rCRF) and, importantly, which was indistinguishable by RIA from the synthetic peptide. Also, two minor forms eluted earlier than rCRF upon HPLC; one of these forms matched the elution position of r[Met(O)21,38]CRF. All three species did show biological activity and stimulated ACTH release from pituitary cells. Essentially the same elution profile was generated by median eminence (ME) extracts. Immunoreactive CRF (CRFi) content of the NIL was about 3% of that of the ME and was found to undergo a significant increase as a result of long-term adrenalectomy while, in contrast, CRFi content of the ME was decreased. This effect of adrenalectomy was completely antagonized by dexamethasone treatment. This study thus provides strong evidence for the presence of authentic CRF within the NIL of the rat pituitary and also shows that tissue concentration of that peptide was modulated by glucocorticoids. PMID- 3033536 TI - Effects of dexamethasone on central and peripheral ACTH systems in the rat. AB - To investigate the simultaneous effects of dexamethasone on peripheral and central adrenocorticotropic hormone (ACTH) systems, rats were treated with dexamethasone or saline for 4 days. Pituitary, plasma, hypothalamus and cerebrospinal fluid (CSF) were then collected and analyzed for ACTH immunoreactivity. Additionally, hypothalamic tissue extracts were analyzed for corticotropin-releasing hormone (CRH) immunoreactivity. Dexamethasone significantly lowered peripheral levels of ACTH as measured in pituitary and plasma. Hypothalamic ACTH content significantly increased while CSF ACTH significantly decreased with dexamethasone treatment. Hypothalamic CRH concentrations showed a small but statistically insignificant decrease. These results suggest that prolonged exposure to dexamethasone affects central as well as peripheral ACTH activity, corroborate our previous findings in rhesus monkeys of decreased CSF ACTH in response to prolonged dexamethasone treatment, suggest that dexamethasone may inhibit the release of ACTH from hypothalamic neurons into the CSF, and provide evidence that the effect of dexamethasone on pituitary ACTH content is of greater magnitude than its effect on hypothalamic CRH. PMID- 3033537 TI - Pituitary secretions related to adrenocorticotropic hormone induce sensitivity of adipose tissue to the insulin-like actions of growth hormone. AB - In its initial encounter with growth hormone (GH) in vitro, epididymal fat excised from GH-deficient rats responds with an insulin-like increase in glucose metabolism. Tissues freshly excised from normal rats are refractory to the insulin-like effects of GH, but become sensitive immediately after surgical stress. Reversal of refractoriness is prevented by administration of the opioid antagonist, naloxone, just prior to stress, suggesting a possible role of beta endorphin or related peptides. These experiments were undertaken to determine the source of these peptides which might equally well be released from the pituitary, adrenal medullae, or nerve endings in response to stress. Since adrenalectomy, like stress, also results in increased secretion of adrenocorticotropic hormone (ACTH) and related peptides, we studied the effects of GH on glucose oxidation in adipose tissue obtained from adrenalectomized rats and found a significant insulin-like response to GH in tissues studied 4 days after adrenalectomy. This effect was not due to GH deficiency, since plasma concentrations were only slightly reduced by adrenalectomy. Administration of naloxone (250 micrograms/rat), 30 or 60 min before sacrifice, or dexamethasone (100 micrograms/injection), 60 and 120 min before sacrifice, prevented a response to GH without affecting circulating levels of GH. The effects of adrenalectomy could not be reproduced by preincubation of adipose tissue from normal nonstressed rats with ACTH and beta-endorphin, but were duplicated by preincubation of adipose tissue for 15 min in medium in which pituitary glands had previously incubated in the presence of corticotropin-releasing hormone (0.1 microM) and arginine vasopressin (0.2 microM). Addition of naloxone (250 micrograms/ml) blocked this effect.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3033538 TI - Effect of catechol estrogens on prolactin secretion in the rat: lack of correlation with estrogenic activity. AB - Temporal changes in plasma prolactin (PRL) levels caused by the administration of a number of different catechol estrogens to freely-moving conscious male rats were determined. The steroids with a wide range of affinity for the estrogen receptor were given in a single bolus (100 micrograms/kg) into the right atrium via an indwelling cannula. Plasma PRL concentration was monitored by taking blood samples every 2 min. The effect on PRL secretion was found to be more pronounced when the catechol estrogens were administered to rats bearing implants of estradiol and could not be correlated with their Kd values for the estrogen receptor. The synthetic estrogen, diethylstilbestrol, behaved anomalously by producing an elevation in plasma PRL only in estradiol-primed animals or after prolonged infusion. Diethylstilbestrol-treated rats showed no PRL response to 2 hydroxyestrone unlike those bearing Silastic capsules of estradiol. The experiments support the hypothesis that catechol estrogens generally act as short term dopamine agonists, lowering PRL, but that those which possess estrogenic activity also show a more prolonged PRL-elevating effect. Thus, the overall action of this group of steroids will be determined by factors which influence these two parameters. PMID- 3033539 TI - Immunoreactive delta sleep-inducing peptide in pituitary adrenocorticotropin/alpha-melanotropin cells and adrenal medullary cells of the pig. AB - Immunoreactive delta sleep-inducing peptide (DSIP) is known to occur in the central nervous system and in body fluids including cerebrospinal fluid, blood and urine. However, the exact nature of the immunoreactive material demonstrated has been a matter of discussion. In the present study, DSIP-like immunoreactivity was demonstrated in the porcine pituitary and adrenal medulla using radioimmunoassay and immunocytochemistry. In the pituitary, the DSIP-like material was present in a subpopulation of the cells storing adrenocorticotropin and alpha-melanotropin. The DSIP immunoreactive cells in the adrenal medulla were identical with a major population of the noradrenaline-storing cells. High performance liquid chromatography of tissue extracts revealed one major peak of DSIP-like immunoreactivity which did not coelute with synthetic DSIP. The immunoreactive material may represent N-terminally truncated fragments of DSIP. The present results suggest that the anterior and intermediate lobes of the pituitary and the adrenal medulla are potential sources of DSIP-like peptides. PMID- 3033540 TI - Sympathoadrenal activity facilitates beta-endorphin and alpha-MSH secretion but does not potentiate ACTH secretion during immobilization stress. AB - The potential involvement of the sympathoadrenal system in stress-induced secretion of peptides from the intermediate lobe of the pituitary gland and the activation of the pituitary-adrenal axis was studied. Male Wistar rats were subjected to control procedures, to sympathectomy by chronic administration (8 weeks) of guanethidine and/or to medullectomy by adrenal enucleation 9 weeks prior to exposure to forced immobilization stress for various periods of time. In intact or sham-operated rats, immobilization caused a prompt increase of circulating norepinephrine, epinephrine (EPI), corticosterone and of immunoreactive adrenocorticotropic hormone (ACTHi), alpha-melanocyte-stimulating hormone (alpha-MSHi) and beta-endorphin (beta-ENDi). Peak levels of pituitary hormones were found after 10 min of stress exposure, but fell to less than 30% of these levels after 2.5 h of immobilization. Adrenal medullectomy, which abolished the stress-induced release of EPI, reduced the acute increase of plasma alpha MSHi and beta-ENDi, but did not influence the acute increase of plasma ACTHi during immobilization stress. Also in medullectomized plus sympathectomized rats, the initial stress response of circulating ACTHi was not different from that of controls. Adrenal medullectomy with or without additional sympathectomy caused a marked increase in plasma ACTHi concentrations after prolonged stress exposure. We conclude that: catecholamines originating from the adrenalmedulla facilitate the stress-induced secretion of intermediate lobe peptides (alpha-MSHi, beta ENDi); catecholamines from the sympathoadrenomedullary system do not contribute to the acute release of ACTH during immobilization stress; the sympathoadrenomedullary system is involved in the secondary reduction of circulating ACTHi levels seen during prolonged stress. PMID- 3033541 TI - Effect of two synthetic alpha-gliadin peptides on lymphocytes in celiac disease: identification of a novel class of opioid receptors. AB - Two synthetic peptides containing residues 43-47 and 43-49 of alpha-gliadin were tested for inhibition of leukocyte migration in 47 patients with celiac disease. In nineteen patients, all on a normal diet, leukocyte migration was inhibited by the peptides and naloxone blocked this effect. In twenty-eight patients (24 of whom were on strict gluten-free diet) leukocyte migration was not affected by the peptides. Our results suggest that alpha-gliadin-(43-49), Tyr-Pro-Gln-Pro-Gln-Pro Phe, is closely related to the active fragment, or to one of the active fragments of alpha-gliadin, and that it interacts with receptors that are similar to but not identical with the known opiate receptors. PMID- 3033542 TI - Distribution and possible origin of beta-endorphin and ACTH in discrete brainstem nuclei of rats. AB - Immunoreactive ACTH and beta-endorphin in the lower brainstem nuclei of intact and brainstem-hemisected rats were quantified by radioimmunoassay. The distribution of these peptides was almost identical throughout the lower brainstem. The concentrations of ACTH and beta-endorphin were essentially equal when expressed on a molar basis. Both peptides were distributed unevenly in the lower brainstem. High concentrations were found in the periaqueductal central gray matter and the dorsal raphe nucleus, and moderate levels were present in the locus coeruleus, the parabrachial nuclei, the nucleus raphe magnus and in the nucleus of the solitary tract. beta-Endorphin was measurable in all 43 brainstem nuclei investigated; ACTH was non-detectable in the red and lateral cuneate nuclei. Except in certain areas in the medulla oblongata (nucleus of the solitary tract, lateral reticular nucleus, reticular formation) ACTH and beta-endorphin declined in brainstem nuclei 10 days after midbrain hemisections. Retrograde accumulation of ACTH and beta-endorphin was found in the arcuate nucleus 3 days after midbrain hemisection, which was mainly ipsilateral to the lesion. Data from brainstem-hemisected rats also indicated that ACTH and beta-endorphin in the nucleus of the solitary tract are primarily of local origin, whereas the lateral reticular nucleus (A1 and A5 catecholaminergic cell groups) and medullary reticular formation may receive ACTH and beta-endorphin innervation from both hypothalamic and medullary neurons. PMID- 3033543 TI - Characterization of dermorphin binding to membranes of rat brain and heart. AB - Binding of dermorphin to the two major opioid receptor types, mu and delta, in rat brain membranes was examined by displacement of [3H] [D-Ala2, MePhe4, Gly (ol)5]enkephalin (DAGO) and [3H]-[D-Ala2,D-Leu5]-enkephalin (DADLE) binding. Affinity of dermorphin binding to mu sites, Kd = 1.24 nM, was almost 3 times greater than that of DAGO, Kd = 3.35 nM. In contrast, the Kd value of dermorphin binding to delta sites was 78 nM only, as compared to Kd = 2.27 nM for DADLE. Dermorphin was ineffective in displacing [3H]ethylketocyclazocine (EKC) binding to kappa receptors after prior blocking of [3H]EKC binding to mu and delta sites. Studies of dermorphin binding to mu sites revealed that the potency of dermorphin increased in the presence of Na+ (+31%) but decreased in the presence of Mn2+ ( 81%) or Gpp(NH)p (-44%). Displacement of bound [3H]diprenorphine (DPN) by dermorphin from atrial membranes of the rat heart, left side, was detectable, suggesting the presence of mu sites in this section of the heart. PMID- 3033544 TI - Neuropathologic and clinical features of Parkinson's disease in Alzheimer's disease patients. AB - While dementia in Parkinson's disease (PD) is well described, PD features in Alzheimer's disease (AD) are being increasingly recognized. In 20 neuropathologically confirmed AD brains, 11 cases (55%) showed PD changes (Lewy body formation, neuronal loss, and gliosis of pigmented nuclei), with no significant difference in age or symptom duration between those cases with and without PD pathology. A history of rigidity in the absence of neuroleptic medication was noted in 80% of those with PD pathology but only 14% of those without PD pathology. Tremor was not observed in either group. This suggests that extrapyramidal signs, especially rigidity, noted in many AD patients are related to coexistent PD pathology. PMID- 3033545 TI - Parkinson's disease and myasthenia gravis: adverse effect of trihexyphenidyl on neuromuscular transmission. AB - A 55-year-old man with idiopathic Parkinson's disease developed myasthenia gravis shortly after taking trihexyphenidyl. The myasthenic weakness waxed and waned with rise and fall in serum levels of trihexyphenidyl, without marked change of anti-acetylcholine receptor antibody titer. PMID- 3033546 TI - Regional nerve injury after intra-arterial chemotherapy. AB - Eleven patients at M.D. Anderson Hospital were referred for neurologic evaluation after having their internal or external iliac arteries catheterized for the treatment of localized pelvic or lower extremity tumors. Nine patients developed lumbosacral plexopathies and two patients, mononeuropathies. All symptoms occurred within 48 hours of the intra-arterial infusion. All patients received cis-dichlorodiammine-platinum (cisplatin; CDDP) intra-arterially, alone or in combination with other agents. Follow-up examinations revealed that only one patient had made partial recovery from the neurologic dysfunction. Chemotherapy induced small vessel injury, with subsequent plexus or nerve infarction, appears to be the most likely cause, although a direct neurotoxic effect of CDDP cannot be excluded. PMID- 3033547 TI - Dexamethasone suppression test abnormalities in multiple sclerosis: relation to ACTH therapy. AB - We studied the 1-mg overnight dexamethasone suppression test (DST) in patients with MS. In about 50% of patients, serum cortisol did not fall below 5.0 micrograms/dl. This percentage was similar in patients with major depression, but contrasted to 11% in normal controls. MS nonsuppressors were not more depressed than suppressors; dexamethasone bioavailability may have contributed because nonsuppressors had lower serum dexamethasone levels than suppressors. Suppressors improved in the week following ACTH therapy; nonsuppressors did not. Furthermore, serum dexamethasone values correlated positively with clinical response to ACTH treatment. The DST may be a useful neuroendocrine test of glucocorticoid sensitivity in MS patients. PMID- 3033548 TI - Polyradiculoneuropathy, polyradiculitis, and CMV in AIDS and ARC. PMID- 3033549 TI - [Ranitidine, cimetidine and magnesium silicate in the prevention of aspiration pneumonia]. PMID- 3033550 TI - [Neuron-specific enolase: a new marker for small-cell pulmonary carcinoma]. AB - A marker of neuroendocrine differentiation, neuron-specific enolase (NSE) is assessed in the diagnosis of small cell lung cancer (SCLC). The market was found to be highly sensitive and extremely specific in high risk groups (smokers and chronic bronchitics). PMID- 3033551 TI - [Undifferentiated large-cell carcinoma of the lung]. AB - Among the series of class 111 cancers of the lung treated in 1980-85 the incidence of large cell carcinoma was 4%. Most of these cases presented lymph node invasion. Smoking does not appear to play a decisive role in pathogenesis. Survival at one year after surgery was nil. The diagnosis and treatment of this form do not permit a satisfactory prognosis. PMID- 3033552 TI - Externally applied adenosine-5'-triphosphate causes inositol triphosphate accumulation in cultured chick myotubes. AB - In striated muscle, adenosine-5'-triphosphate (ATP) potentiates the responses to acetylcholine. The underlying biochemical events are unknown. Here we report that ATP, externally applied to chick myotubes, induces a rapid, dose-dependent accumulation of intracellular inositol triphosphate which is correlated with a decrease in phosphatidyl 4,5-bisphosphate. Adenosine-5'-diphosphate, adenosine-5' monophosphate and adenosine are less potent while beta, gamma-imido ATP is equipotent motoneurons and/or skeletal muscle controls the activation of a polyphosphoinositide phosphodiesterase via a cell membrane P2-purinoceptor, thus modulating skeletal muscle responses to transmitter release. PMID- 3033553 TI - Effects of selective mu- and delta-opioid peptides on kindled amygdaloid seizures in rats. AB - Opioid systems seem to be implicated in the regulation of brain excitability, though in an apparently controversial way. In order to assess the involvement of mu- and delta-opioid receptors in the anti-epileptogenic properties of opioids, i.v. administrations of morphine, and of DAGO (Tyr-D-Ala-Gly-N-Me-Phe-Gly-ol) and DTLET (Tyr-D-Thr-Gly-Phe-Leu-Thr), two peptides presenting selective agonist properties towards respectively the mu and the delta receptors, were performed on fully kindled rats. It is concluded that the mu- rather than the delta-receptors are implicated in the limitation of amygdaloid kindled seizures. PMID- 3033555 TI - Neurotransmission in the pedunculopontine nucleus and pilocarpine-induced motor limbic seizures in rats. AB - Systemic injection of the cholinergic agonist, pilocarpine (380 mg/kg, i.p.) initiates a sequence of events leading to motor limbic seizure activity. Focal injection of the excitatory amino acid antagonist, 2-amino-7-phosphonoheptanoic acid (50 pmol-1 nmol) into the pedunculopontine nucleus (PPN), prior to pilocarpine injection, results in a powerful anticonvulsant action. The GABA agonist, muscimol (25-50 pmol) also afforded protection against pilocarpine evoked convulsions when injected focally into the PPN. The results suggest that an overall inhibition of PPN output neurons is required for anticonvulsant action. PMID- 3033554 TI - Neuropeptide Y receptors and the inhibition of adenylate cyclase in the human frontal and temporal cortex. AB - The presence of a single class of high affinity, saturable binding sites for [3H]neuropeptide Y (NPY) was demonstrated in membranes from human frontal and temporal cortex. The specific binding of [3H]NPY was sensitive to guanosine 5' triphosphate (GTP) and guanylyl-imidodiphosphate (GMPP(NH)P; 100 microM) which lowered the total binding capacity (Bmax) value (35 +/- 2 fmol/mg in the frontal cortex and 82 +/- 3 fmol/mg in the temporal cortex) by 50%. GTP and GMPP(NH)P did not affect the dissociation constant (Kd) value which was 0.25 +/- 0.03 nM in a frontal cortex sample and 0.76 +/- 0.06 nM in the sample from the temporal cortex. The affinity and GTP sensitivity of the [3H]NPY binding to human brain membranes parallels that found in the rat brain. It was demonstrated that occupancy of NPY receptors by NPY (1 microM) inhibits the basal and forskolin (10 microM)-stimulated adenylate cyclase activity by 18-30% in a crude membrane preparation from human frontal cortex. PMID- 3033556 TI - Striatal adenylate cyclase-inhibiting dopamine D2 receptors are not affected by the aging process. AB - Radioreceptor binding studies with various labelled ligands and positron emission tomography have revealed a decline in D2 receptor concentration with age in both animal and human caudate nucleus. In this study we found that during senescence the functional characteristics of D2 receptors that inhibit adenylate cyclase (AC) are unchanged, by measuring the extent of inhibition of AC activity by dopamine mimetic drugs as a direct indicator of D2 receptor function. PMID- 3033557 TI - Phorbol esters broaden the action potential in CA1 hippocampal pyramidal cells. AB - Intracellular recordings were made from CA1 pyramidal cells in rat hippocampal slices. Single action potentials were elicited by injection of brief current pulses. Bath application of phorbol esters (4 beta-phorbol-12,13-diacetate, 0.3-5 microM; or 4 beta-phorbol-12,13-dibutyrate, 5-10 microM) broadened the action potential in each of the cells tested (n = 9). The broadening reflected slowing of the repolarization, whereas the upstroke of the spike was unchanged. This effect may enhance transmitter release from synaptic terminals, and contribute to enhancement of synaptic transmission through activation of protein kinase C, a mechanism which has been associated with long term potentiation. PMID- 3033558 TI - Single sensory neurons activate excitatory amino acid receptors in the lamprey spinal cord. AB - The effects of excitatory amino acid (EAA) antagonists were tested on sensory afferent excitation in the lamprey spinal cord. Paired intracellular recordings were made from mechanosensory neurons (dorsal cells) and second order sensory neurons (giant interneurons). Stimulation of individual dorsal cells evoked mono- and/or polysynaptic excitatory postsynaptic potentials (EPSPs) in giant interneurons. These EPSPs were depressed by the EAA antagonists cis-2,3 piperidine dicarboxylate and kynurenic acid. Small components of the synaptic potentials were due to electrical coupling since they were not depressed by EAA antagonists or by calcium-free solution. The N-methyl-D-aspartate antagonist 2 amino-5-phosphonovalerate did not depress short-latency EPSPs. Thus, mechanosensory neurons in the lamprey spinal cord activate EAA receptors (kainate/quisqualate receptors) on interneurons, via mixed chemical and electrical synapses. PMID- 3033559 TI - Changes of cytochrome oxidase activity in the rat subcortical visual centers after unilateral eye enucleation. AB - Histochemical densitometric measurements of cytochrome oxidase (CO) activity in the dorsal and ventral lateral geniculate nuclei (LGNd and LGNv) and the superficial gray stratum of the superior colliculus (SC) were done between 2 and 12 weeks after unilateral enucleation in the adult albino rat. This deafferentation resulted in a transient, slight but statistically significant decrease in CO activity in the contralateral LGNd and LGNv. Whereas in the contralateral SC, a marked reduction in CO activity was observed, and this persisted over the postoperative periods studied. The regional difference in the recovery pattern of CO activity were discussed from the aspect of synaptic reorganization. PMID- 3033560 TI - Autoradiographic distribution of non-dopaminergic binding sites labeled by [3H]haloperidol in rat brain. AB - The regional distribution of binding sites labeled by [3H]haloperidol, in the presence of excess spiroperidol, was compared to the regional distribution of receptors labeled by [3H]SCH 23390 and [3H]sulpiride, [3H]SCH 23390 and [3H]sulpiride labeled distinct nuclei, such as the olfactory tubercle, caudate, globus pallidus, substantia nigra, and inferior and superior colliculi. In contrast, the distribution of binding sites labeled by [3H]haloperidol, in the presence of excess spiroperidol, were much more extensive. Some areas containing the highest density of sites labeled by [3H]haloperidol were the external plexiform layer of the olfactory bulb, the cerebral cortex, the paraventricular nuclei, the interpeduncular nucleus and the superior colliculus. The distribution of non-dopaminergic binding sites labeled by haloperidol was clearly quite different from that labeled by dopaminergic ligands. PMID- 3033561 TI - Distribution of neuropeptide Y receptors in the rat hippocampal region. AB - The distribution of binding sites for neuropeptide Y (NPY) was studied in the rat hippocampal region by using [3H]NPY together with quantitative in vitro receptor autoradiography. The highest density of specifically bound [3H]NPY was found in regio superior and regio inferior of Ammon's horn. Within these fields, stratum oriens, stratum pyramidale and stratum radiatum harboured the highest densities of [3H]NPY binding while stratum moleculare was relatively poor in [3H]NPY binding sites. In area dentata, the highest density of [3H]NPY binding was found in the inner one third of the molecular layer. In the presubiculum and in the entorhinal area, the outer two layers were slightly more enriched in [3H]NPY binding sites than were the deep layers. In all hippocampal subfields a clear gradient of increased [3H]NPY binding was found at successively more ventral levels. PMID- 3033562 TI - Localization of glycine uptake and receptors in the cat retina. AB - Immunocytochemistry using a monoclonal antibody against glycine receptors revealed that these receptors in cat retina are confined to the inner plexiform layer (IPL). The outer half of that layer showed strong patchy labelling with some indication of two bands corresponding to the on- and off-sublamina. High affinity uptake of [3H]glycine followed by autoradiography labelled amacrine cells, bipolar cells and the outer portion of the IPL. Silver grains in the IPL were patchily distributed. These results indicate in the cat retina a close match between presynaptic glycinergic elements labelled by high-affinity uptake and the postsynaptic receptor sites for glycine revealed by immunocytochemistry. PMID- 3033563 TI - High Na+ affinity of the Na+,K+ pump in isolated rabbit retinal Muller (glial) cells. AB - Rabbit retinal Muller (glial) cells were isolated by means of papain and mechanical dissociation. In a special perfusion chamber, the cells were penetrated with a recording microelectrode. Membrane potential changes were recorded in response to extracellular application of both high-K+ solutions and of ouabain, and that during perfusion with normal and Na+-free solutions, respectively. In other Muller cell preparations, Na+,K+-adenosine triphosphatase (ATPase) activity was measured using a radiochemical method, and its Na+ dependence was determined. All results strongly suggest that the Muller cell's Na+,K+ pump can be activated in the presence of extremely low amounts of Na+. This provides additional evidence for significant differences between the glial and the neuronal enzyme. PMID- 3033564 TI - Suppressing effect of pertussis toxin on clonidine-induced inhibition of noradrenaline release from cerebral cortical slices of rats. AB - Clonidine at 1.0 microM significantly decreased 20 mM K+-evoked release of L [3H]noradrenaline (NA) from rat cerebral cortical slices preloaded with L-[3H]NA. Inhibitory effects of clonidine, however, were not observed in slices pretreated with 20 micrograms/ml pertussis toxin, an islet-activating protein, together with NAD and adenosine triphosphate. It is suggested that the inhibitory guanine nucleotide-binding protein (Ni) could be involved in alpha 2-adrenoceptor mediated inhibition of NA release from nerve terminals in the central nervous system. PMID- 3033565 TI - The cellular forms and functions of the inositol phospholipids and their metabolic derivatives. PMID- 3033566 TI - Changes in body temperature after administration of antipyretics, LSD, delta 9 THC and related agents: II. AB - Antipyretics, in particular acetaminophen, aspirin and ibuprofen, constitute the single most important class of drugs used therapeutically for an effect on body temperature. Hallucinogens exert prominent actions on the central nervous system, and it is not surprising that, like so many other centrally-acting agents, they too often affect temperature. This compilation primarily covers the considerable amount of data published from 1981 through 1985 on the interactions of these drugs and thermoregulation, but data from many earlier papers not included in a previous compilation are also tabulated. The effects of agents not classically considered as antipyretics on temperatures of febrile subjects are also covered. The information listed includes the species used, the route of administration and dose of drug, the environmental temperature at which experiments were performed, the number of tests, the direction and magnitude of change in body temperature and remarks on special conditions, such as age or brain lesions. Also indicated is the influence of other drugs, such as antagonists, on the response to the primary agent. PMID- 3033567 TI - Breast carcinoma in a man following local trauma. PMID- 3033568 TI - Anomalous DMSA uptake in unilateral renal artery occlusion. PMID- 3033569 TI - Technegas--a new ventilation agent for lung scanning. AB - A simple process using technetium-99m generator eluate in a graphite crucible at 2500 degrees C, produces a structured ultra-fine dispersion of labelled carbon. Particle sizes are 5.0 nm (0.005 micron) and less and adhere to the walls of the alveoli on inhalation. Penetration characteristics are gas-like and the radioactivity per litre of carrier argon allows single breath inhalations of a diagnostic dose. Over 190 patients have been studied including 50 within a formal clinical trial with xenon-133. Results of tomography, dynamic inhalation, and image subtraction using this new agent-'Technegas'--are presented. PMID- 3033571 TI - 99Tcm-DMSA uptake in obstructed kidneys. How inaccurate are the 5 h measurements? AB - Partial obstruction of the left ureter was created in 19 rats and a relative 99Tcm-DMSA uptake was obtained 5 and 24 h after intravenous injection of the tracer. A systematic and variable overestimation of the left to right uptake ratio was found at 5 h, with a mean error of 15.8% (S.D. = 12.2). PMID- 3033570 TI - Incidence, severity and clinical course of right ventricular involvement after acute inferior myocardial infarction; assessment by sequential 99Tcm pyrophosphate scan and gated blood pool scan. AB - To evaluate the incidence, severity and clinical course of right ventricular (RV) involvement after acute inferior myocardial infarction (IMI), 78 patients (pts) with IMI were investigated by both 99Tcm-pyrophosphate (PYP) scan and gated blood pool scan (GBPS). GBPS was performed at admission and 10 days, whereas 99Tcm-PYP scan was performed at 3 to 6 days. RV uptake of PYP was demonstrated in 25 (32%) pts on 99Tcm-PYP scan and RV akinesis or moderate hypokinesis by GBPS was observed in 39 (50%) pts on the acute scan; 25 pts (Group A) with positive RV uptake and 14 pts (Group B) with no RV uptake. In the remaining 39 pts (Group C) had normal RV wall motion. Severely depressed RVEF improved nearly 10 points on the tenth day in Group A (from 30.8 +/- 12.3 to 40.9 +/- 6.7%, p less than 0.01) and Group B (from 35.6 +/- 8.2 to 44.5 +/- 10.5%, p less than 0.01), respectively. Group C showed normal RVEF (from 47.4 +/- 7.6 to 50.1 +/- 10.2%). Fourteen pts of 39 (Groups A and B) who had developed shock or hypotension improved strikingly after appropriate therapy except for one death during their hospital course. Our data demonstrated: some patients with RV dysfunction in IMI do not have severe necrosis as judged by PYP scanning, those with positive RV uptake and depressed RV function show a lower degree of recovery than those with no RV uptake, but start from a lower initial value of RV function, and the combination of 99Tcm-PYP scan and GBPS offers prognostic information in IMI with RV dysfunction. PMID- 3033572 TI - Multicentric human papillomavirus infections of the female genital tract: correlation of viral types with abnormal mitotic figures, colposcopic presentation, and location. AB - Human papillomavirus deoxyribonucleic acid (DNA) was identified by Southern blot hybridization in 21 of 24 patients with multicentric anogenital lesions and in 46 of 61 individual lesions. Type 6/11 was present in nine patients, type 16 in one, an undetermined type in one, and more than one type in ten patients. Mixed types were present in eight of 46 virus-positive individual lesions. Abnormal mitotic figures were found in 16, 87, and 75% of lesions associated with type 6/11, type 16, and mixed types, respectively. Colposcopic presentation or location of lesions was not predictive of viral types. The relatively high rate of mixed human papillomavirus types in multicentric lesions and in single lesions, and the lack of absolute correlation between viral types and abnormal mitotic figures, suggest that lesions should be removed to prevent viral transmission and possible progression to carcinoma. PMID- 3033573 TI - A new staining method using a hematoxylin, alcian blue and periodic acid-Schiff reaction for demonstrating mucins in gastric glands. PMID- 3033574 TI - Retinitis pigmentosa with segmental massive retinal gliosis. An immunohistochemical, biochemical, and ultrastructural study. AB - A morphologic, immunohistologic, and biochemical study was made on the eyes of a 79-year-old woman with clinically documented retinitis pigmentosa (RP). The methods included light and electron microscopy, immunohistologic staining, and biochemical analysis of interphotoreceptor retinoid-binding protein (IRBP) and cyclic nucleotides. Results from a histopathologic examination showed marked equatorial pigmentary retinal degeneration as well as peripheral chorioretinal atrophy corresponding to areas of paving stone chorioretinal changes. An unusual finding was a localized equatorial nodule in the right eye that stained with anti glial fibrillary acidic protein (GFAP) antibodies, and showed lipid infiltrates in its margin and base. The equatorial retina showed marked gliosis of the outer layers. Photoreceptor cells were present only in the posterior retina, macula, and focally, in the far periphery. These areas corresponded to detectable IRBP assessed by immunohistochemical staining and biochemical analysis using the enzyme-linked immunosorbent assay (ELISA). Cyclic nucleotides were reduced in the peripheral retina, in areas of photoreceptor cell loss. PMID- 3033575 TI - Differential diagnosis and surgical management of parapharyngeal masses: review and an unusual illustrative case. AB - Diagnosis and management of swellings of the parapharyngeal space are difficult because of the inaccessibility of the region, which contains part of the parotid gland with major vascular and neural structures that may be subject to disease. An unusual case of a spherical mass with a calcified margin within the parapharyngeal space is presented; it illustrates an approach to diagnosis and management. PMID- 3033576 TI - [Diagnostic possibilities of scintigraphy in various diseases and injuries of the hand]. PMID- 3033577 TI - [Polymorphism of human serum paraoxonase activity in childhood]. PMID- 3033578 TI - [Diagnosis and surgical treatment of nasopharyngeal angiofibroma in the light of our clinical cases]. PMID- 3033579 TI - Pain-inducing laryngeal paraganglioma: report of the ninth case and review of the literature. AB - There are two distinct forms of laryngeal paraganglioma. The patient with the rare Type II has exquisite throat pain that is often clinically confusing. The small size or submucosal location of the lesion often results in a delay in correct diagnosis and treatment. The lesion is very aggressive, even if tiny, generally with widespread metastasis. This tumor should be considered in the differential diagnosis of severe persistent throat pain, especially in the absence of abnormalities on physical or radiologic examination. PMID- 3033580 TI - "Composite" tumor--mixed squamous cell and small-cell anaplastic carcinoma of the larynx. AB - Mixed squamous and oat cell carcinomas (composite neoplasms) of the larynx behave aggressively with a propensity toward early dissemination. Diagnosis may not be made until definitive surgery is performed. The extent of surgery must be determined by the size and site of the primary neoplasm and the physiologic status of the patient. In our experience, surgery has adequately controlled the primary disease. A metastatic work-up is essential. Entry into a comprehensive program employing adjunctive radiotherapy and chemotherapy are strongly recommended. Theories concerning histogenesis remain controversial and include two separate colliding tumors, differentiation from a single cell line, or divergent differentiation of a single cell line. PMID- 3033582 TI - The frequency of distribution according to histological types of lung cancer in the tracheobronchial tree among Turkish patients. PMID- 3033581 TI - The multiple primary lung carcinoma. PMID- 3033583 TI - Combined chemo-surgical treatment of oat cell carcinoma. PMID- 3033584 TI - Clinical and functional development of sarcoidosis considering the results of bronchoalveolar lavage. PMID- 3033585 TI - Morphogenesis of salivary gland tumors. A prerequisite to improving classification. PMID- 3033586 TI - Percutaneous fine-needle aspiration biopsy cytology of the kidney and adrenal. PMID- 3033588 TI - Unusual variants of uterine cervical carcinoma. PMID- 3033587 TI - Cutaneous adnexal tumors and cysts: a review. Part II--Tumors with apocrine and eccrine glandular differentiation and miscellaneous cutaneous cysts. PMID- 3033589 TI - Carcinomas of sweat glands. PMID- 3033591 TI - Morphologic correlates of renal growth arrest in neonatal partial ureteral obstruction. AB - To investigate the morphologic correlates of decreased renal blood flow and growth arrest resulting from chronic partial ureteral obstruction in the neonate, guinea pigs were subjected to unilateral ureteral constriction within the first 2 days of life and were studied at 3 and 8 wk of age. Severity of histologic changes in the obstructed and intact contralateral kidney was assessed by light microscopy. Morphometric glomerular measurements were made using a computerized tracing device. Since contralateral nephrectomy or administration of angiotensin converting enzyme inhibitor (enalapril) result in increased blood flow to the obstructed kidney, morphologic changes were also examined in separate groups of animals subjected to these maneuvers, and were compared to appropriate controls. Ipsilateral chronic partial ureteral obstruction resulted in decreased glomerular volume (p less than 0.0001) and increased glomerular crowding associated with tubular dilatation and progressive glomerular sclerosis, tubular atrophy, and interstitial fibrosis. The intact contralateral kidney underwent hypertrophy with increase in glomerular volume (p less than 0.0001) and decreased glomerular density. Contralateral nephrectomy prevented the decrease in glomerular volume in the obstructed kidney and resulted in decreased glomerular density and reduced tubular atrophy at 3 wk of age. Enalapril prevented the decrease in glomerular volume at 3 wk of age, but glomerular and tubular changes progressed and were unaffected by enalapril at 8 wk. Left kidney glomerular volume was directly related to renal blood flow (r = 0.71, p less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3033592 TI - Human papillomaviruses: associations between laryngeal papillomas and genital warts. PMID- 3033590 TI - Epstein-Barr virus-transformed B-cell line (DV-1) derived from bone marrow of a patient with severe combined immunodeficiency and immunoblastic lymphoma. AB - An Epstein-Barr virus-transformed B-cell line derived from a patient with severe combined immunodeficiency who died of a lymphoreticular malignancy has been characterized. The line derived from bone marrow cultures and designated DV-1 shows surface and cytoplasmic IgM and staining with fluorescent monoclonal antibodies against immunoglobulin heavy and light chains mu, delta, and kappa, Dr, and several other B-cell surface antigens. DV-1 secretes IgM kappa and demonstrates monoclonality on analysis of immunoglobulin heavy and light chain gene rearrangement patterns. Incubation with either phytohemagglutinin or pokeweed mitogen failed to cause significant stimulation of proliferation of DV-1 and another EBV-transformed B-cell line derived from an immunologically normal host (LA-350), whereas incubation with Staphylococcus aureus Cowan strain 1 led to significant inhibition of DV-1 and LA-350. Rabbit IgG antibody specific for human immunoglobulin mu-chains produced a dose dependent stimulation of both lines. The responses of DV-1 and LA-350 to mitogens and anti-mu were not as high as those of normal peripheral blood lymphocytes. This spontaneously derived Epstein-Barr virus-transformed B-cell line from a patient with severe combined immunodeficiency demonstrated functional characteristics similar to a B-cell line derived from an immune competent host. While these cells spontaneously incorporate 200 times more thymidine than normal resting peripheral blood lymphocytes, they retain their ability to be modulated by antiimmunoglobulin, and staphylococcal Cowan strain 1, but are unresponsive to the effects of B-cell growth factor. PMID- 3033593 TI - Immunogenicity and reactogenicity of lowered doses of rhesus rotavirus vaccine strain MMU 18006 in young children. AB - Rhesus rotavirus oral vaccine strain MMU 18006 at a dose of 10(5) plaque-forming units (PFU), a 1:10 dilution of the original undiluted vaccine, is highly immunogenic in young children. Fevers have occurred, however, on Days 3 and 4 following vaccination. This study was conducted to determine whether febrile reactions could be eliminated and immunogenicity maintained by (1) giving smaller doses of vaccine or (2) vaccinating younger infants. Thirty-one children between 3 and 11 months of age received, in a randomized, double blind manner, either 10(4) PFU of vaccine virus, 10(3) PFU of vaccine virus or placebo. All recipients of the 10(4) PFU dose had a seroresponse; however, some degree of immunogenicity was lost with the smaller dose (10(3) PFU). Fevers were observed in recipients of both of the lowered doses of vaccine but the febrile reactions were related to the age of the vaccinee. No infant younger than 5 months of age experienced a temperature elevation, whereas the majority of children older than 5 months had fevers. Our data suggest that the lack of reaction in the younger infants correlates with the presence of prevaccination neutralizing antibody, presumably transplacentally acquired. We conclude that the rhesus rotavirus oral vaccine at a dose of 10(4) PFU is immunogenic and appears to be safe in young infants. PMID- 3033594 TI - Cytomegalovirus and the brain. PMID- 3033596 TI - [A case of androblastoma in a 5-year-old boy]. PMID- 3033595 TI - [Serum ferritin as a possible tumor marker in malignant tumors of children]. PMID- 3033597 TI - [Epstein-Barr virus cerebellitis in an 11-year-old girl]. AB - The authors report an eleven year old girl who manifested predominantly an acute cerebellar syndrome secondary to infection by Epstein Barr Virus. This complication is unusual and males are predominantly affected. The diagnostic and common physiopathological hypothesis are discussed. PMID- 3033598 TI - [Acute parvovirus B 19-induced erythroblastopenia and hereditary spherocytosis. Apropos of 1 pediatric case and review of the literature]. AB - Authors report a new case of acute erythroblastopenia linked to a parvovirus B 19 infection by a 10 years old boy suffering from an hereditary spherocytosis. Aurillac antigen or parvovirus B 19 is one of the smallest virus to be known. It has been well demonstrated in vitro that the virus inhibits especially erythropoiesis but mechanism remains unclear. Systematic vaccination of all children at risk with congenital or acquired chronic haemolytic anaemia should be in the near future the best prophylaxis of parvovirus B 19 infections. PMID- 3033599 TI - Decrease in force potentiation and appearance of alpha-adrenergic mediated contracture in aging rat skeletal muscle. AB - The effect of increasing age on contractile performance and catecholamine receptor activity was investigated in a distal, predominantly fast twitch oxidative glycolytic (FOG) muscle from the plantar surface of the rat hindfoot. The ability of the flexor digitorum brevis (FDB), isolated from anesthetized rats and maintained in vitro, to undergo post-tetanic potentiation and a staircase response declined with age. Potentiation following repetitive stimulation was reduced by 50% in 2 year old rats and eliminated in 3 year old animals. The rate of muscle fatigue during intermittent tetanic stimulation also increased in aging muscles. FDB, regardless of age, did not develop a positive inotropic response to 10(-6) M epinephrine applied in vitro, but 3 year old FDB generated a prolonged contracture. Contracture tension was approximately 25% of twitch tension and was maintained for 2-10 min in the continued presence of catecholamine. Contractures were eliminated by pretreatment with alpha-adrenergic antagonists or by removing Ca2+ from the bathing medium. In addition to decreased contractile capacity, aging muscles acquire a population of alpha-adrenergic receptors which may underlie some of the metabolic and structural changes associated with increasing age. PMID- 3033600 TI - Cross index for improving cloning selectivity by partially filling in 5' extensions of DNA produced by type II restriction endonucleases. AB - A cross index is presented for using the improved selectivity offered by the Hung and Wensink (Nucl. Acids Res. 12, 1863-1874, 1984) method of partially filling in 5'-extensions produced by type II restriction endonucleases. After this treatment, DNA fragments which normally cannot be ligated to one another, can be joined providing that complementary cohesive ends have been generated. The uses of this technique, which include the prevention of DNA fragments (both vector and insert) auto-annealing, are discussed. PMID- 3033601 TI - All foot and mouth disease virus serotypes initiate protein synthesis at two separate AUGs. AB - Translation of the foot and mouth disease virus genome in vitro and in vivo indicated that all seven serotypes initiate protein synthesis at two separate AUGs. Sequence analysis of the region surrounding these AUGs has shown that the efficiency with which the initiating AUG is recognized is dependent on the flanking nucleotides. However, in vitro, the major factor determining which AUG is used is the concentration of Mg2+. PMID- 3033602 TI - Comparison of the conformation of an oligonucleotide containing a central G-T base pair with the non-mismatch sequence by proton NMR. AB - We have recorded NOESY spectra of two non-selfcomplementary undecanucleotide duplexes. From the observed NOEs we do not detect any significant distortion of the helix when a G-C pair is replaced by a G-T pair and the normal interresidue connectivities can be followed through the mismatch site. We conclude that the 2D spectra of the non-exchangeable protons do not allow differentiation between a wobble or rare tautomer form for the mismatch. NOE measurements in H2O, however, clearly show that the mismatch adopts a wobble structure and give information on the hydration in the minor groove for the G-T base pair which is embedded between two A-T base pairs in the sequence. PMID- 3033603 TI - Isolation of a human genomic fragment, co-amplified with c-Ki-ras, that affects plasmid supercoiling in E. coli. AB - Amplification of cellular proto-oncogenes has been implicated in the development of human malignancies. A library was constructed from genomic DNA extracted from a lung tumour, previously shown to carry an amplified c-Ki-ras 2 gene. Using a v Ki-ras probe, a fragment with ras homology was isolated and shown to be amplified in the original tumour DNA to the same level as c-Ki-ras. Studies with human hamster hybrids demonstrated that it is normally located on human chromosome 12 (as is c-Ki-ras). The restriction map of the fragment is different from that of the known Ha, Ki or N-ras genes and its sequence shows evolutionary conservation, as demonstrated by hybridisation to the genomic DNA of several mammalian species. A pUC19 subclone (pK42), carrying a 1.3kb insert, shows supercoil heterogeneity in plasmid preparations, as does a second compatible plasmid introduced into the same bacterial host with pK42. It appears therefore that the subclone is encoding a product that affects DNA topoisomerase activity in E. coli. PMID- 3033605 TI - The untranslated leader of nuclear COX4 gene of Saccharomyces cerevisiae contains an intron. AB - The nuclear gene for subunit IV of cytochrome oxidase (COX4) in Saccharomyces cerevisiae contains a 342 bp intron which is contained entirely within the 5' leader of the message. Splicing of the intron results in removal of several small open reading frames; subsequently, the COX4 AUG becomes the 5' proximal initiation codon. A strain with an rna2- mutation fails to splice mRNA efficiently at restrictive temperature and was used to map the intron splice junctions by RNase protection. Two major mRNA initiation sites were mapped by primer extension of synthetic oligodeoxynucleotides. The splice junctions and internal TACTAAC box conform to consensus sequences previously determined from other yeast introns. One gene for subunit V of cytochrome oxidase (COX5b) has also been shown to contain an intron. The significance of introns in two nuclear genes encoding subunits of cytochrome oxidase is discussed. PMID- 3033604 TI - Effects of VM26 (teniposide), a specific inhibitor of type II DNA topoisomerase, on SV40 DNA replication in vivo. AB - Simian Virus 40 (SV40) infected cells were pulse labeled with (3H) thymidine and chased either in the absence or in the presence of the cytotoxic drug VM26 (teniposide). We investigated the structure of labeled SV40 DNA and found that VM26 had no significant effect on replicative chain elongation but strongly inhibited the conversion of late replication intermediates to mature DNA daughter molecules. The late replicative SV40 DNA intermediates which accumulate in VM26 treated cells contained essentially full length labeled DNA strands. These newly synthesized strands were not part of two catenated interlocked SV40 monomers suggesting that the block occurred prior to the final ligation reaction. Since VM26 is known to be a specific inhibitor of DNA topoisomerase II we conclude that this enzyme is dispensable for the chain elongation of replicating SV40 DNA, but that it is essential for the termination of SV40 DNA replication cycles. PMID- 3033606 TI - The action of a water-soluble carbodiimide on adenosine-5'-polyphosphates. AB - When aqueous solutions of adenosine-5'-mono-, di-, or triphosphates are treated with a water soluble carbodiimide the major product is the expected diadenosine 5'-5'-polyphosphate. The yields of these pyrophosphates are greatly increased in the presence of the Mg2+ ion. Adenosine-5'-tetraphosphate behaves differently. The major product is adenosine-5'-monophosphate. We believe that this hydrolysis occurs via a cyclic trimetaphosphate intermediate. PMID- 3033608 TI - A rapid biochemical method for purifying high molecular weight bacterial chromosomal DNA for restriction enzyme analysis. PMID- 3033607 TI - Genomic sequence coding for tunicamycin resistance in yeast. PMID- 3033610 TI - tRNA, cloned human DNA sequences and restriction enzyme recognition sequences. PMID- 3033609 TI - Primary structure of the human beta-adrenergic receptor gene. PMID- 3033611 TI - Restriction enzymes and their isoschizomers. PMID- 3033613 TI - [Congenital infections caused by TORCH agents]. AB - The frequency of transplacental infections varies depending on the pathogen responsible. Of the TORCH infections this paper studies the epidemiology, pathogenesis, clinics, therapy and prophylaxis of infections caused by Rubella, Varicella-Zoster, Cytomegalo and Herpes Simplex Viruses and by Toxoplasma Gondii, Treponema Pallidum. PMID- 3033614 TI - [Mathematical model of monitoring the treatment of severe acidosis with alkalizing agents in acute poisoning]. PMID- 3033615 TI - [Clinical observation of 3 patients with glucagon-secreting pancreatic tumors]. PMID- 3033612 TI - The effect of site-specific methylation on restriction-modification enzymes. PMID- 3033616 TI - High affinity binding of cholecystokinin to small cell lung cancer cells. AB - The binding of 125I-Bolton Hunter-cholecystokinin octapeptide (125I-BH-CCK-8) to small cell lung cancer cell lines was investigated. 125I-BH-CCK-8 bound with high affinity (Kd = 2.4 nM) to an apparent single class of sites (1700/cell) using cell line NCI-H209. Binding was time dependent and the ratio of specific/nonspecific binding was 8/1. Pharmacology studies indicated that gastrin, caerulein, CCK-33 and nonsulfated CCK-8 were potent inhibitors of specific 125I-BH-CCK-8 binding whereas CCK-26-32-NH2 was not. Because CCK receptors are present on small cell lung cancer cells, CCK may function as a regulatory peptide in this disease. PMID- 3033618 TI - Pharmacological examination of cholecystokinin (CCK-8)-induced contractile activity in the rat isolated pylorus. AB - The actions of cholecystokinin (CCK) in the production of a satiety-like state have been suggested to be mediated via receptors for CCK which are located in the pylorus. We investigated the actions of CCK and other pharmacological agents upon the isolated rat pylorus in vitro. We used the change in isometric tension of the tissue preparation (contraction amplitude) as the measure of the effects of the pharmacological agents. Cholecystokinin COOH-terminal octapeptide (CCK-8) was observed to elicit contraction in a dose-dependent manner, with the half-maximal dose (ED50) in the vicinity of 1 nM. Rapid desensitization to CCK was observed. The contraction amplitude was atropine-independent, and was not significantly antagonized by a wide variety of other pharmacological agents. The Na+-channel blocker tetrodotoxin was without effect upon contractile amplitude, as was the K+ channel blocker 4-aminopyridine, except at very high concentrations. Neurotensin, bombesin, and the substance P and bombesin antagonist spantide all elicited contraction in the isolated tissue; neurotensin had a similar potency to CCK-8 and bombesin was 10-15-fold less potent than CCK-8. Unsulfated CCK-8 was at least 170-fold less potent than sulfated CCK-8 and tetragastrin was at least 500-fold less potent than CCK-8. These results suggest that pyloric CCK receptors, which appear to have a pharmacological profile typical of peripheral CCK receptors, may have a physiological role in the peptidergic control of gastric emptying in the rat. PMID- 3033617 TI - Dual effects of (D-Ala2,Met5)-enkephalinamide on CRF and ACTH secretion. AB - An intra-third ventricular administration of (D-Ala2,Met5)-enkephalinamide (DALA) did not elevate plasma ACTH and corticosterone levels in unanesthetized freely moving rats, but intra-third ventricular administration of DALA and methionine (Met)-enkephalin potentiated a mild stress (hanging for 10 or 30 sec)-induced plasma ACTH and corticosterone elevations in unanesthetized freely moving rats. DALA and Met-enkephalin seemed to stimulate CRF release from the median eminence to increase plasma ACTH, as the CRF concentration in the median eminence area was reduced after injection in these stressed rats. When hypothalamic tissues were perifused in vitro, DALA (1-100 ng/ml) reduced the release of CRF. These results suggest that the opiates seem to have a dual effect on the CRF-ACTH system depending on which action overrides the other. PMID- 3033620 TI - [Role of protein kinases and phosphatases in the regulation of glycogen metabolism]. PMID- 3033619 TI - Target size analysis of neurotensin receptors. AB - The neurotensin receptors from rat brain synaptosomal membranes differed in subunit structure from those in rat gastric fundus smooth muscle plasma membranes when studied following photoaffinity labelling or after exposure to cross-linking reagents. However, these two receptors were similar in their recognition properties. In this study we compared the target size of the two receptors by radiation inactivation and observed that the receptors in both tissues had similar target sizes (mean values 103,000 and 108,000 daltons). This suggests that the differences in size observed in biochemical studies may reflect changes occurring during the isolation procedures, or, on the other hand, there might be inherent difference in the subunit structure of these receptors. PMID- 3033621 TI - [The proton-transporting ATPase complex]. PMID- 3033622 TI - [Inositol phospholipids in transforming information in the cells]. PMID- 3033623 TI - Alcohol withdrawal. A comprehensive approach to treatment. PMID- 3033624 TI - Field rickets in turkeys: relationship to vitamin D. AB - Thirty-two outbreaks of leg disorders in turkeys were investigated during 1981 1985. Among them, 22 were characterized by a low percentage of bone ash and were considered as field rickets. Most of the field rickets cases exhibited reduced plasma calcium and inorganic phosphorus. Plasma 25-hydroxyvitamin D3 and intestinal calcium-binding protein were lower in the rachitic than in normal turkeys. These symptoms are typical of rickets resulting from vitamin D deficiency. The vitamin D3 equivalence of a diet that had been fed during field rickets outbreaks was assayed biologically and found to be 111 micrograms/kg diet, about eight times the minimal requirement. In two other cases no symptoms of rickets were observed in turkeys fed diets that had been previously consumed during field rickets outbreaks. The results indicate that in some cases of field rickets there was no involvement of dietary factors and confirm a previous suggestion that field rickets may result from defects in metabolism of vitamin D3, or in its expression. PMID- 3033626 TI - [Bilateral, large cavitary broncho-alveolar carcinoma of a diffuse type]. PMID- 3033625 TI - Studies on the effect of different inhibitors of arachidonic acid metabolism on glyceryltrinitrate-induced relaxation and cGMP elevation in bovine vascular tissue. AB - This study was performed in order to investigate the possible involvement of arachidonic acid metabolites in the mediation of glyceryltrinitrate (GTN)-induced relaxation in isolated bovine mesenteric artery (BMA) and vein (BMV) and in bovine coronary artery (BCA). Concentration-effect curves for GTN were established on the different types of vessels, precontracted with 2.5 microM phenylephrine (BMA) or K+-depolarization (BMV and BCA), in the presence or absence of different inhibitors of the arachidonic acid metabolism. The used inhibitors of arachidonic acid metabolism were: 10 microM quinacrine (a phospholipase A2 inhibitor), 100 microM acetylsalicylic acid, 20 microM indomethacin (cyclooxygenase inhibitors), 3 mM tranylcypromine (inhibitor of PGI2 synthesis), 50 microM nordihydroguairetic acid and 40 microM BW 755C (lipoxygenase inhibitors). In addition, SKF 525A (10 microM) was tested on BMA; this agent is considered to block the cytochrome P450-dependent monooxygenase pathway. The effect on the endogenous levels of cyclic nucleotides after treatment with some of the inhibitors were also measured. Quinacrine and acetylsalicylic acid had no statistical significant effect (P greater than 0.05) on the pD2-value for GTN-induced relaxation in BMA, BMV and BCA. The results obtained with indomethacin were very variable. This drug was almost completely without effect on the GTN induced relaxation in BCA. In BMA a significant potentiation of the relaxant response was obtained (P = 0.002), while in BMV a significant inhibition was seen (P = 0.049).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3033627 TI - [Incidence of hypovitaminosis D3]. PMID- 3033628 TI - [What is the role of Epstein-Barr virus in the pathogenesis of connective tissue diseases?]. AB - In rheumatoid arthritis (RA) abnormalities of T suppressive/cytotoxic responses to Epstein-Barr virus (EBV) have been reported: lack of inhibition of spontaneous proliferation of autologous B lymphocytes infected by EBV, lack of late suppression of immunoglobulin secretion induced in vitro by EBV. Moreover, a cross-reaction between EBNA antigen (Epstein Barr Nuclear Antigen) and a cytoplasmic protein of 62 KD present in the rheumatoid synovitis has been described. In primary Sjogren's syndrome, EBV genome has been observed in the parotid of some patients. The abnormalities of T suppressive/cytotoxic responses observed in rheumatoid arthritis are inconstant and non specific; they are noted in some cases of systemic scleroderma and lupus; they remain for a given patient and could be the marker of a particular subset of connective tissue diseases. The pathophysiological role of EBV infection, especially in rheumatoid arthritis and Sjogren's syndrome, remains to be determined. PMID- 3033629 TI - [Chronic inflammatory polyneuropathy and Behcet's syndrome]. PMID- 3033630 TI - [Isolation and purification of restriction endonuclease SacI from Streptomyces achromogenes ATCC 12767]. AB - A procedure for isolation and purification of restriction endonuclease Sac I from Streptomyces achromogenes ATCC 12767 is proposed. It allows to obtain an electrophoretically homogeneous enzyme preparation with the purification degree 1097 and the enzyme yield by activity 3.7%. The molecular weight of SacI was found to be 52,000 +/- 5,000 D, and isoelectric point 6.2. The enzyme consists of two subunits, which was found by polyacrylamide gel electrophoresis under denaturing conditions. Km and Vmax values were determined for the enzymatic reaction; they are equal to 4.6 X 10(-9) M and 9.19 X 10(-10) M/min, respectively. PMID- 3033631 TI - Purine-metabolizing enzymes in Babesia divergens. AB - Extracts of Babesia divergens were examined for the enzymes which catalyse purine salvage. Adenosine deaminase (EC 3.5.4.4), guanine deaminase (EC 3.5.4.3), inosine phosphorylase (EC 2.4.2.1), purine phosphoribosyltransferases (EC 2.4.2.7, EC 2.4.2.8, EC 2.4.2.22) and nucleoside kinases (EC 2.7.1.15, EC 2.7.1.20, EC 2.7.1.73) were all detected at relatively high activities, whereas nucleotide interconverting enzymes were not detected. Coformycin and 4-amino-5 imidazolecarboxamide were found to be potent inhibitors of adenosine deaminase and guanine deaminase, respectively. The results suggest that B. divergens is capable of synthesizing purine nucleotides via two routes, one involving purine phosphoribosyltransferases and the other employing nucleoside kinases. PMID- 3033632 TI - Coexisting pancreatic and breast adenocarcinomas: is there an association? AB - Two patients are reported with coexisting adenocarcinomas of pancreas and breast. One represents synchronous neoplasms, the other metachronous tumors diagnosed 7 years apart. The significance of these findings is discussed, including possible carcinogenic dietary factors common to both neoplasms. The potential problems associated with the histologic diagnosis of such coexisting tumors are stressed. PMID- 3033634 TI - Sequences from a prokaryotic genome or the mouse dihydrofolate reductase gene can restore the import of a truncated precursor protein into yeast mitochondria. AB - Sequences that are capable of restoring mitochondrial targeting to a truncated yeast cytochrome c oxidase subunit IV presequence are encoded within the genome of Escherichia coli and within the gene for a higher eukaryotic cytosolic protein, mouse dihydrofolate reductase. These sequences, which resemble authentic presequences in their overall amino acid composition and degree of hydrophobicity, are rather frequent; greater than 2.7% of clones generated from E. coli DNA and greater than 5% of clones from the dihydrofolate reductase gene were functional in our screening system. These results suggest that, during evolution, mitochondrial precursor proteins could arise as a result of DNA rearrangements that place potential mitochondrial presequences at the amino terminus of existing open reading frames. Primitive eukaryotic cells may have used this mechanism to target proteins to their endosymbiotic protomitochondria. PMID- 3033633 TI - Cloning and DNA sequence of the mercuric- and organomercurial-resistance determinants of plasmid pDU1358. AB - The broad-spectrum mercurial-resistance plasmid pDU1358 was analyzed by cloning the resistance determinants and preparing a physical and genetic map of a 45 kilobase (kb) region of the plasmid that contains two separate mercurial resistance operons that mapped about 20 kb apart. One encoded narrow-spectrum mercurial resistance to Hg2+ and a few organomercurials; the other specified broad-spectrum resistance to phenylmercury and additional organomercurials. Each determinant governed mercurial transport functions. Southern DNA X DNA hybridization experiments using gene-specific probes from the plasmid R100 mer operon indicated close homology with the R100 determinant. The 2153 base pairs of the promoter-distal part of the broad-spectrum Hg2+-resistance operon of pDU1358 were sequenced. This region included the 3'-terminal part of the merA gene, merD, unidentified reading frame URF1, and a part of URF2 homologous to previously sequenced determinants of plasmid R100. Between the merA and merD genes, an open reading frame encoding a 212 amino acid polypeptide was identified as the merB gene that determines the enzyme organomercurial lyase that cleaves the C--Hg bond of phenylmercury. PMID- 3033635 TI - Properties of the duplex DNA-dependent ATPase activity of Escherichia coli RecA protein and its role in branch migration. AB - We have investigated the double-stranded DNA (dsDNA)-dependent ATPase activity of recA protein. This activity is distinguished from the single-stranded DNA (ssDNA) dependent ATPase activity by the presence of a pronounced lag time before the onset of steady-state ATP hydrolysis. During the lag phase there is little ATP hydrolysis. The duration of the lag phase, referred to as the lag time, is found to increase with the thermal stability of the dsDNA substrate. Increasing either the MgCl2 or NaCl concentration increases the lag time, whereas increasing the temperature decreases the lag time. The lag time shows little dependence on recA protein concentration but is strongly dependent on ATP concentration. After the lag phase, a steady-state ATP hydrolysis rate is achieved that approaches the rate observed with ssDNA. The steady-state phase of the reaction is proportional to the concentration of recA protein-DNA complex and shows saturation behavior at approximately equal to 5 +/- 1 base pairs per recA protein monomer. These results suggest that the lag phase represents a rate-limiting step in the dsDNA-dependent ATP hydrolysis reaction that requires a structural transition in the dsDNA and that involves a ternary complex of ATP, recA protein, and DNA. We propose that this transition involves the transient denaturation of the dsDNA to form regions of ssDNA. Elsewhere we demonstrate that the dsDNA-dependent ATPase activity is proportional to the rate of recA protein-catalyzed branch migration. We suggest that this activity is responsible for a polar polymerization that drives the branch migration reaction. PMID- 3033636 TI - Insulin stimulates phosphorylation of a 120-kDa glycoprotein substrate (pp120) for the receptor-associated protein kinase in intact H-35 hepatoma cells. AB - The insulin receptor possesses protein kinase activity, which may play a role in mediating insulin action. Recently, we have identified a glycoprotein (pp120) in rat liver plasma membranes that is phosphorylated by the solubilized insulin receptor in a cell-free system. We now report that insulin stimulates phosphorylation of pp120 in intact H-35 cells. H-35 cells were preloaded with [32P]orthophosphate to label the intracellular ATP pool. Insulin caused a 10-fold increase in the phosphorylation of its receptor and a 2-fold increase in phosphorylation of pp120 (P less than 0.001). The time course of insulin's stimulation of pp120 closely paralleled that of insulin receptor phosphorylation over the time period investigated (15-45 min). This effect had the specificity corresponding to the insulin receptor. Epidermal growth factor was inactive, and insulin-like growth factor I had approximately equal to 1% the potency of insulin in this regard. Insulin increased 32P incorporation into pp120 in a linkage that was stable to alkaline hydrolysis, as would be expected for tyrosine-specific phosphorylation. Direct phosphoamino acid analysis confirmed that insulin increased 32P incorporation into phosphotyrosine residues in pp120. PMID- 3033637 TI - Cotranslational insertion of selenocysteine into formate dehydrogenase from Escherichia coli directed by a UGA codon. AB - The structural gene (fdhF) for the 80-kDa selenopolypeptide of formate dehydrogenase (formate:benzyl viologen oxidoreductase, EC 1.2.--.--) from Escherichia coli contains an in-frame UGA codon at amino acid position 140 that is translated. Translation of gene fusions between N-terminal parts of fdhF with lacZ depends on the availability of selenium in the medium when the hybrid gene contains the UGA codon; it is independent of the presence of selenium when an fdhF portion upstream of the UGA position is fused to lacZ. Transcription does not require the presence of selenium in either case. By localized mutagenesis, the UGA codon was converted into serine (UCA) and cysteine (UGC and UGU) codons. Each mutation relieved the selenium dependency of fdhF mRNA translation. Selenium incorporation was completely abolished in the case of the UCA insertion and was reduced to about 10% when the UGA was replaced by a cysteine codon. Insertion of UCA yielded an inactive fdhF gene product, while insertion of UGC and UGU resulted in polypeptides with lowered activities as components in the system formerly known as formate hydrogenlyase. Altogether the results indicate that the UGA codon at position 140 directs the cotranslational insertion of selenocysteine into the fdhF polypeptide chain. PMID- 3033638 TI - Aberrant splicing events that are induced by proviral integration: implications for myb oncogene activation. AB - Activation of the mouse c-myb oncogene in Abelson virus-induced plasmacytoid lymphosarcomas was studied using cDNA cloning and nucleotide sequence analysis. The results presented here show that viral integration in the myb locus generates splicing errors at the 5' and 3' regions. Viral integration results in transcriptional initiation within the viral long terminal repeat and generation of a chimeric mRNA that lacks the first three coding exons. The alterations at the 3' end are caused by an aberrant splicing event in which additional splice donor and -acceptor sequences within intronic sequences are used to splice an additional 363 nucleotides into the myb transcripts. The resulting insertion of 121 amino acids is in a region of the protein where other activated forms of the myb gene product have deletions. These results suggest that alterations in the 3' end of the myb gene play a crucial role in the activation of this gene. PMID- 3033639 TI - Role of DNA topoisomerase I in the transcription of supercoiled rRNA gene. AB - The fraction DE-B obtained by fractionating an extract from rat mammary adenocarcinoma cells on a DEAE-Sephadex column was used for transcribing linear and supercoiled rRNA gene (rDNA). This fraction, which is known to contain RNA polymerase I and essential transcription factors, also contains DNA topoisomerase I activity. Inhibition of this topoisomerase activity by the selective inhibitor camptothecin markedly diminished transcription of supercoiled rDNA, and at a concentration of 150 microM, camptothecin almost completely inhibited DNA topoisomerase I activity and supercoiled rDNA transcription. Addition of exogenous calf thymus DNA topoisomerase I to the sample containing the drug restored the ability of the extract to transcribe supercoiled rDNA. Camptothecin, even at a concentration of 500 microM, had no significant effect on the transcription of linear rDNA. These studies show that relaxation of supercoiled rDNA by DNA topoisomerase I is essential for its transcription. The preferential inhibition of rRNA synthesis in vivo following treatment with camptothecin is probably due to selective camptothecin inhibition of DNA topoisomerase I activity. PMID- 3033640 TI - RNA from an immediate early region of the type 1 herpes simplex virus genome is present in the trigeminal ganglia of latently infected mice. AB - Transcription of the type 1 herpes simplex virus (HSV-1) genome in trigeminal ganglia of latently infected mice was studied using in situ hybridization. Probes representative of each temporal gene class were used to determine the regions of the genome that encode the transcripts present in latently infected cells. Probes encoding HSV-1 sequences of the five immediate early genes and representative early (thymidine kinase), early-late (major capsid protein), and late (glycoprotein C) genes were used in these experiments. Of the probes tested, only those encoding the immediate early gene product infected-cell polypeptide (ICP) 0 hybridized to RNA in latently infected tissues. Probes containing the other immediate early genes (ICP4, ICP22, ICP27, and ICP47) and the representative early, early-late, and late genes did not hybridize. Two probes covering approximately equal to 30% of the HSV-1 genome and encoding over 20 early and late transcripts also did not hybridize to RNA in latently infected tissues. These results, with probes spanning greater than 60% of the HSV-1 genome, suggest that transcription of the HSV-1 genome is restricted to one region in latently infected mouse trigeminal ganglia. PMID- 3033641 TI - The very late antigen family of heterodimers is part of a superfamily of molecules involved in adhesion and embryogenesis. AB - The very late antigen (VLA) protein family contains at least five related heterodimers, including a fibronectin receptor structure, and probably other cell substrate adhesion receptors. These cell-surface VLA proteins were immunopurified from human placenta (VLA-1, VLA-3, and VLA-5), platelets (VLA-2), and Molt-4 cells (VLA-4) using a series of monoclonal antibody-Sepharose immunoaffinity columns. After further purification by gel electrophoresis, the N-terminal amino acid sequence for each of the five VLA alpha subunits was determined. In the first 14 positions, the five VLA alpha subunits showed an average of 42% homology to each other, rising to 59% including conservative amino acid substitutions. In addition, the alpha subunits from the LFA-1, Mac-1 (CR-3), and p150,95 family of heterodimers, the vitronectin receptor-platelet GPIIb/IIIa family, and a position specific (PS) antigen important in Drosophila embryogenesis each showed average homologies of 31-40% to individual VLA alpha sequences and 46-52% homology to VLA alpha subunits including conservative substitutions. Taken together, these results suggest that the VLA proteins, the LFA-1, Mac-1, and p150,95 family, the GPIIb/IIIa, vitronectin receptor family, and the Drosophila PS antigens have evolved as four subgroups in a highly conserved supergene family of receptors involved in fundamentally important functions, such as cell adhesion, migration, and embryogenesis. PMID- 3033642 TI - A high molecular weight component of the human tumor necrosis factor receptor is associated with cytotoxicity. AB - We compared the molecular structure of the receptor to human recombinant tumor necrosis factor (HurTNF) on cells of different tissue origin that differ in their response to one of the known activities of TNF. We studied tumor cell lines that respond to the cytotoxic action of TNF and resistant variants that bind TNF, normal cell lines that are stimulated to proliferate by TNF and those that are not affected by TNF, and peripheral blood granulocytes whose activation is also augmented by TNF. Using 125I-labeled HurTNF, we found that it bound mainly to four cellular polypeptides (138, 90, 75, and 54 kDa), three of which were found in every cell type examined and one (138 kDa) that was observed only in a human breast carcinoma cell line (MCF-7) that is highly responsive to the cytotoxic action of TNF. The 138-kDa polypeptide was not found in resistant variants of MCF 7 that bind TNF. In contrast to the other polypeptides, the 138-kDa protein was detected 30 min after incubation at 4 degrees C, as compared to 5 min. Scatchard analysis and cross-linking data suggest a model for the TNF receptor structure whereby the receptor is composed of noncovalently linked membrane-bound polypeptides that bind TNF with high affinity (Kd, 0.05-0.8 X 10(-9) M) with the 138-kDa protein being the least abundant and/or even absent in most cells. PMID- 3033643 TI - Structure and expression of human dihydropteridine reductase. AB - Dihydropteridine reductase (DHPR; EC 1.6.99.7) catalyzes the NADH-mediated reduction of quinonoid dihydrobiopterin and is an essential component of the pterin-dependent aromatic amino acid hydroxylating systems. A cDNA for human DHPR was isolated from a human liver cDNA library in the vector lambda gt11 using a monospecific antibody against sheep DHPR. The nucleic acid sequence and amino acid sequence of human DHPR were determined from a full-length clone. A 112 amino acid sequence of sheep DHPR was obtained by sequencing purified sheep DHPR. This sequence is highly homologous to the predicted amino acid sequence of the human protein. Gene transfer of the recombinant human DHPR into COS cells leads to expression of DHPR enzymatic activity. These results indicate that the cDNA clone identified by antibody screening is an authentic and full-length cDNA for human DHPR. PMID- 3033644 TI - Expression of retrovirally transduced genes in primary cultures of adult rat hepatocytes. AB - Differentiated primary rat hepatocyte cultures have been infected with retroviral vectors expressing human hypoxanthine/guanine phosphoribosyltransferase or the transposon Tn5 neomycin-resistance gene. Expression of the markers was detected only after infection of the cells during a short period of cell replication and transient dedifferentiation from days 1 to 5 of culture. Provirus integrated during that period remains fully expressed during the entire subsequent stationary period of culture up to at least 25 days. Selection with the neomycin analogue G418 of cells infected with the neomycin vector led to the appearance of cells with hepatocyte morphology in which newly synthesized albumin was detectable by immunoprecipitation, indicating successful infection of hepatocytes. PMID- 3033645 TI - Biosynthesis, glycosylation, and partial N-terminal amino acid sequence of the T cell-activating protein TAP. AB - We have characterized the TAP molecule, an Ly-6 linked T-cell-activating glycoprotein. The three TAP bands that are precipitated from metabolically labeled cells display a common migration pattern in isoelectric focusing/NaDodSO4/PAGE gels and have common N-terminal sequences. This sequence is rich in cysteine and is homologous to that previously reported for the Ly-6.1E antigen. We, therefore, compared TAP and Ly-6.1E biochemically and found them to be structurally distinct. Given the role of TAP in T-cell activation, we further studied whether the molecule was phosphorylated. We have not found evidence for phosphorylation of the TAP protein. The carbohydrates present on the TAP molecule are resistant to peptide N-glycosidase F in vitro and tunicamycin in vivo. The upper band of the TAP triplet is susceptible to treatment with trifluoromethanesulfonic acid and thus seems to be of the O-linked rather than of the N-linked variety. The biosynthetic processing of TAP was studied in pulse chase experiments. The middle band of the TAP triplet appears to be the earliest detectable species. Its conversion to the O-linked high molecular weight species can be blocked by monensin. PMID- 3033646 TI - 1 alpha,25-Dihydroxyvitamin D3 inhibits gamma-interferon synthesis by normal human peripheral blood lymphocytes. AB - 1 alpha,25-Dihydroxyvitamin D3 [1,25-(OH)2D3], the biologically active metabolite of vitamin D3, inhibited synthesis of gamma-interferon (IFN-gamma) by phytohemagglutinin-activated peripheral blood lymphocytes (PBLs). A significant reduction of IFN-gamma protein levels in PBL culture medium was achieved with a physiologic 1,25-(OH)2D3 concentration (0.1 nM). 1,25-(OH)2D3 also inhibited accumulation of IFN-gamma mRNA in activated PBLs in a dose-dependent fashion. The ability of 1,25-(OH)2D3 to modulate IFN-gamma protein synthesis was unaltered in the presence of high concentrations of recombinant human interleukin 2. The suppression of IFN-gamma synthesis by PBLs was specific for 1,25-(OH)2D3; the potencies of other vitamin D3 metabolites were correlated with their affinities for the cellular 1,25-(OH)2D3 receptor. The time course of 1,25-(OH)2D3 receptor expression in phytohemagglutinin-activated PBLs was correlated with the time course of 1,25-(OH)2D3-mediated inhibition of IFN-gamma synthesis. In selected experiments, T-lymphocyte-enriched cell preparations were utilized. In these experiments, 1,25-(OH)2D3 was equally active as in PBL preparations. Finally, we examined the effects of 1,25-(OH)2D3 on the constitutive IFN-gamma production by two human T-lymphocyte lines transformed by human T-lymphotropic virus type I. The cell lines were established from a normal donor (cell line S-LB1) and from a patient with vitamin D-dependent rickets type 2 (cell line Ab-VDR). IFN-gamma synthesis by S-LB1 cells was inhibited in a dose-dependent fashion by 1,25 (OH)2D3, whereas IFN-gamma synthesis by Ab-VDR cells was not altered by 1,25 (OH)2D3. The data presented in this study provide further evidence for a role of 1,25-(OH)2D3 in immunoregulation. PMID- 3033647 TI - Glucagon-like peptide I stimulates insulin gene expression and increases cyclic AMP levels in a rat islet cell line. AB - Insulin secretion is controlled by a complex set of factors. Although blood glucose levels serve as the major stimulus of insulin secretion in mammals, insulin release is also modulated by amino acids, catecholamines, glucagon, and other, intestinal hormones. The identification of factors that modulate insulin production has engendered much interest because of their potential importance in the altered dynamics of insulin secretion in response to glucose characteristic of maturity-onset diabetes mellitus. Decoding of the glucagon gene has uncovered two additional glucagon-like peptides encoded in proglucagon, the polypeptide precursor of glucagon. One of these peptides, glucagon-like peptide I, is processed from proglucagon in two forms, of 31 and 37 amino acids. We report that the smaller of the two glucagon-like peptides potently increases cAMP levels, insulin mRNA transcripts, and insulin release in cultured rat insulinoma cells. These results indicate that glucagon-like peptide I may be a physiologic modulator of insulin gene expression. PMID- 3033648 TI - Monoclonal antibodies to the human insulin receptor that activate glucose transport but not insulin receptor kinase activity. AB - Three mouse monoclonal antibodies were produced that reacted with the alpha subunit of the human insulin receptor. All three both immunoprecipitated 125I labeled insulin receptors from IM-9 lymphocytes and competitively inhibited 125I labeled insulin binding to its receptor. Unlike insulin, the antibodies failed to stimulate receptor autophosphorylation in both intact IM-9 lymphocytes and purified human placental insulin receptors. Moreover, unlike insulin, the antibodies failed to stimulate receptor-mediated phosphorylation of exogenous substrates. However, like insulin, two of the three antibodies stimulated glucose transport in isolated human adipocytes. One antibody, on a molar basis, was as potent as insulin. These studies indicate, therefore, that monoclonal antibodies to the insulin receptor can mimic a major function of insulin without activating receptor kinase activity. They also raise the possibility that certain actions of insulin such as stimulation of glucose transport may not require the activation of receptor kinase activity. PMID- 3033649 TI - Epstein-Barr virus nuclear antigen 2 specifically induces expression of the B cell activation antigen CD23. AB - Epstein-Barr virus (EBV) infection of EBV-negative Burkitt lymphoma (BL) cells induces some changes similar to those seen in normal B lymphocytes that have been growth transformed by EBV. The role of individual EBV genes in this process was evaluated by introducing each of the viral genes that are normally expressed in EBV growth-transformed and latently infected lymphoblasts into an EBV-negative BL cell line, using recombinant retrovirus-mediated transfer. Clones of cells were derived that stably express the EBV nuclear antigen 1 (EBNA-1), EBNA-2, EBNA-3, EBNA-leader protein, or EBV latent membrane protein (LMP). These were compared with control clones infected with the retrovirus vector. All 10 clones converted to EBNA-2 expression differed from control clones or clones expressing other EBV proteins by growth in tight clumps and by markedly increased expression of one particular surface marker of B-cell activation, CD23. Other activation antigens were unaffected by EBNA-2 expression, as were markers already expressed on the parent BL cell line, including BL markers (cALLA and BLA), proliferation markers (transferrin receptor and BK19.9), and cell adhesion-related molecules (LFA-1 and LFA-3). Increased CD23 expression in cells expressing EBNA-2 was apparent from monoclonal anti-CD23 antibody binding to the cell surface, from immunoprecipitation of the 45-kDa and 90-kDa CD23 proteins with monoclonal antibody, and from RNA blots probed with labeled CD23 DNA. The results indicate that EBNA-2 is a specific direct or indirect trans-activator of CD23. This establishes a link between an EBV gene and cell gene expression. Since CD23 has been implicated in the transduction of B-cell growth signals, its specific induction by EBNA-2 could be important in EBV induction of B-lymphocyte transformation. PMID- 3033650 TI - Identification and purification of an irreversible presynaptic neurotoxin from the venom of the spider Hololena curta. AB - A search for potent toxins that inhibit neuronal calcium channels in Drosophila melanogaster has resulted in the identification of a presynaptic neurotoxin from the venom of the hunting spider Hololena curta. Using Drosophila neuromuscular junction as an assay, presynaptic inhibitory activity was purified using gel filtration and reverse-phase HPLC. Data from gel electrophoresis indicate that the toxin is composed of two different subunits of Mr 7000 and 9000. At nanomolar concentrations the toxin produced a complete and long-lasting inhibition of synaptic transmission at the Drosophila larval neuromuscular junction without affecting the amplitudes of the spontaneously occurring miniature junction potentials. The block of transmission produced by the toxin was observed even during the direct depolarization of the motor nerve terminal. These physiological results indicate that the terminal is the site of action for the toxin. Indirect evidence using abnormally excitable Drosophila mutants suggests that the toxin is inhibiting transmitter release by altering the electrical properties of the nerve terminal rather than by interfering with nonelectrical events that may occur subsequent to calcium influx. All of the actions of the Hololena toxin can be explained by a specific and direct effect on presynaptic calcium channels in Drosophila motor neurons. PMID- 3033651 TI - Cloning and heterologous expression of glycosidase genes from Saccharomyces cerevisiae. AB - Genomic clones were isolated that code for three glycosidases proposed to be involved in the catabolism of cell wall components in Saccharomyces cerevisiae. alpha-Mannosidase (AMS1), exoglucanase (BGL1), and endochitinase (CTS1) genes were isolated with the aid of filter assays based on the hydrolysis of 4 methylumbelliferyl glycosides, which permitted the in situ monitoring of these glycosidase activities in yeast colonies. Uracil prototrophs resulting from transformation with a multicopy YEp24 yeast genomic library were screened, leading to the identification of transformants possessing high levels of glycosidase activity. Restriction maps of plasmids from multiple isolates were used to localize glycosidase-overproduction genes, which were subcloned into a Schizosaccharomyces pombe/S. cerevisiae shuttle vector. Transformation of Sch. pombe with BGL1 and CTS1 subclones resulted in the appearance of these activities in this organism, and an AMS1 plasmid caused a 2-fold increase in endogenous alpha-mannosidase levels. Insertion of the marker gene LEU2 into putative AMS1 sequences disrupted plasmid-encoded alpha-mannosidase overproduction. S. cerevisiae strains that incorporated a restriction fragment containing ams1::LEU2 into their chromosomal DNA by homologous recombination expressed no detectable alpha-mannosidase activity in either the haploid or homozygous recessive diploid states, whereas heterozygous and wild-type cells exhibited levels proportional to AMS1 gene dosage. No readily apparent phenotype was associated with the alpha mannosidase deficiency; however, labeling experiments utilizing [2-3H]mannose suggest that alpha-mannosidase may function in mannan turnover. PMID- 3033652 TI - Coordinate regulation of stromelysin and collagenase genes determined with cDNA probes. AB - Secreted proteinases are required for tumor metastasis, angiogenesis, and tissue remodeling during wound healing and embryonic growth. Thus, the regulation of the genes of secreted proteinases may serve as an interesting model for growth controlled genes in general. We studied the genes of the secreted proteinases stromelysin and collagenase by using molecularly cloned cDNAs from each proteinase. Stromelysin cDNA was cloned by differential screening of a total cDNA library from rabbit synovial cells treated with phorbol 12-myristate 13-acetate, which yielded a clone of 1.2 kilobase pairs; collagenase cDNA was obtained by cloning reverse transcripts of anti-collagenase-immunoadsorbed polysomal mRNA, which yielded a clone of 0.8 kilobase pairs. Stromelysin and collagenase mRNA species of 2.2 and 2.4 kilobases, respectively, were detected on hybridization blots of RNA from phorbol 12-myristate 13-acetate-treated but not untreated rabbit synovial cells. Expression of stromelysin mRNA was also induced in rabbit alveolar macrophages and rabbit brain capillary endothelial cells treated with phorbol 12-myristate 13-acetate. Stromelysin and collagenase mRNA were both induced by phorbol 12-myristate 13-acetate and cytochalasin B at a constant ratio of the two gene products; this suggests coordinate regulation. The fact that induction was blocked after inhibition of protein synthesis by cycloheximide implicates an indirect signal transduction pathway that requires new protein synthesis. PMID- 3033653 TI - Evolutionary relationships within the human rhinovirus genus: comparison of serotypes 89, 2, and 14. AB - The complete nucleotide sequence of the genome of human rhinovirus type 89 was determined from the cDNA that had been cloned into Escherichia coli. The genome is 7152 nucleotides long and contains a single large open reading frame of 2164 codons. Translation commences at position 619 and ends 42 nucleotides before the poly(A) tract. The positions of three proteolytic cleavage sites in the polyprotein were determined by N-terminal amino acid sequencing of the capsid proteins; the remainder were predicted from comparisons with other picornaviruses. Extensive similarity between the derived amino acid sequences of human rhinovirus types 89 and 2 was found, whereas the similarity between human rhinovirus types 89 and 14 was considerably less. It is apparent that human rhinoviruses may be more closely related than has been previously thought. PMID- 3033654 TI - 24- and 26-homo-1,25-dihydroxyvitamin D3: preferential activity in inducing differentiation of human leukemia cells HL-60 in vitro. AB - 1,25-Dihydroxyvitamin D3, the hormonal form of vitamin D3, promotes the differentiation of HL-60 human promyelocytic leukemia cells into monocytes. Differentiation changes include the induction of phagocytosis, the initiation of nitroblue tetrazolium-reducing activity, and the appearance of nonspecific acid esterase. We have found that the 24-homo- and 26-homo-1,25-dihydroxyvitamin D3 and their delta 22 analogues are 10-fold more potent than 1,25-dihydroxyvitamin D3 in inducing differentiation of HL-60 cells in vitro. In vivo, these analogues show activity similar to 1,25-dihydroxyvitamin D3 in stimulating intestinal calcium transport in vitamin D-deficient rats. The 24-homoanalogues are significantly less active, whereas the 26-homo derivatives are more active than the natural hormone in mobilizing calcium from bone. This unusual activity pattern cannot be explained on the basis of the affinity of these analogues for the 1,25-dihydroxyvitamin D3 intracellular receptor: both 24-homo- and 26-homo 1,25-dihydroxyvitamin D3 have the same effectiveness as 1,25-dihydroxyvitamin D3 in displacing the tritiated hormone from its receptor in rat intestine or HL-60 cells. These analogues of 1,25-dihydroxyvitamin D3 may be of some interest as possible therapeutic substances, or as tools in understanding the action of 1,25 dihydroxyvitamin D3 in inducing differentiation. PMID- 3033656 TI - Effects of DNA supercoiling on the topological properties of nucleosomes. AB - In the nucleosome core particle, at least 145 base pairs of DNA are bound to the histone octamer in a superhelical conformation. We have asked what effect the presence of these particles has on the ability of DNA gyrase to supercoil DNA. Synthetic minichromosomes, constructed by reconstituting complexes of core histones with the closed circular plasmid pBR322, were treated with various amounts of DNA gyrase. We have found that the maximum level of supercoiling that is attainable is nearly identical for protein-free plasmids and for plasmids half saturated with core histones, even though supercoiling does not result in a loss of histones from the complex. It appears that, at sufficiently high levels of supercoiling, the core particle is disrupted in such a way that the DNA bound to histones is no longer constrained. PMID- 3033655 TI - Characterization of a Na+/H+ antiporter gene of Escherichia coli. AB - A mutant of Escherichia coli with increased Na+/H+ antiport activity was isolated and found by other workers to harbor two mutations [Niiya, S., Yamasaki, K., Wilson, T.H. & Tsuchiya, T. (1982) J. Biol. Chem. 257, 8902-8906]. The mutation that leads to increased Na+/H+ antiport (antup) has now been separated and mapped. antup maps in the vicinity of 0.5 min on the E. coli map, and the presence of this mutation alone in an isogenic pair raises antiport activity (Vmax) by approximately 4-fold. We also characterized cells hearing plasmids containing fragments of a 15-kilobase-pair segment of DNA between carA and dnaJ. A wild-type gene located within 2 kilobase pairs counterclockwise of rpsT increases the Na+/H+ antiport activity when present in multiple copies. PMID- 3033657 TI - Expression of a synthetic gene encoding human insulin-like growth factor I in cultured mouse fibroblasts. AB - A synthetic gene encoding human insulin-like growth factor I (hIGF-I) was assembled and inserted into an expression vector containing the cytomegalovirus immediate early (CMV-IE) transcriptional regulatory region and portions of the bovine growth hormone gene. The recombinant plasmid encodes a 97 amino acid fusion protein containing the first 27 amino acids of the bovine growth hormone precursor and the 70 amino acids of hIGF-I. This plasmid, when transiently introduced into cultured mouse fibroblasts, directs synthesis of the fusion protein, subsequent proteolytic removal of the bovine growth hormone signal peptide, and secretion of hIGF-I into the culture medium. Conditioned medium from transfected cells inhibits binding of 125I-labeled IGF-I to type I IGF receptors on human placental membranes and to acid-stable human serum carrier proteins. The recombinant hIGF-I produced is biologically active, as monitored by the stimulation of DNA synthesis in vascular smooth muscle cells. PMID- 3033658 TI - Purification of a phosphatidylinositol-glycan-specific phospholipase C from liver plasma membranes: a possible target of insulin action. AB - Insulin stimulates the hydrolysis of a phosphatidylinositol-glycan, resulting in the generation of two related inositol phosphate-glycan enzyme modulators and diacylglycerol. A phosphatidylinositol-glycan-specific phospholipase C that catalyzed this reaction was found in the plasma-membrane fraction of rat liver. The enzymatic activity was measured by the release of diacylglycerol from the glycosylated-phosphatidylinositol-membrane-anchored variant surface glycoproteins of African trypanosomes. The enzyme also catalyzed the production of an inositol phosphate-glycan from an insulin-sensitive phosphatidylinositol-glycan precursor. The enzyme was solubilized with neutral nonionic detergent and purified to near homogeneity by anion-exchange chromatography on DEAE-cellulose, followed by hydrophobic chromatography on butyl-agarose. The resulting enzyme preparation was purified approximately 20,000-fold from whole liver and exhibited one major silver-stained band of Mr 52,000 on NaDodSO4/PAGE. Gel permeation chromatography of the purified activity revealed a Stokes radius of 35 A and an apparent molecular weight of 62,000, suggesting that the enzyme was tightly associated with lipid or detergent but existed as a monomer in its active form. The enzyme was specific for glycosylated phosphatidylinositol; no hydrolysis of phosphatidylinositol, phosphatidylinositol 4,5-bisphosphate, or phosphatidylcholine was observed. The enzyme was calcium independent and thiol activated. These data suggest a role for the phosphatidylinositol-glycan-specific phospholipase C as an effector for some of the metabolic actions of insulin. PMID- 3033659 TI - Hepatitis B virus (HBV) particles are produced in a cell culture system by transient expression of transfected HBV DNA. AB - An in vitro system for the production of hepatitis B virus (HBV) particles was established by the transient expression of transfected HBV DNA using a human hepatocellular carcinoma cell, HuH-7, as a recipient. The 3.6- and 2.2-kilobase transcripts observed were similar to those in virus-infected liver cells. Both transcripts revealed the microheterogeneity of their 5' ends. The formation of virus-related particles subsequent to the RNA transcription was demonstrated. The core particles observed in the cytoplasm and the virus particles secreted in the culture medium contained the replicative intermediates of HBV DNA and banded at densities of 1.35-1.36 g/cm3 and 1.22-1.24 g/cm3, respectively. Furthermore, the in vitro mutagenesis of the template HBV DNA demonstrated that the P gene as well as the C gene products were essential for the production of HBV particles. PMID- 3033660 TI - Escherichia coli DnaX product, the tau subunit of DNA polymerase III, is a multifunctional protein with single-stranded DNA-dependent ATPase activity. AB - The dnaZX gene of Escherichia coli directs the synthesis of two proteins, DnaZ and DnaX. These products are confirmed as the gamma and tau subunits of DNA polymerase III because antibody to a synthetic peptide present in both the DnaZ and DnaX proteins reacts also with the gamma and tau subunits of holoenzyme. To characterize biochemically the tau subunit, for which there has been no activity assay, the dnaZX gene was fused to the beta-galactosidase gene to encode a fusion product in which the 20 C-terminal amino acids of the DnaX protein (tau) were replaced by beta-galactosidase lacking only 7 N-terminal amino acids. The 185-kDa fusion protein, which retained beta-galactosidase activity, was overproduced to the level of about 5% of the soluble cellular protein by placing the gene fusion under control of the tac promoter and Shine-Dalgarno sequence. The fusion protein was isolated in one step by affinity chromatography on p-aminobenzyl 1-thio-beta D-galactopyranoside-agarose. The purified fusion protein also had ATPase (and dATPase) activity that was dependent on single-stranded DNA. This activity copurified with the beta-galactosidase activity not only through the affinity column but also through a subsequent gel filtration. We conclude that the DnaX protein function involves binding to single-stranded DNA and hydrolysis of ATP or dATP, in addition to binding to other DNA polymerase III holoenzyme components, increasing the processivity of the core enzyme, and serving as a substrate for the production of the gamma subunit. PMID- 3033661 TI - Replacement of the cytoplasmic domain alters sorting of a viral glycoprotein in polarized cells. AB - The envelope glycoprotein (G protein) of vesicular stomatitis virus (VSV) is transported to the basolateral plasma membrane of polarized epithelial cells, whereas the hemagglutinin glycoprotein (HA protein) of influenza virus is transported to the apical plasma membrane. To determine if the cytoplasmic domain of VSV G protein might be important in directing G protein to the basolateral membrane, we derived polarized Madin-Darby canine kidney cell lines expressing G protein or G protein with its normal cytoplasmic domain replaced with the cytoplasmic domain from an influenza HA protein (GHA protein). Indirect immunofluorescence microscopy showed that G protein was present primarily on basolateral surfaces, whereas the GHA protein was present on the apical and basolateral membranes. These results suggest that the cytoplasmic domain can be an important determinant directing polarized expression of an integral membrane protein. PMID- 3033663 TI - Rous sarcoma virus is integrated but not expressed in chicken early embryonic cells. AB - We have developed a protocol that allows us to infect chicken early embryonic (CEE) cells with high efficiency. This was achieved by exposing the CEE cells to a semicontinuous dose of Rous sarcoma virus (RSV) for a period of 20 hr. Southern blot analysis indicated that an average of one proviral copy is integrated per embryonic cell. However, there was no production of infectious viral particles by the cells containing the proviral genome, although low levels of full-length genomic RNA could be detected by RNA transfer blot analysis. These low RNA levels contrast with the 100- to 1000-fold higher levels found in RSV-infected chicken embryo fibroblasts. We conclude that in cells derived from pregastrulating chicken embryos, RSV DNA is integrated into the cell genome but fails to be expressed in an efficient manner. These primary cells can therefore be used to identify factors involved in regulation of retroviral gene expression in normal cells. Such factors may also be instrumental in elucidating basic mechanisms involved in gene regulation during early development in higher vertebrates. PMID- 3033662 TI - Limited cleavage of cellular fibronectin by plasminogen activator purified from transformed cells. AB - The substrate specificity and direct catalytic activity of plasminogen activator (PA) was examined under conditions where its natural substrate, plasminogen, was missing or inhibited. PA, purified from cultures of transformed chicken fibroblasts, was incubated with purified preparations of potential substrates. The adhesive glycoprotein fibronectin, isolated from normal chicken fibroblast extracellular matrix, underwent limited but specific cleavage by PA in the absence of plasminogen. Analysis of the cleavage products by polyacrylamide gels under both reducing and nonreducing conditions indicated that PA-mediated cleavage occurred near the carboxyl terminus of fibronectin but on the amino terminal side of the interchain disulfide bridge, thus disrupting the native dimeric fibronectin molecule. Under the identical conditions, chicken ovalbumin was not cleaved while the established substrate, chicken plasminogen, was extensively converted to plasmin. A monoclonal antibody, directed against avian PA and shown to inhibit plasminogen-free, cell-mediated matrix degradation, specifically inhibited the fibronectin cleavage. A human PA, urokinase, also cleaved fibronectin under plasminogen-free conditions yielding a limited number of high molecular weight cleavage products. PMID- 3033664 TI - Tissue-specific expression of the human type II collagen gene in mice. AB - Type II collagen is crucial to the development of form in vertebrates as it is the major protein of cartilage. To study the factors regulating its expression we introduced a cosmid containing the human type II collagen gene, including 4.5 kilobases of 5' and 2.2 kilobases of 3' flanking DNA, into embryonic stem cells in vitro. The transformed cells contribute to all tissues in chimeric mice allowing the expression of the exogenous gene to be studied in vivo. Human type II collagen mRNA is restricted to tissues showing transcription from the endogenous gene and human type II collagen is found in extracellular matrix surrounding chondrocytes in cartilage. The results indicate that the cis-acting requirements for correct temporal and spatial regulation of the gene are contained within the introduced DNA. PMID- 3033666 TI - Avian sarcoma virus 17 carries the jun oncogene. AB - Biologically active molecular clones of avian sarcoma virus 17 (ASV 17) contain a replication-defective proviral genome of 3.5 kilobases (kb). The genome retains partial gag and env sequences, which flank a cell-derived putative oncogene of 0.93 kb, termed jun. The jun gene lacks preserved coding domains of tyrosine specific protein kinases. It also shows no significant nucleic acid homology with other known oncogenes. The probable transformation-specific protein in ASV 17 transformed cells is a 55-kDa gag-jun fusion product. PMID- 3033665 TI - MYC oncogene involved in a t(8;22) chromosome translocation is not altered in its putative regulatory regions. AB - We have cloned the translocation-associated MYC gene from the Burkitt lymphoma cell line (BL2) with a t(8;22) chromosomal translocation and have determined the nucleotide sequence of the first exon and of the 3' and 5' flanking regions, where sequences with putative regulatory functions have been identified. The nucleotide sequence of the 5' flanking region, which contains regions of DNase hypersensitivity and binding sites for putative regulatory proteins, is the same as that of the normal MYC. Accordingly, mutations in these regulatory regions are not required for the transcriptional deregulation of MYC in the BL2 cell line. The nucleotide sequence of the first exon is similar to that of the normal MYC [Gazin, C., Dupont de Direchin, S., Hampe, A., Masson, J. M., Martin, P., Stehelin, D. & Galibert, F. (1984) EMBO J. 3, 383-387] and has the coding capacity for a 188-residue polypeptide. However, six nucleotide changes that occur in the middle of this reading frame could result in amino acid substitutions. We also have cloned and sequenced the t(8;22) chromosomal breakpoint that is located 10 kilobases 3' of the MYC exon 3 and near the C lambda 3 gene on chromosome 22. Sequences with homology to immunoglobulin joining signals occur close to the breakpoint both on chromosome 8 and 22, providing further evidence that the immunoglobulin joining enzymes may be involved in the recombinations associated with a variety of chromosomal translocations in B and T cells. PMID- 3033667 TI - Isolation of a human major histocompatibility complex class I gene encoding a nonubiquitous molecule expressed on activated lymphocytes. AB - The human major histocompatibility complex is a multigene family containing at least 20 class I genes. Included within this family are the loci encoding the highly polymorphic HLA-A, -B, and -C antigens present at the surface of most nucleated cells. The large number of genes detected with class I probes by Southern blot analysis and the existence of serological reagents defining nonubiquitous, non-HLA-A,B,C class I antigens suggest that products other than HLA-A,B,C antigens are encoded within the class I gene family. These products might be the human counterparts of the murine Qa and TL antigens. In order to identify non-HLA-A,B,C genes, we have developed a probe, JF11, located in noncoding regions flanking the HLA-A locus. This probe detects only a limited number of class I genes and does not detect HLA-A,B,C-associated restriction fragments on Southern blots. This probe was used to screen a human cosmid library. Some of the cosmids isolated with this probe were then transferred into mouse fibroblasts expressing human beta 2-microglobulin. One of the transfectants specifically reacts with one alloantiserum (HA2) that detects HLA class I molecules specific to HLA-A2-positive, phytohemagglutinin-activated T cells and not found on resting T or B cells. Data presented in this paper provide evidence for the isolation and expression of a class I gene encoding a nonubiquitous class I antigen that could be a human analogue of the murine Qa antigens. PMID- 3033668 TI - The same beta-globin gene mutation is present on nine different beta-thalassemia chromosomes in a Sardinian population. AB - The predominant beta-thalassemia in Sardinia is the beta 0 type in which no beta globin chains are synthesized in the homozygous state. We determined the beta thalassemia mutations in this population by the oligonucleotide-probe method and defined the chromosome haplotypes on which the mutation resides. The same beta 39(CAG----TAG) nonsense mutation was found on nine different chromosome haplotypes. Although this mutation may have arisen more than once, the multiple haplotypes could also be generated by crossing over and gene conversion events. These findings underscore the frequency of mutational events in the beta-globin gene region. PMID- 3033669 TI - Intracellular human gamma-interferon triggers an antiviral state in transformed murine L cells. AB - Interaction of human gamma-interferon (IFN-gamma) with a cell-surface receptor is known to be essential for the cell to become resistant to viral infection. Here we demonstrate that IFN-gamma, when present inside the cell, is also capable of inducing a permanent antiviral state. Mouse cells transformed with a truncated human cDNA encoding a mature IFN-gamma protein lacking the signal peptide accumulate high levels of intracellular human IFN-gamma. Not only do these cells acquire a permanent resistance to viral infection, they also exhibit all the biochemical characteristics normally observed after exposure to exogenous IFN. The observed loss of species specificity normally associated with IFN-gamma suggests that this restriction is strictly dependent on the interaction of the molecule with the cell-surface receptor. PMID- 3033670 TI - Allele-specific control of Ia molecule surface expression and conformation: implications for a general model of Ia structure-function relationships. AB - Sequence polymorphism of class II major histocompatibility complex-encoded molecules (Ia) not only accounts for the allelic variability in Ia structure relevant to T-lymphocyte responses but also seems to result in differential quantitative expression of particular Ia heterodimers. The contributions of different allelically variable regions of Ia molecules to both of these processes were analyzed by transfection of L cells with various A beta and A alpha gene pairs. The results show that, with regard to quantitative and qualitative aspects of Ia expression, the polymorphisms in the A beta chain segregate into two groups. Those in the NH2-terminal half of A beta 1 have a consistent role in controlling beta-alpha chain interactions, efficiency of dimer expression, and Ia conformation and probably are in the interior of the Ia molecule at the site of beta-alpha domain interaction. Polymorphisms in the COOH-terminal half of A beta 1 contribute to those structures that directly interact with antibodies, antigen, and/or T-cell receptors, consistent with their presence on the surface of the Ia heterodimer. This analysis provides a model for understanding both overall class II molecular structure and the relationship between this structure and immune recognition. It also suggests an explanation for the evolution of certain features of class II genes. PMID- 3033671 TI - Isolation and characterization of cDNA clones encoding the human carcinoembryonic antigen reveal a highly conserved repeating structure. AB - For the isolation of cDNA clones encoding the carcinoembryonic antigen (CEA), we have constructed a cDNA library from human colon tumor mRNA. The library was screened with various oligonucleotides whose sequence had been deduced from partial amino acid sequence data for CEA. Positive candidate clones were hybridized with a probe for repetitive DNA, because CEA mRNA contains an Alu repetitive element, and with a fragment of a genomic clone of nonspecific cross reacting antigen, an antigen closely related to CEA. Here we report the nucleotide sequence of the two overlapping CEA cDNA clones comprising 1422 nucleotides of CEA mRNA. This sequence encodes the 372 COOH-terminal amino acids of CEA followed by 305 nucleotides of 3' untranslated sequence containing a truncated Alu repeat. The predicted protein sequence is composed of two repeats comprising 178 amino acids, each with an exceptionally high homology of 67%. Each repeat unit contains four conserved cysteine residues and six to nine putative N glycosylation sites. CEA mRNA is most strongly expressed in primary colon tumors and, to a lesser extent, in normal colonic tissue. No CEA mRNA is found in HeLa cells and normal human fibroblasts. PMID- 3033672 TI - Molecular cloning of a gene belonging to the carcinoembryonic antigen gene family and discussion of a domain model. AB - Carcinoembryonic antigen (CEA) is a glycoprotein important as a tumor marker for colonic cancer. Immunological and biochemical studies have shown it to be closely related to a number of other glycoproteins, which together make up a gene family. We have cloned a member of this gene family by using long oligonucleotide probes (42-54 nucleotides) based on our protein sequence data for CEA and NCA (nonspecific cross-reacting antigen) and on human codon usage. The clone obtained (lambda 39.2) hybridizes with six probes and has a 15-kilobase insert. The 5' end of the gene is contained within a 2700-base-pair EcoRI fragment, which hybridizes with five of the six synthetic probes. Sequencing of the 5' end region revealed the location and structure of one exon and two putative intron boundaries. The exon encodes part of the leader sequence and the NH2-terminal 107 amino acids of NCA. Southern blot analysis of human normal and tumor DNA, using as probes two lambda 39.2 fragments that contain coding sequences, suggests the existence of 9 11 genes for the CEA family. One of the restriction fragments described here has been used by Zimmermann et al. [Zimmermann, W., Ortlieb, B., Friedrich, R. & von Kleist, S. (1987) Proc. Natl. Acad. Sci. USA 84, 2960-2964] to isolate partial cDNA clones for CEA. The identity of this clone was verified with our protein sequence data [Paxton, R., Mooser, G., Pande, H., Lee, T.D. & Shively, J.E. (1987) Proc. Natl. Acad. Sci. USA 84, 920-924]. We discuss a domain structure for CEA based on the CEA sequence data and the NCA exon sequence data. It is likely that this gene family evolved from a common ancestor shared with neural cell adhesion molecule and alpha 1 B-glycoprotein and is perhaps a family within the immunoglobulin superfamily. PMID- 3033673 TI - Photodynamic action of merocyanine 540 on artificial and natural cell membranes: involvement of singlet molecular oxygen. AB - The photochemistry of merocyanine 540 (MC 540), a sensitizing dye that binds preferentially to leukemia and electrically excitable cells, has been investigated. MC 540-mediated photooxidation of histidine, arachidonate, and unsaturated phospholipid vesicles was assessed by spin label oximetry and shown to involve type II (singlet oxygen) chemistry. The dye was also shown to be a potent sensitizer of lipid peroxidation in a natural cell membrane, the erythrocyte ghost. Inhibition by azide, stimulation by 2H2O, and identification of the cholesterol product 5 alpha-cholest-6-ene-3 beta,5-diol in this system, all were consistent with singlet oxygen intermediacy. Finally, MC 540 was found to be considerably more phototoxic to K-562 leukemia cells in 2H2O than in H2O. We conclude that singlet oxygen plays a major role in the phototherapeutic effects of this dye. PMID- 3033674 TI - Ubiquitin is detected in neurofibrillary tangles and senile plaque neurites of Alzheimer disease brains. AB - Neurofibrillary tangles (NFT) and neurites associated with senile plaques (SP) in Alzheimer disease-affected brain tissues were specifically immunostained with affinity-purified antibody preparations directed against ubiquitin. In addition, a class of neurites seen in brain regions containing NFT and SP were also specifically stained. Cross-reactivity of the ubiquitin antisera for tau protein, neurofilament proteins, and high molecular weight microtubule-associated proteins (MAPs) were ruled out by (i) the inability of the ubiquitin antisera to stain these proteins in immunoblotting experiments and (ii) the inability of tau, neurofilament, and MAP preparations, when preincubated with the ubiquitin antisera, to inhibit the selective neurofibrillar staining observed. Our results are consistent with the suggestion that ubiquitin is covalently associated with the insoluble neurofibrillary material of NFT and SP. We propose that the ubiquitin-mediated degradative pathway may be ineffective in removing these fibrillar structures in Alzheimer disease brain. PMID- 3033676 TI - Cloning and sequence analysis of cDNAs for neurohypophysial hormones vasotocin and mesotocin for the hypothalamus of toad, Bufo japonicus. AB - The primary structures of the precursors of neurohypophysial hormones vasotocin (VT) and mesotocin (MT) in the hypothalamus of the toad Bufo japonicus were determined by analyzing the nucleotide sequences of the cloned cDNAs encoding them. The MT precursor consists of 125 amino acid residues containing a signal peptide followed directly by MT, which in turn is connected to the MT neurophysin by Gly-Lys-Arg, a processing and carboxyl-terminal amidation signal. In contrast, the VT precursor includes a glycoprotein of 36 amino acids following the VT neurophysin. Except for glycoprotein, the structures of MT and VT precursors are quite similar. RNA transfer blotting analysis showed that both MT and VT mRNAs are present in the brain but not in the liver, ovaries, and testes of the toad. The sequences and the structural organizations of the MT and VT precursors are highly homologous to those of their mammalian counterparts, oxytocin and arginine vasopressin precursors, respectively. This fact suggests that, in the evolutionary pathway of neurohypophysial hormones, VT is the ancestor molecule of vasopressin, while MT is that of oxytocin. PMID- 3033675 TI - Screening an expression library with a ligand probe: isolation and sequence of a cDNA corresponding to a brain calmodulin-binding protein. AB - The use of cloning vectors that express inserted cDNA as fusion protein has led to the isolation of genes encoding a variety of eukaryotic proteins. In these instances antisera or monoclonal antibodies were used as probes to screen expression libraries. Since fusion proteins sometimes display biological activity reflective of the insert-specified portion, we tested the possibility that ligand binding sites might exist in fusion proteins. Specifically we used 125I-labeled calmodulin as a probe to screen a mouse brain lambda gt11 library. One clone, lambda ICM-1 isolated using this approach, produces fusion protein that binds calmodulin with high affinity (Kd, 3-10 nM) in a Ca2+-dependent manner. Molecular genetic mapping experiments and deduction of the predicted higher-order structure from sequence data indicate the binding site is, or is within, a basic, amphiphilic alpha-helical domain composed of approximately 20 amino acids. lambda ICM-1 hybridizes with brain mRNA of 2.1 and 3.5 kb but not with mRNA from liver or kidney, suggesting possible restriction of the protein to brain. We discuss several observations that suggest lambda ICM-1 corresponds to Ca2+/calmodulin dependent protein kinase II, an enzyme that phosphorylates several neuronal proteins, some of which apparently play a role in synaptic function. Our results suggest certain types of ligands may be useful probes to isolate genes encoding various receptor proteins, particularly when the protein is very rare or when it is difficult to obtain antibodies suitable for screening libraries. PMID- 3033678 TI - Free radical mechanisms in relation to tissue injury. PMID- 3033677 TI - Developmentally regulated expression of the nerve growth factor receptor gene in the periphery and brain. AB - Nerve growth factor (NGF) regulates development and maintenance of function of peripheral sympathetic and sensory neurons. A potential role for the trophic factor in brain has been detected only recently. The ability of a cell to respond to NGF is due, in part, to expression of specific receptors on the cell surface. To study tissue-specific expression of the NGF receptor gene, we have used sensitive cRNA probes for detection of NGF receptor mRNA. Our studies indicate that the receptor gene is selectively and specifically expressed in sympathetic (superior cervical) and sensory (dorsal root) ganglia in the periphery, and by the septum-basal forebrain centrally, in the neonatal rat in vivo. Moreover, examination of tissues from neonatal and adult rats reveals a marked reduction in steady-state NGF receptor mRNA levels in sensory ganglia. In contrast, a 2- to 4 fold increase was observed in the basal forebrain and in the sympathetic ganglia over the same time period. Our observations suggest that NGF receptor mRNA expression is developmentally regulated in specific areas of the nervous system in a differential fashion. PMID- 3033680 TI - Brain substrates and the effects of nutrition. PMID- 3033681 TI - Inhibition of cell-associated herpes simplex virus type 2 glycoproteins by delta 9-tetrahydrocannabinol. AB - This study was conducted to define the effect of micromolar concentrations of delta 9-tetrahydrocannabinol (delta 9-THC) on the biosynthesis and expression of herpes simplex virus type 2 (HSV2)-specified glycoproteins. Dose-related reductions in all species of virus glycoproteins were recorded by one-dimensional SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and autoradiography of [14C]glucosamine-labeled infected Vero cells treated with 10(-7) to 10(-5) M delta 9-THC. A drug dose-related depletion of the mature HSV2 major envelope glycoprotein complex (119-kDa average molecular weight), accompanied by accumulation of immature unglycosylated species, was demonstrated by two dimensional SDS-PAGE in concert with Western immunoblotting or autoradiography. Light and electron microscopy immunoperoxidase staining revealed that delta 9-THC effected depletion of 119-kDa determinants from the infected cell surface. This depletion occurred concomitantly with accumulation of 119-kDa components at the perinucleus. However, the expression of 119-kDa glycoproteins on the virion envelope was not affected. These results indicate that delta 9-THC inhibits the synthesis, maturation, and cellular transport of HSV2-specified glycoproteins. Decreased expression of virus glycoproteins on the infected cell surface may affect host immune responsiveness to HSV2. PMID- 3033679 TI - Muscle damage during exercise: possible role of free radicals and protective effect of vitamin E. PMID- 3033682 TI - The inhibition of endotoxin-induced local inflammation by LDH virus or LDH virus infected tumors is mediated by interferon. AB - The footpad swelling reaction induced by local injection of S. marcescens lipopolysaccharide was found to be inhibited in mice given a transplantable tumor (TA3) or cell-free ascitic fluid from tumor-bearing mice. The tumor was shown to contain LDH virus, which is known to cause inapparent persistent infections in mice. Monoclonal antibodies directed against protein VP3 of the LDH virus could partially abrogate the anti-inflammatory effect of the TA3-ascitic fluid, and, conversely, the anti-inflammatory effect could be obtained by LDH virus isolated from the tumor and reproduced by serial passage of cell-free fluids. Inhibition of the footpad reaction was seen in the acute but not in the chronic phase of LDH virus infection, suggesting that the anti-inflammatory effect might be due to endogenous interferon (IFN) which, similarly, was only detectable in the acute phase. Newcastle disease virus, another potent interferon inducer, had a similar inhibitory effect on the footpad reactivity. Moreover, the inhibitory effect of LDH virus infection could partially be abrogated by administration of a polyclonal antibody directed against murine IFN-alpha,beta. Finally, passively administered natural murine IFN-alpha,beta or recombinant murine IFN-alpha 1 (but not recombinant murine IFN-beta) was found to cause inhibition of the footpad reaction. Since Gram-negative bacteria and their lipopolysaccharides have the ability to induce a systemic interferon response, our findings suggest that this interferon may play a modulatory role in local inflammation caused by these bacteria. Our findings also open a new perspective for interferon therapy of certain inflammatory reactions to bacterial infections. PMID- 3033683 TI - Beta adrenergic and muscarinic receptor densities of rat submandibular main duct. AB - [3H]DHA binding studies show that main duct of rat submandibular gland has both beta 1 and beta 2 adrenoceptors, with the percentages of each being 69 and 31%, respectively, whereas whole submandibular gland has 90% beta 1 and 10% beta 2 adrenoceptors. Muscarinic receptors of main duct are 25% less than that of whole submandibular gland. PMID- 3033684 TI - Down-regulation of glucocorticoid and beta-adrenergic receptors on lectin stimulated splenocytes. AB - Activation of porcine splenocytes with the mitogen, concanavalin A, increases the number of glucocorticoid and beta-adrenergic receptors with no change in the apparent dissociation constant. Incubation of splenocytes with concanavalin A in the presence of hydrocortisone 21-sodium succinate prevented this mitogen-induced increase in glucocorticoid receptors. Isoproterenol also prevented the concanavalin A-induced increase in beta-adrenoceptors at 24 hr and reduced the binding affinity of these receptors at 48 hr. Neither agonist had any significant effect on the receptor number of binding affinity of nonstimulated cells. These data demonstrate that the increase in the number of glucocorticoid and beta adrenergic receptors that occur on lymphoid cells after activation by a T-cell mitogen can be prevented by appropriate hormone agonists. Down-regulation of receptor number by appropriate agonists appears to be a common regulatory system that is shared by both the neuroendocrine and the immune systems. PMID- 3033685 TI - Adrenocortical function of the domestic fowl: effects of orchiectomy and androgen replacement. AB - The effect of orchiectomy and androgen replacement on cockerel adrenocortical function was investigated. Orchiectomized cockerels (2 weeks old) were implanted with Silastic tubing containing various amounts of one of the following steroids: cholesterol, testosterone (T), androstenedione (A4), and 5 alpha dihydrotestosterone (DHT). Birds were administered additional implants, containing doses of steroids equivalent to those of the initial implants, at 4 and 8 weeks of treatment (i.e., 6 and 10 weeks of age). Sham-operated cockerels administered empty implants served as intact controls for comparison of data. Animals were killed after 10 weeks of treatment (12 weeks old). Trunk plasma corticosterone (B) and plasma T, and B production by collagenase-isolated adrenocortical cells incubated briefly (2 hr) with or without steroidogenic agents were measured by radioimmunoassay. Orchiectomy with implantation of the inert sterol, cholesterol (hereafter referred to as orchiectomy), did not alter plasma B concentrations and did not affect basal cellular B production or cellular B production induced by a maximal steroidogenic concentration of ACTH or that maximally supported by 25-hydroxycholesterol. However, orchiectomy did lower maximal 8-bromo-cyclic AMP-induced B production by 30%. Low-implant doses of A4 (1-cm implant) and T (0.3-cm implant), that maintained comb growth, lowered plasma B concentrations by 24-42%, whereas a high-implant dose of T (3-cm implant) and all implant doses of DHT had no effect on plasma B concentrations. Thus, androgen replacement had different effects on plasma B depending on the type of androgen and the implant dose. In contrast, androgen replacement consistently suppressed basal and maximal ACTH-induced cellular B production regardless of the type of androgen. Furthermore, the degree of suppression was dose-dependent. These results suggest that the differential effect of androgen replacement on plasma B concentrations was due to differences in the clearance of circulating B and/or differences in blood volume. In addition, the present study suggests that in the absence of the testes, androgens are suppressants of adrenocortical cell function in the domestic fowl. PMID- 3033686 TI - The use of 12-hydroxyheptadecatrienoic acid (HHT) as an HPLC/spectrophotometric marker for cyclooxygenase pathway activity in resident rat peritoneal cells. AB - 12-Hydroxyheptadecatrienoic acid (HHT), a UV chromophore, has been used to assess cyclooxygenase (CO) pathway metabolism of arachidonic acid (AA) by rat peritoneal cells. Simultaneous monitoring at 235 and 280 nm after HPLC permits the measurement of both CO and 5-lipoxygenase (5-LO) pathway fluxes in a single system. PMID- 3033687 TI - The effect of indomethacin on plasma and lung lymph angiotensin converting enzyme activity in sheep. AB - The effect of indomethacin (INDO), an inhibitor of prostaglandin (PG) cyclooxygenase, on pulmonary vascular permeability is unclear. We measured angiotensin converting enzyme (ACE) activity in plasma and lung lymph in order to evaluate pulmonary endothelial integrity. Eighteen sheep, anesthetized and acutely prepared for the collection lung lymph, were used in the study. Four animals (Group I) served as sham controls and were not subjected to experimental intervention. In order to minimize changes in pulmonary microvascular surface area (PMSA), left atrial blood pressure (Pla) was elevated (12-20 Torr) following the baseline period in animals in the three experimental groups. Group II (n = 4) was subjected to increased Pla only in order to assess the effects of that maneuver alone on experimental parameters. Group III (n = 3) received an infusion of buffered vehicle only during a four hour period of elevated Pla, and, thus served as a vehicle control for Group IV (n = 7) which received INDO for the same period of increased Pla. Pulmonary blood pressures, cardiac output (CO), lung lymph flow (Ql) and the ratio of lymph to plasma protein concentration (L/P) were measured in all experiments in order to independently assess changes in lung vascular permeability. While ACE activity in plasma and lymph fell in each experimental group, average ACE experimental values were unchanged from corresponding baseline values. Increased Pla caused a characteristic increase in Ql and fall in L/P in each of the experimental groups. We conclude that INDO does not alter lung fluid and protein balance by a process which involves increased pulmonary vascular permeability secondary to a loss of endothelial integrity. PMID- 3033689 TI - Gastric effects of leukotrienes. AB - The effects of leukotrienes on gastric functions have been outlined by recent experimental and clinical studies. Leukotrienes were found to stimulate pepsin secretion, to reduce mucosal blood flow and to interfere with gastric emptying. Lipoxygenase products appear to impair gastric mucosal integrity and to exacerbate the damaging effects of noxious agents such as absolute ethanol. Drugs acting as leukotriene inhibitors may have a place in the future as anti-ulcer agents. PMID- 3033688 TI - Hemodynamic effects of PAF-acether on the dog kidney. AB - PAF-Acether (PAF) is a potent vasodilator produced in several organs, including the kidney. In the present study, the effect of intra-femoral PAF (0.78 micrograms/kg) in control dogs (Group 1) or indomethacin (3 mg/kg) treated animals (Group 2) was examined. In Group 1, systemic blood pressure dropped from 108 +/- 8 to 47 +/- 7 mmHg following PAF and hematocrit rose from 42 +/- 2 to 56 +/- 2%. These changes were associated with a reduction in both urine flow, urinary sodium (from 69 +/- 9 to 25 +/- 6 muEq/min), glomerular filtration and renal plasma flow (from 28 +/- 2 to 11 +/- 1 and 52 +/- 5 to 26 +/- 4 ml/min, respectively). All parameters returned to normalcy during the following 50 minutes. In Group 2, the systemic effects of PAF were abolished by indomethacin. However, indomethacin failed to prevent the renal abnormalities which were altered as in Group 1. In additional experiments (Group 3) the influence of BN 52021, a specific antagonist of PAF receptors, was examined. The dose of PAF utilized in this group was 0.78 micrograms/kg, whereas BN-52021 was administered 30 minutes before PAF injections in increasing doses (1.0, 2.5, 5.0 and 25.0 micrograms). This antagonist blocked the effect of PAF on blood pressure and renal parameters in a dose-related manner. Finally, the effect of intrarenal PAF was studied (Group 4: increasing continuous infusions of 2, 5, 10, and 20 ng/kg/min; Group 5: single bolus of 0.15 and 0.30 micrograms/kg). In these two groups, the systemic effects of PAF were abolished as expected. In Group 4, a dose related reduction of urinary sodium was observed during the continuous infusion of PAF. Only at higher doses, was an effect on glomerular filtration and renal plasma flow observed. In Group 5, a marked reduction of urinary sodium (from 110 +/- 15 to 22 +/- 5 muEq/min) occurred while glomerular filtration and renal plasma flow decreased by approximately 50%. These data support a direct influence of PAF on urinary sodium excretion and renal hemodynamics. The peripheral effects of this compound are mediated by vasodilatory prostaglandins, as shown in Group 2. Finally, the actions of this powerful vasodilator on the kidney do not require the intervention of systemic influences, as clearly demonstrated in Groups 4 and 5. PMID- 3033690 TI - Specific binding of leukotriene C4 to endothelial cell membranes. AB - Vascular endothelium is a target for leukotriene C4 (LTC4) as demonstrated by previous in vivo and culture experiments. Binding assays were carried out at 0 degrees C on membrane fraction obtained from bovine aortic endothelial cells in culture. Specific binding sites (Kd = 49.9 +/- 6.3 nmol X 1(-1), N = 1.2 X 10(6) sites per cell) for LTC4 were demonstrated in this preparation. Competition studies showed that LTB4, LTD4 and LTE4 did not displace LTC4 from its binding sites. FPL 55712, a sulfidopeptide antagonist, was seen to be a weak competitor and reduced glutathione exhibited a significant affinity for the binding site. The possible receptor role of this site is discussed. PMID- 3033691 TI - The influence of dietary marine oil (Polepa) and evening primrose oil (Efamol) on prostaglandin production by the rat mesenteric vasculature. AB - The interactions of n-6 and n-3 fatty acids on prostaglandin metabolism in the isolated rat mesenteric vessels were studied. Sprague-Dawley rats (200-220 g) were fed for two weeks a fat-free semi-synthetic diet supplemented with 10% by weight of different combinations of Evening Primrose Oil (Efamol), a rich source of linoleic acid (LA) and gamma-linolenic acid (GLA), the immediate precursor of dihomo-gamma-linolenic acid (DGLA), and Polepa (POL), a marine oil rich in eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids. The combinations of supplement were as follows: 9% Efamol - 1% POL, 8% Efamol - 2% POL, 7% Efamol - 3% POL, 6% Efamol - 4% POL, 5% Efamol - 5% POL. The outflow of thromboxane (TxB2), prostacyclin (6-keto-PGF1 alpha), PGE2, and PGE1 was decreased in relation to the proportion of marine oil in the diet, except for the group which received 8% Efamol - 2% POL, and which showed an increase in 6-keto-PGF1 alpha, PGE2, and PGE1. The decrease in TxB2 was much greater than those of 6-keto-PGF1 alpha or PGE2, while PGE1 followed the same pattern as prostacyclin and PGE2. These results suggest that n-3 fatty acids, at high concentrations, inhibits conversion of both DGLA and AA to eicosanoids. Low concentrations of fish oil may, in contrast, increase formation of desirable 1 and 2 series eicosanoids. PMID- 3033692 TI - Effects of chronic ethanol ingestion on arachidonic acid metabolism in rat tissues and in vitro effect of ethanol on cAMP in platelets. AB - Effects of chronic ethanol ingestion (3 weeks) on the capacity of peritoneal macrophages, lung and heart tissues, to metabolize endogenous arachidonic acid (AA) in rats, and the in vitro effect of ethanol on 3'-5' cyclic adenosine monophosphate (cAMP) levels in rat platelets were studied. Peritoneal resident macrophages were stimulated by calcium ionophore (A23187) and levels of 5 hydroxyeicosatetraenoic acid (5-HETE), leukotriene B4 (LTB4) and PGE2 were measured by radioimmunoassay. There were no differences in levels of the eicosanoids synthesized by macrophages between the ethanol treated and the control group. There were also no differences found in levels of the eicosanoids synthesized by heart or lung homogenate between the two groups. These results suggest that chronic ethanol ingestion does not alter the capacity to synthesize the eicosanoids from the endogenous precursor in tissues studied here. Preincubation of ethanol with platelet rich plasma resulted in a dose dependent increase in cAMP levels. The well documented inhibitory effects of ethanol in vitro on aggregation and AA metabolism in platelets may be due to the enhanced cAMP levels. PMID- 3033694 TI - Mechanism of lithium action: in vivo and in vitro effects of alkali metals on brain superoxide dismutase. AB - Intraperitoneal administration of lithium (2 mEq/kg/day) was found to increase the superoxide dismutase (SOD) activity in certain brain regions after 24 hours (2 injections) and 3 days (once a day) of exposure. In vitro addition of wide range of lithium (0.1 to 8 mEq) to enzyme preparation as well activated cortical SOD activity; however, at 10 mEq concentrations an inhibition was observed. The increase in SOD activity did not appear to be region specific as under both in vivo and in vitro conditions lithium enhanced enzyme activity in all the tested brain regions. The effects of intraperitoneal administration of 2 mEq/kg rubidium and cesium for 24 hr (2 injections) and 6 days (once a day) were also studied on central SOD. Both the alkali metals were not found to produce any significant alteration in the cortical enzymic activity. When the in vitro effects of these monovalent alkali metals were tested, only 2 mEq rubidium was found to increase cortical SOD; however, cesium and potassium at similar concentration did not produce any appreciable effects. It appears from the data that lithium-induced increase in brain SOD activity is not an unspecific effect of alkali metals. SOD enzyme disposes cytotoxic superoxide radicals which, if not removed, could impair the normal functioning of cellular membrane and produce a variety of psychedelic compounds as well. The activation of central SOD by lithium would enhance the disposal process of superoxide radicals whose pathological concentrations may be present in affective disorders. The mechanism of lithium-induced activation of SOD, at present, is not known. PMID- 3033693 TI - The effect of chronic imipramine and electroconvulsive shock treatment on [3H]DADLE binding to cortical membranes of rats pretreated with chronic reserpine or 6-hydroxydopamine. AB - Repetitive electroconvulsive shock treatment (for 8 days) or chronic administration of imipramine (10 mg/kg/day IP for 14 or 21 days) elevated the density of opioid delta receptors in the cerebral cortex of the rat. Electroconvulsive shock treatment produced a similar effect in rats treated subchronically with reserpine or receiving intraventricularly 6-hydroxydopamine, but these manipulations of central catecholamines prevented the action of imipramine. PMID- 3033695 TI - Differential effects of benzodiazepine receptor ligands on isotonic saline and water consumption in water-deprived rats. AB - Water-deprived male rats were adapted to a 30 min test of water or saline drinking in a single-bottle acceptance test. The potent benzodiazepine agonist, clonazepam, produced significant increases in both water and saline consumption. Increases in the consumption of both were also obtained with the non benzodiazepine agonist, zopiclone (a cyclopyrrolone), but not with the pyrazoloquinoline agonist, CGS 9896. Hence, some, but not all, benzodiazepine receptor agonists enhance drinking responses. The benzodiazepine receptor antagonists, Ro15-1788 and CGS 8216, had no significant effect on the intake of either isotonic saline or water. In contrast, the beta-carboline FG 7142, which has been described as an inverse agonist acting at benzodiazepine receptors, reduced both saline and water drinking at 10 and 20 mg/kg. Although the baseline level of saline drinking was considerably higher than that of water, there was no general indication that any drug effect on consumption interacted with the type of fluid in the drinking test. However, in the case of agonist-induced increases in consumption, peak effects occurred at different doses; they were lower for saline- than for water-drinking. PMID- 3033696 TI - Effect of chronic nicotine treatment against repeated immobilization stress. AB - Alpha 2 and beta adrenoceptors, and muscarinic cholinoceptors in 2 brain regions (cerebral cortex and hippocampus) were measured in rats which received either tap water or nicotine added to the drinking water (5-8 mg/kg/day) for 4 weeks, and immobilization stress (daily 2 hr) for the last 5 days. The repeated stress induced a reduction in the maximum number of binding sites (Bmax) for (3H)dihydroalprenolol (DHA) in the cerebral cortex of rats with tap water, without affecting (3H)clonidine binding. Nicotine-treatment also caused a decrease in the Bmax of cortical (3H)DHA binding comparable to the case of stress, and increased the (3H)clonidine binding. However, the combination of nicotine- and stress-treatments failed to induce further no changes in the 2 radioligands binding. The binding of (3H)quinuclidinyl benzilate in the cerebral cortex and of the 3 radioligands in the hippocampus was unaltered by nicotine- and/or stress-treatments. These results indicate that long-term administration of nicotine induces down-regulation of cortical beta adrenoceptors and seemingly attenuates the receptor alteration by repeated stress. PMID- 3033697 TI - Hypophysectomy prevents ACTH-induced yawning and penile erection in rats. AB - The intracerebroventricular administration of ACTH1-24 (3-5 micrograms/rat) produced a behavioural syndrome characterized by recurrent episodes of penile erection and yawning in rats. Hypophysectomy prevented ACTH1-24-induced yawning and penile erection. These results suggest that pituitary has a "trophic" action not only on peripheral target organs but also on structures in brain controlling specific behavioural responses. PMID- 3033698 TI - Hypothermia: role of alpha 1- and alpha 2-noradrenergic receptors in the hypothalamus of the cat. AB - The purpose of this study was to characterize the alpha 1- and alpha 2 noradrenergic receptor sub-types which could mediate the hypothermic response produced by norepinephrine (NE) and other alpha-noradrenergic agonists applied to the thermosensitive zone of the hypothalamus. An array of four guide tubes was implanted stereotaxically so that their tips rested just above the anterior hypothalamic, preoptic area (AH/POA) of the cat. Following post-operative recovery, a micro-injection of an agonist or antagonist of NE receptors or control CSF vehicle was given in a volume of 1.0-2.0 microliter in the AH/POA in each of the unrestrained cats. The alpha 1-noradrenergic receptor agonist, phenylephrine, but not methoxamine, applied to the AH/POA produced a dose dependent hypothermia of up to 2.0 degrees C. When applied similarly, the alpha 2 noradrenergic agonist clonidine, as well as norepinephrine, which acts on both alpha 1- and alpha 2-noradrenergic receptors, also induced a decline in the cat's core temperature of up to 1.5 degrees C. The hypothermic response of clonidine was inhibited by pre-treatment of the AH/POA with a micro-injection of the selective alpha 2-noradrenergic blocking agent, yohimbine. However, yohimbine given similarly in the cat's AH/POA potentiated significantly both the phenylephrine and norepinephrine-induced hypothermia. The combined alpha 1-, alpha 2-noradrenergic receptor antagonist, phentolamine, also injected into AH/POA inhibited the thermolytic response evoked by both phenylephrine and norepinephrine, whereas it was virtually ineffective against the clonidine induced hypothermia. These results, therefore, strongly suggest that both alpha 1 and alpha 2-noradrenergic receptors subserve the coordinated thermoregulatory mechanisms in AH/POA which are required for the functional dissipation of body heat and the consequent evocation of hypothermia. PMID- 3033699 TI - Differential biochemical mechanisms mediate locomotor stimulation effects by caffeine and nicotine in rats. AB - Effects of caffeine and the interactive effects of caffeine and nicotine on locomotor activity in rats were examined in the present study. Other than confirming previous reports that both drugs enhanced locomotion, we have also found that their effects on activity were additive. Meanwhile, results of various biochemical measures have revealed that at the minimum effective doses of caffeine and nicotine which facilitated locomotor activity, only one biochemical system was preferentially influenced by either drug alone. The most significant findings were that caffeine stimulated the release of catecholamines and nicotine decreased the concentrations of tyrosine and tryptophan in brain. The combined effects of caffeine and nicotine on these brain amines were not different from those of each drug alone. Together with the report that caffeine and nicotine had differential actions on different activity measures, the present results support the hypothesis that caffeine and nicotine affect locomotor activity via different neurochemical mechanisms. PMID- 3033700 TI - Cocaine modulation of central monoaminergic neurotransmission. AB - Extracellular microelectrode studies were conducted to test the effects of cocaine HCl on the activity of spontaneously firing single serotonergic dorsal raphe (DRN), noradrenergic locus coeruleus (LC) and dopaminergic ventral tegmental (VTA) and zona compacta (ZC) neurons, and cerebellar Purkinje neurons (PC) in urethane anesthetized rats in vivo. Cocaine (0.0625-4 mg/kg) predominantly inhibited all of the central monoaminergic neurons and predominantly activated cerebellar Purkinje neurons. Cocaine (1 mg/kg, IV) failed to potentiate the inhibitory effects of LC stimulation on PC neurons. The temporal effects of intravenous cocaine on arterial pressure (i.e., pressor response) were not directly correlated with the effects on neurons. Cocaine did not decrease the amplitude or slope of neuron action potentials, and the effects of cocaine on firing rate were not shared by similar doses of procaine. Reserpine pretreatment (10 mg/kg, IP) attenuated the effects of cocaine (1 mg/kg, IV) on DRN, LC, and PC neurons. Specific adrenoceptor antagonists antagonized the inhibitory effects of cocaine on LC (piperoxane, yohimbine) and VTA (haloperidol) neurons. These results suggest that the central effects of cocaine on presynaptic monoaminergic neurons may in part be mediated by augmented monoamine neurotransmission at autoreceptors and that the effects of cocaine on postsynaptic target cells (PC) may be more complex, requiring the analysis of both pre- and postsynaptic elements. PMID- 3033701 TI - GABAergic mechanisms of analgesia: an update. AB - Both directly acting (GABAA and GABAB agonists) and indirectly acting GABAergic agents (GABA uptake inhibitors and GABA-transaminase inhibitors) produce analgesia in a variety of animal test systems. Analgesia produced by GABAA agonists is probably due to a supraspinal action, although spinal sites may also play a role. GABAA agonist analgesia is insensitive to naloxone, bicuculline, picrotoxin and haloperidol, but is blocked by atropine, scopolamine and yohimbine suggesting a critical role for central cholinergic and noradrenergic pathways in this action. The lack of blockade by the GABAA antagonist bicuculline is difficult to explain. Both bicuculline and picrotoxin have intrinsic analgesia actions which may not necessarily be mediated by GABA receptors. The GABAB agonist baclofen produces analgesia by actions at both spinal and supraspinal sites. Baclofen analgesia is insensitive to naloxone, bicuculline and picrotoxin, and blockade by cholinergic antagonists occurs only under limited conditions. Catecholamines are important mediators of baclofen analgesia because analgesia is potentiated by reserpine, alpha-methyl-p-tyrosine, phentolamine, ergotamine, haloperidol and chlorpromazine. A role for serotonergic mechanisms is less well defined. Methylxanthines, which produce a clonidine-sensitive increase in noradrenaline (NA) turnover, increase baclofen analgesia by a clonidine-sensitive mechanism. Both ascending and descending NA pathways are implicated in the action of baclofen because dorsal bundle lesions, intrathecal 6-hydroxydopamine and medullary A1 lesions markedly decrease baclofen analgesia. However, simultaneous depletion of NA in ascending and descending pathways by locus coeruleus lesions potentiates baclofen analgesia suggesting a functionally important interaction between the two aspects. Baclofen analgesia within the spinal cord may be mediated by a distinct baclofen receptor because GABA does not mimic the effect of baclofen and the rank order of potency both of close structural analogs of baclofen as well as antagonists differs for analgesia and GABAB systems. The spinal mechanism may involve an interaction with substance P (SP) because SP blocks baclofen analgesia, and desensitization to SP alters the spinal analgesic effect of baclofen. GABA uptake inhibitors produce analgesia which is similar to that produced by GABAA agonists because it is blocked by atropine, scopolamine and yohimbine. Analgesia produced by GABA-transaminase inhibitors is similar to that produced by GABAA agonists because it can be blocked by atropine, but it is potentiated by haloperidol while THIP analgesia is not.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3033702 TI - Monosodium glutamate does not alter ACTH- or apomorphine-induced penile erection and yawning. AB - The effect of the intracerebroventricular (ICV) injection of ACTH 1-24 (1, 5 and 10 micrograms) or the subcutaneous administration of apomorphine (20 and 80 micrograms/kg SC) on spontaneous penile erection and yawning was studied in rats treated with monosodium glutamate (MSG), a treatment that depletes hypothalamic ACTH, alpha-MSH and endorphin-like peptides. Neonatal MSG treatment failed to antagonize either apomorphine- or ACTH-induced yawning in male and female rats, or to alter the number of penile erection episodes induced by the two substances in male rats. In contrast, hypophysectomy, that does not alter the concentration of hypothalamic ACTH and alpha-MSH, caused a marked prevention of apomorphine- and ACTH-induced responses, in agreement with previous studies. The results suggest that the integrity of opiomelanotropinergic neurons in the hypothalamus is not necessary for the induction of yawning and penile erection by ACTH-derived peptides, and that apomorphine and other dopamine agonists apparently do not induce penile erection and yawning by releasing an ACTH-derived peptide in brain. PMID- 3033703 TI - Acute delta 9-tetrahydrocannabinol exposure: effects on hypothalamic-pituitary testicular activity in mice. AB - The endocrine functions of the median eminence, pituitary and testes were examined in male mice after exposure to delta 9-tetrahydrocannabinol (THC) either in vivo or in vitro. The secretion of luteinizing hormone-releasing hormone (LHRH) under basal conditions, and in the presence of norepinephrine (NE; 60 microM), was significantly enhanced in median eminence fragments obtained 1 hr post-treatment with THC (50 mg/kg), while addition of THC (250 ng/ml) to the incubation media enhanced clonidine, as well as NE-stimulated LHRH release, but did not affect basal LHRH release. In vitro exposure to THC also enhanced LHRH stimulated LH release by pituitaries, but did not affect basal secretion rates. In vivo THC exposure tended to enhance pituitary responsiveness to LHRH, although this effect was not statistically significant. In testicular perifusions, addition of THC at a concentration of 250 ng/ml completely blocked hCG-stimulated T secretion within 30 min. The suppressive effects of a lower dose of THC, 25 ng/ml, required 60 min to inhibit T production, an effect which persisted for 60 80 min post-THC. These findings indicate that THC exposure enhances responsivity at neuroendocrine target sites, but attenuates gonadotropin-stimulated testicular steroidogenesis. PMID- 3033704 TI - Atropine sulfate increases pituitary responses to stress. AB - The effects of atropine sulfate pretreatment on pituitary indices of stress response were examined. Pituitary cyclic AMP and plasma prolactin increases following 15 min of acute stress were used as measures of stress response. Over a range of doses (0, 5, 10, 30 and 60 mg/kg), pretreatment with atropine sulfate increased the measured stress responses to footshock but had little or no effect on resting or non-stressed levels of the substances measured. The effects of atropine on response to immobilization were tested only at 5 mg/kg. At this dose, atropine sulfate, but not methylatropine nitrate, increased pituitary cyclic AMP response to immobilization stress demonstrating that the potentiation of the pituitary cyclic AMP stress response was not limited to footshock stress and suggesting that this effect of atropine was central rather than peripheral. Neither atropine nor methylatropine pretreatment at this dose potentiated prolactin response to immobilization stress. PMID- 3033705 TI - The effect of chlordiazepoxide on the habituation of exploration: interactions with the benzodiazepine antagonist RO 15-1788. AB - Rats tested on two occasions in a holeboard apparatus showed between-session habituation of exploratory activity. No habituation was observed on the measure of locomotor activity. Administration of chlordiazepoxide before the first test reduced exploratory behavior in this test and also reduced the degree of between session habituation. Administration of RO 15-1788 reversed the effect of chlordiazepoxide on exploration in the first test, but failed to reverse the drug's effect on between-session habituation. It is unlikely that state-dependent retrieval could account for these results. The results are discussed in relation to the effects of benzodiazepines on learning and memory. PMID- 3033706 TI - Studies of thyrotropin-releasing hormone (TRH)-induced defecation in cats. AB - In unanesthetized cats, defecation produced by thyrotropin-releasing hormone (TRH) was investigated after its injection into the cerebral ventricle (ICV) through chronically implanted cannulae. TRH injected in doses from 0.1 to 1.0 mg into the cerebral ventricle evoked defecation which was not dose-dependent. The antimuscarinic drug, atropine, the ganglionic blocker, mecamylamine, the alpha and beta adrenergic blocking agents, yohimbine and propranolol, the dopamine antagonist, chlorpromazine, the 5-hydroxytryptamine antagonist, methysergide, and the antihistamine, antazoline, all injected into the cerebral ventricle had virtually no effect on the defecation evoked by TRH injected similarly. In cats pretreated with ICV reserpine, 5,6-dihydroxytryptamine and hemicholinium-3, the defecation induced by ICV TRH was not significantly changed. On the other hand, in cats pretreated with ICV 6-hydroxydopamine, the defecation caused by ICV TRH was potentiated. Therefore, it is concluded that TRH-induced defecation could not be related to central catecholaminergic, 5-hydroxytryptaminergic and cholinergic receptors, but rather to central TRH sites in the cat. PMID- 3033707 TI - Interaction of 2,3-benzodiazepines with peripheral benzodiazepine receptors. AB - 2,3-Benzodiazepines (BZs), such as tofizopam (TP) and GYKI-51 189 have anxiolytic potency accompanied by moderate sedative action, but no anticonvulsant and muscle relaxant activities. These compounds show relatively low affinity to the peripheral benzodiazepine (PBZ) receptors, nevertheless, they decrease the binding of (3H)Ro5-4864 to its receptors in heart, kidney and brain membranes. This diminution in the binding is due to a decrease in the affinity for the ligand (Kd) without any change in the maximal number of binding sites (Bmax). This interaction of 2,3-BZs with PBZ binding sites may explain their pharmacological profile. PMID- 3033708 TI - Catecholamine action on smooth muscle. PMID- 3033709 TI - Responses of renal, coronary and vertebral vasculatures to MC-838 and captopril in anesthetized dogs. AB - The regional hemodynamic effects of MC-838, a new angiotensin-converting enzyme inhibitor, and captopril at equidepressor doses were examined in the anesthetized dog by simultaneously measuring renal (RBF), coronary (CBF), vertebral (VBF) arterial and aortic blood flow (AoF). Hemodynamic responses to angiotensin I (AI), AII and noradrenaline were compared before and after the administration of each inhibitor. MC-838 (3 mg/kg i.v.) lowered gradually aortic pressure (AoP) and increased moderately AoF and RBF up to 60 min after the administration. Captopril (0.1 mg/kg i.v.) lowered AoP immediately after the administration and increased AoF and RBF more shortly than MC-838. Neither inhibitor produced a marked change in VBF or CBF. The effects of the inhibitors in the renal vascular bed was much greater than that in vertebral and coronary vascular beds, although vascular resistance in all of them was significantly reduced. Each of the drugs inhibited the pressor and renal vasoconstrictor responses to AI. These results indicate that the renal vasculature is more sensitive to both MC-838 and captopril than vertebral and coronary vasculature, but MC-838 has a slower and longer-lasting action than does captopril. PMID- 3033710 TI - Stopping power and ranges of low-energy protons in tissue-equivalent gas. AB - Stopping power data for protons in tissue-equivalent gas (64.4% CH4 + 32.4% CO2 + 3.2% N2, partial pressures) were derived from experimentally determined ranges and from differential ionisation distributions using the differential energy required per ion pair formed to convert ionisation to energy loss. The proton energy covered the region from 1 to 100 keV. The results are compared with other data available. In general the present experimental stopping power data tend to higher values than calculated data for energies lower than 20 keV; between 20 and 100 keV they agree well within the stated uncertainties. PMID- 3033711 TI - [Medical treatment of the Raynaud phenomenon]. AB - The medical treatment of Raynaud's phenomenon must be adapted to each particular case. Benign cases do not require drug therapy. Various substances seem able to relieve the most severe cases (calcium blockers, alpha-blockers). Prostaglandins perfusions and plasmaphoresis must be reserved for the most severe cases, with trophic disorders not improved with less aggressive treatments. PMID- 3033712 TI - [Ectopic form of the Klippel-Trenaunay syndrome]. PMID- 3033713 TI - 2,7-Diiodo-9-diazofluorene: a potential reagent for photolabeling. PMID- 3033714 TI - Effect of humidity on photoinduced ESR signal from human hair. PMID- 3033715 TI - Synergism of a compound unconditioned stimulus in taste aversion conditioning. AB - Anti-lymphocyte serum (ALS) and lithium chloride have both previously been used successfully as unconditioned stimuli in taste aversion conditioning paradigms in rats. This report substantiates those findings but shows that when the two stimuli are given as a compound unconditioned stimulus in association with saccharin flavoured drinking solution, the conditioned taste aversion response following a second exposure to saccharin alone is more profound than that following conditioning with either ALS or LiCl alone. These results demonstrate synergism between the two stimuli when given together. PMID- 3033716 TI - Naltrexone reverses a long term depressive effect of a toxic lithium injection on saccharin preference. AB - In 2 experiments, using female rats, we demonstrated that an injection of 2% body weight 150 mM lithium chloride (LiCl) reduced saccharin preference, measured 48 hr later. Furthermore, Experiment 1 showed that this effect of lithium was reversed by 10 mg/kg naltrexone injected immediately before lithium administration. Experiment 2 demonstrated that one injection of 10 mg/kg morphine also depressed saccharin preference 48 hr post-injection. Paradoxically, morphine given before lithium reversed the effects of lithium on saccharin preference. The experiments suggest the involvement of endogenous opioid peptides in the long term toxic effects of lithium on saccharin preference. PMID- 3033717 TI - Physical characterization of Rhizobium meliloti megaplasmids. AB - Intact megaplasmids of Rhizobium meliloti 2011 have been isolated and visualized by electron microscopy. The contour lengths of 64 megaplasmid molecules were determined. One definite class of molecules of 400 micron length and a range of larger molecules with lengths of up to 560 micron was observed. The contour lengths of the megaplasmids pRme2011a and pRme2011b were measured after isolation from plasmid-free Agrobacterium strains into which they had been individually transferred. Plasmid pRme2011a corresponds to the 400-micron class of megaplasmids while plasmid pRme2011b belongs to the 560-micron class. Preparatively isolated megaplasmids pRme2011a and b showed completely different restriction patterns. The pattern of total megaplasmid DNA from R. meliloti 2011 is composed of those from pRme2011a and b, suggesting that no more than two different megaplasmids exist. Because the length distributions of measured molecules were broad, R. meliloti 2011 megaplasmids seem to vary in length in vivo. Because only pRme2011a hybridized with a nifHD probe, this is the Sym plasmid. For R. meliloti strain MVII-1, which carries the megaplasmids pRmeMVII 1f and pRmeMVII-1g, pRmeMVII-1f was shown to be the Sym plasmid. Buoyant density determinations of R. meliloti 2011 and MVII-1 megaplasmids gave a value of 1.717 g/cm3 for pSym, which is that of Agrobacterium DNA. The buoyant density of the second megaplasmid was 1.721 g/cm3, corresponding to the density of the R. meliloti chromosome. As determined by reassociation kinetics, pRme2011a and b are unrelated. The degree of relatedness between strains MVII-1 and 2011 was 82%. PMID- 3033718 TI - Characterization of the drug resistance plasmid NTP16. AB - A functional and physical analysis of the multicopy plasmid NTP16 is presented. The plasmid-encoded drug resistance determinants are located, as are regions encoding the origin of replication, incompatibility functions, copy number determinants, and mobility functions. It is demonstrated that NTP16 probably arose from the closely related plasmid NTP1 by the acquisition of a novel kanamycin resistance transposon, Tn4352, followed by deletion of some NTP1 sequences. The incompatibility behavior of NTP16 derivatives indicates a system of control rather more complex than that which operates in ColE1. In addition to the RNA I/primer RNA system, the production of a further trans-acting product is demonstrated and its site of action located. A series of derivative plasmids have been created which may prove useful as vectors for genetic engineering. PMID- 3033719 TI - A pathway for the evolution of the plasmid NTP16 involving the novel kanamycin resistance transposon Tn4352. AB - The kanamycin resistance determinant of the drug resistance plasmid NTP16 has been characterized by DNA sequencing and has been shown to possess all of the structural features of a transposable element. It is made up of a 1040-bp central region encoding a protein identical to the aminoglycoside 3'-phosphotransferase of Tn903, flanked by direct repeats of an element identical to IS26. This novel transposon has been designated Tn4352. Analysis of the host sequences flanking the transposon reveal that they are derived from a Tn3-like element, and contain no 8 base pair target size duplications which are normally created by the insertion of IS26-like elements. Comparison to the Tn3 sequence shows that the flanking sequences are noncontiguous within Tn3, with the clear implication that NTP16 has evolved from a similar plasmid encoding only ampicillin resistance (presumably NTP1) by the insertion of Tn4352 into the Tn3-like element, followed by a substantial deletion. The sequence analysis suggests that the initial insertion was into the tnpR gene of the ampicillin transposon, followed by a deletion extending to a specific site within tnpA. PMID- 3033721 TI - Interactions between the transposable element IS21 on R68.45 and TN7 in Pseudomonas aeruginosa PAO. AB - Tn7 transposes from the chromosome of Pseudomonas aeruginosa into the plasmid R68.45 with tandem IS21, at up to 400 times the frequency that it transposes into R68, which has only one copy of IS21. While R68::TN7 derivatives are stable, R68.45::Tn7 isolates undergo frequent deletions. Instability of R68.45 occurs whether Tn7 is inserted into the plasmid (cis configuration) or into the bacterial chromosome (trans configuration). The deletions of R68.45 start at the junction between the tandem IS21 copies and proceed clockwise, ending in the region of oriT. It appears that Tn7 and IS21 can mutually stimulate transposition of each other. PMID- 3033720 TI - Physical analysis of the conjugative shuttle transposon Tn1545. AB - The conjugative shuttle transposon Tn1545 from Streptococcus pneumoniae confers resistance to kanamycin (aphA-3), erythromycin (ermAM), and tetracycline (tetM). The 25.3-kb element is self-transferable to various gram-positive bacterial genera where it transposes. Tn1545 is also capable of transposition, but not of conjugation, after cloningoff Escherichia coli. Analysis of the element by restriction endonucleases, molecular cloning, electron microscopy of heteroduplexes, DNA hybridization, and sequencing allowed us to establish a physical map of Tn1545, localize the resistance genes, determine their direction of transcription, and compare them with other characterized resistance determinants, and show that Tn1545 is not flanked by large terminal repeated sequences in opposite orientation. PMID- 3033722 TI - Sequence and functional analysis of a divergent promoter from a cryptic plasmid of Lactobacillus acidophilus 168 S. AB - We have characterized three of at least five plasmids borne by Lactobacillus acidophilus 168 S. Restriction mapping indicates extensive sequence homology between at least two of them (p1 and p3). We have cloned them in Escherichia coli, and for the smallest (p1) we present the sequence of a region with two divergently arranged promoters which probably share a symmetrical (TTTAAA)-35 box and function efficiently in E. coli cells; an open reading frame contiguous to the promoter, which codes for a 120 amino acid protein of unknown function, and is transcribed in E. coli; and a transcription termination sequence next to this open reading frame. The promoter region contains an AT cluster which is similar to that of the ori2 region of the E. coli F plasmid, and is probably involved in the control of the replication of p1. PMID- 3033723 TI - Correction. A revision of the nucleotide sequence and functional map of pUB110. PMID- 3033724 TI - Inhibition of angiotensin I converting enzyme by flavanolic compounds: in vitro and in vivo studies. PMID- 3033725 TI - Utilization of glutamate accumulating bacterial cells for production of ribonucleotides. AB - Corynebacterium glutamicum cells are industrially used for glutamate production. However, the waste that contains microbial cells, cellular debris, residual sugars, ammonia and metabolites seriously pollutes the environment. The cells are recovered and utilized for ribonucleotide production so that the pollution caused by the cells is eliminated. Nucleic acid is extracted from the cells and is hydrolyzed with nuclease P1 from Penicillium citrinum. The hydrolysate is fractionated with Dowex-50 and Dowex-1 into 5'-CMP, 5'-UMP, 5'-GMP and 5'-AMP. The products are characterized by electrophoresis, ultraviolet light absorbance, and 5'-nucleotidase. PMID- 3033726 TI - Alpha 2-adrenergic receptors in platelet membranes of depressed patients: increased affinity for 3H-yohimbine. AB - Specific binding to alpha 2-adrenergic receptors was studied in the platelets of 31 patients with major depressive disorder and 18 normal controls using the selective antagonist 3H-yohimbine. Receptor density for depressed patients (Bmax = 88 +/- SD 45.1 fmoles/mg) was not significantly lower than that for controls (124 +/- SD 78.1 fmoles/mg). The affinity of the receptor for yohimbine was significantly greater in depressed patients (Kd = 1.05 +/- SD 0.47 nM) than in controls (Kd = 1.47 +/- SD 0.63 nM). This is consistent with the hypothesis of increased alpha 2-adrenergic receptor sensitivity in depressive disorders. Past studies of alpha 2-adrenergic receptors on platelets are reviewed, and the importance of designing studies with sufficient statistical power is discussed. PMID- 3033727 TI - The interrelationship of beta endorphin, ACTH and cortisol in depressive illness: a controlled study. AB - Plasma cortisol, ACTH and beta endorphin were measured before and after dexamethasone in 8 severely depressed patients and 8 age- and sex-matched controls to examine the relationship of ACTH and endogenous opioids to cortisol in depression. Despite having significantly higher plasma levels of cortisol than the controls, the depressed patients did not have correspondingly elevated plasma levels of ACTH. Beta-endorphin levels were also similar in the two groups. All three hormones suppressed to some degree after dexamethasone, but cortisol suppressed less in patients than controls. Our findings suggest that in severe depressive illness abnormalities exist in the hypothalamic-pituitary-adrenal axis peripherally as well as centrally. PMID- 3033728 TI - Biological aspects of depression. AB - Several neuroendocrine studies with the clonidine-growth hormone stimulation test show endogenous depressive patients to have a reduced GH response as compared to neurotic depressives, schizophrenics and controls. The blunted GH response to clonidine seems to be a trait marker or vulnerability factor for mono- and bipolar endogenous depression. It could be explained by a reduced postsynaptic alpha 2-adrenoceptor sensitivity or of structures related to them. The sensitivity of alpha 2-adrenoceptors is influenced by both, the endorphinergic and cholinergic systems. First results supporting this concept are presented and discussed. PMID- 3033729 TI - New pharmacological findings in depression. AB - The last decade of psychopharmacological research in depression is characterised by the development of structurally novel and biochemically selective-acting drugs and by the shift of interest from the acute to long-term treatment effects of antidepressants on catecholaminergic transmission. Of particular importance in this respect are the findings demonstrating adaptative changes occurring at receptor levels which have led to alternative theoretical formulations about the origin of depression and the mode of action of antidepressants. However in spite of the progress in this field of research the mechanism of therapeutic action of antidepressants remains unknown. Examples of drugs not affecting NA and 5-HT uptake or release and producing no adaptative changes of adrenergic receptors (trimipramine, levoxaprotiline) indicate that antidepressant-like activity can be achieved by mechanisms which still remain to be elucidated. PMID- 3033730 TI - Pleomorphic adenoma of a minor salivary gland: report of a case. PMID- 3033731 TI - Radiation damage to histone H2A by the primary aqueous radicals. AB - Histone H2A has been examined for radiation-induced changes in structure and in amino acid composition. The effects of the individual radical intermediates--the hydroxyl radical, solvated electron, and superoxide radical--have been determined by irradiating in dilute aqueous solution under controlled environmental conditions. Amino acid analysis of irradiated histone H2A shows a selective attack on a few residues; only the aromatic residues, phenylalanine and tyrosine, and the heterocyclic residue, histidine, are significantly decreased. A significant increase in aspartic acid is also observed. Sodium dodecyl sulfate polyacrylamide gel electrophoresis shows that the hydroxyl radical is the effective radical for promoting changes in protein structure. PMID- 3033732 TI - [Structural organization of chromatin as a factor determining the rate of its endonucleolysis in irradiated and nonirradiated thymocytes]. AB - A study was made of chromatin endonucleolysis in hypotonic thymocytes incubated in digestive buffers containing different concentrations of potassium, magnesium, calcium, and mercaptoethanol. Inhibition of endonucleolysis by univalent cation during the first 20 min of incubation was followed by intensive chromatin degradation. A decrease in free potassium content retarded chromatin degradation and enhanced the inhibiting effect of the univalent cations. The regularities of changes in the rate of chromatin endonucleolysis in different digestive buffers were similar with both exposed and intact thymocytes. PMID- 3033733 TI - [Nature of breaks inducible in DNA at the early stages of interphase cell death]. AB - Dynamics of changes in 3'-OH- and 5'-OH-ends of DNA was determined by "nick" translation and direct polynucleotide kinase reaction, respectively, in animal thymocytes after irradiation and administration of hydrocortisone. Breaks bearing both 3'-OH- and 5'-OH-ends were found in DNA after irradiation. In 40 min repair of single-strand breaks was almost completed, and enzymatic breaks were accumulated with 3'-OH-ends only. 60 min after the administration of hydrocortisone, the number of nuclear DNA breaks containing 3'-OH-ends, but not 5'-OH-ends, sharply increased. Upon DNA autolysis in isolated nuclei acid nuclease produced 5'-OH-ends, and Ca2+/Mg2+-dependent nuclease, 3'-OH-ends. No activity of Mg2+-dependent nuclease was registered either in the nuclei of control thymocytes or in the nuclei isolated from thymocytes of exposed rats. PMID- 3033734 TI - [Effect of secondary radiation from 70-GeV protons and gamma quanta on chromosomes of Chinese hamster cells as dependent on the cell cycle stage]. AB - In cultured Chinese hamster cells, no decrease in the number of chromosome aberrations was noted after exposure thereof to 70 GeV protons at the late S phase as opposed to early one. It is suggested that high biological effectiveness of this type of radiation is associated with its inhibiting effect of cytogenetic damages repair. PMID- 3033736 TI - [Animal experiment study of the use of radioisotopes in studies on the hepatobiliary system. 1]. PMID- 3033735 TI - [Nonparametric method of determining the RBE coefficients of accelerated charged particles by the incidence of neoplasms in rats]. AB - The nonparametric method was used to determine RBE coefficients of accelerated charged particles (helium ions of 4 GeV/nucleon and 645 MeV protons) by the incidence of tumors localized in different rat organs or by the absence of tumors. The nonparametric method permitted to find the dose dependence of the RBE coefficients and to make statistical analysis of the results obtained with due regard for come features of developing damages which were not revealed by conventional methods of determining RBE coefficients. PMID- 3033738 TI - Glomus tympanicum chemodectomas: radiographic and clinical characteristics. AB - Glomus tympanicum chemodectomas are benign neoplasms that develop from normal glomus bodies located along the Jacobson (tympanic) nerve in the middle ear. The medical charts and radiographic studies of 55 patients with these tumors were reviewed. Women outnumbered men in a ratio of 3.5:1, and the patients' average age when they initially reported symptoms was 52 years. Tinnitus, ear pulsations, and diminished hearing were the most frequent symptoms. No patient had a second chemodectoma, and none of seven patients who were tested had elevated neuroendocrine compounds. Review of the radiographic examinations showed that direct coronal, thin-section computed tomography (CT) was the most sensitive means of demonstrating glomus tympanicum chemodectomas. Magnification angiography was also a sensitive diagnostic study, typically depicting a trapezoidal, hypervascular, middle-ear mass that appeared initially in the middle-to-late arterial phase and quickly disappeared in the venous phase. Differentiation from an aberrant internal carotid artery is critical to prevent arterial biopsy. PMID- 3033737 TI - Radioprotective effect of WR-2721 (gammaphos) or cystamine in non-uniform lethal gamma irradiated mice. PMID- 3033739 TI - Detection of epithelial- and neural type of intermediate filament proteins in human lung tumors. AB - Five different types of lung cancers, i.e. squamous cell carcinomas, adenocarcinomas, small cell lung carcinomas, carcinoids and adenoid cystic carcinomas were examined for their intermediate filament constituents, with special emphasis on the different cytokeratin polypeptides and neurofilament proteins. Polyclonal as well as monoclonal antibodies to these proteins were used in immunocytochemical techniques applied to both tumor frozen sections and paraffin sections. Squamous cell carcinomas and adenocarcinomas could be shown to contain cytokeratins, which could be detected in both frozen sections and paraffin sections. Also small cell lung carcinoma (SCLC) and carcinoid lung tumors showed a positive staining reaction with polyclonal and monoclonal (cyto)keratin antibodies, but were negative with neurofilament antibodies, with the exception of one case of lung carcinoid, which co-expressed neurofilaments and cytokeratins. We have used antibodies to cytokeratin polypeptides, to neurofilament proteins and to a neuroendocrine related membrane antigen (MOC-1) to further subclassify heterogeneously composed squamous cell carcinomas. Using a monoclonal antibody to cytokeratin 18, normally present in glandular tissues and adenocarcinomas, we observed that more than 90% of the squamous cell carcinomas examined can be stained with this antibody. The percentage of tumor cells, however, positive for cytokeratin 18 varies between 1 and 100%. In these same tumors a monoclonal antibody to skin keratins, which is known to react specifically with keratinizing cells, also stained variable numbers of tumor cells. This finding confirms the presence of (keratinizing) squamous cell carcinoma elements in these tumors. Our data show that most lung tumors, heretofore considered pure squamous cell carcinomas, should be considered biologically adenosquamous carcinomas. Also areas positive with MOC-1 were found in these tumors, suggesting the presence of squamous cell carcinomas with neuroendocrine differentiation. Furthermore, in some poorly differentiated squamous cell carcinomas areas with neurofilament positive cells were detected, suggesting a neural differentiation within these neoplasms. Adenoid cystic carcinomas are shown to co-express cytokeratins and vimentin in the tumor cells. This phenomenon can be used to identify such tumors and to distinguish them from other lung tumors. PMID- 3033740 TI - Immunohistopathologic spectrum of the glial fibrillary acidic protein in neurooncology. AB - To conventional histologic and special staining techniques as well as electron microscopy, immunohistological methods have recently been added to characterize neuroectodermal and non-neuroectodermal tumors of both the central and the peripheral nervous system. An ever increasing panel of these morphologic techniques enables us today, to characterize precisely such tumors in or around the central and peripheral nervous system, sometimes only hampered by preservation of tissue inadequate for the respective methods. PMID- 3033741 TI - Cortical neurochemistry in Alzheimer-type dementia. PMID- 3033742 TI - Neuronal cell membranes and brain aging. PMID- 3033743 TI - Neuropeptides and the treatment of cognitive deficits in aging and dementia. PMID- 3033744 TI - Endotracheal and endobronchial tumours in childhood. AB - Following a review of the literature on the most common types of endobronchial tumours (carcinoid cylindromas and mucoepidermoid tumours), four cases of such bronchial tumours in children are reported. Two of them underwent isolated bronchial resection, whereas one of the others had to undergo bilobectomy in addition and the other, pneumonectomy. The patients remained free of recurrences after a follow-up of 5-20 years. The advantage of circumferential resection and some technical aspects are discussed. PMID- 3033745 TI - [Physiological significance of the calpain-calpastatin system]. PMID- 3033746 TI - [Calcium-dependent proteases]. PMID- 3033747 TI - [Structure and function of calcium-activated neutral protease]. PMID- 3033748 TI - Depression and excitation induced in mice by two geometric isomers of (+)13,14 didehydro-20-methyl-carboprostacyclin, FCE 22177 and FCE 22176. Comparison with effects on blood pressure and platelet aggregation. AB - Two geometric isomers of a stable derivative of PGI2 (13,14-didehydro-20-methyl carbo PGI2), FCE 22177 (5E) and FCE 22176 (5Z), have been injected i.v. in mice (5-400 mcg/Kg) to investigate the effects at CNS level (Irwin's test). The synthetic mixture of the two isomers (5E:5Z = 7:3) was also tested. Isomers 5E and 5Z induced opposite effects: 5E (similar to PGI2) was depressive, 5Z induced stimulation. The mixture showed a strong depressive symptomatology. The two isomers and their mixture induced motor incoordination and impaired the performance of mice in the rotarod test (200 mcg/Kg i.v.). Mice were depressed after 5E and mixture; whilst 5Z induced excitation, in accordance with Irwin's test. 5E and 5Z showed analogous effects on mean blood pressure (hypotension) and heart rate (increase) in normotensive rats, but 5E was 7 times more active than 5Z. The two isomers showed analogous inhibitory effect on guinea-pig platelet aggregation in vitro, but 5E was 30 times more active than 5Z. In conclusion, the two geometric isomers have similar effects on blood pressure, heart rate and platelet aggregation, even if differing quantitatively. In contrast, 5E and 5Z have opposite effects at CNS level on motor function and behaviour. The data suggest the presence of two different sites of action for PGI2 in CNS (Ca2+- and adenylate cyclase-linked?). The two geometric isomers, 5E and 5Z, may be useful to study the PGI2 receptor sites in different tissues. PMID- 3033749 TI - Stimulation by melittin of adrenocorticotropin and beta-endorphin release from rat adenohypophysis in vitro. AB - The effect of melittin on the release of adrenocorticotropin (ACTH) and beta endorphin from the corticotropic cells of the rat adenohypophysis was examined in vitro. Anterior pituitary quarters were perifused or incubated in vitro and ACTH- (ACTH-IR) or beta-endorphin-like immunoreactivity (beta-End-IR) in the medium was measured by radioimmunoassays. Melittin stimulated ACTH-IR and beta-End-IR release. This effect was rapid in onset, reversible, and concentration-related (50-5000 ng/ml) and depended on the presence of calcium ions in the incubation medium. Melittin also elevated the tissue content of unesterified 3H-arachidonic acid that had previously been incorporated into lipids. Purported phospholipase A2 inhibitors, mepacrine (up to 1 mM), dexamethasone (0.5 mg/kg in vivo, 50 nM in vitro), or p-bromophenacylbromide (100 microM), did not decrease the melittin (500 ng/ml) - induced beta-End-IR release, although mepacrine and dexamethasone may have inhibited phospholipase A2 activity as indicated by an inhibition of melittin-evoked prostaglandin E2 formation. After stimulation by melittin (500 ng/ml), beta-End-IR release was not affected by the cyclooxygenase inhibitor indomethacin (up to 140 microM), whereas nordihydroguaiaretic acid (100 microM), a lipoxygenase inhibitor, or BW755C (250 microM), an inhibitor of both cyclooxygenase and lipoxygenase, abolished melittin-induced hormone secretion. We conclude that melittin generates a signal in the corticotropic cells of the rat adenohypophysis which induces hormone secretion by exocytosis. This signal may be unrelated to the activation by melittin of phospholipase A2. PMID- 3033750 TI - Cerebral hemisphere astrocytoma: treatment results. AB - Eighty two adult patients with histologically proven cerebral astrocytomas of grades I to IV received post-operative radiotherapy at Westmead Hospital between January 1980 and February 1985. The extent of surgery consisted of biopsy alone in 44%, subtotal tumour resection in 48%, and "complete" tumour removal in 8%. Seventy one patients completed a course of megavoltage irradiation, the majority having received a tumour dose of at least 60 Gy. Patients who underwent surgical resection (complete or incomplete) had a greater median survival (14 months) than those undergoing biopsy (8 months), but the difference was not statistically significant (p = 0.08). By grade, the difference reached statistical significance only for grade III tumours (p = 0.015). Patients with high grade tumours had a significantly lower survival than those patients with tumours of low grade. Median survival for patients with grades I and II, III and IV tumours was 42.0, 12.0 and 7.0 months, respectively. After adjustment for grade, various dosage levels (less than 60, 60 or greater than 60 Gy) did not significantly affect survival, although there was a trend towards improved median survival with higher doses in grade III tumours. Older patients (greater than 45 years) had a significantly lower median survival (25 months) than younger patients (8 months) (p less than 0.0001). When included in a multivariate analysis, the extent of surgery did not significantly influence survival, but increasing tumour grade and increasing age were significant adverse prognostic factors. PMID- 3033751 TI - Quality of life of inoperable non-small cell lung carcinoma. A randomized phase II clinical study comparing radiotherapy alone and combined radio-chemotherapy. AB - Eighty one patients with inoperable non-small cell lung carcinoma (NSCLC) were entered in a randomized phase II trial comparing split-dose irradiation alone to combined treatment radiotherapy and polychemotherapy (C.A.P. + V.D.S.). The quality of life and the survival of the patients were studied. We have defined three classes of quality of life responses based on the time elapsed before the performance status index drops. A higher quality of life failure rate was observed in the combined treatment group (p non-significant) but the time elapsed before the Karnofsky index drops is longer in the combined treatment group for the quality of life "no change" subgroup (p = 0.15). Survival and quality adjusted survival are similar in both treatment groups. The same conclusion holds for retrospective stratified treatment groups. The results of the study are presented according to the decision tree theory. We conclude that as far as the quality of life is concerned, polychemotherapy combined with the particular split dose irradiation schedule used is an effective treatment of inoperable NSCLC. Its efficiency is comparable to, but not better than, the same radiotherapy schedule alone taken as a reference baseline. PMID- 3033752 TI - Effect of kassinin, neurokinin A and neurokinin B on drinking behaviour in the pigeon. AB - Intracerebroventricular (i.c.v.) injection of kassinin produced a prompt and copious drinking response at doses of 10-1000 ng/pigeon, in the absence of other behavioural alterations or of changes in core temperature. Neurokinin A and B evoked drinking, but they were respectively 10 and 100 times less potent than kassinin. Intraperitoneal injection of kassinin elicited drinking, but at doses about 1000 X larger than the i.c.v. ones. The angiotensin antagonist [Sar1, Leu8]angiotensin II did not reduce drinking induced by i.c.v. kassinin, suggesting that its effect is not due to interaction with the central renin angiotensin system. Moreover, the effect is apparently independent of the mechanisms controlling hypovolaemic and hyperosmotic thirst since exact additivity was found in the dipsogenic response when i.c.v. kassinin was administered in the presence of a hypovolaemic (subcutaneous (s.c.), polyethylene glycol) or hyperosmotic (s.c. hypertonic NaCl) dipsogenic stimulus. The present findings show that kassinin, neurokinin A and B share with the tachykinins already tested (eledoisin, physalaemin, substance P) a common dipsogenic action in pigeons. However, marked differences exist in their dipsogenic potency. This order of potency, eledoisin = kassinin = physalaemin greater than neurokinin A = substance P greater than neurokinin B, is not consistent with the tachykinin receptor subtypes so far proposed. PMID- 3033753 TI - [Clinical evaluation of serum neuron-specific enolase levels in patients with lung cancer]. AB - Serum neuron-specific enolase (NSE) levels were studied in 105 patients with malignant neoplasms (lung cancer 38, others 67), 13 patients with various benign diseases and 7 healthy adults. The mean serum NSE level in adult control subjects was 7.4 +/- 0.8 ng/ml, and cut off level was decided 10 ng/ml. Serum NSE levels were elevated in 14/38 (37%) of patients with lung cancer and in 14/67 (21%) of patients with the other malignant neoplasms. In patients with benign diseases, serum NSE level was elevated only in one patient with pituitary adenoma. In 7 patients with small cell lung cancer, the positive rate was higher (86%) than in those with non-small cell lung cancer (26%), and serum NSE levels were higher than 25 ng/ml except one case. There was no correlation between serum NSE and CEA (carcinoembryonic antigen) levels in patients with small cell lung cancer, also in patients with lung cancer. The measurement of serum NSE level seemed to be useful for diagnosis in patients with small cell lung cancer. PMID- 3033755 TI - [Non-functioning pancreatic islet cell tumor treated by total pancreatectomy]. PMID- 3033754 TI - [Gestational trophoblastic neoplasms]. PMID- 3033757 TI - Computed tomography of bilateral carotid body tumours. AB - The CT images in 4 patients with bilateral carotid body tumours are reported. These CT findings seem to be typical, consisting of a well-circumscribed and strongly contrast-enhancing mass, situated at the bifurcation of the common carotid artery, displacing the adjacent structures. Computed tomography proves to be an excellent and non-invasive diagnostic examination. Angiography is necessary only if embolisation prior to surgery is requested. PMID- 3033758 TI - [Computed tomography of anterior dislocation of the temporomandibular articular disk]. AB - In a prospective study, the findings on arthrography and CT in 51 TMJs with anterior meniscus dislocation were compared. The CT diagnosis of anterior dislocation was made by a standardised method which we have developed and which depends upon evaluating axial sections. Sensitivity of the CT methods was 87%. In the absence of arthrography, a negative diagnosis in the asymptomatic contralateral side was made in 81%. Ruptures and adhesions could not be demonstrated by CT. For these reasons, arthrography is to be preferred to CT for making the initial diagnosis. CT should be carried out for special indications, such as contrast sensitivity, the documentation of a clinically confirmed diagnosis and for follow-up of treatment. Currently an attempt is being made to stage the disease by computed tomography. PMID- 3033759 TI - [Value of high-resolution CT and nuclear magnetic resonance tomography compared to the standard procedures in the diagnosis of meniscal lesions]. AB - The knees of 20 patients with evidence of meniscal tears were examined via high resolution computed tomography (HRCT); 10 of these were studied by MRI. The HRCT study was performed directly after double-contrast arthrography (AG). For comparison with HRCT, slice orientation for MRI examination was in transverse view; gradient echo sequences using the FISP technique were applied instead of spin echo sequences. All results were correlated to the arthroscopy (AS) findings. In 95% of the cases AG and AS results agreed, HRCT/AS in 85% and MRI/AS in 70%. In certain cases HRCT provided additional information which influenced appropriate surgical treatment. MRI is a noninvasive nonionising method but gives a less exact documentation of the lesion than AG and HRCT. The gradient echo mode is superior to the SE mode in respect of outlining meniscal structures, at least in transverse view. PMID- 3033756 TI - [Technic and results of computed tomography of the base of the nose and orbit with multiplanar reconstructions]. AB - The additional use of a 3-D display programme provides valuable information in cases of trauma to the mid-face and tumours of the bony orbits. The computer programme provides important additional information for the radiologist and, in future, will influence treatment significantly. PMID- 3033760 TI - High resolution MR imaging of peripheral joints using a quadrature coil at 0.35 T. AB - The utility of a specially dedicated quadrature detection (QD) mode limb coil for the MR evaluation of peripheral joints has been evaluated. High resolution images of the knee, elbow, wrist and ankle provided excellent anatomical delineation of the cortical and medullary bone, cartilage, ligaments and menisci over a homogeneous field of view. Pathological conditions including meniscal injury, fracture and osteonecrosis were accurately detected. The QD limb coil can be advantageously used for the MR evaluation of peripheral joints at an intermediate field strength. PMID- 3033761 TI - [Computed tomography of intra-articular calcaneal fractures]. AB - The increasing number of operations on intraarticular calcaneal fractures created the need for a fast, reproducible method to examine the weight-bearing posterior talo-calcaneal joint. High resolution CT scanning in an almost coronal plane without reformating was done in 25 calcaneal fractures. Joint alignment and fragment dislocation were demonstrated more precisely than would have been possible with conventional studies. Three typical fractures were found: The lambda type, the y type and the comminution type. Follow-up studies showed osteo arthritis, intraarticular degenerative changes of the articular surface and impingement of personeal tendons as possible causes of pain. PMID- 3033762 TI - [Space-occupying tumor lesions of the spinal column. An analysis of the material submitted to the Westphalia Bone Tumor Register]. AB - 308 tumorous lesions of the spine were identified in the material submitted to the bone tumour register in Munster for recording. Histological examination revealed metastases in 38.6% primary bone tumours in 33.1%, tumour-like lesions in 10% and haematological and lymphatic disease in 13%. 5.3% of all lesions could not be classified in these groups. Primary bone tumours were seen more often than metastases in the cervical spine, the sacrum and the coccyx. 54.9% of the primary bone tumours of the spine were benign. However, only 31.2% of all the tumorous lesions of the spine, metastases included, were benign. The portion of malignant disorders increased with increasing age. The risk of having a malignant spine tumour was 5.9% in patients under 10 years of age but 92.3% in patients in the seventh decade. PMID- 3033763 TI - [Diagnostic value of ultrasound in malignant melanoma]. AB - High-resolution real-time sonography enables visualisation of the morphology of the cutis and of cutaneous tumours. Evaluation of 26 malignant melanomas showed that there is a high degree of correlation between the sonographically measured values of maximal tumour thickness with those determined postoperatively by histometry. As malignant melanomas have very few internal echos, they can be easily differentiated from benign tumours. High-resolution sonography is thus the only diagnostic imaging method which helps to evaluate preoperatively malignant melanomas. PMID- 3033764 TI - [Prostatic cancer. Staging via transrectal prostatic sonography and computed tomography with histopathological correlation]. AB - Pre-operative staging, using transrectal prostatic sonography and CT, was carried out in 30 patients with cytologically confirmed carcinomas of the prostate and the results compared with the clinical findings. All patients underwent radical prostatectomy and the pre-operative findings could be verified histologically. Transrectal prostatic sonography is better than CT or clinical examination for determining local tumour spread or penetration of the capsule. A high proportion of enlarged pelvic lymph-nodes shown by CT had non-specific changes; failure to demonstrate enlarged nodes excludes lymph node metastases with considerable certainty. Transrectal prostatic sonography provides a higher degree of information regarding local tumour spread, whereas CT indicates the presence or absence of lymph node metastases. PMID- 3033765 TI - [The internal echo pattern of the normal pancreas. An assessment of the aging process and body weight dependence by sonographic gray-scale analysis]. AB - Sonography of the pancreas was evaluated quantitatively in 94 normal subjects. The grey-scale values in the head and body of the pancreas were measured and compared with those of retroperitoneal fat. The echogenicity of the pancreas and the contrast between pancreas and fat were correlated with the age and weight of the subjects. Semi-quantitative studies revealed a positive correlation between echogenicity and age (r = 0.505); this can be confirmed to a high degree of statistical significance by quantitative analysis of pancreatic grey-scale values (r = 0.55 for the head and 0.71 for the body). Further improved correlation with age is obtained by computing the sonographic contrast between the pancreas and the retroperitoneal fat (r = 0.66 for the head and 0.77 for the body of the pancreas). It is concluded that determination of the grey-scale value can provide additional information particularly if one uses retroperitoneal fat as a reference tissue. PMID- 3033766 TI - [DSA for the imaging of tumor-caused vascular changes and complications during local chemotherapy of the liver via the portal system]. AB - In 18 patients with primary or secondary liver tumours, 38 digital subtraction angiographies (DSA) of totally implanted catheter systems for liver perfusion were performed during a period of 8 months. The systems had been used for an average of 6.9 months. In 22 out of 38 examinations secondary reactions of liver arteries were observed, in 7 the therapeutic regimen was changed after DSA. Patients with an extensive tumour involvement of the liver showed mural thromboses or obstructions of liver arteries relatively more often (4 of 7) than patients with smaller size tumours (2 of 11). In 7 cases the therapeutic regimen was changed after DSA. DSA of implanted liver catheters is a valuable, well tolerated, easily and rapidly performed method. PMID- 3033767 TI - [Potentials of nuclear magnetic resonance tomographic diagnosis of the adrenals. Initial experience using a Helmholtz surface coil]. AB - In order to obtain optimum spatial resolution in the adrenal glands, images were obtained using Helmholtz surface coils and respiratory gating. In addition to normal controls, 9 patients with space-occupying lesions were examined (4 metastases, 3 adenomas, 2 phaeochromocytomas). It was found that tissue-specific appearances were associated only with phaeochromocytomas, depending on the high signal intensity on their T2 sequences. PMID- 3033768 TI - [Magnetic resonance tomography of ovarian tumors]. AB - The results of 64 MR examinations in 52 patients with benign, malignant and inflammatory lesions of the adnexa are presented. In each case, surgery or laparoscopy was carried out, so that it was possible to compare the findings on MRT with the histological results. Multiplanar images of gynaecological lesions provide excellent demonstration of the pathological anatomy. High soft tissue contrast makes it possible to differentiate tissues, particularly in the case of serous, mucinous and haemorrhagic fluid collections, as well as solid and fat containing tissue. High sensitivity may be expected in the demonstration of dermoid and endometrial cysts. The distinction between benign and malignant lesions must, however, be based on morphological criteria. The combination of T1 and T2 sequences is necessary for adequate MR examinations of gynaecological lesions. PMID- 3033769 TI - [Digital subtraction angiography and large-film angiography of the pelvic and leg arteries]. AB - The diagnostic information and image quality of 1153 DSA studies of pelvic and lower limb arteries and of 104 conventional large film angiograms have been analysed. Using intra-arterial contrast, all large film angiograms and all DSA studies of the iliac, femoral, popliteal and upper calf arteries were satisfactory. For demonstrating the distal calf vessels and vessels in the foot, intra-arterial DSA was superior to large film angiography. Using intravenous DSA, 98% of the iliac, femoral and popliteal arteries were adequately demonstrated, although image quality was reduced. PMID- 3033770 TI - Sequential PTA of abdominal aorta. Haemodynamic evaluation and IV-DSA follow-up. AB - A case of sequential dilatation of a subtotal stenosis of the abdominal aorta in a young subject is reported. Initial and long-term success of the procedure is recorded using haemodynamic evaluation and intravenous digital subtraction angiography (IV-DSA) follow-up on an outpatient basis. In addition, the significance of biplane aortography with IV-DSA is illustrated. PMID- 3033772 TI - [Determination of the integral doses in conventional x-ray studies and computed tomography of the skull]. AB - Absorbed energy (integral dose) is a suitable measure of radiation exposure during radiodiagnostic examinations. The integral doses from conventional and CT examinations of the skull were determined, using an Alderson-Rando phantom. The absorbed energy of one exposure using a film-screen combination and for one plane of conventional tomogram is almost identical. Considering the total absorbed energy, lower doses are to be expected for CT than for conventional tomography. A survey view is associated with reduced radiation exposure. PMID- 3033771 TI - [Initial experiences with CO2 as a gaseous contrast medium in digital subtraction angiography]. AB - Electronic contrast enhancement allows the use of CO2 as a contrast material for studying the vascular system with DSA. Its use, safety and the information provided were studied in 40 patients and in 5 animal experiments and the results were compared with iodine-containing contrast media. The available results indicate that CO2 is a safe negative contrast medium for peripheral arteriography and that it provides angiograms with a picture quality comparable to those of iodinated contrast media in the extremities. The method can therefore be used to advantage in patients with allergies to the iodinated contrast media and in patients with renal failure. PMID- 3033773 TI - [Making a diagnosis in postpartal ovarian venous thrombosis by digital subtraction angiography]. PMID- 3033774 TI - Solitary osteochondroma of the spine. Report of two unusual cases in children. PMID- 3033775 TI - [Cholesterol granuloma (so-called cholesteatoma) of the breast. The differential diagnosis of a slowly growing cancer]. PMID- 3033779 TI - [68th German Roentgen Congress. Film interpretation seminar. 1 May 1987]. PMID- 3033776 TI - [Cavernous hemangioma of the renal pelvis]. PMID- 3033777 TI - [Congenital aneurysm of the sinus of Valsalva with perforation into the right ventricle]. PMID- 3033778 TI - [Ergotismus as a consequence of migraine prevention]. PMID- 3033780 TI - Plasma renin activity and aldosterone concentration in sodium-depleted cattle following ACTH or metoclopramide injection. AB - Adrenocorticotropic hormone (ACTH) treatment (7 micrograms X kg-1 body weight) induced an increase in plasma aldosterone concentration in both Na-deficient heifers (following saliva loss after right parotid duct cannulation) and Na replete heifers, but had no significant effect on plasma renin activity in either of these groups. The dopamine antagonist, metoclopramide, injected i.v. (1 mg X kg-1 body weight) did not modify plasma renin activity, aldosterone or cortisol concentrations in either group. These results indicate that dopamine did not play a major role in the regulation of aldosterone secretion in Na-replete or Na depleted heifers. PMID- 3033781 TI - Quantitative structure-activity relationship study on hexestrol and metahexestrol derivatives interacting with estrogen receptors. AB - The estrogen receptor binding affinity of various hexesterol derivatives is found to be significantly related with van der Waals volume of substituents and the Hammett constant sigma. This led to suggest that substituents produce steric effects on the drug-receptor interaction and that the electron-donating capability of substituents enhances the activity. In case of metahexestrol derivatives these factors were however not found to affect the binding affinity of compounds. The reason thereof is discussed. PMID- 3033782 TI - Effect of ACE-inhibition on tissue blood flow during acute left ventricular failure in the dog. AB - Anesthetized dogs in acute left ventricular failure were treated with the ACE inhibitor enalaprilat (MK-422). ACE-inhibition produced a fall in the mean blood pressure and a redistribution of cardiac output to the brain, right ventricle, upper gastrointestinal tract, and the inner and middle part of the renal cortex. The flow to the spleen, adrenals, skin, muscle, fat, lower gastrointestinal tract, and outer renal cortex did not change significantly. The differential sensitivity of the tissues to ACE-inhibition is most likely due to differences in sensitivity to circulating angiotensin II. PMID- 3033783 TI - Biochemical indicators of disordered vitamin D and calcium homeostasis in sarcoidosis. AB - In an attempt to identify factors that may indicate which patients with sarcoidosis are likely to manifest a clinical abnormality in calcium homeostasis, we measured serum concentrations of calcium, angiotensin converting enzyme activity (ACE), 25-hydroxyvitamin D (25-OH-D), 1,25-dihydroxyvitamin D (1,25 (OH)2-D), and immunoreactive parathyroid hormone (iPTH) in 19 patients with biopsy proven sarcoidosis, seven of whom were either frankly hypercalcemic or hypercalciuric. These data were compared to the ability of cultured pulmonary alveolar macrophages (PAM) from the same patients to metabolize [3H]25-OH-D3 to [3H]1,25-(OH)2-D in vitro. All seven hypercalcemic/hypercalciuric patients with sarcoidosis had a serum ACE level greater than 40 IU/L (normal less than 35 IU/L). All five patients with hypercalcemia (Ca greater than or equal to 10.5 mg/dl) had a serum 1,25-(OH)2-D concentration above the normal range (greater than 60 pg/ml), and in all 19 patients the serum calcium was positively correlated to the serum 1,25-(OH)2-D concentration (r = 0.55, p less than 0.01). The capacity of PAM to synthesize [3H]1,25-(OH)2-D3 in vitro was. with one exception, greater in cells from patients with diffuse infiltrative pulmonary disease (roentgenographic stage II or III) and positively correlated to the serum calcium concentration (r = 0.72, p less than 0.001) and serum ACE (r = 0.43, p less than 0.05). Cultured PAM from the five hypercalcemic patients, all of whom demonstrated diffuse pulmonary disease on chest x-ray, showed a [3H]1,25-(OH)2-D3 synthetic capacity in vitro 2.5-fold greater than that for group as a whole and 6 fold greater than in cells from nonhypercalcemic patients with sarcoidosis. PMID- 3033784 TI - Comparison of clinical parameters, bronchoalveolar lavage, gallium-67 lung uptake, and serum angiotensin converting enzyme in assessing the activity of sarcoidosis. AB - A major problem in pulmonary sarcoidosis is the assessment of disease activity. We established a clinical score system composed of degree of dyspnea, chest radiographic stage, forced vital capacity, and clinical assessment of disease activity (normal = 0, maximum = 16). We evaluated this score in 50 patients with sarcoidosis along with other proposed measures of disease activity: bronchoalveolar lavage percent lymphocytes (% lymphs), Gallium-67 lung uptake measured as total lung to background ratio (TL/B ratio), and serum angiotensin converting enzyme (SACE) levels. The patients we studied had an average clinical score of 5.6 +/- 2.7 (+/- SD). Though bronchoalveolar lavage % lymphs (32 +/- 20%), TL/B ratio (192 +/- 73), and SACE (137 +/- 66) were all significantly elevated in these patients, Gallium-67 uptake was most frequently abnormal (80% of patients). However, the TL/B ratio was higher in smokers (231 +/- 91) than nonsmokers (178 +/- 64, p less than 0.05). There was no correlation between our clinical score, or any part of the clinical score, and other laboratory measures of alveolitis. Bronchoalveolar lavage % lymphs and TL/B ratio correlated weakly (n = 50, r = 0.36, p less than 0.02), while % lymphs and SACE correlated less well (n = 29, r = 0.36, p = 0.051). There was no correlation between TB/L ratio and SACE.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3033785 TI - The significance of angiotensin I converting enzyme in granulomatous inflammation. Functions of ACE in granulomas. AB - ACE is a carboxy-terminal peptidyl dipeptidase which can be produced by granuloma macrophages and possibly by other components of the granuloma. ACE activity is influenced by regulatory T cells. This enzyme may be important in mediating and/or modulating inflammation. One mechanism involves the generation of angiotensin II which can effect inflammatory reactions. Although unstudied, ACE may also be an important inactivator of bradykinin and various neuropeptides which in turn may participate in the immunoregulation of the granuloma. Thus, therapeutic strategies designed to alter ACE activity within granulomas may alter the size, composition and ultimate resolution of these lesions. PMID- 3033786 TI - Production of the angiotensin converting enzyme (ACE) in cultures of rat peritoneal macrophages. AB - Some authors attributed an increase in ACE activity in sarcoidosis to high production of this enzyme by alveolar macrophages. In order to study the production of ACE by peritoneal rat macrophages, we measured the ACE activity, by a modified version of Friedland and Silverstein's technique, in cultures of rat peritoneal macrophages, with dexamethasone, with Complete Freund Adjuvent (C.F.A.) or sarcoidosis node extract (Kveim Antigen). ACE activity was not detected in cultures. Our results were discussed. PMID- 3033787 TI - Immuno-cytological blood tests in cases of sarcoidosis. AB - Monocytosis, occurring in the majority of sarcoidosis cases in the bloodstream, and an increased number of natural killer cells are a manifestation of an increased cellular defence by bone marrow cells. The antigen-offering function of natural killer cells, macrophages and monocytes, should be specially emphasized. While the majority of sarcoidosis patients show an absolute lymphocyte count of less than 1500 cells/microliter during the usual clinical blood tests, the determination of lymphocyte subpopulations by means of monoclonal antibodies displayed a different behaviour of the lymphocyte subpopulations, especially a substantial reduction of T-cells, while B-lymphocytes were at normal levels. The assumption, that sarcoidosis immunology is characterized by an imbalanced homeostasis in the sense of a decrease in cellular and an increase in humoral immune defence, can no longer be maintained. We are rather dealing with an increased immunological activity of the cellular and humoral immune defence. In addition, the investigation of the lymphocyte subpopulations in the blood allows a judgement on the existing activity of the disease. The statement of a helper suppressor ratio of less than 2.0 suggests progress of disease and indicates therefore drug therapy. Our immunological findings also explain the difference of the prognosis, which is more favourable for women than for men. PMID- 3033789 TI - The Italian approach to sarcoidosis and other granulomatous disorders. PMID- 3033788 TI - Serum angiotensin-converting-enzyme in rabbits with and without pulmonary granulomatosis. Granulomatosis induced with complete-Freund's-adjuvant or BCG. AB - An animal model with pulmonary, non-caseating, granulomatosis was successfully reproduced in rabbits utilizing intravenous injections of complete-Freund's adjuvant (CFA) or BCG. Granuloma formation from CFA injections was limited to the lungs, but BCG injections caused systemic granulomatosis (lung, liver and spleen) irrespective of whether the BCG was dissolved in saline or in incomplete-Freund's adjuvant (IFA). Serum angiotensin-converting-enzyme (ACE) levels began to rise approximately 15 days following the intravenous injection of CFA, but similar rises following the injection of IFA without granuloma formation suggest that the ACE rise was not necessarily reflective of granuloma formation in rabbits. BCG in saline caused no rise in serum ACE, but when suspended in IFA the ACE rise was comparable to that resulting from IFA alone. However, the ACE level of broncho alveolar-lavage cells obtained from rabbits 21 days post-injection of CFA was 7 fold higher than that of lavaged cells obtained 42 days post injection, correlating with an acute alveolitis occurring early during granuloma formation. These studies confirm the usefulness of intravenous injections of CFA or BCG for inducing a granulomatous state, but show that the assay of serum ACE is not useful for reflecting the granulomatous state in rabbits. In addition, serum ACE levels are 2-5 X higher in rabbits than in humans and tend to fluctuate spontaneously following delivery of the animals to the laboratory. PMID- 3033790 TI - [Myocardial scintigraphy with Tc 99m-pyrophosphate in patients with acute transmural myocardial infarction, non-transmural infarction and unstable angina]. PMID- 3033791 TI - [Infection by HTLV III virus in Chilean homosexuals of median and high socioeconomic levels]. PMID- 3033793 TI - [New data regarding multiple sclerosis]. PMID- 3033792 TI - [Determination of free radicals by ESR in lyophilized tissues]. PMID- 3033794 TI - [The case from practice (76). Female patient: Mrs. P. R., born 3-16-1920, homemaker]. PMID- 3033795 TI - [Virus-host relations: from confrontation to peaceful coexistence]. PMID- 3033797 TI - [Functions of the bronchoalveolar cells]. PMID- 3033796 TI - [2 cases of Waldenstrom's disease associated with a glioblastoma]. AB - The association of Waldenstrom disease and glioblastoma was observed in two patients. Both diseases were diagnosed almost simultaneously. In the first patient the diagnosis of glioblastoma was made on cerebral biopsy. In the second case, the diagnosis was confirmed at autopsy. The association of Waldenstrom's disease and glioblastoma is rare but it does not seem to be coincidental. The physiopathological problems related to this coexistence are discussed. PMID- 3033798 TI - [Structure and functions of the interlobular spaces]. PMID- 3033799 TI - [New therapeutic strategies in bronchial asthma]. PMID- 3033800 TI - [Vepeside (VP-16-213) in the treatment of bronchopulmonary cancer (preliminary data)]. PMID- 3033801 TI - [Immune reactivity and immunomodulator-induced changes in bronchopulmonary infections]. PMID- 3033803 TI - [Critical aspects of the information capacity of a pleural needle biopsy]. PMID- 3033802 TI - [Current clinical characteristics of intestinal tuberculosis]. PMID- 3033804 TI - Morphological and ultrastructural changes of lung tissue in idiopathic fibrosing alveolitis. PMID- 3033805 TI - [Diagnosis and treatment of urinary and genital tuberculosis in men]. PMID- 3033807 TI - [Bacteriological diagnosis of intestinal tuberculosis]. PMID- 3033806 TI - [Acquired immunodeficiency syndrome (AIDS): the implications for current pneumological practice]. PMID- 3033808 TI - Effects of sodium and potassium adenosine-triphosphatase on circulating lymphocytes: an approach to human obesity. AB - The methodological aspects of (Na+, K+)-ATPase-dependent uptake of 86Rb, a potassium analog, were examined on human lymphocytes isolated from peripheral blood. The study of the time-course, the kinetic parameters, i.e., maximum velocity (Vmax) and Michaelis constant (Km) and the ouabain inhibition curve of 86Rb+ uptake confirm that circulating lymphocytes represent a suitable model for the study of (Na+,K+)-ATPase in human diseases. An application to human obesity is reported: the results indicate that 86Rb+ uptake on circulating lymphocytes is similar in obese and non-obese subjects. Therefore, (Na+,K+)-ATPase does not seem to be involved in the pathogenesis of human obesity. PMID- 3033810 TI - [Hematogenous gastrointestinal metastases--limits of radiologic diagnosis]. AB - Roentgen morphological, ultrasonographical and CT findings in a patient with mammary cancer metastasising hematogenously to the duodenum and colon and in four patients with malignant melanoma metastasising hematogenously to the stomach, duodenum, small bowel and colon. PMID- 3033809 TI - Alteration of DNA tertiary structure by physical and chemical carcinogens: involvement in DNA repair processes. AB - Parameters defining the topological state of DNA seem extremely important for describing the reactive state of the same DNA molecules. We have shown that physical and chemical DNA modifying agents alter the tertiary structure of DNA molecules. Variations in the tertiary structure of DNA were studied by one dimensional electrophoresis on an agarose gel of partially relaxed plasmid DNA topoisomers, a technique allowing the measurement of alterations in the degree of supercoiling equivalent to fractions of superhelical turns. Unwinding angles of 10.1 degrees or -8.7 degrees per pyrimidine or thymine dimer respectively, of -12 degrees per apurinic site, and of -3.4 degrees per methylated purine were obtained by titrating the number of damaged sites necessary to reduce the number of superhelical turns by one in each topoisomer. On the contrary, enzymatic methylation of the C-5 position of cytosine (a modified base present in prokaryotic and eukaryotic DNAs) did not alter the DNA tertiary structure. We have also shown that local alterations in DNA structure caused by UV-irradiation inhibit bacterial DNA topoisomerase I and DNA methylase, and that the topological state of DNA substrate influences the mode of methylation of Hpa II DNA methylase. These findings suggest that the natural topological state of DNA substrate (linear, relaxed, or covalently closed duplex DNA with varying degrees of supercoiling) influences the mode of action of enzymes possibly involved in DNA repair processes, while DNA structural alterations caused by DNA modifying agents might influence DNA repair processes in two ways: either by driving the interaction between repair enzymes and the modified sites of DNA, or by inhibiting or changing the mode of action of enzymes normally acting on unmodified DNA. PMID- 3033813 TI - Modulation of cholinergic nerve transmission by leucin5-enkephalin. PMID- 3033811 TI - Effect of calcitonin on phosphorylase a and glucose-6-phosphatase activities in the hepatic particulate glycogen of rats. AB - A single dose of calcitonin (CT) (70 mU MRC/100 g body weight) was administered in 50 and 90 days old male Wistar rats. In both ages CT determined a rise of Phosphorylase a and glucose-6-phosphatase activity as well as an increased calcium accumulation at the level of liver particulate glycogen. In the rats receiving CT glycemia increased and the liver glycogen content decreased. With the exception of liver glycogen the other metabolic parameters studied were modified at the two ages in the same manner under the influence of the same dose of CT. PMID- 3033812 TI - Adenohypophyseal hormone levels during hyperthermia. AB - The effect of hyperthermia on some adenohypophyseal hormone levels (prolactin, growth hormone, adrenocorticotropin, thyrotropin) in plasma was studied. The levels of prolactin increased significantly (310%) during Finnish sauna. The growth hormone levels increased slightly, whereas the levels of adrenocorticotropin and thyrotropin were unaltered. The increase in prolactin levels during hyperthermia resembled the changes in the levels of prolactin which have been found during stress and exercise. PMID- 3033814 TI - Subcellular localization of angiotensin-converting enzyme in the human alveolar macrophage. AB - The present investigation was performed in order to elucidate the subcellular localization of angiotensin-converting enzyme (ACE) in human alveolar macrophages. A pure population of alveolar macrophages was obtained by centrifugal elutriation of bronchoalveolar lavage (BAL) fluid from seven sarcoid patients. The cells were homogenized by sonication and the postnuclear supernatant was fractionated on a discontinuous sucrose gradient. Fractions of particulate material were collected and characterized by marker enzymes. The distribution pattern of ACE closely resembled that of NADPH-cytochrome-c reductase and sialyltransferase, markers of the endoplasmic reticulum and the Golgi complex, respectively, indicating a common localization. This localization is compatible with synthesis taking place in the alveolar macrophage. PMID- 3033815 TI - Short-chain fatty acids and the irritable bowel syndrome: the effect of wheat bran. AB - Short-chain fatty acids (SCFA) in faeces were examined in 18 patients with the irritable bowel syndrome (IBS) during treatment with wheat bran or placebo. In the placebo period, the patients could be classified in accordance with the faecal concentrations of SCFA into one group with low concentrations (mean, 40 mmol/l; range, 19-77 mmol/l; 10 patients) and another with high concentrations (mean, 168 mmol/l; range, 145-187 mmol/l; 8 patients). The concentrations of SCFA differed (P less than 0.001) in both groups from concentrations found in faeces from a reference group of nine normal individuals (mean, 114 mmol/l; range, 93 155 mmol/l). Patients with low levels of SCFA had lower (P less than 0.001) mean stool mass and longer (P less than 0.05) transit times than those with high concentrations of SCFA in faeces. Ingestion of bran, although a precursor of SCFA, did not change faecal concentrations of SCFA. Abdominal pain, distension, and rumbling were not correlated to low or high concentrations of SCFA in faeces, nor did bran improve these symptoms when compared to placebo. The level of SCFA was rather constant intraindividually and independent of the variability of the daily faecal mass. It is concluded that patients with IBS apparently have continuously abnormal concentrations of SCFA in faeces, either high or low, which are unaffected by the treatment with bran and which hypothetically may be of pathophysiologic importance. PMID- 3033816 TI - DCCD (N,N'-dicyclohexylcarbodiimide) inhibits biliary secretion of HCO-3. AB - To study whether a proton pump is an integral part of the mechanism responsible for secretin-dependent biliary secretion of HCO-3 ions, the proton pump inhibitor N,N'-dicyclohexylcarbodiimide (DCCD) was systemically administered to six anesthetized, secretin-infused pigs. Because biliary HCO-3 secretion varies with arterial pH, secretion rate was measured at several different arterial pH values, before and after DCCD (25 mumol/kg). At arterial pH 7.45, bile flow was 2.1 (1.6 2.9) ml/min, and HCO-3 secretion was 224 (157-311) mumol/min. DCCD reduced bile flow and HCO-3 secretion by 30% and 40%, respectively, independent of arterial pH. In contrast, bile acid secretion, 46 (41-59) mumol/min, was not changed by DCCD. The hepatic adenosine triphosphatase (ATP) level, 2.0 (1.8-2.1) mumol/g wet tissue, was not changed by DCCD. DCCD (10(-4) mol/l) affected neither Na,K-ATPase nor carbonic anhydrase activities in separate in vitro assay systems. The reduction in biliary HCO-3 secretion induced by the proton pump inhibitor DCCD may indicate that a proton pump is integrated into the mechanism responsible for secretin-dependent biliary secretion of HCO-3. PMID- 3033817 TI - Generation of reactive oxygen radicals by human phagocytic cells activated by Plasmodium falciparum. AB - The role of monocytes, macrophages and neutrophils in killing malaria parasites is well documented, and their involvement in malaria pathology has been suggested. However, the underlying mechanisms are not clear. The present study reports on the role of P. falciparum-parasitized erythrocytes, free merozoites, and culture supernatant antigens in the generation of reactive oxygen radicals by human peripheral blood monocytes and neutrophils. Blood neutrophils and monocytes obtained from healthy individuals were isolated by density gradient separation. A human isolate of P. falciparum was grown in continuous culture. Parasitized erythrocytes and free merozoites were prepared from synchronized cultures. Soluble antigens from culture supernatants were purified by affinity chromatography using CNBr-Sepharose 4B columns bound to specific IgG. Oxidative burst response of neutrophils and monocytes were determined by oxygen consumption, superoxide production, and chemiluminescence. It was found that P. falciparum merozoites and the soluble antigens were capable of activating neutrophils and monocytes in vitro and resulting in the production of oxygen radicals by these cells. In conclusion, these findings demonstrate that malaria antigens are able to activate normal human blood phagocytes and result in generation of oxygen radicals by these cells. The released oxygen radicals can then contribute to both the destruction of the parasite and the pathology of malaria. PMID- 3033818 TI - Cancer mortality of granite workers. AB - A retrospective cohort study was undertaken to investigate the cancer mortality of granite workers. The study comprised 1,026 workers hired between 1940 and 1971. The number of person-years was 20,165, and the number of deaths 235. During the total follow-up 46 tumors were observed and 44.9 were expected. An excess mortality from tumors was observed for the workers followed for 20 years or more, the greatest excess occurring during the follow-up period of 25-29 years (observed 11, expected 5.2). Of the 46 tumors, 22 were lung cancers (expected 17.1) and 15 were gastrointestinal cancers (expected 9.7), nine of which were cancers of the stomach (expected 6.0). Mortality from lung cancer was excessive for workers with at least 15 years since entry into granite work (latency) (21 observed and 9.5 expected), being highest during the follow-up period of 25-29 years (observed 8, expected 2.1). The results indicate that granite exposure per se may be an etiologic factor in the initiation or promotion of malignant neoplasms. PMID- 3033820 TI - Study on the interaction of Se and erythrocyte membrane--effect of Se on the Na, K-ATPase activity and fluidity of erythrocyte membranes. AB - The content of binding-Se in erythrocyte membrane may affect its structure and function. Decrease in Se content will lead to a lower activity of Na, K-ATPase and lipid fluidity in ghosts, and a marked increase in Na, K-ATPase activity as well as lipid fluidity of erythrocyte membrane was observed by supplementation of low concentration of Na2SeO3 (0.03-0.3 ppm Se), in vitro. This effect was obvious only when the ghosts were incubated with Se at 0-4 degrees C for at least half an hour prior to assay. PMID- 3033819 TI - [Impairment of the immune system caused by drugs]. AB - The immune response and the ensuing inflammation relies on a complex interaction of cells and mediators. Various drugs can interfere with individual steps of the immune response, and in so doing they often imitate regulatory mechanisms of the immune system itself. The immunosuppressive effect of corticosteroids is based on changes in cell migration, reduced responsiveness of monocytes/macrophages to various stimuli and diminished production of interleukin-2. Cyclosporin A appears to block prolactin binding to prolactin receptors on lymphocytes, thus interfering with the immunostimulatory effect of prolactin. It also appears to have a Calmodulin antagonism and might thus block lymphokine production. Anticoagulants may block delayed type hypersensitivity reactions, since activation of the coagulation cascade is involved in this type of immune reaction. Attempts to use calcium channel blockers as immunosuppressive agents, or to take advantage of the immunoregulatory effects of adrenergic substances/blockers or other neurotransmitters, are of experimental value only. PMID- 3033821 TI - Study on the interaction of Se and erythrocyte membrane--protective effect of Se on the erythrocyte membranes. AB - During the ageing of erythrocyte membrane, the spectrin content, Na,K-ATPase activity as well as the lipid fluidity are obviously decreased. However, supplementation of a trace amount of Na2SeO3 in the medium could prevent the dissociation of spectrin from membrane and delay the changes of Na,K-ATPase activity and lipid fluidity. The effectiveness is proportional to Se concentration within the range of 0.1-1.0 ppm. Similar effect of supplementation of Se on the intact erythrocytes during ageing has been also observed. The protective action of Se on biomembranes is generally interpreted in terms of the activity of Se-containing glutathione peroxidase (GSHPx). However, GSHPx mainly distributes in the cytoplasma of erythrocytes, therefore it seems that the protective action of supplemented Se on the isolated erythrocyte membrane might not be related to the activity of GSHPx. PMID- 3033822 TI - HsaI: a restriction enzyme from human being. AB - The HsaI restriction enzyme from the embryos of human, Homo sapiens, has been isolated with both the tissue extract and nuclear extract. It proves to be an unusual enzyme, clearly related functionally to Type II endonuclease. HsaI seems to be an isoschizomer of EcoRI, but it has a distinctive property of elution, differing from EcoRI. Upon SDS-polyacrylamide gel electrophoresis, the enzyme preparation showed Coomassie blue staining bands, having the molecular weight of 65,000 and 22,000 daltons in size, respectively. PMID- 3033823 TI - Clustering of genes dispensable for growth in culture in the S component of the HSV-1 genome. AB - The herpes simplex virus 1 genome consists of one long and one short stretch of unique sequences flanked by inverted repeat sequences. The nucleotide sequence and RNA map predict 12 open reading frames designated as US1 through US12 within the short stretch of unique sequences. This paper reports the construction of virus mutants from which US2, US3, or US4 had been deleted that are capable of growth in cell culture. One of the three deleted genes, US4, specifies the viral envelope glycoprotein G. Mutants with deletions in US1, US8, US9, US10, US11, and US12 have been previously reported. The nine genes deleted from this region form two clusters, US1 through US4 and US8 through US12, and encode at least two and possibly more structural proteins. The presence of so many genes dispensable for growth in cell culture suggests several hypotheses regarding their function and evolution. PMID- 3033824 TI - Rapid identification of nonessential genes of herpes simplex virus type 1 by Tn5 mutagenesis. AB - The large genome of herpes simplex virus type of (HSV-1) encodes at least 80 polypeptides, the majority of which have no recognized function. A subgroup of these gene products appears to be nonessential for virus replication in cell culture, but contributes to the complex life cycle of the virus in the host. To identify such functions, a simple insertional mutagenesis method has been used for selective inactivation of individual HSV-1 genes. The bacterial transposon Tn5 was allowed to insert randomly into cloned restriction fragments representing the entire short unique (US) region of the HSV-1 genome. Of the 12 open reading frames that were mutagenized with Tn5, mutant derivatives of US2, US4, and US5 were recombined into the virus. These three genes proved to be nonessential for HSV-1 replication in Vero (African Green monkey kidney) cells and the US4 gene appeared to be involved in viral pathogenesis in the central nervous system of mice. This rapid mutagenesis procedure should prove useful in exploring the entire HSV-1 genome as well as the genomes of other complex animal viruses. PMID- 3033825 TI - Cloning of large segments of exogenous DNA into yeast by means of artificial chromosome vectors. AB - Fragments of exogenous DNA that range in size up to several hundred kilobase pairs have been cloned into yeast by ligating them to vector sequences that allow their propagation as linear artificial chromosomes. Individual clones of yeast and human DNA that have been analyzed by pulsed-field gel electrophoresis appear to represent faithful replicas of the source DNA. The efficiency with which clones can be generated is high enough to allow the construction of comprehensive libraries from the genomes of higher organisms. By offering a tenfold increase in the size of the DNA molecules that can be cloned into a microbial host, this system addresses a major gap in existing experimental methods for analyzing complex DNA sources. PMID- 3033826 TI - Human T-lymphotropic virus type 4 and the human immunodeficiency virus in West Africa. AB - A new human T-lymphotropic virus (HTLV-4) was recently described in healthy people from Senegal. This virus has many properties in common with members of the human T-lymphotropic viruses, particularly the human immunodeficiency virus or HIV, the etiologic agent of acquired immune deficiency syndrome (AIDS), but does not appear to be associated with immunodeficiency-related disorders. In the present study, serum samples were obtained from 4248 individuals from six West African countries, including Senegal, Guinea, Guinea Bissau, Mauritania, Burkina Faso, and Ivory Coast. These samples, collected during 1985-1987, were from people categorized as healthy control, sexually active risk, and disease populations. All samples were analyzed for reactivity to HTLV-4 and HIV by radioimmunoprecipitation-sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting. Evidence for HTLV-4 infection was found in five of the six countries. The seroprevalence varied markedly from country to country. Healthy sexually active individuals in the risk category had the highest levels of HTLV-4 infection compared to individuals in the healthy control category and the disease category, the latter including AIDS patients. The seroprevalence of HIV infection in most of these countries was quite low, although tightly associated with the rare cases of AIDS. The biology of HTLV-4 infection thus differs from that of HIV in Central Africa or the United States and Europe. The presence of these viruses and their different pathogenicities in several countries of West Africa indicate the necessity for serologic assays that will distinguish between them. Further studies of their origin and distribution as well as of their biology will be important in advancing our understanding of AIDS. PMID- 3033827 TI - Expression of the art/trs protein of HIV and study of its role in viral envelope synthesis. AB - The art/trs transactivator protein of human immunodeficiency virus (HIV) was expressed in mammalian cells as a 19-kilodalton protein that was immunoreactive with sera from HIV-infected patients. Separate plasmids encoding the art/trs protein, the tat protein, or the envelope glycoprotein gp120 were used to demonstrate that both art/trs and tat are absolutely required for the synthesis of gp120 from its cognate messenger RNA. In addition, both the tat and art/trs proteins influence the level of envelope RNA. The results suggest that art/trs and tat may be ideal targets for potential anti-HIV agents in AIDS therapy. PMID- 3033828 TI - Efficient packaging of readthrough RNA in ALV: implications for oncogene transduction. AB - Readthrough viral transcripts are present at relatively high levels in cells infected with avian leukosis virus. It has been proposed that they can function as intermediates in the transduction of proto-oncogenes by retroviruses. It is shown here, by the analysis of viruses containing a mutation in the AAUAAA polyadenylation signal, that readthrough RNAs have the requisite properties to function as transduction intermediates: readthrough RNAs were polyadenylated and packaged as efficiently as normal viral RNA, RNAs nearly 11.2 kilobases (3.5 kilobases larger than wild-type avian leukosis virus genomes) were present in virions of the mutant virus, and virus particles containing both readthrough and normal genomes were most likely infectious. PMID- 3033829 TI - Prosecution urged in fraud case. PMID- 3033831 TI - Meteorological influences on the spread of foot-and-mouth disease. PMID- 3033830 TI - [Effects of phenylephrine on isolated papillary muscles of the guinea pig and rabbit]. PMID- 3033833 TI - Distal duodenal carcinoma and intussusception. AB - Adenocarcinoma of the duodenum is an uncommon malignancy that usually presents itself with either obstructive symptoms or jaundice, depending on its location. The diagnosis should be suspected in any patient with a high small bowel obstruction, weight loss, or chronic abdominal pain. Endoscopy and barium contrast x-ray films are the preferred initial studies; enteroclysis may provide additional information. Surgical resection offers the only hope for long-term survival, which should approach 50% with resectable tumors. PMID- 3033832 TI - Malignant salivary gland tumors of the base of the tongue. AB - We present a 41-year retrospective study of patients with malignant salivary gland tumors of the base of the tongue treated at The University of Texas M. D. Anderson Hospital and Tumor Institute at Houston. This report characterized the patient group, documents their physical findings, analyzes survival, and draws some conclusions regarding clinical course and treatment options. When feasible, surgical resection is the preferred treatment, with planned postoperative radiotherapy when indicated by pathologic findings. PMID- 3033835 TI - [Wilms' tumor in an adult]. PMID- 3033834 TI - The neurovirulence of flaviviruses in crab-eating monkeys (Macaca fascicularis). AB - The neurovirulent properties of attenuated dengue-2 and yellow fever (YF) vaccines, dengue-2 (DEN-2) and Japanese encephalitis (JE) viruses were studied in crab-eating monkeys (Macaca fascicularis). Number of central nervous system sites (as proportion affected) with neurovirulence (NV) lesions were compared. The results indicate that these monkeys reliably developed NV-lesion when inoculated with either JE or YF vaccine viruses (87%). NV-lesions occurred in a minority when inoculated with DEN-2 vaccine virus, were of minimal severity (9%), were probably biologically insignificant, and were of equal or less severity than lesions produced by its parental virus (10%). PMID- 3033836 TI - Anatomic features of the furcal nerve and its clinical significance. AB - Attention must be paid to the furcal nerve when analyzing lumbosacral radicular symptoms, especially when neurologic findings are atypical and the responsible level cannot be assessed. An anatomic and clinical study of the furcal nerve showed the following: the furcal nerve was found in all dissections, and it arises at the L4-root level in most dissections (93%); the furcal nerve has its own anterior and posterior root fibers and its own dorsal nerve root ganglion. This proves that the furcal nerve is an independent nerve root. Neurologic symptoms, suggestive of two roots being involved, are frequently due to furcal nerve compression. PMID- 3033837 TI - Malignant fibrous histiocytoma of the breast. A case report. AB - Primary malignant fibrous histiocytoma of the breast was diagnosed in a 45-year old Indian woman. She presented with a large ulcerated lesion and had lung metastases. Treatment was by radiotherapy, toilet mastectomy and chemotherapy. The patient died of rapid extension of lung metastases 15 months after first being seen. Although this tumour has previously been considered to have a relatively good prognosis, this report emphasises its fully malignant metastatic potential. PMID- 3033838 TI - Dissociation of the aggregating effect and the inhibitory effect upon cyclic adenosine monophosphate accumulation by adrenaline and adenosine diphosphate in human platelets. AB - Recent evidence indicates that adrenaline and adenosine diphosphate each have separate stimulus-response pathways for induction of aggregation and inhibition of cAMP accumulation. We have used a natural model to test the validity of this evidence, i.e. patients from two kindreds with an inherited bleeding disorder due to absent aggregation to adrenaline and no secondary wave response to large concentrations of adenosine diphosphate. Our studies showed that the inhibitory effect of adrenaline and adenosine diphosphate on PGE1-induced increase of cAMP in the patients was not different from that of the controls both for adrenaline and adenosine diphosphate. These results therefore support the present evidence that both adrenaline and adenosine diphosphate apply separate pathways in these two platelet functions. PMID- 3033839 TI - [Muscle relaxation with triethylene thiophosphoramide]. PMID- 3033840 TI - [Infections in pregnant women and newborn infants caused by cytomegaloviruses]. PMID- 3033841 TI - Failure in chymotrypsin enhancement of human rotavirus infectivity. AB - Effects of alpha-chymotrypsin on human rotavirus infectivity and hemagglutination activity were investigated using the KUN strain which was grown in the absence of trypsin and noninfectious. No activating effect of the enzyme on the virus infectivity was found, while HA titers of the virus rapidly decreased after exposure to the enzyme. PMID- 3033842 TI - Assessment of central and peripheral nerve functions in chain-saw operators: a study of short-latency somatosensory evoked potential and peripheral nerve conduction. AB - In order to clarify the effects of local vibration on the peripheral and central nervous system, peripheral (median) nerve conduction velocities and short-latency somatosensory evoked potentials (SSEP) following stimulation of the median nerve at the wrist were measured in 15 male forest workers in 1986. They had engaged in chain-saw operation for 16-34 (mean 22) years; their working days in 1985 averaged 124 days with a range of 50-203 days. The results indicated significant delays in maximal motor and sensory nerve conduction velocities followed by prolongation of all 4 peak latencies of SSEP up to the sensory cortex of the brain (N9, N13, N20 and P23 latencies) in chain-saw operators; their N9 and P23 latencies were significantly correlated with total working days per year. On the other hand, no significant prolongation of the interpeak latencies of SSEP (i.e., cervico-spinobulbar and central conduction times) was found in the workers. It is concluded that local vibration predominantly affects peripheral nerve conduction; cervico-spinobulbar and central nerve conduction may not be significantly affected. PMID- 3033843 TI - Characterized dispersive wave-form of evoked compound muscle action potential in the localized ulnar nerve neuropathy. AB - Disorder of the ulnar nerve with continuity around the elbow is known to be a neuropathy produced by long lasting compression, friction or traction. Forty-four patients with clinical diagnosis of ulnar nerve neuropathy were examined by means of motor nerve conduction study and induced electromyography. Compound muscle action potentials (CMAPs) from hand muscle were evoked by stimulating many points proximal and distal to the elbow and analysis of CMAP was performed. In mild grade of nerve lesion, dispersive wave-form with the extension of its duration was elicited by only stimulating proximal points to the elbow. It was also found that conduction velocity is delayed only over the short distance across the elbow. Thus an initial stage of nerve lesion has a tendency of dysfunctioning of conductivity restricted in only affected portion. CMAP patterns are classified into four groups in comparison with the progress of clinical signs of the disease. This classification can be utilized for the evaluation of the progress of ulnar nerve lesion. PMID- 3033844 TI - The time course of changes in force and cyclic AMP levels produced by isoproterenol and forskolin in isolated rabbit detrusor muscle. AB - The effects of forskolin and isoproterenol on contractile force and cyclic AMP (cAMP) levels were compared in rabbit detrusor. Both forskolin and isoproterenol produced relaxation of rabbit detrusor and an increase in cAMP levels in the tissue. Although the relaxant response to forskolin was similar to that of isoproterenol, the increase in cAMP levels induced by forskolin was significantly larger than that induced by isoproterenol. These results suggest that there is a discrepancy in the relaxation responses and cAMP levels in response to forskolin and isoproterenol. PMID- 3033845 TI - Cyclodiene insecticides inhibit GABAA receptor-regulated chloride transport. AB - The function of GABAA receptors in rat brain was assayed by GABA stimulation of 36Cl-influx into membrane microsacs. The evidence that the measured flux resulted from GABAA receptor activation was supported by the higher potency of muscimol than of GABA in inducing 36Cl-influx with Hill coefficients of 1.71 and 1.87, respectively, suggesting that two agonist molecules were binding to each receptor. Also, the GABA-induced 36Cl-influx was inhibited by the convulsants picrotoxinin and t-butylbicyclophosphorothionate (TBPS), and the competitive inhibitor (+)-bicuculline was more potent than (-)-bicuculline in inhibiting this 36Cl-influx. Furthermore, preincubation of the membranes with pentobarbital or diazepam increased the GABAA receptor's affinity for GABA as judged by the increased 36Cl-influx induced by the same GABA concentration without increasing maximal 36Cl-influx. The GABAA receptor was desensitized as shown by the dose dependent reduction in GABA-induced 36Cl-influx that resulted from preincubation of the membranes with GABA. Seven cyclodienes inhibited the GABA-induced 36Cl influx, with endosulfan I and endrin being more potent than their less toxic isomers endosulfan II and dieldrin, and the epoxides of heptachlor and aldrin (i.e., dieldrin) were more potent than their less toxic parent compounds. There was good correlation (r = 0.9) between inhibition of [35S]TBPS binding to rat brain membranes by these cyclodienes and their inhibition of GABA-induced 36Cl influx. PMID- 3033846 TI - Dose-dependent elevation of Ah receptor binding by TCDD in rat liver. AB - Changes in [3H]TCDD binding to unoccupied liver Ah receptor were examined following chronic or acute administration of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) to adult female Sprague-Dawley rats. Chronic biweekly administration of TCDD equivalent to 0, 10, 30, or 100 ng/kg/day TCDD for 22 weeks caused dose dependent increases in liver TCDD concentration, aryl hydrocarbon hydroxylase (AHH) induction, and [3H]TCDD binding to unoccupied cytosolic Ah receptor sites. Maximal increases (twofold) of cytosolic receptor binding occurred at an estimated dose of slightly greater than 30 ng/kg/day. The increase in [3H]TCDD binding was half-maximal at an estimated dose of approximately 17 ng/kg/day which produced a liver concentration of 1.5 ppb TCDD. Cytosolic [3H]TCDD binding in control and treated animals sedimented mainly in the 8-9 S region of sucrose density gradients with a minor peak sedimenting in the 4-5 S region. Binding was markedly elevated in the 8-9 S region of cytosols from the TCDD-induced rats; however, TCDD treatment had no affect on [3H]TCDD binding in the 4-5 S region. Saturation and Scatchard analyses of Ah receptor binding showed no apparent changes in Kd following chronic TCDD treatment; however, a twofold increase in the number of unoccupied Ah receptor binding sites was observed. Neither aging nor ovariectomy significantly changed measurable cytosolic Ah receptor binding in control animals. When adult female rats were administered a single dose of TCDD (6 micrograms/kg) an initial drop (approximately 40%) in cytosolic receptor binding was observed at 30 and 60 min, followed by a steady increase in binding up to 250% of controls 9 days after TCDD treatment. PMID- 3033847 TI - Conditioned flavor aversions: applications in assessing the efficacy of chelators in the treatment of heavy-metal intoxication. AB - A series of studies investigated the conditioned flavor aversions induced by administration of either lead or thallium in combination with either dimercaprol or dimercaptosuccinic acid in an attempt to correlate changes in flavor-aversion conditioning to changes known to alter the toxicity of metal administration. Rats received po administration of either thallium sulfate or lead acetate given alone or in combination with either dimercaprol or dimercaptosuccinic acid after consuming saccharin. Three days later they were given the choice between consuming saccharin or water, and saccharin preferences were recorded. When compared to rats receiving either nothing or the vehicle, rats receiving either lead or thallium showed significant reductions in saccharin preferences (i.e., conditioned flavor aversions). Rats receiving lead acetate in combination with either of the two chelators displayed significantly reduced conditioned flavor aversions when compared to the aversions induced in rats receiving lead alone. Under the same conditions, there were no differences in the conditioned flavor aversions of rats receiving thallium only and those of rats receiving thallium in combination with either of the two chelators. Attenuation of the lead-induced conditioned flavor aversions was eliminated when chelator administration was delayed by 4 hr. This attenuation of lead-induced but not thallium-induced aversions by dimercaprol and dimercaptosuccinic acid demonstrates the sensitivity and selectivity of the flavor-aversion conditioning paradigm in characterizing metal-chelator interactions and is in agreement with clinical reports of effective chelation therapy in cases of lead but not thallium intoxication. PMID- 3033848 TI - Factors affecting the response of the female rat reproductive system to cannabinoids. AB - Chronic oral administration of either crude marihuana extract (CME) or delta 9 tetrahydrocannabinol (THC) to female Fischer rats for 64-72 days, at a dose approximating heavy usage by humans, reduces food intake by about 8%. Pair feeding studies demonstrate that this decreased food intake accounts for previously described decreases in uterine and ovarian weights, which are much more affected by food restriction than is body weight. THC-treated rats lost weight initially which was not regained. Pair-fed rats gained only about one-half of the weight of the untreated control or vehicle-treated control rats over a 64 day period. Although long-term cannabinoid administration leads to tolerance and the resumption of the estrous cycle, the onset of estrus is often delayed when cannabinoid is administered 5-6 hr before the proestrus luteinizing hormone (LH) surge. Our results indicate that although chronic exposure to cannabinoids can continue to affect the rat estrous cycle, they do not have a direct effect on growth of the reproductive organs. The results reemphasize the need for adequate nutritional controls in marihuana and other toxicological research. PMID- 3033849 TI - Polybrominated dibenzo-p-dioxins and related compounds: quantitative in vivo and in vitro structure-activity relationships. AB - The effects of structure on the in vitro receptor binding affinities, aryl hydrocarbon hydroxylase (AHH) and ethoxyresorufin O-deethylase (EROD) induction potencies in rat hepatoma cells were determined for the following compounds: 2 bromo-, 2,7/2,8-dibromo-, 2,3,7-tribromo-, 2,4,6,8/1,3,7,9-tetrabromo-, 2,3,7,8 tetrabromo-, 1,3,7,8-tetrabromo-, 1,2,3,7,8-pentabromo-, 1,2,4,7,8-pentabromo-, 2,3-dibromo-7,8-dichloro-, 2,8-dibromo-3,7-dichloro- and 2-bromo-3,7,8 trichlorodibenzo-p-dioxin. The structure-activity relationships (SARs) for the polybrominated dibenzo-p-dioxins (PBDDs) were comparable for both in vitro responses: the most active compounds were substituted only in the lateral 2,3,7 and 8 position and the addition of non-lateral or removal of lateral halogen substituents reduced the activity of the resultant compound. The biologic and toxic effects of 2,3,7,8-tetrabromo-, 1,3,7,8-tetrabromo-, 1,2,4,7,8-pentabromo 1,2,3,7,8-pentabromo-, 2-bromo-3,7,8-trichloro- and 2,3-dibromo-7,8 dichlorodibenzo-p-dioxin on several receptor-mediated responses (thymic atrophy, body weight loss, hepatic microsomal AHH and EROD induction) were determined in a dose-response fashion in immature male Wistar rats. A comparison of the ED50 values for the in vivo responses demonstrated that the SARs for the PBDDs and brominated polychlorinated dibenzo-p-dioxins were comparable to those observed for in vitro receptor binding and AHH induction. Moreover, there was an excellent linear correlation between the -log EC50 (in vitro AHH induction) vs. the in vivo -log ED50 (thymic atrophy) and -log ED50 (body wt loss) correlation coefficient, r = 0.97 for all 2 correlations). PMID- 3033850 TI - [Parotid gland function following radioiodine therapy of thyroid cancer based on data from radionuclide sialoscintigraphy]. PMID- 3033851 TI - Receptor radioligand system for measuring digoxin activity. AB - The purpose of this study was to develop and evaluate a receptor radioligand assay (RRA) for the measurement of digoxin, its metabolites, and endoxin and to compare the results of this assay with those of a fluorescence polarization immunoassay (FPIA) for digoxin. The within-run coefficients of variations for the RRA at digoxin concentrations of 2.0 and 5.0 nmol/L were 21 and 10%, respectively. Serum samples collected from 33 volunteers (not on digoxin medication) measured in the RRA resulted in levels less than 1.23 nmol/L digoxin equivalents (1.23 nmol/L is the lower level of sensitivity for this system). Serum samples collected from 29 patients receiving digoxin for therapeutic purposes and measured in the RRA gave a mean of 3.39 nmol/L digoxin equivalents. The samples collected from the patients receiving digoxin were also measured using FPIA with a mean of 1.75 nmol/L digoxin. The interference of progesterone, 11-alpha-hydroxyprogesterone, cortisone, and six digoxin metabolites on the binding of 3H-ouabain in the RRA was also evaluated. PMID- 3033852 TI - De novo hepatocellular carcinoma without chronic liver disease but with 17 years of azathioprine immunosuppression. PMID- 3033853 TI - The effect of prednisolone, thromboxane, and platelet-activating factor receptor antagonists on lymphocyte and platelet migration in experimental cardiac transplantation. AB - Three agents that significantly prolong cardiac allograft survival were tested in Lewis rats that were recipients of hearts from Lewis X Brown-Norway F1 hybrid donors. In the presence of azathioprine, the effects of daily administration of either the thromboxane antagonist (L 640,035), the platelet-activating factor (PAF) antagonist (BN 52021) or prednisolone were evaluated on the infiltration of cardiac allografts by syngeneic lymphocytes and platelets labeled with 111indium. As anticipated, platelet deposition was reduced by the thromboxane antagonist and unaffected by the PAF antagonist; the latter is likely due to the known absence of PAF receptors in rat platelets. In addition prednisolone had no effect. The increased accumulation of lymphocytes on days 4-5 was also unaffected by all three drugs. These experiments indicate that, in this model, graft survival is not necessarily related to lymphocyte and platelet infiltration of the graft. The data also provide evidence for the efficacy of the thromboxane receptor antagonist L 640,035 in preventing platelet deposition in vivo. PMID- 3033854 TI - Association of rheumatoid factors in renal transplant recipients with cytomegalovirus infection and not with rejection. AB - Because rheumatoid factors (RF) were detected in the circulation of the majority of early renal transplant recipients and could be eluted from rejected transplants, RF were hypothesized to be related to antibody responses to the histoincompatible graft. The possibility that RF production might have been related to infection and not rejection has not been considered previously. Therefore, we investigated serial serum samples from 147 adult renal transplant recipients for RF with latex agglutination and radioimmune assays. RF were detected in the sera of 32 patients, 30 of whom had coincident active cytomegalovirus (CMV) infections. Another 45 patients with active CMV infections did not have detectable circulating RF. In contrast, of 74 patients who experienced a total of 103 treated reversible or irreversible rejection episodes in the absence of evidence of active CMV infections, only 2 patients produced RF during their rejection episodes. Nine of the patients who did not produce RF during a rejection episode subsequently produced RF during a later CMV infection. These data indicate that RF production in renal transplant recipients is associated with CMV infection and not rejection. Moreover, RF production was found to be more frequently associated with primary and severe CMV infections than with secondary or milder CMV infections. RF production was not more frequent in patients who were HLA-DR-4-positive., older, or female, characteristics that have been associated with RF production in other populations. All of the sera with detectable RF contained IgM antibodies that were directed to the Fc portion of human IgG, and about half contained additional IgM antibodies directed to Fab. Thus CMV infections may be the stimuli for the IgM anti-Fab antibodies that have been reported in pretransplant serum samples. Eleven patients produced IgG or IgA RF in addition to IgM RF during CMV infections. PMID- 3033856 TI - Systemic evaluation of Sjogren-like syndrome after bone marrow transplantation in man. AB - A systematic evaluation of Sjogren-like syndrome (SLS) was performed in 68 bone marrow transplant (BMT) recipients (60 allogeneic and 8 syngeneic recipients). At day 100, the patients underwent clinical evaluation, functional salivary scintigraphy, and lip biopsy. If any findings were abnormal, the examinations were repeated annually for 3 years. Twenty-two patients with SLS and extensive chronic graft-versus-host disease (CGVHD) had abnormal scintiscan and lip biopsy at day 100. Marked keratoconjunctivitis sicca and xerostomia developed between 12 and 24 months after BMT and, thereafter, progressively decreased. Twenty-seven irradiated recipients (7 syngeneic and 20 allogeneic recipients without CGVHD) had isolated xerostomia and disturbed scintiscan but normal biopsy. Seven other patients with limited CGVHD had a lymphocytic infiltrate on lip biopsy but no SLS and a normal scintiscan. Schirmer's test, functional salivary scintigraphy, and lip biopsy allowed us to distinguish SLS from radiotherapy sequelae. As early as day 100, these 3 tests have a predictive value for SLS, one of the criteria for extensive CGVHD. PMID- 3033855 TI - T lymphocyte subpopulations in bone-marrow-transplanted patients in relation to graft-versus-host disease and cytomegalovirus-induced infection. AB - T cell subpopulations were studied in peripheral blood from 62 bone marrow recipients using monoclonal antibodies and flow cytometry. Patients with acute graft-versus-host disease (GVHD), patients with chronic GVHD, and patients with cytomegalovirus (CMV) infection had decreased OKT4/8 ratios. The data suggest that in GVHD this was due to reduced absolute numbers of OKT4-positive cells. CMV infected patients had significantly increased numbers of OKT8-positive cells. Monitoring of T cell subpopulations was not informative for diagnosis or prognosis of acute GVHD in the single patient. CMV infection should be suspected at a sudden increase in the number of OKT8-positive cells. PMID- 3033857 TI - Reproducible high yield of rat islets by stationary in vitro digestion following pancreatic ductal or portal venous collagenase injection. AB - Pancreatic distension with collagenase solution followed by stationary in vitro digestion yields large numbers of intact islets. We compared in rats two routes of collagenase injection, pancreatic ductal (PD) and portal venous (PV), for islet yield, in vitro insulin secretory capacities, and in vivo functional viability. The islet yield in the PD method (n = 11) was greater than that in the PV method (n = 8) (682 +/- 27 vs. 417 +/- 39 per pancreas, P less than 0.025). The insulin release from the PD islets in response to 16.7 mM glucose increased gradually following culture, 3.2 +/- 0.8 ng/10 islets/30 min (fresh) to 12.3 +/- 2.1 (24-hr culture). In contrast, insulin release from the PV islets increased during the first 6 hr of culture, but decreased after 24 hr in culture. Under electronmicroscopic examination, the PD islets revealed a well preserved structure with healthy endocrine cells, while the PV islets showed a dilated capillary network and distorted endocrine cell continuity. When 100 PD islets were transplanted into streptozotocin-induced diabetic B6AF1 mice (n = 8), all the recipient mice restored normoglycemia (less than 200 mg/dl) within 1-4 days following transplantation and maintained it until rejection. However, the recipient mice given 100 PV islets showed a significant delay in restoring normoglycemia, and 3 of 8 mice given 100 PV islets were still hyperglycemic on day 4 postgrafting. In summary, pancreatic ductal collagenase injection followed by stationary in vitro digestion reproducibly yields higher numbers of intact and viable islets when compared with portal venous collagenase injection, indicating the superiority of this method to portal venous injection. PMID- 3033858 TI - A light microscopic and ultrastructural study of spindle-shaped hepatocellular carcinoma. AB - Spindle-cell hepatocellular carcinoma is an unusual morphologic variant of hepatocellular carcinoma with a typical sarcomatous appearance. The exact diagnosis of this tumor may be difficult when only small biopsies are available and in the absence of ultrastructural studies. We describe two cases of hepatocellular carcinoma: one was entirely composed of spindle-shaped cells, and the other was a typical hepatocellular carcinoma with only a small area of sarcomatous, fusiform cells. In the first case, ultrastructural studies demonstrated desmosomes and many Mallory bodies and confirmed the epithelial nature of the neoplasm. In the second case, no ultrastructural studies were available, but the presence of gradual transition from liver cell carcinoma to spindle-cell carcinoma excluded the diagnosis of carcino-sarcoma. PMID- 3033859 TI - Spontaneous massive necrosis of a hepatocellular carcinoma. AB - A case of hepatocellular carcinoma that underwent total necrosis without previous chemotherapy is described. Histologic examination of the neoplasm revealed massive thrombosis of numerous peritumoral venous vessels in the adjacent normal liver. Although the importance of a newly formed arterial blood supply for the maintenance of the viability of hepatocellular carcinoma is unquestionable, this case suggests a similar importance of the venous drainage of the surrounding liver. PMID- 3033860 TI - [The role of vasopressin and oxytocin in the regulation of glycemia levels and carbohydrate metabolism in the liver]. AB - Vasopressin and oxytocin administered subcutaneously and intravenously in a dose of 0.5 IU/kg were studied in experiments on albino male rats for their effect on the glycogen content and gluconeogenesis enzymes activity in the liver as well as on the glycemia level. Neurohormones injected subcutaneously have no effect on the values of the measured indices. Vasopressin already the first 15-60 min after its intravenous injection in the mentioned dose leads to an essential decrease of the glucose content in blood, glycogen amount, glucose-6-phosphatase and fructose 1.6-diphosphatase (EC 3.1.3.9 and 3.1.3.11) activity in the liver of test animals. The intravenous injection of oxytocin in the same dose induces changes in the carbohydrate metabolism indices similar in their direction and magnitude to the effects of intravenous injection of vasopressin. PMID- 3033861 TI - [Phosphorylase and gluco-6-phosphatase activity and glycogen level in tissues of vertebrates]. AB - The activity of phosphorylase (EC 2.4.1.1), glucose-6-phosphatase (EC 3.1.3.9) and the content of glycogen have been determined in tissues of fish, amphibians, reptiles, mammalians. No differences in the activity of phosphorylase and glucose 6-phosphatase in the liver, myocardium, and brain of animals of the phylogenetic groups under study are found. The activity of glucose-6-phosphatase in the anaerobic muscles of poikilothermal animals is found to be rather high. The share of phosphorylase a in the skeletal muscles and brain as well as the glycogen content in the brain of these animals is essentially higher than that of adult mammalians. PMID- 3033862 TI - [Uptake of [14C]GABA by rat brain slices; the effect of Ca2+]. AB - The [14C]GABA uptake by slices (0.3 mm thick) of Wistar rat brain cortex was studied for its dependence on the GABA concentration in the medium, time of incubation and the presence of Ca2+. This process is characterized by the absence of saturation; the uptake by slices increases sharply when the concentration of exogenous [14C]GABA reaches 200 microM. Bicucullin (10(-4) M), an antagonist of GABA, inhibits the accumulation of GABA in the concentration of 0.2 microM by 60%, that evidences for a considerable contribution of the receptor binding to this process. The [14C]GABA uptake when Ca2+ is absent in the incubation medium and when its concentration is 10(-3) M is practically the same and comparatively low concentrations of Ca2+ (10(-6)-10(-4] decrease the GABA uptake. PMID- 3033863 TI - [Radiographic examination of the ileum reservoir (S-pouch) and anal anastomosis]. PMID- 3033864 TI - [Alternative treatment of children--why and how often? A questionnaire study at a pediatric department]. PMID- 3033865 TI - [Adrenocortical function in geriatric patients evaluated before and after administration of ascorbic acid]. PMID- 3033866 TI - Signet-ring cell carcinoma of bladder. Evaluation of three cases with review of literature. AB - Three cases of adenocarcinoma involving the bladder and consisting mainly of signet-ring-shaped cells are presented. Each case represents a different form that the lesion may take, and these forms are compared. This unusual neoplasm has a worse prognosis than transitional cell carcinoma of the bladder, possibly because of factors that lead to delays in management. Radiation therapy is ineffective, and the results of treatment with segmental resection of the involved area of the bladder are equivalent to those of total cystectomy. PMID- 3033867 TI - Obturator neuropathy after multiple genitourinary procedures. AB - Management of neuropathic complications of surgery associated with multiple genitourinary procedures can prove difficult. A case of obturator neuropathy is reported. A rational approach for diagnosis and management is presented as well as possible mechanisms for its presence. PMID- 3033868 TI - [The effect of stress on blood levels of vitamin A and E in piglets]. AB - In five trials with 99 suckling or weaned piglets, the effects of the increased adrenocortical function, caused by cold, weaning from the sow or fasting or by stimulation with exogenous adrenocorticotrophic hormone (ACTH) were studied as exerted on vitamin A and vitamin E concentrations in the blood serum. Three hours after an exposure of three-day and four-day piglets to the temperature of 8-12 degrees C, a small drop of the concentrations of both vitamins occurred. In four week weanling piglets a decrease in vitamin E concentration was observed in two days, the trend being slight in vitamin A concentration. At the same time some sibs were left fasting, which considerably reduced the concentrations of both vitamins. The situation was similar in two hours after ACTH administration to suckling piglets, however the difference was insignificant in vitamin E concentrations. In seventeen hours elapsing from two administrations of the adrenocorticotrophic hormone (ACTH) when the increased secretion of corticosteroids was fading out, only the vitamin A concentration in suckling piglets was found to drop. The response in weanling piglets was negligible. The suppressive effects of stress on vitamin A concentrations were usually observed when the levels of circulating corticosteroids were high or in the period immediately following this status. The changes in nutrition after early weaning exert large negative effects on vitamin E concentrations in the blood serum. The differences in the response of the organism to the two vitamins may be due to various types of transport mechanisms in the blood circulation. The specific effects of stress factors are mentioned. PMID- 3033870 TI - Effect of vaccination of the dam on rotavirus infection in young calves. AB - Vaccination of cows with a combined, inactivated, adjuvanted rotavirus and Escherichia coli vaccine resulted in increased neutralising antibody titres to rotavirus in serum and colostral whey. Evidence was obtained that vaccination resulted in a decreased incidence of rotavirus shedding and of abnormal faeces or diarrhoea in young calves fed colostrum and milk from the vaccinated dams. The E coli component of the vaccine was not evaluated because no natural challenge was evident. PMID- 3033869 TI - Hereditary bovine syndactyly: diagnosis in bovine fetuses. AB - Diagnostic guidelines were established for progeny testing of hereditary bovine syndactyly. Through the use of superovulation and embryo transfer, 139 fetuses were recovered at 50 to 77 days gestation. Normal (+/+, +/sy) and syndactylous (sy/sy) anatomy of Holstein fetuses was defined, and the accuracy of macroscopic versus microscopic limb diagnosis was assessed. Chondrification and ossification differences between normal (+/+, +/sy) and syndactylous (sy/sy) fetuses were only age-related. Normal (+/+, +/sy) fetal limbs differed from normal (+/+, +/sy) adult bovine limbs in two ways. Fetal metacarpal and metatarsal III and IV bones were not fused, and fetal metacarpal and metatarsal II and V bones often extended up to three-fourths the length of metacarpal and metatarsal III and IV bones. In syndactylous (sy/sy) fetuses, synostosis asymmetries occurred within and between fetal limbs, and between fetuses, representing variable gene expressivity. Synostosis pattern within limbs did not correspond with those of the adult bovine; the second phalangeal pair was synostotic most frequently in the fetus, followed by the first, and then the third pair. Synostosis patterns between fetal limbs agreed with those of the adult; there was a right-left and front-rear limb gradient. Partial synostoses occurred sporadically in all three paired phalanges. Those of the first and third pair always involved the tip closest to the second phalangeal pair. A unique example of variable gene expressivity occurred in one syndactylous fetus. Both front limbs were syndactylous, while both rear limbs were normal grossly. Microscopically the right rear limb was normal while the left rear limb consisted of closely apposed phalangeal blastemata without coalescence.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3033871 TI - Effect of vitamin D supplementation during pregnancy on the vitamin D status of ewes and their lambs. AB - Pregnant ewes were injected intramuscularly with 300,000 iu of vitamin D3 in a water miscible vehicle either 10, seven or four weeks before the expected lambing date and the effects on plasma concentrations of 25-hydroxyvitamin D3 were monitored. The concentrations increased quickly and remained high at parturition but at no time were they outside the normal physiological range. The concentrations in the plasma of the newborn lambs were higher than in uninjected controls and were well correlated with the concentrations in their mothers. Dosing pregnant ewes with 300,000 iu of vitamin D3 in a rapidly available form, approximately two months before lambing, provided a safe means of increasing the vitamin D status of the ewe and the newborn lamb by preventing the seasonally low concentrations of 25-hydroxyvitamin D3. PMID- 3033872 TI - Incidence, characterisation and prophylaxis of nephropathogenic avian infectious bronchitis viruses. PMID- 3033873 TI - Maedi-visna virus infection in commercial flocks of sheep in East Anglia. PMID- 3033874 TI - Desorption of porcine parvovirus from aluminum hydroxide adjuvant with subsequent viral immunoassay or hemagglutination assay. AB - Cell culture fluids containing porcine parvovirus were adjuvanted with varying concentrations of aluminum hydroxide gel. Adsorption of virus and total protein to adjuvant was proportional to adjuvant concentration. Desorption of virus and protein from the adjuvant in substantial, reproducible quantities was achieved by washing adjuvanted preparations with 1.2 M potassium phosphate, followed by dialysis and concentration of wash fluids. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of unadjuvanted viral fluids, supernatants of adjuvanted mixtures, and desorptive washings of the corresponding adjuvant pellets revealed no qualitative differences in protein banding patterns. Desorbed virus was quantitated by a hemagglutination assay, or by an enzyme-linked immunoassay employing a monoclonal anti-porcine parvovirus antibody. Virus desorption and subsequent assays permitted in vitro estimation of virus content in adjuvanted porcine parvovirus preparations. This approach may be useful in estimating the antigen content of inactivated aluminum hydroxide adjuvanted veterinary vaccines and reducing the extent of required in vivo testing. PMID- 3033875 TI - Morphological clues for the diagnosis of small hepatocellular carcinomas. AB - Histological features of 44 cases of small hepatocellular carcinoma (HCC) were examined and compared with those of large regenerative nodules. The highly differentiated type of HCC most often occurred in nodules which were less than 2 cm in diameter. Noticeably, in 9 out of 15 such cases (60.0%), tumour cells were arranged in trabeculae of almost normal thickness (normotrabecular pattern). These trabeculae, however, showed variable nuclear crowding, occasional microacinar formation, and increase in cytoplasmic basophilia. It is emphasized that the presence of this triad may be a very reliable indicator for the histological identification of early HCC, especially in examining limited material such as a biopsy specimen. However, cellular and structural atypia becomes more prominent in nodules which are larger than 2 cm. PMID- 3033877 TI - [Possible relation between viruses and oromaxillofacial tumors. I. Demonstration of herpes antigens and anti-herpes antibodies]. AB - Herpes virus type 1 and 2 antigens were detected in cells originating from malignant tumors with oromaxillofacial localization. A higher incidence of type 1 antigens was noted, as compared to the type 2. Investigated antigens were detected more frequently in the tumors of ectodermal origin. Antiherpesvirus FC antibodies were found in 74.5% of the tumor bearing subjects, but only in 22.3% of the controls. PMID- 3033876 TI - Virus expression. EGF and transferrin receptors in human papillomas. AB - Thirty five non regressing cutaneous and mucosal human papillomas were studied for the expression of EGF and transferrin receptors by indirect immunofluorescence on frozen sections. The lesions were also examined for the presence of human papillomavirus (HPV) DNA by in situ-hybridization with biotinylated probes and viral capsid antigen. The mapping of EGF and transferrin receptors was modified in cutaneous lesions with drastic viral cytopathic effects and was enhanced in mucosal lesions mainly in laryngeal papillomas, which are poor virus producers. The greatest increase in EGF and transferrin receptor reactivity was observed in the group of mucosal lesions in which viral DNA was more frequently detected than viral antigen. This suggests that viral DNA may play a role in basal cell stimulation. Moreover some of these lesions with dense inflammatory reactions showed DR antigen expression by epithelial cells. Our findings indicate that epithelial cell activation in papillomas might be modulated by other factors than HPV such as mediators of the local immune response. PMID- 3033878 TI - [Possible relation between viruses and oromaxillofacial tumors. II. Research on the presence of the SV40 antigen and specific antibodies in patients with oromaxillofacial tumors]. AB - SV40 antigen was detected on sections through malignant oromaxillofacial tumors in 47.8% of the cases, with a higher incidence among the epithelium originating tumors. Anti-SV40 CF antibodies were detected in 83.6% of the examined subjects with cancer, but only in 57.9% of the blood donors. The titers were quite low in general. The results are discussed from the point of view of the immunologic status of the patients. PMID- 3033879 TI - [Possible relation between viruses and oromaxillofacial tumors. III. Demonstration of the SV40 antigen and anti-SV40 antibodies in patients with tumors of the parotid gland]. AB - SV40 antigen was detected in 7 of 13 malignant tumors developed in the head and neck region. Specific complement fixing antibodies were found in all the patients with the SV40 antigen present in the parotid gland tumoral cells. Incidence of the anti-SV40 complement fixing antibodies in parotid gland tumor bearing patients was of 69.6%. PMID- 3033880 TI - [Possible relation between viruses and oromaxillofacial tumors. IV. Presence of the herpes antigen and anti-herpes antibodies in patients with tumors of the parotid gland]. AB - The presence of herpesvirus antigen and of antiherpesvirus antibodies was detected in 46% and in 36.3%, respectively, of the patients bearing parotid gland tumors. Very high titers of antiherpesvirus antibodies were found in 70.6% of the patients with nasopharyngeal carcinoma. PMID- 3033881 TI - [Possible relation between viruses and oromaxillofacial tumors. V. Demonstration of hemagglutination-inhibiting anti-BK virus antibodies in patients with tumors of the parotid gland]. AB - Anti-BK-virus hemagglutination inhibiting antibodies were revealed in 81.8% of the patients with parotid gland tumors. Results of the investigations conducted on oromaxillofacial tumors including the parotid gland ones are discussed from the point of view of the presence of viral antigens (herpes-, SV40 and BK viruses) and of specific antibodies. Possible implication of the papova viruses in the etiopathogenesis of the parotid gland tumors in humans are also discussed. PMID- 3033882 TI - Sindbis virus mutants resistant to mycophenolic acid and ribavirin. AB - Previous work from this laboratory has demonstrated a correlation between the inhibition by ribavirin (Rbv), mycophenolic acid (MPA), or 2-amino thiadiazole (TDA) of Sindbis virus replication in Aedes albopictus mosquito cells and a reduction in cellular GTP levels. This reduction in GTP results from the inhibition by these drugs of inosine monophosphate dehydrogenase (IMPDH), the first enzyme specific for the de novo synthesis of GMP. By serial passage of SV in A. albopictus cells in the presence of 25 microM MPA, we have now isolated viral mutants which are highly resistant not only to MPA but also to Rbv and TDA. For example, whereas 500 microM Rbv reduced the plaquing efficiency of SVSTD by at least 10(6)-fold, the same concentration of Rbv reduced the plaquing efficiency of the MPA-resistant mutants less than 5-fold. This is the first example of a viral mutant resistant to the antiviral compound Rbv. PMID- 3033883 TI - Further analysis of the role of calcium in rotavirus morphogenesis. AB - Previously we reported that calcium plays an important role in the maturation of bovine rotavirus (M. S. Shahrabadi and P. W. K. Lee, 1986. Virology 152, 298 307). We now demonstrate that the formation of mature double-shelled (L) particles was strictly dependent on the concentration of calcium present in the growth medium. The formation of single-shelled (D) particles did not appear to be a calcium-mediated process. Subsequent labeling studies using 45Ca revealed that calcium was incorporated into the L particles but not the D particles. The previously noted decreased level of the outer capsid protein VP7 (42K) in calcium deprived cultures was now found to be due to the preferential degradation, and not to the impaired synthesis, of this protein in the absence of calcium. It was further demonstrated that calcium had a stabilizing effect on VP7 and that VP7 synthesized in the presence of calcium was not degraded upon subsequent calcium deprivation. Protein degradation during calcium deprivation was apparently limited to the mature form of VP7 since the unglycosylated precursor (pVP7), formed in the presence of tunicamycin, was found to be stable under this condition. Electron microscopic examination of infected cells revealed that in the presence of calcium, virus maturation took place by the budding of viral cores through the endoplasmic reticulum (ER). No such budding was observed in calcium-deprived cells. In these cells mature virions were absent and membrane fragments could be found associated with viral cores or single-shelled particles. PMID- 3033885 TI - Variability of pseudorabies virus glycoprotein I expression. AB - The 130,000 mol wt glycoprotein I (gI) derived from two approx 80-kDa precursors is one of the major constituents of the envelope of pseudorabies virus (PRV) strain Phylaxia. Recently, gI has been shown to be nonessential for PRV replication since several PRV vaccine strains with deletions in the region of the genome encoding the gI gene have been described. In this paper we demonstrate that other alterations affecting gI expression can occur. We describe a PRV field isolate which expresses a single gI precursor molecule pgI of 64,000 mol wt. This precursor is processed into 60,000 mol wt gI. In contrast to PRV Phylaxia, the gI expressing isolate is not neutralized by anti-gI monoclonal antibodies. Virions expressing the pgI also emerged after serial in vitro passages of the wild-type PRV strain NIA-5 which initially expressed wild-type pgI. Concomitant with the appearance of pgI the pgI disappeared and the resistance of the virus population to neutralization by anti-gI monoclonal antibodies increased. Furthermore, the amount of expression of gI and pgI in single plaque isolates of the PRV strain Ka was found to be highly variable among different plaque isolates and correlated with a different susceptibility to neutralization by anti-gI monoclonal antibodies. In single plaque isolates of strain Phylaxia, however, gI expression appeared to be stable. In all cases, no genomic or transcriptional alterations could be observed. Thus, viruses resistant to anti-gI antibodies occur spontaneously in vivo and in vitro, which argues against the use of gI as a subunit vaccine. PMID- 3033886 TI - Growth of malignant rabbit fibroma virus in lymphoid cells. AB - To understand better the immunosuppressive capacity of malignant rabbit fibroma virus (MV), we characterized MV growth in lymphoid cells. Replication of MV occurs in unstimulated normal spleen cells in vitro and is enhanced by adding T- or B-lymphocyte mitogens. In splenic T-lymphocyte preparations, comparable results are found: virus growth in the absence of mitogen, augmented by adding Con A. Unlike mature T cells, thymic lymphocytes support MV replication only when mitogen is added. When spleen cells from rabbits infected with MV in vivo are removed and cultured without mitogen, MV growth is again observed, with virus titer increasing about 10-fold per day of culture. In spleen cell populations from MV tumor-bearing rabbits, MV grows best in T lymphocytes, moderately in B lymphocytes, and least efficiently in adherent cells. When spleen cells are examined immediately following sacrifice, MV antigens are expressed solely on T lymphocytes from rabbits infected in vivo with MV 7 days previously. However, following overnight incubation in vitro a population of non-T lymphocytes displays cell membrane virus antigens. MV adapts itself somewhat to growth in lymphocytes, showing significantly greater growth in lymphocytes following passage in lymphocytes than is observed for non-lymphocyte-propagated virus. MV infected lymphocytes also elaborate a factor that enhances MV growth in lymphocytes. Thus, MV replicates preferentially in mature T lymphocytes but will grow well in B cells as well. In vivo infection produces relatively small amounts of recoverable virus. However, when these lymphocytes are cultured in vitro virus replicates very well without added mitogens. These growth patterns may help to understand MV-induced immunologic dysfunction. PMID- 3033884 TI - Fate and expression of simian virus 40 DNA after introduction into murine cells under nonselective conditions. AB - When SV40 infects mouse cells, it does not replicate but instead causes neoplastic transformation of a small percentage of the cells. It is unknown, however, what happens to the virus in those cells that do not become transformed. We introduced SV40 into mouse cells by nonselective means, either by cotransfection of SV40 DNA with a selectable marker or by random cloning of SV40 infected cells. We analyzed the fate of viral DNA sequences, expression of T antigens, and transformation properties of these cells. We found that, upon infection, viral DNA integration occurs at a frequency that is at least 10-fold higher than the frequency of transformation. The majority of these cells are not transformed due to lack of expression of T antigen. One cell line which expresses a truncated T antigen is not transformed. We have mapped the viral sequences in the genome of these cells and find that integration in the large T intron is probably responsible for the defect. Lack of transformation can therefore be attributed to both cellular and viral factors, namely, introduction of viral DNA into cells that are resistant to transformation or integration of viral DNA in such a way that T antigen expression is prohibited. PMID- 3033887 TI - Differences in virus-induced polypeptides in cells infected by cytopathic and noncytopathic biotypes of bovine virus diarrhea-mucosal disease virus. AB - Two biotypes of bovine viral diarrhea-mucosal disease virus are present in nature: one that induces cytopathology in infected bovine cells and the other that infects cells without overt cytopathology. Infections with both types of virus yield similar amounts of infectious progeny virus. Field and laboratory isolates of both biotypes of bovine viral diarrhea (BVD) virus were analyzed by radioimmunoprecipitation and polyacrylamide gel electrophoresis of infected cell extracts. The noncytopathic biotype BVD (NCB-BVD) virus isolates can be differentiated from cytopathic biotype BVD (CB-BVD) isolates on the basis of peculiar polypeptide profiles they induce in the infected cell. The most abundant polypeptide in CB-BVD infected cells is the 80K polypeptide. NCB-BVD virus infected cells lack the 80K polypeptide and induce a predominant 118K polypeptide. D-[2-3H]Mannose labeling of cells infected with NCB-BVD indicated that at least three polypeptides are N-glycosylated: 75K, 56K-58K, and 48K. In addition the sizes and ratios of the glycoproteins induced by all virus isolates showed a marked variation. We present evidence indicating that there is remarkable heterogeneity among the field viral isolates of BVD and this methodology is of potential value for molecular epidemiology studies. PMID- 3033888 TI - Analysis of the early regulatory region of the human papovavirus BK. AB - BKV is a human papovavirus which latently infects a majority of the world population and whose DNA has been found in human tumor tissue. Along with simian virus 40 (SV40) and JCV, it is one of several highly homologous polyomaviruses which display distinct host ranges, tissue tropisms, and transformation potentials. Determination of these properties is thought to reside, in part, in the noncoding regulatory region of these viruses. We have studied the regulation of gene expression by the early promoter and enhancer of BKV. Our results show that the early promoter of BKV consists of elements found both to the early side of and within the proximal 18 bp of the first enhancer element itself. At least one BKV regulatory element appears to be downstream of the mRNA start sites. The BKV enhancer consists of two different types of elements, three direct repeats, and an element (denoted "c") found in the 30 bp to the late side of the distal repeat. When used with the BKV promoter the enhancer repeat elements were found to be redundant, optimal promoter activity requiring only two of the three repeats plus the c element. Using the heterologous SV40 promoter the optimal BKV enhancer consisted of either three repeats or two repeats plus the c element. Subfragments of the enhancer region were capable of partial activation of homologous and heterologous promoters. We conclude that the regulation of BKV early gene expression involves novel elements arranged in ways not previously described in other papovaviruses. PMID- 3033890 TI - Isolation and preliminary characterization of temperature-sensitive mutants of human cytomegalovirus. AB - Thirteen temperature-sensitive (ts) mutants of human cytomegalovirus (HCMV) have been isolated after mutagenesis with nitrosoguanidine or ultraviolet light. Four mutants defective in viral DNA synthesis at nonpermissive temperature (DNA-) were classified into two separate complementation groups. The other nine HCMV ts mutants, capable of synthesizing viral DNA at the elevated temperature (DNA+), belonged to seven independent complementation groups. The number of essential genes encoded by HCMV is thus raised here by nine. PMID- 3033889 TI - Altered structure and expression of c-myc in feline T-cell tumours. AB - The c-myc gene is rearranged in a subset of feline T-cell lymphosarcomas. Detailed mapping of c-myc rearrangements showed that some result from feline leukaemia virus (FeLV) proviral integration within or upstream of c-myc, but one case involves a complex 3' alteration and amplification which is apparently not directly virus-induced. S1 nuclease mapping of RNA from normal cells using c-myc probes revealed two presumptive 5' ends, each corresponding to a promoter-like sequence (P1 and P2), and a major 3' discontinuity which mapped to the 3'-most of two possible polyadenylation signals. Analysis of RNA from a series of tumours revealed different modes of c-myc expression. All tumours produced P1 and P2 transcripts with apparently normal structure except for one case where an insertion in intron 1 displaced exon 1 sequences. The abundance ratio of P1/P2 transcripts varied considerably and was high in tumours which carry a rearrangement adjacent to c-myc, but some other T-cell tumours with no apparent myc alteration displayed an equally high ratio. However, a consistent feature was the lack of detectable RNA from normal c-myc alleles in tumours which express a rearranged c-myc allele or a transduced FeLV v-myc gene. We suggest that this may prove to be a useful indicator of the presence of an oncogenically active myc gene, whether this is a rearranged c-myc or transduced v-myc sequence. PMID- 3033891 TI - Molecular cloning and nucleotide sequence analysis of several naturally occurring HPV-5 deletion mutant genomes. AB - Three deletion mutants of naturally occurring human papillomavirus type 5 (HPV-5) were molecularly cloned into phage vectors. The nature of these deletions was characterized initially by restriction endonuclease mapping and electron microscopic heteroduplex analysis and ultimately by nucleotide sequence analysis. The sizes of the deletions are 353, 1329, 1571, and 2267 bp and map to the late gene region of the HPV-5 genome. The 80 nucleotides immediately adjacent to the deletions exhibit no significant detectable sequence homologies or symmetries and therefore were probably not formed by the sequence-dependent events of homologous recombination or site-specific recombination. PMID- 3033892 TI - Nucleotide sequence and genome organization of human papillomavirus type 5. AB - Human papillomavirus (HPV) type 5 is associated with benign and malignant lesions of the disease epidermodysplasia verruciformis (EV). Because of the strong correlation between the presence of HPV-5 and malignant progression in these patients, we have elucidated the nucleotide sequence of the HPV-5 genome. The size of the HPV-5 genome is 7746 nucleotides and its organization is similar to that of other papillomaviruses. The HPV-5 genome exhibits extensive sequence homology with another EV-associated papillomavirus, HPV-8, although HPV-5 appears to contain at least one additional open reading frame. PMID- 3033893 TI - Mechanism of entry of human rhinovirus 2 into HeLa cells. AB - Internalized human rhinovirus 2 (HRV2) undergoes a rapid conformational change leading to recognition by the C-determinant-specific monoclonal antibody 2G2. In the presence of the ionophore monensin, the virus accumulates in the cells in its native conformation and infection is strongly inhibited. At 20 degrees but not at 34 degrees the inhibitory effect of monensin can be overcome by a short incubation of the infected cells at low pH as late as 2 hr after inoculation. Incubation of infected cells at 20 degrees prior to addition of monensin permits virus synthesis to occur, depending on the time of preincubation. PMID- 3033894 TI - Effects of inhibitors of the cytoplasmic structures and functions on the early phase of infection of cultured cells with simian virus 40. AB - To obtain information about cytoplasmic structures and functions involving the entry of simian virus 40 virions into cells, we examined whether the inhibitors that affect the functions and/or structure of lysosomes, cell membrane, and cytoskeletons inhibit expression of nuclear T antigen in the SV40-inoculated rat 3Y1 and monkey CV-1 cells. Chloroquine, methylamine, and butylamine did not inhibit T-antigen expression, suggesting that lysosomal acidification is not required for establishment of infection. Cytochalasin B had no effect, suggesting that microfilaments are not involved. Monensin, colcemid, and amantadine each inhibited T-antigen expression at doses causing no obvious cytotoxicity. Maximal inhibition was seen when these inhibitors were added to the cultures within 1 hr (monensin), within 4 hr (colcemid), or within 12 hr (amantadine) after virion adsorption to the cell surface. When the inhibitor was present in the virus inoculated cultures for 24 hr and then removed, nuclear T antigen began to be expressed at 4 hr (monensin), 9 hr (colcemid), or 1 hr (amantadine) after removal of the inhibitors. Results of SDS-PAGE analysis of immunoprecipitated radiolabeled proteins of infected cells revealed that amantadine inhibited synthesis of large and small T antigens as well as general protein synthesis. Inhibition by colcemid may be due to disruption of microtubules, because other microtubule-disrupting agents (colchicine, vinblastine, nocodazole, and podophyllotoxin) also inhibited appearance of nuclear T antigen but lumicolchicine and taxol did not. Electron microscopy revealed that, in the presence of colcemid, although the adsorbed virions were readily internalized to form pinosomes, vectorial movement of the pinosomes to the nucleus appeared to be inhibited. Results of electron microscopy also suggest that inhibition by monensin may occur mainly in internalization of adsorbed virions and that the inhibition is leaky such that the early steps of infection proceed slowly in the presence of monensin. We conclude that monensin, colcemid, and amantadine interfere with mutually different early events of SV40 infection. PMID- 3033895 TI - The creation of adenovirus genomes with viable, stable, internal redundancies centered about the E2b region. AB - During the course of constructing new adenoviral strains by overlap recombination, we have discovered that internally redundant viable genomes can be created by end-to-end joining of the input DNA molecules. The cellular functions responsible for the end-joining activity frequently ligated the overhanging single strands of the complementary ends to form a novel restriction site at the junction. In 2 of the 17 cases analyzed in detail by restriction digestion, and some sequence determinations, the cellular functions had repaired the ends, presumably prior to end-joining. Four of the isolates had suffered deletions at the junction ranging in size from 13 to 532 bp. The isolate with the largest deletion also had an insertion of 14 bp of unknown origin at the site of the deletion. All of the redundant isolates replicated as efficiently as isogenic unit length strains, and plaque dilution titrations obeyed one-hit kinetics, showing that the redundant genomes were nondefective. Nevertheless unit-length genomes were observed at a low level (some 5 to 10% of the total) in stocks of each isolate before and after plaque purification. They presumably arose by recombination between the redundant sequences either intra- or intermolecularly. Evidence from Southern blot analysis showed that molecules with three copies of the redundant sequences also arose and could be detected both in intracellular and in capsid viral DNA. These species would arise by unequal crossing-over between redundant genomes. The efficient replication of the redundant species demonstrates that the precise spatial relationships between splice donors and acceptors on either strand, in this region of the genome, do not have to be rigidly maintained. These data suggest that it may be possible to place other genetic information between the DNA polymerase and terminal protein precursor genes and have it expressed from the major late promoter in its normal location. PMID- 3033897 TI - Nucleotide sequence of a radiation leukemia virus genome. AB - The complete nucleotide sequence of an infectious molecular clone of a radiation murine leukemia proviral DNA RadLV/VL3(T+L+) has been determined. The sequence of the RNA genome is 8318 nucleotides long and contains three large open reading frames encoding the gag, pol, and env gene products. With the exception of a xenotropiclike R peptide and the LTR which bears structural similarities to a xenotropic LTR, displaying typical enhancerlike sequences, the remaining sequences are strikingly similar to the endogenous, ecotropic Akv murine leukemia virus. Therefore, it could be postulated that the leukemogenic properties of RadLV/VL3(T+L+) were generated by a recombination event between a xenotropic virus and an Akv-like ecotropic virus. PMID- 3033896 TI - Regulation of equine herpesvirus type 1 gene expression: characterization of immediate early, early, and late transcription. AB - The regulation of equine herpesvirus type 1 (EHV-1) transcription was examined in infected rabbit kidney cells using metabolic inhibitors. In order to map EHV-1 immediate early, early, and late transcripts, viral RNA was 32P-labeled in vivo and hybridized to EHV-1 DNA restriction fragments immobilized on nitrocellulose filters. Immediate early viral RNA was mapped to one region of the viral genome within the inverted repeat DNA sequences (map units 0.78-0.83 and 0.95-1.0). Northern blot hybridization analysis using a 32P-labeled cloned DNA probe from this region identified a single immediate early viral transcript (approximately 6 kb). Transcription of early and late genes was not restricted to any specific region on the viral genome as indicated by the ability of 32P-labeled early and late RNA to hybridize to EHV-1 restriction endonuclease fragments from both the long and short components of EHV-1 DNA. Additional experiments performed without the use of metabolic inhibitors confirmed that EHV-1 transcription is temporally regulated. The characterization of EHV-1 transcription during productive infection will serve as a reference for the analysis of viral transcripts in oncogenically transformed and persistently infected cells. PMID- 3033898 TI - [Effect of hepatectomy on liver adenylate cyclase activity in rats of various ages]. AB - Age-dependent differences were found in patterns of rat liver adenylate cyclase system in response to hepatectomy-induced proliferation. After hepatectomy a decrease in basic activity of adenylate cyclase and in content of cAMP were observed only in adult animals. At the same time, the epinephrine-stimulated enzymatic activity was decreased in old hepatectomized rats as compared with intact animals. These alterations in regulatory activity of the adenylate cyclase system appear to affect the intensity of DNA biosynthesis and the functional activity of regenerating liver tissue of old rats. PMID- 3033900 TI - [Leukotrienes--a new class of physiologically active compounds (review of the literature)]. PMID- 3033899 TI - [Effect of thymectomy on activity of key enzymes of gluconeogenesis in the rat liver]. AB - Activities of key enzymes of gluconeogenesis--phosphoenolpyruvate carboxykinase, fructose-1,6-diphosphatase, glucose-6-phosphatase as well as the content of glycogen were studied in liver tissue of thymectomized rats. Gluconeogenesis was inhibited in liver tissue of these rats. As thymus and adrenal cortex are firmly related, the inhibition of gluconeogenesis in liver tissue of thymectomized rats appears to occur due to a decrease in the glucocorticoid activating effect followed the inhibition of adrenal cortex functions and absence of thymus hormones directly responsible for carbohydrate metabolism. PMID- 3033901 TI - [Antiviral activity of the 1-alpha- and beta-desoxy-D-ribofuranosides of 5 trimethylsilyl uracil]. PMID- 3033902 TI - [Use of immunoenzyme analysis for the diagnosis of rotavirus gastroenteritis in children]. PMID- 3033903 TI - [Rotaviruses and rotavirus gastroenteritis]. PMID- 3033904 TI - [Alphavirus replication: the cascade proteolytic shearing of virus-specific polyproteins and virion morphogenesis]. PMID- 3033905 TI - [Expression and mutagenesis of genes coding for protein synthesis in the influenza virus]. AB - Double-stranded cDNA copies of the neuraminidase genes of influenza viruses A/Tokyo/3/67 (N2), A/tern/Australia/G70C/75 (N9), and B/Lee/40, and the hemagglutinin genes of A/Memphis/1/71 (H3) and B/Hong Kong/8/73 were cloned into a SV40 vector in which the late region was replaced by the influenza sequences. Thus the influenza genes were expressed in transfected cells under the control of the SV40 late promoter. The Tokyo/67 neuraminidase gene was modified by oligonucleotide-directed site-specific in vitro mutagenesis. Several of the amino acid residues which are conserved in all known neuraminidases and which line the sialic acid binding pocket were changed, and the mutant gene ligated back into the SV40 vector. Five mutations which have been fully characterized resulted in synthesis of a protein which had totally lost neuraminidase enzyme activity. PMID- 3033906 TI - [Effect of gamma radiation on the components of hepatitis B virus and the delta antigen]. AB - The influence of ionizing irradiation on the serological activity of purified HBsAg, receptors to polymerized albumin, DNA-polymerase activity, delta agent, and delta antigen was studied. Different radiosensitivity of hepatitis B virus components and delta-system was demonstrated. The possibility of sterilization by irradiation of purified HBsAg preparations suitable for construction of a vaccine against hepatitis B was shown. PMID- 3033907 TI - [Suppression of rotavirus SA-11 reproduction by protease inhibitors in cell culture]. AB - The effect of proteases inhibitors, epsilon-amino-caproic acid and gordox, on reproduction of rotavirus SA-11 in MA-104 cells was studied by enzyme immunoassay. These inhibitors were shown to exert an inhibiting effect on rotavirus reproduction. PMID- 3033908 TI - [Isolation and cultivation of a swine rotavirus]. AB - A swine rotavirus capable of inducing the cytopathic effect was isolated in a roller culture of Macaca rhesus kidney cells (line MA-104) after two preliminary passages in gnotobiotic piglets and colostrum-free piglets, and the isolate was designated strain K. For virus isolation, fecal specimens were treated with trypsin, and besides, trypsin was added into the maintenance medium. After 20 passages in MA-104 cell culture the swine rotavirus was adapted to pig embryo kidney cell cultures (SPEV line) in which the maximum virus accumulation, 8.0 log TCD50/ml, was achieved within 24 hours after inoculation. The virus accumulation was most marked in the presence of 10 micrograms/ml trypsin in the maintenance medium. In the roller culture, the virus multiplied to a much higher titre (approximately 100-fold) than in the stationary culture. In the course of passages the virus was shown to lose its pathogenic properties. A scheme of swine rotavirus virion structure is suggested on the basis of ultramicroscopic studies. PMID- 3033909 TI - [Electrophoretic analysis of the proteins of different alphavirus strains]. AB - Proteins of various alphavirus strains: Venezuelan equine encephalomyelitis, Semliki Forest, and Sindbis, were studied by a high resolution polyacrylamide gel electrophoresis. In addition to structural C, E1 and E2 proteins, the infected cells were found to contain a number of nonstructural polypeptides: B, 83 kD (analogue of nsP2), 75 kD (nsP3), PE2, precursor of E2 and a product of modification of nucleocapsid protein C27. In virions, in addition to the main structural polypeptides, protein components with molecular weights about 100 kD were found which, most likely, were aggregates of E1/E2 glycoproteins, and 40 kD, a product of E1 protein degradation. The alphavirus strains under study differed both in the electrophoretic mobility of the above-mentioned virus-specific polypeptides and in the stability of B and PE2 proteins in the infected cells. The intracellular stability of polyprotein B depended considerably also on the host cell. PMID- 3033910 TI - [Experimental coronavirus encephalomyelitis in mice]. AB - A combined comparative virological, morphological, and immunological study of experimental coronavirus encephalomyelitis was carried out in mice in order to elucidate the pathogenetic mechanisms involved in the formation of the foci of lesions in acute and chronic forms of the disease. Intracerebral inoculation of C3H mice with the neurotropic JHM strain of murine hepatitis virus induces a disease with demyelinization foci in the CNS running acute, subacute, or chronic course. This model underlies a concept that demyelinating diseases are caused by viruses producing immunopathologic responses realized via certain histocompatibility loci. The long-term persistence of viral antigen in the CNS and liver may be explained by virus replication in hepatocytes and oligodendrocytes at low levels, and exacerbations of the disease are prevented by the immune system. PMID- 3033911 TI - AIDS-related neuropathy. PMID- 3033912 TI - [Freeze-etching study of the replication of the infectious bovine rhinotracheitis virus]. PMID- 3033913 TI - [A case of carcinoid tumor of the bronchus with an unusual course diagnosed and treated as small cell carcinoma of the lung]. PMID- 3033914 TI - [Insulin-secreting pancreatic islet cell tumor (insulinoma) treated with diltiazem and surgery]. PMID- 3033915 TI - [Role of enkephalins in the functioning of the bladder and urethra]. PMID- 3033916 TI - [Diagnostic difficulties in a case of laryngeal nonchromaffin paraganglioma]. PMID- 3033918 TI - Endocrine abnormalities in human temporal lobe epilepsy. AB - Patients with temporal lobe epilepsy secrete ACTH at higher rates and in greater amounts than normal subjects. Temporal lobectomy restores ACTH secretion to normal amounts and rates. The ACTH secretion in temporal lobe epilepsy is independent of anticonvulsant drug effect and seizure frequency. Electrical stimulation of medial temporal lobe structures in patients with temporal lobe epilepsy affected ACTH secretion in a manner consistent with the hypothesis that ACTH secretion is regulated by tonic inhibition. A defect in the excitatory and/or inhibitory components of this regulatory process appears to exist in temporal lobe epilepsy. PMID- 3033917 TI - Nutritional aspects of osteoporosis. PMID- 3033920 TI - [Staging of inoperable small cell bronchial cancer]. AB - This paper compares two staging regimes for the inoperable small-cell carcinoma of the lung. It reveals that at present the differentiation in two classes of the Veterans Administration Lung Cancer Study Group (VALCSG) is preferred to the TNM classification of the World Health Organization (WHO), as it includes an orientation of prognosis and enables consequences of therapy relevant to practice. If studies of therapy of the small-cell carcinoma of the lung are compared, the uniform application of the classification should be considered. PMID- 3033919 TI - [Recent knowledge about the significance of 1,25-hydroxyvitamin D and the development of genetically-induced rickets]. AB - Receptors for the binding of 1,25-hydroxyvitamin D are present in many cell types and are of importance for the regulation of the transcription and function. The 1,25-hydroxyvitamin-D-receptor complexes are in the cells under physiological conditions mainly in the nucleus. The 1,25-hydroxyvitamin D already in a very low concentration in the bone cells further the replication of the DNA and the division of the cells, in the lactotrophic cells of the anterior pituitary the secretion of prolactin, in the B-cells the secretion of insulin and in the epidermis the formation of keratin. In the parathyroid gland it restricts the formation of preproparathormone and in the cultures of lymphocytes the division of cells evoked by the addition of mitogens. The multiplication of various forms of tumour cells and their metastatic spread is inhibited by the administration of 1,25-hydroxyvitamin D3. The equipment with receptors for the 1,25-hydroxyvitamin D is regulated. Biochemical aspects of the development of the 3 different forms of genetically conditioned rachitis are shown. PMID- 3033921 TI - [Current clinical aspects of changes in the lung parenchyma caused by the interaction of polychemotherapy and radiotherapy in the treatment of small cell bronchial cancer]. AB - The use of radiotherapy in the treatment of bronchial carcinoma evokes changes of the healthy lung parenchyma which may lead to pneumonitis and fibrosis. At present more than 20 tumour chemotherapeutics with lung-toxical effect are known, which may induce a pneumonitis or a fibrosis. When two or more lung-toxically acting cytostatic drugs are combined, we have to take into consideration interactions with synergistic character on the healthy lung parenchyma. The combination of a lung-toxical cytostatic drug with the radiotherapy or the combination of a polychemotherapy with the radiation may cause pneumonitides and fibrosis with increased mortality rate. In our therapy programmes for the treatment of the small cell bronchial carcinoma the polychemotherapy of the tumours (I. cyclophosphamide, methotrexate, nitrosomethylurea, II. doxorubicin (adriamycin), dacarbacin, vincristine, III. cyclophosphamide, adriamycin, methotrexate) and the radiotherapy were simultaneously used. Hereby a pneumonitis was x-ray-diagnostically ascertained in more than 70% and a fibrosis in 30-80% of the patients treated. In our opinion these high rates of changes of the lung parenchyma have their cause in a complex process: interaction of the tumour chemotherapeutic drugs among one another; interaction of the tumour polychemotherapy with the radiotherapy (particularly in simultaneous application); the bronchial carcinoma is, depending upon anatomical localisation of the tumour and size a risk factor for an infection; the immunosuppressive situation of the patient increases the danger of an infection particularly in the area of the respiratory tract during the therapy. The course of a pneumonitis is decisively determined by a secondary infection. PMID- 3033922 TI - [Surface analysis of Bio-Sinter ceramic specimens using Rutherford backscattering spectrometry following subcutaneous implantation]. AB - The formation of a calcium and phosphorous containing surface-layer is one of the essential suppositions of bioactive materials for the replace of bone. In the submitted study on sintered Bio-Vitro-ceramic specimens, which where explanted from the subcutaneous tissues of rats after 20, 40, 90 and 180 days, a relative increase of calcium content, and an absolute increase of phosphorous content of the direct surface of the specimens (approximately 0.01 micron) could be demonstrated. In contrast to not implanted specimens the values of silicat were 90% lower. After 180 days of implantation the distribution of elements at the surface of all types of sintered specimens had been stabilized 1.6 (Ap40KS15), 7.1 (Ap40KS30), 2.3 (Ap40SV). PMID- 3033923 TI - [Prenatal diagnosis and obstetric management of a case of Klippel-Trenaunay-Weber syndrome]. AB - A description is given of a case of Klippel-Trenaunay-Weber syndrome diagnosed by sonography in the 28th week of pregnancy. The ultrasound findings established in the course of pregnancy are presented. Whereas minor variants of this syndrome normally do not represent a birth hindrance, a cesarean section was indicated in the present case on the basis of the size of the deformation--especially at the hip--an in order to save the child. PMID- 3033924 TI - [Measuring coronary circulation using an inert gas method--a comparison of common indicator gases]. AB - Validation studies of inert gas techniques are limited in number and usually have not included circumstances with marked heterogeneity of flow. This study was intended to investigate the validity of the method in experimental animals under various hemodynamic conditions by parallel application of helium, argon, krypton and xenon as indicators and by comparison with direct flow measurements. Gases were applied by single breath inhalation. In order to avoid the limitations of conventional methods arising from the systemic recirculation of tracer and from shortened measuring periods we used continuous mass spectrometric recording and numeric deconvolution of the dilution curves in arterial and coronary venous blood. When the transit times are evaluated by "stochastic" analysis and commonly used tissue/blood partition-coefficients (lambda He = 0.95, lambda Ar = 1.1) the helium and argon values are found in reasonable agreement with reference flows. The effects of a shortening of measurements that are based on single integrated samples (Argon method) are shown under control conditions, adenosin infusion and elevated ventricular filling pressure. With xenon (lambda Xe = 0.7) coronary blood flow above 100 ml/min X 100 g is underestimated by the "stochastic" and by the "initial slope technique"; parallel measurements with argon suggest that in the initial slope technique a lambda Xe between 0.8 and 0.9 might be more adequate. PMID- 3033925 TI - [Simultaneous staining of the isoenzymes of adenylate kinase, adenosine deaminase and 6-phosphogluconate dehydrogenase]. PMID- 3033926 TI - Inhibition of cAMP-phosphodiesterase by molybdate. AB - Inhibition of 3':5'-cyclic-AMP-5'-nucleotidohydrolase (EC 3.1.4.17) type II (cAMP phosphodiesterase) by sodium molybdate was studied: While determination of inorganic phosphate and 5'-ribonucleotide phosphohydrolase (5'-nucleotidase) (EC 3.1.3.5) activity was not disturbed by sodium molybdate in concentrations up to 10 mM, cAMP-phosphodiesterase was inhibited by millimolar concentrations of molybdate. The half maximal effect was observed at about 2 mM sodium molybdate (0.75 mM cAMP in the assay). PMID- 3033927 TI - Role of bovine mammillitis virus towards preparation of an alternative vaccine against herpes simplex virus infections of human subjects. AB - Bovine mammillitis virus (BMV) cross-reacted in neutralization and radioimmune assay with herpes simplex virus (HSV) and pre-immunization with BMV protected against challenge by type 2 HSV. There was no evidence to suggest a pathogenic role for BMV as adjudged by a literature search or field enquiry and BMV specific antibody was not detected in 21 human sera or in four sera from personnel engaged in research with BMV; in addition there was no replication or antigen synthesis by BMV in explants of human tissue or cell lines of human origin. It is proposed that BMV might provide an alternative vaccine against HSV infections of human subjects. PMID- 3033928 TI - Correlation of 140S antigen dose with the serum neutralizing antibody response and the level of protection induced in cattle by foot-and-mouth disease vaccines. AB - An analysis was made of data from potency tests on fifteen batches of monovalent foot-and-mouth disease vaccine, comprising five batches each of type O, type A and type C. Regressions were calculated for the relation of percentage protection (probit) versus log 140S antigen dose and for the serum neutralizing antibody titre (log SN50) versus log 140S antigen dose. Type O vaccines required a far higher level (220 ng) of 140S antigen to achieve a 50% protection level (PA50) in cattle than did type A (2.4 ng) and type C (4.36 ng) vaccines. Type O antigen, dose for dose, was as effective at provoking neutralizing antibody as the types A and C antigens. Thus, it would appear that a far higher log SN50 value (2.14) was required for type O vaccines to equate with 50% protection of cattle than was required for type A (1.17) and type C (1.41) vaccines. Prior to 1977, however, the PA50 value for type O vaccine strain was only 1.34 and it was concluded that an antigenic shift was the most likely cause for the large difference between that value and the current PA50 value. PMID- 3033929 TI - [Study of human phagocytizing cells in clinical medicine]. PMID- 3033930 TI - Stimulating effect of heat shock on the early stage of human cytomegalovirus replication cycle. AB - The effect of mild heat shock on the replication of human cytomegalovirus (HCMV) was studied in human embryo fibroblasts. Treatment of cell cultures at 44 degrees C for 10 min just before infection or at 24 h post infection (p.i.) shortened HCMV eclipse period and enhanced viral replication, while heat shock performed at 48 h p.i. had no effect on the HCMV replication cycle. Study of HCMV-induced early and late antigens confirmed that the cellular response to heat shock influences HCMV replication in the early stage of the viral replication cycle. PMID- 3033931 TI - Vero cells persistently infected with Tacaribe virus: role of interfering particles in the establishment of the infection. AB - Eight Vero cell sublines (Vero T) persistently infected with wild type Tacaribe virus replicated in different hosts were established. In order to unravel the mechanism involved in the initiation and maintenance of persistence, the properties of virus shed by the sublines and the presence of interfering particles (IP) were analyzed. During the course of infection, persistent virus (Tac-pi) underwent mutations although no consistent pattern of virus evolution was observed. ts mutants were isolated from two Vero T sublines, whereas a slow growth variant was shed by another. The remaining sublines released virus resembling wt parental virus. Except for Vero T1 sublines, Vero T cultures shed no detectable IP. These results emphasize the point that neither the emergence of virus mutants nor the synthesis of IP is essential for the maintenance of the persistent state. To define the role of IP in the initiation of persistence, coinfection experiments with a characterized inoculum were performed. For that purpose, attempts were made to obtain IP stocks free from pfu by serial transfers of undiluted virus. Neither enrichment nor amplification of IP occurred, and virus stocks were freed of infectious virus by UV irradiation. If normal Vero cells were infected with Tac-pi virus released by Vero T2, Vero T3, Vero T4, Vero T5, Vero T6, Vero T7 and Vero T10 sublines, a complete destruction of the monolayer without cell recovery was observed. In contrast, parental and Vero T1 viruses always originated persistently infected sublines. Similarly, the addition of IP to virus inocula constituted by Tac-pi viruses released by Vero T2, Vero T3, Vero T4, Vero T5, Vero T6, Vero T7 and Vero T10 sublines gave rise to persistently infected cultures. These results suggest that although IP are not important by themselves in the maintenance of persistence, they play a major role in initiation. PMID- 3033932 TI - Herpes simplex virus gene products involved in the induction of chromosomal aberrations. AB - The effect of short-term herpes simplex virus type 1 (HSV-1) infection on chromosomes of human diploid fibroblasts was examined. In addition to chromosomal breaks, gaps and pulverization, three kinds of cytogenetic damage (double minutes, polyploidy and endoreduplication) not yet reported following productive infection with HSV or other animal viruses were frequently observed. Consistent with previous studies suggesting that the expression of immediate-early and/or early viral gene products is required for the induction of chromosomal damage, was the observation that cells infected at the nonpermissive temperature with HSV 1 temperature-sensitive mutants defective in the gene for the immediate-early transcriptional regulatory protein, ICP4, and three early viral gene products- DNA polymerase (pol), the major HSV DNA-binding protein (ICP8) and an HSV-2 mutant defective in alkaline nuclease--exhibited altered patterns of chromosomal damage relative to the effects of wild-type virus on infected cells. These findings suggest a direct or indirect role for all four gene products in the induction of chromosomal damage. In cells infected with wild-type virus for 4 h or longer, HSV proved to be a more potent mitotic arresting agent than colcemid. Moreover, studies with selected mutants indicate that HSV pol specifically may be involved in mitotic arrest. Additionally, in cells infected at the non-permissive temperature with a pol mutant, the number of polyploid metaphases was reduced 4 fold relative to that seen in wild-type virus-infected cells suggesting a role for HSV pol in the amplification of cellular DNA. PMID- 3033933 TI - MHV nucleocapsid synthesis in the presence of cycloheximide and accumulation of negative strand MHV RNA. AB - We have found that genomic RNA synthesis is inhibited by cycloheximide in cells infected with mouse hepatitis virus, strain A59 (MHV-A59), in agreement with previously published results (Sawicki, S.G. and Sawicki, D.L. (1986) J. Virol, 57, 328-334). In the present study, the fate of the residual genomic RNA synthesized in the presence of cycloheximide was determined. Nearly all of the genomic RNA synthesized in the presence of drug was incorporated into nucleocapsid structures, suggesting that even in the absence of protein synthesis, genomic RNA synthesis and encapsidation are coupled in MHV-infected cells. Sufficient free nucleocapsid N protein was available for this purpose, since the pool of soluble N protein was determined to decay with a half-life of approximately one hour. Negative strand RNA is the template for the synthesis of both genomic and subgenomic positive strand RNA, and would be predicted to accumulate primarily during the early phases of the lytic cycle. In agreement with this prediction, negative strand RNA accumulated during the first 5-6 h of infection, with little additional accumulation occurring over the next 2.5 h. In marked contrast, positive strand RNA increased 5-6-fold over the same 2.5 h period. These results, taken in conjunction with published data, suggest that negative strand RNA is synthesized during the early period of the infectious cycle and is stable in infected cells and also suggest that treatment with cycloheximide at late times does not inhibit positive strand RNA synthesis indirectly by blocking the formation of negative strand templates. PMID- 3033934 TI - Mutational analysis of DNA sequences affecting the replication of defective polyomavirus variant D-50. AB - The genome of the defective polyomavirus variant D-50 consists of tandemly repeated DNA segments. The repeat unit corresponds to a 17% fragment of polyomavirus DNA (Griffin and Fried (1975) Nature 256, pp. 175-179). To allow mutational analysis, a monomer unit of D-50 DNA was cloned. After excision from the plasmid and ligation to form a random mixture of products, circular head to tail oligomers of the cloned segment were replicated in transfected cells. Those molecules had the same replication properties as original D-50 DNA. Deletion of base sequences assumed to be at the origin of DNA synthesis inhibited the replication completely, whereas a deletion of a segment at the junction of the tandem repeats had only a slight inhibitory effect. Mutation of potential coding sequences of the variant genome had a slight stimulatory effect on DNA synthesis, ruling out that D-50 expresses any protein that stimulates replication. In an attempt to construct variants similar to D-50, cells were transfected with polyomavirus DNA fragments including the sequences of the D-50 monomer unit. However, all these molecules replicated very slowly, suggesting the presence of an element that inhibited DNA synthesis. The combined data show that D-50 genomes can be reconstituted by ligation of monomer units and that the origin of DNA replication was the only essential element of the variant genome, whereas other elements had cis-acting auxiliary functions. PMID- 3033935 TI - [Role of cAMP in providing for the plastic properties of the electro-excitable membrane of neurons]. AB - In isolated snail brain, the role was studied of cyclic adenosine monophosphate (cAMP) in providing plastic properties of electro-excitable neuronal membranes of two types, habituating and non-habituating to rhythmic intracellular stimulation with depolarizing electric pulses. It has been shown that at high level of cAMP in the cell maintained with administration of dibutyryl-cAMP and (or) blockaders of phosphodiesterase in incubation medium, habituating cells lose their ability of habituation to stimulation. There is also no habituation in the presence of serotonin: serotonin effect is removed by imidazol, activator of phosphodiesterase. Imidazol promotes the development of habituation of cells, initially non-habituating to stimulation. Data are obtained on connection of Ca2+ effects and cAMP metabolism in habituating cells. On the basis of the obtained data it is suggested that the cyclase system controls plastic properties of neurones of both types, and reduction of cAMP content in the cell apparently mediates the above mentioned Ca-K-mechanism of habituation. PMID- 3033937 TI - [Effect of serotonin and noradrenaline on the magnitude of the response of the command neurons for defensive behavior in Helix lucorum L]. AB - Changes in synaptic responses of identified command neurones of avoidance behaviour to the electric nerve stimulation were investigated in the isolated nervous system of the snail during bath application of serotonin or noradrenaline. Serotonin (10(-5) M) elicited an increase of summary EPSP amplitude in the cells without changes of input resistance and resting potential. Noradrenaline (10(-5) M) application evoked an increase of EPSP amplitude, accompanied by an increase of the input resistance. Mechanisms of serotonin and noradrenaline influence on synaptic responses are discussed. PMID- 3033936 TI - [Reflection of the plastic properties of monosynaptically interconnected neurons in the statistical characteristics of their spike activity]. AB - By mathematical and biomathematical methods of neuronal interaction modelling, changes were studied of cross-correlation histograms (CCH) of impulse flows and of average interimpulse intervals of monosynaptically interconnected neurones, at changes of efficiency of forward and backward connections, of excitability of neurones and of summate action on them of independent random afferent synaptic inflows. It is shown, that a single sign of efficiency increase of monosynaptic excitatory or inhibitory connection between neurones (amplitude increase of the corresponding postsynaptic potential) consists in amplitude increase of the main peak or trough the rated CCH of their impulse flows, followed by a decrease of average interspike intervals of both neurones. PMID- 3033938 TI - [Orientation-exploratory reaction of mice with different genotypes after injection of the ACTH4-10 fragment]. PMID- 3033940 TI - Current trends in leukocyte activation. PMID- 3033939 TI - [Synaptic response of the command neuron for defensive behavior of Helix lucorum L. during exposure to a constant magnetic field]. PMID- 3033941 TI - Relationship between leukemia virus genomes and histocompatibility genes. PMID- 3033942 TI - Pathogenesis of inflammatory bowel disease. PMID- 3033943 TI - Modulation of interferon-gamma induction of Ia expression. Implications for mechanisms of interferon-gamma signal transduction. PMID- 3033944 TI - Lymphocyte hybridomas: monoclonal antibodies and more. PMID- 3033946 TI - Natural killer cells and tumor immunity: 1985. PMID- 3033945 TI - Organization and expression of murine T cell receptor genes. PMID- 3033947 TI - The role of T cell immunity in infection with the herpes group viruses. PMID- 3033948 TI - Immunologic developments in AIDS--1985. PMID- 3033949 TI - [Histologic classification of breast cancer and its precancerous stages]. AB - A proliferating mastopathy with severe epithelial atypia as well as with so called radial scars, intraductal papillomatosis, and findings of so-called lobular cancerisation are regarded as possible precursors of mammary carcinoma. The histological differential diagnosis can be difficult between atypical epithelial proliferations in mastopathy and noninvasive intraductal carcinoma, between tubular formations in radial scars and tubular carcinoma, papilloma and papillary carcinoma as well as between primary and secondary lobular cancerisation. Noninvasive and invasive breast carcinomas can be diagnosed according to the WHO classification (12) with a relatively good reproduceability. Besides the histological type, other macroscopic (tumour size, number of metastatically involved lymph nodes, distant metastases), microscopic (tumour grading) as well as biochemical findings (receptor status) are also important for treatment and prognosis of breast carcinoma. PMID- 3033950 TI - [Pathogenesis, clinical aspects, morphology, therapy and prognosis of phyllodes tumors of the breast]. AB - A case is reported of a malignant cystosarcoma phylloides. After 7-year-follow-up period an increase in size was observed in a mammary nodule, first suspected diagnostic ascertainment. Histology of the removed tumor with an unsuspicious tissue sheath revealed a malignant cystosarcoma. Consequently, any palpable breast node suspected to be fibroadenoma should be removed with short-spaced follow-up examination or at least observed over long periods of time in view of the possible malignant degeneration despite of absent criteria of malignancy. PMID- 3033951 TI - [Estimating the biological effect of detrimental substances on E. coli with flow microcalorimetry. II. Studies using 3 antibiotics]. AB - The influence of three antibiotics (Penicillin G, Oxtetracycline, Polymyxin B), which show different modes of action against gram-negative bacteria, is tested on the growth of liquid E. coli cultures in a flow microcalorimeter. The antibiotics were added during the logarithmic growth phase of the culture when a heat flux of 40 mu W/ml was approached. A supply vessel and a reaction vessel were used for establishing of the baseline and for performing the test, respectively. When testing Oxytetracycline and Polymyxin B the germ content in the nutrient broth was determined, additionally. Some measurements of the optical density of the nutrient broth were carried out parallel to the microcalorimetric investigation on the effect of Polymyxin B. As was to be expected, Penicillin G shows an influence on E. coli at concentrations of about 50 microgram/ml medium, only. An influence of Oxytetracycline on the heat production can be observed at concentrations between 0.3 and 0.6 microgram/ml. Polymyxin B acts on the power time-curve at 0.025 microgram/ml. The minimum doses of Oxytetracycline and Polymyxin B found by the microcalorimetric technique are distinctly lower than those found by bacterial count and by measuring the optical density. Microcalorimetric results are achieved within 2 hours after adding the test substance to the bacterial culture. Flow microcalorimetry seems to be a promising tool for testing antibacterial drugs. PMID- 3033952 TI - The effect of a mass poliomyelitis vaccination program on the occurrence of enteroviruses in seawater. AB - A mass poliomyelitis immunization program undertaken in a coastal city correlated with a significant increase in the levels of vaccinal-type strains of poliovirus recovered in the surrounding seawater. These vaccinal strains were detected together with other wild strains regularly isolated that did not exceed the levels usually detected. The need for a serious monitoring of uncontrolled sewage discharges into the sea is discussed. PMID- 3033953 TI - [Serum antibodies to the hepatitis A virus in persons following a history of the infection]. AB - Enzyme immunoassay was used for titration of serum antibodies in subjects with a history of clinically pronounced or asymptomatic hepatitis A infection. Titers of hepatitis A virus antibodies (anti-HAV) essentially decreased during 3-4 years after the disease, then the rate of this decrease slowed down and antibody titers stabilized at low levels. After clinically pronounced hepatitis A anti-HAV levels were considerably higher than after the asymptomatic form of this infection. PMID- 3033955 TI - [Pathogenesis of persistent and chronic forms of tick-borne encephalitis (experimental study)]. AB - A long-term experiment was conducted to study various aspects of the pathogenesis of persistent and chronic tick-borne encephalitis (TBE). Virological, serological, pathomorphological, electron microscopic and immunofluorescent techniques have been utilized in this study. Persistent TBE infection of Syrian hamsters examined over the period from 40 days to 2 years was characterized by the presence of virus-specific antigens in the organs and of specific antibodies in the blood serum. The persisting TBE virus was found to be predominantly localized in the central nervous system and spleen. Nerve cells underwent ultrastructural changes which were characteristic of flavivirus infection and related to the morphogenesis of viral particles. The authors have developed an experimental model of a primary progressive form of TBE with early and late manifestations of clinical symptoms of the disease. PMID- 3033954 TI - [Intrahospital outbreak of enterovirus encephalitis]. AB - The authors examined 9 children aged from 13 months to 9 years who, while being in a general hospital, contracted enteroviral encephalitis induced by Coxsackie B3 virus with a possible involvement of Coxsackie B1 virus. This report on a group hospital outbreak of enteroviral encephalitis appears to be the first of the kind in the available literature. The authors discuss the possibility of enteroviral encephalitis and Economo's encephalitis being identical in nature. PMID- 3033956 TI - [Diphtheria polyneuropathy in adults]. AB - Neurological manifestations of diphtheria have been studied in 32 adults. The main syndromes of peripheral nervous system damage of diphtheria etiology are presented. The pathogenic mechanisms of the development of neurological complications and the efficacy of multiple-modality restorative therapy were considered. PMID- 3033957 TI - [Diagnosis of variants of neuropathies using the standardized methodology of quantitative analysis of the interference EMG]. AB - In examining 36 patients with neuropathies the authors used standardized leads and a quantitative analysis of the interferential EMG and established the criteria for the differential diagnosis of radiculopathies, neuropathies in particular. Comparison with the findings of analysis of potentials of motor units showed that a "myopathic" shift revealed in some patients was due to stage characteristics of denervation and re-innervation process. PMID- 3033958 TI - [Multiple sclerosis associated with myasthenia]. AB - Two patients with a rare combination of generalized myasthenia (M) and multiple sclerosis (MS) have been studied. The development of both diseases in the same patient, despite the rarity of such reports in the world literature, does not appear accidental and may be due to the common nature of pathogenetic mechanisms. The authors believe that the fundamental homogeneity of the diseases lies in impairment of immunological homeostasis characteristic of both autoimmune diseases. Myasthenic manifestations in MS may widely vary from mild subclinical forms of impairment of neuromuscular conductivity to severe generalized forms of M. The authors discuss possible mechanisms of blockage of the neuromuscular conductivity in cases of association of the above nosological entities. PMID- 3033959 TI - [Isolation and characterization of ouabain-resistant mutants of Chinese hamster cells (Wg3-h)]. PMID- 3033960 TI - [Ultrastructural changes of the human gastric cancer cell line MGc 80-3 caused by HpD-laser in vitro]. PMID- 3033961 TI - [Ultrastructural observation of xenotropic C-type viruses invading rat hepatoma (BERH-2) after passage in nude mice]. PMID- 3033962 TI - [Studies on the functional development of Rohon-Beard cells and their relation to early embryonic behavior]. PMID- 3033963 TI - [Comparison of the DNA and restriction DNA fragments of the rat liver and hepatoma BERH-2]. PMID- 3033964 TI - Biosynthesis of sterols and dolichol in human hepatomas. PMID- 3033965 TI - Icterogenic hepatocellular carcinoma and polygonal cell carcinoma with fibrous stroma (fibrolamellar hepatocarcinoma). AB - Hepatocellular carcinoma (HCC) associated with obstructive jaundice by direct invasion or migration of tumor tissue into the biliary system has been described as icterogenic hepatocellular carcinoma (IHCC) or icteric type hepatoma. Fifty eight such cases have been reported in literature since 1947. Curative treatment and prognosis depend directly on early appearance of icterus and its correct interpretation. Recently, attention has been paid to polygonal cell carcinoma with fibrous stroma (PCFS) as a well differentiated, low grade and transient type of HCC, probably distinguishable from classic HCC by etiological and epidemiological features. The combination of IHCC and PCFS is very rare; to our knowledge only one such case has been reported yet. We had the opportunity to observe and treat such a case, which will be reported in this article and will be compared with cases described in the literature. PMID- 3033967 TI - [Value of nuclear magnetic resonance tomography and first-pass radionuclide ventriculography in cardiomyopathies]. AB - 12 patients with cardiomyopathy were examined by MRT and first pass angiocardiography. MRT provides detailed images of cardiac anatomy and abnormalities without using any contrast medium. The first pass method is an excellent complement to MRT. The functional imaging describes regional wall motion of myocardium exactly. Both non-invasive methods are useful for the diagnosis of cardiomyopathy. PMID- 3033966 TI - Effect of oral morphine and naloxone on pituitary-adrenal response in man induced by human corticotropin-releasing hormone. AB - To further investigate the role of opioids in the regulation of the pituitary adrenal axis we studied the effect of morphine and naloxone on human corticotropin-releasing hormone (hCRH)-induced ACTH, immunoreactive (ir) beta endorphin, and cortisol release in normal subjects. Protocols: 1. 30 mg of a slow release preparation of morphine or placebo was given orally 3 h prior to administration of hCRH (0.1 mg iv) (N = 7). 2. Naloxone (4 mg as bolus iv) or placebo was given 5 min prior to hCRH (N = 7). 3. Naloxone (4 mg iv as bolus followed by a continuous infusion of 6 mg over 75 min) or placebo was started 15 min prior to hCRH (N = 6). hCRH was injected at 11.00 h (protocol 1, 2) or at 17.00 h (protocol 3). Oral morphine not only suppressed basal hormone levels (P less than 0.02), but also the peak response to hCRH compared with placebo (cortisol: 270 +/- 50 vs 559 +/- 80 nmol/l; ACTH: 5.1 +/- 1.5 vs 13.1 +/- 2.7 pmol/l; ir beta-endorphin: 48.5 +/- 8.7 vs 88 +/- 14 pmol/l; mean +/- SEM, P less than 0.02). Similarly, the maximum incremental changes and the area under the curve were significantly reduced for all three hormones compared with placebo (P less than 0.05). After 4 mg of naloxone in the morning, no significant hormonal changes in response to hCRH were observed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3033968 TI - Halothane, enflurane and isoflurane anaesthesia for adenoidectomy in children, using two different premedications. AB - In 48 children subjected to adenoidectomy, comparisons of airway problems, heart rates, cardiac arrhythmias, ventilation and stress hormone reactions were studied during halothane, enflurane and isoflurane anaesthesia. Sixteen children were anaesthetized with either of the three agents and eight patients in each group received diazepam 0.25 mg kg-1 and atropine 0.015 mg kg-1 rectally (DA) as premedication and the remainder diazepam 0.5 mg kg-1, morphine 0.15 mg kg-1 and scopolamine 0.01 mg kg-1 (DMS) rectally. All children were intubated and breathing spontaneously. Equianaesthetic inspired concentrations of halothane, enflurane and isoflurane were used. Airway problems were of the same magnitude during halothane and isoflurane anaesthesia but were less frequent with both agents compared with enflurane anaesthesia. DMS reduced the number of airway reactions in all groups. Respiratory rates were uninfluenced by anaesthesia, intubation and surgery during enflurane anaesthesia. Cardiac arrhythmias were less frequent with enflurane and isoflurane than with halothane. Plasma ACTH and cortisol were similar with all three agents. During induction of anaesthesia in the DA-premedicated halothane group, however, plasma catecholamines were higher than in the group which received DMS, in contrast to the findings during enflurane and isoflurane anaesthesia. The DMS premedication decreased the response of plasma ACTH, cortisol and plasma catecholamines to surgery. PMID- 3033970 TI - Identification of adipose tissue primordia in perirenal tissues of pig fetuses: utility of phosphatase histochemistry. AB - Perirenal adipose tissue samples were obtained from fetuses removed from pregnant (crossbred) sows at 3 stages of gestation (70, 90 and 110 days). Phosphatase histochemistry, succinate dehydrogenase (SDH) histochemistry and factor VIII antigen immunocytochemistry were conducted on fresh-frozen cryostat sections. Age associated changes in nucleosidediphosphatase (NDPase) reactions in the arteriolar system were correlated with the morphological development of the medial layer of arterioles and arteries. For instance, a strong NDPase reaction in small arterioles was associated temporally with the assumption of a normal smooth-muscle cell morphology and arrangement in the medial layer. Age-associated changes in blood vessel reactions for factor VIII antigen and alkaline phosphatase activity were not correlated with morphological development. In the youngest fetuses, alkaline phosphatase activity was evident in large and small arterioles, but in the oldest fetuses, alkaline phosphatase activity was restricted to the smallest arterioles and vessels associated with them. Arteriolar differentiation was demonstrable with either adenosine triphosphatase (ATPase) or inosine diphosphatase (IDPase) reactions. Primordial stromal cells around differentiated arterioles were reactive for ATPase but not for IDPase activities. In older fetuses, there were large areas that contained ATPase reactive stromal cells, no adipocytes, differentiated (ATPase and IDPase) arterioles and few capillaries. Positive reactions for SDH were evident in the ATPase-reactive stromal areas that contained no adipocytes. Differentiated adipocytes were SDH- and ATPase-reactive. These data illustrate the utility of differential phosphatase histochemistry to identify adipose tissue primordia. PMID- 3033969 TI - Hormonal and metabolic responses to cardiac surgery with sufentanil-oxygen anaesthesia. AB - The effects of sufentanil, 10 and 20 micrograms kg-1 on the hormonal and metabolic responses to coronary artery surgery were compared in 20 patients. The most important finding was that the changes in circulating beta-endorphin, ACTH, cortisol, GH, glucose, lactate and glycerol concentrations during and after cardiac surgery were similar with both doses of sufentanil. Although sufentanil prevented a significant increase in plasma beta-endorphin, ACTH and cortisol values until 6 h after cardiopulmonary bypass (CPB), a significant increase in GH secretion occurred with the onset of CPB. Plasma insulin concentrations declined significantly after 30 min CPB, but recovered after 60 min CPB with the restoration of normothermia. Blood glucose values did not change during surgery before CPB, but started to rise with the onset of CPB and continued to increase significantly in the postoperative period. Changes in blood lactate and plasma glycerol concentrations primarily reflected the load of CPB and the effects of heparin, respectively. The results show that increasing the dose of sufentanil up to 20 micrograms kg-1 does not result in better suppression of the endocrine and metabolic changes associated with cardiac surgery. PMID- 3033971 TI - Electron-microscopic cytochemistry of the catecholaminergic innervation of ACTH containing neurons in the rat hypothalamic arcuate nucleus. AB - The synaptic relationship between catecholamine terminals and adrenocorticotropic hormone (ACTH)-containing neurons in the arcuate nucleus (AN) of the rat hypothalamus was investigated by electron microscopy, using ACTH immunocytochemistry combined with autoradiography after 3H-dopamine (3H-DA) injection or 5-hydroxydopamine (5-OHDA) uptake in the same tissue section. ACTH like (ACTH-LI) immunoreactive nerve cell bodies and fibers received synaptic inputs by axon terminals labeled with 3H-DA or 5-OHDA in the AN. This suggests that catecholaminergic neurons, at least DA- and 5-OHDA-containing neurons, may play an important role in the regulation of ACTH secretion or other functions of ACTH neurons via synapses in the AN of the rat hypothalamus. PMID- 3033972 TI - Quantitative electron-microscopic analysis of the in vivo effects of tetrahydrocannabinol on rat preovulatory follicles. AB - The ultrastructural composition of 4 regions of rat preovulatory follicles was examined using stereological techniques following an intraperitoneal injection of tetrahydrocannabinol (THC), once each day, for 3 days prior to sacrifice on proestrus. Control rats were injected with solvent alone. Changes in nuclear, mitochondrial, lipid droplet and dense-body volume density were measured. No differences in volume density were noted in the theca or antral region of the membrana granulosa. Compared to controls, the mitochondrial volume density decreased in the peripheral region and the corona radiata. The volume density of lipid droplets decreased in the peripheral region. These changes, and their relationship to the roles of these regions in follicular steroidogenesis, are discussed. PMID- 3033973 TI - Neurological disorders in Nigerian Africans: a community-based study. AB - In a Nigerian town with a stable population of 20,000, a door-to-door survey was conducted, using a questionnaire involving a complete census and a simple neurological evaluation which had previously showed a 95% sensitivity and an 80% specificity for detecting neurological disease. Positive responders were evaluated and categorised, using agreed criteria for diagnoses. Nearly 100% cooperation was obtained. Life prevalence ratio for at least one episode of headache was 51/1000. Crude point prevalence ratio for migrainous headache was 5.3/100, and peak age-specific ratio was in the first decade. Prevalence ratio for epilepsy was 533/100,000 and peak age-specific prevalence ratio occurred in the 5-14 years age groups. The prevalence ratio for peripheral nerve disorders was 268/100,000, and age-specific prevalence ratio for tropical neuropathy increased with age. Prevalence ratio for stroke was rather low at 58/100,000, but was probably due to the people's attitude to the disabled elderly and high mortality of stroke which showed annual mortality rate of 70/100,000 which increased with age to 1519/100,000 per year in the eighth decade. Crude prevalence ratios (cases per 100,000) for others are 112 for neurological complications (including sciatica) of spondylosis, 15 each for poliomyelitis, motor neurone disease, development speech disorders, 10 each for syncope, hereditary neuropathies. Parkinson's disease, benign essential tremor, primary cerebellar degeneration, cerebral palsy, mental retardation, organic psychosis (probable intracranial tumor) and 5 each for muscular dystrophy, pyomyositis, spina bifida occulta, alcohol dependence and cerebral malaria. The implications of the findings are important for development of community neurological services in the developing countries. PMID- 3033974 TI - Polyneuropathy in Waldenstrom's macroglobulinaemia. Passive transfer from man to mouse. AB - To support the hypothesis of an immunopathogenesis of polyneuropathy in Waldenstrom's macroglobulinaemia (MW), serum IgM fractions of MW patients were applied intraperitoneally to mice for 17 days. Sections of liver, kidney, M. glutaeus maximus, central nervous system (CNS) and both Nn. ischiadici were examined for IgM, IgG, C3 and as control IgD with PAP-immunostaining. IgM deposits were found in every organ except the CNS. In peripheral nerves larger amounts were visualized in perineurium and endoneural space, whereas myelin lamellae and periaxon did not stain. Therefore, perhaps our investigation reveals a greater permeability of the blood-nerve barrier (BNB) compared with the blood brain barrier (BBB). The involvement of the monoclonal IgM of MW, which has been shown to react in vitro with peripheral nerve constituents, appears possible in the pathogenesis of polyneuropathy. PMID- 3033975 TI - Analysis of F-wave in metabolic neuropathies: a comparative study in uremic and diabetic patients. AB - Motor nerve conduction study along the entire length of the ulnar and tibialis posterior nerves was carried out in 30 diabetics compared with 30 uremic patients and 30 control subjects. The conduction in the proximal and the distal nerve segments was evaluated by the determination of the M and F latencies, MNCV (between the stimulus sites), FWCV (between the spinal cord and the stimulus sites), and F-ratio (conduction time ratio of proximal to distal segment). In both groups of patients the lower limbs appear much more involved than the upper, where the ulnar nerve is more commonly affected in uremic than in diabetic patients. In diabetic neuropathy the motor conduction abnormalities are diffuse over the total length of the nerve, but more marked distally in the ulnar nerve. PMID- 3033976 TI - Viral IgM antibodies in serum and cerebrospinal fluid in patients with multiple sclerosis and controls. AB - Serum and cerebrospinal fluid (CSF) from 28 patients with multiple sclerosis (MS), 14 with other neurological diseases (OND) and 31 control subjects with tension headache were analysed for presence of IgM antibodies against measles, mumps and varicellae zoster by a specific enzyme-linked immunosorbent assay (ELISA). This technique excluded false positive reaction due to possible presence of rheumatoid factor. Twelve of the 28 patients with MS had IgM antibodies in serum and 4 in CSF, the latter always being accompanied by presence of corresponding IgM antibodies in serum. Six patients had mumps specific IgM, 5 had measles specific IgM and 3 varicellae specific IgM. In 2 patients, viral IgM antibodies were demonstrated in serum against 2 different viruses. Among the 14 OND patients, one with Wilson's disease had demonstrable serum IgM varicellae antibodies and one with radicultis had elevated serum and CSF measles and varicellae IgM antibodies. Among 31 controls, 2 had IgM antibodies in serum, one against varicellae and one against mumps. No correlations were found between viral IgM antibodies and CSF IgM index, serum IgM levels or blood-brain barrier state. Our data show that MS may be accompanied by a systemic IgM response against the 3 viruses tested, occasionally against 2 of the 3 different viruses simultaneously. The occurrence in MS of virus-specific IgM may be a reflection of viral reactivation and/or polyclonal B cell activation. PMID- 3033977 TI - A case of adult neuronal ceroid-lipofuscinosis with the appearance of membranous cytoplasmic bodies localized in the spinal anterior horn. AB - An autopsy case of adult neuronal ceroid-lipofuscinosis was examined. The clinical picture was characterized by gait disturbance, bulbar palsy and dementia. Histopathologically, diffuse neuronal loss was found throughout the central nervous system. The remaining neurons, predominantly in the motor nuclei of the spinal cord and brain stem, were swollen with storage material. Observed under the electron microscope the storage material showed various ultrastructures, such as lipofuscin-like bodies, pleomorphic lipid bodies, curvilinear profiles and finger-print profiles, in different regions of the central nervous system. In the ballooned neurons of the spinal anterior horn, many membranous cytoplasmic bodies and curvilinear profiles were intermingled within the same cell and were continuous with each other. Biochemically, N-acetyl neuraminic acid content was significantly increased in the spinal anterior horn. These findings suggest the localized increase of ganglioside in that region. PMID- 3033979 TI - Concentration of angiotensin-converting enzyme in tears of patients with sarcoidosis. AB - The concentration of angiotensin-converting enzyme (ACE) was studied in 39 patients with sarcoidosis, 6 of whom had active uveitis, 7 patients with non sarcoid uveitis and 36 healthy controls. ACE concentration in tears was also compared with total protein concentration in tears in order to exclude the effect of varying dilution of tears at sampling. Mean tear ACE concentration and ACE/protein ratio were higher in patients with sarcoidosis than in controls. There were no significant differences in tear ACE concentration or ACE/protein ratio between sarcoidosis patients with uveitis and those with no eye involvement. Tear ACE concentration and ACE/protein ratio did not correlate significantly with serum ACE concentration. It is concluded that the mean concentration of tear ACE and ACE/protein ratio are elevated in sarcoidosis, but that this elevation is independent of any eye involvement. PMID- 3033978 TI - Virus spread and initial pathological changes in the nervous system in genital herpes simplex virus type 2 infection in mice. A correlative immunohistochemical, light and electron microscopic study. AB - Mice were infected by the vaginal route with the MS strain of herpes simplex virus type 2 (HSV-2). Serial vaginal cultures were used to confirm infection and to select mice for this study. Two mice were killed by perfusion on days 2-6 post infection (p.i.) and lumbar and sacral cord with cauda were fixed and embedded for electron microscopy. Semithin Epon-sections were stained for viral antigen using a rabbit anti-HSV-2 antiserum and the Avidin-Biotin (ABC) method. Thin sections from antigen-positive blocks were examined by electron microscopy, and the number and types of infected cells detected by these two methods were compared. A good correlation was found between detection of infected cells by these methods. Infected cells included neurons of dorsal root ganglia and spinal cord, satellite cells of dorsal root ganglia, non-myelinating Schwann cells, astrocytes, oligodendrocytes and arachnoidal cells. Infected cells were first detected in the cauda on day 3 p.i. and in the spinal cord on day 5 p.i. The temporal and spatial distribution of infected cells was consistent with neural spread to and within the CNS. The pathological lesions showed a good correlation with the distribution and number of infected cells and are probably due to a direct virus effect. The similar sensitivity of the Epon-ABC method to electron microscopy in detecting infected cells indicates that this method may have useful applications in both experimental and diagnostic work. PMID- 3033980 TI - Resection for macrodactylism of toes. Report of three cases. AB - Three cases of macrodactylism of toes were operated on with a V-shaped resection: the toe, approximately two thirds of the metatarsals, and part of the adjacent interosseous muscles were resected. The cosmetic and functional results were good. PMID- 3033981 TI - Ultracytochemical study of ouabain-sensitive, potassium-dependent p nitrophenylphosphatase activity in the inner ear of the squirrel monkey. AB - The localization of ouabain-sensitive, K+-dependent p-nitrophenylphosphatase (K+ NPPase) activity of the Na+, K+-ATPase complex was studied ultracytochemically in the squirrel monkey inner ear. In the stria vascularis the reaction products showing K+-NPPase activity were limited to the cytoplasmic side of the plasmalemmal infoldings of the marginal cells. In the spiral prominence, a weak reaction was also found on the cytoplasmic process of the stromal cell, while no or little reaction was detected on the spiral prominence epithelium. In the dark cells of vestibular labyrinth the reaction products were observed on the basolateral interdigitation of the plasmalemma. In contrast, no reaction was observed on the apical cell surface. K+-NPPase activity was most intense in the strial marginal cell, followed by the dark cell of the ampulla and the utricle. The present results revealed that the dark cells in the vestibular labyrinth are involved in endolymph homeostasis. PMID- 3033982 TI - Influence of noradrenalin on pterygopalatine ganglionic transmission of the rabbit in vitro. AB - The influence of noradrenalin on pterygopalatine ganglionic transmission was examined in the rabbit in vitro. Electrical activity recorded from the posterior nasal nerve in response to stimulation of the vidian nerve was reversibly inhibited by the superfusion of noradrenalin. This inhibition was counteracted by phentolamine. This result indicates that, through alpha-adrenergic receptors, noradrenalin inhibits transmission in the pterygopalatine ganglion. PMID- 3033983 TI - Tumours of the minor salivary glands. A clinicopathologic study of 243 cases. AB - 243 patients with tumours of the minor salivary glands were analysed clinicopathologically. The palate was the most common location for the lesions. Mixed tumours (pleomorphic adenomas) were the most common benign tumour type. Muco-epidermoid carcinoma and adenoid cystic carcinoma were the dominant types of malignant tumours. For malignant tumours, the overall 3-year survival rate was 84.0%, 5-year 80.2%, 10-year 66.7% and 15-year 53.6%. The overall recurrency rate was 38.9%. The rate of cervical lymph node metastases and distant metastases rate were both 9.2%. PMID- 3033985 TI - Long term survival case of small (oat) cell carcinoma of the rectum. AB - A long term survival case of small (oat) cell carcinoma of the rectum in a 39 year-old female is presented. She complained of anal pain and occasional anal bleeding. The tumor was located at the anterior wall in the lower rectum. Biopsy specimens revealed a carcinoid tumor. She underwent trans-anal local resection for the first time in December, 1980. Macroscopic findings of the resected specimen showed a small nodule, 0.4 by 0.4 by 0.5 cm, with yellowish cut-surface. Microscopically, the tumor deeply invaded the submucosal layer. The appearances were indistinguishable from pulmonary small (oat) cell carcinoma. Since lymphatic permeations were moderately recognized in the tumor, she underwent radical operation (Miles' operation) with lymphadenectomy. Microscopic findings of the resected rectum revealed an intramural metastatic lesion with marked lymphatic permeations in the submucosal layer 2 cm distant from the primary lesion. Up to date, there is no evidence of local recurrence or liver metastasis. Small (oat) cell carcinoma of the rectum easily metastasizes lymphogenously through the lymph system from an early stage of the development. Wide surgical resection will be needed to give a long term survival even if the tumor is extremely small. PMID- 3033984 TI - Diffuse interstitial pulmonary fibrosis and lung cancer. AB - Forty-two cases of lung cancer complicated with diffuse interstitial pulmonary fibrosis (DIPF) were selected from 13,056 autopsy cases. They were divided into primary (Group I and II) and secondary (Group III) DIPF, and histopathologic and clinicopathologic studies were made. The prevalence of lung cancer with primary DIPF was 17% (8/47) which was significantly (p less than 0.01) higher than that without DIPF (7% or 886/13,009). All eight cases of lung cancer in Group I and II were male, and 7 had cigarette-smoking history. The anatomical sites of lung cancer and DIPF were all peripheral in origin, and in 7 cases located primarily in the lower lobes (left:right = 6:1). Adenocarcinoma was the most common type. The study suggests that male DIPF patients with long clinical history should be followed thoroughly for lung cancer. PMID- 3033986 TI - Demonstration of pituitary tissue with 6 cells immunoreactive to pituitary hormones in a sacrococcygeal teratoma. AB - A sacrococcygeal teratoma, containing mature-appearing anterior pituitary tissue, was first reported with the result of an immunohistochemical analysis for pituitary hormones. All kinds of adenohypophyseal endocrine cells were demonstrated in the anterior pituitary tissue in this teratoma. This study revealed that the anterior pituitary tissue being contained together with nerve tissues in a sacrococcygeal mature teratoma has the capacity to produce at least six anterior pituitary hormones. PMID- 3033988 TI - A pathological study on colorectal cancer. From de novo carcinoma to advanced carcinoma. AB - The development and progression of colorectal cancer were studied by examining the mode of proliferation of cancer and adenoma in 806 cases (857 lesions) of colorectal cancers and 12 cases of familial polyposis coli. Colorectal cancer was classified into two groups: those accompanied by intramucosal growth or with polypoid growth and those without polypoid growth. Early cancer in the group without polypoid growth was considered to originate from de novo carcinoma of about 5 mm, showing vascular invasion when reaching the size of about 10 mm and then exhibiting massive submucosal invasion. Their cut-surface closely simulated cancer of ulcerative type without polypoid growth which occupied the greater portion of advanced cancer of the large bowel. In other words, ulcerative type cancer without polypoid growth is considered to arise from de novo carcinoma and this type occupies 70-90% of all colorectal cancers. Since the lesions originating from de novo carcinoma develops from an extremely small intramucosal carcinoma and then leads to a massive submucosal growth, immediate partial resection is recommended. PMID- 3033987 TI - Multiple myeloma, IgA kappa type, accompanying crystal-storing histiocytosis and amyloidosis. AB - An autopsy case of multiple myeloma, IgA kappa type, accompanying systemic crystal-storing histiocytosis and generalized amyloidosis, is reported. Besides multiple destructive lesions in the skeletal bones, nodular myeloma cell infiltrates were scattered in the liver, spleen, and both kidneys. Not only in these lesions but also in the reticuloendothelial organs, crystal-storing macrophages appeared dispersively or in clusters. Electron microscopically, numerous crystalline inclusions contained in the cytoplasm of macrophages were membrane-bound and of variable configuration, comprising of a homogeneous electron-lucid material. Enzyme cytochemically, almost all of the inclusions showed acid phosphatase activity. On the basis of the results obtained from the immunohistochemical, immunofluorescent and immunoelectron microscopic studies, it was considered that the crystalline inclusions stored in the macrophages were derived from IgA kappa immunoglobulin secreted from the myeloma cells and were formed within secondary lysosomes by crystallization during lysosomal digestion and degradation of the ingested immunoglobulin by macrophages. Generalized amyloidosis developed in different sites from those of the crystal-storing histiocytosis and were proven immunohistochemically to belong to AL amyloidosis probably derived from a certain group of A kappa precursor protein. PMID- 3033989 TI - Human normal and neoplastic adrenocortical cells in tissue culture observed by scanning electron microscopy. AB - Human normal and neoplastic adrenocortical cells were incubated under stimulation with ACTH and observed by scanning electron microscopy. Cultured normal adrenocortical cells gathered into small clusters, each cell of which had a polarity or orientation evidenced by two different aspects. In one aspect, the cell surface rounded up and microvilli protruded vertically. Pits were found among or close to the groups of microvilli. In the other aspect, the cell surface was flattened and well-developed microvilli ran horizontally. These two aspects of the cultured cells were thought to correspond to the cell surface facing the intercellular space and that facing the perisinusoidal space, respectively. In incubated cell clusters of adrenocortical adenomas with Conn's syndrome, most cells lost this polarity or orientation and unstimulated cells existed as unit of the clusters, but all adenoma cells reacted to ACTH in the same manner. Microvilli were distributed unevenly. Filopodia were noticed in some cells. Bleb like structures appeared frequently and some of them were about to be extricated from the cell surface as in normal adrenocortical cells. Adrenocortical adenomas with Cushing's syndrome showed remarkable responses to ACTH. Their cell surface was unclean with the adherence of fragmented cytoplasm and bleb-like structures. Horizontally running elongated microvilli were almost indistinguishable from collagen fibrils. Moreover, collagen fibrils were entangled with microvilli. PMID- 3033990 TI - Thalidomide enhances superoxide anion release from human polymorphonuclear and mononuclear leukocytes. AB - The effect of thalidomide on the function of human polymorphonuclear leukocytes (PMNs) and blood monocytes was tested in vitro. The chemotactic and spontaneous migration was not affected by thalidomide between 0.001 and 0.1 mg/ml. PMN oxygen consumption was not changed after pre-incubation with thalidomide. However, superoxide anion release upon stimulation of the cells with phorbol-myristate acetate or the chemotactic tripeptide N-f-Methionyl-Leucyl-Phenylalanine was enhanced in a dose-dependent manner after pre-incubation with thalidomide. Both PMNs and monocytes were influenced. Leukocytes from two patients with chronic granulomatous disease (defective in oxidative burst response) remained unable to produce superoxide anion after pre-incubation with thalidomide. Thus, we suggest that thalidomide primes the phagocytes to respond with an enhanced superoxide anion release upon stimulation. These findings do not explain the anti inflammatory effect of thalidomide but could have relevance in conditions with quantitative defects in phagocyte oxidative responsiveness. PMID- 3033991 TI - [Effects of dimethyltrilobine iodide on neuromuscular transmission]. PMID- 3033992 TI - [Comparative studies on antiviral activity of several TDA analogs against HSV-2]. PMID- 3033994 TI - A search for a model tissue for studying effects of thiazide diuretics. AB - A search was made for a model tissue of NaCl absorption which would be sensitive to inhibition by diuretics of the thiazide type. A lack of such a model through the years has hampered the analysis of the cellular mechanism of action of this important class of drugs. Using the short-circuit current technique, the urinary bladders of the toads Bufo spinosus and Bufo marinus, and the frog Rana temporaria were investigated regarding the effects of various thiazides. These bladders have NaCl absorptive properties similar to those of the distal renal tubules, and are claimed to be sensitive to the inhibitory effect of thiazides on sodium transport. The short-circuit current (SCC), which is representative of the sodium transport across the epithelium, was reduced by cyclopenthiazide and polythiazide, but only at high concentrations (above 0.1 mM). To rule out the possibility that this was an unspecific effect, attempts were made to block the effect by the 'thiazide blocker' Ex 4877, but without success. This finding, together with the fact that dose-response curves were difficult to obtain, would indicate that these epithelia are not suitable for the stated purpose. Preliminary studies were also conducted on the urinary bladder of the plaice, Pleuronectes platessa, which has a different system of NaCl absorption that is claimed to be rapidly and reversibly inhibited by thiazides. Polythiazide, added to both sides of the bladder, had no effect on SCC. PMID- 3033993 TI - Inhibition by dopamine of the neurotransmission in the rat vas deferens. AB - The effects of alpha-adrenoceptor and dopaminoceptor agonists and antagonists were investigated on presynaptic receptors in the prostatic portion of rat vas deferens. The variable studied was the early component (250 msec) of the motor response elicited by field stimulation (single pulses). All the experiments were carried out in the presence of cocaine 30 mumol/l and hydrocortisone 28 mumol/l so as to block the sites of amines loss and 1-propranolol 0.3 mumol/l to block beta-adrenoceptors. Clonidine, noradrenaline (NA) and dopamine (DA) inhibited the motor response in a concentration-dependent manner. DA was 10 and 10(4) times less potent than NA and clonidine respectively. The selective D2 agonist, LY 141865, failed to inhibit the motor response even at a high concentration (30 mumol/l). Yohimbine (0.1, 0.3 and 1 mumol/l) antagonized competitively the effect of clonidine, NA and DA showing similar - log KB values (7.57; 7.68 and 7.09 respectively). Likewise, idaxozan (0.03 mumol/l) blocked the inhibitory effect of DA in the same order of potency (- log KB = 7.81). On the other hand, pimozide 0.21 mumol/l and Schering 23390 3 mumol/l antagonized the inhibitory effect of DA, showing a lower potency than the other antagonists. Taken together, these findings do not support the hypothesis that DA activates a specific population of prejunctional dopaminoceptors to inhibit the motor response elicited by field stimulation in the presence of cocaine, hydrocortisone and 1-propranolol in the prostatic portion of the rat vas deferens. Instead, the population of prejunctional alpha 2-adrenoceptor may be involved. PMID- 3033995 TI - The disappearance of adenosine from blood and platelet suspension in relation to the platelet cyclic AMP content. AB - Adenosine exerts anti-aggregatory effects on human platelets in vitro, probably by increasing intraplatelet levels of cyclic AMP. In addition, adenosine prevents platelet loss in vivo. We have studied the relationship between the concentration of adenosine in the platelet media and the level of cAMP. In PRP, exogenous adenosine (2-16 microM) was eliminated with a half-life close to 5 min. Approximately half of the added adenosine was deaminated (blocked by 1-2 microM EHNA), and half was eliminated by uptake into platelets (blocked by 2 microM dipyridamole). In whole blood the half-life for adenosine was much shorter, about 15 s. Addition of adenosine deaminase (0.3 microgram ml-1) to PRP resulted in a measured half-life for adenosine approximating that of whole blood. In PRP where adenosine was eliminated as quickly as in whole blood, the adenosine-mediated stimulation of cAMP was 35% lower than in PRP, and the cAMP response lasted 2 min versus 15 min in normal PRP. These results suggest that the magnitude and duration of adenosine's effect on platelets are markedly overestimated by studying platelet suspensions. In blood, the effect of adenosine is smaller in magnitude and very transient. The possibility is discussed that the action of adenosine in vivo on blood platelets can therefore be quite local. PMID- 3033996 TI - The dopamine-gamma aminobutyric acid interaction in the striatum of the rat is differently regulated by dopamine D-1 and D-2 types of receptor: evidence obtained with rotational behavioural experiments. AB - The effects of GABA antagonists on apomorphine- and pergolide-induced rotational behaviour were studied with models combining intracerebral and systemic pharmacological treatments. Whether given systemically or intrastriatally to 6 hydroxydopamine-lesioned rats, the GABA antagonist picrotoxin inhibited the rotational responses produced by s.c. administration of the dopamine (DA) D-1/D-2 agonist apomorphine while it enhanced the rotational behaviour produced by the DA D-2 agonist pergolide. Following unilateral injection of picrotoxin or bicuculline into the striatum of naive rats, apomorphine produced ispsilateral rotation, while pergolide produced contralateral rotation. These contrasting effects are compared to the behavioural responses produced by intracerebral administration of GABAergic drugs alone. Intrapallidal injection of picrotoxin produced contralateral rotational behaviour which was independent of pallido nigral pathways. Contralateral rotation was also produced by GABA agonists, but only following intranigral injections. The results are discussed in terms of differences in the localization of DA D-1 and DA D-2 receptors on striatal GABAergic neurons. The DA D-2 receptor agonist pergolide may induce inhibition of striato-pallidal GABAergic neurons, as well as of a local GABAergic circuit exerting inhibition on a striato--pallidal enkephalinergic pathway. However, the DA D-1/D-2 receptor agonist apomorphine may inhibit striatal interneurons exerting inhibition on a striato-nigral GABAergic projection. Such a neuronal arrangement may explain that striatal DA stimulation increases GABA release from the striato-nigral terminals. PMID- 3033997 TI - [3H] imipramine binding sites are decreased in platelets of chronic pain patients. AB - Tritiated imipramine binding to whole platelets was measured in sixteen chronic pain patients not suffering from major depression and in a control group. Maximum binding was significantly lower in chronic pain patients than in the control group, whereas the binding affinity was not significantly different. PMID- 3033998 TI - Antihypertensive and hormonal effects of lisinopril, a new angiotensin converting enzyme (ACE) inhibitor in patients with renovascular hypertension. AB - The antihypertensive and hormonal effects of a new ACE-inhibitor, lisinopril (MK 521), was studied in 11 patients with renal arterial stenosis (bilateral in 1). Oral doses exceeding 5 mg a day significantly reduced blood pressure (BP), the maximum fall occurring 6 h after taking the drug. At higher doses (20-80 mg/day) sustained antihypertensive effects persisted for 24 h. Lisinopril was equally effective in lowering supine and standing BP. When the drug was given stepwise in increasing doses, (5, 10, 20, 40, and in 5 cases 80 mg/day) the BP was successively normalized in 10 of 11 patients (supine BP less than 90 mmHg). 3 patients with low renin hypertension (LRH) responded less well to monotherapy on long-term treatment with lisinopril than the others. A significant increase in heart rate was observed, initially and after 1 month of treatment. After 5 days treatment with increasing doses the plasma concentrations of angiotensin II (AII) and aldosterone (Aldo) fell significantly to very low concentrations. However, on long term treatment (3 months) suppression of AII and Aldo did not always take place. A concomitant decrease in 24 h urinary aldosterone excretion occurred. No changes in renal function or other biochemical tests except for a slight increase in S-K were observed. There were no adverse side-effects. We conclude that lisinopril is an effective and safe medication for renovascular hypertension. PMID- 3033999 TI - Competitive inhibitor binding assay (CIBA) of ACE inhibitors. AB - We describe a new principle for measuring concentrations of pharmacologically active captopril or other angiotensin-converting enzyme (ACE) inhibitors in blood. Serum is incubated with 125-I-labeled ACE inhibitor (substance 351A, a lisinopril analogue, Merck Sharp & Dohme) in a nonequilibrated system, in which label and ACE inhibitor compete for binding to added serum ACE. Free label is separated by adsorption to coated charcoal. Concentration of captopril or other ACE inhibitor is calculated from a standard curve. Results in healthy volunteers showed rapid absorption of captopril with maximal concentration of active drug within 1 h, and fast disappearance within 2.5 h. Stability of captopril was improved by immediate 1:100 dilution of blood samples with assay buffer. In spite of this precaution, analysis should be performed within two days to avoid loss of active drug due to polymerization and protein binding. Samples of other tested ACE inhibitors can be frozen and later analyzed at convenience. The new principle is simple, sensitive, and specific. PMID- 3034000 TI - Antiviral therapy. PMID- 3034001 TI - The syndromic nature of amyotrophic lateral sclerosis. PMID- 3034002 TI - The Mount Vernon stroke service: a feasibility study to determine whether it is possible to apply the principles of stroke unit management to patients and their families on general medical wards. AB - A feasibility study has shown that it is possible to apply the principles of stroke unit management to patients and their families on general wards. Ninety one consecutive stroke patients were managed by a multidisciplinary stroke therapy team which was co-ordinated by a medical registrar, met weekly, and developed a family support service. This was achieved within the pre-existing hospital timetable and staffing level, without providing a specialist 'stroke ward'. (Only 3-4 medical hours a week were needed.) The advantages of the service were: speedy referral to therapists, development of a team approach, provision of better information and psychological support for patients and families, provision of more speech therapy (49%) and occupational therapy (80%) than on other medical wards in other studies. Further research is required to evaluate the effect of the service on specific outcomes of stroke. PMID- 3034003 TI - Ascorbate and cysteine-mediated selective neutralisation of extracellular oxidants during N-formyl peptide activation of human phagocytes. AB - The effects of sodium ascorbate and cysteine (2.5 X 10(-5) M-2.5 X 10(-4) M) on the intensity and profile of luminol-enhanced chemiluminescence, superoxide generation, extracellular myeloperoxidase (MPO) activity and auto-iodination were measured in purified human polymorphonuclear leukocytes activated by the leukoattractant FMLP in vitro. Chemiluminescence studies were also performed using a whole-blood method. Cysteine (10(-4) M-2.5 X 10(-4) M) and ascorbate (2.5 X 10(-5) M-2.5 X 10(-4) M) caused significant inhibition of the early extracellular peak of FMLP-activated chemiluminescence and increased the intensity of the later occurring intracellular peak in both PMNL and blood. At the same concentrations both agents scavenged superoxide released by FMLP activated PMNL, inhibited oxidant generation by extracellular MPO and decreased FMLP-induced auto-oxidation of PMNL. Administration of a single 1 gram oral dose of ascorbate to adult human volunteers was associated with significant reduction and enhancement respectively of the extracellular and intracellular luminol enhanced chemiluminescence responses of FMLP-activated blood. These results show that the water soluble anti-oxidants cysteine and especially ascorbate selectively neutralise the reactivity of harmful reactive oxidants released by phagocytes, whilst the intracellular generation of antimicrobial oxidants remains intact. PMID- 3034005 TI - Haemonchus contortus: lipid biosynthesis from 14C-labelled palmitic acid and sodium bicarbonate. PMID- 3034004 TI - In vitro fibrinolytic activity and viability of rat alveolar macrophages treated with inflammation generating mineral dusts. AB - Rat alvolar macrophages demonstrated plasminogen dependent fibrinolysis in vitro which was inhibited to varying degrees by the addition of zymosan, the non-toxic particulate titanium dioxide, and the toxic dusts quartz and chrysotile asbestos. Assessment of viability suggested that the inhibition produced by zymosan and titanium dioxide could be accounted for by cytotoxic effects but in the case of quartz and chrysotile asbestos there was evidence that stimulation of fibrinolysis preceded cell death. Zymosan, which caused no observeable enhancement of alveolar macrophage fibrinolysis was found to markedly stimulate peritoneal macrophage fibrinolysis. The choice of assays of cell function to assess the action of toxic dusts are discussed. PMID- 3034006 TI - Incorporation of carbon from 14C-labelled precursors into major chemical fractions of Haemonchus contortus in vitro. PMID- 3034007 TI - Abdominal case of the day. PMID- 3034008 TI - Abdominal CT in the staging of small-cell carcinoma of the lung: incidence of metastases and effect on prognosis. AB - CT studies of the abdomen performed on 72 patients with small-cell carcinoma of the lung were retrospectively reviewed to assess the role of abdominal CT in staging. Forty-four of the 72 patients had extensive disease, defined as disease extending beyond the confines of one hemithorax, plus or minus mediastinal or ipsilateral supraclavicular disease or ipsilateral pleural effusion. Initial staging abdominal CT revealed one or more sites of metastatic disease in 26 (59%) of these 44 patients, while 18 patients had normal initial CT examinations. Statistical analysis of patients with extensive disease revealed a significant increase in complete therapeutic response (p = .0054) and in the length of survival (p = .001) among those who had extensive disease without abdominal metastases as compared with those who had abdominal metastases on their initial abdominal CT examination. The development of new or recurrent abdominal metastases in general or in specific organs on follow-up scans obtained in 35 patients did not significantly decrease their survival time as compared with that of patients without such metastases. Our findings suggest that CT of the abdomen is beneficial in the initial staging of patients with small-cell carcinoma of the lung and provides prognostic information concerning response to therapy and length of survival. PMID- 3034010 TI - Dyke award. Harmonic modulation of proton MR precessional phase by pulsatile motion: origin of spinal CSF flow phenomena. AB - The effects of pulsatile motion on MR imaging of spinal CSF were quantitatively evaluated with a spine phantom that simulated spinal CSF pulsation. Two fundamental interdependent pulsation flow phenomena were observed: variable reductions in signal intensity of pulsatile CSF (signal loss) and spatial mismapping of this signal beyond the confines of the subarachnoid space (phase shift images). Phase-shift images were observed as multiple regions of signal intensity conforming morphologically to the subarachnoid space but displaced symmetrically from it along the phase-encoding axis, either added to or subtracted from stationary signal intensity. Both CSF pulsation flow phenomena occurred secondary to harmonic modulation of proton precessional phase (temporal phase shift) by the unique pulsatile motion of spinal CSF when the repetition time was not an integral multiple of the pulsation period. Each flow phenomenon was analyzed with the spine phantom independently to control individual imaging and physiologic parameters including imaging plane, repetition time, echo time, slice thickness, number of echoes, number of excitations, CSF pulsation amplitude, and CSF pulsation period. In the axial plane, signal loss was present on both first- and second-echo images and was more pronounced with larger pulsation amplitudes and smaller slice thicknesses. A quantitative relationship between these two parameters allowed the prediction of CSF pulsation amplitude when the slice thickness was known and the CSF signal intensity was measured. In the sagittal plane, signal loss was present on first-echo images, was more pronounced with larger pulsation amplitudes, and underwent incomplete even-echo rephasing on second-echo images. Phase-shift images were influenced by the relationship between repetition time and CSF pulsation period. They were partly eliminated on sagittal but not on axial second-echo images because of incomplete even-echo rephasing. Both signal loss and phase-shift images were completely eliminated with CSF gating or pseudogating, indicating the rationale for gating during clinical spinal MR. The clinical significance of these findings is that awareness of the existence of spinal CSF pulsation flow phenomena avoids diagnostic confusion, whereas understanding their etiology provides a rational approach, such as CSF gating, to eliminate them. PMID- 3034009 TI - Neonatal hydronephrosis in the era of sonography. AB - During a 6-year period (1979-1985), 142 neonates with significant hydronephrosis were seen. Seventy-eight percent of the cases were discovered on fetal screening during obstetric sonography. Maternal/fetal intervention was virtually never indicated and most babies were asymptomatic. The most common conditions found were obstruction of the ureteropelvic junction (41%), obstruction of the distal ureter (usually primary megaureter) (23%), upper-pole hydronephrosis associated with duplex anomalies (13%), and posterior urethral valves (10%). Seventeen neonates with multicystic dysplastic kidney were seen (three per year or one for every eight with hydronephrosis). In comparison, during the 30-year period, 1947 1977, 146 neonates with significant hydronephrosis were seen. Most cases were discovered because the patients had signs and/or symptoms--either an abdominal mass (an enlarged kidney or bladder) or urosepsis. The three most common conditions were obstruction of the ureteropelvic junction (22%), posterior urethral valves (19%), and ectopic ureterocele (14%). During this period, 53 neonates with multicystic dysplastic kidney were discovered (two per year or one for every three with hydronephrosis). The dramatic increase in the number of neonates found to have hydronephrosis is primarily due to the widespread use of obstetric sonography and concomitant fetal screening. The pattern of causes before 1979 represented the incidence of symptomatic lesions. The current pattern more accurately reflects the true incidence of congenital anomalies of the urinary tract. PMID- 3034011 TI - Chest-wall invasion by carcinoma of the lung: detection by MR imaging. AB - Nineteen patients with bronchogenic carcinoma were studied by MR imaging to determine the presence of chest-wall invasion. All studies were carried out at 1.5 T, and the results were correlated with chest radiographs or CT scans. All MR studies were interpreted before surgery (13 cases) and without knowledge of the results of other radiologic studies. MR findings indicative of chest-wall invasion included a high-signal focus within the chest wall and/or chest-wall thickening with increased signal on spin-echo (SE) images having a repetition time of 2500 msec and an echo time of 50-100 msec (SE 2500/50-100). In one case, noncontour-deforming high-signal intensity within chest-wall musculature (but no other abnormality) was demonstrated on SE 2500/50-100 images. Coronal or sagittal imaging facilitated identification of tumor contiguity with extrathoracic structures in apical lesions. Contrast differences between normal and invaded chest wall on T2-weighted images were the most helpful in assessing chest-wall involvement. These preliminary observations indicate that MR imaging is useful in the evaluation of chest-wall invasion by carcinoma of the lung. PMID- 3034012 TI - Accumulation of technetium-99m sulfur colloid by hepatocellular adenoma: scintigraphic-pathologic correlation. AB - It is currently believed that hepatocellular adenoma is photon deficient on technetium-99m sulfur colloid scintigraphy because these tumors lack Kupffer cells. In a retrospective review of 13 pathologically proven cases of hepatocellular adenoma with technetium-99m sulfur colloid scintigrams, Kupffer cells were present in all 13 cases. We observed uptake of the radiocolloid by the hepatocellular adenoma in three cases (23%), and there were no histologic differences between the tumors with uptake and the ones without it. We conclude that the currently accepted reason for the lack of technetium-99m sulfur colloid uptake within hepatocellular adenoma is incorrect, and an explanation other than a lack of Kupffer cells is responsible for the photon-deficient appearance in the majority of cases of hepatocellular adenoma. Further, because hepatocellular adenoma may have technetium-99m sulfur colloid uptake in a significant percentage of cases, it should be added to focal nodular hyperplasia in the differential diagnosis of a hepatic mass with uptake by technetium-99m sulfur colloid. PMID- 3034013 TI - Soft-tissue masses: histologic basis for decreased signal (short T2) on T2 weighted MR images. AB - Most soft-tissue masses and tumors of various etiologies and histologies have high signal intensity on T2-weighted pulse sequences (long T2). Of 47 soft-tissue masses, seven had a low signal (short T2) on T2-weighted pulse sequences. All seven masses were tumors, and histologic review showed that their composition differed from that of the other 40 lesions with a long T2 in that the seven masses were relatively acellular and had more collagen. The tumors with a short T2 included one malignant and six benign soft-tissue tumors. Malignant fibrous histiocytoma and aggressive fibromatosis showed paradoxical signal intensities in that they showed both long and short T2. All of the tumors with low signal intensity on T2-weighted images had significant fibrous elements and marked hypocellularity. This study suggests that the less commonly encountered short T2 may be seen in both benign and malignant soft-tissue lesions. A part of the explanation for the low signal on T2-weighted sequences appears to be the relative acellularity and abundant collagen of these tumors in comparison with those that have the same histologic diagnoses but show a high signal. The histologic composition of the tumor rather than the histologic diagnosis appears to influence the MR signal on T2-weighted sequences. PMID- 3034015 TI - A symposium: Ramipril--a new angiotensin converting enzyme inhibitor. May 2-4, 1986, Brussels, Belgium. Proceedings. PMID- 3034014 TI - Nonpalpable breast cancer. AB - Although mammography is the most reliable method for the early detection of breast cancer in asymptomatic women and in women with equivocal signs or symptoms, this screening tool is still underutilized in the United States. Precise biopsy of suspicious nonpalpable findings is facilitated by mammographic localization. PMID- 3034016 TI - Efficacy, tolerance and hormonal effects of a new oral angiotensin converting enzyme inhibitor, ramipril (HOE 498), in mild to moderate primary hypertension. AB - The immediate (0 to 24 hours) and long-term (4 weeks) hypotensive effects of a new long-acting angiotensin converting enzyme inhibitor, ramipril (HOE 498), as well as adverse effects and tolerance, were evaluated in 34 patients with primary hypertension. Further, effects on serum and urinary aldosterone and circulating angiotensin II concentrations were measured. After short- and long-term administration of 5 or 10 mg of ramipril, the mean blood pressure was significantly lowered compared with placebo. The mean maximum decrease in blood pressure was noted 4 to 8 hours after administration of ramipril once daily. Sustained blood pressure reduction was achieved after 4 weeks of treatment. Serum concentrations of aldosterone and plasma levels of circulating angiotensin II were reduced for up to 12 hours after drug intake, and tended to return to pretreatment levels at 24 hours. Serum angiotensin converting enzyme activity was markedly suppressed for more than 24 hours after a single dose of 5 or 10 mg ramipril. No subjective or objective adverse effects were noted, and the tolerance to the drug was very good. PMID- 3034017 TI - A double-blind study to compare the efficacy, tolerance and safety of two doses of the angiotensin converting enzyme inhibitor ramipril with placebo. AB - In a randomized, double-blind trial, 2 doses of ramipril (2.5 and 5 mg once daily) were compared with placebo in patients with mild to moderate essential hypertension. A 2-week placebo run-in phase was followed by 4 weeks of treatment. Eighty-six patients entered the study and 17 withdrew during the course of the study. Both doses of ramipril appeared to be more effective than placebo in reducing blood pressure, but significant differences between 2.5 mg of ramipril and placebo were not found in any statistical analyses. In the endpoint analyses (taking the last measurement from each patient), the patients receiving 5 mg of ramipril had significantly larger decreases in blood pressure than the patients receiving placebo (t tests: standing systolic, p less than 0.001; supine diastolic, p less than 0.05; standing diastolic, p less than 0.05) and also than the patients receiving 2.5 mg of ramipril (standing systolic, p less than 0.05). It appears from the results of this study that the minimum effective dosage of ramipril is 5 mg once daily. No clinically relevant side effects or clinically relevant changes in laboratory values were observed. PMID- 3034018 TI - Comparative double-blind study of ramipril and captopril in mild to moderate essential hypertension. AB - The angiotensin converting enzyme inhibitors ramipril and captopril were administered in doses of 10 mg once daily and 50 mg twice daily, respectively, to patients with mild to moderate essential hypertension. After a 4-week single blind placebo washout period, patients were treated for 12 weeks with 1 of the drugs under double-blind conditions. Patients who did not respond after 6 weeks of treatment were given 50 mg hydrochlorothiazide concomitantly. The ramipril group showed greater decreases in blood pressure compared with baseline values: 20.1/14.9 mm Hg (ramipril) compared with 16.5/13.5 mm Hg (captopril). A further 6 weeks of treatment lowered the blood pressure even more: 22.5/20.0 mm Hg (ramipril) compared with 20.5/18.6 mm Hg (captopril). Concomitant hydrochlorothiazide given to nonresponders reduced the blood pressure levels in 24 of 40 patients in the ramipril group and in 20 of 36 patients in the captopril group. At the end of the study the overall response to treatment with ramipril alone and ramipril plus hydrochlorothiazide was 77.1%. The overall response rate in the captopril group was 82.7%. No clinically relevant adverse reaction occurred in any patient. Ramipril given once daily was as effective as captopril given twice daily in lowering blood pressure. Both drugs proved to be safe during treatment for 12 weeks. PMID- 3034019 TI - Systemic and regional hemodynamic profile of five angiotensin I converting enzyme inhibitors in the spontaneously hypertensive rat. AB - The effects of 5 angiotensin I converting enzyme (ACE) inhibitors--captopril, enalapril, perindopril, trandolapril and ramipril--on general and regional hemodynamics were investigated (using radioactive microspheres) and compared in anesthetized adult spontaneously hypertensive rats. The 5 treatments were administered daily by gavage for 8 days in doses inducing identical decreases in arterial blood pressure. This effect was entirely due to a decrease in total peripheral resistance inasmuch as cardiac index was not affected by the 5 ACE inhibitors. In addition, despite their different chemical structures, all exhibited the same regional vasodilator pattern, which thus appears to be related only to ACE inhibition. The vascular beds resistances were decreased in the following order: renal greater than splenic = liver greater than skin greater than total peripheral greater than muscle = brain. Simultaneously, and despite the decrease in perfusion pressure, most regional blood flows and especially renal blood flow were increased. Finally, renal vasodilator effects of ACE inhibitors were observed even after doses that lacked any effect on total peripheral resistance. PMID- 3034020 TI - Efficacy and safety of ramipril (HOE 498) in the treatment of hypertension: dose finding study. AB - An open-label, prospective multicenter trial of ramipril was performed. The agent was administered once daily at an initial dosage of 1.25 mg and this was increased, when necessary, up to 10 mg with intervals of 2 weeks for 8 weeks. Effectiveness in 46 patients with mild to moderate essential hypertension was 28.1% at a dosage of 1.25 mg, 52.2% at 2.5 mg, 69.6% at 5 mg and 78.3% at 10 mg of ramipril alone. In 27 patients receiving baseline therapy with a thiazide diuretic, a subsequent administration of ramipril showed effectiveness in 11.1% for 1.25 mg, 48.1% for 2.5 mg and 70.4% for 5 mg. Adverse effects occurred in 11.7% of patients overall and were not serious. One patient was withdrawn because of severe headache; the other patients tolerated the treatment well. A dosage of 2.5 to 10 mg of ramipril will probably be appropriate for further evaluation of the effectiveness of this agent in a double-blind study. PMID- 3034021 TI - Dose-response relation of the angiotensin converting enzyme inhibitor ramipril in mild to moderate essential hypertension. AB - The effects of various dosage levels of ramipril on blood pressure were examined in a double-blind multicenter clinical trial. Patients with mild to moderate essential hypertension were first entered into a single-blind washout placebo phase of 4 weeks duration. The patients were then randomized to 1 of 3 treatment groups and received either 1.25, 2.5 or 5 mg of ramipril orally once a day for 6 weeks. After 6 weeks of treatment, the diastolic blood pressure had been decreased by 16.0 mm Hg (53 patients), 16.5 mm Hg (54 patients) and 19.9 mm Hg (53 patients) for the 1.25, 2.5 and 5 mg groups, respectively. Systolic blood pressure had decreased by an average of 22.5 mm Hg independent of the dose administered. Diastolic blood pressure was decreased to less than or equal to 90 mm Hg in 64%, 63% and 77% of the patients in the 3 groups, respectively. The differences in these percentages among the groups were not statistically significant. The tolerability of the drug was good; discontinuation of treatment was necessary for 1 patient in the 1.25 mg group and medication was lowered for 2 patients because of adverse reactions. There were no clinically significant changes of any laboratory variables, except for an increase in uric acid serum levels. PMID- 3034022 TI - Ramipril for hypertension secondary to renal artery stenosis. Changes in blood pressure, the renin-angiotensin system and total and divided renal function. AB - The converting enzyme inhibitor, ramipril, 20 mg once daily, was given to 3 hypertensive patients with unilateral renovascular disease. At 1 month, 24 hours after the last dose of ramipril, blood pressure, plasma angiotensin II and converting enzyme activity remained low, and active renin and angiotensin I high. There was no tendency for converting enzyme inhibition to be overcome during 1 month of ramipril therapy. Ramipril caused slight increases in serum potassium and urea, no change in serum creatinine and no consistent changes in the renal vein renin ratio. Ramipril caused little change in renal plasma flow on the stenotic side, but filtration fraction was reduced in 2 patients. There was no serious deterioration in total or individual glomerular filtration rate during ramipril therapy. The drug was well tolerated and there were no serious side effects. Ramipril, given once daily, is likely to be effective in controlling hypertension with renal artery stenosis. PMID- 3034023 TI - Combined treatment of severe essential hypertension with the new angiotensin converting enzyme inhibitor ramipril. AB - Ramipril is a newly synthesized angiotensin converting enzyme inhibitor without a sulfhydryl group in the molecule but with a prolonged duration of action. Efficacy, tolerance and safety of this drug were evaluated in 10 patients with severe essential hypertension. After a treatment period of at least 4 weeks with the conventional antihypertensive drug combination of a diuretic and a beta blocking agent with the vasodilator dihydralazine, their systolic and diastolic blood pressures averaged 161 +/- 6 and 111 +/- 2 mm Hg, respectively. Because diastolic blood pressure during this drug regimen was still greater than 105 mm Hg in all patients, the patients received ramipril initially at single daily doses of 5 mg in addition to their previous medication. The first dose of 5 mg ramipril resulted in a moderate but significant decrease in systolic and diastolic blood pressure in 9 of the 10 patients to 142 +/- 5 and 104 +/- 4 mm Hg (p less than 0.01), respectively, between 3 and 6 hours after drug administration. In 1 patient blood pressure was unresponsive to ramipril and 1 patient complained of nausea and vomiting within the first week of treatment with ramipril. Within the following 8-week treatment period with a once-daily intake of 5 or, if necessary, 10 mg of ramipril, diastolic blood pressure normalized in the remaining 8 patients to less than 90 mm Hg. Systolic and diastolic blood pressure averaged 130 +/- 5 and 83 +/- 2 mm Hg, respectively, at the end of the 8 week treatment period with ramipril. Severe hypotension and reflex tachycardia were not observed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3034024 TI - Acute effects of the new angiotensin converting enzyme inhibitor ramipril on hemodynamics and carotid sinus baroreflex activity in congestive heart failure. AB - The hemodynamic effects of a single dose of 5 mg of ramipril, a new angiotensin converting enzyme inhibitor, were investigated in 10 patients with chronic congestive heart failure. Arterial blood pressure and total peripheral resistance were decreased by approximately 12% without causing reflex tachycardia. A highly significant decrease occurred in mean pulmonary artery and pulmonary capillary wedge pressures. These hemodynamic changes were equally pronounced at rest and during exercise on a bicycle ergometer; the effect was of the same magnitude 5 and 24 hours after medication. Angiotensin converting enzyme activity in plasma was nearly completely inhibited after 5 hours and remained at about 12% of control after 24 hours. Cardiac index, which was normal before treatment, remained unaffected. Thus, ramipril induced a balanced reduction of left ventricular pre- and afterload. The activity of the carotid sinus baroreflex was investigated in 8 of the patients using the neck suction technique before and 24 hours after ramipril. The reflex bradycardia during stimulation of the baroreceptors was significantly increased by ramipril, whereas the decrease in blood pressure remained essentially unaffected. Ramipril induced a selective sensitization of the parasympathetic baroreceptor heart rate reflex without influencing the sympathetically mediated peripheral vasodilatation. This effect may be responsible for the lack of reflex tachycardia in spite of the decrease in blood pressure. PMID- 3034025 TI - Acute hemodynamic, hormonal and electrolyte effects of ramipril in severe congestive heart failure. AB - The response to ramipril, 10 and 20 mg on consecutive days, in 9 patients with severe (New York Heart Association functional class III or IV) chronic congestive heart failure was measured. Hemodynamic cannulae were placed more than 2 days before ramipril administration to ensure a stable baseline. Dietary sodium (40 mmol daily) and potassium (80 mmol daily) were constant before and during the study, and maintenance doses of digoxin and furosemide (80 to 1,000 mg daily) were continued unchanged. Ramipril induced pronounced, sustained decreases in angiotensin converting enzyme activity, angiotensin II and aldosterone levels, and a reciprocal increase in plasma renin activity. Plasma catecholamines, antidiuretic hormone and cortisol levels were not altered. Urinary sodium and potassium excretion diminished, plasma sodium decreased and plasma potassium increased. Plasma urea and creatinine levels increased. Ramipril treatment resulted in a decrease in systemic arterial pressure that was sustained for 24 hours, a decrease in heart rate and an increase in cardiac index, but little change in pulmonary artery pressure or right atrial pressure. Three patients were drowsy after ramipril administration, and 1 patient had a marked, temporary reduction in urine output. It was concluded that ramipril is a potent, long acting angiotensin converting-enzyme inhibitor that is likely to be beneficial in patients with severe cardiac failure. PMID- 3034026 TI - Acute hemodynamic and hormonal effects of ramipril in chronic congestive heart failure and comparison with captopril. AB - Acute hemodynamic and hormonal responses to ramipril in comparison with captopril were studied in 10 patients with moderate to severe congestive heart failure in an open, randomized study. Both drugs were given to 5 patients each in 2 increasing doses on 2 successive days. After 5 mg of ramipril angiotensin converting enzyme (ACE) activity was significantly decreased during 24 hours with a maximum decrease 4 hours after administration. Mean arterial blood pressure decreased from 84 +/- 5 to 62 +/- 5 mm Hg at 4 hours and 71 +/- 4 mm Hg at 12 hours, respectively, after this dose. Capillary wedge pressure decreased from 19 +/- 1 mm Hg to 13 +/- 1 mm Hg at 4 hours with a maximum increase in cardiac output from 3.8 +/- 0.3 liters/min to 4.4 +/- 0.3 liters/min at 2 hours. No significant cardiac effects were present 8 hours after administration. After 10 mg of ramipril, cardiac and hormonal effects showed a quicker onset of action and longer duration compared with the 5 mg dose. Mean arterial pressure decreased to 61 +/- 6 mm Hg. Similar effects were seen after captopril, but with a significantly shorter duration. Mean arterial pressure decreased from 82 +/- 4 mm Hg to 64 +/- 5 mm Hg after 12.5 mg and to 58 +/- 6 mm Hg after 25 mg of captopril. In patients with congestive heart failure ramipril has the hemodynamic profile of a long-acting and potent ACE inhibitor. Significant cardiac effects are present during 4 to 8 hours and ACE activity is still significantly inhibited 24 hours after a single dose of ramipril.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3034027 TI - Effect of converting enzyme inhibitors on tissue converting enzyme and angiotensin II: therapeutic implications. AB - Local tissue renin-angiotensin systems have recently been discovered in various organs, and evidence is accumulating that inhibition of these local renin angiotensin systems may contribute to the actions of converting enzyme (CE) inhibitors. Measurements of CE activity and angiotensin II concentrations revealed that after oral administration of CE inhibitors, CE was inhibited not only in lung vascular endothelium and blood, but also in the heart, kidney, vascular wall, brain and other organs. The functional significance of tissue CE inhibition is suggested first by the antihypertensive effect of brain CE inhibition in spontaneously hypertensive rats, second by the concomitant persistence of blood pressure decrease and CE inhibition in vascular wall and kidney after long-term oral CE inhibitor treatment and third by ex vivo experiments demonstrating marked effects of oral CE inhibitor pretreatment on cardiac function in isolated rat hearts. Local inhibition of tissue renin angiotensin systems may be an important factor involved in the beneficial effects of CE inhibitors in such cardiovascular diseases as arterial hypertension, congestive heart failure and cardiac arrhythmias. PMID- 3034028 TI - Clinical pharmacology of ramipril. AB - Ramipril is a long-acting non-sulphydryl converting enzyme inhibitor that requires cleavage of its ester group to form the active diacid metabolite, ramiprilat. Renal excretion largely determines the drug's duration of action and the dosage should be reduced in patients with renal impairment. Oral ramipril given daily at dosages of 5 mg or more can control blood pressure over a 24-hour period; lower doses may be effective in patients with heart failure inadequately controlled by diuretics alone. No serious idiosyncratic adverse reactions have been reported. Ramipril is one of the most potent long-acting converting enzyme inhibitors developed; it is effective given once daily in the treatment of all grades of hypertension and of heart failure. PMID- 3034029 TI - Pharmacokinetics and effects on the renin-angiotensin system of ramipril in elderly patients. AB - Converting enzyme inhibitors are likely to be prescribed with increasing frequency in elderly patients. The pharmacokinetics of ramipril, a new potent long-acting non-sulphydryl converting enzyme inhibitor, and its effects on blood pressure, plasma renin activity and angiotensin II concentrations were studied in a group of 8 elderly volunteers (mean age 77, range 61 to 84). Circulating concentrations of the active diacid formed from its parent drug were consistently higher in this group despite apparently normal renal function, assessed by serum creatinine and urea concentrations, compared with younger volunteers (age range 21 to 30). The initial dose of ramipril should be lower in older subjects. The study emphasizes the importance of careful extrapolation of data obtained from young volunteers to older subjects. PMID- 3034030 TI - Ramipril: review of pharmacology. AB - Ramipril is a potent orally active converting enzyme inhibitor. Its active metabolite ramiprilat is classified as a reversible, slow- and tight-binding inhibitor. Ramipril lowers blood pressure in various models of hypertension and improves states of acute cardiac failure mainly by suppression of angiotensin II formation. Actions on both vasoconstrictor and volume factors are involved because ramipril causes vasodilation and mild natriuresis but preserves potassium. Sustained inhibition of converting enzyme in target tissues such as vascular wall, kidney and heart may explain cardiovascular changes over the long term. Ancillary effects may possibly emerge from modulation of the sympathetic nervous system but the contribution of bradykinin potentiation remains unclear. PMID- 3034032 TI - Pharmacokinetics, converting enzyme inhibition and peripheral arterial hemodynamics of ramipril in healthy volunteers. AB - The effects of a 10 mg dose of ramipril, a new angiotensin I converting enzyme (ACE) inhibitor, on systemic blood pressure, heart rate, brachial artery blood flow, brachial artery diameter, carotid artery blood flow, carotid artery diameter, forearm vascular resistance, plasma ACE and renin activities and plasma aldosterone were investigated. Ramipril's effects in 6 healthy volunteers on a normal sodium diet were compared with those of placebo over a 24-hour period after oral drug intake in an open cross-over trial. Ramipril inhibited plasma ACE activity, an effect that peaked at 3 to 4 hours and persisted up to at least 24 hours. Plasma renin activity increased from 4 to 12 hours after drug intake and plasma aldosterone was slightly decreased. Systemic blood pressure in the supine position was slightly decreased between 6 and 8 hours after drug intake but heart rate remained unaffected. Ramipril significantly increased brachial artery blood flow, brachial artery diameter and carotid artery blood flow and decreased forearm vascular resistance between 3 and 8 hours after drug administration. These peripheral arterial vasodilating effects were more marked in the muscular resistance vessels and affected both large arteries and arterioles in the brachial vascular territory. A correlation was found between the log of plasma concentrations of ramipril diacid metabolite and the drug-induced plasma ACE activity inhibition and increase in brachial artery blood flow. There was also a correlation between these 2 latter effects. A plasma ACE activity inhibition of 80% was required to induce significant increases in brachial artery blood flow and carotid artery blood flow. PMID- 3034031 TI - Pharmacokinetics of ramipril in the elderly. AB - Ramipril (HOE 498) is a pro-drug of which the main metabolite (HOE 498 diacid or ramiprilat) is a potent angiotensin converting enzyme inhibitor. Thirteen healthy white volunteers (5 females and 8 males, ages 65 to 76 years) participated in a study to investigate the pharmacokinetics of HOE 498 in the elderly. After administration of 10 mg of HOE 498, sequential urine and serum specimens were obtained for assay of HOE 498 and metabolites (HOE 498-glucuronide, diacid, diacid-glucuronide, diketopiperazine and diketopiperazine acid). Side effects, clinical chemistry and hematology were monitored. HOE 498 reached peak concentrations of 62.4 +/- 23.3 ng/ml in serum after 0.7 +/- 0.3 hours. Serum levels decreased with an apparent half-life of 0.9 +/- 0.4 hours. The diacid was rapidly formed in serum, reaching peak concentrations of 40.6 +/- 14.0 ng/ml after 2.0 +/- 0.6 hours and declining with a half-life of 2.2 +/- 0.5 hours. A prolonged terminal phase of serum concentration versus time curve was observed at concentrations less than 1 ng/ml. The mean recovery of HOE 498 and metabolites in urine, up to 26 hours after administration, was 35 +/- 14% of the dose. The apparent half-lives, calculated from urine parameters, for HOE 498 and the diacid were 2.6 +/- 0.9 and 4.0 +/- 1.1 hours, respectively. The mean peak concentration and half-life of HOE 498 in serum are slightly higher in the elderly than in younger volunteers. Complete urinary collection was not possible, but urinary recovery did not seem different from younger volunteers. PMID- 3034033 TI - Baroreflex setting and sensitivity in normal subjects: effects of pharmacologic inhibition of the angiotensin I converting enzyme. AB - Arterial blood pressure, heart rate and the response of these hemodynamic parameters to exogenous norepinephrine were investigated in healthy volunteers (daily sodium intake of 150 mmol) during a control period and after a single oral dose of 5 mg of the angiotensin I converting enzyme (ACE) inhibitor ramipril (HOE 498). Norepinephrine was infused at doses of 0.1, 0.2 and 0.3 micrograms kg-1 min 1, each for 10 minutes, during control and 3 hours after ramipril administration. Exogenous norepinephrine induced a dose-dependent increase in mean arterial blood pressure from 76.4 +/- 0.9 mm Hg during control to 85.6 +/- 1.5, 92.2 +/- 1.8 and 98.4 +/- 2.4 mm Hg, respectively. Ramipril significantly affected the baroreceptor set point with a decrease in mean blood pressure (72.1 +/- 1.7 vs 76.4 +/- 0.9 mm Hg, p less than 0.01) in the presence of unchanged heart rate (71.7 +/- 0.9 vs 73.6 +/- 1.5 min-1). Baroreceptor sensitivity, estimated by the slope of the delta blood pressure versus delta heart rate relation, was not affected by ACE inhibition. Also, the pressor effect of exogenous norepinephrine was unchanged by converting enzyme inhibition. The present results show that ACE inhibition with ramipril in sodium-replete healthy volunteers induces a decrease in blood pressure that is not accompanied by changes in heart rate, pressor sensitivity to exogenous norepinephrine or baroreceptor sensitivity. PMID- 3034034 TI - Sympathetic nervous activity and noradrenaline reactivity during angiotensin converting enzyme inhibition. AB - The effect of the angiotensin converting enzyme inhibitor ramipril on catecholamine disposition and noradrenaline reactivity was studied in normotensive volunteers. In the first study 5 mg of ramipril or placebo was given 3 times at 12-hour intervals in a randomized, double-blind, cross-over manner (n = 10). In the second study, ramipril 10 mg daily was given for 2 weeks (n = 6). Noradrenaline reactivity increased significantly (p less than 0.05) both in short and long-term application, while blood pressure decreased (p less than 0.01). Sulfoconjugated plasma noradrenaline decreased significantly (p less than 0.05) possibly indicating a decrease in sympathetic tone. These findings suggest that a decrease in sympathetic tone could contribute to the blood pressure-lowering effect of ramipril, whereas the increase in noradrenaline reactivity is probably a consequence of the primary change in sympathetic activity. PMID- 3034035 TI - Pharmacokinetic interaction study with ramipril and digoxin in healthy volunteers. AB - Coadministration of captopril has been shown to increase serum digoxin concentration. The effects of ramipril, a new angiotensin converting enzyme inhibitor, on serum digoxin concentration after multiple dosing were studied in 12 healthy volunteers. All subjects were receiving steady-state digoxin medication (0.5 mg daily), and ramipril (5 mg daily) was coadministered for 14 days. Serum digoxin concentration was measured repeatedly before, during and up to 1 week after ramipril coadministration at 8 a.m. (trough values) and on selected trial days at 11 a.m., 3 hours after the morning medication. Simultaneously, blood levels of ramipril and its active metabolite diacid were determined. Volunteers were followed closely for side effects and for changes in blood pressure, heart rate and electrocardiogram. Safety pharmacology included serial determination of sodium, potassium, serum glutamic oxaloacetic transaminase, creatinine and a full blood count. Mean serum digoxin concentration was not significantly influenced by ramipril coadministration with trough levels of 0.90 +/- 0.24 before, 0.93 +/- 0.38 during and 0.82 +/- 0.33 ng/ml after ramipril medication. The increase in serum digoxin concentration 3 hours after the morning dose was also not significantly affected by ramipril. Serum levels of ramipril and its diacid showed a wide range of variation. Mean serum potassium increased by 0.3 mmol/liter during ramipril coadministration with development of symptomless hyperkalemia (6.0 mmol/liter) in 1 subject. The only other side effect possibly related to ramipril was a dry cough in 1 subject. Both drugs were well tolerated. Ramipril showed no significant influence on serum digoxin levels in healthy volunteers. PMID- 3034036 TI - Pharmacokinetics and pharmacodynamics of ramipril in renal failure. AB - The pharmacokinetics and pharmacodynamics of the novel angiotensin converting enzyme (ACE) inhibitor ramipril were studied in 6 patients with a glomerular filtration rate of less than 20 ml/min/1.73 m2 of body surface area. A single oral dose of 5 mg was given and serum concentrations of the compound and its diacid, the active metabolite (ramiprilat), as well as ACE activity, blood pressure and pulse rate were monitored for 28 days. The original compound reached peak serum concentrations of 42.8 +/- 26.5 ng/ml about 1 hour after dosing and was completely eliminated from the serum after 24 hours. Ramiprilat reached peak values of 14.4 +/- 11.6 ng/ml after about 6 hours. In contrast with the parent compound, low concentrations of ramiprilat were still detected in the serum after 28 days. ACE activity decreased to approximately 5% of baseline values, remained low for the next 48 hours, then increased slowly thereafter but reached only 84.5% of initial values after 28 days. Blood pressure decreased significantly and remained low for 24 hours after dosing. The drug was well tolerated in all patients. It is concluded that a single 5 mg dose of ramipril was effective in inhibiting plasma ACE activity and lowering blood pressure in patients with renal failure. There was a slower decline in ramiprilat concentrations compared with subjects with normal renal function. PMID- 3034037 TI - Influence of renal function on the pharmacokinetics of ramipril (HOE 498). AB - The pharmacokinetics of ramipril (HOE 498) were studied after oral administration of a single 10 mg dose to 24 hypertensive patients with different degrees of renal function. The creatinine clearance ranged between 4.1 and 126 ml/min/1.73 m2 and was below 35 ml/min/1.73 m2 in 16 patients. Angiotensin converting enzyme activity and the concentrations of ramipril and its active diacid metabolite ramiprilat were measured in plasma up to 10 days after drug intake. Urine levels of ramipril, ramiprilat, their glucuronides and 2 major metabolites (a diketopiperazine and a diketopiperazine acid) were measured up to 4 days after medication. The plasma concentration-time curve of ramiprilat was polyphasic with an initial steep decline after the peak level and a subsequent very long terminal phase at low concentrations. Impaired renal function resulted in higher peak levels of ramiprilat, longer times to peak and a markedly slower decline of plasma ramiprilat levels. Hence, the duration of angiotensin converting enzyme inhibition was considerably prolonged in renal failure and depended on the severity of renal impairment. The urinary excretion of ramipril and its metabolites decreased with decreasing renal function and was linearly related to the creatinine clearance, suggesting an alternative pathway of elimination. The pattern of excretion rates of ramipril and its various metabolites was not affected by renal failure. In contrast to the marked changes in the renal elimination, no relevant differences were observed in the absorption of ramipril from the gastrointestinal tract. Systolic and diastolic blood pressure decreased in all groups. The single 10 mg dose of ramipril was well tolerated. PMID- 3034038 TI - Short- and long-term effects of ramipril in hypertension. AB - The effect of various doses of ramipril, a new converting enzyme inhibitor, was compared with that of placebo in patients with mild essential hypertension. After a single dose of 2.5 mg blood pressure was not significantly affected, despite a decrease in converting enzyme levels. Single doses of 5 or 10 mg did reduce blood pressure, although complete inhibition of the enzyme was apparent only with the higher dose. Despite partial recovery, both converting enzyme and blood pressure remained reduced for 48 hours. After 1 month of treatment with 10 mg of ramipril, renal vascular resistance had decreased and renal blood flow increased. Continued treatment for 1 year controlled blood pressure in 6 of 10 patients; in the remainder a diuretic needed to be added to maintain control of blood pressure. PMID- 3034039 TI - Effect of ramipril on 24-hour variability of blood pressure and heart rate in essential hypertension. AB - Nonrestricted blood pressure recording was performed invasively or noninvasively, using new portable devices, for a period of 24 hours in 4 patients with essential hypertension before and after 6- to 17-day treatment with ramipril at an initial dosage of 1.25 mg daily. Ramipril produced a steady decrease in blood pressure without changing heart rate. Before initiation of ramipril treatment, the blood pressure was lower during the night than during the day. This day to night difference was not affected by ramipril. In addition, analysis of the standard deviation of the mean for each time point examined during 24 hours revealed no effect of ramipril on circadian variation of blood pressure. PMID- 3034041 TI - Blood pressure response of nephrectomized subjects and patients with essential hypertension to ramipril: indirect evidence that inhibition of tissue angiotensin converting enzyme is important. AB - The kinetics of blood pressure changes and plasma angiotensin converting enzyme (ACE) inhibition in response to ramipril (HOE 498), 10 mg orally, were studied in 6 nephrectomized subjects 12 hours after ultrafiltration and in 10 patients with essential hypertension. Ramipril lowered supine and standing blood pressure in both groups, but the effect was greater in essential hypertension. The maximal blood pressure response followed the effect on plasma ACE after a lag time of 3 to 4 hours in both groups. These data provide indirect evidence that ramipril lowers blood pressure, at least in part, independently of its effect on the circulating renin-angiotensin system, possibly by acting on tissue ACE. PMID- 3034040 TI - Effect of ramipril, a new angiotensin converting enzyme inhibitor, on diurnal variations of blood pressure in essential hypertension. AB - The effect of ramipril on diurnal variations of blood pressure was studied in patients with mild to moderate essential hypertension in groups given once- (n = 18) and twice-daily (n = 21) administration with daily dosages ranging from 2.5 to 10 mg. After ramipril treatment, the blood pressure of patients in both groups was significantly reduced, and no significant differences in diurnal variation of blood pressure were observed between the 2 groups. The pulse rate did not change after administration of ramipril and no serious side effects were observed. In consideration of patient compliance, once-daily administration of ramipril seems to be optimal for the treatment of essential hypertension. PMID- 3034043 TI - Nuclear origins of brainstem reticulocortical systems in the rat. AB - Stereotaxic injections of 5% Fast Blue or 1% horseradish peroxidase-wheat germ agglutinin conjugate (HRP-WGA) were made into various cytoarchitectonic or functional regions of the cerebral cortex of anesthetized adult albino or hooded rats. Sections through the brainstems of these animals were then scrutinized for the presence of retrogradely labeled neurons. The data generated by this study indicate that at least 33 distinct nuclei or subnuclei within the brainstem reticular formation of the rat project directly to the cerebral cortex. More than half of these ascending reticulocortical systems are probably aminergic. The strongest reticulocortical projections emanate from presumed aminergic reticular cell groups located at isthmic levels: specifically, the rostral serotonin containing cell groups, as well as the noradrenergic locus coeruleus. However, relatively strong direct reticulocortical projections also originate from lower medullary cell groups which are probably catecholaminergic. Moderately strong reticulocortical projections emanate from cholinergic cell groups located at isthmic levels (the pars compacta of the pedunculopontine nucleus and the X area of Sakai). The most surprising finding in this study was that the classic isodendritic, nonaminergic central core of the brainstem gives rise to direct reticulocortical projections. The ventromedial areas of the medullary brainstem reticular formation give rise to the strongest nonaminergic ascending reticular projections, but all levels of the classic isodendritic reticular core give rise to direct reticulocortical projections. As a whole, cortically projecting reticular neurons are mostly small (10-25 microns in greatest diameter) or medium sized (26-35 microns in greatest diameter) neurons. Previous studies have shown that many of the cortically projecting reticular nuclei also project to the spinal cord, and within these nuclei, reticulocortical neurons often strongly resemble their reticulospinal counterparts with respect to details of neuronal morphology. This in turn suggests that some reticulocortical neurons may also project to spinal levels. PMID- 3034042 TI - Use of nuclear magnetic resonance imaging to show regression of hypertrophy with ramipril treatment. AB - Thirty-two patients with arterial hypertension (diastolic blood pressure greater than 95 mm Hg) were treated with ramipril for 3 months. The aim of the study was to achieve an effective decrease in blood pressure and demonstrate reliably and reproducibly that regression of left ventricular hypertrophy takes place with ramipril treatment. Nuclear magnetic resonance images and echocardiographic measurements of the left ventricle were therefore made before treatment started, 4 hours after the first dose, 14 days after the start of treatment and after 3 months of treatment. The thickness of the septum decreased from 19.57 to 15.20 mm on magnetic resonance scans and from 18.78 to 14.57 mm on echocardiograms. The values were reproduced 3 times at the same measuring point and means were calculated. The septum and posterior wall of the left ventricle were also measured at 3 different points. With negligible scatter, the values obtained were reproducible and the differences were highly significant (p = 0.001). A parallel decrease in blood pressure to levels 15% below baseline was also observed. The therapeutic aim of achieving diastolic blood pressure levels of less than or equal to 90 mm Hg was achieved in all patients. In addition to reducing the blood pressure significantly, the angiotensin converting enzyme inhibitor ramipril caused a significant regression of pathologic left ventricular hypertrophy, which was demonstrated clearly using magnetic resonance imaging and echocardiography. PMID- 3034044 TI - Phytates and the inhibitory effect of bran on iron absorption in man. AB - The cause of marked inhibitory effect of bran on absorption of dietary nonheme iron was studied in man by double-radioiron technique. Washing bran with hydrochloric acid but not with water removed inhibitory factor(s). Inhibition was almost restored by reconstituting phytate level. Removal of phytates in bran by endogenous phytase significantly increased absorption of iron. Removing, by washing with water, phosphates formed from phytates during enzymatic dephytinization led to a bran fraction with only a small remaining inhibitory effect on iron absorption. Half the iron in bran is in the form of monoferric phytate, which is well-absorbed. When potassium and magnesium phytates were added in amounts present in bran, the same inhibitory effect on iron absorption was seen. Although there appear to be other factors in bran that partly explain the inhibition, phytates are the main cause of the inhibitory effect of bran on iron absorption. PMID- 3034045 TI - Definitions and intakes of dietary fiber. PMID- 3034046 TI - Dietary fiber and carbohydrate metabolism. PMID- 3034048 TI - Fermentation in the human large intestine and the available substrates. PMID- 3034047 TI - Dietary fiber and lipids. PMID- 3034049 TI - Minerals, trace elements, and potential hazards. PMID- 3034050 TI - Dietary fiber and gastrointestinal disease. PMID- 3034051 TI - Giant neutrophils with increased peroxidase activity. Another evidence of dysgranulopoiesis in AIDS. AB - Using the automated hematologic analyzer Technicon H6000, which classifies leukocytes by their size and peroxidase activity, the authors have observed in nine patients with full-blown acquired immune deficiency syndrome (AIDS) a consistent increase in peroxidase content of circulating neutrophils. The increase in peroxidase activity was homogeneous in three patients (P less than 0.05). The most striking finding, however, was the occurrence of single abnormal neutrophils with peroxidase activity higher than the major neutrophil population (i.e., HPX [high peroxidase] cells). The importance of this phenomenon was correlated with the clinical status, higher HPX values being found in patients with more advanced disease. These instrumental observations were associated with the morphologic finding of atypical neutrophils, much larger than normal, with irregular nuclei and abundant cytoplasm filled with peroxidase-positive granulations. Such cells represent, in the authors' experience, the most common expression of dysgranulopoiesis in AIDS. PMID- 3034052 TI - Angiosarcoma of the breast following segmental mastectomy complicated by lymphedema. AB - A patient is discussed who had angiosarcoma of her lymphedematous right breast develop four years after segmental mastectomy for infiltrating ductal carcinoma. The lymphedema developed and persisted after an indolent and recurrent postoperative infection. The possibility that the second malignancy is a consequence of the chronic lymphedema, similar to the angiosarcomas of lymphedematous extremities after radical mastectomy, is cautiously entertained. This hypothesis is worthy of consideration as more breast conservation surgery is being done, with or without adjuvant radiation therapy, and accumulating evidence suggests that lymphedema of the breast is a common complication of surgery followed by radiation. PMID- 3034053 TI - Controversial histogenesis of the malignant fibrous histiocytoma. PMID- 3034054 TI - Hepatocarcinoma in a child with the Alagille syndrome. AB - A child with the Alagille syndrome of intrahepatic bile duct paucity developed hepatocarcinoma. Disabling cirrhosis had rendered this child a suitable candidate for transplantation before the discovery of carcinoma. However, the extension of the tumor outside the liver precluded the performance of this potentially life saving operation. Serial monitoring of the serum alpha-fetoprotein concentration may be of value in the early identification of tumors in pediatric candidates for transplantation. PMID- 3034055 TI - Immunohistochemical detection of ras oncogene p21 product in liver cirrhosis and hepatocellular carcinoma. AB - Expression of the ras oncogene p21 product by hepatocytes of cirrhotic liver with hepatocellular carcinoma (HCC) was examined immunohistochemically using mouse monoclonal antibody RAP-5. At the concentration of antibody used, histologically normal liver tissues were negative for p21 antigen, whereas hepatocytes of cirrhotic nodules from 80 of 92 HCC patients (87.4%), and 10 of 32 patients without HCC (59.4%) were positive. This difference was statistically significant (p less than 0.05). The incidence of p21 expression by hepatocytes was significantly higher in macronodular cirrhotic patients than in those with micronodular cirrhosis (p less than 0.05) and tended to be higher in those positive for hepatic hepatitis B virus markers than in those that were negative (p less than 0.1). All 16 patients with liver cell dysplasia, and 17 of 18 with adenomatous hyperplasias showed increased expression of p21 antigen. In HCC it was detected on tumor cells of 63 of 101 patients (62.4%). Characteristically, its expression in well-differentiated HCC was mild and uniformly diffuse, and in moderately differentiated tumors was markedly heterogeneous in both intensity and distribution, whereas no expression was observed in cells of poorly differentiated HCC. These observations suggest that elevated ras p21 antigen expression is important in the development of both cirrhotic nodules and HCC, but that after tumor development, its sustained elevation is no longer necessary for cell proliferation and progression through the grades of anaplasia. PMID- 3034056 TI - Antibodies to vesicular stomatitis New Jersey type virus in wild and domestic sentinel swine. AB - Wild sentinel swine on Ossabaw Island, Chatham County, Georgia, were serially bled and tested for vesicular stomatitis New Jersey type virus neutralizing antibody to determine the intensity, distribution, and progression of annual viral activity. From March through September, 1984 and 1985, 112 and 226 juvenile (less than 8 months) swine, respectively, were sampled. Seroconversions initially were detected on May 7, 1984 and May 18, 1985. Incidence of seroconversion in wild swine reached 32% during 1984 and 26% during 1985. Viral activity as determined by seroconversion results occurred earliest and was greatest on the southern half of Ossabaw Island. Domestic swine were housed in four pens under controlled conditions to document arthropod transmission of vesicular stomatitis virus. Twelve swine, three in each pen, were serially bled from April through September of both years. Seroconversion occurred during May 16-23, 1984 and May 15-22, 1985. Results varied among pen locations but were consistent between years. Clinical disease was not seen in any wild or domestic swine during either year. PMID- 3034057 TI - Mechanisms of action of and resistance to ciprofloxacin. AB - Ciprofloxacin and other newer quinolone antimicrobial agents exhibit increased potency and decreased frequency of spontaneous bacterial resistance in comparison with older analogues such as nalidixic acid. New and published observations on the mechanisms of action of and resistance to ciprofloxacin in Escherichia coli are presented and discussed. Genetic and biochemical studies have identified the A subunit of the essential bacterial enzyme DNA gyrase as a target of ciprofloxacin and other quinolones. For a series of quinolones, inhibition of purified DNA gyrase correlated with antibacterial activity. The bactericidal activity of ciprofloxacin and ofloxacin is, in contrast to that of certain other quinolones, somewhat less affected by rifampin and cell starvation, suggesting the existence of a site of drug action in addition to DNA gyrase. The frequency of selection of spontaneous single-step resistance mutants of E. coli was more than 100-fold lower with ciprofloxacin than with nalidixic acid. Strains highly resistant to ciprofloxacin could, nevertheless, be selected by serial passage on drug-containing agar. Two mutations contributing to this high level of resistance were analyzed. One, designated cfxA, conferred a 16-fold increase in drug resistance and mapped in a location consistent with a gyrA mutation; similar increases in resistance to ciprofloxacin were seen with gyrA mutations selected for resistance to other quinolones. The other mutation, cfxB, conferred pleiotropic resistance to ciprofloxacin, tetracycline, and chloramphenicol and appeared to be an allele of the multiple antibiotic resistance gene marA. The mutation cfxB was associated with a decreased amount of porin outer membrane protein OmpF, suggesting that drug permeation may occur in part through this channel. In summary, the A subunit of DNA gyrase is a target of ciprofloxacin and other quinolones. Ciprofloxacin resistance appears to occur both by mutation in this target and by alteration of drug permeation through the outer membrane of the cell. PMID- 3034058 TI - Stromal reactions to squamous cell carcinoma of the cervix. AB - The characteristics of the stromal reaction to invasion by cancer were studied in 23 cases of cervical squamous cell carcinoma (stage Ib to IIb) and 86 control cases of benign disease of the uterus. Tissues taken from sites adjacent to tumor or from comparable sites in the nonmalignant uteri were analyzed for number of round cells, density of collagen and reticular fibers, and collagenase activity. The results were correlated with the depth of invasion and the histologic appearances of the tissues adjoining the cancer. Plasma cells and reticular fibers were most numerous in patients with slight invasion and no direct contact between the malignant and benign tissues (C type in CPL classification of Imai). Glycosaminoglycans (mainly hyaluronic acid and chondroitin sulfate) were present in the greatest amounts in C type stroma. The prognosis showed no correlation with either collagen content or collagenase activity but it was a better prognosis correlated with the collagen/collagenase ratio. It is concluded that increased numbers of plasma cells and reticular fibers and a high collagen/collagenase ratio are among the mechanisms protective against the invasiveness of cervical carcinoma. PMID- 3034059 TI - Effects of enprofylline, a new xanthine derivate, on human pregnant myometrium. AB - The recently developed xanthine derivate, enprofylline, was studied in an in vitro system with human pregnant myometrium, in particular its effect on spontaneous myometrial contraction, cyclic adenosine monophosphate content, cyclic adenosine monophosphate protein kinase, and cyclic adenosine monophosphate and cyclic guanosine monophosphate-dependent phosphodiesterase activity. Enprofylline was shown to be a rather potent smooth muscle relaxant [inhibitory concentration that decreases response by 50% (IC50) = 3 X 10(-5) mol/L] and an almost equally potent cyclic adenosine monophosphate-dependent phosphodiesterase inhibitor (IC50 = 10(-4) mol/L), whereas it was a less potent cyclic guanosine monophosphate-dependent phosphodiesterase inhibitor. Enzyme kinetic studies revealed that enprofylline is a competitive cyclic adenosine monophosphate phosphodiesterase inhibitor. Enprofylline increased the cyclic adenosine monophosphate content in a dose-dependent way with subsequent increased activity of cyclic adenosine monophosphate-dependent protein kinase. It is suggested that enprofylline relaxes myometrial smooth muscles and that this is at least partly the result of interference with the cyclic adenosine monophosphate system. PMID- 3034060 TI - Delayed amplification of cytomegalovirus infection in the placenta and maternal tissues during late gestation. AB - To stimulate first-trimester human cytomegalovirus infection, pregnant guinea pigs were inoculated with a low dose of guinea pig cytomegalovirus during the first trimester of pregnancy. Maternal viremia, which was cleared by 2 weeks after inoculation, was found to reappear near the time of delivery in one third of the animals tested. The virus, first detected in the placenta during initial maternal viremia, replicated after the time when maternal blood was cleared of virus, although high titers of maternal serum antibodies were present. In the last week of gestation (43 to 48 days after inoculation), the virus was detected in 95% of placentas and was present at high titers. Fetal infection first appeared on day 25 after inoculation and reached an incidence of 37% in the last week of gestation despite the presence of fetal antibody. These results suggest that the placenta may amplify cytomegalovirus infection late in human gestation, even after low-dose infection in the first trimester. PMID- 3034061 TI - Effect of estrogen on dehydroepiandrosterone formation by baboon fetal adrenal cells in vitro. AB - The present study determined if estrogen modulates the responsivity of the adrenal gland of the baboon fetus to tropic hormones such as adrenocorticotropic hormone and prolactin. Adrenal glands were obtained from seven baboon fetuses at midgestation (days 100 to 105). Adrenal cells were dispersed in medium 199 with 0.2% collagenase for 10 minutes at 37 degrees C. Approximately 10(5) cells/4.0 ml of medium 199 were incubated for 24 hours at 37 degrees C with 10 nmol of adrenocorticotropic hormone or 10 nmol of ovine prolactin in the presence or absence of 10(-5) or 10(-6) mol/L of estradiol. The major steroid formed and secreted into the medium was dehydroepiandrosterone. Mean +/- standard error basal formation of dehydroepiandrosterone was 176 +/- 64 ng/10(5) cells/24 hours. Dehydroepiandrosterone formation was increased (p less than 0.05) 3.5-fold and five-fold by adrenocorticotropic hormone and prolactin, respectively. Estradiol at 10(-5) mol/L prevented the response in dehydroepiandrosterone obtained with adrenocorticotropic hormone alone. Estradiol alone had no effect on dehydroepiandrosterone. The results suggest that estrogen modulates the regulatory effects of adrenocorticotropic hormone on dehydroepiandrosterone formation by the adrenal gland of the baboon fetus. PMID- 3034062 TI - Efficacy of transdermal estradiol. AB - Several side effects and risks associated with estrogen replacement therapy are known to stem from the hormone's impact on the liver. With oral administration, the enhanced action at hepatic, as compared with nonhepatic, sites is presumably related to the so-called first-pass effect. Attempts have been made to avoid this action by administering estrogen nonorally, but heightened hepatic effects (comparable with those of other preparations) have nonetheless been seen with both ethinyl estradiol and conjugated equine estrogens given vaginally. We conducted a series of investigations aimed at evaluating the effects of estradiol delivered via a transdermal patch. In a 50-patient study of transcutaneous estradiol (25, 50, 100, or 200 micrograms/day) versus placebo, a dose-dependent beneficial effect on objectively measured hot flashes was demonstrated. A second study was designed to compare the effects of these doses with those of 0.625 and 1.25 mg conjugated equine estrogen administered orally. Effects on nonhepatic markers were similar for the 50 micrograms patch and 0.625 mg tablet, as well as for the 100 micrograms patch and 1.25 mg tablet. None of the doses of transdermal estradiol exerted any measurable action on hepatic markers of estrogen action, whereas both doses of conjugated equine estrogen demonstrated actions on both hepatic protein and lipid synthesis. Our data clearly show that the transdermal administration of estradiol circumvents the enhanced hepatic actions of the hormone. Possible explanations for these results are presented. PMID- 3034063 TI - Epstein-Barr viral antibodies in multifocal choroiditis and panuveitis. AB - A syndrome of multifocal choroiditis, pigment epithelial disturbance, and inflammatory vitreous cells was found to be associated with Epstein-Barr virus specific antibodies. The ten patients in this series had positive viral capsid antigen IgM or Epstein-Barr early antigen antibody titers. Patients with this syndrome were generally healthy and had no history of a clinical episode of infectious mononucleosis. No patient from the control group with other ocular diseases had positive viral capsid antigen IgM or early antibody titers. All patients in the study group and most of the control patients had viral capsid antigen IgG and Epstein-Barr nuclear antigen antibodies, indicating a previous exposure to the virus as expected in an adult population. PMID- 3034064 TI - Association of human papillomavirus type 16 with neoplastic lesions of the vulva and other genital sites by in situ hybridization. AB - The authors examined paraffin sections from 85 genital tract tissues from 49 cases for the presence of human papillomavirus (HPV) Types 6/11, 16, and 18 by stringent in situ hybridization using 35S-labeled viral DNA probes, and for viral capsid antigen by the immunoperoxidase test. The cases, selected mostly on the basis of vulvar pathology, were distributed as follows: early neoplasia (Group I, 6 cases); early neoplasia with viral cytopathic effect (CE) (Group II, 24 cases); and papillomavirus infection (PVI) (Group III, 19 cases). Available tissues from all affected sites were examined when the disease was multicentric. One or more viral DNAs were identified in 58% of 77 tissues from Groups II and III and in 2 of 8 tissues from Group I. HPV-6/11, HPV-16 and HPV-18 DNAs were detected, respectively, in 25, 24, and 2 tissues; 3 tissues were infected simultaneously with either two or three viruses. Viral DNA was identified at more than one site in 14 of 30 DNA-positive patients; in 10 of these, a single type was detected at all sites in the same patient. The viral DNA was localized mostly in areas showing viral cytopathology. The presence of HPV-16 correlated with neoplasia. HPV-16 DNA was identified in the 2 virus-positive tissues showing neoplasia, in 17 of 20 (85%) of the DNA-positive tissues showing neoplasia with CE, and in 5 of 25 (20%) of the DNA-positive tissues showing PVI. Conversely, HPV-6/11 was found in 25% of the DNA-positive tissues showing neoplasia with CE and in 80% of the cases of PVI. An HPV genome was identified in neoplastic cells in 14 instances; in all but 1 case, the genome was HPV-16. The association of HPV-16 with neoplasia was seen for both vulvar and cervical lesions. Viral antigen was detected in 83% of lesions associated with HPV 6/11 and in 62% of lesions associated with HPV-16. PMID- 3034065 TI - Lung injury mediated by antibodies to endothelium. II. Study of the effect of repeated antigen-antibody interactions in rabbits tolerant to heterologous antibody. AB - The effect of repeated interactions of antibodies with cell surface antigens have been examined in in vitro, but not in in vivo systems. In this study are described the results of multiple antibody-cell surface antigen interactions in vivo. Rabbits were given repeated intravenous injections of goat antibodies to angiotensin converting enzyme (ACE), an antigen expressed on the surface of lung endothelial cells. For prevention of anaphylactic reactions, which would have been induced by multiple injections of heterologous immune or nonimmune IgG, the rabbits were made neonatally tolerant to goat IgG. Divalent immune IgG given daily for 21 days induced chronic antigenic modulation (antigen disappearance) with resistance to antibody-mediated inflammatory lesions. The rabbits, however, developed degenerative changes of alveolar endothelial and epithelial cells. Administration of immune IgG every other day for 43 days allowed partial reexpression of ACE and was associated with intravascular, but not interstitial, inflammatory changes. In contrast, repeated administration of monovalent immune Fab did not induce antigenic modulation but caused severe, lethal, interstitial pneumonitis. Thus, in this experimental model the development of acute interstitial inflammatory changes correlates with persistence of antigen and is abrogated by disappearance of antigen induced by divalent antibodies. Further, repeated endothelial antigen antibody interactions fail to induce chronic inflammatory or sclerosing lung lesions. PMID- 3034066 TI - Murine cytomegalovirus infection of cultured mouse embryos. AB - Isolated mouse whole embryos of 7.5 days' gestation were infected with murine cytomegalovirus (MCMV) and cultured in pure rat serum. Although the MCMV infection had little effect on the survival and development of the embryos during 3 days of cultivation, immunohistochemical analysis of their serial sections using monoclonal antibody showed MCMV-infected cells in various portions of the embryos. This monoclonal antibody, when tested with the use of infected cultured mouse fibroblasts, reacted with nuclear antigen within 2 hours after infection and also reacted with nuclear inclusions in the late phase of infection. The viral antigen-positive cells detected by the monoclonal antibody were present in almost all of the ectoplacental cone and the yolk sac and in about 82% of the embryos. In the embryos, antigen-positive cells were frequently observed in the epithelium of the digestive tracts, endothelial cells of the blood vessels, and the mesodermal cells. In some of the embryos, viral antigen-positive cells were clearly observed in a small percentage of the blood cells. These findings indicate that blood cells, in addition to cell migration during embryogenesis, may play an important role in transmission of infectious virus into the embryos. Mouse whole embryo culture infected with MCMV can provide a model for the study of cellular tropism related to congenital infection by cytomegalovirus. PMID- 3034069 TI - Aldosterone regulation of Na+ transport and Na+-K+-ATPase in A6 cells: role of growth conditions. AB - The effects of aldosterone on transepithelial sodium transport (measured by the short-circuit current (SCC) and on Na+-K+-adenosine triphosphatase (ATPase) biogenesis have been studied in A6 kidney cells grown on collagen-coated filters in two different media. In medium A, base-line SCCA was close to zero but transmural electrical resistance (RA) was high. Aldosterone (100 nM, t24h) drastically increased SCCA and RA, but only after a 4-h latent period. In medium B, base-line SCCB and RB were significantly higher than in medium A. Aldosterone significantly enhanced SCCB and to a lesser extent RB after a much shorter latent period (approximately 45 min) than in medium A. In medium A, aldosterone elicited a fourfold increase in the relative rate of synthesis of alpha- and beta-subunits of Na+-K+-ATPase. A twofold increase was already observed within the observed latent period. This time course suggests that de novo synthesis of sodium pumps might be one of the critical factors underlying the increase in sodium transport in this growth medium. In medium B, aldosterone elicited a two- to fourfold increase in the relative rate of synthesis of the alpha- and beta-subunits of Na+ K+-ATPase that paralleled SCCB. Thus de novo synthesis of Na+-K+-ATPase is clearly not a prerequisite for the early mineralocorticoid response (t90 min - t180 min), but still could be part of the late mineralocorticoid response (t3 h - t24 h). In both media, the immunochemical cellular pool of Na+-K+-ATPase was apparently not modulated by aldosterone for up to 48 h of incubation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3034067 TI - Phospholipid base exchange activity in the leukocyte membranes of patients with inflammatory disorders. AB - Phospholipid base exchange and cholinephosphotransferase (CPT) and ethanolaminephosphotransferase (EPT) activities were assessed in the membranes of neutrophils or lymphocytes from patients with various inflammatory disorders. Ethanolamine exchange activity was significantly enhanced in both neutrophils and lymphocytes from patients with active Behcet's disease, active systemic lupus erythematosus (SLE), and severe bacterial infections and slightly enhanced in those from patients with active rheumatoid arthritis (RA), compared with healthy controls. No abnormal findings were found in CPT, EPT, or serine or choline base exchange activities in the leukocytes from any of the diseased groups tested or in the ethanolamine exchange activity of patients with severe viral infections and inactive SLE, RA, and Behcet's disease. The authors have recently demonstrated the enhancement of transmethylation and phospholipase A2 activity in human leukocyte membranes at the height of inflammatory disease states, as well as the activation of leukocyte ethanolamine exchange by bioactive stimulants. These data postulate that phosphatidylethanolamine synthesis by the base exchange reaction may be the precursor of transmethylation and its subsequent activation of phospholipase A2, leading to the induction of arachidonic acid cascade. PMID- 3034068 TI - Coxsackievirus B3-induced myocarditis. Autoimmunity is L3T4+ T helper cell and IL 2 independent in BALB/c mice. AB - Male BALB/c mice inoculated with 6 X 10(4) plaque-forming units (pfu) coxsackievirus, group B, type 3 (CVB3), develop myocarditis within 7 days. Two cytolytic T lymphocyte (CTL) populations arise in infected animals. One population belongs to the Lyt 2+ T (cytolytic/suppressor) lymphocyte subset and reacts specifically with uninfected heart cells (autoreactive CTLs, ACTLs), whereas the other belongs to the L3T4+ T (helper) lymphocyte subset and reacts with infected targets (virus-specific CTLs, VSCTLs). Although both immune T lymphocyte populations can induce cardiac inflammation in vivo, ACTLs predominantly cause tissue injury. VSCTL generation can be inhibited by either anti-Tac (antibody to the interleukin 2 [IL-2] receptor) or anti-Iad but not by anti-IAk, indicating that this response is probably both IL-2-dependent and Class II (major histocompatibility complex [MHC] antigen restricted. ACTL generation is independent of IL-2, because neither anti-Tac or cyclosporin A inhibit this response. PMID- 3034070 TI - Characterization of Na+-H+ antiport in type II alveolar epithelial cells. AB - The presence of a Na+-H+ exchange pathway in the plasma membrane of type II alveolar epithelial cells was explored using the pH-sensitive fluorescent probe 2,7-biscarboxyethyl-5,6-carboxyfluorescein (BCECF) to monitor changes in cytosolic pH. Freshly prepared pneumocytes suspended in medium at pH 7.4 had an intracellular pH of 7.07 +/- 0.07. Acid-loaded cells equilibrated in sodium-free buffer showed rapid cytoplasmic alkalinization when exposed to sodium. This response to sodium was inhibited greater than 90% by 10(-4) M amiloride. The presence of the K+ ionophore, valinomycin, had no effect on the rate of Na+ dependent alkalinization, indicating the electroneutrality of the system. Li+ partially supported the alkalinization process, but other monovalent cations, notably K+, Rb+, and Cs+, were without effect. Kinetic analysis for Na+ at the external binding site yielded KNat (dissociation constant) = 62 +/- 3 mM. Hill equation analysis of the data derived a Hill coefficient (n) = 1.2 +/- 0.1 for Na+, consistent with a 1:1 stoichiometry for Na+ and H+ for the transporter. The Ki for amiloride inhibition of proton efflux at the external locus was 0.45 microM. These findings define the transport pathway as Na+-H+ antiport, with kinetic parameters somewhat similar to those described for other cell types. Antiport activity was detected at intracellular pH (pHi) values of 6.8 or below, with no activity observed at pHi 7.0-7.2. It is suggested that Na+-H+ exchange is a major mechanism whereby pneumocytes recover from an acid load and that this transport pathway may play an important role in vectorial reabsorption of Na+ from the alveolar air spaces. PMID- 3034071 TI - Evidence for lack of a role of cGMP in effect of alpha-hANP on aldosterone inhibition. AB - To investigate the role of guanosine 3',5'-cyclic monophosphate (cGMP) in the inhibitory effect on aldosterone production of alpha-human atrial natriuretic polypeptide (alpha-hANP) we first compared the effects of the peptide with those of sodium nitroprusside (SNP) on the production of aldosterone and cGMP in dispersed adrenal capsular cells of rats, second, examined the effects of derivatives of cGMP on the production of aldosterone, and, third, studied the influence of potassium on the effects of alpha-hANP on the production of aldosterone and cGMP. alpha-hANP at concentrations of 3 X 10(-8) to 3 X 10(-7) M decreased the production of aldosterone in a dose-dependent manner, while markedly increasing the production of cGMP. On the other hand, although SNP at concentrations of 10(-5) to 10(-3) M increased the production of cGMP in a dose dependent manner, it caused no significant changes in the production of aldosterone. Neither dibutyryl cGMP nor 8-bromo-cGMP affected the production of aldosterone in the adrenal cells. Although the aldosterone-inhibitory effect of alpha-hANP was lost in the potassium-free medium, the cGMP-stimulatory effect of the peptide was not altered by adding potassium to the incubation medium at concentrations of 0-5 meq/l. These results suggest that cGMP plays a minor role in the inhibitory effect of alpha-hANP on the production of aldosterone and that the production of cGMP stimulated by the peptide is not directly involved in the decrease in aldosterone production in adrenal capsular cells of rats. PMID- 3034072 TI - Altered hepatic vasopressin and alpha 1-adrenergic receptors after chronic endotoxin infusion. AB - Sepsis and septic shock are complicated by a number of hemodynamic and metabolic aberrations. These include catecholamine refractoriness and altered glucose metabolism. Recently, a nonshock rat model of continuous endotoxin infusion via an implanted osmotic pump was developed that reproduces some of the metabolic and cardiovascular findings of human sepsis. By using this model, we have found a decreased number of hepatic plasma membrane alpha 1-adrenergic and [Arg8]vasopressin receptors in rats continuously infused with endotoxin. There was a significant decrease in [3H]prazosin (35 +/- 7%) and [3H] [Arg8]vasopressin (43 +/- 8%) receptors after 30 h of continuous endotoxin infusion with no change in affinity. The ability of norepinephrine to form the high-affinity complex with alpha 1-adrenergic receptors was not altered after chronic endotoxin infusion. The results are consistent with the concept that alterations in receptor number might underlie certain of the metabolic consequences of chronic sepsis. PMID- 3034073 TI - Renal tubular secretion of the alkanesulfonate 2,3-dimercapto-1-propanesulfonate. AB - The in vivo tubular secretion and metabolism of 2,3-dimercapto-1-propanesulfonate (DMPS) was examined in the chicken by use of the Sperber technique. Infusion of DMPS into the renal portal circulation of the chicken at rates equal to or less than 7.5 mumol X min-1 X kg body wt-1, resulted in a tubular excretion ratio of 0.5 for DMPS with 90% of the infused DMPS excreted in the urine unchanged. Renal tubular secretion accounted for approximately 90% of the total DMPS excreted into the urine during the infusion of DMPS at a rate of 0.75 mumol X min-1 X kg-1. The secretion of DMPS was saturable and was inhibited by p-aminohippurate (PAH), probenecid, and heptanesulfonate. Taurine (2-aminoethanesulfonate), isethionate (2-hydroxyethanesulfonate), and 2-mercaptoethanesulfonate had no effect on the secretion of DMPS or PAH. Renal tubular secretion may explain several pharmacological characteristics previously reported for DMPS, including the selective removal of heavy metals from the kidney. PMID- 3034074 TI - Superficial nephron responses to peritubular capillary infusions of angiotensins I and II. AB - Proximal tubular reabsorption, stop-flow pressure (SFP), and single nephron glomerular filtration rate (SNGFR) were measured in the absence of and during infusion of an isotonic saline solution containing either angiotensin I (ANG I; 10(-6) to 10(-5) M) or angiotensin II (ANG II; 10(-9) to 10(-7) M) into an adjacent peritubular capillary at a rate of 20 nl/min. Dilution of the infused ANG I and ANG II occurred in the peritubular capillary blood and as the peptides diffused into the interstitium. Infusion of either 10(-7) M ANG II or 10(-5) M ANG I increased proximal fractional fluid reabsorption (FRH2O) and decreased both SFP and SNGFR. There were no significant changes in FRH2O or SNGFR during infusion of 10(-5) M ANG I when the converting enzyme inhibitor enalaprilat (MK 422, 10(-3) M) was added to the infusate. Similarly, peritubular infusion at lower concentrations of either ANG II (10(-9) or 10(-8) M) or ANG I (10(-6) M) did not alter FRH2O, SFP, or SNGFR. These data indicate that conversion of ANG I to ANG II can occur in the peritubular capillary or interstitial environment and that increases above the normal endogenous levels in the postglomerular interstitial ANG II concentration can enhance proximal tubular reabsorption and increase preglomerular resistance and thereby reduce SNGFR. PMID- 3034075 TI - Na+-K+-ATPase activity in medullary thick ascending limb during short-term anoxia. AB - Na+-K+-ATPase activity was measured in a suspension of rabbit medullary thick ascending limb tubules under oxygenated and anoxic conditions. Oxygenated, K depleted tubules rapidly take up added extracellular potassium accompanied by a simultaneous increase in oxygen consumption. The ATP/O2 ratio was 12.5 +/- 0.7, suggesting a tight coupling between oxidative metabolism (6 ATP/O2) and Na+-K+ ATPase activity (2 K/ATP). On reaching anoxia, the tubules released potassium into the medium, but this rate was accelerated by the addition of ouabain, which indicated that the Na+-K+-ATPase was still operative in anoxia. Because 10 min of anoxia led to only a 15.7% decline in potassium content, a new steady state of potassium uptake and leakage must be reached during anoxia. Anaerobic metabolism maintained 73% of cellular ATP during 10 min of anoxia. Exposure of anoxic tubules to iodoacetate produced a 57% decline in ATP levels and a 33% decline in potassium content, which indicated that glycolysis is an important pathway in supplying energy during anaerobiosis. PMID- 3034076 TI - Cl-/HCO-3 antiporter in red cell ghosts: a kinetic assessment with fluorescent probes. AB - The pH sensitive fluorescent probe acridine orange and membrane potential sensitive fluorescent probe acridine orange and membrane potential-sensitive fluorescent probe 3,3'-dipropylthiadicarbocyanine iodide were used to evaluate the Cl-/HCO-3 antiporter and proton and potassium conductances, respectively, in human red blood cell ghosts. Acidic, chloride-loaded ghosts alkalinized rapidly in pH 8.5 chloride-free media. Alkalinization could not be ascribed to conductive proton efflux with either depolarizing potassium influx or chloride efflux. Alkalinization was consequent to flux on the Cl-/HCO-3 antiporter: this process displayed saturation kinetics, competitive inhibition by external chloride, and inhibition by 4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonic acid. The mean Kms for internal chloride, external bicarbonate, and external chloride were 2.19, 0.24, and 0.44 mM, respectively. These studies confirm both the asymmetry of this carrier and the high affinity for external HCO-3; however, the affinities for internal and external chloride are significantly greater than prior estimates. The Km for internal chloride (2.19 mM) was considerably lower than levels previously reported (20-65 mM) unless external (trans) chloride was raised above 2 mM. The present studies thus demonstrate and emphasize the critical importance of trans substrate concentration in assessing the kinetics of a carrier whose mobility is faster in the loaded than in the unloaded state. PMID- 3034077 TI - Electrophysiological studies of primary cultures of rabbit distal tubule cells. AB - Primary monolayers grown from F1 band of a Percoll gradient centrifugation ("distal" monolayers) were studied, after confluency, 6-14 days after seeding. Transmission and scanning electron microscopy revealed that two cell types, resembling principal cells of the rabbit cortical collecting tubule (CCT) and intercalated cells of either CCT or connecting tubule, constitute approximately 96% of the monolayer. About two-thirds of the intercalated cells fluoresced when treated with fluorescent peanut lectin. Indirect specific immunocytofluorescent staining revealed fluorescence in 96% of the cells, confirming that the monolayers were derived from CCT or connecting tubule cells. Exposure of monolayers to vasopressin or isoproterenol increases adenosine 3',5'-cyclic monophosphate (cAMP) content in the cells and bathing medium, whereas parathyroid hormone was ineffective. Electrophysiological studies revealed a transepithelial voltage (VT) of -11 +/- 2 mV, and basolateral membrane voltage (Vb) of -77 +/- 5 mV (n = 20). The transepithelial electrical resistance (RT) was 1,870 +/- 250 omega X cm2 (n = 13). In three out of six monolayers, amiloride (10(-5) M) applied to the apical side produced an increase in apical membrane voltage (Va) from -71 +/- 1 to -89 +/- 9 mV) and a decrease in VT (from -10 +/- 1 to -2 +/- 1 mV). The RT did not change during amiloride exposure. Exposure of the apical membrane to 140 mM K+-depolarized Va from -67 +/- 7 to -39 +/- 11 mV (P less than 0.002) and hyperpolarized VT from -7 +/- 2 to -15 +/- 3 mV (P less than 0.005). Exposure to high K+ from the basolateral side depolarized Vb from -76 +/- 11 to 43 +/- 10 mV (P less than 0.001) and depolarized VT from -9 +/- to 8 +/- 5 mV (P less than 0.001). This preparation is suitable to study basic aspects of epithelial transport by electrophysiological methods and other techniques. The findings are consistent with several of the known properties of cortical collecting tubules from rabbits studied by the isolated perfused tubule technique. PMID- 3034078 TI - Enhanced glomerular filtration and Na+-K+-ATPase with furosemide administration. AB - To evaluate the effect of furosemide on kidney function, glomerular filtration rate (GFR), urinary Na excretion (UNaV), Na reabsorption (NAR), and Na+-K+-ATPase in isolated nephron segments were measured in 1) rats treated with furosemide 10 mg X 100 g-1 X 24 h-1 ip for 7 days, and 2) rats receiving an oral Na load for 12 days. In furosemide-treated rats, GFR rose from 0.61 +/- 0.03 (mean +/- SD) to 0.83 +/- 0.06 ml/min (P less than 0.01), UNaV rose from 904 +/- 71 to 1,402 +/- 85 mueq/day (P less than 0.001), and net NAR rose from 87.5 +/- 3.7 to 116.7 +/- 9.0 mueq/min (P less than 0.01). Na+-K+-ATPase remained unchanged in the proximal convoluted tubule (PCT), proximal straight tubule (PST), cortical thick ascending limb of Henle's loop (cTALH), and medullary thick ascending limb of Henle's loop (mTALH), but was increased in the distal convoluted tubule (DCT) and in cortical collecting duct (CCD) from 48.5 +/- 1.2 to 75.3 +/- 0.7 (P less than 0.001) and from 18.6 +/- 0.7 to 27.1 +/- 2.7 (P less than 0.02) X 10(-11) mol X mm-1 X min 1, respectively. In Na-loaded rats GFR rose from 0.61 +/- 0.04 to 0.86 +/- 0.03 ml/min (P less than 0.001), UNaV rose from 1,064 +/- 118 to 18,532 +/- 2,045 mueq/day (P less than 0.001), net NAR from 88.1 +/- 3.0 to 107.8 +/- 3.9 mueq/min and Na-K-ATPase in the mTALH rose from 40.3 +/- 1.4 to 56.2 +/- 2.11 (P less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3034079 TI - Two classes of ouabain binding sites in ferret heart and two forms of Na+-K+ ATPase. AB - In partially purified Na+-K+-adenosinetriphosphatase (ATPase) obtained from ferret heart, ouabain produced a monophasic inhibition curve; however, the curve spanned over 5 logarithmic units, indicating the presence of more than one classes of enzyme. [3H]ouabain binding studies revealed high-and low-affinity binding sites in approximately equal abundance, with apparent dissociation constants of 10 and 230 nM, respectively. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of phosphoenzyme formed from [gamma-32P]ATP showed two distinct K+-sensitive bands of approximately 100,000 molecular weight. Phosphoenzyme formation from the high-molecular-weight alpha(+) form was selectively inhibited by N-ethylmaleimide. Ouabain caused a 50% inhibition of phosphorylation of the alpha(+) form at 40 nM and the lower-molecular-weight alpha form at 300 nM. In papillary muscle preparations, 1-30 nM ouabain produced a modest positive inotropic effect that reached an apparent plateau at 30 nM. Further increases in ouabain concentrations, however, produced additional and prominent inotropic effects at 0.1-10 microM. These results indicate for the first time in cardiac muscle that the high- and low-affinity ouabain binding sites are associated with the alpha(+) and alpha forms of the Na+-K+-ATPase, respectively, and that binding of ouabain to either of these sites causes enzyme inhibition and the positive inotropic effect. PMID- 3034080 TI - Mechanism of alpha-adrenergic excitation in bovine lymphatic smooth muscle. AB - Measurements were made, using the double sucrose-gap technique, of electrical and mechanical responses of bovine lymphatic smooth muscle to constant current pulses. After beta-blockade with 10(-6) M propranolol, stimulation of the alpha receptors with norepinephrine (5 X 10(-6) M) depolarized the membrane and decreased membrane conductance. The depolarization and decrease in membrane conductance persisted in Li+- and choline-substituted low-Na+ solution, in methanesulfonate-substituted low-Cl- solution, and in Ca2+- free solution containing 1 mM ethyleneglycol-bis(beta-aminoethylether)-N, N'-tetraacetic acid. Tetraethylammonium (10 mM) did not itself affect membrane resistance nor did it block the increase in resistance due to norepinephrine. In contrast, cesium (10 mM) increased membrane resistance and prevented norepinephrine from increasing this further. As well as these effects on membrane resistance, norepinephrine (5 X 10(-6) M) increased the duration of the action potential, and this was accompanied by an increased force of contraction. Tetraethylammonium prolonged the action-potential plateau and potentiated norepinephrine's effect. These results suggest that norepinephrine is likely to increase the efficiency of lymphatic pumping due to both its positive inotropic effect and the improved safety margin for propagation resulting from the increase in membrane resistance. The latter effect may be due to the suppression of an outward K+ current. PMID- 3034081 TI - Proton-stimulated Cl-HCO3 antiport by basolateral membrane vesicles of lobster hepatopancreas. AB - Purified epithelial basolateral membrane vesicles were prepared from lobster hepatopancreas by sorbitol gradient centrifugation. Na+-K+ adenosinetriphosphatase, alkaline phosphatase, and cytochrome-c oxidase enzyme activities in the final membrane preparation were enriched 9.6-, 1.4-, and 0.4 fold, respectively, compared with their activities in the original tissue homogenate. Vesicle osmotic reactivity was demonstrated using 60-min equilibrium 36Cl uptake experiments at a variety of transmembrane osmotic gradients. 36Cl uptake into vesicles preloaded with HCO3 was significantly greater than into vesicles lacking HCO3. This exchange process was stimulated by a transmembrane proton gradient (internal pH greater than external pH). Proton-gradient-dependent Cl-HCO3 exchange was potential sensitive and stimulated by an electrically negative vesicle interior. 36Cl influx (4-s exposures) into HCO3-loaded vesicles occurred by the combination of 4-acetamido-4'-isothiocyanatostilbene-2,2' disulfonic acid sensitive, carrier-mediated transfer and "apparent diffusion." 36Cl influx was a hyperbolic function of both internal [HCO3] and internal [Cl]. The two internal anions displayed a 100-fold difference in apparent affinity constants with HCO3 being strongly preferred. 36Cl influx was stimulated more by preloaded monovalent than by divalent anions. Na was an inhibitor of proton dependent anion antiport, whereas K had no effect. A model for HCl-HCO3 antiport is suggested that employs combined transmembrane concentration gradients of Cl and HCO3 to power anion exchange and transfer protons against a concentration gradient. PMID- 3034082 TI - Effect of immobilization on collagen synthesis in rat skeletal muscles. AB - The activities of prolyl 4-hydroxylase (PH) and galactosylhydroxylysyl glucosyltransferase (GGT), both enzymes of collagen biosynthesis, and the concentration of hydroxyproline (Hyp) were measured in the soleus, gastrocnemius, and tibialis anterior muscles of rats after cast immobilization in the middle position for 1 or 3 wk. The specific activity of PH decreased by 54 and 70-75% (P less than 0.001) in the soleus muscle after 1 and 3 wk, respectively, the corresponding decreases in GGT activity were 43 and 47% (P less than 0.001). A less pronounced decrease in the activities of these enzymes was observed in gastrocnemius and tibialis anterior muscles. The Hyp concentration in the soleus increased during the first week of immobilization but began to decrease thereafter, and the total muscular Hyp content was reduced after immobilization for 3 wk. The results suggest a marked inhibition of muscular collagen synthesis during immobilization. Electrical stimulation of the sciatic nerve partially prevented this disuse atrophy and the decreases in PH, GGT, and Hyp in the tibialis anterior muscle but not in the gastrocnemius or soleus muscles. PMID- 3034083 TI - Beta-endorphin, ACTH, and cortisol response to hemorrhage in conscious pigs. AB - Some of the interrelations of neuroendocrine changes associated with hypovolemia were investigated in a model simulating an arterial hemorrhage. beta-Endorphin, adrenocorticotropin hormone (ACTH), and cortisol levels were measured by radioimmunoassay before, during, and after controlled bleeding of conscious splenectomized pigs. All animals showed significant (P less than 0.05) increases in the three neuroendocrine substances during hemorrhage. beta-Endorphin values initially were 55 +/- 7 pg/ml (+/- SE) and rose to a peak of 386 +/- 44 pg/ml at the nadir of blood pressure (mean arterial pressure = 47.5 mmHg). ACTH showed a similar pattern, increasing from 49 +/- 10 to a peak of 518 +/- 56 pg/ml. Cortisol values reached their peak of 18.2 +/- 2.5 micrograms % during the recovery phase. beta-Endorphin values displayed a close inverse correlation to blood pressure during hemorrhage, but returned to basal levels more rapidly than blood pressure during the recovery period. Plasma ACTH levels rose significantly more slowly than beta-endorphin as the hemorrhage progressed. An equimolar ratio of ACTH and beta-endorphin returned only as levels declined following the hemorrhagic insult. In awake pigs therefore an arterial hemorrhage is accompanied by endorphin release proportional to the decrement in blood pressure, a somewhat retarded buildup of ACTH, and a still later cortisol peak during recovery. PMID- 3034084 TI - The purely epithelioid peripheral nerve sheath tumor. PMID- 3034085 TI - Purification of Cryptosporidium oocysts and sporozoites by cesium chloride and Percoll gradients. AB - The lack of an adequate system for the in vitro cultivation of Cryptosporidium spp. has forced researchers to work on infected feces or tissues. Molecular and immunological analyses of Cryptosporidium stages must be preceded by complex preparatory steps involving the concentration, storage, purification, excystation of oocysts, and purification of sporozoites. This paper describes two new procedures for the purification of Cryptosporidium. The first, consisting of pretreatment of oocysts with sodium hypochlorite followed by concentration using a Percoll gradient, is suitable for nucleic acid analyses. The second, a concentration of untreated oocysts using a Cesium chloride gradient, is suitable for biochemical and immunological studies, but requires "fresh" oocysts. PMID- 3034086 TI - Loss of granule myeloperoxidase during in vitro culture of human monocytes correlates with decay in antiprotozoa activity. AB - Human monocytes maintained in culture lose microbicidal activity against intracellular protozoa which has been correlated with attenuation of the respiratory burst. The granule enzyme myeloperoxidase, which can markedly amplify hydrogen peroxide-dependent antimicrobial activity, is also lost in vitro. Adherent monocytes were examined immediately, 3 and 10-14 days following explantation, for the magnitude of the stimulated respiratory burst and for cellular myeloperoxidase. Fresh cells generated 254 +/- 38 nmol O2-/mg protein as compared to a peak of 782 +/- 45 nmol O2-/mg at 3 days and less than 100 nmol O2 /mg after 10-14 days. The myeloperoxidase content of the cells also decreased; over 85% was lost after 3 days. Fresh monocytes killed over 90% of ingested Toxoplasma gondii or Leishmania major. In contrast, 10-14 day explanted monocytes killed only 12% of ingested Toxoplasma and 33% of Leishmania, and surviving organisms replicated readily. The 3-day monocytes were significantly less able to kill protozoa than were fresh cells despite their nearly 3-fold greater generation of O2-. If peroxidase was reintroduced into 3-day monocytes by coating organisms with eosinophil peroxidase prior to phagocytosis, their antiprotozoa activity was nearly restored to that of freshly explanted cells. PMID- 3034087 TI - Natural infection of humans, animals, and phlebotomine sand flies with the Alagoas serotype of vesicular stomatitis virus in Colombia. AB - Five isolations of the Alagoas serotype of vesicular stomatitis virus (Rhabdoviridae: Vesiculovirus) were made from naturally infected phlebotomine sand flies (Lutzomyia spp.) collected in Colombia. These are the first isolations of Alagoas virus from an arthropod. Replication of the virus occurred in laboratory-reared sand flies (Lutzomyia longipalpis) after inoculation. Bite and transovarial transmission of the virus was also demonstrated in experimentally infected sand flies. Alagoas virus neutralizing antibodies were found in sera of humans and animals living near the insect collection site; antibody rates among human residents of two nearby towns were 63% and 83%, respectively. Results of comparative serologic studies demonstrated that Alagoas virus is closely related antigenically to Indiana, Cocal, and Maraba viruses and that these four agents form a complex within the vesicular stomatitis virus serogroup. The antigenic similarity among these four viruses makes their differentiation difficult; it also raises doubts about the accuracy of current laboratory methods used for identifying isolates in this serogroup. A discussion follows on the significance of human antibodies to these agents and on the role of sand flies in their ecology. PMID- 3034088 TI - Viremia and immune response with sequential phlebovirus infections. AB - Four groups of hamsters were infected sequentially with various combinations of Arumowot, Chagres, and Gabek Forest viruses. Following each infection, the survival, level of viremia, and immune response of the animals were monitored. All of the agents produced viremia in the hamsters, regardless of the order of their administration. The antibody response, as measured by plaque reduction neutralization test, was monotypic even after two consecutive phlebovirus infections. Arumowot and Chagres viruses produced nonfatal infections in adult hamsters, which were characterized by viremia of several days duration and subsequent antibody formation. In contrast, Gabek Forest virus produced a fulminating and rapidly fatal disease in phlebovirus nonimmune animals. In hamsters previously infected with Chagres and/or Arumowot viruses, Gabek Forest infection was less severe, indicating some degree of cross-protection. The degree of cross-protection was in part related to the sequence of previous phlebovirus infections. No evidence of immune enhancement or other immunopathologic events were observed in the animals. PMID- 3034089 TI - Hepatic resection for primary and secondary neoplasms of the liver. AB - Fifty consecutive major hepatic resections were performed for primary and secondary malignant neoplasms of the liver. There were 7 children and 9 adults with primary neoplasms and 34 patients with secondary neoplasms. The mortality rate was 0 percent and the morbidity rate, 14 percent. Postoperative morbidity correlated with operative blood loss. The 5 year survival rates for children and adults with primary neoplasms were 42 percent and 22 percent, respectively, and the 5 year survival rate for adults with secondary neoplasms was 15 percent. Factors such as disease-free interval, number of metastases, and stage of metastases did not influence the postoperative survival rate. Also, there was no difference in survival rate between patients whose metastases were resected by lobectomy or segmentectomy and those whose metastases were resected by wedge resection. PMID- 3034090 TI - [Corticotropin and glucocorticoids of the feto-placental complex in late toxemia of pregnancy]. PMID- 3034091 TI - Assessment of retinoid-induced differentiation of F9 embryonal carcinoma cells with an enzyme-linked immunoadsorbent assay for laminin: statistical comparison of dose-response curves. AB - A convenient procedure, using enzyme-linked immunoadsorbent assay of laminin, to measure retinoid-induced F9-cell differentiation into parietal endoderm was developed. Dose-response curves were fitted with the Allfit program, a statistical method for the analysis and simultaneous comparison of sigmoidal curves, which has been modified for use with a microcomputer. The procedure was standardized with respect to time of retinoid incubation, time-course of laminin production, effects of dibutyryl cAMP, and nature of individual dose-response curves. Retinoic acid produced a half-maximal response at 1.3 nM. Retinol was 175 fold less potent than retinoic acid and required 72 h to effect a maximum response, in contrast to 48 h for retinoic acid. Six oxidized and/or isomerized metabolites of retinoic acid, including 13-cis-retinoic acid, were less potent than retinoic acid, but were more potent than retinol. The dose-response curves had identical slopes with the exception of those obtained with 13-cis-4-oxo- and 4-oxo-16-hydroxyretinoic acids, the only metabolites tested with two structural alterations relative to retinoic acid. Multiple functional group alterations were synergistic in deactivating retinoic acid. The synthetic retinoids 13-cis-N ethylretinamide and 4-hydroxyphenylretinamide and the steroid hormone 1,25 dihydroxycholecalciferol were inactive. PMID- 3034092 TI - Desalting and concentration of proteins in dilute solution using reversed-phase high-performance liquid chromatography. AB - A rapid method for desalting and concentrating dilute protein solutions using short reversed-phase columns (3-4 cm) has been described. The recovery of proteins is usually 90-100%. The method is simple and rapid and allows the desalting and concentration of protein samples simultaneously. A wide variety of proteins in the range up to 80 kDa can be desalted in microgram to milligram amounts, and volumes up to 1 liter can be concentrated to a few milliliters by a single injection. PMID- 3034093 TI - Immobilization of proteins via arginine residues. AB - A new method for activating polyacrylamide beads to bind proteins via arginine residues is described. The linking reagent, 4-(oxyacetyl)phenoxyacetic acid (OAPA), has been synthesized and characterized. OAPA reacts with arginine or N alpha-acetyl-L-arginine with a stoichiometry of 2 to 1. As expected for an arginine-specific reagent, OAPA inactivates horse liver alcohol dehydrogenase in a time-dependent manner, with the rate of this inactivation decreasing sixfold in the presence of 1 mM NADH. The presence of the carboxyl group in the linking reagent allows efficient coupling to aminated polyacrylamide beads. These derivatized beads are capable of binding various proteins via arginine residues in a time- and pH-dependent manner. Capacities range from less than 0.5 mg/ml to greater than 11 mg/ml, depending on the protein. The proteins are bound in a stable linkage, and preblocking the beads with either arginine or N alpha-acetyl L-arginine eliminates all protein binding. Preblocking of the protein ubiquitin with OAPA reduces binding to a level compatible with the amount of underivatized ubiquitin remaining. The specificity, water solubility, negative charge, and linking ability of OAPA make it an especially valuable tool, both as a protein modification reagent and as a linking reagent in preparing specialized affinity chromatographic media. PMID- 3034094 TI - Generation of a murine monoclonal antibody that detects the fos oncogene product. AB - A hybridoma producing a monoclonal antibody (MoAB) recognizing both the cellular and viral forms of fos has been generated by somatic cell hybridization techniques from spleen cells of mice immunized with a synthetic peptide corresponding to amino acids 128-152, a consensus region, of both the v-fos and c fos oncogene products. Three proteins with molecular weights of 55,000, 44,000, and 42,000 were detected by immunoblotting. While MoAB 2G9C3 failed to immunoprecipitate fos from Finkel-Biskis-Jenkins murine osteosarcoma-virus infected fibroblasts, both the 55,000 v-fos protein and the 39,000 cellular protein were coprecipitated using polyvalent rabbit antibodies to the same peptide. Whereas no cell surface membrane expression of fos was detected, after membrane permeabilization by a brief exposure to lysolecithin it was possible to specifically detect internal fos by immunofluorescence flow cytometry. Immunohistochemical staining of FBJ virus-infected cells revealed intense, nuclear staining. PMID- 3034095 TI - Enzyme-based detection of glycoproteins on blot transfers using avidin-biotin technology. AB - Avidin-biotin technology has been employed for the improved nonradioactive detection of glycoproteins on blots. Periodate oxidation of samples on blots converts the glycoprotein-based carbohydrate residues to the corresponding aldehydes. The latter undergo interaction with preformed complexes consisting of either avidin hydrazide or streptavidin hydrazide combined with biotinylated alkaline phosphatase. The sensitivity of the new assay exceeds the previously described enzyme hydrazide method by a factor of at least 10. The approach can be rendered selective for sialoglycoproteins, and approximately 12 sugar-containing bands could be observed in erythrocyte membrane preparations. Problems of nonspecific binding and high levels of background label were alleviated using a nonglycosylated basic protein (lysozyme) for quenching. PMID- 3034096 TI - Separation of blood group A-active oligosaccharides by high-pressure liquid affinity chromatography using a monoclonal antibody bound to concanavalin A silica. AB - A column for high-pressure liquid affinity chromatography is prepared by binding a murine monoclonal anti-blood group A antibody of IgM isotype to concanavalin A coated silica particles. The column specifically retards blood group A-active oligosaccharides with the nonreducing immunodominant trisaccharide sequence, GalNAc alpha 1-3(Fuc alpha 1-2)Gal beta 1- ..., and separates three A-active oligosaccharides with different core structures. Retention of the oligosaccharides on the column diminishes with increasing temperatures, permitting thermal elution in the range 25-50 degrees C. PMID- 3034097 TI - Comparison of sample preparation methods for the high-performance liquid chromatographic analysis of cell culture extracts for triphosphate ribonucleosides and deoxyribonucleosides. AB - We compared four different procedures for the purification and concentration of nucleoside triphosphates in cell extracts prior to HPLC analysis. Two methods involved precipitation, with either acetonitrile or calcium fluoride. The acetonitrile procedure yielded reasonable recovery and sufficient purity for the subsequent HPLC analysis. The calcium fluoride coprecipitation procedure gave both good recovery and purity; but the recovery was shown to be dependent on the concentration of the nucleoside triphosphates. The other two methods involved small Sep-Pak cartridges. The silica cartridge procedure yielded unfavorable recoveries in periodate-treated cell extracts, apparently due to poor solubility of nucleoside triphosphates in the requisite solvents. The strong anion exchange cartridge procedure yielded both good recovery and purity. This procedure was found to be fast, efficient, and reliable for purifying and concentrating nucleotides in cell extracts. PMID- 3034098 TI - Direct mapping of rare messenger RNAs by means of oligomer-directed ribonuclease H cleavage. AB - A method has been developed for characterizing rare messenger RNAs in the bulk population by using oligodeoxyribonucleotide: RNA hybrids as substrates for Escherichia coli ribonuclease H. Two 1.3-kb mRNAs in lymphocyte cytoplasm, interferon-gamma (0.002% of polyadenylated mRNA), and prothymosin-alpha, have been studied. Interferon-gamma mRNA was cut virtually completely into two fragments, each about 0.6 kb in length, by using an interferon-specific 24-mer to direct cleavage. Prothymosin-alpha mRNA in the same bulk population was unaffected by this treatment. When the 24-mer was replaced by a 12-mer, whose sequence was based on an incomplete cDNA clone for prothymosin-alpha, the products included two fragments of prothymosin-alpha mRNA. The sum of the fragment lengths equaled the length of the mRNA. Although the reaction directed by the smaller oligomer did not go to completion, the 12-mer, and hence the cDNA clone from which it was derived, could nevertheless be oriented with respect to prothymosin-alpha mRNA. With this technique, sequences in mRNA can be mapped without first isolating full-length cDNA clones. PMID- 3034099 TI - Plasma membrane-associated phosphatase activities hydrolyzing [32P]phosphotyrosyl histones and [32P]phosphatidylinositol phosphate. AB - We describe a procedure of preparing [32P]phosphotyrosyl histones with minimal contamination by 32P-labeled lipids; the latter was usually found to be mixed with the phosphoproteins when the cell membrane-enriched fraction of A-431 cells was used as a source of tyrosine kinase. The phosphatase activities previously found to be associated with the plasma membranes of a human astrocytoma were resolved using purified [32P]phosphotyrosyl histones and [32P]phosphatidylinositol phosphate. In comparison with the phosphotyrosyl protein phosphatase, the phosphatidylinositol phosphate phosphatase activity is more active over a broad range of pH values, and its activity is inhibited by fluoride, zinc chloride, and lower concentrations of vanadate. PMID- 3034100 TI - Fluorescent inhibitor probes of enzyme active site conformation: anion binding to angiotensin-converting enzyme. AB - Dansylated tight-binding inhibitors are effective fluorophoric probes for detecting conformational changes of enzyme active sites. In this study they have been employed to examine the effect of anions on the conformation of angiotensin converting enzyme. The efficiency of radiationless energy transfer between enzyme tryptophan residues and an active site-bound dansyl inhibitor has been shown to be enhanced by the addition of chloride. Half-maximal fluorescence enhancement occurs at about 2 mM chloride and is the same for both N-(1-carboxyl-5 dansylamino-pentyl)-glycyl-L-phenylalanine [Ki,app = 50 nM (pH 7.5, 300 mM NaCl)] and N-(1-carboxyl-5-dansylamino-pentyl)-glycyl-L-lysine (Ki,app = 5.7 nM). Other activating anions also evoke similar increases in enzyme-inhibitor energy transfer. Fluorescence changes are not due to binding additional inhibitor molecules but rather to an anion-induced change in protein conformation. PMID- 3034101 TI - Identification and purification of a cytosolic phosphotyrosyl protein phosphatase from bovine spleen. AB - A protein tyrosine kinase with an apparent Mr of 60,000 was highly purified from bovine spleen and used to phosphorylate poly(Glu, Tyr) (4:1) on tyrosine residues for the study of phosphotyrosyl protein phosphatases from this tissue. About 70% of the phosphotyrosyl protein phosphatase activity in extracts of bovine spleen was adsorbed on DEAE-Sepharose. Chromatography of the eluted phosphotyrosyl protein phosphatases on phosphocellulose indicated the presence of at least two species, one that did not bind to the phosphocellulose and a second species that did bind and was eluted at about 0.5 M NaCl. The phosphatase that did not bind to phosphocellulose was further purified by successive chromatography on poly(L lysine)-Sepharose, L-tyrosine-agarose, poly(Glu,Tyr)-Sepharose, and Sephacryl S 200. The enzyme had an apparent Mr of 50,000 as estimated by gel filtration and 52,000 as estimated by NaDodSO4- polyacrylamide gel electrophoresis. The phosphatase exhibited a pH optimum of 6.5-7.0, was inhibited by Zn2+ and vanadate ions, and was stimulated by EDTA. Sodium fluoride and sodium pyrophosphate, inhibitors of phosphoseryl protein phosphatases, had no effect on the enzyme. Protein inhibitors of type 1 phosphoseryl/threonyl phosphatase were also ineffective. PMID- 3034102 TI - Effect of structural modification at carbon atom 1 of leukotriene B4 on the chemotactic and metabolic response of human neutrophils. AB - Human neutrophils biosynthesize the chemoattractant leukotriene B4 (LTB4) and metabolize LTB4 to omega oxidative products 20-hydroxy-LTB4 (20-OH-LTB4) and 20 carboxy-LTB4 (20-COOH-LTB4). In this study, we prepared the C-1 methyl ester and N-methyl amide of LTB4 and then examined neutrophil chemotaxis and metabolism of these derivatives of LTB4. The results show that chemical modification of LTB4 at carbon atom 1 dramatically affects metabolism of the lipid molecule. The free acid form of LTB4 was taken up and metabolized by human neutrophils, while the methyl ester and N-methyl amide derivatives were poor substrates for omega oxidation. Although human neutrophils were poorly attracted to the methyl ester of LTB4, the amide derivative was a complete agonist of the neutrophil chemotactic response and displayed an ED50 for chemotaxis identical to that of LTB4. Therefore, we concluded that omega oxidation is not a requirement for the neutrophil chemotactic response induced by LTB4. These results also indicate that the N-methyl amide of LTB4 may be a useful ligand for the elucidation of molecular mechanisms operative in neutrophil chemotaxis to LTB4, since the C-1 derivative is not further metabolized. Two separate responses of human neutrophils are elicited by LTB4, resulting in both cellular activation and generation of omega oxidation products. It appears that putative receptors on the neutrophils can distinguish between LTB4 and certain derivatives that are structurally identical except for modification at the C-1 position (i.e., the methyl ester). LTB4 derivatives modified at the C-1 position do not undergo conversion to omega oxidation products by the neutrophil. PMID- 3034103 TI - Assaying for superoxide dismutase activity: some large consequences of minor changes in conditions. AB - Most assays for superoxide dismutase depend upon competition between the enzyme and some indicating scavenger for O-2. We have investigated the effects of experimental variables on assays based upon the use of either ferricytochrome c or nitro blue tetrazolium. Our results should help investigators to avoid the numerous potential pitfalls which necessarily surround these assay methods. PMID- 3034104 TI - Maturation antigen of the mouse sperm flagellum: II. Origin from holocrine cells of the distal caput epididymis. AB - During epididymal transit, the mouse sperm flagellum acquires a surface glycoprotein (SMA4) from epididymal fluid that functions as a sperm antiagglutinin. To determine the origin of this molecule, testes and epididymides of male mice were sectioned for light microscopy and stained with wheat germ agglutinin (WGA)-peroxidase, a probe that has been used previously to examine the biology of SMA4. WGA reactivity was localized to the cytoplasm in a small population of cells in the distal caput epididymis. Testis cells and principle cells of the caput were nonreactive with WGA, while stereocilia were stained on principle cells in the corpus and cauda. The WGA-positive cells in the distal caput were identified as holocrine cells on the basis of morphology, distribution, and PAS + reaction. At high magnification, intense WGA reactivity was due to the presence of numerous apical granules in the cytoplasm. The location of the cells in distal caput coincided exactly with the region of tubule in which sperm first acquired SMA4 on their flagellae. These data suggest that holocrine cells near the junction of caput and corpus epididymis are the source of the sperm antiagglutinin SMA4. PMID- 3034105 TI - The pattern of train-of-four fade after atracurium: influence of different priming doses. AB - This study was designed to investigate the effect of three different priming doses of atracurium--0.06, 0.07, and 0.08 mg/kg--followed 3 min later by the remainder of a 0.5 mg/kg dose on the relationship between the depression in the first twitch of the train-of-four (T1) and train-of-four (TOF) fade. This relationship was studied after the administration of the full dose of the relaxant in all groups. Of all the priming doses, 0.08 mg/kg atracurium, when followed 3 min later by 0.42 mg/kg atracurium, had a significantly greater fade in the TOF ratio at any given T1 value. This may indicate significant prejunctional activity. Acceleration of the onset of neuromuscular blockade was, however, evident in all groups that received atracurium in divided doses. The implication is, therefore, that prejunctional activity may not contribute significantly to the acceleration of onset of neuromuscular blockade after administration of atracurium in divided doses, as described in this study. PMID- 3034106 TI - Sodium bicarbonate attenuates pain on skin infiltration with lidocaine, with or without epinephrine. PMID- 3034107 TI - Endocrinological changes following etomidate, midazolam, or methohexital for minor surgery. AB - Etomidate is known to inhibit adrenocorticosteroid synthesis. The extent and duration of the effects of etomidate (63 +/- 6.4 mg) on spontaneous and stimulated corticosteroid levels, as well as on plasma concentrations of ACTH, beta-endorphin, and catecholamines were examined and compared to those following administration of the new benzodiazepine, midazolam, or of methohexital. Twenty nine healthy, young, male orthopedic patients were randomized into three groups receiving either etomidate/fentanyl (n = 12), midazolam/fentanyl (n = 8), or methohexital/fentanyl (n = 9). Etomidate caused cortisol levels to decrease from 12.5 +/- 1.2 micrograms/dl preoperatively to 5.9 +/- 0.8 micrograms/dl after operation (P less than 0.001), compared to an increase from 12.0 +/- 1.9 micrograms/dl to 18.5 +/- 2.9 micrograms/dl in the group receiving methohexital. At 6 and 20 h postoperatively, all cortisol levels were normal. The cortisol decrease from 12.5 +/- 1.7 to 7.6 +/- 1.5 caused by midazolam was similar to that following etomidate, but the response to exogenous ACTH was significantly impaired in patients receiving etomidate as compared to those receiving midazolam. ACTH and beta-endorphin levels increased in patients receiving etomidate, presumably as a result of the interruption of negative feedback due to cortisol synthesis inhibition. Midazolam on the other hand prevented the increase of ACTH and beta-endorphin levels. Etomidate completely suppressed spontaneous aldosterone levels (from 33 +/- 6.7 to 7 +/- 2.1 pg/ml), as well as the response to stimulation with exogenous ACTH without affecting serum electrolytes. Etomidate had no influence on plasma catecholamines, but midazolam attenuated the stress-related epinephrine increase. PMID- 3034108 TI - Inhibition by methylprednisolone of zymosan-induced leukotriene synthesis in alveolar macrophages. AB - Alveolar-macrophage-derived 5-lipoxygenase metabolites of arachidonic acid (AA) are thought to be important mediators of lung inflammation and injury. Inhibition of AA metabolism may be an important anti-inflammatory mechanism of glucocorticoid (GC) action. In the present study, we have examined the effect of methylprednisolone (MP) on leukotriene (LT) B4 and LTC4 synthesis by cultured rat alveolar macrophages (AMs) stimulated with the proinflammatory particle zymosan. Zymosan stimulation resulted in the formation of LTB4 greater than thromboxane B2 (TxB2) greater than LTC4 as assessed by both high performance liquid chromatography and radioimmunoassay. Sixteen hours of pretreatment with 1 microM MP maximally inhibited synthesis of TxB2 by 76 +/- 4%, LTB4 by 83 +/- 3%, and LTC4 by 91 +/- 3%. The inhibition of eicosanoids was specific, as the sex hormones progesterone, testosterone, and beta-estradiol had no consistent effect. Inhibition was dose-dependent, with 50% inhibition of synthesis occurring at 10( 7) M for TxB2, but at 10(-8) M for both LTB4 and LTC4. Pretreatment with MP for only 1 h inhibited LTC4 production (42 +/- 3%) to a greater extent than either TxB2 (25 +/- 1%) or LTB4 (14 +/- 7%). These data indicate that MP significantly and substantially inhibits zymosan-induced LTB4 and LTC4 synthesis. Furthermore, the greater degree, greater potency, and faster onset of action for MP inhibition of LT synthesis than of TxB2 synthesis suggest the possibility that MP may exert postphospholipase effect(s) on LT synthesis. These results support the possibility that modulation of AM arachidonate metabolism may represent an important anti-inflammatory mechanism of GC action in the lung. PMID- 3034109 TI - Regulation by the antioxidants ascorbate, cysteine, and dapsone of the increased extracellular and intracellular generation of reactive oxidants by activated phagocytes from cigarette smokers. AB - The bimodal pattern of N-formyl-methionyl-leucyl-phenylalanine (FMLP)-activated luminol-enhanced chemiluminescence with distinct early (occurring within 1 min) extracellular and late intracellular oxidative responses was compared in polymorphonuclear leukocytes (PMNL) from asymptomatic cigarette smokers and nonsmoking control subjects. Relative to control PMNL, the PMNL from smokers were hyperreactive to FMLP stimulation with increased generation of both extracellular (p less than 0.025) and intracellular (p less than 0.025) reactive oxidants. Smokers' PMNL also showed increased PMNL-activated superoxide generation and increased apparent receptors for FMLP. The water-soluble antioxidants ascorbate and cysteine (2.5 X 10(-5) M to 2.5 X 10(-4) M) selectively neutralized the extracellular activity of PMNL-derived reactive oxidants. The lipid-soluble antioxidant dapsone (1.25 to 30 micrograms/ml), on the other hand, inhibited both the extracellular and intracellular FMLP-activated chemiluminescence responses in PMNL from smokers and nonsmoking control subjects. Regulation of the increased extracellular and intracellular membrane-associated oxidative responses in PMNL from cigarette smokers is probably an important function of water-soluble and lipid-soluble antioxidants in vivo. PMID- 3034111 TI - Bone marrow examination in small cell lung cancer. PMID- 3034110 TI - Liquoid in blood cultures. PMID- 3034112 TI - [Oncogenes]. PMID- 3034113 TI - [Neuropathy caused by cisplatin. 7 cases including one with an autopsy study]. AB - The clinical, electrophysiological and histopathological features in seven cases of cisplatinum peripheral neuropathy are reported and compared with the literature data. The neuropathy appears for an average intake of 500 mg/m2 of DDP. The symptoms are those of a symmetric, distal, predominantly sensitive neuropathy of an axonal type with major involvement of proprioception. Neurological improvement is poor after withdrawal of the drug. A post mortem study performed in one case showed a degeneration of the posterior column in the cord and residual nodules of Nageotte in a lumbar spinal ganglion. The systematic study of the tendon reflexes and distal pallesthesia in subjects treated with the drug, may reveal the neuropathy before the onset of the most disabling symptoms (paresthesia, ataxia, pain, Lhermitte's sign). PMID- 3034114 TI - Elevation in hypothalamic cyclic AMP as a common factor in the facilitation of lordosis in rodents: a working hypothesis. PMID- 3034115 TI - Cytochrome oxidase deficiency: clinical and biochemical heterogeneity. PMID- 3034116 TI - Ca2+ signalling in exocrine glands in comparison to that in vascular smooth muscle cells. PMID- 3034117 TI - Mitochondrial myopathies involving the respiratory chain: a biochemical analysis. PMID- 3034118 TI - Altered sorbitol and myo-inositol metabolism as the basis for defective protein kinase C and (Na,K)-ATPase regulation in diabetic neuropathy. PMID- 3034119 TI - Mechanisms whereby insulin and other hormones binding to cell surface receptors influence metabolic pathways within the inner membrane of mitochondria. PMID- 3034120 TI - Characterization of mediators of insulin action. PMID- 3034121 TI - Mechanism of receptor kinase action on membrane protein recycling. PMID- 3034122 TI - The ADP/ATP carrier as a mitochondrial auto-antigen--facts and perspectives. PMID- 3034123 TI - Transmembrane signalling by growth factors. PMID- 3034124 TI - [Cronkhite-Canada syndrome]. PMID- 3034125 TI - [Glucagonoma syndrome. Unusual histologic image]. PMID- 3034126 TI - [Multiple trichoepitheliomas, cylindromas and milia. An entity]. AB - Four patients from two different families presented with multiple papular trichoepitheliomas of the face associated with cylindromas of the scalp and, in one of them, milium. This association, first described by Adamson, has now become classical. It is transmitted as an autosomal dominant trait with variable penetrance. Histochemical studies gave the following results: ATPase negative in the two types of tumour, phosphorylase weakly positive, NADH diaphorase positive in the basal cells of the trichoepitheliomas and diffusely in cylindromas. These results suggest that the cylindromas are of apocrine origin. Using monoclonal antibodies, it has been possible to demonstrate the presence of Langerhans cells in both trichoepitheliomas and cylindromas. The BL9 and KL3 antikeratinocyte monoclonal antibodies were negative, whereas the KL3 antibody, which recognizes the 55-57 Kd polypeptides of keratin, marked the suprabasal part of the tumours. These results are in favour of incomplete cell differentiation. Treatment with retinoids was ineffective, as in all other cases reported. Electrocoagulation or surgical excision om request for cosmetic reasons seem to be only possible treatments. PMID- 3034127 TI - [OKT6-labelled non-X histiocytosis (generalized eruptive histiocytoma?) with intermediate vimentin filaments. Apropos of a case]. PMID- 3034128 TI - [Human Epstein-Barr virus infection. Dermatological manifestations]. PMID- 3034129 TI - [Apropos of rheumatoid purpura and human parvovirus]. PMID- 3034130 TI - Effect of a preliminary infection of ducks with EDS-76 virus on the Derzsy's disease pathology. AB - The aim of this work was to investigate the role of a non pathogenic adenovirus (EDS-76, strain A-127), frequently isolated from ducklings on the infection by a pathogen agent (such as the parvovirus responsible for Derzsy's disease, DDV). Three groups of ducklings were respectively infected with A-127 alone, at day old, DDV alone at day 10th, A-127 at day old plus DDV at day 10th. The infected groups and a control one were raised under similar conditions. Pathological signs were observed and body weights recorded. Sera were collected weekly and analysed for DDV and A-127 antibodies. In addition, in vitro assays of the multiplication of these viruses and studies on cytopathic effects on duck embryo fibroblasts were conducted. The humoral response did not appear to be influenced by dual infection. No differences in cytopathogenic effects were detected in vitro between single and dual infection. On the other hand, in ducklings, dual infection increased the mortality rate and depressed the body weight gain, as compared to infection with either virus alone. The drastic effect on the body weight gain of the EDS-76 virus in the presence of DDV clearly shows a synergic role of this virus on the course of Derzsy's disease. PMID- 3034131 TI - [Demonstration of BVD virus (bovine diarrhea virus) in many cell lines]. AB - By specific fluorescent antibodies, it was shown that cell lines from bovine origin were contaminated with BVD virus. The same virus was also detected in cell lines from different species like horse, cat, monkey, rabbit, pig and sheep. The authors express their concern about the use of these cell lines as substrate for live virus vaccines. PMID- 3034132 TI - Liposarcoma of pleural cavity with recurrence as malignant fibrous histiocytoma. AB - The case of a 54-year-old male, with a massive right pleural liposarcoma weighing over 3200 g, is presented. The tumor was found by light and electron microscopy to be of well-differentiated and pleomorphic subtypes, and it apparently represents the sixth reported case of liposarcoma primary to the pleura. Two years after excision of the primary tumor, it recurred as a neoplasm with histologic and ultrastructural features characteristic of malignant fibrous histiocytoma. The histogenetic and pathologic implications of the above findings are discussed, and the literature regarding intrathoracic liposarcoma and malignant fibrous histiocytoma is reviewed. PMID- 3034134 TI - Chronic Epstein-Barr virus infection. AB - The Epstein-Barr virus (EBV) has been associated with classic infectious mononucleosis, Burkitt's lymphoma, nasopharyngeal carcinoma, and B-cell lymphomas in primary and secondary immunodeficiency disease. The availability of specific serologic diagnosis of EBV, rather than dependence on heterophile antibody positivity, has broadened the scope of EBV-associated diseases. A chronic neuroasthenia syndrome accompanied by antibody titers to the viral capsid antigen and early antigen of EBV, which are higher than found in asymptomatic individuals, is one such additional EBV-associated syndrome. This paper describes the clinical and laboratory responses to EBV that are present in this chronic syndrome. It then discusses management of these patients and the difficulties in establishing a cause-and-effect relationship between EBV and chronic neuroasthenia along with recommendations for future studies. PMID- 3034135 TI - Rotavirus gastroenteritis. AB - Rotaviruses have proven to be major causes of pediatric diarrheal disease morbidity and mortality worldwide. The past several years have yielded substantial new insights into these viruses, their epidemiology, and the mechanisms of host resistance to them. These insights have in turn resulted in development of several candidate rotavirus vaccine strains, bringing closer the possibility of effective protection of young children from rotavirus disease. PMID- 3034133 TI - [Evaluation of a new therapy (beidellian montmorillonite--guar gum) in the treatment of constipation]. PMID- 3034136 TI - Live attenuated varicella vaccine. AB - Live attenuated varicella vaccine will in all probability soon be licensed in the United States for immunization of healthy children and adults and certain immunocompromised children. This vaccine can be expected to protect susceptibles from varicella (or to modify it) in those subsequently exposed to the wild virus. The vaccine is safe, immunogenic, and is not associated with an increase in the incidence of zoster. PMID- 3034137 TI - Endogenous digitalis-like natriuretic factors. AB - There is increasing evidence for endogenous, circulating compounds that interact with the digitalis receptor of [Na,K]ATPase and with antidigoxin antisera. Circulating levels of these digitalis-like compounds increase in response to fluid or salt loading and appear to play a role in diseases characterized by fluid and salt retention, e.g. renal failure, liver disease, acromegaly, experimental and human hypertension, and preeclampsia. Because of assay nonspecificity, many diverse substances are being measured. Of the few compounds currently identified as having "digitalis-like" activity, none appears to be the natural ligand of the digitalis receptor and none appears linked with hypertension. Nevertheless, research still suggests that digitalis-like factors may have a central role in essential hypertension and related disorders. PMID- 3034138 TI - Metabolism of alpha- and beta-adrenergic receptors in vitro and in vivo. AB - Despite considerable evidence that changes in number of adrenergic receptors can occur under various conditions, knowledge of the mechanisms mediating these changes is still rudimentary. As discussed, indirect approaches emphasizing the kinetics of receptor turnover have been the principal means of investigation. These indirect methods, which depend on the ability of a radioligand to detect the receptors, are limited by several factors. Even so, the data obtained using indirect approaches, in particular on various model systems in cell culture, lead to several conclusions: Both alpha 1- and beta-adrenergic receptors are metabolized rather slowly in vitro under basal conditions, in the absence of exposure to agonists. Typical half-lives are greater than 20 hr, a turnover that is slower than that of several other classes of neurotransmitter and hormone receptors (9, 10, 12, 16). Moreover, alpha 1-adrenergic receptors and beta adrenergic receptors can have substantially different half-lives, even when expressed on the same cell. In view of the relatively slow rate of disappearance of adrenergic receptors under basal conditions, settings in which receptor number increases are almost certainly to result from increases in one or more of the factors that contribute to the rate of receptor appearance on the plasma membrane. Treatment of cells with agonists markedly shortens the half-life of alpha 1- and beta-adrenergic receptors. This shortened half-life results primarily from an enhanced loss of receptors from the plasma membrane, and not from agonist-induced attenuation of receptor appearance. In fact, data acquired from studies of receptor recovery after agonist-induced down-regulation suggest that rates of receptor reappearance are markedly enhanced through either receptor recycling or an increase in receptor synthesis. Limited studies conducted in vivo yield qualitatively similar results to those observed in in vitro studies of the metabolism of adrenergic receptors. In general, adrenergic receptors in the CNS turn over more slowly than those in peripheral tissues. These conclusions help to highlight the many aspects of metabolism of adrenergic receptors that are as yet unknown, including identification and characterization of the cellular machinery responsible for receptor metabolism, elucidation of the molecular events that control metabolism, and assessment of how drugs and other factors influence these events. Future studies are likely to be based on the development of new methodology with antireceptor antibodies, receptor cDNA's, and improved morphological methods (autoradiography, immunohistochemistry, etc).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3034139 TI - Prostaglandins, leukotrienes, and platelet-activating factor in shock. AB - Three major lines of evidence support a role of eicosanoids and PAF in shock. Formation of each of the cyclooxygenase metabolites of arachidonate is enhanced at some point during the shock; these metabolites include PGE2, PGF2 alpha, PGI2, and TXA2. Enhanced formation of 5-HETE and the cysteinyl-LTs provides evidence for activation of the 5-lipoxygenase pathway of arachidonate metabolism, and preliminary biochemical evidence suggests that formation of PAF in anaphylactic and endotoxic shock is also enhanced. Second, TXA2, cysteinyl-leukotrienes, and, to an even greater extent, PAF are able to produce shock and death in intact animals. Third, pharmacological studies show that selective antagonists or synthesis inhibitors modify the course of the shock. While any of these lines of evidence may not by itself provide proof for a cause-effect relationship, the data taken together strongly suggest that vasoactive lipids might be involved in fundamental processes in the pathophysiology of shock. However, the role of vasoactive lipids might vary in different shock paradigms, change at various time points during the evolution of the shock, and depend on the species studied. Moreover, while the majority of the reports tend to focus on a specific substance, the metabolism of all of the eicosanoids mentioned, as well as PAF and probably other arachidonate metabolites (e.g. 15-lipoxygenase products such as lipoxins), changes during shock states. This fact probably causes most of the discrepancies in studies using specific antagonists or synthesis inhibitors to modify the state of shock. Thus, while blockade of one mediator might provide some protection, it might not be sufficient to halt or reverse the main course of the pathophysiological process. For example, the increase in vascular permeability, a fundamental phenomenon in trauma, anaphylaxis, or endotoxemia, might be mediated by PAF, LTs, PGs, peptides (e.g. kinins, substance P, CGRP) and amines (e.g. histamine in some species). Attempting to reverse such a complex phenomenon by blocking one specific factor might not be productive unless the specific substance played a key role in generation of the other factors. It seems, however, that while interactions between PGs, LTs, and PAF do occur (31, 32, 70), none of the shock states are crucially dependent on one class of the vasoactive lipids. Therefore, the therapeutic strategy should be based on multiple sites of action, either by drug combinations or multiple actions of a specific drug.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3034141 TI - Biochemical and molecular genetic analysis of hormone-sensitive adenylyl cyclase. PMID- 3034140 TI - Calcium channel ligands. PMID- 3034142 TI - Human health effects of polychlorinated biphenyls (PCBs) and polybrominated biphenyls (PBBs). PMID- 3034145 TI - In vitro activity of a new cyclic lipopeptide antibiotic, LY146032, against gram positive clinical bacteria. AB - The in vitro activity of LY146032, a novel cyclic lipopeptide antibiotic, was tested against different gram-positive clinical isolates. The activity of LY146032 was clearly higher than that of vancomycin against all isolates tested. However, in some instances rifampin and imipenem showed higher activity than did LY146032. PMID- 3034143 TI - Demonstration of viral thymidine kinase inhibitor and its effect on deoxynucleotide metabolism in cells infected with herpes simplex virus. AB - The thymidine analog 5'-ethynylthymidine was a potent inhibitor of herpes simplex virus type 1 (strain KOS)-induced thymidine kinase with a Ki value of 0.09 microM. 5'-Ethynylthymidine was less inhibitory against herpes simplex virus type 2 (strain 333)-induced thymidine kinase with a Ki of 0.38 microM and showed no inhibition against human cytosolic thymidine kinase under the conditions tested. The compound was effective against the altered thymidine kinase induced by acyclovir- and bromovinyldeoxyuridine-resistant virus variants. At 100 microM 5' ethynylthymidine, the cellular pool size of dTTP in herpes simplex virus type 1 infected cells was 5% that of infected cells receiving no drug treatment, while there was no significant effect on the pool sizes of dATP, dGTP, and dCTP. There was a positive correlation between dTTP pools and the intracellular thymidine kinase activity of herpes simplex virus type 1-infected cells. When tested alone, 5'-ethynylthymidine exhibited no antiviral activity, but it antagonized the antiviral efficacy of five compounds which require viral thymidine kinase for their action. PMID- 3034144 TI - Identification of porins in outer membrane of Proteus, Morganella, and Providencia spp. and their role in outer membrane permeation of beta-lactams. AB - Proteus mirabilis, Proteus vulgaris, Morganella morganii, Providencia rettgeri, and Providencia alcalifaciens, which were once classified into the same genus, Proteus, were studied. Cefoxitin-resistant mutants from these species were isolated, and it was confirmed that the resistance was attributed to the lack of an outer membrane protein, resulting in a significant decrease in the penetration of hydrophilic cephalosporins through the outer membrane. Comparison of the mutant strains with their parental strains in the diffusion rates of six monoanionic cephalosporins, a zwitterionic cephalosporin (cephaloridine), and a divalent anionic cephalosporin (cephalosporin C) suggested that each species had only one kind of porin protein, with molecular weights of 40,000 (Proteus mirabilis) or 37,000 (the other four species) and that the porins formed channels with cation selectivity, except for Proteus vulgaris. Porin proteins were purified from all the bacterial species except Providencia alcalifaciens, and the radius of the pores formed by the purified porins was estimated by the use of the liposome swelling assay. The pore radii were estimated to be approximately 0.59 nm (Proteus mirabilis), 0.63 nm (Proteus vulgaris), 0.58 nm (Providencia rettgeri), and 0.60 nm (M. morganii), similar to the size of the pore radius of Escherichia coli porins. PMID- 3034146 TI - Effect of calcium on in vitro activity of LY146032 against Clostridium difficile. AB - The in vitro MICs of LY146032 against 63 isolates of Clostridium difficile tested in Wilkins-Chalgren broth ranged from 0.5 to greater than 32 micrograms/ml, with MICs of 4 and 8 micrograms/ml for 50 and 90% of the isolates, respectively. However, when the test medium was supplemented with physiologic concentrations of calcium, the MIC for 90% of the isolates was reduced to less than or equal to 0.12 microgram/ml. PMID- 3034147 TI - Relationship between structure and antiviral activity of 5-methoxymethyl-2' deoxyuridine and 5-methoxymethyl-1-(2'-deoxy-beta-D-lyxofuranosyl)uracil. AB - 5-Methoxymethyl-1-(2'-deoxy-beta-D-lyxofuranosyl)uracil (MMdLU) was not active against the herpes simplex viruses. The relationship between molecular conformation and antiviral activity for the two epimers, 5-methoxymethyl-2' deoxyuridine (MMdUrd) and MMdLU, is discussed. MMdUrd was phosphorylated by the virus-induced deoxythymidine kinase. In contrast, MMdLU did not serve as a substrate for the kinase. The geometry and distance between the 5'-CH2OH and 3' OH groups of the furanose ring appear to be key factors in determining the efficiency of phosphorylation by the virus-induced deoxythymidine kinase, and hence antiviral activity. PMID- 3034148 TI - Antiviral activity of various 1-beta-D-arabinofuranosyl-E-5-halogenovinyluracils and E-5-bromovinyl-2'-deoxyuridine against salmon herpes virus, Oncorhynchus masou virus (OMV). AB - 1-beta-D-Arabinofuranosyl-E-5-bromovinyluracil (BVaraU), 1-beta-D arabinofuranosyl-E-5-iodovinyluracil (IVaraU), 1-beta-D-arabinofuranosyl-E-5 chlorovinyluracil (CVaraU) and 1-beta-D-arabinofuranosyl-5-vinyluracil (VaraU) were examined for antiviral activity against salmon herpesvirus, Oncorhynchus masou virus (OMV) in vitro using Yamame (Oncorhynchus masou) kidney cells (YNK). BVaraU, IVaraU, CVaraU and VaraU were highly active against OMV; 50% inhibitory concentration (IC50): 0.01, 0.003, 0.003, 0.003 microgram/ml, respectively. The IC50 of 5-bromovinyl-2'-deoxyuridine (BVDU) was 0.3 microgram/ml. The lower activity may be due to cleavage of it N-glycosyl linkage by pyrimidine nucleoside phosphorylases (i.e. thymidine phosphorylase) during the incubation period. The arabinofuranosyl counterparts are resistant to this (these) enzyme(s). Both OMV induced DNA polymerase and cellular DNA polymerase alpha were strongly inhibited by BVaraU 5'-triphosphate (BVaraUTP). In an in vivo study, daily immersion of OMV infected chum salmon (Oncorhynchus keta) fry into aqueous solution of BVaraU (5 micrograms/ml, 30 min/day, 30 times) did not increase the life span of infected fish. PMID- 3034149 TI - Antipicornavirus activity of some diaryl methanes and aralkylaminopyridines. AB - Sixteen diarylmethanes and ten aralkylaminopyridines were initially evaluated for their in vitro activity against rhinoviruses 1A, 2 and 64 and against coxsackievirus A21 and for their oral prophylactic and therapeutic activity in mice challenged with coxsackievirus A21. Based on these preliminary studies the diarylmethane (3,4-dichlorophenoxy)-(5 methylsulfonyl-2-pyridinyl)-methane and the aralkylaminopyridine (2-(3,4-dichlorobenzylamino)-5-methylsulfonylpyridine were compared with their oxygen bridged analogue 2-(3,4-dichlorophenoxy)-5 (methylsulfonyl)pyridine for in vitro activity against a larger number of picornaviruses and for their in vivo protective efficacy in dose response assays. All three compounds exhibit similar in vitro activity inhibiting 12 to 15 (52.2 65.3%) of the 23 picornaviruses tested at concentrations of less than 5.0 micrograms/ml. However, the aralkylaminopyridine was found to be the most active in vivo; significantly protecting coxsackievirus A21 challenged mice after a single oral dose of 37.5 mg/kg (P less than or equal to 0.05) and during a continuous oral dose regimen of as low as 18.8 mg/kg per day (P less than 0.01). PMID- 3034150 TI - Antiviral activity of glycyrrhizin against varicella-zoster virus in vitro. AB - One of the plant extracts, glycyrrhizin (GL) was investigated for its antiviral action on varicella-zoster virus (VZV) in vitro. When human embryonic fibroblast (HEF) cells were treated with GL after inoculation of virus (post-treatment), the average 50%-inhibitory dose (ID50) for five VZV strains was 0.71 mM, and the selectivity index (ratio of ID50 for host-cell DNA synthesis to ID50 for VZV replication) was 30. GL was also effective against VZV replication when HEF cells were treated 24 h before the inoculation (pretreatment). Furthermore, at a concentration of 2.4 mM GL inactivated more than 99% of virus particles within 30 min at 37 degrees C. In combination with other anti-herpes drugs (acyclovir, adenine arabinoside, bromovinyldeoxyuridine, and phosphonoformate) or human native beta-interferon, GL had an additive or slightly synergistic effect on VZV replication. The mechanism of anti-VZV action is still unclear. We postulate that GL inhibits the penetration, uncoating or release of virus particles. PMID- 3034152 TI - Transport of lactate and other short-chain monocarboxylates in the yeast Saccharomyces cerevisiae. AB - Saccharomyces cerevisiae IGC4072 grown in lactic acid medium transported lactate by an accumulative electroneutral proton-lactate symport with a proton-lactate stoichiometry of 1:1. The accumulation ratio measured with propionate increased with decreasing pH from ca. 24-fold at pH 6.0 to ca. 1,400-fold at pH 3.0. The symport accepted the following monocarboxylates (Km values at 25 degrees C and pH 5.5): D-lactate (0.13 mM), L-lactate (0.13 mM), pyruvate (0.34 mM), propionate (0.09 mM), and acetate (0.05 mM), whereas apparently a different proton symport accepted formate (0.13 mM). The lactate system was inducible and was subject to glucose repression. Undissociated lactic acid entered the cells by simple diffusion. The permeability of the plasma membrane for undissociated lactic acid increased exponentially with pH, and the diffusion constant increased 40-fold when the pH was increased from 3.0 to 6.0. PMID- 3034151 TI - Molecular cloning of a xylanase gene from Bacteroides succinogenes and its expression in Escherichia coli. AB - A gene coding for xylanase synthesis in Bacteroides succinogenes was isolated by cloning, with Escherichia coli HB101 as the host. After partial digestion of B. succinogenes DNA with Sau3A, fragments were ligated into the BamHI site of pBR322 and transformed into E. coli HB101. Of 14,000 colonies screened, 4 produced clear halos on Remazol brilliant blue-xylan agar. Plasmids from two stable clones recovered exhibited identical restriction enzyme patterns, with the same 9.4 kilobase-pair (kbp) insert. The plasmid was designated pBX1. After subcloning of restriction enzyme fragments, a 3-kbp fragment was found to code for xylanase activity in either orientation when inserted into pUC18 and pUC19. The original clone possessed approximately 10-fold higher xylanase activity than did clones harboring the 3-kbp insert in pUC18, pUC19, or pBR322. The enzyme was partially secreted into the periplasmic space of E. coli. The periplasmic enzyme of the BX1 clone had 2% of the activity on carboxymethyl cellulose and less than 0.2% of the activity on p-nitrophenyl xyloside and a range of other substrates that it exhibited on xylan. The xylanase gene was not subject to catabolite repression by glucose or induction by either xylan or xylose. The xylanase activity migrated as a single broad band on nondenaturing polyacrylamide gels. The Km of the pBX1 encoded enzyme was 0.22% (wt/vol) of xylan, which was similar to that for the xylanase activity in an extracellular enzyme preparation from B. succinogenes. Based on these data it appears that the xylanase gene expressed in E. coli is fully functional and codes for an enzyme with properties similar to the B. succinogenes enzyme(s). PMID- 3034153 TI - Characterization of virucidal agents in activated sludge. AB - A comprehensive study was carried out to determine the properties of agents responsible for loss of virus infectivity in mixed-liquor suspended solids (MLSS) of activated sludge. Initial experiments revealed that model enteric viruses (poliovirus-1 and rotavirus SA-11) were irreversibly inactivated in MLSS and released their RNA genomes. Enteric viruses belonging to other genera (echovirus 12, coxsackievirus A13, reovirus-3) were also shown to lose infectivity in MLSS. Although the virucidal activity decreased at reduced temperatures, MLSS still retained significant activity at 4 degrees C. The virucidal agents in MLSS were stable for months at 4 degrees C, but their activity decreased approximately 50% during 4 days of aeration at 26 degrees C. Primary effluent, the nutrient source for activated sludge, also contained virucidal activity. After centrifugation of MLSS, almost all virucidal activity was found in the particulate fraction because of inhibitory substances retained in the supernatant fraction. Decreasing or increasing the solids concentration of the particulate fraction did not increase the virucidal activity of the fraction. The effects of heat and antibiotics on the virucidal activity of MLSS, coupled with the finding that the activity can be produced in autoclaved primary effluent seeded with MLSS, strongly support the conclusion that microorganisms are responsible for this activity. Attempts to characterize the virucidal microbial components of MLSS indicated that treatments that resulted in the inactivation or removal of microorganisms also caused a loss of virucidal activity. Thus, it appears that the virucidal components of microorganisms are either short-lived or active only while bound to the organisms themselves. PMID- 3034155 TI - Fate of viruses in artificial wetlands. AB - Little is known about the ability of wetlands to remove disease-causing viruses from municipal wastewater. In this study we examined the survival of several indicators of viral pollution (indigenous F-specific bacteriophages, seeded MS2 bacteriophage, and seeded human poliovirus type 1) applied in primary municipal wastewater to artificial wetland ecosystems. Only about 1% of the indigenous F specific RNA bacteriophages survived flow through the vegetated wetland beds at a 5-cm-day-1 hydraulic application rate during the period from June through December 1985. The total number of indigenous F-specific bacteriophages (F specific RNA and F-specific DNA phages) was also reduced by about 99% by wetland treatment, with the mean inflow concentration over the period of an entire year reduced from 3,129 to 33 PFU ml-1 in the outflow of a vegetated bed and to 174 PFU ml-1 in the outflow of an unvegetated bed. Such superior treatment by the vegetated bed demonstrates the significant role of higher aquatic plants in the removal process. Seeded MS2 bacteriophage and seeded poliovirus were removed more efficiently than were the indigenous bacteriophages, with less than 0.2% of MS2 and 0.1% of the poliovirus surviving flow at the same hydraulic application rate. The decay rate (k) of MS2 in a stagnant wetlands (k = 0.012 to 0.028 h-1) was lower than that for flowing systems (k = 0.44 to 0.052 h-1), reflecting the enhanced capacity for filtration or adsorption of viruses by the root-substrate complex (and associated biofilm).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3034154 TI - Physicochemical stability and inactivation of human and simian rotaviruses. AB - The effects of various physical and chemical treatments on the stability of a human serotype 1 rotavirus and simian agent 11 (SA11) were compared by using a fluorescence focus assay. The infectivity of both strains was retained after storage at room temperature for 14 days, 4 degree C for 22 days, and -20 degree C for 32 days; lyophilization; and treatment at pH 3 to 11. Both viruses were inactivated at pH 12, as was the human virus at pH 2, although this pH resulted in only partial inactivation of SA11. The human virus also appeared to be more sensitive than SA11 to the action of ether and chloroform. The infectivity of both viruses was lost after UV irradiation for 15 min and after treatment with 8% formaldehyde for 5 min, 70% (vol/vol) ethanol for 30 min, and 2% lysol, 2% phenol, and 1% H2O2 for 1 h each. PMID- 3034156 TI - Comparison of methods for rotavirus detection in water and results of a survey of Jerusalem wastewater. AB - Methods for the detection of viable rotaviruses and rotavirus antigen in water were developed and compared. The methods included laboratory-developed enzyme linked immunosorbent assays (ELISAs) with chromogenic and luminescent substrates, commercial Rotazyme and Enzygnost ELISAs, and an indirect immunofluorescent assay. Of the methods tested, the immunofluorescent assay and the Enzygnost ELISA were the most sensitive for the simian rotavirus SA-11. All of the methods were positive for human rotavirus from clinical specimens. Seeded SA-11 rotavirus was concentrated from water by absorption to and elution from Zeta Plus filters followed by organic flocculation. Interference with the assays by components of the wastewater concentrates was minimal for the ELISAs, although the undiluted organic flocs were cytotoxic for the immunofluorescent assay. A survey of Jerusalem wastewater was carried out over the course of 1 year, and samples were assayed for rotaviruses and enteroviruses. Although enteroviruses were found in almost all of the samples, all samples were negative for rotaviruses. The concentration of rotaviruses in the wastewater was thus below the detection limit of the method used. PMID- 3034157 TI - Synthesis and biological activity of 1 alpha,23,25,26-tetrahydroxyvitamin D3. AB - The metabolic pathway from 1 alpha,25-dihydroxyvitamin D3 [1 alpha,25-(OH)2D3] to 1 alpha,25-dihydroxyvitamin D3-26,23-lactone includes the formation of 1 alpha,23,25-26-tetrahydroxyvitamin D3 [1 alpha,23,25,26-(OH)4D3]. The aim of the current study was to explore the as yet unknown biological properties of this vitamin D3 sterol. The four diastereoisomers of 1 alpha,23,25,26-(OH)4D3 were chemically synthesized. They were compared to 1 alpha,25-(OH)2D3 in terms of their affinity for the chick intestinal 1 alpha,25-(OH)2D3 receptor and their biologic activity in vivo (stimulation of intestinal calcium absorption and mobilization of calcium from bone in vitamin D-deficient rats). The 1,25-(OH)2D3 receptor binding affinities of 1 alpha,23(R)25(R)26-(OH)4D3, 1 alpha,23(S)25(S)26 (OH)4 D3, 1 alpha,23(S)25(R)26-(OH)4D3, and 1 alpha,23(R)25(S)26-(OH)4D3 were 11, 100, 216, and 443 times weaker than the binding affinity of 1 alpha,25-(OH)2D3, respectively. Compared to 1 alpha,25-(OH)2D3, the relative capacities of the 1 alpha,23,25,26-(OH)4D3 compounds to stimulate intestinal calcium absorption were 1/4 for 1 alpha,23(R)25(R)26-(OH)4D3; 1/19 for 1 alpha,23(S)25(S)26-(OH)4D3; 1/90 for 1 alpha,23(S)25(R)26-(OH)4D3; and 1/136 for 1 alpha,23(R)25(S)26-(OH)4D3. Maximal stimulation of intestinal calcium transport occurred 8 h after administration of vitamin D3 metabolites. Mobilization of calcium from bone was quantitated by serum calcium concentration measurements. The activities of 1 alpha,23(R)25(R)26-(OH)4D3, 1 alpha,23(S)25(S)26-(OH)4D3, 1 alpha,23(S)25(R)26 (OH)4D3, and 1 alpha,23(R)25(S)26-(OH)4D3 to increase serum calcium were estimated to be 4, 13, 43, and 69 times weaker than that of 1 alpha,25-(OH)2D3, respectively. These results illustrate the stereospecificity of the chicken intestine 1 alpha,25-(OH)2D3 receptor for binding of 1 alpha,23,25,26-(OH)4D3 and suggest that the 1 alpha,23,25,26-(OH)4D3 exerts its biological activity in the rat through an interaction with 1,25-(OH)2D3 receptors. In summary, the 1 alpha,23,25,26-(OH)4D3 had a markedly lower biological activity than 1 alpha,25 (OH)2D3. PMID- 3034158 TI - Substrate specificity of choline kinase. AB - The substrate specificity of choline kinase (ATP:choline phosphotransferase, EC 2.7.1.32) from brewer's yeast has been examined using multiple analogs of choline, most of which have been reported to be a substrate of one or another choline-using system from other sources. In contrast to many such systems, choline kinase from brewer's yeast has been found to have relatively stringent and straight-forward structural requirements for its substrates. It is hypothesized that there are at least four points of interaction of the substrate with the enzyme--one for the hydroxyalkyl side chain and one for each of the three substituents on the quaternary nitrogen. Of the latter, one site seems relatively more sterically hindered than the other two. Short, single or double alkyl substitutions on the quaternary nitrogen are possible without a large loss of substrate capacity of the analog. Thus N,N-dimethyl-N-propylethanolamine had a relative Vmax of 116% and a relative Vmax 96% that of choline and a Km of 68 +/- 15 microM [nearly four times that of choline itself (18 microM)]. However, N butyl-N,N-dimethylethanolamine and N,N,N-triethylethanolamine were very poor substrates. Analogs with substituents on the quaternary nitrogen of longer chain length were without activity as were aromatic derivatives. None of the bisquaternary compounds of the general structure HOCH2CH2N+(CH3)2-(CH2)n N+(CH3)2CH2CH2OH (n = 2-10) showed any substrate capacity, as well. Restrictions on the hydroxyethyl side chain were also severe. One additional methylene group in this chain greatly reduced substrate capacity of the analog and two additional ones eliminated it entirely, as did almost any substituent on the beta carbon. A single (but not a double) substituent on the alpha carbon was moderately tolerated, however. Thus alpha-methylcholine and N-methyl-2 hydroxymethylpiperidine were substrates (although the latter one was a poor one) but beta-methylcholine and N-methyl-3-hydroxypiperidine were not. Such information may be of use toward designing cholinergic probes targeting specific enzyme or metabolic functions. PMID- 3034159 TI - Metabolism of nicotinamide mononucleotide in beef liver. AB - Nicotinamide mononucleotide (NMN) is not only an intermediate for the biosynthesis but also a degradation product of pyridine cofactors in animal tissues. Among the animal tissues tested, the highest NMN catabolizing activity was detected in beef liver (5.6 mumol/min/g tissue). This activity was 16 times higher than the NAD hydrolysis catalyzed by the liver NAD glycohydrolase. As a result of enzymatic analysis of the NMN splitting process, two types of enzyme responsible for this catabolism were partially purified and identified as a membrane-bound 5'-nucleotidase and a cytoplasmic nicotinamide riboside (NR) phosphorylase. No specific NMN glycohydrolase could be found in contrast to results observed in bacterial systems. The 5'-nucleotidase and NR phosphorylase constitute an obligatory process of the pyridine nucleotide cycle. The dephosphorylation and phosphorolysis catalyzed suggest that these enzymes could serve as an important mechanism for salvaging the ribose and nicotinamide moieties of NMN and pyridine nucleotides in the cell and a process that could be regulated at the mononucleotide level by this "NMN cycle" rather than by a NAD glycohydrolase cycle. In addition to the enzymatic properties of these enzymes, a regulatory mechanism by nucleotides such as ATP was also demonstrated. PMID- 3034160 TI - Iron-dependent uptake of ascorbate into isolated microsomes. AB - A preliminary study (J.M. Mata, R. Assad, and B. Peterkofsky (1981) Arch. Biochem. Biophys. 206, 93-104) suggested that chick embryo limb bone microsomes took up and concentrated [14C]ascorbate in the presence of cofactors for prolyl hydroxylase. In the present study, we found that the apparent Km for ascorbate in the hydroxylation of intracisternal unhydroxylated procollagen by endogenous prolyl hydroxylase was approximately an order of magnitude less than the value obtained when enzyme solubilized from microsomes was used with an exogenous substrate. These results are compatible with a concentrative uptake of ascorbate into microsomes. The uptake of [14C]ascorbate into microsomes was confirmed and it required only iron, in either the ferrous or ferric form, and was time and temperature dependent, proportional to microsome concentration, and substrate saturable at 2-3 mM ascorbate. Iron-dependent ascorbate uptake also was observed with L-929 cell microsomes. [14C]Ascorbate seemed to be taken up without prior oxidation, since only unlabeled ascorbate, and not dehydroascorbate, competed for uptake into limb bone microsomes. A functional requirement for Fe2+ in ascorbate transport was demonstrated using the intracisternal proline hydroxylating system. L-929 cell microsomes were preincubated with ascorbate with or without the metal and then external ascorbate was oxidized to inactive dehydroascorbate using ascorbic acid oxidase, which cannot penetrate the microsomal membrane. Samples which did not receive iron during the preincubation received it, along with other requirements for prolyl hydroxylase, in a final incubation to measure hydroxylation. Significant hydroxylation was obtained only in samples incubated with iron prior to oxidase treatment, consistent with the conclusion that an iron dependent process was required to translocate ascorbate and protect it from the oxidase. PMID- 3034161 TI - The c-type cytochromes of the gram-positive bacterium Bacillus licheniformis. AB - The release of soluble c-type cytochromes from cells of the gram-positive bacterium Bacillus licheniformis was effected by treatment with lysolytic buffer. After further purification three different c-type cytochromes designated c-551, c 552, and c-554 were isolated. Oxidized and reduced spectra, molecular weight, isoelectric point, and amino acid compositions are reported for each of these monoheme proteins. PMID- 3034162 TI - Superoxide removal and radiation protection in bacteria. AB - Previous work with procaryotic cells has identified one kind of lethal damage from ionizing radiation which occurs only within a specific range of low O2 concentrations, about 10(-6) to 10(-4) M. Within this range, protection can occur in three ways: through the enzymatic decomposition of hydrogen peroxide (H2O2) by added catalase, through the enzymatic degradation of superoxide anion radicals (.O2-) by added superoxide dismutase (SOD), and through scavenging hydroxyl radicals (.OH) by various additives. These results indicate that three radiolytic products, H2O2, .OH, and .O2- (and/or the conjugate acid, the perhydroxyl radical, .HO2) are involved in this single kind of radiation-induced damage. Although the radiolytic productions of H2O2 and .O2- are strongly enhanced in higher O2 concentrations, neither enzyme protects when these air-equilibrated bacteria are irradiated. These experiments address this apparent contradiction and focus on the specific issue of why the addition of SOD protects at low but not at high O2 concentrations. We propose that, at a given O2 concentration, .O2- (and/or .HO2) may either react (with some cellular component?) to cause damage or react (with itself) to form hydrogen peroxide (H2O2). The specific O2 concentration during irradiation would determine the relative rates of these competing reactions and therefore the O2 concentration itself would establish whether or not we will observe damage from .O2-. PMID- 3034163 TI - Photoaffinity labeling of opioid receptor of rat brain membranes with 125I(D Ala2, p-N3-Phe4-Met5)enkephalin. AB - A photoreactive (D-Ala2, p-N3-Phe4-Met5)enkephalin derivative was prepared, iodinated with carrier-free 125I, and then purified by high-performance liquid chromatography. The purified radioactive photoprobe was monoiodinated at the amino terminal tyrosine residue. This radioactive photoprobe was used to photoaffinity label membranes prepared from the rat brain (minus cerebellum) and the spinal cord. The photolabeled membranes were analyzed by sodium dodecyl sulfate gel electrophoresis. A 46,000-Da protein was specifically photolabeled in these membrane preparations. The photolabeling of this protein was inhibited by peptides related to enkephalin but not by unrelated substance P or gastrin tetrapeptide. A concentration-dependent inhibition of the photolabeling of the 46,000-Da protein was observed in the presence of competing ligands specific for the mu-, delta-, and kappa-opioid receptors. These data demonstrate that the radioactive photoprobe labels the mu-, delta-, and kappa-opioid receptors. Although there is no evidence available to show that the 46,000-Da protein is identical in all the cases, our data strongly suggest that it is a binding protein common to all of the opioid receptor subtypes. PMID- 3034164 TI - Pyrimidine nucleoside monophosphate kinase from rat bone marrow cells: a kinetic analysis of the reaction mechanism. AB - A kinetic analysis of the reaction mechanism of pyrimidine nucleoside monophosphate kinase was carried out with a highly purified enzyme preparation from rat bone marrow cells. The results of initial rate and product inhibition studies provided insight into the mode of action of the enzyme. The data support the views that the reaction mechanism is sequential and nonequilibrium in nature. Substrates bind to the enzyme in a random order. Substrate binding is cooperative. That is, the binding of the first substrate facilitates the binding of the second substrate. UMP can bind to the purine site on the enzyme, resulting in substrate inhibition. Product inhibition can result from the binding of UDP to either the pyrimidine or purine site, or from the binding of ADP to the purine site. PMID- 3034166 TI - [Transplantation in cancer therapy]. AB - In 1978, the new immunosuppressant, cyclosporin A, was accepted in clinical transplantation by Calne. Cyclosporin A eventually improved the results obtained after transplantation, after which it was widely accepted all over the world. Transplantation of several organs can be accepted as an effective form of treatment for patients with end-stage disease of each organ. Recently, transplantation has proved to be an excellent therapeutic method for cancer, especially in primary liver malignancy. Starzl has obtained good results in cases of fibrolamellar hepatomas. However, conservative surgical treatment should be used, whenever possible. Today, patients with unresectable liver tumor can expect long-term survival and even cure after liver transplantation with adjuvant therapeutic methods. Naturally, organ replacement is the most extensive surgical procedure for cancer treatment. However, early recurrence of tumors has often been observed following this therapy, possibly due to suppression. Adjuvant chemotherapy will thus be needed to improve the results of transplantation for malignancy. PMID- 3034165 TI - Superoxide radical initiates the autoxidation of dihydroxyacetone. AB - The aerobic xanthine oxidase reaction causes the cooxidation of dihydroxyacetone in a process which is strongly inhibited by superoxide dismutase but not by catalase, HO X scavengers, or iron-inactivating chelating agents. Several molecules of the sugar can be oxidized per O2- introduced. A free radical chain mechanism, in which O2- acts both as an initiator and as a chain propagator, is proposed. Simple sugars capable of tautomerizing to enediols may now be added to the list of biologically relevant targets for O2-. PMID- 3034168 TI - [Tumor necrosis factor]. AB - Tumor necrosis factor (TNF) is cytokine derived from macrophage and shows much promise for application in cancer therapy because of its marked antitumor effects and its high specificity to tumors. Recently, the gene encoding human TNF was expressed in E. coli and the recombinant TNF was purified to homogeneity by ion exchange chromatography, affinity chromatography, and gel filtration. Clinical study of rH-TNF has been launched because human recombinant TNF can be produced on a large scale. In spite of notable antitumor effects, little is known concerning the mechanism of action of cytotoxic activity. In this article, the mechanism of action of rH-TNF against tumors in vitro and in vivo is reviewed. PMID- 3034167 TI - [The role of cytokines in cancer therapy]. AB - A variety of normal tissue or malignant cells can produce and/or release various biologically active substances (hormone-like mediators) now collectively called cytokines. Because immunological and non-immunological responses of malignant cells were modified by many of them, some cytokines have been employed as so called Biological Response Modifiers (BRM) in the treatment of cancers in animals and humans. This overview discussed a few of the difficulties, probably inherent in cytokine therapy, that have already been encountered in early clinical trials as well as some of those that can be anticipated in future work. These include unexpected and undesirable reactions due to the systemic administration in relatively large amounts of a cytokine that is, under physiological conditions, supposed to act as a paracrine and/or autocrine among cells located within a limited distance. Even a pure recombinant preparation of a cytokine is now known to affect multiple target cells if they are accessible to it. Furthermore, this kind of therapy may sometimes be little more than a shot in the dark, since the physiological balance (homeostasis) among many of the cytokines present or produced in a host receiving a large quantity of exogenous cytokines is not well understood. Making the situation still more complicated, many types of tumor cells are known to release some of these cytokines spontaneously. Many challenging problems remain to be solved before we can confidently prescribe a cocktail of cytokines precisely suitable for a given patient according to the individual's in vivo cytokine profile. Nevertheless, in spite of all these reservations, cytokine therapy has been too frequently beneficial to be allowed to be discouraged. "Out of this nettle, danger, we pluck this flower, safety". PMID- 3034169 TI - [Prevention of hepatic metastases in rabbits by administration of an oily anticancer agent into the portal vein]. AB - We studied one kind of prophylactic chemotherapy against hepatic metastases. The therapy was carried out with a lymphographic oily contrast medium. Lipiodol, and a high-molecular-weight anticancer agent known as SMANCS. SMANCS was dissolved in Lipiodol by sonication (SMANCS/Lipiodol, 1 mg of SMANCS in 1 ml of Lipiodol). SMANCS/Lipiodol, administered into the portal vein, remained for a long time in the portal vein and was eliminated gradually through the bile and urine. SMANCS/Lipiodol (0.4ml/kg) was injected into the mesenteric vein in rabbits, which were then inoculated with the highly malignant carcinoma VX-2. Rabbits injected with SMANCS/Lipiodol before inoculation had significantly fewer hepatic metastases than the control 12 days later (P less than 0.001). Survival was significantly longer (P less than 0.005; 36.0 +/- 7.7 days) with SMANCS/Lipiodol before inoculation than without treatment (23.5 +/- 3.0 days). Hepatic metastases might thus be prevented by portal administration of an appropriate oily anticancer agent. PMID- 3034170 TI - [A phase I study of carboplatin]. AB - A phase I study of carboplatin was conducted using a single dose schedule. Escalating doses of 200, 300, 400 and 500mg/m2 were administered without hydration up to a total of 21 cycles in 18 patients with various solid tumors. A dose-limiting factor was thrombocytopenia, and leukopenia was also dose-related. A major clinical toxicity was gastrointestinal toxicity, while nephrotoxicity was extremely mild. The optimal dose for phase II trials was judged to be 300mg/m2 q 4w in poor-risk patients and 400mg/m2 q 4w in good-risk patients, respectively. Pharmacokinetics were studied in 11 patients. PMID- 3034171 TI - Hairy leukoplakia. A new disease of the oral mucosa. PMID- 3034173 TI - An enlarging tumor of the foot. Eccrine poroma. PMID- 3034172 TI - Oral hairy leukoplakia. A distinctive marker of human T-cell lymphotropic virus type III (HTLV-III) infection. AB - Oral hairy leukoplakia (HL) is a newly described lesion occurring principally on the lateral borders of the tongue in immunosuppressed homosexual men infected with human T-cell lymphotropic virus type III (HTLV-III). Clinically, HL appears as a slightly raised, poorly demarcated lesion with a corrugated or "hairy" surface. Histologically, the lesion is characterized by keratin projections on the surface (which often resemble hairs), parakeratosis, and acanthosis. In addition, large pale-staining cells with pyknotic nuclei are seen in the upper stratum malpighii, which appear similar to the koilocytes described in uterine condylomata. Candida organisms are frequently observed on the lesion surface. Little, if any, subepithelial inflammation is present. Human papillomavirus and Epstein-Barr virus have been identified in biopsy specimens from lesions of oral HL. The association of this lesion in patients with HTLV-III infection has been established. We saw a patient with HTLV-III infection and HL, in whom the immunochemical and ultrastructural findings revealed the presence of a mixed viral infection. Because oral HL may be of diagnostic value as an early indicator of HTLV-III infection, awareness of its characteristic clinical, histologic, immunochemical, and ultrastructural features is important. PMID- 3034174 TI - [Malignant mixed mesodermal tumor of the bladder. Presentation of a case]. PMID- 3034175 TI - The ubiquitous fibroblast. Multiple oncogenic potentials with illustrative cases. AB - The wide range of oncogenic proliferative potentials of the fibroblast is demonstrated with a series of eight patients. Diagnoses included infantile digital fibromatosis, "aggressive fibromatosis," aggressive fibromatosis progressing to poorly differentiated sarcoma, infantile myofibromatosis, recurrent desmoid tumor, fibrosarcoma arising in a keloid, dermatofibrosarcoma protuberans, and malignant fibrous histiocytoma of left atrium. Still other types of fibroblastic tumefactions might have been included. Oncogenic factors that may have been operative in the causation of the lesions presented include: genetic factors, sex-linked factors, hormonal factors, numerous growth factors, and certain viruses, especially retroviruses. Certain fibromatoses in children are commonly self-limited and need only be monitored carefully as the process regresses. Aggressive fibromatosis, on the other hand, can prove fatal if the lesion is not completely resected with a wide margin and, occasionally, the process may become frankly malignant, with metastases. The standard triad of excisional surgery, radiotherapy, and chemotherapy has been used to treat frankly malignant fibrous tumors with variable results. PMID- 3034176 TI - Mediastinal tracheostomy. AB - Upper airway obstruction in primary or recurrent carcinomas of the head and neck extending into the mediastinum may demand surgical intervention despite severe technical difficulties in patients with tumors previously considered inoperable. In fact, many of these tumors may be operable and some perhaps curable. A technique has been developed based in part on our experience with previously described procedures. A preliminary sternal split is used to demonstrate the extent of the mediastinal involvement as well as to provide enhanced exposure and proximal control of the great vessels. The pectoralis major muscle is used with a generous flap of overlying skin comprising nearly half of the anterior portion of the chest. A tracheostomy is then created in a fashion similar to the placement of a cardiac valvular prosthesis by creating a circular defect in the pectoralis major flap and suturing it to the tracheal remnant. This technique offers a reasonably safe and reliable means of creating a low anterior mediastinal tracheostomy for tumors previously considered inoperable. The preliminary sternal split makes the procedure safer and easier to perform, and the use of a very large pectoralis major island flap allows for reliable closure of the resulting mediastinal and sternal defects. PMID- 3034178 TI - Unusual neurotoxicity associated with amiodarone therapy. AB - One hundred two patients with recurrent, drug-refractory tachyarrhythmias were treated with amiodarone for nine +/- eight months (mean +/- SD) (range, one to 50 months). Forty-five patients exhibited some form of neurotoxic reaction that was severe enough in nine patients to require discontinuation of treatment or reduction in dosage of the drug. The most frequent neurotoxic findings were tremor (44 patients), peripheral neuropathy (ten patients), and ataxia (seven patients). Five patients developed unusual neurotoxic manifestations: brainstem dysfunction characterized by downbeat nystagmus, hemisensory loss and ataxia, severe dyskinesia, jaw tremor, and proximal myopathy. Neurophysiologic studies revealed varying degrees of predominantly demyelinating peripheral neuropathy. Neurotoxic symptoms improved after discontinuing treatment or decreasing the dosage of the drug. Age of the patient and total cumulative dose did not seem to be risk factors for development of neurotoxicity. These neurotoxic findings suggest that amiodarone-induced neurotoxic reactions are not only confined to the peripheral nervous system, but also that parts of the central nervous system (eg, basal ganglia, brain stem, or their connections) may also be involved. PMID- 3034177 TI - Marked hyperbilirubinemia in infectious mononucleosis. Analysis of laboratory data in seven patients. AB - While mild to moderate hepatic dysfunction is commonly encountered in infectious mononucleosis induced by Epstein-Barr virus (EBV), clinical jaundice with high bilirubin levels (greater than or equal to 6.0 mg/dL [greater than or equal to 103 mumol/L] is only occasionally encountered. In this study, seven patients with primary EBV infections had peak bilirubin levels of 10.2 to 23.0 mg/dL (174 to 393 mumol/L) and, for the most part, presented initial diagnostic problems. Complications included the virus-associated hemophagocytic syndrome and acute respiratory distress syndrome in one patient and transient renal failure in another. The laboratory data suggested that a combination of hemolysis and viral induced cholestasis was responsible for the intense hyperbilirubinemia in at least five patients. Physicians should be aware that marked hyperbilirubinemia can occur with EBV-induced infectious mononucleosis and, thereby, obviate the need for costly diagnostic laboratory tests and, occasionally, invasive procedures. PMID- 3034179 TI - Occurrence of renal tubular dysfunction in lupus nephritis. AB - We prospectively evaluated 30 patients who presented with active systemic lupus erythematosus (SLE) for the presence of tubular abnormalities. All patients fulfilled the American Rheumatology Association criteria for SLE. When appropriate, a renal biopsy was performed. Of the 30 patients studied, 12 had no abnormal tubular study results, whereas 18 patients had some form of defect in the handling of potassium, sodium, or hydrogen ions. Eight patients had distal renal tubular acidosis (dRTA) due to an isolated proton secretory defect. Five had dRTA of the gradient or acid back-leak type. Two had an unresponsive voltage dependent form of dRTA; one had a responsive voltage-dependent form of dRTA. One individual had hyporeninemic hypoaldosteronism and one had dRTA plus hypoaldosteronism. Clinically, patients with the abnormal tubular study results more often presented with nephritis or nephrotic sediment, peripheral edema, or anemia. Renal biopsies failed to demonstrate any difference in glomerular histologic findings and calculated activity, chronicity, or interstitial indexes. We conclude that SLE may be associated with a variety of tubular defects. PMID- 3034180 TI - Use of the glucose clamp technique for confirmation of insulinoma autonomous hyperinsulinism. AB - The diagnosis of insulinoma on the basis of persistent hypoglycemia requires further confirmation. The insulin suppression test has been used to support this diagnosis prior to surgical intervention. In this study the euglycemic clamp technique was used to compare five control volunteers with four hypoglycemic patients with suspected insulinoma. Insulin was infused over successive two-hour periods at 2, 4, and 8 mU/kg/min. Plasma glucose levels were clamped at 80 mg/dL (4.4 mmol/L) using an artificial pancreas. High insulin levels were measured in all subjects, ranging from 225 +/- 30 microU/mL (1614 +/- 215 pmol/L) to 1018 +/- 239 microU/mL (7304 +/- 1714 pmol/L). Levels of C peptide fell to 0.1 ng/mL (0.028 nmol/L) in control subjects but remained at high levels in the patients. Insulinoma was confirmed on laparotomy in all four patients. In two patients tested after removal of the tumor the results were found to have returned to normal. PMID- 3034181 TI - Prolonged suppression of a corticotropin-producing bronchial carcinoid by oral bromocriptine. AB - The addition of orally administered bromocriptine mesylate to cyproheptadine hydrochloride therapy completely normalized urinary-free cortisol levels for three months in a 21-year-old woman with Cushing's syndrome in whom results from standard dexamethasone suppression and metyrapone stimulation tests as well as baseline corticotropin levels were originally compatible with a diagnosis of an occult pituitary adenoma. When transsphenoidal exploration of the sella turcica was unsuccessful and hypercortisolism persisted, the source of corticotropin was discovered using petrosal sinus and venal caval catheterization. A 1 X 1.5-cm carcinoid tumor of the lung was identified and removed, thereby correcting the hypercortisolism. The tumor was demonstrated by immunoperoxidase staining to contain corticotropin. Orally administered bromocriptine, with or without cyproheptadine therapy, may be useful in the palliative treatment of some patients with carcinoid or other ectopic corticotropin-producing tumors. We postulate that bromocriptine therapy acted directly on carcinoid tumor cells to directly inhibit corticotropin production by a dopaminergic mechanism. PMID- 3034182 TI - Technetium Tc 99m uptake in postpartum thyrotoxicosis. PMID- 3034183 TI - [Extrahepatic hepatocellular cancer]. PMID- 3034184 TI - [Subungual glomus tumors. Apropos of 2 cases]. AB - Glomus tumours are uncommon, though not exceptional, benign growths developed from the constituents of the "neuromyoarterial glomus" initially described by Masson. Two cases of subungual glomus tumours of the hand in middle-aged women are reported. In spite of characteristic symptoms, with paroxysmal episodes of pain triggered off by cold and the most minute traumas, and of the finding in both cases of a violine++ tumour under the nail, the condition was belatedly diagnosed (7 and 27 years respectively after the onset of symptoms). Diagnosis rested not so much on standard radiography, which showed erosion of the distal phalanx in both patients, as on arteriography of the hand which displayed a small vascular pool at the arterial stage. Tumoral excision was successful. PMID- 3034185 TI - Methylation sensitivity of the restriction enzymes FnuDII and AccII. AB - The restriction enzymes FnuDII and AccII are isoschizomers of the DNA sequence 5' CGCG-3'. We have determined that 5-methylcytidine at either cytidine position in this recognition sequence inhibits DNA cleavage by FnuDII and AccII. A third isoschizomer, ThaI was previously shown to exhibit an identical methylation sensitivity. It is remarkable that 3 restriction enzymes derived from diverse microbiological sources exhibit this identical methylation sensitivity. PMID- 3034186 TI - Construction of a transposon containing a gene for polygalacturonate trans eliminase from Klebsiella oxytoca. AB - A DNA fragment containing a Klebsiella oxytoca gene for polygalacturonate trans eliminase was cloned into the kanamycin resistance transposon Tn5. This new transposon, designated Tn5-Pga+, had a transposition frequency of 1 X 10(-6). The broad host range plasmid pR751::Tn5-Pga+ was conjugally transferred to a variety of genetic backgrounds. The ability to degrade polygalacturonate was expressed in Aeromonas hydrophila, Alcaligenes eutrophus, Azotomonas insolita, Escherichia coli, Pseudomonas putida and Rhodopseudomonas sphaeroides, but not in Zymomonas mobilis. PMID- 3034187 TI - [Dietetic control of blood cholesterol and the prevention of atheroma]. PMID- 3034188 TI - Brain gamma-aminobutyric acid abnormality in tardive dyskinesia. Reduction in cerebrospinal fluid GABA levels and therapeutic response to GABA agonist treatment. AB - A double-blind, placebo-controlled trial of gamma-vinyl gamma-aminobutyric acid (GVG) and 4,5,6,7-tetrahydroisoxazolo-(5,4-c) pyridine-3-ol (THIP) was carried out in drug-free schizophrenic patients with tardive dyskinesia. A significant decrease in dyskinetic symptoms occurred with the administration of GVG, associated with a twofold increase in cerebrospinal fluid levels of GABA; THIP produced a more moderate, yet consistent decrease in the involuntary movements. A pathophysiologic role for gamma-aminobutyric acid (GABA)-mediated neuronal transmission in tardive dyskinesia was explored by analyzing cerebrospinal fluid GABA concentrations in drug-free schizophrenic patients with and without tardive dyskinesia. A significant reduction in cerebrospinal fluid levels of GABA was observed in the dyskinetic schizophrenics compared with the nondyskinetic controls. These data compliment a growing body of experimental evidence suggesting a critical role for GABA-ergic neurons in the pathophysiology of tardive dyskinesia. PMID- 3034189 TI - Giant cell pneumonia caused by parainfluenza type 3 in a patient with acute myelomonocytic leukemia. AB - A 4 1/2-year-old boy with acute myelomonocytic leukemia developed fever, neutropenia, and a prolonged respiratory illness while receiving maintenance chemotherapy. An open lung biopsy specimen demonstrated a giant cell pneumonia with intracytoplasmic and probable intranuclear viral inclusions of the paramyxovirus type. Serologic studies demonstrated convincing evidence of a parainfluenza type 3 infection. Although parainfluenza type 3-induced giant cell pneumonia has been reported in infants with the severe combined immunodeficiency syndrome, to our knowledge, this is the first reported case of this complication in a patient with leukemia. PMID- 3034190 TI - [CNS-active 4-phenylpyranes: 9-phenyl-9-piperidino-3-oxa-7 azabicyclo[3.3.1]nonane]. PMID- 3034191 TI - [Oligopeptides of beta-carboline-3-carboxylic acid--synthesis and affinity for benzodiazepine receptors]. PMID- 3034192 TI - [Synthesis and antiviral activity of quinoxalinedione 2,3(1H, 4H) derivatives]. PMID- 3034193 TI - Gastric partitioning complicated by peripheral neuropathy with lumbosacral plexopathy. AB - Gastric bypass and partitioning are the two surgical procedures most commonly used in the treatment of morbid obesity. They are, however, not without their postoperative complications. These include acute and chronic problems such as wound infection, gastric leak, obstruction, embolism, and neurologic sequelae. Many studies have mentioned the frequent occurrence of polyneuropathy in the postgastrectomy state. This report describes a 38-year-old patient who developed an asymmetric peripheral neuropathy with lumbosacral plexus involvement following gastric bypass surgery for morbid obesity. PMID- 3034194 TI - Intraoperative implantation radiation therapy plus lumpectomy for carcinoma of the breast. AB - The preponderance of evidence indicates that lumpectomy plus radiation therapy produces the same survival as modified radical mastectomy in patients with early breast cancer and further suggests that the local failure rate may be lower in the patients who undergo irradiation. In this series, patients were treated with immediate implantation of the tumor bed with iridium Ir 192. There have been two recurrences in the breast from 110 breast cancers (44% had a clinical stage greater than T1, and 41% had axillary-node involvement) in 107 patients followed up for four to 52 months (25.8 +/- 13.3 months [mean +/- SD]). These preliminary data suggest that local treatment failure can be minimized by aggressive, immediate intraoperative implantation of the tumor bed with iridium Ir 192. PMID- 3034196 TI - Calmodulin-mediated cadmium inhibition of phosphodiesterase activity, in vitro. AB - Ion-stripped bovine brain calmodulin (CaM) binds 4 moles Cd2+ as well as 4 moles Ca2+ per mole protein, with similar affinity; in the presence of 1 mM Mg2+ the molar binding ratio of CaM for Ca2+ decreased to 3, the apparent K0.5 for Ca2+ nearly doubled, but the binding characteristics of CaM for Cd2+ were not changed. Saturating concentrations Ca2+ did not affect the molar binding ratio of CaM for Cd2+, but increased the apparent K0.5 for Cd2+; vice versa, saturating concentrations Cd2+ decreased the molar binding ratio for Ca2+ to 2 without affecting the apparent K0.5 for Ca2+. CaM-independent phosphodiesterase (PDE) activity was inhibited at [Cd2+] greater than 10(-5) M. Cd2+-CaM as well as Ca2+ CaM activated PDE. However, the Cd2+-CaM complex is less effective than the Ca2+ CaM complex in stimulating CaM-dependent enzyme activities. Cd2+ inhibits Ca2+- and CaM-dependent PDE in a competitive way. Introduction of Cd2+ in a medium containing Ca2+ and CaM may, therefore, result in a reduction of CaM-dependent enzyme stimulation. By its interference with Ca2+- and CaM- dependent PDE activity, Cd2+ could upset the catabolic pathway of cellular cyclic nucleotide metabolism. PMID- 3034195 TI - Interaction of the pesticide chlordimeform with adrenergic receptors in mouse brain: an in vitro study. AB - Chlordimeform (N'(4-chloro-o-tolyl)-N, N-dimethylformamidine; CDM) is a formamidine insecticide acaricide whose major active metabolite is its N monomethyl analog, desmethylchlordimeform, (DCDM). While their pesticidal action in invertebrates appears to be related to activation of octopamine receptors, their mechanism of action in mammals has not been established. Because of similarities between octopamine and adrenergic receptors and suggestions of CDM and DCDM action on adrenoceptors, the in vitro interactions of CDM and DCDM with adrenoceptors were studied. In mouse brain membrane preparations CDM inhibited the binding of [3H]-clonidine to alpha 2- adrenoceptors and of [3H]-WB4101 to alpha 1-adrenoceptors with IC50 values of 18.2 and 87 microM, respectively. DCDM was a much more potent inhibitor, with IC50 values toward alpha 2-, and alpha 1 adrenoceptors of 44 nM and 1 microM, respectively. Both compounds were only weak inhibitors of the binding of [3H]-dihydroalprenolol to beta-adrenoceptors and of [3H]-quinuclidinyl benzilate to muscarinic receptors and were inactive toward benzodiazepines and gamma aminobutyric acid (GABAA) receptors. Inhibition of [3H] clonidine binding by both compounds was competitive, as indicated by a decreased receptor affinity without changes in receptor density. Interaction of CDM and DCDM with [3H]-WB4101 binding, on the other hand, was more complex, and not of the competitive type. These results show that CDM and its metabolite DCDM can interact directly in vitro with alpha-adrenergic receptors, suggesting that these receptors could mediate some of the effects of CDM and DCDM in vivo. PMID- 3034198 TI - Reactivity of monoclonal antibodies to proteins of a neurotropic bovine herpesvirus 1 (BHV-1) strain and to proteins of representative BHV-1 strains. AB - 15 monoclonal antibodies (McAbs) were induced with the neuropathogenic strain N 569 of bovine herpesvirus 1 type 3 (BHV-1.3). Nine of them could be shown by radioimmunoprecipitation assay to react with viral glycoproteins and two of these McAbs were able to neutralize strain N 569. The reactivity of these 15 monoclonals was compared with 11 monoclonal antibodies induced with a BHV-1.1 strain. The available monoclonal antibodies made it possible to characterize BHV 1.3 and to classify BHV-1 into three types, namely BHV-1.1, BHV-1.2 and BHV-1.3. This confirmed the results based upon restriction endonuclease analysis and viral protein patterns obtained earlier. The main antigenic differences of representative virus strains were found on two glycoproteins designated 3 and 12. Caprine herpesvirus 1, included in this study because of its serological relationship to BHV-1, differed fundamentally from BHV-1 on the grounds of McAb reactivity. PMID- 3034197 TI - The mode of assembly of alphavirus cores implies a mechanism for the disassembly of the cores in the early stages of infection. Brief review. PMID- 3034199 TI - Host-dependent temperature-sensitive growth of HVJ (Sendai virus) wild-type in rat glioma C 6 cells. AB - At non-permissive temperature viral specific RNA synthesis was not restricted in rat glioma (C 6) cells infected with HVJ (Sendai virus) wild-type. However, as has previously been shown (J Gen Virol [1984] 65: 639-643), the synthesis of M protein was reduced at non-permissive temperature, in contrast to the L, P, HN, Fo and NP proteins which were synthesized in comparable amounts at permissive and non-permissive temperatures. In this report we show additionally that viral nucleocapsids (NC), which consist of L, P and NP proteins, were formed within the infected cells at both temperatures. Hemagglutinin and neuraminidase activities were also detected in samples incubated at non-permissive temperature. By membrane immunofluorescence and cell-surface immunoprecipitation it was shown that migration of HN and Fo proteins to the cell surface occurred normally at non permissive temperature. Additionally, the L, P and NP proteins, which were associated with the plasma membrane isolated from the infected cells maintained at permissive temperature, were absent from the membrane of cells incubated at non-permissive temperature. These results suggest that NC and glycoproteins synthesized at non-permissive temperature could not assemble effectively at the plasma membrane because of a lack of M protein. Thus, the host-dependent ts lesion of HVJ in C 6 cells was considered to be mainly in M protein synthesis. PMID- 3034201 TI - Isolation and partial characterization of an orthopoxvirus from a California vole (Microtus californicus). Brief report. AB - A virus with the characteristics of an orthopox was isolated from a scab removed from the foot of a California vole. HAI antibody surveys indicated that the virus in enzootic in several different vole populations. PMID- 3034200 TI - The sequence of the coxsackievirus A 21 polymerase gene indicates a remarkably close relationship to the polioviruses. Brief report. AB - We have sequenced the coxsackievirus A 21 polymerase gene and 3' noncoding region. The sequence is 98 per cent homologous at the amino acid level to the three poliovirus serotypes. This is comparable to the relationship between polioviruses and indicates a recent evolutionary divergence of the viruses. PMID- 3034202 TI - Detection of bluetongue virus (BTV) antigen. PMID- 3034204 TI - Variation in the intracellular polypeptide profiles from different isolates of bovine virus diarrhoea virus. AB - Variation of the intracellular polypeptides induced in calf testis cells by 5 cloned isolates of bovine virus diarrhoea virus (BVDV) was examined. Three of the isolates were cytopathic (NADL, C 2415 and Pe 515 c) and two were non-cytopathic (C 1226 and Pe 515 nc) in these cells. The isolates Pe 515 c and Pe 515 nc were both isolated from an animal with clinical signs of mucosal disease. In cells infected with NADL, 8 virus specific proteins (vp 1 to vp 8) with molecular weights ranging from 120,000 (vp 1) to 23,000 (vp 8) were detected. Isolates C 2415 and Pe 515 c gave a similar array of polypeptides to NADL, but the 3 cytopathic isolates could be distinguished by the variation in the molecular weights of some of the proteins. The non-cytopathic isolates could also be distinguished from each other by this type of molecular variation; however, one feature that characterised these strains, when compared to the cytopathic isolates, was the absence of vp 2. Comparison of the polypeptides induced by Pe 515 c and Pe 515 nc showed that apart from the lack of vp 2 in the Pe 515 nc virus profile, the molecular weights of the other viral proteins were similar. This supports serological evidence that for mucosal disease to occur the pair of cytopathic and non-cytopathic viruses must be closely related. Four of the polypeptides induced by Pe 515 c were shown to be glycoproteins. PMID- 3034203 TI - Characterization of a second bovine rotavirus serotype. AB - Bovine rotavirus (BRV) V 1005 was characterized by two-way cross-neutralization tests as a second serotype of BRV. Virions and inner shell particles of 65 nm and 55 nm diameter respectively, and empty capsids of 65 nm and 55 nm diameter were separated by density gradient centrifugation. Three polypeptides of molecular weight 60,000, 36,000 and 28,000 (minor protein) could be identified in the outer shell of virions and in the larger empty capsids. Inner shell particles contained three polypeptides of molecular weight 105,000, 83,000 and 43,000. Both sizes of empty capsids showed two polypeptides of molecular weight 75,000 and 55,000 not found in virions. Pulse-labelling of infected cells revealed eight major and three minor intracellular viral polypeptides. Viral polypeptides synthesis started at about 6 hours p.i. and correlated in time with double-stranded RNA synthesis. As soon as viral polypeptide synthesis was detectable, newly synthesized viral polypeptides were incorporated into intracellular viral particles. Radioactive viral polypeptides appeared without a longer lag period in extracellular viruses from 6 hours p.i. onwards. PMID- 3034205 TI - Expression of the HSV specified major DNA-binding protein in virus infected RAJI and VERO cells. AB - We studied the expression of the Herpes Simplex virus type-2 (HSV-2) specified major DNA-binding protein (ICSP 11/12) in virus infected RAJI and VERO cells. Immunofluorescence staining by a specific ICSP 11/12 antiserum distinguished between different cell-virus interactions. ICSP 11/12 synthesized in the HSV-2 infected VERO cells showed dense homogenous nuclear staining early in the infection. During virus replication the pattern changed into a less dense, somewhat marginating staining. The corresponding protein synthesized during persistent HSV-2 infection in the RAJI cells showed granular cytoplasmic staining. Late in the infection a whole cell fluorescence was noted. In the absence of virus DNA-synthesis (PFA treated VERO cells) ICSP 11/12 showed dense homogenous or speckled nuclear pattern. The ICSP 11/12 protein extracted from the different virus-cell interactions also showed different elution from double stranded DNA. PMID- 3034207 TI - Beta interferon production in primed and unprimed cells infected with human cytomegalovirus. AB - Human cytomegalovirus induced beta interferon in cultures of human foreskin cells. The inhibitor was first released between 8 and 16 hours after infection, about 48 hours before progeny virus. In cultures infected with low concentrations of virus, interferon was produced as the infection spread, and then in amounts larger than expected. After infection with cytomegalovirus, cells which had been primed for 48 hours with purified beta interferon produced significantly more interferon than unprimed cells, and the interferon was produced earlier, between 2 and 8 hours after infection. CMV-induced interferon also was able to prime cells. The data suggest that the relatively large quantities of interferon detected in cultures infected with low concentrations of cytomegalovirus result from endogenous priming: those cells infected early first produce interferon which primes uninfected cells, then virus which induces the primed cells to produce interferon in relatively high concentrations. PMID- 3034206 TI - Cellular changes associated with persistent hepatitis A infection in vitro. AB - The rate of division, morphology and ultrastructure of BSC-1 cells, persistently infected with hepatitis A virus (HAV), were compared with uninfected cells for 60 days after splitting of the cells. Both control and infected cells showed a biphasic growth pattern marked firstly by increasing cell density and high mitotic rate (exponential phase) and then high constant cell density and little mitosis (stationary phase). Immunoperoxidase studies showed that hepatitis A antigen (HAAg) appeared as cytoplasmic granules approximately one third of the way through the exponential phase in infected cells. The percentage of cells with HAAg rose until the early stationary phase when virtually all cells contained antigen. Radioimmunoassay demonstrated an increase in HAAg per cell in the stationary phase. Radioimmunofocus assay and immune electron microscopy confirmed the presence of HAV in infected cells in the stationary phase. Thin sectioning electron microscopy showed cytoplasmic annulate lamellae in infected cells of both phases but not in control cells. PMID- 3034208 TI - Cultivation of avian rotaviruses in chicken lymphocytes and lymphoblastoid cell lines. AB - Avian rotavirus isolates were used to infect normal chicken spleen cells, lymphoblastoid T cell lines transformed by Marek's disease virus, an avian leukosis virus-transformed B cell line, and a reticuloendotheliosis virus transformed line, which is a pre-B, pre-T cell line. All five isolates tested were able to infect spleen cells and the three types of lymphoblastoid cell lines, suggesting that avian rotaviruses can infect both B and T cells. Splenic lymphocytes were considerably less susceptible to infection than chick kidney cells. Lymphoblastoid cell lines remained virus-positive during a 10-day culture period. Virus was neutralized by the addition of low dilutions of normal chicken serum and high dilutions of chicken anti-rotavirus serum. PMID- 3034209 TI - Variation in distribution of the three flavivirus-specified glycoproteins detected by immunofluorescence in infected Vero cells. AB - Indirect immunofluorescence with rabbit antisera was used to probe the intracellular locations of the antigens of envelope, prM (precursor to structural protein M) and the nonstructural glycoproteins NS 1 (formerly described as NV 3 or SCF) specified by the flaviviruses dengue-2 and Kunjin. Perinuclear staining in various types of foci was prominent for all antigens, and the distribution was influenced by whether cells were fixed with acetone or formaldehyde. Staining of Golgi-like masses or inclusions by anti-envelope sera occurred regularly and prominently in cells infected and stained with homologous anti-envelope antibodies; in the cross reactions, such staining was largely absent, especially in dengue-2 infected cells in which it was replaced by many small circular foci scattered throughout the cytoplasm. Anti-NS 1 also stained large perinuclear inclusions and small cytoplasmic foci, but the distribution of these was dissimilar to that observed with anti-envelope sera. Anti-prM appeared to contain a mixture of antibodies of different specificities, evident at different dilutions, possibly because of different cytoplasmic locations of prM and its cleavage products. All antisera produced small discontinuous foci on the plasma membrane of unfixed infected cells; antigens of NS 1 were sometimes prominent on the surface of acetone-fixed cells. PMID- 3034211 TI - Induced proteins in cells infected with pseudorabies virus. AB - New proteins appearing after infection of cultured L929 cells with pseudorabies virus (PRV) were analyzed by SDS polyacrylamide gel electrophoresis. Analysis was facilitated by using a virus-cell system with marked inhibition of host protein synthesis after infection. Infected cells were pulsed during successive two hour periods through the infectious cycle with 35S-methionine. Proteins were extracted with detergent and analyzed on high resolution reducing gels. Thirty-four protein bands were resolved on gels of different concentrations that varied from 7 to 15 percent. Calculated apparent molecular weights of the protein peaks were not dependent on gel concentration except for very large or small sized proteins. Eight glycoproteins were resolved after labeling with 14C-glucosamine. The time course of incorporation of label was used as a measure of protein synthesis allowing the grouping of proteins according to the time of maximal synthesis. Several proteins shifted in MW during the course of infection, indicating possible post-translational cleavage or other minor modification. PMID- 3034210 TI - Inhibition by monensin of human cytomegalovirus DNA replication. AB - Monensin, at concentrations which depended on the multiplicity of infection, was found to prevent DNA replication of human cytomegalovirus (HCMV) as well as production of viral progeny in human foreskin fibroblasts. The drug did not affect DNA replication of herpes simplex virus. Inhibition of consecutive HCMV DNA synthesis was also observed following delayed addition of the drug within 12 24 hours postinfection, but was fully reversible upon its removal. Viral replication proceeded, however, without impairment in cultures treated with monensin prior to infection. Induction of viral DNA polymerase activity was not impeded by the inhibitor. Analysis of protein- and glycoprotein synthesis revealed that monensin interfered with the production of a number of HCMV specific polypeptides. Furthermore, evidence was obtained that the drug may hinder intracellular transport of a 135 kd glycopolypeptide. PMID- 3034212 TI - A new human adenovirus of subgenus D: candidate adenovirus type 42. AB - A new serotype, candidate adenovirus type 42, a member of subgenus D, was isolated from the feces of a healthy child. It shared its hemagglutinin with adenovirus type 15, but had no relationship in neutralization to other human adenoviruses. It also had a distinct DNA restriction pattern. PMID- 3034213 TI - Susceptibility of chicken blood lymphoblasts and monocytes to infectious bursal disease virus (IBDV). AB - PHA-M stimulated lymphoblasts obtained from peripheral blood and separated from small lymphocytes by X 1 g velocity sedimentation, unstimulated blood lymphocytes, monocytes and cells isolated from the bursa of Fabricius of chickens, were infected in vitro by the pathogenic strain CU-1 of infectious bursal disease virus (IBDV). Six hours after infection 32.5 per cent of the bursal cells reacted immunocytologically with IBDV antiserum and had high infectivity titers in plaque assays. Separated lymphoblasts showed a marked lower degree of virus replication and only 2.5 per cent reacted positively when studied by immunocytology, while monocytes ranged between these two cell types with regard to both the degree of virus replication and the positive reaction with IBDV antiserum. Small lymphocytes, however, were found to be totally resistant to IBDV infection. When studied by electron microscopy, virus particles arranged in a crystalloid pattern could only be detected in bursal cells. The results of this study indicate that proliferating lymphoid cells at a certain stage of cellular differentiation are the target cells for IBDV, and that in infected chickens monocytes may play a role in the spreading of the virus. PMID- 3034214 TI - Chemically purified serum factor can induce both Epstein-Barr virus antigen synthesis and cell differentiation. AB - The effects of purified Epstein-Barr virus-inducing serum factor (EIF) on the lymphoblastoid cell line Raji latently infected with Epstein-Barr virus were studied. Activated serum factor was capable of both: cooperating with n-butyrate and/or TPA in inducing synthesis of EBV antigens and triggering cell differentiation towards plasma cell as determined by electron microscopy. It seems therefore that the same serum component was involved in the induction of both phenomena. PMID- 3034215 TI - Effect of recombinant human interferon gamma against human cytomegalovirus. AB - Escherichia coli-derived human interferon-gamma (rIFN-gamma) inhibited the replication of human cytomegalovirus (HCMV) synergistically when combined with IFN-alpha. The induction of HCMV DNA polymerase was inhibited in rIFN-gamma treated cells. It is suggested that the induction of 2-5 A synthetase does not play an important role in the anti-HCMV actions of IFNs. PMID- 3034216 TI - [Studies of the effect of active oxygen on the cytotoxicity of erythrocytes]. PMID- 3034217 TI - [Intranuclear inclusions in the cells of different tissues of the aging rat]. AB - Ultrastructure of neuronal and glial nuclei has been studied in various parts of the CNS, in parenchymatous cells of internal organs, in endotheliocytes and pericytes of their blood capillaries in intact rats--mature (6-8-month-old) and old (26-28-month-old), as well as at certain experimental influences. In all the cellular populations studied (most often in neurons), various in their structure, intranuclear inclusions (INI) have been revealed, that are considered as five main types. The number of INI increases sharply at ageing, that is especially noticeable under experimental conditions. Some INI are situated in morphologically preserved, actively working cells, being normal components of the nucleus, others--reflect profound rearrangements of nuclear proteins, disturbances in lipid metabolism up to irreversible destructive processes in the nucleus. PMID- 3034219 TI - Somatosensory evoked potentials: usefulness in cases of radiculopathy? PMID- 3034218 TI - [Enalapril in the treatment of severe congestive heart failure. Double-blind study, controlled with placebo]. PMID- 3034220 TI - Phosphorus magnetic resonance spectroscopy of partially blocked muscle glycolysis. An in vivo study of phosphoglycerate mutase deficiency. AB - In vivo phosphorus magnetic resonance spectroscopy was used to evaluate the changes in muscle bioenergetics in a patient with a partial glycolytic block. Phosphoglycerate mutase-deficient muscle showed the following evidence: Abnormal accumulation of sugar phosphates does occur, even when 6% enzyme activity is present. The elimination of sugar phosphates was faster than in complete glycolytic blocks. Mild intracellular acidosis occurred during ischemic exercise. The energy state was slightly low at rest but not during exercise. Postexercise recovery was mildly slowed. These findings suggest that phosphorus magnetic resonance spectroscopy can detect partial defects, as well as full glycolytic blocks, in muscle metabolism. PMID- 3034221 TI - Decrease of ocular pressure with oral metyrapone. A double-masked crossover trial. AB - To examine the effect of a decrease in plasma cortisol level on intraocular pressure, 14 subjects with elevated ocular pressure were given oral metyrapone tartrate (an inhibitor of adrenal cortisol biosynthesis) and a placebo in a double-masked, two-period, crossover trial. Ocular pressure and the adrenal response were monitored over a seven-hour period. The decrease in ocular pressure after metyrapone administration was significantly greater than that after placebo. The 14 subjects could be designated as responders or nonresponders by decline of ocular pressure after metyrapone administration compared with response after placebo. The responders could be further differentiated from the nonresponders by a smaller decrease in ocular pressure over time in response to the placebo and by a smaller decrease over time of plasma cortisol level in response to metyrapone. These results provide further evidence that plasma glucocorticoids may play a role in the regulation of ocular pressure. PMID- 3034222 TI - Ocular involvement associated with chronic Epstein-Barr virus disease. AB - Ocular involvement with acute Epstein-Barr virus infection is usually limited to a transient follicular conjunctivitis, although other lesions have been reported. Chronic Epstein-Barr virus infection has recently gained attention, but ocular manifestations have not been emphasized. We describe three patients with chronic infection with prominent ocular involvement. Bilateral uveitis was noted in all patients, ranging from an anterior uveitis that was responsive to steroids to a severe panuveitis with vitritis, cataract, optic disc swelling, and macular edema. In one patient, topical acyclovir ointment resulted in a substantial decrease in the inflammatory reaction when added to systemic acyclovir therapy. Another patient displayed a keratitis that resolved with topical steroid therapy. Cataract and vitreous surgery were also beneficial in the management of these patients. PMID- 3034223 TI - Leiomyoepithelioma of iris pigment epithelium. AB - A 4-year-old boy had a jet-black, nodular lesion growing in his right iris. Detailed clinical and histopathologic studies of this unusual tumor showed the lesion to be derived from the anterior iris pigment epithelium. The tumor cells appeared to be benign and exhibited characteristics of both pigment epithelium and smooth muscle. We have called this tumor "leiomyoepithelioma of iris pigment epithelium." PMID- 3034224 TI - The isolation of a bluetongue serotype new to Australia. PMID- 3034225 TI - Nephroblastoma in two dogs. PMID- 3034226 TI - Comparison of two commercial enzyme-linked immunosorbent assays and conventional methods for avian serology. AB - Sera tested for hemagglutination-inhibition (HI) activity against Newcastle disease virus (NDV) and infectious bronchitis virus (IBV) and virus-neutralizing (VN) activity against infectious bursal disease virus (IBDV) and viral arthritis (VA) virus were collected from a wide variety of accessions into the Diagnostic Services Laboratory, Poultry Disease Research Center, University of Georgia. The sera were then segregated according to HI or VN titer to NDV, IBV, IBDV, or VA virus and stored frozen at -20 C until tested by two commercial enzyme-linked immunosorbent assays (ELISAs). There was good correlation of mean Flockchek ELISA titers or EIA Systems sample-to-positive (S/P) ratios with specific HI or VN titers. Flockchek ELISA profile group 3 and EIA Systems mean S/P ratio of 1.12 corresponded to what were considered in our lab to be minimum protective titers for each antigen against virulent challenge in our area. PMID- 3034228 TI - Serological differentiation of avian infectious bronchitis field isolates using an enzyme immunoassay: presence of Dutch strains in West Germany. AB - Six field virus isolates were identified as the etiologic agent of avian infectious bronchitis (IB) in West Germany. A serological comparison was carried out using the M-41 strain from the Massachusetts serotype and the Dutch variant strains D-207, D-1466, D-3128, and D-3896 in a cross indirect immunoperoxidase test. Three isolates demonstrated similarity to the M-41 strain; the remainder showed a closer antigenic relationship to the Dutch variant strains. These results correspond to differences observed in a hemagglutination-inhibition test. The enzyme immunoassay is proposed as another possible method for differentiating IB virus strains. PMID- 3034227 TI - Pathological changes of tracheal mucosa in chickens infected with fowl pox virus. AB - Five-week-old chickens were inoculated with fowl pox (FP) virus and killed on various days through day 30 postinoculation (PI). The trachea was examined with a scanning electron microscope (SEM), a transmission electron microscope (TEM), and a light microscope (LM). From day 3 PI, small focal lesions of the mucosa were detected. On day 7 PI, upon formation of cytoplasmic inclusion bodies, epithelial cells proliferated profusely, enlarged, and formed clusters like papillomata. The disease proceeded to the gradual disruption of the lesions owing to the collapse of individual degenerating epithelial cells. Total desquamation of the lesions was observed. Ultrastructural examination revealed that the surface degenerating epithelial cells of the lesions ruptured and had virus particles inside. These changes were accompanied by severe inflammatory reaction. Thereafter, epithelial cells regenerated actively and the mucosa recovered by day 27 PI. PMID- 3034229 TI - Experimental infection of laying chickens with egg-drop syndrome 1976 virus. AB - Brown layer hens (BC and HC strains) and white layer hens (WL strain) orally infected with the H-162 strain of the egg-drop syndrome 1976 virus developed few clinical signs except for abnormal egg production. Depressed and/or aberrant-egg production was observed for 3 days or longer in 17 of 18 BC hens, 13 of 15 HC hens, and 10 of 17 WL hens. On the average, abnormal egg production began 8.8, 10.3, and 12.2 days after infection of the BC, HC, and WL hens, respectively. Egg production was depressed in the WL hens, but little depression was observed in the BC and HC hens. Aberrant-egg production was much less frequent in the WL hens than in the BC and HC hens. Aberrant eggs were shell-less, soft-shelled, thin shelled, and/or discolored. No eggs of abnormal internal quality or shape were observed. The virus spread from infected BC and WL hens to contact hens. PMID- 3034231 TI - Infectious bursal disease and spirochetosis in pullet chicks. AB - Infectious bursal disease and spirochetosis were diagnosed simultaneously in a flock of 100 six-week-old babcock pullet chicks. Larvae of the fowl tick Argas persicus were found on the bodies of the chicks. Thirty-nine of the chicks died 5 days after the onset of clinical signs. PMID- 3034230 TI - Infection of duck plague carriers with Pasteurella multocida and P. anatipestifer. AB - Mallards (Anas platyrhynchos platyrhynchos) and white pekin ducks (Anas platyrhynchos domesticus) were infected with duck plague virus and challenged with LD20's of Pasteurella multocida and P. anatipestifer. There was no difference between mortality rates of duck plague-infected ducks and controls, suggesting that these organisms do not act synergistically under the conditions of our experiments. There was a difference of about 500-fold between the LD20 of P. multocida for mallards and that for white pekin ducks, indicating that mallards are much more susceptible to avian cholera than white pekin ducks. PMID- 3034232 TI - A survey of enteric viruses of turkey poults. AB - Intestinal samples from 91 turkey flocks between 1 day and 5 weeks of age were examined for enteric viruses using electron microscopy and electropherotyping. These flocks originated from eight operations in six states. Individual flocks were sampled only once. At the time of sampling, 31 flocks were considered normal/healthy and 60 were considered to have enteric disease. The most frequently identified viruses from diseased flocks were astroviruses (78%) and rotavirus-like viruses (RVLVs) (67%). Far less frequent were rotaviruses (22%), atypical rotaviruses (12%), enteroviruses (5%), and reoviruses (2%). Only 10% of the samples from diseased flocks were negative, but 48% of the samples from normal/healthy flocks were negative. Astroviruses and RVLVs were far less frequent in normal/healthy flocks than in diseased flocks, but rotaviruses were identified slightly more often. No viruses were detected from flocks sampled within the first few days of life. Astrovirus infections seemed to occur at an earlier age than other virus infections. Seldom was only one type of virus identified. Astrovirus + RVLV was the most frequently identified combination in diseased flocks. PMID- 3034233 TI - [Synergistic and antagonistic hemolytic reactions of bacterial proteins]. PMID- 3034234 TI - Provisional assignment of the gene for uridine monophosphatase-2 (Umph-2) to mouse chromosome 11. AB - The segregation of the mouse gene for uridine monophosphatase-2 (Umph-2) was examined in 14 independent mouse-Syrian hamster hybrids and 10 hybrid subclones. Umph-2 cosegregated with the mouse galactokinase (Glk) gene in 23 of the 24 hybrids and showed at least four discordances with all other mouse marker isozymes examined. The observed synteny of Umph-2 and Glk, which has also been observed in humans, indicates that the mouse Umph-2 gene is on chromosome 11. PMID- 3034235 TI - Genetic variability in mitochondrial DNA in a regional population of the great tit (Parus major). AB - Genetic variation of mitochondrial DNA (mtDNA) in 18 great tits (Parus major) from three neighboring localities in Sweden was investigated with eight tetranucleotide restriction endonucleases. The 18 individuals could be separated into 13 different maternal lineages. The high number of female lineages present in this regional population contrasts with a low level of sequence divergence between the different mtDNA clones, with a mean of 0.19% sequence divergence between all individuals. There was no obvious spatial structuring of mtDNA clones among the three localities. The presence of a high number of different clones with a low degree of sequence divergence could be explained by the effects of a large long-term effective population size, with the mtDNA clones having diverged about 25,000-200,000 years ago. PMID- 3034236 TI - Structure and function of the small (30K) subunit of calcium-activated neutral protease (CANP). AB - Calcium-activated neutral protease (CANP), a typical intracellular protease, is composed of a catalytic 80K subunit (80K) and a 30K subunit (30K) of unknown function. The structure of rabbit CANP 30K was examined to clarify its role in the enzyme function. It has a clear two-domain structure composed of 266 amino acid residues. The N-terminal domain presumably determines the location of CANP, whereas the C-terminal domain is a calmodulin-like Ca2+-binding domain and regulates CANP activity. PMID- 3034237 TI - Protein products synthesized by a cloned viral protease gene. AB - A region of the poliovirus genome coding for a cysteine protease was expressed in E. coli. Following separation in isoelectric focussing gels, four new proteins were identified and were shown to be products of the viral cDNA. Although a majority of the protein products is aggregated, the system offers the first practical basis for production of large quantities of a viral protease. PMID- 3034238 TI - How does glucose initiate proteolysis of yeast fructose-1,6-bisphosphatase? AB - Glucose-induced proteolysis of fructose-1,6-bisphosphatase (FBPase) in yeast (Funayama, S. et al. (1980) Eur. J. Biochem. 109, 61-66) is preceded by rapid phosphorylation of the enzyme (Muller, D., and Holzer, H. (1981) Biochem. Biophys. Res. Commun. 103. 926-933; Mazon, M. J. et al. (1982) J. Biol. Chem. 257, 1128-1130). The postulated sequence of events is as follows. Glucose causes a decrease of the cytosolic pH from 7.0 to about 6.5 (Purwin, C. et al. (1986) J. Biol. Chem. 261, in press). The change in pH activates adenylate cyclase which is at pH 6.5 2-3 times more active than at pH 7.0 The concentration of cAMP increases 2-5 times (Purwin, C. et al. (1982) Biochem. Biophys. Res. Commun. 107, 1482-1489) and thereby activates a protein kinase which phosphorylates FBPase (Pohlig, G., and Holzer, H. (1985) J. Biol. Chem. 260, 13818 to 13823). The phosphorylated enzyme is either proteolyzed in the cytosol or taken up into the vacuoles, the compartment where unspecific proteolysis takes place. PMID- 3034239 TI - Studies on thiamine diphosphate kinase (EC 2.7.4.15) from brewer's yeast: purification and some properties. AB - Radiometric and fluorescent methods for detection of thiamine triphosphate have been used to show the presence of thiamine diphosphate kinase activity in brewer's yeast and to determine optimal conditions for its manifestation. A method of enzyme purification has been developed which involves glycerol-EDTA solution extraction, heat treatment, 2-fold ammonium sulphate fractionation, Sephadex G-200 gel filtration and ion-exchange chromatography on Sephadexes CP-C 50 and QAE-A-25. The protein has been purified 2000-fold in a 19% yield. The isoelectric and isoionic points, the amino acid composition and the molecular weight have been determined. Hydrophobic amino acids and those responsible for alpha-helix formation of the protein globule are predominant. The isoelectric point, as calculated by the amino acid composition and found by the maximum of the changes in fluorescence, is 5.8. The isoionic point value is identical with the isoelectric point. Upon gel filtration thiamine diphosphate kinase is eluted as two protein peaks with molecular weights of 162,000 +/- 8,000 and 81,000 +/- 4,000. After treatment with urea or sodium dodecyl sulphate, the protein dissociates into subunits with molecular weights of 12,500 and 14,000. The purified enzyme has some properties typical of a dissociating enzyme system. PMID- 3034240 TI - Alterations of surface charges of plasma lipoproteins in ischemic heart disease. AB - For a charged and an uncharged long chain spin probe the partition between the aqueous phase and the lipoproteins LDL, HDL2 and HDL3 was measured by use of ESR spectroscopy. The partition coefficients were compared for lipoproteins from normal donors and lipoproteins from patients with ischemic heart disease. The partition coefficients of the uncharged spin probe are not different. However, the charged spin probe has a significantly different partition for LDL and HDL3. This difference results from changes in the surface charge. Patients with ischemic heart disease have LDL which is more electropositively charged and HDL3 is more electronegatively charged compared to the corresponding lipoproteins of normal subjects. The surface charge of HDL2 is not changed. The results are discussed in the light of current concepts of the pathogenesis of atherosclerosis. PMID- 3034242 TI - Tyrosine phosphorylation of interleukin 2 receptor-related proteins in phytohemagglutinin-activated human lymphocytes. AB - Three classes of proteins (mol wts 70k, 64k and 45k) having the characteristics of interleukin 2 receptor were detected in phytohemagglutinin-activated human lymphocytes using two monoclonal antibodies which recognize distinct epitopes on the receptor. It was shown that at least portions of these proteins were phosphorylated on tyrosine by analyses for phosphotyrosine by immunoblotting and by immunoaffinity chromatography with antibodies to phosphotyrosine. In addition an iodinated phosphotyrosine derivative was identified in partial hydrolysates of these proteins iodinated in vitro. PMID- 3034243 TI - Spectral properties of cytochrome c' from Rhodopseudomonas capsulata B100 and its CO complex. AB - The spectral properties of cytochrome c' from photosynthetic bacterium Rhodopseudomonas capsulata (= Rhodobacter capsulatus) B100 and its CO complex are reported. The electronic absorption, MCD, and EPR spectra have been compared with those of the other cytochromes c' and horse heart cytochrome c. EPR and electronic spectral results for the ferric cytochrome c' suggest that the ground state of heme-iron(III) at neutral pH consists of a quantum mechanical admixture of an intermediate-spin and a high-spin state and that at pH 11.0 is in a high spin state. In the MCD spectrum of the CO-ferrous cytochrome c', the MCD intensity in the Soret band region was much higher than that of CO complexes of hemoproteins with a protoheme. The differences in a stereochemistry of the sixth coordination position is discussed. PMID- 3034241 TI - Inhibition of human neutrophil collagenase by gold(I) salts used in chrysotherapy. AB - Six gold(I) salts, some of which are used as drugs in chrysotherapy, are shown to be inhibitors of two forms of human neutrophil collagenase. The IC50 values vary over six orders of magnitude, the lowest being 3.5 nM for Myocrisin. Thus, inhibition is greatly affected by the identity of the ligands to the gold(I) atom. The inhibition of collagenase by these gold(I) salts may be a partial basis for their antiarthritic action. PMID- 3034244 TI - Effect of a high carbohydrate diet on cardiac alpha-1 and beta adrenoceptors. AB - Short-term (2 weeks) administration of a high-fructose diet to euthyroid Sprague Dawley rats results in a significant (18%) increase of cardiac beta-adrenoceptors without any change in their affinities or their distribution between the plasma membrane and a vesicular fraction. A much smaller increase occurs in the number of alpha 1-adrenoceptors (8% higher than in rats fed a regular diet). The high carbohydrate diet induced a 39% increase in beta-adrenoceptor numbers/heart in hypothyroid animals and a 19% increase in the total number of alpha 1 adrenoceptors. These results strongly suggest that changes in cardiac performance after dietary manipulations may be mediated, in part, through enhancement of adrenergic pathways. PMID- 3034245 TI - Two dimensional NMR studies on the solution structure of d-CTCGAGCTCGAG. AB - Two dimensional (2D) FT-NMR investigations have been carried out on the self complementary dodecanucleotide d-CTCGAGCTCGAG, which has cleavage sites for the restriction enzyme Xho I (between C and T). The central TCG portion is also known to show a preference for DNAase activity. Complete resonance assignments have been obtained for the non-exchangeable sugar and base protons of the oligonucleotide. Information regarding sugar geometries, glycosidic torsion angles and other structural parameters has been obtained from the relative intensities of the cross peaks in the COSY and NOESY spectra. The results indicate that deoxyribose rings of C1 and C7 adopt a conformation different from the remaining sugars in the double helical oligonucleotide. The central TCG portion also exhibits variations in the backbone structure. The base stacking in the double helix shows interesting sequence dependent effects suggesting that the sequence effects are not localised to nearest neighbours but extended over longer stretches. PMID- 3034246 TI - Identification of a novel inositol bisphosphate isomer formed in chemoattractant stimulated human polymorphonuclear leukocytes. AB - Analysis of inositol phosphate formation in chemoattractant-stimulated human polymorphonuclear leukocytes demonstrated the production of inositol 1,4,5 trisphosphate, inositol 1,3,4-trisphosphate, inositol 1,3,4,5-tetrakisphosphate, inositol 1,4-bisphosphate and another inositol bisphosphate isomer not detected in unstimulated cells. Studies in cell sonicates provided evidence that the previously unidentified inositol bisphosphate isomer is produced via the degradation of inositol 1,3,4-trisphosphate. This unidentified inositol bisphosphate peak was purified by high pressure liquid chromatography, and base hydrolyzed to form a mixture of inositol monophosphate isomers. Based on these studies, the unidentified peak was identified as inositol 3,4-bisphosphate. Identification of this isomer defines a new metabolic product derived from the initial inositol 1,4,5-trisphosphate formation, and also suggests another substrate for the inositol 1-phosphatase. PMID- 3034247 TI - Stabilized ubisemiquinone in reconstituted succinate ubiquinone reductase. AB - QP-S, a ubiquinone (Q) protein, accepts electrons from succinate through succinate dehydrogenase (SDH). A new method has produced a preparation of QP-S which has a different amino acid composition and SDS gel electrophoretic pattern from that of the old preparation (Biochemistry 19, 3579-3585 (1980)). The new preparation contains less than 1 nmol heme/mg protein; the activity of the preparation was not proportional to its heme content. A thenoyltrifluoroacetone sensitive free radical signal was detected by EPR spectroscopy in succinate-Q reductase reconstituted from this QP-S and SDH; the characteristics of this species identify it as ubisemiquinone. At pH 7.4, the Em of the two electron step was about 70 mV with E1 = 5 mV and E2 = 125 mV. The properties of the radical differed slightly from those of "Qs" radical in more intact preparations (e.g. submitochondrial particles). The present is the simplest system in which such a succinate reducible ubisemiquinone free radical has been demonstrated. PMID- 3034249 TI - Divergence of ANF analogs in smooth muscle cell cGMP response and aorta vasorelaxation: evidence for receptor subtypes. AB - ANF analog potencies in stimulating smooth muscle cell cGMP were compared with the ability to relax histamine-constricted rabbit aorta in vitro. ANF[1-28], [5 28], [5-27] and Lys-11[5-28] elevated cGMP and were potent vasorelaxants. ANF[7 23] and Lys-11[7-23] were potent cGMP stimulators but 1000-fold weaker relaxants. Tyr-8[5-27] did not stimulate cGMP synthesis or antagonize the response of the other peptides, yet was a potent vasorelaxant. Crosslinking with 125I-ANF identified bands at 150 and 65 KD by SDS-PAGE. ANF[1-28], Lys-11[7-23] and Tyr 8[5-27] blocked crosslinking at low concentration despite disparate activities. These data support the existence of ANF receptor subtypes and suggest that cGMP elevation alone is not sufficient to promote atrial peptide-induced vasorelaxation. PMID- 3034248 TI - A novel mechanism for utilization of extracellular AMP in Vibrio parahaemolyticus. AB - Vibrio parahaemolyticus could grow with AMP, ADP or ATP as the sole source of carbon. In the presence of Cl-, a membrane-bound Cl(-)-dependent 5'-nucleotidase seemed to hydrolyze the nucleotides extracellularly, and then the cells took up the resulting adenosine. In the absence of Cl-, although no significant dephosphorylation of the nucleotides occurred, the cells could still grow with AMP, but not with ADP or ATP. Moreover, in the presence of Cl-, Zn2+ inhibited the 5'-nucleotidase, and inhibited growth of the cells with ADP or ATP, but not with AMP, as the carbon source. V. parahaemolyticus was unable to grow with adenine or ribose 5-phosphate. These results suggested that the cells might have an AMP transport system. In fact, Na+ uptake was observed on addition of AMP to a cell suspension in the absence of Cl-, indicating Na+-AMP cotransport. PMID- 3034251 TI - Neomycin inhibits agonist-stimulated polyphosphoinositide metabolism and responses in human platelets. AB - The effect of neomycin on agonist-induced changes in polyphosphoinositide metabolism was studied in human platelets. Neomycin induced no changes in the 32P labeled phospholipids of unstimulated cells. Between 1 and 5 mM of neomycin the initial thrombin-induced decrease in [32P]phosphatidylinositol-4,5-bis-phosphate and production of [32P]phosphatidic acid were gradually inhibited. In contrast, the production of [32P]phosphatidylinositol-4-phosphate was increased. The thrombin-induced decrease in mass of phosphatidylinositol was completely inhibited at 5 mM of neomycin. Collagen- and PAF acether-induced changes in platelet phosphoinositide metabolism as well as aggregation and dense granule secretion induced by all three agonists were inhibited by neomycin. The results indicate that neomycin inhibit platelet responses by selective interference with the interconversions and hydrolysis of polyphosphoinositides upon thrombin stimulation. PMID- 3034250 TI - Histamine-induced phosphoinositide metabolism in cultured human umbilical vein endothelial cells. Association with thromboxane and prostacyclin release. AB - Histamine stimulation of cultured human umbilical vein endothelial cells induced dose- and time-dependent increases in glycerophosphoinositol (GroPIns), inositol 1-phosphate (InsP), inositolbisphosphate (InsP2) and inositoltrisphosphate (InsP3) in addition to release of thromboxane A2 and prostacyclin. Increases in InsP2 and InsP3 were immediate while increases in GroPIns and InsP occurred only after 1 min. Thromboxane A2 and prostacyclin release paralleled GroPIns and InsP production. The data indicate that, in endothelial cells, histamine evokes early hydrolysis of polyphosphoinositides, and that subsequent mobilization of arachidonic acid for thromboxane and prostacyclin synthesis involves both deacylation and phosphodiesteratic cleavage of phosphatidylinositol. PMID- 3034252 TI - Genetic heterogeneity of steroid sulfatase deficiency revealed with cDNA for human steroid sulfatase. AB - Three cDNA clones with inserts of 1.2-1.6 kb that reacted both with antibodies and oligonucleotides specific for steroid sulfatase were isolated from a human placental library in lambda gt11. The 5'-end of one of the inserts, STS-3, was sequenced and colinearity with the amino acid sequence of 3 peptides of steroid sulfatase encompassing 64 amino acids was demonstrated. STS-3 hybridized with 2.5, 4.6 and 6.3 kb species in poly(A)+RNA and with 2.5, 4 and 9 kb fragments of EcoRI digested human DNA. The frequency of the EcoRI fragments in DNA from females was approximately twice that in DNA from males. DNA from two patients with steroid sulfatase deficiency and X-linked ichthyosis did not hybridize with STS-3. DNA from a third patient showed a normal hybridization pattern. It is concluded that steroid sulfatase deficiency is a genetically heterogenous disorder. PMID- 3034253 TI - Enriched autonomously replicating sequences in a nuclear matrix-DNA complex isolated from synchronized HeLa cells. AB - Nucleoids isolated from either synchronized or exponentially growing HeLa cells were digested with restriction enzymes to separate a nuclear matrix-bound DNA component from the rest. Partial libraries were constructed by inserting DNA fragments from both components into a yeast-bacteria plasmid vector. A random sample from these libraries was tested for ARS activity by a standard yeast transformation assay. We found that synchronization for DNA replication results in an enrichment for autonomously replicating sequences in the library constructed with the DNA component bound to the nuclear matrix. PMID- 3034254 TI - Translocation of diacylglycerol kinase in response to chemotactic peptide and phorbol ester in neutrophils. AB - When neutrophils were stimulated by the chemotactic peptide, fMLP, a rapid, transient increase in the activity of diacylglycerol(DG) kinase in the membrane fraction was detected. DG kinase in cytosol, on the contrary, showed a transient decrease. The total activity in homogenates was not affected. Tetradecanoylphorbol acetate(TPA) and 1-oleoyl-2-acetylglycerol(OAG) also caused an increase in DG kinase activity in the membrane fraction. Km value of DG kinase in membranes was not changed by the treatment of fMLP or TPA, though Vmax was increased. Considering these results, DG kinase may translocate from cytosol to membranes on stimulation by fMLP, TPA or OAG in neutrophils. The translocation may play important roles in regulation of protein kinase C activity, since DG kinase competes with protein kinase C for DG, which is formed by receptor activation. PMID- 3034255 TI - Sulfinyl radical formation from the reaction of cysteine and glutathione thiyl radicals with molecular oxygen. AB - Using Electron Spin Resonance spectroscopy at low temperatures, we find that thiyl radicals resulting from irradiation of frozen aqueous solutions of a variety of thiols, including cysteine, glutathione, and penicillamine react with oxygen to form sulfinyl (RSO.) radicals. The identity of the cysteine sulfinyl radical has been confirmed by the use of molecular oxygen isotopically labeled with 17O. Previous workers have suggested the reaction of thiyl radicals and molecular oxygen resulted in the formation of the potentially damaging thiol peroxyl radical, RSOO.; our work shows no evidence for this species. The sulfinyl radicals are suggested to result from a direct reaction between thiyl radicals and molecular oxygen. This reaction results in the cleavage of the dioxygen bond. PMID- 3034256 TI - Binding characteristics of atrial natriuretic factor and the production of cyclic GMP in kidneys of Dahl salt-sensitive and salt-resistant rats. AB - The binding of atrial natriuretic factor (ANF) was studied in kidney membranes of inbred salt-sensitive (S) and inbred salt-resistant (R) rats on high or low salt diet. Important differences between strains were seen in the rate of dissociation of ANF from its renal receptor(s) and this was dependent on salt (NaCl) intake. On low salt diet ANF dissociation rates were similar between strains. R rats responded to high salt diet with a decrease in the rate of ANF dissociation from its renal receptor, but ANF dissociation in S rats was not altered by dietary salt. Receptor density was similar between strains. Basal cGMP production was slightly higher for renal membranes of S rats, but ANF stimulation of cGMP production was similar between strains and was not influenced by salt intake in either strain. Since strain-related salt-induced changes in ANF-receptor binding kinetics were not reflected in any strain-related salt-induced changes in ANF stimulated cGMP production, it is tentatively concluded that the ANF receptor likely to be different between S and R strains is the ANF receptor not linked to cGMP production. PMID- 3034257 TI - Stimulation of procollagenase synthesis parallels increases in cellular procollagenase mRNA in human articular chondrocytes exposed to recombinant interleukin 1 beta or phorbol ester. AB - Interleukin 1, a product predominantly of monocytes, increases the synthesis and release of procollagenase and prostaglandin E2 by mesenchymal target cells such as synovial fibroblasts and articular chondrocytes, an effect mimicked by some phorbol esters. In order to determine the mechanisms underlying these responses primary cultures of human articular chondrocytes were preincubated with recombinant human interleukin 1 beta or the phorbol ester, phorbol 12-myristate 13-acetate, in the presence or absence of the cyclooxygenase inhibitor, indomethacin. Interleukin 1 beta or phorbol ester increased the levels of procollagenase (assayed after trypsin activation) and the labeling of several medium proteins by cells incubated with [35S]methionine, independent of prostaglandin synthesis. The labeling of a 55 kD protein immunocomplexed with antibodies to procollagenase was also increased. The increased synthesis of procollagenase was paralleled by increased cellular levels of procollagenase mRNA, determined with a cDNA probe coding for human procollagenase. Thus the increased synthesis of procollagenase in response to the inflammatory mediator, interleukin 1, is controlled at a pretranslational level, possibly at the level of transcription. PMID- 3034258 TI - Saturated and trans-unsaturated fatty acids elicit high levels of superoxide generation in intact and cell-free preparations of neutrophils. AB - Saturated and trans-unsaturated fatty acids, such as laurate and elaidate, elicited O2- generation in intact porcine and human neutrophils and also in a cell-free preparation of porcine neutrophils. The activities thus induced were comparable to those induced by cis-unsaturated fatty acids. However, the activation by saturated or trans-unsaturated fatty acids was depressed almost completely in the presence of Ca2+ at around 1 mM, which is usually contained in the media for phagocytes. In contrast, the activation by cis-unsaturated fatty acids such as arachidonate was scarcely affected by Ca2+. These findings appear to demand reevaluation of the effects of long chain fatty acids on the respiratory burst system in phagocytes. PMID- 3034260 TI - The anesthetic nitrous oxide affects dioxygen utilization by bovine heart and bean seed mitochondrial particles. AB - Nitrous oxide affects dioxygen utilization by both bean seed and bovine heart submitochondrial particles when either succinate or reduced cytochrome c are used as substrates. Bovine heart particles exhibit reversible, dose-dependent partial inhibition of respiratory activity when exposed to N2O. Bean seed particle respiration is stimulated by low levels of N2O, but higher concentrations are inhibitory. These findings can be explained in terms of one locus of anesthetic action: cytochrome c oxidase, the terminal component of the mitochondrial respiratory chain. Alterations in respiration rates are expected to make important contributions to anesthesia in animals and to control of germination in plants. PMID- 3034259 TI - Down-regulation of protein kinase C in rat glioma C6 cells: effects on the beta adrenergic receptor-coupled adenylate cyclase. AB - Continuous exposure of rat glioma C6 cells to 12-O-tetradecanoylphorbol-13 acetate (TPA) resulted in a time and dose dependent loss of [3H]phorbol dibutyrate binding sites and protein kinase C activity. Thus, by 24 h, the cells were essentially depleted of protein kinase C activity. In agreement with previous studies, TPA treatment caused a reduction in isoproterenol-stimulated adenylate cyclase activity and a sequestration of beta-adrenergic receptors. Cells were treated with TPA for 24-48 h to completely down-regulate protein kinase C and then exposed to isoproterenol. Agonist-mediated desensitization of adenylate cyclase and sequestration of beta-adrenergic receptors occurred at similar rates in control and TPA-treated cells. In addition, agonist-mediated down-regulation of beta-adrenergic receptors was not impaired by the absence of protein kinase C activity. Although both agonists and phorbol esters cause desensitization of the beta-adrenergic receptor-coupled adenylate cyclase, agonist-mediated events can occur independently of protein kinase C. PMID- 3034261 TI - Calcitriol increases Ca2+-ATPase activity. AB - Treatment with calcitriol of isolated cartilage cells derived from epiphyseal growth plates of rachitic chicks results in reduced intracellular calcium concentrations. The reduction in calcium was found to correlate with increased activity of Ca2+-ATPase. The activities of Na+-K+-ATPase and of Mg2+-ATPase did not change in response to the treatment with calcitriol. It is suggested that calcitriol regulates intracellular calcium by modulating the activity of the Ca2+ pumping ATPase. PMID- 3034262 TI - Defective adaptation to a low phosphate environment by cultured renal tubular cells from X-linked hypophosphatemic (Hyp) mice. AB - Effects of parathyroid hormone (PTH), low phosphate environment, and 12-O tetradecanoyl phorbol-13-acetate (TPA) on the phosphate reabsorption by the renal tubular cells from mutant hemizygous hypophosphatemic (Hyp/Y) mice and their littermates (+/Y) were studied using a phosphate accumulation system which had been developed recently. This system mimics phosphate transport at the renal tubules. When cultured in a normal phosphate medium, the characteristics of the phosphate accumulation by Hyp cells was almost identical with that by normal cells; a PTH-induced inhibition and a TPA-induced stimulation of phosphate accumulation. However, when preincubated in a low phosphate medium, the accumulation of phosphate by normal cells increased significantly, while that by Hyp cells did not. These results indicate that the adaptation to the low phosphate environment is defective in Hyp cells and it may be one of the cause of renal phosphate leakage in the Hyp mouse. PMID- 3034263 TI - Electron spin resonance (ESR) studies of skeletal protein interactions in human erythrocyte membranes exposed to polyanions and in membranes prepared from inositol hexaphosphate (IHP)-incorporated low-affinity erythrocytes. AB - Previous biophysical investigations, including those from our laboratories, have reported that polyphosphates weaken RBC membrane skeletal protein-protein interactions and decrease hemoglobin affinity for oxygen. We have additionally demonstrated that low-affinity intact RBC's may be produced by inositol hexaphosphate (IHP) incorporation via an osmotic pulse method. In the present electron spin resonance (ESR) study, IHP was shown to cause a concentration dependent increase in the segmental motion of ghost membrane skeletal proteins, but no alterations in spin-labeled terminal sialic acid. Pyrophosphate and inositol hexasulfate were significantly less effective in altering the physical state of skeletal proteins than was IHP. Additional ESR studies of both the interaction of IHP with membrane skeletal proteins in the presence of hemoglobin and of membranes obtained from osmotic pulse-treated intact cells were performed. The results of all these studies are discussed in terms of previous biophysical investigations of the effects of polyphosphates on membranes and of possible molecular events that occur during the osmotic pulse procedure. PMID- 3034264 TI - The interaction between nuclear topoisomerase II activity from human leukemia cells, exogenous DNA, and 4'-(9-acridinylamino)methanesulfon-m-anisidide (m-AMSA) or 4-(4,6-O-ethylidene-beta-D-glucopyranoside) (VP-16) indicates the sensitivity of the cells to the drugs. AB - The presumptive intracellular target of the anti-leukemia agents 4'-(9 acridinylamino)methanesulfon-m-anisidide (m-AMSA) and 4-(4,6-O-ethylidene-beta-D glucopyranoside) (VP-16) is the enzyme topoisomerase II. We found that 350 mM NaCl extracts of nuclei from HL-60 and HL-60/AMSA, an m-AMSA resistant HL-60 subline, contained equivalent topoisomerase II activity. However, the ability of m-AMSA to stimulate cleavage of exogenous DNA and to stimulate crosslinking of exogenous DNA with protein, processes which are topoisomerase II-mediated, was greatly reduced in the HL-60/AMSA extracts compared to the HL-60 extracts. HL-60 and HL-60/AMSA were almost equally sensitive to the cytotoxic effects of VP-16 and differences in VP-16-stimulated, topoisomerase II-mediated exogenous DNA cleavage and protein crosslinking between HL-60 and HL-60/AMSA extracts were much less than the differences in m-AMSA-stimulated exogenous DNA cleavage and protein crosslinking. Thus, the interaction between topoisomerase II activity, exogenous DNA, and m-AMSA or VP-16 indicated the susceptibility HL-60 and HL-60/ AMSA to the cytotoxic effects of the drugs. A similar correlation may exist in explanted leukemia cells from patients with acute myelogenous leukemia. PMID- 3034266 TI - Species differences between rat and rabbit renal Na+/H+ exchangers. AB - Amiloride, 5-ethylisopropylamiloride (EIPA), and 5-ethylisopropyl-6-bromo amiloride (Br-EIPA) inhibit rabbit and rat renal Na+/H+ exchangers with comparable potency. Irreversible inhibitions by Br-EIPA after irradiation of rat and rabbit renal brush-border vesicles are also similar. In contrast, irreversible inhibition by N,N'-dicyclohexylcarbodiimide (DCCD) of the rabbit renal antiporter requires higher DCCD concentrations as compared to the rat. Rabbit renal brush-border membranes show highest [14C]DCCD incorporation at MW 80,000, 51,000 and 36,000 and lack the amiloride-protectable MW 65,000 protein previously identified as Na+/H+ exchanger in rat kidney cortex. The data indicate species differences with respect to renal Na+/H+ exchangers. PMID- 3034267 TI - Stimulation of leucine transport by a phorbol ester through activation of protein kinase C and Na+/H+ exchanger. AB - A synthetic diacylglycerol, 1-oleoyl-2-acetyl-sn-glycerol (OAG), as well as 12-O tetradecanoylphorbol-13-acetate (TPA) has been found to elevate the cytoplasmic pH and increase leucine uptake dose-dependently, when added to quiescent cultures of Chang liver cell. Addition of either a protein kinase C inhibitor, 1-(5 isoquinolinylsulfonyl)-2-methylpiperazine (H-7), or an Na+/H+ antiporter inhibitor, ethylisopropylamiloride (EIPA), abolished completely or incompletely the TPA-stimulated leucine uptake and TPA-induced cytoplasmic alkalinization. Therefore the stimulation of leucine uptake by OAG and TPA is proposed to be elicited at least partly through activation of Na+/H+ antiporter. We suggest that activation of protein kinase C by the phorbol ester is responsible for the stimulation of Na+/H+ exchange system and also leucine uptake in the cell. PMID- 3034265 TI - Lack of correlation between calcium mobilization and respiratory burst activation induced by chemotactic factors in rabbit polymorphonuclear leukocytes. AB - Low concentrations of FMLP, partially purified rabbit C5a, leukotriene B4 and platelet activating factor induced a rapid rise of intracellular free Ca2+ in rabbit polymorphonuclear leukocytes. However, the four factors differed markedly in their ability to activate the respiratory burst. The peptides FMLP and C5a induced a single, strong chemiluminescence response whereas the lipids leukotriene B4 and platelet activating factor induced a markedly less intense response with a two-peak profile. Respiratory burst activation by the peptides was dependent on extracellular Ca2+ whereas the lipids required both Mg2+ and Ca2+. The results indicate that mobilization of intracellular Ca2+ is insufficient by itself to induce respiratory burst activation and that the intracellular pathways leading to activation differ depending on the nature of the stimulus. PMID- 3034268 TI - Angiotensin converting enzyme predominates in the inner cortex and medulla of the rat kidney. AB - Regional distribution of angiotensin converting enzyme(ACE) in the rat kidney was studied. The ACE activities in the inner cortex and outer medulla were about 10 and 5 times those in the outer cortex, respectively. The activity in the inner medulla or papilla was much the same as that in the outer cortex. Immunofluorescence was greatest in the proximal tubules in the inner cortex, while the outer medulla and the inner medulla or papilla showed a weak fluorescence. The brush border membranes isolated from the inner cortex also possessed about 10 times the ACE activity seen in the outer cortex. The results indicate that the major source of renal ACE is not the proximal convoluted tubules in the outer cortex, but rather the brush border membranes of proximal tubules in the inner cortex. The contribution of ACE in the inner cortex would therefore be predominant. PMID- 3034270 TI - Biochemical properties associated with the immortalizing domain of the large T protein of polyoma virus. AB - A fragment of polyoma virus DNA lacking the carboxy-terminal part of the large T antigen coding region is sufficient to express the functions of the entire gene in cell transformation. Two cell lines expressing this truncated DNA were studied, one of them producing a large T-related protein of the expected size (37 kDa) and the other one, a shorter product (34 kDa). Both proteins were phosphorylated and localized in the nucleus, but devoid of ATPase and nucleotide binding activities. As the complete protein, the larger product, but not the shorter variant, exhibited sequence-specific DNA binding properties. These results indicate that ATPase and nucleotide-binding activities are not required for immortalization, and suggest that recognition of specific DNA sequences may be dispensable. PMID- 3034269 TI - A proton magnetic resonance study of the effects of polyamine and divalent metal ions on diadenosine 5',5'''-P1,P4-tetraphosphate base stacking. AB - Complexation of putrescine, spermidine, spermine, and Mg2+ with diadenosine 5',5'''-P1,P4-tetraphosphate induces an upfield shift in the signals for the H-2 and H-8 protons. The upfield shifts in H-2 indicate that cation complexation enhances intramolecular adenine stacking interactions. The resonances for H-2 and H-8 of neutral analogs of 5',5'-dinucleotides appear farther upfield relative to the appropriate monomeric models than those for the corresponding dinucleotide; reduction of intra-chain phosphate repulsion is the origin of cation induced enhancement of diadenosine 5H,5'''-P1,P4-tetraphosphate base stacking. PMID- 3034271 TI - Human creatine kinase: isolation and sequence analysis of cDNA clones for the B subunit, development of subunit specific probes and determination of gene copy number. AB - cDNA clones for human B creatine kinase were isolated from human brain and placenta libraries. The entire coding and 3' untranslated regions, as well as 23 bp of the 5' untranslated region were sequenced. Complete sequence identity was found among the clones, with the exception of an area of heterogeneity among the 3' untranslated region of the brain and placenta clones. A 77.7% nucleotide sequence identity was found between the coding region of human B creatine kinase and our previously reported human M creatine kinase. In contrast, no homology was found in the 3' untranslated regions. Probes were constructed from the nonconserved 3' untranslated regions of human M and B creatine kinase and were shown to be highly specific. Southern transfers of total genomic DNA derived from human placenta and digested to completion with several restriction enzymes were probed with the MCK and BCK specific probes producing single hybridization bands. These results suggest that creatine kinase M and B are single copy genes in the human genome. PMID- 3034272 TI - Recombinant human granulocyte colony-stimulating factor enhances superoxide release in human granulocytes stimulated by the chemotactic peptide. AB - Recombinant human granulocyte colony-stimulating factor (G-CSF) by itself was not an effective stimulus for inducing the release of superoxide (O-2) in human granulocytes. However, G-CSF was able to prime human granulocytes, and enhanced O 2 release stimulated by the chemotactic peptide, N-formyl-methionyl-leucyl phenylalanine (FMLP). The preincubation with G-CSF for 5-10 min at 37 degrees C was sufficient for priming the cells. The optimal enhancing effect was obtained at 25 ng/ml of G-CSF. The enhancement of O-2 release by G-CSF was observed over the complete range of effective concentrations of FMLP (10(-8)-10(-6) M). These findings indicate that G-CSF is a potent activator of mature granulocyte functions. PMID- 3034274 TI - Purification of the catalytic subunit of protein phosphatase-1 from Drosophila melanogaster. AB - The catalytic subunit of phosphatase-1 has been purified from Drosophila melanogaster by precipitation with (NH4)2SO4 and ethanol, by affinity chromatography on heparin-Sepharose and by fast protein liquid chromatography on Mono Q beads. The preparation is homogeneous as tested by SDS gel electrophoresis and has a molecular mass of 33,000. The phosphatase specifically dephosphorylates the beta subunit of phosphorylase kinase. Its phosphorylase phosphatase activity is inhibited by inhibitor-1, inhibitor-2, protamine and histone H2B while is stimulated by histone H1. PMID- 3034275 TI - The bioflavonoid quercetin inhibits neutrophil degranulation, superoxide production, and the phosphorylation of specific neutrophil proteins. AB - Quercetin, a C-kinase antagonist, inhibits neutrophil degranulation and superoxide production induced by f-met-leu-phe, solid phase IgG, zymosan treated serum and a phorbol ester (PMA). Quercetin is more effective in inhibiting degranulation (IC50 = 20 uM) than superoxide production (IC50 = 80 microM). Neutrophil activation by PMA is accompanied by the phosphorylation of neutrophil proteins of 205, 170, 130, 91, 77, 67, 56, 47, 39, 34, 27, and 20 kilodaltons; quercetin also inhibits the phosphorylation of these proteins. Dose-response studies indicated that phosphorylation of the 67 kilodalton protein was particularly sensitive to inhibition by quercetin at concentrations that also inhibit neutrophil degranulation and superoxide production. These results suggest that phosphorylation of the 67 kilodalton protein may be an important intracellular reaction associated with neutrophil activation. PMID- 3034273 TI - A requirement for cholesterol for phorbol ester-induced adhesion of a human monocyte-like cell line. AB - The human monocyte/macrophage-like cell line U937, which is a cholesterol auxotroph, is nonadherent. However, it becomes adherent after treatment with phorbol 12-myristate 13-acetate (phorbol ester). We investigated the effects of cellular cholesterol depletion and repletion on the effectiveness of phorbol ester to induce adhesion to substratum. Almost 70% of cellular cholesterol is depleted by incubation of the cells for 24 hrs in the growth medium in which delipidated fetal calf serum is substituted for fetal calf serum without affecting viability or the rate of growth. The use of delipidated fetal calf serum inhibited phorbol ester-induced adhesion by 40%. If the cells were preincubated in the medium containing delipidated fetal calf serum 6 hrs prior to addition of phorbol ester, adhesion was inhibited by 90%. Addition of cholesterol to the medium containing delipidated fetal calf serum, which replenishes cellular cholesterol, restored the ability of phorbol ester to induce adhesion to levels seen in cells cultured in the medium containing fetal calf serum. Epicholesterol was not as effective as cholesterol in supporting adhesion. Cholesterol depletion did not inhibit phorbol ester stimulation of superoxide anion production. These observations indicate a function for cholesterol in phorbol ester-induced adhesion that is independent of phorbol ester-induced superoxide anion production. It is proposed that cholesterol is required for synthesis and/or proper orientation and distribution, in the plasma membrane, of macromolecule(s) that mediate phorbol ester-induced adhesion. PMID- 3034276 TI - Concanavalin A prevents phorbol-mediated redistribution of protein kinase C and beta-adrenergic receptors in rat glioma C6 cells. AB - Exposure of rat glioma C6 cells to the phorbol ester 12-O-tetradecanoylphorbol 13 acetate (TPA) caused an activation of protein kinase C wherein the enzyme rapidly became membrane-bound (T 1/2 of 15 min). This translocation of protein kinase C from cytosol to membrane was followed by a sequestration of cell surface beta adrenergic receptors and a loss of isoproterenol-stimulated adenylate cyclase activity. We had reported previously that prior exposure of rat glioma cells to concanavalin A prevents the TPA-mediated sequestration of receptors and desensitization of adenylate cyclase (Kassis et al., 1985). We now show that the concanavalin A treatment also prevents the translocation and activation of protein kinase C. These results are further evidence that in the TPA-treated cells, sequestration of beta-adrenergic receptors is mediated by membrane-bound protein kinase C. PMID- 3034277 TI - Effects of the protein kinase C inhibitor H-7 and calmodulin antagonist W-7 on superoxide production in growing and resting human histiocytic leukemia cells (U937). AB - Serum, phorbol 12,13-didecanoate (PDD) and 1-oleoyl-2-acetoy-sn-glycerol (OAG) stimulated O2- release in human histiocytic leukemia U937 cells. The kinetics of O2- release caused by PDD but not by serum or OAG in growing cells differed from those in resting cells. Both the protein kinase C inhibitor 1-(5 isoquinolinylsulfonyl) 2-methylpiperidine (H-7) and calmodulin antagonist N-(6 aminohexyl)-5-chloro-1-naphthalenesulfonamide (W-7) reduced the superoxide generation induced by these stimuli. H-7 inhibited the O2- release either from growing or resting cells but the effect of W-7 varied according to the growth phase. From these results, it is suggested that activation of protein kinase C and calmodulin-dependent process has an important role in O2(-)-release induced by serum, OAG and PDD, and that the mechanism for PDD-induced O2(-)-release is different in growing and resting cells. PMID- 3034279 TI - Transport of fatty acid is obligatory coupled with H+ entry in spheroplasts of Escherichia coli K12. AB - Transport of palmitate by spheroplasts of Escherichia coli K12 was studied. [14C]Palmitate was accumulated in spheroplasts approximately 1700-fold over the extracellular concentration of unbound [14C]palmitate. Uptake of [14C]palmitate was inhibited to 13% by addition of H+ uncoupler carbonyl cyanide-m chlorophenylhydrazone (CCCP). Spheroplasts exhibited the uptake of 9 aminoacridine depending on the addition of palmitate to the incubation mixture. The rate of [14C]palmitate uptake by the spheroplasts pre-equilibrated in a buffer at pH 7.5 or 8.0 significantly increased in comparison with the spheroplasts pre-equilibrated in a buffer at pH 7.0 when the spheroplasts were incubated at an external pH of 7.0. PMID- 3034278 TI - The human apoB-100 gene: apoB-100 is encoded by a single copy gene in the human genome. AB - Northern blot analysis of human liver and intestine mRNA revealed two separate apoB mRNA of 14.1 and 7.5 kb in the intestine, and a single 14.1 kb apoB mRNA in the liver. cDNA probes which encode for the 5', middle, and 3' regions of the human apoB-100 mRNA have been utilized to evaluate the number of apoB genes present in the human genome by Southern blot hybridization analysis. Comparison of restriction enzyme digestions of high molecular weight leukocyte DNA and a known apoB genomic clone with cDNA probes for the 5', middle and 3' regions of the apoB-100 mRNA were consistent with only a single apoB gene per human haploid genome. Further analysis with synthetic oligonucleotides definitively established that there is a single apoB-100 gene. The two mRNA species observed in the intestine must be derived from a single nuclear apoB RNA transcript. PMID- 3034280 TI - Detection of aspartate kinase in rat liver and its activation by Ca++ and calmodulin. AB - The enzyme aspartate kinase (EC 2.7.2.4) has been detected in rat liver (animal tissue) for the first time. This enzyme, like the aspartate kinase from bacteria and plants is inhibited by lysine and threonine. Further, the activity of the enzyme is stimulated over two fold by Ca++ and calmodulin and inhibited by EGTA, a Ca++ chelator and trifluoperazine, an anti-calmodulin compound. PMID- 3034282 TI - Molecular neurobiology. PMID- 3034281 TI - Activation of rat parotid low Km cyclic AMP phosphodiesterase by isoproterenol. AB - Approximately 94% of rat parotid cyclic AMP phosphodiesterase activity measured at a substrate concentration of 0.1 microM cyclic AMP was found in the 100,000 X g supernatant while the remaining enzyme activity was in the particulate fraction. Incubation of parotid slices with 10 microM isoproterenol resulted in approximately 40% activation of the cyclic AMP phosphodiesterase activity of the 100,000 X g supernatant. The enzyme activity in the particulate fraction was unaffected. The activation resulted from an increase in the value of the Vmax while the apparent Km (0.51 microM) was unaffected. The concentration of isoproterenol required to give half-maximal activation was 0.34 microM. The activation was rapid, became significant after 2 min and reached maximum after 30 min incubation of the parotid slices with isoproterenol. The activation of the enzyme activity by isoproterenol could be blocked by propanolol but was unaffected by cycloheximide. Dibutyryl-cyclic AMP was also effective while phenylephrine and carbamylcholine were ineffective in increasing the activity of the enzyme. PMID- 3034283 TI - Alpha and beta adrenergic and muscarinic cholinergic receptor structure. AB - Purification and characterization of the neurotransmitter receptors of the autonomic nervous system have revealed considerable structural and functional homology between these pharmacologically distinct classes of information transduction molecules. Alpha 1- and alpha 2-adrenergic receptors are single polypeptides with molecular mass 85,000 Da and pI 4.6. Beta 1- and beta 2 adrenergic receptors are single polypeptides with molecular mass 68,000 Da and pI 5.0. Muscarinic cholinergic receptors from a variety of tissues and species are single polypeptides with molecular mass 80,000 Da and pI 4.2. Proteolytic digestion and analysis of affinity-labelled adrenergic and cholinergic receptors indicates a striking similarity in the number and sizes of peptides produced. Topographical analysis of the receptors has shown that they have a similar membrane orientation with more than half of the protein exposed to the extracellular environment. Peptide-mapping studies of soluble and membrane-bound receptors suggest that the ligand-binding domain of adrenergic and cholinergic receptors is localized near the end of the protein that is exposed to the extracellular environment. The marked similarity between alpha- and beta adrenergic and muscarinic cholinergic receptor structure is perhaps not unexpected in light of the fact that these receptors interact with the same transmitters (in the case of alpha- and beta-adrenergic receptors) and/or with the same effector proteins in the membrane (stimulatory and inhibitory guanine nucleotide regulatory proteins, ion channels). Yet, depending on the tissue distribution of receptors and their effectors, this limited number of proteins can modulate dramatically different physiological effects. It may be that the differences in pharmacological specificity of ligand binding and the differences in receptor-effector interactions observed among adrenergic receptor subtypes and muscarinic cholinergic receptors are due to minor structural differences within the active sites of these proteins. Obviously, the real answers as to the extent of structural homology between the receptor classes will be derived from the amino acid sequencing of the purified proteins or from the cloning of the receptor genes and recently the genes coding for the beta-adrenergic receptor have been cloned and sequenced from human brain, hamster lung, and turkey erythrocytes. Comparison of the derived protein sequences reveals a high degree of structural homology between the avian and mammalian receptors with approximately 50% primary sequence identity and highly conserved secondary structure.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3034284 TI - The hormonal regulation of adenylate cyclase. AB - The regulation of adenylate cyclase by hormones and by GTP regulatory proteins was investigated in native membrane systems and in systems reconstituted from purified components. These studies can be summarized as follows. The stimulatory beta 1-adrenoceptor catalyses the activation of a complex between the GTP stimulatory protein GS and the catalytic unit C. The agonist-receptor complex can activate a few cyclase units in native membrane systems as well as in reconstituted systems. GS from turkey erythrocytes is functionally different from rabbit liver GS, the latter being more amenable to activation by guanyl nucleotides in the absence of hormone. The coupling between the beta 1 adrenoceptor GS and C is efficient when compared with the coupling obtained in native membrane systems. GTP/GDP exchange at the alpha S subunit requires the presence of the beta gamma subunits. A mechanism for the inhibition of adenylate cyclase by the inhibitory GTP regulatory protein Gi is suggested. PMID- 3034286 TI - The vertebrate glycine receptor protein. AB - Glycine is a major inhibitory neurotransmitter in the central nervous system. The postsynaptic receptor for this amino acid is a transmembrane ion channel protein which, after affinity purification on an antagonist column, contains three polypeptides of Mr 48,000, 58,000 and 93,000. Biochemical and immunological data show that these polypeptides have different functional properties and topologies with respect to the postsynaptic membrane. A model summarizing the currently available structural data is presented. PMID- 3034285 TI - The GABAA receptor and its antibodies. AB - The GABAA/benzodiazepine receptor has been purified to homogeneity from bovine and rat cerebral cortex. Under optimum conditions, the purified receptor has been shown to possess four distinct drug-binding sites, for GABA, benzodiazepine, barbiturate and Cl- channel gating classes of ligands. The receptor is a multi subunit membrane glycoprotein with an oligomeric size of 230,000 Da. It contains at least two subunits, alpha and beta, the first of which can be photoaffinity labelled with the benzodiazepine, flunitrazepam. Polyclonal and monoclonal antibodies have been raised to the native receptor and both have been used in an immunological characterization of the receptor protein in detergent extracts and likewise in purified preparations from bovine cerebral cortex. PMID- 3034287 TI - Expression and function of the human calcitonin/alpha-CGRP gene in health and disease. AB - Studies on the structure and expression of the rat and human calcitonin gene provide a remarkable example by which a combination of molecular, immunocytochemical and pharmacological techniques have led to the identification of a novel gene product, the calcitonin gene-related peptide, a peptide of unexpected tissue distribution and biological activity. Future studies at the molecular level will provide insight into post-transcriptional mechanisms by which a single gene can give rise to discrete mRNAs in a tissue-specific manner. The isolation and characterization of the CGRP receptor, and hence the site(s) and mechanism(s) of action of the CGRP family within the vasculature, will also add significantly to our understanding of molecular mechanisms involved in the modulation of cardiovascular function in man. Furthermore, the role and mechanism of action of the CGRP peptide family in the central and peripheral nervous systems remains to be elucidated. It is also apparent that measurement of circulating plasma levels of CGRP may be of diagnostic and prognostic value, providing information concerning the onset and progression of lung and thyroid carcinoma. Our analysis of the structure and expression of the calcitonin/alpha CGRP gene also demonstrates, for the first time, the molecular basis of large calcitonin secretion by lung carcinoma cells. Whether the expression of this gene in lung carcinoma cell-lines can truly be described as 'ectopic' is however questionable, in the light of recent immunocytochemical evidence which demonstrates that the lung is a major source of CGRP producing cells in the rat (see Springall et al., 1984). PMID- 3034288 TI - Modulation of neuronal MAO activity, 5-HT uptake and imipramine binding by endogenous substances in dog cerebrospinal fluid. AB - Addition of small amounts of dog cerebrospinal fluid (CSF) inhibited both type A and type B monoamine oxidase (MAO) in dog brain mitochondria. The inhibition was competitive with 5-HT as substrate, but non-competitive with beta phenylethylamine as substrate. Tricyclic antidepressants also exhibited competitive inhibition with type A MAO, but were non-competitive with type B MAO. The endogenous materials in CSF activate [3H]-imipramine specific, dose-dependent binding in dog brain preparations. The maximum number of binding sites (Bmax) increased, but the dissociation constant (Kd) was altered significantly in the presence of CSF. Addition of CSF induced a marked activation of uncompetitive [14C]-5-HT uptake in dog brain preparations. Moreover, there were reversibilities of the inhibition of MAO activity or of the activation of imipramine binding and 5-HT uptake by CSF substance after dilution experiment. These results indicate the possible presence of an endogenous psychotic drug-like substance in CSF. PMID- 3034289 TI - Effects of isoproterenol on active force and Ca2+ X calmodulin-sensitive phosphodiesterase activity in porcine coronary artery. AB - Relaxation of vascular smooth muscle following beta-adrenergic stimulation may result from reduction of the cytoplasmic Ca2+ concentration, reduction in the sensitivity of the contractile apparatus to Ca2+, or both. To help resolve these possibilities, we measured the extent of activation of Ca2+ X calmodulin sensitive phosphodiesterase in intact porcine coronary artery strips as a functional indicator of the cytoplasmic Ca2+ concentration. Both calmodulin stimulated phosphodiesterase activity and active force increased during K+ stimulation of coronary artery strips. Relaxation of K+-contracted artery strips following stimulus withdrawal was accompanied by rapid inactivation of Ca2+ X calmodulin-sensitive phosphodiesterase. The temporal relationship between isoproterenol-induced relaxation and inactivation of Ca2+ X calmodulin-sensitive phosphodiesterase was studied in both histamine- and K+-contracted tissues. Stimulation of strips with 10 microM histamine or with 44 mM K+ led to comparable increases both in active force and in calmodulin-stimulated phosphodiesterase activity. Thereafter, sustained contraction elicited by histamine was accompanied by a decrease in calmodulin-stimulated phosphodiesterase activity. Isoproterenol rapidly relaxed histamine-contracted strips and accelerated the decrease in phosphodiesterase activity. In contrast, sustained contraction in response to K+ was accompanied by a sustained elevation of calmodulin-stimulated phosphodiesterase activity. Isoproterenol treatment of K+-contracted tissues led to relaxation that was slow and incomplete, and it had very little effect on calmodulin-stimulated phosphodiesterase activity. We conclude that reduction of the cytoplasmic Ca2+ concentration is important for rapid relaxation of the coronary artery following beta-adrenergic stimulation. We cannot disallow the possibility that a decrease in the sensitivity of the contractile apparatus to Ca2+ is involved to some degree. PMID- 3034291 TI - Characterization of the beta 2-adrenoceptor-dependent adenylate cyclase of A431 epidermoid carcinoma cells. AB - In this study we characterize the beta 2 adrenergic dependent adenylate cyclase system of epidermoid carcinoma cells (A431). We show that the cells synthesize up to 130,000 [125I]-cyanopindolol binding sites per cell when freshly plated, a value which decreased to 40,000-50,000 receptors/cell within 24 hr. Production of this high number of receptors can be strongly inhibited by actinomycin D. We confirm and extend the fact that these beta-adrenoceptors are of the beta 2 subtype, using selective ligands, photoaffinity labeling with [125I]CYP-diazirine identified two protein subunits: p59 and p72, the beta 2-adrenoceptor dependent adenylate cyclase desensitizes with half-life of 2.2 +/- 0.3 min whereas the loss of [125I]CYP binding from the cell surface requires longer exposure times to the agonist, phorbol-12-myristate-13-acetate (PMA) has no effect on the desensitization process nor does it have any effect on the modulation of beta agonist affinity by guanyl nucleotides. Rather, PMA was found to stimulate adenylate cyclase activation by forskolin. We conclude that protein kinase C is probably not involved in the beta-adrenoceptor desensitization in this cell line. PMID- 3034290 TI - Differential inhibitory effects of auranofin on leukotriene B4 and leukotriene C4 formation by human polymorphonuclear leukocytes. AB - Auranofin (AF) is a newly introduced oral gold compound having antirheumatic properties, and its efficacy in the treatment of bronchial asthma is now under investigation. In this study, we examined the effects of AF on leukotriene (LT) formation by human polymorphonuclear leukocytes (PMNs) stimulated with the calcium ionophore A23187. AF inhibited LTC4 formation in a dose-dependent manner with an IC50 (concentration required to produce 50% inhibition of control) of 3.2 microM. In contrast, LTB4 formation was not prevented by AF at concentrations up to 6 microM, but it was reduced to 59 +/- 4% (mean +/- SE, N = 3) of control by an 8 microM concentration. As a next step, we explored the mechanisms of the differential inhibitory effects of AF using cell-free systems. When arachidonic acid (AA) and reduced glutathione (GSH) were used as substrates, AF inhibited LTC4 synthesis more effectively (IC50 = 14 microM) than LTB4 synthesis (IC50 = 100 microM). However, LTB4 and LTC4 syntheses from LTA4 were affected only slightly by AF within the concentrations tested (3-100 microM). These results in the cell-free systems indicate that the inhibition of LT formation was caused by a reduction of LTA4 synthesis and that the differential inhibitory effects can be ascribed to the higher Km value of glutathione S-transferase for LTA4 than that of LTA4 hydrolase in PMNs. In accordance with this hypothesis, LTC4 synthesis was more dependent than LTB4 synthesis on LTA4 concentrations within 25-100 microM, and AA-861, a 5-lipoxygenase inhibitor, caused similar differential inhibitory effects on the formation of LTs by intact PMNs. The inhibitory effect of AF on LT formation at physiological concentrations may play some role in the efficacy of this drug. PMID- 3034292 TI - Synergistic antiviral effect of xanthates and ionic detergents. AB - Xanthate compounds have been shown to exhibit antiviral activity against various DNA and RNA viruses under acidic pH conditions. It is now possible to utilize the unique broad range antiviral spectrum of these compounds under physiological pH conditions (pH 7.4) by simultaneous administration of certain ionic detergents. When used in conjunction with tricyclodecan-9-yl-xanthate (D609), sodium deoxycholate, sodium dodecylsulfate and certain fatty acids, which have no antiviral activity of their own, inhibit the replication of various DNA and RNA viruses (such as herpes simplex, vesicular stomatitis and Coxsackie B 4) in vitro at pH 7.4. Among saturated fatty acids of various chain lengths there was a marked size restriction in that the efficiency of undecanoic acid (11 C atoms) was three orders of magnitude greater than that of shorter (6 C atoms) or longer (18 C atoms) monocarbonic acids. Dose-response kinetics revealed a synergistic interaction between the xanthate and the monocarbonic acid. A dose that inhibited the replication of herpesvirus by a factor of 1000 still permitted mitotic activity in uninfected growing control cultures. PMID- 3034293 TI - [The role of structural elements of ColE1-related plasmids in replication control]. AB - Effect of deletions downstream from the replication origin on copy number of ColE1 related (pBR322 derived) plasmids has been studied. Along with main control elements (RNAI of defined secondary structure, the repressor protein, effectiveness of promoter upstream of gene of RNA primer) replication is influenced by transcription level propagated into the replicon region from promoters even in distal parts of plasmid. Structure of the region adjacent to the ori is important for startpoint recognition specificity, so that deletion of this region reduces the effectiveness of replication. PMID- 3034295 TI - Inclusion body myositis presenting as treatment-resistant polymyositis. AB - Inclusion body myositis (IBM) has been viewed as a distinct and rare form of inflammatory myopathy. Previously reported findings from series of IBM patients have suggested that clinical and pathologic features are present which readily distinguish it from idiopathic polymyositis. We report 4 cases of IBM presenting clinically and pathologically as polymyositis, each of which was refractory to therapy. Our data suggest that IBM may be a more common and heterogeneous form of inflammatory myopathy than has been previously suggested. Furthermore, IBM may be clinically and electrophysiologically indistinguishable from polymyositis. Reasons for failing to recognize IBM by pathologic studies appear to include: the skip lesion nature of the pathologic findings, failure to examine tissues by electron microscopy, and a low level of suspicion or lack of recognition. Because of its insidious clinical course and its failure to respond to immunosuppressive therapy, IBM may be an important variant of treatment-resistant polymyositis. PMID- 3034294 TI - [DNA-duplexes with phosphoamide bond: interaction with restriction endonucleases EcoRII and SsoII]. AB - We studied the interaction of EcoRII and SsoII restriction endonucleases with synthetic DNA duplexes, containing 3'N----5'P and 3'P----5'N phosphoamide internucleotide bonds in one of the cleavage points. Enzymatic hydrolysis of the modified strand of the duplexes is blocked in all cases. The presence of phosphoamide bonds was found to reduce the rate of cleavage of the natural strand by EcoRII and to have no influence in case of SsoII. Properties of the EcoRII endonuclease complex with its substrate, containing non-cleavable 3'N----5'P internucleotide bonds in each cleavage point, were examined. In the presence of Mg2+ ions the equilibrium association constant of the enzyme-substrate complex is 3-fold reduced, and the dissociation rate constant of the complex is increased by 1.5 times. PMID- 3034296 TI - Human recombinant interleukin-1 beta stimulates glycosaminoglycan production in human synovial fibroblast cultures. AB - Human recombinant interleukin-1 beta (rIL-1 beta) stimulated glycosaminoglycan (GAG) production in human synovial fibroblast cultures. A dose-dependent increase in GAG production was found, to a maximum of 500%. Increase was detected at doses as low as 1 pg/ml of rIL-1 beta, reached a maximum at 10-100 pg/ml, and was apparent 10 hours after addition of rIL-1 beta. Stimulation of GAG was always accompanied by increased accumulation of prostaglandin E (PGE) in culture media and by increased collagenase production in approximately one-half the experiments. Indomethacin (5 micrograms/ml) completely inhibited PGE stimulation by rIL-1 beta, but only partially inhibited that of GAG overproduction and had no effect on collagenase production. Hydrocortisone (2 micrograms/ml) inhibited stimulation of all 3 parameters. Stimulation of hyaluronate in synovial cultures prevailed over that of sulfated GAG, which occurred to a lesser extent. Our results support earlier suggestions that interleukin-1 is a major active mononuclear cell factor that is capable of inducing profound changes in connective tissue cell function. PMID- 3034297 TI - Receptor-mediated binding of leukocyte elastase by chondrocytes. AB - Studies on the effect of leukocyte elastase on the metabolism of chondrocytes in culture have demonstrated that these cells possess a specific cell surface receptor for leukocyte-derived elastase. Purified elastase from rabbit and human leukocytes is capable of modulating the metabolism of the cell by causing a marked decrease in both proteoglycan and protein biosynthesis. Addition of 125I labeled elastase to chondrocytes maintained in suspension culture has shown that binding occurs, and that it is saturable and is inhibited by the addition of unlabeled enzyme. We ascertained that the active site of the enzyme was necessary for binding to the chondrocyte, since phenylmethylsulfonyl fluoride-inactivated leukocyte elastase failed to bind. Pancreatic elastase had only a slight affinity for the receptor, whereas trypsin and bovine serum albumin failed to bind to any significant extent. Autoradiographic studies and the use of inhibitors of endocytosis, such as dansyl cadaverine, confirmed that endocytosis of elastase was the secondary event after cell binding. PMID- 3034299 TI - Reproductive toxicity studies of rentiapril. AB - (2R,4R)-2-(o-Hydroxyphenyl)-3-(3-mercaptopropionyl)-4- thiazolidinecarboxylic acid (rentiapril, SA 446), an orally active inhibitor of angiotensin converting enzyme, was examined for effects upon general reproductive performance, for embryofoetal toxicity and for peri- and postnatal toxicity in the rat at dosages of 0, 20, 100 and 500 mg/kg/d. Embryofoetal toxicity was also examined in the New Zealand White rabbit at dosages of 0, 1, 2 and 4 mg/kg/d. The compound was administered by gastric intubation. Prolonged treatment at 100 and 500 mg/kg/d during the fertility study was associated with some slight depression of body weight gain of males. Body weight gain of females during gestation was significantly depressed at 500 mg/kg/d. There was salivation in both sexes at 500 mg/kg/d and also in males receiving 100 mg/kg/d. Following this prolonged treatment in the fertility study. Fo male and female kidney weights were increased at all dosages. Although there was no obvious effect upon fertility there was an increased incidence of total litter loss at 500 mg/kg/d and mean pup weights to day 21 post partum were reduced at this dosage and at 100 mg/kg/d with delays in the attainment of some of the developmental landmarks. In the rat treatment at 500 mg/kg/d from day 7 to 17 of pregnancy did not adversely effect embryofoetal development. Subsequent development and reproductive performance of the F1 offspring was also unimpaired. During this treatment period signs of salivation were seen at 500 mg/kg/d. Slight retardation of maternal body weight gain was noted at 500 mg/kg/d and at 100 mg/kg/d but not at 20 mg/kg/d.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3034298 TI - General pharmacological properties of the potent angiotensin converting enzyme inhibitor rentiapril. AB - Pharmacological properties of (2R,4R)-2-(o-hydroxyphenyl)-3-(3-mercaptopropionyl) 4- thiazolidinecarboxylic acid (rentiapril, SA 446), a potent inhibitor of angiotensin converting enzyme, were examined. SA 446 given intravenously lowered blood pressure in anesthetized rats and rabbits dose-dependently, while it had no remarkable effect in conscious rats even at a dose of 100 mg/kg orally or intravenously. SA 446 had no effect on heart rate in anesthetized rabbits up to 100 mg/kg i.v. In anesthetized dogs, SA 446 at a dose of 1 mg/kg i.v. produced decrease in blood pressure accompanied with coronary and vertebral arterial vasodilations, but gave no change in heart rate. SA 446 had no influence on respiration of anesthetized rabbits at doses up to 100 mg/kg i.v. The drug showed no effect on contractile force or beating rate of isolated guinea pig atria at concentrations up to 10(-4) mol/l, but caused vasoconstriction of isolated rabbit ear vessels at a 0.1 ml dose of 3 X 10(-2) mol/l solution or more and relaxation of isolated rat thoracic aortas at concentrations over 3 X 10(-4) mol/l. SA 446 had no influence on beta 1- or beta 2-adrenoceptors of isolated guinea pig atria and tracheas, respectively. Neither was a calcium antagonistic action observed. SA 446 did not influence the central nervous system, neuromuscular transmission, spontaneous movement or contractile response to several agonists of isolated smooth muscles, secretion of digestive glands, blood coagulation, hemolysis or blood glucose level, and it showed no muscle relaxant, antispasmodic or local anesthetic effect.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3034300 TI - Ploidy of endothelium in high-grade astrocytomas. AB - To determine the ploidy and proliferative activity of the endothelium in high grade astrocytomas, the nuclear DNA content of 11 high-grade astrocytomas, including two gliosarcomas, was measured by cytophotometry. This technique allowed comparison of the endothelial population with the astrocytic population. In all cases, the endothelium was diploid, with an average of 24.6% of cells in the S + G2M phases of the cell cycle. In contrast, the astrocytic population displayed marked DNA abnormalities. The two gliosarcomas had a marked difference in proliferative activity of the endothelium with 4% and 40% of cells, respectively, in the S + G2M phases. These data indicate that the vast majority of endothelial cells compromising the vascular hypercellularity observed in high grade astrocytomas are in the normal cell cycle whereas in many cases the malignant astrocytes are not. The nuclear DNA content of gliosarcomas appears to be similar to other high-grade astrocytomas. PMID- 3034301 TI - Role of temporal order and odor intensity in taste-potentiated odor aversions. AB - The role of the temporal order of odor and taste was studied in two experiments, and a third experiment studied the role of odor intensity in flavor-toxicosis conditioning with thirsty rats licking water spouts in a "wind tunnel." In all experiments, odors and tastes were presented for 2 min to rats, and 30 min later, a toxin (lithium chloride) was intubated. In Experiment 1, an odor was presented 90 s before, during, or 90 s after a taste to independent groups. Experiment 2 was a within-subjects partial replication of the first. Each rat was presented with one odor, then a taste, then a second odor with each stimulus separated by 45 s. The results of Experiments 1 and 2 indicated that (a) odor alone is not associated with illness under our conditions, (b) presenting an odor and a taste at the same time potentiates the odor component so that it is associated with illness, (c) 45-s and 90-s intervals between odor and taste eliminate potentiation, and (d) taste and odor interact asymetrically; that is, odor has little affect on the development of taste-illness associations. In Experiment 3, an odor and a taste were presented simultaneously, and odor intensity varied. As odor intensity increased, the strength of the taste-potentiated odor aversion increased, whereas the aversion to the taste remained constant. However, even at the highest intensity, odor presented in the absence of taste did not result in odor aversions. PMID- 3034302 TI - Preexposure and extinction effects of lithium chloride induced taste-potentiated aversions for spatially contiguous auditory food cues in rats. AB - Taste potentiated illness-induced aversions for noisy food were studied in Sprague-Dawley rats. The rats ate from receptacles containing salty food and a contiguous tone produced by speakers under the food followed by lithium chloride injections. In preference tests, the rats then avoided noisy food in favor of quiet food followed by extinction and spontaneous recovery of the auditory aversion over repeated nonreinforced trials. Other rats were given either 4 or 10 days of exposure to the noisy food prior to taste-toxicosis treatment. None of these rats subsequently avoided noisy food. The importance of spatial contiguity and methodological variation in associating nongustatory food cues with illness is discussed. PMID- 3034303 TI - The central representation of an aversive event maintains the opioid and nonopioid forms of analgesia. AB - Exposure to an aversive event, such as shock, can elicit either an opioid or nonopioid analgesia in rats. We suggest that the central representation of an aversive event in working memory activates both forms of analgesia. We formalize this basic hypothesis by coupling it with a current model of animal learning and memory, SOP (Wagner, 1981). SOP is designed to capture the standard operating procedures that govern memory systems. Our application of SOP suggests that manipulations which disrupt the maintenance of information in working memory should alter the magnitude and time course of analgesia. Three experiments are reported that support our proposal. Experiment 1 showed that analgesia decays more rapidly if the representation of the aversive event is displaced from working memory by presenting a postshock distractor. Experiment 2 demonstrated that the postshock distractor alters the magnitude and time course of both the opioid and nonopioid forms of analgesia. Experiment 3 demonstrated that pharmacologically disrupting working memory, by administering a high dose of pentobarbital, prevents mild shock from inducing a strong change in pain reactivity. Implications of the results are discussed. PMID- 3034304 TI - Cessation of male rat copulatory behavior using illness as punishment: facilitation with a novel odor. AB - Two experiments were conducted to examine learned copulatory avoidance in male rats. One group of males was presented with receptive females that had been sprayed with a 2% almond solution, and the other group was presented with nonalmond odorous, receptive females. Following each test, males were made ill with lithium chloride (LiCl) by intragastric intubation or intraperitoneal injection. Results showed that male rats presented with almond-odorous females developed significant avoidance of copulatory behavior. Conditioning in males exposed to receptive females without the almond odor developed little, if any, avoidance. In Experiment 2, it was found that route of LiCl administration was not a factor in the results. PMID- 3034305 TI - Primary hepatitis A virus infection in developing countries: decline of maternal antibodies and time of infection. AB - In developing countries, HAV seems to be responsible for a widespread, inapparent and protective infection during early childhood. This report emphasizes early infection and its relationship to protection by passive immunity from maternal antibody in a highly endemic area such as Somalia. Our result show that HAV infection in Somalia primarily occurs during the first 4 years of life (4 months to 4 years). Cases are infrequent in the first 3 months due to passive immunity secondary to maternal antibody (cord-blood and colostrum anti-HAV). As the level of protection declines, the rate of acute infection rises as determined by the presence of IgM-specific anti-HAV. PMID- 3034306 TI - Comparison of indirect immunofluorescence and radioimmunoassay for detecting antibodies against hepatitis A virus of the IgG and IgM classes. AB - Indirect Immunofluorescence test (IIF) and Radioimmunoassay (RIA) were compared for determining anti-Hepatitis A virus (HAV) IgG and IgM. 142 sera were tested for anti-HAV IgG and 16 for anti-HAV IgM, both with IIF and RIA techniques. The correlation between the results reached 98.6% for anti-HAV IgG and 93.75% for anti-HAV IgM detection, confirming the specificity and sensibility of IIF. PMID- 3034307 TI - Escape rate of pertechnetate from lumbar CSF of man. PMID- 3034308 TI - Chronic ethanol and neuronal membranes effect on adenylate cyclase. PMID- 3034309 TI - Algorithms for morphometric measurements on cancer cells in electron microscopy. Pilot tests. AB - Automated procedures were designed for handling quantitative data derived from a morphometric study of neoplastic cells on electron micrographs. Pilot studies were carried out on 3 breast carcinomas. Each tumor cell was coded by the sequence of 30 stereological parameters that described the general organization of subcellular constituents. Wide standard deviations, as large as 90% of the mean, were observed in some parameters. The practical application of correlation analysis between stereological parameters turned out to be of limited value, since only parameters related to the same subcellular structure showed correlation. PMID- 3034310 TI - Correlation between morphometrical parameters and disease-free survival in ductal breast cancer treated only by surgery. AB - A combination of quantitatively evaluated morphological parameters and of conventional prognostic indicators has been used to study 19 cases of invasive ductal breast carcinoma from patients treated only by surgery, later developing recurrences or metastases. This set of patients not treated with adjuvant therapy (radiotherapy, cytostatic or hormonal therapy), was selected from 350 consecutive breast cancers which had been treated with surgical therapy. The aim was to investigate whether the morphometrical features are correlated with disease-free survival. Of the single features, the mitotic activity index (MAI) is most strongly correlated with prognosis. Ten of the 19 patients had an MAI value above 9, and all of them recurred within 18 months. In contrast, of the 9 other patients with low mitotic rate (MAI below or equal to 9), none recurred within 24 months. Further, a correlation exists between disease-free survival, mean and standard deviation of nuclear and nucleolar area, and tubular component of the tumors. There is no correlation between nuclear form factors and recurrence. The multivariate prognostic score is also significantly correlated with recurrence, but not as strongly as in other publications. This is obviously due to the blurring influence of the lymph node status, which was not significantly correlated with the prognosis. Thus, in this small set of patients not treated with any adjuvant therapy, the results of morphometric analysis are in agreement with earlier data and emphasize the prognostic significance of quantitative microscopical analysis in breast cancer. PMID- 3034311 TI - Evaluation of carcinoembryonic antigen and periodic acid-Schiff stains in the diagnosis of serous and mucinous ovarian tumors: morphometrical study. AB - 108 serous and 104 mucinous neoplasms of the ovary, classified according to the criteria of the World Health Organization, were studied with periodic acid-Schiff (PAS) method for mucin and glycogen, and with immunoperoxidase method for carcinoembryonic antigen (CEA). The amount of epithelium, the number of mitoses, and the staining positivities were quantitated morphometrically and the parameters were evaluated using the discriminant analysis. In general, CEA scores were higher in mucinous than in serous tumors and higher in malignant tumors than in their benign or borderline counterparts. High PAS positivities were found both in serous and mucinous borderline lesions. Mitotic activity correlated with the malignancy of the tumors. The best discriminator, however, was the amount of epithelium. The results show that simple morphometry with standard stains is an efficient method in the first-line classification of these tumors. The quantitation of CEA and PAS stains may be helpful in cases in which histological criteria do not allow definite classification. PMID- 3034312 TI - Clinical, histological and morphometrical parameters in small cell carcinoma of the lung: correlation with survival. AB - 41 cases of small cell carcinoma of the lung were studied in order to analyze the relationship between survival and 'performance status', extent of the disease, response to therapy, histological subtypes according to WHO (1981) and some morphometrical parameters (nuclear area and form factors). Performance status, extent of the disease and response to therapy are significant prognostic indicators, while histological subtypes and morphometrical parameters have no significant correlation with prognosis. PMID- 3034313 TI - [Reappraisal of the treatment of age-dependent epileptic encephalopathy with ACTH]. PMID- 3034314 TI - [An atypical case of group A xeroderma pigmentosum]. PMID- 3034315 TI - [Lipid composition of human malignant brain tumors]. AB - Malignant transformation is characterized by the uncontrolled proliferation of cells. And changes in the composition of glycolipids, cell surface component which may be involved in regulation of cell growth, were often observed in the malignant transformation. In this study, cholesterol, lipid-bound phosphorus, cerebroside, sulfatide and ganglioside were quantitated in the tissue of 20 human malignant brain tumors (malignant glioma, 8; low grade glioma, 4; metastatic tumor, 7; malignant meningioma, 1). As compared with normal brain, all tumor tissue contained lower cholesterol, sialic acid, cerebroside and sulfatide. Metastatic brain tumor or glioma showed characteristic patterns in the content of ganglioside, cerebroside and sulfatide respectively. The ganglioside patterns of metastatic tumor or glioma contained a greater proportion of structurally simpler gangliosides than normal brain. And in metastatic tumor, GM3 was a major ganglioside. On the contrary, glioma had increased proportion of GM3 and GD3 gangliosides. High grade glioma such as Grade 3-4 contained higher proportion of GM3 and GD3, whereas low grade glioma (Grade 1-2) contained less proportion of GM3 and GD3. PMID- 3034316 TI - The pharmacokinetics of enalapril in hospitalized patients with congestive heart failure. AB - The pharmacokinetics of the converting enzyme inhibitor, enalapril, were studied in an open, randomized, balanced crossover design in 12 hospitalized patients with stable, chronic congestive heart failure (CHF). Enalapril maleate is a prodrug requiring in vivo hepatic esterolysis to yield the active diacid inhibitor enalaprilat. CHF results in changes in regional blood flow that may affect the gastrointestinal absorption, hepatic hydrolysis and renal excretion of enalapril and enalaprilat. In order to evaluate the pharmacokinetics of enalapril in CHF, the following treatments were given: enalapril maleate 10 mg orally, enalapril maleate 5 mg intravenously and enalaprilat 5 mg intravenously. Each dose was followed by a 72 h period with frequent blood sampling and fractionated urine collection for the radioimmunoassay of enalaprilat, before and after sample hydrolysis. Mean absorption for the oral dose was 69%, hydrolysis 55%, bioavailability 38%, urinary recovery 77% and estimated first-pass effect 10%. The results were compared with available data in normal subjects. After oral administration of 10 mg enalapril maleate, the extent of absorption, the degree of hydrolysis and the bioavailability in CHF patients appear to be similar to those in normals with differences less than 10%. The rate of absorption and hydrolysis appear to be slightly slower in CHF. The serum concentrations of enalaprilat were consistently greater in CHF and maximal concentrations were reached at 6 h in CHF as compared to 4 h in normal subjects. We conclude that the presence of CHF does not appreciably alter the pharmacokinetic behaviour of enalapril. The observed differences may be associated with age as well as the disease state. PMID- 3034318 TI - Pharmacokinetics and biological effects of captopril and hydrochlorothiazide after acute and chronic administration either alone or in combination in hypertensive patients. AB - The pharmacokinetics of free unchanged captopril, total captopril and hydrochlorothiazide (HCTZ) were investigated in three groups of patients with moderate essential hypertension and normal renal function on the first and on the 45th days of an oral treatment with either captopril (50 mg once daily, n = 7) or HCTZ (25 mg once daily, n = 10) or their combination captopril 50 mg + HCTZ 25 mg once daily, n = 8. Simultaneously, the effects of the three treatments on plasma converting enzyme activity (PCEA) and plasma renin activity (PRA) were measured. Elimination half-lives of total captopril after acute (7.8 +/- 2.3 h) or chronic (7.0 +/- 0.5 h) captopril dosing were similar to those of HCTZ after acute (6.5 +/- 1.0 h) or chronic (8.0 +/- 2.5 h) HCTZ dosing. Elimination half-life of free unchanged captopril was 0.81 +/- 0.09 h after acute and 0.96 +/- 0.03 h after chronic captopril dosing. Addition of HCTZ to captopril induced no major change in free and total captopril pharmacokinetic parameters, in PCEA inhibition and in PRA increase (as they were determined after captopril alone) on acute as well as on chronic treatment. Addition of captopril to HCTZ induced no major change in HCTZ pharmacokinetic parameters and in PRA increase compared with those determined after HCTZ alone. Chronic treatment with captopril + HCTZ resulted, like chronic captopril treatment, in no accumulation of captopril, in no significant modification of free and total captopril pharmacokinetic parameters and of PCEA inhibition (as determined after acute administration) but led to a significant enhancement of the acutely induced increase in PRA. Chronic treatment with captopril + HCTZ resulted, like chronic HCTZ treatment, in no accumulation of HCTZ, in no significant modification of HCTZ pharmacokinetic parameters (as determined after acute administration) but also led to a significant enhancement of the acutely induced increase in PRA. Thus, the longer duration of the antihypertensive action of captopril + HCTZ as compared to that of captopril cannot be ascribed to a pharmacokinetic or biological interaction between captopril and HCTZ. PMID- 3034317 TI - Evolution of diuretics and ACE inhibitors, their renal and antihypertensive actions--parallels and contrasts. AB - The emergence of diuretic drugs and angiotensin converting enzyme (ACE) inhibitors ranks amongst the major therapeutic advances of modern medicine. The discovery of these drug groups arose largely by chance, yet each has dramatically influenced the treatment of congestive cardiac failure and arterial hypertension. The central role which diuretics have had in the management of both oedema and hypertension hinges on their ability to induce a net renal excretion of solute and water by selective interference with either active or passive ion transport processes in different segments of the nephron. Irrespective of sites of action, the continued antihypertensive action of diuretics is characterized by a reduction in plasma volume and extracellular fluid (ECF) volume that lasts for as long as the diuretic is given. The mechanism of this effect remains unclear but may involve autoregulatory reactions that leave cardiac output unaltered but maintain a sustained reduction in total peripheral resistance. ACE inhibitors also lower blood pressure by decreasing total peripheral resistance, leaving cardiac output, plasma volume and ECF volume unchanged. The detailed way these haemodynamic changes are achieved remains unknown but inhibition of converting enzyme present not only in the kidney but also in many extrarenal tissue sites, appears important. In both hypertension and cardiac failure, however, the kidney acts as a key target organ for ACE inhibitors. The increased renal vascular resistance and inappropriate renal salt excretion are reversed with enhanced renal blood flow and saluresis. Both angiotensin II (AII) and vasopressin mediated contraction of glomerular mesangial cells is inhibited, making glomerular filtration more efficient. Reduced aldosterone secondary to blockade of AII formation contributes to saluresis whilst encouraging positive potassium balance. ACE inhibition also impairs breakdown of kinins which may contribute to intrarenal and peripheral vasodilation either on their own or via release of prostaglandins and other vasoactive substances. The hypotensive actions of diuretics are potentiated by ACE inhibition primarily through blockade of AII formation and prevention of secondary aldosteronism. In combination, these drugs permit low doses to be used because of their synergistic effects. Caution has to be exercised whenever ACE inhibition is used, without and especially with diuretics, in the management of renovascular hypertension and other low-perfusion states. In these circumstances, AII plays an important autoregulatory role in preserving glomerular filtration through an increase in post-glomerular resistance.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3034319 TI - Use of a tritiated thymidine suicide technique in the study of the cytotoxic drug response of cells located at different depths within multicellular spheroids. AB - A technique using 'tritiated thymidine suicide' has been established as a means of studying the response to cytotoxic drugs of cells at different depths within multicellular tumour spheroids. Because of the characteristic spatial arrangement of cycling cells (mostly in the outer regions) and non-cycling cells (mostly at the inner regions) of spheroids, cells surviving after long term (24 h) exposure of spheroids to high doses of 3HTdR will be those located furthest from the surface. By comparing the drug response of cells from 3HTdR pre-treated and untreated spheroids, the individual response of total cells, cells near to the surface and cells lying deeper within the viable rim of spheroids can therefore be deduced. In this study, large spheroids of about 800 micron in diameter of a mouse mammary cell line, EMT6/Ca/VJAC, and of a human small cell lung cancer cell line, POC, have been used. Using clonogenic assay, the response of these two cell types to adriamycin (ADM), nitrogen mustard (HN2), CCNU and vincristine (VCR) (POC only) were measured. The preliminary part of this study has confirmed that the cells killed are those which incorporate 3HTdR during the DNA synthesis period; the cells killed are mainly located in the outer regions of spheroids i.e. surviving cells are mostly located in the inner part of the viable rim and 3HTdR pretreatment does not sensitise surviving cells to subsequent cytotoxic drug treatment. Results from large EMT6 spheroids agree with our previous findings (obtained using a selective disaggregation method) that cells in the outer regions of spheroids are more sensitive to ADM and HN2 than cells in the inner regions whilst the opposite is true for CCNU. For POC spheroids, cells in the outer region of spheroids are more sensitive to ADM and VCR than cells in the inner region whilst a reverse trend is seen for the response to CCNU. The response to HN2 is similar at all depths. Amongst the factors governing the response of cells in spheroids to cytotoxic drugs, the responses to ADM and VCR are thought to be largely dictated by cell cycle distribution and limited drug penetrability, whilst for HN2 the response may be determined by the factor of cell cycle distribution. For CCNU, we believe that the cellular response is largely dependent upon microenvironmental factors prevailing within spheroids. PMID- 3034322 TI - Response of polymorphonuclear leukocytes to topical leukotriene B4 in healthy and psoriatic skin. AB - The in situ infiltration of polymorphonuclear leukocytes following the topical application of leukotriene B4 to the clinically uninvolved skin of psoriatic patients and the skin of normal controls has been quantified using elastase as a marker enzyme. The magnitude of this response in the psoriatic patients proved to be significantly diminished compared with the response in the controls. The time course of the response was similar in both groups, maximum elastase activity being seen about 18 h after application of the LTB4. PMID- 3034321 TI - The effect of cycloleucine on SFV A7(74) infection in mice. AB - Cycloleucine (CL), a non-metabolizable amino acid analogue, was found to reduce thymus and spleen weights in Semliki Forest virus (SFV) strain A7(74) infected and control mice. The maximum effects were seen when three daily doses of CL were given to mice 24 h after an SFV A7(74) infection. In these mice thymus atrophy led to abolition of thymus dependent immune responses and changes in the pathological features of the viral infection--the most striking feature being prevention of demyelination. In addition virus titres in the brains of CL treated infected mice were increased and prolonged. These results show that demyelination following an SFV A7(74) infection is not a result of direct virus action, but of a T-cell mediated mechanism. PMID- 3034320 TI - The effects in the excess administration of prostaglandin E1 on the indicator enzymes of organella membrane in gastric mucosa. AB - Administration of excess vitamin A to rats causes gastric ulceration. In this study the effects on the gastric mucosa of excess vitamin A and excess prostaglandin E1, alone and in combination, were studied. Prostaglandin E1 protected against ulceration by vitamin A. Vitamin A labilized marker enzymes from four different membrane systems, namely those of the lysosomes, mitochondria, endoplasmic reticulum and plasma membrane, whereas only the effect on lysosomes was prevented by prostaglandin E1. Indeed, the prostaglandin alone labilized the enzymes from plasma membrane and endoplasmic reticulum and also damaged mitochondrial membranes. Both vitamin A and prostaglandin E1 caused a reduction in the total number and an increase in irregularly-shaped mitochondria in the parietal cells and produced dilation of the endoplasmic reticulum in both parietal and chief cells. It is noteworthy that prostaglandin E1 effectively prevents ulceration by vitamin A despite the extent to which it damages these membrane systems. These findings lend support to the hypothesis that vitamin A ulceration of the gastric mucosa is mediated via release of lysosomal enzymes, following damage to the lysosomal membranes. PMID- 3034323 TI - Plasma renin activity and cAMP in termination of pregnancy induced by sulprostone. AB - Plasma renin activity (PRA), plasma concentrations of cyclic adenosine-3',5' monophosphate (cAMP) and the serum levels of oestradiol, progesterone, chorionic gonadotrophin, potassium, sodium and calcium were estimated in connection with termination of pregnancy induced in 23 women by sulprostone, a derivative of prostaglandin E2. A rapid decrease in PRA was observed during and after infusion of 1000 micrograms or 1500 micrograms sulprostone. PRA returned to the initial values at 24 h after drug administration. The lowest levels of PRA were 36% and 51% of the initial values (P less than 0.001) in the two drug concentration groups. No significant changes were observed in the control group. The value for plasma cAMP did not correspond completely to the changes in PRA. Serum concentrations of oestradiol, progesterone and chorionic gonadotrophin showed a gradual decrease during drug administration. The changes in serum potassium, sodium and calcium were minimal. The decrease in PRA associated with sulprostone is very surprising. Our finding elucidates in part the still rather poorly known mechanisms and function of renin-angiotensin system in pregnancy. PMID- 3034324 TI - Serial urinary cyclic guanosine monophosphate measurements in the assessment of response to treatment in epithelial ovarian cancer. AB - Serial measurements of urinary cyclic guanosine monophosphate (cGMP) were performed in 47 patients with epithelial ovarian cancer. In 30 patients, the pre chemotherapy cGMP level was above the range for normal controls (marker positive); the remainder were within the normal range. Marker positive patients demonstrated a 96% correlation of disease regression with a fall in marker level and a 75% correlation of disease progression with a rise in marker level. The corresponding correlations in the marker negative group were 41% and 39%. Marker positive patients also demonstrated a rise in marker before clinical recognition of disease progression in six of 11 instances whereas only one of nine instances in marker negative patients showed such a rise. Patients with evidence of static disease had normal and stable marker levels. PMID- 3034325 TI - The kinetics of the aerobic oxidation of ferrocytochrome c by cytochrome c oxidase in solvents of increased viscosity are partially diffusion controlled. AB - The steady-state spectrophotometrically determined initial velocity kinetics of the aerobic oxidation of ferrocytochrome c by cytochrome c oxidase were examined for effects of diffusion control in solvents of increased viscosity. Both glycerol/water and sucrose/water proved unsatisfactory as viscosogens due to weak competitive inhibition (Ki values of 2.6 M and 1.6 M, respectively). However, polyethylene glycol (PEG) was satisfactory as a viscosogen. The measured diffusion coefficient of ferrocytochrome c in PEG/water was shown to follow closely the Stokes-Einstein equation. In PEG/water mixtures at high ionic strength the minimum association rate constant (kmin = Vmax/(Km[EO]) is partly diffusion controlled with contributions from diffusion control and chemical activation control being about equal at 5 mPa X s, a viscosity that may be physiologically relevant. This finding can be interpreted to mean that cytochrome c oxidase is an enzyme that has evolved to approach its maximum efficiency. The steady-state kinetics were also examined at low ionic strength where multiphasic kinetics are exhibited. The effect of increased viscosity was exhibited over the whole experimentally accessible region indicating that there are effects due to diffusion control on both the high-affinity and low-affinity binding of ferrocytochrome c. Several models for diffusion control were examined and a comparison is made with other diffusion-controlled reactions of proteins. PMID- 3034327 TI - Modulation of Na+-H+ exchange by ethinyl estradiol in rat colonic brush-border membrane vesicles. AB - Prior studies by our laboratory have suggested that a relationship may exist between rat colonic brush-border membrane vesicular fluidity and Na+-H+ exchange. To further explore this possible relationship, in the present studies the effects of ethinyl estradiol (17 alpha-ethinyl-1,3,5-estratriene-3,17-beta-diol) administration subcutaneously (5 mg/kg body wt. per day) for 5 days, on rat colonic brush-border membrane fluidity and Na+-H+ exchange were examined. This treatment regimen has previously been shown to decrease the lipid fluidity of rat hepatic and rabbit small intestinal plasma membranes. In agreement with these prior studies, the present results demonstrate that this agent decreases the lipid fluidity of treated-rat colonic brush-border membranes compared to control membranes, as assessed by steady-state fluorescence polarization techniques using three different fluorophores. An increase in the cholesterol content and cholesterol/phospholipid molar ratio of treated-membranes appear to, at least partially, be responsible for the fluidity differences. Furthermore, examination of the kinetic parameters for amiloride-sensitive sodium-stimulated proton efflux in treated and control membrane vesicles, utilizing the pH-sensitive fluorescent dye, Acridine orange, revealed that ethinyl estradiol administration decreased the Vmax for this exchange mechanism, expressed in arbitrary fluorescence units, by approx. 25% but did not influence its Km for sodium. These data, therefore, lend further support to the contention that alterations in fluidity may modulate Na+-H+ exchange in rat colonic brush-border membrane vesicles. PMID- 3034326 TI - Pre-steady-state reduction kinetics of QH2:cytochrome c oxidoreductase and the Q pool: evidence for a special quinone not in rapid equilibrium with the Q-pool. AB - The pre-steady-state kinetics of the reduction of the prosthetic groups of QH2:cytochrome c oxidoreductase in bovine heart submitochondrial particles were studied in relation to the kinetics of the Q-10 reduction, using duroquinol as substrate. The prosthetic groups, including semiquinone, were measured with EPR and low-temperature-diffuse reflectance spectroscopy, the samples being prepared with the rapid-freeze quench technique. For the determination of the redox state of ubiquinone in the pre-steady state the rapid chemical quench technique was used as an extension of the rapid-freeze quench technique, and Q-10 and QH2-10 were measured with reversed-phase HPLC after extraction with petroleum ether. Ubiquinone was reduced biphasically, 8% of total Q-10 (equal to 1 mol Q-10/mol cytochrome c1), being reduced within 5 ms, and the rest, the Q-pool, at a much lower rate. The initial rapid reduction of this special Q-10 was accompanied by rapid formation of Qi and rapid reduction of a large part of the cytochrome b 562. Both semiquinone formation and reduction of b-562 showed transient kinetics due to a contribution of the reaction pathway via centre o when the iron-sulphur cluster and cytochrome c1 were oxidised. The majority of the special quinol was located at centre i, probably bound, but also at centre o some bound quinol was formed. This was visible when antimycin was present, the antimycin-insensitive bound quinol being totally sensitive to myxothiazol. Myxothiazol alone accelerated the reduction of the Q-pool via centre i, but also the equilibration of cytochrome b-562 with the Q-pool. Antimycin drastically lowered the rate of reduction of the Q-pool and additionally seemed to block the rapid electron transfer from part of the Rieske iron-sulphur cluster to cytochrome c1. It is concluded that, during the pre-steady-state, cytochrome b-562 is not in equilibrium with the Q-pool and that the rate of equilibration is probably determined by the rate of dissociation of the special bound quinol from centre i. PMID- 3034329 TI - Gastric antisecretory activity of cycloheximide due to inhibition of protein synthesis. AB - Treatment of rats with cycloheximide 1 h before carbachol dose-dependently reduced the secretagogue-stimulated gastric acid secretion in pylorus ligated rats, and partially blocked carbachol- or histamine-induced activation of rat gastric (H+ + K+)-ATPase which includes translocation of reserve intracellular (H+ + K+)-ATPase into the apical membrane of the parietal cells and induction of a KCl pathway. Time-course studies showed that the drug was effective only when administered at least 30 min before the secretagogues. Puromycin showed the same effect as cycloheximide. Pulse labelling studies with [35S]methionine led to identification of two most actively synthesized polypeptides in rat gastric mucosa; the proteins of 38,000 and 14,000 molecular weight. The larger polypeptide was identified as rat pepsinogen. The identity of the smaller protein is not known yet. We suggest that synthesis of nascent polypeptide(s) is required for certain steps of the acid secretory process leading to the activation of the acid pump. PMID- 3034330 TI - Further evidence for coupling of sodium and proton movements in dog red blood cells. AB - Using 4,4'-diisothiocyanostilbene-2,2'-disulfonate (DIDS) and tributyltin the sodium transport pathway activated by shrinkage in dog red blood cells is shown to behave as expected for an electroneutral Na+/H+ exchanger. When the driving forces for sodium and protons are equal, flow through the pathway stops. Amiloride inhibits the shrinkage-induced Na+/H+ exchange. PMID- 3034328 TI - Separation of plasma membrane domains of calf thymocytes by affinity chromatography on ouabain-Sepharose. AB - Highly purified plasma membranes of calf thymocytes were fractionated by means of affinity chromatography on ouabain-Sepharose. By the method used two subfractions were obtained, one eluting freely from the affinity gel (MF1oua) and a second specifically retained by matrix-bound ouabain (MF2oua), with a total recovery of 95 per cent. Fractionation required the binding of matrix-bound ouabain to its plasma membrane receptor, i.e. (Na+ + K+)-ATPase. Increasing the temperature and binding time did not significantly alter the fractionation of plasma membranes into the two subfractions. Both plasma membrane subfractions separated by ouabain Sepharose were of plasma membrane origin, as revealed by the identical specific activities of several membrane bound enzymes, gamma-glutamyl transpeptidase, alkaline phosphatase and Mg2+-ATPase in unseparated plasma membranes and in both subfractions, and by the identical amounts of the cytoskeletal protein actin in unseparated plasma membranes and subfractions. The plasma membrane subfractions MF1oua and MF2oua showed different structural and functional properties. In SDS polyacrylamide gel electrophoresis polypeptides of 170, 150, 110, 94, 39, and 30 kDa were several-fold enriched in the adherent fraction, MF2oua. The phospholipid fatty acid composition of the plasma membrane subfractions proved to be different, as well. MF2oua contained significantly higher amounts of saturated fatty acids as compared to MF1oua. The specific activities of (Na+ + K+)-ATPase, Ca2+-ATPase and lysolecithin acyltransferase were highly enriched in the adherent fraction MF2oua, as compared to MF1oua. The data suggest that by the means of affinity chromatography on ouabain-Sepharose plasma membrane domains of the lymphocyte plasma membrane can be isolated, most probably implicated in the initiation of lymphocyte activation. PMID- 3034331 TI - Recent studies of DNA topoisomerases. PMID- 3034332 TI - The structure of a human neurofilament gene (NF-L): a unique exon-intron organization in the intermediate filament gene family. AB - We have cloned and determined the nucleotide sequence of the human gene for the neurofilament subunit NF-L. The cloned DNA contains the entire transcriptional unit and generates two mRNAs of approx. 2.6 and 4.3 kb after transfection into mouse L-cells. The NF-L gene has an unexpected intron-exon organization in that it entirely lacks introns at positions found in other members of the intermediate filament gene family. It contains only three introns that do not define protein domains. We discuss possible evolutionary schemes that could explain these results. PMID- 3034333 TI - Monkey (CV-1) mitochondrial DNA contains a unique triplication of 108 bp in the origin region. AB - A fragment of monkey kidney cells (CV-1) mitochondrial DNA (mtDNA) containing the origin of replication has been cloned and sequenced. The nucleotide sequence, 640 bp, extends from the coding sequence of phenylalanyl tRNA to the flanking sequence upstream of the origin region. A unique triplication of 108 bp, including the evolutionary conserved sequence CSB-3, was found. Comparison between the origin regions of monkey, human and mouse mtDNA is presented. PMID- 3034334 TI - [Study of lysozyme by a spin-label method in the 2-mm range]. AB - Two millimeter range ESR-spectroscopy was used to measure the values of magnetic resonance parameters of egg lysozyme samples modified with nitroxyl label 4 (iodine-acetamide)-2,2,6,6-tetramethylpiperidine-1-oxyl by hist-15 residue. It has been shown that in a lyophilic sample the spin label forms a hydrogen bond with the protein group and when the sample is moistened--with water molecules. Temperature changes of the pattern of ESR line are analysed. It is concluded that in the moistened samples within 230-320 K the nitroxyl fragment of the label gets engaged in anisotropic movement with preferable rotation around Z-axis of N-O. fragment with anisotropy coefficient 5 and mean correlation time tau congruent to 5 X 10(-7) divided by 5 X 10(-8) c. PMID- 3034335 TI - [Oscillations and trigger phenomena in fructose-2,6-bis-phosphate metabolism. A mathematical model]. AB - A mathematical model describing metabolism of fructose-2,6-bisphosphate (F2, 6P2), which is a powerful mediator in glycolysis, is investigated. The model takes into account inhibitory effect of F2, 6P2 and fructose-6-phosphate (F6P) on protein kinase, which phosphorylates the bifunctional enzyme fructose-6-phosphate 2-kinase/fructose-2,6-bisphosphatase. Such a mechanism of enzyme chemical modification in the presence of F2, 6P outflow from the F6P in equilibrium with F2, 6P2 cycle, caused by nonspecific phosphatases, can display trigger phenomena and sustained oscillations in F2, 6P2 metabolism and in the whole glycolytic system. The results obtained suggest that earlier models of the generation of glycolytic oscillations should be revised. PMID- 3034337 TI - [Reduction of cytochrome c adsorbed on liposomes]. PMID- 3034336 TI - [Effect of negative hydro-aero ions on the structure and functional properties of mitochondria]. AB - Studies were carried out of the physical mechanism of biological effect of natural stimulant of the living activity--light negative hydroaeroions. The latter are a hydroxyl separated from water ions and designated here as electrically noncompensated hydroxyl OH-. Unique effect of OH-, i.e. fast restoration and prevention of fine damage of native mitochondria (NM) at storage was revealed on mitochondria preparations by specific procedure, providing preservation of the ability of self-organization--formation of aggregated and denser packing--native mitochondria. This effect of OH- is conditioned by increased formation of native mitochondria aggregates and is concerned with better ADP phosphorylation and Ca2- release. The effect of the air current OH- is produced by water action presaturated with OH-. OH- induces in water cooperative structure formation preserved for many days, which is indicated by an increase of its heat capacity and decrease of surface tension. The principal factor in OH- effect both on mitochondria and water seems to be a decrease of local positive changes. It is of more advantage in NM water suspensions to order (structurate) NM with disordering water, which explains formation of NM stable aggregates. OH- effect supports the main function of the organism struggling with proton excess which destroys biological self-organization. Strengthened structural self organization under the effect of OH- found in NM seems to be of universal importance for biological macromolecules and membranes and is a primary effect of OH- on the living objects. PMID- 3034338 TI - [Effect of indoleacetic acid on the proton conductivity of biological membranes]. AB - The effect of indolylacetic acid (IAA) on proton conductivity of tylacoid membranes of isolated pea chloroplasts at pH 5.5-8.0 and of artificial phospholipid membranes at pH 7.5 were studied. IAA was shown to decrease the stationary proton gradient value and increase those of the dissociation constant and electron transport rate in chloroplasts, while in the artificial phospholipid membranes it increased the proton conductivity. The membrane lipid phase is supposed to be a possible result of phytohormone action, IAA transporting the protons according to the monomeric mechanism. PMID- 3034339 TI - [Calcium-blocking effect of nitro compounds in human platelets: correlation with changes in the cyclic guanosine monophosphate level]. AB - The effects of nitrates on Ca2+ increase and cyclic nucleotide content in human platelets were studied. Nitroglycerin, isosorbide dinitrate and sodium nitroprusside were found to inhibit the intracellular Ca2+ increase induced by the platelet activating factor, ADP and a stable thromboxane A2 analog--U46619. The inhibiting effect of sodium nitroprusside manifested itself at lower concentrations than those of nitroglycerin and isosorbide dinitrate. Nitroglycerin suppressed the Mn2+ entry into the cells and caused a 2-fold increase of the cGMP content which correlates with the calcium blocking activity. Methylene blue, a guanylate cyclase and glutathione reductase inhibitor, decreased the calcium blocking effect of nitroglycerin and its influence on the cyclic nucleotide content but failed to suppress the inhibitory effect of sodium nitroprusside. The data obtained suggest that the effects of nitrates on platelets are mediated by their influence on guanylate cyclase which leads to a cyclic nucleotide content increase and to a calcium blocking effect. PMID- 3034340 TI - [Biosynthesis and secretion of collagen in normal and SV-40 virus-transformed human embryonal fibroblasts]. AB - The biosynthesis and secretion of collagen proteins was studied in cultures of normal human embryo fibroblasts at different passages and growth stages as well as in cultures of human embryo fibroblasts transformed by oncogenic virus SV-40. It was found that normal fibroblasts maintain at a constant level the collagen synthesis throughout 20 passages, which is typical of proliferating and resting cells. Virus-transformed cells produce 3-4 times less collagen proteins on a per cell count. Normal and transformed fibroblasts do not differ in terms of total protein synthesis. Secretion of collagen and non-collagen proteins in transformed cell cultures appeared to be much lower than in normal cell cultures. Study of synthesized proteins by polyacrylamide gel electrophoresis showed that both types of cells secrete collagen proteins predominantly as polymers containing interchain S-S bonds of 3-helix molecules. Study of the protein-synthesizing activity of two polysomal fractions, i.e. membrane bound and free polysomes, isolated from the cells of both types in a cell-free system showed that membrane bound polysomes from transformed fibroblasts synthesize collagen much less actively in comparison with normal cells. However, in transformed cells free polysomes, in contrast with normal cells, are active participants of a cell-free collagen protein synthesis. PMID- 3034341 TI - [Purification and various properties of exonuclease from bacteriophage T4]. AB - Exonuclease A was isolated from bacteriophage T4-infected cells of E. coli. The molecular mass of the enzyme is approximately 42,000 Da, pH optimum is 7-8.5, pI is 4.05. The enzyme activity depends on Mg2+, the optimal concentration of Mg2+ being 1-5 mM. The enzyme splits one- and two-helical DNA in the direction of 3'-- -5' and is a deoxyribonuclease splitting 5'-deoxynucleotides. The enzyme shows a practically equal affinity for one and two-helical DNA. The Km value for one- and two-helical DNA is 10 +/- 1 and 11 +/- 1 pmole of chain DNA, respectively. The Vmax value for one- and two-helical DNA is 61 +/- 5 and 45 +/- 5 pmole of nucleotides per min. Exonuclease A may be used for preparing substrates for DNA polymerase T4 and Klenow fragment, i.e., during labeling of DNA at 3'-ends. PMID- 3034342 TI - Dehydroepiandrosterone sulfate stimulates collagenase synthesis without affecting the rates of collagen and noncollagen protein syntheses by rabbit uterine cervical fibroblasts. AB - The effects of dehydroepiandrosterone 3-sulfate (DHAS) and 17 beta-estradiol (E2) on collagen and noncollagen protein syntheses by rabbit uterine cervical cells were studied, and their effects on latent collagenase synthesis were compared. DHAS (1 X 10(-6) M) stimulated the synthesis of latent collagenase and did not affect the cell number and [3H]thymidine incorporation into DNA, whereas E2 had no effect on collagenase synthesis. On the other hand, neither DHAS (1 X 10(-6) M) nor E2 (1 X 10(-10)-1 X 10(-6) M) showed effects on collagen and noncollagen protein syntheses. These results suggest that the stimulative effect of DHAS on cervical ripening is mediated mainly by the stimulation of collagen catabolism, and that E2 does not concern the changes in the concentration of collagen and noncollagen protein in uterine cervix of the rabbit during pregnancy at term. PMID- 3034343 TI - Differential effects of prostaglandin F2 alpha and of prostaglandins E1 and E2 on cyclic 3',5'-monophosphate production and intracellular calcium mobilization in avian uterine smooth muscle cells. AB - The effects of prostaglandins (PGs) E1 (PGE1), E2 (PGE2) and F2 alpha (PGF2 alpha) on cyclic 3',5'-adenosine monophosphate (cAMP) production and intracellular Ca mobilization were examined in smooth muscle cells of chicken uterus grown in primary culture. At subnanomolar concentrations, both PGE1 and PGE2 significantly suppressed cAMP levels. However, at higher concentrations (0.1 100 microM), both agonists caused a dose-related increase in cAMP production. PGF2 alpha, on the other hand, had no effect on cAMP production. Forskolin (1-100 microM), which also stimulated cAMP production in a dose-dependent fashion, potentiated the effects of both PGE1 and PGE2. In digitonin-permeabilized uterine cells preloaded with 45Ca2+, the addition of PGF2 alpha caused a biphasic 45Ca2+ efflux. There was a small but significant 45Ca2+ release (10.0 +/- 1.5%) within 30 s (rapid phase), followed by a larger one (32.0 +/- 2.0%) within 5 min (slow phase). PGE2, at doses above 1 nM (which significantly increased cAMP accumulation), promoted 45Ca2+ sequestration. This action of PGE2 was observed as early as 1 min and was complete by 5 min. In addition, 0.001 nM PGE2 (a dose that was ineffective on 45Ca2+ mobilization) enhanced PGF2 alpha-induced 45Ca2+ mobilization from 22.5 +/- 5% to 57.0 +/- 3.5%. These results show that PGs of the E series have distinctly different effects on cAMP production and intracellular Ca mobilization. PGF2 alpha action may be linked directly to intracellular Ca mobilization, whereas the effects of PGE may be exerted at multiple sites depending on its local concentration. At low concentrations, its action may be mediated by the suppression of cAMP levels.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3034345 TI - Proton decoupled fluorine nuclear magnetic resonance spectroscopy in situ. AB - The efficacy of proton decoupling for enhancing the 19F nuclear magnetic resonance (NMR) signal-to-noise ratio and spectral resolution in the intact subject is demonstrated. A geometrically orthogonal cross-coil antenna configuration (Helmholtz pair, surface coil) is employed to provide 40 dB of isolation between the 19F observe and 1H decouple frequencies of 188 and 200 MHz, respectively. Further isolation is achieved through the use of high-quality notch filters on both observation and decoupling channels. Application of 19F-(1H) NMR spectroscopy to the study of 2-fluoro-2-deoxy-D-glucose metabolism in cerebral tissue in situ is presented. Significant improvements in sensitivity and resolution are obtained and result from both a collapse of the JFH multiple structure and a substantial positive nuclear Overhauser effect (NOE). To our knowledge, this is the first such demonstration of 1H decoupling in conjunction with 19F observation for study of the metabolism of a fluorinated compound in the living subject. PMID- 3034346 TI - The transmembrane helices of beef heart cytochrome oxidase. PMID- 3034344 TI - Heme-linked ionizations of myeloperoxidase detected by Raman difference spectroscopy. A comparison with plant and yeast peroxidases. AB - The pH-dependence of the oxidation state marker line v4 of human leucocyte myeloperoxidase is determined in the absence of chloride using Raman difference spectroscopy (RDS). A transition in the frequency of v4 with pK of 4.2 +/- 0.3 is found. The pK compares favorably with that previously determined by spectrophotometric titration and kinetic studies. The shift in v4 across the transition is -1.3 cm-1. The shift in v4 and other Raman marker lines indicates enhanced pi charge in the chlorin ring below the transition. The low frequencies of the oxidation state marker lines indicate that a structural change occurs near the chromophore, which results in the formation of a more pi-charge donating protein environment for the chlorin ring at low pH. The Raman results are discussed in terms of a proposed catalytic control mechanism based on charge stabilization of the energy of ring charge-depleted ferryl intermediates of the reaction with peroxide. The myeloperoxidase findings are compared with similar RDS results for ferrous horseradish peroxidase and ferric cytochrome c peroxidase. PMID- 3034347 TI - Cytochrome c chimerae from natural and synthetic fragments: significance of the biological properties. PMID- 3034348 TI - Detection of human papillomavirus types 6, 11, 16 and 18 in genital warts and cervical neoplasia. PMID- 3034349 TI - Ionic strength-dependent alterations of membrane structure of red blood cells. AB - Electron paramagnetic resonance (EPR) measurements using various fatty acid spin labels were performed on membranes of intact human erythrocytes at physiological, and at low ionic strength. In the case of spin probes bearing the nitroxide near the polar head group, a less restricted motion at low ionic strength was seen than with those labels with a nitroxide deeper within the hydrophobic tail of the membrane. Although these data clearly show an influence of ionic strength on membrane structure, and possibly a modified protein-lipid interaction, they cannot be simply discussed in terms of an altered membrane fluidity. PMID- 3034350 TI - Use of a fluorescence assay to monitor the kinetics of fusion between erythrocyte ghosts, as induced by Sendai virus. AB - The kinetics of the fusion process between erythrocyte ghosts, as induced by Sendai virus, were readily revealed by a simple fluorescence procedure previously employed to characterize the fusion of viruses with biological membranes. The method relies on the relief of fluorescence selfquenching of the membrane inserted probe octadecyl Rhodamine B chloride (R18) as occurs when labeled membranes fuse with unlabeled counterparts. The kinetics of R18 insertion into ghost membranes, the non-exchangeable properties of the fluorophore and the kinetics, and some characteristics of Sendai virus-induced fusion of ghosts, are described. We propose that the experimental approach may be particularly advantageous to obtain insight into the efficiency and mechanism of a wide range of fusogens, capable of inducing fusion of erythrocyte membranes. PMID- 3034351 TI - Multiwell cap assay: a simple objective method for the assessment of leukocyte locomotion in vitro. AB - A simple method for evaluating leukocyte locomotion in vitro has been developed and validated for several chemoattractants. The multiwell cap assay (MWCA) comprises chambers constructed from readily available disposable plastics and is quickly assembled, permitting large experimental protocols. Leukocytes which have migrated through a micropore filter are recovered and counted electronically yielding a precise, objective result. Coefficients of variation are approximately 6%. PMID- 3034353 TI - Favism: impairment of proteolytic systems in red blood cells. AB - Red blood cells (RBC) from favic patients are characterized by (a) severe oxidative damage (contributed by autoxidation of divicine and isouramil, two pyrimidine aglycones present in fava beans) and (b) greatly increased calcium levels. In vitro, both autoxidation of divicine and calcium loading produced marked alterations of proteolytic systems in intact RBC. Specifically, autoxidizing divicine inactivated procalpain, the proenzyme species of calcium activated cytosolic neutral proteinase, or calpain. Inactivation was much greater with glucose-6-phosphate dehydrogenase (G6PD)-deficient RBC than with normal RBC. On the other hand, loading of normal and G6PD-deficient RBC with calcium resulted in conversion of procalpain to calpain and eventual autoproteolytic inactivation of calpain itself, and extensive release of acid endopeptidase activity from the membranes into the cytosol. Damaged RBC from favic patients had significantly lowered procalpain activity and an abnormal subcellular distribution of acid proteinase activity that was found mostly in the cytosol. When purified calpain was incubated with membranes from acetylphenylhydrazine (APH)-treated RBC, significant proteolysis was observed affecting mostly band 3 and hemoglobin chains, ie, the two proteins involved in the onset of aggregation of Heinz bodies. Moreover, exposure of intact RBC to 20 mmol/L APH induced depletion of procalpain activity for which the time course was inversely related to formation of Heinz bodies. These findings support the role of procalpain in protecting G6PD deficient RBC from oxidant-induced Heinz body formation and imply that exhaustion of the procalpain-calpain system is an important step in the mechanisms of RBC damage and destruction in favism. PMID- 3034352 TI - Plasma cyclic nucleotide levels in acute leukemia patients. AB - To verify the clinical usefulness of extracellular cyclic nucleotide determination as a tumor marker, plasma cyclic AMP (cAMP) and cyclic GMP (cGMP) levels were measured in 70 normal subjects and 173 acute leukemia patients studied in different stages of their disease. Mean plasma cAMP levels were similar in leukemic and normal subjects, although in 48 patients in the active stage of the disease, first diagnosis, or relapse, the cAMP values were below the normal range, and most of these patients failed to respond to chemotherapy. Plasma cGMP levels were markedly elevated in untreated patients, normalized in all patients who attained complete remission, and increased promptly to pretreatment values in patients who relapsed, suggesting that their determination may be useful to monitor the patients' response to treatment. PMID- 3034354 TI - Aetiological factors in hepatocellular cancer. AB - The major risk factors for HCC are outlined in Table 2. Each factor may contribute to the multistep process of hepatocarcinogenesis. Hepatitis B virus and aflatoxins are the principal aetiological candidates and may be considered as initiators of the malignant state (see Figure 1). The majority of HCC arises via the cirrhotic pathway; the associated changes in the hormonal milieu may alter the handling of environmental carcinogens and the rounds of cell proliferation may increase sensitivity to sub-threshold doses of carcinogens. Exogenous androgens and oestrogens may operate through a similar mechanism to promote clonal expansion of preneoplastic cells. The importance of identifying the major aetiological factors lies in the possibility of prevention. The prognosis of HCC is dismal and it represents a major scourge in developing Third World countries. It is encouraging to think that the majority of cases could be prevented by the widespread use of hepatitis B vaccines and the development of intervention programmes against aflatoxin contamination of foodstuffs. PMID- 3034355 TI - Use of monoclonal antibodies for diagnosis and treatment of liver tumours. AB - Antibody-guided diagnosis can provide information regarding malignant disease which is not obtainable by conventional techniques and in some instances this approach can help to achieve prolonged tumour-free survival. Nevertheless, there are many technology improvements that need to be made if this method is to be widely applied for routine diagnosis and therapy. Already, encouraging therapeutic responses have been reported using 131I-labelled polyclonal and monoclonal antibodies in the treatment of primary and metastatic liver tumours. It is essential, however, to improve the radiolabelling of monoclonal antibodies for therapeutic use. It would be more convenient and effective to use radioactive isotopes which are pure beta emitters, e.g. 90Y and 32P which would allow for outpatient therapy. It is essential to evaluate the use of multiple antibodies or antibodies that bind to multiple antigenic targets within tumours to improve dose rate and total dose amplification. Finally, encouraging clinical pilot studies should be followed up by documented randomized clinical trials in order to define properly the clinical place of monoclonal antibodies, in both the diagnosis and the therapy of malignant disease, including primary and metastatic liver tumours. PMID- 3034356 TI - Surgery of liver tumours. AB - The liver is a segmental organ that allows resection through anatomically defined planes. The surgical management of an intrahepatic lesion, discovered either during investigation of hepatological symptoms or coincidentally, must involve an approach to investigation that carries a minimum risk and does not compromise subsequent excision of the lesion. Biopsy of an intrahepatic lesion found at laparotomy is essential, but attempts at early tissue diagnosis by percutaneous biopsy of operable tumours may lead to unnecessary morbidity and tumour spread. Preoperative studies often allow a firm pathological diagnosis to be made and ultrasonography, CT scanning and arteriography can be used to fully assess operability. Hepatocellular carcinoma (HCC) is the commonest primary liver cancer and is often found in association with cirrhosis and in patients with inadequate functional hepatic reserve. Surgical excision represents the only hope of cure for these patients and a 35% 5-year survival can be achieved by resection in the non-cirrhotic patient. Fibrolamellar HCC is less often associated with cirrhosis and is more often resectable with a better prognosis. Secondary tumours are often diffuse but about 5% of colorectal metastases are either solitary or confined to a resectable area of the liver. These tumours and secondary deposits from gastrointestinal endocrine tumours represent a small group of patients with potentially curable metastatic disease. Morbidity and mortality of operation depends on the extent of resection and the functional reserve of the liver. Local resections and resection for benign disease should carry no operative mortality. Major hepatic resection has a mortality of 3-5% and resection involving the structures at the hilus of the liver has an operative mortality of 10-12%. Liver transplantation in the management of neoplastic disease in the liver has yet to show any benefit over resectional surgery except where tumours have been discovered incidentally in the removed liver after transplantation for cirrhosis. PMID- 3034357 TI - Chemotherapy and radiotherapy of malignant hepatic tumours. AB - Cure of primary liver tumours remains possible only by surgery and early diagnosis will therefore continue to be important; the value of regular screening of cirrhotic patients for development of HCC by ultrasound scanning and estimation of AFP is now established. Prognosis of irresectable HCC depends largely on the general condition of the patient at the time of diagnosis and is better in the absence of cirrhosis. Radiotherapy has little role in the management of patients with HCC, but benefit with acceptable morbidity may be obtained from parenteral chemotherapy, with doxorubicin or its derivatives used as single agents, or with a combination of 5-FU and methyl-CCNU. There may be advantage from regional therapy given via the hepatic artery and early results from the combination of embolization with arterial doxorubicin are encouraging. The use of radiolabelled antibodies to tumour-related determinants of hormonal manipulation show promise. Worthwhile results from the non-surgical management of peripheral (intrahepatic) cholangiocarcinoma and primary hepatic sarcoma remain scarce. Isolated hepatic metastases from colorectal primaries may be resectable; for those that are not, results from regional chemotherapy with 5-FU or FUDR are encouraging, but cost and high morbidity currently limit more general application. PMID- 3034358 TI - The clinical features and natural history of malignant liver tumours. AB - As a broad generalization, there appears to be little intrinsic difference in the biological behaviour of the common malignant liver tumours in respect of presentation, clinical course, clinical features and prognosis. Whatever the tumour's origin, patients present with some combination of abdominal pain, hepatomegaly, weight-loss and general malaise and death occurs within 3 years of the onset of symptoms. It is the state of the non-tumorous liver (cirrhotic/non cirrhotic) and the anatomical site of the tumour (as with hilar cholangiocarcinomas) that are responsible for any significant differences. Metastatic carcinoid tumours, epithelioid haemangioendotheliomas, stage IV-S neuroblastomas and the fibrolamellar variant of HCC are exceptions to this rule with a genuinely better prognosis. PMID- 3034359 TI - Liver transplantation in the treatment of hepatic malignancy. PMID- 3034360 TI - Carcinoid and neuroendocrine tumours of the liver. PMID- 3034361 TI - Imaging techniques. PMID- 3034363 TI - [A case report on death due to an asthmatic attack observed in a guinea pig with TDI-induced asthma]. PMID- 3034362 TI - Immunohistochemical studies on small cell lung cancer. PMID- 3034364 TI - Growth of bacteria in an oil shale retort water by indigenous microorganisms. PMID- 3034366 TI - Spinal opiates. PMID- 3034365 TI - Receptors and functional linkage in membrane permeability: a quantum mechanical model. PMID- 3034367 TI - The effect of platelet activating factor on pulmonary beta-adrenoceptors. AB - An intravenous infusion of platelet activating factor (Paf) in the guinea-pig elicits an increase in bronchial responsiveness to the spasmogens, histamine and bombesin. Airways obstruction induced by bombesin in Paf-treated animals is poorly reversed by isoprenaline compared to comparable airways obstruction induced by bombesin in vehicle-treated animals. Isoprenaline induced a comparable dose-related relaxation in vitro of tracheal smooth muscle isolated from Paf- and vehicle-treated animals. No change in beta-adrenoceptor numbers or binding affinity was observed in lungs removed from Paf-treated animals in comparison with those from vehicle-treated animals, or after direct incubation with Paf in vitro. The reduced bronchodilator responsiveness to isoprenaline in Paf-treated animals is not related to changes in pulmonary beta-adrenoceptor function. These results suggest that non-spasmogenic elements may contribute to airways obstruction induced in hyper-responsive animals. PMID- 3034368 TI - Stress induced ACTH release in capsaicin treated rats. AB - The plasma concentrations of adrenocorticotropic hormone (ACTH) in rats pretreated with capsaicin as neonates were compared with those of control rats pretreated with the capsaicin vehicle. Capsaicin pretreatment has been shown earlier to abolish the increase in plasma ACTH concentration induced by cold stress while not affecting that induced by restraint stress. In the present experiments rats pretreated with capsaicin showed the same increase in plasma ACTH concentration in response to an i.v. infusion of ovine-corticotropin releasing factor as control rats pretreated with the capsaicin vehicle. Intraperitoneal injection of formalin, surgical stress and intravenous infusion of (-)-isoprenaline increased plasma ACTH concentrations in control rats. In capsaicin pretreated rats the increase in plasma ACTH was significantly attenuated. It is concluded that capsaicin-sensitive sensory neurones mediate the activation of pituitary ACTH secretion in response to somatosensory stimuli. The function of the corticotroph cells of the anterior pituitary is not impaired by capsaicin treatment. PMID- 3034370 TI - The prevalence of antibodies to an Epstein-Barr virus-induced polypeptide (EBNA 2) in the sera of rheumatoid arthritic families. AB - Using the protein immunoblot technique, antibodies to an Epstein-Barr virus induced 92 kD polypeptide (EBNA-2) were more frequently present in the sera of patients with rheumatoid arthritis and their consanguineous relatives when compared with a control group. No association of anti-EBNA-2 antibody with the HLA-DR antigens was observed. PMID- 3034371 TI - A comparison of clonogenic and radionuclide uptake assays for determining the radiation response of human small-cell lung cancer xenografts and cell lines. AB - Previous work has suggested that a radionuclide (3HTdR) uptake assay can provide a measure of drug and radiation sensitivity comparable with that given by the clonogenic assay. We have now extended this work to examine the radiation response of human small-cell lung cancer xenografts and cell lines with a wide range of plating efficiencies. Preliminary experiments using cultured cells indicated that irradiation of a single-cell suspension following disaggregation generally produced similar response data to those obtained when cultures were irradiated in situ and subsequently disaggregated. The response of eight cell lines to a single dose of 2 Gy was measured using both radionuclide uptake and clonogenic assays. There was no correlation between the two sets of data. Agreement between the radiation-response curves obtained using 3HTdR uptake and clonogenic assays was better for high plating efficiency xenografts (NCI-H69, COR L51) than for low plating efficiency xenografts (COR-L24, COR-L31). Radionuclide uptake indicated very shallow response curves for the low plating efficiency lines. Altering the time of radionuclide addition from the standard Day 4 to other times between Day 2 and Day 6 did not greatly change the indications of radioresponsiveness provided by the assay. Growth-curve experiments for line COR L24 showed that cell numbers at Day 4 after irradiation with 1.5 Gy or 3 Gy were very similar to those in unirradiated cultures. For NCI-H69, however, the cell numbers were very different for the different radiation doses. It appears that a high proportion of cells in lines such as COR-L24 take many days to show radiation damage as measured by reduced 3HTdR uptake. PMID- 3034369 TI - Antagonism of vasoconstriction induced by platelet-activating factor in guinea pig perfused hearts by selective platelet-activating factor receptor antagonists. AB - Platelet-activating factor (Paf) is a potent coronary vasoconstrictor in rat, guinea-pig, dog and pig. The present study investigated the mechanism and duration of Paf in guinea-pig isolated, Krebs-perfused hearts. Dose-related and sustained decreases in cardiac contractility and increases in coronary perfusion pressure were elicited by bolus doses of Paf (0.3-100 pmol). Platelet-activating factor (30 pmol) induced increases in the production of immunoreactive thromboxane B2 (TXB2), leukotriene B4 (LTB4) and LTC4, but not 6-keto prostaglandin F1 alpha (6-keto-PGF1 alpha). In addition, the release of leukotriene-like material following Paf was observed using on-line superfusion bioassay. The coronary vasoconstrictor actions of Paf were partially antagonized by the leukotriene receptor antagonist, FPL 55712 (1.9 microM), or by indomethacin (2.8 microM). The combined use of these compounds did not result in further significant inhibition. The Paf receptor antagonists, BN 52021 (30 microM) and L 652, 731 (10 microM), antagonized both the increase in coronary perfusion pressure and the decrease in cardiac contractility induced by Paf (10 100 pmol) in a surmountable and relatively selective manner. The effects of a bolus dose of 100 pmol Paf were sustained in excess of 18 min. Exogenous Paf underwent little metabolism on passing through the coronary circulation with only 2% being converted to lyso-Paf and approximately 4% being retained by the heart after 18 min of perfusion. These results suggest that the coronary vasoconstrictor actions of Paf are partially dependent on the release of vasoactive arachidonic acid metabolites. The extraordinary potency and the long lasting action of Paf indicate a potential role for this pro-inflammatory mediator in disorders of the coronary circulation. PMID- 3034372 TI - Effects of intradermal bradykinin after inhibition of angiotensin converting enzyme. AB - Inhibitors of angiotensin converting enzyme may cause angio-oedema. To see if this might be due to potentiation of the tissue effects of bradykinin the thickness of weals raised by intradermal injection of saline or 1, 3, or 10 micrograms bradykinin was measured before and three times after single doses of captopril, enalapril, or placebo. The mean thickness increased with increasing doses of bradykinin. It did not change with time after the administration of placebo or captopril but increased from 0.61 mm before enalapril to 1.12 mm two and a half hours and 1.06 mm five hours after enalapril was given. Five subjects flushed when given bradykinin after captopril and four after enalapril, but none flushed when given bradykinin after placebo. It is concluded that angiotensin converting enzyme inhibitors potentiate the effects of intradermal bradykinin in vivo and that this may partially explain why they cause angio-oedema in susceptible patients. PMID- 3034373 TI - Forced diuresis to reduce nephrotoxicity of streptozotocin in the treatment of advanced metastatic insulinoma. PMID- 3034374 TI - Induction of muscarinic acetylcholine, serotonin and substance P receptors in Xenopus oocytes injected with mRNA prepared from the small intestine of rats. AB - Serotonin and muscarinic acetylcholine (ACh) receptors were clearly induced in Xenopus oocyte injected with mRNA prepared from the small intestines of rats. Their response to ACh and serotonin was composed of 4 distinct components: fast and slow depolarization, slow hyperpolarization and large membrane potential fluctuation. About three-quarters of the injected oocytes responded to substance P. The response of the injected oocytes to substance P was transient and decayed even in the presence of substance P, indicating the presence of desensitization. However, the injected oocytes showed no response to 6 other drugs analyzed: adrenaline, noradrenaline, dopamine, gamma-aminobutyric acid, glycine and glutamate. PMID- 3034376 TI - Physiological studies of brainstem reticular connectivity. II. Responses of mPRF neurons to stimulation of mesencephalic and contralateral pontine reticular formation. AB - The connectivity between medial pontine reticular formation (mPRF) and the contralateral mPRF and between mPRF and the mesencephalic reticular formation (MRF) was studied by intracellular recordings of mPRF neuronal responses to microstimulation of the contralateral gigantocellular field (cFTG) portion of mPRF and ipsilateral MRF in unanesthetized, undrugged cats. There was a very high percentage (75-86%) of monosynaptic latency postsynaptic potentials (PSPs) in mPRF neurons in response to microstimulation of cFTG and MRF, and most PSPs (72 82%) were excitatory ones (EPSPs). The initial EPSPs from cFTG stimulation were characterized by a rapid rise time and a relatively constant latency, while those from MRF had a less rapid rise time and a longer plateau; EPSPs from both sites frequently led to spike potential generation. In contrast, the percentage of initial monosynaptic inhibitory PSPs (EPSPs) was less than 4% from each of these regions, statistically significantly less than that from bulbar FTM and bulbar FTG stimulation (about 12%) reported in the companion paper. Injection of depolarizing current in mPRF neurons unmasked hyperpolarizing PSP responses to stimulation that followed initial depolarizing PSPs. Intracellular HRP labeling indicated that these data were from recordings from neurons with 20-100 microns diameters, with 80% greater than 40 microns. Neurons with a different discharge pattern for this area of the pons, a stereotyped burst pattern, were recorded just ventral to mPRF; this discharge pattern resembled that found in inhibitory interneurons in other central nervous system regions. There were no differences in the density and pattern of orthodromic PSPs between those mPRF neurons that were antidromically activated from cFTG and the general population that was not antidromically activated from cFTG or other stimulated sites; this suggests, when combined with data of the companion paper, an identity of input and output elements in mPRF with respect to synaptic response properties. The high degree of connectivity between reticular regions may furnish a substrate for functional interaction. PMID- 3034375 TI - After-hyperpolarizations produced in frog motoneurons by excitatory amino acid analogues. AB - After-hyperpolarizations (AHPs) produced in frog motoneurons by applications of the excitatory amino acid analogues quisqualate (QUIS), N-methyl-D-aspartate (NMDA), and kainate (KA) were studied in the isolated hemisected frog spinal cord using sucrose gap techniques. AHPs were present following 98% of QUIS-induced depolarizations, but were seen in only 35% and 15% of NMDA- and KA-evoked responses respectively. AHPs produced by QUIS are produced both by direct effects of QUIS on motoneuron membranes and by indirect effects mediated through a synaptic process involving interneurons. Thus, application of Mg2+, Mn2+, or tetrodotoxin (TTX) in concentrations sufficient to block synaptic transmission and interneuronal firing, reduced, but did not abolish the AHPs produced by QUIS. In contrast, NMDA- and KA-AHPs appear to be entirely mediated by indirect means as block of synaptic transmission and interneuronal firing eliminated AHPs produced by these substances. Exposure of the cord to Mn2+ after addition of TTX did not affect the size of QUIS-AHPs. In the presence of TTX, QUIS-AHPs were reduced or completely blocked by addition of dinitrophenol (DNP) and sodium cyanide, by dihydro-ouabain, by removal of K+ from the superfusate, by cooling, and by replacement of 50% of the external Na+ with Li+. The results suggest that the QUIS-AHPs are largely the result of the direct effect of the excitatory amino acid agonist on motoneuron membranes and is caused by activation of an electrogenic Na+ pump. AHPs following depolarizations evoked by NMDA and KA are presumably the result of indirect actions of these latter analogues on interneurons. PMID- 3034377 TI - Effects of stimulating the subcoeruleus-parabrachial region on the non-noxious and noxious responses of T1-T5 spinothalamic tract neurons in the primate. AB - The effects of electrical stimulation of the subcoeruleus-parabrachial (SC-PB) region on the discharge rate of upper thoracic spinothalamic tract (STT) neurons were investigated in 21 monkeys anesthetized with alpha-chloralose. STT cells were antidromically activated from the medial thalamus (MT) and the ventral posterior lateral nucleus (VPL) and received viscerosomatic convergent input from the cardiopulmonary sympathetic afferents and the left chest-forearm region. Stimulation of the SC-PB region inhibited the activity of all 30 STT neurons studied in the T1-T5 regions of the spinal cord. The minimum average current required to decrease the discharge rate of 22 cells exhibiting spontaneous activity was 89 +/- 10 microA (100 Hz, 100 microseconds duration). Currents as high as 300 microA completely inhibited the activity of most cells. Examination of the importance of frequency of stimulation from the SC-PB area on 8 cells revealed that impulses of at least 40 Hz (208 +/- 37 microA, 100 microseconds duration) were necessary to inhibit the spontaneous activity by 60%. Higher frequencies produced greater degrees of inhibition. Stimulation of the SC-PB region also inhibited the response of 23 of 23 neurons excited by noxious pinch and 11 of 11 wide dynamic range cells stimulated by innocuous input such as blowing or brushing hair. No differences in the inhibition produced by SC-PB stimulation on cells projecting to VPL (n = 20), MT (n = 5), or both VPL and MT (n = 5) were observed. These results demonstrate that the SC-PB region may be an important brainstem site for descending inhibition of both noxious and innocuous somatic input to upper thoracic STT cells in the primate. PMID- 3034378 TI - Physiological studies of brainstem reticular connectivity. I. Responses of mPRF neurons to stimulation of bulbar reticular formation. AB - The connectivity between medial pontine reticular formation (mPRF) and bulbar reticular formation (BRF) was studied by intracellular recordings of mPRF neuronal responses to microstimulation of BRF in unanesthetized, undrugged cats. There was a very high percentage (75-90%) of monosynaptic latency postsynaptic potentials (PSPs) in mPRF neurons in response to microstimulation of 3 BRF areas: the magnocellular tegmental field (FTM), the bulbar gigantocellular tegmental field (BFTG), and bulbar lateral tegmental field (BFTL). The type of initial orthodromic response produced in mPRF neurons by BRF stimulation was predominantly (75-95%) a monosynaptic excitatory PSP (EPSP) which was characterized by a rapid rise time, a nearly constant latency, and often led to spike potential generation. In contrast, the percentage of initial monosynaptic inhibitory PSPs (IPSPs) was much lower for FTM (12.3%), for BFTG (12.5%) and was zero for BFTL. While microstimulation techniques alone cannot differentiate between excitation of fibers of passage and neuronal somata, the very high percentage of initial EPSPs in our data and the anatomical evidence for dense BRF to mPRF neuronal projections as compared with less dense projections from fibers passing through BRF to mPRF suggest that excitatory BRF-mPRF connections are predominant. The high degree of connectivity between BRF and mPRF may furnish an important substrate for functional interaction. Comparison of the mPRF neuronal population that was not antidromically activated by FTM microstimulation vs the mPRF neuronal population that was antidromically activated from FTM and also studied for orthodromic responsiveness showed no statistically significant differences between these populations on the parameters of percentage of monosynaptic input, monosynaptic initial EPSPs, monosynaptic initial IPSPs and presence of a PSP with a latency of less than 5 ms. For BRF connectivity this suggests an identity of mPRF input and output neurons with respect to synaptic response properties. PMID- 3034379 TI - Relation of olfactory bulb and cortex. II. Model for driving of cortex by bulb. AB - The major projection pathway of the olfactory bulb is by way of the lateral olfactory tract (LOT) to the olfactory cortex. Oscillatory bursts of extracellular potential appear during inspiration in both bulb and cortex. Based on anatomical and physiological considerations, a model was proposed, consisting of a bulbar transmitter, a conduction line representing axons in the LOT, and a cortical receiver. The model predicted the relation between phase and frequency of bulbar and cortical burst pairs, based on the expectation that the bulb drives the cortex. Experimental phase-frequency plots were computed from bursts of 9 bulbocortical electrode site pairs from each of 10 rabbits. For each site pair, the model predicted the expected range of the joint variation of phase and frequency, using the known distance between the bulbar and cortical sites. The model was highly successful (greater than 95% prediction accuracy) for one quarter of the total number of site pairs examined. The wide range of variation for the rest of the data suggested that higher order interactions are responsible for the phase relation between bulb and cortex. Convergence of input, independence of the cortical generator, cortical feedback to the bulb and synchronization by an outside source are all discussed as possible contributors to this variation. PMID- 3034380 TI - Autoradiographic visualization and characterization of [3H]ouabain binding to the Na+,K+-ATPase of rat brain and pineal. AB - Ouabain binds to the catalytic subunit of Na+,K+-ATPase and specific [3H]ouabain binding can be used as a measure of the number of active enzyme molecules present in a given tissue. Specific [3H]ouabain binding can be demonstrated in frozen, cryostat sections from rat brain and pineal and these sites have the characteristics of Na+,K+-ATPase. Incubations carried out in the absence of ATP or the presence of excess unlabeled ouabain reduces specific binding by greater than or equal to 98%. The addition of K+ or omission of Mg2+ also result in a decrease in specific binding. Strophanthidin, digoxin and digoxigenin displace [3H]ouabain binding with IC50 values of 0.73, 0.48 and 1.4 microM, respectively. Scatchard analyses of specific [3H]ouabain binding in brain sections shows a single class of non-interacting binding sites with an apparent affinity (Kd) of 339 nM and a maximal binding capacity (Bmax) of 34.9 pmol/mg protein. [3H]Ouabain binding is unevenly distributed throughout the brain with the olfactory nuclei, superior colliculus, dentate gyrus, pontine nuclei and pineal gland having a relatively high density of binding sites. The outer layers (1-3) of the cerebral cortex show more labeling than the inner layers (4-6) and most other brain areas have intermediate levels of [3H]ouabain binding sites, whereas white matter has virtually no specific binding. Computer-assisted densitometry was used to measure changes in specific [3H]ouabain binding after kainic acid injection into the caudate nucleus. An initial increase in [3H]ouabain binding was observed at 1 and 24 h after lesioning and a decrease in [3H]ouabain binding was evident by 9 days after lesioning.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3034383 TI - Kynurenic acid blocks suprachiasmatic nucleus responses to optic nerve stimulation. AB - An in vitro slice preparation of the mouse hypothalamus was used to determine the effects of pharmacological agents on the field potentials that are evoked in the suprachiasmatic nucleus (SCN) by stimulation of the optic nerve. Postsynaptic components of these responses were identified by lowering the concentration of calcium in the superfusate. Bath application of kynurenate, an antagonist of excitatory amino acid neurotransmission, reversibly blocked postsynaptic responses in the SCN. The evoked responses in the SCN were not affected by the acetylcholinergic agents (+)-tubocurarine, scopolamine, physostigmine, or carbachol. These results suggest that excitatory amino acid receptors mediate responses of SCN neurons to retinal input, but do not support a role for acetylcholine. PMID- 3034381 TI - Light absorbance changes in the mouse carotid body during hypoxia and cyanide poisoning. AB - The isolated carotid body (CB) of the mouse was bathed with modified Tyrode's solution (pO2, 170 Torr) and transilluminated at different wave-lengths (500-620 nm). Difference spectra (normoxia vs hypoxia--pO2, 10 Torr) showed peaks of increased light transmittance (LT) at 500 and especially at 540 and 580 nm. There were troughs of transmittance decrease at 560 and 600 nm. Similar spectra (normoxia vs NaCN 10 mM) revealed increased LT at 500 and 590 nm, whereas there was decreased LT at 550 nm. Therefore, absorbance changes induced by hypoxia and cyanide were different. As as control for residual hemoglobin (Hb) in the tissues, a segment of carotid artery was filled with blood and the same light scanning was performed. The differential signals evoked by hypoxia were much larger. There was some LT increase at 500 and 540 nm, but this effect was especially marked at 570 and 590 nm. NaCN showed the same absorbance pattern, but peaks were of lower amplitude. Thus, the CB-produced signals were not elicited by residual Hb. A stepwise decrease in the medium pO2 (from 170 to 10 Torr) surrounding the CB produced a progressive increase in LT at 540 and 580 nm starting at 120 Torr. A different pattern was evoked by 550 (cytochrome c) and 600 (cytochrome aa3) nm. With 550 nm, transmittance began to increase at about 70 Torr. LT decreased with 600 nm starting at about 120 Torr.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3034382 TI - Regulation of central and peripheral benzodiazepine receptors in progesterone treated rats. AB - The effects of progesterone treatment to male rats on cortical and hippocampal central benzodiazepine (BZ) receptors and peripheral BZ binding sites (PBS) in kidney, testis and heart were evaluated. Chronic progesterone treatment resulted in a 30% elevation of the density of central BZ receptors in cerebral cortex accompanied by a 24% augmentation of PBS in the kidney. No significant change in the density of central BZ receptors in the hippocampus or of PBS in the testis or heart was observed. No change in the affinity of [3H]flunitrazepam and [3H]PK 11195 binding to central BZ receptors and PBS, respectively, was observed. The increase in PBS in the kidney might be associated with the natriuretic effect of this hormone and the adaptatory increase in aldosterone secretion. The up regulatory effect of progesterone on cortical central BZ receptors may be involved in the neurobehavioral activities of progesterone. PMID- 3034384 TI - Protein phosphorylation in primary astrocyte cultures treated with and without dibutyryl cyclic AMP. AB - Protein phosphorylation was investigated in primary rat astrocyte cultures treated with and without dibutyryl cyclic AMP. Astrocytes maintained in dibutyryl cyclic AMP for several weeks displayed increased phosphate incorporation in 5 protein bands (55, 52, 45, 43 and 28 kDa) while incorporation in one band (42 kDa) was decreased. Phosphate incorporation in several other protein bands was unchanged. Calcium-dependent phosphate incorporation was also altered by prior exposure of the cells to dibutyryl cyclic AMP: addition of calcium to broken cell preparations resulted in increased incorporation in 75, 53 and 52 kDa while decreased incorporation occurred in 100 kDa. These differences in protein phosphorylation may be related to the previously reported biochemical and morphological changes brought about by dibutyryl cyclic AMP and may provide insights into the mechanisms of reactive gliosis. PMID- 3034385 TI - [The importance of ultrasound in the early diagnosis of invasive forms of trophoblastic disease]. PMID- 3034386 TI - Response of osteoblastic clonal cell line (MC3T3-E1) to [Asu]eel calcitonin at a specific cell density or differentiation stage. AB - Clone MC3T3-E1 cells isolated from newborn mouse calvaria is an osteogenic cell line which retains an ability to differentiate into osteoblastic cell in vitro. The effect of [Asu]eel calcitonin (ECT) on clonal MC3T3-E1 cells was investigated at different stages of differentiation. ECT caused an increase in alkaline phosphatase (ALP) activity. The stimulative effect was demonstrated to be dependent upon cell density or differentiation stage. At a cell density of 1.18 X 10(5)/cm2 cells were incubated with ECT for 2 days. The treatment by ECT caused an increase in ALP activity. A specific response to ECT dependent on the cell density was observed in a narrow range of cell density. Moreover this range of cell density responsible to ECT was found to be a rapid differentiation stage of MC3T3-E1 cells. These results suggest that calcitonin stimulates differentiation of osteoblast. In addition to these results, cellular adenosine-3',5'-cyclic monophosphate (cAMP) level was raised by ECT treatment at a cell density of about 1.4 X 10(5) cell/cm2 and this response was also specific for cell density. At cell density lower or higher than this density no stimulative effect by ECT was observed. On the other hand, N6,O2-dibutyryl adenosine-3',5'-cyclic monophosphate (db-cAMP) and theophylline caused an increase in ALP activity in wide cell density range. These results indicate that an increase in ALP activity by ECT is mediated by intracellular cAMP and that the specific response to ECT dependent on the cell density is regulated in the process of cAMP formation and/or in the preceding process of cAMP formation. PMID- 3034388 TI - Nonamelogenin components of porcine enamel in the protein fraction free from the enamel crystals. AB - The enamel protein fraction free from enamel crystals was investigated to determine the existance of nonamelogenin like enamelin. Enamel proteins were extracted by neutral and alkaline buffers from the porcine immature enamel at an early stage of development and resolved into four fractions on Sephadex G-100 gel filtration in a carbonate buffer (pH 10.8). The first eluted fraction contained the aggregate of proteins from 13,000 daltons to 142,000 daltons in molecular size and most of these proteins were found to differ from amelogenin by their different solubility against 25% isopropanol on acrylamide gel, and their amino acid composition. These nonamelogenins showed the property of closely associating with synthetic hydroxyapatite under dissociative conditions, and their electrophoretic properties and amino acid compositions were quite similar to those of the enamelins prepared from porcine immature enamel. PMID- 3034387 TI - Role of carbonic anhydrase in bone resorption: effect of acetazolamide on basal and parathyroid hormone-induced bone metabolism. AB - The effects of the carbonic anhydrase inhibitor acetazolamide on basal and parathyroid hormone (PTH)-induced bone metabolism were studied to evaluate the manner in which acetazolamide inhibits bone resorption. Half-calvaria from 5 to 6 day-old mice were cultured using the following treatments: control; acetazolamide (10, 33, or 100 microM); PTH (16.7 nM bovine PTH 1-34); acetazolamide + PTH. The effects of acetazolamide on PTH-induced cAMP accumulation and protein synthesis were determined. Media from bones cultured for 48 hours were analyzed for calcium to assess bone resorption, glucose to assess calvarial glucose utilization, and lactic acid to assess calvarial lactic acid release. Media were also assayed for beta-glucuronidase activity as an indicator of lysosomal enzyme release and for lactate dehydrogenase activity as an indicator of cytosolic enzyme release and cytotoxicity. Acetazolamide at 100 microM completely inhibited PTH-induced bone resorption. This inhibition did not appear to be due to cell death, as acetazolamide did not increase lactate dehydrogenase release. Acetazolamide had no effect on PTH-enhanced cAMP levels, indicating that receptor binding and adenylate cyclase activation were unaffected. Acetazolamide alone did not alter calvarial protein synthesis, but did significantly inhibit protein synthesis in the presence of PTH. PTH significantly enhanced calvarial glucose utilization, lactic acid release, and beta-glucuronidase release. Acetazolamide inhibited all of these PTH-induced parameters in a manner that roughly paralleled its inhibition of bone resorption; acetazolamide alone had no effect on the basal values. Our results indicate that acetazolamide inhibition of bone resorption in vitro may involve general alterations in hormonally stimulated bone cell metabolism secondary to carbonic anhydrase inhibition. PMID- 3034389 TI - Relationship of urotensin I induced vasodilatory action in rat thoracic aorta to Ca2+ regulation. AB - The relaxant effects of the synthetic fish neuropeptide urotensin I were examined in helical strips of rat aorta. In K+-depolarized aorta strips, urotensin I and verapamil competitively inhibited Ca2+-induced contractions. Urotensin I relaxed, in a concentration-dependent manner, the contraction produced by the Ca2+ ionophore A23187, whereas verapamil had no effect on this contraction, even at a concentration of 10(-5) M. In the absence and presence of extracellular Ca2+, urotensin I inhibited both components of the contractions elicited by norepinephrine or urotensin II, another fish neuropeptide. Verapamil reduced only the norepinephrine or urotensin II induced contraction in the presence of extracellular Ca2+, with little or no change in the contraction in Ca2+-free buffer. The urotensin I induced relaxation response in aortic strips contracted by 40 mM KCl was enhanced by pretreatment with papaverine or forskolin. Pretreatment with dibutyryl cAMP did not significantly alter the action of urotensin I. The presence or absence of endothelial cells did not change the response to urotensin I. These results suggest that urotensin I antagonizes the action and (or) mobilization of extracellular and intracellular Ca2+. PMID- 3034391 TI - Effect of ethanol on the glucose-induced movements of protons across the plasma membrane of Saccharomyces cerevisiae NCYC 431. AB - The study of glucose-induced proton fluxes in Saccharomyces cerevisiae NCYC 431 showed a decrease of proton net efflux by ethanol across the plasma membrane of energized cells. Furthermore a negative net proton efflux (an influx) occurred from a given ethanol concentration (between 1.3 and 1.5 M) whatever the experimental conditions used, thus allowing the definition of a nil-net exchange step where no net movement of protons across the plasma membrane could be observed. A new technique of ethanol tolerance determination in yeast based upon a correlation for the same ethanol concentration between both the collapse of the proton gradient and the growth cessation in cultures supplemented with ethanol after 8 h incubation was proposed. The defined method also showed a cumulated effect of temperature and ethanol on Saccharomyces cerevisiae NCYC 431. PMID- 3034390 TI - Barbiturate effects on hippocampal excitatory synaptic responses are selective and pathway specific. AB - Barbiturate actions on excitatory synaptic responses in CA 1 and dentate regions of hippocampal slices were studied to determine whether different effects occur on anatomically distinct synaptic pathways. Pentobarbital facilitated transmission between stratum radiatum inputs and CA 1 neurons at low concentrations (0.02-0.08 mM) and produced postsynaptic depression at higher concentrations. Only depression was observed for stratum oriens inputs to CA 1 and perforant path inputs to dentate granulae neurons. The (+) isomer of pentobarbital was approximately four times more potent than the (-) isomer of racemic mixture. Phenobarbital (0.04-0.12 mM) produced only depression of synaptic responses in CA 1 and dentate pathways. Comparison of effect on field excitatory postsynaptic potentials and population spike responses indicated that the barbiturates act at selective and pathway-specific sites. The results provide further evidence for specific cellular and membrane recognition sites for barbiturate action. PMID- 3034392 TI - Sick at sea: outbreaks prompt reinstatement of cruise ship inspections. PMID- 3034393 TI - Nutrition in the prevention of neoplastic disease in the elderly. AB - This review focuses on specific effects of diet on cancer risk and the relevance of these dietary effects to the risk of cancer in the elderly. The authors address the impact of certain dietary factors on cancer risk by reviewing their roles in two distinct phases of carcinogenesis: "imitation" and "promotion." PMID- 3034394 TI - Testicular failure in patients with extragonadal germ cell tumors. AB - Eight patients with mediastinal or retroperitoneal germ cell tumors who had undergone testicular biopsy or orchiectomy were retrospectively analyzed for primary testicular abnormalities, subfertility, and abnormal sex hormone levels. Testicular tissue was abnormal in all patients, revealing peritubular fibrosis (six), decreased spermatogenesis (eight), interstitial edema (five), Sertoli cells only (one), and Leydig cell hyperplasia (two). Detailed hormone analysis in five patients revealed elevations of luteinizing hormone in four, decreased serum testosterone in two, elevations of estradiol in two, and elevation of human chorionic gonadotropin in one patient. A history of infertility was documented 2 months to 13 years before presentation in four patients and suspected in another. Extragonadal germ cell tumors, like their testicular counterparts are associated with primary germ cell defects, some of which seem to be independent of gonadotropin production by the tumor. In addition, the rather high incidence of antecedent infertility suggests that either a congenital or acquired primary germ cell defect contributes to defective spermatogenesis and the development of cancer in incompletely migrated germ cells. PMID- 3034395 TI - Salivary gland tumors of the tongue. Analysis of 55 new cases and review of the literature. AB - Fifty-five cases of primary salivary gland tumors of the tongue from the files of the Armed Forces Institute of Pathology are reported and analyzed and the results compared with the information in the literature. Five tumors were benign and 50 were malignant. The average age at presentation was 47 years for the benign tumors and 54.3 years for the malignant ones. Although the overall benign/malignant ratio was 1:10, women were more likely to have a malignant tumor than were men. The site of 80% of the benign tumors was the middle to anterior portion of the tongue, whereas over 85% of malignant tumors involved the base. Clinical signs and/or symptoms related to the site aroused suspicion in some cases but often were of short duration and in over 60% of cases did not occur. The most common benign tumor type was the myoepithelial variant of the benign mixed tumor. The most common malignant tumor type was the low-grade mucoepidermoid carcinoma (38%) followed by adenocarcinoma (20%), high-grade mucoepidermoid carcinoma (14%), adenoid cystic carcinoma (10%), and clear cell carcinoma (8%), with occasional basaloid, papillary cystadenocarcinoma, acinic cell and mucus-producing adenocarcinoma. Treatment was similar to that of other accessory salivary gland neoplasms of similar histologic type and clinical stage. Prognosis worsened with high histologic grade, old age, and advanced clinical disease at presentation. PMID- 3034396 TI - Translocation (3;8;8)(p22 or p23;p23;q12) in a case of pleomorphic adenoma: similarity to a primary cytogenetic abnormality detected in an endometrial adenocarcinoma. AB - Cytogenetic findings on the recurrence of a pleomorphic adenoma of the parotid gland are herein reported. The tumor showed an abnormal chromosome #8, which was very similar to a marker chromosome recently described as the sole abnormality in an endometrial adenocarcinoma. PMID- 3034397 TI - Cytogenetic and molecular characterization of a masked Philadelphia chromosome in chronic myelocytic leukemia. AB - A case of typical chronic myeloid leukemia with an apparently Philadelphia negative karyotype is described. Molecular studies confirmed the cytogenetic interpretation of a standard Ph rearrangement, with secondary involvement of 22q- in a translocation with chromosome #5, leading to its masking. The chromosomal regions engaged in the standard t(9;22) were not modified and the molecular rearrangements of Ph were also conserved. The hematologic and clinical features were apparently not influenced by the events leading to the masking of Ph. Further similar observations with both cytogenetic and molecular characterization are needed to better identify the possible clinical consequences of these complex changes. PMID- 3034398 TI - Chromosomal abnormalities in a primary small cell lung cancer. AB - A cytogenetic analysis of a fresh primary tumor specimen of small cell lung cancer showed a del(3)(p14p23) in the majority of metaphases. Additional clonal changes were found in the karyotype. No abnormalities for Ha-ras, Ki-ras, N-ras, myb, or myc were detected by Southern blot analysis of the tumor DNA. PMID- 3034400 TI - Inhibition of 12-O-tetradecanoylphorbol-13-acetate-enhanced transformation in vitro by radical scavengers. AB - The inhibitory effects of some scavengers of oxygen radicals were studied, using a two-stage transformation assay system in vitro 3-methylcholanthrene (3-MC) initiation and 12-O-tetradecanoylphorbol-13-acetate-(TPA)-promotion in Balb 3T3 cells. Mannitol, a scavenger for hydroxyl radicals, strongly inhibited the TPA enhanced transformation in a dose-dependent manner. Superoxide dismutase (SOD) and catalase, which are specific scavengers for O2-. and H2O2, respectively, also inhibited the transformation. These results suggest that oxygen radicals may play an important role in the TPA-enhanced transformation in Balb 3T3 cells. PMID- 3034399 TI - Cross-resistance to ouabain in a murine leukemia cell variant selected for cis dichlorodiammineplatinum(II) resistance. AB - A murine leukemic cell line (R1.1) variant (R1.1/CDDPR-E8) resistant to cis dichlorodiammineplatinum(II)(CDDP) was also found to be resistant to ouabain, a postulated specific inhibitor of sodium-potassium ATPase. The variant established by the culture of parental cells in step by step increasing concentrations of CDDP, exhibited 11-fold higher resistance to CDDP than the parental R1.1 cells. The present study suggests that a mutational change leading to an alteration in cell membrane characteristics associated with ouabain has also changed the sensitivity of cells against CDDP. Alternatively, the present data may indicate that the cytotoxicity of CDDP is closely linked to its effects on cell membrane. PMID- 3034401 TI - Expression of melanoma-associated antigens in rapidly dividing human melanocytes in culture. AB - Conditions were established to induce rapid clonal growth of melanocytes from newborn foreskin. Surface antigen expression was analyzed using monoclonal antibodies derived by immunization of mice with melanoma cell, melanocyte, and placental membrane preparations. Unlike resting melanocytes in normal skin, cultured melanocytes expressed most major melanoma-associated antigens tested, e.g., nerve growth factor receptor, proteoglycan, transferrin-related Mr 97,000 protein antigen, Mr 120,000 protein, and gangliosides 9-O-acetyl GD3 and GD3. HLA DR antigen and ganglioside GD2 were expressed at very low levels or not expressed. After several subpassages, most melanocyte cultures, including clones and melanocytes, initially sorted by rosetting with monoclonal antibody to nerve growth factor receptor, lost their characteristic bipolar morphology and expression of nerve growth factor receptor and Mr 97,000 antigen but continued to express high molecular weight proteins such as proteoglycan, Mr 130,000/105,000 and 120,000 antigen. The few melanocyte cultures that did maintain their characteristic bipolar to spindle morphology continued to express all melanoma associated antigens and even began to express HLA-DR antigens. Melanocytes cultured in the presence of the phorbol ester 12-O-tetradecanoylphorbol-13 acetate also maintained their bipolar morphology, were often pigmented, and continued to express melanoma-associated antigens for several passages; they did not express HLA-DR antigen. Our studies indicate that rapidly proliferating melanocytes in culture undergo antigenic changes associated with malignancy. PMID- 3034403 TI - 3'-Azido-3'-deoxythymidine in feline leukemia virus-infected cats: a model for therapy and prophylaxis of AIDS. AB - Due to similarities between human immunodeficiency virus and feline leukemia virus, the etiological agents of acquired immunodeficiency syndromes in humans and cats, the feline system was used as a model to conduct preliminary investigations as to the efficacy of the thymidine analogue 3'-azido-3' deoxythymidine (AZT) as a therapeutic and preventive agent against retroviruses. In vitro evaluations of AZT cytotoxicity and its antiviral effects were conducted. Subsequently, 50 6-week-old specific pathogen free kittens were inoculated with a highly immunosuppressive strain of Richard-Feline Leukemia Virus. These cats were randomly subdivided into smaller groups with initiation of AZT treatment at variable times postinfection. All animals were periodically monitored for circulating infectious virus particles and virus-neutralizing antibodies. Their clinical condition was closely followed throughout the 6-week AZT treatment phase and for several months thereafter. The results indicate that AZT prevents retrovirus infection if administered immediately following virus exposure, and may also reduce retrovirus replication if administered to previously infected animals. Some of the treated cats developed neutralizing antibodies against the virus and became resistant to subsequent viral challenge. Future trials with this drug, both for the prevention and treatment of retroviral diseases in humans and animals, are warranted. PMID- 3034404 TI - Activated c-Ha-ras oncogene with a guanine to thymine transversion at the twelfth codon in a human stomach cancer cell line. AB - The rat fibroblast cell line Rat 1 was transfected with total DNA of a gastrocarcinoma cell line, BGC-823. The transforming gene was cloned from the genomic library of the secondary transformants using in situ hybridization with a probe of the human Alu repeat sequence. This cloned gene is homologous to the protooncogene c-Ha-ras. The activation lesion of the transforming gene was identified by sequence analysis as a single nucleotide substitution of thymine for guanine in the 12th codon. This results in the substitution of valine for glycine at the 12th amino acid of the Mr 21,000 protein. PMID- 3034405 TI - Causal role for an activated N-ras oncogene in the induction of tumorigenicity acquired by a human cell line. AB - ras oncogenes have been found in approximately 15% of the human tumors analyzed. However, a causal role for these genes in the tumorigenesis of human cells has yet to be shown. Tumorigenic late-passage PA-1 human teratocarcinoma cells (E-PA 1) contain an activated N-ras gene. In this report evidence is presented that nontumorigenic early passage revertant PA-1 cells (E-PA-1) contain only the germ line protooncogene. Introduction by gene transfer of the activated L-PA-1 oncogene induces E-PA-1 cells to form tumors, suggesting that the activated N-ras oncogene has a causal role in the tumorigenesis of these cells. PMID- 3034402 TI - Effect of aldehyde dehydrogenase inhibitors on the ex vivo sensitivity of human multipotent and committed hematopoietic progenitor cells and malignant blood cells to oxazaphosphorines. AB - The ex vivo sensitivity of human multipotent and committed hematopoietic progenitor cells and several cultured human malignant blood cell lines to analogues of "activated" cyclophosphamide, namely, 4-hydroperoxycyclophosphamide and mafosfamide, and to phosphoramide mustard was quantified with and without concurrent exposure to an inhibitor of aldehyde dehydrogenase activity, namely, disulfiram, cyanamide, diethyldithiocarbamate, or ethylphenyl(2 formylethyl)phosphinate. Inhibitors of aldehyde dehydrogenase activity potentiated the cytotoxic action of 4-hydroperoxycyclophosphamide and mafosfamide toward all of the hematopoietic progenitors; they did not potentiate the cytotoxic action of phosphoramide mustard toward these cells. Potentiation of the cytotoxic action of mafosfamide toward cultured human malignant blood cells was minimal. Spectrophotometric assay revealed little NAD-linked aldehyde dehydrogenase activity present in the cultured human tumor cell lines as compared to that found in normal mouse liver or oxazaphosphorine-resistant L1210 cells. Cellular aldehyde dehydrogenases are known to catalyze the oxidation of 4 hydroxycyclophosphamide/aldophosphamide, the major intermediate in cyclophosphamide bioactivation, to the relatively nontoxic acid, carboxyphosphamide. Thus, our findings indicate that human multipotent hematopoietic progenitor cells contain the relevant aldehyde dehydrogenase activity, the relevant activity is retained upon differentiation to progenitors committed to the megakaryocytoid, granulocytoid/monocytoid, and erythroid lineages, and the relevant activity may be lost or diminished upon transformation of hematopoietic progenitors to malignant cells. PMID- 3034406 TI - Growth-related elevations of DNA topoisomerase II levels found in Dunning R3327 rat prostatic adenocarcinomas. AB - The relationship between DNA topoisomerase II expression and mammalian cell proliferation has been evaluated by determining enzyme levels in normal and neoplastic rat prostate tissues. By activity assay and by immunoblot analysis using anti-topoisomerase II antiserum, topoisomerase II levels were found to be elevated in both the Dunning R3327-H and the Dunning R3327-G rat prostatic adenocarcinomas over levels assayed in the normal rat dorsal prostate. Immunohistochemical studies using the antitopoisomerase II antiserum revealed that a greater fraction of nuclei contained detectable levels of topoisomerase II in tissue sections prepared from each of the Dunning tumors than in rat dorsal prostate tissue sections. The Dunning R3327-H and R3327-G tumors grow at different rates in vivo (J. T. Isaacs and D. S. Coffey, Clin. Oncol., 2: 479-498, 1983). When measured topoisomerase II levels were compared to known growth parameters for each of the tissues studied, topoisomerase II expression was found to be correlated with tissue growth rate. PMID- 3034408 TI - Amplification of IMR-32 clones 8, G21, and N-myc in human neuroblastoma xenografts. AB - Amplification of clones 8, G21, and N-myc, which were derived from human neuroblastoma cell lines IMR-32 and NB-19, were studied in nine neuroblastoma xenografts. N-myc was amplified from 50- to 120-fold in eight of nine xenografts, clone 8 was amplified in five of the xenografts, and clone G21 was amplified in four of these five. Each of these clones was localized by in situ hybridization to homogeneously staining regions in metaphase spreads of xenograft chromosomes. In one xenograft a DNA rearrangement of clone 8 was observed, and only two of the sequences detected by G21 were amplified. Restriction enzyme mapping indicated that the rearrangement within clone 8 occurred at a position close to the rearrangement previously noted in neuroblastoma cell line NB-9. PMID- 3034407 TI - Differential behavior of human bronchial carcinoma cells in culture. AB - A feeder layer culture system suited to grow carcinoma cells derived from solid human lung tumors was developed. This report deals with culturing of the four main histological types of lung carcinomas observed in 37 patients: 19 squamous cell, 6 adenocarcinomas, 7 small cell, and 5 large cell carcinomas. The cultures were initiated from 24 fresh human surgical specimens and from 14 human lung tumors grown as xenografts in nude mice. Three different patterns of behavior in culture were found to be characteristic for squamous cell, adenocarcinomas, and small cell carcinomas, respectively. The culture pattern presented by the primary cultures did not appreciably change after passaging in vitro for periods of up to 2 years, even after infinite cell lines were established. Cultures of large cell carcinoma showed one or more of these patterns. From these patterns cells could be cloned and subsequently cultured as separate stable lines. The system described facilitates the identification of specific types of human lung carcinomas almost immediately (within 1 h) after plating (Phase I) as well as during culture. PMID- 3034409 TI - Heterogeneous cytogenetic abnormalities in small cell lung cancer cell lines. AB - Ten cell lines were established from different biopsies from nine patients with small cell lung cancer (SCLC). These were established from metastases in the marrow (7), breast (1), pleural fluid (1), and spinal cord (1) and had been in culture for periods varying from 1 to 11 months. All cell lines exhibited typical cytological features of SCLC, and produced neuron specific enolase. All lines examined (5) contained dense core granules and were tumorigenic when injected intracranially into nude mice. None of the six cell lines tested had an amplified oncogene (c-myc or n-myc) previously reported to be amplified in SCLC. Detailed chromosome analyses were undertaken and showed that a structural abnormality of chromosome 3(p11p23) was a frequent (6 of 10) but not invariable finding. Our study and one other indicate that there is not a unique chromosome abnormality present in all cases of SCLC, although loss of chromosome 13 and structural abnormalities of chromosome 3 were frequently found. A feature of these SCLC cell lines established from metastatic deposits was the complexity of the karyotypes due to numerical and structural chromosomal abnormalities. PMID- 3034410 TI - Dissimilar effects of phorbol ester and diacylglycerol derivative on protein kinase activity in the monoblastoid U937 cell. AB - Mechanism, in addition to protein kinase C activation may mediate 12-O tetradecanoylphorbol-13-acetate (TPA) stimulated differentiation of leukemic cells. We compared the effect of pretreating intact monoblastoid U937 cells with TPA or the diacylglycerol derivative, 1-oleoyl-2-acetylglycerol (OAG), by studying the protein kinase C dependent and independent histone phosphotransferase activity, the phosphorylation of endogenous substrates, and the ability to stimulate differentiation. In cellular fractions derived from cells treated with TPA or OAG, cytosolic protein kinase C activity decreased. In the detergent extracted particulate fraction, TPA produced a time and dose dependent decrease in protein kinase C activity. In contrast, OAG increased particulate protein kinase C activity. In addition, the particulate fraction derived from cells treated with TPA exhibited increased phosphatidyl serine and diolein independent histone phosphotransferase activity as well as an increase in the phosphorylation of two endogenous substrates with molecular weights of 120,000 and 80,000. OAG did not mimic these effects. When exposed to 32P-labeled intact cells, OAG and TPA stimulated phosphorylation of three substrates. Thus, the inability of OAG to mimic the effects of TPA was not due to lack of protein kinase C activation. TPA, but not OAG, stimulated differentiation of the U937 cell to a monocyte-like cell. These data demonstrate that TPA and OAG have dissimilar effects on protein kinase activity and differentiation in the U937 monoblastoid cell. PMID- 3034411 TI - Levels of high energy phosphates in human lung cancer cell lines by 31P nuclear magnetic resonance spectroscopy. AB - Human lung cancers are divided into small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC) based on established criteria. SCLC differs from NSCLC by the expression of biomarkers, including creatine kinase-BB isoenzyme (EC 2.7.3.2). Subtypes of SCLC are referred to as classic and variant, both of which have elevated levels of creatine kinase-BB isoenzyme. We, therefore, applied 31P nuclear magnetic resonance spectroscopy to cell lines of classic SCLC, variant SCLC, and NSCLC human tumors, using continuous perfusion to identify any differences in the detectable levels of intracellular high-energy phosphate compounds. The spectra indicate that only the variant SCLC cells maintain high levels of phosphocreatine. Additionally, the classic SCLC cells express elevated levels of a diphosphodiester. Neither phosphocreatine nor diphosphodiesters are found in the NSCLC cell spectra. PMID- 3034412 TI - Formation of DNA interstrand cross-links by the novel chloroethylating agent 2 chloroethyl(methylsulfonyl)methanesulfonate: suppression by O6-alkylguanine-DNA alkyltransferase purified from human leukemic lymphoblasts. AB - The formation of DNA interstrand cross-links was compared in DNA treated with either 1,3-bis(2-chloroethyl)-1-nitrosourea or 2 chloroethyl(methylsulfonyl)methanesulfonate. DNA that was pulse treated briefly with either of these drugs continued to form cross-links at 37 degrees C for over 8 h after drug removal, indicating that such DNA contained stable precursors of cross-links. When human O6-alkylguanine-DNA alkyltransferase was added to the drug-treated DNA further cross-link formation was prevented at all points during this protracted time course, indicating that these stable cross-link precursors also remained substrates for this repair enzyme. Although the pattern of 2 chloroethyl(methylsulfonyl)methanesulfonate-induced cross-link formation and susceptibility to suppression by O6-alkylguanine-DNA alkyltransferase resembled that for 1,3-bis(2-chloroethyl)-1-nitrosourea, quantitative differences in the rates of cross-link formation and in the amounts of O6-alkylguanine-DNA alkyltransferase required to suppress cross-link formation suggest that critical differences exist between these agents. PMID- 3034413 TI - Induction of thymic lymphomas and squamous cell carcinomas following topic application of isopropyl methanesulfonate to female Hsd:(ICR)BR mice. AB - The goal of these experiments in female Hsd:(ICR)Br mice was to determine whether the direct-acting SN1 alkylating carcinogen isopropyl methanesulfonate (IMS) is carcinogenic and to compare its effects with those of the direct-acting SN2 methyl homologue, methyl methanesulfonate (MMS). The compounds were administered by topical application and s.c. injection. Analysis at the 288th day of mice receiving s.c. injections of IMS and MMS was the subject of a previous report (A. Segal et al., Proc. Soc. Exp. Biol. Med., 183: 132-135, 1986). The s.c. and topical application experiments were terminated at the 450th day and the final results are reported in this paper. In mice treated by s.c. injection with IMS, thymic lymphomas were observed in at least 20 of 32 mice, the first at the 40th day, and neoplasms were not observed at the injection site. Of the 30 MMS-treated mice, 11 developed sarcomas at the injection site and one thymic lymphoma was observed. In mice treated topically with IMS, thymic lymphomas were observed in 20 of 30 treated mice, the first at the 102nd day, and squamous cell carcinomas at the injection site were observed in 9 mice. Neither squamous cell carcinomas nor thymic lymphomas were observed in 30 mice following topical application of MMS. The direct-acting SN2 aklylating carcinogen beta-propiolactone was also administered by topical application. At the 450th day, at the same dose used for MMS (40 mumol/application), papillomas of the skin were observed in 25 of 30 treated mice, squamous cell carcinomas of the skin were seen in 17 mice, and one thymic lymphoma was observed. The results suggest that the rapid induction of thymomas by IMS may be related to its ability to alkylate exocyclic oxygen atoms in DNA of hemopoietic cells and also to a sensitivity of these cells to such lesions. PMID- 3034414 TI - Analysis of c-myc expression in a human hepatoma cell line. AB - A tumorigenic human hepatoma cell line, Hep G2, has been shown to have high steady-state levels of c-myc transcripts compared to normal human liver. We have now characterized c-myc expression in Hep G2 cells with regard to message stability, gene rearrangements, gene amplification, chromosomal translocations, promoter utilization, and the effects of protein synthesis inhibitors. We have determined that the half-life of the Hep G2 c-myc transcript is approximately 20 min and conclude that the high steady-state level of c-myc mRNA is not the result of a specific stabilization of the c-myc message but probably results from increased c-myc gene transcription. c-myc expression in Hep G2 cells appears to be constitutive, since it remains constant in different cell growth states (log phase versus nondividing cells). The high constitutive expression of the c-myc gene in Hep G2 cells could not be explained by gene amplification, gene rearrangements, or chromosomal translocations. However, based on an S1 nuclease protection assay, the P1/P2 promoter utilization ratio is approximately 1/1 which differs from the 1/5 P1/P2 ratio observed in normal human liver. Treatment with cycloheximide, a protein synthesis inhibitor, does not superinduce Hep G2 c-myc transcription based on transcription "run on" and RNA slot blot analysis. However, cycloheximide treatment does exert a posttranscriptional effect involving the specific stabilization of the c-myc message. PMID- 3034415 TI - Frequent expression of growth factors for mesenchymal cells in human mammary carcinoma cell lines. AB - The expression of growth factors for mesenchymal cells was investigated in 10 different human mammary carcinoma cell lines. Expression of mRNA for platelet derived growth factor (PDGF) A-chain, PDGF B-chain, transforming growth factor alpha, and insulin-like growth factor II were found in eight, nine, five, and two of the cell lines, respectively. The production of PDGF-like growth factors by the mammary carcinoma cell lines was investigated in detail. PDGF receptor competing activity and mitogenic activity, which could be neutralized by PDGF antibodies, were found in the conditioned medium of almost all the cell lines. A PDGF-like growth factor was partially purified from the conditioned medium of one of the cell lines, MCF7. A Mr 31,000 component, which was reduced to Mr 17,000, was furthermore precipitated by a PDGF antiserum from the conditioned medium of metabolically labeled MCF7 cells. These results indicate that growth factors for mesenchymal cells are frequently expressed in human mammary carcinoma cell lines. This is of interest in relation to the fact that mammary carcinoma tumors often contain a high proportion of proliferating connective tissue cells. PMID- 3034416 TI - Aflatoxin exposure measured by urinary excretion of aflatoxin B1-guanine adduct and hepatitis B virus infection in areas with different liver cancer incidence in Kenya. AB - Two major etiological agents, hepatitis B virus and aflatoxin B1, are considered to be involved in the induction of liver cancer in Africa. In order to elucidate any synergistic effect of these two agents we conducted a study in various parts of Kenya with different liver cancer incidence in order to establish the rate of exposure to aflatoxin and the prevalence of hepatitis infections. Of all tested individuals 12.6% were positive for aflatoxin exposure as indicated by the urinary excretion of aflatoxin B1-guanine. Assuming no annual and seasonal variation, a regional variation in the exposure was observed. The highest rate of aflatoxin exposure was found in the Western Highlands and Central Province. The incidence of hepatitis infection nationwide as measured by the presence of the surface antigens was 10.6%, but a wide regional variation was observed. A multiplicative and additive regression analysis to investigate if hepatitis and aflatoxin exposure had a synergetic effect in the induction of liver cancer was negative. However, a moderate degree of correlation between the exposure to aflatoxin and liver cancer was observed when the study was limited to certain ethnic groups. The study gives additional support to the hypothesis that aflatoxin is a human liver carcinogen. PMID- 3034417 TI - Antioxidant enzymes and survival of normal and simian virus 40-transformed mouse embryo cells after hyperthermia. AB - Relative cell survival and activity of the free radical scavenging enzymes superoxide dismutase, catalase, and glutathione peroxidase were measured in cloned normal (MEA) and SV40-transformed (SVMEA) mouse embryo cells exposed at 44 degrees C for 0-3 h. At 37 degrees C, all three enzymes were 2-5 times higher in MEA than in SVMEA. Hyperthermia did not significantly alter enzyme levels in either cell line but selectively reduced transformed cell survival to less than 5% while relative survival of normal cells remained above 75%. The latter, however, could be reduced to 25% when normal cells were pretreated with 3 mM diethyldithiocarbamate, an inhibitor of copper- and zinc-containing superoxide dismutase. Similar treatment rendered SVMEA extremely thermosensitive. On the other hand, sublethal heat treatment (15 min at 45 degrees C) of cultured cells resulted in a relative thermal resistance upon subsequent exposure to 45 degrees C for 1-4 h. This induced thermotolerance was associated with a rise in antioxidant enzyme levels and both became significant only 4-6 h after the initial heat treatment. Induced enzyme and thermotolerance levels in transformed cells remained, nonetheless, far below those of normal cells. The data show that inherent (in MEA) as well as induced (in SVMEA) thermotolerance is associated with high antioxidant enzyme levels while the reverse is true in the case of inherent (in SVMEA) and induced (in MEA) thermosensitivity. These findings suggest that increased production of oxygen free radicals may be involved in hyperthermic cell injury, which then becomes a function of basal or inducible levels of antioxidant enzymes. Induction of the latter by hyperthermia is apparently inefficient in transformed cells making them more vulnerable. Enzyme induction seems also to require a lag period of 4-6 h suggesting the possible involvement of an intermediate inducer(s) at molecular level. The so-called heat shock proteins may be candidates for such a role. PMID- 3034418 TI - Enhanced activity of mouse peritoneal cells after aclacinomycin administration. AB - We have investigated the activation of mouse peritoneal macrophages by injection of aclacinomycin (ACM). Macrophages from ACM-treated mice have an increased phagocytic activity as measured by Candida ingestion. The microbicidal activity indirectly evaluated by chemiluminescence and superoxide determination in response to stimulation with zymosan and 4 beta-phorbol-12-myristate-13 alpha acetate is also greater in the cells from treated mice. Direct measurement of the cytostatic function, and of in vitro and in vivo cytotoxicity shows comparable significant increases against L1210 or P815 target cells. The enhanced antitumoral activity could not be attributed to the residual presence of ACM in the peritoneal cells since no drug was detected by high-performance liquid chromatography and since their freeze-thaw lysates incubated with P815 cells did not modify the growth of tumor cells as measured by [3H]-thymidine incorporation. We also checked that the presence of ACM did not influence the intensity of the chemiluminescence. In all tests performed, only i.p. ACM administration could stimulate the peritoneal cells. Since the doses of ACM inducing an increase in macrophage activity are effective on the survival of tumor-bearing mice, the participation of this mechanism in tumor control might be suggested. PMID- 3034419 TI - Early spontaneous deficiency of calcitonin renal binding sites in rats with a high incidence of calcitonin-secreting tumors (WAG/Rij). AB - Old rats of the WAG/Rij strain have a high incidence (50%) of medullary thyroid carcinoma, a calcitonin (CT)-secreting tumor. We have characterized and quantified the topographical distribution of [125I]salmon calcitonin (sCT) binding sites in the kidneys of this strain, as compared to Wistar CF rats (2% incidence of spontaneous medullary thyroid carcinoma). We report here that, up to 15 days of postnatal development, the distribution of CT-binding sites in the kidney of the WAG/Rij strain was quite similar to that found in developing and adult Wistar CF rats. However, from the age of 1 month, sCT-binding sites were dramatically reduced in both the medulla and the inner part of the kidney cortex, though plasma CT levels were not significantly different in both strains. Adult WAG/Rij rats bearing a transplanted tumor for 12 weeks had a high level of plasma calcitonin and exhibited an even greater reduction of both medullary and cortical sCT-binding sites. These results suggest that the modification in the CT-binding sites in WAG/Rij rats is not a consequence of a possible down regulation due to elevated circulating hormonal level but could be inherited and possibly associated with the later development of the tumor in this strain. PMID- 3034420 TI - Oestradiol 16 alpha-hydroxylase: a risk marker for breast cancer. AB - 17-oxidation and 2- and 16 alpha-hydroxylation of oestradiol show important correlations with biological aspects of breast cancer in women and mice. The presence of mouse mammary tumour virus may lead to increased 16 alpha hydroxylation. 16 alpha-hydroxyoestrone (which may be generated in an autocrine manner) forms covalent linkages with amino groups on proteins and nucleotides. PMID- 3034421 TI - Dietary factors and breast cancer. AB - Population and case-control studies indicate the involvement of diet in breast cancer aetiology. Obesity appears to be a risk factor for postmenopausal disease, and most likely operates through the modification of oestrogen synthesis and metabolism. Dietary fat, though a major source of calories and, therefore, a contributor to the development of obesity, may have a more specific role, involving oestrogens, protein hormones and growth factors, or other, non endocrine, mechanisms. Obesity also has an adverse effect on breast cancer prognosis after surgery. Again, this may, at least partially, involve enhanced oestrogenic stimulation of residual tumours in postmenopausal patients. Though knowledge of these relationships can be expanded by the application of appropriate animal models, the final test has to be the implementation of carefully designed intervention trials in groups of women at high risk of breast cancer, and as an adjunct to surgery in breast cancer patients. PMID- 3034422 TI - Phase II trial evaluating continuous infusion of etoposide, cisplatin, and hexamethylmelamine in extensive-disease small cell carcinoma of the lung. AB - A total of 27 untreated and 24 previously treated patients with extensive-disease small cell lung cancer (SCLC) were treated with a combination chemotherapeutic regimen of continuous-infusion etoposide for 5 days, cisplatin, and hexamethylmelamine. of 25 evaluable patients with untreated SCLC, three (12%) achieved a complete response and 16 (64%) achieved a partial response. Among 23 evaluable patients with relapsed SCLC there were no complete responses and nine (39%) achieved a partial response. Median survival times were 252 and 109 days for the above groups, respectively. Myelotoxicity, especially thrombocytopenia, was moderately severe. Other toxic effects, including renal and neurologic, were minimal. These results compare favorably with other regimens including etoposide and cisplatin. Our results further confirm the activity of etoposide and cisplatin as both initial therapy and as a salvage regimen in the management of patients with extensive-stage SCLC. PMID- 3034423 TI - Phase II trial of etoposide in the management of advanced or recurrent mixed mesodermal sarcomas of the uterus: a Gynecologic Oncology Group Study. PMID- 3034424 TI - High-dose (200 mg/m2) cisplatin-induced neurotoxicity in primary advanced ovarian cancer patients. PMID- 3034425 TI - Applications of hydroxylapatite in oral and maxillofacial surgery. Part II: Ridge augmentation and repair of major oral defects. PMID- 3034426 TI - Hydrogen peroxide and salt solutions: are they effective antiplaque agents? PMID- 3034427 TI - [The role of Epstein-Barr viruses in the etiopathogenesis of rheumatoid arthritis]. PMID- 3034428 TI - [Cleft lip and palate: relation to cytomegalovirus infection]. PMID- 3034430 TI - The proton flux through the bacterial flagellar motor. AB - Bacterial flagella are driven by a rotary motor that utilizes the free energy stored in the electrochemical proton gradient across the cytoplasmic membrane to do mechanical work. The flux of protons coupled to motor rotation was measured in Streptococcus and found to be directly proportional to motor speed. This supports the hypothesis that the movement of protons through the motor is tightly coupled to the rotation of its flagellar filament. Under this assumption the efficiency of energy conversion is close to unity at the low speeds encountered in tethered cells but only a few percent at the high speeds encountered in swimming cells. This difference appears to be due to dissipation by processes internal to the motor. The efficiency at high speeds exhibits a steep temperature dependence and a sizable deuterium solvent isotope effect. PMID- 3034429 TI - lac repressor can regulate expression from a hybrid SV40 early promoter containing a lac operator in animal cells. AB - The E. coli lac operator and repressor were adapted for function in mammalian cells. Plasmids containing an SV40 early region (pSVlacO) or a chloramphenicol acetyl transferase gene (pSVlacOCAT) linked to a hybrid SV40 early promoter bearing a lac operator were tested for function. Identical plasmids lacking an operator (pX-8 and pX-8CAT) were controls. In vitro, early transcription from pSVlacO, but not from pX-8, was inhibited by lac repressor, and repression was overcome by IPTG. Repression of large T synthesis or CAT activity occurred in vivo only when the respective operator-containing plasmid was cotransfected with a plasmid encoding lac repressor, or when the recipient cells stably synthesized lac repressor. IPTG substantially relieved repression in both cases. CAT enzyme repression was paralleled by a decrease in CAT mRNA abundance. Thus regulatory elements of the lac operon function physiologically in mammalian cells. PMID- 3034431 TI - Phosphodiester bond cleavage outside mitochondria is required for the completion of protein import into the mitochondrial matrix. AB - The present studies show that hydrolysis of a phosphodiester bond, most likely ATP, is a distinct, second step required to complete import of the F1-ATPase beta subunit into the mitochondria. This step follows a membrane potential-dependent first step. We show, using an inhibitor of adenine nucleotide transport and the analogue beta,gamma-AMP-PCP, that the activity required for this phosphodiester hydrolysis-dependent completion of protein import resides outside the mitochondrial inner membrane. This activity is proposed to act on the precursor at the site of translocation either to render it competent or to catalyze its vectorial movement directly through the import apparatus. This activity shares properties ascribed to proteins of the heat-shock family, which are proposed to participate in the ATP-dependent refolding of partially denatured proteins and nascent peptides. PMID- 3034432 TI - Phorbol ester-inducible genes contain a common cis element recognized by a TPA modulated trans-acting factor. AB - The promoter regions of several phorbol diester-(TPA-) inducible genes (collagenase, stromelysin, hMT IIA, and SV40) share a conserved 9 bp motif. Synthetic copies of these closely related sequences conferred TPA inducibility upon heterologous promoters. Footprinting analysis indicated that these TPA responsive elements (TREs) are recognized by a common cellular protein: the previously described transcription factor AP-1. A point mutation that eliminated the basal and induced activity of the TRE also interfered with its ability to bind AP-1. Treatment of cultured cells with TPA led to a rapid 3- to 4-fold increase in TRE binding activity, by a posttranslational mechanism. These results strongly suggest that AP-1 is at the receiving end of a complex pathway responsible for transmitting the effects of phorbol ester tumor promoters from the plasma membrane to the transcriptional machinery. PMID- 3034433 TI - Purified transcription factor AP-1 interacts with TPA-inducible enhancer elements. AB - The enhancer-binding protein AP-1 has been purified to greater than 95% homogeneity from HeLa cells by sequence-specific DNA affinity chromatography and identified as a 47 kd polypeptide. Purified AP-1 activates transcription in vitro of the wild-type human metallothionein IIA (hMT IIA) gene but not mutant hMT IIA promoters lacking AP-1 recognition sites. DNAase I protection analysis indicates that genetically defined enhancer elements in hMT IIA, SV40, and the human collagenase gene contain high-affinity AP-1-binding sites, each with a conserved recognition motif, TGACTCA. These three genes are transcriptionally induced by treatment of cells with the tumor promoter TPA. Here we demonstrate that multiple synthetic copies of the consensus AP-1-binding site can act as TPA-inducible enhancers in various plasmid constructs after transfection into HeLa cells. These findings suggest that AP-1 is a transcription factor that functions by interacting with a specific enhancer element, and that its activities may be modulated by treatment of cells with TPA, known to stimulate protein kinase C. PMID- 3034434 TI - The role of the L3T4 molecule in mitogen and antigen-activated signal transduction. AB - We investigated the role of the L3T4 molecule in mitogen and antigen-initiated signal transduction in the L3T4(+) murine T cell hybridoma, 3DT52.5.9 and an L3T4(-) variant, 3DT52.5.24. Both Concanavalin A (Con A) and specific antigen stimulated increases in cytosolic-free calcium ([Ca2+]i), phosphatidylinositol turnover, and interleukin-2 (IL-2) production in both cell lines. About 85% of the stimulated rise in [Ca2+]i was from an extracellular source. Anti-L3T4 monoclonal antibody (MAb) inhibited 90% of antigen- and 50% of Con A-stimulated increases in [Ca2+]i and IL-2 production but had no effect on the ability of either activation signal to stimulate phosphatidylinositol turnover in the parent L3T4(+) cells. Stimulus-response coupling in the L3T4(-) cells was unaffected by the MAb. The anti-L3T4-insensitive increase in [Ca2+]i induced by Con A was inhibited by EGTA, suggesting that this mitogen also stimulated an influx of Ca2+ via an additional transport mechanism distinct from that stimulated by antigen. The fact that anti-L3T4 antibodies inhibit antigen and Con A-stimulated Ca2+ transport and IL-2 production without affecting phosphatidylinositol turnover suggests that L3T4 may play a critical role in modulating the activation of the T cell receptor-associated Ca2+ transporter in T cell stimulus-response coupling. PMID- 3034435 TI - The carboxyl terminus of the hamster beta-adrenergic receptor expressed in mouse L cells is not required for receptor sequestration. AB - The structural basis for agonist-mediated sequestration and desensitization of the beta-adrenergic receptor (beta AR) was examined by oligonucleotide-directed mutagenesis of the hamster beta AR gene and expression of the mutant genes in mouse L cells. Treatment of these cells with the agonist isoproterenol corresponded to a desensitization of beta AR activity. A mutant receptor that bound agonist but did not couple to adenylate cyclase showed a dramatically reduced sequestration response to agonist stimulation. In contrast, beta AR mutants in which the C-terminus was truncated and/or in which two regions that have been proposed as phosphorylation substrates for cAMP-dependent protein kinase were removed showed normal sequestration responses. These results demonstrate that agonist-mediated sequestration of the beta AR can occur in the absence of the C-terminus of the protein and reveal a strong correlation between effective coupling to Gs and sequestration. PMID- 3034436 TI - [Cystosarcoma phyllodes]. PMID- 3034437 TI - [Pulmonary adenomatosis]. PMID- 3034438 TI - Effect of phenobarbital and 3-methylcholanthrene on aldehyde dehydrogenase activity in cultures of HepG2 cells and normal human hepatocytes. AB - Aldehyde dehydrogenase (ALDH) activity was measured in primary cultures of normal human hepatocytes and of the human hepatoma cell line HepG2 after application of phenobarbital (PB) or 3-methylcholanthrene (MC) for 5 days. Treatment with PB alone resulted in a significant increase in both protein and DNA content at concentrations of 2 and 3 mM. Treatment with MC at a concentration as low as 5 microM led to a significant loss of cells when it lasted more than 5 days. Concentrations of 3-5 mM of PB in the media of HepG2 cell cultures caused a 2 fold enhancement of the activity of ALDH, as measured with NAD and propionaldehyde (P/NAD) or benzaldehyde (B/NAD). On the other hand, MC-treated cultures (5 microM) showed a 20-fold increase in enzyme activity measured with NADP and benzaldehyde (B/NADP), and a 2-fold increase in B/NAD activity. Combined treatment with both PB and MC led to an effect of dynamic synergism as far as B/NAD and B/NADP activities are concerned, suggesting a metabolite of MC as the mediator for the increase of ALDH activity. Normal human hepatocytes in primary cultures responded to PB (3 mM) in a similar way as HepG2 cells as far as DNA and protein content and ALDH activity are concerned. It is concluded, that HepG2 hepatoma cells behave similar to the normal hepatocytes in terms of ALDH regulation and can be used for studies on the activity of ALDH as modified by added xenobiotics. PMID- 3034439 TI - Hydroxyl radical production in the presence of fibres by a Fenton-type reaction. AB - Glass fibres are considered to be inert and therefore thought to present no real hazard to the health of people who inhale them. Results in the present study however indicate that these fibres are able to produce hydroxyl radicals in the presence of hydrogen peroxide by a Fenton-type reaction. Since hydroxyl radical is implicated in lipid peroxidation, single-strand DNA breaks and carcinogenesis, care should be exercised when dealing with glass fibres. PMID- 3034440 TI - [Study on histogenesis, histological types and prognosis of 100 cases of gastric carcinoma]. PMID- 3034441 TI - [Trial treatment of viral keratoconjunctivitis]. PMID- 3034442 TI - Immune status of the population to poliovirus infection in Singapore. AB - A seroepidemiological survey conducted after two decades of mass immunization against poliomyelitis in Singapore showed a high level of herd immunity in the population to all three types of human poliovirus. Neutralizing antibodies to poliovirus 1, 2 and 3 were detected in 93.9-96.7% of the population between 6 months and over 40 years old. Adults above 30 years of age had a relatively high level of susceptibility to poliovirus infection, with 10.2-12% in the over-30 year age group possessing no neutralizing antibody against poliovirus 1, 14.3% in the 40+ age group against poliovirus 2, and 14% in the 30-39-year age group against poliovirus 3. The geometric mean titres showed a general decline among the older age groups. There were no statistically significant differences in the immune status by sex and by ethnic groups (Chinese, Malays and Indians). The importance of maintaining a high coverage of immunization, monitoring the herd immunity of the population to identify high-risk groups, and advising adults travelling to the endemic areas to be immunized against poliomyelitis was stressed. PMID- 3034443 TI - Intra-arterial administration of epirubicin in the treatment of nonresectable hepatocellular carcinoma. Epirubicin Study Group for Hepatocellular Carcinoma. AB - A group study was conducted to investigate the effect of intrahepatic arterial administration of epirubicin in the treatment of nonresectable hepatocellular carcinoma (HCC). Sixty-four patients entered the study. There were 51 men and 13 women. The age range was from 32 to 79 years, with an average of 59.1. Fifty-four patients had associated cirrhosis of the liver. Epirubicin in a dose of 60-90 mg/m2 was infused as a bolus into the hepatic artery 1-4 times (average 1.8) at intervals of 3 weeks to 3 months. Tumor size was properly evaluated in 53 patients. There were 1 CR (complete responses), 7 PR (partial responses), 34 NC (no change), and 11 PD (progression of disease). Thus, the response rate (CR + PR) was 15.1%. Seventeen patients are still alive 305-730 days (mean 505 days) after the initial treatment. A higher dose and more treatment courses tended to produce a better result. The most common side effects of this drug were bone marrow suppression, gastrointestinal symptoms, and alopecia. Cardiac toxicity was not observed with the doses used in this study. A retrospective comparison of the present result with that of patients treated by intra-arterial administration of doxorubicin demonstrated that epirubicin is more effective than doxorubicin in teams of survival rate. PMID- 3034445 TI - 'Water structure' versus 'radical scavenger' theories as explanations for the suppressive effects of DMSO and related compounds on radiation-induced transformation in vitro. AB - We report here that dimethylsulfoxide (DMSO): suppresses radiation-induced transformation in vitro, even when DMSO treatments begin as late as 10 days post irradiation (when cells are in the confluent, stationary phase of growth); inhibits the 12-O-tetradecanoylphorbol-13-acetate (TPA) enhancement of radiation induced transformation in vitro; does not affect the expression of transformed cells as foci (when surrounded by non-transformed cells); and may be affecting radiation-induced transformation through its solvent properties (i.e. the 'Water Structure' theory), while its effects on the TPA enhancement of radiation transformation may be mediated by its free radical scavenging abilities. DMSO, dimethylformamide (DMF) and dimethylacetamide (DMA) are similar solvents which are all very effective in their ability to suppress radiation-induced transformation in vitro (at concentrations in the cellular media down to 0.01%). As DMSO is known to be an extremely effective OH. free-radical scavenging agent, while DMF and DMA are not as efficient at scavenging free radicals, our results suggest that properties other than free-radical scavenging ability may be important in the suppressive effects of these compounds on radiation-induced transformation in vitro. It is known that low concentrations of such basic aprotic solvents modify water structure so as to suppress the protic (H-bond donor) reactivity of water and enhance its basic (H-bond receptor) reactivity. These reactivity changes may well be responsible for the effects noted above. DMSO, DMF and DMA are also capable of suppressing the TPA enhancement of radiation transformation (at concentrations of the compounds of 0.1% or higher). For this effect, the ability of these compounds to scavenge OH. shows a general correlation with their ability to suppress the TPA enhancement of transformation, suggesting that the 'Radical Scavenger' theory may explain the ability of DMSO to suppress promotion in vitro. PMID- 3034444 TI - In vitro colony inhibition of carboplatin against stomach and lung cancer cell lines in comparison with cisplatin. AB - The effects of carboplatin and cisplatin on colony formation in stomach and lung cancer cell lines were examined and compared. The colony-inhibitory activity of carboplatin against stomach and lung cancer cell lines was similar to that of cisplatin when one-tenth of the peak plasma concentration of each drug was used (r = 0.80). One of the four stomach cancer cell lines was sensitive to carboplatin although all the stomach cancer cell lines were resistant to cisplatin. Of the three small cell lung cancer cell lines tested, two were sensitive to both carboplatin and cisplatin, and only one cell line (N857) was resistant to cisplatin; all the non-small cell cancer cell lines tested were resistant to both drugs. On the basis of these preliminary results, we suggest that carboplatin has potential therapeutic activity against stomach cancer and should be evaluated carefully from this aspect. PMID- 3034446 TI - Subcellular calmodulin distribution in rat liver after CCl4 poisoning. AB - Disturbed cellular calcium homeostasis has been observed during CCl4 poisoning, with an increase in calcium content 1 h after administration. Intracellular increase of calcium may be expected to alter membrane/cytosol distribution of calmodulin (CaM). This paper investigates changes in rat liver subcellular CaM distribution 30 min, 1 h and 2 h after CCl4 intoxication. The whole liver value remained unchanged, whereas the nuclear fraction increased and the microsomal and cytosolic fraction decreased. This may suggest that CaM is involved in the several liver cell alterations caused by CCl4 poisoning. PMID- 3034447 TI - Biochemical and immunochemical determination of (Na+ + K+)-ATPase content in subcellular fractions prepared from the avian salt gland: evidence for enzyme heterogeneity. AB - Subcellular membrane fractions were prepared from the salt glands of osmotically stressed ducklings. Two fractions were characterized biochemically with respect to (Na+ + K+)-ATPase, alkaline phosphodiesterase I, succinate dehydrogenase, esterase, and galactosyltransferase activities and immunochemically with respect to (Na+ + K+)-ATPase. The ratios of the estimates of the (Na+ + K+)-ATPase contents obtained biochemically and immunochemically from the two fractions differed by more than 2 X. The results are consistent with the presence of at least two molecular species of (Na+ + K+)-ATPase, unevenly distributed between the two fractions. PMID- 3034448 TI - Effects of procaine on pharmaco-mechanical coupling mechanisms activated by acetylcholine in smooth muscle cells of porcine coronary artery. AB - The action of procaine on pharmaco-mechanical coupling activated by application of acetylcholine (ACh) was investigated using collagenase-treated dispersed intact and skinned smooth muscle cells and intact muscle tissues of the porcine coronary artery. ACh reduced stored 45Ca2+, and this action was prevented by procaine in intact dispersed cells. The maximum reduction in the level of stored 45Ca induced by caffeine (25 mM) or inositol 1,4,5-trisphosphate (InsP3; 3 microM) was also prevented by procaine in the skinned muscle cells in the presence or absence of ATP. However, inhibitions of the latter required higher concentrations of procaine than the former. Release by 10 microM ACh of Ca2+ from its store site in the presence or absence of extracellular Ca2+ was also inhibited by procaine and was detected using the quin2 fluorescence method. In these smooth muscle tissues, ACh (above 10 nM) reduced the amount of phosphatidylinositol 4,5-bisphosphate (PI-P2) and dose dependently increased the amount of phosphatidic acid. Procaine inhibited the hydrolysis of PI-P2 activated by ACh, thus reducing the amount of InsP3 and the release of Ca2+ from the store site. It is concluded that procaine has multiple actions on the porcine coronary artery, and one of the actions related with pharmacomechanical coupling appears through inhibition of hydrolysis of PI-P2 induced by ACh. PMID- 3034449 TI - Isozyme specific modification of myosin ATPase by cAMP in rat heart. AB - The total ATPase activity of myosin and the values for the isozyme V1 have been measured in hearts from rats of different ages and with different levels of thyroid function. The contribution of V3 was calculated from the difference between total and V1 ATPase, neglecting the small contribution of V2. Hearts were quickly frozen after rapid removal from the animals in order to preserve the state of ATPase activity that existed in the intact animal, and ATPase activity was measured in thin sections of tissue by a microphotometric technique. In euthyroid hearts, although cAMP increases total myosin ATPase activity and the activity of V1, the cyclic nucleotide inhibits the ATPase activity of V3. In hearts from rats with developing hypothyroidism following thyroidectomy, the same occurs. After a sufficient period has elapsed after thyroidectomy for V1 to have practically disappeared, cAMP has no effect on ATPase activity, but the injection of thyroid hormone restores the effect. Total myosin ATPase activity is maintained relatively constant as the animal ages from 80 to 165 days and during the first 10-11 days following thyroidectomy even though the concentration of V1 is dropping. The explanation proposed for these observations is that myosin can exist in two different forms, only one of which can participate in the active generation of force. The transition between the two forms is regulated by a soluble factor that is itself controlled by the adrenergic system. The factor(s) involved in this regulatory mechanism is soluble and can be transferred between different thin sections cut from a frozen heart. PMID- 3034450 TI - Effect of nedocromil sodium on adenosine- and methacholine-induced bronchospasm in asthma. AB - The effect of nedocromil sodium on adenosine-induced bronchospasm was investigated in eight asthmatic patients. Nedocromil sodium (4 mg) administered by aerosol 10 min before challenge, effectively inhibited adenosine-induced bronchospasm. In another group of six asthmatic patients, nedocromil sodium administered in the same way did not reduce the bronchial response to methacholine. These results support the view that adenosine-induced bronchospasm may be prevented by a membrane stabilizing drug and is not mediated through cholinergic pathways. The mechanism(s) by which nedocromil sodium inhibits adenosine-induced bronchospasm requires further investigations. PMID- 3034451 TI - Phosphatidylethanolamine methyltransferase and cAMP, cGMP phosphodiesterases in lymphocytes and monocytes in sarcoidosis. AB - Among the various hypotheses proposed to explain immune cell defect in sarcoidosis, we examined thoroughly that of Faguet who described abnormalities of signal transmission at lymphocyte membrane level. Phosphatidylethanolamine methyltransferase and cAMP cGMP phosphodiesterases were studied in blood lymphocytes and monocytes from 8 subjects with sarcoidosis disease. Phosphatidylethanolamine methyltransferase (PMT1) plays an important regulatory role in membrane signal transmission. cAMP and cGMP phosphodiesterases (PDE) regulate cytoplasmic cyclic nucleotide levels and so participate in the modulation of the cell cycle. We observed a decreased PMT1 activity in lymphocytes and monocytes and a decreased cAMP and cGMP PDE activities in monocytes. It is not now possible to say if these abnormalities are primary or secondary. Whatever the origin of this dysfunctioning, these results evoke simultaneous disturbances of membrane signal transmission and cell cycle in monocytes and membrane abnormalities in lymphocytes. These abnormalities could explain some immune cell defects in sarcoidosis disease. PMID- 3034452 TI - Measurement by bioluminescence technique of erythrocyte membrane Na+,K+-ATPase activity in hypertensive patients. AB - Erythrocyte membrane Na+,K+-ATPase activity was measured using a bioluminescence technique in 28 hypertensive patients (24 with essential hypertension, 2 with renovascular hypertension and 2 with hypertension secondary to primary hyperaldosteronism) and in 28 normotensive control subjects matched for age and sex. Erythrocyte Na+,K+-ATPase activity was significantly reduced in the patients with essential hypertension (130.9 +/- 11.4 vs. 186.6 +/- 19.5 nmol ATP/mg prot per h; mean values +/- SEM; p less than 0.05) and in the patients with secondary hypertension. A significant negative correlation was found between erythrocyte Na+,K+-ATPase and systolic blood pressure (r = -0.603; p less than 0.01), but not between Na+,K+-ATPase and plasma renin activity or plasma aldosterone levels. These data confirm the findings of a number of previous studies reporting reduced activity of erythrocyte Na+,K+-ATPase possibly related to the presence of a circulatory inhibitor of sodium pump. The method, based on ATP assay by bioluminescence, presents a high degree of specificity as well as simple, rapid execution. PMID- 3034453 TI - Detection of desialylated forms of human chorionic gonadotropin. AB - Urinary forms of human chorionic gonadotropin (hCG) with oligosaccharides deficient in sialic acid content (ashCG) have been reported to be excreted by patients with choriocarcinoma in greater amounts than by healthy, pregnant women. Although ashCG potentially could be a useful marker for the diagnosis and management of gestational trophoblastic neoplasia, the methods previously used for its detection were not suitable for routine clinical application. Therefore, we have devised a simpler method which can provide specific and sensitive measurements of ashCG in urine. This method, which is designated as a lectin immunoradiometric assay (LIRMA), employs an agarose-coupled lectin to selectively extract the ashCG, which is then quantified directly with a purified and radiolabelled rabbit antibody. The LIRMA has been applied to demonstrate that there is an increased excretion of ashCG by choriocarcinoma patients. It is also applicable, in principle, for the study of any glycoprotein which has a reduced content of sialic acid in its carbohydrate side chains. PMID- 3034454 TI - Pyrimidine 5'-nucleotidase in human granulocytes and platelets. PMID- 3034456 TI - Atrial natriuretic peptide and the natriuresis due to acute beta-blockade in conscious hypertensive rats. AB - Acute beta-adrenoceptor blockade in rats is known to produce a natriuresis of hitherto uncertain cause. To investigate this phenomenon plasma atrial natriuretic peptide (ANP) concentrations were measured in groups of conscious metoprolol-treated and control rats. In the active treatment group the blood pressure decreased slowly, as expected, settling at a lower level after 2 hours, and the mean sodium excretion doubled 1 hour after metoprolol administration. This natriuretic effect was maximal after 40-60 minutes and thereafter slowly declined towards basal values. The period of enhanced sodium excretion was associated with a significant rise (68%) of the mean ANP plasma concentration. It suggested that this increase in plasma ANP concentration can mediate the acute natriuretic effect beta-adrenoceptor blockade. PMID- 3034455 TI - Determination of creatine kinase isozymes in sera and tissues of patients with various lung carcinomas. AB - Three creatine kinase isozymes (CK-BB, CK-MB and CK-MM) were estimated by immunoassay in tumor tissues and in sera of patients with various lung carcinomas. CK-BB was increased in small cell carcinoma, but not in other lung carcinomas. CK-MM and CK-MB were not increased in any types of carcinoma. Serum CK-BB was increased in all types of lung carcinoma examined, while serum CK-MM and CK-MB were within normal limits in all patients. Serum CK-BB of healthy adults was estimated as 0.32 +/- 0.14 (mean +/- SD) ng/ml, ranging from 0.11-0.68 ng/ml. If CK-BB values above 1.0 ng/ml were considered abnormal, elevation occurred in 28/40 (70%) of patients with small cell carcinoma, 25/67 (37%) with adenocarcinoma, 21/51 (41%) with squamous cell carcinoma, 4/11 (36%) with other carcinoma of the lung and 10/42 (24%) with lung tuberculosis. Since serum CK-BB with lung cancer changed in parallel with the clinical course, this isozyme may be a marker for monitoring the clinical course, especially in small cell carcinoma of the lung. PMID- 3034458 TI - Mohs' surgery. New concepts and applications. AB - During the past decade, Mohs' surgery has been used increasingly, mostly by dermatologists, for the treatment of cutaneous neoplasms, usually basal cell or squamous cell carcinomas. This use has given rise to new developments, both in the technique itself and in basic tumor biology. As more tumors are delineated microscopically in both their histologic and growth patterns, certain concepts are emerging that are important for successful management of skin cancer. PMID- 3034457 TI - Expression of a germline human kappa chain-associated cross-reactive idiotype after in vitro and in vivo infection with Epstein-Barr virus. AB - The mouse monoclonal antibody 17.109 recognizes a cross-reactive idiotype (CRI) associated with kappa IIIb light chains of human IgM-rheumatoid factor (RF) paraproteins. The 17.109 idiotypic determinant is encoded by one or a group of closely related V kappa genes. The association of the idiotype with IgM- and IgA rheumatoid factors in certain autoimmune diseases necessitates an understanding of how human B lymphocytes can be induced to express the idiotype. To investigate the cellular expression of the 17.109 CRI, peripheral blood lymphocytes from normal donors were stimulated in vitro with Epstein-Barr virus (EBV) and pokeweed mitogen (PWM). EBV induced greater expression of IgM-associated 17.109 CRI than did PWM. The 17.109 CRI was preferentially associated with IgM rather than with IgG. In vivo EBV infection was studied in college students with infectious mononucleosis and displayed similar elevation of IgM-associated 17.109 CRI in sera obtained at presentation of clinical illness. Later, IgM levels declined while IgG-associated 17.109 CRI rose. The 17.109 idiotype was unrelated to antibodies against the Epstein-Barr virus nuclear antigen and the viral capsid antigen and was probably due to generalized activation of early B cells. These observations support the hypothesis that the 17.109 CRI is expressed by in vitro and in vivo EBV-infected cells. The 17.109 idiotype identifies a highly conserved V kappa gene product, which is expressed preferentially after EBV infection, but not exclusively with RF autoantibodies. PMID- 3034459 TI - Plasma endogenous digitalis-like factors in healthy individuals and in dialysis dependent and kidney transplant patients. AB - Plasma digitalis-like factors (DLF), dehydroepiandrosterone sulfate (DHEAS) and cortisol were assayed in 20 healthy subjects, 22 dialysis dependent subjects and 30 patients with kidney transplants. DLF were assayed on plasma extracts by digoxin radioimmunoassay (RIA), and by Na,K-ATPase inhibition and [3H]-ouabain displacement using hogbrain Na,K-ATPase. Values for the 3 methods strongly intercorrelated (r = 0.99, p less than .001). Mean values for plasma DLF, assayed by all three methods, were significantly greater in dialysis dependent subjects, than in healthy subjects (p less than .0001). Mean plasma DLF values measured by digoxin RIA in renal transplant recipients, were significantly lower than in dialysis dependent subjects (p less than 0.0001) and higher than in healthy subjects. Plasma DLF values correlated inversely with creatinine clearance (p less than 0.01). Plasma DHEAS levels were significantly lower and contributed substantially less to digoxin antibody reactivity and ouabain displacement in dialysis subjects and in renal transplants compared with healthy subjects. There was no change in plasma immunoreactive DLF, DHEAS or cortisol measured before and after dialysis. DHEAS is a major digoxin like immunoreactive DLF and a minor Na,K ATPase inhibitor in healthy subjects but makes only a minor contribution to DLF in dialysis and transplant subjects. We found the assays involving Na,K-ATPase inhibition and [3H]-ouabain displacement from Na,K-ATPase to be more sensitive for plasma DLF than the digoxin RIA, but because of the strong correlation between the methods, we suggest the RIA on plasma extracts can be used as a screening procedure. PMID- 3034460 TI - Cellular and molecular bases for dystocia. AB - In this review I have briefly outlined some of the cellular and molecular reasons for dystocia. I have described the myogenic, that may control the normal progression of labor and explain inadequate uterine contractility associated with dystocia. I have placed particular emphasis on our studies of gap junctions and their regulation. These studies show that there are specific physiologic mechanisms for regulating structural and functional coupling between myometrial cells during labor. The integration of these control mechanisms probably operates to ensure appropriate activation and maintenance of synchronous contractility of the myometrium and effective delivery. PMID- 3034461 TI - Synthetic ligaments. Current status. AB - Many techniques for ligamentous reconstruction have been developed in recent years. In the United States, injuries of the knee have been increasingly treated with innovative methods of surgical reconstruction, most of which have used normal structures. There are obvious theoretic advantages in using synthetic materials that might simplify surgery, spare normal tissues, and possibly facilitate stronger repairs. To these ends, several synthetic substances have been used experimentally and clinically. This is a brief summary of eight of the materials that have been or are being investigated in the United States. Some are no longer in use, others are currently being used in clinical trials. As of this writing, only the Gortex ligament has received a general device release from the Food and Drug Administration (FDA). PMID- 3034462 TI - Joint contractures in the hemophilias. AB - Hemophilic contracture is seen most commonly as an equinus deformity of the ankle, or at the knee or elbow in the form of a flexion deformity. The cause is either fibrosis following intramuscular hematoma, which may be complicated by a peripheral nerve palsy causing muscle imbalance, or is associated with chronic hemophilic arthropathy following recurrent hemarthroses. The introduction of home therapy is an important preventive measure, and treatment is primarily by physiotherapy, splintage, and corrective devices. The late or severe case may require surgical correction in the form of soft tissue procedures. Arthrodesis or total arthroplasty must be carried out with meticulous hemostasis and with replacement of the appropriate blood clotting factors. PMID- 3034463 TI - [Morphological and immunological studies of cerebrospinal fluid lymphocytes in two cases of Japanese encephalitis]. PMID- 3034464 TI - [Peripheral nerve involvement in chorea-acanthocytosis]. PMID- 3034465 TI - A parathyroid cyst with adenoma on thallium-201/technetium-99m subtraction imaging. AB - A case of a parathyroid cyst with adenoma was seen on Tl-201/Tc-99m subtraction imaging. The literature regarding parathyroid cysts and the subtraction technique for parathyroid adenoma imaging was reviewed. PMID- 3034466 TI - Radionuclide imaging of potassium iodide-induced sialadenitis. AB - The authors report a rare complication of nuclear bone imaging, iodide mumps, and describe its appearance on the subsequent radionuclide salivary gland study. PMID- 3034467 TI - Communicating gastric-subphrenic abscess proven by combined technetium-99m pertechnetate and sulfur colloid imaging. PMID- 3034468 TI - Age and the pharmacokinetics and pharmacodynamics of chronic enalapril treatment. AB - Enalapril clearance after single doses is reduced in the elderly. The influence of age on the pharmacokinetics and pharmacodynamics of chronic enalapril treatment was examined in six young (22 to 31 years) and six elderly (65 to 78 years) healthy subjects who took enalapril, 10 mg, daily for 8 days. The blood pressure fall was greater in the elderly even with chronic administration. Plasma angiotensin-converting enzyme inhibition was similar in both groups. Steady-state serum enalaprilat concentrations were achieved more slowly in the elderly subjects and were correspondingly higher for all subjects. Clearance/bioavailability and volume of distribution/bioavailability diminished with repeated administration. Repeated exposure also led to a reduction in sensitivity of plasma angiotensin-converting enzyme to the inhibitor. Prolonged inhibition probably induces synthesis of new angiotensin-converting enzyme. PMID- 3034469 TI - Inhibition of angiotensin-I response by cilazapril and its time course in normal volunteers. AB - Cilazapril is a new angiotensin-converting enzyme (ACE) inhibitor. In a double blind crossover study six normal male volunteers received single oral doses of cilazapril, 4 mg, captopril, 25 mg, enalapril, 10 mg, and placebo. The response of diastolic blood pressure to an intravenous infusion with increasing doses of angiotensin I (AT-I) (0.1 to 18 micrograms/min) was determined at control and up to 36 hours after oral drug intake. Additionally the response to AT-I was established before, during, and after cessation of a 15-day 2.5 mg/day cilazapril administration. The ACE inhibitors antagonized the AT-I effects and shifted the AT-I dose-effect curves rightward, whereas placebo was not effective. After single doses the effects of cilazapril and enalapril declined with a similar elimination half-life of approximately 4 hours; with captopril approximately 2 hours was observed. After multiple administration of cilazapril there was no evidence of cumulative effects. Cilazapril is an orally active ACE inhibitor that does not show pharmacodynamically relevant accumulation. PMID- 3034470 TI - Effect of chromocarb diethylamine on the permeability of the blood-brain barrier. AB - Intravenously injected collagenase, detectable in brain microvessels by immunological methods, partially degrades the constituents of the vascular wall and so increases the permeability of the blood-brain barrier (BBB). Intravenous administration of collagenase is a model for diseases in which the concentration of endogenous proteases is increased. Peroral treatment of rats with chromocarb diethylamine (CD) significantly reduced the degradation of the vascular wall by intravenous collagenase, as demonstrated by a lesser permeability increase of the BBB, a shorter recovery time, lower hydroxyproline levels in the cerebrospinal fluid and a lesser decrease of the collagen content of the brain capillary basal lamina. PMID- 3034471 TI - Humoral vasopressor agents in primary hypertension. AB - In essential hypertension and in the spontaneously hypertensive rat, there are indications for a yet unidentified vasopressor agent which may play an important role in pathogenesis. It has been demonstrated in the spontaneously hypertensive rat by parabiosis, cross-circulation, cross-transplantation of kidneys and by injection of hypertensive plasma fractions. In plasma from essential hypertensives, an agent sensitizing the vasculature to pressor hormones and a substance with direct vasopressor actions have been found. The relationship of the hypertensive factor to the natriuretic hormone is not clear. PMID- 3034472 TI - Benign neoplasms of the oral cavity. AB - A wide variety of benign tumors present in the oral cavity. These tumors are for the most part rare and are classified by the tissue of origin. Although benign oral cavity tumors are not life-threatening, they can result in extensive loss of soft tissue and/or bone. Furthermore, many patients are subject to the threat of recurrence, multiple surgical procedures, and the possibility of malignant degeneration. Because many tumors vary little clinically, an adequate biopsy specimen must be obtained for diagnosis. Radiographs are, in general, nondiagnostic. Collaboration with an experienced pathologist is necessary to determine the tumor's probable clinical behavior. Therapy, which is dictated by tumor type, is almost always surgical. PMID- 3034473 TI - Benign skin and soft-tissue tumors of the hand. AB - Tumors of the hand are relatively rare when compared to other body regions; the majority are benign in nature. Skin and soft-tissue benign tumors can be classified according to their tissue or origin. Proper diagnosis and differentiation of benign from premalignant or malignant lesions are the most important steps in the management of hand tumors. Surgical excision under adequate anesthesia with histologic diagnosis is the commonest and most accepted treatment. PMID- 3034474 TI - Detection of antibodies to anterior pituitary cell surface membrane with insulin dependent diabetes mellitus and adrenocorticotropic hormone deficiency. AB - Autoantibodies for anterior pituitary cell surface membrane (PitCSA) were assayed by immunofluorescence method using GH3 cells (rat GH and prolactin secreting cell) and AtT-20 cells (mouse adrenocorticotropic hormone secreting cell) as antigens. Out of 18 insulin dependent diabetic patients who were positive for antibodies to islet cell surface membrane (ICSA), 3 cases (16.7%) were positive for antibodies to GH3 cells and 12 cases (66.7%) were positive for antibodies to AtT-20 cells. Moreover, out of 18 insulin dependent diabetic patients who were negative for ICSA, 2 (11.1%) and 6 cases (33.3%) were positive for antibodies to GH3 cells and AtT-20 cells, respectively. Among 5 adrenocorticotropic hormone (ACTH) deficient patients, all of the sera were positive for antibodies to AtT-20 cells. These results suggested that PitCSA and ICSA have independent features, though both are closely related, and that PitCSA was one of the significant immunological markers often observed in the sera of the patients with insulin dependent diabetes mellitus (IDDM) and ACTH deficiency. PMID- 3034476 TI - Phyllodes tumour of the breast: response to radiotherapy. AB - A 47 year old woman presented with a rapidly growing recurrence in the chest wall following simple mastectomy for a malignant phyllodes tumour of the breast. Radical megavoltage irradiation led to histologically confirmed complete regression of her tumour. PMID- 3034475 TI - Cytotoxic islet cell autoantibodies in newly diagnosed insulin-dependent diabetes mellitus: lack of correlation to age, residual beta cell function, HLA antigens and Coxsackie B virus antibodies. AB - In a cross-sectional study comprising of 56 patients with newly diagnosed insulin dependent diabetes mellitus (IDDM) serum was examined for the presence of complement-dependent antibody mediated cytotoxicity (C'AMC) by an improved assay measuring the release of 51Cr from freshly isolated normal rat islet cells prelabeled with the isotope. In the presence of complement, 35 (63%) IDDM sera specifically mediated cytotoxicity against islet cells. The degree of cell specificity was tested using prelabeled exocrine cells, but this cell type did not reveal any differences between the lytic effects of IDDM and control serum. At the time of clinical onset of IDDM the age of the patients, fasting and stimulated C-peptide levels, insulin requirement, HLA antigens, antibodies to Coxsackie B1-6 viruses, diabetes heredity, and, surprisingly, islet cell surface antibodies (ICSA) were not associated with C'AMC. It is concluded that the mere presence of C'AMC against rat islet cells at the time of diagnosis of IDDM is not indicative of a low residual B cell function and HLA-linked susceptibility to diabetes does not necessarily include enhancement of humoral anti-islet cell autoimmunity. More elaborate methods for quantitative detection of C'AMC and ICSA (e.g. in terms of titres) together with autoantibody subclass determination are needed to reveal possible associations between C'AMC and the clinical characteristics of IDDM. PMID- 3034477 TI - High resolution proton nuclear magnetic resonance studies of human cerebrospinal fluid. AB - One- and two-dimensional (correlated shift spectroscopy) high resolution proton n.m.r. spectra of human cerebrospinal fluid (CSF) are reported. The merits of water suppression by freeze drying or irradiation, and spectral simplification by spin-echo methods, are discussed. Well-resolved resonances for a range of low molecular weight metabolites such as lactate, 3-D-hydroxybutyrate, alanine, acetate, citrate, glucose, valine and formate were observed. Resonances for glutamine were observed only from freeze dried samples. Concentrations determined by n.m.r. were in reasonable agreement with those from conventional methods. The n.m.r. spectra of CSF were related to the clinical conditions of the subjects. No resonances for citrate were present in spectra of CSF from subjects (three infants) with bacterial meningitis; high lactate and lowered glucose levels were observed. Strong resonances for glucose and glycine were observed for mildly diabetic subjects. Both the aromatic and the aliphatic regions of the CSF spectra from subjects suffering from liver failure contained distinctive features characteristic for hepatic coma: Intense resonances for lactate, alanine, valine, methionine, tyrosine, phenylalanine and histidine. In some cases guanine was also present, which does not appear to have been reported previously. The two dimensional spectrum suggested the presence of abnormally high levels of a number of endogenous metabolites. Such assignments were not possible using one dimensional spectra alone because of signal overlap. PMID- 3034478 TI - Blood gas measurement. PMID- 3034479 TI - [A cisplatin and etoposide combination in small cell anaplastic carcinoma of the lung]. PMID- 3034480 TI - [Phosphatidylserine and memory disorders in the aged]. PMID- 3034481 TI - Approaches to physical mapping of the human genome. PMID- 3034482 TI - Reduced recombination rate on chromosomes 21 that have undergone nondisjunction. PMID- 3034483 TI - Variability at the telomeres of the human X/Y pseudoautosomal region. PMID- 3034484 TI - Analysis of an X-autosome translocation responsible for X-linked muscular dystrophy. PMID- 3034485 TI - Repetitive human DNA sequences. PMID- 3034487 TI - Isolation and expression of cDNAs encoding human factor VII. PMID- 3034486 TI - Cloning of cDNA and genomic DNA for human von Willebrand factor. AB - The recent isolation of cDNA and genomic DNA clones for human vWF by ourselves and others has finally laid to rest the historical notion that factor VIII and vWF might have a precursor-product or other complex relationship. These two hemostatic activities are clearly present on two distinct proteins, each encoded by a separate gene. Whether ther is coordinate regulation of the factor VIII and vWF genes is still unknown. The structure and structure-function relationships of the vWF protein have been elucidated by many investigators, only some of whom can be cited in this short paper. Together, these molecular biology and protein chemistry studies have shown that vWAgII is the amino-terminal propeptide of vWF, explaining the proportional deficiency of these two plasma proteins in von Willebrand disease (type I). In addition, sites of proteolytic processing, glycosylation, and disulfide bond formation have been defined, and some of the binding functions of mature vWF have been identified with specific amino acid sequences. The protein has a highly repeated structure, suggesting a complex evolutionary history. The A domains of vWF appear to be homologous to complement factor B, and perhaps to component C2, although the biological meaning of this similarity is unknown. Genomic DNA clones have been isolated corresponding to approximately one third of the vWF gene on chromosome 12, and a fragment of the cDNA also hybridizes to uncharacterized sequences on chromosome 22. Preliminary studies in von Willebrand disease have revealed two patients with very large deletions in the vWF gene.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3034488 TI - Exploration of structure-function relationships in human factor VIII by site directed mutagenesis. PMID- 3034489 TI - Molecular and biological properties of human macrophage growth factor, CSF-1. PMID- 3034490 TI - Identification of DNA repair genes in the human genome. PMID- 3034491 TI - Structure of the activated c-raf-1 gene from human stomach cancer. PMID- 3034492 TI - Transfer of genes into human somatic cells using retrovirus vectors. PMID- 3034493 TI - Gene expression from a transcriptionally disabled retroviral vector. PMID- 3034495 TI - A milestone in human genetics. PMID- 3034494 TI - Gene transfer and expression in nonhuman primates using retroviral vectors. PMID- 3034497 TI - Cellular and molecular biology of lipoprotein metabolism: characterization of lipoprotein receptor-ligand interactions. PMID- 3034498 TI - Mapping of human chromosome 11: organization of genes within the Wilms' tumor region of the chromosome. PMID- 3034496 TI - Human steroid receptors and erbA proto-oncogene products: members of a new superfamily of enhancer binding proteins. PMID- 3034499 TI - The chromosome 11 gene map: genes for growth and development, Wilms' tumor deletions, and cancer chromosome breakpoints. AB - Human chromosome 11 is clearly a model autosome encoding genes and characteristics associated with both normal and abnormal growth and development, and several significant disorders. A fine-structure molecular, genetic, and physical map of this chromosome would add considerably to our knowledge of the organization and control of human genes and to an understanding of normal and abnormal human biology. PMID- 3034500 TI - The human myc gene family. PMID- 3034501 TI - Experimental infection of calves with strains of Bovid herpesvirus-4. AB - Fourteen calves were inoculated intranasally (i.n.) with the viral isolates as follows: 5 with 85/BH 16TV, 1 with 85/BH 17TV, 1 with 85/BH 18TV, 2 with 85/BH 231TN and 5 with 85/BH 232TN. Strain 85/BH 16TV was the only one which caused overt respiratory-like disease in all inoculated calves. Onset of the disease was observed after 7-8 days of incubation and was characterized by fever, depression, nasal discharge and coughing. Virus was isolated from the nasal swabbings of calves obtained from post-infection day (PID) 2-10. The other viral strains did not cause any sign of disease although virus was isolated regularly from the nasal swabbings of the inoculated calves. Virus was recovered from central nervous system tissues of calves that were infected with 85/BH 16TV or 85/BH 232TN strains and were killed on PID 4 or 8. Virus was also isolated from other tissues, such as lymph node, nasal mucosa (PID 8), or lung (PID 4). It was speculated that the nervous system could be one of the target areas of the virus of the naturally occurring infection by BHV-4. This might indicate a possible role of the nervous system (site of latency?) in the pathogenesis of BHV-4 as is the case in certain herpesviral infections of man and the lower animals. PMID- 3034502 TI - Reactivation of infectious bovine rhinotracheitis virus by transport. AB - Transport was studied as a cause of reactivation of infectious bovine rhinotracheitis virus (Bovine herpesvirus-1; BHV-1) in heifers vaccinated 2-6 months before transport, using a double dose of the thermosensitive (ts) vaccine strain (Tracherine). Eight out of 19 animals showed ts strain re-excretion over a period of 1-3 days, beginning, in 5 out of the 8 heifers, the day after transport. In 14 other heifers, only sera were examined by sero-neutralisation: only 1 out of these 14 animals showed a rise in BHV-1 neutralising antibodies. Transport can therefore be considered as a stimulus of BHV-1 reactivation. PMID- 3034503 TI - Induction and characterization of swine beta interferon. AB - Newcastle Disease Virus (NDV) strain "H" and Polyinosinic-Polycytidylic acid (Poly I:C) were used for interferon (IFN) induction in secondary pig kidney cells. A functional IFN system was detected and characterized. A wide similarity with the correspondent human and bovine systems was appreciated, with particular regard to the kinetics of synthesis. A glycosylated protein was essential for activity in bovine cells, but not in swine cells. Poly I:C proved to be a very weak inducer, even in conditions which promote IFN synthesis in other cell substrata. beta IFN from secondary pig kidney cells was very effective against Swine Vesicular Disease Virus (SVDV), whereas no activity was detected against porcine Rotavirus; Aujeszky's disease virus, BUK strain, proved to be of intermediate sensitivity. The results of these latter experiments are discussed, with regard to the cells used and to the IFN sensitivity of the tested viruses. PMID- 3034504 TI - Sarcolemmal ATPase activities of the rat heart ventricle--dependence on age and sodium ion. AB - Na-K-ATPase activity of the rat heart was similar throughout the postnatal growth when measured from crude unpurified fraction. Instead in the cardiac sarcolemmal fraction, isolated by hypotonic shock LiBr-treatment method, the activity was over two times higher in 10-day old neonates than in adult rats. The conflicting results are partly explained by different effects of the isolation procedure on neonatal and adult tissues. Na concentration for half-maximal activity of the Na K-ATPase was similar in neonates (7.0 mM) and adults (6.4 mM). Ca-ATPase activity was not affected by Na concentration (2-100 mM) in the two age-groups studied. PMID- 3034505 TI - Computed tomography of primary malignant fibrohistiocytoma of the lung. AB - A case of malignant fibrous histiocytoma of the lung is reported in pediatric patient-uncommon location as well as age group. PMID- 3034506 TI - Contact sensitivity to silicic acid in Unguentum Merck. PMID- 3034507 TI - HSV-1 latency: thymidine kinase requirement and the round-trip theory. AB - The present study used the rabbit iontophoresis model to examine the thymidine kinase (TK) requirement for HSV-1 latency, and test the round trip theory of latency with emergent phenotypic mutations of the HSV-1 TK negative (TK-) inoculating strain. The results demonstrated repeated induced ocular shedding of latent HSV-1 in 14-100% of rabbits. The TK phenotype of recovered tear film following spontaneous shedding and repeated iontophoresis induction was thymidine kinase negative in 90% of eyes. Sequential shedding of TK- virus from the same eye over time was demonstrated in 21% of eyes in 33% of rabbits. We conclude that TK is not an absolute requirement for the establishment or reactivation of latent HSV-1 in the rabbit model. The round trip theory of latency was not supported as TK+ isolates and syncytial variants of the TK- inoculating strain which were recovered at the ocular surface after the initial iontophoresis could not be demonstrated following subsequent trials of iontophoresis. PMID- 3034508 TI - Repair of traumatic splenic injuries by splenorrhaphy with polyglycolic acid mesh. PMID- 3034510 TI - Arenaviruses: biology and immunotherapy. PMID- 3034509 TI - Arenaviruses: genes, proteins, and expression. PMID- 3034511 TI - Neutralization of arenaviruses by antibody. PMID- 3034512 TI - Experimental studies of arenaviral hemorrhagic fevers. PMID- 3034513 TI - Argentine hemorrhagic fever. PMID- 3034514 TI - [Infection of the oral mucosa with human papilloma virus (HPV)]. PMID- 3034515 TI - Tumor imaging and targeting therapy for hepatocellular carcinoma. Preliminary results of experimental and clinical studies. PMID- 3034516 TI - N-ethyl-N'-nitro-N-nitrosoguanidine induced gastric carcinoma in wolfdogs--useful animal model for tracing gastric malignancy transformation. PMID- 3034517 TI - [Post-traumatic carpal tunnel syndrome]. AB - In the course of a ten months study on 508 patients with CTS a trauma as a possible causal factor was observed in 22 cases. In none of these cases did the other hand show normal electroneurographic parameters. The mean values of distal motor latency were among 5.5 ms (3.7-9.2 ms) on the injured side and 4.6 (3.5-6.7 ms) on the other. The relatively seldom occurrence of CTS in connection with a trauma requires therefore strict criteria for the evaluation of the casual context. A traumatic etiology can only be recognised if a close temporal relation exists (beginning of the symptomatology during immobilisation or after removal of the cast) or a special tendency to swelling or a dislocation as well as a clear difference in electroneurographic values of the two sides can be observed. Depending on the degree of this difference in latency one can maintain criteria for evaluating the question of causality or of transient or permanent deterioration. Finally in case of CTS the treatment should be given primary importance ahead of reimbursement for the injury. PMID- 3034519 TI - [Familial polyposis of colon (a study of 31 cases in 8 families)]. PMID- 3034518 TI - Regional assignment of five genes on human chromosome 19. AB - A human-mouse hybrid segregant HM76Dd40-6 with new characteristics was derived from the hybrid cell line HM76Dd containing human chromosome 19 as the only human chromosome. Three virus sensitivities located on human chromosome 19 (PVS, E11S and RDRC) were lost in HM76Dd40-6, while six other genes (C3, LDLR, EF2, GPI, PEPD and MANB) were retained. Cytogenetic analysis and in situ hybridization using human or mouse repeated sequences as probes showed that the region q13.1 qter of human chromosome 19 had been replaced by a fragment of mouse chromosome. Our results permit further regional assignment for the following five genes on human chromosome 19: GPI in the region cen-q12, MANB in p13.2-q12, E11S and RDRC in q13.1-qter, and EF2 in pter-q12. PMID- 3034520 TI - [The application of superselective external carotid arteriography and preoperative embolization]. PMID- 3034521 TI - Circadian rhythm of the angiotensin converting enzyme (ACE) activity in serum of healthy adult subjects. AB - Angiotensin converting enzyme (ACE) activity was measured in the peripheral serum of 10 healthy, diurnally-active and nocturnally-resting adult subjects (5 women and 5 men). Blood was drawn serially at 4-h intervals throughout a 24-h cycle with the subjects hospitalized and synchronized. They were not kept in recumbency. Data were evaluated by conventional statistical analysis and by single- and population mean- cosinor procedures. A low-amplitude circadian rhythm was statistically validated for the group. Acrophase was located in the afternoon, at about 1630. The recognition that ACE activity oscillates physiologically in the peripheral blood according to a circadian rhythm may serve to amplify the clinical use of ACE measurements. PMID- 3034523 TI - [Use of 99m TC scanning in diagnosing salivary gland tumors]. PMID- 3034522 TI - Differences in cortisol, aldosterone and testosterone responses to ACTH 1-17 administered at two different times of day. AB - The effects of 100 micrograms, i.m. of the analog ACTH 1-17 administered at 0800 and 1800 on the secretion of cortisol, aldosterone and testosterone have been studied in normal subjects: 8 male and 8 female. The group as a whole and the males had significantly greater absolute and percent increments in plasma cortisol after administration at 1800. In the females, there was only a greater percent increment in cortisol after the evening administration. The heptadecapeptide always significantly stimulated serum aldosterone, with no difference between the two times of administration. In the females, ACTH 1-17 significantly stimulated testosterone, with a more protracted secretion after the evening administration. In the males, there was always a significant testosterone decrease after the administration of the drug, with no difference between morning and evening. In conclusion, 100 micrograms i.m. of the analog ACTH 1-17 stimulates cortisol secretion more when given during the circadian nadir of plasma cortisol, but only in men. ACTH 1-17 increases testosterone in women and decreases it in men, whereas it seems to increase aldosterone secretion in both sexes. PMID- 3034524 TI - Remodelling of early axonal projections through the selective elimination of neurons and long axon collaterals. AB - Studies using neuroanatomical techniques have shown that the connections characteristic of the mature vertebrate brain are brought about by a considerable refinement of the projections initially established during development. The selective loss of neurons and long axon collaterals plays a major role in this remodeling process as illustrated in the development of the retina and cortex of the rat. In the retina, two-thirds of the initial population of ganglion cells (RGCs) die early. This loss serves to remove selectively RGCs that make erroneous axonal projections, including those which project to an incorrect target, to an inappropriate part of a correct target, or to the wrong side of the brain. Studies using the sodium channel blocker, tetrodotoxin, suggest that in rats the selective elimination of erroneously projecting RGCs is based, in part, on patterns of impulse activity. In the cortex a different mechanism is illustrated. All neocortical areas initially give rise to callosal and pyramidal tract axons but through a process of selective collateral elimination not involving cell death these projections assume the limited distributions seen in adult rats. Manipulations resulting in the maintenance of such long collaterals suggest that their removal is functionally and locally determined. In contrast to error elimination, this phenomenon of collateral elimination may be a developmental strategy for generating connectional diversity while limiting the amount of information required for the regional specification of the cortex. PMID- 3034526 TI - [Single fiber electromyography in myasthenia gravis]. PMID- 3034525 TI - Kainic acid: insights into excitatory mechanisms causing selective neuronal degeneration. AB - Kainic acid, an acidic pyrolidine isolated from the seaweed Digenea simplex, is the most potent of the commonly used exogenous excitotoxins. The neurotoxic threshold of kainic acid is nearly two magnitudes lower than that of the other receptor-specific agonists, N-methyl-D-aspartic acid and quisqualic acid. Neurophysiological and ligand-binding studies indicate that the neurotoxic action of kainic acid is mediated by a specific receptor which exhibits a remarkably broad phylogenetic distribution in the nervous system of vertebrates and invertebrates. The mechanism of neurotoxicity of kainic acid appears to be indirect and requires the functional integrity of excitatory afferents to vulnerable neurons. Consistent with the excitotoxin hypothesis, kainic acid depletes high-energy phosphates and glucose at sites of neurotoxic action; nevertheless, the proximate cause of neurotoxicity may involve increases in intraneuronal calcium levels and the activation of calcium-dependent proteases. Kainic acid neurotoxicity provides a useful animal model for selective neuronal vulnerability that may shed light on the pathophysiology of a number of neurodegenerative disorders, including Huntington's disease and temporal lobe epilepsy. PMID- 3034527 TI - [Analysis of the effectivity of treatment in 53 cases of small cell carcinoma of the lung]. PMID- 3034528 TI - [Ultrastructural study of 38 cases of pulmonary carcinoma]. PMID- 3034529 TI - The effect of smoking on myocardial metabolism. AB - The effect of passive smoking by the rabbits on the metabolism of myocardium was studied in three experimental models: following a single instance of smoking (for a period of thirty minutes); following a two-week smoking (twice a day for thirty minutes each); following an eight-week smoking (twice a day for thirty minutes each, i. e. for a period of 56 days. In isolated mitochondria of myocardium a decreased respiration, oxidative phosphorylation rates and cytochromoxidase activity were observed both after a single-instance smoking and prolonged smoking. The above disorders are closely related to the energy production in myocardium. Based on comparison of the above metabolic disorders with the changes in ultrastructure and myocardial function (described in the literature) the authors have come to the conclusion that the term "smoker's cardiomyopathy" which may develop in habitual smokers is well justified. This kind of cardiomyopathy may participate in the development of cardiac insufficiency even in the absence of atherosclerotic changes in the coronary arteries. PMID- 3034530 TI - Cytometry of breast carcinoma: significance of ploidy balance and proliferation index. AB - Image cytometry of DNA distribution in fine needle biopsies of breast carcinomas at first diagnosis was performed to see if there were significant differences in DNA histograms between patients having very different outcome but same tumor histological typing and similar therapy. Two groups of patients were considered retrospectively: the first (20 patients) with survival time shorter than 5 years and the second (20 patients) with survival time longer than 10 years. Seven benign tumors were used as controls. Ten ploidy classes were defined. The frequencies of cells in those classes were used as independent features in a supervised multivariate analysis. The advantages of this approach was pointed out with respect to the four-type classification of Auer. The scattering of DNA histograms within the feature space showed that a subgroup of patients with poor prognosis was clearly separated from a subgroup of patients with good prognosis but both long survival patients and short survival patients were scattered in between. In order to replace the multivariate classification of histograms by a simpler approach, two parameters were computed which explained most of the scattering in the feature space: the ploidy balance (difference between the percentages of euploid and aneuploid cells) and the proliferation index (percentage of cells between peaks). The scattergram of patients according to these parameters showed again that some DNA distributions were specific for either good or bad prognosis. But the separation was uncertain for seven short survival patients and six long-survival patients. For six patients, the DNA distributions were very similar between long and short survival times. Those patients thus could not be separated even by means of discriminant analysis. The main conclusion of this study was that, for a significant number of patients, the objective multivariate classification of tumors DNA profiles is of little assistance to the pathologist who has to give a prognosis for the one patient under consideration. PMID- 3034531 TI - Postjunctional alpha 2-adrenoceptors in pial arteries of anesthetized newborn pigs. AB - The purpose of this study was to characterize the nature of the postjunctional alpha-adrenoceptors (alpha 1 versus alpha 2) in the cerebral circulation of anesthetized newborn pigs. Diameters of pial arteries in anesthetized piglets, 1 6 days old, were examined using a 'closed' cranial window. We examined sympathetically mediated constriction of pial arteries in the presence and absence of alpha 1- (prazosin) or alpha 2- (yohimbine) adrenoceptor antagonists (1 mg/kg i.v.). Initial experiments demonstrated that prazosin and yohimbine attenuated constriction of pial arteries during topical administration to the pial surface of phenylephrine and clonidine, respectively. In one group, electrical stimulation of the ipsilateral superior cervical ganglion reduced pial arterial diameter from 176 +/- 26 (mean +/- SEM) to 153 +/- 24 microns, and this response was unaffected by prior administration of prazosin. In a second group, sympathetic nerve stimulation reduced pial arterial diameter from 175 +/- 23 to 148 +/- 19 microns, and this response was abolished by prior administration of yohimbine. After administration of yohimbine, pial arteries still constricted in response to topical phenylephrine and arterial hypertension. We conclude that sympathetic nerve stimulation in anesthetized newborn pigs constricts pial arteries via alpha 2-adrenoceptors. PMID- 3034532 TI - Effects of enalapril in insulin-dependent diabetic subjects with mild to moderate uncomplicated hypertension. AB - The antihypertensive efficacy of enalapril and its effects on the metabolism and kidney function were investigated in 11 insulin-dependent diabetic subjects with uncomplicated mild to moderate hypertension. During a short-term single-blind controlled trial, one daily dose of 20 or 40 mg enalapril significantly reduced both systolic and diastolic blood pressure. In the supine position, mean systolic blood pressure declined from 169 +/- 6 to 142 +/- 6 mmHg (P less than .01) and mean diastolic blood pressure from 101 +/- 1.5 to 85 +/- 2 mmHg (P less than .001). No changes in heart rate or postural hypotension were observed. During 1 yr of treatment, the antihypertensive efficacy of enalapril did not decline, and no clinical side effects were observed. Inhibition by enalapril of angiotensin converting enzyme did not modify daily insulin requirements, glycemic control, uricemia, or lipid metabolism; kalemia and the markers of diabetic nephropathy were not significantly altered. These results suggest that enalapril once daily should be used as the first step in the treatment of diabetic patients with mild to moderate hypertension. PMID- 3034533 TI - Factitious hypoglycemia due to administration of human synthetic insulin: new diagnostic challenge. PMID- 3034535 TI - Kaposi's sarcoma of the conjunctiva and CMV-retinitis in AIDS. AB - The number of AIDS patients is still increasing. In 30-50% of these patients ocular lesions occur, which are of diagnostic and prognostic significance. If the life-span of AIDS patients lengthens in the future, adequate treatment of the ocular conditions will become increasingly important. The two most important ocular manifestations AIDS are CMV-retinitis and Kaposi's sarcoma of the conjunctiva. DHPG, a new virustatic for human cytomegalovirus, appears promising as treatment for the severe CMV-retinitis, which leads rapidly to blindness. Two case histories illustrate the preliminary results obtained with DHPG treatment. Kaposi's sarcoma of the conjunctiva is relatively benign is AIDS and can be treated successfully by surgical excision, radiotherapy, cryotherapy or local injections of cytostatics. PMID- 3034536 TI - [Ultrasound-guided percutaneous fine-needle puncture cytology in the diagnosis of malignant tumors of the liver]. AB - Results of ultrasound guided percutaneous fine-needle puncture cytology in 142 cases of malignant tumors of the liver are reported. The positive cytology was noted in 118 (83.1%) (10 suspicious). The primary liver cancer comprised 99 cases. In 7 lesions, equal to or less than 3 cm in diameters, 6 were positive in cytology. In 20,3-5 cm in size, 18 were positive. In 15 false-negatives, 12 were larger than 5 cm in diameter. Among 43 cases of metastatic liver cancer, 34 showed positive cytology and 9 false-negative. Among 108 cases of benign hepatic diseases, in 49.1%, it was difficult to arrive at definitive diagnosis by ultrasonography only, but in 97.2% malignancy was excluded by ultrasound guided fine-needle puncture cytology. The suspicious false-positive result occurred only in 3 cases. In this series, there were 250 cases of malignant tumors and benign diseases. The overall accurate diagnostic rate was 89.2%. All the patients had been followed for more than 6 months. The differential diagnosis between malignant and benign tumors, causes of misdiagnosis and complications are discussed. PMID- 3034537 TI - [CMC chemotherapy regimen combined with surgery of small cell lung cancer (SCLC)]. AB - From Dec. 1982 to Oct. 1984, 35 patients with SCLC proved by pathology or cytology, were treated by cyclophosphamide + methotrexate + CCNU (CMC) regimen combined with surgery in our hospital. All the patients received chemotherapy for more than 2 courses and the overall response rate was 85.7%, complete remission (CR) rate was 14.3%. Toxic reactions were tolerable to the patients. Treatment result was better in SCLC with localized than extensive disease. Operation was done for 9 out of 21 patients with localized lesions which had responded to chemotherapy. Of them, 1 died of postoperative complication, 2 were lost in follow-up and the rest 6 were disease-free for 8-32 months with a median survival time of 19 months. The 1 year survival rate was 75%. The results indicate that in limited disease of SCLC, successful chemotherapy combined with surgery can prolong the survival time. For patients with an limited disease which has given a CR, surgical resection should be strived for. PMID- 3034534 TI - Genomic clones encoding chicken myosin heavy-chain genes. AB - A chicken genomic library was screened with a cDNA probe containing the 3' coding and noncoding portions of quail fast-twitch skeletal muscle myosin heavy chain (MHC). This probe hybridized to seven to nine bands on Southern blots of chicken genomic DNA, and 17 clones that hybridized to this probe were obtained from the genomic library. Partial restriction maps were constructed and probable orientation of transcription was determined for each of the 17 clones. These maps indicate the presence of at least 14 unique MHC genes or pseudogenes. Dot-blot hybridization analysis using DNA complementary to RNA from a variety of chicken tissues demonstrated that these genes are all related to the gene for sarcomeric MHC, and permitted tentative assignment of the tissue of expression for several of the MHC isoforms. To substantiate further the dot-blot data, a subclone of one of the genes (4b1), which showed significant homology with adult breast muscle RNA but which also showed weaker hybridization to RNA from other tissues, was sequenced. The sequence data verified that the clone contains a portion of a MHC gene, that it contains both 3' coding and noncoding regions, and that its predicted amino acid sequence is identical (with 96% nucleotide homology) to that of the 75-bp quail fast MHC cDNA clone published by Hastings and Emerson (1982). Thus, clone 4b1 contains a portion of one of the genes that is expressed in adult chicken breast skeletal muscle tissue. PMID- 3034538 TI - [Randomized clinical trial of cis-platinum diamminedichloride (PDD) in the treatment of hepatocellular carcinoma (HCC)]. AB - From Oct. 1982 to Apr. 1985, 82 patients with HCC proven by pathology were treated in our hospital. 43 treated by hepatic arterial perfusion, were randomized into PDD group: PDD 10 mg per day X 10, every 3 weeks; control group: fluorouracil (5-Fu) 250 mg per day X 4, every week and thio-tepa (TSPA) 10 mg, twice a week. The other 39 treated by intravenous chemotherapy, were also randomized into PDD group: PDD 20 mg per day X 5, every 3 weeks; control group: 5 Fu 500 mg and TSPA 10 mg, twice a week. The objective response rates were 31.8% (7/22) in PDD group and 23.8% (5/21) in control group by hepatic arterial perfusion, and 20.0% (4/20) in the former and 0% (0/19) in the latter who were treated intravenously. The median survivals were 8 months for all the patients receiving hepatic arterial perfusion, and 6 and 5 months for the intravenous PDD and its control group, respectively. The side effects and kidney toxicity of PDD were tolerable to the patients. It is observed that PDD is better than 5-Fu and TSPA in the treatment of HCC. PMID- 3034539 TI - [Resection and reconstruction of the carina]. AB - A case of recurrent adenoid cystic carcinoma after right middle lobectomy is reported. A right lower lobectomy with resection of carina was performed as the lesion involved the right main bronchus, the carina and the orifice of right lower lobe. The carina was reconstructed by the anastomosis of right upper lobe bronchus to the left main bronchus. The postoperative course was uneventful. After two months, the stitch granuloma at the site of anastomosis developed and it was satisfactorily managed by local palliative measures. No recurrence has been found during a follow-up of 1.5 years. PMID- 3034540 TI - [Bireplicon vector plasmids for the cyanobacterium Synechocystis 6803 and Escherichia coli]. PMID- 3034541 TI - [Cytomegalovirus hyperimmunoglobulin prophylaxis following kidney transplantation. Results of a prospective randomized study]. AB - The effectiveness of preventing cytomegalovirus (CMV) infections by administering CMV hyperimmunoglobulin was evaluated in a prospective randomized trial. The patients in the treatment group (n = 50) had intravenous infusions of 2 ml/kg bodyweight of CMV-Polyglobin at three-week intervals, up to day 105 after kidney transplantation. The 50 patients in the control group received no infusions. There was no significant difference between the treatment and control groups in transplant survival or patients survival rates. But the number of symptomatic CMV infections was higher in the control (n = 11) than the treatment group (n = 5). There were also significantly fewer symptomatic herpes-simplex infections in the treatment (n = 6) than in the control group (n = 25). It is concluded from these results that prophylaxis with CMV hyperimmunoglobulin should be undertaken either selectively or for shorter periods than those chosen for the reported trial. PMID- 3034542 TI - [Conservative cholelitholysis in the calculus-congested gallbladder]. AB - Daily administration of 500 mg ursodeoxycholic acid and chenodeoxycholic acid (divided into two doses, morning and evening, corresponding to 8.5 mg per kg body weight and substance), all concrements in the gall-bladder were dissolved over a period of 14 months in a 60-year-old man for whom surgical treatment was contra indicated because of severe coronary heart disease. This case illustrates that, in the face of a high operative risk, the attempt to dissolve stones in the gall bladder by drugs is justified. PMID- 3034544 TI - [Current status of the tumor risk from oral contraception. The statement of the Standing Committee on Hormone Toxicology of the German Society for Endocrinology]. PMID- 3034545 TI - [Resistance against new gyrase inhibitors--1985/86]. PMID- 3034546 TI - Malignant diseases of childhood seen at the University of Nigeria Teaching Hospital (UNTH), Enugu, Nigeria. PMID- 3034543 TI - [Epidemiological and clinical aspects of focal nodular hyperplasia of the liver. An assessment of 886 cases]. PMID- 3034547 TI - [Joint amplification of c-myc and c-Ha-ras oncogenes in human breast and thyroid cancer cells]. AB - Both c-myc and c-Ha-ras 1 oncogenes amplification and enhanced expression were revealed in some human mammary and thyroid carcinomas by the molecular genetic analysis. Amplification proves to be a second possible molecular mechanism of the ras family protooncogene activation besides a point mutation. The coexistence of c-myc and c-Ha-ras 1 in some human primary carcinomas suggests a multistep process of carcinogenesis. PMID- 3034548 TI - [Intensity of the synthesis of RNA precursors in blood lymphocytes of cattle infected with leukemia virus]. AB - The peripheral blood lymphocytes of the cattle with lymphatic leukemia experimentally induced by BLV and of the animals from the same group with persistent lymphocytosis show the lower level of the pre-RNA-synthetic activity per cell when compared with the lymphocytes of healthy cattle (as evaluated by autoradiography with 3H-uridine as a precursor). At the same time the RNA synthetic activity in the population as a whole increases as a result of growth of the RNA synthesizing cells (a labelling index), which is confirmed by enhancement of DNA-dependent RNA-polymerase activities and 3H-uridine in vitro incorporation into lymphocytes revealed by the liquid scintillation counting method. PMID- 3034550 TI - F wave measurements: a comparison of various recording techniques in health and peripheral nerve disease. PMID- 3034549 TI - [Complex evaluation of the proliferative activity of tumor cells in different histological variants of lung cancer]. AB - Mitotic regime, labelling index and DNA distribution in malignant lung tumours of different histological types (47 cases) were investigated. The most pronounced proliferative activity in combination with high incidence of three group metaphases and multipolar mitoses of bi- and polyclonal cases of undifferentiated cancer and poorly differentiated squamous cell carcinoma were established. Well differentiated tumours were monoclonal and usually had a low labelling index. The high incidence of c-mitoses in well-differentiated squamous cell carcinomas is, probably, an evidence for a more pronounced damage of their cell multiplication. PMID- 3034551 TI - Normal H reflexes and denervation in the low leg musculature: comparison of EMG with computed tomography, myelography and operation in patients with radiculopathy. PMID- 3034552 TI - Investigations on the peripheral motor neurone in subjects with alcohol dependence syndrome. PMID- 3034553 TI - [Current hypotheses on the mechanism of action of lithium. II. Lithium and membranes; from the receptor to cyclic nucleotide by the way of membrane phospholipids]. AB - This article details how lithium affects the membrane. Lithium would counter variations in receptor sensitivity. This hypothesis has been taken up by many authors working with different models. The practical implications of such an effect remain to be defined. Also, lithium would act on the metabolism of cyclic nucleotides, principally cyclic AMP. The enzyme adenylate-cyclase, which synthesizes the nucleotide, might be one of the main targets of its therapeutic action. Between the receptors at the exterior of the membrane and the second messengers inside the cell, the mechanisms of information transfer across the membrane (or transductor system) would in turn be affected by the cation. More precisely, by interfering with the metabolism of phospholipids, essential components of this system, lithium would modify the usual functioning of the transductor. All these observations suggest a global stabilizing effect on the membrane. PMID- 3034554 TI - [Postmortem studies of the brain of suicide victims]. AB - The authors present a bibliography concerning post mortem studies in depression and suicide victims. They stress the difficulties of this trans-disciplinary approach, joining clinical, toxicological, anatomopathological, and biological points of view. Concerning indolamines, catecholamines, receptors studies, an analysis of the results shows major methodological differences. This makes any comparison between these studies impossible. Recommendations are made for an agreement between the groups leading to an uniformity of methodologies. PMID- 3034555 TI - Effects of ovine corticotropin-releasing hormone and FK 33-824 (met-enkephalin analogue) on the secretions of proopiomelanocortin-derived N-terminal peptide, beta-lipotropin, beta-endorphin and adrenocorticotropin in patients with Addison's disease. AB - The responses of plasma immunoreactive (IR) proopiomelanocortin (POMC)-derived N terminal peptide (Nt), IR-beta-endorphin (Ep), IR-beta-lipotropin (LPH) and IR ACTH levels to ovine corticotropin-releasing hormone (CRF) and FK 33-824 (Met Enkephalin analogue) were studied in nine patients with Addison's disease. The basal plasma levels (mean +/- SE) of IR-Nt, IR-Ep, IR-LPH and IR-ACTH were significantly higher in patients with Addison's disease (4459 +/- 975 pg/ml, 132 +/- 25 pg/ml, 4425 +/- 1030 pg/ml, 553 +/- 89 pg/ml, respectively) than in the normal controls (202 +/- 38 pg/ml, 7 +/- 2 pg/ml, 101 +/- 18 pfi/ml, 53 +/- 16 pg/ml, respectively). Ovine CRF produced rapid and concomitant increases in plasma levels of IR-Nt, IR-Ep, IR-LPH and IR-ACTH. Ep and ACTH levels reached a peak at 30 min. On the other hand, Nt and LPH levels reached a peak at 60 min and these levels gradually decreased up to 120 min. The molar concentrations of these IR-peptides in plasma were changed in close parallel fashion to one another. FK 33-824 produced a pronounced and concomitant fall in IR-Nt, IR-EP, IR-LPH, and IR ACTH levels. These results support the theory that Nt, Ep, LPH and ACTH are produced simultaneously from POMC as a common precursor in the pituitary gland and are secreted concomitantly under various conditions such as stimulation by CRF and inhibition by FK 33-824 in patients with Addison's disease. PMID- 3034556 TI - Two adult familial cases of selective hypoaldosteronism due to insufficiency of conversion of corticosterone to aldosterone. AB - A 57-year-old woman (case 1) and her daughter aged 29 (case 2) with hyperkalemia exhibited subnormal plasma aldosterone (ALD) in the face of elevated plasma renin activity. Their physical findings were normal. Their arterial blood gas analysis showed that metabolic acidosis and renal function of these cases were slightly impaired. Urinary 17-OHCS and 17-KS excretions in these cases were normal. Baseline levels of corticosterone (B) and 18-hydroxycorticosterone (18-OH-B) were clearly elevated. Plasma deoxycorticosterone (DOC), B and 18-OH-B as well as cortisol remarkable increased after ACTH injection, but the increase in plasma ALD was very small. Angiotensin II infusion in case 1 resulted in a clear rise in plasma 18-OH-B but in slight depletion of B, and no increase in ALD. 9-alpha fludrocortisone acetate treatment was performed in case 1. Serum potassium was normalized and blood pressure elevated from 82/52 to 120/78 mmHg. Arterial blood gas analysis was corrected. We concluded that these two cases with subnormal plasma ALD and hyperreninemia may exist as a congenital and familial abnormality of the final step of aldosterone boisynthesis due to the impairment of the conversion of B to ALD. PMID- 3034557 TI - Effects of (D-ala2, met5)-enkephalinamide and naloxone on ACTH and corticosterone secretion. AB - An intravenous administration of (D-ala2, met5)-enkephalinamide (DALA) caused a significant elevation of plasma ACTH and corticosterone at 10 to 20 min after injection in unanesthetized freely moving rats. An intraperitoneal administration of cyproheptadine tended to reduce plasma ACTH and corticosterone levels at 60 min after injection, but it did not attenuate the DALA-induced ACTH and corticosterone elevation. A large dose of naloxone (1-10 mg/kg body weight) caused a significant elevation in plasma corticosterone, but naloxone at 10 mg/kg body weight reduced the basal ACTH level and DALA-induced ACTH elevation. When both DALA and naloxone were injected, the steroidogenic effect was attenuated. Neither DALA nor naloxone affected the basal ACTH release and CRF-induced ACTH stimulation in rat anterior pituitary cell cultures. These results suggest that DALA acts at the extra-hypophyseal level to stimulate ACTH and corticosterone and that the naloxone stimulatory effect on steroidogenesis acts on the adrenal gland or is mediated by stimulating corticosterone stimulating factors other than ACTH. PMID- 3034558 TI - Evidence for decreased activity of guanine nucleotide binding protein in adenylate cyclase of cell membranes in human ACTH-unresponsive adrenocortical carcinoma. AB - The present investigation was performed in order to study the properties of abnormal membrane function related to ACTH receptor-adenylate cyclase system interaction in human ACTH-unresponsive adrenocortical cancer. Two tissues of adrenocortical cancer obtained from a patient with Cushing's syndrome (CS) and a case presenting no abnormal endocrinological findings (NF) were used for in vitro studies, comparing with three normal adrenal tissues. The addition of ACTH alone and ACTH plus 10(-6) M GppNHp did not enhance the adenylate cyclase (AC) activity in the CS and NF tissues. Relative insensitivity of AC to GTP, GppNHp, and cholera toxin was observed for the NF tissue, while the rate of response to GppNHp for the CS tissue which also showed relative insensitivity to GTP and cholera toxin was similar to that for the normal tissues. Forskolin which is reported to directly activate the catalytic unit of the AC complex increased the AC activity of both CS and NF tissues as well as that of the normal tissues. Therefore, the function of the catalytic unit itself may be rather well preserved in these tumor tissues. These results suggest that the lack of ACTH receptor at the cell membrane surface might be responsible for ACTH-unresponsiveness in the CS tissue, although an accelerated degradation of GTP could contribute to decreased activity of GTP-binding protein.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3034559 TI - Response of anterior pituitary cells to culture media. AB - We examined the ability of four commonly used culture media to support prolactin (PRL), growth hormone (GH), and adrenocorticotropic hormone (ACTH) release, as well as the inhibitory PRL response to dopamine. After a week of primary culture, rat anterior pituitary cells from both genders were studied over a 4 hour period. Whereas ACTH secretion was similar across the various media, PRL and GH release were lessened with M199 and F10, respectively. Dopamine inhibited PRL release under all media conditions which was inconsistant with the reported lack of a dopamine effect with RPMI-1640 medium. These data confirm the postulate that media culture conditions can determine the degree of expression of constituitive phenotypes in anterior pituitary cells. PMID- 3034561 TI - Direct power-frequency electric field effects on mammalian endocrine tissue. AB - A number of studies have investigated the in vivo biological effects of power frequency electric fields (EF). Direct effects of EF on mammalian tissues, however, have rarely been reported. We now report that a 60-Hz EF can directly enhance the steroidogenic response of superfused rat adrenocortical tissue. The EF did not influence basal steroidogenic activity, however, the corticosterone response to 10 mU of ACTH was almost doubled by an unperturbed 1000 kV/m EF during the initial 2 hr of exposure and was enhanced fourfold by 5.5 to 7 hr of exposure with a 10 kV/m EF. Other EF intensities (e.g., 5 and 100 kV/m) were without effect at these times. Turning the 1000 kV/m EF on and off at 30-min intervals did not influence the initial enhanced steroidogenic response but did cause an additional two- to threefold elevation in the response following 5.5-7 hr of exposure. It is not clear what EF exposure parameters or mechanisms were primarily responsible for these bioeffects, but it appears that direct exposure of mammalian endocrine tissue to a 60-Hz EF is capable of significantly influencing important cellular processes. PMID- 3034560 TI - Long-term increase in striatal thyrotropin-releasing hormone receptor binding caused by amygdaloid kindling. AB - Our previous finding that intracerebroventricular (i.c.v.) administration of both thyrotropin-releasing hormone (TRH) and its analogue, gamma-butyrolactone-gamma carbonyl-L-histidyl-L-prolinamide citrate (DN-1417), suppressed seizure development of amygdaloid (AM) kindling and kindled AM seizures leads to a new hypothesis that endogenous TRH may be an antiepileptic substance in the brain. In this study, we examined postictal chronological changes in both immunoreactive TRH (IR-TRH) and TRH receptor binding activity in discrete brain regions of AM kindled rats to study the relationship of the brain TRH system to kindling induced seizure susceptibility. AM-kindled rats were decapitated 30 min, 24 h, 48 h, 7 days, and 21 days after the last kindled convulsion. IR-TRH increased markedly in the AM/pyriform cortex and hippocampus 24 and 48 h after the last convulsion, and returned to the control (unstimulated, sham-operated) value within 3 weeks after the convulsions ended. In contrast, a significant increase in the striatal TRH binding sites was evident 24 h after the cessation of convulsions which lasted 21 days. A lasting change in the striatal TRH neural system may be related to kindling-induced seizure susceptibility. PMID- 3034562 TI - Effects of in vitro asbestos exposure on natural killer and antibody-dependent cellular cytotoxicity. AB - Human peripheral blood lymphocytes (PBL) were exposed in vitro to asbestos fibers. Antibody-dependent cellular cytotoxicity (ADCC) activity and natural killer (NK) activity were examined by a chromium-51 release assay. There was a statistically significant enhancement of ADCC and NK activity by chrysotile and crocidolite fibers when cultured together with PBL for a period of 42 hr in medium containing a concentration of at least 2.5% fetal calf serum (P less than 0.05). Isolation of large granular lymphocytes to measure NK activity, however, showed the opposite effect when exposed to asbestos fibers (P less than 0.05). Our results indicate that asbestos fibers can directly affect lymphoid cytotoxic responses in vitro and may provide clues to immunopathogenic mechanisms for the occurrence of neoplasms in vivo. PMID- 3034563 TI - Crocidolite-induced lipid peroxidation in rat lung microsomes. I. Role of different ions. AB - Lipid peroxidation is increased by ferrous or ferric ions in rat lung microsomes both from rats pretreated with 3-methylcholanthrene and from untreated controls. This increase was dependent on the concentration of these ions in the reaction mixture. Crocidolite alone increased peroxidation in microsomal fractions. However, addition of ferric or ferrous ions with the crocidolite did not give a greater increase in the amount of peroxidation in microsomes. Chelation with EDTA of iron, whether originally present as free ions in the solution or attached to crocidolite, prevented lipid peroxidation. NADPH alone, when added to the microsomal fractions, did not produce any significant effect. However, when added concomitantly with crocidolite fibers, NADPH reduced the effect of crocidolite on lipid peroxidation. Magnesium, manganese, and calcium ions did not produce any significant effect on lipid peroxidation in the presence or absence of crocidolite. A reduced pH enhanced the rate of lipid peroxidation in line with increased solubility of iron salts at these pH values. All the above observations, taken together, lead to the conclusion that it is the iron in the crocidolite that is responsible for the latter's ability to enhance lipid peroxidation. PMID- 3034564 TI - A predictive model for determining asbestos concentrations for fibers less than five micrometers in length. AB - The controversy of whether small asbestos fibers are biologically significant has not been resolved. The present standard method for evaluating asbestos fiber concentrations in workroom air excludes fibers less than 5 micron long even though it has been shown that small fiber concentrations dominate in a dust cloud. This research project was conducted to develop a mathematical model whereby one could predict small (less than 5 micron length) asbestos fiber concentration based on the fiber count concentration determined by phase contrast microscope analysis. Dry chrysotile asbestos was aerosolized into a chamber and sampled by membrane filtration. Segments from each filter were analyzed by both the NIOSH technique using phase contrast microscopy (PCM) and scanning electron microscopy (SEM) at 2000 X for fiber concentrations. A linear relationship was found to exist between the natural logarithm of the SEM-determined concentration and the natural logarithm of the PCM-determined concentration (r = 0.852). Using these data, a mathematical model was developed to predict SEM concentrations based on PCM counts. This model may have application in retrospective epidemiological studies for estimating small fiber exposure levels to determine if small fibers play a role in disease production. The greatest utility would be in those retrospective studies where the only exposure information available is based on PCM counts. PMID- 3034565 TI - Methylmercury chloride-induced and antagonist-reverted succinic dehydrogenase changes in the brain and trigeminal ganglia of the rat. AB - The effect of methylmercury chloride (MMC) toxicity and its antagonism by chelating agents (N-acetyl-DL-homocysteine thiolactone and 2,3-dimercaptosuccinic acid) on succinic dehydrogenase (SDH) activity of fore-, mid-, and hindbrain and trigeminal ganglia of rats is reported in this study. A dose of 10 mg MMC/kg body weight was injected subcutaneously for 2, 7, and 15 days. The chelating agents were also injected subcutaneously in two separate groups of MMC-treated animals except in group 3. In the latter case MMC was injected in two groups of rats for 7 days, and thereafter antagonists were administered daily (40 mg/kg) for 1 week. The result shows inhibition of the enzyme with MMC in all groups and its restoration by N-acetyl-DL-homocysteine thiolactone; 2,3-dimercaptosuccinic acid, however, further reduced the enzyme level. The significance of inhibition of the enzyme in relation to tissue respiration and ATP production is discussed and the capacities of antagonists in the restoration of the SDH level are also analyzed. PMID- 3034566 TI - Inhibition of hepatic 17 beta- and 3 alpha-hydroxysteroid dehydrogenases by antiinflammatory drugs and nonsteroidal estrogens. AB - Antiinflammatory agents and estrogens have been tested as inhibitors of two isozymes of guinea pig liver testosterone 17 beta-dehydrogenase (NADP) 1.1.1.64) and rat liver 3 alpha-hydroxysteroid dehydrogenase (EC 1.1.1.50). Antiinflammatory steroids and estradiols were highly inhibitory to 3 alpha hydroxysteroid dehydrogenase and one isozyme of testosterone 17 beta dehydrogenase, respectively, but nonsteroidal antiinflammatory agents and nonsteroidal estrogens such as hexestrol, dienstrol, diethylstilbestrol and zearalenone showed potent inhibitions on all the enzymes. Although the inhibitory potency of indomethacin for one isozymes of testosterone 17 beta-dehydrogenase and 3 alpha-hydroxysteroid dehydrogenase decreased with changing pH from 9.7 to 7.0, that of the nonsteroidal estrogens for all the enzymes was little affected by pH. No additive effect in double inhibitor experiments with indomethacin and the nonsteroidal estrogens was observed, and the compounds were all competitive inhibitors with respect to steroidal substrate. The results suggest that there is a very similar region in substrate binding sites of the enzymes. PMID- 3034567 TI - Cloning and expression of the cDNA coding for a human lymphocyte IgE receptor. AB - Low-affinity receptors (Fc epsilon R) and secreted factors (IgE-BF) which bind to immunoglobulins of the IgE isotype play a key role in the regulation of human IgE synthesis. We report here the cloning of a cDNA coding for the Fc epsilon R of the human B-lymphoblast cell line RPMI 8866. The nucleotide sequence of this cDNA predicts a polypeptide with 321 amino acids and a mol. wt of 36,281 daltons. A functional Fc epsilon R capable of binding IgE was expressed in Chinese hamster ovary cells after stable transformation with the cDNA which had been cloned into a mammalian expression vector. Amino acid sequence analysis of IgE-BF purified from RPMI 8866 cells revealed an amino-terminal sequence of 19 residues which coincides with the predicted amino acid sequence of the Fc epsilon R, starting at residues 148 and 150. A computer search with the translated amino acid sequence of the Fc epsilon R revealed a domain of 120 amino acids having striking homology to the human asialoglycoprotein receptors. PMID- 3034568 TI - Oncogene expression during progression of mouse mammary tumor cells; activity of a proviral enhancer and the resulting expression of int-2 is influenced by the state of differentiation. AB - The RAC cell lines are derived from a mammary tumor induced by the mouse mammary tumor virus (MMTV). Polygonal cells have retained many characteristics of epithelial cells and induce adenocarcinomas; cuboidal cells are poorly tumorigenic; and elongated cells produce highly malignant sarcoma-like tumors. MMTV proviral integrations, one of which has cis-activated the int-2 gene, demonstrated that the lines are clonal descendents from a single tumor cell. Polygonal cells have a high constitutive expression of MMTV and contain int-2 RNA. In contrast, the cuboidal and the elongated cells show no detectable expression of int-2, even in the presence of glucocorticoid hormone. Transcription of MMTV in these cells is also low but can be stimulated by dexamethasone, albeit to levels lower than in polygonal cells. Thus, expression of int-2 seems to be caused by an enhancing activity on the MMTV provirus which is not dependent on steroid hormone and is specific for mammary tumor cells with epithelial characteristics. Progression in this cell system does not require sustained expression of int-2. PMID- 3034569 TI - Transfection of the int-1 mammary oncogene in cuboidal RAC mammary cell line results in morphological transformation and tumorigenicity. AB - The int-1 gene is often activated by proviral insertion in mouse mammary tumors. Direct evidence for the normal function of this gene and its role in tumorigenesis has therefore been lacking. To examine possible biological effects of int-1 activation in in vitro cell systems, we have constructed recombinant molecules of genomic int-1 DNA, transcriptionally activated by retroviral promoters. Transfection of these constructs into cuboidal RAC311C mammary cells leads to morphological transformation of the cells and rapid tumorigenicity. PMID- 3034570 TI - Expression of the human papillomavirus type 18 E7 gene by a cassette-vector system for the transcription and translation of open reading frames in eukaryotic cells. AB - We have constructed and functionally tested a cassette-vector-system for the transcription and translation of open reading frames (ORFs) in cells of higher eukaryotes. The vectors are derived from the plasmid pBR322 and can be selected and amplified in Escherichia coli. Alternative eukaryotic promoters can be inserted between the restriction sites SphI and KpnI, translation initiation motifs between KpnI and BglII, linkers for the adjustment of the translation reading frame and the insertion of genes or gene segments between BglII and HindIII, followed by a HindIII-EcoRI segment with splicing and polyadenylation signals derived from SV40. A prototype vector system, pORFEX11, 12 and 13, contains the strong cytomegalovirus immediately early promoter and a 10-bp motif of the SV40 T-antigen translation start. Polylinkers derived from pUC18 permit the insertion of ATG-less ORFs downstream from the ATG of the vector. Either of the three alternative polylinkers adjusts the appropriate translation frame. A similar construct contains the regulatable promoter of the Drosophila heat shock gene 70. We inserted genes or gene segments, that code for the bacterial chloramphenicol acetyltransferase, the bacterial gene conferring resistance against hygromycin, and the ORF E7 of the human papillomavirus type 18 into these vectors. After transfection of mouse L fibroblasts, all proteins and functions were expressed in accordance with the prediction. In transiently transfected L cells, the E7 protein expressed from pORFEX12 constitutes approximately 2.0% of total cell protein. This E7 protein could be localized by immunocytochemistry as a cytoplasmic component. PMID- 3034571 TI - Identification of early proteins of the human papilloma viruses type 16 (HPV 16) and type 18 (HPV 18) in cervical carcinoma cells. AB - We have sequenced 1730 bp of human papilloma virus type 18 (HPV 18) DNA containing the open reading frames (ORF) E6, E7, the N-terminal part of E1 and, additionally, 120 bp of the N-terminal part of L1. Based on these sequencing data, together with the human papilloma virus type 16 (HPV 16) DNA sequence published recently, we identified and cloned the ORF E6, E7, E1 and L1 of HPV 18 and the ORF E6, E7, E1, E4, E5, L2 and L1 of HPV 16 into prokaryotic expression vectors. The expression system used provides fusions to the N-terminal part of the MS2 polymerase gene controlled by the heat-inducible lambda PL promoter. Using the purified fusion proteins as immunogens we raised antisera against the proteins encoded by the ORF E6, E7 and E1 of HPV 18 as well as those encoded by the ORF E6, E7, E4 and L1 of HPV 16. By Western blot analysis we could show that the E7 gene product is the most abundant protein in cell lines containing HPV 16 or HPV 18 DNA. It is a cytoplasmic protein of 15 kd in the SiHa and the CaSki cell lines which contain HPV 16 DNA, and 12 kd in the HeLa, the C4-1 and the SW756 cell lines which contain HPV 18 DNA. These results were confirmed by in vitro translation of hybrid-selected HPV 16 and HPV 18 specific poly(A)+ RNA from SiHa, CaSki and HeLa cells. Additionally, these experiments led to the identification of an 11-kd E6 and a 10-kd E4 protein in the CaSki cell line as well as a 70-kd E1 protein in HeLa cells. PMID- 3034572 TI - Trans-activation of an upstream early gene promoter of bovine papilloma virus-1 by a product of the viral E2 gene. AB - The approximately 1000 nucleotide long upstream regulatory region (URR) of bovine papilloma virus-1 (BPV-1) contains a cis element which responds to trans activation by a diffusible factor encoded in the viral E2 open reading frame (ORF). A series of URR DNA fragments have been linked to two heterologous genes, bacterial chloramphenicol acetyl transferase (cat) or herpes simplex virus-1 thymidine kinase (tk), and tested in transient transfection assays for transcription initiating at the authentic upstream early viral promoter, P89. Transcriptional activity of the P89 promoter was greatly elevated in the presence of the E2 trans-activator gene product. The E2-responsive cis element (E2R) of P89 has been mapped to sequences -277 to -131 nucleotides upstream from the transcription start site (BPV nucleotide 89). The E2R element functioned as a strong transcriptional enhancer in cis with the SV40 early or the tk promoter in the presence, but not in the absence, of the E2 gene product. However, several heterologous promoters which lack sequences related to the E2R element were also trans-activated in transient cotransfections by a function encoded in the E2 ORF of BPV-1, albeit to a much lesser extent. In addition to activation of early viral gene transcription, the E2 regulatory gene(s) may therefore have the potential to alter cellular gene expression. PMID- 3034573 TI - Regulation of SV40 DNA replication by phosphorylation of T antigen. AB - The role of phosphorylation in regulating the biochemical properties of SV40 large T antigen has been examined. Treatment of purified T antigen with calf intestinal alkaline phosphatase resulted in the removal of 80% of the 32P label. This partially dephosphorylated T antigen displayed an increase in its ability to support DNA replication in vitro. This increase in replication activity was paralleled by an activation of specific DNA binding to site II, a necessary element within the origin of SV40 DNA replication. In contrast, the ATPase activity of dephosphorylated T antigen remained unchanged. These results demonstrate that DNA replication is regulated by phosphorylation of an origin specific DNA binding protein. PMID- 3034574 TI - Contactpoint analysis of the HeLa nuclear factor I recognition site reveals symmetrical binding at one side of the DNA helix. AB - Nuclear factor I (NFI) is a HeLa sequence-specific DNA-binding protein that is required for initiation of adenovirus (Ad) DNA replication and may be involved in the expression of several cellular genes. The interaction between NFI and its binding site on the Ad2 origin has been studied. Methylation interference and protection, u.v. irradiation of 5-BrdU substituted DNA and ethylation interference revealed major groove contacts with G and T, and phosphate backbone contacts. Computer stereographics show that the contacts are located in two blocks showing dyad symmetry to each other and 22 out of 23 contacts are accessible from one side of the helix. Inversion of the NFI binding site did not change the NFI dependent stimulation of Ad2 DNA replication in a reconstituted system. All data are compatible with NFI binding as a dimer at one side of the DNA helix. PMID- 3034576 TI - A novel leader peptide which allows efficient secretion of a fragment of human interleukin 1 beta in Saccharomyces cerevisiae. AB - Killer strains of Kluyveromyces lactis secrete a toxin which presumably is processed during secretion from a larger precursor. Analysis of the sequence of the K. lactis killer toxin gene predicts that the first 16 amino acids at the amino terminus of the protein should represent its leader peptide. We have tested the capability of this leader peptide to direct secretion of a protein fused to it by inserting a synthetic oligonucleotide identical to the sequence of the putative leader peptide into a yeast expression vector. Subsequently, the cDNA coding for the secreted active portion of the human interleukin 1 beta (IL-1 beta) was fused to the leader peptide sequence of the killer toxin. This construction in Saccharomyces cerevisiae is capable of directing synthesis and secretion of correctly processed IL-1 beta into the culture medium. PMID- 3034575 TI - Related functional domains in virus DNA polymerases. AB - Analysis of the lesions in several drug-resistant DNA polymerase mutants of herpes simplex virus along with comparative analysis of the published polymerase sequences of other human herpesviruses has shown that most lesions (five out of six) are substitutions at amino acid residues conserved in all four polymerases. Furthermore, the majority of lesions are in regions of the polypeptide where there are marked clusterings of conserved residues. On the basis of these data we have identified several domains within the polypeptide which we believe may have important functional roles in the action of the enzyme. The apparent restriction in the potential sites of lesions conferring drug resistance may explain the difficulty in selecting such mutants using acyclovir (ACV) in culture and their failure to emerge so far during ACV therapy. Extension of the comparative analysis to the polymerases of adenovirus type 2, vaccinia virus and phage phi 29 suggests that these enzymes also possess domains homologous to those most conserved in the herpes polymerases (regions I-III) and that these domains have a similar linear spatial distribution on the polypeptides. The results are discussed in relation to the known function of the DNA polymerases. PMID- 3034578 TI - Titration of DnaA protein by oriC DnaA-boxes increases dnaA gene expression in Escherichia coli. AB - Binding of the DnaA protein to its binding sites, the DnaA-boxes (TTATCCACA), was measured by a simple physiological approach. The presence of extra DnaA-boxes in growing cells leads to a derepression of dnaA gene expression, measured as beta galactosidase activity of a dnaA-lacZ fusion polypeptide. Different DnaA-boxes caused different degrees of derepression indicating that the DnaA protein requires sequences in addition to the DnaA-box for efficient binding. The DnaA boxes in oriC might act cooperatively in binding of the DnaA protein. The derepressed levels of DnaA protein obtained in a strain carrying an oriC+-pBR322 chimera were very high and sufficient to activate oriC on the chimeric plasmid, which was maintained at a copy number more than three times that of pBR322. PMID- 3034577 TI - Identification and sequence of the gene encoding cytochrome c heme lyase in the yeast Saccharomyces cerevisiae. AB - Mitochondrial cytochrome c contains a heme group covalently attached through thioether linkages to two cysteinyl residues of the protein. We demonstrate here that the nuclear gene, CYC3, in the yeast Saccharomyces cerevisiae, encodes cytochrome c heme lyase (CCHL), the enzyme catalyzing the attachment of heme to apocytochrome c. Mitochondrial extracts from cyc3- mutants are deficient in CCHL activity compared with extracts from normal strains, whereas strains carrying multiple copies of the CYC3 gene exhibit high levels of the activity. The CYC3 gene was cloned by functional complementation of a cyc3- mutant using a previously isolated plasmid containing the gene PYK1, which is tightly linked to CYC3. An open reading frame encoding a protein of 269 amino acids was identified from the DNA sequence of a fragment encompassing the CYC3 gene, and the corresponding transcript shown to be approximately 0.9 kb in length. CCHL appears to be a single polypeptide chain which acts specifically on the two forms of cytochrome c, but not on cytochrome c1. PMID- 3034580 TI - Unique pattern of point mutations arising after gene transfer into mammalian cells. AB - We have used a simian virus 40 (SV40)-based shuttle vector, pZ189, to analyze the sequence specificity of spontaneous point mutations that arise after transfection of this vector into monkey cells. The majority of the mutants which we studied had multiple base substitutions (mostly G-C----A-T transitions and G-C----T-A transversions) within the 160-bp region sequenced. Almost all of the mutations occurred in the right-hand G-C bp of one of the two following sequences, 5'-TC 3':3'-AG-5' or 5'-CC-3':3'-GG-5'. We postulate that these mutations result from DNA replication infidelity occurring during repair of the transfected DNA which has been damaged by cellular nucleases. The sequence specificity of the mutations suggests an effect of the following nucleotide on misincorporation wherein A (or less frequently T) is preferentially misincorporated opposite C when the next nucleotide inserted is A (or less frequently G). Our results support the utility of the shuttle vector as a model in studies on gene transfer and document the extreme plasticity of DNA transfected into mammalian cells. PMID- 3034579 TI - Effect of heat shock on protein degradation in mammalian cells: involvement of the ubiquitin system. AB - Exposure of cultured rat hepatoma (HTC) cells to a 43 degrees C heat shock transiently accelerates the degradation of the long-lived fraction of cellular proteins. The rapid phase of proteolysis which lasts approximately 2 h after temperature step-up is followed by a slower phase of proteolysis. During the first 2 h after temperature step-up there is a wave of ubiquitin conjugation to cellular proteins which is accompanied by a fall in ubiquitin and ubiquitinated histone 2A (uH2A) levels. Upon continued incubation at 43 degrees C the levels of ubiquitin conjugates fall with a corresponding increase of ubiquitin and uH2A to initial levels. The burst of protein degradation and ubiquitin conjugation after temperature step-up is not affected by the inhibition of heat shock protein synthesis. Cells of the FM3A ts85 mutant, which have a thermolabile ubiquitin activating enzyme (E1), do not accelerate protein degradation in response to a 43 degrees C heat shock, whereas wild-type FM3A mouse cells do. This observation indicates that the ubiquitin system is involved in the degradation of heat denatured proteins. Sequential temperature jump experiments show that the extent of proteolysis at temperatures up to 43 degrees C is related to the final temperature and not to the number of steps taken to attain it. Temperature step up to 45 degrees C causes the inhibition of intracellular proteolysis. We propose the following explanation of the above observations. Heat shock causes the conformational change or denaturation of a subset of proteins stable at normal temperatures.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3034581 TI - Human ribophorins I and II: the primary structure and membrane topology of two highly conserved rough endoplasmic reticulum-specific glycoproteins. AB - Ribophorins I and II represent proteins that are postulated to be involved in ribosome binding. They are abundant, highly-conserved glycoproteins located exclusively in the membranes of the rough endoplasmic reticulum. As the first step in the further characterization of the structure and function of these proteins, we have isolated and sequenced full-length human cDNA clones encoding ribophorins I and II using probes derived from a human liver expression library cloned into pEX1. The authenticity of the clones was verified by overlaps in the protein sequence of N-terminal and several internal fragments of canine pancreatic ribophorins I and II. The cDNA clones hybridize to mRNA species of 2.5 kb in length, and encode polypeptides of 68.5 and 69.3 kd, respectively. Primary sequence analysis, coupled with biochemical studies on the topology, indicates that both ribophorins are largely luminally disposed, spanning the membrane once and having 150 and 70 amino acid long cytoplasmically disposed C termini, respectively. Both are synthesized as precursors having cleavable signal sequences of 23 (ribophorin I) and 22 (ribophorin II) amino acids. The topology suggested by the primary structure has been confirmed biochemically using proteolytic enzymes and anti-ribophorin antibodies. Proteolysis of intact microsomes with a variety of enzymes resulted in a reduction in the apparent mol. wt of ribophorin I that would correspond to a loss of its 150-amino acid cytoplasmic tail. In the case of ribophorin II, it is completely resistant to such proteolysis which is consistent with its luminal disposition and fairly hydrophobic C terminus.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3034582 TI - The molecular organization of the H-2K region of two t-haplotypes: implications for the evolution of genetic diversity. AB - The genetic diversity between the t12 and tw5 haplotype chromosomes was studied by analyzing the molecular organization of the H-2K region. Twenty-one cosmid clones spanning over 150 kb of the H-2K region of both t-haplotypes were defined, and high resolution restriction maps were determined. Detailed comparison of the t12 and tw5 restriction maps revealed the following. (i) The H-2K regions of both t-haplotypes retain a very similar molecular organization to that reported for B10, BALB/c and AKR. The nucleotide sequence diversity estimated from restriction site polymorphism is 0.68% between the t12 and tw5 haplotypes; these two t haplotypes are no more similar to one another than BALB/c is to AKR. (ii) Genetic recombination is strongly implicated in generating H-2 polymorphism. (iii) Genetic polymorphisms, defined as small restriction fragment size differences, are observed at multiple sites along the H-2K region. An Alu-like B2 sequence and BAM5-R homologous sequence were identified as the inserted/deleted DNA segments of two of these sites, suggesting that insertion/deletion of mobile elements is a general mechanism for generating genetic diversity. PMID- 3034583 TI - Inactivation of the maize transposable element Activator (Ac) is associated with its DNA modification. AB - The Activator (Ac) element at the waxy locus (wx-m7 allele) has the ability to undergo changes in its genetic activity and cycles between an active and inactive phase. Comparison of active Ac elements at several loci and the inactive Ac at wx m7 by Southern blot analysis revealed that the inactive Ac sequence was not susceptible to digestion by the methylation sensitive enzyme PvuII while active elements were susceptible to PvuII digestion. Restriction digest comparisons between the clones of the active and inactive Ac elements were indistinguishable. Further analyses with the enzymes SstII and the methylation sensitive and insensitive isoschizomers EcoRII and BstNI showed the inactive Ac sequence was methylated at these sites, whereas the active Ac was hypomethylated. Although the active Ac at the wx-m7 allele in different genetic backgrounds showed differences in the Ac DNA modification pattern, at least a fraction of genomic DNA contained Ac sequences that were unmethylated at all of the internal sites we assayed. These data may suggest a role for DNA modification in the ability of Ac to transpose from the waxy locus and to destabilize unlinked Ds elements. PMID- 3034584 TI - Insulin and rabbit anti-insulin receptor antibodies stimulate additively the intrinsic receptor kinase activity. AB - This paper describes the properties of rabbit polyclonal antibodies directed against purified human insulin receptor which strongly stimulate the intrinsic tyrosine kinase activity. The stimulatory effect of the antibodies on the kinase activity was obtained on the insulin receptor autophosphorylation as well as on the kinase activity towards a synthetic substrate. This stimulation is additive to that induced by insulin. Moreover, rabbit antibodies do not impair insulin binding. These data strongly suggest that antibodies and insulin act through separate pathways. This conclusion is reinforced by the differences observed on the phosphopeptide maps of the receptor's beta subunit whose phosphorylation was performed either in the presence of insulin or rabbit antibodies. Interestingly, these polyclonal antibodies can also induce an activation of the receptor autophosphorylation by interacting only with extracellular determinants. The anti insulin receptor antibodies mimic insulin in their stimulatory effect on amino acid (AIB) uptake, but they have a different effect to that found on the kinase activity; the simultaneous addition of the antiserum and insulin failed to stimulate this amino acid transport over the level induced by a saturating concentration of hormone. PMID- 3034585 TI - Expression and rescuing of a cloned human tumour necrosis factor gene using an EBV-based shuttle cosmid vector. AB - A cosmid vector carrying the Epstein-Barr virus origin of replication, the EBNA-1 gene, the hygromycin phosphotransferase (hph) gene and pBR322 sequences has been constructed. This cosmid can replicate autonomously in the nucleus of human tissue culture cells, even when it carries a 35-kb long insert. The cosmid can be rescued from the transfected cells by cloning it directly into ampicillin sensitive Escherichia coli. A gene for human tumour necrosis factor (TNF) cloned into this cosmid vector was introduced in tissue culture cells, where it was transcribed into mature mRNA. PMID- 3034586 TI - The hormone response element of the mouse mammary tumour virus DNA mediates the progestin and androgen induction of transcription in the proviral long terminal repeat region. AB - Mouse mammary tumour virus (MMTV) gene expression has been shown to be regulated by glucocorticoids. A hormone response element (HRE) located between -202 and -59 upstream of the start of transcription in the long terminal repeat (LTR) region of the proviral DNA is required for this induction. We have investigated the role played by the HRE in the induction of MMTV LTR transcription by other classes of steroid hormones. Chimaeric constructs containing the HRE and the authentic LTR promoter linked to an indicator gene or the HRE linked to an otherwise hormone insensitive promoter directing the transcription of an indicator gene, were transfected into the human mammary tumour cell line T47D. Transcription at the MMTV LTR promoter or at the previously hormone-insensitive promoter was induced by progestins and androgens but not by oestradiol in transfected cells that contained functional receptors for these hormones. These results identify the HRE as the cis-acting element that mediates the progestin and androgen induction of MMTV LTR transcription. The HRE is therefore a DNA element that is required not just for glucocorticoid but also for progesterone and androgen induction of MMTV LTR transcription. PMID- 3034587 TI - Orientation and molecular map position of the complement genes in the mouse MHC. AB - Over the past few years six gene clusters have been isolated from the major histocompatibility complex (MHC) of the BALB/c mouse encompassing a total of 1600 kb of DNA and 48 genes. The molecular distances between these gene clusters and the orientation of four of the six clusters on chromosome 17 is not known. Here we use pulse-field gradient gels and Southern blot hybridization to establish large-scale genomic restriction maps covering several hundreds of kb surrounding the three gene clusters located in the K, I, S, and D regions of the MHC. Comparison of the maps orients the complement gene clusters in the S region with the 21-OHB gene pointing towards the K end and the C2 gene pointing towards the D end of the MHC. The distances between the E alpha and 21-OHB genes is 430 kb and between the C2 and TNF-alpha genes at least 420 kb. PMID- 3034588 TI - Expression of retroviral vectors in transgenic mice obtained by embryo infection. AB - Pre-implantation embryos were infected with the retroviral vector MMCV-neo, which carries the neomycin resistance (neo) gene and the v-myc gene. Three transgenic substrains (M-TKneo 1-3) were derived which stably transmit a single intact copy of the vector. In all of the substrains, expression of the neo gene from the internal thymidine kinase (TK) promoter was detected, with two of the substrains expressing the gene in all tissues analysed. In the third substrain, the vector had integrated on the X chromosome and neo expression varied between different tissues. A second series of transgenic mice were obtained with the retroviral vector SAX, in which the human adenosine deaminase cDNA (ADA) is under the control of an internal SV40 promoter. Four substrains (M-SAX 1-4) were analysed; however, no expression of the ADA cDNA was detected. In all mice, no expression was found of the genes under the control of the viral 5' long terminal repeats (LTRs). In the M-TKneo substrains the vector was hypomethylated irrespective of its expression whereas in the M-SAX mice the vector was hypermethylated. These results demonstrate for the first time that the TK promoter can apparently express a gene in all tissues of adult mice and that retroviral vectors with internal promoters may provide an alternative to DNA injection for the efficient expression of genes in transgenic mice. PMID- 3034589 TI - Identification of two distinct elements in the long terminal repeat of HTLV-I responsible for maximum gene expression. AB - Human T-cell leukemia virus type I has a unique sequence, pX, between env and the 3' long terminal repeat (LTR). One of its products, p40, activates gene expression directed by the LTR in a trans-acting manner. We have analysed the mechanism of this trans-activation mediated by p40 in human T cells co transfected with a plasmid expressing p40 using the transient CAT gene expression. We identified two distinct elements in the LTR which are involved in maximum gene expression. The first was present in a 230-bp fragment upstream from TATA box in the U3 region and behaved as a classical enhancer. This region was also shown to be responsible for trans-activation by p40. This element alone together with functional p40 could direct the gene expression at only approximately 10% of the level achieved by the complete LTR and p40. The second element was present within a 300-bp fragment downstream from the RNA start site and profoundly enhanced the gene expression in a way independent from trans activation mechanism. This enhancement was observed only when the element was located immediately downstream from the RNA start site without orientation preference. These two elements participate independently in the enhancement of gene expression. PMID- 3034590 TI - Relationship between enhanced reactivation and mutagenesis of u.v.-irradiated human cytomegalovirus in normal human cells. AB - The survival of u.v.-irradiated human cytomegalovirus (HCMV) on u.v.-irradiated human IAFP-1 cells was increased over that on unirradiated cells. Irradiated virus had a higher forward mutation frequency towards temperature sensitivity in irradiated than in unirradiated cells. Enhanced reactivation of u.v.-irradiated HCMV is thus mutagenic in normal human cells. This observation supports the possible induction of an error-prone mode of DNA repair in u.v.-irradiated mammalian cells. PMID- 3034591 TI - Developmental expression of Drosophila melanogaster retrovirus-like transposable elements. AB - We have determined the pattern of temporal expression of several Drosophila retrovirus-like transposable elements. Some of these elements can be grouped into classes whose members show a similar profile of developmental transcription. The members of the 412 class, which includes 412, mdg1, 17.6 and 3S18, are transcribed mainly in the early larval and pupal stages of development, with small differences among the various members. HMS Beagle and Springer constitute another class where RNA accumulation in the larval stages is higher than in pupae and the adult flies accumulate more RNA than any other stage of development. Finally, the transcription of other elements such as copia, 297 and B104 follows a specific and individual pattern distinct from those described above. These results suggest the existence of evolutionary relationships among different transposable elements in Drosophila and the involvement of different cellular genes in the control of their expression. PMID- 3034592 TI - Recombinant forms of M13 procoat with an OmpA leader sequence or a large carboxy terminal extension retain their independence of secY function. AB - The assembly of phage M13 procoat protein into the plasma membrane of Escherichia coli is independent of the secY protein. To test whether this is caused by the unusually small size of procoat, we fused DNA encoding 103 amino acids to the carboxy-terminal end of the procoat gene. The resulting fusion protein, which attains the same membrane-spanning conformation as mature coat protein, still does not require the secY function for membrane assembly. To determine whether the leader sequence governs interaction with the secY protein, we genetically exchanged the leader peptides between procoat and pro-OmpA, a protein which does require secY for its membrane assembly. Each of the resulting hybrid proteins assembles across the plasma membrane, though at a reduced rate. Membrane assembly of the fusion of procoat leader and OmpA required secY function, whereas assembly of the pro-OmpA leader/coat protein fusion was independent of secY. Properties of the entire procoat molecule, rather than its small size or a specific property of its leader peptide determines its mode of membrane assembly. PMID- 3034593 TI - Unusual properties of promoter-up mutations in the Escherichia coli galactose operon and evidence suggesting RNA polymerase-induced DNA bending. AB - Two mutations are described, each of which renders the Pribnow box sequence of one of the two overlapping promoters of the Escherichia coli galactose operon identical to the consensus sequence TATAAT. Both double exchanges were specifically introduced into the original context by oligonucleotide-directed mutation construction. Each of the mutant promoters exhibits a greatly enhanced capacity to form stable complexes with RNA polymerase, as judged by nuclease protection experiments and by assaying shifts of electrophoretic mobility. On the other hand, the effect of the same mutations on the rates of transcription from the two gal promoters is strikingly different. Unexpectedly, when complexed with RNA polymerase, DNA fragments carrying one of the two double exchanges were found to differ from each other as well as from the corresponding wild-type fragment with respect to their electrophoretic mobilities. These observations are indicative of different three-dimensional structures of these complexes which may reflect different forms of DNA bending induced in these otherwise identical fragments by complex formation with RNA polymerase. PMID- 3034594 TI - The GABAA/benzodiazepine receptor is a heterotetramer of homologous alpha and beta subunits. AB - The GABAA receptor has been purified to homogeneity from bovine cerebral cortex. Under stringent conditions of isolation, the GABAA receptor was shown to consist only of alpha (Mr 53 000) and beta (Mr 57 000) subunits. A densitometric scan of SDS-PAGE gels under reducing conditions showed that these subunits were present in a 1:1 ratio. A model of the receptor as a heterologous tetramer alpha 2 beta 2 is proposed. Monoclonal antibodies have been raised to the purified bovine GABAA receptor. One of these antibodies, 1A6, was shown to react with both the alpha and beta subunits of the purified receptor. The subunits were still positive in immunoblots following the removal of the carbohydrate moieties of the respective polypeptides by endoglycosidase F treatment. This antibody has been employed to demonstrate antigenic cross-reactivity between the GABAA receptors of three vertebrate species. It is further proposed that there is partial amino acid sequence homology between the alpha and beta polypeptides and hence that they are derived from a single ancestral gene. PMID- 3034596 TI - Single amino acid changes that render human IFN-alpha 2 biologically active on mouse cells. AB - Human IFN-alpha 1 and IFN-alpha 2 differ in 28 of 166 amino acids and show very different specific antiviral activities on human and murine cells. We have identified, by hybrid scanning and site-directed mutagenesis, three residues in IFN-alpha 2, in positions 121, 125 and 132 which, when replaced individually or jointly by their IFN-alpha 1 counterparts, modify its activity on mouse cells by up to 400-fold. We argue that these residues are involved in direct contacts with the mouse interferon receptor. PMID- 3034595 TI - Immunoglobulin heavy chain switch region recombination within a retroviral vector in murine pre-B cells. AB - We have employed a retroviral vector, ZN(Smu/S gamma 2b)tk1, as a substrate for detecting the presence of immunoglobulin heavy chain constant region (CH) gene switch (S) recombination activity in murine pre-B cells. ZN(Smu/S gamma 2b)tk1 contains a neomycin (neo) resistance gene in addition to the herpes simplex virus thymidine kinase (Htk) gene which is positioned between murine Smu and S gamma 2b sequences. Stable acquisition of the ZN(Smu/S gamma 2b)tk1 vector was selected in G-418 and switch region recombination within these proviruses was selected by resistance to the drug bromodeoxyuridine (BUdR). Fluctuation analyses of ZN(Smu/S gamma 2b)tk1 infected 18-8tk- and 38B9tk- pre-B lines revealed Htk gene inactivations with apparent frequencies of 5 X 10(-5) and 1 X 10(-5) events/cell/generation, respectively, while G-418 resistant Ltk- fibroblasts lost the HTK phenotype at an apparent rate of 4 X 10(-8). Southern blot analysis demonstrated that switch recombination caused the deletion of the Htk gene in all pre-B clones examined while the loss of Htk in Ltk- clones was not mediated by S region recombination. In 21 out of 24 pre-B clones, the recombinations involved the tandemly repetitive portions of the Smu and S gamma 2b sequences. These results demonstrate that the CH gene S region segments inserted into ZN(Smu/S gamma 2b)tk1 are sufficient for B-cell-specific recombination/deletion within the S region tandem repeats. PMID- 3034597 TI - Superinduction of the human interferon-beta promoter. AB - Superinduction, that is induction with simultaneous blocking of mRNA translation, enhances the induction of interferon-beta in response to virus or double-stranded RNA in human fibroblasts. Expression of the cloned IFN-beta gene upon transfer into heterologous cells reflects the endogeneous interferon expression with respect to induction or superinduction indicating the involvement of cellular mediators in this mode of gene regulation. Expression from gene hybrids in mouse Mo57/2 cells reveals that 5' flanking DNA sequences from the human IFN-beta gene are responsible for induction and superinduction. Superinduction of the human IFN beta promoter is demonstrated in several rodent and primate cell lines. In addition, expression from promoter mutants in mouse cells indicates that DNA sequences responsible for induction and superinduction are identical. PMID- 3034598 TI - Overexpression of the EGF receptor-related proto-oncogene erbB-2 in human mammary tumor cell lines by different molecular mechanisms. AB - Amplification of the erbB/EGF receptor and a structurally related gene, designated erbB-2, have previously been detected in a variety of human tumors. In a series of human mammary tumor cell lines, analysis of transcripts of these genes revealed elevated levels of one or the other in more than 60% of tumors analyzed. Eight cell lines demonstrated erbB-2 mRNA levels ranging from 4- to 128 fold above those of normal controls. erbB-2 expression was evaluated in comparison to the expression level of actin observed in these cell lines. There was no evidence of an aberrantly sized erbB-2 transcript in any of these lines. Immunoblot analysis indicated elevation in levels of the 185-kd product of the erbB-2 gene expressed by these cells. In four lines erbB-2 gene amplification in the absence of an apparent gene rearrangement was demonstrated. In a representative cell line of this type, SK-BR-3, the amplified erbB-2 gene copies were located in an aberrant chromosomal location. Four additional cell lines, which demonstrated 4- to 8-fold overexpression of erbB-2 mRNA, did not exhibit gene amplification. In a representative cell line of this type ZR-75-1, an apparently normal chromosomal location was found for the erbB-2 gene. Our findings indicate that overexpression of the erbB-2 gene in mammary tumor cell lines is frequent and associated with different genetic abnormalities. PMID- 3034599 TI - Characterization of recombinant human granulocyte-colony-stimulating factor produced in mouse cells. AB - Mouse C127I cells were transformed with a chimeric plasmid consisting of bovine papillomavirus DNA and human granulocyte-colony-stimulating factor (G-CSF) cDNA placed under the control of the SV40 early promoter. The transformed cells secreted constitutively a high level of human G-CSF, 10-20 micrograms/ml in a low serum medium. The secreted G-CSF has been purified to homogeneity by a two-step procedure including gel filtration and hydrophobic column chromatography. The purified recombinant G-CSF runs as a single band with an apparent Mr of 19,000 on a polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. This value corresponds to that of the native human G-CSF purified from the medium conditioned by human carcinoma CHU-2 cells. The recombinant human G-CSF was as active as native G-CSF in vitro in supporting proliferation of mouse NFS-60 cells and stimulating colony formation from human as well as mouse bone marrow cells. When the recombinant human G-CSF was subcutaneously administrated into mice, a remarkable stimulation of granulopoiesis and splenomegaly was observed. PMID- 3034600 TI - Close genetic and physical linkage between the murine haemopoietic growth factor genes GM-CSF and Multi-CSF (IL3). AB - The two murine haemopoietic growth factors, granulocyte-macrophage colony stimulating factor (GM-CSF) and Multi-CSF (interleukin 3) stimulate the proliferation and differentiation of an overlapping set of haemopoietic progenitor cells and are produced coordinately following activation of T lymphocytes. Here we report the chromosomal location of the genes encoding these two factors. Initially both genes were assigned to chromosome 11 by analysis of mouse/Chinese hamster somatic cell hybrids. Genetic analysis using an interspecies (Mus musculus X Mus spretus) back-cross confirmed this assignment by demonstrating that both the GM-CSF and Multi-CSF genes are genetically linked to the SPARC gene, which had been independently assigned to sub-band B1 of chromosome 11. Analysis of physical distances by pulsed field gel electrophoresis demonstrated further that the two CSF genes lie within 230 kb of each other. However examination of the subchromosomal region containing all three loci by pulsed field gel analysis showed that SPARC is at least 400-500 kb distant from the region containing the two CSF genes. PMID- 3034601 TI - Negative control of liver-specific gene expression: cloned human retinol-binding protein gene is repressed in HeLa cells. AB - It has been found that 334 bases of the 5' flanking region of the human retinol binding protein (RBP) gene contain sufficient information to direct accurate and specific transcription in human hepatoma but not in HeLa cells. Dissection of this region reveals the existence of at least three distinct controlling elements: an enhancer, which can activate promoters in a variety of cell lines and is therefore non-tissue specific; a negative cis-acting element, which apparently binds a repressor molecule not present or non-functional in hepatoma cell lines; a promoter element. Transcription is confined to hepatoma cells only when both the negative element and the promoter element are present. PMID- 3034602 TI - Molecular cloning of the beta-subunit of human prolyl 4-hydroxylase. This subunit and protein disulphide isomerase are products of the same gene. AB - Prolyl 4-hydroxylase (EC 1.14.11.2), an alpha 2 beta 2 tetramer, catalyses the formation of 4-hydroxyproline in collagens by the hydroxylation of proline residues in peptide linkages. We report here the isolation of cDNA clones coding for the beta-subunit of prolyl 4-hydroxylase from a human hepatoma lambda gt11 library and a corresponding human placenta library. Five overlapping clones covering all the coding sequences and almost all the non-coding sequences were characterized. The size of the mRNA hybridizing with these clones in Northern blotting is approximately 2.5 kb. The clones encode a polypeptide of 508 amino acid residues, including a signal peptide of 17 amino acids. These human sequences were found to be very similar to those recently reported for rat protein disulphide isomerase (EC 5.3.4.1). The degree of homology between these two proteins was 84% at the level of nucleotide sequences or 94% at the level of amino acid sequences. Southern blot analyses of human genomic DNA with a cDNA probe for the beta-subunit indicated the presence of only one gene containing these sequences. The product of a single gene thus appears to possess two different enzymatic functions depending on whether it is present in cells in monomer form or in the prolyl 4-hydroxylase tetramer. PMID- 3034603 TI - Characterization and expression of a murine gene homologous to human EPA/TIMP: a virus-induced gene in the mouse. AB - A genomic clone encompassing the entire coding region of a murine gene homologous to human erythroid potentiating activity/tissue inhibitor of metalloproteinase (EPA/TIMP) was isolated and sequenced. Based on alignment with human EPA/TIMP cDNAs we deduce a structure comprising five exons and four introns extending over 4.3 kb of DNA. In mouse and hamster cell lines transcription from this gene and interferon genes is induced by Newcastle Disease virus (NDV). Examination of the 5'-flanking sequences of the gene reveals a set of repeated elements with structural similarity to those previously described as inducer-responsive elements in the human IFN-beta 1 gene. The 4.3-kb DNA fragment encompassing the homologous murine EPA/TIMP gene was transfected into human T98G cells and transfectants tested for NDV inducibility. In contrast to the endogenous human gene, the integrated murine EPA/TIMP gene was NDV-inducible and TIMP activity was detectable in the cell culture fluid. PMID- 3034604 TI - Enzymatic activation of Fujinami sarcoma virus gag-fps transforming proteins by autophosphorylation at tyrosine. AB - Site-directed mutagenesis of the Fujinami sarcoma virus (FSV) genome has suggested that Tyr 1073 of the P130gag--fps protein-tyrosine kinase is a regulatory site. To investigate directly the ability of tyrosine phosphorylation to affect P130gag--fps kinase activity, the phosphotyrosyl phosphatase inhibitor orthovanadate and partially purified phosphotyrosyl phosphatases were used to manipulate the stoichiometry of P130gag--fps phosphorylation. Phosphorylation of P130gag--fps at Tyr 1073 correlated with enhanced kinase activity. The thermolabile phosphorylation, kinase activity and transforming ability of P140gag -fps encoded by a temperature-sensitive (ts)FSV variant were restored at the non permissive temperature for transformation by incubation of infected cells with orthovanadate. In this case tyrosine phosphorylation can apparently functionally reactivate a conditionally defective v-fps kinase activity. These data suggest that reversible autophosphorylation at a conserved tyrosine within the v-fps kinase domain is a positive regulator of enzymatic activity and biological function. Phenotypic suppression of the tsFSV genetic defect by orthovanadate emphasizes the potential importance of phosphotyrosyl phosphatases in antagonizing tyrosine kinase action. It is suggested that autophosphorylation may constitute a molecular switch by which some protein-tyrosine kinases are activated. PMID- 3034605 TI - Production of hepatitis B virus in vitro by transient expression of cloned HBV DNA in a hepatoma cell line. AB - Transfection of human hepatoma cell lines with cloned HBV DNA resulted in the secretion of large amounts of hepatitis B surface antigen (HBsAg) and core related antigens (HBc/HBeAg) if well-differentiated cell lines were employed. Synthesis of both viral antigens was the highest in cell line HuH-7 and continued for approximately 25 days. Particles resembling hepatitis B virions (Dane particles) by morphology, density and by the presence of the preS1 surface antigen were released from the transfected HuH-7 cells into the culture medium. These particles produced in vitro were also indistinguishable from the naturally occurring hepatitis B virions in containing the virus-associated DNA polymerase and mature HBV genomes. Restriction analysis of these DNA molecules was compatible with the nucleotide sequence of the transfecting HBV DNA sequence. Viral surface antigens and core proteins present in the culture medium were fractionated and characterized by immunoprecipitation and SDS--PAGE after labeling with [35S]methionine. Antisera specific for X-gene products identified in cell extracts two hitherto unknown HBV gene products. This system thus provides a new approach to open questions regarding HBV-related gene function and HBV replication. PMID- 3034606 TI - Import of frog prepropeptide GLa into microsomes requires ATP but does not involve docking protein or ribosomes. AB - Frog prepropeptide GLa, a precursor to a secretory protein containing 64 amino acids, was processed and imported by dog pancreas microsomes. These events did not depend on either docking protein or on the presence of ribosomes. A hybrid protein between the first 60 amino acids of prepropeptide GLa and an unrelated peptide of 49 amino acids fused to the carboxy terminus, however, behaved like a typical secretory protein precursor with regard to docking protein dependence. This suggests that independence of the need for docking protein, in the case of prepropeptide GLa, can be attributed to the size of the precursor protein. Processing and import of prepropeptide GLa by microsomes were ATP dependent. Therefore, import of proteins into the endoplasmic reticulum (ER) includes an ATP requiring step not involving a ribosome/ribosome receptor or signal recognition particle (SRP)/docking protein interaction. PMID- 3034607 TI - Three suppressor mutations which cure a mitochondrial RNA maturase deficiency occur at the same codon in the open reading frame of the nuclear NAM2 gene. AB - Dominant mutations of the nuclear NAM2 gene are able to compensate for a deficiency of the maturase encoded by the fourth intron of the mitochondrial cytochrome b gene. We have determined the complete nucleotide sequence of the NAM2-1 suppressor allele. The results of S1 nuclease protection experiments show that two overlapping poly(A)+ RNAs are transcribed from the gene using different promoters. The longer transcript contains two open reading frames (ORFs), a long ORF which could encode a protein of 894 amino acids, mol. wt 102,000 daltons, and a short ORF of 51 codons which is omitted from the shorter transcript. The wild type nam2+ and two other suppressor alleles, NAM2-6 and NAM2-7, have been cloned. A comparison of the sequence of the wild-type and the three suppressor alleles shows that on three separate occasions the same codon specifying glycine was mutated (once to serine and twice to cysteine). Finally sequence comparisons identified two regions in the long ORF, distinct from the position of the suppressor mutations, that could correspond to binding domains for a nucleotide and a nucleic acid. PMID- 3034609 TI - Yeast DNA polymerase--DNA primase complex; cloning of PRI 1, a single essential gene related to DNA primase activity. AB - The immunopurified yeast DNA polymerase--DNA primase complex is constituted by DNA polymerase I polypeptides and by three other protein species, called p74, p58 and p48, which we show to be immunologically unrelated. The gene encoding the p48 polypeptide has been identified by immunological screening of a lambda gt11 yeast genomic DNA library. Antiserum specific for p48 inhibits DNA primase, and immunoreactive, inhibitory antibodies are affinity-purified by the clone-encoded protein, thus relating the p48 polypeptide to DNA primase activity. The entire gene has been cloned, and the 1.45-kb p48 mRNA is overproduced in cells containing the gene in high copy number. Gene disruption and Southern hybridization experiments demonstrate that the p48 protein is encoded by a single gene and it performs an essential function. PMID- 3034608 TI - The product of the mei3+ gene, expressed under control of the mating-type locus, induces meiosis and sporulation in fission yeast. AB - In fission yeast the ability to undergo meiosis and sporulation is conferred by the matP+ and matM+ genes of the mating-type locus. Inactivation of ran1+, a negative regulator of meiosis, is thought to be an essential step in meiotic initiation. We have isolated a further meiotic control gene mei3+, and have shown the following: a null allele of mei3 totally inhibits meiosis; the mei3+ RNA transcript and its translational product are expressed only in matP+/matM+ diploids entering meiosis; forced expression of mei3+ in vegetative cells provokes haploid meiosis and sporulation. We suggest that the product of mei3+ gene, a protein of 21 kd, initiates meiosis by inactivating ran1+. PMID- 3034611 TI - Contacts between gamma delta resolvase and the gamma delta res site. AB - We have investigated the interaction between resolvase and the res site of the transposon gamma delta by methylation and ethylation interference experiments. We have examined the effect of these DNA modifications both on binding and resolution in vitro. Major groove methylations within a 9 bp sequence that borders each site inhibit binding of resolvase to that site. Ethylation of certain phosphates within, and adjacent to, this border sequence inhibits binding. Together, these interference points define a contact region, present at all three res sites. In vitro resolution is inhibited only by modifications within site I. Inhibition of resolution by methylation of adenines at the center of site I suggests that minor groove contacts near the crossover may be required for resolution activity. PMID- 3034610 TI - Autoregulation of bacteriophage P2 repressor. AB - The immunity repressor of bacteriophage P2 regulates the two convergent promoters, Pe and Pc, located in the early control region. Pe is the early promoter which is negatively regulated by the repressor. It was found, by DNase I protection studies, that the P2 repressor enhances the binding of RNA polymerase to Pc. Furthermore, under in vivo conditions the transcription initiated at Pc, measured as chloramphenicol acetyl transferase gene expression, is low in the absence of repressor but is stimulated by low repressor levels. With increasing repressor concentrations transcription from the Pc promoter decreases. Thus, the P2 repressor both negatively and positively regulates its own promoter. PMID- 3034612 TI - Effect of atracurium on twitch, tetanic and post-tetanic twitch responses at the rat neuromuscular junction. AB - The effect of atracurium on the phenomenon of post-tetanic potentiation, which is believed to be of a presynaptic origin, i.e. due to endogenous transmitter release, was investigated to see if atracurium had a presynaptic inhibitory mechanism at the rat neuromuscular junction. The results showed that atracurium (0.8-80 microM) reduced the indirectly elicited twitch, tetanic and post-tetanic twitch tensions, in a dose-dependent manner. Atracurium (1 microM) produced a tetanic fade in the preparations stimulated at 20 Hz and above. The acetylcholine (ACh) released at high frequencies of nerve stimulation was collected, in the presence of physostigmine (0.77 microM), added to a rat ileum preparation, in which it produced a small contraction which was blocked by atracurium (1 microM), which in turn blocked the contracture produced by ACh (1 microM) added directly to the organ bath. It was concluded that, in addition to its well-known competitive blockade of postsynaptic ACh receptors, atracurium may also have a presynaptic inhibitory effect at the neuromuscular junction. PMID- 3034613 TI - Combined evaluation of first pass radionuclide angiography and equilibrium radionuclide ventriculography in the diagnosis of coronary artery disease. II. Results during exercise. AB - Results of 203 patients who underwent first pass radionuclide angiography (FP) and quantitative equilibrium radionuclide ventriculography (qERNV) were stored in a data base system and evaluated statistically. Eighty eight of these patients also underwent exercise equilibrium radionuclide ventriculography (E-qERNV). In patients with coronary artery disease (CAD) without previous myocardial infarction (MI), evaluation of global and regional ejection fraction (gEF, rEF) at rest revealed a poor sensitivity of 64%, the specificity was about 71% (qERNV). FP at rest revealed similar values of sensitivity (69%) and specificity (83%). Additional assessment of stress induced changes of gEF, significantly (P less than 0.05) improved sensitivity of qERNV in CAD patients without a history of previous MI to 84% (specificity 86%). In patients with one previous MI, however, similar values of sensitivity were found (R-FP: 87%, R-qERNV: 84%, E qERNV: 93%). In patients with several MI's, sensitivity was above 90% at rest and during exercise (R-FP: 96%, R-qERNV: 93%, E-qERNV: 100%). PMID- 3034614 TI - Preparation and evaluation of 99mTc(V)-DMSA complex: studies in medullary carcinoma of thyroid. AB - Consequent to the promising results reported with 99mTc(V)-DMSA for imaging certain types of soft tissue tumors, we have developed methods to prepare this radiopharmaceutical in three ways: from freshly prepared reagents, through the use of a two component kit and use of the standard renal DMSA kit by a modified recipe. The 99mTc(V)-DMSA complex has been subjected to paper electrophoretic and chromatographic procedures and also biodistribution studies. The distinctly different behaviour of this new product compared to that of the well known renal DMSA complex has been clearly established. Scintiimaging in a preliminary clinical trial in patients with medullary carcinoma of the thyroid has been encouraging. PMID- 3034615 TI - Renal washout and vascular resistances. AB - Evaluation of the vascular transit through a transplanted kidney depends on the convolution of the arterial bolus in the renal vasculature. In order to stress the importance of the renal artery, an example where a moderate stenosis resembled allograft rejection is given. Interpretation of flow studies could reveal a stenosis if allograft biopsy is performed at an early stage. PMID- 3034616 TI - The importance of primary cytomegalovirus infection in childhood cancer. AB - Sixty-eight paediatric patients with malignant tumours or leukaemia were followed for signs of infection with human cytomegalovirus (HCMV) over 1 year. HCMV was isolated from 24 out of 68 patients at some point during the observation period; from urine in 14, from both urine and throat in 9 patients, and from throat alone in 1 patient. Previous antibody analysis indicated the presence of HCMV antibodies in 10 of the 24 virus-shedding patients, while 7 patients were seronegative and 7 undefined. Thus the incidence of reactivation appears to be higher than that of primary infection in these immunocompromised patients. The mean duration of virus shedding was 4.2 months in the primary infection group, 1.7 months in the reactivation group and 1.1 months in the undefined group. No difference in the incidence of HCMV-associated illness was observed between patients with leukaemia and those with malignant tumours. Clinical symptoms associated with HCMV infection (pneumonia (2), fever (6) and hepatitis (1)) were observed in all patients with primary infections and in only five patients with reactivated infection. PMID- 3034618 TI - An unusual, possibly "new" MA/MR syndrome with sagittal craniosynostosis. AB - This report is on a mentally retarded, male child with multiple anomalies: sagittal craniostenosis, bilateral coloboma of the iris, craniofacial dysmorphy, asymmetrical split hand malformation, bilateral syndactyly of 2nd-4th toes and perineoscrotal hypospadias. PMID- 3034617 TI - Gastroenteritis in infants, associated with a genome type of adenovirus 31 and with combined rotavirus and adenovirus 31 infection. AB - In an infants' ward, gastroenteritis occurred in five children in two groups, probably by nosocomial spread of adenovirus 31 (three cases) and adenovirus 31 + rotavirus (two cases). The infants recovered well. The DNA of adenovirus 31 isolates was analysed with ten restriction endonucleases and found identical for all five strains, but different from the prototype. PMID- 3034619 TI - Possibility of elevated parathyroid function in patients with calcium-containing nephrolithiasis as compared with normal controls. AB - 109 patients with calcium-containing nephrolithiasis and 10 normal controls underwent oral calcium load test. After thorough examination, 6 of the 109 patients were diagnosed as having primary hyperparathyroidism (PHPT) and the remainder as having normocalcemic nephrolithiasis without PHPT. Following the oral calcium load test, the latter were operationally divided into 3 groups - normocalciuric nephrolithiasis (NN), n = 78; absorptive hypercalciuria (AH), n = 10, and renal hypercalciuria (RH), n = 15 - according to the criteria reported by Pak et al. Before the oral calcium load test, nephrogenous adenosine 3',5' monophosphate (NcAMP), urinary adenosine 3'-5'-monophosphate (urinary cAMP), and plasma immunoreactive parathyroid hormone (iPTH) were determined to evaluate parathyroid function. This function, as assessed by mean basal NcAMP in the NN, AH and RH groups as well as the PHPT group, was significantly increased as compared with that in the normal controls. Within the NcAMP-elevated 4 groups, the mean basal NcAMP was highest in the PHPT group followed by the RH, AH and NN groups. In view of the mean basal NcAMP, disregarding the PHPT group, the NN and AH groups seemed to be intermediate types between the normal controls and the RH groups. Similar, but less distinctive results were obtained in the determination of urinary cAMP and plasma iPTH. On the other hand, when leaving the PHPT group out, the mean basal urinary calcium creatinine ratio (Ca/Cr) was highest in the RH group followed by the AH and NN groups, and lowest in normal controls, suggesting that the NN and AH groups were intermediate between normal controls and the RH group. The mean basal urinary Ca/Cr ratio in the PHPT group was moderately elevated but not remarkable. Almost similar tendencies were observed in 24-hour urinary calcium excretions on a calcium-restricted diet. A weakly positive correlation (r = 0.232, p less than 0.05) between basal NcAMP and basal urinary Ca/Cr ratio was observed in accumulated cases of the NN, AH and RH groups, whereas a negative correlation (r = -0.664, p less than 0.05) was obtained in normal controls. It is concluded that a possible abnormal calcium metabolism is suggested in stone formers without PHPT. Additionally, it is speculated that 'relative hypercalciuria' in NN and hypercalciuria in AH and RH might be accounted for in a single line of a primary renal leak of calcium. PMID- 3034620 TI - Heterogeneous distribution of beta and alpha-2 adrenoceptor binding sites in human fat cells from various fat deposits: functional consequences. AB - Investigations have been carried out to explain the heterogeneity of response to catecholamines of human fat cells from various deposits. Adipocytes from two subcutaneous sites (abdominal and femoral) were studied concomitantly in women while omental fat cells were taken from another group of patients undergoing abdominal surgery. Alpha-2 and beta sites were identified in fat-cell membranes with [3H]yohimbine and [3H]dihydroalprenolol, respectively. Lipolytic responses were tested with isoproterenol (beta agonist), clonidine (alpha-2 agonist) and epinephrine. There are clear differences in the relative number of beta and alpha 2 sites according to the origin of the fat deposit; beta sites are less numerous than alpha-2 sites in subcutaneous fat cells of both regions (alpha-2:beta sites are in a ratio of 3 +/- 0.4:2 +/- 0.4). However, in membranes of omental fat cells, beta sites are at least as numerous as alpha-2 sites (ratio 0.9 +/- 0.2). Epinephrine always has a higher affinity for alpha-2 sites than for beta sites in the subcutaneous and omental deposits. In lipolysis studies, epinephrine, in the absence of adenosine in the incubation medium, initiated an anti-lipolytic effect in femoral fat cells and promoted inhibition of lipolysis at lower concentrations in abdominal subcutaneous fat cells, the effect being reversed at higher doses; epinephrine, however, was always lipolytic in omental adipocytes. There was no striking differences in the sensitivity to isoproterenol in the various deposits. Clonidine had a higher affinity for alpha-2 sites in femoral fat cells and was equipotent in the omental and abdominal ones. Thus, the differences in the lipolytic responses to epinephrine in adipocytes from different sites are linked to a variable alpha-2 inhibiting effect (and alpha-2 site number) rather than to a modified beta driven increase in lipolysis initiated by the physiological amine. PMID- 3034621 TI - Postsynaptic alpha 1- and alpha 2-adrenoceptors in human blood vessels: interactions with exogenous and endogenous catecholamines. AB - The relative efficacy of epinephrine and norepinephrine on vascular alpha 1- and alpha 2-adrenoceptors and also the difference in vasoconstriction induced by exogenous norepinephrine as opposed to neuronally released norepinephrine were studied in the forearm of healthy volunteers. Intra-arterial cumulative dose infusions of epinephrine and norepinephrine (0.6, 1.6 and 4.0 ng kg-1 min-1) were given in the presence of saline, the selective alpha 1-antagonist doxazosin (0.1 microgram kg min-1) the selective alpha 2-antagonist yohimbine (1.0 microgram kg min-1) and the combination of both antagonists. beta-Adrenoceptor-mediated effects were prevented by a concomitant i.a. infusion of propranolol (1.0 microgram kg-1 min-1). Forearm blood flow (FBF) was measured before each infusion and at the end of each dose step. Neuronal norepinephrine was released by i.a. infusion of tyramine in three cumulative doses (0.25, 0.50 and 1.25 micrograms kg 1 min-1) and by lower body negative pressure (LBNP, -40 mmHg for 5 min). Changes in FBF were measured without and with concomitant i.a. infusions of the aforementioned doses of doxazosin and yohimbine. In the LBNP experiment the opposite arm was used as a control. Forearm blood flow was measured by plethysmography. Epinephrine and norepinephrine induced an equal and dose dependent vasoconstriction, which was significantly inhibited by doxazosin as well as yohimbine and to a greater extent by the combination of the antagonists. No differences were found between epinephrine and norepinephrine in this respect.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3034622 TI - Lisinopril in hypertensive patients with and without renal failure. AB - Lisinopril (MK521), a lysine analogue of enalaprilic acid, the bioactive metabolite of enalapril, has a longer half-life than enalaprilic acid, and is excreted unchanged in the urine. Its kinetic profile and antihypertensive and hormonal effects have been investigated in an open study in 3 groups each of 6 hypertensive patients, with normal, moderate and severe impairment of renal function. Serum drug level, blood pressure, converting enzyme activity (CEA), plasma renin activity (PRA), aldosterone concentration (PAC), and serum potassium and creatinine were measured during 1 week following a single oral dose and subsequently following 8 daily doses of 5 mg lisinopril. Accumulation of lisinopril was found in the severe renal failure group. CEA was suppressed to less than 10% of its initial value from 4 to 24 h after the initial dose in all three groups, and the suppression was more marked and lasted longer in patients with severe renal failure. An inverse correlation was found in all patients between log serum lisinopril concentration and log CEA. Lisinopril lowered blood pressure in all three groups over 24 h. PRA rose and PAC fell similarly in the groups. Serum potassium increased in the renal failure groups and creatinine remained unchanged in all groups. Thus, when lisinopril 5 mg is given daily to patients with severe renal failure it may accumulate. The high serum lisinopril concentration does not cause an excessive antihypertensive effect. In patients with severe renal failure, adjustment of the dose or the dosing frequency to the degree of renal failure is recommended to avoid administration of doses in excess of those required to achieve adequate inhibition of converting enzyme. PMID- 3034624 TI - Aberrant expression of Ia-like antigens on tumor cells of regressing but not of progressing Rous sarcomas. AB - Unlike many other tumors, experimentally induced Rous sarcomas in chickens have a definite tendency to regress spontaneously. Regression is under genetic control, but it is immunologically mediated although the target antigens that stimulate an immune response are unknown. Tumor cells in regressing sarcomas were found to express class II major histocompatibility complex (Ia-like) antigens while tumor cells from progressing sarcomas were negative. This suggests that the induction of Ia-like antigen expression has a role in the initiation or perpetuation of regression, similar to that postulated for class II antigen expression in a variety of autoimmune diseases. PMID- 3034625 TI - A new V gene expressed in lambda-2 light chains of the mouse. AB - We have partially sequenced the light chain variable regions expressed in three IgM-producing hybridomas generated from newborn mice or from manipulated animals suppressed for IgM production. In these lines a new V gene (V-lambda-X), exhibiting less than 60% homology to any known lambda or kappa V gene, is rearranged to J-lambda-2. The light chains produced by these cells contain the lambda-2 constant domain, but are not recognized by goat antisera raised against conventional mouse lambda light chains. PMID- 3034623 TI - Influence of calcium entry blockade on alpha 1- and alpha 2-adrenoceptor mediated vasoconstriction in the forearm of hypertensive patients. AB - The influence of treatment with the calcium entry blockers PY 108-068 (PY) and PN 200-110 (PN) on alpha 1- and alpha 2-adrenoceptor mediated vasoconstriction has been investigated in the forearms of hypertensive patients. Changes in forearm vascular resistance (FVR) in response to the intra-arterial infusion of drugs were determined at the end of a placebo period and after 2-4 weeks of treatment with PY or PN. The drugs used were the selective agonists methoxamine (alpha 1) and B-HT 933 (alpha 2). During placebo, basal FVR was dose-dependently increased by methoxamine and B-HT 933. Basal blood pressure was lowered during PN but not during PY. Treatment with the calcium entry blockers did not influence the effect of methoxamine, but the vasoconstriction induced by B-HT 933 was attenuated by both of the calcium entry blockers. These results confirm the findings in animal studies that calcium entry blockers preferentially inhibit the alpha 2 adrenoceptor mediated vasoconstriction induced by selective agonists. PMID- 3034626 TI - A comparison of the pharmacological actions of 4-aminopyridine and two of its derivatives in the monkey. AB - The neuromuscular, cardiovascular and central nervous system stimulating effects of 4-aminopyridine (4-AP), 2,4-diaminopyridine (2,4-DAP) and LF-14 were investigated in the monkey. All these compounds were shown to reverse the stable neuromuscular blockade produced by the intravenous infusion of pancuronium bromide. The doses producing 50% antagonism (ED50) of the pancuronium-induced neuromuscular block were 0.50, 0.54 and 0.71 mg/kg for LF-14, 2,4-DAP and 4-AP respectively. The compounds had only slight cardiovascular effects. In contrast to 4-AP, LF-14 and 2,4-DAP did not reduce the duration of ketamine/diazepam induced anesthesia, suggesting minimal if any central nervous system effects of these two compounds. PMID- 3034627 TI - GABAA receptors in the midbrain central grey mediate the antiaversive action of GABA. AB - Intracerebral injection of the GABAA agonists muscimol (1 nmol), isoguvacine (1 nmol) or THIP (1, 2 and 4 nmol) in rats with chemitrodes implanted in the dorsal midbrain central grey raised the threshold electrical current for inducing escape behaviour. The effect of THIP was dose-dependent. In contrast, the GABAB agonist baclofen (10 and 100 nmol) did not affect the aversive threshold. Furthermore, pretreatment with baclofen (10 nmol and 100 nmol) did not significantly change the effect of THIP (2 nmol). These results indicate that the antiaversive action of GABA in the midbrain central grey is mediated by GABAA but not by GABAB receptors. PMID- 3034628 TI - Benzodiazepine and beta-carboline modulation of GABA-stimulated 36Cl-influx in cultured spinal cord neurons. AB - GABAA agonists stimulate 36Cl-influx in spinal cord cultured neurons in a concentration-dependent manner. This effect of GABAA receptor stimulation is enhanced by benzodiazepines like clonazepam, diazepam and flurazepam and attenuated by (+)bicuculline and picrotoxinin. The beta-carbolines, methyl-6, 7 dimethoxy-4-ethyl-beta-carboline-3-carboxylate (DMCM) and propyl-beta-carboline-3 carboxylate (beta-CCPr) exhibited opposite effects, with DMCM attenuating, while beta-CCPr potentiating GABA's effect. These results are consistent with the behavioral and electrophysiological effect of benzodiazepines and beta-carbolines with GABA receptor complex. PMID- 3034629 TI - Increase of peripheral type benzodiazepine binding sites in kidney and olfactory bulb in acutely stressed rats. AB - Fifteen minutes after the initiation of swimming stress in the rat we observed a 50% increase in the number of [3H]RO 5-4864 binding sites in kidney and a 37% increase in the olfactory bulb, without change in affinity. The binding in heart and cerebral cortex remained unchanged after the stress. These results are discussed in relation to previous work on both the action of an acute stress in central benzodiazepine receptors and the possible modulation of peripheral benzodiazepine receptors of the kidney by adrenocortical hormones. PMID- 3034630 TI - The novel anticonvulsant MK-801 interacts with central phencyclidine recognition sites in rat brain. PMID- 3034631 TI - Kappa-opiate-induced diuresis and changes in blood pressure: demonstration of receptor stereoselectivity using (+)- and (-)-tifluadom. AB - (+)-Tifluadom injected i.p. produced a biphasic response of urine output: within the first hour of its administration the drug produced antidiuresis followed by a diuretic phase. In contrast, the (-) isomer produced a modest reduction in urine output as compared to the output of the saline-treated rats. In addition, (+) tifluadom markedly reduced the output of urinary Na+ and K+. The effects of (+) tifluadom were blocked by 7.5 mg/kg naloxone but not by 10 mg/kg of the benzodiazepine antagonist Ro 15-1788. Parallel experiments demonstrated that the i.v. administration of (+)-tifluadom to non-anesthetized rats caused a dose related pressor response that lasted for at least 15 min. This effect of (+) tifluadom was blocked and antagonized by naloxone. In contrast, (-)-tifluadom was either inactive on the cardiovascular system or produced short-lasting hypotension. In pentobarbital-anesthetized rats, 100 micrograms/kg (+)-tifluadom caused a precipitous hypotension that was reversed by naloxone but not by Ro 15 1788. PMID- 3034632 TI - Effect of mouse peritoneal macrophages of orally administered very high dilutions of silica. AB - The activity of very high dilutions of silica, a substance cytotoxic for macrophages, was tested on the synthesis by mouse peritoneal macrophages of the inflammatory ether-lipid paf-acether and its precursor lyso paf-acether. C57Bl6 female mice received for 25 days either 1.66 X 10(-11) M silica (11 sil) or 1.66 X 10(-19) M (19 sil) (final concentration) in the tap-water they were given to drink while control mice remained untreated. Isolated macrophages from mice treated with 11 sil produced 44.2 and 30.8% more paf-acether than cells from untreated mice in the presence of 50 and 200 micrograms zymosan (Z)/ml respectively. When 19 sil was given to the mice, the respective increases were 67.5 and 38%. In an experiment with a blind design, the mice were either untreated or received 19 sil or saline submitted to the same dilution procedure (19 sal). After administration of 19 sil, paf-acether synthesis was 55.5 and 33.5% higher upon stimulation with 50 and 200 micrograms Z/ml, respectively, than in the 19 sal group. In a third blind experiment, macrophages from mice that received 19 sil formed 61.3 and 28.6% more paf-acether upon stimulation with 50 and 200 micrograms Z/ml respectively, as compared to mice receiving 19 sal or lactose submitted to the same dilution procedure (19 lac). There was no difference between the 19 sal and the 19 lac groups. The differences between control and silica-treated mice were highly statistically significant in all experiments. There was no effect on the synthesis of lyso paf-acether. These results demonstrate clear ex vivo cellular effect of high dilutions of silica, that cannot be explained in our present state of knowledge. PMID- 3034634 TI - Characteristics of sodium-induced increase in opiate antagonist binding sites in rat brain membranes. AB - Sodium decreases agonist binding and increases antagonist binding to opiate receptor sites in brain membranes. This study characterizes in detail the 20-40% increase in [3H]naloxone binding caused by sodium. This increase in binding was specific to sodium (not mimicked by potassium or lithium) and was maximal at 10 mM NaCl. The sodium effect was reversible: washing membranes free of sodium restored binding to normal. Sodium increased Bmax, not KD, of [3H]naloxone binding. The sodium-induced increased binding was not inhibited by either N ethylmaleimide (NEM) or phospholipase A2 at concentrations which inhibit 50-85% of normal [3H]naloxone binding. In NEM-treated membranes, the effect of sodium on increasing naloxone binding was actually increased. The regional distribution of these sodium-dependent sites were different from normal naloxone sites. These results suggest that physiological concentrations of sodium expose naloxone sites which differ in biochemical properties from naloxone sites assayed in the absence of sodium. PMID- 3034633 TI - Effects of synthetic omega-conotoxin on synaptic transmission. AB - The effects of chemically synthesized omega-conotoxin GVIA (a neurotoxic peptide from Conus geographus) on synaptic transmission at the bullfrog sympathetic ganglion, frog neuromuscular junction and electric organ of the ray, Narke japonica, were studied. The synthetic toxin irreversibly suppressed synaptic transmission at these synapses by arresting the release of transmission from the nerve terminals without showing postsynaptic effects. This action of the toxin was effectively antagonized by high concentrations of extracellular Ca2+. The synthetic toxin irreversibly blocked the Ca2+-dependent action potential of bullfrog sympathetic ganglion cells. These results suggest that omega-conotoxin GVIA blocks synaptic transmission by interfering with the Ca2+ influx through the voltage-sensitive Ca2+ channel of the nerve terminal. These results indicate that the chemically synthesized omega-conotoxin GVIA acts exactly like the natural omega-conotoxin GVIA. Thus, the synthetic toxin can be used in place of the natural toxin as a useful probe for the voltage-sensitive Ca2+ channel in the nervous system. PMID- 3034636 TI - Effects of drugs affecting noradrenergic neurotransmission in rats with spontaneous petit mal-like seizures. AB - Wistar rats of a strain displaying spontaneous petit mal-like seizures and spike wave EEG discharged (SWD) were injected i.p. with drugs affecting noradrenergic neurotransmission. The EEG and behavior were recorded. Drugs which decrease alpha noradrenergic neurotransmission, prazosin (alpha 1-antagonist) and clonidine (alpha 2-agonist), increased SWD and were sedative in a dose-dependent manner. Drugs which increase alpha-noradrenergic neurotransmission, ST 587, cirazoline (alpha 1-agonists) and yohimbine (alpha 2-antagonist), reduced SWD and the latter two caused agitation. Drugs which interact with beta-noradrenergic transmission (salbutamol, isoprenaline and propranolol), monoamine oxidase inhibitors (nialamide and iproniazid), and a noradrenaline reuptake inhibitor (desipramine), did not affect SWD. These findings suggest that noradrenaline participates in the control of petit mal-like seizures in the rat, as in other types of seizures and other animal models. PMID- 3034635 TI - Potentiation of inflammatory reactions in guinea-pig skin by an angiotensin converting enzyme inhibitor (MK 422). AB - There have recently been reports of persistent cough and increased broncho obstruction likely to have been induced by ACE inhibitors. In order to study the effect of MK 422 (the active parent diacid of enalapril) on the inflammatory response, ovalbumin-sensitized guinea-pigs were tested intradermally with ovalbumin, capsaicin and bradykinin. All inflammatory responses were enhanced by treatment with MK 422 for 2 days prior to testing as compared to the responses of control animals. Infiltration of neutrophils, eosinophils, basophils and monocytes was increased following ovalbumin challenge in the MK 422-treated animals. We suggest that skin reactions and airway symptoms noticed during ACE inhibitor therapy might have been due to induced or potentiated inflammatory reactions in the skin or in the bronchial wall. PMID- 3034637 TI - Behavioural evidence for an interdependence between GABAA receptors and beta 2 adrenoceptors. AB - The possibility of a functional interdependence between central GABAA receptors and beta 2-adrenoceptors has been investigated using the ability of both types of agonist to potentiate the tic (head-twitch) response to 5-methoxy-N,N dimethyltryptamine in the mouse. At a dose which selectively antagonised beta 1- but not beta 2-adrenoceptors, ICI 118,551 abolished the effects of single doses of muscimol, diazepam and pentobarbitone. Conversely, bicuculline abolished the potentiation caused by the beta 2-adrenoceptor agonist procaterol. Thus there is preliminary evidence that these two receptor types do show mutual interdependence. PMID- 3034639 TI - Effect of acidosis on alpha 1- and alpha 2-adrenoceptor-mediated vasoconstrictor responses in isolated arteries. AB - We have investigated the effect of reducing the pH (from 7.5 to 7.0 by addition of HCl) on vasoconstrictor responses to noradrenaline in cat middle cerebral artery (in which responses are mediated almost entirely by alpha 2-adrenoceptors) and in rabbit pulmonary artery (in which responses are mediated by alpha 1 adrenoceptors). In the cerebral artery, a reduction in pH caused a pronounced inhibition of the responses to noradrenaline, and the antagonistic effect of idazoxan (100 nM) was increased 10-fold. In contrast, in the pulmonary artery, a reduction in pH had no effect on the responses to noradrenaline and the antagonistic effect of prazosin (100 nM) was not altered. We conclude that acidosis selectively reduces the vasoconstriction mediated by alpha 2 adrenoceptors in vitro. PMID- 3034638 TI - Down-regulation of atrial natriuretic peptide receptor and cyclic GMP response in cultured rat vascular smooth muscle cells. AB - Treatment of cultured rat vascular smooth muscle cells with human atrial natriuretic peptide (hANP) or Met(O)12hANP caused a similar and marked reduction (approximately 80%) of ANP receptor number (down-regulation). A second challenge with hANP stimulated the accumulation of intracellular cGMP in the down-regulated cells to the same extent as in control cells. These data suggest that ANP receptor sites are functionally heterogenous, the more abundant site being uncoupled from guanylate cyclase but susceptible to down-regulation. PMID- 3034641 TI - Mediastinal germ cell tumor in trisomy 8. PMID- 3034640 TI - Electrophysiological evidence that Ro 15-4513 is a benzodiazepine receptor inverse agonist. PMID- 3034642 TI - Small cell lung cancer. AB - It can be stated that, depending on the type of lung cancer, the best opportunity for curative treatment is in the early stage of disease, when cancer is limited to the lung and surgical intervention can be indicated. Especially in the case of SCLC, the number of patients presenting with such a limited disease is very low. In SCLC, chemo- and/or radiotherapy can induce initial good responses, which, although not curative in most cases, can elongate life for an average of 10 months. Although changes and refinements in treatment are continuously introduced, further progresses in the outcome have not achieved for the last decennium. Therefore, more fundamental research is needed to indicate new treatment avenues. A number of findings appear promising in this field. Firstly, the development of tissue culture techniques has enabled a better study of the biological properties of both SCLC and non-SCLC. Secondly, the development of monoclonal antibodies has refined the possibilities to type lung cancer. Particularly if monoclonal antibodies could be identified which occurrence turns out to be relevant to prognosis, immunohistopathology could become an important additional tool for the assessment of a pathological diagnosis in lung cancer. In addition, monoclonal antibodies which are specific for lung cancer could be used for an immunotherapeutical approach in the near future. Such a treatment might complement currently available treatment modalities. PMID- 3034643 TI - Small cell lung cancer. 27th annual meeting of the Netherlands Federation of Medical Societies. Groningen, April 4, 1986. Proceedings. PMID- 3034644 TI - Intermediate filament expression in small cell lung cancer; poor correlation to in vitro data. AB - A panel of monoclonal antibodies, detecting different intermediate sized filament proteins, was prospectively applied on all specimens derived from S.C.L.C. patients attending our clinic in 1985. Reactivity with the antibodies was subsequently correlated to clinical data. The results indicate a heterogeneous pattern of reactivity of the assessed antibodies. However this heterogeneity is not a straightforward extension of the intermediate sized filament expression in SCLC cell lines. PMID- 3034645 TI - Genome analysis of small cell lung cancer (SCLC) and clinical significance. AB - A chromosome analysis of three cell lines derived from SCLC showed deletions of the short arm of chromosome 3 with bands p21-p23 as the shortest region of overlap. Hybridization of a polymorphic 3p21 probe to DNA from leukocytes of seven SCLC patients revealed heterozygosity for two of them. In the tumours of both these patients the probe detected homozygosity. This suggests the presence of a mutant cancer gene in the short arm of chromosome 3 which might express itself and/or activate some oncogene(s) after deletion of a suppressing normal allele. Amplification of the oncogene C-MYC was found in four cell lines including the ones cytogenetically analyzed. Amplification of C-MYC, though to a lesser degree, was also found in an available pleural effusate from which one of these lines had been established. As shown by in situ hybridization, the amplified oncogene was present in double minutes in three of the cell lines. In the remaining line it was in a homogeneously staining chromosome region. All patients from whom cell lines with C-MYC amplification were obtained had a negative response to chemotherapy. The observed correlation between amplification of C-MYC, occurrence of so-called variant type SCLC-derived cell lines, and negative response to chemotherapy indicates that a genome analysis of SCLC might provide further criteria for the characterization and subdivision of this highly malignant cancer and thereby a base for an optimal selection of therapy for distinct cases of SCLC. PMID- 3034646 TI - Doubling time of neuron-specific enolase and survival in small cell lung cancer patients. Results of a preliminary analysis. AB - During a retrospective analysis of the value of neuron specific enolase (NSE) in patients with small cell lung cancer (SCLC) it became apparent that at progressive disease (PD) NSE rose exponentially with a doubling time (NSE-Td) varying from 10 - 94 days. In this study the influence of the NSE-Td on the survival of 29 SCLC-patients has been investigated. A significant correlation between survival from the start of rise of NSE at PD and NSE-Td was observed. By extrapolating the exponential rise of NSE to the start of treatment a theoretical logarithmic value of NSE, called Yr, could be calculated. When the patients were grouped according to the Yr value greater than -1, between -1 and -4 and less than or equal to -4 a highly significant correlation between the survival from the start of treatment and NSE-Td was found in all 3 groups. These preliminary data suggest that by means of NSE-Td and Yr value the survival of an SCLC-patient from the time of rise of NSE and from the start of treatment may be predicted within certain limits. PMID- 3034647 TI - Heterogeneity of lung cancer: the problem of sample error in diagnostic electron microscopy. AB - We studied the ultrastructure of superficial and deep samples of 40 resected primary lung carcinomas. Tumour cell differentiation was semiquantitatively assessed and differences between samples of a same tumour were evaluated. In two instances were major differences in ultrastructural diagnosis found between the samples of the same tumour. A further 9 cases showed one predominant differentiation in one sample, but two equally predominant differentiations in the second sample. The other 29 tumours did show occasional minor differences between the samples, but these differences did not result in differences in ultrastructural diagnosis. PMID- 3034648 TI - Clinical implications of the biology of small cell lung cancer. AB - Considerable advances have been made over the past 5 years of our understanding of the biology of lung tumours in particular SCLC. In this chapter, and in other sections of this symposium various aspects of these properties have been discussed. The greater understanding of the properties of these tumours should provide means whereby increased success can be achieved in clinical remissions and long term survival for the majority of patients with these diseases. PMID- 3034649 TI - Activity of (7) anthracycline related compounds in an doxorubicin sensitive human small cell lung cancer line and its doxorubicin resistant descendant. Activity of doxorubicin, daunorubicin, 4-deoxyrubicin, 4-demethoxydaunorubicin, detorubicin, 4'-epidoxorubicin and mitoxantrone. PMID- 3034651 TI - Preliminary results of the pathological review of a small cell lung cancer trial (EORTC 08825). PMID- 3034650 TI - Small cell lung cancer trials in the EORTC. AB - An overview of activities of the EORTC Lung Cancer Cooperative Group in small cell lung cancer is given. Optimal length of therapy, identification of patients with good/bad prognostic factors, role of surgery and the study of new drugs presently are the main areas of interest for the group. With its large patient potential the group is able to verify "early favourable reports" and to quickly perform phase II studies. PMID- 3034652 TI - High-dose etoposide for central nervous system metastases of small cell lung cancer. Preliminary results. AB - High-dose etoposide (1.0-1.5 g/m2) was given to 17 small cell lung cancer (SCLC) patients with metastases in the central nervous system. In 4 out of 9 evaluable patients with brain metastases and 4 out of 5 patients with meningeal carcinomatosis a response was seen. In all patients severe myelosuppression was observed. Three patients died of septicemia during the aplastic phase. Despite severe toxicity high-dose etoposide is potentially useful for CNS metastases of SCLC. PMID- 3034654 TI - Hypnotic action of pentobarbital in mice: a possible mechanism. AB - The effect of GABA receptor agonists (THIP and baclofen) on the hypnotic activity (loss of righting reflex and sleep time) of pentobarbital in mice was investigated. Combinations of either THIP-pentobarbital or baclofen-pentobarbital interacted synergistically by increasing hypnotic activity. The GABAa receptor antagonist, bicuculline, decreased the hypnotic effect of THIP-pentobarbital combinations but not that of baclofen-pentobarbital combinations. These results suggest that the hypnotic activity of pentobarbital involves GABAa receptor function. PMID- 3034653 TI - Ultrastructure of the gracile nucleus projection to the dorsal accessory subdivision of the cat inferior olive. AB - This study examined the termination pattern within the dorsal accessory subdivision of the cat inferior olive of axons arising from the gracile nucleus. The gracile terminals were labeled by anterograde transport of wheat germ agglutinin complexed to horseradish peroxidase and visualized with tetramethyl benzidine. Gracile terminals were found to contain round synaptic vesicles and form asymmetric synaptic contacts. Of particular interest was the finding that gracile axons, like axons from the spinal cord, terminate primarily outside of synaptic glomeruli. Yet most of the gracile terminals did not synapse on isolated dendritic elements. Rather, the majority contacted distal dendrites which directly contacted other dendritic elements, forming simple complexes termed dendritic thickets. Typically the dendritic thickets were composed of two or three dendrites that received input from more than one round vesicle-containing synaptic terminal. Only one terminal per thicket was labeled by injections in the gracile nucleus. This clustering of pre- and postsynaptic elements within the thickets provides opportunities for many of the same interactions allowed by synaptic glomeruli, in particular divergence and convergence of information. PMID- 3034655 TI - Toxoplasma gondii and Hammondia hammondi: DNA comparison using cloned rRNA gene probes. AB - A mung bean nuclease genomic library of purified DNA from tachyzoites of the RH strain of Toxoplasma gondii was prepared in the bacteriophage lambda gtll and recombinants containing rRNA gene fragments were detected by hybridization with radiolabeled total RNA from the closely related coccidian Eimeria acervulina. Ten recombinants were chosen at random, and five of these were investigated further using probes for the genes of the large and small rRNA of Plasmodium berghei. An insert (called TG4) that hybridized only to the 3' end of the large rRNA coding region of P. berghei and an insert (called TG18) that hybridized only to the small rRNA coding region of P. berghei were purified by electrophoresis in low melting point agarose. Radiolabeled E. acervulina total RNA, TG4, and TG18, were then used to compare the sizes of the large and small rRNA gene fragments after DNA extracted from three strains of T. gondii, and the type strain of the closely related coccidian Hammondia hammondi were cut by one of a series of 10 restriction endonucleases. The patterns obtained for the three T. gondii isolates were identical to those obtained for H. hammondi, for each enzyme tested. In addition, the guanine plus cytosine (G + C) content of H. hammondi DNA was found to be almost identical to that obtained previously for T. gondii DNA. PMID- 3034656 TI - Taenia hydatigena: isolation of mitochondrial DNA, molecular cloning, and physical mitochondrial genome mapping. AB - Mitochondrial DNA was isolated from Taenia hydatigena, T. crassiceps, and Echinococcus granulosus using a cetyltrimethylammonium bromide precipitation technique. The technique is simple, rapid, reproducible, and does not require extensive high speed ultracentrifugation. The advantage of using mitochondrial DNA from taeniid cestodes for comparative restriction analysis was demonstrated. Mitochondrial DNA of T. hydatigena was isolated as covalently closed circular molecules. These were linearized by single digestion with BamHI and the molecular weight was estimated from the linear form of 17.6 kb. The mitochondrial DNA of T. hydatigena is therefore similar in size and structure to that of many other animal species. The entire mitochondrial genome was cloned into pBR322 in Escherichia coli and a restriction map of the recombinant molecule was constructed. The potential of using the cloned mitochondrial genome as a probe in speciation studies as well as for providing functional information on the role of the cestode mitochondrion is discussed. PMID- 3034657 TI - Ascaris suum: partial isolation and characterization of hypodermis from the adult female. AB - A method was developed to remove the muscle from body wall strips of adult female Ascaris suum resulting in a hypodermis cuticle preparation. Optimum treatment for obtaining the hypodermis cuticle was a 15 min incubation with trypsin (2.0 mg/ml) at room temperature, followed by mechanical removal of the muscle. The hypodermis cuticle prepared in this manner incorporated radiolabeled amino acids into cuticular and hypodermal proteins; incorporation was inhibited by protein synthesis inhibitors. Characterization of the hypodermal proteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that the hypodermis apparently contains proteins that differ from those of the cuticle and that the hypodermis of adult A. suum appears to lack cuticle protein precursors. This result will now allow detailed biochemical and physiological investigations of the hypodermis, a tissue which is critical for cuticle synthesis. PMID- 3034658 TI - Na, K ATPase activity during early postnatal development of the rat submandibular gland. AB - The activity of the ouabain-sensitive Na, K ATPase was measured in membrane fractions of the submandibular gland of 1-, 7-, 14- and 21-day-old rats. This activity increased with age and reached adult levels by 21 days. PMID- 3034659 TI - Effect of phorbol myristate acetate on cultured tumor cells derived from different stages of avian sarcoma virus (ASV)-induced neoplastic growth. AB - We have investigated the action of the tumor promoter phorbol 12-myristate-13 acetate (PMA) on avian sarcoma cells cultured from avian sarcoma virus (ASV) induced tumors at various stages of growth and on normal chicken embryo fibroblasts (CEF). We found that exposure to PMA (30 ng/ml) led to decreased expression of the malignant phenotype of avian sarcoma cells which had been derived from progressively-growing or regressing neoplasms. For example, treatment with PMA caused inhibition of growth of cultured avian tumor cells as well as decreased synthesis of DNA and protein. In addition, such treatment resulted in reduced expression of pp60src kinase activity. Direct inoculation of PMA into growing avian sarcomas led to an enhancement of the tumor regression which occurs in this system as a consequence of a strong cell-mediated, anti tumor immune response. PMID- 3034661 TI - A rosette receptor assay with haem-microbeads. Demonstration of a haem receptor on K562 cells. AB - A rosette assay was developed for the detection of haem receptor-bearing cells. Indicator particles were prepared by covalent binding of haem to acrylic microbeads. The new method was tested on K562 human erythroleukaemia cells, known to take up haem. In tests on several batches, 80-90% of the K562 cells were rosetted with haem-microbeads whereas mature erythrocytes were haem receptor negative. Rosette formation was inhibited in a dose-dependent manner by micromolar concentrations of free haemin but not by albumin or transferrin. Uncoated microbeads or albumin-coated microbeads did not attach to K562 cells but transferrin-coated microbeads rosetted 50-70% of them. Diferric transferrin inhibited these rosettes, but haemin had no effect. PMID- 3034660 TI - Field trial evaluation of an inactivated rotavirus vaccine against neonatal diarrhea of calves. AB - Field trials were conducted using an inactivated rotavirus vaccine for prevention of calf neonatal diarrhea. For the trials, 458 pregnant cows from 26 herds were involved. In each herd, cows which had been inseminated within a period of two months were selected and randomly subdivided in two groups. Cows in one group (248 head in total) were vaccinated 6 weeks before calving and again 4 weeks later; cows in the other group (210 head in total) were left as unvaccinated controls. At calving, colostrum was collected from each cow and stored at -30 degrees C until used for feeding calves. The newborn calves, beginning the second day of life and for the next 7-10 consecutive days, each was fed a daily supplement of 400 ml of colostrum from its dam. The diarrhea occurred in 86 (40.9%) calves that had received colostrum from unvaccinated dams (normal colostrum), and in 7 (2.8%) calves which were fed colostrum from vaccinated dams (immune colostrum). The disease was very severe in the normal colostrum-fed calves and 52 of them died. Those calves which survived the disease underwent a significant loss of condition. By contrast, the 7 immune colostrum-fed calves displayed a rather mild enteric condition, and all recovered without any sequela being observed. PMID- 3034662 TI - Expression of CD 25 (Tac antigen) in lymphoid leukemias and non-Hodgkin lymphomas. AB - A monoclonal antibody to IL2 receptor Tac/CD 25 antigen was included in the phenotyping panel of 92 leukemic proliferations. Spontaneous expression of this molecule was observed in one disorder of T-phenotype only, but in 75% of B chronic lymphocytic leukemia (CLL) and on cells from B-cell leukemic lymphomas. These findings provide further evidence for a non-T-cell restriction of IL2 receptor and raise the question of multiple gene derepression in hematologic disorders of B-lineage. PMID- 3034665 TI - Control of cytoplasmic pH by Na+/H+ exchange in rat peritoneal macrophages activated with phorbol ester. AB - The mechanisms underlying cytoplasmic pH (pHi) regulation in elicited rat peritoneal macrophages were investigated by electronic sizing and fluorescence determinations. Acid-loaded cells rapidly regained normal pHi by means of an amiloride-sensitive Na+/H+ exchange. When stimulated by 12-O-tetradecanoyl phorbol 13-acetate, macrophages displayed a biphasic pHi change: a marginal acidification followed by an alkalinization. The latter results from activation of Na+/H+ exchange, since it is Na+-dependent and prevented by amiloride. When the antiport is inhibited, the full magnitude of the initial acidification can be appreciated. This acidification is independent of the nature of the ionic composition of the medium and probably reflects accumulation of protons generated during the metabolic burst. Under physiological conditions, these protons are rapidly extruded by the Na+/H+ antiport. PMID- 3034663 TI - (3H)ouabain binding to leukaemic cells and intralymphocytic sodium content in chronic lymphocytic leukaemia; no evidence for alterations of the Na+/K+-pump. AB - The number of specific (3H)ouabain binding sites and dissociation constants (Kd) were determined by Scatchard analysis of values for leucocytes from patients with B-cell chronic lymphocytic leukaemia (CLL), chronic myeloid leukaemia (CML), acute blastic leukaemia (AL) and healthy subjects. CLL lymphocytes and normal B cells bound significantly less (3H)ouabain than did normal T-lymphocytes. CML granulocytes showed the same binding characteristics as normal granulocytes, while blast cells from AL patients bound significantly more (3H)ouabain than did normal granulocytes or B-cells. The increased binding capacity in blast cells might, at least partly, reflect their larger cell size. A decrease in Kd values was only found in CLL lymphocytes, as compared with normal B-cells. Intralymphocytic sodium content in CLL lymphocytes was significantly increased, as compared with that in T-cell-enriched normal lymphocytes. (3H)ouabain binding did not show any relationship to different prognostic variables in CLL. The present data mainly argue against altered Na+/K+-ATPase enzyme activity as an indicator of malignancy. PMID- 3034666 TI - A fragment of an endogenous inhibitor produced in Escherichia coli for calcium activated neutral protease (CANP) retains an inhibitory activity. AB - A C-terminal fragment of an endogenous rabbit liver inhibitor for calcium activated neutral protease (CANP) was produced in Escherichia coli and its inhibitory activity was examined after purification. The truncated inhibitor (373 amino acid residues), which contains two internal repeat structures, inhibits 2 mol CANP whereas the native liver inhibitor (639 residues), containing four internal repeat structures, inhibits 4 mol CANP. This supports the hypothesis that the repeating unit is the functional unit of inhibition. The results also indicate that post-translational modification of the inhibitor is not essential for inhibition. PMID- 3034664 TI - [Effect of cardiac glycosides on the calcium ion binding by biomembrane lipids]. AB - By using methods of fluorescent and spine (electron paramagnetic resonance) probes it was shown that cardiac glycosides (digoxin, digitoxin, convallatoxin, corelborine and strophanthin) effectively interact with biomembrane lipids enhancing conformational motility of lipids and cause additional binding of calcium ions with the membrane. PMID- 3034667 TI - Differential effects of DNA tumor virus nuclear oncogene products on adipocyte differentiation. AB - We have introduced SV40 and polyoma large T antigen- and adenovirus-type 12 E1A genes into mouse 3T3-L1 preadipocyte cells to study the ability of various nuclear oncogene products to modulate cell differentiation. Clones expressing E1A products could differentiate into adipocytes faster than the control in spite of the absence of adipogenic inducers, as measured by the appearance of lipid droplets microscopically and by staining accumulated triglycerides with oil red O. However, clones expressing SV40 and polyoma large T antigens could not differentiate even if they were exposed to the inducers. PMID- 3034668 TI - Enkephalin-degrading enzymes and angiotensin-converting enzyme in human and rat meninges. AB - The neutral endopeptidase NEP 24.11 (enkephalinase) has been visualized in human spinal cord by in vitro autoradiography using [3H]HACBO-Gly as a radiolabelled probe. The specific binding was present in the substantia gelatinosa and particularly dense in meninges surrounding the spinal cord. Enzymatic studies using [3H][D-Ala2, Leu]enkephalin as substrate confirmed the presence of NEP in dura and pia mater of human tissue. In addition, the human meninges were shown to contain high concentrations of angiotensin-converting enzyme (ACE) and aminopeptidases. The three enzymes have also been detected in rat tissues but their distribution pattern differs from that of human tissue. In dura mater, 45% of the [Leu]enkephalin hydrolysis was due to enkephalinase and 38% to bestatin sensitive aminopeptidases. In contrast in pia mater aminopeptidases were more efficient in hydrolyzing enkephalin. The possible role of these enzymes in the meninges could be to maintain the homeostatic concentration of neuropeptides in the central nervous system. PMID- 3034669 TI - Regional distribution of leukotriene and mono-hydroxyeicosatetraenoic acid production in the rat brain. Highest leukotriene C4 formation in the hypothalamus. AB - The regional distribution of ionophore A23187-induced synthesis of leukotrienes and mono-hydroxyeicosatetraenoic acids in the rat brain in vitro was investigated. Pronounced differences in leukotriene C4 formation were observed, with the highest synthetic capacity in the hypothalamus. The formation of leukotriene C4 was about 12-times higher in the hypothalamus as compared to the cerebellum. This finding is in agreement with a possible neuroendocrine role for leukotriene C4. In contrast, the activity of leukotriene B4 synthesis was widely distributed without pronounced regional differences in the rat brain. Formation of 5-, 9-, 11-, 12- and 15-monohydroxyeicosatetraeonoic acid was detected in all regions. The major lipoxygenase product in the hypothalamus and thalamus was 5 hydroxyeicosatetraenoic acid, while other monohydroxyeicosatetraenoic acids predominated in the remaining regions tested. PMID- 3034670 TI - Counterion collapse and the effect of diamines on bacteriorhodopsin. AB - A recent report of electrical measurements on oriented bacteriorhodopsin in gels [(1986) FEBS Lett. 195, 164 168] concluded that low concentrations of diamines reversed the direction of the proton pump. Calculations are presented which show that in low diamine concentrations, charge displacements of the counterion atmosphere in the direction opposite to proton pumping are expected following H+ ejection. It is also shown that the effect will be sharply reduced by raising the diamine concentration or by adding excess salt, as was observed. Hence it is not necessary to conclude that diamines reverse the direction of the proton pump itself. PMID- 3034671 TI - Association of calpains 1 and 2 with protein kinase C activities. AB - Calpains 1 and 2 co-eluted with protein kinase C activities after hydrophobic (phenyl-Sepharose) and anion-exchange (Mono Q) chromatographies of a 100,000 X g supernatant which was defined as cytosol. After centrifugation of the cytosol at 200,000 X g for 16 h, the major part of calpain 1 and of its associated protein kinase C activity was recovered in the pellet, when the major part of calpain 2, also associated to a protein kinase C activity, was present in the resulting supernatant. Polyacrylamide gel electrophoresis of the fractions eluted from the Mono Q column, which contained calpains 1 or 2 and their associated protein kinase C activities, revealed two main bands with a molecular mass of 80 and 28 kDa. PMID- 3034673 TI - Inhibitory effects of pertussis toxin on a depolarization-evoked Ca2+ influx in NG108-15 cells. AB - Depolarized stimulation 1.5-fold increased Ca2+ influx which was inhibited by pretreatment with verapamil or LaCl3. Treatment with pertussis toxin, islet activating protein (IAP), induced a reduction in 50 mM K+-induced Ca2+ influx and stimulated adenylate cyclase (AC) activity in NG108-15 cells. However, addition of dibutyryl cAMP or forskolin treatment elevating cAMP level exerted no effects on a depolarization-induced Ca2+ influx. Dissociated B-oligomer of IAP after treatment with dithiothreitol and ATP increased a depolarization-evoked Ca2+ influx. It is suggested that inhibitory GTP-binding protein (G1) or other IAP substrate proteins could directly be involved in Ca2+ influx via voltage sensitive Ca2+ channel. PMID- 3034672 TI - The rate of oxygen consumption and superoxide anion formation by stimulated human neutrophils. The effect of particle concentration and size. AB - The respiratory burst of neutrophils was measured as a function of the ratio of the opsonised beads to neutrophils. The rate of oxygen uptake was found to be linear up to a bead:neutrophil ratio of 25. The production of the superoxide anion, as measured by the rate of reduction of cytochrome c, was negligible until a certain 'critical' value of the bead:neutrophil ratio was reached. The rate of oxygen consumption and superoxide release above the critical value varies linearly with the bead:neutrophil ratio. Both the rate of oxygen consumption and of superoxide release vary with the square of the radius of the ingested particle. It is suggested that this depends on the surface area of neutrophil membrane, activated by contact with the antagonist. PMID- 3034675 TI - In situ distribution of EcoRI methylase and restriction endonuclease in cells of Escherichia coli Bs 5. AB - Specific IgG antibodies were raised in rabbits against purified EcoRI methylase and restriction endonuclease. Post embedding labeling experiments, using the protein A-gold technique, were made with paraformaldehyde-glutaraldehyde fixed cells, embedded in Lowicryl K4M resin at low temperatures. Labeling with methylase-specific antibodies showed 60-70% of gold particles in the cytoplasm and 30-40% at the cell envelope, whereas the use of restriction enzyme-specific antibodies led to a distribution of 10-30% in the cytoplasm and 70-90% in the cell envelope. The results coincide with the proposed function of the enzymes: in the cytoplasm methylase protects the cells' own DNA from self-destruction, and the restriction endonuclease cuts foreign DNA when entering the cell. PMID- 3034674 TI - Insulin stimulates a novel GTPase activity in human platelets. AB - Insulin stimulated the activity of a high-affinity GTPase activity in human platelet membranes some 62% over that of the basal activity. Half-maximal stimulation (Ka) was achieved with 3.1 nM insulin. The Km for GTP of the insulin stimulated GTPase was 0.6 microM GTP. Treatment of isolated platelet membranes with cholera toxin, but not pertussis toxin, blocked insulin's ability to stimulate GTPase activity. Cholera toxin acted as a more potent inhibitor of the insulin-stimulated GTPase activity than that of the GTPase activity of the stimulatory guanine nucleotide regulatory protein, Gs, as monitored by stimulation using prostaglandin E1 (PGE1). Mixed ligand experiments showed that insulin stimulated GTPase activity in an additive fashion to GTPase activity stimulated by PGE1, due to Gs; by adrenaline (+ propranolol), due to the inhibitory guanine nucleotide regulatory protein, G1 and by vasopressin, which stimulates the putative 'Gp', a G-protein suggested to control the stimulation of inositol phospholipid metabolism. Insulin thus appears to stimulate a novel high affinity GTPase activity in human platelet membranes. This may reflect the functioning of the putative Gins, a guanine nucleotide regulatory protein which has been suggested to mediate certain of insulin's actions on target tissues. PMID- 3034676 TI - Absorption mode two-dimensional NOE spectroscopy of exchangeable protons in oligonucleotides. AB - A new NMR method is described for the generation of absorption mode two dimensional NOE spectra of oligonucleotides in H2O solution. The method yields spectra that are free of baseline distortions with excellent suppression of the intense H2O resonance. The method is demonstrated for a sample of the dodecamer d(CGCGAATTCGCG)2. All exchangeable base protons are identified and a number of new types of NOE connectivities are observed. PMID- 3034678 TI - Glucose-induced degradation of yeast fructose-1,6-bisphosphatase requires additional triggering events besides protein phosphorylation. AB - Glucose addition to yeast cells stimulates a cAMP overshoot with concomitant activation of cAMP-dependent protein kinase, which in turn rapidly phosphorylates fructose-1,6-bisphosphatase. The phosphorylated enzyme subsequently undergoes a slow proteolytic breakdown. Also, it has been proposed that phosphorylation represents the mechanism that initiates proteolysis. Here we present experiments carried out on a yeast mutant defective in adenylate cyclase [(1982) Proc. Natl. Acad. Sci. USA 79, 2355-2359] in which extracellular cAMP triggers full enzyme phosphorylation but a scanty proteolysis, whereas glucose plus cAMP provoke both phosphorylation and complete proteolytic breakdown. Thus, besides a glucose induced cAMP peak, which results in enzyme phosphorylation, other effects evoked by the sugar are indispensable for its proteolytic degradation. PMID- 3034677 TI - Is enhanced free radical flux associated with increased intracellular proteolysis? AB - Intracellular proteolysis was measured in cultured cells during and after free radical attack. Radicals were generated firstly, throughout the aqueous phase by gamma irradiation and secondly, selectively, either extracellularly or intracellularly by chemical and enzymic methods. With both approaches, stimulation of proteolysis was observed in certain circumstances. Phenylhydrazine stimulated proteolysis at low concentration but inhibited at higher. Depletion of the antioxidant glutathione and inhibition of catalase also increased proteolysis. PMID- 3034679 TI - On the dephosphorylation of the ATP,Mg-dependent protein phosphatase modulator. AB - The dephosphorylation of the modulator subunit is an essential step in the kinase FA-mediated activation of the ATP,Mg-dependent protein phosphatase. Mg2+ is implicated in this autocatalytic dephosphorylation which is not effected by the addition of phosphoinhibitor-1. Dephosphorylation of free modulator by the catalytic subunit is also largely Mg2+-dependent but can be abolished by phosphoinhibitor-1 in concentrations comparable to the amount of modulator used as substrate (micromolar). The phosphorylase phosphatase activity of the catalytic subunit is inhibited by nanomolar concentrations of phosphoinhibitor-1 and is completely independent of divalent cations. PMID- 3034680 TI - [Enzymatic isolation of the pyramidal neurons of the rat hippocampus]. PMID- 3034681 TI - [Action of nicotinamide on neuromuscular transmission]. PMID- 3034683 TI - [Effect of acetylcholine and carbamylcholine on the resting membrane potential of denervated muscle in the rat]. AB - The decrease of resting MP after denervation in diaphragm muscle of the rat was studied in vitro. Transitory activation of Na+,K+-pump by carbamylcholine, acetylcholine or adrenaline hyperpolarized the muscle fiber membrane but did not compensate the postdenervation fall of the MP. Similarly, the permanent presence of carbamylcholine at culturing media did not prevent the development of changes in the MP after denervation. The decrease of MP was accompanied by spontaneous release of acetylcholine for 24 hrs. Synaptic acetylcholine seems to play no obvious role in neurotrophic control of the resting MP. PMID- 3034682 TI - [Opiate receptors of the brain]. PMID- 3034684 TI - [Variations in the sensitivity of the neuromuscular junctions of the frog to the blocking action of hyperkalemic solutions]. AB - The depression of evoked transmitter release with solutions with enhanced (up to 8 mM) potassium concentration was studied in cut and voltage-clamped muscle fibers of frog m. cutaneous pectoris. As potassium blockade proceeds, a reduction of the second end-plate current to paired stimuli occurs before a change in the first end-plate current, the reduction of end-plate currents being simultaneous with the latency increase. The existence of variable sensitivity of different end plates to potassium blocking action depends on year season and ionic environment. Possible mechanisms of potassium blocking action are discussed. PMID- 3034685 TI - [Participation of metabolic systems of carotid chemoreceptors in their effector regulation]. AB - Electrical stimulation of the rat sinus nerve altered fluorescence of restored pyridin nucleotides and oxidative flavoproteids in the carotid body cells due to enhancement of cellular respiration. This was followed by an increase of cGMP content in the carotid body. The greatest shifts of these parameters occurred in square-wave stimulation with 20/sec frequency. The effector influences on the carotid body metabolism are most effectively blocked with pentamine and seem to be of cholinergic nature, the transmitter effect of acetylcholine being modulated with enkephalins. The cGMP system may serve as an intermediary of the effector influences. PMID- 3034686 TI - [Effect of blockade and stimulation of adrenoreceptors on the pumping function of the heart in animals adapted and unadapted to physical loading]. AB - Experiments with blockade of beta-adrenoceptors in rats revealed chronotropic effects of adrenaline and noradrenaline, as well as action of noradrenaline on systolic blood volume. The influence of adrenaline on systolic blood volume increases in that case, whereas combined blockade of both types of adrenoceptors sharply lowers it. In case of separate blockade of alpha-adrenoceptors, the action of adrenaline changes slightly. In physical exercise, adaptation of chronotropic and systolic effects of catecholamines are changed reciprocally. In condition of beta-adrenoceptor blockade and prior to it, the influence of adrenaline on systolic blood volume is much higher in adapted animals than in the control ones. PMID- 3034687 TI - [Regional changes in circulation in response to stimulation of beta-adrenergic receptors with isoproterenol]. AB - The muscle and adrenal fractions of cardiac output increased after injection of isoproterenol. The relative blood flow increase in skeletal muscles was accompanied with the increase in regional by-pass blood flow. The latter or reduction in the vessel permeability were found in the lungs. The cardiac output fractions decreased in extensive areas of skin, brain, myocardium, internal organs of hepatic portal vein, kidneys, testicles. The signs of the blood flow increase were found in the brain, myocardium, liver. The relative blood volume decreased in the majority of internal organs and tissues while increasing in the liver. PMID- 3034688 TI - [Condylomata acuminatum of the oral cavity]. PMID- 3034689 TI - Paget's disease in an epidermal cyst. AB - A case of Paget's disease involving both the epidermis of the nipple and an adjacent epidermal cyst is reported. Immunohistochemistry showed similarity between the infiltrating cells in both sites and those of the underlying mammary adenocarcinoma. Epidermal cysts may be involved in a variety of disease processes which affect the epidermis in general and, unless they are subjected to careful histopathological examination, these diseases may be overlooked. PMID- 3034691 TI - Plasma beta-endorphin, beta-lipotropin and corticotropin in polycystic ovarian disease. AB - In 9 women with polycystic ovarian disease (PCOD) and in 11 control subjects at the follicular phase of the normal cycle, blood samples were collected at 15-min intervals during a 2 h period of bed rest for the assay of beta-endorphin, beta lipotropin, corticotropin, cortisol and prolactin. During the study period, the plasma levels of these hormones decreased more significantly in the PCOD than in the control group, suggesting that the PCOD patients had a more significant stress response to the puncture of the vein than the control subjects. The second hour of the study period was considered to represent resting levels of hormones. The mean resting levels (+/- S.E.) of the hormones between the PCOD and control groups, respectively, were as follows: beta-E, 2.0 +/- 0.4 vs. 1.1 +/- 0.1 pmol/l, p less than 0.05; beta-LPH, 3.4 +/- 0.6 vs. 2.1 +/- 0.5 pmol/l, N.S.; corticotropin, 2.0 +/- 0.3 vs. 1.1 +/- 0.5 pmol/l, p less than 0.05; cortisol, 176 +/- 24 vs. 128 +/- 16, N.S.; and prolactin; 3.9 +/- 0.6 vs. 5.6 +/- 1.2 ng/ml, N.S. These results confirm the previous findings on increased circulating levels of beta-E in PCOD. A concomitant increase of the plasma level of corticotropin suggests that the basal secretion of both beta-E and corticotropin from the anterior pituitary gland is increased in women with PCOD. PMID- 3034690 TI - Response of psoriatic lesions to multiple applications of leukotriene B4 and 12 HETE. AB - Single topical application of the neutrophil chemoattractant arachidonic acid metabolites leukotriene B4 (LTB4) and 12-R,S-hydroxy-5,8,10,14-eicosatetraenoic acid [corrected] (12-HETE) to normal skin elicits a histological response similar to early psoriasis. This effect diminishes following repeated application indicating the development of local tolerance. In order to assess whether induction of tolerance could be exploited as a treatment for psoriasis we studied the clinical effects of topical application of LTB4 (n = 6) and 12-HETE (n = 3) under occlusion to small psoriatic lesions for 12 consecutive days. The area of the control lesions decreased significantly over the study period while the areas of lesions to which LTB4 and 12-HETE were applied remained unchanged. No other difference between responses of the three groups was detected. We have shown that, in the doses used, multiple applications of these substances to established stable psoriatic lesions do not produce a therapeutically useful response. PMID- 3034693 TI - 'High-affinity' peripheral-type benzodiazepine-binding sites. PMID- 3034692 TI - Receptor-mediated endocytosis in steroid hormone-producing tissue. PMID- 3034694 TI - Increase in liver protein phosphatase-1 in spontaneously diabetic Chinese hamsters. AB - Two broad-specifically protein phosphatases, termed protein phosphatase-1 (PrP-1) and protein phosphatase-2A (PrP-2A), accounting for all the hepatic activity regulating glycogen phosphorylase, were measured in spontaneously diabetic Chinese hamsters exhibiting persistent glycosuria. When compared with genetically related inbred sublines free of glycosuria, diabetic animals demonstrated approximately 25% increase in PrP-1 activity measured either in crude tissue extracts or in cytosols fractionated by ion-exchange chromatography. No significant alteration in total PrP-2A activity was observed in the diabetic animals. These findings indicate that a specific change in hepatic PrP-1 is associated with genetically acquired diabetes in Chinese hamsters. In contrast to reported data using animals with experimentally induced diabetes mellitus, hepatic PrP-1 was increased in the spontaneously diabetic Chinese hamsters. The data suggests that distinct alterations in PrP-1 and associated metabolic consequences are exhibited by different types of diabetes. PMID- 3034695 TI - Sensitization of adrenocortical cell adenylate cyclase activity to ACTH by angiotensin II and activators of protein kinase C. AB - Exposure of bovine adrenocortical cells to optimal concentrations of angiotensin II (A II) resulted in an almost 2-fold enhancement of cellular cAMP accumulation in response to steroidogenic concentrations of ACTH. This effect was dose dependent and transient, with a maximum after 4-6 min of treatment with A II. Activators of protein kinase C such as 12-O-tetradecanoyl-phorbol-13-acetate (TPA) and 1,2-dioctanoyl-sn-glycerol mimicked that effect in a sustained fashion. The ACTH-sensitized state of the adrenocortical adenylate cyclase system induced by TPA exhibited also an enhanced response to forskolin. On the other hand, previous treatment of the cells by pertussis toxin suppressed any further effect of TPA. It is suggested that, following A II exposure, the Gi inhibitory components of the adrenocortical cell adenylate cyclase system may be inactivated, leading to increased response to ACTH. This process may involve protein kinase C activation, subsequent to intracellular generation of lipidic messengers resulting from accelerated phosphoinositide breakdown induced by angiotensin. PMID- 3034696 TI - Catecholestrogens stimulate progestin secretion by cultured porcine granulosa cells. AB - The enzymatic metabolism of estradiol (E2) to the catecholestrogens, 2 hydroxyestradiol (2-OH-E2) and 4-hydroxyestradiol (4-OH-E2) in granulosa cells has been reported. Therefore, we evaluated the effects of these compounds and compared them to those of E2 on porcine granulosa cells cultured in serum-free medium. Cultures of granulosa cells were exposed to various treatments of E2, 2 OH-E2, 4-OH-E2 and(or) follicle-stimulating hormone (FSH) for 4 days and concentrations of progesterone in medium and cell numbers were determined. After 4 days of treatment, 2-OH-E2 and 4-OH-E2 stimulated basal progesterone production by granulosa cells, but 4-OH-E2 was less effective than 2-OH-E2. 2-OH-E2 (1 microgram/ml) stimulated progesterone production by 3.3 +/- 0.6-fold (n = 6 experiments), whereas E2 (1 microgram/ml) stimulated progesterone production 9.9 +/- 1.7-fold (n = 6 experiments). 2-OH-E2 at 4 micrograms/ml further stimulated progesterone production to 10.7 +/- 2.2-fold above controls (n = 9 experiments), whereas 4 micrograms/ml of E2 did not cause further stimulation of progesterone production. Thus, the average potency of 2-OH-E2 was less than E2. Concurrent treatment with 2-OH-E2 (4 micrograms/ml) and saturating concentrations of E2 resulted in further significant increases in progesterone production above the effects of either single treatment both in the absence and presence of FSH (200 ng/ml).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3034698 TI - In vitro effects of adenohypophysial hormones on rat pineal melatonin content and release. AB - The effect of adenohypophysial hormones on rat pineal melatonin content and release was examined in vitro. Medium concentration of radioimmunoassayable melatonin decreased after a 6 h exposure to 1-100 ng/ml FSH; pineal levels of melatonin were only decreased by 100 ng/ml FSH. LH (1-100 ng/ml) augmented significantly medium melatonin concentration, tissue levels being increased at 10 ng/ml LH. Parallel increases of explant and medium melatonin content were found after exposure to 1-100 ng/ml TSH. At the smallest concentration employed (1 ng/ml) prolactin increased melatonin content and release while at 100 ng/ml a significant depression of both parameters was found. Growth hormone (1-10 ng/ml) augmented melatonin levels in medium but failed to modify them at 100 ng/ml, although at this concentration tissue melatonin levels increased. ACTH did not modify pineal melatonin synthesis in vitro. PMID- 3034697 TI - High density lipoprotein influences the response of rat adrenocortical cells to gamma 3-melanotropin. AB - Gamma 3-melanotropin (gamma 3-MSH) exhibits a marked dose-dependent synergism with ACTH1-24 on corticosterone production by cells isolated from the inner zones of the rat adrenal cortex. This phenomenon is demonstrated to best advantage when donor animals are killed after stress or pretreatment with ACTH. If adrenal cells are prepared from quiescent or hypophysectomized animals, the inclusion of high density lipoprotein (HDL) in the incubation medium is required for a significant gamma 3-MSH response. Dibutyryl cAMP can successfully substitute for ACTH1-24 in these incubations but rat low density lipoprotein does not reproduce the HDL effect. These data are consistent with our in vivo studies demonstrating that gamma 3-MSH potentiation is a product of increased cholesterol mobilization within the adrenal cortex and suggest that in the rat, a significant source of the cholesteryl ester pool which is responsive to gamma 3-MSH may derive from circulating HDL. PMID- 3034699 TI - Role of second messengers in the regulation of glucagon secretion from isolated rat islets of Langerhans. AB - The roles of diacylglycerol (DAG), cAMP and Ca2+ in mediating the stimulatory action of arginine on pancreatic A cells have been investigated using phorbol esters, forskolin, a Ca2+ ionophore and trifluoroperazine (TFP). 0.5 microM 4 beta-phorbol 12-myristate 13-acetate (PMA) which stimulated glucagon secretion by approximately 3-fold in the absence of arginine, was unable to enhance arginine stimulated glucagon secretion. Higher concentrations (1 and 10 microM) of PMA were able to enhance glucagon secretion in the presence of 1.25, 2.5, 5 but not 10 mM arginine. Insulin secretion was enhanced by PMA under all the conditions tested. Arginine (10 and 20 mM)-stimulated secretion of glucagon and insulin were synergistically augmented by 20 microM forskolin. While the effects of forskolin plus PMA on the A cells were additive, the effects of the two agents on the B cells were synergistic. The responses of the A and B cells to arginine required extracellular Ca2+. Secretion of the two hormones was dose-dependently stimulated by A23187. Arginine-stimulated glucagon secretion but not insulin secretion, was dose-dependently inhibited by TFP. These results suggest that proposed cellular second messengers interact differently in the A and B cells, and DAG and Ca2+ may play pivotal roles in mediating the actions of arginine on the A but not B cells; cAMP may play a modulatory role in the A cell response to arginine. PMID- 3034700 TI - Starfish oocyte maturation: 1-methyladenine triggers a drop of cAMP concentration related to the hormone-dependent period. AB - Oocyte maturation (meiosis reinitiation) in starfish is induced by the natural hormone 1-methyladenine (1-MeAde). Oocytes of Evasterias troschelii contain 0.43 pmole cyclic AMP/mg protein and 0.47 pmole cyclic GMP/mg protein. Upon stimulation by 1-MeAde the oocytes undergo a moderate (10-30%) decrease in their cAMP concentration. The concentration of cGMP remains unaltered. Oocytes treated with forskolin, an activator of adenylate cyclase, increase their cAMP concentration over 35-fold, up to 16 pmole cAMP/mg protein. When stimulated by 1 MeAde these forskolin-pretreated oocytes undergo a major (50-70%) decrease in their cAMP concentration. A similar decrease is triggered by mimetics of 1-MeAde, such as dithiothreitol, arachidonic acid (AA), and 8-hydroxyeicosatetraenoic acid (8-HETE), but not by adenine which is inactive. 1-MeAde-stimulated oocytes of Pisaster ochraceus also undergo a decrease in cAMP content, the size of which is increased by forskolin. Although a decrease in cAMP begins at sub-threshold 1 MeAde concentrations, the maximal decrease occurs at the same concentration of 1 MeAde needed for maturation induction and a further 1000-fold increase of the 1 MeAde concentration has no further effect. Upon removal of 1-MeAde, the cAMP concentration immediately increases to its original level. Sequential addition and removal of 1-MeAde triggers a sequential decrease and increase of the cAMP concentration, illustrating the continuous requirement for 1-MeAde for eliciting the decrease. Successive additions of 1-MeAde, however, do not trigger further decreases of the cAMP concentration. The temperature dependences of the cAMP concentration decrease and of the hormone-dependent period (HDP; the time of contact with 1-MeAde required for induction of maturation) are closely related. Forskolin, which increases the cAMP concentration, also increases the duration of the HDP (2.5-fold), delays the time course of protein phosphorylation burst and germinal vesicle breakdown, and inhibits AA- and 8-HETE-induced maturation. We conclude that 1-MeAde triggers a drop in cAMP concentration, which is tightly associated with the hormone-dependent period of oocyte maturation. PMID- 3034701 TI - Distribution of cAMP-dependent protein kinase during development in Dictyostelium discoideum. AB - During the developmental cycle of Dictyostelium discoideum cyclic AMP functions as both a chemotactic signal for aggregation and a regulatory molecule during later events of differentiation. Morphological and biochemical data suggest that cAMP may direct cells during morphogenesis and differentiation. We utilized microtechniques to determine the stage- and cell-specific levels of the cAMP dependent protein kinase, the probable intracellular cAMP receptor. Kinase activity was low and non-cAMP-dependent in amoebae and early aggregates but increased and became cAMP-dependent in aggregates after the formation of tight cell contacts. Maximum kinase activity and cAMP dependency occurred during the slug and culmination stages. The only differential distribution of the kinase within a single stage occurred during culmination when the activity in the stalks was approximately one-fourth of that in the prespore mass. Preliminary evidence indicates that this difference is not due to an inhibitor. In all other stages tested cAMP-dependent protein kinase activity was equal in prespore and prestalk cells. PMID- 3034702 TI - Two Drosophila learning mutants, dunce and rutabaga, provide evidence of a maternal role for cAMP on embryogenesis. AB - The dunce gene of Drosophila melanogaster encodes a cAMP-specific phosphodiesterase (form II). Mutant dunce flies have elevated levels of cAMP and exhibit a number of defects including learning deficiencies and female sterility. Two partial suppressors of the female sterility phenotype have been selected in an X chromosome containing a dunce null mutation. Both suppressors are associated with reduced AC2 activity. Complementation analyses suggest that both are alleles of the learning mutant rutabaga. Females homozygous for dunce null mutations that abolish PDE activity do not deposit eggs. The suppressors exhibit differential effects on egg deposition and production of progeny; double-mutant females deposit many eggs that fail to hatch, but some develop to adults. These adult progeny exhibit morphological defects that are confined mostly to the second and third thoracic segments or to the first five abdominal segments. These observations demonstrate that the dunce gene is required in adult females for egg laying and that the dunce gene provides an essential maternal function required for normal development of the zygote. Clonal analysis, employing the dominant female-sterile mutation ovoD1, demonstrates that the former requirement for PDE activity resides in somatic cells and that the latter requirement resides in germ line cells. Female germ line cells homozygous for a dunce null mutation produce oocytes that fail to develop. Thus, homozygous dunce null-mutant zygotes develop to adults solely because of the enzyme or mRNA present in the oocytes of heterozygous mothers. Mutant alleles of rutabaga act in the germ line cells to partially suppress the developmental defects caused by dunce mutations. Thus the rutabaga gene, as well as the dunce gene, functions in both somatic and germ line cells. PMID- 3034704 TI - Strategies of foreign gene expression in cultured animal cells. PMID- 3034703 TI - Differential effects of activators of cAMP-dependent protein kinase and protein kinase C on cleavage of one-cell mouse embryos and protein synthesis and phosphorylation in one- and two-cell embryos. AB - Membrane-permeable cAMP analogs or elevation of intracellular cAMP by cyclic nucleotide phosphodiesterase (PDE) inhibitors activates cAMP-dependent protein kinase. Biologically active phorbol esters or diacylglycerol activate the calcium , phospholipid-dependent protein kinase, protein kinase C (PK-C). We report that membrane-permeable cAMP analogs, PDE inhibitors, biologically active phorbol esters, or a synthetic diacylglycerol inhibited cleavage of 1-cell mouse embryos to the 2-cell stage. The cAMP analogs and PDE inhibitors were effective only when added prior to S of the first cell cycle, whereas PK-C activators inhibited cleavage when added up until late G2/M. The PDE inhibitor Ro 20 1724/1 inhibited both DNA and protein synthesis in 1-cell embryos, whereas the phorbol ester, 12-O tetradecanoyl-phorbol-13 acetate, or alpha-amanitin did not. In addition, 1-cell embryos prevented from cleaving by PDE inhibitors did not show specific changes in the pattern of protein phosphorylation associated with the 2-cell embryo, whereas such changes occurred in 1-cell embryos inhibited from cleaving with PK-C activators. Transcription in the 2-cell embryo results in the synthesis of a specific set of proteins, which is inhibited by alpha-amanitin. Although treatment of 1-cell embryos with aphidicolin or PK-C activators during G1 did not inhibit the synthesis of these proteins, treatment with cAMP analogs or PDE inhibitors during G1 inhibited the appearance of these proteins. These results are discussed in terms of how the synthesis of transcription-dependent proteins in the 2-cell embryo may be regulated by protein phosphorylation. PMID- 3034706 TI - The growth of polio virus in human diploid fibroblasts grown with cellulose microcarriers in suspension cultures. AB - Human diploid fibroblasts have been successfully grown with cellulose fiber microcarriers in suspension giving values of between 1 to 2 X 10(6) cells/ml. When infected with polio virus type 1 titers were obtained which were at least as good or better than those obtained with monkey kidney cells and similar to that obtained with Vero cells. Good results were also obtained using conventional microcarriers such as Cytodex, Biosilon and Gelibeads. Our results suggest that many of the technical difficulties in growing human diploid fibroblasts on microcarriers can be overcome allowing these cells to be a favourable alternative to heteroploid cells as a substrate for viral vaccine production. PMID- 3034705 TI - Present knowledge of poliovirus and perspectives for future vaccines. AB - Will currently used poliovirus vaccines be improved or be replaced with new ones? By analyzing recent data on poliovirus antigenicity and on the molecular basis of its attenuation, we have tried to answer this question. PMID- 3034707 TI - Cell type dependence of herpes simplex virus gene expression and of processing of viral protein. AB - R325, a recombinant virus constructed from the wild type herpes simplex virus 1 strain F [HSV-1(F)] carries a 500 bp deletion in the alpha 22 gene. Both viruses are temperature sensitive for growth and late gene expression in HEp-2 cells. To determine the properties of alpha 22 protein and to identify the product of the non deleted 5' portion of the alpha 22 gene in R325, infected BHK and HEp-2 cells incubated at permissive and non-permissive temperatures were electrophoretically separated in denaturing polyacrylamide gels, electrically transferred to a nitrocellulose sheet and reacted with rabbit polyclonal antibody to an oligopeptide synthesized according to the predicted aminoacid sequence of the 5' terminus of alpha 22. The results were as follows: the authentic alpha 22 protein was detected in infected BHK and HEp-2 cells maintained at either 34 degrees C (permissive) or 39 degrees C (non-permissive) temperatures, the truncated protein specified by R325 accumulated at 39 degrees C but not at 34 degrees C. Furthermore, the proteins made in BHK cells were processed to higher apparent molecular weights and formed several bands in contrast to the single band formed by the protein made in HEp-2 cells; the temporal pattern of expression of HSV 1(F) and R325 at the 39 degrees C was more advanced in BHK cells than in HEp-2 cells. Specifically, in BHK cells at the non-permissive temperature alpha protein synthesis was shut off earlier and gamma proteins not made in infected HEp-2 cells were detected. These studies suggest that folding of viral proteins may be in part cell dependent and either compensate or exacerbate the effects of mutations in specific genes. PMID- 3034708 TI - Multiple variants in foot-and-mouse disease virus (FMDV) populations: the Achilles heel for peptide and rec. DNA vaccines? AB - Variants of type A10 FMDV were isolated by passage of virus in BHK-cells in the presence of a neutralizing anti-peptide serum or monoclonal antibodies. These variants which were no longer neutralized by the particular anti-peptide serum or monoclonal antibody were easily obtained from (crude) virus populations ("cattle" virus and BHK-adapted virus). The rapidity of isolation (in two or three passages) suggested that these variants are already present in normal virus populations. All (plaque purified) variants isolated so far seem to be different: A panel of 20 monoclonal antibodies and an anti-peptide serum showed different neutralization patterns for all isolates and parent virus. Electrofocusing patterns of many variants were found to be different showing changed charges for VP2 as well as for VP1. Thus in FMDV both VP1 and VP2 are probably involved in antigenic sites. Normally the variants escape our attention because in neutralization assays only a limited quantity of infective units are used, representing only the "top of the iceberg". In classical inactivated virus vaccines many of these variants will be represented and therefore can be expected to be "primed" immunogenetically. This will not be the case with peptide vaccines or in case of recombinant DNA products, where only one virus clone is represented. In addition, and probably more important, inactivated virus vaccines will raise antibodies against completely independent epitopes that each have a limited chance to be changed in the variants present in the challenge virus population. Thus if peptide vaccines can be composed in such a way that antibodies are raised against completely different antigenic sites the chance of break through of variants will be strongly limited and it is expected that the efficient protection of inactivated virus vaccines can be approached. PMID- 3034709 TI - Challenging settled opinions in classic foot-and-mouth disease vaccine preparation. AB - In a previous study we challenged the generally accepted opinion that inactivation of FMDV by formaldehyde (FA) is an (unsafe) non-linear process. Our data showed that under proper conditions inactivation will be linear without "tailing-off". For more than forty years two other fixed beliefs existed with respect to FMD vaccine preparation: Virus must first be adsorbed to Al(OH)3-gel before being inactivated. Concentrations of formaldehyde are critical and must be within a very narrow range. Until recently the prescription of adsorption of the virus prior to inactivation made proper control of inactivation kinetics impossible. However, by eluting the virus from the gel by caesium chloride density centrifugation, the kinetics of inactivation can be studied. For adsorbed and non-adsorbed virus, identical inactivation curves were found. Antigenicity was found to be of identical "quality" for adsorbed and non-adsorbed virus. The second dogma was challenged by inactivating non-adsorbed virus with FA concentrations of up to 6 times the one originally prescribed. In parallel inactivation was performed with acetyl-ethylene-immine (AEI). The antigen was purified after inactivation. Antigen yields after purification were systematically lower at high FA-concentrations, however, antigenicity seemed only slightly changed by the treatments: In an immunosorbent (ELISA) assay using a panel of 22 monoclonal antibodies (McAb) only few McAb's reacted differently with FA-treated antigen if compared with AEI-treated or fresh (A10) FMDV. The FA treatment induced decreases, as well as increases in reactivities. The AEI treated virus reacted almost identically to non-treated 146S particles.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3034710 TI - Diabetes decreases Na+-K+ pump concentration in skeletal muscles, heart ventricular muscle, and peripheral nerves of rat. AB - Na+-K+-ATPase or the Na+-K+ pump is essential for some specific properties of muscle and nerve tissue such as contractility and excitability. Previous studies have shown conflicting variations in Na+-K+-ATPase activity or Na+-K+ pump concentration of muscle cells in experimental diabetes. Our study demonstrates that early untreated diabetes in rats induced by injection of streptozocin is associated with decreases in [3H]ouabain binding-site concentration of 24-48% in various skeletal muscles and 16% in peripheral nerves as well as a decrease in K+ dependent 3-O-methylfluorescein phosphatase activity of 21% in the heart ventricle. These effects could be prevented by insulin treatment. They probably represent a decrease in the concentration of Na+-K+ pumps. There was no evidence for more than one population of Na+-K+ pumps in intact samples of skeletal muscle and nerves from normal, diabetic, and insulin-treated animals. The decrease in Na+-K+ pump concentration in nerve cells may be due to atrophy of the axons. In skeletal muscles, myocardium, and peripheral nerves, the observed decrease in Na+ K+ pump concentration may be important for the pathophysiology of diabetes. We emphasize that quantification of Na+-K+-ATPase or the Na+-K+ pump in muscle and nerve tissue from diabetic animals should preferably be performed with either intact samples or crude homogenates of whole tissue. PMID- 3034712 TI - Reduction of the observed prevalence of so-called non-A non-B hepatitis using sensitive markers of HBV and herpes viruses infections. AB - In the absence of a specific marker, the observed prevalence of so called non-A non-B hepatitis depends on the sensitivity of the markers of the other viral infections known to induce hepatitis. We have reevaluated this prevalence after using sensitive markers of HBV (HBs monoclonal radioimmunoassay M-RIA and IgM anti-HBc), EBV (IgM anti-VCA), CMV (IgM anti-CMV) and HSV (IgM anti-HSV) in a group of 53 subjects usually considered as having acute or chronic hepatitis. Detection of IgM against HBc, CMV and HSV used immunocapture tests. Among the 37 patients with acute hepatitis, 11 (30 p. 100) were positive for at least one sensitive marker, including 10 markers of HBV (7 M-RIA and 3 IgM anti-HBc) and one IgM anti-CMV. Among the 16 patients with chronic hepatitis, one was positive for HBV by M-RIA; five patients had a false positive reaction to EBV (IgM anti VCA) disappearing when rheumatoid factor was eliminated. This study shows that many cases of the so-called non-A non-B hepatitis are in fact due to HBV or to a variant of HBV. Definition of non-A non-B hepatitis must include subjects negative for HBV by M-RIA and IgM anti-HBc and negative for CMV by IgM anti-CMV. PMID- 3034713 TI - [Treatment of 25 patients with hepatocellular carcinoma with an anti-androgen, cyproterone acetate (Androcur)]. PMID- 3034711 TI - The onset of liver glycogen synthesis in fasted-refed lean and genetically obese (fa/fa) rats. AB - Lean and genetically obese (fa/fa) rats were fed ad libitum, or fasted for 17 h and then meal-fed for varying time intervals. During refeeding, glucose-6 phosphatase activity of lean rats declined to the low value that was present in livers of fasted obese rats and which remained unchanged in the obese group during the meal. Refeeding also resulted in increases in hepatic concentrations of glucose-6-phosphate and fructose-6-phosphate, fructose 1,6-bisphosphate, fructose-2,6-bisphosphate, alpha-glycerophosphate, pyruvate and lactate in lean and obese rats, absolute values being higher in the fasted obese than in the fasted lean group. Obese animals had higher postprandial portal blood insulin, glucose and lactate concentrations than lean animals. In spite of this, the rate of hepatic glycogen deposition was the same in both groups and was accompanied by similar glycogen synthase a levels. Following refeeding, phosphorylase was transiently inactivated in livers of lean but not of obese animals, while glycogen synthase was inactivated in both groups. The data suggest that in lean animals refeeding was associated with a stimulation of liver glycolysis, presumably by insulin; in fasted obese rats hepatic glycolysis was already in a stimulated state and was only slightly enhanced further after the meal, in keeping with their unaltered hyperinsulinaemia; there was an increased turnover of liver glycogen or a resistance to insulin stimulation of glycogen synthesis in fa/fa rats during refeeding. PMID- 3034714 TI - Diffuse hepatocellular dysplasia and carcinoma associated with the Mmalton variant of alpha 1-antitrypsin. AB - The cirrhosis and hepatocellular carcinoma associated with alpha 1-antitrypsin deficiency has been exclusively reported with the PI Z allele. We present a 63-yr old white man with emphysema, cirrhosis, and hepatocellular carcinoma. The latter occurred on a background of diffusely distributed hepatocellular dysplasia. Serum protein electrophoresis suggested a deficiency of alpha 1-antitrypsin quantitated at 13% of normal. PI phenotyping showed that he had only the rare PI Mmalton allele, previously associated only with severe lung disease. Family studies demonstrated the distribution of this rare allele. The liver at autopsy displayed well-differentiated hepatocellular carcinoma in addition to alpha 1-antitrypsin deposits in normal, dysplastic, and malignant cells. PMID- 3034715 TI - The effects of forskolin, cAMP, and cyanoketone on steroid-induced meiotic maturation of yellow perch (Perca flavescens) oocytes in vitro. AB - Intact yellow perch (Perca flavescens) follicles stimulated by 17 alpha, 20 beta dihydroxy-4-pregnen-3-one (17 alpha, 20 beta-PG) to undergo germinal vesicle breakdown (GVBD) in vitro were incubated with several agents which have been shown to increase cellular cAMP levels. Two phosphodiesterase inhibitors, SQ20,006 and isobutyl-methyl-xanthine, blocked GVBD at 1.0 mM. At lower levels (0.5, 0.1 mM) there was a dose-response effect and SQ20,006 was more inhibitory. Forskolin at 1.0-20.0 microM blocked steroid-induced GVBD, but levels of 0.1 microM or less were noninhibitory. In time-course experiments, significant inhibition of GVBD was observed when SQ20,006 (1.0 mM) was added within 6 hr after steroid stimulation or forskolin (10.0 microM) was added within 12 hr. When SQ20,006 was administered in 6-hr pulses and then removed, inhibition was observed only when the steroid was given as a 1-hr prepulse which was removed at the start of the incubation period. In this case, GVBD was blocked if the SQ20,006 pulse was given before 18 hr. At 10.0 mM, cAMP completely inhibited GVBD but was noninhibitory at lower levels. However, lower levels of cAMP (1.0, 0.5 mM) and forskolin (0.1 microM) were inhibitory if the follicles were also incubated with 1.0 microgram/ml of cyanoketone, an inhibitor of steroidogenesis. These results indicate that in vitro, increases in cAMP are inhibitory to steroid induced meiotic maturation but may stimulate steroidogenesis in the follicle wall as well. Furthermore, in vitro steroid-stimulated maturation can be inhibited by increased cAMP for a relatively long time, following steroid treatment. PMID- 3034716 TI - In vitro effect of prostaglandins on the accumulation of cyclic AMP in the avian oviduct. AB - The effects of prostaglandins E1, E2, and F2 alpha (PGE1, PGE2, and PGF2 alpha, respectively) on cyclic AMP production in tissue samples from the uterus (U), uterovaginal sphincter (UVS), and vagina (V) of regularly laying hens as well as the effects of PGE2 and forskolin on U and V of Japanese quail were studied. Both PGE1 and PGE2 enhanced cyclic AMP production in the hen oviductal segments, particularly in the V in a dose-related manner, PGE1 being the more effective agonist. PGF2 alpha had no significant effect. Quail U appeared to be less responsive to PGE2 which caused a 40% increase in cyclic AMP levels at the highest concentration tested (28 microM), whereas forskolin at 10 microM increased cyclic AMP production in quail U and V 7- and 20-fold, respectively. It is suggested that the primary function of PGE2 and PGE1 is to relax the terminal portion of the oviduct, allowing the expulsion of the egg under the contractile influence of PGF2 alpha and other oxytocic factors. PMID- 3034717 TI - Mode of replicon fusion mediated by the duplicated insertion sequence IS21 in Escherichia coli. AB - The insertion sequence IS21 (2.1 kb) originating from the broad-host-range IncP plasmid R68 transposes infrequently; by contrast, the IS21 tandem repeat found on the derivative R68.45 is highly active in transpositional mobilization of other replicons in a variety of Gram-negative bacteria. The mobilized plasmids are joined to R68.45 by single IS21 copies in direct orientation. The formation of IS21 tandem duplications was observed in cointegrates between R68.45 and pBR325::IS21 and also in an RP1::IS21 plasmid derivative in which a segment located between two directly repeated copies of IS21 was deleted spontaneously. We speculate that IS21 tandem repeats can arise when the termini of two IS21 elements are specifically joined in a transposition or deletion event. A resistance gene flanked by two IS21 elements in direct orientation did not behave as a transposon. The omega fragment carrying transcription and translation stop signals was inserted into various sites of the IS21 tandem repeat; in this way it could be shown that the left IS21 element (which is next to the kanamycin resistance gene in R68.45) was 100 times more active in cointegrate formation than was the right-hand element. Cointegrates between the conjugative plasmid R751 and pBR325 derivatives carrying IS21 and IS21::omega in tandem contained a single IS21 at one replicon junction and a single IS21::omega at the other. In the IS21 duplications the inner IS21 ends were preferentially recognized (presumably by IS21 transposase), whereas the outer termini were not required for cointegrate formation. Based on these findings a conservative (simple) pathway of transposition is proposed for R68.45 and other plasmids with an IS21 tandem repeat.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3034719 TI - Three genes are required for trans-activation of Ty transcription in yeast. AB - Mutations in the SPT3 gene were isolated as one class of suppressors of Ty and solo delta insertion mutations in Saccharomyces cerevisiae. Previous work has shown that null mutations in SPT3 abolish the normal Ty delta-delta transcript; instead, a transcript that initiates 800 bases farther downstream is made, suggesting that SPT3 is required for transcription initiation in delta sequences. We have selected for new spt mutations and have screened for those with the unique suppression pattern of spt3 mutations with respect to two insertion mutations. Our selection and screen has identified two additional genes, SPT7 and SPT8, that are also required for transcription initiation in delta sequences. We show that mutations in SPT7 or SPT8 result in the same alteration of Ty transcription as do mutations in SPT3. In addition, mutations in all three genes cause a sporulation defect. By assay of a Ty-lacZ fusion we have shown that spt3, spt7 and spt8 mutations reduce transcription from a delta sequence by 10-25-fold. Finally, we show that SPT3 mRNA levels are unaffected in either spt7 or spt8 mutants, suggesting that these two genes do not regulate transcription of SPT3. PMID- 3034718 TI - PET111, a Saccharomyces cerevisiae nuclear gene required for translation of the mitochondrial mRNA encoding cytochrome c oxidase subunit II. AB - Mutations in the nuclear gene PET111 are recessive and specifically block accumulation of cytochrome c oxidase subunit II (coxII), the product of a mitochondrial gene. However, the coxII mRNA is present in pet111 mutants at a level approximately one-third that of wild type. The simplest explanation for this phenotype is that PET111 is required for translation of the coxII mRNA. The reduced steady-state level of this mRNA is probably a secondary effect, caused by increased degradation of the untranslated transcript. Mitochondrial suppressors of pet111, carried on rho-mtDNAs, bypass the requirement for PET111 in coxII translation. Three suppressors are fusions between the coxII structural gene and other mitochondrial genes, that encode chimeric proteins consisting of the N terminal portions of other mitochondrially coded proteins fused to the coxII precursor protein. When present together with rho+ mtDNA in a heteroplasmic state, these suppressors allow coxII synthesis in pet111 mutants. Thus in wild type, the PET111 product, or something under its control, probably acts at a site coded in the proximal portion of the gene for coxII to promote translation of the mRNA. PET111 was isolated by molecular cloning and genetically mapped to a position approximately midway between rna1 and SUP8 on chromosome XIII. PMID- 3034720 TI - Genetic transformation of Drosophila melanogaster with an autonomous P element: phenotypic and molecular analyses of long-established transformed lines. AB - Following transformation of a Drosophila melanogaster true M strain with an autonomous P element, six lines were established and monitored for their molecular and phenotypic properties during a 4-yr period. The number of P elements increased with time in all the lines but the rate of increase differed among lines. Furthermore, degenerate elements arose in each of the lines during propagation. By the end of the 4th yr, the total number of elements in every line was similar to that of a very strong P strain.--At the phenotypic level, all of the transformed lines evolved high P activity, but only three developed complete or nearly complete regulatory ability. The other three lines attained only intermediate levels of regulation over the 4-yr period. One of these lines was particularly noteworthy. Although it contained as many as 55 P elements per genome (20 of which were potentially complete) and had extremely high P activity potential, it continued to exhibit limited regulatory ability. In addition, when females of this line were maintained at high temperatures, the ability to suppress P activity was even further diminished. A strain with this combination of molecular and phenotypic properties, in an apparently stable configuration, has not been previously described.--The results are discussed in the context of the possible role of degenerate elements in regulating P element expression. PMID- 3034721 TI - [Plasmids and mobile genetic elements of pathogenic Yersinia bacteria]. AB - The review of literature is devoted to molecular and genetic organization of movable genetic elements responsible for synthesis of pathogenicity factors of Yersinia bacteria. General characterization of Yersinia pathogenicity factors is given. We analysed the role of plasmids in expression of the pathogenic properties of these bacteria and the role of IS elements of Yersinia in expression of genes encoding synthesis of the pathogenicity factors. Replicative properties of Yersinia plasmids are described. The role of chromosomal genes in regulation of synthesis of the plasmid-specific pathogenicity factors is discussed. PMID- 3034722 TI - [Transposition of mobile dispersed genes (MDG) after substitution of various chromosome pairs in inbred Drosophila melanogaster strains]. AB - Transpositions of MDG-1, MDG-3 and copia were detected as a result of crosses of the inbred maladaptive LA stock with laboratory stocks, in order to construct the genomes carrying different combinations of the LA or non-La chromosomal pairs. Changes of the mobile gene distributions were revealed in chromosomes of hybrid genotypes, as compared to parental chromosomal pairs. A trivial source of variability of chromosomal molecular structure ensured by crossing over was excluded by inversions which serve as suppressors of crossing over in corresponding crosses. Multiple transpositions of mobile genes in definite chromosomal sites were detected in genotypes carrying chromosomal pair 2 originated from the LA stock. No such transpositions were observed, when the pair 2 was substituted by the chromosome 2 originated from the Swedish-b line or in control crosses, where the LA stock was not involved. Both LA chromosomes 2 and 3 were shown to be the targets of transpositions. Comparison of hot spot transposition sites of MDG-1, as a result of crosses, with the earlier described rare events of spontaneous transpositions in the LA stock, coupled with its fitness increase, revealed that the hot spot sites were shared in both series of experiments. The data obtained show that transpositions of mobile genetic elements may change the genetic and molecular structure of the chromosome involved in crosses, in spite of suppression of crossing over by inversions usually suggested as a tool for keeping chromosomal genetic structure intact. PMID- 3034723 TI - [Insertion mutagenesis in Bacillus subtilis (Marmur)]. AB - A method for obtaining mutations in Bacillus subtilis using integration into the chromosome of the plasmid pHV60 ligated to chromosomal DNA fragments was developed. Auxotrophic mutants acquire, in addition, chloramphenicol-resistance, due to insertion of appropriate plasmid determinants. Chromosomal localization was established and the properties of several mutants were studied. PMID- 3034724 TI - [Reversal of malignant transformation induced by the oncogenic virus SV40. I. Induction of reversal to normal type for the contact inhibition trait]. AB - The possibility of induction by the oncogenic DNA-containing virus SV40 of reversions to normal phenotype as regards contact inhibition ("flat" revertants), was studied in spontaneously transformed chinese hamster fibroblasts. Negative selection was used for detection of revertants. The method adopted allowed to study the mutagenic activity of the virus, while excluding its transforming effect. In all experiments the frequency of revertants after infection exceeded that in control series. The value of induction varied from 1.2 to 28.4 X 10(-6). The tumor promoter 12-O-tetradecanoyl-phorbol-13-acetate (TPA) known to increase the frequency of mutations induced by carcinogens in vitro, displayed no enhancing effect on the frequency of revertants induced by SV40. The lack of enhancement of virus-induced reversions after TPA treatment might be explained by the lack of the transforming effect of SV40 in the system studied. Some of the normal "flat" colonies were T-antigen positive, i. e. the viral oncogene was expressed. The role of mutations induced by SV40 in cellular genes controlling malignancy is discussed. PMID- 3034725 TI - [Comparative study of the transformation of various thymidine kinase-deficient human and animal cell lines with the thymidine kinase gene of the Herpes simplex virus]. AB - The comparative study of transformation of four thymidine kinase deficient cell lines (mouse mammary carcinoma cell line FS tk-; rat cell line Rat-2tk-; mouse cell line Ltk-, clone D1; human cell line 143tk-) with the thymidine kinase cloned gene of Herpes simplex virus 1 was undertaken. The differences in efficiency and optimal conditions of transformation were shown for these cell lines. The advantages and disadvantages of the cell systems examined for the use in experiments for transformation and cotransformation of cultured cells with isolated genes are discussed. PMID- 3034726 TI - Evolution of a mouse Y chromosomal sequence flanked by highly repetitive elements. AB - Mammalian primary sex is determined by the presence or absence of the Y chromosome. However, little is known about the molecular processes through which the Y chromosome exerts its action. We applied recombinant DNA techniques to isolate mouse Y chromosomal fragments and described previously a clone designated as AC11 (Y. Nishioka and E. Lamothe. 1986. Genetics, 113:417-432). To obtain information on DNA sequences that flank AC11, we screened a mouse genomic library for the presence of AC11-related sequences and isolated over 50 positive clones. In this report we describe clones ACC2 and ACC3, both of which contain highly repetitive elements. Using a male-specific portion of these clones, we compared DNA's isolated from mice (Mus musculus, M. hortulanus, M. spretus, M. cookii, M. pahari, and M. platythrix), rat, hamster, and guinea pig and obtained results that agree with the phylogenetic relationships deduced from morphological and biochemical studies. The male-specific accumulation of the related sequences was found only in M. musculus, M. hortulanus, and M. spretus. PMID- 3034727 TI - Transposition studies of mini-Mu plasmids constructed from the chemically synthesized ends of bacteriophage Mu. AB - We describe below the chemical synthesis of the right and left ends of bacteriophage Mu and characterize the activity of these synthetic ends in mini-Mu transposition. Mini-Mu plasmids were constructed which carry the synthetic Mu ends together with the Mu A and B genes under control of the bacteriophage lambda pL promoter. Derepression of pL leads to a high frequency of mini-Mu transposition (5.6 X 10(-2) which is dependent on the presence of the Mu ends and the Mu A and B proteins. Five deletion mutants in the Mu ends were tested in the mini-Mu transposition system and their effects on transposition are described. PMID- 3034728 TI - A new tetracycline-resistance determinant, class E, isolated from Enterobacteriaceae. AB - A fifth tetracycline(Tc)-resistance determinant, designated class E, has been identified on a transferable plasmid found in a fecal strain of Escherichia coli. This determinant does not show homology by DNA-DNA hybridization at high stringency with any of four other Tc resistance determinants (classes A, B, C and D) previously described among the Enterobacteriaceae. Resistance is inducible by 1 microgram Tc/ml and increases the minimum inhibitory concentration 130-fold for Tc and 3.5-fold for minocycline. The mechanism, like that of the other four determinants examined, appears to involve an active efflux of the drug. Using a 32P-labeled cloned fragment containing the resistance determinant, we have found the determinant in Aeromonas, but not in any of over 200 other E. coli strains tested. PMID- 3034729 TI - Fixation of mutations in the viral genome during an outbreak of foot-and-mouth disease: heterogeneity and rate variations. AB - Rates of fixation of mutations during the evolution of the foot-and-mouth disease virus (FMDV) C1 in nature have been estimated by hybridization of viral RNA to cloned cDNAs representing defined FMDV genome segments, and comparison of the selected RNAs by T1 RNase oligonucleotide fingerprinting. Values ranged from less than 0.04 X 10(-2) to 4.5 X 10(-2) substitutions per nucleotide per year (s/nt/yr), depending on the time period and the genomic segment considered. Rates for viral structural protein genes were up to sixfold higher than for nonstructural protein genes. Values in excess of 10(-2) s/nt/yr have been measured for the RNA region that encodes VP1-VP3. The nucleotide sequences of the major immunogenic region of capsid protein VP1 have been determined for six new FMDV C1 isolates, and they are compared with the two previously known sequences of FMDV C1 (C-S8 and C1-O). Both oligonucleotide fingerprinting of selected RNA fragments and direct nucleotide sequencing demonstrate that genetic heterogeneity exists among three viruses isolated on the same day, introducing a significant indetermination in the evaluation of fixation rates of mutations. During the FMDV C1 outbreak, amino acid substitutions did occur that are known to affect the immunological properties of the virus. The proportion of mutations between two viral RNAs does not increase significantly with the time elapsed between the two isolations, suggesting a cocirculation of multiple, related, nonidentical FMDVs ('evolving quasispecies') as the mode of evolution of this agent. PMID- 3034730 TI - Organization and expression of the transforming region from the European elk papillomavirus (EEPV). AB - The nucleotide sequence of the early (transforming) region from the European elk papillomavirus (EEPV) double-stranded DNA has been determined together with flanking regions. The established sequence, which is 5732 bp long, shows that the genome of EEPV is closely related to the previously sequenced bovine papillomavirus type 1 (BPV-1) and deer papillomavirus (DPV) genomes. Seven open reading frames (ORFs), designated E1-E7, were identified in similar positions as in the BPV-1 genome. The E1 and E5 regions were best conserved. The strong homology between the E5 ORFs of EEPV, BPV-1 and DPV is interesting in the light of the recent proposal that these ORFs encode a major transforming function (Schiller et al., 1986; DiMaio et al., 1986). A set of mRNAs, comprising six size classes, were identified in EEPV-transformed cells. At least two different promoters appear to control EEPV transcription in transformed cells. PMID- 3034732 TI - Complete nucleotide sequence and mRNA-mapping of the large subunit gene of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) from Chlamydomonas moewusii. AB - Nucleotide (nt) sequence of the large subunit (LS) gene of ribulose-1,5 bisphosphate carboxylase/oxygenase from the green alga, Chlamydomonas moewusii, and mapping of transcription ends was achieved by two new strategies. The deduced LS sequence of 475 amino acid residues was compared with similar genes from six other species; cyanobacteria, land plants and a related alga (C. reinhardtii). The most conserved regions are the three ribulose bisphosphate binding sites and the CO2 activator site. The nt sequence conservation outside the coding region is limited to only three segments within the 5'-flanking region: a region of tandem repeats, TATAA box and ribosome-binding site. Termination point of transcription is an 'A' residue 3' to the first of two 18-nt inverted repeats, which has the potential to form a stem-loop hairpin structure. The possible role of these potential regulatory features for transcription and translation, and similar structures in other LS genes is presented. PMID- 3034731 TI - Construction of an infectious pseudorabies virus recombinant expressing a glycoprotein gIII-beta-galactosidase fusion protein. AB - An infectious herpesvirus mutant has been constructed in which a major structural envelope glycoprotein gene was replaced by a hybrid gene encoding a novel fusion protein consisting of the N-terminus of the viral glycoprotein joined to Escherichia coli beta-galactosidase (beta Gal). Specifically, we fused DNA encoding the first 157 amino acids of the structural glycoprotein gIII from pseudorabies virus strain Becker to the E. coli lacZ gene in a bacterial expression vector. The resulting hybrid gene was then used to replace the wild type gIII gene in the virus by cotransfection of plasmid and viral DNA. The desired viral recombinants were identified by their inability to react with specific monoclonal antibodies that recognized only wild-type gIII protein. One such mutant virus, PRV-Z1, was chosen for further analysis. PRV-Z1 expressed a glycosylated gIII-beta Gal fusion protein after infection of PK15 cells. The fusion protein has no demonstrable beta Gal activity and, although glycosylated, remains sensitive to the enzyme endo-beta-N-acetylglucosaminidase H, unlike the mature gIII gene product, indicating that the fusion protein was incompletely processed. PMID- 3034734 TI - Isolation of low-copy-number sequences that neighbor satellite DNA in mammals. AB - To investigate the role of satellite DNA in eukaryotic genomes, we isolated from an African green monkey (Cercopithecus aethiops) genomic library cloned segments containing the previously described deca-satellite linked to low-copy-number genomic sequences. Three such clones were obtained. The low-copy-number sequences in the three clones do not cross-hybridize suggesting that they derive from different genomic loci. The structure of one of the clones, lambda MkA, is described in detail. Subcloned segments containing the low-copy-number sequences from lambda MkA anneal to monkey, human and mouse genomic DNA. The subcloned probes were used to select clones containing homologous sequences from a second, independent monkey library as well as from human and mouse genomic libraries. Several of the newly isolated monkey clones hybridized to probes containing the species-specific deca- and alpha-satellites, confirming the genomic association of the low-copy-number sequence in lambda MkA with satellite DNA. Moreover, several of the human and mouse clones hybridized to species-specific human and mouse satellite DNAs, respectively. These experiments indicate that the low-copy number sequence in lambda MkA and its association with satellite DNA is conserved in primates and rodents. PMID- 3034733 TI - Patterns of integration of exogenous DNA sequences transfected into mammalian cells of primate and rodent origin. AB - We studied the cotransfer and cointegration of several genes transfected into four cell lines of primate origin. Mouse thymidine-kinase-negative LM cells, which had been extensively studied previously, were used as a reference. We found that in monkey kidney Vero cells, on average between 3.5 and 6.0 kb of plasmid sequences was integrated per clone, while in the murine LM cell line, 9-186 kb of exogenous DNA was integrated per clone. Transformed Vero clones which had integrated more than 6 kb of DNA did not integrate larger DNA fragments in a second transformation assay than had the parental Vero cells. We found that the efficiency of gene cointegration is similar in Vero, HeLa and GM4312A cells, the latter being deficient in the repair of UV-induced damage. The human hepatocarcinoma Hep G2 cells integrated on the average 2 kb more exogenous DNA than the three other primate cell lines, which resulted in a 4-5 times higher efficiency of gene cointegration. Plasmid penetration and persistence in a free state between 24 h and two weeks after transfection was similar in Vero and LM cells. No major post-integration DNA rearrangement could be demonstrated after the isolation of Vero clones. These observations correlate the low efficiency of gene cointegration in some primate cell lines with a genomic recombination step or with rearrangements taking place during early cell divisions following integration. PMID- 3034735 TI - Plasmid vector pBR322 and its special-purpose derivatives--a review. AB - The plasmid pBR322 was one of the first EK2 multipurpose cloning vectors to be designed and constructed (ten years ago) for the efficient cloning and selection of recombinant DNA molecules in Escherichia coli. This 4363-bp DNA molecule has been extensively used as a cloning vehicle because of its simplicity and the availability of its nucleotide sequence. The widespread use of pBR322 has prompted numerous studies into its molecular structure and function. These studies revealed two features that detract from the plasmid's effectiveness as a cloning vector: plasmid instability in the absence of selection and, the lack of a direct selection scheme for recombinant DNA molecules. Several vectors based on pBR322 have been constructed to overcome these limitations and to extend the vector's versatility to accommodate special cloning purposes. The objective of this review is to provide a survey of these derivative vectors and to summarize information currently available on pBR322. PMID- 3034736 TI - Expression of human gastrin gene in normal and gastrinoma tissues. AB - The complete sequence of the human gastrin gene is reported here. This gene consists of three exons. Nine Alu family sequences are found within the gene and in the surrounding region. S1 mapping study showed that the transcription of gastrin gene starts at 60, 57, or 55 bp upstream from the 3' end of the first exon. The mechanism of mRNA synthesis in a gastrinoma tissue was studied to clarify the ectopic production of gastrin. It was found that mRNA synthesis starts from the same three transcriptional start points. Southern blotting profiles for normal gastric antrum and gastrinoma DNA were indistinguishable from each other within the 18-kb region containing the gastrin gene, showing that no genomic rearrangements are associated with the gastrinoma formation. Thus, the overproduction of gastrin in this tumor is likely to be due to an aberrant expression control system of the cell, rather than a change in the control region of the gastrin gene. PMID- 3034737 TI - A novel multistep method for generating precise unidirectional deletions using BspMI, a class-IIS restriction enzyme. AB - A novel approach is described that permits the introduction of unidirectional deletions into a cloned DNA fragment, in a precisely controlled manner. The method is based on the use of a special vector and a class-IIS restriction endonuclease, BspMI, which produces staggered cuts 4 and 8 nucleotides (nt) to the 3' from its recognition site 5'-ACCTGC-3'. The DNA fragment is inserted into the pUC19-based plasmid, which contains a unique BspMI recognition site, and the appropriate number of cleavage-and-deletion cycles is performed, each cycle removing 4 bp. Since the recognition site is not affected by the BspMI cleavage, no recloning of the DNA fragment is necessary. Each cycle consists of BspMI cleavage, removal of the 4-nt single-stranded cohesive ends with mung bean nuclease (MB), and blunt-end ligation to recircularize the plasmid. The shortened plasmid is reintroduced into the host, after one or after several such 4-bp deletion cycles. When DNA is inserted into the multiple cloning site in the lacZ alpha gene, the progress of 4-bp removal can be followed by determining the Lac phenotype, since removal of multiples of 3 bp retains the reading frame while other kinds of deletions distort (or restore) the reading frame. Loss of pre existing restriction sites or creation of new ones also permits monitoring the progress of the deletion process.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3034738 TI - The production of generalized transducing phage by bacteriophage lambda. AB - Generalized transduction has for about 30 years been a major tool in the genetic manipulation of bacterial chromosomes. However, throughout that time little progress has been made in understanding how generalized transducing particles are produced. The experiments presented in this paper use phage lambda to assess some of the factors that affect that process. The results of those experiments indicate: the production of generalized transducing particles by bacteriophage lambda is inhibited by the phage lambda exonuclease (Exo). Also inhibited by lambda Exo is the production of lambda docR particles, a class of particles whose packaging is initiated in bacterial DNA and terminated at the normal phage packaging site, cos. In contrast, the production of lambda docL particles, a class of particles whose packaging is initiated at cos and terminated in bacterial DNA, is unaffected by lambda Exo; lambda-generalized transducing particles are not detected in induced lysis-defective (S-) lambda lysogens until about 60-90 min after prophage induction. Since wild-type lambda would normally lyse cells by 60 min, the production of lambda-generalized transducing particles depends on the phage being lysis-defective; if transducing lysates are prepared by phage infection then the frequency of generalized transduction for different bacterial markers varies over a 10-20-fold range. In contrast, if transducing lysates are prepared by the induction of a lambda lysogen containing an excision defective prophage, then the variation in transduction frequency is much greater, and markers adjacent to, and on both sides of, the prophage are transduced with much higher frequencies than are other markers; if the prophage is replication defective then the increased transduction of prophage-proximal markers is eliminated; measurements of total DNA in induced lysogens indicate that part of the increase in transduction frequency following prophage induction can be accounted for by an increase in the amount of prophage-proximal bacterial DNA in the cell. Measurements of DNA in transducing particles indicate that the rest of the increase is probably due to the preferential packaging of the prophage proximal bacterial DNA. These results are most easily interpreted in terms of a model for the initiation of bacterial DNA packaging by lambda, in which the proteins involved (Ter) do not recognize any particular sequence in bacterial DNA but rather recognize some feature of the DNA tht is sensitive to lambda exonuclease, such as a nick or a double-stranded cut.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3034739 TI - ACE-inhibitors in the treatment of elderly hypertensives. AB - Angiotensin-converting enzyme inhibitors are gaining acceptance as safe and effective agents for treatment of hypertension. Data on their use specifically in elderly hypertensives, however, are limited. Addressed in this review of the available literature are the questions whether they are effective hypotensives in the elderly, whether their effects are age-related, and what effects, if any, do they have on morbidity and mortality in geriatric hypertension. PMID- 3034740 TI - Aging is without effect on the pituitary-adrenal axis in men. AB - The effects of ovine corticotropin releasing factor (o-CRF) on the secretions of ACTH, cortisol, dehydroepiandrosterone (DHEA) and dehydroepiandrosterone sulfate (DHEA-S) were studied in the healthy young and elderly male individuals. o-CRF caused significant elevations of plasma ACTH, cortisol and DHEA concentrations, in all subjects. Moreover, there were no significant differences in the time dependent responses of plasma ACTH, cortisol and DHEA to o-CRF between the young and elderly male individuals. o-CRF showed no significant rise of plasma DHEA-S, in all subjects. We conclude that the pituitary-adrenal sensitivity to the hypothalamic CRF in the elderly does not change with aging. PMID- 3034742 TI - [Neurologic manifestations of cervical osteochondrosis in the clinical picture of vibration disease]. PMID- 3034741 TI - Peritoneal reaction to liquid silicone: an experimental study. AB - Silicone oils (polymethylsiloxanes) with different viscosities (200, 1000 and 5000 cSt) and a fluorinated silicone oil (5000 cSt) were injected into the peritoneum of CBA mice. Three weeks after the injection, peritoneal exudates were obtained and investigated using light and electron microscopy. For each silicone oil type, light-microscopic examination revealed the presence of abundant extracellular lipid-like droplets and a dense cellular infiltrate composed of macrophages, lymphocytes and multinucleated giant cells. Lipid-like inclusions were observed in macrophages and multinucleated giant cells. These intracellular inclusions were revealed to be membrane bound by transmission electron microscopy. We observed no differences with respect to the amount of phagocytotic activity for the different silicone-oil types tested. Purified silicone oils of lower viscosity (OP 200 and OP 1000) resulted in the mildest chronic inflammatory reaction. Our results suggest that emulsification, phagocytosis and granulomatous inflammation are associated with intraocular silicone-oil implantation. The question of which properties would make a silicone oil most suitable for clinical use requires further investigation. PMID- 3034743 TI - [Pathological changes in the cervix uteri in human papillomavirus infection (HPV)]. PMID- 3034746 TI - Pelvic angiography in malignant trophoblastic disease. AB - Pelvic angiography was analyzed in 24 patients with malignant trophoblastic disease. Abnormal findings such as prominent uterine arteries, hypervascularity of the uterus, arteriovenous shunts, tumor staining and pooling were observed. Translucency was observed in all patients with choriocarcinoma. This finding was observed in only 3 of 11 patients (27.3%) with invasive mole and was not apparent in patients with an undetermined group. These differences were statistically significant. Translucency of pelvic angiography in malignant trophoblastic disease was more suggestive of choriocarcinoma than of invasive mole. Thus, translucency in pelvic angiography may be a distinctive finding with which to differentiate choriocarcinoma from invasive mole, without resection of the uterus. PMID- 3034744 TI - Effect of oxytetracycline administration on intestinal metabolism of oestrogens and on plasma sex hormones in healthy men. AB - The effect of oxytetracycline (1 g/day for five days) on the enterohepatic recycling of oestrogens and on plasma sex hormone concentrations was assessed in healthy men. Plasma oestrone (E1), oestradiol-17 beta (E2), 4-androstenedione (A), 5 alpha-dihydrotestosterone (5 alpha-DHT), total and free testosterone (T and free T), binding capacity of sex hormone binding globulin, luteinizing hormone, dehydroepiandrosterone-sulphate, urinary total E1, E2, and oestriol (E3), and oestriol-3-glucuronide (E3-3G) and faecal unconjugated and conjugated E1, E2, and E3 were measured by radioimmunoassay (RIA). Treatment with the antibiotic significantly increased the excretion of faecal conjugated oestrogens, which parallelled a decrease in urinary oestrogen excretion, especially of E3. The effect on urinary E3 could be explained almost entirely by the simultaneous decrease of urinary E3-3G concentrations. In urine and faeces the E2/E3 and E1 + E2/E3 ratios increased, probably because of the diminished reductive metabolism of oestrogens in the gut. No significant effects on plasma unconjugated oestrogen concentrations were observed. Moreover, in the present study oxytetracycline had no remarkable effect on plasma total, or free T concentrations, nor on other plasma hormones measured. Our results suggest that enterohepatic recycling and intestinal metabolism of oestrogens may be significant in men. The mechanism of action of antibiotics on oestrogen metabolism probably involves decreased hydrolysis by beta-glucuronidase of oestrogen conjugates by the intestinal contents, diminishing the reabsorption of aglycones of oestrogen conjugates and resulting in faecal loss of the steroids. PMID- 3034745 TI - Cytomegalovirus colitis and oesophageal ulceration in the context of AIDS: clinical manifestations and preliminary report of treatment with Foscarnet (phosphonoformate). AB - Three patients with biopsy diagnosed invasive cytomegalovirus infection of the colon have been seen in the context of the acquired immune deficiency syndrome (AIDS). Cytomegalovirus colitis presented with fever, abdominal distention, bloody diarrhoea and weight loss. Plain abdominal radiographs showed generalised large bowel dilatation in one patient. Cytomegalovirus infection was shown histologically, but the virus could not be cultured from the stool; no other gastrointestinal pathogens could be demonstrated. The patients were treated with a 14 day continuous infusion of Foscarnet 0.08 mg/kg/min (phosphonoformate, Astra Pharmaceuticals). One patient showed a partial response to therapy, but the cytomegalovirus colitis relapsed; the second patient had a symptomatic response only and the third patient died of non-cytomegalovirus opportunist infection while on treatment. Two other patients with biopsy proven cytomegalovirus ulceration of the oesophagus were seen, presenting with dysphagia, fever and weight loss. Invasive infection of the gastrointestinal tract with cytomegalovirus is now a major clinical problem in AIDS. Treatment with Foscarnet may be initially effective, but does not eliminate cytomegalovirus infection. PMID- 3034747 TI - Characterization of the nuclear progesterone receptor in normal human endometrium after progesterone injection. AB - Nuclear progesterone receptors (RPN) were characterized in the 0.4 M KCl nuclear extract of the endometrium using [3H]R5020 as a ligand, in women in the proliferative phase of the cycle (n = 5), in the midcycle (n = 5), in the secretory phase (n = 4), and in premarin-treated women (n = 3), 1-3 h following progesterone injection. The receptor character of the binding was demonstrated by high specificity for progestin binding, high saturable affinity for R5020 and a sedimentation constant of about 3S in sucrose density gradient. The RPN characteristics were similar in all 4 groups and were also similar to those of the remaining cytosolic RP. Despite the fact that total RP levels differed in all 4 groups, the percentage of the receptor that was measured in the nuclear extract was similar in all groups and ranged from 52 to 61% of the total RP levels. PMID- 3034748 TI - Opioid control of luteinizing hormone secretion in patients with hirsutism and hyperandrogenemia. AB - The present study was designed to evaluate the influence of hyperandrogenemia on the activity of the opioid system regulating LH secretion in menstruating women. Ten subjects presenting with hirsutism and hyperandrogenemia and 9 healthy normally cyclic subjects participated in the study. Naloxone or saline was administered on 2 different days both during the follicular (6-8 days after menstrual bleeding) and during the luteal phase of the menstrual cycle. Naloxone significantly increased plasma LH levels in the luteal, but not during the follicular phase of the cycle in both subject groups. It may be inferred from these observations that opioid-mediated inhibition of LH secretion is not altered in menstruating hyperandrogenic patients, suggesting that the circulating androgens are not an important determinant of the functional neuroendocrine activity of the opioid system. PMID- 3034750 TI - A retrospective study of drugged driving in Norway. AB - The National Institute of Forensic Toxicology, Oslo, receives blood and urine samples from all Norwegian drivers apprehended on suspicion of driving under the influence of alcohol or drugs. In 1983 we received samples from 1446 drug suspected drivers, out of which 445 underwent toxicological analysis. The drugs found most frequently were tetrahydrocannabinol (THC) (n = 199), diazepam (n = 166) and amphetamine (n = 102). A cautious interpretation of the data indicate that about 200 of the 445 subjects selected for toxicological analysis drove under severe influence of drugs. Because of the high percentage of submitted cases not analysed for drugs, this figure represents a minimum estimate. Compared with the results from 1978, we found a several-fold increase in detections of THC and amphetamine in 1983. The number of diazepam detections did not increase in a similar way, but we estimated that the diazepam detections would have increased 3 fold if we had analysed as frequent for this drug in 1983 as in 1978. PMID- 3034751 TI - [Virus infections caused by transfusion of blood and blood products. 2: Parvovirus, cytomegalovirus, Epstein-Barr virus, human immunodeficiency virus]. PMID- 3034749 TI - [Behavioral pharmacological action of Ca-4-(3,5-dihydroxy-3-methylpenthylamido) butyrate (mevalonic GABA, MV-GABA)]. AB - The behavioral pharmacological effects of Ca-4-(3,5-dihydroxy-3 methylpenthylamido) butyrate (mevalonic-GABA, MV-GABA), a new GABA derivative were studied in comparison with those of Ca-hopantenate (HOPA) in mice. MV-GABA had no effect on the general behavior and electric shock-induced fighting behavior. The dosage of MV-GABA which caused locomotor hypoactivity produced an impairment of the rotarod performance. MV-GABA inhibited the hyperactivity induced by a dopamine (DA) agonist (methamphetamine) and acetylcholine (ACh) antagonists (scopolamine and atropine) at a dose which did not affect locomotor activity in normal mice. MV-GABA prolonged the pentobarbital-Na-induced sleeping time, and it prolonged the latencies until convulsion and death after administration of strychnine. MV-GABA and HOPA antagonized the electroconvulsive shock-induced amnesia in the passive avoidance response of mice. These results suggest that MV-GABA has effects on the central nervous systems, in particular, ACh and DA neural systems. The actions of MV-GABA were qualitatively similar to those of HOPA except for the effect on the DA neural system. PMID- 3034752 TI - [Studies on accessory function of B cells]. PMID- 3034753 TI - Beta cell response to the hyperglycaemic clamp in three patients with insulinoma: a study using a hyperglycaemic glucose clamp. AB - To determine the mechanism responsible for deficient carbohydrate metabolism in patients with insulinoma, we studied three affected patients and seven normal controls using the hyperglycaemic clamp method (8.4 mmol/l) with the BIOSTATOR (GCIIS). In insulinoma patients, the amount of glucose necessary to reach the hyperglycaemic clamp was less than that required in normal controls (6.19 +/- 1.19 mg/min/kg vs. 9.95 +/- 0.53 mg/min/kg) (p less than 0.05). There was no significant difference in metabolized glucose (M) in the stable phase of the hyperglycaemic clamp; however, the M/IRI in this phase was less in those with insulinoma (7.9 +/- 0.50) than in controls (22.26 +/- 4.14) (p less than 0.05). There was no difference in beta cell secretory response to hyperglycaemic stimulus (defined as the increase in the concentration of C-peptide from the basal state to the stable phase of the hyperglycaemic clamp) between the two groups. Hepatic insulin extraction was significantly lower in patients with insulinoma than in normal controls (+0.72 +/- 0.07 vs. +0.85 +/- 0.01). Finally, the ratios of fractional turnover of glucose (K/IRI); glucose clearance/IRI and total rate of elimination of glucose from the extracellular pool/IRI were also all lower in patients with insulinoma than in controls (p less than 0.05). These data support the conclusion that deficient glucose metabolism seen in these patients is not related to a lack of response to glucose on the part of normal or neoplastic islet tissue.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3034754 TI - The regulation of adipose tissue pyruvate dehydrogenase activity of dietary fiber. AB - In vitro studies have established that insulin enhances the oxidation of pyruvate to acetyl CoA by the stimulation of mitochondrial pyruvate dehydrogenase (PDH) activity through plasma membrane binding response (Jarett and Seals 1979; Kiechle, Jarett, Dennis and Kotagal 1980). In the present study adipose tissue PDH activity was utilized as a marker for insulin responsiveness. The metabolic response of this enzyme to exogenous insulin was employed to test the hypothesis that dietary fiber enhances tissue responsiveness to insulin using adipose tissue from male weanling Sprague Dawley rats. Eight groups of rats (n = 5 per group) were fed ad libitum various diets containing different levels of cellulose and protein as already reported elsewhere (Ogunwole, Knight, Adkins, Thomaskutty and Pointer 1985). Percent insulin stimulation of PDH from basal activity (PDS) was utilized as an index of insulin responsiveness. Compared to all fiber treated groups, both basal (PDB) and insulin stimulated (PDI) activities were significantly lower (P less than 0.05) in the fiber free groups at both low (10%) and high (20%) protein levels. At all fiber levels tested (0, 5, 15 and 30%) protein intake resulted in a significant increase in both PDB and PDI. Gradual increase in cellulose intake resulted in a biphasic increase in PDS in both protein groups at the 5% and 30% fiber levels. PDS was higher (P less than 0.05) in the 10% protein groups than the 20% protein group at all fiber levels tested. A significant interaction effect of protein and fiber was observed on PDB (P less than 0.001) and PDI (P less than 0.04) when caloric intake was held constant as a covariate.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3034755 TI - Plasma beta-endorphin in response to oral glucose tolerance test in obese patients. AB - In order to clarify the possible interaction between endogenous opioids and glucose homeostasis in obesity we studied Beta-Endorphin (B-Ep), ACTH, cortisol and insulin plasma levels in response to an oral glucose tolerance test (OGTT) in 8 females suffering from uncomplicated obesity and in 6 healthy volunteers of normal weight. Results were evaluated in terms of secretion areas subtracted from basal value. Basal glucose, insulin and B-Ep levels were significantly higher in the obese patients compared to controls, cortisol levels and ACTH were not statistically different between obese and normal subjects. During OGTT total areas of insulin secretion were significantly higher in the obese patients; cortisol, ACTH, B-Ep plasma levels did not change in controls, whereas obese patients showed a response to B-Ep which reached a peak at 60 minutes. The area of B-Ep response to OGTT in obese patients was significantly higher than in controls. On the basis of these results we may suggest that the opioid system belongs to the chain of neuroendocrine and metabolic events responsible for the origin and the growth of overweight. But the possibility exists that obesity itself can enhance the B-Ep secretion above all through overeating. In this regard it is to stress that glucose ingestion induces in obese patients, differently from normal subjects, insulin hypersecretion and the B-Ep secretion, possibly from gastro-enteric tract and/or pancreatic isles. PMID- 3034756 TI - Chronobiological analysis of changes in circadian rhythm of anterior pituitary hormones level during glycemia normalization by means of biostator. AB - We compared the circadian rhythms of anterior pituitary hormones in 15 patients with noncompensated insulin-dependent diabetes on first and second day treatment with Biostator. The rhythm was evaluated by means of a least squares analysis and presented as the circle of cosinors. In noncompensated diabetes the TSH and prolactin rhythm was maintained, whereas other hormones of the anterior pituitary showed no significant rhythm. In the course of one-day normalization of glycemia by means of Biostator the TSH and prolactin rhythm was maintained, whereas the circadian rhythm of growth hormone and ACTH levels appeared with acrophase at 18.47 and 19.59 hour, respectively. The LH rhythm did not exist, whereas the FSH rhythm was dubious. One may assume that noncompensated diabetes results in the impairment of certain pituitary hormonal rhythms and these disturbances are reversible after restoring of normoglycemia. PMID- 3034758 TI - Corticotropin-releasing factor in humans. II. CRF stimulation in patients with diseases of the hypothalamo-pituitary-adrenal axis. AB - A stimulation test with 100 micrograms ovine or human corticotropin-releasing factor (CRF) is a useful diagnostic tool in diseases of the hypothalamo-pituitary adrenal axis. No serious side effects were observed during the test procedure. The results showed that the CRF test is useful in making the differential diagnosis of established Cushing's syndrome (n = 42). The CRF test was also repeated after transsphenoidal surgery in 25 patients with Cushing's disease. Successfully operated patients exhibit no, blunted or normal adrenocorticotropic hormone (ACTH) responses to CRF (n = 15), whereas patients who did not show remission remained hyperresponsive (n = 10). In patients with autonomous adrenal cortisol secretion, the ACTH response to CRF was suppressed (n = 10). After surgery the ACTH response to CRF can already be demonstrated when cortisol levels are still undetectable. Pulsatile administration of CRF in one patient after unilateral adrenalectomy and another patient under corticoid therapy revealed that ACTH responses to CRF normalize rapidly but cannot be sustained if CRF administration is withdrawn, suggesting that the cause of adrenal failure after unilateral adrenalectomy for Cushing's syndrome or long-term corticoid therapy is due to hypothalamic CRF deficiency. The decrease of the ACTH responses to CRF in glucocorticoid-treated patients correlated directly to the daily corticoid dosage. Since the ACTH hyperresponse to CRF in 6 patients with Cushing's disease was also suppressed by short-term dexamethasone treatment, the pituitary level as target site for the acute feedback inhibition is also demonstrated. The evaluation of the CRF-induced ACTH response in patients with secondary adrenal failure without detectable pathology in the sella and suprasellar region (n = 6) enables the differentiation between hypothalamic and pituitary adrenal insufficiency. In patients with hypothalamic lesions the ACTH response to CRF was normal whereas insulin hypoglycemia failed to induce an ACTH rise. PMID- 3034757 TI - Enhanced response of plasma prolactin to metoclopramide during chronic converting enzyme inhibition. AB - The effect of chronic converting enzyme inhibition with enalapril on the PRA, PRL and plasma aldosterone responses to metoclopramide was studied in 10 patients with mild to moderate essential hypertension. Enalapril reduced supine blood pressure and increased heart rate significantly. PRA and urinary sodium excretion rose significantly. PRA levels did not change after metoclopramide neither during placebo nor during enalapril. The aldosterone response to metoclopramide was not altered by enalapril, indicating that this response is independent of the renin angiotensin system. The PRL response to metoclopramide was considerably enhanced after 4 weeks of treatment with enalapril. It is proposed that enalapril, by decreasing the formation of angiotensin II, increases the prolactin reserve. PMID- 3034759 TI - Asymptomatic shedding and subsequent transmission of genital herpes simplex virus. AB - We report the transmission of genital herpes simplex virus (HSV) infection from an asymptomatic woman shedding virus from the cervix to two male sexual partners and further transmission from these two men while their infection was in the prodromal phase. The value of the restriction enzyme analysis of viral deoxyribonucleic acid (DNA) is presented. Guidelines regarding the management of patients who are found to be asymptomatic shedders of HSV are discussed. PMID- 3034760 TI - Bowenoid papulosis: clinical and histological study of eight cases. AB - Eight cases of bowenoid papulosis are reported. The clinical diagnoses were confirmed by histology. In one case an immunoperoxidase method showed the presence of papillomavirus antigen in the nucleus of the most superficial epidermal cells. We consider bowenoid papulosis to be a condition with specific features that distinguish it clinically and histologically from carcinoma in situ and condylomata acuminata. PMID- 3034761 TI - Sexually communicable micro-organisms in human semen samples to be used for artificial insemination by donor. AB - Two hundred and thirty seven semen samples from 10 institutes for artificial insemination by donor (AID) in Belgium and the Netherlands were tested for the presence of Neisseria gonorrhoeae, Chlamydia trachomatis, Mycoplasma hominis, Ureaplasma urealyticum, herpes simplex virus, and cytomegalovirus. The incidence of these micro-organisms in the semen samples was 0%, 6.3%, 4.6%, 35.9%, 0%, and 0.4% respectively, and 47% of all samples were infected with one or more of the micro-organisms. As the ejaculates from which the samples had been taken had already been, or would be, used for AID, the exclusion of microbiological contamination with sexually communicable micro-organisms before insemination is indicated. PMID- 3034762 TI - Clinical and microbiological evaluation of 46 episodes of genital ulceration. AB - In 44 consecutive patients 46 episodes of genital ulceration were studied. The presumptive clinical diagnosis was evaluated by extensive microbiological investigations. In 15 (33%) episodes the clinical diagnosis did not accord with the microbiological one. Chancroidal lesions were most commonly found to have other microbiological aetiology. Secondary invasion of treponemal or viral lesions by other bacterial species (mostly anaerobes or pyogenic cocci) or genital pyodermia were the cause of confusion in six of nine cases of chancroid. In the other three a ducreyilike bacterium was found. Direct Gram staining of ulcer material did not help the diagnosis of chancroid. The implications of the results of this study for clinical practice are discussed. PMID- 3034763 TI - Polybrominated biphenyls. PMID- 3034764 TI - Mevalonate phosphorylation in lemon grass leaves. PMID- 3034765 TI - Induction of choline-ethanolamine kinase in chicken liver by 17-beta-estradiol. PMID- 3034766 TI - Induction of secondary immune response by reactivated Japanese encephalitis virus in latently infected mice. AB - Development of secondary immune response has been studied following reactivation of latent Japanese encephalitis virus (JEV) infection in mice. The virus could be reactivated in 43% of the latently infected mice at 27 weeks p.i. by treatment with cyclophosphamide. The reactivated virus induced delayed-type hypersensitivity (DTH) and leucocyte migration inhibition (LMI) responses in mice, with peak activity on Day 5 post-reactivation (p.r.). The DTH persisted at low levels for long periods. Humoral immunity measured by haemagglutination inhibiting antibody showed a four-fold rise in antibody titres. DTH was transferable by immune spleen cells for 5 days p.r. only. It is, therefore, concluded that JEV reactivation generates a quick and short-lived secondary immune response. PMID- 3034767 TI - Catabolism of homologous murine monoclonal hybridoma IgG antibodies in mice. AB - Two neutralizing monoclonal hybridoma antibodies to the surface spike glycoprotein E2 of the JHM strain of murine hepatitis virus were injected into homologous BALB/c mice, and their biological half-lives were determined by sequential titration of plasma samples in a virus-specific enzyme immunoassay. Intravascular half-lives of monomeric immunoglobulins were estimated at 8.0 +/- 1.5 days for antibody 5B19.3, an IgG1, and 12.7 +/- 2.4 days for antibody 4B11.6, an IgG2a. These catabolic rates are statistically different from each other (P less than 0.001) and significantly higher than previously reported values, which were all obtained with radiolabelled polyclonal or myeloma immunoglobulins of unknown specificities. Failure to remove aggregated 4B11.6 antibodies by high speed centrifugation yielded a statistically significant acceleration of biological turnover (half-life 9.9 +/- 1.6 days; P less than 0.01). PMID- 3034768 TI - Interferon-alpha enhances the production of leukotriene B4 in murine peritoneal macrophages stimulated by opsonized zymosan. AB - Murine peritoneal macrophages pretreated with interferon (IFN)-alpha and then stimulated by opsonized zymosan produced two to three times more LTB4 than untreated macrophages. However, PGE2 production was not changed by IFN-alpha. Meanwhile, IFN-gamma did not affect the production of LTB4 and PGE2. From the results it is considered that IFN-alpha can modulate inflammation or host defence through the production of LTB4. PMID- 3034770 TI - Human adult T-cell leukaemia virus (ATLV) genome codes for the protein pX27 structurally homologous to immunoglobulin-like part of HLA-DQ antigen. AB - Within the 3' end of nucleotide sequence of human adult T-cell leukaemia virus (ATLV) four small open frames in various phases were found previously. The frames code for 10,000-, 11,000-, 12,000- and 27,000-dalton polypeptides, one of which can be responsible for virus transforming activity. By means of methods of prediction of secondary structure from amino acid composition and sequence, the beta-pleated structure was shown to be the main conformation of the 27,000-dalton polypeptide (pX27 ATLV). As a result of fitting of beta-pleats and cysteines in the N- and C-terminal parts of pX27 ATLV to those of C gamma 2 and C gamma 3 domains of human immunoglobulins G (HuIgG) and of immunoglobulin-like domains of human major histocompatibility antigens (HLA-B, -DR, -DQ), a statistically valid homology was observed by comparing the C-terminal part of pX27 ATLV with the immunoglobulin-like domain of HLA-DQw1 antigen. PMID- 3034769 TI - Immune response to uncoupled peptides of foot-and-mouth disease virus. AB - Uncoupled synthetic peptide representing the sequence of amino acids 141-160 of foot-and-mouth disease virus (FMDV) protein VP1 induced a virus-neutralizing antibody response in guinea-pigs. This response required incomplete Freund's adjuvant (IFA) for the primary inoculation and was dependent on the presence of an added cysteine residue with an unblocked sulphydryl group at the carboxy terminus. Secondary immunization could be carried out in the absence of adjuvant. A study of the relative activities of nested sets of uncoupled peptides from 150 160 to 135-160 and 141-160 to 141-155 indicated that amino acids 146-156 were critical for the induction of virus-neutralizing antibodies and that extension to 137-160 further improved this response. Results of in vitro proliferation studies demonstrated that the carboxy-terminal residues on this peptide may form a T-cell epitope. The significance of these observations in the broader context of synthetic peptide vaccines is discussed. PMID- 3034772 TI - Inhibition of superoxide anion production from activated phagocytes by antiinflammatory drugs. PMID- 3034773 TI - Effects of phagocytized substances on insulin secretion in mice. PMID- 3034771 TI - Interplay between extracellular -SH groups and Ca2+ in their mitogenic action on murine lymphocytes: the role of SiO2. AB - Murine spleen lymphocytes, cultured under serum-free conditions and stimulated with concanavalin A (Con A), showed an increased demand for supplemental concentrations of Ca2+ when 2-mercaptoethanol (2-ME) or L-cysteine were also present in the culture medium. The requirement for additional Ca2+ ions was particularly evident at high cell concentrations. The addition of SiO2 (Aerosil 200), in concentrations depending on the original level of Ca2+ in the culture medium, could substitute for the increased demand on calcium ions. This dependency of SiO2 effects on calcium levels in the medium was also evident when supraoptimal, i.e. inhibitory concentrations of silica were tested. Results of experiments with 45Ca suggest that silica particles may act as intercellular carriers of calcium. In contrast to spleen lymphocytes, Con A-stimulated thymus cells exhibited neither a need for an increase in the concentration of Ca2+ nor a dependency on the presence of 2-ME in the medium. PMID- 3034774 TI - Hepatoblastoma in an adult female. PMID- 3034775 TI - Computer aided radionuclide angiography for diagnosis of thyroid function. PMID- 3034776 TI - Specificity of secretory immunoglobulins in cervical mucus of women with uterine cervical dysplasia. PMID- 3034777 TI - Central alpha 2-adrenergic stimulation increases neurointermediate lobe immunoreactive beta-endorphin in spontaneously hypertensive rats. AB - A possible influence of the central alpha 2-adrenergic system on beta-endorphin was examined in rat anterior pituitary, neurointermediate lobe, and plasma. The concentration of beta-endorphin in anterior pituitary, neurointermediate lobe, and plasma was determined by radioimmunoassay 15 minutes after subcutaneous injection of clonidine in 14-week-old spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY). Clonidine reduced the concentration of the plasma beta endorphinlike immunoreactivity in SHR and to a lesser extent in WKY. No significant changes in the concentration of beta-endorphinlike immunoreactivity were observed in anterior pituitary. Clonidine increased the concentration of neurointermediate lobe beta-endorphinlike immunoreactivity in SHR in a dose related manner but did not affect the concentration in WKY. Administration of yohimbine (1 mg/kg) completely blocked the clonidine-induced increase of neurointermediate lobe beta-endorphinlike immunoreactivity in SHR, while prazosin (1 mg/kg) had no effect. These data suggest that the central alpha 2-adrenergic activation increases the neurointermediate lobe concentration of beta endorphinlike immunoreactivity in SHR by suppressing beta-endorphin release from the neurointermediate lobe into the circulation. PMID- 3034778 TI - Inhibition of renin release by analogues of adenosine in rabbit renal cortical slices. AB - Renal cortical slices obtained from male New Zealand rabbits were used to investigate the role of adenosine in the regulation of renin release. Isoproterenol produced a significant (p less than 0.01), twofold to threefold increase in renin release, that was both dose-dependent and time-dependent. Addition of either the l-phenylisopropyl or the N6-ethylcarboxamido derivative of adenosine attenuated this stimulation at concentrations as low as 10(-9) M or 10( 8) M, respectively. Higher doses of d-phenylisopropyladenosine (10(-6) M) or adenosine (10(-5) M) were necessary to significantly reduce the beta-adrenergic response (p less than 0.01). Inhibition was absent in slices preincubated with 10(-5) M 8-phenyltheophylline, a concentration that had no effect on either basal or stimulated renin release. The site of inhibition appeared to be distal to beta adrenergic and prostaglandin receptors since l-phenylisopropyladenosine (10(-8) M) blocked stimulation by selective beta-adrenergic receptor agonists, prenalterol (10(-6) M) or salbutamol (10(-5) M), and by prostaglandin E1. These data suggest that adenosine and its analogues inhibit renin release and that this inhibition may be mediated by a receptor-dependent action on a common point in the pathway leading to release. PMID- 3034779 TI - Influence of neutrophil cationic proteins on generation of superoxide by human polymorphonuclear cells during phagocytosis. AB - The cationic proteins from neutrophil lysosomes have been shown to modulate phagocytic activity of granulocytes. The present study reports the effects of the cationic protein fractions on the generation of O2- by human PMNs during phagocytosis. Human PMNs were reacted with different phagocytic stimuli in the presence and absence of lysosomal cationic proteins and the amount of O2- generated was determined by superoxide dismutase inhibitable reduction of cytochrome c. Total cationic protein extract from neutrophil lysosomes enhanced O2- generated by PMNs during the phagocytosis of IgG-coated latex beads and opsonized zymosan particles. The analysis of the fractions of cationic proteins obtained from a Sephadex G-75 column showed that the O2- generation-enhancing activity was associated with the proteins eluted in fractions III and IV. A protein fraction mainly eluted in void volume inhibited the cytochrome c reduction by O2- formed during phagocytosis. This was due to the presence of superoxide dismutase-like activity since O2- generated by the xanthine-xanthine oxidase system was also inhibited by this fraction. The cationic protein fractions III and IV from the Sephadex G-75 column were further subfractionated. Although the O2(-)-enhancing activity was eluted in the same fractions as chymotrypsin activity, there was no quantitative correlation between the amount of O2- generation and chymotrypsin activity. Moreover, commercial chymotrypsin did not enhance O2- generation. Electrophoretic analysis of the isolated protein fractions suggests that O2- generation enhancing protein (SGEP) is different from lysozyme or chymotrypsin and probably represents previously undescribed protein. PMID- 3034781 TI - Enhanced oxidative burst without interleukin 1 production by normal human polymorphonuclear leukocytes primed with muramyl dipeptides. AB - Purified polymorphonuclear (PMN) cells were obtained from human blood leukocytes by centrifugation on colloidal silica gradients. PMNs could be primed for PMA triggered oxidative burst by muramyl peptide molecules (MDP) and two of its adjuvant active nonpyrogenic derivatives. The priming effect of MDP could be demonstrated after a 1-h incubation period, whereas monocytes needed an 18-h incubation to produce an enhanced response in the NBT reduction test. Only the monocyte-enriched population was able to produce IL-1 activity after muramyl peptide stimulation. Under such conditions, PMNs neither produced nor secreted IL 1-like activity, and no IL-1 inhibitor was present in the supernatant fluids. In conclusion, muramyl peptides were able to prime PMNs for oxidative burst but not to stimulate IL-1-like factor production. PMID- 3034780 TI - Generation of superoxide by immunologically stimulated normal human neutrophils and possible modulation by intracellular and extracellular SOD and rheumatoid factors. AB - Rheumatoid synovial fluids generated significantly greater amounts of superoxide, lysosomal enzymes, and superoxide dismutase from neutrophils into extracellular fluid than osteoarthritic synovial fluids. Rheumatoid factors isolated from serum suppressed superoxide-generating activity of performed immune complexes, but did not suppress that of intermediate-sized immune complexes isolated from RA serum. Synovial fluid neutrophils has a greater capacity to generate superoxide and lower intracellular superoxide dismutase activity, compared with peripheral neutrophils of the corresponding patients. These results suggest that neutrophil superoxide release may be modulated, both by rheumatoid factor and by intracellular and extracellular superoxide dismutase. PMID- 3034782 TI - Luminol-independent chemiluminescence by phagocytes is markedly enhanced by dexamethasone, not by other glucocorticosteroids. AB - The effect of several glucocorticosteroids on the generation of reactive oxygen species (ROS) was examined. The ROS assessed were O-2, H2O2, OH., and chemiluminescence (CL) (determined in the presence or absence of luminol), generated by both opsonized zymosan-stimulated neutrophils or monocytes and by the xanthine-xanthine oxidase system. Except for luminol-independent CL, only high concentrations (10(-4) M) of steroids could decrease each ROS. In contrast, luminol-independent CL generation in the phagocyte system was increased in a dose dependent manner by the addition of dexamethasone, but not by any other steroid. Further, in lymphocyte cultures stimulated with Con A for four days, luminol independent CL generation was demonstrated and enhanced by the addition of dexamethasone, although CL generation was not detected in the absence of dexamethasone. These findings provide evidence that CL does not always represent light specific to ROS, and they suggest the possibility that dexamethasone induces emission of light at sites of inflammation. PMID- 3034783 TI - Studies of phagocytosis in chronic granulomatous disease. AB - Abnormal phagocyte function in chronic granulomatous disease (CGD) is associated with decreased bactericidal activity. Ingestion of serum-opsonized organisms is reported to be normal in these patients. We previously showed that in CGD the expression of C3b receptors (CR1) on polymorphonuclear leukocytes (PMNs) is significantly depressed. In this study, we compared the phagocytic activity of the PMNs from normal healthy controls with that of CGD patients and one individual with myeloperoxidase (MPO) deficiency. The ingestion of sheep erythrocytes (E) by PMNs adherent to a glass surface was examined; the E were coated either with excess IgG (E-IgG) or with C3b plus limited IgG (EAC3b-IgG). The PMNs, both in CGD and in MPO deficiency, ingested E-IgG and EAC3b-IgG at levels markedly above normal. C3b-coated erythrocytes were not phagocytosed. Preincubating the PMNs with sodium azide, which blocks MPO, or catalase, a scavenger of H2O2, caused a marked increase in phagocytosis by normal PMNs. Azide had a variable effect on PMN activity in CGD and no effect on the activity in the subject with MPO deficiency. Even in the presence of azide, the ingestion of EAC3b-IgG by the PMNs from the CGD patients was significantly greater than that seen in paired normals [mean phagocytic index (PI), 2.13 for CGD vs. 1.48 for normals; P less than 0.05 by the paired sample t test]. Similar results were obtained with ingestion of E-IgG. Notably, ingestion of serum-opsonized Candida organisms (relatively nondegradable particles) was markedly above normal with CGD PMNs and, in normal PMNs, azide treatment also evoked an increase. In addition, rosette formation of the adhered PMNs with E-IgG was enhanced with CGD and the azide-treated normal PMNs. We demonstrated that this increased activity was not the result of increased Fc receptor (FcR) number, as determined from the binding of a monoclonal anti-FcR antibody. Both the E-IgG rosette formation and the ingestion by CGD PMNs were abrogated in the presence of an H2O2-generating system. In contrast, the phagocytic activity of MPO-deficient PMNs was not altered by exogenous H2O2. These findings suggest that cellular products generated by the H2O2-MPO-halide system down-regulate the rosette-forming and phagocytic activity of PMNs from normal healthy individuals, but not that from CGD and MPO-deficient patients. PMID- 3034784 TI - Dynamics of chemotactic peptide-induced superoxide generation by human monocytes. AB - Addition of the chemotactic peptide, f-Met-Leu-Phe, to human monocytes induced a burst of superoxide release, which ceased after approximately 3 min. Diminished responsiveness to f-Met-Leu-Phe, but not to phorbol myristate acetate (PMA), was induced by 1- to 3-h storage at 0 degrees C or by 2 min in 40 microM adenosine (ADO). Reversal of the ADO block was achieved by addition of adenosine deaminase (ADA) as little as 15 sec before the f-Met-Leu-Phe stimulus; ADA had no effect when added poststimulus. The ADO experiments suggest that there are a minimum of two sequentially produced intermediates in the f-Met-Leu-Phe stimulus-response pathway. The first intermediate persists for less than 30 sec. The second, formation of which is stimulated by the first, persists for the duration of the response and is the target of ADO inhibition. The ADO target is apparently not protein kinase-C, since the response of inhibited cells to PMA was unimpaired. The maximal inhibition by adenosine of f-Met-Leu-Phe-induced superoxide generation was approximately 50%. It is possible that f-Met-Leu-Phe stimulates two pathways of NADPH activation, only one of which is inhibited by adenosine. PMID- 3034786 TI - Carcinoma in situ and cancer of the testis. Workshop. Copenhagen, August 1986. PMID- 3034787 TI - Cell lines of human germinal cancer. AB - Three new established human germ cell tumour lines, H 12.1, H 12.5 and H 12.7, are described. Cytogenetic and growth characteristics, morphology, histology, and tumour marker and steroid hormone production in vitro and/or in vivo revealed properties commonly found in other germ cell tumour lines and in patients with these tumours. In vitro oestrone and oestradiol were produced by the H 12 lines and four other lines (833 KE, 1156 Q, 1428 A and 2102 EP) under high and low density (differentiation inducing) conditions. AFP was produced by one line and beta-hCG by six of seven lines under low density conditions. The pattern of oestrogen production suggests that these hormones could be useful in AFP and beta hCG negative patients. Differentiated elements of somatic and extraembryonic character, observed in tumours of H 12.1 and H 12.7, underline the value of these lines in the study of differentiation and other germ cell tumour related questions. PMID- 3034785 TI - Isolation of two immunogenically different allergens from Schistosoma japonicum eggs. AB - Two allergenic components, termed J1 and J2, were isolated from a soluble egg antigen preparation (SEA) of Schistosoma japonicum by anion-exchange chromatography on DE52 and gel chromatography on Sephacryl S-200. The apparent molecular weights of J1 and J2 were 260,000 and 46,000, respectively, by gel chromatography on Sephadex G-150. By SDS-polyacrylamide gel electrophoresis, both J1 and J2 showed apparent homogenicity and their estimated molecular weights were 135,000 and 45,000, respectively. The isoelectric point of J1 (pI 4.9) was similar to that of J2 (pI 4.8). Both J1 and J2 bound to Con A-Sepharose 4B, indicating their glycoprotein nature. The amino acid compositions of J1 and J2 have some similarities. However, phenylalanine and leucine, which contain large hydrophobic groups, were dominant in J1, whereas serine and threonine, which contain a hydroxyl group, were dominant in J2. J1 was sensitive to heating or pronase treatment, whereas J2 was rather stable to these treatments. Both J1 and J2 were sensitive to 0.1 M periodate treatment. When mice were immunized with either J1 or J2 with A1(OH)3 as an adjuvant, anti-J1 or anti-J2 IgE antibody was highly specific to the respective antigens. Since S. japonicum-infected mouse serum has high PCA titer to J1 and J2, these two components are the major allergens of S. japonicum eggs. PMID- 3034788 TI - Flow cytometry derived DNA content of the primary lesions of advanced germ cell tumours. AB - We currently lack good prognostic indicators for patients with advanced disseminated germ cell tumours (GCT). An analysis of archival tumour samples of forty-nine patients with advanced GCT for flow cytometry derived DNA content suggests an inverse relationship between tumour proliferative activity and patient survival. The great majority of patients with advanced GCT have aneuploid tumour subpopulations. This analysis suggests that the survival of patients with advanced GCT may depend in part on kinetic variables present in the primary tumour. PMID- 3034789 TI - Proliferation of human testicular tumours. AB - Whole tumour perfusion with radioactive DNA precursors reveals that proliferation of human testicular tumours differs widely depending on type and stage of development. In seminomas a homogeneous distribution of proliferating cells at early stages is followed by a shift of the growth fraction to the periphery at advanced stage. Embryonal carcinomas show a chessboard-like pattern of proliferating fractions with high 3H-thymidine labelling indices and short tpot. In mature teratomas proliferation is low. It increases in immature teratocarcinomas. Lack of proliferation is observed in beta-hCG-positive syncytiotrophoblastic cells of choriocarcinomas, the growth fraction being limited to the cytotrophoblastic compartment. PMID- 3034790 TI - Comparative antitumour activity of cisplatin and two new cisplatin-analogues JM8 and JM9 in human testicular carcinoma xenografts. AB - The comparative antitumour activity of cisplatin, JM8 and JM9 was tested using a panel of different heterotransplanted human testicular tumour cell lines. All drugs were applied at equitoxic doses in a 5 day schedule. In the two cisplatin sensitive cell lines 2102 EP and H 12.1 both analogues were inferior to cisplatin. No significant therapeutic effect was achieved with any of the three drugs in the cisplatin resistant line H 23.1. Thus JM8 and JM9 seem to be less active in cisplatin sensitive tumours and seem to be of no advantage in the case of cisplatin resistance. PMID- 3034791 TI - Carcinoma-in-situ of the testis: possible origin from gonocytes and precursor of all types of germ cell tumours except spermatocytoma. AB - Based on evidence from morphological and histochemical studies and from clinical experience, the following hypotheses are proposed: carcinoma-in-situ (CIS) germ cells are malignant gonocytes; these CIS gonocytes have some capacity to regress into more primitive, totipotent embryonic cells which can give rise to all types of nonseminomatous germ cell tumours; the tumour germ cells of classical seminomas are malignant gonocytes derived from CIS gonocytes which have lost their ability to regress into totipotent embryonic cells; the ability of CIS gonocytes to regress into totipotent embryonic cells decreases with age, whereas the capacity to form classical seminoma cells is preserved; the transformation of CIS gonocytes into invasive tumours is dependent on factors such as gonadotrophins and/or testicular steroids; the pathogenesis of classical and spermatocytic seminoma are unrelated. As a consequence of these hypotheses an alternative nomenclature for carcinoma-in-situ, seminoma and dysgerminoma is suggested. PMID- 3034792 TI - Incidence of bilateral testicular germ cell cancer in Denmark, 1960-84: preliminary findings. AB - The incidence of a second primary testicular germ cell cancer in the contralateral testicle among 2338 men with a first primary testicular germ cell cancer diagnosed in the years 1960-79 in Denmark was established in this preliminary report. The material represents 83% of the total cohort followed until 31 December 1984. The relative risk for a patient with testicular cancer to get yet another testicular cancer was studied, taking into account the histology of the first primary testicular germ cell cancer. Based on fifty-eight nonsimultaneous contralateral testicular cancer cases and 19,995 'person-years at risk', the overall relative risk of invasive germ cell cancer in the contralateral testicle following a first germ cell testicular cancer was found to be 23.3 (95% confidence interval: 18-30). Among men with nonseminoma the risk was higher (relative risk = 27.5) than among men with seminomas (relative risk = 20.1). Overall, sixty-two (2.7%) patients developed a second cancer. In four of these patients bilateral tumours occurred simultaneously. PMID- 3034793 TI - Multivariate analysis of prognostic variables in patients with disseminated non seminomatous testicular cancer: results from an EORTC multi-institutional phase III study. PMID- 3034796 TI - Retroperitoneal lymph node dissection in clinical stage IIA and IIB nonseminomatous germ cell tumours of the testis. AB - From 1980 to 1984 inclusive, ninety-one consecutive evaluable patients underwent primary retroperitoneal lymphadenectomy for clinical stage IIA or IIB nonseminomatous germinal testicular cancer. Nodes were negative in twenty cases (22%), and forty-seven patients (52%) were treated with chemotherapy either postoperatively (thirty clinically understaged patients) or at relapse (seventeen cases). After a median follow-up period of nearly 5 years (range 18-78 months) the disease-free survival was 98%. None of thirty patients with radiographic abnormalities greater than or equal to 3 cm in the retroperitoneal nodes had negative histology, and twenty-two (73%) were treated with chemotherapy. Preoperative serum levels of AFP and hCG were not useful in selecting patients with positive nodes. Primary chemotherapy is now used in patients with radiographic evidence of retroperitoneal metastases greater than or equal to 3 cm. PMID- 3034795 TI - Orchidectomy alone versus orchidectomy plus radiotherapy in stage I nonseminomatous testicular cancer: a randomized study by the Danish Testicular Carcinoma Study Group. AB - All Danish patients with stage I nonseminomatous testicular cancer diagnosed between December 1980 and January 1984 entered a randomized study comparing irradiation of retroperitoneal lymph nodes with surveillance only after orchidectomy. Twenty-four of the seventy-nine patients in the observation-only group have relapsed, three patients relapsing more than 2 years after orchidectomy. Ten of the seventy-three patients receiving irradiation have relapsed, all within 10 months after orchidectomy. The median time to relapse in both groups was 4.5 months. Irradiation prevented retroperitoneal relapses. Thirty-three of the relapsed patients were rendered disease free with chemotherapy, and one is still being treated. Four deaths have occurred, all unrelated to testicular cancer or antineoplastic treatment. Absence of embryonal carcinoma and presence of teratocarcinoma correlated with improved relapse-free survival. Patients with increased serum concentrations of tumour markers before orchidectomy had an increased risk of relapse. Surveillance-only is a reasonable treatment strategy in clinical stage I nonseminomatous testicular cancer. Preferably control and treatment of relapses should take place in specialized centres. PMID- 3034797 TI - An analysis of poor risk assignment in patients with germ cell tumours. AB - Four methods for assigning patients with germ cell tumours to poor risk programmes were compared using 118 consecutive patients treated on 'good' and 'poor' risk studies. The median survival of patients deemed poor risk by Memorial Sloan-Kettering Cancer Center was 11 months, by Indiana University 15 months, by the National Cancer Institute 15 months, and by the European Organization for Research on the Treatment of Cancer 23.5 months. The major differences between these criteria were in the use of pretreatment tumour marker values, and the use of specific sites of metastases as independent prognostic variables. These data imply that a variable number of good prognosis patients enter some poor risk trials, the number being dependent upon the criteria used. Poor risk criteria should be as restrictive as possible in order to avoid over treating good risk patients, and to avoid artificially inflated response rates resulting from the successful treatment of such poor risk patients. PMID- 3034794 TI - Surveillance following orchidectomy for stage I testicular cancer. AB - Surveillance following orchidectomy was introduced in the management of Stage I testicular nonseminoma in 1979 and Stage I seminoma in 1983. Of 132 nonseminoma patients followed for 12-84 months (median 43 months) the relapse rate is 27%. Relapses were diagnosed 2-44 months after orchidectomy with 90% of relapses appearing within the first year. Of the 132 patients, 131 are alive and disease free. The pattern of relapse was as follows: 47% of relapses occurred in abdominal nodes, 13% in abdominal nodes and lung, 17% in the lung and 23% with elevated serum markers as the only evidence of disease; 26% of relapsing patients had normal serum AFP and hCG levels. The prognostic significance of thirteen clinical histopathological and biochemical factors has been analysed by multiple regression analysis. Histology and lymphatic invasion within the primary tumour are significant independent prognostic factors. A total of thirty-six patients had scrotal interference prior to removal of the primary tumour. This was not a contra-indication to surveillance. None has developed scrotal recurrence and the overall relapse rate (11%) is comparable to that observed in the surveillance series as a whole. Fifty-two patients with Stage I seminoma have been observed from 12-41 months after orchidectomy. Seven (13%) have relapsed and six of the seven relapses have been confined to retroperitoneal lymph nodes. Preliminary data suggests that pre-orchidectomy elevation of serum hCG is not a significant prognostic factor. PMID- 3034798 TI - The function of the primordial germ cell in extragonadal tissues. AB - The identity of the germ cell tumours of the pineal and the thymus with those of the testis and ovary suggests that the widely disseminated primordial germ cells might subserve some special function in these sanctuaries. It is proposed that thymic localization might be required for the conveyance of genetic haematological and immunological information and that the pineal-diencephalic localization could programme neuroophthalmic tissues prior to the development of the blood-brain barrier. The latter speculation was tested by producing allergic encephalomyelitis in thymectomized, bursectomized and thymobursectomized chickens. It was found that although thymectomy and, to a lesser extent, bursectomy decreased the severity of the experimental encephalomyelitis the combined procedure resulted in more severe inflammatory lesions. This may be due to release of suppressed intraneural immunological mechanisms in the somatically impaired bird. PMID- 3034799 TI - Intermediate filament proteins as tissue specific markers in normal and neoplastic testicular tissue. AB - Normal testicular tissue and primary and metastatic testicular germ cell tumours were examined for their intermediate filament protein (IFP) expression. Seminomas were shown to react with antibodies to vimentin, while non-seminomatous germ cell tumours were strongly positive with antibodies to cytokeratin. In the case of teratocarcinoma, several components of the tumour can be distinguished using a combination of monoclonal and polyclonal antisera in the double-label immunofluorescence technique. We conclude that antibodies to cytokeratin and vimentin can be helpful in the diagnosis of testicular germ cell tumours, especially in the differentiation between seminomas and non-seminomatous testis tumours. PMID- 3034800 TI - Cytogenetic studies of human testicular germ cell tumours. AB - In a search for consistent cytogenetic alterations in testicular germ cell cancers we have thus far studied some twenty surgical specimens of seminomas and nonseminomatous tumours. From the literature and our results it is now clear that such testicular tumours generally have a hyperdiploid to hypotriploid chromosomal content, and frequently possess a possibly site-specific chromosomal marker, an isochromosome 12p. A significant correlation between the presence of the i(12p) and advanced clinical stages has been revealed in our study. Several other chromosomal regions are consistently involved in cytogenetic changes: 1p and 1q, 6q, 7p, 9q, 12p, 17q, and 22q. Although there is little doubt that characteristic chromosomal lesions exist in testicular germ cell tumours, the impact which specific lesions may have on tumour progression is still unclear. PMID- 3034802 TI - HLA phenotype and clinicopathological behaviour of germ cell tumours: possible evidence for clonal evolution from seminomas to nonseminomas. AB - Analysis of 101 patients with germ cell tumours of the testis who have been typed for HLA DR antigens has provided confirmatory evidence for an association of DR5 with the development of seminoma and demonstrated an association of DR7 with metastases. These observations taken with the suggestion of HLA linkage in the small numbers of familial cases reviewed, does suggest that there is an HLA linked gene involved in the clinicopathological behaviour of germ cell tumours. Though of only theoretical interest at present these observations may be of considerable importance in the future given the observation in mice that transfection of missing MHC genes into a malignant tumour can produce a vaccine that enables previously unexposed animals to resist the original malignant tumour (Hui et al., 1984). PMID- 3034801 TI - Expression of growth regulatory genes in primary human testicular neoplasms. AB - Seven testicular tumours of different histological type--two seminomas, two teratomas/teratocarcinomas/embryonal carcinomas, one mixed seminoma/teratoma, one Leydig cell tumour and one testicular lymphoma--were examined for the expression of four potentially growth regulatory genes by Northern blotting. Seven out of seven testicular tumours contained transcripts that hybridized with a human insulin cDNA-probe whereas only four out of seven tumours contained IGF II transcripts. One tumour contained high levels of LDL-receptor transcript whereas all seven tumours contained significant quantities of HMG-CoA-reductase mRNA. PMID- 3034804 TI - Distribution pattern of human papilloma virus 16 genome in cervical neoplasia by molecular in situ hybridization of tissue sections. AB - Using a highly sensitive method with single-stranded RNA probes, we analyzed the distribution pattern of HPV 16 DNA by in situ hybridization in CIN II (10 cases), CIN III (11 cases) and in invasive cervical carcinoma (17 cases). The technique used detected as little as 20-50 viral genomes per cell. This sensitive technique unmasked HPV 16 genomes in the basal cells of all forms of CIN. In CIN III viral genomes were present throughout the entire thickness of the epithelium. There was a striking difference in the distribution of viral DNA in CIN II compared with CIN III and invasive cancer. Variable viral genome distribution was observed in CIN II with the highest copy number in the area of epithelial differentiation. In contrast, CIN III showed a uniform distribution pattern of HPV genomes reflecting the lack of epithelial maturation. The majority of invasive carcinomas showed the same uniform distribution of the HPV 16 genomes as CIN III. PMID- 3034803 TI - Presence of in vivo-activated T-cells expressing HLA-DR molecules and IL-2 receptors in peripheral blood of patients with nasopharyngeal carcinoma. AB - Epstein-Barr Virus (EBV) is associated with two malignant diseases, African Burkitt's Lymphoma (BL) and Undifferentiated Nasopharyngeal Carcinoma (UNPC). North Africa is a geographical area with a high incidence of NPC. Our purpose in this study was to explore cell-mediated immunity of peripheral blood lymphocytes (PBL) from patients with UNPC and DNPC. We found an elevated percentage of OKT8 cells and of large granular lymphocytes (LGL) (30-35% HNK-I-positive cells) compared to PBL from healthy matched individuals. PBL from NPC patients contained 35% HLA-DR-positive and 30% Interleukin-2 (IL-2) receptor-positive circulating lymphocytes. PBL from NPC patients exhibited a normal proliferative response to phytohemagglutinin (PHA) and Concanavalin A (Con A) and an increased response to pokeweed mitogen (PWM). Natural killer (NK) activity towards K562 cells was low in our patients who, in addition, exhibited no lytic activity against HLA-matched EBV-transformed B cells. This lack of cytotoxicity against an EBV-transformed B cell line cannot be explained by an impairment of IL-2 secretion, and is probably a result of the presence of high numbers of OKT8 suppressor T cells. PMID- 3034806 TI - Tumor-cell-induced platelet aggregation is a glycoprotein-dependent and lipoxygenase-associated process. AB - To characterize the platelet receptor sites and the platelet metabolic pathways involved in tumor-cell-induced platelet aggregation, we have used a homologous system consisting of human platelets and 2 tumor cell lines of human origin, which activate platelets through different mechanisms. Preincubation of platelets with an MAb against platelet glycoprotein Ib partially blocked tumor-cell-induced platelet aggregation, and preincubation of platelets with an MAb against the glycoprotein complex GPIIb/IIIa totally blocked the aggregation induced by the 2 tumor-cell lines. No inhibitory effect was found when platelets were treated with PAF-receptor antagonists or with specific peptides which block the platelet sites involved in bacterially induced platelet aggregation. Compounds which raised intra-platelet cAMP levels inhibited tumor-cell-induced platelet aggregation in a dose-related manner. Inhibition of cyclo-oxygenase by aspirin which blocked TxB2 formation by platelets did not inhibit platelet aggregation induced by tumor cells whereas the BW755 compound which inhibits cyclo- and lipoxygenase blocked platelet aggregation. These results demonstrate that tumor-cell-induced platelet aggregation is a glycoprotein-dependent and a lipoxygenase-associated phenomenon. PMID- 3034805 TI - Immune control of SV40-induced tumors in mice. AB - The ability of mice to mount a cytotoxic T-lymphocyte (CTL) immune response to SV40 T-antigen is determined by the H-2 haplotype of the host; H-2b and k mice are high responders and H-2d mice are low responders. Mice of these 3 H-2 haplotypes were challenged with SV40 and their ability to generate and sustain an antibody response to SV40 T-antigen was found to be equivalent. To investigate the role of the different components of the host immune response in controlling growth of SV40-induced tumors, the tumorigenic potential of freshly established cell lines, obtained by SV40 transformation of cells from normal tissues of inbred strains of mice of 6 H-2 haplotypes, was assessed. Each cell line was tumorigenic in athymic and newborn mice but not in adult syngeneic immunocompetent mice. Cells from these initial SV40-transformed lines were then passaged in athymic (nu/nu) mice, re-established in vitro and again transferred into syngeneic animals. Transfer of H-2d SV40 transformants to low or non responder mice of the H-2d haplotype resulted in tumor formation in some animals. Cells derived from these tumors expressed both the viral encoded T-antigen and the H-2Dd restriction element. Furthermore, the proportion of animals with tumors varied with the strength of their CTL-responsiveness to SV40 T-antigen in association with H-2Dd. Therefore, in H-2d animals, tumor cell growth appears to result from escape of cells from inefficient CTL surveillance. No tumors were formed by transfer of the in vivo selected H-2b or H-2k SV40 transformants to syngeneic high-responder mice. We therefore investigated the role of CTL in the selection of SV40-transformed cells able to escape immune surveillance. Under conditions of stringent immune selection by CTLs, tumorigenic cells that no longer expressed the relevant H-2 class-I restriction element were obtained. Although interaction between the various immune effector mechanisms may play a role in the recognition and elimination of SV40 transformants, our results were consistent with the hypothesis that the SV40-specific CTL response is the predominant control of SV40 tumor growth. PMID- 3034808 TI - The relationship between serum enalaprilat concentration and the hypotensive effect in man. AB - The relationship between serum concentrations of the angiotensin converting enzyme inhibitor, enalaprilat, and its antihypertensive action was investigated using integrated pharmacokinetic-dynamic modelling techniques. The model was parameterized in terms of slope "m", describing the sensitivity to the fall of blood pressure, and an intercept term "i". A linear pharmacodynamic model: E = m. ce(t) + i, adequately characterized the concentration-effect relationship in most of the young (aged 22-30 years) and elderly normotensive subjects (aged 65-73 years). However, no age-related differences were seen, indicating that angiotensin II vasoconstrictor action is unaltered by senescence. PMID- 3034807 TI - c-yes and bcl-2 genes located on 18q21.3 in a follicular lymphoma cell line carrying a t(14;18) chromosomal translocation. AB - The c-yes-1 and bcl-2 genes have been independently mapped at the same position of the chromosome band, 18q21.3, which is a breakpoint in the t(14;18) chromosomal translocation. High-molecular-weight genomic DNAs and mRNA isolated from a follicular lymphoma cell line FL-18 and an Epstein-Barr virus harboring FL 18EB cell line, which carried the t(14;18), were analyzed. Although there was no detectable c-yes rearrangement, the chromosome 18-specific probe b (bcl-2) revealed one rearranged fragment after EcoRI, BamHI, PstI and SstI digestion. Therefore, breakage of chromosome 18 in the t(14;18) occurred at the SstI fragment containing the probe b sequence on the bcl-2 gene. The transcript of c yes was undetectable, whereas active transcription of bcl-2 was observed in the FL-18 and FL-18EB cell lines at a higher level than in other types of lymphoma cell line. These data suggest that the c-yes-1 is not involved in the t(14;18), and the 2 genes located at 18q21.3 are independent sequences. PMID- 3034809 TI - Imipramine receptors in human platelets: effect of age. AB - Imipramine receptors were studied in platelets from six healthy young subjects (age between 24 and 38 years), five newborns, and six healthy elderly persons (age between 70 and 81 years). Binding parameters, the maximum binding capacity (Bmax) and the apparent dissociation constant (Kd), were determined by Scatchard's analysis. Level of differences between young subjects and the other groups was determined by Student's t-test. Bmax (mean +/- SD) was 1162 +/- 138 (young persons), 564 +/- 65 (newborn), and 508 +/- 98 (elderly persons) fmol/mg protein. The figure for the young was different from that of the newborn (p less than 0.001) and the elderly (p less than 0.01). Kd (means +/- SD) was 1.78 +/- .69 (young persons), 0.68 +/- 0.13 (newborn), and 0.80 +/- 0.27 nM in the elderly. Kd in the volunteers was different from that in the newborn or the elderly subjects (p less than 0.01). Imipramine receptors in platelets appear to be influenced by development and aging. PMID- 3034810 TI - Divergent effects of human lymphokine derived oligopeptides on PMNLs function of young and aged healthy subjects. AB - The effects of low molecular weight lymphokine-derived oligopeptides (LK-OPs) on PMNLs effector functions and receptor mediated biochemical events were studied in the case of healthy young and aged subjects. In the case of young subjects the low mol. wt. LK-OPs stimulated the Fc gamma receptor-mediated effector functions of PMNLs, whereas an inhibition was observed in PMNLs of the elderly (extracellular cytotoxicity and intracellular killing). The underlying biochemical events, induced by low mol. wt. LK-OPs stimulation, were also investigated. In PMNLs of young subjects the oxidative metabolism was stimulated (enhanced O2 consumption, O2 and H2O2 production) by low mol. wt. LK-OPs, while in elderly subjects it was inhibited. The cyclic nucleotides regulating the Fc gamma receptor mediated effector functions and the oxygen radicals formation showed an altered dynamic response under low mol. wt. LK-OPs stimulation with aging i.e. the cGMP level could not be changed at all. Our results suggest that low mol. wt. LK-OPs induced inhibition of Fc gamma receptor mediated effector functions with aging could be partly explained by an altered post receptorial coupling switch and as a consequence the lymphokines could not play their role of immunomodulators further impairing the altered immune response with aging. PMID- 3034811 TI - Methadone vs morphine: comparison of their effect on phagocytic functions. AB - A comparison of the effects of methadone and morphine on phagocytic physiology was carried out in mice, using a number of tests, to estimate the risk of using methadone in maintenance protocols for opiates addicts. Results indicate that methadone, like morphine, reduces (a) R.E.S. activity and (b) PMN superoxide anion production, while unlike morphine it (a) does not produce haematologic changes, (b) does not exacerbate C. albicans infections, (c) does not inhibit phagocytosis and killing by murine polymorphonuclear leukocytes and macrophages, or by rabbit alveolar macrophages, and (d) does not reduce spleen and liver weight. These results are in strict agreement with those previously found in human subjects receiving controlled administration of morphine or methadone. Compared to morphine methadone therefore appears to have a lower toxic potentiality. PMID- 3034812 TI - Inhibition of delayed hypersensitivity reactions by a new agent, cis-1-methyl-4 isohexylcyclohexane carboxylic acid (IG-10)--II. The mechanism regarding the action on lymphokines. AB - A newly synthesized anti-allergic agent, cis-1-methyl-4-isohexylcyclohexane carboxylic acid (IG-10), has the capacity to inhibit the effector phase of delayed hypersensitivity reactions. In the present paper, the effect of IG-10 was studied on the generation of superoxide anion (O2) from macrophages, on macrophage chemotaxis, and on the activity of lymphokines such as skin reactive factor (SRF), macrophage migration inhibitory factor (MIF) and monocyte/macrophage chemotactic factor (MCF) in guinea pigs. Oral administration of IG-10 (50-200 mg/kg) inhibited SRF-induced skin erythema in a dose-dependent manner. In vitro, this agent (10(-7) -10(-5) g/ml) did not inhibit the generation of O2- from macrophages. The agent (10(-7) -10(-6) g/ml) significantly inhibited the activity of MIF and this inhibition was not due to the facilitation of normal migration. For macrophage chemotaxis, IG-10 (10(-8) -10(-6) g/ml) significantly inhibited MCF-induced chemotaxis. The agent also depressed the macrophage chemotaxis induced by N-formyl-methionyl-leucyl-phenylalanine but not the chemotaxis induced by E. coli culture filtrate. The inhibitory action of IG-10 on MCF activity was not influenced by antiglucocorticoid agents such as 17 alpha methyltestosterone and androstenedione which reverse significantly the inhibitory action of glucocorticoids. The inhibitory action of IG-10 was relatively dependent on exogenous Ca2+ and Mg2+, and was antagonized by dbc-GMP. PMID- 3034813 TI - Radiation-induced DNA damage and its repair. AB - Application of modern methods of organic chemistry and recombinant DNA technologies has provided new insights in the field of DNA radiation damage and its repair. An overview of the chemical nature of the lesions inflicted on DNA by ionizing radiation is presented. The structures of 29 different DNA modified base or sugar residues are shown in comprehensive formation schemes. A fraction of radiation-induced modified bases is spontaneously released from the DNA chain during irradiation. Another part remains attached to the DNA chain backbone and for its characterization mild formic acid or enzymatic hydrolysis have been used. Starting from the chemical formulae of the altered base residues, the specific repair enzymes and their modes of action are discussed. Various glycosylases and endonucleases have been purified to homogeneity, and in some cases the gene which encodes the protein cloned. Using methods derived from Maxam and Gilbert sequencing procedures and DNA fragment 32P-labelled at one end, it has been shown that the alkali-labile sites in DNA induced by radiation are strongly dependent on the DNA base sequence. Enzymatic methods have been used to analyse the DNA base defects produced by gamma-irradiation of cells under in vivo conditions. Structures of modified bases were the same as those observed when DNA was irradiated in aqueous solution. PMID- 3034814 TI - The effect of different dietary levels of vitamin A on metabolism of copper iron and zinc in the chick. AB - Chicks were fed on diets containing either no added vitamin A or 3300 micrograms/kg or 330,000 micrograms/kg retinol equivalents for 30 d. Concentrations of copper, iron and zinc were higher in liver and lower in plasma at low and high intakes of vitamin A. Haemoglobin, packed cell volume and erythrocyte levels were depressed by both low and high vitamin A intake and could be related to vitamin A levels by quadratic equations. The Zn and Fe levels in erythrocytes and serum albumin and ceruloplasmin were also affected in a similar fashion by low or high vitamin A diets. Hepatic activity of alcohol dehydrogenase (EC 1.1.1.1) and cytochrome oxidase (EC 1.9.3.1) paralleled Zn and Cu concentrations respectively. Superoxide dismutase (EC 1.15.1.1) and hydrolysis of triolein and retinyl palmitate were not correlated significantly with concentrations of metals but were correlated negatively with log vitamin A concentration. No changes in bone concentrations of Cu, Fe or Zn were detected. It is suggest that vitamin A influences metabolism of Cu, Fe and Zn possibly, in part, due to a decrease in secretion of transport proteins by the liver. PMID- 3034815 TI - Comparative efficacy of handwashing agents against cytomegalovirus. AB - Conscientious handwashing is often recommended as an important method for limiting transmission of cytomegalovirus (CMV) from infected individuals to health, education, and child care professionals. To assess the efficacy of handwashing, fingertips of radiation-sterilized latex gloves were inoculated with 0.2 mL of ten different CMV strains. Virus in each inoculum was quantitated by plaque assay. After five minutes, viral inocula were allowed to remain (control), or were washed away by dropwise application of 10 mL of distilled water (DI), 5 mL of 0.08% soap followed by 5 mL of DI, 5 mL of 0.01% chlorhexidine gluconate followed by 5 mL of DI, or 5 mL of 0.025% povidone-iodine solution followed by 5 mL of DI. Separate glove fingertips were sampled 5, 15, 30, 60, 120 and 240 minutes after washing and cultured in duplicate for CMV. Similar studies were performed using human cadaver skin. Ordinary soap was as effective at preventing CMV recovery as other more expensive agents. For inocula with less than 5 log10 pfu CMV/mL, washing with water alone was as effective as other agents. This was confirmed in similar studies with human hands using five CMV stains. Handwashing is probably an effective method for removing CMV from contaminated hands. PMID- 3034816 TI - Biochemical transmitters regulating the arrest and resumption of meiosis in oocytes. PMID- 3034818 TI - Cell type and proton relaxation. PMID- 3034817 TI - [Drug-induced intrahepatic cholestasis caused by flecainide acetate and enalapril]. PMID- 3034819 TI - Etoposide for small cell lung cancer. PMID- 3034820 TI - Leukocyte migration inhibition detects cross-reacting antigens between cells transformed by Epstein-Barr virus (EBV) and EBV-like simian viruses. AB - The leukocyte migration inhibition (LMI) technique was used to measure the T cell mediated immune response of Epstein-Barr-virus (EBV)-seropositive human donors to antigens associated with B cell lines of simian origin, transformed by simian EBV like viruses, Herpesvirus papio (HVP), H. pan, H. gorilla and H. pongo. Extracts of cell lines carrying three of the four simian viruses (from gorilla, chimpanzee and orangutan) induced a positive LMI response, whereas lines carrying baboon derived HVP were ineffective. None of the simian virus-transformed lines elicited an LMI reaction in human EBV-seronegative individuals. Leukocytes from patients with acute infectious mononucleosis (IM) failed to respond to any of the lines transformed by EBV or the simian EBV-like viruses. Such lines express the virally encoded nuclear antigen, but have only a low level of viral cycle-associated antigens. Extracts of the EBV-carrying human cell line P3HR-1 induced with 12-O tetradecanoylphorbol-13-acetate to express high levels of early and virus capsid antigens (EA, VCA, respectively) however, elicited a strong response with leukocytes from patients with acute IM. During convalescence, IM patients became responsive to EBV, H. gorilla-, H. pan- and H. pongo-transformed lines, indicating that the LMI reaction induced by these simian virus-transformed lines was directed against the antigens expressed in immortalized cells rather than against antigens of the lytic cycle. It is highly probable that this reaction reflects a cross-recognition of the nuclear antigens associated with these four transforming viruses (excluding H. papio) at the level of human T cells. PMID- 3034822 TI - Relationship of Theiler's murine encephalomyelitis viruses to the cardiovirus genus of picornaviruses. AB - Sequence analysis of VP1 in the DA strain of Theiler's murine encephalomyelitis viruses (TMEV) showed that 13 of the first 23 N-terminal amino acids were identical to those in the corresponding protein of encephalomyocarditis virus. There was little similarity to the corresponding VP1 sequences of poliovirus types 1, 2 and 3, coxsackievirus B3, human rhinoviruses 2 and 14, human hepatitis A virus or foot-and-mouth disease virus. These results, as well as serological relationships detected by immunoblotting, suggest that the TMEV are more closely related to the cardioviruses than to the enteroviruses with which they are presently classified. This newly recognized relationship suggests potential for recombinant infectious cDNA studies between TMEV and cardioviruses. PMID- 3034821 TI - Analysis of long-term human cytomegalovirus latency in vitro. AB - Human leukocyte interferon (IFN-alpha) and acyclovir (ACV) have been used to establish human cytomegalovirus (HCMV) latency in infected human embryo lung fibroblast (HEL-F) cells. HCMV latency was maintained for a short interval (less than 9 days) after removal of inhibitors by increasing the incubation temperature. We now report a model system in which HCMV latency has been dramatically extended. HEL-F cells pretreated with IFN-alpha (200 IU/ml) and ACV (300 microM) were infected with a low MOI of HCMV, and treated for 23 days with the same inhibitor combination at 37 degrees. Infectious HCMV and virus antigens were undetectable at the time of inhibitor removal and remained undetectable during continued incubation at 40.5 degrees. A minimum of 0.4% of the cell population, however, contained a virus genome that could be reactivated at the time of inhibitor removal; this value declined to 0.0005% after 77 days at 40.5 degrees. HCMV reactivation was achieved by maintaining the infected cells at 37 degrees after inhibitor removal or by decreasing the incubation temperature to 37 degrees at any time during maintenance at 40.5 degrees. The HCMV genome was analyzed by blot hybridization in latently infected cells 23 days after inhibitor treatment at 37 degrees or 4 days after inhibitor removal at 40.5 degrees. Although many HCMV-unique DNA genomic sequences were retained in HEL-F cultures after 23 days of inhibitor treatment, a significant reduction in retained HCMV sequences occurred after inhibitor removal and temperature shift to 40.5 degrees. The XbaI HCMV DNA fragments retained in the latently infected HEL-F cultures were present at a copy number of at least 0.5 copies per haploid cell genome equivalent. PMID- 3034823 TI - Epidemiology of human rotaviruses in Argentina as determined by RNA genome electrophoresis. AB - Rotavirus gastroenteritis was studied by enzyme-linked immunosorbent assay (ELISA) and by PAGE from April 1983 to April 1985 at the Buenos Aires Ricardo Gutierrez Children's Hospital and at the San Justo Children's Hospital. Based on examination of 576 cases, rotavirus was identified in 109 (18.9%) cases by ELISA and in 99 (17.2%) cases by PAGE. As a diagnostic tool PAGE presented a sensitivity of 90.8% compared with ELISA. Compared with the control SA-11 genome, 84 samples (84.9%) presented a long electropherotype and 15 isolates (15.1%) presented a short electropherotype. We detected 35 different long electropherotypes and report a short electropherotype that did not show genome variation and was responsible for an outbreak in the San Justo Children's Hospital population from September 1983 to July 1984. PMID- 3034824 TI - The strontium-82/rubidium-82 generator. PMID- 3034825 TI - Clinical uses of 82Sr/82Rb generators. PMID- 3034826 TI - A partial decay scheme study of 82Rb and consequences for radiation dose measurements. AB - Measurements of the relative emission rates of the 776 keV gamma ray, the positrons and the K x-rays in the decay of 82Rb have been repeated. This has led to the finding that the gamma ray to positron ratio is 0.1578 +/- 0.0019 and that the 776 keV gamma-ray probability per decay is (15.12 +/- 0.18)%. PMID- 3034827 TI - Essentials of a rubidium-82 generator for nuclear medicine. AB - The use of generator-produced 82Rb for positron emission tomography studies in clinical nuclear medicine requires a number of factors to be considered. These include 82Sr availability, methods of recharging the generator with fresh 82Sr, adequate elution yield of 82Rb, low breakthrough of 82-85Sr, simple and reliable operation of the generator, and delivery of a sterile and pyrogen free eluate of 82Rb. PMID- 3034828 TI - Preparation and evaluation of a hydrous tin(IV) oxide 82Sr/82Rb medical generator system for continuous elution. AB - Hydrous tin(IV) dioxide in the Na+-form appears to be the most efficient inorganic exchanger for a reliable and versatile clinical 82Rb generator. Continuous elution with a commercial physiological NaCl solution yields 82Rb ranging between 10 and 40% at a flow rate as low as 3 to 10 mL/min respectively. At the same time the Sr breakthrough is less than 1.6 10(-6)%/mL. A clinical generator loaded with 100 mCi 82Sr (150 mCi 85Sr) and continuously eluted for 3 min at a typical flow rate of 5 mL/min yields 40 mCi of 82Rb, 8 nCi of 82Sr and 11 nCi of 85Sr. The total absorbed radiation dose for 40 mCi 82Rb administered is primarily due to 82Rb and has been estimated for the three principal target organs as 760 mrad for the kidneys, 520 mrad for the heartwalls and 276 mrad for the lungs. The 82,85Sr contribution to the dosimetry has been shown to be negligible. The absence of radiolysis with generators loaded with high level of 82Sr was demonstrated by the excellent reproductibility of continuous elution properties of the generator during its practical shelf-life estimated to 5-6 weeks of clinical use which would require more than 30 L of eluent. PMID- 3034829 TI - Radioanalysis of 82Rb generator eluates. AB - The activity of 82Rb produced from a 82Sr/82Rb generator is dependent on elution conditions (volume and eluent flow rate) and sampling conditions (time and position of collection). Assays for 82Rb in generator eluates are described using a commercial dose calibrator in a static procedure and a plastic scintillator in a dynamic procedure. Dynamic assays more accurately reflect the 82Rb administered when the eluate is injected directly. Radionuclidic contaminants which may be present with 82Sr are identified and procedures for their measurement are described. PMID- 3034830 TI - Experience with a 82Sr/82Rb generator for clinical use. AB - A 82Sr/82Rb generator system is described which is shown to be suitable for continuous intravenous infusion in man. The breakthrough of the 82Sr parent has been closely monitored and remained less than 18.5 Bq mL-1 of infusate. A method using a 0.05% solution of sodium hypochlorite to disinfect the generator resulted in a sterile and pyrogen free eluate. Recommendations are made for the setting up of the generator to ensure the maintenance of its pharmaceutical integrity. PMID- 3034831 TI - Model for 82Sr/82Rb generator elution profiles: a secondary approach to radioassay/dosimetry. AB - Static (dose calibrator) assays of 82Rb bolus yield erroneous estimates of administered activity by ignoring the asymmetric output of 82Sr/82Rb generators. Though on-line monitoring of elution profile improves accuracy for quantitative purposes, it requires an elaborately shielded detector and associated electronics at patient-study site. Our alternative approach based on mathematical description of the dynamic characteristics of the generator accurately predicts elution profiles over a broad range of flow rates. PMID- 3034832 TI - Small cell carcinoma of the lung in the intensive care unit. PMID- 3034833 TI - Effects of estrogen treatment on the responses of cAMP and sex steroids to hCG by the testes of patients with prostatic cancer. PMID- 3034834 TI - Direct inhibitory effects of 17 beta-estradiol on hCG-stimulated cAMP and testosterone responses of canine testis. PMID- 3034835 TI - Immunolocalization of angiotensin 1 converting enzyme in the human male genital tract by the avidin-biotin-complex method. AB - Immunoreactivity for Angiotensin 1 Converting Enzyme was investigated in a series of 12 fixed and paraffin-embedded normal human genital tract specimens. The Avidin-Biotin-Complex immunoperoxidase method was used with overnight (12 h) incubation with a polyclonal antihuman kidney Angiotensin 1 Converting Enzyme antiserum. All tissues, including testis, different parts of epididymis, ductus deferens, prostate and seminal vesicles, demonstrated a staining pattern. Immunoreactivity was observed on the luminal surface of these epithelia especially on non-motile stereocilia. An intracellular positivity was only observed in spermatids on the acrosomal cap. Besides, an immunologic identity of Angiotensin 1 Converting Enzyme located on the different epithelia of the human male genital tract, on the endothelial cells of vessels and on the proximal tubule brush border of the kidney was observed. PMID- 3034836 TI - Cytochemical demonstration of phosphatases in membrane-recycling structures of endodermal cells. AB - We applied cytochemical procedures to demonstrate the presence of acid and alkaline phosphatase in the visceral yolk-sac endoderm of rats using frozen, aldehyde-fixed tissue with cerium as the capture agent. This procedure allowed more detailed topochemical localization than was possible using unfrozen tissue or with lead as the capture agent. Acid phosphatase was found to be present in lysosomes as well as in a small number of apical canaliculi, which are thought to be recycling structures of the cell membranes in endodermal cells. Reaction products of alkaline phosphatase were observed on the outer surface of apical, lateral, and basal cell membranes. In addition, some apical vacuoles contained alkaline phosphatase, and more apical canaliculi were positive for alkaline phosphatase than for acid phosphatase. However, most of the apical canaliculi were negative for both enzymes. It is suggested that acid and alkaline phosphatase are taken up by different numbers of apical canaliculi during the detachment of apical canaliculi from lysosomes and resorption vacuoles. PMID- 3034837 TI - Serum angiotensin converting enzyme activity in silicosis. PMID- 3034838 TI - Parameters of interaction of a novel monoclonal antibody (33B3.1) with the human IL2-receptors: interrelationship between 33B3.1, anti-Tac, and IL2 binding sites. AB - This report describes the molecular parameters of interaction of a new antibody (33B3.1) with the human membrane RIL2 expressed by ConA-activated T lymphocytes or allogeneic T-cell clones established from a rejected kidney allograft: the 33B3.1 immunoprecipitates a membrane protein of 55000 MW. It inhibits IL2-driven proliferation of activated T cells. This inhibition occurred in the nanomolar range when low concentrations of recombinant IL2 (rec-IL2) were used. The (125I) 33B3.1 binds in a specific way to a single class of receptor sites on activated T cells. The rate constants of association and dissociation at 37 degrees C of the labeled 33B3.1 were k*1 = 12 X 10(5) M-1 s-1 and k*-1 = 7 X 10(-4) s-1, respectively, and its equilibrium dissociation constant was KD = 0.65 nM. Maximal binding capacities were fairly variable among T-cell clones, as high as 300,000 sites/cell for some of them. Competition experiments demonstrate that the 33B3.1 and anti-Tac interact with the RIL2 in a competitive manner, suggesting that they recognize closely associated epitopes on the RIL2. However, the 33B3.1 inhibits the binding of (35S)-recombinant IL2 to its high affinity RIL2 in a noncompetitive way. The 33B3.1 seems therefore to interact with an epitope close but distinct from the IL2 binding site. Our data could suggest either that the 33B3.1 is able to convert high affinity RIL2 towards low affinity conformations or that there is more than one IL2 binding site per molecule of high affinity RIL2. PMID- 3034840 TI - Wilms' tumor complicating pregnancy: report of a case. PMID- 3034839 TI - Rapid elbow flexion in the absence of proprioceptive and cutaneous feedback. AB - Rapid goal-directed movements of elbow flexion were studied in normal human subjects and in patients deprived of proprioceptive and cutaneous feedback. All normal subjects showed a burst of electromyographic (EMG) activity in the extensor muscle (antagonist) that served to arrest the limb precisely in the target zone. The magnitude of this burst co-varied with the magnitude of the initial accelerating burst in the flexor muscle (agonist). In patients, there was a small decelerating burst poorly correlated with the agonist activity. All patients had difficulty to control the amplitude of their movements due to improper adjustment of the size and time of onset of the decelerating burst. It is concluded that the central nervous system can generate a sequence of commands to accelerate and decelerate a limb in the absence of peripheral feedback. However, information from the moving limb is required to adjust the magnitude and time of onset of deceleration. PMID- 3034842 TI - Non-small cell lung cancer: how oncologists want to be treated. AB - One hundred and eighteen Canadian doctors who treat lung cancer were asked how they would wish to be managed if they had non-small cell lung cancer. Four clinical situations were described. The doctors' replies to the open-ended management questions were remarkably consistent in view of their diverse backgrounds. Although opinion was divided as to the role of immediate radiotherapy in inoperable cancer, and the role of post-operative radiotherapy following incomplete surgery, there was little controversy as to the role of chemotherapy. Three per cent of doctors wanted adjuvant chemotherapy after surgery for early disease, 9% wanted chemotherapy for advanced disease confined to the chest and 15% wanted chemotherapy for symptomatic metastatic disease. The implications of these results are discussed in the light of current recommendations for treatment of non-small cell lung cancer in the standard North American textbooks of oncology. PMID- 3034841 TI - Large fraction irradiation with or without misonidazole in advanced non-oat cell carcinoma of the lung: a phase III randomized trial of the RTOG. Radiation Therapy Oncology Group. AB - The Radiation Therapy Oncology Group (RTOG) investigated the use of misonidazole as an hypoxic cell sensitizer in a Phase III prospective randomized trial employing radiotherapy, 600 cGy twice weekly to a total of 3600 cGy with and without misonidazole in the treatment of locally advanced non-metastatic squamous cell, adeno, or large cell carcinoma of the lung. Between January 1980 and July 1983, 117 patients from 21 institutions were enrolled. One-hundred eight patients were evaluable; 53 in the combined treatment arm and 55 in the radiation alone arm. Grade 3 or worse complications associated with radiation occurred in 17% of patients. Esophageal toxicity accounted for the majority of complications. Two (4%) patients in the radiotherapy plus misonidazole group experienced grade 3 peripheral neurotoxicity. Complete or partial responses were produced in 58% of the patients with radiotherapy alone and 36% of those treated with radiotherapy plus misonidazole (p = 0.08). At the time of first progression, over 50% of the patients had persistent local disease. Median survival was 7 months regardless of treatment. Misonidazole in the dose and schedule employed did not enhance the effect of radiotherapy on either local tumor control or overall survival in patients with advanced lung cancer. PMID- 3034843 TI - Evidence for age-dependent changes in Sertoli cell androgen receptor concentration. AB - Cytosol and nuclear receptor concentrations in Sertoli cells isolated from the testes of 15-, 25-, and 35-day-old rats were measured using hydroxylapatite separation procedures. In these cells the mean Kd of the cytosol receptor for methyltrienolone (3H-R1881) ranged between 2.3 and 2.9 nM, and the concentration of cytosol androgen receptor per mg Sertoli cell DNA increased over the 15-to 35 day age interval. However, when the data were expressed per mg cytosol protein, no increase was observed. The increase in receptor concentration per mg DNA paralleled the increase in cytosol protein/DNA ratio. The concentration of androgen receptor per mg DNA in nuclear extracts also increased with age. Consequently, total Sertoli cell androgen receptor increases over the time interval in which meiosis is first completed in the testis. PMID- 3034844 TI - K-13, a novel inhibitor of angiotensin I converting enzyme produced by Micromonospora halophytica subsp. exilisia. I. Fermentation, isolation and biological properties. AB - A novel inhibitor of angiotensin I converting enzyme (ACE), designated K-13, was isolated from the culture broth of Micromonospora halophytica subsp. exilisia K 13. K-13 inhibited ACE non-competitively when hippuryl-L-histidyl-L-leucine was used as a substrate. The inhibition constant (Ki) was 0.349 microM. K-13 hardly inhibited carboxypeptidase A, trypsin, alpha-chymotrypsin, leucine aminopeptidase, and aminopeptidase B even at a level of 61 microM. When K-13 was administered intravenously to rats, it inhibited the pressor response to angiotensin I. PMID- 3034845 TI - K-13, a novel inhibitor of angiotensin I converting enzyme produced by Micromonospora halophytica subsp. exilisia. II. Structure determination. AB - The structure of K-13, a potent inhibitor of angiotensin I converting enzyme (ACE), was determined to be a cyclic dipeptide composed of tyrosine and an unusual diamino dicarboxylic acid, isodityrosine, by spectral and chemical studies of K-13 and its derivatives. PMID- 3034846 TI - WF-10129, a novel angiotensin converting enzyme inhibitor produced by a fungus, Doratomyces putredinis. AB - WF-10129 is an angiotensin converting enzyme (ACE) inhibitor produced by Doratomyces putredinis. IC50 of the compound is 1.4 X 10(-8) M for the ACE activity. WF-10129 was purified from cultured filtrate by successive ion exchange chromatography and HPLC. The chemical structure 1 was elucidated on the basis of spectroscopic and chemical evidence. The compound is a dipeptide composed of L tyrosine and a novel amino acid. WF-10129 inhibits the pressor response of angiotensin I when administered intravenously at 0.3 mg/kg in rats. PMID- 3034848 TI - Synergistic resistance mechanisms in Pseudomonas aeruginosa. PMID- 3034847 TI - Protective effect of forphenicinol, a low molecular weight immunomodifier, against infection with Pseudomonas aeruginosa in mice and its mechanisms. AB - The oral administration of forphenicinol increased the survival rate of both normal and immunodepressed mice intraperitoneally or intratracheally infected with clinically isolated strains of Pseudomonas aeruginosa. The therapeutic effect of amikacin on intraperitoneal infection with P. aeruginosa was enhanced by combined use with forphenicinol. Forphenicinol did not enhance the bactericidal activity of polymorphonuclear cells (PMN) towards P. aeruginosa in vitro, but enhanced it in vivo. In vitro study indicated that the macrophages taken from mice treated with forphenicinol or the cultured supernatant of these macrophages enhanced the bactericidal activity of PMN. The protective effect of forphenicinol against P. aeruginosa infection was thus suggested to be due to macrophage activation followed by the enhancement of the bactericidal activity of PMN. PMID- 3034849 TI - In-vitro activity of LY 146032, a novel cyclic lipopeptide, alone and in combination with gentamicin or tobramycin against enterococci. AB - The in-vitro activity of LY 146032, a novel cyclic lipopeptide and the effect of the combination of LY 146032 and gentamicin or tobramycin against 25 strains of enterococci isolated from blood cultures were studied. All strains of enterococci were inhibited by less than or equal to 2 mg/l LY 146032. The minimal inhibitory concentrations and minimal bactericidal concentrations were within one dilution. In the time-kill study, there was slow bactericidal activity. Complete killing of 10(7) cfu/ml of enterococci at 48 h by LY 146032 alone occurred with two strains. There was synergism between LY 146032 and gentamicin or tobramycin; complete killing at 24 h and 48 h occurred with many strains. No antagonism was demonstrated. PMID- 3034850 TI - Clinical efficacy and safety of sulbactam/ampicillin in patients suffering from chronic liver disease. AB - Sulbactam is a new beta-lactamase inhibitor with pharmacokinetic characteristics in humans similar to those of ampicillin. A total of 41 patients hospitalized in the Clinic of Infectious Diseases, University of Naples, for chronic liver diseases, were treated with sulbactam/ampicillin (ratio 1:2) for urinary, respiratory, biliary tract or soft tissue infections. Sulbactam/ampicillin was administered im or iv at a dosage of 3-9 g/day depending on the site and severity of the infection. All the patients treated with sulbactam/ampicillin had clinical signs and symptoms of infection, and all the organisms isolated were sensitive to sulbactam/ampicillin (MIC less than 16 mg/l). For both Gram-positive and Gram negative bacteria the sulbactam/ampicillin MICs were much lower than the ampicillin MICs. In agreement with the favourable in-vitro results, we observed good therapeutic efficacy. 85% of the patients recovered or improved within a few days of therapy, with no clinical relapses, and in 81% of the infections the responsible bacteria were completely eradicated. We observed a low number of side effects (3/41 oral candidosis; 3/41 pain at the im injection site) and no change in the blood chemistry tests. PMID- 3034851 TI - Growth and hepatospecific gene expression of human hepatoma cells in a defined medium. AB - The production of albumin, alpha-fetoprotein (AFP), and alpha-1 antitrypsin has been compared among human hepatoma cells cultured in medium containing serum, medium containing hormones and growth factors, and a basal medium containing selenium as the only supplement. Growth is sustained in all three media, and the expression of all three proteins was maintained for over 4 mo. in the various media. However, the quantitative production of albumin and AFP were dramatically different in the three media. Two hormones, insulin and triiodothyronine, influenced the level of secreted proteins. Triiodothyronine increases the amount of secreted albumin whereas insulin at 10 micrograms/ml reduced the level of total secreted protein. PMID- 3034852 TI - Parotid gland salivary secretion in Tourette's syndrome and attention deficit disorder: a model system for the study of neurochemical regulation. PMID- 3034853 TI - Gene expression in Zymomonas mobilis: promoter structure and identification of membrane anchor sequences forming functional lacZ' fusion proteins. AB - We have described a procedure for the isolation of lacZ' fusion genes which contain anchor sequences conferring membrane association. This method was used to isolate fragments of DNA from Zymomonas mobilis which contain promoter activity and amino-terminal sequences. The sequences and transcriptional initiation sites of three of these were compared. Both Escherichia coli and Z. mobilis recognized similar regions of DNA for transcriptional initiation. Five to eight consecutive hydrophobic amino acids in the amino terminus served to anchor these hybrid proteins to the membrane in both E. coli and Z. mobilis. General features observed in the Z. mobilis fragments included partial sequence homology with the 35 region sequence of E. coli, repetitive and palindromic A + T-rich regions preceding and adjoining the -10 region, a sequence resembling the consensus sequence of E. coli in the -10 region, and a potential ribosomal-binding site (AGGA) 8 to 12 bases upstream from an in-frame start codon. The level of expression of fusion proteins was generally higher in E. coli than in Z. mobilis. This higher level of expression in E. coli may result from multiple sites of transcriptional initiation and higher plasmid copy number. PMID- 3034854 TI - Spectinomycin resistance and associated DNA amplification in Streptomyces achromogenes subsp. rubradiris. AB - Streptomyces achromogenes subsp. rubradiris plated at low density on 1,000 micrograms of spectinomycin per ml initially produces slow-growing, bald colonies from which arise, in a spatially and temporally random fashion, foci of rapidly growing aerial mycelium-forming cells whose DNA contains an approximately 200- to 300-fold amplification of an 8-kilobase (kb) sequence. This sequence was cloned in Escherichia coli on pBR322 and physically characterized. It was separately cloned also in Streptomyces lividans as a BglII fragment and shown to impart high level resistance to spectinomycin in an orientation-independent manner when present in either the high-copy-number vector pIJ702 or the unit-copy-number vector pIJ943. A spectinomycin resistance determinant was shown to reside on a 1.7-kb SphI-BglII subfragment. Analysis of Southern blots of restriction enzyme digests of wild-type S. achromogenes DNA probed with the labeled 8-kb DNA sequence resulted in the identification and subsequent cloning in S. lividans of a 10.4-kb BamHI fragment which probably includes the complete 8.8-kb amplifiable unit of DNA. This unit is present in wild-type S. achromogenes and in the initially slow-growing, bald colonies arising on 1,000 micrograms of spectinomycin per ml as a single copy. It carries two 0.8-kb direct repeats at its termini as well as the spectinomycin resistance determinant close to one of these termini. About 5% of protoplast regenerants from wild-type S. achromogenes and 77% of protoplast regenerants from the rapidly growing strains lost both the ability to grow on spectinomycin at 10 micrograms/ml and the sequences that hybridize with the 8-kb probe DNA. The 1.7-kb Bg/II-SphI resistance fragment, when introduced via the vector pIJ702 into an S. achromogenes strain sensitive to 10 microgram of spectinomycin per ml, permitted its vigorous growth on 1,000 micrograms of the antibiotic per ml. PMID- 3034855 TI - Initial catabolism of aromatic biogenic amines by Pseudomonas aeruginosa PAO: pathway description, mapping of mutations, and cloning of essential genes. AB - Pseudomonas aeruginosa PAO1 was able to utilize several aromatic biogenic amines as sole sources of carbon or nitrogen. These included the phenethylamines tyramine and dopamine and the phenethanolamines octopamine, synephrine, and norepinephrine. Initial catabolism of the phenethylamines was mediated by a membrane-bound tyramine dehydrogenase which produced 4-hydroxyphenylacetaldehyde (4HPAL) with tyramine as the substrate. The enzyme was induced by growth with both classes of amines. Initial catabolism of octopamine (except when present as the sole source of carbon and nitrogen) was mediated by a soluble enzyme with activity against the phenethanolamines but not against tyramine or dopamine. The product of the reaction with octopamine as substrate was also 4HPAL. Addition of NAD to reaction mixtures yielded 4-hydroxyphenylacetic acid and NADH. These activities, octopamine hydrolyase and 4-HPAL dehydrogenase (measured as a combined activity, OCAH-4HPALDH), were only induced by growth with phenethanolamines. However, the combined activities were not observed in extracts from cells grown with octopamine as the sole source of carbon and nitrogen, suggesting that an alternate pathway is used under this growth condition. Two independently isolated mutant strains were unable to utilize tyramine as a sole source of carbon or nitrogen. These mutants were also unable to utilize dopamine but grew at wild-type rates on the phenethanolamines. The mutations were mapped at about 70 min on the PAO1 chromosome with the chromosome-mobilizing plasmid R68.45, and both were linked to the catA1, mtu-9002, tyu-9009, and puuE mutations. DNA complementing both of the mutations was cloned on a single BamHI fragment approximately 13.8 kilobase pairs in length. Analysis of a subcloned fragment showed that the two mutations were in different genes. PMID- 3034856 TI - Rhizobium meliloti ntrA (rpoN) gene is required for diverse metabolic functions. AB - We report the identification and cloning of an ntrA-like (glnF rpoN) gene of Rhizobium meliloti and show that the R. meliloti ntrA product (NtrA) is required for C4-dicarboxylate transport as well as for nitrate assimilation and symbiotic nitrogen fixation. DNA sequence analysis showed that R. meliloti NtrA is 38% homologous with Klebsiella pneumoniae NtrA. Subcloning and complementation analysis suggested that the R. meliloti ntrA promoter lies within 125 base pairs of the initiation codon and may be constitutively expressed. PMID- 3034857 TI - Four plasmid genes are required for colicin V synthesis, export, and immunity. AB - The colicin V production and immunity genes were isolated from plasmid pColV-K30. A HindIII-to-SalI fragment of 9.4 kilobases was cloned into the compatible vectors pBR322 and pACYC184. Mutants defective in colicin production were generated by Tn5 insertions and by constructing deletions in vitro. Physical analysis of these mutations identified a 4.4-kilobase region of this DNA which contains all the plasmid genes (cva) needed for the production of colicin V. The colicin V immunity determinant (cvi) is in a 700-base-pair fragment located within one end of this region. Complementation tests identified three genes, called cvaA, cvaB, and cvaC, required for colicin production. Analysis of the proteins labeled in minicells harboring various Tn5 insertions allowed us to identify protein products for the cvaA and cvaC genes. Mutations in cvaA and cvaB eliminated colicin activity in culture supernatants, but not within the cells. Mutations in cvaC, however, eliminated all detectable activity. From these results we conclude that the cvaC gene codes for the structural gene for colicin V, while cvaA and cvaB are apparently needed for the normal export of the colicin. PMID- 3034858 TI - Role of oxygen radicals in the phototoxicity of tetracyclines toward Escherichia coli B. AB - Photoillumination of tetracycline derivatives with low-intensity (320- to 400-nm) light and visible light generated superoxide, observed as the reduction of ferricytochrome c. The rate of reduction was dependent on the tetracycline concentration and on the derivative being examined, with doxycycline greater than or equal to demeclocycline greater than tetracycline greater than oxytetracycline. Tetracycline-mediated cytochrome c reduction was oxygen dependent and inhibited up to 70% by superoxide dismutase. Illuminated tetracyclines were lethal to Escherichia coli B incubated in a glucose minimal medium containing chloramphenicol. This lethality was light dependent, oxygen dependent, and dependent on the concentration of tetracycline. Kill rates also varied according to the derivative under study, with doxycycline greater than or equal to demeclocycline greater than tetracycline greater than oxytetracycline. The addition of superoxide dismutase and catalase to the incubation medium partially protected E. coli B against the light-dependent lethality. Preinduction of intracellular superoxide dismutase and catalase substantially protected E. coli B against the phototoxicity of tetracyclines. Iron EDTA augmented the phototoxicity of tetracyclines, while diethylenetriaminepentaacetic acid protected against their lethality. Hydroxyl radical scavengers also conferred protection against tetracycline phototoxicity. The extent of protection was in order of the in vitro reactivity of the scavengers with the hydroxyl radical. These results indicate that superoxide, hydrogen peroxide, and the hydroxyl radical are generated by illuminated tetracyclines and are molecular agents of tetracycline phototoxicity in E. coli B. PMID- 3034859 TI - Two conjugation systems associated with Streptococcus faecalis plasmid pCF10: identification of a conjugative transposon that transfers between S. faecalis and Bacillus subtilis. AB - The tetracycline resistance plasmid pCF10 (58 kilobases [kb]) of Streptococcus faecalis possesses two separate conjugation systems. A 25-kb region of the plasmid (designated TRA) was shown previously to determine pheromone response and conjugation functions required for transfer of pCF10 between S. faecalis cells (P. J. Christie and G. M. Dunny, Plasmid 15:230-241, 1986). When S. faecalis cells were mixed with Bacillus subtilis in broth, tetracycline resistance was transferred from S. faecalis. The tetracycline-resistant B. subtilis cells contained a 16-kb region of pCF10 (distinct from TRA) that carried the tetracycline resistance determinant (Tetr). This Tetr element was found to transfer between S. faecalis and B. subtilis strains in the absence of plasmids. Genetic and molecular techniques were used to establish locations of the element at several different sites on the B. subtilis chromosome. The Tetr element could be transferred in filter matings from B. subtilis to S. faecalis strains and between recombination-proficient and -deficient S. faecalis strains in the absence of any plasmid DNA. The transfer required direct cell-to-cell contact and was not inhibited by DNase. The Tetr element was shown to transpose from the S. faecalis chromosome to various locations within the hemolysin plasmid pAD1. Together, the data indicate that the Tetr element, termed transposon Tn925, is very similar to the conjugative transposon Tn916 in both structure and function. A derivative of Tn925, containing transposon Tn917 inserted into a site approximately 3 kb from one end, exhibited elevated transfer frequencies and may provide a useful means for delivering Tn917 by conjugation into various gram positive species. PMID- 3034860 TI - Beta-glucoside (bgl) operon of Escherichia coli K-12: nucleotide sequence, genetic organization, and possible evolutionary relationship to regulatory components of two Bacillus subtilis genes. AB - Wild-type Escherichia coli cells are unable to grow on beta-glucosides. Spontaneous mutants arise, however, which are able to utilize certain aromatic beta-glucosides such as salicin or arbutin as carbon sources, revealing the presence of a cryptic operon called bgl. Mutations activating the operon map within (or close to) the promoter region of the operon and are due to the transposition of an IS1 or IS5 insertion element into this region. This operon was reported to consist of three genes coding for a phospho-beta-glucosidase, a specific transport protein (enzyme IIBgl), and a positively regulating protein. We have defined the extent and location of three structural genes, bglC, bglS, and bglB, and have determined their DNA sequence. The amino acid sequences deduced from the open reading frames together with deletion and subcloning analyses suggest that the first gene, bglC, codes for the regulatory protein, the second, bglS, codes for the transport protein, and the third, bglB, for phospho beta-glucosidase. A fourth gene may exist which codes for a product of unknown function. We discuss structural features of the DNA sequence which may bear on the regulation of the operon. Homologies to sequences preceding the gene for an excreted levansucrase of Bacillus subtilis, which are known to be involved in the regulation of this gene, and to sequences preceding the gene for an excreted beta endoglucanase of B. subtilis, for which data pertaining to regulation are not yet available, suggest a close evolutionary relationship among the regulatory components of all three systems. PMID- 3034861 TI - Identification, mapping, and cloning of the gene encoding cyanase in Escherichia coli K-12. AB - The gene in Escherichia coli for cyanase, designated cynS, was localized to a BglII restriction site approximately 1.7 kilobases from the lacA end of the lac operon. The gene was cloned into the pUC13 vector. Maxicell analysis of plasmid encoded proteins confirmed that the BglII site is in the region encoding the structural gene for cyanase. Cyanase-deficient strains had increased sensitivity to cyanate and were not able to use cyanate as a nitrogen source. PMID- 3034862 TI - Mode of initiation of constitutive stable DNA replication in RNase H-defective mutants of Escherichia coli K-12. AB - The alternative pathway of DNA replication in rnh mutants of Escherichia coli can be continuously initiated in the presence of chloramphenicol, giving rise to constitutive stable DNA replication (cSDR). We conducted a physiological analysis of cSDR in rnh-224 mutants in the presence or absence of the normal DNA replication system. The following results were obtained. cSDR allowed the cells to grow in the absence of the normal replication system at a 30 to 40% reduced growth rate and with an approximately twofold-decreased DNA content. cSDR initiation was random with respect to time in the cell cycle as well as choice of origins. cSDR initiation continued to increase exponentially for more than one doubling time when protein synthesis was inhibited by chloramphenicol. cSDR initiation was inhibited during amino acid starvation in stringent (relA+) but not in relaxed (relA1) strains, indicating its sensitivity to ppGpp. cSDR initiation was rifampin sensitive, demonstrating that RNA polymerase was involved. cSDR functioned in dnaA+ rnh-224 strains parallel to the normal oriC+ dnaA+-dependent chromosome replication system. PMID- 3034863 TI - Expression and regulation of a Vibrio alginolyticus sucrose utilization system cloned in Escherichia coli. AB - A halotolerant collagenolytic Vibrio alginolyticus strain isolated from salted hides had intracellular sucrase activity and did not secret sucrase into the medium. The strain actively transported sucrose by a sucrose-inducible, Na+ independent process. A 10.4-kilobase DNA fragment of V. alginolyticus DNA was cloned into Escherichia coli. The recombinant E. coli(pVS100) could utilize sucrose as a sole carbon source. In contrast to V. alginolyticus, the recombinant E. coli produced both intra- and extracellular sucrase activities. Up to 20% of the total sucrase activity was in the supernatant. Sucrase synthesis in E. coli(pVS100) was inducible and was subject to glucose repression, which was relieved by cyclic AMP. Sucrose was actively transported by a sucrose-inducible, Na+-independent system in E. coli(pVS100). Sucrose uptake was inhibited by the addition of a proton conductor. The maximum velocity and apparent Km values of sucrose uptake for the V. alginolyticus strain and E. coli(pVS100) were 130 nmol/mg of protein per min and 50 microM and 6 nmol/mg of protein per min and 275 microM, respectively. PMID- 3034865 TI - Inhibition of hydrogenase synthesis by DNA gyrase inhibitors in Bradyrhizobium japonicum. AB - Derepression of an uptake hydrogenase in Bradyrhizobium japonicum is dependent on a microaerophilic environment. Addition of DNA gyrase inhibitors during depression of hydrogenase specifically prevented expression of the hydrogenase enzyme. Antibodies to individual hydrogenase subunits failed to detect the protein after derepression in the presence of inhibitors, although there was no general inhibition of protein synthesis. The general pattern of proteins synthesized from 14C-labeled amino acids during derepression was not significantly different whether proteins were labeled in the presence or in the absence of gyrase inhibitors. In contrast, if transcription or translation was inhibited by addition of inhibitors of those functions, virtually no proteins were labeled during derepression. This indicated that most of the 14C-labeled proteins were synthesized de novo during derepression, synthesis of most proteins was unaffected by gyrase inhibitors, and the dependence of hydrogenase synthesis on gyrase activity was a specific one. PMID- 3034866 TI - Functional genes for cellobiose utilization in natural isolates of Escherichia coli. AB - The genes for utilization of cellobiose are normally cryptic in both laboratory strains and natural isolates of Escherichia coli. A survey of natural isolates of E. coli reveals that functional genes for cellobiose utilization, while rare, are present. The fraction of E. coli that utilized cellobiose ranged from less than 0.01% in human fecal samples to 7% in fecal samples obtained from horses. Samples obtained from sheep, cows, dogs, and pigs contained 0.1 to 0.5% cellobiose positive E. coli. Neither the previously identified cel genes nor the bgl genes from E. coli K-12 were expressed during growth on cellobiose by any of the 14 naturally occurring Cel+ isolates that were tested. All of the naturally occurring Cel+ isolates possessed a cel operon, but all were deleted for the major portion of the bgl operon. The functional cel+ genes from these natural isolates differed from the mutationally activated cel+ genes obtained in earlier studies in that (i) the mutationally activated cel+ genes were temperature sensitive, while the functional genes were not, and (ii) transport of cellobiose was inducible in the strains carrying functional cel+ genes, while it was expressed constitutively in strains carrying mutationally activated genes. PMID- 3034864 TI - Cloning and characterization of elastase genes from Pseudomonas aeruginosa. AB - A gene bank was constructed from Pseudomonas aeruginosa PAO1 and used to complement three P. aeruginosa elastase-deficient strains. One clone, pRF1, contained a gene which restored elastase production in two P. aeruginosa isolates deficient in elastase production (PA-E15 and PAO-E105). This gene also encoded production of elastase antigen and activity in Escherichia coli and is the structural gene for Pseudomonas elastase. A second clone, pHN13, contained a 20 kilobase (kb) EcoRI insert which was not related to the 8-kb EcoRI insert of pRF1 as determined by restriction analysis and DNA hybridization. A 2.2-kb SalI HindIII fragment from pHN3 was subcloned into pUC18, forming pRB1822-1. Plasmid pRB1822-1 restored normal elastolytic activity to PAO-E64, a mutant for elastase activity. Clones derived from pHN13 failed to elicit elastase antigen or enzymatic activity in E. coli. PMID- 3034867 TI - Genetic transformation in the methanogen Methanococcus voltae PS. AB - Mutations causing requirements for histidine, purine, and vitamin B12 were obtained in strain PS of Methanococcus voltae (archaebacteria) upon irradiation with UV or gamma rays. The first two mutations were shown to revert at low frequencies and were used to demonstrate the occurrence of transformation with homologous, wild-type DNA. The transformation rates obtained for these presumably chromosomal markers were in the range of 2 to 100 transformants per microgram of DNA. Mutants resistant to 2-bromoethanesulfonate and to 5-methyl-DL-tryptophan were also isolated. PMID- 3034869 TI - Evidence that colicin X is microcin B17. AB - The DNA replication inhibitor microcin B17 is a peptide antibiotic produced by Escherichia coli strains carrying plasmid pMccB17. Here we present evidence that antibiotic activities previously named colicin X are probably identical to microcin B17. Our results include comparison of the conditions of production of the antibiotics, their mode of action, cross-immunity of producer strains, and cross-resistance of resistant mutants. Plasmids encoding colicin X have been identified and shown to have a region of significant homology with the microcin B17-producing region of pMccB17 DNA. PMID- 3034868 TI - Genetic and physical analyses of a cluster of genes essential for xanthan gum biosynthesis in Xanthomonas campestris. AB - Xanthomonas campestris produces copious amounts of a complex exopolysaccharide, xanthan gum. Nonmucoid mutants, defective in synthesis of xanthan polysaccharide, were isolated after nitrosoguanidine mutagenesis. To isolate genes essential for xanthan polysaccharide synthesis (xps), a genomic library of X. campestris DNA, partially digested with SalI and ligated into the broad-host-range cloning vector pRK293, was constructed in Escherichia coli. The pooled clone bank was conjugated en masse from E. coli into three nonmucoid mutants by using pRK2013, which provides plasmid transfer functions. Kanamycin-resistant exconjugants were then screened for the ability to form mucoid colonies. Analysis of plasmids from several mucoid exconjugants indicated that overlapping segments of DNA had been cloned. These plasmids were tested for complementation of eight additional nonmucoid mutants. A 22-kilobase (kb) region of DNA was defined physically by restriction enzyme analysis and genetically by ability to restore mucoid phenotype to 10 of the 11 nonmucoid mutants tested. This region was further defined by subcloning and by transposon mutagenesis with mini-Mu(Tetr), with subsequent analysis of genetic complementation of nonmucoid mutants. A region of 13.5 kb of DNA was determined to contain at least five complementation groups. The effect of plasmids containing cloned xps genes on xanthan gum synthesis was evaluated. One plasmid, pCHC3, containing a 12.4-kb insert and at least four linked xanthan biosynthetic genes, increased the production of xanthan gum by 10% and increased the extent of pyruvylation of the xanthan side chains by about 45%. This indicates that a gene affecting pyruvylation of xanthan gum is linked to this cluster of xps genes. PMID- 3034871 TI - The COOH-terminal E-F hand structure of calcium-activated neutral protease (CANP) is important for the association of subunits and resulting proteolytic activity. AB - High-Ca2+-requiring calcium-activated neutral protease (mCANP), a dimeric enzyme composed of large (Mr = 80,000) and small (Mr = 28,000) subunits, is resistant to carboxypeptidase Y (CPase Y) in the absence of NaSCN. In the presence of 0.2 M NaSCN, CPase Y digested mCANP, one or two amino acids being released from the COOH-termini of the large and small subunits, but no change occurred in the activity of the digested mCANP. In the presence of 1 M NaSCN, 8-10 amino acids were released from the subunits by CPase Y, and the COOH-terminal potential Ca2+ binding sites of both subunits were destroyed. On digestion under these conditions, mCANP lost the ability to form a complex, and the proteolytic activity was not recovered even when the digested subunits were mixed with native subunits. These results suggest that the COOH-terminal regions of the two subunits of mCANP, which constitute the helical portions of the COOH-terminal E-F hand structures in both subunits, are essential for the subunit association and resulting proteolytic activity. PMID- 3034870 TI - The methylamine oxidizing system of Pseudomonas AM1 reconstituted with purified components. AB - The electron transport system coupled to the oxidation of methylamine in Pseudomonas AM1 was investigated by reconstituting it from the highly purified components. A mixture of methylamine dehydrogenase, cytochrome cH and cytochrome c oxidase (= cytochrome aa3) actively oxidized methylamine (161 mol of O2 consumed/mol of heme a of cytochrome c oxidase X min). In this system, addition of amicyanin did not affect the oxygen consumption rate. The oxygen consumption rate of the cell-free extract prepared from the cells cultivated in a copper deficient medium was directly proportional to the amount of amicyanin added, and extrapolation to zero copper concentration gave a value of 28 mol of O2 consumed/mol of heme a of cytochrome c oxidase X min. These results suggest that methylamine oxidation in the bacterium can occur at least to some extent without participation of amicyanin. PMID- 3034873 TI - The tetranuclear manganese complex of Photosystem II. AB - A polynuclear manganese complex functions in Photosystem II both to accumulate oxidizing equivalents and to bind water and catalyze its four-electron oxidation. Recent electron paramagnetic resonance (EPR) spectroscopic studies of the manganese complex show that four manganese ions are required to account for its magnetic properties. The exchange couplings between manganese ions in the S2 state are characteristic of a Mn4O4 "cubane"-like structure. Based on this structure for the manganese complex in the S2 state, as well as a consideration of the known properties of the manganese complex in Photosystem II and the coordination chemistry of manganese, structures are proposed for the five intermediate oxidation states of the manganese complex. A molecular mechanism for the formation of an O-O bond and the displacement of O2 from the S4 state is suggested. PMID- 3034875 TI - Spectroscopic studies on the interaction of phosphate with uteroferrin. AB - The effect of phosphate on the binuclear iron center of pink (reduced) uteroferrin was examined by magnetic resonance and optical spectroscopy. The purple (oxidized) protein, which contains 1 mol of tightly bound phosphate per mol of enzyme at isolation, does not give rise to a 31P NMR signal. Phosphate binding to phosphate-stripped pink uteroferrin is indistinguishable from that in the native purple phosphoprotein. As measured by EPR and optical spectroscopy, the rate of reaction between phosphate and pink uteroferrin is pH-dependent, decreasing as the pH increases. Phosphate is capable of binding to the reduced protein between pH 3 and 7.8, resulting in formation of the purple uteroferrin phosphate complex. Evans susceptibility measurements at pH 4.9 indicate that the EPR silent species with a maximum absorption at 535 nm, generated upon phosphate addition to pink uteroferrin, is diamagnetic. Moreover, phosphate causes disappearance of the hyperfine-shifted resonances in the 1H NMR spectra of the reduced protein. We therefore have not been able to identify the paramagnetic "purple reduced enzyme-phosphate complex" reported by Pyrz et al. (Pyrz, J. W., Sage, J. T., Debrunner, P. G., and Que, Jr., L. (1986) J. Biol Chem. 261, 11015 11020) using Mossbauer spectroscopy and dithionite-reduced 57Fe-reconstituted uteroferrin. Our present data with native unmodified enzyme are in accord with our earlier results (Antanaitis, B. C., and Aisen, P. (1985) J. Biol. Chem. 260, 751-756) and with the results of Burman et al. (Burman, S., Davis, J. C., Weber, M. J., and Averill, B. A. (1986) Biochem. Biophys. Res. Commun. 136, 490-497) on bovine spleen phosphatase, suggesting that phosphate binding to reduced protein rapidly induces oxidation of the binuclear iron center. PMID- 3034874 TI - The active form of tumor necrosis factor is a trimer. AB - Natural human and recombinant human and murine tumor necrosis factors (TNF) were fractionated by gel filtration chromatography on Sephadex G-75. The active form of TNF was identified by its inhibitory activity in receptor binding assays with HeLa cells and was eluted as a protein of Mr approximately 55,000. Radioiodinated human and murine TNF were fractionated by gel filtration into a major peak of Mr approximately 55,000, corresponding to a trimer, and a minor peak of Mr approximately 17,000, corresponding to a monomer. Binding assays showed that the timer was at least 8-fold more active than the monomer. The human TNF partially dissociated into monomers upon addition of the nonionic detergent Triton X-100. Isolated monomers showed low binding affinity (KD = 70 nM) and reduced cytotoxicity, whereas trimers showed high binding affinity (KD = 90 pM) and cytotoxicity. When 125I-TNF was bound to cells, no release of monomer was detectable, suggesting that the trimer could directly bind to cellular receptors without dissociating into subunits. Further evidence for such binding was obtained by cross-linking 125I-TNF trimers with bis[2 (succinimidooxycarbonyloxy)ethyl]sulfone. These trimers were bound to HeLa cells, could be dissociated from cellular receptors, and elicited a cytotoxic response. These results show that trimers, whether native or cross-linked, bind to receptors and are the biologically active form of TNF. PMID- 3034872 TI - Determination of the orientation of membrane vesicles derived from mitochondria. AB - Membrane vesicles of physiological as well as inverted orientation can be isolated from mitochondria. The presence of these vesicles in a membrane can be determined and quantitated by determining the differences between the two vesicle types in terms of rates of NADH oxidation, rates of oxidation of tricarboxylate cycle intermediates, rates of ATP hydrolysis and sensitivity to inhibitors, stimulation of respiration by exogenous cytochrome c, inhibition of respiration by polycationic proteins, and visualization of the ATPase by electron microscopy. Procedures to isolate the two membrane types and characteristics of homogeneously oriented preparations are described. Differences in data obtained with homogeneous vesicle preparations and with vesicles of mixed orientation are illustrated. Nonhomogeneously oriented preparations can be enriched in the desired vesicular type by the use of immunoprecipitation, affinity chromatography, and differential centrifugation. The concept of a hybrid vesicle containing oppositely oriented regions is not supported by experimental data. PMID- 3034876 TI - Structural analysis of the locus containing the human C-reactive protein gene and its related pseudogene. AB - The gene for human C-reactive protein (CRP) is mapped within a 34-kilobase pair genomic DNA segment identified by chromosome walking through overlapping DNA fragments cloned into a lambda phage library. Within 16 kilobase pairs upstream and downstream of the locus for the authentic CRP gene, only one other sequence homologous to that for CRP could be found. Sequencing analysis indicates this sequence to be a pseudogene with 50-80% region-specific homology. Comparison of the authentic CRP gene cloned from genomic DNA libraries independently prepared from three patients indicates no difference in the 5' and 3' flanking region, promoter region, or coding sequence. Only a polymorphism in the length of the poly(GT) stretch located in the intron is observed. There appears to be only one gene locus and copy per haploid chromosome for the authentic CRP gene and its pseudogene. PMID- 3034878 TI - Proteolysis of human C-reactive protein produces peptides with potent immunomodulating activity. AB - We have studied the ability of human C-reactive protein to modulate the immune response in vitro. Whereas native C-reactive protein did not induce phagocytic leukocytes to chemotax or to produce superoxide, treatment of purified C-reactive protein with human neutrophil-derived acid proteases produced substances with potent effects on leukocyte function. Close examination of the primary structure of human C-reactive protein revealed three regions evenly distributed throughout the protein each of which contain peptide sequences closely resembling the amino acid sequence of the immunomodulator peptide tuftsin, Thr-Lys-Pro-Arg. We have synthesized the three peptides which include Thr-Lys-Pro-Leu ([Leu4]tuftsin), Gly Lys-Pro-Arg ([Gly1]tuftsin), and Thr-Lys-Pro-Gln ([Gln4]tuftsin) and assayed them for biological activity. The three synthetic peptides were found to stimulate phagocytic leukocytes to chemotax, produce superoxide, and induce mononuclear cells to produce interleukin 1 in vitro at concentrations similar to those concentrations required for tuftsin to induce these phenomena. These results support a potentially important role for C-reactive protein as a possible immunomodulator during inflammation. PMID- 3034879 TI - Salt-induced activation of 1,25-dihydroxyvitamin D3 receptors to a DNA binding form. AB - In this report we examine the DNA-cellulose binding and sedimentation properties of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) receptors from rat intestine and cultured human mammary cancer cells (MCF-7) extracted in nonactivating (low salt) buffers. Receptors prepared in hypotonic buffer had low DNA binding (13%) compared to receptors extracted with 0.3 M KCl (50%). Treatment of low salt receptor preparations with KCl significantly increased (approximately 3-fold) DNA binding (activation), demonstrating that receptors can be "activated" in vitro. Activated receptors eluted from DNA-cellulose at 0.18 M KCl. Sedimentation analysis followed by DNA-cellulose binding indicated that activated receptors are approximately 3.2 S and unactivated receptors 5.5 S in size. These results suggest that dissociation of an aggregated moiety may lead to receptor activation. Treatment of unactivated receptor with RNase did not alter DNA binding or sedimentation properties of the aggregated receptor. Treatment of unactivated receptor complexes with heat did not increase DNA binding, and molybdate did not block subsequent salt activation. In summary these results suggest that 1,25(OH)2D3 receptors undergo a salt-induced activation step similar to that described for other steroid receptor systems. However, 1,25(OH)2D3 receptors differ from other steroid receptors in not exhibiting heat activation nor having salt activation blocked by molybdate. PMID- 3034877 TI - Stimulation of phosphate uptake in human platelets by thrombin and collagen. Changes in specific 32P labeling of metabolic ATP and polyphosphoinositides. AB - The uptake of [32P]phosphate by human, gel-filtered blood platelets and its incorporation into cytoplasmic ATP and polyphosphoinositides was studied. In unstimulated platelets, uptake was Na+o-dependent and saturable at approximately 20 nmol/min/10(11) cells with a half-maximal rate at 0.5 mM extracellular phosphate. Upon stimulation with thrombin or collagen, net influx of [32P]Pi was accelerated 5- to 10-fold. With thrombin, [32P]Pi efflux was also increased. After the first 2 min, efflux exceeded influx, resulting in the net release of [32P]Pi from the platelets. Since the stimulus-induced burst in [32P]Pi uptake paralleled the secretory responses, it might be an integral part of stimulus response coupling in platelets. The stimulus-induced burst in net [32P]Pi uptake led to an enhanced labeling of metabolic ATP, which was already detectable at 5 s after stimulation with thrombin. Concomitantly, the incorporation of [32P]Pi into phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate was accelerated. The thrombin-induced increase in specific 32P radioactivity of cytoplasmic ATP fully accounted for the simultaneous increase in specific 32P radioactivity of these phosphoinositides. In studying the extent of 32P labeling of phosphorylated compounds in response to a cellular stimulus, it is therefore essential to measure the effect of the stimulus on the specific radioactivity of cytoplasmic ATP. PMID- 3034880 TI - Adenylyl cyclase in yeast. Hydrodynamic properties and activation by trypsin. AB - The adenylyl cyclase system of the yeast Saccharomyces cerevisiae contains the CYR1 polypeptide, responsible for catalyzing formation of cAMP from ATP, and two RAS polypeptides, responsible for stimulation of cAMP synthesis by guanine nucleotides. We have determined hydrodynamic properties of yeast adenylyl cyclase in taurocholate extracts of wild type and RAS-deficient membranes. In taurocholate extracts of both kinds of membranes, the enzyme is insensitive to guanine nucleotide stimulation; in the presence of 0.5 M NaCl, the taurocholate solubilized enzyme has a sedimentation coefficient of 12.5 S and a Stokes radius of 11 nm, consistent with a molecular weight of 594,000 for the protein-detergent complex. Treatment of particulate fractions with trypsin (less than 10 micrograms/ml) markedly activates membrane-bound adenylyl cyclase activity, abolishes stimulation by guanine nucleotides, and reduces the sedimentation coefficient of the detergent-solubilized enzyme; higher concentrations of trypsin release a still smaller water-soluble enzyme complex (7.5 S, 6.1 nm Stokes radius, calculated Mr = 190,000) from the membrane. In combination with genetic evidence (Kataoka, T., Broek, D., and Wigler M., (1985) Cell 43, 493-505), our data are consistent with a structural and functional model of yeast adenylyl cyclase in which GTP-activated RAS proteins stimulate cAMP synthesis by relieving an inhibitory constraint on the activity of the CYR1 gene product. This constraint may be mediated by the amino-terminal portion of the CYR1 polypeptide. PMID- 3034881 TI - Inhibition of Na+-H+ exchange by N,N'-dicyclohexylcarbodiimide in isolated rat renal brush border membrane vesicles. AB - The inactivation of rat renal brush border membrane Na+-H+ exchange by the covalent carboxylate reagent N,N'-dicyclohexylcarbodiimide (DCCD) was studied by measuring 1 mM Na+ influx in the presence of a pH gradient (pHi = 5.5; pHo = 7.5) and H+ influx in the presence of a Na+ or Li+ gradient ([Na+]i = 150 mM; [Na+]o = 1.5 mM). In the presence of DCCD, the rate of Na+ uptake decreased exponentially with time and transport inhibition was irreversible. At all DCCD concentrations the loss of activity was described by a single exponential, consistent with one critical DCCD-reactive residue within the Na+-H+ exchanger. Among several carbodiimides the most hydrophobic carbodiimide, DCCD, was also the most effective inhibitor of Na+-H+ exchange. With 40 nmol of DCCD/mg of protein, at 20 degrees C for 30 min, 75% of the amiloride-sensitive 1 mM Na+ uptake was inhibited. Neither the equilibrium Na+ content nor the amiloride-insensitive Na+ uptake was significantly altered by the treatment. The Na+-dependent H+ flux, measured by the change in acridine orange absorbance, was also decreased 80% by the same DCCD treatment. If 150 mM NaCl, 150 mM LiCl, or 1 mM amiloride was present during incubation of the brush border membranes with 40 nmol of DCCD/mg of protein, then Li+-dependent H+ flux was protected 50, 100, or 100%, respectively, compared to membranes treated with DCCD in the absence of Na+-H+ exchanger substrates. The combination of DCCD and an exogenous nucleophile, e.g. ethylenediamine and glycine methyl ester, increased Na+-dependent H+ flux in the presence of 80 nmol of DCCD/mg of protein, compared to the transport after DCCD treatment alone. These findings suggest that the Na+-H+ exchanger contains a single carboxylate residue in a hydrophobic region of the protein, and the carboxylate and/or a nearby endogenous nucleophilic group is critical for exchange activity. PMID- 3034883 TI - Biosynthesis of enterobacterial common antigen in Escherichia coli. In vitro synthesis of lipid-linked intermediates. AB - An in vitro system was developed to study the biosynthesis of enterobacterial common antigen (ECA). Membranes of Escherichia coli were found to possess an enzyme activity that catalyzes the transfer of UDP-N-acetyl-acetylglucosamine-1 phosphate from UDP-N-acetyl-glucosamine (UDP-GlcNAc) to an endogenous lipid acceptor according to the reaction UDP-GlcNAc + P-lipid----GlcNAc-PP-lipid + UMP. The lipid-linked product was tentatively identified as GlcNAc pyrophosphorylundecaprenol (lipid I) based on a comparison of its chemical and chromatographic properties with those of authentic GlcNAc pyrophosphorylundecaprenol. The enzyme was dependent on the presence of Mg2+ for activity, and the reaction catalyzed by the enzyme was totally inhibited by the antibiotic tunicamycin in both the forward and reverse directions. Incubation of membranes with both UDP-N-acetylmannosaminuronic acid (UDP-ManNAcA) and UDP GlcNAc resulted in the conversion of lipid I to a more polar compound, lipid II. The synthesis of lipid II was dependent on prior synthesis of lipid I. Characterization of the saccharide moiety of lipid II resulted in the identification of this compound as ManNAcA-GlcNAc-pyrophosphorylundecaprenol. PMID- 3034882 TI - Replacement of the invariant lysine 77 by arginine in yeast iso-1-cytochrome c results in enhanced and normal activities in vitro and in vivo. AB - Oligonucleotide-directed mutagenesis of the yeast Saccharomyces cerevisiae was used to generate an abnormal iso-1-cytochrome c having an Arg-77 replacement of the normal Lys-77; this Lys-77 residue is evolutionarily conserved in most eukaryotic cytochromes c and is trimethylated in fungal and plant cytochromes c. Examination of strains having a single chromosomal copy of the gene encoding the Arg-77 protein indicated that the altered protein was synthesized at the normal rate and that it had normal or near normal activity in vivo. Examination of enzymatic activities in vitro with cytochrome b2, cytochrome c peroxidase, and cytochrome c oxidase indicated that the altered iso-1-cytochrome c has equal or enhanced catalytic efficiencies. Thus, replacement of the evolutionarily conserved residue Lys-77 produces no or only minor effects both in vivo and in vitro. PMID- 3034884 TI - Metabolic pathways from 1 alpha,25-dihydroxyvitamin D3 to 1 alpha,25 dihydroxyvitamin D3-26,23-lactone. Stereo-retained and stereo-selective lactonization. AB - The present study was carried out in order to elucidate the metabolic pathway from 1 alpha,25-(OH)2D3 to 1 alpha,25-(OH)2D3-26,23-lactone. For that purpose, we stereospecifically synthesized the vitamin D3 derivatives 1 alpha,23(S),25 (OH)3D3, 1 alpha,23(S),25(R),26-tetrahydroxyvitamin D3, and 23(S),25(R)-1 alpha,25-dihydroxyvitamin D3-lactol. The in vitro metabolism of these compounds was examined in kidney homogenates and intestinal mucosa homogenates from 1 alpha,25-(OH)2D3-supplemented chicks. The naturally occurring 23(S),25(R)-1 alpha,25-dihydroxyvitamin D3-26,23-lactone was produced (in increasing amounts) from 1 alpha,25-(OH)2D3, 1 alpha,25(R),26-(OH)3D3, 1 alpha,23(S),25-(OH),D3, 1 alpha,23(S),25(R),26-(OH)4D3, and 23(S),25(R)-1 alpha,25-(OH)2D3-26,23-lactol. These results indicated that there are two possible metabolic pathways from 1 alpha,25-(OH)2D3 to 1 alpha,23(S),25(R),26-(OH)4D3: the major one is by way of 1 alpha,23(S),25-(OH)3D3 and the minor one is by way of 1 alpha,25(R),26-(OH)3D3. 1 alpha,23(S),25(R),26-Tetrahydroxyvitamin D3 is further metabolized to 23(S),25(R) 1 alpha,25-dihydroxyvitamin D3-26,23-lactone via 23(S),25(R)-1 alpha,25 dihydroxyvitamin D3-26,23-lactol. In the course of our studies, a new biosynthetic vitamin D3 metabolite was isolated in pure form. This metabolite was identified as 23(S),25(R)-1 alpha,25-(OH)2D3-26,23-lactol by UV spectrophotometry and mass spectrometry. Furthermore, we establish in this report that the lactonization of 1 alpha,23,25,26-(OH)4D3 and 1 alpha,25-(OH)2D3-26,23-lactol occurs in a stereo-retained and stereo-selective fashion. PMID- 3034885 TI - The role of oxygen radicals in dye-mediated photodynamic effects in Escherichia coli B. AB - Photosensitive dyes representative of the thiazines, xanthenes, acridines, and phenazines mediated phototoxicity in Escherichia coli B. The observed phototoxicity was sensitizer-, light-, and oxygen-dependent and is therefore a photodynamic effect. Hydroxyl radical scavengers conferred protection against the photodynamic action of all of the representative dyes. The extent of protection was dependent on the concentration of scavenger and on the in vitro reactivity of the scavenger with the hydroxyl radical. Exogenous superoxide dismutase and catalase partially protected the cells against the dye-mediated phototoxicity, and prior induction of intracellular superoxide dismutase and catalase by growth in glucose minimal medium containing manganese and paraquat substantially protected E. coli B against the photodynamic action of all of the dyes examined. Combinations of protective treatments against the phototoxicity of all four classes of dyes, including superoxide dismutase and catalase preinduction and addition of extracellular superoxide dismutase and catalase or the addition of hydroxyl radical scavengers, provided nearly complete protection against the oxygen-dependent component of dye-mediated lethality. E. coli B grown in glucose minimal medium containing manganese and photosensitive dyes induced manganese superoxide dismutase. The extent of induction was correlated with the dyes' ability to photooxidize NADH in vitro. Thus, oxygen radicals are primarily responsible for the oxygen-dependent toxicity of the photosensitive dyes examined, and one adaptive response of E. coli B to a dye-mediated oxidative threat is to induce superoxide dismutase. PMID- 3034886 TI - RNA polymerase pauses in vitro within the Escherichia coli origin of replication at the same sites where termination occurs in vivo. AB - An in vitro transcription system able to distinguish initiation at the 16-kDa promoter from elongation events was used to identify factors that might participate in transcription termination within oriC. Pausing in the oriC region occurs at the same sites where termination occurs in vivo. Ten of these sites overlap RNA:DNA junctions in oriC. The pausing that occurs in vitro was not converted to efficient termination by guanosine 5'-diphosphate 3'-diphosphate, NusA, and Rho alone or in combination, or by DnaA suggesting that in vivo other or additional factors contribute to termination at oriC. Transcription from the 16-kDa promoter was 90% inhibited by the nucleotides guanosine 5'-diphosphate 3' diphosphate and guanosine 5'-triphosphate 3'-diphosphate in agreement with previous observations that this promoter is stringently regulated. PMID- 3034887 TI - Synthesis and regulation of the two human complement C4 genes in stable transfected mouse fibroblasts. AB - The major histocompatibility complex-linked human complement C4 genes are highly homologous in primary structure but give rise to products which differ in complement-activating function. In order to examine the synthesis, function, and regulation of these two genes independently, cloned C4A and C4B genes were transfected into mouse fibroblast L-cells. In the stable transfected cell lines, C4A and C4B are synthesized, undergo a complex series of post-translational modifications, and each functions appropriately in activation of the classical complement pathway. A marked difference in the kinetics of complement component C1-mediated cleavage of the C4A- and C4B-alpha chains was demonstrated in the transfectants and may contribute to the differences in the intrinsic functional activity of the two C4 isotypes. In contrast to the expression of other complement genes which are affected during the hepatic "acute phase response" (factor B, C3), the expression of C4 was not regulated by interleukin-1 or tumor necrosis factor. Interferon-gamma, however, mediated a dose- and time-dependent increase in the expression of the C4 genes. Moreover, interferon had a significantly greater and longer-lasting effect on the synthesis of C4A than that of C4B. Differences in the expression and regulation of these two genes provide insight into the control of complement activation during inflammation. PMID- 3034888 TI - Hormonal regulation of the synthesis and mRNA content of the regulatory subunit of cyclic AMP-dependent protein kinase type II in cultured rat ovarian granulosa cells. AB - These studies were undertaken to determine the molecular events by which estradiol and follicle-stimulating hormone (FSH) stimulate in ovarian granulosa cells the increase in the content of one of the regulatory subunits of cAMP dependent protein kinase type II, RII51 (Mr = 51,000), and its electrophoretic variants RII51.5 (Mr = 51,500) and RII52 (Mr = 52,000). To analyze the de novo synthesis of RII51/52, granulosa cells were cultured (10(6) cells/ml) for 0, 12, 24, or 48 h with estradiol (10 nM) +/- FSH (12.5, 25, and 50 ng/ml), 8-bromo-cAMP (0.25-3 mM), or forskolin (0.5-100 microM) and then pulse-labeled with [35S]methionine (300 microCi/ml; 4 h). Labeled RII51, present either in urea extracts of total cellular protein or after partial purification from a soluble cell extract by cAMP-Sepharose chromatography, was quantitated by autoradiography of two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels and by excision of the silver-stained spots of the RII51 variants from the gels and counting. Synthesis of RII51 and its electrophoretic variants was low in cells cultured with estradiol alone for 48 h, whereas it was increased 4-5-fold in cells cultured with estradiol and FSH. Changes in the synthesis of actin were minor throughout the culture period regardless of hormone treatment. Pulse-chase experiments using [35S]methionine provided evidence that the isoelectric variants RII51.5 and RII52 may be derived from RII51 by post-translation modification, such as phosphorylation. Labelling with [32P]orthophosphate showed that RII52 contained more radioactivity than RII51.5 and RII51. Northern and filter hybridization assays demonstrated a 6-10-fold dose- and time-dependent increase in the amount of RII51 mRNA in granulosa cells exposed to estradiol and FSH or estradiol and forskolin compared to those cultured with estradiol alone. In vitro translation of poly(A)+ mRNA of granulosa cells from estradiol- and FSH-treated hypophysectomized rats also demonstrated an increase in the content of translatable RII51 mRNA. These studies indicate that in cultured rat granulosa cells the synthesis of RII51 and the content of its mRNA are selectively increased by estradiol and cAMP in a time- and dose-dependent manner. Based on these observations, RII51 appears to be a useful marker to determine the molecular (genomic?) sites of estradiol and FSH action in differentiating rat granulosa cells. PMID- 3034889 TI - Delineation of the intronless nature of the genes for the human and hamster beta 2-adrenergic receptor and their putative promoter regions. AB - The beta 2-adrenergic receptor is the first adenylate cyclase-coupled receptor to be cloned. We provide here a detailed characterization of its complete gene in both the human and hamster which reveals several unusual and provocative features. The genes are present in a single copy, are intronless, and are bounded by homologous 18-bp (base pair) direct repeats. These findings suggest that the beta 2-adrenergic receptor may have arisen as a processed gene for another related gene. Genomic Southern blots done at reduced stringency in fact reveal additional weak signals. The human and hamster gene sequences 5' to the principal site of transcription initiation are highly homologous and share many characteristics of promoters for housekeeping genes. Moreover, there is present in the human genome a long (777 bp) open reading frame which is in frame with the beta-adrenergic receptor coding block and which ends only 234 bp 5' to the initiator methionine of the receptor. An unusual cDNA has been found, transcribed from a putative second more 5' promoter which contains the 5' half of the beta adrenergic receptor as well as 1065-bp 5' to the receptor coding region, including the entire upstream long open reading frame (sufficient to encode a putative protein of Mr approximately 28,000). PMID- 3034892 TI - The human pyruvate dehydrogenase complex. Isolation of cDNA clones for the E1 alpha subunit, sequence analysis, and characterization of the mRNA. AB - cDNA clones corresponding to the entire length of mRNA for the alpha subunit of human pyruvate dehydrogenase (EC 1.2.4.1), the E1 component of the pyruvate dehydrogenase complex, have been isolated from liver cDNA libraries. Two classes of cDNA clones were obtained and these correspond to two forms of pyruvate dehydrogenase E1 alpha mRNA. Both mRNA species have been demonstrated in a variety of human tissues and cultured fibroblasts. The cDNA sequence has been determined and, from it, the protein sequence of the human E1 alpha subunit was deduced. The protein is synthesized with a typical mitochondrial import leader sequence and the peptide bond at which this sequence is cleaved after transport into the mitochondrion has been determined by direct amino acid sequencing of the mature E1 alpha subunit. The human pyruvate dehydrogenase E1 alpha subunit contains identical phosphorylation sites to those found in the corresponding porcine protein. Preliminary studies of pyruvate dehydrogenase E1 alpha mRNA in cultured fibroblasts from patients with severe pyruvate dehydrogenase deficiency have revealed considerable heterogeneity as would be expected from protein studies. PMID- 3034890 TI - Adenovirus late protein synthesis is resistant to the inhibition of translation induced by poliovirus. AB - Inhibition of host protein synthesis after poliovirus infection has been suggested to be a consequence of the proteolytic degradation of a p220 polypeptide necessary to translate capped mRNAs. However, the synthesis of several adenovirus late proteins on capped mRNAs was resistant to poliovirus inhibition. Thus, the hexon protein was still made 8 h after poliovirus superinfection. The synthesis of other adenovirus proteins such as the fiber was much more sensitive to poliovirus-induced inhibition than the hexon, either in the absence or in the presence of guanidine. Detailed densitometric analyses clearly showed the differential behavior of several adenovirus late mRNAs to poliovirus shut-off of translation. This is striking in view of the fact that a common leader sequence in the 5' termini is present in the adenovirus late mRNAs. The use of 3-methyl quercetin, an inhibitor of poliovirus RNA synthesis (Castrillo, J. L., Vanden Berghe, D., and Carrasco, L. (1986) Virology 152, 219 227), showed that translation of several capped adenovirus mRNAs took place in poliovirus-infected cells after the synthesis of host proteins had ceased. The poliovirus mRNA and the adenovirus mRNA coding for the hexon protein are very efficient mRNAs and have a leader sequence of more than 740 and 250 nucleotides, respectively, with very rich secondary structures making it difficult to predict how the scanning model will operate on these two mRNAs. PMID- 3034891 TI - Co-purification and characterization of ATP-sulfurylase and adenosine-5' phosphosulfate kinase from rat chondrosarcoma. AB - The two sulfate-activating enzymes, ATP-sulfurylase (EC 2.7.7.4) and adenosine-5' phosphosulfate kinase (adenylylsulfate kinase, EC 2.7.1.25), were each purified about 2000-fold from crude rat chondrosarcoma homogenate. Throughout a purification protocol which included Sephacryl S-300 gel filtration, DEAE Sephadex ion exchange, hydroxylapatite, and ATP-agarose affinity chromatography, these two activities consistently co-purified. ATP-sulfurylase and adenosine-5' phosphosulfate kinase each showed a pH optima of 7.0-7.4 and a bimodal temperature optima of 46 and 52-54 degrees C. Both activities preferred Mg2+ as their divalent cation source over Mn2+, Co2+, or Zn2+. The apparent Km values determined for adenosine 5'-phosphosulfate in both assays was 1-5 microM; the Km for pyrophosphate in the sulfurylase reaction was 40 microM and for ATP in the kinase reaction was 5 mM. Gel electrophoresis indicated major bands at Mr = 160,000 in nondenaturing systems and 35,000-37,000 and 60,000 under dissociative conditions, whereas gel filtration of the most highly purified fractions yielded a coincident peak in the molecular weight range 260,000. PMID- 3034893 TI - Complete purification and general characterization of FAD synthetase from rat liver. AB - Flavin adenine dinucleotide synthetase (ATP:FMN adenylyltransferase, EC 2.7.7.2) was purified about 10,000-fold from the high-speed supernatant of rat liver by a sequence of ammonium sulfate fractionation and column chromatographies on DEAE Sephadex (A-50), chromatofocusing, FMN-agarose affinity, and Sephadex G-200. The specific activity of the purified enzyme was 133 units (nanomoles of FAD formed per min at 37 degrees C)/mg of protein. This preparation was free from contaminating FAD pyrophosphatase. The apparent molecular weight was estimated to be 97,000 by gel filtration on Sephadex G-200. Sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed an apparent subunit molecular weight of 53,000. Hence, the enzyme is a dimer of approximately 100,000. The enzyme was found most active at pH 7.1, requires Mg2+, and is essentially irreversible in the direction of FAD formation. Kinetic analysis gave Km values of 9.6 microM for FMN and 53 microM for ATP. PMID- 3034894 TI - Identification and characterization of a high density lipoprotein-binding protein in cell membranes by ligand blotting. AB - Cholesterol efflux from cultured cells can be mediated through binding of high density lipoprotein (HDL) to a cell-surface site which shows many characteristics of a biological receptor. To determine whether a specific protein forms a component of this site, cell membrane proteins were analyzed by ligand blotting using 125I-HDL3. Results demonstrated that membranes from a number of cell types possess a protein with an apparent molecular mass of 110 kDa that binds HDL and apoA-I and apoA-II proteoliposomes, but not low density lipoprotein, acetylated low density lipoprotein, or apoE proteoliposomes. The binding activity of this protein was increased by loading cells with cholesterol and was abolished by trypsin treatment of intact cell monolayers. These results suggest that HDL binds with specificity to a cell-surface protein which is regulated by intracellular cholesterol levels. Since HDL binding to intact cell monolayers shows the same characteristics, the 110-kDa binding protein may represent the proposed HDL receptor that functions to facilitate transport of cholesterol from cells to HDL particles. PMID- 3034895 TI - Isolation and properties of a mutant of Escherichia coli possessing defective Na+/H+ antiporter. AB - A mutant of Escherichia coli with defective Na+/H+ antiporter was isolated. The rationale for its isolation was that cells possessing defective Na+/H+ antiporter, which is essential for establishment of a Na+ gradient, could not grow with a carbon source that was taken up with Na+. The mutant had no appreciable Na+/H+ antiporter activity, but its K+/H+ antiporter and Ca2+/H+ antiporter activities were normal. Judging from the reversion frequency, the defect seems to be due to a single mutation. The mutant could not grow at alkaline pH. Therefore, the Na+/H+ antiporter, but not the K+/H+ antiporter or the Ca2+/H+ antiporter, seems to be responsible for pH regulation in alkaline medium. This mutant will be useful for cloning the Na+/H+ antiporter gene and for detection of Na+-substrate cotransport systems. PMID- 3034896 TI - A cholera toxin substrate regulates cyclic GMP content of rat pinealocytes. AB - The adrenergic regulation of cyclic GMP in isolated pinealocytes was investigated. In this cell, norepinephrine stimulates cyclic GMP and cyclic AMP greater than 100-fold by activating both alpha 1- and beta-adrenoceptors. beta Adrenergic activation is a requisite event and is potentiated by alpha 1 adrenergic activation (Vanecek, J., Sugden, D., Weller, J. L., and Klein, D. C. (1985) Endocrinology 116, 2167-2173). The current study found that cholera toxin could substitute for beta-adrenergic agonists in stimulating pinealocyte cyclic GMP content, as has been found to be the case for cyclic AMP. Treatment with cholera toxin alone (1 microgram/ml for 90 min) had a small effect (2- to 4-fold increase) on cyclic GMP; addition of the alpha 1-adrenergic agonists, phenylephrine, cirazoline, or methoxamine to cholera toxin-treated cells rapidly (peak at 5 min) caused a further 30- to 300-fold increase. The alpha 1-adrenergic agonists had little effect by themselves at concentrations which potentiated the effects of cholera toxin. The potentiating effect of phenylephrine was inhibited nearly completely by an alpha 1-adrenergic antagonist, but not by either an alpha 2- or beta-adrenergic antagonist. The purified cholera toxin subunits A and B did not stimulate cyclic GMP either alone or in the presence of phenylephrine. Furthermore, the potentiating action of phenylephrine was observed following 90 min but not 20 min of cholera toxin pretreatment. these results suggest that the regulation of cyclic GMP levels in the pineal gland involves an Ns-like GTP binding regulatory protein. This is of interest because it is the first indication that cyclic GMP is regulated by such a GTP-binding protein in nonretinal tissue. It remains to be determined whether the mechanisms involved in the transmembrane regulation of cyclic AMP and cyclic GMP in any other tissue are similar. PMID- 3034897 TI - Orangutan fetal globin genes. Nucleotide sequence reveal multiple gene conversions during hominid phylogeny. AB - We have determined the nucleotide sequences of the linked gamma 1- and gamma 2- fetal globin genes from a single orangutan (Pongo pygmaeus) chromosome and compared them with the corresponding genes of other simian primates (gamma 1- and gamma 2-genes of human, chimpanzee, gorilla, and the single gamma-gene of the spider monkey). Previous studies have indicated that the two gamma-gene loci in catarrhine primates resulted from a duplication about 25-35 million years ago. However, comparisons of aligned gamma-gene sequences show that these genes contain three regions with distinct histories of which only the 3' third clearly reflects the ancestral nature expected of the gamma-gene duplication. To explain these different evolutionary histories and also hominid relationships we provide evidence for the occurrence of sequence conversions which affect region 1 (120 base pairs 5'-flanking through exon 2) in all hominid species and extend to varying degrees into region 2 (intron 2 through exon 3). Close examinations of the proposed conversions further suggest that 12 of the 13 conversions identified involved gamma 1 converting gamma 2. Polarity of these conversions may be a result of differential survival between these genes because during human fetal development the gamma 1-gene is preferentially expressed over the gamma 2-gene and it may be subjected to greater selection pressure to remain unaltered. PMID- 3034898 TI - Distinct steps in the import of ADP/ATP carrier into mitochondria. AB - Transport of the precursor to the ADP/ATP carrier from the cytosol into the mitochondrial inner membrane was resolved into several consecutive steps. The precursor protein was trapped at distinct stages of the import pathway and subsequently chased to the mature form. In a first reaction, the precursor interacts with a protease-sensitive component on the mitochondrial surface. It then reaches intermediate sites in the outer membrane which are saturable and where it is protected against proteases. This translocation intermediate can be extracted at alkaline pH. We suggest that it is anchored to the membrane by a so far unknown proteinaceous component. The membrane potential delta psi-dependent entrance of the ADP/ATP carrier into the inner membrane takes place at contact sites between outer and inner membranes. Completion of translocation into the inner membrane can occur in the absence of delta psi. A cytosolic component which is present in reticulocyte lysate and which interacts with isolated mitochondria is required for the specific binding of the precursor to mitochondria. PMID- 3034899 TI - Complete amino acid sequence of the collagenase from the insect Hypoderma lineatum. AB - The primary structure of the Hypoderma lineatum collagenase was determined. Chymotrypsin digestion and thermolysin fragmentation of the chymotryptic core gave 30 and 5 peptides, respectively, accounting for all the residues of the protein. These peptides were aligned with overlapping peptides derived from tryptic and Staphylococcus aureus V8 proteinase digests. Hypoderma collagenase is a serine proteinase composed of 230 amino acids (Mr 25,223). It displays a high degree of sequential homology with the serine proteinases of the trypsin family, especially with another collagenolytic enzyme, the proteinase I of the crab Uca pugilator. The six half-cystinyl residues of Hypoderma collagenase correspond to 6 of the 10 half-cystinyl residues of chymotrypsin, and the residues forming the charge-relay system of the active site of chymotrypsin (His-57, Asp-102, and Ser 195) are found in corresponding regions. The prediction of the secondary structure of the collagenase is given. PMID- 3034901 TI - Expression of human angiotensinogen cDNA in Escherichia coli. AB - Human angiotensinogen cDNA clones were isolated from a human liver library. Nucleotide sequence analysis of these cDNA clones revealed that position 1075 in the messenger RNA, which is part of a PstI recognition sequence, is different from the published sequence (Kageyama, R., Ohkubo, H., and Nakanishi, S. (1984) Biochemistry 23, 3603-3609). This change results in an altered amino acid at this position in the corresponding protein sequence and suggests possible restriction fragment length polymorphism. The full length human angiotensinogen cDNA was constructed from partial cDNA clones and ligated into an isopropyl-1-thio-beta-D galactopyranoside inducible bacterial expression vector pUC9 to develop expression plasmid pUCHAG27. This plasmid permitted the synthesis of human angiotensinogen in Escherichia coli. The recombinant bacteria overproduced a 53 kDa protein which was recognized by anti-human angiotensinogen antibodies. The synthesis of this protein was greatly increased upon induction with isopropyl-1 thio-beta-D-galactopyranoside. The chimeric protein, almost identical to human angiotensinogen, was partially purified by ammonium sulfate fractionation and gel filtration on Sephadex G-100. Human kidney renin was shown to enzymatically cleave this recombinant protein to produce des-(angiotensin I)-angiotensinogen and a small polypeptide. Thus, we provide evidence that recombinant human angiotensinogen synthesized through E. coli is biologically active and serves as a substrate for human renin. PMID- 3034902 TI - Down-regulation of cell surface cyclic AMP receptors and desensitization of cyclic AMP-stimulated adenylate cyclase by cyclic AMP in Dictyostelium discoideum. Kinetics and concentration dependence. AB - cAMP binds to Dictyostelium discoideum surface receptors and induces a transient activation of adenylatecyclase, which is followed by desensitization. cAMP also induces a loss of detectable surface receptors (down-regulation). Cells were incubated with constant cAMP concentrations, washed free of cAMP, and cAMP binding to surface receptors and cAMP-induced activation of adenylate cyclase were measured. cAMP could induce maximally 65% loss of binding activity and complete desensitization of cAMP-stimulated adenylate cyclase activity. Half maximal effects for down-regulation were observed at 50 nM cAMP and for desensitization at 5 nM cAMP. Down-regulation was rapid with half-times of 4, 2.5, and 1 min at 0.1, 1, and 10 microM cAMP, respectively. Similar kinetic data have been reported for desensitization (Dinauer, M.C., Steck, T.L., and Devreotes, P.N. (1980) J. Cell Biol. 86, 554-561). Down-regulation and desensitization were not reversible at 0 degrees C. Down-regulation reversed slowly at 20 degrees C with a half-time of about 1 h. Resensitization of adenylate cyclase was biphasic showing half-times of 4 min and about 1 h, respectively; the contribution of the rapidly resensitizing component was diminished when down-regulation of receptors was enhanced. These results suggest that cAMP-induced down-regulation of receptors and desensitization of adenylate cyclase stimulation proceed by at least two steps. One step is rapidly reversible, occurs at low cAMP concentrations, and induces desensitization without down-regulation, while the second step is slowly reversible, requires higher cAMP concentrations, and also induces down-regulation. PMID- 3034900 TI - Genetic factors controlling structure and expression of apolipoproteins B and E in mice. AB - We report the identification and partial characterization of polymorphisms among inbred strains of mice affecting several aspects of the expression of apolipoproteins B and E (apoB and apoE), the major proteins of low density lipoproteins (LDL) and very low density lipoproteins (VLDL). These polymorphisms include differences in the levels of the lipoproteins and apolipoproteins on both chow and high fat diets, differences in their response to a high fat diet challenge, and differences in the relative levels of the two molecular weight species of apoB. Although most strains exhibited a large increase in plasma LDL and VLDL in response to a high fat diet, the levels of apoB and apoE mRNA were either unaffected or, in some cases, decreased slightly. Also, the levels of apoB and apoE mRNA were not correlated among strains with the levels of the apolipoproteins in plasma, suggesting that genetic control occurs primarily at the level of lipoprotein catabolism. Elucidation of the precise mechanisms involved in the differences will require genetic analysis. Toward this end, we have identified DNA polymorphisms for apoB and apoE and have used these in segregation analysis to determine the chromosomal locations of the apoB and apoE structural genes in mice. The gene for apoB, designated Apob, resides in the proximal region of chromosome 12 linked to genes for ribosomal RNA and aryl hydrocarbon hydroxylase. The gene for apoE, designated Apoe, is located on chromosome 7, linked to genes for glucose phosphate isomerase and peptidase 4. Previously, we mapped the structural genes for apolipoproteins A-I and A-II to mouse chromosomes 9 and 1, respectively, and thus, the four loci encoding mammalian apolipoproteins have now been located in the mouse. These loci are homologous to the loci encoding apolipoproteins in humans as judged by the conservation of linked markers. A correlation was observed between a unique apoB allele and "responsiveness" to a high fat diet challenge. There were no obvious associations of apoB, apoE, or LDL/VLDL phenotypes or genotypes with diet-induced atherosclerosis among strains surveyed. These results clarify the organization and regulation of the genes for apoB and apoE, and they provide information about the naturally occurring polymorphisms affecting their expression. PMID- 3034903 TI - The peroxide complex of yeast cytochrome c peroxidase contains two distinct radical species, neither of which resides at methionine 172 or tryptophan 51. AB - The nature of the free radical species observed in the peroxide complex of yeast cytochrome c peroxidase is described for protein variants containing amino acid substitutions at Met-172 and Trp-51. As was the case with Met-172 mutations (Goodin, D.B., Mauk, A.G., and Smith, M. (1986) Proc. Natl. Acad. Sci. U.S.A. 83, 1295-1299), Trp-51 can be substituted to give active enzyme. Phe-51-containing enzyme has a higher turnover rate than the original enzyme and exhibits an altered pH dependence. The properties of the isotropic and axial components (Hoffman, B.M., Roberts, J.E., Kang, C.H., and Margoliash, E. (1981) J. Biol. Chem. 256, 6556-6564; Hori, H., and Yonetani, T. (1985) J. Biol. Chem. 260, 349 355) of the EPR signal of the wild-type enzyme-peroxide complex, studied as a function of H2O2 stoichiometry, support proposals (Goodin et al. (1985) and Hori and Yonetani (1985), see above) that two distinct radical species are formed, and spin quantification shows that the isotropic radical is always formed in substoichiometric amounts. The peroxide complexes for proteins containing amino acid substitutions at either Met-172 or Trp-51 exhibit somewhat larger than normal levels of the isotropic radical signal. In addition, these mutants are unlike wild-type enzyme in that the axial EPR signal associated with the peroxide complex is seen only at 10 K and not at 90 K. Thus, neither amino acid can be considered to be the molecular species responsible for either radical signal, but both mutations appear to affect the physical properties of the axial signal representing the major radical species. PMID- 3034904 TI - Polyphosphoinositide labeling in rat liver plasma membranes is reduced by preincubation with cholera toxin. AB - Incubation of a crude rat liver plasma membrane preparation with [gamma-32P]ATP resulted in a rapid Mg2+-dependent incorporation of 32P into phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate. Preincubation of the membranes with cholera toxin under ADP-ribosylating conditions reduced the labeling of the polyphosphoinositides. This action of cholera toxin required NAD+ and guanine nucleotides, was dose-dependent with respect to cholera toxin, and could not be mimicked by cAMP. It therefore appears that ADP-ribosylation of the stimulatory guanine nucleotide-binding regulatory protein of adenylate cyclase, or another G-protein, in rat liver plasma membranes affects the activity of enzymes in the polyphosphoinositide pathway. PMID- 3034905 TI - Topological analysis of the major protein in isolated intact rat liver gap junctions and gap junction-derived single membrane structures. AB - The topological organization of the major rat liver gap junction protein has been examined in intact gap junctions and gap junction-derived single membrane structures. Two methods, low pH and urea at alkaline pH, were used to "transform" or "split" double membrane gap junctions into single membrane structures. Low pH treatment "transforms" rat liver gap junctions into small single membrane vesicles which have an altered sodium dodecyl sulfate-polyacrylamide gel electrophoresis profile after digestion with L-1-to-sylamido-2 phenylethylchloromethyl ketone-trypsin. Alkaline pH treatment in the presence of 8 M urea can split isolated rat liver gap junctions into single membrane sheets which have no detectable structural alteration or altered sodium dodecyl sulfate polyacrylamide gel electrophoresis profile after proteolytic digestion, suggesting that these single membrane sheets may be useful for topological studies of the gap junction protein. Proteolytic digestion studies have been used to localize the carboxyl terminus of the molecule on the cytoplasmic surface of the intact gap junction. However, the amino terminus does not appear to be accessible to proteases or to interaction with an antibody that is specific for the amino-terminal region of the molecule in intact or split gap junctions. Binding of antibodies, that block junctional channel conductance, can be eliminated by proteolytic digestion of intact gap junctions, suggesting that all antigenic sites for these antibodies are located on the cytoplasmic surface of the intact gap junction. In addition, calmodulin gel overlays indicate that at least two calmodulin binding sites exist on the cytoplasmic surface of the junctional protein. The information generated from these studies has been used to develop a low resolution two-dimensional model for the organization of the major rat liver gap junctional protein in the junctional membrane. PMID- 3034907 TI - Regulation of dnaB function in DNA replication in Escherichia coli by dnaC and lambda P gene products. AB - The dnaB protein of Escherichia coli, a multifunctional DNA-dependent ribonucleotide triphosphatase and dATPase, cross-links to ATP on ultraviolet irradiation under conditions that support rNTPase and dATPase activities of dnaB protein. The covalent cross-linking to ATP is specifically inhibited by ribonucleotides and dATP. Tryptic peptide mapping demonstrates that ATP cross links to only the 33-kDa tryptic fragment (Fragment II) of dnaB protein. The presence of single-stranded DNA alters the covalent labeling of dnaB protein by ATP, suggesting a possible role of DNA on the mode of nucleotide binding by dnaB protein. Present studies demonstrate that the dnaC gene product binds ribonucleotides independent of dnaB protein. On dnaB-dnaC protein complex formation, covalent incorporation of ATP to dnaB protein decreases approximately 70% with a concomitant increase of ATP incorporation to dnaC protein by approximately 3-fold. The mechanism of this phenomenon has been analyzed in detail by titrating dnaB protein with increasing amounts of dnaC protein. The binding of dnaC protein to dnaB protein appears to be a noncooperative process. The lambda P protein, which interacts with dnaB protein in the bacteriophage lambda DNA replication, does not bind ATP in the presence or absence of dnaB protein. However, lambda P protein enhances the covalent incorporation of ATP to dnaB protein approximately 4-fold, suggesting a direct physical interaction between lambda P and dnaB proteins with a probable change in the modes of nucleotide binding to dnaB protein. The lambda P protein likely forms a lambda P dnaB-ATP dead-end ternary complex. The implications of these results in the E. coli and bacteriophage lambda chromosomal DNA replication are discussed. PMID- 3034906 TI - Isolation of the yeast gene encoding elongation factor 3 for protein synthesis. AB - The gene YEF-3 encoding the elongation factor 3 (EF-3) for peptide chain elongation in Saccharomyces cerevisiae has been isolated by immunoscreening of a yeast genomic library in the phage lambda gt11. The identity of the EF-3 gene was confirmed by several methods. First, a clone-encoded protein could affinity purify the antibody that specifically reacted with EF-3. Second, a recombinant fusion protein, which reacted with anti-beta-galactosidase antibody as well as with anti-EF-3 antibody, was found in the lysate of a positive clone lysogen. Third, the function of EF-3 in a yeast mutant in which the EF-3 activity is temperature-sensitive in an in vitro assay could be complemented by transformations. EF-3 protein was overproduced in a transformant which contained the EF-3 gene on a multicopy plasmid YEp-13. Southern blot analysis shows that YEF-3 is a single copy gene. The transcript unit as mapped by S1 nuclease mapping, is consistent with the size of the message determined by Northern blot analysis and shows no evidence of introns. PMID- 3034908 TI - Identification of the multidrug resistance-related membrane glycoprotein as an acceptor for calcium channel blockers. AB - A radioactive photoactive dihydropyridine calcium channel blocker, [3H]azidopine, was used to photoaffinity label plasma membranes of multidrug-resistant Chinese hamster lung cells selected for resistance to vincristine (DC-3F/VCRd-5L) or actinomycin D (DC-3F/ADX). Sodium dodecyl sulfate-polyacrylamide gel electrophoretic fluorograms revealed the presence of an intensely radiolabeled 150-180-kDa doublet in the membranes from drug-resistant but not from the drug sensitive parental (DC-3F) cells. A similar radiolabeled doublet was barely detected in a drug-sensitive partial revertant (DC-3F/ADX-U) cell line. The 150 180-kDa doublet exhibited a specific half-maximal saturable photolabeling at 1.07 X 10(-7) M [3H]azidopine. The dihydropyridine binding specificity was established by competitive blocking of specific photolabeling with nonradioactive azidopine as well as with nonphotoactive calcium channel blockers nimodipine, nitrendipine, and nifedipine. In addition, [3H]azidopine photolabeling was blocked by verapamil and diltiazem but was stimulated by excess prenylamine and bepridil suggesting a cross-specificity for up to four different classes of calcium channel blockers. The 150-180-kDa calcium channel blocker acceptor co-electrophoresed exactly with the 150-180-kDa surface membrane glycoprotein (gp150-180 or P-glycoprotein) Vinca alkaloid acceptor from multidrug-resistant cells and was immunoprecipitated by polyclonal antibody recognizing gp150-180. [3H]Azidopine photolabeling of the 150 180-kDa component in the presence of excess vinblastine was reduced over 90%, confirming the identity or close relationship of the calcium channel blocker acceptor and the gp150-180 Vinca alkaloid acceptor. The [3H]azidopine photolabeling of gp150-180 also was reduced by excess actinomycin D, adriamycin, or colchicine, demonstrating a broad gp150-180 drug recognition capacity. The ability of gp150-180 to recognize multiple natural product cytotoxic drugs as well as calcium channel blockers suggests a direct function for gp150-180 in the multidrug resistance phenomenon and a role in the circumvention of that resistance by calcium channel blockers. PMID- 3034910 TI - Processing of the platelet-derived growth factor receptor. Biosynthetic and degradation studies using anti-receptor antibodies. AB - Studies of platelet-derived growth factor (PDGF) receptor biosynthesis and degradation have been limited by the lack of anti-receptor antibodies. In this study, peptides based on the cDNA-predicted amino acid sequence of the PDGF receptor were used to produce antisera that specifically immunoprecipitated the receptor. PDGF receptor biosynthesis was examined by pulse-chase labeling of cultured fibroblasts with [35S]methionine followed by immunoprecipitation. In BALB/c 3T3 fibroblasts the receptor was synthesized as a 160-kDa precursor that was converted to a mature 180-kDa form within 30-45 min. Removal of high mannose oligosaccharides by endo-beta-N-acetylglucosaminidase H treatment reduced the apparent molecular weight of the 160-kDa precursor but did not affect the migration of the 180-kDa mature receptor. When mannosidase II was inhibited by swainsonine, the 160-kDa precursor failed to mature; instead a 168-kDa form of the receptor was observed. Nevertheless, swainsonine-treated cells responded mitogenically to PDGF. The mature 180-kDa form of the receptor had a half-life of approximately 3 h in the absence of ligand. Addition of PDGF reduced the receptor half-life to 45 min. These studies define and characterize a PDGF receptor precursor, show that receptor degradation is enhanced by PDGF, and demonstrate the functional integrity of incompletely processed PDGF receptors. PMID- 3034909 TI - Mosaic evolution of the insulin-like growth factors. Organization, sequence, and expression of the rat insulin-like growth factor I gene. AB - Insulin-like growth factor I (IGF-I) plays a major role in mammalian growth and regenerative processes as a mediator of many of the biological effects of growth hormone. We have demonstrated recently that the human IGF-I gene is transcribed and processed into distinct messenger RNA molecules, each of which directs the synthesis of unique IGF-I-containing peptides. As a means to determine whether a similar model of IGF-I gene organization and expression is the paradigm in mammals and as an initial step in devising experimental approaches to the study of regulation of IGF-I biogenesis, we have isolated and characterized the rat IGF I gene. The rat gene, like its human counterpart, is very large, extending over at least 73 kilobases, and is composed of five exons subdivided by four introns. As in the human example, the rat IGF-I gene hybridizes to several messenger RNAs: 0.8-1.2, 1.6-2.1, and 7.8 kilobases. There is extensive nucleotide and amino acid sequence conservation between the two genes. The predicted mature rat IGF-I protein is identical to the human peptide in 67 of 70 residues. A comparably high degree of amino acid sequence identity is also found for both the amino- and carboxyl-terminal extension peptides, suggesting that, like mature IGF-I, the extension molecules may have physiological function. PMID- 3034911 TI - Aging test and dynamic fatigue test of apatite-wollastonite-containing glass ceramics and dense hydroxyapatite. AB - The purpose of this study is to examine the changes in mechanical strength of two bioactive ceramics in living tissue. An aging test and dynamic fatigue test were performed using apatite-wollastonite-containing glass ceramics (A X W-GC) and dense hydroxyapatite (HA). Specimens (5 mm X 5 mm X 25 mm, abraded with No. 2000 Al2O3 powder) were implanted into subcutaneous tissue of rats for varying periods of time. The bending strength of aged samples was measured by the three-point loading method. The bending strength of A X W-GC was greater than that of HA (P less than 0.001). There was no reduction in bending strength for both A X W-GC and HA in living tissue. The n value of both A X W-GC and HA did not decrease significantly after implantation as assessed by the results of the dynamic fatigue test according to analysis of covariance. SEM-EPMA showed that Si and Mg contents decreased, Ca content did not change, while P content increased in the surface of A X W-GC. The area where x-ray intensity changed increased moderately after implantation. There were no changes in Ca and P at the interface between HA and soft tissue. In macroscopic and microscopic observations, specimens were found to be encapsulated with a thin layer of connective tissue. Foreign body giant cells, osteoblasts, or osteoclasts were not observed in the soft tissue. There was no bonding between ceramics and soft tissue. PMID- 3034912 TI - A histological comparison of the tissue interface of bioglass and silica glass. AB - The objectives of the present article are to confirm the bone bonding phenomenon of Bioglass (BG) developed by Hench et al., and to observe the singularity of tissue reaction to it. BG and nonreactive silica glass (SG) were implanted in the femurs of rabbits and rats. Histological examination revealed that a relatively acellular zone with little inflammation was formed on BG surface at 1 day after implantation. Neither fibrous tissue nor a distinct boundary was observed between BG and bone after 7 days. On the contrary, a moderate postoperative inflammatory reaction was observed on SG at 1 day, and fibrous tissue was observed between SG and bone after 7 days. From these findings, it was confirmed that BG bonded directly with bone. As the relatively acellular zone observed on BG surface at 1 day was replaced by bone after 7 days, the formation of this zone might play an important role in bone-bonding process. Further research should be focused on the mechanism and biological meaning of bone bonding, for this phenomenon can not be explained by the conventional pathological theory of foreign body encapsulation. PMID- 3034913 TI - Reversible morphological and functional abnormalities of RINm5F cells cultured on polystyrene sulfonate beads. AB - RINm5F cells (an insulin-secreting cell line) were cultured on PSSO3Na microbeads under static conditions. The cell growth rate was either identical to that of cells grown on plastic wells or slower, depending on the initial cell concentration. With both supports, it was similarly influenced by the fetal calf serum concentration in the culture medium, and protein content per cell was identical. However, no spreading was observed when cells were cultured on microbeads. RINm5F cells cultured on plastic wells responded to arginine + theophylline and to leucine + theophylline by a significant increase in insulin secretion. By contrast, in cells cultured on PSSO3Na microbeads, the increase in this secretion was only slight or nil. All these abnormalities were reversible. Thus, when cells cultured on microbeads were detached and seeded on plastic wells, normal spreading and insulin secretion were observed. Lastly, PSSO3Na beads had an acute suppressive effect on insulin secretion by cells cultured on plastic wells. This study provides an example of cell-biomaterial interaction in which cell growth is possible, but with altered cell function. PMID- 3034914 TI - Sarcoma at the site of a single hip screw. A case report. AB - We report a case of malignant fibrous histiocytoma of the hip which occurred 30 years after the insertion of a single chrome-cobalt alloy screw for a slipped femoral epiphysis. The possible aetiological association between malignant tumours and metallic implants is discussed. PMID- 3034915 TI - Femoral neuropathy as a complication of aortic surgery. AB - In a series of 1006 aortic operations for atherosclerotic occlusive or aneurysmal disease a femoral neuropathy with paresis of the quadriceps femoris muscle and sensory disturbances occurred in 34 patients or 3.4%. The femoral nerve palsy was left-sided 23 times, right-sided 9 times and bilateral twice. Twenty-nine patients had a complete recovery after 1/2 to 1 years. It is suggested that the femoral neuropathy is of ischaemic nature and caused by aortic clamping. The scant blood supply of the intrapelvic section of the femoral nerve is derived from the iliolumbar artery, a branch of the internal iliac artery, and from the deep circumflex iliac artery, a branch of the external iliac artery. On the right side the deep circumflex iliac artery gives more branches to the nerve and there are more anastomoses with the fourth and fifth lumbar arteries, than on the left side. This may be the explanation for the preference of the nerve palsy for the left side. PMID- 3034916 TI - Alpha-actinin-containing aggregates in transformed cells are highly dynamic structures. AB - Normal rat kidney cells infected with a Rous sarcoma virus (strain LA23) were used to study the dynamics of alpha-actinin-containing aggregates in transformed cells. Experiments were performed by microinjecting living cells with iodoacetamidotetramethylrhodamine alpha-actinin and allowing the fluorescent analogue to incorporate into cellular structures. Subsequent time-lapse recording indicated that the alpha-actinin-containing aggregates can undergo rapid formation, movement, and breakdown. In addition, experiments using the photobleaching recovery technique indicated that alpha-actinin molecules associated with the aggregates have a very high rate of exchange, whereas those associated with adhesion plaques in normal cells exchange much more slowly. The dynamic properties of alpha-actinin-containing aggregates may be closely related to the changes in cellular behavior upon oncogenic transformation. PMID- 3034917 TI - Changes of nerve growth factor synthesis in nonneuronal cells in response to sciatic nerve transection. AB - The intact sciatic nerve contains levels of nerve growth factor (NGF) that are comparable to those of densely innervated peripheral target tissues of NGF responsive (sympathetic and sensory) neurons. There, the high NGF levels are reflected by correspondingly high mRNANGF levels. In the intact sciatic nerve, mRNANGF levels were very low, thus indicating that the contribution of locally synthesized NGF by nonneuronal cells is small. However, after transection an increase of up to 15-fold in mRNANGF was measured in 4-mm segments collected both proximally and distally to the transection site. Distally to the transection site, augmented mRNANGF levels occurred in all three 4-mm segments from 6 h to 2 wk after transection, the longest time period investigated. The augmented local NGF synthesis after transection was accompanied by a reexpression of NGF receptors by Schwann cells (NGF receptors normally disappear shortly after birth). Proximal to the transection site, the augmented NGF synthesis was restricted to the very end of the nerve stump that acts as a "substitute target organ" for the regenerating NGF-responsive nerve fibers. While the mRNANGF levels in the nerve stump correspond to those of a densely innervated peripheral organ, the volume is too small to fully replace the lacking supply from the periphery. This is reflected by the fact that in the more proximal part of the transected sciatic nerve, where mRNANGF remained unchanged, the NGF levels reached only 40% of control values. In situ hybridization experiments demonstrated that after transection all nonneuronal cells express mRNANGF and not only those ensheathing the nerve fibers of NGF-responsive neurons. PMID- 3034918 TI - Ligand- and weak base-induced redistribution of asialoglycoprotein receptors in hepatoma cells. AB - The receptor for asialoglycoproteins (ASGPR) was localized in human hepatoma Hep G2 cells by means of quantitative immunoelectron microscopy. Without ligand added to the culture medium, we found 34% of the total cellular receptors on the plasma membrane, 37% in compartment of uncoupling receptor and ligand (CURL), and 21% in a trans-Golgi reticulum (TGR) that was defined by the presence of albumin after immuno-double labeling. A small percent of the ASGPR was associated with coated pits, the Golgi stacks, and lysosomes. After incubation of the cells with saturating concentrations of the ligand asialo-orosomucoid (ASOR), the number of cell surface receptors decreased to 20% of total cellular receptors, whereas the receptor content of CURL increased by a corresponding amount to 50%. The ASGPR content of TGR remained constant. In contrast, after treatment of the cells with 300 microM of the weak base primaquine (PMQ), cell surface ASGPR had decreased dramatically to only 4% of total cellular receptors whereas label in the TGR had increased to 42%. ASGPR labeling of CURL increased only to 47%. The labeling of other organelles remained unchanged. This affect of PMQ was independent of the presence of additional ASOR. Implications for the intracellular pathway of the ASGPR are discussed. PMID- 3034919 TI - Membranes of sorting organelles display lateral heterogeneity in receptor distribution. AB - This study describes the distribution of an intrinsic membrane protein, the asialoglycoprotein receptor (ASGP-R) in the trans-Golgi reticulum and compartment of uncoupling receptor and ligand (CURL) of rat liver cells. Using quantitative immunogold electron microscopy and membrane length measurements, we showed lateral nonhomogeneity of receptors in the membranes of trans-Golgi reticulum and CURL, in particular in the membranes of secretory vesicles (identified by their content of albumin and very low density lipoprotein particles) and of CURL vesicles (endosomes), including multivesicular bodies. The characteristic tubulovesicular morphology of both sorting organelles defines the transition of receptor-rich tubular membrane and the receptor-poor limiting membrane of the attached vesicles. There was a direct relationship between the size of the secretory and CURL vesicles and the density of ASGP-Rs in their membranes. Receptor density in the smallest vesicles was similar to that found in adjacent continuous tubules. The larger the vesicles, the less receptor was detectable in their membranes. We propose that the receptor molecules are excluded from the vesicle membranes by dynamic lateral redistribution. Nonrandom receptor distribution in the CURL vesicle membranes was present even at the multivesicular body stage. These observations strongly suggest the existence of barriers to ASGP R diffusion at the junctions of tubules and vesicles. In addition, our observations suggest that ASGP-Rs are transported to the plasma membrane via a mechanism other than the normal secretory pathway. PMID- 3034920 TI - Chemically and virally transformed cells able to grow without anchorage in serum free medium: evidence for an autocrine growth factor. AB - BA10-IR transformed cells, obtained by treating Syrian hamster embryo fibroblasts (HEF) with 7-methylbenz(a)anthracene and cultivated for a long period, are highly tumorigenic and grow in suspension as aggregates (spheroids) (Levy et al., 1976). They also grow in attached form or as spheroids in serum-free (S-) synthetic medium, without insulin and transferrin, and form anchorage-independent (AI) colonies in this same, but semi-solid, medium. This exceptional phenotype was acquired stepwise, after other transformation parameters, and appears to be related to the capacity of the transformed cells to respond to a mitogenic growth factor which they secrete. The response to this autocrine factor is amplified by insulin and transferrin. Untransformed HEF, at late and early passages, and also mouse and rat embryo fibroblasts, secrete factors equally active on BA10-IR cells; but HEF do not respond, in S- medium, to their factor, or that of BA10-IR cells. Rat FR3T3 fibroblasts transformed by Kirsten murine sarcoma virus (FR3T3 Ki cells) also form AI colonies in semi-solid S- medium, secrete an autocrine factor potentiated by insulin and transferrin, and respond to the factors active on BA10-IR cells. However, they form far fewer colonies without additives, and respond as well to the mitogenic factors only in the presence of insulin and transferrin. BA10-IR cells and FR3T3-Ki cells also release beta-TGF, or a related factor, in an active and a latent form, activable by acidification, and HEF latent, activable beta-TGF. However, the factors shed by BA10-IR cells or HEF which stimulate AI growth of BA10-IR and FR3T3-Ki cells are proteins which seem unrelated to known transforming growth factors. Two major cellular alterations characteristic of the transformed phenotype in vitro are the ability to grow in the absence of anchorage, in semi-solid medium, and reduced dependence on serum growth factors (Hanafusa, 1977; Tooze, 1980). These alterations are often expressed together, and anchorage independence also appears to be the in vitro transformation parameter which correlates best with the tumorigenicity of the transformed cells (Pollack et al., 1975; Shin et al., 1975; Cifone and Fidler, 1980). However, this correlation is not constant (cf., Tooze, 1980). The cellular changes which confer anchorage independence remain unknown, but the culture conditions which allow anchorage-independent (AI) growth are better known. This growth occurs in the same media which permit the growth of attached cells, but generally requires serum.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3034921 TI - Functional differentiation of virgin mouse mammary epithelium in explant culture is dependent upon extracellular proline. AB - Depletion of proline from insulin, hydrocortisone, and prolactin-containing medium prior to incubating virgin mouse mammary explants prevents both DNA synthesis and functional differentiation in the mammary epithelial cells; however, DNA synthesis in the mammary stroma and total incorporation of radioactive amino acids into total protein appears to continue without hindrance. Removal of glycine instead of proline had no deleterious effect on either DNA replication in the hormone-stimulated epithelium or in its functional differentiation. Functional differentiation was determined by the induction of casein and alpha-lactalbumin synthesis in the insulin, hydrocortisone, and prolactin (IFPrl)-treated explant cultures. As a control, the induction of mouse mammary tumor virus (MMTV) gene expression, a corticosteroid-regulated function, was also measured. Neither the absence of proline or glycine prevented the glucocorticoid stimulation of MMTV gene expression. In contrast to mammary tissue from virgin mice, explants from nonpregnant primiparous mice responded fully to IFPrl stimulation with respect to DNA, casein, and alpha-lactalbumin synthesis in medium depleted of proline. These data suggest that the uncommitted epithelium of virgin mouse mammary glands requires the presence of exogenous proline in order to respond to lactogenic hormonal signals. We have demonstrated earlier that DNA synthesis is a prerequisite of functional differentiation in virgin mouse mammary explants (Smith and Vonderhaar, 1981, Dev. Biol., 88:167-179; Vonderhaar and Smith, 1982, J. Cell Sci, 53:97-114), although cytological differentiation proceeded unencumbered in explants prevented from synthesizing DNA. Here, without proline, neither cytological nor functional differentiation can be induced; this suggests that proline provides an essential metabolic interlock in the acquisition of lactogenic hormone responsiveness in uncommitted mouse mammary tissue. PMID- 3034922 TI - The role of benzodiazepine receptors in the induction of differentiation of HL-60 leukemia cells by benzodiazepines and purines. AB - A series of benzodiazepines was evaluated for their capacity to induce the differentiation of HL-60 acute promyelocytic leukemia cells. Benzodiazepines were effective initiators of maturation in the concentration range of 50 to 150 microM. The possible involvement of benzodiazepine receptors in mediating the differentiation induced by these agents was investigated. The presence of high affinity, peripheral type benzodiazepine binding sites (KD = 7.3 nM, TB = 14.5 pmol/mg protein with Ro5-4864) was demonstrated in HL-60 membranes. The occupancy of peripheral type high affinity benzodiazepine receptors by various benzodiazepines showed some correlation (r = 0.76) with their differentiation inducing capabilities, but binding potencies were 1,000-fold higher than the concentrations required to produce differentiation. A class of benzodiazepine receptors with lower binding affinity was also detected in HL-60 membranes (KD = 28.6 microM; TB = 199 pmol/mg protein with diazepam). A higher level of correlation (r = 0.88) was demonstrated between benzodiazepine occupancy of these lower affinity receptors and the capacity to induce maturation. Significantly, benzodiazepine concentrations needed for low affinity binding and induction of differentiation were the same (25-200 microM), suggesting that low affinity benzodiazepine receptors may be involved in the induction process. We have shown that the molecular form responsible for the induction of the differentiation of HL-60 cells to mature forms by 6-thioguanine (TGua) is the free base, TGua, itself [Ishiguro, Schwartz, and Sartorelli (1984) J. Cell. Physiol., 121:383 390]. Since hypoxanthine (Hyp) and inosine (Ino) have been identified as putative endogenous ligands for high affinity benzodiazepine receptors in brain tissue, the potential involvement of benzodiazepine receptors in the differentiation of HL-60 cells by the purines was investigated. Physiological purines such as Hyp and Ino were inactive in displacing the benzodiazepines from their high and low affinity binding sites in HL-60 membranes. In contrast, TGua caused inhibition of benzodiazepine binding to high and low affinity sites. The inhibition of Ro5-4864 binding to high affinity binding sites by TGua appeared to be due to the binding of TGua to membranes through the formation of a mixed disulfide between the 6 thiopurine and protein thiols, since the inhibition was reversed by the presence of 2-mercaptoethanol. The findings suggest a possible relationship between the occupancy of benzodiazepine receptors by TGua and the induction of leukemic cell differentiation. PMID- 3034923 TI - Thrombin chemotactic stimulation of HL-60 cells: studies on thrombin responsiveness as a function of differentiation. AB - Thrombin, a major procoagulant enzyme and growth factor, is also selectively chemotactic for monocytes and macrophages but not for neutrophils. This effect stands in contrast to other well-known chemotactic agents such as fMet-Leu-Phe, C5a fragments, and LTB4, which stimulate directed cell movement in both cell types, and have important physiological implications. The human leukemic cell line HL-60, which is capable of differentiating either along granulocytic or monocytic lineages, was therefore used to explore the development of this selective monocyte/macrophage chemotactic response to thrombin. Esterolytically inactive DIP-alpha-thrombin, as well as the thrombin-derived chemotactic peptide CB67-129, elicits a dose-dependent chemotactic response in HL-60 cells differentiated to monocytelike cells by treatment with 1,25(OH)2D3 (HL-60/mono), whereas no such response is evident in either undifferentiated HL-60 cells or in cells differentiated into granulocytes by treatment with DMSO (HL-60/gran). Similarly, early events which characterize stimulation of inflammatory cells by chemotactic agents are also evident, but only in monocyte-differentiated cells. In HL-60/mono, thrombin selectively stimulates rapid cytosolic Ca2+ elevation as well as rapid cytoskeletal association of cytosolic actin. Following thrombin stimulation, maximal actin association in these cells occurs within 30 sec (declining to basal levels at the end of 5 min), and maximal Ca2+ elevations are also evident within 15-20 sec, suggesting a temporal relationship between these two events. Thus, the events accompanying stimulation of HL-60/mono by thrombin are characteristic of those seen following stimulation of inflammatory cells by chemotaxins, with a major difference being the selectivity of thrombin as a chemotaxin for cells of macrophage/monocytic lineage. The selective chemotactic responsiveness of HL-60/mono to thrombin appears to relate to the development of specific receptors on these cells as part of monocytic differentiation: HL 60/mono (but HL-60/gran nor undifferentiated HL-60) are capable of significant specific 125-I-labeled alpha-thrombin-binding (ka approximately 20 nM), and possess an estimated 400,000 thrombin-binding sites per cell. Our findings further suggest that the thrombin response of HL-60 and particularly the expression of thrombin receptors on these cells may serve as a useful model system for exploring the biology of monocyte/macrophage differentiation. PMID- 3034925 TI - Tumors involving the skin of the upper extremity. AB - This review can only introduce the subject of tumors found involving the skin of the upper extremity. Many benign masses as well as some malignant tumors have to be considered when a patient calls to the physician's attention a lump, firm area, color change, ulcer, or other alteration in the skin. In response, the physician must have a high index of suspicion, take a careful history, and carry out a thorough examination in order to develop a safe approach. Thought has to be given to the complex anatomy of this area. Understanding of the pathophysiology of tumors and of possible later additional therapy is needed to plan an appropriate biopsy. In the brief discussions of treatment, the difficulty in choosing margins of resection and assessing the efficacy of lymph node dissection is mentioned. An open mind and assessment of future reports of studies in progress may be helpful. Whatever treatment is applied to the malignant tumors under consideration, it is my opinion that one must persist in this until one obtains tumor-free margins. The surgeon undertaking this responsibility must apply the same tumor techniques including operating room discipline that would be applied to any serious malignancy. Consultation and careful work with colleagues that are able to assess the potential for response to chemotherapy, immunotherapy, and/or radiotherapy, should be sought. Subtle hazards in our environment, such as changing risk of sun exposure, industrial chemicals, and irradiation should be pointed out to our patients. These are a challenge to the student of this subject, just as tobacco products are to those involved with malignancies of the head and neck, respiratory, and other systems. All of the previously mentioned methods must be used in the anatomically complex upper extremity to preserve function while ridding the patient of the burden of a disfiguring or painful benign process, or even a life-threatening malignancy. PMID- 3034924 TI - Characterization of the receptor for protease nexin-I:protease complexes on human fibroblasts. AB - Fibroblasts as well as several other cell types, secrete a number of protease inhibitors into their culture media. Among these inhibitors are the protease nexins, a class of proteins which covalently bind serine proteases, thereby inactivating their specific targets. Protease nexin-I, first discovered in human foreskin fibroblasts, binds thrombin, plasmin, and urokinase with high affinity, forming covalently linked complexes. Human fibroblasts bind complexes of protease nexin-I and its target protease via a cell-surface, high-affinity receptor. We have analyzed a number of characteristics of this receptor, and found them to be typical of class II receptors in general. At 4 degrees C binding of PN-I:protease complexes was competed by heparin. In addition, binding was independent of the particular protease bound to the PN-I; purified complexes of PN-I with thrombin or urokinase competed equipotently for [125]I-thrombin:PN-I binding. As the pH of the binding buffer was lowered, binding to cells increased. A twofold increase in binding was attained by lowering the pH from 7.5 to 4.5. This phenomenon was not due to irreversible, pH-induced changes to either the cell surface or the labeled complexes. At 37 degrees C, the removal of labeled complexes from culture medium was rapid; approximately 80% was removed by 4 hours under given conditions. The internalization of complexes was also very rapid, with an estimated ke (endocytic rate constant) of 1.0 min-1. At neutral pH, fibroblasts bind complexes in a saturable manner. Scatchard analysis yields a receptor number of 250,000 per cell and a Kd of 1 nM. PMID- 3034926 TI - Vascular tumors of the hand and forearm. AB - Although blood vessel tumors are rare, they are frequently encountered in the hand and forearm, being the fourth most common tumor of the hand. The treating physician should be aware of the acquired, traumatic, and congenital vascular tumors that are so prevalent in this area. PMID- 3034927 TI - Synovial sarcoma of the hand. AB - Synovial sarcoma of the hand is a very rate tumor in an equally unusual location with a difficult histology of a fibrosarcoma combined with synovial-like areas that resemble joint endothelial lining. It has a poor prognosis prior to the current combined treatment of surgical ablation, radiation to the surgical field, and chemotherapy for microscopic metastases. It has an unexplained natural history of very late recurrence and distant spread, often 10 to 20 years following primary treatment. PMID- 3034928 TI - Soft tissue sarcomas of the hand. AB - The most common soft tissue sarcomas of the hand are epithelioid sarcoma, rhabdomyosarcoma, synovial sarcoma, fibrosarcoma, and clear cell sarcoma. The epidemiology and biology of these tumors are discussed and important unifying concepts, such as local recurrences and regional lymph node metastases, are stressed. PMID- 3034929 TI - Malignant fibrous histiocytoma of the hand. AB - Malignant fibrous histiocytoma of the hand is a unique tumor in a rare location with a characteristic histology of fibrous material in a storiform pattern that is diagnostic in appearance. It has a second type of tumor consisting of histiocytic appearance without fibrous tissue. It is most difficult to treat and often requires ablative surgery combined with postoperative radiotherapy and chemotherapy for long-term survival. PMID- 3034930 TI - Primary bone tumors of the hand and carpus. AB - This discussion has attempted to outline a fundamental approach to the radiographic analysis of solitary lesions of the hand and wrist skeleton. The initial evaluation invariably relies on the routine radiograph, using the conventional film-screen combination to obtain a sense of the lesion in its gross anatomic setting. Decisions to use ancillary procedures and the selection of their type and sequence can also best be based on an initial general evaluation. On the basis of an overall view so obtained, and by using a body of general roentgenographic criteria, one can arrive at the most likely diagnostic probabilities. However, in addition to careful study of the lesion, close cooperation between the clinician, the pathologist, and the radiologist cannot be overemphasized. Only by means of such collaboration is it possible to reach a truly valid diagnosis and a rational plan for treatment. PMID- 3034931 TI - Tyrosyl and phosphatidylinositol kinases of human erythrocyte membranes. AB - The tyrosyl kinase and phosphatidylinositol (PI) kinase activities of human red cells have been partially purified and characterized. Although the PI kinase required detergent for solubilization, the major tyrosyl kinase of the red cell could be extracted by high salt. A very small residual activity remained associated with the membranes, however, that was solubilized with the PI kinase and copurified through an ammonium sulfate precipitation and diethylaminoethyl (DEAE) ion-exchange step gradient elution. However, the two activities were found to differ with respect to their apparent KmS for ATP and Mg2+; they showed different half-lives for temperature inactivation, possessed different relative activities in the presence of Mn2+ and Ca2+, and were separable by elution from a DEAE-Trisacryl ion exchange column using a linear NaCl gradient. The kinetic parameters of the membrane-associated tyrosyl kinase differed from those of the salt-extracted enzyme. PI kinase was not activated by pretreatment with the tyrosyl kinase p68v-ros or by addition of the phosphotyrosyl phosphatase inhibitor, vanadate, to intact membranes, and was not competitively inhibited by the tyrosyl kinase substrate poly(Glu4, Tyr). We conclude that the human red cell phosphatidylinositol and tyrosyl kinases are distinct and separate activities, and that at least two separable tyrosyl kinases are present in human erythrocytes. PMID- 3034934 TI - Oncogene amplification and chromosomal abnormalities in small cell lung cancer. AB - Twelve cell lines isolated from patients with small cell lung cancer have been studied for amplification of the three characterised members of the myc proto oncogene family (c-myc, N-myc, and L-myc) and for abnormalities of chromosome 3. Ten of these lines were being studied for the first time. Ten of the 12 small cell lung cancer cell lines had amplification of one member of the myc proto oncogene family. Amplification of c-myc was observed in only one small cell lung line--a "morphological variant". One "classic" small cell lung cancer line expressed c-myc but had no obvious amplification of the gene. N-myc and L-myc were more commonly amplified than c-myc. Chromosomal abnormalities (mainly deletions) in chromosome 3 were observed in all small cell lung carcinoma cell lines examined. When the small cell lung carcinoma lines were grouped according to "classic" or "variant" characteristics, it was found that the "classics" had deletions of the short arm of chromosome 3, whereas the "biochemical variants" had deletions of the long arm of chromosome 3. The extent of the deletions varied between cell lines. For the deletion in the short arm of chromosome 3 the minimum common region of overlap was assigned to bands 3p23-3p24. PMID- 3034933 TI - Myc family of cellular oncogenes. AB - The myc family of cellular oncogenes contains three well-defined members: c-myc, N-myc and L-myc. Additional structural and functional evidence now suggests that other myc-family oncogenes exist. The overall structure and organization of the c , N-, and L-myc genes and transcripts are very similar. Each gene contains three exons: encoding a long 5' untranslated leader and a long 3' untranslated region. The proteins encoded by these myc genes share several stretches of significant homology. The conservation of sequences at the carboxyterminus of the L-myc protein suggests that it is also a DNA-binding, nuclear-associated protein. Each myc gene will cooperate with an activated Ha-ras oncogene to cause transformation of primary rat embryo fibroblasts. Characteristics of several new myc-family members are described. PMID- 3034932 TI - Cellular signal transduction and the reversal of malignancy. AB - Animal cells contain only a few defined molecular systems that transduce hormonal and growth signals from the external environment to the intracellular milieu to regulate cellular growth and differentiation. Among the most ubiquitous of these "second messenger" pathways are those utilizing cyclic AMP and phosphatidylinositide turnover. The former activates protein kinase A, while the latter leads to the activation of protein kinase C and mobilization of intracellular calcium. Lesions induced by oncogenes in signal transduction systems may be responsible for the cancerous transformation of cells. In many tumor cell lines, including some transformed by the ras and sis oncogenes, activation of protein kinase A by elevation of cyclic AMP or activation of protein kinase C by addition of phorbol esters can restore many normal aspects of growth and morphology. Such "reverse transformation" is accompanied by the phosphorylation of unique cellular proteins and alterations in the phosphoinositide cycle. Molecular mechanisms by which activation of signal transduction systems can attenuate the malignant phenotype are considered in the context of cellular growth and differentiation. PMID- 3034936 TI - [Cholangiocarcinoma associated with Caroli's disease. Apropos of a case. Review of the literature]. AB - This is a report of a rarely observed occurrence of complication in Caroli's disease, i.e. the emergence of a cholangiocarcinoma in the course of the disease. There are 20 observations reported in the literature, and the hypotheses of pathogenesis set forth are reviewed. The frequency of a malignant transformation during the course of Caroli's disease is compared to the frequencies of other hepatic-biliary congenital cystic tumours. The excision of the cyst resulted in an appreciable period of survival for the patient. PMID- 3034935 TI - Induction of granuloma-dependent angiotensin-converting enzyme and eosinophil chemotactic factor in the skin of athymic nude mice. AB - Activities of angiotensin-converting enzyme (ACE), other proteinases, and eosinophil chemotactic factor (ECF-G) are known to be elevated in hepatic hypersensitivity granulomas of thymus intact (nu/+) mice after Schistosoma mansoni infection. The enzyme activities also increase, but to a lesser degree in hepatic granulomas of athymic nude (nu/nu) mice, and ECF-G is not detectable. In this study isolated hepatic granulomas from nu/+ mice were grafted into the skin of uninfected nu/nu mice, and changes in those cellular functions were determined to examine whether the newly formed granulomas by recipient nu/nu cells acquire the functional activities as well as the histological appearance of nu/+ granulomas. ACE and ECF-G rapidly disappeared from grafted sites during the first 5 days, corresponding to loss of nu/+ cells from the graft. Reduction in activities of arylsulfatases, lysozyme, and acid phosphatase also occurred, but to a lesser extent. Recovery of ACE and ECF-G activities to the levels seen in nu/+ hepatic granulomas was observed by 14 days after grafting when nu/nu cells had accumulated in the grafts and formed new granulomas. Other enzymes increased to approximately half the levels seen in grafted donor granulomas. Circulating eosinophilia also increased. The findings indicate that nu/nu cells that accumulated in the skin grafts not only morphologically mimicked nu/+ type granulomas but also demonstrated nu/+ levels of cellular function. Analysis of skin granulomas developing in nu/+ mice after grafting of nu/+ hepatic granulomas showed the similar histology and enzymatic changes, whereas the skin sites inoculated with purified schistosome eggs alone caused neither significant histological changes nor elevation of ACE activity. PMID- 3034937 TI - The 'chronic mononucleosis' syndromes. PMID- 3034938 TI - Liquid chromatography-electrochemical detection of ferro- and ferricytochrome c at a chemically modified gold electrode. AB - Chemically modified electrodes, constructed by adsorption of 4,4' dithiodipyridine onto a polyvinylferrocene-treated gold surface, were employed for the amperometric detection of cytochrome c following size-exclusion chromatography. The electrode response was nearly reversible, permitting quantitation both of the ferro-form of the protein at +0.15 V vs. Ag/AgCl and of the ferri-form at -0.15 V. The limit of detection for the reduced species was 3 pmol injected, and the response was linear over three orders of magnitude. Using the chemically modified electrode approach, cytochrome c monitoring was sufficiently selective that the compound could be determined in human plasma pretreated only by dilution and particulate filtration. PMID- 3034939 TI - Comparative chromatography of lectins and bioactivity recovery of the immunologic hormone leukoregulin on derivatized silica and on cross-linked agarose molecular sizing high-performance liquid chromatographic matrices. AB - This investigation compares the performance of the new zirconia stabilized silica and cross-linked agarose size-exclusion matrices to Spherogel-TSK 3000 SWG silica in high-performance liquid chromatographic separation of proteins possessing a range of molecular weights present in many lymphokine preparations and in recovery of bioactivity as measured by leukoregulin proliferation inhibitory activity. Retention time versus log molecular weight of protein standards was linear from 12,500 to 290,000 on the agarose and from 32,000 to 290,000 on the other columns. Recovery of leukoregulin proliferation inhibitory activity directed against RPMI 2650 epidermoid carcinoma cells was 90% from the silica, 88% from the agarose and 35% from the zirconia stabilized silica columns. PMID- 3034940 TI - Detection of some retinoid radicals using high-performance liquid chromatography with electron spin resonance spectroscopy or electrochemical detection. AB - Radical species were detected in the incubation mixtures of some retinoids (retinoic acid, retinal, retinol and retinyl acetate) by using the spin-trapping technique. The spin-adducts were resolved by high-performance liquid chromatography on a reversed-phase column with isocratic elution and detected by electron spin resonance spectroscopy and electrochemical detection. The spin adducts were eluted in the order retinoic acid, retinol, retinyl acetate, in a similar manner to the retinoids themselves. These results suggest that the spin adducts are products of nitrosobenzene with retinoid radicals in which the retinoid radicals retain their original chemical structures. PMID- 3034941 TI - Allowance for "analyte valence" in the retention model of ion-exchange chromatography. Studies of adenosine 5'-phosphates on DEAE cellulose. AB - Quantitative expressions are derived for various retention models of the ion exchange behaviour of multivalent analytes. Their effectiveness as descriptions of experimental results is then tested by application to partition equilibrium studies of the effect of phosphate concentration on the interaction between the three adenosine 5'-phosphates and diethylaminoethylcellulose at pH 4.4. A completely general multi-state model has pointed to quantitative deficiencies in the currently accepted cooperative two-state model, which is, however, superior to a multi-state model devoid of cooperative effects. A similar situation pertains to the ion-exchange behaviour of cytochrome c on carboxymethylcellulose at pH 7.0. PMID- 3034942 TI - High-performance liquid chromatographic system for the separation of tricyclic antidepressant and related drugs using ODS-Hypersil. PMID- 3034943 TI - cDNA probes of individual genes of human rotavirus distinguish viral subgroups and serotypes. AB - The use of cDNA probes for detection of rotaviruses has been investigated using plasmids containing inserts specific for each of the eleven genes of human rotavirus strain Wa. In a dot-blot detection system in which radioactive DNA probes were hybridized to viral RNA extracted from cultivatable rotavirus strains, cDNAs of genes 7, 8, 10 and 11, were found to be the most reliable probes for detecting a range of rotavirus strains. Unexpectedly, rotaviruses could be distinguished with respect to subgroup and subtype specificities when cDNAs of genes 6 and 9, which encode the immunologically relevant proteins VP6 (group-specific antigen) and VP7 (type-specific antigen), were used as probe, even though the nucleic acid sequences of these genes are known to have a high degree of sequence homology. PMID- 3034945 TI - Demonstration of hepatitis A virus in cell culture by electron microscopy with immunoperoxidase staining. AB - Virus-like particles were demonstrated by electron microscopy in BS-C-1 cells infected with hepatitis A virus (HAV). Particles were usually enclosed within vesicles and accompanied by myelin-like membranous structures. Less often they were seen free in the cytoplasm. They were never observed in the nucleus. By immunoperoxidase staining particles were found to contain HAV antigens. These antigens were also found in the membrane of the vesicles surrounding the masses of particles and adjacent parts of the mitochondrial membranes. Our results demonstrate the usefulness of an electron microscopic immunocytochemical technique to study replication of HAV. PMID- 3034944 TI - Preparation of nasopharyngeal secretions for immunofluorescence by one-step centrifugation through Percoll. AB - A simple method was developed for separation of cells from nasopharyngeal secretion for the diagnosis of respiratory virus infections by immunofluorescence microscopy. The diluted specimen, containing dithiothreitol to break up the mucus, was centrifuged once through a cushion of 20% Percoll (colloidal silica, a density gradient medium), which permitted sedimentation of cells through the cushion, but retained mucus on top of it. The pelleted cells were resuspended, and microscope slides were then prepared by standard techniques. The Percoll centrifugation method was also applicable for sputum and bronchoaveolar lavation specimens. Immunofluorescent antibody staining of nasopharyngeal secretions prepared by the described method was more sensitive than enzyme immunoassay for the detection of respiratory syncytial virus. PMID- 3034946 TI - Rapid diagnosis of herpes simplex virus infections in conventional and shell vial cell cultures using monoclonal antibodies. AB - One hundred specimens for herpes simplex virus (HSV) isolation were tested in parallel by conventional and by centrifugation-enhanced cell culture, followed by identification using monoclonal antibodies to HSV-1 and HSV-2. Sensitivity was comparable by the two methods; conventional culture was only marginally slower and was easier to fit into the routine of a busy laboratory. It is, therefore, advocated for HSV detection in clinical specimens. PMID- 3034947 TI - Aldosterone receptors in different types of primary hyperaldosteronism. AB - The number of mineralocorticoid-binding sites on mononuclear leukocytes and plasma aldosterone (aldo) concentrations were measured in patients with different types of primary hyperaldosteronism. Patients with unilateral adenoma and patients with bilateral adrenal hyperplasia had a significantly lower (P less than 0.001) mean number of binding sites for aldo [144 +/- 36 (+/- SD; n = 6) and 140 +/- 28 sites/cell (n = 4), respectively] compared with normal subjects (292 +/- 110 sites/cell; n = 25). In four patients with dexamethasone-suppressible hyperaldosteronism, mineralocorticoid-binding sites in mononuclear leukocytes were normal (291 +/- 108 sites/cell). In all patients undergoing surgery for unilateral adenoma, the receptors normalized 3 months after the operation. In two patients the reduction in receptors persisted for a short time after surgery even though the plasma aldo level had already normalized. We conclude that mineralocorticoid excess produces down-regulation of mineralocorticoid receptors, which, in turn, might contribute to the genesis of the aldo escape phenomenon. PMID- 3034948 TI - Skin-derived fibroblasts respond to human parathyroid hormone-like adenylate cyclase-stimulating proteins. AB - Human tumors and keratinocyte-conditioned medium contain PTH-like adenylate cyclase-stimulating proteins. Human dermal fibroblasts have receptors that recognize PTH and a factor associated with humoral hypercalcemia of malignancy in rats. We examined 10 human dermal fibroblast lines for an adenylate cyclase response to PTH. Six of 10 lines tested displayed a definite response (2.4- to 3.8-fold over basal) to 10(-6) M bovine PTH-(1-34). This response was inhibited by the PTH analog and antagonist Nle8,18,Tyr34-bPTH-(3-34). We also examined whether human dermal fibroblasts are capable of responding to either a human PTH like tumor-derived factor or the PTH-like factor contained in human keratinocyte conditioned medium. Both human humoral hypercalcemia of malignancy-associated tumor extract (2.5 X 10(-10) M) and keratinocyte-conditioned medium (8 X 10(-10) M) stimulated human dermal fibroblast adenylate cyclase. These concentrations are markedly lower than those required for PTH-induced adenylate cyclase stimulation. This activity was also inhibited by the PTH analog. The high prevalence of PTH responsive adenylate cyclase in dermal fibroblast lines and the apparent potency of tumor-derived and keratinocyte-derived PTH-like factors in dermal fibroblasts suggest that these factors may play a role in normal dermal physiology. PMID- 3034949 TI - In vitro study of cultured human insulinoma cells: evidence of abnormal sensitivity to glucose. AB - Human insulinoma cells were isolated and cultured in vitro, and their functional and morphological characteristics were determined. The cells, isolated as single cells or small cell clusters, reaggregated to almost the size of islets by the fifth culture day and were maintained in vitro for more than 1 month. Morphologically (light and electron microscopies) they were intact throughout the culture period. Immunohistochemically more than 50% of the cells in each reaggregate contained insulin. Incubation experiments revealed that a low glucose concentration (15 mg/dL) was sufficient to produce maximal insulin release. In the absence of glucose, 1 microgram/mL glibenclamide increased insulin release. On the other hand, 5 mM theophylline and 10 mM arginine did not alter insulin release significantly. Theophylline, arginine, and glibenclamide did not have any stimulatory effect on insulin release in the presence of 50 mg/dL glucose. Perifusion experiments with 50 mg/dL glucose disclosed a biphasic pattern of insulin release, and no significant change in insulin release occurred when the glucose concentration in the perifusate was switched from 50 to 150 and then back to 50 mg/dL. These findings demonstrate that human insulinoma cells can be isolated and maintained in vitro and that the cells have abnormal sensitivity to glucose. PMID- 3034950 TI - Influence of infused hypertonic saline on the response to insulin-induced hypoglycemia in man. AB - We studied the influence of a hypertonic saline infusion on the counterregulatory response to insulin-induced hypoglycemia in nine normal men. When given hypertonic saline, the men had less hypoglycemia in response to insulin, both acutely and in the recovery phase (P less than 0.01), and released 34% more glucagon (P less than 0.05) than when they were water loaded. The total integrated ACTH, cortisol, epinephrine, norepinephrine, and GH responses to hypoglycemia were similar after saline and water loading. After the saline load, the mean plasma vasopressin level rose from 11.0 +/- 2.2 (+/- SEM) to 20.9 +/- 2.9 pg/mL in response to insulin-induced hypoglycemia. In contrast, after the water load, vasopressin levels were undetectable (less than 2 pg/mL) and they increased only to 2.6 +/- 0.4 pg/mL with hypoglycemia. There was a significant positive correlation between basal plasma vasopressin and nadir glucose concentrations and a significant negative correlation between basal plasma vasopressin and the integrated fall in glucose after insulin administration (P less than 0.01 and P less than 0.025, respectively). The difference in the glycemic response to insulin may be related to the high vasopressin levels after saline loading, which could, either directly and/or through enhanced glucagon release, increase hepatic glucose production and thus limit the hypoglycemic response to insulin. PMID- 3034951 TI - The roles of Ca2+ and adenosine 3',5'-monophosphate in the regulation of progesterone production by human placental tissue. AB - The effects of Ca2+ and cAMP on progesterone production by placental tissue were studied. Term placentas obtained from normal pregnant women were perfused with sterile saline to remove blood, and the minced trophoblastic tissue was incubated in vitro. The rate of progesterone secretion by the trophoblastic tissue was 47.5 +/- 5.0 (+/- SE) ng/mg cell protein X h (control) at 450 micrograms low density lipoprotein/mL. The rates of progesterone production and cAMP accumulation were accelerated 3.0- and 1.7-fold, respectively, by 10(-5) M terbutaline, and the terbutaline effect was blocked by the addition of 50 microM N-(6-aminohexyl)5 chloro-1-naphthalenesulfonamide (W-7) or 50 microM trifluoperazine. Whereas progesterone secretion by placental explants cultured in medium containing low density lipoprotein with 1 microM A23187 (calcium ionophore) reached a rate of 128.5 +/- 8.5 ng/mg cell protein X h, incubation of placental explants with A23187 caused a highly significant, dose-related decrease in terbutaline stimulated cAMP accumulation in the presence of 3-isobutyl-1-methylxanthine, a phosphodiesterase inhibitor. Inhibition by A23187 was Ca2+ dependent, since incubation in a Ca2+-free medium with EGTA blocked its effect. Intracellular Ca2+ is apparently necessary for placental progesterone production; enhancement of progesterone production by beta2-stimulation is mediated by intracellular Ca2+ and cAMP. PMID- 3034952 TI - 24-hour profiles of adrenocorticotropin, cortisol, and growth hormone in major depressive illness: effect of antidepressant treatment. AB - Plasma ACTH, cortisol, and GH concentrations were measured at 15-min intervals for 24 h in 11 men suffering from major depressive illness during an acute episode of depression and during clinical remission following antidepressant treatment with either electroconvulsive therapy or amitriptyline. Seven age matched normal men also were studied. During the acute phase of the illness, the patients had abnormally short rapid eye movement sleep latencies, hypercortisolism, early timing of the nadirs of the ACTH-cortisol rhythms, and shorter nocturnal periods of quiescent cortisol secretion. GH was hypersecreted during wakefulness, and a major pulse occurred before, rather than after, sleep onset. After treatment, rapid eye movement sleep latencies were lengthened, and cortisol levels returned to normal due to a decrease in the magnitude of episodic pulses. Moreover, the timing of the circadian rhythms of ACTH and cortisol as well as the duration of the quiescent period of cortisol secretion were normalized. The amount of GH secreted during wakefulness decreased to normal values, with fewer significant GH pulses. The major elevation of GH secretion in the early part of the night occurred later than that during the depressive episode. These results demonstrate that a disorder of circadian rhythmicity characterizes acute episodes of major depressive illness and that this chronobiological abnormality as well as the hypersecretion of ACTH, cortisol, and GH are state rather than trait dependent. PMID- 3034953 TI - Effects of suramin on the function and structure of the adrenal cortex in the cynomolgus monkey. AB - There have been reports of adrenal failure in patients treated with suramin, an agent that has recently been used as therapy for acquired immune deficiency syndrome. We conducted this study to assess the effect of suramin on adrenal function and structure in a primate, the cynomolgus monkey. Five male monkeys were treated with suramin (800 mg/m2, im) once a week, for 5 weeks. Five other animals (controls) received saline. The treated animals had progressive elevations of plasma ACTH (P less than 0.05) and PRA (P less than 0.02) and decreased serum cortisol responses 30 min after the administration of synthetic ACTH (P less than 0.05) compared to controls. There was disruption of the architecture of the adrenal cortex, a diffuse inflammatory cell infiltrate, and thinning of the zona glomerulosa and zona fasciculata in the suramin-treated animals. We conclude that suramin is toxic to adrenal cortical tissue and might be useful in treating conditions with adrenal cortical hyperfunction, such as adrenal cortical carcinoma and Cushing's syndrome. PMID- 3034954 TI - Hormonal regulation of messenger ribonucleic acids for P450scc (cholesterol side chain cleavage enzyme) and P450c17 (17 alpha-hydroxylase/17,20-lyase) in cultured human fetal adrenal cells. AB - ACTH has acute and long term effects on adrenal steroidogenesis by week 14 of fetal life. We used human fetal adrenal cells to investigate the long term effect of physiological doses of ACTH on mRNAs for P450scc (the cholesterol side-chain cleavage enzyme) and P450c17 (17 alpha-hydroxylase/17,20-lyase). Monolayer cultures of 18- to 24-week gestation fetal zone adrenal cells were maintained in the presence and absence of 10(-9) or 10(-8) M ACTH for up to 12 days. As assessed by RNA dot blots probed with cloned homologous human cDNAs, ACTH increased P450scc and P450c17 mRNAs 4- and 9-fold, respectively, over control values on day 7 of culture. ACTH-mediated stimulation was slightly less on day 12 of culture. The ACTH-mediated accumulation of those mRNAs were time dependent. When cells were exposed to a single 10(-8)-M dose of ACTH, the amount of P450scc and P450c17 mRNA was increased by 24 h, reaching a maximum at 48 h and diminishing by 72 h. When cells were maintained in 10(-8) M ACTH continuously, mRNA for both enzymes accumulated in a similar pattern, reaching a peak at 48 h but remaining at nearly maximal values thereafter, up to 96 h. Dibutyryl cAMP (10(-3) M) mimicked these stimulatory actions of ACTH, although its effect was greater at 24 h and more stable up to 96 h. Angiotensin II (1-100 ng/mL) and hCG (1-100 ng/mL) had no effect on accumulation of P450scc and P450c17 mRNAs. The production of both dehydroepiandrosterone sulfate and cortisol also was stimulated by ACTH, suggesting that the increased mRNAs were translated into active enzymes. These results indicate that ACTH induces human fetal adrenal cells to accumulate mRNAs for both P450scc and P450c17; this effect of ACTH is probably mediated by cAMP. Chronic 96-h stimulation of human fetal adrenal cells did not diminish their responsiveness to ACTH. Together with our earlier studies of the human fetal adrenal, these data indicate that fetal adrenal tissue does not exhibit the desensitization to trophic hormone stimulation characteristic of adult tissue. PMID- 3034955 TI - Effect of upright posture on the aldosterone responses to dopamine, metoclopramide, angiotensin II, and adrenocorticotropin. AB - Aldosterone secretion in man is stimulated by potassium (K), ACTH, and angiotensin II (AII) and inhibited by dopamine (DA). In normal sodium-replete supine individuals, aldosterone secretion is under maximum tonic inhibition by DA and is not inhibited further by DA administration. Sodium depletion alters plasma aldosterone responses to secretogogues. Upright posture, another physiological stimulus to aldosterone secretion, recently was demonstrated to sensitize the adrenal cortex to inhibition of aldosterone secretion by a large quantity of DA (4.0 micrograms/kg X min). The effect of upright posture on aldosterone responses to other secretogogues is unknown. In this study, we investigated the effect of upright posture on aldosterone responses to low infusion rates of DA, to the DA antagonist metoclopramide (M) and to AII and ACTH. Fourteen normal men eating a normal sodium diet were studied. In eight, PRA, plasma aldosterone (PAC), plasma cortisol (F), and serum K concentrations were determined after 4 h of upright posture and infusion of vehicle (D5W) or DA at 0.1, 0.4, and 2.0 micrograms/kg X min. Six other normal men were kept supine for 3 h and, on separate days, upright for 3 h and given iv M (10-mg bolus dose), AII (1 and 4 pmol/kg X min for 30 min), and ACTH (20 and 120 mU/h for 30 min). PAC, PRA, F, and K were measured before and after these three secretogogues were administered. In the presence of vehicle, mean PAC increased by 15.1 +/- 4.3 (+/- SEM) ng/dL after 4 h of upright posture. In the presence of DA infused at 0.1, 0.4, and 2.0 micrograms/kg X min, the PAC response to upright posture was decreased to 9.7 +/- 2.5 (P = NS), 7.5 +/ 3.9 (P less than 0.05), and 8.1 +/- 2.0 (P less than 0.05) ng/dL, respectively. This occurred without a decrease in PRA, F, or K. The stimulation of PAC 10 and 20 min after a 10-mg bolus dose of M was 9.6 +/- 3.3 and 9.3 +/- 2.6 ng/dL, respectively, in supine subjects and 8.3 +/- 2.3 and 10.8 +/- 3.4 ng/dL 10 and 20 min after the M dose in upright subjects. The responses of PAC to ACTH and AII also were unchanged after 3 h of upright posture. We conclude that upright posture sensitizes the adrenal cortex to inhibition of aldosterone secretion by DA without affecting other modifiers of aldosterone secretion.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3034956 TI - Dissociation between cortisol and adrenal androgen secretion in patients receiving alternate day prednisone therapy. AB - To evaluate the hypothesis that chronic, low dose, alternate day prednisone treatment may suppress adrenal androgen secretion without causing long term suppression of the hypothalamic-pituitary-adrenal axis we studied seven patients with systemic lupus erythematosus who had been taking low dose (5-20 mg), alternate day prednisone therapy for at least 1 yr. Basal and ovine CRH (oCRH) stimulated plasma ACTH, cortisol, and adrenal androgen levels were measured 12 h (day on) and 36 h (day off) after the most recent dose of prednisone, and the results were compared to those in seven age- and sex-matched normal subjects. The patients' basal ACTH and cortisol levels did not differ significantly from those in the normal subjects on either the day on or the day off prednisone treatment. By contrast, their basal adrenal androgen levels were significantly decreased compared to those in normal subjects on both the day on and the day off prednisone (P less than 0.05). The patients' oCRH-stimulated ACTH and cortisol levels on the day off prednisone did not differ from normal levels, but were significantly blunted during the day on prednisone (P less than 0.05). In contrast, the patient's oCRH-stimulated adrenal androgen levels were significantly decreased during both the day off and the day on prednisone (P less than 0.05). These findings are consistent with the hypothesis that chronic alternate day prednisone therapy, at doses close to or below replacement, suppresses adrenal androgen levels without long term suppression of the hypothalamic-pituitary-adrenal axis. Based upon these findings, we postulate that an alternate day regimen of prednisone might maintain the benefits while reducing the risks of glucocorticoid therapy of adrenal hyperandrogenism. PMID- 3034958 TI - Visualization of somatostatin receptors and correlation with immunoreactive growth hormone and prolactin in human pituitary adenomas: evidence for different tumor subclasses. AB - Using combined autoradiographic and immunohistochemical techniques, specific somatostatin (SRIH) receptors were measured in 31 human pituitary tumors. SRIH receptor distribution was compared with that of immunoreactive GH and PRL in the same tissue sections. Among the 10 tumors from acromegalic patients, 7 contained a high or medium density of SRIH receptors, whereas only a low density of such receptors was present in the remaining 3 tumors despite a comparably high concentration of immunoreactive GH in all 10 tumors. No correlation between plasma GH levels and SRIH receptors was found. One corticotroph tumor was SRIH receptor negative. Among the 5 prolactinomas tested, only 1 contained a significant density of SRIH receptors, the distribution of which did not correlate with that of immunoreactive PRL cells. Interestingly, among the 15 endocrine-inactive adenomas investigated, 6 had a measurable, sometimes high, density of SRIH receptors despite undetectable GH, PRL, or TSH immunoreactivity. These results suggest that based on their SRIH receptor content, there may be subclasses of GH-producing pituitary adenomas. The difference in SRIH receptor density may be an explanation for the clinically described differential responsiveness of acromegalic patients to SRIH. The results also suggest that subclasses of endocrine-inactive pituitary adenomas may exist; some of them contain significant amounts of SRIH receptors, which may be responsive to SRIH treatment. PMID- 3034957 TI - Pituitary response to intravenous hypothalamic releasing peptides in cynomolgus monkeys treated with contraceptive steroids. AB - The secretory response of the pituitary to an iv bolus dose of hypothalamic releasing peptides (HRP) was evaluated in male and cycling (CYC) or contraceptive treated female cynomolgus monkeys. Parenteral delivery of levonorgestrel and 17 beta-estradiol by intravaginal ring (CVR) was compared with oral administration (OC) of norgestrel and ethinyl estradiol in the diet. LH secretion was suppressed in the CVR group compared to that in other groups, and the response in males was greater than that in either CYC or OC females (P less than 0.01). Elevated plasma PRL concentrations in the CVR group during the baseline period (P less than 0.05) together with their larger pituitary weights (P less than 0.01) suggested lactotroph hypertrophy or hyperplasia compared to other groups. The plasma GH response was similar in the male, CYC, and CVR groups, but plasma GH levels increased from -15 to 0 min before HRP injection in the OC group (P less than 0.001) and continued to be higher for 15 min after HRP compared to values in the other groups (P less than 0.001), suggesting a treatment effect. Neither plasma TSH nor T4 levels were different among the groups after HRP administration, but T4 was elevated (P less than 0.01) in the OC group due to increased T4-binding globulin. The greater ACTH response 15 min after HRP treatment (P less than 0.05) in the CVR group compared to that in the other groups was associated with greater adrenal weights of the CVR females (P less than 0.05), suggesting chronic tropic stimulation. However the adrenal steroid results did not support this interpretation. We conclude that the differences in pituitary hormone secretion during these studies could be attributed to the nature of the reproductive steroid environment. PMID- 3034959 TI - Activation, arousal, and neuropsychological rehabilitation. AB - This paper addresses the concept of arousal and the role of the reticular system in producing the orienting response. For patients with deviant autonomic responses, the use of central neuropharmacological stimulants is considered. Their therapeutic value is discussed in relation to hyperactive children and individual adult cases of the posttraumatic syndrome consequent upon traffic accidents. PMID- 3034960 TI - Pathology of trophoblastic diseases. AB - This paper furnishes a synoptic overview of the anatomic pathology of trophoblastic growths, with some reference to clinical implications. I will attempt to provide definitions and generalizations; however, the limitation of this approach is all too familiar. Definitions are constructed to cover the borderline case, but the borderline, as if by definition, escapes. For every generalization there are exceptions, and any pathologist who has assembled a large series of cases of a given disease entity is acutely aware of the exceptions he has encountered. Likewise, classifications are an artifact we superimpose on nature in an attempt to make order from experience and to rationalize it. It can be awkward, if not unfair, to shove the square peg of the biologic case into the round hole of man-made taxonomy. PMID- 3034961 TI - Hydatidiform mole after in-vitro fertilization: a case report. AB - A 25 year old patient was accepted on the in-vitro fertilization programme at Groote Schuur Hospital in Cape Town and her first IVF cycle resulted in a pregnancy. The patient did not return to the usual follow-up ultrasound until about 4 months after the embryo transfer when an ultrasound revealed the typical picture of a molar pregnancy which was confirmed by histology after evacuation. PMID- 3034962 TI - Detection of cytomegalovirus infections in specimens other than urine by the shell vial assay and conventional tube cell cultures. AB - Blood, bronchoscopy-lavage, biopsy (lung, liver, kidney), sputum, and other (cecum, bone) specimens were inoculated into shell vials and conventional cell tube cultures seeded with MRC-5 cells over a 23-month period. Of 1,472 specimens, 182 (12.4%) yielded cytomegalovirus (CMV)-positive results from 81 patients. Significantly more CMV-positive specimens were detected in shell vials (n = 154; 84.6%) than in conventional tube cell cultures (n = 126; 69.2%) (P less than 0.01). We found that 98 (53.8%) of the total 182 and 41 (42.7%) of the 96 blood specimens positive for CMV were detected by both the shell vial assay and conventional tube cell cultures. However, 56 (30.7%) of the total 182 and 31 (32.3%) of the 96 blood specimens positive for CMV were obtained exclusively in shell vials after detection with monoclonal antibody. Alternatively, 28 (15.4%) of the total 182 and 24 (25%) of the 96 blood specimens positive for the virus were isolated only in conventional tube cell cultures. Thus, although the shell vial assay was more sensitive and rapid than the conventional tube cell culture method, both systems must be used, especially for blood specimens, for the laboratory diagnosis of CMV infections. PMID- 3034963 TI - Comparison of genomic, plasmid, synthetic, and combined DNA probes for detecting Plasmodium falciparum DNA. AB - Total genomic Plasmodium falciparum DNA, the plasmid clone pRepHind, and a 21 base-long synthetic DNA probe (PFR1), the sequence of which was derived from pRepHind, were hybridized with DNA from various species of the phylum Apicomplexa. The genomic probe hybridized with P. reichenowi and P. falciparum DNA and significantly cross-hybridized with DNA of all the other Plasmodium species tested. The synthetic and plasmid probes hybridized to P. falciparum DNA and at reduced levels to P. reichenowi but did not hybridize to P. vivax, P. malariae, P. ovale, P. fragile, P. inui, P. knowlesi, Babesia bovis, B. microti, B. bigemina, Anopheles sp., Pan sp., Aotus sp., or human DNA. Southern blot analysis indicated that approximately 60 distinct restriction enzyme fragments from P. falciparum DNA were similarly detected by PFR1 and pRepHind. A method was developed by using a second brief hybridization with synthetic DNA to amplify signals from samples that were previously hybridized with plasmid-borne repetitive DNA. This amplification procedure was shown to allow the detection of 0.005% P. falciparum parasitemias from 10-microliter samples of blood from patients in Kenya. PMID- 3034965 TI - Detection of feline leukemia virus infection in saliva. AB - The question was investigated whether feline leukemia virus (FeLV) infection may be diagnosed by testing saliva in an enzyme-linked immunosorbent assay (ELISA). Saliva was collected with commercially available swabs, eluted from the swabs, and tested in the ELISA. A comparison of results with saliva and serum samples from 60 specific-pathogen-free cats, 9 experimentally infected cats, and 1,117 field cats led to the following conclusions. False-positive saliva results, if any occurred, were rare events. During experimental infections, antigen excretion in saliva was observed 1.5 weeks after the first appearance of FeLV antigen in serum. In one of four positive serum samples from sick animals brought to veterinarians, saliva samples tested negative. The use of saliva in an ELISA for the detection of FeLV p27 in individual sick cats is therefore less reliable than the use of serum. In seven cats with diseases typical of FeLV, including one with an intestinal form of lyphosarcoma, saliva tested positive and serum tested negative. Based on the saliva and serum results for cats living in 92 multicat households, it was concluded that saliva may be a useful secretion for FeLV screening. PMID- 3034964 TI - Detection of antibody to group B adult diarrhea rotaviruses in humans. AB - Group B rotaviruses have been responsible for annual epidemics of severe diarrhea affecting both adults and children in China. We developed a specific and sensitive enzyme-linked immunosorbent blocking assay to detect antibody to group B rotaviruses that will be useful to assess the role of group B rotavirus infections as a cause of human gastroenteritis. We tested 219 human sera and 18 immunoglobulin pools collected from eight countries for antibodies to both group A and group B rotaviruses. Overall, a low proportion (10 of 237 or 4.2%) of sera contained antibody to group B rotaviruses. Antibody to group B rotavirus was detected in only 1 of 155 serum samples from healthy or hospitalized individuals in the United States, including patients with the chronic inflammatory bowel diseases Crohn's disease and ulcerative colitis. No antibody was detected in 15 serum samples from Australia and from an outbreak of gastroenteritis on a cruise ship or in nine immunoglobulin pools from Japan and the United Kingdom. Antibody to group B rotaviruses was detected in 8 convalescent-(but not acute-)phase serum samples from Chinese patients with group B gastroenteritis, in five immunoglobulin pools from China, in 1 of 6 serum samples from Chinese students in the United States, and in 1 each of 10 serum samples from Kenya, 20 from Thailand, and 15 from Canada. In contrast, most of these samples (226 of 237 or 95.4%) had antibody to group A rotaviruses. These results indicate that human infection with group B rotavirus has not been widespread in areas outside China. Seroconversion observed between the acute-and convalescent-phase serum samples from China also suggests that infections with this virus are primary infections. Continued surveillance for this new group of rotaviruses should determine whether the many susceptible people become infected of whether other factors influence the severe pathogenicity of human infections with these viruses in China. PMID- 3034966 TI - Comparison of fluorescent-antibody-to-membrane-antigen test, indirect immunofluorescence assay, and a commercial enzyme-linked immunosorbent assay for determination of antibody to varicella-zoster virus. AB - The demand for sensitive and specific assays to determine immune status to varicella can be expected to increase with the anticipated availability of a varicella-zoster virus vaccine for use in nonimmune adults, especially health care personnel, and in immunosuppressed children. Although the fluorescent antibody-to-membrane-antigen (FAMA) test remains the reference standard to which other tests are compared, simpler alternative assays are needed. In this study, the FAMA was compared with a simple indirect immunofluorescence assay (IFA) and a commercially available enzyme-linked immunosorbent assay (ELISA) for the detection of antibody to varicella-zoster virus. One hundred and twelve serum samples were screened by the FAMA test and IFA at a 1:5 dilution, and 100% agreement was found. Of these samples, 101 were available for testing by ELISA, and identical results were obtained with 97 samples (96% agreement). When the samples were screened at a 1:2 dilution, 99 of 101 results agreed. In addition, 31 spinal fluid samples were tested by all three methods. When screening was at a 1:2 dilution, there was 96.8% agreement between the FAMA test and IFA. When the cutoff value established for sera was used for the spinal fluid samples, there was 90.3% agreement between the ELISA and the FAMA test. Thus, both IFA and ELISA can be considered sensitive and specific alternatives to the FAMA test, and in addition, both use commercially available reagents. PMID- 3034967 TI - Evaluation of a rapid latex slide agglutination test for herpes simplex virus as a specimen screen and culture identification method. AB - A total of 449 clinical specimens and 199 culture fluids were tested using the Virogen Herpes Slide Test (Wampole Laboratories, Div. Carter-Wallace, Inc., Cranbury, N.J.), a rapid latex agglutination procedure. The results were compared with those obtained with isolation of herpes simplex virus in cell culture followed by identification using immunoperoxidase or fluorescent reagents. The sensitivity, specificity, and positive and negative predictive values of the direct test were 49.7, 93.4, 96.0, and 37.1%, respectively. The sensitivity and specificity of the latex agglutination test for culture confirmation were 75.9 and 100%, respectively. PMID- 3034968 TI - Simultaneous seeding and infecting of shell vials for rapid detection of cytomegalovirus infection. AB - Fifty cytomegalovirus isolates were used to infect shell vials containing confluent MRC-5 cell monolayers and shell vials which were seeded with MRC-5 cells at the time of inoculation. All 50 of the isolates were detected by immunofluorescence in shell vials containing confluent monolayers, whereas 39 (78%) of the isolates were detected in shell vials that had been seeded and inoculated simultaneously (P less than 0.001). Preformed monolayers of cells in shell vials provide the most sensitive system for the detection of cytomegalovirus infection. PMID- 3034969 TI - Isolation of group A swine rotaviruses displaying atypical electropherotypes. AB - Swine rotaviruses displaying distinctive electropherotypes were isolated from the feces of diarrheic piglets in two swine herds in the province of Buenos Aires, Argentina. In one case all samples isolated showed abnormal electropherotypes. All samples were classified as group A reactive when assayed by an enzyme-linked immunosorbent assay. Three samples from this herd were adapted to grow in tissue culture. The electrophoretic pattern of the genomic RNA as well as the group A reactivity of one of these viruses was retained after cloning in MA-104 cells. In the other pig unit were found samples displaying both classical and abnormal electropherotypes. These viruses were also positive in the enzyme-linked immunosorbent assay; however, since they could not be adapted to grow in tissue culture, this classification must be considered tentative. The abnormal electropherotype exhibited by these pig viruses strongly resembles those of human origin called super short. PMID- 3034970 TI - Detection of herpes simplex virus in direct specimens by immunofluorescence assay using a monoclonal antibody. AB - A monoclonal antibody (MAb), designated CHA 437, was developed against herpes simplex virus (HSV). This MAb (isotype, immunoglobulin G2b K) reacted with HSV type 1 and HSV type 2. It showed no cross-reactivity with varicella-zoster virus, cytomegalovirus, or Epstein-Barr virus. Direct detection of HSV antigen in clinical specimens using indirect immunofluorescence with this MAb was compared with tissue culture isolation. For the 682 specimens tested, the direct specimen test gave a sensitivity of 84.6% and a specificity of 95.7%. PMID- 3034971 TI - Prevalent patterns of serotype-specific seroconversion in Mexican children infected with rotavirus. AB - The level of neutralizing antibodies to rotaviruses belonging to serotypes 1, 3, and 4 was determined in acute- and convalescent-phase sera from 36 Mexican children with rotaviral diarrhea. Most of the infants who seroconverted fell into one of the following three patterns: single seroconversion to serotype 1; seroconversion to serotypes 1 and 4; or seroconversion to all three serotypes tested. The heterotypic neutralizing antibody responses to rotavirus infections are discussed. PMID- 3034972 TI - Silica gel as transport medium for Corynebacterium diphtheriae under tropical conditions (Indonesia ) AB - Silica gel was confirmed as a useful transport medium for Corynebacterium diphtheriae in the investigation of diphtheria cases in which there is no ready access to laboratory facilities. PMID- 3034973 TI - (Na+ + K+)-ATPase in C6 glioma and rat cerebrum. AB - The content and distribution of the membrane-bound enzyme (Na+ + K+)-ATPase in a rat cerebral C6 glioma was determined by immunocytochemistry, immunoblots and enzyme assay. In the C6 glioma cell culture (Na+ + K+)-ATPase activity was about 20% of (Mg2+ + Na+ + K+)-ATPase activity. However, (Mg2+ + Na+ + K+)-ATPase activity in the cerebral C6 gliomas was very close to Mg2+ baseline and not significantly increased by Na+ and K+. As shown by immunoblotting, (Na+ + K+) ATPase catalytic subunit was detected in excised samples of control cerebrum and as a trace in the intracerebral portions of C6 glioma but not at all in the extracranial portions of C6 glioma or in C6 glioma cell culture. (Na+ + K+) ATPase was not detected immunocytochemically in paraffin sections of the extracranial or intracerebral portions of rat cerebral C6 glioma. The absence of staining for (Na+ + K+)-ATPase clearly demarcated projections of glioma within normal brain. These results suggest that C6 glioma has little if any expression of (Na+ + K+)-ATPase in vitro or in vivo. The small amount of enzyme epitope in the intracerebral portions represents contamination by normal cerebrum in the extracts. PMID- 3034974 TI - Protection of C58 mice from lactate dehydrogenase-elevating virus-induced motor neuron disease by non-neutralizing antiviral antibodies without interference with virus replication. AB - The paralytic poliomyelitis induced in old, immunosuppressed C58 mice by a primary infection with the lactate dehydrogenase-elevating virus (LDV) is prevented by the presence of anti-LDV antibodies in the virus inoculum or by passive transfer of LDV-free plasma from chronically LDV-infected mice one day before infection. Non-neutralizing antibodies were protective and specifically directed to the lowest molecular weight form of the envelope glycoprotein of LDV (VP-3), which seems to exist in virions in at least ten molecular forms ranging from 24 to 44 kDa. The antibodies did not prevent the productive infection of the subpopulation of macrophages that represents the primary permissive cell type in the mouse as evidenced by normal plasma LDV levels nor the spread of LDV to the central nervous system. Many non-neuronal cells containing LDV RNA were detected by in situ hybridization in the spinal cords of mice that had been infected with LDV in the presence of protective antibodies. However, no LDV RNA-positive neurons were detected, which are normally found coincidental with the development of paralytic symptoms in LDV-infected C58 mice. We propose that an early event after infection is critical for the infection of neurons and is inhibited by the presence of non-neutralizing antibodies to the LDV glycoprotein. PMID- 3034975 TI - Role of somatosensory evoked potentials in the diagnosis of peripheral nerve lesions: recent advances. AB - Short-latency somatosensory evoked potentials (SEPs) have become an important tool in the investigation of peripheral nerve lesions. Electrically evoked SEPs are most suitable because they provide results with small variations and are readily repeatable. Techniques for testing 10 different upper and lower limb nerves, dermatomes, cutaneous fibers of trigeminal nerve, and nerves supplying urogenital areas are now available. The established, principal areas of application are in the investigation of brachial plexus lesions, proximal injuries of individual nerves in upper and lower limbs, and painful dysesthesias and in the differential diagnosis of pain caused by psychogenic causes or organic lesions. The techniques have proven to be of value in demonstrating early proximal abnormalities in polyneuropathies. Abnormalities have occurred in trigeminal nerve lesions, urogenital dysfunctions, hereditary ataxias, and rare neuropathies. Unexpected abnormalities have also been reported in motor neuron disease, myotonic muscular dystrophies, and other conditions. Applications in the diagnosis of spondylopathic root lesions are not satisfactory; the techniques' usefulness in the investigation of these lesions does not extend beyond aiding the selection of patients for other diagnostic modalities, such as myelography, computed tomography (CT), and nuclear magnetic resonance (NMR) imaging. The usefulness of SEP techniques in the diagnosis of peripheral nerve lesions will remain, even if advances in organ imaging techniques can provide more specific information about the level and magnitude of lesions in the central nervous system. SEP techniques are an important complement to the other well-established methods, such as clinical testing, electromyography, and nerve conduction studies. PMID- 3034976 TI - Hematopoietic growth factors. PMID- 3034977 TI - Sodium-potassium pump, ion fluxes, and cellular dehydration in sickle cell anemia. AB - We studied the role of the sodium-potassium pump in erythrocytes of 12 patients with sickle cell anemia (SS). Ouabain-binding sites per cell and pump-mediated Rb/K uptake were significantly higher in SS patients than in white or black controls. Ouabain-resistant Rb/K influx was also greater than in normal controls or patients with sickle cell trait. Deoxygenation of SS erythrocytes increased ouabain-sensitive Rb/K influx without altering ouabain binding, presumably as the consequence of an increase in the passive influx of sodium. Deoxygenation increased mean corpuscular hemoglobin concentration (MCHC) by 5.5%, and studies of the density distribution of SS cells indicated an increase in highly dense fractions known to contain sickled erythrocytes. Ouabain prevented the rise in MCHC and reduced the percentage of dense cells. These findings indicate a magnified role for the sodium-potassium pump in the pathophysiology of SS erythrocytes and suggest that its inhibition might prove useful in therapy. PMID- 3034978 TI - myc family oncogene amplification in tumor cell lines established from small cell lung cancer patients and its relationship to clinical status and course. AB - 44 small cell lung cancer cell lines established from 227 patients were studied for myc family DNA amplification (c-myc, N-myc, and L-myc). Two of 19 lines (11%) established from untreated patients' tumors had DNA amplification (one N-myc and one L-myc), compared with 11 of 25 (5 c-myc, 3 N-myc, and 3 L-myc) cell lines (44%) established from relapsed patients' tumors (P = 0.04). The 19 patients who had tumor cell lines established before chemotherapy treatment survived a median of 14 wk compared with 48 wk for the 123 extensive stage patients who did not have cell lines established (P less than 0.001). Relapsed patients whose cell lines had c-myc DNA amplification survived a shorter period (median of 33 wk) than patients whose cell lines did not have c-myc amplification (median of 53 wk; P = 0.04). We conclude that myc family DNA amplification is more common in tumor cell lines established from treated than untreated patients' tumors, and c-myc amplification in treated patients' tumor cell lines is associated with shortened survival. PMID- 3034980 TI - Granulocyte-macrophage colony-stimulating factor. Sensitive and receptor-mediated regulation by 1,25-dihydroxyvitamin D3 in normal human peripheral blood lymphocytes. AB - We show that 1,25-dihydroxyvitamin D3 (1,25[OH]2D3), the most hormonally active metabolite of vitamin D3, modulates sensitively and specifically both the protein and messenger RNA accumulation of the multilineage growth factor granulocyte macrophage colony-stimulating factor (GM-CSF). The regulation of GM-CSF expression is seen in both normal human mitogen-activated T lymphocytes and T lymphocytes from a line (S-LB1) transformed with human T cell lymphotropic virus 1 (HTLV-1). In contrast, cells from a HTLV-1 transformed T lymphocyte line (Ab VDR) established from a patient with vitamin D-resistant rickets type II with undetectable 1,25(OH)2D3 cellular receptors are resistant to the action of 1,25(OH)2D3. Inhibition of GM-CSF expression by 1,25(OH)2D3 can occur independently of interleukin 2 regulation and is probably mediated through cellular 1,25(OH)2D3 receptors. We conclude that 1,25(OH)2D3 may be important in the physiology of hematopoiesis. PMID- 3034981 TI - Vitamin D3 and cardiovascular function in rats. AB - We have previously identified a receptor for 1,25-dihydroxyvitamin D3 in myocardial cells (Simpson, R.U. 1983. Circulation. 68:239.). To establish the relevance of this observation, we evaluated the role of the prohormone vitamin D3 in regulating cardiovascular function. In rats maintained on a vitamin D3 deficient diet for nine weeks, increases in systolic blood pressure (BP) and serum creatine phosphokinase (CPK) were observed. These increases coincided with a reduction of serum calcium from 10.3 to 5.6 mg/dl. However, while serum calcium remained depressed throughout the study, increases in BP and serum CPK were transient. After nine weeks of vitamin D3-depletion, but not after six weeks, ventricular and vascular muscle contractile function were also markedly enhanced. The increase in ventricular contractile function could not be prevented by maintaining serum calcium at 9.0 mg/dl during the period of D3-depletion. These observations suggest a primary role for the vitamin D3-endocrine system in regulating cardiovascular function. PMID- 3034979 TI - Oxidant-mediated epithelial cell injury in idiopathic pulmonary fibrosis. AB - Lung inflammatory cells of patients with idiopathic pulmonary fibrosis (IPF) were evaluated for their ability to injure 51Cr-labeled AKD alveolar epithelial cells in the presence and absence of IPF alveolar epithelial lining fluid (ELF). The IPF cells were spontaneously releasing exaggerated amounts of superoxide (O.2) and hydrogen peroxide (H2O2) compared with normal (P less than 0.02). Cytotoxicity of the AKD cells was markedly increased when the IPF inflammatory cells were incubated with autologous ELF (P less than 0.02). The majority of IPF patients had ELF myeloperoxidase levels above normal (P less than 0.002). Incubation of IPF ELF with AKD cells in the presence of H2O2 caused increased cellular injury (P less than 0.01 compared with control), which was suppressed by methionine, a myeloperoxidase system scavenger. IPF patients with high concentrations of ELF myeloperoxidase deteriorated more rapidly than those with low ELF myeloperoxidase (P less than 0.05). Thus, IPF is characterized by an increased spontaneous production of oxidants by lung inflammatory cells, the presence of high concentrations of myeloperoxidase in the ELF of the lower respiratory tract, and a synergistic cytotoxic effect of alveolar inflammatory cells and ELF on lung epithelial cells, suggesting oxidants may play a role in causing the epithelial cell injury of this disorder. PMID- 3034984 TI - Inhibitor of the factor VIIa-tissue factor complex is reduced in patients with disseminated intravascular coagulation but not in patients with severe hepatocellular disease. AB - Inhibition of Factor VIIa-tissue factor activity by a plasma component(s) that requires factor Xa has been described recently. In this communication, we have developed a specific radiometric assay (which utilizes 3H-Factor IX and is sensitive to less than 1% of plasma level) for this inhibitor and have measured its activity in various disease states. Strikingly, the levels of this inhibitor were found to be normal in patients with advanced chronic hepatocellular disease but low in patients with disseminated intravascular coagulation (DIC). When endotoxin was used to induce DIC in rabbits, the levels of this inhibitor fell by 25-90%. Human umbilical vein endothelial cells (HUVE), bovine pulmonary artery endothelial cells, and a human hepatoma cell line (HepG2) all synthesized and secreted this inhibitor, whereas a promyelocytic cell line (HL-60) did not and a monocytic cell line (U937) appears to synthesize only small amounts. When ammonium sulfate-fractionated human plasma and serum-free conditioned media from both HUVE and HepG2 cells were electrophoresed on sodium dodecyl sulfate acrylamide gels, two activity peaks corresponding to Mr approximately 45,000 and Mr approximately 33,000 were eluted in each case. These observations suggest that (a) the inhibitor is consumed in DIC and that (b) endothelial cells (or other cells) synthesize sufficient amounts of this inhibitor in vivo to compensate for any decreased production by liver cells. PMID- 3034985 TI - Peripheral thrombocytopenia in human parvovirus infection. PMID- 3034982 TI - Glomerulonephritis induced in the rabbit by antiendothelial antibodies. AB - The effects of interaction between endothelial angiotensin converting enzyme (ACE) and goat anti-rabbit ACE (GtARbACE) antibodies were studied in rabbit glomeruli. By immunofluorescence ACE was not detectable in normal glomeruli. However, when kidneys were perfused with GtARbACE antibodies glomerular bound IgG was seven times higher than that of non-immune IgG and granular deposits of goat IgG were found on the endothelium of glomeruli and arteries. Rabbits injected intravenous for 4 d with GtARbACE antibodies showed on day 1 granular deposits of goat IgG on the glomerular endothelium; from day 3 to 24 there was gradual development of subepithelial deposits of goat IgG, rabbit IgG and C3. When GtARbACE antibodies were similarly injected into proteinuric rabbits there was formation of subepithelial granular deposits of goat IgG and ACE. The results document that a glomerular endothelial antigen is redistributed in vivo by a specific ligand, an event associated with formation of immune deposits. Furthermore, if the glomerular permeability is artificially increased, immune complexes shed from nonglomerular endothelia into the circulation can contribute to form subepithelial immune deposits. PMID- 3034983 TI - Changes in catecholamine-induced lipolysis in isolated human fat cells during the first year of life. AB - Catecholamine-induced lipolysis in isolated human adipocytes during the first year of life was investigated. During this period fat cell size increased markedly. Basal and catecholamine-induced glycerol release were positively correlated with age when lipolysis was expressed per cell. However, when lipolysis was expressed per unit of cell surface area (micrometer squared), this correlation was observed only for noradrenaline. Basal lipolysis and the effect of the pure beta-agonist, isoprenaline, were identical in infants and adults. From 0 to 2 mo of age noradrenaline had very little lipolytic effect. The addition of the alpha-2-adrenoceptor antagonist, yohimbine, to noradrenaline equalized lipolysis per micrometer squared in infants and adults and the alpha-2 adrenoceptor sensitivity was significantly enhanced in infants. In both groups the lipolytic adrenoceptor was of the beta-1 type. In conclusion, adipocytes from infants have a poor lipolytic response to noradrenaline partly because of the small fat cells but mainly because of an enhanced alpha-2-adrenoceptor activity. PMID- 3034986 TI - Cytomegalovirus (CMV) antibody screening in blood donors: modification of new latex agglutination test compared with two standard methods. PMID- 3034987 TI - Intra-axonal labeling of saccular afferents in the goldfish, Carassius auratus: correlations between morphological and physiological characteristics. AB - Lucifer yellow was used to label axons of 32 physiologically defined S1 afferent fibers and 23 physiologically defined S2 afferent fibers of the goldfish sacculus. Analysis of these labeled fibers allowed us to study the relationship between electrical activity in these primary neurons and the morphology of their peripheral arborizations in the sensory macula. Morphological characteristics were highly indicative of response type: The peripheral arborizations of individual S1 fibers occupied a relatively small region (approximately 40 microns in diameter) in the rostral one-fourth of the saccular macula, whereas those of individual S2 fibers covered a larger area (roughly 80 microns across) in the caudal part of the macula. In addition to this rostrocaudal dimension, a ventral projection was related to a rarefaction response, a dorsal projection was related to a compression response, and a two-sided innervation was related to fibers having both responses. S1 fibers had either large or small terminals; the S2 fibers had only small terminals. On average, S1 fibers gave rise to approximately four terminals (e.g., seven small terminals or one to two large terminals) and S2 fibers approximately ten terminals. Spontaneous discharges were absent in all S1 fibers but present in some S2 fibers. Such S2 fibers showed spontaneous activity of either an irregular type or a burst type. In these, there was a tendency for fibers having more extensive arborizations to exhibit a burst type of spontaneous discharge. We conclude that structure-function relationships can be determined for these primary neurons. PMID- 3034988 TI - Demarcations of the mechanosensory projection zones in the raccoon thalamus, shown by cytochrome oxidase, acetylcholinesterase, and Nissl stains. AB - To determine anatomically the boundaries and internal organization of the kinesthetic and cutaneous mechanosensory regions of the ventrobasal thalamus, alternate section series from electrophysiologically mapped tissues from 14 raccoons were stained for cytochrome oxidase, myelinated fibers, acetylcholinesterase, and Nissl substance. Microelectrode tracks, along with electrolytic lesions placed as tissue markers, reveal that the mechanoreceptor projection zones have higher cytochrome oxidase and lower acetylcholinesterase staining than some neighboring regions. Both these enzymatic stains reveal particularly sharp boundaries separating the mechanoresponsive region, from the lateral posterior nucleus dorsally and from the ventroposterior inferior nucleus ventrally. The kinesthetic projection zone is often separated from other mechanoreceptor projections by bundles as well as laminae of myelinated fibers, similar to those separating cutaneous projections from distinct body parts. These subdivisions are particularly well marked by the cytochrome oxidase stain. The combination, in neighboring sections, of the use of the several stains adds considerably to the visible delineation of these functionally distinct regions, beyond what can be seen in Nissl-stained sections. PMID- 3034989 TI - Morphology of expiratory neurons of the Botzinger complex: an HRP study in the cat. AB - In anesthetized and artificially ventilated cats, the physiological and morphological properties of expiratory neurons or their axons of the Botzinger complex (BOT) were studied using intracellular recording and intracellular HRP labeling techniques. Thirteen expiratory neurons (nine cell somata and four axons) were successfully stained. Four of them were motoneurons, having relatively large cell somata in the retrofacial nucleus (RFN) and axons without any collaterals inside the brainstem. All the motoneurons showed a plateau shape of depolarization potentials during the expiratory phase. Any of the other nine expiratory neurons exhibited augmenting type firing or membrane potential changes during the expiratory phase. In five out of nine augmenting neurons, cell somata were stained and located ventral to the RFN. In four, only axons were stained. The majority of the augmenting neurons had two major axonal branches: one traveling toward the contralateral side and the other descending ipsilaterally in the brainstem. The most striking feature of the axonal trajectory was that all of the stained augmenting expiratory neurons, including the axons, had collateral branches with synaptic boutons in the BOT area, thus indicating that BOT expiratory neurons interact with some respiratory neurons in the BOT area and its vicinity. PMID- 3034990 TI - Effect of mineral salts, carbachol, and pilocarpine on nutrient digestibility and ruminal characteristics in cattle. AB - Fifty percent concentrate diets containing 2% sodium bicarbonate, sodium chloride, or no additional mineral salts were fed at a rate of 86 g dry matter/kg body weight X 75/d to three barren Holstein cows fitted with ruminal fistulas in a 3 X 3 Latin square design. Dietary adaptation was 14 d followed by 4 d collection. Ruminal pH, liquid volume, liquid dilution rate, and particulate rate of passage were increased with dietary mineral salts. Six Holstein cows fitted with ruminal fistulas were administered .01 mg carbachol/kg body weight/d, .10 mg pilocarpine/kg body weight/d, or saline placebo via subcutaneous, osmotically controlled pumps in a replicated 3 X 3 Latin square design. Treatments were administered for a 14-d adaptation period followed by an 8-d collection period. Both carbachol and pilocarpine increased liquid dilution rate, particulate rate of passage, and percent cellulolytic bacteria, whereas liquid volume was reduced. PMID- 3034991 TI - Coxsackievirus B3 murine myocarditis: a pathologic spectrum of myocarditis in genetically defined inbred strains. AB - Group B coxsackieviruses are the most frequent causative agents in human viral myocarditis. Susceptibility to viral infections varies widely among individuals. In the mouse, coxsackievirus B3 also causes myocarditis. The differential susceptibility of different inbred strains of mice to coxsackie B3-induced myocarditis also appears to be under genetic control. This study details the histopathology of coxsackie B3 myocarditis in six different inbred strains of mice for the first 45 days after coxsackie B3 infection. These strains differ either in the haplotypes of their major histocompatibility complex or in their background genome. During the first 7 days after coxsackie B3 infection, there are dramatic differences among strains with respect to prevalence and severity of myocarditis. Focal zones of myocyte necrosis involving polymorphonuclear leukocytes as well as contraction band injury appear to be the early manifestations of direct viral injury. Four of the six strains, though, continue to show myocardial inflammation after day 9. This late phase myocarditis is characterized by the emergence of mononuclear cells within healing foci of myocyte necrosis as well as a distinctive diffuse interstitial pattern of myocarditis. The strains that develop this late ongoing myocardial inflammation frequently produce heart-specific autoantibodies. Thus the pathologic features of murine coxsackie B3 myocarditis change over the course of the illness, and genetic susceptibility to both early and late phase myocarditis differs markedly among various mouse strains. PMID- 3034993 TI - The glycemic index, fiber, and the dietary treatment of hypertriglyceridemia and diabetes. PMID- 3034992 TI - Prevention of viral myocarditis with recombinant human leukocyte interferon alpha A/D in a murine model. AB - Effects of recombinant human leukocyte interferon alpha A/D on experimental myocarditis due to encephalomyocarditis virus were investigated. Plaque reduction assays revealed that 50% of plaque formation in vitro in human amnion (FL) cells was inhibited by interferon alpha A/D (9.7 U/ml) when it was administered 24 hours before infection with the encephalomyocarditis virus. Four week old male DBA/2 mice were inoculated intraperitoneally with 10 plaque-forming units (pfu) of encephalomyocarditis virus. Interferon alpha A/D was administered subcutaneously (10(2) U/g body weight per day in Group 1, 10(3) U/g per day in Group 2 and 10(4) U/g per day in Group 3) starting 1 day before infection. It was also administered starting the same day in Group 4 and 1 day after virus inoculation in Group 5 (10(4) U/g per day in both groups). Control mice were injected with saline solution. Each group consisted of 10 mice; they were killed on day 4 for evaluation. Myocardial virus titers were significantly lower in Group 3 (8.2 +/- 25.2 X 10(2) pfu/mg, p less than 0.05) and Group 4 (3.0 +/- 5.5 X 10(3) pfu/mg, p less than 0.05) than in control mice (5.6 +/- 4.1 X 10(4) pfu/mg). Histologic examination showed extensive myocardial necrosis and cellular infiltration in all control mice, but no myocardial necrosis or cellular infiltration in Group 3 and less severe necrosis and infiltration in Group 4. There were no significant differences in myocardial virus titers or histologic changes between control mice and Group 1, 2 or 5.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3034994 TI - Evaluation of a high-fiber diet in hyperlipidemia: a review. AB - Epidemiologic studies of cardiovascular mortality rates in different countries have suggested that dietary fiber may play a protective role. Within a similar population, a large intake of fiber is associated with a lower relative risk of death from coronary heart disease. Dietary fiber may be separated into at least two types: insoluble, which includes cellulose, hemicellulose, and lignin; and soluble, including pectin and gums. Laxative effects appear to predominate with insoluble fibers such as wheat bran, with little change in plasma lipid levels in most studies. Pectin, guar gum, and oat bran (soluble fibers) have been reported to have hypocholesterolemic effects in both animals and man, with the effect being proportional to the degree of cholesterol elevation. Other gums, specifically those from locust bean and karaya, have a similar effect, with the decrease in total cholesterol due primarily to a decrease in the low-density lipoprotein cholesterol fraction. While some studies have shown continued improvement over a period of months, this has not been uniformly found. Both normal and elevated triglyceride levels appear to be more resistant to change with dietary fiber. An increase of dietary carbohydrate as a source of fiber may be associated with an increase in triglyceride levels. Fiber may, however, offer some protection against an increase in cholesterol and triglyceride levels in subjects fed diets containing large amounts of sucrose. Although rats fed oat bran, guar gum, or pectin had lower levels of hepatic and blood triglycerides, humans with hypercholesterolemia fed oat bran or guar showed no effect on their triglycerides.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3034995 TI - Evaluation and management of corticotropin allergy. AB - Subsequent use of an adrenocorticotropic hormone (ACTH) preparation may be considered in some instances in a patient who has demonstrated a suspected or proven allergic reaction to corticotropin on prior administration. We describe a patient with multiple sclerosis with a history of anaphylaxis to porcine ACTH in whom another course of treatment with ACTH was being considered. The patient had immediate cutaneous reactivity to porcine ACTH but not to the synthetic ACTH peptide, cosyntropin. With a test-dose schedule, we were able to uneventfully administer cosyntropin to the patient. The presence of serum IgE antibody directed against porcine ACTH and absence of IgE antibody against cosyntropin were demonstrated by ELISA technique, corroborating the skin test results. These studies are consistent with previous evidence that immediate hypersensitivity reactions to corticotropin are IgE mediated and support the value of skin testing in the clinical evaluation and management of known or suspected corticotropin hypersensitivity. PMID- 3034996 TI - Prooxidant action of desferrioxamine: Fenton-like production of hydroxyl radicals by reduced ferrioxamine. AB - It is common practice in biochemical research to assume that iron bound to desferrioxamine (DFO) to form ferrioxamine (FOA) has been rendered inactive to subsequent redox chemistry within the range of physiological redox potentials, both in vitro and in vivo. However, plants and microorganisms can make iron metabolically available from ferrioxamine and closely related trihydroxamate siderophores, and at neutral pH, cyclic voltammetry of FOA demonstrates a reversible one-electron reduction at about -0.42 to -0.45 V (vs. normal hydrogen electrode), which is within the range of a number of reducing enzymes. We present evidence for the Fenton-like ability of FOA reduced by paraquat cation radicals to consume H2O2 and produce hydroxyl radicals (OH.) in the process. Similar reactions may explain previously reported potentiation of the oxidizing toxicity of paraquat in rats by high doses of DFO, as well as several other examples of prooxidant actions of DFO in vivo. We present the hypothesis that biphasic antioxidant/prooxidant behavior of DFO as a function of dose may be common with iron-catalyzed oxidizing reactions when mobile strong reducing agents are present. Hence, the real possibility of amplifying oxidizing damage must be considered when planning treatment with DFO, and failure of DFO to inhibit a particular response to oxidizing stress or its enhancement by DFO cannot, by itself, be considered sufficient evidence to rule out an iron-dependent process. PMID- 3034997 TI - Calcium efflux from sarcoplasmic reticulum microsomes due to oxidation and sulfhydryl-binding agents. AB - Calcium permeability of sarcoplasmic reticulum (SR) microsomes was measured after aging or after exposure to peroxydisulfate or to sulfhydryl-binding agents. Under conditions where the Ca2+-ATPase was active, the maximum net release of Ca2+ was not significantly different between control and oxidized SR. However, when calcium uptake was prevented by EGTA or apyrase, the Ca2+ permeability of oxidized microsomes was 2 to 3 times greater than control of low (10(-9), 10(-7) M) but not high (10(-6) M) levels of external calcium. The observation that vesicles preincubated with 5 mM dithiothreitol loaded up to 3 times as much calcium and had a slightly lower calcium permeability coefficient than control vesicles suggested that sulfhydryl oxidation might modulate calcium flux. This hypothesis was tested by exposing to sulfhydryl-binding agents: silver, arsenite, and p-chloromercuriphenylsulfonic acid. Sulfhydryl-binding agents initiated a rapid release of calcium from microsomes, and release was halted by dithiothreitol. Inhibition of calcium transport could not entirely account for the apparent increase in permeability because the calcium permeability of SR treated with sulfhydryl-binding agents was 5 times greater than that of SR exposed to Ca2+-ATPase inhibitors. These results suggest that oxidation may increase the calcium permeability of SR by allowing calcium loss through a channel that can be gated by sulfhydryl oxidation. PMID- 3034998 TI - Ceruloplasmin, extracellular-superoxide dismutase, and scavenging of superoxide anion radicals. AB - Ceruloplasmin and extracellular-superoxide dismutase are similar in physical properties. Both are found in extracellular fluids and both are scavengers of the superoxide radical. The relationship between the two proteins was further explored in the present investigation. Ceruloplasmin preparations were found to be commonly contaminated with extracellular-superoxide dismutase. In one preparation, 80% of the superoxide dismutase activity was due to extracellular superoxide dismutase. Ceruloplasmin, freed from contaminating superoxide dismutase, was found to catalytically dismute the superoxide anion radical with a rate constant of about 1.0 X 10(4) M-1 s-1 per copper atom. Under physiological conditions with a low rate of superoxide production, ceruloplasmin preferentially reacts stoichiometrically with the superoxide radical with a rate constant of about 2 X 10(5) M-1 s-1 per copper atom. Under such conditions, the reaction does not result in hydrogen peroxide formation. From the kinetic data obtained it was calculated that in normal human plasma, extracellular-superoxide dismutase will scavenge about twice as much superoxide as ceruloplasmin. Using immobilized antibodies toward extracellular superoxide dismutase and ceruloplasmin, no antigenic cross-reactivity between the two proteins could be detected. PMID- 3034999 TI - Photosensitization by antitumor agents 3: spectroscopic evidence for superoxide and hydroxyl radical production by anthrapyrazole-sensitized oxidation of NADH. AB - EPR and spin-trapping techniques were employed to study the oxidation of the dihydronicotinamide adenine dinucleotide (NADH) photosensitized by an anthrapyrazole-antitumor agent. The superoxide radical was detected as a DMPO adduct upon illumination of the system with visible light. Photoinduced generation of hydroxyl radicals is demonstrated by detection of DMPO adducts of OH scavengers, such as ethyl alcohol, sodium formate, and sodium azide. The dependence of the production of these spin adducts on the presence of catalase implies the involvement of hydrogen peroxide in that process. The production of hydrogen peroxide is demonstrated independently during oxygen consumption measurements with the Clark electrode technique. PMID- 3035000 TI - Human medullary thyroid carcinoma in tissue culture; secretion of calcitonin and carcinoembryonic antigen. AB - Monolayer cultures of medullary thyroid carcinoma (MTC) cells were prepared from 5 patients with familial and 4 with sporadic MTC. Basal calcitonin (CT) release decreased rapidly during 17 days in culture. Addition of extra calcium (2 mmol/l) or dibutyryl cyclic AMP (dBcAMP, 3 mmol/l) increased CT release in all investigated cultures (p less than 0.01). No significant differences were observed between cell suspensions prepared from familial and those from sporadic MTC. Carcinoembryonic antigen (CEA) was found in 3 out of 5 MTC cell suspensions from patients with familial tumors and in 2 out of 2 sporadic tumors. Only two cultures secreted measurable amounts of CEA, which decreased with culture time but at a much slower rate than that of CT release. CEA release was not affected by the addition of calcium and dBcAMP. This study shows an absent relation between basal and stimulated CT and CEA release by cultured MTC cells. PMID- 3035001 TI - Gynecomastia associated with isolated ACTH deficiency. AB - A 57-year-old man with gynecomastia associated with isolated adrenocorticotropic hormone deficiency is reported. Basal levels of estrogens, luteinizing hormone and prolactin were elevated, whereas plasma testosterone levels were normal. Luteinizing hormone and prolactin showed exaggerated responses to gonadotropin releasing hormone and thyrotropin-releasing hormone, respectively. These hormonal abnormalities were corrected and gynecomastia resolved with steroid replacement therapy. These findings suggest that gynecomastia may result from hormonal changes which were modified by glucocorticoid deficiency. PMID- 3035003 TI - [Ante-partum administration of preventive treatment of Rh-D immunization in rhesus-negative women. Parallel evaluation of transplacental passage of fetal blood cells. Results of a multicenter study carried out in the Paris region]. AB - 1,969 non immunized rhesus negative primiparous women were followed up in 23 maternity units in the geographical region of Paris. 1,882 could be retained to study antepartum protection and 1,884 to study transplacental passage of fetal blood cells. Two groups were defined according to whether they were born in even or uneven years, so that: 955 were the "control" group who delivered 590 rhesus D positive infants, and 927 were the "treated" group who delivered 599 rhesus D positive infants. The "control" group were used as controls at the 28th and 34th weeks of pregnancy, while the "treated" group received two injections of anti-D immunoglobulin given on the same dates after taking the necessary tests. Immunological testing at the time of the delivery and after the delivery showed that 7 women had become Rh D immunized in the "control" group whereas only one in the "treated" group. This difference, which is statistically significant, confirms the results of other authors about the efficiency of antepartum rhesus disease prevention. The incidence of immunisation during or immediately after the first pregnancy in women who had no previous story of blood transfusions or of terminations of pregnancy is 1.11%, which is a figure relatively low as compared with studies of series carried out in North America, but close to those carried out in other European centres. When primipara of all categories are lumped together the frequency rises to 1.5%. A study of the passage of fetal red blood cells into the maternal circulation shows that at the 29th week of pregnancy out of 1,884 cases there were 5.5% positive kleihauer tests, without a large volume of blood being detected and at the 34th week of pregnancy when 957 tests were carried out, 7% were positive with one of them being of a massive transfusion of blood from the fetus to the mother, which was life-threatening for the fetus. It may be that the incidence had been under-estimated and that the positive results in the two groups, control and treated, show that there is a statistically significant difference that demonstrates that antepartum treatment in the trial has eliminated a worthwhile percentage of positive kleihauer tests which arose from the transfusion of small quantities of blood.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3035002 TI - Naloxone increases the response of growth hormone and prolactin to stimuli in obese humans. AB - Opiates stimulate the growth hormone and prolactin responses to stimuli in non obese humans. Obese patients, however, show lowered growth hormone and prolactin responses and raised beta-endorphin levels. We therefore investigated the effect of the opiate antagonist naloxone on the stimulated growth hormone and prolactin secretions in a controlled double-blind study in obese patients. All patients received 200 micrograms TRH and 0.5 g/kg b.w. arginine together with 2 mg of naloxone or placebo i.v. in a randomized sequence. The TRH- and arginine-induced increases in prolactin and growth hormone were significantly greater after administration of naloxone (p less than 0.05). Naloxone also produced a significant increase in ACTH, cortisol and beta-endorphin when compared with placebo. TSH, triiodothyronine, thyroxine, insulin, glucagon and blood glucose showed no significant differences between both days of the trial. The effect of naloxone on growth hormone and prolactin secretions in obese humans can thus be regarded as a partial normalization. We therefore conclude that the hypothalamic regulatory disturbance of growth hormone and prolactin secretions in the obese could be caused by raised opiate levels. PMID- 3035004 TI - Carpal tunnel release in patients with diffuse peripheral neuropathy. AB - Sixty carpal tunnel decompressions were performed in 44 patients with combined carpal tunnel syndrome and peripheral neuropathy. Symptomatic improvement was obtained in 92% of the patients and complete relief of symptoms in 72%. We conclude that peripheral neuropathy is not a contraindication to carpal tunnel decompression. PMID- 3035005 TI - Hepatitis B virus (HBV) and hepatocellular carcinoma. HBV DNA status and its implications. PMID- 3035006 TI - Electron spin resonance spectrometrical study of the melanins in the wool of some North European sheep in relation to their color inheritance. AB - Additional peaks that were known on the esr-spectrograms of red human and reddish brown Karakul hair to be diagnostic traits of phaeomelanin esr-signal also were found on esr-spectrograms of the tan, but not of black or chocolate brown wool from Icelandic sheep. This tan color is thought to depend on the presence of phaeomelanin and is due to the top dominant allele at the A locus. The two methods of distinguishing between eu- and phaeomelanin-dependent brown colors- esr-spectrometrical and genetical--are in agreement for European as well as for Asiatic breeds. Both light and dark brown Soay fleece samples lacked the additional peaks and are interpreted as eumelanin pigmentation. PMID- 3035007 TI - Fixation and immunofluorescent analysis of creatine kinase isozymes in embryonic skeletal muscle. AB - Pectoral muscles from chicken embryos of various ages were examined with immunofluorescent and radiolabeled probes for the presence of brain-type creatine kinase (B-CK), muscle-specific creatine kinase (M-CK), muscle-specific myosin heavy chain (MHC), and cycling cells. The diffusible creatine kinase isozymes were not detectable by indirect immunofluorescence after standard histological fixation of embryonic muscle. However, a fixation procedure was devised that permitted immunodetection of the creatine kinase isozymes (particularly B-CK) in embryonic tissue from all stages of development studied. B-CK, M-CK, and MHC were all detected in post-mitotic muscle cells, but only B-CK was detected in cycling cells. Correlations between these findings and in vitro observations of a deterministic muscle lineage are discussed. PMID- 3035008 TI - Angiotensin converting enzyme inhibitors: drug interactions. PMID- 3035009 TI - Antigen-specific protection against graft-vs-host induced immune deficiency. AB - A severe, antigen-nonspecific, and long-lasting immune-deficient state can be induced in healthy, adult immune-competent F1 hybrid mice by a single i.v. injection of parental T lymphocytes. The present report demonstrates that this graft-vs-host-induced immune deficiency (GVHID) can be prevented in an antigen specific way by immunization of the F1 mice with allogeneic cells before induction of GVHID. Thus, spleen cells from (A X B)F1 mice primed with allogeneic cells from strain C and then injected with parental spleen cells from A did not generate cytotoxic T lymphocyte responses to trinitophenyl-modified self cells or to allogeneic cells from third party strains D or E. However, spleen cells from the same mice generated normal levels of cytotoxic T lymphocyte activity to allogeneic cells from C, the strain used for immunization. Furthermore, mice exposed to murine cytomegalovirus before induction of GVHID were resistant to a subsequent challenge with murine cytomegalovirus, whereas GVHID mice that received only the murine cytomegalovirus challenge all died. These findings are discussed with respect to the possibilities that primed and unprimed T helper cells may be differentially susceptible to the suppressive effects of GVH. PMID- 3035010 TI - Characterization of dual-reactive H-2Kb-restricted anti-vesicular stomatitus virus and alloreactive cytotoxic T cells. AB - Cross-reactive recognition of alloantigen by "self + X"-reactive cytotoxic T lymphocytes (CTL) has been documented in a variety of systems. It has been shown previously that the H-2Kb-restricted CTL response of C57BL/6 (B6) mice to vesicular stomatitis virus (VSV) infection is partially cross-reactive on uninfected target cells expressing the H-2Kbm8 mutation. In this report, we describe the isolation and detailed characterization of such dual-reactive CTL. By employing EL4 tumor lines transfected with genes encoding various VSV proteins, we demonstrated that the majority of dual-reactive CTL recognize the internal N protein of VSV and are also reactive against uninfected bm8 targets. Although the response of normal B6 mice to bm8 stimulators shows no measurable cross-reactivity on VSV-infected targets, the response of VSV-primed B6 mice to bm8 stimulation is almost entirely cross-reactive, lysing VSV-B6 targets and uninfected bm8 targets roughly equally. Furthermore, about 70% of CTL clones isolated from such mice by bm8 stimulation are dual-reactive with respect to effector function. Analysis at the population and clonal levels with cold target competition and antibody blocking suggests that the bulk of dual-reactive CTL have a higher avidity for VSV-B6 targets than for bm8 targets. The extreme case of this is illustrated by a fraction of CTL clones, isolated and maintained on bm8 stimulators, which lyse VSV-B6 targets but do not lyse bm8 targets. One such CTL clone is shown to be specific for the bm8 antigen in proliferation assays. These results demonstrate that: the specificity of an alloreactive CTL response may be dramatically altered by previous antigenic encounters; and dual-reactive CTL display a significant difference in affinity of the CTL receptor-determinant interaction, depending on the target which is recognized. PMID- 3035011 TI - The enhancement of the immune response by pain stimulation in mice. I. The enhancement effect on PFC production via sympathetic nervous system in vivo and in vitro. AB - Effects of catecholamines and osmotical and physical stimuli on the induction of anti-sheep red blood cells (SRBC) plaque-forming cells (PFC) were investigated in (C57BL/6 X BALB/c)F1 mice in vivo and in vitro. The anti-SRBC PFC from mice immunized with 5 X 10(7) SRBC was markedly increased by daily s.c. injections of epinephrine. The enhancement of PFC by epinephrine was completely blocked by preadministration with propranolol and hexamethonium, but not with phentolamine. The PFC was increased by osmotic and physical stimuli given once a day for 4 days after immunization with SRBC. The enhancement of PFC by these stimuli was completely blocked by preadministration with propranolol and hexamethonium. The enhancement of PFC by physical stimuli was observed in nonimmunized mice when spleen cells from stimulated mice were cultured with SRBC in vitro. In normal mice, the enhancement of PFC was observed 2 hr after one physical stimulation. However, spleen cells from mice given two physical stimuli did not show the enhancement of PFC after treatment with anti-Thy-1.2 antibody and complement, nor after removal of nonadherent cells. Next, the serum obtained from mice 30 to 60 min after a physical stimulation enhanced PFC of normal mice spleen cells in vitro, but the enhancement was abolished by the addition of propranolol. The enhancement of anti-SRBC PFC by s.c. injection of epinephrine suggested that the autonomic nervous system, especially the sympathetic nervous system, was activated by a local stimulus effect of the injection. This enhancement of anti SRBC PFC appear to be due to the activation of antigen non-specific helper T lymphocytes by the beta-actin of endogenous catecholamines from the adrenal gland. PMID- 3035012 TI - Stimulation of normal inducer T cell clones with antigen presented by purified Ia molecules in planar lipid membranes: specific induction of a long-lived state of proliferative nonresponsiveness. AB - Culture of normal inducer T cell clones with antigen and purified Ek beta:Ek alpha incorporated into planar lipid membranes resulted in specific T cell activation as determined by cell volume increase and IL 3 production. However, in contrast to results obtained with T cell hybridomas, antigen presentation by planar membranes did not induce measurable IL 2 production, and proliferative responses were not detected. Rather, recognition of only Ek beta:Ek alpha and antigen resulted in the specific induction of a long-lived state of proliferative nonresponsiveness to subsequent stimulation by conventional APC and antigen. Induction of nonresponsiveness required protein synthesis, and was not simply due to the absence of IL 2. The antigen-nonresponsive cells could respond to either PMA plus ionomycin or IL 2, and they expressed normal levels of surface antigen receptor molecules. These results demonstrate that recognition by normal T cell clones of antigen and Ia molecules in the absence of other accessory cell molecules and signals results in a prolonged state of proliferative nonresponsiveness, possibly similar to a state of T cell tolerance in vivo. PMID- 3035013 TI - Immune complexes bind to cultured rat glomerular mesangial cells to stimulate superoxide release. Evidence for an Fc receptor. AB - Contractile mesangial cells (MC) possess a number of macrophage-like characteristics, including oxygen radical generation. We suggest that under certain conditions MC may serve as immune effector cells in glomerulonephritis. Immune complex (IC) deposits are a hallmark of glomerulonephritis. Because IC elicit oxygen radicals from other cell types and because oxygen radicals can induce glomerular injury, we measured release of O2- by cultured rat MC in response to IC and, in separate experiments, the binding of IC to MC. Soluble and insoluble IC markedly stimulated dose- and time-dependent, saturable O2- release. Specific antibody (Ab) alone or mixtures of nonimmune Ab and antigen had no significant effect. IC-induced O2- release was not affected by cytochalasin B, an inhibitor of phagocytosis. Binding studies with radioiodinated IC demonstrated specific binding with an affinity of 1.56 X 10(6) M-1 and 1.02 X 10(5) receptors per cell. Both binding and O2- release required the Fc region of Ab. IC formed with F(ab')2 fragments did not bind specifically to or stimulate O2- release by MC. Cultured cells from rats depleted of bone marrow-derived phagocytes by irradiation produced amounts of O2- similar to cells from normal rats. These results provide evidence that IC affect the biology of the contractile glomerular MC in a manner that is dependent on the Fc region of Ab and suggest that MC structure and function may be altered at sites of injury. PMID- 3035014 TI - Lysosomal enzyme release from human monocytes by particulate activators is mediated by beta-glucan inhibitable receptors. AB - Human peripheral blood monocytes ingest particulate activators and generate leukotrienes via a trypsin-sensitive, beta-glucan-inhibitable receptor. The incubation of monolayers of monocytes with from 4 X 10(5) to 2 X 10(8) zymosan or glucan particles resulted in a dose-dependent release of up to 9% +/- 1.9 and 17.8% +/- 5.3 (mean +/- SD, n = 3) of the lysosomal enzyme, N acetylglucosaminidase, into the culture medium. Lysosomal enzyme release occurred throughout the 2-hr period studied, with the greatest rate of N-acetyl glucosaminidase release occurring during the first hour; the presence of 5 micrograms/ml of cytochalasin B accelerated this process when zymosan was the agonist. The preincubation of monocytes with from 0.5 to 500 micrograms/ml of soluble yeast beta-glucan inhibited N-acetylglucosaminidase release by 4 X 10(7) zymosan and glucan particles in a dose-dependent manner, with 50% inhibition occurring with 50 micrograms/ml of soluble yeast beta-glucan (mean +/- SD, n = 3). Preincubation with as much as 5 mg/ml of yeast mannan had no inhibitory effect on N-acetylglucosaminidase release. The pretreatment for 30 min of monolayers of monocytes with 50 micrograms/ml of affinity-purified trypsin, which selectively inactivates the monocyte-phagocytic response to particulate activators, also fully inhibited lysosomal enzyme release induced by zymosan and glucan particles. The inhibitory effects of a soluble ligand, yeast beta-glucan, and of trypsin pretreatment on lysosomal enzyme release correspond to the inhibitory effect of these agents on monocyte phagocytosis of zymosan and glucan particles and thus indicates ligand specificity for the beta-glucan receptor in the release of stored intracellular mediators. PMID- 3035015 TI - Opioid-mediated suppression of cultured peripheral blood mononuclear cell respiratory burst activity. AB - Opiate addiction and stress have been associated with altered immune responses. In this study, we evaluated the influence of morphine and the stress responsive opioid peptide beta-endorphin (beta-END) on O-2 and H2O2 production by cultured human peripheral blood mononuclear cells. Exposure of these cells during 48 hr of culture to morphine and beta-END at pharmacologically (10(-8) M) and physiologically (10(-12) M) relevant concentrations, respectively, markedly suppressed peripheral blood mononuclear cell O-2 and H2O2 release in response to the respiratory burst stimuli opsonized zymosan and phorbol myristate acetate. Both opioids also induced a minimal, but statistically significant, increase in resting O-2 and H2O2 generation. The modulatory effects of morphine and beta-END on peripheral blood mononuclear cell oxygen metabolism appeared to involve a classical opioid receptor, because opioid activity was blocked by naloxone and was not observed with N-acetylated-beta-END. Using purified lymphocyte and monocyte preparations, we determined that although opioids directly increase monocyte-resting oxygen metabolism, lymphocytes are the primary target cell in opioid-mediated suppression of monocyte respiratory burst activity. The release of a suppressive product from opioid-triggered lymphocytes was inhibited by cyclosporine. Based on the results of this study, we propose that opioid-mediated suppression of mononuclear phagocyte respiratory burst activity is another factor to be considered in the immunodeficiency of opiate addiction and stress. PMID- 3035016 TI - Production of platelet-activating factor by stimulated human polymorphonuclear leukocytes. Correlation of synthesis with release, functional events, and leukotriene B4 metabolism. AB - Platelet-activating factor (PAF) is a phospholipid mediator of inflammation that is synthesized by several human cell types including polymorphonuclear leukocytes (PMN). We examined the synthesis and release of PAF by stimulated human PMN under several conditions, assayed by the incorporation of [3H]acetate into PAF and by bioassay. PAF synthesis was induced by calcium ionophore A23187 (IoA), opsonized zymosan (OpsZ), and N-formyl-methionyl-leucyl-phenylalanine (FMLP) with the relative order of potency IoA much greater than OpsZ greater than FMLP. A variety of other agonists, including phorbol myristate acetate, an activator of protein kinase C and of PMN functional responses, did not stimulate PAF synthesis. PAF synthesis by PMN in response to IoA, OpsZ, and FMLP was concentration- and time dependent but release of the phospholipid was not: little PAF (1 to 10%) was released from PMN in suspension regardless of the total amount produced, the agonist, its concentration, the time of incubation, or the concentration of extracellular albumin. This was also the case with functionally altered neutrophils that had been "primed" with cytochalasin B or lipopolysaccharide or that had adhered to surfaces. PAF synthesis was tightly coupled with leukotriene B4 production by adherent PMN as well as by neutrophils in suspension, supporting the hypothesis that the two lipid autacoids may be derived from a common precursor. However, PAF synthesis could be dissociated from aggregation and surface adhesion, indicating that it is not absolutely required for these responses of activated PMN. The total amount of PAF that accumulated, but not the percentage that was released, was altered in adherent PMN compared to cells in suspension. These experiments demonstrate that PAF production and its subsequent processing by human neutrophils are highly regulated events. PAF synthesis is associated with PMN activation, but it is not a requisite for early adhesive responses of neutrophils. Because little of the PAF produced by stimulated PMN is released from the cells, it appears that PAF has an intracellular role in PMN function and/or that it may have novel intercellular effects that do not require release into the fluid phase. PMID- 3035018 TI - Reduced allorecognition of adenovirus-2 infected cells. AB - The early region 3 of adenovirus 2 encodes the membrane glycoprotein E19. This protein specifically binds class I transplantation antigens of a variety of species. Concomitant with this interaction the intracellular transport of newly synthesized class I heavy chains is abrogated. At late stages of the virus infection this leads to a notable decrease in the cell surface expression of class I antigens. We have studied how infection with adenovirus 2 influences target cell recognition by alloreactive cytolytic T lymphocytes. We found that the E19 protein-induced reduction of the HLA-B7 cell surface expression led to a greatly reduced lysis of the infected cells. These findings support our hypothesis that the E19 protein has evolved to facilitate the in vivo replication of the virus by reducing the expression of HLA class I antigens. PMID- 3035017 TI - Cross-linking membrane IgM induces production of inositol trisphosphate and inositol tetrakisphosphate in WEHI-231 B lymphoma cells. AB - The addition of anti-IgM to the immature B lymphoma cell line WEHI-231 resulted in breakdown of phosphatidylinositol 4,5-bisphosphate, generating diacylglycerol and inositol 1,4,5-trisphosphate (Ins(1,4,5)P3). These reactions have recently been demonstrated in mature resting B cells stimulated with anti-IgM, as well. In addition to Ins(1,4,5)P3, inositol tetrakisphosphate (InsP4) and inositol 1,3,4 trisphosphate (Ins(1,3,4)P3) were rapidly generated in WEHI-231 cells upon stimulation of the antigen receptor with anti-IgM. These two inositol polyphosphates are probably generated from Ins(1,4,5)P3 by phosphorylation to yield InsP4 and removal of the 5-phosphate from InsP4 to yield Ins(1,3,4)P3. It is possible that these inositol polyphosphates play a second messenger role in mediating the biologic effects of antigen-receptor signaling. It had previously been shown that anti-IgM also causes an increase in cytoplasmic free calcium. Therefore, the relationship between Ca2+ elevation and phosphoinositide breakdown was investigated. Although elevation of cytoplasmic Ca2+ with ionophores can trigger phosphoinositide breakdown, this required levels of Ca2+ well beyond those normally seen in response to anti-IgM. Thus, the Ca2+ elevation seen in response to anti-IgM cannot be the event controlling phosphoinositide breakdown. WEHI-231 cells have been shown to have a calcium storage compartment that releases Ca2+ in the presence of Ins(1,4,5)P3; therefore, it is likely that anti IgM stimulates phosphoinositide breakdown as a primary event and this leads to the elevation of cytoplasmic Ca2+. PMID- 3035019 TI - C lambda 2 and C lambda 4 immunoglobulin light chain genes in a wild-derived inbred mouse strain. AB - A genomic clone containing the lambda constant (C) region genes (C lambda 2S and C lambda 4S) has been isolated from a genomic library from the mouse strain SPE. SPE is an inbred strain derived from progenitors trapped near Grenada in Spain and has been classified as mus 3 or mus spretus. The sequence of the C lambda 2S gene is virtually identical to that of BALB/c both in the coding region and in flanking sequences, suggesting that it is an expressed gene in the SPE strain. By contrast, the C lambda 4S gene on the same cluster has diverged in sequence from that of BALB/c and contains a large deletion that precludes its normal expression. Whereas BALB/c J lambda 4 region contains substitutions that probably preclude its usage, the SPE J lambda 4 gene includes all sequences required for a functional J gene. Comparison of the C lambda 2S and C lambda 4S gene sequences with those available for BALB/c C lambda 3 and C lambda 1 confirms the close relationship between the C lambda 1-C lambda 4 and C lambda 2-C lambda 3 gene pairs. The C lambda 3 gene of BALB/c is more closely related to C lambda 2S than is C lambda 1 of BALB/c to C lambda 4S. If it is assumed that C lambda 1 and C lambda 2 are respective duplicates of C lambda 4 and C lambda 3 and that these duplications occurred at the same time, then the C lambda 2 gene has been under stronger selective pressure than C lambda 4. PMID- 3035020 TI - Maleylated-BSA suppresses IFN-gamma-mediated Ia expression in murine peritoneal macrophages. AB - Maleylated bovine serum albumin (maleyl-BSA) and other polyanionic polymers that are recognized by cell surface receptors on macrophages have been shown to induce chemotaxis, protease secretion, and tumoricidal function in this cell type. In this paper the effect of maleyl-BSA on Ia antigen expression has been evaluated. In a fashion similar to LPS, maleyl-BSA suppressed IFN-gamma-induced expression of Ia in a time- and dose-dependent manner. Also like LPS, maleyl-BSA stimulated the production and secretion of substantial amounts of PGE2 over a 24-hr period. This did not, however, appear to be the primary mechanism by which expression of Ia was suppressed, because co-treatment of the cells with indomethacin, which totally inhibited the production of PGE2, only minimally affected the suppressive activity. Surprisingly, the suppressive activity of both maleyl-BSA and LPS could be largely abrogated by co-treatment of the cells with cyclohexamide during the time period when Ia expression was sensitive to suppression. This effect was selective in that PGE2- or dibutyryl cyclic AMP-induced suppression of Ia expression was not affected by cyclohexamide treatment. The data support the concept that there are multiple molecular mechanisms involved in the negative regulation of IFN-gamma-induced Ia expression in macrophages. Such mechanisms may include, in addition to the synthesis of PGE2 and consequent elevation in intracellular levels of cyclic AMP, one or more proteins made early after treatment with either maleyl-BSA or LPS. Thus the function of some of these early gene products may be to regulate expression of functional genes such as that encoding Ia antigen. PMID- 3035021 TI - Tp44 molecules involved in antigen-independent T cell activation are expressed on human plasma cells. AB - We have analyzed cells of the B lineage for expression of the Tp44 antigen, a 44,000 homodimer detected by monoclonal antibody 9.3 on approximately 80% of mature human T lymphocytes. Previous evidence has suggested that Tp44 may function as a receptor for accessory signals in T cell activation. High level Tp44 expression was observed on plasmacytomas grown in cell culture and on plasma cells from bone marrow biopsies of multiple myeloma patients. This antigen is not present on resting B cells from either peripheral blood or lymphoid organs, or on any other B cell tumor. The growth kinetics and Ig production in plasmacytomas are not affected by the binding of antibody 9.3. Moreover, the Tp44 molecule is co-expressed with PCA-1, an antigen characteristic of plasma cells, on peripheral blood B cells stimulated in vitro to differentiate toward plasma cells. Tp44 may represent a later stage of B cell differentiation than PCA-1 because unlike the PCA-1 antigen, this molecule could not be detected on any EBV-transformed cell line or Burkitt's lymphoma lines. The m.w. of the Tp44 molecule expressed on plasma cells and on T cells is identical, as determined by immunoprecipitation of radioiodinated cell surface proteins with monoclonal antibody 9.3. This antigen might be useful in studying the mechanism of growth and differentiation of human B cells, the heterogeneity within plasma cell populations, and B cell interactions with other components of the immune system. PMID- 3035022 TI - Arachidonic acid acts as an intracellular activator of NADPH-oxidase in Fc gamma receptor-mediated superoxide generation in macrophages. AB - A protein kinase C inhibitor, 1-(5-isoquinolinesulfonyl)-2-methylpiperazine dihydrochloride (H-7), inhibited phorbol ester (12-o-tetradecanoylphorbol 13 acetate)-induced and Fc gamma receptor-mediated superoxide anion (O2-) generations in guinea pig macrophages, but the inhibitory effect on Fc gamma receptor-mediated O2- generation was only partial. Both O2- generations were inhibited extensively by a phospholipase A2 inhibitor, 4-p-bromophenacyl bromide (4-pBPB). It was confirmed in control experiments that H-7 and 4-pBPB had no direct inhibitory effect on NADPH-oxidase activity. Dose-dependent stimulation of O2- generation was induced by arachidonate in macrophages, and the arachidonate induced O2- generation was not inhibited by H-7. Arachidonate could also induce NADPH-oxidase activation in a post-nuclear fraction obtained from unstimulated macrophages and this activation was not inhibited by H-7, indicating that protein kinase C activation was not involved in this cellfree system. These results support the hypothesis that the O2- generation induced by Fc gamma receptor stimulation is mainly mediated by arachidonic acid which is released by the action of phospholipase A2 activated by receptor stimulation. Arachidonic acid seems to be acting rather directly in activating the NADPH-oxidase system of macrophage membrane. Protein kinase C may have a significant role in Fc gamma receptor-mediated O2- generation but it is not obligatory, and protein kinase C seems to activate NADPH-oxidase rather indirectly, probably by inducing the arachidonic acid release. PMID- 3035023 TI - Monoclonal antibodies against murine IFN-gamma abrogate in vivo tumor immunity against RSV-induced murine sarcomas. AB - Previous work has shown that immunization of syngeneic mice with v-src-induced sarcomas gives rise to specific protection against a lethal dose of v-src transformed fibroblasts. This immune response is mediated by Lyt-1+, Lyt-2,3+ T lymphocytes, with no apparent involvement of cytotoxic T cells, as shown in Winn type assays. Immune cells mediating tumor rejection in this system have now been further characterized, and it was found that L3T4+ T lymphocytes alone provided full protection against v-src-induced sarcomas. Moreover, the role of interferon gamma (IFN-gamma) in the tumor rejection was analyzed. A monoclonal antibody directed against this lymphokine was able to reverse the protective effect displayed by immune T lymphocytes, by eliciting highly effective T suppressor cells. It was thus concluded that T cells with L3T4 surface marker are the main thing responsible for the adoptive immunity in this tumor system, and the activity of these cells is positively modulated by lymphokines such as IFN-gamma. PMID- 3035024 TI - Isotype specificity of antigen-specific helper clone in vivo. AB - Inoculation of an immortalized clone of radiation leukemia virus (RadLV) transformed antigen (ovalbumin, OVA)-specific T cells together with the relevant carrier (OVA) into unprimed syngeneic mice results in a preferential increase in the expression of anti-OVA antibodies of the immunoglobulin (Ig)G2b and IgG2a isotypes. Identical boosting of the clone-primed mice further augments the preferential production of anti-OVA antibodies of these two isotypes. The class related helper activity is not due to nonspecific shift of class expression produced by the injected tumor cells, as a non-helper clone of RadLV-transformed T cells does not change the isotypic pattern of anti-OVA antibodies in the inoculated mice. A carrier-specific activation of the B cells is responsible for the class-restricted function of the helper clone. The isotypic profile of anti hapten antibodies in mice injected with 2,4-dinitrophenyl (DNP)-bovine serum albumin and OVA-specific helper clone is not altered. On the other hand, mice inoculated with the OVA-specific helper clone and DNP-OVA respond with a preferential elevation of anti-DNP antibodies of the IgG2a and IgG2b isotypes. The preferential class augmentation may result from carrier-specific signals delivered by the helper clone which activate B cells in vivo toward certain CH expression. Alternatively, the observed class pattern may be induced by an isotype noncommited helper clone which triggers selected population of B lymphocytes of defined differentiation status toward secretion of a restricted array of isotypes. Regardless of the mechanism of the clone-dependent class expression, the isotypic profile in most of the experiments clearly demonstrates that an antigen-specific helper clone may be one of the elements which regulates the class of antibodies to be produced in vivo under normal physiologic conditions. PMID- 3035026 TI - Labeling of the oligosaccharide moieties of immunoglobulins. PMID- 3035025 TI - Chronic experimental autoimmune myasthenia gravis induced by monoclonal antibody to acetylcholine receptor: biochemical and electrophysiologic criteria. AB - To determine whether the chronic presence of antibody to acetylcholine receptor (AChR) can account for the neuromuscular abnormalities in myasthenia gravis (MG), rats injected repeatedly with monoclonal antibody (mAb) to AChR were compared with those injected with control mAb. In a previous report, those receiving anti AChR mAb, studied ultrastructurally, had grossly simplified endplates when compared with normal controls. In this report, animals injected once or chronically for 9 to 12 wk had reduced content of muscle AChR. The chronically injected animals also had diminished miniature endplate potential amplitudes, but to a lesser extent than the reduction in AChR content. These studies establish the pathogenetic role of antibody to AChR in the induction of the ultrastructural, biochemical, and electrophysiologic hallmarks of MG. PMID- 3035027 TI - Congenital mesoblastic nephroma of infancy. PMID- 3035028 TI - Screening haemodialysis patients for infection with human immune deficiency virus (HIV). AB - A group of 484 patients having regular haemodialysis was tested for the presence of antibodies to the human immunodeficiency virus (HIV). With a commercial enzyme linked immunoassay kit, the serum of 17 appeared positive. When these 17 samples were retested by a different method, however, none was found to contain antibodies to the virus. Furthermore, evaluation of the clinical state of these 17 patients for the presence of any prodromal symptoms associated with the acquired immune deficiency syndrome was negative. It is therefore suggested that patients having regular haemodialysis are presently at low risk of contracting infection by HIV. By contrast, 81% of these patients had antibodies to cytomegalovirus. PMID- 3035029 TI - Hemocytes of the pond snail Lymnaea stagnalis generate reactive forms of oxygen. PMID- 3035030 TI - Biologic properties of LTB4 and paf-acether in vivo in human skin. AB - Using an improved skin chamber technique, the consequences of prolonged contact of leukotriene B4 (LTB4) and platelet-activating factor (paf-acether) with human dermis were evaluated quantitatively and kinetically in vivo. Leukocyte chemotaxis, histoenzymologic alterations, and modifications in vascular permeability were studied in two sets of experiments. In a first set of experiments, the dose-effect relationship of LTB4 and paf-acether on leukocyte migration was studied. LTB4 (3 X 10(-8) M to 9 X 10(-7) M) in Hanks' balanced salt solution (HBSS) elicited an intense dose-dependent and time-dependent neutrophil migration. Paf-acether, at the same concentration range, induced a significant increase in cell migration only at 9 X 10(-7) M and when diluted in HBSS containing 0.25% serum albumin (HBSS-BSA). Histoenzymologic analysis demonstrated that LTB4 in vivo induced degranulation of most of the neutrophils migrating through the dermis. Paf-acether caused mild degranulation of neutrophils and induced the appearance of degranulated basophils in dermal vessels. A second set of experiments was designed to study simultaneously the modifications in vascular permeability and cell migration induced by LTB4 and paf acether, with or without prostaglandin E2 (both at a concentration of 3 X 10(-7) M in HBSS). Since spontaneous protein diffusion in HBSS progressively declined up to a plateau reached after 20 h (1.2 +/- 0.15 mg of proteins/cm2/2 h), these experiments were carried out after a 20-h equilibration period. Leukotriene B4 induced a late and slight increase in vascular permeability. Paf-acether did so intensely and transiently. Prostaglandin E2 significantly enhanced protein diffusion and neutrophil migration induced by LTB4 and, to a lesser extent, by paf-acether. Interestingly, despite the reintroduction into the skin chambers of freshly prepared solutions containing the mediators, leukocyte migration and protein diffusion progressively decreased during the experiments. This suggests the local production of anti-inflammatory factors that inhibit local mediators and thus regulate the inflammatory response. PMID- 3035031 TI - Oxygen intermediates are involved in ultraviolet radiation-induced damage of Langerhans cells. AB - Experiments were conducted to determine whether ultraviolet (UV) radiation exerts its effect through the generation of oxygen intermediates on Langerhans cells (LC). Guinea pigs were exposed to one single dose of UVB (0.9-2.7J/cm2), and biopsy specimens were taken 5 days after the irradiation. The population of LC was evaluated using ATPase-stained epidermal sheets. These exposures reduced the number of LC to 20-25% of the original density. On the other hand, superoxide dismutase (SOD) (0.02-0.2 mg), a scavenger of superoxide anion, which had been injected intradermally just before UV radiation, significantly prevented the depletion of LC, although not completely (37-40% of the original density). The injection immediately after the exposure was still significantly effective, but less so. Other scavengers of oxygen intermediates including catalase, D-mannitol, and L-histidine revealed no detectable effect. A single exposure of UVB at doses of 0.3-0.6 J/cm2 did not deplete the ATPase-positive LC. However, the same dose of UVB reduced the number of LC to 70%, when exposed after the injection of an SOD inactivator, diethyldithiocarbamate, possibly due to inactivation of physiologically existing SOD. These observations indicate that oxygen intermediates such as superoxide anion or its subsequent species are generated by UV radiation exposure and damage the epidermal LC. PMID- 3035033 TI - The antiviral activity of recombinant human tumor necrosis factor-alpha. PMID- 3035032 TI - Behavior of epidermolysis bullosa fibroblasts in a hydrated collagen lattice. AB - To determine if an altered ability to contract a hydrated collagen lattice is characteristic of fibroblasts from patients with recessive dystrophic epidermolysis bullosa (RDEB), we examined contraction by fibroblasts from normal subjects and patients with RDEB, dominant dystrophic epidermolysis bullosa (DDEB), and dominant epidermolysis bullosa simplex (DEBS). An extremely broad range of contractility (normal, poor, and hypercontraction) was observed in all types of epidermolysis bullosa (EB). When contraction in control fibroblasts was defined as the mean +/- 2 SD, (all control values were within this range) and the data were analyzed by the chi-square test, only 32% of EB cells fell within this range, with 47% poorly contractile and 21% hypercontractile. These data, derived from 34 patients, indicate that no single genetic defect resulting in altered contractility in the 3 distinct types of EB is likely. Neither cell viability, collagenase expression, nor PGE2 synthesis as correlated with gel contraction in any group. Indomethacin had no effect on contraction in RDEB. It is possible that the genetic defects in EB cause blister formation in vivo and may lead in some way to an abnormal interaction of fibroblasts with the extracellular matrix resulting in an altered collagen lattice contraction in vitro. PMID- 3035034 TI - Time course of interferon levels, antiviral state, 2',5'-oligoadenylate synthetase and side effects in healthy men. AB - The kinetics of the biologic response following a single intramuscular injection of 18 X 10(6) units of recombinant human interferon-alpha 2a (rHuIFN-alpha 2a) was investigated during 11 courses in 10 healthy individuals. Serial peripheral blood mononuclear cell (PBM) samples were assayed for their biologic responsiveness to rHuIFN-alpha 2a by measuring both their 2',5'-oligoadenylate (2 5A) synthetase activity and their resistance to in vitro vesicular stomatitis virus (VSV) infection. A significant increase in 2-5A synthetase levels occurred at 6 h, and enzyme levels returned to baseline values between 96 and 104 h postinjection. Protection of PBMs from VSV infectivity began within 1 h and lasted up to 144 h postinjection. The clinical side effects induced by IFN administration and serum IFN levels were not parallel over time with the antiviral effects observed. This study defines the time course of the biologic response induced by rHuIFN-alpha 2a in healthy volunteers. A parallel time course between the induction of 2-5A synthetase activity and the development of the antiviral state in PBMs was demonstrated. PMID- 3035035 TI - [Operative cases of intrathoracic malignant fibrous histiocytomas]. PMID- 3035036 TI - [Biochemical study on the thyrotropic activity in urinary immunoreactive-human chorionic gonadotropin of patients with trophoblastic disease]. AB - We have studied the biochemical properties of urinary immunoreactive-human Chorionic Gonadotropin (IR-hCG) using the human thyroid gland in vitro slice method to investigate the thyrotropic activity in purified IR-hCG of patients with thyroid hyperfunction associated with trophoblastic disease. IR-hCG was extracted using a kaolin-acetone-alcohol concentration and purified by DEAE Cellulose, Sephadex G-100 and DEAE Sephacel column chromatographies according to Nishimura et al. with slight modifications. The substance with thyrotropic activity (hCT) was found in 3 cases of trophoblastic disease with hyperthyroidism among the 14 cases investigated. HCT consists of two dissimilar subunits, namely, hCT alpha-subunit and hCT beta-subunit like hCG, whose molecular weights are estimated at about 15,000 and 20,000 respectively and have carbohydrate moiety bound to concanavalin A. The thyrotropic activity was shown by the recombinant hybrid molecules, hCT beta-subunit and hCG alpha-subunit, whereas no potency was found in hCT beta-subunit only. The carboxyterminal residues of hCT (Ile-Leu-Pro Glu) were not digested by carboxypeptidases Band Y. These results suggest that hCT is an intrinsic activity of hCG and cross-reacts immunologically, while the amino acid residues of the carboxyterminus of the hCT beta-subunit are different from those of the hCG beta-subunit. PMID- 3035037 TI - Relationship between atrial natriuretic polypeptide and cyclic 3'5'-guanosine monophosphate in human plasma. AB - To examine the interrelationship between human atrial natriuretic polypeptide (hANP) and cyclic 3'5'-guanosine monophosphate (cyclic GMP), plasma concentrations of these compounds were determined in 61 disease-free humans, as controls, and in 35 patients with congestive heart failure. Levels of plasma hANP (199.6 +/- 53.7 pg/ml) and cyclic GMP (12.6 +/- 1.7 pmol/ml) in patients with congestive heart failure were significantly higher than in the control subjects (hANP 57.1 +/- 2.8 pg/ml, cyclic GMP 5.2 +/- 0.3 pmol/ml). Although plasma hANP concentrations in the patients with congestive heart failure tended to increase with the severity of cardiac dysfunction, there was no significant correlation between the levels of plasma hANP and the grade of heart failure, classified according to the New York Heart Association. However, a significant correlation was found between plasma hANP and cyclic GMP concentrations in both the healthy subjects and the patients with congestive heart failure, and a weak positive correlation between plasma hANP and cyclic 3'5'-adenosine monophosphate (cyclic AMP) concentration in the patients with congestive heart failure. Thus, changes in plasma cyclic GMP concentration depend to some extent on the plasma concentrations of hANP. PMID- 3035038 TI - Parapharyngeal malignant ectomesenchymoma: combined malignant fibrous histiocytoma and primitive neuroectodermal tumour with neuroglial differentiation. AB - A parapharyngeal ectomesenchymoma consisting of mixed malignant fibrous histiocytoma and primitive neuroectodermal tumour with neuroglial differentiation occurred in a 36-year-old woman. Immunohistochemical and electron microscopic studies verified the combined mesenchymal and neuroectodermal components within the tumour. Only 9 similar cases, including the present one, are on record. The patients ranged in age from 6 months to 49 years, with an average age of 18 years. The male:female ratio was 4:5. The tumour location was widespread. All reported ectomesenchymomas were histopathologically malignant. Recurrence and/or metastasis was/were common. The neural crest has been suggested as the origin of the ectomesenchymomas. PMID- 3035040 TI - Evidence of abnormal dopaminergic control of prolactin in patients with hypothalamic and pituitary tumors. AB - Prolactin secretion was investigated in an attempt to identify the patterns of responses in different types of tumors. Forty four patients were studied: thirty patients with prolactinomas (Group 2); nine patients with growth-hormone (GH) adrenocorticotropic hormone (ACTH)-secreting pituitary tumors and hypothalamic tumors (Group 3); and five patients with non-secreting pituitary tumors (Group 4). A control group (Group 1) consisted of 60 healthy subjects (30 males and 30 females). All were submitted to testing by nomifensine (Nom), domperidone (Dom) and thyrotropin releasing hormone (TRH). The prolactin levels were measured by radioimmunoassay (RIA). In group 2 the suppression of PRL with Nom and the stimulation with Dom and TRH were significantly lower than in the control group (p less than 0.001). There was no statistically significant difference between groups 2 and 3 in the suppression with Nom. The increase with Dom in group 3 was significantly greater than that in group 2 (p less than 0.001) and less than that in the control group (p less than 0.005). The rise in PRL with TRH was also significantly higher in group 3 than in group 2 (p less than 0.001) and similar to that of the control group. Group 4 gave the same results as the control group to all 3 tests. Our results indicate a dopaminergic irregularity in the hypothalamic and GH-ACTH-secreting pituitary tumors, thus supporting a hypothalamic etiopathogenesis of these tumors. The normality of the GH-ACTH secreting pituitary tumors and hypothalamic tumor responses to TRH is one more factor in differentiating these from prolactinomas. The normal response of the non-secreting tumors may involve a primary pituitary etiology of these tumors. PMID- 3035039 TI - Secretion of lipids, apolipoproteins, and lipoproteins by human hepatoma cell line, HepG2: effects of oleic acid and insulin. AB - The aim of this study was to determine the effect of oleic acid and insulin on the secretion of lipoproteins by HepG2 cells grown in minimum essential medium. Triglycerides were the major neutral lipid (57% of total) and apoB was the predominant apolipoprotein (56% of total) secreted by these cells. The addition of oleate resulted in a two-fold increase in the concentration of neutral lipids but only a slight to moderate increase in the apolipoprotein (A-I, A-II, B, and E) levels. The secretion of very low density lipoproteins (VLDL) was stimulated by 425%, low density lipoproteins (LDL) by 77%, and high density lipoproteins (HDL) by 68%. Whereas neutral lipid composition of LDL was unchanged, the VLDL particles contained a significantly higher percentage of triglyceride and lower percentages of cholesterol and cholesteryl esters compared with VLDL secreted in the absence of oleate. Oleate had no significant effect on the composition of apolipoproteins in VLDL, LDL and HDL. In basal medium, insulin caused a significant decrease in the secretion of neutral lipids and apolipoproteins, particularly triglycerides and apoB. In addition to a 60-68% reduction in the total concentration of VLDL and LDL, insulin altered their composition by producing particles that had a significantly lower content of triglycerides, contained less apoB, and were deficient in apoE. There were no major changes in the concentration or composition of HDL particles. Insulin had a similar but less pronounced effect on the concentration and composition of lipoproteins secreted in the presence of oleate. The increased accumulation of triglycerides in the HepG2 cells concomitant with their reduced levels in the medium suggests that insulin may affect the secretion rather than synthesis of triglyceride-rich lipoproteins. PMID- 3035041 TI - Narcotics: the latest chapter in a long story. A review. AB - Extraordinary significant discoveries have been made in the pharmacology of opioid drugs in the last fifteen years. One has to go back to the beginning of the eighteenth century to find a comparably significant period. The advances in our understandings and uses of opioids have taken place in four different areas: postulating the existence of several types of receptors and the discovery of receptors sites, the isolation of endogenous opioids, the development of new opioid substances, and the use of new routes and modes of administration of narcotics. PMID- 3035042 TI - Comparison of biophysical parameters of primary low and high metastatic Lewis lung tumor cells. AB - Lewis lung tumor cells from tumors of both high and low metastatic potential were isolated from the leg muscle of C57-B1 mice and were purified by density gradient centrifugation. The purified cells retained most of their tumor-forming capacity; however, the difference in the metastatic capacity of the 2 cell lines increased with the purification process. The purified and viable cells were subjected to different biophysical studies. Electron spin resonance studies showed that the motional freedom of lipid and protein molecules of the cellular membrane is higher in the low metastatic cell population. Both cell types reduce penetrating and non-penetrating free radicals, but the reducing ability of the low metastatic cell population is 4 times greater than that of the high metastatic cell line. These findings argue for increased membrane dynamics in the low metastatic cell population. PMID- 3035043 TI - Effect of Salmonella typhimurium porins on the cardiovascular and renal apparatus. AB - The cardiovascular effects of porins were evaluated using porins isolated from Salmonella typhimurium SH5014. In dogs porins depress arterial systemic pressure, vasomotor reactivity of norepinephrine and peripheral vagal stimulation. They are capable of modifying the sinocarotidal baroreceptor reactivity. In mice porins increase the cardiotoxic effects of isoprenaline, thyroxine, emetine and of p nitrophenol. In rats porins increase the arrhythmogenic and lethal effects of BaCl2 and also give rise to renal lesions, probably at the tubular level. PMID- 3035045 TI - Measurement of circulating corticotrophin-releasing factor in man. AB - A radioimmunoassay was developed to measure corticotrophin-releasing factor (CRF 41) extracted from human plasma using Vycor glass. Assay sensitivity was 20 ng/l and intra- and interassay coefficients of variation were 10.2 and 11.4% respectively. The normal range of plasma CRF-41 was less than 20-110 ng/l (n = 46). Plasma concentrations of CRF-41 in patients with Cushing's disease. Nelson's syndrome and Addison's disease were within the normal range. No correlation was found between CRF-41 and ACTH in these syndromes. Two patients with the ectopic ACTH syndrome had increased plasma concentrations of CRF-41. In normal subjects no changes in plasma CRF-41 occurred after insulin-induced hypoglycaemia, treatment with dexamethasone or feeding, and changes in the concentrations of CRF 41 did not reflect circadian changes in plasma concentrations of cortisol. Concentrations of immunoreactive CRF in plasma of women in the third trimester of pregnancy were increased (550-9300 ng/l) and gel filtration chromatography showed that this comprised CRF-41 and a higher molecular weight form. Reversed-phase high-performance liquid chromatography also revealed multiple peaks of immunoreactive CRF in extracts of plasma and placenta. PMID- 3035044 TI - Significant enhanced superoxide anion (O2-) production in vitro by peripheral blood monocytes of lepromatous leprosy patients stimulated with liposome and suppression by C-reactive protein (CRP). AB - Peripheral blood monocytes (PBM) from normal individuals, infectious mononucleosis (IM) and leprosy patients were stimulated with liposome. The mean and standard error of superoxide anion (O2-) generated in nm/1.5 X 10(5) PBM/well for 5 normal subjects and 3 IM patients was 2.9 +/- 0.5 and 3.1 +/- 0.2, respectively. Monocytes stimulated with 100 ng C-reactive protein (CRP) incorporated into liposome gave values of 3.3 +/- 0.3 and 2.7 +/- 0.1 nm O-2/1.5 X 10(5) PBM/well for normals and IM, respectively. No significant differences in O2- production between liposome and liposome with incorporated CRP were shown. PBM from lepromatous patients demonstrated a significant (p less than 0.01) increase in O2- production with liposome alone compared with tuberculoid patients (3.5 +/- 0.4 vs 1.8 +/- 0.3). The most dramatic suppression of O2- shown when purified CRP was added to the mixtures in all groups examined [0.4 +/- 0.1 (500 ng), 0.3 +/- 0.0 (500 ng), 1.5 +/- 0.1 (100 ng), and 1.3 +/- 0.6 (100 ng) nm O2 /1.5 X 10(5) PBM/well for normals, IM, lepromatous, and tuberculoid, respectively]. Results of O2- formation with incorporation of CRP into liposome as compared with liposome alone had no significant effect on PBM of lepromatous or tuberculoid patients. It is suggested that CRP may play a significant role in regulation of oxygen free radicals formed during acute and chronic inflammatory episides. PMID- 3035047 TI - Inhibition of FSH-stimulated granulosa cell function by a synthetic fragment of the porcine inhibin alpha-subunit: evidence for involvement of GnRH receptors. AB - The bioactivity of a synthetic peptide fragment which mimics the N-terminal sequence of the 134-amino-acid porcine inhibin alpha-subunit (pl-alpha 1-26 Gly27Tyr28-OH) was tested and compared with the bioactivity of GnRH in rat granulosa cell cultures. Granulosa cells from immature female rat ovaries were cultured with hFSH and testosterone to stimulate the production of cyclic AMP, progesterone and oestradiol. Addition of pl-alpha 1-26-Gly27Tyr28-OH to the culture medium caused a dose-dependent suppression of all three parameters (ID50 700-1,000 nmol/l). GnRH caused similar but higher-potency inhibition (ID50 2-4 nmol/l). Suppression of granulosa cell function by both peptides was fully reversible by a synthetic GnRH antagonist. Moreover, specific binding of the porcine inhibin fragment to ovarian GnRH receptors was demonstrated by radioreceptor assay. This is evidence that the porcine inhibin alpha-subunit fragment suppresses FSH-induced rat granulosa cell function via a mechanism of action similar to that of GnRH. PMID- 3035049 TI - Role of potassium in vasopressin-induced production of cyclic AMP in rat renal papillary collecting tubule cells in culture. AB - The effect of potassium (K)-free medium on the stimulation of cyclic AMP (cAMP) production by arginine vasopressin (AVP) and forskolin was examined in rat renal papillary collecting tubule cells in culture. All experiments were performed in the presence of 3-isobutyl-l-methylxanthine (0.5 mmol/l). Cellular cAMP levels in response to 1 nmol and 0.1 mumol AVP/1 were 430.9 +/- 42.1 (S.E.M.) and 501.8 +/- 43.6 fmol/micrograms protein per 10 min respectively; these levels were significantly (P less than 0.01) higher than those in the vehicle-treated group (126.6 +/- 23.3 fmol/micrograms protein per 10 min). The cellular cAMP response to 1 nmol AVP/1 was significantly attenuated after 24 and 72 h of exposure of cells to K-free medium, cellular concentrations of cAMP being 280.2 +/- 37.1 and 233.0 +/- 9.6 fmol/microgram protein per 10 min respectively. The response of cAMP to AVP remained unchanged when the cells were preincubated with K-free medium for 1 h. Similarly, forskolin (20 nmol/l)-stimulated cellular cAMP production was also significantly impaired after 24 or 72 h of exposure of cells to K-free medium. When the cells preincubated in K-free medium were again exposed for 1 h to K-replete medium containing 5 mmol KC1/1, cellular cAMP production in response to AVP or forskolin recovered totally. Cellular protein and ATP content and cellular viability were not altered by exposure of cells to K-free medium for 24 h, and thus the impaired cAMP response to AVP or forskolin in the K-depleted cells was independent of altered cellular viability and source of ATP.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3035048 TI - Autoradiographic localization of binding sites for oxytocin and vasopressin in the rat kidney. AB - The distribution of [3H]vasopressin- and [3H]oxytocin-binding sites was examined, using an autoradiographical technique, in the kidney of Long-Evans and Brattleboro rats. Two types of binding sites with affinities in the nanomolar range were detected: one, located on glomeruli, bound both vasopressin and oxytocin; the other, on collecting ducts, bound vasopressin selectively. In the presence of 10 mumol oxytocin/l, [3H]vasopressin labelling was abolished in glomeruli, but only reduced in collecting ducts; [3H]oxytocin labelling was completely abolished by 10 mumol vasopressin/l. These observations are discussed in relation to known effects of neurohypophysial hormones on renal physiology. PMID- 3035046 TI - A phaeochromocytoma presenting with Cushing's syndrome associated with increased concentrations of circulating corticotrophin-releasing factor. AB - The case is described of a 61-year-old male who presented with hypertension and Cushing's syndrome which resolved on excision of a unilateral adrenal mass. Histology of the tumour revealed a benign phaeochromocytoma which immunostained for corticotrophin-releasing factor (CRF-41) but not for ACTH. Preoperative plasma concentrations of immunoreactive CRF-41 were increased, and gradients for both CRF-41 and ACTH were demonstrated across the tumour. Post-operatively, CRF 41 was undetectable in plasma. The tumour contained high concentrations of immunoreactive CRF-41 which co-eluted with synthetic human CRF-41 on reversed phase high-performance liquid chromatography. Tumour CRF-41 stimulated the release of ACTH in a dose-dependent manner from isolated rat anterior pituitary cells. We conclude that this tumour secreted CRF-41 and ACTH and had the capacity to produce ACTH-dependent Cushing's syndrome directly by secreting ACTH and indirectly by secreting CRF-41 to stimulate ACTH secretion from the anterior pituitary. PMID- 3035050 TI - Calmodulin inhibitors and 8-(N,N-diethylamino)-octyl-3,4,5-trimethoxybenzoate do not prevent the inhibitory effect of prostaglandin F2 alpha on cyclic AMP production in isolated rat corpora lutea. AB - In the rat corpus luteum, prostaglandin F2 alpha (PGF2 alpha) rapidly inhibits LH induced cyclic AMP (cAMP) production when given in vivo or to isolated corpora lutea, but not to broken-cell preparations. The suggestion that increased cytosolic calcium concentration mediates PGF2 alpha action was investigated in corpora lutea of pseudopregnancy induced in immature rats by administration of pregnant mare serum gonadotrophin (15 i.u.). Isolated 10-day-old corpora lutea were incubated for 90 min with LH (5 micrograms/ml), PGF2 alpha (10 mumol/l) and other additions, and cAMP concentration in the tissue was estimated. The putative inhibitor of intracellular calcium release or action, 8-(n,N-diethylamino)-octyl 3,4,5-trimethoxybenzoate (TMB-8; 30 or 150 mumol/l), did not abolish the effect of PGF2 alpha. Similarly ineffective was the combination of TMB-8 (150 mumol/l) and calcium-depleted medium (free ionized calcium concentration, 30 nmol/l). Calmodulin inhibitors of three different chemical structures were then tested. The phenothiazine trifluoperazine, at 300 as well as 30 mumol/l, did not interfere with the inhibitory effect of PGF2 alpha on cAMP, while suppressing (at 300 mumol/l) progesterone secretion in LH-treated tissue. Furthermore, inhibition by PGF2 alpha was not impaired by pimozide, a diphenylbutylpiperidine (25 and 50 mumol/l) nor by N-(6-aminohexyl)-5-chloro-1-naphthalene sulphonamide (W-7; 15 and 45 mumol/l). In the presence of LH alone, W-7 (45 mumol/l) inhibited and TMB-8 (30 mumol/l augmented cAMP accumulation, indicating that the luteal tissue was effectively exposed to these compounds. Thus, drugs known to inhibit calcium- and calmodulin-dependent processes in a variety of tissues did not abolish the inhibitory action of PGF2 alpha on luteal cAMP production. PMID- 3035051 TI - Recovery of the components of the hypothalamo-pituitary-adrenocortical axis in the rat after chronic treatment with prednisolone. AB - Male Wistar-derived rats (200-250 g) were treated for 14 days with prednisolone 21-sodium succinate at a concentration of 1035 mumol/l in their drinking water. The drug was then replaced with normal tap water and groups of animals were killed at various times during recovery, trunk blood being collected after decapitation. At the same time, hypothalamic slices, anterior pituitary gland fragments and adrenals were removed and their responsiveness assessed by exposure to appropriate stimuli in vitro. Tissues were also extracted to measure changes in content of hormones during recovery. Treatment with prednisolone produced marked reductions in body weight gain, adrenal weight and pituitary ACTH content, but no significant change in hypothalamic corticotrophin-releasing factor (CRF) bio- or immunoreactivity. The ACTH content was restored by 5 days after withdrawal but adrenal weight remained significantly reduced after 9 days of recovery. The responsiveness of the hypothalamus to acetylcholine in vitro was markedly inhibited and was still significantly reduced 7 days after withdrawal. The responsiveness of the anterior pituitary gland to synthetic CRF or arginine vasopressin and that of the adrenal gland to ACTH added in vitro were restored simultaneously after 7 days of withdrawal. In vivo, recovery was assessed by measurement of the response to laparotomy stress. Treatment with prednisolone prevented the increase in the plasma concentrations of ACTH and corticosterone produced by stress, and these responses recovered by 5 days (corticosterone) and 7 days (ACTH) after withdrawal. The abolition of the circadian rhythms of ACTH and corticosterone by treatment was also reversed by 5 days after withdrawal.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3035052 TI - Leukotriene release by endometrium and myometrium throughout the menstrual cycle in dysmenorrhoea and menorrhagia. AB - Endometrium and myometrium were collected at hysterectomy from 21 women with measured menstrual blood loss. Eight women complained of dysmenorrhea and the remaining 13 had pain-free periods. Specimens were obtained throughout the menstrual cycle (menstrual, n = 5; follicular, n = 4; early luteal, n = 3; mid luteal, n = 5; late luteal, n = 4). Leukotriene C4, leukotriene D4 and leukotriene E4 release were examined using a short-term incubation technique. Endometrial leukotriene release, which was always significantly greater than myometrial release, changed throughout the menstrual cycle and the highest concentrations were found during menstruation. Endometrial, but not myometrial, leukotriene concentrations were significantly higher in tissues obtained from women with a complaint of dysmenorrhoea compared with those in tissue from pain free women. No correlation was found between leukotriene release in either endometrium or myometrium and menstrual blood loss (range 15-457 ml). PMID- 3035053 TI - Specific binding sites for gonadotrophin-releasing hormone, LH/chorionic gonadotrophin, low-density lipoprotein, prolactin and FSH in homogenates of human corpus luteum. II: Concentrations throughout the luteal phase of the menstrual cycle and early pregnancy. AB - Corpora lutea were obtained from 52 women undergoing laparotomy during the luteal phase of the menstrual cycle. In addition, stromal, thecal and granulosa cell preparations were obtained from seven women undergoing ovariectomy during the late follicular-preovulatory phase of the cycle. The specific binding of a 125I labelled gonadotrophin-releasing hormone (GnRH) agonist [D-Ser(But)6] GnRH(1-9) ethylamide (buserelin) and of human chorionic gonadotrophin (hCG), human FSH (hFSH), human prolactin (hPRL) and human low-density lipoprotein (hLDL) to tissue homogenates was measured under optimal conditions. Bound LH/hCG was estimated by elution with acid-citrate buffer, followed by radioimmunoassay of released hormone. Binding of GnRH agonist, though variable, was highest in mid-luteal corpus luteum and high binding was also present in three out of four corpora lutea of pregnancy. Binding of LH/hCG increased significantly with luteinization, reaching maximal levels in the mid-luteal phase before falling significantly. Occupancy of LH receptors by bound LH was relatively constant throughout the luteal phase (10.7-35.3%), but occupancy increased to greater than 90% in corpora lutea from early pregnancy. Binding of hFSH was variable, with only five out of 50 corpora lutea having binding greater than 10 pg/micrograms DNA. Similarly, hPRL binding varied markedly with only six out of 44 having binding greater than 50 pg/micrograms DNA. Binding of LDL was highest in the early- to mid-luteal phases of the cycle. In corpora lutea from all stages of the menstrual cycle (excluding corpora albicantia), GnRH agonist binding was highly correlated with the levels of unoccupied and occupied LH receptors (P less than 0.001; n = 49 and n = 48 respectively) and with LDL receptors (P less than 0.002; n = 49). Binding of GnRH agonist was also correlated with PRL binding (P less than 0.05; n = 21) but not with FSH receptors (P greater than 0.4; n = 25). In addition, LDL binding was associated with PRL (P less than 0.005; n = 21) and FSH receptors (P less than 0.05; n = 25) and with endogenously bound LH (P less than 0.03; n = 48), but not with unoccupied LH receptors (P = 0.8; n = 49). Moreover, in corpora lutea from the mid-luteal phase, there was a strong association between GnRH agonist binding and LDL receptors (P less than 0.02; n = 23). The correlations between GnRH agonist binding and a number of important indices of luteal function suggest a physiological role for GnRH-like factors in the human corpus luteum. PMID- 3035054 TI - Determining the presence and origin of collagenase in human periapical lesions. PMID- 3035055 TI - A local interneurone which receives differential input from the medial and lateral giant axons. PMID- 3035056 TI - Active lambda and kappa antibody gene rearrangement in Abelson murine leukemia virus-transformed pre-B cell lines. AB - The two Abelson murine leukemia virus (A-MuLV)-transformed cell lines, BM18-4 and ABC-1, undergo immunoglobulin L-chain gene recombination during passage in tissue culture. BM18-4 cells are capable of kappa gene recombination, whereas ABC-1 cells are capable of both kappa and lambda gene recombination. The expression of H chains is apparently not necessary for continuing L chain gene recombination in either of these cells, although H-chain expression may have been involved in the initiation of L-chain gene recombination. All ABC-1 cells that have lambda gene rearrangements also display recombined kappa alleles, supporting the hypothesis that kappa and lambda gene recombination are initiated in an ordered, developmentally regulated manner in maturing B cells. However, analyses of the ABC-1 line indicate that pre-B cells that have initiated lambda gene recombination do not terminate kappa gene rearrangement. The lambda gene recombinations that occur in the ABC-1 cell line indicate that the germline order of lambda gene segments is: 5' ... V lambda 2 ... J lambda 2C lambda 2-J lambda 4C lambda 4 ... V lambda 1 ... J lambda 3C lambda 3-J lambda 1C lambda 1 ... 3'. In addition, the frequencies of lambda 1, lambda 2, and lambda 3 gene recombinations among ABC-1 cells are quite different than the frequencies of B cells producing lambda 1, lambda 2, and lambda 3 L-chains in the mouse. RS DNA recombinations also occur in the BM18-4 and ABC-1 cell lines, supporting the notion that Ig gene recombinases are involved in RS rearrangement. Recombined RS segments are infrequent among BM 18-4 cells but common among ABC-1 cells, suggesting that RS recombinational events often occur in maturing pre-B cells just before initiation of lambda gene rearrangements. This developmental timing is consistent with the hypothesis that RS recombination may be involved in the initiation of lambda gene assembly. PMID- 3035057 TI - Functional T lymphocytes are required for a murine retrovirus-induced immunodeficiency disease (MAIDS). AB - Athymic nu/nu mice were found to be resistant to the immunodeficiency disease and lethality induced in normal mice by the injection of the LP-BM5 mixture of murine retroviruses (LP-BM5 MuLV). Susceptibility to disease induction was reconstituted by injection of nu/nu mice with purified, mature T lymphocytes. The extent of viral replication of both the ecotropic and mink cell focus forming (MCF) components of LP-BM5 MuLV was equivalent in both nu/nu and normal animals. Retrovirally-induced immunodeficiency disease in mice (MAIDS) is thus dependent upon the presence of functional T lymphocytes, and high virus titers in athymic mice have little or no effect on the immune system. PMID- 3035058 TI - Immunofluorescent localization of the Ca2+-dependent proteinase and its inhibitor in tissues of Crotalus atrox. AB - The native 108,000 dalton Ca2+-dependent proteinase (CDP) and its 115,000 dalton protein inhibitor (CDPI) were purified from bovine skeletal muscle using native polyacrylamide gel electrophoresis and were used to elicit antibody production in rabbits and BALB/c mice. Polyclonal antibodies were purified as IgG fractions by column chromatography; monoclonal antibodies were produced by the hybridoma technique. Indirect immunofluorescence localization of CDP and CDPI in tissues of Crotalus atrox show both proteins to be ubiquitous. Both occur in the cytoplasm and are absent from the cell membrane and the nucleus; CDPI is also present in the I-band of skeletal muscle. PMID- 3035059 TI - Fine structure of cells infected with human cytomegalovirus after treatment with 9-(1,3-dihydroxy-2-propoxymethyl)guanine. AB - Infection with human cytomegalovirus (CMV) is characterized by cytological changes which are readily visualized by electron microscopy using ultrathin sections of infected cells. Treatment of such cells with 9-(1,3-dihydroxy-2 propoxymethyl)guanine (DHPG), a potent inhibitor of CMV, is effective when initiated at early or late times after infection and the response to such treatment has been studied by fine structural analysis. Inhibition of viral DNA synthesis by DHPG treatment (50 microM) late in virus infection resulted in a cessation of virus growth accompanied by a lack of development and possible regression in skein-like intranuclear inclusions together with a depletion in cytoplasmic dense bodies. Such changes were accompanied by the appearance of nuclear dense bodies. These were also present when virus growth was reduced (5 microM-DHPG) rather than completely inhibited (50 microM-DHPG) by treatment initiated from the time of infection. The nuclear bodies were predominantly of a reticular type structure after the early treatment but mainly of a homogeneous form when virus growth was interrupted at late times. Their presence appeared to be connected with the ability of infected cells to initiate the synthesis of late proteins and their morphology may relate to the extent of such protein synthesis. Unlike cytoplasmic dense bodies, provisional findings on the characterization of the nuclear bodies suggested that the 69K matrix protein was not present in abundance. PMID- 3035061 TI - Characterization of bovine viral diarrhoea-mucosal disease virus-specific proteins in bovine cells. AB - The presence of virus-specific polypeptides in bovine viral diarrhoea-mucosal disease (BVD) virus-infected bovine cells was studied by radiolabelling in the presence of a hypertonic initiation block (HIB) and by analysis by SDS-PAGE. These experiments were complemented by radioimmunoprecipitations with anti-BVD hyperimmune serum of infected cells labelled under isotonic conditions. A total of 12 polypeptides (Mr 165, 135, 118, 80, 75, 62, 56 to 58, 48, 37, 32, 35 and 19, all X 10(-3)) were identified in infected cells. Time course analysis of the induction of the viral polypeptides indicated that they could be detected as early as 4 h post-infection and their synthesis reached a plateau between 12 and 20 h post-infection. The most abundant polypeptides were the ones that could be detected earliest. HIB was found to be an excellent adjunct to existing techniques in the identification of viral polypeptides. Seven of these polypeptides had not been reported previously. This is the first report of the direct detection of BVD virus-induced polypeptides in infected cells without the aid of immunoprecipitation. The sum of the molecular masses of these polypeptides is greater than the coding capacity of the genome; therefore precursor-product relationships must exist between these polypeptides. PMID- 3035060 TI - Synthesis of cytomegalovirus DNA is an antiviral target late in virus growth. AB - The mechanism of action of 9-(1,3-dihydroxypropoxymethyl)guanine (DHPG) and phosphonoformic acid (PFA) but not 5-fluorouridinedeoxyribose (FUdR), provides selective action against cytomegalovirus (CMV)-coded events and this was used to demonstrate that the synthesis of viral DNA was continuous during the extended phase of virus growth. The synthesis de novo of viral DNA was measured by restriction enzyme analysis after exposure to [32P]orthophosphate and its interruption by DHPG or PFA resulted in a cessation in the extrusion of infective virus from treated cells. The rate of decline in infectivity appeared to correspond to the failure of cells to maintain the synthesis of late proteins once DNA synthesis was blocked. Thus, regulation of late protein synthesis appeared to be linked to synthesis de novo of viral DNA even at late stages in CMV growth. The synthesis of the polyamines spermidine and spermine, considered obligatory for CMV growth, was unaffected by early or late inhibition of viral DNA and this showed that some virus-induced events were unaffected by the restriction on virus growth by DHPG. This provided evidence that polyamine biosynthesis was a target independent of viral DNA synthesis per se, which may be important in future considerations of combined drug therapies. PMID- 3035062 TI - Glycoproteins of bovine viral diarrhoea-mucosal disease virus in infected bovine cells. AB - Bovine cell cultures infected with bovine viral diarrhoea-mucosal disease (BVD) virus were radiolabelled with L-[35S]methionine or D-[2-3H]mannose followed by analysis of the labelled polypeptides by radioimmunoprecipitation and polyacrylamide gel electrophoresis in one and two dimensions. Six glycoproteins were detected in infected cells. Two abundant species had Mr of 48K and 56K to 58K while the less abundant species had Mr of 118K, 75K, 65K and 25K. When cells were radiolabelled with L-[35S]methionine in the presence of tunicamycin 56K to 58K migrated with apparent masses of 54K (a minor species) and 48K to 50K (the major molecular species) in PAGE. Endoglycosidase F digestion of virus-induced polypeptides caused a 4K to 6K reduction in the apparent molecular mass of 56K to 58K yielding a single 52K digested product, indicating that the heterogeneity of 56K to 58K was due to differences in the oligosaccharide moieties. Tunicamycin caused a drastic reduction in the yield of infectious virus indicating that the carbohydrate moieties serve a critical role in the infectious cycle of BVD virus. PMID- 3035063 TI - Expression of bluetongue virus group-specific antigen VP3 in insect cells by a baculovirus vector: its use for the detection of bluetongue virus antibodies. AB - DNA representing RNA segment 3 of bluetongue virus (BTV) serotype 17, corresponding to the gene that codes for a group-specific antigen VP3, has been inserted into a baculovirus transfer vector in lieu of the 5' coding region of the polyhedrin gene of Autographa californica nuclear polyhedrosis virus (AcNPV). After cotransfection of Spodoptera frugiperda cells with wild-type AcNPV DNA in the presence of the derived recombinant transfer vector DNA, polyhedrin-negative recombinant baculoviruses were recovered. When S. frugiperda cells were infected with one of these recombinant viruses, a protein that was similar in size and antigenic properties to the BTV VP3 protein was synthesized. Antibodies raised in mice or rabbits to the baculovirus-expressed VP3 protein immunoprecipitated the VP3 protein of BTV-17 as well as that of BTV-10. The expressed antigen reacted with antisera representing four U.S.A. BTV serotypes in an indirect ELISA test. PMID- 3035064 TI - Surface structure and RNA-protein interactions of foot-and-mouth disease virus. AB - The surface structure of foot-and-mouth disease virus (FMDV) and the interaction of the individual capsid proteins with the virus RNA have been examined using modification reagents. By measuring the extent of modification of the lysine residues of intact and disrupted virus particles and the 12S protein subunit with Bolton & Hunter reagent it was found that 54% of the residues of VP1, 15% of the residues of VP2 and 37% of the residues of VP3, equivalent to five, two and four lysine residues respectively, are on the surface of the intact virus particle. Polypeptide VP4 was not modified in intact virus particles, indicating that it has no lysine residues on the surface of the virus. Modification with sodium metabisulphite, which causes a specific transamination reaction between cytidylic acid residues in ssRNA and closely associated basic amino acids, cross-linked all four structural proteins to the virus RNA. Both fragments of VP1, produced by treatment of the virus particle with trypsin, are also cross-linked to the RNA. These observations have been combined with the evidence that the immunogenic activity of VP1 may be contained in two discontinuous sites, at amino acids 141 to 160 and 200 to 213, in proposing a model for the arrangement of this polypeptide in the virus particle. PMID- 3035065 TI - Rat glial C6 cells are defective in murine coronavirus internalization. AB - Rat C6 glial cells were resistant to infection by several strains of murine coronaviruses. The restriction was not at the adsorption stage, since virus adsorbed to the C6 cells in a similar manner to mouse L cells which supported a lytic infection. The virus could not be internalized by the C6 cells. However, if the virus was introduced into the C6 cells by polyethylene glycol fusion, viral replication occurred and progeny virions were released from the infected cells. These studies indicated that the C6 cells were restrictive to coronavirus replication by preventing the early penetration stage of the viral replicative cycle. PMID- 3035066 TI - Sequence and N-terminal processing of the transmembrane protein E1 of the coronavirus transmissible gastroenteritis virus. AB - Sequencing of part of a clone from a transmissible gastroenteritis virus genome cDNA library led to the identification of the gene encoding the E1 matrix protein. The amino acid sequence of the primary translation product predicts a polypeptide of 262 residues which shares many features with the previously characterized murine hepatitis virus and infectious bronchitis virus E1 proteins. However, N-terminal amino acid sequencing revealed that a putative signal peptide of 17 residues was absent in the virion-associated polypeptide. The predicted mol. wt. of the mature unglycosylated product, 27,800, is in agreement with the experimental Mr value. PMID- 3035067 TI - Respiratory virus infection of peripheral blood monocytes: correlation with ageing of cells and interferon production in vitro. AB - The ability of respiratory syncytial virus (RSV) and parainfluenza virus type 3 (PIV3) to replicate in peripheral blood monocytes cultured in vitro for 1, 2, 4 or 7 days prior to infection was investigated. Inoculation of 1-day old monocytes produced at least tenfold less new virus than infection of the older, more macrophage-like cells for both viruses. PIV3 induced extensive syncytium formation, whereas RSV caused a cytopathic effect manifest by increased rounding of the cells with minimal syncytium formation. Supernatants of infected monocytes were assayed for human interferon-alpha (HuIFN-alpha) in an attempt to explain the restricted viral replication in the youngest monocytes. In PIV3-infected cells, HuIFN-alpha production was inversely correlated with new virus formation. Monocytes infected after 1 day in culture produced 800 IU/ml of HuIFN-alpha; the older cells produced 100 to 200 IU/ml. In contrast, monocytes infected on day 1 with RSV produced minimal amounts (1.5 IU/ml) of HuIFN-alpha. Increasing amounts of HuIFN-alpha were detected in cells infected with RSV after 2, 4 or 7 days in culture, reaching a maximum of 400 IU/ml on day 7. Further investigation of the apparent restriction of replication in young monocyte cultures may be helpful in understanding the pathogenesis of these respiratory infections. PMID- 3035068 TI - Mumps virus infection of dissociated rodent spinal ganglia in vitro. Expression and disappearance of viral structural proteins from neurons. AB - Cultured spinal ganglia and cord from mice and hamsters were infected with mumps virus or Sendai virus. Expression of five structural proteins, the haemagglutinin neuraminidase, fusion, nucleocapsid (NP), phospho (P) and matrix proteins was examined with monoclonal antibodies to each protein. In Sendai virus-infected mouse neurons all five viral proteins were detected. In hamster neurons infected with mumps virus all viral proteins were also expressed, but in mouse neurons only the NP and P proteins were seen. This suggests a species-specific cellular restriction of viral protein synthesis in mumps virus-infected mouse neurons. There was no, or only a slight, reduction in the number of neurons between days 4 and 20 after infection of mouse cultures with mumps virus, but the proportion of infected neurons diminished from 68% to 15% during this time. PMID- 3035069 TI - Latent herpes simplex virus type 1 DNA is not extensively methylated in vivo. AB - The methylation pattern of herpes simplex virus type 1 (HSV-1) DNA, present in the central nervous system of latently infected mice, was examined by digestion of the DNA with methylation-sensitive restriction endonucleases and Southern blot hybridization. Using the enzymes SmaI, XmaI, SalI and SacII, the data indicate no extensive methylation of latent HSV-1 DNA in vivo. Thus, extensive methylation of the viral genome is not a necessary condition for, or a consequence of maintaining, the latent state in vivo. PMID- 3035070 TI - Diagnosis of enterovirus 70 infection by demonstration of IgM antibodies. AB - Since outbreaks of severe acute hemorrhagic conjunctivitis occur worldwide [Hierholzer and Hatch, 1985] and the majority of the epidemics are caused by enterovirus 70 (EV-70), we developed an EV-70 IgM ELISA to simplify the diagnosis of these outbreaks. The test is based on the capture antibody technique and the use of monoclonal antibodies to EV-70. We detected EV-70 IgM antibodies in 55% of 76 convalescent-phase sera from an outbreak of acute hemorrhagic conjunctivitis in a Brazilian community. Among the 71 acute- and convalescent-phase serum pairs from this outbreak, 49 (69%) demonstrated a 4-fold or greater rise in neutralizing antibody-titer. The titer of IgM antibody began to drop by the fifth week after onset of illness. EV-70 IgM antibodies were not detected in 53 serum pairs with a 4-fold or greater rise in antibodies to other picornaviruses. The EV 70 ELISA proved to simple and relatively rapid to perform, appeared to be specific, and should be sensitive enough to diagnose outbreaks of EV-70 when multiple serum specimens can be tested. PMID- 3035071 TI - Use of monoclonal antibodies for the diagnosis of cytomegalovirus infection by the detection of early antigen fluorescent foci (DEAFF) in cell culture. AB - A pool of seven monoclonal antibodies, each reactive with cytomegalovirus (CMV) early antigens, was used in an indirect immunofluorescence method for the rapid detection of CMV-infected fibroblasts following inoculation with clinical specimens. A total of 1,639 specimens were examined, and the results were compared with those of conventional isolation procedures. The detection of CMV by early antigen fluorescent foci (DEAFF) was found to be comparable, both in terms of specificity and sensitivity, to that of conventional cell culture. Its great advantage, however, is the rapidity with which results are achieved. Thus, results were available from the DEAFF test within 24 hours of receipt of the specimens as compared to a mean of 16 days for cell culture. This single rapid assay for the detection of CMV in clinical samples may be performed by any laboratory familiar with cell culture techniques and in our hands is the preferred diagnostic method for CMV. PMID- 3035072 TI - Analysis by immunoblotting of human cytomegalovirus antibody in sera of renal transplant recipient. AB - Human cytomegalovirus-infected cell polypeptides were immunoreacted by sera of renal transplant recipients and compared with those reactive with sera of healthy adult donors by means of the Western immunoblotting technique. At least 15 polypeptides with molecular weights of 155K, 123K, 102K, 89K, 79K, 71K, 65K, 60K, 55K, 50K, 46K, 42K, 38K, 33K, and 28K were immunoreacted. Sera obtained serially from renal transplant recipients reacted with most of these polypeptides and reacted more frequently and intensely with the smaller polypeptide species such as 38K, 33K, and 28K, compared with sera of healthy seropositive adults. The implications of these findings are discussed. PMID- 3035073 TI - Seroepidemiological survey of the prevalence of antibodies to a strain of human calicivirus. AB - Batches of pooled immune globulins and sera were tested by immune electron microscopy (IEM) for the presence of antibodies to a strain of human calicivirus (HCV, UK1). The results show that this strain of HCV is prevalent throughout many parts of the world and that the majority of the population experience infection by the age of 12 years. The survey carried out in the United Kingdom indicates that the presence of maternal antibody correlates with some degree of protection during the first few weeks of life; the peak incidence of cases and acquisition of antibody occur between 3 months and 6 years. Tests on sera from Japan show a similar pattern of acquisition of antibodies and demonstrate that infection with more than one strain of HCV commonly occurs during childhood. PMID- 3035076 TI - Serum and urine concentrations of oral bromovinyldeoxyuridine in humans as monitored by a bioassay system based on varicella-zoster virus focus inhibition. AB - A simple and sensitive bioassay method for measuring (E)-5-(2-bromovinyl)-2' deoxyuridine (BVDU) concentrations in human serum and urine has been established. This method is based on the inhibitory effect of BVDU on varicella-zoster virus (VZV) focus formation in vitro. The minimal concentration of BVDU that could be detected in serum by this method was 0.2 microgram/ml. Following a single oral administration of 250 mg BVDU, serum BVDU concentrations of 1.2-2.2 micrograms/ml were attained 1 hr later; at 5 and 7 hr, serum BVDU levels were below 0.2 microgram/ml. Upon repeated administration of 125 mg BVDU at 8 hr intervals, the serum BVDU concentrations reached 0.7-1.1 microgram/ml at 2 hr after the fourth administration. These concentrations are approximately 300-450-fold higher than the 50% inhibitory dose of BVDU for VZV in vitro. Urinary BVDU concentrations were on average 10 to 20 times higher than the serum BVDU concentrations. PMID- 3035075 TI - Detection of specific types of human papillomavirus in cervical scrapes, anal scrapes, and anogenital biopsies by DNA hybridization. AB - Specific varieties of human papillomavirus (HPV) infecting the anogenital region were detected in clinical samples by use of a filter hybridization technique suitable for rapid screening of cervical and anal scrapes. In this way possibly benign types (HPV6 and HPV11) could be differentiated from types thought to be capable of malignant transformation (HPV 16 and HPV 18). Cervical or anal canal cells were applied directly to nylon filters and fixed by u.v. irradiation before hybridization with mixed viral DNA probes under both low- and high-stringency conditions. In addition, probe for the human Alu-repeated DNA sequence was used to assess the relative amount of total nucleic acids in each sample applied to the filter. HPV DNA was detected in 3 of 19 cervical scrapes from patients with no past or present history of wart virus infection or cervical dysplasia. Within a positive study group totalling 71 patients, HPV (6/11 or 16/18) was detected in cervical scrapes from 24% of 41 patients who did not have visible genital dysplasia, 30% of 27 patients with visible genital dysplasia or cervical intraepithelial neoplasia (CIN) I, and in 1 of 3 patients with past CIN II/III. In addition, HPV6/11 or 16/18 DNA was detected in anal scrapes from 3 of 6 male patients and in 85% of genital biopsies. A notably high proportion (4/6) of vaginal condylomata were positive with both the HPV6/11 and the HPV16/18 mixed viral DNA probes. Of the biopsies prepared for histopathology and positive for HPV DNA, the HPV group-specific antigen could be detected in only 60%. PMID- 3035074 TI - Serological study of rubella-like illnesses. AB - We investigated 627 patients who within a period of 2 1/2 years had had a rubelliform rash and/or symptoms of arthritis and arthralgia. Sera from these patients were investigated for evidence of rubella, human parvovirus B19 (HPV), and measles infection with methods to detect specific IgM and IgG antibodies. Complement fixation tests were used to screen for a wide range of other infectious agents. We detected 229 cases of rubella, 43 cases of HPV infection, 7 cases of measles, and 9 cases of infection by various other aetiological agents. This left a large proportion of rubelliform rashes, 54% (339 cases), whose aetiology was unknown. This study confirmed that the diagnosis of rubella on clinical grounds alone is unreliable. Many (6.8%) of the rashes in the study were due to HPV infection, and the seasonal incidence was the same as for rubella. There was considerable overlap between the features of rubella and HPV infections, although in adults arthralgia occurred more frequently in HPV infections than in rubella. In all cases HPV infection was self-limiting, although, as in rubella, symptoms can be prolonged and one adult's disease lasted almost 9 months. Purpura was noted in only one patient with HPV infection. In this study three patients had HPV infection during pregnancy. Two patients spontaneously aborted one month later. The third patient progressed to full term and delivered a healthy baby. PMID- 3035077 TI - Spontaneous and induced interferon production by peripheral blood leucocytes from control population and patients with herpes genitalis. AB - The spontaneous and PHA induced levels of interferon were measured in peripheral blood leucocyte cultures of twenty-eight individuals diagnosed as positive for herpes genitalis, and in a group of control subjects. As reported by Cunningham and Merigan [1983] for herpes labialis, leucocytes from individuals with herpes genitalis produced low levels of interferon spontaneously; however, similar results were found for individuals within the control population. No statistically significant difference could be found for PHA induced interferon levels, antigen induced interferon levels, or helper/suppressor cell ratios between the herpes genitalis population and control population. Our results indicate that interferon does not play a major role in the latency or recurrence of herpes genitalis. PMID- 3035078 TI - Cytopathology, plaque assay, and heat inactivation of hepatitis A virus strain HM175. AB - Hepatitis A virus (HAV) strain HM175 derived after repeated subculture of persistently infected B-SC-1 cells caused a specific cytopathic effect upon acute infection of B-SC-1 cells. The virus formed visible plaques on B-SC-1 cell monolayers after 9 to 14 days of incubation at 34 degrees C, and virus can therefore be titrated by plaque assay. Virus could be re-isolated from plaques of infected cells, which allows the clonal isolation of HAV variants. The stability of a plaque-purified variant of HAV at elevated temperatures exceeded that of poliovirus type 1, but this variant is less stable than previously reported strains of HAV. PMID- 3035079 TI - Development of a plaque assay for a cytopathic, rapidly replicating isolate of hepatitis A virus. AB - Most hepatitis A virus (HAV) replication in cell culture has been reported to be nonlytic and relatively slow. A rapidly replicating isolate of strain HM-175 from persistently infected, serially passed cell cultures (pHM-175) was found to induce a cytopathic effect. This observation allowed the development of a classic plaque assay for pHM-175 in FRhK-4 cells. The plaques were neutralized by polyclonal and monoclonal antisera to HAV. PMID- 3035080 TI - Effect of antiviral substances on hepatitis A virus replication in vitro. AB - The effect of protamine, atropine, selenocystamine, taxifolin, and catechin on the infectivity and antigenicity of the cell culture-adapted hepatitis A virus (HAV) strain CF 53 was studied. The toxicity on uninfected PLC/PRF/5 cells was examined for each antiviral compound by morphological and biochemical methods, in order to determine concentrations without cytotoxic effect. At these concentrations, protamine and taxifolin, added to infected cells for a 15-day period, caused concentration-dependent reductions in the infectivity and antigenicity of HAV. Atropine also caused a concentration-dependent reduction of HAV infectivity but did not affect the antigenicity of the virus. At the highest concentration used, 50 micrograms/ml of protamine, 59 micrograms/ml of taxifolin, and 50 micrograms/ml of atropine, the infectious viral titer reduction was 1.56, 0.77, and 0.68 log10, respectively. Selenocystamine and catechin had no effect on HAV replication. PMID- 3035081 TI - An enzyme labelled nuclear antigen immunoassay for detection of cytomegalovirus IgM antibodies in human serum: specific and non-specific reactions. AB - A mu-capture enzyme linked immunosorbent assay was developed for detection of IgM antibody to cytomegalovirus (CMV). Virus-specific IgM was detected using horseradish peroxidase labelled nuclear CMV antigen (CMV-ELA). False-positive reactions caused by Paul-Bunnell-Davidsohn (PBD) positive sera and antinuclear antibody (ANA) positive sera were identified in a combination assay employing enzyme labelled nuclear control antigen (CO-ELA) in parallel to the CMV-ELA. Four of five PBD positive and 30 of 31 ANA positive sera reactive with the CMV-ELA were identified as false positive reactions in the combined ELA-assay. The reactivity in PBD-positive sera could not be explained by antigenic cross reactivity between CMV and Epstein-Barr virus, and the results further suggested that different cell specified components of the CMV-ELA were responsible for the reactivity of PBD-positive as compared to ANA-positive sera. One of 314 healthy blood donors, 12 of 12 patients with primary CMV infection, and 11 of 15 patients with secondary CMV infection had detectable CMV IgM antibodies. Comparison of different CMV-ELAs revealed that pronounced differences in specificity as well as sensitivity may exist. PMID- 3035082 TI - Immunogenicity of a hepatitis A virus vaccine. AB - Hepatitis A virus (HAV) strain HAV/HFS/GBM was adapted to and grown in human diploid fibroblast cells. The HAV was concentrated by ammonium sulphate precipitation and high-speed centrifugation. The virus was inactivated by beta propiolacton and purified by sedimentation through a 20% solution of sucrose and by CsCl gradient centrifugation. The immunogenicity of different preparations was tested in mice, guinea pigs, and goats. The immune response after vaccination was tested by determination of anti-HAV with RIA and of neutralizing antibodies with an appropriate test system. Results showed that anti-HAV titers of 1:1,000 and greater, as well as neutralizing antibody titers of 1:1,000 and greater, were found in sera of animals vaccinated with different preparations. It became evident that good anti-HAV titers persisted over a period of at least 1.5 years in goats and mice after immunization with a semipurified HAV vaccine, and titers up to 1:200 were present in mice 2 years after vaccination with a highly purified HAV vaccine. PMID- 3035083 TI - Serotonergic influence on the growth hormone response to clonidine in rat. AB - Administration of the alpha 2-adrenoceptor agonist clonidine induces growth hormone (GH) release in rat and man. In the present study it is shown that the GH response to clonidine is weaker in rats exposed to depletion of both noradrenaline and serotonin (by means of reserpine or the combined treatment of FLA-63 and PCPA) than in animals exposed to noradrenaline depletion (by means of FLA-63) only. The possibility that an impaired serotonergic neurotransmission contributes to the blunted GH responses to clonidine observed in patients suffering from endogenous depression is discussed. PMID- 3035084 TI - Effects of GABAergic drugs on the GABA turnover in the substantia nigra and the corpus striatum of the rat. AB - Changes in the endogenous GABA concentration and in GABA turnover following GABA receptor stimulation or blockade were studied in the substantia nigra and the corpus striatum of the rat. The GABA agonists, muscimol, baclofen and THIP decreased the accumulation of GABA following inhibition of GABA-alpha ketoglutaric acid aminotransferase by gamma-acetylenic GABA (GAG) in both structures investigated. Only the effect of muscimol in the substantia nigra was inhibited by the GABA antagonist, bicuculline. Muscimol, baclofen and progabide reduced the disappearance rate of GABA in the substantia nigra following inhibition of the glutamate decarboxylase by 4-deoxypyridoxine. The endogenous GABA concentration was decreased in the corpus striatum following muscimol, THIP or baclofen, probably due to a decreased synthesis of GABA. Smaller effects were seen on the endogenous GABA concentration in the substantia nigra, since both the synthesis and the utilization of GABA were decreased by muscimol and baclofen. Thus, the turnover of brain GABA might be regulated by changes in receptor activity. PMID- 3035086 TI - [3H] UK-14,304, a new agonist ligand of alpha 2-adrenoceptors: a comparative study with human and rat tissue. AB - [3H] UK-14,304 was used to investigate alpha 2-adrenoceptors in rat brain and human platelets. Receptor pharmacology revealed that the ligand binds with high affinity to alpha 2-adrenoceptors. Psychoactive substances like neuroleptics, antidepressants, and beta-carbolines displace [3H] UK-14,304 from its binding sites in the lower micromolar range. A Hill number around 2 for most neuroleptics suggests a positive cooperativity with the alpha 2-adrenoceptors. Comparative studies with [3H] UK-14,304 and [3H] clonidine utilizing platelet membranes from human volunteers demonstrated that the former ligand is more suitable to investigate possible changes of alpha 2-adrenoceptors; [3H] UK-14,304 labels more receptors with a lower standard deviation, whereby the volume of the blood sample amounted to 35 ml instead of 50 ml required for [3H] clonidine as ligand. No sex differences of binding constants were detected, however an inverse correlation of maximum number of binding sites and affinity was found for female subjects with both ligands. No age-dependent changes of Bmax and KD-values were observed in the range of 24 to 59 years. PMID- 3035085 TI - Locus coeruleus neurons show reduced alpha 2-receptor responsiveness and decreased basal activity in spontaneously hypertensive rats. AB - Previous studies have indicated that brain noradrenaline (NA) neurons in spontaneously (genetically) hypertensive rats (SHR) are implicated in the development of hypertension. Thus, a number of biochemical aberrations in the metabolism of NA in the SHR brain have been detected although the data are not in total agreement. We report here experiments utilizing single cell recording techniques which show directly a reduction in neuronal activity of brain NA neurons in the locus coeruleus (LC) of SHR. This reduction develops gradually with age and in parellel with the increased blood pressure (BP), but is not altered by acute alterations in BP. The SHR were found to display an increased intraneuronal monoamine oxidase (MAO) activity as well as a specifically reduced sensitivity of inhibitory alpha 2-receptors within the LC. It is suggested that in SHR the LC system, in spite of a reduced basal activity displays increased responsiveness to sensory stimuli, a phenomenon that may contribute to the development of hypertension. PMID- 3035088 TI - Kinetics of pyrophosphate induced iron release from diferric ovotransferrin. AB - The kinetics of pyrophosphate-induced iron release from diferric ovotransferrin were studied spectrophotometrically at 37 degrees C in 0.1 M HEPES, pH 7.0. At high pyrophosphate concentrations, the kinetics are biphasic, indicating that the rates of iron release from the two, presumably noninteracting iron-binding sites of ovotransferrin are different. The pseudo-first-order rate constants for iron release from both the fast and slow sites exhibit a hyperbolic dependence on pyrophosphate concentrations. The data suggest that pyrophosphate forms complexes with the two iron-binding sites of ovotransferrin prior to iron removal. The stability constants of the complex formed with the fast site (Keqf) and slow site (Keqs) are 8.3 M-1 and 40.4 M-1, respectively. The first-order rate constants for the dissociation of ferric-pyrophosphate from the fast site (k2f) and the slow site (k2s) are 0.062 and 0.0044 min-1, respectively. Results from urea gel electrophoresis studies suggest that iron is released at a much faster rate from the N-terminal binding site of ovotransferrin. At high pyrophosphate concentration, only C-monoferric-ovotransferrin is detected during the course of iron release. At low pyrophosphate concentration, however, a detectable amount of N-monoferric-ovotransferrin is accumulated. This result is consistent with the kinetic finding that the site with a higher k2 (0.062 min-1) has a lower affinity toward pyrophosphate (Keq = 8.3 M-1) whereas the site with a lower k2 (0.0044 min 1) has a higher affinity for pyrophosphate (Keq = 40.4 M-1). PMID- 3035089 TI - Cerebral phosphoinositide and energy metabolism during and after insulin-induced hypoglycemia. AB - During and after insulin-induced hypoglycemia, changes in levels of cerebral phosphatidylinositol (PI), phosphatidylinositol 4-phosphate (PIP), phosphatidylinositol 4,5-bisphosphate (PIP2), phosphatidic acid (PA), triacylglycerol (TAG), diacylglycerol (DAG), and free fatty acids (FFAs) as well as the cerebral energy state were studied in relation to the EEG. In hypoglycemic rats with an EEG pattern of quasiperiodic sharp or slow sharp waves, which preceded the development of an isoelectric EEG, PIP2 levels increased significantly, together with a slight decrease in PI content. Levels of the other lipids did not change during this period. The cerebral energy state was affected only slightly in spite of profound decreases in plasma and tissue glucose levels. With 30 min of an isoelectric EEG, levels of all phosphoinositides and PA decreased significantly; total FFA and DAG contents increased seven- and twofold, respectively; the TAG-palmitate level decreased, and that of TAG-arachidonate increased. Plasma and tissue glucose were nearly depleted, and the cerebral energy state deteriorated severely. The increment in fatty acids in the DAG and FFA pools was less than their loss from phosphoinositides and PA, an observation suggesting vascular washout or oxidation of a portion of the FFAs produced. Following 90 min of glucose infusion, PIP and PA levels recovered to control values; however, the PIP2 content exceeded control levels, and that of PI remained below control levels. DAG and FFA contents returned to normal.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3035087 TI - Disturbed function of the pineal gland in familial amyloid polyneuropathy. AB - In order to estimate the function of the pineal gland, the sympathetic nervous system and the adrenal medulla in patients with familial amyloid polyneuropathy relative to healthy subjects, we have quantified urinary 6-hydroxymelatonin, normetanephrine and metanephrine. Urinary 6-hydroxymelatonin level correlated with neither of two O-methylated catecholamine levels in healthy subjects. The excretion of both 6-hydroxymelatonin and metanephrine were reduced in the patient group as compared with the control group, and the normal daily rhythm of 6 hydroxymelatonin was undetectable in the most of the patients. This finding indicates that the function of the pineal gland is disturbed in familial amyloid polyneuropathy. PMID- 3035090 TI - Effects of an extract of Ginkgo biloba on the 3',5'-cyclic AMP phosphodiesterase activity of the brain of normal and triethyltin-intoxicated rats. AB - For clarification of the beneficial effects of the extract of Ginkgo biloba (EGB) on triethyltin (TET) toxicity in rats, the phosphodiesterase (PDE) activities of the cerebral tissue were measured under in vitro and ex vivo conditions. Under in vitro conditions, low concentrations of EGB (0.25-4.0 mg/L) activated the enzyme, whereas after higher concentrations (5-250 mg/L), dose-dependent inhibition of the enzyme activity was observed. In the lower concentration range, the extract also partially restored the high-affinity PDE activity (measured with 0.25 microM cyclic AMP) of the particulate fraction of the brain inhibited by TET in vitro. In contrast, the inhibitory influence of TET on the low-affinity PDE activity (measured with 50 microM cyclic AMP) of the particulate fraction was enhanced by the extract. Although treatment with a single large dose of EGB lowered the particulate PDE activities of the brain of normal rats, no effects of the extract could be detected in animals after repeated daily administrations of EGB during a 4-day period. Curative treatment of the TET-intoxicated rats with EGB during a 7 day period accelerated the recovery of the edematous state of the white matter caused by the intoxication and also normalized the lowered PDE activity of the particulate fraction of the edematous brain tissue. Furthermore, when preventively administered, EGB counteracted both the edema formation and the fall in PDE activity observed with treatment by TET alone. These observations strongly suggest that some beneficial effects of EGB might be due to its modulating influences on cellular cyclic AMP levels via activation of membrane-bound PDE. PMID- 3035092 TI - Effects of antidepressant drug treatments on alpha 2-adrenoceptor control of [3H]noradrenaline release from hypothalamic synaptosomes. AB - KCl (16 mM) stimulated the release of [3H]noradrenaline ([3H]NA) from rat hypothalamic synaptosomes in a Ca2+-dependent manner; this release was attenuated by clonidine (0.01-100 microM). Changes in the release of [3H]NA and the functional status of alpha 2-adrenoceptors in the medial hypothalamus of rats treated acutely and chronically with clorgyline (1 mg/kg/day) or desipramine (DMI, 10 mg/kg/day) were assessed using superfused synaptosomes in which the attenuating effects of clonidine (1 microM) or the potentiating effects of yohimbine (1 microM) on K+-evoked release of [3H]NA were measured. After acute administration of DMI, significantly less [3H]NA was accumulated into synaptosomes. Although total (spontaneous + K+-evoked) [3H]NA release from these synaptosomes was unchanged, a significant reduction was apparent in the K+-evoked release from the DMI-treated tissue. Attenuation of K+-evoked release by clonidine was abolished in both these acute treatment groups. Following the chronic antidepressant drug regimens, [3H]NA uptake into DMI-treated tissue remained significantly reduced although total percent and K+-evoked [3H]NA release were unchanged. The K+-evoked release of [3H]NA in S1 was significantly enhanced (by 22%) in the clorgyline treatment group. Attenuation of K+-evoked [3H]NA release by clonidine in both chronic antidepressant-treated tissues was not significantly changed. It is concluded that the functional sensitivity of alpha 2-adrenoceptors on nerve endings in the medial hypothalamus is unchanged by these chronic antidepressant drug regimens. In synaptosomes from untreated tissue, yohimbine significantly potentiated K+-evoked release of [3H]NA; this effect was unchanged after acute regimens and reduced after chronic administration of both the antidepressants.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3035093 TI - Activation of dopamine receptors does not affect phosphoinositide turnover in NCB 20 cells. AB - Dopamine inhibits and serotonin stimulates adenylate cyclase activity in a neuroblastoma X Chinese hamster brain explant cell line (NCB-20). The inhibition of cyclic AMP accumulation by dopamine was blocked by pretreatment of the cells with pertussis toxin. Carbachol and bradykinin stimulated the accumulation of water-soluble inositol phosphates whereas thyrotropin-releasing hormone, vasopressin, neurotensin, and phenylephrine were without effect. Dopamine and serotonin had no significant effect on carbachol-induced phosphoinositide hydrolysis or the levels of the parent lipids within the membrane. Forskolin induced a much larger stimulation of cyclic AMP than did serotonin, and caused an increase in the levels of phosphatidylinositol-4-phosphate and phosphatidyl inositol-4,5-bisphosphate in the cell membrane. PMID- 3035091 TI - Endogenous dopamine functionally activates D-1 and D-2 receptors in striatum. AB - Rat striatal slices incubated with the phosphodiesterase inhibitor 3-isobutyl-1 methylxanthine at 1 mM were exposed to different concentrations (1-100 microM) of the catecholamine-releasing drug amphetamine. This produced both a concentration dependent release of endogenous dopamine and accumulation of cyclic AMP in the slices. The cyclic AMP accumulation due to amphetamine was greatly increased when slices were coincubated with the selective dopamine D-2 antagonist (-)-sulpiride (30 microM), but the amphetamine-induced release of dopamine from the slices was the same in the presence or absence of (-)-sulpiride. Pretreatment of animals with reserpine (5 mg/kg s.c., 18 h before death) and in vitro incubation with alpha-methyl-p-tyrosine (50 microM for 90 min), respectively, reduced the ability of amphetamine (1-100 microM) [in the presence of 30 microM (-)-sulpiride] to induce release of dopamine and to elevate cyclic AMP accumulation in striatal slices. A similar reduction in amphetamine-induced dopamine release and cyclic AMP accumulation in striatal slices was observed 7 days following unilateral 6 OHDA lesions of the medial forebrain bundle of rats. These results suggest that amphetamine induces release of endogenous dopamine from the terminals of nigrostriatal dopamine neurones. Released dopamine is then able functionally and concomitantly to activate D-1 and D-2 receptors, seen as stimulation and inhibition of cyclic AMP accumulation, respectively. PMID- 3035094 TI - Ionic mechanisms implicated in the stimulation of cerebellar cyclic GMP levels by N-methyl-D-aspartate. AB - N-Methyl-D-aspartate (NMDA) increases cyclic GMP levels in immature rat cerebellar slices incubated in magnesium-containing Krebs buffer in vitro. This effect is blocked by 2-amino-5-phosphonovalerate and by D-alpha-aminoadipate, but not by glutamic acid diethyl ester or gamma-D-glutamylaminomethylsulfonic acid, indicating specific involvement of the NMDA receptor. The response produced by NMDA is abolished by removal of calcium from the medium, proportional to the concentration of extracellular calcium, and blocked by a number of inorganic (Ni2+, Co2+, Cd2+, La3+, Mn2+) calcium antagonists. The responses to NMDA are not blocked by barium or strontium and persist when these ions are substituted for calcium in the incubation medium. The effects of NMDA are blocked by, but are not particularly sensitive to, the organic voltage-dependent calcium channel antagonists. Nifedipine (10 microM) produces partial inhibition of the effects of NMDA, which are also antagonized by high (greater than 200 microM) concentrations of diltiazem and verapamil. The effects of NMDA are tetrodotoxin insensitive but are abolished by omission of sodium from the medium and inhibited by a tetrodotoxin-insensitive sodium channel blocker, Zn2+. The results suggest that calcium channel opening is a consequence of NMDA receptor activation in this model. However, the sodium dependence of the response argues against the use of receptor-operated calcium channels, whereas the weak activity of the organic voltage-sensitive calcium channel antagonists argues either against the use of voltage-dependent calcium channels, or that those implicated in the effects of NMDA are insensitive to these agents.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3035095 TI - Effect of exogenous gangliosides on amino acid uptake and Na+, K+-ATPase activity in superior cervical and nodose ganglia of rats. AB - The effects of some gangliosides on active uptake of nonmetabolizable alpha aminoisobutyric acid (AIB) and Na+, K+-ATPase and Ca2+, Mg2+-ATPase activities in superior cervical ganglia (SCG) and nodose ganglia (NG) excised from adult rats were examined during aerobic incubation at 37 degrees C for 2 h. In NG, amino acid uptake was greatly accelerated with the addition of galactosyl-N acetylgalactosaminyl-[N-acetylneuraminyl]-galactosylgluc osyl ceramide (GM1) (85%) and also with N-acetylgalactosaminyl-[N-acetylneuraminyl] galactosylglucosyl ceramide (GM2) or [N-acetylneuraminyl]-galactosyl-N acetylgalactosaminyl-[N-acetyl- neuraminyl]-galactosylglucosyl ceramide (GD1a) (43% each) compared with a nonaddition control at a 5 nM concentration. Under identical conditions, Na+, K+-ATPase activity was strongly stimulated with GM1 (180%) and GD1a (93%), whereas Ca2+, Mg2+-ATPase activity showed no change. In SCG, on the other hand, AIB uptake was apparently inhibited (-27%) by addition of GM1, with a slight decrease in Na+, K+-ATPase but no change in Ca2+, Mg2+-ATPase activity in the tissue. Both asialo-GM1, in which N-acetylneuraminic acid is deficient, and Forssman glycolipid, which is not present in nervous tissue, failed to produce any significant increase in both SCG and NG not only in amino acid uptake, but also in Na+, K+-ATPase activity. A kinetic study of active AIB uptake showed that GM1 ganglioside produced an increase in Km with no change in Vmax in SCG, whereas it caused a decrease in Km with a slight increase in Vmax in NG. Treatment of NG and SCG with neuraminidase from Vibrio cholerae, an enzyme that split off sialic acid from polysialoganglioside, leaving GM1 intact, caused little inhibition of the amino acid uptake.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3035096 TI - Secretion of [Met]enkephalyl-Arg6-Phe7-related peptides and catecholamines from bovine adrenal chromaffin cells: modification by changes in cyclic AMP and by treatment with reserpine. AB - Investigations into the effects of culturing bovine adrenal chromaffin cells in the presence (72 h) of dibutyryl cyclic AMP, forskolin, and reserpine on the level and release of [Met]enkephalyl-Arg6-Phe7 immunoreactivity, noradrenaline, and adrenaline are reported. The assay for [Met]enkephalyl-Arg6-Phe7 immunoreactivity recognises both peptide B, the 31-amino acid carboxy-terminal segment of proenkephalin, and its heptapeptide fragment, [Met]enkephalyl-Arg6 Phe7. Treatments that elevate cyclic AMP increase the amount of peptide immunoreactivity in these cells; this is predominantly peptide B-like immunoreactivity in both control cells and cyclic AMP-elevated cells. Treatment with reserpine gives no change in total immunoreactivity levels, but does not result in increased accumulation of the heptapeptide [Met]enkephalyl-Arg6-Phe7 at the expense of immunoreactivity that elutes with its immediate precursor, peptide B. Cyclic AMP treatment causes either no change or a decrease in levels of accumulated noradrenaline and adrenaline. However, the release of [Met]enkephalin Arg6-Phe7 immunoreactivity, noradrenaline, and adrenaline is increased by 72-h pretreatment with forskolin or dibutyryl cyclic AMP, whether release is stimulated by nicotine or elevated potassium. In each case the molecular form of [Met]enkephalyl-Arg6-Phe7 immunoreactivity that is released approximately reflects the cell content. Pretreatment with reserpine has no effect on the total [Met]enkephalyl-Arg6-Phe7 immunoreactivity released, but does result in an increased release of the heptapeptide and a decrease in release of peptide B-like immunoreactivity. The studies suggest that the levels of [Met]enkephalyl-Arg6 Phe7 and peptide B available for release are controlled both at the level of proenkephalin synthesis and at the level of double-basic residue proteolysis. PMID- 3035097 TI - Lack of usefulness of DN-1417 for characterization of a CNS receptor for thyrotropin-releasing hormone. AB - CNS receptors for thyrotropin-releasing hormone (TRH) and its analogs are likely to mediate the experimentally and clinically observed net excitatory effect of these peptides on lower motor neurons. Previous findings suggest that several types of TRH receptors with distinct TRH analog specificities may be present in rat CNS. In particular, based on competition isotherm assays with unlabeled analog gamma-butyrolactone-gamma-carbonyl-L-histidyl-L-prolineamide (DN-1417). Funatsu et al. claim the existence of a limbic forebrain site that binds this peptide and TRH with high affinity but that does not bind [3-methyl-histidyl2] TRH (MeTRH). Using saturation and competition isotherm experiments, we have examined the binding of [3H]TRH and [3H]DN-1417 in three regions of rat CNS: pyriform cortex/amygdala, limbic forebrain, and lumbosacral spinal cord. In all three regions, saturation assays with [3H]TRH (0.4-100 nM) resolved only a single, saturable receptor with high affinity (KD = 12-14 nM) for TRH; in no case could more than one saturable site be identified. When [3H]DN-1417 was substituted as the assay ligand, no high-affinity binding component for this analog could be detected in the three regions. Competition curves for the binding of unlabeled DN-1417 to limbic forebrain and lumbosacral spinal cord ([3H]TRH as assay ligand) were monophasic (not biphasic like those of Funatsu et al.) and indicative of low-affinity binding of DN-1417 in these regions (Ki values = 2-3 microM; in agreement with values obtained in similar assays with [3H]MeTRH).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3035098 TI - Effect of the tricyclic antidepressant desipramine on beta-adrenergic receptors in cultured rat glioma C6 cells. AB - Rat glioma C6 cells, cultured in the presence of the tricyclic antidepressant desipramine, lost a significant number of beta-adrenergic receptors in a time- and dose-dependent manner. A similar loss was observed whether binding was determined on intact cells with the hydrophilic beta-adrenergic antagonist (+/-) [3H]4-(3-tert-butylamino-2-hydroxypropoxyl)benzimidazole-2-o n HCl ([3H]CGP 12177) or on cell lysates with the more hydrophobic antagonists [125I]iodocyanopindolol or [3H]dihydroalprenolol. When stimulated with the agonist isoproterenol, desipramine-treated cells accumulated less cyclic AMP than control cells. The affinity of the beta-adrenergic receptors for either antagonist or agonist was unchanged after desipramine treatment. Desipramine interacted only weakly with the receptors and competed for [125I]iodocyanopindolol binding with a Ki of 30 microM. The presence in the culture medium of alprenolol or propranolol, potent beta-adrenergic antagonists, however, did not prevent the reduction in receptors by desipramine. Desipramine also caused a loss of beta-adrenergic receptors from cells maintained in serum free medium and the cells themselves did not contain or secrete endogenous catecholamines. Although desipramine is a potent inhibitor of catecholamine uptake, it appears unlikely that the observed loss of beta-adrenergic receptors in rat glioma C6 cells exposed to the drug is due to an increase in extracellular catecholamine levels or to a direct interaction with the receptors. PMID- 3035099 TI - Depletion of myo-inositol and amino acids in galactosemic neuropathy. AB - A depletion of not only myo-inositol (MI) but also taurine and other amino acids was observed in the sciatic nerve of a galactosemic rat. Treatment of the galactosemic rats with sorbinil, an aldose reductase (AR) inhibitor, was found to block galactitol formation and protect against the loss of MI, taurine, and other amino acids. Incubation studies of sciatic nerve have revealed that [3H]MI and [3H]taurine were actively taken up and concentrated. Incubation of the nerve in a high-galactose medium showed a decrease in the accumulation of [3H]MI and [3H]taurine whereas the galactitol level increased. Time-course studies have shown that the galactitol level reached a plateau before a substantial decrease in the accumulation of [3H]MI and [3H]taurine occurred. The addition of AR inhibitors in the galactose medium significantly protected against the loss in the capacity of the nerve to accumulate [3H]MI and [3H]taurine. Hypertonicity of the galactose medium also seemed to have a protective effect similar to that of AR inhibitors. PMID- 3035100 TI - Effects of systemic kainic acid administration on regional Na+, K+-ATPase activity in rat brain. AB - Changes in the activity of Na+,K+-ATPase and in the water, Na+, and K+ levels in the parietal cortex, hippocampus, and thalamus were investigated in rats 1, 3, 6, and 24 h following systemic kainic acid injection. An increase in Na+,K+-ATPase activity was observed in all three regions 3 h after the treatment, with a subsequent decrease in enzyme activity. The elevation in Na+,K+-ATPase activity was accompanied by an increase in the Na+ content and a decrease in the K+ content. These changes are presumed to occur because of repeated discharges and excessive prolonged depolarization in response to kainic acid. The decreases in Na+,K+-ATPase activity 6 and 24 h following kainic acid treatment coincide with neuropathological damage and edema formation, mainly in the hippocampus and thalamus. PMID- 3035101 TI - Thyrotropin-releasing hormone reduces myo-inositol content in rat cerebellum pretreated with lithium. AB - The effect of thyrotropin-releasing hormone (TRH) and lithium on myo-inositol metabolism has been assessed in rat cerebral cortex, cerebellar cortex, and sciatic nerves. Sprague-Dawley male rats were injected subcutaneously with 10 mEq/kg of LiCl and intraperitoneally with 10 mg/kg of TRH-tartrate, alone or in combination. Either lithium or TRH alone had little effect on the myo-inositol concentration in cerebellar cortex, whereas the combination of lithium and TRH significantly lowered the level. The myo-inositol level of cerebellar cortex reached its nadir (70% of values in untreated control rats) 30 min after addition of TRH and then returned to the control level at 90 min. In cerebral cortex, both lithium alone and lithium plus TRH significantly reduced the myo-inositol level. No effect was seen on the myo-inositol concentration in sciatic nerves with these regimens. These results suggested that the pharmacological dose of TRH activated phosphatidylinositol turnover in rat cerebellar cortex and subsequently reduced the myo-inositol level in the presence of lithium. PMID- 3035102 TI - The flexor carpi radialis H-reflex in polyneuropathy: relations to conduction velocities of the median nerve and the soleus H-reflex latency. AB - In 80 controls latencies of flexor carpi radialis (FCR) and in 94 controls latencies of soleus H-reflexes correlated well with length of the extremity, body height and age. Multiple regression equations using latency as a variable dependent on age and body height can be best used in practice when both reflexes are employed for demonstration of proximal pathology. The majority (69%) of 93 patients with various polyneuropathies showed abnormalities in both reflexes illustrating that proximal nerve segments are frequently involved. Four per cent had abnormal FCR H-reflexes with normal soleus H-reflexes whereas the reverse was found in 19% of the patients. Abnormal FCR H-reflexes occurred with normal motor and sensory conduction velocities in the peripheral part of the median nerve in 14%, whereas the reverse was seen in 12%, indicating that FCR H-reflex examination is a valuable supplement to conventional conduction studies for detection of electrophysiologically existing pathology. PMID- 3035103 TI - Congenital Lambert-Eaton myasthenic syndrome. AB - A 4 year old girl had been hypotonic and areflexic since birth with delayed milestones in motor development. Repetitive stimulation at high rates performed at 3 years elicited an incremental response typical of the Lambert-Eaton Syndrome. PMID- 3035104 TI - Transient uniocular visual loss on deviation of the eye in association with intraorbital tumours. AB - Five patients with unilateral orbital tumours are described in whom transient loss of vision occurred on deviation of the affected eye from the primary position. Other presenting features were diplopia, proptosis, poor visual acuity, visual field defects, pupillary abnormalities, fundal changes and altered colour vision. Abnormalities on fluorescein angiography suggest that the visual loss is due to transient ischaemia. Temporary uniocular loss of vision on eye movement may be an early sign of an intra-orbital mass. PMID- 3035106 TI - Neuralgic amyotrophy due to parvovirus infection. PMID- 3035105 TI - Segmental rigidity and spinal myoclonus as a paraneoplastic syndrome. AB - A 68 year old woman is described with persisting muscular rigidity of the left lower leg together with transient myoclonic jerking in the left quadriceps muscle. Six weeks after onset a small cell carcinoma of the lung became manifest. With radiotherapy and chemotherapy complete remission was achieved. Segmental muscular spasm improved at the same time. Necropsy revealed loss and degeneration of alpha-motor neuron cells at one side of the anterior horn of the lumbar enlargement. This case may represent another manifestation of paraneoplastic subacute motor neuronopathy. PMID- 3035108 TI - Demyelination in association with delayed type hypersensitivity. PMID- 3035107 TI - CSF protein and cellular profiles in various stages of HIV infection related to neurological manifestations. AB - CSF protein and cellular profiles were studied in 28 HIV-infected patients. Twenty of them had neurological complaints, but only 6 patients had objective neurological deficits such as dementia, ocular motility disorders or polyneuropathy. The serum/CSF HIV antibody ratio was on average lowest in acquired immunodeficiency syndrome (AIDS) (4 patients) and highest or almost normal in lymphadenopathy syndrome (LAS) (11) and asymptomatic seropositivity (ASX) (7), while it varied between these extremes in AIDS-related complex (ARC) (6). However, low values of the ratio were also found in the HIV-infected patients free of neurological symptoms and even in one ASX patient. The CSF IgG index was elevated in all these 4 general stages of HIV infection without any significant differences between them. The CSF/serum albumin ratio was slightly increased in patients with neurological deficits, but this ratio showed no association with any other clinical factor analysed. CSF leucocytes were increased in the early stages of the disease, but later the cellular reaction subsided. HIV was isolated from post mortem brain tissue of two AIDS patients and from the CSF of one of them. The results suggest increased intrathecal virus specific IgG synthesis, not only in patients with neurological deficits and at advanced stages of infection, but also in neurologically symptom-free subjects and at early infection. The lack of correlation between the increased virus specific IgG synthesis within the CNS and the presence of neurological symptoms suggests that neurologically "silent" areas of brain white matter are often affected in HIV infection. PMID- 3035109 TI - Effects of changes in tonicity of the extracellular solution on the size of vesicles in frog motor nerve terminals. AB - Frog nerve-muscle preparations were soaked and then fixed in solutions roughly isotonic to frog plasma, or in solutions that were markedly hypertonic or hypotonic. The hypertonic solution decreased the cross-sectional area of the muscle fibers but not of the synaptic vesicles. The hypotonic solution increased the cross-sectional area of the muscle fibres but did not produce comparable increases in the areas of the synaptic vesicles. Apparently the vesicles in situ do not behave as simple osmometers. This fact is significant for theories of exocytosis and for the mechanism of transmitter packaging in the vesicles. PMID- 3035111 TI - Noradrenaline-induced afterdepolarization in cat sympathetic preganglionic neurons in vitro. AB - Sympathetic preganglionic neurons of the intermediolateral nucleus were identified by antidromic stimulation in the slice of the T2 or T3 segment of the cat spinal cord. In normal Krebs solution, the action potential of these neurons had a shoulder on the repolarization phase and was followed by a long-lasting afterhyperpolarization (AHP). The AHP had a fast and a slow component. Superfusion of the slice with noradrenaline (NA), 10-50 microM, resulted in depression of the shoulder on the repolarization phase of the action potential, in the appearance of an afterdepolarization (ADP), which was absent in control conditions, and in depression of the slow component of the AHP. These effects were present whether the membrane potential of the sympathetic preganglionic neurons was decreased, increased, or not changed by NA. A typical ADP had time to peak of 50 ms and decay time of 200-500 ms; the amplitude was variable and large ADPs could be suprathreshold, causing repetitive firing. The amplitude and duration of the ADP increased with NA concentration. The appearance of the ADP seemed to be independent of the depressant effect of NA on the slow AHP. The ADP was associated with a decrease in neuron input resistance and was voltage dependent, being depressed in nonlinear fashion by membrane hyperpolarization. The ADP decreased in amplitude or disappeared within a range of membrane potentials from -70 to -90 mV. The ADP was reversibly suppressed by the Ca channel blocker cobalt (2 mM), by low Ca Krebs (0.25 mM), and by iontophoretic injection of ethyleneglycol-bis(B-aminoethyl-ether)-N,N'-tetraacetic acid into the cell. Increasing Ca concentration from 2.5 to 10.0 mM had no effect. The ADP was unaffected by tetrodotoxin, at a concentration blocking the Na spike, but was suppressed in Na-free medium, even when the Ca spike was prolonged by tetraethylammonium 20 mM. Changes in external K concentration from 3.6 to 2.5 or 10.0 mM did not change the ADP. Increasing intracellular Cl concentration or decreasing extracellular Cl concentration had no effect on the ADP. It is concluded that the ADP, evoked by NA, is due to an increase in membrane conductance involving Na and Ca ions, possibly a Ca-activated Na conductance. The ADP provides a mechanism with which NA may modulate sympathetic preganglionic neuron responsiveness to excitatory synaptic inputs. PMID- 3035110 TI - Colony formation in vitro as a prognostic indicator for primary breast cancer. AB - Cells from some, but not all, tumor biopsy samples form colonies when cultured in semi-solid media. The possibility that colony formation by progenitor cells in these tumors may reflect a more "aggressive" phenotype bearing clinical implications was examined in a series of 61 patients with primary breast cancer. Tumor cells from 32 samples formed colonies in vitro. There was no correlation between colony formation and any of the standard clinical parameters such as tumor size, nodal status, metastatic spread, or hormone receptor levels. Eighteen patients had inflammatory, locally advanced and/or detectable metastatic breast cancer at the time of surgery. Sixteen of these patients have progressed and 15 have died, with no relationship between colony formation and survival. For the 43 remaining patients, 23 had a tissue sample that gave rise to colonies in vitro; 14 of these have relapsed, with a median relapse-free survival (RFS) of 37.6 months, and eight have died with a median survival time of 46.8 months. This is compared with four relapses (median RFS not reached, P = .0043, Peto-Pike), and four deaths (median not reached, P = .1175) in the group without growth of the tumor specimen. These results indicate that colony formation is an independent prognostic parameter for breast cancer, which may be useful for selecting patients who would benefit from more intensive therapy. PMID- 3035113 TI - Pretreatment with hypertonic solutions increases quantal size at the frog neuromuscular junction. AB - Miniature end-plate potentials (MEPPs) are increased in size by pretreatment in a hypertonic Ringer. This paper is about the treatments that increase quantal size and the evidence that it occurs by packing more acetylcholine (ACh) in the quanta. Sartorius muscles were soaked for 2 h in a hypertonic Ringer, containing 200 mM NaCl in place of the usual 120 mM, and then returned to Ringer. In the hypertonic solution the miniature frequency was above 100/s. The treatment increased the size of the miniatures, recorded with an intracellular electrode after the return to usual Ringer, by a factor of approximately 2.5, compared with paired muscles kept in usual Ringer throughout. When the hypertonic solution was made with 200 mM sodium gluconate, after the return to usual Ringer, the miniatures increased in size by a factor of approximately 3.6. The large miniatures produced by the treatment fit a log normal probability distribution function, except for a variable small fraction of the largest events that may represent multiple releases. Miniatures recorded from untreated end plates fit better to a log normal distribution function than to a normal distribution function. The effects of the hypertonic treatments on the acetylcholine receptor (AChR) were accessed by measuring, after the return to usual Ringer, ACh noise, alpha-bungarotoxin binding, and the reversal potential. None of these measurements revealed significant differences between experimental and control preparations. The increases occurred when neostigmine was present in the hypertonic and usual solutions and when it was not, so changes in acetylcholinesterase (AChE) do not appear to be involved. Both the MEPPs and miniature end-plate currents, recorded with the two electrode voltage clamp, increased in size, so the increase is not owing to an rise in the input resistance of the muscle fiber. The increase in quantal size was prevented by including 1-10 microM AH5183 in the hypertonic solution. This drug blocks ACh uptake into synaptic vesicles. Therefore, it seems likely that the treatments act by increasing the quantity of ACh/quantum. Cyclohexamide (10 micrograms/ml), an inhibitor of protein synthesis, did not affect the increase in size caused by hypertonic solutions. Size increases were detected after 15 min in hypertonic solution. Most of the increase was complete after 1 h and there was no further increase after 2 h.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3035112 TI - Synaptic integration in excitatory and inhibitory crayfish motoneurons. AB - The passive integrative properties of two crayfish abdominal motoneurons, the fast flexor inhibitor (FI) and a posterior, ipsilateral fast flexor excitor (FE), were studied electrophysiologically and through simulations with multicompartment models of their electrotonic structures. Responses of the models to simulated giant neuron input were quite similar to the motoneurons' responses to giant neuron stimulation, which suggests that differences in the electrotonic structures and the sites of synaptic input to the two cells can account in large part for differences in their responses to a common input. A full action potential created in the initial axon compartment of the FI model produced attenuated potentials in the adjacent integrating segment compartment and contralateral soma compartment. These potentials are similar in amplitude and time course to attenuated antidromic action potentials recorded in the corresponding regions of the FI neuron. A location of the spike initiation zone of the FI at the initial axon segment is consistent with this result. The responses of FI to ipsi- and contralateral inputs are different. Shock of a single abdominal second root produced a larger, faster rising excitatory postsynaptic potential in the ipsilateral FI soma than in the contralateral soma. Second root shock also caused the contralateral FI to produce an action potential either alone or before the ipsilateral FI neuron. Responses of the FI model to ipsilateral and contralateral inputs differ in the same way as the cell's responses. Inputs to the FI model that are ipsilateral to the soma compartment produce larger responses there than do contralateral inputs. Conversely, those contralateral inputs produce larger responses in the initial axon compartment than do ipsilateral inputs. This difference results from the long integrating segment that connects the soma compartment to the initial axon compartment. These results can account for the FI responses to lateralized inputs. Unlike the responses of FIs, the soma responses of contralaterally homologous FEs to ipsilateral and contralateral second root shocks were similar in waveform and amplitude, with the ipsilateral root producing the larger response. This result is consistent with theoretical results from the FE model simulations. We conclude that a smaller size, larger input resistance and shorter membrane time constant allow the FE to respond to giant neuron input before the FI, and so help to achieve the proper timing of flexor contraction and relaxation during a tailflip.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3035115 TI - Estimation of portal-systemic shunting by rectal infusion of radiotracer. PMID- 3035114 TI - Iodine-131 MIBG scintigraphy of neuroendocrine tumors other than pheochromocytoma and neuroblastoma. AB - Metaiodobenzylguanidine (MIBG) locates most pheochromocytomas and neuroblastomas. The tracer is concentrated in intracellular storage vesicles by an active process. Many other neuroendocrine tumors of the amine precursor uptake and decarboxylation (APUD) series have hormonal storage vesicles and, thus, the potential to take up [131I]MIBG. A variety of neuroendocrine tumors in 57 patients were studied 1, 2, and 3 days after 0.5 mCi [131I]MIBG. Views from skull to pelvis were obtained. Results of MIBG scans were compared with all available imaging modalities (including plain radiography, liver scan, ultrasound, computed tomography, and angiography) and surgical exploration. The neuroendocrine nature of the tumor was determined by histology, immunohistochemistry, electron microscopy, and the assay of appropriate biogenic amines and peptide hormones. Results were (positive/total cases): carcinoids (four of ten), nonsecreting paragangliomas (three of three), sporadic medullary carcinomas of the thyroid (MCT) (one of five), familial MCT (one of 26), chemodectomas (two of five), oat cell carcinomas (zero of four), choriocarcinoma (one of one), atypical schwannoma (with storage granules) (one of one), Merkel cell skin cancer (one of one), islet cell carcinoma (zero of one). We conclude that a wide range of neuroendocrine tumors show [131I]MIBG uptake; tumors other than pheochromocytomas and neuroblastomas are less often seen scintigraphically, but in certain cases (e.g., carcinoid and nonsecreting paragangliomas) scintigraphy may be useful in depicting the extent and location of disease and may indicate therapeutic potential. Iodine-131 MIBG shows promise in the diagnosis and staging of tumors of varied types. PMID- 3035116 TI - Effects of a training model on active listening skills of post-RN students. AB - This study investigated the effects of a training module on the active listening skills of non-degree registered nurses. Active listening skills were defined as understanding what another person is saying and feeling and then communicating this understanding of his thoughts and feelings back to him. The sample consisted of 26 post-diploma RNs registered in the first year of a baccalaureate degree nursing program. Pretraining and posttraining data were collected when subjects verbally responded to two portions of the Behavioral Test of Interpersonal Skills for Health Professionals (BTIS). Subjects were audiotaped while responding and tapes were scored using BTIS guidelines. Paired t-tests were used to determine differences between pretraining and posttraining scores. Active listening scores increased significantly (p less than .0005) while attempts to suppress or discount speakers' feelings decreased significantly (p less than .005). A six hour training session significantly increased active listening skills. PMID- 3035117 TI - Strategy for teaching evaluation research in psychiatric/mental health nursing. AB - The evaluation research course presented in this paper is one strategy for introducing needed evaluation research content at the graduate level in nursing. The course was developed for students who were in their final semester of completing requirements for an MSN degree with a major in psychiatric/mental health nursing. To provide for active involvement in the evaluation research process, students evaluated a psychotherapy group in which they were therapists. The theoretical framework for both class content and the evaluation research project was based on Bloch's extension of Donabedian's approach to evaluation. Using the theoretical framework as a guide, students investigated both process and outcome as well as the relationship between process and outcome. Students reported that the knowledge and skills gained were valuable to their clinical practice. On the other hand, students found the course to be very demanding and difficult to complete in a 15 week period. One unexpected benefit, however, was that the course helped students to understand group process better. PMID- 3035118 TI - The effect of a course in nursing ethics on the relationship between ethical choice and ethical action in baccalaureate nursing students. AB - This study examined the effect of an ethics course on the variables of ethical choice and ethical action in baccalaureate nursing students. Data were collected from 17 students who were enrolled in a three-credit elective course in ethics and from 20 students who were matched for placement in the curriculum but who were not enrolled in the ethics course. The ethics course consisted of study of the major ethical principles and theories as well as guided ethical case analysis with emphasis on the role and responsibilities of the nurse. Ethical choice and ethical action were measured by Ketefian's Judgment About Nursing Decisions. The variables of moral choice and moral action were positively correlated r = .87 (p less than .001) in the students who were enrolled in the ethics course and they were negatively correlated at r = -.32 (p .34) in the students who were not enrolled in the ethics course. Overall scores of the two groups on both column A (Ethical Choice) and column B (Ethical Action) were not significantly different. The total scores of the ethics group were higher. The results of this study lend support to the inclusion of a course in ethics in nursing curricula. Continued research into this area should continue to be a high priority. PMID- 3035119 TI - Case studies: an alternative learning/teaching method in nursing. PMID- 3035121 TI - Practice: a sanctioned faculty role. PMID- 3035123 TI - Campus day care centers: a unique opportunity for learning. PMID- 3035122 TI - Summer specialty camps: an overlooked site for student nurses' clinical placement. PMID- 3035120 TI - Behaviorally based clinical evaluation. AB - For the past two years this clinical evaluation tool has been revised biannually by the original committee. Feedback has been solicited from the entire faculty and incorporated into the subsequent revisions. In using the tool it is apparent that there are benefits for both students and faculty. Evaluation of the student is simplified because it is based on behaviorally described competencies. Because of this, consistency is further assured regardless of the number of evaluators. The faculty not only have a tool that is short and manageable enough to provide daily feedback but they also have the ease of using pass or fail for most of the evaluations. Students are better able to channel their energies and ultimately their behaviors to focus on necessary competencies to become an effective care giver. Students have the benefit of objective evaluation of their behaviors with an opportunity for daily feedback. The student is aware of expected performance levels prior to evaluation as well as the need for progress in those competencies. While the tool in its present format still has some shortcomings, the entire faculty have had the pleasure of knowing that as a group they have created a workable and discriminating evaluation tool. PMID- 3035124 TI - Relationship of anxiety level and recall of information to the interval between orientation and first patient care day. AB - Sophomore students' responses to two different intervals between orientation and first clinical day in medical/surgical nursing were examined. Variables studied were anxiety state and ability to recall information which had been presented. Prior to orientation all students were given the Spielberger STAI-S and STAI-T anxiety scales. At the beginning of the first laboratory session the students completed the STAI-S scale and a teacher-made quiz on information presented in orientation. Students in Group I (n = 51) had their first laboratory one week after orientation. Group II (n = 39) had their first laboratory the day following orientation. There was no significant difference between the two groups on cumulative grade point average, beginning STAI-T or STAI-S. Likewise, there was no significant difference between the scores made on the quiz. (The validity and reliability of the quiz were not determined.) There was a significant difference between the two groups on changes in state anxiety from orientation to the first laboratory. Group I (laboratory one week after orientation) was more anxious than Group II (laboratory one day after orientation) (p = 0.024). Both groups were significantly more anxious in the Spring than Fall quarter (p = 0.034). It was concluded that anxiety related to a new clinical experience might be lessened if the first clinical day closely followed orientation. It was also noted that students may be more anxious in the Spring than in the Fall of the year. PMID- 3035126 TI - Ten criteria for evaluating qualitative research proposals. AB - With the proliferation of interest in qualitative research in nursing comes the attendant problem of how to evaluate it appropriately. Qualitative research has its own unique history, philosophical foundations, and methodologies that separate it from the quantitative approach. Although the literature is crowded with guidelines for evaluating the latter, little is offered for the qualitative reviewer. The Research Proposal Evaluation Form: Qualitative Methodology is a partial solution to this dilemma. It provides a framework for critiquing the proposal phase of a qualitative study and can be an important guide both for the educator and for the novice researcher. PMID- 3035125 TI - Training on selected self-management techniques and the generalization and maintenance of interpersonal skills for registered nurse students. AB - This study examined the effects of training on selected self-management procedures (post-workshop practice, self-monitoring, self-assessment, and self reinforcement) on the generalization and maintenance of interpersonal skills with a group of registered nurse students. The findings indicate that those self management procedures enabled the student nurses to maintain a positive attitude toward the use of interpersonal skills in their work and maintain their skill level over a period of time significantly better than a control group who received interpersonal skills training only. No significant differences were observed between the two groups as to their ability to maintain their knowledge of the concepts taught during training or their ability to generalize their skills to the hospital ward immediately following training. PMID- 3035128 TI - Nursing students' perceptions of clinical experience. AB - Senior nursing students were interviewed in this study to better understand the clinical learning experience from the students' point of view. Results of the study revealed that the nursing students were indeed learning in their clinical experience. The major categories of learning were classified as nursing skills, time management, and professional socialization. The quality of learning was reportedly affected by the quality of the student's preparation, characteristics of the instructor, and the variety of clinical opportunities to which students were exposed. The data also reflected a pattern of student development which was separated into three stages. The first stage was permeated with anxiety and obsession with the rules of task performance. The second stage was a difficult transition period where students struggled with identifying the roles of nurses. During the final stage, the students become more comfortable with performing nursing tasks and become interested in expanding their role and becoming more independent. As the students strived for independence, they identified more closely with staff nurses and withdrew from instructors. PMID- 3035127 TI - Social problems encountered by public health nurses: identification and response differences according to education and experience. AB - Do nurses differ in relation to the identification of social problems and actions taken according to educational preparation? Using data from a larger field study, baccalaureate nurses (BSNs) were compared to Non-BSNs. The two groups did not differ in the number of problems that were identified, but BSNs did identify more problems related to non-immediate needs. The nurses took up to ten actions (means = 4.6) per problem, and the average number per BSN was less than the average per Non-BSN. The data lend some support to the position that BSNs are more likely to be self-assured and self-directed. Several questions are raised that are relevant to educators and administrators. PMID- 3035129 TI - Spiritual care: integration into a collegiate nursing curriculum. PMID- 3035131 TI - Adult education: its implications for baccalaureate nursing education. PMID- 3035130 TI - Evaluating the use of a data base system with community health nursing students. AB - The Data Base Management System for community diagnosis was used during two academic semesters with 80 baccalaureate nursing students. At the end of the two semesters the DBMS was evaluated from feedback obtained through faculty discussions, final student papers and oral presentations. The DBMS met the goals established to evaluate the tool. The clarification of categories for essential data is an ongoing process. PMID- 3035133 TI - Recruitment strategy for increasing minority enrollment in nursing and other professional schools. PMID- 3035132 TI - Utilization of board gaming for conceptual models of nursing. AB - Gaming can be an effective teaching strategy for reinforcing and motivating students to learn. Development of the game discussed in this article is not intended to replace classroom instruction. Instead, it is intended to augment course content. Board gaming can make learning a pleasant experience. It provides students and educators an opportunity to interact in an informal atmosphere and provides enjoyment, while at the same time may promote learning. Therefore, educators should consider further development of games for use with students in nursing education. PMID- 3035134 TI - Institutional goal analysis: an approach to program evaluation. AB - Although nursing literature discusses the importance of goals in nursing, little if any research has been conducted on the subject. Many of the studies focusing on nursing education deal with faculty turnover, effectiveness of various types of nursing programs, and the role and functions of the nurse. To better understand these problems, an understanding of the current goals of various nursing programs is needed. This can establish a basis on which to explore other problems in nursing education and the nursing profession. Conducting an analysis of goals on a nursing program not only contributes to the profession of nursing but also benefits the individual schools by revealing areas of goal incongruency within each program. The use of the Institutional Goals Inventory is recommended for evaluation of all nursing programs. It can be useful not only to baccalaureate educators, but also to those involved in associate degree and diploma education as well as graduate nursing education. After identifying areas of goal consensus and differences, a delphi approach can be utilized to solve problems and establish relevant and realistic goals. PMID- 3035135 TI - The student health center: a unique clinical experience for nursing leadership students. PMID- 3035136 TI - Coping with anxiety and tension. PMID- 3035137 TI - Tenure and promotion: an update on university nursing faculty. AB - The status of tenure and promotion practices in nursing schools in university settings was studied. Forty nursing administrators from nursing programs around the country responded to a questionnaire regarding requirements to achieve tenure and/or promotion in the areas of teaching, research and scholarly activities, clinical competence, and community service. Findings revealed that while a growing number of nursing programs were more integrated into the academic community, many programs continued to struggle with the need for more faculty prepared at the doctoral level and for increased research activities. PMID- 3035138 TI - Combining community health and psychosocial nursing: a clinical experience with the homeless for generic baccalaureate students. AB - A report is made of a teaching approach which combined community health and psychosocial nursing clinical experiences for senior level, generic, baccalaureate nursing students over a period of two academic quarters. An urban community center and shelter for the homeless was the setting for this unique clinical approach. Areas on which the evaluation of the clinical experience focused included the clinical design, student activities, types of clients seen, and student and faculty views. Strengths and limitations of the experience were identified. The progression of the student group in adapting to the homeless population was described in detail. Faculty concluded that by sharing their clinical expertise in this combination clinical setting with the homeless, generic students were able to maximize and coordinate their academic and clinical activities. The homeless setting is demonstrated to be an excellent arena for student learning when faculty provide structure, support, and guidance. PMID- 3035140 TI - Developing an evening clinical experience for baccalaureate community health nursing students. AB - With the ever-changing directions in health care delivery, baccalaureate nursing instructors are being challenged with the task of seeking out innovative approaches to community health nursing clinical experiences. With the focus on "community as client," an evening clinical program was designed and piloted for nontraditional nursing students who, because of daytime employment, were in need of evening courses to further their nursing education. The pilot project incorporated family nursing care, community-centered practice, and observational experiences. The evening community health nursing clinical experience was found to be mutually beneficial to clients and students and served to fill a gap in health teaching within the community. PMID- 3035141 TI - Learning interdisciplinary and assessment skills through videotaped client interviews and collaborative planning. PMID- 3035139 TI - Communicating leveled clinical expectations to nursing students. AB - Nurse educators agree on the need for objectives in the classroom situation and frequently communicate them in a written form. Written expectations for clinical, however, are rarely given to student nurses prior to their clinical experience. Clinical expectations are understood by instructors, but communicating them to students result in mixed interpretations. The article describes the development of a written method of communicating clinical expectations to solve this problem. This method correlates clinical expectations with classroom content and course objectives and levels the expectations in increasing complexity by using the nursing process as a framework. The written leveled expectations are distributed to student nurses on a weekly basis. Clinical experiences are enhanced for students and faculty who use this method. PMID- 3035142 TI - Peer evaluation as a teaching-learning strategy in baccalaureate education for community health nursing. PMID- 3035143 TI - The need for cultural concepts in nursing curricula. PMID- 3035144 TI - Confirmation of qualitative research findings in the clinical setting: a strategy to promote research application among baccalaureate nursing students. PMID- 3035146 TI - Small group work: a strategy to promote active learning. PMID- 3035145 TI - Use of management and ethical case studies to improve decision-making skills in senior nursing students. PMID- 3035147 TI - Effects of dietary fiber, carbohydrate, lipid and protein levels on serum and liver lipids in rats. AB - Response-surface regression analysis was used to study dietary levels of fiber, carbohydrate, lipid and protein to minimize serum and liver cholesterol and triglyceride levels and maximize serum high-density lipoprotein-cholesterol levels of male weanling rats. Because the dietary components were not statistically independent, they were studied in combinations of three variables. The three-variable combinations were the most useful in locating the desired maximum or minimum lipid responses in terms of the proportions of the dietary components. These analyses indicated that dietary carbohydrate, lipid and protein were better than dietary fiber for predicting the serum and liver lipid response levels. Response-surface contours and three-dimensional plots were developed for each lipid response except serum triglycerides, which were not predictable. The contours and three-dimensional plots were used to help determine those combinations of the diet components that would produce the desired maximum or minimum lipid responses. The statistical analyses indicated that the desired lipid response levels could be attained with a diet consisting of 3-5% neutral detergent bran fiber, 6-10% lipid, 54-55% carbohydrate, 26-30% protein and 4.7% vitamins and minerals. PMID- 3035148 TI - Studies on the vitamin D endocrine system in the avian. AB - This report summarizes the current understanding of the mode of action of vitamin D and emphasizes the contributions to this system that have been based on results obtained in avian species. A complex endocrine system coordinates the metabolism of vitamin D into 1,25-dihydroxycholecalciferol[1,25(OH)2D3] and 24R,25 dihydroxycholecalciferol[24R,25)OH)2D3] and 28 other metabolites; of these 16 were originally isolated and chemically characterized from avian systems. Key advances in understanding the mode of action of the seco-steroid 1,25(OH)2D3 and the scope of its action have been made by studying the tissue distribution of both its receptor and its gene-induced product, a 28,000-dalton calcium-binding protein termed calbindin-D28K. To date no less than 23 tissues have been found to have specific 1,25(OH)2D3 receptors; of these 10 were identified in avian studies. Similarly, nine tissues express the vitamin D-induced calbindin; seven have been reported in avian tissues. The second dihydroxylated metabolite, 24R,25(OH)2D3, has been reported to be capable of inducing a variety of specific biological effects, some when the steroid is administered alone and some in the presence of its companion dihydroxylated metabolite 1,25(OH)2D3. There is emerging evidence for the existence of specific receptors for 24R,25(OH)2D3, particularly in chondrocytes and parathyroid glands (both studied in avian systems). These observations collectively demonstrate the broad scope of the vitamin D endocrine system. PMID- 3035149 TI - Importance of cereal phytase activity for phytate phosphorus utilization by growing pigs fed diets containing triticale or corn. AB - In contrast to corn, wheat and triticale exhibit high phytase activities. This enzyme enhances phytic phosphorus availability, as demonstrated in pigs given wheat diets. To study the utilization of triticale phosphorus in pigs, the importance of dietary phytase content and the mineral and bone disorders related to high phytate feeding, a nutritional experiment was carried out in 12 growing pigs fed either a corn- or a triticale-based diet for 6 wk. The diets were almost identical except for the cereal component; their phosphorus contents were low (0.4%) and mainly phytic. The following parameters were measured: calcium and phosphorus balances, bone and plasma contents of calcium and phosphorus, plasma vitamin D metabolites and parathyroid hormone (PTH), bone bending moments and intestinal phosphatase activities. Both diets provoked a phosphorus deficiency, but hypophosphatemia occurred less rapidly, hypercalciuria and hypophosphaturia were less marked and phosphorus availability was greater when the triticale diet was fed. This was attributed to the high phytase content of triticale because intestinal phytase and alkaline phosphatase activities were similar in pigs fed either diet. Calcium absorption was not modified by calcium retention was greater for pigs fed triticale and led to higher bone scores. In conclusion, the higher the phytase activity of the diet, the greater the phytate P availability and the lower the bone-mineral disorders. PMID- 3035151 TI - Effects of rice fiber on fecal weight, apparent digestibility of energy, nitrogen and fat, and degradation of neutral detergent fiber in young men. AB - The effects of rice fiber on fecal weight, transit time, frequency of defecations, digestibility of nutrients and blood status were investigated in 5 healthy young men. Each of them consumed a brown rice diet and then a polished rice diet for 2 weeks respectively. Both diets contained 1.2 g protein per kg body weight. The brown rice diet contained 2 times as much dietary fiber as the polished rice diet. When they consumed the brown rice diet, it showed an increase of fecal weight and decrease of digestibility of energy, nitrogen and fat. Nitrogen balance was not significantly different and kept zero balance on both diets. Concentration of plasma cholesterol was not significantly different. The results suggest that rice fiber produced an increase in fecal weight, which is assumed to be effective in preventing colonic disease in advanced countries and does not affect plasma lipid level. PMID- 3035150 TI - Virus-inactivating effect of D-isoascorbic acid. AB - The effect of D-isoascorbic acid, an epimer of L-ascorbic acid, on viruses was investigated using a wide variety of bacterial viruses (phages) as model systems. D-isoascorbic acid exerted an inactivating effect on all phages examined. The reaction mechanism of virus inactivation by D-isoascorbic acid was investigated using phage J1 as a model system. Bubbling oxygen through the reaction mixture and the addition of H2O2 or transition metal ions into the reaction mixture enhanced the phage inactivation by D-isoascorbic acid. In contrast, nitrogen bubbling and the addition of reducing agents, chelating agents or radical scavengers prevented phage inactivation. Experiments using specific radical scavengers, superoxide dismutase or catalase showed that OH. could be mainly responsible for phage inactivation by D-isoascorbic acid. These findings are similar to those obtained with L-ascorbic acid, and indicate that phage inactivating activity is independent of the stereoisomerism with inversion of the hydroxyl group at carbon 5 of ascorbic acids. PMID- 3035152 TI - Effects of flavonoids on xanthine oxidation as well as on cytochrome c reduction by milk xanthine oxidase. AB - That flavonoids inhibit xanthine oxidase from cow milk was confirmed by measuring oxygen consumption with an oxygen electrode. In contrast, flavonoids did not inhibit glucose oxidase, another oxygen consuming enzyme. Among the flavonoids tested, quercetin, kaempferol, myricetin, chrysin, quercitrin, and morin were potent inhibitors of xanthine oxidase; their inhibition rates (%) were 80, 70, 69, 62, 59, and 51 at 100 microM (except chrysin at 50 microM), respectively. The xanthine oxidase-inhibiting activities of the flavonoids were not always well correlated with the suppressive activities of the flavonoids on cytochrome c reduction by a xanthine-xanthine oxidase system. The inhibition of xanthine oxidase by quercetin was not affected by cupric ion. The partition rates of the flavonoids between n-butanol and a buffer solution seemed to account for some of the inhibition. PMID- 3035153 TI - Work status and response to epidural steroid injection. AB - A total of 187 patients with lumbar radiculopathy underwent epidural steroid injection. On follow-up 1 to 3 years later, patients were asked if their initial injury was work-related and if they were unemployed as a result of their injury. They also rated their response to injection. Fifty-two percent of the patients with work-related injury v 68% of the patients with non-work-related injury responded to epidural steroid injection (P = 0.46). PMID- 3035154 TI - Hydroxylapatite cranioplasty directly over dura. AB - Seven patients who had noticeable defects of their frontal bone were reconstructed with dense hydroxylapatite (HA) particles with or without autogenous bone placed directly over the dura. The results indicate that HA is well tolerated over dura; no meningitis occurred with follow-up of one to 3 1/2 years. The clinical response was excellent and complications were minor, generally related to particle control and settling. PMID- 3035155 TI - Ultrastructural characteristics of the interdigitating dendritic cell in dermatopathic lymphadenopathy of mycosis fungoides patients. AB - The ultrastructure of interdigitating dendritic cells in dermatopathic lymphadenopathy from patients with mycosis fungoides was investigated. The most remarkable findings were: a marked hypertrophy of the smooth endoplasmic reticulum with a peculiar concentric arrangements of the cisternae around lipid droplets, an abundance of cytoplasmic lipid droplets and the presence of paranuclear chromatoid-like bodies. It is suggested that these ultrastructural features may correspond to some unknown functional capability of the interdigitating dendritic cell. PMID- 3035156 TI - Acquired immune deficiency syndrome presenting as progressive infantile encephalopathy. PMID- 3035157 TI - Pneumocystis carinii pneumonia in infants given adrenocorticotropic hormone for infantile spasms. PMID- 3035158 TI - Fructose 1,6-bisphosphatase activity in two Trypanosoma cruzi morphological forms. PMID- 3035159 TI - Intraarterial injection of anti-tumor drugs dispersed in lipid contrast medium: a choice for initially unresectable hepatoblastoma in infants. AB - Intraarterial injection of anti-tumor drugs (THP-ADR, CDDP, 5-FU) dispersed in lipid contrast medium (Lipiodol, Laboratorie Guerbert, France) was used in an infant with initially unresectable hepatoblastoma. Lipiodol complex selectively accumulated in the tumor tissue and may keep the chemotherapeutic agents in the tumor tissue for a longer period of time, and a significant reduction of the tumor with a four-day half-life of alpha-fetoprotein (AFP) was followed immediately after the institution of chemotherapy. Successful resection of decreased tumor by extended right hepatectomy under more favorable conditions was performed 2 months after diagnosis. Intraarterial chemotherapy with Lipiodol was particularly useful in potentiating the cytoreduction of anti-tumor drugs and was also useful in reducing the toxicities of anti-tumor drugs to the host. PMID- 3035160 TI - Plasma immunoreactive beta-endorphin, ACTH and cortisol concentrations in mothers and their neonates immediately after delivery--their relationship to the duration of labor. AB - In 29 cases of vaginal delivery with normal outcome and 4 cases of cesarean section, the concentrations of beta-endorphin, ACTH and cortisol were determined in maternal venous and umbilical venous plasma immediately postpartum. According to duration of labor and mode of delivery the cases examined were classified into three groups: Group A (18 cases) = vaginal delivery of less than 10 hours' duration, Group B (11 cases) = vaginal delivery of more than 10 hours' duration of labor, Group C (4 cases) = cesarean section under general anesthesia. With the exception of one, the deliveries took place at term. The 33 neonates were in a very good clinical state 5 minutes after parturition (11 Saling points as median value). For measurement of the hormone concentrations radioimmunoassays were used. In Group a the mean beta-endorphin concentration in maternal plasma amounted to 150.9 +/- 16.3 pg/ml, that in neonatal plasma to 239.2 +/- 23.5 pg/ml (means +/- SEM). In Group B plasma beta-endorphin, both maternal and neonatal, was slightly higher than in Group A: 153.0 +/- 12.0 pg/ml (maternal) and 260.9 +/ 37.1 pg/ml (neonatal). The differences between maternal and neonatal beta endorphin levels were statistically significant: Group A p less than 0.01, Group B p less than 0.05; chi 2-test. The mean ACTH concentrations in the plasma of the newborn infants were also found to be considerably higher compared with those in the plasma of their mothers: Group A 78.2 +/- 16.5 pg/ml (maternal) and 98.0 +/- 23.3 pg/ml (neonatal); Group B 98.0 +/- 20.1 pg/ml (maternal) and 165.8 +/- 39.6 pg/ml (neonatal).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3035161 TI - Preparation and characterization of human gingival cells. PMID- 3035162 TI - The effect of chlorhexidine on blood cells. PMID- 3035163 TI - Demonstration of tissue collagenase activity in vivo and its relationship to inflammation severity in human gingiva. PMID- 3035164 TI - Salt and peroxide compared with conventional oral hygiene. I. Clinical results. AB - The purposes of this 2-year longitudinal study were to: compare the clinical effectiveness of patient applied sodium bicarbonate, hydrogen peroxide, and sodium chloride (S/P) to the use of conventional oral hygiene methods and to investigate the motivational effect of using phase-contrast microscopy in teaching effective oral hygiene. Initially, 972 subjects were screened for signs of periodontitis. From these, 347 with early to moderate periodontitis were selected and each was randomly assigned to one of four home treatment regimens after scaling and root planing. The four treatment regimens included: conventional oral hygiene procedures, conventional oral hygiene procedures plus phase-contrast demonstration of subgingival microbial forms for oral hygiene motivation, S/P oral hygiene, and S/P oral hygiene plus phase-contrast demonstration of subgingival microbial forms for oral hygiene motivation. Plaque, bleeding, gingival inflammation, probing depth, and clinical attachment level were recorded at baseline, 8, 16, and 24 months. Subjects were recalled for reinforcement of oral hygiene and periodontal prophylaxis at various intervals. Data were analyzed based on disease severity, location of index sites and compliance. The results indicated that both conventional oral hygiene procedures and the S/P regimen were effective in reducing clinical signs of disease when combined with professional care. There were no differences between the two regimens in clinical effectiveness and trends favoring microscopic viewing of subgingival plaque for motivational purposes were not statistically significant. PMID- 3035165 TI - Salt and peroxide compared with conventional oral hygiene. II. Microbial results. AB - This study was designed to investigate the effect of conventional oral hygiene (n = 116 subjects) versus a salt and peroxide oral hygiene regimen (n = 115 subjects) on subgingival microorganisms. Subgingival plaque for microscopic evaluation was obtained from eight index tooth sites in each of 231 adult subjects. Microbial forms were microscopically identified at baseline, 8, 16, and 24 months. For both oral hygiene groups, cocci were increased (P less than 0.05) and motile rods were decreased (P less than 0.05) at 8 months and returned to baseline by 16 months. Spirochetes were decreased (P less than 0.05) and remained low through 24 months in both oral hygiene groups. The frequency of agreement between clinical (bleeding) and microbial (greater than or equal to 15% spirochetes or motile rods or greater than or equal to 20% spirochetes + motile rods) criteria for instrumentation was 59.8%. It was also found that fewer total instrumentations for test subjects were observed when microbiological criteria were used as compared with clinical criteria. The greater number of instrumentations based on clinical criteria was highly significant (P less than or equal to 0.001). A significant change in microbial signs associated with peridontal disease may be obtained with either a conventional oral hygiene or a salt and peroxide oral hygiene home care regimen. PMID- 3035167 TI - [Purification and determination of core protein p24 of human T-cell leukemia virus using monoclonal antibody]. PMID- 3035166 TI - Salt and peroxide compared with conventional oral hygiene. III. Patient compliance and acceptance. AB - This study was undertaken to evaluate patient compliance with, and acceptance of, a salt and peroxide oral hygiene regimen compared with conventional oral hygiene regimens without or with the use of phase-contrast microscope viewing of subgingival plaque over a period of 2 years. A total of 231 subjects with early to moderate periodontitis were randomly divided into four groups. All groups were repeatedly instructed and motivated in their respective regimens. Subjects also received scaling and root planing using clinical and microbial criteria. Compliance with, and acceptance of, the two oral hygiene regimens were determined at the end of the study using a structured self-administered questionnaire. Results indicated that 74% and 58% (P less than or equal to 0.01) of subjects in the conventional and salt/peroxide groups, respectively, used their assigned regimen 4 to 7 days a week during the entire study. More than half of the subjects (54%) using each of the oral hygiene regimens indicated that they flossed once daily. Inconvenience was cited by 23% of the conventional and 43% of the salt/peroxide groups (P less than or equal to 0.01) as the main reason for not using their regimens. Twenty-three per cent of conventional group and 14% of salt/peroxide group indicated that shared their oral hygiene supplies with others. Eighty per cent and 57% (P less than or equal to 0.01) of the conventional and salt/peroxide groups, respectively, stated that they liked their regimens. Ninety-six per cent of all subjects felt that their regimen helped their periodontal status.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3035168 TI - Restriction enzyme map of cryptic plasmid accompanying pR711b and pR409; identity of pR409 and pR388. AB - A reference strain containing the IncW plasmid, pR409, was found to contain also a cryptic, conjugative plasmid, here named pJR15. After the separation of the two plasmids into different strains of bacteria, pR409 was found to have an identical restriction enzyme map to the IncW plasmid, pR388. Both pR409 and pR388 were isolated from the same hospital in London and confer sulphonamide and trimethoprim resistance. Originally pR409 was reported to confer resistance to tetracycline, but this was not confirmed by the Plasmid Reference Center. pR409 appears to represent another isolate of pR388 in a different host background. The restriction enzyme map of the cryptic plasmid, pJR15 (39 kb), has been determined. pJR15 was found compatible with plasmids from 15 different incompatibility groups and has been found also in the reference strain of the IncD plasmid, pR711b. The HindIII and BglII digests of pR711b are shown. The possible presence of the conjugative plasmid, pJR15, should be examined for studies on the sex pili or chromosomal mobilization properties of pR711b. PMID- 3035169 TI - Antihypertensive 1-acyl-4-[2-(1,4-benzodioxan-2-yl)-2-hydroxyethylamino]pip eridines. AB - Several 1-acyl-4-[2-(1,4-benzodioxan-2-yl)-2-hydroxy-ethylamino]piperidine s were prepared and a number of the compounds showed antihypertensive activity in the spontaneously hypertensive rat (SHR). This activity was specific for the (2S, 2R) enantiomers. General pharmacological evaluation and ligand binding data on selected compounds indicated a moderate degree of alpha 1- and beta-antagonistic activity. The alpha 1 antagonism was probably not of sufficient magnitude to explain the blood pressure lowering activity in the SHR. PMID- 3035170 TI - Enzyme immunoassay discrimination of a new angiotensin-converting enzyme (ACE) inhibitor, cilazapril, and its active metabolite. AB - Simple and sensitive enzyme immunoassays (EIAs), discriminating a new angiotensin converting enzyme (ACE) inhibitor, cilazapril [9(s) - [1(s)-(ethoxycarbonyl)-3 phenylpropylamino]-octahydro-10-oxo-6H- pyridazo[1,2-a] [1,2]diazepine 1(s)carboxylic acid] and its active metabolite [9(s)-[1(s)-carboxy-3 phenylpropylamino]-octahydro-10-oxo-6H- pyridazo[1,2-a][1,2]diazepine 1(s)carboxylic acid] were developed for pharmacokinetic studies of this drug which is used as an antihypertensive agent. These assays can be performed directly on serum or plasma specimens without pretreatment. The EIA for cilazapril (prodrug) allowed the determination of as little as 30 pg/mL, while a 1000-times greater concentration of its active metabolite was required to achieve the same extent of inhibition. The EIA for the active metabolite exhibited a sensitivity and specificity similar to those of the prodrug. Plasma specimens from a human volunteer study and a marmoset subacute toxicity study were assayed by these two newly developed EIAs. The plasma levels of active metabolite determined by the EIA were compared with those assayed by radioenzymatic assay, and a good correlation was observed between them. PMID- 3035171 TI - Determination of 7-piperidino-1,2,3,5-tetrahydroimidazo[2,1-b]quinazolin-2-one (DN-9693) in human plasma and urine by high-performance liquid chromatography with electrochemical detection: application to studies on disposition of DN-9693 in healthy human volunteers. AB - A procedure for the determination of a newly developed antiadhesive agent, 7 piperidino-1,2,3,5-tetrahydroimidazo[2,1-b]quinazolin-2-on e (DN-9693; 1), in human plasma and urine at low nanogram levels by high-performance liquid chromatography with electrochemical detection is described. Plasma and urine samples were made alkaline with 1 M KOH solution and extracted with chloroform. After removing organic solvent, the extracts reconstituted in methanol were applied to a reversed-phase octadesyl-packed column and eluted with a methanol: phosphate buffer (pH 8.0) mobile phase. The applied potentials were set at 0.80 and 0.95 V versus a silver-silver chloride reference electrode for the analyses of plasma and urine specimens, respectively. The method provides a recovery of greater than 90% and a within-day precision of less than 5%. The minimum quantifiable levels for 1 in plasma and urine using 1 mL of sample are 1 and 4 ng/mL, respectively. The method has been successfully used for the determination of 1 in plasma and urine samples from healthy human volunteers after intravenous infusion of the drug. PMID- 3035172 TI - 2,5-Dimethyl-2'-hydroxy-9 alpha- and 9 beta-(3-methylbutyl)-6,7-benzomorphans and N-substituted compounds in the 9 alpha-(3-methylbutyl) series: chemistry, pharmacology, and biochemistry. AB - 2,5-Dimethyl-2'-hydroxy-9 alpha-(3-methylbutyl)-6,7-benzomorphan, the 9 beta analogue, and 9 alpha-N-substituted (N-ethyl, propyl, butyl, pentyl, hexyl, phenylethyl, allyl, and cyclopropylmethyl) compounds were synthesized and evaluated biochemically and pharmacologically. The 9 beta N-methyl compound was found to be as potent as morphine in the mouse hot plate assay and had one seventh the affinity of morphine for the opioid receptor. The N-alkyl and N phenethyl 9 alpha-substituted compounds were either inactive or relatively ineffective as antinociceptive agents. None of the examined compounds substituted for morphine in single-dose suppression studies in the rhesus monkey. The N cyclopropylmethyl compound in the 9 alpha series had half the narcotic antagonist potency of nalorphine and one-eighth of its affinity for the opioid receptor. The 9 alpha-(3-methylbutyl) moiety, unlike bulky substituents in the 9 beta position of 6,7-benzomorphans, generally lowers affinity for the mu opioid receptor and diminishes their in vivo activity as agonists or antagonists. PMID- 3035173 TI - [Actions of acetylcholine on vascular smooth muscles]. PMID- 3035174 TI - Platelet alpha 2-adrenoceptor binding and function during the menstrual cycle. AB - Blood platelet receptors are widely used as peripheral models of central nervous system receptors, particularly in attempts to understand the biological basis of a number of neurological and psychiatric disorders. It is important to differentiate factors other than the primary disease process which may influence platelet receptors. One such potential factor is the menstrual cycle. In this study we have determined platelet high-affinity alpha 2-adrenoceptor binding, using the agonist ligand 3H-UK-14,304 and platelet aggregatory responses to adrenaline in 14 healthy young women, sampled on four occasions at weekly intervals. Our results indicate that, within the limits of individual variation, neither the KD or Bmax of high-affinity 3H-UK-14,304 binding or the aggregatory responses to adrenaline differed significantly between various stages of the menstrual cycle. PMID- 3035175 TI - Granular cell myoblastoma of the cystic duct: a case report and review of the literature. PMID- 3035176 TI - [Pseudolinitis plastica of breast origin]. AB - The most common type of gastric metastases secondary to carcinoma of the breast is the one showing a linitis plastica appearance. The roentgenologic aspect when performing an upper gastrointestinal examination with barium contrast is typical and allows the differential diagnosis with the primitive linitis. The linitis plastica type of metastatic breast carcinoma results from transmural infiltration of the stomach wall with initial deposit of tumor cells in the submucosal or subserosal regions. CT provide valuable information concerning the metastatic spread. PMID- 3035177 TI - Nonequilibrium opioid antagonist activity of 6,14-dideoxynaltrexone derivatives. AB - A series of 6,14-dideoxynaltrexones that contain different electrophiles in the 6 position were synthesized and evaluated for nonequilibrium opioid antagonist activity in the guinea pig ileum and mouse vas deferens preparations. Members 3-5 of the series possessed irreversible antagonist activity profiles similar to those previously reported for the 14-hydroxy analogues. In contrast, the 14-deoxy beta-funaltrexamine (14-deoxy-beta-FNA) analogue (6) exhibited a profile of irreversible antagonist activity that differed from that of beta-FNA. It was concluded that the 14-hydroxy group is not essential for irreversible blockage when the electrophile is capable of reacting with a broad spectrum of nucleophiles. However, with a highly selective electrophile such as the fumarate group, the 14-hydroxy function appears to play a role in aligning the molecule to optimize attack by a receptor-based nucleophile. PMID- 3035178 TI - Synthesis and antiviral evaluation of carbocyclic analogues of 2-amino-6 substituted-purine 3'-deoxyribofuranosides. AB - Carbocyclic analogues of 2-amino-6-substituted-purine 3'-deoxyribofuranosides were synthesized by beginning with (+/-)-(1 alpha,3 alpha,4 beta)-3-amino-4 hydroxycyclopentanemethanol and 2-amino-4,6-dichloropyrimidine. The route parallels the earlier syntheses of the corresponding ribofuranoside and 2' deoxyribofuranoside analogues. The 2-amino-6-chloropurine, guanine, and 2,6 diaminopurine derivatives and the analogous 8-azapurines were prepared. The analogue (3'-CDG) of 3'-deoxyguanosine is active in vitro against a strain of type 1 herpes simplex virus (HSV-1) that induces thymidine kinase and is modestly active against a thymidine kinase inducing strain of type 2 HSV. 3'-CDG is not active against a strain of HSV-1 that lacks the thymidine kinase inducing capacity, whereas the carbocyclic analogue of 2-amino-6-chloropurine 3' deoxyribofuranoside is active against that strain. The carbocyclic analogue of 2,6-diaminopurine 3'-deoxyribofuranoside displayed modest activity in vitro against influenza virus. PMID- 3035180 TI - Synthesis and structure-activity relationships of potent new angiotensin converting enzyme inhibitors containing saturated bicyclic amino acids. AB - The synthesis of a series of novel, potent angiotensin converting enzyme (ACE) inhibitors containing saturated bicyclic amino acids in place of proline is described. Octahydroindole-2-carboxylic acid, octahydroisoindole-1-carboxylic acid, and octahydro-3-oxoisoindole-1-carboxylic acid can replace proline in both sulfhydryl and non-sulfhydryl ACE inhibitors to give compounds equipotent to captopril and enalapril both in vitro and in vivo. Structure-activity relationships are discussed. Compound 11a (CI-907, indolapril) has advanced to clinical evaluation. PMID- 3035179 TI - High-affinity leukotriene receptor antagonists. Synthesis and pharmacological characterization of 2-hydroxy-3-[(2-carboxyethyl)thio]-3-[2-(8 phenyloctyl)phenyl] propanoic acid. PMID- 3035181 TI - Transmission and in vitro excretion of bluetongue virus serotype 1 by inoculated Culicoides brevitarsis (Diptera: Ceratopogonidae). PMID- 3035182 TI - Prevalence of enlarged salivary glands in Glossina palpalis, G. pallicera, and G. nigrofusca (Diptera: Glossinidae) from the Vavoua area, Ivory Coast. PMID- 3035183 TI - Localisation of Y chromosome sequences in normal and 'XX' males. AB - Three unique sequences derived from the Y chromosome have been mapped within the human genome. A Y specific sequence DYS20 is localised to Yq11.2. DXYS25 and DXYS27 are both X-Y homologous sequences which map to the Y short arm and to Xq21. DXYS25 maps more distally than DXYS27, on the Y short arm and on the X long arm. Y specific restriction fragments for these two sequences are shown to be present in the genome of two XX males, and an aberrant signal for DXYS25 is demonstrated at the tip of an X chromosome short arm in one XX male by in situ hybridisation. The implications of these findings for the location of the testis determining factor are discussed. PMID- 3035185 TI - The oxidase test in yeasts of medical importance. AB - The results of an oxidase test were positive with species of Cryptococcus and eight species of Candida, but negative with Candida krusei and species of Saccharomyces and Torulopsis, grown on Columbia Agar base and similar media, including Brain Heart Infusion Agar and Mueller Hinton Agar. When grown on Sabouraud Dextrose Agar, all the yeast genera and species were oxidase negative. The test is useful in delineating the genus Candida from Saccharomyces and Torulopsis. PMID- 3035184 TI - Cleft lip and palate, pili torti, malformed ears, partial syndactyly of fingers and toes, and mental retardation: a new syndrome? AB - Two sibs with a syndrome including cleft lip and palate, sparse scalp hair, malformed protruding ears, and partial syndactyly of the fingers and toes are reported. The older child also has mental retardation and pili torti. This syndrome is most probably inherited as an autosomal recessive disorder. PMID- 3035187 TI - B virus, Herpesvirus simiae: historical perspective. AB - Between 1932 and 1972, 24 known infections of man by B virus caused 23 cases of encephalitis and 18 fatalities. The virus has been isolated from dermal lesions and neural ganglia from macaque monkeys. Serological evidence of infection is complicated by close antigenic relationships between B virus, Herpesvirus simplex, and SA8. Hyperimmune globulin produced from monkey, horse, and rabbit sera has not proved highly effective. Formalin-inactivated vaccine appears safe and antigenic in man but has not been licensed. Half of all human subjects have neutralizing B virus antibody related to their H. simplex titer. More stable animal populations and the improved use of protective apparel have reduced, but not eliminated, the risk of B virus to man. PMID- 3035186 TI - Biosafety in acquired immunodeficiency syndrome (AIDS) studies using nonhuman primates. AB - The safety recommendations for studies on acquired immunodeficiency syndrome (AIDS) using nonhuman primates are based on knowledge about the epidemiology of the disease in humans, characteristics of the virus, and standard methods for handling nonhuman primates in the laboratory. Appropriate procedures avoid exposure to potentially infectious materials by skin puncture or to mucous membranes by using appropriate disinfecting agents, physical containment, protective clothing, and animal handling techniques. PMID- 3035189 TI - Mental outcome in West syndrome: prognostic value of some clinical factors. AB - Fifty-eight cases of West syndrome (eight idiopathic, 50 secondary) have been followed-up for a mean period of 5 years 5 months. The relationship has been studied between intellectual status at the final follow-up and the following variables: etiology, age of onset, neurological examination and developmental assessment at the onset, CT scan findings, short-term effect of ACTH treatment, and age of acquiring four developmental milestones (sitting unsupported, walking unsupported, uttering the first specific word and using a sentence of two words). Developmental and/or neurological abnormalities before the onset of the spasms, symptomatic etiology and abnormal CT findings are associated with a low IQ at the final follow-up. The age for sitting unsupported and for walking unsupported has proved to be not predictive of intellectual outcome, but the speech development data have shown a highly significant relationship with intellectual development. Early identification of the children with mental retardation has turned out to be superior to prediction of normal mental outcome. PMID- 3035190 TI - Phage T1-mediated transduction of a plasmid containing the T1 pac site. AB - The T1 pac site has been cloned into a plasmid vector. This recombinant plasmid was tested for T1-mediated transduction efficiency in comparison with a plasmid containing the phage lambda T1-pac-like site esp-lambda, plasmids containing T1 sequences other than the pac site, and plasmids containing neither T1 sequences nor known pac sites. The data obtained indicate that there are at least two distinct mechanisms of T1-mediated plasmid transduction. One requires the presence of any T1 sequence on the plasmid and probably takes place via cointegrate formation with the homologous region of an infecting T1 genome. The other is specifically dependent on the presence of a pac site on the plasmid. Plasmids are packaged as head-to-tail multimers that have one heterogeneous molecular end and the other terminated at pac, and the direction of packaging with respect to the pac site is the same for plasmids as for T1. Possible roles of pac in plasmid packaging and their implications with regard to the packaging of phage DNA are discussed. PMID- 3035191 TI - Complete sequence-specific 1H nuclear magnetic resonance assignments for the alpha-amylase polypeptide inhibitor tendamistat from Streptomyces tendae. AB - The 1H nuclear magnetic resonance (n.m.r.) spectrum of the alpha-amylase inhibitor Tendamistat was completely assigned with the use of phase-sensitive homonuclear two-dimensional n.m.r. The assignments include the non-labile protons of the 74 amino acid residues as well as the labile protons which exchange sufficiently slowly to be observed in H2O solution. The proton chemical shifts are listed at 50 degrees C and pH 3.2, which coincides with the conditions used for the determination of the three-dimensional structure of Tendamistat. PMID- 3035188 TI - The nature of the neutral Na+-Cl(-)-coupled entry at the apical membrane of rabbit gallbladder epithelium: I. Na+/H+, Cl-/HCO3- double exchange and Na+-Cl- symport. AB - Cl- influx at the luminal border of the epithelium of rabbit gallbladder was measured by 45-sec exposures to 36Cl- and 3H-sucrose (as extracellular marker). Its paracellular component was evaluated by the use of 25 mM SCN- which immediately and completely inhibits Cl- entry into the cell. Cellular influx was equal to 16.7 mu eq cm-2 hr-1 and decreased to 8.5 mu eq cm-2 hr-1 upon removal of HCO3- from the bathing media and by bubbling 100% O2 for 45 min. When HCO3- was present, cellular influx was again about halved by the action of 10(-4) M acetazolamide, 10(-5) to 10(-4) M furosemide, 10(-5) to 10(-4) M 4-acetamido-4' isothiocyanostilbene-2,2'-disulfonate (SITS), 10(-3) M amiloride. The effects of furosemide and SITS were tested at different concentrations of the inhibitor and with different exposure times: they were maximal at the concentrations reported above and nonadditive. In turn, the effects of amiloride and SITS were not additive. Acetazolamide reached its maximal action after an exposure of about 2 min. When exogenous HCO3- was absent, the residual cellular influx was insensitive to acetazolamide, furosemide and SITS. When exogenous HCO3- was present in the salines, Na+ removal from the mucosal side caused a slow decline of cellular Cl- influx; conversely, it immediately abolished cellular Cl- influx in the absence of HCO3-. In conclusion, about 50% of cellular influx is sensitive to HCO3-, inhibitable by SCN-, acetazolamide, furosemide, SITS and amiloride and furthermore slowly dependent on Na+. The residual cellular influx is insensitive to bicarbonate, inhibitable by SCN-, resistant to acetazolamide, furosemide, SITS and amiloride, and immediately dependent on Na+. Thus, about 50% of apical membrane NaCl influx appears to result from a Na+/H+ and Cl-/HCO3- exchange, whereas the residual influx seems to be due to Na+-Cl- cotransport on a single carrier. Whether both components are simultaneously present or the latter represents a cellular homeostatic counter-reaction to the inhibition of the former is not clear. PMID- 3035192 TI - Developmental regulation of cytosine methylation in the nuclear ribosomal RNA genes of Pisum sativum. AB - Prominent features of the cytosine methylation pattern of the Pisum sativum nuclear ribosomal RNA genes have been defined. Cytosine methylation within the C C-G-G sequence was studied using the restriction enzymes HpaII and MspI and gel blot hybridizations of the restriction digests. The extent to which particular features of the methylation pattern change during seedling development has also been determined. Total cellular DNA, purified from defined sections of pea seedlings grown under different lighting conditions, was analyzed with DNA hybridization probes derived from different portions of a cloned member of the nuclear rRNA gene family. By use of an indirect end-labeling technique, a map of 23 cleavable HpaII and/or MspI sites in genomic rDNA was constructed. The map covers about 90% of the rDNA repeat including the entire non-transcribed spacer region and most of the rRNA coding sequences. One notable feature of the map is that the most prominent HpaII site, located about 800 base-pairs upstream from the 5' end of the mature 18 S rRNA, is cleaved only in one of the two most abundant rDNA length variants (the short variant). With a gel blot assay specific for cleavage at this site, we estimated the HpaII sensitivity of DNA preparations from several stages of pea seedling development. We find that, while methylation is generally low in young seedlings, DNA obtained from the apical buds of pea seedlings is highly methylated. Further, the methylation level of rDNA within the pea bud decreases as the buds are allowed to develop under continuous white light. Our data, taken together with published studies on pea seedling development, indicate that cytosine methylation levels may be related to the regulated expression of the nuclear rRNA genes in pea. PMID- 3035193 TI - Sequences that adopt non-B-DNA conformation in form V DNA as probed by enzymic methylation. AB - pBR322 form V DNA is a highly torsionally strained molecule with a linking number of zero. We have used sequence-specific DNA methylases as probes for B-DNA in this molecule, exploiting the inability of methylases to methylate single stranded DNA and Z-DNA, both of which are known to occur in form V DNA. Some sequences in form V DNA were shown to be totally in the B-form, others were totally in an altered, unmethylatable conformation, while still other sites appeared to exist partly in altered and partly in normal B-conformation. Some potential Z-forming sequences (alternating pyrimidine/purine) of less than seven base-pairs were not in the Z conformation in form V DNA, whereas others did adopt an altered structure, indicating a modulating influence of flanking sequences. Furthermore, regions of imperfect alternating pyrimidine/purine structure were sometimes capable of adopting an altered structure. In addition, some regions of altered structure had no apparent Z-forming sequences, nor were they in polypurine stretches, which have also been proposed to form left-handed DNA. These non-B-DNA conformations may represent novel left-handed helical structures or sequences that become single stranded under torsional strain. Long regions of either altered (unmethylatable) DNA or B-DNA were not always observed. In fact, one region showed three transitions between B-like DNA and altered structure within 26 base-pairs. PMID- 3035194 TI - Structure of a gene for rat calmodulin. AB - The structural organization of the entire rat calmodulin gene was determined by cloning and sequencing overlapping genomic and cDNA clones from rat genomic and brain cDNA libraries. The intron/exon organization was determined by direct comparison of these sequences. Rat calmodulin gene is 9000 bases long and consisted of six exons interrupted by introns of variable sizes. The first intron separates the initiation codon (ATG) from the coding region of the protein. Three out of four intron/exon junctions in the coding region reside in the middle of calcium binding subdomains and do not correlate with the quarterly divided intramolecular homology of the protein. Their positions exactly coincide with those of the corrected version of chicken calmodulin gene. The rat calmodulin gene harbors a stretch of sequences homologous to a rat middle repetitive "identifier sequence" in the middle of the third intron. Analysis of the immediate 5' upstream region detected a TATA box (TATATATAT) and three C-G boxes (CCGCCC) but not a CAT box (CCAAT). A conserved sequence (GCGCCGCGYCYYGGGGGC) was found at -125 for rat and at -204 for chicken calmodulin genes. PMID- 3035195 TI - Mapping of sequence-specific chromatin proteins by a novel method: topoisomerase I on Tetrahymena ribosomal chromatin. AB - DNA derived from the 5' spacers of the rRNA genes from Tetrahymena has unusual electrophoretic properties. These properties made it possible to devise a simple electrophoretic procedure for isolating specific rDNA spacer fragments from preparations of total nuclear DNA, enabling us to study DNA modifications at the level of unfractionated nuclei. We have employed the method to study the distribution of topoisomerase I binding sites on the r-chromatin (ribosomal chromatin) of Tetrahymena at the DNA sequence level. The presence of topoisomerase I in situ was detected by its ability to introduce single-strand cleavages into DNA. The positions of the cleavages were determined on DNA sequencing gels after isolation of the fragments. Topoisomerase I binding in r chromatin is sequence specific and cleavage is confined to a 16 base-pair conserved sequence element previously determined to be a high-affinity binding site for topoisomerase I in vitro. The high degree of sequence specificity may be of important functional significance, as we find a similar sequence specificity with enzymes isolated from five evolutionarily distant species, indicating that preference for the 16 base-pair element is an intrinsic property of eukaryotic type I topoisomerases. PMID- 3035196 TI - Structure of the DNA gyrase-DNA complex as revealed by transient electric dichroism. AB - We have analyzed the structure of complexes between DNA gyrase and four defined DNA fragments by electric dichroism. Both the extrapolated dichroism and relaxation time of these complexes suggest that a single turn of DNA is wrapped around the enzyme with the entry and exit points located close together. The average angle between the DNA tails emerging from the particle is about 120 degrees. This structure is consistent with that seen by electron microscopy. Addition of ATP or the non-hydrolyzable ATP analog 5'-adenylyl-beta, gamma imidodiphosphate results in a structural change of the complex, consistent with the DNA tails now being wrapped around the protein. The significance of these observations with respect to the mechanism of DNA supercoiling by DNA gyrase is discussed. PMID- 3035197 TI - Response of the flagellar rotary motor to abrupt changes in extracellular pH. AB - Cells of a motile Streptococcus were starved, tethered to a quartz coverslip, energized with a potassium diffusion potential, and exposed to sudden decrements in external pH generated by flash photolysis of 2-hydroxyphenyl-1-(2-nitro)phenyl phosphate. The rotation rate of the cells increased following the flash but only after a brief time lag. Lags of the order of 0.1 second were observed in a dilute buffer (0.05 mM), confirming results obtained earlier. These lags were longer when the buffer was prepared in D2O. However, lags as short as 0.01 second were found in more concentrated buffers (1 and 3 mM). In this case, there was no deuterium solvent isotope effect. These differences arise from the extra time required for diffusion of protons from a dilute medium into the cell wall, which has a large buffering capacity. The short lags observed in concentrated media could be inherent to the flagellar motor, but the possibility that they are due to buffered diffusion through the cell wall or to elastic filtering by the tether has not been ruled out. PMID- 3035198 TI - Role of radiation therapy in small cell lung cancer: a bio-clinico-pathological review and perspective. AB - The role of radiotherapy in small cell carcinoma of the lung is unsettled; however, the radiosensitivity of this neoplasm is unquestioned. The ability of radiotherapy to cure or improve patients with this disease is still undergoing study. A review of this challenging subject is presented. PMID- 3035199 TI - Effect of micromolar concentrations of delta-9-tetrahydrocannabinol on herpes simplex virus type 2 replication in vitro. AB - The effect of micromolar concentrations of delta-9-tetrahydrocannabinol (delta-9 THC) on the in vitro replication and biosynthesis of herpes simplex virus type 2 (HSV2) was determined. A 100-fold increase in extracellular virus was recorded for infected Vero cells pretreated with 10(-6) M or 10(-5) M drug when compared to infected vehicle-treated controls. However, no significant differences were observed in the production of total infectious virus for any of the vehicle or drug-treated cultures. Immunofluorescence of virus-infected cells revealed that delta-9-THC did not alter the intracellular compartmentalization of virus specified proteins. Analytical sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography of isotopically labeled, cell-associated virus-specified proteins revealed that delta-9-THC had no major effect on the production of early nonstructural proteins but decreased the synthesis of late structural proteins. Scanning electron microscopy and light microscopy revealed blebs on the surface and macrovacuoles in the cytoplasm of both infected and uninfected cells treated with drug. These results suggest that delta-9-THC at micromolar concentrations selectively targets the host cell with the consequence of perturbation of cellular membranes. The alteration of cellular membranes may account for the enhanced virus release and for the decreased expression of virus specified, cell-associated late structural proteins. PMID- 3035200 TI - Mobile and immobile hydroxyapatite integration and resorption and its influence on bone. PMID- 3035201 TI - Receptors for nerve growth factor on rat spleen mononuclear cells. AB - Considerable evidence is mounting to support the concept of a modulatory role for the brain and neuroendocrine system on the immune response. This neuroimmunomodulation occurs in part through the interaction of specific neurosubstances with receptors on lymphocytes and monocytes. Nerve growth factor (NGF) is a neuronotrophic factor necessary for the development and maintenance of sympathetic and embryonic sensory neurons. This trophic effect is initiated through binding of NGF at specific cell surface receptor sites on NGF-responsive cells. Several recent studies suggest that NGF may interact with cells of the immune system and may play a role in the regulation of some immunologic reactions. In this study we report on the presence of specific receptors for NGF on the surface of mononuclear cells from rat spleens. The NGF-binding sites are of the low-affinity type with Kd's in the 10(-9) M range. These receptors migrate on SDS-PAGE as two molecular species of approximately 190 and 125 kilodaltons. Our findings of receptors for NGF on lymphocytes and accessory cells support other evidence that NGF may influence immunoreactivity in vivo. PMID- 3035204 TI - Meso-2,3-dimercaptosuccinic acid in the diagnosis and treatment of lead poisoning. AB - Lead poisoning remains one of the hazards of industrialized civilization. CaNa2 EDTA and dimercaprol, the usual therapeutic measures, have many side effects and can be given by parenteral route alone. The authors present a case of chronic lead poisoning caused by ingestion of contaminated flour ground in a primitive flour mill. The diagnosis was confirmed by the CaNa2 EDTA provocative test. Dimercaptosuccinic acid (DMSA) was given orally as a further provocation and resulted in an 11-fold increase in urinary lead excretion. A 5-day course of treatment with DMSA was instituted, during which symptoms abated, urinary lead excretion increased and the blood lead level decreased. No side effects were noticed. There has been no relapse over several months of follow-up. The authors conclude that the oral use of DMSA is effective, safe and convenient both as a provocative test in establishing the diagnosis of lead poisoning and as a therapeutic tool. PMID- 3035203 TI - Dissimilarities between benzodiazepine-binding sites and adenosine uptake sites in astrocytes and neurons in primary cultures. AB - The question whether the benzodiazepine receptor site in astrocytes or in neurons might be identical to the adenosine uptake site was studied by determining pharmacological profiles, inhibition types, and the effects of benzodiazepine antagonists in primary cultures of either astrocytes or neurons. Fourteen different benzodiazepines and five different adenosine uptake inhibitors displaced [3H] diazepam and inhibited adenosine uptake in both astrocytes and neurons. However, the rank orders (determined as IC50 values) with which these two parameters were affected were profoundly different, indicating dissimilarities between these two sites. For several of the compounds a difference in inhibition type (competitive vs. noncompetitive) was observed between the benzodiazepine-binding site and the adenosine uptake site in astrocytes and/or neurons, which further corroborated the conclusion of a difference between the benzodiazepine-binding site and the adenosine uptake site. Finally, the neuronal benzodiazepine antagonists RO 15-1788 and CGS-8216 and the astrocytic benzodiazepine antagonist PK 11195, which reverse the action of benzodiazepines, were not able to reverse inhibition of adenosine uptake by diazepam but exerted an inhibitory effect of their own. PMID- 3035202 TI - Transient increase in intracellular concentration of adenosine 3':5'-cyclic monophosphate results in morphological and biochemical differentiation of C6 glioma cells in culture. AB - Incubation of C6-BU1 glioma cells in the presence of isoproterenol and Ro20-1724- a potent cAMP phosphodiesterase inhibitor--results in a transient increase in intracellular cAMP levels, followed by a rapid efflux of cyclic AMP from the cells into the media. Two distinct types of morphological changes could be seen: rounded cell bodies with multipolar processes and beadings after 30 minutes of incubation--this period coincides with a 70-80-fold increase in intracellular cAMP levels, and elongated cell bodies with extended bipolar processes after 24 48 hours. By this time the intracellular cAMP concentration dropped to a low level, which was only three- to four fold higher than that in control. The transient increase in intracellular cAMP concentration results in retardation of cell growth, diminished uptake of 3H-2-deoxyglucose, and abolition of enhanced synthesis of cyclic AMP by concanavalin A. PMID- 3035205 TI - Clearance of ethylene glycol by kidneys and hemodialysis. AB - A patient with acute ethylene glycol poisoning was treated with ethanol administration and hemodialysis, and his renal function remained consistently normal. Serial measurements of serum and urine levels of urea, creatinine, ethylene glycol, and ethanol were performed to compare the relative contributions of the hemodialyzer and the patient's kidneys in clearing ethylene glycol from the blood. Simultaneous measurements of the serum-osmolal gap (corrected for ethanol) and anion gap were correlated with these data. Mean renal clearance of ethylene glycol was 27.5 +/- 4.1 ml/min, with a fractional ethylene glycol excretion of 19.8 +/- 1.5%. This was lower than the mean urea clearance of 89.4 +/- 11.0 ml/min and fractional urea excretion of 66.0 +/- 7.8%. Hemodialyzer clearance of ethylene glycol was 156 ml/min. There was a nearly exact correlation between the serum ethylene glycol level and the corrected osmolal gap (r = 0.998, p less than 0.01). The calculated renal elimination half-life of ethylene glycol was 18 hr. We conclude that with a moderate diuresis, the normal human kidney contributes significantly to the removal of ethylene glycol from the blood. Corrected serum osmolal gap provides a nearly exact approximation of the serum ethylene glycol level and is a useful therapeutic guide. PMID- 3035207 TI - Cutaneous seeding of hepatocellular carcinoma after fine-needle aspiration biopsy. PMID- 3035206 TI - Cytomegalovirus antibodies in breast milk and sera of mother-infant pairs. PMID- 3035208 TI - The 160,000-Mr virion protein encoded at the right end of the herpesvirus saimiri genome is homologous to the 140,000-Mr membrane antigen encoded at the left end of the Epstein-Barr virus genome. AB - The sequence of 4.4 kilobase pairs (kbp) from the conventional right terminus of the A + T-rich light-DNA (L-DNA) sequences of the herpesvirus saimiri (HVS) genome contains a leftward-directed open reading frame (ORF) for a 1,299-residue protein. The molecular weight predicted for the protein (143,000) is in good agreement with the estimates of 150,000 to 160,000 for the major nonglycosylated polypeptide of the virion tegument (the 160K polypeptide), previously shown to be encoded by this region of the genome. The first initiation codon of the ORF is only 250 nucleotides from the junction of the L-DNA component with the G + C-rich terminal reiterations (i.e., heavy or H-DNA) of the genome. An unusually A + T rich sequence (43 of 45 nucleotides are A or T, relative to a mean composition of 40% G + C for the ORF) occurs some 75 bp 5' to this initiation codon, and the first adenylation signal (AATAAA) on this DNA strand occurs 18 bp 3' to the termination codon. The amino acid sequence predicted for the 160K protein of HVS is homologous over most of its length to the 1,318-residue protein encoded by the leftmost major ORF of the G + C-rich genome of Epstein-Barr virus (BNRF1, the 140K nonglycosylated membrane antigen). No homology to either of these proteins is evident among the products predicted from the complete sequence of the alpha herpesvirus varicella-zoster virus. Thus gamma herpesviruses with coding sequences which differ in mean nucleotide composition by some 20% G + C have homologous proteins encoded at similar positions with respect to genome termini, with the right end of HVS being homologous to the left end of Epstein-Barr virus. PMID- 3035209 TI - Oligomerization and origin DNA-binding activity of simian virus 40 large T antigen. AB - Simian virus 40 (SV40) large tumor antigen (T antigen) exists in multiple molecular forms, some of which are separable by zone velocity sedimentation of soluble extracts from infected monkey cells. Three subclasses of this antigen from SV40-infected monkey cells have been separated and characterized: the 5S, 7S, and 14S forms. Newly synthesized T antigen occurs primarily in the 5S form. Chemical cross-linking provided evidence that the 14S form is primarily a tetramer, whereas the 5S and 7S forms could not be cross-linked into oligomers. The DNA-binding properties of each subclass were investigated after immunopurification. The affinities of the three forms for SV40 DNA and for a synthetic 19-base-pair sequence from binding site I are very similar (equilibrium dissociation constant [KD], 0.3 to 0.4 nM). The specific activity of DNA binding was greatest for the 5S and 7S subclasses and least for the 14S subclass. Moreover, the specific activity of the 5S and 7S subclasses increased sharply at about 40 h after infection, whereas the activity of the 14S subclass was maintained at a constant low level throughout infection. A model relating oligomerization and DNA binding of T antigen in infected cells is presented. PMID- 3035211 TI - Posttranslational processing of the Epstein-Barr virus-encoded p63/LMP protein. AB - In this paper we describe the posttranslational processing of the p63/LMP (latent membrane protein) encoded by Epstein-Barr virus in transformed B cells. Specifically, we show that after synthesis, free LMP disappeared with a half-life of about 0.5 h. This was caused by the association of LMP with an insoluble complex. All detectable LMP in the plasma membrane was insoluble. This interaction was resistant to nondenaturing detergents but readily dissociated with 8 M urea or by boiling in 0.5% sodium dodecyl sulfate, suggesting that LMP may be associated with cytoskeletal elements. Most of the Nonidet P-40-insoluble LMP was phosphorylated (ppLMP) primarily on serine but also on threonine residues. No phosphotyrosine was detected. Furthermore, greater than 90% of the ppLMP resided in the Nonidet P-40-insoluble fraction, suggesting a strong correlation between complexing and phosphorylation. Additionally, ppLMP was found to be associated with a 53,000-molecular-weight phosphoprotein (pp53) of unknown origin. Finally, LMP turned over extremely rapidly, with a half-life of about 2 h. Taken together, these properties suggest that although LMP falls broadly within the category of phosphorylated, cytoskeleton-associated oncoproteins, it is nevertheless clearly different from any previously described member of this family. PMID- 3035210 TI - B-lymphoma induction by reticuloendotheliosis virus: characterization of a mutated chicken syncytial virus provirus involved in c-myc activation. AB - Nondefective reticuloendotheliosis virus induces chicken bursal lymphoma in a manner similar to that of avian leukosis virus. The provirus integrates in the c myc locus and uses a promoter insertion mechanism to activate c-myc expression. We cloned a provirus involved in c-myc activation from a B lymphoma. Detailed structural characterization of this clone, including sequence determination, revealed proviral insertion at 512 base pairs preceding the second c-myc exon. The provirus has a deletion of 80% of the viral genes but retains two intact long terminal repeats (LTRs). A segment of the viral env sequence is present in an inverted orientation. Elevated expression of c-myc, apparently directed by the 3' LTR, was detected. However, despite the presence of an intact 5' LTR, no viral transcripts were detected. Thus, the internal proviral rearrangement can affect 5' LTR transcription or stability of the message or both. This finding is in consonance with the view that proviral deletion plays an important role in the induction of bursal lymphomas by nonacute retroviruses. PMID- 3035214 TI - Bovine papillomavirus transcriptional regulation: localization of the E2 responsive elements of the long control region. AB - The long control region (LCR) of the bovine papillomavirus type 1 genome can function as a conditional transcriptional enhancer which can be specifically trans-activated by the viral E2 gene product. To precisely map the target(s) of this trans-activation, BAL 31 exonuclease was used to generate two overlapping series of deleted DNA segments through the LCR. These fragments were assayed for their ability to activate transcription from the enhancer-deleted simian virus 40 early promoter of pA10CAT in the presence or absence of the viral E2 gene product. Two different E2-responsive elements were localized within the LCR. The major target for E2 trans-activation (E2-responsive element 1) was mapped to a 196-base-pair fragment between nucleotides 7611 and 7806, just upstream from promoters P7940 and P89. Further deletions which destroyed or impaired enhancer function revealed that the ACCN6GGT sequence motifs at each end of E2-responsive element 1 are critical components of this element. Primer extension analysis of RNA extracted from acute transfections with plasmids containing the bovine papillomavirus type 1 LCR driving the CAT gene revealed that each of the P7940 and P89 promoters is responsive to E2 trans-activation. PMID- 3035212 TI - Biologic and molecular characterization of two newly isolated ras-containing murine leukemia viruses. AB - A murine sarcoma virus (MSV) was recovered from an (NFS X NS.C58v-1) F1 mouse which developed splenic sarcoma and erythroleukemia 6 months after inoculation with a mink cell focus-inducing murine leukemia virus (MuLV) isolated from an NFS mouse infected with a wild mouse ecotropic MuLV. The MSV, designated NS.C58 MSV 1, induced foci of transformation in mouse and rat fibroblasts, and inoculation of mice of various strains 2 weeks of age or younger resulted in erythroleukemia and sarcomatous lesions in spleen, lymph node, and brain. The MSV provirus was molecularly cloned from a genomic library prepared from transformed non-producer rat cells. The 8.8-kilobase proviral DNA contained a 1.0-kilobase p21 ras coding segment which replaced most of the gp70-encoding portion of an MuLV, most likely the endogenous C58v-1 ecotropic virus. The ras oncogene is closely related to v Ha-ras by hybridization, expression of p21 protein, and nucleotide sequence. It is nearly identical in sequence to v-bas, the only previously described transduced, activated mouse c-ras. At position 12 in the p21 coding region, arginine is substituted for the naturally occurring glycine present in c-ras. A second MSV isolate is described which is similar to NS.C58 MSV-1 except for a 100 to 200-base-pair deletion in the noncoding region of the ras-containing insert. PMID- 3035213 TI - Regulation by recombinant interleukin-2 of protective immunity against recurrent herpes simplex virus type 2 genital infection in guinea pigs. AB - The goal of our study was to determine whether recombinant interleukin-2 (rIL-2) could modify the recurrence pattern of chronic herpes simplex virus type 2 (HSV 2) genital infection in guinea pigs. Animals that developed symptomatic acute HSV 2 infection were distributed at 14 days after viral inoculation into several treatment groups, which were similar with respect to the severity of acute disease. Three rIL-2 dosages administered for 4 weeks in daily subcutaneous injections were tested in this study: 5 X 10(3), 5 X 10(4), and 2.5 X 10(5) U. Daily observations of the animals showed a significant decrease of the incidence of new recurrent lesions with the use of 5 X 10(4) U of rIL-2 (rate of recurrence, 0.08, compared with 0.21 in untreated controls), whereas the other rIL-2 regimens did not affect the overall rate of recurrence. Weekly analysis of recurrences showed that treatment with 5 X 10(4) U of rIL-2 was effective only during the first 3 weeks of use and that 2.5 X 10(5) U of rIL-2 markedly decreased the rate of recurrence in the first week of treatment but not in subsequent weeks. The loss of clinical protection in both groups coincided with the production of neutralizing antibodies to rIL-2. The immune mechanisms possibly involved in the protective effect of rIL-2 in chronic HSV-2 disease were further investigated. Production of gamma interferon correlated well with clinical protection, and circulating levels dropped at the time when neutralizing antibodies to rIL-2 developed. Nonspecific cytotoxicity represented by natural killer cell and lymphokine-activated killer cell activities was also increased in the treated guinea pigs. Antibody titers and lymphocyte proliferation to herpes simplex antigen were similar in rIL-2 and placebo recipients. Finally, we found that the rIL-2-induced immune stimulation was as protective against recurrent HSV 2 disease in guinea pigs as the viral suppression achieved with acyclovir. However, the biological activity of both drugs was not additive when they were coadministered. PMID- 3035215 TI - The T-antigen-binding domain of the simian virus 40 core origin of replication. AB - The simian virus 40 origin of replication contains a 27-base-pair palindrome with the sequence 5'-CA-GAGGC-C-GAGGC-G-GCCTC-G-GCCTC-TG-3'. The four 5'-GAGGC-3'/5' GCCTC-3' pentanucleotides are known contact sites for simian virus 40 T-antigen binding in vitro. We used oligonucleotide-directed cassette mutagenesis to identify features of this palindrome that are important for the initiation of DNA replication in vivo. Each base pair of a pentanucleotide is crucial for DNA replication. In contrast, sequences adjacent to pentanucleotides have little or no effect on replication. Thus, the pentanucleotide is the basic functional unit, not only for T-antigen binding but also for DNA replication. All four pentanucleotides are indispensable in the initiation process. The spacing of pentanucleotides is crucial because duplication of the single base pair between binding sites has a far greater effect on replication than does substitution of the same base pair. Inversion of any pentanucleotide blocks DNA synthesis. Thus, the pentanucleotide is not a functionally symmetrical unit. We propose that each pentanucleotide positions a monomer of T antigen at the proper distance, rotation, and orientation relative to other T-antigen monomers and to other origin domains and that such positioning leads to subsequent events in replication. PMID- 3035216 TI - Site-directed mutagenesis of proteinase 3C results in a poliovirus deficient in synthesis of viral RNA polymerase. AB - We used a synthetic double-stranded oligonucleotide to introduce amino acid substitutions into the proteinase 3C region of a poliovirus type 1 cDNA clone. The six different mutant viruses recovered exhibited a small-plaque phenotype when assayed on HeLa cells. Further investigation revealed that all the mutations (with the exception of one) yielded P3 region proteins that displayed altered mobility in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A conservative Val----Ala change at amino acid 54 of the proteinase resulted in a virus that was deficient in the production of the mature viral RNA polymerase 3D. Although this mutant achieved less than one-half of the wild-type levels of RNA synthesis during the course of infection, it still grew to nearly wild-type titers. PMID- 3035217 TI - Trigeminal ganglion infection by thymidine kinase-negative mutants of herpes simplex virus after in vivo complementation. AB - Infection of trigeminal ganglion by herpes simplex virus (HSV) thymidine kinase negative (TK-) mutants was investigated in mixed infection studies in mice. Mice were corneally inoculated with TK- HSV alone or with mixtures of TK- HSV-TK+ HSV. When inoculated alone, an arabinosylthymine-selected HSV type 1 TK- mutant and a HSV type 2 TK- deletion mutant infected mouse ocular tissues but rarely infected ganglion tissues. However, both TK- mutants readily infected ganglion tissues when they were inoculated in mixtures with TK+ HSV. By means of mixed infection studies, it was demonstrated that TK- HSV could readily establish acute and latent ganglion infections. It was thought that the frequent infection of trigeminal ganglion tissue by both TK- mutants after mixed TK(-)-TK+ HSV infection was the result of in vivo complementation. After mixed TK(-)-TK+ HSV infection and subsequent cultivation of ganglion explants in arabinosylthymine, results supported the conclusion that when TK- was present in ganglia it was in the same neurons that contained TK+ HSV. PMID- 3035218 TI - Identification of p40x-responsive regulatory sequences within the human T-cell leukemia virus type I long terminal repeat. AB - Distinct transcriptional regulatory sequences located within the upstream sequences required for p40x trans-activation of the human T-cell leukemia virus type I (HTLV-I) long terminal repeat (LTR) were chemically synthesized and cloned upstream of the basal HTLV-I LTR promoter. Plasmids containing a single 21-base pair (bp) repeat were weakly inducible by p40x. The level of trans-activation by p40x was increased when two (30-fold) or three (40-fold) 21-bp repeats were present in the upstream control region. In the mutant containing two 21-bp repeats, the upstream 21-bp repeat could be positioned in either the sense (30 fold) or the antisense (16-fold) orientation. Plasmids containing a 51-bp repeat element, which included a single 21-bp repeat, were induced to levels similar to that obtained with the 21-bp repeat sequence alone. Template DNAs containing a single copy of the HTLV-I sequences between -117 and -160 were stimulated approximately 10-fold by p40x when one copy of the 21-bp element was located downstream. PMID- 3035219 TI - Clonal dominance and progression in Abelson murine leukemia virus lymphomagenesis. AB - We examined the clonality of tumors induced by an acutely transforming retrovirus which carries a single oncogene. Contrary to our expectation, tumors induced by the Abelson murine leukemia virus (A-MuLV) showed one to four major proviral integration events. To further investigate the process by which clonality was established, we analyzed the number of cells infected and transformed by A-MuLV at various times after in vivo infection. At the midpoint of tumor latency (14 days postinfection), we found that infection of total bone marrow cells by A-MuLV was efficient and polyclonal. However, only a minority of these infected cells were transformed as assayed in cell culture, and clonal dominance had already been established in this transformed cell population. Examination of the in vitro growth properties of transformed cells recovered from preleukemic and leukemic mice indicated that preleukemic cells had lower cloning efficiencies than primary tumor cells. Our results suggest that the rate-limiting step in this system of lymphomagenesis is the initial transformation of bone marrow target cells and that these cells undergo subsequent changes in cloning ability during the course of the disease that lead to an autonomous neoplastic state. PMID- 3035220 TI - Effects of herpes simplex virus on mRNA stability. AB - Herpes simplex virus virions contain one or more functions which mediate shutoff of host protein synthesis, disaggregation of host polyribosomes, and degradation of host mRNA. We studied aspects of the host shutoff mechanism by using herpes simplex virus type 1 mutants deficient in virion-induced shutoff of host protein synthesis (G. S. Read and N. Frenkel, J. Virol. 46:498-512, 1983). Shutoff of host protein synthesis by the wild-type virus was associated with degradation of host mRNAs, including beta-actin, alpha-tubulin, and heat shock protein 70. In contrast, the virion host shutoff (vhs) mutants were deficient to various degrees in their ability to induce host mRNA degradation; the extent of mRNA degradation correlated well with the extent of inhibition of host protein synthesis. This finding suggests that inhibition of host protein synthesis and degradation of host mRNA were mediated by the same virion-associated function. Virion-induced degradation of host mRNA was not prevented by inhibitors of ribosome translocation, nor could it be augmented, for mutant vhs-1, by drugs which disaggregate polyribosomes. This suggests that mRNA in polyribosomes, as well as nonpolyribosomal mRNA, is susceptible to virion-induced degradation. Finally, the half-life of viral transcripts was also prolonged in cells infected with the vhs 1 mutant virus, suggesting that the vhs function indiscriminately decreased the half-lives of both host and viral mRNAs. The vhs function may thus play a dual role in virus infection. (i) It inhibits host gene expression, and (ii) it enables rapid transitions in the expression of viral genes which are sequentially transcribed as infection progresses. PMID- 3035221 TI - Identification of a novel herpes simplex virus type 1-induced glycoprotein which complexes with gE and binds immunoglobulin. AB - We detected a glycoprotein on the surface of cells infected with herpes simplex virus type 1 (HSV-1) which, in conjunction with gE, binds immunoglobulin G (IgG). The novel glycoprotein, which has an apparent molecular mass of 70 kilodaltons and was provisionally named g70, was first detected in extracts of HSV-1-infected cells labeled by lactoperoxidase-catalyzed iodination and precipitated with rabbit sera or IgG and protein A-Sepharose. In subsequent experiments, g70 and gE were coprecipitated from extracts of HSV-1-infected cells labeled with [35S]methionine, [35S]cysteine, or 14C-amino acids. We were unable to precipitate a polypeptide analogous to g70 or gE from extracts of HSV-2-infected cells with rabbit IgG and protein A-Sepharose. Partial proteolytic peptide analysis indicated that g70 is structurally distinct from gE and gI). In addition, g70 was electrophoretically distinct from the HSV-1 Us4 glycoprotein gG. HSV-1 gE, expressed in mouse cells transfected with the gE gene, was not precipitated with rabbit IgG, nor could these cells bind radiolabeled IgG, suggesting that gE alone cannot act as an IgG (Fc) receptor. This result, coupled with the findings that gE and g70 are coprecipitated with IgG and with an anti-gE monoclonal antibody, suggests that gE and g70 form a complex which binds IgG. The electrophoretic mobilities of g70 molecules induced by different strains of HSV-1 differed markedly, arguing that g70 is encoded by the virus and is not a cellular protein induced by virus infection. PMID- 3035222 TI - Amphotropic proviral envelope sequences are absent from the Mus germ line. AB - We derived an amphotropic murine leukemia virus (MuLV) type-specific probe for use in Southern blot hybridizations with cloned and genomic DNAs. A 133-base-pair RsaI-RsaI fragment from the 5' env region of the amphotropic viral isolate 4070A was subcloned into M13mp18 and radiolabeled in vitro. The probe detected the proviral DNAs in mink cells infected with seven different amphotropic MuLV isolates. The probe did not cross hybridize with the DNAs of molecular clones of ecotropic, mink cell focus-forming, or xenotropic MuLVs; nor did it anneal to the proviral DNAs of four xenotropic or six mink cell focus-forming viral isolates grown in mink cells. DNAs of 12 inbred laboratory mouse strains and more than 15 different wild mouse species and subspecies were examined for the presence of endogenous amphotropic env-related fragments. Amphotropic env-related sequences were found only in the DNAs of wild mice trapped in southern California in an area previously shown to harbor mice producing infectious amphotropic virus. Restriction enzyme analyses of DNAs from these mice showed that amphotropic sequences were not present as germ line copies but were the result of congenital or horizontal infection or both in this population. The DNAs of 11 various mammalian and avian species, including both natural predators of mice and squabs from the farms with virus-positive mice, lacked amphotropic envelope-related sequences. PMID- 3035223 TI - Polyomavirus tumor induction in mice: influences of viral coding and noncoding sequences on tumor profiles. AB - We determined the DNA sequences of the noncoding regions of two polyomavirus strains that differ profoundly in their abilities to induce tumors in mice. Differences between strains were found, both on the late side of the replication origin in the region containing known enhancer elements and on the early side of the origin, affecting the number and location of large-T-antigen-binding sites. By constructing and analyzing recombinant viruses between these high- and low tumor strains, we attempted to localize determinants which affect the frequency and histotype of tumors. Seven recombinants were constructed and propagated in vitro, and the tumor profile of each was established by inoculation into newborn C3H mice. Recombinants containing noncoding sequences from the high-tumor strain and coding sequences from the low-tumor strain behaved like the latter, inducing tumors at a low frequency and strictly of mesenchymal origin. Reciprocal recombinants with noncoding sequences of the low-tumor strain linked to structural determinants from the high-tumor strain induced several types of epithelial tumors typical of the high-tumor strain but at reduced frequency, in addition to mesenchymal tumors. A high frequency and full diversity of epithelial tumors required, in addition to structural regions from the high-tumor strain, noncoding sequences on the early side of the origin also present in this strain. A high-tumor profile thus resulted from the combined effects of structural and regulatory determinants in the high-tumor strain, with the former affecting primarily the tissue tropism and the latter affecting the frequency of tumors. No differential effects of the enhancer regions from the late side of the origin in the two virus strains were seen in this study. PMID- 3035224 TI - Cooperation of middle and small T antigens of polyomavirus in transformation of established fibroblast and epithelial-like cell lines. AB - We have reported recently that small T antigen of polyomavirus stimulates the growth of NIH 3T3 cells beyond their saturation density and induces weak anchorage-independent growth (T. Noda, M. Satake, T. Robins, and Y. Ito, J. Virol. 60:105-113, 1986). We examined whether small T antigen would cooperate with middle T antigen in the in vitro transformation of NIH 3T3 (fibroblasts) and NRK-52E (epitheliallike) cells. The small-T-antigen gene, when cotransfected with the middle-T-antigen gene, had no additional effect on the efficiency or size of dense foci formation induced by the middle-T-antigen gene on a monolayer of NIH 3T3 cells. However, the small-T-antigen gene dramatically increased the rate of growth of NIH 3T3 cells transformed by middle T antigen in semisolid medium. Introduction of the small-T-antigen gene into middle-T-antigen-transformed cells did not disturb the integrated middle-T gene, alter expression of the middle-T gene, or enhance middle-T-antigen-associated tyrosine protein kinase activity. For NRK-52E cells, the expression of middle T antigen alone resulted in small, slow-growing foci on a monolayer. These cells did not show anchorage-independent growth, despite the fact that middle-T-antigen-associated tyrosine protein kinase activity was clearly detected in these cells. NRK-52E cells expressing both middle and small T antigens formed faster growing foci on a monolayer than middle T-antigen-expressing cells did and grew in semisolid medium, even when the amounts of middle T antigen and its associated kinase activities were lower than those of middle-T-antigen-expressing cells. We conclude that small T antigen cooperates with middle T antigen in the in vitro transformation of established cell lines of fibroblast and epitheliallike cells, that it does not share the middle-T-antigen function even though they are structurally related, and that it has a significantly more important role in the transformation of NRK-52E cells than that of NIH 3T3 cells. PMID- 3035225 TI - Divergent transcription of early 35- and 94-kilodalton protein genes encoded by the HindIII K genome fragment of the baculovirus Autographa californica nuclear polyhedrosis virus. AB - The organization of viral genes within the 3.7-kilobase-pair HindIII-K/EcoRI-S region of the Autographa californica nuclear polyhedrosis virus genome (85 to 88 map units) was determined by using a combination of nucleotide sequencing, transcriptional mapping, and in vitro translation of hybrid selected RNA. Two nonoverlapping genes, extending in opposite directions and encoding polypeptides with molecular weights of 35,000 and 94,000 (35K and 94K polypeptides), were identified. Unspliced, messenger-active RNAs were transcribed from both genes early (2 h) after infection. Indicative of immediate-early genes, transcription of the divergent RNAs was unaffected by the protein synthesis inhibitor, cycloheximide. Late in infection, abundant RNAs were transcribed from promoters located at least 2.5 kilobase pairs upstream from the gene encoding the 35K polypeptide. These transcripts completely overlapped both the 35K and 94K polypeptide genes but apparently lacked protein-coding potential, suggesting that the transcripts may play a role in the suppression of early viral gene expression. PMID- 3035227 TI - Localization of Escherichia coli RNA polymerase-binding sites on bacteriophage S13 replicative form I DNA by protection of restriction enzyme cleavage sites. AB - Protection of restriction endonuclease cleavage sites by Escherichia coli RNA polymerase bound to the replicative form I of bacteriophage S13 DNA has been used to identify a number of regions of RNA polymerase binding. Digestion with HincII, AluI, HinfI, or HaeIII, under conditions optimized for "open" complex formation, revealed 12 regions of RNA polymerase binding. Based on differential salt sensitivities, five of the regions were classified as strong or tight binding sites. These were located before genes A (two sites), B, and D and at the 5' end of gene F. The seven regions which exhibited weaker binding were located at the 5' end of gene C (two sites), in the middle of gene D, just before and at the 3' end of gene F, at the 5' end of gene G, and in the middle of gene H. The sites before genes B and D coincide with sites previously identified as promoters in bacteriophage phi X174. One of the sites before gene A, that at nucleotides 5175 5211, represents a new putative promoter site in bacteriophage S13 and phi X174 located before the previously identified A gene promoter at nucleotides 10-45. PMID- 3035226 TI - Herpes simplex virus immediate-early promoters are responsive to virus and cell trans-acting factors. AB - The promoters for each of the immediate-early genes from herpes simplex virus type 1 were cloned and fused to a chloramphenicol acetyltransferase cassette. These chimeric genes were used as targets in a transient expression assay to determine how the immediate-early gene products ICP4 and ICP0 and the virion associated stimulatory protein Vmw65 affected their expression in HeLa and Vero cells. The basal level of expression from these cassettes differed significantly depending on the extent of 5'-flanking sequence and the cell line that served as host. The promoters from IE-4 and IE-0 behaved in a qualitatively similar fashion independent of the host cell. However, the promoter for ICP27 had a unique response pattern: in Vero cells it acted as an alpha gene promoter, whereas in HeLa cells its response was more like that of a beta gene promoter. The promoter sequences for ICP22 and ICP47 behaved as the IE-4 and IE-0 promoters did in HeLa cells, but their response to the effector molecules in Vero cells was unlike that of other alpha gene promoters we have studied. Evidence is also presented for a role for ICP27 in autoregulation. PMID- 3035228 TI - Genetic reassortants for identification of the genome segment coding for the bluetongue virus hemagglutinin. AB - Two bluetongue virus (BTV) serotypes isolated in Australia and two selected reassortants derived from cells coinfected with these viruses have been used to identify the gene coding for the virus hemagglutinin. The parent viruses had characteristic hemagglutination patterns: BTV type 20 agglutinated sheep erythrocytes only; and BTV type 21 agglutinated sheep, bovine, human, and goose erythrocytes. Analysis of the two virus clones that had reassorted in genes coding for the outer capsid polypeptides demonstrated that hemagglutination and hemagglutination inhibition are functions associated with the outer capsid protein (VP2), which is encoded by genome segment 2. PMID- 3035229 TI - Poliovirus snapback double-stranded RNA isolated from infected HeLa cells is deficient in poly(A). AB - A portion of poliovirus double-stranded RNA (25 to 50%) isolated from infected HeLa cells contains hairpin loops at one end of the duplex structure. These structures rapidly reformed double-stranded molecules after denaturation and appeared as molecules of up to two times genome length upon electrophoresis in denaturing agarose gels. A second form of poliovirus double-stranded RNA was readily denaturable into genome length strands. When the hairpin RNA was treated with S1 nuclease, subsequent denaturation resulted in formation of strands of up to genome length. Hairpin molecules contained very little, if any, poly(A) sequences, suggesting that the hairpin forms after nucleolytic removal of the 3' end of plus-strand templates. We conclude that the hairpin double-stranded RNA found in infected cells is likely generated by intracellular nicking and self priming and that it does not represent an intermediate in the process of RNA replication. PMID- 3035230 TI - Nerve growth factor deprivation results in the reactivation of latent herpes simplex virus in vitro. AB - Primary sympathetic neuronal cultures were maintained for up to 5 weeks after inoculation with herpes simplex virus (HSV) without evidence of viral infection. Upon deprivation of nerve growth factor, the cultures produced infectious HSV, indicating that the cultures harbored latent HSV. This study demonstrates a function of nerve growth factor in the maintenance of HSV latency. PMID- 3035231 TI - The A+T-rich sequence of the simian virus 40 origin is essential for replication and is involved in bending of the viral DNA. AB - The origin-promoter region of simian virus 40 contains a 17-base-pair sequence composed exclusively of adenine (A) and thymine (T). We constructed a linker replacement mutant in which this stretch of A's and T's was reduced to 11 base pairs. While not affecting the level of early gene transcription, this mutation reduced the accumulation of viral DNA in COS cells at least 10(4) fold. In addition, a restriction fragment containing the wild-type A + T-rich region migrated in nondenaturing polyacrylamide gels with an anomalous mobility characteristic of bent DNA; however, the corresponding fragment from the mutant migrated less anomalously. Therefore, bending of the DNA in this region may play a role in some step in viral DNA replication. PMID- 3035232 TI - Selective translation initiation on bicistronic simian virus 40 late mRNA. AB - We described previously a simian virus 40 (SV40) mutant, pSVAdL, that was defective in synthesis of the late viral protein VP1. This mutant, which contains a 100-base-pair fragment of adenovirus DNA encompassing the major late promoter inserted in the SV40 late promoter region (SV40 nucleotide 294), efficiently synthesizes agnoprotein, a protein encoded by the leader region of the same mRNA that encodes VP1. When the agnoprotein AUG initiation codon in pSVAdL was mutated to UUG, agnoprotein synthesis was abolished, and VP1 synthesis was elevated to wild-type levels. Because levels of late mRNA synthesis were not affected by this mutation, these results support a scanning model of translation initiation and suggest that internal translational reinitiation does not occur efficiently in this situation. PMID- 3035233 TI - Two phosphorylated subclasses of polyomavirus large T antigen that differ in their modes of association with the cell nucleus. AB - Two classes of polyomavirus large T antigen were distinguished, differing in their modes of association with the cell nucleus. A weakly associated class, the nucleoplasmic T antigen, representing 30 to 40% of the total, was solubilized when cells were lysed isotonic buffer at pH 7.2. A more tightly bound class retained in isolated nuclei, the retained T antigen, was extractable either at pH 9.0 or in 2 M NaCl. The retained T antigen contained an additional mole of phosphate, 4 mol of PO4 per mol of T antigen, compared with the nucleoplasmic T antigen (3 mol of PO4 per mol of T antigen). Limit digestion with staphylococcal V8 protease yielded equivalent amounts of five peptides ranging in size from 7.5 to 20 kilodaltons. Additional phosphorylation within a 12-kilodalton peptide accounted for most of the difference in phosphate content between retained and nucleoplasmic T-antigen classes. PMID- 3035235 TI - Value of percutaneous core needle biopsy in the differential diagnosis of renal transplant dysfunction. AB - The value of percutaneous core needle biopsy in the differentiation of rejection from other causes of renal allograft dysfunction, and its subsequent effect on patient management were assessed in 64 consecutive biopsies performed on 34 patients in whom the clinical diagnosis was was uncertain. A complete clinical, biochemical and radiographic assessment was made in each patient before biopsy. Only 1 biopsy (1.6 per cent) yielded tissue inadequate for evaluation, while another biopsy caused a renal artery pseudoaneurysm that ruptured and resulted in graft loss. In 27 of these 64 biopsies (42 per cent) the results differed from the pre-biopsy diagnosis and directly affected patient management, particularly the use of steroids. The remaining biopsy specimens were helpful to confirm uncertain clinical impressions, and allowed accurate counseling for patients and family. Biopsies were of special usefulness in separating acute rejection from complications, such as acute tubular necrosis, cytomegalovirus infections, recurrence of original disease, cyclosporin toxicity and acute superimposed-upon chronic rejection. Of 64 biopsies 22 (34.3 per cent) demonstrated the absence of rejection and 8 demonstrated chronic rejection (12.5 per cent), thereby averting the use of steroids in 46.8 per cent of the patients. All patients with evidence of severe small vessel disease and/or antibody-mediated rejection eventually lost the grafts, including 2 with cytomegalovirus glomerulopathy who also suffered such vascular changes. These data highlight the extreme usefulness of needle biopsy in the evaluation and management of renal allograft dysfunction. PMID- 3035234 TI - Methylation of specific cytosine residues enhances simian virus 40 T-antigen binding to origin region DNA. AB - Specific binding of simian virus 40 large T antigen to origin region DNA requires the interaction of T antigen with multiples of a consensus recognition pentanucleotide sequence (5'-G[T]-A[G]-G-G-C-3'). To assess the interaction of T antigen with cytosine residues in the recognition sequences, bacterial methylases were used to methylate simian virus 40 form I DNA in vitro at specific cytosine residues. Methylation of a subset of the cytosine residues in the pentanucleotide sequences resulted in enhanced binding of T antigen to origin region DNA. Enhanced binding to the methylated pentanucleotides indicates that the methyl groups introduced on this subset of pentanucleotide cytosine residues could not have sterically interfered with the interaction of T antigen with the recognition sequences. This lack of steric interference suggests that T antigen does not make close contact in the major groove with these particular cytosine residues during normal binding. PMID- 3035236 TI - Correlation of computerized tomographic changes and histological findings in 80 patients having radical retroperitoneal lymph node dissection after chemotherapy for testis cancer. AB - A total of 80 patients with stage B3 or B2/C germ cell testis tumors underwent computerized tomography before and after chemotherapy. The volume and computerized tomographic density of metastatic retroperitoneal tumor were measured on all scans. The patients then underwent full bilateral retroperitoneal lymphadenectomy. The change in volume and density of retroperitoneal disease was correlated with the histological type of the primary testis tumor and with the histological findings at retroperitoneal lymphadenectomy. In all 15 patients (100 per cent) without teratomatous elements in the original tumor and who had a greater than 90 per cent decrease in the volume of retroperitoneal masses as a response to systemic chemotherapy no teratoma or active cancer was found in the surgical specimen. In contrast, 7 of 9 patients (78 per cent) with teratomatous elements in the original specimen had either teratoma or carcinoma in the retroperitoneal lymphadenectomy specimens despite having a greater than 90 per cent decrease in tumor volume. This difference was significant (p less than 0.05). These data suggest that patients with no teratomatous elements in the original specimen and a greater than 90 per cent decrease in the volume of retroperitoneal masses in response to chemotherapy can be observed carefully for signs of recurrence rather than undergoing post-chemotherapy retroperitoneal lymphadenectomy. PMID- 3035237 TI - Pulmonary emboli as a complication of germ cell cancer treatment. AB - A total of 3 patients with germ cell cancer had pulmonary emboli while receiving cisplatin-containing chemotherapy. In addition to cisplatin, 1 patient was receiving etoposide plus doxorubicin, 1 vinblastine plus bleomycin and 1 etoposide plus bleomycin at the time of the vascular event. One patient died of cardiovascular collapse, while the other 2 presented with severe shortness of breath, hemoptysis and pleuritic chest pain. A review of vascular complications of cisplatin-containing chemotherapy is presented. Awareness and early recognition of pulmonary emboli in patients receiving these chemotherapeutic agents may minimize treatment-related morbidity and mortality. PMID- 3035238 TI - Development and characterization of a monoclonal antibody to human embryonal carcinoma. AB - A monoclonal anti-testicular carcinoma antibody was obtained via the somatic cell fusion technique by immunization of BALB/c mice with freshly prepared single cell suspension from a patient with testicular embryonal carcinoma with choriocarcinoma components. The hybridoma supernates were screened against the testicular carcinoma cells used in the immunization as well as normal mononuclear white blood cells isolated from the same patient. An antibody (5F9) was selected which bound to fresh tumor cells from two patients with embryonal testicular carcinoma and failed to bind to fresh tumor cells from 24 patients (2 seminoma, 2 melanoma, 3 neck, 2 esophageal, 1 ovarian, 3 colon, 1 prostate, 2 breast, 1 liposarcoma, 3 endometrial, 1 kidney, 1 adrenal, 1 larynx and 1 bladder tumors) or cell suspensions prepared from normal liver, lung, spleen, ovary, testes, kidney, red blood cells or white blood cells. The antibody was tested for its binding to several well established cancer cell lines, and was found to bind to the BeWo human choriocarcinoma and two human embryonal carcinoma cell lines. The antibody did not react with 22 other cell lines or with hCG. The antibody was labeled with 131I and injected into nude mice bearing BeWo tumors and evaluated for tumor localization by performing whole body scans with a gamma camera 5 days later. Six mice injected with the antibody showed positive tumor localization without the need for background subtraction while six mice injected with MOPC-21, a murine myeloma immunoglobulin, demonstrated much less tumor localization. Tissue distribution studies performed after scanning showed specific tumor localization (8:1 tumor: muscle) for the monoclonal antibody and no specific localization for MOPC-21. This antibody thus has selective reactivity with the surface of tumor cells from embryonal carcinoma (testicle) and choriocarcinoma both in vitro and in vivo. PMID- 3035239 TI - 1,25-Dihydroxyvitamin D3 receptors and their relationship to histological features in renal cell carcinoma. AB - The present studies were carried out in 11 human renal cell carcinomas to determine the presence of a receptor specific for an active form of vitamin D, 1,25-dihydroxyvitamin D3. Saturation and Scatchard analyses of the cytosol receptor for 1,25-dihydroxyvitamin D3 showed that nine tumors had a detectable level of the receptor (two fmol/mg. protein). The equilibrium dissociation constant of these receptors ranged between 46 and 380 pM and the binding capacity also ranged between 3.5 and 12.7 femtomol/mg. protein. Sucrose density gradient analysis of the specific binders revealed that the tumors had a receptor protein appearing as a single 3.6S peak. Two tumors which had only a trace of the receptor were high grade solid tumors consisting mainly of spindle or pleomorphic cells. Nine tumors possessing 1,25-dihydroxyvitamin D3 receptor consisted of clear and/or granular cells. Thus, the absence of the receptor was only accompanied by low differentiated sarcomatoid tumors with poor prognosis. However, so far, the amount of the receptor in the receptor-positive tumor did not relate to the other clinical and pathological features of the patients. PMID- 3035240 TI - Experimental infection and horizontal transmission of Modoc virus in deer mice (Peromyscus maniculatus). AB - Deer mice (Peromyscus maniculatus) were inoculated with a sublethal dose of a field strain of Modoc virus to determine patterns of viral persistence, shedding, and transmission. Blood, serum, urine, fecal, and oral swab samples were collected at selected intervals until 63 days postinoculation (PI) after which lung, liver, spleen, kidney, and salivary glands were explanted. Viral assays were conducted by intracranial inoculations of suckling mice and antibody titers were determined by the micro-complement-fixation test. Viremias lasted for up to 4 days PI. Antibody titers were present by day 8 PI, peaked at day 13-20 PI, and persisted until day 63 PI. There was no evidence of viral shedding in urine, fecal, or oral swab samples. Virus was detected in explanted lungs only. In a separate experiment, deer mice were inoculated with virus and lungs were removed from five mice per wk for 10 wk. Indirect fluorescent antibody (IFA) techniques were used to determine the location of virus in lung tissue and to examine fixed tissue for lesions. IFA showed virus in lung parenchymal cells beginning 42 days PI and persisting at least 70 days PI. No histopathologic changes were seen. Horizontal transmission of the virus was studied by placing uninoculated mice with inoculated mice for 42 days and determining if the test animals developed antibodies or had virus in their lungs. Fifty-percent of the uninoculated mice developed antibody. One of these animals had virus in its lungs. Therefore, Modoc virus may be transmitted by direct contact. PMID- 3035241 TI - Epizootic vesicular stomatitis in Colorado, 1982: some observations on the possible role of wildlife populations in an enzootic maintenance cycle. AB - Sera obtained from wild ungulates, carnivores, and rodents in Colorado were tested for neutralizing (N) antibody against vesicular stomatitis, New Jersey serotype (VSNJ), virus to determine their involvement in the 1982 Colorado VSNJ epizootic in domestic animals. Viremic and N antibody responses of two local rodent species to a 1982 Colorado isolate of VSNJ were determined in the laboratory. The rodents produced only weak viremias, but all developed N antibody. N antibody prevalences for VSNJ in sera from wild ungulates was sufficiently high to indicate their involvement during the epizootic. In addition, the demonstration of N antibody in elk (Cervus elaphus) and mule deer (Odocoileus hemionus) prior to the epizootic in cattle and horses suggests that an enzootic cycle may exist in Colorado. PMID- 3035242 TI - Parasites, diseases and health status of sympatric populations of sambar deer and white-tailed deer in Florida. AB - From December 1983 to December 1984 a study on parasites, diseases and health status was conducted on sympatric populations of sambar deer (Cervus unicolor) and white-tailed deer (Odocoileus virginianus) from St. Vincent Island, Franklin County, Florida. Ten sambar and six white-tailed deer were examined. White-tailed deer had antibodies to epizootic hemorrhagic disease virus and bluetongue virus. Serologic tests for antibodies to the etiologic agents of bovine virus diarrhea, infectious bovine rhinotracheitis, vesicular stomatitis, parainfluenza 3, brucellosis, and leptospirosis were negative in both species of deer. White tailed deer harbored 19 species of parasites; all were typical of the parasite fauna of this species in coastal regions of the southeastern United States. Sambar deer harbored 13 species of parasites, which apparently were derived largely from white-tailed deer. The only exception was Dermacentor variabilis which occurs frequently on wild swine on the island. The general health status of sambar deer appeared to be better than that of white-tailed deer. This was hypothesized to result from the sambar deer's utilization of food resources unavailable or unacceptable to white-tailed deer and to the absence and/or lower frequency of certain pathogens in sambar deer. PMID- 3035243 TI - Prevalence of poxvirus in a population of Merriam's wild turkeys in Oregon. AB - An introduced population of Merriam's wild turkeys (Meleagris gallopavo) was examined for poxvirus when birds were trapped from January through April in 1981 and 1982. Poxvirus lesions were found in three of 113 (2.6%) turkeys. All infected birds were immature males. PMID- 3035244 TI - An unusual pleomorphic sarcoma in a hybrid mallard. AB - An unusual pleomorphic sarcoma from a hybrid mallard (Anas platyrhynchos) is described. Rhabdomyosarcoma was considered in the original differential diagnoses but rejected due to lack of specific characteristics generally seen in these tumors. The histologic characteristics described are consistent with mammalian sarcomas recorded in the literature as malignant fibrous histiocytoma. PMID- 3035245 TI - Intravitreous ganciclovir for patients receiving zidovudine. PMID- 3035246 TI - Ganciclovir treatment of cytomegalovirus disease in transplant recipients and other immunocompromised hosts. AB - Thirty-one immunocompromised patients with severe cytomegalovirus (CMV) disease were treated with intravenous ganciclovir. Twenty-one patients had received transplants--15 bone marrow recipients, five renal allograft recipients, and one liver transplant recipient--while the other ten were immunocompromised due to acquired immunodeficiency syndrome (six), hematologic malignancies (three), and systemic lupus erythematosus (one). They presented with one or more of the following syndromes: CMV pneumonitis (19), CMV of the gastrointestinal tract (six), CMV retinitis (seven), and CMV hepatitis (three). Seventeen (55%) of 31 patients demonstrated clinical improvement during ganciclovir therapy, with the best response seen in the transplant recipients. Viremia ceased in 14 (93.3%) of 15 patients after a mean of 4.7 days of therapy; viruria ceased in eight (53.3%) of 15 patients after a mean of 11 days of therapy. Ganciclovir plasma concentrations at a dosage of 2.5 mg/kg/three times a day were as follows: mean peak, 16.04 mumol/L; mean trough, 2.38 mumol/L. Neutropenia occurred in 11 (35%) of 31 patients and in nine (60%) of 15 bone marrow transplant recipients. We conclude that ganciclovir exerted an antiviral effect against CMV and may play a role in the treatment of CMV disease in patients with depressed immunity, especially bone marrow and organ transplant recipients. PMID- 3035247 TI - Leads from the MMWR. B-virus infection in humans--Pensacola, Florida. PMID- 3035248 TI - Experimental rotavirus vaccine passes first test; eventual goal: immunize newborns against most prevalent cause of life-threatening diarrhea. PMID- 3035249 TI - Presynaptic alpha 2-adrenoceptor mediated regulation of norepinephrine release in perfused mesenteric vasculatures in young and adult spontaneously hypertensive rats. AB - The present study was designed to evaluate the role of the presynaptic alpha 2 adrenoceptor in the pathogenesis of hypertension. Norepinephrine overflow during sympathetic nerve stimulation and its changes by presynaptic alpha 2-adrenoceptor inhibition were examined in the perfused mesenteric vasculatures of young and adult spontaneously hypertensive rats (SHR) compared with age-matched Wistar Kyoto rats (WKY). Electrical sympathetic nerve stimulation caused significantly greater overflow of endogenous norepinephrine from the adrenergic nerve terminals in young SHR than in age-matched WKY. Yohimbine, an alpha 2-adrenoceptor blocking agent, facilitated norepinephrine overflow from the adrenergic nerve terminals. The effects of yohimbine on norepinephrine overflow and pressor responses to electrical nerve stimulation were less in young SHR than in age-matched WKY. Norepinephrine overflow in adult SHR was similar to that in adult WKY, and differences in the effect of yohimbine on norepinephrine overflow between SHR and WKY were not marked at this chronic stage. These results suggest that enhanced norepinephrine overflow in the mesenteric vasculatures can be observed only in young SHR; this may be due in part to an impaired negative feed-back mechanism on the nerve terminals by presynaptic alpha 2-adrenoceptors. PMID- 3035250 TI - [Endocrine function following low flow anesthesia and surgery]. PMID- 3035251 TI - [Effects of dopamine, dobutamine, and dibutyryl cyclic AMP on coronary circulation in dogs]. PMID- 3035253 TI - [Studies of superoxide dismutase in rat gastric mucosal damage (the 1st report)- influence by inhibitor of Cu, Zn-superoxide dismutase, diethyldithiocarbamate]. PMID- 3035252 TI - [Prognostic significance of pretreatment CT scans in supratentorial astrocytomas receiving radiotherapy]. AB - Pretreatment brain CT scans of 61 patients receiving radiation and chemotherapy for supratentorial gliomas (23 low-grade astrocytomas, 21 grade III astrocytomas and 17 glioblastomas) were reviewed, and their prognoses were analyzed. Almost all gliomas with no CEA (contrast enhanced area) were low-grade, and their prognosis was promising. Benign and malignant gliomas with regular CEA were found in equal proportions, and their prognosis was relatively good. A majority of cases with irregular or ring-like CEA were malignant, and their prognosis was extremely poor. PMID- 3035254 TI - [Growth rate of human colorectal cancer xenografted in nude mice]. PMID- 3035256 TI - Use of microplate cell culture and enzyme immunoassay in titration of serum neutralizing antibody against Hochi strain of serotype 4 human rotavirus. AB - The tube neutralization test read by enzyme immunoassay developed by Wyatt et al. (1983) for serotype determination of human rotavirus was modified so as to use stationary cultures of MA104 cells in a microtiter plate instead of roller tube cultures. Sera obtained from different age groups were titrated for neutralizing antibody against serotype 4 human rotavirus Hochi strain by this test and the results were compared with those obtained by the plaque neutralization test. There was a good correlation between the titers obtained by the two tests and the age distribution pattern of serotype 4 neutralizing antibody was similar to those of serotype 1 and 3 antibodies previously reported. PMID- 3035258 TI - Immunological studies of uveitis. 1. Immune complex containing herpes virus antigens in four patients with acute retinal necrosis syndrome. AB - Four patients with acute retinal necrosis syndrome were examined. Three of them showed elevated antibody titers to varicella-zoster virus during the convalescent stage, while one of them showed an elevated titer to herpes simplex virus type 1, especially in the intraocular fluid. Also, these patients had an elevated circulating immune complex in the acute phase. Western blotting demonstrated that the immune complexes contained varicella-zoster or herpes simplex type 1 antigen, corresponding to the elevated antibody titer to either virus. PMID- 3035255 TI - [Role of free radicals in the formation of stress ulcer in rats]. PMID- 3035257 TI - [Plasma cyclic AMP and GMP levels in essential hypertension: interrelationship among plasma cyclic nucleotides sympathetic activity and renin-angiotensin system]. PMID- 3035259 TI - Analysis of three salmonid herpesvirus DNAs by restriction endonuclease cleavage patterns. PMID- 3035260 TI - Glioblastoma with intraocular metastasis in an old dog. PMID- 3035261 TI - Restriction endonuclease analysis of Aujeszky's disease viruses isolated in Japan. PMID- 3035262 TI - Existence of antigens cross-linking to tumor associated antigens of enzootic bovine leukosis in normal tissues of the bovine fetus. PMID- 3035263 TI - Translation of equine arteritis virus RNA in rabbit reticulocyte lysates. PMID- 3035265 TI - Plasmacytoma of a long clinical course with peripheral neuropathy and osteoblastic as well as osteolytic changes. PMID- 3035264 TI - Inhibition of human mammary carcinoma cell proliferation by retinoids and intracellular cAMP-elevating compounds. AB - Retinoids and cAMP-elevating agents markedly inhibited the proliferation of human mammary tumor cells. Their response has been previously correlated with the presence of estrogen receptor (ER) positivity. MDA-MB-231 cells were ER negative and insensitive to the antiproliferative effects of retinoids. However, their growth was markedly inhibited by agents that elevated intracellular cAMP levels, i.e., 8-bromo-cAMP, cholera toxin (CT), forskolin, and the phosphodiesterase inhibitor papaverine. The CT and forskolin inhibition of the ER-positive cells (MCF-7) was associated with an elevation of adenylate cyclase activity and intracellular cAMP levels; however, similar elevations in intracellular cAMP levels were not observed following CT or forskolin inhibition of MDA-MB-231 cells but only following the addition of the phosphodiesterase inhibitor 3-isobutyl-1 methylxanthine. PMID- 3035266 TI - [Optimal intensity and duration of training and the duration of walks during the after-care of myocardial infarct patients at a sanatorium]. AB - Daily exercise drills producing a heart rate increment of about 31.5% of the one recorded at first stress tolerance test and lasting for 105 min, in combination with daily 75 minutes' walks were not sufficient to increase stress tolerance. More intensive drills, producing a 50.6% heart rate increment, while lasting less than 1 hour per day, in combination with 2.5 hours' daily walks, resulted in a significant stress tolerance increase. In neither of the two groups was it associated with any significant rise of the double product, implicating extracardiac factors as the principal contributors. PMID- 3035268 TI - Effects of hydrochlorothiazide on Na-K-ATPase activity along the rat nephron. AB - Na-K-ATPase activity was determined in seven nephron segments of five-week-old, spontaneously hypertensive rats (SHR) with or without continuous hydrochlorothiazide (HCTZ) treatment for seven days. For comparison, the effects of HCTZ treatment on Na-K-ATPase activity in the nephron segments of age-matched normotensive Wistar-Kyoto rats (WKY) were also determined. Na-K-ATPase activity in proximal convoluted tubule (PCT), medullary thick ascending limb (MTAL), cortical thick ascending limb (CTAL), distal convoluted tubule (DCT) and cortical collecting duct (CCD) was significantly lower in HCTZ-treated SHR compared to control (untreated) SHR. However, there was no significant difference in Na-K ATPase activity in proximal straight tubule (PST) and medullary collecting duct (MCD) between HCTZ-treated and control SHR. HCTZ treatment also produced a significant decrease in blood pressure (BP) and creatinine clearance (CCr) in SHR. On the other hand, HCTZ treatment did not produce a significant change in Na K-ATPase activity in PCT, PST, MTAL, CTAL and MCD, in BP or in CCr in WKY. However, HCTZ treatment produced a decrease in the enzyme activity in the DCT and an increase in the enzyme activity in the CCD in WKY. The decrease in Na-K-ATPase activity in almost all nephron segments from SHR may be due to a significant decrease in CCr produced by HCTZ. On the other hand, a decrease in Na-K-ATPase activity in the DCT with an increase in the enzyme activity in the CCD from WKY suggest that renal compensation to the natriuretic effect of HCTZ occurs by an increase in Na+ reabsorption in the CCD. PMID- 3035267 TI - Mechanism of decreased vascular response to angiotensin II in renal vascular hypertension. AB - Compared to many forms of hypertension, vascular reactivity to angiotensin II (AII) is decreased in the early phase of two-kidney, one clip (2-K, 1C) renovascular hypertension (RVH). To determine the role of the AII receptor, we examined vascular responsiveness and 125I-AII binding to mesenteric artery membrane fractions after three weeks of 2-K, 1C RVH. Systolic blood pressure was 165 +/- 8 in RVH and 105 +/- 4 mm Hg in controls, P less than 0.001. Plasma renin activity was 4.2 +/- 0.6 in RVH and 1.4 +/- 0.5 ng AI/ml/hr in controls, P less than .001. The pressor response to exogenous AII was reduced by 40% in RVH. Since administration of a single dose of converting enzyme inhibitor (CEI) did not normalize the response to exogenous AII, the decreased reactivity was not caused by receptor occupancy. 125I-AII binding to mesenteric arteries was equal to or greater in RVH than controls at all concentrations of AII. Scatchard analysis revealed an increase in the total number of binding sites (BMAX): 140.8 +/- 6.3 in RVH versus 91.7 +/- 6.5 fmol/mg in controls, P less than 0.01, while the apparent dissociation constant was unchanged. To determine if the increase in circulating AII caused these binding alterations, rats were either treated with CEI for three days; or unilaterally nephrectomized. Both of these manipulations reversed the decrease in vascular responsiveness as well as the increase in receptor number (BMAX = 136 +/- 13.4 in RVH vs. 94 +/- 9.4 fmol/mg in RVH + nephrectomy, P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3035269 TI - [Nocturnal changes in cortisol, catecholamines, cAMP and histamine in children with nocturnal bronchial asthma and in healthy children]. AB - We analyzed the concentration of plasma epinephrine, norepinephrine, dopamine and C-AMP and urinary cortisol, histamine and creatinine in 28 children over a period from 4 p.m. to 8 a.m. 15 children were suffering from nocturnal asthma, 13 had no asthma problems. The cortisol-levels of both, the asthmatic and the healthy children were almost identical, the minimum cortisol-concentrations were found at 8 p.m. The concentration of plasma epinephrine in the asthmatic children was significantly higher than in the control group. Both groups showed minimum levels at 12 p.m. Plasma norepinephrine showed a continuous course with a raise in the early morning hours in both groups. Children without asthma had a continuous decrease of norepinephrine during the sleeping period and an increase in the early morning. Dopamine was significantly lower in the asthmatics. C-AMP was also on a lower level, but the difference between both groups was not significant. Histamine showed an extreme variability in the asthmatic children and was on higher levels during the whole period compared to the control group. The raised epinephrine levels in children with nocturnal asthma could be understood as compensation to a dysfunction of the receptor-transducer-adenylcyclase-complex of beta 2-receptors. A reduction of beta 2-receptors by anti-asthmatic therapy could also be responsible for these epinephrine levels. The significant difference in dopamine levels between children with and without nocturnal asthma is poorly understood. The increase of epinephrine and the decrease of dopamine could be caused by a modulated synthesis of catecholamines in children suffering from nocturnal asthma. PMID- 3035270 TI - Evidence for different immune responses to HIV in CSF and serum? AB - A case of acquired immune deficiency syndrome (AIDS) and a case of AIDS-related complex (ARC) are described. In both instances comparative Western blot analysis of cerebrospinal fluid (CSF) and serum samples show evidence of qualitative differences in antibody-binding patterns to viral polypeptides. PMID- 3035271 TI - Transmembrane signaling mechanisms of growth factors in Swiss 3T3 fibroblasts. PMID- 3035273 TI - Mousepox in inbred mice innately resistant or susceptible to lethal infection with ectromelia virus. I. Clinical responses. AB - Clinical responses to infection with ectromelia virus strain NIH-79 were determined in several strains of inbred mice. All mice were equally susceptible to infection, but mortality was strain dependent. BALB/c AnNCr, A/JNCr, DBA/2NCr and C3H/He/NCr MTV- mice were highly susceptible to lethal infection whereas AKR/NCr and SJL/NCr mice were moderately susceptible and C57BL/6NCr mice were highly resistant. Death rates were influenced strongly by virus dose and by route of inoculation. High doses were associated with early and high mortality. For a given dose, intraperitoneal inoculation resulted in the highest mortality and death rates were progressively reduced in mice inoculated by the footpad, subcutaneous and intranasal routes. Footpad swelling was prominent in resistant mice and in survivors among susceptible strains. Deaths among AKR and SJL mice were sporadic and often occurred late irrespective of virus dose. It is suggested that this pattern could be influenced by secondary contact infections or by immunologic injury associated with host responses to ectromelia virus. PMID- 3035272 TI - [Status of alpha 1-adrenergic regulation of stroke volume during hypokinesia in the rat]. AB - The positive effect of phenylephrine (PE) on stroke volume (SV) was 3 to 5 times weaker in the rats exposed to hypokinesia for 30 days as compared to the controls. After obsidan blockade of beta-adrenoreceptors SV increased in both groups and the intergroup differences in the PE effect remained significant but less pronounced. This can be attributed to a greater effectiveness of PE after obsidan administration during hypokinesia. Correlation analysis showed that the weak effects of PE on SV were potentiated by obsidan (potentiation was the stronger the weaker the effects) while the distinct effects were on the contrary inhibited. This demonstrates the synergy--antagonism relationships in the PE and obsidan interaction and seems to indicate that the common site of their action is alpha 1-adrenoreceptors. After phentolamine injections the PE effect on SV was not found. In this situation SV decreased; the SV decrease as well as its increase under the action of obsidan was less significant in the hypokinetic than in the control rats. The above investigation suggests that the activity of alpha 1-adrenoreceptors involved in the actualization of positive effects of agonists on SV is considerably lower during hypokinesia. PMID- 3035274 TI - Mousepox in inbred mice innately resistant or susceptible to lethal infection with ectromelia virus. II. Pathogenesis. AB - The pathogenesis of mousepox due to infection with ectromelia virus strain NIH-79 was characterized in genetically susceptible (BALB/cAnNCr) and genetically resistant (C57BL/6NCr) mice. BALB/c mice inoculated subcutaneous (s.c.) or intranasally (i.n.) had high mortality. Most mice died within 7 days from severe necrosis of the spleen and liver. Necrotic foci in livers of BALB/c mice that survived beyond 7 days often were accompanied by mononuclear cell infiltrates and by hyperplasia of lymphoid tissues. C57BL/6 mice inoculated by either route remained asymptomatic and necrotic lesions were mild or absent, whereas focal non suppurative hepatitis and lymphoid hyperplasia were prominent. Infectious virus and viral antigen were distributed widely in tissues of BALB/c mice, but had limited distribution in C57BL/6 mice. Both mouse strains had infection of the respiratory tract, genital tract, oral tissues and bone marrow, and BALB/c mice also had infection of the intestines. Both strains also developed serum antibody to vaccinia virus antigen after infection. The results show that ectromelia virus occurs in tissues conducive to mouse to mouse transmission and that the severity and character of mousepox lesions correlate directly with resistance and susceptibility to infection. They also support the concept that cellular immunity contributes to survival from infection. PMID- 3035275 TI - Mousepox in inbred mice innately resistant or susceptible to lethal infection with ectromelia virus. III. Experimental transmission of infection and derivation of virus-free progeny from previously infected dams. AB - The incidence and duration of transmission of infection with ectromelia virus strain NIH-79 was tested in innately resistant (C57BL/6) and innately susceptible (BALB/c) inbred mice. Transmission by C57BL/6 index mice occurred through 3 weeks and by BALB/c index mice through 4 weeks, although the duration of infection in individual index mice was often shorter. Soiled caging that previously housed infected mice was inconsistently infectious. Transmission was high in cages where infected mice died and were cannibalized by cagemates, but was low to moderate in cages where there was no cannibalism. Infected mice that were bred 6 weeks after they were infected, delivered virus-free progeny and did not transmit infection to their non-immune breeding partners. Sentinel mice housed in the room with experimentally infected mice were seronegative for antibody to ectromelia virus and to other murine viruses. These results support the view that infection with NIH-79 virus is typically short-lived. They also indicate that breeding of recovered mice can save valuable colonies that have been exposed to ectromelia virus. PMID- 3035276 TI - Observations on the replication of ectromelia virus in mouse-derived cell lines: implications for epidemiology of mousepox. AB - Ectromelia virus was shown to replicate in vitro in all lymphoma cell lines and in a small proportion of hybridoma lines tested. It was demonstrated that certain hybridoma cell lines, which were passed in ectromelia virus-infected mice, yielded ectromelia virus infectivity on explantation into tissue culture. This finding further substantiated the belief that ascitic fluid and hybridoma cell lines exposed to virus during mouse-passage could be important in the epidemiology of mousepox. PMID- 3035277 TI - Host age and genotypic effects on enterotropic mouse hepatitis virus infection. AB - Intestinal lesions due to infection with an enterotropic strain of mouse hepatitis virus (MHV-Y) were found to be more severe and wide-spread in BALB/cByJ and Cr1:CD-1(ICR) mice than in SJL mice inoculated at 1 week of age, using nonparametric ranking analysis. Lesions and viral antigen were limited largely to the bowel, but also occurred in the liver and brain of some mice. BALB/cByJ mice developed a particularly high prevalence of brain infection, resulting in mortality after the enteric phase of infection had ceased. MHV-Y antigen was present in neurons, glia and vascular endothelium in a vascular distribution. Cr1:CD-1(ICR) pups inoculated with MHV-Y at 4 or 7 days of age developed severe typhlocolitis, enteritis and encephalitis with moderate mortality. Pups infected at 2 or 3 weeks of age had mild intestinal lesions with minimal alteration of mucosal architecture, no encephalitis and no mortality. These results demonstrate that host age and genotype influence the course of enterotropic mouse hepatitis virus, as has been shown previously with non-enterotropic, respiratory-type strains of mouse hepatitis virus. PMID- 3035278 TI - Effect of mouse hepatitis virus infection on combination therapy of P388 leukemia with cyclophosphamide and pyrimidinones. AB - At least three marked differences were noted in the results compared from two parallel experiments using identical protocols with virus-free mice and mouse hepatitis virus (MHV) infected mice inoculated with P388 leukemia. First, the therapeutic effect of cyclophosphamide (CY), a cytotoxic antitumor drug, was apparently augmented in MHV-infected mice. A 162% increase of life span (ILS) was obtained in MHV-infected mice compared to a 100% ILS in uninfected mice. Second, the experimental error in terms of the range of animal survival time was much larger with MHV-infected mice than with uninfected mice. In MHV-infected mice, the therapeutic effect of the combination treatment with CY and pyrimidinone was not statistically different from that of the treatment with CY alone. In uninfected mice, the effects of the combination therapy at all doses of pyrimidinone were statistically more effective than that of CY treatment alone. PMID- 3035279 TI - A quantitative immunofluorescence test for detection of serum antibody to Sendai virus in mice. AB - A simple, semi-automated immunofluorescence assay, (TRACK XI), was developed for the detection and quantitation of circulating antibodies to Sendai virus in mice. The assay was validated by selecting Sendai virus-free and naturally infected mice from six different colonies and testing each serum in both ELISA and quantitative immunofluorescence (QIF) assays. The QIF test utilizes Sendai viral proteins immobilized within a dried colloid gel and permits serum antibody quantitation in 30 minutes. Using a dedicated fluorometer, antibody titers in test sera are calculated automatically from a three-point best fit calibration line. The QIF test gave 92.9% agreement with the ELISA and proved to be a reproducible, accurate and convenient assay for the quantitative measurement of serum antibody to Sendai virus in mice. PMID- 3035280 TI - Effects of leukotrienes B4, C4, and D4 on rat mesenteric microcirculation. AB - Arachidonic acid is metabolized to leukotriene (LT) B4, C4, D4, and E4 by lipoxygenase. LTB4 is a chemotactic agent while LTC4 and LTD4 stimulate smooth muscle fibers to contract. Mesenteric vessels have the capacity to release leukotrienes. The possibility that leukotrienes might be responsible for or contribute to mesenteric ischemia during mesenteric low flow, embolism, and thrombosis prompted us to investigate their action on mesenteric vessels. LTB4, C4, and D4 were applied topically on small bowel mesentery of 22 Sprague-Dawley rats in sequentially increasing concentrations. Mesenteric arterioles with diameters of 8-20 microns were observed through a microscope and vessel diameters were measured using a video shear monitor. LTB4 had no effect on diameter, but doses as low as 3 X 10(-8) M induced white blood cell adherence to venular endothelium, reflecting the potent chemotactic properties of this compound. LTC4 and D4 had no effect on systemic blood pressure or white blood cell adherence. Applications of 6.4 X 10(-9), 3.2 X 10(-8), and 6.4 X 10(-8) M LTC4 decreased mesenteric arteriolar diameter to 85.3* +/- 4.7% (mean +/- SD), 75.7* +/- 7.5%, and 66.8* +/- 6.1% of baseline, and 4 X 10(-9), 2 X 10(-8), and 4 X 10(-8) M LTD4 decreased diameter to 84.9* +/- 6.1%, 75.1* +/- 4.2%, and 64.1* +/- 5% of baseline, respectively (P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3035282 TI - Aldosterone production after short-term culture of rat adrenal capsule: responsiveness to angiotensin II, potassium and ACTH. AB - In vitro studies of aldosterone production have traditionally used freshly isolated adrenal glomerulosa tissue. In the present study we examined the effects of short-term culture of rat adrenal capsule on its secretory capacity by measuring both basal and stimulated aldosterone production. Capsules were maintained in culture for 24 h, and then responses to administered angiotensin II (1 X 10(-7) M), potassium (an increase of 2mM) and ACTH (1 X 10(-8) M) were determined during perifusion. Results were compared with responses by freshly isolated adrenal capsule. Although short-term culture reduced basal aldosterone production, responsiveness to administered stimuli was intact and often was greater than that observed with fresh capsular tissue. The results indicate that short-term culture of zona glomerulosa provides a suitable in vitro preparation for examining aldosterone secretory responsiveness to stimuli. PMID- 3035281 TI - Prostaglandin inhibition of acid is cAMP dependent. AB - Prostaglandin E (PGE) potently inhibits acid secretion stimulated by histamine, but not by acetylcholine or gastrin, and is accompanied by decreased intracellular cAMP. Adenylate cyclase receptor systems are composed of three complex proteins: cell receptor, nucleotide binding protein, and the catalytic subunit. The exact mechanism of PGE interaction with this complex remains unclear and elucidation of this site of action is the purpose of this study. We utilized molecular probes directed at the various components of adenylate cyclase. Cholera toxin alters the stimulatory subunit of the nucleotide binding proteins (Ns), rendering it resistant to normal deactivation, whereas N-ethylmaleimide (NEM) blocks the inhibitory subunit (Ni). Forskolin acts as a direct activator of the catalytic subunit of adenylate cyclase and 8-bromo-cAMP acts as a cyclic AMP mimetic. We measured in vitro acid secretion in isolated parietal cells by the assessment of [14C]aminopyrine (AP) accumulation. The PGE1 analog (miso) and the PGE2 analog (DMPG) were incubated in graded doses (10(-11) to 10(-6) M) with histamine (10(-6) M). Miso (10(-7) M) reduced AP accumulation to 21 +/- 8% of histamine alone (100%) and DMPG (10(-6) M) reduced AP to 61 +/- 9% (P less than 0.005 for both). AP accumulation stimulated by 8-Br-cAMP (10(-6) M) and forskolin (10(-6) M) was not significantly affected by either PGE analog (P greater than 0.05) suggesting that the site of PGE interaction is proximal to the activation of the catalytic subunit.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3035283 TI - The cholecalciferol sulphate system in mammals. AB - 7-Dehydrocholesterol sulphate has been identified in human and rat skin. The compound was isolated by anion exchange chromatography and following hydrolysis it was characterized by high-performance liquid chromatography and gas chromatography-mass spectrometry. Experiments with rats showed that 7 dehydrocholesterol sulphate can serve as a precursor of cholecalciferol sulphate and 25-hydroxy-cholecalciferol 3-sulphate, the latter compound being present in significant amounts in human blood. The sulphated sterols identified represent a previously unknown secosteroid system in mammals. PMID- 3035284 TI - The influence of plasma lipoproteins on steroidogenesis of cultured ovine fetal and neonatal adrenal cells. AB - The present study examined the effects of serum and lipoproteins on the function of cultured adrenal cells from 115-127-day-old ovine fetuses and from newborn lambs. On day 1 of culture, corticosteroid output was similar in medium containing 2% horse serum or in serum-free medium, both for fetal and neonatal cells. However, on day 5, cells cultured in the absence of serum produced smaller amounts of these steroids than cells maintained in medium containing serum; the difference was more marked under ACTH1-24 stimulation. Conversely, cAMP production was never lower in the absence than in the presence of serum. When stimulated by ACTH1-24 on day 2 of culture, fetal or neonatal adrenal cells incubated in the presence of a saturating concentration of ovine LDL produced more corticosteroids than cells incubated in serum-free medium; HDL also enhanced ACTH1-24-induced steroidogenesis, but to a lesser extent. VLDL was effective only with neonatal cells. In fetal and neonatal cells cultured for 6 days in ACTH-free medium, VLDL and LDL increased ACTH-induced steroidogenesis, but HDL did not. On the other hand, when cells were cultured in the presence of ACTH1-24, LDL and HDL were equipotent in supporting ACTH1-24-induced steroid output. Three major lipoprotein fractions were observed in serum of fetal and newborn lambs. The concentration of cholesterol was very low in the VLDL fraction of fetuses, but it was similar to that of newborns in LDL. Conversely, 4 times more cholesterol was present in HDL of newborns than in HDL of fetuses. These results suggest that: (i) after several days of cell culture, cholesterol availability is an important limiting factor for the steroidogenesis of cells maintained under serum-free conditions; (ii) both an "LDL pathway" and an "HDL pathway" are operating in adrenal cells from fetal as well as newborn sheep; (iii) LDL and HDL are important physiological sources of cholesterol to support steroidogenesis by fetal and neonatal adrenal cells. PMID- 3035285 TI - Adrenocortical function in 4-APP treated rats: a coupled stereological and biochemical study. AB - Adrenocortical function in 4-APP-induced (4-aminopyrazolo[3,4-d]pyrymidine) lipoprotein-deficient rats was studied in relation to quantitative morphologic changes in the gland. 4-APP treatment results in enlargement of the adrenal cortex and its zona fasciculata and reticularis cells. In enlarged livers, cholesterol and free fatty acid concentrations were similar to that of control rats, however a marked accumulation of triglycerides with a concomitant drop in hepatic delta 4-steroid hydrogenase activity was found. A profound drop in serum cholesterol in both, high and low density lipoproteins, as well as triglycerides and plasma corticosterone concentrations was accompanied by a marked lowering of cholesterol and corticosterone concentration in the adrenal gland. Corticosterone output by adrenal homogenates was higher in 4-APP treated rats than in control animals. Such a treatment did not change cholesterol side-chain cleavage, 11 beta hydroxylase, 3 beta-ol dehydrogenase-isomerase, steroid 5 alpha-reductase and neutral lipase activities when expressing results per unit weight of tissue or protein. However, when calculating per adrenocortical cell, adenine analogue applied increased 11 beta-hydroxylase, steroid 5 alpha-reductase and neutral lipase activities. Thus, coupled biochemical and stereologic studies revealed a complex and multidirectional effect of 4-APP on the rat adrenal cortex. This effect may be caused by serum lipoprotein deficiency and by toxic and stressful action of the adenine analogue on the rat. Also a direct effect of 4-APP on rat adrenal cortex may not be excluded. PMID- 3035288 TI - Characterization of steroid receptors in the gut and kidney of the frog (Rana catesbeiana) and in the gut of the turtle (Chrysemys picta). AB - Paraglucocorticoid- and paramineralocorticoid-binding cytosolic receptors (pGR, pMR) were demonstrated in the intestine and kidney of the frog, Rana catesbeiana and in the intestine of the turtle, Chrysemys picta, in the presence of sodium molybdate. These receptors were of high affinity and low capacity with the following binding parameters: pGR:Kd:frog intestine (FI), triamcinolone acetonide (TA): 3.3 nM, corticosterone (B): 3.4 nM; frog kidney (FK), TA:4.3 nM, B: 9.3 nM; turtle intestine (TI), TA: 4.8 nM; Nmax: FI, TA: 357, B: 371; FK, TA: 301, B: 157; TI, TA: 350 fmol/mg protein. pMR:Kd: FI, aldosterone: 0.9 and 90 nM (biphasic curves); FK, aldosterone: 0.6 and 36 nM (biphasic curves); Nmax: FI, 13 and 147 fmol/mg protein; FK, 78 and 109 fmol/mg protein. The receptor had the following ligand affinities: pGR: FI and FK: triamcinolone acetonide greater than DOC greater than 11 beta-hydroxyprogesterone greater than progesterone greater than corticosterone greater than cortisol greater than aldosterone greater than 11-dehydrocorticosterone greater than 17 alpha-hydroxyprogesterone greater than cortisone; TI: triamcinolone acetonide greater than corticosterone greater than progesterone greater than DOC greater than cortisol greater than aldosterone; pMR: FI and FK: corticosterone greater than 11 beta-hydroxyprogesterone greater than aldosterone greater than triamcinoline acetonide = cortisol greater than DOC greater than 11-dehydrocorticosterone greater than progesterone greater than 17 alpha-hydroxyprogesterone greater than cortisone. Androgens, estrogens or 18 hydroxycorticosterone did not compete for binding in either tissue. The heat activated frog receptors did not bind to naked DNA, though the turtle receptor did. It was possible to show that cytosol receptor-ligand complexes from all tissues were bound by nuclear acceptor sites. On linear sucrose gradients, the FI TA-receptor complex sediments with a single peak (7.5S), the FK TA-receptor complex gave two peaks (8.0 and 4.4S) and the TI TA-receptor complex showed a single peak (9.0S). The hydrodynamic parameters of the pGR's were determined by gel exclusion on Sephacel S-300. The following results were obtained: Mr: FI, 265, 80, 40 kDa (multiple proteins); FK, 280, 60, 20 kDa (multiple proteins); TI, 366 kDa; Rs: FI, 6.9, 3.9 nm; FK, 6.9, 2.9 nm; TI, 7.6 nm; f/f0: FI, 1.6; FK, 1.6; TI, 1.6. It is suggested on the basis of the binding and hydrodynamic parameters that non-mammalian epithelia corticosterone receptors have undergone biochemical evolution from one class of vertebrates to another. PMID- 3035287 TI - Biosynthesis of (20S)-20 alpha-reduced steroids simultaneously with corticosteroids in primary cultures of newborn rat adrenocortical cells stimulated by ACTH. AB - Newborn rat adrenal cells in primary culture produce corticosteroid hormones and (20S)-20 alpha-reduced progesterone metabolites in amounts which depend on ACTH concentrations and stimulation time. Eight (20S)-20 alpha-reduced progesterone metabolites, including 18-hydroxy-(20S)-20 alpha-dihydroprogesterone, were identified by comparison of their data in high performance liquid chromatography and in gas chromatography-mass spectrometry to those of existing or newly synthesized reference steroids. Quantitative studies of individual steroid biosynthesis were also performed using high performance liquid chromatography and gas chromatography. Several experiments were made without ACTH and with different concentrations of ACTH for periods of more than 3 weeks. The importance of the two main steroidogenic pathways, corticosteroid biosynthesis and progesterone reductive metabolism was modified by ACTH stimulation of the cultured cells. The progesterone reductive metabolism, important without ACTH and in the first days of ACTH stimulation, was decreased by 6.6 mU of ACTH/ml or higher concentrations but remained active throughout the life span of the stimulated cell cultures. PMID- 3035286 TI - Glucocorticoid receptors bound to the antagonist RU486 are not downregulated despite their capacity to interact in vitro with defined gene regions. AB - Modulation of gene expression by glucocorticoids involves interaction of these hormones with an intracellular receptor followed by 'transformation' of the hormone-receptor complex into a nuclear binding form. The molecular basis for the antiglucocorticoid action of high-affinity steroid analogues such as RU486 remains controversial. The effects of dexamethasone and RU486 on in vitro and in vivo properties of the receptor were compared using human lymphoblastoid IM-9 cells. In these cells, RU486 fully antagonized the glucocorticoid-specific induction of 5'-nucleotidase activity by dexamethasone. In vitro, however, RU486 bound receptor could be transformed and shown to interact specifically with cloned DNA fragments containing glucocorticoid response elements. These fragments included one from the mouse mammary tumour virus and two from the human growth hormone gene. In vivo, RU486-bound receptor did not behave like dexamethasone bound receptor. While receptor downregulation, a property of the transformed receptor, was achieved by dexamethasone, this did not occur with RU486. Likewise, RU486 did not affect receptor half-life under conditions when this was shortened by dexamethasone. These seemingly contradictory results can be reconciled by proposing that receptor transformation by agonists involves dissociation of the receptor oligomer to reveal a DNA-binding site that pre-exists on this protein. Although cell-free receptor dissociation and therefore DNA binding can occur even when the receptor is bound to RU486, this steroid maintains receptors in the untransformed state in the intact cell and therefore behaves a glucocorticoid antagonist in vivo. PMID- 3035289 TI - Torre's syndrome in association with premalignant and malignant tumors and suspected familial polyposis. PMID- 3035290 TI - Preservation of "peripheral" benzodiazepine receptors: differential effects of freezing on [3H]Ro 5-4864 and [3H]PK 11195 binding. AB - The equilibrium binding constants of [3H]Ro 5-4864 (a "peripheral" benzodiazepine receptor ligand) to renal membranes preserved by various freezing techniques were investigated. The Bmax for [3H]Ro 5-4864 binding to membranes from kidneys preserved as unwashed homogenates stored at -80, -20, or 5 degrees C, whole kidneys stores at -20 or 5 degrees C or as washed homogenate stored at -20 degrees C was significantly decreased (approximately 35%). Only when kidneys were frozen intact (using a dry-ice/acetone slurry) and stored at -80 degrees C was the density of [3H]Ro 5-4864 binding unchanged. However, the Bmax of [3H]PK 11195 (a putative "peripheral" benzodiazepine receptor antagonist) binding to renal membranes was unchanged following storage techniques that reduced the density of [3H]Ro 5-4864 binding 38%. No change was observed in the Kd values for [3H]Ro 5 4864 and [3H]PK 11195 binding to renal membranes preserved under any condition tested. These results demonstrate a method for the preservation of [3H]Ro 5-4864 binding to renal membranes, and suggests that [3H]Ro 5-4864 and [3H]PK 11195 bind to unique sites on or near the "peripheral" benzodiazepine receptor. [corrected] PMID- 3035291 TI - Regulation of proliferation of bone marrow-derived macrophages. AB - The rate of mobilisation of macrophages from marrow is an important determinant of the outcome of many pathological lesions. Studies of BMDM have provided insight into the complex feedback regulation that may control macrophage production. In addition BMDM possess several properties in common with transformed cells of other tissue origins and an understanding of their growth regulation may provide an insight into the mechanism of malignant transformation. PMID- 3035293 TI - Assessment of nerve damage in the feet of long-distance runners. AB - To answer the question, "Does long-distance running injure nerves of the feet and legs?" we invited the 25 members of the Rochester Track Club who had run the greatest number of miles in their lifetime to participate in a study that involved neurologic examination, determination of detection thresholds of touch pressure, vibratory, and cooling sensations of the foot, and evaluation of nerve conduction. None of the runners had clinical symptoms or signs of peripheral neuropathy. Most of them reported having had toe and foot injuries, sometimes associated with short-lived sensory symptoms. A computer-assisted sensory evaluation of the detection threshold for vibratory sensation of the toe revealed slightly, but significantly, higher values in the runners in comparison with age- and sex-matched control subjects. Similarly, small, but statistically significant, differences were also observed for some attributes of nerve conduction in the leg and foot nerves of the runners. These results suggest that long-distance running causes multiple small injuries to the toes and feet, which lead to measurable differences in the detection threshold for vibratory sensation and in nerve conduction but not to overt neuropathy. The trivial subclinical neuropathic deficits we noted are readily offset by the assumed health and recreational benefits of running. PMID- 3035292 TI - Sodium salicylate has no effect on cerebrospinal fluid [H+] in dogs with normal acid-base balance. AB - The experiments described here were designed to investigate the possibility that central stimulation of respiration by salicylates may be due to changes in [H+] of cerebral fluids. Two groups (n = 6 in each) of anesthetized, paralyzed, and mechanically ventilated dogs were studied for 6 hr. Renal pedicles were ligated to maintain blood salicylate level constant. Group II received 150 mg/kg Na salicylate intravenously at 0 hr after samples had been obtained. Group I (control) received equal volume of half-normal saline. Mean plasma salicylate levels were 18.9, 18.4, and 19.6 mg % at 0.5, 3, and 6 hr after administration of Na salicylate. Respective cisternal cerebrospinal fluid (CSF) levels were 3.2, 4.8, and 5.9 mg %. Salicylate-induced hyperthermia was prevented by peritoneal cold dialysis, and a rise in PaCO2 was prevented by increasing ventilation. During the 6 hr of relatively normal systemic acid-base balance, cisternal CSF mean PCO2 values were 45.3, 43.6, and 49.3 mm Hg at 0, 3, and 6 hr in the control group; in group II, respective values were 46.9, 45.7, and 47.7 mm Hg. Cisternal CSF [H+] were 44.4, 45.2, and 50.5 nEq/L in group I at 0, 3, and 6 hr. Respective values in group II were 45.0, 47.5, and 50.6 nEq/L. These values were similar and statistically insignificant from those in group I. In both groups cisternal CSF [HCO3-] fell about 2 and CSF lactate concentration rose about 1 mEq/L at 6 hr.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3035294 TI - Numerical grading of astrocytomas. AB - Ninety-three astrocytomas from biopsy material including glioblastomas ('astrocytomas grade 4') were graded according to Kernohan (1949) by light microscopy. Feulgen sections were subjected to an automated microscopic analysis to obtain morphometric-densitometric data of the tumour cell nuclei. These quantitative and reproducible data showed a significant correlation with malignancy expressed in terms of Kernohan and prove automated image analysis to be a valuable tool in the grading of gliomas. This is in particular true if the nuclear parameters determined by image analysis are completed by histologic features which were recorded semiquantitatively by subjective light microscopic evaluation as is usual in clinic pathologic diagnosis of brain tumours. A quadratic discriminant analysis of morphometric-densitometric data of tumour cell nuclei and semiquantitative microscopic data gave a 94% agreement with subjective grading. PMID- 3035295 TI - [The cotton rat (Sigmodon hispidus) as an experimental model for studying viruses in human respiratory tract infections. I. Para-influenza virus type 1, 2 and 3, adenovirus type 5 and RS virus]. PMID- 3035296 TI - Studies on the effects of oxamniquine on autonomic transmissions. AB - The effects of oxamniquine have been investigated on parasympathetic and sympathetic transmissions using in vitro and in vivo laboratory models. Oxamniquine (3.6 X 10(-5)-5.5 X 10(-4) M) depressed or abolished electrically evoked contractions of the isolated chick esophagus and isolated vas deferens muscle preparations. At the same concentration (3.6 X 10(-5)-5.5 X 10(-4) M) levels, the drug also reduced or abolished the indirect, electrically-provoked twitches of the rat isolated phrenic nerve-hemidiaphragm muscle preparations and contractions of the cat nictitating membrane induced by pre-ganglionic cervical sympathetic nerve trunk electrical stimulation in vivo. These results strongly suggest blockade of sympathetic and parasympathetic transmissions by oxamniquine. PMID- 3035297 TI - Naloxone-sensitive and GABAA receptor mediated analgesic response of benzodiazepines in mice. AB - Various benzodiazepines (BZs) were investigated for their analgesic effect by the tail-flick method in mice. Central type BZ-receptor agonists such as clonazepam, chlordiazepoxide and diazepam showed analgesia while the peripheral BZ-receptor binding agent, Ro 5-4864, and the BZ micromolar binding agent phenytoin, failed to show any effect. Other BZs, such as lorazepam and nitrazepam, were ineffective in producing analgesia. Pretreatment with naloxone antagonized the analgesic effect of central type BZ-receptor agonists. Similarly, pretreatment with Ro 15 1788, a central BZ-receptor antagonist, also blocked the analgesic effect. When the involvement of the GABAergic system in the analgesic response of BZ agonists was investigated, a potentiation of BZ action was seen as a combination of a subeffective dose of clonazepam with a subanalgesic dose of muscimol, a specific GABAA agonist, showed an enhanced effect. Moreover, pretreatment with GABAergic substances like pentobarbitone also showed a facilitatory effect on BZ-induced analgesia. On the other hand, a combination of clonazepam with baclofen, a specific GABAB agonist, failed to show any synergistic effect. The analgesic effect of central type BZ-receptor agonists was found to be bicuculline reversible. It is concluded that GABAA receptor activation has a modulatory role in the naloxone sensitive analgesic effect of central type BZ-receptor agonists. PMID- 3035298 TI - [Treatment with enalapril causing leukopenia with septicemia and affected liver]. PMID- 3035299 TI - Laryngeal papillomatosis: clinical, histopathologic and molecular studies. AB - The clinical course and pathology of 57 patients with laryngeal papillomatosis were reviewed. Tissues from 26 patients were analyzed for human papillomavirus (HPV) DNA by Southern blot hybridization. Histopathologic evaluation of the papillomas showed no correlation with age of onset or clinical pattern of remission and recurrence. The pathology was characterized by abnormal squamous maturation with parakeratosis, retardation of superficial cell maturation, papillomatosis, and basal hyperplasia. HPV DNA was present in all lesions, with 92% containing either HPV-6 or 11. Latent HPV DNA was detected in clinically uninvolved tissues of 11 of 14 (78.5%) patients studied. There was no correlation between HPV type, histopathology and/or clinical pattern. Despite homogeneity of pathology, the clinical expression of laryngeal HPV infection varied widely. A mechanism for the pathogenesis of laryngeal papillomatosis, based on the concept of maturational arrest, is proposed. PMID- 3035300 TI - [Cardiovascular function in liver cirrhosis]. AB - The hemodynamic pattern in patients with cirrhotic liver disease shows a hypercirculatory state, with elevated cardiac output and decreased systemic vascular resistance. Studies on myocardial function gave different results, whereby non hepatic factors as a cause of myocardial dysfunction are reasonable. Decreased vascular resistance is predominantly caused by an accumulation of vasodilating substances. A dysfunction of vasoconstricting systems could not be found. A previously discussed interference of the sympathetic nervous system could not be confirmed in further studies. This hypercirculatory state may be catastrophic in hypovolemic states, as in acute bleeding, or concomitant septic hyperdynamic states, because the initial compensatory mechanisms are not available any more. PMID- 3035301 TI - A second look at the second messenger hypothesis. AB - Several hundred hormones, neurotransmitters, growth factors and other "first messengers" bind to specific cell membrane receptors and induce a myriad of effects: short term, transport, metabolic, mitotic and regulation of thousands of specific genes. Yet, less than a dozen "second messengers" have been clearly established to date. Even allowing for the discovery of a large number of additional second messengers, there remains a paradox in terms of information transfer within the cell: how can so many specific signals produce so many effects through so few relatively nonspecific intermediates? We consider several possible solutions to this paradox, including the hypothesis that signal specificity is encoded in part in the primary structure of the receptor. PMID- 3035302 TI - Stimulation of insulin secretion by beta-endorphins (1-27 & 1-31). AB - Synthetic human beta-endorphin potentiates insulin secretion by the isolated perfused rat pancreas when glucose is present in the perfusate at concentrations of either 125 or 200 mg/dl, whereas it fails to exert any effect on insulin secretion in the presence of a substimulatory concentration of 100 mg/dl. Similar potentiation of insulin secretion occurred in response to the 1-27 fragment (beta endorphin1-27) of beta-endorphin. This transient potentiation lasts only 3 to 4 minutes, whereupon secretion returns toward control levels. Thus beta-endorphin produces only a transient spike-like secretory profile similar to the first phase of glucose-induced insulin secretion and it fails to produce any chronic insulin secretory response comparable to the second phase of insulin secretion. The insulinotropic effect of beta-endorphins occurred at concentrations varying from 0.1 to 5.0 ug/ml. These data suggest that beta-endorphin and beta-endorphin1-27 potentiate insulin secretion via a common beta cell opioid receptor, and that beta-endorphin may exert a paracrine control of insulin secretion. However, any such regulation appears to be via short-term alterations in the secretory process per se. PMID- 3035303 TI - Clinical and treatment effects on 3H-clonidine and 3H-imipramine binding in elderly depressed patients. AB - 3H-clonidine and 3H-imipramine binding were measured in depressed patients, 55 years and older. There was no significant difference in either 3H-clonidine or 3H imipramine binding between depressed patients and age- and sex-matched controls. There was no significant correlation between 3H-clonidine or 3H-imipramine binding and severity of depression before treatment. There was a significant negative correlation between the KD of 3H-imipramine binding sites and Hamilton score over seven weeks of antidepressant treatment. There was no significant difference between receptor data of responders and nonresponders to antidepressant treatment. PMID- 3035305 TI - Correlation between the enhancement of flunitrazepam binding by GABA and seizure susceptibility in mice. AB - Various populations of mice exhibit differential sensitivity to seizure-inducing agents. The relationship of seizure susceptibility to alterations in the GABA receptor complex was investigated in six different populations of mice consisting of four inbred strains (C57BL, DBA, C3H, and BALB) and two selected lines (long sleep and short sleep). Seizure activity was induced by intraperitoneal administration of the GAD inhibitor, 3-mercaptopropionic acid, and latencies to seizure onset and tonus were measured. In naive mice of the same populations, GABA enhancement of 3H-flunitrazepam binding was measured in extensively washed whole brain membranes at several GABA concentrations. Both differential seizure sensitivity to 3-mercaptopropionic acid and differential enhancement of 3H flunitrazepam binding by GABA were observed in these six populations of mice. Correlational analyses indicated a positive correlation between the degree of GABA enhancement of 3H-flunitrazepam binding and resistance to the seizure inducing properties of 3-mercaptopropionic acid. These data suggest that genetic differences in sensitivity to seizure-inducing agents that disrupt the GABAergic system may be related to differences in coupling between the various receptors associated with the GABA receptor complex. PMID- 3035304 TI - (+)- and (-)-N-allylnormetazocine binding sites in mouse brain: in vitro and in vivo characterization and regional distribution. AB - In vivo and in vitro binding studies, both in whole brain and in selected areas, indicate that non-identical (+)- and (-)-NANM sites exist in the mouse brain, and each exhibits a different regional distribution. The in vivo binding of (+)-3H NANM was found to be saturable at pharmacologically relevant doses, and represents a relatively small (10-22%) portion of total brain (+)-3H-NANM concentrations. The in vivo binding of (+)-3H-NANM was selectively displaced by (+)-NANM and PCP, and more sensitive to haloperidol and (+)-ketocyclazocine than the (-)-3H-NANM site. The in vivo binding of (-)-3H-NANM was selectively displaced by (-)-NANM, and more sensitive to naloxone and (-) ketocyclazocine than the (+)-3H-NANM site, and insensitive to PCP. This study indicates that the investigation of NANM binding sites is possible using in vivo binding techniques, and that each isomer apparently binds, in the mouse brain, to a single class of distinct sites. PMID- 3035306 TI - Effect of phencyclidine on post-tetanic twitch tension of the mouse diaphragm preparation. AB - The effects of phencyclidine(PCP) on the post-tetanic potentiation(PTP) of twitch tension were studied on the isolated mouse phrenic nerve diaphragm preparation. Phencyclidine increased directly elicited twitch tension while it decreased post tetanic potentiation of the indirectly elicited twitch tension. The maximal depression effect of the PTP was found after higher frequencies and longer durations of stimulation. After repetitive stimulation, the amplitude of endplate potential was potentiated. Phencyclidine decreased the post-tetanic potentiation of the amplitude of endplate potential while the quantal content of the endplate potential was not affected. 4-Aminopyridine increased both directly and indirectly elicited twitch tension while it did not inhibit the post-tetanic potentiation of the twitch tension. It is concluded that phencyclidine suppressed the post-tetanic potentiation of the indirectly elicited twitch tension. The depressant effect may be mainly due to its effect on the acetylcholine receptor ionic channel complex of the motor endplate. PMID- 3035307 TI - Effects of acute and chronic methylphenidate administration on beta-endorphin, growth hormone, prolactin and cortisol in children with attention deficit disorder and hyperactivity. AB - The effect of 5 mg/p.o. methylphenidate (MPH) challenge on beta-endorphin (beta EP), growth hormone (GH), prolactin (Prl) and cortisol was investigated in 16 children suffering from attention deficit disorder with hyperactivity (ADDH) before and after 4 weeks MPH treatment. The study population consisted of 13 males and 3 females aged 6-11 years. All patients were drug free for at least 3 months prior to investigation. The severity of ADDH symptomatology and response to MPH chronic treatment was assessed using parent/teacher abbreviated Conners rating scale. Blood samples for beta-EP, cortisol, Prl and GH were drawn before initiation of treatment (basal pre-treatment level), 2 hours after MPH challenge, 4 weeks after MPH treatment (basal post-treatment level) and 2 hours after re challenge with MPH. Chronic MPH treatment resulted in a decrease in basal Prl levels (5.5 +/- 2.8 vs 3.7 +/- 1.9 ng/ml; p less than 0.05). Pre-treatment challenge stimulates significantly both beta-EP (15.0 +/- 7.5 vs 12.5 +/- 5.3 pmol/l; p less than 0.05) and cortisol secretion (20.6 +/- 6.6 vs 12.6 +/- 5.8 micrograms/dl; p less than 0.05), and suppressed Prl secretion (4.0 +/- 1.5 vs 5.5 +/- 2.8 ng/ml; p less than 0.05). Re-challenge with MPH enhanced beta-EP levels (14.9 +/- 8.6 vs 10.6 +/- 5.0 pmol/l; p less than 0.05) but failed to affect cortisol, Prl and GH secretion. The acute and chronic neuroendocrine effects of MPH administration might be related to its dopaminergic and adrenergic agonistic activity. It might be that the stimulatory effect of single and repeated acute MPH administration on beta-EP release contributes to the beneficial effect of MPH treatment in ADDH children. PMID- 3035308 TI - Bivalent opioid peptide analogues with reduced distances between pharmacophores. AB - To investigate the role of distance between two opioid peptide pharmacophores on in vitro and in vivo activities, three new bivalent opioid analogues have been synthesized in which the dipeptide Tyr-D-Phe was connected with diamine moieties ("bridges"). The analogue with a hydrazine bridge has high receptor affinity to mu, kappa, and delta receptor types, as well as potent and long acting antinociceptive activity after intraperitoneal administration. PMID- 3035309 TI - Opioid control of oxytocin secretion: evidence of distinct regulatory actions of two opiate receptor types. AB - Stress induced oxytocin (OT) secretion was measured in female rats following treatment with various opiate antagonists selective for different types of opiate receptor. Naloxone (mu selective) and MR2266 BS (kappa selective) potentiated the OT response to an emotional stress (1 min. immobilization) whereas the delta selective antagonist ICI 154129 was without effect. Similarly, naloxone and MR2266 BS, but not ICI 154129, potentiated the response to a physical stress (i.p. hypertonic saline). A dose response comparison of the actions of naloxone and MR2266 BS revealed that naloxone was most effective in potentiating the immobilization response whereas MR2266 BS elicited greater responses than naloxone when administered prior to hypertonic saline. The results indicate that the opioid regulation of stress induced OT secretion is primarily mediated via mu and kappa opiate receptor types, the two types differentially regulating the OT response to two different stressors. PMID- 3035311 TI - Cyclic adenosine 3',5'-monophosphate response to dopamine in mouse lymphoid cells and cell lines. AB - Cyclic adenosine 3',5'-monophosphate responses to dopamine and isoproterenol were studied in mouse and rat spleen, thymus, lymph nodes and Peyer's patches lymphocytes and in 7 mouse cell lines of T- and B-lymphoid derivation. The responses of normal cells to dopamine were moderate, of the same extent, but selective to spleen and thymus in mouse, and to spleen and lymph nodes in rat. The YAC-1 T lymphoma cell line was sensitive to dopamine with a higher magnitude than normal lymphoid cells. Dopamine was less potent than isoproterenol in all cells, and whereas dopamine-sensitive and isoproterenol-sensitive cells, or dopamine-insensitive and isoproterenol-insensitive cells were found, no cell type was dopamine-sensitive and isoproterenol-insensitive. Altogether, these results suggest that only a small subset of lymphocytes is susceptible to the cAMP elevating action of dopamine. PMID- 3035312 TI - Presynaptic effects of glucocorticoids on dopaminergic and cholinergic synaptosomes. Implications for rapid endocrine-neural interactions in stress. AB - Synaptosomal preparations from rat hippocampus were incubated with methylprednisolone or adrenocorticotropin. High affinity choline uptake was not affected by either hormones. Methylprednisolone however enhanced newly synthesized acetylcholine release in the presence of high potassium or acetylcholine concentrations, while adrenocorticotropin had no effect. Dopamine uptake was inhibited when synaptosomes from septum or striatum were incubated with methylprednisolone. We conclude: a) high glucocorticoid concentrations and not adrenocorticotropin can directly enhance acetylcholine release but only from stimulated cholinergic synaptosomes, and b) high glucocorticoids can reduce dopamine uptake by dopaminergic synaptosomes. The results imply that increased glucocorticoid levels during stress or disease, can directly modulate the neuronal activity of specific cholinergic and dopaminergic systems in the brain. PMID- 3035310 TI - Dextrorphan: an antagonist for phencyclidine receptors. AB - Radio-binding assay, bioassay and HPLC detection were used to observe the antagonistic effects of dextrorphan on PCP's actions. Dextrorphan displayed high affinity to PCP receptor in the rabbit mesenteric blood vessels. It had weak PCP like bioactivity, but could antagonize PCP's action dose-dependently in vitro study with the rabbit ear artery preparation and shifted the dose-response curve of PCP to the right. After PCP administration, the content of norepinephrine in the vascular bath medium was increased, which was reversed by dextrorphan. Thus suggests that dextrorphan is an antagonist with very mild agonistic action for PCP receptors. PMID- 3035313 TI - Biphasic effects of dibutyryl cyclic adenosine 3',5'-monophosphate on synergistic stimulation of DNA synthesis by diacylglycerol, and the ionophore A23187 in guinea pig lymphocytes. AB - When guinea pig lymphocytes were cultured with 1-oleoyl-2-acetylglycerol (OAG) and the ionophore A23187 for 8 h, [3H]-thymidine incorporation into the acid insoluble fraction of the cells was stimulated synergistically. Further addition of dibutyryl cAMP caused a biphasic effect on the synergistic stimulation. Dibutyryl cAMP augmented the synergistic stimulation when A23187 was at the concentration of 0.075 micrograms/ml, but inhibited it when the ionophore was at 0.25 micrograms/ml. At the higher concentration of A23187, dibutyryl cAMP stimulated the [3H]thymidine incorporation when culture was for 4 h, but inhibited it when culture was for 8 h. The results were the same when 12-0 tetradecanoylphorbol-13-acetate (TPA) was used instead of OAG. Butyrate could replace dibutyryl cAMP for stimulation of [3H]thymidine incorporation in combination with TPA and A23187, but not with OAG and A23187 at the lower ionophore concentration. Dibutyryl cAMP but not butyrate stimulated ornithine decarboxylase induction caused by TPA and A23187. These results suggest that the effect of dibutyryl cAMP on DNA synthesis induced by OAG and A23187 was biphasic and depended on the concentration of A23187 and on the time of culture, and that the stimulation mechanism of butyrate is different from that of dibutyryl cAMP. PMID- 3035314 TI - [Radiography, ultrasonic diagnosis and radioimmunologic analysis in the differential diagnosis of diffuse goiter and thyroiditis]. AB - Radionuclide functional dynamic investigation of the thyroid with 99mTc pertechnetate was performed in 62 patients with diffuse euthyroid and toxic goiter, thyroiditides and in 17 controls. An analysis of the results of investigations and their comparison with clinical findings, an echo- and scintigraphic picture of the thyroid as well as with the levels of total thyroxine, triidothyronine and thyrotropin of the hypophysis showed that the results of a dynamic test of pertechnetate trapping corresponded, to a large extent, to the clinical status of patients with diffuse thyroid changes, correlated with an echographic picture and made it possible to define significant radiodiagnostic signs of thyroiditides. The proposed methods made it possible to increase the volume and improve the quality of diagnostic information in functional investigations of the thyroid status raising the efficacy of diagnosis of thyroid diseases and reducing radiation exposures and the time of investigations. PMID- 3035317 TI - [Scintigraphy of the sacroiliac joints in Bechterew's disease]. PMID- 3035315 TI - [Radionuclide visualization of non-skeletogenous tumors using 99mTc-phosphates]. AB - The paper is concerned with the results of radionuclide investigation of 51 patients with different tumors located beyond the osseous tissue. In spite of the fact that 99mTc-phosphates were well known as agents for radionuclide diagnosis of bone tumors, they can be employed for obtaining certain diagnostic information concerning topical diagnosis and the determination of a degree of the distribution of processes of nonskeletogenous nature. Of 51 patients with soft tissue pathology clear visualization of lesion foci was obtained in 34 patients (66.7%). Positive results of radionuclide diagnosis with 99mTc-pyrophosphate were noted in scintigraphy of patients with breast, ovarian, soft tissue and maxillofacial tumors. Besides, clear visualization of lesion foci was achieved in 6 of 11 patients with benign diseases: postoperative scars, inflammatory infiltrates, synovioma. The results obtained indicated the absence of necessary specificity of 99mTc-phosphates to osseous tissue. PMID- 3035318 TI - [Antibiotic resistance of Pseudomonas aeruginosa: a fact and a problem at the University Hospital Center of Cocody (Abidjan)]. AB - The in vitro activity of 4 beta lactam compounds (carbenicillin, ampicillin, cefotaxim, azlocillin), 4 aminoglycosides (gentamicin, dibekacin, netilmicin, amikacin), chloramphenicol and tetracyclin chlorhydrate was studied on 30 strains of Pseudomonas aeruginosa. Amikacin is the most efficient with 97% of sensitive strains, then dibekacin (80%), azlocillin (76%) and carbenicillin (73%). Gentamicin and netilmicin are the less efficient with respectively 43% and 63% resistant strains. Chloramphenicol, ampicillin and cefotaxim have not any activity on these strains. This study reveals that 3% of the strains are polyresistant. PMID- 3035316 TI - [Indirect radionuclide lymphography in the evaluation of the completeness of iliac lymphadenectomy and the diagnosis of postoperative lymphatic cysts in uterine cancer]. AB - Indirect radionuclide lymphography was performed in 152 women in the next 1-2 weeks after operation on the iliac lymph nodes. In 136 of them a radiopharmaceutical (198Au-colloid, 113mIn-coind, 113mIn-coinol) was accumulated in the inguinofemoral lymph nodes only, being in favor of radical iliac lymphadenectomy. X-ray lymphographic control confirmed in all cases that the iliac lymph nodes had been completely removed. Lymphatic cysts were detected in 16 out of 152 patients and verified in all the cases. Thus radiolymphography is indicated in early terms after lymphadenectomy for the assessment of its efficacy and diagnosis of lymphatic cysts. PMID- 3035319 TI - [Setting-up a system of health statistics collection in a rural environment in the Ivory Coast]. AB - In order to improve the sanitary data collection system from the rural Health Centers, Ivory-Coast has modified it in acting upon the three stages of collection. At the level of rural Health Centers, it has been created, from observations and data collected in a sample of 5 centers, a new classification of diseases including 128 items allowing all male and female nurses to classify 94% of the patients examined. At intermediary level, their monthly reports are collected and checked by the chief Medical Practitioner of the Rural Health Sector, who is responsible for all medicine not pertaining to hospital centers. At central level, computerised data treatment and exploitation make possible national data spreading and back-up during the third month after the writing of their report by the nurse. PMID- 3035320 TI - Feasibility study of imaging a living murine tumor by electron paramagnetic resonance. AB - An electron paramagnetic resonance image was measured for the first time from in vivo field gradient spectra of a living murine tumor (Cloudman S-91 melanoma in the tail of a DBA-2J mouse) using the paramagnetic nitroxide imaging agent 3 carboxamido-2,2,5,5-tetramethylpyrroline-1-oxyl injected into the tail vein. The experiments were accomplished at L-band frequency (1.55 GHz) with a single-turn flat-loop coil. A cross-sectional image was obtained perpendicular to the tail axis, which clearly distinguished features to the submillimeter resolution level. PMID- 3035321 TI - Effects of synacthen on lipid metabolism in the perfused epididymal fat pad of the rat. AB - Rats were treated with Synacthen, a synthetic corticotrophin analogue, to induce hypercorticism. The epididymal fat pad was selectively cannulated and perfused. In fasted rats acetone ether powder lipoprotein lipase (LPL) activity rose during treatment to levels found in fed controls. In fed animals no further rise in LPL activity was observed during Synacthen treatment. However, the heparin-elutable LPL activity did not change during this treatment in fasted nor fed animals. Pharmacologic levels of insulin in the perfusion medium caused an increase in heparin-releasable LPL activity as a percentage of total fat pad LPL activity (15% v 48%). Hydrolysis of chylomicrons was higher in fasted three days treated animals then in controls (10 +/- 4% v 2 +/- 2%). In this group a higher uptake of liberated free fatty acids was found (2.6 +/- 1.5% v 1.0 +/- 0.5% in controls). The increase in hydrolysis rate and uptake of fatty acids in the treated fasted animals could not be explained by an increase in releasable LPL activity. Fatty acid release from the fat pad was lower in treated animals than in controls (fasted and fed), basally as well as after adrenalin stimulation. The observation that the epididymal fat pad retains its weight during hypercorticism may therefore be ascribed to an increased influx of fatty acids from increased hydrolysis of TG-rich particles and to an inhibited efflux of fatty acids from the adipocyte. The discrepancy between the LPL activity extractable from an acetone ether powder and the heparin releasable LPL activity suggests impairment of the transport of LPL from the adipocyte to the heparin releasable pool at the endothelium. PMID- 3035323 TI - Purification and assay of calspermin: a heat-stable calmodulin-binding protein in male reproductive system and central nervous system. PMID- 3035322 TI - Isolation and characterization of calmodulin-dependent myosin heavy chain kinase from intestinal brush border. PMID- 3035324 TI - The calmodulin gene of Drosophila melanogaster. PMID- 3035325 TI - Calmodulin from Trypanosoma brucei: immunological analysis and genomic organization. PMID- 3035326 TI - The use of synthetic oligodeoxyribonucleotides in the examination of calmodulin gene and protein structure and function. PMID- 3035327 TI - Cloning and evolution of calcium-dependent protease, cDNA cloning of a novel family of calcium-binding proteins. PMID- 3035328 TI - Production and characterization of a monoclonal antibody cross-reactive with calmodulin, calmodulin-dependent phosphodiesterase, and protein phosphatase. PMID- 3035329 TI - Methods for analyzing bovine papilloma virus-based calmodulin expression vectors. PMID- 3035330 TI - Purification of novel calcium-binding proteins from bovine brain. PMID- 3035331 TI - Reconstitution of calmodulin-sensitive adenylate cyclase from bovine brain with phospholipids, calmodulin, and beta-adrenergic receptors. PMID- 3035332 TI - Calmodulin-dependent protein phosphatase: isolation of subunits and reconstitution to holoenzyme. PMID- 3035333 TI - Multivalent regulation of the nusA operon of Escherichia coli. AB - The rate of synthesis and intracellular content of the NusA protein, a transcription termination factor, were determined for wild-type and nusA and/or nusB mutants of Escherichia coli. Both the rate and content of NusA in wild-type strains were similar to that of the RNA polymerase sigma subunit, a transcription initiation factor, on a molar basis, and about 30%-40% the levels of RNA polymerase beta beta' subunits. At the stationary phase of cell growth, the values increased in parallel for both transcription factors up to approximately the level of the beta beta' subunits. In nus mutants, the rate of synthesis and the content of the sigma subunit were significantly increased. These observations together suggest that the two transcription factors are coordinately regulated. PMID- 3035335 TI - Tn1545: a conjugative shuttle transposon. AB - Tn1545, from Streptococcus pneumoniae BM4200, confers resistance to kanamycin (aphA-3), erythromycin (ermAM) and tetracycline (tetM). The 25.3 kb element is self-transferable to various Gram-positive bacterial genera where it transposes. Tn1545 was cloned in its entirety in the recombination deficient Escherichia coli HB101 where it was unstable. The three resistance genes aphA-3, ermAM and tetM were expressed but were not transferable to other E. coli cells. Tn1545 transposed from the hybrid plasmid to multiple sites of the chromosome of its new host. The element re-transposed, at a frequency of 5 X 10(-9), from the chromosome to various sites of a conjugative plasmid where it could be lost by apparently clean excision. The element transformed and transposed to the chromosome of Bacillus subtilis. The properties of the conjugative shuttle transposon Tn1545 may account for the recent emergence of genes from Gram positive bacteria in Gram-negative organisms. PMID- 3035334 TI - Molecular cloning and DNA homology of plasmid-mediated beta-lactamase genes. AB - Molecular cloning of DNA fragments between 1.5 and 8 kb from BamHI, EcoRI, HindIII, SalI, or Sau3A digests permitted the isolation of structural genes coding for TEM-1, ROB-1, OXA-1, OXA-3, OXA-4, OXA-5, PSE-1, PSE-2, PSE-3, PSE-4, CARB-3, CARB-4, AER-1, and LCR-1 beta-lactamases. Ampicillin-resistant clones were selected and it was confirmed that they contained the respective beta lactamase genes by isoelectric focusing. Detailed physical maps of 14 different recombinant plasmids were constructed using 8 restriction endonucleases. Plasmid deletions and lacZ fusions were used to localize the beta-lactamase structural genes. DNA probes were constructed for the TEM-1, ROB-1, OXA-1, and PSE-1 genes. Under conditions of high stringency, hybridization was observed between the genes for TEM-1 and TEM-2 or TLE-1, OXA-1 and OXA-4, and PSE-1 and PSE-4 or CARB-3, while the ROB-1 gene probe showed no cross-hybridization. Such bla gene probes should facilitate studies of beta-lactamase molecular epidemiology. PMID- 3035336 TI - Identification and mapping of regions that confer plasmid functions and of sites for excisive recombination of plasmid pMMC7105. AB - Strain PP808 of Pseudomonas syringae pv. phaseolicola contains pEXC8080 (34.6 kb), the smallest of several plasmids that originated by partial excision of the cryptic plasmid, pMMC7105 (150 kb), from the host chromosome. This excision plasmid is derived entirely of sequences from pMMC7105 and contains a 24 kb region referred to as common DNA, which is present in each of the other excision plasmids. A six enzyme restriction endonuclease map was constructed of pEXC8080. The replication region was mapped by identifying small restriction fragments that conferred replication properties to pMB1 plasmids that otherwise fail to replicate in Pseudomonas. This region is located within the common DNA and is 0.8 3.8 kb in size. Sequences from pEXC8080 failed to stabilize pMB1 derivatives in Pseudomonas in the absence of antibiotic selection, but stability functions were mapped to a region of pMMC7105 that presumably remains integrated in the chromosome of strain PP808. An incompatibility region was mapped to a 7.3 kb region on pEXC8080 that is closely linked to, but not included within, the replication region. The recombination site was mapped to a 1.2 kb region of the fusion fragment that was formed upon excision of pEXC8080. RS-I, a repetitive sequence found on pMMC7105 was present in the fusion fragment at the site of recombination. RS-I was also mapped to BamHI fragments that recombined upon excision of pEXC8080 and suggest that it provides sites for homologous recombination. PMID- 3035337 TI - Structure and expression of the overlapping ND4L and ND5 genes of Neurospora crassa mitochondria. AB - Genes homologous to the mammalian mitochondrial NADH dehydrogenase subunit genes ND4L and ND5 were identified in the mitochondrial genome of the filamentous fungus Neurospora crassa, and the structure and expression of these genes was examined. The ND4L gene (interrupted by one intervening sequence) potentially encodes an 89 residue long hydrophobic protein that shares about 26% homology (or 41% homology if conservative amino acid substitutions are allowed) with the analogous human mitochondrial protein. The ND5 gene (which contains two introns) encodes a 715 residue polypeptide that shares 23% homology with the human analogue; a 300 amino acid long region is highly conserved (50% homology) in the two ND5 proteins. The stop codon of the ND4L gene overlaps the initiation codon of the downstream ND5 gene, and the two genes are cotranscribed and probably cotranslated. A presumed mature dicistronic (ND4L plus ND5) RNA was detected. The postulated mRNA (about 3.2 kb) contains 5' and 3' non-coding regions of about 86 and 730 nucleotides, respectively; this species is generated from very large precursor RNAs by a complex processing pathway. The ND4L and ND5 introns are all stable after their excision from the precursor species. PMID- 3035338 TI - A cloned DNA fragment from bacteriophage P1 enhances IS2 insertion. AB - A 1.75 kb DNA segment of the bacteriophage P1 genome is known to serve as a preferred target for IS2 insertions. The presence of this fragment in a plasmid expressing the galK gene dramatically increases the proportion of IS2 insertions among spontaneous galK- mutants. Subfragments from two different parts of the 1.75 kb segment independently stimulate IS2 insertion, while another subfragment does not. In the plasmids studied IS2 elements not only insert into the cloned P1 fragment but also into parts of the galK gene with similar probability and mostly in one orientation. Many insertion sites are unique but several specific sites within the preferred target are repeatedly used for IS2 integration. The experimental data are compatible with a proposed cooperative mechanism, according to which more than one attracting sequence on the same plasmid might significantly enhance the probability of a particular target region to attract IS2. PMID- 3035339 TI - Cloning of seven differently complementing DNA fragments with chl functions from Escherichia coli K12. AB - Seven genomic libraries of chromosomal Escherichia coli K12 wild-type DNA were constructed in plasmid vectors. These were used to transform chl insertion mutants. Selection for growth on nitrate under anaerobic conditions yielded four plasmids which complemented mutants of the chlA, B, E and G types. The chromosomal fragments were mapped with restriction enzymes and subcloned. Three complementation groups were observed among the chlA mutants and two among the chlE mutants. The established complementation groups plus mutants of the chlD type represent eight distinct functions, which are all believed to be required for the molybdenum cofactor activity in the reduction of nitrate to nitrite by E. coli. PMID- 3035340 TI - Cloning and expression in Escherichia coli of the homoserine kinase (thrB) gene from Brevibacterium lactofermentum. AB - Five DNA fragments carrying the thrB gene (homoserine kinase E.C. 2.7.1.39) of Brevibacterium lactofermentum were cloned by complementation of Escherichia coli thrB mutants using pBR322 as vector. All the cloned fragments contained a common 3.1 kb DNA sequence. The cloned fragments hybridized among themselves and with a 9 kb BamHI fragment of the chromosomal DNA of B. lactofermentum but not with the DNA of E. coli. None of the cloned fragments were able to complement thrA and thrC mutations of E. coli. Plasmids pULTH2, pULTH8 and pULTH11 had the cloned DNA fragments in the same orientation and were very stable. On the contrary, plasmid pULTH18 was very unstable and showed the DNA inserted in the opposite direction. E. coli minicells transformed with plasmids pULTH8 or pULTH11 (both carrying the common 3.1 kb fragment) synthesize a protein with an Mr of 30,000 that is similar in size to the homoserine kinase of E. coli. PMID- 3035341 TI - Regulation of the ban gene containing operon of prophage P1. AB - A physical map of the ban gene of P1 and sites relevant to its regulation has been deduced from cloning of the appropriate regions of P1 wild-type and of P1 ban regulatory mutants. The cloning required the presence of P1 repressor in the cell confirming the existence of a repressible ban operon (Austin et al. 1978). Evidence for additional member(s) of that operon is presented. Of particular interest for understanding the regulation of ban are the relative positions of a binding site for the P1 repressor and of the regulatory mutations bac and crr that render ban expression constitutive. The results reveal a repressible operon like structure of about 4 kb within the P1 EcoRI-3 fragment that comprises a c1 repressor binding site/bac - additional gene(s) - crr/ban in the clockwise direction of the circular map of P1. PMID- 3035342 TI - Comparison of the organisation of the genomes of phenotypically diverse plasmids of incompatibility group P: members of the IncP beta sub-group are closely related. AB - Comparison of physical maps of the broad host range plasmids R751, R906 and R772, belonging to the IncP beta sub-group of the Escherichia coli incompatibility group P, reveals two large regions of similarity, separated by dissimilar regions which contain the majority of the cleavage sites for restriction endonucleases with hexanucleotide recognition sites. Mapping of the regions of these plasmids which show homology to probes specific for genetically characterised segments of the distantly related IncP alpha plasmid RK2, involved in plasmid maintenance or conjugal transfer, reveals that all four plasmids share a similar genetic organisation. In each case the homologous plasmid backbone is interrupted by heterologous segments both between the essential replication loci oriV and trfA, and between the conjugal transfer regions tra1 and tra2, although in the case of R772 the segment of the backbone carrying the trfA and tra2 regions is inverted relative to that of the other plasmids. However, in the case of pJP4, shown to be a fourth member of the IncP beta sub-group, the backbone is interrupted only by a single large segment adjacent to the trfA region. Mapping of the regions of the four IncP beta plasmids which show homology to Tn501 and nucleotide sequence determination at the ends of the homologous regions reveals that R906, R772 and pJP4 share a common mercury resistance region. This region, which appears to have been inactivated in R772, was probably inserted into a common ancestor of these plasmids by the transposition of an element related to an ancestor of Tn501. R751 shows no trace of the mercury resistance region, but contains a short relict of Tn501, derived from an independent insertion event. PMID- 3035343 TI - Replication of the streptococcal plasmid pMV158 and derivatives in cell-free extracts of Escherichia coli. AB - pMV158 is a 5.4 kb broad host range multicopy plasmid specifying tetracycline resistance. This plasmid and two of its derivatives, pLS1 and pLS5, are stably maintained and express their genetic information in gram-positive and gram negative hosts. The in vitro replication of plasmid pMV158 and its derivatives was studied in extracts prepared from plasmid-free Escherichia coli cells and the replicative characteristics of the streptococcal plasmids were compared to those of the E. coli replicons, ColE1 and the mini-R1 derivative pKN182. The optimal replicative activity of the E. coli extracts was found at a cellular phase of growth that corresponded to 2 g wet weight of cells per litre. Maximal synthesis of streptococcal plasmid DNA occurred after 90 min of incubation and at a temperature of 30 degrees C. The optimal concentration of template DNA was 40 micrograms/ml. Higher plasmid DNA concentrations resulted in a decrease in the incorporation of dTMP, indicating that competition of specific replication factor(s) for functional plasmid origins may occur. In vitro replication of plasmid pMV158 and its derivatives required the host RNA polymerase and de novo protein synthesis. The final products of the streptococcal plasmid DNAs replicated in the E. coli in vitro system were monomeric supercoiled DNA forms that had completed at least one round of replication, although a set of putative replicative intermediates could also be found. The results suggest that a specific plasmid-encoded factor is needed for the replication of the streptococcal plasmids. PMID- 3035344 TI - Cloning and characterization of the immunity region of phage phi 80. AB - The immunity region of phage phi 80 has been localized. It codes for at least three proteins: a protein of 34 kDa which has the biological properties of the phage repressor, and two other proteins of 9 kDa and 18 kDa which are the first proteins on the rightward operon. These two proteins are negatively regulated by the 34 kDa protein at a divergent promoter site. By position analogy with phage lambda, but not by its biological activity, the 9 kDa protein could be the cro product. The 18 kDa protein is able to block totally UV induction of phage phi 80. PMID- 3035345 TI - A transcriptional terminator sequence in the prokaryotic transposable element IS1. AB - The prokaryotic transposable element IS1 is known to exert a strong polar effect upon integration into an operon. To elucidate this polar effect, we constructed a plasmid which has an IS1 integrated between the 5' half of the tet gene for tetracycline resistance and the cat structural gene for chloramphenicol resistance. The cat gene is expressed by the tet promoter and the presence of IS1 in orientation I, in which the IS1 transposase genes insA and insB are in the same orientation as the cat gene, reduced the cat expression. By introducing deletions or insertions within the IS1 sequence, we were able to map a rho dependent terminator TIS1A between the insA and insB genes. Translational interruption between these ins genes is important for TIS1A to be an active terminator. PMID- 3035346 TI - Isolation and characterization of the regulatory HEX2 gene necessary for glucose repression in yeast. AB - The HEX2 gene which is necessary for glucose repression and is involved in the regulation of hexokinase PII synthesis and maltose uptake, has been cloned by complementation of a hex2 mutant, and selection for restored growth on maltose. Glucose repression in the transformants was like that in the wild type. The HEX2 gene was localized within a 2.15 kb fragment. The restriction map was confirmed by Southern hybridization of genomic DNA. Based on 30 tetrads, the linkage between HEX2 and TRP1 was determined as 10 cM. Plasmid integration directed to the genomic site of the cloned gene also gave a similar linkage distance between the amino acid auxotroph plasmid marker and genomic TRP1. Gene disruption of HEX2 yielded nonrepressible transformants with elevated hexokinase PII activity showing inhibition by maltose; this provides clear evidence that the HEX2 gene has been isolated. PMID- 3035347 TI - Replication of mini-F plasmid in vitro promoted by purified E protein. AB - An in vitro system for replication of mini-F plasmid DNA was constructed. This system consists of an ammonium sulfate fraction II (Fuller et al. 1981) from Escherichia coli extract, exogenously added purified E protein encoded by mini-F plasmid, and mini-F DNA in a closed circular form. Experiments with this system showed that the 217 bp DNA region which contains the A + T rich cluster and the four 19 bp direct repeats responsible for incB incompatibility is essential for mini-F DNA replication. PMID- 3035348 TI - Comparison of stationary and roller cultures for the isolation of herpesviruses affecting livestock. AB - The effect of rotating cultures on the isolation and cultivation of pseudorabies, infectious bovine rhinotracheitis and equine rhinopneumonitis viruses was studied. Serial ten-fold dilutions of laboratory strains of these viruses and field strains of pseudorabies virus were inoculated in two each of roller and stationary tubes of appropriate cell cultures. All tubes were examined daily for the appearance of cytopathic effects. Roller cultures were found to slightly enhance the speed and extent of cytopathic effect produced by these viruses. We conclude that it may be worthwhile to utilize roller cultures for isolation of animal viruses, especially for the recovery of low-titered virus, as is often the case with clinical specimens. PMID- 3035349 TI - The efficacy of biological response modifiers against murine cytomegalovirus infection in normal and immunodeficient mice. AB - The host-mediated antiviral effect of two biological response modifiers (BRM), OK 432 and PS-K, against murine cytomegalovirus (MCMV) was evaluated in normal and immunologically deficient mice of the same litters. In normal littermate mice, BALB/c (nu/+) or C57BL/6 (bg/+), the BRM-induced resistance against MCMV infection was evidenced by increase in fifty percent lethal doses, decrease in titers of viruses replicated in the target organs and augmentation of natural killer (NK) cell activity of the spleen cells. In T cell-deficient, athymic nude mice, BALB/c (nu/nu), the protective effect was manifested by prolongation of the survival, decrease in the virus titers, and increase in the NK-cell activity, but without decrease in mortality. In NK cell-deficient, beige mutant mice, C57BL/6 (bg/bg), the BRM-induced protection was nullified or minimized, and there was little difference in those parameters between BRM-treated and untreated mice. However, with higher doses of OK-432, but not PS-K, or with sublethal doses of MCMV, the NK cell activity was slightly augmented in the beige mutant mice. Thus both NK cell and T cell activity are essential for mice to overcome acute MCMV infection and it is likely that the protective effect of BRM manifests itself fully, at least in immunologically intact mice. PMID- 3035350 TI - Leukaemogenesis: a postulated mechanism involving tyrosine protein kinase and DNA topoisomerase. AB - The weight of available evidence suggests that leukaemogenesis is a multi-step, complex process. Since there are different types of leukaemia, it seems likely that a variety of distinct molecular mechanisms are involved, arising from different combinations of genetic defects. It is not therefore surprising that studies at genetic, karyotypic, biochemical, cellular and clinical levels all reveal considerable heterogeneity of abnormalities. An attempt to define associations between abnormalities at these different levels in the myeloid leukaemias, led to a new hypothesis which provides a link between activation of the tyrosine kinase group of oncogenes and two components of multi-step leukaemogenesis: reduced differentiation and the tendency for acquisition of further genetic changes. PMID- 3035351 TI - A role for glucocorticoids in the polyphosphoinositide second messenger system. AB - Glucocorticoids have been shown to be involved in numerous secretory and activation processes which are known to be mediated by the polyphosphoinositide second messenger system. A connection between glucocorticoids and the polyphosphoinositide system has not been made because of the marked temporal differences in their effects and the fact that most of the known effects of glucocorticoids involve transcription and/or protein synthesis. An attempt is made to to rationalize these apparent incongruities. The recently reported stimulation of glucose transport by kinase C suggests an experimental system to investigate glucocorticoid effects on the polyphosphoinositide system. PMID- 3035352 TI - Epstein-Barr virus as an etiological agent in primary Sjogren's syndrome. AB - It is proposed that the initiating event in primary Sjogren's syndrome is infection with Epstein-Barr virus (EBV), and that the autoimmune exocrinopathy that progresses to keratoconjunctivitis sicca and xerostomia is a sequel to this. Our hypothesis is based on the findings that the antinuclear antibody, anti-La(SS B), is a marker of primary Sjogren's syndrome, and that anti-La reacts with a ribonucleoprotein, the La autoantigen, to which bind not only all cellular RNAs transcribed by RNA polymerase III, but also the viral RNAs, EBER 1 and EBER 2, encoded by the Epstein-Barr virus (EBV). It is proposed that during EBV infection, there are multiple copies of the EBERs available to bind to the La ribonucleoprotein and when infection occurs in subjects who have an impaired T cell-mediated response to EBV, and who are genetically predisposed to autoimmunity, there is loss of immunological tolerance to La with production of anti-La (SS-B). Thus the inflammatory process in exocrine glands which culminates in the sicca syndrome is due to the combined effects of chronic EBV infection and autoimmunity. Two patients are briefly described in whom primary Sjogren's syndrome appeared to be a direct consequence of EBV infection. The hypothesis engages the question of the respective roles of virus infection, specific immunodeficiency to virus, immunogenetic constitutive influences and an autoimmune response to a ribonucleoprotein antigen in the genesis of a particular organ-specific inflammatory reaction. PMID- 3035353 TI - Low prevalences of coronary heart disease (CHD), psoriasis, asthma and rheumatoid arthritis in Eskimos: are they caused by high dietary intake of eicosapentaenoic acid (EPA), a genetic variation of essential fatty acid (EFA) metabolism or a combination of both? AB - The low prevalences of CHD, psoriasis, asthma and rheumatoid arthritis in Eskimos have been attribute to the high dietary intake of EPA from fish and marine mammals. However, even on a Western diet, Eskimos have plasma arachidonic acid (AA) levels far below those seen in Europeans while dihomogammalinolenic acid (DGLA) levels are higher in Eskimos. These low AA and high DGLA levels seem to be due to a genetic abnormality in EFA desaturation since they are found even when EPA intakes are low. Since AA is known to be important in the pathogenesis of CHD, asthma, psoriasis and arthritis, while DGLA has properties which make it of likely therapeutic value in these conditions, the genetically high DGLA and low AA are likely to be as important as dietary EPA in determining Eskimo disease patterns. PMID- 3035354 TI - Hodgkin's disease following Wilms' tumor: a case report. AB - A patient is described who developed Hodgkin's disease 16 years after treatment of Wilms' tumor with radiation and actinomycin D. The issues relating to Hodgkin's disease as a second malignancy are reviewed. The need for long-term surveillance of patients with malignant disease for late effects of the cancer and its treatment is emphasized. PMID- 3035355 TI - [New views on lipoprotein metabolism]. PMID- 3035356 TI - [Viral infections in female genital organs]. PMID- 3035357 TI - Pseudohypoparathyroidism. An update. PMID- 3035359 TI - Self-study training offered by CDC. PMID- 3035358 TI - Two sibs affected by Pendred's syndrome in a family with recurrent goiter. PMID- 3035360 TI - National Center for Health Statistics joins CDC. PMID- 3035361 TI - [Prosthetic reconstruction of the trachea and carina]. AB - We have performed prosthetic reconstruction of the trachea and carina in 12 patients since 1979. We used Neville's prosthesis in 7 patients and Katsura's prosthesis in 5 patients. Seven patients were operated on for lung cancer, 2 patients for adenoid cystic carcinoma of the trachea and the others for large cell carcinoma of the trachea, thyroid cancer and tuberculous granuloma, respectively. Prosthetic reconstruction of the trachea was performed in 4 patients. Carinal resection was performed in 8 patients: With right sleeve pneumonectomy in 4 patients, with right upper lobectomy in 2 patients and only carinal resection in 2 patients. Prosthetic reconstruction after the carinal resection was performed using 3 straight types, 1 curved type and 4 bifurcated types. With regard to the complications of the prosthetic reconstruction, dehiscence at the anastomotic site was seen in 5 patients, granulation in 4 patients, empyema in 3 patients, massive hemorrhage in 2 patients and migration of the prosthesis in 1 patient. Five patients survived more than 1 year. The longest survival time was 43 months. To prevent complications of the prosthetic reconstruction, we improved the anastomotic method, reinforced the anastomotic site with Marlex mesh and protected the surrounding vessels with Lyodura. PMID- 3035362 TI - Glucocorticoid induction of beta-adrenergic receptors in the DDT1 MF-2 smooth muscle cell line involves synthesis of new receptor. AB - We have shown that glucocorticoids induce the appearance of beta 2-adrenergic receptors in membranes of the ductus deferens smooth muscle cell line (DDT1 MF 2). A concomitant increase in isoproterenol stimulated adenylate cyclase activity in the absence of exogenously applied GTP was observed as was a significantly increased (p less than 0.05) sensitivity of the adenylate cyclase system to exogenously applied GTP. However, no significant difference in the maximal velocity of adenylate cyclase between control and steroid treatment was measurable in the presence of sodium fluoride. Induction of beta 2-adrenergic receptors in DDT1 MF-2 cells is correlated with the presence of steroid receptors (androgen and glucocorticoid) in the cells since estrogens and progesterones had no effect on receptor levels. Finally, utilizing dense amino acid labeling of cells to measure old versus newly synthesized receptor sites by a density shift method, we have documented that glucocorticoid induction of beta 2-adrenergic receptors involves synthesis of new receptor protein. PMID- 3035365 TI - [New blood pressure lowering drugs in childhood]. AB - The treatment by antihypertensive agents has recently been improved by the introduction of angiotensin converting-enzyme inhibitors, and of calcium antagonists, which have been applied successfully in children with acute and chronic hypertension. The converting enzyme inhibitor captopril is used increasingly for the treatment of severe forms of persistent hypertension in children. Side effects from this drug can largely be avoided by using low daily doses. The calcium-antagonist nifedipine can now be regarded as the drug of choice in the treatment of acute elevation of blood pressure in infants and children. For this indication it appears to be superior to other drugs because of its prompt and safe efficacy after sublingual administration. PMID- 3035364 TI - Activation of phosphoprotein phosphatases by growth hormone sequences with insulin-like activity. AB - The N-terminal part sequences of pituitary growth hormone, N alpha-acetyl-hGH 7 13 and hGH 6-13, promoted conversion of glycogen synthase b to glycogen synthase a in skeletal muscle and adipose tissue when injected intravenously. The peptides also caused conversion of phosphorylase a to phosphorylase b in liver and adipose tissue, but not in muscle, where the peptides antagonised activation of phosphorylase. Synthase phosphatase activity in muscle and phosphorylase phosphatase activity in liver increased after injection of peptide, with time courses of change similar to those seen for muscle synthase and liver phosphorylase activities. Injection of peptide also decreased both the cyclic AMP dependent and independent synthase kinase activities in muscle. These results show that the insulin-like activities of these peptides on glycogen synthase and phosphorylase involve both increases in protein phosphatase activities and inhibition of protein kinase activities. These results are discussed in relation to the insulin-like activities of growth hormone. PMID- 3035366 TI - [Results of virologic studies in 620 children with abacterial meningitis over a 15-year period (1971-1985)]. AB - Cases of non-bacterial meningitis being hospitalized at the Children's Hospital of the University of Bochum in the years 1971 to 1985 were analysed retrospectively according to etiological and epidemiological aspects. Altogether 620 non-bacterial cases of meningitis were included in this study. For 378 (60.9%) children the infecting agent could be identified while in 242 cases (39.1%) the etiology remained open. In 237 cases enterovirus had been the causative agent. The average age of the patients was 6.5 years, boys were more frequently involved than girls (65% versus 35%). The majority of enterovirus infections occurred during the summer months July to September whereas (mumps) parotitis infections were equally distributed over the whole year. Yearly increases in certain enterovirus types were correlated to an increase in ECHO 11 infections in 1982, Coxsackie B5-infection in 1983 and 84 and ECHO 7-infection in 1985 based on data from the whole Federal Republic of Germany. However the majority of enterovirus-caused meningitis occurred during local endemics. PMID- 3035367 TI - The pathology of thymic neoplasia. PMID- 3035369 TI - Relative refractory period: a measure to detect early neuropathy in alcoholics. AB - The absolute (ARP) and relative refractory period (RRP) of the median sensory nerve was determined in 26 control subjects and 24 alcoholics, nine of whom had symptoms of peripheral neuropathy. Recovery of latency to normal in response to the second stimulus was used to define RRP. A true RRP was calculated by subtracting ARP from measured RRP. Mean ARP for all subjects ranged from 0.75 to 0.80 msec; normal = 0.8 +/- 0.2 msec. The true RRP of control subjects was 2.1 +/ 0.5 msec, and for all alcoholic subjects it was 3.1 +/- 0.5 msec. True RRP for the nine symptomatic alcoholic subjects was 3.6 +/- 0.5 msec and 2.9 +/- 0.4 msec for those who were asymptomatic. Symptomatic and asymptomatic alcoholics differed significantly from one another, as well as from control subjects (P less than 0.001). Routine nerve conduction studies were normal in asymptomatic subjects. Three out of nine symptomatic alcoholics had increases in distal median motor or sensory latency, and three had slight slowing of median nerve conduction velocity. True RRP is more sensitive than routine measures of nerve conduction in the detection of axonal disorders influencing nerve conduction. PMID- 3035363 TI - Effects of ATP and cyclic AMP on the in vitro assembly and stability of mammalian brain microtubules. AB - The relevance of protein phosphorylation, transphosphorylation and binding phenomena in the kinetics of the ATP-induced assembly of cycle-purified microtubule protein from mammalian brain were studied. ATP was able to induce the polymerization of microtubules of normal appearance. However, the assembled structures, were unstable and microtubules depolymerized after achievement of a transitory maximum. Cyclic AMP reduced the amplitude of the polymerization maximum in a concentration-dependent manner, correlating with the stimulation of the endogenous phosphorylation reaction. When microtubule assembly was induced by GTP, in the presence of various concentrations of ATP, the slope of the depolymerization phase was found to depend on the concentration of ATP. Fluoride ion inhibited the endogenous phosphorylation reaction and reduced the disassembly rate, in a concentration-dependent manner. Evidence is also presented indicating that ATP did not bind to phosphocellulose-purified tubulin. These results further contribute to indicate that ATP and cyclic AMP, acting coordinately to control the phosphorylation extent of microtubule proteins are important factors to determine microtubule stability within the cell. Some implications of this mechanism for the regulation by cAMP of the initiation of DNA synthesis and mitosis are considered. PMID- 3035368 TI - Overnutrition in the diabetic patient. PMID- 3035370 TI - Studies on the role of macrophages in experimental candidosis in mice. PMID- 3035371 TI - Fungistatic and fungicidal effects of amphotericin B, ketoconazole and fluconazole (UK 49,858) against histoplasma capsulatum in vitro and in vivo. PMID- 3035372 TI - Colchicine myopathy and neuropathy. AB - Although colchicine has been used for centuries, its neuromuscular toxicity in humans is largely unrecognized. In this report we describe a characteristic syndrome of myopathy and neuropathy and present 12 new cases of the condition. Colchicine myopathy may occur in patients with gout who take customary doses of the drug but who have elevated plasma drug levels because of altered renal function. It usually presents with proximal weakness and always presents with elevation of serum creatine kinase; both features remit within three to four weeks after the drug is discontinued. The accompanying axonal polyneuropathy is mild and resolves slowly. Electromyography of proximal muscles shows a myopathy that is marked by abnormal spontaneous activity. Because of these features, colchicine myoneuropathy is usually misdiagnosed initially, either as probable polymyositis or as uremic neuropathy. The myopathy is vacuolar, marked by accumulation of lysosomes and autophagic vacuoles unrelated to necrosis or to the mild denervation in distal muscles. The morphologic changes in muscle suggest that the pathogenesis involves disruption of a microtubule-dependent cytoskeletal network that interacts with lysosomes. Correct diagnosis may save patients with this disorder from inappropriate therapy. PMID- 3035373 TI - Reduced binding of [3H]1,25-dihydroxyvitamin D3 in the parathyroid glands of patients with renal failure. AB - This study examined the hypothesis that altered binding of 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3) to parathyroid receptors might be involved in the pathogenesis of secondary hyperparathyroidism associated with chronic renal failure. The binding of [3H]1,25-(OH)2D3 to hyperplastic parathyroid glands obtained from seven patients with chronic renal failure was measured. These values were compared with those for binding to hyperplastic parathyroid tissue obtained from six patients who had received renal transplants and for binding to parathyroid adenomas removed from five patients who had primary hyperparathyroidism. We found that Nmax (an estimate of the concentration of 1,25-(OH)2D3 receptors) was reduced (42 +/- 15 fmol per milligram of protein) in patients with chronic renal failure as compared with patients with transplanted kidneys (78 +/- 24 fmol per milligram of protein) and patients with primary hyperparathyroidism (114 +/- 30). Nmax correlated inversely with the severity of renal dysfunction, the serum level of phosphorus, and the logarithm of the serum level of immunoreactive parathyroid hormone. These observations suggest that 1,25-(OH)2D3 binding by parathyroid tissue is reduced in chronic renal failure. This may contribute to the pathogenesis of secondary hyperparathyroidism by reducing the inhibition by 1,25 (OH)2D of parathyroid hormone secretion. The low serum levels of 1,25-(OH)2D in chronic renal failure may accentuate this effect. PMID- 3035374 TI - Development of cytomegalovirus (CMV) retinitis in a patient with AIDS during ganciclovir therapy for CMV colitis. PMID- 3035376 TI - Obtaining access to data from government-sponsored medical research. PMID- 3035375 TI - Selective hypoaldosteronism due to an endogenous impairment in angiotensin II production. PMID- 3035377 TI - The third component of complement (C3) is responsible for the intracellular survival of Leishmania major. AB - Leishmania are obligate intracellular parasites of mononuclear phagocytes. We and others have shown that the promastigote form of all species of leishmania activates complement from non-immune serum and that this activation can result in parasite lysis. This work, as well as earlier in vivo studies, suggested that complement is an important component of host defence against leishmaniasis. We now present evidence that parasite complement fixation, in addition to increasing parasite phagocytosis, is required for the intracellular survival of leishmania in macrophages. We specifically show a strong correlation between parasite C3 fixation and intracellular survival. We attribute this survival, in part, to a decrease in the magnitude of the macrophage respiratory burst which is triggered by complement-coated, as opposed to uncoated, parasites. PMID- 3035378 TI - A DNA sequence specific for forest form Onchocerca volvulus. AB - Onchocerciasis, or river blindness, is caused by infection with Onchocerca volvulus, a filarial parasite which infects about 40 million people in Africa and Latin America. Epidemiological, clinical, entomological and serological studies of African onchocerciasis led to the hypothesis that Onchocerca volvulus exists in different forms in the forest and savannah. It is uncertain if these differences are due to genetic differences within O. volvulus itself, or to epigenetic factors, such as differences in the host populations. To date no basic biochemical differences between the forest and savannah populations of O. volvulus has been found, although isoenzyme studies have shown that differences in allele frequency between forest and savannah populations exist. Here we describe the isolation of a DNA sequence that seems to be specific for the forest form of O. volvulus, the first indication of a basic genetic difference between the savannah and forest forms. PMID- 3035379 TI - AIDS research: human and monkey virus puzzles. PMID- 3035380 TI - Myristylation of picornavirus capsid protein VP4 and its structural significance. AB - We have obtained evidence that poliovirus and other picornavirus particles are specifically modified by having myristic acid covalently bound to a capsid protein. The electron density map of poliovirus confirms the position of the myristate molecule and defines its location in the virus particle. Analogies with other myristylated proteins suggest that the myristate moiety in picornaviruses may be involved in capsid assembly or in the entry of virus into cells. PMID- 3035381 TI - NMDA receptors in the visual cortex of young kittens are more effective than those of adult cats. AB - Acidic amino acids, such as glutamate and aspartate, are thought to be excitatory transmitters in the cerebral neocortex and hippocampus. Receptors for these amino acids can be classified into at least three types on the basis of their agonists. Quisqualate-preferring receptors and kainate-preferring receptors are implicated in the mediation of synaptic transmission in many regions including the hippocampus and visual cortex, whereas N-methyl-D-aspartate (NMDA)-preferring receptors are thought to be involved in modulating synaptic efficacy, for example in longterm potentiation, a form of synaptic plasticity in the hippocampus. In the visual cortex of the cat and monkey, it is well established that synaptic plasticity, estimated by susceptibility of binocular responsiveness of cortical neurons to monocular visual deprivation, disappears after the 'critical' period of postnatal development. Here we report that during the critical period in young kittens, a selective NMDA-receptor antagonist blocks visual responses of cortical neurons much more effectively than it does in the adult cat. This suggests that NMDA receptors may be involved in establishing synaptic plasticity in the kitten visual cortex. PMID- 3035382 TI - Different recombination site specificity of two developmentally regulated genome rearrangements. AB - In the absence of a combined nitrogen source, such as ammonia, approximately every tenth vegetative cell along filaments of the cyanobacterium Anabaena develops into a heterocyst, a terminally differentiated cell that is morphologically and biochemically specialized for nitrogen fixation. At least two specific DNA rearrangements involving the nitrogen-fixation (nif) genes occur during heterocyst differentiation, one within the nifD gene and the other near the nifS gene. The two rearrangements have several properties in common. Both occur quantitatively in all heterocyst genomes, both occur at approximately the same developmental time, late in the process of heterocyst differentiation, and both result from site-specific recombination between short repeated DNA sequences. We report here the nucleotide sequences found at the site of recombination near the nifS gene. These sequences differ from those found previously for the nifD rearrangement, suggesting that the two rearrangements are catalysed by different enzymes and may be regulated independently. We also show that the nifS gene is transcribed only from rearranged genomes. PMID- 3035384 TI - Stereoselective inhibition of calmodulin-dependent cAMP phosphodiesterase from bovine heart by (+)- and (-)-nimodipine. AB - The inhibitory effects of racemic (+/-)-nimodipine and of optically pure (+)- and (-)-nimodipine on the basal and calmodulin-dependent activity of a cAMP phosphodiesterase from bovine heart were investigated. The inhibition by (+/-) nimodipine could not be overcome by an excess of calmodulin. However, increase of the cAMP concentration in the assay from 2 X 10(-4) mol/l to 2 X 10(-2) mol/l caused a shift of the IC50 for the inhibition by (+/-)-nimodipine from 2.8 X 10( 6) mol/l to 6 X 10(-5) mol/l. Dixon-plot analysis revealed an inhibitory constant of Ki = 2.3 mumol/l. Experiments with the two enantiomers showed that (+) nimodipine is by about one order of magnitude more potent than (-)-nimodipine. This contrasts with the stereoselectivity of the Ca2+ channel inhibitory activity on isolated rings of the rabbit basilar artery where (-)-nimodipine is more effective than (+)-nimodipine in relaxing the smooth muscle contracted by K+ depolarisation. It is concluded that cAMP phosphodiesterase may be an intracellular target for nimodipine and its inhibition may contribute to the pharmacological activity of this 1,4-dihydropyridine. PMID- 3035383 TI - Inhibition of dopamine-sensitive adenylate cyclase by opioids: possible involvement of physically associated mu- and delta-opioid receptors. AB - D-1 dopamine receptor-stimulated cyclic AMP efflux from rat neostriatal slices (induced by 30 microM dopamine + 10 microM (-)sulpiride) was concentration dependently reduced by morphine, [D-Ala-D-Leu]-enkephalin (DADLE), [D-Pen-D Pen]enkephalin (DPDPE) and bremazocine. Naloxone (0.1 microM) selectively antagonized the inhibitory effect of (a submaximally effective concentration of) morphine, whereas ICI 174864 (0.75 microM) completely blocked the inhibitory effects of DADLE, DPDPE and bremazocine without affecting that of morphine, indicating a role of mu- as well as delta-opioid receptors. Upon simultaneous activation of D-1 dopamine receptors and delta-opioid receptors the (mu-receptor mediated) inhibitory effect of morphine was abolished, while it was not changed following simultaneous activation of D-1 and (inhibitory) D-2 dopamine receptors. Cyclic AMP efflux induced by isoprenaline or adenosine was not affected by the opioids and that induced by vasoactive intestinal peptide (VIP) was inhibited by morphine and DADLE only. In the latter case naloxone, but not ICI 174864, antagonized the inhibitory effects. These data show that D-1 dopamine receptor stimulated adenylate cyclase activity in rat neostriatum, but not that stimulated through other receptors, is inhibited by two pharmacologically distinct opioid receptor subtypes. It is speculated that these mu- and delta-opioid receptors share a common inhibitory guanine nucleotide binding protein and may represent closely associated recognition sites of a functional opioid receptor complex. PMID- 3035385 TI - Direct activation by okadaic acid of the contractile elements in the smooth muscle of guinea-pig taenia coli. AB - Okadaic acid isolated from black sponge (Halichondria okadai), at the concentration of 10 mumol/l, caused contraction in saponin-treated skinned smooth muscle of guinea-pig taenia coli in the absence of Ca2+. In the presence of low concentration (0.3 mumol/l) of Ca2+, okadaic acid induced a greater contraction than in the absence of Ca2+. Okadaic acid potentiated the contractions induced by Ca2+ and pCa2+-tension curve was shifted to the left as well as upward by 1 mumol/l okadaic acid. Native actomyosin preparation (myosin B) containing calmodulinmyosin light chain kinase system and phosphatase was obtained from taenia coli. Okadaic acid (10 mumol/l) increased the actomyosin Mg2+-ATPase activity in the presence or absence of Ca2+. Okadaic acid (1-100 mumol/l) had no effect on calmodulin activity as monitored by Ca2+-calmodulin activated cyclic nucleotide phosphodiesterase activity and the (Ca2+ + Mg2+)-ATPase activity or erythrocyte membranes. These results suggest that okadaic acid directly activates contractile elements of smooth muscle. PMID- 3035386 TI - [Incidence of granular cell myoblastoma, 1980-1985]. PMID- 3035387 TI - [Transient inhibition of adrenal cortex function following induction of anesthesia with etomidate]. PMID- 3035388 TI - "The use of Rh-immune globulin". PMID- 3035389 TI - [Encephalitis with isolated pupillary disorder and ganglioradiculo-neuropathy with ascending degeneration of the posterior column in bronchial cancer]. PMID- 3035390 TI - [Polyneuropathy and necrotizing myopathy following desensitization therapy]. PMID- 3035391 TI - Drug resistance in medical oncology. PMID- 3035392 TI - Plasma levels of main granulocyte components in patients dialyzed with polycarbonate and cuprophan membranes. AB - Plasma levels of granulocyte lactoferrin, granulocyte myeloperoxidase and granulocyte elastase in complex with alpha 1-proteinase inhibitor (E-alpha 1PI) were investigated in regular hemodialysis patients dialyzed with hollow-fiber dialyzers made from polycarbonate (FD 100) or cuprophan (GFS 120 H). Plasma levels of all these main granulocyte components increased significantly during hemodialysis. E-alpha 1PI levels were significantly higher in patients dialyzed with the polycarbonate compared with the cuprophan membrane, whereas the increases of myeloperoxidase and lactoferrin were not different for the two dialyzers. On the other hand, plasma C3a levels were higher in patients dialyzed with the cuprophan compared with the polycarbonate dialyzer. Therefore, granulocyte activation during hemodialysis does not necessarily need complement activation. PMID- 3035393 TI - Reversible acute renal insufficiency with combination of enalapril and diuretics in a patient with a single renal-artery stenosis. PMID- 3035394 TI - Serum angiotensin-converting enzyme during haemodialysis: effect of membrane biocompatibility. PMID- 3035395 TI - [Regulation of energy metabolism in the cerebral cortex by VIP (vasoactive intestinal peptide) neurons: an example of peptidergic neural transmission]. PMID- 3035396 TI - [Naloxone-reversible analgesia as a result of stimulation of the habenula in the rat]. AB - Electrical stimulation of rat habenular complex induces analgesia, evaluated by the tail-flick test, dependent on intensity of stimulation with a long post effect, that is reversible by naloxone and without behavior effects at less that 400 mA. Bilateral destruction of habenula fails to provoke hyperesthesia but causes more marked long-term tolerance effects than in controls. Anatomy suggests that the habenula activates an inhibitory descending system in the spinal cord with a probable relay in the dorsal raphe and involving an endogenous opioid dependent stage. PMID- 3035398 TI - [Cytochemical demonstration of an endogenous inhibitor of Na-K ATPase and its relationship to familial arterial hypertension]. AB - Acute volume expansion, an increase in sodium intake and a restraint on sodium excretion endow the plasma with the capacity to cause a natriuresis, to inhibit sodium transport and to stimulate vascular reactivity. One natriuretic substance, the atrial natriuretic peptide, has been identified. Cytochemical techniques can detect the presence of a Na-K ATPase inhibitor in the plasma of normal man and the rat, the concentration of which is controlled by salt intake. The substance responsible appears to originate in the hypothalamus. The plasma concentration of the cytochemically detectable Na-K ATPase inhibitor is substantially raised in the plasma of patients with essential hypertension, of the spontaneously hypertensive rat and of the Milan hypertensive rat. An hypothesis is put forward that links salt intake, a genetic renal lesion, the endogenous Na-K ATPase inhibitor, the atrial natriuretic peptide, and the substance responsible for vascular reactivity, with the rise in arterial pressure in hereditary forms of hypertension. PMID- 3035399 TI - Opioid receptor binding in rat spinal cord. AB - The interaction of various radioligands with spinal opioid receptors has been characterized under variable experimental conditions. Binding to mu, delta, and kappa sites was measured in all (cervical, thoracic, lumbar) segments. The apparent affinity constant (K) of [3H]Ethylketocyclazocine (EKC) was similar in Tris, 2.09 (+/- 1.06) X 10(8) M-1, and phosphate buffer, 2.16 (+/- 0.02) X 10(8) M-1, when its interaction with delta and mu sites was blocked. Without blocking ligands, EKC binding was resolved in two components: K1 = 1.01 (+/- 0.21) X 10(9) M-1 and K2 = 0.95 (+/- 0.61) X 10(7) M-1. Likewise, the binding of [D-Ala2, MePhe4, Gly(ol)5]enkephalin (DAGO) or [D-Ala2, D-Leu5]-enkephalin (DADLE) alone was represented by a 2-site model. By adjusting the radioligand and receptor concentration or by the addition of blocking ligands, binding was represented by a 1-site model for DAGO, K = 4.35 (+/- 1.41) X 10(8) M-1, and DADLE, K = 2.44 (+/ 0.08) X 10(8) M-1. PMID- 3035397 TI - [Development of renal function in the transplanted patient with renal hypertension treated with enalapril]. AB - We report our experience with a converting-enzyme inhibitor, enalapril, as antihypertensive agent in eighteen patients with hypertension after renal transplantation. Renal function was prospectively followed up. Six patients demonstrated an acute renal failure episode (defined by a 25% increase of serum creatinine during enalapril therapy). Renal failure was always reversible with interruption or dosage reduction of the drug. We recommend to start therapy with low dose and to closely monitor renal function. PMID- 3035400 TI - Brain cortical amino acids measured by intracerebral dialysis in portacaval shunted rats. AB - The extracellular amino acid content was measured in the parietal cortex in portacaval and sham operated rats, using the brain dialysis technique. The amino acid content of the perfusate was determined for 10 min before and during stimulation with potassium chloride. Basal levels of aspartate, glutamine, glycine, methionie, valine, phenylalanine and leucine were 2-to 6-fold higher in the PC-shunted as compared to the sham operated rats. For glutamate, taurine, and GABA no differences were observed between the two groups. After KCl stimulation the release of glutamate and GABA increased significantly in both groups. For GABA this rise was approximately twice as high in the PC-shunted rats (+300%, P less than 0.01) as in the sham operated rats (+150%, P less than 0.01 as compared to basal). In the sham operated, but not in the PC-shunted rats, methionine and valine levels rose significantly (+200%, P less than 0.05) and glutamine release decreased (-50%, P less than 0.05). These findings suggest that the brain metabolism of amino acids is altered after a portacaval shunt. This could in turn alter the neurotransmission and partly explain the low spontaneous motor activity seen in these animals. PMID- 3035401 TI - Differential maturation of mu and delta opioid receptors in the chick embryonic brain. AB - The developmental profiles of the binding of mu and delta opiate receptors agonists was investigated using the chick embryo brain. Binding of opioids was performed at embryonic days 5, 6, 15, 18, and 20 in the developing chick embryo brain. [3H]dihyromorphine was used as a mu ligand and with 5 X 10(-7) M levorphanol for non-specific binding, and [3H](D-Ala2-D-Leu5)-enkephalin was used as a delta with 5 X 10(-7) M (D-Ser-Gly-Phe-Leu-Thr)-enkephalin for non-specific binding. Crude membranes were prepared from whole brain at days 5, 6 and cerebral hemispheres at days 15, 18, and 20 of embryonic age. Both mu and delta opiate receptors were present during early embryogenesis and as early as day 5. Analysis of binding sites revealed high and low affinity mu sites during early embryogenesis but only one delta site. By 18 days of embryonic age, only one mu site remained. This developmental change is interpreted as a transitory state of the receptor to the adult mu pattern. The presence of only one delta site is constant throughout embryonic age; it is high during early embryogenesis reaching a lower level by 18 days. The presence of a dual binding site pattern for the mu receptor in early embryogenesis is implicated to have a functional significance in the pluripotential role of the endogenous opioids in early development. PMID- 3035403 TI - A method for the determination of the integrity of labeled GABA used in membrane receptor binding assay. AB - A simple method for the determination of the proportion of true GABA within labeled "GABA" used for membrane binding assay is presented. The method is intended for the assessment of the integrity of refrigerator (+4 degrees C) stored labeled neurotransmitter. Its application allows a precise determination of the binding parameters. PMID- 3035402 TI - Cerebral glycine content and phosphoserine phosphatase activity in hyperaminoacidemias. AB - Chronic hyperphenylalaninemia maintained with the aid of a suppressor of phenylalanine hydroxylase, alpha-methylphenylalanine, increases the glycine concentration and the phosphoserine phosphatase activity of the developing rat brain but not that of liver or kidney. Similar increases occur after daily injections with large doses of phenylalanine alone, while tyrosine, isoleucine, alanine, proline, and threonine, were without effect. Treatment with methionine, which increases the phosphoserine phosphatase activity of the brain and lowered that of liver and kidney, left the cerebral glycine level unchanged. When varying the degrees of gestational or early postnatal hyperphenylalaninemia, a significant linear correlation was found between the developing brains' phosphoserine phosphatase and glycine concentration. Observations on the uptake of injected glycine and its decline further indicate that coordinated rises in the brain's phosphoserine phosphatase and glycine content associated with experimental hyperphenylalaninemia denote a direct impact of phenylalanine on the intracellular pathway of glycine synthesis in immature animals. PMID- 3035404 TI - Effects of systematically administered lithium on tryptophan transport and exchange in plasma-membrane vesicles isolated from rat brain. AB - The effect of lithium on the sodium-dependent high-affinity system for tryptophan uptake was examined in plasma membrane vesicles derived from rat brain. We demonstrated that Na+ could be replaced by lithium in the external medium and the presence of lithium produced an increase in the Vmax of the tryptophan transport whereas it had no significant effect on the Km for the substrate. Plasma membrane vesicles derived from synaptosomes obtained from long-term lithium-treated rats are able to accumulate tryptophan to a greater extent than normal rats and maintain a more negative membrane potential than controls. Our data support the idea that the stimulation by lithium of the high-affinity uptake system for tryptophan by maintaining adequate membrane potentials across the membrane, could lead to the stabilization of serotonin production, as has been demonstrated in long term-lithium treatment. PMID- 3035406 TI - [Effect of ACTH therapy on IgG synthesis in cerebrospinal fluid spaces in patients with disseminated sclerosis]. AB - In 33 cases of multiple sclerosis treated with ACTH (2500 u during 40 days) IgG and albumins were determined in the serum and cerebrospinal fluid by Mancini's method. These determinations served for calculation of the IgG: albumin ration and IgG index. After the treatment a significant reduction was found of the serum level of IgG and IgG index, without significant changes in albumin concentration. The effect of ACTH therapy on IgG synthesis within the CSF spaces was not significant in the whole group. In the subgroup of patients with initially demonstrated IgG synthesis within the CSF spaces (before the treatment) a significant decrease was found of the IgG index and a non-significant decrease of the absolute IgG level in the CSF. A comparison of the results with our previous investigations shows that ACTH, despite its positive effect on multiple sclerosis exacerbations exerts a much weaker effect on the local IgG synthesis in the central nervous system than high doses of prednisone, despite an evident reduction of serum IgG level by ACTH. PMID- 3035407 TI - Effects of interleukin-1 on hormone release from normal rat pituitary cells in primary culture. AB - The present study was performed mainly to determine whether interleukin-1 (IL-1), a polypeptide produced by immunologically activated monocytes, plays a physiological role in the regulation of adrenocorticotropic hormone (ACTH) using primary monolayer cultures of rat anterior pituitary cells. Neither human IL-1 alpha nor IL-1 beta stimulated the ACTH release from normal pituitary cells in concentrations ranging from 0.01 to 10 nM. IL-1 beta caused a slight, but significant, increase in ACTH release at a concentration of 100 nM, while IL-1 alpha did not, even at the highest dose tested. IL-1 beta exhibited a synergistic action with corticotropin-releasing factor (CRF) in ACTH secretion at 10 and 100 nM of CRF, but the interaction was not striking. Both of the monokines failed to cause any change in the secretions of growth hormone, prolactin, follicle stimulating hormone and luteinizing hormone throughout concentrations ranging from 0.01 to 100 nM. The effects of possible sex-related differences and prolonged preincubation of cultured pituitary cells in serum-free medium prior to assay incubation were also tested, providing no significantly different findings. These results suggest that the physiological significance of IL-1 as a tissue CRF is indeed questionable and should be further clarified. PMID- 3035405 TI - Reduced S-adenosylmethionine:protein-lysine N-methyltransferase activity (protein methylase III) in shiverer mutant mouse brain. AB - Mice with the dysmyelinating mutation shiverer were studied by measuring the activity of two protein methylases and myelin marker enzymes in the brain. It was observed that S-adenosylmethionine:protein-lysine N-methyltransferase (protein methylase III, EC. 2.1.1.43) activity is significantly reduced in phenotypically affected homozygous shiverer (shi/shi) mutant mouse brain compared to the unaffected heterozygous littermate brain. This reduction in enzyme activity is manifested mainly by reduced formation of trimethyllysine during the in vitro methylation of histone. In contrast, myelin marker enzymes such as 2',3'-cyclic nucleotide 3'-phosphohydrolase and 5'-nucleotidase as well as S-adenosyl methionine:protein-carboxyl O-methyltransferase (protein methylase II, EC. 2.1.1.24) activities were not significantly affected in these strains of mice. PMID- 3035408 TI - Dopaminergic neurones in the zona incerta exert a stimulatory control on gonadotrophin release via D1 dopamine receptors. AB - The zona incerta (ZI) is a site of dopamine nerve terminals and part of the incertohypothalamic tract (I-H). Previous findings indicate that dopamine in the ZI has a stimulatory control on the release of luteinizing hormone (LH) and occurrence of ovulation. The effect of acute administration into anaesthetised rats of selective D1 and D2 dopamine agonists and antagonists injected into the ZI on plasma luteinizing hormone (LH) and on the occurrence of ovulation has now been investigated. It was found that bilateral injections on the day of pro oestrus of a selective D1 antagonist, Sch 23390, inhibited ovulation at 10 micrograms/side/rat. Unilateral injections of a selective D1 agonist, SKF 38393, at 10 micrograms/rat stimulated a significant rise in plasma LH concentration in ovariectomised oestrogen-primed rats, and this was partially reversed by systemic pre-treatment with Sch 23390. The selective D2 agonist, LY 171555, and D2 antagonists, sulpiride and domperidone, had no effect on plasma LH levels or ovulation. This indicates that D1 receptors (but not D2 receptors) in the ZI are involved in the control of gonadotrophin release and may have a physiological function in reproductive processes. PMID- 3035409 TI - Histaminergic neuromodulation of the release of vasopressin. AB - In an attempt to clarify the nature of histaminergic neuromodulation of the vasopressinergic system, several studies under different experimental paradigms were carried out. L-Histidine loads (8 mmol/kg, i.p.) induced a marked increase in histamine (HA) in the anterior (AHR) and posterior (PHR) hypothalamic regions, the median eminence (ME) and adenohypophysis (Ah) with no apparent effect on the concentration of HA in the neurohypophysis (Nh), as measured by high-performance liquid chromatography. These findings correlated with decreases in vasopressin (VP) levels in the AHR and ME, accompanied by increases of the neuropeptide in the PHR and Ah. Intraperitoneal injections of HA (6 mumol/kg), resulted in a significant (p less than 0.005) rise in VP levels in the PHR, ME and Ah. HA induced an elevation of VP in the prefrontal cortex (PFC) from 6.23 +/- 2.02 to 43 +/- 4.05 microU/mg, as well as a 60% reduction in neurohypophyseal VP. These HA-induced VP responses were abolished by both mepyramine (3 mumol/kg) and famotidine (4 mumol/kg) in the PHR and PFC. Mepyramine suppressed the HA-induced VP response in the Ah and enhanced it in the Nh, while famotidine did the opposite. When alpha-fluoromethylhistidine (FMH), an irreversible inhibitor of histidine decarboxylase, was administered at doses of 100 mg/kg/day (i.p.), hypothalamic HA levels fell by 40-45% after 1 h, by 50% after 3 h, and by 65-80% after 24 h in adrenalectomized rats. In the same conditions, but after a week of treatment with FMH, the VP response to adrenalectomy was clearly impaired.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3035410 TI - Effects of castration or testosterone implants upon pituitary function in hypogonadal mice bearing normal foetal preoptic area grafts. AB - Grafts of normal mouse preoptic area (POA) tissue into the third ventricle of gonadotrophin-releasing hormone (GnRH)-deficient hypogonadal (hpg) mice resulted in an elevation of pituitary GnRH receptors, an increased synthesis of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) by the pituitary gland, an elevation of gonadal LH receptors and in the stimulation of steroidogenesis and spermatogenesis in the testis. In normal mice both castration or the subcutaneous implantation of testosterone capsules for 10 days reduced GnRH receptors, pituitary LH and FSH content, and the latter treatment also caused a 50% reduction in testicular LH receptors. In hpg mice bearing POA grafts testosterone implants failed to affect any of the above parameters, and castration failed to affect pituitary gonadotrophin hormone content, although there was a slight reduction in pituitary GnRH receptors after castration. These experiments suggest that neither the pituitary gonadotroph, nor the GnRH neurone represent major sites for the direct negative feedback of testosterone upon gonadotrophic hormone secretion in male mice. PMID- 3035411 TI - Stress-induced changes in the function of the parasympathetic nervous system are mimicked by blocking GABA in the CNS of the cat. AB - The purpose of this study was to determine the effect of blockade of receptors for gamma-aminobutyric acid (GABA) in the forebrain, on vagal activity to the stomach and heart. This was done by injecting bicuculline (50 micrograms) into the lateral ventricle of the brain and restricting the drug to the forebrain ventricles by cannulating the cerebral aqueduct. Studies were performed in chloralose-anesthetized cats and gastric motility was monitored using extraluminal force transducers, sutured to the antrum and pylorus. Cardiac vagal activity was determined by noting the sinus bradycardia that developed from activation of the baroreceptor reflex induced by phenylephrine. Administration of bicuculline into the lateral ventricle of 7 animals produced increases in the minute motility index of 5.3 +/- 0.8 (antrum) and 13.9 +/- 2.1 (pylorus). This was associated with inhibition of baroreceptor-induced vagal bradycardia (i.e. 38 +/- 6.4 beats/min before bicuculline and -7.7 +/- 5.7 beats/min after bicuculline). These data indicate that a GABAergic mechanism in the forebrain may be important for controlling vagal outflow to both the stomach and the heart. PMID- 3035413 TI - Cannabimimetic activity of cannabinol in rats and pigeons. AB - The present experiments examined the cannabimimetic potential of cannabinol on its own as well as its interaction with delta 9-tetrahydrocannabinol (delta 9 THC) in vivo. Thus, cannabinol, as evaluated by repeated test procedures using drug-discrimination methodology (Hiltunen and Jarbe, 1986; Jarbe, Swedberg and Mechoulam, 1981) was found to substitute for the stimulus effects induced by delta 9-THC in both rats (delta 9-THC cue = 3 mg/kg) and pigeons (delta 9-THC cue = 0.56 mg/kg), though, relatively larger doses were required in the latter species (ED50 for cannabinol being 8.4 mg/kg, and 14.1 mg/kg at the time of maximum effect in rats and pigeons, respectively). When administered together, cannabinol added to the cue effects of delta 9-THC as shown by an increase in the percentage of responding appropriate to the drug, the effect appearing more pronounced in rats than in pigeons; no apparent change in the duration of the effect was indicated by the results obtained. Unconditioned effects of drug, as defined by rectal temperature recordings and open-field activity (ambulation, rearing, latency, circling, grooming, defecation and urination), as well as assessment of vocalization in rats were in keeping with the suggestion that the pharmacological profile of cannabinol is similar, though not necessarily identical, to that of tetrahydrocannabinols, such as delta 9-THC. PMID- 3035412 TI - Effects of lithium in vitro and ex vivo on components of the adenylate cyclase system in membranes from the cerebral cortex of the rat. AB - The effects of lithium on the activity of adenylate cyclase stimulated by hormones, which act via the stimulatory guanine nucleotide binding subunit (Ns), by forskolin, which acts at the catalytic subunit, and by guanyl-5'-yl imidodiphosphate (GppNHp), which locks the enzyme into a permanently active state, have been compared in a preparation of membranes from the cerebral cortex of the rat. Lithium ions (Li+) in vitro at 2-4 mM inhibited cyclase stimulated by isoproterenol and forskolin, but had no effect on the inhibition induced by met enkephalin of the enzyme stimulated by forskolin, mediated by the inhibitory guanine nucleotide binding subunit (Ni). Inhibition of the activity stimulated by forskolin and GppNHp was competitive with magnesium (Mg++). In a preparation of slices of cerebral cortex Li+ at 1-2 mM inhibited accumulation of cyclic AMP stimulated by forskolin in a non-competitive manner. In a preparation of membranes from the caudate nucleus, Li+ at 2-4 mM inhibited dopamine-stimulated adenylate cyclase, but this effect was not observed in the presence of additional sodium (Na+). Membranes prepared from animals fed with Li+ to give a mean serum level of 0.52 mM and a mean brain level of 1.32 mM, showed a reduced response to manganese (Mn++), forskolin, isoproterenol and GppNHp in the cerebral cortex, but no change in the degree of activation of the enzyme by either dopamine or forskolin, or the degree of inhibition by met-enkephalin, in the caudate nucleus.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3035414 TI - Enhancement of the hypothermic response of mice to delta-9-tetrahydrocannabinol by subhypothermic doses of chlorpromazine and phentolamine. AB - Pretreatment with subhypothermic doses of chlorpromazine, given directly into the IIIrd cerebral ventricle via a chronically implanted cannula (50 micrograms) or subcutaneously (0.75 mg/kg), was found to enhance the hypothermic response to delta-9-tetrahydrocannabinol (THC: 5 20 mg/kg i.p.) in unrestrained adult male MF1 mice, kept at 22 degrees C. Subcutaneous pretreatment with a subhypothermic dose of phentolamine (30 mg/kg) had a similar effect, whereas pretreatment with desipramine (10 mg/kg s.c.), mepyramine (2.3 and 11.5 mg/kg s.c.), methysergide (2 mg/kg s.c.), pimozide (1 and 5 mg/kg s.c.) or lignocaine (50 mg/kg s.c.), had no effect. Intracerebroventricular pretreatment with phentolamine was also without effect and it is concluded that this drug interacts with THC at some site located outside the brain. Since, in mg/kg terms, chlorpromazine was more potent in enhancing THC-induced hypothermia when given subcutaneously than when injected into the IIIrd ventricle, it too may interact with THC at a peripheral site. Indeed, chlorpromazine and phentolamine may both increase the hypothermic response to THC by antagonizing alpha-adrenoceptors on cutaneous blood vessels, thereby decreasing the capacity of animals to minimise peripheral blood flow by vasoconstriction. Alternatively, since the distribution of chlorpromazine within the brain may well have been less efficient after intraventricular than after subcutaneous injection, the possibility remains that chlorpromazine interacted centrally with THC. PMID- 3035415 TI - Enhancement of the in vivo binding of [3H]flunitrazepam by the atypical neuroleptic, clozapine. AB - Modulation of the benzodiazepine receptor/GABA receptor/chloride ionophore complex in vivo involves a number of intricate regulatory interactions between the three components of the receptor complex. One way to assess these potential interactions involves the in vivo labelling of the benzodiazepine receptor with [3H]flunitrazepam. In these studies, we used this approach to demonstrate that the neuroleptic, clozapine, increases [3H]flunitrazepam binding in mouse brain in a bicuculline-reversible manner. This potentiation of benzodiazepine binding was not antagonized by picrotoxin and was found to result from a slower dissociation of [3H]flunitrazepam from the benzodiazepine receptor. These data suggest that clozapine acts to increase [3H]flunitrazepam binding via a GABAergic mechanism, independent of the chloride channel. PMID- 3035416 TI - Selective inhibition of nociceptive flexion reflex discharge by the kappa agonist bremazocine. AB - In unanaesthetized decerebrate spinal cats, bremazocine (0.012-0.2 mg/kg, i.v.) selectively inhibited the late C-fibre reflex discharge, recorded in sectioned lumbo-sacral ventral root filaments, after supramaximal electrical stimulation of the ipsilateral sural or common peroneal nerve. This action was naloxone reversible. The results suggest that activation of kappa opioid receptors in the spinal cord inhibits the integration of nociceptive, but not locomotor, flexion reflexes. PMID- 3035418 TI - Malignant fibrous histiocytoma of the clivus: case report. AB - A case of malignant fibrous histiocytoma of the clivus in a 2 1/2-year-old boy is reported. There are no prior reports of this tumor in this location. The child was treated with operation, radiation therapy, and chemotherapy. The tumor recurred locally 21 months later. The recurrence was palliated by operation and chemotherapy, permitting an additional 20-month survival. PMID- 3035417 TI - Antagonism of ethanol-induced depressant effects by 4-aminopyridine in the central nervous system of the rat. AB - Intraperitoneal injection of ethanol (2 g/kg) produced significant motor impairment in rats, as measured by performance on the tilting plane. Administration of 3 mg/kg 4-aminopyridine (4-AP) antagonized the depressant effect of ethanol on motor performance. Using slices of hippocampus, in vitro, 4 aminopyridine (10-100 microM) also antagonized the ethanol-induced depressant effect on orthodromically-elicited population spikes in the CA1 pyramidal cell layer. This antagonism appears to result from the ability of 4-aminopyridine to enhance release of transmitter in both excitatory and inhibitory neurones. Due to a number of unwanted side effects, further evaluation of 4-aminopyridine and its analogues needs to be done before it can be considered useful in the management of acute intoxication with ethanol. PMID- 3035419 TI - Comparing oral lithium carbonate and intraperitoneal lithium chloride chronic administrations on rats' activity levels. AB - This study compares the effects produced by two modalities of lithium administration on rats' activity levels. Lithium was administered for 21 days either as carbonate in the diet (2 g in 2 liters of water and 1,500 g lab chow) or as chloride through single daily intraperitoneal injections (1 mEq/kg LiCl). Both treatments resulted in similar decreases in spontaneous activity and exploratory behavior, as recorded in an open field with a hole-board. Neither treatment influenced the amount of food consumed on a daily basis nor the rate of growth as manifested by weight gains over the period considered, suggesting that these treatments were not adversely affecting the animals' health. Both treatments increased water intake over control levels, this effect being most marked with the diet lithium carbonate administration, particularly during the second week of treatment. This latter modality of lithium administration, but not the injections of lithium chloride, significantly increased NaCl intake over control levels. PMID- 3035420 TI - Multiple sites for the initiation of action potentials in neurons of the inferior mesenteric ganglion of the guinea-pig. AB - Responses to nerve stimulation were recorded with intracellular microelectrodes from neurons in the inferior mesenteric ganglion of the guinea-pig. An event was recorded which was subthreshold for a somatic action potential but which had a short rise time and a rapid initial repolarization that gave the event a small but well-defined peak. This event was termed a "partial spike." During repeated stimulation of the same nerve trunk, the fluctuations in the amplitude of the partial spike were small compared to those of evoked synaptic potentials. Stimulation of different nerve trunks evoked partial spikes of different amplitudes. When different nerve trunks were stimulated at short intervals between stimuli, one partial spike could occlude another partial spike. Antidromic responses could not be blocked by a preceding partial spike. This suggests that partial spikes are not initial segment spikes. Tubocurarine reversibly abolished partial spikes which is indicative of a synaptic origin. It is concluded that partial spikes result from action potentials initiated by synaptic potentials in the dendrites and which fail to generate somatic action potentials. PMID- 3035421 TI - GABAA and GABAB receptor site distribution in the rat central nervous system. AB - An autoradiographic procedure has been used to determine the quantitative distributions of gamma-aminobutyric acid (GABAA and GABAB) receptor subtypes in rat brain. Although the concentrations of both receptor binding sites were similar in some brain regions GABAA sites generally outnumbered GABAB sites. The highest concentration of GABAA sites were detected in the frontal cortex, the granule cell layer of the cerebellum, the olfactory bulb and the thalamic medial geniculate. The highest concentration of GABAB sites occurred in the molecular layer of the cerebellum, the interpeduncular nucleus, frontal cortex, anterior olfactory nucleus and thalamic nuclei. In addition the globus pallidus, temporal cortex, lateral posterior thalamus, superior colliculus, pontine nucleus, raphe magnus, spinal trigeminal tract and substantia gelatinosa contained significantly more GABAB sites than GABAA sites. The physiological and pharmacological significance of this heterogeneity has yet to be determined. PMID- 3035422 TI - A quantitative light microscopic analysis and ultrastructural description of cholecystokinin-like immunoreactivity in the spinal trigeminal nucleus of the rat. AB - The spinal trigeminal nucleus is involved in orofacial sensory transmission. Cholecystokinin octapeptide has been identified in axons in this nucleus and appears to play a role in the transmission of orofacial sensation from the trigeminal ganglia to the spinal trigeminal nucleus. Although cholecystokinin has been reported in axonal processes within the spinal trigeminal nucleus at the light microscopic level, nothing is known about the synaptic relationships of these cholecystokinin axons. The goals of this study were to quantitatively determine the volume fraction of cholecystokinin-like immunoreactive cell bodies and fibers in the three subnuclei of the spinal trigeminal nucleus, to provide the first ultrastructural description of cholecystokinin-like immunoreactive processes within these subnuclei and to analyse the synaptic relationships of cholecystokinin-like immunoreactive processes within the spinal trigeminal nucleus neuropil. Cholecystokinin-like immunoreactivity was localized by the peroxidase-antiperoxidase method or the peroxidase labeled, avidin-biotin technique and quantified at the light microscopic level by point counting. Immunoreactive fibers were present in all three subnuclei, but the greatest volume fraction of immunoreactive axons was obtained in laminae I and II of the nucleus caudalis. No immunoreactive cell bodies were evident in any of the subnuclei. The majority of immunoreactive profiles in all three subnuclei were identified ultrastructurally as axon terminals that contained both small and medium sized agranular vesicles and infrequently, large dense core vesicles. These immunoreactive terminals were usually found in close contact with non immunoreactive dendrites with which they were observed to form asymmetric synapses. Immunoreactive terminals were occasionally observed to contact the cell bodies of large non-immunoreactive neurons on the border of laminae I and II in the nucleus caudalis. These results indicate that cholecystokinin-like immunoreactive processes are present throughout the spinal trigeminal nucleus, and in nucleus caudalis show a distribution similar to that reported for the spinal cord dorsal horn. Immunoreactive axons make synaptic contact with both the dendrites and perikarya of spinal trigeminal nucleus neurons. No axoaxonic synapses were observed. These findings suggest that cholecystokinin plays an important role in spinal trigeminal nucleus function. The possible colocalization of cholecystokinin and substance P in the spinal trigeminal nucleus, and the possible role of cholecystokinin in attenuating the action of opioids in the spinal trigeminal nucleus are also discussed. PMID- 3035423 TI - A subpopulation of preganglionic parasympathetic neurons in the rat contain adenosine deaminase. AB - Immunohistochemical staining and retrograde fluorescent tracing techniques were used to demonstrate the presence of adenosine deaminase in preganglionic parasympathetic neurons. Both brainstem and sacral spinal cord parasympathetic nuclei were found to contain a subpopulation of neurons immunoreactive for adenosine deaminase. Immunostaining of preganglionic neurons in brainstem was restricted to a group of cells which were shown by retrograde tracing with Fast Blue to project exclusively to the sphenopalatine ganglion. This group was defined as the lacrimo-nasopalatine parasympathetic nucleus. Neurons in all other cranial preganglionic centers were devoid of adenosine deaminase immunoreactivity. In spinal cord adenosine deaminase-immunoreactive neurons were found in the intermediolateral gray matter in the region of the sacral parasympathetic nucleus. Injections of Fast Blue into the pelvic ganglion labeled large numbers of neurons in this nucleus, only some of which contained adenosine deaminase. The majority of neurons immunoreactive for adenosine deaminase were also shown to be immunoreactive for choline acetyltransferase in both brainstem and sacral parasympathetic nuclei. The present results show that a subclass of preganglionic parasympathetic neurons are among the few structures in the central nervous system that express what appear to be high levels of adenosine deaminase. This observation together with evidence suggesting that purines serve as neurotransmitters in some sacral parasympathetic neurons supports the notion that adenosine deaminase may constitute a marker for adenine nucleoside and/or nucleotide neurotransmission. PMID- 3035426 TI - Familial progressive neuronal disease and chronic idiopathic intestinal pseudo obstruction. AB - Chronic idiopathic intestinal pseudo-obstruction (CIIP) is characterized by recurrent episodes of bowel obstruction without mechanical cause. In five members of two Jewish-Iranian families, CIIP was associated with progressive neuronal disease, starting before age 30, with ophthalmoplegia, sensorimotor peripheral neuropathy, and hearing loss. There was no evidence of CNS involvement. The pattern suggested autosomal recessive inheritance. PMID- 3035425 TI - Angiotensin converting enzyme in rat brain visualized by quantitative in vitro autoradiography. AB - Angiotensin converting enzyme was localized in rat brain by quantitative in vitro autoradiography using an [125I]labelled converting enzyme inhibitor called "351A". This radioligand was found to bind with high affinity and specificity to angiotensin converting enzyme. Very high levels of converting enzyme were observed in the ventricular choroid plexus, ependyma of all ventricles and large and medium blood vessels, subfornical organ, and organum vasculosum of the lamina terminalis. High levels of converting enzyme were found in the basal ganglia including caudate putamen, nucleus accumbens, globus pallidus, entopenduncular nucleus and substantia nigra pars reticulata. The neurosecretory nuclei, paraventricular nucleus and supraoptic nucleus, as well as the median eminence and posterior pituitary displayed high levels of the enzyme. In the amygdala, basolateral, lateral, basomedial, medial and anterior cortical nuclei showed moderate converting enzyme activity. The medial habenula and molecular layer of the dentate gyrus showed high levels of activity. In the cerebellum, dense labelling was observed in the Purkinje cell layer. Moderate levels of converting enzyme occurred in the gelatinosus subnucleus of the caudal part of the nucleus of the spinal tract of the trigeminal. There was a close correspondence between the distribution of angiotensin converting enzyme and angiotensin II in the neurosecretory nuclei (paraventricular and supraoptic nuclei) and median eminence and this suggests a role of angiotensin converting enzyme in the production of angiotensin II in this system. There was also a good correspondence between the distribution of angiotensin converting enzyme and angiotensin II in the subfornical organ, median preoptic nucleus, and organum vasculosum of the lamina terminalis, structures abutting the anterior wall of the third ventricle which are implicated in fluid and electrolyte homeostasis. A striking discrepancy occurs in the basal ganglia which is reported to contain very little angiotensin II or angiotensin II receptors but is very rich in angiotensin converting enzyme. It is concluded that the enzyme may act to convert circulating angiotensin I to angiotensin II in circumventricular organs; generate intraneuronal angiotensin II in pathways such as the hypothalamic-hypophyseal tract; and process neuropeptides other than angiotensin II in regions such as basal ganglia. PMID- 3035424 TI - Cholinergic innervation of ferret visual system. AB - The distribution of acetylcholinesterase and choline acetyltransferase in primary visual areas of adult pigmented ferret was determined with cholinesterase histochemistry and choline acetyltransferase immunohistochemistry. In all visual areas the distribution of acetylcholinesterase in the neuropil closely matches that of choline acetyltransferase. In the cerebral cortex acetylcholinesterase and choline acetyltransferase are associated with axons found in every cortical layer and in the white matter. Area 17, identified by Nissl architectonics and cytochrome oxidase histochemistry, is distinguished by having a relatively low density of choline acetyltransferase- and acetylcholinesterase-stained axons in layer IV. Certain cortical non-pyramidal cell types show moderate staining for acetylcholinesterase after relatively long incubations, but no choline acetyltransferase-positive cells are observed in the cortex. In the lateral geniculate nucleus and superior colliculus the levels of choline acetyltransferase and acetylcholinesterase are considerably higher than in cerebral cortex, and choline acetyltransferase-stained axons there display prominent varicosities. The distribution of choline acetyltransferase and acetylcholinesterase in the neuropil of lateral geniculate nucleus and superior colliculus of ferret shows marked laminar variation. For instance, in the lateral geniculate nucleus, the levels of acetylcholinesterase and choline acetyltransferase in the "On" sublaminae of laminae A and A1 are higher than the "Off" sublaminae. In the superficial layers of the superior colliculus the levels of choline acetyltransferase and acetylcholinesterase are highest in the stratum zonale and lowest in the stratum opticum; in the intermediate gray layer of the superior colliculus acetylcholinesterase- and choline acetyltransferase-stained fibres are distributed into dense patches. As in cortex, choline acetyltransferase-positive cell bodies are not found in the lateral geniculate nucleus or superior colliculus, and acetylcholinesterase-stained cell bodies are visible only after long incubations. Cell bodies staining positively for choline acetyltransferase are found in a satellite of the superior colliculus, the parabigeminal nucleus. PMID- 3035427 TI - Huntington's disease: increased number and altered regulation of benzodiazepine receptor complexes in frontal cerebral cortex. AB - The properties of benzodiazepine receptors in samples of six areas of cerebral cortex from patients and controls matched with respect to age, sex, and postmortem delay have been studied by in vitro radioligand binding techniques. A statistically significant 30 to 40% increase in the number of benzodiazepine and allosterically linked GABA-A receptors is observed in the midfrontal cortex (A9/A10), but in no other cortical region examined. The degree of enhancement of [3H]diazepam binding by a given dose of GABA or pentobarbital is significantly reduced in Huntington's disease midfrontal cortex. As in the case of receptor number changes, other cortical regions examined show less prominent changes in regulation of benzodiazepine binding by GABA or pentobarbital. Direct identification of the benzodiazepine binding subunit by photoaffinity labeling with [3H]flunitrazepam reveals a single species of apparent molecular weight 51 kD in patients and controls, suggesting that gross changes in structure of the benzodiazepine binding subunit do not account for the observed alterations in benzodiazepine receptor regulation. PMID- 3035428 TI - Task-dependent gating of somatosensory transmission in two different motor tasks in man: falling and writing. AB - The cerebral potentials induced by an electrical stimulus (median nerve or finger) were recorded over the central region of the scalp and were analysed during falling onto the extended arms or during writing to investigate the influence of different motor tasks on the transmission of a synchronous afferent volley to the brain. During both falling (before landing) and writing, the first peaks (20-40 ms) were reduced. Later peaks (60-200 ms) were enhanced during writing but reduced during falling. A reduction of the first peak was also obtained after ischaemic blockade of group I afferents, suggesting that the cerebral transmission of group I afferents is inhibited during falling and writing. The subjects reported a corresponding reduction in the perception of the stimulus during falling. During writing, however, the large late waves indicate a task specific processing of the remaining afferent volley. Such a gating of sensory information to the brain is assumed to play a functional role in the respective motor tasks. PMID- 3035429 TI - Plasma immunoreactive atrial natriuretic factor is inhibited by selective blockade of alpha 2-adrenergic receptors in conscious Sprague-Dawley rats. AB - Effects of alpha 2-adrenergic receptors and calcium channel blockade on basal and clonidine-stimulated immunoreactive atrial natriuretic factor (IR-ANF) in conscious Sprague-Dawley rats were evaluated. Clonidine was injected intravenously (i.v.) in a dose of 50 micrograms. Yohimbine and verapamil were used as a pretreatment, with clonidine in a dose of 50 micrograms and 0.5 mg respectively. The effects of yohimbine (1, 20, 50 micrograms) and verapamil (0.5 mg) on basal IR-ANF were also studied. Plasma IR-ANF was measured by radioimmunoassay with prior extraction on heat-activated Vycor glass. Clonidine injection in a dose of 50 micrograms caused a marked increase of plasma IR-ANF from 34.0 +/- 7.0 pg/ml (mean +/- S.E.M.) to 457.1 +/- 66.3 pg/ml. Clonidine stimulated ANF secretion was partially inhibited by yohimbine from 457.1 +/- 66.3 pg/ml (mean +/- S.E.M.) to 99.9 +/- 23.1 pg/ml. Moreover, yohimbine in highest doses (50 micrograms) decreased the basal plasma IR-ANF from 34.0 +/- 7.0 pg/ml (means +/- S.E.M.) to 6.8 +/- 3.6 pg/ml. Verapamil did not alter basal and clonidine stimulated IR-ANF. These results indicate the important role played by alpha 2-adrenergic receptors in mediating ANF release. PMID- 3035430 TI - Age-dependent and selective binding of beta-bungarotoxin to GABAergic neurons in the rat cerebellum. AB - beta-Bungarotoxin (BuTx)-binding cells were immunocytochemically examined in the developing rat cerebellum. The tissue was incubated with BuTx and then immunostained with antiserum against its toxoid. On postnatal day 6, only Golgi cells were positive for immunoreaction. Immunoreactive Golgi cells were reduced in number on day 15 and disappeared on day 25. On day 15, Purkinje cells were strongly stained, while some basket and stellate cells stained weakly. On day 25 and in adult, basket and stellate cells were more immunoreactive than Purkinje cells. Thus, age-dependent and selective binding of BuTx was restricted to gamma aminobutyric acid (GABA)ergic neurons. PMID- 3035432 TI - Spinal cord alpha 2-adrenoceptors may be located postsynaptically with respect to primary sensory neurons: destruction of primary C-afferents with neonatal capsaicin does not affect the number of [3H]clonidine binding sites in mice. AB - Mice were treated with 50 mg/kg capsaicin on the second day of life, which causes a permanent and selective loss of a large proportion of afferent C-fibers. Two months later the spinal cords were removed and the content of alpha 2 adrenoceptors assayed by radioligand binding with [3H]clonidine. In these tests the content of alpha 2-adrenoceptors was found to be 568 +/- 27 fmol/mg protein in the capsaicin-treated animals and it was virtually the same -596 +/- 9 fmol/mg protein - in control animals. The affinity of [3H]clonidine for the alpha 2 adrenoceptor was not affected by capsaicin (Kd = 3.25 +/- 0.29 nM for capsaicin and 3.23 +/- 0.32 nM for the controls). In functional tests clonidine applied s.c. induced a marked reduction in the pain sensitivity to capsaicin. The results indicate that spinal cord alpha 2-adrenoceptors are located postsynaptically with respect to the primary sensory neurons and potentially that stimulation of these receptors by alpha 2-adrenoceptor agonist mediates inhibition of nociceptive transmission. PMID- 3035431 TI - Effects of bicarbonate on glial cell membrane potential in Necturus optic nerve. AB - Intracellular electrodes were used to continuously monitor the membrane potential of glial cells in the isolated Necturus optic nerve. Addition of up to 10 mM extracellular bicarbonate (with CO2), at constant pH, produced a hyperpolarization of up to 10 mV (with a time course almost as fast as that of a K+ depolarization) that returned toward baseline during the following 2-15 min. Upon bicarbonate withdrawal, the potential transiently became more positive. The bicarbonate effects were magnified when the K+ conductance was decreased and the cell depolarized by the addition of barium. Similar bicarbonate effects were observed in Cl- free solutions. These results suggest to us that: glial cells have a bicarbonate permeability of the same order as that to K+ and glial cells buffer transient changes in acid base balance in the neuronal microenvironment at the expense of their internal pH. PMID- 3035433 TI - The distribution of GABA and glycine response in the mouse brain using Xenopus oocytes. AB - Injection of mRNA from different regions of the central nervous system (CNS) of the mouse into the Xenopus oocyte caused a difference in the relative glycine to gamma-aminobutyric acid (GABA) response of the oocyte. Glycine caused the response in oocytes injected with brainstem mRNA but hardly in oocytes injected with cerebrum mRNA and in oocytes injected with cerebellum mRNA. In contrast, GABA caused the response in oocytes injected with mRNA from each of the 3 parts of the CNS. These obvious regional differences may reflect the distribution of the receptors to each plausible neurotransmitter in the CNS. PMID- 3035434 TI - The chloride channel blocking agent, t-butyl bicyclophosphorothionate, binds to the gamma-aminobutyric acid-benzodiazepine, but not to the glycine receptor in rodents. AB - The inhibitory neurotransmitters glycine and gamma-aminobutyric acid (GABA) both activate transmembrane chloride channels of similar physical characteristics. A common ion channel component has therefore been postulated for both the glycine and GABA receptor proteins. Different convulsant drugs as picrotoxin and t-butyl bicyclophosphorothionate (TBPS) have been reported as channel-blocking ligands of the GABA receptor. Here, we show that the distribution of [35S]TBPS binding sites parallels the binding of the GABA receptor ligand [3H]flunitrazepam, but not that of the glycine receptor antagonist [3H]strychnine. Binding was examined in membrane fractions from different regions of the rat CNS and of the mutant mouse spastic, an animal deficient in glycine receptors. Also, affinity purification of the glycine receptor on aminostrychnine-agarose resulted in almost complete removal of [35S]TPBS binding sites from the receptor preparation. It is concluded that TBPS selectively binds to the GABA, but not glycine, receptor chloride channel complex. PMID- 3035435 TI - Loss of GABAergic neurons in medial septum after fimbria-fornix transection. AB - Neurons in the medial septum of the rat brain undergo retrograde degeneration after transection of their projection to the hippocampal formation, the fimbria fornix. This cell death has been characterized for both Nissl-stained neurons and acetylcholinesterase-stained neurons. The major cell type in the medial septum is GABAergic, and many of these GABAergic neurons project to the hippocampal formation. Because the fimbria-fornix transection causes more neuronal death than can be accounted for by the loss of cholinergic neurons, we have sought to determine if the GABAergic neurons undergo a cell death similar to that reported for the cholinergic neurons. We report here that GABAergic neurons are indeed lost after the transection but the time course is considerably slower than that for the cholinergic neurons. PMID- 3035437 TI - Inhibitory effects of neuropeptide Y on cyclic AMP accumulation in slices of the nucleus tractus solitarius region of the rat. AB - The effects of neuropeptide Y (NPY) on cyclic AMP (cAMP) accumulation in slices of the dorsal midline area of the caudal part of the medulla oblongata containing the nucleus tractus solitarius (nTS) have been studied. Neuropeptide Y (30 and 300 nM) significantly reduced the [3H]cAMP accumulation induced by forskolin and phorboldibutyrate. Similar results were obtained after incubation with the alpha adrenoceptor agonist clonidine (1 microM). These results indicate that stimulation of the NPY receptors and alpha-adrenoceptors in the nTS region may cause inhibition of the adenylate cyclase. Such a mechanism may at least partly underlie the centrally mediated hypotensive effects of the costored transmitters adrenaline and NPY. PMID- 3035436 TI - Distribution of neurotensin receptors in the primate hippocampal region: a quantitative autoradiographic study in the monkey and the postmortem human brain. AB - The distribution of [3H]neurotensin ([3H]NT) binding sites in the monkey and the postmortem human brain was studied by using quantitative in vitro receptor autoradiography. Biochemical experiments carried out on tissue sections of the monkey hippocampus showed that the binding of [3H]NT was saturable, reversible and of high specificity. The hippocampal [3H]NT binding was displaced by fragment NT 8-13 but not fragment NT 1-8 of the peptide. The anatomical analysis showed a highly heterogeneous distribution of [3H]NT binding sites within both the monkey and the human hippocampal region. In both species the highest density of [3H]NT binding sites was found in the presubiculum (rank order of binding density: layer 2 greater than 6 greater than 1 greater than 3, 4, 5 in both monkey and man) and the entorhinal area (monkey: layer 4 greater than 6 greater than 5 greater than 1 greater than 2 greater than 3; human: layer 1 = 2 greater than 5 greater than 3). The subiculum and Ammon's horn were relatively poor in [3H]NT binding sites in both species. In the area dentata the highest density of [3H]NT binding sites was found in the hilar region. PMID- 3035438 TI - Effects of forskolin on spontaneous behavior, rectal temperature and brain cAMP levels of rats: interaction with rolipram. AB - The effects of systemically administered forskolin, a direct activator of the catalytic subunit of adenylate cyclase, on locomotor activity, rectal temperature and the incidence of grooming and head twitches were studied in rats. Forskolin, 0.1-25 mg/kg, decreased locomotor activity and lowered rectal temperature. The incidence of grooming and head twitches increased dose-dependently after forskolin, 6.25 25 mg/kg. The combination of a threshold dose of forskolin with a threshold dose of the cAMP-selective phosphodiesterase (PDE) inhibitor rolipram resulted in a marked, statistically significant enhancement of the behavioral, hypothermic and brain cAMP elevating effect. The present findings support the assumption that enhanced brain cAMP availability is associated with a characteristic behavioral syndrome in rats as was previously shown with dibutyryl cAMP or neurotropic cAMP-selective PDE inhibitors like rolipram. Ro 20-1724 or ICI 63 147. PMID- 3035440 TI - GABAergic nerve terminals in rat hippocampus possess alpha 2-adrenoceptors regulating GABA release. AB - Noradrenaline enhanced in a concentration-dependent way the basal release of endogenous GABA from superfused rat hippocampus synaptosomes. The alpha 2 adrenoceptor antagonist yohimbine prevented the releasing effect of noradrenaline while the alpha 1-adrenoceptor antagonist prazosin was ineffective. It is concluded that GABAergic nerve terminals in rat hippocampus possess adrenoceptors of the alpha 2-subtype whose activation causes enhancement of GABA release. PMID- 3035439 TI - Transformation of glial cells in mouse embryonic brain cells in vitro with simian virus 40. AB - Embryonic (E10) mouse cerebellum and spinal cord cells were cultured and infected with simian virus 40 (SV40). In all infected dishes, rapid proliferations (a dividing time less than 18 h) of non-neuronal cells were detected. Three populations of those cells, SVa, SVb and SVc, could be passaged more than 10 times. They had flat morphologies like young astrocytes and, indeed, showed immunoreactivities to glial fibrillary acidic protein (GFAP), a marker for the astrocyte. However, they had an ability to form colonies in a soft agarose medium. The large-T antigen could be detected in nuclei of the three cell populations by a fluorescent antibody test and immunoprecipitation. No transformation of neuronal cells was detected, nevertheless there were neuronal cells still possessing dividing capacities in the culture preparation. PMID- 3035442 TI - Herpes simplex pericarditis in AIDS. PMID- 3035441 TI - Demography of HIV infections among civilian applicants for military service in four counties in New York City. PMID- 3035443 TI - Exercise and small intestinal transit. AB - The transit of 99Tcm-labelled particles through the small intestine has been monitored in nine healthy subjects undergoing three levels of exercise. The mean +/- 1S.D. small intestine transit times under conditions of minimal, moderate and strenuous activity were 4.5 +/- 1.7 h, 5.4 +/- 2.5 h and 4.1 +/- 1.8 h, respectively. These findings indicate that the bioavailability of drugs from controlled release preparations passing through the small intestine is unlikely to be affected by variations in normal daily activity. PMID- 3035444 TI - Preparation of 99Tcm (V) DMSA. PMID- 3035445 TI - [Development, responsibility and significance of after-care treatment]. PMID- 3035446 TI - [Hyaluronidase as an adjuvant to cystostatic chemotherapy in glioblastoma]. PMID- 3035448 TI - Granular cell tumor (myoblastoma) of the eyebrow. AB - Granular cell tumor (Myoblastoma) is a tumor of controversial nature that rarely appears in the ocular adnexa. A 37-year-old man with such a tumor in his right eyebrow is reported, and the relevant literature is reviewed. PMID- 3035447 TI - [Chemotherapy of non-small cell bronchial cancer with mitomycin C, vindesine and cisplatin. Results of a phase II study]. AB - Forty-five patients (39 male, 6 female) with inoperable non-small-cell lung cancer were treated with combined mitomycin C, vindesine and cisplatin. All patients had measurable disease and had no previously received chemotherapy. The performance status of all patients was over 60%. Twenty-one patients had limited, 24 extensive disease. The overall remission rate was 53.3% (3 complete and 21 partial remissions) with a median remission duration of 9 months. Adeno- and squamous cell carcinoma responded to the chemotherapy with a remission rate of 63% (7 out of 11 patients) and 58% (14 out of 24 patients), but there were only 30% responders in large cell carcinoma (3 out of 10 patients). The median survival time for responders was 12 months, for those with no change and for patients with progressive disease 6 months. Myelosuppression and renal toxicity of the combination was generally not a treatment problem; subjective tolerance, however, (gastrointestinal upset) was poor. PMID- 3035449 TI - Immunological studies on herpetic keratitis: effect of IL-2 on HSV-specific CTL. AB - The effect of interleukin 2 (IL-2) on cytotoxic T lymphocytes (CTL) was examined in mice with corneal infection of herpes simplex virus (HSV). The corneas of the mice were inoculated with HSV after scratching the corneal epithelium. Ten days after the inoculation, CTL were induced in vitro from spleen cells of the mice. IL-2 was added for the initiation of CTL induction. CTL induced without IL-2 served as controls. After the culture, viable cells were counted and cytotoxicity of CTL was measured by 3H-proline releasing assay. IL-2 promoted proliferation of HSV specific CTL without increasing the cytotoxicity of HSV specific CTL. PMID- 3035450 TI - Cellular retinoic acid binding protein (CRABP) in retinoblastoma. AB - Binding proteins for retinoic acid (cellular retinoic acid binding protein, CRABP) have been demonstrated in various cell types, and display the characteristics of receptors. Three retinoblastomas are described. The clinical diagnosis of retinoblastoma was established by ophthalmoscopic echographic and histological examination. One part of the tumor was frozen in liquid nitrogen immediately after surgery and used for the determination of CRABP. CRABP was present in cells from all three tumors. This may indicate sensitivity of this tumor to retinoic acid or synthetic retinoic acid derivatives with biologic activity. PMID- 3035451 TI - Long-term results of proton beam irradiated uveal melanomas. AB - The first 128 consecutive patients with uveal melanomas treated with proton beam irradiation were studied in order to evaluate survival and visual acuity status of patients with relatively long-term follow-up. The median follow-up was 5.4 years, and no patient was lost to follow-up. All tumors showed regression. The most recent visual acuity was 20/40 or better in 35% and 20/100 or better in 58%. Eight eyes were enucleated because of complications. Metastasis developed in 26 patients (20.5%) from 3 months to 7 years after treatment. Results indicate that proton irradiation is quite successful for achieving local control of uveal melanomas. A large proportion of the treated eyes maintained useful vision. Five year follow-up data indicate that proton irradiation has no deleterious effect on the likelihood of the development of metastasis. PMID- 3035453 TI - Cytomegalovirus retinitis and response to therapy with ganciclovir. AB - A 15-month prospective study of 109 patients with the acquired immune deficiency syndrome (AIDS) or AIDS-related complex (ARC) was conducted. Cytomegalovirus (CMV) retinitis developed in 18 of these patients; they were treated with ganciclovir. Five other patients with CMV retinitis who were not part of the prospective study were also treated with ganciclovir. CMV retinitis frequently involved the peripheral retina. All 23 patients treated with ganciclovir showed clinical regression of retinitis, although breakthrough recurrence of CMV retinitis occurred in seven patients (30.4%) while on maintenance therapy with ganciclovir. During treatment, neutropenia (less than 1000 leukocytes/mm3) developed in three patients (13%). Ganciclovir is an effective means of therapy for CMV retinitis, but it must be given chronically to prevent reactivation. Breakthrough recurrences while on maintenance therapy are not uncommon, but can be successfully treated with more aggressive treatment with ganciclovir. In addition, the prognosis for survival of AIDS patients being treated with ganciclovir is improved when compared with that of untreated patients. PMID- 3035452 TI - Pathologic features of cytomegalovirus retinopathy after treatment with the antiviral agent ganciclovir. AB - Ganciclovir is a new antiviral compound (also called BW B759U, DHPG, BIOLF-62, and 2'NDG) that has been used for the treatment of cytomegalovirus (CMV) retinopathy in immunocompromised patients (bone marrow recipients or acquired immune deficiency syndrome [AIDS] victims). The authors studied the eyes of three AIDS patients with CMV retinopathy who died while receiving ganciclovir chemotherapy. Gross, microscopic, and ultrastructural studies of these cases showed varying degrees of retinal scarring and active CMV lesions at the margins of the scars. CMV antigens were localized in cells at all layers of retina at the border of the lesions and in isolated cells in a perivascular location within histologically normal appearing retina. These areas probably represent sites of recrudescence when the drug is discontinued. In situ hybridization using a cloned complementary DNA (cDNA) probe of human CMV corroborated the immunocytologic localization of the virus. Ultrastructural studies showed megalic syncytial cells containing mostly capsids exclusively in the cell nucleus. The cytoplasmic electron-dense membrane-bound bodies that have characterized untreated cases of CMV retinopathy were absent in the treated cases. An attempt to isolate CMV in tissue culture from the vitreous and retina of one of the cases yielded a negative result. Our results indicate that ganciclovir does not effectively eliminate CMV from the retina nor does it suppress expression of all viral genes. Ganciclovir appears to function by limiting viral DNA synthesis and subsequent packaging of viral DNA into infectious units, thereby acting as a virostatic chemotherapeutic agent. PMID- 3035454 TI - Intraoral neurilemmoma (schwannoma): an unusual palatal swelling. AB - A case of an uncommon intraoral tumor, the neurilemmoma, is reported. This tumor presented in an unusual way and exhibited clinical features that are classically associated with the pleomorphic adenoma. The subsequent histologic findings are presented, and an explanation for the clinical behavior of this lesion is suggested. The case for excision biopsy of palatal swellings is supported. PMID- 3035455 TI - Concomitant regional odontodysplasia and hydrocephalus. AB - The case of a patient with concomitant regional odontodysplasia, hydrocephalus, and mental retardation is presented. Tooth eruption was retarded, probably because of the presence of a cementum-like substance on the enamel surface as well as the presence of discontinuous odontogenic epithelium that surrounded the affected teeth. Neural damage during intrauterine life is suggested as a likely cause of the dental abnormalities. This hypothesis is supported by the fact that both primary and permanent teeth in the area were affected. It is further supported by the finding of dysplastic dentin. PMID- 3035456 TI - Salivary duct carcinoma: ultrastructural and histogenetic considerations. AB - Two cases of salivary duct carcinoma were examined by light and electron microscopy. Histologically, the tumor presented cribriform and papillary patterns together with comedonecrosis. Electron microscopy revealed duct cells with microvilli, interdigitations, and apical vesicles. No myoepithelial cells were observed. These findings suggest a ductal origin other than from the intercalated duct or its precursor element. The importance of separating the salivary gland adenocarcinomas is discussed. PMID- 3035458 TI - [Registration of patients with colonic polyposis in Hungary]. PMID- 3035457 TI - A histologic evaluation of a mandibular cross section one year after augmentation with hydroxyapatite particles. AB - A pathologic mandibular fracture in a 35-year-old woman 1 year after the attempted filling of a bone defect with particulate, dense hydroxyapatite resulted in a partial mandibular resection, allowing observation of cross sections of the mandible and associated soft tissue. Thin tissue sections were examined without decalcification. This permitted excellent observation of the hydroxyapatite particles, the bone, and the soft tissue. Osseointegration had occurred only in the areas closely associated with the bone. The immobility of the particles was a prerequisite for involvement with new bone formation. Hydroxyapatite particles in areas that allowed any mobility were surrounded by connective tissue with no bone formation evident. Heterogeneous particles were observed, indicating the possibility of a lack of purity in the hydroxyapatite ceramic, which may have contributed to the resorption of the particles. These findings, as well as other clinical and experimental findings, lead us to question the concept of hydroxyapatite as a bioactive ceramic that induces osteogenesis or osteoconductivity. PMID- 3035459 TI - Chemotherapy for salivary gland cancer. AB - Because of the scarcity of such lesions, little is known about the efficacy of chemotherapy for advanced salivary gland cancers. Although surgery and irradiation are the mainstays of treatment, patients with recurrent tumors and those with unresectable or metastatic cancer are not candidates for this usual approach. Ten patients with recurrent, metastatic, or unresectable salivary gland tumors were treated with combination chemotherapy, primarily with cisplatin, doxorubicin hydrochloride (Adriamycin), and cyclophosphamide, or cisplatin with 5 fluorouracil. In patients whose tumors exhibited no response, second-line drugs were used. The overall response rate was 50%--with one complete response--but the duration of response was short. This report contributes to the growing data base that demonstrates definite chemosensitivity of these tumors. To date, 116 patients have been reviewed. Adenocarcinoma-like cancers respond best to cisplatin, doxorubicin hydrochloride, and 5-fluorouracil. High-grade mucoepidermoid carcinoma may show a sensitivity similar to that of squamous cell carcinoma. Multi-institutional protocols need to be developed to assess the roles of adjuvant and palliative chemotherapy in the treatment of salivary gland cancer. PMID- 3035460 TI - Non-epidermoid carcinoma of the larynx: the Thomas Jefferson University experience. AB - Nonepidermoid cancers of the larynx appear with consistency in large series of laryngeal tumors. We reviewed 1135 laryngeal cancers seen over a 24-year period at Thomas Jefferson University Hospital and found a 1.7% incidence. This is similar to several other large series. These patients are presented in terms of their sex, age, race, clinical presentations, therapies, and outcomes, and a review of these unusual tumors is given. The recognition of such lesions is important because many require different therapies and have different prognoses than their squamous cell counterparts. The relatively infrequent occurrence of these tumors makes the availability of controlled clinical studies extremely difficult. PMID- 3035461 TI - Vertical nystagmus in routine caloric testing. AB - Vertical nystagmus may occur in caloric testing when only horizontal is expected. We examined this occurrence in 112 normal subjects and in 339 patients with dizziness. Vertical nystagmus was found in 29 percent of normals and in 12 percent of patients with dizziness, more often with hot than cool caloric stimuli and it is always accompanied by horizontal nystagmus. The finding occurred with peripheral and central nervous system diagnoses as well as with patients whose dizziness was considered psychogenic or was undiagnosed. Maximum slow component velocity (SCV) of vertical nystagmus was usually half or less than that of the nystagmus in the horizontal lead. The SCV time profiles of the nystagmus in horizontal and vertical leads differed considerably. Possible origins of vertical nystagmus are discussed. Whatever the origin, it is clear that the finding of vertical nystagmus in routine caloric testing does not automatically denote disease of the central nervous system. PMID- 3035462 TI - Malignant fibrous histiocytoma. PMID- 3035463 TI - Short latency somatosensory evoked potentials in patients with painful dysaesthesias in peripheral nerve lesions. AB - Abnormally delayed and attenuated short latency somatosensory evoked potentials after electrical stimulation in the distribution of painful dysaesthesias after peripheral nerve lesions are reported in 4 selected patients with lesions in very proximal parts of the peripheral nerves. These included painful spondylopathic cervical radiculopathy, meralgia paraesthetica, pyriformis syndrome and allodynia after injury of the femoral nerve. Locally developed techniques for testing musculocutaneous, lateral femoral cutaneous and saphenous nerves were used in 3 patients. Possible pathophysiological mechanisms causing evoked potential abnormalities are discussed. PMID- 3035464 TI - Genomic cloning of human Echinococcus granulosus DNA: isolation of recombinant plasmids and their use as genetic markers in strain characterization. AB - A small, size selected (0.5-5.0 Kbp) genomic DNA library has been constructed in the bacterial plasmid pAT153 using DNA extracted from a human isolate (Kenyan origin) of the hydatid disease organism, Echinococcus granulosus. A panel of taeniid cestode DNAs has been used in conjunction with hybridization and restriction-enzyme analysis to identify in the library two recombinant plasmids with Echinococcus-specific inserts and a single recombinant plasmid (coded pEG18) with a DNA fragment unique for E. granulosus. These, and other recombinant plasmids with E. granulosus DNA inserts, have been used in restriction endonuclease, Southern transfer and hybridization analysis to independently and reproducibly discriminate between the UK horse and sheep strains of E. granulosus; these probes may well prove of value for characterizing isolates from other endemic areas. The feasibility of using a cloned DNA fragment as the basis of a simple field test for distinguishing eggs of E. granulosus from those of other taeniid cestodes is discussed. The recombinant insert (approximately 2.3 Kbp in size) of pEG18 has a low copy number - estimated at approximately 26 - and it may not be sufficiently sensitive for practical use as a DNA probe in the identification of small numbers of E. granulosus eggs by molecular hybridization. PMID- 3035465 TI - [DNA-topoisomerase II: Cellular targets of antineoplastic agents]. PMID- 3035466 TI - [Coxsackie B virus infections in cardiology. Apropos of 66 cases]. AB - Coxsackie B virus infections are common and frequently asymptomatic. However in young people, they can cause primary congestive cardiomyopathy and complicate previous cardiovascular illnesses. Virus diagnosis is difficult and based mainly on the detection of significant rising or stating high neutralizing antibody titers. A clinical and epidemiological five years study investigated 3,856 sera. 30.2% of the patients had evidence of a significant antibody titer (greater than or equal to 64) to one of the group B Coxsackie viruses; this percentage reaches 34.6% in cardiology and fluctuates from year to year; Coxsackie B2 is predominant (55.9%) in cardiology; coxsackie B1 and B2 antibody responses were detected more frequently than B3, B4 and B5. Coxsackie B6 appears to be uncommon. From these, 66 patients have evidence of coxsackie B virus infections with 60.2% for Coxsackie B2; they include pericarditis (30.3%), acute and chronic congestive cardiomyopathies (19.7%). Moreover it has been suggested that Coxsackie B virus might be responsible for electrocardiographic abnormalities (9.1%), ischemic heart disease and myocardial infarction. Enzyme linked immunosorbent assay (ELISA) and radioimmunoassay with purified antigens (VP1 and VP4) did not appear better than microneutralisation for evaluation of IgM and IgG antibodies. To elucidate the mechanisms of cardiac injury it is refer to viral replication, virus specific and auto-reactive T cells cytotoxic activities. PMID- 3035467 TI - [Pulmonary emphysema induced by elastase: influence of the dose and effect of added collagenase]. AB - The endotracheal deposition of pancreatic porcine elastase (EPP) at 40 u X kg-1 body weight provokes a typical pulmonary emphysema (alveoli disruption) in rats. This emphysema is significant (p less than 0.001) when quantitatively compared to a placebo (100% increase of the mean linear intercept, ILM). The EPP-treated lungs are very heterogeneous and the size of the alveoli vary as much as 30% versus 19% in controls. The emphysema is more effective at a 80 u X kg-1 dose and the individual disparities are reduced (dose effect). When bacterial collagenase (200 u X kg-1 body weight) is added to EPP, the pulmonary abnormalities (disruptions, ILM) are in no way increased (no enzymatic synergy, repair by collagenesis?). In contrast, alveolar dilation is slightly reduced 8 weeks after enzyme administration (p less than 0.05): EPP is not altered in vitro by collagenase and despite several hypothesis, the moderating effect of collagenase in vivo still remains unexplained. This result suggests that the joint presence of several proteases is not necessarily an aggravating factor in the etiopathogenesis of emphysema. PMID- 3035468 TI - [Pharmacological bases of the treatment of cardiac insufficiency]. AB - Congestive heart failure is a complex physiopathological state where both myocardial hypo-contraction and excessive peripheral vasoconstriction lead to lower cardiac output. The increase in cytosolic calcium concentration triggers the contractile processus. Digitalis inhibits the Na+/K+ ATPase enzyme and indirectly increases intracellular calcium concentration. beta 1 agonists increase the synthesis of cAMP-dependent protein kinase and hence the recruitment of new receptor-operated calcium channels which increase the calcium influx and the mobilization from its intracellular storage sites. Vascular smooth muscle contraction occurs with calcium influx into the cell resulting from various receptor activation. In congestive heart failure, activation of the sympathetic nervous system and of the renin-angiotensin system leads to neurohumoral-induced peripheral vasoconstriction. Renal effects of angiotensin II and aldosterone are responsible for sodium and water retention. alpha 1-blocking agents are drugs that block competitively the catecholamines effects on vascular receptors. Angiotensin I-converting-enzyme inhibitors block the formation of the key-element of the system: angiotensin II. Both alpha 1-blocking agents and converting-enzyme inhibitors show vasodilatator effects and acutely improve hemodynamic status of patients with congestive heart failure. Converting-enzyme inhibitors exhibit specific improvement of intrarenal hemodynamics and do not induced sodium and water retention in longterm therapy. PMID- 3035469 TI - Preservation of renal structure and function in the rat remnant kidney model of chronic renal failure by enalapril treatment. AB - The remnant kidney model of chronic renal failure was established in rats subject to subtotal (1 7/8) nephrectomy and the evolution of renal injury studied over a period of 6 wk. One wk after subtotal nephrectomy, rats had a mean conscious systolic blood pressure of 158 +/- 5 mm Hg and serum creatinine of 128 +/- 9 mumol/l. Both systolic blood pressure and serum creatinine rose over the next 5 wk in concert with progressive glomerulosclerosis and proteinuria. Enalapril, an angiotensin converting enzyme inhibitor, was administered (5 mg/kg/day) to rats (n = 11) from 1 wk after subtotal nephrectomy. Enalapril lowered systolic blood pressure over the treatment period. Systolic blood pressure was 122 +/- 5 mm Hg compared with 176 +/- 7 mm Hg in untreated rats (p less than 0.001) at 6 wk. Serum creatinine 6 wk after subtotal nephrectomy was 110 +/- 9 mumol/l with enalapril treatment, compared with 159 +/- 21 mumol/l (p less than 0.025) in control animals. Enalapril treated rats had lower urinary protein excretion than controls (15 +/- 3 mg/24 hr vs 85 +/- 22 mg/24 hr, p less than 0.0001) at 6 weeks. Glomerulosclerosis, assessed by blinded histological score, was also reduced in the enalapril treated group (1.79 +/- 0.08 vs 2.36 +/- 0.16, p less than 0.01). Enalapril treatment was associated with a reduction in filtration fraction (51Cr-EDTA/125I-hippurate clearance). At 6 wk, filtration fraction was 0.30 +/- 0.03 in enalapril treated and 0.48 +/- 0.03 in control rats (p less than 0.001). Enalapril treatment in the subtotal nephrectomy model of renal failure preserved renal structure and function.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3035470 TI - Assessment of precancerous lesions of the uterine cervix for evidence of human papillomavirus infection: a histological and immunohistochemical study. AB - Cervical biopsies obtained by colposcopic direction from 358 women were histologically examined for squamous dysplasia (cervical intra-epithelial neoplasia; CIN) and human papillomavirus (HPV) infection. Of the 358 biopsies, 136 were stained by an immunoperoxidase method using an antiserum against genus specific (common) antigen of bovine papillomavirus. HPV antigens were detected in 40% of biopsies showing definite histological evidence of HPV effect, and in 7.9% and 2.6% of those with possible or no HPV effect, respectively. HPV effect was commonly seen in association with CIN. The frequency of histological evidence of HPV effect and positive immunoperoxidase staining decreased with increasing grades of CIN. HPV antigen was found in 57% of areas of HPV change with minor atypia, 34% of zones of CIN I and in only 8% of zones of CIN II. No antigenic staining or definite histological evidence of HPV effect was observed within areas of CIN III. Antigen was generally confined to the nuclei of superficial koilocytes, cells with lesser degrees of perinuclear clearing and parakeratotic cells. These results how a strong association between HPV infection and precancerous lesions of the uterine cervix and are consistent with the hypothesis that production of the HPV structural antigen requires a high degree of squamous cell maturation. The immunoperoxidase findings and the histopathological observations support the view that HPV change and dysplasia are part of a morphological continuum in which the cytopathic effect of HPV is expressed mainly in lower grades of dysplasia. PMID- 3035471 TI - Recurrent dedifferentiated liposarcoma of the spermatic cord simulating malignant fibrous histiocytoma: an immunohistochemical and ultrastructural study. AB - A case of recurrent dedifferentiated liposarcoma simulating malignant fibrous histiocytoma, with complete absence of lipoblastic differentiation, is described. The tumour cells showed strongly positive immunostaining for alpha-1 antichymotrypsin. Electron microscopy revealed a mixture of fibroblasts and histiocytes. Our findings suggest that the dedifferentiated component reflects an altered differentiation pathway of the primitive mesenchymal cells in the original liposarcoma. PMID- 3035472 TI - Specific 1,25-hydroxycholecalciferol receptors and stimulation of 25 hydroxycholecalciferol-24R hydroxylase in human amniotic cells. AB - We have analyzed the 1 alpha, 25-dihydroxycholecalciferol [1,25(OH)2D3] receptor content of cultured cells from human amniotic fluid. Six cell lines were grown to confluence in a minimum essential medium containing 20% fetal calf serum. All had a normal karyotype, five were male and one was female. Hypertonic cytosol extracts were prepared by sonication followed by centrifugation at 200,000 X g 30 min. Saturation analysis was performed by incubating the extracts with [3H] 1,25(OH)2D3 (20-500 pM, 160 Ci/mmol) with and without 100-fold molar excess of unlabeled 1,25(OH)2D3. Linear sucrose gradient (5-20% w/v) analysis was performed with 1.5 nM [3H]-1,25(OH)2D3 alone or in presence of 100-fold molar excess, 1,25(OH)2D3. Functional responsiveness was measured by induction of 25 hydroxycholecalciferol-24R-hydroxylase with 1 and 10 nM 1,25(OH)2D3. The six cell lines studied had receptors with dissociation constant of 44 +/- 6 pM (mean +/- SEM). The binding capacity was 10,200 +/- 1,750 sites/ng protein (mean +/- SEM) with extreme values of 4,700 and 15,500. A single peak for specific binding migrating at approximately 3S was observed by sucrose gradient centrifugation. 25 Hydroxycholecalciferol-24R-hydroxylase was induced by 1 and 10 nM 1,25(OH)2D3 in a dose-dependent fashion. The data show that receptors for 1,25(OH)2D3 are present in cultured amniotic fibroblast-like cells early in pregnancy. These cells may thus prove to be useful for further characterization of 1,25(OH)2D3 receptors in fetal tissue. PMID- 3035473 TI - [Clinical picture of Coxsackie B virus myocarditis in children]. PMID- 3035474 TI - [A case of myocardial infarction in an 11-year-old girl with myocarditis]. PMID- 3035475 TI - Cytomegalovirus infection in an infant presenting with cutaneous vasculitis. PMID- 3035476 TI - Acquired immunodeficiency syndrome in children: report of the Centers for Disease Control National Surveillance, 1982 to 1985. AB - Since national surveillance for acquired immunodeficiency syndrome (AIDS) began in 1981, the Centers for Disease Control (CDC) has received reports of more than 20,000 cases of AIDS in the United States. As of December 31, 1985, 307 of these cases had been diagnosed in children younger than 13 years of age. The number of cases is increasing rapidly. The number of cases reported in 1985 more than doubled those reported in 1984. The major risk factors in children for acquiring infection with the causative agent, human immunodeficiency virus (HIV), were having a mother known to be infected and/or at increased risk for infection and receiving a transfusion of blood or blood products. Of the 307 children with AIDS, 73% were reported from one of four states: New York, New Jersey, Florida, and California. Most AIDS cases in children occur in black or Hispanic infants and toddlers. The estimated incubation period for AIDS in children has increased each surveillance year, with the longest incubation exceeding 7 years. The prognosis for children with AIDS is poor and infants less than 1 year of age have the shortest survival time following diagnosis. Continued national surveillance for AIDS is mandatory for establishing effective prevention programs to control the spread of the disease. The CDC encourages all health care personnel to report cases of AIDS to their public health departments. PMID- 3035477 TI - Infantile spasms due to unilateral cerebral infarcts. PMID- 3035480 TI - Indirect immunofluorescence and generation of cyclic-AMP investigation with Graves' patients' sera. AB - Generation of cAMP was measured on human thyroid slices and indirect immunofluorescence was carried out on frozen human thyroid sections. Sera from 16 previously not treated thyrotoxic patients and from 5 euthyroid patients were used. The majority of the thyrotoxic sera (13 out of 16) provoked enhanced generation of cAMP and the same cases showed positivity by indirect immunofluorescence. The possible connection between enhanced generation of cAMP and detectability of antibodies binding to different thyroid structures is discussed. PMID- 3035478 TI - Varicella-like illness caused by live varicella vaccine in children with acute lymphocytic leukemia. AB - A varicella-like illness occurred in five of 52 children with acute lymphocytic leukemia following the administration of live varicella vaccine. Only one of the children required treatment with acyclovir. Virus isolated from two of the children was "vaccine-like" but differed slightly from the original vaccine strain when tested by restriction enzyme analysis. There did not appear to be a reversion to virulence because two of the household contacts who seroconverted had mild or subclinical infections. Vaccinees in whom this reaction developed tended to have a poor cellular immune response to varicella-zoster virus. PMID- 3035481 TI - Decrease in the response to ADH of the rat kidney as a result of early postnatal treatment with cortisol. AB - Wistar rats were injected just once, intraperitoneally with cortisol (1 microgram/g) or saline at the age of 5 days. The cortisol-treated rats did not differ significantly in the (U/P)osm ratio from the saline-treated controls before 15 days of life. Their response to ADH was distinct but weaker than in the saline controls aged 30 days. This reduced response persisted to 60 days of life. In the collecting tubule fragments, (3H)AVP specific binding was lower in the cortisol-treated rats than in the controls at the age of 20 and 60 days. There was no (3H)AVP specific binding in the proximal convoluted tubules in the cortisol- and saline-injected rats of both ages. The ontogenetic patterns of cAMP specific binding in the papillary cytosolic fraction were different: the early increase in cAMP binding was protracted in the cortisol-treated rats, and no peak appeared at the age of 25 days. Cytosolic protein kinase activity was lower, no peak appeared at 30 days, no activation of protein kinase occurred to the end of weaning in the cortisol-treated rats. The difference between the cortisol and saline groups was abolished by day 30. The interference of cortisol with the ontogenetic changes in AVP binding capacity and cAMP-dependent protein kinase appears to be a plausible cause of the altered development of the response to ADH. PMID- 3035479 TI - Nabilone versus prochlorperazine for control of cancer chemotherapy-induced emesis in children: a double-blind, crossover trial. AB - In a randomized, double-blind, crossover trial, nabilone was compared to prochlorperazine for control of cancer chemotherapy-induced emesis in 30 children 3.5 to 17.8 years of age. All subjects received two consecutive identical cycles of chemotherapy with the trial antiemetics given in accordance to a body weight based dosage schedule beginning eight to 12 hours before treatment. The overall rate of improvement of retching and emesis was 70% during the nabilone and 30% during the prochlorperazine treatment cycles (P = .003, chi 2 test). On completion of the trial, 66% of the children stated that they preferred nabilone, 17% preferred prochlorperazine, and 17% had no preference (P = .015, chi 2 test). Major side effects (dizziness, drowsiness, and mood alteration) were more common (11% v 3%) during the nabilone treatment cycles. CNS side effects appeared to be dose related and were most likely to occur when the nabilone dosage exceeded 60 micrograms/kg/d, but individual tolerance to nabilone varied considerably. Lower dosages of nabilone were associated with equivalent efficacy and no major side effects. Nabilone appears to be a safe, effective, and well-tolerated antiemetic drug for children receiving cancer chemotherapy. Although major side effects may occur at higher dosages, nabilone is preferable to prochlorperazine because of improved efficacy. PMID- 3035484 TI - [Effect of whole body irradiation on O2- production in polymorphonuclear leukocyte in guinea pig]. PMID- 3035483 TI - Calmodulin-like proteins and communicating junctions. Electrical uncoupling of crayfish septate axons is inhibited by the calmodulin inhibitor W7 and is not affected by cyclic nucleotides. AB - The effects of W7, a calmodulin (CaM)-inhibitor, and cyclic nucleotides on electrical coupling and uncoupling are studied in crayfish lateral giant axons (septate axons). The septate axons provide a relatively simple two cell system in which both surface membrane and junctional resistance can be measured independently. Four microelectrodes are inserted into a septate axon, two on each side of the septum. Hyperpolarizing current pulses (150 nA) are injected alternatively in the caudal and rostral axon segment and the resulting electrotonic potentials are recorded. The axons are uncoupled at regular intervals by superfusing them with acetate-containing saline solution (pH 6.3) in the presence or absence of W7 (50-100 microM) or, as a control, its nonchlorinated form (W5). W7 strongly inhibits the acetate-induced increased in junctional resistance, while W5 is ineffective. The uncoupling inhibition does not appear to be caused by an increase in cyclic nucleotide concentration, because in preliminary experiments exposure to db-cAMP or db-cGMP (up to 1 mM) does not seem to influence either the basic values of Rj or their changes with acetate. The data confirm previous evidence for a participation of CaM-like proteins in cell-to-cell channel gating. PMID- 3035482 TI - Atrial natriuretic peptide receptors along the rat and rabbit nephrons: [125I] alpha-rat atrial natriuretic peptide binding in microdissected glomeruli and tubules. AB - Binding of [125I] alpha-rat atrial natriuretic peptide ([125I] alpha-RANP) was measured in glomeruli and pieces of tubule microdissected from rat and rabbit nephrons. High densities of specific ANP binding sites were found only in the glomeruli (10-30 X 10(-18) mol X glom-1), whereas no specific binding could be detected in the proximal tubule, the thin segments of the Henle's loop, the thick ascending limb, the distal tubule and the cortical and outer medullary collecting tubules. Rising the temperature from 4 degrees C to 35 degrees C resulted in biphasic kinetics of binding, suggesting a temperature-dependent inactivation of labelled hormone by glomeruli. At 4 degrees C, specific binding of [125I] alpha RANP was time and dose-dependent and Scatchard analysis of data indicated an apparent equilibrium dissociation constant of 0.63 nM. Competition experiments revealed the following sequence of stereospecificity for binding to rat glomeruli: RANP 3-28 greater than [125I] alpha-RANP = [125I] alpha-HANP = alpha RANP = antriopeptin III greater than antriopeptin II, whereas binding was unaffected by pharmacological doses of unrelated peptide hormones, prostaglandins, adrenergic agonists, dopamine, histamine and carbamylcholine. The results indicate that glomerular binding sites might be the physiological ANP receptors. PMID- 3035485 TI - [Prevention and treatment of cytomegalovirus infections after graft of allogenic bone marrow]. AB - Cytomegalovirus (CMV) infection is the most frequent cause of lethal infection after bone marrow transplantation. Viremia occurs in 50% of patients seropositive for CMV before transplantation. Interstitial pneumonitis due to CMV occurs in 10% to 20% of patients with 85% mortality. It is known that CMV infection is due to host reactivation of latent CMV infection or to the transmission of the virus by the marrow donor or by blood transfusions. Treatment of CMV infection has been disappointing in the past. All attempts to treat CMV pneumonia with available agents have failed. Recent studies have indicated the usefulness of prophylactic measures and the early treatment of CMV infections. The use of hyperimmune gammaglobulins has given contradictory results. The selection of seronegative marrow donors or blood donors is useful only if the recipient is seronegative. New antiviral drugs have been used recently in preliminary clinical trials. In preliminary studies a guanosine analogue similar to Acyclovir (DHPG Synthex or BWB 759 U Wellcome) has given reasonable hope of disease cure if it is used early before the occurrence of pneumonia. Phosphonoformate (Foscarnet) has also been shown to be active against CMV infection. Both drugs have good antiviral and clinical action in immunosuppressed patients but the results have been disappointing in cases of pneumonia. Relapse occurs frequently after cessation of the treatment and attempts are being made to use maintenance therapy. PMID- 3035486 TI - Nucleotide sequence analysis of the L gene of Newcastle disease virus: homologies with Sendai and vesicular stomatitis viruses. AB - The nucleotide sequence of the L gene of the Beaudette C strain of Newcastle disease virus (NDV) has been determined. The L gene is 6704 nucleotides long and encodes a protein of 2204 amino acids with a calculated molecular weight of 248822. Mung bean nuclease mapping of the 5' terminus of the L gene mRNA indicates that the transcription of the L gene is initiated 11 nucleotides upstream of the translational start site. Comparison with the amino acid sequences of the L genes of Sendai virus and vesicular stomatitis virus (VSV) suggests that there are several regions of homology between the sequences. These data provide further evidence for an evolutionary relationship between the Paramyxoviridae and the Rhabdoviridae. A non-coding sequence of 46 nucleotides downstream of the presumed polyadenylation site of the L gene may be part of a negative strand leader RNA. PMID- 3035488 TI - A new repetitive element of the CR1 family downstream of the chicken vitellogenin gene. AB - We have analyzed a repetitive DNA sequence found in the 3'-flanking region of the chicken vitellogenin gene. By its sequence, the repetitive DNA has been identified as a hitherto unreported member of the chicken CR1 family of repetitive elements. The CR1 sequence displays the structural characteristics of a long terminal repeat located at the 3' end of an avian retrovirus. The CR1 element lies 2.2 kb downstream of the vitellogenin gene and 'points' away from the gene rather than toward it. In this respect, this element differs from other CR1 repeats. The CR1 element is embedded in a region showing changes in chromatin structure implying a potential role for this sequence in determining the structural state of the local chromatin. PMID- 3035490 TI - Nucleotide sequence of a human ubiquitin Ub B processed pseudogene. PMID- 3035487 TI - The L1 family (KpnI family) sequence near the 3' end of human beta-globin gene may have been derived from an active L1 sequence. AB - We previously reported that some L1 family (KpnI family) members are closely associated with the Alu family sequence. To understand the details of the L1-Alu association, the structure of a L1-Alu unit downstream from the beta-globin gene was compared between human and primates. The results revealed that the L1-Alu associated sequence was formed by the insertion of the L1 sequence, T beta G41, into the 3' poly A tract of the preexisting Alu family sequence. It was estimated that the T beta G41 sequence was inserted after the divergence of Old World monkeys and hominoids and before the divergence of orang-utan and common ancestor of other higher hominoids. From the calculation of the mutation rates of L1 sequences, it was suggested that the T beta G41 was derived from an active L1 sequence which was able to encode reverse transcriptase-related protein. PMID- 3035491 TI - Regulatory sequences of endogenous mouse mammary tumor virus locus Mtv-8 from different mouse strains. PMID- 3035492 TI - The complete nucleotide sequence of bovine rotavirus C486 gene 4 cDNA. PMID- 3035489 TI - Competition studies with repressors and activators of viral enhancer function in F9 mouse embryonal carcinoma cells. AB - DNA competition studies have been used to investigate the presence of a repressor of viral enhancer function in F9 mouse embryonal carcinoma cells. The complete polyoma virus enhancer region, cotransfected into F9 cells with the SV40 promoter/enhancer attached to a chloramphenicol acetyl transferase marker gene, induced a small increase in pSV2CAT expression. This can be explained by preferential but weak binding by polyoma sequences of a molecule repressing pSV2CAT transcription. Repressor activity substantially disappeared when the cells were induced to differentiate by retinoic acid. Repressor binding was localised to one half of the polyoma enhancer, but was lost on further fragmentation of this region. It appears that multiple sequence elements may be required for repressor binding and that these are at least partially separable from the complement of elements binding enhancer activating molecules. PMID- 3035495 TI - Yeast regulatory sequences preferentially adopt a non-B conformation in supercoiled DNA. AB - Mung bean nuclease was used to probe for DNA unwinding in torsionally-stressed chimeric plasmids containing two micron plasmid sequences and the yeast LEU2 gene in a pBR322 vector. The yeast sequences are cleaved at only two sites, both of which map to regulatory regions: (1) the autonomously replicating sequence (ARS), an origin of DNA replication, of the two micron plasmid and (2) the transcription terminator region of the LEU2 gene. Nucleotide level analysis of the nuclease cleavage pattern shows that an A + T-rich structure, distinct from other non-B DNA conformations, is recognized. A computer analysis reveals that A + T content alone is not sufficient to explain the preferential occurrence of the A + T-rich structure in the ARS over other sequences of equal A + T content. The A + T-rich structure detected in the ARS maps to sequences required for DNA replication. Our findings demonstrate the DNA conformational flexibility of certain yeast regulatory regions and provide support for the hypothesis that the A + T-rich sequence in the ARS plays a role in DNA unwinding during the initiation of DNA replication. PMID- 3035494 TI - Cosmid mapping of the human chorionic gonadotropin beta subunit genes by field inversion gel electrophoresis. AB - A cosmid clone containing the entire hCG beta gene cluster has been isolated. The restriction map of this clone has been determined by an indirect-end-label FIGE (field inversion gel electrophoresis) method. Analysis of this cosmid clone shows that there are 6 hCG beta genes in human genomic DNA. A previously uncloned portion of the hCG beta cluster, termed the "gap" region, has been shown not to contain any sequences homologous to the hCG beta cDNA. The restriction mapping method employed in this study takes advantage of the superior resolution of FIGE for high molecular weight DNA fragments in the size range 15-50 kb. This method is broadly applicable and permits rapid and accurate restriction mapping for extended regions of genomic DNA that have been cloned into cosmid or lambda vectors. PMID- 3035497 TI - Clustered somatic mutations in and around first exon of non-rearranged c-myc in Burkitt lymphoma with t(8;22) translocation. AB - We have examined the restriction map of the c-myc gene in 15 BL cell lines carrying the variant t(8;22) translocation in which c-myc is known to remain on chromosome 8. Using 3 restriction enzymes cutting outside the c-myc domain (EcoRI, BamHI, HindIII), we found no evidence for a c-myc/Ig lambda rearrangement in 14 BL cell lines. In the last one, BL 37, the 3' flanking region was rearranged corresponding to the already identified breakpoint located 400 pb downstream from the c-myc gene (9). Using 4 restriction enzymes cutting inside the c-myc gene (PvuII, PstI, SacI, HincII) we looked for discrete abnormalities within the gene limits, and we found in 9 BL cell lines several abolished and created sites, compatible with multiple independent somatic mutations. They are significantly clustered in the 5' non coding region, with a striking prevalence at the end of exon 1. The role of mutations in the non-coding first exon region for the deregulation of c-myc expression is discussed. PMID- 3035496 TI - Activities of herpes simplex virus type 1 (HSV-1) ICP4 genes specifying nonsense peptides. AB - Synthetic oligonucleotide linkers containing translational termination codons in all possible reading frames were inserted at various positions in the cloned gene encoding the herpes simplex virus type 1 (HSV-1) immediate-early regulatory protein, ICP4. It was determined that the amino-terminal 60 percent of the ICP4 gene was sufficient for trans-induction of a thymidine kinase promoter-CAT chimera (pTKCAT) and negative regulation of an ICP4 promoter-CAT chimera (pIE3CAT); however, it was relatively inefficient in complementing an ICP4 deletion mutant. The amino-terminal ninety amino acids do not appear to be required for infectivity as reflected by the replication competence of a mutant virus containing a linker insertion at amino acid 12. The size of the ICP4 molecule expressed from the mutant virus was consistent with translational restart at the next methionine codon corresponding to amino acid 90 of the deduced ICP4 amino acid sequence. PMID- 3035493 TI - RNA polymerase II terminates transcription in vitro in the SV40 origin region. AB - To begin to study signals on DNA that can cause mammalian RNA polymerase II to terminate transcription, we examined the RNA transcripts produced from a number of different DNA templates in a HeLa whole-cell extract. When transcripts initiating from the strong adenovirus late promoter and extending through the SV40 promoter-replication origin region were analyzed, it was observed that a significant fraction (approximately 75%) were terminated within these SV40 sequences. These transcripts, the 3' ends of which were mapped to several sites within an approximately 200 base pair region, appeared to result from transcription termination, as judged by kinetic and pulse-chase experiments. The possible significance of these findings with respect to SV40 gene expression in particular and transcription termination in general are discussed. PMID- 3035499 TI - The nucleotide sequence of the 5' and 3' ends of rotavirus SA11 gene 4. PMID- 3035498 TI - 1H NMR study of the exchangeable protons of the duplex d(GCGTTGCG).d(CGCAACGC) containing a thymine photodimer. AB - A comparison is presented of the imino proton NMR spectra of the double stranded octamer d(GCGTTGCG).d(CGCAACGC) and the same octamer in which the two central thymine residues occur as a cis-syn thymine dimer. Except for the terminal base pairs all imino protons were detected and assigned in the NMR spectrum. The spectra show that in the thymine dimer duplex, contrary to common belief, all base pairs occur in a hydrogen bonded form, although the hydrogen bonds of the two central AT base pairs are substantially weakened. The melting temperature decreases about 13 degrees C on thymine dimer formation. PMID- 3035500 TI - The flexibility and topology of simian virus 40 DNA in minichromosomes. AB - The linking number of DNA in minichromosomes increases by 2 turns during SV40 assembly. Changes in temperature also influence the average linking number of the total intracellular forms of SV40 DNA. When the isolated minichromosomes assembled in vivo are incubated with topoisomerase I at 33 degrees C in vitro, the linking number of SV40 DNA decreases. This decrease is about: -1.1 turns for minichromosomes with an average nucleosome spacing of 198 base pairs (bp), wt776; and -0.6 turns for minichromosomes containing a shorter average nucleosome repeat (177 bp), tsC219. The difference between the average linking number of naked SV40 DNA relaxed with topoI at 33 degrees C and minichromosomes relaxed with the enzyme at the same temperature indicates that SV40 chromatin contains on the average 26 nucleosomes. However, the results of studies obtained both on DNA flexibility in chromatin and in naked DNA, and on the shape of the topoisomer distribution curves, indicate that all of the minichromosomes, regardless of their overall structure, do not contain the same number of nucleosomes; this heterogeneity may be as large as 8 nucleosomes. We find no apparent correlation between the amount of minichromosomes containing unconstrained torsional stress and the abundance of the molecules with a structure characteristic of transcriptionally active chromatin. PMID- 3035503 TI - Two restriction endonucleases from Bacillus sphaericus: BspXI and BspXII. PMID- 3035504 TI - Sequence correction and identification of a Rsa I site 3' to human G gamma globin gene. PMID- 3035505 TI - Nucleotide sequence of the homoserine kinase (thr B) gene of Brevibacterium lactofermentum. PMID- 3035501 TI - Dual tissue-specific expression of apo-AII is directed by an upstream enhancer. AB - Apolipoprotein-AII (apo-AII) is one of a family of evolutionarily related proteins which play a crucial role in lipid transport and metabolism. The serum levels of human apo-AII have been shown to be inversely correlated to the incidence of coronary heart disease and its expression to be limited to the liver and intestine. Here we demonstrate that this dual tissue-specificity involves DNA sequences located in a 259 bp region centred 782 bp upstream from the transcription initiation site. These sequences function in an orientation independent manner and are absolutely required for transcription from the apo-AII promoter. The regulatory region contains sequences which are homologous to the apo-AI, beta-globin and immunoglobulin gene promoters and to the immunoglobulin heavy-chain enhancer. PMID- 3035502 TI - Structure and sequence of a UDP glucose pyrophosphorylase gene of Dictyostelium discoideum. AB - Cell-cell contact and exogenous cAMP regulate the expression of uridine diphosphoglucose pyrophosphorylase (UDPGP) of Dictyostelium discoideum (B. Haribabu, A. Rajkovic and R. P. Dottin, 1986, Dev. Biol., Vol. 113, 436-442). cAMP appears to regulate gene expression in Dictyostelium by transmembrane signal transduction (B. Haribabu and R. Dottin, 1986, Mol. Cell. Biol. 6, 2402-2408). To further characterize the mechanism of action of cAMP on the expression of this gene and the nature of the defects in UDPGP mutants that abort development, we sequenced the cDNA and the genomic DNA, including intervening and flanking sequences. The deduced amino acid sequence predicts a polypeptide of 57,893 d. molecular weight. Three short (100-200 nucleotides) A+T rich introns occur within the coding sequences but only one of them contains a sequence TAACTAAC, similar to the yeast lariat acceptor site. The 5' flanking sequences are also A+T rich and contain an oligo A tract (-14 to -24), a TATA box (-25 to -32), and a short G+C rich region (-63 to -101) which may be a control region. From -196 to -209 is a sequence AAAGTAGTATTCAA which matches in 11 of its 14 nucleotides, a sequence found upstream from the hormonally regulated P-enolypyruvate carboxykinase gene of rat. PMID- 3035506 TI - Nucleotide sequence of a Bacillus megaterium gene homologous to the dnaK gene of Escherichia coli. PMID- 3035507 TI - Pyrophosphate scintigraphy and other non-invasive methods in the detection of cardiac involvement in some systemic connective tissue diseases. AB - Thirteen patients with systemic lupus erythematosus, 8 patients with polymyositis, and 6 patients with spondylitis ankylopoetica (Bechterew's disease) underwent clinical cardiologic examination and scintigraphy of the myocardium (99mTc-pyrophosphate), ECG, echocardiography, polygraphy, and their blood pressure was taken. The aim of the study was to ascertain how such a combination of non-invasive examinations can help in recognizing a cardiac involvement. In systemic lupus erythematosus cases one or more positive findings were revealed in 9 patients (69%), in 4 patients all examinations were negative (31%). Four patients (50%) with polymyositis had positive findings. In patients with spondylitis ankylopoetica positive findings occurred in 2 cases (33%). The study has shown that a combination of non-invasive cardiologic methods increases the probability of detecting cardiac involvement in systemic connective tissue diseases. PMID- 3035509 TI - [Clinical implications of biological differentiation of small cell carcinoma of the lung]. PMID- 3035508 TI - [Prevalence of rotaviruses, Campylobacter and pathogenic enterobacteria in feces of subjects residing in the Urbino area]. PMID- 3035510 TI - [Interstitial pneumonia as the cause of death during treatment of small-cell lung cancer]. PMID- 3035511 TI - [Polychemotherapy using methotrexate, adriamycin, cyclophosphamide and CCNU (MACC) in patients with advanced adenocarcinoma and large cell carcinoma of the lung]. PMID- 3035512 TI - [Effect of intravenous hydrocortisone and intramuscular ACTH on spirometric indicators in patients with partially reversible bronchial stenosis]. PMID- 3035513 TI - Proliferation of thyrotropin releasing hormone receptors in specific brain regions during the development of hypertension in spontaneously hypertensive rats. AB - The binding of [3H] [3-MeHis2] thyrotropin releasing hormone [( 3H]MeTRH) to brain membranes prepared from 8 week old spontaneously hypertensive (SHR) and normotensive Wistar-Kyoto (WKY) rats was determined. [3H]MeTRH bound specifically to rat brain membranes at a single high affinity site. The density (Bmax value) of [3H]MeTRH binding sites was significantly greater (28%) in SHR rats compared to WKY rats. The apparent dissociation constants (Kd values) for the binding of [3H]MeTRH in SHR and WKY rats did not differ. Binding in the various brain regions revealed that the density of [3H]MeTRH was highest in the hypothalamus followed in decreasing order by pons + medulla, midbrain, cortex and striatum. The binding of [3H]MeTRH was approximately 25% greater in cortex, hypothalamus and striatum of SHR rats in comparison to WKY rats. The binding in pons + medulla, midbrain and pituitary of SHR and WKY rats did not differ. To assess the significance of increased binding sites for [3H]MeTRH in some brain regions of SHR rats, the binding studies were carried out during normotensive and hypertensive stages of postnatal age in the two strains. In 3 and 4 week old SHR rats there was neither an increase in blood pressure nor any increase in [3H]MeTRH binding in the hypothalamus and striatum as compared to age matched WKY rats. With the development of elevated blood pressure at 6 weeks, an increase in [3H]MeTRH binding in the hypothalamus and striatum of SHR rats in comparison to the tissues from WKY rats was observed. The results provide, for the first time, evidence for a parallel increase in the density of brain TRH receptors with elevation of blood pressure, and suggest that brain TRH receptors may play an important role in the pathophysiology of hypertension. PMID- 3035514 TI - Selective activation of opioid receptors differentially affects lordosis behavior in female rats. AB - The effects of opioid peptides that are highly selective ligands for mu receptors (morphiceptin). delta receptors (delta-receptor peptide), kappa receptors (dynorphin 1-9), and the mu/delta complex (beta-endorphin), were tested on lordosis behavior in ovariectomized rats primed with estrogen and progesterone. Intracerebroventricular infusions of beta-endorphin or morphiceptin both inhibited and facilitated lordosis in a dose-dependent fashion whereas all doses of delta-receptor peptide facilitated lordosis. Dynorphin 1-9 had no significant effect at any dose, although a trend toward increased lordosis quotients was observed 30 min after infusion. The effects of beta-endorphin, morphiceptin, and delta-receptor peptide were reversed with naloxone, although naloxone alone had no effect on lordosis behavior. These results indicate that the specific activation of opioid receptor subtypes differentially affects lordosis behavior. It appears that binding to high-affinity mu 1 receptors exerts an inhibitory influence on lordosis, whereas binding to low-affinity mu 2 receptors or delta receptors exerts a facilitatory influence. Binding to kappa receptors does not appear to affect lordosis behavior. PMID- 3035515 TI - Modulation of dopamine receptors by thyrotropin-releasing hormone in the rat brain. AB - Nanomolar concentration of thyrotropin-releasing hormone (TRH) in vitro caused a significant reduction of [3H]apomorphine binding sites (70% of the control) in the rat striatum and the limbic forebrain. [3H]Spiperone binding was not affected by TRH. On the other hand, dopamine and apomorphine displaced [3H]TRH binding partially, suggesting the presence of a TRH receptor subpopulation that has a high affinity for dopamine agonist. Most of the neuroleptics displaced [3H]TRH binding dose-dependently in the micromolar range. (-)-Sulpiride had no affinity to TRH receptors. These findings suggest that one of the important roles of TRH as a neuromodulator is to modulate receptors for classical neurotransmitters, and this receptor-receptor interaction may be of importance in explaining the well known stimulating effects of TRH on the dopaminergic system. PMID- 3035518 TI - [Mechanisms of action and clinical use of converting enzyme inhibitors]. PMID- 3035520 TI - [Acquired immunodeficiency syndrome: clinical picture and changes in various organs (illustrated by description of a case)]. PMID- 3035517 TI - [Role of tin (II) and pyrophosphate in the process of technetium-99m binding in erythrocytes]. PMID- 3035519 TI - [Enalapril in the treatment of primary hypertension: its effect on blood pressure and plasma renin activity]. PMID- 3035521 TI - Primary signet-ring cell carcinoma of the urinary bladder. AB - An unusual case of the signet-ring cell carcinoma of the bladder was reported. A 62-year-old Japanese male had a three-month history of intermittent, painless and gross hematuria. He was found to have a small egg-sized bladder tumor near the right ureteral orifice. A cytologic diagnosis of signet-ring cell carcinoma was made on a bladder washing and was confirmed by cytoscopic biopsy and surgical specimens. PMID- 3035516 TI - [Binding of tin (II) and technetium-99m in subcellular fractions of erythrocytes]. PMID- 3035522 TI - Immunohistochemical demonstration of lacto-N-fucopentose III in lung carcinomas with monoclonal antibody 624A12. AB - Bronchopulmonary carcinomas were analyzed immunohistochemically using monoclonal antibody 624A12. The antibody was raised against a human "small cell carcinoma" cell line NCI-H69. It recognizes a particular sugar sequence in lacto-N fucopentose III, which is preserved in formalin fixed and paraffin embedded tissue. Various bronchopulmonary carcinomas revealed characteristic patterns of immunoreactivity. Forty nine/50 adenocarcinomas were immunoreactive either diffusely or focally. The immunostaining was usually limited to the cell membranes with occasional intracytoplasmic immunostaining in large cells. The only negative case had been irradiated before surgical resection. Twenty seven/38 squamous cells carcinomas did not immunostain while the remaining 11 displayed focal immunoreactivity in areas of "loose cellular apposition" associated with necrosis and, rarely, in squamous pearls. All of six adenosquamous carcinomas showed immunoreactivity focally. Eleven/30 large cell carcinomas and 10/11 bronchiolo-alveolar carcinomas were either diffusely or focally immunoreactive. Seven/26 intermediate cell neuroendocrine carcinomas were focally immunoreactive while none of 33 typical small cell neuroendocrine carcinomas, 21 carcinoids, and 10 well differentiated neuroendocrine carcinomas was immunoreactive. An adenoid cystic carcinoma was diffusely immunoreactive, and a mucoepidermoid carcinoma was focally immunoreactive. We conclude that various bronchopulmonary neoplasms have characteristic patterns of distribution of this antigen, and that monoclonal antibody 624A12 may be useful for the differential diagnose among bronchopulmonary carcinomas, and their differential diagnosis from pleural mesotheliomas. PMID- 3035523 TI - Neonatal cytomegalovirus infection with pancreatic cystadenoma and nephrotic syndrome. AB - An infant with microcephaly and generalized cytomegalovirus infection presented with two unusual complications: cystadenoma of the pancreas and minimal change nephrotic syndrome (MCN). Possible pathogenetic mechanisms of these conditions are discussed. PMID- 3035524 TI - Congenital fibrosarcoma: a congenital fibrous histiocytoma. AB - Using immunoperoxidase PAP technique in 2 cases of congenital fibrosarcoma, a great number of cells showed positive stain for alpha-1-antitrypsin (A1AT) and alpha-1-antichymotrypsin (A1ACT), both considered to be good histiocytic markers. The ultrastructure in 1 case also provides evidence of histiocytic differentiation. These findings suggest that congenital fibrosarcomas are actually congenital fibrous histiocytomas and may explain the presence of inflammatory infiltrates as in some malignant fibrous histiocytomas. PMID- 3035525 TI - Treatment of congestive heart failure. Pathophysiologic and pharmacologic concepts. AB - Congestive heart failure is a serious health problem in Western society. Understanding of both the basic pathophysiology and the design of rational and effective treatment of this syndrome has increased dramatically in the last ten years. However, congestive heart failure remains a highly lethal disease, which kills at a rate exceeding that of most cancers. Prevention is therefore of the utmost importance and has to come primarily from a reduction in the incidence and damage done from myocardial infarction. Improved understanding of the pathophysiology of idiopathic dilated cardiomyopathy is also needed. Through understanding of pathophysiologic principles, effective therapy based on interference with known mechanisms that are operative in the disease is now available but can be refined in the future. PMID- 3035526 TI - Gene insertion into the chicken germ line by retroviruses. AB - We injected chick syncytial strain of reticuloendotheliosis virus (CS-REV) and wild type and recombinant avian leukosis virus (ALV) near the blastoderm of unincubated fertilized embryos and CS-REV intra-abdominally at day of hatch, and we progeny tested the surviving ALV viremic males and REV viremic males and females for transmitted viral genetic material. A number of positive progeny were identified and their deoxyribonucleic acid (DNA) analyzed for restriction enzyme fragments that hybridized with viral genetic material. Most of the progeny had simple restriction enzyme patterns unlike the viremic parents or congenitally infected progeny. This is suggestive evidence that retroviral genetic information has been inserted into the germ line of chickens. PMID- 3035527 TI - Design of retroviral vectors for the insertion of foreign deoxyribonucleic acid sequences into the avian germ line. AB - Because the available avian leukosis viral (ALV) vectors are moderately oncogenic in vivo, they are not suitable for insertion into the germ line. A significant reduction in the oncogenicity of the ALV vectors can be achieved by substituting the noncoding long terminal repeats (LTR) regions of the ALV virus with the LTR of the nononcogenic endogenous RAV-O virus. There is good evidence that the resulting RAV-O LTR vectors can be inserted into the germ line of domestic chickens and have the potential for inserting cloned sequences that can be used for poultry improvement. PMID- 3035528 TI - Effects of feeding graded levels of vitamin D3 on egg shell pimpling in aged hens. AB - Pimpled egg shells are one of the various types of egg shell problems in the industry today. In this study, graded levels of vitamin D3 (D3) (0, 138, 275, 550, 2200, 22,000, 44,000, and 88,000 ICU of D3/kg of feed) were fed in a corn soy ration to Single Comb White Leghorn hens (78 weeks of age), randomly assigned to one of the eight treatments. Feed consumption, egg production, egg weight, egg specific gravity, and pimple score were determined at weekly or biweekly intervals for a 10-week period. At termination of the experiment, calcium, sodium, potassium, and ash content of the uterus and serum calcium levels were assayed. Results showed that a significant (P less than .05) linear increase in pimple score, uterine ash, and serum calcium occurred as dietary levels of D3 increased. Egg production and egg specific gravity were correlated to D3 level in a quadratic fashion. Uterine sodium levels declined with increased D3 in the diet, while potassium values showed no differences. Calcium levels of uterine tissue tended to increase with increased D3. The lowest (0 ICU/kg) and highest level of D3 (88,000 ICU/kg) significantly decreased feed consumption. It was concluded that egg shell pimpling is directly related to level of cholecalciferol (D3) in the diet. PMID- 3035529 TI - The effect of mercury chloride and methyl mercury on brain microsomal Na+-K+ ATPase after partial delipidisation with Lubrol. AB - The microsomal Na+-K+-ATPase of rat brain was inhibited by mercury chloride and methyl mercury. The IC50 was 6.5 X 10(-7) M for mercury chloride and 3.5 X 10(-6) M for methyl mercury. The inhibition was of a non-competitive type with respect to ATP. The non-ionic detergent Lubrol potentiated the inhibitory effect of both mercurials. It is concluded that Lubrol removes the bulk lipids present outside the catalytic center of the enzyme. Consequently, the enzyme will become more sensitive to the inhibition by both mercurials. PMID- 3035530 TI - Non-adrenergic, non-cholinergic parasympathetic secretion in the rat submaxillary and sublingual glands. AB - Atropine resistant secretion from the submaxillary and sublingual glands was demonstrated upon electrical stimulation of the chorda-lingual nerve at high frequencies. The flow rate of the protein-rich saliva was low, and it declined rapidly and markedly upon prolonged stimulation. The results show that one or more substances other than acetylcholine are released by parasympathetic stimulation and that they may be of importance for the regulation of the exocrine functions of the glands. When the continuous mode of electrical stimulation at a low frequency of the nerve was changed to an intermittent mode of stimulation at a high frequency (in the absence of atropine), the secretion of fluid and protein decreased. PMID- 3035531 TI - Chronic salt loading and adrenergic mechanisms in the Sprague-Dawley rat. AB - The effect of chronic salt loading on adrenergic mechanisms was evaluated in Sprague-Dawley rats (and NMRI mice) maintained on a high sodium (8%) or normal sodium (0.3%) regime for 4 weeks. The basal blood pressure (carotid artery) was not influenced by the high salt diet but the heart rate and blood pressure increases to mental stress (jet air) were larger in the salt loaded rats. There were indications of an increased sympathetic tone in rats on the high salt diet since in these rats sympathoinhibitory treatment with ganglionic blockade or clonidine induced larger falls in blood pressure and heart rate than in the controls. Central catecholamines (brain stem, striatum, hemispheres) were determined spectrofluorimetrically after cation exchange chromatography. The high salt diet influenced neither the endogenous levels of noradrenaline nor central noradrenaline turnover (disappearance of noradrenaline after synthesis inhibition by alpha-methyltyrosine and accumulation of dihydroxyphenylalanine after decarboxylase inhibition by 3-hydroxybenzylhydrazine). There were no changes in central alpha 2-adrenoceptor responsiveness when assessed as clonidine-induced deceleration of noradrenaline turnover in the brain and in central alpha 1 adrenoceptor responsiveness (clonidine-induced increase of flexor reflex in spinalized rats and clonidine-induced increase of motor activity in reserpinized mice). Peripheral sympathetic function was assessed in pithed rats. The pressor responses to intravenously administered noradrenaline (0.01-10 micrograms/kg) and electrical stimulation of the spinal sympathetic nerves (SNS, 0.25-2 Hz) were similar in the two groups, suggesting that salt did not influence vascular alpha adrenoceptor responsiveness or transmitter release.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3035533 TI - Extracellular phytase (E.C. 3.1.3.8) from Aspergillus ficuum NRRL 3135: purification and characterization. AB - Extracellular phytase from Aspergillus ficuum, a glycoprotein, was purified to homogeneity in 3 column chromatographic steps using ion exchange and chromatofocusing. Results of gel filtration chromatography and SDS-polyacrylamide gel electrophoresis indicated the approximate molecular weight of the native protein to be 85-100-KDa. On the basis of a molecular weight of 85-KDa, the molar extinction coefficient of the enzyme at 280 nm was estimated to be 1.2 X 10(4) M 1 cm-1. The isoelectric point of the enzyme, as deduced by chromatofocusing, was about 4.5. The purified enzyme is remarkably stable at 0 degree C. Thermal inactivation studies have shown that the enzyme retained 40% of its activity after being subjected to 68 degrees C for 10 minutes, and the enzyme exhibited a broad temperature optimum with maximum catalytic activity at 58 degrees C. The Km of the enzyme for phytate and p-nitrophenylphosphate is about 40 uM and 265 uM, respectively, with an estimated turnover number of the enzyme for phytate of 220 per sec. Enzymatic deglycosylation of phytase by Endoglycosidase H lowered the molecular weight of native enzyme from 85-100-KDa to about 76-KDa; the digested phytase still retained some carbohydrate as judged by positive periodic acid Schiff reagent staining of the electrophoresed protein. Immunoblotting of the phytase with monoclonal antibody 7H10 raised against purified native enzyme recognized not only native but also partially deglycosylated protein. PMID- 3035532 TI - Prenatal diagnosis of Fabry's disease by direct analysis of chorionic villi. AB - Six pregnancies of three carriers for X-linked Fabry's disease, were monitored by chromosome and enzyme analysis. Two affected male fetuses were detected by the demonstration of alpha-galactosidase deficiency in amniotic fluid cells and chorionic villi respectively. The use of chorionic villi enabled a diagnosis within a few hours after sampling in the ninth week of pregnancy whereas the use of amniotic fluid cells in the earlier case required two weeks of culturing after amniocentesis in the 16th week. Four female fetuses were found; heterozygosity was demonstrated in one by analysis of clones in the primary amniotic fluid cell culture. PMID- 3035534 TI - [A secondary effect of converting enzyme inhibitors: cough]. AB - Cough associated with angiotensin-converting enzyme (ACE) inhibitors has long been considered a rare side effect. We report 8 cases, 7 with enalapril (10 to 20 mg/day) 1 with quinapril (40 mg/day) in which cough occurred after a mean duration of treatment of 39 days. In all patients, cough disappeared with a mean delay of 2 days with no other treatment than withdrawal of the drug. In 6 patients, cough was reinduced within less than a day with the same drug; in 5 patients a second reinduction with another ACE inhibitor gave the same result. These data suggest that cough is probably more frequent than it would appear from the literature. In clinical practice, if cough occurs in a patient treated with an ACE inhibitor, the drug may be continued for a few days in order to exclude an acute viral infection; if cough lasts more than a week, specific diagnostic procedures for pulmonary disease should be initiated; if it stops, the patient may be treated either for hypertension or chronic heart failure with another ACE inhibitor. PMID- 3035535 TI - Detection of Leishmania parasites by DNA in situ hybridization with non radioactive probes. AB - In situ hybridization techniques develop rapidly into diagnostic tools of considerable value for detection of viruses and bacteria. Here we report the application of this technique for the detection of Leishmania parasites. Biotin labelled total promastigote DNA was hybridized to cultured Leishmania parasites and to blood and impression smears of infected mice. In promastigotes kinetoplasts were strongly stained, nuclei somewhat more diffuse. In amastigotes both nuclear and kinetoplast DNA hybridized strongly. Amastigotes were easily detected in tissue of infected mice by their stable configuration of kinetoplast and nuclei. Cross-hybridization was observed between Leishmania donovani and L. tropica, but not between these two and L. braziliensis or Trypanosoma cruzi. A minor aspecific staining of host cell nuclei in the smears did not interfere with the detectability of the parasites. PMID- 3035536 TI - Isolation and characterization of full-length cDNA clones coding for cholinesterase from fetal human tissues. AB - To study the primary structure and regulation of human cholinesterases, oligodeoxynucleotide probes were prepared according to a consensus peptide sequence present in the active site of both human serum pseudocholinesterase (BtChoEase; EC 3.1.1.8) and Torpedo electric organ "true" acetylcholinesterase (AcChoEase; EC 3.1.1.7). Using these probes, we isolated several cDNA clones from lambda gt10 libraries of fetal brain and liver origins. These include 2.4 kilobase cDNA clones that code for a polypeptide containing a putative signal peptide and the N-terminal, active site, and C-terminal peptides of human BtChoEase, suggesting that they code either for BtChoEase itself or for a very similar but distinct fetal form of cholinesterase. In RNA blots of poly(A)+ RNA from the cholinesterase-producing fetal brain and liver, these cDNAs hybridized with a single 2.5-kilobase band. Blot hybridization to human genomic DNA revealed that these fetal BtChoEase cDNA clones hybridize with DNA fragments of the total length of 17.5 kilobases, and signal intensities indicated that these sequences are not present in many copies. Both the cDNA-encoded protein and its nucleotide sequence display striking homology to parallel sequences published for Torpedo AcChoEase. These findings demonstrate extensive homologies between the fetal BtChoEase encoded by these clones and other cholinesterases of various forms and species. PMID- 3035537 TI - Nucleoside triphosphate-dependent DNA-binding properties of mos protein. AB - We have previously shown that the mos gene product, p40mos, produced in Escherichia coli binds ATP and has ATPase activity. In the present study, we investigated the DNA-binding properties of p40mos and two mos deletion mutant proteins. Nitrocellulose blot protein-DNA binding assays showed that p40mos binds DNA in the presence of Mg2+-ATP and certain other nucleoside triphosphates. Ninety percent of the p40mos-bound DNA is dissociated if the complex is washed in the presence of 1 M NaCl or in the absence of ATP. p40mos-DNA binding is not observed in the presence of AMP or the nonhydrolyzable ATP analog adenosine 5' [beta, gamma-methylene]-triphosphate; however, in the presence of ADP, p40mos binds DNA at 20% of the level that is observed with ATP. An N-terminal-deletion mutant protein, p19mos, has no DNA-binding activity, whereas a C-terminal deletion mutant protein, p25mos, does. p25mos contains the ATP-binding domain, binds DNA in the presence of either ADP or ATP, and shows 5% and 45% binding (relative to that in the presence of ATP) in the presence of AMP and adenosine 5' [beta, gamma-methylene]triphosphate, respectively. These results suggest that the N-terminal domain of p40mos is responsible for nucleoside triphosphate-mediated DNA binding. We also observed differential histone-DNA binding in the presence and absence of ATP. PMID- 3035538 TI - Analysis and in vivo disruption of the gene coding for calmodulin in Schizosaccharomyces pombe. AB - Calmodulin is a low molecular weight calcium-binding protein that modulates many enzyme systems in eukaryotes. We have cloned the gene encoding calmodulin from the fission yeast, Schizosaccharomyces pombe, by using synthetic oligonucleotide probes that correspond to three distinct regions of Tetrahymena calmodulin. A 1.6 kilobase (kb) DNA fragment that hybridized to all of them contains a gene whose deduced product possesses 74% amino acid homology with bovine calmodulin. This gene, which is unique in the S. pombe genome and is named cam1, encodes 149 amino acids excluding the first methionine and is transcribed into mRNA of 1.2-kb length. It has an intron that apparently starts immediately after the initiation codon and is 126 bp long. S. pombe calmodulin exhibits more homology to vertebrate calmodulin than to that of the budding yeast, Saccharomyces cerevisiae. Gene disruption experiments revealed that cam1 gene function is essential for vegetative growth of S. pombe. Spores bearing disrupted cam1 halt growth soon after germination and rarely carry out the first cell division, indicating that calmodulin does not exist in excess in those cells. PMID- 3035539 TI - Endogenous inhibitor for calcium-dependent cysteine protease contains four internal repeats that could be responsible for its multiple reactive sites. AB - A cDNA encoding an endogenous inhibitor, termed calpastatin, for calcium dependent cysteine protease (calpain, EC 3.4.22.17) was cloned by screening rabbit cDNA libraries with a synthetic oligodeoxynucleotide probe based on the partial amino acid sequence of the purified protein. The deduced amino acid sequence contains 718 amino acid residues (Mr, 76,964), and the mature protein corresponds to the deduced sequence from the 80th residue of the primary translation product (resultant Mr, 68,113). This deduced molecular weight is significantly lower than that determined by NaDodSO4/polyacrylamide gel electrophoresis, suggesting the possibility that the inhibitor is post translationally modified. The sequence of the mature inhibitor contains four consecutive internal repeats approximately 140 amino acid residues long, each of which might be responsible for the inhibitory activity. Calpastatin is apparently different from a typical cysteine protease inhibitor (cystatin), suggesting that the mechanism of inhibition of calcium-dependent cysteine protease by the inhibitor might be different from that of other cysteine proteases by cystatin. PMID- 3035541 TI - Molecular cloning of cDNAs of human liver and placenta NADH-cytochrome b5 reductase. AB - A cDNA coding for human liver NADH-cytochrome b5 reductase (cytochrome b5 reductase, EC 1.6.2.2) was cloned from a human liver cDNA library constructed in phage lambda gt11. The library was screened by using an affinity-purified rabbit antibody against NADH-cytochrome b5 reductase of human erythrocytes. A cDNA about 1.3 kilobase pairs long was isolated. By using the cDNA as a probe, another cDNA (pb5R141) of 1817 base pairs was isolated that hybridized with a synthetic oligonucleotide encoding Pro-Asp-Ile-Lys-Tyr-Pro, derived from the amino acid sequence at the amino-terminal region of the enzyme from human erythrocytes. Furthermore, by using the pb5R141 as a probe, cDNA clones having more 5' sequence were isolated from a human placenta cDNA library. The amino acid sequences deduced from the nucleotide sequences of these cDNA clones overlapped each other and consisted of a sequence that completely coincides with that of human erythrocytes and a sequence of 19 amino acid residues extended at the amino terminal side. The latter sequence closely resembles that of the membrane-binding domain of steer liver microsomal enzyme. PMID- 3035542 TI - Helicase properties of the Escherichia coli UvrAB protein complex. AB - The Escherichia coli UvrA protein has an associated ATPase activity with a turnover number affected by the presence of UvrB protein as well as by DNA. Specifically, the structure of DNA significantly influences the turnover rate of the UvrAB ATPase activity. Double-stranded DNA maximally activates the turnover rate 10-fold whereas single-stranded DNA maximally activates the turnover rate 20 fold, suggesting that the mode of interaction of UvrAB protein with different DNAs is distinctive. We have previously shown that the UvrAB protein complex, driven by the binding energy of ATP, can locally unwind supercoiled DNA. The nature of the DNA unwinding activity and single-stranded DNA activation of ATPase activity suggests potential helicase activity. In the presence of a number of helicase substrates, the UvrAB complex, indeed, manifests a strand-displacement activity--unwinding short duplexes and D-loop DNA, thereby generating component DNA structures. The energy for the activity is derived from ATP or dATP hydrolysis. Unlike the E. coli DnaB, the UvrAB helicase is sensitive to UV induced photoproducts. PMID- 3035540 TI - Characteristics of binding of human seminal alpha-inhibin-92 to human pituitary membranes. AB - We investigated the binding of 125I-labeled alpha-inhibin-92 (a 92-residue peptide) to human pituitary membrane preparations. Unlabeled alpha-inhibin-92 competed effectively with the labeled peptide for binding to the membranes. Binding was also inhibited by both alpha-inhibin-52 and alpha-inhibin-31, but less effectively. Scatchard analysis of the alpha-inhibin-92 binding data indicated the presence of high-affinity binding sites (1.35 nM/mg of membrane protein) with an apparent Kd of 0.37 nM. When 125I-labeled alpha-inhibin-92 was covalently crosslinked to the pituitary membrane preparation with disuccinimidyl suberate and the solubilized labeled receptor complex was analyzed by NaDodSO4/PAGE under either reducing or nonreducing conditions, a single radioactive band at an apparent molecular weight of 90,000 +/- 5000 was observed. These data suggest that human pituitary has specific binding sites for alpha inhibins. PMID- 3035543 TI - Initiation of simian virus 40 DNA replication in vitro: large-tumor-antigen- and origin-dependent unwinding of the template. AB - Analysis of the kinetics of simian virus 40 (SV40) DNA replication in vitro demonstrated the existence of a slow presynthesis reaction that occurs prior to onset of extensive chain elongation and is dependent on a subset of the cellular proteins required for the complete replication reaction. When the presynthesis reaction is carried out in the presence of topoisomerase I, it is possible to detect extensive unwinding of the template DNA. This unwinding reaction is specific for templates that contain the wild-type SV40 origin of DNA replication and requires SV40 large tumor antigen (T antigen), ATP, and a protein fraction derived from HeLa cells. The required cellular protein may be a eukaryotic single stranded-DNA-binding protein (SSB), since unwinding of the template is also observed when Escherichia coli SSB is substituted for the HeLa protein fraction. These observations suggest that during the initial stages of SV40 DNA replication, T antigen binds specifically to the viral origin and locally unwinds the DNA. This origin-dependent unwinding reaction is presumably a prerequisite for subsequent priming and elongation steps. PMID- 3035544 TI - Transcriptional (p40x) and post-transcriptional (p27x-III) regulators are required for the expression and replication of human T-cell leukemia virus type I genes. AB - The pX sequence of human T-cell leukemia virus type I codes for three products: p40x, p27x-III, and p21x-III. p40x is a transcriptional trans-activator that activates not only the viral long terminal repeat but also cellular genes for interleukin 2 and its receptor. p27x-III and p21x-III are not required for transcriptional activation, and their functions were unknown. Cotransfection experiments with defective human T-cell leukemia virus type I proviruses and various pX expression plasmids revealed that p27x-III, in addition to p40x, was required for gag gene expression. Furthermore, it was shown that p27x-III induced accumulation of a high level of unspliced viral gag mRNA. These results indicate that p27x-III is a post-transcriptional modulator of viral RNA whose transcription has been fully activated by p40x. PMID- 3035545 TI - Binding of transcription factors and creation of a large nucleoprotein complex on the human cytomegalovirus enhancer. AB - The effect of the human cytomegalovirus immediate early region 1 enhancer on transcription was studied in vitro with HeLa cell nuclear extract. Stimulation of in vitro transcription mediated by the enhancer element involves its recognition by specific trans-acting factors present in the nuclear extract. DNase I protection analysis was used to determine at the nucleotide level those enhancer sequences that interact with nuclear factors. At least nine sites of protein-DNA interaction were detected over approximately 400 base pairs of enhancer sequence. The regions of nuclease protection are associated with 21-, 19-, 18-, and 17-base pair repeat elements as well as with a unique sequence, creating a large nucleoprotein complex. The relationship between the protein binding and the activity of the immediate early region 1 enhancer is discussed. PMID- 3035547 TI - Ubiquitin has intrinsic proteolytic activity: implications for cellular regulation. AB - Ubiquitin is a protein of 76 amino acids found in every eukaryotic cell. Although ubiquitin is implicated in ATP-dependent nonlysosomal protein degradation and is also conjugated to specific cellular proteins, the role played by ubiquitin in cellular events has not been defined. We report that purified ubiquitin has intrinsic proteolytic activity and demonstrate that this activity is comparable to that of other well-characterized proteases. Monoclonal antibodies specific to ubiquitin inhibit proteolysis. Ubiquitin has protease activity over a broad pH range with an optimum at pH 8.0. It is stimulated by Ca2+ and is inhibited by high concentrations of phenylmethylsulfonyl fluoride and diisopropyl fluorophosphate. Ubiquitin will cleave proteins at a limited number of sites. We propose that the ubiquitination of a protein can convert that protein into an ad hoc specific protease and models are presented as to how this can play a role in regulating a variety of cellular events. PMID- 3035546 TI - Replication of mini-P1 plasmid DNA in vitro requires two initiation proteins, encoded by the repA gene of phage P1 and the dnaA gene of Escherichia coli. AB - We have developed an in vitro DNA-replication system that replicates exogenously added mini-P1 plasmid DNA. The system consists of purified P1 RepA protein and a partially purified mixture of Escherichia coli replication proteins. It is essentially the same as that described for the replication of oriC plasmid DNA [Fuller, R.S., Kaguni, J.M. & Kornberg, A. (1981) Proc. Natl. Acad. Sci. USA 78, 7370-7374]. Mini-P1 DNA replication requires the E. coli DnaA initiation protein in addition to the P1 RepA initiation protein. The reaction is inhibited by rifampicin, novobiocin, and antibody to DnaB, suggesting the involvement of RNA polymerase, DNA gyrase, and DnaB protein. Replication is initiated in the region of the P1 origin of replication and proceeds unidirectionally as determined by electron microscopy. Thus, the in vitro system mimics the essential features of mini-P1 replication as suggested by genetic studies. PMID- 3035549 TI - Regulation of the host range of human papovavirus JCV. AB - Human papovavirus JCV is associated with the human demyelinating disorder progressive multifocal leukoencephalopathy. In tissue culture, the virus is largely restricted to growth in primary human fetal glial cell. In this study, we demonstrate two levels of regulation of the viral host range. Expression of the early JCV mRNA, which encodes the essential viral protein, large tumor antigen (T antigen), depends on recognition of the early enhancer/promoter elements by tissue-specific factors found in both human and rodent glial cells. In the presence of JCV T antigen, viral DNA replication requires a species-specific factor, presumably a component of DNA polymerase, which is found in a wide range of primate cells. We further demonstrate that simian virus 40 T antigen has sufficient homology to efficiently substitute for the analogous JCV protein in initiating viral DNA replication. PMID- 3035548 TI - Identification of the receptor for erythropoietin by cross-linking to Friend virus-infected erythroid cells. AB - Erythropoietin (Epo) is a glycoprotein hormone that regulates erythroid development and interacts with surface receptors on developing erythroid cells. In this laboratory, a cell system with a relatively pure population of erythroid cells that respond to Epo has been developed. Immature erythroid cells are obtained from the spleens of mice infected with the anemia strain of Friend virus. The binding of 125I-labeled Epo (125I-Epo) to plasma membranes from these cells was studied in this investigation. 125I-Epo binding reached equilibrium within 20 min at 37 degrees C. Twenty percent of the receptors bound 125I-Epo with a Kd of 0.08 X 10(-9) M, while the remaining receptors bound the hormone with a Kd of 0.6 X 10(-9) M. In this study, a receptor for Epo was identified by cross-linking 125I-Epo to the receptor in intact cells and plasma membrane preparations using disuccinimidyl suberate. Polyacrylamide gel electrophoresis revealed two labeled bands of 100 and 85 kDa. The 85-kDa band was more heavily labeled (65%) than the 100-kDa band. Both bands were equally decreased when increasing amounts of unlabeled Epo were included in the binding mixture, indicating a specific interaction of 125I-Epo with the receptor. PMID- 3035550 TI - Prostaglandin F2 alpha stimulates phosphatidylinositol 4,5-bisphosphate hydrolysis and mobilizes intracellular Ca2+ in bovine luteal cells. AB - The present studies were conducted to determine whether prostaglandin F2 alpha (PGF2 alpha) stimulates the production of "second messengers" derived from inositol phospholipid hydrolysis and increases intracellular free Ca2+ ([Ca2+]i) in isolated bovine luteal cells. PGF2 alpha provoked rapid (10 sec) and sustained (up to 60 min) increases in the levels of inositol mono-, bis-, and trisphosphates (InsP, InsP2, and InsP3, respectively). InsP3 was formed more rapidly than InsP2 or InsP after PGF2 alpha treatment. In addition, PGF2 alpha increased inositol phospholipid turnover, as evidenced by increased 32PO4 incorporation into phosphatidic acid and phosphatidylinositol. LiCl (1-20 mM) enhanced inositol phosphate accumulation in response to PGF2 alpha. Maximal increases in InsP3 occurred at 1 microM PGF2 alpha, with half-maximal stimulation occurring at 36 nM. The acute effects of PGF2 alpha on InsP3 levels were independent of reductions in extracellular calcium. Prostaglandins E1 and E2 also stimulated increases in inositol phosphate levels, albeit to a lesser extent. PGF2 alpha also induced rapid and concentration-dependent increases in [Ca2+]i as measured by quin-2 fluorescence. The PGF2 alpha-induced increases in [Ca2+]i were maximal within 30 sec (approximately 2- to 3-fold), and [Ca2+]i remained elevated for 8-10 min. The PGF2 alpha-induced increases in [Ca2+]i were also independent of extracellular calcium. These findings demonstrate that the action of PGF2 alpha is coupled to the phospholipase C-InsP3 and diacylglycerol second messenger system in the corpus luteum. PMID- 3035551 TI - The human transforming growth factor type alpha coding sequence is not a direct acting oncogene when overexpressed in NIH 3T3 cells. AB - A peptide secreted by some tumor cells in vitro imparts anchorage-independent growth to normal rat kidney (NRK) cells and has been termed transforming growth factor type alpha (TGF-alpha). To directly investigate the transforming properties of this factor, the human sequence coding for TGF-alpha was placed under the control of either a metallothionein promoter or a retroviral long terminal repeat. These constructs failed to induce morphological transformation upon transfection of NIH 3T3 cells, whereas viral oncogenes encoding a truncated form of its cognate receptor, the EGF receptor, or another growth factor, sis/platelet-derived growth factor 2, efficiently induced transformed foci. When NIH 3T3 clonal sublines were selected by transfection of TGF-alpha expression vectors in the presence of a dominant selectable marker, they were shown to secrete large amounts of TGF-alpha into the medium, to have downregulated EGF receptors, and to be inhibited in growth by TGF-alpha monoclonal antibody. These results indicated that secreted TGF-alpha interacts with its receptor at a cell surface location. Single cell-derived TGF-alpha-expressing sublines grew to high saturation density in culture. However, when plated as single cells on contact inhibited monolayers of NIH 3T3 cells, they failed to form colonies, whereas v sis- and v-erbB-transfected cells formed transformed colonies under the same conditions. Moreover, TGF-alpha-expressing sublines were not tumorigenic in nude mice. These and other results imply that TGF-alpha exerts a growth-promoting effect on the entire NIH 3T3 cell population after secretion into the medium but little, if any, effect on the individual cell synthesizing this factor. It is concluded that the normal coding sequence for TGF-alpha is not a direct-acting oncogene when overexpressed in NIH 3T3 cells. PMID- 3035552 TI - Nonpolarized secretion of truncated forms of the influenza hemagglutinin and the vesicular stomatitus virus G protein from MDCK cells. AB - The demonstration that the envelope glycoproteins G of vesicular stomatitus virus and hemagglutinin of influenza virus synthesized in polarized epithelial cells transfected with the corresponding genes are effectively segregated to the basolateral or apical plasma membrane domains, respectively, implies that the information determining this segregation resides within the structures of the proteins themselves. To localize the sorting information within these proteins, the polarity of secretion of truncated hemagglutinin and G glycoproteins secreted from confluent monolayers of MDCK cells transformed with vectors containing the corresponding truncated cDNAs was examined. It was found that, even though the transformed cells continued to secrete a major endogenous glycoprotein exclusively from the apical surface, the modified viral glycoproteins were secreted in a nonpolarized fashion from both sides of the monolayers. These observations suggest that important information for the sorting of the viral glycoprotein is contained within their membrane anchoring or cytoplasmic segments or that, if sorting signals are luminally located, these signals must be present in a conformation that is not attainable when the polypeptides are not attached to the membrane. PMID- 3035553 TI - Nucleotide sequence and evolution of ETn elements. AB - The ETn (for "early transposon") family of long repeated sequences in abundantly transcribed in early mouse embryos from retroviral-like long terminal repeats. Nucleotide sequencing of two elements does not reveal any long open reading frame nor significant homology to retroviral proteins. The genetic polymorphism, monitored by Southern blotting within and across mouse species, reflects a concerted mode of evolution for the ETn sequences. PMID- 3035554 TI - Characterization of five partial deletions of the factor VIII gene. AB - Hemophilia A is an X-linked disorder of coagulation caused by a deficiency of factor VIII. By using cloned DNA probes, we have characterized the following five different partial deletions of the factor VIII gene from a panel of 83 patients with hemophilia A: (i) a 7-kilobase (kb) deletion that eliminates exon 6; (ii) a 2.5-kb deletion that eliminates 5' sequences of exon 14; (iii) a deletion of at least 7 kb that eliminates exons 24 and 25; (iv) a deletion of at least 16 kb that eliminates exons 23-25; and (v) a 5.5-kb deletion that eliminates exon 22. The first four deletions are associated with severe hemophilia A. By contrast, the last deletion is associated with moderate disease, possibly because of in frame splicing from moderate disease, possibly because of in-frame splicing from adjacent exons. None of those patients with partial gene deletions had circulating inhibitors to factor VIII. One deletion occurred de novo in a germ cell of the maternal grandmother, while a second deletion occurred in a germ cell of the maternal grandfather. These observations demonstrate that de novo deletions of X-linked genes can occur in either male or female gametes. PMID- 3035555 TI - Genomic organization of the mouse T-cell receptor beta-chain gene family. AB - We have combined three different methods, deletion mapping of T-cell lines, field inversion gel electrophoresis, and the restriction mapping of a cosmid clone, to construct a physical map of the murine T-cell receptor beta-chain gene family. We have mapped 19 variable (V beta) gene segments and the two clusters of diversity (D beta) and joining (J beta) gene segments and constant (C beta) genes. These members of the beta-chain gene family span approximately equal to 450 kilobases of DNA, excluding one potential gap in the DNA fragment alignments. PMID- 3035556 TI - Amyloid fibrils in human insulinoma and islets of Langerhans of the diabetic cat are derived from a neuropeptide-like protein also present in normal islet cells. AB - Amyloid deposits localized to the islets of Langerhans are typical of non-insulin dependent human diabetes mellitus and of diabetes mellitus in adult cats. Amyloid deposits also commonly occur in insulin-producing pancreatic tumors. We have purified a major protein--insulinoma or islet amyloid polypeptide (IAPP)--from human and cat islet amyloid and from amyloid of a human insulinoma. IAPP from human insulinoma contained 37 amino acid residues and had a theoretical molecular mass of 3850 Da. The amino acid sequence is unique but has greater than 40% identity with the human calcitonin gene-related peptide. A partial amino acid sequence of cat islet IAPP corresponding to positions 1-27 of human insulinoma IAPP was identical to the human IAPP except for substitutions in three positions. An antiserum raised to a synthetic human insulinoma IAPP-(7-17) undecapeptide showed specific immunohistochemical reactivity with human and cat islet amyloid and with islet B cells. The significance of this pancreatic neuropeptide-like protein is unknown, but it is suggested that it may exert an important endocrine regulatory effect. PMID- 3035557 TI - Varicella-zoster virus as a live vector for the expression of foreign genes. AB - The previous demonstration of the efficacy and tolerability of the Oka strain of varicella-zoster virus (VZV) in clinical trials involving vaccination of both normal and immunocompromised individuals has laid the foundation for its use in preventing chickenpox. In this context, VZV could be useful as a vector for vaccinating against other infectious agents as well. As an initial application, a live recombinant VZV expressing Epstein-Barr virus (EBV) membrane glycoproteins (gp350/220) was generated by inserting a gene fusion of the VZV gpI promoter and hydrophobic leader-encoding sequence with the gp350/220 coding sequence into the thymidine kinase (TK) gene of VZV (Oka). Insertion of the foreign DNA into the thymidine kinase gene was demonstrated by Southern blot analysis and the ability of the recombinant virus to replicate in the presence of bromodeoxyuridine. RNA splicing, glycosylation, and plasma membrane presentation of gp350/220 in cells infected with the recombinant virus were similar to those seen in EBV-infected cells. In addition, the expression of VZV-specific glycoproteins was unaltered by the concomitant expression of this large foreign glycoprotein. Thus, VZV can be used as a live viral vector for active immunization against EBV and other pathogens. PMID- 3035558 TI - The narL gene product activates the nitrate reductase operon and represses the fumarate reductase and trimethylamine N-oxide reductase operons in Escherichia coli. AB - Escherichia coli, which can utilize O2, nitrate, fumarate, or trimethylamine N oxide (Me3NO) as terminal electron acceptor, preferentially utilizes the one with the highest redox potential. Thus O2 prevents induction of nitrate, fumarate, and Me3NO reductases, and nitrate curtails the induction of fumarate and Me3NO reductases. Under anaerobic conditions the narL gene product, in the presence of nitrate, is known to activate transcription of the narC operon, which encodes nitrate reductase. This study shows that the same product plays a role in the repression by nitrate of the operons (frd and tor) that encode fumarate and Me3NO reductases. In contrast, the anaerobic repression of ethanol dehydrogenase by nitrate does not require the narL product. Expression of narL does not require the fnr gene product, a pleiotropic activator that is required for full expression of narC, frd, and tor. PMID- 3035559 TI - Use of 2-[125I]iodomelatonin to characterize melatonin binding sites in chicken retina. AB - 2-[125I]Iodomelatonin binds with high affinity to a site possessing the pharmacological characteristics of a melatonin receptor in chicken retinal membranes. The specific binding of 2-[125I]iodomelatonin is stable, saturable, and reversible. Saturation experiments indicated that 2-[125I]iodomelatonin labeled a single class of sites with an affinity constant (Kd) of 434 +/- 56 pM and a total number of binding sites (Bmax) of 74.0 +/- 13.6 fmol/mg of protein. The affinity constant obtained from kinetic analysis was in close agreement with that obtained in saturation experiments. Competition experiments showed a monophasic reduction of 2-[125I]iodomelatonin binding with a pharmacological order of indole amine affinities characteristic of a melatonin receptor: 2 iodomelatonin greater than 6-chloromelatonin greater than or equal to melatonin greater than or equal to 6,7-dichloro-2-methylmelatonin greater than 6 hydroxymelatonin greater than or equal to 6-methoxymelatonin much greater than N acetyltryptamine greater than N-acetyl-5-hydroxytryptamine greater than 5 methoxytryptamine greater than 5-hydroxytryptamine (inactive). The affinities of these melatonin analogs in competing for 2-[125I]iodomelatonin binding sites were correlated closely with their potencies for inhibition of the calcium-dependent release of [3H]dopamine from chicken and rabbit retinas, indicating association of the binding site with a functional response regulated by melatonin. The results indicate that 2-[125I]iodomelatonin is a selective, high-affinity radioligand for the identification and characterization of melatonin receptor sites. PMID- 3035560 TI - Poliovirus polypeptide precursors: expression in vitro and processing by exogenous 3C and 2A proteinases. AB - Plasmids have been constructed to generate substrates for the study of proteinases 2A and 3C of poliovirus. They contain the P1 (capsomer precursor) region of the poliovirus genome or P1 and part of P2 (a nonstructural precursor), which can be transcribed and translated in vitro. A transcript containing the entire 5' nontranslated region and the P1 region of the viral RNA gave poor translation in a reticulocyte translation system. Truncation of the 5' nontranslated region to its 3'-most segment gave acceptably good yields of radiolabeled P1. P1 was specifically processed to yield capsomer proteins by enzymes supplied in a postmitochondrial supernatant from poliovirus-infected cells. Thus, proteinase 3C can be supplied exogenously (in trans) and effect processing. This system may be used to provide P1 for the assay of proteinase 3C. Precursors that lacked either the 1A or 1D regions were poor substrates for proteinase 3C--observations that demonstrated a stringent structural requirement in processing by 3C. The translation product of a transcript encoding P1 and part of P2 was rapidly cleaved at the P1-P2 site in the absence of infected-cell extract. A transcript that contained a mutated 2A region gave a stable P1-P2 precursor that could be processed specifically by exogenous proteinase from infected-cell fractions. Processing of P1 appeared to require cleavage of the P1 P2 bond. These results support our previous data that 2A is the second polioviral proteinase and also provides a means of assaying proteinase 2A in vitro. PMID- 3035562 TI - Consensus topography in the ATP binding site of the simian virus 40 and polyomavirus large tumor antigens. AB - The location and sequence composition of a consensus element of the nucleotide binding site in both simian virus 40 (SV40) and polyomavirus (PyV) large tumor antigens (T antigens) can be predicted with the assistance of a computer-based pattern-matching system, ARIADNE. The latter was used to optimally align elements of T antigen primary sequence and predicted secondary structure with a "descriptor" for a mononucleotide binding fold. Additional consensus elements of the nucleotide binding site in these two proteins were derived from comparisons of T antigen primary and predicted secondary structures with x-ray structures of the nucleotide binding sites in four otherwise unrelated proteins. Each of these elements was predicted to be encompassed within a 110-residue segment that is highly conserved between the two T antigens residues 418-528 in SV40 T antigen and residues 565-675 in PyV). Results of biochemical and immunologic experiments on the nucleotide binding behavior of these proteins were found to be consistent with these predictions. Taken together, the latter have resulted in a topological model of the ATP binding site in these two oncogene products. PMID- 3035561 TI - cDNA and deduced amino acid sequence of human pulmonary surfactant-associated proteolipid SPL(Phe). AB - Hydrophobic surfactant-associated protein of Mr 6000-14,000 was isolated from ether/ethanol or chloroform/methanol extracts of mammalian pulmonary surfactant. Automated Edman degradation in a gas-phase sequencer showed the major N-terminus of the human low molecular weight protein to be Phe-Pro-Ile-Pro-Leu-Pro-Tyr-Cys Trp-Leu-Cys-Arg-Ala-Leu-. Because of the N-terminal phenylalanine, the surfactant protein was designated SPL(Phe). Antiserum generated against hydrophobic surfactant protein(s) from bovine pulmonary surfactant recognized protein of Mr 6000-14,000 in immunoblot analysis and was used to screen a lambda gt11 expression library constructed from adult human lung poly(A)+ RNA. This resulted in identification of a 1.4-kilobase cDNA clone that was shown to encode the N terminus of the surfactant polypeptide SPL(Phe) (Phe-Pro-Ile-Pro-Leu-Pro-) within an open reading frame for a larger protein. Expression of a fused beta galactosidase-SPL(Phe) gene in Escherichia coli yielded an immunoreactive Mr 34,000 fusion peptide. Hybrid-arrested translation with this cDNA and immunoprecipitation of [35S]methionine-labeled in vitro translation products of human poly(A)+ RNA with a surfactant polyclonal antibody resulted in identification of a Mr 40,000 precursor protein. Blot hybridization analysis of electrophoretically fractionated RNA from human lung detected a 2.0-kilobase RNA that was more abundant in adult lung than in fetal lung. The larger RNA and translation product indicates that SPL(Phe) is derived by proteolysis of a large polypeptide precursor. The amino acid sequence of the predicted protein, beginning Phe-Pro-Ile-Pro-Leu-Pro-Try-, comprises a hydrophobic peptide that is a major protein component of surfactant lipid extracts used successfully to treat hyaline membrane disease in newborn infants. These proteins, and specifically SPL(Phe), may therefore be useful for synthesis of replacement surfactants for treatment of hyaline membrane disease in newborn infants or of other surfactant deficient states. PMID- 3035564 TI - Rod phosphorylation favors folding in a catch muscle myosin. AB - Myosin from a molluscan catch muscle is unusual in being phosphorylated in the rod by an endogenous heavy chain kinase. The overall structure of the molecule resembles that of other muscle myosins, although the tail is somewhat longer (approximately equal to 1700 A). At low ionic strength the unphosphorylated molecules associate in filaments that display a striking axial repeat of 145 A. Phosphorylation of the rod enhances myosin solubility in the range of NaCl between 0.05 and 0.15 M. Depending on the ionic strength and the counterions present, the soluble species corresponds to an antiparallel folded dimer (15 S) or to a folded monomer (10 S). Unphosphorylated myosin can also be partially solubilized into folded monomers by addition of ATP in 0.15 M NaCl. A similar molecular folding has also been observed in smooth muscle and nonmuscle myosins that depends, however, on the state of phosphorylation of the light chains in the myosin head. We discuss these results in relation to possible mechanisms for control of catch contraction. PMID- 3035563 TI - Multiple genes encode the human Na+,K+-ATPase catalytic subunit. AB - A human genomic library was constructed and screened with hybridization probes derived from sheep and rat cDNAs encoding the alpha and alpha(+) isoforms, respectively, of the Na+,K+-ATPase catalytic subunit. Genomic sequences spanning 150 kilobases were isolated. Four genes, designated alpha A, alpha B, alpha C, and alpha D, each 20-25 kilobases in length, were identified by restriction mapping, Southern blot hybridization analysis, and limited DNA sequencing. We present evidence that two of these genes, alpha A and alpha B, encode the alpha and alpha(+) isoforms, respectively. The other genes, alpha C and alpha D, one of which is physically linked to the alpha(+) gene, exhibit nucleotide and amino acid homology to Na+,K+-ATPase catalytic subunit cDNA sequences but do not correspond to any previously identified isoforms. PMID- 3035566 TI - Expression of P1-450 and P3-450 DNA coding sequences as enzymatically active cytochromes P-450 in mammalian cells. AB - Two cDNA clones representing the mRNA coding sequences for mouse cytochromes P1 450 and P3-450 were inserted into the thymidine kinase gene of the wild-type vaccinia virus under the control of the vaccinia virus promoter. Murine and human cells infected with each of the resulting infectious recombinant viruses efficiently expressed their respective P-450 proteins. The newly synthesized protein products are translocated into the microsomes, and their characterization by immunochemical analysis indicates that the sizes of the polypeptides expressed were indistinguishable from their cytochrome P-450 counterparts found in mammalian liver microsomes. Functional analysis of each of the proteins by spectral and enzymatic analysis indicates that the expressed proteins have incorporated heme, and the holoenzymes displayed catalytic activities characteristic of their respective cytochrome P-450 enzymes. Thus, this system can be used to produce properly processed and catalytically active P-450 gene products in a wide variety of cells. The remarkable fidelity of expression and processing of these enzymes suggests that the vaccinia virus recombinants can be used for a wide variety of studies, including analysis of the effects of defined mutations produced in vitro, and directly correlate the structure/activity relationships of the cytochrome P-450 enzymes. PMID- 3035567 TI - Specific requirement for ATP at an early step of in vitro transcription of human mitochondrial DNA. AB - The ATP concentrations allowing transcription of both heavy- and light-strand of human mtDNA in a HeLa cell mitochondrial lysate were found to cover a broad range, with a maximum around 2.5 mM, and with reproducible differences in the ATP response curves for the two transcription events. Direct measurements showed that nonspecific ATP degradation during the assay did not account for the high ATP requirement. 5'-Adenylyl imidodiphosphate (p[NH]ppA), an ATP analog with a nonhydrolyzable beta-gamma bond, was unable to substitute for ATP in supporting mtDNA transcription but greatly stimulated this transcription in the presence of a low concentration of exogenous ATP. Evidence was obtained indicating that p[NH]ppA did not support an early event in mtDNA transcription (formation of preinitiation complex or initiation), whereas this analog could substitute effectively for ATP in the subsequent elongation steps. These results pointed to a specific requirement for ATP at an early step of the transcription process. PMID- 3035568 TI - Synthetic peptide antagonists of glucagon. AB - Several glucagon analogs were synthesized in an effort to find derivatives that would bind with high affinity to the glucagon receptor of rat liver membranes but would not activate membrane-bound adenylate cyclase and, therefore, would serve as antagonists of the hormone. Measurements on a series of glucagon/secretin hybrids indicated that replacement of Asp9 in glucagon by Glu9, found in secretin, was the important sequence difference in the N terminus of the two hormones. Further deletion of His1 and introduction of a C-terminal amide resulted in des-His1-[Glu9]glucagon amide, which had a 40% binding affinity relative to that of native glucagon but caused no detectable adenylate cyclase activation in the rat liver membrane. This antagonist completely inhibited the effect of a concentration of glucagon that alone gave a full agonist response. It had an inhibition index of 12. The pA2 was 7.2. An attempt was made to relate conformation with receptor binding. The peptides were synthesized by solid-phase methods and purified to homogeneity by reverse-phase high-performance liquid chromatography on C18-silica columns. PMID- 3035565 TI - Nucleotide sequence of medium-chain acyl-CoA dehydrogenase mRNA and its expression in enzyme-deficient human tissue. AB - Medium-chain acyl-CoA dehydrogenase (MCAD; acyl-CoA: (acceptor) 2,3 oxidoreductase, EC 1.3.99.3) is one of three similar enzymes that catalyze the initial step of fatty acid beta-oxidation. Definition of the primary structure of MCAD and the tissue distribution of its mRNA is of biochemical and clinical importance because of the recent recognition of inherited MCAD deficiency in humans. The MCAD mRNA nucleotide sequence was determined from two overlapping cDNA clones isolated from human liver and placental cDNA libraries, respectively. The MCAD mRNA includes a 1263-base-pair coding region and a 738-base-pair 3' nontranslated region. A partial amino acid sequence (137 residues) determined on peptides derived from MCAD purified from porcine liver confirmed the identity of the cDNA clone. Comparison of the amino acid sequence predicted from the human MCAD cDNA with the partial protein sequence of the porcine MCAD revealed a high degree (88%) of interspecies sequence identity. RNA blot analysis shows that MCAD mRNA is expressed in a variety of rat (2.2 kilobases) and human (2.4 kilobases) tissues. Blot hybridization of RNA prepared from cultured skin fibroblasts from a patient with MCAD deficiency disclosed that mRNA was present and of similar size to MCAD mRNA derived from control fibroblasts. The isolation and characterization of MCAD cDNA is an important step in the definition of the defect underlying MCAD deficiency and in understanding its metabolic consequences. PMID- 3035570 TI - Specific binding of erythropoietin to its receptor on responsive mouse erythroleukemia cells. AB - Erythropoietin (Epo) is a glycoprotein factor that specifically regulates the proliferation and differentiation of erythroid progenitor cells. Here we describe the isolation of Epo-responsive mouse erythroleukemia cell line SKT6, the characterization of the specific binding of biologically active 125I-labeled human Epo (125I-Epo) to its membrane receptor, and, finally, report information concerning the molecular structure of the receptor. About 75% of erythroid colony forming precursor cell-like colonies derived from SKT6 cells were hemoglobin positive after 3- to 4-day exposure to Epo in methylcellulose culture. Radioiodinated Epo bound specifically to SKT6 cells, and Scatchard analysis of the data showed a high affinity for 125I-Epo (Kd = 0.15 nM) but displayed only a small number of specific receptors (approximately equal to 470 per cell). Membrane components that specifically interact with 125I-Epo were identified by covalent crosslinking with disuccinimidyl suberate, and three receptor species with apparent Mr 63,000, 94,000, and 119,000 were found in membrane from SKT6 cells, suggesting the complex structure of the receptor molecules. Specific bindings were also detected in all of the Epo-unresponsive Friend erythroleukemia cells examined, and cross-linking study revealed the presence of only the 63,000 species as a binding site. PMID- 3035569 TI - Deuterium nuclear magnetic resonance measurements of blood flow and tissue perfusion employing 2H2O as a freely diffusible tracer. AB - The use of deuterium oxide (2H2O) is proposed as a freely diffusible nuclear magnetic resonance (NMR) blood flow and tissue perfusion tracer of potential clinical utility. Deuterium is a stable, nonradiative isotope commercially available as 2H2O at enrichment levels of essentially 100%--i.e., 110 molar equivalent deuterium. This high concentration, together with the short relaxation time of the spin 1 (quadrupole) deuterium nuclide, provides substantial sensitivity for NMR spectroscopy. As a result, when 2H2O is administered in a bolus fashion to a specific tissue or organ in vivo, the deuterium NMR intensity time course can be analyzed, using mathematical models developed by others for radiolabeled tracers, to measure the rate of blood flow and tissue perfusion. Such an application is demonstrated herein at a static magnetic field of 8.5 tesla. Using single-compartment flow modeling, hepatic blood flow and tissue perfusion in fasted (18 hr) male Sprague-Dawley rats was determined to be 61 +/- 17 (mean +/- SD) ml/100 g per min (n = 5). PMID- 3035571 TI - SSC1, a member of the 70-kDa heat shock protein multigene family of Saccharomyces cerevisiae, is essential for growth. AB - The genome of the yeast Saccharomyces cerevisiae contains a family of genes related to the HSP70 genes (encoding the 70-kDa heat shock protein) of other eukaryotes. Mutations in two of these yeast genes (SSC1 and SSD1), whose expression is increased a few fold after temperature upshift, were constructed in vitro and substituted into the yeast genome in place of the wild-type alleles. No phenotypic effects of the mutation in SSD1 were detected. However, a functional SSC1 gene is essential for vegetative growth. This result, in conjunction with experiments involving mutations in other members of this multigene family, indicates that at least three distinct functions are carried out by genes of the HSP70 family. PMID- 3035572 TI - Microcell-mediated transfer of a single human chromosome complements xeroderma pigmentosum group A fibroblasts. AB - Chromosomes from an immortalized aneuploid human fibroblast cell line were randomly tagged with the selectable marker neo by transfection with the plasmid pSV2neo. Somatic cell fusions between transfected human cells and mouse A9 cells generated pools of G418-resistant human-mouse hybrid clones containing various numbers of human chromosomes. Microcell-mediated chromosome transfer from the hybrid pools to xeroderma pigmentosum complementation group A (XP-A) cells in culture and selection for G418-resistant colonies resulted in the identification of XP cells with enhanced resistance to ultraviolet radiation. Screening of subclones from selected pools of human-mouse hybrids facilitated the identification of hybrids containing a single neo-tagged human chromosome. Transfer of this chromosome to XP-A cells (but not to XP-F or XP-C cells) results in enhanced resistance to ultraviolet light and enhanced excision repair capacity. The identification of a single human chromosome that complements the phenotype of XP-A cells in culture provides the potential for genetic mapping of the complementing gene and for its isolation by molecular cloning. PMID- 3035573 TI - Modulation of transcription by DNA supercoiling: a deletion analysis of the Escherichia coli gyrA and gyrB promoters. AB - Expression of the genes determining the subunits of Escherichia coli DNA gyrase (gyrA and gyrB) is known to be induced by relaxation of the template DNA. In this paper we report a deletion analysis of the gyrA and gyrB promoter regions. We find that a DNA sequence 20 base pairs long that includes the -10 consensus region, the transcription start point, and the first few transcribed bases is responsible for the property of induction by DNA relaxation. We propose a model for relaxation-stimulated transcription in which promoter clearance is the rate limiting step. PMID- 3035574 TI - Molecular cloning and characterization of a cDNA encoding the cerebrovascular and the neuritic plaque amyloid peptides. AB - Deposits of amyloid fibers are found in large numbers in the walls of blood vessels and in neuritic plaques in the brains of patients with Alzheimer disease and adults with Down syndrome. We used the amino acid sequence of the amyloid peptide to synthesize oligonucleotide probes specific for the gene encoding this peptide. When a human brain cDNA library was screened with this probe, a clone was found with a 1.7-kilobase insert that contains a long open reading frame coding for 412 amino acid residues including the 28 amino acids of the amyloid peptide. RNA gel blots revealed that a 3.3-kilobase mRNA species was present in the brains of individuals with Alzheimer disease, with Down syndrome, or with no apparent neurological disorders. Southern blots showed that homologous genes are present in the genomic DNA of humans, rabbits, sheep, hamsters, and mice, suggesting that this gene has been conserved through mammalian evolution. Localization of the corresponding genomic sequences on human chromosome 21 suggests a genetic relationship between Alzheimer disease and Down syndrome, and it may explain the early appearance of large numbers of neuritic plaques in adult Down syndrome patients. PMID- 3035575 TI - Molecular cloning of Lyt-3, a membrane glycoprotein marking a subset of mouse T lymphocytes: molecular homology to immunoglobulin and T-cell receptor variable and joining regions. AB - Lyt-3 is a membrane glycoprotein expressed on thymocytes and class I major histocompatibility complex-restricted cytotoxic T cells. Lyt-3 is expressed as a heterodimer with Lyt-2, and this complex is considered to be a homologue of the human Leu-2/T8 (CD8) that has been postulated to be a receptor for the class I major histocompatibility complex. We have determined the complete primary structure of Lyt-3 from the nucleotide sequence of its cDNA clones. Analysis of the predicted amino acid sequence indicates that the Lyt-3 polypeptide has a 21 amino acid leader peptide, and the mature protein consists of an NH2-terminal region of 146 amino acids, a transmembrane region of 27 residues, and a C terminal region of 19 amino acids. The NH2-terminal 110 residues show clear homology to the T-cell receptor and immunoglobulin variable region sequences. In addition, Lyt-3 has 11 residues that have strong homology to the joining region sequences of the T-cell receptor and the immunoglobulin heavy and light chains. The presence of immunoglobulin variable- as well as joining-region-related sequences in Lyt-3 further supports the idea that these molecules may be recognition molecules belonging to the immunoglobulin super gene family. PMID- 3035576 TI - DNA segment containing C beta 1, a gene for the constant region of the beta chain of the T-cell antigen receptor, was inserted into chromosome 6 in cells from one patient with human T-cell leukemia. AB - DNA rearrangements that occurred in the vicinity of T-cell antigen receptor beta chain gene clusters residing on chromosome 7 were examined in human T-cell acute lymphoblastic leukemia cells. In one patient, it was observed that, for the T cell receptor beta-chain genes, a D beta 1-J beta 2.3 (where D is diversity and J is joining) junction was found on one chromosome, while the other chromosome kept the germ-line configuration. If this D beta-J beta junction was formed by the customary deletion mechanism, the C beta 1 gene (where C is constant) located between the D beta 1 and J beta 2.3 loci should have disappeared from this chromosome. The C beta 1 gene indeed was absent from the rearranged chromosome 7, but it was found on chromosome 6 as an inserted segment. The implications of the observations are discussed. PMID- 3035577 TI - Two efficient ribosomal frameshifting events are required for synthesis of mouse mammary tumor virus gag-related polyproteins. AB - The primary translation products of retroviral pol genes are polyproteins initiated in an upstream gene (gag). To investigate the manner in which the gag initiated polyproteins of the mouse mammary tumor virus are produced, we determined the nucleotide sequence of a 1.8-kilobase DNA fragment that spans the region between gag and pol in the C3H strain of mouse mammary tumor virus. The sequence reveals three overlapping open reading frames: the first encodes products of gag (p27gag and p14gag); the second encodes a protein domain of unknown function (termed X) that is highly related to a similarly positioned sequence in simian type D retroviruses and the viral protease (pro); and the third encodes the reverse transcriptase. The reading frames are organized to permit uninterrupted readthrough from gag to pol if ribosomal frameshifts occur in the -1 direction within each of the two overlapping regions, one of which is 16 nucleotides in length and the other 13 nucleotides. Cell-free translation of RNA containing these overlap regions shows that fusion of the reading frames by ribosomal frameshifting occurs efficiently: about one-fourth of the ribosomes traversing the gag-X/pro overlap and one-tenth traversing the X/pro-pol overlap shift frames, generating gag-related polyproteins in ratios similar to those observed in vivo. Synthetic oligonucleotides containing either of the overlap regions inserted into novel contexts do not induce frameshifting; hence the overlapping portions of the reading frames are not sufficient to induce a frameshift event, and a larger sequence context or secondary structure may be implicated. PMID- 3035578 TI - Identification of a herpes simplex virus 1 glycoprotein gene within a gene cluster dispensable for growth in cell culture. AB - The genome of herpes simplex virus 1 consists of two components, L and S, each containing unique sequences flanked by inverted repeats. Current and earlier studies have shown that 11 of the 12 open reading frames contained in the unique sequences of the S component can be deleted and are dispensable for growth in cell culture. Analyses of one recombinant virus containing a deletion in the open reading frame US7 permitted the identification of a monoclonal antibody specific for the product of this gene. The protein encoded by this gene has a predicted translated molecular weight of 41,366 and an apparent molecular weight of approximately 65,000 in denaturing polyacrylamide gels. The electrophoretic mobility of the protein synthesized by cells in the presence of inhibitory concentrations of tunicamycin is faster than that of the protein accumulating in lysates of untreated infected cells. We conclude that the product of US7 is glycoprotein subject to N-linked glycosylation, and we have designated it glycoprotein I. These studies indicate that the unique sequences of the S component encode four glycoproteins (G, D, I, and E) of which at least three (G, I, and E) are dispensable for growth in continuous lines of primate cells. PMID- 3035580 TI - The cardiac anti-adrenergic effect of adenosine. AB - Adenosine has an anti-adrenergic action in the heart and appears to serve as negative-feedback modulator of beta-adrenoceptor-mediated myocardial contractile and metabolic responses. This action of adenosine has been assessed in various heart preparations by the application of exogenous adenosine, the determination of tissue and coronary venous effluent levels of adenosine and the degradation of endogenous adenosine by adenosine deaminase. The evidence suggests that the anti adrenergic function of adenosine has vital physiological and clinical implications in the normoxic, hypoxic and ischemic myocardium. PMID- 3035579 TI - Neuroanatomical patterns of the mu, delta, and kappa opioid receptors of rat brain as determined by quantitative in vitro autoradiography. AB - Highly specific radioligands and quantitative autoradiography reveal strikingly different neuroanatomical patterns for the mu, delta, and kappa opioid receptors of rat brain. The mu receptors are most densely localized in patches in the striatum, layers I and III of the cortex, the pyramidal cell layer of the hippocampal formation, specific nuclei of the thalamus, the pars reticulata of the substantia nigra, the interpeduncular nucleus, and the locus coeruleus. In contrast, delta receptors are highly confined, exhibiting selective localization in layers I, II, and VIa of the neocortex, a diffuse pattern in the striatum, and moderate concentration in the pars reticulata of the substantia nigra and in the interpeduncular nucleus. delta receptors are absent in most other brain structures. This distribution is unexpected in that the enkephalins, the putative endogenous ligands of the delta receptor, occur essentially throughout the brain. The kappa receptors of rat brain exhibit a third pattern distinct from that of the mu and delta receptors. kappa receptors occur at low density in patches in the striatum and at particularly high density in the nucleus accumbens, along the pyramidal and molecular layers of the hippocampus, in the granular cell layer of the dentate gyrus, specific midline nuclei of the thalamus, and hindbrain regions. kappa receptors appear to be uniformly distributed across regions in the neocortex with the exception of layer III, which revealed only trace levels of binding. An important conclusion of the present study is that delta receptors occur at high density only in the forebrain and in two midbrain structures, whereas mu and kappa receptors exhibit discrete patterns in most major brain regions. PMID- 3035581 TI - Removal of Limulus test-interfering factors in blood samples with perchloric acid and the improvement of the specificity of the Limulus test by fractionating amebocyte lysate. PMID- 3035582 TI - Recovery of endotoxin from human plasma by acid oxidative treatments as monitored by an automated microtiter plate-chromogenic substrate Limulus amebocyte lysate (LAL) assay method. AB - Methods to detect endotoxin in human plasma for LAL testing have been developed by use of a perchloric acid (PCA) or a trifluoroacetic acid (TFA) treatment. These acid oxidative treatments eliminate interferences present in plasma. The mode of action of the acids for the elimination of the interferences is indicated to be by their oxidative actions. The treated plasma is assayed by use of an automated microtiter plate-chromogenic substrate LAL assay method employing a modified Cetus Pro/Pette EXPRESS system. The automated system assays 96 samples in approximately 22 minutes. Assay sensitivity of the automated Pro/Pette system is approximately 0.01 endotoxin units (EU) per ml with a linear range from 0.03 to 0.14 EU/ml. The relative standard deviation (RSD) of the assay system is approximately 6%. The improved precision of the LAL assay by the automated system has been attributed to its fast speed (2 minutes) in dispensing the unstable LAL reagent to all 96 samples in one assay block. The total operation, including the acid oxidative treatments and the 96 sample LAL assay, takes approximately 45 minutes. The recovery of spiked endotoxin from plasma ranged from 75 to approximately 100% by adding 0.4 ml of either 0.1-0.2 N PCA or 0.2-0.35 N TFA to 0.2 ml plasma and reacting for 15 minutes at 37 degrees C. The standard deviation of the assay is approximately 0.1 EU/ml plasma. The method has been proven to be rugged, simple, rapid, and cost effective thus would be suitable for clinical application. PMID- 3035583 TI - Binding studies of dermorphin and its L-form on rat brain opioid receptors. AB - It is well known dermorphin is a potent and long-acting opioid peptide while its synthetic L-form is almost completely devoid of biological activity. We investigated whether the L-Ala2 residue might affect the affinity of the compound for opioid receptors or make [L-Ala2] dermorphin more sensitive to metabolic degradation. Dermorphin and [L-Ala2] dermorphin were assayed in [3H]naloxone binding to opioid receptors in rat brain preparations in the absence and presence of peptidase inhibitors bestatin, captopril and thiorphan. The synthetic [L-Ala2] dermorphin showed very low affinity for the opioid receptors. This was only slightly increased in the presence of the peptidase inhibitor bestatin, alone and in combination with captopril and thiorphan. The low affinity of [L-Ala2] dermorphin was not improved even when the binding assay was carried out at 0 degrees C. We suggest that the D-Ala2 residue is essential for the binding of dermorphin to the opioid receptors as well as for its pharmacological activity. PMID- 3035584 TI - Receptors in the gastrointestinal tract. AB - The receptor concept has been recently evolved and a new science was actually created, namely "receptorology". Receptors are now identified by means of different techniques (binding, agonist-antagonist interaction, autoradiography, etc.). The new techniques allowed the investigators to define new receptors and new subtypes of the "classical" ones. In the gastrointestinal (GI) tract a number of receptors have been identified and localized both on the effector organ and in the nerve terminal where they exert an important modulatory function on the neurotransmitter release. Recent biochemical studies have allowed a better understanding of the post-receptor event involving the second or third messenger regulation. Particular changes of receptors were recognized and they allow us to consider receptors not as static entities but as very dynamic components of the plasma membrane capable of different kinds of alterations, like interconversion, internalization, mobility, up- and downregulation, etc. Together with the "classical" receptors (cholinergic, adrenergic, opioid, etc.) also new receptors were identified: different subtypes of receptors for the tachykinins, for prostaglandin of the E type in the gastric parietal cell and the so-called dihydropyridine (DHP) receptor in the calcium channel of different areas of the gut. It is obvious that the precise knowledge of receptors and of their agonists and antagonists will represent the basis for a more specific and efficacious treatment of various gastrointestinal disorders. PMID- 3035586 TI - Decreased gastro-intestinal responses to certain agonists in streptozotocin- and alloxan-diabetic rats in vitro. AB - In this study, the beta-adrenergic, muscarinic and serotonergic receptor activities were investigated in alloxan- and streptozotocin-diabetic rats using duodenum, ileum and gastric fundus strips as assay organs. In addition, the effect of manganese chloride on the duodenum of normal and diabetic rats was studied to understand whether it affects the gastro-intestinal adenylate cyclase activity. The results obtained in this study suggest that no significant changes occur in the gastro-intestinal muscarinic receptor and adenylate cyclase activities due to experimental diabetes. Nevertheless, the experiments performed on the rat duodenum and gastric fundus strips with salbutamol and serotonin showed that the beta-adrenergic and serotonergic receptor activities of the gastro-intestinal tract decreased significantly in alloxan- and streptozotocin induced diabetes. The results obtained seem to show that the gastro-intestinal complication(s) of the diabetes were related to the reduction(s) of the beta adrenergic and/or serotonergic receptor activities. PMID- 3035585 TI - Renal catecholamines and alpha 2-adrenergic receptors in salt-related and genetic hypertension. AB - Increased dietary salt intake alters renal function which often leads to deleterious cardiovascular consequences. Studies were carried out to characterize the effects of high-salt diets on renal catecholamines and alpha 2-adrenergic receptors. These parameters were evaluated in both genetic and acquired forms of hypertension and also in normotensive rats on high-salt diets. Renal catecholamine content was determined by high-performance liquid chromatography with electrochemical detection. Renal alpha 2-adrenergic receptor-binding studies were performed on whole kidney homogenates using 3H-p-aminoclonidine to label both high- (0.5 nM) and low-affinity (5.0 nM) renal alpha 2-adrenergic receptors. Increased salt intake elevated blood pressure, decreased renal norepinephrine stores and resulted in renal alpha 2-adrenergic receptor up-regulation in deoxycorticosterone acetate salt hypertensive rats, Dahl-S rats and COX-SHR. The decreased renal stores of norepinephrine (NE) appeared to reflect increased renal NE utilization. In contrast, SHR (Charles River) had elevated NE stores and alpha 2-adrenergic receptors while on normal salt diets. Short-term (10-14 days) exposure to high-salt diets had modest effects in normotensive rats or COX-SHR, although it was sufficient to increase low affinity renal alpha 2-adrenergic receptor number. Renal dopamine metabolism was also altered by high-salt diets. These studies demonstrated a relationship between renal NE content and renal alpha 2-adrenergic receptors. The implications of this relationship and other salt-related changes in renal catecholamine metabolism were discussed as they pertained to hypertension and renal function. PMID- 3035587 TI - Effects of delta opioid antagonists on enkephalin-induced seizures. AB - We examined the effect of opioid receptor antagonists on the seizure phenomena induced by specific delta opioid receptor agonist [D-Ser2,Leu5] enkephalyl-Thr (DSLET). The experiments have been performed in the anesthetized rats, and the DSLET-induced seizure phenomena were registered by electrocorticogram and electromyogram. It was demonstrated that two selective delta opioid receptor antagonists, ICI 152,129 and ICI 174,864 inhibited DSLET-induced epileptiform ECoG pattern and myoclonic contractions in a dose-related manner. An equimolar concentration of naloxone failed to antagonize the epileptiform effects of DSLET. It is concluded that delta opioid receptor agonist-induced seizure is mediated by delta receptors, since it can be blocked by delta opioid receptor antagonists. Evidently, delta opioid antagonists can be used as a good tool, in order to demonstrate a delta component in the seizure phenomena induced by other endogenous opioid peptides or their derivatives. PMID- 3035588 TI - Inhibition of ribonucleotide reductases encoded by herpes simplex viruses. PMID- 3035589 TI - Manipulating the dopaminergic system in Parkinson's disease. PMID- 3035590 TI - Is there a common denominator for the antimanic effect of lithium and anticonvulsants? AB - Starting from the idea that different primary actions of carbamazepine, valproate and lithium might converge into a similar secondary mechanism of action, e.g. a modulation of the functioning of certain transmitter systems, a review of the literature was made on the effects of these and related substances on GABAergic, dopaminergic, cholinergic, noradrenergic and serotonergic transmission, in an attempt to find a common denominator. It was concluded that similarities of the effects of these drugs are most evident with respect to a reduction of dopaminergic transmission. Some similarities also exist with respect to GABAergic and, to a lesser degree, cholinergic transmission. The effects on noradrenergic and serotonergic transmission seem too disparate to be considered as a potential basis for the antimanic effects of these drugs. PMID- 3035591 TI - Dielectric, haematological and biochemical studies of detergent toxicity in fish blood. AB - Blood characteristics in dielectric, haematological and biochemical terms of the fish Cyprinus carpio exposed to a sublethal concentration of sodium alkyl benzene sulphonate were compared with those from untreated control fish; recovery from the test solution was also checked. Trends of change in the majority of chosen parameters of blood with time of fish exposure to anionic detergent were significantly linear. A decline was noted in erythrocyte count, haematocrit, blood haemoglobin concentration, mean corpuscular haemoglobin concentration and mean corpuscular haemoglobin with the exception of mean cellular volume. An uptake of sodium into red blood cells and a rise of intracellular potassium were seen. Practical indicators of the presence of detergent in fish blood were obtained from dielectric beta dispersion measurements. It has been shown that dielectric parameters could be correlated with haematological parameters but not with a biochemical one. Results indicate that the changes in haematocrit induce corresponding variation of the maximum of the dielectric loss factor tan delta. An increase in the electrical conductivity and permittivity of the erythrocytes' interior in exposed fish was associated with a decline of mean corpuscular haemoglobin. PMID- 3035593 TI - [Does cellulose treat cancer?]. PMID- 3035592 TI - The topography and dynamics of cholinergic transmission in the myenteric plexus smooth muscle preparation of the guinea-pig ileum; the effect of ketocyclazocine, a kappa opiate ligand. AB - Output of acetylcholine (ACh), neurogenic electromyogram (NEMG) and contractions of guinea-pig ileum preparations were studied during stimulation by high frequency trains of impulses. Under control conditions the output of ACh per impulse after 2nd to 4th impulses during train stimulation (30 Hz) was higher by 20-40% than the level of ACh output during the first impulse. In the presence of ketocyclazocine (KTZ, 80 nmol x l-1) the output of ACh evoked by the first impulse was more effectively inhibited than that after impulses 2 to 4 so that the increase was higher (80-170%). NEMG, a direct consequence of the localized action of released transmitter (ACh), was recorded in the longitudinal muscle 4 and 10 mm aborally from the focal stimulation site. The incidence of NEMG responses was higher at the proximal than at the distal site and was proportional to the number of impulses in a train (100 Hz). At the distal site KTZ suppressed the appearance of NEMG responses to single impulses whereas at the proximal site its effect was much less; and so was its effect at either site during train stimulation. It is concluded that in the course of train stimulation, sites of transmission more distant from the stimulation focus were recruited, and consequently the secretion of ACh in succeeding impulses was enhanced. KTZ might preferentially inhibit the propagation of excitation by the very first impulse. PMID- 3035594 TI - [Angiotensin converting enzyme activity in smokers]. PMID- 3035595 TI - [Myoblastoma granulare of the bronchi]. PMID- 3035596 TI - [The value of cytologic typing of bronchial carcinoma]. PMID- 3035597 TI - Beta-adrenoceptors on lymphocytes of patients with major depressive disorder. AB - 3H-Dihydroalprenolol binding to lymphocyte membranes of patients with primary, unipolar major depressive disorder was compared to that of a normal, healthy control population. No significant difference could be demonstrated between the Kd values of the two different groups, but the Bmax values of the depressed patients were significantly lower than those of the controls. Positive correlations were observed between the lymphocyte beta-adrenoceptor Bmax values of the patients and their Beck self-evaluation and Hamilton depression ratings. We propose that decreased lymphocyte beta-adrenoceptor Bmax values may be used as a biological marker for major depressive disorder. PMID- 3035598 TI - The effects of the systemic administration of N-methylmorphine chloride, a quaternary analogue of morphine that does not cross the blood-brain barrier, on the release of anterior pituitary hormones in the rat. AB - The acute administration of morphine elicits changes in the release of anterior pituitary hormones. The locus of this action is thought to be in the central nervous system, specifically the hypothalamus. There are some data suggesting that systemically administered opiates and opioid peptides can act outside of the blood-brain barrier to influence anterior pituitary hormone release. To test this hypothesis we examined the effects of the systemic administration of N methylmorphine chloride, a quaternary analogue of morphine that does not cross the blood-brain barrier, on the release of corticosterone, growth hormone, prolactin, luteinizing hormone, and thyroid stimulating hormone in the rat. N methylmorphine caused increases in the release of growth hormone and prolactin, but serum levels of corticosterone, luteinizing hormone and thyroid stimulating hormone were unaffected. These results indicate that the opiate-induced release of growth hormone and prolactin may be mediated in part by sites outside of the blood-brain barrier. PMID- 3035599 TI - Transient changes in permeability in HeLa and L cells during detachment from a substrate. AB - The Na+ and K+ gradients of HeLa cells approach that of the medium during removal from a substrate with trypsin, but then recover during the next 15 min. The recovery is blocked by ouabain or the cold, but is unaffected by bumetanide. The effect is also obtained in cells which have no intercellular connexions, and in cells whose interior is made acid with CO2. Removal of cells with EDTA, pronase E and dispase has similar effects. It does not occur, or is greatly reduced, in cells already rounded up. Substances of molecular weight up to 5000 (e.g. inulin) also cross the cell membrane during this phase. We think that the effect is due to a transient increase in leakiness of the cell during rounding up, possibly due to the detachment of the 'feet' holding the cells onto the substrate. The transient increase in permeability of these cells may be a valuable method of introducing large molecules into them. PMID- 3035600 TI - [Usual transplantation procedures and new implant materials in periodontal bone pocket surgery--an overview]. PMID- 3035601 TI - [Hydroxylapatite in the surgical therapy of advanced periodontal disease]. PMID- 3035602 TI - Free radicals from single crystals of deoxyguanosine 5'-monophosphate (Na salt) irradiated at low temperatures. AB - In single crystals of the DNA nucleotide 2'deoxyguanosine-5'phosphate (5'dGMP) X- or gamma-irradiated at 4.2 K or 15 K, two primary radical species can be discriminated and assigned to the cation and anion of the guanine base, G(+) and G(-). Both species are unstable. G(-) partially transforms into a secondary radical at 4.2 K, the latter being the precursor to the dominant 300 K species formed by net H-addition to carbon C8. The secondary radical, together with another intermediate appearing at 77 K and perhaps connected with the anion decay could not be structurally identified. The guanine cation G(+) transforms upon annealing to temperatures above 77 K into a more stable species by deprotonation at position N1. PMID- 3035604 TI - Application of overpressured layer chromatography in the quality control of radiopharmaceuticals. PMID- 3035603 TI - Effect of gamma irradiation on membranes of normal and pathological erythrocytes (beta-thalassemia). AB - The influence of ionizing radiation on the membrane of human normal erythrocytes has extensively been studied and a variety of effects including changes in the cation fluxes [3, 9] or in non-electrolytes permeability [5, 6, 11], in membrane fluidity, in peroxidation of unsaturated lipids as well as chemical composition or structural modifications [4, 7, 8] has been observed. However, only few studies deal with the effects of ionizing radiation on pathological red blood cells. In this work, we have investigated by means of electron spin resonance (ESR) spectroscopy the effects of 60Co gamma-radiation on the normal and homozygous beta-thalassemic human erythrocyte membranes [12, 13]. PMID- 3035605 TI - Hepatoblastoma: radiologic-pathologic correlation in 50 cases. AB - Fifty cases of hepatoblastoma were reviewed. Virtually all patients were infants or young children with hepatomegaly or a mass. Calcification seen on 11 of 20 radiographs was often in a pattern of small chunks, and eight of these correlated with osteoid formation in histologically mixed hepatoblastomas. Angiography generally showed tumor vascularity, sometimes with a spoke-wheel pattern. Suggestion of tumor nodularity or lobulation on sonography or computed tomography (CT) correlated with the gross appearance. The tumor was usually echogenic and occasionally had small hypoechoic or anechoic areas representing necrosis or hemorrhage. On CT it was usually hypodense, with minimal if any enhancement. The calcification pattern and demonstration of tumor lobulation with septation may help differentiate hepatoblastoma from other liver neoplasms in infants and children under 5 years of age. PMID- 3035606 TI - Hepatocellular carcinoma: MR imaging. AB - Sixty patients with hepatocellular carcinoma (HCC) were studied with computed tomography (CT) and magnetic resonance (MR) imaging at 1.5 T. MR imaging was equivalent to CT in detection of HCC. MR imaging was superior to CT in demonstrating the details of tumors, especially pseudocapsules. In 58 cases, main tumors were detected with MR imaging. On spin-echo (SE) 600/25 (repetition time msec/echo time msec) sequences, tumors were hyperintense in 18 cases, isointense in ten, and hypointense in 30. On SE 2,000/60 sequences, all but two tumors had high signal intensity. Pseudocapsules, intratumoral septa, daughter nodules, and tumor thrombi, which are important characteristics of HCC, were demonstrated in 22, three, six, and six cases, respectively, on MR imaging. MR imaging is useful for characterizing the internal architecture of HCC. PMID- 3035607 TI - Postoperative radiation therapy in the management of lung cancer. AB - Postoperative radiation therapy for lung cancer is still controversial. In a 9 year period, 69 patients with non-oat-cell carcinoma of the lung (16% stage I, 26% stage II, and 58% stage III) received such therapy. The radiation dose was less than 5,000 cGy in 42 patients, 5,000-5,900 cGy in 16, and 6,000 cGy or more in 11; follow-up ranged from 24 to 64 months. Actuarial survival at 2 and 4 years was 50% and 16%, respectively, for squamous cell carcinoma, and 40% and 26% for adenocarcinoma. The 5-year survival for stages I, II, and III cancer was 29%, 17%, and 19%, respectively. Histologic findings and type of surgery did not affect survival, but the radiation dose apparently did. The 3-year survival for patients who received less than 6,000 cGy was 35%, compared with 73% for patients who received higher doses. In eight patients, treatment failed within the irradiated volume: all had received doses of less than 6,000 cGy, and the volume in three was judged to be inadequate. PMID- 3035608 TI - Myelinated and nonmyelinated nerves: comparison of proton MR properties. AB - The magnetic resonance (MR) relaxation rates of protons were compared in the myelinated and nonmyelinated nerves of the garfish. The long, large olfactory nerve of the garfish, as an easily accessible source of nonmyelinated axons, is uniquely suited for such a comparison. The T1 and T2 measurements revealed distinct and consistent differences between nonmyelinated olfactory nerves and myelinated optic and oculomotor nerves. Comparisons between water content, lipid content, and relaxation rates indicated that the differences in MR properties represent complex differences in the distribution and physical environment of the constituent lipid and water protons. PMID- 3035609 TI - In vivo quantitation using SPECT of radiopharmaceutical uptake by human meningiomas. AB - A single photon emission computed tomographic method was designed for the measurement of radiopharmaceutical uptake in brain tumors. Results of phantom studies showed a correlation coefficient of .99 when measured volume was compared with actual volume. The correlation coefficient for measured radioactivity concentration compared with the actual concentration was .97. In 13 meningiomas the correlation between in vivo SPECT measurements of uptake and in vitro measurements in samples of the same tumors removed surgically was .84; when two tumors that contained regions of necrosis and fibrosis were excluded it was .93. This method can be used for in vivo quantitative assessment of pharmacokinetics of labeled drug uptake in human brain tumors. PMID- 3035610 TI - Agrin. PMID- 3035611 TI - The effect of drugs on oligodendrocyte proliferation and myelin regeneration. PMID- 3035613 TI - A simple and sensitive radioreceptor assay for leukotrienes. AB - A simple and sensitive radioreceptor assay (RRA) for leukotrienes (LTs) was developed using a highly specific [3H]leukotriene D4 (LTD4) binding to guinea pig lung membrane homogenates. The assay can detect down to 0.15 pmol of LTD4. The values for fifty percent inhibition of bound [3H]LTD4 was 1.5 nM for LTD4, 45 nM for LTC4 and 24 nM for LTE4. LTB4 at 3.0 X 10(-5)M had no effect on [3H]LTD4 binding. The RRA for LTs in the absence of serine-borate complex was bi-specific for both LTC4 and LTD4. However, in the presence of 20 mM serine-borate this method was highly specific for LTD4. Recovery rate averaged 87.2% after ethanol extraction and evaporation of known amounts of LTD4. When the radioreceptor assay and radioimmunoassay data for leukotriene levels in the samples were compared to each other, an excellent correlation was observed with a correlation coefficient 'r' of 0.992. The assay was also validated by quantitation of Lts released from human granulocytes stimulated with calcium ionophore, A23187. The method is simpler, less expensive, and more specific for LTD4 than the other methods such as high pressure liquid chromatography and radioimmunoassay and is suitable for routine measurement of either LTD4 specifically or LTC4 plus LTD4 simultaneously in one cell system. PMID- 3035612 TI - Neurotoxicant sensitive esterase. Enzymology and pathophysiology of organophosphorus ester-induced delayed neuropathy. PMID- 3035614 TI - LTB4 mediated hypoxemia in guinea pigs: relationship to pulmonary and cardiovascular pathophysiology. AB - LTB4 is released in the presence of lung injury and may therefore play a role in the pathophysiology of the lung damage. We therefore, administered LTB4 as an I.V. bolus or as an aerosol to guinea pigs and assessed the physiologic response and the lung histology. After 2 ug of I.V. LTB4 airway pressure (AP) rose transiently by 5 +/- 1 mmHg and at five min was back to baseline while PaO2 fell from 96 +/- 5 mmHg to 78 +/- 3 mmHg and remained low at least 45 min. Static compliance (Cstat) was unchanged. Right ventricular systolic pressure (RVSP) and mean aortic pressure (MAP) rose from 9 +/- 1 to 16 +/- 1 mmHg and 43 +/- 4 to 62 +/- 5 mmHg respectively while cardiac index (C.I.) fell from 266 to 208 ml/kg/min but all values were baseline again by 10 min. Aerosolized LTB4 raised AP by 4.6 +/- 0.2 mmHg while PaO2 fell from 90 +/- 7 to 52 +/- 5 mmHg. AP recovered by 20 min but PaO2 remained low at least for 1 hour. MAP, RVSP and CI and Cstat were unaffected. Both I.V. and inhaled LTB4 increased neutrophil infiltrate in the lung although the water aerosol control did too, preventing us from showing a significant effect with LTB4 aerosol. Indomethacin blocked the airway effects and the hypoxemia after I.V. or aerosolized LTB4 but not the neutrophil infiltrate or the rise in RVSP. It actually enhanced (p less than .05) the rise in MAP after I.V. LTB4. Thus cyclooxygenase released products likely mediated the rise in airway pressure and the prolonged fall in PaO2 after LTB4 in guinea pigs but not the pulmonary and systemic vasoconstriction. PMID- 3035615 TI - Desensitization and antagonism of rat polymorphonuclear leukocytes stimulated with PAF acether. AB - The activation of rat polymorphonuclear leukocytes (PMN) with PAF-acether (platelet-activating factor) was blocked by two antagonists: 48740 RP and BN 52021. The release of beta glucuronidase was usually better inhibited than that of lysozyme suggesting that the tested antagonists interfere rather with PAF acether exocytosis of azurophil granules. Inhibition was relatively specific, even though a moderate effect (below 34%) upon PMN activation by the two unrelated agonist n-formyl-Methionyl-Leucyl-Phenylalanine (fMLP) and leukotriene B4 sometimes reached significance. The partial persistence of inhibition to stimulation with PAF-acether after elimination of both inhibitors suggests that they do not act at the PAF-acether receptor only. Furthermore, the observation of cross desensitization between PAF-acether and fMLP may be related to common pathways and/or metabolites, including PAF-acether itself. PMID- 3035616 TI - In vitro action of PG F2 alpha on progesterone and cAMP synthesis in small bovine luteal cells. AB - The action of prostaglandin F2 alpha (PG F2 alpha) on incubated small bovine luteal cells in the presence or in the absence of bovine luteinizing hormone (LH) or dibutyryl cyclic adenosine monophosphate (db cAMP) was investigated. In the absence of LH and db cAMP, PG F2 alpha stimulated progesterone synthesis at concentrations of 10 ng/ml and 100 ng/ml but had no effects at concentrations below 1 ng/ml. PG F2 alpha partially inhibited the LH or db cAMP stimulated progesterone synthesis. This inhibition was maximal for PG F2 alpha concentrations around 100 pg/ml whereas distinctly higher or lower concentrations were without effect. At the concentration of 100 pg/ml, PG F2 alpha partially inhibited the LH induced cAMP accumulation. These results demonstrate an "in vitro" action of PG F2 alpha on bovine luteal cells. They indicate that the luteolytic action of PG F2 alpha in the bovine species could involve, as already suggested for the rat, both an inhibition of the LH induced synthesis of cAMP and an inhibition of the action of cAMP. PMID- 3035617 TI - Evidence for a dual pathway in platelet activating factor-induced aggregation of rat polymorphonuclear leucocytes. AB - The purpose of this study was to determine the role, if any, of Leukotriene B4 (LTB4) in Platelet Activating Factor (PAF)-induced aggregation of rat polymorphonuclear leucocytes (PMNs). Exposure of rat PMNs to 10(-7) M PAF resulted in the release of 4.5 +/- 0.7 ng/10(7) cells of LTB4 measured by radioimmunoassay. However, the maximum aggregation of PMNs achieved by exposure to LTB4 (10(-7)M) was only 50% of that produced by maximally aggregating concentrations of PAF (10(-7)M). 5-Lipoxygenase inhibitors, BW755c and Nafazatrom at concentrations that completely abolished LTB4 synthesis inhibited the aggregation induced by PAF only by 40% and 50% respectively. Furthermore, desensitisation experiments revealed that the aggregatory response of PMNs to PAF was only partially refractory to prior treatment with LTB4 whereas the aggregatory response to LTB4 was completely refractory to prior treatment with PAF. These results suggest that PAF-induced aggregation of rat PMNs is in part mediated by LTB4 and in part directly by an as yet unidentified mechanism. PMID- 3035618 TI - Analogs of leukotriene B4: effects of modification of the hydroxyl groups on leukocyte aggregation and binding to leukocyte leukotriene B4 receptors. AB - The syntheses and agonist and binding activities of 5(S)-hydroxy- 6(Z), 8(E), 10(E), 14(Z)-eicosatetraenoic acid (12-deoxy LTB4), 5(S), 12(S)-dihydroxy-6(Z), 8(E), 10(E), 14(Z)-eicosatetraenoic acid (12-epi LTB4), 12(R)-hydroxy-6(Z), 8(E), 10(E), 14(Z)-eicosatetraenoic acid (5-deoxy LTB4), 5(R), 12(S)-dihydroxy-6(Z), 8(E), 10(E), 14(Z)-eicosatetraenoic acid (5-epi LTB4), 6(Z), 8(E), 10(E), 14(Z) eicosatetraenoic acid (5, 12-deoxy LTB4) are described. These leukotriene B4 analogs were all able to aggregate rat leukocytes and compete with [3H] leukotriene B4 for binding to rat and human leukocyte leukotriene B4 receptors with varying efficacy. The analog in which the 12-hydroxyl group was removed was severely reduced both in agonist action (aggregation) and binding. The epimeric 12-hydroxyl analog demonstrated better agonist and binding properties than the analog without a hydroxyl at this position. In contrast, in the case of the 5 hydroxyl the epimeric hydroxyl analog had greatly reduced agonist and binding activities while the 5-deoxy analog demonstrated potency only several fold less than leukotriene B4 itself. The dideoxy leukotriene B4 analog was more than a thousand fold less active than leukotriene B4 as an agonist and in binding to the leukotriene B4 receptor. These results show that binding to the leukocyte leukotriene B4 receptor requires a hydroxyl group at the 12 position in either stereochemical orientation but that the presence of a hydroxyl at the 5 position is less important. However, the epimeric C5 leukotriene B4 analog clearly interacts unfavourably with the binding site of the leukotriene B4 receptor. PMID- 3035619 TI - The detection of 5-lipoxygenase and cyclo-oxygenase products in sputum of patients with chronic bronchitis and bronchiectasis. AB - Leukotrienes (LTs) and prostanoids (Ps) were detected in sputum of patients with chronic bronchitis and/or bronchiectasis (CB/B) using selective superfusion bioassay and radioimmunoassay (RIA) techniques. Analysis of sputum extracts showed a 4-fold increase in the level of LTB4 compared to the cysteinyl containing LTs (LTC4/LTD4). The measurement of cyclo-oxygenase products (COPs) indicated relatively greater amounts of the vasodilator prostaglandin E2 (PGE2) and prostacyclin (PGI2) compared to the vasoconstrictor prostaglandin F2 alpha (PGF2 alpha) and thromboxane A2 (TxA2) agents (70:30% of total COPs respectively). The presence of eicosanoids (LTs and Ps) in sputum of patients with CB/B suggest that these biologically active substances may act as mediators of bronchoconstriction and inflammation in these diseases. PMID- 3035620 TI - Critical considerations in the development of an assay for sulfidopeptide leukotrienes in plasma. AB - A sensitive and specific assay has been developed for measurement of total sulfidopeptide leukotrienes (LT) in plasma. LTC4 and LTD4 in plasma are converted to LTE4 which is then extracted by C18 Sep-Pak binding and elution. Total LTE4 is resolved by reverse phase high performance liquid chromatography (RP-HPLC) and quantitated by radioimmunoassay (RIA). A [3H]LTE4 internal standard is added to the starting plasma sample to allow overall recovery to be calculated and to define the fractions from RP-HPLC to be assayed for LTE4-like immunoreactivity. The correlation between the measured increase in LTE4 concentration after addition of incremental amounts of LTC4 and LTE4 to plasma was 0.989 and 0.978, respectively, with slopes of 1.05 and 1.11. Addition of 51 pg/ml LTE4 to 5 ml plasma was detectable; the measured increase was 48 +/- 12 pg/ml (mean +/- SE, n = 7). The intra-assay coefficient of variation for 341 pg/ml of added LTC4 was 3.2% (n = 6). Sulfidopeptide leukotrienes could not be detected in blood samples taken from 12 normal volunteers in whom the theoretical detection limit, calculated from the sensitivity of the RIA, the overall recovery of LTE4, and the volume of plasma extracted, was 83 +/- 4 pg LTE4/ml plasma (0.19 +/- 0.01 pmol sulfidopeptide leukotriene/ml plasma; mean +/- SE). PMID- 3035622 TI - [Metastases of breast cancer to the scalp]. PMID- 3035621 TI - Serum relaxin concentrations in women following the administration of 16,16 dimethyl-trans-delta 2-PGE1 methyl ester during early pregnancy. AB - Serum relaxin was estimated in 11 women during termination of first-trimester pregnancy with 16,16-dimethyl-trans-delta 2-PGE1 methyl ester (16 DM-PGE1). Vaginal administration of 16 DM-PGE1 was associated with a significant increase in serum relaxin. PMID- 3035624 TI - A calmodulin antagonist inhibits histamine-stimulated acid production by isolated rat parietal cells. AB - The role of calmodulin in the regulation of histamine-stimulated parietal cell function was studied in isolated rat parietal cells using [14C]aminopyrine uptake as a quantitative index of acid production. In enriched (77-87%) intact parietal cells the calmodulin antagonist naphthalene sulfonamide W 7 dose-dependently inhibited the response to 10(-4) M histamine (IC50: 2 X 10(-6) M). The mechanism of this inhibition was examined further with two other stimuli of H+-production: forskolin which directly activates the parietal cell adenylate cyclase without interacting at the histamine H2-receptor and dbcAMP which mimics the biological action of cAMP without preceding activation of adenylate cyclase. W 7 effectively inhibited the responses to 10(-4) M forskolin (IC50: 6 X 10(-7) M), 10(-3) M dbcAMP (IC50: 10(-6) M) and to 10(-2) M K+ (IC50: 3 X 10(-6) M). The action of W 7 followed non-competitive kinetics since the antagonist reduced the entire range of the concentration-response curves without shifting them rightwards towards higher concentrations of the respective stimulants. The effect of W 7 was reversed by washing the cells. ATP-induced [14C]aminopyrine uptake into digitonin permeabilized oligomycin-inhibited parietal cells reflects H+-production independent of oxidative phosphorylation and was also inhibited by W 7 (IC50: 10( 5) M). Inhibition of K+-stimulated H+/K+-ATPase activity required even higher W 7 concentrations (IC50: 1.4 X 10(-4) M). Our data suggest that calmodulin might be involved in the intracellular mediation of the response to histamine. Between histamine-induced cAMP-generation and the H+-secreting tubulovesicular system W 7 seems to inhibit an intracellular step that finally activates the H+/K+-ATPase. Yet, direct inhibition of the ATPase requires W 7 concentrations of questionable specificity and is unlikely to be the mechanism behind the action of W 7 on the parietal cell response to histamine. PMID- 3035623 TI - [Extrasystemic factors in the pathogenesis and prevention of porphyria cutanea tarda]. PMID- 3035625 TI - Significance of Ca2+, Rb+ fluxes, of cAMP and cGMP for the CCK8-modulated insulin release. AB - In rat pancreatic islets the effects of cholecystokinin-8 (CCK8) on glucose mediated insulin release, 45Ca2+ net uptake, 45Ca2+ efflux, 86Rb+ efflux, cAMP- and cGMP levels were studied. In the presence of a substimulatory glucose concentration (3 mM) CCK8 concentrations of up to 1 microM had no effect on insulin release, but CCK8 at 10 nM potentiated the stimulatory effect of glucose (11.1 mM). 10 nM CCK8 enhanced glucose-stimulated 45Ca2+ net uptake but was ineffective at substimulatory glucose levels. CCK8 had no effect on cAMP and cGMP levels in the presence of 11.1 mM glucose, CCK8 increased 86Rb+ (a measure of K+) in the presence of both 3 and 11.1 mM glucose. This effect was abolished when Ca2+ was omitted from the perifusion medium. CCK8 did not alter glucose (11.1 mM) stimulated 45Ca2+ efflux rate. These data indicate that (1) CCK8 potentiates glucose-stimulated insulin secretion possibly via an effect on Ca2+ uptake, 2) by affecting Ca2+ uptake, CCK8 enhances K+ efflux, and 3) CCK8 does not mediate its effect via cAMP or cGMP. With respect to 86Rb+ efflux the mechanism of CCK8 action appears to be different from that of glucose. When the mechanism of CCK action on islets is compared with that on exocrine pancreas (data from others) there are similarities (importance of Ca2+ uptake and non-importance of cAMP and cGMP). PMID- 3035626 TI - [Hepatocholangiocarcinoma associated with renal clear cell carcinoma]. PMID- 3035627 TI - [Paget's carcinoma of the breast. Clinico-pathological considerations on 85 cases]. PMID- 3035628 TI - [Malignant ovarian germ cell tumors in infancy and adolescence. 14 years' experience]. PMID- 3035629 TI - [Wilms' tumor: histopathology and prognosis]. PMID- 3035630 TI - [Clinical results of dual-energy radiography in thoracic studies]. AB - Dual energy methods permit the reconstruction of either soft tissue or calcium density structures. The value of the technique is demonstrated in 220 patients. Demonstration of the thoracic organs without superimposition of the skeleton improves the recognition of pulmonary lesions. Opacities related to ribs no longer simulate pulmonary lesions. The mediastinal borders are demonstrated more clearly by double spectrum radiography than by any other radiographic method. Soft tissue calcification can be shown with a high degree of sensitivity. Skeletal changes which may be obscured by overlapping soft tissue shadows on conventional radiography can also be diagnosed by this technique. PMID- 3035631 TI - [The thoracic picture in acute traumatic aortic rupture]. AB - We tried to find out the validity of 16 wellknown signs indicating an acute traumatic aortic rupture on plain chest radiographs of 22 patients. Angiographically 11 of all patients had a tear at the aortic isthmus. It turned out that 7 of the 16 signs (widened mediastinum, loss of the aortic knob contour, opacification in the aortopulmonary window, bulging of the vascular pedicle predominantly to the left, left apical cap, depression of the left main stem bronchus and displacement of the right paraspinous interface) are of great diagnostic value. PMID- 3035632 TI - [Pulmonary manifestations of malaria]. AB - We report on the two different types of pulmonary manifestations in acute plasmodium falciparum malaria. The more severe variant shows long standing interstitial pulmonary infiltrates, whereas in the more benign courses only short term pulmonary edemas are visible. PMID- 3035633 TI - [The importance of x-ray cinematography of deglutition for indicating the need for myotomy of the pharyngoesophageal sphincter]. AB - Fourteen patients were examined one to four years after cricopharyngeal myotomy that had been carried out because of dysfunction of the pharyngo-esophageal sphincter. Twelve patients were examined radiologically. Eleven of the 14 patients were clinically improved or cured. In two patients who were not improved, the underlying condition was a polymyositis. The other patients suffered from an idiopathic dysfunction. Because of the small numbers involved, no detailed statistical analysis was carried out. Nevertheless, our results indicate that: Cricopharyngeus myotomy produces marked improvement or cure in patients with idiopathic dysfunction. Weak propulsive peristalsis of the pharyngeal constrictors is a prognostic factor indicating a poor clinical result of surgery. There is little chance of clinical improvement in patients with polymyositis. PMID- 3035634 TI - The Kock continent ileostomy--radiology of dysfunctions. AB - Some patients who have undergone coloproctectomy due to colitis or polyposis with a continent ileostomy constructed ad modum Kock reveal postoperative, secondary abnormalities and dysfunction. The radiological appearance of the pouch in six such patients who complained of incontinence and/or difficulties to catheterize is reported and the findings are correlated with the findings at subsequent surgery and/or endoscopy. It can be concluded that radiology can contribute substantially to the morphodynamic assessment of patients with dysfunction of a Kock ileostomy whereas endoscopy easily depicts mucosal lesions. PMID- 3035635 TI - [Magnetic resonance tomography (MRT) of the adrenals]. AB - Thirty-eight MRT examinations were carried out in 35 patients who, between them, had 19 abnormalities in the adrenal glands. These included ten adenomas, three metastases, two carcinomas, two phaeochromocytomas, and two primary retroperitoneal tumours. MRT provides images of normal and abnormal adrenal glands which are equivalent to those of CT, with the exception that it cannot demonstrate calcification. Using a ratio of signal intensity from the adrenal tumours and the liver on T2-weighted images, it is usually possible to differentiate adenomas from metastases, carcinomas or phaeochromocytomas. The increased specificity of MRT is valuable in the investigation of patients with asymptomatic enlargement of the adrenal glands. PMID- 3035637 TI - [Mammography after breast-preserving therapy of breast cancer]. AB - Follow-up after tumorectomy and intensive irradiation for carcinoma of the breast creates new diagnostic problems. In particular, more experience is necessary about the value of mammography. We have therefore analysed the mammograms of 104 patients intensively followed over one to eight years after primary treatment. Changes in the appearances of the breast due to irradiation are compared with mammographic changes caused by recurrence of tumor. It can be shown that definite structural patterns can be recognised regularly. Optimal diagnosis of recurrencies, however, is only possible by comparing all follow-up mammograms, and additionally the pre-operative films. Close cooperation between the surgeon, radiotherapist and diagnostic radiologist is essential. PMID- 3035636 TI - [Value and indications for high-resolution real-time sonography in nontumor salivary gland diseases]. AB - 308 patients with salivary gland diseases were investigated using real-time sonography. In 142 patients we suspected neoplasms of the salivary glands on the sonograms. In the remaining 166 patients we investigated the following entities: sialolithiasis (74 patients), acute sialadenitis (54 patients), abscess formation (16 patients), chronic sialadenitis or autoimmune disease (53 patients) and cysts of salivary gland (5 patients). The changes in the sonographic morphology of the different diseases will be described. The role of ultrasound in the diagnosis of the different diseases will be discussed and the indications for sonographic investigations are summarised. PMID- 3035638 TI - [Differential sonographic diagnosis of scrotal diseases]. AB - High-resolution scrotal sonography proves valuable in addition to clinical investigation. On the basis of a retrospective analysis of 282 sonographic examinations this method's contributions to the differential diagnosis of diseases of the scrotum are shown. It is usually possible to distinguish testicular from extratesticular tumours; certain signs of inflammation allow the distinction between orchitis and tumour. Sonography is very important for the study of acute painful lesions of the scrotum, which are not palpable, such as rupture following a trauma, severe inflammation and even tumours. PMID- 3035639 TI - [Sonographic detectability of narcotic drug containers in the gastrointestinal tract. Experimental research in the dog]. AB - A "body-pack" is a swallowed plastic or rubber container used by drug smugglers to carry drugs. We report our experience in diagnoses of such packs in vivo. We simulated the pack by stuffing a fingerstall of a latex handglove with a mixture of 10% barium powder and flour to render it radiopaque. Eight serial examinations were made. In each series, an adult Alsatian dog was fed upto 10 such "body packs". The passage of these packs through the gastrointestinal tract was followed with ultrasound examinations made at 2, 24, and 48 hours postprandially. Immediately after each sonographic localisation of "body-pack" was made, an x-ray examination of the abdomen in this region was also carried out. Ultrasound correctly determined the position of the "body-pack" in 20 out of 24 examinations as compared to x-ray results. In 4 cases ultrasound could not confirm the location of the "pack". PMID- 3035640 TI - Synovial chondromatosis of the ankle. AB - Synovial chondromatosis is a rare disorder presenting a typical radiological appearance. We reviewed three patients with this disease in the ankle. Two of these patients were reexamined after 7 and 27 years, respectively. Although both had several calcified juxta-articular bodies in the ankle none of them had any clinical symptoms. This indicates that surgical intervention should be performed only if mechanical signs and symptoms are present. PMID- 3035641 TI - [Pedicle agenesis in the cervical spine]. AB - Congenital absence of a cervical pedicle is rare, and because of the associated enlargement of the intervertebral foramen, may be confused with a "dumbbell" tumour. A recent observation of this anomaly has prompted this report. Based on the findings of 29 reported cases in the literature, the characteristic radiological features of the abnormality are described and the differential diagnosis as well as the clinical implications of this disorder are discussed. PMID- 3035642 TI - [Uptake behavior of bone metastases of hypernephroma in the 99m Tc-MDP bone scintigram. A comparison with x-ray findings]. AB - Ninety-one radiologically confirmed osteolytic metastases in 30 patients with hypernephromas were studied with regard to their uptake of 99mTc-MDP and this was compared with the radiological findings. In 16% of the radiologically proven metastases, there was no correlation with their isotope uptake. Compared with other bone metastases whose isotope uptake has been studied and described in the literature, there appears to be a higher proportion of hypernephroma secondaries that do not show uptake of 99mTc-MDP. PMID- 3035643 TI - [Long-term follow-up of vena cava filters: real-time sonography and the native x ray image]. AB - In 16 patients with vena cava filters (12 Gunther filters, 4 Kimray-Greenfield filters) a follow-up study was performed by ultrasound and abdominal plain film. Ultrasound allows to identify type and location of the filter as well as the entrapment of emboli. Abdominal radiography is useful for exact evaluation of changes in filter position. Combination of sonography and plain films is considered to be most suitable for follow-up studies of caval filters. PMID- 3035644 TI - Low-dose streptokinase in the treatment of arterial graft occlusions. AB - During the past 24 months, 19 infrainguinal bypass graft occlusions occurring in 17 patients were treated with local infusions of streptokinase. The treated grafts included three venous femorocrural grafts and four venous and 13 Goretex femoropopliteal grafts. Infusion managed to restore flow completely in 10 grafts. Partial clot lysis occurred in three and no lysis in six grafts. All successful recanalisations occurred within 120 hours of the initiation of thrombolytic therapy. Despite three major complications and the development of two minor haematomas, with an overall success rate of 55% our results support the use of streptokinase in the management of occluded bypass grafts. PMID- 3035645 TI - [Computed tomographic research on the viability of a free fat pedicle flap in the area of a laminectomy]. AB - Nineteen patients were examined clinically and by CT three to five years after partial or total hemilaminectomy using a fat pedicle as cover. The results were compared with a control group of fourteen patients without fat pedicles. With CT it is possible to check the fat pedicles directly. Survival of the fat pedicle without peridural scarring was present in 68%. It was not possible to demonstrate any correlation between the extent of scarring and clinical abnormalities. PMID- 3035646 TI - [MR and CT patterns of neurocysticercosis]. AB - MRI and CT manifestations were studied in five cases of neurocysticercosis. As demonstrated by long-term follow-ups the disease usually causes multiple lesions the morphology of which depends on the life cycle of the parasite. Tissue lesions consist of three main types: 1) vital cysticerci, 2) inflammatory parenchymatous reactions following degenerating cysts and 3) calcified granulomas. MRI provides all information that is given by CT except for small calcifications which are usually missed. Morphological details of vital cysticerci like cyst wall and scolex are better outlined by MRI. When i.v. contrast medium is applied, it leads to nodular or annular enhancement of inflamed tissue. The sensitivity of MRI towards edema caused by parasite exceeds that of CT by several weeks. CT and MRI are complementary methods providing at the present time the highest degree of specificity in diagnosing neurocysticercosis. PMID- 3035648 TI - [RARE-MR myelography in routine clinical practice. Experience with 175 cases]. AB - In 1984 Henning, Nauerth, Friedburg and Ratzel described a new MRI data acquisition technique called RARE (Rapid Acquisition with Relaxation Enhancement). Several clinical applications have been published. A modification developed in the meantime, called "RARE hydrography", yields T2-weighted aqueous fluid specific images within a few seconds. As a nontomographic rapid approach to MR-myelography ("RARE-myelography") this technique is routinely applied at the beginning of all examinations of the spine. On the basis of 175 cases current potentials and limitations are discussed. Future applications of 3D-RARE myelography are mentioned. PMID- 3035647 TI - [Magnetic resonance imaging in craniopharyngiomas. The differential diagnosis of cystic intra- and suprasellar space-occupying lesions]. AB - Seven patients with confirmed craniopharyngiomas were examined via magnetic resonance imaging. The various tumour components (whether solid, cystic or calcified) were evaluated separately. Three types of cysts were observed that can be distinguished by their signal intensity in various pulse sequences. This could be explained largely by an analysis of the T1 and T2 image contrast. The reason for the various types of cysts is determined by their cholesterol content. MR imaging is not as useful as CT for demonstrating the solid and calcified portions of the tumour and this reduces its value in diagnosing the type of tumour. Against that, it is superior with regard to separating the tumour from its neighbouring structures. The differential diagnosis, which must be considered in using MR, is summarised in a table at the end of the paper. PMID- 3035650 TI - [Myxoma of the maxilla. Demonstration of computerized tomography findings apropos of a case]. PMID- 3035649 TI - [Initial clinical results of tissue characterization by T1, T2 and proton density in nuclear magnetic resonance tomography]. AB - The NMR parameters (proton density, relaxation times T1 and T2) have been assessed by Carr-Purcell-Meiboom-Gill (CPMG) spin echo sequences. A computer assisted analysis of the data of 21 patients with cerebral tumours allowed a classification of tumour tissue in different tumours. The use of quantitative procedures for tissue characterisation allows the differentiation of benign and malignant brain tissue by characteristic colour coding demonstrating morphological details like tumour, edema and necrosis as well as indicating the histological types of the tumours of the central nervous system. PMID- 3035652 TI - [Toxocariasis (visceral larva migrans) of the liver. The computed tomographic aspects]. PMID- 3035653 TI - Metastatic abscesses resulting from staphylococcal infection of the hand: CT appearance. PMID- 3035651 TI - [Mediastinal metastasis in osteogenic sarcoma. X-ray and nuclear medicine follow up]. PMID- 3035654 TI - [Diagnosis of urine leak after kidney transplantation]. PMID- 3035655 TI - [Demonstration of the lateral thoracic vein in intrathoracic inlet obstruction by magnetic resonance]. PMID- 3035656 TI - [Tsetse control (Glossina: Diptera, Muscidae) using screens and traps (static methods): evaluation of 2 years' control in Sirasso in the northern Ivory Coast]. PMID- 3035657 TI - [Adenoid cystic carcinoma of the trachea. Treatment by combined laser therapy and surgery]. AB - In the group of adenomas of the trachea and bronchi, cystic adenocarcinomas or cylindromas are malignant tumours, whose outcome is much slower than those of the usual type of cancer. An unusual case is reported in which treated was firstly carried out with laser therapy and then surgery. PMID- 3035658 TI - Calorigenic action of circulating epinephrine during pentobarbital, halothane and morphine anesthesia in dogs. AB - The calorigenic action of circulating epinephrine was analyzed quantitatively in the dog during pentobarbital, halothane and morphine anesthesia. The whole body oxygen consumption (VO2) was measured by an on-line analysis of breath-by-breath respiratory gas exchange. Epinephrine was infused at various doses and the plasma concentrations of epinephrine were measured by semiautomated fluorimetric analysis. During pentobarbital anesthesia, the basal VO2 was 5.26 ml/kg/min and was increased by epinephrine in a dose dependent manner at plasma concentrations between 3.9 ng/ml (VO2 = 5.68 ml/kg/min) and 36.5 ng/ml (VO2 = 6.47 ml/kg/min). Epinephrine exerted a similar effect during halothane anesthesia, but it failed to affect VO2 during morphine anesthesia. Morphine also inhibited dibutyryl cyclic AMP-induced increase in VO2, suggesting that morphine inhibits the calorigenic action of epinephrine at the intracellular site distal to beta receptors. PMID- 3035659 TI - Comparison of digitoxin and its major glucuronidated metabolite: inhibition of Na K-ATPase and reactivity with the radioimmunoassay. AB - The metabolism of digitoxin is known to occur through several major pathways including oxidation and glucuronidation. Several studies have compared the behavior of the various unconjugated metabolites with respect to reactivity toward Na-K-ATPase and toward the radioimmunoassay (RIA). Other studies with conjugated products synthesized chemically have also been performed. However, the activity of the conjugated products produced in vivo has not been reported and these metabolites are generally assumed to be inactive. In the present studies we have prepared and purified the glucuronide conjugate of digitoxigenin monodigitoxoside formed by rat liver microsomes and determined the activity of this metabolite as measured by inhibition of Na-K-ATPase and by the RIA. The concentration of the conjugated product needed to cause a fifty per cent inhibition of Na-K-ATPase was 5.40 microM compared to 0.68 for digitoxin and 0.08 for digitoxigenin monodigitoxoside. However, the conjugated product had a two fold greater affinity for the antibody in the RIA procedure than did either digitoxin or digitoxigenin monodigitoxoside. Although these data cannot be extrapolated to the effects of these drugs on cardiac contractility, the results suggest that the contribution of the glucuronide conjugate to the therapeutic or toxic effect of digitoxin may be minimal. This metabolite may, however, lead to inaccurate estimation of blood levels by the RIA. PMID- 3035660 TI - Efficacy of a bivalent vaccine against Marek's disease. AB - A bivalent vaccine was prepared by combining inactivated Marek's disease virus and turkey herpesvirus. The efficacy of this vaccine, compared to turkey herpesvirus and inactivated Marek's disease virus separately, was studied in unsexed White Leghorn chicks which were vaccinated at one day old and then challenged at 21 days old with fowl blood infected with virulent Marek's disease virus. The bivalent vaccine appreciably delayed mortality resulting from Marek's disease and elicited the highest protective efficacy as judged on the basis of Marek's disease-specific mortality and percentage occurrence of lesions. The occurrence, extent and severity of gross lymphomas and microscopic lymphoproliferative lesions in various organs of the bivalent vaccinated birds were less than in the other challenged groups. In addition, the level of viraemia remained consistently and significantly lower in the bivalent vaccinated birds. PMID- 3035661 TI - Duck hepatitis virus: adaptation of a plaque assay to determine 50 per cent end points with duck sera. AB - An assay to determine 50 per cent neutralisation end points of duck anti-duck hepatitis virus sera was developed using a plaque assay to quantify residual virus. The optimal conditions were determined. Ducklings produce a serological response within four days of vaccination and the response reaches a maximum within nine days. PMID- 3035662 TI - Local humoral and cellular responses in Aujeszky's disease virus infection in pigs. AB - Mucosal and tracheal washings from pigs vaccinated parenterally and intranasally with Aujeszky's disease virus were tested for specific anti-Aujeszky's disease virus responses. Antibody tests included complement dependent antibody lysis, antibody dependent cellular cytotoxicity, virus neutralisation, and anti Aujeszky's disease virus IgA and IgG levels. There was no correlation between the levels of these antibodies and protection from subsequent challenge. Direct lymphocyte cytotoxicity against cells infected with Aujeszky's disease virus was found in lymph nodes draining the tonsillar area. PMID- 3035663 TI - Effect of bovine virus diarrhoea-mucosal disease virus infection on salmonella infection in calves. AB - The possibility that bovine virus diarrhoea virus (BVDV) infection could aggravate concurrent salmonella infections was investigated in two series of experiments with Salmonella dublin and S typhimurium. In both series of experiments the clinical signs tended to be more severe in dual infections than in those with salmonella alone. Faecal excretion of S dublin was similar in both single and dual infected calves. A protracted bacteraemia occurred only in two dual infected calves, one of which died from suppurative meningitis. In the case of S typhimurium, body temperatures, passage of abnormal faeces and isolation rates of salmonella from faeces were significantly increased. The results suggest that BVDV infection may indeed exacerbate the effects of salmonella infection and the aetiological relationship of the two agents should be considered during outbreaks of bovine salmonellosis. PMID- 3035664 TI - Incidence of rotavirus in beef herds in Argentina. AB - Faecal samples were collected from 177 diarrhoeic and 40 healthy calves from 19 farms in Buenos Aires State, Argentina during 1984 and 1985. Samples were examined for rotavirus by enzyme-linked immunosorbent assay (ELISA) and polyacrylamide gel electrophoresis (PAGE) of their genomic RNA segments. Rotavirus was found in 95 samples of diarrhoeic calves (53 per cent) and in three healthy calves (7 per cent). All positive samples were tentatively classified as group A on the basis of electropherotype and ELISA test reactivity and exhibited 18 different genomic electropherotypic patterns. PMID- 3035665 TI - Proceedings of a conference on resuscitative thoracotomy. New York City, February 21, 1986. PMID- 3035666 TI - Resuscitative thoracotomy: introduction. PMID- 3035667 TI - Open versus closed chest cardiac massage in non-traumatic cardiac arrest. AB - Since the rediscovery and popularization of closed-chest cardiopulmonary resuscitation (CPR) in the early 1960s, this technique has largely replaced open chest cardiac massage. However, in the ensuing quarter of a century, a large amount of data has been accumulated that seems to indicate that open-chest CPR is the physiologically superior method of cardiopulmonary resuscitation. This paper reviews that data comparing these two CPR modalities. The first part discusses data obtained from animal studies, while the second considers the limited amount of information that has been obtained from human investigations. The authors concludes that a randomized clinical trial of open-chest and closed-chest CPR is needed to fully evaluate the efficacy of these two resuscitative techniques as well as to define the most appropriate circumstances for the use of internal cardiac massage. PMID- 3035668 TI - Emergency department thoracotomy for trauma: a collective review. AB - A decade of experience with resuscitative thoracotomy for the trauma victim in extremis has been gained since the pioneering efforts of Mattox and his associates in 1974. It appears, from a review of the various reports from different trauma centers, that there is an emergence of a consensus as to the best indications for the procedure. It is generally agreed upon that ERT is fruitless in the patient with severe head trauma or when vital signs were absent at the scene of the injury. In the absence of penetrating thoracic injuries ERT yields a very poor survival in patients without vital signs on admission to the emergency center. It is widely accepted that the best results for ERT are in patients with cardiac tamponade. The prognosis is hopeless in patients without vital signs after sustaining blunt trauma. PMID- 3035669 TI - Hemodynamic mechanisms in CPR: a theoretical rationale for resuscitative thoracotomy in non-traumatic cardiac arrest. AB - Experimental work over the past decade has revealed three distinct mechanisms for generating artificial circulation during cardiac arrest and resuscitation. To isolate these mechanisms and study them in pure form, and in particular to characterize circulation during open vs. closed chest cardiopulmonary resuscitation (CPR), we developed an electrical model of the human circulatory system. Heart and blood vessels were modeled as resistive-capacitive networks, pressures in the chest, abdomen, and vascular compartments as voltages, blood flow as electric current, blood inertia as inductance, and the cardiac and venous valves as diodes. External pressurization of thoracic and abdominal vessels, as would occur in CPR, was simulated by application of half-sinusoidal voltage pulses. Simulations included two modes of creating artificial circulation: the cardiac pump mechanism, in which the atria and ventricles of the model were pressurized simultaneously, as occurs during open chest cardiac massage, and the thoracic pump mechanism, in which all intrathoracic elements of the model were pressurized simultaneously, as is likely to occur in closed chest CPR. The two mechanisms were compared for the same peak applied pressure (80 mmHg). Pure cardiac pump CPR generated near normal systemic perfusion pressures throughout the compression cycle. Pure thoracic pump CPR generated much lower systemic perfusion pressure only during the diastolic phase of the compression cycle. Simulation of cardiac compression at rates from 40 to 100/min produced total flows of 2500-3300, myocardial flows of 150-250 and cranial flows of 600-800 ml/min, depending on the compression rate. In contrast, thoracic pump CPR produced a total flow of approx. 1200, myocardial flow of 70, and cranial flow of 450 ml/min, independently of the compression rate. Direct cardiac compression is an inherently superior hemodynamic mechanism, because it can generate greater perfusion pressure throughout the compression cycle. If one presumes that improved blood flow during CPR is the key to more successful resuscitation, then it is reasonable to conclude that direct heart massage is the most effective available way to achieve this end. PMID- 3035670 TI - Open-chest cardiac massage after closed-chest compression in a canine model: when to intervene. AB - Open-chest cardiac massage appears beneficial in improving hemodynamics during resuscitation efforts and in improving resuscitation success. The time between cardiac arrest and the initiation of open-chest cardiac massage is crucial. It would appear that if initiation of open chest cardiac massage is delayed for more than 20 min from the onset of cardiac arrest, little or no successful outcome can be expected. Further techniques for assessing the adequacy of closed-chest compression CPR are needed. Such techniques would allow early identification of ineffective resuscitation efforts and provide the opportunity for early change to other, presumably more effective, techniques. PMID- 3035671 TI - Open chest cardiac massage: an overview. AB - Open-chest cardiopulmonary resuscitation has been shown to produce better blood flow in man than closed-chest massage. It therefore should be taught as part of the protocol for all hospital CPR teams. PMID- 3035672 TI - [Changes in beta-adrenergic receptor density in congestive heart failure and myocardial hypertrophy]. PMID- 3035673 TI - [Effects of PGI2 analogue (CS-570) in patients with acute myocardial infarction: dose response effects on cardiovascular hemodynamics, plasma prostanoids, catecholamines and cyclic nucleotides]. PMID- 3035676 TI - Antiviral activity of naphthoquinones. I. Lapachol derivatives against enteroviruses. PMID- 3035678 TI - [Cell transformation with the plasmid pBR-SV40: isolation and characterization]. PMID- 3035674 TI - Relation between clinical stage of sarcoidosis and serum values of angiotensin converting enzyme and beta2-microglobulin. AB - We measured angiotensin converting enzyme (ACE) and beta2-microglobulin (beta 2M) in the serum of 107 patients with sarcoidosis in different clinical stages and in healthy subjects. In patients with acute sarcoidosis and erythema nodosum (EN) we found increased beta 2M values but low ACE activity. Patients with newly-detected sarcoidosis but without EN had increased ACE activity; however, their beta 2M levels were normal or only slightly elevated. Patients with chronic active sarcoidosis had high ACE activity; however, their beta 2M-levels were normal in a stable phase, but significantly increased during clinical relapses. Patients with resolved sarcoidosis had normal ACE and beta 2M values. The results indicate that serum beta 2M concentrations are increased in connection with new granuloma formation-i.e. lymphocytic activation-whereas the serum ACE activity increases later on when active granulomas have been formed. In sarcoidosis, simultaneous determinations of serum ACE and beta 2M may be useful for describing the stage of the disease. The correlation between ACE and beta 2M will also be different, depending on the composition of the clinical series of sarcoidosis patients. PMID- 3035675 TI - Characterization of peripheral blood T lymphocyte subsets in pulmonary sarcoidosis. AB - The peripheral blood of sarcoidosis patients has been shown to have abnormalities in the T Lymphocyte subsets. There is a consistent reduction in the helper/suppressor T Lymphocyte ratio caused by an increase in suppressor cell percentage as determined by monoclonal antibodies. These findings suggest that the peripheral blood T Lymphocyte subsets may be a useful adjunct for determining the underlying immunological cellular dysfunction in sarcoidosis patients. PMID- 3035677 TI - [Application of the passive hemagglutination test to the study of equine rhinopneumonitis. II. Serologic study in horses]. PMID- 3035679 TI - Influence on blood pressure of oestrogen substitution therapy in the menopause. AB - In a Swedish health centre an analysis was made of the effect on blood pressure of oestrogen substitution therapy in 33 normotensive women in the menopause. Comparisons were made with a control group of 33 normotensive women of the same ages but left without oestrogen therapy. Following three years of therapy with oestrogen alone or in combination with progesterone, there were no significant changes in blood pressure. This was true also when a subgroup consisting of 11 women treated with equinous oestrogens was analysed separately. These results could therefore indicate that chronic treatment with oestrogens in postmenopausal women probably has small effect on the general blood pressure level and that the risk of inducing arterial hypertension must be low. PMID- 3035680 TI - Effect of mechanical stimulus spread across glabrous skin of raccoon and squirrel monkey hand on tactile primary afferent fiber discharge. AB - The role of spread of skin deformation in activating cutaneous mechanoreceptors at a distance from their threshold receptive fields (RFs) was examined in glabrous skin of the North American raccoon and the squirrel monkey. One feedback controlled mechanical stimulus probe was used to indent the skin to a controlled depth at a constant velocity, at varying distances from a second probe, which was used to monitor vertical displacement depth and velocity at this distant site. In many instances, the monitor probe was positioned over the RF of a cutaneous mechanoreceptor, and single-unit action potentials were simultaneously recorded from individual fibers of the median or ulnar nerve. With distance from the site of stimulation, there was a systematic, monotonic decline in indentation depth and velocity; velocity fell off with distance more rapidly than depth. The degree of diminution with distance varied with the size, shape, and curvature of the digital or palm pad stimulated. Spread of indentation was more restricted on digital than on palm pads, and was more restricted across monkey skin than across raccoon skin. Spread was less with higher-velocity than with lower-velocity indentations, but was seemingly unaffected by indentation depth. As expected from the findings noted above, the number of spikes discharged by slowly adapting mechanoreceptive afferent fibers declined more rapidly with distance between stimulus site and RF for digital than for palmar RFs, in squirrel monkey than in raccoon skin, and with higher-velocity than with lower-velocity stimuli. Furthermore, the number of spikes occurring during either ramp or early static indentation phases of stimulation dropped to zero more rapidly with distance than did either vertical indentation depth or velocity. Decreases with distance in both indentation depth and velocity acted to restrict the size of suprathreshold RFs. For most units, horizontal components of mechanical stimulation subtracted from the effects of vertical components. It is suggested, on the basis of this and other studies, that many neural and perceptual phenomena usually attributed to central mechanisms of afferent inhibition may be attributable, at least in part, to mechanical properties of the skin. In addition, the present data suggest that regional variations in the two-point limen may be associated with variations in spread of mechanical deformation. The conclusion that glabrous skin and subjacent soft tissues act as a low-pass filter system provides a mechanical basis for the relative efficacy of high-frequency vibratory stimuli in tactile pattern perception.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3035682 TI - Surveillance in relatives of patients with adenomatous polyposis. AB - Registry ascertainment of kindreds with adenomatous polyposis (AP) reduces the incidence of colorectal carcinoma by medical monitoring for offspring and siblings of affected patients. Due to its pleiomorphic genotype and a 50% risk for AP in each patient's progeny, early screening is mandatory. Flexible sigmoidoscopy, averaging from puberty to age 48, annually or every two years, is one diagnostic technique that does not result in excess patient discomfort or cost. Timely examination is facilitated when eye anomalies, osteomas, or epidermoid cysts are present in childhood as potential clinical markers for AP. Denial, misperceptions, and fear may impede surveillance in the absence of education by the registry health care team. PMID- 3035681 TI - [The many faces of breast cancer]. PMID- 3035683 TI - Causes of death in patients with familial polyposis coli (FPC). AB - Autopsy reports from patients with familial polyposis coli were obtained from the Roswell Park Memorial Institute Registry as well as clinical data on patients with familial polyposis coli with regard to their causes of death. Although the latter group did not undergo autopsy they were the subject of a close follow-up. From this data it is clear that diagnosis at an early age may not diminish the chances of the patient with familial polyposis to develop cancer. Removal of the colonic mucosa does not provide absolute assurance that patients with this condition will be cured, as these patients remain at considerable risk for developing other benign and malignant lesions of the gastrointestinal tract and perhaps also of other organs. Close follow-up of FPC patients who have had surgery for colorectal polyps or cancer is necessary. Other proliferative abnormalities of FPC patients include an abnormal fibroblastic response, tendency to develop adhesions, and the propensity for developing desmoids. Desmoids may follow a slow but unrelenting course with a lethal outcome. Autopsy findings support the view that there should be no distinction between familial polyposis coli and Gardner's syndrome. PMID- 3035684 TI - Activities of the Leeds Castle Polyposis Group. AB - In June 1985 an international meeting took place at Leeds Castle, England, to discuss some of the problems facing those caring for polyposis patients and conducting research in the field. From this grew the Leeds Castle Polyposis Group, comprising workers from 19 centers around the world; they agreed to cooperate initially in the collation of retrospective data with the hope that this would lead to prospective studies including comparative therapy trials. So far four retrospective pro formas have circulated. The data from these and plans for further projects will be discussed at the next meeting of the group in Washington in April 1987. PMID- 3035685 TI - Genetic markers of colonic neoplasia. AB - Adenomas and polyposis syndromes can be viewed as stages preceding the development of colonic cancer. Genetic markers of these stages have potential value in diagnosis and in investigation of biological mechanisms and consequences of cancer-predisposing genes. No reliable markers for premalignant states are yet available, but unambiguous gene markers for familial polyposis will probably be identified in the near future. They will clarify relationships among the hereditary colon cancer syndromes and possibly help to define the sequence of malignant transformation in sporadic cancers as well. PMID- 3035686 TI - Biochemical markers in patients with familial colonic neoplasia. AB - Many potential markers, including biochemical markers, have been studied in an attempt to identify the presence of early colorectal neoplasia or risk of neoplasia, particularly in those families with hereditary colonic neoplasia syndromes. Unfortunately, most of these markers are useless in screening or diagnosis. Nevertheless, such markers as carcinoembryonic antigen (CEA) and CA 19 9 may have a role in pretherapeutic and posttherapeutic monitoring of disease or recurrence. The newer tumor markers, including the carbohydrate markers, ornithine decarboxylase (ODC) and the polyamines, are of great interest as potential tumor markers; ODC and the polyamines may also have a future potential as therapeutic targets. However, further studies are needed to determine their true sensitivity and specificity in hereditary colonic neoplasia syndromes, as well as in patients without genetic syndromes who are at risk for colorectal cancer. PMID- 3035687 TI - Chromosomal abnormalities in patients with familial polyposis and colorectal cancer. AB - A search for chromosomal (cytogenetic, karyotypic) changes with specificity for familial polyposis coli (FPC) and related conditions has not been successful. The cytogenetic anomalies described to date (chromosomal instability, tetraploidy, structural anomalies) are not only nonspecific and inconsistent but may also be seen in other congenital conditions predisposing to cancer of various organs. However, this does not mean that some (eg, tetraploidy) of the changes cannot be used as indices in the study and evaluation of affected individuals and their family members. Future application of DNA probes may be more successful in establishing genetic anomalies unique to FPC and related disorders. PMID- 3035689 TI - Familial polyposis coli. PMID- 3035688 TI - Counseling families with hereditary gastrointestinal polyposis syndromes. AB - Counseling for families with one of the hereditary polyposis and/or colon cancer syndromes can be offered by a number of different professional persons depending upon the emotional needs of the counselee. It is sometimes difficult to persuade at-risk persons in polyposis families to institute a medical surveillance plan with their physicians because of their reactions to knowledge (or lack of it) of the family diagnosis. Counseling may reveal both emotional and financial problems as deterrents to needed medical planning. Support organizations and explanatory literature are helpful in allaying fears and promoting compliance. PMID- 3035690 TI - Historical developments in familial polyposis coli. AB - The emergence of familial polyposis coli from the general group of conditions with multiple polyps of the large intestine into a well-defined separate entity started when histopathology became a science. The recognition of its two main features--inheritance as a Mendelian dominant characteristic and its high incidence of associated colorectal cancer--greatly helped in establishing a policy of treatment designed to prevent cancer. The removal of all or most of the large intestine before cancer had supervened has had considerable success in reducing the cancer incidence. However, the more recent awareness that the adenomas can frequently be found also in the upper gastrointestinal tract has introduced new problems, the solution of which is currently a matter of investigation. PMID- 3035691 TI - Patterns of inheritance of colonic polyps. AB - While the inheritance pattern of familial polyposis coli is established as an autosomal dominant pattern, the expression of the various extracolonic manifestations associated with the neoplastic polyposis is less well understood. The discrete polyp cancer syndrome may not be recognized unless both polyps and colon cancer are considered in the inheritance pattern. The hamartomatous polyps follow a Mendelian-dominant inheritance pattern for the Peutz-Jeghers syndrome, while the inheritance pattern for the juvenile polyposis syndromes is less clear. Cowden's disease appears in a Mendelian dominant pattern, but the occurrence of colonic polyps is less well documented. The ganglioneuromas follow a mendelian dominant inheritance pattern, while the relationship and occurrence of colonic polyps in association with Torre's syndrome is uncertain. The Mendelian dominant inheritance pattern for the cancer family syndrome is documented; the role of colon polyps in this syndrome is less well understood. A further understanding of the inheritance patterns of these various colon polyps will lead to more understanding of the basic disease and help in prevention and early detection for treatment and cure. PMID- 3035692 TI - Dental and bone abnormalities in patients with familial polyposis coli. AB - Dental and bone abnormalities of the maxilla and mandible are present in approximately 80% of patients with familial polyposis coli. The dental abnormalities include impacted teeth (other than third molars), supernumerary teeth, congenitally missing teeth, fused roots of first and second molars, and unusually long and tapered roots of posterior teeth. The bone lesions consist mostly of osteomas, either isolated or in clusters, in the maxilla and mandible or of exostoses with lateral and/or lingual extensions. Since dental and bone abnormalities are already present early in life there is a strong suggestion that they may be used as diagnostic features in the recognition of familial polyposis coli. PMID- 3035693 TI - Incidence of associated diseases in familial polyposis coli. AB - In this review we present diseases associated with hereditary polyposis syndromes. In addition to the known extracolonic manifestations of the so-called Gardner's syndrome, other important diseases are carcinomas of the thyroid and of the small bowel and bile duct adenoma. The incidence of these diseases is discussed. PMID- 3035694 TI - Extracolonic manifestations of familial polyposis coli. AB - The original concept of familial polyposis coli (FPC) as only a genetically determined premalignant disease of the colon changed in the 1950's, with the description of Gardner's syndrome. The extracolonic manifestations of osteoma and epidermoid cyst have since been shown to be only a small part of the spectrum of both benign and malignant extracolonic manifestations of the disease. The modern concept of this condition is that FPC is a genetically determined generalized growth disorder that gives rise to tumors in various parts of the body. PMID- 3035695 TI - Enteric continence and the ileal pouch-anal procedure. AB - The ileal pouch-anal procedure maintains enteric continence by preserving critical physiological mechanisms. The reservoir, although possessing somewhat different physiologic characteristics than the rectum, provides sufficient storage capacity to enable the patient to resume most daily activities. The intact anorectal angle continues to perform its pivotal role in the maintenance of gross fecal continence, while the functioning anal sphincter muscles fine-tune the system. The success of the procedure has shown that certain widely held "principles" of continence may be less important than previously believed. These include the requirement for a long rectal stump, the preservation of the rectosphincteric inhibitory reflex, and the maintenance of sensation to the upper half of the anal canal. Yet, results are not perfect, and it is possible that while these mechanisms are not required for gross fecal continence, their transgression results in a subtle reduction in function from that of the normal state. When one considers, however, that the only other curative procedure for patients with familial polyposis is total proctocolectomy with permanent ileostomy, the ileal pouch-anal operation represents an important advance. The disease is removed, yet reasonably effective control of enteric continence is restored such that quality of life appears to be enhanced when compared to that achieved after proctocolectomy and Brook ileostomy. PMID- 3035696 TI - Surgical alternatives in the treatment of polyposis coli. AB - The history of familial polyposis coli and its various surgical treatment alternatives are presented. Each form of treatment has advantages and disadvantages. The menu of alternatives allows the surgeon the freedom to choose the best option for each patient. PMID- 3035697 TI - Molecular mechanisms for transposition of drug-resistance genes and other movable genetic elements. AB - Transposition is proposed to be responsible for the rapid evolution of multiply drug-resistant bacterial strains. Transposons, which carry the genes encoding drug resistance, are linear pieces of DNA that range in size from 2.5 to 23 kilobase pairs and always contain at their ends nucleotide sequences repeated in inverse order. In some transposons the terminal inverted repeat sequences are capable of independent movement and are called insertion sequences. Transposons carry a gene that encodes transposase(s), the enzyme(s) responsible for recombination of the transposon into another DNA molecule. Studies on transposable genetic elements in bacteria have not only given insight into the spread of antibiotic resistance but also into the process of DNA movement. PMID- 3035699 TI - [Effect of natural low- and high-fiber diets on beta-carotene conversion to vitamin A]. PMID- 3035698 TI - [Cytomegalovirus infection caused by blood transfusion]. PMID- 3035700 TI - [Opioid peptides and their receptors]. PMID- 3035701 TI - Interferon level in human normal sera. AB - Interferon (IFN) level in the sera of normal humans was determined by their protective effect on a KB cell culture against the cytopathic effect (CPE) induced by vesicular stomatitis virus. The technique revealed all the three known types of IFN. In the investigated population 15 of 37 sera (40%) were found to protect against the CPE at 1/10 dilution, 11 of 37 (30%) still protect at 1/20 dilution, and 7 of 37 sera (19%) still protect at a dilution of 1/40. No serum was found to have detectable IFN levels at 1/80 dilution. We can thus consider that the normal serum level ranges up to a 1/20 dilution i.e., up to 20 IU/ml. The above titers are slightly higher than the normal IFN level reported by other investigators in other human populations, although no clear reasons can be given to account for the differences. PMID- 3035702 TI - [Critical study of radiculomedullary and neuromuscular complications of ankylosing spondylitis]. AB - Medullo-radicular and neuro-muscular involvements of ankylosing spondylarthritis, often reported in an analytic fashion in the literature, deserve to be the subject of a critical study. Various neurological manifestations secondary to exceptional atlo-occipital and sometimes axis-atlas subluxations and medullary lesions as well as syndromes of the cauda equina. The medullary lesions have an epidural origin (3 cases in the literature, 2 cases from the authors) or are secondary to a spondylodiscitis (4 cases in the literature) or secondary to both (1 case reported by the authors). As for syndromes of the cauda equina the authors report 3 cases to be added to the 55 published previously. It concerns always old spondylarthritis. The lesions combine posterior diverticula and lesions of the lamina. The treatment is usually ineffective. A special case is represented by forms with trophic disorders. More debatable are the radicular lesions, which, except for intercostal pain, should be linked to local pain. Electromyographic abnormalities are of no significance. Alterations of the paravertebral muscles viewed on the scanner X have, for now, an uncertain significance. Finally, various associations, without significance such as multiple sclerosis, diffuse muscular lesions and the classic spondylotic pseudo tabes, should be rejected. PMID- 3035703 TI - Beta-adrenergic receptors of human lymphocytes in physically active and immobilized subjects: characterization by a polyethylene glycol precipitation assay. AB - To investigate the role of physical conditioning in the regulation of cellular beta-adrenergic responsiveness in man, a simple and accurate method for the determination of lymphocytic beta-adrenergic receptors was developed and subsequently utilized for the characterization of these receptors in subjects with varying degrees of physical fitness. This method combines the advantages of the use of [125I]iodocyanopindolol as the radioligand and polyethylene glycol (PEG) precipitation for the separation of bound and unbound radioligand. The assay is quick and easy to perform and allows several samples to be processed simultaneously. The concentration of beta-adrenergic receptors in lymphocytic membrane particulates in 16 healthy subjects was 58 +/- 4 fmol/mg protein (mean +/- SEM) and the Kd of the interaction between [125I]iodocyanopindolol and the receptor was 4 X 10(-11) mol/l. The receptor level was similar in both sexes. As compared with the controls, the cellular concentration of beta-adrenergic receptors was higher in eight regularly competing long-distance runners (73 +/- 4 fmol/mg protein; p less than 0.025), whereas the corresponding receptor level in six sprinters (62 +/- 9 fmol/mg protein) did not differ from that of controls. The concentration of lymphocyte beta-adrenergic receptors in seven patients undergoing supine immobilization for weeks because of neck fracture or dislocation showed a wide range from 44 to 155 fmol/mg protein.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3035704 TI - Eicosapentaenoic- and arachidonic acid metabolism in isolated liver cells. AB - The oxidation and esterification of 14C labelled eicosapentaenoic acid (20:5,n-3) and arachidonic acid (20:4,n-6) in isolated liver cells has been studied. The bioconversion of C18 and C22 polyunsaturated fatty acids to 20:5(n-3) and 20:4(n 6) is also discussed. Adrenic acid (22:4,n-6) and docosahexaenoic acid (22:6,n-3) are retroconverted to 20:4(n-6) and 20:5(n-3) respectively by peroxisomal beta oxidation. 20:4(n-6) and 20:5(n-3) are both mainly oxidized in the mitochondria. The peroxisomal contribution to the oxidation of 20:5,n-3 is however larger than with 18:2(n-6) and 18:1(n-9) which are predominantly oxidized by mitochondrial beta-oxidation. Isolated liver cells oxidize more 20:5(n-3) and esterify less than observed with 20:4(n-6) as substrate. In liver cells from essential fatty acid deficient animals 20:5(n-3) and 20:4(n-6) are both efficiently directed to the phospholipids. In hepatocytes from animals fed a diet rich in 18:2(n-6) and 18:3(n-3) arachidonic acid is still to a large extent esterified in the phospholipids while 20:5(n-3) only to a small extent is esterified in this lipid fraction. 18:2(n-6) is much more efficiently esterified in the phospholipids than is 18:3(n-3) which is also rapidly removed from the phospholipids. The delta 4 desaturase activity is increased in essential fatty acid deficiency similar to delta 6 desaturase. The competition between 18:3(n-3) and 18:2(n-6) for delta 6 desaturation in combination with a much higher dietary intake of 18:2(n-6) than of 18:3(n-3) may limit the conversion of dietary 18:3(n-3) to 20:5(n-3).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3035706 TI - Production and characterization of human-human and human-mouse hybridomas secreting Rh(D)-specific monoclonal antibodies. AB - Human monoclonal antibodies specific for the Rh(D) antigen were produced by cell lines generated by the fusion of pooled Epstein-Barr virus (EBV)-transformed B cell lines secreting Rh(D) antibodies with the murine myeloma cell line NS.1 or with the human lymphoblastoid cell line HOA.1. The selection of hybrids was achieved in RPMI 1640 medium containing HAT and ouabain. Higher fusion efficiency was obtained with the NS.1 cell line; however, the hybrids with HOA.1 exhibited a greater clonal stability. The products of four clones (three human-human and one human-mouse) that consistently secreted antibodies for over 11 months were tested for specificity with a panel of red cells of various Rh phenotypes. The supernatants of all four clones showed anti-Rh(D) specificity but failed to react with the red cell Du phenotypes categorized as DV(Dw+) and DVI. Two of the three human-human clones secreted IgM(lambda) and the third IgG(kappa). The human-mouse clone produced IgG(kappa) antibody. PMID- 3035705 TI - Multiple molecular forms of rat liver pantothenate kinase. AB - Pantothate kinase (E.C.2.7.1.33) was partially purified from rat liver and analyzed by preparative isoelectric focusing and cation-exchange high performance liquid chromatography. Two main peaks of catalytic activity were found in the focusing gel at isoelectric point of 5.7 (peak A) and 5.1 (peak B). Two major molecular forms of the enzyme was also demonstrated by high performance liquid chromatography. Both molecular forms of the enzyme were inhibited by 0.02 mmol/l CoASH and acetyl-CoA. CoA at 0.02 mmol/l inhibited the isoenzyme peak B only slightly (6%) compared to the isoenzyme peak A (80%). Acetyl-CoA at 0.02 mmol/l inhibited peak A and B with about equal strength (96% and 83% respectively). The relative amounts of these two molecular forms of pantothenate kinase in different organs might contribute to the regulation of CoA synthesis in mammalian cells. PMID- 3035707 TI - A study of cyclic nucleotides as second messengers after interleukin 2 stimulation of human T lymphocytes. AB - Interleukin 2 (IL-2) was shown to induce a small but significant increase in the level of cGMP after 20 min stimulation and a subsequent fall after 1 h in activated T lymphocytes. No change in the level of cAMP was observed. Addition of the cyclic nucleotide analogues dbcAMP or dbcGMP did not stimulate DNA synthesis. On the contrary, IL-2-induced [3H]thymidine incorporation was inhibited by these drugs. Further, the phosphodiesterase inhibitor theophylline inhibited proliferation of activated T lymphocytes. Our results indicate that neither cAMP nor cGMP act as 'second messengers' for IL-2 but support the theory that cAMP is a negative regulator of cell proliferation. PMID- 3035708 TI - Absent latissimus dorsi muscle and anhidrotic axilla in Poland's syndrome. Case report. AB - The case of an 18-year-old female with absence of the right breast, sternocostal part of the major pectoral muscle and total absence of the minor pectoral muscle and the latissimus dorsi muscle, is presented. She also had decreased sweating capacity and absence of terminal hairs in the central part of her right axilla. Histological examination of skin specimens from this area of the axilla demonstrated hypoplasia of the apocrine sweat glands, but normal eccrine sweat glands, indicating an apocrine dependent axillary sweating function. The breast was constructed using a direct subcutaneous prosthesis implant, giving a final result that was good in regard to shape and softness. This provides grounds for reconsidering the use of the latissimus dorsi muscle flap in breast constructions in Poland's syndrome. PMID- 3035710 TI - Extended intrathoracic resection for lung cancer. Follow-up of 49 cases. AB - Extended resection was performed for primary lung cancer (stage III) on 49 patients in 1973-1982. Their mean age was 61 (38-76) years. In addition to pneumonectomy (29) or lobectomy (20), surgery included resection of the thoracic wall (8 cases) left atrium (12), pericardium (15), parietal pleura (13) and oesophagus (1 case). Among the 47 "surgical survivors" (96%), the mean survival time was 19.9 +/- SD 20.3 months, and only four patients (9%) were alive after 5 years or more. The cumulative 5-year survival was 14% (4/29 patients). Poorly differentiated tumour forms (squamous cell cancer) carried the worst prognosis, whereas the type of resection and presence or absence of lymph-node metastases did not per se influence survival. The prognosis in extended resection is poorer than in standard lung resection, but superior to that in simple exploration. The surgeon therefore should always be prepared to extend a planned resection when a patient on the operating table is found to have extension of lung cancer to other intrathoracic organs, since only in invasion of the chest wall is the surgical strategy as a rule clear from the outset. PMID- 3035709 TI - Interrelated effects of nucleoside mono- and triphosphates and phosphoenolpyruvate on rat hearts subjected to cardioplegic arrest at normothermia. AB - Supplementation with phosphoenolpyruvate (PEP) and ATP was previously found to enhance the protective effect of potassium cardioplegia on rat hearts subjected to extensive ischemic trauma (30 min at 37 degrees C) in the paracorporeal rat heart model. In the present experiments, the ischemia time was reduced to 20 min (37 degrees C). Ventricular work after ischemia was best in control rats with potassium cardioplegia only. Supplementing the cardioplegic solution with PEP and ATP (group I) resulted in significantly reduced postischemic ventricular work and increased efflux of the creatine kinase isoenzyme MB (CK-MB). The same result was obtained when adenosine monophosphate (AMP) was added to the supplementation (group II). When guanosine monophosphate (GMP) was added instead of AMP (group III), the negative effects of PEP and ATP in the cardioplegic solution were partly abolished. Plain potassium cardioplegia, without additives, nevertheless gave the best results. There were no significant intergroup differences in the myocardial content of adenine nucleotides. The results contrasted with those in the previous study of more protracted ischemic trauma (30 min at 37 degrees C) and possible explanations are discussed. PMID- 3035711 TI - The ultrastructure of membranes in sympathetic ganglia. AB - This review presents some recent observations made on membranes in the mammalian sympathetic ganglia after application of freeze-fracturing and histochemistry. After freeze-fracture clear differences have been found between the neuronal and the satellite cell plasmalemma. The satellite cell plasma membrane exhibits specialized intramembrane particles not found in the neuronal membrane. Freeze fracture cytochemistry reveals a further difference between the neurons and satellite cells i.e., a higher density of beta-hydroxysterols is present in the satellite cell membrane than in the neuronal membrane. Histochemical methods to localize 5'-nucleotidase, Ca2+-ATPase and Na+/K+-ATPase in the membranes have been utilized. The cytochemical reaction products of 5'-nucleotidase and Ca2+ ATPase were found on the external aspect of the plasma membranes. The Na+/K+ ATPase reaction product was located on the cytoplasmic aspect of the membranes, with most activity seen in the satellite cell membrane. With respect to the intercellular junctions, the presynaptic membranes have been analyzed with the freeze-fracture technique under stimulated and unstimulated conditions and ultrastructural differences have been observed which might be correlated to neurotransmission. Furthermore, gap junctions and tight junctional elements occur between the satellite cells surrounding the neuronal perikarya and their processes. PMID- 3035712 TI - Receptor-mediated binding, endocytosis and cellular processing of macromolecules conjugated with colloidal gold. AB - Receptor-mediated expression of cellular functions assures biological specificity, regulation, and control of nutritive and metabolic requirements. The study of receptor-ligand interactions is a central theme in many published reports employing colloidal gold labeling. With the ligand directly conjugated with colloidal gold, the investigator is afforded the opportunity to observe all phases of cellular binding, endocytosis, lysosomal delivery and catabolism. Reviewed are published studies employing direct ligand conjugation with colloidal gold, the relative merits and disadvantages of this type of procedure and data from recent studies investigating endothelial receptor binding of proteins in the coagulation and fibrinolysis cascades. PMID- 3035713 TI - [Atypical ultrasound picture of pleomorphic adenoma]. AB - Presentation of an atypical sonomorphology in a case of pleomorphic adenoma of the parotid gland. The tumour was resected and worked up for histological examination. The results emphasise the difficulty of differentiating the subtypes of parotid neoplasms by ultrasonography alone. PMID- 3035715 TI - Expression of functional cell-cell channels from cloned rat liver gap junction complementary DNA. AB - An oocyte expression system was used to test the relation between a complementary DNA (cDNA) clone encoding the liver gap junction protein and cell-cell channels. Total liver polyadenylated messenger RNA injected into oocytes induced cell-cell channels between paired oocytes. This induction was blocked by simultaneous injection of antisense RNA transcribed from the gap junction cDNA. Messenger RNA selected by hybridization to the cDNA clone and translated in oocyte pairs yielded a higher junctional conductance than unselected liver messenger RNA. Cell cell channels between oocytes were also formed when the cloned cDNA was expressed under the control of a heat-shock promoter. A concentration-dependent induction of channels was observed in response to injection with in vitro transcribed gap junction messenger RNA. Thus, the liver gap junction cDNA encodes a protein that is essential for the formation of functional cell-cell channels. PMID- 3035716 TI - Derivation of clones close to met by preparative field inversion gel electrophoresis. AB - The molecular analysis of genes identified by mutations is a major problem in mammalian genetics. As a step toward this goal, preparative field inversion gel electrophoresis (FIGE) was used to selectively isolate clones from the environment of genetically linked markers, and to select a subset of these clones containing sequences next to specific restriction sites rare in mammalian DNA. This approach has been used to generate a library highly enriched in sequences closely linked to the cystic fibrosis marker met. One clone derived from the end of a Not I restriction fragment containing the met sequence was analyzed in detail and localized within a long range map to a position 300 kilobase pairs 5' of the metD sequence. PMID- 3035714 TI - Soil to plant transfer of 239 + 240Pu, 238Pu, 241Am, 137Cs and 90Sr from global fallout in flour and bran from wheat, rye, barley and oats, as obtained by field measurements. AB - Crops of wheat, rye, barley and oats were grown on fields where the contamination of the soil with radionuclides resulted exclusively from global fallout debris. After machine harvesting and milling, the concentrations of 239 + 240Pu, 241Am, 137Cs and 90Sr were determined separately in bran and flour, as well as in the soils. The concentrations of 239 + 240Pu in wheat, rye and barley flour were between 59 and 180 microBq kg-1, and in oat flour 1100 microBq kg-1. The range of concentrations of the other radionuclides for the four flours were: 241Am, 5-70 microBq kg-1, 137Cs, 260-380 mBq kg-1; 90Sr, 310-1300 mBq kg-1. The corresponding concentrations of the radionuclides were always considerably higher in the bran than in the flour, with the exception of oats. The range of this factor depends on the cereal: for 239 + 240Pu, 19-25; 241Am, 10-38; 137Cs, 4-6; and for 90Sr, 4 7. Similar differences between flour and bran were observed for stable elements, for example K, Ca and Fe. The soil-to-plant transfer factors of 239 + 240Pu for wheat, rye and barley flour were between 0.00026 and 0.00078 (oats 0.017) and for bran between 0.0048 and 0.02 (oats 0.014). For 241Am the corresponding values were somewhat lower. For the wheat, rye and barley flour the transfer factors of 137Cs were between 0.013 and 0.018 (oats 0.069) and, for the bran, between 0.08 and 0.10 (oats 0.16). The corresponding values for 90Sr were significantly higher: for flour between 0.08 and 0.14 (oats 1.2) and for bran between 0.62 and 0.93 (oats 1.5). PMID- 3035717 TI - Protein-DNA interactions in vivo upstream of a cell cycle-regulated human H4 histone gene. AB - Cell cycle-dependent histone genes are transcribed at a basal level throughout the cell cycle, with a three- to fivefold increase during early S phase. Protein DNA interactions in the 5' promoter region of a cell cycle-regulated human H4 histone gene have been analyzed at single-nucleotide resolution in vivo. This region contains two sites, with four potential protein-binding domains, at which the DNA is protected from reaction with dimethyl sulfate in cells and from digestion with deoxyribonuclease I in nuclei. These protein-DNA interactions persist during all phases of the cell cycle and dissociate with 0.16 to 0.2M sodium chloride. PMID- 3035719 TI - Influence of cytarabine on mitochondrial function and mitochondrial biogenesis. AB - Although replication of nuclear DNA is inhibited by cytarabine (ara-C), protein synthesis in the nucleocytoplasm appears to continue unabated for the duration of at least the time of the normal cell cycle. ara-C treatment of human leukemic cells resulted in increased mitochondrial membrane potential and adenosine-5' triphosphate (ATP) production and increased activity of enzymes, coded on nuclear DNA (citrate synthetase), as well as of enzymes with subunits coded on mitochondrial DNA (cytochrome c oxidase). These mitochondrial changes occurred during a period of cell-cycle arrest, while cell size and cellular protein content continued to increase. These phenomena appeared to precede the ultimate cell death. PMID- 3035720 TI - Deoxycytidine kinase and deoxycytidine deaminase values correspond closely to clinical response to cytosine arabinoside remission induction therapy in patients with acute myelogenous leukemia. AB - In this study, it has been shown that in 21 patients with AML the dCyd kinase and dCyd deaminase activities correspond closely to the clinical response to ara-C remission induction therapy. Patients with primary disease were treated with a conventional-dose ara-C regimen whereas nonresponders and relapsed patients followed an ID ara-C regimen (1 g/m2 X 12). Of these 21 patients (11 with primary disease and ten relapsed), seven had ara-C resistant disease (three primary and four relapsed patients). Five of the seven patients had a very low dCyd kinase and normal dCyd deaminase activity, whereas the other two had a normal dCyd kinase and an increased dCyd deaminase activity. PMID- 3035722 TI - Mucinous adenocarcinoma of the esophagus metastatic to a spinal fusion. PMID- 3035718 TI - The role of individual cysteine residues in the structure and function of the v sis gene product. AB - The v-sis oncogene encodes a platelet-derived growth factor (PDGF)-related product whose transforming activity is mediated by its functional interaction with the PDGF receptor. PDGF, as well as processed forms of the v-sis gene product, is a disulfide-linked dimer with eight conserved cysteine residues in the minimum region necessary for biologic activity. Site-directed mutagenesis of the v-sis gene revealed that each conserved cysteine residue was required directly or indirectly for disulfide-linked dimer formation. However, substitution of serine for cysteine codons at any of four positions had no detrimental effect on transforming activity of the encoded v-sis protein. These results establish that interchain disulfide bonds are not essential in order for this protein to act as a functional ligand for the PDGF receptor. The remaining four substitutions of serine for cysteine each inactivated transforming function of the molecule. In each case this was associated with loss of a conformation shown to involve intramolecular disulfide bonds. These studies provide insight into the role of individual cysteine residues in determining the structure of the sis/PDGF molecule critical for biological activity. PMID- 3035721 TI - Mechanism for ara-CTP catabolism in human leukemic cells and effect of deaminase inhibitors on this process. PMID- 3035723 TI - Silicosis in the manufacture of scouring powder. PMID- 3035725 TI - [Considerations on organizing a follow-up for an adolescent girl]. PMID- 3035724 TI - Ethical considerations of informed consent: a case study. AB - Presentation is made of a case study concerning a patient with chronic obstructive pulmonary disease who was also discovered to have small cell carcinoma. The patient had indicated many years prior to his hospitalization that if he ever had cancer he would not want to receive chemotherapy. He also indicated to his physician he did not want to hear any 'bad news'. The dilemmas posed by the ethical relationship between consultant and primary care physician and between consultant and patient in such a case are explored. The role of a hospital ethics committee in advising the parties involved is also reviewed. PMID- 3035726 TI - [An admission unit for adolescents in a hospital environment]. PMID- 3035727 TI - Community outbreak of adenovirus type 7a infections associated with a swimming pool. AB - In July 1982, an outbreak of pharyngitis caused by adenovirus type 7a occurred among children in a small town in western Oklahoma. Predominant symptoms were fever and sore throat (by case definition), headache (83%), abdominal pain (64%), and conjunctivitis (51%). At least 77 persons were identified whose symptoms met the case definition for illness. Onsets of illness peaked during the week of July 5 to 12, and the outbreak resolved within three weeks. A systematic telephone survey of the town revealed that persons who had swum at the community swimming pool were more likely to be ill than those who had not (P less than .001). A second survey of families with season passes to the pool showed that among swimmers, illness was directly related to average number of hours of exposure to the pool each week (P less than .001 by chi-square for trend). In addition, swimmers who reported swallowing pool water were more likely to be ill (29 of 56, 52%) than persons who did not (ten of 41, 24%) (P = .01). Throat-swab specimens from five of seven ill persons (71%) grew adenovirus 7a compared with one of 12 well persons (8%) (P = .01). The pool chlorinator had reportedly malfunctioned during early July. The outbreak resolved with proper operation of the chlorination system. Swimming pools continue to be a potential source of community-wide outbreaks of adenovirus infections. PMID- 3035728 TI - Effect of ketoconazole on the 11-hydroxylation step of adrenal steroidogenesis. AB - We studied the effect of ketoconazole on glucocorticoid metabolism in three patients before and after a continuous four-hour infusion of ACTH. A single 400 mg oral dose of ketoconazole caused a decrease in serum cortisol concentration, while the concentration of 11-deoxycortisol and its ratio to cortisol increased. The decrease in serum cortisol levels was not accompanied by an increase in plasma ACTH concentration. A four-hour continuous infusion of ACTH resulted in an appropriate elevation of serum cortisol, but with a marked increase in serum concentration of 11-deoxycortisol and its ratio to serum cortisol. We conclude that 400 mg of ketoconazole can decrease cortisol synthesis apparently through a partial block of the 11-beta-hydroxylation step; the observed degree of inhibition may not be sufficient to stimulate the hypothalamic-pituitary-adrenal axis or to cause overt symptoms or signs of adrenal insufficiency; and this partial block of the 11-beta-hydroxylation step becomes more evident during a continuous infusion of ACTH, which can stimulate a normal response of cortisol. PMID- 3035730 TI - [Successful resection of the tracheal bifurcation with right upper lobectomy for a malignant tumor in the carina region]. PMID- 3035729 TI - Interpretation of magnetic resonance images making use of in vitro examinations of spinal tissue. AB - T1 and T2 relaxation times were determined in vitro at 21 MHz (0.5 T) for a variety of spinal and paraspinal tissues. Intensity formulas for spin echo and inversion recovery sequences were derived and used to calculate the intensities of these tissues as they would appear in magnetic resonance images. The intensity was calculated as a function of various repetition, echo, and inversion times. It is shown that the combination of acquiring in vitro relaxation time values and calculating intensity as a function of pulse timings is useful to predict the parameter setting for optimal contrast between certain tissues without applying series of magnetic resonance images. PMID- 3035731 TI - [Congenital familial adenomatosis]. PMID- 3035732 TI - An experimental study of vascular damage in estrogen-induced embolization. AB - Temporal sequences of estrogen-induced embolization were studied in both in vivo and in vitro models. Infusion of the estrogen compound into rat mesenteric artery caused local spherocytosis and severe rapid degeneration of endothelial cells, followed by injury to the underlying muscle cells and fibroblasts. These changes were regarded as mainly the results of the embolization effect of conjugated estrogen. A study of cultured vascular endothelial cells suggested narrow margins of effective drug concentrations for cell damage. Also, nuclear disintegration with relative sparing of cytoplasmic constituents seemed to be characteristic of estrogen-induced cell damage in both in vivo and in vitro models. PMID- 3035733 TI - [Combination radiochemotherapy in the limited-disease stage of small-cell bronchial carcinoma. Results following high-dosage radiotherapy]. AB - From 1980 through 1984, 72 patients with small cell bronchial carcinomas were treated by radiotherapy. Twenty-one patients who were in a limited disease stage (LD) received chemotherapy according to the ACO scheme and high-dose radiotherapy with 50 Gy. These patients had a complete remission rate of 67% and a partial remission rate of 33%. The median control period was 37 months. At the time of diagnosis, patients had a median survival time of ten months (six to 36 months). A recurrence in the irradiated region was seen in none of the patients, however, 14% of patients developed a recurrence beyond the irradiation field. The results are compared to literature and discussed. PMID- 3035734 TI - An anatomic approach to tumors of the psoas major muscle. AB - The technique of excision of the entire psoas major muscle via the retroperitoneal route is described. This approach prevents direct seeding of the peritoneal cavity with tumor cells and provides relatively easy access to the entire psoas major muscle and its blood supply. The anatomic characteristics of the retroperitoneal space preclude wide surgical margins in whatever approach is used, and in this regard the retroperitoneal approach is probably no more difficult than the transperitoneal approach. The relative ease with which a primary tumor in the psoas major muscle can be excised makes the retroperitoneal approach desirable from the surgeon's point of view. PMID- 3035735 TI - [A new basic drug prospidin in the treatment of rheumatoid arthritis. III. Comparative analysis of prospidin and cyclophosphamide]. AB - A comparative study of the cytostatic drugs prospidin and cyclophosphamide used in equal doses for rheumatoid arthritis (RA) is reported. Clinical and immunologic effects were determined, and the nature and incidence of side effects and complications were compared. Prospidin showed a more pronounced antirheumatic effect, making for a smaller daily requirement of nonsteroid anti-inflammatory agents or hormones in hormone-dependent cases. Unlike cyclophosphamide, prospidin was not associated with severe side effects and complications precluding further use of the drug. Both drugs demonstrated a regulatory effect on RA-associated immunologic disorders. PMID- 3035736 TI - Corticosteroid treatment in pulmonary sarcoidosis: do serial lavage lymphocyte counts, serum angiotensin converting enzyme measurements, and gallium-67 scans help management? AB - Thirty two patients with persisting pulmonary sarcoidosis fulfilling defined criteria for treatment were managed according to a standard clinical protocol. In this an attempt was made to achieve and maintain maximal radiographic and physiological improvement with individually titrated doses of corticosteroids. Lavage cell counts, serum angiotensin converting enzyme (SACE) determinations, and gallium-67 scans were planned at standard intervals but were not used in management decisions. The study analysed serial measurements in relation to changes in the clinical measurements. Twelve patients' radiographs showed complete clearing, seven cleared partially, and 13 had partial clearing with evidence of fibrosis. There was no predictive value in the initial lavage lymphocyte counts or the SACE or gallium measurements. Notably, in seven patients, substantial radiographic improvement was observed when the initial lavage lymphocyte counts were normal. Higher initial lavage neutrophil counts (p less than 0.02), higher initial radiographic profusion scores (p less than 0.02), and lower vital capacity (p less than 0.01) and carbon monoxide transfer factor (p less than 0.05) were related to incomplete clearing. A repeat study of the patients when their radiograph had cleared maximally showed that the levels of lavage lymphocytes, SACE, and gallium tended to fall, but frequently remained raised even in the presence of a normal radiograph or vital capacity or both. On the other hand, however, most of the patients with a normal lavage lymphocyte count showed persisting abnormality of the radiograph, lung function measurements, SACE, and gallium scan (or of at least one of these indices). The interrelationships between changes in clinical indices (radiograph, vital capacity, and transfer factor) and in lavage lymphocyte counts, SACE, and gallium scans showed that concordance was fairly poor in each comparison; lavage lymphocytes showed a greater major discordance than did the other pairs of measurements. Symptom free patients with normal or stable radiographic appearances have been followed for many months and have shown no clinical deterioration despite abnormal lavage lymphocyte counts, SACE, and gallium scans. Radiographic relapse, within the criteria defined, was seen in only four patients during the study; this was reflected in the gallium counts in three and in SACE and lavage lymphocyte counts measurements in two. It is concluded that serial lavage lymphocyte counts, serum angiotensin converting enzyme measurements, and gallium-67 scans are not consistently more sensitive methods by which to monitor patients with sarcoidosis during treatment than are serial measurements of high quality radiographs and results of standard lung function tests. PMID- 3035737 TI - Effect of monocrotaline ingestion on the distribution of protein and angiotensin converting enzyme activity in the rat lung. AB - The alveolar accumulation of protein and angiotensin converting enzyme activity was compared with the development of right ventricular hypertrophy in male rats after different periods of monocrotaline exposure. Total doses of monocrotaline were varied by dividing the animals into three groups in which ingestion was limited to three, seven, and 15 days. These groups were studied 21 days after the start of monocrotaline exposure and compared with a group treated continuously for 28 days. The total lung weight increased after three or more days of treatment, while after seven days there was significant alveolar accumulation of protein, which was paralleled by an increase in angiotensin converting enzyme activity in alveolar lavage fluid. Identical changes also occurred after 15 and 28 days of exposure to monocrotaline. Lung angiotensin converting enzyme activity was decreased after three days' ingestion of monocrotaline and did not alter further with longer periods of exposure. None of these effects of monocrotaline in the three and seven day treatment groups was associated with right ventricular hypertrophy, which occurred only in animals treated for 15 or more days. The effects of monocrotaline ingestion on the lung were dose related and had no causal relationship to the development of right ventricular hypertrophy. PMID- 3035739 TI - PGI3 production from eicosapentaenoic acid in 3T3 fibroblast cells and cultured bovine pulmonary artery endothelial cells. AB - Production of PGI3 from eicosapentaenoic acid (EPA) in 3T3 fibroblast cells and in cultured bovine pulmonary artery endothelial cells (CPAE) which had an ability to produce PGI2 from arachidonic acid (AA), was investigated by bioassay, gas chromatography-mass spectrometry (GC-MS) and thin layer chromatography (TLC). Inhibition of platelet aggregation was observed in the supernatant obtained from a culture medium of 3T3 fibroblast cells after incubation with EPA. This active substance in the supernatant could be identified as PGI3, since the inhibitory effect of the supernatant was blocked by tranylcypromine, a potent inhibitor of prostacyclin synthetase. The inhibitory effect on platelet aggregation by the supernatant from culture medium with EPA was low compared with that with AA. More delta 17-6-keto PGF1 alpha from EPA was produced than 6-keto PGF1 alpha from AA at the same concentration of substrates, when the formation of prostacyclins from EPA and AA was measured as their stable metabolites, delta 17-6-keto PGF1 alpha and 6-keto PGF1 alpha by GC-MS respectively. It is suggested that the anti aggregatory ability of PGI3 is lower than that of PGI2. On the contrary, the amount of PGI3 produced from EPA was significantly less than that of PGI2 produced from AA in CPAE. The production of 6-keto PGF1 alpha from exogenous AA was decreased by increasing the ratio of EPA to AA in the culture medium and the production of delta 17-6-keto PGF1 alpha from EPA also decreased by an addition of AA to the culture medium. This result suggests that AA and EPA compete each other at the site of action of cyclooxygenase.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3035738 TI - Cyclic nucleotide phosphodiesterase inhibitors prevent aggregation of human platelets by raising cyclic AMP and reducing cytoplasmic free calcium mobilization. AB - Cyclic nucleotide phosphodiesterase inhibitors (HL-725, RO 15-2041, cilostamide, quercetin and MY-5445) potently inhibit human platelet aggregation induced by ADP. In parallel, PDE inhibitors inhibit the increase in cytoplasmic free Ca2+ evoked by ADP, as measured with the fluorescent probe quin 2. The inhibition of ADP-induced aggregation and rise in [Ca2+]i is potentiated by PGE1 which stimulates adenylate cyclase and is inhibited by adrenaline which inhibits adenylate cyclase. PDE inhibitors increase human platelet cAMP levels in the presence of low concentrations of PGE1. It is suggested that PDE inhibitors prevent platelet aggregation by raising cAMP levels and by subsequent inhibition of cytoplasmic free Ca2+ mobilization. PMID- 3035740 TI - Dose-related stimulation of platelet cyclic adenosine monophosphate by prostacyclin in thrombotic stroke. PMID- 3035742 TI - High- and low-affinity receptors regulate platelet responses to phorbol diesters and teleocidin. AB - We examined binding of 3H-phorbol dibutyrate (3H-PDBu) to gel filtered human platelets (GFP) and discovered that GFP possess two classes of receptors for phorbol diesters (PDE). High-affinity (HA) receptors, approximately 5000/GFP, bound 3H-PDBu with an apparent dissociation constant (KD) of approximately 12 nM. Low-affinity receptors were approximately 5 times more numerous (2.4 X 10(4)/GFP) and had a 10-fold lower affinity for 3H-PDBu (apparent KD = 115 nM). The potencies of phorbol myristate acetate (PMA) and PDBu paralleled their binding affinities to the PDE receptors. Teleocidin (Tel), although structurally distinct from PDE, competed with 3H-PDBu for its HA-receptors (KI Tel = 1.9 nM). Binding of PDE to HA- or LA- receptors was rapid, reversible, saturable and stereospecific. The HA- and LA-receptors modulated different platelet responses. HA-receptors regulated the secretion of beta-thromboglobulin from alpha-granules and the release of N-acetyl-beta-D-hexosaminidases from lysosomes. LA-receptors mediated both platelet aggregation and the release of serotonin from dense granules. This is the first demonstration of two physiologically active classes of PDE/Tel receptors in human platelets, and demonstrates that particular platelet responses may be directed by distinct classes of receptors for specific agonists. PMID- 3035741 TI - Analysis of distribution of receptors among platelets by flow cytometry. AB - We have investigated the binding of fluorescein isothiocyanate (FITC)-labeled fibrinogen to platelets using a fluorescence spectrophotometer and a flow cytometer. The amount of fibrinogen bound by stimulation of adenosine diphosphate (ADP) and 5-hydroxytryptamine (5HT) increased in a dose-dependent manner. Stimulation of one agonist was independent of that of another agonist. Fibrinogen could bind to platelets in response to ADP, regardless of whether they were desensitized by 5HT, and vice versa. It was found by flow cytometric analysis that individual platelets responded uniformly to both ADP and 5HT, indicating that individual platelets have receptors for both ADP and 5HT on their surfaces. PMID- 3035744 TI - [2.5 mg versus 5 mg neostigmine for reversal of pancuronium bromide]. PMID- 3035743 TI - Leukotrienes do not modify the effects of PAF on human platelet aggregation. PMID- 3035745 TI - [Atypical epithelial hyperplasias, ductal and lobular carcinoma in situ. Treatment, follow-up and prognosis]. PMID- 3035746 TI - [Serological studies on swine. Significance and interpretation of laboratory results for clinical practice]. AB - Establishing a diagnosis in swine herds on the basis of previous histories, clinical symptoms and/or pathological lesions is becoming increasingly difficult for veterinarians. The services of a laboratory for performing serological examinations are therefore increasingly enlisted. Some aspects are described in the present paper, which are essential to an optimum serodiagnosis of Haemophilus pleuropneumoniae infections, Aujesky's disease, influenza and parvovirus infections. PMID- 3035748 TI - [Parvovirus enteritis in dogs based on autopsy statistics 1978-1985]. AB - The occurrence of canine parvovirus enteritis was monitored in post mortem record from 1978-1985. In a total of 7,615 canine necropsies 654 cases of parvovirus enteritis were diagnosed. The number of cases rose considerably in 1980, accounting for about 10% of the total of canine necropsies since then. Seasonal distribution of cases shows a maximum in November, December, and January and a minimum in June, July and September. In German shepherds and Yorkshireterriers parvovirus enteritis is diagnosed with significantly higher frequency than in other breeds. 80% of all cases of parvovirus enteritis necropsied were diagnosed up to the age of 6 months. PMID- 3035747 TI - The proteins of equid herpesvirus 1 (EHV 1) recognised by equine antisera and their ability to promote antibody-dependent cell-mediated cytotoxicity. AB - Equine sera were used to immunoprecipitate radiolabelled virus-infected cell proteins; subsequent resolution with polyacrylamide gel electrophoresis identified the EHV-1 polypeptides VP 2, 10a, 11, 13, 14, 15, 16, 20, 21 and 23a. The humoral support of ADCC by these sera was examined in vitro. Cytotoxicity could be demonstrated against both subtypes irrespective of the immunising isolate. The implications of these results are discussed. PMID- 3035749 TI - Antiproliferative assay of human leucocyte interferon. AB - An in vitro microtitration system of the antiproliferative effect of human leucocyte interferon (IFN-alpha(Le)) was investigated. The preliminary experiments suggested that the antiproliferative effect of IFN-alpha(Le) was increased by prolonging the incubation period, by reducing the target cell concentration, and might be prescribed by the total IFN amounts in the challenge medium. By employing a system consisted of 1.5 X 10(4) Daudi cells in 0.2 ml of medium containing IFN dilutions and an incubation period of 3 days at 37 degrees C, the antiproliferative effects of twenty-one lots of partially purified IFN alpha(Le) (PIF-alpha(Le)) preparations were titrated. The interassay variations in the antiproliferative titers of three PIF-alpha(Le) established in the present system were found to be in the same range as those in the antiviral titers estimated by a standard macroplaque reduction assay. The each titration curve was parallel, and the antiproliferative titers, assessed by the reciprocals of the IFN dilutions which suppress the cell growth in 50%, were significantly (p less than 0.01) correlated to the antiviral international units of them. PMID- 3035750 TI - Comparative studies on clinical features and CSF proteins of motor neuron disease patients. AB - Clinical features such as types of diseases, sex ratio, age of onset, sites of initial involvement, the appearance of bulbar signs, and duration of illness were studied in 52 patients with motor neuron disease (MND) with a special reference to immunoglobulin in cerebrospinal fluid (CSF) and serum. Although MND has been thought to be a degenerative disease of unknown cause, our data suggested there are some immunological abnormalities in this disease. The duration of illness and the abnormalities of CSF immunoglobulins appeared to be correlated with the type of disease and the site of initial involvement. However, whether or not these abnormalities in CSF immunoglobulins are directly related to the pathogenesis of MND remains unclear. PMID- 3035752 TI - A method for preparing mouse skin for assessing in vitro dermal penetration of xenobiotics. AB - A method is described for preparing mouse skin for assessment of in vitro penetration of dermally applied compounds. Separation of the epidermis from the dermis was attempted using a dermatome, heat, trypsin and collagenase. When mouse skin was incubated in a collagenase solution and separated using water, the hypodermis and part of the dermis were separated from the epidermis while leaving the hair follicles and hair shafts intact. Morphologically, the skins prepared for in vitro use appeared to offer similar barriers to topically applied compounds as those found in vivo. PMID- 3035751 TI - Cytotoxic protein from Pseudomonas aeruginosa: formation of hydrophilic pores in Ehrlich ascites tumor cells and effect on cell viability. AB - Increased plasma membrane permeability induced by a purified cytotoxic protein from Pseudomonas aeruginosa was studied using mouse Ehrlich ascites tumor cells in incubation medium containing an osmotic stabilizer. In the presence of serum albumin, 40 nM of the cytotoxin was required for cationic imbalance of 2.7 X 10(7) cells per ml at pH 7.4. The rate of passive flux of water-soluble markers with a molecular radius range between 0.3 and 4 nm was used to calculate the size of functional transmembrane pores. Within a short time of intoxication pores of 1 nm in radius were formed. Their stability is inferred from the constant rate of leakage of the most restricted marker during intoxication (60-90 min). The cytotoxin-induced plasma membrane damage led to loss of essential low molecular weight substances and was associated with a decrease of tumor propagation rate in mice. Regression analysis of these functional parameters indicate the reversibility of plasma membrane disorganization up to complete breakdown of the Na+/K+ gradient. The cell can even tolerate partial loss of larger cytosolic compounds under conditions of limited intoxication. PMID- 3035754 TI - [Use of vaccine interferonogens for treating dermato-neurological syndromes of herpetic etiology]. PMID- 3035755 TI - [Micromethod of human interferon titration]. PMID- 3035753 TI - Toxic effects of hypoxia on neonatal cardiac function in the rat: alpha adrenergic mechanisms. AB - Stimulation of peripheral alpha-adrenergic receptors by circulating catecholamines derived from the adrenal medulla is essential for surviving neonatal hypoxia. In 1-day-old rats, where sympathetic innervation of cardiovascular sites has not yet developed, blockade of these receptors results in failure of cardiac performance from a progressive decline in sinus rate and atrioventricular conduction deficits. These effects are absent in 8-day-old rats, where sympathetic efferent innervation has become established. PMID- 3035756 TI - [Purification of recombinant human alpha-N-interferon on an immunosorbent made of serum polyclonal antibodies]. PMID- 3035757 TI - [Purification of human leukocyte interferon by adsorption chromatography]. PMID- 3035758 TI - The effect of conjugated oestrogens (Premarin) on blood loss during periodontal surgery. PMID- 3035761 TI - Differentiation changes in cord blood T lymphocytes induced by synthetic C terminal peptides of thymosin alpha 1. AB - Thymosin fraction 5 and its component thymosin alpha 1 has been reported to play a regulatory role in the later stages of T lymphocyte differentiation. Previous studies from our group have also demonstrated that thymosin fraction 5 can induce changes in purine degradative enzymes and in surface antigenic markers of cord blood lymphocytes, which have been shown to be immature compared with normal adult lymphocytes. Birr et al., have shown that the C-terminal region of thymosin alpha 1 is essential for the biological activity. In this study, the effects of these synthetic peptide fragments on cord blood T lymphocytes were studied. After incubation with different peptide sequences, cord blood lymphocytes were analysed for surface antigenic markers (OKT4/OKT8) and for purine degradative enzymes (PNP, 5 NT). In the range of 1.3 X 10 to 1.3 X 10(-5)M, one of the eleven peptides examined (sequence 13-19) exhibited activity approximately equal to that of the parent peptide thymosin alpha 1: increase in activity of 5'NT (p less than 0.01) and reduction of cells positive for OKT4 (p less than 0.01). Another sequence (20-28) was able to induce an increase in 5'NT only and a third one (20 25) a reduction of cells positive for OKT4. The results obtained in this study confirm that the C-terminal region contains molecular signals for T cell differentiation. PMID- 3035759 TI - Impaired aldosterone secretion from dispersed adrenal capsular cells of chronically alpha-MSH-treated rats. AB - To investigate the chronic effects of alpha-melanocyte-stimulating hormone (alpha MSH) on aldosterone secretion, synthetic alpha-MSH (8 micrograms/day) was infused in Sprague-Dawley rats by miniosmotic pumps for 5 days. Saline was infused in equivalent volume for 5 days using the same type of pumps in the control group of rats. Aldosterone secretion from the capsular cells of the two groups was examined in the basal state and in response to various stimuli of aldosterone secretion. Aldosterone secretion in vitro from the alpha-MSH-treated rats was significantly impaired in response to all stimuli tested including cyclic AMP, suggesting an intracellular defect in aldosterone synthesis in that group. These results are similar to those observed after chronic adrenocorticotropin administration. PMID- 3035760 TI - Contribution of exogenous progesterone to human adrenal cortisol synthesis in vitro: a comparison of early gestation fetal and adult tissues. AB - The ability of human adult adrenal to utilize progesterone (P4) for cortisol (F) synthesis in vitro has been compared with that of fetal adrenal tissue. Explant cultures were studied for 6 days using Ham's F10 medium supplemented with 10% fetal bovine serum (FBS), with or without added P4 as substrate. Short-term (4h) incubations of fresh tissue minces were carried out in Ham's F10, with or without P4, in the absence of FBS. In contrast with the fetal gland, F production by cultured adult tissue was unaffected by addition of P4. During short-term incubations, P4 increased F production 150-fold in the fetal tissue as compared to 2- to 4-fold in the adult. ACTH had no acute effect on the P4 to F conversion in either tissue. These results demonstrate that the fetal adrenal exhibits a greater ability to utilize P4 for F production than the adult adrenal. PMID- 3035763 TI - Reduced activity of erythrocyte, Na, K-ATPase in hypothyroid children. PMID- 3035762 TI - [Genetic transformation of somatic cells. XI. The autonomic replication of the cloned DNA of the bovine papilloma virus in NIH/3T3 mouse fibroblasts and the changes in the growth characteristics of the transformed cells]. AB - Mouse fibroblasts NIH 3T3 were transfected with the plasmid pBPV (142-6) containing full genome of bovine papilloma virus 1, and focuses of morphological transformation were selected 2-3 weeks later. DNA molecules, containing BPV-1 sequences, were isolated from extrachromosomal fraction of transformed clones suggesting stable autonomous replication of BPV in 3T3 NIH cells. In some rescued plasmids deletions spanning E6, 7 genes of BPV were found. It is suggested that these genes are not essential for morphological transformation and autonomous replication in 3T3 NIH cells. BPV-transformed clones are able to grow in the medium containing low concentration (0.5%) of serum. PMID- 3035764 TI - [Polymyositis with polyradiculoneuritis]. PMID- 3035765 TI - The ultrastructural spectrum of malignant fibrous histiocytoma. PMID- 3035767 TI - Adenocarcinoma and large cell undifferentiated carcinoma of the lung. PMID- 3035768 TI - Diagnostic problems in breast pathology: the benefit of ultrastructural and immunocytochemical analysis. AB - This report deals with the diagnostic importance of intracytoplasmic lumina. The separation between tubular carcinoma and sclerosing adenosis, and the ultrastructural features of variants of lobular carcinoma are discussed. The role of electron dense granules as well as other markers for neuroendocrine cells are evaluated, and finally, the difficult separation of both small-cell and spindle cell tumors is considered. PMID- 3035766 TI - Cytoskeletal intermediate filaments: practical applications of intermediate filament analysis. PMID- 3035770 TI - Electron microscopy of histologically difficult to diagnose gynecologic neoplasms. AB - The ultrastructural characteristics useful in identifying a selected series of histologically difficult to diagnose malignant neoplasms of the human gynecological sphere are presented. The results are discussed in relation to routine diagnosis and when appropriate, to their pathogenic origin. PMID- 3035769 TI - Ultrastructure and histogenesis of the renal tumors of childhood: an overview. AB - This review discusses the ultrastructural and immunohistochemical features of the common childhood renal tumors, with an emphasis on their diagnostic usefulness. Speculations regarding their histogenesis also are presented, with the hope that these may serve to diminish some of the confusion surrounding the classification of these morphologically diverse lesions. PMID- 3035771 TI - [The significance of the bladder neck for micturition disorders in young men. Recent functional and pharmaceutical aspects]. PMID- 3035772 TI - Chronic renal failure in Khartoum, Sudan. AB - A study of the clinical presentation and conceivable causes of chronic renal failure (CRF) in 61 Sudanese patients in Khartoum is presented. The clinical features involved almost all the systems, however, gastrointestinal and cardiovascular signs and symptoms predominated. The causes of chronic renal failure in Sudan and Sweden are also studied for comparison. The causes of CRF in Sudan are chronic glomerulonephritis, obstructive nephropathy (stone disease), hypertension and diabetes mellitus in that order. The main causes of CRF in Sweden are chronic glomerulonephritis, diabetes mellitus and chronic pyelonephritis. Of the 61 Sudanese patients 16 have kidney transplants, only one in Sudan, three patients are on regular hemodialysis, nine patients are on intermittent peritoneal dialysis, 16 are on conservative treatment and 17 died during the course of treatment. PMID- 3035773 TI - Transitional cell carcinoma of the urinary tract associated with vulvar Paget's disease: a report of two cases. AB - Two cases of the urothelial cancers associated with extramammary Paget's disease are reported. In one patient, vulvar Paget's disease was discovered 6 years after complete resection of a ureteral tumor. In another patient, vulvar and vaginal Paget's disease developed during a period of repeated transurethral surgery leading up to the final total cystectomy for recurrent bladder carcinoma. Since genital Paget's disease is frequently accompanied by internal malignancies, a skin biopsy is mandatory when an eczematous lesion has been persistent in the genital region of patients with genitourinary cancers. PMID- 3035775 TI - [Bilateral and familial occurrence of germ cell tumors of the testis]. AB - A reclassification study based on the W.H.O. nomenclature was made of 255 testicular germ cell tumours examined in the Department of Pathology of the District Hospital Schwerin between 1958 and 1985. Thereby some cases of bilateral and familial occurrence were found. 4 cases (1.6%) with bilateral tumours were observed. Only in one of the four cases both tumours were discovered at the same time. The longest interval before contralateral occurrence was 19 years. Familial connections were found in 6 cases (2.4%), in one twin-pair and twice in brothers. PMID- 3035774 TI - Fasting gastrinemia and elevated supersaturation with hydroxyapatite of fasting urine--observations in renal calcium stone patients and controls. AB - We evaluated serum gastrin, acid-base status, variables of mineral metabolism in fasting blood, as well as pH, relative supersaturation of stone forming constituents, and crystalluria in the associated fasting urine, of control subjects (n = 12), and in age- and weight-matched male normocalciuric (n = 12) and hypercalciuric (n = 12) patients with idiopathic recurrent calcium urolithiasis (RCU). In RCU, mineral metabolism and acid-base data are unchanged, whereas mean serum gastrin is only insignificantly higher as compared to controls. Subclassification of all participants into categories with either high normal or low-normal gastrin reveals that in RCU with low-normal gastrin there is a higher-than-normal urinary pH and significantly elevated supersaturation of urine with hydroxyapatite. Crystalluria and stone analysis support the assumption that the physico-chemical environment accompanied by low gastrin levels predisposes to urinary precipitation of calcium phosphate with subsequent formation of a stone nidus. pH in fasting urine and integrated fasting serum gastrin correlate significantly, suggesting that low fasting serum gastrin in RCU patients may be considered a risk factor for calcium phosphate stone formation. PMID- 3035776 TI - Status of equine viral arteritis in Kentucky for 1986. PMID- 3035777 TI - Equine influenza in South Africa. PMID- 3035778 TI - Clinical and immunological responses of cats to feline herpesvirus type 1 infection. AB - Cats which were challenged with feline herpesvirus type 1 developed clinical signs typical of feline viral rhinotracheitis whether or not they had been vaccinated against the disease. However, the clinical disease was less severe and of shorter duration in the vaccinated cats. After challenge, feline herpesvirus type 1 was recovered from the nostrils, oropharynx and peripheral blood leucocytes. Leucocytosis, primarily a neutrophilia, occurred initially in all the cats and was followed after clinical recovery by a mild lymphocytosis. Intradermal skin testing with feline herpesvirus type 1 and cell control antigens produced a positive delayed type skin reaction. Histology of the affected skin 72 hours after injection showed cellular infiltration, predominantly with eosinophils and neutrophils. The severity of the reaction was greater and more prolonged in the skin of the ear than in the skin of the abdomen. PMID- 3035780 TI - Effect of human alpha-hybrid interferon on the course of feline viral rhinotracheitis. PMID- 3035779 TI - Preliminary development of a live attenuated canine parvovirus vaccine from an isolate of British origin. AB - Canine parvovirus isolated from a case of haemorrhagic enteritis in a breeding kennel in England was passaged and cloned in cultured feline and canine cells. No significant evidence of pathogenicity was found during six serial passages of the modified virus back through young dogs. The attenuated virus was excreted by inoculated animals and spread rapidly to uninoculated animals held in contact. When high titre attenuated virus was given to the six-week-old offspring of a seropositive dam a prompt seroconversion was observed. When the attenuated virus was used as an experimental vaccine in 108 pups in an infected breeding colony a highly significant improvement was obtained in the accumulated morbidity and mortality compared with a parallel group vaccinated with modified live feline panleucopenia virus. PMID- 3035781 TI - Inhibition of bovine lymphocyte response to phytohemagglutinin by a specific 5 lipoxygenase inhibitor. AB - AA861, a specific 5-lipoxygenase inhibitor, inhibited bovine lymphocyte response to phytohemagglutinin (PHA). Mitogen-stimulated cultures of mononuclear cells produced leukotriene B4 (LTB4) in 24 hours. The production of LTB4 was completely inhibited by concentrations of AA861 that inhibited mitogen-induced 3H-thymidine incorporation. The inhibition of lymphocyte proliferation was totally reversed by LTB4 but not by leukotriene C4 or leukotriene D4. The inhibition of interleukin-2 (IL-2) production by AA861 was also completely reversed by addition of exogenous LTB4 to lymphocyte cultures. Thus, endogenous LTB4 production appeared to be necessary for PHA-induced IL-2 production and lymphocyte proliferation. PMID- 3035783 TI - Physical state of the latent herpes simplex virus genome in a mouse model system: evidence suggesting an episomal state. AB - Herpes simplex virus (HSV) can establish latent infections in tissues of the nervous system. We have examined the HSV-1 genome in both acutely and latently infected mice by CsCl buoyant density gradient centrifugation. Viral sequences, in gradient fractions, were detected by a spot blot technique using nick translated HSV-1 cloned DNA as probe, and mouse chromosomal DNA was located by measuring optical density at 260 nm. Most HSV-1-specific DNA from both acutely and latently infected mouse brains was found to band at the buoyant density of virion DNA. However, some HSV-1-specific hybridization banded at the density of the mouse chromosomal DNA. Further rounds of CsCl density gradient centrifugation of this chromosomal DNA from acutely infected brains released most of the HSV-1 specific hybridizing material, suggesting that the association was due to trapping. In the case of the recycled chromosomal DNA from a latent infection, all the HSV-1-specific hybridization remained chromosomal DNA associated. However, the amount of hybridization was not significantly greater in quantity than the cross-hybridization between HSV-1 and chromosomal DNA from uninfected mice. AW-Ramos, an EBV-transformed cell line containing one integrated copy of the viral genome, was centrifuged under similar conditions and showed little if any shearing; the EBV DNA banded at the density of the host cell chromosomal DNA. We conclude that the majority of the latent HSV-1 DNA exists in an extrachromosomal state in the mouse model. PMID- 3035784 TI - The genetic relatedness of United States prototype bluetongue viruses by RNA/RNA hybridization. AB - The genetic relatedness of the prototype bluetongue viruses isolated in the United States was examined by RNA/RNA hybridization. Genomic dsRNAs of bluetongue viruses were separated by either SDS-PAGE or NuSieve agarose gel electrophoresis and were blotted by both standard Northern technique and the recently developed rapid alkali-blotting method of Li, Kowalik, and Parker (submitted for publication) to positively charged nylon membranes. The blotted RNA was then probed with 3'-end-labeled dsRNA of each serotype. Initially, single segments were individually hybridized to identify cognate genes. Total genomic dsRNAs were then probed to determine genetic relatedness. The genes coding for the nonstructural proteins NS1 and NS2 were the most conserved. Most of the RNA segments coding for the core proteins were also well conserved, with segment S1 showing some diversity among the five serotypes. The outer capsid protein-coding segments demonstrated a wide degree of sequence divergence with segment L2 having little or no cross-hybridization among the five serotypes. PMID- 3035782 TI - Inhibition by suramin of the NADPH oxidase from horse polymorphonuclear leukocytes. AB - Suramin strongly inhibits the NADPH dependent oxidative activity of the plasma membrane fraction from activated horse polymorphonuclear leukocytes. The kinetics of inhibition reveals the existence of one effective binding site located on the inner surface of the plasma membrane with an apparent Ki value for suramin of about 1 mM. Although administration of suramin to intact cells does not affect the immediate respiratory burst, prolonged preincubation in vitro with the drug results in a decrease of H2O2 production without lowering the phagocytic activity of the leukocytes. Possible implications in vivo are briefly discussed. PMID- 3035785 TI - In cell lines constitutively synthesizing a temperature-sensitive ICP4 protein of herpes simplex virus type 1, amount and function of ICP4 are both regulated by temperature. AB - We have established two cell lines that constitutively synthesize a temperature sensitive form of ICP4, the herpes simplex virus immediate-early protein that activates early and late transcription. ICP4 in both cell lines was confirmed to be functionally temperature sensitive when tested by complementation of an ICP4 deletion mutant virus for expression of viral early and late genes. When grown at the permissive temperature the two cell lines contained approximately 5 and 25%, respectively, of the ICP4 present in control HSV-infected cells. If the cells were grown at the nonpermissive temperature, ICP4 levels were reduced by approximately fourfold; a twofold reduction was observed in control cells synthesizing the wild-type protein. The lower levels of ICP4 at the nonpermissive temperature were the result of two effects: a decrease in mRNA which was similar in cells producing the mutant or wild-type form of ICP4 and a more rapid turnover of the protein which was greater for the mutant than for the wild-type form. Our observations of lower levels of ICP4 in producer cells differ from published reports of overproduction of immediate-early proteins at the nonpermissive temperature in human or hamster cells infected with ICP4 temperature-sensitive mutant viruses. This discrepancy may be related to cell species differences since we observed only a modest twofold overproduction of immediate-early proteins at the nonpermissive temperature in infections of mouse cell lines with an ICP4 temperature-sensitive mutant virus. PMID- 3035786 TI - Nucleotide sequence analysis of equine infectious anemia virus proviral DNA. AB - The nucleotide sequence of the integrated form of the genome of the equine infectious anemia virus was determined. By comparison with LTR sequences of other retroviruses, signals for the control of viral gene transcription and translation could be identified in the EIAV LTR. Open reading frames for gag and pol genes were identified and their sequences matched very closely to those determined previously by others. However, in the present study, the pol gene reading frame was open throughout its entire length. The open reading frame for the env gene product was constructed from the sequences of two independent EIAV clones. Thus, a noninfectious genomic-length clone was shown to contain a frameshift mutation approximately in the middle of the presumed env gene coding sequence, whereas the sequence of another clone was open in this region. The deduced amino acid sequences of the EIAV gag and pol products showed closer evolutionary relationships to those of known lentiviruses than to other retroviruses. There was also partial sequence homology between predicted env gene products of EIAV, visna virus, and HTLV-III/LAV. Sequences analogous to the sor region of other lentiviruses could not be identified in our EIAV clone. A short open reading frame at the 3' end of the genome that overlapped env but not the 3' LTR was present but lacked significant sequence similarity to the 3' open reading frames of other lentiviruses. Thus, the sequence and general structure of EIAV most closely resemble those of known lentiviruses. PMID- 3035788 TI - A comparison of different cloned bluetongue virus genome segments as probes for the detection of virus-specified RNA. AB - The seven largest double-stranded (ds) RNA genome segments of bluetongue virus (BTV) serotype 4 as well as genome segment 8 of BTV10 have been cloned into pBR322. The length of the cloned genes indicates that, with the exception of genome segment 1, the entire gene has been cloned in each case. A method is described for isolating different sized cDNA transcripts on alkaline sucrose gradients with very good recovery. The eight cloned genes were compared as 32P labeled probes for the detection of dsRNA from 21 different BTV serotypes. The S1, S3, S4, S5, and S8 genes were identified as being highly conserved. Of these, S5, which codes for nonstructural protein NS1, gave the best hybridization signal with all the dsRNA isolates. All the probes hybridized significantly weaker with the dsRNA of BTV isolates from Australia and Pakistan than with dsRNA from other serotypes. Genome segment 7, which codes for the group-specific antigen P7, was not highly conserved. Even more variation was shown by genome segment 6 which codes for outer capsid polypeptide P5. S2 which codes for protein P2 is the obvious choice for a serotype-specific probe. Hybridization of this probe with dsRNA from other serotypes reflects the cross-neutralization between BTV4 and these serotypes. The hybridization results can also be used to define the relatedness of BTV4 to other serotypes. None of the probes hybridized with dsRNA from any of the other orbiviruses investigated. PMID- 3035787 TI - Partial nucleotide sequence of the Japanese encephalitis virus genome. AB - Approximately 10 kb of the estimated 10.9-kb genome of the Japanese encephalitis virus (JE; Nakayama strain) has been cloned as cDNA; the uncloned portion includes 430 bases at the 5'-terminus and 450 bases at the 3'-end. A map of the genome has been developed through nucleotide sequencing and in vivo expression with the Escherichia coli expression vector lambda gt11 and immunological identification. Sequence results for 4320 nucleotides suggest the JE genome organization is very similar to those of three other flaviviruses for which sequence information is available. Like the other flaviviruses, the JE proteins are encoded by a single open reading frame that continues uninterrupted throughout the region sequenced. Considerable homology exists between the JE RNA and protein sequences and those of the other characterized flaviviruses. Comparative nucleotide and (amino acid) homology values for the M-E-NS1-ns2 segment of JE are approximately MVE, 70% (80%), WN, 68% (76%), and YF, 50% (45%). Even greater homology is suggested when the protein hydrophobicity profiles are compared. The molecular relationships are consistent with the established serological relationships among JE, MVE, and WN viruses and argue that these flaviviruses may have been derived from a common evolutionary ancestor. PMID- 3035789 TI - Dominance of temperature-sensitive phenotypes. II. Vesicular stomatitis virus mutants from a persistent infection interfere with shut-off of host protein synthesis by wild-type virus. AB - The dominance of a mutant (VSV-PI) isolated from a long-term persistent infection over wild-type vesicular stomatitis virus (wt-VSV) is reported. This dominance has some important differences from and similarities to the dominance of conventional ts mutants studied previously (J. S. Youngner, D. W. Frielle, and P. Whitaker-Dowling, 1986, Virology 155, 225-235). Unlike the ts mutants representing complementation groups I and IV, coinfection with VSV-PI does not reduce the yield of infectious wt-VSV at either the permissive (37 degrees) or nonpermissive (39.5 degrees) temperatures. However, in double infections with wt VSV and VSV-PI at 37 degrees, viral RNA synthesis patterns were converted to those of the RNA synthesis phenotype of VSV-PI: reduced mRNA transcription and enhanced replication of genomic RNA. In addition, VSV-PI which shuts off host protein synthesis very inefficiently was able to interfere in double infections with the ability of wt-VSV to rapidly shut off host protein synthesis. This finding suggests that the mutant virus is not just missing the factor(s) responsible for the inhibition of host protein synthesis but has a dominant activity which works in trans to interfere with the shut-off function of wt-VSV. Ultraviolet irradiation of VSV-PI was used to determine the target size of the interference function. The calculated value for the uv target size is equal to that of the viral genome. This suggests that either viral replication or the expression of the last gene on the viral genome (encoding the L protein) is required for interference by VSV-PI with the shut-off of host cell protein synthesis by wt-VSV. PMID- 3035790 TI - DNA processing in temperature-sensitive morphogenic mutants of HSV-1. AB - The anatomy of DNA synthesized by five HSV-1 mutants previously shown to accumulate predominantly empty capsids at the nonpermissive temperature (NPT) was analyzed with Bg/II restriction digestion. At the NPT, all five generated DNA lacking termini, indicating that in the absence of packaging, viral DNA is not processed to unit length. One mutant, F18, was able to process DNA made at the NPT to unit length molecules during a 6-hr period after shift to the permissive temperature. The appearance of unit length molecules correlated with the appearance of staphylococcal nuclease-resistant F18 DNA. PMID- 3035791 TI - Defective interfering particles of human parainfluenza virus 3. AB - A cyclic pattern of virus production was observed when human parainfluenza virus 3 (HPIV3) was serially passaged nine times in LLC-MK2 cells. Viruses produced from serial passages 8 and 9 interfered with the replication of standard HPIV3. Three subgenomic RNA species (DI-1, DI-2, and DI-3) and virus genomic RNA were detected in the progeny virions produced from cells mixedly infected with standard virus and virus from either serial passages 5 or 8. Northern blot analysis with probes representing all six HPIV3 structural protein genes revealed that DI-1 and DI-2 RNAs contain sequences from the 5' end of the standard virus genome. DI-1 RNA contains L, HN, and F specific sequences, while DI-2 RNA contains only L and HN sequences. DI-3 RNA did not hybridize with any of the probes used. The possibility that DI-3 RNA contains sequences from the 5' end of the standard virus genome is discussed. These results demonstrate that 5' defective interfering particles are generated during serial passage of HPIV3. PMID- 3035792 TI - Fractionation of Theiler's virus-infected BHK21 cell homogenates: isolation of virus-induced membranes. AB - A purified fraction containing unique membranes entrapping virions was isolated from homogenates of cells infected with the DA strain of Theiler's virus, after high-speed centrifugation through a sucrose gradient. This fraction, sedimented at 45-50% sucrose, was only found in cells infected with the DA strain but not in cells infected with the GDVII strain of Theiler's virus or in mock-infected cells. Immunogold staining of the membranes entrapping virions, using antivirus IgG antibodies, revealed that the membranes entrapping virions did not incorporate viral capsid antigens. PMID- 3035793 TI - Synthesis of the X-protein of hepatitis B virus in vitro and detection of anti-X antibodies in human sera. AB - A protein of 154 amino acids, predicted to be encoded by the X-open reading frame of the hepatitis B virus (HBV) genome, was synthesized in an in vitro translation system from SP6 transcripts containing the X-coding sequence. As characterized by SDS-PAGE and immunoprecipitation this X-protein possesses the expected molecular weight of 17 kDa and reacts specifically with rabbit antisera directed against a fusion protein from Escherichia coli that contained 145 of the 154 amino acids from the X-sequence. The X-protein, radiolabeled with [35S]methionine, provided a sensitive and specific antigen to screen for anti-X antibodies in sera from HBV patients. Positive signals were obtained preferentially in subjects suffering from HBV-induced liver cirrhosis or primary hepatocellular carcinoma (PHC), i.e., individuals that had been exposed for an extended time period to HBV gene products. Carefully controlled experiments failed to reveal the presence of X related proteins specific to liver specimens from HBV patients. PMID- 3035794 TI - Evidence for a shift in 5'-termini of early viral RNA during the lytic cycle of JC virus. AB - We have used primer extension and S1 analysis to localize the 5'-termini of JC virus (JCV) early RNAs in infected primary human fetal glial cells at various times postinfection and in stable JCV-transformed hamster fetal glial cells. At early times postinfection (Days 1-5), two early transcripts are initiated at nucleotides 5122 and 5082. A major shift in 5'-ends at later times results in the synthesis of a new series of early mRNAs beginning upstream at nucleotide 35 and downstream at nucleotides 5047, 5037, and 5012. In the transformed hamster cells, however, only one RNA species was detected, starting at nucleotide 5122. The mechanism underlying the shift in the initiation site of JCV early RNAs during a lytic infection remains unclear but appears analogous to that which occurs in the SV40 lytic cycle. Since the shift occurs during DNA replication, when T-antigen is at maximal levels, it is possible that T-antigen binding to JCV DNA and/or alterations in chromatin structure contribute to this event. PMID- 3035796 TI - [Quantitative analysis of the cellular ultrastructure of infiltrative breast cancer]. AB - Cataplasia staging was carried out with the aid of a morphometric study of cell ultrastructure of 45 infiltrative breast cancers. A coefficient describing the free polyribosome structure proved to be of the greatest predictive value, its magnitude and cataplasia degree being connected by a direct relationship. Many theoretical errors may be avoided due to application of quantitative methods of electron microscopy. PMID- 3035797 TI - [Realization of reproductive function and hormonally active ovarian tumors]. AB - Reproductive function was studied in 438 women with practically all histotypes of hormone-producing tumors of the ovaries (141-theca cell, 97-granulosa cell, 14 mixed theca-granulosa cell, 11-masculinizing and 26-Brenner tumors). The said function was reduced in 141 reproductive patients (48.8%). It was shown that hormone producing tumors of the ovary do not cause reproductive function to decline. Conversely, it was infrequent or no births in the young age that could have been a factor of tumorigenesis. PMID- 3035795 TI - [The graduated intramuscular ACTH test and the determination of cortisol in saliva]. PMID- 3035798 TI - [Experience in using radionuclide study methods in the gynecological cancer clinic]. AB - The report deals with a retrospective analysis of applications of different in vivo and in vitro methods of radionuclide studies (1863) in patients with gynecological tumors in 1972-1985. The said methods were employed for making primary diagnosis, assessing tumor extension as well as for evaluating the effects of tumor growth and the efficacy of radiation and surgical treatment. Apart from offering considerable advantages as diagnostic procedures, radionuclide studies may be used in planning treatment modalities and schemes of rehabilitation of cancer patients. The results also suggest a wider application of nuclear procedures in addressing the diagnostic problems of practice of oncology. PMID- 3035799 TI - [Histological types of lung cancer and environmental factors]. AB - The incidence rates of different histologic patterns of lung cancer for 1950-1970 and 1981-1985 were compared at 15-30 year-intervals. The relationship between smoking, occupational hazards and distribution of histologic patterns of lung cancer was studied, too. The study failed to establish any correlation between histologic pattern of lung cancer and said factors. The share of certain histologic patterns has remained unchanged within the last 15-30 years. PMID- 3035800 TI - [Prognostic value of urinary levels of corticosteroids and catecholamines in patients with cancer of the transverse colon and rectum]. AB - Long-term results of treatment of colorectal cancer versus levels of 17 ketosteroids, 17-ketogenic steroids, adrenaline and norepinephrine in diurnal urine were evaluated in 116 patients. Normal or slightly elevated urine corticosteroid and catecholamine levels were shown to correlate with good prognosis, particularly, in cases with normal excretion of all (or almost all) hormones studied and their normal ratios. PMID- 3035801 TI - [Radiation treatment of a large ulcerated breast tumor]. PMID- 3035802 TI - [Specific dynamic action of food in obese patients]. AB - Certain features of the body hormonal system participation in the realizing of the specific dynamic action of food (SDAF) were studied in 26 patients with exogenous constitutional obesity. The radioimmunoassay was used to estimate the levels of a number of hormones (gastrin, insulin, glucagon, triiodothyronine, thyrotrophin, adrenocorticotrophin and hydrocortisone) in the peripheral blood of the patients on an empty stomach (the basal level) and 1,2 and 4 hours after food protein intake. The control group consisted of 20 normal subjects. It has been shown that SDAF is a complex process of changing from the state of hunger to satiation that involves many regulatory components, both nervous and humoral. The study of the features of the SDAF development in health and disease presents valuable information on the mechanisms of metabolic disorders in the body in varying pathologic states including obesity. PMID- 3035803 TI - [Practical aspects of using silufol sheets to analyze fat-soluble vitamins]. AB - The authors studied the mobility of the fat-soluble vitamins, A, D, K1 and beta carotene in organic solvents and their mixtures on silufol plates, the influence of the chromatography conditions and the chamber saturation with the solvent vapors on the Rf values of fat-soluble vitamins, as well as the rate of beta carotene tocopherol (vitamin E) and phylloquinone (vitamin K1) destruction under the effect of the day and ultra-violet light. The data obtained are presented. Phylloquinone and carotene proved to be most destroyed, while tocopherol was more resistant under the same conditions. Silufol plates can be used for the control of the production of vitamins, their analysis in varying biological objects, as well as in biochemistry, medicine and pharmaceutics. PMID- 3035804 TI - [Hormonal indices and cyclic nucleotides in duodenal ulcer]. PMID- 3035805 TI - [The Milwaukee syndrome]. PMID- 3035806 TI - [Neuroectomesodermal dysplasia in a child with the clinical picture of incomplete Klippel-Trenaunay syndrome]. PMID- 3035807 TI - [A rare form of Klippel-Trenaunay-Weber syndrome in a 4-month-old child]. PMID- 3035809 TI - [Hepatitis B virus infection and hepatocellular cancer]. PMID- 3035808 TI - Membrane potential of cells and its regulation during aging. 2. Report: the effect of hormones on the level of the cellular plasma membrane polarization. AB - Age-dependent changes in the polarization of plasma membranes (PM) of various cell types and the mechanisms responsible for its regulation were studied in the experiments on the adult (6-8 and old (28-32 months) Wistar male rats. It was found that the effect of the hormones on the PM polarization level is altered during aging. This being related to shifts in the number and affinity of the hormonal receptors, energetic processes and protein synthesis in the cell. PMID- 3035810 TI - [Current significance of infectious diseases. 1: AIDS--current status of knowledge 1986]. PMID- 3035811 TI - [Decreasing atherogenic risks by an eicosapentaenoic acid-rich diet]. AB - (n-3) diets rich in polyunsaturated fatty acids (PUFA) reduce the atherogenic lipoproteins, especially the VLDL (very low density lipoproteins) rich in triglycerides but also the LDL, more effectively than (n-6) PUFA-rich diets. Moreover also other parameters such as high blood pressure and aggregation of thrombocytes are positively influenced, similarly like after (n-6) PUFA-rich diet. Eicosapentaenoic acid (20:5, n-3) has a triglyceride- and cholesterol reducing effect by inhibition of the VLDL-synthesis (apolipoprotein B, triglycerides) in the liver, inhibition of lipogenic liver enzymes, accelerated elimination of VLDL from the circulation, increased excretion of steroids and bile acids into the stools and amelioration of the fat tolerance. The prolongation of the period of haemorrhage and the decrease of the aggregation of thrombocytes is associated with the enrichment of EPA in the platelet membrane. In these cases the decreased thrombocyte-vascular vessel-interaction shall be caused by a changed metabolism of the eicosanoids (secondary products of unsaturated fatty acids with 20 carbon atoms) and eicosanoid-independent mechanisms. PMID- 3035812 TI - [Hypothesis on the existence of an adenoma-carcinoma sequence in the small intestine]. AB - An analysis of a literature survey of 104 adenomas of the ampulla of Vater, 94 of the duodenum and 20 of the jejunum and ileum, as well as 735 carcinomas of Vater's ampulla, 180 carcinomas of the duodenum and 72 carcinomas of the jejunum and ileum, demonstrated, in spite of small case collectives, that there is probably a similar close relationship between adenomas and carcinomas in the small intestine, as in the large intestine. In adenomas of the small intestine signs of malignancy sometimes can be seen, as well as in some case of carcinoma of the small intestine rests of adenomas have been described. The age and sex distribution of the epithelial neoplasms of the small intestine permits an adenoma-carcinoma-sequence. The relative distribution of the adenomas over the different parts of the small intestine corresponds with that of the carcinomas. The adenomas and carcinomas of the small intestine in patients with adenomatosis coli have the same relationship to the neoplasms of the small intestine in patients without adenomatosis coli, as it is valid in the large intestine. The hypothesis of an adenoma-carcinoma-sequence in the small bowel with a great significance, which explains the results best, is therefore proposed. As the distribution of adenomas and carcinomas of the small bowel in patients with and without familial polyposis is equal, the theory is suggested, that the principle of the adenomatosis intestine disease is a general increase of the overall liability to adenomas in the large and small intestine. PMID- 3035813 TI - Olfactory mucosa of Testudo hermanni (Gmelin) (Reptilia: Testudinidae): occurrence of paracrystalline inclusions in supporting cells. PMID- 3035814 TI - Determination of the nucleotide sequence flanking the deletion (0.762 to 0.789 map units) in the genome of an intraperitoneally avirulent HSV-1 strain HFEM. AB - Herpes simplex virus type 1 (HSV-1) strain HFEM which harbours a deletion of 4.1 kbp in its genome (0.762 to 0.789 map units, HpaI DNA fragment P of HSV-1), is apathogenic for mice and tree shrews by the intraperitoneal application route. The exact position of this deletion was determined by DNA sequence analysis. This analysis was performed using the recombinant plasmid pU18HSHF-XmI-B which harbours the flanking genome regions (0.752 to 0.762 and 0.789 to 0.7895 map units) of the deletion in the genome of HSV-1 HFEM, and the recombinant plasmids pU18HSF-XmI-B, pU18HSF-AS, and pHSF-BB-BsH-D, harbouring particular regions of the genome of the virulent HSV-1 strain F at the coordinates 0.752 to 0.761, 0.786 to 0.790, and 0.762 to 0.771, respectively. The comparison of the DNA sequence of this region with the DNA sequences of the corresponding genome regions of the pathogenic HSV-1 strain F and HSV-1 strain 17 showed that the 5' end of the deletion in the genome of HSV-1 HFEM starts at the nucleotide position 3774 of the BamHI DNA fragment B from HSV-1/17. This position is 71 bp upstream of the UL/RL junction of the HSV-1 genome. The 3' terminus of the deletion ends at the nucleotide position 7226 of the BamHI DNA fragment B from HSV-1/17. The position is within the incomplete ninth repetitive box (ACTCC-CACGCACCCCC) and is located 36 bp upstream of the 3' end of the IE 110 mRNA. PMID- 3035815 TI - Varicella-zoster virus infection of human mononuclear cells. AB - Varicella-zoster virus (VZV) DNA was detected in mononuclear cells (MNC) of 7 humans with acute zoster 1-23 days after the onset of skin lesions. To further study the interaction of VZV with human MNC, cells obtained from seropositive normal donors were infected with VZV and analyzed for the presence of viral DNA and proteins. VZV-DNA was detected in T, B, and OKM 1 (monocyte-macrophage) positive cells, and virus-specific proteins were demonstrated by indirect immunofluorescence and immunoprecipitation. Hybridization studies revealed that VZV-DNA did not replicate in human MNC. PMID- 3035816 TI - The analysis of Yaba monkey tumor virus DNA. AB - Yaba monkey tumor virus DNA was analyzed by digestion with the restriction enzymes Hind III, Eco R I, Bam H I, Sal I, and Bgl I. The restriction fragments from 0.7% and 1.4% agarose gels were used to determine an average molecular weight of 95.0 X 10(6). The location of the terminal cross-links was determined using formamide denaturation. Bam H I fragments D and K (molecular weight 9.6 X 10(6) and 3.3 X 10(6), respectively) were shown to be cross-linked. PMID- 3035818 TI - The complete nucleotide sequence of bluetongue virus serotype 1 RNA3 and a comparison with other geographic serotypes from Australia, South Africa and the United States of America, and with other orbivirus isolates. AB - The sequence of the RNA segment 3 of bluetongue virus (BTV) serotype 1 from Australia is presented along with its deduced amino acid sequence. DNA copies of this genome segment were inserted either into the E. coli plasmid pBR322 by homopolymeric tailing or by direct insertion of double-stranded DNA fragments generated by restriction endonuclease cleavage into the appropriate M13 bacteriophage vectors (Vieira, J. and Messing, J., 1982, Gene 19, 259-268). Direct comparisons were made to the nucleotide sequence data of Purdy, M. et al., 1984 (J. Virol. 51, 754-759) and Ghiasi, H. et al., 1985 (Virus Res. 3, 181-190) for the United States of America (US) isolates of BTV, serotypes 10 and 17, respectively. A method for the rapid cloning, sequencing and alignment of orbivirus RNA 3 segments was utilised to compare other geographical isolates of BTV, as well as those of other orbivirus serotypes, in particular, epizootic haemorrhagic disease of deer virus (EHDV) and Warrego. The comparison of this sequence data reveals that BTV isolates can be separated into distinct geographical types which in turn are distinct from the other orbivirus isolates studied. The sequence conservation at the amino acid level for the gene product of RNA3 (VP3) does not enable distinctions to be made amongst the BTV isolates at a geographical level, but does afford easy distinction into the different orbivirus groups. A possible evolutionary schematic is presented for the orbiviruses studied. PMID- 3035819 TI - [Effect of amizil and the ACTH4-10 fragment on the ability of mice to extrapolate the direction of movement]. AB - Mice possessing the ability to extrapolate the direction of movement (100% of correct choices) were injected i.p. with different doses of m-cholinolytic amizil 2 hours before experiment. Doses of 2-5 mg/kg reduced the percentage of correct choices, the adequate solving strategy being replaced by stereotyped unidirectional reactions or stereotyped alternating responses. Doses of 8-12 mg/kg induced "refusals" to solve extrapolation problem. When amizil treated mice were intraventricularly injected with 1 mg/kg of ACTH4-10, their extrapolation ability was restored. This compensatory action of peptide could be mediated by its influence on cholinergic as well as on other neurotransmitter systems. PMID- 3035817 TI - Genetic characterization of JC virus Tokyo-1 strain, a variant oncogenic in rodents. AB - The genome DNA of JC virus Tokyo-1 strain [JCV(Tokyo-1)], a variant oncogenic in rodents, was molecularly cloned directly from the brain of a Japanese patient with progressive multifocal leukoencephalopathy and from tissue culture, and the restriction enzyme cleavage pattern and regulatory sequences were determined. The restriction pattern of the cloned JCV(Tokyo-1) DNA was different from those of JCVs previously reported in the United States and Germany. Also, the arrangement of the regulatory sequence was unique to this strain. Thus JCV(Tokyo-1) can be classified as a new subtype. The relationship between the restriction pattern and the regulatory sequence of JCV(Tokyo-1), and its characteristic oncogenicity, is discussed. PMID- 3035820 TI - [Antistress effect of dimethyl sulfoxide in the rat]. AB - Administration of scavenger of hydroxyl radicals--dimethylsulfoxide (1 g/kg intraperitoneally, daily for 3 weeks) did not lead to any significant changes in animals behaviour in the open field and in visceral functions (arterial pressure, respiratory rate, heart rate) but prevented shifts of these characteristics caused by chronic 3-week emotional-pain stress. In rats injected with dimethylsulfoxide, an increase was observed of superoxide dismutase activity in the brain and blood serum. Molecular mechanisms are discussed of antistress action of dimethylsulfoxide (scavenge of hydroxyl radicals, activation of superoxide dismutase) and possible role of hydroxyl radicals in realization of damaging action of stress on the organism. PMID- 3035822 TI - [Simultaneous occurrence of Wilm's tumor and multiple lung hamartomas in a 15 year-old girl]. AB - The observation of multiple adenofibroleiomyomatous hamartomas of the lung in a 15-year-old girl 7 years after treatment of a Wilms' tumour caused diagnostic difficulties that are discussed. The diagnosis was finally established by thoracotomy and tumour resection. The simultaneous occurrence of Wilms' tumours and benign or malignant tumours points to genetic factors in the development of both tumours. PMID- 3035821 TI - [Serotoninergic effects in the early ontogenesis of the cat are effected in the neocortex by inhibitory interneurons]. PMID- 3035823 TI - [Value of computerized tomography of the thorax in germinal testicular tumors]. AB - Sporadic informations in the literature that by means of CT of the thorax by far smaller intrapulmonary foci can be recognized in comparison to the conventional x ray examination are just to be confirmed. With the help of instances the indication for thorax CT in germinal testicular tumours is presented. The evidence and exclusion, respectively, of pulmonary metastases are of decisive importance actually for the therapy and prognosis in patients with germinal testicular tumours. In this respect our observations confirm the indications for a CT-examination of the thorax in these groups of patients. PMID- 3035824 TI - [Signet-ring cell carcinoma of the urinary bladder]. AB - The signet-ring cell carcinoma of the urinary bladder as a rare histopathological variant of the adenocarcinoma is demonstrated with the help of clinical, cytological and histological findings of a case (29-year-old male) and it is referred to etiopathogenetic as well as clinical particularities which render possible a delimitation from the carcinoma of the epithelium of the urinary tract. Half of all 28 cases published up to now issued from the remaining parts of the urachus, particularly when localized in the parietal region. The from the first deeply infiltrating growth in the wall of the urinary bladder demands larger surgical interventions, in which cases the partial resection of the urinary bladder represents still the most insignificant operative procedure with clearly better prognosis. For improving the therapeutic situation and the dubious prognosis urachus carriers should be operated on in time, whereby for their recognition modern picture-producing diagnostic method such as sonography should be utilized. PMID- 3035825 TI - [Hard tissue closure of the cleft palate using granular hydroxylapatite-ceramic]. PMID- 3035826 TI - [Syndrome shift following resection of a pathologic intestine and construction of an anus praeter: a comparison of ulcerative colitis, Crohn disease, rectal cancer and familial polyposis]. AB - Postoperative changes of symptoms were studied in 347 patients with Crohn's disease, ulcerative colitis, carcinoma of the rectum, and familial polyposis. A semi-standardised psychosocial questionnaire and a list of complaints, according to v. Zerssen, were used as tools of investigation. The incidence of postoperative disease in patients with Crohn's disease and ulcerative colitis, within the first year from surgery, was higher with significance than that recorded from carcinoma and polyposis patients within the same period of time. Problems relating to identification of syndrome shift are discussed in some detail. The need is stressed for postoperative psychosomatic attention to the patients concerned. PMID- 3035827 TI - [Pregnancy protein-1 in the serum of patients with diseases of the breast]. AB - Pregnancy-specific beta 1-glycoprotein (SP1) was assayed by enzyme immunoassay in serum from 32 patients with breast diseases (11 mastopathies, 6 benign tumors and 15 breast cancers). Healthy blood donors were used as control group. There were significant differences between the SP1-serum concentrations of patient group (mastopathies, benign tumors, breast cancer) and the control group (p less than 0.001). The SP1-values of the groups with mastopathies, benign tumors and breast cancer did not differ significantly. Elevated SP1-concentrations were found in 4 of 11 patients with mastopathy, in 3 of 6 patients with benign tumor and in 10 of 15 patients with breast cancer versus 1 of 24 blood donors. With the help of SP1 determination in serum of 5 breast cancer patients before operation and 6 months post operation we did not found a correlation with further clinical course. PMID- 3035828 TI - Kidney lesions in cattle persistently infected with bovine viral diarrhoea virus. PMID- 3035829 TI - [Localization of Mg2+-ATPase in the membranes of the skeletal muscles and myocardium of the frog Rana temporaria]. AB - Using differential centrifugation in sucrose density gradient, from muscles of the frog fractions were obtained which contain fragments of sarcolemma, as well as membranes of T-system tubules and sarcoplasmic reticulum. In isolated membrane fractions, studies were made on the activity of cation-stimulated ATPases (Na+, K+-, Ca2+, Mg2+- and Mg2+-ATPases). Enzymic and electrophoretic analyses showed that the highest content of Mg2+-ATPases is typical of the fractions which are located on the surface of 35% sucrose. The data obtained indicate that Mg2+ ATPase is the enzyme which is specific for the membranes of T-system tubules in skeletal muscles of not only birds but amphibians as well. From cardiac muscle of the frog, membrane fraction was isolated which is similar (with respect to its predominant content of Mg2+-ATPase) to the membranes of T-system tubules. It is suggested that the presence of Mg2+-ATPase in these membranes is a common property of phasic striated muscle fibers in all mature vertebrate animals. PMID- 3035830 TI - [The ability of argiopine to block the glutamatergic synapses in the frog spinal cord]. PMID- 3035831 TI - [Characteristics of work-loss in diseases of the peripheral nervous system]. AB - Diseases of the peripheral nervous system (PNS) rank fourth among all causes of temporal loss of working ability and first in the structure of neurological morbidity causing temporal disability. Statistical service based on the principles of the International Classification of Diseases, Injuries and Causes of Death does not provide the neurologic service with data about loss of the working time due to PNS diseases. In the comprehensive work on the improvement of neurologic care of patients with PNS diseases conducted in Byelorussia an important place has been recently given to the systematic monitoring of the temporal disability rate which decreased by 28.3% over a period of 3 years (1983 1985). PMID- 3035832 TI - [Decimeter radiothermometry in neurologic manifestations of lumbar osteochondrosis]. AB - Using radiothermometry, a new noninvasive method based on recording the thermoradiation of tissues, the authors studied the in-depth temperature of tissues in 106 patients with lumbar osteochondrosis. A radiothermometer adjusted to a wave length of 30 cm was utilized. The accuracy of measurements was +/- 0.1 degree C. The findings showed an elevation of the in-depth temperature of lumbar tissues by 0.3 degree C as compared with the temperature of the mid-thoracic portion of the vertebral column. The elevation was maximal at the site of the involved intervertebral disc imaging. In the tissue of the leg on the side of pain the in depth temperature was 0.4-0.6 degrees C lower. The authors consider that temperature elevation in the lumbar tissue of the vertebral column is a manifestation of the attendant reactive inflammatory process while a decrease in the temperature of tissue on the side of pain is due to the neurodystrophic process with thermal disturbances. Decimetric radiothemometry may be used for the preoperative localization of intervertebral disk hernias. PMID- 3035833 TI - [Combined use of cutaneous electrostimulation of nerve fibers and electrogymnastics of the spine in painful vertebrogenic syndromes]. AB - A combined use of acupunctural cutaneous electrostimulation of neural fibers and electrogymnastics of the vertebral column was tested in 40 patients with vertebral osteochondrosis accompanied by the muscle tonic or compression radical syndrome. Electrostimulation was performed bilaterally in the paravertebral areas at the level of the blocked motor segment, using series of impulses and achieving rhythmical flexing-unflexing movements of a small range in the damaged segment of the spinal column. Positive effect was obtained in all patients with the muscle tonic syndrome and in half the patients with the compression radical syndrome. PMID- 3035834 TI - [Electroneurophysiologic studies of patients with reflex syndromes of cervical osteochondrosis]. AB - In 21 patients with reflex syndromes of cervical osteochondrosis aged 24-63 years the authors studied short latent somatosensory evoked potentials (EP) in response to stimulation of the median nerve, the rate of impulse conduction along the efferent fibers of the ulnar and median nerves determined with the help of the P wave. The findings obtained by both methods showed a bilateral (on the side of pain and contralaterally) decrease in the rate of impulse conduction along the ulnar and median nerves. The study of the somatosensory systems by registration of EP on the side of pain revealed an increased amplitude and diminished latency of the P9 component which reflects manifestations of irritation of cervical cerebrospinal nodes. PMID- 3035835 TI - [Features of the distribution of HLA antigens in patients with neurologic manifestations of lumbar osteochondrosis]. AB - Examinations of the HLA antigens (A and B loci) distribution in patients with neurologic manifestations of lumbar osteochondrosis (NMLO) has revealed a statistically more rare incidence of A11 antigens and a more frequent one, of B7 and B18 ones. A different distribution of the antigens in cases with the major clinical forms of lumbar osteochondrosis was noted. Patients with discogenic lumbosacral radiculitis differed from normal subjects by the same antigens as all NMLO patients. For patients with lumbar ischialgia increased concentrations of B7 and B18 antigens were characteristic, whereas in radicular ischialgias A11 and B15 antigens were less incident and B40 one, more incident. The findings confirm the polygenic nature of hereditary predisposition to NMLO. PMID- 3035836 TI - [Morphologic changes in tissues of the spinal canal in the radicular syndromes of lumbar osteochondrosis]. AB - Study of biopsy material obtained from 612 surgical patients with vertebrogenic diseases of spinal radicles at the lumbosacral level and collation of the morphologic findings with the clinical picture of the disease revealed a correlation between the clinical manifestations and the inflammatory process in the vascularized tissues of the vertebral column. The severity of pain was found to depend on the degree of inflammatory changes in the epidural cellular tissue and spinal membranes. The frequency and severity of inflammatory reactions make it possible to consider inflammation, along with compression and ischemia, an important pathogenetic mechanism of the development, progression and course of the discogenic radicular syndrome and to recommend the inclusion of antiinflammatory drugs in multiple modality treatment of the disease as a method of pathogenetic therapy. PMID- 3035837 TI - [Status of the pituitary-adrenal and cholinergic system of patients with a pain syndrome in spinal osteochondrosis before and after a course of reflexotherapy]. AB - The pituitary-adrenal and cholinergic systems were studied in 57 patients with chronic pain related to vertebral osteochondrosis before and after an acupuncture course. The degree of the therapeutic effect of electroacupuncture (EAP) was found to correlate with the nature of changes in the level of the spontaneous secretion of aldosterone and intensity of the process of acetylcholine (AC) release. Most patients in whom the treatment was followed by the recovery of the physiologic level of the initially depressed secretion of aldosterone and a clear cut decrease in the initially elevated AC concentrations showed good clinical response. In patients in whom the initially elevated levels of aldosterone and AC failed to decline or in whom there was an even greater increase in AC concentrations, the therapeutic effect of EAP procedures was insignificant and short-lived. PMID- 3035838 TI - [Dynamics of clinico-electromyographic indices in reflexotherapy of radicular syndromes of lumbar osteochondrosis]. AB - Using electromyography the authors studied the functional status of the segmentary apparatus of the spinal cord in 58 patients with compression of S1 radicle secondary to vertebral osteochondrosis. The findings obtained included changes in the interferential EMG and reciprocal reflexes and decreased reflectory activity of alpha-motoneurons predominantly on the side of radicular involvement with the simultaneous intensification of gamma-loop effect. Acupuncture was followed by pain alleviation and normalization of the functional activity of the segmentary apparatus of the spinal cord mainly at the expense of this activity reduction on the healthy side. Clinical recovery was observed following the compensation of neuroreflectory activity of the segmentary centres on a new functional level. The advisability of the use of acupuncture in the treatment of the radicular syndrome of lumbar osteochondrosis is justified. PMID- 3035841 TI - [Massive bone loss in the area of the upper anterior teeth due to subgingival elastic bands]. PMID- 3035839 TI - [Value of manual therapy in the complex treatment of patients with neurologic manifestations of cervical osteochondrosis]. AB - The authors propose a method for combined treatment of patients with varying neurologic syndromes of cervical osteochondrosis which includes manual therapy, traction of the cervical portion of the vertebral column and some physiotherapeutic procedures (hydrocortisone phonophoresis and various baths). The use of drugs was limited. The above treatment was given to 330 patients with reflectory and radicular syndromes of cervical osteochondrosis. The results of the treatment proved significantly better as against those observed in patients receiving conventional treatment. PMID- 3035840 TI - [The necessity of taking into account sanogenetic and pathogenetic reactions in the treatment of patients with the neurodystrophic form of lumboischialgia]. AB - The authors have analyzed the results of clinico-instrumental examination of patients with a neurodystrophic form of lumbar ischialgia associated with lumbar osteochondrosis and specified two groups of symptoms: (1) due to pathogenetic manifestations of the disease and (2) due to sanogenetic reactions of the body. PMID- 3035843 TI - [Selective intra-arterial perfusion of the cancerous breast: quantitative study]. AB - Mammary tissue diffusion of a radioactive marker (Pertechnetate-Tc99m), intravenously or selectively and intraarterially injected, has been studied in 15 breast cancers; a considerable increase of that diffusion with co-injected degradable starch microspheres has been demonstrated. A clinical application of these techniques should be soon considered. PMID- 3035844 TI - Malignant cystosarcoma phyllodes in a man treated with polyestradiolphosphate. Case report. AB - Cystosarcoma phyllodes is a rare breast tumor and only five cases have been previously reported in males. A case of cystosarcoma phyllodes in a man treated with polyestradiolphosphate for prostatic carcinoma and by radiation because of breast tenderness is presented. The tumor showed a malignant stroma histologically and the epithelial component was similar to a ductal carcinoma in situ. Electron microscopy revealed several cell types. The possible relationship to radiation and estrogen therapy is discussed. PMID- 3035842 TI - An unusual variant of Klippel-Trenaunay syndrome. Association of total hemihypertrophy, hemimegalencephaly and bilateral extremity enlargement (case report). AB - The case reported is the first description of an unusual form of Klippel Trenaunay syndrome associating total hemihypertrophy to hemimegalencephaly and bilateral extremity enlargement. The patient complaints are related to her lower limb inequality; she presents dizziness, tinnitus and nystagmus which could be unusual consequences of hemimegalencephaly, disclosed at the EEG. Angiodysplasia is mild in regard to the hypertrophy whose cause is discussed. PMID- 3035845 TI - Natural history of precancerous and early cancerous lesions of the uterine cervix. AB - A prospective study of cervical dysplasia cases and control cases matched for age and parity was undertaken in search of factors related to cervical carcinogenesis. Cytologic examination of 66,736 women revealed negative findings in 28.5%, inflammation in 70.3%, dysplasia in 1.4% and carcinoma in 0.1% of the cases. Data on epidemiologic features, cytomorphologic characteristics and serologic findings of antibodies to herpes simplex virus (HSV) were collected for proven cancer patients, dysplasia cases and control subjects. Cancer patients revealed significantly elevated antibodies to HSV as compared to the controls. The analysis revealed a higher proportion of dysplasia cases with an age at consummation of marriage of less than or equal to 15 years as compared to controls, with a relative risk of 1.5 (P less than .05). Similarly, a higher proportion of women with dysplasia had HSV-II-specific antibodies as compared to control women. The relative risk was found to be 1.3, which was not statistically significant (P greater than .05). The Mantel-Haenszel summary relative risk between antibodies to HSV and the two groups (dysplasia cases and controls), adjusted for the age at consummation of marriage, worked out to be 1.38, which was also statistically not significant (P greater than .05). The overall progression rate of dysplasia to malignancy was found to be 11.7% at the end of 54 months (during a total follow-up period of 84 months). Progression to cancer was highest in severe dysplasia cases and less in mild dysplasia cases. The progression rates were also significantly higher in the group of women who revealed antibodies to HSV II. Similar differences in the progression rates were observed with regard to the age at consummation of marriage. PMID- 3035846 TI - Semiquantitative cytochemical estimation of glucose-6-phosphate dehydrogenase activity in benign diseases and carcinoma of the breast. AB - The level of glucose-6-phosphate dehydrogenase (G6PDH) activity was semiquantitatively evaluated in fresh imprints of infiltrative ductal carcinoma, fibrocystic disease and fibroadenoma of the breast. A significantly higher level of G6PDH activity was found in the carcinomas. The results suggest that the estimation of G6PDH activity could be a valuable method for evaluating the cells in benign and malignant breast lesions. It is possible that the intensification of G6PDH activity in carcinomas is a sign of the shift of the carbohydrate metabolism from an aerobic path or that the activity of the pentose shunt is higher because of the increased need for nucleic acid precursors in tissues with faster growth rates. PMID- 3035847 TI - Primary malignant fibrous histiocytoma of the lung. Fine needle aspiration cytologic features. AB - A case of primary malignant fibrous histiocytoma of the lung occurring in a 71 year-old woman is presented. The preoperative aspiration cytology showed a large cell, undifferentiated, malignant neoplasm suggestive of carcinoma. Subsequent histologic examination revealed a primary malignant fibrous histiocytoma. The diagnosis was confirmed by electron microscopic and immunohistochemical studies. Cytologic features of this rare primary pulmonary sarcoma are discussed. PMID- 3035849 TI - The influence of naloxone on exercise-induced increase in plasma pituitary hormones and the subjectively experienced level of exhaustion in healthy males. AB - Opioid peptides seem to play a role as modulators of the pituitary function in man. In the present study, the effect of naloxone on exercise-induced pituitary hormone release and the subjectively experienced level of exhaustion were investigated in nine healthy males. A submaximal work test was performed on two occasions using a bicycle ergometer: 10 min on 50% of maximal working capacity (MWC), immediately followed by 10 min on 80% of MWC. Ten min before exercise, each subject received, in a single-blind randomized order, either a bolus dose of naloxone (100 micrograms/kg) followed by a slow infusion of naloxone (50 micrograms X kg-1 X h-1) or as a control a corresponding volume of the preservatives in the naloxone preparation as a bolus dose followed by an infusion of diluted preservatives. In the control studies, exercise induced a significant increase in GH, PRL, TSH and ACTH. The increase in ACTH was enhanced following naloxone. Naloxone was without effect on exercise-induced changes in GH, PRL and TSH. An increased level of exhaustion was experienced on 80% of MWC during naloxone. It is concluded that opioid receptors with a moderate sensitivity to naloxone are involved in the regulation of the ACTH response to exercise and also influence the subjectively experienced level of exhaustion. PMID- 3035850 TI - Increased creatine kinase activity in pituitary tumours of rat and man. AB - The demonstration that phosphocreatine is used as an energy source by rat PRL secreting pituitary tumours prompted the study of the enzyme creatine kinase in both rat and human pituitary tumours. Rats treated with diethylstilbestrol developed greatly enlarged pituitaries and hyperprolactinaemia. Total creatine kinase was significantly increased and fractionation on diethylaminoethyl Sephadex showed that the 'brain' form was increased, whereas the 'muscle' and mitochondrial forms showed no change. Exposure to large concentrations of oestradiol caused similar changes in creatine kinase which increased over a period of 25 weeks. The total creatine kinase content of a series of human pituitary tumours was highly variable, but the mean value of 183 +/- 46 (SEM) units per gram protein was significantly higher than the mean for normal pituitary tissues (28.4 +/- 2.9). The brain:muscle isozyme ratio was measured in six human PRL-secreting tumours with a mean of 3.47 +/- 0.73, significantly higher than in 'non-functional' tumours (1.57 +/- 0.29) or normal tissue (1.77 +/ 0.28). Three of four GH-secreting tumours had a ratio below 0.6. The highest ratio found (8.66) was in an ACTH-secreting tumour. Previous reports have shown that oestradiol rapidly induces brain creatine kinase in oestrogen responsive tissues. This is not the case with the rat pituitary gland or oestrogen responsive human tumour cells in culture. Chronic oestrogen treatment, however, does increase creatine kinase in the proliferating gland and many human pituitary tumours have increased enzyme activity. These results suggest that the phosphocreatine/ATP system and in particular the brain isozyme of creatine kinase are of particular importance in lactotropes. PMID- 3035852 TI - Effect of naloxone on the development of the positive feed-back action of oestrogen-progesterone on LH secretion in rats. AB - The purpose of this study was to examine the role of opiate peptides in the development of the positive feedback effect of ovarian hormones (Oe-P) on the LH secretion that matures in female rats at about the age of 20-22 days. Oe-P administration at the age of 14 days induced a significant decrease of LH levels. A single injection of naloxone (5 mg/kg) induced a significant release of LH. This release was completely blocked by Oe-P administration. At the age of 20 days, Oe-P did not induce any significant change of LH levels, whereas naloxone increased the serum LH concentration. On the other hand, injection of Oe-P into naloxone-treated rats induced a significant rise in LH that was significantly higher than that observed with naloxone alone (P less than 0.025). Oe-P administration induced a positive feedback effect on LH at the age of 25 days. At this age, naloxone also increased LH levels and a significant potentiation of the LH release in response to Oe-P was observed in a group treated with naloxone. These results indicate that naloxone advances the development of the positive feedback mechanism of ovarian hormones on LH secretion and potentiates this mechanism after its maturation. On this basis it is proposed that the probable inhibitory effect of opiates on the onset of the positive feedback mechanism is related to the well-known participation of the opiate system in the onset of puberty.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3035848 TI - Cytology of experimental mesotheliomas induced with crocidolite asbestos. AB - The cellular features of 12 pleural and 5 peritoneal effusions, derived from experimental mesotheliomas induced with crocidolite asbestos in white rats, are described. The fluids were obtained 11 to 18 months after the introduction of asbestos into the body cavity. The morphologic characteristics of the cytoplasm, nuclei and nucleoli in the neoplastic mesothelial cells were studied using the Pappenheim, periodic acid-Schiff and Smetana stains. Nearly all effusions examined contained numerous normal and abnormal mitoses. Cell configurations suggestive of amitotic divisions were also observed. The study of the morphologic features of mesothelial cells in effusions in experimental asbestos-induced mesotheliomas may contribute to the understanding of the neoplastic transformation of the mesothelium. PMID- 3035851 TI - Uterotonic activity and myometrial receptor affinity of 1-deamino-1-carba-2 tyrosine(O-methyl)-oxytocin. AB - The contractile effect of the analogue 1-deamino-1-carba-2-tyrosine(O-methyl) oxytocin (carbetocin) on isolated myometrial strips from rats was compared with that of oxytocin. The uterotonic activity of the analogue was 15 +/- 3 IU/mg as compared with 438 +/- 41 for oxytocin, however, the response was more prolonged and could not be abolished by washing of the preparation. Despite the low biological activity of carbetocin, its binding affinity to receptors of isolated myometrial plasma membranes was of the same order of magnitude as that of oxytocin. On the basis of the present results it is concluded that a longer half life of the analogue at the receptor compartment may be a contributing factor to the prolonged uterotonic effect observed in vivo. PMID- 3035853 TI - Effects of atrial natriuretic peptide, dopamine, and ouabain on aldosterone synthesis. AB - This study was undertaken to examine the effects of atrial natriuretic peptide (ANP), dopamine, and ouabain, which each are considered to be a kind of natriuretic factor, on aldosterone synthesis in vivo and in vitro. In the in vivo experiments, during infusion of one of the natriuretic factors, ANP (64 pmol X min-1 X kg-1), dopamine (20 nmol X min-1 X kg-1) or ouabain (684 II pmol x min-1 x kg-1), the stimulatory action of angiotensin (20 pmol X min-1 X kg-1) or adrenocorticotropin (ACTH, 7 pmol X min-1 X kg-1) on aldosterone synthesis was investigated in 36 anaesthetized (pentobarbital, 40 mg/kg, iv) female rabbits. In the basal condition, aldosterone synthesis was suppressed slightly by each of the natriuretic factors. The stimulatory actions of angiotensin II and ACTH on aldosterone synthesis were significantly attenuated by pre-treatment with ANP and dopamine. However, ouabain infusion did not induce any changes in the synthesis of aldosterone, 18-hydroxycorticosterone (18-OHB), and corticosterone (B) in either the basal or the stimulated conditions. For the in vitro experiments, rabbit adrenal glomerulosa cells (10(5) cells/tube) were used. The effects of ANP and dopamine on the aldosterone synthesis revealed results identical to those from the in vivo study. However, in contrast to the in vivo study, ouabain completely inhibited the synthesis of aldosterone, 18-OHB and B. The above results obtained in vivo and in vitro suggest that ANP inhibits corticosterone methyloxidase II activity and dopamine inhibits corticosterone methyloxidase I activity. However, from the present study we were unable to specify the action of ouabain on the synthesis of aldosterone and its related substances. PMID- 3035854 TI - Effects of metabolic inhibitors on hormone release, cyclic AMP levels, and oxygen consumption in rat pituitary cells in culture. AB - The metabolic inhibitors antimycin A (2 mumol/l), dinitrophenol (0.5 mmol/l), and iodoacetate (6 mmol/l) were tested for their effects on hormone release, cAMP levels, and oxygen consumption in clonal strains of rat pituitary cells (GH3 cells). Basal release of growth hormone (GH) and prolactin (PRL) was reduced by all three inhibitors, and thyrotropin-releasing hormone (TRH) (1 mumol/l) and K+ (50 mmol/l) stimulated hormone release were blocked. Trifluoperazine, a calmodulin antagonist, inhibited basal GH and PRL release at concentrations up to 30 mumol/l and stimulated above 50 mumol/l. The stimulatory effect of 80 mumol/l trifluoperazine on basal hormone release was eliminated by antimycin A, dinitrophenol, and iodoacetate, whereas the inhibitory effect of antimycin A, dinitrophenol and iodoacetate on basal hormone was not affected by 30 mumol/l trifluoperazine. None of the inhibitors had any effect on the level of cellular cAMP (i.e. intracellular plus extracellular). Oxygen consumption of GH3 cells was blocked by antimycin A, reduced by 25% by iodoacetate and increased by about 100% by dinitrophenol. In contrast, hormone secretion stimulated by TRH and K+ was not accompanied by any measurable alteration in oxygen consumption. Trifluoperazine (greater than or equal to 80 mumol/l) reduced the basal oxygen consumption and blocked the stimulatory effect of dinitrophenol on oxygen consumption. In conclusion, inhibition of the energy generation of GH and PRL-producing cells severely affects the action of secretagogues, although stimulated hormone secretion may not be accompanied by any measurable increase in oxygen consumption. The cellular energy supporting hormone secretion is mostly generated via oxidative phosphorylation. PMID- 3035855 TI - Hepatocellular carcinoma associated with long-term anabolic steroid therapy in two patients with aplastic anemia. PMID- 3035856 TI - [Levels of fibrinogen degradation products (FDP) and the activity of various granulocytic enzymes]. PMID- 3035857 TI - [Effect of selected phosphate esters on inosine-5-diphosphate pyrophosphohydrolase (ITPH) activity in erythrocytes]. PMID- 3035858 TI - Effects of nootropic drugs on brain cholinergic and dopaminergic transmission. AB - High affinity choline uptake (HACU) in the hippocampus and striatal concentration of dopamine (DA) and homovanillic acid (HVA) as measures of the in vivo acetylcholine and DA turnover, respectively, were estimated in male rats, Long Evans, following 6-day administration of various nootropics in clinically relevant doses: piracetam and its derivatives pramiracetam and oxiracetam (100 mg/kg/day), pyritinol (50 mg/kg/day). Piracetam treatment was without effect on HACU, but induced significant increase of HVA in the striatum leaving striatal DA concentration unchanged. On the contrary, pyritinol, pramiracetam and oxiracetam increased HACU, but did not change striatal DA and HVA levels. PMID- 3035859 TI - Peripheral neuropathy associated with plasma cell dyscrasia: a clinical and electrophysiological follow-up study. AB - Thirteen patients with polyneuropathy associated with plasma cell dyscrasia had serial electrophysiological studies. Five patients with monoclonal IgG had motor and/or sensory symptoms of which 4 correlated with slow motor and sensory nerve conduction. The 4 patients with monoclonal IgM reactive with myelin-associated glycoprotein (MAG), had predominantly motor symptoms, demyelination in the nerve biopsy and slow motor and sensory nerve conduction. Four patients with monoclonal IgM without anti-MAG activity had mainly sensory symptoms, axonal neuropathy on nerve pathology and slow or absent sensory nerve conduction. After treatment with plasmapheresis and chemotherapy 9 patients improved clinically and 4 were unchanged. Criteria for electrophysiologic improvement were presence of sensory or motor responses that were absent before treatment, conduction velocity increased by more than 10 m/s and increase of amplitude by more than 100%. Electrophysiological studies showed improvement in 7, were unchanged in 4, and worse in 2. Sensory velocities in ulnar and sural nerves were significantly improved following treatment (P less than 0.002) and the same trend was noted for the sensory velocity in the median nerve (P less than 0.19). We conclude that nerve conduction studies in combination with clinical examinations are useful in documenting the effects of treatment in these neuropathies. PMID- 3035861 TI - Outbreak of coxsackievirus A-14 meningitis among newborns in a maternity hospital ward. AB - During the late winter of 1983, 16 newborns with vague symptoms of failure to thrive, reluctance to feed and a slight rise in body temperature, were found to have meningitis caused by Coxsackievirus A-14. The cerebrospinal fluid showed pleocytosis with polymorphonuclear cells in excess but was otherwise normal. The clinical course was uneventful in all infants, but two of them demonstrated clinical signs of incipient cerebral oedema during the acute phase of the illness. An electroencephalogram (EEG) during the initial course of the disease and at nine months of age was normal in all. During a follow-up period of 2 1/2 years they all developed normally and no sequelae were noted. The presentation also demonstrates the usefulness of Vero cells for the propagation of the responsible virus. PMID- 3035860 TI - Neuronal antinuclear antibody (anti-Hu) in paraneoplastic encephalomyelitis simulating acute polyneuritis. AB - A patient with paraneoplastic encephalomyelitis (PEM) and small cell lung cancer had a clinical presentation of acute polyneuritis. The patient had an antibody (anti-Hu) restricted to nuclei of neurons identical to that reported in patients with subacute sensory neuronopathy and lung cancer. This finding further supports the hypothesis that PEM and subacute sensory neuronopathy are closely related disorders of autoimmune origin. PEM should be considered in patients with small cell lung cancer and clinical features limited to the peripheral nervous system. PMID- 3035862 TI - Evaluation of renal parenchyma in children by DMSA scintigraphy, X-ray computed tomography and intravenous urography. AB - The demonstration of diminished or scarred renal parenchyma in children is often the decisive factor in determining the future management of children with urinary tract malformations. Renal scintigraphy using technetium 99m-labelled dimercaptosuccinic acid (DMSA), computed tomography (CT) and intravenous urography (IU) were used to evaluate the renal parenchyma prior to ureter re implantation in a series of 13 children. Their ages ranged from 5 months to 3 years 8 months. The indication for operation was ureteric reflux in 8 children and distal ureteric stenosis in 5. CT was performed on a Toshiba TCT-80 scanner. Renal scintigraphy was performed 3 hours after intravenous injection of DMSA. Compared with IU, DMSA imaging gave more information about the renal parenchyma in 6 children, gave equal information in 6 and less in 1. Compared with CT, DMSA imaging gave more information in 2 children, was equally informative in 8 and less so in 3. Accordingly, DMSA imaging was judged to be more sensitive than IU and as sensitive as CT. DMSA imaging can be used both for the initial evaluation and for follow-up assessment of renal parenchymal damage. PMID- 3035863 TI - Maintenance theophylline therapy in children: effect on urinary calcium, phosphate and cyclic AMP excretion. PMID- 3035864 TI - The varied clinical and laboratory manifestations of type II Gaucher's disease. AB - An infant of Arab extraction with the Type II form of Gaucher's disease is described. His clinical presentation was unusual because in addition to the extensive neurological involvement and marked hepatosplenomegaly a severe congestive cardiomyopathy and renal tubular dysfunction were present. In addition, marked hypergammaglobulinemia and raised serum angiotensin converting enzyme levels were found. It is suggested that these varied manifestations may be ascribed to the consequences of glucocerebroside deposition within the macrophages of the reticuloendothelial system. PMID- 3035865 TI - [Synthesis and analgesic activity of 3,4-dichloro-N-methyl-N-[trans-2-(1-delta 3 pyrrolinyl)-cyclohexyl]-benzenacetamide hydrochloride]. PMID- 3035866 TI - [The efficacy of tai-ding-an on experimental herpetic keratitis in rabbits]. PMID- 3035867 TI - Antiviral alkaloid in Chelidonium majus L. PMID- 3035868 TI - [Autoradiography of [3H]ohmefentanyl binding with opiate receptors in the rat brain]. PMID- 3035869 TI - [Mode of inhibitory action of gossypol on Na+, K+-ATPase in vitro]. PMID- 3035870 TI - [Changes in adrenal benzodiazepine receptors in spontaneously hypertensive rats]. PMID- 3035871 TI - [Benzodiazepine derivatives: relationship between anti-convulsant activity and affinity to rabbit frontal cortical receptors]. PMID- 3035872 TI - Changes in serum corticosterone and testosterone during induced maternal behavior in rats. AB - Serum corticosterone (Cpd B) and testosterone (T) concentrations were studied during the avoidance phase of artificially induced parental behavior in male rats. Adult rats were exposed to the presence of standard size foster pups for 60 min daily. On day 1 the avoidance behavior characterized by typical burying reaction was accompanied by an elevation of Cpd B and T. The behavioral responses and hormonal changes diminished during the days of repeated exposure. It was found that neither pup-killing nor spontaneous retrieval were dependent on circulating hormones in the male rat. Subcutaneous injection of ACTH 4-10 inhibited extinction of the avoidance and the humoral responses in both female and male animals. Oxytocin failed to exert any influence on the behavioral and hormonal components of the aversive reaction. PMID- 3035873 TI - Influence of increased environmental temperature on oxidation processes in rat liver mitochondria. AB - Exposure of rats to elevated temperature of 28 degrees C or 35 degrees C for 3 days six hours daily resulted in a decreased rate of oxidation with succinate or glutamate + malate as substrates, by the mitochondria of liver. The higher decrease was observed in environment temperature of 35 degrees C. There was no change in ADP/O ratio. The activities of NADH: cytochrome c reductase and cytochrome oxidase were stimulated but activities of succinate dehydrogenase and succinate cytochrome reductase were decreased. PMID- 3035874 TI - Split-respiratory centre in the dog. PMID- 3035876 TI - Evidence for the existence of an endogenous inhibitor of Na, K-ATPase in plasma from cats and rats. AB - Plasma samples from cats and rats were filtered through molecular filters to obtain a concentrate of the plasma molecules with a mass between 500 and 10,000. This concentrate was placed on a gel filtration column and eluted with a Tris buffer. Fractions corresponding to a molecular mass of 1500-2000 exhibited a marked Na,K-ATPase inhibitory 'activity', which was not influenced by heating to 95 degrees C for 10 min or by enzymatic degradation by pronase or trypsin. PMID- 3035875 TI - ACTH-mediated aldosterone hypersecretion during endotoxin-induced fever with apparent influence upon renal sodium excretion. AB - Febrile, endocrine, and renal responses to i.v. injection of endotoxin (E. coli lipopolysaccharide, 0.25 microgram kg-1) were studied in hyperhydrated goats without, and after dexamethasone pre-treatment, performed with the aim of inhibiting the adenohypophyseal secretion of ACTH. As expected from previous investigations, the administration solely of endotoxin induced biphasic fever, pronounced and long-lasting (less than 4 h) elevation of plasma cortisol (PC), and a prompt inhibition of the water diuresis. Apparently the observation that endotoxin also induced a pronounced biphasic elevation of plasma aldosterone (PA) where the two rising phases coincided with the early, and respectively the second elevation in rectal temperature is original. The endotoxin had no obvious influence upon the renal Na excretion for 3 h post-injection, and did not affect plasma renin activity (PRA). After dexamethasone pre-treatment (0.02 mg kg-1, i.v. 75 min prior to endotoxin) the endotoxin-induced rise in rectal temperature initially was less steep and no biphasic pattern of the fever was observed. The PC and antidiuretic responses became delayed for about 2 h and were then much attenuated. Endotoxin-induced rise in PA was no longer observed, and a conspicuous natriuresis developed within 90 min post-endotoxin. It is concluded that endotoxin at the dose used causes liberation of ACTH to such an extent that adrenocortical hypersecretion not only of glucocorticoids, but also of aldosterone occurs. The observed differences in Na excretion suggest that this aldosterone hypersecretion may be of pathophysiological importance as a protection against inappropriate renal waste of Na during the early phase of endotoxin-induced fever.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3035877 TI - Effects of N-ethylmaleimide and forskolin on noradrenaline release from rat hippocampal slices. Evidence that prejunctional adenosine and alpha-receptors are linked to N-proteins but not to adenylate cyclase. AB - N-ethylmaleimide (NEM) treatment has been shown to inactivate regulatory GTP binding N (G)-proteins in many preparations, including slices of rat hippocampus. NEM-treatment (100 microM for 15 min) has been used to examine the possible involvement of a N-protein in the prejunctional inhibitory effect of an adenosine analogue, R-PIA acting on A1-receptors, and of clonidine acting on alpha 2 adrenoceptors in this tissue. NEM treatment significantly enhanced basal overflow of [3H]NA and the overflow stimulated by low (0.3 Hz) frequency stimulation, but not the overflow stimulated by higher (1-10 Hz) frequency stimulation. The prejunctional inhibitory effect of R-PIA (1 microM) on NA release, stimulated by a 3 Hz stimulation, was abolished by NEM pretreatment, which also eliminated the dose-dependent prejunctional effect of clonidine and reduced the facilitatory effect of yohimbine. Forskolin had a small, but significant stimulatory effect on NA overflow, but did not reduce the prejunctional inhibitory effect of R-PIA. The adenylate cyclase inhibitor SQ 22, 536 did not reduce NA overflow. These results show that NEM blocks a critical step in the prejunctional action of both adenosine- and alpha 2-receptor agonists, which may be a N-protein. The possibility is discussed that the prejunctional A1- and alpha 2-receptors couple to a N-protein that controls a different effector than adenylate cyclase. PMID- 3035879 TI - Postmenopausal osteoporosis: no effect of three years treatment with 1,25 dihydroxycholecalciferol. AB - The therapeutic effect of 1,25-dihydroxycholecalciferol (1,25(OH)2D3) in postmenopausal osteoporosis was tested in a single blind, randomized prospective study. Thirty-nine women, 50-65 years of age, were treated for three years with 0.5 microgram 1,25(OH)2D3 daily. In a control group, 37 women were given 400 IU vitamin D3 daily. There was no significant difference in annual bone loss from the distal or proximal forearm between the groups. New vertebral fractures were evaluated, and in the treatment group, the annual increase in vertebral fractures was 0.18 +/- 0.387 and in the control group 0.13 +/- 0.330. New long bone fractures were 7 and 5, respectively. None of the observed differences were statistically significant. In the 1,25(OH)2D3 group, 28% had to reduce the dose because of slight hypercalcaemia. We conclude that 1,25(OH)2D3 as used in this study is not effective in the treatment of osteoporosis. PMID- 3035878 TI - Comparison between a serotonin and a noradrenaline reuptake blocker in the treatment of depressed outpatients. A cross-over study. AB - Seventy-five outpatients with major depressive disorder (RDC) were randomly referred to treatment with a dominant serotonin (5-HT) reuptake blocker (zimeldine, 100 mg, b.i.d. n = 40) or a dominant noradrenaline (NA) reuptake blocker (maprotiline, 75 mg, b.i.d. n = 35). Seven patients on each drug were non responders after up to 4 weeks of treatment and were after a washout week crossed over to the other drug for up to another 8 weeks of treatment. There was a significant and similar improvement after 4 weeks of treatment with the second drug. After up to 8 weeks of treatment all patients but one in each group were much improved with the second drug. The existence of two biochemical subgroups of depression is discussed. PMID- 3035880 TI - Structure and function of platelet-derived growth factor. PMID- 3035881 TI - Functional domains of apolipoprotein E and apolipoprotein B. PMID- 3035882 TI - Intracarotid infusion of ACNU and BCNU as adjuvant therapy of malignant gliomas. Clinical aspects and critical considerations. AB - Thirty patients with malignant gliomas were treated by operation, radiotherapy and additional intracarotid infusions of ACNU and BCNU. Positive results were obtained in the treatment of oligodendrogliomas and astrocytomas grade III and IV. On the contrary, the results in cases of glioblastoma multiforme were disappointing: neither survival time nor quality of life had been significantly improved. The protective effect of phenobarbitone against systemic toxicity by ACNU was not always confirmed in this study. Based on literature reports and our own experience the indications, technical aspects, unexpected complications and results of this therapeutic approach are discussed. PMID- 3035883 TI - 1986 aspects of steroid hormone receptors: nuclear localization, 8S-hetero oligomeric form, heat-shock protein component and antihormones. PMID- 3035884 TI - The behavioral physiology of vasopressin and oxytocin. PMID- 3035885 TI - The hypothalamic-pituitary-adrenal axis in depression. PMID- 3035886 TI - Corticotropin releasing hormone: relevance to normal physiology and to the pathophysiology and differential diagnosis of hypercortisolism and adrenal insufficiency. AB - CRH is a 41 amino acid peptide first isolated from ovine and subsequently from rat and human hypothalami. We have conducted a series of clinical studies with oCRH and hCRH in volunteers and patients with various disorders of hypothalamic pituitary-adrenal function. In volunteers, it was demonstrated that hCRH administration produced ACTH and cortisol responses which closely mimic naturalistically occurring secretory episodes. This data, as well as the demonstration that pulsatile hCRH can reestablish normal ACTH and cortisol secretion in patients with hypothalamic CRH deficiency, strongly argue that CRH is of physiological relevance to the human pituitary-adrenal axis. However, since the ACTH response to an insulin tolerance test is greater than the maximal ACTH response to CRH, other factors such as vasopressin may be relevant to stress induced ACTH secretion in man. Following the demonstration that CRH seems to be of physiological relevance to human subjects, a CRH stimulation test was developed based on pharmacokinetic and dose response studies with oCRH and hCRH. Based on these data, which revealed that oCRH functions as a long-acting analogue of hCRH, and the demonstration that hormonal responses to CRH are greatest in the evening, patient groups with abnormalities of the hypothalamic-pituitary-adrenal axis were tested with intravenous oCRH with a dose of 1 micrograms/kg given at 2000 hours. This CRH stimulation test has proved helpful in clarifying the pathophysiology of hypercortisolism in a variety of psychiatric disorders characterized by this endocrine abnormality. Thus, blunted ACTH responses in hypercortisolemic patients with depression, anorexia nervosa, and panic anxiety disorder indicate normality of the pituitary corticotroph in these patient subgroups. These data, along with the finding that a continuous infusion of CRH to normal volunteers, reproduces the pattern and magnitude of hypercortisolism in depression and anorexia nervosa, suggest that the hypercortisolism in these disorders represents a defect at or above the hypothalamus resulting in the hypersecretion of CRH. This hypothesis is particularly intriguing in light of the demonstration that CRH administration to experimental animals produces many of the physiological and behavioral responses classically associated with depression and anorexia nervosa, including hypercortisolism, hypothalamic hypogonadism, and decreases in libido and appetite. The CRH stimulation test has also helped to resolve one of the oldest endocrinological dilemmas, namely whether the hypercortisolism of depression and Cushing's disease share a common or dissimilar pathophysiological basis.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3035887 TI - Opioid peptides at term pregnancy in the early puerperium and in postpartum psychosis. PMID- 3035888 TI - Hypothalamic functions, sleep and circadian rhythms in affective disorders. PMID- 3035889 TI - Human monocyte chemotaxis to neuropeptides. PMID- 3035891 TI - Endogenous opioids and hypothalamic-pituitary-adrenal function in obesity and anorexia nervosa. PMID- 3035890 TI - Neuroendocrine measures in anorexia nervosa: comparisons with primary affective disorders. PMID- 3035892 TI - ACTH-related peptides, kindling and seizure disorders. AB - Very small amounts of pituitary hormones and their peptide fragments can profoundly affect learning, memory and other behaviors in both rodents and humans. In addition, several potent pituitary hormone analogs have been developed (e.g. ORG-2766) which retain the behavioral but not the endocrine properties of the parent hormone. The abilities of these peptides to influence nervous system functions suggested that they also may be capable of modifying seizure activity. This possibility is supported by the fact that treatment with adrenocorticotropic hormone (ACTH) represents a major effective therapy for at least one clinical convulsive disorder, infantile spasm. This contention also is reinforced by our findings that ORG-2766 markedly reduces both the behavioral severity and the spread of seizure potentials in an animal model of epilepsy, the kindled rat. By contrast, arginine vasopressin and its non-endocrine desglycyl fragment (DGAVP) facilitates the seizure process in this animal model. Our research also suggests that the behavioral and physiological effects of certain anterior and posterior pituitary hormone fragments depend, in part, on their abilities to modulate permeability mechanisms in brain vasculature. In addition, and especially significant from the standpoint of etiology, is the observation that the kindling process itself appears to alter cerebrovascular permeability. In the kindled rat, "permanent" decreases in permeability (60-75%) are found selectively in the hypothalamic and hippocampal regions, weeks after the last seizure. Several lines of evidence indicate that disruption of normal cerebrovascular function occurs following epileptiform seizures. We propose that an immediate increase in cerebral blood flow and cerebrovascular permeability occurs following a single, acute seizure, and that repeated chronic seizures lead to damage to the cerebrovascular system. Sustained damage would be expected to contribute to the development and maintenance of a chronic seizure focus. Such observations suggest a link between important cerebrovascular disturbances associated with seizures and the existing known and proposed electrophysiological, metabolic and neuropathological substrates of epilepsy. They also point to new strategies for the treatment of seizure disorders by focusing on ways to reduce or prevent cerebrovascular damage. PMID- 3035895 TI - Biosynthesis of neuropeptides in the rat central nervous system. PMID- 3035893 TI - Neuropeptides and seizures: an experimental model on the possible relationship among C.R.F., ACTH and endorphinic system. PMID- 3035894 TI - Brain receptors for hypothalamic hormones. AB - Angiotensin II and CRF are but two of the several regulatory peptides which exert specific actions in the brain that are complementary with their peripheral effects upon end organs such as the anterior pituitary and adrenal glands. In the pituitary, the two peptides act in a coordinate manner on the corticotroph to regulate ACTH release. In the adrenal gland, angiotensin II receptors are abundant in the zona glomerulosa but are also present in the medulla, where the occurrence of CRF receptors and actions on catecholamine release reveals an additional site at which the two peptides exert related actions, in this case in the peripheral neuroendocrine system. Within the brain, the mapping of AII and CRF binding sites by topical autoradiography has provided new information about the distribution and potential functions of receptors for the two peptides. The central receptors for AII are distributed in a characteristic pattern in brain regions concerned with drinking, regulation of adrenergic function and arterial blood pressure, and control of pituitary hormone secretion. Thus, in addition to its recognized modulatory effects in the peripheral adrenergic system, angiotensin II may be involved in the central control of catecholamine release and action. A central action of AII on the release of regulatory peptides such as vasopressin and CRF, both of which are present in neurones of the paraventricular nucleus, is indicated by the high concentration of AII receptors in this region. Also, the high density of AII receptors in the median eminence suggests that AII modulates the hypothalamic secretion of neuropeptides such as CRF by actions at their site of release, as well as on the cell bodies of neurones responsible for peptide synthesis. The highly localized pattern of AII receptors at numerous specific sites in the brain differs from the more general distribution of many other CNS receptors, and reflects the selective actions of AII on discrete neural systems that subserve precisely integrated functions within the central nervous system. The widespread distribution of CRF receptors, with prominent localization in the cortical and limbic regions, is consistent with the more general neuroregulatory actions of CRF in the brain, and with the presence of immunoreactive CRF in several regions of the brain including the cortex, limbic system, and centers involved in the control of autonomic function. The cortical and limbic receptors are clearly relevant to the effects of centrally administered CRF on both behavioral and visceral responses, with prominent autonomic changes including increased catecholamine release and hypertension. PMID- 3035896 TI - Modification of experimental colon carcinogenesis by dietary fibers. AB - The literature concerning the effect of individual dietary fibers on the experimental induction of colorectal cancer was reviewed. It has become increasingly apparent that the effect of dietary fibers on colon carcinogenesis depends on many factors, including the type and amount of fiber; the other dietary components, particularly fat; animal species, strain, and sex; and the type of carcinogen and its dose and route of administration. Despite such variations in design, most experiments with wheat bran and cellulose have shown evidence of a significant protective effect. In contrast, numerous other fiber supplements have been shown to enhance tumor development. These include pectin, corn bran, undegraded carageenan, agar, Metamucil, and alfalfa. Possible mechanisms by which fibers may inhibit colon tumorigenesis include dilution and adsorption of any carcinogens or promoters contained within the intestinal lumen and faster transit time and therefore less opportunity for carcinogen/promoter interaction with the intestinal epithelium. Modulation of colonic microbial metabolic activity by dietary fibers may also be important in the activation and detoxification of carcinogens and promoters. Dietary fibers produce structural and functional changes in the intestinal epithelium and modify rates of cell proliferation changes in the intestinal epithelium and modify rates of cell proliferation and migration. Evidence suggests that if this stimulus to cell proliferation occurs during the stage of initiation, it may lead to enhancement of the carcinogenic process. Dietary fibers bind not only carcinogens, bile acids, and other potentially toxic agents but also essential nutrients that themselves can modify the carcinogenic process. Fermentation of fibers within the large bowel results in production of volatile fatty acids, which in vitro have been shown to be antineoplastic. Fermentation also produces a lower luminal pH, which in turn affects colonic microbial populations and their metabolic activities. The presence of lignans in higher plants and their bacterial synthesis from precursors present in fiber-rich foods provide an additional source of antineoplastic agents, whose relative importance in colon carcinogenesis is unknown. Because dietary fibers differ in their physiochemical properties, it has been difficult to identify a single mechanism by which fibers prevent or inhibit colon carcinogenesis. Clearly, more investigation is needed regarding the mechanism(s) by which certain fibers inhibit while others enhance experimental colon carcinogenesis. PMID- 3035897 TI - Dietary fiber and human cancer: critique of the literature. AB - The relationship between dietary fiber consumption and risk of gastrointestinal cancer in humans is examined using representative studies of several types: international and intranational correlations, case-control analyses, metabolic investigations, cohort studies, and migrant studies. The strongest statistical association between diet and cancer is found in international studies in which numerous environmental variables differ. Studies on smaller groups within a single culture have not given strong support to the findings of international comparisons. Colon cancer rates within regions of the U.S. and other countries vary with sufficient magnitude that diet is unlikely to account for more than a minor proportion of risk. The evidence that a diet containing fiber-rich foods reduces risk of colon cancer must be considered tentative. Foods high in starch and fiber are statistically associated with a high rate of stomach cancer. Examination of the combined rates of colon and gastric cancer shows that the U.S. risk is low relative to countries in which a diet higher in fiber is consumed. It would be premature to suggest that a high fiber diet will confer protection against gastrointestinal cancer. PMID- 3035898 TI - Lipotropic factors and oncogenesis. AB - The lipotropes (choline, methionine, folate, and vitamin B12) have a rich history, with many fluctuations in scientific effort and popularity, covering the past 6 decades. A thin thread of common interest in 1-carbon metabolism and a small band of dedicated individuals have kept this area of biology alive. Today, the lipotropes are enjoying a resurgence of interest and effort with promise for significant contributions to some of our most serious chronic diseases. Between 1920, when Banting and Best initiated a work that led to the discovery of insulin, and 1982-83, when investigators working in 3 laboratories announced that lipotrope deficiency alone could result in liver cancer in rodents, many have used this model to study nutritional problems and, more recently, carcinogenesis. Lipotropes are important to lipid metabolism and to synthesis and maintenance of cellular membranes. When weanling rats were fed a diet low in lipotropes, within a few days the liver accumulated lipid, first in the centrilobular zone and later throughout the entire lobule and lobe. If the diet was continued for a longer period, the liver underwent fibrosis and cirrhosis with some rats ultimately developing hepatocellular carcinoma. Although lipotrope deficiency can result in liver cancer, all hepatocarcinogens tested thus far were enhanced in their activity by diets low in lipotropes. Important changes associated with lipotrope deficiency included membrane damage, decreased serum very low density lipoprotein and drug metabolizing enzymes, decreases in S-adenosylmethionine and in methylation of cytosine, increases in cellular peroxidation products and free radicals, decreased immunocompetence, and a markedly shortened lag time for chemical induction of liver cancer in animals. The overall effect of lipotrope deficiency is an increase in the susceptibility to cancer in animals; the exact mechanisms are unclear. PMID- 3035899 TI - Selenium, vitamin E, fiber, and the incidence of human cancer: an epidemiologic perspective. AB - It has been hypothesized that selenium, vitamin E, and fiber reduce the risk of specific human cancers. Evidence for a role of selenium is based primarily on animal studies, inverse geographic correlations between intake and site-specific cancer incidence, and an inverse association between serum selenium and subsequent risk of cancer. Certain geographic areas with high fiber intakes have lower rates of colon cancer and, in several case-control studies, consumption of fruits and vegetables has been associated with a lower risk of large bowel cancer. Suspicion that vitamin E might reduce the risk of human cancer is largely theoretical; a protective association has been observed in only 1 small study of breast cancer. The evidence that these 3 dietary factors reduce the risk of human cancer remains incomplete. Future epidemiologic investigations should simultaneously assess a wide variety of dietary factors to address potential confounding and interacting effects. Prospective study designs should be used whenever possible to avoid any influence of cancer on dietary intake or its measurement. PMID- 3035900 TI - The relationship between the vitamin D system and cancer. AB - The classic function of 1,25-dihydroxyvitamin D3, the hormonally active form of vitamin D, is the maintenance of normal levels of calcium and phosphorus in the blood. 1,25-Dihydroxyvitamin D3 binds to a specific receptor protein and exerts its biologic action by a mechanism analogous to that proposed for other steroid hormones, that is, the receptor-ligand complex acts on the chromatin to induce transcription of specific genes. Intracellular receptors that bind 1,25 dihydroxyvitamin D3 with high affinity have been found in a large number of tumor cell lines examined as melanoma, osteosarcoma, and human breast and colonic carcinoma cells. The 1,25-dihydroxyvitamin D3 receptor in these cells has characteristics similar to the receptor in bone and intestine, the known target tissues of the hormone. In fact, 1,25-dihydroxyvitamin D3 inhibits the proliferation of melanoma, osteosarcoma, and breast carcinoma cells. More recently, 1,25-dihydroxyvitamin D3 has been shown to suppress the growth and induce monocytic differentiation of murine and human myeloid leukemia cells in vitro. These results point to a previously unsuspected involvement of vitamin D in cell proliferation and differentiation and suggest that analogs of the vitamin D hormone may be of interest as possible therapeutic agents in the treatment of malignancy. PMID- 3035902 TI - HDL subfractions, HDL receptors and HDL turnover. PMID- 3035901 TI - Alcohol and cancer. AB - The cancers consistently associated with ingestion of alcohol, the head and neck cancers, are also associated with tobacco use and arise from epithelia that are in direct contact with both agents. Tobacco smoking-related cancers at sites not directly in contact with alcoholic beverages, that is, lung, bladder, and perhaps pancreas, do not consistently show a relationship to alcohol consumption, although lung and pancreatic tumors are associated in some studies. Liver cancer was thought to be strongly related to alcohol consumption on epidemiological grounds and because of its relationship to cirrhosis. As knowledge of the viral etiology of some cirrhoses has evolved and as methods to detect viruses have developed, the significant association between hepatitis B virus and hepatocellular carcinoma has become clear. Alcohol and hepatitis B virus may interact in the etiology of the disease and have important separate roles as well. There are epidemiologic and experimental data showing that malnutrition (resulting from poor food choice), economic deprivation, or alcoholism contributes to the risk for head, neck, and liver cancers. Colon cancers occur about equally in men and women, are found in well-nourished populations, and are not associated with tobacco smoking. Rectal cancers show a preponderance of cases in men but are frequently found in women as well and are not thought to be associated with smoking or malnutrition. The association between colorectal cancers and alcohol consumption, when it is found, apparently occurs at even relatively low alcohol intakes and is often stronger for consumption of beer than of other beverages. Nutritional and metabolic mechanisms proposed for the influence of alcohol on carcinogenesis are supported by studies in human subjects and laboratory animals. Animal models are needed in which effects of ethanol on carcinogenesis can be consistently demonstrated and which can then be used to examine mechanisms. PMID- 3035903 TI - Cultured fibroblast interactions with LDL and HDL from healthy subjects on various dietary fats. PMID- 3035904 TI - Psychoneuroendocrine strategies. PMID- 3035905 TI - Regulation of translation by poliovirus. PMID- 3035907 TI - [Wilms' tumor in an adult patient: a case report]. AB - A case of Wilms' tumor in an adult patient is presented. The tumor in adults is rather rare and has an unfavorable prognosis. A 39-year-old man bruised his left flank while skiing in February, 1983. He noticed severe left flank pain. The pain subsided after a week of rest. However, he complained of left abdominal mass and dull pain again, and consulted our outpatient clinic on May 21, 1983. Left transperitoneal nephrectomy was performed under a diagnosis of left injured renal tumor. Histological diagnosis was nephroblastoma. A combined chemotherapy of actinomycin D (ACD) and vincristine (VCR) was started after operation. A total of 10 mg of ACD and 14 mg of VCR was administered by the end of 1983. In March 1984, however, a local recurrence and pulmonary metastases of the tumor were detected by CT and chest films. Local RF-hyperthermia combined with irradiation, alpha interferon or chemotherapy using cisplatin and adriamycin was given. The patient died of profound cachexia in December, 1984 after gradual deterioration of general conditions. PMID- 3035908 TI - Imaging in acute renal infection in children. PMID- 3035906 TI - Flavivirus replication strategy. PMID- 3035909 TI - Uptake of 99mTc-methylene diphosphonate by pancreatic insulinoma. PMID- 3035910 TI - Plasma concentrations of alpha-human atrial natriuretic polypeptide and cyclic GMP in patients with heart disease. AB - Plasma concentrations of immunoreactive alpha-human atrial natriuretic polypeptide (i alpha-hANP) and cyclic guanosine monophosphate (cGMP) were measured in 70 patients with heart disease. Plasma concentrations of i alpha-hANP were directly related to the severity of heart disease (F = 29.61, p less than 0.001). Plasma concentrations of i alpha-hANP were well correlated with pulmonary capillary wedge pressure (PCWP; r = 0.64, p less than 0.001), mean pulmonary arterial pressure (PAP; r = 0.62, p less than 0.001), and mean right atrial pressure (RAP; r = 0.75, p less than 0.001). Plasma concentrations of cGMP were also directly related to the severity of heart disease (F = 13.61, p less than 0.001) and highly correlated with plasma concentrations of i alpha-hANP (r = 0.73, p less than 0.001). Plasma concentrations of cGMP were also closely correlated with PCWP (r = 0.69, p less than 0.001), mean PAP (r = 0.61, p less than 0.001), and mean RAP (r = 0.60, p less than 0.001). The i alpha-hANP concentrations of plasma samples obtained from the coronary sinus were approximately fourfold higher than those of samples obtained from the pulmonary artery, whereas cGMP concentrations were comparable in plasma samples obtained from either site. Elevation of cGMP concentrations following intravenous infusion of synthetic alpha-hANP was comparable in plasma samples obtained from the coronary sinus and the pulmonary artery. These findings suggest that elevated plasma concentrations of i alpha-hANP in cardiac patients result from an increase in the secretion of ANPs, which is probably accelerated by elevation of right or left atrial pressure, and that plasma concentrations of cGMP reflect circulating levels of alpha-hANP. PMID- 3035911 TI - Evaluation of the phase contrast microscopy method for the detection of fibrous and other elongated mineral particulates by comparison with a STEM technique. AB - The USPHS/NIOSH Membrane Filter Method is used to monitor for asbestos in occupational and mining atmospheres, and employs the phase-contrast optical microscope (PCM) that under optimum conditions has a resolution of approximately 0.25 micron. While amphibole cleavage fragments are usually visible by PCM, asbestos fibers (such as amosite and chrysotile) have finer widths that may render them invisible by PCM. In this study, personal air-monitoring filters containing chrysotile, amosite and amphibole cleavage fragments from various sources have been analyzed by PCM in accordance with the USPHS/NIOSH Method and scanning transmission electron microscopy (STEM) to assess the effectiveness of the PCM technique. Each STEM specimen was prepared using a direct-transfer technique to ensure that particle size distribution and concentration were not altered. STEM results for chrysotile samples are highly variable, with 9% to 81% of regulatory particles having widths smaller than 0.25 micron--the resolution of the optical microscope. Amosite samples have 27% to 38% of regulatory particles with widths below microscope resolution, indicating that routine particle counts by PCM on these samples would underestimate true fiber content by approximately one-third. All amphibole cleavage fragment samples had regulatory particles that would be observed by PCM. Multiplication factors have been suggested for application to routine counts by PCM to more accurately assess true particle content for mineral particulates on personal air-monitoring filters. PMID- 3035912 TI - cis-platinum, 5-fluorouracil, and hydroxyurea in the treatment of advanced non small-cell lung cancer. A phase II study. AB - Sixteen patients with advanced non-small-cell lung cancer were treated with the combination of cis-platinum, 5-fluorouracil, and hydroxyurea. There were two complete responses (lasting 16 weeks each) and one partial response (lasting 12 weeks). Toxicity was manageable, but responders were unable to tolerate further therapy. Despite the obtaining of complete responses, the overall benefit of this particular combination requires further clinical investigation. PMID- 3035913 TI - A randomized study comparing platinum, doxorubicin, and VP-16 with platinum, 4' epidoxorubicin, and VP-16 in patients with non-small-cell lung cancer. AB - Forty-four previously untreated patients with advanced non-small-cell lung cancer were treated in a randomized trial comparing platinum (60 mg/m2), doxorubicin (40 mg/m2), and VP-16 (150 mg/m2) (PAV) with platinum (60 mg/m2), 4'-epidoxorubicin (50 mg/m2), and VP-16 (150 mg/m2) (PEV). The overall response rate was 10%. Major response rates were quite similar for the 21 patients treated with PAV (5%) and the 23 patients treated with PEV (18%) (p = 0.2). Of the 23 patients with assigned to PEV, two (9%) achieved complete responses for a median duration of 20 weeks and 44+ weeks. There was no significant difference (p = 0.75) in the median survival among patients treated with PAV (24 weeks) and those treated with PEV (20 weeks). Toxicity was generally mild and tolerable. The lack of response found in both arms of treatment caused the study to be terminated early. Some benefit could be appreciated in patients with limited disease and good Karnofsky performance status. PMID- 3035914 TI - Simultaneous evaluation of terminal deoxynucleotidyl transferase and myeloperoxidase in acute leukemias using an immunocytochemical method. AB - The classification of acute leukemia is important for the selection of optimal therapy. Classification often rests on morphologic, cytochemical, and immunologic criteria, and the marker enzyme terminal deoxynucleotidyl transferase (TdT) has been considered to be a reliable indicator of lymphoblastic leukemias. Because TdT-positive cells sometimes are seen in leukemias otherwise identified as myeloblastic, the authors evaluated blasts identified as myeloid by the presence of myeloperoxidase (MPO) for the simultaneous expression of TdT. The blasts in the bone marrow aspirate or peripheral blood of unselected patients with hematologic malignancies were evaluated and 60 cases are shown. The French American-British system and, in some patients, cytochemical and immunologic studies were used to classify the leukemias. The authors demonstrated that blasts simultaneously contained MPO and TdT in 29% of patients with acute myeloblastic leukemia and 3% of patients with acute lymphocytic leukemia (ALL). This finding supports the hypothesis that TdT is an expression of cell primitivity rather than a marker for lymphoblastic cells. PMID- 3035915 TI - Direct in situ hybridization for rapid detection of cytomegalovirus in bronchoalveolar lavage. AB - Rapid detection of cytomegalovirus (CMV) from pulmonary specimens in immunosuppressed persons may provide an origin for pneumonia. In situ DNA hybridization has been effective for detection of CMV in otherwise nondiagnostic histologic material. Studies comparing bronchoalveolar lavage (BAL) with open lung biopsy have shown the former to be superior in detecting most pulmonary pathogens affecting immunocompromised patients. Fifty consecutive BAL specimens were studied to compare direct in situ DNA hybridization, routine tissue culture, and conventional cytologic examination to assess the efficacy of the hybridization technic to rapidly detect CMV. Using tissue culture as the standard, a sensitivity of 90% (28 of 31) and specificity of 63% (12 of 19) were observed with the CMV probe. Discrepant results between the probe and tissue culture were present in ten cases. There were seven probe-positive, culture negative cases, three of which had systemic CMV infection, including two patients with inclusions noted by conventional cytologic examination. Three probe negative, culture-positive cases were found. In the authors' laboratory, the predictive value of a positive CMV probe is 80% (28 of 35). In contrast to the probe, conventional cytologic examination revealed CMV inclusions in only 23% (7 of 31) of the culture-positive cases. An average of 21 days was required for CMV cultures to become positive; probe results were available within 24 hours. The authors conclude direct in situ DNA hybridization is a useful rapid method for the detection of CMV in BAL specimens submitted for cytologic examination. PMID- 3035916 TI - Pneumocystis carinii involvement of the bone marrow in acquired immunodeficiency syndrome. AB - A case of Pneumocystis carinii involvement of the bone marrow in acquired immunodeficiency syndrome is reported. The literature is reviewed for other cases of known extrapulmonary dissemination of Pneumocystis. Potential mechanisms of dissemination of Pneumocystis and its clinical implications are discussed. PMID- 3035917 TI - Recurrent wheezing episodes following parainfluenza virus bronchiolitis. PMID- 3035918 TI - Phenotypic features of patients with congenital adrenal hypoplasia and glycerol kinase deficiency. AB - Two unrelated boys with congenital adrenal hypoplasia and glycerol kinase deficiency were found to have similar features, including characteristic facies, testicular abnormalities, short stature, psychomotor retardation, and muscular dystrophy. The resemblance of these boys to other patients described in the literature suggests that a distinct phenotypic syndrome occurs in children with congenital adrenal hypoplasia and glycerol kinase deficiency. PMID- 3035919 TI - Poliomyelitis in Spain, 1982-1984: virologic and epidemiologic studies. AB - The authors present a detailed study of poliomyelitis in Spain for the years 1982, 1983, and 1984. The 50 cases reported have been epidemiologically classified following World Health Organization guidelines. Virus was isolated from 43 of these cases. Intratypic characterization was done using specifically absorbed antisera classifying the strains as non-Sabin-like and Sabin-like. The neutralizing antibodies in whole and fractionated sera were also determined. The largest number of paralytic poliomyelitis cases, 28, was found in children in the first year of life. Thirty of the cases were unvaccinated children. Poliomyelitis was not detected in adults during the period described. Wild strains isolated were type I and III, primarily from Gypsies. There was a significant incidence of vaccine-associated cases in recipients, as well as in contacts during 1982 and 1983. The cases were located in the Mediterranean and Southern zone. In 1984, total and vaccine-associated cases dropped dramatically. PMID- 3035921 TI - Gastrointestinal endocrine tumors. PMID- 3035920 TI - Prenatal diagnosis of sickle hemoglobinopathies: the experience of the Columbia University Comprehensive Center for Sickle Cell Disease. AB - We report here an evaluation of 55 pregnancies at risk for a sickle hemoglobinopathy prenatally diagnosed by restriction-endonuclease analysis, with the endonucleases MstII and HpaI, of amniocyte DNA. The diagnosis was completed in all cases. Eleven fetuses were predicted to be affected, of which six were terminated. Forty-one of the 55 cases were confirmed. One false-negative was reported in a case predicted to be hemoglobin AS but that was determined to be hemoglobin SS at birth. We estimate that the 55 cases represent only 5% of the pregnancies at risk for a sickle hemoglobinopathy in the New York metropolitan area during the study period. We conclude that the prenatal diagnosis of sickle hemoglobinopathies by molecular methods is reliable. However, the efficiency of utilization and effectiveness of prenatal testing is dependent on the early prospective identification of couples at risk and on the education of communities concerning the significant morbidity of the sickle hemoglobinopathies and the reproductive choices now available to them. PMID- 3035922 TI - VIPoma syndrome. AB - Since the description of the watery diarrhea syndrome by Verner and Morrison 29 years ago, clinical and experimental observations have elucidated the pathophysiology of this disease. Vasoactive intestinal polypeptide (VIP) is produced and released by a tumor of the pancreatic islets or by a tumor of neural crest origin such as a ganglioneuroma. Under normal conditions, current evidence suggests that VIP is a neurotransmitter in the central and peripheral nervous systems and particularly in the peptidergic nervous system. The low VIP plasma concentration observed in healthy subjects is viewed as a neuronal overflow since it has been impossible to ascertain any endocrine role for circulating VIP. Markedly elevated VIP plasma levels in the VIPoma syndrome lead to intestinal secretion with severe secretory diarrhea, resulting in hypovolemia, hypokalemia, and acidosis. These symptoms subside after successful tumor removal. Approximately 50 percent of patients have metastatic spread at the time of diagnosis. For these patients, a new and promising therapeutic modality is available in the form of a subcutaneously administered somatostatin analogue that relieves symptoms through potent inhibition of VIP release from tumor tissue. PMID- 3035923 TI - Surgery for gut hormone-producing tumors. PMID- 3035924 TI - Chemotherapy of metastatic carcinoid and islet cell tumors. A review. PMID- 3035925 TI - Human cytomegalovirus infection of human placental explants in culture: histologic and immunohistochemical studies. AB - The induction of human cytomegalovirus infection in human first-trimester placentas was studied with a placental explant culture model. Replication and/or release of human cytomegalovirus in placental explant cultures did not occur at any time from 1 to 10 days after infection when examined by plaque assay and analyses of extracted deoxyribonucleic acids. In contrast, typical human cytomegalovirus-induced histopathologic lesions bearing human cytomegalovirus antigens were consistently localized in the trophoblastic cells covering placental villi. These data clearly demonstrate that placental cells are permissive of latent and/or abortive human cytomegalovirus infection in vitro. Our results support the hypothesis that during human cytomegalovirus infection of pregnant women, maternal viremia or intrauterine infection results in latent human cytomegalovirus infection of placental cells that may persist during the course of pregnancy. PMID- 3035926 TI - Angiotensin-converting enzyme activity in hypertensive subjects after magnesium sulfate therapy. AB - Angiotensin-converting enzyme is a peptidase involved in the formation of angiotensin II and the inactivation of bradykinin. In a previous study we found elevated angiotensin-converting enzyme activity in women with pregnancy-induced hypertension prior to magnesium sulfate therapy and lower levels during therapy. This prospective study was undertaken in order to determine if angiotensin converting enzyme activity indeed decreased after magnesium sulfate therapy. Sixteen patients with pregnancy-induced hypertension were studied before and during magnesium sulfate therapy. Angiotensin-converting enzyme activity was found to decrease 1 to 8 hours into therapy and then plateau between 9 and 24 hours. Possible mechanisms for this observation are discussed. PMID- 3035927 TI - Estrogen therapy arrests bone loss in elderly women. AB - Although osteoporosis is an age-related disorder, the accelerated bone loss observed in postmenopausal women may be preventable with early diagnosis and adequate estrogen replacement. In a prospective study, we investigated the effect of oral estrogen replacement using conjugated estrogens (Premarin, 0.625 mg) or micronized 17 beta-estradiol (Estrace, 1 mg) versus no estrogen in sequential single-photon bone density measurements over 3-year intervals in 397 postmenopausal women. Estradiol, 1 mg, and conjugated estrogens, 0.625 mg, were equally effective in regarding bone loss. The rate of bone loss was about the same for estrogen users regardless of age (51 to 80 years) and was approximately one third that of nonusers. Among nonusers a uniform accelerated rate of bone loss of 2.5% per year was noted between 56 and 70 years old, whereas between the ages of 51 and 55 years and after age 70 years, the rate of bone loss was significantly less. Ever users over age 65 years showed continued protection from bone loss as long as estrogen therapy was continued. Previous estrogen users who stopped estrogen after age 65 years lost bone more rapidly than women of similar age who had never taken estrogen. Thus to prevent accelerated bone loss in postmenopausal women, we recommend early and continued hormone replacement for life. Estrogen nonusers should be monitored at regular intervals to minimize accelerated bone loss. PMID- 3035928 TI - Use of intravitreal ganciclovir (dihydroxy propoxymethyl guanine) for cytomegalovirus retinitis in a patient with AIDS. PMID- 3035929 TI - Detection of cytomegalovirus infection by means of DNA isolated from paraffin embedded tissues and dot hybridization. AB - A specific and rapid method for detecting the presence of cytomegalovirus (CMV) DNA in formalin- or mercuric chloride-fixed, paraffin-embedded tissue has been developed. With dot hybridization with a biotinylated CMV DNA probe, the presence of CMV infection can be readily detected even if only a minority of the cells in the tissue block are infected. The sensitivity is further increased by about two orders of magnitude if high specific-activity 32P-labeled probe is used. This method of detection is more sensitive than histologic diagnosis, appears to be roughly comparable to that by virus isolation in culture, and should be applicable to the detection of other infectious agents. PMID- 3035930 TI - Experimental evidence for a neural origin of Ewing's sarcoma of bone. AB - The histogenesis of Ewing's sarcoma remains unknown. Recent studies have suggested a relationship to an unusual form of childhood neural tumor, often termed peripheral neuroepithelioma or primitive neuroectodermal tumor. Five Ewing's sarcoma tumor cell lines were studied for evidence of a neural phenotype. Under normal culture conditions, no morphologic evidence of neural differentiation was detected. Treatment with retinoic acid, an agent known to induce marked neural differentiation in neuroblastoma, had no demonstrable effect. Treatment with either cyclic AMP or TPA, in contrast, induced pronounced morphologic evidence of neural differentiation. Cells developed elongate processes with varicosities by phase-contrast microscopy; filaments, microtubules, and uraniffin-positive dense core granules were present by electron microscopy. Three neural markers (NSE, NFTP, and cholinesterase) were absent or barely detectable in untreated cells, but became abundant after treatment. These results provide convincing evidence for a neural histogenesis of Ewing's sarcoma. They also suggest a close relationship between Ewing's sarcoma and peripheral neural tumors, including the chest wall tumor described by Askin, but only a distant relationship to neuroblastoma. PMID- 3035932 TI - The role of acute and latent virus infections in the pathogenesis of inner ear disturbances. AB - The possible role of herpesviral infections of the inner ear in suddenly appearing inner ear disturbances was investigated. Experimental pseudorabies virus (PRV, Herpes sui 1) infection of mice and swine was used as a model system. Infected mice represented the productive cycle of PRV infection (acute phase), whereas the latent phase of infection could be tested in swine. From the acutely infected mice the virus could be reisolated from perilymphatic fluid and various parts of the brain. Massive histopathologic alterations and signs of total cell damage to the organ of Corti and the vestibular organ were found. Accordingly, in all of the cells of the inner ear multiple copies of the PRV genome could be demonstrated. We therefore suggest that the disturbances of the inner ear were induced by the acute virus infection. In two latently infected swine (sixty weeks after infection), PRV could not be recovered either from the perilymphatic fluid or from a variety of different neural and extraneural tissues. However, histopathologic changes similar to those found in the acutely infected mice were observed. The presence of viral DNA could be demonstrated by in situ cytohybridization in both sensory and supportive cells of the inner ear and vestibular organ, but not in the corresponding nerve fibers, which is in contrast to the acutely infected mice. The distribution of the viral genome was further analyzed in adjacent areas of the central nervous system. An involvement of acute and latent herpes virus infection in inner ear dysfunction including sudden deafness and vestibular neuronitis in man, might be suggested from the results described. The presented animal model system, PRV-infected swine, should permit further studies on a possible role of herpetic recurrences, particularly with regard to inner ear disturbances. PMID- 3035931 TI - Cyclophosphamide immunosuppression during lymphotropic herpesvirus infection in the guinea pig model. A histopathologic and virologic study. AB - Guinea pigs infected with lymphotropic herpesvirus (GPHLV) were given the immunosuppressive agent cyclophosphamide (Cy). All Cy-treated animals revealed the expected lymphoid depletion of spleen and lymph node B zones. Acute GPHLV infection of Cy-treated animals resulted in increased blood and spleen leukocyte viral infectivity titers and lymphoid tissue lesions containing cells positive for GPHLV antigen and intranuclear inclusions. During latent GPHLV infection, Cy treatment resulted in declining leukocyte viral infectivity titers without pathologic lesions. Morphologic data suggest that tissue histiocytic cells may be involved in the productive viral infection observed in Cy-immunosuppressed animals during acute GPHLV infection. During latency, however, infectious virus appears restricted to a Cy-sensitive, probably lymphoid, cell. This animal model appears useful for the study of lymphotropic viral infection during immunosuppression. PMID- 3035933 TI - The facial nerve. PMID- 3035935 TI - Measurement of superoxide release from single pulmonary alveolar macrophages. AB - An electrooptical method was developed to quantify superoxide (O2-) release from single rat pulmonary alveolar macrophages (PAM) during adherence to the bottom of a culture dish. This was done by measuring the reduction of nitro blue tetrazolium (NBT) to a diformazan precipitate at 550 nm from videorecorded images of individual cells. Temporal changes in cell optical density, which are proportional to the mass of diformazan produced, were calculated from videophotometric measurements of the change in light intensity over individual cells. Total diformazan produced increased 78 and 126% with an increase in NBT from 0.5 to 1.0 and 2.0 mg/ml, respectively. Total diformazan produced and maximum rate of production among individual PAM varied two- to threefold providing strong evidence for heterogeneity in O2- production. Specific inhibition of O2- production by superoxide dismutase, iodoacetate, and chlorpromazine significantly reduced the total diformazan produced and maximum rate of diformazan production. Hydrogen peroxide was not involved in NBT reduction, since catalase alone did not significantly change diformazan production. This novel method to quantify O2- release from single PAM should be valuable in analyzing heterogeneity and single cell kinetics of O2- production, in assessing the effects of exposure of cells to particulates on O2- release, and in relating release to electrophysiological measurements. PMID- 3035934 TI - Hormone 1 alpha,25-dihydroxyvitamin D3 modulates heat shock response in monocytes. AB - Monocytes, which play a central role in inflammatory reactions, are exposed to elevated temperatures in febrile states or to potential cellular toxins (e.g., reactive oxygen species, immune mediators). We analyzed the heat shock (HS) response in human peripheral blood monocytes and the monocytic line U937 compared with that in dermal fibroblasts. After HS, U937 cells and fibroblasts synthesized mainly the 70- and 83-kDa HS proteins (HSPs), whereas the spectrum of HSPs synthesized by monocytes varied over the range 41-45 degrees C. Incubation with hydrogen peroxide induced synthesis of the 70-kDa HSP in monocytes only. The hormone 1 alpha,25-dihydroxyvitamin D3 [1,25-(OH)2D3] that protects monocytes from thermal injury had similar effects on the HS response in U937 cells, increasing synthesis of HSPs and levels of HSP 70 mRNA and partially preventing the decrease in normal protein synthesis, whereas interferon-gamma had no effect. 1,25-(OH)2D3 did not modify the HS response in fibroblasts, although these cells contained cellular receptors for 1,25-(OH)2D3. 1,25-(OH)2D3 could play a specific role in modulating the HS response in monocytes and related cells. PMID- 3035936 TI - Epinephrine increases plasma immunoreactive atrial natriuretic hormone levels in humans. AB - Six normal human subjects each underwent sequential 80-min infusions of saline and epinephrine (EPI) at 0.55 and 2.75 micrograms X min-1 X m-2 to determine the role of EPI in the control of atrial natriuretic hormone (ANH) in humans. Plasma immunoreactive-ANH (IR-ANH) levels nearly doubled in response to the infusion of EPI at 0.55 microgram X min-1 X m-2 (P less than 0.05) and then plateaued; heart rate accelerated significantly (P less than 0.01) with increasing plasma EPI levels, whereas systolic blood pressure increased only with higher plasma EPI levels (P less than 0.05). To determine whether beta-adrenergic mechanisms mediate the EPI-induced increase in IR-ANH, six additional subjects each received infusions on two separate days of saline for 240 min and the beta-adrenergic antagonist propranolol followed by propranolol plus EPI at 2.75 micrograms X min 1 X min-2 each for 80 min. Neither saline nor propranolol plus EPI caused a significant increase in plasma IR-ANH. We conclude that EPI increases plasma IR ANH through beta-adrenergic mechanisms in humans. beta-Adrenergic-mediated increases in plasma IR-ANH levels appear to be unrelated to changes in the heart rate. PMID- 3035937 TI - Multiple effects of phorbol esters on hormone-sensitive adenylate cyclase activity in S49 lymphoma cells. AB - In S49 lymphoma cells, 12-O-tetradecanoyl phorbol-13-acetate (TPA) enhances adenylate cyclase activity and doubles cAMP accumulation in response to beta adrenergic stimulation at 37 degrees C, putatively via the action of protein kinase C. At 27 degrees C, TPA has the opposite effect, inhibiting cAMP production in response to isoproterenol by approximately 25%. TPA also inhibits the response to prostaglandin E1 (PGE1), another stimulant of hormone-sensitive adenylate cyclase in these cells, by 30% at 37 degrees C and almost 50% at 27 degrees C. In contrast, TPA enhances responses to forskolin and cholera toxin at both 27 and 37 degrees C. In membranes from cells treated with TPA, PGE1 stimulated adenylate cyclase activity is inhibited by 50%, whereas the catalytic activity stimulated by NaF or forskolin is enhanced. TPA reduces the potency of both PGE1 and isoproterenol for cAMP generation by 50%. TPA causes a similar decrease in beta-adrenergic agonist affinity with no reduction in the density of either antagonist or agonist binding sites in wild type cells and in cells lacking the alpha-subunit of the stimulatory transducer protein (Gs) (cyc-) or lacking functional receptor Gs coupling (UNC). Therefore, TPA has at least three functionally distinct effects on hormone-sensitive adenylate cyclase in S49 cells: a 50% reduction in agonist affinity, attenuation of receptor-transducer coupling, and enhancement of GTP-dependent catalytic activity. We conclude that multiple and opposing effects of TPA on hormone-sensitive adenylate cyclase occur simultaneously within the same cell, affecting the responses to several agonists differently.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3035938 TI - Cephalic phase of gastroduodenal alkaline secretion. AB - Alkaline secretion measured under basal conditions in the intact stomach of conscious dogs averaged 47 mumol/30 min and was about twice lower than that recorded in the proximal (approximately 7 cm long) portion of the duodenum. Vagal excitation elicited by sham feeding and insulin resulted in a marked stimulation of alkaline secretion both from the stomach and the duodenum. Atropine significantly reduced gastric and duodenal alkaline secretion under basal state. It abolished gastric and diminished duodenal alkaline response to sham feeding and insulin hypoglycemia, while propranolol was without significant influence. Indomethacin reduced by approximately 75% basal duodenal alkaline secretion but did not prevent the increment in alkaline response to vagal stimulation. We postulate the existence of the cephalic phase of gastroduodenal alkaline secretion, which seems to be cholinergically dependent in the stomach and partly of noncholinergic and nonadrenergic character but prostaglandin dependent in the duodenum. PMID- 3035939 TI - Structural and functional maturation of rat gastrointestinal barrier with thyroxine. AB - It has been noted that the closure of the intestinal barrier to immunoglobulins is a normal maturational process in the rat. It has also been noted that the microvillus membrane (MVM) of newborn animals differs from adult MVM. The purpose of this study is to document whether thyroid hormone can induce closure in vivo in the rat and to relate this effect of thyroxine to the structural and functional maturation of the intestinal MVM. To assess closure, 2-wk-old rats were fed a rat immunoglobulin G (IgG), and serum antibody binding activity was measured 4 h later. The antibody binding activity of treated animals (T) was 1.5 2 times less than that of controls (C) (P less than 0.001), indicating that thyroxine stimulates closure. The MVM similarly showed signs of maturation. Structural maturation was demonstrated by the lower fluidity of the thyroid treated animals' membranes. Under the influence of thyroxine, the number of receptors on the MVM for IgG had decreased [2.8 X 10(-7) M (C) vs. 1.7 X 10(-7) M (T)], while the Ka remained the same, demonstrating the functional maturation of the MVM. In conclusion, thyroid hormone can induce both structural and functional maturation of the intestinal MVM and can enhance the intestinal mucosal barrier by decreasing the penetration of antibodies. PMID- 3035941 TI - Localization of alpha 2-adrenoceptor-mediated increase in renal Na+, K+, and water excretion. AB - Free-flow micropuncture and clearance studies were conducted in male Sprague Dawley rats to investigate the effects of alpha 2-adrenoceptor stimulation on Na+, K+, and water transport along the nephron. Intravenous infusion of the selective alpha 2-adrenoceptor agonist B-HT 933 at 1 mg X kg-1 X h-1 increased urinary flow rate from 16.2 +/- 3.6 to 84.8 +/- 11.9 microliter/min, fractional excretion of Na+ from 1.36 +/- 0.31 to 3.57 +/- 0.52%, and fractional excretion of K+ from 26.9 +/- 3.0 to 42.3 +/- 2.2%, The diuresis, saluresis, and kaliuresis were not the result of increases in glomerular filtration rate or mean arterial blood pressure. Urine osmolality decreased from 1,126 +/- 177 to 325 +/- 33 mosmol/kg water and in 8 of the 11 animals studied B-HT 933 decreased urine osmolality to hyposmotic levels, suggesting a possible interaction between the alpha 2-adrenoceptor agonist and vasopressin. The alpha 2-adrenoceptor antagonist yohimbine (0.25/mg bolus, iv) inhibited the diuresis, saliuresis, and kaliuresis. In micropuncture studies, B-HT 933 was without effect on single-nephron glomerular filtration rate or on Na+, K+, and water transport along the superficial proximal tubule, loop of Henle, or distal tubule. Thus stimulation of alpha 2-adrenoceptors increases Na+, K+, and water excretion by inhibiting tubule reabsorption of these substances at nephron sites beyond the superficial distal tubule, most likely by the collecting tubule. PMID- 3035942 TI - Cytosolic calcium and the action of vasopressin in toad urinary bladder. AB - The effects of experimental procedures believed to increase cytosolic calcium on basal and vasopressin-stimulated osmotic water flow and transepithelial sodium transport were examined in the toad urinary bladder. Exposure of isolated toad bladders to quinidine, calcium ionophores (A23187, X537A), or low-sodium or potassium-free serosal solutions resulted in a dose-dependent decrease in the hydrosmotic response to vasopressin or exogenous adenosine 3',5'-cyclic monophosphate (cAMP). The degree of inhibition of cAMP-induced water flow induced by low-sodium or potassium-free serosal bathing media varied, and in a similar manner, with the serosal calcium concentration. The effects of quinidine sulfate (2 X 10-4 M), X537A (2 X 10(-5) M), and low serosal sodium (20 mM), but not that of A23187 (10(-5) M), were readily reversible. Exposure to quinidine (4 X 10(-4) M), A23187 (10(-5) M), X537A (5 X 10(-6) M), or low serosal sodium (2 mM) also inhibited the basal short-circuit current (SCC). Vasopressin, 4-20 mU/ml, completely overcame the inhibition of the SCC induced by quinidine, A23187, or low serosal sodium, but a submaximal dose of hormone (4 mU/ml) failed to fully reverse the inhibitory effect of X537A, 5 X 10(-6) M. These results are consistent with the view that 1) a Na-Ca exchange process operates across the basolateral surface of the granular epithelial cells of the toad urinary bladder in vivo, and 2) the level of free calcium in the granular cell cytosol plays a modulatory role in the control of apical membrane water and sodium permeability by vasopressin, and in the regulation of the basal rate of transepithelial sodium transport. PMID- 3035940 TI - Absence of a cAMP-mediated antiabsorptive effect in an undifferentiated jejunal epithelium. AB - In the relatively undifferentiated jejunal mucosa occurring in piglet viral enteritis, we measured the response of transepithelial Na+ and Cl- fluxes in vitro to raised intracellular adenosine 3',5'-cyclic monophosphate (cAMP) levels. At the acute 40-h stage of transmissible gastroenteritis (TGE), luminal membrane markers, sucrase and lactase, and a basolateral jejunal epithelial membrane marker Na+-K+-ATPase, were significantly decreased in activity, while a proliferative marker, thymidine kinase, was significantly enriched; these enzyme characteristics are typical of enterocytes isolated from crypts of other species. As expected, control piglet jejunum in short-circuited Ussing chambers after theophylline (10 mM) developed significant net secretory Na and Cl fluxes primarily due to significant antiabsorptive effects (delta JNa m----s = 3.48 +/- 0.52, delta JCl m----s = 2.59 +/- 0.28). Furosemide (10(-4) M), an inhibitor of electroneutral NaCl cotransport, produced antiabsorptive effects (delta JNa m--- s = 2.53 +/- 0.31, delta JCl m----s = 2.58 +/- 0.28) in control jejunum that were not significantly different from those seen in response to theophylline. TGE jejunum, however, responded to theophylline not by an antiabsorptive effect but by significant electrogenic Cl- secretion (delta JCl s----m = 1.59 +/- 0.48); furosemide had no effect on ion fluxes in TGE tissue. Control and TGE jejunal mucosal homogenates did not differ in their basal or theophylline-stimulated levels of cAMP. We conclude that the relatively undifferentiated small intestine occurring in acute TGE does not generate either a cAMP-mediated antiabsorptive effect or a furosemide-mediated antiabsorptive effect.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3035943 TI - Quinidine effect on hydrosmotic response of collecting tubules to vasopressin and cAMP. AB - Quinidine, a compound thought to increase cytosolic calcium ion activity, has been found to inhibit the hydrosmotic response to vasopressin (VP) and adenosine 3',5'-cyclic monophosphate (cAMP) in the toad urinary bladder. To test whether this drug has a similar action in the mammalian nephron, the effect of quinidine on the hydraulic conductivity of the isolated perfused rabbit cortical collecting tubule (CCT) exposed to either 20 microU/ml VP or 10(-4) M 8-(p-chlorophenylthio) - adenosine 3',5' - cyclic monophosphate (8-CPT-cAMP) was studied. Quinidine had no effect on the basal water permeability of the CCT. Quinidine sulfate (10(-4) M) reduced the VP-stimulated water permeability from 280 +/- 50 X 10(-7) to 115 +/- 41 X 10(-7) cm X s-1 X atm-1 (P less than 0.05). The hydrosmotic response to 8-CPTcAMP was likewise reduced following exposure to quinidine. This effect was shown to be dose dependent. In paired experiments, inhibition of the response to 10(-4) M 8-CPTcAMP averaged 11% at 10(-6) M, 27% at 5 X 10(-6) M, 53% at 5 X 10( 5) M, and 50% at 10(-4) M quinidine. Inhibition of the response to 8-CPTcAMP was estimated to be half maximal at approximately 5 X 10(-6) M quinidine. Tubules were protected against the quinidine-induced inhibition by the addition of 6.5 X 10(-5) M quin 2-acetoxymethylester in the presence of low peritubular Ca concentration. These results are consistent with the view that elevated cytosolic Ca ion levels inhibit the increase in water permeability elicited by VP or exogenous cAMP in the mammalian CCT. PMID- 3035945 TI - Sex differences in function and distribution of alpha 1- and alpha 2 adrenoceptors in rabbit urethra. AB - To define the functional role of postsynaptic alpha-adrenoceptors in urethral smooth muscle, we examined the effects of various alpha-adrenergic agonists and antagonists on the contractile properties of the isolated urethra of male and female rabbits and quantified the population of alpha 1-and alpha 2 adrenoceptors, using radioligand receptor binding techniques. Norepinephrine (NE), epinephrine, and phenylephrine, an alpha 1-adrenergic agonist, induced increases in contractile force in both the male and female urethra. Clonidine, an alpha 2-adrenergic agonist, caused a relatively large contractile response in the female urethra but only a small contractile response in the male urethra. Receptor binding studies indicated that the male urethra contains almost equal amounts of alpha 1- and alpha 2-adrenoceptors (32 vs. 34 fmol/mg, respectively), whereas the female urethra contains a significantly greater density of alpha 2- than alpha 1-adrenoceptors (122 vs. 34 fmol/mg, respectively). Our studies indicate that both alpha 1- and alpha 2-adrenoceptors cause contractile responses in male and female rabbit urethra; and the greater response to alpha 2-agonist in female than male urethra is correlated with a higher density of alpha 2 adrenoceptors in this tissue. PMID- 3035946 TI - Myocardial amino acid transport by canine sarcolemma vesicles. AB - The transport of L-alanine and L-leucine into membrane vesicles isolated from mature canine ventricular myocardium was studied. Transport was assessed in purified sarcolemma and in vesicles differentially enriched either for sarcolemma or sarcoplasmic reticulum to further localize these transport systems. An imposed inward gradient of a NaNO3 stimulated uptake of L-alanine but not L-leucine by these vesicles. Amino acid uptake by these vesicles occurred into an osmotically active space. The stimulatory effect of Na+ on alanine transport was most striking in the purified sarcolemma vesicles, where Na+-stimulated alanine flux was 45 +/- 14 pmol X mg-1 X min-1. Furthermore, Na+-dependent alanine transport activity appeared to copurify with Na+-K+-ATPase activity, which served as a marker for sarcolemma membrane when these activities were compared in the three different membrane preparations. Leucine transport by sarcolemma was not altered by an imposed Na+ gradient. However, leucine uptake was a saturable function of extravesicular leucine and was inhibited by valine. In contrast, in sarcoplasmic reticulum membrane vesicles leucine uptake increased proportionately with increasing media leucine and was unaffected by valine. Our results demonstrate the feasibility of directly studying the transport of naturally occurring amino acids in membrane vesicles from mammalian heart, and the presence of Na+ dependent alanine transport system and a Na+-independent leucine transporter in the sarcolemma but not in sarcoplasmic reticulum of canine ventricular myocardium. PMID- 3035944 TI - De novo intrarenal formation of angiotensin II during control and enhanced renin secretion. AB - In previous studies it has not been possible to determine net intrarenal formation of angiotensin II (ANG II) from arteriovenous ANG II concentrations because of the high intrarenal ANG II degradation rates (DR). This study was designed to determine ANG II-DR and to estimate net intrarenal ANG II formation during normal and enhanced renin secretion rate (RSR). In anesthetized dogs, plasma renin activity and ANG II were measured in arterial and renal venous blood by radioimmunoassay during four periods: control, renal arterial constriction (RAC), angiotensin converting enzyme (ACE) inhibition (MK 422), and MK 422 plus systemic arterial ANG II infusion. ANG II-DR was determined in each dog from the arterial-renal venous ANG II concentration difference during the period of ANG II infusion in the presence of ACE inhibition; this value was used to estimate net ANG II formation by predicting the amount of arterially delivered ANG II that escaped degradation. The average percent ANG II-DR calculated during ANG II infusion (range of 0.05 to 0.20 microgram/min) was 89 +/- 2%. In response to RAC, RSR increased from 11 +/- 3 to 24 +/- 5 ng ANG I X h-1 X min-1 X g-1. Arterial ANG II (67 +/- 11 pg/ml) and renal venous ANG II (29 +/- 6 pg/ml) increased to 133 +/- 18 and 61 +/- 10 pg/ml, respectively. Net intrarenal ANG II formation increased from 44 +/- 11 to 83 +/- 13 pg X min-1 X g-1 after renal arterial constriction. There was a significant relationship between the change in RSR and the change in ANG II formation rate.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3035949 TI - Mesenchymal neoplasms in childhood. Insights of Arthur Purdy Stout and where they have taken us. PMID- 3035947 TI - Myocardial adaptation to endurance exercise training in diabetic rats. AB - The purpose of this study was to determine whether exercise training would prevent the progressive functional decline in pump function of hearts from diabetic rats. Four groups were studied: sedentary control, trained control, sedentary diabetic, and trained diabetic. Trained rats were adapted to the treadmill prior to induction of diabetes in half of the group streptozotocin injected (50 mg/kg). Thereafter the duration, speed, and grade were then progressively increased until the trained rats could run for 60 min at 27 m/min, 5% grade (wk 8). Cardiac output and work were measured in isolated working hearts perfused at various left atrial filling pressures and with buffer containing the concentrations of glucose and fatty acids found in vivo. Sedentary diabetic rats had lowered body weight, elevated plasma glucose, triacylglycerol, and cholesterol. Exercise training of diabetic rats lowered plasma triacylglycerol levels. Training increased plantaris muscle cytochrome oxidase activity significantly in both the trained control and trained diabetic groups. Cardiac pump function was impaired in hearts from the sedentary diabetic rats perfused with either normal or diabetic substrate conditions, but the impairment was larger under diabetic conditions. Training of diabetic rats prevented this depression. Myocardial carnitine content was decreased in hearts from sedentary diabetic rats. Exercise training increased carnitine content in both control and diabetic rats. This training protocol did not affect cardiac pump function of the trained control group. These results suggest that exercise training may limit the myocardial contractile dysfunction associated with diabetes mellitus. PMID- 3035948 TI - Alpha-adrenergic receptor subtypes in the cerebral circulation of newborn piglets. AB - The purpose of this study was to identify the alpha-adrenergic receptor subtype mediating cerebral vasoconstriction during sympathetic nerve stimulation in the newborn piglet. The effect of alpha 1- and alpha 2- antagonists prazosin and yohimbine on the cerebrovascular response to unilateral electrical stimulation (15 Hz, 15 V) of the superior cervical sympathetic trunk was studied in 25 newborn piglets. Regional cerebral blood flow was measured with tracer microspheres (15 +/- 1 micron). Sympathetic stimulation decreased blood flow to the ipsilateral cerebrum hippocampus, choroid plexus, and masseter muscle by 15 +/- 2, 10 +/- 2, 51 +/- 5, and 94 +/- 5%, respectively. alpha 1-Adrenergic receptor blockade with prazosin (0.5 mg/kg, n = 10) inhibited the sympathetic vasoconstriction in the cerebrum, hippocampus, and masseter muscle (7 +/- 2, 4 +/ 3, and 55 +/- 9%, respectively) and abolished it in the choroid plexus. alpha 2 Adrenergic receptor blockade with yohimbine (0.5 mg/kg, n = 6 and 1.0 mg/kg, n = 5) had no effect. Following the higher dose of yohimbine, however, blood flow to all brain regions was increased by approximately two-fold, possibly due to enhanced cerebral metabolism. These data demonstrate that vascular alpha 1 adrenergic receptors mediate vasoconstriction to neuroadrenergic stimulation in cerebral resistance vessels in the newborn piglet. PMID- 3035951 TI - Synovial sarcoma. AB - The contributions of Dr. Arthur Purdy Stout to surgical pathology and his observations pertaining to soft tissue tumors (and, more specifically, to synovial sarcoma) are briefly reviewed. In addition, a report on 185 patients treated at the Mayo Clinic for synovial sarcoma is reviewed. In that study, histologic subclassification stratified the tumors as follows: 33% with a predominant biphasic pattern, 31% with a monophasic pattern, and 36% with a mixed pattern. For all 185 patients, the 5-year survival rate was 38% and the 10-year rate was 23%. For patients treated since 1960, the survival rates were 55% at 5 years and 38% at 10 years. Female patients, young patients, and patients with tumors less than 5 cm in diameter had significantly higher survival rates than did their counterparts. Although histologic subtyping did not reveal significant differences in patient survival, patients with glandular differentiation of the epithelial elements seemed to do better. PMID- 3035950 TI - Malignant fibrous histiocytoma 20 years after Stout. PMID- 3035952 TI - Collagenous spherulosis of the breast. AB - Fifteen examples of a hitherto undescribed lesion, which we have designated "collagenous spherulosis," were encountered in breast tissue from women aged 39 to 55 years. The lesion, which was multifocal in eight cases, was an incidental microscopic finding involving lobular acini and ductules, and consisted of intraluminal clusters of eosinophilic spherules measuring approximately 20-100 mu in diameter. The spherules typically were found, and appeared to originate, within the spaces of fenestrated epitheliosis ("papillomatosis"). Special stains indicated that the spherules were collagen-rich, but also contained variable amounts of acidic mucin, PAS-positive, basement membrane-like material, and elastin. With immunoperoxidase staining, the cells immediately surrounding the spherules stained positively for actin, suggesting myoepithelial differentiation. Collagenous spherulosis was typically situated adjacent to, or encompassed by, other benign proliferative processes, including intraductal papillomas, sclerosing adenosis, and "infiltrating epitheliosis" (radial scars). Collagenous spherulosis is a clinically and histologically benign lesion that on microscopic examination has been confused with--and should be distinguished from--malignant lesions including adenoid cystic carcinoma of the breast and so-called intraductal signet-ring carcinoma. PMID- 3035953 TI - Human papillomavirus-associated early vulvar neoplasia investigated by in situ hybridization. AB - Of 21 consecutive cases of early vulvar neoplasia studied at the Istituto Nazionale Tumori of Milan, 62% appeared to be related to papillomavirus infection. This conclusion is the result of the present study by in situ hybridization with DNA probes of human papillomavirus (HPV) 6/11, 16, and 18 and of previous ultrastructural and immunohistochemical investigations. The proportion of cases associated with HPV was 78.5% for those (11/14) with histologic evidence of viral infection and 33% for those without (2/6). HPV 16 was detected in all cases that were positive by in situ hybridization except for one, which showed HPV 6/11 DNA. In one case there was a mixed triple infection for HPV 6/11, 16, and 18. The patient who was positive for HPV 6/11 had a giant condyloma associated with an inguinal lymph node containing a metastatic well differentiated squamous cell carcinoma. Three cases were positive for papillomavirus internal capsid species-nonspecific antigen (PV-Ag) (with ultrastructural evidence of virions in one of them) and were negative for HPV-DNA hybridization. They appeared to be infected with a type of HPV not identified by the available probes. Three cases, and two sites of two other cases with double infection, were HPV-DNA-positive and PV-Ag-negative. They illustrate the limitation of immunohistochemical investigation in cases with high-grade intraepithelial neoplasia. Six cases of verrucous carcinoma of the vulva were negative for HPV DNA by in situ hybridization. PMID- 3035954 TI - The immunohistochemistry of gastrointestinal stromal tumors. Evidence supporting an origin from smooth muscle. AB - To study the histogenesis of gastrointestinal stromal tumors, 79 cases were evaluated for desmin (DES), vimentin (VIM), and S-100 protein immunoreactivity by the avidin-biotin immunoperoxidase procedure on paraffin-embedded, Bouin's-fixed tissue sections. All tumors showed weak vimentin positivity. Trapped non neoplastic smooth muscle and nerve twigs were often noted, particularly at the tumor periphery. Significant tumor S-100 positivity was not identified in our series. Similarly, glial fibrillary acidic protein (GFAP) immunoreactivity (performed in 11 desmin-negative tumors) was not detected within either gastrointestinal stromal tumors or enteric glial cells. Fifty-three percent (53%) of all gastrointestinal stromal tumors (GIST) displayed positive tumor cell desmin immunoreactivity. All 10 esophageal and all four colorectal tumors were diffusely desmin positive and unequivocal smooth muscle lesions. In contrast, only 17 of 37 (46%) benign and six of nine (67%) malignant gastric tumors were desmin positive. Similarly, four of 10 (40%) benign and one of nine (11%) malignant small-bowel tumors expressed desmin. Several gastric neoplasms with prominent nuclear palisading resembling schwannian Antoni A regions were nonetheless desmin positive. Epithelioid gastric tumors were more frequently desmin positive than nonepithelioid tumors; however, this positivity did not attain statistical significance. Two gastrointestinal stromal tumors that were desmin negative in paraffin-embedded material had detectable antigen in frozen sections. Gastric and small-bowel tumors measuring less than 3 cm were significantly more often desmin positive than those 3 cm or greater. We conclude that the method of fixation, tissue preparation, and immunostaining may significantly affect the expression of desmin. Although the histogenesis of gastrointestinal stromal tumors remains controversial, most of these tumors show evidence of smooth muscle differentiation. PMID- 3035955 TI - Invasive benign histiocytoma. PMID- 3035957 TI - Congenital and neonatal malignant tumors. A 28-year experience at Children's Hospital of Los Angeles. AB - Fifty-one cases of congenital and neonatal malignant tumors were collected from the Children's Hospital of Los Angeles Department of Pathology files and reviewed. The study covered a 28-year period, 1958-1985. Thirty (59%) of the patients died. The types, incidence, clinical features, and behavior of neoplasms occurring in the neonate were different from those seen in older children and adolescents. Moreover, the response to therapy was also dissimilar. Leukemia and neuroblastoma were the most frequent malignancies and were responsible also for the largest number of deaths. PMID- 3035956 TI - Superoxide generation of leukemic cells in children. AB - Superoxide (O2-) generation of leukemic cells was investigated in 28 children with acute leukemia. Leukemic cells of seven with acute myelomonocytic leukemia and one with acute monocytic leukemia produced various amounts of O2- (0.13-0.87 nmol/min/10(6) cells) when stimulated with wheat germ agglutinin (WGA) and phorbol myristate acetate (PMA). Other types of leukemia cells exhibited no such functions. Leukemic cells of two patients diagnosed as unusual cases of acute lymphoblastic leukemia generated significant amounts of O2- when stimulated with WGA and PMA. Both patients went into complete remission while on acute nonlymphoblastic leukemia treatment. Thus O2- generating activity can serve as a functional marker of leukemic cells in myelomonocytic lineage. PMID- 3035958 TI - Immunoregulatory T cells in males vulnerable to Epstein-Barr virus with the X linked lymphoproliferative syndrome. AB - The X-linked lymphoproliferative syndrome (XLP) is a primary immune deficiency. Affected males are vulnerable to fatal infection by Epstein-Barr virus (EBV). In patients who had survived an infection by EBV, helper and cytotoxic T cells reactive to virus-transformed autologous B lymphoblastoid cells were infrequent. This was similar to that observed in normal individuals seronegative to EBV. In contrast, frequencies of the reactive T cells were high in normal controls seropositive to EBV. Patients with XLP also had a propensity to activate radiosensitive suppressor cells when their T cells were stimulated by EBV transformed B cells. Hence, clonal expansion of helper and cytotoxic T cells to virus-transformed B cells failed to occur during an EBV infection in these patients because of immunosuppression. These defects in the T cell repertoire of patients with XLP may predispose them to be susceptible to EBV-induced lymphoproliferation and contribute to the variability in expression of the phenotypes seen in these patients. PMID- 3035959 TI - Drug use among a sample of males admitted to an alcohol detoxication center. AB - Urine samples were obtained from 93% of a sample of 111 consecutive male admissions to a nonmedical detoxication center in Toronto. Analysis of these samples revealed that 51 (50%) had traces of drugs other than alcohol and that 12 (12%) were alcohol free. Benzodiazepines were the most frequently detected drugs and these were found in 32% of all samples. Barbiturates were detected in seven (7%) samples and cannabinoids in 10 (10%) samples. A wide range of urine alcohol concentrations was found and some samples had zero or low concentrations. Although alcohol urine concentrations were generally lower in samples containing other drugs, the distribution of urine alcohol concentrations was similar for samples containing only alcohol and those containing alcohol and some other drug. Of the 51 samples found to contain drugs other than alcohol, subjects' self reports were concordant in 27 cases (53%). Most of the discrepancies between self reports and urine analysis were due to the under-reporting of the use of benzodiazepines. Subjects with drugs in their urine tended to be younger than others, but they were not distinguished with respect to their behaviors while in the detoxication center or length of stay. Those with the highest urine alcohol concentrations had shorter stays in the detoxication center. Implications for further studies are discussed. PMID- 3035960 TI - Seasonal allergic rhinitis treated with a beta-2-adrenostimulant. AB - The aim of the study was to investigate whether topical application of a beta-2 adrenostimulant (fenoterol) on the mucous membrane has a clinically significant anti-allergic effect. Thirty-three patients with grass pollen hay fever completed the trial which was a double-blind, placebo-controlled cross-over design. After a run-in period the patients received two puffs of 50 micrograms fenoterol 4 times a day or placebo for 21/2 weeks before cross-over. Symptoms were scored on diary cards and there was a moderate, but significant effect of fenoterol on sneezing, but the effect on secretion and blockage was insignificant. It is concluded that beta-2-stimulating agents are not competitors to the very effective topical steroids. PMID- 3035961 TI - [Adenosine triphosphatases of fetal large intestine epithelium. Light and electron microscopy studies of the cecum of cattle (Bos primigenius taurus)]. PMID- 3035962 TI - Morphologic and histochemical analysis of the newt (Notophthalmus viridescens) liver. AB - Architectural arrangement, ultrastructure, and selected histochemical properties of the newt (Notophthalmus viridescens) liver were examined. Although hematopoietic tissue (1-4 cells thick) invested the liver, direct vascular communication between this tissue and hepatic parenchyma was not observed. The liver was intensely positive when stained with Oil-red-O and periodic acid-Schiff reagent and connective tissue was limited to large vascular channels and the capsule. A distinctive polarity was observed in the hepatic vascular system when lobes were viewed in cross section. Dorsally, portal venules accompanied arterioles and branches of the biliary system, while tributaries of hepatic veins were observed ventrally. Following perfusion fixation, hepatocytes appeared as sheets of cells 1-5 cells thick; however, lobules as defined in adult mammalian liver were absent. Hepatocytes contained abundant smooth endoplasmic reticulum, mitochondria, electron-dense lysosomes, patches of granular endoplasmic reticulum, and lipid droplets. Continuous endothelial cells lined sinusoids and exhibited fenestrae organized into structures similar to sieve plates observed in mammalian liver. Variable numbers of melanin-containing macrophages and subendothelial macrophages were observed; however, Kupffer cells and lipid containing perisinusoidal fat-storing cells were not seen. Patterns of reaction product for glucose-6-phosphatase (G-6-Pase), glucose-6-phosphate dehydrogenase (G-6-PDH), and succinic dehydrogenase (SDH) were localized in the newt liver. All enzymes exhibited a uniform distribution pattern; however, small punctate regions of intensely positive G-6-PDH cells were noted within hepatic parenchyma. Cells comprising the hematopoietic tissue were intensely positive for G-6-Pase, G-6 PHD, and negative for SDH. PMID- 3035963 TI - The effect of propofol on adrenocortical steroidogenesis: a comparative study with etomidate and thiopental. PMID- 3035964 TI - Effect of angiotensin blockade and converting enzyme inhibition on renovascular hypertension: comparison between unilateral and bilateral renal artery stenosis. AB - The response to angiotensin II analog infusion during sodium deletion and the effects of a one-month captopril treatment were compared between 15 renovascular hypertensive patients with unilateral and 6 with bilateral renal artery stenosis. Plasma renin activity, its response to sodium depletion, and the renal vein renin ratio during sodium depletion were greater in unilateral than in bilateral stenosis. A fall in diastolic blood pressure induced by analog infusion during sodium depletion was correlated with the preinfusion plasma renin activity and with the renal vein renin ratio. Treatment with captopril showed a comparable hypotensive effect in unilateral and bilateral stenosis. The reduction in blood pressure was not correlated with the pretreatment renin levels or changes in blood pressure observed during analog infusion. Plasma renin activity rose and plasma aldosterone level fell in all patients. These results indicate that the mechanism maintaining high blood pressure is more renin dependent in unilateral than in bilateral stenosis and that the long-term effect of captopril does not depend solely on the suppression of the renin-angiotensin-aldosterone system. PMID- 3035965 TI - Estimation of endothelial receptor sites with the mean transit time approach. AB - A new procedure for quantitating endothelial receptor sites is described. The mean transit time of the passage of an appropriate radio-labeled ligand from the artery to vein of an organ is compared to the mean transit times of a vascular indicator and a water label. The concentration of the unlabeled ligand is progressively increased to define that concentration at which half of the enzyme sites are occupied. Values are calculated for the number of receptor sites accessible during a single circulation in each ml of exchangeable tissue water. This approach is illustrated by estimating the sites of carbonic anhydrase present on the pulmonary endothelium, utilizing labeled acetazolamide as a ligand. Preliminary studies of the receptor sites on isolated endothelial cells suspended in an elutriator are also presented. PMID- 3035966 TI - Effects of changes in pulmonary perfusion and surface area on endothelial ACE activity. AB - Measurement of transport or enzymatic processes associated with pulmonary endothelial cells could provide unique information regarding the physiologic function of the microcirculation as well as data describing the biochemical integrity of these cells in acute lung injury. Since an important goal of such measurements is to quantify the kinetics of reactions of substrates (indicators) with the endothelium, it would be highly advantageous to account for (convective) phenomena related to blood flow and its distribution which also influence whole organ metabolism. In the present report, we describe studies that have utilized a nonlinear model of organ metabolism to estimate Vmax, the apparent maximal velocity and Km, concentration at one-half Vmax of angiotensin-converting enzyme (ACE) activity. Our hypotheses have been that (a) there is sufficient data to calculate apparent kinetic constants from indicator-dilution outflow curves after injection into the pulmonary circulation of a radiolabelled synthetic substrate for ACE, and (b) Vmax for ACE is a property related to the amount of enzyme located on the endothelial cells, and as such, should be directly proportional to perfused surface area of an organ. In isolated perfused rabbit lungs, when flow was increased over a two-fold range, but surface area was unchanged, neither Vmax nor Km for ACE activity was significantly altered. When indicator-dilution measurements of pulmonary ACE activity were made in lambs from 1-171 days of age there was a progressive increase in Vmax with no significant change in Km as a function of age. The increase in Vmax was closely correlated with an independent measure of surface area, carbon monoxide diffusing capacity, and a morphometric measure of capillary endothelial cell surface area (stereology at electron microscopic level) made at post-mortem. These experimental observations in whole organs support our hypotheses and predictions regarding the metabolism or removal of substrates from the circulation by means of a saturable process. PMID- 3035967 TI - The effect of intraosseous sodium bicarbonate on bone in swine. AB - Five domestic swine weighing 8 to 12 kg were anesthetized with ketamine 20 mg/kg IM and pentobarbital 20 mg/kg IV. After a sterile prep each animal received NaHCO3 1 mEq/mL/kg in one tibia and saline 1 mL/kg in the other. The animals were allowed to recover and were observed for 30 days. At the end of this period, roentgenographs were obtained of each tibia and triple phase 99m technetium bone scans were obtained. The tibias also were sectioned, stained, and examined under light microscopy for microscopic abnormalities. The only deficit found was a small cortical calcification at the site of the needle puncture in an animal that received NaHCO3. All other roentgenographs, bone scans, and microscopic specimens were normal. This study demonstrates that NaHCO3 does not have permanent adverse effects when injected into the marrow cavity of swine and supports previous clinical observations regarding the safety of NaHCO3. PMID- 3035968 TI - Importance of the medullary macrophage in the replication of lymphoid leukosis virus in the bursa of Fabricius of chickens. AB - Electron microscopy and immunocytochemistry were used to study the development of lymphoid leukosis virus infection in the bursa of Fabricius of experimentally infected chicken embryos and chickens. In embryos infected at 7 days of incubation and killed 10 days later, virus particles and group-specific viral antigen were confined mainly to the connective tissue of the lamina propria of the bursal mucosal folds; a few developing follicles had discrete virions and group-specific antigen between cells. In chickens infected at 1 day of age, infection (as determined by use of electron microscopy and immunocytochemistry) was maximal in 1- to 4-month-old birds, and the greatest concentration of virus and group-specific viral antigen was in the medulla of the follicles. Although lymphoid leukosis virus was released from lymphocytes, epithelial cells, and macrophages, virus replication in the medullary macrophages was more active than that in the other cells. Normal medullary macrophages had cell membrane vesicles (50 to 80 nm in diameter) that covered part of all of the cell membrane surface. In infected chickens, virus particles frequently developed within these vesicles. Comparable vesicles were not found on cortical macrophages. Results of the present study indicated that the medullary macrophage was the principal host cell for replication of lymphoid leukosis virus in the bursa of Fabricius of the chicken. PMID- 3035969 TI - Hormonal changes in sows after induced porcine parvovirus infection in early pregnancy. AB - Hormonal changes, lesions, and virus isolation studies were determined in sows after uterine artery inoculation with porcine parvovirus [( PPV], strain NADL-8) in early pregnancy. Two sows were given PPV on days 14 or 16 and were euthanatized and necropsied on day 35 after twice daily plasma collection for hormone measurement. Parvovirus was given to 4 sows on day 14 and to 4 sows on day 21 with 5 times daily plasma samples collected for 1 week. Sows were examined on days 21 and 28, respectively. Four control sows in each group on days 14 and 21 were given a placebo injection and were similarly studied. All embryos in all but 1 sow given PPV were in various stages of resorption at necropsy. Normal embryos were present in all control sows. Estrone sulfate values increased logarithmically, progesterone values remained stable, and concentrations of 13, 14-dihydro-15-keto-prostaglandin (PG) F2 alpha (PGFM), a PGF2 alpha metabolite, were less than 200 ng/ml for sows given a placebo. In contrast, sows with resorbing embryos did not have an increase in estrone sulfate values. A decrease in plasma progesterone values occurred in 9 of 10 sows inoculated with PPV; this decrease was accompanied by greater than or equal to 1 marked increase in PGFM concentrations. Quantitative assessment of the uterus revealed significantly greater cytoplasmic density in endometrial and glandular cell (P less than 0.01), a greater glandular epithelium height (P less than 0.05), and twice the number of glands (P less than 0.05) in control sows, compared with values in sows inoculated with PPV.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3035970 TI - Rectal transmission of bovine leukemia virus in cattle and sheep. AB - Bovine leukemia virus (BLV) was transmitted by rectal inoculation of BLV infective whole blood into cattle and sheep. Two cows and 2 sheep each were given 500 ml and 50 ml of blood, respectively, by rectal infusion. Two sheep which served as positive controls each were given 1 ml of the same blood, IV. All animals became seropositive to BLV by postinoculation week 5. Although relatively large volumes of blood were used for rectal inoculation, a base line for infectivity was established for the rectal route. PMID- 3035971 TI - Clinical, virologic, and histopathologic observations of induced porcine parvovirus infection in boars. AB - Twelve 8- to 12-month-old crossbred boars were inoculated with a virulent strain (NADL-8) of porcine parvovirus (PPV). Hemicastrations were performed on 6 boars 3, 7, 10, 14, 21, and 28 days after an IM injection of 10(8) median cell culture infectious dose (CCID50) of PPV (n = 3) or injection of 10(7.4) CCID50 given intratesticularly (IT, n = 3). Noninfected cell culture medium (0.25 ml) was injected into each testicle of a 7th boar (IT inoculated control). Virus or viral antigen was detected in testicular and epididymal tissues up to 14 days after inoculation. Direct immunofluorescence indicated that viral antigen was mainly associated with the vasculature of the interstitium. Microscopic lesions were not evident in the testicles and epididymides of IM inoculated boars. Acute-to chronic testicular degeneration was evident in the IT inoculated boars, as well as in the IT inoculated control boar. Six boars were inoculated IM or orally/nasally with 10(7.9) CCID50 of PPV. Semen was collected twice weekly for 8 weeks after inoculation. Virus was not detected in any ejaculates. Semen also was collected from 4 boars for 5 weeks before inoculation, and preinoculation and post-inoculation semen quality was compared. Pronounced changes in sperm output, ejaculate volume, motility, or morphologic defects were not observed. The reproductive consequences of experimental PPV infection in boars were minimal because reproductive function was unaffected and venereal transmission of PPV was not detected. PMID- 3035973 TI - Parvovirus-induced regression of canine transmissible venereal sarcoma. AB - In 1981, during the worldwide canine parvovirus (PV) epizootic, canine transmissible venereal sarcomas growing in Beagles in a colony regressed earlier than expected after the dogs became infected with PV. Subsequent studies revealed that modified-live PV vaccine (feline panleukopenia virus) was capable of preventing tumor transplantation when the vaccine was inoculated simultaneously with the tumor in a site distant from the implantation site. However, the PV vaccine had no effect if it was inoculated 3 or 18 days after the tumor was transplanted. PMID- 3035972 TI - Second-generation pseudorabies virus vaccine with deletions in thymidine kinase and glycoprotein genes. AB - A modified-live pseudorabies virus (PRV) vaccine, designated PRV(dlg92/d1tk), with deletions in the thymidine kinase (tk) and glycoprotein-gIII (g92) genes, was derived from the PRV (Bucharest [BUK]-d13) vaccine strain. The vaccine virus also contained a deletion in glycoprotein gI. Despite 3 deletions, PRV(dlg92/d1tk) replicated to high titers in cell culture from 30 C to 39.1 C. Enzyme assays and autoradiography revealed that PRV(dlg92/d1tk) did not induce a functional tk activity in infected tk- RAB(BU) cells (rabbit skin). Rabbit skin cells were infected with PRV(dlg92/d1tk), with vaccine strains derived from BUK or Bartha K strains of PRV or with the virulent Illinois (ILL), Indiana Funkhauser (IND-F), and Aujeszky (Auj) strains of PRV and were labeled with [3H]mannose from 4 or 5 to 24 hours after infection to investigate whether these viruses induced the synthesis of glycoprotein gIII. Nonionic detergent extracts were prepared and immunoprecipitated with antisera from pigs vaccinated with tk( )-PRV(BUK-d13) or tk+-Bartha K, pigs vaccinated with tk+-PRV(BUK) strains and then challenge exposed to tk+-PRV(IND-F), naturally infected domestic or feral pigs, and pigs vaccinated with tk-)-PRV(dlg92/d1tk). Mouse monoclonal antibodies against PRV glycoproteins gIII, gp50, and gII were also studied. After immunoprecipitation, labeled PRV-specific proteins were analyzed by sodium dodecylsulfate-polyacrylamide gel electrophoresis and autoradiography. The PRV glycoprotein-gII complex, but not glycoprotein gIII, was synthesized in PRV(dlg92/d1tk)-infected cells. Glycoprotein gII and gIII were made in cells infected with PRV vaccine strains BUK, Bartha K, and BUK-d13 and with virulent PRV strains ILL, IND-F, and Auj. Cells infected with PRV(dlg92/d1tk) and with PRV strains ILL, IND-F, Auj, Bartha K, BUK, and BUK-d13, excreted into the cell culture medium a highly sulfated glycoprotein gX of about 90 kilodaltons. Antibodies to glycoprotein gIII were not detected in the sera of pigs inoculated with PRV(dlg92/d1tk), but were found in all other swine sera. PMID- 3035974 TI - A survey of the respiratory health of silica-exposed gemstone workers in Hong Kong. AB - Respiratory symptoms and radiographic and lung function alterations were studied in a cross-sectional survey of gemstone workers in Hong Kong. The study population included a group of grinders, polishers, and buffers who were heavily exposed to dust (principally free silica) and a less exposed group of cutters and carvers. Among all 218 male workers who answered the respiratory questionnaire, heavily exposed workers reported significantly higher prevalence of respiratory symptoms suggestive of mucus hypersecretion. Radiological pneumoconiosis defined as opacities with profusion of 1/0 and above was found in 27% of 157 workers who accepted radiographic and lung function examination. Radiological opacities were significantly related to increasing years of employment in both groups of workers after taking into account age and smoking habits. Decline in forced vital capacity (FVC) was significantly related to increasing years of employment in both groups after allowing for the effects of age, height, and smoking. A modest decline in forced expiratory volume in one second (FEV1) was related to dust exposure which was of borderline statistical significance in polishers and buffers who smoked. Radiological pneumoconiosis did not have an independent effect on lung function when allowance was made for dust exposure. PMID- 3035975 TI - Direct effects of neutrophil oxidants on elastase-induced extracellular matrix proteolysis. AB - Oxidant species produced by human polymorphonuclear leukocytes (PMN) inactivate alpha-1-protease inhibitor and thus may indirectly enhance neutrophil elastase induced proteolysis. It is unclear, however, if PMN-derived oxidants directly enhance proteolysis of extracellular matrix by neutrophil elastase. Matrix was produced by neonatal rat aortic smooth muscle cells and pulse-labeled with 3H lysine to allow identification of the collagen-specific amino acid, hydroxylysine (3H-HL), and the elastin specific amino acid, desmosine (3H-DES). The smooth muscle cells were lysed, and the remaining matrix was used as a culture surface and a proteolytic substrate for intact PMN and purified neutrophil elastase. Proteolysis of collagen and elastin were quantified by chromatographic separation of the marker amino acids 3H-HL and 3H-DES, which were released into the supernatant or remained in the matrix after a 3-h incubation at 37 degrees C. The peptide, formyl-methionine-leucine-phenylalanine (FMLP), produced more rapid release of myeloperoxidase than did phorbol myristate acetate (PMA), which produced more release of O2- and H2O2 than did FMLP. The percent release of total matrix 3H-DES in the presence of PMN + FMLP was 2.45 +/- 0.19% (mean +/- SE, n = 6) and with PMN + PMA it was 1.32 +/- 0.1% (n = 6, p less than 0.01). The release of matrix 3H-HL did not differ. Neutrophil cytoplasts, which produced O-2 and H2O2 but lacked azurophilic granules, did not significantly enhance either elastin or collagen degradation by purified neutrophil elastase (NE).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3035976 TI - Pulmonary alveolar macrophage function during acute inflammatory lung injury. AB - Pulmonary alveolar macrophages (PAM) are present during acute lung inflammation, yet the functional role of these cells in both the initiation and resolution of lung injury is not well defined. To better understand the relationship between PAM functional responses and the evolution of acute reversible lung injury, we examined the ability of both unstimulated and stimulated (PMA, zymosan) PAM to secrete reactive oxygen metabolites (superoxide anion O2-) and lysosomal enzymes (lysozyme, N-acetyl-B-D-glucosaminidase) at specific time points (0, 6, 12, 24, 48, and 72 h) after initiation of acute lung injury via reverse passive Arthus reaction in pathogen-free Sprague-Dawley rats. After acute lung injury, stimulated PAM produced increasing amounts of O2- compared with PAM from noninjured lungs. Maximal O2- production by PAM occurred at 24 h after lung injury, at which time a 3.5-fold and 50% increase in O2- production by PAM was observed when PAM were stimulated with PMA and zymosan, respectively. The amount of O2- generated by these cells slowly decreased during the next 48 h. Enhanced generation of O2- by PAM from injured lungs was not due to altered enzymatic activity of the O2--producing NADPH oxidase, nor was it due to an absolute increase in the NADPH oxidase in "activated" PAM. These observations suggest that increased O2- generation by PAM from injured lungs is due to enhancement of mechanisms responsible for induction of oxidase activity. In addition, a differential accumulation and secretion of lysozyme and N-acetyl-B-D glucosaminidase activity by PAM was observed after acute lung injury.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3035977 TI - Degradation of collagen in lung tissue slices exposed to hyperoxia. AB - Exposure of animals to oxidant gases produces a mild emphysema, and O2-derived free radicals are capable of degrading connective tissues in vitro. It is postulated that degradation of connective tissue by O2-derived free radicals leads to emphysema in these models. To determine whether exposure of lung tissue slices to an oxidant gas results in degradation of collagen and to investigate factors mediating this degradation, we exposed lung tissue slices from normal rats to hyperoxia (95% O2, 5% CO2) and measured hydroxyproline release into the medium. After a 4-h exposure, the hydroxyproline released was 5.3 +/- 0.2 micrograms/g lung tissue (n = 10) in normoxia and 8.1 +/- 0.6 micrograms/g tissue (n = 13) in hyperoxia (p less than 0.05), suggesting degradation of collagen. The addition of 0.1% trypsin to the initial incubation medium caused a synergistic increase in hydroxyproline release from O2-exposed slices: normoxia/trypsin, 46.2 +/- 3.6 micrograms/g tissue (n = 10); hyperoxia/trypsin, 61.4 +/- 3.6 micrograms/g tissue (n = 11) (p less than 0.05). The addition of proteinase inhibitors completely suppressed the O2-induced release of hydroxyproline, suggesting that proteolytic enzymes are involved in hyperoxia-mediated degradation of lung collagen. PMID- 3035978 TI - Is leukotriene B4 an important mediator in human IgE-mediated allergic reactions? PMID- 3035979 TI - Pancreatitis associated with isotretinoin-induced hypertriglyceridemia. PMID- 3035980 TI - [A case of acute sensory and autonomic neuropathy with regression]. AB - The authors report the case of a 24 year old man with no previous disease who presented with a severe autonomic neuropathy. This included major gastrointestinal dysfunction characterised by decreased peristalsis without distension and paralysis of the gall bladder, and orthostatic hypotension with a normal cardiac tachycardia reflex. There was an associated sensory neuropathy affecting heat sensitivity without motor dysfunction and an increased CSF protein content. The proprioceptive nerve fibre conduction was decreased but another nerve conduction was normal initially. The mesenteric plexuses examined during sigmoidectomy performed for peritonitis due to multiple bowel perforations caused by fecoliths, showed no significant changes. Peripheral nerve biopsy revealed massive rarefaction of myelinated fibres which were of small diameter, and of the unmyelinated fibres, mainly due to axonal degeneration. Only two similar cases with incomplete recovery were found in the literature. In our case, a complete recovery was observed. The cause of the condition is unknown. PMID- 3035981 TI - [Endocrine cancers of the pancreas]. PMID- 3035982 TI - [Inappropriate secretion of ADH in neuromeningeal sarcoidosis]. PMID- 3035984 TI - Regulation of enkephalin, VIP, and chromogranin biosynthesis in actively secreting chromaffin cells. Multiple strategies for multiple peptides. AB - Enkephalins, vasoactive intestinal polypeptide, and chromogranin A are all contained in the secretory vesicles of chromaffin cells in culture, and are all released from this compartment by secretagogues in a calcium-dependent way. The biosynthesis of each of these peptides, however, is under quite independent regulation. The synthesis and secretion of enkephalin is tightly coupled to acetylcholine and elevated potassium stimulation by calcium influx. Once calcium enters the cell, calcium acts at pharmacologically distinct sites to elicit secretion and enhanced biosynthesis of Metenkephalin. This is demonstrated by the calcium-independent stimulation of enkephalin secretion by 1 mM barium, in contrast to the dependence on extracellular calcium of barium-stimulated biosynthesis of this peptide. The synthesis and secretion of VIP is also coupled to acetylcholine and elevated potassium stimulation by calcium influx. Treatment with barium demonstrates that calcium acts at distinct sites to stimulate secretion and biosynthesis of this peptide; however induction of VIP by barium and veratridine shows greater sensitivity to the calcium channel blocker methoxyverapamil (D600) than does the induction of Met-enkephalin by these agents. These differences in D600 sensitivity may be due to differences in calcium metabolism or voltage-dependent calcium channels in enkephalin-producing and VIP-inducible subpopulations of chromaffin cells. Chromogranin A levels are essentially unaffected by any of the agents which increase enkephalin and VIP levels, although it is secreted in parallel with enkephalins and catecholamines from chromaffin cells in response to secretagogues. We suggest that peptide hormones such as VIP and enkephalins are regulated by calcium-dependent stimulus secretion-synthesis coupling in the chromaffin cell. Cyclic AMP is a positive regulator of enkephalin and VIP biosynthesis, but does not affect acute release of these peptides. The cAMP/protein kinase A system may be a distal mediator of peptide biosynthesis stimulated by secretagogues. Alternatively, cAMP may be involved in early developmental establishment of phenotype or long-term regulation of peptide biosynthesis by other hormones or neurotransmitters. Chromogranin A may represent a class of intravesicular, soluble proteins that are expressed constitutively by the chromaffin cell in the presence or absence of positive regulators of other systems. The biosynthesis of chromogranin A may be coupled to the production or assembly of the secretory vesicle itself. PMID- 3035983 TI - Acetylcholine transport: fundamental properties and effects of pharmacologic agents. PMID- 3035985 TI - Tubulin- and actin-binding proteins in chromaffin cells. PMID- 3035986 TI - Factors controlling packaging of peptide hormones into secretory granules. AB - Endocrine, exocrine, and neuronal cells package only a subset of their secretory products into the electron-dense secretory granules. To investigate the factors controlling selective packaging of proteins into these granules, we utilized the mouse pituitary tumor cell line, AtT-20, which retained the capability to sort adrenocorticotropic hormone (ACTH) into secretory granules in vitro. Packaging of ACTH was blocked by treatment with weak bases, but was unaffected when N-linked glycosylation or sulfation was inhibited. To test whether the targeting information is specified by sorting domains present on peptide hormone sequences, we determined if a protein could be diverted to the dense secretory granules by attachment to a peptide hormone sequence. A plasmid DNA was constructed that encoded a hybrid protein in which a fragment of a viral membrane protein was fused to the carboxy terminus of human growth hormone. AtT-20 cells transfected with the hybrid were found to target it to dense secretory vesicles efficiently. These results support the hypothesis that sorting domains on peptide hormones direct their packaging into dense secretory vesicles. PMID- 3035988 TI - An integrated approach to secretion. Phosphorylation and Ca2+-dependent binding of proteins associated with chromaffin granules. AB - Pharmacological and morphological studies of secretion from bovine chromaffin cells indicate that the nicotinic receptor initiates intracellular signaling. An increase in [Ca2+]i is a necessary but not sufficient element for secretion. Receptor-dependent, but intracellular-Ca2+-independent alterations of the organization of cortical zone allows close approach of chromaffin granules and plasma membrane. Chromaffin granules possess both Ca2+-dependent and Ca2+ independent protein kinases, but the major substrates for these kinases appear to be associated with other organelles or to be soluble cytosolic proteins. Quantitatively the most important Ca2+-dependent cytosolic components that interact with the chromaffin granule do not show strict specificity for this secretory organelle, but are widely distributed in different cell types and are localized at or close to the plasma membrane in intact chromaffin and other cells. These molecules are closely related in biochemical properties and sequence and include substrates for membrane-associated kinases. Two of these proteins (caldesmon and p36) have binding sites for F-actin; the others described have binding sites for acidic phospholipids. PMID- 3035987 TI - Are changes in intracellular free calcium necessary for regulating secretion in parathyroid cells? PMID- 3035989 TI - [Microradiographic analysis of bone opacities of metastatic origin]. PMID- 3035990 TI - [Gastric metastases of breast cancer. A case of linitis plastica type]. PMID- 3035991 TI - [Intraoperative echography: an indispensable element in surgery of small hepatocarcinomas in cirrhotic patients]. PMID- 3035992 TI - Comparison of horseradish peroxidase and alkaline phosphatase-labelled antibodies in enzyme immunoassays. AB - The periodate method was found to be most effective for preparing horseradish peroxidase-sheep anti-human and horseradish peroxidase-donkey anti-mouse immunoglobulin (IgG) conjugates. The conjugates were improved by carrying out the oxidation of the enzyme at low pH. Anti-human and anti-mouse IgG-peroxidase conjugates (0.5 mg/mL IgG and 0.7 mg/mL IgG, respectively) were used at 1:15,000 and 1:8000 dilutions, respectively, in a sandwich ELISA to detect human and mouse IgG in buffer or in a growth medium containing 20% foetal calf serum. Using the peroxidase conjugates, it was possible to detect human and mouse IgG at concentrations as low as 1 ng/mL. The glutaraldehyde method was found to be much more effective than the periodate method for conjugating alkaline phosphatase to the antibodies. The optimum dilutions for anti/human and anti-mouse IgG-alkaline phosphatase conjugates (0.18 mg/mL IgG and 0.3 mg/mL IgG, respectively) in ELISA were 1:500 and 1:1000, respectively. The detection limit with alkaline phosphatase conjugates was 7 ng/ml for human IgG and 4 ng/ml for mouse IgG. PMID- 3035994 TI - Dietary fibre and age at menarche: a re-examination of the evidence. PMID- 3035993 TI - Disability, rehabilitation and after-care of stroke patients after discharge from hospital, Singapore 1983-84. AB - 135 new onset stroke patients admitted over a nine-month period to a medical unit in Singapore were studied with emphasis on their rehabilitation and community care after discharge. Follow-up visits were made to 79 survivors at third month after onset. 16.7% of the patients made apparently complete recoveries, 20.3% remained slightly disabled, 21.5% moderately disabled, and the remaining 41.8% severely disabled. Sixty-five patients were staying in private households at third month follow-up. The main care-givers were usually female relatives. These informal carers assisted a large proportion of patients in various activities of living: 62.0% in dressing, 54.4% in walking and toileting, 30.4% in feeding, and 22.8% in turning in bed. The study establishes the need to develop and strengthen supportive services to enable disabled stroke patients to be cared for in their own homes. These include the home nursing service, elderly day care service, home help service, and meals-on-wheel service. PMID- 3035995 TI - Chronic food restriction and the circadian rhythms of pituitary-adrenal hormones, growth hormone and thyroid-stimulating hormone. AB - Adult male Sprague-Dawley rats were subjected to food restriction so that they ate 65% of food ingested by control rats. While control rats had free access to food over the 24-hour period, food-restricted rats were provided with food daily at 10 a.m. The experimental period lasted for 34 days. On day 35, rats from both experimental groups were killed at 08.00, 11.00, 14.00, 24.00 and 02.00 h. Food restriction modified the circadian rhythms of ACTH and corticosterone. In addition, total circulating corticosterone throughout the day was higher in food restricted than in control rats. In contrast, food restriction resulted in depressed secretion of thyroid-stimulating hormone and growth hormone. The results indicate that time of food availability entrained circadian corticosterone rhythm but not thyroid-stimulating hormone and growth hormone rhythms. PMID- 3035996 TI - The regulation of cyclic AMP levels in cultured MH1C1 rat hepatoma cells and in solid tumours derived from MH1C1 cell inoculates. AB - Several studies have found high cAMP content in hepatomas in vivo, while hepatoma cells in vitro have very low levels. To explore this discrepancy and the regulation of cAMP in hepatomas, we have examined the cell line MH1C1 from Morris hepatoma 7795. These cells in culture contained low intracellular cAMP concentrations (approximately 0.5 pmol/mg protein at confluency), and were unresponsive to glucagon and prostaglandins (PG) E1 and E2. In contrast, solid hepatomas in rats developed from inoculates of MH1C1 had a 40-fold higher basal cAMP concentration and were stimulated by PGE1 and PGE2. Fibroblasts cultured from these tumours also contained high cAMP levels and responded strongly to PGE1. This may suggest that the difference in cAMP regulation between hepatomas in vivo and hepatoma cells in vitro results from the presence of other cells in the solid tumour rather than from selection of low-cAMP cells during the cloning procedure. Low-Km and intermediate-Km cAMP phosphodiesterase activity was high in MH1C1, compared to normal hepatocytes. This might contribute to the low cAMP level. The ability of MH1C1 to form cAMP was not defective, as the level could be increased more than 200-fold by beta-adrenergic activation in the presence of the phosphodiesterase inhibitor methylisobutylxanthine. PMID- 3035997 TI - Disorders of the autonomic nervous system: Part 1. Pathophysiology and clinical features. AB - Autonomic dysfunction may result from diseases that affect primarily either the central nervous system or the peripheral autonomic nervous system. The most common pathogenesis of disturbed autonomic function in central nervous system diseases is degeneration of the intermediolateral cell columns (progressive autonomic failure) or disease or damage to descending pathways that synapse on the intermediolateral column cells (spinal cord lesions, cerebrovascular disease, brainstem tumors, multiple sclerosis). The peripheral autonomic nervous system may be damaged in isolation in the acute and subacute autonomic neuropathies or in association with a generalized peripheral neuropathy. The peripheral neuropathies most likely to cause severe autonomic disturbance are those in which small myelinated and unmyelinated fibers are damaged in the baroreflex afferents, the vagal efferents to the heart, and the sympathetic efferent pathways to the mesenteric vascular bed. Acute demyelination of the sympathetic and parasympathetic nerves in the Guillain-Barre syndrome may also cause acute autonomic dysfunction. Although autonomic disturbances may occur in other types of peripheral neuropathy, they are rarely clinically important. PMID- 3035998 TI - Inactivation of GM1-ganglioside beta-galactosidase by a specific inhibitor: a model for ganglioside storage disease. AB - This study was designed to establish an in vitro model with biochemical and morphological similarities to the human neurodegenerative disease GM1 gangliosidosis. Utilizing a specific inactivator of the lysosomal enzyme GM1 ganglioside beta-galactosidase (beta-D-galactopyranosylmethyl-p nitrophenyltriazene [beta-GalMNT]) and neuroblastoma X glioma hybrid cells (NG108 15), we suppressed beta-galactosidase activity for up to 72 hours. Coincidental with suppression of this enzyme to levels less than 1% of control, we found up to a nine-fold accumulation of its substrate, the GM1-ganglioside, and the ultrastructural appearance of membranous cytoplasmic bodies. beta-GalMNT treatment suppressed growth but had little effect on the specific activity of choline acetyltransferase, lactate dehydrogenase, or other lysosomal enzymes including galactosylceramidase. This model should permit studies of the neurophysiological effects of increased ganglioside accumulation and their reversibility. PMID- 3035999 TI - The role of taste in rapid sodium satiation by sodium-deficient sheep. AB - Sheep with a parotid fistula and sodium-deprived for 24 or 48 h (Na deficit = 500 700 mmol) were trained to drink their entire requirement of sodium bicarbonate solution from a cup in their cage in a single draught for up to 2 min. The cup was connected to a reservoir by an apparatus that enabled the concentration of the solution offered to be changed after the animal had drunk the first 100 or 150 ml of fluid without interrupting the flow of fluid or disturbing the drinking sheep. Under control conditions, the concentrations of solutions in the cup and reservoir were the same, either 900 mM or 300 mM NaHCO3. On experimental days, the concentration of NaHCO3 in the cup and reservoir were different so that the concentration of fluid increased from 300 mM to 900 mM or decreased from 900 mM to 300 mM NaHCO3. On those experimental days when the concentration of NaHCO3 was increased from 300 to 900 mM, the sheep drank a volume of fluid sufficient to maintain intake commensurate with loss. However, when the concentration of NaHCO3 was decreased from 900 to 300 mM, the sheep drank a volume of fluid insufficient to correct the deficit. It is proposed that the failure of sheep to react appropriately to a decrease in NaHCO3 concentration is a consequence of taste adaptation. PMID- 3036000 TI - Mixed enzymic reaction--internal diffusion kinetics of nonuniformly distributed immobilized enzymes. The system agarose-micrococcal endonuclease. AB - Two types of (CNBr-activated) Agarose-staphylococcal endonuclease derivatives have been prepared, one with the enzyme uniformly distributed in the support, and the other with the enzyme preferentially bound in the most external part of the support particles; the latter were obtained using agarose of very small pores and a high degree of activation. Quantitative enzyme distribution has been determined by scanning fluorescence microscopy. With these insoluble enzyme derivatives, a kinetic study for the hydrolysis of a mononucleotide has been carried out. A simple theoretical model for nonuniformly distributed insoluble enzyme derivatives, which considers only the case of mixed enzymic reaction-internal diffusion kinetics, is proposed. The experimental data agree very well with the predictions of the model. PMID- 3036001 TI - The action of sphingomyelinase from Bacillus cereus on ATP-depleted bovine erythrocyte membranes and different lipid composition of liposomes. AB - The presence of cholesterol or phosphatidylethanolamine in sphingomyelin liposomes enhanced 2- to 10-fold the breakdown of sphingomyelin by sphingomyelinase from Bacillus cereus. On the other hand, the presence of phosphatidylcholine was either without effect or slightly stimulative at a higher molar ratio of phosphatidylcholine to sphingomyelin (3/1). In the bovine erythrocytes and their ghosts, the increase by 40-50% or the decrease by 10-23% in membranous cholesterol brought about acceleration or deceleration of enzymatic degradation of sphingomyelin by 50 or 40-50%, respectively. The depletion of ATP (less than 0.9 mg ATP/100 ml packed erythrocytes) enhanced K+ leakage from, and hot hemolysis (lysis without cold shock) of, bovine erythrocytes but decelerated the breakdown of sphingomyelin and hot-cold hemolysis (lysis induced by ice-cold shock to sphingomyelinase-treated erythrocytes), either in the presence of 1 mM MgCl2 alone or in the presence of 1 mM MgCl2 and 1 mM CaCl2. Also, ATP depletion enhanced the adsorption of sphingomyelinase onto bovine erythrocyte membranes in the presence of 1 mM CaCl2 up to 81% of total activity, without appreciable K+ leakage and hot or hot-cold hemolysis. These results suggest that the presence of cholesterol or phosphatidylethanolamine in biomembranes makes the membranes more susceptible to the attack of sphingomyelinase from B. cereus and that the segregation of lipids and proteins in the erythrocyte membranes by ATP depletion causes the deceleration of sphingomyelin hydrolysis despite the enhanced enzyme adsorption onto the erythrocyte membranes. PMID- 3036002 TI - Production of 4-hydroxypyrazole from the interaction of the alcohol dehydrogenase inhibitor pyrazole with hydroxyl radical. AB - Pyrazole, an effective inhibitor of alcohol dehydrogenase, was previously shown to be a scavenger of the hydroxyl radical. 4-Hydroxypyrazole is a major metabolite in the urine of animals administered pyrazole in vivo. Experiments were conducted to show that 4-hydroxypyrazole was a product of the interaction of pyrazole with hydroxyl radical generated from three different systems. The systems utilized were the iron-catalyzed oxidation of ascorbate, the coupled oxidation of hypoxanthine by xanthine oxidase, and NADPH-dependent microsomal electron transfer. Ferric-EDTA was added to all the systems to catalyze the production of hydroxyl radicals. A HPLC procedure employing either uv detection or electrochemical detection was utilized to assay for the production of 4 hydroxypyrazole. The three systems all supported the oxidation of pyrazole to 4 hydroxypyrazole by a reaction which was sensitive to inhibition by competitive hydroxyl radical scavengers such as ethanol, mannitol, or dimethyl sulfoxide and to catalase. The sensitivity to catalase implicates H2O2 as the precursor of the hydroxyl radical by all three systems. Superoxide dismutase inhibited production of 4-hydroxypyrazole only in the xanthine oxidase reaction system. In the absence of ferric-EDTA (and azide), microsomes catalyzed the oxidation of pyrazole to 4 hydroxypyrazole by a cytochrome P-450-dependent reaction which was independent of hydroxyl radicals. This latter pathway may be primarily responsible for the in vivo metabolism of pyrazole to 4-hydroxypyrazole. The production of 4 hydroxypyrazole from the interaction of pyrazole with hydroxyl radicals may be a sensitive, rapid technique for the detection of these radicals in certain tissues or under certain conditions, e.g., increasing oxidative stress. PMID- 3036003 TI - The oxidation of NADH by tetravalent vanadium. AB - Vanadyl (V(IV)) salts autoxidize in neutral aqueous solution yielding O2- plus vanadate (V(V)) and these, in turn, cause the oxidation of NADH, by a free radical chain reaction. This oxidation of NADH was inhibited by superoxide dismutase, but not by a scavenger of HO.. When H2O2 was present V(IV) caused rapid oxidation of NADH by a process which was unaffected by superoxide dismutase but was inhibited by a scavenger of HO.. This appeared to be dependent upon reduction of H2O2 to OH- plus HO., by V(IV)), followed by oxidation of NADH by HO.. Since there are reductants, within cells, capable of reducing V(V) to V(IV), these reactions are likely to contribute to the toxicity of vanadate. PMID- 3036004 TI - Negative and positive assays of superoxide dismutase based on hematoxylin autoxidation. AB - Hematoxylin, a natural dye commonly used as a histological stain, generates superoxide upon oxidation to its quinonoid product, hematein. The parameters affecting this reaction were assessed in developing a new and versatile assay for superoxide dismutase. The autoxidation of hematoxylin to hematein was accompanied by an increase in absorbance between 400 and 670 nm. The autoxidation rate was proportional to hematoxylin concentration and increased with pH above 6.55. Trace metals accelerated the autoxidation and this effect was eliminated by EDTA. Superoxide dismutase inhibited the autoxidation 90-95% below pH 7.8, but above pH 8.1 the rate was augmented by superoxide dismutase. The rate inhibition at low pH was proportional to the superoxide dismutase concentration up to 70% inhibition. The rate acceleration at high pH was proportional to superoxide dismutase concentration up to approximately 200% acceleration. The autoxidation rate was not significantly affected by ethanol, cyanide, azide, hydrogen peroxide, or catalase. However, the reaction was inhibited by the reducing agents NADH, reduced glutathione, ascorbate, and dithiothreitol, and by undialyzed extracts of Escherichia coli B. When cell extracts were dialyzed prior to assay, the degree of inhibition observed was proportional to the concentration of superoxide dismutase in the extract. These observations form the basis for negative and positive assays of superoxide dismutase which are inexpensive and simple to perform. The negative assay has the added advantage of being applicable at physiological pH. PMID- 3036006 TI - Generation of radical anions of nitrofurantoin, misonidazole, and metronidazole by ascorbate. AB - Nitrofurantoin, misonidazole, and metronidazole were reduced to their corresponding nitro anion radicals by ascorbate in anaerobic solutions at high pH. The nitrofurantoin anion radical could be detected at neutral pH. In neutral solutions, the nitro anion radicals of misonidazole and metronidazole were too unstable to be observed by electron spin resonance spectroscopy. At neutral pH, solutions containing ascorbate, nitrofurantoin, or misonidazole consumed oxygen. The addition of superoxide dismutase, catalase, or both superoxide dismutase and catalase decreased the rate of oxygen consumption. These results show that nitro anion radicals are formed by reduction with ascorbate, and superoxide anion radical and hydrogen peroxide are produced by reactions of these radicals with oxygen. PMID- 3036005 TI - The cyclic nucleotide-dependent phosphorylation of aortic smooth muscle membrane proteins. AB - Membrane proteins of Mr 240,000, 130,000, and 85,000 (GS-proteins) were rapidly and selectively phosphorylated in particulate fractions of rabbit aortic smooth muscle in the presence of [Mg-32P]ATP and low concentrations of cGMP (Ka = 0.01 microM) or cAMP (Ka = 0.2 microM). The effects of both cyclic nucleotides in this preparation were mediated entirely by an endogenous, membrane-bound form of cGMP dependent protein kinase (G-kinase). The GS-proteins were also phosphorylated by the soluble form of G-kinase purified from bovine lung; this effect was most evident following removal of endogenous G-kinase from the membranes using Na2CO3 and high salt washes. The membrane-bound and cytosolic forms of G-kinase phosphorylated the Mr 130,000 GS-protein with the same specificity as determined by two-dimensional peptide mapping. Despite this functional homology between the two forms of G-kinase, only the particulate enzyme appears to play a role in phosphorylating the GS-proteins. Although little endogenous cAMP-dependent protein kinase (A-kinase) activity was detected in washed aortic smooth muscle membranes, the GS-proteins could be phosphorylated when purified A-kinase catalytic subunit was added to this preparation. Peptide mapping of the Mr 130,000 GS-protein indicated that A-kinase phosphorylated a subset of the same peptides labeled by the two forms of G-kinase. The endogenous A-kinase of rabbit aortic smooth muscle homogenates was also found to phosphorylate the GS-proteins. Since the intracellular concentrations of cGMP or cAMP can be selectively elevated by different stimuli, these results suggest several possible mechanisms by which the phosphorylation state of the GS-proteins may be regulated by cyclic nucleotides: activation of the membrane-bound G-kinase by cGMP or cAMP; and activation of cytosolic A-kinase by cAMP. PMID- 3036007 TI - Evidence for the involvement of an acidic compartment in the processing of myeloperoxidase in human promyelocytic leukemia HL-60 cells. AB - The observation that myeloperoxidase precursor and larger intermediate (Mr 91,000 and 81,000, respectively) were extracted in the presence of detergent from isolated granule fractions of human promyelocytic leukemia HL-60 cells under mildly acidic conditions was investigated. In contrast, under conditions of neutral pH, only the Mr 74,000 intermediate and mature species were extracted. Extraction of the Mr 91,000 and 81,000 forms was also enhanced in the presence of EDTA. Kinetic studies of the processing of the different myeloperoxidase species confirmed the intermediate nature of the Mr 81,000 and 74,000 forms. Support for a role of an acidic intracellular compartment was obtained through evidence that the acid-extractable precursor and intermediates accumulated in HL-60 cells which had been treated with 1 microM monensin. Under these conditions, the production of mature heavy (Mr 63,000) and light (Mr 13,500) subunits of myeloperoxidase was consistently inhibited by greater than 40% over a 16-h period. The effects of monensin on processing of myeloperoxidase were completely reversed if monensin was removed during this 16-h period. These data support the idea that an acidic compartment may be involved in the transport of myeloperoxidase precursors to azurophil granules and/or their processing to a smaller intermediate form (Mr 74,000) of the enzyme. PMID- 3036008 TI - Single-strand-specific nuclease of pea seeds: glycoprotein nature and associated nucleotidase activity. AB - A single-strand-specific nuclease from germinating pea seeds has been purified to homogeneity. The purification procedure includes affinity chromatography on concanavalin A-Sepharose and gel filtration. The nuclease exhibits its activity at neutral pH and does not have an absolute requirement for a divalent cation. The purified nuclease also possesses a 3'-nucleotidase activity and is a glycoprotein containing about 20% carbohydrate. On native polyacrylamide gels the nuclease activity comigrates with the nucleotidase. Sodium dodecyl sulfate polyacrylamide gel electrophoresis showed the presence of two subunits in the native enzyme. The nuclease and nucleotidase activities show differential rates of thermal inactivation, the latter following simple first order kinetics and the former exhibiting a more complex reaction. The nucleotidase was also found to be stimulated by DNA, the increase being greater with native DNA than with denatured DNA. These properties are possibly accounted for by the dimeric structure of the enzyme where the nucleotidase catalytic site resides in one subunit while the nuclease site is formed by interaction of both subunits. The enzyme also hydrolyzes double-stranded alkylated DNA and depurinated DNA at a higher rate than native DNA. Experimental evidence suggests that depurinated DNA is hydrolyzed in the region of apurinic sites. PMID- 3036009 TI - Calmodulin participation in oxygen radical-induced cardiac sarcoplasmic reticulum calcium uptake reduction. AB - The effect of scavengers of oxygen radicals on canine cardiac sarcoplasmic reticulum (SR) Ca2+ uptake velocity was investigated at pH 6.4, the intracellular pH of the ischemic myocardium. With the generation of oxygen radicals from a xanthine-xanthine oxidase reaction, there was a significant depression of SR Ca2+ uptake velocity. Xanthine alone or xanthine plus denatured xanthine oxidase had no effect on this system. Superoxide dismutase (SOD), a scavenger of .O2-, or denatured SOD had no effect on the depression of Ca2+ uptake velocity induced by the xanthine-xanthine oxidase reaction. However, catalase, which can impair hydroxyl radical (.OH) formation by destroying the precursor H2O2, significantly inhibited the effect of the xanthine-xanthine oxidase reaction. This effect of catalase was enhanced by SOD, but not by denatured SOD. Dimethyl sulfoxide (Me2SO), a known .OH scavenger, completely inhibited the effect of the xanthine xanthine oxidase reaction. The observed effect of oxygen radicals and radical scavengers was not seen in the calmodulin-depleted SR vesicles. Addition of exogenous calmodulin, however, reproduced the effect of oxygen radicals and the scavengers. The effect of oxygen radicals was enhanced by the calmodulin antagonists (compounds 48/80 and W-7) at concentrations which showed no effect alone on Ca2+ uptake velocity. Taken together, these findings strongly suggest that .OH, but not .O2-, is involved in a mechanism that may cause SR dysfunction, and that the effect of oxygen radicals is calmodulin dependent. PMID- 3036011 TI - [Preoperative radiotherapy of brain tumors]. AB - Irradiation prior to radical surgical removal has been performed 51 times in 48 cases of brain tumor over the last 7 years. Through this procedure, all cases have attained good results in both clinical and CT evaluations. In the course of radiotherapy, various doses of corticosteroids and/or ventricular drainage were applied to prevent dangerous brain swelling and hydrocephalus. Total radiation doses were 40-50 Gy in primary cases and 30-40 Gy in recurrent cases. Through this preoperative radiotherapy, tumors showed reduction of their feeding vessels (78.9%) on angiography and/or became necrotized with cyst formation. This facilitated case of surgery and permitted radical resection of these tumors (41%). Especially, in cases of germ cell tumors of mixed type and medulloblastomas, this procedure permitted radical resection in 75% and 100% of cases, respectively. Among glioblastomas and grade 3 astrocytomas, 21.7% were radically resected and the patients involved showed a good relative survival rate. This method may actually facilitate radical cure of malignant brain tumors soon. PMID- 3036012 TI - [Treatment of malignant fibrous histiocytoma of soft tissues]. AB - Thirty-six patients with soft tissue malignant fibrous histiocytoma were followed up. At the time of this study, 14 of the 36 patients were alive and the average follow-up period was 59.3 months. The average age at initial diagnosis was 62.1 years. Of the 36 patients, 13 were initially treated by us. All of the marginal excision cases (17) showed recurrence. Five of 8 additional excision cases had residual tumor and three of these showed recurrence. Those cases given primary wide excision (5) showed no local recurrence. One out of 3 amputation cases showed recurrence five years after the operation. Thus, the total recurrence rate was 64%. The average period of recurrence was 9.3 months (n = 21). Radiotherapy was effective in four patients among 11 irradiated cases. Chemotherapy is difficult to apply, since patients are generally too old to tolerate intensive chemotherapy and also the drug-sensitivity is low. For this reason, we have insufficient clinical experience to make an evaluation. Five-year survival rate was 45% and 10-year survival, 30%. Early detection of the tumor and adequate initial surgery are the best ways of improving the results of treatment for MFH. Chemotherapy and radiotherapy might achieve adjuvant effects and produce better results. PMID- 3036013 TI - [Chemoembolization]. AB - Chemoembolization is a technique by which the blood flow in the artery feeding a tumor is arrested and, at the same time, an antitumor agent is delivered in a high concentration to the target site in anticipation of a synergistic antitumor effect. Usually, this is a transcatheter technique. The embolic materials used to arrest the blood flow include gelatin sponge, Lipiodol, microcapsule, albumin microsphere, degradable starch microsphere and the like. Since the gelatin sponge and Lipiodol are available on the market, transcatheter oily chemoembolization (TOCE) using these two materials was performed in cases of hepatic tumor. In many cases of TOCE, adriamycin was used as an adriamycin solution Lipiodol mixture (adriamycin-in-oil emulsion). The cumulative survival rates for 100 patients with unresectable hepatoma treated by TOCE were 53.8% for one year and 36.5% for two years. Thus, improvement was observed in comparison with the cumulative survival rates of 104 patients who underwent hepatic embolization without Lipiodol (1 year, 45.2%, 2 years, 16.3%). Adriamycin-in-oil emulsion retained in the tumor as microemboli brings about the slow-releasing effect of adriamycin. The effect was demonstrated in the blood and tissue concentrations of adriamycin following TOCE. PMID- 3036010 TI - [Qualitative improvement of the surgical treatment of cancer using laser equipment--surgical technic after photodynamic therapy]. AB - Nineteen patients with lung cancer were treated by combined preoperative photodynamic therapy (PDT) and surgery. Preoperative photodynamic therapy was performed for the purpose of either reducing the extent of resection or increasing operability. Clinically, nine patients had stage I disease, one had stage II, eight had stage III and one had stage IV. There were two cases of tracheal superficial invasion from primary lesions, three cases of intrabronchial polypoid tumor or superficial invasion of the carina by primary lesions, eleven cases of polypoid tumor or superficial invasion of the main bronchus, and three cases of double primary lesions. Argon dye laser was used in this study. Preoperative PDT was performed 48 to 72 hours after intravenous injection of hematoporphyrin derivative (HpD). Therapeutic conditions were 60 to 600 joules/cm2 for the superficial invasive areas and an additional 200 to 800 m W for 8 to 15 minutes for polypoid lesions. Surgical resection was performed 1 to 9 weeks after PDT. The initial purpose of PDT was achieved in 15 of the 19 patients treated. In five of six originally inoperable cases, conversion to an operable status was achieved. Thirteen patients were originally candidates for pneumonectomy, and it became possible to reduce the extent of resection to lobectomy in ten of them. This study suggests that PDT may have an important role in combination with surgery and other modalities in advanced lung cancers. PMID- 3036014 TI - [Recent status of the diagnosis and treatment of bone metastasis in patients with advanced lung cancer]. AB - The incidence and prognosis of patients with bone metastasis in primary advanced lung cancer were studied retrospectively. Between Jan. 1980 and Dec. 1985, 289 cases entered various kinds of chemotherapy protocol studies. Patients with bone metastasis of non-small cell lung cancer (NSC) comprised 44% (86/192), and those with small cell lung cancer (SC) comprised 43% (42/97). Histologically, 48% of adenocarcinoma, 50% of large cell carcinoma and 31% of squamous cell carcinoma showed bone metastasis. 8 percent of NSC bone meta (+) cases had an initial symptom of bone metastasis. Bone scan and bone X-ray were complementary and useful for diagnosis of bone metastasis, and sequential examinations tended to reduce the incidence of false-positive cases. Vertebral column, rib, pelvis and femur were the most common sites. Over 70% of the bone metastasis were in multiple skeletal systems, and 90% showed multiple-site involvement for both NSC and SC. Radiation therapy effectively reduced severe pain but paralysis was hard to control. In very few cases surgical treatment was indicated because of multiple bone metastasis, and systemic dissemination. Bone scan in 12% of SC patients showed apparent improvement with systemic chemotherapy. Among the M1 group of adenocarcinoma, median survival was 9 months in bone (+) cases, 11 months in bone (-) cases, 2 year survival was 8%, and 24%, and 3-year survival 2% and 22%, respectively. Among the bone(+) group and bone(-) group in ED cases of SC, median survival was 10 months vs. 11 months, and 2-year survival rates were both 13%. 22 percent (8/36) of squamous cell carcinomas without bone metastasis showed hypercalcemia (5.5 mEq/l). In patients with advanced lung cancer the major goal of treatment is recovery of the performance status of the patient and the relief of pain. In the case of SC, intensive systemic chemotherapy should be conducted as an adjuvant to local therapy. PMID- 3036015 TI - [Bone metastasis of malignant solid tumors in childhood]. AB - A total of 452 cases of childhood malignant solid tumors were treated over the last twenty years at the National Children's Hospital. These included 175 cases of neuroblastoma, 64 cases of Wilms' tumor, 65 cases of malignant lymphoma, 45 cases of soft tissue sarcoma, 31 cases of hepatoma, 20 cases of malignant teratoma, 17 cases of testicular tumor, 7 cases of ovarian tumor and 28 cases of other forms of malignant solid tumor. Bone metastasis was observed in 62 of 175 cases of neuroblastoma, 3 of 64 cases of Wilms' tumor, one of 65 cases of malignant lymphoma, 4 of 45 cases of soft tissue sarcoma, one case of pulmonary blastoma and one case of osteogenic sarcoma, giving a total occurrence of bone metastasis in 72 of the 452 cases. The main sites of bone metastasis in neuroblastoma were the skull (61.4%), femur (56.8%), orbit (27.3%) and spine (22.7%). The average values of serum calcium and alkaline phosphatase activity showed no significant difference. The patients with bone metastasis were treated with a combination of radiation therapy and intensive chemotherapy, resulting in temporary improvement. The survival of patients with stage IV neuroblastoma with bone metastasis was worse than that of similar patients without bone metastasis. PMID- 3036016 TI - [UFT in the treatment of primary lung cancer--5-FU concentration in the tissue and side effects]. AB - The concentrations of FT, 5-FU and uracil in lung cancer tissue, healthy lung tissue and serum were measured in 10 patients with primary lung cancer to whom UFT (600 mg) had been administered for one week. No remarkable difference was found in the concentration of FT between both tissues. The concentration of 5-FU in lung cancer tissue was 0.099 +/- 0.051 microgram/g and significantly higher than the level of 0.009 +/- 0.003 microgram/ml in serum (p less than 0.001) and 0.039 +/- 0.021 microgram/g in normal lung tissue (p less than 0.01). The ratio of 5-FU concentration in cancerous tissue to that in the serum was 11.0, while the ratio of the level in cancerous tissue to that in normal lung tissue was 2.5. The concentration of uracil was much higher in cancerous tissue than in normal lung tissue and serum. Maintenance therapy with UFT proved possible for a long period in 11 primary lung cancer patients with negligible digestive side effects. PMID- 3036017 TI - Cytomegalovirus infection in day nurseries. AB - One hundred and seventeen children and 41 teachers in day nurseries were screened for cytomegalovirus (CMV) viruria over a period of one year. Thirty two (27%) children and two (5%) teachers were found to be excreting virus on at least one occasion. Restriction endonuclease typing showed that virus strains isolated from the children were dissimilar, with the exception of those from sibling pairs and one unrelated pair. The virus isolate from one teacher matched those from two unrelated children, while the isolate from another teacher could not be distinguished from that from a sibling pair. The CMV serological state of the 41 teachers was not significantly different from 500 matched controls and no seroconversions occurred. It is concluded that although transmission of CMV among children and teachers may occur in day nurseries, the dissimilarity of most of the virus strains indicates that infection predominantly occurs outside. Furthermore, teachers in day nurseries showed no evidence of an increased risk of past CMV infection when compared with matched controls. PMID- 3036018 TI - Influence of antacid and formulation on effectiveness of pancreatic enzyme supplementation in cystic fibrosis. AB - A series of treatment trials, involving food balances based on determination of fat coefficient absorption, nitrogen faecal loss, and daily faecal weight, was performed in 82 patients with cystic fibrosis. Results showed that a conventional powdered pancreatic extract (Pancrex V) required a high dosage to achieve reasonable improvement in fat and nitrogen absorption (200 mg/kg body weight/day on average) and rarely restored digestion to normal. Bicarbonate (5.2 g/m2 body surface/day) slightly enhanced the enzymatic activity of the powdered extract, this being more apparent in those with more severe steatorrhoea. There was no advantage in providing the extract in microgranules protected by cellulose acetatephthalate. A product based on fungal lipase and protease (Krebsilasi) proved to be ineffective in correcting fat and protein absorption. The two recent products prepared in pH sensitive microspheres (Pancrex V microspheres and Pancreas-Prolipase) had similar advantages in digestive activity. Compared with the traditional preparations, they offered a number of practical advantages, including a smaller number of capsules (particularly Pancrex V microspheres) and improved palatability. PMID- 3036019 TI - Local production of rotavirus specific IgA in breast tissue and transfer to neonates. AB - Rotavirus specific IgA, secretory component, and IgG were measured by enzyme linked immunosorbent assay in 20 pairs of mothers and babies to estimate antibody transfer from the mother, particularly from breast milk to neonatal faeces. Colostrum contained high titres of specific IgA and secretory component, which decreased gradually. Faeces after breast feeding for three days showed detectable titres of IgA and secretory component, with further increases by seven days. There was a positive correlation between titres of secretory component in breast milk and in faeces. To clarify the mechanism of high anti-rotavirus activity in breast milk, ratios of rotavirus specific IgA in maternal serum samples to breast milk were calculated and compared with those that were herpes simplex virus specific. Significantly higher concentrations were obtained for the herpes simplex virus specific samples, indicating that anti-rotavirus IgA is selectively produced in breast tissue. PMID- 3036020 TI - Nedocromil sodium and exercise induced asthma. AB - Serial exercise tests were carried out by 12 children with asthma on two study days. After a control exercise test either nedocromil sodium 4 mg or placebo were given double blind by metered dose inhaler. Highly significant inhibition of exercise induced asthma occurred after nedocromil, lasting for over two hours. PMID- 3036021 TI - Serological evidence of herpes simplex virus infection in atopic eczema. AB - Twenty three of 113 patients (20%) with atopic eczema had neutralising antibodies to herpes simplex virus compared with 34 of 113 matched controls (30%), an insignificant difference. This suggests that children with atopic eczema are no more likely to acquire herpes simplex infection than normal children. PMID- 3036022 TI - Group B streptococcal infection presenting as sudden death in infancy. AB - Two infants aged 3 and 8 months were found dead in their cots. Postmortem cultures yielded group B streptococci from multiple internal sites, and cytomegalovirus was also isolated from the lungs of one. These cases show the value of microbiological culture in the investigation of sudden infant death. PMID- 3036023 TI - Topical treatment of psoriasis with the topoisomerase inhibitors novobiocin and nalidixic acid: a pilot study. AB - Our studies in human epidermal keratinocytes as a model system have suggested that the antibiotic topoisomerase II inhibitors, novobiocin and nalidixic acid, may be of value for the treatment of hyperproliferative skin disorders. We have therefore conducted a pilot study of the clinical efficacy of these compounds for the treatment of psoriasis. The compounds were administered topically to psoriatic plaques in seven healthy patients over a period of 6 weeks. Nalidixic acid (2%) or novobiocin (2% or 5%) in methylcellulose were applied twice daily under occlusion, and methylcellulose alone was used as a control. In six of the seven patients, one or both compounds effected somewhat greater improvement than in the control within 3 weeks of treatment. PMID- 3036024 TI - Collagen degradation in the pregnant human cervix at term and after prostaglandin induced cervical ripening. AB - The role of enzymatic collagen degradation in prostaglandin-induced and physiological cervical ripening was studied by determining collagenase and other proteolytic activity in extracts of cervical biopsies. Collagenase activity was assayed in a highly specific and sensitive system using native collagen type I as substrate. The intracervical application of sulprostone gel prior to termination of 1st trimester pregnancy led to a marked improvement in cervical dilatability. Collagenase and proteolytic activity were found to be significantly higher in postpartum samples than in specimens obtained from the nonpregnant or early pregnant cervix. After sulprostone treatment enzymatic activities were only marginally elevated. Analysis of extractable peptides showed that sub partu collagen was degraded in preference to noncollagenous proteins into very small fragments, whereas in the process of prostaglandin-induced cervical ripening collagen degradation appears to be of minor importance. PMID- 3036026 TI - Retinoid modulation of collagenase production by adherent human mononuclear cells in culture. AB - Previous observations have suggested that retinoids might be useful for the treatment of rheumatoid arthritis. In this study we examined the effects of various retinoids on collagenase production by adherent human peripheral blood mononuclear cells in culture. We have previously shown that these cells, consisting predominantly of monocyte-macrophages, actively synthesize and secrete collagenase upon stimulation with concanavalin A. The cells were incubated in serum free medium with all-trans-retinoic acid, 13-cis-retinoic acid, all-trans retinal, or Ro 10-9359 (trimethylmethoxyphenyl retinoic acid ethyl ester) for up to 72 hours, and the collagenase activity was determined with [3H]proline labelled type I collagen as substrate. The incubation of mononuclear cells with all-trans-retinoic acid in the concentration range 10(-7)-10(-5) mol/l resulted in a dose dependent inhibition of the collagenase production. All-trans-retinal was also a potent inhibitor, whereas 13-cis-retinoic acid and Ro 10-9359 in a concentration of 10(-5) mol/l had a lesser effect. Control experiments indicated that the inhibition of collagenase production by all-trans-retinoic acid did not result from inhibition of total protein synthesis nor could it be explained by induction of an inhibitory molecule. These results indicate that retinoids with distinct structural features can inhibit collagenase production by monocyte macrophages, and suggest a role for retinoids in the treatment of rheumatoid arthritis. PMID- 3036025 TI - Molecular basis of activation and regulation of the phagocyte respiratory burst. AB - The molecular basis of activation and regulation of the phagocyte respiratory burst is discussed with particular reference to the role of inositol phospholipid hydrolysis, guanine nucleotide coupling proteins, and activation of protein kinase C. PMID- 3036027 TI - Urgent treatment for nonresectable, asphyxiating tracheal cylindroma. AB - A 22-year-old woman with an irresectable tracheal cylindroma asphyxiated during radiotherapy. Extracorporeal oxygenation was used to facilitate endotracheal resection and the insertion of a tracheobronchial stent. PMID- 3036029 TI - Antihypertensive activity of a new ACE-inhibitor, SCH 33844, during repeated administration in spontaneously hypertensive rats. AB - The antihypertensive activity of SCH 33844, a new angiotensin converting enzyme inhibitor, was investigated in conscious spontaneously hypertensive rats during a 3-week treatment regimen and for 1 week following drug withdrawal. SCH 33844 was given once daily at 2 dose levels (1 and 5 mg/kg orally) and its effects were compared with those of captopril (60 mg/kg orally). Systolic blood pressure was recorded twice weekly just before administration and at varying time intervals up to 6 hr after dosing; recordings were continued for 1 week after drug withdrawal. SCH 33844 was found to produce dose-related antihypertensive effects. Given at 1 mg/kg, the compound elicited small but significant blood pressure changes during the treatment. After drug withdrawal, systolic blood pressure returned to pre drug levels within 1 week. SCH 33844 at 5 mg/kg, and captopril at 60 mg/kg, both reduced blood pressure markedly and to a similar extent. After each administration the effect was rapid in onset, lasted for over 6 hr and was not subject to tolerance. Drug withdrawal resulted in a gradual return of systolic blood pressure toward pre-treatment levels within 1 week, with no evidence of rebound phenomena. These results indicate that SCH 33844, like captopril, produces an effective antihypertensive action throughout a repeated dosing. PMID- 3036028 TI - Angiotensin converting enzyme inhibitory activity of SCH 33844 (spirapril) in rats, dogs and monkeys. AB - SCH 33844 is a new non-sulfhydryl-containing angiotensin converting enzyme (ACE) inhibitor. SCH 33844 diacid inhibited hydrolysis of the synthetic substrate hippuryl-histidyl-leucine by rabbit lung ACE in vitro with an IC50 (concentration inhibiting enzyme by 50%) of 0.81 nM. The ester was 83 times less active. Intravenous administration of SCH 33844 and its diacid inhibited pressor responses to angiotensin I (AI) in anesthetized rats with calculated ID50's of 16 and 8 micrograms/kg, respectively. Oral administration of SCH 33844 (0.03-1 mg/kg) inhibited AI pressor responses in conscious rats with a duration of 24 hr at the highest dose. The diacid was inactive. Intravenous administration of SCH 33844 (100-1000 micrograms/kg) or its diacid (30 micrograms/kg) to anesthetized dogs inhibited AI pressor activity and potentiated the depressor response to bradykinin. SCH 33844 inhibited AI responses in conscious dogs following oral administration of 0.3-3 mg/kg. Oral administration of SCH 33844 (1 mg/kg) to conscious monkeys inhibited AI pressor responses for the 4 hr duration of study. In conclusion, SCH 33844 is a potent, orally effective ACE inhibitor in rats, dogs and monkeys. PMID- 3036031 TI - Utility of technetium Tc 99m pyrophosphate bone scanning in cardiac amyloidosis. AB - Thirty-four patients with amyloidosis proved by biopsy specimen were studied using technetium Tc 99m pyrophosphate scintigraphy to assess its utility in the diagnosis of amyloid heart involvement. Of 14 patients studied retrospectively, only three had intense uptake judged to be diagnostic of cardiac amyloidosis. In a prospective analysis of 20 patients with amyloidosis, all of whom had evidence of cardiac involvement by two-dimensional echocardiography, 17 had abnormal scans. Fourteen of the 17 scans had only 1+ or 2+ uptake, a finding that also was present in 15 of the 20 control patients (without amyloid heart disease). Only three of the 20 patients with cardiac amyloidosis had intense uptake that was considered unequivocal and diagnostic of amyloidosis. Of the five patients with biopsy specimen proof of endomyocardial amyloidosis, only one had intense uptake and one had no uptake. When intense uptake of technetium Tc 99m pyrophosphate is found in the heart of a patient, amyloidosis is highly likely. The technique, however, is not sufficiently sensitive to warrant routine screening of patients with amyloidosis or cardiomyopathies. Cross-sectional echocardiography is superior to pyrophosphate scintigraphy for recognition of cardiac amyloidosis. PMID- 3036030 TI - Effects of forskolin on pancreatic exocrine secretion and cyclic nucleotide concentrations of the dog pancreas. AB - The effects of forskolin on the secretion of pancreatic juice and on the intracellular levels of cyclic nucleotides were investigated in preparations of the isolated and blood-perfused dog pancreas. Forskolin (3-100 micrograms) injected intra-arterially caused a dose-dependent increase in the secretion of pancreatic juice. The concentration of bicarbonate in the pancreatic juice induced by forskolin was increased. However, protein concentration was decreased. Forskolin-induced secretion was not modified by pretreatment with atropine, phentolamine, propranolol, cimetidine, sulpiride and verapamil. Forskolin (10-30 micrograms) significantly increased cyclic AMP concentration in the pancreatic tissue before the onset of the pancreatic secretion. On the other hand, cyclic GMP concentration was not influenced significantly throughout 30 min after the administration of forskolin. From these results, it is concluded that forskolin may produce an increase in pancreatic secretion mediated through an increase of intracellular cyclic AMP concentration. PMID- 3036032 TI - Lactate dehydrogenase values and bone scans as predictors of bone marrow involvement in small-cell lung cancer. AB - The value of serum lactate dehydrogenase (LDH) levels and bone scan results in predicting marrow involvement in small-cell carcinoma of the lung (SCCL) was studied. Records of 79 patients with SCCL who had undergone 92 bone marrow examinations were reviewed. None of 25 patients with marrow involvement had a normal LDH and a negative bone scan, compared with 23 of 59 in the uninvolved group. We conclude that patients with SCCL with both negative bone scans and normal LDH values have a less than 5% chance of marrow involvement. PMID- 3036033 TI - Pure red cell aplasia associated with administration of sustained-release procainamide. AB - Pure red cell aplasia developed in a patient with small-cell lung cancer who was also taking sustained-release procainamide. Shortly after discontinuation of the procainamide preparation, a reticulocytosis and increasing hemoglobin level were observed. PMID- 3036034 TI - On the role of cyclic AMP and the Fnr protein in Escherichia coli growing anaerobically. AB - The role of adenosine 3',5'-monophosphate (cAMP) and of the Fnr protein, a transcriptional regulator of anaerobic electron transport, in the expression of anaerobic respiration of Escherichia coli was investigated. Under conditions of fermentation or anaerobic respiration intracellular cAMP was formed in concentrations up to 4.6 nmol/g protein. From the enzymes of the anaerobic electron transfer chain from glycerol-3-P to fumarate only the expression of glycerol-3-P dehydrogenase (Freedberg WB, Lin ECC (1973) J Bacteriol 115:816 823), but not that of fumarate reductase required cAMP. Isolated Fnr protein, which has been suggested to be an additional site of action of cAMP under anaerobic conditions did not bind cAMP. It is concluded that cAMP in anaerobic growth like in aerobic growth acts as the effector of CRP and that catabolite repression plays an important regulatory role in anaerobic catabolism. The Fnr protein was present in constant amounts (0.06 mg/g cellular protein) and in constant molar mass (Mr 30,000) in aerobically and in anaerobically grown bacteria. This result excluded regulation of the activity of the Fnr protein by a change of concentration or by processing of the protein. PMID- 3036035 TI - Catabolite inactivation of isocitrate lyase from Saccharomyces cerevisiae. AB - A reversible carbon catabolite inactivation step is described for isocitrate lyase from Saccharomyces cerevisiae. This reversible inactivation step of isocitrate lyase is similar to that described for fructose 1,6-bisphosphatase. Addition of 2,4-dinitrophenol, nystatin or glucose to cultures, grown in ethanol as carbon source, caused a rapid loss of the isocitrate lyase and fructose 1,6 bisphosphatase activities at pH 5.5 but not at pH 7.5. These results suggest that intracellular acidification and thus a cAMP increase is involved in the catabolite inactivation mechanism of both enzymes. From results obtained by addition of glucose to yeast cultures at pH 7.5 it was concluded that others factors than cAMP can play a role in the catabolite inactivation mechanism of both enzymes. PMID- 3036036 TI - Inhibition of protein phosphorylation by chloroquine. AB - The rapid phase of fructose-1,6-bisphosphatase (FBPase) inactivation following glucose addition to starved yeast cells [reported previously] is inhibited on addition of 10 mM chloroquine (CQ) at about pH 8. This inhibition of inactivation was shown to be due to the prevention of phosphorylation of the enzyme. CQ was also found to inhibit general protein phosphorylation in the yeast cells. Glycolysis, as observed by changes in intracellular glucose-6-phosphate and extracellular glucose and ethanol concentrations, was shown to be significantly inhibited in cells treated with CQ. Similarly, a decrease in ATP concentrations was observed. However, during the early stages of phosphorylation of FBPase, levels of ATP were similar in cells containing CQ as in those without CQ. Thus, decrease in ATP levels is not thought to be significantly responsible for the inhibition of protein phosphorylation. However, the phosphorylating activity of cyclic AMP-dependent protein kinases is inhibited in vitro by relatively low concentrations of CQ. Thus, prevention of protein phosphorylation by CQ is believed to be due to inhibition of protein kinases in yeast cells. PMID- 3036037 TI - Somatosensory evoked potentials from dermatomal stimulation as an indicator of L5 and S1 radiculopathy. AB - Dermatomal somatosensory evoked potential (DSEP) results for L5 and S1 were contrasted with electromyography (EMG) results for 50 patients referred to the electrodiagnosis laboratory to document the presence of radiculopathy. Stimulation sites were over the dorsum of the foot at the distal fifth metatarsal for the S1 dermatome and at the web space of the first and second toe for the L5 dermatome. Recordings were made at PZ reference to FZ. Spinal cord or cauda responses could not be detected. Both EMG and DSEP were contrasted to myelography or lumbar computerized tomography results on 31 patients. Side-to-side amplitude differences proved too variable to be of use. Sixty-five percent of abnormal DSEP results were on the basis of side-to-side latency criterion, and 35% were on the basis of an absent unilateral response. When compared to EMG and anatomic studies DSEP showed less accuracy and sensitivity. The specificity of the two tests was similar. Using both positive EMG and anatomic studies to define radiculopathy, there were 27% false negative tests and 9% false positive tests. An 86% root level correlation was found between EMG and DSEP. Dermatomal somatosensory evoked potential studies add little to the diagnosis of radiculopathy. PMID- 3036038 TI - Resection of hepatocellular carcinoma after transcatheter arterial embolization. Reevaluation of the advantages and disadvantages of preoperative embolization. AB - Hepatocellular carcinomas (HCCs) were resected in eight patients who had preoperative transcatheter arterial embolization (TAE) and in 25 patients without preoperative TAE. Three patients in the former group had ruptured HCCs before operation. Two of the former group and three of the latter group were found to have recurrences after a follow-up of 1 1/2 years. Although preoperative TAE resulted in significantly increased tumor necrosis, it increased the risk of gangrenous change of the gallbladder, induced adhesion of the hepatoduodenal ligament, and was not effective in reducing operative blood loss or operative time if the vessel selected for TAE was inadequate. Pathologic examination revealed tumor emboli still existing in the intrahepatic veins. Daughter nodules and capsular invasion by tumor cells were not affected by TAE. Transcatheter arterial embolization seems to be effective in controlling bleeding from ruptured HCC prior to staged resection of the tumor. PMID- 3036039 TI - Localization and excision of nonpalpable breast lesions. A surgical evaluation of three methods. AB - Three methods of excising nonpalpable breast lesions have been evaluated: (1) "blind" method, using mammographic coordinates; (2) preoperative localization with the Frank needle; and (3) Frank needle localization aided by a multiperforated compression plate. Successful removal at first attempt occurred in about 80% with any method. The size of the biopsy specimens did not differ significantly among the three groups and is most probably a function of the breast volume. The failure rate was seven (2.1%) of 332 biopsies. Since three of the six repeated biopsies yielded specimens with malignancy, the persistence of a radiographically suspicious lesion on follow-up mammogram of the operated-on breast is an urgent indication for reoperation. PMID- 3036040 TI - Properties of poliovirus associated protein kinase. AB - Highly purified virulent poliovirus preparations harbour an endogenous protein kinase. Enzyme activity increases significantly upon purification of infectious virus particles from infected HeLa cells. Enzyme activity is stimulated by divalent cations. The substrate specificity and the degree of stimulation of the kinase are dependent on the nature of the divalent cations included in the assay. The preferred substrates for this kinase are the viral capsid proteins. Exogenously added proteins such as alpha-casein, phosvitin and protamine are also phosphorylated by the kinase. Moreover, these proteins enhance the phosphorylation of viral proteins. In the presence of Mg++ VP 2 and VP 0 are highly phosphorylated, while in the presence of Zn++ only VP 2 and VP 4, but not VP 0 or exogenous proteins are phosphorylated. Poliovirus associated protein kinase exhibits optimal activity at pH 7.9 in the presence of 10 mM Mg++. The Km for ATP is shown to be 40 microM. By testing different nucleotides as phosphate donors a specificity of the phosphorylation reaction for ATP is demonstrated. Phosphoamino acid analysis of hydrolysates of the substrates phosphorylated in the presence of Mg++ by thin layer electrochromatography and HPLC yielded phosphoserine and phosphothreonine from viral capsid proteins while hydrolysates of protamine yield only phosphoserine. Destabilization of the viral capsid, e.g. by preincubation at 42 degrees C for 20 minutes results in a stimulation of kinase activity. Moreover, phosphorylation of the poliovirus capsid proteins itself results in destabilization of the viral capsid. These findings suggest that phosphorylation of the viral coat proteins triggers or enhances the uncoating of poliovirus leading to the release of viral RNA. PMID- 3036041 TI - Fatty acid acylation of viral proteins in murine hepatitis virus-infected cells. Brief report. AB - The fatty acid acylation of the cell-associated virus-specific proteins of mouse hepatitis virus (A 59-strain) was studied. 3H-palmitate label was associated with E2, one of the two virion glycoproteins and its intracellular precursor gp 150. A 110 K protein, the unglycosylated apoprotein of gp 150, accumulated by tunicamycin treatment, also incorporated radiolabeled palmitic acid. The addition of fatty acid to the MHV-A 59 E 2 protein is therefore not dependent on glycosylation. PMID- 3036042 TI - Electropherotype heterogeneity within serotypes of human rotavirus strains circulating in Italy. Brief report. AB - Using solid-phase immune electron microscopy, 126 of 129 human rotavirus (HRV) strains could be serotyped directly in stools collected in Italy during the period 1981-1985. Prevalence was 70.5 per cent for serotype 1, and 13.2 per cent for each of serotypes 2 and 4. No serotype 3 strain was detected. In parallel, for 39 of 61 HRV strains tested the electropherotype of genomic RNA was successfully determined. Different electropherotypes were detected among strains of the same serotype, whereas the same electropherotype was found in HRV strains of different serotypes. Serotyping and electropherotyping of HRV strains appear to be complementary to each other, and both should be used in conjunction for epidemiological surveys. PMID- 3036043 TI - Factors influencing susceptibility of laboratory rodents to infection with mouse adenovirus strains K 87 and FL. Brief report. AB - Weanling outbred mice were more than 500 times less susceptible to orally administered mouse adenovirus (MAdV) strain K 87 than to MAdV-FL. Mouse susceptibility to MAdV-K 87 was age-dependent but not obviously host genotype dependent. Infant F 344 rats were not susceptible to infection with either MAdV strain. PMID- 3036044 TI - Viral replication in HeLa/fibroblast hybrid cells infected with human cytomegalovirus. AB - Three human hybrid cell lines were generated by the fusion of D 98OR, a HeLa cell variant, and TIG human diploid fibroblasts. Chromosome numbers of the hybrid cells fell between that of D 98OR cells and the combined chromosome number of the two cell lines, with three marker chromosomes identical to those of D 98OR cells. One hybrid, B-3, produced large amounts of human fibronectin as analyzed by immunohistochemistry. Only human cytomegalovirus (HCMV) infected B-3 cells showed positive fluorescence as detected by human antiserum to HCMV. Further cloning of B-3 by limiting dilution resulted in two cloned hybrids with markedly enhanced virus production as compared with B-3 cells. Treatment of these two clones with phorbol ester further enhanced virus production. These cloned hybrids may provide a tool to analyze host cell factors controlling the transcription and replication of HCMV. PMID- 3036046 TI - Phagocytosis of horse erythrocytes treated with equine infectious anemia virus by cultivated horse leukocytes. AB - Horse erythrocytes treated with equine infectious anemia virus hemagglutinin were phagocytized by cultivated horse leukocytes (mainly macrophage-like cells and partly polymorphonuclear cells) after incubation with fresh horse serum but not with inactivated horse serum. The phagocytosis began as soon as the erythrocytes were added to the leukocyte cultures, and the majority of the reaction proceeded within 30 minutes. Addition of antiserum showed a slightly suppressing but no enhancing effect on the phagocytosis. Phagocytosis seemed to be caused by the recognition of the third complement component on the affected RBC with the receptors on phagocytes, but not by the recognition of immunoglobulin. Since cultivated leukocytes were able to phagocytize erythrocytes which were treated with a quantity as small as 1/16 units of hemagglutinin, and since the hemagglutinin-antibody complex also could bind to erythrocytes and induced them to become phagocytized, the reaction appears to play an important role in the mechanisms of anemia and formation of sideroleukocytes in horses infected with the equine infectious anemia virus. PMID- 3036048 TI - [Germinogenic tumors of gonadal and extragonadal sites in children]. AB - Literature data are presented on histogenesis, classification, morphology, tumor markers and prognosis of germinogenic childhood tumors of gonadal and extragonadal origin. PMID- 3036047 TI - [Electron-microscopic study of neuroendocrine tumors of the lung]. AB - An electron-microscopic study was made of carcinoids, malignant carcinoids and small-cell lung cancer. These neoplasms are shown to represent one histogenetic group--neuroendocrine tumours (apudomas) of the lung. One can distinguish 3 main structural forms among them, i.e. epithelioid, sarcoma-like and symplastic. According to the ultrastructural criteria the neoplasms mentioned can be classified by their differentiation degree, as well, intermediately and poorly differentiated variants. This classification correlates well enough with the catamnesis of patients and may serve for prognosis of the disease. PMID- 3036045 TI - Complement-mediated hemolysis of horse erythrocytes treated with equine infectious anemia virus. AB - Horse erythrocytes treated with equine infectious anemia virus hemagglutinin were found to be lysed after incubation with fresh horse serum at 37 degrees C. Fresh guinea pig serum induced more efficient hemolysis than horse serum. Direct immunofluorescence test revealed the adsorption of complement factors on the surface of the erythrocytes. Calcium and magnesium ions were necessary for the hemolysis to take place. Antibody against equine infectious anemia virus enhanced the virus-induced complement-mediated hemolysis. These observations indicated that the classical pathway of complement activation was responsible for this virus-induced hemolysis and suggest the possibility that virus antigen, anti viral antibody and complement may play an important role in the genesis of the anemia of horses infected with the equine infectious anemia virus. PMID- 3036049 TI - Effect of dialyzer reprocessing methods on complement activation and hemodialyzer related symptoms. AB - The effects of different dialyzer processing methods and of reuse on complement activation and dialyzer-related symptoms were studied in 96 maintenance hemodialysis patients. New dialyzers were either unprocessed (Group 1) or machine washed with bleach and stored in formaldehyde (Group 2). Reused dialyzers were manually cleansed using the combination of bleach and formaldehyde (Group 3), or machine-washed in formaldehyde (Group 4) or peracetic acid (Group 5). Prewashed new dialyzers (Group 2) were associated with greater complement activation during dialysis when compared with unprocessed, new dialyzers (Group 1) (p less than 0.05). Reused, unbleached but formaldehyde-treated or peracetic acid-treated dialyzers (Groups 4 and 5) were associated with reduced complement activation (p less than 0.05). Complement activation was not reduced when bleach was used for reprocessing (Group 3). The percentage of patients without symptoms during dialysis was significantly greater with reused dialyzers than with new dialyzers (Groups 3 through 5 versus Groups 1 and 2; 39 versus 25%; p = 0.035). The severity of total symptoms correlated significantly (p = 0.0004) with complement activation. Our results suggest that total symptoms during dialysis are correlated with the degree of complement activation. However, trends in the data pertaining to chest pain suggest that factors other than complement activation may be important in the pathogenesis of some dialyzer-related symptoms. PMID- 3036050 TI - Dialyzer membranes: effect of surface area and chemical modification of cellulose on complement and platelet activation. AB - Using an ex vivo model, the effects of membrane composition and surface area on both the complement system (as reflected by plasma C3a levels) and platelets [as indicated by plasma concentrations of thromboxane B2 (TXB2) and platelet factor 4 (PF4)] were studied. In this model, polyacrylonitrile (PAN) was associated with less complement activation than cuprammonium cellulose (CC). A new "modified cellulose" (MC) membrane, in which a small number of the free hydroxyl groups on cellulose are substituted with a tertiary amino compound, was also associated with a low degree of complement activation, similar to that with PAN. However, the extent of hydroxyl group substitution in four MC membrane subtypes did not correlate with the reduction in complement activation. In studies using CC, the amount of generated C3a correlated with the membrane surface area, although the relationship was curvilinear. Plasma concentrations at the "dialyzer" outlet of TXB2 and PF4 were similar with CC, PAN, and MC. In studies with the MC subtypes, increasing the extent of hydroxyl group substitution paradoxically increased, albeit slightly, the amount of TXB2 generation. In studies with CC, a linear relationship between membrane surface area and TXB2 generation was found. The results suggest a dissociation between platelet and complement effects among different dialyzer membranes, and underline the importance of membrane surface area. PMID- 3036052 TI - Metastatic gestational trophoblastic disease: a study of institutional and non institutional cases at the National Hospital for Women in Philippines. PMID- 3036051 TI - Unusual manifestations of nervous system Borrelia burgdorferi infection. AB - Borrelia burgdorferi infection may show disease manifestations in different organ systems, including the skin, heart, central nervous system, peripheral nervous system, and joints. We report two hitherto unknown (to our knowledge) disease manifestations, an arteritis in the central nervous system, and a neuromyopathy of 11 years' duration. Diagnoses were made by demonstrating specific antibodies against B burgdorferi in the cerebrospinal fluid, and elevated specific antibody index. Both patients responded well to treatment, and intrathecal immunologic activity could not be demonstrated after treatment. Serologic analyses (enzyme linked immunosorbent assay) in serum and cerebrospinal fluid should be performed widely in cases with inflammatory findings in cerebrospinal fluid combined with neurologic disease of unclear origins. PMID- 3036053 TI - Mammalian cell functions mediating recombination of genetic elements: topological restraints. AB - In extending the scope of our previous analyses of the functions that mammalian cells engender to recombine elements of genetic information (Upcroft, Carter and Kidson, 1980a and b), I describe the capacity of cells to recognise differentially combinations of open-ended and closed molecules. The frequency of successful recombination, as assayed by the generation of viable SV40 genomes from appropriate substrates, was shown to decrease with the loss of free termini capable of invasion of an homologous DNA duplex and to be influenced by the presence of non-homologous flanking sequences, the length of homology and whether the sequence was internal or terminal. The effect of stabilisation with long homologous regions on recombination over short homologous regions and inter versus intra-molecular DNA sequence homologies was also investigated. PMID- 3036054 TI - A method for predicting Murray Valley encephalitis in southeast Australia using the Southern Oscillation. AB - Clinical cases of Murray Valley Encephalitis in southeast Australia have tended to occur in summer and autumn following extended periods of above average rainfall over most of eastern and northern Australia. The Southern Oscillation, an important mode of climatic fluctuations over the Indian and Pacific Oceans, is closely related to eastern and northern Australian rainfall. The Southern Oscillation has been used previously to develop methods for predicting rainfall fluctuations over Australia and their biological and economic impacts. The relationship, therefore, between the Southern Oscillation and Murray Valley Encephalitis in southeast Australia was examined. Darwin atmospheric pressure, an index of the Southern Oscillation, was found to be well below average during the autumn, winter and spring preceding the occurrence of Murray Valley Encephalitis. It is suggested that this relationship can be used to provide a simple, objective early warning system. PMID- 3036055 TI - Primary EBV infection of human umbilical cord lymphocytes and EBV genome-negative lymphoblastoid cell lines (BJAB and Ramos). AB - The appearance of Epstein-Barr virus (EBV)-associated nuclear antigen (EBNA) and induction of EBV-induced early antigen (EA) in human umbilical cord blood lymphocytes (HUCLs) and two EBV genome-negative Burkitt's lymphoma (BL) lines (BJAB and Ramos) were studied by infection with EBVs prepared from three different cell lines: marmoset cell line (B95-8) derived from infections mononucleosis, BL-derived cell line (P3HR-1) and human epithelial hybrid cell line (NPC-KT) derived from nasopharyngeal carcinoma. B95-8 virus can transform HUCLs but cannot superinfect Raji cells. P3HR-1 virus can transform HUCLs cells but cannot transform HUCLs. NPC-KT virus can transform HUCLs and can superinfect Raji cells. We have examined the time sequence of EBNA appearance and EA induction in HUCLs, BJAB cells and Ramos cells, in order to determine if three different strains of EBV differ in their abilities to infect their cells. We found that all three strains of EBV can induce EBNA in HUCLs, BJAB cells and Ramos cells. On the other hand, we found that P3HR-1 virus and NPC-KT virus can induce EA in BJAB cells and Ramos cells, but B95-8 virus cannot induce EA in their cells. PMID- 3036056 TI - Experimental infection of bulls and cows with bluetongue virus serotype 20. AB - Bluetongue virus serotype 20 (BTV20) was inoculated intradermally and subcutaneously in 4 bulls and by the intrauterine route in 8 nulliparous cows after insemination at oestrus. Viraemia was detected intermittently between 8 and 21 days after inoculation. Virus was isolated from tissue samples of 2 cows and a bull after slaughter at 14 days and from one bull at 28 days. Group reactive and type specific antibodies to BTV20 were demonstrated from 17 to 27 days after infection. No antibodies were detected in the animals slaughtered at 14 days. No clinical signs of disease were seen during the experiment and no gross or histopathological changes referable to BTV20 infection were observed post-mortem. Because of the viraemia and the production of detectable serum antibodies, gametes from these cattle would be excluded from export. PMID- 3036057 TI - Bluetongue virus serotype 20: experimental infection of pregnant heifers. AB - Three groups of 4 cows at 84 to 95 days, 100 to 160 days, and 170 to 180 days pregnant were inoculated both intradermally and subcutaneously with bluetongue virus serotype 20 (BTV20). Clinical observations and the viraemic and serological responses of the cows were followed for 9 to 17 weeks after inoculation. Viraemia developed in 9 of the 12 cows and was first detected 4 to 9 days after inoculation. Viraemia was detected for 4 to 21 days and in some animals only intermittently. The titre of the viraemia was obtained in 4 cows and ranged from detectable only, to 10(1) to 10(2.8) 50% tissue culture infecting doses per ml. Both serum neutralising and precipitating antibodies were detected in 11 of the 12 cows within 2 to 8 weeks after inoculation. No clinical responses were seen and one cow (516) did not develop a viraemia or produce detectable antibodies to the virus. The cows, calves and foetuses were necropsied following either parturition or slaughter between 200 and 270 days of pregnancy. No virus isolations were made from a wide range of tissues from the cows, calves or foetuses and no immunoglobulins or serum neutralising antibodies were detected in the serums of precolostral calves or foetuses at necropsy. No gross or histopathological lesions were seen in the cows, calves or foetuses, and there was no evidence that BTV20 crossed the bovine placenta or infected the foetus. PMID- 3036058 TI - Regulation of starch synthesis: enzymological and genetic studies. AB - Significant advances have been made with respect to elucidating the structure, the allosteric site, interactions of effectors, covalent modifications, and the amino acid sequence of the ADPG synthetase. It is hoped that in the near future, sufficient information will be obtained to enable facile manipulation of the plant tissue ADPG synthetase gene and its product. PMID- 3036060 TI - Regulation of 3-hydroxy-3-methylglutaryl-coenzyme A reductase in human hepatoma cell line Hep G2. Effects of inhibitors of cholesterol synthesis on enzyme activity. AB - Incubating Hep G2 cells for 18 h with triparanol, buthiobate and low concentrations (less than 0.5 microM) of U18666A, inhibitors of desmosterol delta 24-reductase, of lanosterol 14 alpha-demethylase and of squalene-2,3-epoxide cyclase (EC 5.4.99.7) respectively, resulted in a decrease of the HMG-CoA (3 hydroxy-3-methylglutaryl-coenzyme A) reductase activity. However, U18666A at concentrations higher than 3 microM increased the HMG-CoA reductase activity in a concentration-dependent manner. None of these inhibitors influenced directly the reductase activity in Hep G2 cell homogenates. Analysis by t.l.c. of 14C-labelled non-saponifiable lipids formed from either [14C]acetate or [14C]mevalonate during the cell incubations confirmed the sites of action of the drugs used. Beside the 14C-labelled substrates of the blocked enzymes and 14C-labelled cholesterol, another non-saponifiable lipid fraction was observed, which behaves as polar sterols on t.l.c. This was the case with triparanol and at those concentrations of U18666A that decreased the reductase activity, suggesting that polar sterols may play a role in suppressing the reductase activity. In the presence of 30 microM-U18666A (sterol formation blocked) the increase produced by simultaneously added compactin could be prevented by addition of mevalonate. This indicates the existence of a non-sterol mevalonate-derived effector in addition to a sterol dependent regulation. LDL (low-density lipoprotein), which was shown to be able to decrease the compactin-induced increase in reductase activity, could not prevent the U18666A-induced increase. On the contrary, LDL enhanced the U18666A effect, showing that the LDL regulation is not merely the result of introducing cholesterol to the cells. PMID- 3036061 TI - Effects of Ca2+ and Mg2+ on the activity of pyruvate dehydrogenase phosphate phosphatase within toluene-permeabilized mitochondria. AB - Mitochondria from rat epididymal white adipose tissue were made permeable to small molecules by toluene treatment and were used to investigate the effects of Mg2+ and Ca2+ on the re-activation of pyruvate dehydrogenase phosphate by endogenous phosphatase. Re-activation of fully phosphorylated enzyme after addition of 0.18 mM-Mg2+ showed a marked lag of 5-10 min before a maximum rate of reactivation was achieved. Increasing the Mg2+ concentration to 1.8 mM (near saturating) or the addition of 100 microM-Ca2+ resulted in loss of the lag phase, which was also greatly diminished if pyruvate dehydrogenase was not fully phosphorylated. It is concluded that, within intact mitochondria, phosphatase activity is highly sensitive to the degree of phosphorylation of pyruvate dehydrogenase and that the major effect of Ca2+ may be to overcome the inhibitory effects of sites 2 and 3 on the dephosphorylation of site 1. Apparent K0.5 values for Mg2+ and Ca2+ were determined from the increases in pyruvate dehydrogenase activity observed after 5 min. The K0.5 for Mg2+ was diminished from 0.60 mM at less than 1 nM-Ca2+ to 0.32 mM at 100 microM-Ca2+; at 0.18 mM-Mg2+, the K0.5 for Ca2+ was 0.40 microM. Ca2+ had little or no effect at saturating Mg2+ concentrations. Since effects of Ca2+ are readily observed in intact coupled mitochondria, it follows that Mg2+ concentrations within mitochondria are sub saturating for pyruvate dehydrogenase phosphate phosphatase and hence less than 0.5 mM. PMID- 3036059 TI - Molecular biology and immunology of cytomegalovirus. AB - The application of modern biochemical techniques has led to a rapid improvement in our knowledge of the molecular biology of CMV. Several coding regions of the DNA genome have been identified with certainty and major virus-coded proteins have been given provisional names. The cascade expression of the CMV genome has been shown to be controlled by mechanisms similar to those found in other herpes viruses, together with novel post-transcriptional controls which remain to be defined. The control of CMV replication by the host involves both non-specific and specific defence mechanisms. The induction of natural killer cells and interferon early after CMV infection appears to be the most important aspects of the non-specific host defence against the virus. The cell-mediated immune response, in particular the generation of Tc cells against CMV early antigens, is probably the most important facet of the specific immune defence against CMV. When intact these defence mechanisms appear to be efficient in restricting viral replication; however, when such immunity is compromised, the balance rapidly swings in favour of the virus. As our understanding of the interaction between the host and the virus increases, it may be possible to redress the balance in such cases in favour of the host. PMID- 3036062 TI - Fluoroaluminates mimic guanosine 5'-[gamma-thio]triphosphate in activating the polyphosphoinositide phosphodiesterase of hepatocyte membranes. Role for the guanine nucleotide regulatory protein Gp in signal transduction. AB - Fluoride and guanosine 5'-[gamma-thio]triphosphate (GTP gamma S) both activate the hepatocyte membrane polyphosphoinositide phosphodiesterase (PPI-pde) in a concentration-dependent manner. AlCl3 enhances the fluoride effect, supporting the concept that [A1F4]- is the active species. Analysis of the products of inositol lipid hydrolysis demonstrate that phosphatidylinositol bisphosphate is the major lipid to be hydrolysed. Guanosine 5'-[beta-thio]diphosphate (GDP beta S) is an inhibitor of activation of PPI-pde by both fluoride and GTP gamma S. These observations suggest that the guanine nucleotide regulatory protein (termed Gp) bears a structural resemblance to the well-characterized G-proteins of the adenylate cyclase system and the cyclic GMP phosphodiesterase system in phototransduction. PMID- 3036063 TI - Forskolin refractoriness. Exposure to the diterpene alters guanine nucleotide dependent adenylate cyclase and calcium-uptake activity of cells cultured from the rat aorta. AB - Cells with the morphological properties of endothelial cells were cultured from the rat aorta. The cultured cells accumulated 45Ca2+ from the medium in a manner which was stimulated by forskolin and by 8-bromo-cyclic AMP. Pretreating the cultures for 20 h with forskolin diminished forskolin-dependent Ca2+-uptake activity. Adenylate cyclase activity of cultured cell homogenates was stimulated by guanosine 5'-[beta, gamma-imido]triphosphate (p[NH]ppG) and forskolin, and by isoprenaline in the presence, but not in the absence, of guanine nucleotide. p[NH]ppG increased forskolin sensitivity and caused a leftward shift in the forskolin dose-response curve. Pretreating the cultured cells with forskolin for 20 h, conditions that decreased forskolin-dependent Ca2+ uptake, increased basal and guanine nucleotide-dependent adenylate cyclase activity, but not forskolin dependent activity determined in the absence of p[NH]ppG. Forskolin pretreatment diminished p[NH]ppG's capacity to increase forskolin sensitivity, but did not have a significant effect on either the sensitivity of adenylate cyclase to p[NH]ppG or its responsiveness to isoprenaline. These results suggest that the Ca2+-uptake mechanism is cyclic AMP-dependent and that guanine nucleotides mediated forskolin-dependent cyclic AMP production by the intact cells. In addition, there may be different guanine nucleotide requirements for hormone receptor coupling and forskolin activation. PMID- 3036064 TI - Characterization of a low-relative-molecular-mass prolyl 4-hydroxylase from the green alga Chlamydomonas reinhardii. AB - Prolyl 4-hydroxylase was partially purified and characterized from the unicellular green alga, Chlamydomonas reinhardii. This enzyme differed from all the animal and plant prolyl 4-hydroxylases studied so far in that its Mr was only about 40,000 by gel filtration, being thus less than one-sixth of those determined for the vertebrate and higher-plant enzymes. The algal enzyme did not hydroxylate to any significant extent chick-embryo protocollagen or triple helical (Pro-Pro-Gly)10, whereas a low hydroxylation rate was found with denatured (Pro-Pro-Gly)10. Poly(L-proline), which is an effective inhibitor of the vertebrate enzymes but acts as a substrate for some higher-plant enzymes, was a good substrate. In the absence of poly(L-proline) the enzyme catalysed an uncoupled decarboxylation of 2-oxoglutarate. Studies of the Km values for the co substrates and cofactors and the specificity of the 2-oxoglutarate requirement, as well as inhibition studies with selected 2-oxoglutarate analogues, suggested that the catalytic site of the algal enzyme is similar to, but not identical with, those of the vertebrate enzymes. The existence of distinct similarities was further demonstrated by an inhibition of the algal enzyme activity with a monoclonal antibody to the beta-subunit of human prolyl 4-hydroxylase. The amount of prolyl 4-hydroxylase activity in the algal cells was not altered by signals which recognize the presence or absence of the cell wall, as determined in studies on experimental cell-wall regeneration and wall-less mutants. PMID- 3036066 TI - The identification of a new cyclic nucleotide phosphodiesterase activity in human and guinea-pig cardiac ventricle. Implications for the mechanism of action of selective phosphodiesterase inhibitors. AB - Four cyclic nucleotide phosphodiesterase (PDE) activities were separated from low speed supernatants of homogenates of human cardiac ventricle by DEAE-Sepharose chromatography, and designated PDE I-PDE IV in order of elution with an increasing salt gradient. PDE I was a Ca2+/calmodulin-stimulated activity, and PDE II was an activity with a high Km for cyclic AMP which was stimulated by low concentrations of cyclic GMP. Human ventricle PDE III had Km values of 0.14 microM (cyclic AMP) and 4 microM (cyclic GMP), and showed simple Michaelis-Menten kinetics with both substrates. PDE IV is a previously unrecognized activity in cardiac muscle, the human enzyme having Km values of 2 microM (cyclic AMP) and 50 microM (cyclic GMP). PDE III and PDE IV were not activated by cyclic nucleotides or calmodulin. Four PDE activities were also isolated from guinea-pig ventricle, and had very similar kinetic properties. By gel filtration, the Mr of PDE III was 60,000, and that of PDE IV 45,000. The drug SK&F 94120 selectively and competitively inhibited PDE III with a Ki value of 0.8 microM (human), showing simple hyperbolic inhibition kinetics. Rolipram (Schering ZK 62711) and Ro 20 1724 (Roche), which have previously been reported to inhibit PDE III-like activities strongly, were shown to be weak inhibitors of human and guinea-pig PDE III enzymes (Ki values greater than 25 microM), but potent inhibitors of PDE IV [Ki values 2.4 microM (Rolipram) and 3.1 microM (Ro 20-1724) with human PDE IV]. The inhibition in all cases demonstrated simple hyperbolic competition. These observations suggest that the previously reported complex inhibition of PDE III type activities from cardiac muscle was caused by incomplete separation of the PDE III from other enzymes, particularly PDE IV. PMID- 3036065 TI - Procollagenase activator produced by rabbit uterine cervical fibroblasts. AB - Culture medium from rabbit uterine cervical fibroblasts contained a procollagenase and a neutral proproteinase which acts as a procollagenase activator. These two proenzymes have been purified by a combination of ion exchange, affinity and gel chromatographies. The purified neutral proproteinase showed Mr 60,000 with sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. This neutral proproteinase was activated by trypsin, 4-aminophenylmercuric acetate (APMA) and plasmin, and the active species of the proteinase had Mr 53,000 when activated by APMA; kallikrein and urokinase did not activate this proproteinase. The purified neutral proteinase was inhibited by EDTA, 1,10 phenanthroline and rabbit plasma, but not by serine proteinase inhibitors, suggesting that this proteinase is a metal-dependent proteinase. The purified enzyme could also degrade gelatin, casein, proteoglycan and type IV procollagen. The purified procollagenase had Mr 55,000 and was activated by trypsin, APMA and the active neutral proteinase. These activations were accompanied by decrease in Mr, and the activated species had an Mr which was approx. 10,000 less than that of the procollagenase. In particular, procollagenase activation with neutral proteinase depended on incubation time and proteolytic activity of proteinase. These results indicate that activation of procollagenase by the rabbit uterine neutral proteinase is related to limited proteolysis in the procollagenase molecule. PMID- 3036067 TI - Interaction of acetyl phosphate and carbamyl phosphate with plant phosphoenolpyruvate carboxylase. AB - Acetyl phosphate produced an increase in the maximum velocity (Vmax. for the carboxylation of phosphoenolpyruvate catalysed by phosphoenolpyruvate carboxylase. The limiting Vmax. was 22.2 mumol X min-1 X mg-1 (185% of the value without acetyl phosphate). This compound also decreased the Km for phosphoenolpyruvate to 0.18 mM. The apparent activation constants for acetyl phosphate were 1.6 mM and 0.62 mM in the presence of 0.5 and 4 mM phosphoenolpyruvate respectively. Carbamyl phosphate produced an increase in Vmax. and Km for phosphoenolpyruvate. The variation of Vmax./Km with carbamyl phosphate concentration could be described by a model in which this compound interacts with the carboxylase at two different types of sites: an allosteric activator site(s) and the substrate-binding site(s). Carbamyl phosphate was hydrolysed by the action of phosphoenolpyruvate carboxylase. The hydrolysis produced Pi and NH4+ in a 1:1 relationship. Values of Vmax. and Km were 0.11 +/- 0.01 mumol of Pi X min-1 X mg-1 and 1.4 +/- 0.1 mM, respectively, in the presence of 10 mM-NaHCO3. If HCO3- was not added, these values were 0.075 +/- 0.014 mumol of Pi X min-1 X mg-1 and 0.76 +/- 0.06 mM. Vmax./Km showed no variation between pH 6.5 and 8.5. The reaction required Mg2+; the activation constants were 0.77 and 0.31 mM at pH 6.5 and 8.5 respectively. Presumably, carbamyl phosphate is hydrolysed by phosphoenolpyruvate carboxylase by a reaction the mechanism of which is related to that of the carboxylation of phosphoenolpyruvate. PMID- 3036068 TI - The stimulatory effects of asbestos on NADPH-dependent lipid peroxidation in rat liver microsomes. AB - Lipid peroxidation in rat liver microsomes induced by asbestos fibres, crocidolite and chrysotile, is greatly increased in the presence of NADPH, leading to malondialdehyde levels comparable with those induced by CCl4, a very strong inducer of lipid peroxidation. This synergic effect only occurs during the first minutes and could be explained by an increase or a regeneration of the ferrous active sites of asbestos by NADPH, which in turn could rapidly be prevented by the adsorption of microsomal proteins on the surface of the fibres. It is not inhibited by superoxide dismutase, catalase and mannitol, indicating that oxygen radicals are not involved in the reaction. It is also not inhibited by desferrioxamine, indicating that it is not due to a release of free iron ions in solution from the fibres. Lipid peroxidation in NADPH-supplemented microsomes is also greatly increased upon addition of magnetite. This could be linked to the presence of ferrous ions in this solid iron oxide, since the ferric oxides haematite and goethite are completely inactive. PMID- 3036069 TI - The rapid alteration by tri-iodo-L-thyronine in vivo of both the ADP/O ratio and the apparent H+/O ratio in hypothyroid-rat liver mitochondria. AB - Mitochondria from the livers of thyroidectomized rats have a lowered ADP/O ratio, which can be restored to normal within 15 min after intravenous injection of a near-physiological dose of tri-iodothyronine. Thyroidectomy lowered the measured delta pH, which appears to be compensated by a rise (not statistically significant) in the membrane potential, so that the protonmotive force is unaltered. A simple simulation technique is described for use in estimating H+/O ratios by the oxygen-pulse technique, which circumvents the problem that this ratio can be seriously underestimated because of re-uptake of protons from the bulk phase by the mitochondria before their expulsion is complete. By this procedure the H+/O ratio of hypothyroid mitochondria is shown to be lowered by the same factor as the ADP/O ratio, and both these ratios are very rapidly restored in parallel by hormone administration. Although these findings could be consistent with a proposal that tri-iodothyronine rapidly modulates by some mechanism the efficiency of the respiratory-chain-linked proton pumps, the kinetic properties of the proton exchange suggest that the bulk-phase protons measured may not reflect faithfully those that drive the ATP synthetase. PMID- 3036070 TI - Complete amino acid sequence of the A chain of human complement-classical-pathway enzyme C1r. AB - The amino acid sequence of human C1r A chain was determined, from sequence analysis performed on fragments obtained from C1r autolytic cleavage, cleavage of methionyl bonds, tryptic cleavages at arginine and lysine residues, and cleavages by staphylococcal proteinase. The polypeptide chain has an N-terminal serine residue and contains 446 amino acid residues (Mr 51,200). The sequence data allow chemical characterization of fragments alpha (positions 1-211), beta (positions 212-279) and gamma (positions 280-446) yielded from C1r autolytic cleavage, and identification of the two major cleavage sites generating these fragments. Position 150 of C1r A chain is occupied by a modified amino acid residue that, upon acid hydrolysis, yields erythro-beta-hydroxyaspartic acid, and that is located in a sequence homologous to the beta-hydroxyaspartic acid-containing regions of Factor IX, Factor X, protein C and protein Z. Sequence comparison reveals internal homology between two segments (positions 10-78 and 186-257). Two carbohydrate moieties are attached to the polypeptide chain, both via asparagine residues at positions 108 and 204. Combined with the previously determined sequence of C1r B chain [Arlaud & Gagnon (1983) Biochemistry 22, 1758-1764], these data give the complete sequence of human C1r. PMID- 3036072 TI - Purification of phosphatidylinositol kinase from bovine brain myelin. AB - A membrane-bound phosphatidylinositol (PI) kinase (EC 2.7.1.67) was purified by affinity chromatography from bovine brain myelin. This enzyme activity was solubilized with non-ionic detergent and chromatographed on an anion-exchange column. Further purification was achieved by affinity chromatography on PI covalently coupled to epoxy-activated Sepharose, which was eluted with a combination of PI and detergent. The final step in the purification was by gel filtration on an Ultrogel AcA44 column. This procedure afforded greater than 5500 fold purification of the enzyme from whole brain myelin. The resulting activity exhibited a major silver-stained band on SDS/polyacrylamide-gel electrophoresis with an apparent Mr 45,000. The identity of this band as PI kinase was corroborated by demonstration of enzyme activity in the gel region corresponding to that of the stained protein. The purified enzyme exhibited a non-linear dependence on PI as substrate, with two apparent kinetic components. The lower affinity component exhibited a Km similar to that observed for the phosphorylation of phosphatidylinositol 4-phosphate by the enzyme. PMID- 3036071 TI - Short-term stimulation of Na+-dependent amino acid transport by dibutyryl cyclic AMP in hepatocytes. Characteristics and partial mechanism. AB - The short-term protein-synthesis-independent stimulation of alanine transport in hepatocytes was further investigated. Cyclic AMP increased the Vmax. of alanine transport. Amino acid transport via systems A, ASC and N was stimulated. A good correlation was found between the initial rate of transport and the cell membrane potential as calculated from the distribution of Cl-. Cyclic AMP increased the rate of alanine transport, stimulated Na+/K+ ATPase (Na+/K+-transporting ATPase) activity and caused membrane hyperpolarization. The time courses and cyclic AMP dose-dependencies of all three effects were similar. Ouabain abolished the effect of cyclic AMP on Cl- distribution and on transport of alanine. The effect of cyclic AMP on alanine transport and Cl- distribution was mimicked by the antibiotic nigericin; the effect of nigericin was also abolished by ouabain. It is concluded that the effect of cyclic AMP on transport is mediated via membrane hyperpolarization. It is suggested that the primary action of cyclic AMP is to increase the activity of an electroneutral Na+/K+-exchange system in the liver cell plasma membrane, thus hyperpolarizing the membrane by stimulating the electrogenic Na+/K+ ATPase. PMID- 3036073 TI - Short-term hyperthyroidism modulates adenosine receptors and catalytic activity of adenylate cyclase in adipocytes. AB - The effects of short-term hyperthyroidism in vivo on the status of the components of the fat-cell hormone-sensitive adenylate cyclase were investigated. The number of beta-adrenergic receptors was elevated by about 25% in membranes of fat-cells isolated from hyperthyroid rats as compared with euthyroid rats, but their affinity for radioligand was unchanged. Membranes of hyperthyroid-rat fat-cells displayed less than 65% of the normal complement of receptors for [3H]cyclohexyladenosine. The affinity of the receptors for this ligand was normal. In contrast with the marked increase in the amounts of the alpha-subunits of the guanine nucleotide-binding proteins Gi (Mr 41,000) and Go (Mr 39,000) observed in the hypothyroid state [Malbon, Rapiejko & Mangano (1985) J. Biol. Chem. 260, 2558-2564], the amounts of alpha-Gi, alpha-Go as well as alpha-Gs subunits [Mr 42,000 (major) and 46,000/48,000 (minor)] were not changed by hyperthyroidism. Adenylate cyclase activity in response to forskolin, guanosine 5'-[gamma-thio]triphosphate or isoprenaline, in contrast, was decreased by 30-50% in fat-cell membranes from hyperthyroid rats. Fat-cells isolated from hyperthyroid rats accumulated cyclic AMP to less than 50% of the extent in their euthyroid counterparts in the presence of adenosine deaminase and either adrenaline or forskolin, suggesting a decrease in the amount or activity of the catalytic subunit of adenylate cyclase. In the absence of exogenous adenosine deaminase, cyclic AMP accumulation in response to adrenaline was elevated rather than decreased in fat-cells from hyperthyroid rats. The inhibitory influence of adenosine is apparently limited in the hyperthyroid state by the decreased complement of inhibitory R-site purinergic receptors in these fat-cells. Short term hyperthyroidism modulates the fat-cell adenylate cyclase system at the receptor level (beta-receptor number increased, R-site purinergic-receptor number decreased) and the catalytic subunit of adenylate cyclase. PMID- 3036075 TI - A putative protein-sequestration site involving intermediate filaments for protein degradation by autophagy. Studies with transplanted Sendai-viral envelope proteins in HTC cells. AB - Reconstituted Sendai-viral envelopes (RSVE) were fused with hepatoma tissue culture (HTC) cells, thereby introducing viral membrane glycoproteins into the plasma membrane [Earl, Billett, Hunneyball & Mayer (1987) Biochem. J. 241, 801 807]. Fractionation of homogenized cells on Nycodenz gradients shows that much of the viral 125I-labelled HN and F proteins were rapidly sequestered into a dense fraction distinct from fractions containing plasma membrane, lysosomes and mitochondria. Electron microscopy (results not shown) indicates that the dense fraction contains nuclear residues, multivesicular structures, dense bodies and fibrous structures. Both the dense fraction and a hexosaminidase-enriched fraction contain trichloroacetic acid-insoluble radioactivity, including intact 125I-labelled viral proteins. The viral proteins are progressively transferred from the dense fraction to the hexosaminidase-enriched fraction; the transfer is retarded by 50 micrograms of leupeptin/ml. Trichloroacetic acid-soluble radiolabel is progressively released into the culture medium as the proteins are degraded. Within 5 h after transplantation of viral HN and F proteins into recipient cells, a proportion (approx. 45%) of the 125I-labelled glycoproteins cannot be extracted by sequentially treating cells with digitonin (1 mg/ml), Triton X-100 (1%, w/v) and 0.3 M-KI. HN and F proteins in the non-extractable residue are tightly associated with nuclear-intermediate-filament (vimentin) material, as shown by Western blots and electron microscopy. The viral proteins are progressively transferred out of the nuclear-intermediate-filament residue; the transfer is slowed when cells are cultured with leupeptin. The data are consistent with the notion that transplanted viral HN and F proteins are sequestered to a perinuclear site in tight association with intermediate filaments before transfer into the autophagolysosomal system for degradation. PMID- 3036074 TI - Sendai-viral HN and F glycoproteins as probes of plasma-membrane protein catabolism in HTC cells. Studies with fusogenic reconstituted Sendai-viral envelopes. AB - Reconstituted Sendai-viral envelopes (RSVE) were produced by the method of Vainstein, Hershkovitz, Israel & Loyter [(1984) Biochim. Biophys. Acta 773, 181 188]. RSVE are fusogenic unilamellar vesicles containing two transmembrane glycoproteins: the HN (haemagglutinin-neuraminidase) protein and the F (fusion) factor. The fate of the viral proteins after fusion-mediated transplantation of RSVE into hepatoma (HTC) cell plasma membranes was studied to probe plasma membrane protein degradation. Both protein species are degraded at similar, relatively slow, rates (t1/2 = 67 h) in HTC cells fused with RSVE in suspension. Even slower degradation rates for HN and F proteins (t1/2 = 93 h) were measured when RSVE were fused with HTC cells in monolayer. Lysosomal degradation of the transplanted viral proteins is strongly implicated by the finding that degradation of HN and F proteins is sensitive to inhibition by 10 mM-NH4Cl (81%) and by 50 micrograms of leupeptin/ml (70%). PMID- 3036076 TI - Mung bean (Phaseolus aureus) nuclease. A mechanistic investigation of the DNA cleavage reaction using a dinucleoside phosphorothioate. AB - Mung-bean (Phaseolus aureus) nuclease has been found to cleave the Sp diastereoisomer of 5'-O-thymidyl 3'-O-(2'-deoxyadenosyl)phosphorothioate, (Sp) d[Ap(S)T], in 18O-labelled water with inversion of configuration at phosphorus to give (Sp)-thymidine 5'-[16O, 18O]phosphorothioate, the stereochemistry of which was deduced by methylation to (Rp,Sp)-thymidine 5'-S-methyl-O-methyl [16O,18O]phosphorothioate and 31P-n.m.r. analysis. This result is consistent with a mechanism involving a direct 'in-line' attack of water on DNA for the nuclease catalysed reaction without the involvement of a covalent nucleotidylated-enzyme intermediate. PMID- 3036078 TI - Sustained oscillations in extracellular calcium concentrations upon hormonal stimulation of perfused rat liver. AB - Sustained oscillations in extracellular free Ca2+ were shown to occur on addition of vasopressin, phenylephrine or angiotensin II in isolated rat liver perfused with low (10 microM)-Ca2+ medium. The amplitude and frequency of oscillation depend on hormone concentration. In contrast, Ca2+ releases on addition of ATP, t butyl hydroperoxide or arachidonate do not exhibit oscillatory behaviour. The vasopressin-induced oscillations were suppressed by glucagon and dibutyryl 3',5' cyclic AMP, but not by dibutyryl 3',5'-cyclic GMP. These observations in the extracellular space complement observations by Woods, Cuthbertson & Cobbold [(1986) Nature (London) 319, 600-602] on oscillations in intracellular free Ca2+ in single liver cells. PMID- 3036077 TI - Effects of glucagon and Ca2+ on the metabolism of phosphatidylinositol 4 phosphate and phosphatidylinositol 4,5-bisphosphate in isolated rat hepatocytes and plasma membranes. AB - Rat hepatocytes whose phosphatidylinositol 4-phosphate (PIP) and phosphatidylinositol 4,5-bisphosphate (PIP2) had been labelled for 60 min with 32P were treated with glucagon for 10 min or phenylephrine for 2 min. Glucagon caused a 20% increase in PIP but no change in PIP2 whereas phenylephrine caused a similar increase in PIP but a 15% decrease in PIP2. Addition of both hormones together for 10 min produced a 40% increase in PIP. A crude liver mitochondrial fraction incubated with [32P]Pi and ADP incorporated label into PIP, PIP2 and phosphatidic acid. The PIP2 was shown to be in contaminating plasma membranes and PIP in both lysosomal and plasma-membrane contamination. A minor but definitely mitochondrial phospholipid, more polar than PIP2, was shown to be labelled with 32P both in vitro and in hepatocytes. The rate of 32P incorporation into PIP was faster in mitochondrial/plasma-membrane preparations from rats treated with glucagon or if 3 microM-Ca2+ and Ruthenium Red were present in the incubation buffer. Loss of 32P from membranes labelled in vitro was shown to be accompanied by formation of inositol 1,4,5-trisphosphate (IP3) and inositol 1,4-bisphosphate, and was faster in preparations from glucagon-treated rats or in the presence of 3 microM-Ca2+. It is concluded that glucagon stimulates both PIP2 phosphodiesterase and phosphatidylinositol kinase activities, as does the presence of 3 microM Ca2+. The resulting formation of IP3 may be responsible for the observed release of intracellular Ca2+ stores. The roles of a guanine nucleotide regulatory protein and phosphorylation in mediating these effects are discussed. PMID- 3036079 TI - The inhibition by diphenyleneiodonium and its analogues of superoxide generation by macrophages. AB - Peritoneal macrophages were elicited in rats by using casein as a stimulus; when stimulated with phorbol 12-myristate 13-acetate (PMA) they produced O2.-. Nearly 60% of the total cytochrome b had a low Em,7.0 of -247 mV, typical of the cytochrome b component found in the NADPH-dependent O2(.-)-generating oxidase of neutrophils. The rate of O2.- generation by macrophages was 1.23 mol of O2.-/s per mol of cytochrome b. Treatment of intact macrophages with diphenyleniodonium (DPI) at 0.9 microM caused 50% inhibition of PMA-induced O2.- generation, with little effect on mitochondrial respiratory activity; KCN inhibited respiratory activity without affecting PMA-induced O2.- generation. A similar specificity of inhibition was found for di-2-thienyliodonium (50% inhibition of O2.- generation at 0.5 microM) and, at higher concentrations, for diphenyl iodonium. When macrophage suspensions were incubated with [125I]DPI followed by autoradiography of SDS/polyacrylamide-gel-electrophoresis-separated polypeptides, radioactivity was most strongly associated with a band of Mr 45,000, similar to that found in neutrophils [Cross & Jones (1986) Biochem. J. 237, 111-116]. The O2(.-) generating oxidase of macrophages appears to have components in common with the NADPH oxidase of neutrophils, despite differences in activity. Its sensitivity to DPI suggests that selective prevention of radical generation by macrophages in vivo is possible. PMID- 3036080 TI - Thrombin-induced phosphoinositide hydrolysis in platelets. Receptor occupancy and desensitization. AB - The relationship between occupancy of thrombin receptors on platelets and enhanced phosphoinositide hydrolysis was analysed by examination of the dose response relationship, the effects of thrombin inhibitors and the contribution of secondary effects. Washed human platelets were labelled with [3H]inositol, and agonist-induced accumulation of labelled inositol phosphates was measured. The dose-response curves and the time courses for alpha-thrombin- or gamma-thrombin induced accumulation of inositol phosphates were similar to those for dense granule secretion. Addition of the thrombin inhibitor hirudin to thrombin activated platelets revealed that the continuous presence of active thrombin was required to maintain the accumulation of labelled inositol phosphates; the total production of inositol phosphates increased with longer periods of exposure to thrombin, reaching a maximum between 5 and 10 min. After activation with thrombin, the ability of a second, greater, addition of thrombin to induce additional phosphoinositide hydrolysis decreased with time; it was absent within 10 min after the first addition. The failure to sustain accumulation of labelled inositol phosphates or to respond to a second addition of thrombin beyond 10 min was not due to depletion of the pool of labelled precursors, because the platelets retained their ability to respond to collagen. Addition of ADP consuming enzymes decreased sensitivity to thrombin, but inhibition of cyclo oxygenase with indomethacin did not impair the thrombin-induced hydrolysis of phosphoinositides. It was concluded that thrombin-induced hydrolysis of phosphoinositides has characteristics consistent with mediation by a receptor that is similar to that that triggers dense-granule secretion, requires continuous presence of active thrombin to be maintained, is mediated by a receptor that displays thrombin-induced desensitization, and is only partially enhanced by secondary agents. PMID- 3036081 TI - Time-dependent inactivation of chick-embryo prolyl 4-hydroxylase by coumalic acid. Evidence for a syncatalytic mechanism. AB - From the structure-activity relationships of known competitive inhibitors, coumalic acid (2-oxo-1,2H-pyran-5-carboxylic acid) was deduced to be a potential syncatalytic inhibitor for chick-embryo prolyl 4-hydroxylase. The compound caused time-dependent inactivation, the reaction rate being first-order. The inactivation constant was 0.094 min-1, the Ki 17 mM and the bimolecular rate constant 0.09 M-1 X S-1. Human prolyl 4-hydroxylase and chick embryo lysyl hydroxylase were also inactivated, though to a lesser extent. Inactivation could be prevented by adding high concentrations of 2-oxoglutarate or its competitive analogues to the reaction mixture. In Lineweaver-Burk kinetics, coumalic acid displayed S-parabolic competitive inhibition with respect to 2-oxoglutarate. The inactivation reaction had cofactor requirements similar to those for the decarboxylation of 2-oxoglutarate. Enzymic activity was partially preserved in the absence of iron, but the rescue was incomplete, owing to decreased stability of the enzyme under this condition. Coumalic acid also decreased the electrophoretic mobility of the alpha-subunit, but the beta-subunit was not affected. Prolonged incubation of coumalic acid above pH 6.8 led to loss of its inactivating potency, owing to hydrolysis. It is concluded that the inactivation of prolyl 4-hydroxylase by coumalic acid is due to a syncatalytic mechanism. The data also suggest that the 2-oxoglutarate-binding site of the enzyme is located within the alpha-subunit. PMID- 3036084 TI - Synthesis of mucous glycoproteins by rabbit tracheal cells in vitro. Modulation by substratum, retinoids and cyclic AMP. AB - One function of airway epithelium is the secretion of mucins, which comprise an important component of the mucous lining layer. We demonstrate that rabbit tracheal epithelial cells grown in primary culture incorporate [3H]glucosamine into material released into the medium which is characterized as mucin by the following criteria: high Mr, monosaccharide composition, ion-exchange behaviour different from that of glycosaminoglycans and oligosaccharides attached via N acetylgalactosamine. The production of mucin by the cells requires growth on a substratum of collagen gel and is enhanced by retinoids in the extracellular medium. In the presence of retinoids, 8-bromo cyclic AMP and factors present in medium from 3T3 fibroblasts each further stimulate mucin production. These results indicate that an isolated epithelial-cell culture system, in the absence of nervous, mesenchymal or other tissue types, can be used to answer questions about the regulation of mucin production at the cellular level. PMID- 3036082 TI - The acylation of lysophosphoradylglycerocholines in guinea-pig heart mitochondria. AB - The importance of the deacylation-reacylation pathway for attaining the desired fatty acid composition in microsomal phospholipids has been well established. It is not clear, however, whether this mechanism is of equal importance in mitochondria. The absence of acyltransferase activity in mammalian heart mitochondria has been reported in a number of studies. In the present study we report the presence of acyltransferase activities for lysophosphoradylglycerocholines in guinea-pig heart mitochondria. This enzyme showed properties that were considerably different from those of the microsomal enzymes. Of all the acyl-CoAs tested (C18:0, C18:1, C18:2 and C20:4) the mitochondrial enzyme utilized only linoleoyl-CoA as fatty acyl donor and utilized both 1-acyl-sn-glycero-3-phosphocholine and 1-alkenyl-sn-glycero-3-phosphocholine as fatty acyl acceptors. The presence of significant quantities of fatty acids other than linoleate at the C-2 position of mitochondrial acylglycerophosphocholines, coupled with the specificity of the enzyme for linoleoyl-CoA, suggest that, in addition to reacylation, other mechanisms play a significant role in producing the molecular composition of these phospholipids found in the mitochondria. PMID- 3036083 TI - The vasoactive intestinal peptide receptor on intact human colonic adenocarcinoma cells (HT29-D4). Evidence for its glycoprotein nature. AB - We have previously shown that the mono [125I]iodinated vasoactive intestinal peptide (125I-VIP) could be covalently cross-linked on intact colonic adenocarcinoma cells (HT29). A major Mr 67,000 and a minor Mr 120,000 cross linked polypeptides have been characterized [Muller, Luis, Fantini, Abadie, Giannellini, Marvaldi & Pichon (1985) Eur. J. Biochem. 151, 411-417]. The glycoprotein nature of these species was investigated using endo-beta acetylglucosaminidase F (Endo F) treatment, enzymic and chemical desialylation and wheat germ agglutinin (WGA)-Sepharose affinity chromatography. Affinity labelled VIP-binding proteins solubilized by Nonidet P-40 bound to WGA-Sepharose and could be eluted specifically with N-acetyl-D-glucosamine. Treatment with Endo F resulted in an increased electrophoretic mobility of both polypeptides. The major and the minor VIP-binding proteins were converted respectively into Mr 47,000 and 100,000 species, indicating removal of 20 kDa of N-linked oligosaccharides. Deglycosylation with trifluoromethanesulphonic acid also led to a 20 kDa loss in mass of the Mr 67,000 component, indicating the absence of additional O-linked sugars on this polypeptide. The presence of sialic acid on the major VIP-binding protein was demonstrated after treatment of intact cells with neuraminidase or by chemical desialylation with hydrochloric acid. We conclude from this study that the VIP receptor from intact HT29-D4 cells is a glycoprotein with N-linked oligosaccharide side chains containing sialic acid. PMID- 3036085 TI - Enzymic dephosphorylation of D-myo-inositol 1,4-bisphosphate in rat brain. AB - Inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] and inositol 1,4-bisphosphate [Ins(1,4)P2] phosphatase activities were measured in both 180,000 g (60 min) particulate and supernatant fractions of rat brain homogenates. Although Ins(1,4,5)P3 was mostly hydrolysed by a particulate phosphatase [Erneux, Delvaux, Moreau & Dumont (1986) Biochem. Biophys. Res. Commun. 134, 351-358], Ins(1,4)P2 phosphatase was predominantly soluble. The latter enzyme was Mg2+-dependent and sensitive to thiol-blocking agents (e.g. p-hydroxymercuribenzoate). In contrast with Ins(1,4,5)P3 phosphatase activity measured in the soluble fraction, Ins(1,4)P2 phosphatase was insensitive to 0.001-1 mM-2,3-bisphosphoglycerate. Lithium salts, widely used in psychiatric treatment, inhibited both Ins(1,4)P2 and Ins(1)P1 phosphatase activities of the crude soluble fraction. In particular, 50% inhibition of phosphatase activity, with 2 microM-Ins(1,4)P2 as substrate, was achieved at 3-5 mM-LiCl. At these concentrations, LiCl did not change Ins(1,4,5)P3 phosphatase activity measured in the same fraction with 1-4 microM Ins(1,4,5)P3 as substrate. Chromatography of the soluble fraction of a rat brain homogenate on DEAE-cellulose resolved three phosphatase activities. These forms, peaks I, II and III, dephosphorylated Ins(1,4,5)P3, Ins(1)P1 and Ins(1,4)P2 respectively. If LiCl (10 mM) was included in the assay mixture, it inhibited both peak-II Ins(1)P1 phosphatase and peak-III Ins(1,4)P2 phosphatase, suggesting the existence of at least two Li+-sensitive phosphatases. PMID- 3036086 TI - Mouse and human ornithine decarboxylase genes. Methylation polymorphism and amplification. AB - With the use of the isoschizomeric restriction endonucleases HpaII and MspI, we found that mouse tumour ornithine decarboxylase (ODC; EC 4.1.1.17) genes are extensively methylated. ODC genes in L1210 mouse leukaemia cells were apparently more methylated than in Ehrlich ascites carcinoma, as revealed by the use of HpaII endonuclease, yet the digestion of genomic DNA isolated from these two murine tumour cell lines with MspI, which cleaves at a CCGG sequence, also with internally methylated cytosine, resulted in an apparently identical restriction pattern. It is possible that the amplification of ODC genes in Ehrlich ascites carcinoma cells in response to 2-difluoromethylornithine (DFMO) was associated with hypomethylation, or that less-methylated genes were amplified. A human myeloma (Sultan) cell line only revealed three separate hybridization signals when cleaved with HpaII. One of these signals was amplified under the pressure of DFMO. When cleaved with MspI, these three HpaII fragments disappeared and were replaced by a double signal of 2.3-2.4 kilobase-pairs (kbp) in size. The amplified ODC sequences in the Sultan myeloma cell line apparently originated from chromosome 2, as indicated by a unique hybridization signal in a 5.8 kbp HindIII fragment specific for the human ODC locus on chromosome 2. A comparison of different human cells, the Sultan myeloma, a lymphocytic B-cell leukaemia (Ball), normal mononuclear leucocytes and leucocytes obtained from leukaemia patients, revealed interesting differences in the methylation of ODC genes. The use of two restriction endonucleases (HpaII and CfoI), the cleavage site for both of which contains a CG sequence and which only cleave when cytosine is unmethylated, indicated that ODC genes in the lymphocytic leukaemia cells were much less methylated than those in the normal leucocytes or in the Sultan cells. PMID- 3036087 TI - The insulin- and glucagon-stimulated 'dense-vesicle' high-affinity cyclic AMP phosphodiesterase from rat liver. Purification, characterization and inhibitor sensitivity. AB - The hormone-stimulated 'dense-vesicle' cyclic AMP phosphodiesterase was solubilized as a proteolytically 'clipped' species, and purified to apparent homogeneity from rat liver with a 2000-3000-fold purification and a 13-18% yield. It appeared to be a dimer (Mr 112,000), of two Mr-57,000 subunits. Solubilization of either a liver or a hepatocyte membrane fraction, with sodium cholate in the presence of the protein inhibitor benzamidine, identified three protein bands which could be immunoprecipitated by a polyclonal antibody raised against the pure enzyme. The major band at Mr 62,000 is suggested to be the native 'dense vesicle' enzyme, having a Mr-5000 extension which serves to anchor this enzyme to the membrane and which is cleaved off during proteolytic solubilization; the Mr 200,000 band is an aggregate of the Mr-62,000 species, and the Mr-63,000 species is possibly a precursor. The purified 'clipped' enzyme hydrolysed cyclic AMP with kinetics indicative of apparent negative co-operativity, with a Hill coefficient (h) of 0.43 and limiting kinetic constants of Km1 = 0.3 +/- 0.05 microM, Km2 = 29 +/- 6 microM, Vmax.1 = 0.114 +/- 0.015 unit/mg of protein and Vmax.2 = 0.633 +/- 0.054 unit/mg of protein. It hydrolysed cyclic GMP with Michaelis kinetics, Km = 10 +/- 1 microM and Vmax. = 4.1 +/- 0.2 units/mg of protein. Cyclic GMP was a potent inhibitor of cyclic AMP hydrolysis, with an IC50 (concn. giving 50% inhibition) of 0.20 +/- 0.01 microM-cyclic GMP when assayed at 0.1 microM-cyclic AMP. This enzyme was inhibited potently by several drugs known to exert positive inotropic effects on the heart, was extremely thermolabile, with a half-life of 4.5 +/- 0.5 min at 40 degrees C, and was shown to be distinct from the rat liver insulin-stimulated peripheral-plasma-membrane cyclic AMP phosphodiesterase [Marchmont, Ayad & Houslay (1981) Biochem. J. 195, 645-652]. PMID- 3036088 TI - Dephosphorylation of myo-inositol 1,4,5-trisphosphate and myo-inositol 1,3,4 triphosphate. AB - We have augmented our previous studies [Storey, Shears, Kirk & Michell (1984) Nature (London) 312, 374-376] on the subcellular location and properties of Ins(1,4,5)P3 (inositol 1,4,5-trisphosphate) phosphatases in rat liver and human erythrocytes. We also investigate Ins(1,3,4)P3 (inositol 1,3,4-trisphosphate) metabolism by rat liver. Membrane-bound and cytosolic Ins(1,4,5)P3 phosphatases both attack the 5-phosphate. The membrane-bound enzyme is located on the inner face of the plasma membrane, and there is little or no activity associated with Golgi apparatus. Cytosolic Ins(1,4,5)P3 5-phosphatase (Mr 77,000) was separated by gel filtration from Ins(1,4)P2 (inositol 1,4-bisphosphate) and inositol 1 phosphate phosphatases (Mr 54,000). Ins(1,4,5)P3 5-phosphatase activity in hepatocytes was unaffected by treatment of the cells with insulin, vasopressin, glucagon or dibutyryl cyclic AMP. Ins(1,4,5)P3 5-phosphatase activity in cell homogenates was unaffected by changes in [Ca2+] from 0.1 to 2 microM. After centrifugation of a liver homogenate at 100,000 g, Ins(1,3,4)P3 phosphatase activity was largely confined to the supernatant. The sum of the activities in the supernatant and the pellet exceeded that in the original homogenate. When these fractions were recombined, Ins(1,3,4)P3 phosphatase activity was restored to that observed in unfractionated homogenate. Ins(1,3,4)P3 was produced from Ins(1,3,4,5)P4 (inositol 1,3,4,5-tetrakisphosphate) and was metabolized to a novel InsP2 that was the 3,4-isomer. Ins(1,3,4)P3 phosphatase activity was not changed by 50 mM-Li+ or 0.07 mM-Ins(1,4)P2 alone, but when added together these agents inhibited Ins(1,3,4)P3 metabolism. In Li+-treated and vasopressin stimulated hepatocytes, Ins(1,4)P2 may reach concentrations sufficient to inhibit Ins(1,3,4)P3 metabolism, with little effect on Ins(1,4,5)P3 hydrolysis. PMID- 3036089 TI - Internalization of insulin receptors and HLA antigens in human hepatoma cells. AB - Human HepG2 hepatoma cells express a high number of insulin receptors. Growing cells exhibit 70% of their insulin receptors on the plasma membrane. Moreover, cell-surface insulin receptors form molecular complexes with class I major histocompatibility antigens, as determined by co-immunoprecipitation of the receptors by anti-class I monoclonal antibodies. On exposure to saturating concentrations of insulin, the hormone is rapidly internalized into a Pronase resistant compartment. Internalization of insulin is accompanied by a rapid (t1/2 = 2-3 min) redistribution of insulin receptors from the cell surface to an intracellular compartment. On removal of insulin from the medium, functional receptors recycle back to the plasma membrane, where they can bind insulin again. With chronic exposure of HepG2 cells to insulin, the initial redistribution of receptors is followed by a slow (t1/2 = 9 h) down-regulation of the receptors. Finally, notwithstanding their interaction at the cell surface, insulin receptors and class I major histocompatibility antigens are internalized at different rates and with independent regulation. PMID- 3036090 TI - A new procedure for the purification of monodisperse highly active cytochrome c oxidase from bovine heart. AB - A simple and rapid method for the isolation of a large quantity of cytochrome c oxidase from bovine heart mitochondria was developed, based on selective solubilization of mitochondrial protein with first Triton and then lauryl maltoside. Gel filtration shows that the lauryl maltoside-solubilized oxidase preparation is in a hydrodynamically homogeneous state with a Stokes radius of 7.5 +/- 0.2 nm. It contains 8.0 mumol of haem (with an a/a3 ratio of 1)/g of protein. The catalytic constant (maximum turnover number) with respect to cytochrome c approaches 600 S-1. After further purification of the solubilized enzyme on a sucrose-gradient centrifugation, the purified enzyme has a haem content of 10.3 mumol/g of protein and eight major polypeptide bands shown on SDS/polyacrylamide-gel electrophoresis. PMID- 3036091 TI - Regulation of ornithine decarboxylase activity by spermidine and the spermidine analogue N1N8-bis(ethyl)spermidine. AB - Polyamine biosynthesis in intact cells can be exquisitely controlled with exogenous polyamines through the regulation of rate-limiting biosynthetic enzymes, particularly ornithine decarboxylase (ODC). In an attempt to exploit this phenomenon as an antiproliferative strategy, certain polyamine analogues have been identified [Porter, Cavanaugh, Stolowich, Ganis, Kelly & Bergeron (1985) Cancer Res. 45, 2050-2057] which lower ODC activity in intact cells, have no direct inhibitory effects on ODC, are incapable of substituting for spermidine (SPD) in supporting cell growth, and are growth-inhibitory at micromolar concentrations. In the present study, the most effective of these analogues, N1N8 bis(ethyl)SPD (BES), is compared with SPD in its ability to regulate ODC activity in intact L1210 cells and in the mechanism(s) by which this is accomplished. With respect to time and dose-dependence of ODC suppression, both polyamines closely paralleled one another in their response curves, although BES was slightly less effective than SPD. Conditions of minimal treatment leading to near-maximal ODC suppression (70-80%) were determined and found to be 3 microM for 2 h with either SPD or BES. After such treatment, ODC activity was fully recovered within 2-4 h when cells were re-seeded in drug-free media. By assessing BES or [3H]SPD concentrations in treated and recovered cells, it was possible to deduce that an intracellular accumulation of BES or SPD equivalent to less than 6.5% of the combined cellular polyamine pool was sufficient to invoke ODC regulatory mechanisms. Decreases in ODC activity after BES or SPD treatment were closely paralleled by concomitant decreases in ODC protein. Since cellular ODC mRNA was not similarly decreased by either BES or SPD, it was concluded that translational and/or post-translational mechanisms, such as increased degradation of ODC protein or decreased translation of ODC mRNA, were probably responsible for regulation of enzyme activity. Experimental evidence indicated that neither of these mechanisms seemed to be mediated by cyclic AMP or ODC-antizyme induction. On the basis of the consistent similarities between BES and SPD in all parameters studied, it is concluded that the analogue most probably acts by the same mechanisms as SPD in regulating polyamine biosynthesis. PMID- 3036093 TI - Galactoside-proton symport in a lacYUN mutant of Escherichia coli investigated by analysis of transport progress curves. AB - The kinetics of galactoside-proton symport catalysed by a wild-type strain and one carrying a mutation, previously reported to cause uncoupling of the symport reaction, have been examined. The mutation does not affect the stoichiometry during the initial period of uptake, when the internal concentration of galactoside is low, but it does result in much greater competition from the galactoside as it is accumulated. Simple methods for the analysis of the uptake progress curves have been developed and used to estimate the initial rate of uptake and affinity for internal galactoside. The maximum rate of uptake is decreased by a factor of 2 at most whereas the affinity for internal galactoside is increased up to 50-fold by the mutation. The pH-dependence of the galactoside efflux reaction is changed in a manner which suggests that the defect is in the interaction between proton-binding and galactoside-binding sites rather than in the structure of either site. PMID- 3036092 TI - Evidence that inositol 1-phosphate in brain of lithium-treated rats results mainly from phosphatidylinositol metabolism. AB - In cerebral cortex of rats treated with increasing doses of LiCl, the relative concentrations of Ins(1)P, Ins(4)P and Ins(5)P (when InsP is a myo-inositol phosphate) are approx. 10:1:0.2 at all doses. In rats treated with LiCl followed by increasing doses of pilocarpine a similar relationship occurs. myo-Inositol-1 phosphatase (InsP1ase) from bovine brain hydrolyses Ins(1)P, Ins(4)P and Ins(5)P at comparable rates, and these substrates have similar Km values. The hydrolysis of Ins(4)P is inhibited by Li+ to a greater degree than is hydrolysis of Ins(1)P and Ins(5)P. D-Ins(1,4,5)P3 and D-Ins(1,4)P2 are neither substrates nor inhibitors of InsP1ase. A dialysed high-speed supernatant of rat brain showed a greater rate of hydrolysis of Ins(1)P than of D-Ins(1,4)P2 and a lower sensitivity of the bisphosphate hydrolysis to LiCl, as compared with the monophosphate. That enzyme preparation produced Ins(4)P at a greater rate than Ins(1)P when D-Ins(1,4)P2 was the substrate. The amount of D-Ins(3)P [i.e. L Ins(1)P, possibly from D-Ins(1,3,4)P3] is only 11% of that of D-Ins(1)P on stimulation with pilocarpine in the presence of Li+. DL-Ins(1,4)P2 was hydrolysed by InsP1ase to the extent of about 50%; both Ins(4)P and Ins(1)P are products, the former being produced more rapidly than the latter; apparently L-Ins(1,4)P2 is a substrate for InsP1ase. Li+, but not Ins(2)P, inhibited the hydrolysis of L Ins(1,4)P2. The following were neither substrates nor inhibitors of InsP1ase; Ins(1,6)P2, Ins(1,2)P2, Ins(1,2,5,6)P4, Ins(1,2,4,5,6)P5, Ins(1,3,4,5,6)P5 and phytic acid. myo-Inositol 1,2-cyclic phosphate was neither substrate nor inhibitor of InsP1ase. We conclude that the 10-fold greater tissue contents of Ins(1)P relative to Ins(4)P in both stimulated and non-stimulated rat brain in vivo are the consequence of a much larger amount of PtdIns metabolism than polyphosphoinositide metabolism under these conditions. PMID- 3036094 TI - The mechanism of potentiation of horseradish peroxidase-catalysed oxidation of NADPH by porphyrins. AB - Several porphyrins, including HpD (haematoporphyrin derivative), potentiate the oxidation of NADPH by horseradish peroxidase/H2O2. To elucidate the mechanism of potentiation, the following observations are relevant. During peroxidase catalysed NADPH oxidation, O2-.(superoxide radical) is generated, as judged from superoxide dismutase-inhibitable cytochrome c reduction. This generation of O2-. is suppressed by HpD. Peroxidase-catalysed NADPH oxidation is stimulated by superoxide dismutase and by anaerobic conditions. Under anaerobic conditions HpD has no influence on peroxide-catalysed NADPH oxidation. Previous studies have shown that horseradish peroxidase is inhibited by O2-.. Thus the experimental results indicate that the potentiating effect of HpD can be explained by its ability to inhibit O2-. generation in the horseradish peroxidase/H2O2/NADPH system. PMID- 3036095 TI - Characterization of glucose transport in an insulin-secreting cell line. AB - The rat insulinoma-derived RINm5F cell line retains many differentiated functions of islet beta-cells. However, it fails to recognize glucose as an insulin secretagogue in the physiological concentration range. With this cell line, glucose-transport kinetics were investigated, by using a double-label technique with the non-metabolizable glucose analogue 3-O-methylglucose (OMG). RINm5F cells possess a passive glucose-transport system with high capacity and low affinity. Equilibration across the plasma membrane of extracellular OMG concentrations up to at least 20 mM is achieved within 2 min at 37 degrees C. The half-saturation of OMG uptake occurs at 32 mM. At lower temperatures OMG uptake is markedly retarded, with a temperature coefficient (Q10) of 2.9. As indicated by efflux measurements, transport is symmetrical. Cytochalasin B at micromolar concentrations and phlorrhizin in millimolar concentrations are potent inhibitors of OMG uptake. Neutralization of the secreted insulin with antibodies does not alter OMG uptake kinetics. The glucose metabolism of RINm5F cells is much exaggerated compared with that of islet beta-cells. Nonetheless, when measured in parallel to uptake, transport exceeds by far the rate of metabolism at glucose concentrations above 3 mM. Measurements of intracellular D-glucose reveal a lower intracellular glucose concentration relative to the extracellular in RINm5F cells. This seems to be due to abnormalities in the subsequent steps of glucose metabolism, rather than to abnormalities in hexose uptake. The loss of glucose induced insulin release in RINm5F cells cannot be explained by alterations in hexose transport. PMID- 3036096 TI - Activation of adenylate cyclase in human platelet membranes by guanosine 5'-[beta gamma-imido]triphosphate is inhibited by cyclic-AMP-dependent phosphorylation. Slow activation occurs in the absence of ATP. AB - Incubation of platelet membranes with guanosine 5'-[beta gamma-imido]triphosphate causes a slow increase in GS (stimulatory GTP-binding protein) activation of adenylate cyclase. Mg2+ is necessary for this slow activation. This process is inhibited in the presence of ATP, and inhibition is greater if cyclic AMP is also included in the incubation. Adenosine 5'-[beta gamma-imido]triphosphate instead of ATP in the incubation facilitates the slow activation in the presence of cyclic AMP, and incubation of membranes with cyclic-AMP-dependent protein kinase inhibitor decreased inhibition of the slow activation of adenylate cyclase by ATP and cyclic AMP. A protein of 45 kDa in platelet membranes is phosphorylated in a cyclic-AMP-dependent manner. The transition from a reversibly activated form of GS to an irreversibly activated form is substantially slower in the presence of ATP and cyclic AMP. We propose that guanosine 5'-[beta gamma-imido]triphosphate activated GS may exist in phosphorylated or non-phosphorylated forms, and that the non-phosphorylated form is the more active of the two species. The non phosphorylated form of GS may correspond to the irreversibly activated state. PMID- 3036097 TI - The role of insulin in the modulation of glucagon-dependent control of phenylalanine hydroxylation in isolated liver cells. AB - The stimulation of phenylalanine hydroxylation in isolated liver cells by sub maximally effective concentrations of glucagon (less than 0.1 microM) is antagonized by insulin (0.1 nM-0.1 microM). This phenomenon is a consequence of a decrease in the glucagon-stimulated phosphorylation of phenylalanine hydroxylase from liver cells incubated in the presence of insulin. The impact of insulin on the phosphorylation state and activity of the hydroxylase is mimicked by incubation of liver cells in the presence of orthovanadate (10 microM). A series of cyclic AMP and cyclic GMP analogues enhanced phenylalanine hydroxylation: in each case insulin diminished the stimulation of flux. These results are discussed in the light of the characteristics of insulin action on other metabolic processes. PMID- 3036098 TI - Peroxidase and peroxidase-oxidase activities of isolated human myeloperoxidases. AB - Isolated neutrophils from healthy donors were used for the isolation of four highly purified forms of myeloperoxidase as determined by spectral (A430/A280 ratio 0.80-0.87) and enzyme-activity measurements. Although the myeloperoxidases exhibited different elution profiles on cation-exchange chromatography, gel filtration indicated similar relative molecular masses. When these forms were assayed for peroxidase and peroxidase-oxidase activities with several substrates, they all exhibited virtually the same specific activities. These results suggest that possible functional differences between the enzymes may be related to differences in their sites of action rather than to differences in enzyme activity. Myeloperoxidase from a patient with chronic myeloid leukaemia also revealed a similar heterogeneity on cation-exchange chromatography. However, this myeloperoxidase contained in addition one form with a lower and one form with a higher relative molecular mass, as indicated by gel-filtration chromatography. PMID- 3036099 TI - The role of ATP in the control of H+-galactoside symport in the yeast Kluyveromyces marxianus. AB - Transport of methyl beta-D-thiogalactoside and p-nitrophenyl beta-D-galactoside is shown to proceed through the H+-lactose symporter of Kluyveromyces marxianus. Uptake of these compounds is strongly reduced under anaerobic conditions or aerobically in the presence of antimycin. It is shown that antimycin treatment affects p-nitrophenyl beta-D-galactoside uptake in a similar way as it affects the cellular amount of ATP, suggesting regulation of p-nitrophenyl beta-D galactoside transport by ATP. Also, manipulation of cellular ATP by antimycin treatment followed by glucose incubation, or by aerobic incubation of cells with 2-deoxy-D-glucose, showed a similar dependence of galactoside uptake on the ATP level. Transport of the lipophilic cation tetraphenylphosphonium is affected by ATP variations in a similar way as galactoside influx. It is concluded that ATP regulates H+-galactoside symport by its influence on charge translocation. It is discussed that a membrane ATPase probably plays a central role in the control of the activity of H+-sugar symport. PMID- 3036100 TI - Effects of stimulation of muscarinic and of beta-catecholamine receptors on the intracellular distribution of protein kinase C in guinea pig exocrine glands. AB - Stimulation of exocrine cells via muscarinic receptors is associated with an activation of protein kinase C [Padel & Soling (1985) Eur. J. Biochem. 151, 1 10]. We show here that stimulation of isolated parotid gland lobules with 8 X 10( 6) M-carbamoylcholine leads to a translocation of protein kinase C from the cytosolic to the particulate compartment within 30 s (25% and 45% of total activity recovered in the particulate fraction of controls and stimulated samples respectively). The specific enzyme activity in the particulate fraction increased to 169% of the corresponding control value. After 10 min the changes started to reverse and, after 30 min, cytosolic protein kinase C was higher in stimulated than in unstimulated lobules. Isoproterenol (2 X 10(-5) M) stimulated the release of amylase more than did carbamoylcholine, but did not significantly affect intracellular distribution of protein kinase C during the observation time of 30 min. In isolated pancreatic lobules a significant carbamoylcholine-mediated translocation of protein kinase C into the particulate fraction could be observed after 5 and 20 min, but not after 1 min. After 5 min the specific enzyme activity in the particulate fraction had increased to 153% of the corresponding controls. The corresponding decrease (-38%) in the specific enzymic activity of cytosolic protein kinase C stayed constant up to 30 min. In isolated parotid gland lobules alpha-amylase secretion proceeded at a linear rate already during the first 1 min of stimulation, whereas in pancreatic lobules a measurable rate of alpha-amylase secretion did not occur before 5 min. These differences in time course paralleled the differences in the onset of translocation of protein kinase C. The results support a direct involvement of protein kinase C in carbamoylcholine-mediated but not in isoproterenol-mediated stimulation of exocytosis in exocrine cells. PMID- 3036101 TI - The regulation of phosphatidylcholine biosynthesis in rye (Secale cereale) roots. Stimulation of the nucleotide pathway by low temperature. AB - The incorporation of [14C]choline chloride and [14C]glycerol into segments taken from rye (Secale cereale L., cv. Rheidal) roots was greater in segments from roots grown at 5 degrees C than in segments taken from roots growing at 20 degrees C. The incorporation was measured at the temperature at which the root had been growing. Measurements in vitro of the enzymes of the nucleotide pathway showed activity of choline kinase (EC 2.7.1.32), choline-phosphate cytidylyltransferase (EC 2.7.7.15) and cholinephosphotransferase (EC 2.7.8.2) to be higher in homogenates from the cooler roots when assayed at 5 degrees C than the activities assayed at 20 degrees C in the 20 degrees C-root homogenates. Changes in vivo in the pool sizes of the CDP-base intermediates with temperature, relative differences in nucleotide-pathway-enzyme activities and a pulse-chase experiment with [14C]choline indicated that the rate-limiting step for phosphatidylcholine biosynthesis in this tissue, at both temperatures, was the reaction catalysed by cytidylyltransferase. PMID- 3036102 TI - Identification of signal sequence binding proteins integrated into the rough endoplasmic reticulum membrane. AB - An azidophenacyl derivative of a chemically synthesized consensus signal peptide has been prepared. The peptide, when photoactivated in the presence of rough or high-salt-stripped microsomes from pancreas, leads to inhibition of their activity in cotranslational processing of secretory pre-proteins translated from their mRNA in vitro. The peptide binds specifically with high affinity to components in the microsomal membranes from pancreas and liver, and photoreaction of a radioactive form of the azidophenacyl derivative leads to covalent linkage to yield two closely related radiolabelled proteins of Mr about 45,000. These proteins are integrated into the membrane, with large 30,000-Mr domains embedded into the phospholipid bilayer to which the signal peptide binds. A smaller, endopeptidase-sensitive, domain is exposed on the cytoplasmic surface of the microsomal vesicles. The specificity and selectivity of the binding of azidophenacyl-derivatized consensus signal peptide was demonstrated by concentration-dependent inhibition of photolabelling by the 'cold' synthetic consensus signal peptide and by a natural internal signal sequence cleaved and isolated from ovalbumin. The properties of the labelled 45,000-Mr protein-signal peptide complexes, i.e. mass, pI, ease of dissociation from the membrane by detergent or salts and immunological properties, distinguish them from other proteins, e.g. subunits of signal recognition particle, docking protein and signal peptidase, already known to be involved in targetting and processing of nascent secretory proteins at the rough endoplasmic reticulum membrane. Although the 45,000-Mr signal peptide binding protein displays properties similar to those of the signal peptidase, a component of the endoplasmic reticulum, the azido derivatized consensus signal peptide does not interact with it. It is proposed that the endoplasmic reticulum proteins with which the azidophenacyl-derivatized consensus signal peptide interacts to yield the 45,000-Mr adducts may act as receptors for signals in nascent secretory pre-proteins in transduction of changes in the endoplasmic reticulum which bring about translocation of secretory protein across the membrane. PMID- 3036103 TI - Involvement of phosphoinositide metabolism in potentiation by adrenaline of ADP induced aggregation of rabbit platelets. AB - Changes in phosphoinositide metabolism were examined in washed rabbit platelets stimulated with 0.5 microM-ADP, 50 microM-adrenaline, or ADP and adrenaline in combination. Adrenaline does not stimulate platelet aggregation when used alone, but does potentiate aggregation stimulated by ADP. In platelets prelabelled with [32P]Pi and [3H]glycerol, adrenaline was found to potentiate the ADP-induced changes in platelet phospholipids, causing larger increases in the amount and labelling of phosphatidylinositol 4-phosphate (PIP) and phosphatidic acid than was observed with ADP alone. The combination of ADP and adrenaline did not produce a greater decrease in phosphatidylinositol 4,5-bisphosphate (PIP2) than was produced by ADP alone. In platelets prelabelled with [3H]inositol, adrenaline potentiated the increases in labelling of inositol phosphate and inositol bisphosphate stimulated by ADP; no increase in inositol trisphosphate labelling was detected with ADP alone or with the combination of ADP and adrenaline. Phentolamine, an alpha-adrenergic-receptor antagonist, blocked potentiation by adrenaline of ADP-induced changes in phosphoinositide metabolism. Propranolol and sotalol, beta-adrenergic-receptor antagonists, augmented the potentiation; this is consistent with the concept that the effect of adrenaline is mediated by beta adrenergic receptors. The effect of adrenaline on phosphoinositide metabolism appears to be to potentiate the mechanisms by which ADP causes turnover of PIP and possibly degradation of PI, rather than the mechanism by which PIP2 is decreased. PMID- 3036106 TI - Molecular cloning of the human 2,3-bisphosphoglycerate mutase cDNA and revised amino acid sequence. AB - The human erythrocyte 2,3-bisphosphoglycerate mutase (BPGM) is a multifunctional enzyme which controls the metabolism of 2,3-diphosphoglycerate (DPG), the main allosteric effector of haemoglobin. Several cDNA banks were constructed from reticulocyte mRNA either by conventional cloning methods in plasmid pBR322 and screening with specific mixed oligonucleotide probes, or in the expression vector lambda gt 11. The largest cDNA isolated was 1673 bases, and encodes for a protein of 258 amino acids; it contains a large 3' untranslated region (785 bases). It is slightly smaller than the size of the intact mRNA estimated by Northern blot (1800 bases). Our sequence data indicate differences with the previously published amino acid sequence involving 21% of the residues. They were entirely confirmed by the amino acid composition of the tryptic peptides derived from purified BPGM. The revised amino acid sequence of the human BPGM is presented. PMID- 3036107 TI - Maturation dependence of the turnover of phosphatidylinositides in rabbit red blood cells. AB - "Pulse-chase" experiments with erythrocytes and reticulocytes prelabelled with 32P-Pi revealed a high turnover of the monoester phosphate groups of PIP2 in reticulocytes, which declines strongly during maturation [Maretzki et al., Biomed. Biochim. Acta 45, 1227-1236 (1986)]. The 3H-inositol uptake exhibits a strong maturational loss. A carrier-mediated uptake of inositol in reticulocytes is suggested. In reticulocytes PI, PIP and PIP2 incorporated 3H-inositol in the relative distribution of 86; 5 and 9%, respectively, but only PI to a minor extent in erythrocytes. A small release of 3H-labelled inositol phosphates was found in reticulocytes, which was not stimulated by extracellular calcium during incubation. Determination of the 32P/3H-ratio in double-labelled phosphatidylinositides in reticulocytes demonstrated a very slow turnover of the diester phosphate bonds compared with that of the inositol phosphate ester groups. PMID- 3036105 TI - Ca2+ uptake by corpus-luteum plasma membranes. Evidence for the presence of both a Ca2+-pumping ATPase and a Ca2+-dependent nucleoside triphosphatase. AB - Plasma-membrane vesicles from rat corpus luteum showed an ATP-dependent uptake of Ca2+. Ca2+ was accumulated with a K1/2 (concn. giving half-maximal activity) of 0.2 microM and was released by the bivalent-cation ionophore A23187. A Ca2+ dependent phosphorylated intermediate (Mr 100,000) was detected which showed a low decomposition rate, consistent with it being the phosphorylated intermediate of the transport ATPase responsible for Ca2+ uptake. The Ca2+ uptake and the phosphorylated intermediate (E approximately P) displayed several properties that were different from those of the high-affinity Ca2+-ATPase previously observed in these membranes. Both Ca2+ uptake and E approximately P discriminated against ribonucleoside triphosphates other than ATP, whereas the ATPase split all the ribonucleoside triphosphates equally. Both Ca2+ uptake and E approximately P were sensitive to three different Hg-containing inhibitors, whereas the ATPase was inhibited much less. Ca2+ uptake required added Mg2+ (Km = 2.2 mM), whereas the ATPase required no added Mg2+. The maximum rate of Ca2+ uptake was about 400-fold less than that of ATP splitting; under different conditions, the decomposition rate of E approximately P was 1,000 times too slow to account for the ATPase activity observed. All of these features suggested that Ca2+ uptake was due to an enzyme of low activity, whose ATPase activity was not detected in the presence of the higher-specific-activity Ca2+-dependent ATPase. PMID- 3036104 TI - Modulation by pterins of the phosphorylation and phenylalanine activation of phenylalanine 4-mono-oxygenase. AB - The interaction between phenylalanine 4-mono-oxygenase and analogues of the natural cofactor (6R)-tetrahydrobiopterin [(6R)-BH4] was studied. The rate of cyclic AMP-dependent phosphorylation of phenylalanine 4-mono-oxygenase was inhibited only by those pterins [(6R)-BH4, (6S)-BH4 and 7,8-dihydrobiopterin (BH2)] that were able to decrease the potency and efficiency of phenylalanine as an allosteric activator of the hydroxylase. Since BH2 lacks cofactor activity, this was not required to modulate either the phosphorylation or the phenylalanine activation of the hydroxylase. Half-maximal inhibition of the phosphorylation was observed at 1.9 microM-(6R)-BH4, 9 microM-(6S)-BH4 and 17 microM-BH2. Competition experiments indicated that all three pterins acted through binding to the cofactor site of the hydroxylase. Since the phosphorylation site and the cofactor binding site are known to reside, respectively, in the N- and C-terminal domains of the hydroxylase, the pterins were able to induce an interdomain conformational change. BH2, whose dihydroxypropyl group is not subject to epimerization, and (6S)-BH4 both inhibited the phosphorylation less efficiently than did the (6R) epimer of BH4. Pterins with different spatial arrangements of the dihydroxypropyl side chain thus appeared to elicit different conformations of the phosphorylation site. The hydroxylase reaction showed a higher apparent Km for (6S)-BH4 than for (6R)-BH4 both when the native and the phenylalanine-activated enzyme were tested. For the activated enzyme Vmax was 40% lower with the (6S)-epimer than the (6R) epimer, also when the more rapid enzyme inactivation occurring with the former cofactor was taken into account. PMID- 3036109 TI - Glycerated hemoglobin alpha 2 beta 2(82) (EF6) N-epsilon-glyceryllysine: a new post-translational modification occurring in erythrocyte bisphosphoglyceromutase deficiency. AB - A new minor Hb fraction initially designated Hbx, has been found in the hemolysate of a erythremic patient that we have previously described with a complete erythrocyte bisphosphoglyceromutase (BPGM) (E.C.2.7.5.4.) deficiency. Hbx (3.5% of the total) was detected by isoelectric focusing (IEF) and exhibited electrophoretic and chromatographic properties similar to that of several variants of the Hb central cavity. By density fractionation of red cells, it was demonstrated that Hbx was an aging hemoglobin as in the case of glycated Hb A1c. Functional studies revealed a low oxygen affinity and almost complete inhibition of the allosteric effect of the organic phosphate effectors. Structural studies demonstrated an absence of tryptic cleavage between the peptides beta T9 and beta T10 suggesting the presence of an adduct on Lys beta 82 or on a neighboring residue. FAB mass spectrometry, CID/MIKE spectra and a specific enzymatic assay with glyoxylate reductase, demonstrated that the 82 adduct was a glycerate moiety. It was concluded that Hbx was a glycerylated Hb: alpha 2 A beta 2(82) (EF6) N-epsilon-glyceryllysine, to our knowledge the first example of glycerylated protein. The mechanism of formation of glyceryl-Hb, which was found in the four studied subjects with a BPGM deficiency, remains to be determined. PMID- 3036108 TI - Alterations of red blood cell proteolysis in favism. AB - Damaged RBC drawn from favic patients during acute hemolysis showed marked alterations in their two major proteolytic systems. Cytosolic procalpain (i.e., the proenzyme species of Ca2+-activated neutral proteinase, or calpain) had considerably lower activity than in matched RBC from asymptomatic G6PD-deficient subjects. The total RBC activity of the three acid endopeptidases that are normally membrane-bound was not reduced in favism, but its subcellular distribution was mostly cytosolic, suggesting quantitative release from membranes. Changes in procalpain activity are the result of both autoxidation of divicine and of the intracellular elevation of Ca2+ that is found in favism. Changes in acid endopeptidase activity are the consequence of perturbed Ca2+ homeostasis. Overall, the picture shows a marked impairment of the RBC proteolytic machinery that in turn may worsen cellular damage. PMID- 3036110 TI - Metabolism of intact reticulocytes and mitochondria under lowered energy load induced by cycloheximide. AB - Cycloheximide was used to decrease the energy load by inhibitors of hemoglobin synthesis as the major ATP-consumer in intact rabbit reticulocytes. Inhibitor concentrations up to 2.5 X 10(-5) M led to decreased ATP-production at simultaneously elevated ATP/ADP ratios. Higher inhibitor levels resulted in further decreasing ATP-production but now in the reverse parallel to declining ATP/ADP ratios. Studies with intact reticulocyte mitochondria revealed almost no influence on respiration rates at the low range of concentrations but increasing inhibitions of state 3, state 4 and uncoupled respiration as well, with a 50% inhibition around 3 mM. Thus the dual action of the inhibitor is obviously due to the specific suppression of protein synthesis on the one hand and the unspecific inhibition of mitochondrial respiration on the other. Similar conclusions have been drawn by using ouabain to suppress the ATP-consumption by Na-K-ATPase. A preliminary load characteristic for rabbit reticulocytes with glucose as substrate is presented. PMID- 3036112 TI - Purine metabolism in normal and high-ITP human erythrocytes. Attempts to evaluate the ability to store the cells. AB - Storage of erythrocyte units from donors with ITP pyrophosphohydrolase deficiency have been studied and compared with units from normal donors. Verifying other investigations the incidence of this genetic disorder was found to be as high as about 3%. Hemolysis in the units was higher than in other units and there was a tendency to low total adenylate concentration. It is suggested that blood centers should organize a quality assurance program where one of the aims should be to detect genetic disorders that make the erythrocytes from the donors less suitable for long term liquid storage. PMID- 3036111 TI - A role of adenylate cyclase stimulation in energy metabolism of reticulocytes. AB - The effects of beta-adrenergic stimulation of adenylate cyclase on energy metabolism of rat reticulocytes were studied. A large initial rate of cAMP formation induced a rapid short-term stimulation of glycolysis which was followed by an exponential decline at maintained high cellular cAMP level. The decrease of glycolytic activity signifies a "metabolic desensitization" by a permanently increased cAMP concentration. Stimulation of PFK activity was found to be the main reason for the increased glycolytic rate which is due to the several fold increase of cAMP and in part to ATP-depletion. Total and coupled respiration, as well as the ATP/ADP ratio were not changed. The data indicate that stimulation of adenylate cyclase of reticulocytes selectively activates glycolysis, i.e. the cytosolic energy compartment with no detectable influence on oxydative phosphorylation. PMID- 3036113 TI - Reutilization pathway of purine nucleotides in human erythrocytes. AB - Human erythrocytes can operate the pyrophosphorolysis of IMP and utilize the phosphoribosyl moiety of this compound for the synthesis of AMP and of GMP from exogenous purine bases. Erythrocyte purine phosphoribosyltransferases seem to have a role in this interconversion pathway. PMID- 3036114 TI - Cyclosporin A inhibits kinase C-independent activation of the Na+/H+ exchanger by PDGF and vanadate. AB - Mitogen-induced activation of Na+/H+ exchange was studied in Swiss 3T3 fibroblasts. Phorbol myristic acetate (PMA) caused amiloride inhibitable cell alkalinization. PDGF and vanadate, but not bombesin or thrombin, caused additional alkalinization when given 10 min after a maximal dose of PMA. Down regulation of kinase C by 24 hr PMA exposure prevented the alkalinization response to bombesin and thrombin, but not to PDGF or vanadate. Cyclosporin A specifically blocked the additional alkalinization after PDGF or vanadate in cells acutely exposed to PMA and in kinase C down-regulated cells. Thus, there are at least two independent pathways which activate Na+/H+ exchange. PMA, bombesin, and thrombin act via kinase C. PDGF and vanadate cause additional stimulation of the Na+/H+ exchanger by a kinase C-independent pathway, inhibitable by cyclosporin A. PMID- 3036115 TI - Purification of 5'-nucleotidase from human placenta after release from plasma membranes by phosphatidylinositol-specific phospholipase C. AB - 5'-Nucleotidase was purified greater than 1000-fold from human placenta by treatment of plasma membranes with S. aureus phosphatidylinositol-specific phospholipase C and affinity chromatography on Con A Sepharose and AMP-Sepharose. The resulting enzyme had a specific activity of greater than 5000 mumol/hr/mg protein and a subunit molecular weight of 73,000. Goat antibodies against 5' nucleotidase inhibited enzyme activity and detected 5'-nucleotidase after Western blotting. These antibodies also recognized a soluble form of 5'-nucleotidase and residual membrane-bound 5'-nucleotidase which could not be released by phosphatidylinositol-specific phospholipase C treatment, suggesting that the three forms of the enzyme are structurally related. The soluble 5'-nucleotidase may be derived from the membrane-bound form by the action of an endogenous phospholipase C. The structural basis for the inability of some of the membrane bound 5'-nucleotidase to be released by phosphatidylinositol-specific phospholipase C is unknown. PMID- 3036116 TI - Size of the directing moiety at carbon 5 of cytosine and the activity of human DNA(cytosine-5) methyltransferase. AB - M13 DNAs in which carbon 5 of each deoxycytidine residue in one strand is replaced with a bulky group are very good substrates for human DNA (cytosine-5) methyltransferase. Rate enhancements of up to 35 fold are obtained depending on the size of the moiety at C-5. The enzyme appears optimally suited to sense a methyl group in one strand at this position. Alkaline density gradient analyses of the distribution of methyl groups applied to 5-BrdCyd or 5-IdCyd substituted DNA reveal that these groups serve to direct the enzyme to methylate the unsubstituted strand. PMID- 3036118 TI - Retention of enzymatic activity by N-terminal domain (1-78) T4-lysozyme: expression of synthetic DNA in Escherichia coli. AB - DNA of 235 b.p. coding for N-terminal domain (1-78) T4-lysozyme was synthesized and cloned by ligating twelve synthetic fragments with a linearized plasmid pUCE8 followed by transformation. On expression in E. coli strain JM103 cells, colonies containing the synthetic DNA were found to be lytic. On purification, clone ptly. 23-5 was found to contain polypeptide (M.W. 10,500), corresponding to N-terminal domain, its dimeric and aggregate form. It was identified by amino acid sequence analysis of the dimeric form. PMID- 3036117 TI - Reduced sensitivity to catecholamine in Werner's syndrome fibroblasts. AB - The beta-adrenergic receptor-coupled adenylate cyclase system has been investigated in normal and Werner's syndrome fibroblasts. The basal levels of cAMP in Werner and normal control cells were similar, whereas the isoproterenol induced increase in cAMP levels was far less for Werner cells than for control cells. In the broken cell preparations isoproterenol stimulated the adenylate cyclase of only control cells, not of Werner cells, although NaF or prostaglandin E1 stimulated the enzyme of both cells to the same extent. The beta-adrenergic receptor concentrations analyzed with hydrophilic radioligand were nearly equal in Werner and in control cells. A reduction of functional activity of the beta adrenergic receptor in Werner cells is thus suggested. PMID- 3036119 TI - The effect of EDTA-Fe(III) complexes with different chemical structure on the lipid peroxidation in brain microsomes. AB - Unlike EDTA-Fe(III) (1:1), the oxidized form of EDTA-Fe(II) complex enhanced lipid peroxidation in brain microsomes. Mossbauer spectroscopy and electron paramagnetic resonance analysis of oxidized EDTA-Fe(II), capable of inducing lipid peroxidation, showed the presence of EDTA-Fe(III) complex, which was different from the separately prepared (not oxidized) EDTA-Fe(III). Lipid peroxidation initiated by the oxidized EDTA-Fe(II) complex was dependent on the presence of NAD(P)H and functionally intact microsomes. No inhibitory effect was found by generally used free radical scavengers and catalase. Our results clearly indicate that the chemically different EDTA-Fe(III) complexes differ in their capability of initiating the NAD(P)H-dependent lipid peroxidation in brain microsomes. PMID- 3036120 TI - Similar physical and kinetic properties of rat brain synaptic membrane and cytosol phosphoinositide phospholipases C. AB - Phosphoinositide phospholipase C (PLC) was extracted from the synaptic membrane fraction of rat brain by 1% sodium deoxycholate. The molecular weight and sedimentation coefficient of the membrane PLC were about 160,000 and 6.7 as estimated by gel filtration and sucrose density gradient centrifugation, respectively. These values of the membrane PLC were identical with those of the cytosol PLC of the same tissue. Moreover, the membrane PLC showed the substrate specificity for phosphatidylinositol, phosphatidylinositol-4-monophosphate and phosphatidylinositol-4,5-bisphosphate and the sensitivity to Ca2+, sodium deoxycholate and N-ethylmaleimide similar to those of the cytosol PLC. These results indicate that the rat brain synaptic membrane PLC is indistinguishable from the cytosol PLC in physical and kinetic properties. PMID- 3036121 TI - Phosphocholinetransferase activity in plasma membrane: effect of diet. AB - The presence of phosphocholinetransferase, a component of the CDP-choline pathway for phosphatidylcholine biosynthesis is demonstrated in brain synaptic plasma membrane. Activity of phosphocholinetransferase is higher in weanling versus adult tissues, and responds to alterations in fat intake. The implications of phosphocholinetransferase activity in plasma membrane, and dietary manipulation of phosphatidylcholine biosynthesis via this pathway are discussed. PMID- 3036122 TI - Isolation of a cDNA clone for human cytochrome c1 from a lambda gt11 expression library. AB - Antiserum directed against a purified preparation of beef heart cytochrome bc1 complex has been used to screen a human liver cDNA expression library in lambda gt11. The inserts of two recombinants, which gave strong signals, were found to represent cytochrome c1 by epitope selection using nitrocellulose filters containing the expressed proteins. The amino acid sequence deduced from the nucleotide sequence of an insert DNA revealed a high degree of homology with the sequence of bovine cytochrome c1. PMID- 3036124 TI - Evidence that the lipolytic defect induced by estradiol-treatment in hamster adipocytes is related to an estrogen receptor-mediated defect in the adenylate cyclase catalytic subunit but not in Ns. AB - This study demonstrates that estradiol-treatment (10 micrograms per day x 5 days) does not impair the level of Ns, the adenylate cyclase stimulatory regulatory protein, in hamster fat cell membranes. In addition, this report shows that the defective cyclic AMP response induced in intact adipocytes by the estradiol treatment is either unaltered by the administration of alpha-bromocriptine or abolished by tamoxifen- or 4-hydroxytamoxifen-treatment. It can thus be concluded that the reduced lipolytic response found in hamster fat cells after estradiol treatment is related only to an estradiol-receptor-mediated defect in adenylate cyclase catalytic subunit activity which is independent from increased prolactin secretion. PMID- 3036123 TI - The adenosine receptor mediated accumulation of cyclic AMP in Jurkat cells is enhanced by a lectin and by phorbol esters. AB - The accumulation of cyclic AMP in Jurkat cells was stimulated by adenosine and adenosine analogues. The accumulation of cyclic AMP induced by these agents was competitively antagonized by the adenosine receptor antagonist 8-p-sulphophenyl theophylline (KD appr 1.9 microM). The lectin PHA, the diacylglycerol OAG as well as tumor promoting phorbol esters enhanced the accumulation of cyclic AMP induced by the adenosine analogue NECA. The results suggest that activation of CD2/CD3 receptors by lectins could potentiate the endogenous cyclic AMP stimulator adenosine via activation of protein kinase C. PMID- 3036125 TI - Cobalt-protoporphyrin causes prolonged inhibition of catechol estrogen synthesis by rat liver microsomes. AB - A single injection of cobalt-protoporphyrin (CoPP), which produces a marked and sustained decline in hepatic cytochrome P450 content, reduced the ability of male rat liver microsomes to form catechol estrogens to about 30% of control values within 1 day, as measured by the release of 3H2O from [2-3H]estradiol. Two days after treatment, the apparent Km of estrogen 2-hydroxylase for estradiol was increased, but other inhibitors of cytochrome P450 function (SKF-525A or piperonyl butoxide) failed to affect the enzyme. Inhibition by CoPP was also demonstrated by measuring the conversion of [4-14C]estradiol to its 2 hydroxylated derivative visualized by autoradiography after chromatographic separation. These findings point to yet another site in the multifaceted action of cobalt protoporphyrin. PMID- 3036126 TI - Yeast pyruvate carboxylase: gene isolation. AB - To improve our understanding of pyruvate carboxylase (PC)(EC 6.4.1.1) structure and the evolution of the biotin-dependent carboxylases we have isolated and sequenced a yeast (Saccharomyces cerevisiae) genomic DNA fragment encoding PC. The identity of the cloned gene was confirmed by comparing the encoded protein with the sequence of a 26 amino acid biotin-containing peptide isolated from yeast PC. The yeast PC sequence is homologous (43% amino acid homology) to the rat PC sequence, although the carboxyl-terminus was found to be 44 residues from the biotinyl-lysine whereas in all biotin carboxylases sequenced to date the biotin is 35 residues from the carboxyl-terminus. PMID- 3036127 TI - Conversion of estradiol-17 beta to reactive embryotoxic intermediates by cytochrome P-450-dependent bioactivating systems. AB - P-450-dependent enzyme systems added to media of cultured rat embryos markedly increased the embryotoxicity of estradiol-17 beta. Increases were markedly attenuated by omission of NADPH, omission of enzyme, substitution of female for male rat liver as enzyme source, d) replacement of N2 with CO or replacement of estradiol-17 beta with diethylstilbestrol. Embryotoxicity correlated well (r = 0.84) with catecholestrogen generating activities. Addition of a catechol methylating system failed to modify embryotoxicity even though large quantities of methoxyestrogens were formed. The results document that endogenous estrogen can be converted by P-450 to embryotoxic intermediates and suggest that reactive proximate metabolites are precatechols, perhaps epoxyenones. PMID- 3036128 TI - Cloned human interferons alpha: differential affinities for polyinosinic acid and relationship between molecular structure and species specificity. AB - The HuIFN-alpha A and HuIFN-alpha D interferons, produced by two independent recombinant bacterial clones, have different affinities for polyinosinic acid (poly I). The monomeric form HuIFN-alpha A (FMM), but not the HuIFN-alpha D, binds to poly (I)-agarose and is protected by poly (I) from thermal inactivation. Other subtypes of HuIFN-alpha A including the monomer SMM and oligomers have no affinity for this polynucleotide. In addition, these interferons show different target cell preferences in agreement with our previous suggestion (23) that the polynucleotide binding domain may be responsible for species specificity. Two significant observations are 1) the fractions of HuIFN-alpha D and HuIFN-alpha A unbound on poly (I)-agarose show higher antiviral inducing activity on heterologous (MDBK) than on homologous (WISH) cells, whereas they induce about the same activity of 2'5' oligoadenylate synthetase in these two cell lines. These fractions are also active on L929 cells. 2) The bound fraction of HuIFN alpha A induces almost the same antiviral and 2'5' oligoadenylate synthetase activities in MDBK and in WISH cells but neither activity in L929 cells. PMID- 3036129 TI - Evidence that the alpha and alpha (+)isoforms of the catalytic subunit of (Na+, K+)-ATPase reside in distinct ciliary epithelial cells of the mammalian eye. AB - Polyclonal antibodies against the canine kidney (Na+,K+)-ATPase were used to examine the localization and distribution of this protein in intact ciliary processes (CP) from bovine eyes by indirect immunofluorescence. The basolateral surface of non-pigmented (NPE) and pigmented (PE) ciliary epithelial cells was found to be stained specifically for the (Na+,K+)-ATPase. Immunoblot analysis of intact CP, separated PE and NPE cells by density gradients and cultured ciliary epithelial cells, revealed two forms of the catalytic subunit of the (Na+,K+) ATPase: the alpha and alpha (+). The alpha (+) form was enriched in NPE cells while alpha was in PE cells. PMID- 3036130 TI - Diferric transferrin reduction stimulates the Na+/H+ antiport of HeLa cells. AB - Proton release from HeLa cells is stimulated by external oxidants for the transplasmalemma electron transport enzymes. These oxidants, such as ferricyanide and diferric transferrin, also stimulate cell growth. We now present evidence that proton release associated with the reduction of ferricyanide and diferric transferrin is through the Na+/H+ antiport. The stoichiometry of H+/e- release with diferric transferrin is over 50 to 1, which is greater than expected for oxidation of a protonated transmembrane electron carrier. Diferric transferrin induced proton release depends on external sodium and is inhibited by amiloride. Proton release is also inhibited when diferric transferrin reduction is inhibited by apotransferrin. A tightly coupled association between the redox system and the antiport is shown by sodium dependence and amiloride inhibition of diferric transferrin reduction. The results indicate a new role for ferric transferrin in growth stimulation by activation of the sodium-proton antiport. PMID- 3036132 TI - The effects of DNA methylation by Hha I methylase on the cleavage reactions by Hae II, Aha II and Ban I endonucleases. AB - The DNA methylated by Hha I methylase was resistant against cleavage of Hae II or Aha II endonuclease indicating that the methyl group of the C5 position of the inmost cytosine nucleotide interferes with the interaction between the enzyme and the hexameric recognition sequence. Considering that Hae II or Aha II methylase has not been isolated yet, the result explained above is a useful information for protecting a double stranded DNA from being cleaved by Hae II or Aha II endonuclease. In contrast to Hae II or Aha II endonuclease, Ban I endonuclease which also has Hha I sequence as its tetrameric core was able to cleave the same DNA normally. This result suggests that the C5 position of the inmost pyrimidine nucleotide is not an important contact point between Ban I endonuclease and its hexameric recognition sequence. PMID- 3036133 TI - Tissue distribution of mRNAs encoding the alpha isoforms and beta subunit of rat Na+,K+-ATPase. AB - The tissue distribution of the multiple forms of rat Na+,K+-ATPase was examined at the molecular level with cDNA probes specific for the alpha, alpha (+), alpha III and beta subunit mRNAs. Northern and slot blot analyses demonstrate that these mRNAs are produced in a tissue-specific manner. RNAs encoding the alpha (+) isoform are detected in kidney, brain, heart, adipose, muscle, stomach and lung, whereas alpha III RNA is detected in brain, stomach and lung. Both alpha and beta mRNAs are present in all the tissues studied, although at very different levels. Examination of heart tissue in greater detail demonstrates that the levels of mRNA encoding the alpha subunit are greater in the atria than in the ventricles, while the converse is true for alpha (+). PMID- 3036131 TI - The basic glutathione S-transferases from human livers are products of separate genes. AB - We have characterized a second cDNA sequence, pGTH2, for the human liver glutathione S-transferases Ha subunits. It is 95% homologous base-for-base to the Ha subunit 1 cDNA, pGTH1, except for its longer 3' noncoding sequences. Our results indicate that the multiple basic human liver glutathione S-transferases are products of separate genes. The proposal [Kamisaka, K., Habig, W. H., Ketley, J. N., Arias, I. M., and Jakoby, W. B. (1975) Eur. J. Biochem. 60, 153-161] that deamidation may be a physiologically important process for generating glutathione S-transferases isozyme multiplicity can be all but ruled out. PMID- 3036135 TI - The metabolism of inositol 4-monophosphate in rat mammalian tissues. AB - Rat brain soluble fraction contains an enzymatic activity that dephosphorylates inositol 1,4-bisphosphate (Ins(1,4)P2). We have used anion exchange h.p.l.c. in order to identify the inositol monophosphate product of Ins(1,4)P2 hydrolysis (i.e. Ins(1)P1, Ins(4)P1 or both). When [3H]Ins(1,4)P2 was used as substrate, we obtained an inositol monophosphate isomer that was separated from the co-injected standard [3H]Ins(1)P1. This suggested an Ins(1,4)P21-phosphatase pathway leading to the production of the inositol 4-monophosphate isomer. The dephosphorylation of [32P]Ins(4)P1 was measured in rat brain, liver and heart soluble fraction and was Li+-sensitive. Chromatography of the soluble fraction of a rat brain homogenate on DEAE-cellulose resolved a monophosphate phosphatase activity that hydrolyzed both [3H]Ins(1)P1 and [4-32P]Ins(4)P1 isomers. PMID- 3036134 TI - Specific binding of atrial natriuretic factor to renal glomeruli in Doca- and Doca-salt-treated rats correlation with atrial and plasma levels. AB - Since volume expansion and high blood pressure (BP) are known stimuli of atrial natriuretic factor (ANF) release, and since this peptide may be involved in mineralocorticoid escape, we investigated the effects of chronic deoxycorticosterone (DOCA) and DOCA-NaC1 treatment on renal glomerular ANF receptor density and affinity in relation to atrial and plasma ANF levels. An increase in plasma immunoreactive ANF (IR-ANF) was observed both after two and four weeks of treatment. IR-ANF concentrations were elevated in the left atrium only in four-week DOCA treated rats. Administration of the mineralocorticoid alone resulted in a decreased density of glomerular ANF receptors in both time periods investigated. DOCA-NaC1-treated animals presented an increased receptor density during the pre-hypertensive stage (2 weeks) and a reduced density in the later hypertensive period (4 weeks). Receptor affinity in both groups was identical to that in the controls after 2 weeks and was augmented after 4 weeks of treatment. Our data suggest that the down-regulation of renal glomerular ANF receptors during chronic DOCA-NaC1 administration may play a role in the maintenance of high BP in this model of volume-expanded hypertension. PMID- 3036136 TI - Effects of experimental acute pancreatitis in dogs on metabolism of lung surfactant phosphatidylcholine. AB - Acute haemorrhagic pancreatitis was produced in the dogs by transduodenal injection of autologous bile into the main pancreatic duct. There was no significant change in the activity of three regulatory enzymes of phosphatidylcholine biosynthesis (glycerophosphate acyltransferase, cytidyltransferase and cholinephosphotransferase) in lung; however, there was a 42% decrease in the amount of dipalmitoyl phosphatidylcholine (surfactant) in lung lavage due to acute pancreatitis. The decrease in lavage phospholipid content was associated with 5-fold increase in phospholipase A2 activity of lung lavage, and massive accumulation of osmiophilic spheroid structures in the alveolar space. PMID- 3036137 TI - Direct electron transfer reactions of cytochrome c553 from Desulfovibrio vulgaris Hildenborough at indium oxide electrodes. AB - The direct, heterogeneous, electron transfer reactions of cytochrome c553 from Desulfovibrio vulgaris Hildenborough have been studied at indium oxide optically transparent electrodes. These reactions have been studied using cyclic voltammetry and derivative cyclic voltabsorptometry and the kinetics of heterogeneous electron transfer is quasi-reversible. The thermodynamics and kinetics of electron transfer by this molecule can be studied at this electrode surface without the need for surface modification or the addition of surface promoters or mediators. PMID- 3036138 TI - Binding and action of glucagon in isolated adipocytes from cortisol-treated rats. AB - Evidence for pre-receptor, receptor and post-receptor glucagon defects was investigated in adipocytes from cortisol-treated rats. A decrease in glucagon binding due to a decreased number of receptors was observed. No changes in receptor affinity were detected. Both, the lipolytic response of glucagon and the ability of glucagon to increase basal and theophylline-stimulated cAMP accumulation remained unaltered. Moreover, a hyperglucagonemia accompanied by an increase in glucagon degradation in the serum of cortisol-treated rats was observed. Such alterations could represent a new mechanism by which glucocorticoids exert their biological actions. PMID- 3036139 TI - In vitro synthesis of 32P-labelled phosphatidylinositol 4,5-bisphosphate and its hydrolysis by smooth muscle membrane-bound phospholipase C. AB - For studies of phospholipase C (PLC) activity in cell-free systems, 32P-labelled phosphatidylinositol 4,5-bisphosphate (PIP2) was prepared enzymatically by phosphorylating phosphatidylinositol 4-phosphate (PIP) in the presence of [gamma 32P]ATP using a PIP kinase partially purified from bovine retinae. PLC activity was determined by incubating membranes of DDT1 MF-2 cells with 32P-PIP2 and measuring remaining non-hydrolyzed substrate as well as accumulation of the hydrolysis product, inositol trisphosphate (IP3). Guanine nucleotides stimulated PIP2 hydrolysis and IP3 release. Additional increase in IP3 accumulation was observed with adrenaline plus guanine nucleotides. PMID- 3036140 TI - Atrial natriuretic peptide, ANP(99-126), receptors in rat thymocytes and spleen cells. AB - A single class of saturable, specific binding sites for the circulating form of atrial natriuretic peptides, ANP(99-126), was identified in rat thymus and spleen and in isolated thymocytes and spleen cells using quantitative autoradiographic techniques. In the thymus, the relative potency of ANP analogs to inhibit [125I] ANP(99-126) binding was ANP(99-126) = ANP(103-126) greater than ANP(111-126) greater than ANP(103-125). ANP(103-123) could not displace [125I]ANP(99-126) binding. Addition of ANP(99-126) stimulated the formation of cyclic GMP in isolated thymocytes and spleen cells in a dose-dependent manner. Our results indicate that immune cells have specific ANP receptors which could be coupled to guanylate cyclase activation and may play a role in the regulation of the immune response. PMID- 3036142 TI - Water induced dismutation of superoxide anion generates singlet molecular oxygen. AB - Direct spectroscopic measurement of 1268 nm singlet oxygen emission from KO2 suspensions at room temperature in three non-protonic solvents--CCl4, Cl2FCCClF2, and C6F14 by the action of water is reported. The results clearly show that the singlet oxygen generation is due to a water induced reaction, and suggest that one role of the enzyme superoxide dismutase may be the protection of biological structures, for example, lipid membranes, from degradation by singlet oxygen. PMID- 3036141 TI - Chemotactic peptide, calcium and guanine nucleotide regulation of phospholipase C activity in membranes from DMSO-differentiated HL60 cells. AB - Membranes prepared from DMSO-differentiated HL60 cells labeled with [3H]inositol hydrolyze polyphosphoinositides in a Ca2+-dependent manner, generating inositol 1,4-bisphosphate (IP2) and inositol 1,4,5-trisphosphate (IP3). Incubation of membranes with GTP or GTP gamma S reduces the concentration of Ca2+ required for activation. This nucleotide effect is potentiated by formyl-Met-Leu-Phe (FMLP). Pertussis toxin inhibits FMLP-induced augmentation, but not the induction of IP2/IP3 formation by GTP or GTP gamma S. These results suggest that differentiated HL60 cells contain a membrane-associated phospholipase C that degrades polyphosphoinositides and that activation of this enzyme is mediated by at least two guanine nucleotide binding proteins, one of which is linked to FMLP receptors and is pertussis toxin sensitive. PMID- 3036143 TI - Identification of the product of dnaB gene in Bacillus subtilis. AB - In order to detect the product of dnaB gene in B. subtilis, a gene which is involved in the initiation of DNA replication and the formation of the DNA membrane complex, we synthesized an origopeptide of 15 amino acids which corresponds to a region near the carboxyl-terminal of the gene product, and raised antibody against the synthetic peptide. We have also employed a filter binding assay to measure the predicted DNA binding activity of the product of the dnaB gene, using the plasmid pUB110. The binding activity was detected after fractionation of cell lysates of B. subtilis on sucrose-density gradients. When the active fraction was prepared from a mutant which was temperature-sensitive for the dnaB gene, the DNA binding activity in the fraction showed significant thermolability. Furthermore, the binding activity was inhibited by the purified antibody raised against the synthetic peptide. These results suggest that the product of the dnaB gene does indeed have DNA binding activity, and that the filter binding assay and the antibody can be used for the detection and characterization of the gene product. PMID- 3036144 TI - Spectral properties of nitric oxide complex of cytochrome c' from Rhodopseudomonas capsulata B100. AB - The spectral properties for NO complexes of ferric and ferrous cytochrome c' from photosynthetic bacterium Rhodopseudomonas capsulata B100 are reported. The electronic absorption, MCD, and EPR spectra have been compared with those of the NO complexes of the other cytochromes c' and horse heart cytochrome c. The NO ferrous cytochrome c' would be a mixture of NO complexes with six- and five coordinate nitrosylheme, suggesting that the heme-iron to histidine bond in the ferrous cytochrome c' is more stable than that from chemoheterotrophic bacteria. The reaction product of ferric cytochrome c' with NO exhibited the spectra similar to NO-ferric derivatives of the other hemoproteins, which indicates the formation of NO-ferric cytochrome c'. PMID- 3036145 TI - Isolation of a cDNA clone for the dihydrolipoamide acetyltransferase component of the human liver pyruvate dehydrogenase complex. AB - Dihydrolipoamide acetyltransferase (E2) forms the structural core of pyruvate dehydrogenase complex. A cDNA clone (lambda E2-1) for mammalian E2 was identified from a human liver lambda gt11 library using anti-E2 serum. Affinity-selected antibodies using the fusion protein from lambda E2-1 immuno-reacted specifically with E2 of purified pyruvate dehydrogenase complex on immuno-blot analysis. The cDNA insert was approximately 2.3 kb in length with an internal EcoR1 site generating 1.4 and 0.9 kb fragments. A synthetic 17-mer oligodeoxynucleotide mixture based on the amino acid sequence surrounding the lipoic acid-containing lysine residue in bovine kidney E2 hybridized with the 2.3 kb cDNA insert and the 1.4 kb fragment. PMID- 3036146 TI - Signalling for increased cytoskeletal actin in neutrophils. AB - The addition of fMet-Leu-Phe, platelet-activating factor, leukotriene B4 or sodium propionate to rabbit neutrophils causes an increase in the amount of actin associated with the cytoskeletal actin. The increase is rapid, transient and inhibitable by pertussis toxin. On the other hand, the addition of phorbol 12 myristate 13-acetate or NH4Cl causes a pertussis toxin-insensitive increase in cytoskeletal actin. The effects of the phorbol ester and fMet-Leu-Phe are additive, and in the presence of the phorbol ester, the fMet-Leu-Phe induced effect declines to the level produced by the phorbol ester. These results suggest that: one of the signalling pathways for actin polymerization involves a guanine nucleotide binding protein; actin polymerization mediated through this pathway is rapid, transient and inhibitable by pertussis toxin, and a second signalling pathway is independent of this guanine-nucleotide binding protein; actin polymerization, mediated by this second pathway, is somewhat slower, sustained and insensitive to pertussis toxin. These results are discussed in terms of a model which includes gelsolin, profilin and the pertussis toxin-sensitive guanine nucleotide binding protein. PMID- 3036147 TI - Sequence homologies between p36, the substrate of pp60src tyrosine kinase and a 67 kDa protein isolated from bovine aorta. AB - A 67 kDa actin-binding protein was isolated from bovine aorta. Partial amino acid sequence determination of two large thermolysin peptides were used to compare 67 kDa bovine aorta protein and p36 the substrate of pp60src tyrosine kinase. Sequence analysis shows that 67 kDa bovine aorta protein shares common domains with p36 and possesses the consensus aminoacid sequences of mammalian Ca2+ dependent membrane-binding protein and p36/gelsolin. PMID- 3036148 TI - Phospholipid dependence of rat brain microsomal glucose-6-phosphate phosphohydrolase. AB - Partial lipid removal of rat brain microsomes by acetone-butanol extraction resulted in 32% loss of activity of glucose-6-phosphate phosphohydrolase (G-6 Pase) and an increase in Km and energy of activation (Ea) of the enzyme while the Vmax was lowered. The activity was restored by supplementation of microsomal total phospholipid (PL) and phosphatidylcholine (PC) in sonicated dispersions but not with neutral lipids, phosphatidyl ethanolamine, sphingomyelin, phosphatidylglycerol and cholesterol. In both intact and delipidated membranes, the activity was decreased by sodium deoxycholate and enhanced by dimethylsulfoxide. Egg yolk PC and asolectin influenced the activity to the extent of that produced by microsomal PC. PC increased the Km of the enzymatic reaction in intact microsomes but decreased the same in disrupted membrane while the Vmax was not affected in both the membranes. Addition of PC into the assay system lowered Ea of the reaction in both the membrane systems. However, there was no break observed in the Arrhenius plot. Ability of liver nonspecific lipid transfer proteins to introduce alien PL into brain microsomes was used to study lipid dependence of G-6-Pase and investigation of membrane-enzyme interrelationship. Protein catalyzed transfer of egg PC from a donor PC cholesterol unilamellar liposomes resulted in substantial increase in microsomal membrane PC and total PL and a net reduction in the enzyme activity was observed in intact and delipidated membranes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3036149 TI - Preparation of spin-labelled sulfatides for EPR studies on model membranes. AB - Using the N-hydroxysuccinimide ester of the fatty acids, galactosylceramide I3 sulfate containing a 5-or 16-doxyl-stearoyl residue was prepared in good yield by acylation of the galactosylsphingosine I3-sulfate (lysosulfatide) obtained from the saponification of the bovine brain sulfatide. The EPR behavior of the two semisynthetic sulfatides was analyzed in natural sulfatide micelles and in multilamellar vesicles of egg phosphatidylcholine. The evaluated parameters demonstrate that these spin-labelled sulfatides can be used for the study of sulfatide behavior in lipid structures. PMID- 3036150 TI - Effects of cell isolation procedures and radioligand selection on the characterization of human leukocyte beta-adrenergic receptors. AB - Radioligand binding techniques are commonly used in the characterization of beta adrenergic receptors on human peripheral leukocytes. Accurate interpretation of receptor binding parameters necessitates appropriate radioligand selection. In addition, cell isolation techniques should have minimal effect on the binding parameters of receptors. Our observation of curvilinear Scatchard plots with (-) [125I]iodocyanopindolol (ICYP) resulted in a re-evaluation of this radioligand and the influence of cell isolation techniques on leukocyte beta-adrenergic receptor binding parameters. Membranes from mononuclear (MN) and polymorphonuclear (PMN) cells isolated by a standard procedure (Ficoll-Hypaque) resulted in biphasic Scatchard plots with ICYP in three of four subjects. In contrast, linear Scatchard plots were observed for ICYP binding to membranes from MN and PMN cells isolated from the same four subjects with an alternative procedure utilizing plasma Percoll. Competition and saturation binding assays with ICYP identified a high degree of nonspecific binding. Decreased stereoselectivity with (-)- and (+)-propranolol was observed with membranes from Ficoll-Hypaque cells as compared to plasma Percoll cells. Kinetic analysis with ICYP demonstrated apparent irreversible binding whether displacement was initiated with a beta-adrenergic receptor antagonist or agonist. These problems with ICYP prompted evaluation of an alternative radioligand, (-) [125I]iodopindolol (IPIN); this radioligand demonstrated rapid and completely reversible binding, improved stereoselectivity, and low nonspecific binding. Using IPIN, Scatchard plots from three additional subjects were linear for both cell isolation procedures. Based on these observations, the preferred method of human leukocyte beta-adrenergic receptor analysis incorporates the plasma Percoll cell isolation technique and the radioligand IPIN. PMID- 3036151 TI - Inhibition of metabolic response of polymorphonuclear leukocyte by biscoclaurine alkaloids. AB - Effects of biscoclaurine alkaloids on the various stimulus-responses of PMN, especially on the O2-. generation of PMN, were investigated. Results obtained were: cepharanthine inhibited various metabolic responses of PMN, its biological action probably being due to its membrane modifying action. Inhibition of O2-. generation by cepharanthine was stronger than any other inhibition of metabolic responses of PMN. The inhibitory effect of various biscoclaurine alkaloids on the O2-. generation of PMN was the descending order of tri-, di- and mono-ether type; the coclaurine type showed only a weak effect. PMID- 3036152 TI - Stimulation of prostacyclin production in blood vessels by the antithrombotic drug suloctidil. AB - Suloctidil is a calcium antagonist with vascular relaxing activity and an antithrombotic agent: its antiplatelet action has been demonstrated in vivo, but is difficult to reproduce in vitro and the mechanism of this effect remains unknown. We have observed that suloctidil (10 microM) stimulated the release of prostacyclin (PGI2) from the rabbit aorta, the dog vena cava and the dog portal vein, in vitro. This effect could be explained by an increased mobilization of free arachidonic acid. Neither the inactive congener CP894S, nor the two calcium channel antagonists, verapamil and flunarizine, reproduced the stimulatory effect of suloctidil. Suloctidil acted selectively on the vascular endothelium: it stimulated the release of PGI2 from bovine aortic and human umbilical vein endothelial cells, but neither from the de-endothelialized rabbit aorta nor from the bovine aortic media. The stimulatory effect of suloctidil on the release of the platelet inhibitor PGI2 from the vascular endothelium might contribute to the known antiplatelet and antithrombotic activity of this drug. PMID- 3036153 TI - Molecular probes for extracellular adenosine receptors. AB - Derivatives of adenosine receptor agonists (N6-phenyladenosines) and antagonists (1,3-dialkyl-8-phenylxanthines) bearing functionalized chains suitable for attachment to other molecules have been reported [Jacobson et al., J. med. Chem. 28, 1334 and 1341 (1985)]. The "functionalized congener" approach has been extended to the synthesis of spectroscopic and other probes for adenosine receptors that retain high affinity (Ki approximately 10(-9)-10(-8) M) in A1 receptor binding. The probes have been synthesized from an antagonist xanthine amine congener (XAC) and an adenosine amine congener (ADAC). [3H]ADAC has been synthesized and found to bind highly specifically to A1-adenosine receptors of rat and calf cerebral cortical membranes with KD values of 1.4 and 0.34 nM respectively. The higher affinity in the bovine brain, seen also with many of the probes derived from ADAC and XAC, is associated with phenyl substituents. The spectroscopic probes contain a reporter group attached at a distal site of the functionalized chain. These bifunctional ligands may contain a spin label (e.g. the nitroxyl radical TEMPO) for electron spin resonance spectroscopy, or a fluorescent dye, including fluorescein and 4-nitrobenz-2-oxa-1,3-diazole (NBD), or labels for 19F nuclear magnetic resonance spectroscopy. Potential applications of the spectroscopic probes in characterization of adenosine receptors are discussed. PMID- 3036154 TI - Presence of endogenous digitalis-like activity in mammalian heart not due to fatty acids. PMID- 3036155 TI - Arachidonic acid monooxygenase and lipoxygenase activities in polymorphonuclear leukocytes. PMID- 3036156 TI - Beta 1-adrenergic selectivity of the new cardiotonic agent denopamine in its stimulating effects on adenylate cyclase. AB - Effects of the new selectively beta 1-adrenergic cardiotonic drug denopamine (TA 064), (-)-(R)-1-(p-hydroxyphenyl)-2-[(3,4-dimethoxyphenethyl)amino]ethanol, on the adenylate cyclase-adenosine-3',5'-monophosphate (c-AMP) system of various tissues and cells in rats and guinea pigs were investigated in comparison with those of isoproterenol. Denopamine at concentrations above 10(-6) M stimulated lipolysis in vitro, and, above 10(-5) M, elevated the c-AMP level in isolated rat fat cells. The c-AMP level of guinea-pig heart ventricular muscle was also elevated when the heart was perfused with 3 X 10(-6) M denopamine or when slices of ventricular muscle were incubated with 10(-6) M denopamine. These changes were abolished in the presence of beta-adrenergic antagonists. Incubation with denopamine did not cause substantial elevation of c-AMP levels in rat reticulocytes and diaphragm. Denopamine activated adenylate cyclase of the rat cell membranes in a concentration-dependent manner. Although dose dependence was less apparent, denopamine also activated adenylate cyclase of the membrane fraction from guinea pig cardiac muscle, but it hardly activated the same enzyme from rat reticulocytes. Isoproterenol, on the other hand, showed marked concentration-dependent activation of adenylate cyclase in all these preparations. Denopamine did not inhibit c-AMP phosphodiesterase of both particulate and supernatant fractions of guinea-pig cardiac muscle. The stimulation of lipolysis by denopamine was observed even when elevation of the c AMP level was not detected, while the stimulation of lipolysis by isoproterenol was always accompanied with an elevation of c-AMP. When guinea-pig hearts were perfused with 3 X 10(-6) M denopamine or 10(-7) M isoproterenol, their cardiotonic effects were of the same magnitude whereas the degree of c-AMP elevation in the ventricular tissue by denopamine was significantly less than that by isoproterenol. It was concluded that stimulation of the adenylate cyclase c-AMP system by denopamine was restricted to the tissues whose receptors were predominantly of the beta 1-type, and that the elevation of c-AMP levels in these tissues by denopamine was less marked than by isoproterenol, suggesting that the stimulation of lipolysis and heart by denopamine may be mediated by a special pool of c-AMP or some other unknown factor(s). PMID- 3036157 TI - Comparative effects of calmodulin inhibitors on calmodulin's hydrophobic sites and on the activation of cyclic nucleotide phosphodiesterase by calmodulin. AB - Experiments were designed to investigate the effect of inhibitors on calmodulin's hydrophobic sites and their consequences on the activation of a target enzyme, cyclic nucleotide phosphodiesterase. Two fluorescent probes, 2-(p-toluidinyl) naphthalene-6-sulfonate (TNS) and 9-anthroylcholine (9AC) were used to study the interactions with calmodulin of inhibitors devoid of direct effect on the probes. Contrary to W-7, nicergoline, nicardipine and quercetin, which decreased the fluorescence of the two probes bound to calmodulin, bepridil only decreased 9AC fluorescence but increased the fluorescence intensity at the wavelength of the emission maximum of TNS. In spite of this difference, bepridil as well as W-7 and nicergoline competitively inhibited calmodulin activation of phosphodiesterase. In addition, nicergoline also inhibited phosphodiesterase activity competitively to cyclic GMP. These results show differences in the interactions of inhibitors with calmodulin; these differences are not detected in functional studies of the effect of inhibitors on phosphodiesterase activation. PMID- 3036158 TI - Elimination of isoproterenol-induced proline-rich protein biosynthesis in rat salivary glands after adult thyroidectomy. AB - Surgical thyroidectomy of adult rats resulted in a gradual decrease in beta adrenergic receptor density on the cell surface of parotid and submandibular glands. The decrease in beta-adrenergic receptors was 29 and 50% by 2 and 4 weeks, respectively, for the parotid gland. In the submandibular gland, a decrease of 50% of the total beta-receptor density was evident after 2 weeks. After 4 weeks no further decrease in beta-adrenergic receptor was observed. Subsequent challenge of the salivary glands with chronic treatment of isoproterenol (beta-adrenergic receptor agonist) failed to induce proline-rich protein and glycoprotein biosynthesis in the submandibular gland 2 weeks after thyroidectomy, whereas the parotid gland showed induced proline-rich protein but not the glycoprotein synthesis. By 4 weeks the parotid gland did not show the induced synthesis of the glycoprotein or proline-rich proteins. The inability to induce protein synthesis was reflected by decreased cAMP accumulation in both glands after injection with isoproterenol. Partial reversal of these effects on protein synthesis and cAMP accumulation was obtained by triiodothyronine treatment of thyroidectomized rats. PMID- 3036159 TI - Effects of ethanol on ouabain inhibition of mouse brain (Na+,K+)ATPase activity. AB - Plots of ouabain inhibition of mouse cerebral cortical (Na+,K+)ATPase activity fitted a two-site model significantly better than a one-site model, consistent with the presence of two forms of the enzyme with different affinities for ouabain. The fraction of enzyme activity with high affinity for ouabain (HAO: Ki = 500 nM), suggested to be localized neuronally, constituted the major portion (60-70%) of activity. Ouabain inhibition of both components of enzyme activity was reduced as KCl concentrations were increased. In vitro, only high concentrations of ethanol affected (Na+,K+)ATPase activity and ouabain inhibition of activity. Ethanol (500 mM) selectively reduced the activity, and increased the sensitivity to ouabain inhibition, of the HAO component, with no significant effect on the low-affinity (LAO) component. On the other hand, following chronic treatment of mice with ethanol in vivo, in a paradigm that produced tolerance and physical dependence, the sensitivity to ouabain of the HAO form of the enzyme was selectively increased. The relative proportions, and the activities of the HAO and LAO components, were not altered. The effects of ethanol, added in vitro, on the HAO component were decreased in ethanol-tolerant animals. The selective effect of chronic ethanol ingestion on (Na+,K+)ATPase activity indicates the specificity of action of ethanol in the CNS. PMID- 3036160 TI - Monoclonal antibodies specific for 1-4 benzodiazepines. PMID- 3036161 TI - Mechanism of inverse regulation of alpha 1- and beta-adrenergic receptors. PMID- 3036163 TI - Inhibition of neutrophil response by mepacrine. AB - Clinical and experimental evidence supports neutrophil involvement in the pulmonary complications of adult respiratory distress syndrome. Preliminary evidence indicates that mepacrine salvages pulmonary function in experimental models of adult respiratory distress syndrome, possibly by inhibiting neutrophil activation [E. M. Canham et al., Am. Rev. resp. Dis. 127, 594 (1983)]. This study examines the effect of mepacrine on neutrophil responses involved in pulmonary dysfunction associated with adult respiratory distress syndrome. A comparison is made between the ability of mepacrine to inhibit a specific neutrophil response utilizing different stimuli and the ability to inhibit different neutrophil responses to a single stimulus. Neutrophils were activated by a soluble stimulus, phorbol myristic acetate, and a particulate stimulus, heat-inactivated opsonized group B streptococcus. Mepacrine inhibited superoxide production in response to both phorbol myristic acetate (IC50 = 5.3 +/- 1.2 microM) or opsonized group B streptococcus (16.1 +/- 1.7 microM). Chemotaxis in response to n formylmethionylleucylphenylalanine was also inhibited (41.1 +/- 2.2 microM). Finally, aggregation stimulated by either streptococcus or phorbol myristic acetate was inhibited by mepacrine (73.0 +/- 9.8 microM and 77.0 +/- 19.2 microM respectively). A comparison of the IC50 values demonstrates that the inhibitory effect of mepacrine is response dependent and stimulus independent. The results of this study are consistent with the proposal that mepacrine protects against the pulmonary complications associated with adult respiratory distress syndrome by its action as an inhibitor of neutrophil function. PMID- 3036162 TI - Effect of ketoconazole on cholesterol synthesis and on HMG-CoA reductase and LDL receptor activities in Hep G2 cells. AB - Ketoconazole, an imidazole derivative, is a member of a class of metabolic inhibitors acting specifically at cytochrome-P450 mediated reactions. We studied the effects of this compound on cholesterol synthesis, and on HMG-CoA reductase and LDL receptor activities, in cultures of human hepatoma cell line Hep G2. Ketoconazole, added in concentrations of 2-100 microM, inhibited cholesterol synthesis, and caused accumulation of lanosterol and dihydrolanosterol. Total mass formation of sterols was depressed. After 20 hr preincubation of the cells with the drug in these concentrations, activity of HMG-CoA reductase was markedly decreased, while the receptor-mediated binding, uptake and degradation of human LDL were increased. This increase is at least partly due to a higher affinity of LDL for its receptor. Ketoconazole prevented the fall in LDL-receptor activity caused by preincubation with LDL, whereas it did not affect the suppression caused by preincubation with exogenous mevalonate. These findings are discussed with respect to the involvement of endogenous sterol and non-sterol effectors of reductase and receptor activities. PMID- 3036164 TI - Beta-adrenergic responsiveness and cardiac autonomic receptors after implantation of the MtTW15 pituitary adenoma in the rat. AB - The effects of chronic MtTW15 pituitary adenoma implantation on beta-adrenergic responsiveness, cardiac beta-adrenoreceptors, and muscarinic receptors were studied in the rat. Five weeks after s.c. administration of tissue fragments of the MtTW15 adenoma, there was a 51 and 20% increase in the heart weight and body weight, respectively, and a 49-fold increase in the serum prolactin level as compared to the controls. At this time there was also an attenuation in the adenoma-bearing group of the ability of isoproterenol to produce a dipsogenic response and to increase the heart rate. In contrast, isoproterenol stimulated cardiac ornithine decarboxylase (ODC) activity 4.2-fold in both the control and adenoma-bearing groups. There was no change between the two groups in the cardiac ventricular beta-adrenoreceptor or muscarinic receptor concentration as measured by specific (-)-[125I]iodocyanopindolol (CYP) and (-)-[3H]quinuclidinyl benzilate (QNB) respectively. In addition, the concentrations of isoproterenol and carbachol required to inhibit by 50% (IC50) [125I]CYP and [3H]QNB binding, respectively, in the absence of 5'-guanylylimidodiphosphate (Gpp(NH)p) were not different between the two groups. In the presence of Gpp(NH)p, the isoproterenol IC50 value was not different between the two groups, whereas the carbachol IC50 value was increased slightly in the adenoma-bearing group. The data indicate that chronic MtTW15 adenoma implantation attenuated beta-mediated dipsogenic and heart rate responses but had little or no effect on cardiac ODC activity or cardiac autonomic receptor concentrations and agonist binding properties. PMID- 3036165 TI - Down-regulation of surface beta-adrenoceptors on intact human mononuclear leukocytes. Time-course and isoproterenol concentration dependence. AB - Incubation of human mononuclear leukocytes (MNL) in vitro with isoproterenol resulted in a rapid loss of surface beta-adrenoceptors, determined by radioligand binding at 4 degrees. Isoproterenol concentrations in the range of 10 nM to 100 microM resulted in significant down-regulation of beta-adrenoceptors. At a concentration of 1 microM isoproterenol, the time-dependent loss of surface beta adrenoceptors closely paralleled the loss in isoproterenol-stimulated adenylate cyclase activity. If receptor number in intact cells was determined at 32 degrees, hardly any loss in receptor number was observed, due to reversal of down regulation during the incubation period. When beta-adrenoceptor number in broken cell preparations was determined by [125I]cyanopindolol binding at 37 degrees no significant loss was observed, even after 2 hr of isoproterenol treatment, while [3H]CGP-12177 binding resulted in a similar reduction in binding sites as in intact cells. Reversal of loss in surface beta-adrenoceptors was rapid after 1 hr pretreatment with isoproterenol, but followed a biphasic time course after 4 hr pretreatment, with an initial rapid return of about 40% of the down-regulated receptors, followed by a slow, gradual reappearance of receptors. The results indicate that catecholamine exposure leads to a rapid sequestration of MNL surface beta-adrenoceptors away from the cell surface, to a compartment where they are inaccessible to the hydrophilic ligand [3H]CGP-12177 as well as to the lipophilic ligand [125I]cyanopindolol at 4 degrees. Up to 2 hr of isoproterenol treatment does not lead to any breakdown of sequestered beta-adrenoceptors, as they are still recognized by [125I]cyanopindolol binding in broken cell preparations. PMID- 3036166 TI - Measurement of angiotensin converting enzyme inhibitors in serum by radioinhibitor binding displacement assay. AB - The principle of enzyme radioinhibitor binding displacement was developed to measure the concentration of angiotensin converting enzyme (ACE) inhibitors in rat serum. 125I MK351A, a tyrosyl derivative of enalaprilic acid, and a potent ACE inhibitor, bound in a concentration and time dependent manner to ACE. Binding of 125I MK351A to rat serum ACE was reduced in a concentration dependent manner in vitro by the ACE inhibitors MK521 (lisinopril), S9780, and Ro 31-3113-000 (Cilazapril diacid). This relationship was used to measure MK521 and S9780 in rat serum four hours after oral gavage with MK521, S9490-3 the prodrug ester of S9780, at 1, 2 and 4 mg/kg, or 1/2 hour after intraperitoneal injection of Ro 31 3113-000 (0.0125-0.7 mg/kg). Serum MK521 concentrations, estimated by radio inhibitor binding displacement, and radioimmunoassay, correlated well (r = 0.94, N = 9, P less than 0.001). Serum MK521, S9780 and Ro 31-3113-000 concentrations measured by radioinhibitor binding displacement assay were dose related, and inversely related to serum ACE enzymatic activity. The radioinhibitor binding displacement assay method using 125I MK351A as a ligand for ACE has application to the measurement of any competitive inhibitor of ACE. PMID- 3036167 TI - Modulation of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-mediated myelotoxicity by thyroid hormones. AB - Although binding by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) to the Ah receptor is a prerequisite for toxicity, the events responsible for subsequent TCDD effects are essentially unknown. Several lines of evidence have indicated that thyroid hormones share common molecular properties with TCDD and can modulate its toxicity. In the present studies we employed suppression of murine bone marrow hematopoiesis by TCDD as an in vitro model to study the relationship between thyroid hormones and TCDD toxicity. Supraphysiological levels of thyroid hormone mimicked TCDD myelotoxicity, in that both were inhibited by a common antagonist, 1-NH2-3,7,8-trichlorodibenzo-p-dioxin. Furthermore, myelotoxicity by both TCDD and thyroid hormone segregated with the Ah locus in congenic mice. These data provide evidence of a relationship between TCDD and thyroid hormones in that hormonal activity may help regulate TCDD toxicity. PMID- 3036168 TI - [Expressing plasmid vectors on the basis of Escherichia coli beta-galactosidase gene fragments]. AB - With the use of synthetic DNA fragments, a set of new plasmid vectors has been obtained. The vectors provided high level expression of peptides and small proteins in E. coli as fusions with fragments of beta-galactosidase of various length. These vectors were used to achieve expression of a synthetic gene for a functionally active fragment of bacteriorhodopsin. The yields of hybrid proteins consisting of beta-galactosidase and bacteriorhodopsin fragments were in the range of 5-30% from the total amount of cellular protein. PMID- 3036169 TI - [Substrate specificity of restriction endonuclease VspI]. AB - The recognition sequence and cleavage point of restriction endonuclease VspI have been determined as 5'-AT decreases TAAT. This enzyme is not isoschizomer of any known restriction endonucleases. DNA pBR322 contains a single VspI recognition sequence in position 3539. Therefore this enzyme may be used for cloning DNA in the VspI site in AmpR-gene of pBR322. PMID- 3036170 TI - [Specificity of restriction endonuclease VneI]. AB - The recognition sequence and cleavage point of restriction endonuclease VneI have been determined as 5'-G decreases TGCAC. This enzyme is not isoschizomer of any known restriction endonucleases and therefore may be widely used in investigation of DNA structure. PMID- 3036171 TI - Viral and host genetic factors influence encephalomyocarditis virus-induced polymyositis in adult mice. AB - Recent research findings implicate picornaviruses in the etiology of human polymyositis/dermatomyositis and suggest that genetic factors play a role in susceptibility to these diseases. We compared 2 variants of encephalomyocarditis (EMC) virus for their ability to induce polymyositis in adult mice, and evaluated what role the genetic background of the host plays in the degree of myositis induced. While BALB/c mice developed minimal myositis when infected with a diabetogenic variant (EMC-D), the same strain inoculated with a newly isolated myopathic variant (EMC-221A) developed viral dose-dependent elevations in muscle associated enzymes, bilateral limb muscle weakness, and the histopathologic changes of severe polymyositis. Mice with different genetic backgrounds showed significantly different susceptibilities to EMC-221A. These data suggest that the severity of polymyositis induced by EMC virus is influenced by both the viral and host genomes. PMID- 3036172 TI - Effect of soy fiber and soy protein on cholesterol metabolism and atherosclerosis in rabbits. AB - The effect of dietary fiber isolated from dehulled, defatted soybean seeds on cholesterol (CHOL) metabolism and atherosclerosis in rabbits was studied alone and in combination with isolated soy protein (ISP). Soy fiber (SF) contains both cellulosic and non-cellulosic dietary fiber. Based on the official AOAC method, soy fibers contains 75% total dietary fiber. Rabbits at 6 months of age were randomly assigned to 1 of 4 dietary treatments. All rabbits received either a casein or ISP-based diet with cellulose or SF as the only dietary fiber source for 36 weeks. Fasting blood samples and feces were collected and analyzed for lipids from individual rabbits. The entire aorta was removed and fixed, and sudanophilic stained lesions were examined visually. Rabbits consuming the SF and/or ISP diets had lower plasma CHOL levels and lower incidence of atherosclerotic lesions relative to the rabbits fed the casein-based cellulose diets. Rabbits consuming the SF and/or ISP diets also had a lower CHOL content in their liver and heart. Rabbits fed ISP-based diets had consistently increased fecal bile acid excretion, whereas rabbits fed diets containing SF had increased fecal and cholesterol concentration. These results suggest a complementary role for SF and ISP in preventing atherosclerosis in rabbits. PMID- 3036173 TI - Nuclear magnetic resonance and distance geometry studies of DNA structures in solution. PMID- 3036174 TI - Real-time spectroscopic analysis of ligand-receptor dynamics. PMID- 3036175 TI - Persistence and specificity of small doses of morphine on intake of alcoholic beverages. AB - Subsequent to water deprivation, male rats were given daily, 1.5-hr opportunities to take either water or a sweetened ethanol solution (ES). Each day, 15 min before the session, rats received a subcutaneous injection of either morphine (1.0 mg/kg) or saline. Across daily sessions, rats given saline gradually increased their intake of ES, until they were eventually taking about 2.0 to 3.0 g of ethanol/kg a session. Rats receiving morphine took greater amounts of ES from nearly the first opportunities. Additional tests assessed the effects of small doses of morphine on intakes of some sucrose solutions, and sweetened solutions containing methanol or propanol. The data support the conclusion that small doses of morphine persistently increase intake of ES across many days (up to 100) of testing, but that the effect is not unique to ES. Even though morphine's effects are not specific to ethanol, the fact that morphine persistently increases intake of ES is of interest with respect to theories of alcoholism. PMID- 3036176 TI - Selected opioids modify intake of sweetened ethanol solution among female rats. AB - Water-deprived female rats were given a daily, 1.5-hr opportunity to take either a sweetened ethanol solution or water. Across days, they increased their intake of ethanol solution and had stable intakes of about 2 g of pure ethanol/kg after 3 weeks. Morphine (1.0 mg/kg) alone, and in combination with diprenorphine (25 micrograms/kg), increased intake of ethanol solution among females similar to the increased intake seen with males under similar procedures. Fentanyl dose relatedly increased intake of ethanol. The data strengthen the idea that one or more of the endogenous opioid systems, but not all, are involved with instances of "excessive" intake of alcoholic beverages. PMID- 3036177 TI - Morphine and diprenorphine together potentiate intake of alcoholic beverages. AB - Water-deprived rats were given a daily opportunity to take water or an ethanol solution. Prior to some opportunities to drink, some were injected with morphine (across procedures either 2.0, 7.5, or 20.0 mg/kg), diprenorphine (from 0.001 to 10.0 mg/kg), or a combination of diprenorphine and morphine. The small dose of morphine increased intake of alcoholic beverage and the large dose decreased intake, confirming previous observations. Diprenorphine, across a wide range of doses, increased intake of ethanol solution. Morphine and diprenorphine together produced more intake than either given alone. Diprenorphine reversed the depressing effects of large doses of morphine on intake of ethanol solution. Since diprenorphine is an antagonist with respect to opioid analgesia and behavioral depression and an agonist with respect to intake of alcoholic beverages, and since it potentiates the small dose morphine effect, it is concluded that only some effects of morphine are related to opioid-potentiation of intake of alcoholic beverages. PMID- 3036178 TI - Anatomical localization in hippocampus of tetrahydro-beta-carboline-induced alcohol drinking in the rat. AB - Guide cannulae for unilateral or bilateral micro-injection were implanted stereotaxically into the dorsal hippocampus of the male adult Sprague-Dawley rat. Following post-operative recovery, the animal's individual preference for ethyl alcohol in concentrations from 3-30% (v/v) was tested over a 9-day period by a three-bottle, two-choice technique. Following this pre-screen, 3.0 microliter of 1,2,3,4-tetrahydro-beta-carboline (TH beta C) hydrochloride, a benzodiazepine receptor antagonist, was infused in a concentration of 25-200 ng into the hippocampus of each unrestrained rat twice a day for three to six days. After the first two days of infusion, the 9-day preference test for alcohol drinking was begun and continued identically as in the earlier test. A third alcohol preference test during which no injections were given was conducted at an interval of two weeks following the second. The micro-injection of TH beta C into certain sites in the hippocampus enhanced alcohol consumption from 0.5-2.0 g/kg during the 9-day test interval. The magnitude of this elevated intake was dependent on the site of infusion and was more pronounced when intermediate concentrations of 7-12% alcohol were offered to the rat. At sites in coronal planes encompassing AP 3.0 and AP 3.5, the micro-injection of TH beta C enhanced alcohol drinking significantly in 75% of the animals; however, when delivered at sites in coronal planes AP 1.0 through AP 2.5, TH beta C augmented alcohol drinking significantly in 15% of the rats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3036179 TI - Photoaffinity crosslinking of etorphine with opioid binding sites in the bovine adrenal medulla. AB - The covalent crosslinking of [3H]etorphine with opioid binding sites in the bovine adrenal medulla is reported. Of all the radiolabeled opiates tested (ethylketocyclazocine, etorphine, [D-Ala2, D-Leu5]enkephalin, [D-Ala2, Me-Phe4, Gly5-ol]enkephalin only etorphine could be crosslinked under uv irradiation. In our conditions (black uv lamp, 160 W, peak mean 360 nm, from a distance of 10 cm) maximum covalent binding was observed after a 10-min irradiation. Protein concentration was a crucial factor for the irreversible/total binding ratio. A good ratio (50%) was obtained at protein concentrations of about 1.0 mg/ml. Covalent binding of nonmodified opiates could be of interest for the biochemical characterization of their binding sites. PMID- 3036180 TI - Carbamate kinase of Lactobacillus buchneri NCDO110. II. Kinetic studies and reaction mechanism. AB - The participation of Mg2+ or Mn2+ nucleoside diphosphates in the reverse reaction catalyzed by purified carbamate kinase (ATP:carbamate phosphotransferase, EC 2.7.2.2) of Lactobacillus buchneri NCDO110 was studied. The results of initial velocity studies have indicated that Mn2+ ADP is as effective as a substrate as Mg2+ ADP is. Product inhibition studies have revealed that the enzyme has two distinct sites, one for nucleoside diphosphate and the other for carbamyl phosphate. The reaction of the enzyme with the substrates is of the random type. PMID- 3036181 TI - Activation of the first component of complement. PMID- 3036182 TI - Viruses perturb lymphocyte functions: selected principles characterizing virus induced immunosuppression. PMID- 3036183 TI - Lipid mediators produced through the lipoxygenase pathway. PMID- 3036184 TI - Intraoperative fine-needle aspiration and rapid diagnosis of thoracic lesions. AB - Rapid cytologic diagnosis of thoracic lesions by intraoperative fine-needle aspiration is a valuable aid to the surgeon at the time of thoracotomy or mediastinoscopy, especially for lesions which are difficult or hazardous to biopsy by standard methods. The technique is simple, quick, and accurate and aids in determining the immediate surgical management of suspicious lesions. One hundred fifty-four IOFNAs of thoracic masses performed over an 8-year period were evaluated. Diagnostic accuracy was 97.7% for the diagnosis of benign lesions, 95.4% for the diagnosis of malignancy, and 100% for the differentiation of small cell carcinoma from non-small cell carcinoma of the lung. PMID- 3036185 TI - Inorganic particulates associated with pulmonary alveolar proteinosis: SEM and X ray microanalysis results. AB - Twenty-four cases of pulmonary alveolar proteinosis (PAP) were studied by light microscopy (LM) and scanning electron microscopy (SEM) to test the hypothesis that PAP was related to silica exposure. Increased numbers of birefringent particles (vs. controls) were found in 78% of PAP cases. SEM was used to locate inorganic particulates in situ, which were individually analyzed using energy dispersive X-ray analysis. When analyzed as an aggregate group of cases, no specific inorganic particulate was evidently associated with the PAP reaction. However, analysis of individual cases revealed more specific associations. The concentration of particles determined by SEM exceeded that found by LM by a factor ranging from 2.7 to 964. The concentration of inorganic particulates per cm3 in the areas of PAP ranged from 1.3 X 10(7) to 1.02 X 10(9). Controls all had less than 10(7) particles per cm3. Available environmental history correlated well with particulate analysis results, e.g., silica in a sandblaster, metal fumes in a welder, and cement particles in a cement finisher. Particulates with unique composition were also found in cases with unavailable histories, e.g. metal fumes suggestive of welding or soldering exposure, silicates suggestive of fine particle exposure (greater than 50% of particles less than 1 micron). Only 1 case (the sandblaster) showed greater than 50% of the particles to be silica. Of the 5 infants with PAP, 3 showed the major particulate to be talc, and 1 had evidence of toxic cadmium selenide fume exposure. These results are consistent with the hypothesis that PAP, at least in the majority of cases, is associated with exposure to small inorganic particulates of several types. PMID- 3036186 TI - Neuroendocrine structures in normal and diseased human lung. AB - The increasing amount of histological, immunohistochemical and ultrastructural information on some endocrine secretions in human lung cancers suggest the need to revise the classification of neuroendocrine lesions on surgical material. The aim of the present investigation based on lung specimens removed surgically is to give further support to recent proposal for an updated classification of neuroendocrine lung carcinomas. Our study includes 58 squamous cell carcinomas, 58 adenocarcinomas, 6 large cell carcinomas, 27 neuroendocrine carcinomas, and 30 nontumourous cases. Using histological methods (HE, Alcian PAS, Grimelius silver impregnation), we illustrate the presence of neuroendocrine cells and neuroepithelial bodies with their pathological evolutions, ranging from hyperplasia, to dysplasia, and overt neoplasia. On the basis of our experience we propose the following classification of neuroendocrine carcinomas (NEC): typical carcinoids (NECNID), peripheral carcinoid or well-differentiated NEC (NECWED), NEC of intermediate or poorly differentiated type (NECINT) and NEC of small celled or microcytoma type (NECMIC). PMID- 3036187 TI - Primary malignant fibrous histiocytoma of the pleura. A case report. AB - An uncommon primary malignant fibrous histiocytoma (MFH) of the pleura is described. At light microscopy the tumor was characterized by a 'storiform' pattern of growth, with numerous multinucleated giant cells. Immunohistochemistry showed an intense, diffuse immunostaining for alpha-1-antitrypsin, focal for lysozyme. Ultrastructurally, histiocyte- and fibroblast-like cells and cells with intermediate features were present; moreover, it was possible to demonstrate the presence of mesenchymal 'stem' cells. Findings are discussed, also with respect to histogenesis of MFH. PMID- 3036188 TI - Granular cell tumor of the orbit. AB - An 18-year-old woman had a 3-month history of increasing proptosis and visual loss secondary to an inferior orbital mass. Surgical exploration revealed an encapsulated mass in the orbit. After this mass was excised good vision was restored. The diagnosis of granular cell tumor was made as a result of various microscopic studies. The location of the tumor within the orbit is unusual and its histologic origin is uncertain. Immunohistochemistry by peroxidase antiperoxidase method revealed that this tumor had S-100-protein-positive cells and negative results for lysozyme and alpha-1-antitrypsin. This fact suggests that the origin of the granular cell tumor may be due to the Schwann cell. PMID- 3036189 TI - Metastatic malignant mixed tumor of the skin. Ultrastructural and immunocytochemical characterization, histogenetic considerations and comparison with benign mixed tumors of skin and salivary glands. AB - Malignant mixed tumors of the skin (chondroid syringoma) are exceedingly rare. Few reports can be found in the literature and there are only two published cases in which electron microscopy has been performed. The precise histogenesis of this neoplasm is still disputed and the nature and mechanism of secretion of the intercellular substance is unknown as yet. Some authors believe that in mixed tumors of salivary glands this material could be produced by myoepithelial cells. A case is reported in which ultrastructural and immunocytochemical studies were performed in an effort to elucidate the histogenesis of malignant mixed tumors of skin and their relationship to other benign mixed tumors of the skin and salivary glands. The new information that has been obtained is interpreted in light of the findings provided by all published reports of malignant mixed tumors of the skin. PMID- 3036190 TI - Kallikrein in synovial fluid with rheumatoid arthritis. AB - The levels of kallikrein and collagenase in synovial fluid from rheumatoid arthritis (RA) patients were examined and the role of kallikrein in procollagenase activation is discussed. Both prekallikrein and active kallikrein in synovial fluid from patients with RA were significantly elevated when compared to synovial fluid from patients with osteoarthritis (OA). In RA synovial fluid, the ratio of the active form to total kallikrein was also higher than that in OA synovial fluid. Both active collagenase and the alpha 2-macroglobulin (alpha 2M) collagenase complex in RA synovial fluid were higher than in OA synovial fluid. A partial correlation (r = 0.58) between active kallikrein and total collagenase (active and alpha 2M-collagenase complex) was observed in RA synovial fluid. These observations indicate that both kallikrein and collagenase are associated with the destruction of cartilage, but the role of kallikrein in procollagenase activation was not fully clarified. PMID- 3036191 TI - [An autopsy case of multiple system atrophy with many Lewy bodies--striatonigral degeneration, olivo-ponto-cerebellar atrophy and autonomic nerve nucleus involvement in the spinal cord]. AB - A 72 years old man developed slowness of the motion and orthostatic hypotension at the age of 69. Neurological examination showed slight finger tremor, rigidity of extremities, bradykinesia, and marked orthostatic hypotension. The illness progressed steadily and the patient died of pneumonia. At autopsy brain weighed 1220 g. Grossly the putamen was bilaterally shrunken, the color of the substantia nigra and locus ceruleus became pale. Base of the pons and the cerebellum were atrophic. Microscopically the most remarkable change was seen in the striato nigral system. In the putamen, there were severe loss of small neurons and intense gliosis and brownish pigments were observed in the neuropil and within some of the astrocytes. There found neuronal loss and gliosis in the substantia nigra. A few Lewy bodies were seen in the substantia nigra. In the cerebellum there were slight loss of Purkinje cell and many torpedos were seen. There were demyelination and fibrirally gliosis in the cerebellar white matter except the hilus of dentate nucleus. The transverse pontocerebellar fibers were degenerated and fibrirally gliosis was seen there. The inferior olivary nuclei showed neuronal loss and astrocytosis. But the degeneration of the olivo-ponto cerebellar system in this case was not so severe as the typical case of OPCA. In the spinal cord there was depletion of nerve cells in the intermediolateral nuclei and Onufrowitz nuclei. Slight neuronal loss and many spheroids were observed in the anterior horns and there was demyelination in the corticospinal tracts.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3036192 TI - An investigation of calcium intake, 1-alpha(OH)D3, and etidronate on bone. AB - The synthetic metabolite of vitamin D3 [1 alpha(OH)D3] caused a significant plasma calcium elevation in rats only when dietary calcium was low. Animals given the low calcium diet (0.005%) had lower plasma parathyroid hormone (PTH) levels when the diet contained 1 alpha(OH)D3 and significantly higher levels than animals on a high calcium (0.95%) diet, with or without the vitamin. The nutritional stress of a low calcium diet without 1 alpha(OH)D3 resulted in a prolonged severe hypocalcemia and elevated serum PTH levels. A higher ash, phosphate, and calcium content was found in the bones of animals fed the high calcium diet, with no vitamin D3 that were given etidronate (EHDP). When animals received the same calcium diet with 1 alpha(OH)D3 supplementation, EHDP administration increased the percentage of bone ash but had no effect on ash weight. 1 alpha(OH)D3 or EHDP did not affect ash weight, dry fat free weight, and percentage of ash of bone of animals receiving a low calcium diet. The percentage of calcium and phosphorus in bone ash was similar among all groups, although the amounts per humerus were characteristically related to the calcium intake. There was approximately 20-25% less bone mineral and calcium and phosphorus in the humeri of low calcium intake animals than in animals provided an adequate dietary calcium. PMID- 3036193 TI - 'Management of the mobile fibrous ridge in the atrophic maxilla using porous hydroxyapatite blocks'. PMID- 3036194 TI - Reduction of leukotriene B4-induced intraepidermal accumulation of polymorphonuclear leukocytes by methotrexate in psoriasis. AB - The leukotriene B4 (LTB4)-induced intraepidermal accumulation of polymorphonuclear leukocytes (PMN) was quantified, using an elastase assay, in psoriatic patients receiving methotrexate and in untreated psoriatic patients. Methotrexate in therapeutic dosages inhibited PMN infiltration in six patients who had improved considerably on treatment with the drug, a gradual increase being seen during the days on which no methotrexate was taken. Two patients were in relapse, despite methotrexate treatment, and both failed to show any change in LTB4-induced accumulation of PMN. PMID- 3036195 TI - Marrow repopulation in mice treated with busulphan or isopropyl methane sulphonate and bone marrow. AB - By using karyotypic analysis of female mice treated with busulphan or isopropyl methane sulphonate (IMS), and injected with male bone marrow the donor contribution to both total marrow cellularity and spleen colony forming cells (CFU-S) was assessed for up to 6 months after transplant. In the mice treated with busulphan the marrow cells yielded metaphases of which between 40% and 83% were of donor type. Between 60% and 97% of metaphases in spleen colonies formed in irradiated mice were of donor type during the 24-week study period. In contrast, mice prepared for the transplant with IMS showed no cells of donor type at any time after transplant, neither did they possess CFU-S of donor type. We were therefore led to conclude that the donor cells made no contribution to longterm engraftment in mice prepared with IMS, whilst in those prepared with busulphan they were the predominantly active haemopoietic cells. These results are consistent with a model of haemopoiesis in which the most primitive cells reside in a 'niche' where they are resistant to the effects of IMS but susceptible to the action of busulphan. Busulphan may vacate some niches to allow engraftment by transplanted marrow, whilst IMS yields no unoccupied niches for grafted cells to occupy, and cannot therefore lead to a stable chimaerism. PMID- 3036196 TI - Pyrimidine nucleoside monophosphate kinase hyperactivity in hereditary erythrocyte pyrimidine 5'-nucleotidase deficiency. AB - The pyrimidine nucleoside triphosphates (CTP, UTP) increase in the pyrimidine 5' nucleotidase (P5N) deficient red blood cell (RBC) to a greater degree than do the pyrimidine nucleoside monophosphates (CMP, UMP). Pyrimidine nucleoside monophosphate (PNMP) kinase phosphorylates CMP and UMP to their respective phosphodiesters. We tested the hypothesis that increased PNMP kinase activity contributes to the disproportionate increase in CTP and UTP in the P5N deficient RBC. CMP and UMP kinase activities were increased in high reticulocyte (4.4 +/- 2.1 and 8.5 +/- 3.3 mumol/ml RBC per minute) compared to normal RBC (2.8 +/- 1.0 and 6.0 +/- 2.5 mumol/ml RBC per minute). P5N deficient RBC (n = 2) had significantly increased CMP and UMP kinase activities (14.0 and 26.5 mumol/ml RBC per minute). UMP and CDP-ethanolamine were able to increase the activity of CMP kinase in crude haemolysate and the activity of partially purified enzyme. Since the Km for CMP of CMP kinase was 33 mumol/l in P5N deficient RBC and since the CMP concentration is 25-90 mumol/l in the P5N deficient RBC, the enzyme should be nearly saturated with CMP in the P5N deficient RBC. Thus, PNMP kinase hyperactivity appears to contribute to the disproportionate increase in CTP and UTP in the P5N deficient RBC. PMID- 3036197 TI - Relative nutritional availability to rats of selenium in Finnish spring wheat (Triticum aestivum L.) fertilized or sprayed with sodium selenate and in an American winter bread wheat naturally high in Se. AB - A Finnish national programme to fertilize crops with sodium selenate led us to compare the nutritional availability to rats of selenium in two Finnish spring wheats (Triticum aestivum L.), either fertilized or sprayed with sodium selenate, with that in an American winter bread wheat naturally high in Se. Weanling male rats were given a Se-deficient Torula yeast diet for 4 weeks followed by either continued depletion or repletion for 4 weeks with graded levels of Se as sodium selenite (standard) or wheat (test food). Plasma and liver Se levels and plasma and liver glutathione peroxidase (EC 1.11.1.9; GSH-Px) activities were used as criteria of body Se status. The availability of Se under these conditions was calculated with the point-slope technique at two dietary levels of Se (Expt 1) and with the slope-ratio method (Expt 2). In the point-slope assay, the level of dietary Se fed had a considerable effect on the apparent availability values obtained which made interpretation of the results difficult. In the slope-ratio assay, no difference in the availability of Se from the various wheats was observed when plasma or liver Se levels were used as the response criteria. The Se in the fertilized wheat was somewhat more available than that in the sprayed wheat when plasma or liver GSH-Px activities were the response criteria. Overall, availability values (%) derived by averaging all four response criteria were 86, 77 and 73 for the fertilized and sprayed Finnish wheats and the American wheat respectively (sodium selenite 100). These results show that wheat is a relatively available source of Se to rats regardless of whether its Se content is naturally high or is increased by fertilization or spraying. PMID- 3036198 TI - The role of rumen protozoa in the utilization of paspalum (Paspalum dilatatum) hay by cattle. AB - Six Friesian heifers (250 kg live weight) with permanent cannulas in the rumen and abomasum were allocated at random into two groups of three. One group was treated with Teric GN9 (ICI (Aust.) Ltd) to defaunate the animals during the first two of the four periods of the experiment, after which they were refaunated. The second group was treated with Teric at the end of the first two periods. The dietary treatments were: paspalum (Paspalum dilatatum) hay (4.1 kg/d) given alone and the hay supplemented with urea (20 g/kg dry matter). Defaunation was not complete but the approximate volume of protozoa in the rumen of treated animals was less than 6% of that in the untreated animals. The amount of organic matter (OM) digested in the stomach was lower (P less than 0.01) in animals with reduced fauna than in those with normal fauna. There were reductions in both the apparent OM digestibility in the total tract (from 0.56 to 0.52, P less than 0.01) and the proportion of the digestible OM digested in the rumen (from 0.82 to 0.79, not significant) of animals with reduced fauna. Apparent digestibilities of acid-detergent fibre and neutral-detergent fibre were significantly lower (P less than 0.01) in animals with reduced fauna. The amount of nitrogen disappearing from the stomach was significantly higher (P less than 0.01) with the urea supplement; effects due to concentrations of protozoa were not significant. The flow of non-ammonia-N from the abomasum was higher (P less than 0.05) in animals with reduced fauna than in animals with normal fauna. The flows of bacterial N from the abomasum and the efficiencies of bacterial N synthesis were not significantly affected by the treatments. N retention was higher (P less than 0.01) in animals receiving the urea supplement but differences due to protozoa were not significant. Protozoal contribution to the microbial N flowing from the rumen of animals with normal fauna was estimated to be 24 and 27% with and without the urea supplement respectively. Concentrations of rumen-fluid ammonia-N were reduced (P less than 0.05) and those of volatile fatty acids were increased (P less than 0.01) with reduction in protozoal numbers. Molar proportions of propionic acid increased (P less than 0.05) and of butyric acid decreased (P less than 0.01) with reduced rumen fauna.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3036199 TI - Use of particle-bound microbial enzyme activity to predict the rate and extent of fibre degradation in the rumen. AB - A method was developed for extracting enzymes from micro-organisms closely associated with ammonia-treated straw (NH3-S) that had been incubated in nylon bags in the rumen. Incubation of washed straw with 125 ml carbon tetrachloride/l and 20 micrograms lysozyme/ml for 3 h at 37 degrees gave carboxymethylcellulase (EC 3.2.1.4; CMCase) and NAD-linked glutamate dehydrogenase (EC 1.4.1.2; GDH) activities greater than those extracted by sonication. GDH associated with NH3-S increased with incubation time and was highest in sheep receiving a high-barley diet. Particle-bound CMCase activity reached a peak between 16 and 24 h and declined thereafter. Particle-bound GDH activity showed no correlation with dry matter (DM) degradation in the rumens of sheep fed on a range of diets. In contrast, CMCase activity after 24 h was highly correlated with DM degradability of the same samples at 24 h (r 0.98) and 48 h (r 0.94). It was concluded that GDH and CMCase can be used as indices of the total population of colonizing rumen micro-organisms and of the fibre-degrading population respectively, and that these enzymes can therefore be used to assess rapidly and with great sensitivity variations in the rumen environment that affect the rate of fibre breakdown. PMID- 3036200 TI - Effect of fasting and of methionine deficiency on L-methionine, DL-methionine and DL-2-hydroxy-4-methylthiobutanoic acid metabolism in broiler chicks. AB - Metabolism of L-[1-14C]methionine, DL-[1-14C]methionine and DL-[1-14C]2-hydroxy-4 methylthiobutanoic acid (DL-HMB) by broiler chicks which had been fasted overnight or given a methionine-deficient diet was compared with fed (control) birds. The excretion of 14C-labelled material, total 14CO2 exhaled, 14C incorporation into tissue proteins and the 14C-labelled material in perchloric acid-soluble tissue fractions were measured 6 h after injection of the 14C labelled materials. The incorporation of 14C into tissue proteins and the relative rates of conversion of D-methionine and DL-HMB to L-methionine in tissues under different nutritional regimens were compared using protein-bound 14C:protein-free 14C values. Fasted birds exhaled more 14CO2 than control birds but excreted less 14C, while methionine-deficient birds behaved very similarly to the control animals in these respects. Fasted birds incorporated much less 14C into proteins of tissues other than liver and kidney from all three labelled tracers. The values for protein-bound 14C:protein-free 14C were lower in all tissues. Methionine-deficient birds had similar levels of 14C in tissue proteins but lower values for protein bound 14C:protein-free 14C. Examination of the values for protein-bound 14C:protein-free 14C suggest that brain and probably liver tissues from fasted and methionine-deficient birds showed improved rates of conversion of D-methionine and DL-HMB to L-methionine compared with control animals. PMID- 3036201 TI - Porto-arterial plasma concentration differences of urea and ammonia-nitrogen in growing pigs given high- and low-fibre diets. AB - The effects of a high- (HF) and a low- (LF) fibre diet on porto-arterial plasma concentration differences and plasma levels of urea ammonia-nitrogen were compared in six Swedish Landrace X Yorkshire pigs (30-52 kg). Portal and arterial blood samples were drawn during 8 h following two consecutive meals, given at 08.00 and 16.00 hours. The HF diet, in comparison with the LF diet, was found to produce significantly lower portal plasma urea levels. However, concomitant arterial plasma were equally depressed, consequently leaving the porto-arterial urea differences unchanged. In no instance during the 16 h studied could negative porto-arterial urea differences clearly be observed. It was thus concluded that none of the diets induced an important net flux of plasma urea directly from the circulation into the gastrointestinal tract. The significantly lower circulating plasma urea levels that were observed when the pigs received the HF diet were also associated with significantly lower urinary excretions of urea. The HF and LF diets had similar effects on portal and arterial plasma levels of ammonia-N. This was also true with regard to porto-arterial ammonia-N differences. PMID- 3036202 TI - Crystal structure of cytochrome c peroxidase compound I. AB - We have compared the 2.5-A crystal structure of yeast cytochrome c peroxidase (CCP) with that of its semistable two-equivalent oxidized intermediate, compound I, by difference Fourier and least-squares refinement methods. Both structures were observed at -15 degrees C. The difference Fourier map reveals that formation of compound I causes only small positional adjustments of a few tenths of an angstrom. The map's most pronounced feature is a pair of positive and negative peaks bracketing the heme iron position. Least-squares refinement shows that the iron atom moves about 0.2 A toward the distal side of the heme. No significant difference density is evident near the side chains of Trp-51 or Met-172, each of which has been proposed to be the site of the electron paramagnetic resonance (EPR) active radical in compound I. However, the second most prominent feature of difference density is a negative peak near the side chain of Thr-180, which, according to the results of least-squares refinement, moves by 0.15 A in the direction of Met-230. These observations, together with the results of mutagenesis experiments [Fishel, L. A., Villafranca, J. E., Mauro, J. M., & Kraut, J. (1987) Biochemistry 26, 351-360; Goodin, D. B., Mauk, A. G., & Smith, M. (1986) Proc. Natl. Acad. Sci. U.S.A. 83, 1295-1299] in which Trp-51 and Met 172 have been replaced without loss of the EPR radical signal in compound I, lead us to consider the possibility that the radical site lies within a cluster composed of the side chains of Met-230, Met-231, and Trp-191.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3036203 TI - Interactions of T7 RNA polymerase with T7 late promoters measured by footprinting with methidiumpropyl-EDTA-iron(II). AB - The interactions of T7 RNA polymerase with T7 late promoters were studied by using quantitative footprinting with methidiumpropyl-EDTA X Fe(II) [MPE-Fe(II)] as the DNA cleaving agent. Class II and class III T7 promoters have a highly conserved 23 base pair sequence from -17 to +6. Among class III promoters the -22 to -18 region is also highly conserved. For a class II promoter, T7 RNA polymerase protects the -17 to -4 region from MPE-Fe(II) cleavage; when GTP is present, protection extends from -17 to +5 (noncoding strand). For a class III promoter, protection extends from -20 to -4 and in the presence of GTP from -20 to +5 (noncoding strand). The protected regions for the coding strands of both promoters were nearly identical with that seen for the noncoding strands. The binding constant for the class III promoter is (4 +/- 1.5) X 10(7) M-1 and in the presence of GTP increases to (10 +/- 1.7) X 10(7) M-1. These binding constants are about 1000 and 200 times greater, respectively, than values reported previously [Ikeda, R. A., & Richardson, C. C. (1986) Proc. Natl. Acad. Sci. U.S.A. 83, 3614-3618]. The differences in binding constants are probably due to tRNA and high salt used in those earlier experiments. Both tRNA and high salt (greater than 50 mM NaCl and greater than 10 mM MgCl2) inhibit the binding of the polymerase to the promoter. Optimal binding conditions occur at 2-5 mM MgCl2 and 0-10 mM NaCl.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3036204 TI - Ascorbate depletion as a consequence of product recycling during dopamine beta monooxygenase catalyzed selenoxidation. AB - The competence of dopamine beta-monooxygenase (DBM) to process selenide substrates was investigated, in anticipation that the expected selenoxide products would exhibit unique reactivity and redox properties. The prototypical selenide phenyl 2-aminoethyl selenide (PAESe) was synthesized and shown to be a substrate for DBM with the characteristic e/O2 ratio of 2:1 for monooxygenation. The kinetic parameters for oxygenation of PAESe were found to be similar to those for the DBM-catalyzed sulfoxidation of the cognate sulfide phenyl 2-aminoethyl sulfide [May, S. W., & Phillips, R. S. (1980) J. Am. Chem. Soc. 102, 5981-5983], and selenoxidation was stimulated by fumarate in a manner similar to other well characterized DBM monooxygenation reactions. Identification of phenyl 2 aminoethyl selenoxide (PAESeO) as the enzymatic product was accomplished by the demonstration of coincident elution of authentic PAESeO with the enzymatic product in three significantly different HPLC systems. PAESeO was found to oxidize ascorbic acid with the concomitant and stoichiometric reduction of PAESeO back to the selenide, PAESe. As a consequence of this nonenzymatic reaction, ascorbate-supported DBM turnover was prematurely terminated under standard assay conditions due to depletion of reduced ascorbate. The kinetics of the redox reaction between PAESeO and ascorbate were investigated with a spectrophotometric assay of ascorbate at 300 nm, and a second-order rate constant of 3.4 M-1 s-1 was determined at pH 5.0, 25 degrees C. Spectrophotometric assay of cytochrome c (cyt c) reduction at 550 nm during the oxidation of ascorbate by PAESeO demonstrated that no cyt c trappable semidehydroascorbate was produced in this nonenzymatic reaction.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3036205 TI - NMR studies of the AMP-binding site and mechanism of adenylate kinase. AB - NMR has previously been used to determine the conformation of enzyme-bound MgATP and to locate the MgATP-binding site on adenylate kinase [Fry, D. C., Kuby, S. A., & Mildvan, A. S. (1985) Biochemistry 24, 4680-4694]. To determine the conformation and location of the other substrate, AMP, distances have been measured from Cr3+AMPPCP, a linear competitive inhibitor with respect to MgATP, to six protons and to the phosphorus atom of AMP on adenylate kinase, with the paramagnetic probe-T1 method. Time-dependent nuclear Overhauser effects (NOEs) have been used to measure five interproton distances on enzyme-bound AMP. These distances were used to determine the conformation of bound AMP in addition to its position with respect to metal-ATP. Enzyme-bound AMP exhibits a high anti glycosyl torsional angle (chi = 110 +/- 10 degrees), a 3'-endo,2'-exo ribose pucker (delta = 105 +/- 10 degrees), and gauche-trans orientations about the C4' C5' bond (gamma = 180 +/- 10 degrees) and the C5'-O5' bond (beta = 170 +/- 20 degrees). The distance from Cr3+ to the phosphorus of AMP is 5.9 +/- 0.3 A, indicating a reaction coordinate distance of approximately 3 A, which is consistent with an associative SN2 mechanism for the phosphoryl transfer. Ten intermolecular NOEs, from protons of the enzyme to those of AMP, were detected, indicating the proximity of at least three hydrophobic amino acids to bound AMP. These constraints, together with the conformation of AMP and the intersubstrate distances, were used to position AMP into the X-ray structure of adenylate kinase. The AMP binding site is found to be near (less than or equal to 4 A from) Leu-116, Arg-171, Val-173, Val-182, and Leu-190; all of these residues have been found to be invariant in muscle-type rabbit, calf, human, porcine [Kuby, S. A., Palmieri, R. H., Frischat, A., Fischer, A. H., Wu, L. H., Maland, L., & Manship, M. (1984) Biochemistry 23, 2393-2399], and chicken adenylate kinase [Kishi, F., Maruyama, M., Tanizawa, Y., & Nakazawa, A. (1986) J. Biol. Chem. 261, 2942-2945]. PMID- 3036206 TI - Effect of single amino acid changes in the region of the adenylylation site of T4 RNA ligase. AB - Preparation and analysis of a series of mutants of bacteriophage T4 RNA ligase that carry single amino acid changes at or near the site of covalent reaction with ATP (adenylylation) are described. The mutant proteins were constructed by site-directed mutagenesis of the gene for T4 RNA ligase (g63) cloned in M13 vectors, transfer of the mutant genes into a lambda pL-containing expression plasmid, and subsequent expression in Escherichia coli. The results give further evidence that Lys-99 is the adenylylation site and that the residue is also important to step 3 in the RNA ligase mechanism (ligation between acceptor and adenylylated donor). Mutations at Glu-100 or Asp-101 have no effect on adenylylation, but Asp-101 is shown to be crucial to both step 2 (transfer of adenylyl to donor) and step 3. PMID- 3036208 TI - Inactivation of Escherichia coli glycerol kinase by 5'-[p (fluorosulfonyl)benzoyl]adenosine: protection by the hydrolyzed reagent. AB - Incubation of Escherichia coli glycerol kinase (EC 2.7.1.30; ATP:glycerol 3 phosphotransferase) with 5'-[p-(fluorosulfonyl)benzoyl]adenosine (FSO2BzAdo) at pH 8.0 and 25 degrees C results in the loss of enzyme activity, which is not restored by the addition of beta-mercaptoethanol or dithiothreitol. The FSO2BzAdo concentration dependence of the inactivation kinetics is described by a mechanism that includes the equilibrium binding of the reagent to the enzyme prior to a first-order inactivation reaction in addition to effects of reagent hydrolysis. The hydrolysis of the reagent has two effects on the observed kinetics. The first effect is deviation from pseudo-first-order kinetic behavior due to depletion of the reagent. The second effect is the novel protection of the enzyme from inactivation due to binding of the sulfonate hydrolysis product. The rate constant for the hydrolysis reaction, determined independently from the kinetics of F- release, is 0.021 min-1 under these conditions. Determinations of the reaction stoichiometry with 3H-labeled FSO2BzAdo show that the inactivation is associated with the covalent incorporation of 1.08 mol of reagent/mol of enzyme subunit. Ligand protection experiments show that ATP, AMP, dAMP, NADH, 5' adenylyl imidodiphosphate, and the sulfonate hydrolysis product of FSO2BzAdo provide protection from inactivation. The protection obtained with ATP is not dependent on Mg2+. Less protection is obtained with glycerol, GMP, etheno-AMP, and cAMP. No protection is obtained with CMP, UMP, TMP, etheno-CMP, GTP, or fructose 1,6-bisphosphate. The results are consistent with modification by FSO2BzAdo of a single adenine nucleotide binding site per enzyme subunit. PMID- 3036207 TI - Characterization and sequence-specific binding to mouse mammary tumor virus DNA of purified activated human glucocorticoid receptor. AB - Activated glucocorticoid receptor (GR) from the human cell line HeLa S3 was purified by differential chromatography on DNA-cellulose followed by DEAE Sepharose chromatography to 50-60% homogeneity according to sodium dodecyl sulfate gel electrophoresis and densitometric scanning of silver-stained gels. These gels routinely demonstrated a main band of Mr 94,000 (94K band) and two minor bands of Mr 79,000 (79K band) and 39,000 (39K band), respectively. Photoaffinity labeling indicated that the hormone was bound to the 94K and 79K components. In some preparations, a 72K band was observed. Further characterization of the purified receptor by gel permeation chromatography on Sephadex G-200 revealed a receptor complex with a Stokes radius of 5.8 nm. The sedimentation coefficient of the purified receptor was 4.4 Sw. In analogy to the rat hepatic GR, limited proteolysis of the purified GR with trypsin or alpha chymotrypsin led to degradation of the 94K and 79K components and appearance of 28K and 39K fragments, respectively. In addition, no difference in the protease digestion pattern using Staphylococcus aureus V8 protease was observed. Immunoblotting using a monoclonal antibody raised against the 94K GR from rat liver demonstrated cross-reactivity with the human 94K and 79K proteins from HeLa S3 cells, indicating similar antigenic characteristics between rat and human GR. In our study, five out of nine tested monoclonal antibodies against the rat liver GR cross-reacted with human GR. DNase I and exonuclease III protection experiments demonstrated binding of the purified human GR to specific GR binding regions in mouse mammary tumor virus DNA.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3036209 TI - Isotope effect evidence for the zinc hydroxide mechanism of carbonic anhydrase catalysis. AB - The carbon kinetic isotope effect on the enzymatic dehydration of HCO3- ion is k12/k13 = 1.011 and is independent, within experimental error, of the addition of sucrose, substitution of D2O for H2O, and substitution of enzyme-bound Zn2+ by Co2+. These results are consistent with a ping-pong mechanism in which proton transfer between enzyme and solvent is separated from HCO3- dehydration. For the dehydration half-reaction, diffusional processes are severalfold faster than dehydration, and the rate-determining step is the dehydration itself. The intrinsic isotope effect is approximately 1.011, indicating that hydration of CO2 occurs by reaction of zinc-bound OH-, rather than zinc-bound H2O. PMID- 3036210 TI - Influence of the calcium-induced gel phase on the behavior of small molecules in phosphatidylserine and phosphatidylserine-phosphatidylcholine multilamellar vesicles. AB - The behavior of fluorescent and spin-label probes is examined in several fluid and gel phospholipid phases, with particular focus on the Ca2+-induced gel phase in phosphatidylserine (PS). These probes have behavior characteristic of the type of probe and of the type of lipid environment. Anthroyloxy- and doxyl-labeled PS [12-AS-PS and (7,6)PS, respectively] exhibit greatly restricted and/or slow probe motion in Ca(PS)2, even compared to thermotropic gel-phase lipid at the same temperature. In contrast, anthroyloxy- and doxyl-labeled phosphatidylcholine (PC), as well as fluorescent-labeled and spin-labeled fatty acid derivatives, show no apparent change in probe motion in Ca(PS)2 compared to fluid lamellar lipid. Doxyl-labeled phosphatidic acid, phosphatidylethanolamine, and phosphatidylglycerol show restricted motion in Ca(PS)2 relative to fluid-phase lipid, but the electron paramagnetic resonance (EPR) spectra could not be interpreted in terms of simple models for probe ordering. The fluorescent probes diphenylhexatriene (DPH) and trans-parinaric acid methyl ester (tPNA-Me) show motional behavior in Ca(PS)2 that is intermediate between that observed in fluid and in thermotropic gel-phase lipid. When Ca(PS)2 and fluid PS/PC phases coexist, probe molecules distribute between the two phases. Experiments using fluorescence quenching by spin-labeled PC in PS/PC in excess Ca2+ yield the distribution of several fluorophore probes between fluid liquid-crystal and Ca(PS)2 gel phases, expressed as a concentration ratio, RLC/G. The value of RLC/G = 100 in favor of the fluid phase is obtained for 12-AS-PC, 18 for 12-AS-Me, 12 for DPH, 3 for tPnA Me, and 1 for 12-AS-PS.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3036212 TI - Saturation-transfer electron paramagnetic resonance detection of anisotropic motion by sickle hemoglobin molecules in the polymer state. AB - The motional behavior of spin-labeled deoxygenated sickle hemoglobin has been studied by using both 9- and 35-GHz saturation-transfer electron paramagnetic resonance (EPR). Using spectral subtraction techniques and saturation-transfer EPR parameter correlation plots, we find that the saturation-transfer EPR spectra for the sickle hemoglobin gel state at high temperature and high hemoglobin concentration cannot be described as a simple superposition of spectra from immobilized hemoglobin plus solution-state hemoglobin but instead suggest that the individual sickle hemoglobin molecules exhibit limited, anisotropic, rotational oscillation within the polymer fiber. The spectra also imply that the symmetry axis for sickle hemoglobin rotational oscillation is approximately coincident with the nitroxide z axis of the covalently attached spin-label. We suggest that this anisotropic rotational motion may be produced by one or two of the known intermolecular contact sites within the sickle hemoglobin fiber acting as strong intermolecular binding sites, and producing "motional alignment" within the fiber; determining the location of the strong binding site should be important in focusing the future development of antisickling agents. PMID- 3036211 TI - Covalent labeling of a high-affinity, guanyl nucleotide sensitive parathyroid hormone receptor in canine renal cortex. AB - Putative parathyroid hormone (PTH) receptors in canine renal membranes were affinity labeled with 125I-bPTH(1-34) using the heterobifunctional cross-linking reagent N-hydroxysuccinimidyl 4-azidobenzoate. Sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed the presence of a major 85,000 molecular weight (Mr) PTH binding component, the labeling of which was inhibited by nanomolar concentrations of unlabeled PTH and by micromolar concentrations of 5'-guanylyl imidodiphosphate [Gpp-(NH)p]. Labeling was not influenced by the unrelated peptides insulin and arginine vasopressin. Minor PTH binding components of Mr 55,000 and 130,000 were also seen, and labeling of these was likewise sensitive to unlabeled PTH and to Gpp(NH)p. Omission of protease inhibitors during the isolation of plasma membranes resulted in the loss of the Mr 85,000 PTH binding species and the appearance of an Mr 70,000 form. Several minor PTH binding components also were observed. Equilibrium binding studies showed that such membranes had an affinity for PTH indistinguishable from that in membranes isolated with protease inhibitors and displaying a major Mr 85,000 PTH binding species. We conclude that the major form of the adenylate cyclase coupled PTH receptor in canine renal membranes is an Mr 85,000 protein. An endogenous enzyme, probably a lysosomal cathepsin, can cleave this form to produce an Mr 70,000 receptor that retains full functional activity with respect to high-affinity, guanyl nucleotide sensitive PTH binding. The ability to covalently label the PTH receptor in high yield represents a major step toward the structural characterization of this important detector molecule. PMID- 3036213 TI - Structure of the class II enzyme of human liver alcohol dehydrogenase: combined cDNA and protein sequence determination of the pi subunit. AB - The class II enzyme of human liver alcohol dehydrogenase was isolated, carboxymethylated, and cleaved with CNBr and proteolytic enzymes. Sequence analysis of peptides established structures corresponding to the pi subunit. Two segments from the C-terminal region unique to pi were selected for synthesis of oligodeoxyribonucleotide probes to screen a human liver cDNA library constructed in plasmid pT4. Sequence analysis of two identical hybridization-positive clones with cDNA inserts of about 2000 nucleotides gave the entire coding region of the pi subunit, a 61-nucleotide 5' noncoding region and a 741-nucleotide 3' noncoding region containing four possible polyadenylation sites. Translation of the coding region yields a 391-residue polypeptide, which in all regions except the C terminal segment corresponds to the protein structure as determined directly by peptide analysis. With the class I numbering system, the exception concerns a residue exchange at position 368, the actual C-terminus which is Phe-374 by peptide data but a 12-residue extension by cDNA data, and possibly two further residue exchanges at positions 303 and 312. The size difference might indicate the existence of posttranslational modifications of the mature protein or, in combination with the residue exchanges, the existence of polymorphism at the locus for class II subunits. The pi subunit analyzed directly results in a 379 residue polypeptide and is the only class II size thus far known to occur in the mature protein.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3036214 TI - Preparation and properties of cardiac cytochrome c1. AB - A method for the large-scale isolation of beef heart mitochondrial cytochrome c1 in high purity was developed. This method gave higher yield of "one-band" cytochrome c1 than previously reported [Kim, C. H., & King, T. E. (1981) Biochem. Biophys. Res. Commun. 102, 607-614]. In addition, the present method was effective in the preparation of "two-band" cytochrome c1 which was used to prepare the hinge protein according to the principle of sequential resolution [Kim, C. H., & King, T. E. (1983) J. Biol. Chem. 258, 13543-13551]. The isolation of one-band and two-band cytochrome c1 by this procedure could be completed within 3 or 4 days starting with succinate-cytochrome c reductase. One-band cytochrome c1 showed a molecular weight of 44,000 by sedimentation equilibrium and 29,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The disparities in these data from the actual value of 27,924 by amino acid sequence analysis, as previously reported [Wakabayashi, S., Matsubara, H., Kim, C. H., & King, T. E. (1982) J. Biol. Chem. 257, 9335-9344], are most probably due to the formation of detergent or detergent-phosphate complex. A comparison of some properties of one-band cytochrome c1 with those of two-band cytochrome c1 clearly showed significant differences between the two preparations. These results suggest the hypothesis that one of the possible roles of the hinge protein in the mitochondrial respiratory chain is to stabilize the conformation of cytochrome c1. PMID- 3036215 TI - Phosphonamidate inhibitors of human neutrophil collagenase. AB - A series of phosphonamidates has been synthesized and shown to inhibit human neutrophil collagenase. The compounds all have sequences patterned after the cleavage site in the alpha 1(I) chain of type I collagen, except that the carbonyl group of the Gly residue in subsite P1 has been replaced by a P(= O)(OH) group (abbreviated GlyP). As the central GlyP-Leu unit is lengthened in the N- and C-terminal directions, in accordance with the cleavage sequence found in collagen, inhibition is systematically improved. The best inhibitor is Cbz-GlyP Leu-Ala-Gly, which inhibits competitively with a KI value of 14 microM. These phosphonamidates are thought to be acting as transition-state analogues. PMID- 3036216 TI - ADP-ribosyl transferase and NAD glycohydrolase activities in rat liver mitochondria. AB - ADP-ribosyl transferase and NAD glycohydrolase activities have been estimated in mitochondria in mitoplasts as well as in other submitochondrial fractions. A high activity of these two enzymes was present in mitoplasts as compared to the outer membrane preparation or intermembrane compartment. Inhibitor studies provide strong evidence for the involvement of ADP-ribosyl transferase in the process of ADP-ribosylation of mitochondrial proteins. When NAD glycohydrolase was blocked by nicotinamide or 3-aminobenzamide, the incorporation of ADP-ribose into mitochondrial proteins still occurs. ADP-ribosyl transferase activity could also be detected when NAD glycohydrolase was separated by hydroxylapatite chromatography. The protein-linked ADP-ribose moiety appears to be an oligomer in mitochondria. PMID- 3036217 TI - Adenosine kinase from human erythrocytes: kinetic studies and characterization of adenosine binding sites. AB - The reaction catalyzed by adenosine kinase purified from human erythrocytes proceeds via a classical ordered sequential mechanism in which adenosine is the first substrate to bind to and AMP is the last product to dissociate from the enzyme. However, the interpretation of the steady-state kinetic data is complicated by the finding that while AMP acts as a classical product inhibitor at concentrations greater than 5 mM, at lower concentrations AMP can act as an apparent activator of the enzyme under certain conditions. This apparent activation by AMP is proposed to be due to AMP allowing the enzyme mechanism to proceed via an alternative reaction pathway that avoids substrate inhibition by adenosine. Quantitative studies of the protection of the enzyme afforded by adenosine against both spontaneous and 5,5'-dithiobis(2-nitrobenzoic acid) mediated oxidation of thiol groups yielded "protection" constants (equivalent to enzyme-adenosine dissociation constant) of 12.8 microM and 12.6 microM, respectively, values that are more than an order of magnitude greater than the dissociation constant (Kia = 0.53 microM) for the "catalytic" enzyme-adenosine complex. These results suggest that adenosine kinase has at least two adenosine binding sites, one at the catalytic center and another quite distinct site at which binding of adenosine protects the reactive thiol group(s). This "protection" site appears to be separate from the nucleoside triphosphate binding site, and it also appears to be the site that is responsible for the substrate inhibition caused by adenosine. PMID- 3036218 TI - Structural and biochemical properties of bidentate tetraaquarhodium(III) complexes of inorganic pyrophosphate and adenosine 5'-diphosphate. AB - The structural and biochemical properties of the alpha,beta-bidentate tetraaquarhodium(III) complexes of inorganic pyrophosphate [Rh(H2O)4PP] and adenosine diphosphate [Rh(H2O)4ADP] are examined. These Rh(III) complexes are exchange-inert analogues of the corresponding physiologically important MgIIPP and MgIIADP complexes. The crystal structure of [Rh(H2O)4H2P2O7]+Cl- shows that the six-membered chelate ring adopts a twist-boat conformation with an unusually high puckering amplitude of 0.756 (3) A. The Rh coordination distances average 2.02 (1) A, while the bridge P-O bonds are virtually equal in length. All 10 protons of the complex participate in hydrogen bonding. There are two intramolecular hydrogen bonds between the phosphate oxygen atoms and the axially coordinated water molecules. The Rh(H2O)4PP complex was found to be a substrate for yeast inorganic pyrophosphatase, with Ki = 0.063 (7) mM and Vm = 500 (100) min-1. The two screw sense isomers of Rh(H2O)4ADP were prepared from (Rp)-[alpha 16O,18O]ADP and assigned configuration on the basis of the magnitude of their 31P NMR isotopic chemical shifts. The Rh(H2O)4ADP complex binds a number of kinases as tightly as MgADP. Arginine kinase and creatine kinase were shown to bind the delta Rh(H2O)4ADP isomer 7 and 45 times tighter, respectively, than the lambda isomer. The reactivity of Rh(H2O)4PP with pyrophosphatase is comparable to that of Cr(H2O)4PP, and the binding affinities of the Rh(H2O)4ADP screw sense isomers for kinases are also comparable to those observed for the corresponding Cr(H2O)4ADP screw sense isomers. PMID- 3036219 TI - Resolution of the circular dichroism spectra of the mitochondrial cytochrome bc1 complex. AB - The circular dichroic spectrum of the mitochondrial cytochrome bc1 complex isolated from bovine heart has been resolved into the contributions from the prosthetic groups: cytochrome c1, the 'Rieske' iron-sulphur centre and the two b cytochromes. It is apparent that firstly, the circular dichroism (CD) properties of cytochrome c1 within the bc1 complex differ from those found in the isolated cytochrome c1 and secondly, both the oxidized and reduced b cytochromes exhibit an intense spectrum of bilobic shape, with the wavelengths of the cross-over points closely corresponding to those of the maxima in the optical absorbance spectra. These latter CD features are discussed in relation to the proposed structure of cytochrome b. PMID- 3036220 TI - Effects of adriamycin on respiratory chain activities in mitochondria from rat liver, rat heart and bovine heart. Evidence for a preferential inhibition of complex III and IV. AB - The inhibition of respiratory chain activities in rat liver, rat heart and bovine heart mitochondria by the anthracycline antibiotic adriamycin was measured in order to determine the adriamycin-sensitive sites. It appeared that complex III and IV are efficiently affected such that their activities were reduced to 50% of control values at 175 +/- 25 microM adriamycin. Complex I displayed a minor sensitivity to the drug. Of the complex-I-related activities tested, only duroquinone oxidation appeared sensitive (50% inhibition at approx. 450 microM adriamycin). Electron-transfer activities catalyzed by complex II remained essentially unaltered up to high drug concentrations. Of the activities measured for this complex, only duroquinone oxidation was significantly affected. However, the adriamycin concentration required to reduce this activity to 50% exceeded 1 mM. Mitochondria isolated from rat liver, rat heart and bovine heart behaved essentially identical in their response to adriamycin. These data support the conclusion that, in these three mitochondrial systems, the major drug-sensitive sites lie in complex III and IV. Cytochrome c oxidase and succinate oxidase activity in whole mitochondria exhibited a similar sensitivity towards adriamycin, as inner membrane ghosts, suggesting that the drug has direct access to its inner membrane target sites irrespective of the presence of the outer membrane. By measuring NADH and succinate oxidase activities in the presence of exogenously added cytochrome c, it appeared that adriamycin was less inhibitory under these conditions. This suggests that adriamycin competes with cytochrome c for binding to the same site on the inner membrane, presumably cardiolipin. PMID- 3036221 TI - Role of de novo protein synthesis in the interconversion of glucose transport systems in the yeast Pichia ohmeri. AB - Glucose-repressed cells of the yeast Pichia ohmeri IGC 2879 transported glucose by facilitated diffusion. Derepression led to the formation of a glucose/proton symport and the simultaneous reduction of the facilitated diffusion capacity by about 70%. Cycloheximide prevented this interconversion indicating its dependence on de novo protein synthesis (proteosynthetic interconversion). In buffer with 2% glucose the glucose/proton symport suffered irreversible inactivation while the facilitated diffusion system was simultaneously restored. This reverse interconversion process did not require de novo protein synthesis as indicated by its lack of sensitivity to cycloheximide (degradative interconversion). Thus the glucose/proton symport system appeared to consist of about 70% of the facilitated diffusion proteins turned silent through association with additional protein(s) the latter being sensitive to glucose-induced repression and glucose-induced inactivation. PMID- 3036222 TI - The effects of cycloheximide on Na+/H+ antiporter activity in cultured opossum kidney cells. AB - These studies examined the effects of cycloheximide on the Na+/H+ antiporter in cultured opossum kidney cells. The effects of cycloheximide on antiporter activity depended on the basal level of activity. These data suggest that the Na+/H+ antiporter may be regulated by several processes which are sensitive to protein synthesis inhibition. PMID- 3036223 TI - Optimal posttranslational translocation of the precursor of PhoE protein across Escherichia coli membrane vesicles requires both ATP and the protonmotive force. AB - In order to reach their final destination, periplasmic and outer membrane proteins have to pass the cytoplasmic membrane of Escherichia coli cells. To study the transport of PhoE protein, we developed an in vitro transcription translation and translocation system. In this in vitro system, the protein is synthesized as a larger precursor, which can be processed by purified leader peptidase. The precursor can be translocated into inverted inner membrane vesicles as judged by the protection against externally added protease. Only part of the translocated protein is in the processed mature form. Translocation can occur posttranslationally and requires both ATP and the protonmotive force for an optimal process. Upon incubation of vesicles with mature PhoE protein or precursor PhoE in the absence of ATP, the proteins are bound to the vesicles, but they are not translocated, since they are still sensitive to externally added protease. PMID- 3036225 TI - A new paramagnetic analogue of cholesterol as a tool for studying molecular interactions of genuine cholesterol. AB - The synthesis of a new paramagnetic (nitroxide) analogue of cholesterol is described. This compound (called CNO) contains a doxyl group in the lateral chain at position 25. Our results show that CNO retains three molecular interactions which characterize authentic cholesterol: It assumes an orientation perpendicular to the phospholipid bilayer with the doxyl group buried in the membrane core, as seen by ESR spectroscopy. It widens the transition temperature of dimyristoylphosphatidylcholine, to the same extent as cholesterol, as measured by Raman and ESR spectroscopies. It interacts with polyene antibiotics, such as amphotericin B and filipin, in the same manner as its model. This was proved on the one hand by the change in fluorescence of self quenched vesicle-entrapped calcein, after dilution in the external medium, provoked by filipin, and on the other hand by fluorescence quenching provoked by cobalt ions entering the vesicles under the influence of amphotericin B. We concluded that CNO, although it has a side chain different from genuine cholesterol, can help to solve many physiologically meaningful questions related to the distribution and movement rate of cholesterol itself. PMID- 3036224 TI - Vanadate inhibition of ATP-dependent H+ transport in membrane vesicles from turtle bladder epithelial cells. AB - The ATP-dependent proton transport into vesicles of a mixed membrane fraction obtained from turtle bladder epithelial cells consists of at least two kinetically defined moieties: one, which is maximally inhibited by 25% with nanomolar levels of vanadate, but not inhibited at all with equimolar levels of N ethylmaleimide, and another, which is maximally inhibited by 70% with micromolar levels of N-ethylmaleimide and by 25% with equimolar levels of vanadate. In contrast to the transport function, the associated enzymatic function (the ouabain-resistant ATPase activity) in these membranes, not inhibited by nanomolar levels of vanadate or N-ethylmaleimide, is maximally inhibited by 40% with micromolar levels of vanadate and by 13% with equimolar levels of N ethylmaleimide. Independent of these kinetic differences between the enzyme and the transport functions, membranes containing the N-ethylmaleimide-sensitive proton transport function are electrophoretically separable from those containing the vanadate-sensitive transport function. For example, the kinetically defined, vanadate-sensitive proton transport function is recovered exclusively and kinetically identified in one of four electrophoretic membrane fractions, EF-II; while the N-ethylmaleimide-sensitive function is recovered in EF-III as well as in EF-II. Membranes of EF-IV, maximally enriched in ouabain-resistant ATPase activity, possess no proton transport function at all, even in the absence of N ethylmaleimide or vanadate. Additional data under in vivo as well as under in vitro conditions are required to prove that the vanadate-sensitive proton transport in these vesicles is an in vitro manifestation of the mechanism responsible for generating the vanadate-sensitive luminal acidification process under in vivo conditions in the intact turtle bladder. PMID- 3036226 TI - Specific interaction of arabinose residue in ginsenoside with egg phosphatidylcholine vesicles. AB - The interaction of the specific sugar residue in ginsenosides with egg phosphatidylcholine vesicles was investigated by ESR spectrometry using phosphatidic acid spin-labeled at the polar head groups. Ginsenoside-Rc, which has an alpha-L-arabinofuranose residue and agglutinability toward egg yolk phosphatidylcholine vesicles (Fukuda, K. et al. (1985) Biochim. Biophys. Acta 820, 199-206), caused the restriction of the segmental motion of spin-labeled phosphatidic acid in egg phosphatidylcholine vesicles, indicating that the saponin interacted with the polar head groups of vesicles. Other ginsenosides Rb2, Rb1, Rd and p-nitrophenyl glycoside derivatives which have less or no agglutinability were also investigated in the same manner. Only ginsenoside-Rb2 and p-nitrophenyl alpha-L-arabinofuranoside which have the specific sugar residue (arabinose) showed a strong interaction with the polar head groups of vesicles. To gain an insight into the mechanism of agglutination by ginsenoside-Rc, the interaction with the fatty acyl groups was also studied by using phosphatidylcholine spin-labeled at the fatty acyl groups. Ginsenoside-Rc increased the order parameter of the spin-labeled phosphatidylcholine, indicating that the saponin was inserted into lipid bilayers. In other saponins investigated, only ginsenoside-Rb2 interacted with the fatty acyl part of vesicles. The process of expression of agglutination by ginsenoside-Rc was discussed on the basis of the ESR studies. PMID- 3036227 TI - Incorporation of highly purified melittin into phosphatidylcholine bilayer vesicles. AB - Melittin free of phospholipase A2 was prepared. In the absence of salt this highly pure protein starts to aggregate in solution at a protein concentration of Cp greater than 10(-3) M. In high salt solution (2 M) aggregation starts at Cp greater than 10(-6) M. This was determined from the blue shift of the intrinsic fluorescence of the protein. Reinvestigation of the quenching behaviour clearly shows that self-aggregation cannot be deduced from quenching experiments using nitrate or 2,2,6,6-tetramethylpiperidine-1-oxyl as quencher. The incorporation of melittin into phosphatidylcholine bilayer vesicles was studied by fluorescence quenching and by energy-transfer experiments using 2- and 6-anthroyloxypalmitic acid as acceptor and peptide tryptophan as donor. Incorporation of melittin into small unilamellar vesicles was found to be reduced below the lipid phase transition temperature, Tt, whereas it incorporates and distributes more randomly above Tt. Cooling the temperature below Tt after incubation at T greater than Tt leads to a deeper incorporation of the peptide into the lipid bilayer due to electrostatic interaction between the lipid phosphate groups and the positively charged amino acids. This stabilizing effect is lost above Tt and melittin is extruded to the polar phase. Quenching experiments support this finding. EPR measurements clearly demonstrate that even in the presence of high amounts of melittin up to 10 mol% with respect to the lipid broadening of the phase transition curves was only observed with fatty acid spin labels, where the doxyl group is localized near the bilayer surface. The order degree of the inner part of the bilayer remains almost unchanged even in the presence of high melittin content. PMID- 3036228 TI - Glycolipid function. PMID- 3036229 TI - Vesicular stomatitis virus membrane proteins and their interactions with lipid bilayers. PMID- 3036231 TI - Nucleotide sequence of two 3'-termini of rRNA in the sea urchin Lytechinus variegatus. AB - The 3'-terminus of 26 S rRNA from the sea urchin Lytechinus variegatus has been determined by oligonucleotide fingerprinting, S1 nuclease mapping and terminal nucleotide analysis. There are two species of 26 S rRNA of approximately equal abundance, one 19 nucleotides longer than than other. PMID- 3036230 TI - Mutants of murine leukemia viruses and retroviral replication. AB - The analysis of retroviral mutants has played a critical role in the development of our understanding of the complex viral life cycle. The most fundamental result of that analysis has been the definition of the replication functions encoded by the viruses. From a biochemical examination of a particular step in the life cycle it is difficult to determine, for example, whether that step is catalyzed by a viral or a host enzyme; but the isolation of a viral mutant defective in that step can firmly establish that a viral function is involved. In this way many facts about the viruses have been established. We know that reverse transcriptase is encoded by the virus; that RNAase H and DNA polymerase activities reside on the same gene product; that processing of many precursor proteins is mediated by a viral proteinase; and that establishment of the integrated provirus requires a viral protein. The list of functions mediated by viral enzymes has largely been defined by the mutants isolated and studied in various laboratories. The second significant result of the studies of viral mutants has been the assignation of the replication functions to particular viral genes, and then more specifically to particular domains of these genes. Mutants and viral variants have been essential in the determination, for example, that the gag protein is the critical gene product for the assembly of a virion particle; that the env protein is the determinant of species specificity of infection; or that the LTR is a major determinant of tissue tropism and leukemogenicity. The subdivisions of functions within a given gene have similarly hinged on mutants. Genetic mapping was needed to establish that P30 is the most important region for assembly; that the proteinase and integrase functions reside, respectively, in the 5' and 3' portions of the pol gene; and that the glycosylated gag protein is dispensable for replication. A third important area of knowledge has depended heavily on viral mutants: the determination of host functions and proteins that interact with viral proteins. Variant viruses with altered or restricted host ranges serve to define differences between pairs of different host cells, and the mapping of the viral mutations serves to define the viral protein important in that interaction with the host. These studies are only in their infancy, but it is clear that substantial efforts will be made to further analyze these host functions.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3036232 TI - Modulation of testicular galactolipid sulfotransferase activity in vitro by ATP. AB - Testicular galactolipid sulfotransferase activity is an early marker of differentiation during mammalian spermatogenesis. The enzyme will catalyze the sulfation of galactosylglycerol in the 3' position of the galactose moiety at 37 degrees C in vitro. However, sulfotransferase activity was found to be completely lost on preincubation of the solubilized enzyme preparation at 37 degrees C. This loss of activity was completely prevented by inclusion of ATP and Triton in the preincubation step. This protective effect was synergistic, pH dependent and correlated with an inhibition of endogenous phosphatase activity. These results are interpreted to suggest that the galactolipid sulfotransferase may be regulated by a phosphorylation mechanism. PMID- 3036233 TI - The effect of complex formation upon the reduction rates of cytochrome c and cytochrome c peroxidase compound II. AB - The effect of complex formation between ferricytochrome c and cytochrome c peroxidase (Ferrocytochrome-c:hydrogen peroxide oxidoreductase, EC 1.11.1.5) on the reduction of cytochrome c by N,N,N',N'-tetramethyl-p-phenylenediamine (TMPD), reduced N-methylphenazonium methosulfate (PMSH), and ascorbate has been determined at low ionic strength (pH 7) and 25 degrees C. Complex formation with the peroxidase enhances the rate of ferricytochrome c reduction by the neutral reductants TMPD and PMSH. Under all experimental conditions investigated, complex formation with cytochrome c peroxidase inhibits the ascorbate reduction of ferricytochrome c. This inhibition is due to the unfavorable electrostatic interactions between the ascorbate dianion and the negatively charged cytochrome c-cytochrome c peroxidase complex. Corrections for the electrostatic term by extrapolating the data to infinite ionic strength suggest that ascorbate can reduce cytochrome c peroxidase-bound cytochrome c faster than free cytochrome c. Reduction of cytochrome c peroxidase Compound II by dicyanobis(1,10 phenanthroline)iron(II) (Fe(phen)2(CN)2) is essentially unaffected by complex formation between the enzyme and ferricytochrome c at low ionic strength (pH 6) and 25 degrees C. However, reduction of Compound II by the negatively changed tetracyano-(1,10-phenanthroline)iron(II) (Fe(phen)(CN)4) is enhanced in the presence of ferricytochrome c. This enhancement is due to the more favorable electrostatic interactions between the reductant and cytochrome c-cytochrome c peroxidase Compound II complex then for Compound II itself. These studies indicate that complex formation between cytochrome c and cytochrome c peroxidase does not sterically block the electron-transfer pathways from these small nonphysiological reductants to the hemes in these two proteins. PMID- 3036234 TI - Phosphotransferase activity of human alkaline phosphatases and the role of enzyme Zn2+. AB - Purified isoenzymes of human alkaline phosphatase from placenta, intestine and liver were investigated as catalysts for phosphotransferase activity, using the phosphoacceptors Tris, 2-amino-2-methyl-1-propanol, 2-amino-2-methyl-1,3 propanediol, diethanolamine, 2-(ethylamino)ethanol, ethanolamine, and N-methyl-D glucamine. All of the compounds supported phosphotransferase catalysis, conforming to saturation kinetics. There was little difference among the isoenzymes with respect to Km values of the acceptors, but the liver form was the most efficient (highest Vmax/Km) in forming phosphoacceptors; it was also the most efficient (highest Vamax/Ka) when the phosphoacceptors were considered as activators. At Vmax the isoenzymes differed little in their support of phosphotransferase activity relative to phosphohydrolysis, although the intestinal enzyme tended to be the poorest. The two best acceptors were diethanolamine, providing the highest phosphotransferase velocity, and 2 (ethylamino)ethanol, having the lowest Km. The phosphoaceptors that bound Zn2+ tightly did not function well in the phosphotransferase reaction, and vice versa. However, temporal assessment of the phosphohydrolytic and phosphotransferase activities during removal of Zn2+ from the enzyme with 1,10-phenanthroline revealed no evidence of a special role for Zn2+ in the latter activity. PMID- 3036235 TI - 31P-NMR study of the orotate phosphoribosyltransferase equilibrium with thiopyrophosphate as substrate. AB - 31P-nuclear magnetic resonance spectroscopy was used to directly determine the equilibrium of the reaction catalysed by yeast orotate phosphoribosyltransferase, using orotidine monophosphate and inorganic pyrophosphate as substrates. A Keq value of 0.71 was determined, in good agreement with that of 0.49 calculated by Victor, Greenberg and Sloan (J. Biol. Chem. 254 (1979) 2647-2655), from kinetic data. Substitution of thiopyrophosphate as the substrate shifted the position of the equilibrium 55-fold, to yield a Keq value of 39. Only the beta S analogue of 5-phosphoribosyl 1-diphosphate appeared to be synthesized in this reaction. PMID- 3036236 TI - Phospholipase C from human sperm specific for phosphoinositides. AB - Human sperm lysates were incubated in the presence of 1-[14C]stearoyl-2-acyl-sn glycero-3-phosphocholine, 1-[14C]stearoyl-2-acyl-sn-glycero-3-phosphoethanolamine or 1-[14C]stearoyl-2-acyl-sn-glycero-3-phosphoinositol. Only the latter substrate was hydrolyzed to a significant extent, with a concomitant formation of 1 [14C]stearoyl-2-acyl-sn-glycerol. Furthermore, incubation of phosphatidyl[3H]inositol under the same conditions was accompanied by the formation, in roughly equal amounts, of [3H]inositol 1-phosphate and [3H]inositol 1:2-cyclic monophosphate. Finally [32P]phosphatidylinositol 4-phosphate and [32P]phosphatidylinositol 4,5-bisphosphate were degraded into [32P]inositol 1,4 bisphosphate and [32P]inositol 1,4,5-trisphosphate, respectively. The phosphoinositide-specific phospholipase C was activated by calcium (optimal concentration 5-10 mM) and inhibited by EGTA, although endogenous calcium supported a half-maximal activity. The enzyme displayed an optimal pH of 6.0 and an apparent Km of 0.08 mM. Its specific activity was around 10 nmol/min per mg protein, which is approximately the same as that found in human blood platelets. Subcellular fractionation revealed that 55% of the enzyme was solubilized under conditions where 80% of acrosin appeared in the supernatants. The majority of the particulate phospholipase C activity (37% of total) was found in the 1000 X g pellet, which contained only 8% of total acrosin activity. Further fractionation of spermatozoa into heads and tails indicated no specific enrichment of phospholipase C activity in any of these two fractions. However, owing to a 4 fold higher protein content in the head compared to the tail fraction, it is concluded that about 80% of particulate phospholipase C activity is located in sperm head. The physiological significance of this enzyme is discussed in relation to a possible role in acrosome reaction and (or) in egg fertilization. PMID- 3036237 TI - The effect of variations in lipid composition of high-density lipoprotein on its interaction with receptors on human fibroblasts. AB - To test whether the altered lipid composition of high-density lipoprotein (HDL) particles influences their ability to interact with the HDL receptor on cultured fibroblasts, HDL3 isolated from normal and diabetic donors with different degrees of hypertriglyceridemia was subjected to binding competition, cholesterol efflux, and net cholesterol transport assays. When HDL3 particles from different subjects were incubated with cholesterol-loaded fibroblasts, the initial rates of cholesterol efflux from cells to HDL3 particles appeared to be an exclusive function of the relative ability of HDL3 to interact with the HDL receptor. Variation in lipid composition of HDL3 particles did not appear to have any significant influence on either the receptor-binding or the efflux-promoting abilities of HDL3. When the movement of cholesterol between cells and HDL3 particles was allowed to approach equilibrium, the lipid composition of HDL3 became an important factor in determining the net amount of cholesterol removed from cells, with cholesterol-deficient triacylglycerol-rich HDL3 particles having the best capacity to promote net transport of cholesterol from cells. These results suggest that the ability of HDL to bind to its cell-surface receptor, rather than variations in the lipid composition of the HDL particle, is the major determinant of cholesterol efflux from cells to HDL particles. However, the lipid composition of HDL as well as its receptor-binding activity determine the net amount of cholesterol transported from cells over long-term incubation. PMID- 3036238 TI - The metabolism of lyso-platelet-activating factor (1-O-alkyl-2-lyso-sn-glycero-3 phosphocholine) by a calcium-dependent lysophospholipase D in rabbit kidney medulla. AB - A Ca2+-dependent lysophospholipase D activity in microsomal preparations from the rabbit kidney medulla hydrolyzes the choline moiety from 1-O-[9,10-3H]hexadecyl-2 lyso-sn-glycero-3-phosphocholine (lyso-PAF) to form 1-O-[9,10-3H]hexadecyl-2-lyso sn-glycero-3-P; the latter is subsequently dephosphorylated by a phosphohydrolase to 1-O-[9,10-3H]hexadecyl-sn-glycerol. Sodium vanadate, which is known to inhibit phosphohydrolases, reduces the proportion of hexadecylglycerol and increases the formation of hexadecyl-lysoglycerophosphate. Essentially no hydrolysis occurs when the sn-2 position of the hexadecyllysoGPC substrate contains an acyl moiety. The lysophospholipase D in rabbit kidney is of microsomal origin and has a broad pH optimum between 8.0 and 8.8, with the activity decreasing sharply from pH 7.6 to 7.2. Wykle et al. (Biochim. Biophys. Acta 619 (1980) 58-67) have previously demonstrated the existence of a microsomal lysophospholipase D (specific for ether lipid substrates) in rat tissues that requires Mg2+ and exhibits a pH optimum of 7.2; high activities of the Mg2+-dependent lysophospholipase D were found in liver and brain, but not in kidney. In contrast to the Mg2+-dependent lysophospholipase D in rat tissues, the renal enzyme from rabbits requires Ca2+ (5 mM), whereas Mg2+ (5 mM) exhibits little stimulatory action. Under optimal assay conditions (0.1 M Tris-HCl (pH 8.4)/5 mM CaCl2), lysophospholipase D in the rabbit kidney medulla has an activity of 2.7 nmol/min per mg protein compared to 0.9 nmol/min per mg protein for the lysophospholipase D in the rat kidney medulla (0.1 M Tris-HCl (pH 7.2)/5 mM MgCl2). The Ca2+-dependent lysophospholipase D is highest in the liver and kidney medulla from rabbits, but is very low in rat tissues; similar activities were found in male and female rabbits. Our data indicate that the divalent metal ion requirements for expression of maximum lysophospholipase D activities can differ markedly among animal species and also suggest the microsomal Ca2+-dependent lysophospholipase D is an important catabolic route for lyso-PAF metabolism in rabbit renomedullary tissue. PMID- 3036239 TI - Tyrosinase-catalyzed oxidation of dopa and related catechol(amine)s: a kinetic electron spin resonance investigation using spin-stabilization and spin label oximetry. AB - The oxidation of four catechol(amine)s by tyrosinase has been studied by electron spin resonance and optical methods. Rates of oxygen consumption and of dopaquinone and dopachrome formation during the oxidation of dopa have been measured, and compared with rates of dopasemiquinone production measured using spin-stabilization procedures. In the presence of spin-stabilizing metal ions, production of semiquinone is approximately quantitative. Time-dependent ESR spectra obtained from dopa and dopamine show a slow regeneration of semiquinone, suggesting that a semiquinone precursor is slowly reformed. In contrast, time dependent spectra for 4-methylcatechol and N-acetyldopamine show decay of the primary semiquinone together with buildup of a secondary semiquinone apparently derived from the corresponding 6-hydroxy-catechol(amine). Thus, catecholamines that give rise to a cyclizable quinone show a pattern of behavior that differs from those that produce a non-cyclizable quinone. These results are discussed in terms of their possible significance to melanogenesis and the toxicity of catechol(amine)s, which has been attributed to production of semiquinones and/or other oxygen radicals. PMID- 3036240 TI - Photogeneration of free radicals from eumelanogenic intermediates and metabolites. AB - We have recently demonstrated that cysteinyldopas, pheomelanogenic precursors and excreted eumelanogenic metabolites, are photolabile and initiate DNA damage in vitro. In this study we have extended our photochemical investigations to eumelanogenic indole intermediates and metabolites. Continuous-wave photolysis of 5,6-dihydroxyindole (DHI), 5,6-dihydroxyindole-2-carboxylic acid (DHICA), or its 5-methoxylated metabolite (HMICA) with biologically relevant ultraviolet radiation (i.e., wavelengths greater than 300 nm) resulted in rapid destruction of starting material. Using ESR spin-trapping techniques we observed the initial production of free radical species; prolonged photolysis resulted in the formation of polymeric photoproducts. Radicals were trapped by the nitrone spin trap 5,5-dimethyl-1-pyrroline N-oxide (DMPO) and characterized by their ESR spectra as hydrated electrons and hydrogen atoms. Experiments further demonstrated that while DHI photoionizes, the two indole-2-carboxylic acid derivatives do not ionize appreciably upon irradiation, rather homolysis of X-H bonds appears to be a significant photochemical pathway. The potential photobiological significance of melanogenic indole intermediates is discussed. PMID- 3036241 TI - Gluconate metabolism in germinated spores of Bacillus megaterium QM B1551: primary roles of gluconokinase and the pentose cycle. AB - The metabolic pathway of gluconate, a major product of glucose metabolism during spore germination, was investigated in Bacillus megaterium QM B1551. Compared to the parent, mutant spores lacking gluconokinase could not metabolize gluconate, whereas the revertant simultaneously restored the enzyme activity and the ability to metabolize it, indicating that gluconokinase was solely responsible for the onset of gluconate metabolism. To identify a further metabolic route for gluconate, we determined 14C yields in acetate and CO2 formed from [14C]gluconate, and found that experimental ratios of 14CO2/[14C]acetate obtained from [2-14C]gluconate and [3,4-14C]gluconate were not compatible with the ratios predicted from the Entner-Doudoroff pathway. In contrast, when CO2 release caused by recycling (approx. 30%) was corrected, the ratios almost agreed with those from the pentose cycle. Comparison of specific radioactivities in acetate also supported the conclusion that gluconate was metabolized via the pentose cycle, subsequently metabolized via the Embden-Meyerhof pathway, and finally degraded to acetate and CO2 without a contribution by the Krebs cycle. PMID- 3036242 TI - The activating effects of bicarbonate on sperm motility and respiration at ejaculation. AB - Mature porcine sperm preserved in the cauda epididymis are quiescent. At ejaculation, they are mixed with the seminal vesicle fluid containing HCO3- and are rapidly activated. The role of HCO3- on the sperm activation process at ejaculation was studied in vitro. HCO3- quickly increased the motility, respiration rate and cAMP content of the porcine epididymal sperm. The extent of activation was proportional to the pCO2 in the medium. The activating effect of HCO3- on the motility was observed even in the absence of fructose as well as in the presence of KCN. 8-Bromoadenosine 3',5'-cyclic monophosphate and theophylline showed similar activating effects to that of HCO3-. However, HCO3(-)-free seminal plasma, Ca2+, amino acids, intermediates of the Krebs cycle, substrates of respiration and increases in the intracellular pH, extracellular pH or ionic strength of the medium had no effect. Fructose sustained the active state of the sperm and gradually increased both the motility and respiration rate when the dose of HCO3- was low. The anion channel blocker enhanced the activating effect of HCO3-. These results suggest that, upon ejaculation, HCO3- is a unique activator in vivo which makes the quiescent sperm motile via the HCO3(-) adenylate cyclase-cAMP system, to which an endogenous HCO3- derived from metabolic CO2 may be related. PMID- 3036243 TI - Similarities of physico-chimical and antigenic properties of neurocupreins and extremely acidic copper proteins from chromaffin granules. AB - Extremely acidic copper-containing proteins, neurocupreins, were isolated from brains of various mammals (bovine, rabbit, pig and sheep). Neurocupreins from all these sources were found to have similar physico-chemical and antigenic properties. Using the immunological approach, it was shown that neurocuprein is located only in brain cytosol and synaptosomal fractions. Extremely acidic copper containing proteins were also isolated from soluble and membranous fractions of chromaffin granules from bovine adrenal medulla. The soluble form of the protein from the granules has practically the same physico-chemical and antigenic properties as neurocupreins. The copper protein isolated from membranes of granules has slightly higher molecular weight and somewhat different amino acid composition, although their EPR spectra are identical. However, both copper proteins from chromaffin granules are immunoprecipitated with antibodies to neurocuprein. It is suggested that the membranous form differs from the soluble one in possessing a peptide which prolongs the protein chain without changes in its antigenic properties. PMID- 3036244 TI - Occurrence of vanadium ion in serum albumins. AB - Vanadium was shown to be a contaminant in Sigma-grade serum albumins, fraction V. The levels of vanadium detected by neutron activation analysis were: bovine 16.7, ovine 10.0, rabbit 9.1, porcine 3.9, and human 0.19 micrograms/g protein. According to the ESR spectra, the vanadate form (+5 oxidation state) was strongly suggested as a chemical form present in albumins. Dialysis against a chelating agent was quite effective for removal of the metal ion. PMID- 3036245 TI - Taurine inhibits octopamine-stimulated cAMP production in cockroach haemocytes. AB - Taurine and the taurine analogue guanidinoethanesulfonic acid interact with octopamine receptors of cockroach hemocytes to decrease octopamine-stimulated cAMP production. Dopamine-, synephrine- and tyramine-stimulated cAMP production in the haemocytes are also inhibited by taurine. PMID- 3036247 TI - Bryostatins mimic the effects of phorbol esters in intact human platelets. AB - Bryostatin-7 induces aggregation of human platelets and the phosphorylation of specific platelet proteins. Both the rate and extent of aggregation are similar to that induced by the tumor promoter phorbol ester 12-myristate 13-acetate (PMA); however, the rate of response is markedly reduced compared to that induced by thrombin. The addition of bryostatin-7 to 32P-labeled platelets results in a time-dependent incorporation of 32P into proteins of 20, 47 and 250 kDa; proteins of similar molecular mass are phosphorylated in response to the addition of thrombin or PMA. The time courses and dose responses of the phosphorylations induced by bryostatin-7 are similar to those found with PMA. In addition, bryostatin-7 increases the level of 32P incorporation into platelet polyphosphoinositides, which also occurs in response to PMA. These results suggest that, in intact human platelets, bryostatin-7 mimics the phorbol ester tumor promoter by directly activating protein kinase C. PMID- 3036246 TI - Transferrin and ferritin endocytosis and recycling in guinea-pig reticulocytes. AB - Transferrin and ferritin endocytosis and exocytosis by guinea-pig reticulocytes were studied using incubation with pronase at 4 degrees C to distinguish internalized and membrane-bound protein. Internalization of both transferrin and ferritin occurred in a time- and temperature-dependent fashion. Transferrin endocytosis was more rapid than that of ferritin. Transferrin binding to receptors was not altered, but transferrin endocytosis was decreased in the presence of ferritin. Iron accumulation from transferrin was inhibited by ferritin to a greater extent than could be accounted for by the decreased rate of endocytosis. In pulse-chase experiments, almost all of the transferrin was released intact from reticulocytes, but only about 50% of the total internalized ferritin was released, of which 85% was intact. The endocytosis of transferrin by rabbit reticulocytes was 2- to 2.5-times faster than guinea-pig reticulocytes. These data suggest that ferritin and transferrin are internalized by receptor mediated endocytosis, possibly involving the same coated pits and vesicles, but that the proteins are recycled only partly in common. PMID- 3036248 TI - Glucagon receptor binding, dissociation and degradation in rat liver plasma membranes studied by a microperifusion method. AB - The association process of glucagon receptor binding in purified rat liver plasma membranes and prolonged incubation of the hormone-receptor complex at 30 degrees C did not result in degradation of bound labelled glucagon. In contrast, up to 95% of the non-membrane-bound labelled glucagon was degraded. The rate of spontaneous dissociation of the glucagon-receptor complex was slow, and amounted to about 0.1% per min of that bound. GTP greatly enhanced the rate of dissociation. Half the maximal dissociation of the complex was effected by 10(-5) mol/l of GTP under equilibrium binding conditions. At maximally effective concentrations of GTP, 80% of the glucagon-receptor complex was dissociated within 2 min. A microperifusion system for the perifusion of isolated plasma membranes was devised and used for the separation of labelled glucagon from the plasma membranes subsequent to a GTP-induced dissociation of the hormone-receptor complex. Rebinding of the dissociated peptide to fresh membranes showed that maximum binding ability was retained. The glucagon molecule was protected against degradation while bound to the receptor, indicating that the glucagon effector system is completely separate from the inactivating system(s) in isolated plasma membranes. Thus, the hormonal effect of glucagon could be exerted through the sequential interaction of each glucagon molecule with several receptors. PMID- 3036249 TI - Activation of phospholipase C in platelets by platelet activating factor and thrombin causes hydrolysis of a common pool of phosphatidylinositol 4,5 bisphosphate. AB - Despite their physicochemical and mechanistic differences platelet activating factor (or acetylglycerylether phosphorylcholine; AGEPC) and thrombin, both platelet stimulatory agents, induce phosphoinositide turnover in platelets. We therefore investigated the stimulation of the phosphoinositide phosphodiesterase by these agents and questioned whether they evoked hydrolysis of the same or different pools of phosphoinositides. [3H]Inositol-labelled rabbit platelets were challenged with thrombin and/or AGEPC under a variety of protocols, and the phospholipase C mediated production of radioactive inositol monophosphate (IP); inositol bisphosphate (IP2) and inositol trisphosphate (IP3) was used as the parameter. AGEPC (1 X 10(-9) M) caused a transient maximum (5 to 6-fold) increase in [3H]IP3 at 5 s followed by a decrease. Thrombin (2 U/ml) elicited an increase in [3H]IP3 at a much slower rate than AGEPC; 2 fold at 5 s, 5 fold at 30 s and a maximum 6 to 8-fold at 2-5 min. Compared to AGEPC, thrombin stimulated generation of [3H]IP2 and [3H]IP were severalfold higher. When thrombin and AGEPC were added together to platelets there was no evidence for an additive increase in inositol polyphosphate levels except at earlier time points where increases were submaximal. When AGEPC was added at various time intervals after thrombin pretreatment, no additional increases in [3H]IP3 were observed over that maximally seen with thrombin or AGEPC alone. In another set of experiments, submaximal increases (about 1/4 and 1/2 of maximum) in [3H]IP3 were achieved by using selected concentrations of thrombin (0.1 U and 0.3 U, respectively) and then AGEPC (1 X 10(-9) M) was added for 5 s. Once again the increase in [3H]IP3 was close to the maximal level seen with thrombin or AGEPC individually. It is concluded that thrombin and AGEPC differentially activated phosphoinositide phosphodiesterase (phospholipase C) in rabbit platelets and that the stimulation of the phospholipase C by these two stimuli causes IP3 production via hydrolysis of a common pool of phosphatidylinositol 4,5-bisphosphate. PMID- 3036250 TI - Procalpain is activated on the plasma membrane and the calpain acts on the membrane. AB - The mechanism of activation of human erythrocyte calpain was investigated using the immunoblotting technique with anticalpain monoclonal antibody. The purified calpain underwent a Ca2+-induced fragmentation of the 80 kDa subunit to 76 kDa and 36 kDa fragments. The behavior of the 76 kDa fragment in electrophoresis corresponded to the proteinase activity of calpain, whereas the behavior of the 80 kDa subunit and the 36 kDa fragment did not. When inside-out membrane vesicles were added to the reaction mixture of calpain and Ca2+ and the vesicles were separated from the supernatant solution by centrifugation, the 80 kDa subunit and 76 kDa fragment were found in the vesicle fraction. No other fragments were found in this fraction. On the other hand, the 80 kDa subunit and 36 kDa fragment were found in the supernatant fraction. When right-side-out membrane vesicles were added to the reaction mixture and the vesicles were separated from the supernatant fraction, no fragment was found in the vesicle fraction, while only the 36 kDa fragment was found in the supernatant fraction. These results indicate that the 80 kDa subunit of procalpain was bound in a Ca2+-dependent manner to the cytosolic surface of the plasma membrane and then underwent fragmentation to produce the 76 kDa fragment (active form) and that it expressed its proteinase activity at the surface of the membrane. PMID- 3036251 TI - Receptor binding of selectively labeled (Tyr-10) and (Tyr-13)-mono-125I-glucagons and competition by homologous 127I-labeled isomers. AB - Two monoiodinated derivatives of glucagon were prepared by lactoperoxidase catalyzed iodination followed by separation on reverse-phase high-performance liquid chromatography. The purified (Tyr-10) and (Tyr-13)-mono-125I-labeled glucagon isomers were characterized and studied with respect to their binding to the receptors of isolated intact rat hepatocytes. The extent of steady-state binding to cellular receptor sites differed for the two labeled glucagon tracers at 37 degrees C as well as at 15 degrees C with (Tyr-10)-mono-125I-glucagon displaying higher receptor binding. The apparent equilibrium constants, Kd,app at 37 degrees C are 3.6 +/- 0.4 nM (mean +/- S.E. of three independent experiments) for the tyrosine-13-labeled tracer and 5.9 +/- 0.6 nM for the tyrosine-10-labeled glucagon with native glucagon as competitor. Since the observed Kd in the competition assay is a function of the true Kd values of the monoiodinated radioactive glucagon isomers and native glucagon, the dissociation constants were also measured with chemically identical tracer and competitor. Under these conditions, we obtained Kd values of 1.3 +/- 0.2 nM for the tyrosine-10-labeled analog and 2.0 +/- 0.2 nM for the tyrosine-13-labeled glucagon isomers confirming the higher receptor binding affinity of (Try-10)-mono-125I-glucagon. All competition curves fit the mathematical expression for a model of non-cooperative binding to a single class of receptors. PMID- 3036253 TI - Do "d-blob" and "l-blob" hypercolumns tessellate the monkey visual cortex? AB - The orientation selective neurons in the monkey striate cortex seem to be organized in pairs of mirror symmetrical hypercolumnar patches, each of which centered by a "cytochrome oxidase blob." A simple scheme derived from recent data of Blasdel and Salama reconciles earlier models assuming either linear or circular representation of the preferred direction of edges in the visual field. PMID- 3036252 TI - Analysis of the in vivo phosphorylation state of protein phosphatase inhibitor-2 from rabbit skeletal muscle by fast-atom bombardment mass spectrometry. AB - A new procedure has been developed for identifying phosphoserine residues in proteins, and is used to analyse the in vivo phosphorylation state of inhibitor 2. The method employs reverse-phase liquid chromatography to resolve phosphorylated and dephosphorylated forms of peptides and fast-atom bombardment mass spectrometry (FABMS) to identify phosphorylated derivatives. The positions of phosphorylation sites within peptides are located by gas-phase sequencer analysis after conversion of phosphoserine residues to S-ethylcysteine. The phosphorylation sites on inhibitor-2 were identified as serines-86, -120 and 121, the three residues phosphorylated in vitro by casein kinase-II. Serine-86 was phosphorylated to 0.7 mol/mol and serines-120 and -121 each to 0.3 mol/mol. These values were not altered significantly by intravenous injection of adrenalin or insulin. No phosphate was present in the region comprising residues 1-49, even after injection of adrenalin, demonstrating that inhibitor-2 is not a substrate for cyclic AMP-dependent protein kinase in vivo. The absence of phosphotyrosine also indicated that inhibitor-2 is not a physiological substrate for the insulin receptor. Surprisingly, no phosphate was present at threonine-72, the residue phosphorylated in vitro by glycogen synthase kinase-3, after injection of either propranolol, adrenalin or insulin. The implications of this finding for the in vivo activation of protein phosphatase 1I (the 1:1 complex between inhibitor-2 and the catalytic subunit of protein phosphatase-1) are discussed. FABMS analysis of inhibitor-2 confirmed the accuracy of the primary structure reported previously, and showed that the only post-translational modifications were an N acetyl moiety and the three phosphoserine residues. FABMS also demonstrated the presence of an additional serine residue at the C-terminus, and showed that 50% of isolated inhibitor-2 molecules lack the C-terminal Ser-Ser dipeptide. PMID- 3036254 TI - On periodic responses generated by a degenerate analog neuron model with periodic inputs. AB - Periodic responses to periodically varying stimulating pulse sequences are mathematically described for a degenerate analog neuron model. The model used was derived by Yoshizawa et al. (1982) in their investigation of the state transition of an electronic model using a tunnel diode in a degenerate case. Periodic responses to constant pulse sequences for the model were described by them. In this paper, it is shown that periodic responses to periodically varying pulse sequences for the model are identical with those which the author previously described for a discrete neuron model. PMID- 3036255 TI - On the identification of neural responses. AB - The most common form of measuring electrical responses of nerve cells is the recording of a given cell's "spike train" profile to the parameters of a given input signal. In this paper we consider the conditions under which it is possible to relate such response measures to the properties of the cell's underlying activity characteristics, the neural network, and the input signal. PMID- 3036256 TI - [Oxidation by nitrite of azurin and cytochrome c-551 from Pseudomonas aeruginosa]. AB - The nitrite oxidizes reduced azurin and cytochrome c-551 from Pseudomonas aeruginosa. The effects of pH, ionic strength and concentrations of nitrite, EDTA and the protein on the oxidation were investigated. The results obtained indicate that nitrite interacts not only with the terminal electron carrier of the nitrite reducing chain (nitrite reductase, cytochrome cd1) but also with the intermediate electron carrier components of the chain (azurin and cytochrome c-551). PMID- 3036257 TI - [Stimulation of transcription in mouse liver cells by nitrogen oxide free radicals]. AB - The ESR signal of nitrosyl complexes appears in mouse liver after hydroxylamine injection in vivo. This ESR signal testifies to the appearance of NO free radicals. The concomitant accumulation of [3H]uridine in liver RNA evidences for stimulation of transcription. This stimulation is about the same order of magnitude as the stimulation of transcription after gamma-irradiation of mice with lethal doses. It is assumed that NO and, possibly, other oxygen free radicals can disorder the gene expression machinery in the cells. PMID- 3036258 TI - [Changes in superoxide dismutase activity in the presence of electron donors and acceptors]. AB - The activity of superoxide dismutase (SOD) from bovine erythrocytes was measured by the inhibition of nitrotetrazolium blue reduction rate in superoxide anion radical generation systems--xanthine/xanthine oxidase of NADH/phenazine methasulfate. The enzyme activity increases in the presence of compounds acting as electron donors in radical-involving reactions and decreased in the presence of compounds possessing the properties of electron acceptors. Activation of SOD by electron donors and its inhibition by electron acceptors was dependent on the concentration of the above compounds. In the absence of SOD electron donors and acceptors did not change the rate of tetrazolium blue reduction by superoxide anion radicals. The role of the new type of SOD regulation for the enzyme functioning in the cell is discussed. PMID- 3036259 TI - Obsessive-compulsive disorder: psychobiological approaches to diagnosis, treatment, and pathophysiology. AB - The diagnosis, treatment, and pathophysiology of obsessive-compulsive disorder (OCD) were examined in a series of studies utilizing psychobiological approaches. Putative biological markers previously reported in depression were studied in this disorder and revealed that on some measures [Dexamethasone Suppression Test and rapid eye movement (REM) latency on sleep electroencephalogram (EEG)], OCD patients resemble those with major depressive disorder (MDD), whereas on others [REM density, platelet serotonin uptake, probably platelet 3H-imipramine binding, and 5-hydroxy-indoleacetic acid (5-HIAA) in cerebral spinal fluid (CSF)] they do not. The relationship between OCD and MDD was further explored in a double-blind, randomized crossover study designed to compare the antiobsessional effects of two tricyclic antidepressants, clomipramine (CMI) and desipramine (DMI), in a nondepressed cohort of OCD patients. CMI was found to have significant antiobsessional effects in this group, whereas in the same patients, DMI lacked therapeutic effects. These results suggest that not all antidepressants are antiobsessive and that some property of CMI, such as its potent serotonergic effects, may be of pathophysiological relevance for OCD. The role of serotonin in this disorder was then tested using the pharmacological challenge strategy. A novel serotonin postsynaptic receptor (5HT-1) agonist, m-chlorophenylpiperazine (m-CPP), was administered orally (0.5 mg/kg) under double-blind, placebo controlled conditions to OCD patients and controls. In addition, a serotonergic receptor antagonist, metergoline (4 mg), was given to a subset of OCD patients. Relative to healthy volunteers, the OCD patients became significantly more anxious, depressed, and dysphoric after m-CPP administration. Moreover, in the OCD patients, obsessive-compulsive symptoms increased markedly after m-CPP and decreased significantly following metergoline administration. These results demonstrate that agents that bind to the 5HT-1 receptor can acutely affect the symptoms of OCD patients. The striking behavioral effects of these direct postsynaptic receptor ligands and the relative specificity of clomipramine as an antiobsessional agent suggest that serotonergic neurons may play a role in the pathophysiology, as well as mediating the pharmacological reduction, of obsessional symptoms. PMID- 3036260 TI - Anisomycin inhibition of estradiol-induced and progesterone-facilitated luteinizing hormone surges in the immature rat. AB - These studies were designed to examine the effect of anisomycin, a potent and reversible inhibitor of protein synthesis with low systemic toxicity in rodents, on induction of luteinizing hormone (LH) surges by estradiol and their facilitation by progesterone. Immature female rats that received estradiol implants at 0900 h on Day 28 had LH surges approximately 32 h later (1700 h on Day 29). Insertion of progesterone capsules 24 h after estradiol led to premature (by 1400 h) and enhanced LH secretion. Protein synthesis was inhibited by 97%, 95%, 47%, and 16% in the hypothalamus-preoptic area (HPOA) and by 98%, 87%, 35%, and 0% in the pituitary at 30 min, 2 h, 4 h, and 6 h after s.c. injection of anisomycin (10 mg/kg BW), respectively. A single injection of anisomycin at 0, 3, 6, 9, 12, 24, 27, or 30 h after estradiol treatment significantly lowered serum LH levels at 32 h. The effect of injecting anisomycin at 0, 24, or 27 h was overridden by progesterone treatment at 24 h, but LH secretion was delayed serum LH levels were basal (10-30 ng/ml) at 1400 h but elevated (500-800 ng/ml) at 1700 h. Complete suppression of LH surges in estradiol-plus-progesterone-treated rats was achieved with 2 injections of anisomycin on Day 29 at 0900 h and again at 1200 h or 1400 h. Further experiments were designed to examine proteins that might be involved in anisomycin blockade of progesterone-facilitated LH surges. Intrapituitary LH concentrations at 1700 h on Day 29 were 70-80% higher (102 +/- 12.5 micrograms/pituitary) in rats that received 2 injections of anisomycin than in vehicle-treated controls (58.5 +/- 7.7 micrograms/pituitary). There were no significant effects of anisomycin on cytosol progestin receptors in the HPOA (7.1 +/- 1.5 fmol/tissue, anisomycin; 7.2 +/- 0.3, vehicle) or pituitary (8.3 +/- 1.3 fmol/tissue, anisomycin; 11.7 +/- 2.9, vehicle) at this time. The concentration of pituitary gonadotropin-releasing hormone receptors (GnRH-R), however, was significantly lower after anisomycin (265 +/- 30 vs. 365 +/- 37 fmol/mg protein) treatment. These results suggest that both estradiol-induced and progesterone facilitated LH surges involve protein synthetic steps extending over many hours. Blockade of progesterone-facilitated LH surges by anisomycin appears to be due primarily to an effect on release of LH to which lowering of GnRH-R levels may contribute. PMID- 3036261 TI - Absence of specific luteinizing hormone-releasing hormone (LHRH) receptors in the ovaries of Djungarian hamsters (Phodopus sungorus). AB - High affinity binding sites for luteinizing hormone-releasing hormone (LHRH) were characterized in Djungarian hamsters. Scatchard analysis was used to demonstrate specific LHRH-binding in hamster and, serving as controls, rat pituitaries (dissociation constant, KD = 0.6 nM, binding capacity, BM = 2.5 +/- 0.7 fmol/mg tissue; KD = 0.6 nM, BM = 6.9 +/- 1.9 fmol/mg tissue, respectively). In contrast to results obtained with rat ovaries (KD = 0.9 nM, BM = 3.0 +/- 0.9 fmol/mg tissue), no specific LHRH-binding was detected in hamster ovaries. Thus, it seems that direct gonadal action of LHRH in the Djungarian hamster is not involved in ovarian regulation. PMID- 3036262 TI - Characterization of gonadotropin-releasing hormone (GnRH) binding to pituitary receptors in goldfish (Carassius auratus). AB - Goldfish pituitary gonadotropin-releasing hormone (GnRH) receptors were characterized by using a superagonist analog of teleost GnRH (tGnRH-A; [D-Arg6, Trp7, Leu8, Pro9-NHEt]-GnRH). Equilibrium binding of 125I-tGnRH-A to a goldfish pituitary membrane preparation was achieved after a 30-min incubation at 4 degrees C; binding was significantly reduced after increasing incubation temperature to 22 degrees C. Binding of the radioligand was a function of tissue concentration, with a linear correlation over the range of 0.5-2 pituitary per tube. Incubation of the pituitary membrane preparation with increasing concentrations of 125I-tGnRH-A indicated saturable binding at radioligand concentrations of 470 pM and above. The binding of 125I-tGnRH-A was found to be reversible after addition of the cold analog, and the dissociation curve could be resolved into two linear components; slower rates of dissociation of 125I-tGnRH-A were observed after the addition of excess unlabeled tGnRH than after the addition of tGnRH-A, indicating that the analog is more effective in displacing the label than the native peptide. Addition of the cold analog displaced bound 125I-GnRH-A, and Scatchard analysis suggested the presence of at least two classes of binding sites: a high-affinity/low-capacity site and a low affinity/high-capacity site. Bound 125I-GnRH-A was displaced by tGnRH from both sites in parallel to that observed with tGnRH-A, indicating that both peptides bind to the same classes of binding sites; however, tGnRH-A had a greater affinity for the receptors than the native tGnRH. These results demonstrated the presence and provided characterization of GnRH receptors in goldfish pituitary. PMID- 3036263 TI - The effect of human chorionic gonadotropin, dibutyryl cyclic adenosine 3',5' monophosphate, prostaglandins, and 25-hydroxycholesterol on acute progesterone secretion by dissociated rabbit luteal cells in vitro: evidence for independent effect of human chorionic gonadotropins and lipoproteins. AB - Although estradiol is the established luteotropic hormone in the rabbit, the corpus luteum also contains a luteinizing hormone (LH)-activated adenylate cyclase system and cyclic adenosine 3',5'-monophosphate (cAMP)-dependent protein kinase, which suggests that LH and cAMP may play a physiological role in regulating luteal progesterone production. The present study examined whether human chorionic gonadotropin (hCG) and cAMP derivatives stimulate progesterone production by dispersed rabbit luteal cells in static and perifusion incubations. Results of this study show that progesterone production by rabbit luteal cells is significantly stimulated (p less than 0.05) by hCG concentrations at or greater than 0.1 IU/ml or by dibutyryl cAMP concentrations at or greater than 5 mM. Both agents produce maximal stimulations of approximately 4-fold. However, neither prostaglandin E2 or F2 alpha at concentrations of 0.1-3.0 micrograms/ml altered progesterone secretion. When luteal cells were incubated with maximal concentrations of hCG and lipoproteins together, the resultant progesterone secretion was additive. This suggests that the effects of hCG and lipoprotein are independent. Both responses could be blocked completely by cycloheximide (10(-4) M), and thus appear to be dependent on protein synthesis. The cholesterol derivative 25-hydroxycholesterol (20 micrograms/ml) partially overcame the steroidogenic block by cycloheximide, suggesting that transport of cholesterol, regardless of its origin, into mitochondria was an essential protein-mediated event in these cells. Inhibition of the side-chain-cleavage enzyme by aminoglutethamide blocked progesterone production by rabbit luteal cells in vitro. Although estradiol may dominate in the regulation of luteal progesterone production physiologically, this study clearly demonstrates that potential mechanisms do exist in the rabbit corpus luteum for cAMP-mediated stimulation of progesterone production in the rabbit. PMID- 3036264 TI - The adenylate kinase reaction acts as a frequency filter towards fluctuations of ATP utilization in the cell. AB - The buffering ability of the adenylate kinase reaction with respect to the phosphate potential and efficiency of oxidative phosphorylation in the presence of a fluctuating load conductance were studied by computer simulations. Fluctuations of the load conductance, i.e., of the irreversible ATP-utilizing reactions in the cell, were generated by integrating an Ornstein-Uhlenbeck diffusion process. This real or colored noise was then injected into the set of differential equations describing the rate laws for the changes of the adenine nucleotide concentrations based on a simple nonequilibrium thermodynamic model of oxidative phosphorylation. Numerical integration of this system of stochastic differential equations allowed us to investigate the influence of different parameters on the performance of this energy converter. Probability density estimates revealed that the variance of the efficiency about its optimal value was significantly reduced by the adenylate kinase reaction. It was found that the buffering ability of this enzyme is restricted to a specific frequency domain of the fluctuations of the load conductance. This frequency filtering was confirmed by substituting the random fluctuations of the load conductance by simple sinusoidal perturbations. All these studies revealed that for each domain of frequencies of the load perturbations there exists an optimal activity of the adenylate kinase which minimizes deviations from optimal efficiency of oxidative phosphorylation. PMID- 3036265 TI - Electrophoretic mobility of lambda phage HIND III and HAE III DNA fragments in agarose gels: a detailed study. PMID- 3036266 TI - Benzodiazepine receptor binding: influence of physiologic and pharmacologic factors. PMID- 3036267 TI - [Delayed development of hypertension and increased aortic relaxation in spontaneously hypertensive rats after neonatal sympathectomy]. AB - The effect of neonatal sympathectomy on vasodilator responses to acetylcholine (ACh) and cAMP has been studied in aortic rings of spontaneously hypertensive rats (SHR) and normotensive animals. The relaxation of intact SHR aorta in response to ACh and cAMP was 20-35% lower than that of normotensive rats. Sympathectomy in normotensive rats did not affect the level of blood pressure and aorta reactivity to Ach. In SHR, sympathectomy caused a decrease in blood pressure, while relaxation in response to ACh and cAMP increased, as compared to intact SHR, but remained lower than in normotensive rats. The data obtained suggest that the decrease in arterial pressure of sympathectomized SHR is a result not only of the reduction in sympathetic effects but also of the increase in smooth muscle relaxation. PMID- 3036268 TI - [The role of cAMP in generating excitatory serotonin responses of neurons in the edible snail]. AB - The neurons of the dorsal surface of snail Helix subesophageal ganglia respond similarly to the application of serotonin and the intracellular cAMP injection. These responses represent membrane depolarization. They increase in amplitude with membrane hyperpolarization and have a reverse potential between +10 and -30 mV. Presumably, these responses are associated with increased conductance for several ions. The values of the reverse potentials of serotonin and cAMP responses coincide in 7 out of 17 cells. Phosphodiesterase inhibitor theophylline caused a reversible increase in the amplitude and duration of both serotonin and cAMP responses and, used at a concentration of 1 mM, simulated them. The results obtained meet 2 out of 4 criteria demonstrating that cyclic nucleotides mediate a neurotransmitter response. It is suggested that cAMP may act as a second messenger in excitatory serotonin responses of snail Helix neurons. PMID- 3036270 TI - [Independent benzodiazepine and beta-carboline binding sites in the brain of aggressive and anxious-defensive mice]. AB - Distribution of specific 3H-flunitrazepam and 3H-beta-carboline-3-carboxylate binding sites in the brain regions of aggressive and timid-defensive mice was investigated before and after subchronic injection of diazepam (5 mg/kg). The absence of differences between the affinity and concentration of 3H-flunitrazepam binding sites in diencephalon and brain cortex in aggressive and defensive mice may be explained by general benzodiazepine receptor reaction on isolation and agonistic interaction stress. Significant predominance of 3H-beta-carboline-3 carboxylate binding sites in the brain cortex, as compared to the concentration of 3H-flunitrazepam binding sites suggests the presence of specific binding sites for beta-carbolines, which have specific distribution in the brain. PMID- 3036269 TI - [Antiabstinent action of fenibut and baclofen on a model of abstinence induced by the benzodiazepine receptor antagonist CGS 8216 in rats receiving diazepam]. AB - Benzodiazepine receptor antagonist CGS 8216 (2.5 mg/kg) induced clear-cut signs of abstinence lasting 1-1.5 hours in rats after discontinuation of diazepam treatment (10-20 mg/kg/day, 20 days). Diazepam (10-20 mg/kg) and GABAB-receptor agonists: phenibut (10-100 mg/kg) and baclofen (1.25-5.0 mg/kg) abolished the signs of abstinence. Conversely, GABAB-receptor agonist THIP enhanced the signs of abstinence. It is suggested that anti-abstinence effect of phenibut and baclofen may reflect their tranquilizing activity. PMID- 3036271 TI - [Effect of hydrocortisone on superoxide radical production by phagocytosing spleen cells]. AB - Superoxide radical production by mouse phagocytic spleen cells was shown to be essentially increased 2 hours following intraperitoneal injection of hydrocortisone acetate at a dose of 50 mg/kg body weight. The addition of hydrocortisone to the suspension of mouse spleen cells has resulted in linear dependence of hormone concentration in the incubation medium on the maximum rate of superoxide radical production. The mechanism of hydrocortisone stimulating effect on the activity of plasma membrane-located NAD(P)H-oxidase of phagocytic cells is being discussed. PMID- 3036272 TI - [Monoclonal antibodies to the angiotensin-converting enzyme from the human lung]. AB - The hybridoma producing monoclonal antibody (IgG1) to human angiotensin converting enzyme (ACE) has been prepared by fusion of murine myeloma P3O1 with spleen cells of BALB/c mice immunised with a purified human lung ACE preparation. A high specificity of monoclonal antibody (MAb) binding to immobilized ACE has been demonstrated by ELISA; that of soluble ACE--by immunoadsorption test. The latter technique permits the use of impure ACE preparations for the screening procedure. This MAb did not effect ACE activity. This antibody is believed useful not only for immunoassay and immunopurification of ACE, but also as a tool for investigation of enzyme distribution in tissue as well as for studying the structure and mechanism of ACE action. PMID- 3036274 TI - [Ultrastructural localization of thiamine pyrophosphatase activity in the epitheliocytes of duodenal biopsies in peptic ulcer]. AB - Using electron microscopic studies the location of thiamine pyrophosphatase (TPPase) activity in the epithelial cells of duodenal mucosa was investigated in 39 patients with ulcer exacerbation and during the disease remission. The biopsy material was studied before and after the administration of physiological and therapeutic doses of thiamine. TPPase activity was detected on the membranes of Golgi complex, lateral cell membrane and microvilli. The possibility of intra- and extracellular localization of TPPase activity and the activity of various enzymes depending on the mechanism of thiamine absorption across the epithelial barrier is discussed. PMID- 3036273 TI - [Characteristics of beta-adrenoreceptors of L-1210 leukemia and its sarcolysine resistant variant]. AB - Beta-adrenoceptors have been discovered on the surface of cells of leukaemia L1210 and its variant resistant to sarcolysine (D, L-melphalan). One type of functionally active receptors with dissociation constant Kd for L-[3H] dihydroalprenolol about 0.02 nM and 360 receptors per cell have been revealed in leukaemia L1210 cells. In the resistant cells two types of functionally inactive receptors with Kd1 approximately 0.02 nM (420 receptors per cell) and Kd2 approximately 2.5 nM (3000 receptors per cell) have been revealed. This property of beta-adrenoceptors may be one of the causes of tumour cell resistance to sarcolysine. PMID- 3036275 TI - [A method of detecting hybridoma-produced antibodies to cell nucleus endonucleases]. PMID- 3036276 TI - Arginyl-glycyl-aspartic acid sequences and fibrinogen binding to platelets. AB - Human fibrinogen has an Arg-Gly-Asp-Ser (RGDS) sequence at residues 572-575 of its A alpha-chain. Although RGDS-containing peptides inhibit fibrinogen binding to stimulated platelets, these peptides also inhibit platelet binding of human fibrinogen fragment X and rat fibrinogen, which lack RGDS sequences corresponding to A alpha 572-575. Thus competition between free RGD-containing peptides and internal RGDS sequence at A alpha 572-575 is not the basis for their inhibition of fibrinogen binding to platelets. Addition of a Thr to the carboxy-terminus and an Asn to the amino-terminus of the RGDS sequence, the amino acids corresponding to A alpha 576 and 571 respectively, reduced the inhibitory potency of RGDS containing peptides by fourfold to tenfold. Arg-Gly-Asp-Phe (RGDF) corresponds to A alpha 95-98, and the RGDF peptide was an effective inhibitor of fibrinogen binding, fourfold to fivefold more potent than RGDS. Thus, local primary structure may play an important role in regulating the capacity of RGD sequences in proteins to interact with specific adhesion receptors. PMID- 3036277 TI - Lactoferrin binding by leukemia cell lines. AB - Monocytes and macrophages have receptors for the iron-binding protein lactoferrin. Lactoferrin acts as a potent inhibitor of granulocyte-macrophage colony stimulating factor production when it binds to these cells. Using a rosette assay and immunofluorescence, we have shown that cultured leukemia cells, including the human erythroid leukemia cell line K562, also have lactoferrin binding sites. The number of binding sites on K562 cells was estimated using soluble 59Fe-lactoferrin. Inhibition studies demonstrate that lactoferrin binding sites are distinct and unrelated to receptors for transferrin or the Fc portion of IgG, which are present on K562 cells. However, electrostatic forces may be important for lactoferrin binding, since other polycationic proteins (eg, protamine) inhibit lactoferrin binding. Prior treatment of K562 cells with trypsin nearly abolishes lactoferrin binding. However, these cells recover their ability to bind lactoferrin when trypsin is removed. Unlike transferrin receptors, the expression of lactoferrin binding sites is not regulated by cellular iron status. Cytosine arabinoside arrests the proliferation of K562 cells and simultaneously leads to a reduction in lactoferrin surface binding, suggesting that lactoferrin binding may be dependent on cell proliferation. PMID- 3036278 TI - Clinical significance of low levels of myeloperoxidase positivity in childhood acute nonlymphoblastic leukemia. AB - The clinical significance of a low percentage of myeloperoxidase-positive blast cells in childhood acute nonlymphoblastic leukemia was determined. Of 155 consecutive cases studied by cytochemical staining methods, 14 were characterized by 4% to 15% (median 6%) myeloperoxidase-positive blasts. All 14 cases showed reactivity to Sudan black B stain, and 7 had Auer rods. The morphological subtypes of leukemia were M1 (8 cases), M2 (3), M4 (1), and M5 (2). Immunological marker studies disclosed the lymphoid-associated T11 antigen on cells from 8 of the 11 cases tested. Other lymphoid-related findings in these 8 cases included the T3 antigen and E rosette formation in 1 case each. Among cases that were prospectively studied for the expression of lymphoid-associated markers, 6 of 8 with low levels of myeloperoxidase positivity compared with only 1 of 44 with higher levels (greater than 15%) possessed such features (P less than 0.001). We conclude that low levels of myeloperoxidase reactivity distinguish cases of acute leukemia in which the blast cells coexpress lymphoid (T11 antigen) and myeloid markers. PMID- 3036279 TI - Changes in neutrophil surface protein composition accompany phagocytosis. AB - Phagocytosis is a critical host defense mechanism of neutrophils. In this study, membrane protein changes occurring during phagocytosis were studied in human neutrophils using surface radiolabeling before or after phagocytosis of various target particles. Cells were labeled at the cell surface using lactoperoxidase catalyzed iodination or neuraminidase-galactose oxidase-NaB3H4, galactose oxidase NaB3H4, or periodate-NaB3H4 techniques. Such studies are complicated by the fact that these techniques identify many surface proteins on the phagocyte, and labeling after phagocytosis occurs often results in radiolabeling proteins of the target particle, thus making changes in cell-surface proteins more difficult to detect. Immunoprecipitation with monoclonal antibody AHN-1, which reacts with a carbohydrate present on several human neutrophil surface proteins and inhibits phagocytosis, eliminated interference caused by radiolabeled proteins of the target particle and simplified analysis by restricting the study to a limited number of proteins. AHN-1 immunoprecipitated less radiolabeled protein from neutrophils labeled after phagocytosis of particles opsonized with IgG or complement than from cells labeled before phagocytosis. Isolation of phagocytic vesicles containing opsonized emulsified paraffin oil demonstrated that three proteins of mol wt 105,000, 140,000, and 170,000 recognized by AHN-1 were internalized in the phagocytic vesicle during phagocytosis. PMID- 3036280 TI - Is presynaptic modulation of norepinephrine release altered in the mesenteric vasculature of adult spontaneously hypertensive rats? AB - There is a substantial body of indirect pharmacologic evidence which has been interpreted as indicating that presynaptic receptor-mediated modulation of noradrenergic vascular neurotransmission is altered in the mesenteric vasculature of SHR. Enhanced release of either total 3H or 3H-NE after prelabeling with 3H-NE has been demonstrated to occur from the mesenteric vasculature of SHR after administration of isoproterenol or angiotensin II. However, evidence has now been obtained in the mesenteric vasculature demonstrating that there are no differences in the release of endogenous NE from adult SHR preparations versus that from adult WKY preparations. PMID- 3036281 TI - Actions of parathyroid hormone in the cardiovascular system. AB - Recent studies from several laboratories have suggested that the cardiovascular system may be considered a new target organ system for parathyroid hormone (PTH). PTH caused a sharp, dose-dependent reduction in blood pressure of several mammalian models. This hypotensive response was mediated by direct interaction of PTH with the vascular smooth muscle of arteries and resistance vascular beds. Increases in intracellular cyclic AMP concentrations were temporally and quantitatively correlated with smooth muscle relaxation by PTH. Direct (nonreflex) positive inotropic and chronotropic effects have been observed in cardiac tissue after treatment with PTH. Considering these and other observations, a physiological role for PTH as a homeostatic regulator of the cardiovascular system may be speculated. PMID- 3036282 TI - Modulation of vascular tone by atrial natriuretic factor. AB - Synthetic atrial natriuretic factor (ANF) has been found to be a potent relaxant of isolated vascular preparations. ANF is more effective in relaxing agonist induced versus depolarized contractile events and the relaxation efficacy appears inversely related to the level of contractile preload. The vasorelaxation to ANF is associated with elevations of cyclic GMP via an activation of the particulate form of guanylate cyclase. Both species and regional-vascular differences exist, in vitro as well as in vivo, in the vasorelaxation response to ANF which may reflect an altered distribution of high affinity receptors and/or different states of active vascular tone of the particular preparation. PMID- 3036283 TI - Evidence for endothelium-dependent vasodilation of resistance vessels by acetylcholine. AB - In the perfused mesenteric arterial vasculature of the rabbit, vasodilation by acetylcholine (ACh) was almost completely blocked after a 15-min perfusion of the vasculature with 0.2% collagenase, an enzyme capable of removing endothelial cells. In the perfused mesenteric arterial vasculature of the rat, vasodilation by ACh was markedly, though not completely, inhibited by hemoglobin (10 microM), an agent which can inactivate endothelium-derived relaxing factor (EDRF). These results suggest that a major component of vasodilation of mesenteric resistance vessels in rabbit and rat by ACh is mediated by EDRF. PMID- 3036284 TI - Local control of blood pressure by purines. AB - Dual control of local blood flow by purines is described: adenosine 5' triphosphate (ATP) released as a cotransmitter with noradrenaline from perivascular sympathetic nerves acts on P2X-purinoceptors on smooth muscle cells to produce vasoconstriction; ATP released from endothelial cells during hypoxia (and ADP released from aggregating platelets) acts on P2Y-purinoceptors on endothelial cells which results in production of endothelium-derived relaxing factor and subsequent vasodilatation. It is suggested that the endothelial mediated vasodilatation is a pathophysiological mechanism to protect the host tissue (e.g. brain or heart) from damage produced by hypoxia following ischaemia. The ATP released from the endothelial cells is rapidly broken down to adenosine which augments this protective mechanism by acting directly on P1-purinoceptors on vascular smooth muscle to produce a longer lasting component of vasodilatation and on perivascular sympathetic nerve terminals to inhibit release of excitatory neurotransmitters. The possibility that impairment of normal endothelial-mediated responses in atherosclerosis and hypertension can lead to local vasospasm is considered. PMID- 3036286 TI - Estrogen sulfates: biological and ultrastructural responses and metabolism in MCF 7 human breast cancer cells. AB - The biological effects and ultrastructural alterations by different estrogen-3 sulfates (E1-3-S and E2-3-S) and estradiol-17-sulfate (E2-17-S) were studied in the MCF-7 mammary cancer cell line in culture. The estrogen-3-sulfates very significantly stimulated the progesterone receptor (PR). The values (in pmoles/mg DNA +/- SE) were: control, 0.46 +/- 0.09; E1-3-S, 2.24 +/- 0.30, and E2-3-S, 2.56 +/- 0.45. The value of PR after E2-17-S incubation (0.56 +/- 0.24) was similar to the non-treated cells. The PR values obtained by the incubation of unconjugated estrone and estradiol were: 2.63 +/- 0.45 and 2.27 +/- 0.36, respectively. Analysis of the unconjugated estrogens in the medium indicated significant hydrolysis of estrogen-3-sulfates but not of E2-17-S. Using [3H]-E1-3-S, an important transformation was observed inside the cells, a great part being converted to estradiol (greater than 60% in the nuclear fraction). Electron microscopic examination indicated alterations in the secretory system after incubation with estrogen-3-sulfates similar to those obtained with unconjugated estradiol. The effect provoked by E2-17-S was significantly less than for the other sulfates. As estrogen sulfates are quantitatively the most important form of estrogens in the mammary gland, it is suggested that estrogen-3-sulfates play an important role in the biological responses to estrogens in breast cancer. PMID- 3036285 TI - Role of monocytes in the inhibitory effect of calcitriol on PHA-stimulated lymphocytes. AB - The possible role played by monocytes in the inhibitory effect of calcitriol on phytohemagglutinin (PHA)-stimulated lymphocyte proliferation was assessed by testing the effect of this sterol under different cell culture conditions. Calcitriol had a dose-dependent inhibitory effect on lymphocyte proliferation in concentrations ranging from 10(-10) up to 10(-8) M. The effect of 10(-9) M calcitriol was almost completely abolished by: a) monocyte depletion, b) inhibition of prostaglandin (PG) synthesis by indomethacin, and c) addition of exogenous interleukin-2 (IL-2). These results suggested that the inhibitory effect of calcitriol was mediated through monocytes. This possibility was substantiated by the following observations: a) the calcitriol inhibitory effect was restored when autologous adherent cells were added to monocyte-depleted PBM cells; b) the supernatant of adherent cells cultured for 24 hours in the presence of calcitriol exerted a marked inhibitory effect on lymphocyte proliferation; and c) this effect was not longer evident when adherent cells were cultured in the presence of calcitriol plus indomethacin. These data support the hypothesis that calcitriol acts, at least partially, through the monocytes, inducing an increased release of PG, with subsequent inhibition of IL-2 synthesis, then resulting in a decreased lymphocyte proliferation. PMID- 3036287 TI - [Current aspects of diseases due to papillomaviruses]. PMID- 3036289 TI - Influence of derivation on the lipophilicity and inhibitory actions of cardiac glycosides on myocardial Na+-K+-ATPase. AB - Lipophilicity and inhibitory actions on guinea-pig heart Na+-K+-ATPase of twenty six digitalis and six strophanthus glycosides comprising the aglycones, mono-, bis-, tris-sugar, alkylated (acylated) tris-sugar, acyl steroid derivatives and three cardanolides were investigated. Their octanol/water partition coefficients (P), reversed phase thin layer (r.t.l.c.) and reversed phase high performance liquid chromatography (r.h.p.l.c.) were determined and the viability of these methods as a measure of the lipophilicity of the cardiotonic steroids evaluated. The influence of lipophilicity and so also structural changes on the inhibitory effects of the cardiac glycosides on myocardial Na+-K+-ATPase was then examined. It is concluded that (a) r.t.l.c. and r.h.p.l.c. are just as effective as the conventional shake-flask method for estimation of the lipophilicity of cardiac glycosides and (b) the inhibitory potencies of cardiotonic steroids on the myocardial Na+-K+-ATPase increase with growing lipophilicity. The relationship between these two parameters is, however, governed by the influence of substitution or derivation of structural components on their inhibitory potencies on the myocardial Na+-K+-ATPase. PMID- 3036288 TI - The role of calcium in the cyclic AMP response to histamine in rabbit cerebral cortical slices. AB - The effect of calcium on the H1- and H2-receptor components of the cyclic AMP response to histamine in rabbit cerebral cortical slices has been investigated. Removal of calcium ions from the incubation medium during the preparation, preincubation and final incubation of brain slices significantly reduced the cyclic AMP responses to adenosine, histamine and the H2-selective agonist, impromidine. Removal of calcium ions from the incubation medium during only the final incubation with agonists did not influence the responses to adenosine, histamine, impromidine and the H1-selective agonist, 2-thiazolylethylamine. Final incubation of rabbit cerebral cortical slices in calcium-free buffer containing EGTA (1 mM) however, selectively reduced the cyclic AMP responses to the H1 agonists histamine and 2-thiazolylethylamine without affecting the response to impromidine or adenosine. These latter incubation conditions significantly reduced the maximal extent of the augmentation of impromidine- or adenosine stimulated cyclic AMP accumulation produced by H1-receptor stimulation, without affecting the EC50 values of the H1-agonists. Calcium-free/EGTA conditions did not, however, alter the dose-response parameters for the response to the H2 agonist, impromidine. These data provide further evidence that the two histamine receptor systems affect cyclic AMP accumulation in rabbit cerebral cortical slices by different mechanisms. PMID- 3036290 TI - Adenomatous polyposis: an association with carcinoma of the thyroid. AB - A review of the St Mark's Hospital Polyposis Registry has revealed an association between adenomatous polyposis (familial polyposis coli) and thyroid carcinoma. Even though full clinical information was unavailable on all patients in the registry, it is evident that young women (below 35 years of age) are at particular risk of developing thyroid cancer, mainly of a papillary type, their chances of being affected being approximately 160 times that of normal individuals. All patients with adenomatous polyposis should thus have regular thyroid examination. PMID- 3036291 TI - Controlled trial of gamma linolenic acid in Duke's C colorectal cancer. PMID- 3036292 TI - ABC of AIDS. AIDS and the lung. PMID- 3036293 TI - Insulinoma unmasked by the Cambridge diet. PMID- 3036294 TI - ABC of AIDS. Neurological manifestations. PMID- 3036295 TI - Endocrine response to different doses of ACTH in cows. PMID- 3036296 TI - Plasma oestrone sulphate and progesterone concentrations in cows and ewes associated with fetal death and abortion. PMID- 3036298 TI - Golgi study of the isocortex in an insectivore: the common European mole (Talpa europaea). AB - In the common European mole (Talpa europaea) the isocortex is a six-layered structure representing about 50% of the total cerebral cortex. The internal granular layer is narrow in the occipital region, however, probably reflecting a poorly developed visual system in an animal adapted to life in a subterranean environment. Golgi impregnation of projection cells and most local-circuit neurons of layers III-VI suggests a relatively well-developed isocortex in this insectivore. The presence of extraverted neurons in the so-called accentuated layer II and the amount of local-circuit neurons with very long beaded dendrites, however, probably represents primitive characteristics of the isocortex in mammals. PMID- 3036297 TI - Brain stem auditory nuclei and their connections in a carnivorous marsupial, the northern native cat (Dasyurus hallucatus). AB - The cytoarchitecture and connections of the brain stem auditory nuclei in the marsupial native cat (Dasyurus hallucatus) were studied using Nissl material in conjunction with the retrograde transport of horseradish peroxidase injected into the inferior colliculus. Some features different from those of Eutheria include the disposition of the cochlear nuclear complex medial to the restiform body, a lack of large spherical cells in the anteroventral cochlear nucleus, a small medial superior olive, and a large superior paraolivary nucleus. PMID- 3036299 TI - Cerebellar corticonuclear and corticovestibular fibers from the posterior lobe of the albino rat, with comments on zones. AB - The topographic organization of cerebellar cortical efferent fibers from the posterior lobe of the albino rat was studied by silver impregnation methods. The corticonuclear fibers from the vermis of the posterior lobe project to the caudomedial and middle parts of the medial cerebellar nucleus (MN) with medio lateral localization; fibers from the medial portion of the vermis project to the caudomedial part of the MN (MNcm) and those from the lateral portion project to the middle part of the MN (MNm). Corticovestibular fibers originate in the caudal vermis lateral to the area projecting to the MNcm and MNm, and terminate in the dorsal part of the lateral vestibular nucleus (LVN). The origins of corticonuclear fibers to the dorsolateral protuberance of the MN (MNdlp), the posterior interpositus nucleus (PIN) and the anterior interpositus nucleus (AIN) are located latero-medially in the intermediate cortex. Fibers from the area adjacent to the vermis and rostral to the prepyramidal fissure project to the MNdlp, while those from the area lateral to the origin of the corticovestibular fibers and caudal to the fissure project to the medial AIN. Together, these areas comprise a medial portion of the intermediate cortex. Corticonuclear fibers to the PIN and lateral AIN originate from the lateral portion of the intermediate cortex. The corticonuclear fibers to the dorsolateral hump and lateral nucleus originate from the medial and lateral portions of the lateral cortex, respectively. PMID- 3036300 TI - Antidromic response to medullary pyramid stimulation in rats and its relation to that in cats. AB - The response evoked in the cerebral cortex of laboratory rats after stimulation of the medullary pyramid is surface-positive. It begins 0.9-1.6 ms after the stimulus, attains peak amplitude (up to 2 mV) in 0.8-1.2 ms and lasts 2-4 ms. It occurs throughout the anterior two-thirds of the dorsal cortex and is largest lateral to bregma, with a secondary maximum in the somatosensory area II. Although it depends on antidromic conduction in pyramidal tract fibers for its production, it varies in amplitude, configuration and latency at different recording sites and at the same sites on repeated trials. It reverses polarity deep in the cortex to become a large, negative wave deep in layer V, and maintains that polarity into the white matter. Current source density analysis reveals a strong sink in layer V, with a strong source just superficial to that sink and a weaker source in layer VI. The antidromic response disappears during spreading depression, but recovers more rapidly than the primary response evoked by skin stimulation. It decreases progressively in amplitude with continuous 200 Hz iterative stimulation, and recovers slowly at the end of stimulation. The primary response evoked by contralateral forepaw and hindpaw stimulation is highly localized, being entirely within the antidromic response distribution. The antidromic response in laboratory rats consists of a small, surface-positive component analogous to the pure antidromic response of cats, and of a large, surface-positive response analogous to that found in woodchucks, rabbits, opossums and slow lorises. It is argued that this latter response results from synaptic action in pyramidal tract axon collaterals, probably onto cells in layer V, rather than being a purely antidromic event. PMID- 3036302 TI - Localization and characterization of atrial natriuretic peptide binding sites in discrete areas of rat brain and pituitary gland by quantitative autoradiography. AB - Atrial natriuretic peptide [rat (r) ANP6-33 or ANP99-126] binding sites were localized in discrete areas of rat brain and pituitary gland using quantitative autoradiographic techniques. High numbers of rANP6-33 binding sites were concentrated in the circumventricular organs (the organon vasculosum laminae terminalis, organon subfornicalis, and area postrema) and selected hypothalamic nuclei (the nucleus supraopticus, nucleus preopticus medianus and nucleus paraventricularis). High binding was also present in the choroid plexus and the bulbi olfactorii (laminae medullaris interna). A relatively low number of rANP6 33 binding sites was observed in other olfactory, limbic and brainstem areas (the nucleus tractus solitarii, nucleus motoris dorsalis vagii and nucleus hypoglossi), the eminentia mediana and the pituitary gland (anterior and posterior lobes). High-affinity rANP6-33 binding sites were demonstrated in the organon subfornicalis and the area postrema after incubation of consecutive sections from individual rat brains with 125I-rANP6-33 in concentrations from 20 to 400 pM. rANP6-33 binding sites were concentrated in areas associated with angiotensin II and/or vasopressin, suggesting an interaction among these peptides in the central nervous system. PMID- 3036301 TI - Separation distress in infant rhesus monkeys: effects of diazepam and Ro 15-1788. AB - Disruption of the primate mother-infant attachment bond is a naturally occurring stressor that results in marked behavioral, physiological, and endocrine activation. We studied the effect that altering benzodiazepine systems has on the behavioral and endocrine response of infant rhesus monkeys (1-27 weeks of age) to brief separation from their mothers. In the first experiment, the benzodiazepine agonist diazepam (0.1 and 1.0 mg/kg) significantly increased locomotion and social behavior and decreased inactivity and distress vocalizations in infant monkeys undergoing separation. In the second experiment, the benzodiazepine antagonist Ro 15-1788 (5 and 10 mg/kg) had no significant effects on the infants' separation response. In the third experiment, administration of diazepam 1.0 mg/kg was followed by administration of Ro 15-1788 10 mg/kg in infants undergoing separation. Ro 15-1788 blocked the decreases both in inactivity and in plasma ACTH and cortisol concentrations caused by diazepam, suggesting that these effects are mediated through benzodiazepine receptors. These data support the hypothesis that in primates, endogenous benzodiazepine systems modulate the behavioral and endocrine response to the naturally occurring stress of separation. PMID- 3036303 TI - Different postnatal ontogeny of two [3H]neurotensin binding sites in rat brain. AB - Two distinct neurotensin binding sites have been identified in rat brain: the NT1 acceptor site (levocabastine-sensitive) and the NT2-receptor site. In rat forebrain, NT2-receptors were present at birth, revealed a maximal level (13.8 fmol/mg tissue) on day 10 of postnatal life but a much lower plateau (3.0 fmol/mg tissue) in adult rats. NT1-acceptors were not detected before day 10 and became maximal at day 30. It is suggested that the loss of NT2-receptor sites during the postnatal development may be the expression of the regression of a transient redundancy of neuronal connections. PMID- 3036304 TI - Interrelation between MEPP amplitude and MEPP frequency in different regions along the frog neuromuscular junction. AB - Experiments were done to study the relationship between miniature endplate potential (MEPP) frequency and MEPP amplitude in different regions along the frog neuromuscular junction (NMJ). The position of the release site producing a MEPP and the amplitude of a MEPP at its release site were evaluated by using the spatial decay method after simultaneous intracellular recordings at each distal end of the NMJ. The NMJ was divided in 10 regions of equal length. It was found that MEPP amplitude and MEPP frequency are smaller in distal regions than in proximal ones. Moreover, we observed a logarithmic relationship between MEPP frequency of a given region and the mean MEPP amplitude of the same region. These results suggest that the probability to produce a MEPP and the quantity of neurotransmitter liberated are two synaptic functions controlled or affected by common mechanisms. PMID- 3036306 TI - Ontogenetic development of peripheral benzodiazepine binding sites in rat brain, heart and lung. AB - In rats, the brain exhibits negligible ontogenetic changes, while a steady marked increase in the density of peripheral benzodiazepine binding sites (PBS) has been demonstrated in the heart and lungs, reaching maximal levels at 31 days after birth. It may be that PBS play a role in the cellular proliferation of these peripheral organs. The present findings are consistent with current evidence of a relationship between these binding sites and mitochondrial function. PMID- 3036305 TI - The role of gamma-aminobutyric acid mechanisms of the zona incerta-lateral hypothalamus in the catalepsy and muscle rigidity evoked by morphine. AB - Picrotoxin or bicuculline were injected bilaterally into the zona incerta-lateral hypothalamus (ZI-LH) of the rat. Each drug (50 ng) inhibited or abolished the catalepsy induced by 20 mg/kg s.c. of morphine. Each drug also strongly inhibited the tonic electromyographic activity (EMG) induced by 10 mg/kg s.c. of morphine in the gastrocnemius soleus muscle (GS). The obtained results demonstrate participation of the ZI-LH in both catalepsy and rigidity induced by a systemic administration of morphine. PMID- 3036307 TI - Brain Na+,K+-ATPase activity possibly regulated by a specific serotonin receptor. AB - Na+,K+-ATPase activity in 6 regions of adult brain was measured after incubation with varying concentrations of serotonin. A concentration-dependent increase in enzyme activity was observed in 4 regions, with cerebral cortex and cerebellum showing the largest response. These results together with previous ones suggest that serotonin modulates brain Na+,K+-ATPase activity through a specific receptor located in target neurons or glial cells. PMID- 3036308 TI - Electrophysiological studies on the specificity of the cholecystokinin antagonist proglumide. AB - Recent evidence suggests that the glutaramic acid derivative proglumide (PROG) is a selective antagonist of cholecystokinin (CCK) in the rat CNS. The extent of this selectivity has now been characterized in more detail. Iontophoretic or intravenous (i.v.) administration of PROG was ineffective against the excitatory effect of iontophoretically applied neurotensin on midbrain dopamine (DA) cells, the excitatory effect of substance P and the inhibitory effect of Met-enkephalin on prefrontal cortical neurons, and the inhibitory effect of histamine on neurons of the sensorimotor cortex. In contrast, PROG blocked the excitatory effect of the C-terminal octapeptide of CCK in all 3 areas. Furthermore, iontophoretic PROG diminished, whereas CCK enhanced the inhibitory effect of similarly applied DA and GABA on DA cells. PROG pretreatment (1 mg/kg, i.v.) reduced the inhibitory potency and maximum effect of i.v. apomorphine (APO) on A9 DA neurons, while the inhibitory potency of APO was enhanced by i.v. CCK. Pretreatment with PROG plus CCK resulted in APO effects which were no different from those after PROG alone. Chronic treatment with PROG (1 mg/kg, p.o., 21 days) resulted in a return to normal of DA cell APO sensitivity. Combined, these findings suggest that PROG may be a relatively selective CCK antagonist, that the functional effect of dendritically released DA may be influenced by endogenously released CCK, and that tolerance may develop to the effects of chronic CCK receptor blockade. PMID- 3036309 TI - Active sodium-potassium transports in skeletal muscles of deoxycorticosterone hypertensive rats. AB - CNS-induced suppressions of active Na+, K+ transport was investigated in both 'tonic' muscles, soleus (SOL), and 'twitch' muscle, extensor digitorum longus (EDL) of deoxycorticosterone acetate (DOCA) hypertensive rats. There was a marked K+ loss and Na+ accumulation in the skeletal and smooth muscles of DOCA hypertensive rats. The cellular K+ loss was in the order of SOL greater than EDL greater than diaphragm greater than intestine greater than aorta. However, liver, kidney and CNS organs such as cerebrum, cerebellum and medulla oblongata were spared from this K+ fall. Sciatic nerve sectioning or cervical transection activated the active Na+, K+ transport in SOL during DOCA hypertension but inhibited further the pump activity in EDL. The application of tetrodotoxin on the sciatic nerve also activated the Na+, K+ transport in SOL but inhibited the transport in EDL. The facilitatory effect of denervation on the pump activity in SOL was abolished by pretreatment with ouabain. Injection of curare had no effect on Na+ and K+ contents in both SOL and EDL. These results indicate that the CNS is involved differently on the neural regulations of the active Na+, K+ transport systems in SOL and EDL of DOCA hypertensive rats. PMID- 3036310 TI - Functional properties of the cerebellorubral synapses in the cat. AB - Facilitation of cerebellorubral transmission was illustrated in nembutalized cats on paired stimulation of the cerebellar nucleus interpositus by an example of monosynaptic EPSPs of the rubrospinal neurons. It was found that the facilitation stated is not determined by the change in the presynaptic volley of impulses. There is reason to believe that the facilitation is determined by specific features of functioning of the cerebellar synapses on the red nucleus neurons. PMID- 3036311 TI - In botulinum type A-poisoned frog motor endings ouabain induces phasic transmitter release through Na+-Ca2+ exchange. AB - Ouabain (100 microM) applied for 60 min to botulinum A (BoTx) poisoned motor junctions increases, in a time-dependent manner, the mean number of acetylcholine quanta released by nerve stimulation and enhances the delayed transmitter release. The drug does not affect spontaneous quantal release. The observed effects on evoked transmitter release cannot be explained by changes in the configuration of presynaptic currents recorded from motor terminals. They suggest that in BoTx-poisoned motor endings the level of intraterminal Ca2+, lower than that required for the activation of quantal transmitter release, can be effectively increased through the reversed operation of an Na+-Ca2+ exchange system that normally uses the Na+ gradient to extrude Ca2+. PMID- 3036312 TI - Comparison of binding sites for the calcitonin gene-related peptides I and II in man. AB - Specific and saturable binding sites of the synthetic human calcitonin gene related peptide-I (CGRP-I) and -II (CGRP-II) have been identified in membrane homogenates of the human central nervous system, heart and spleen. Half maximal inhibition of the binding of 125I-CGRP-II was achieved with 0.25 nM and 1.7 nM of non-radioactive CGRP-I or -II, and with 260 and 850 nM salmon calcitonin in the spinal cord and cerebellum, respectively, but 1 microM human calcitonin was not recognized. Autoradiographs of diencephalic sections revealed different distribution of 125I-CGRP-I and -II binding sites. PMID- 3036313 TI - Further evidence for endogenous hypothalamic serotonergic neurons involved in the cimetidine-induced prolactin release in the rat. AB - The aim of the present work was to further explore the possible relationship between the prolactin-releasing effect of cimetidine and hypothalamic serotonergic neurons controlling pituitary hormone secretion. In a first approach, the prolactin-releasing effect of the drug was determined in adult male rats with total deafferentation of the hypothalamus. Cimetidine injection (60 mg/kg) produced a significant rise in prolactin, but not in luteinizing hormone (LH), both in deafferented rat and in sham-operated controls; by 15 min there was a 5-6 fold increase in prolactin titers. Methysergide, a serotonin receptor blocker, used in a dose (2.5 mg/kg), route (i.p.) and time (50 min earlier) which did not modify the hormone basal level in rats with total deafferentation of the hypothalamus, was able to prevent completely the prolactin release evoked by cimetidine. The same preventive effect on prolactin release was observed with the serotonin receptor blocker ketanserin (5 mg/kg, i.p., 30 min earlier). It is concluded that the prolactin-releasing effect of cimetidine is located at a hypothalamic level related to serotonergic neurons. PMID- 3036314 TI - Drinking-induced alterations in reward pathways: an in vivo autoradiographic analysis. AB - An in vivo autoradiographic technique permitted the visualization of discrete neuroanatomical changes in opiate receptor binding as a result of 23-h water deprivation and drinking. Two groups of rats (n = 5) were placed on a 23-h water deprivation schedule for 10 days. On the last day, one group was given access to water for 15 min. These groups, plus a matched ad libitum water control group (n = 5), received an injection of 0.002 mg/kg [3H]diprenorphine ([3H]Dpr) through chronically implanted jugular catheters followed by preparation for opiate receptor autoradiography. Relative cerebral blood flow was estimated non quantitatively by the injection of 75 microCi/kg iodo-[14C]antipyrene into 3 additional groups identically treated. Results indicated that water-deprivation stress increased [3H]Dpr binding in the claustrum, lateral hypothalamus, amygdala and ventral tegmental area while decreasing binding in the medial frontal cortex, lateral septum, dorsolateral thalamus and central gray. All effects of water deprivation were reversed in animals receiving water. Observations of changes in relative blood flow were shown to have no correlation with changes in opiate receptor binding. It appears that water deprivation stress causes a reduction in opioid release in areas along the mesotelencephalic dopamine pathway which may contribute to a drive state. Water intake may then reduce or otherwise alter the drive state through the release of opioids along these pathways, contributing to the perception of reward. PMID- 3036315 TI - Intensive anti-oxidant pretreatment retards motor nerve degeneration. AB - Intensive pretreatment of cats with a combination of the antioxidants D-alpha tocopherol (200 IU) and selenium (50 micrograms) once daily for 5 days (p.o.) was found to significantly preserve the functional capacity of degenerating soleus motor nerve terminals (measured at 48 h after axon section at the hip) in the in vivo soleus nerve-muscle preparation. The preservation of function was apparent in terms of: a greater soleus contractile response to nerve stimulation at low frequencies, a more rapid recovery from D-tubocurarine-induced neuromuscular block, and a better maintenance of tetanic contractile tension during high frequency nerve stimulation. The ability of antioxidants to retard the anterograde axonal degeneration (i.e. 'Wallerian') process suggests that lipid peroxidation may be a fundamental mechanism of neuronal degeneration. PMID- 3036316 TI - The effect of the imidazodiazepine Ro 15-4513 on the anticonvulsant effects of diazepam, sodium pentobarbital and ethanol. AB - The ability of the imidazodiazepine Ro 15-4513 to antagonize the anticonvulsant effects of diazepam, sodium pentobarbital and ethanol was investigated. Ro 15 4513 alone significantly lowered seizure threshold to bicuculline and this effect subtracted from the anticonvulsant effects of sodium pentobarbital and ethanol. In contrast, Ro 15-4513 completely reversed the anticonvulsant effects of diazepam, consistent with suggestions that it is a competitive ligand for benzodiazepine receptors. PMID- 3036317 TI - Opioid receptor subtypes in the rat spinal cord: electrophysiological studies with mu- and delta-opioid receptor agonists in the control of nociception. AB - We have compared the ability of selective mu- and delta-opiate agonists to modulate nociceptive transmission at the level of the rat dorsal horn using electrophysiological approaches. Single-unit extracellular recordings were made from neurones in the lumbar dorsal horn of the intact rat under halothane anaesthesia. Neurones could be activated by both A- and C-fibre electrical stimulation (and by natural innocuous and noxious stimuli). Agonists were applied directly onto the cord in a volume of 50 microliters. The intrathecal administration of 3 agonists, Tyr-D-Ala-Gly-MePhe-Gly-ol (DAGO) (mu-selective) (2 X 10(-3)-10 nmol) Tyr-D-Thr-Gly-Phe-Leu-Thr (DTLET) (mu/delta) (7 X 10(-4)-70 nmol), and cyclic Tyr-D-Pen-Gly-Phe-D-Pen (DPDPE) (delta) (2 X 10(-2)-100 nmol) produced dose-dependent inhibitions of C-fibre-evoked neuronal activity whilst A fibre activity was relatively unchanged. DAGO produced near-maximal inhibitions which could be completely reversed by naloxone (1.5 nmol) whilst DPDPE causes less marked inhibitions which could only be partially reversed by naloxone (1.5 13.5 nmol). DTLET produced effects intermediate to those of DAGO and DPDPE. The results suggest that both mu- and delta-opioid receptors can modulate the transmission of nociceptive information in the rat spinal cord. PMID- 3036318 TI - Characteristics of CA1 activation through the hippocampal trisynaptic pathway in the unanaesthetized rat. AB - The hippocampal CA1 field is activated by the entorhinal cortex mainly through the hippocampal excitatory trisynaptic circuit. Field responses of the CA1 region were evoked by ipsilateral CA3 or perforant path volley (mono- or trisynaptic activation, respectively) in paralyzed, locally anaesthetized rats and studied as a function of the stimulus patterns presented. The relationship of these responses with the concomitant EEG was also explored. Results showed that mono- and especially trisynaptically evoked responses were progressively enhanced by increasing the stimulus frequency from 0.1 to 1.0 Hz. At specific intensities the trisynaptically evoked population spike (PS) was present only with a rather fixed frequency of stimulation (approximately 0.5 Hz). PS was produced in 100% of the responses using 0.7 Hz, indicating the existence of a threshold-like level for this stimulus parameter. The frequency of presented paired pulses differentially affected pair-pulse facilitation of mono- and trisynaptically evoked excitatory postsynaptic potentials (EPSP): higher frequency decreased the former and increased the latter. All evoked responses studied (i.e. EPSP and PS) showed steep increments and decrements in amplitude, clearly developing several clusters. Moreover, the amplitude distribution of trisynaptic PS often varied spontaneously from maximal to negligible values, showing an all-or-none distribution. Clustering was interpreted as evidence of the existence in the hippocampus of functional neuronal aggregates. All-or-none distribution of trisynaptic PS was found to be associated with the EEG pattern, PS amplitude being maximal during irregular EEG activity and minimal during theta rhythm. Present results suggest that (1) the entorhinal cortex may exert modulatory actions on CA1 by a mechanism widely based on the frequency of the input; (2) information transfer from the entorhinal cortex to other brain areas throughout the hippocampus is biased by hippocampal EEG; and (3) electronic coupling may be functionally predominant in the hippocampus. PMID- 3036319 TI - Cerebellar Na+,K+-ATPase activity is increased during early postnatal development of the estrogenized female rat. AB - The activities of Na+,K+-ATPase and Mg2+-ATPase were measured in the crude P2 synaptosomal fraction of the cerebellum through age 35 days in female rats injected s.c. with 500 micrograms estradiol benzoate 24 h after birth. Estrogenization did not affect Mg2+-ATPase. However, the activity of Na+,K+ ATPase was significantly increased above control values between ages 5 and 20 days. These data demonstrate an age-dependent estrogen-induced effect on cerebellar Na+,K+-ATPase during early postnatal development. PMID- 3036320 TI - [Effect of ACTH injection and graft of bursa of Fabricius on the adrenocortical activity of bursectomized chicks at 80 hours of incubation]. AB - Early embryonic bursectomy (BFX) disturbed the adrenocortical functioning. The stress-unresponsive period that occurred in controls, and lasted for 2-3 weeks after hatching, no longer appeared in BFX chicks. In contrast, the magnitude of the stress-induced hypercorticosteronemia was much lower in BFX than in sham operated 5 week-old chicken. It was assumed that such adrenocortical dysfunction was due to bursal deprivation, since grafting bursal buds onto the chorio allantoic membrane of BFX embryos restored all the parameters under study, i.e., the post-hatching stress unresponsive period and the high magnitude of stress induced responses in adults. Factor(s) involved in such interregulation are not known but do not seem to affect directly adrenocortical cells because intramuscular injection of a moderate dose of ACTH resulted in the same hypercorticosteronemia whether 3 day-old and 5 week-old chicks had been bursectomized or sham-operated. PMID- 3036321 TI - [Endonuclease III of Escherichia coli and the repair of oxidized thymines and AP sites]. AB - Escherichia coli endonuclease III is not an endonuclease. It breaks the C3'-O-P bond 3' to an AP site in DNA by catalysing a beta-elimination and not a hydrolysis. Therefore, it is a phosphoric monoester-lyase. PMID- 3036322 TI - [A minority of thymic lymphomas induced in the rat by a murine radioleukosis virus presents insertion in the vicinity of c-myc and a majority, a new nonviral polyadenylated RNA]. AB - Thymic lymphomas were induced in rats either with the cell culture-propagated radiation leukemia virus complex, RadLV/VL3, or with a molecularly cloned isolate, RadLV/VL3 (T + L +). Four of thirty lymphomas, that were examined for rearrangements of the c-myc domain, displayed alterations in the vicinity of the c-myc gene, compatible with the idea of proviral integration. One of the tumours was investigated further, and was shown to contain a full length-proviral insert upstream of c-myc. Eight of nine lymphomas, that were investigated with respect to RNA expression, contained a novel polyadenylated RNA which could be detected with a molecular probe derived from the U5 portion of the retroviral long terminal repeat, but not with probes derived from the U3 portion or from the whole retroviral genome. These findings suggest that a RadLV/VL3 (T + L +) provirus can induce or activate RNA synthesis from c-myc by an enhancer mechanism, and from another cellular gene by a promotion mechanism. PMID- 3036323 TI - [Relative biological effectiveness (RBE) of neutrons produced by 50 MeV deuterons and by 34, 45, 65 and 75 MeV protons]. AB - RBE of p(34) + Be, p(45) + Be, p(65), + Be, p(75) + Be and d(50) + Be neutron beams produced at the cyclotron "Cyclone" of Louvain-la-Neuve were measured. The biological criterion was the regeneration of the crypts of the intestinal mucosa (50 regenerated crypts per circumference) after abdominal irradiation in mice. Taking the p(65) + Be neutrons as reference, RBE values were found equal to 1.12, 1.07, 1.00 (Ref.), 0.96 and 1.02 respectively. These results are consistent with those published for cell lethality in vitro. However, the RBE variation is smaller than this previously obtained in the laboratory for growth inhibition in Vicia faba. PMID- 3036324 TI - Pathophysiological aspects of myocardial hypertrophy. AB - Heart hypertrophy in response to increased workload is a complex process in which this organ adapts to the environment by increasing the muscle mass in terms of additional contractile units and formation of different types of contractile proteins (myosin isozymes). In addition, augmentation of membrane function with respect to calcium transport activities of sarcolemma and sarcoplasmic reticulum occurs at early stages of cardiac hypertrophy associated with hyperfunction of the myocardium. However, if cardiac hypertrophy is left unattended beyond a certain period, physiological hypertrophy is converted to pathological hypertrophy whereby the cardiac muscle is unable to generate an adequate amount of contractile activity. It appears that the sympathetic nervous system is activated for producing beneficial effects at early stages but an elevated level of sympathetic tone for a prolonged period could result in dysfunction of the cardiac muscle. The transition of physiological hypertrophy to pathological hypertrophy seems to be due to the occurrence of intracellular calcium overload in the myocardial cell as a consequence of defects in the membrane calcium transport systems. It is suggested that careful attention should be paid not only to removal of the stimulus responsible for cardiac hypertrophy but also to lowering sympathetic tone. Efforts should also be made to prevent the occurrence of intracellular calcium overload due to membrane defects. PMID- 3036325 TI - Some aspects of the regulation of pyruvate kinase levels in Neurospora crassa. AB - Pyruvate kinase levels were monitored in Neurospora crassa mycelium (grown on different carbon sources for varying time intervals) by immunoprecipitation using polyclonal antibodies raised against a purified enzyme preparation. Pyruvate kinase specific mRNA was demonstrated by hybridization of Northern and dot blots of total RNA with a N. crassa pyruvate kinase gene fragment. Two pyruvate kinase specific mRNA species were detected in mycelia of all ages examined. An age dependent and carbon source dependent variation in the pyruvate kinase protein and mRNA levels was encountered: both registered an increase for up to about 20 h and a subsequent decline; growth on acetate and sucrose resulted in significantly higher yields of both, relative to that on medium containing ethanol and alanine. Stress caused by heat shock depressed the pyruvate kinase mRNA levels. PMID- 3036327 TI - Family history of colorectal cancer as a marker of potential malignancy within a screening program. AB - Epidemiologic studies have shown that asymptomatic adult relatives of colorectal cancer patients are at increased risk for developing this tumor. A prospective, published pilot study confirmed this added risk and demonstrated the importance of the family history of cancer as a marker of potential malignancy. The study group was enlarged to include 471 asymptomatic adult, first degree relatives of patients having large bowel neoplasia (cancer or adenomatous polyps) but without polyposis syndromes. These first degree relatives were screened by fecal occult blood examinations and flexible sigmoidoscopy, followed by colonoscopy when indicated. Adenomatous polyps or cancer were found in 8.1% of the study group as compared with 3.7% in a comparison group of screens, not having the same family history of neoplasia and undergoing similar screening tests. Of the study group the age-adjusted rate for colorectal adenomas or cancer increased threefold (P less than 0.001) for subjects older than 40 years and an even higher fivefold relative risk was found for large bowel cancer only (P = 0.01). This was true even if there was only one relative with colorectal neoplasia (P less than 0.01) but was even more pronounced among those having more than one affected relative. The results confirm the usefulness of the family history, of even one member with large bowel neoplasia, in isolating a group at high risk for these lesions. This group would most likely benefit from regular cancer and adenomatous polyp screening particularly when older than 40 years. PMID- 3036329 TI - Evaluation of gamma-enolase as a tumor marker for lung cancer. AB - The alpha-enolase and gamma-enolase in tumor tissues and sera of patients with lung cancer were determined with an enzyme immunoassay system. Tissue gamma enolase in small cell carcinoma of the lung (SCCL, n = 11), large cell carcinoma (n = 11), and non-SCCL (except for large cell carcinoma) (n = 34) were enhanced approximately 35-fold, ninefold, and fourfold, respectively; tissue gamma/alpha + gamma value of SCCL was significantly higher than that of normal lung tissue (P less than 0.01). Serum gamma-enolase level was elevated (greater than 6.0 ng/ml) in 14/18, 3/10, and 15/60 patients with SCCL, large cell carcinoma, and non-SCCL (except for large cell carcinoma), respectively, and serum gamma/alpha + gamma value of SCCL was significantly higher than that of healthy subjects (P less than 0.01). Immunohistochemically, the gamma-enolase was positive in 29/31 of the lung cancers. Serum gamma-enolase value is a useful tumor marker for staging and monitoring treatment of patients with lung cancer, and serum gamma/alpha + gamma value may be useful for differential diagnosis of SCCL from non-SCCL or in differentiating lung cancers possessing neuroendocrine features from other lung cancers. PMID- 3036326 TI - Evidence for a glucose effect on N-acetylglucosamine catabolism in Candida albicans. AB - Two strains of Candida albicans were examined for a glucose effect on the catabolism of N-acetylglucosamine. It was shown that the induction of N acetylglucosamine uptake capacity was almost completely blocked by glucose at 0.5% (w/v), whereas that of N-acetylglucosamine kinase was partially repressed. PMID- 3036328 TI - Expression of ABH and Lewis blood group antigens in combined hepatocellular cholangiocarcinoma. Possible evidence for the hepatocellular origin of combined hepatocellular-cholangiocarcinoma. AB - Expression of ABH, Lewis, and sialyl Lea antigens was studied in five combined hepatocellular-cholangiocarcinomas. Formalin-fixed liver tissues were immunostained for those antigens using well-characterized monoclonal antibodies and an avidin-biotin-peroxidase complex (ABC) method. Results were compared with those obtained in normal liver tissues and cholangiocarcinomas, and also with the previous observations of the authors on hepatocellular carcinomas. Although not detected in normal parenchymal liver cells, A, H, Lewis, and sialyl Lea antigens were found in combined hepatocellular-cholangiocarcinoma cells. Incompatible A antigen also was detected in one blood type O patient. Distribution and intensity of the antigens were similar to those in hepatocellular carcinomas and different from those in cholangiocarcinomas. No preferential accumulation of blood-group antigens could be found in the area of cholangiocarcinoma-like differentiation of the combined hepatocellular-cholangiocarcinoma. The observations suggested that Regional morphological differentiation of the hepatocellular-cholangiocarcinoma might not be always associated with the change in the expression of the blood group antigens. Moreover, the expression was essentially the same between the hepatocellular-cholangiocarcinoma and the typical hepatocellular carcinoma. The hepatocellular-cholangiocarcinoma, therefore could be a variant of the hepatocellular carcinoma. PMID- 3036331 TI - Biological markers in urologic cancer. AB - Tumor markers (TMs) play an important part in the management of urologic cancer. Alpha-fetoprotein, human chorionic gonadotropin, and occasionally lactic dehydrogenase serological determinations have become indispensable in the management of nonseminomatous germ cell testicular tumor patients, particularly after initial therapy, whereas human chorionic gonadotropin and probably placental alkaline phosphatase are important in seminoma. Prostatic acid phosphatase has long been important for the monitoring of patients with carcinoma of the prostate. The availability of the immunologic assays instead of the enzymatic assays has improved sensitivity somewhat but clinical interpretation has also become more complicated. Prostatic specific antigen is already an important tissue marker for carcinoma of the prostate and promises to be an important serological one, possibly surpassing prostatic acid phosphatase in importance. Analysis of DNA by automated flow cytometry is becoming important in the early detection and follow-up of bladder cancer patients. Studies concerning the tissue analysis of blood group antigens in bladder cancer continue to demonstrate that this approach can provide unique clinical information and interesting biological insights, but its role in routine clinical management remains to be determined. Currently, TMs have little clinical significance in renal cell carcinoma, but the availability of monoclonal antibodies to renal cell carcinoma preferential antigens may change this deficiency soon. In fact, in the near future, monoclonal antibodies will probably reveal many new substances for many urological cancers which can be used for markers serologically, histochemically, and, with their corresponding antibody, for radioimmune imaging and possibly immunotherapy. Now, as then, familiarity with the nuances of the marker and good clinical judgement will be essential. PMID- 3036330 TI - Hepatitis B related childhood hepatocellular carcinoma. Childhood hepatic malignancies. AB - The case of hepatocellular carcinoma (HCC) with foci of hepatoblastoma in a 7 year-old boy, the son of a hepatitis B surface antigen (HBsAg) carrier mother, is described. Twelve other malignant liver tumors in children were tumors in children were also reviewed for HBsAg and hepatitis B core antigen (HBcAg). Both were negative in all (nine) hepatoblastomas. One of three HCC demonstrated positivity for HBsAg. These cases illustrate the importance of hepatitis B virus infection in early childhood and stress the need for careful screening in pregnant women, irrespective of ethnic backgrounds. PMID- 3036332 TI - Selecting initial therapy for pediatric genitourinary cancers. AB - A few decades ago, there were few choices in the initial management of children with genitourinary tumors. Radical surgical removal was the only line of attack that promised any chance of survival. Improvement in the results of multimodal therapy in the last 15 years have radically altered the outlook for these children, hence the choice of therapy. As with other childhood cancers, the choice of therapy is based on risk-benefit evaluations of the roles of surgery, irradiation, and chemotherapy, since all three modalities have their associated morbidities. Current emphases are on preservation of function without compromising cure. The large cooperative clinical trials have emphasized this aspect of pediatric oncology. They have demonstrated, for example, that radiation therapy can be omitted from primary management of early stage Wilms' tumor patients who are given adequate adjuvant chemotherapy as can both radiation therapy and ablative surgery in certain cases of early stage rhabdomyosarcoma. Routine retroperitoneal node dissections have been shown to be of dubious diagnostic or therapeutic value in boys with testicular cancers. The need for bilateral oophorectomy in girls with dysgerminoma can similarly be questioned. Choices of initial therapy, therefore, are not static. They are becoming wider with each advance in multimodal therapy. Clinicians must keep abreast of the results of clinical trials so they can offer their patients the combination of treatments that will preserve function, and produce the smallest number of late complications without jeopardizing survival chances. PMID- 3036333 TI - Selecting initial therapy. Seminoma and nonseminoma. AB - About 80% of seminoma presents as low-stage disease. If clinical studies are negative, the usual initial therapy is "prophylactic" radiotherapy to the retroperitoneal zone. If clinical Stage II disease is evident, radiotherapy versus primary chemotherapy is being studied. Primary chemotherapy is treatment of choice for clinical Stage III disease. Postchemotherapy radiographic lesions are safe to follow without surgical extirpation as they are usually necrotic. Surveillance for clinical Stage I disease is another option. For nonseminoma, clinical Stage I disease has been managed with staging RPLND. But 70% of such cases will have negative nodes. Hence, primary surveillance studies are under way, with chemotherapy reserved for those who relapse clinically (estimated 95% survival). Sadly, surveillance has not been effective when applied on an ad hoc basis at the community level. Problems are compliance, delayed detection of relapse, nonreporting of failures. Clinical Stage II disease is managed with RPLND. Adjuvant, limited postoperative chemotherapy is an option versus no postoperative chemotherapy followed by full chemotherapy for those who relapse as Stage III disease later. Another option under study for Stage II disease is primary chemotherapy with RPLND surgery reserved for those who achieve only a partial remission. PMID- 3036334 TI - Sexuality and fertility in urologic cancer patients. AB - With the advent of effective treatment for urologic cancer, the preservation of sexual function and fertility has become an important goal. Some cancer treatments damage the physiological systems involved in reproduction. All have a psychological impact on sexuality. For men with prostate cancer, current issues in sexual rehabilitation include the debate on nerve-sparing radical prostatectomy, the role of vascular damage in causing erectile dysfunction after radiotherapy, and the need for a better understanding of hormonal effects on central and peripheral mechanisms of sexual function. In the treatment of men and women with bladder cancer, the sexual function morbidity of radical cystectomy is described in data from prospective interview studies. Sexual desire and orgasm remain normal after surgery despite disruption of the genital vasocongestion accompanying sexual arousal. Long-term follow-up studies of testicular cancer patients suggest that some increase in sexual dysfunction does occur. Infertility remains a concern for a subgroup of younger, childless men. Attempts to modify or eliminate retroperitoneal lymphadenectomy are discussed, as is recovery of spermatogenesis after chemotherapy and radiotherapy. Sexual function in patients with penile, urethral, or renal cell carcinoma is briefly reviewed. PMID- 3036335 TI - The treatment of advanced or recurrent malignant genitourinary tumors in children. AB - Wilms' tumor, embryonal rhabdomyosarcoma, and yolk sac tumor are the most frequently diagnosed malignant genitourinary tumors in children. The randomized therapeutic trials developed by the National Wilms' Tumor Study (NWTS) committee have produced significant improvements in the survival of Wilms' tumor patients. Those with Stage IV, favorable histology Wilms' tumor now have a 2-year survival rate of 87% to 93%. Patients with recurrent Wilms' tumor are being prospectively treated with new drugs and drug combinations with the goal of identifying active agents for inclusion in future therapeutic trials. Girls with localized embryonal rhabdomyosarcoma of the vagina have a 100% survival rate after treatment on the Intergroup Rhabdomyosarcoma Study (IRS) (IRS-1 or IRS-2). Primary treatment of patients with tumors of the prostate, bladder neck, or trigone with chemotherapy has demonstrated the need to establish local tumor control using surgery or radiation soon after diagnosis. The IRS-III currently is evaluating new drug combinations in this group of patients with locally advanced tumors. Patients with advanced yolk sac tumor have been treated using platinum-containing combination chemotherapy regimens with very encouraging results. Future research will evaluate the timing of second-look surgical procedures and the efficacy of etoposide-containing combination chemotherapy regimens. PMID- 3036336 TI - Autoimmune basis for visual paraneoplastic syndrome in patients with small cell lung carcinoma. Retinal immune deposits and ablation of retinal ganglion cells. AB - Recently, patients with visual paraneoplastic syndrome (VPS) were described, a binocular loss of vision found in patients with small cell carcinoma of the lung (SCCL). The patients have serum antibodies against a small number of discrete antigens which are shared by the retina and small cell carcinoma cells, and which are associated with cells and processes of the ganglion cell layer of the retina. Pathologic findings are presented with regard to the presence of immunoglobulins in, and the nature of the lesions in, the central nervous system of a VPS patient. The patient's blood-brain barrier was shown to be compromised, as demonstrated by the finding of high immunoglobulin levels in the cerebrospinal fluid and immune deposits in the retina. It is further shown that within the central nervous system only the retina and optic nerve show any tissue damage with the specific loss of retinal ganglion cells and their processes. The findings support the hypothesis of an autoimmune cause for this remote effect of cancer. PMID- 3036337 TI - Isoenzyme pattern of enolase in the diagnosis of neuroendocrine bronchopulmonary tumors. AB - Electrophoretic separation of enolase isoenzymes and the measurement of enolase activity were performed in 25 lung tumor extracts. In 13 neuroendocrine (NE) tumors (nine small cell lung carcinoma [SCLC], three atypical NE tumors, and one carcinoid tumor), the NE differentiation was assessed by ultrastructural determination of neurosecretory granule (NSG) density. Twelve non-NE lung tumors also were studied (three adenocarcinomas, four epidermoid, two composite, two large cell undifferentiated carcinomas, and one lymphoma). Four normal lung tissues and 1 human brain were used as controls. The gamma gamma isoenzyme was present at a high level (mean +/- SE, 12 +/- 3%) in all NE carcinomas and consistently absent in all non-NE tumors as well as in normal lung. The alpha gamma isoenzyme was found in significantly higher proportion in NE carcinomas (mean +/- SE, 29 +/- 2%) than in non-NE tumors (mean +/- SE, 8 +/- 1%) (P less than 0.0001), despite an equally high level of total enolase activity in both groups of tumor. The separation of alpha gamma and gamma gamma isoenzymes of enolase allows for the accurate diagnosis of NE tumors and NE components of atypical NE carcinomas, and the gamma gamma isoenzyme, in contrast to gamma chain detection by immunoassay, can be considered to be a specific marker in itself of NE differentiation in lung neoplasms. PMID- 3036338 TI - Leukocytosis and large cell lung cancer. A frequent association. AB - In a retrospective study of 105 patients with non-small cell lung cancer during a 5-year period, 43 had leukocytosis. In 19 of the 43 patients, no clear cut etiology for the leukocytosis was apparent and it was attributed to the tumor itself. In these 19 patients, absolute neutrophilia was detected in 13, eosinophilia was present in three, and eleven exhibited concomitant thrombocytosis. Tumor-associated leukocytosis occurred predominantly, and eosinophilia exclusively, in patients with large cell pulmonary neoplasms. These results suggest an unusual myeloproliferative stimulus in this type of cancer. It may result from tumor cell production of hemopoietic growth factors such as granulocyte-macrophage colony-stimulating activity; however, additional studies are needed to elucidate the underlying mechanism(s), and to determine whether this is a peculiar characteristic of the cells that comprise large cell undifferentiated carcinoma of the lung. PMID- 3036339 TI - Immunoenzymatic staining methods for simultaneous demonstration of chromosomes and cell surface markers. AB - We have developed immunocytochemical staining methods for the simultaneous phenotypic and karyotypic characterization of individual cells. Following a mild hypotonic pretreatment, isolated cells are cytocentrifuged on poly-L-lysine coated slides, fixed in formol buffered acetone, and subsequently labeled with monoclonal antibodies utilizing indirect immunoenzymatic staining procedures with horseradish peroxidase (HRP) or alkaline phosphatase monoclonal anti-alkaline phosphatase (APAAP) as second antibodies. Preparations are refixed consecutively in methanol and 45% acetic acid and counterstained with either "Stains-all" (HRP labeled preparations) or Giemsa (APAAP labeled preparations). C-banding or weak G banding, which allows the identification of individual chromosomes, can be induced in labeled as well as unlabeled mitotic cells by Ba(OH)2 and/or 2 X SSC treatment after refixation, respectively. Our method has been successfully tested with a variety of monoclonal antibodies against lymphoid, myeloid, erythropoietic, and thrombopoietic cell surface antigens. It is fast, allows the adjustment of the intensity of cell surface staining, and results in permanent preparations suitable for light microscopic analysis. PMID- 3036341 TI - Common cytogenetic findings in primary breast cancer. AB - Five cases of breast cancer were cytogenetically studied by G-banding, using direct tumor preparations. Chromosomes involved in aberrations, according to frequency, were #1, #11, #3, #6, #5, and #17. In all five cases there were abnormalities of chromosomes #1 and #11. In each case chromosome #1 was involved in at least two different ways. In four cases abnormalities of chromosomes #11 exhibited nonrandom involvement of band q22-23. These findings confirm the role of chromosomes #1 and #11 in breast cancer and show that band 11q22-23, which has been reported to be an inheritable fragile site and is a specific breakpoint in acute leukemia, also may be specific in a group of breast cancer. Thus, correlation of an inheritable fragile site and a malignant disease with familial incidence seems possible. PMID- 3036340 TI - Cytogenetic and molecular genetic studies of a patient with atypical lymphoid hyperplasia. AB - We have karyotyped cells from a lymph node of a patient with atypical lymphoid hyperplasia. Among other clonal chromosomal abnormalities, a t(2;19) translocation was observed with breakpoints at 2p11.2 and 19q13. The genes for transforming growth factor alpha and beta have been mapped to 2p11-p13 and 19q13, respectively, but Southern blot analysis did not reveal any alteration in the structure of these genes. Similarly, the kappa immunoglobulin gene, which maps to 2p11-p12 was not rearranged. In addition, Southern blot analysis using immunoglobulin and T-cell receptor genes as probes, did not demonstrate any clonality of either B or T cells. We propose that this patient represents an early, polyclonal stage of atypical hyperplasia. The chromosome changes observed may have been one of the etiologic factors causing this disorder. PMID- 3036342 TI - Extracolonic manifestations of familial polyposis coli. AB - The original concept of familial polyposis coli (FPC) that it is only a genetically determined premalignant disease of the colon changed in the 1950s, with the description of Gardner's syndrome. The extracolonic manifestations of osteoma and epidermoid cyst since have been shown to be only a small part of the spectrum of both benign and malignant extracolonic manifestations of the disease. The modern concept of this condition is that FPC is a genetically determined generalized growth disorder that gives rise to tumors in various parts of the body. PMID- 3036343 TI - Influence of dietary fats on cell populations of line 168 mouse mammary tumors: a morphometric and ultrastructural study. AB - The effect of dietary fat concentration and saturation on cell composition and structure of line 168 mouse mammary tumors in vivo was studied using morphometry and electron microscopy. Both the concentration and saturation of fat fed to mice had a significant influence on the volume ratio of mast cells infiltrating line 168 tumors. Tumors of mice fed diets containing a high concentration (20%) of either safflower oil (SO) or palm oil (PO) had 2-3 times the volume ratio of mast cells than mice fed diets containing a low concentration (5%) of either fat. There were no significant differences among diets with respect to other inflammatory cell populations. Mice fed either one of the high fat diets had tumor cells with inclusions that ultrastructurally, appeared to consist of lipid. Dietary fat, however, had no observable affect on cell junctions or other morphological characteristics. Greater infiltration of mast cells in tumors of mice fed high fat diets and the eventual formation of new blood capillaries may explain the decreased latency of tumor onset and enhanced growth of tumors in mice fed diets with high concentrations of fat. PMID- 3036344 TI - Enhancement of N-methyl-N'-nitro-N-nitrosoguanidine-induced DNA amplification in a Simian virus 40-transformed Chinese hamster cell line by 3-aminobenzamide. AB - A Simian virus 40-transformed Chinese hamster cell line (CO 60) amplifies integrated viral DNA sequences as a response to treatment with a variety of carcinogens. To study a possible involvement of poly(ADP-ribose) synthesis, DNA amplification was induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), an alkylating carcinogen that strongly stimulates poly(ADP-ribose) synthesis. In the presence of 3-aminobenzamide (3AB) (2 mM), a competitive inhibitor of poly(ADP ribose) polymerase, MNNG-induced amplification was increased two to six times the level induced by MNNG alone. Concomitantly, 3AB reduced cellular poly(ADP-ribose) levels and increased MNNG-induced cytotoxicity, as expected. The effect of 3AB on MNNG-induced amplification depended both on the concentration of 3AB and the duration of its presence after MNNG treatment. By contrast, 3-aminobenzoic acid, a noninhibitory structural analogue of 3AB, had no influence on amplification induced by MNNG. These data strongly suggest an involvement of poly(ADP-ribose) in the process of DNA amplification, as it is shown that inhibition of carcinogen stimulated poly(ADP-ribose) synthesis by 3AB is correlated with an enhancement of inducible DNA amplification in this cell line. PMID- 3036345 TI - Effect of phorbol ester on growth of tumors induced by Rous sarcoma virus and on pp60src kinase activity in these tumors. AB - The inoculation of Rous sarcoma virus (RSV) into chickens results in the induction of tumors which usually grow progressively for several weeks and then regress. We have found that the direct injection of phorbol myristate acetate into growing RSV-induced sarcomas resulted in accelerated tumor regression in each of 11 cases studied. We further found that treatment of cultured Rous sarcoma cells with phorbol myristate acetate for 72 h resulted in a 50% diminution in numbers of viable cells, in comparison with controls, and a 60% reduction in DNA synthesis. We also investigated alterations in the pp60src kinase activity of cultured RSV-induced tumor cells after treatment with phorbol myristate acetate and found that pp60src kinase activity was reduced to 65-95% of control levels. This inhibition of pp60src kinase was both time- and concentration-dependent. Western blot analysis, using a tumor-bearing rabbit serum, indicated that the pp60src protein was reduced by 35-65% in treated cells, while Pr76, precursor of the structural RSV-proteins, was diminished by 10-35%. In contrast, nonviral proteins such as actin were not significantly affected. PMID- 3036346 TI - Metabolism of arachidonic acid in human lung cancer cell lines. AB - The metabolism of arachidonic acid (AA) was studied in two pulmonary bronchioloalveolar-carcinoma cell lines (NCI-H322 and NCI-H358) and two small cell lung carcinoma cell lines (NCI-H69 and NCI-H128). Exogenous AA was metabolized only in the NCI-H322 and NCI-H358 cells. There was no detectable metabolism of AA in NCI-H69 or NCI-H128 cells, either in the presence or the absence of the calcium ionophore A23187. The major metabolite of AA isolated from both NCI-H322 and NCI-H358 cells was prostaglandin E2 (PGE2). Prostaglandin endoperoxide synthase activities, expressed as immunoreactive PGE2 (pmol/min/mg protein), were 10.3 +/- 0.28 (SD) and 4.8 +/- 0.48 in NCI-H358 and NCI-H322 cells, respectively. The rate of production of PGE2 by both NCI-H358 and NCI-H322 cells was linear up to 10 min. Production of PGE2 in both cell lines was dependent upon substrate concentration and was maximal above 17 microM AA. Moreover, PGE2 did not undergo further metabolism by either the NCI-H358 or the NCI-H322 cells. Aspirin (0.1 mM), a cyclooxygenase inhibitor, decreased PGE2 production by 77 and 60% in NCI-H358 and NCI-H322 cells, respectively. In the presence of exogenous AA the calcium ionophore, A23187 (20 microM), stimulated PGE2 production in NCI-H322 cells by almost 2-fold, although it did not affect PGE2 production in the NCI-H358 cells. In contrast, A23187 stimulated the endogenous production of PGE2 in both NCI-H322 and NCI-H358 cells by 4- and 9 fold respectively. In addition, both the NCI-H358 and NCI-H322 cell lines were susceptible to the cytotoxic effects of the anticancer agent mitoxantrone in both a time and concentration dependent manner. In contrast, the two cell lines lacking detectable prostaglandin synthesis activity, NCI-H69 and NCI-H128 were unaffected by treatment with mitoxantrone. These results illustrate that there are major differences in the abilities of human lung cancer cell lines to biosynthesize and release PGE2. It is conceivable that such differences might have exploitable diagnostic and/or therapeutic implications. PMID- 3036347 TI - Absence of protein kinase C in nuclei of EL4 mouse thymoma cells. AB - Since evidence indicates that phorbol ester-induced production of interleukin 2 requires transcription, we investigated the possibility that the phorbol ester receptor acts directly in the nuclei of EL4 thymoma cells. Using a procedure that minimized plasma membrane contamination (as measured by 5'-nucleotidase activity) and maintained the integrity of the double nuclear membrane, we were unable to detect specific binding of [3H]phorbol 12,13-dibutyrate in nuclei of unstimulated cells. Treatment of cells with phorbol 12,13-dibutyrate (100 nM, 37 degrees C) for up to 6 h did not cause appearance of phorbol ester binding capacity in nuclei (4 +/- 8% of homogenate value; 5'-nucleotidase activity = 10 +/- 3%) despite translocation of 40% of the cytosolic binding capacity to the plasma membrane fraction. The failure to detect nuclear binding capacity in treated cells was not due to occupation of nuclear sites with unlabeled ligand; effective exchange binding was demonstrated by recovery of total homogenate binding capacity in treated cells of 82 +/- 13% of that in untreated cells. Treatment of isolated nuclei with DNase to liberate DNA binding proteins also failed to reveal any nuclear phorbol ester binding capacity. Assay of nuclei for protein kinase C enzymatic activity gave similar negative results. These data argue strongly against a direct action of the intact phorbol ester receptor (or the phorbol ester binding fragment) in the transcriptional activation of interleukin 2 in EL4 cells but cannot rule out the possibility of a role for the catalytic fragment. PMID- 3036348 TI - Neoadjuvant chemotherapy in the combined modality approach of locally advanced nonmetastatic breast cancer. AB - We have treated 76 patients with locally advanced breast cancer, 31 with stage IIIA, 41 with stage IIIB, and 4 with stage IV disease, with primary induction chemotherapy including an attempted hormonal synchronization in 70 patients. All were treated to maximum objective clinical response before proceeding to any local therapy. Patients achieving a complete response with a negative repeat biopsy generally received radiation therapy while patients with residual disease, partial response (PR) or no change (NC) status received debulking surgery prior to radiation therapy. Regardless of response to induction chemotherapy, patients received at least 6 additional months of chemotherapy following local therapy. Initial doses of combination chemotherapy were escalated to targeted myelosuppression. The objective response rate to induction chemotherapy was 93% with 49% complete response (CR), 44% PR, and 7% NC. The median numbers of cycles of chemotherapy to achieve a CR, PR, or NC were 5, 3, and 5, respectively. Three patients who currently have PRs are still on chemotherapy with continued tumor regression. Of 37 patients achieving a CR to chemotherapy, 35 were assessed by biopsies to determine pathological evidence of response. Twenty-three of the 37 patients (62%) were proven to be complete responders with negative biopsies. Twenty-four patients have relapsed, 6 with stage IIIA, 16 with stage IIIB, and 2 with stage IV. Five patients have had locoregional relapses alone, 4 locoregional and distant, and 15 distant alone. Median time to progression is 35.9 months for stage IIIA and 34.2 months for stage IIIB. Median survival is 35.3 months for stage IIIB and is indeterminate for stage IIIA. This aggressive primary chemotherapy regimen with hormonal synchronization followed by local therapy appears to provide excellent local control and encouraging early results on systemic disease control. PMID- 3036349 TI - Vinca alkaloids in the treatment of non-small cell lung cancer. PMID- 3036350 TI - Dynamic changes of cerebrospinal fluid shunt flow in patient's daily life. AB - The shunt flow rate will be greatly influenced by the changing posture of the patient. A newly designed method of assessing shunt flow rate by isotope clearance is described and the results of phantom experiments and clinical data are presented. This method makes it possible to assess shunt flow rates in a variety of postures, such as recumbent, or head raised or as posture changes from recumbent to sitting and eventually to upright. As patients changed from the recumbent to the sitting position, shunt flow rates ceased in some cases. In cases with low flow rates in the recumbent position, shunt flow rate increased with any elevation of the upper half of the body. In many cases, flow rates increased as the patient's position changed from recumbent to sitting and then to the upright position. The results suggest that shunt flow rates vary substantially as postures alter in a patient's daily life. PMID- 3036352 TI - In vitro evaluations of pyrophosphate/copolymer/NaF as an anticalculus agent. PMID- 3036353 TI - Effects of pyrophosphate/copolymer/NaF on dental calculus and caries formation in vivo. PMID- 3036351 TI - [Effect of acetate and bicarbonate on changes in left ventricular function caused by hemodialysis. M-mode echocardiography and systolic interval study]. PMID- 3036354 TI - Pyrophosphate adsorption onto hydroxyapatite and its inhibition of crystal growth. PMID- 3036356 TI - The anticalculus effect of dentifrices containing pyrophosphate salts and sodium fluoride. PMID- 3036357 TI - Comparative clinical study of two anticalculus dentifrices. PMID- 3036355 TI - Changes in the mineral content of artificial carious lesions brushed in vivo with anticalculus dentifrices. PMID- 3036358 TI - The anticalculus efficacy of two commercially available anticalculus dentifrices. PMID- 3036359 TI - Role of receptor-binding antibodies and anti-idiotypic antihormone antibodies in immune regulation. PMID- 3036360 TI - Idiotype network and its relevance to autoimmune diseases. Functional considerations. PMID- 3036362 TI - Calmodulin-drug interaction. A fluorescence and flow dialysis study. AB - Various Ca2+-antagonists and related compounds were probed for possible anti calmodulin properties. Some of them efficiently inhibit calmodulin dependent activity (the plasma membrane Ca2+-ATPase and the cyclic nucleotide phosphodiesterase). The I50-values for the most potent inhibitors varied between 15 and 30 uM. Using fluorescence spectroscopy and flow dialysis methods the stoichiometry of the binding of some of the drugs to calmodulin has been investigated. The number of Ca2+-dependent high affinity binding sites has been studied on trypsin fragments of calmodulin. Compound 12-114 was bound with high affinity in a Ca2+-dependent way to both halves of calmodulin, compound 200-737 recognized one high affinity binding site only in the C-terminal half of the molecule, whereas compound 36-079 demanded the intact protein to be able to interact with high affinity in a Ca2+-dependent manner. PMID- 3036361 TI - Compartmentalization of cyclic AMP elevation in neurons of Aplysia californica. AB - We have measured by radioimmunoassay the amount of total, free, and bound forms of cyclic AMP (cAMP) within the abdominal ganglion and in five identified cell bodies of neurons from Aplysia californica. In the abdominal ganglion the unbound (free) cAMP levels comprised approximately 25-30% of the total cAMP content under the unstimulated condition, i.e., bathed in high-magnesium saline. Under pharmacological conditions that blocked endogenous phosphodiesterase and activated adenylate cyclase, ganglionic free cAMP levels were elevated more than fourfold, while bound cAMP levels more than doubled. Freeze-substitution techniques were employed to facilitate isolation of individual cell bodies either before or after pharmacological manipulation of cAMP levels. The basal, free cAMP content of cells R2, LP1, R15, L11, and L2-L6 was in the range of 10-40 pmol/mg of cell protein, which accounted for approximately one-half of the total cAMP content per cell body. Determinations of individual cell volumes indicated that the basal, free cAMP concentrations ranged from 1 to 6 microM. Under the same pharmacological conditions that elevated ganglionic cAMP in levels, no changes were measured in either the free or the bound forms of cAMP in isolated cell bodies. Our results indicate that the cAMP elevation was compartmentalized within the neuropilar region of the ganglion, most likely within the processes of the nerve cells. Previous results demonstrated that cAMP injections into the same Aplysia neurons studied here induced a cAMP-activated sodium current, INa (cAMP). In this report we discuss the possibility that pharmacological elevation of cAMP within neuronal processes may reach concentrations similar to those produced by cAMP injections into somata. PMID- 3036363 TI - Calcium binding by parathyroid cell plasma membranes. AB - Parathyroid hormone (PTH) secretion from parathyroid glands is controlled mainly by extracellular calcium both in vivo and in vitro. In this study, the Ca2+ binding properties of bovine parathyroid cells have been investigated on a highly purified plasma membrane preparation with flow dialysis techniques. Scatchard plot analysis of the data shows the existence of at least two different binding sites: the high affinity have an apparent Kd1 of 6.6 X 10(-5) M and a capacity (n1) of 20.1 nmol/mg protein. The low affinity sites have an apparent Kd2 of 2.6 X 10(-4) M and a capacity (n2) of 37.7 nmol/mg protein. Furthermore, at higher total Ca2+ concentrations, additional bindings with an apparent Kd3 in the millimolar range and a capacity (n3) of 118 nmol/mg protein are detectable. Neither the apparent Kd's nor n's of these calcium binding sites was affected by isotonic substitution of the medium with NaCl or KCl, while the number of binding sites increased in Choline Cl. On the other hand, the presence of 5-10 mM LiCl in isotonic Na-K medium caused a marked decrease in calcium binding from these sites. LiCl is the only monovalent cation that both in vitro and in vivo is able to enhance PTH release regardless of high or low extracellular Ca2+ concentrations. Therefore the observed displacement of calcium from calcium binding sites seems to be rather specific and could correlate with the observed enhanced PTH secretion in parathyroid cells. It is proposed that the first event in the regulation of PTH secretion by extracellular Ca2+ is the binding of Ca2+ to the low affinity, high capacity sites present on the surface of parathyroid cells and that calcium occupancy of those sites is the necessary event to initiate intracellular signals leading to inhibition of PTH secretion. PMID- 3036364 TI - Analysis of DNA surrounding the breakpoints of chromosomal translocations involving the beta T cell receptor gene in human lymphoblastic neoplasms. AB - DNA containing breakpoints of two different t(7;9) chromosomal translocations was cloned from the T lymphoblastic tumor cell lines SUP-T1 and SUP-T3. Sequence analysis of DNA from the t(7;9)(q34;q34.3) translocation of SUP-T1 revealed that chromosome 9 DNA had recombined with DNA 5' to rearranged D-J regions in the beta T cell receptor gene of chromosome 7. Restriction analysis and hybridization studies using DNA fragments cloned from the t(7;9)(q34;32) translocation of SUP T3 confirmed that beta T cell receptor DNA is also joined to the DNA of chromosome 9 in these cells. Using hybridization probes for the two breakpoints, several other cases of T lymphoblastic tumors were shown to possess DNA rearrangements near the 9q34.3 and 9q32 sites. Hybridization with the 9q34.3 probe detected multiple transcripts in SUP-T1 RNA and small amounts of larger transcripts in T cells lacking the t(7;9)(q34;q34.3) translocation. This work directly demonstrates that the beta T cell receptor locus may frequently be involved in chromosomal translocations within T lymphoblastic neoplasms. PMID- 3036365 TI - Bovine papilloma virus plasmids replicate randomly in mouse fibroblasts throughout S phase of the cell cycle. AB - Bovine papilloma virus (BPV) replicates as a multicopy nuclear plasmid in mouse fibroblasts. Using fluorescence activated cell sorting and mitotic selection procedures, we show that the replication of BPV occurs throughout S phase of the cell cycle and that replication is confined to S phase. After one round of chromosomal DNA replication, almost one quarter of BPV plasmids have replicated more than once, while a similar number of plasmids have not replicated at all. While multiple forms of BPV exist in the cell, all forms show the same pattern of replication. These results are consistent with a model in which BPV plasmids are chosen at random for replication throughout, and only during, S phase and support the view that the completion of S phase is a specifically activated event in the cell cycle rather than simply the end of one round of chromosomal DNA replication. PMID- 3036366 TI - A transcriptional repressor encoded by BPV-1 shares a common carboxy-terminal domain with the E2 transactivator. AB - A negative-acting transcriptional regulatory factor encoded by bovine papillomavirus type 1 (BPV-1) was identified. This factor inhibits BPV-1-mediated transformation of mouse C127 cells; inhibition is BPV-1-specific and occurs only when the BPV-1 transforming genes are regulated by authentic transcriptional control elements. Plasmids expressing the inhibition function also repress E2 transactivation of the BPV-1 E2-dependent enhancer, and this repression is mediated by the same cis-acting element required for E2 transactivation. Inhibition of transformation may result from down-regulation of E2-dependent viral gene expression. Analysis of cDNA expressing the inhibition/repression activities mapped the function to the 3' domain of the E2 open reading frame. The E2 open reading frame thus encodes both positive and negative transcriptional regulatory factors, and these factors share a carboxy-terminal domain. PMID- 3036367 TI - The mechanism of chromosome 14 inversion in a human T cell lymphoma. AB - The chromosome 14 inversion produces cytogenetic breakpoints at either end of the long arm of this chromosome. Previous studies have shown that a hybrid gene (designated IgT) consisting of an immunoglobulin VH gene segment and T cell receptor J alpha C alpha segments encompasses the telomeric breakpoint in SUP-T1, a cell line derived from a human T cell lymphoma. Here, we report that the centromeric breakpoint in SUP-T1 constitutes the reciprocal of a VH-J alpha join but involves gene segments different from those at the telomeric breakpoint. Therefore, chromosome inversion and IgT formation were mediated by two sequential VH-J alpha joining events. Moreover, sequences adjacent to the centromeric breakpoint detect a T-cell-specific RNA, encoded within the immunoglobulin VH locus, whose transcriptional activity may have facilitated the illegitimate VH-J alpha rearrangements. PMID- 3036368 TI - Vaccinia virus produces late mRNAs by discontinuous synthesis. AB - We describe the unusual structure of a vaccinia virus late mRNA. In these molecules, the protein-coding sequences of a major late structural polypeptide are preceded by long leader RNAs, which in some cases are thousands of nucleotides long. These sequences map to different regions of the viral genome and in one instance are separated from the late gene by more than 100 kb of DNA. Moreover, the leader sequences map either upstream or downstream of the late gene, are transcribed from either DNA strand, and are fused to the late gene coding sequence via a poly(A) stretch. This demonstrates that vaccinia virus produces late mRNAs by tagging the protein-coding sequences onto the 3' end of other RNAs. PMID- 3036369 TI - Epstein-Barr virus gp350/220 binding to the B lymphocyte C3d receptor mediates adsorption, capping, and endocytosis. AB - The type 2 complement receptor, CR2, a B lymphocyte surface glycoprotein, is known to be a component of the EBV receptor. We now demonstrate that the major EBV outer membrane glycoprotein, gp350/220, is a highly specific ligand for CR2. EBV or beads coated with purified recombinant gp350/220 adsorb to normal B lymphocytes, cap with CR2, become endocytosed into vesicles, and are released into the cytoplasm. This is the first demonstration of herpesvirus glycoprotein cell glycoprotein receptor interaction in viral adsorption and penetration. The capping of CR2 in response to virus, gp350/220-coated beads, or anti-CR2 monoclonal antibodies is associated with cocapping of surface immunoglobulin. Interaction between CR2 and surface immunoglobulin may be important in modulating the B cell activation that normally follows EBV infection or exposure to antigen. PMID- 3036370 TI - Structure of the l(2)gl gene of Drosophila and delimitation of its tumor suppressor domain. AB - We have previously cloned lethal(2)giant larvae, a tumor-suppressor gene of Drosophila that normally controls cell proliferation and/or differentiation in the optic centers of the brain and the imaginal discs. Here we describe the structure of the l(2)gl genes as determined by sequencing genomic and cDNA clones. The structure of the cDNAs indicates the use of alternative splicing, either in the 5' untranslated exons or in the 3' coding exons. Thus the gene encodes two putative proteins of 1161 and 708 amino acids, p127 and p78, respectively, differing at their C termini. A 3'-truncated l(2)gl transposon that leaves the coding sequence of p78 intact but deletes 141 residues of p127 was capable of suppressing tumor formation in l(2)gl-deficient animals. These results suggest that the putative p78 protein is effective in controlling cell proliferation and/or differentiation. PMID- 3036371 TI - Splicing of SV40 early pre-mRNA to large T and small t mRNAs utilizes different patterns of lariat branch sites. AB - To explore the mechanism and control of alternative splicing, we have characterized the products formed by splicing of SV40 early pre-mRNA in vitro and in vivo. Large T and small t mRNAs are derived from this precursor by joining alternative 5' splice sites to a single shared 3' splice site. In contrast to pre mRNAs studied previously, we have shown that splicing to large T RNA involves the utilization of multiple lariat branch sites, while small t splicing uses a single branch site. Interestingly, the predominant branch sites utilized in splicing of large T RNA in vitro were found to differ in nuclear extracts from HeLa and human 293 cells, correlated with previously observed differences in the ratio of large T to small t mRNAs produced in the two cell types. To test the significance of this correlation, we examined the products formed by splicing of an SV40 early precursor microinjected into X. laevis oocytes. Strikingly, both the pattern of branch sites used in large T splicing and the ratio of large T to small t mRNAs produced were found to be identical to those observed in 293 cells and extracts. PMID- 3036372 TI - ATP activates dnaA protein in initiating replication of plasmids bearing the origin of the E. coli chromosome. AB - ATP is bound to dnaA protein with high affinity (KD = 0.03 microM) and hydrolyzed slowly to ADP in the presence of DNA. ADP is also bound tightly to dnaA protein and exchanges with ATP very slowly. The ATP form is active in replication; the ADP form is not. A unique conformation of oriC, formed in an early initiation stage, depends on dnaA protein being in the ATP form. The subsequent entry of dnaB protein to form a prepriming complex also requires ATP binding and is blocked by bound ADP. Inasmuch as hydrolysis of ATP is far slower than these initiation reactions and since the poorly hydrolyzable analogue ATP gamma S can replace ATP, the ATP function appears to be allosteric. The extraordinary affinity of ATP for dnaA protein, its slow hydrolysis to ADP, the profound inhibition of dnaA functions by ADP, and the very slow exchange of ADP all point to a possible regulatory role for these nucleotides in the cell cycle. PMID- 3036373 TI - Three different genes in S. cerevisiae encode the catalytic subunits of the cAMP dependent protein kinase. AB - We have isolated three genes (TPK1, TPK2, and TPK3) from the yeast S. cerevisiae that encode the catalytic subunits of the cAMP-dependent protein kinase. Gene disruption experiments demonstrated that no two of the three genes are essential by themselves but at least one TPK gene is required for a cell to grow normally. Comparison of the predicted amino acid sequences of the TPK genes indicates conserved and variable domains. The carboxy-terminal 320 amino acid residues have more than 75% homology to each other and more than 50% homology to the bovine catalytic subunit. The amino-terminal regions show no homology to each other and are heterogeneous in length. The TPK1 gene carried on a multicopy plasmid can suppress both a temperature-sensitive ras2 gene and adenylate cyclase gene. PMID- 3036375 TI - Cyclic 3',5'-adenosine monophosphate modulates vascular endothelial cell migration in vitro. AB - Using a modified Boyden chamber assay, we have examined the effect of cyclic nucleotides on bovine aortic endothelial cell migration in vitro. Dibutyrl cyclic 3',5'-adenosine monophosphate (5 mM) inhibited endothelial cell random migration by 67% and inhibited fibronectin-induced chemotaxis by 75%. Agents which significantly stimulated adenylate cyclase activity in endothelial cell membranes were also effective inhibitors of endothelial cell migration. Timolol blocked both the isoproterenol-induced stimulation of adenylate cyclase and the ability of isoproterenol to inhibit endothelial cell migration. Caffeine and isoproterenol together had a greater inhibitory effect on endothelial cell motility than either alone. These data suggest that cAMP may modulate vascular endothelial cell migration in an inhibitory fashion. PMID- 3036376 TI - A comparative study on proteins released from metaphase chromosomes after digestion with restriction endonucleases and deoxyribonuclease I. AB - Extensive digestion of Chinese hamster metaphase chromosomes with Alu I, Hae III and Hinf I released up to 40 distinct chromosomal proteins. Some of the proteins released by Hae III or Hinf I were enriched in the protein moiety liberated by Alu I but several proteins released by Hae III were not released by Alu I digestion. The amount of chromosomal protein released by deoxyribonuclease I (DNase I) was comparable to that liberated by the three restriction enzymes so far tested, while only four abundant protein species were detectable in the protein moiety released by DNase I. Two of them with molecular weights of 58,000 and 50,000 were also released by the three restriction enzymes and are similar in size to those found previously in the core-like structure of histone-depleted chromosomes. PMID- 3036377 TI - The establishment of IL-2 producing cells by genetic engineering. AB - Expression plasmids containing human interleukin-2(IL-2) cDNA under the control of viral promoters (SV40 early region, MuLV LTR, HTLV-I LTR, and ASV (Y73) LTR) were introduced into TK- mouse L cells and human FL cells to establish IL-2 producing cells. The highest levels of IL-2 producing clones were obtained in TK+ mouse L cells transformed with a recombinant plasmid having MuLV LTR as a promoter, whereas transformed cells of human FL cells (G418r) were revealed to produce IL-2 at the highest level when the cells were transfected with a plasmid containing HTLV LTR as a promoter. These results suggest that these promoter/enhancer regions possess different cell specificities in gene expression. To obtain higher levels of IL-2 production using gene amplification, the hybrid plasmids containing the hamster DHFR and human IL-2 genes were constructed and transfected into DHFR- CHO cells. DHFR+ colonies produced IL-2 at about the same level as that produced by TK+ L cells transformed with the recombinants containing MuLV LTR. Selection of methotrexate-resistant cells resulted in a 5- to 30-fold increase of IL-2 production. These cells produced IL 2 stably for at least 3 months, even in the absence of methotrexate. PMID- 3036378 TI - [Occurrence of poliovirus strains with increased neurovirulence in the waste water from the city of Prague 1983-1984]. PMID- 3036374 TI - Epstein-Barr virus-transformed B cells process and present Mycobacterium tuberculosis particulate antigens to T-cell clones. AB - We have analyzed the presentation of mycobacterial antigens by Epstein-Barr virus transformed human B (EBV-B) cells to mycobacteria-specific T-cell clones and lines, and to purified resting T cells. EBV-B cells were able to process and present not only soluble forms of antigen, such as PPD and the expressate preparation of M. tuberculosis strain H37Rv, but also particulate forms of antigen, such as whole mycobacterial H37Rv or M. bovis organisms. Electron microscopy studies demonstrated the capacity of EBV-B cells to phagocytose mycobacterial cells in 18 hr and pulsing experiments confirmed that an 18-hr of incubation is required for an efficient processing and presentation of mycobacterial determinants to T cells. The processing of whole-H37Rv particulate antigen by EBV-B cells was inhibited by the lysosomotrophic compound chloroquine and by high doses of irradiation. Finally, the analysis of the presentation of soluble and particulate mycobacterial antigens by PPD-positive and PPD-negative EBV-B cell clones has shown a preferential presentation of both forms of antigen by PPD-positive EBV-B clones. PMID- 3036380 TI - Wilms' tumor: prognostic factors and survival. AB - A statistical analysis of 38 cases of Wilms' tumor treated and followed by the Pediatric Oncology Group of Cerrahpasa Medical Faculty, from 1977 to 1984, is presented. The two-year survival rate was 67% and the five-year survival rate was found to be 63%. There was a strong correlation between the stage and age of the patients and prognosis. The median age was 16 months in stages I and II of the disease and the two-year survival rate was 87%. The median age was found to be 5 years in advanced stages and the survival rate at 2 years was 43%. No correlation was found between survival and initial symptoms of the tumor or its localization. The effects of histological grade on prognosis of disease were also investigated. PMID- 3036379 TI - 4'-Epidoxorubicin plus cisplatin as first-line therapy in the treatment of small cell bronchogenic carcinoma. AB - Twenty-seven patients with small cell bronchogenic carcinoma were treated with a combination of 4'-Epidoxorubicin 60 mg/m2 and cisplatin 50 mg/m2 i.v. every 3-4 weeks. Three patients (11%) had a complete remission (CR), and 12 (44%) had a partial remission (PR) with a 55% overall remission rate. The median duration of response was 36 weeks (range 11-256+). No severe bone marrow depression was noted. The other side effects were of mild grade. Because of the "minimal aggressiveness" of this combination for the patients, the results obtained in this preliminary phase can probably be improved by the integration of other drugs in the scheme. PMID- 3036381 TI - Microbiological properties of sulbactam combined with ampicillin. AB - Several in vitro parameters of sulbactam combined with ampicillin (Sbt/Amp) were studied in order to evaluate and compare its microbiological properties with those of beta-lactamase stable beta-lactams (ceftriaxone, cefamandole, cefoxitin). The intrinsic activity of Sbt/Amp was satisfactory and comparable to that of other beta-lactams and no significant difference was observed against beta-lactamase producing or non-producing bacteria. Besides, it was determined that sulbactam, when combined with ampicillin at different ratios (1:2, 5:1), with and without preincubation (60 min) reduces the hydrolysis of ampicillin determined by ten different standard beta-lactamases. The hydrolysis rates of ampicillin become as low as those of cefoxitin, cefamandole and ceftriaxone, which are beta-lactamase stable cephalosporins. In fact, all the antibiotics under examination were slightly hydrolyzed by several beta-lactamases, but such slight hydrolysis did not affect the antibacterial activity against beta lactamase producing bacteria. About 50% of non-beta-lactamase producing bacteria (18 strains of different bacterial species) produced an inducible beta-lactamase after ten daily subcultures with sub-inhibitory concentrations of each antibiotic, but minimum inhibitory concentrations (MICs) of Sbt/Amp never increased after stimulation in contrast with other drugs whose MICs were much higher in consequence of this procedure. Finally, no spontaneous resistant mutant to Sbt/Amp was detected, but several mutants appeared in response to the other drugs. Sulbactam makes ampicillin as effective as beta-lactamase stable cephalosporins and its use does not determine an increase of resistance or selection of resistant mutants. PMID- 3036382 TI - Assessment of DNA binding of platinum-radiosensitizer complexes by inhibition of restriction enzymes. AB - A simple and rapid method has been used to compare the binding of platinum complexes to DNA, in a relatively qualitative manner. A compound bound at or near the restriction site inhibits enzymatic cleavage of DNA; inhibition of BamHI and EcoRI activity by complexes was assessed in this study using linearized pSV2-gpt plasmid. Our particular interest was in DNA binding by complexes of platinum (Pt) with known organic radiosensitizers (RS), to determine whether the Pt was able to target the RS to the DNA. Although the Pt-RS complexes investigated themselves have moderate radiosensitizing ability (like the inorganic complexes, cis- or trans-diamminedichloroplatinum(II), c- or t-DDP) none of the Pt-RS inhibit to the same extent as c- or t-DDP. However, there appears to be some correlation between enhanced radiosensitization by Pt-RS over Pt(RS)2, with the degree of Pt binding (as assessed by our assay). Our results using isolated DNA suggest that not all complexes bind well (e.g. Pt with two RS ligands), but that in certain cases (e.g. Pt with only one RS), it is possible to target the drug to the DNA. An ammine or amine ligand may be required in order to target a radiosensitizer to DNA using platinum. PMID- 3036383 TI - Photoaffinity labeling of a delta-receptor component of NG 108-15 cells with a new enkephalin analog. PMID- 3036384 TI - Inhibitors of adenosine 3',5'-cyclic monophosphate phosphodiesterase in Daphne genkwa Sieb. et Zucc. PMID- 3036385 TI - [Immunocolloidal gold labelling electron microscopy and its application in studying the morphology of viruses]. PMID- 3036387 TI - Method for the determination of 4-demethoxydaunorubicin, its quinone and hydroquinone metabolites in human plasma and urine by high-performance liquid chromatography. AB - 4-Demethoxydaunorubicin (4-DMDNR) is a new orally active analogue of daunorubicin (DNR). We have developed a high-performance liquid chromatography (HPLC) method capable of separating and identifying 4-DMDNR, five possible fluorescent quinone metabolites and three possible non-fluorescent hydroquinone metabolites. Methods are described for high-yield synthesis of reference metabolites. The limit of detection of the fluorescence assay was less than 1 ng/ml after extraction of 1 ml plasma or urine with chloroform/propan-2-ol (2:1), with coefficients of variation in k' (HPLC column capacity factors) of less than 3% throughout the day. Efficiency of the extraction method described exceeded 80% in control experiments. Blood and urine samples were analysed from four cancer patients who had received 50 mg/m2 orally as three divided doses every 8 h. A typical urinary profile of the drug and its metabolites was: parent drug, 13%; 4 demethoxydaunorubicinol (4-DMDNOL), 80%; 4-DMDNR 7-hydroxyaglycone, 4% and 4 DMDNOL 7-hydroxyaglycone, 3%. 4-DMDNOL was the major metabolite detected in plasma. A further metabolite identified as the 7-deoxyaglycone of 4-DMDNOL was detected in plasma of two patients at concentrations equal to or greater than the parent drug. In the other two patients no trace of the metabolite was detected. PMID- 3036388 TI - In vitro evaluation of a new nitrosourea, TCNU, against human small cell lung cancer cell lines. AB - The cytotoxic activity of a new nitrosourea, TCNU, was compared with that of BCNU in five human small cell lung cancer cell lines in vitro. TCNU was found to be equivalent or inferior to BCNU when compared on a microgram to microgram basis. If the potential of in vitro phase II trials for selection of new drugs can be validated, it can be concluded that TCNU is not superior to other nitrosoureas for the treatment of SCCL. PMID- 3036390 TI - Induction of the aromatic hydrocarbon receptor by trans-4-acetylaminostilbene in rat liver. Comparison with other aromatic amines. AB - The effect of the aromatic amines 2-acetylaminofluorene (AAF), 2 acetylaminophenanthrene (AAP) and trans-4-acetylaminostilbene (AAS) on the rat hepatic aromatic hydrocarbon (Ah) receptor level was studied. 3 Methylcholanthrene (MC), as a known receptor ligand, was used as a control. The complete liver carcinogen AAF and MC did not alter significantly the hepatic receptor concentration. In contrast, the strong liver tumor initiator AAS doubled the hepatic Ah receptor level when a dose of 20 mumol/kg was administered for 5 days. AAP increased the amount of the receptor 1.5-fold. PMID- 3036386 TI - Stem cells and the development of mammary cancers in experimental rats and in humans. AB - Evidence based on immunocytochemical staining and ultrastructure suggests that morphological gradations between epithelial and myoepithelial cells, and possibly between epithelial cells and alveolar-like cells occur in terminal ductal structures of rat and human mammary glands. The benign carcinogen-induced rat and benign human mammary tumors can contain epithelial, myoepithelial-like and alveolar-like cells, whereas the malignant counterparts mainly contain only epithelial-like cells. Clonal epithelial cell lines from normal rat mammary glands, benign tumors, and SV40-transformed human mammary glands can differentiate to either myoepithelial-like or alveolar-like cells. In those of the rat, the differentiation processes occur in steps: intermediate cells along the myoepithelial-like pathway resemble intermediates in terminal ductal structures in vivo, and can also generate certain well-differentiated mesenchymal elements of the original tumours. Differentiation of the benign rat cells to alveolar-like cells with mammatrophic hormones and retinoids in vitro leads to a reduction in their tumor-forming ability in vivo. Cell lines from malignant rat mammary tumors of increasing metastatic potential and from human ductal carcinomas largely fail to yield myoepithelial-like or alveolar-like cells and are relatively slow-growing. Growth of the rat mammary epithelial cells in culture is stimulated by a pituitary-derived mammatrophic growth factor (PMGF), prostaglandin E2, and alpha-transforming growth factor; the response of the malignant cell lines to PMGF is reduced. It is suggested that stem cells exist in the rat and human glands that are capable of differentiating to the other major cell types of the mammary parenchyma, and that during the carcinogenic process they generate genetically unstable cells which lose their ability to differentiate and attempt to maximise their intrinsically slow growth rate. PMID- 3036389 TI - Pharmacokinetics of unchanged carboplatin (CBDCA) in patients with small cell lung carcinoma. AB - The disposition of the cisplatin analogue carboplatin was studied in seven patients with small cell lung cancer. Carboplatin 100 mg/m2 was administered without hydration by a 1-h infusion with VP16-213 120 mg/m2 on days 1, 2 and 3 of each course. Plasma and urine collections were made on days 1 and 3 of the first course of treatment. Carboplatin levels in plasma ultrafiltrate and urine were quantitated using a specific and sensitive, high-performance liquid chromatographic assay which involved sample clean-up on a Dowex-2 column prior to injection. Estimates of pharmacokinetic parameters determined using either compartmental or non-compartmental methods were comparable. There was no difference between carboplatin pharmacokinetic parameters determined on days 1 and 3 of treatment. The mean (+/- SD) carboplatin half-life determined from plasma data on day 1 was 105 +/- 30.4 min and was not significantly different from that determined using urinary excretion rate data (107 +/- 51.7 min). Urinary excretion rate plots showed that carboplatin elimination was mono exponential for up to 14 h after infusion. Total-body clearance was 105 +/- 40.0 ml min-1 m-2, renal clearance 64.3 +/- 44.1 ml min-1 m-2, and volume of distribution 17.3 +/- 4.2 l/m2 on the 1st day of treatment. Of the administered dose, 58.4% +/- 21.2% was recovered in urine over a 24-h period after the start of the infusion. The mean renal clearance of carboplatin was comparable to creatinine clearance. Carboplatin disposition was clearly defined in the patients studied using analytical methodology specific for the unchanged drug. PMID- 3036391 TI - Catecholamine-induced myocardial potassium uptake mediated by beta 1 adrenoceptors and adenylate cyclase activation in the pig. AB - The myocardial potassium uptake during intracoronary isoproterenol stimulation was characterized in 12 anesthetized pigs. The beta-receptor subtype specificity and the effect of adenylate cyclase activation were determined. Potassium concentrations were continuously recorded by PVC-valinomycin minielectrodes in the left atrial cavity and in coronary sinus blood diverted through a shunt to the right atrium. The difference in potassium concentration between the left atrial cavity and coronary sinus, and the accumulated myocardial potassium uptake were calculated after computerized data sampling. By intracoronary drug infusion, changes in heart rate and systemic effects were minimized. Isoproterenol (0.6-0.8 microgram/min), a nonspecific beta-agonist, reduced coronary sinus potassium concentration transiently to a nadir of 0.28 (0.15-0.43) mM (median and 95% confidence interval) below control values (n = 12). The potassium uptake, which amounted to 140 (79-202) mumol/100 g tissue, corresponding to an intracellular potassium increase of about 3 mM, was abolished after selective beta 1-blockade by pafenolol. The specific beta 1-agonist dobutamine (40 micrograms/min) caused a similar potassium uptake before and after selective beta 2-blockade by ICI 118, 551. Salbutamol (2 micrograms/min), a specific beta 2-agonist, induced a minor potassium uptake of 4 (1-20) mumol/100 g, blocked by pafenolol. After nonselective beta-blockade by propranolol the adenylate cyclase stimulator forskolin caused a myocardial potassium uptake of similar magnitude to that of isoproterenol before beta-blockade. We conclude that a myocardial potassium uptake ensues during beta 1-adrenoceptor stimulation and adenylate cyclase activation. PMID- 3036392 TI - Quantitative autoradiographic delineation of the distribution of beta-adrenergic receptors in canine and feline left ventricular myocardium. AB - The distribution of adrenergic receptors in specific components of the heart such as vessels and myocytes cannot be determined easily with assays of membranes prepared from homogenates of whole tissue. Accordingly, we characterized the binding of the potent nonsubtype selective antagonist [125iodo]cyanopindolol to beta-receptors in unfixed transmural slices of feline and canine left ventricle. Specific binding ratios greater than 90% were achieved at radioligand concentrations near Kd and greater than 80% at saturating ligand concentrations. Binding of radioligand to receptors in transmural slices was rapid, saturable, stereoselective, and displaceable by antagonists and agonists with the rank order of potency expected of beta-adrenergic receptors. Analysis of binding isotherms indicated maximum binding capacities of 27.8 +/- 6.6 and 40.6 +/- 5.1 fmol/mg tissue protein and dissociation constants of 10.1 +/- 1.8 and 21.3 +/- 1.6 pM in feline and canine ventricular slices, respectively. The distribution of beta receptors in myocytes and selected vascular components of the heart was determined with quantitative film autoradiography and high resolution computer based analysis and display of the density of binding sites, maximum binding capacity, and binding affinity measurements. The results of autoradiographic analysis revealed a uniform transmural distribution of receptors in regions composed primarily of ventricular myocytes but an inverse relation between the density of beta-receptors and the diameter of coronary vessels. Large epicardial conductance arteries had half the receptor density of subjacent myocytes; small mural arteries had approximately 60% of the beta-receptor density of nearby myocytes, and the coronary resistance arterioles had the highest receptor density of any vascular compartment, which was equivalent to that of myocytes. The methods developed should be of particular value in characterizing the distribution and function of receptor subtypes and mechanisms of regulation of adrenergic responsiveness in intact myocardium. PMID- 3036393 TI - Cardiac calmodulin-stimulated protein phosphatase: purification and identification of specific sarcolemmal substrates. AB - A calmodulin-stimulated protein phosphatase has been purified from bovine myocardium. The purification procedure involves sequential DEAE-Sephacel ion exchange chromatography, calmodulin-Sepharose affinity chromatography, and high performance liquid chromatography using a Spherogel TSK DEAE 5PW column. By SDS polyacrylamide gel electrophoresis, the purified cardiac phosphatase consists of two subunits of Mr 61,000 and 19,000, similar to the brain enzyme, calcineurin. Protein phosphatase activity of the cardiac enzyme is stimulated by Ca2+ calmodulin and inhibited by the calmodulin antagonist drug, calmidazolium. Effects of a series of divalent cations on catalytic activity of the cardiac calmodulin-stimulated protein phosphatase are similar to those observed with calcineurin, when the two enzymes are assayed under identical conditions. Highly enriched preparations of bovine cardiac sarcolemma contain substrates of cAMP dependent protein kinase of Mr 166 K, 133 K, 108 K, 79 K, 39 K, and 14 K, which are specifically dephosphorylated by the calmodulin-stimulated phosphatase with pseudofirst-order rate constants of 0.23, 0.46, 0.69, 0.35, 0.69, and 0.115 min 1, respectively. These substrates are not present in purified preparations of cardiac sarcoplasmic reticulum. These results support a role of the calmodulin stimulated phosphatase in the Ca2+-regulation of specific sarcolemmal processes by protein dephosphorylation. PMID- 3036394 TI - Release of adenosine and cyclic AMP from coronary endothelium in isolated guinea pig hearts: relation to coronary flow. AB - The coronary efflux of radioactive 3',5'-cyclic adenosine monophosphate (cAMP) and adenosine from isolated guinea pig hearts was measured following selective prelabelling of coronary endothelial adenine nucleotides with 10 nM [2,8,5'-3H] adenosine. Intracoronary infusion of adenosine and its derivatives 5'-N-ethyl carboxamide-adenosine (NECA), (-)-N6-(R-phenyl-isopropyl)-adenosine (R-PIA), and (+)-N6-(S-phenyl-isopropyl)-adenosine (S-PIA) caused dose-dependent parallel increases in both coronary flow and the coronary efflux of radioactive cAMP with a rank order of potency: NECA greater than R-PIA greater than adenosine greater than S-PIA. In contrast, adenosine receptor stimulation of isolated cardiomyocytes in primary culture decreased the cellular release of cAMP below control levels with a rank order of potency: R-PIA greater than NECA. Under control conditions, coronary efflux of adenosine and cAMP was 34.3 +/- 2.3 and 3.9 +/- 0.8 pmol/min (mean +/- SEM, n = 6), respectively. NECA (12 microM) caused an increase in cardiac cAMP release of 3.8 times and elevated the specific radioactivity of cAMP 5 times to 63.7 +/- 6.0 Ci/mol, a value 11 times greater than the specific radioactivity of tissue ATP. Based on these findings, it was concluded that the coronary endothelium possesses adenosine A2 receptors linked to adenylate cyclase, which are activated in parallel with increases in coronary flow and that cardiomyocyte adenosine receptors are predominantly of the A1 subtype. In addition, the contribution of the coronary endothelium to total cardiac adenosine release was calculated to be 14% using the specific radioactivities of adenosine and cAMP released into the effluent perfusate. PMID- 3036395 TI - Association of decreased myocardial beta-receptors and chronotropic response to isoproterenol and exercise in pigs following chronic dynamic exercise. AB - The effects of chronic dynamic exercise on myocardial beta-adrenergic and muscarinic cholinergic receptors and chronotropic sensitivity to isoproterenol were studied in 5 Yucatan miniswine. Right atrial and left ventricular biopsies, heart rate responses to isoproterenol, and maximal exercise treadmill testing were obtained before and after 10-19 weeks of treadmill running. Radioligand studies using 125I-iodocyanopindolol (ICYP) and 3H-quinuclidinyl benzilate (QNB) were used to determine the number of beta-adrenergic and muscarinic cholinergic receptors. Maximal oxygen consumption increased from 52 +/- 5 to 65 +/- 7 ml/kg/min (mean +/- SD; p less than 0.02), maximal workload from 530 +/- 111 to 1,074 +/- 179 KPM/min (p less than 0.01), resting heart rate decreased from 91 +/ 13 to 62 +/- 4 beats/min (p less than 0.01), heart rate at 75% of pretraining maximal workload decreased from 253 +/- 15 to 196 +/- 12 beats/min (p less than 0.01), and maximal exercise heart rate decreased from 273 +/- 6 to 254 +/- 9 beats/min (p less than 0.01). Decreased heart rate responsiveness to adrenergic stimulation was observed following chronic exercise. Maximal isoproterenol stimulated heart rate decreased from 225 +/- 13 to 185 +/- 28 beats/min (p less than 0.05) and the slope of the isoproterenol dose-response relation decreased from 63 +/- 16 to 40 +/- 16 (p less than 0.05). Radioligand studies revealed a decrease in beta-receptor number in the right atrium following chronic exercise (61 +/- 9 vs. 34 +/- 8 fmol/mg; p less than 0.02), but receptor number in membranes from the left ventricle did not change (60 +/- 9 vs. 62 +/- 4 fmol/mg).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3036396 TI - Characteristics of adrenoceptors and [3H]nitrendipine receptors of porcine vascular smooth muscle: differences between coronary artery and aorta. AB - Characteristics of the bindings of [3H](-)dihydroalprenolol, [125I]( )iodocyanopindolol, [3H]prazosin, [3H]yohimbine, and [3H]nitrendipine to porcine coronary membranes were investigated and the results compared with studies of porcine aortic membranes. In the equilibrium binding study carried out in sarcolemma-enriched fractions, there were no major differences in the Kd values of these radioligands between coronary artery and aorta. However, the densities of beta-, alpha 1-, and alpha 2-adrenoceptors and [3H]nitrendipine receptors of coronary artery were 258, 12, 12, and 561 fmol/mg protein, respectively, while those of aorta were 37, 525, 1,000, and 215 fmol/mg protein. beta-Adrenergic agonists competed with [3H](-)dihydroalprenolol binding sites in coronary artery, the order of potency being (-)isoproterenol greater than (-)norepinephrine greater than (-)epinephrine greater than (+)isoproterenol. In case of aorta, the order was (-)isoproterenol greater than (-)epinephrine greater than ( )norepinephrine. The competition by (+/-)bisoprolol (beta 1-selective antagonist) and ICI 118,551 (beta 2-selective antagonist) for [125I](-)iodocyanopindolol binding sites in coronary artery resulted in nonlinear Hofstee plots (beta 1:beta 2 = 90%:10%). In case of aorta, linear Hofstee plots were obtained. From these results, we conclude that coronary beta-receptors in pigs are predominantly of beta 1-type, while those of aorta are of beta 2-type; regarding the relative population of adrenoceptors, coronary artery is beta-dominant (beta/alpha = 11), while aorta is alpha-dominant (beta/alpha = 0.02); compared with alpha adrenoceptors, coronary artery has a greater number of [3H]nitrendipine binding sites (nitrendipine/alpha-adrenoceptor = 23) than aorta (nitrendipine/alpha adrenoceptor = 0.14). PMID- 3036397 TI - Sudden cardiac death. Neural-cardiac interactions. PMID- 3036399 TI - [Primary approach to visualize the courses of channels by use of isotopes]. PMID- 3036400 TI - Relationship between thyroid hormones and angiotensin-converting enzyme. PMID- 3036398 TI - [The role of alpha and beta receptors and their effects on 5-HT metabolism in acupuncture analgesia]. PMID- 3036401 TI - Initial assessment of tumor-associated antigen CA-125 in patients with ovarian, cervical, and testicular tumors. AB - Serum CA-125, a glycoprotein antigen, has been measured in serum of patients with ovarian cancer, cervical cancer, or nonseminomatous germ-cell tumor of testis. Increased concentrations were found in 21 of 27 patients with epithelial ovarian cancer and two of three patients with ovarian teratoma. Changes in CA-125 concentrations in serum during chemotherapy mirrored the progress of the disease as assessed by clinical and radiological evidence. Although CA-125 provides no real asset for diagnosis, it should have value as a marker for monitoring response to chemotherapy. PMID- 3036402 TI - Automated determination of angiotensin-converting enzyme in serum. AB - This is an adaptation of the Fujirebio "ACEcolor" kit for automated measurement of angiotensin-converting enzyme (EC 3.4.15.1) in serum with the Cobas Fara centrifugal analyzer. The linear range extends to an activity of 110 U/L. Results obtained by the present method and by the manual method were identical, and correlated closely (r = 0.983) with those by Cushman's modified method. The reference interval for 77 adult blood-bank donors was 9-25 U/L (mean 17, SD 4 U/L). Within-run and between-run CVs are 1.7 and 4.0%, respectively. The present method permits rapid, precise, and economical measurement of the enzyme and allows users of a Cobas Fara centrifugal analyzer to introduce a fully automated assay for angiotensin-converting enzyme into their clinical laboratory. PMID- 3036403 TI - Serum angiotensin converting enzyme activity in ex-smokers. AB - A low activity of angiotensin converting enzyme (ACE) has been reported in people who smoke. To determine whether this low ACE activity would be reversible on cessation of smoking, we measured serum ACE activity in 107 healthy male volunteers. They included 27 active cigarette smokers, 28 non-smokers, 24 ex smokers who had stopped smoking for less than 10 yr, and 28 ex-smokers who had stopped smoking for more than 10 yr. The mean value (+/-SD) of serum ACE in those who had stopped smoking for more than 10 yr was comparable to that of non smokers: 23.2 +/- 5.1 and 23.5 +/- 4.5, respectively. ACE activity in smokers and the ex-smokers who had stopped smoking for less than 10 yr was significantly lower (17.8 +/- 4.5 and 17.8 +/- 3.9, respectively) than values obtained in non smokers and the group who had not smoked for more than 10 yr (p less than 0.001). These findings suggest that the effect of chronic smoking on the serum ACE activity may be reversible. PMID- 3036404 TI - 5'-Nucleotide phosphodiesterase isoenzymes in human serum: quantitative measurement and some biochemical properties. AB - A method based on native PAGE is used for the quantitative measurement of 5' nucleotide phosphodiesterase isoenzymes (5'-NPD; EC 3.1.4.1) in human serum. In contrast to other techniques this method works with a commercially available substrate. In sera of healthy donors four isozymes could be separated, designated as 5'-NPD-I, 5'-NPD-II, 5'-NPD-III and 5'-NPD-IV in the reverse order of their electrophoretic mobility. When low amounts of serum were applied to the gel, the separation between all four activities was sufficient enough to allow their quantitation. Higher amounts of serum impaired the separation between the isoenzymes I and II. However, even when using high amounts of serum, the quantitation of these activities was possible when taking advantage of some of their biochemical properties which are described herein. PMID- 3036405 TI - A rapid and precise assay for calmodulin and cyclic AMP phosphodiesterase on a centrifugal analyser. AB - An assay for cyclic AMP phosphodiesterase in biological samples is described. The method is a continuous, spectrophotometric assay adapted for use on a centrifugal analyser. Using the calmodulin-activated cyclic AMP phosphodiesterase prepared from pig brain by DEAE-cellulose chromatography, and a standard calmodulin preparation from the same source, a rapid assay for calmodulin is described. The calmodulin assay has a working range of 2 to 6 mg/l calmodulin in the sample. The within-batch CV is less than 5% and the between-batch CV is 7.0% or less. PMID- 3036406 TI - In vitro effects of thymosin and lithium on lymphoproliferative responses of normal donors and HIV seropositive male homosexuals with AIDS-related complex. AB - The in vitro effects of thymosin fraction 5 (TF5) and lithium chloride (LiCl) on the ability of peripheral blood mononuclear cells (PBMC) obtained from 37 normal male donors and 33 male patients with AIDS-related complex (ARC) to respond to alloantigenic stimulation (mixed leukocyte reaction, MLR) and to produce interleukin 2 (IL-2) in response to mitogens were studied. TF5 significantly increased MLR responses in normal donors (P less than 0.01) and in a group of 33 ARC patients with depressed cellular immunity (P less than 0.05). Similar effects were observed when LiCl was added to the MLR assays in both the normal and the ARC patient groups. Furthermore, TF5 and LiCl exhibited additive immunoenhancing properties. In 10 normal donors TF5 enhanced phytohemaggutinin (PHA)-induced IL-2 production as well as IL-2 production in response to pokeweed mitogen (PWM) (P less than 0.02). TF5-mediated enhancement of IL-2 production by PBMC obtained from ARC patients was observed in response to both mitogens, i.e., PHA and PWM. Additionally, LiCl increased PHA-induced IL-2 production in both normal subjects and ARC patients. LiCl and TF5 together had an additive effect in the enhancement of IL-2 production in both groups of subjects. Our data extend previous observations regarding the immunoregulatory activities of TF5 and LiCl and provide evidence that PBMC obtained from ARC patients have the potential to respond in vitro to these agents. The significance of these findings is discussed. PMID- 3036407 TI - Antihypertensive and renal effects of enalapril in post-transplant hypertension. AB - The acute and chronic antihypertensive and renal effects of the angiotensin converting enzyme inhibitor, enalapril, were studied prospectively in ten hypertensive renal transplant recipients. Acute administration of enalapril produced a significant decrement in both systolic and diastolic blood pressure but had no significant effect on glomerular filtration rate or effective renal plasma flow. The antihypertensive effect of enalapril was enhanced by gradually increasing the dose of the drug or by addition of a diuretic during six to eight weeks of chronic therapy. During chronic enalapril therapy, four patients developed renal insufficiency that reversed after discontinuation of the drug. In three of these four cases, overt renal insufficiency was associated temporally with the addition of a diuretic. Digital angiography revealed unequivocal transplant renal-artery stenosis in three of the four patients with renal insufficiency; the fourth patient had diffuse narrowing of the transplant renal artery without a discrete stenosis. It is concluded that enalapril alone or in combination with a diuretic is effective in lowering blood pressure in patients with post-transplant hypertension. The development of renal insufficiency during enalapril therapy may be exacerbated by concomitant diuretic therapy and should raise the suspicion of underlying transplant renal-artery stenosis. The acute blood pressure or renal response to a small dose of enalapril does not reliably predict the development of renal insufficiency during treatment with larger doses of the drug. PMID- 3036408 TI - Behcet's syndrome, a personal view. PMID- 3036409 TI - Asymmetric involvement of the spinal cord involving both large and small anterior horn cells in a case of familial amyotrophic lateral sclerosis. AB - A case of familial amyotrophic lateral sclerosis revealing neuronal loss in the anterior horns and Clarke's columns, Lewy body-like intracytoplasmic inclusions in some remaining cells, cord-like thickening of cell processes, mild perivascular infiltration of lymphocytes, and degeneration of the middle root zone in the posterior column and of the spinocerebellar tract is described. Two highly unusual features were noted. First, there was a marked asymmetry of the neuronal loss and tract degeneration in the spinal cord. The left side was much more severely affected than the right side. Second, both large and small neurons disappeared almost completely on the left side of the lumbar anterior horn. PMID- 3036410 TI - Intraosseous glomus tumor in the ulna. A case report. AB - Glomus tumors are benign vascular tumors composed of round to oval uniform cells associated with vascular structures, usually found in the soft tissue of the nail bend. A 24-year-old woman with a two-year history of progressive right elbow pain and swelling was found to have a histologically characteristic intraosseous glomus tumor in this unusual location. Excision was followed by complete relief of symptoms. PMID- 3036411 TI - Rapid diagnosis of herpes encephalitis by enzyme immuno-assay. AB - Five cases of encephalitis caused by herpes simplex virus (HSV) are described. HSV-specific IgM and IgM antibodies were detected in cerebrospinal fluid and serum by use of antibody-capture noncompetitive enzyme-linked immunosorbent assay. The tests seem suitable for rapid diagnosis in the second week after onset of neurological symptoms. The clinical importance of early diagnosis and therapy in patients of HSV encephalitis has been discussed. PMID- 3036412 TI - Ileal leiomyosarcoma and leiomyoma. False-positive scintiscans for Meckel's diverticulum. AB - Two cases of ileal leiomyomatous neoplasms with positive scintigraphic findings for Meckel's diverticulum are presented. The precise mechanism for the abnormal concentration of the Tc-99m pertechnetate is uncertain. Illustrations of the collating barium and computerized tomographic studies are included and the scanning technique utilized is reviewed. PMID- 3036413 TI - Receptor redistribution does not accompany terbutaline-induced down regulation of beta-adrenergic receptors on human mononuclear leukocytes. AB - Mononuclear leukocytes are easily accessible cells for investigating the regulation of beta-adrenergic receptors in humans. We have previously shown that brief incubations with agonists redistribute (? internalize) most of the beta adrenergic receptors on mononuclear leukocytes away from the cell surface without changing total receptor number. However, negligible redistribution occurred after exercise or an infusion of isoproterenol. The current study was designed to ask whether receptor redistribution occurs over a longer time course after administration of terbutaline, a beta 2-adrenergic agonist that is known to cause a decrease in receptor number. Normal volunteers were given terbutaline, 5 mg t.i.d. for 6 days. As expected, the number of beta-adrenergic receptors decreased. However, the remaining receptors were not redistributed. Redistribution also did not occur after 1 or 2 days of terbutaline, at which time down regulation was minimal. We also found that terbutaline did not alter the ability of the receptors to be redistributed or desensitized by a preincubation with isoproterenol. PMID- 3036415 TI - Influence of cyclo-oxygenase inhibition and of leukotriene receptor blockade on pulmonary vascular pressure/cardiac index relationships in hyperoxic and in hypoxic dogs. AB - Overall mean pulmonary arterial pressure (MPAP)/cardiac index (CI) relationships were investigated in 13 pentobarbital anaesthetized dogs ventilated consecutively with a fraction of inspired O2 (F1O2) of 0.4 and with a F1O2 of 0.1. This sequence of alternated F1O2 0.4 and F1O2 0.1 was repeated in the dogs with a strong pulmonary pressor response to hypoxia (more than 20% increase in pulmonary vascular resistance) (n = 6) under a continuous infusion of the leukotriene receptor blocker FPL 57231 (2 mg min-1 kg-1), and in the dogs with a weak pressor response to hypoxia (n = 7) after cyclo-oxygenase inhibition by acetylsalicylic acid (1 g intravenously). Five-point MPAP/CI plots were constructed by opening a femoral arteriovenous fistula or by stepwise inflations of an inferior vena cava balloon catheter. The MPAP/CI plots were rectilinear in all experimental conditions. In responders, hypoxia was associated with an increase in MPAP over the entire range of CI studied (1-5 litres min-1 m-2). Infusion of FLP 57231 abolished the vasoconstricting effect of hypoxia. In non-responders, MPAP was not affected by hypoxia over the entire range of CI. After acetylsalicylic acid administration, hypoxia resulted in a significant rise in MPAP from 2 to 5 litres min-1 m-2. Infusion of FLP 57231 decreased mean systemic arterial pressure at both F1O2 0.4 and F1O2 0.1, while acetylsalicylic acid had no effect on systemic haemodynamics.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3036416 TI - [Role of endogenous opioid peptides in the pathogenesis of circulatory shock]. PMID- 3036414 TI - Effects of verapamil and converting enzyme inhibition on bilateral renal function of two-kidney, one-clip hypertensive rats. AB - Experiments were conducted in two-kidney, one-clip renal vascular hypertensive rats (GHR) to assess the responses of each kidney to acute treatment with the antihypertensive calcium channel blocking agent verapamil in the presence and in the absence of converting enzyme inhibitor (CEI). One group of GHR (0.2 mm inner diam. clip 3 weeks before study) were examined during a control period, and during a second period of infusion of verapamil (600 micrograms h-1 kg-1). A second group of GHR were examined during a control period, during CEI (teprotide, 3 mg h-1 kg-1) infusion and during a third period of verapamil (600 micrograms h 1 kg-1) infusion superimposed on CEI infusion. Although systemic blood pressure (BP) decreased from 175 +/- 4 to 149 +/- 5 mmHg (mean +/- SEM) in response to verapamil alone, renal blood flow for non-clipped kidneys increased from 5.9 +/- 0.4 to 6.5 +/- 0.3 ml/min, indicating a 30% reduction of renal vascular resistance (P values less than or equal to 0.01; n = 9). Glomerular filtration rate (GFR) for non-clipped kidneys (n = 24) increased from 0.91 +/- 0.09 to 1.47 +/- 0.14 ml/min and filtration fraction increased from 0.32 +/- 0.04 to 0.47 +/- 0.03 (P values less than or equal to 0.05). Urine flow rate and absolute and fractional sodium excretion for non-clipped kidneys increased. GFR for clipped kidneys decreased during verapamil. Treatment with CEI alone resulted in nearly identical responses of BP and function of the non-clipped kidney, except filtration fraction was unchanged. The addition of verapamil to ongoing converting enzyme blockade tended to augment the increased GFR of the non-clipped kidney. Plasma renin activity (PRA) increased from 30 +/- 3 to 59 +/- 7 ng of angiotensin (ANG) I h-1 ml-1 with verapamil alone, a significantly larger increment than the increase of PRA from 27 +/- 5 to 39 +/- 9 ng of ANG I h-1 ml-1 in GHR subjected to comparable blood pressure reduction by mechanical aortic constriction. Verapamil resulted in many similar effects on renal function to those observed during blockade of converting enzyme. The increased filtration fraction observed in response to verapamil may be the result of vasodilatation of the afferent arteriole or of an increase in the glomerular ultrafiltration coefficient. PMID- 3036417 TI - Metabolism of glycerate 2,3-P2--XI. Essential amino acids of pig phosphoglycerate mutase isozymes. AB - Phosphoglycerate mutase isozymes (types M, B and MB) from pig tissues are inactivated upon treatment with reagents specific for histidyl, arginyl and lysyl residues. Their mutase, 2,3-bisphosphoglycerate synthase and 2,3 bisphosphoglycerate phosphatase activities are concurrently lost, although some differences exist in the rate of inactivation. No significant differences are observed between the isozymes. The reversion of the modifying reactions reactivates the three enzymatic activities. Substrates and cofactors protect against inactivation, the protective effects varying with the modifying reagent. Titration with pCMB shows the existence of two essential thiol groups per subunit type M. These results provide evidence of the intrinsic character of the three enzymatic activities, favor their location at the same active site and suggest the existence of separate binding sites for monophosphoglycerates and bisphophoglycerates. Both type M and B subunit from pig phosphoglycerate mutase are similar to type M subunit from rabbit and to the enzyme from yeast. PMID- 3036418 TI - Activity of ATP synthetase complex after low temperature treatment or freeze drying of mitochondria isolated from skeletal muscles. AB - The influence of freezing, thawing, or freeze-drying on ATP synthetase complex of isolated skeletal muscle mitochondria was studied. Cooling to -60 or to -196 degrees C and rapid thawing did not change activity significantly. Slow warming stimulated the release of latent ATP-ase activity and decreased ATP synthesis. These changes were more pronounced after freeze-drying. PMID- 3036419 TI - Calcium- and phospholipid-dependent protein kinase C and phosphatidylinositol kinase: two major phosphorylation systems in the cornea. AB - The signaling messengers for the regulation of intracellular calcium-dependent functions involve a phospholipid-sensitive protein kinase (protein kinase C) modulated by diacylglycerol, which is a product of phosphoinositide degradation. We found that protein kinase C activity is two times higher in the epithelium than in the stroma-endothelial layers of the rabbit cornea. 12-0 tetradecanoylphorbol-13-acetate (TPA) and phosphatidylserine stimulate protein kinase C at low Ca2+ concentrations. The TPA-phosphatidylserine stimulated activity of corneal epithelium was recovered in the soluble fraction; in the membrane-bound fraction, a very active phosphatidylinositol kinase accounted for half the basal phosphorylation of the corneal epithelium. PMID- 3036420 TI - Sodium and potassium saturation kinetics of Na+K+-ATPase in plasma membranes from corneal endothelium: fresh tissue vs. tissue culture. AB - The maximal velocity (Vmax) and the apparent dissociation constant (K0.5) of Na+K+-ATPase have each been estimated with respect to sodium and potassium ion activation. These estimations were made from the enzymatic activity of plasma membrane preparations derived from bovine corneal endothelial cells. The determinations were made on cells obtained from fresh tissue and from secondary tissue cultures. Two methods were used to obtain the estimates: the first used a combination of Eadie-Hofstee and Hill plots; the second used Eisenthal-Cornish Bowden plots. The Vmax for sodium was 5.58-5.60 mumol Pi/mg protein/30 min for fresh tissue vs. 2.00-1.80 mumoles Pi/mg protein/30 min for tissue cultures. The corresponding K0.5 values were 62-57 mM (fresh tissue) vs. 7.9-6.7 mM (tissue culture). Vmax for potassium was 4.28-4.00 mumoles Pi/mg protein/30 min for fresh tissue vs. 1.37-1.34 mumoles Pi/mg protein/30 min for tissue cultures. The corresponding K0.5 values were 3.3-3.1 mM (fresh tissue) and 1.7-1.7 mM (tissue culture). The results indicate a lowered activity and change in affinity of the enzyme for the two ions in tissue cultures compared to fresh tissues. The detergent Lubrol W-X increased activity in both tissue sources. Sonication had no significant effect on the activity. Variations in pH (7-9) indicated that the highest activity was obtained at pH 7.8 for the enzyme in tissue culture while activity was highest at pH 8.0 for the enzyme in fresh tissues. These differences in kinetic activity suggest a response to changes in the ion requirements of these cells due to their environment, developmental stage or some other parameter. PMID- 3036421 TI - Colchicine as a probe for aberrant collagenase expression in recessive junctional epidermolysis bullosa. PMID- 3036422 TI - Quantitation of hepatitis B virus DNA in serum of patients with hepatoma. AB - Hepatitis B virus (HBV) DNA in the serum of 31 patients with histologically confirmed primary hepatocellular carcinoma (PHC) from Malaysia and Indonesia was quantitated by densitometric scanning of autoradiograms obtained by Southern blot DNA hybridization, after electrophoresis using a 32P DNA cloned into plasmid pBR325 as a probe. This quantitation after electrophoresis is more informative than the usual spot hybridization technique. Five of the 31 sera were positive for HBV DNA. Levels ranged between 1.36 pq and 143.18 pq per ml of serum, and the levels of HBsAg, anti-HBs, anti-HBc, HBeAg and anti-HBe in the serum were serologically determined. All five sera positive for HBV DNA were also positive for HBsAg. Three of the five positive for HBV DNA were positive for HBeAg and negative for anti-HBe. Two of the sera positive for HBV DNA were negative for HBeAg but positive for anti-HBe. All sera negative for HBV DNA were also negative for HBeAg. Many sera which were negative for HBV DNA and HBeAg were positive for HBsAg. Of the 31 sera from PHC patients, 23 had at least one HBV marker positive (74.2%). PMID- 3036423 TI - The human neurofilament gene (NEFL) is located on the short arm of chromosome 8. AB - We have localized the gene coding for the human neurofilament light chain (NEFL) to chromosome band 8p2.1 by Southern blotting of DNA from hybrid cell panels and in situ hybridization to metaphase chromosomes. PMID- 3036424 TI - Extracellular matrix injury during lung inflammation. PMID- 3036425 TI - Pulmonary artery obstruction due to malignant fibrous histiocytoma. AB - Obstruction of the pulmonary artery can be due to a variety of conditions, including chronic thromboembolic disease, neoplasia, and fibrosing mediastinitis. Differentiation can be difficult. We report a case of malignant fibrous histiocytoma causing intravascular obstruction and include a review of the literature. PMID- 3036426 TI - Isoenzyme pattern of enolase and creatine kinase in small cell lung cancer patients. PMID- 3036427 TI - Proton magnetic resonance imaging to stage activity of interstitial lung disease. AB - Patients with active interstitial lung disease (ILD) are often treated with high dose corticosteroids, although such therapy is not universally effective and has significant risks. Clinicians have utilized various ancillary diagnostic techniques to help with difficult treatment decisions. Since magnetic resonance imaging (MRI) has the theoretic and experimental potential of differentiating various stages of ILD, we prospectively studied 34 adult patients in a 0.15 Tesla resistive magnet body imager. The most severely affected patients, who were thought to warrant a steroid trial, had the greatest MRI image intensity, and dramatic improvement was seen following successful treatment. Qualitative MRI data were as useful as any other ancillary diagnostic technique (radiographic, physiologic, scintigraphic) in predicting clinical course. Computer-generated relaxation times were not sufficiently precise to differentiate active from inactive disease. Although limited by availability and cost, MRI appears to be a useful ancillary diagnostic technique in ILD patients facing immunomodulating therapy. PMID- 3036430 TI - Secondary hematologic changes in small cell lung cancer and malignant lymphoma. PMID- 3036428 TI - Transbronchial needle aspiration in the diagnosis of bronchogenic carcinoma. AB - Transbronchial needle aspiration (TBNA) was performed as a diagnostic procedure in 91 consecutive patients ultimately proven to have bronchogenic carcinoma. Results of TBNA were compared, in the same patients, to the diagnostic yield of cytologic examination of sputum, endobronchial brushings and washings, and endobronchial/transbronchial biopsy. The diagnostic yield for sputum was 13 percent (10 of 75); brushings, 40 percent (34 of 84); washings, 29 percent (26 of 89); biopsy, 56 percent (42 of 75); and TBNA, 45 percent (41 of 91). Aspirates were positive in 35 percent of patients with adenocarcinoma, 41 percent with squamous cell carcinoma, 52 percent with large cell undifferentiated carcinoma, and 55 percent of patients with small cell carcinoma. Carinal aspirates were positive in 54 percent (6 of 11); paratracheal aspirates, 57 percent (13 of 23); parabronchial aspirates, 39 percent (11 of 28); endobronchial, 78 percent (7 of 9), and peripheral mass or solitary pulmonary nodule, 40 percent (17 of 42). The overall diagnostic yield for brushings, washings, and biopsy was 64 percent. The addition of TBNA increased the yield to 71 percent. Bronchogenic carcinoma was diagnosed solely by TBNA in six patients, all with extrabronchial or extratracheal lesions. We conclude that TBNA increases the diagnostic yield of bronchoscopy, particularly in patients with extratracheal and extrabronchial lesions. An equally important observation is that TBNA fails to contribute significantly to the diagnosis of cancer in patients with lesions readily accessible by conventional bronchoscopic techniques. Exceptions to this observation include occasional patients with necrotic endobronchial tumors, submucosal lesions, and rarely patients with peripheral lung nodules or masses. PMID- 3036429 TI - Roles of some inflammatory mediators in the pathogenesis of early pulmonary edema in steam respiratory burns. PMID- 3036431 TI - Differentiation of poliovirus type I isolates in the 1982 epidemic from Sabin-I strain by antigenic and temperature markers. AB - Poliovirus type 1 strains recovered from patients with poliomyelitis during the epidemic of 1982, were distinct antigenically from Sabin-1 strain by comparing homologous and heterologous neutralization rate constants of Sabin-1 specific antiserum. With the reproductive capacity at 40 degrees C, these isolates could be easily differentiated from vaccine strain. It was suggested that the epidemic of poliomyelitis in 1982 was not caused by vaccine strain. The 24 isolates reacted with LSc and/or Sabin-1 antisera to the same extent. It was speculated that they might be antigenically similar if not identical. Three isolates which were collected from the later part of epidemic formed small plaques at 40 degrees C. Rabbit antisera against poliovirus strains 3844, Sabin-1, Mahoney and Brunhilde were produced. Antigenic relatedness among the strains was tested by kinetic neutralization. The strain 3844 was distinct from Brunhilde, Mahoney and Sabin-1. Asymmetric cross-reactions between Mahoney and Sabin-1 strains were clearly demonstrated. PMID- 3036433 TI - [Combined reconstruction of the bile ducts and liver vessels in cancer of the hepatic duct at its bifurcation]. PMID- 3036432 TI - [A rare case of polydactyly with syndactyly and a supernumerary metatarsus]. PMID- 3036434 TI - The histopathologic examination of the effects of hydrochloric acid-pumice abrasion bleaching technique on the pulp in rat teeth. PMID- 3036435 TI - [Immunity and immunogenetics of trophoblastic neoplasms]. PMID- 3036436 TI - Localization polymorphism of EBV DNA genomes in the chromosomes of Burkitt lymphoma cell lines. AB - The localization of Epstein-Barr virus (EBV) genomes in nuclei of the human lymphoblastoid cell lines Raji, Jijoye, P3HR-1, Daudi and Ramos was investigated by in situ hybridization with biotinylated EBV DNA probes. We found that all sites of hybridization were associated with the chromosomes. Only some of these sites were present on both chromatids and these had a non-random distribution; these sites could represent EBV sequences integrated at specific points on the chromosomes. The total mean site number corresponded with the number of viral DNA copies estimated in the different cell lines by other techniques, but the copy number was highly variable from cell to cell in a given line. PMID- 3036437 TI - [Diagnostic significance of the combined determination of serum alpha-1 antitrypsin and alpha-1-antichymotrypsin for primary hepatocellular carcinoma]. PMID- 3036438 TI - Small round viruses: classification and role in food-borne infections. AB - Since the first observation of Norwalk virus in the electron microscope in 1972, many different small virus particles in the size range 20-40 nm have been described world-wide in association with outbreaks of gastroenteritis. Progress characterizing these agents has been hampered by the relatively small numbers of particles present in clinical material and the lack of success in culturing them. Although the relationship between some of these viruses remains confusing, a number of distinct groups has emerged, based on morphological features and limited physical data. Immuno-electron microscopy has proved valuable in detecting viruses but the addition of antibody can mask surface morphological features. Examination of viruses in negatively stained preparations without added antibody has revealed distinct morphological differences and viruses previously thought to be simply antigenic variants within the Norwalk group of viruses clearly belong to other groups. Preliminary evidence suggests that one human virus unrelated to Norwalk has a single-stranded DNA genome and is a parvovirus. Some groups have been implicated in outbreaks of food-borne gastroenteritis, particularly after the consumption of shellfish, and their role in other food borne and water-borne outbreaks is being increasingly recognized. PMID- 3036439 TI - The candidate caliciviruses. AB - The Caliciviridae are a family of small (35-40 nm) RNA viruses with a characteristic cupped morphology. They are unique in possessing only a single major structural polypeptide, of Mr 60,000-71,000. The use of electron microscopy to investigate diarrhoeal diseases has revealed viral particles with the size and structure of the caliciviruses in the faeces of humans, domestic and farm animals, birds, reptiles and insects. In vivo experiments indicate that they are species specific and have confirmed that they replicate in the gut, which often results in the host developing diarrhoea and failing to thrive. Biochemical characterization of these agents has been hampered by a failure to produce sufficient yields of virus in vitro. However, fluorescence and radiolabelling experiments indicate that the human, canine and chicken viruses replicate in the cytoplasm and possess an RNA genome. A major structural polypeptide (Mr 60,000 71,000) has been identified in the human, canine and insect viruses. Diagnosis of the candidate caliciviruses is dependent on electron microscopy and fluorescence labelling, with the exception of the human agents, for which radioimmunoassays have been developed. There is little epidemiological information on these agents but there is increasing evidence that the human caliciviruses are a common cause of outbreaks of diarrhoea and vomiting in infants, adults and the elderly. PMID- 3036441 TI - Comparative pathology of infection by novel diarrhoea viruses. AB - Examination of diarrhoeic faeces in the electron microscope often reveals viruses that are presumed to be enteropathogenic. Lesions caused by novel rotaviruses were similar to those of group A rotaviruses, but enterocyte syncytia were seen which are probably pathognomonic for novel rotaviruses. In adenovirus infection in piglets, mature enterocytes were infected and destroyed; intranuclear inclusion bodies were seen in infected enterocytes. Calici-like viruses infected mature enterocytes in calves and the lesions were similar to those described in humans infected with calici-like viruses; in both host species it was impossible to demonstrate virus particles in enterocytes examined in the electron microscope. The Breda virus infected villi and crypts in the lower small intestine and the surface and crypts in the large intestine; it was the only enteropathogenic virus to show this distribution of infection and lesions. Astrovirus infection in lambs was comparable to a mild rotavirus infection, but in calves the epithelium of the dome villi of Peyer's patches was infected. Parvovirus in cats and dogs infected and destroyed small intestinal crypt cells, causing dilated crypts and stunted villi; intranuclear inclusion bodies were prominent. PMID- 3036442 TI - Clinical trials of rotavirus vaccines. AB - The clinical efficacy of candidate rotavirus vaccines has been tested in Tampere, Finland, over four winter and spring rotavirus epidemic seasons in 1983-1986. Testing against natural challenge has demonstrated that heterologous oral rotavirus vaccines induce cross-protection to human rotavirus diarrhoea. The trials have also given insight into mechanisms of protection in human rotavirus diarrhoea. After the oral vaccination of infants aged six to 12 months the highly attenuated bovine rotavirus strain RIT 4237, titre 10(8) per dose, probably 'takes' in most vaccinees, producing a symptomless intestinal infection with a low virus excretion rate and an antibody response in over 80% of the initially seronegative subjects. Upon natural challenge such vaccination gives no protection against human rotavirus infection but gives 50-60% protection against any clinically detectable rotavirus-associated illness and 80-90% protection against severe rotavirus diarrhoea, regardless of the infecting human rotavirus serotype. The less attenuated rhesus monkey rotavirus RRV-1, titre 10(5)-10(6) per dose, is more infectious in humans, and virus multiplication in the intestine results in excretion of vaccine virus in the stools and some clinical symptoms, mainly fever, 3-4 days after vaccination. The degree of protection against human rotavirus diarrhoea appears similar to that induced by bovine rotavirus vaccine. PMID- 3036440 TI - Immunobiology of Norwalk virus. AB - Clinical immunity to Norwalk virus in inoculated human volunteers appears to be unusual for gastroenteritis viruses, as certain individuals are repeatedly ill on long-term virus rechallenge and others remain persistently well. In these volunteers there is a paradoxical inverse correlation between the prechallenge serum (and jejunal fluid) Norwalk antibody level (measured by radioimmunoassay) and resistance to illness, suggesting that non-immunological factors, perhaps genetic, may be important in determining resistance. Most reported naturally occurring Norwalk disease outbreaks in developed nations also show that humoral antibody fails to correlate with immunity to infection. The unusual pattern of clinical immunity to Norwalk virus indicates a need for caution in the development of vaccines against this agent as well as a need for additional information on its immunobiological characteristics. The virus is known to contain a single protein, like the caliciviruses. Recently we have found evidence for at least a one-way serological cross-relatedness between Norwalk virus and human calicivirus. Twelve of 20 paired sera from ill patients in outbreaks due to calicivirus strain UK4 seroconverted to Norwalk virus by radioimmunoassay and two of eight paired sera from UK2 outbreaks showed seroconversion. Future studies of outbreaks caused by various calicivirus strains should be designed to correlate acute-phase serum antibody titres to Norwalk virus with clinical susceptibility and immunity to infection. PMID- 3036443 TI - The diagnostic gap in diarrhoeal aetiology. AB - It is well established that rotaviruses of group A are the most important cause of severe diarrhoea in children. The causes of most cases of infectious diarrhoea still remain unidentified, however, and there must be other viruses to be found. 'Novel' rotaviruses have recently been discovered, mainly in animals (serogroups B and C in pigs and humans, D in birds, and one or more further groups in sheep and other mammals). Except for the group B virus which has caused widespread outbreaks of quite severe diarrhoea in adults in China (still not reported from outside China) these novel rotaviruses are rarities in the human and probably represent uncommon zoonotic infections. We speculate that the Chinese virus might have arisen by reassortment of genetic segments of animal group B viruses or perhaps by mutation, and so became infectious for man. The problem of identifying and determining the importance of small round viruses is reviewed. It seems likely that the group of small, round structured viruses, including Norwalk and the viruses of plainly calicivirus morphology, are all representatives of a whole group of enteric caliciviruses. Until they can be more easily cultivated it will be difficult to make diagnostic reagents available to all. Astroviruses and genuine parvoviruses have been found by many people in many countries. Astroviruses are probably more important as pathogens in lambs than in children or calves; parvoviruses can only be established as significant in epidemics. The coronavirus-like particles, first found in Vellore and Bristol, are still enigmatic and their role in diarrhoea is uncertain. Toroviridae, recently discovered as causes of diarrhoea in ungulates, do not seem to be at all important as causes of diarrhoea in humans. Possibly fruitful approaches to future searches are outlined: firstly to make more extensive use of immuno electron microscopy; and secondly to try to improve existing tissue culture systems to make them more sensitive to enteric viruses. PMID- 3036444 TI - Nucleic acid-based analyses of non-group A rotaviruses. AB - Simple genome profile studies on polyacrylamide gels allow all non-group A rotaviruses isolated so far to be recognized by the absence of the tight triplet (7-9) of RNA segments seen in all group A viruses. However, reliance solely on genome profile studies for rotavirus grouping can be misleading and, for virus group definition, additional corroborating nucleic acid and serological studies are essential. Terminal fingerprint analysis was the first generation of nucleic acid-based assays that allowed discrimination between the various rotavirus groups. By means of this technique the clear definition of five rotavirus groups (A-E), correlating exactly with those found by a serological assay, has been possible, with preliminary evidence for at least two additional groups. The technical sophistication of fingerprinting techniques prevents their widespread use in epidemiological studies; the development of a second generation of nucleic acid-based assays is therefore under way. These employ molecularly cloned cDNA probes to the genomes of non-group A viruses which can be widely distributed for use in 'dot-blot' screening of faecal samples and, if expressed as protein in Escherichia coli, should provide a ready source of viral antigen for use in surveying viral prevalence through the screening of serum antibody levels. PMID- 3036446 TI - Novel rotaviruses in animals and man. AB - Novel (non-group A) rotaviruses have many of the morphological, biochemical and biological properties described originally for group A rotaviruses but they do not share the same group antigens. By negative-stain electron microscopy, novel rotaviruses have the characteristic rotavirus morphology, although with some novel rotaviruses the characteristic single- and double-shelled particles may not be readily apparent. Comparison of novel rotaviruses in serological tests has revealed the existence of at least six rotavirus serogroups, A to F, with the original rotaviruses belonging to group A. As with group A rotaviruses, viruses from different animal species, including man, can belong to the same serogroup. A further point of difference between novel and group A rotaviruses is their genome profiles, which lack the triplet of segments in the 7-8-9 region of group A rotaviruses. This is a useful diagnostic aid. Novel rotaviruses have been found in farm animals and man. They can cause enteritis experimentally and infect villus enterocytes. In chickens, turkeys, lambs and pigs the viruses and/or antibody to them are commonly found, in association with either clinical or subclinical infection. In humans one type of novel virus has emerged as a cause of severe diarrhoeal disease in adults. The possible reasons for the relatively recent discovery of the novel rotaviruses are discussed. PMID- 3036445 TI - Seroepidemiology and molecular epidemiology of the Chinese rotavirus. AB - The Chinese rotavirus which causes epidemics of diarrhoea in adult humans was isolated in 1983. This virus, designated adult diarrhoea rotavirus (ADRV), resembles typical rotaviruses morphologically and has a genome made up of 11 discrete segments of double-stranded RNA. Because the Chinese rotavirus has a unique RNA pattern on polyacrylamide gel electrophoresis and is antigenically distinct from group A rotaviruses, it is tentatively included in group B. Infection with ADRV or ADRV-related viruses (as shown by serological study) is detected in human populations as widespread as mainland China, Hong Kong, Australia, the United States and Canada, and in some domestic animals. RNA co electrophoresis has shown homology of isolates from 12 different outbreaks (with some minor variations at segments 10, 11, 3 and 5). cDNA probes and monoclonal antibodies have been prepared to improve the detection and further characterization of the virus. PMID- 3036447 TI - Enteric adenoviruses. AB - The 41 serotypes of human adenoviruses are classified into six subgenera (A-F) with different tropisms. Enteric infections are caused in children by serotypes Ad40 and Ad41 of subgenus F. Serotypes Ad40 and Ad41 transform embryonic cells but cannot induce tumours in newborn hamsters. They differ from all other (established) human adenoviruses by being unable to replicate in conventional cell cultures. Ad40 and Ad41 grow in 293 cells (human embryonic kidney cells immortalized by transfection with the E1A, E1B regions of Ad5). In spite of the difficulty of isolating Ad40 and Ad41 they can be directly identified in stools by enzyme-linked immunosorbent assay (ELISA) and solid-phase immuno-electron microscopy. The amount of viral DNA in stool preparations is sufficient for identification by DNA restriction or dot-blot analysis. Adenoviruses have been associated with 7-17% of cases of diarrhoea in children. Ad40 and Ad41 cause diarrhoea throughout the year. Clinical features are watery stools, vomiting and moderately elevated temperature; respiratory symptoms are infrequent. The diarrhoea is protracted (mean 8.6 and 12.2 days for Ad40 and Ad41 respectively). Children with rotavirus diarrhoea vomited more frequently and had a higher temperature and diarrhoea of shorter duration. The impact of enteric adenoviruses in the aetiology of diarrhoea world-wide is not known but is accessible to investigation. PMID- 3036448 TI - Metabolic changes during the defunctionalized stage after ileal pouch-anal anastomosis. AB - Metabolic changes between the preoperative period and the defunctionalized stage after ileal pouch-anal anastomosis were investigated in 21 patients. Aspects studied included weight change, renal function, liver function, lipid metabolism, and two hematologic parameters. Of 21 patients, 19 lost weight. Cholesterol levels decreased from 183 +/- 30 IU/1 to 122 +/- 48 IU/1 (P less than .0001). Triglyceride levels rose from 95 +/- 29 IU/1 to 190 +/- 86 IU/1 (P less than .01). Significant elevations were seen in values of four liver function tests: SGOT, SGPT, lactic dehydrogenase, and alkaline phosphatase (P less than .05). Blood urea nitrogen levels increased from 11.1 +/- 4.7 mg/100 ml to 21.8 +/- 24.8 mg/100 ml (P less than .05). Serum creatinine levels rose from 0.97 +/- .18 mg/100 ml to 1.5 +/- 1.2 mg/100 ml (P less than .05). Uric acid levels increased from 5.6 +/- 1.8 mg/100 ml to 7.5 +/- 2.3 mg/100 ml (P less than .001). Hemoglobin values did not change significantly. Platelet counts rose from 304,000 +/- 79,000 to 447,800 +/- 189,000 (P less than .05). Identification of these systematic alterations in metabolic parameters during the defunctionalized stage can aid the physician in management of these patients. PMID- 3036449 TI - Factitious hypoglycemia mimicking insulinoma. PMID- 3036450 TI - Retroperitoneal lymph node aspiration biopsy in staging of pelvic cancer: a cytological study of 228 consecutive cases. AB - Knowledge of the status of the pelvic lymph nodes is important for accurate staging and adequate treatment of patients with genitourinary tract cancer (bladder and prostatic carcinoma, testicular tumors, and uterine carcinoma). A total of 228 consecutive patients underwent preoperative evaluation of the lymph node status by lymphangiography combined with fine-needle aspiration biopsy. A lymphadenectomy was performed in 94 patients. The overall diagnostic accuracy was 93%. There were 5% false-negative results and no false-positive diagnoses. Fine needle aspiration biopsy is an inexpensive method with which to detect the presence of metastatic lymph nodes visualized by bipedal lymphangiography. It is also a safe and well-tolerated method, with a low morbidity and no mortality. The various cytological features are described as are some possible pitfalls. PMID- 3036451 TI - Fine-needle aspiration biopsy of abdominal and retroperitoneal tumors in infants and children. AB - Fine-needle aspiration biopsy has been employed to establish morphologic diagnoses in abdominal and retroperitoneal tumors in 54 infants and children. A 0.4-0.7-mm gauge needle was used; the puncture was performed through the anterior abdominal wall in the abdominal tumors and by the lateral approach in the retroperitoneal tumors. Malignant tumors were discovered in 51 cases (94.4%). The nature of the lesion was recognized in 96.2%, and correct cell typing was achieved in 90.2%. The most frequent lesion was non-Hodgkin's malignant lymphoma, followed by neuroblastoma, nephroblastoma, and individual cases of other epithelial and mesenchymal tumors. Using parallel bone marrow examination, half of the neuroblastoma cases and five of 28 lymphoma cases showed bone marrow involvement. We observed no complications caused by the fine-needle aspiration technique. Due to the safety and efficacy of this technique, it can often be used instead of explorative laparatomy. PMID- 3036452 TI - Fine-needle aspiration cytology of terminal duct carcinoma of minor salivary gland. AB - The cytologic features of terminal duct carcinoma of the palate, as observed in a fine-needle aspiration specimen, are described and contrasted with the cytologic features reported for benign mixed tumor, basal-cell adenoma, and adenoid cystic carcinoma. Terminal duct carcinoma, at times, may be difficult, if not impossible, to distinguish from adenoid cystic carcinoma in fine-needle aspiration specimens. In most instances, this distinction may not be important. PMID- 3036453 TI - Cyclic AMP regulates transcription of the genes encoding human chorionic gonadotropin with different kinetics. AB - Choriocarcinoma cell lines characteristically synthesize and secrete the alpha- and beta-subunits of human chorionic gonadotropin (hCG), as well as the intact heterodimer. Treatment of one such cell line, BeWo, with 8-bromo-adenosine 3':5' cyclic monophosphate (8-Br-cAMP) causes at least a 10-fold increase in the concentration of the mRNA encoding each subunit. Changes in mRNA concentrations are associated with similar changes in transcription rates of both the CG alpha and CG beta genes, although the kinetics of their transcriptional responses are different. Transcription of the alpha-subunit gene increases rapidly and becomes maximal within 1 hr after addition of 8-Br-cAMP. By contrast, transcription of the CG beta gene increases slowly and progressively for at least 8 hr after treatment with 8-Br-cAMP. The slow transcriptional response of the CG beta gene(s) appears to be unique compared to that of other cAMP-responsive genes, and suggests that the cyclic nucleotide may regulate transcription of the CG genes by different mechanisms. PMID- 3036455 TI - [Evaluation of irregular hepatectomy for primary liver carcinoma]. AB - From 1964 to 1985, 120 patients with primary liver carcinoma were treated by operation in our hospital. Regular hepatectomy was done in 7 patients, palliative irregular hepatectomy in 28 and radical irregular in 85. The operation mortality was 4.2% in irregular hepatectomy group (113 cases) but 14.3% in regular hepatectomy group (7 cases) (P greater than 0.05). The 1, 3 and 5 year survival rates were 68.8%, 48.1% and 20.0% in radical irregular hepatectomy group but 83.3%, 33.3% and 16.7% in regular hepatectomy group. 10 of 28 patients treated by palliative hepatectomy were added with radiation. Majority of these patients died in 1 year after operation but 2 patients survived for more than 2 years and 1 for more than 7 years. The data show that in Asia, the incidence of primary liver carcinoma concurrent with liver cirrhosis is high and irregular hepatectomy is a suitable treatment. There is no difference between irregular and regular hepatectomy groups in the prognosis. But the former could reduce the operative time, mortality and the possibility of bleeding and complications. PMID- 3036454 TI - Recombination of selectable marker DNA in Nicotiana tabacum. AB - A chimeric neomycin phosphotransferase II (NPT II) gene, which normally provides kanamycin resistance to transformed plant cells, was inactivated by in vitro deletions. Repair plasmids not containing plant-specific transcription signals but containing only the NPT II coding region (or parts of it) were used in co transformation experiments involving direct DNA uptake into protoplasts isolated from Nicotiana tabacum. Recombination, or gene conversion mediated by homologous sequences produced active NPT II genes in about 1% of transformants, rendering these cells resistant to kanamycin. Analysis of the size of the active enzyme indicated that recombination had occurred producing an NPT II gene indistinguishable from the wild-type gene. Southern blot analysis revealed that the bulk of co-transformed donor plasmid DNA had suffered structural modifications; however, kanamycin resistance was inherited in a Mendelian fashion, indicating that at least one functional and structurally intact copy of the regenerated NPT II gene is integrated into the host genome. PMID- 3036456 TI - [Retrospective study on cancer of the base of the tongue]. AB - From Feb. 1962 to Feb. 1982, 54 patients with cancer of the base of tongue were treated in our hospital. There were 44 squamous cell carcinomas and 10 adeno cystic carcinomas. 46 patients were treated by radiotherapy only, 6 by surgery plus radiotherapy, 1 by surgery and 1 by chemotherapy alone. The 5 year survival rate was 27.1%. The result indicates that radiotherapy alone is poor. The author suggests that for cancer of the base of tongue, a combination of radiotherapy and surgery be used and prospective randomized study be carried out. The defect can be repaired using skin or myocutaneous flaps. In extensive lesions, glossectomy plus laryngectomy should be considered. For the healthy and young patients, the lesions of the base of tongue together with a part of the larynx should be extirpated but for the older man, selective glossectomy is advised. PMID- 3036457 TI - [Acute reversible kidney insufficiency due to enalapril during diuretic-treated heart insufficiency]. AB - Acute reversible renal failure with hyperkalemia developed in a 42-year-old woman during treatment of heart failure and hypertension with high doses of enalapril and diuretics. Creatinine clearance fell as low as 9 ml/min. Renal-artery stenosis and prerenal renal failure were excluded. Reduction in blood pressure in conjunction with reduced renal perfusion in heart failure can make glomerular filtration angiotensin dependent. Interruption of angiotensin II formation in this situation triggers off functional renal insufficiency. PMID- 3036458 TI - [Glycoside uptake in the myocardium]. PMID- 3036460 TI - An elucidation of the arachidonic acid cascade. Discovery of prostaglandins, thromboxane and leukotrienes. AB - Arachidonic acid is normally stored in membrane-bound phospholipids and released by the action of phospholipases. Enzymatic conversion of released arachidonic acid into biologically active derivatives proceeds through one of several routes. Cyclo-oxygenase converts arachidonic acid to unstable cyclic endoperoxides from which prostaglandins, prostacyclin and thromboxanes are derived. Formation of the leukotrienes from arachidonic acid is initiated by the action of 5-lipoxygenase producing leukotriene A4. Hydrolysis of leukotriene A4, or the incorporation of glutathione results in the formation of leukotriene B4 and C4, respectively. In addition, 12- and 15-lipoxygenase can catalyse arachidonic acid conversion and lipoxins A and B are amongst the possible products. Many of these metabolites of arachidonic acid feature prominently in the development of inflammation. Prostaglandin E2 and prostacyclin are potent vasodilators, while leukotriene D4 causes cellular adhesion, chemotaxis of neutrophils and degranulation. Leukotrienes C4, D4 and E4 contribute to inflammation by increasing vascular permeability. Leukotrienes are also believed to play an important pathophysiological role in allergic broncho-constriction of asthma. Through pharmacological intervention in the arachidonic acid cascade various anti inflammatory agents have been developed. These include aspirin-like drugs, which inhibit cyclo-oxygenase. Corticosteroids appear to indirectly inhibit phospholipases thus preventing release of arachidonic acid. Future progress in this field is likely to produce drugs which antagonise arachidonic acid derivatives or inhibit the enzymes involved in their synthesis with greater specificity. PMID- 3036459 TI - The role of arachidonic acid metabolites in local and systemic inflammatory processes. AB - Leukotrienes are synthesised from arachidonic acid via the 5-lipoxygenase pathway in neutrophils, eosinophils, monocytes/macrophages, basophils and certain mast cell populations. Their synthesis is closely regulated by several known factors and the cells which contain 5-lipoxygenase do not all possess the capability to synthesise all of the leukotrienes. Neutrophils produce leukotriene B4, which attracts other neutrophils, whereas the leukotriene C4, produced by eosinophils, increases the contractile activity of smooth muscle. Monocytes/macrophages are able to produce both of these leukotrienes. Receptor sites for leukotriene B4 have been identified on monocytes and neutrophils and receptors for leukotriene D4, a cleavage product of leukotriene C4, have been defined in pulmonary tissue. In animals, sulphidopeptide leukotrienes have been shown to cause potent vasoconstriction resulting in increased blood pressure and increased vascular permeability leading to hypovolaemia. These leukotrienes also depress renal (in animals) and pulmonary (in animals and humans) function, the latter probably as a result of effects on peripheral rather than central airways. In patients with mild asthma, however, there is no differential activity of this type. The sulphidopeptide leukotrienes caused wheal and flare when administered intradermally in healthy volunteers, which was of considerably longer duration than that induced by prostaglandin D2. Conversely, leukotriene B4 caused accumulation of neutrophils in the absence of wheal and flare. Studies into the effects of dietary fish oil showed that 2 constituents, docosahexanoic acid and eicosapentaenoic acid (EPA), inhibit the conversion of arachidonic acid by cyclo oxygenase, but not by 5-lipoxygenase. Furthermore, 5-lipoxygenase converts EPA to a pentene series of leukotrienes and the sulphidopeptide derivatives possess similar activity to their tetrameric counterparts.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3036461 TI - Pathogenesis of inflammation. Effects of the pharmacological manipulation of arachidonic acid metabolism on the cytological response to inflammatory stimuli. AB - It is generally accepted that non-steroidal anti-inflammatory drugs (NSAIDs) act by the inhibition of prostaglandin biosynthesis. However, this hypothesis may not apply to all NSAIDs at all doses. It has thus been proposed that, in high doses, NSAIDs counteract inflammation by the inhibition of the activity of inflammatory cells such as the neutrophil. Neutrophils are activated by a 'twin signal' which is a part of a general stimulus response coupling mechanism involving phospholipid remodelling. This 'twin signal' consists of the mobilisation of intracellular calcium and the activation of protein kinase C. NSAIDs inhibit the aggregation of neutrophils in vivo and in vitro, and show additive effects with stable prostaglandins on the inhibition of the generation of superoxide anion, a product of inflammation. Binding studies have shown that the mechanism of this effect appears to be an interference with the ligand-receptor site modulated by the G protein. Activation of the cells of the marine sponge, which are not responsive to stable prostaglandins and do not contain a cyclo-oxygenase, can be inhibited by NSAIDs. Therefore, further investigation of this interesting hypothesis should help to clarify the precise mechanism of the anti-inflammatory effects of these drugs. PMID- 3036462 TI - Three ATPase activities have an abnormal developmental time course in trembler sciatic nerves. AB - Trembler mutant mice are affected by a peripheral neuropathy characterized by hypomyelination, demyelination, and Schwann cell proliferation. In adult mutants, supernumerary Schwann cells form membranous structures known as 'onion-bulb' formations. The activities of the Na+, K+-ATPase and of two ouabain-insensitive Mg2+-ATPases were investigated in sciatic nerves of young and adult mutants. The Na+, K+-ATPase activities were 92 and 76% of the control values in young and adult mutants, respectively. By immunoblot analysis, the alpha-subunit of the Na+, K+-ATPase had an identical apparent molecular weight in controls at both ages and in young mutants. In adult mutants, on the contrary, the alpha-subunit appeared smaller by about 2 kd, similar to that in kidney, indicating that the Na+, K+-ATPase was localized mainly on supernumerary Schwann cells. In addition, in the mutants, the developmental increase of both the mitochondrial and the nonmitochondrial Mg2+-ATPase was abnormally high. We suggest that the abnormal increase of the nonmitochondrial Mg2+-ATPase activity during development reflects an enrichment of that enzyme in 'onion-bulb' formations. PMID- 3036464 TI - Histopathological changes in liver and thyroid of the teleost fish, Channa punctatus (Bloch), in response to ammonium sulfate fertilizer treatment. AB - In Channa punctatus exposed to safe (100 ppm) and sublethal (500 ppm) concentrations of the commonly used fertilizer ammonium sulfate for 6 months, from January to June, hepatocytes revealed initial hypertrophy followed by exhaustion as evidenced by degranulation, nuclear pyknosis, and focal necrosis. Thyroid follicles exhibited various degrees of hypertrophy, hyperplasia, hyperemia, and reduction in colloid content. Both in the liver and in the thyroid, changes were more pronounced in the 500-ppm-treated fish. These results suggest that this fertilizer, which is washed into the water system in small quantities, may cause dose-dependent dysfunction of liver and thyroid. PMID- 3036463 TI - Iron deficiency in ethnic minorities: associations with dietary fibre and phytate. AB - Routine full blood counts and serum ferritin determinations were carried out after admission to hospital in 112 children which included a white caucasian group (n = 65) and two ethnic minority groups of West Indians (n = 24) and Asians (n = 23). In these 3 groups those children between the ages of 1 week and 6 months were found to have similar haemoglobin, mean corpuscular volume, and ferritin levels. In the remaining children (aged from 7 months to 14 years 5 months) serum ferritin levels were lower in the ethnic minority groups than in white caucasians, but the haemoglobin and mean corpuscular volume were not significantly different. Children with lower height centiles had reduced ferritin levels, irrespective of their ethnic origins. A nutritional survey between the ages of 7 months and 14 years 5 months showed that mean daily dietary intakes of energy, protein, and iron in white caucasions were similar to those in West Indian and Asian children. The differences noted were in larger phytate and fibre intakes in the ethnic minority groups. Asian diets appeared to differ in containing meat less often as a source of iron, while pulses and chapattis provided more phytate and fibre. It is suggested that dietary intakes of phytate and fibre are important in causing lower ferritin levels by reducing iron absorption. PMID- 3036465 TI - [Purple membranes as a model system for research on photoreception processes. III. The bacteriorhodopsin photocycle and proton transport]. PMID- 3036467 TI - The role of subunit sialic acid in the thyrotropic and gonadotropic activities of human chorionic gonadotropin. AB - The present studies were undertaken to delineate the role of sialic acid residues in the two subunits of hCG in relation to its hormonal activity. Sialic acid was removed by treatment of the individual subunits or intact hCG with insolubilized neuraminidase. Desialylated variants of hCG were obtained by combining an asialo subunit (as alpha or as beta) with its complementary intact or desialylated subunit. When tested in the hCG receptor assay using a rat testis fraction, none of the hCG variants exhibited any loss of activity compared with that of intact purified hCG. In in vitro bioassays that employed cAMP and testosterone generation in rat Leydig cells as indices of response, intact hCG and as alpha in combination with intact beta (as alpha-beta) were equipotent. In contrast, intact alpha-subunit combined with as beta (alpha-as beta) and desialylated intact hCG (asialo-hCG) showed activity that at the highest concentration of each tested (20 ng/ml) was no more than half of that evoked by intact hCG. In a TSH receptor assay in human thyroid membranes, loss of sialic acid from either one or both hCG subunits resulted in enhancement of binding affinity; the rank order of decreasing potency was asialo-hCG, alpha-as beta, as alpha-beta. However, despite their enhanced binding affinity, and like intact hCG itself, none of the variants elicited a cAMP response in either human thyroid membranes or cultured human thyroid cells. Rather, asialo-hCG and alpha-as beta, but not intact hCG and as alpha-beta, were effective antagonists of the stimulatory response induced by bovine TSH in thyroid cells. These findings indicate that desialylation of hCG enhances its binding affinity for hCG receptors in testis and, much more so, for TSH receptors in human thyroid. Desialylation also changes hCG from a full agonist to a partial agonist in testis and from a nonagonist to an antagonist in human thyroid. In all cases, sialic acid in the beta-subunit of hCG appears to have a predominant role in these effects. PMID- 3036468 TI - Regulation of cholesterol side-chain cleavage enzyme activity by gonadotropin in rat corpus luteum. AB - The locus of gonadotropin-induced acute stimulation of pregnenolone production by cholesterol side-chain cleavage (CSCC) enzyme containing cytochrome P450 (cytP450scc) was examined in rat corpus luteum. Mitochondria were isolated from pseudopregnant rat ovaries after treatment with different doses of human CG (hCG) (25-200 IU) for 30 min; Electron Paramagnetic Resonance (EPR) spectra of high spin cholesterol complex of cyt P450scc (type I high spin EPR signal) and the cyt P450scc activity were determined. hCG treatment increased the formation of type I EPR spectra compared to that obtained with saline-treated controls, and pretreatment with cycloheximide (30 mg/kg BW) before hCG abolished this increase. The magnitude of type I EPR signal diminished with increasing pH over the range of 6.2-7.3. The type I EPR signal increased with doses of hCG and correlated well with the pregnenolone production. Aminoglutethimide treatment (competitive inhibitor of CSCC) before hCG injection led to an increased accumulation of cholesterol in inner mitochondrial membranes with a corresponding decrease in the outer membrane cholesterol, and cycloheximide treatment inhibited this accumulation. This suggests that the transport of cholesterol to inner mitochondrial membranes from outer membranes is regulated by hCG. In addition, gonadotropin also regulates the redistribution of cholesterol within the inner mitochondrial membranes. PMID- 3036466 TI - The inhibition of low density lipoprotein metabolism by transforming growth factor-beta mediates its effects on steroidogenesis in bovine adrenocortical cells in vitro. AB - Transforming growth factor-beta (TGF beta) has a differential effect on the growth and function of bovine adrenocortical cells in vitro. TGF beta inhibits basal as well as ACTH- or angiotensin II-stimulated steroid formation, with no evidence of change in cell growth. The major inhibitory effect of TGF beta occurs at a step before cholesterol formation, since treatment of adrenocortical cells with TGF beta decreased not only delta 4-steroid levels but also delta 5-steroid levels. The addition of cholesterol reverses the suppression of steroidogenesis induced by TGF beta. To determine the mechanism of this inhibition, the effect of TGF beta on low density lipoprotein (LDL) metabolism was investigated. Cells treated with TGF beta showed a significant suppression of [125I]iodohuman LDL ([125I]LDL) binding to the cell surface, followed by decreases in internalization and proteolytic degradation of [125I]LDL. Maximal inhibition of LDL metabolism was observed at a concentration of 1 ng/ml (4 X 10(-11) M) TGF beta. The stimulation of LDL metabolism by ACTH was also inhibited by TGF beta, and the inhibition observed correlated well with the inhibition of steroidogenesis. The inhibitory effect of TGF beta on [125I]LDL binding results from the decrease in the maximal LDL-binding capacity. The stimulation of LDL uptake induced by Bu2cAMP, cholera toxin, forskolin, and Ang II was also decreased by treatment with 1 ng/ml TGF beta. The specificity of this effect is quite high, since the inhibitory effects of TGF beta on LDL metabolism were not observed with either inhibin A or activin, two molecules that have considerable structural homology to TGF beta. We conclude that TGF beta specifically suppresses LDL metabolism in bovine adrenocortical cell cultures and that this step may mediate, at least in part, its role as a potent inhibitor of steroidogenesis. PMID- 3036469 TI - Ovarian adrenergic nerves directly participate in the control of luteinizing hormone-releasing hormone and beta-adrenergic receptors during puberty: a biochemical and autoradiographic study. AB - The rat ovary receives sympathetic innervation from the superior ovarian nerve (SON) and the plexus nerve (OP). To examine the possibility of a direct adrenergic mechanism controlling ovarian receptor distribution during the onset of puberty, we have studied the acute (48-h) effect of unilateral nervotomy (combined section of SON and OP nerves) on ovarian LHRH and beta-adrenergic receptor concentrations and distribution using both radioreceptor assays and in vitro autoradiography. Ovarian LHRH receptor concentration increased sharply between 12 and 20 days of age. At this time receptors were mostly associated with follicles and interstitial cells, whereas at 37 days of age, when a measurable loss in the receptor concentration was observed, light and diffuse autoradiographic labeling of receptors was also found in the corpora lutea. Complete removal of adrenergic input to the gland produced a sharp decrease in LHRH-binding activity within the denervated ovary at each time interval studied, with no effect in the innervated contralateral gland. Autoradiographic data also revealed a decrease in both the number of labeled follicles and the intensity of the labeling. beta-Adrenergic receptor concentration increased progressively between days 12 and 27, reaching a peak value at 37 days of age. Labeling was very weak at 12 days of age and increased progressively at 20 and 27 days of age. At this time, receptors were mostly localized by autoradiography in the interstitial cells, while at 37 days of age corpora lutea were strongly labeled. Ovarian beta-adrenergic receptors showed a marked drop when acutely deprived of their neural tone, as illustrated by the 2- to 3-fold decrease in receptor binding capacity within the denervated gland. The autoradiographic data also showed marked changes in beta-adrenergic receptor distribution, specially at 37 days of age. At this time, the labeling of corpora lutea was markedly decreased in denervated ovaries. The present results clearly demonstrate that complete removal of ovarian adrenergic tone produces a profound decrease in the concentrations of LHRH and beta-adrenergic receptors within the ovary, although it cannot be excluded that peptidergic factors also arriving via the SON and OP could have some influence on the regulation of these receptors. The results support the concept of a direct involvement of the central nervous system in ovarian function. They also suggest that during ovarian development a neural efferent system might be involved in the adjustment of ovarian responsiveness to stimulation by the gonadotropins via changes in receptor content and/or distribution in the different ovarian compartments. PMID- 3036470 TI - The effect of polymyxin B on steroidogenesis from adrenocortical cells. AB - The action of the cyclic peptide polymyxin B (a well known inhibitor of protein kinase C) on adrenal steroid synthesis was examined with Y-1 adrenal tumor cells. Polymyxin B produces a biphasic effect on the stimulation of steroid synthesis by 2 nM ACTH in these cells, with inhibition at low concentrations (less than 10 microM) and a return to control levels at high concentrations (greater than 100 microM). Polymyxin B does not inhibit the stimulatory effect of Bu2cAMP on steroidogenesis. Inhibition of the steroidogenic response to ACTH by a fixed concentration (20 microM) of polymyxin B is overcome by high concentrations of ACTH. Polymyxin B causes a concentration-dependent stimulation of steroid synthesis by Y-1 cells and, over the same concentration range, increases the production of cAMP by these cells. Polymyxin B also partially inhibits the increased production of the cyclic nucleotide produced by ACTH. In addition, polymyxin B inhibits binding of [125I](Phe2,Nle4)ACTH-(1-38) to Y-1 cells. Polymyxin B, like ACTH, promotes rounding of Y-1 cells and partially inhibits rounding produced by ACTH. These effects of polymyxin B are specific to the extent that polymyxins E1 and E2 do not exert similar effects. The actions of polymyxin B are not confined to transformed cells, since responses similar to those seen with Y-1 cells were also observed with cultured rat fasciculata cells. On the other hand, the effect of polymyxin B is specific for adrenal cells, since the cyclic peptide does not influence the steroidogenic response of rat Leydig cells to LH. It is concluded that polymyxin B is a partial agonist of ACTH which is likely to prove useful in studying the molecular basis of the interaction between ACTH and its adrenal receptor. PMID- 3036471 TI - Opiate receptor subtypes in the rat hypothalamus and neurointermediate lobe. AB - The potent opiate radioligands [3H]etorphine, [3H]ethylketocyclazocine (EKC), and [3H]naloxone, bound specifically and saturably to a single class of membrane binding sites in rat neurointermediate lobe (NIL), with Kd values of 3.7, 24, and 51 nM, respectively. In the hypothalamus (Ht), [3H]etorphine bound to specific and saturable sites with a Kd of 2.9 nM. Binding-inhibition studies with [3H]etorphine and unlabeled etorphine-HCl as well as [3H]EKC and unlabeled EKC, revealed high and low affinity binding sites in rat Ht and NIL as well as in the neural lobe of the bovine pituitary gland. [3H]naloxone also bound specifically to two classes of sites in Ht membranes, but to only a single class of low affinity sites in NIL membranes. Specific binding represented 80-90% of total [3H]etorphine binding, about 75% of total [3H]EKC binding, and 45-55% of total [3H]naloxone binding at 22 C in NIL and Ht, respectively. Relative binding potencies derived from Ki values for binding-inhibition studies of [3H]etorphine with opioid peptides and opiates were: NIL, etorphine-HCl greater than dynorphin A greater than naloxone-HCl greater than dynorphin-(1-9) greater than beta endorphin much greater than alpha-neoendorphin approximately (Leu5)enkephalin approximately DAGO (Tyr-D-Ala-Gly-NMe-Phe-Gly-ol); Ht, etorphine HCl greater than naloxone-HCl greater than beta-endorphin greater than dynorphin A much greater than DAGO greater than morphiceptin much greater than (Leu5)enkephalin. Specific [3H]etorphine binding was also demonstrable after preincubation of NIL membranes with DAGO and (Leu5)enkephalin and after preincubation of Ht membranes with morphiceptin and (Leu5)enkephalin; such binding could be displaced by nonradioactive dynorphin A. In addition, [3H]etorphine binding to bovine neural lobe was displaceable by naloxone-HCl, with an ED50 of 43 nM. Specific ligands for sigma-opiate receptors, such as (+)SKF 10,047 (N-allylnorcyclazocine), phencyclidine (PCP), and (-)cyclazocine, displaced specifically bound [3H]etorphine and [3H]EKC from NIL membranes only at high (micromolar) concentrations. However, specific [3H]PCP sites were of higher affinity in NIL and Ht membranes, with similar Kd values of 102 and 190 nM respectively, and different concentrations (0.15 and 1.32 pmol/mg protein, respectively). These data have revealed several differences in the opiate-binding properties of rat Ht and NIL membranes.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3036472 TI - Electrical properties of cultured human adrenocorticotropin-secreting adenoma cells: effects of high K+, corticotropin-releasing factor, and angiotensin II. AB - ACTH-secreting pituitary adenoma cells were cultured from specimens obtained by transphenoidal hypophysectomy in five patients with Cushing's disease. The majority of adenoma cells (90%) stained specifically with antiserum against human ACTH. The electrophysiological properties and response to hormones of these cells were studied with intracellular recording techniques under current clamp and voltage clamp conditions. Most (80%) of the cells fired action potentials that were Ca2+-dependent inasmuch as they were blocked by Co2+ (5 mM) and by removal of Ca2+ from the medium, but were unaffected by tetrodotoxin (0.3 mM) and by Na+ removal. The cells responded to factors known to stimulate ACTH release, including high K+, CRF, and angiotensin II (AII). High K+ (50 mM) induced a membrane depolarization in association with an increase in conductance. CRF (100 nM) produced a depolarization, a decrease in conductance, an increase in spike firing, and an increase in spike duration. Although AII was inactive in ordinary recordings, in cells loaded with lithium (Li+) to promote the phospholipid dependent second messenger system, the peptide produced an increase in spike firing and spike duration with no change in membrane potential. The combination of CRF and AII (CRF + AII; 100 nM each) in Li+-loaded cells caused a greater excitatory effect than either peptide alone. Under voltage clamp, the response either to CRF or to CRF + AII could be attributed, at least in part, to the inhibition of a slow, voltage-dependent K+ current that is persistently active at resting potential. These results indicate that modulation of action potential firing may be an early step in the regulation of ACTH release from pituitary cells by known secretagogues. Since action potentials in these cells are associated with Ca2+ entry, the resulting changes in intracellular Ca2+ levels could mediate the effects of the hormones on secretion. PMID- 3036473 TI - Biological effects of aluminum on normal dogs: studies on the isolated perfused bone. AB - Although it is well known that aluminum (Al) plays a role in the development of osteomalacia in patients with chronic renal failure, the mechanisms are not fully understood. Since the osteoblasts are the cells responsible for the formation of osteoid tissue, which is greatly affected in patients with Al-induced osteomalacia, it is possible that Al could affect the number of osteoblasts or interfere with their function. To further characterize this potential mechanism, we performed studies in isolated perfused tibiae from normal and Al-treated dogs. In this system, when PTH is added to the perfusate, cAMP, a major marker of osteoblasts, is released. The dogs were divided into two groups: control, and Al treated (0.75 mg/kg, iv, 5 days a week for 3 months). Thereafter, the dogs were killed, and the tibiae were perfused in vitro. PTH-(1-34) (3-4 ng/ml) and 3 isobutyl-1-methylxanthine (an inhibitor of phosphodiesterase) were added to the perfusate. Basal cAMP secretion was the same in both groups of dogs. After PTH was added to the perfusate, cAMP increased to a peak of 188.2 +/- 30.6 pmol/min in the normal dogs vs. 113 +/- 8.15 in Al-treated dogs (P less than 0.05). Cumulative cAMP secretion over a 30-min period was 766 +/- 127.9 pmol in the normal dogs vs. 455.6 +/- 38.2 pmol in the experimental animals (P less than 0.05). The histological appearance of bone biopsies taken before and after Al administration are consistent with a suppressive effect of the cation on osteoblast function. In particular, the number of osteoblasts had decreased 8 fold (P less than 0.01) under the influence of Al, and tetracycline-based measurements of mineralization kinetics show that osteoblast-mediated calcification was dysfunctional (P less than 0.01-0.025). On the other hand, the histological features of the post Al treatment biopsies suggest that at some time during its administration, the cation stimulates osteoblastic activity. For example, new (woven) bone formation was present in two dogs, and in another, lamellar bone, deposited under the influence of Al, covered the entire trabecular surface. Moreover, Al-associated osteoid was deposited independent of prior resorptive activity, indicating that the cation promotes bone formation in the absence of prior resorption. In keeping with its trophic effect on matrix deposition, Al also led to extensive marrow fibrosis in five dogs, indicating that Al also stimulates the activity of fibroblasts, cells closely related to osteoblasts.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3036474 TI - Lack of difference between retinoic acid and retinol in stimulating progesterone production by luteinizing granulosa cells in vitro. AB - Receptors for retinoids in the immature rat ovary and the effects of retinol and retinoic acid on luteinizing granulosa cells were studied. Radioreceptor assay demonstrated the presence of specific cellular retinol-binding protein and cellular retinoic acid-binding protein in the ovaries of rats injected with PMSG alone or PMSG and hCG. In addition, when luteinizing granulosa cell from PMSG/hCG injected immature rats were cultured with or without retinoic acid, the morphology, viability, number of cells in culture, and progesterone (P) accumulation were not affected by up to 10 microM retinoic acid. Beyond 10 microM, the cells began to round up, which was associated with a decrease in cell viability. Surprisingly, the deleterious concentrations of retinoic acid increased progesterone accumulation significantly higher than the medium control value. This increase in progesterone, however, was not accompanied by an increase in cAMP. When cells preincubated for 2 days with 1 microM of either retinoic acid or retinol were subsequently incubated in retinoid-free medium containing various substrates for steroidogenesis, the following results were obtained. Basal progesterone and its accumulation in response to human low density lipoprotein were significantly higher in cells preincubated with retinoids than in the control cells. However, no difference was seen in the degree of stimulation between retinol and retinoic acid pretreatments. Both 25-hydroxycholesterol, a substrate for side-chain cleavage enzyme, and pregnenolone, a substrate for 3 beta-hydroxysteroid dehydrogenase, significantly stimulated the accumulation of progesterone in cells preincubated with retinoids over the control value. Again, no appreciable difference was observed between retinol and retinoic acid pretreatments. Our results suggest that receptors for retinoids are present in gonadotropin-primed immature rat ovaries, retinoids increase luteal cell progesterone accumulation, and no difference exists between retinol and retinoic acid in their ability to increase the accumulation of progesterone by these cells. PMID- 3036475 TI - Rhythmicity of triiodothyronine generation by type II thyroxine 5'-deiodinase in rat pineal is mediated by a beta-adrenergic mechanism. AB - Type II T4 5'-deiodinase plays an essential role in converting T4 to T3 in extrathyroidal tissues, thus allowing the full development of a cellular effect of thyroid hormone. This enzyme is one of the most important factors in regulating local action of thyroid hormone. Its activity in the rat pineal was 20 30 times higher at midnight than at noon; this nocturnal rise was abolished by the beta-adrenergic blocker, propranolol. The beta-agonist isoproterenol, caused a 3-fold increase in diurnal enzyme activity. There is thus a marked nyctohemeral variation in T4 activation in the pineal, probably mediated by a beta-adrenergic mechanism. Since the temporal peak of pineal T4 activation corresponds to that of pineal function, our data suggest a previously unrecognized role of thyroid hormone in pineal regulation. PMID- 3036476 TI - Cloning the Fisher rat thyroid cell line (FRTL-5): variability in clonal growth and 3,'5'-cyclic adenosine monophosphate response to thyrotropin. AB - To investigate the stability of FRTL-5 cells we cloned stock cells by limiting dilution in a 6-hormone medium with bovine TSH (bTSH) (10 mU/ml) (6H medium). One third of the wells with irradiated fibroblast feeder cells developed FRTL-5 colonies. One hundred and twenty clones were obtained, of which 29 (8%) developed sufficiently for functional analysis. Clones were tested for growth responses to bTSH stimulation as evidenced by 72-h [3H]thymidine uptake and TSH receptor activation by cAMP generation. Clonal growth responses to bTSH were of three types: dose-related growth increase, absence of growth stimulation, and stimulation by bTSH in concentrations up to 100 microU/ml and inhibition of growth above 100 microU/ml. All clones tested showed evidence of extracellular cAMP accumulation in response to bTSH. However, sensitivity to bTSH varied from 1 to 100 microU/ml and maximum cAMP secretion with 1 mU/ml bTSH varied from 2 to 13 pmol/ml. Whereas certain clones showed high sensitivity to bTSH with respect to both growth and cAMP responses (e.g. 1B-6), there were clones which showed disparity in this relationship, as evidenced by poor growth dependency but high cAMP responses or by growth stimulation yet insensitivity with respect to cAMP secretion. These data demonstrate that the FRTL-5 line contains cells with variable responsiveness to bTSH. Whereas the FRTL-5 line is heterogeneous and subject to developmental variation, cloning by limiting dilution allows the derivation of highly bTSH-sensitive cells, such as 1B-6, from the stock cultures. PMID- 3036477 TI - Hypothyroidism and abnormalities in the kinetics of thyroid hormone metabolism in rats treated chronically with polychlorinated biphenyl and polybrominated biphenyl. AB - Studies were directed at the question of whether polychlorinated biphenyl (PCB; Aroclor 1254) and polybrominated biphenyl (PBB; Fire Master BP-6), when administered in the diets of female Sprague-Dawley rats over long periods of time (5-7 months) and at low dosages (0, 1, 5, 10, and 50 ppm), would depress the thyroid. By examining serum T3 and T4, kinetics of T4 metabolism, and in vivo thyroid response to exogenous TSH injections, an estimate of the degree of hypothyroidism was made, and abnormalities in T4 disappearance from serum were encountered. Serum T3 and T4 levels were greatly suppressed in a dose-related manner by PCB or PBB treatment. There was a diminished response of serum T3 and T4 to TSH injection in rats pretreated with PCB or PBB (5 and 10 ppm), the exception being T3 in the 5 ppm PCB treatment group. Had the PCB and PBB treatment-induced suppression of T4 and T3 been on the hypothalamo-pituitary axis, the response of the treated rats to exogenous TSH might have exceeded that of controls; however, the opposite occurred. Disappearance of injected doses of L [125I]T4 diminished as treatment concentrations of PCB or PBB increased. Disappearance slopes (r = 0.98) and fractional turnover rate constants (k) were decreased (t1/2 was lengthened) at each treatment level compared to the control values. The T4 distribution space (per 100 g BW) was expanded with increasing dosage by as much as 8-fold in the 50 pmm PCB treatment group. T4 MCRs were not increased by PCB or PBB treatment; thus, decreases in serum T3 and T4 were not caused by increased catabolism. T4 production rates were decreased at all treatment levels, but maximally 6-fold by 50 ppm PCB treatment. Together these data indicate that PCB-PBB-induced decreases in serum T3 and T4 result primarily from direct damage to the thyroid rather than any enhanced hepatic or other peripheral catabolism per se. Expanded T4 distribution space demonstrated that nonthyroid damage was also an important factor in reducing serum T4. Cell membrane damage associated with PCB-PBB intoxication may have expanded pools for T4 dilution. The findings are consistent with reported histological and ultrastructural damage caused by PCB and PBB. It also appears that TSH plays little role in PCB-PBB-induced hypothyroidism. PMID- 3036478 TI - Bovine granulosa cells produce basic fibroblast growth factor. AB - Cultured bovine granulosa cells express the gene encoding basic fibroblast growth factor (bFGF). The bFGF gene is transcribed into 7.0- and 3.7-kilobase mRNA transcripts which are apparently translated into 16,000 mol wt bFGF-like growth factor. The granulosa cell-derived bFGF is bioactive, i.e. it can stimulate the proliferation of capillary endothelial or granulosa cells. This mitogenic effect is prevented by specific neutralizing anti-bFGF antibodies. Our results indicate that bFGF derived from granulosa cells can act as both autocrine and paracrine growth factor, and they further suggest that the factor may be involved in the development of the rich vasculature of the theca interna of the follicle. PMID- 3036479 TI - A nuclear binding assay to assess the biological activity of steroid receptors in isolated animal and human tissues. AB - This paper describes a nuclear binding assay (NB assay) which measures not only the presence of a steroid receptor in a tissue, but also the quantity of that receptor which is biologically active or functional, i.e. able to bind to nuclear acceptor sites. The assay involves the isolation viable cells from tissues and their incubation with an excess of radiolabeled steroid to encourage the activation and nuclear binding of all cellular receptors. The nuclei are isolated under conditions that remove unactivated (unbound) steroid-receptor complexes. This NB assay demonstrates, in both animal and human steroid target tissues, a saturable, tissue- and steroid-specific, and temperature- and time-dependent nuclear binding of radiolabeled steroids. These properties support a receptor dependent nuclear binding of steroids. This assay is reproducible and requires relatively small amounts of tissue. The patterns of nuclear binding of the progesterone receptor, achieved with the assay in the avian oviduct model system, are shown to correlate with the nuclear binding of progesterone in vivo, the ability of the steroid to alter transcription, and the expression of a specific gene product, the protein avidin. The assay has been used to identify the existence of nonfunctional steroid receptors in endometrial and breast carcinomas. Therefore, this NB assay combined with the standard charcoal/hydroxylapatite methods of quantitating total cellular receptors should provide a means of assessing changes in the regulation of the biological activity of steroid receptors. Further, the assay should be useful to assess the ability of steroid analogs to properly activate their respective receptors for subsequent nuclear binding. PMID- 3036480 TI - Direct inhibitory effect of long term estradiol treatment on dopamine synthesis in tuberoinfundibular dopaminergic neurons: in vitro studies using hypothalamic slices. AB - The mechanism of the inhibitory effect of long term treatment with estradiol on dopamine synthesis in tuberoinfundibular dopaminergic (TIDA) neurons was studied by using hypothalamic slices from ovariectomized rats. Treatment with 2 mg estradiol valerate (EV) at a 3-week interval increased the weight of the anterior pituitary gland and the concentration of serum PRL. In vivo and in vitro dopamine synthesis in TIDA neurons were estimated in EV-treated animals by 3,4 dihydroxyphenylalanine (DOPA) accumulation in the median eminence after injections of 3-hydroxybenzylhydrazine (NSD 1015), a DOPA decarboxylase inhibitor, and after incubation of hypothalamic slices with NSD 1015, respectively. In vivo DOPA accumulation in the median eminence was less in EV treated rats than in control rats. The basal rate of in vitro DOPA accumulation in the median eminence of hypothalamic slices from EV-treated rats was lower than that in control rats. Ca2+-dependent DOPA accumulation in the median eminence, determined by incubation in medium containing depolarization agents such as 50 mM K+ and veratridine, was decreased in EV-treated rats. Furthermore, cAMP-dependent DOPA accumulation, determined by incubation with Bu2cAMP or forskolin, was also suppressed in EV-treated rats. The decreased depolarization-induced DOPA accumulation in the median eminence recovered after cessation of EV treatment. Hyperprolactinemia lasting for 6 weeks, achieved by transplantation of anterior pituitaries under the kidney capsule, increased the rate of depolarization induced DOPA accumulation in the median eminence. On the other hand, EV treatment was effective in inhibiting depolarization-induced DOPA accumulation in hypophysectomized rats regardless of the presence of anterior pituitary transplants. These results suggest that chronically administered estradiol inhibits dopamine synthesis in TIDA neurons via a direct action on the hypothalamus and overcomes the facilitatory action of PRL on dopamine synthesis; and estradiol inhibits all three distinct systems that regulate basal, Ca2+ dependent, and cAMP-dependent dopamine synthesis in TIDA neurons. PMID- 3036481 TI - Interaction of insulin-like growth factor I with porcine thyroid cells cultured in monolayer. AB - The interaction of insulin-like growth factor I (IGF-I) with porcine thyroid cells cultured in monolayer was studied. Specific binding of [125I]iodo-IGF-I to thyroid cells was a reversible process dependent on the time and temperature of incubation. A steady state was achieved in 18 h at 4 C and averaged 14.2 +/- 2% (mean +/- SD)/10(6) cells. Binding of [125I]iodo-IGF-I was inhibited by unlabeled IGF-I; half-maximal inhibition occurred at concentrations of 2-5 ng/ml. Multiplication-stimulating activity (rat IGF-II) and pork insulin had relative potencies of 1:20 and 1:300 compared with IGF-I. Scatchard analysis of binding data revealed a single class of IGF-I receptors with a Ka of 4.3 X 10(10) M-1, 49,000 binding sites were estimated per cell. Affinity cross-linking and autoradiography demonstrated the presence of type I IGF receptors. Thyroid cells also had specific receptors for insulin, but specific binding of [125I]iodoinsulin (2.03 +/- 0.03%/10(6) cells) was much lower than that of [125I]iodo-IGF-I. Preincubation of thyroid cells with IGF-I or insulin caused a concentration-dependent decrease in [125I]iodo-IGF-I binding due to an apparent loss of receptors. Preincubation with epidermal growth factor, fibroblast growth factor, platelet-derived growth factor, or TSH did not alter subsequent binding of [125I]iodo-IGF-I. Low concentrations of IGF-I stimulated DNA synthesis and proliferation of thyroid cells and acted synergistically with epidermal growth factor. Multiplication-stimulating activity and insulin had relative potencies in stimulating DNA synthesis comparable to their abilities to inhibit the binding of [125I]iodo-IGF-I to thyroid cells, suggesting that their effects are mediated primarily by IGF-I receptors. Preincubation with IGF-I did not alter cAMP responsiveness to TSH. We, thus, demonstrated the presence of functional and regulated IGF-I receptors on porcine thyroid cells. PMID- 3036482 TI - A growth stimulatory effect of iodide is suggested by its effects on c-myc messenger ribonucleic acid levels, [3H]thymidine incorporation, and mitotic activity of porcine follicle cells in suspension culture. AB - In the present study the effect of iodide on thyroid cell growth was investigated in primary suspension cultures of porcine thyroid cells capable of organifying iodide. The addition of a high dose of iodide (10(-4) M) to such cultures caused a marked increase in c-myc mRNA levels, [3H]thymidine incorporation, and mitotic activity. The incorporation of [3H]thymidine started 30-36 h after the addition of iodide. The stimulatory effect was abolished by a simultaneous incubation with methimazole. The concentration dependence of the iodide-induced stimulation of [3H]thymidine incorporation was similar to that of an inhibitory effect on adenylate cyclase activity. W-7, an inhibitor of calmodulin activity, as well as epinephrine, agents that reduce cAMP levels, also stimulated [3H]thymidine incorporation. Moreover, the stimulatory effect of iodide was reduced in the presence of forskolin. The results suggest that an organic form of iodine stimulates thyroid cell growth by reducing cAMP levels and demonstrate the presence of a growth stimulatory pathway in porcine thyroid cells that is independent of exogenous polypeptide growth factors or hormones. PMID- 3036483 TI - Alpha 2-adrenergic potentiation of adenosine-stimulating effect on glucagon secretion. AB - Previous studies from our laboratory showed 1) that adenosine (1.65 microM), a substance released by tissues in energy-deficient states, stimulated glucagon secretion by activation of A2 purinergic receptors, and 2) that this effect was potentiated by a low substimulating concentration of epinephrine through activation of alpha-adrenergic receptors. The present work was undertaken to assess the subtype of alpha-adrenergic receptor involved in this potentiation. Therefore, we used adrenergic blockers and agonist drugs more specific for alpha 1- or alpha 2-adrenergic receptors. The potentiating effect of epinephrine (0.01 microM) on glucagon secretion induced by adenosine (1.65 microM) was not prevented by an alpha 1-adrenergic blocker, prazosine (6 microM), but was suppressed by an alpha 2-adrenergic blocker, yohimbine (0.6 microM). The implication of alpha 2-adrenergic receptors in the potentiating effect was confirmed by the use of selective alpha 1- or alpha 2-adrenergic agonist drugs. Indeed, clonidine (0.01 microM), an alpha 2-agonist, ineffective per se, potentiated, whereas phenylephrine (0.01 microM), an alpha 1-agonist, had no effect on glucagon secretion induced by adenosine. We conclude that the potentiation by epinephrine of adenosine-induced glucagon secretion is mediated by alpha 2-adrenergic receptor activation. A potentiation between the effects of A2 purinergic and alpha 2-adrenergic agonists may be of physiological relevance in stressful energy-deficient states, when an increase in glucagon secretion is necessary. PMID- 3036484 TI - Intracellular Ca2+-dependent protein kinase C activation mimics delayed effects of thyrotropin-releasing hormone on clonal pituitary cell excitability. AB - Biochemical and spectrophotometric studies of second messenger pathways transducing TRH signals in clonal pituitary (GH) cells have shown that TRH induces rapid turnover of phosphoinositides and changes in cytoplasmic Ca2+ as well as activation of protein kinase C (PKC) and secretion of PRL. Here we have used classical microelectrode and contemporary patch pipette recording techniques under current-clamp conditions to compare the effects of TRH receptor-coupled stimulation with direct activation of PKC on the excitability of GH3/B6 cells. With high resistance microelectrodes TRH induced a complex sequence of changes in membrane properties consisting of an initial 20- to 30-mV hyperpolarization associated with an increase in membrane conductance lasting less than a minute, followed by several minutes of low amplitude fluctuations and action potential activity superimposed on a modest increase in input resistance. Active phorbol ester induced a slowly developing hyperpolarization of about 5 mV and a modest increase in input resistance, followed by several minutes of low amplitude fluctuations and spontaneous action potential activity. Both the peptide- and phorbol ester-evoked changes in excitability were attenuated or completely lost during patch recordings in the whole cell mode. Dilute aqueous lysates of the clone restored various phases of the electrical response. The low amplitude fluctuations and action potential activity phase could be induced by either TRH or phorbol ester if the cells were dialyzed with intracellular electrolyte containing PKC and at least 50 nM Ca2+. These results demonstrate that the phosphoinositide/PKC circuit activated by TRH in clonal pituitary cells has electrically detectable effects on cell excitability, and these help to explain TRH's actions on electrical activity. PMID- 3036485 TI - Analysis of ligand-binding to the kringle 4 fragment from human plasminogen. AB - The interaction of the isolated human plasminogen kringle 4 with the four omega amino acid ligands epsilon-aminocaproic acid (epsilon ACA), N alpha-acetyl-L lysine (AcLys), trans-aminomethyl(cyclohexane)carboxylic acid (AMCHA) and p benzylaminesulfonic acid (BASA) has been further characterized by 1H-NMR spectroscopy at 300 and 600 MHz. Pronounced high-field shifts, reaching approximately 3 ppm, are observed for AMCHA resonances upon binding to kringle 4, which underscores the relevance of ligand lipophilic interactions with aromatic side chains at the binding site. Ligand titration curves for the nine His and Trp singlets found in the kringle 4 aromatic spectrum reveal a striking uniformity in the kringle response to the various ligands. The average binding curves exhibit a clear Langmuir absorption isotherm saturation profile and the data were analyzed under the assumption of one (high affinity) binding site per kringle. Equilibrium association constants (Ka) and first order dissociation rate constants (k off) were derived from linearized expressions of the Langmuir isotherm and of the spectral line-shapes, respectively. The results for the four ligands, at approximately 295 K, pH 7.2, indicate that: AMCHA exhibits the strongest binding (Ka = 159 mM-1) and epsilon ACA the weakest (Ka = 21 mM-1) with AcLys and BASA falling in between; epsilon ACA dissociates readily (k off = 5.3 X 10(3) s-1) and AMCHA associates the fastest (k on = 2.0 X 10(8) M-1 s-1) while the kinetics for BASA exchange is relatively slow (k off = 0.8 X 10(3) s-1, k on = 0.6 X 10(8) M-1 s-1); the ligand-binding kinetics is close to diffusion-controlled. PMID- 3036486 TI - The structure of ubiquitinated histone H2B. AB - Ubiquitinated histone H2B (uH2B) has been purified from both calf and pig thymus by exclusion chromatography in 7 M urea. Digestion of uH2B with Staphylococcus aureus V8 protease yielded the peptide 114-125 containing the ubiquitin moiety. Further digestion of this peptide with trypsin removed the ubiquitin and three H2B residues from the N-terminus. Edman degradations of both peptides established that ubiquitin is attached to the epsilon-amino group of lysine 120 in both calf and pig uH2B by an iso-peptide bond to the C-terminal glycine 76 of ubiquitin. PMID- 3036487 TI - Discrete elements within the SV40 enhancer region display different cell-specific enhancer activities. AB - The SV40 enhancer contains three genetically defined elements, called A, B and C, that can functionally compensate for one another. By using short, synthetic DNA oligonucleotides, we show that each of these elements can act autonomously as an enhancer when present as multiple tandem copies. Analysis of a progressive series of B element oligomers shows a single element is ineffective as an enhancer and that the activity of two or more elements increases with copy number. Assay in five different cell lines of two separate enhancers containing six tandem copies of either the B or C element shows that these elements possess different cell specific activities. Parallel oligomer enhancer constructs containing closely spaced double point mutations display no enhancer activity in any of the cell lines tested, indicating that these elements represent single units of enhancer function. These elements contain either a 'core' or 'octamer' consensus sequence but these consensus sequences alone are not sufficient for enhancer activity. The different cell-specific activities of the B and C elements are consistent with functional interactions with different trans-acting factors. We discuss how tandem duplication of such dissimilar elements, as in the wild-type SV40 72-bp repeats, can serve to expand the conditions under which an enhancer can function. PMID- 3036488 TI - Differential promoter utilization by the bovine papillomavirus in transformed cells and productively infected wart tissues. AB - Expression of the 'late' genes of bovine papillomavirus type 1 (BPV-1) occurs only in the differentiated keratinocytes of the productively infected fibropapilloma. A detailed analysis of viral transcription in the fibropapilloma was performed and compared to BPV-1 specific transcription in transformed C127 cells. A cDNA library was constructed from bovine fibropapilloma mRNA using the method of Okayama and Berg. Analysis of full length cDNAs showed that the majority of viral transcripts in the fibropapilloma have 5' termini near nt 7250 and utilize a common splice donor site at nt 7385. This mRNA start site was confirmed by the combination of primer extension and nuclease S1 analyses; it is not utilized in the BPV-1-transformed C127 cell, thus identifying it as a wart specific, 'late' promoter. Upstream of this mRNA start site is a tandemly repeated sequence element homologous to the SV40 late promoter sequence GGTACCTAACC, which has been shown to be important for the efficient utilization of the SV40 major late start site. Two additional mRNA start sites at nt 7185 and nt 7940 in the long control region (LCR) were identified and were found to be used in bovine warts as well as in BPV-1-transformed mouse cells. The promoter region upstream of the nt 7940 mRNA start site contains the E2 responsive enhancer mapping between nt 7611 and nt 7805 [Spalholz, B.A., Lambert, P.F., Yee, C. and Howley, P.M. (1987) J. Virol., in press].(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3036490 TI - Both ATP and an energized inner membrane are required to import a purified precursor protein into mitochondria. AB - We have investigated the energy requirement of mitochondrial protein import with a simplified system containing only isolated yeast mitochondria, energy sources and a purified precursor protein. This precursor was a fusion protein composed of 22 residues of the cytochrome oxidase subunit IV pre-sequence fused to mouse dihydrofolate reductase. Import of this protein required not only an energized inner membrane, but also ATP. ATP could be replaced by GTP, but not by CTP, TTP or non-hydrolyzable ATP analogs. Added ATP did not increase the membrane potential of respiring mitochondria; it supported import even if the proton translocating mitochondrial ATPase and the entry of ATP into the matrix were blocked. We conclude that ATP exerts its effect on mitochondrial protein import outside the inner membrane. PMID- 3036489 TI - Amplification of the X-linked Drosophila chorion gene cluster requires a region upstream from the s38 chorion gene. AB - Genomic sequences controlling follicle cell-specific amplification of the X linked Drosophila chorion gene cluster were mapped by P element-mediated transformation. Several DNA fragments containing the s38 gene and flanking sequences induced tissue-specific amplification, although replication levels were subject to position effects. Deletion analysis identified a 467-bp region upstream from the s38 transcription start site that contained sequences essential in cis for amplification. The essential region shared 32 bp of imperfect sequence homology with a previously identified region necessary for third chromosome chorion gene cluster amplification. This homologous segment contained a repetitive motif consisting of perfect and imperfect AATAC repeats; it was localized near the boundary of the essential domain since most, but not all, the repeats could be deleted without eliminating transposon-induced amplification. The repetitive region was not required for developmentally regulated s38 transcription, therefore our results identified at least one element required for amplification but not for chorion gene transcription. The homologous repetitive sequences within the amplification-essential regions may constitute part of the replication origins used to differentially replicate the two chorion domains during oogenesis. PMID- 3036491 TI - Efficient splicing of two yeast mitochondrial introns controlled by a nuclear encoded maturase. AB - bI4 maturase encoded by the fourth intron of the yeast mitochondrial cytochrome b gene, controls the splicing of both the fourth intron of the cytochrome b gene and the fourth intron of the gene encoding subunit I of cytochrome oxidase. It has been shown previously that a cytoplasmically translated hybrid protein composed of the pre-sequence of subunit 9 of Neurospora ATPase fused to a part of the bI4 maturase can be guided to mitochondria where it could compensate maturase deficiencies. This in vivo complementation of maturase mutants can be easily estimated by restoration of respiration. This work examines the efficiency of different bI4 maturase constructions to restore respiration in different yeast maturase-deficient strains. It is shown that the N-terminal end of the bI4 maturase plays a crucial role in the maturase activity. Moreover, the 12 N terminal amino acids of the mitochondrial outer membrane protein constitute the most efficient mitochondrial targeting sequence in this system. Surprisingly enough, it was found that the cytoplasmically translated bI4 maturase containing the 254 C-terminal amino acid coded by the intron open reading frame can complement maturase mutations without any added mitochondrial-targeting sequence. PMID- 3036492 TI - Transcriptional termination at a fully rho-independent site in Escherichia coli is prevented by uninterrupted translation of the nascent RNA. AB - We have examined the possibility that translation reading through a fully rho independent transcriptional terminator in Escherichia coli might prevent termination, as already established for rho-dependent terminators. Plasmids were constructed with and without interposition of the rho-independent coliphage T7 'early' terminator between a promoter and galK. Our constructions ensured either that there was no upstream translation, or that translation (initiated at the galE ribosome binding site) stopped upstream of, or at the normal position (the T7 gene 1.3 stop codon) with respect to, the transcriptional terminator; or else downstream of both this stop codon and the terminator. Our galactokinase enzyme and mRNA measurements on strains harbouring these plasmids indicate that 'readthrough translation' eliminates transcriptional termination at the T7 site. This effect is suppressed if the rate of ribosome movement is reduced with fusidic acid. PMID- 3036493 TI - Calcitonin gene-related peptide elevates cyclic AMP levels in chick skeletal muscle: possible neurotrophic role for a coexisting neuronal messenger. AB - Recent immunocytochemical studies have shown that calcitonin gene-related peptide (CGRP) coexists with the neurotransmitter acetylcholine in spinal motoneurons of the chick. Moreover, CGRP causes an increase in the number of acetylcholine receptors on the surface of cultured chick myotubes. CGRP might thus serve as one of the signals by which motoneurons regulate endplate development. In a search for the second messengers involved, we now demonstrate that CGRP stimulates accumulation of cyclic AMP (cAMP) in cultured chick myotubes. This effect is, at least in part, mediated by an increase in cAMP synthesis, as the peptide also activates adenylate cyclase in chick muscle membranes. Nanomolar concentrations of CGRP elicit significant increases in both cellular cAMP levels and acetylcholine receptor numbers. The present findings suggest that cAMP is one of the second messengers which mediate the increase in acetylcholine receptor number elicited by CGRP. Furthermore, CGRP might be implicated in other trophic actions mediated by cAMP in skeletal muscle cells. PMID- 3036494 TI - A temperature-sensitive mutant of Abelson murine leukemia virus confers inducibility of IgM expression to transformed lymphoid cells. AB - Lymphoid cell lines were isolated that were inducible for the expression of surface immunoglobulin by shift from 35.5 to 39.5 degrees C after infection of mouse bone marrow cells with a mutagen-treated Abelson murine leukemia virus. Virus produced by one of the cell lines (ts49) transmitted the temperature sensitive phenotype to new lymphoid transformants as well as to NIH/3T3 cells. In addition, the tyrosine autophosphorylating activity of the p120gag-abl protein synthesized in ts49-transformed cells was found to be temperature-sensitive. Shift experiments using ts49-transformed lymphoid cells showed that at 39.5 degrees C they synthesize increased amounts of mu and kappa chain RNA and protein, and that they can be further induced to secrete IgM when treated with lipopolysaccharide. PMID- 3036495 TI - Identification of the HPV-16 E6 protein from transformed mouse cells and human cervical carcinoma cell lines. AB - Human cervical carcinoma cell lines that harbor human papillomavirus (HPV) have been reported to retain selectively and express HPV sequences which could encode viral E6 and E7 proteins. The potential importance of HPV E6 to tumors is suggested further by the observation that bovine papillomavirus (BPV) E6 can induce morphologic transformation of mouse cells in vitro. To identify HPV E6 protein, a polypeptide encoded by HPV-16 E6 was produced in a bacterial expression vector and used to raise antisera. The antisera specifically immunoprecipitated the predicted 18-kd protein in two human carcinoma cell lines known to express HPV-16 RNA and in mouse cells morphologically transformed by HPV 16 DNA. The 18-kd E6 protein was distinct from a previously identified HPV-16 E7 protein. The HPV-16 E6 antibodies were found to be type specific in that they did not recognize E6 protein in cells containing HPV-18 sequences and reacted weakly, if at all, to BPV E6 protein. The results demonstrate that human tumors containing HPV-16 DNA can express an E6 protein product. They are consistent with the hypothesis that E6 may contribute to the transformed phenotype in human cervical cancers that express this protein. PMID- 3036496 TI - The control of hypertension with dibutyryl cyclic AMP. AB - The effects of the rapid infusion of large doses of dibutyryl cyclic AMP (DBcAMP) were studied to clarify the clinical usefulness of its vasodilating action in 32 middle-aged patients, who underwent various types of surgery and developed systolic hypertension of over 160 mmHg during general anaesthesia. DBcAMP was given i.v. with an infusion pump at a rate of 0.6 mg kg-1 min-1 for 20 min. In all patients just after the infusion, systolic arterial pressure decreased from 174.0 +/- 20.7 to 129.0 +/- 23.9 mmHg, diastolic pressure decreased from 93.1 +/- 13.4 to 64.8 +/- 13.3 mmHg, heart rate increased from 81.2 +/- 15.7 to 91.5 +/- 19.5 beats min-1, and urine volume increased from 69.4 +/- 54.8 to 182.7 +/- 143.5 ml h-1. In three patients, cardiac index increased from 3.44 to 4.24 l min 1 m-2. In seven patients, tachycardia exceeding 120 beats min-1 developed. DBcAMP was also effective in patients with a history of hypertension. The strongest antihypertensive effect was observed in patients anaesthetized with nitrous oxide oxygen and enflurane. We speculate that DBcAMP is useful to control hypertension and may be particularly indicated in patients with cardiac failure, renal disorders and essential hypertension. PMID- 3036498 TI - In vitro activity of (S)-9-(3-hydroxy-2-phosphonylmethoxypropyl)adenine against newly isolated clinical varicella-zoster virus strains. AB - The highly potent and selective anti-DNA virus agent (S)-9-(3-hydroxy-2 phosphonylmethoxypropyl)adenine [(S)-HPMPA] was found to inhibit in vitro the replication of a number of clinical varicella-zoster virus strains within the concentration range of 0.63-5.7 ng/ml. With a mean 50% inhibitory concentration of 1.8 ng/ml and selectivity index of 29000, (S)-HPMPA is one of the most potent and most selective varicella-zoster virus inhibitors discovered to date. PMID- 3036499 TI - Comparative in vitro activity of the new cyclic lipopeptide LY146032 against Corynebacterium species. AB - Agar dilution and time-kill techniques were used to assess the in vitro activity of LY146032 and several other antibacterial agents against Corynebacterium spp. The activity of LY146032 was similar to that of vancomycin and teicoplanin. Synergistic killing by a combination of LY146032 and gentamicin could be demonstrated under certain carefully controlled conditions. PMID- 3036497 TI - Development of a monoclonal antibody specific for serotype 3 rotavirus strains. AB - Monoclonal antibodies were prepared against serotype 3 simian rotavirus SA11. Antigenic analysis of 18 hybridoma cell lines secreting monoclonal antibodies by radioimmunoprecipitation and Western blot revealed that seven monoclonals were directed against the major inner capsid protein VP6, four against VP3, an outer capsid protein with hemagglutinating activity, and one against VP7, the main outer capsid protein of the virus. The specificity of six monoclonals could not be determined. One monoclonal (1P14E2) directed against VP3 showed serotype 3 specific neutralizing activity. This monoclonal, which recognized only serotype 3 viruses in an enzyme-linked immunosorbent assay, could be useful in assays for serotyping rotavirus directly in stool samples. PMID- 3036500 TI - Effect of abscess milieu on bactericidal activity of LY146032 against staphylococci. AB - The antistaphylococcal activity of LY146032 was tested in human pus in vitro and in a murine abscess model in vivo. The drug was not degraded by pus containing beta-lactamase and had equally good or better activity than nafcillin or vancomycin against Staphylococcus aureus or Staphylococcus epidermidis in vitro and in vivo. Its activity was slightly enhanced when used in combination with rifampin and tobramycin. PMID- 3036501 TI - Susceptibility of Clostridium difficile to LY146032. PMID- 3036502 TI - Comparison of bronchial washing, brushing and biopsy for diagnosis of pulmonary tuberculosis. AB - The diagnostic yields of bronchial washings, bronchial brushings and lung biopsy specimens were compared in 50 patients with positive Mycobacterium tuberculosis cultures. The number of positive results obtained with cultures of bronchial brushings was significantly higher than that with bronchial washings (p less than 0.001). The histological study of biopsy lung material improved the rate of immediate or rapid diagnosis of tuberculosis (p less than 0.001). PMID- 3036503 TI - Antibody response to individual cytomegalovirus structural proteins in different groups of subjects. AB - The antibody response to cytomegalovirus structural polypeptides in sera from three groups of acutely infected subjects was analyzed. Differences in number and types of polypeptides were noted. Immunoblotting could be used to distinguish sera from patients with acute cytomegalovirus hepatitis from convalescent sera by the detection of antibody to viral proteins of 82, 66, 62, and 55 kilodalton molecular weight at a high serum dilution. PMID- 3036504 TI - [D-Phe4]peptide histidine-isoleucinamide ([D-Phe4]PHI), a highly selective vasoactive-intestinal-peptide (VIP) agonist, discriminates VIP-preferring from secretin-preferring receptors in rat pancreatic membranes. AB - The capacity of vasoactive intestinal peptide (VIP), peptide histidine isoleucinamide (PHI), secretin, and a series of analogs to discriminate between VIP-preferring and secretin-preferring receptors that coexist in rat pancreatic plasma membranes was evaluated by their ability to inhibit [125I]iodo-VIP and [125I]iodo-secretin binding and to activate adenylate cyclase. VIP, the VIP analogs [D-His1]VIP, [D-Ser2]VIP, [D-Asp3]VIP and [D-Ala4]VIP, PHI, [D-Phe4]PHI, and secretin inhibited the binding of both ligands in a concentration range of 10(-11) M to 10(-5) M and with a selectivity factor varying from 18,000 to 0.1. The only exception was [D-Phe4]PHI that inhibited 125I-VIP binding only, with an IC50 of 7 nM, and with no inhibition of 125I-secretin binding at 10 microM. The peptides tested stimulated adenylate cyclase in the same membranes and the slope of the dose-effect curves indicated that all peptides, except [D-Phe4]PHI, interacted with at least two classes of receptors: VIP-preferring and secretin preferring receptors. By contrast, the dose-effect curve of [D-Phe4]PHI activation of adenylate cyclase was monophasic and competitively modified by [D Phe2]VIP (a VIP antagonist) but not by secretin(7-27) (a secretin antagonist), indicating an interaction with VIP-preferring receptors only. Thus, [D-Phe4]PHI appears to be a highly selective tool to characterize these receptors. PMID- 3036505 TI - Signal sequence and DNA-mediated expression of human lysosomal alpha galactosidase A. AB - Twelve complementary DNA clones for human lysosomal alpha-galactosidase A were isolated from an Okayama-Berg library constructed from SV40-transformed human fibroblasts. The identity of these clones was confirmed by complete colinearity of the nucleotide-deduced amino acid sequence with that determined by direct chemical sequencing of human placental alpha-galactosidase A. Hybridization of the alpha-galactosidase A cDNA to genomic DNA from individuals with varying numbers of X chromosomes as well as from interspecies somatic-cell hybrids showed only a single locus in the genome at Xq 13.1-Xq 22. One cDNA clone (pcD-AG210) contained the complete coding sequence for both the signal peptide and mature alpha-galactosidase A. The signal peptide of 31 amino acids contains the expected hydrophobic domains consisting of Leu-Gly-Cys-Ala-Leu-Ala-Leu and Phe-Leu-Ala-Leu Val and has Ala at the signal peptidase cleavage site. Twelve out of fifteen G residues flanking the 5' end of the cDNA in pcD-AG210 were removed and the truncated fragment was ligated into the original vector. This construct, pcD AG502, encoded enzymatically active human alpha-galactosidase A in monkey COS cells. PMID- 3036506 TI - Molecular characterization of transposable-element-associated mutations that lead to constitutive L-ornithine aminotransferase expression in Saccharomyces cerevisiae. AB - The cargB or CAR2 gene, coding for ornithine aminotransferase, was isolated by functional complementation of a cargB- mutation in Saccharomyces cerevisiae. It was used as a hybridization probe to analyse RNA and chromosomal DNA from four strains bearing cis-dominant regulatory mutations leading to constitutive, mating type-dependent, ornithine aminotransferase synthesis. The four mutations appear to be insertions. Their size and restriction pattern suggested that they were transposable elements, Ty1. All were inserted in the same orientation with respect to the cargB gene. We cloned the cargB gene with its associated insertion from two constitutive mutants (cargB+ Oh-1 and cargB+ Oh-2). We determined the sequence of the cargB 5' region from the wild-type gene and from the two mutated genes. The DNA sequences of the extremities of the two insertions were very homologous but not identical and were similar to previously reported Ty1 element direct repeats (delta). The same five-base-pair sequence, ATATA, was found at both ends of both Ty1 elements, indicating that both Ty1 were transposed to the same site. This site is located 115 base pairs upstream from the putative cargB coding region. The 5' end of cargB transcript as determined by S1 mapping was the same in the wild-type strain and in the four mutants. The cargB transcript was not detected in the wild-type strain grown under non-induced conditions, while under the same conditions it was present in all four mutants. PMID- 3036507 TI - Nucleotide sequence of the gene encoding the 11-kDa subunit of the ubiquinol cytochrome-c oxidoreductase in Saccharomyces cerevisiae. AB - The nucleotide sequence of the gene encoding the 11-kDa subunit VIII of the ubiquinol-cytochrome-c oxidoreductase in Saccharomyces cerevisiae has been determined. The coding sequence has a length of 330 bp and is preceded at a distance of 361 bp by another reading frame, coding for a protein of as yet unknown function. The 11-kDa gene is transcribed independently of the URFx gene and transcription of both is sensitive to catabolite repression. Multiple 5' and 3' termini of transcripts of the gene for the 11-kDa subunit were identified by S1 nuclease protection analysis of DNA X RNA hybrids. The 5' termini map 52 +/- 2 and 60 +/- 2 nucleotides upstream of the initiation codon whereas the 3' termini map 336 +/- 2 and 350 +/- 2 nucleotides downstream of the stop codon. The subunit VIII reading frame encodes a protein with a molecular mass of 12.4 kDa and a polarity of 37.6%. It is predicted to contain a high content of beta-sheet segments, which may be capable of forming a barrel-like structure in a lipid bilayer. A comparison of the sequence with those of the small subunits of the beef heart complex reveals similarity with the 9.5-kDa subunit VII (core-linked protein) characterized by Borchart et al. (1986) FEBS Lett. 200, 81-86. The significance of this is discussed. PMID- 3036508 TI - Fructofuranose 2-phosphate is the product of dephosphorylation of fructose 2,6 bisphosphate. AB - Using comparative ion-exchange chromatography on Dowex 1X4, the product of dephosphorylation of fructose 2,6-bisphosphate with purified yeast fructose-2,6 bisphosphate 6-phosphohydrolase, was shown to be identical to the furanose form of fructose 2-phosphate prepared by chemical synthesis according to Pontis and Fischer [Biochem. J. 89, 452-459 (1963)]. As expected for the furanose form of fructose 2-phosphate, the enzymatically formed product consumes 1 mol periodate/mol fructose 2-phosphate, whereas the chemically synthesized pyranose form consumes 2 mol periodate/mol. In addition, it is shown that the enzymatic product behaves identically to the furanose, not the pyranose, form of fructose 2 phosphate in hydrolysis of the ester bond at pH 4 and 37 degrees C, as described previously for the chemically synthesized compounds [Pontis and Fischer (1963) vide supra]. PMID- 3036509 TI - Covalent flavinylation of 6-hydroxy-D-nicotine oxidase analyzed by partial deletions of the gene. AB - The expression of the enzymatically active 6-hydroxy-D-nicotine oxidase (6-HDNO) from Arthrobacter oxidans requires the covalent attachment of FAD to the polypeptide chain. How this modification takes place and at what time during the synthesis of the polypeptide is not known. We investigated the possibility of cotranslational flavinylation by generating various deletions of the 6-HDNO gene carried on appropriate plasmid vectors. The polypeptides expressed from these plasmids were analyzed for their ability to incorporate [14C]FAD covalently in an Escherichia coli-derived coupled transcription/translation system. The data show that removal of approximately 40% from the carboxy-terminal part of the 6-HDNO polypeptide did not inhibit the covalent flavinylation of the truncated protein. A fusion protein, consisting of the truncated 6-HDNO polypeptide and the beta lactamase of pBR322, was also covalently flavinylated. The amino acid sequence surrounding the histidine residue, assumed to bind FAD, was shown to be situated approximately 70 amino acid residues from the amino-terminal end of the 6-HDNO polypeptide. Removal of the first 30 amino acids did not abolish covalent flavinylation. Flavinylation could no longer be detected, however, if a short amino acid sequence, consisting of seven residues, replaced the amino acid sequence upstream of the histidine. These findings prove, in our opinion, that cotranslational flavinylation takes place in the synthesis of 6-HDNO. PMID- 3036510 TI - Isolation and some properties of the site-specific endonuclease and methylase Bme2161 from Bacillus megaterium 216. AB - The site-specific endonuclease Bme2161 was isolated as a homogeneous preparation by chromatography on phosphocellulose, hydroxyapatite and heparin-agarose. The molecular mass of the enzyme, determined by gel filtration and by electrophoresis under denaturing conditions, was found to be 60 kDa and 30 kDa respectively. These data indicate that the native enzyme consists of two identical subunits. The enzyme recognized the decreases pentanucleotide sequence 5'-GGACC-3' X 3' CCTGG-5' and cleaves the sequence as indicated by arrows. The increases optimal concentration for endonuclease reaction is 6-7 mM Mg2+. The endonuclease relaxes its specificity in the presence of glycerol or dimethyl sulfoxide at low Mg2+ concentration (1-3 mM). Methylase Bme2161, which protects DNA against endonuclease Bme2161 action by DNA methylation, was isolated from the same bacterial strain. PMID- 3036511 TI - Ouabain-binding site of (Na+ + K+)-ATPase in right-side-out vesicles has not an externally accessible SH group. AB - The fluorescing sulfhydryl reagent N-(7-dimethylamino-4 methylcoumarinyl)maleimide (DACM) inactivates purified (Na+ + K+)-ATPase at 20 microM. This inactivation results in a decrease of the ouabain-binding capacity of the enzyme. Treatment of (Na+ + K+)-ATPase, embedded in right-side-out oriented vesicles, by DACM does not affect ouabain binding to the enzyme. Incorporation of DACM into the alpha subunit of (Na+ + K+)-ATPase embedded in right-side-out vesicles is also not affected by the presence or absence of 100 microM ouabain. It is therefore concluded that a sulfhydryl group does not reside within the ouabain-binding site of (Na+ + K+)-ATPase. PMID- 3036512 TI - Isolation and characterization of ubiquinol oxidase complexes from Paracoccus denitrificans cells cultured under various limiting growth conditions in the chemostat. AB - To obtain more information about the composition of the respiratory chain under different growth conditions and about the regulation of electron-transfer to several oxidases and reductases, ubiquinol oxidase complexes were partially purified from membranes of Paracoccus denitrificans cells grown in carbon-source limited aerobic, nitrate-limited anaerobic and oxygen-limited chemostat cultures. The isolated enzymes consisted of cytochromes bc1, c552 and aa3. In comparison with the aerobic ubiquinol oxidase complex, the oxygen- and nitrate-limited ones contained, respectively, less and far less of the cytochrome aa3 subunits and the anaerobic complex also contained lower amounts of cytochrome c552. In addition, extra haem-containing polypeptides were present with apparent Mr of 14,000, 30,000 and 45,000, the former one only in the anaerobic and the latter two in both the anaerobic and oxygen-limited preparations. This is the first report describing four different membrane-bound c-type cytochromes. The potentiometric and spectral characteristics of the redox components in membrane particles and isolated ubiquinol oxidase fractions were determined by combined potentiometric analysis and spectrum deconvolution. Membranes of nitrate- and oxygen-limited cells contained extra high-potential cytochrome b in comparison with the membranes of aerobically grown cells. No difference was detected between the three isolated ubiquinol oxidase complexes. Aberrances with already published values of redox potentials are discussed. PMID- 3036513 TI - Subfractionation and characterization of soluble c-type cytochromes from Paracoccus denitrificans cultured under various limiting conditions in the chemostat. AB - Soluble c-type cytochromes were partially purified from Paracoccus denitrificans cells grown in succinate- and methanol-limited aerobic, nitrate-limited anaerobic and oxygen-limited chemostat cultures. Five c types could be distinguished with the following apparent molecular masses, absorption maxima and midpoint potentials. (a) 9.2 kDa, 549 nm and +190 mV; (b) 14 kDa, 549 nm and +227 mV; (c) 22 kDa, 552 nm and +190 mV; (d) 30 kDa, 552.7 nm and +160 mV; (e) 45 kDa, a dihaem: 555 nm, +128 mV and 551 nm, -163 mV. The 14-kDa polypeptide was present under all growth conditions examined and most probably is the already well characterized cytochrome c550. In methanol-limited grown cells three additional cytochromes were found, the 9.2-kDa, 22-kDa and 30-kDa ones. Under oxygen-limited conditions the 45-kDa and under anaerobic growth conditions small quantities of the 30-kDa and 45-kDa cytochromes c were present. Based on the apparent molecular masses the 14-kDa, 22-kDa, 30-kDa and 45-kDa cytochromes may also be present in membrane-fractions. PMID- 3036514 TI - Internal acidification and cAMP increase are not correlated in Saccharomyces cerevisiae. AB - Addition of glucose to a yeast suspension can produce both an increase in the level of cAMP and a decrease in the intracellular pH. This observation led to the idea that internal acidification triggers the cAMP increase. We have tested this hypothesis using different approaches. To study the effect of sugar metabolism on internal pH we added to the yeast either glucose or a sugar, like xylose, that cannot be phosphorylated. We also utilized yeast strains lacking hexose kinases or phosphoglucose isomerase. We found that phosphorylation of the sugar added is a requisite for internal acidification but not for the cAMP increase. Internal acidification is due to an imbalance between the rate of the metabolic reactions that generate protons and the rate at which protons can be pumped out of the cell. We have manipulated the excretion of protons by using yeast harvested at different phases of growth and resuspended in a medium with or without added K+. Addition of glucose produced a marked drop in internal pH only when the yeast was harvested in the stationary phase of growth and transferred to a medium without added K+. In contrast an increase in cAMP was observed in all situations. We conclude that in yeast there is no correlation between internal acidification and cAMP increase. PMID- 3036515 TI - Cooperative interaction of multiple DNA elements in the human interferon-beta promoter. AB - Analysis of promoter mutants and hybrids in permanently transformed murine L cells reveals several regulatory DNA sequence elements in the 5' flanking region of the human interferon-beta gene, which together constitute the inducible promoter. The elements consist almost exclusively of purine runs in the region 111 to -1. Deletion of single elements reduces the expression capacity drastically, whereas duplication leads to a synergism of inducible expression. These elements act together in a cooperative way to achieve high inducibility. Natural and mutant promoter fragments containing these elements impose inducibility on a heterologous promoter. However, typical enhancer activity in this system is not observed. PMID- 3036516 TI - Purification and properties of the soluble cytochromes c-550 and c-556 from the bacterium Aquaspirillum itersonii. AB - Two c-type cytochromes were isolated from cells of the gram-negative bacterium Aquaspirillum itersonii grown under low aeration in the presence of nitrate. The major component, cytochrome c-550, was equated with the (single) c-type cytochrome previously reported to be present in this organism [Clark-Walker, G. D. & Lascelles, J. (1970) Arch. Biochem. Biophys. 136, 153-159], although a significantly higher molecular mass was apparent in the present work. The complete amino acid sequence of this cytochrome is reported in the accompanying paper. A second soluble c-type cytochrome, designated c-556, was also isolated. The molecular mass, isoelectric point, spectrum, midpoint oxidation reduction potential and amino acid composition of this monoheam cytochrome are reported. The possible relationship of this cytochrome to other cytochromes c-556 is discussed. PMID- 3036517 TI - The soluble c-type cytochromes from the bacterium Aquaspirillum itersonii. The complete amino acid sequence of the cytochrome c-550. AB - A complete amino acid sequence is proposed for the cytochrome c-550 isolated from the gram-negative chemo-organotrophic bacterium Aquaspirillum itersonii. The sequence, a single polypeptide chain of 111 residues, was deduced from the sequences of peptides obtained by tryptic, thermolytic or chymotryptic digestion. The cytochrome shows a high degree of sequence homology with the cytochrome c2 from the photosynthetic bacterium Rhodospirillum rubrum, and the evolutionary implications of this are considered. PMID- 3036518 TI - Fundamental relationships for the effect of proton dissociation equilibria on enzymic reaction steps. AB - Relationships have been derived which describe the pH dependence contributed to rate and equilibrium constants for an arbitrary enzymic reaction step by the ionization of an enzymic group or non-enzymic reactant when both protonation states of the ionizing species are assumed to participate in the catalytic reaction. PMID- 3036519 TI - EPR study of ferric native prostaglandin H synthase and its ferrous NO derivative. AB - Purified prostaglandin H synthase (EC 1.14.99.1) apoprotein, a polypeptide of 72 kDA, was titrated with hemin and EPR spectra of high-spin ferric heme were observed at liquid-helium temperature. With up to one hemin per polypeptide, a signal at g = 6.6 and 5.4, rhombicity 7.5%, evolved owing to specifically bound, catalytic active heme. At higher heme/polypeptide ratios signals at g = 6.3 and 5.9 were observed which were assigned to non-specific heme with no catalytic function. In microsomes from ram seminal vesicles the native enzyme showed the signal at g = 6.7 and 5.2 which could not be increased by the addition of hemin. Cyanide, an inhibitor of the enzyme, reacted at lower concentrations with the specific heme abolishing its signal at g = 6.6 and 5.4. Higher concentrations of cyanide were needed for the disappearance of the signal of non-specific heme. The reduced enzyme reacted with NO and formed two types of NO complexes. A transient complex, with a rhombic signal at gx = 2.07, gz = 2.01 and gy = 1.97, was assigned to a six-coordinate complex. The final, stable complex showed an axial signal at g = 2.12 and g = 2.001 and was assigned to a five-coordinate complex, where the protein ligand was no longer bound to the heme iron. Neither type of signal showed a hyperfine splitting from nitrogen of histidine indicating the absence of a histidine-iron bond in the enzyme. From these results and the similarity of the EPR signal at g = 6.6 and 5.4 to the signal of native catalase (EC 1.11.1.6) we speculated that tyrosinate might be the endogenous ligand of the heme in prostaglandin H synthase. PMID- 3036520 TI - Sequential NMR assignments of labile protons in DNA using two-dimensional nuclear Overhauser-enhancement spectroscopy with three jump-and-return pulse sequences. AB - Two-dimensional nuclear Overhauser enhancement (NOESY) spectra of labile protons were recorded in H2O solutions of a protein and of a DNA duplex, using a modification of the standard NOESY experiment with all three 90 degree pulses replaced by jump-and-return sequences. For the protein as well as the DNA fragment the strategically important spectral regions could be recorded with good sensitivity and free of artifacts. Using this procedure, sequence-specific assignments were obtained for the imino protons, C2H of adenine, and C4NH2 of cytosine in a 23-base-pair DNA duplex which includes the 17-base-pair OR3 repressor binding site of bacteriophage lambda. Based on comparison with previously published results on the isolated OR3 binding site, these data were used for a study of chain termination effects on the chemical shifts of imino proton resonances of DNA duplexes. PMID- 3036521 TI - Function of the delipidated beta-adrenergic receptor appears to require a fatty acid or a neutral lipid in addition to phospholipids. AB - Detergent-solubilized preparations of the beta-adrenergic receptor (R) and of the guanyl nucleotide binding proteins (Gs) were extensively treated to remove phospholipids and cholesterol. Reconstitution of an R-Gs system was subsequently performed in the presence of a mixture of natural phosphatidylethanolamine, phosphatidylcholine and phosphatidylserine or the synthetic dioleoyl derivatives of the same phospholipids. In both cases, an additional lipid was required for the agonist-dependent activation of Gs. The requirement could be fulfilled by alpha-tocopherol, or by unsaturated fatty acids such as oleic acid. Inclusion of this non-phosphorylated lipid in the reconstituted system enhanced the isoproterenol-dependent activation of Gs by guanosine 5'-O-[gamma thio]triphosphate 16-33-fold. The rate of activation was largely dependent on the addition of the agonist. Efficient functional reconstitution of R-Gs was thus achieved in a totally defined lipid system. Additional studies of the reconstituted system and of the native membrane led to the notion that the non phosphorylated lipid plays a role in the function of the hormone-R complex. PMID- 3036522 TI - Phosphorylation in vitro of the large subunit of the ribulose-1,5-bisphosphate carboxylase and of the glyceraldehyde-3-phosphate dehydrogenase. AB - A protein kinase activity responsible for the in vitro phosphorylation of at least six endogenous polypeptides including the large subunit of the ribulose-1,5 bisphosphate carboxylase/oxygenase (EC 4.1.1.39) is present in the stroma (3000 X g supernatant, S30) of spinach chloroplasts. The phosphorylation of the ribulose 1,5-bisphosphate carboxylase/oxygenase large subunit is strongly enhanced when sodium fluorure is used as a protein phosphatase inhibitor. Phosphorylation occurs on threonine and serine residues. The protein kinase involved is not Ca2+ dependent. There is also evidence for a protein phosphatase activity which suggests a coupled regulation by a phosphorylation-dephosphorylation process. The phosphorylating activity is drastically reduced when S30 is prepared from leaves harvested after a dark period. Phosphorylation of the ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit is not related to its own synthesis. The in vitro phosphorylation of the glyceraldehyde-3-phosphate dehydrogenase (EC 1.2.1.13) is also demonstrated. PMID- 3036523 TI - Turnover of the phosphomonoester groups of polyphosphoinositol lipids in unstimulated human platelets. AB - The metabolic activity of the polyphosphoinositol lipids in unstimulated human platelets was studied by short-term labelling with [32P]Pi, by replacement of [32P]Pi from pre-labelled platelets with unlabelled phosphate, and by depriving the cells of metabolic ATP. Under short-term labelling conditions, the 4- and 5 phosphate groups of phosphatidylinositol 4-phosphate (PtdIns4P) and phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2] had the same specific 32P radioactivity as the gamma-phosphate of metabolic ATP. The specific 32P radioactivity of the 1-phosphates of phosphatidylinositol, PtdIns4P and PtdIns(4,5)P2 was similar, but only 4-13% compared to that of the ATP-gamma phosphate. When [32P]Pi pre-labelled platelets were incubated with up to 25 mM of unlabelled phosphate, the displacement of the 32P label from PtdIns4P, PtdIns(4,5)P2 and metabolic ATP followed similar kinetics. Inhibition of ATP regeneration in platelets pre-labelled with [32P]Pi resulted in a rapid fall in metabolic ATP with a much slower fall in [32P]PtdIns(4,5)P2, whereas [32P]PtdIns4P increased initially. However, ATP turnover was not abolished, as indicated by the marked (25% of the control) incorporation of extracellular [32P]Pi into PtdIns4P and PtdIns(4,5)P2 in metabolically inhibited platelets. This low phosphate turnover may explain the relative resistance of PtdIns4P and PtdIns(4,5)P2 to metabolic inhibition. We conclude that PtdIns4P and PtdIns(4,5)P2 are present as a single metabolic pool in human platelets. Turnover of the 4- and 5-phosphates of PtdIns4P and PtdIns(4,5)P2 in unstimulated platelets is as rapid as that of the gamma-phosphate of metabolic ATP, and accounts for about 7% of basal ATP consumption. PMID- 3036524 TI - Cloning, nucleotide sequence and expression of the pyrBI operon of Salmonella typhimurium LT2. AB - The pyrB-pyrI region of the Salmonella typhimurium LT2 chromosome has been cloned and sequenced. The two genes were found to constitute an operon, with pyrI being the distal gene and separated from pyrB by a 15-bp intercistronic region. Sequence analysis revealed the presence of two potential promoters; transcription initiated from the promoter proximal to pyrB would produce a transcript which could direct the synthesis of a 33-amino-acid leader peptide. The leader sequence possesses the requisite features of a rho-independent transcriptional terminator (attenuator) which is positioned 22 bp upstream from the pyrB structural gene. A regulatory mutation imparting a 30-fold elevated expression of pyrBI was identified as a two-base-pair deletion in the track of A X T base pairs located on the 3' side of the region of dyad symmetry of the attenuator. The leader sequence also has an additional region of dyad symmetry (putative transcriptional pause site) located 33 nucleotides upstream from the start of the proposed attenuator. The intervening sequence between the putative pause site and the indicated attenuator is characterized by encoding a high content of uracil residues in the transcript. Construction and analysis of transcriptional and translational fusions provided evidence that the leader region has the necessary features to mediate polypeptide synthesis in vivo, the removal of the region corresponding to the pause site and attenuator results in constitutive expression and the more distant potential promoter does not appear to facilitate significant transcriptional activity. Strong homology exists with the pyrBI operon from Escherichia coli K-12 but notable differences are observed. PMID- 3036525 TI - Nucleotide sequence of the chicken proto-oncogene c-erbA corresponding to domain 1 of v-erbA. AB - The nucleotide sequence of the chicken proto-oncogene c-erbA, the cellular counterpart of the viral oncogene v-erbA domain 1, has been determined. The c erbA gene has an exon-intron structure characteristic of the eukaryotic split genes. The c-erbA domain 1 is composed of at least eight exons and seven introns. Analysis of the sequence data reveals a long open reading frame of 239 amino acid residues that share highly significant homology with the viral protein. Our results show that the viral erbA oncogene is a truncated form of its cellular homologue. In fact the cellular gene is 42 nucleotides longer at its 5' coding extremity. The presence of a gag-specific (17/20) nucleotide stretch in this region favours the hypothesis of a homologous recombination event between the cellular erbA and the gag gene of a parental retrovirus. We have also sequenced 1400 bp of DNA lying 5' to the long open reading frame in search of the transcription initiation sites for the c-erbA messenger RNAs. Our findings and interpretations are presented here. PMID- 3036526 TI - Structure and evolutionary origin of the gene encoding mouse NF-M, the middle molecular-mass neurofilament protein. AB - We describe the complete sequence of the gene encoding mouse NF-M, the middle molecular-mass neurofilament protein. The coding sequence is interrupted by two intervening sequences which align perfectly with the first two intervening sequences in the gene encoding NF-L (the low-molecular-mass neurofilament protein); there is no intron in the gene encoding NF-M corresponding to the third intron in NF-L. Therefore, both the number of introns and their arrangement in the genes coding NF-L and NF-M contrast sharply with the number and arrangement of introns in the genes of known sequence, encoding other members of the intermediate filament multigene family (desmin, vimentin, glial fibrillary acidic protein and the acidic and basic keratins); with the exception of a single truncated keratin gene that lacks an encoded tailpiece, these genes all contain eight introns, of which at least six are placed at homologous locations. Assuming the existence of a primordial intermediate filament gene containing most (if not all) the introns found in contemporary non-neurofilament intermediate filament genes, it seems likely that an RNA-mediated transposition event was involved in the generation of an ancestral gene encoding the NF polypeptides. A combination of insertional transposition and gene-duplication events could then explain the anomalous number and placement of introns within these genes. Consistent with this notion, we show that the genes encoding NF-M and NF-L are linked. PMID- 3036527 TI - Influence of the base sequence on the conformational behaviour of DNA polynucleotides in solution. AB - NMR studies were carried out on samples of the non-self-complementary tetramers d(C-A-C-A), d(T-G-T-G), d(G-A-G-A) and d(T-C-T-C) and of 1:1 mixtures of the complementary tetramers d(C-A-C-A) X d(T-G-T-G) and d(G-A-G-A) X d(T-C-T-C) at two DNA concentrations and of the self-complementary octamers d(C-A-C-A-T-G-T-G) and d(G-A-G-A-T-C-T-C). Assignments, based upon one-dimensional NOE and homonuclear-decoupling and two-dimensional correlated and NOE spectroscopies are given of the resonances of most of the base and sugar protons. Chemical shift vs temperature profiles, constructed for all samples, yielded insight into the temperature- and concentration-dependent conformational behaviour of the compounds and were used to obtain thermodynamic parameters pertaining to the stacked-single-strand----random-coil and duplex----random-coil equilibria. Vicinal proton-proton couplings were analyzed in terms of the conformation of the deoxyribose rings in the single-stranded tetramers and duplexed octamers. The NOE patterns, chemical shift profiles, imino-proton resonances and coupling data revealed that the compounds adopt B-DNA-like structures. The ratio duplexed/stacked-single-strand/random coil depends upon external conditions as well as upon base sequence. The thermodynamic data indicate that: in terms of single-helical stacking, the R-R steps (Tm 321-328 K) appear more stable than the Y-R or R-Y steps (Tm 308-316 K) and the Y-Y steps score least (Tm 290-300 K), and the duplexes consisting of alternating, d(Y-R)n, strands are more stable, in terms of delta H degrees, compared to the d(R-R)n X d(Y-Y)n duplexes. The analyses of the couplings demonstrated that the sugars of the single-stranded tetramers and duplexed octamers occur as a blend of N- and S-type conformers, with a preference for the S-type (C2'-endo) sugar conformation: upon duplex formation, no significant shift in the N-type/S-type ratio was observed. The fraction S-type sugar conformation of a given residue, %S, in the stacked-single strands was found to depend upon the nature of its own base and that of the adjacent residues: sugars in an R-R stretch display high values of %S (90-100), whereas those in Y-Y stretches show relatively low values (approximately equal to 65).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3036528 TI - Carcinoma of the urachus demonstrated by CT. AB - Carcinoma of the urachus is a rare tumor with a poor prognosis. Only a few cases explored by CT have been reported until now. This report illustrates the value of CT in establishing the diagnosis and the tumor extension. Furthermore this patient has a probable biological tracer with positive dosage of carcino embryogenic antigen (CEA). PMID- 3036529 TI - Receptor-mediated endocytosis of transferrin and ferritin by guinea-pig reticulocytes. Uptake by a common endocytic pathway. AB - Earlier studies have shown that transferrin binds to specific receptors on the reticulocyte surface, clusters in coated pits and is then internalized via endocytic vesicles. Guinea-pig reticulocytes also have specific receptors for ferritin. In this paper ferritin and transferrin endocytosis by guinea-pig reticulocytes was studied by electron microscopy using the natural electron density of ferritin and colloidal gold-transferrin (AuTf). At 4 degrees C both ligands bound to the cell surface. At 37 degrees C progressive uptake occurred by endocytosis. AuTf and ferritin clustered in the same coated pits and small intracellular vesicles. After 60 min incubations the ligands colocalized to large multivesicular endosomes (MVE), still membrane-bound. MVE subsequently fused with the plasma membrane and released AuTf, ferritin and inclusions by exocytosis. All endocytic structures labelled with AuTf contained ferritin, but 23 to 35% of ferritin-labelled endocytic structures contained no AuTf. These data suggest that ferritin and transferrin are internalized through the same pathway involving receptors, coated pits and vesicles, but that these proteins are recycled only partly in common. PMID- 3036531 TI - Osteomalacia of the mother--rickets of the newborn. AB - During the last 4 years we observed four cases of neonatal rickets. The mothers of the infants suffered from osteomalacia for 1-3 years prior to its diagnosis shortly after the birth of their children. All four infants were born with craniotabes, and one infant had, in addition, a radial fracture. The diagnoses were confirmed by radiological and laboratory tests which revealed a rarefied bone structure, decreased serum 25-hydroxy-vitamin D and increased alkaline phosphatase levels in all patients. The disorder regressed under low-dose vitamin D3 therapy. As osteomalacia seems to be predominant in oriental women living in Berlin, it is necessary to consider vitamin D deficiency when clinical symptoms of this disease arise and to treat these women at least during pregnancy. PMID- 3036532 TI - Intensive chemotherapy in small cell lung cancer. PMID- 3036534 TI - Differences in cleavage of untreated and adriamycin-treated chromatin from normal and leukemic human cells by site non-specific endoribonucleases. PMID- 3036530 TI - The detection of coronary artery disease: a comparison of exercise thallium imaging and exercise equilibrium radionuclide ventriculography. AB - This study compared the accuracy of rest and exercise gated equilibrium technetium ventriculography with exercise thallium imaging in 50 consecutive male patients undergoing routine coronary angiography for the evaluation of chest pain. No patients were excluded on the basis of prior myocardial infarction, nature of angiographically defined coronary disease or symptoms. Antianginal therapy was continued in all patients. Eight patients had normal coronary arteries, 9 had single vessel, disease, 20 had double vessel disease and 13 had triple vessel disease. Sixteen patients had previously documented myocardial infarction. Using exercise radionuclide ventriculography, 34 patients with coronary disease were detected resulting in a sensitivity of 81%; 6 patients with normal coronary arteries had normal scans, a specificity of 75%, with a predictive accuracy of 80%. In comparison, thallium imaging detected 42 patients with coronary disease resulting in a sensitivity of 100%. Six patients with normal coronary arteries had normal thallium images resulting in a specificity of 75% and a predictive accuracy of 96%. These results suggest that exercise thallium imaging is a more accurate investigation than exercise equilibrium radio nuclide ventriculography and is the investigation of choice in the noninvasive detection of coronary artery disease. PMID- 3036533 TI - Fertility in patients with gestational trophoblastic tumors treated with etoposide. AB - The effect of etoposide containing drug combinations on reproductive performance was studied in women successfully treated for gestational trophoblastic disease between 1977 and 1984. Of the 74 women who wished to get pregnant 57 (77.0%) succeeded in having at least one live birth while five (6.8%) have ongoing pregnancies. Of eight (19.8%) women who have not become pregnant two (2.7%) had previous infertility problems antedating the development of gestational trophoblastic disease. The miscarriage rate was 124/1000. There was only one case of foetal death and one congenital abnormality (microcephaly possibly associated with cytomegalovirus infection) among 79 live births and stillbirths. This study indicates that etoposide (VP16-213) as is currently used in this unit for the treatment of gestational trophoblastic tumour is unlikely to have any long-term effect on fertility in most women. PMID- 3036535 TI - TNO6 in non small cell lung cancer. PMID- 3036536 TI - In vitro chemosensitivity of human lung cancer for vindesine. AB - Firstly, the effect of Vindesine was studied on four different human lung carcinoma cell lines (two small cell, one adeno and one squamous cell) with the Fast Green dye exclusion assay (FGA) and the clonogenic assay. Both methods demonstrate a clear dose response relationship and the estimated drug efficacy is similar for both assays. In the cell lines with the longest doubling time a plateau was reached in the FGA, most probably due to the short culture time in this assay. Secondly, the effect of Vindesine on human lung carcinoma specimens (n = 64), mainly bronchoscopy biopsies (n = 48), was evaluated with the FGA. The FGA has merits as predictive test in the clinic in the situation that only a small number of cells can be obtained. In this study, due to the high number of bronchoscopy biopsies only in a minority of cases a conclusion could be obtained (37.5%). PMID- 3036537 TI - Bovine leukemia virus replicates in sheep B lymphocytes under a T cell released factor. AB - In order to determine which cell supports BLV replication in experimentally infected sheep, peripheral blood lymphocytes (PBL) were separated into purified B and T populations by a panning technique. Our data demonstrate that viral replication takes place only in B lymphocytes. However, PHA, a T cell mitogen, is necessary for BLV replication both in PBL and enriched surface immunoglobulin bearing cells, whereas B cell mitogens have no effect on viral replication. Altogether, these results suggest that BLV activation in enriched B lymphocytes is dependent on the presence of residual T cells, and occurs through a T cell interaction, probably mediated by a soluble factor. This possibility was confirmed by the fact that the conditioned medium from cultures of BLV-free sheep T lymphocytes greatly enhances viral production by infected B lymphocytes. Our data favor the hypothesis that BLV multiplication occurs through the regular activation mechanisms of the immune system. PMID- 3036538 TI - Expression and methylation of the Blym gene in human tumor cell lines. AB - We have examined Blym expression in 11 human tumor cell lines. Increased Blym expression was observed in one of three osteosarcoma cell lines relative to nontransformed human foreskin fibroblasts. In addition, enhanced Blym expression was observed in a melanoma cell line and in 2 of 6 squamous carcinoma cell lines relative to nontransformed, low passage human epithelial cells. We found no evidence of gene amplification or rearrangements of Blym sequences in any of the cell lines we have examined. We further analyzed the state of methylation of the Blym gene in several of the tumor cell lines by Msp I/Hpa 11 restriction endonuclease digestion. All cell lines examined had similar Msp I digestion patterns. However, the different tumor cell lines had different Hpa 11 digestion patterns. Therefore, our results indicate that the Blym gene is differentially expressed and methylated in human tumor cell lines. PMID- 3036540 TI - A novel 90-kDa polypeptide (Tp90) possibly involved in an antigen-independent pathway of T cell activation. AB - A novel surface molecule, Tp90, is described which appears to be involved in an antigen-independent pathway of human T lymphocyte activation. The Tp90 molecule was identified by a monoclonal antibody (mAb), MX20, obtained from a fusion using spleen cells of a mouse immunized with cells from two T cell leukemia lines, Jurkat and HPB-ALL. Biochemical data show that Tp90 is distinct and physically independent from the structures already known to be involved in T cell activation, namely T11, T44 or T3/TCR. These results were confirmed by antibody induced antigen modulation experiments. Modulation of Tp90 had no effect on the expression of T3 and of the T cell receptor. Conversely, the expression of Tp90 was not affected by modulation of the T3/TCR molecular complex by either anti-T3 or anti-TCR antibody. Functional studies showed that anti-Tp90 mAb MX20 induced high levels of interleukin 2 production in Jurkat cells. Modulation of the T3/TCR complex significantly decreased the response of Jurkat cells to stimulation by antibody MX20, suggesting that the T3/TCR complex regulates the ability of the Tp90 molecule to induce IL 2 synthesis. In addition to its effect on Jurkat cells, anti-Tp90 mAb was found to be mitogenic for peripheral blood T cells. As the magnitude of the proliferative response elicited by anti-Tp90 mAb was lower than that induced by anti-T3 mAb, the possibility was considered that only a subpopulation of T cells is reactive with anti-Tp90. Indeed as determined by FACS analyses, only 3-14% of E-rosette-positive cells were stained with mAb MX20. In addition, multicolor flow cytometry analysis showed that the Tp90+ cells belong preferentially to the CD8 subset. PMID- 3036539 TI - Activation of B lymphocytes by Epstein-Barr virus/CR2 receptor interaction. AB - Epstein-Barr virus (EBV) infects and transforms human B lymphocytes. The virus receptor was shown to be identical to the complement receptor CR2. The consequences of EBV/CR2 receptor interaction on B lymphocyte activation were analyzed by infection of B cells with the transforming (B95-8) and the nontransforming (P3HR1) virus strains and with UV-inactivated B95-8 virus. Similar to mitogens and antibodies against surface IgM and CR2 receptor, transforming and nontransforming EBV induced the release of leukocyte migration inhibitory factor (LIF). LIF production occurred early after infection and was not affected by irradiation of the B95-8 virus with doses of UV-light which prevented the expression of viral functions. B lymphocytes infected with UV inactivated virus responded to T cell-derived B cell growth factors. The results demonstrate that binding of EBV to the CR2 receptor activates the B cells. Progression of the infected cells in the activation pathway required helper cell derived factors or signals provided by the intact viral genome. The nontransforming P3HR1 virus induced a low level of RNA synthesis and increase of cell size and major histocompatibility complex class I antigen expression. Only the transforming virus induced expression of EBV nuclear antigens and cellular DNA synthesis. PMID- 3036541 TI - Subconvulsive doses of intracisternal bicuculline methiodide, a GABAA receptor antagonist, produce potent analgesia as measured in the tail pinch test in mice. AB - Bicuculline methiodide, a GABAA antagonist produced potent analgesia in the tail pinch test when it was given intracisternally (i.c.) but not intrathecally (i.t.). The ED50 was 5 ng/mouse. This analgesia was antagonized by i.c. muscimol, a GABAA agonist. On the contrary, muscimol (i.t.) produced bicuculline-reversible analgesia. These findings suggest that brain GABA may transmit the nociceptive information while GABA in the spinal cord may inhibit it. PMID- 3036542 TI - Molecular target size of NMDA antagonist binding sites. PMID- 3036543 TI - Direct actions of gossypol on cardiac muscle. AB - Gossypol, an orally active male contraceptive, has been reported to produce cardiotoxicity. In the present study, direct actions of gossypol on the heart muscle were examined using atrial muscle preparations isolated from guinea-pig heart. Gossypol (20-30 microM) produced a transient positive chronotropic effect in spontaneously beating right atrial muscle preparations and a transient positive inotropic effect which reached a peak at about 5 min followed by a marked negative inotropic effect in paced left atrial muscle. These effects of gossypol were not affected by blocking concentrations of phentolamine, nadolol or propranolol, cimetidine and atropine. Gossypol inhibited Na+,K+-ATPase isolated from guinea-pig brain, Na+ pump activity in electrically stimulated left atrial muscle preparations of guinea-pig heart and Na+/Ca2+ exchange in Na+-loaded sarcolemmal vesicles obtained from dog heart. Inhibition of Na+/Ca2+ exchange required high concentrations of gossypol and was rapidly reversible whereas Na+,K+-ATPase inhibition was apparently irreversible. These results indicate that micromolar concentrations of gossypol have direct actions on cardiac muscle. PMID- 3036545 TI - The topography of cholinergic transmission in the mechanism of drug action at muscarinic synapses of the guinea-pig ileum. AB - The pharmacology of cholinergic neurogenic responses evoked by the participation of only the endings of axon terminals was compared to that of responses evoked by participation of the more proximal parts of the terminals also. Myenteric plexus longitudinal muscle strips of the guinea-pig ileum were drawn through narrow orifices in 2 rubber membranes dividing a bath into 3 separate compartments. Oral segments were stimulated electrically by single impulses or by trains and local neurogenic contractions were evoked. The contractions of the aboral segment due to nerve impulses transmitted from the oral segment via the middle segment were also recorded. The opioid ligands ketocyclazocine and [D-Ala2,MePhe4,Met(O)5 ol]enkephalin and noradrenaline inhibited the twitches of the aboral segment evoked by oral segment stimulation more than the local twitches of the oral segment when these agents were applied directly to the respective compartments. The twitches of the aboral segment were also inhibited by the application of these drugs into the middle compartment adjusted to 10 mm width. Verapamil and the alkaline earth metal ions cobalt and lanthanum had similar effects. 4 Aminopyridine increased twitch amplitude more in the aboral segment than in the oral segment when applied directly; similar effects in the aboral segment were seen when the agents were applied to the middle compartment. The action of atropine, papaverine, d-tubocurarine and prostigmine did not discriminate between twitches in the oral and aboral segment when applied directly and all drugs except prostigmine were without effect when applied to the middle compartment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3036544 TI - Evidence that antagonism at non-NMDA receptors results in anticonvulsant action. AB - The effect of gamma-D-glutamylaminomethylsulphonate (gamma-D-GAMS) and 1-(p bromobenzoyl)-piperazine-2,3-dicarboxylate (pBB-PzDA) on convulsions elicited by intracerebroventricular application of kainate (KA) and N-methyl-D-aspartate (NMDA) was studied in mice. gamma-D-GAMS, 0.0025-1.0 mumol, and pBB-PzDA, 0.001 0.2 mumol, were preferentially active against myoclonic seizures induced by kainate, but had also pronounced anticonvulsant action against NMDA. Although pBB PzDA was a more potent anticonvulsant relative to gamma-D-GAMS, gamma-D-GAMS displayed higher kainate-selectivity. gamma-D-GAMS, 0.025 and 0.5 mumol, and pBB PzDA, 0.1 mumol, blocked myoclonic seizures induced by kainate in the presence of 2-amino-7-phosphonoheptanoate, a selective antagonist at the NMDA receptor, with potency comparable to that for antagonism of seizures produced by kainate alone. These results indicate that antagonism at kainate receptors may contribute to anticonvulsant drug action. PMID- 3036547 TI - Aging and platelet beta-adrenoceptor function. AB - Ten young volunteers (mean age 20 years) and ten elderly volunteers (mean age 89 years), including equal numbers of healthy men and women, were tested regarding adrenaline-induced platelet aggregation in vitro, platelet cyclic AMP content before and after beta-adrenoceptor stimulation with isoprenaline, as well as binding by platelet membranes of 125I-hydroxybenzylpindolol. In the aged patients there was a highly significant decrease in the concentration of adrenaline needed to produce irreversible platelet aggregation, as compared with the young. Platelet basal cyclic AMP content did not differ but the response to isoprenaline stimulation, expressed as the percentage rise of cyclic AMP content above the unstimulated level, was significantly decreased in the old subjects. The beta adrenoceptor number was unchanged but the affinity decreased significantly in the old group. The data suggest that the increased aggregation response to adrenaline in old people could be due in part to a diminished functional capacity of the platelet beta-adrenoceptor. PMID- 3036546 TI - Diminished beta-adrenoceptor-mediated relaxation of arteries from spontaneously hypertensive rats before and during development of hypertension. AB - beta-Adrenoceptor agonists and other drugs were studied for their relaxant effects on femoral and mesenteric arterial strips from spontaneously hypertensive rats (SHR). The potency and efficacy of isoproterenol (ISO) in these arteries were decreased in SHR before and during the development of hypertension as compared with age-matched Wistar Kyoto rats (WKY). Reserpine and 6 hydroxydopamine inhibited the development of hypertension but did not alter the reduced ISO-induced relaxation of the arteries. These arteries from prehypertensive SHR (PHSHR) were less sensitive to salbutamol and cyclic AMP and cyclic GMP derivatives than arteries from age-matched WKY. The relaxation response to nitroprusside was less in the femoral but not in the mesenteric arteries from PHSHR than in arteries from age-matched WKY. The relaxation response to papaverine was not diminished in the PHSHR arteries. It was found that the SHR arteries had a reduced responsiveness to the beta-adrenoceptor agonists before the initiation of hypertension and that the diminished relaxation was not specific to the beta-agonists, although there was no generalized defect in vasorelaxation in PHSHR. PMID- 3036548 TI - [3H]PCP-3-OH and (+)[3H]SKF 10047 binding sites in rat brain membranes: evidence of multiplicity. AB - Specific binding of one of the most potent analogs of phencyclidine (PCP), [3H]PCP-3-OH, in rat brain membranes revealed the labeling of high (Kd = 0.5 nM) and low (Kd = 16 nM) affinity binding sites. (+)SKF 10047 potently inhibited high, but not low, affinity [3H]PCP-3-OH binding. (+)[3H]SKF 10047 apparently labeled the high affinity PCP-3-OH binding site and also an additional site, sensitive to haloperidol, which is distinct from the two sites labeled by [3H]PCP 3-OH. PMID- 3036549 TI - The role of cAMP and calcium in the stimulation of proliferation of immature erythroblasts by erythropoietin. AB - The hypothesis that cAMP or calcium are the second messengers of erythropoietin (Epo) was tested on fractionated, Epo-responsive immature erythroblasts from anemic rabbit bone marrow by examining whether the proliferative effects of the hormone could be mimicked by agents that increase the intracellular concentration of cAMP or Ca2+. None of the compounds tested (including 10(-6)-10(-4) M db-cAMP, forskolin, isoprenaline or 10(-7)-10(-6) M of the calcium ionophore A23187) alone or in combination could either initiate or potentiate the mitogenic action of the hormone. Furthermore, addition of 0.2 U/ml erythropoietin produced no permanent or transient increase in the uptake of 45Ca2+ by erythroblasts at 37 degrees C. However, cells cultured with imidazole or cordycepin (which reduce the level of intracellular cAMP), or with the calcium chelator EGTA, or the drugs verapamil or TMB-8 (which interfere with the utilization of extracellular or intracellular calcium) showed a decreased stimulation of DNA synthesis by Epo. Finally, the tumour promoter phorbol ester TPA could partially mimic the action of Epo when added to cultures containing more immature progenitor cells. We conclude then that an artificial increase in the cytoplasmic concentration of either cAMP or Ca2+ is not sufficient to elicit the proliferation of Epo-responsive cells. PMID- 3036551 TI - Fusion of native Sendai virions with human erythrocytes. Quantitation by fluorescence photobleaching recovery. AB - We have recently developed a method to quantitate the fusion of reconstituted viral envelopes with cells by fluorescence photobleaching recovery (FPR) (Aroeti, B & Henis, Y I, Biochemistry 25 (1986) 4588). The method is based on the incorporation of non quenching concentrations of the fluorescent lipid probe N-(7 nitrobenz-2-oxa-1, 3-diazol-4-yl)phosphatidylethanolamine during the reconstitution of the viral envelopes (the latter probe does not incorporate efficiently into the membrane of native virions). In the present work, we employed the fluorescent dye octadecyl rhodamine B chloride (R18), which can be incorporated directly into the membrane of native enveloped virions, to extend the FPR method to study fusion between native Sendai virions and intact human erythrocytes. The R18 fluorescence was found to be quenched in the viral envelope at the concentration range required for the FPR experiments, possibly due to preferential insertion of the probe into specific domains in the viral membrane. We therefore developed a correction (presented in the Appendix) which takes into account the lower quantum yield of the probe molecules in the membranes of unfused virions in the calculation of the fraction of fused virions from the FPR experiments. The results demonstrate that the method does indeed measure virus cell fusion, and that the contribution of exchange to the measurements is not significant. The applicability of the method was further verified by the similarity of the results to those obtained independently by fluorescence dequenching measurements, and its ability to measure the distribution of virus cell fusion within the cell population was demonstrated. These results suggest that the use of R18 can enlarge the scope of the FPR experiments to study the fusion of native virions with cells. PMID- 3036550 TI - Specific modulation of surface receptors in J.774 macrophages by anchorage. AB - The J.774 murine macrophage cells were cultured in suspension in Teflon flasks. When allowed to attach on culture plastic dishes, a 2-3-fold increase in transferrin binding was observed. This occurred in 10 min, reached a steady state at 60 min, remained stable for several hours and was reversible after resuspension of the cells at 37 degrees C. The phenomenon was not dependent on the synthesis of new protein. An opposite change of acetyl LDL receptors was observed, with a threefold decrease of the binding 1 h after the attachment of the cells. The increase of transferrin binding affected almost equally the cell surface and the intracellular sites; therefore it could not be related to a simple shift between these two compartments. It is suggested that the attachment of the cells induced a recruitment of binding sites from a 'silent' pool of receptors. Serum factors, as well as phorbol esters and db-cAMP potentiated the effect of anchorage. PMID- 3036552 TI - Detection of DNA damage in individual cells by flow cytometric analysis using anti-DNA monoclonal antibody. AB - A new method for the measurement of DNA damage in individual cells treated with alkylating agents is described. The method is based on the binding of anti-DNA monoclonal antibody to DNA in situ. Monoclonal antibody F7-26 was obtained by fusion of mouse myeloma cells with spleen cells isolated from a mouse immunized with DNA treated by nitrogen mustard (HN2). Binding of antibody was evaluated by flow cytometry with indirect immunofluorescence. No binding of antibody to DNA in non-treated HeLa S3 cells was detected. Treatment of cells with HN2 or L phenylalanine mustard induced binding of antibody to DNA in situ. Binding of antibody was observed after treating cells with doses of drugs which reduced the surviving fraction below 20%. Intensity of binding increased in proportion to the drug dose. Two-parameter analysis for the antibody binding and DNA content showed no binding of antibody to replicating DNA in control cells. In HN2-treated cells a cell subset with the lowest antibody binding was observed among cells in G1 phase. Binding of antibody to DNA in HN2-treated cells was eliminated by single strand (ss) specific S1 nuclease. In competition assay, antibody was inhibited by thermally denatured DNA, but not by native double-stranded (ds) DNA, RNA, nucleosides and deoxyribohomopolymers. Binding of monoclonal antibody specific for the determinants expressed on ssDNA to the cells treated with alkylating agents may be attributed to local DNA denaturation. Potentiation of L phenylalanine mustard cytotoxicity by buthionine sulfoximine or hyperthermia was accompanied by increased antibody binding to cellular DNA. Immunoreactivity of cells with the monoclonal antibody F7-26 may be a useful probe for the assessment of cell damage induced by alkylating agents, especially in heterogeneous cell populations. PMID- 3036553 TI - Formation and activity of covalent conjugates of poliovirus and ligands binding to cell surface structures. AB - Disulfide-linked conjugates of poliovirus with streptavidin or concanavalin A were formed and the binding of the conjugates to mouse L cells that lack natural poliovirus receptors was studied. The conjugate with streptavidin was specifically bound to biotinylated L cells, but not to unmodified L cells. The conjugate with conA was bound to L cells in the absence of, but not in the presence of alpha-methyl mannoside. Incubation of L cells with bound conjugates did not produce virus, although the conjugates were highly infectious in HeLa cells, containing natural poliovirus receptors. This suggests that the artificially bound virus was unable to penetrate the L cells and start replication. The possibility that binding of the virus to the natural receptor is required for efficient infection is discussed. PMID- 3036554 TI - Studies on heat inactivation of hepatitis A virus with special reference to shellfish. Part 1. Procedures for infection and recovery of virus from laboratory maintained cockles. AB - The consumption of bi-valve molluscan shellfish has been associated with outbreaks of viral gastroenteritis and hepatitis A. Investigations were undertaken to determine the heat inactivation conditions necessary to render shellfish such as cockles safe for the consumer. Conditions for the laboratory maintenance of live cockles are described. In preliminary experiments either poliovirus (10(6) TCID50/ml seawater) or hepatitis A virus (HAV) (approx. 10(4) RFU/ml seawater) was introduced into the shellfish tank. Following 48 h filter feeding, virus was recovered from cockles using an adsorption-elution extraction procedure. Titres of virus recovered ranged from 10(4) to 10(5) TCID50/ml of shellfish extract for poliovirus and from 10(3) to 10(5) RFU/ml of shellfish extract for HAV. Active ingestion of the virus from the seawater was demonstrated by recovering virus from within cockle guts. To quantify recovered HAV, end-point dilutions and an adaptation of a radioimmunofocus assay (RIFA) were compared. The tests were of similar sensitivity but the RIFA has the advantage of being relatively rapid, shortening the time taken to complete an experiment by as much as 4 weeks. PMID- 3036555 TI - Adenosine 3',5'-cyclic-monophosphate and outflow facility in monkey eyes with intact and retrodisplaced ciliary muscle. AB - Bolus intracameral infusion of 100 micrograms cAMP, corresponding to an initial intracameral concentration of 3 mM, significantly increased total outflow facility in cynomolgus monkey eyes with intact or disinserted-retrodisplaced ciliary muscles by 41% and 37% respectively. Lower cAMP doses had little or no effect in both types of eyes, as did dibutyryl cAMP doses of 1-100 micrograms (20 microM-2mM). In eyes with intact ciliary muscles, intracameral infusion (1 and 10 micrograms) of the cAMP analog 8-methylthio cAMP (0.25 mM and 2.5 mM) significantly increased facility by 55% and 69% respectively, while 100 micrograms of the inactive cAMP metabolite 5'AMP (3 mM) did not alter facility. The facility increases produced by cAMP and 8-methylthio cAMP represented a constant percentage of starting facility independent of the absolute value of starting facility. These findings indicate that the facility increase produced by cAMP and appropriate analogs is not mediated by ciliary-muscle contraction and that the disinsertion procedure itself does not compromise the ability of the outflow pathways to respond to cAMP. The facility increase probably represents a cAMP effect directly on the endothelial cells of the trabecular meshwork Schlemm's canal inner wall. PMID- 3036556 TI - Phospholipase C activation via a GTP-binding protein in tumoral islet cells stimulated by carbamylcholine. AB - Carbamylcholine and GTP act synergistically in stimulating the production of [3H]inositol-1-phosphate by digitonized tumoral islet cells (RINm5F line) prelabeled with myo-[2-3H(N)]inositol. The response to these two agents is similar to that evoked by GTP gamma S. These findings suggest that a GTP-binding regulatory protein couples the occupancy of muscarinic receptors to activation of phospholipase C in pancreatic islet cells. PMID- 3036557 TI - Increased responsiveness of the hypothalamic-pituitary axis to steroid feedback effects in ovariectomized rats treated neonatally with monosodium L-glutamate. AB - Chronic ovariectomized rats treated neonatally with MSG showed reduced circulating concentrations of LH coupled with elevated hypothalamic LHRH stores. Despite the apparent loss of LHRH secretion, the small pituitary glands showed an increased density of LHRH receptors and normal responsiveness to the releasing hormone. The positive feedback effects of progesterone on LH release in oestrogen primed animals was greatly exaggerated reflecting the build-up of hypothalamic LHRH stores without loss of pituitary responsiveness to LHRH. PMID- 3036558 TI - Enhancement of fibronectin expression by herbimycin A. AB - Herbimycin A specifically increased the level of fibronectin mRNA in Rous sarcoma virus-infected rat kidney cells, and the time course of fibronectin expression was found to be closely related to that of morphological change induced by herbimycin A. PMID- 3036560 TI - Development of hormone receptors. PMID- 3036561 TI - The mechanism of receptor development as implied by hormonal imprinting studies on unicellular organisms. PMID- 3036562 TI - A new approach to the molecular evolution of hormones: the receptorial aspect. PMID- 3036559 TI - Regulatory peptide receptors: visualization by autoradiography. AB - The receptors for regulatory peptides have been extensively characterized using radioligand binding techniques. By combining these binding techniques with autoradiography it is possible to visualize at the light and electron microscopic levels the anatomical and cellular localization of these receptors. In this review we discuss the procedures used to label peptide receptors for autoradiography and the peculiarities of peptides as ligands. The utilization of autoradiography in mapping peptide receptors in brain and peripheral tissues, some of the new insights revealed by these studies particularly the problem of 'mismatch' between endogenous peptides and receptors, the existence of multiple receptors for a given peptide family and the use of peptide receptor autoradiography in human tissues are also reviewed. PMID- 3036563 TI - Receptors for intercellular messenger molecules in microbes: similarities to vertebrate receptors and possible implications for diseases in man. PMID- 3036564 TI - Development of hormone receptors: conclusion. PMID- 3036565 TI - Internalization of polypeptide hormones and receptor recycling. PMID- 3036567 TI - Receptor ontogeny and hormonal imprinting. PMID- 3036566 TI - Why do hormone receptors arise? An introduction. PMID- 3036569 TI - The existence of B/E and E receptors on Hep-G2 cells: a study using colloidal gold- and 125I-labeled lipoproteins. AB - The presence of specific receptors for apolipoprotein B (low-density lipoproteins) and apolipoprotein E (HDL-E) on Hep-G2 cells and human skin fibroblasts was studied by chemical methods and by electron microscopy using a differential gold labeling technique. Fibroblasts bound both types of lipoproteins to one and the same receptor (B/E receptor) as deduced from competition experiments with HDL-E and LDL. Labeled HDL-E, on the other hand, was only partially displaced by cold LDL but was completely displaced by unlabeled HDL-E. Scatchard analysis of lipoprotein binding to Hep-G2 cells revealed an approx 10 times higher binding affinity of apoE-containing lipoproteins as compared to apoB-containing ones. No differences between apoE- or apoB-containing lipoproteins with respect to the morphology of cell binding and intracellular processing were observed. The results are compatible with the concept that Hep-G2 cells possess two kinds of receptors, one specific for apoB- and apoE-containing lipoproteins (B/E receptor) and another specific for apoE only. From these studies we conclude that Hep-G2 cells may serve as a suitable model for studying the lipoprotein metabolism in the liver. PMID- 3036568 TI - Activation of human blood monocytes by adherence to tissue culture plastic surfaces. AB - Human monocytes, purified by countercurrent centrifugal elutriation, were cultured either in plastic dishes or in Teflon vials to determine if attachment would result in activation. beta-Glucuronidase activity, 5'-nucleotidase activity, plasminogen activator, and superoxide anion generation were measured as markers of monocyte activation. Conditioned media and cell lysates were assayed at 2, 4, 8, and 10 hr and then daily for 6 days. Monocytes cultured in plastic dishes secreted a significantly greater proportion of their beta-glucuronidase into the medium than those cultured in Teflon vials. The activity of 5' nucleotidase was lower in monocytes cultured in plastic dishes, consistent with greater activation. Cellular plasminogen activator levels and the capacity for superoxide anion generation were enhanced in cells cultured in plastic dishes, relative to monocytes cultured in Teflon vials. These observations indicate that monocyte attachment in plastic surfaces results in their activation, a phenomenon that may influence the nature and interpretation of experimental data derived from cultured adherent monocytes or macrophages. PMID- 3036570 TI - Heterogeneity of TdT+, HLA-DR+ acute leukaemia: immunological, immunocytochemical and clinical evidence of lymphoid and myeloid origin. AB - 15 cases of acute leukemia (AL) displaying a TdT+, HLA-DR+ phenotype were studied; surface immunoglobulins, T cell markers and the common acute lymphoblastic leukaemia (c-ALL) antigen were negative, as were peroxidase and non specific esterase cytochemical reactions. All cases were extensively investigated by conventional immunofluorescence (IF) and immunoperoxidase (IP), with a panel of monoclonal antibodies (MoAb), using both light and electron microscopy, and for ultrastructural myeloperoxidase (MPO). 8 cases, which were OKB2+, BA1+, B4+, J5- and BA2- by IF, expressed the J5 antigen in IP. These cases were therefore re classified as ALL with a weak expression of the C-ALL antigen. The other 7 cases showed an OKB2-, BA1-, B4+, BA2+ phenotype at IF and were also positive for 1 or more anti-myeloid MoAb. These features were confirmed by IP study. 4 patients also presented ultrastructural positivity to MPO. These cases were considered as proliferations of early precursor cells capable of expressing both myeloid and lymphoid features. This study, while demonstrating the heterogeneity of TdT+, HLA DR+ AL, suggests that the cell origin of many cases may be defined by extensive immunotyping at both IF and IP level. The prognostic and therapeutic implications of these findings are discussed, also in view of the poor prognosis often observed in the more undifferentiated cases of AL. PMID- 3036571 TI - Diagnostic and prognostic significance of serum measurements of lactoferrin, lysozyme and myeloperoxidase in acute myeloid leukemia (AML): recognition of a new variant, high-lactoferrin AML. AB - 92 patients with acute myeloid leukemia were classified according to the FAB classification (M1 n = 20, M2 n = 43, M3 n = 1, M4 n = 19, M5a n = 2, M5b n = 2, and M6 n = 5 patients). Serum measurements of lactoferrin (LF), myeloperoxidase (MPO) and lysozyme (LYS) were performed before the start of treatment. LF was significantly lower in M1 when compared with M2 but not as compared to M4, MPO was significantly higher in M2 and M4 than in M1, but comparable MPO levels were found in M2 and M4. LYS was significantly elevated in M2 in comparison with M1, and in M4 when compared to both M1 and M2. Polymorphonuclear granulocytes (PMNs) in M1 were significantly reduced when compared with M2 and M4, whereas mononuclear cells were significantly increased in M4 in comparison with both M1 and M2. FAB classification did not generate any prognostic information. When the patients were, instead, subdivided according to LF levels were found prognostically significant differences. Of patients below 100 micrograms/l, 44% went into remission as compared to 77% with LF from 101 to 400 micrograms/l. In patients with LF levels above 400 micrograms/l the remission frequency was only 14%. Multivariate statistical analysis on the data further suggested that lactoferrin may be used as an independent prognostic indicator. We conclude that although determination of the serum-levels of lactoferrin, lysozyme and myeloperoxidase in certain cases may be valuable as a supplement to the morphological examination of acute myeloid leukemia, it is evident that none of the three determinations can be used alone to distinguish between the FAB groups. PMID- 3036572 TI - Affinity modification of E1-form of Na+, K+-ATPase revealed Asp-710 in the catalytic site. AB - An alkylating ATP analogue, gamma-[4-(N-2-chlorethyl-N-methylamino)]benzylamide ATP (C1RATP), covalently binds to the catalytic alpha-subunit of Na+, K+-ATPase yielding a product resistant to hydrolysis by the enzyme and inhibiting the ATP hydrolysing activity. The Na+-form of the membrane-bound Na+, K+-ATPase modified with C1RATP was hydrolysed by pepsin under conditions providing maximum stability of the modification product (4 degrees C, pH 1.5). The modified peptide was isolated by HPLC and its amino acid sequence was found to involve residues 706 713 of the alpha-subunit polypeptide chain. This fragment located near the gamma phosphate of ATP is a component of the active site. It is highly homologous with corresponding regions of the catalytic subunits of all the known E1-E2 ATPases. In the Na+-(or E1-)enzyme form Asp-710 is the target of modification. Evidently E1- and E2-enzymes have different targets in C1RATP modification, i.e. the polypeptide chain regions near the ATP gamma-phosphate in the enzyme active site differ somewhat in their conformations. PMID- 3036573 TI - Does the inositol tris/tetrakisphosphate pathway exist in rat heart? AB - Appearance of two isomers of inositol trisphosphate (InsP3) was observed when [3H]inositol prelabelled rat heart ventricles were stimulated for 10 and 30 s with noradrenaline. In contrast, inositol tetrakisphosphate (InsP4) could not be detected. However the existence of the inositol tris/tetrakisphosphate pathway was demonstrated by studying [3H]inositol 1,4,5-trisphosphate (Ins-1,4,5-P3) metabolism in a soluble fraction of rat heart. There, [3H]Ins-1,4,5-P3 was phosphorylated to form [3H]Ins-1,3,4,5-P4. Raising [Ca2+] from 1 nM to 1 microM increased InsP3 kinase activity by 2-fold (EC50 for Ca2+ approx. 56 nM). This effect appeared to be due to an increase of the apparent Vmax of the enzyme while the apparent Km was unchanged. PMID- 3036574 TI - Chromosomal localization of the gene coding for alpha-subunit of Na+,K+-ATPase in the American mink (Mustela vison). AB - The gene coding for the alpha-subunit of Na+,K+-ATPase has been localized on chromosome 2 of the American mink (Mustela vison) using the somatic cell hybrids mink-Chinese hamster and pig cDNA clones as hybridization probes. PMID- 3036576 TI - Enhanced prostaglandin E2 and thromboxane B2 release from resident peritoneal macrophages isolated from morphine-dependent rats. AB - Resident peritoneal macrophages from morphine-addicted rats (4 days) released more prostaglandin (PG) E2 and thromboxane (Tx) B2, but not 6-keto-PGF1 alpha, than cells from control animals. This effect, which was due to an enhancement of endogenous AA turnover, was not related to any changes in cAMP synthesis or lysosomal enzyme secretion. [D-Ala2]-Met-enkephalin had no effect on eicosanoid release in vitro. Both morphine and PGE2 have been shown to depress macrophage functions. We suggest that morphine-stimulated macrophage PGE2 synthesis, and the consequent inhibition of phagocytosis, could contribute to the decreased resistance to infections associated with opiate addiction. PMID- 3036575 TI - Proposal that the function of the membrane-bound cytochrome a1-like haemoprotein (cytochrome b-595) in Escherichia coli is a direct electron donation to cytochrome d. AB - The cytochrome d-containing oxidase of oxygen-limited Escherichia coli comprises cytochromes d, cytochrome b-558 and cytochrome b-595, previously called cytochrome a1. The reaction of the fully reduced complex with oxygen involves ligand binding to the ferrous haem d to form an oxygenated species, followed by oxidation of two b-type cytochromes, whose identity is unclear. Here we report kinetic studies on cytochrome b-595 oxidation and suggest that these results, together with optical and EPR data on the oxidase complex and its reaction with oxygen, are consistent with the hypothesis that the role of cytochrome b-595 is further reduction of the oxygen bound to cytochrome d. PMID- 3036577 TI - Smooth muscle myosin phosphatase inhibition and force enhancement by black sponge toxin. AB - Smooth muscle contraction depends on the state of myosin phosphorylation and hence on the balance of myosin light chain kinase and phosphatase activity. Effects of okadaic acid isolated from black sponge on both enzyme activities and contractility were studied in chemically skinned fibers from guinea pig taenia coli. The toxin strongly inhibits myosin phosphatase and enhances tension development. PMID- 3036578 TI - Intracellular organelle motility and membrane fusion processes in human neutrophils upon cell activation. AB - Release and subcellular fractionation experiments indicate that fusion of a novel tertiary granule with the plasma membrane is concomitant with human neutrophil activation. Phorbol 12-myristate 13-acetate (PMA) induced a respiratory burst in human neutrophils as well as a high release of gelatinase, a marker of the tertiary granule. Preincubation of neutrophils with cytochalasin E induced a partially activated or 'primed' state, in which cells were unable to generate superoxide anion, but showed a reduced latency period for this activity. Fusion of tertiary granules with the cell surface also occurred during priming, although to a lesser extent than in PMA stimulation. The rapid tertiary granule degranulation, preceding that of specifics and azurophilics, seems to play an important role in the functionality and secretory properties of human neutrophils. PMID- 3036579 TI - Hormonal regulation of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase: kinetic models. AB - A graphical method to reveal the so-called 'critical fragments' in schemes of biochemical systems is considered. These fragments produce multiple steady states or self-oscillations in systems. As an example, the bifunctional enzyme, 6 phosphofructo-2-kinase/fructose-2,6-bisphosphatase, regulated by glucagon through enzyme phosphorylation, is discussed. It is shown that this enzyme may act as a metabolic switching mechanism in discontinuous or oscillatory regimes, depending on the specific structure of its kinetic scheme. The boundaries of concentrational and parameter domains for these critical phenomena are also predicted. PMID- 3036580 TI - Endogenously generated 5-hydroperoxyeicosatetraenoic acid is the preferred substrate for human leukocyte leukotriene A4 synthase activity. AB - A single protein from human leukocytes possesses both 5-lipoxygenase and leukotriene A4 (LTA4) synthase activities. It has been reported that LTA4 production is more efficient when the enzyme utilizes arachidonic acid, than when 5-HPETE is exogenously supplied as substrate. In the present study, human leukocyte homogenate 100,000 X g supernatant was incubated with 100 microM octadeuterated arachidonic acid and exogenous 5-HPETE (0-80 microM), and the isotopic composition of LTA4 hydrolysis products was determined by gas chromatography-mass spectrometry. Even though 100 microM deuterated arachidonic acid results in 20-30 microM deuterated 5-HPETE, 80 microM exogenous 5-HPETE in the incubation could reduce the amount of deuterated LTA4 by only approx. 20%. The present study would thus indicate that the arachidonic acid moiety is preferentially converted to LTA4 in a concerted reaction without dissociation of a 5-HPETE intermediate. PMID- 3036581 TI - Detailed structural analysis of exposed domains of membrane-bound Na+,K+-ATPase. A model of transmembrane arrangement. AB - Exposed regions of the alpha- and beta-subunits of membrane-bound Na+,K+-ATPase were in turn hydrolyzed with trypsin. Resistance of the beta-subunit to proteolysis was shown to be due mainly to the presence of disulfide bridge(s) in the molecule. A model for the spatial organisation of the enzyme in the membrane was proposed on the basis of detailed structural analysis of extramembrane regions of both subunits. PMID- 3036582 TI - The family of human Na+,K+-ATPase genes. No less than five genes and/or pseudogenes related to the alpha-subunit. AB - Five different nucleotide sequences have been found in the human genome homologous to the gene of the alpha-subunit of Na+,K+-ATPase. A comparative analysis of the primary structure of these genes in the region 749-1328 (in coordinates of cDNA from the pig alpha-subunit) is presented. PMID- 3036583 TI - Unimpaired coupling of phosphorylated, desensitized beta-adrenoceptor to Gs in a reconstitution system. AB - Heterologous desensitization of turkey erythrocyte beta-adrenoceptors correlates with receptor phosphorylation and impaired receptor-Gs coupling, as assessed by fusion of purified desensitized receptors with X. laevis erythrocytes [(1984) Science 225, 837-840]. We have purified beta-receptors from desensitized and untreated turkey erythrocytes and have compared the abilities of these two receptors to couple with pure turkey erythrocyte Gs in a reconstituted system. Functional receptor-Gs coupling was assessed by measuring hormone-dependent Gs activation by GTP gamma S and GTPase activity. While in membranes prepared from desensitized cells, receptor-Gs coupling was clearly reduced, this effect was absent when coupling of purified desensitized receptor was measured. We conclude that covalent modification by phosphorylation does not fully explain the functional uncoupling at the membrane level. PMID- 3036584 TI - Activation of metallothionein expression is potentiated by DNA sequences present in the herpes simplex virus thymidine kinase gene. AB - A mouse cell line (Ltk-aprt-) which is resistant to the anti-viral effects of interferon also has a reduced ability to synthesize metallothionein on exposure to cadmium. Like the ability to respond to interferon, cadmium-induced metallothionein synthesis is restored to wild-type levels in clones obtained by introducing a thymidine kinase gene into Ltk-aprt-cells. Transfection of other genes does not have such an effect. Since metallothionein expression is also activated by interferon the results suggest that the regulation of several genes which are responsive to interferon can be modulated by specific sequences present in the Herpes virus thymidine kinase gene. PMID- 3036585 TI - Phosphorylation of type-L pyruvate kinase in intact hepatocytes. Localisation of the phosphorylation site in response to both glucagon and the Ca2+-linked agonist phenylephrine. AB - Pyruvate kinase is one of the enzymes which can be phosphorylated by stimulation of the cell with either glucagon or Ca2+-linked hormones. Whether these two classes of hormones phosphorylate the same site on the enzyme is unclear. Our results demonstrate that isolation of [32P]phosphorylated type-L pyruvate kinase from glucagon-treated hepatocytes followed by aspartyl-prolyl cleavage yields a [32P]phosphorylated peptide of Mr 17,000. This fragment is also phosphorylated in response to the Ca2+-mediated agonist phenylephrine. PMID- 3036586 TI - An enzyme catalyzing the liberation of N-acetylglucosamine from N acetylglucosaminyl pyrophosphorylpolyprenol in Bacillus polymyxa membranes. AB - A novel enzyme which specifically hydrolyzes N-acetylglucosaminyl pyrophosphorylpolyprenol to liberate N-acetylglucosamine was found in membranes of Bacillus polymyxa AHU 1385. The enzyme seems to be inactive toward alpha-N acetylglucosaminyl phosphorylundecaprenol, beta-N-acetylglucosaminyl phosphorylundecaprenol, N-acetylglucosamine 1-phosphate, N-acetylglucosamine 1 pyrophosphate, or UDP-N-acetylglucosamine. Much lower activities of the same enzyme were also found in membranes of several other strains of Bacilli. PMID- 3036587 TI - Stimulation of partially purified adenylate cyclase from bull sperm by bicarbonate. AB - Solubilized and partially purified adenylate cyclase from bull sperm was found to be specifically activated (up to 6-fold) by sodium bicarbonate (NaHCO3) and to a lesser extent by NaNO3. Other sodium salts were either ineffective (e.g. NaCOOH) or inhibitory (e.g. NaHSO3, NaHSO4 and Na2B4O7). Stimulation by NaHCO3 was dose dependent in the range of 0-40 mM and was greater when enzyme activity was assayed in the presence of magnesium as compared with manganese ions. Bicarbonate seems to affect maximal enzyme velocity (Vmax) and has no effect on the Km of adenylate cyclase for Mn-ATP. Stimulation of adenylate cyclase by NaHCO3 coincided with the elution pattern of the enzyme as recorded following chromatography on DEAE-cellulose or gel filtration on BioGel P-100. These results suggest that in the course of stimulation of sperm adenylate cyclase, bicarbonate is likely to interact directly with the enzyme. Furthermore, this intrinsic and unique property of sperm adenylate cyclase may explain results reported by others on the stimulation of cAMP production by bicarbonate in intact and broken sperm preparations and suggest a biochemical basis for enhanced sperm motility associated with high bicarbonate concentrations. PMID- 3036588 TI - Phosphoinositide metabolism in resting and thrombin-stimulated human platelets. Evidence against metabolic heterogeneity. AB - The specific radioactivity of the phosphodiester and phosphomonoester moieties of the phosphoinositides was determined in resting and thrombin-stimulated platelets that were prelabelled with [32P] Pi. In the unstimulated cells, the specific radioactivity of the monoester phosphates of PIP and PIP2 was similar. Both prolonged incubation at 37 degrees C and upon stimulation with 0.5 U/ml of thrombin, the specific radioactivity of the monoester phosphates decreased 10-15% in both polyphosphoinositides. In the unstimulated cells, the specific radioactivity of the diester phosphates was similar in PI, PIP and PIP2 but amounted only to 3% of the activity of the monoester groups. Prolonged incubation of the unstimulated cells as well as stimulation with thrombin induced a similar 5-6 fold increase in specific radioactivity of the diester phosphate of PI, PIP and PIP2. The results indicate that the phosphoinositides in both resting and thrombin-stimulated platelets exist in a metabolically homogeneous pool. PMID- 3036590 TI - Comparative EPR studies on the nitrite reductases from Escherichia coli and Wolinella succinogenes. AB - Hexaheme nitrite reductases purified to homogeneity from Escherichia coli K-12 and Wolinella succinogenes were studied by low-temperature EPR spectroscopy. In their isolated states, the two enzymes revealed nearly identical EPR spectra when measured at 12 K. Both high-spin and low-spin ferric heme EPR resonances with g values of 9.7, 3.7, 2.9, 2.3 and 1.5 were observed. These signals disappeared upon reduction by dithionite. Reaction of reduced enzyme with nitrite resulted in the formation of ferrous heme-NO complexes with distinct EPR spectral characteristics. The heme-NO complexes formed with the two enzymes differed, however, in g values and line-shapes. When reacted with hydroxylamine, reduced enzymes also showed the formation of ferrous heme-NO complexes. These results suggested the involvement of an enzyme-bound NO intermediate during the six electron reduction of nitrite to ammonia catalyzed by these two hexaheme nitrite reductases. Heme proteins that can either expose bound NO to reduction or release it are significant components of both assimilatory and dissimilatory metabolisms of nitrate. The different ferrous heme-NO complexes detected for the two enzymes indicated, nevertheless, their subtle variation in heme reactivity during the reduction reaction. PMID- 3036589 TI - Arginine-vasopressin stimulates the formation of phosphatidic acid in rat Leydig cells. AB - Arginine-vasopressin (AVP) stimulated the formation of labelled phosphatidic acid (PA) in [14C]arachidonic acid-prelabelled rat Leydig cells. After addition of 10( 6)M AVP [14C]arachidonoylphosphatidic acid reached a maximum within 2 min. The increase was dose-dependent (10(-11)-10(-6)M). No change in labelling of other phospholipids and diacylglycerol could be detected. The V1 antagonist dPTyr(Me)AVP inhibited in a dose-dependent manner the AVP-stimulated accumulation of PA. The V2 agonist dPVDAVP was without effect. The present results suggest that AVP binds to V1 receptors in rat Leydig cells resulting in stimulation of PA turnover. We suggest that the AVP-stimulated PA formation is an indication of phosphoinositide turnover. PMID- 3036591 TI - Aspirin induces alterations in low-density lipoprotein and decreases its catabolism by cultured human fibroblasts. AB - Aspirin interacts in vitro with human low-density lipoprotein (LDL), which results in a decrease in free amino groups of apolipoprotein B and an increase of electrophoretic mobility of the particle. The aspirin-treated LDL was less efficiently recognized than native LDL by the apo B/E receptor of fibroblasts. These results suggest that aspirin in long-term treatment could influence the LDL receptor pathway. However, aspirin-treated LDL did not bind to the scavenger receptor of macrophages. PMID- 3036592 TI - Molecular cloning of the myelin specific enzyme 2',3'-cyclic-nucleotide 3' phosphohydrolase. AB - We describe the isolation of cDNA clones for bovine brain 2',3'-cyclic-nucleotide 3'-phosphohydrolase (CNPase, EC 3.1.4.37), the third most abundant protein in central nervous system myelin. The cDNA encodes the complete protein (400 amino acids) and hybridizes to a major size species of mRNA in bovine brain tissue, approx. 2.7 kb in size. CNPase mRNA levels do not appear to be affected in quaking dysmyelinating mutant mice. The sequence reveals probable sites for CNPase phosphorylation by cAMP-dependent protein kinase and a region of homology with haemocyanin. PMID- 3036594 TI - The mechanism of action of aspartic proteases involves 'push-pull' catalysis. AB - In accord with the available kinetic and X-ray crystallographic data, it is proposed that the two catalytically competent carboxyl groups of aspartic proteases constitute a functional unit which mediates the proton from the attacking water molecule to the leaving nitrogen atom of the substrate. Protonation of this nitrogen atom has been the main issue of the previous mechanistic proposals. The first step of the present mechanism involves proton transfer from the water to the aspartic diad and concurrently another proton transfer from the diad to the carbonyl oxygen of the scissile peptide bond. These proton transfers provide the driving force for the bond formation between the substrate and water, which leads to the formation of a tetrahedral intermediate. The intermediate breaks down to products by a similar facilitation, i.e. by concerted general acid-base catalysis, which involves simultaneous proton transfers from the intermediate to the diad and from the diad to the leaving nitrogen of the substrate. The symmetrical mechanism of the formation and decomposition of the tetrahedral adduct resembles that found in the serine protease catalysis. PMID- 3036593 TI - Purification of all thirteen polypeptides of bovine heart cytochrome c oxidase from one aliquot of enzyme. Characterization of bovine fetal heart cytochrome c oxidase. AB - A protocol has been worked out for separating all thirteen different polypeptides in the beef heart cytochrome c oxidase complex from a single aliquot of enzyme. This involves an initial separation of polypeptides by gel filtration on a Biogel P-60 column in SDS, a step which purifies subunits CIV and CVIII and gives mixtures of CV + CVI, ASA, AED and STA, as well as CVII, CIX and IHQ. These mixtures are then resolved by reverse-phase high-performance liquid chromatography. The separation procedures have been applied to fetal heart cytochrome c oxidase of gestation between 100 and 200 days. No differences were found in the N-terminal sequences of any of the cytoplasmically made subunits or in the entire sequence of CIX between late fetal and adult forms of the enzyme. PMID- 3036595 TI - Catecholamine inhibition of Ca2+-induced insulin secretion from electrically permeabilised islets of Langerhans. AB - Noradrenaline (1-10 microM) inhibited Ca2+-induced insulin secretion from electrically permeabilised islets of Langerhans with an efficacy similar to that for inhibition of glucose-induced insulin secretion from intact islets. The inhibition of insulin secretion from permeabilised islets was blocked by the alpha 2-adrenoreceptor antagonist, yohimbine. Adenosine 3',5'-cyclic monophosphate (cAMP) did not relieve the noradrenaline inhibition of Ca2+-induced secretion from the permeabilised islets, although noradrenaline did not affect the secretory responses to cAMP at substimulatory (50 nM) concentrations of Ca2+. These results suggest that catecholamines do not inhibit insulin secretion solely by reducing B-cell adenylate cyclase activity, and imply that one site of action of noradrenaline is at a late stage in the secretory process. PMID- 3036596 TI - Isolation and amino acid sequence of the 'Rieske' iron sulfur protein of beef heart ubiquinol:cytochrome c reductase. AB - The sequence of the 'Rieske' iron sulfur protein from the bc1 complex of beef heart mitochondria has been determined by solid phase Edman degradation of the whole protein and of various proteolytic fragments. The protein consists of 196 amino acid residues. The molecular mass of the apoprotein was calculated to be 21,536 Da, that of the holo-protein including the Fe2S2 cluster as 21,708 Da. The protein is mainly hydrophilic with a polarity index of 42.9% and 25% of charged residues. It contains a hydrophobic membrane anchor which is predicted to form a 'hairpin' structure. The iron sulfur cluster is bound near the C-terminus of the protein between a hydrophobic and a more amphipathic domain. This reflects the fact that the cluster is located near the outer surface of the inner mitochondrial membrane. A folding pattern describing all known features of the protein is proposed. PMID- 3036597 TI - One-electron reduction of an anthracycline antibiotic carminomycin by a human erythrocyte redox chain. AB - Human erythrocyte membranes catalyse the NAD(P)H-dependent generation of the semiquinone of an adriamycin-type antibiotic carminomycin under anaerobic conditions. The maximal yield of the antibiotic radical is about 4-fold higher in the presence of NADPH than of NADH. The possible significance of the antibiotic reduction to the semiquinone by a human erythrocyte membrane redox chain for the clinical usage of these antibiotics is discussed. PMID- 3036598 TI - Ultrasonic studies of proton-transfer reactions at the catalytic site of alpha chymotrypsin. AB - Ultrasonic relaxation measurements for alpha-chymotrypsin in phosphate, sulfite and arsenate buffers exhibit a high peak of absorption at neutral pH. The analysis is based on: comparison of the relaxation measurements for the enzyme and for the zymogen and inhibited enzyme; X-ray and neutron diffraction data, and high-resolution NMR data. The ultrasonic relaxation is shown to result mainly from a proton-transfer reaction that involves the histidine at the catalytic site (His-57). The question is raised of whether the enhanced ultrasonic effect observed in the enzyme is indicative of a property that plays a part in the catalytic activity. PMID- 3036599 TI - Electron paramagnetic resonance and magnetic circular dichroism studies of a hexa heme nitrite reductase from Wolinella succinogenes. AB - The nature of the heme centers in the hexa-heme dissimilatory nitrite reductase from the bacterium Wolinella succinogenes has been investigated with EPR and magnetic circular dichroism spectroscopy. The EPR spectrum of the ferric enzyme is complex showing, in addition to magnetically isolated low-spin ferric hemes with g values of 2.93, 2.3 and 1.48, two sets of signals at g = 10.3, 3.7 and 4.8, 3.21, which we assign to two pairs of exchange coupled hemes. The MCD spectra show that the isolated hemes are bis-histidine coordinated and that there is one high-spin ferric heme. The exchange coupling is lost on treatment with SDS. PMID- 3036600 TI - A TY1 element is inserted in the CYR1 control region of Saccharomyces cerevisiae strain AB320. AB - Southern blotting using a 5'-proximal probe of the Saccharomyces cerevisiae CYR1 gene has revealed heterogeneity in laboratory strains. It is demonstrated that strain AB320 contains a Ty1 element inserted in the promoter region of CYR1. The Ty1 orientation suggests that transcription of CYR1 is initiated downstream from the insertion region. PMID- 3036601 TI - Dexamethasone-dependent expression of beta 1-24 corticotropin stimulated adenylate cyclase during adipose conversion of 3T3-F442A cells. AB - When 3T3-F442A preadipocytes were grown in culture media supplemented with corticosteroid poor fetal calf serum and insulin they differentiated into adipocytes. Glycerophosphate dehydrogenase, a marker of terminal differentiation, developed a 600-fold increase of activity whereas the adenylate cyclase system remained unresponsive to the synthetic ACTH(1-24) analog. In contrast, 3T3-F442A adipocytes, differentiated in the presence of dexamethasone, exhibited an adenylate cyclase activity which was stimulated 4-fold by ACTH(1-24). The stimulation of the adenylate cyclase activity by GTP gamma S remained unchanged (about 20-25-fold) suggesting that the G regulatory coupling protein was not functionally modified by dexamethasone. Binding studies with 125I-ACTH revealed that specific cellular binding could be evidenced in dexamethasone-treated cells while control adipocytes did not exhibit any specific binding of 125I-ACTH. These findings lend support to the hypothesis that the setting off of this ACTH responsiveness in 3T3-F442A cells is regulated by dexamethasone after cells are committed to adipose differentiation. PMID- 3036602 TI - Light- and nucleotide-dependent increase in apparent viscosity in a suspension of retinal disks. AB - Light triggers the cyclic nucleotide cascade in photoreceptor disk membranes. We report here that light-induced changes in the apparent viscosity of disk membrane suspensions can also be observed using either native disk membranes or washed membranes reconstituted with G protein and PDE. The viscosity changes are light- and GTP-dependent and require the presence of G protein and PDE. The magnitude of the viscosity change increases with increasing membrane concentration. Under the same conditions in which light elicits a change in viscosity, we observe a large increase in light scattering by the disk membrane suspension. PMID- 3036603 TI - Poorly differentiated human gastric carcinoma is more sensitive to antitumor drugs than is well differentiated carcinoma. AB - The chemosensitivities of 41 poorly differentiated gastric cancer tissues were compared with that of 16 well differentiated tissues, using the in vitro succinate dehydrogenase inhibition test. These human tissues obtained at the time of surgery were exposed to six different antitumor drugs: carboquone (CQ), adriamycin (ADM), mitomycin C (MMC), aclacinomycin A (ACR), cisplatin (DDP) and 5 fluorouracil (5-FU). The chemosensitivity was determined as positive when the succinate dehydrogenase (SD) activity of the drug exposed cells was decreased to below 50% of that of control cells, on day 3 of exposure. Decrease in SD activity was remarkable in the poorly differentiated tissues, compared to the well differentiated tissues, exposed to ADM, MMC, DDP and 5-FU. The sensitive rates were higher in the poorly differentiated tissues than in the well differentiated tissues, against all six antitumor drugs. Sixty-three per cent of the poorly differentiated tissues were sensitive to more than three antitumor drugs, in an identical tissue, but the rate was only 19% in the well differentiated tissues. The resistant rates to all drugs tested were 20% in the poorly differentiated and 31% in the well differentiated tissues. This would indicate that patients with a poorly differentiated gastric cancer will probably show a better response to antitumor drugs, compared to those with a well differentiated type. PMID- 3036604 TI - Chromosome changes of in situ carcinomas in the female breast. AB - Eight lesions of ductal carcinoma in situ (DCIS) and one case of benign radial scar were investigated cytogenetically and compared with invasive ductal carcinomas. DCIS was shown to be associated with chromosome changes different from those of invasive carcinomas as concern ploidy levels. All cases exclusively contained abnormal metaphases, also the specimen that was classified as a benign radial scar. No consistent chromosome changes were found. Chromosome no. 1 was most frequently involved in marker chromosome formation. PMID- 3036606 TI - Cardioselective muscarinic antagonists. PMID- 3036605 TI - Complexity and significance in computer simulations of physiological systems. AB - Complexity in a theoretical model may or may not be associated with a high level of arbitrariness, depending on how the model is constructed. In this paper I use examples from cardiac electrophysiology to illustrate two techniques for maintaining significance in complex simulations. First, the approaches of analysis and synthesis are compared as methods of constructing models of complex systems. When a model is constructed by synthesis of known principles, facts, and subunits, it may have any degree of complexity without losing significance; the same is not true for analysis models. Significance can also be maintained by assembling a limited model to test a specific hypothesis of mechanism. PMID- 3036607 TI - Angiotensin-converting enzyme content of human spermatozoa and its release during capacitation. AB - In this study, we demonstrated, by using known detergents, the presence of angiotensin-converting enzyme (ACE) within human spermatozoa. We determined that maximal angiotensin-converting activity is expressed by sperm incubated in capacitating conditions, whereas this activity is negligible in saline-incubated spermatozoa. We further demonstrated that not acrosomes but cytoplasmic residues contain ACE. Because follicular fluid provides the necessary conditions for a maximal angiotensin-converting activity and for capacitations' metabolic activation, we hypothesize that ACE may play its physiologic role within the female reproductive tract. PMID- 3036608 TI - Mullerian aplasia associated with maternal deficiency of galactose-1-phosphate uridyl transferase. AB - The activity and electrophoretic pattern of galactose-1-phosphate uridyl transferase (transferase), a key enzyme in galactose metabolism, were analyzed in four patients with Mullerian aplasia (Rokitansky-Kuster-Hauser syndrome) and their mothers. Mothers of two of the patients had genetic variations of their transferase enzymes with activities below the normal range. Affected daughters from these two mothers also had genetic variations of the transferase enzyme. In one of the patients whose Mullerian aplasia had been diagnosed 15 years previously, premature ovarian failure developed. These case reports suggest a possible association between errors of galactose metabolism, Mullerian aplasia, and premature menopause--an association that is supported by a rodent model in which female offspring of mothers fed a high-galactose diet were born with reduced oocyte numbers and delayed vaginal opening. PMID- 3036609 TI - Cytomegalic virus infection in renal allograft recipients. Indicators for intervention in the SICU. PMID- 3036610 TI - Haematological and pathomorphological studies on chickens inoculated with transformation-defective sarcoma virus. AB - A transformation-defective mutant of Rous sarcoma virus (td PR-RSV-C) inoculated into chicken embryos caused a number of pathological changes in chickens after hatching. Disturbances in haematopoiesis were manifested by anaemia of different degree and by the presence of immature blood elements in the peripheral blood. The haematological picture of erythroblastosis was confirmed histologically. In addition to erythroblastosis, lymphoid leukosis and a proliferative lymphosarcoma like tumour were demonstrated histologically. PMID- 3036611 TI - Nonspecific cytotoxic cells in fish (Ictalurus punctatus). VI. Flow cytometric analysis. AB - Nonspecific cytotoxic cells (NCC) from channel catfish (Ictalurus punctatus) were enriched using flow cytometry (EPICS V, 753). Parameters of forward angle and 90 degrees light scatter were utilized to sort cytolytically functional populations of NCC. Fullbright (GR. II) 9.75 microns diameter fluorescent polystyrene microspheres were used for optical alignment. Cells obtained from the anterior kidney, spleen and peripheral blood were analyzed by one and/or two parameters of forward angle light scatter (FALS) and log integral 90 degrees light scatter (L90 degrees LS). Populations identified from FALS or L90 degrees LS histograms were sorted and cells were tested for cytotoxicity against NC-37 target cells. Anterior kidney contained at least four different sized populations of NCC. Spleen cells were sorted by FALS and four different populations of NCC detected. Peripheral blood contained little NCC activity. Enrichment of NCC activity from anterior kidney tissue was compared using Percoll density gradient separation and flow cytometry. Three to four fold higher cytotoxic activity was obtained from sorted cells compared to the population obtained from 45.5% Percoll. Additional experiments to determine the enrichment of sorted NCC were conducted by analyzing the number of enriched NCC required to kill an individual NC-37 target cell. Approximately 1.5-2.0 NCC could lyse a chromium-51 labelled target cell. These studies demonstrated that NCC could be enriched by size and granularity discrimination using flow cytometry. Effector:target cell ratios of 10:1 and lower produced significant lysis when sorted cells were analyzed. PMID- 3036612 TI - Decreased exchange of adenine nucleotides in human placental mitochondria. AB - The purpose of this work was to study the exchange of adenine nucleotides in mitochondria isolated from human placenta tissue. The results indicate that ADP and ATP are translocated at a lower rate than those reported for rat liver mitochondria. It is proposed that the limited transport is due to the particular lipid composition of placental mitochondria membrane, which induces an arrest in membrane fluidity with the consequent restriction in adenine nucleotide translocase mobility. PMID- 3036613 TI - D-galactosamine induced changes in rat liver plasma membranes lipid composition and some enzyme activities. AB - The influence of D-galactosamine administration on rat liver plasma membranes lipid composition, fluidity and some enzyme activities was investigated. D Galactosamine was found to induce an increase of the total phospholipids, the cholesterol level and membrane rigidity. In liver plasma membranes of D galactosamine-treated rats the exogenous phospholipase A2 activity was enhanced about 2 fold, whereas the endogenous activity was slightly decreased. No alteration of the neutral sphingomyelinase activity was observed. PMID- 3036614 TI - Purification and characterization of two phosphorylase phosphatases from rabbit liver. AB - Two phosphorylase phosphatase activities (I and III) have been purified from rabbit liver, with respective molecular weights of 117,000 and 230,000. Phosphatase III contained three different subunits of molecular weights 35,000, 67,000 and 80,000. Phosphatase I although majoritary in the preparation, was not homogeneous. Both phosphatases were dissociated by 2-mercaptoethanol treatment, releasing a catalytic subunit with a molecular weight of about 35,000. Phosphatases I and III activities responded very differently to incubation with trypsin and to ethanol precipitation. Phosphatase III was much more sensitive to inactivation by several ions and ATP than phosphatase I. On the basis of the obtained data, phosphatase I can be classified as a type-1 phosphatase and phosphatase III as a type-1 phosphatase. PMID- 3036615 TI - Characterization of dolichol kinase from bovine thyroid microsomes. AB - Bovine thyroid microsomes are able to phosphorylate exogenous [1-3H]dolichol as well as endogenous dolichol. The properties and specificity of the dolichol kinase activity have been studied by following the phosphorylation of [1 3H]dolichol to [1-3H]DMP as well as the formation of [32P]DMP from endogenous dolichol and [gamma-32P]CTP. The dolichol kinase activity was not linear with respect to time and exhibited a neutral pH-optimum. Product formation was directly proportional to microsomal protein concentration up to 2.5 mg protein/incubation. The enzyme was found to depend on divalent cations for activity: Mg2+-ions being much more effective than Ca2+- and Mn2+-ions. In accordance, EDTA was strongly inhibitory. The enzyme exhibited specificity for CTP as phosphoryl donor and was found to be inhibited by the reaction product CDP. The apparent Km-value for exogenous dolichol amounted to 4 microM. Those for CTP were estimated to be 3.88 and 10.75 mM with exogenous [1-3H]dolichol depending on the source of CTP. With endogenous dolichol Km-values for CTP of 27.8 and 6.1 microM were calculated in respectively the absence and presence of 5 mM VO4(3-). Triton X-100 (0.15%) was necessary in the [1-3H]dolichol kinase assay (only 3% of enzymatic activity in the absence of detergent), while with [gamma 32P]CTP dolichol kinase detergent was only of minor influence (30% stimulation at 0.02% Triton X-100). The levels of the enzymatic activity could be doubled by the inclusion of 18-21 mM NaF [( 1-3H]dolichol kinase) as phosphatase inhibitor: VO4(3-) had practically no effect. In contrast with [gamma-32P]CTP dolichol kinase, the enzymatic activity could be enhanced 4-fold by addition of 5 mM VO4(3 ) while F- resulted into no appreciable effect.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3036616 TI - Identification and characterization of a specific dolichylmonophosphate phosphatase activity in bovine thyroid microsomes. AB - Bovine thyroid membranes are able to dephosphorylate exogenous [1-3H]DMP as well as endogenous prelabeled [32P]DMP. The kinetics, properties and specificity of the dolichylmonophosphatase activity have been studied by monitoring respectively the release of [1-3H]dolichol from [1-3H]DMP and the residual amount of [32P]DMP. The DMP-phosphatase activity is not linear with respect to time and exhibits a neutral pH optimum. There is only linearity in a narrow range of protein concentration when 0.25% Triton X-100 is included in the incubation mixture. Studying the enzymatic activity in function of protein concentration, the detergent requirement shows to be very critical. Triton X-100 is necessary for enzymatic activity with [1-3H]DMP (only 10% of enzymatic activity in the absence of detergent) although the detergent inhibits the hydrolysis of endogenous prelabeled [32P]DMP. Divalent cations are not essential for enzymatic activity, Ca2+-ions being even inhibitory. In accordance, EDTA (EGTA) is slightly stimulatory. The DMP-Pase activity is not influenced by the ionic strength of the incubation system and sulphydryl groups are not involved. NaF, VOS and VO4(3-) are strongly inhibitory. The inhibition by dolichol and PO3-4 can be explained as the result of product inhibition. An apparent Km-value of 2.5 X 10(-5) M is computed for [1-3H]DMP. Bacitracin inhibits DMP-phosphatase in contrast with other reports. Propylthiouracyl, cAMP, TSH and several other bio-effectors are without effect on the in vitro system. The specificity of the DMP-Pase activity is discussed, showing that the phosphatase is distinctly different from other phosphatases especially phosphatidic acid phosphohydrolase. PMID- 3036617 TI - Stimulation by glucagon and adenosine-3',5'-cyclic monophosphate of protein degradation in Reuber H35 hepatoma monolayers. AB - Protein degradation in Reuber H35 hepatoma monolayers was measured as release of radioactive trichloroacetic acid-soluble material from intracellular protein labelled with [3H]leucine for 16 hr followed by 3-hr chase period. Proteolysis in this system was stimulated by physiological concentration of glucagon reaching a maximum at 10(-7) M with an increase of 30%. Dibutyryl cyclic AMP also had a stimulatory effect. When both glucagon and dibutyryl cyclic AMP were present at optimal concentrations, their effects were not additive suggesting that glucagon may act via the formation of cyclic AMP. In the presence of protein synthesis inhibitor, cycloheximide or puromycin, proteolysis remained responsive to glucagon. Glucagon counteracted the inhibitory effect of insulin on proteolysis. PMID- 3036618 TI - Separation and characteristics of inside-out and right side-out vesicles from a rat cardiac sarcolemma preparation. AB - Purified cardiac sarcolemma (SL) vesicles are highly suitable to study various Ca2+-transport systems present in the SL. We describe in this paper the separation of the Inside-Out (IO) and Right side-Out (RO) oriented vesicle subpopulations from a purified rat heart SL preparation. The isolated subfractions were characterized with respect to the number of beta-adrenergic binding sites and the Ca2+-uptake and (Ca2+-Mg2+)-ATPase activities. It was found that the Ca2+-uptake and the (Ca2+-Mg2+)-ATPase activities reside in the IO fraction and are virtually absent in the RO fraction, confirming that the active Ca2+-uptake represents the outward directed sarcolemmal Ca2+-flux. PMID- 3036619 TI - Stimulation of mitochondrial functions by glucagon treatment, starvation and by treatment of isolated mitochondria with glycogen-bound enzymes. AB - Liver mitochondria isolated from rats starved overnight, or fed rats injected with glucagon, exhibited a similar increase of the respiration rate with succinate (by 30-40%) and glutamate plus malate (by 20-30%), as compared to mitochondria from control fed animals. The content of mitochondrial adenine nucleotides was elevated by 30-45% by glucagon treatment or starvation. Mitochondrial respiration and citrulline synthesis were stimulated by 30-40% when mitochondria isolated from fed rats were briefly preincubated with the extract from liver glycogen granules, ATP and MgCl2. This effect was abolished by heating the extract at 100 degrees C. PMID- 3036620 TI - Comparative study of the developmental patterns of vasopressin, glucagon, angiotensin II, and alpha 1-adrenergic receptors in the liver of developing and adult hypothyroid rats. AB - The effects of propylthiouracil (PTU) treatment on vasopressin, angiotensin II, glucagon and alpha 1-adrenergic receptors in both developing and adult rats were studied in liver membrane preparations by measuring the binding of the following ligands: [3H][8-lysine]vasopressin, [3H]Sar-angiotensin II, [125I]glucagon and [3H]prazosin, and in the case of glucagon, by measuring adenylate cyclase activation. Whatever the ligand used, in young as well as in adult animals, PTU treatment led to a similar reduction (about 50%) in the maximal number of binding sites (Bmax), without significant changes in the apparent dissociation constant (KD) of labeled hormone for its specific receptor. In normal adult animals, thyroxine treatment, i.e. hyperthyroidism, had an opposite effect on the Bmax (25 50% increase), without changes in the KD. In developing PTU-treated rats, the abnormalities completely disappeared after therapy with increasing physiological doses of thyroxine; consequently they were directly related to thyroid deficiency and not to toxic effects of PTU. Moreover, the abnormalities resulting from induced hypothyroidism were reversible. In developing and adult hypothyroid rats, neither basal, NaF-, nor Gpp(NH)p-stimulated adenylate cyclase activities were significantly affected. Glucagon-sensitive adenylate cyclase activity seemed to be slightly increased (by about 15%), without changes in the apparent activation constant (Kact). These results are considered in parallel with findings on plasmatic glucagon and vasopressin levels, compared with similar previous reports related to renal vasopressin receptors, and discussed with respect to unpublished observations concerning hepatic responsiveness to glycogenolytic hormones in young and adult rats with induced hypothyroidism. PMID- 3036621 TI - Thyroid hormone receptors in a human hepatoma cell line: multiple receptor forms on isoelectric focusing. AB - The nuclear thyroid hormone receptors isolated from cultured human hepatoma cells (Hep G2) were characterized and compared with those from cultured human fibroblasts and rat liver. The Hep G2 nuclear thyroid hormone receptors had an affinity constant (Ka) of 2.1 X 10(10) M-1 and maximal binding capacity (MBC) of 21.0 fmol/100 micrograms DNA for T3 in assays performed on isolated nuclei. 16% of nuclear receptors were released into the media during incubation and had the same Ka. Salt-extracted receptors had a Ka of 1.8 X 10(10) M-1 and MBC of 0.1 pmol/mg protein for T3. Density gradient sedimentation and gel filtration chromatography revealed a sedimentation coefficient of 3.4 S and Stokes radius of 34 A. From these values, a molecular weight of 49,000 and total frictional ratio (f/f0) of 1.4 were calculated, suggesting an asymmetrical shape of the receptor molecule. Heat inactivation occurred with t1/2 of 28.1, 18.0, and 7.9 min at 38, 43, 45 degrees C, respectively. Isoelectric focusing (IEF) of Hep G2 nuclear receptors demonstrated T3 binding proteins at pH 5.3-5.5, 5.7, and 5.9. Evidence that these are nuclear thyroid hormone receptors includes the following: Triiodothyroacetic acid was the most potent competitor of [125I]T3 binding to these proteins followed by L-T3, and L-T4. Cytosolic protein, human serum, and fetal calf serum failed to show the same T3 binding proteins. Ka of these proteins measured by T3 displacement was 1.1-3.2 X 10(9) M-1. Human fibroblast nuclear extract showed similar T3 binding pattern in IEF, except for a slight difference in pI of an acidic band.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3036622 TI - Proliferative response to cyclic AMP elevation of thyroid epithelium in suspension culture. AB - The role of cyclic AMP (cAMP) as an intracellular growth signal in thyroid epithelial (follicular) cells has been studied using primary suspension cultures of rat thyroid follicles closely resembling the in vivo state. TSH at 0.1 mU/ml in the presence of insulin (0.08 microgram/ml) induced a 9.6-fold increase in [3H]thymidine incorporation with a peak after 48-72 h. The response to the cAMP elevating agent cholera toxin (10 ng/ml) was identical both in timing, magnitude and dependence on insulin. A lower amplitude response occurred with forskolin. The data further support the conclusion that elevation of intracellular cAMP concentration is a major, if not the only, signal required for the proliferative response of the follicular cell to its physiological mitogen, TSH. PMID- 3036623 TI - Inhibitory and stimulatory effects of dexamethasone and 1,25-dihydroxyvitamin D3 on chick intestinal calbindin-D28K and its mRNA. AB - The present study examined the interactions of the synthetic glucocorticoid, dexamethasone, with the regulation of chick intestinal calbindin-D28K (a 28,000 Da vitamin D-dependent calcium binding protein, CaBP) and its mRNA by 1,25 dihydroxyvitamin D3 (1,25(OH)2D3). Dexamethasone (0-500 nmol) had a neutral impact on calbindin levels in the rachitic chick intestine when measured 12 h later. However, dexamethasone appeared to exert a significant, though modest, stimulatory influence upon calbindin-mRNA accumulation in the vitamin D-deficient (-D) intestine when measured 12 h after administration. 1,25(OH)2D3 also stimulated calbindin-mRNA accumulation in the -D chick intestine; half-maximal (ED50) doses were 1.1 nmol (7.6-fold) and 12.6 nmol (4.3-fold stimulation) for 1,25(OH)2D3 and dexamethasone respectively. In contrast, when both 1,25(OH)2D3 and dexamethasone were administered simultaneously, the stimulatory effect of 1,25(OH)2D3 (and that of the glucocorticoid) was lost in terms of calbindin and calbindin-mRNA accumulation. Dexamethasone treatment of vitamin D-replete (+D) chicks resulted in a depression of calbindin-mRNA accumulation; levels were depressed to baseline with 250 nmol/bird. Dexamethasone (1.25 mumol per day for 3 days) also induced an apparent 'down-regulation' of the 1,25(OH)2D3 receptor population in the -D chick intestine but failed to influence the binding of 1,25(OH)2D3 to its receptor in vitro. Taken collectively, these data indicate that glucocorticoids are able to influence the receptor-mediated action of 1,25(OH)2D3, possibly at the level of calbindin-D28K gene expression. PMID- 3036624 TI - Action mechanism of inhibin in vitro--cycloheximide mimics inhibin actions on pituitary cells. AB - The effects of inhibin on gonadotropin secretion from cultured rat anterior pituitary cells were examined by using purified porcine follicular fluid (pFF) 32 kDa inhibin. pFF 32 kDa inhibin suppressed the basal FSH secretion as well as cell content of FSH with identical ED50 values (ED50 = 1.0 ng/ml) in a dose dependent manner, but did not alter either basal secretion or cell content of LH. On the other hand, pretreatment of the pituitary cells with pFF 32 kDa inhibin during the first 3 days resulted in suppression of subsequent LH-RH-stimulated release of both FSH and LH in a dose-dependent manner, indicating that the suppression of LH-RH-stimulated release of LH is one of the intrinsic inhibin actions on pituitary cells. The marked difference between ED50 values for FSH (ED50 = 1.1 ng/ml) and LH (ED50 = 2.5 ng/ml) in the suppression of LH-RH stimulated release of gonadotropins, together with the fact that the total amount of LH (cell content plus released) after LH-RH stimulation remained unchanged following inhibin treatment suggests that the suppression of LH-RH-stimulated release of LH by inhibin is quite different from that of FSH regarding the action mechanism. Similarly, cycloheximide, an inhibitor of protein biosynthesis, suppressed both basal secretion and cell content of FSH with almost the same ED50 values (ED50 = 22.5 ng/ml) but did not alter either basal secretion or cell content of LH. Cycloheximide also suppressed LH-RH-stimulated release of both FSH and LH, and the ED50 values were different from each other (ED50 = 25.0 ng/ml for FSH and 60.0 ng/ml for LH suppression, respectively). Our finding that cycloheximide completely mimicked the action of inhibin on gonadotropin secretion strongly suggests that LH is quite insensitive to biosynthetic inhibition, and that preferential effects of inhibin or cycloheximide on FSH in appearance may reflect the difference between LH and FSH in susceptibility to biosynthetic inhibition. PMID- 3036625 TI - The mechanism of angiotensin-induced desensitization of adrenal glomerulosa cells. AB - Superfusion of isolated rat adrenal glomerulosa cells for 6 h with a medium containing 2.5 nM angiotensin II (AII) reduces the aldosterone response to AII, corticotropin and potassium. Here we report that under such conditions there is a decrease in the capacity of the cells to form inositol phosphates in response to a subsequent stimulation with AII. The capacity to convert corticosterone to aldosterone is also reduced by a prior exposure to AII. Superfusion with a high potassium medium has no such an effect. Reduced phosphoinositide response may be responsible for the decreased aldosterone stimulation by AII, the inhibition of the late stage of aldosterone biosynthesis may account for the heterologous character of desensitization. PMID- 3036626 TI - Regulation of calcium fluxes in the thyroid. AB - Calcium (Ca2+) exchanges were studied in dog thyroid slices incubated in vitro. With 45Ca2+-prelabeled slices, carbamylcholine 10(-7)-10(-5) M (Cchol) induced an important transitory spike efflux, inhibited by procaine and atropine while the stimulated efflux obtained with high concentrations of TSH (10 mU/ml) was progressive and sustained over time. The effects observed with both agents did not require extracellular Ca2+ and were insensitive to verapamil 10(-6)-10(-4) M. Neither dibutyryl (Bu2)-cAMP, nor any agent raising intracellular cAMP (prostaglandin E2, choleratoxin, inhibitors of phosphodiesterases with low concentrations of TSH) were able to reproduce the action of TSH 10 mU/ml, forskolin 10(-5) M being the only exception. Replacement of sodium by choline (+ atropine) in the incubation medium decreased the basal efflux and inhibited the TSH effect. Ouabain 10(-3) M also abolished the TSH-induced Ca2+ efflux, while having no influence on carbamylcholine action. TSH 10 mU/ml and 1 mU/ml, Bu2-cAMP 10(-3) M, choleratoxin and prostaglandin E2 with inhibitors of phosphodiesterase decreased the total 45Ca2+ uptake of the slices, while no effect of Cchol could be detected on this parameter. The results obtained suggest that (1) Cchol and TSH stimulate 45Ca2+ efflux from dog thyroid slices with different kinetics, by mobilization of intracellular Ca2+ stores; (2) this effect of TSH is not mediated by cAMP; (3) independently TSH at low concentrations (1 mU/ml), through cAMP, decreased 45Ca2+ uptake; this suggests that increased 45Ca2+ efflux and decreased uptake result from different mechanisms, as has been described for iodide exchange in FRTL-5 cells. PMID- 3036627 TI - Androgen receptor-binding regions of an androgen-responsive gene. AB - The interaction of 5 alpha-dihydrotestosterone-receptor complexes with purified DNA fragments representing upstream, coding and intervening sequences of the prostate binding protein C3(1) gene was investigated using a DNA-cellulose competition binding assay. The partially purified androgen-receptor complexes which were used in the assay had proven DNA-binding capabilities. Two fragments were identified with relatively high affinity for androgen-receptor complexes. A 300 bp fragment extending from -220 to +80 and a 500 bp fragment derived entirely from the first intron consistently competed most effectively in the system. The presence of a high affinity site or sites in or near the promoter region of the gene is consistent with current models of transcriptional activation of hormone responsive genes by steroid receptors. High affinity sites for steroid receptors within introns may indicate a role for receptors in regulation of transcription at other stages, or in post-transcriptional modification. PMID- 3036628 TI - Phorbol ester stimulates human granulosa-luteal cell cyclic adenosine 3', 5' monophosphate and progesterone production. AB - Human granulosa-luteal cell production of cyclic adenosine 3', 5'-monophosphate (cAMP) and progesterone (P) were studied in response to 12-O-tetradecanoyl phorbol-13-acetate (TPA). TPA specifically increased cAMP synthesis in a dose dependent manner. A 7-fold increase occurred at a TPA concentration of 1 ng/ml. Time-course studies indicated that the increase in accumulation of cAMP into culture media became detectable at 4 h and continued up to 72 h. TPA also enhanced P synthesis, but the increase was statistically significant only at 72 h. Indomethacin prevented TPA-stimulated cAMP and P production. The results suggest that TPA stimulates granulosa-luteal cell cAMP and P production, and that the action of TPA is mediated by the increase in prostaglandin synthesis. PMID- 3036629 TI - Development of physiological responsiveness to glucagon during embryogenesis of avian heart. AB - Glucagon increases contractility of the heart muscle by stimulation of adenylate cyclase activity and elevation of cAMP. We have investigated the specific time of onset of glucagon sensitivity of heart muscle during development of the chick embryo. Using both isolated heart preparation and cultured cardiac cells, we have found that the contractile response to glucagon cannot be detected prior to Day 4 of development. Binding studies, carried out with heart cells prepared from 3-, 5 , 7-, and 11-day chick embryos, showed a significant increase in the number of glucagon binding sites between Days 3 and 5. Scatchard analysis showed that for Day 5 cells maximum binding capacity was 0.56 pmole/mg of protein with Kd of 16.0 nM, while for Day 3, maximal binding was only 0.16 pmole/mg with Kd of 15.1 nM. Therefore in this 2-day interval there was a marked increase in the receptor number, without changes in the receptor affinity. Since hormonal stimulation of adenylate cyclase depends on the presence of the regulatory component (Ns), we have used cholera toxin-induced chronotropic effect as an assay for functional Ns. No response to cholera toxin could be detected prior to Day 4 of chick heart development. Therefore, emergence of the cholera toxin sensitivity correlates well with the onset of responsiveness to glucagon. We conclude that as the heart develops it acquires a physiological responsiveness to glucagon. The acquisition of the hormonal sensitivity correlates with the increase in the receptor number and the functional levels of regulatory component. PMID- 3036630 TI - Different molecular mechanisms for cAMP regulation of gene expression during Dictyostelium development. AB - Alterations in cAMP concentrations have been implicated in developmentally regulated gene expression in Dictyostelium. Using a variety of culture conditions to control the metabolism of cAMP during cytodifferentiation, I have examined the role of the cyclic nucleotide in development. Conditions which allow intracellular synthesis of cAMP promote the normal developmental repression of gene M4-1 by a mechanism which is completely independent of the formation of multicellular aggregates. If, however, cells are inhibited in their ability to activate adenylate cyclase and, thus, intracellular cAMP signaling, they prove unable to repress M4-1, even in the presence of exogenous cAMP. In contrast, expression of genes which exhibit maximal activity after aggregate formation depends upon accumulation of extracellular cAMP. Inhibition of intracellular cAMP signaling does not prevent the expression of these genes if cultures are simultaneously exposed to high levels of exogenously added extracellular cAMP. These results indicate that there are at least two independent mechanisms involved in the developmental regulation of gene expression by cAMP in Dictyostelium. I discuss plausible molecular mechanisms through which cAMP might alter gene expression. PMID- 3036631 TI - The argentia mutation delays normal development of photosynthetic cell-types in Zea mays. AB - Photosynthesis in C4-type grasses such as maize involves the interaction of two cell types (bundle sheath (BS) and mesophyll (M)) which both contain cell specific photosynthetic enzymes. Malate dehydrogenase, phosphoenolpyruvate carboxylase and pyruvate phosphate dikinase are located in the M cells and malic enzyme and ribulose bisphosphate carboxylase are found in the BS cells. We have studied photosynthetic development in leaves of the temperature-sensitive greening mutant argentia (ar). We have determined that with the exception of malate dehydrogenase, levels of C4 enzymes are lower in ar leaves than in normal leaves. Malate dehydrogenase accumulates identically in both. Using in situ immunolocalization techniques with normal and ar leaves, we have observed a developmental pattern of C4 protein accumulation. In normal leaves protein was detected first in cells surrounding the median vein, then in cells surrounding other major veins, and finally in cells surrounding minor veins. In ar leaves, C4 enzymes accumulate in the correct cell type and in this same order but their appearance is delayed. Furthermore, BS cell development is delayed with respect to M cell development. The observed pattern of photosynthetic development reflects an earlier manifested pattern of vascular development yet the timing of vascular differentiation in ar mutants appears to be normal. PMID- 3036632 TI - Fetomodulin: marker surface protein of fetal development which is modulatable by cyclic AMP. AB - A novel cell surface marker of fetal development was identified in both in vivo and in vitro systems of the mouse using monoclonal antibodies against a glycoprotein of an apparent size of 133,000 Da. Two independent clones of hybridomas were isolated by fusing murine myeloma cells, NS-1, with spleen cells of a rat which was immunized with murine 3T3 fibroblast. The analysis of molecular size and tryptic peptides of the immunoprecipitate indicated that fibroblast and putative parietal endoderm cells, which were derived by induced differentiation of F9 embryonal carcinoma cells with retinoic acid and cyclic AMP, expressed apparently the same protein. Undifferentiated F9 cells and F9 cells which were treated with retinoic acid or cyclic AMP alone had little or no immunoprecipitable proteins. Analogously, parietal endoderm of in vivo embryos tested positive for this protein but visceral endoderm and embryonic ectoderm did not. The amount of this surface protein was increased in fibroblast and differentiated F9 cells by elevation of intracellular cyclic AMP concentrations. These results are consonant with a hypothesis that this surface protein plays a role in fetal development via a quantitative modulation by cyclic AMP. PMID- 3036633 TI - Synapse formation by sensory neurons after cross-species transplantation in crickets: the role of positional information. AB - The role of positional information in synapse formation was studied in the cricket cercal sensory system by transplanting epidermis from one species of cricket to another. Strips of cercal epidermis containing identified sensory neurons were transplanted from a black donor species to a tan host species; the color difference was used to distinguish between donor and host tissue in adults. Transplanted sensory neurons regenerated axons into the host terminal abdominal ganglion where they formed functional chimeric synapses. These methods were used to test the role of positional information in central synapse formation. Newly generated sensory neurons, formed by the donor tissue at the border between graft and host, were examined to test the idea that their position would determine their structure, function, and projection pattern. These "intercalated" sensory neurons support the positional information hypothesis. First, they had directional sensitivities which were appropriate to their location on the cercus; receptors of this directionality would never be made by the donor tissue if left in its original position. Second, these sensory neurons projected to regions of the CNS known to be appropriate for their directionality. Finally, simultaneous recordings from these ectopic sensory neurons and host interneurons demonstrated the expected synaptic connection, based on the overlap of pre- and postsynaptic cells. Thus three aspects of receptor function, directionality, afferent projection, and choice of synaptic partners, appeared to be controlled by positional information. PMID- 3036634 TI - Differential effect of steady-state hyperinsulinaemia and hyperglycaemia on hepatic glycogenolysis and glycolysis in rats. AB - The action of glucose and of insulin on hepatic glucose production and metabolism has been studied in fed anaesthetized rats during hyperinsulinaemic clamp combined with various steady state levels of glycaemia (6.8 +/- 0.1, 9.3 +/- 0.1, 11.8 +/- 0.1 mmol/l). Hepatic glucose production was measured using constant infusion of D-[6-3H] glucose. At the end of each clamp the liver was freeze clamped, and enzyme activities and metabolites were measured. Hepatic glucose production was totally suppressed in all the groups receiving insulin. In the group with steady-state normoglycaemia, the suppression of hepatic glucose production was accompanied by a decrease in the levels of glucose-6-phosphate, an increase in those of fructose 2,6-bisphosphate and glycolytic intermediates, but without change in glycogen level or glycogen synthase and phosphorylase. In contrast, in the groups with steady-state hyperglycaemia, phosphorylase a was inactivated, and glycogen synthase activated. Under these conditions, glucose-6 phosphate levels were also decreased and those of fructose 2,6-bisphosphate and glycolytic intermediates were higher than in the group with steady-state normoglycaemia. A slight drop in the level of cAMP was also observed which may contribute, with hyperglycaemia, to the inactivation of phosphorylase. Incorporation of tritiated water into liver glycogen paralleled the activation of glycogen synthase and the accumulation of glycogen. The data indicate that, at normoglycaemia, insulin may suppress hepatic glucose production by channeling glucose-6-phosphate into the glycolytic pathway.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3036635 TI - Distinct developmental regulation and properties of the responsiveness of different genes to cyclic AMP in Dictyostelium discoideum. AB - Cyclic AMP (cAMP) is known to be an important mediator of gene expression in eukaryotic cells. At present, little is known about the developmental events which render specific genes responsive to cAMP in distinct cell types, or about the biochemical mechanisms by which cAMP exerts these regulatory effects. By examining the effects of cAMP treatment on specific mRNA levels in Dictyostelium discoideum cells with different 'developmental histories', we defined the developmental states in which specific genes display responsiveness to cAMP. We focused on two specific rapid responses: the ability of cAMP to inhibit the expression of an 'early' developmentally regulated mRNA (discoidin-I) and to stimulate the expression of a 'late', prespore-specific mRNA (PL3). Using this approach, we showed that, for both mRNAs, the ability to respond rapidly to cAMP is absent from vegetative cells grown on bacteria, and is acquired during development on filters. Furthermore, we identified several developmental states in which the discoidin-I response to cAMP is present, but in which the PL3 response is not. In experiments designed to examine the effects of cAMP analogues on the levels of these two mRNAs, we demonstrated that the analogue specificities of the discoidin-I and PL3 responses are different, and that the specificity for the PL3 response depends on the developmental state. The developmental kinetics and analogue specificity of the PL3 response suggest a two-step mode of action of cAMP in activating the expression of this gene. We discuss possible implications of these findings for the mechanisms of action of exogenous cAMP as well as for the role of cAMP in controlling the changes in gene expression that accompany normal development. PMID- 3036636 TI - Fistula from the superior mesenteric artery to duodenum: a rare cause of death from pancreatic carcinoma. AB - A fistula from the superior mesenteric artery to the duodenum is an extremely rare complication of pancreatic carcinoma. It presents with overwhelming upper gastrointestinal hemorrhage and can lead to death from blood loss. PMID- 3036637 TI - Cholangiocarcinoma arising in a choledochal cyst: diagnostic value of computed tomography and ultrasonography. AB - A case of cholangiocarcinoma arising in a choledochal cyst (type II or biliary cyst in Alonso-Ley classification) in a 53-year-old woman is presented. Ultrasonography and computed tomography provided the correct preoperative diagnosis. The value of these modalities in presurgical planning is illustrated. PMID- 3036639 TI - Ricinus communis agglutinin II-reactive glycoproteins from the ascites of patients with hepatocellular carcinoma and their use in enzyme-linked immunosorbent assay. AB - Ricinus communis agglutinin II-reactive glycoproteins from the ascites of patients with hepatocellular carcinoma were prepared using lectin affinity chromatography. Normal serum- and cirrhotic ascites-components were removed by columns with immobilized antibodies against them. Ricinus communis agglutinin II reactive glycoproteins thus obtained were supposed to be hepatocellular carcinoma associated and less than 0.1% of the protein in the starting material. Polyacrylamide gel electrophoresis of these glycoproteins revealed more than 10 major polypeptides with molecular weights ranging from 20K to 200K daltons. The rabbit antiserum raised against them reacted with at least three components of 45, 52 and 55K daltons. The serum level of this antibody-reactive glycoproteins was assessed by an enzyme-linked immunosorbent assay. It was elevated in 91% of cases of hepatocellular carcinoma, 70% of cases of other gastrointestinal carcinoma, 88% of cases of liver cirrhosis, 55% of cases of chronic hepatitis, and 25% of cases of acute hepatitis. The mean value of hepatocellular carcinoma was significantly greater than those of other groups. These results suggest that some of Ricinus communis agglutinin II-reactive glycoproteins in hepatocellular carcinoma patients may be cancer-associated glycoproteins and that their serum levels are increased in hepatocellular carcinoma patients. PMID- 3036638 TI - In vitro interferon producing activity of peripheral mononuclear cells in patients with chronic liver disease. AB - Interferon-gamma (IFN-gamma) was induced from a human peripheral mononuclear fraction by incubation with a streptococcal preparation stabilized with penicillin G (OK432). This IFN-gamma-producing activity was significantly reduced in patients with chronic hepatitis and hepatocellular carcinoma. In patients with liver cirrhosis it was also reduced but not significantly. Serum hepatitis B virus DNA and skin tests for the purified protein derivative of tuberculin, phytohemagglutinin-P and a polysaccharide fraction prepared from streptococcus pyogenes Su strain were determined to have no significant relation to this IFN gamma-producing activity. Although the addition of interleukin 2 (IL-2) to the culture medium enhanced the IFN-gamma-producing activity, there was no difference in this enhancement between normal control and chronic hepatitis. Therefore reduction of the IFN-gamma-producing activity observed in chronic hepatitis seems to be caused by a decreased number of IFN-gamma-producing activity cells or hypofunction of these cells or both. Since HBeAg became negative in patients whose IFN-gamma-producing activity was increased by the administration of the immunopotentiator OK432 or IFN-beta, the IFN-producing system in the patients with B type hepatitis may contribute to the elimination of HBV. Adenine arabinoside suppressed IFN-gamma-producing activity both in vivo and in vitro. PMID- 3036641 TI - [Pregnancy following cytostatic treatment of trophoblast tumor]. AB - Observation of 18 pregnancies in 13 women who had undergone treatment for trophoblastic neoplasm with cytostatic drugs confirmed reports according to which the risk of miscarriage, premature birth, stillbirth, and severe congenital anomalies is not increased. Seven low-risk patients who had been treated with methotrexate gave birth to 10 children who were mature except for one pair of premature twins. Only one child had an anomaly, i.e. celiac disease, but this could not be linked to the prior treatment. Three high-risk patients who had been treated with methotrexate or methotrexate with actinomycin-D gave birth to four children, all of whom were mature and healthy. Intensive prenatal care, and in particular diagnosis at the beginning of pregnancy, are of great importance. At least one year must elapse between termination of chemotherapy and the beginning of a pregnancy. Effective contraception must be assured during this time. PMID- 3036640 TI - [Immunohistochemical diagnosis of occult breast cancer]. AB - The techniques associated with immunohistochemistry are of major interest in modern tumour diagnosis. Their simple application provides a wider scope for diagnosis by increasing the possibilities of characterising tumour cells. In particular, the combination of various tumour markers has led to a better understanding of tumour histogenesis. If the primary tumour is unknown and only metastases are available for diagnosis, further important information can be gained allowing a correlation with the origin of the metastases. In three own cases an occult carcinoma of the female breast could be diagnosed pre-operatively via immunohistochemistry techniques. Since no marker is available that reacts specifically with cells of metastasizing mammary cancer, lymph node metastases have been examined with several different markers. For that purpose, the epithelial membrane antigen (EMA), Thomsen-Friedenreich-antigen (T-Ag) and alpha lactalbumin (ALA) have been used to point out several pathways of cell metabolism. It could be demonstrated that the prospective use of the three markers is of the highest value to make diagnosis easier and safer than by interpreting the morphological appearance. The diagnostic value of tumour markers is an important step forward, especially in the interpretation of tumour cell embolism, macrometastases and micrometastases in regional lymph nodes. PMID- 3036642 TI - Modification of primary amino groups in rat heart sarcolemma by 2,4,6 trinitrobenzene sulfonic acid in respect to the activities of (Na+ + K+)-ATPase, Na+-ATPase and pNPPase. Function of the potassium binding sites. PMID- 3036643 TI - Polymorphism and linkage of the alpha A-crystallin gene in t-haplotypes of the mouse. AB - Restriction fragment polymorphisms were used to order the alpha A-crystallin locus (Crya-1) relative to other genes in mouse t-chromatin and to investigate the relatedness of alpha-A-crystallin sequences among different t-haplotypes. Analysis of DNA from t-recombinant mice mapped Crya-1 to the K end of the H-2 complex and within the distal inverted region characteristic of t-haplotypes. Hybridization with Crya-1 cDNA revealed three distinct phenotypic groups among the 17 different t-haplotypes studied. A majority (9 of 17) of the t-haplotypes were classified into a novel group (Crya-1t) characterized by restriction fragments apparently unique to t-chromosomes and therefore thought to contain alpha A-crystallin sequences descended from the original t-chromosome. A second group of t-haplotypes had restriction fragment patterns indistinguishable from those observed among many common inbred strains of mice of the Crya-1a type, and a third restriction fragment pattern, observed only in the tw121 haplotype, was indistinguishable from the fragment pattern for C3H/DiSn (Crya-1b) and several other inbred strains of mice. Thus, with respect to sequences around the Crya-1 locus, different t-haplotypes show restriction fragment polymorphisms, some of which are comparable to those found in wild-type chromosomes and provide further evidence for genetic heterogeneity in DNA from the distal region of t-haplotypes. PMID- 3036644 TI - The constitutive, glucose-repression-insensitive mutation of the yeast MAL4 locus is an alteration of the MAL43 gene. AB - Mutations resulting in constitutive production of maltase have been identified at each of the five MAL loci of Saccharomyces yeasts. Here we examine a dominant constitutive, glucose-repression-insensitive allele of the MAL4 locus (MAL4-C). Our results demonstrate that MAL4-C is an alteration in the MAL43 gene, which encodes the positive regulator of the MAL structural genes, and that its product is trans-acting. The MAL43 gene from the MAL4-C strain was cloned and integrated into a series of nonfermenting strains lacking a functional regulatory gene but carrying copies of the maltose permease and maltase structural genes. Expression of the maltase structural gene was both constitutive and insensitive to glucose repression in these transformants. The MAL4-C allele also results in constitutive expression of the unlinked MAL12 gene (encoding maltase) in this strain. In addition, the cloned MAL43 gene was shown to be dominant to the wild-type MAL63 gene. We also show that most of the glucose repression insensitivity of strains carrying the MAL4-C allele results from alteration of MAL43. PMID- 3036645 TI - Mutations affecting expression of the rosy locus in Drosophila melanogaster. AB - The rosy locus in Drosophila melanogaster codes for the enzyme xanthine dehydrogenase (XDH). Previous studies defined a "control element" near the 5' end of the gene, where variant sites affected the amount of rosy mRNA and protein produced. We have determined the DNA sequence of this region from both genomic and cDNA clones, and from the ry+10 underproducer strain. This variant strain had many sequence differences, so that the site of the regulatory change could not be fixed. A mutagenesis was also undertaken to isolate new regulatory mutations. We induced 376 new mutations with 1-ethyl-1-nitrosourea (ENU) and screened them to isolate those that reduced the amount of XDH protein produced, but did not change the properties of the enzyme. Genetic mapping was used to find mutations located near the 5' end of the gene. DNA from each of seven mutants was cloned and sequenced through the 5' region. Mutant base changes were identified in all seven; they appear to affect splicing and translation of the rosy mRNA. In a related study (T. P. Keith et al. 1987), the genomic and cDNA sequences are extended through the 3' end of the gene; the combined sequences define the processing pattern of the rosy transcript and predict the amino acid sequence of XDH. PMID- 3036646 TI - Sequence of the structural gene for xanthine dehydrogenase (rosy locus) in Drosophila melanogaster. AB - We determined the nucleotide sequence of a 4.6-kb EcoRI fragment containing 70% of the rosy locus. In combination with information on the 5' sequence, the gene has been sequenced in entirety. rosy cDNAs have been isolated and intron/exon boundaries have been determined. We find an open reading frame which spans four exons and would encode a protein of 1335 amino acids. The molecular weight of the encoded protein (xanthine dehydrogenase), based on the amino acid translation, is 146,898 daltons which agrees well with earlier biophysical estimates. Characteristics of the protein are discussed. PMID- 3036648 TI - IS50L as a non-self transposable vector used to integrate the Bacillus thuringiensis delta-endotoxin gene into the chromosome of root-colonizing pseudomonads. AB - Insertion sequence IS50L of transposon Tn5 was used as a non-self transposable vector to integrate the delta-endotoxin gene (tox) from Bacillus thuringiensis subsp. kurstaki HD-1 into the chromosome of two corn-root colonizing strains of Pseudomonas fluorescens (112-12 and Ps3732-3-7). A DNA fragment containing the KmR gene from Tn5 and tox was inserted into an IS50L element (IS50L-tox) contained on a suicide plasmid. Transposition of IS50L-tox into the chromosome of P. fluorescens 112-12 and Ps3732-3-7 occurred by selecting for KmR transconjugants and supplying transposase in cis from a linked IS50R element. A frameshift mutation in the transposase gene of the IS50L-tox element was also constructed to decrease the likelihood that suppression or a spontaneous reversion at the UAA (ochre) termination codon of IS50L would create an active transposase. The inability of IS50L-tox to transpose further minimizes the potential for horizontal gene transfer of the tox gene to other bacterial species. Expression of the Tox protein in strains 112-12 and Ps3732-3-7 was demonstrated by an immunological assay (Western blot) and toxicity against larvae of the tobacco hornworm (Manduca sexta). PMID- 3036647 TI - Complete nucleotide sequence of the mouse lactate dehydrogenase-A functional gene: comparison of the exon-intron organization of dehydrogenase genes. AB - The complete sequence of 12,851 nucleotides of the mouse lactate dehydrogenase-A (LDH-A) gene has been determined. It includes eight exons, seven introns, promoter and regulatory regions. The B1 repetitive elements present in intron III and VI are oriented in opposite orientation, and they share 72% sequence homology. The exon-intron organization of mouse LDH-A gene is compared with the organizations of other dehydrogenase genes, and the molecular evolution of the nicotinamide adenine dinucleotide binding domains is discussed. PMID- 3036649 TI - Plasmid Reference Center Registry of transposon(Tn) and insertion sequence (IS) allocations through December 1986. AB - The current entries of transposable genetic elements (both Tn and IS) filed with the Plasmid Reference Center are tabulated. These include Tn entries 3601-4550 [entries 1-3600 are listed in Lederberg, Gene 16 (1981) 59-61 and 18 (1982) 366] and all the filed IS entries. PMID- 3036650 TI - Cloning and nucleotide sequence of different iso-IS231 elements and their structural association with the Tn4430 transposon in Bacillus thuringiensis. AB - A family of five repetitive sequences (RS) has been isolated from a plasmid DNA library of Bacillus thuringiensis strain berliner 1715. In a previous paper [Mahillon et al., EMBO J. 4(1985)3895-3899] one of these was shown to harbor all the features of an IS element (IS231). Further nucleotide sequence analysis revealed that two other RS, flanking the delta-endotoxin gene, are actually variants of IS231. Comparison of the nucleotide sequences surrounding the iso IS231 elements showed a unique structural association between some of these elements and the transposon Tn4430. Although these IS231 elements have transposed into Tn4430, both these IS231 s and the transposon Tn4430 remain structurally intact. PMID- 3036651 TI - Amplifiable expression vectors for mammalian cell lines. AB - We have constructed a prototype of a general expression vector for mammalian cells, which carriers a strong promoter active in a variety of tissues and animal species for expression of the gene of interest and a truncated gene for amplification in selective medium. Expression of the S gene of HBV encoding the surface antigen was used to evaluate various versions of this vector. The human metallothionein IIA promoter was found to be particularly efficient for expression of this gene, both in mouse L and monkey Vero cells. Including a promoterless tk gene in the vector led to gene amplification in Ltk- cells by selection of TK+ variants in selective medium containing hypoxanthine, aminopterine and thymidine (HAT medium). Concomitant increases of the S gene expression levels were initially 50-100 fold. Although many clones were unstable, even in selective medium, some maintained the high expression levels for over one year. PMID- 3036652 TI - Cloning and characterization of the aldA gene of Aspergillus nidulans. AB - We have cloned and sequenced the aldA (encoding aldehyde dehydrogenase) gene of Aspergillus nidulans. The gene contains two introns which are similar in size and structure to other fungal introns. The amino acid sequence of aldehyde dehydrogenase (497 residues) shows a significant level of homology with analogous sequences in other organisms. Comparison of the primary structure of the active sites of the mammalian cytosolic and mitochondrial enzymes shows that the Aspergillus enzyme closely resembles the mammalian mitochondrial enzyme. Analysis of the 5' non-coding region of the aldA gene shows a TATA-like sequence located 90 bp upstream from the initiation codon. Two messenger-RNA start points are located 36 and 42 bp upstream from the start codon. PMID- 3036653 TI - Isolation of a mouse heat-shock gene (hsp68) by recombinational screening. AB - We have used cloned fragments from a Drosophila melanogaster hsp70 gene and a mouse hsp68 cDNA in recombinational screens of mouse genomic libraries. Using the mouse probe we have isolated two overlapping recombinant lambda phages comprising 22 kb of cloned DNA. Southern analysis has localized the homology with the Drosophila hsp70 coding region to a 2.2-kb fragment containing the mouse heat shock gene. Insertion accompanying recombinational screening can disrupt interesting sequences; we have overcome this inconvenience by developing a simple one-step genetic selection for phage which have precisely excised the microplasmid probe. PMID- 3036654 TI - Vectors with restriction site banks. V. pJRD215, a wide-host-range cosmid vector with multiple cloning sites. AB - The construction of a new wide-host-range, restriction-site bank, cosmid-cloning vehicle (pJRD215) is described. The wide-host-range properties and the ability to be transferred by conjugation, extend genetic engineering to those Gram-negative species that cannot be transformed. The vector permits the cloning of genes from Gram-negative bacteria using a complementation screening procedure in a mutant host. This procedure is simplified by the possibility of construction of a cosmid gene bank so that only a few hundred clones need to be screened. Subsequent subcloning of the gene of interest is facilitated by the presence of at least 23 unique cloning sites. PMID- 3036655 TI - Regulation of expression of the cloned repE gene from the F plasmid of Escherichia coli. AB - The repE gene of the Escherichia coli F plasmid has been fused to an N-terminal fragment of the Salmonella typhimurium araB gene in the plasmid expression vector pING1. A fusion protein is expressed at high levels upon addition of arabinose to E. coli hosts containing the recombinant plasmid and has been purified to homogeneity. Antibodies prepared against the fusion protein react with both araB' and repE-coded proteins and can be used to detect their synthesis by immunoblotting methods. In vitro as well as in vivo expression of the repE gene from chimeric plasmids indicate a very tight control of the expression of this replication initiator protein. PMID- 3036656 TI - Use of a phage vector for rapid synthesis and cloning of single-stranded cDNA. AB - We have developed a technique for synthesis of single stranded complementary DNA (ss cDNA) using specifically designed phage ssDNA as vector primer. This vector (pPBS27) was constructed by introducing a poly(dT) tail adjacent to the XbaI site of pTZ18R, which can exist either as a plasmid in Escherichia coli or as a ssDNA phage. The pPBS27 phage vector is linearized with XbaI using a restriction-site directed fragment and used to anneal a mixture of poly(A) + RNA for cDNA synthesis by reverse transcriptase. The RNA is then hydrolysed with NaOH and a poly(dG) tail added to the 3' end of the vector-cDNA with terminal transferase. The linear hybrid ssDNA is then closed by annealing with a 15-mer site-directed fragment oligodeoxynucleotide molecule and ligated with T4 DNA ligase. Almost 10(5) E. coli transformants per microgram of vector primer can be obtained in two days. PMID- 3036657 TI - Phycocyanin alpha-subunit gene of Anacystis nidulans R2: cloning, nucleotide sequencing and expression in Escherichia coli. AB - The cloning and nucleotide sequence determination of the Anacystis nidulans R2 phycocyanin (PC) alpha-subunit gene are described. A 3.0-kb PstI fragment of Anacystis nidulans R2 genomic DNA cloned in plasmid pUC8 was found to hybridize with a heptadecameric oligodeoxynucleotide probe. Sequencing using synthetic primers revealed the presence of the PC alpha-subunit gene and the 3' proximal end of the beta-subunit gene. The alpha-gene is separated from the upstream beta gene by a spacer length of 51 bp. The deduced amino acid (aa) sequence of the alpha-subunit protein is identical, except for 5 aa, to that of A. nidulans 6301 and is highly homologous (77%) to that reported for Agmenellum quadruplicatum PR6. The 16-kDa alpha-subunit protein, detected by immunoadsorption, was fortuitously expressed in Escherichia coli from the lacZ promoter of the cloning vehicle pUC8. PMID- 3036659 TI - Sequence and expression characteristics of a shuttle chloramphenicol-resistance marker for Saccharomyces cerevisiae and Escherichia coli. AB - An efficiently transforming chloramphenicol-resistance (CmR) shuttle marker for Saccharomyces cerevisiae and Escherichia coli has been characterized in terms of its primary structure and expression characteristics. The complete nucleotide (nt) sequence of the CmR marker is given, with details on restriction sites, apparent expression signals for both organisms, and translation of the Cm acetyltransferase (CAT)-coding sequence. SDS-polyacrylamide gel electrophoresis and Western blotting have confirmed that the marker produced an identical CAT protein in yeast and E. coli. Each copy of the marker, whether present in multiple copies or as a single copy, gave rise to approx. 0.1% of the total soluble protein as CAT in haploid yeast cells. When compared with homologous expression of alcohol dehydrogenase (ADH-I) by the same ADC1 promoter, this represents a 27-fold reduction for CAT expression, which is typical of heterologous gene expression in yeast. When the marker was on a multicopy plasmid in yeast, up to 2.1% of the total soluble cell protein was produced as CAT, but this did not adversely affect the growth of host cells. Increase of the Cm concentration in the medium did not result in an increase in the number of plasmids nor the amount of CAT protein produced, showing that plasmid copy number and marker expression are regulated independently of the selection pressure. In E. coli, the ADC1 yeast-promoter DNA was found to contain both forwards and backwards promoter activity. The level of expression provided by these promoters was equivalent to that of an average E. coli gene. PMID- 3036658 TI - Serotype-converting bacteriophage D3 of Pseudomonas aeruginosa: vegetative and prophage restriction maps. AB - D3 is a temperate serotype-converting bacteriophage of Pseudomonas aeruginosa. A restriction map, based upon BamHI, PstI, PvuI, HindIII and SmaI sites, indicates that the phage genome is 56.4 kb long, and that it possesses cohesive ends. The prophage map suggests a unique insertion site in the strain AK1380 genome. Phage DNA integration occurs upon the circularization of D3 genome with the integration point approximately equidistant from the two ends. PMID- 3036660 TI - Molecular characterization of human immunodeficiency virus from Zaire: nucleotide sequence analysis identifies conserved and variable domains in the envelope gene. AB - To examine the genetic relatedness of human immunodeficiency viruses (HIV) from different geographic locations, we molecularly cloned the genome of HIV isolated from a Zairian AIDS patient. Restriction mapping of the recombinant clone, designated HIV-Zr6, revealed both common (as observed in other HIV isolates) and unique restriction sites. The DNA clone of HIV-Zr6, shown to give rise to infectious cytopathic virus after transfection of cultured lymphoid cells, was sequenced in several regions. The long terminal repeat (LTR), open reading frame 1 (ORF1), C-terminal envelope (env) gene domain, and ORF2 showed less than 6% difference in nucleotide sequence when compared to other HIV isolates including human T-lymphotropic virus-type III (HTLV-III) clone B10, lymphadenopathy associated virus-1 (LAV-1), and AIDS-associated retrovirus-2 (ARV-2). About 15% difference in nucleotide sequences was noted in the N-terminal env gene domain. Alignments of env gene sequences revealed conserved, moderately variable, and hypervariable stretches in the predicted amino acid sequences. This model provides a basis for assessing the significance of sequence variation on properties controlled by the viral Env glycoproteins such as cell tropism and immunogenicity. PMID- 3036662 TI - High-efficiency preparation of DNA from limiting quantities of eukaryotic cells for hybridization analysis. AB - This report describes a DNA preparation method which allows the detection of single-copy genes in samples of as few as 6000 eukaryotic cells. The technique uses proteinase K digestion in detergent and low-gelation-temperature agarose followed by solidification of the agarose and removal of the detergent by diffusion. RNase and restriction enzyme digestion are carried out in solution after remelting the agarose. The procedure can be performed successfully with mammalian cells in suspension, with parasitic protozoa and with pieces of mammalian tissue weighing less than 1 mg. Numerous samples can be processed simultaneously using frozen as well as fresh material. PMID- 3036661 TI - Hybrid protein thymidine kinase gene fusions: plasmid vectors for the study of transcription and translation initiation signals. AB - The thymidine kinase (TK) gene (tk) from Herpes simplex virus type 1 has been used to form gene fusions encoding enzymatically active hybrid proteins. The promoter, translation initiation region, and the first three codons of the tk gene were removed and replaced with a series of DNA restriction sites. DNA fragments containing gene initiation regions were cloned into these sites and shown to synthesize enzymatically active proteins in Escherichia coli. These gene fusions were shown to complement an E. coli strain which is deficient in TK function. Gene initiation regions were used from the lac operon, the tnpR gene of Tn3, and the insA gene of ISl. TK synthesis was regulated by the control signals of the promoter fused to tk, and was dependent upon the phase alignment of the codons at the fusion joint. The size of the resulting protein was shown to be increased over the size of the original TK protein by the length of the coding region fused to TK. This demonstrated that the tk gene has non-essential N terminal amino acids that can be replaced by other amino acid sequences with the retention of TK enzymatic activity. Such tk gene fusions are useful in situations where fusions with other genes cannot be conveniently selected or assayed. PMID- 3036663 TI - A transposable promoter and transposable promoter probes derived from Tn1721. AB - We describe two types of new Tn1721-derivatives capable of random insertion and of generating transcriptional fusions at the site of insertion: transposable promoters (Tn1735) carrying a strong, inducible ptac promoter that turns on adjacent (cryptic) genes; and transposable promoter probes (Tn1736, Tn1737) carrying promoterless genes coding for chloramphenicol acetyl transferase or beta galactosidase, and used to accurately determine the expression of external promoters. These elements are available with four different selectable resistance markers and on conjugative, temperature-sensitive and multicopy plasmid vehicles. Experiments are described that demonstrate the advantage of random insertions for expressing various genes and for studying gene regulation. PMID- 3036664 TI - SV40-based Escherichia coli shuttle vectors infectious for monkey cells. AB - We describe SV40-based Escherichia coli shuttle vectors which can be packaged as pseudovirions without excision of plasmid sequences and which can be rescued in bacteria. These vectors replicate and are transmitted as virus in monkey COS cells without requiring a helper virus. Extrachromosomal vector DNA isolated from infected cells can be rescued in E. coli, so that DNA alterations can be easily screened. Indeed, some of the constructions give rise to very stable plasmids with no detectable rearrangements after multiple lytic cycles in COS cells. The spontaneous mutation frequency measured in bacteria, on the lacO target, is smaller than those usually found with shuttle vectors. We also constructed an expression vector derived from one of our infectious viruses by inserting the chloramphenicol acetyl transferase gene, expressed from the SV40 early promoter, which is efficiently transduced to cells by infection. In this system, the shuttle virus combines the convenience of plasmid rescue and analysis in bacteria, with the advantages of infectious virus. PMID- 3036665 TI - Extracellular proteins of Vibrio cholerae: molecular cloning, nucleotide sequence and characterization of the deoxyribonuclease (DNase) together with its periplasmic localization in Escherichia coli K-12. AB - The gene encoding the extracellular DNase of Vibrio cholerae was cloned into Escherichia coli K-12. A maximal coding region of 1.2 kb and a minimal region of 0.6 kb were determined by transposon mutagenesis and deletion analysis. The nucleotide sequence of this region contained a single open reading frame of 690 bp corresponding to a protein of Mr 26,389 with a typical N-terminal signal sequence of 18 aa which, when removed, would give a mature protein of Mr 24,163. This is in good agreement with the size of 24 kDa, calculated directly by Coomassie blue staining following sodium dodecyl sulphate-polyacrylamide gel electrophoresis and indirectly via a DNA-hydrolysis assay. The protein is located in the periplasmic space of E. coli K-12 unlike in V. cholerae where it is excreted into the extracellular medium. The introduction of the DNase gene into a periplasmic (tolA) leaky mutant of E. coli K-12 facilitates the release of the protein, further confirming the periplasmic location. PMID- 3036666 TI - On the role of repeated sequences 5' to variant surface glycoprotein genes in African trypanosomes. AB - In African trypanosomes, the DNA region situated upstream from all active and some silent variant surface glycoprotein genes (VSG genes) has a repetitive structure. This region is composed of a variable number of tandem repeats of an A + T-rich sequence which lacks the recognition sites for most commonly used restriction endonucleases, and is thus called 'barren region'. The length of the barren regions varies in different trypanosome variants from 0.2 to many kb. We have characterized the barren region upstream from the active VSG gene in two independent Trypanosoma equiperdum variants expressing the same VSG gene in the same expression site. To analyse the junction point between the expression site and the inserted gene, these two barren regions were cloned and sequenced. The longer barren region contains 14 repeats and the other contains two repeats. In both cases the junction point has been shown to lie within a repeat but different repeats were used in each case. These results argue that the repeats are important for the insertion of the duplicated-transposed gene into the expression site and that any repeat can be used. PMID- 3036667 TI - The nucleotide sequence of the amdS gene of Aspergillus nidulans and the molecular characterization of 5' mutations. AB - The structure of the amdS gene of Aspergillus nidulans has been determined by nucleotide sequence analysis. The coding sequence is interrupted by three small introns with splicing signals consistent with other fungal genes. Possible TATA and CAAT elements are found upstream of the start point of transcription. Sequence changes in mutations in the 5' region of the gene have been determined. Two deletions including the start point of transcription abolish detectable transcripts. A series of mutations in the first exon cause frame shifts leading to polypeptide chain termination in the N-terminal region of the polypeptide. Levels of amdS RNA are reduced in these mutants indicating that translational termination can lead to decreased transcription or (more probably) to lower mRNA stability. The amdI18 mutation has properties consistent with it having a general up-promoter effect on amdS expression and results from a single base pair substitution approx. 77 bp upstream from the transcription start point. PMID- 3036669 TI - [Hygienic characteristics of working conditions of field-crop cultivation in relation to the use of agrochemicals]. PMID- 3036668 TI - Cloning and nucleotide sequence of a carbomycin-resistance gene from Streptomyces thermotolerans. AB - Two plasmids (pOJ158 and pOJ159) containing DNA fragments from the carbomycin(Cb) producing strain Streptomyces thermotolerans were identified in Streptomyces griseofuscus based on their ability to confer resistance to Cb. The Cb-resistance determinants on pOJ158 and pOJ159 were designated carA and carB, respectively. In S. griseofuscus, pOJ159 also confers resistance to spiramycin, rosaramicin, lincomycin, and vernamycin B, but not to tylosin; in Streptomyces lividans, pOJ159 additionally confers resistance to erythromycin and oleandomycin. The carB gene was localized on pOJ159 to a 1.25-kb region whose nucleotide sequence was determined. The sequence has a G + C content of 68% and contains the coding sequence for carB and portions of the 5' and 3' untranslated regions. A comparison of the amino acid sequence of the protein encoded by carB (as deduced from the nucleotide sequence) with the deduced amino acid sequence of the RNA methylase from Streptomyces erythraeus (encoded by ermE) revealed extensive homology, suggesting that carB also encodes an RNA methylase. The region 5' to the coding sequence does not contain a small ORF or regions of complementarity that are commonly associated with translationally regulated macrolide-lincosamide streptogramin B resistance genes. The 3' untranslated region contains an inverted repeat sequence that potentially can form a stable RNA stem-loop structure with a calculated delta G of -70 kcal. PMID- 3036670 TI - [Hygienic evaluation of industrial by-products used as mineral fertilizers]. PMID- 3036672 TI - Platelet membrane components and receptors. AB - Receptor-ligand interactions on the platelet membrane control a number of phenomena unique to platelets such as shape change, aggregation, release reaction of secretion, clot retraction and activation of clotting factors. Localization of receptor on the well-defined platelet membrane glycoproteins appears to be a major target of numerous studies in the field of platelet research. The author is reviewing methodological development in this field including surface labeling of the platelets, one and two dimensional electrophoresis, development of monoclonal and polyclonal antibodies to platelet membrane glycoproteins and receptors, immunoblotting, radiolabeled ligand studies and preparative techniques. Classification of membrane glycoproteins is briefly discussed. The current knowledge of the receptors for fibrinogen and other adhesive proteins, clotting factors, ADP and prostanoids is reviewed. PMID- 3036674 TI - [The EBV virus: infection, multiplication and carcinogenesis]. PMID- 3036671 TI - Diagnostic reliability of simultaneous measurements of beta human chorionic gonadotropin and pregnancy-specific beta-1-glycoprotein in serum of patients with trophoblastic disease. AB - Serum levels for beta-human chorionic gonadotropin (beta-hCG) and pregnancy specific beta 1-glycoprotein (SP1) in patients with trophoblastic disease were measured by radioimmunoassay and enzyme-linked immunosorbent assay. The beta hCG:SP1 ratios were below 1.0 in all 22 cases of complete hydatidiform mole and in 8 of 9 cases of partial hydatidiform mole. Two (10.5%) of 19 cases of invasive mole involving metastasis had ratios that rose above 1.0 during chemotherapy. Ratios ranged from 1.6 to 29 in 11 of 15 cases of choriocarcinoma before chemotherapy. The remaining 4 cases, diagnosed within 3 months of antecedent pregnancy, had ratios below 0.99. Thus, the difference between choriocarcinoma and nonchoriocarcinoma beta-hCG:SP1 ratios may be due to trophoblastic differentiation based on the developmental stage and with trophoblast age, or due to the mass and potential activity of trophoblastic cells. PMID- 3036673 TI - [Clinical and electroneurographic results following surgery of carpal tunnel syndrome]. AB - The surgical treatment of carpal tunnel syndrome is successful in most cases. Sometimes, however, it is not possible to reach a satisfying result. In order to find out the reasons for failure, we examined 80 surgically treated patients. Clinical and electroneurographical parameters were observed. We could show, that the time of intervention and the age of the patient are responsible for the results. The skin incision has to be planned carefully in order to avoid transection of the palmar cutaneous nerve. In cases of systemic diseases the patient has to be informed preoperatively about the possibility of incomplete recovery. PMID- 3036675 TI - The nature and the occurrence of birefringent material in different organs in fatal drug addiction. AB - Insoluble birefringent tablet filler materials commonly found in tablets used in solution by drug addicts as intravenous injections were investigated microscopically. The following filler materials were investigated: talc, potato- and maize-starch, microcrystalline cellulose, magnesium stearate and siliciumoxid. The morphological characteristics of the different materials are described. Tissue sections (lung, liver, spleen, heart, bone-marrow, kidney, lymph-nodes and endocrine glands) from 33 consecutive fatality cases of intravenous drug addicts autopsied at the University Institute of Forensic Medicine in Copenhagen were studied with special reference to the occurrence and nature of birefringent material. Birefringent material was most often demonstrated in lung tissue (94%), followed by spleen (76%), liver (55%), lymph nodes (portal: 39%) and bone-marrow (24%). The material was always localized intracellularly. Granulomatous reaction was only seen in the lungs. Except for one case, talc was the only foreign material seen in other organs than the lungs, undoubtedly due to its smaller size. The presence of insoluble foreign material in lung tissue of drug addicts indicates a habit of intravenous administration and the amount of the material indicates whether the addict usually injects tablets or only does so occasionally. The presence of birefringent material in the organs have only rarely any obvious clinical implications. PMID- 3036676 TI - [Histochemical detection of neutral proteinases (PML gelatinase) from neutrophilic granulocytes in tear fluid]. PMID- 3036678 TI - gamma-Aminobutyric acid (GABA) and hepatic encephalopathy: testing the validity of electroencephalographic evidence of the GABA hypothesis. AB - Some GABA-ergic drugs are known to produce electroencephalographic and behavioral signs of inhibition as well as alterations of the visual evoked potential (VEP) similar in shape and parameters to those in experimental fulminant hepatic failure. These findings, along with a number of biochemical findings, were considered to support the theory attributing hepatic encephalopathy to enhanced GABA-ergic transmission. The present review demonstrates that it is uncertain whether electrographic correlates of experimental encephalopathy can be attributed to faulty GABA-ergic mechanisms; nor it is clear whether drug-induced alterations of electrocortical potentials and those recorded in acute hepatic failure share a common pathogenesis. Special emphasis is put on findings with visual evoked potentials altered by drugs acting at GABA/benzodiazepine receptors to show the diversity of changes associated with the activity of the GABA system. Attention is called to the fact that brain GABA increase as a mechanism of hepatic encephalopathy was discussed without specifying the nature and anatomical location of allegedly impaired neural circuits utilizing the neurotransmitter. PMID- 3036677 TI - [Omega-3-fatty acids. A new strategy for preventing cardiovascular diseases and lipid metabolism disorders]. PMID- 3036680 TI - Pedunculated hepatocellular carcinoma. Is it an entity? AB - Two cases of pedunculated hepatocellular carcinoma are reported. One of these was known to have been present for 5 years. Both patients are alive 1 and 2 years after surgery though the former has developed a solitary bone metastasis. The literature on 30 previously published cases is reviewed and it is concluded that, minor differences apart, this tumour is not substantially different from hepatocellular carcinomas in general. The slow growth and good prognosis relate to its extrahepatic location which may be explained by an origin from accessory lobes of the liver. PMID- 3036679 TI - Thyroid hormone action at the cellular level. AB - Thyroid hormones influence numerous physiological and biochemical functions. The expression of the hormonal effects involves several events. The interaction of T3 with nuclear receptors, and the stimulation of mRNA production appears to be a major step. Extranuclear binding of thyroid hormones could account for early responses. Plasma membrane receptors may play a role in the cellular uptake of T3 and the stimulation of amino acids and sugar transport. A direct control of oxidative phosphorylation through binding of T3 to mitochondrial binding sites has been proposed. The role of cytosolic binding proteins remains unclear. The understanding of the mode of action of thyroid hormones requires a better knowledge of the molecular events occurring at the nuclear level, and the relation between the nuclear and extranuclear binding sites in the hormonal expression. PMID- 3036681 TI - Primary malignant fibrous histiocytoma of the liver--a case report with immunocytochemical observations. AB - A case of a primary sarcomatous tumour in the liver of an elderly female is reported. The tumour consisted of bundles of spindle cells focally in a storiform pattern, intermingled with bizarre giant cells. Immunocytochemically, carcinoembryonic antigen, alpha-fetoprotein, keratin, desmin and type IV collagen could not be demonstrated. Most tumour cells, however, expressed vimentin, whereas a granular cytoplasmic immunoreactivity for alpha-1-antitrypsin and alpha 1-antichymotrypsin was shown in the giant cells. Ultrastructurally the tumour cells did not show any characteristics of epithelial derivation. The morphological and immunocytochemical data justify the conclusion that the tumour should be classified as a malignant fibrous histiocytoma. PMID- 3036682 TI - Malignant fibrous histiocytoma? PMID- 3036683 TI - Time-limited psychiatric hospitalization of children: a model and three-year outcome. AB - Current emphasis on reducing length of hospital stay has stimulated a review of the philosophical, clinical, and administrative approaches to brief psychiatric hospitalization of children. A model of time-limited hospitalization on an eight bed children's unit was designed to stabilize the patients and triage them back to appropriate levels of community aftercare within 28 days. Clinical and administrative strategies used to facilitate this process included a community based case manager for each patient, focused clinical intervention, and strong parent involvement. The treatment model was generally successful in meeting the goals of hospitalization. Of 212 admissions over three years, 37 percent of the patients at discharge continued to need complex, multi-modal treatments, and 63 percent were referred for less intensive outpatient care. PMID- 3036684 TI - Current pathogenetic concepts of diffuse malignant mesothelioma. PMID- 3036685 TI - Gene organization of haplotypes expressing two different C4A allotypes. AB - The gene organization of C4 haplotypes expressing two different C4A allotypes with a C4B null allele (C4A3A2BQ0 and C4A3A6BQO) was studied using Southern blot analysis with cDNA probes and restriction enzymes which give C4A and C4B locus specific restriction fragments. These haplotypes were shown to have both a C4A and a C4B locus present, suggesting that the C4B locus expresses a C4A protein. The finding of a 21-OH A and a 21-OH B gene on the C4A3A6BQO haplotype further suggests that this haplotype has the common gene organization C4A, 21-OH A, C4B, 21-OH B. A model explaining C4 null alleles on haplotypes found to have two C4 loci is presented. PMID- 3036686 TI - Submicroscopic duplication of chromosome 21 and trisomy 21 phenotype (Down syndrome). AB - A patient with the phenotype of trisomy 21 (Down syndrome) was found to have a normal karyotype in blood lymphocytes and fibroblasts. Assessment of the chromosome 21 markers SOD1, CBS, ETS2, D21S11, and BCEI showed partial trisomy by duplication of a chromosome segment carrying the SOD1, CBS, and ETS2 loci and flanked by the BCEI and D21S11 loci, which are not duplicated. This submicroscopic duplication at the interface of 21q21 and 21q22.1 reduces to about 2000-3000 kb the critical segment the trisomy of which is responsible for the phenotype of trisomy 21. PMID- 3036687 TI - Human delta-aminolevulinate dehydratase: chromosomal localization to 9q34 by in situ hybridization. AB - The structural gene for human delta-aminolevulinate dehydratase (ALA-D) has been localized to chromosomal region 9q34 by in situ hybridization using a [125I] labeled human delta-aminolevulinate dehydratase cDNA. Of the 150 silver grains analyzed, 25% were localized to chromosome 9q, while 12% and 8% were on chromosomes 1p and 13q, respectively. The single chromosomal region q34 had over 90% of the total grains observed on chromosome 9. In contrast, the grains on chromosomes 1p and 13q were dispersed, consistent with the absence of any human ALA-D pseudogenes. Southern blot analysis of somatic cell hybrids informative for ALA-D (Wang et al. 1985) also was consistent and supported the finding of only one locus for this heme biosynthetic enzyme. PMID- 3036688 TI - An improved method for detecting Y chromosomal DNA. AB - The DNA probe Y97 was derived from a repeat sequence in the human Y centromere, a region which must be present in a mitotically functional Y chromosome. We have demonstrated that Y97, which detects a Y-specific 5.5-kb EcoRI fragment by Southern analysis, is very useful for the molecular detection of small amounts of Y-derived material and represents a significant improvement over previous tests for molecular diagnosis of sex. The male-female difference in hybridization was unequivocal even when only 25 ng of total DNA was used per lane. Furthermore, in mixing experiments the 5.5-kb EcoRI fragment was detectable even when only 5% of the total DNA was male. By increasing hybridization stringency, we have developed a rapid, sensitive, and accurate method to detect Y chromosomal DNA in unrestricted samples. PMID- 3036690 TI - A monoclonal antibody, GR7A4, reacting with the T10 antigen. AB - The monoclonal antibody GR7A4 was produced against a human lymphoblastic leukemia (ALL-B). Biochemical characterization of GR7A4 was carried out with Raji cells in SDS-PAGE studies. GR7A4 precipitated a 43 kd molecule. The tissue distribution and molecular weight were similar to the T10 antigen. Modulation and capping of GR7A4 antigen reduced its binding ability to cells with either GR7A4 or OKT10. However, the cell surface distribution pattern observed was somewhat different from other similar monoclonal antibodies. Thus GR7A4 reacted greatly with pre-B cell lines, Burkitt cell lines, EBV cell lines and activated PHA, ConA and PWM lymphocytes, however reactivity with leukemic cells was very limited. The kinetics of appearance of GR7A4 antigen on PWM blasts show that this molecule seems to represent an intermediate stage in lymphocytic activation. The differences in comparison with other similar MAbs are discussed and correlated with the peculiar discontinous pattern of appearance of this antigen. PMID- 3036689 TI - Human alpha-globin maps to pter-p13.3 in chromosome 16 distal to PGP. AB - Fibroblasts from a fetus with an unbalanced karyotype 46(XY), -16,+(16qter 16p13.3::4q31.1-4qter) were found to possess only one allele at the 3' hypervariable region (3'HVR) close to the alpha-globin locus and two alleles at the PGP locus. This places the alpha-globin locus at the very tip of 16p, distal to PGP. PMID- 3036691 TI - Interaction of puromycin and puromycin aminonucleoside with poly (A)--a proton magnetic resonance investigation. PMID- 3036692 TI - Effect of cAMP on down-regulation of erythrocyte insulin receptor. PMID- 3036693 TI - Measurement of cell-mediated immunity against bovine papilloma virus by lymphoproliferative reactions. AB - To study cellular immunity towards bovine papillomavirus (BPV), calves were infected intradermally with BPV-1, and the cellular immune response was measured by the lymphocyte proliferation assay. Peripheral blood lymphocytes were obtained which in control experiments were highly reactive towards mitogen stimulation. Different batches of BPV-1 were prepared by the use of density gradients. In the first set of experiments, a nonspecific mitogenic effect of the virus preparation was observed. This effect was obviously caused by soluble molecules contained in the virion-free supernatant obtained after ultracentrifugation of the viruses. Subsequently, we obtained highly purified virions which did not share the nonspecific mitogenic effect. The data obtained with these virions showed no responses of untreated control animals but a short-lived in vitro lymphoproliferative response 2 to 3 weeks after infection. This response then disappeared and was followed by a period of non-responsiveness which lasted as long as we tested the animals. PMID- 3036694 TI - Electron microscopic study on the role of delayed hypersensitivity & antibody producing cells in protection against herpes simplex virus in nude mice. PMID- 3036695 TI - Efficacy of prophylactic anti-D immunoglobin injections. PMID- 3036696 TI - Role of dietary fibre in irritable bowel syndrome: a clinical study. PMID- 3036697 TI - The role of contaminants in hexachlorobenzene toxicity. AB - Hexachlorobenzene (HCB) is hepatotoxic, causing a porphyria with an acquired deficiency of uroporphyrinogen decarboxylase. Conflicts have arisen regarding the toxicological significance of impurities in samples of HCB. In female rats, unpurified practical-grade HCB from Eastman Kodak Company was more porphyrogenic than a sample from Aldrich Chemical Company that had been recrystallized from benzene. To determine whether specific inducers of hepatic microsomal cytochrome P1-450, such as 2,3,7,8-tetrachlorodibenzo-para-dioxin (TCDD), contributed to the difference in toxicity of these samples, induction of porphyria and activities of the cytochrome P1-450-dependent mono-oxygenase, ethoxyresorufin-O-deethylase (ERR), were compared in rats receiving HCB (0.2% w/w) or TCDD (10 micrograms/kg initial dose, then 2.5 micrograms/kg weekly). Porphyria occurred in the former but ERR activity was induced about four times more in the latter. The relative potency of crude and purified HCB samples in inducing ERR activity was further evaluated in C57Bl/6 and DBA/2 mice; ERR activity was three times higher in C57Bl/6 than in DBA/2 mice treated with 0.01% w/w HCB. ERR activity was also higher in C57Bl/6 mice treated with impure than in those treated with purified HCB. These results show that undefined impurities can be important determinants of the biological effects of HCB samples and suggest that these effects are partly mediated via the Ah receptor. However, porphyria induced by HCB appears to be independent of HCB impurities and not solely dependent on induction of the cytochrome-P1-450 mono-oxygenases. PMID- 3036699 TI - The environmental fate of hexachlorobenzene. AB - The relative importance of different environmental fate processes for hexachlorobenzene (HCB) can be estimated from measured and estimated values of several physical properties and chemical kinetic and equilibrium constants. Photolysis of HCB in water and hexane is slow, while hydrolysis, oxidation and biotransformation appear to be unimportant. HCB will sorb strongly to soil and sediment but will be rapidly volatilized from water into the atmosphere, where slow photolysis is the dominant loss process. PMID- 3036698 TI - Neurotoxicity of hexachlorobenzene-induced porphyria turcica. PMID- 3036700 TI - Papillary collecting tubule responsiveness to atrial natriuretic factor in Dahl rats. AB - There is evidence that atrial natriuretic factor (ANF) has an action in the inner medullary collecting duct. In addition, the prehypertensive Dahl salt-sensitive (S) rat has an intrinsic tendency toward less natriuresis than the Dahl salt resistant (R) rat has when challenged with ANF. To test the hypothesis that renal papillary collecting tubule cells from prehypertensive S rats might be genetically less responsive to ANF, S and R cells were grown in culture and studied for responsiveness to ANF by measurement of cyclic nucleotide responses. There was a concentration-dependent effect of ANF on renal papillary collecting tubule cell synthesis of intracellular cyclic guanosine 3',5'-monophosphate (cGMP) in both strains. However, the S cells were hyporesponsive compared with the R cells (p less than 0.002, by analysis of variance). Likewise, in response to Na nitroprusside, the S cells were hyporesponsive compared with the R cells as measured by intracellular cGMP accumulation (p less than 0.03, by analysis of variance). Arginine vasopressin stimulated intracellular cAMP equally in both strains. Also, ANF equally enhanced intracellular cGMP in glomerular mesangial cells from S and R rats, indicating possible specificity of the reduced responsiveness to ANF to the distal nephron of S rats. Plasma ANF levels had a slight tendency to be higher in prehypertensive S rats than in R rats (p = 0.088, by t test). These results suggest that the papillary collecting duct of Dahl S and R rats may differ in guanylate cyclase activity. This difference may partially explain the impaired natriuretic responses of S rats and could represent a factor contributing to the development of salt-sensitive hypertension. PMID- 3036702 TI - Endothelium-derived relaxing factor in cultured cells. AB - Many vasoactive agents stimulate release of an endothelium-derived relaxing factor (EDRF). EDRF stimulates cyclic guanosine 3',5'-monophosphate (cGMP) accumulation and relaxation of vascular smooth muscle in a manner similar to that produced by sodium nitroprusside. Endothelium and vascular smooth muscle were isolated from porcine, bovine, and rat thoracic aorta. The capacity of sodium nitroprusside to stimulate cGMP accumulation in cultured bovine, porcine, and rat vascular smooth muscle was found to increase with time in culture to a maximum of 12 to 14 days after plating. In addition, bovine and porcine vascular smooth muscle, but not rat vascular smooth muscle, lost the sodium nitroprusside stimulated cGMP response after the fifth passage. Cultured endothelial cells did not respond to endothelium-dependent vasodilators or sodium nitroprusside with increased cGMP levels. Vascular smooth muscle cells responded only to sodium nitroprusside. Mixed cultures of porcine and bovine endothelium and vascular smooth muscle and bovine endothelium and rat vascular smooth muscle responded to endothelium-dependent vasodilators with increased cGMP levels. Short-term (4 hours) coculture experiments using bovine endothelium grown on microcarriers to assess the need for long-term contact between the two cell types produced similar results. Release of EDRF from bovine endothelium was studied by loading endothelium-covered microcarrier beads into a column superfused with physiological buffer. Treatment of the column with bradykinin, the calcium ionophore A23187, melittin, and arachidonate released EDRF from the column as measured by cGMP changes in denuded aortic rings and vascular smooth muscle cells and by relaxation of rings when bathed in column effluent. The time course of cGMP changes and relaxation were similar and could be reversed by hydroquinone.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3036701 TI - Angiotensin II stimulation of vascular smooth muscle phosphoinositide metabolism. State of the art lecture. AB - Phosphoinositide hydrolysis is an integral step in the activation of vascular smooth muscle by angiotensin II. Sequential phospholipase C-mediated hydrolysis of the polyphosphoinositides and phosphatidylinositol in cultured vascular smooth muscle cells stimulated with angiotensin II results in a coordinated series of biochemical events: a transient formation of inositol trisphosphate associated with calcium mobilization, and a biphasic, sustained formation of diacylglycerol associated with activation of protein kinase C and cytosolic alkalinization. The initial, rapid phase and the sustained phase of the angiotensin II response appear to be differentially controlled. Formation of inositol trisphosphate and mobilization of calcium are attenuated by activation of protein kinase C. Sustained diacylglycerol formation is promoted by cytosolic alkalinization, and appears to require cellular processing of the angiotensin II-receptor complex. Calcium and cyclic guanosine 3',5'-monophosphate do not appear to regulate phospholipase C-mediated phosphoinositide hydrolysis in vascular smooth muscle. Thus, regulation of angiotensin II-stimulated second messenger generation in vascular smooth muscle is complex, perhaps involving protein kinase C activation, changes in intracellular pH, and processing of the angiotensin II-receptor complex. PMID- 3036704 TI - Angiotensin II receptors and angiotensin converting enzyme in the medulla oblongata. AB - Quantitative in vitro autoradiography was used to map angiotensin II (ANG II) receptors and angiotensin converting enzyme (ACE) in sections from rat, rabbit, sheep, and human medulla oblongata and to follow changes in receptor and ACE density after disruption of vagal projections by nodose ganglionectomy in the rat. ANG II receptors and ACE are both concentrated in the nucleus of the solitary tract and dorsal motor nucleus of vagus of the rat, rabbit, sheep, and human. An ANG II receptor-containing band connecting the nucleus of the solitary tract with the dorsolateral medulla was seen in rabbit and human tissue, providing evidence for association of ANG II receptors with vagal afferent fibers. ANG II receptors were found to be concentrated in the rostral and caudal ventrolateral medulla, which corresponded to the region of C1 and A1 catecholamine-containing cell groups in the rabbit. This localization was also evident in rat and human tissue. In all four species, a prominent, ANG II receptor-rich band in the intermediate reticular nucleus was found to connect the ventrolateral medulla and the dorsal vagal complex. In humans and sheep, this band contains puncta that overlie cell bodies. One week after nodose ganglionectomy in the rat, the density of ANG II receptors in the ipsilateral dorsal vagal complex fell markedly. This fall was most prominent in the rostral dorsal motor nucleus of vagus (to 46% of control density) and in the nucleus of the solitary tract (to 56% of control). ACE levels and calcitonin gene-related peptide receptor density were unchanged in both nuclei after ganglionectomy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3036703 TI - Electron spin resonance studies of erythrocytes from spontaneously hypertensive rats and humans with essential hypertension. AB - The purpose of the present study was to investigate erythrocyte membrane abnormalities in hypertension by means of an electron spin resonance and spin label technique. The erythrocytes from spontaneously hypertensive rats (SHR) and humans with untreated essential hypertension were examined and compared with their normotensive counterparts, and electron spin resonance spectra were obtained for a fatty spin-label agent (5-nitroxy stearate) incorporated into the erythrocyte membranes. The value of outer hyperfine splitting (2T' parallel) was significantly higher in erythrocytes of SHR and humans with essential hypertension than in erythrocytes of normotensive controls (at 37 degrees C: SHR, 56.14 +/- 0.51 gauss [G], n = 8; Wistar-Kyoto rats, 52.22 +/- 0.86 G, n = 4, p less than 0.01; humans with essential hypertension, 56.94 +/- 0.27 G, n = 11; normotensive subjects, 55.44 +/- 0.36 G, n = 8, p less than 0.01). The order parameter (S) was also increased in the hypertensive rats and humans compared to their respective normotensive controls. When calcium was loaded to erythrocytes with calcium ionophore A23187 (0.9 microM) and CaCl2 (1.0 mM), the parameters of the spectra were increased. These changes were more prominent in the hypertensive groups than in the normotensive controls. These results revealed that the erythrocyte membranes of the hypertensive subjects tolerated different spin motions than those of the normotensive controls in the electron spin resonance study and that membrane fluidity might be decreased in hypertension. Additionally, calcium loading to erythrocytes caused the reduction of membrane fluidity. Therefore, it is suggested that an abnormality of calcium handling at the cellular level might affect physical properties of the biomembranes in hypertension. PMID- 3036705 TI - Plasma angiotensins and blood pressure during converting enzyme inhibition. AB - The relationship between plasma angiotensin and the reduction of blood pressure with the angiotensin converting enzyme inhibitor enalapril was studied in rats. Blood pressure was measured in conscious rats with indwelling arterial catheters. To measure angiotensin II, plasma was analyzed by physical separation of angiotensins using high performance liquid chromatography followed by radioimmunoassay. The effects of both single (acute) and long-term (chronic) dosages of enalapril were measured. After a single oral dose of enalapril (10 mg/kg), mean arterial pressure fell from 111 +/- 3 to 86 +/- 3 mm Hg (p less than 0.005). Despite the blood pressure reduction, plasma angiotensin II was unaffected (control, 9.9 +/- 1.8 vs 9.7 +/- 1.1 pg/ml). After a higher single oral dose of enalapril (30 mg/kg), there was a reduction in both mean arterial pressure (81 +/- 5 mm Hg, p less than 0.005) and plasma angiotensin II concentration (2.3 +/- 0.6 pg/ml, p less than 0.01). The chronic effects of converting enzyme inhibition were evaluated in rats given enalapril in their drinking water (30 mg/kg 24 hr) for 1 week or 2 months. Mean arterial pressure remained equally low after chronic administration (for 1 week or 2 months), but plasma angiotensin II increased above normal (after 1 week, 28.9 +/- 8.7, p less than 0.01 vs control; after 2 months, 43.1 +/- 16.2 pg/ml, p less than 0.05 vs control). Although plasma angiotensin converting enzyme activity was undetectable at any time after enalapril administration, there was a partial return of the angiotensin I pressor response with chronic administration. The data are most compatible with actions of converting enzyme inhibitors independent of the blockade of plasma angiotensin II formation. PMID- 3036706 TI - Subpressor infusions of angiotensin II alter glomerular binding, prostaglandin E2, and cyclic AMP production. AB - Angiotensin II (ANG II) has been postulated to have pathogenetic role in diminished glomerular function in a number of animal models of acute renal failure. The present studies were designed to test the hypothesis that modest elevations in circulating ANG II potentiate the ability of ANG II to reduce glomerular capillary surface area through an effect on ANG II binding to glomerular mesangial cells and/or influences on other modulators of function. Rat glomeruli isolated by a sieving technique were employed in vitro in an ANG II radioreceptor assay. Subpressor infusion of ANG II for 36 hours in rats increases the affinity and number of ANG II binding sites of isolated glomeruli. The ability of ANG II to influence function was tested by assessing its effect on glomerular surface area in vitro by image-analysis microscopy, a method of measuring mesangial cell contractility. The sensitivity and magnitude of ANG II induced decrements in glomerular surface area were increased. ANG II infusion diminished glomerular prostaglandin E2 (PGE2) production, increased basal cyclic adenosine 3',5'-monophosphate (cAMP) production, and enhanced ANG II-induced decrements in cAMP production. In control glomeruli, only pharmacological concentrations of ANG II inhibited cAMP, but after ANG II infusion, physiological concentrations of ANG II were capable of inhibiting cAMP by as much as 57% (below basal values). In conclusion, continuous infusion of subpressor concentrations of ANG II in rats enhances the contractile response of the glomerular mesangial cell through effects on the cell's surface receptor for ANG II and on prostaglandin and cAMP production. These actions may be important mediators of the effects of ANG II on glomerular function associated with a number of experimental models of kidney disease. PMID- 3036707 TI - Phorbol ester, vascular relaxation, and cyclic guanosine 3',5'-monophosphate. AB - The relaxation of phenylephrine-contracted blood vessels by acetylcholine, nitroprusside, or atrial natriuretic factor has been linked to elevations in cyclic guanosine 3',5'-monophosphate (cGMP). Also, 8-bromo-cGMP can induce vascular relaxation in isolated vascular smooth muscle contracted with phenylephrine. We determined whether these cGMP-dependent vasodilators could relax isolated rat aortas contracted with the phorbol ester 12-O tetradecanoylphorbol-13-acetate. cGMP was measured by radioimmunoassay. Acetylcholine, nitroprusside, and atrial natriuretic factor induced relaxation in vascular smooth muscle contracted by 12-O-tetradecanoylphorbol-13-acetate. These relaxation responses were accompanied by elevations of cGMP. However, the sensitivity to these vasodilators was markedly decreased in phorbol ester contracted vessels compared to phenylephrine-contracted vessels. Nifedipine and superoxide dismutase induced small but significant relaxations in phorbol ester contracted vessels; however, blood vessels contracted with phenylephrine and phorbol ester relaxed completely with papaverine. There was a marked decrease in sensitivity to 8-bromo-cGMP in phorbol ester-treated vessels compared to phenylephrine-contracted vessels. Contractions induced by phorbol ester were not inhibited by amiloride or chlorpromazine. Also, following incubation in potassium free salt solution, vessels incubated with phenylephrine or phenylephrine and phorbol ester underwent similar relaxations when exposed to potassium chloride. The contractile state induced by phorbol ester has decreased sensitivity to cGMP dependent vasodilators. This may be due to nonspecific effects of the phorbol ester or to the mechanism by which protein kinase C activation maintains vascular tone. PMID- 3036708 TI - Evidence for sialyl glycoconjugates as receptors for Bordetella bronchiseptica on swine nasal mucosa. AB - The nature of the receptors for Bordetella bronchiseptica was investigated by using the in vitro adherence assay system. The results indicated that sialyl glycoconjugates acted as receptors on swine nasal mucosa. These results were obtained by two independent approaches: inhibition of epithelial cell adherence with sialic acid-containing compounds but not with compounds lacking sialic acid residues and loss of adherence after treatment of epithelial cells with periodate or neuraminidase. B. bronchiseptica seems to have strong affinity for mucin. This may help the bacterium to colonize the mucosal surfaces of the swine nasal cavity. PMID- 3036710 TI - Biological and biochemical characterization of tunicamycin-resistant Leishmania mexicana: mechanism of drug resistance and virulence. AB - A parasitic protozoan, Leishmania mexicana amazonensis, was previously made resistant to tunicamycin (J.A. Kink and K.-P. Chang, Proc. Natl. Acad. Sci. USA 84:1253-1257, 1987). In the present study, six different tunicamycin-resistant variants were biologically and biochemically compared with their parental wild type to further delineate the mechanism of tunicamycin resistance and that of their virulence observed. In contrast to their parental wild type, all tunicamycin-resistant variants were found to grow and differentiate in tunicamycin-containing medium. The 50% lethal doses of tunicamycin for variants resistant to 10 or 80 micrograms of tunicamycin per ml were 20- and 100-fold higher, respectively, than that of the wild type. Specific activity of the microsomal N-acetylglucosamine-1-phosphate transferase was 4- to 12-fold higher in the tunicamycin-resistant cells than in their parental wild type and tunicamycin-sensitive revertants. The level of the enzyme activity is proportional to the degree of drug resistance. Inhibition kinetics studies showed that the enzyme from all groups was equally sensitive to the drug, with a 50% effective concentration of 1 to 1.3 micrograms of tunicamycin per ml. Thus, tunicamycin resistance of the variants is caused primarily by an increased level of their enzyme without alteration of its structure. Protein glycosylation determined by the incorporation of 2-D-[3H]mannose was about twofold higher in the tunicamycin-resistant variants than in their parental wild type. The increased glycosyltransferase activity in the latter apparently renders their protein glycosylation insensitive to the inhibition by tunicamycin. A major membrane glycoprotein of 63 kilodaltons (gp63) on the leishmania surface was found to be about threefold higher in the tunicamycin-resistant variants than in the wild type, as determined by immunoprecipitation with a monoclonal antibody specific for this antigen. Tunicamycin treatment of the wild type and tunicamycin resistant variants caused changes in the electrophoretic mobility of this molecule, indicating a higher degree of its glycosylation in the latter cells. The tunicamycin-resistant variants parasitized macrophages in vitro more effectively than did the wild type, accounting for their virulence seen in mice. Thus, a high level of the glycosyltransferase enables the tunicamycin-resistant cells not only to overcome the inhibitory effect of tunicamycin on protein glycosylation but also to express their virulence, possibly by regulating N glycosylation of leishmanial proteins critical for leishmanias to establish intracellular parasitism. PMID- 3036711 TI - Isolation and restriction site maps of the genes encoding five Mycobacterium tuberculosis proteins. AB - A series of recombinant phage expressing five Mycobacterium tuberculosis antigens were isolated. Restriction maps for these antigens were deduced, and the size of the expressed proteins was determined. PMID- 3036712 TI - Etiological, clinical and laboratory data of post-transfusion hepatitis: a retrospective study of 379 cases from 53 Italian hospitals. AB - Among the 8,604 cases of acute viral hepatitis hospitalized during 1982 in 53 Italian hospitals, we studied 379 cases of post-transfusion hepatitis, 262 cases which occurred after surgery and 4,576 cases with no history of parenteral exposure. The etiological agents of post-transfusion hepatitis were NANB viruses in 57.8%, HBV in 39.0% and HAV in 3.2% of the cases. CMV and EBV accounted for less than 1.5% of the post-transfusion hepatitis cases. HBV was the main etiological agent (62.2% of the cases) in the post-surgical hepatitis group, where HAV accounted for only 6.1% of the cases. In contrast, in the group with no history of parenteral exposure, hepatitis A was most frequent. Percentages of patients with history of transfusion or surgery were always higher in type B and NANB hepatitis than in type A, suggesting that surgery without transfusion also represents a risk of acquiring type B and NANB hepatitis. No regional differences were observed in the etiological patterns of post-transfusion hepatitis and post surgical hepatitis. The acute phase of type B post-transfusion hepatitis was more severe than that of NANB post-transfusion hepatitis, as shown by higher serum bilirubin and ALT levels and by a higher case fatality rate. PMID- 3036709 TI - Nonoxidative microbicidal activity in normal human alveolar and peritoneal macrophages. AB - Although Toxoplasma gondii multiplies within normal murine alveolar and peritoneal macrophages, it is killed by normal rat alveolar and peritoneal macrophages. The killing by rat macrophages is by a nonoxidative mechanism. Studies on normal human alveolar macrophages have reported disparate results in regard to their ability to inhibit or kill T. gondii. We considered it of interest to explore further the effect of normal human alveolar and peritoneal macrophages on T. gondii. Unstimulated alveolar macrophages from each of seven individuals demonstrated a marked ability to kill or inhibit multiplication of T. gondii in vitro (e.g., the number of parasites per 100 alveolar macrophages was 31 at time zero and 2 at 18 h, whereas this value increased from 37 at time zero to 183 at 18 h in murine macrophages assayed in parallel). In quantitative assays of superoxide, alveolar macrophages released a substantial amount of superoxide when exposed to phorbol myristate acetate or to candidae. In contrast, alveolar macrophages incubated with T. gondii released no more superoxide than when in medium alone. Scavengers of superoxide anions, hydrogen peroxide, singlet oxygen, and hydroxyl radicals failed to inhibit killing of T. gondii by alveolar macrophages. Peritoneal macrophages from each of six normal women undergoing laparoscopy killed T. gondii in vitro; results of quantitative superoxide assays and scavenger experiments demonstrated that no oxidative burst was triggered in these macrophages by exposure to T. gondii. These data indicate that normal human alveolar and peritoneal macrophages can kill an intracellular parasite by nonoxidative mechanisms and suggest that these mechanisms are important in inhibition or killing of other opportunistic intracellular pathogens. PMID- 3036713 TI - Hamster model for herpes simplex virus infection of the central nervous system. AB - Adult Syrian hamsters were inoculated with a mouse brain passage of herpes simplex virus type 1 (HSV-1) along an intradermal and an oral route after local scarification. For both routes, clinical symptoms of central nervous system (CNS) involvement were seen in the period between five and 12 days post infection. Compared with the route via the buccal mucosa, CNS involvement by intradermal infection is easier to establish. With a minimum dose of 2 X 10(4) TCID50 virus via the intradermal route, an infection rate of 80% or more can be obtained reproducibly and detected simply by clinical observation without need of a survey of the humoral immune response. Scarification of the skin prior to inoculation was found to be essential for establishment of the CNS infection. PMID- 3036714 TI - Serologic detection of active infections with human herpes viruses (CMV, EBV, HSV, VZV): diagnostic potential of IgA class and IgG subclass-specific antibodies. AB - In 175 sera from healthy persons as well as those suffering from primary or secondary herpes virus infections/reactivations, serum antibodies were assessed by an indirect ELISA in the immunoglobulin classes A, G and M and the subclasses G1-4, using carrier-fixed antigens (CMV, VZV, HSV) and monoclonal tracer antibodies. In a similar way EBV-specific antibodies were tested by an indirect IFT. Only IgG1 antibodies were detectable in nearly all persons. Virus-specific IgA and IgG3 may support conventional serological methods (IgM, IgG) indicating recent infection/reactivation with VZV, EBV and possibly CMV. Furthermore, differentiation of primary and secondary CMV and VZV infection was possible in some cases, when IgG3 was detectable before IgG1 in subsequent blood specimens. Recurrent herpes lesions could not be diagnosed serologically. PMID- 3036715 TI - Acyclovir treatment in infectious mononucleosis: a clinical and virological study. AB - Fifty-six patients with a clinical and laboratory diagnosis of infectious mononucleosis who had not been ill for more than seven days, were randomised for peroral treatment with acyclovir (800 mg five times daily) or placebo for seven days in a double blind trial. Clinical, virological and immunological parameters were monitored in each patient for six months. During treatment, shedding of Epstein-Barr virus' as assessed in 36 patients, was significantly reduced (p less than 0.001). However, virus production in the oropharynx returned to pre treatment levels one week after the cessation of therapy. Virus was detected in 35 patients at enrollment and in 28 of 36 patients at the six-month control. No effect on the clinical course of the disease was noticed. The virus-specific antibody response was also unaffected. A significant reduction in spontaneous outgrowth of in vivo Epstein-Barr virus-infected B-lymphocytes was found at 180 days after treatment in four acyclovir-treated patients compared to six controls (p less than 0.001). In another three patients with over-whelming clinical symptoms causing airway obstruction and/or disseminated intravascular coagulopathy, treatment with intravenous acyclovir (10 mg/kg three times daily) was combined with prednisolone (0.7 mg/kg daily) for ten days. Virus shedding ceased transiently during treatment, but returned to initial levels within one week. A dramatic clinical effect on the pharyngeal oedema and general health of the two patients with airway obstruction was noticed, but was much less evident in a patient with intravascular coagulopathy. PMID- 3036716 TI - Management of viral infections in AIDS patients. AB - Viral infections, predominantly those of the herpes virus family, account for up to 16% of all clinically significant infections in AIDS patients. Acyclovir has provided successful treatment in AIDS patients suffering from severe herpes simplex and herpes zoster virus infections. Preliminary results are presented on newly developed acyclovir analogues. Desciclovir, an oral prodrug of acyclovir which is metabolized to acyclovir in vivo, allows treatment of virus infections per os, where high serum levels are needed, e.g. in Epstein-Barr virus infections. BW B759U, another analogue of acyclovir, has been used for the treatment of life-threatening or sight-threatening cytomegalovirus infections in AIDS patients. More than 80% of the patients treated for retinitis experienced stabilization or clinical improvement. Antiviral efficacy was demonstrated in 73% of the patients. Azidothymidine, a nucleoside analogue of thymidine, has been developed specifically to treat the HIV infection. Its antiviral activity is based on inhibition of reverse transcriptase. Phase I studies have demonstrated that azidothymidine is well tolerated. Its ability to cross the blood brain barrier makes it an attractive candidate for treatment of HIV. Trials to determine efficacy are in progress. PMID- 3036717 TI - A mammalian cell line designed to test the mutagenic activity of anti-herpes nucleosides. AB - The herpes simplex virus (HSV) thymidine kinase (tk) gene was transfected into Chinese hamster ovary (CHO) 51-11 gly- cells to test its effect on the cytotoxic and mutagenic activity of anti-herpetic nucleoside analogues. Insertion of the viral tk was verified by Southern blot analysis, by sensitivity to acyclovir, and by elevated in vitro thymidine kinase (TK) activity. TK activity was increased by superinfection with a tk- virus and inhibited by antibody to viral TK. Acyclovir (ACV) was somewhat more cytotoxic in the 51-D3 cell line that expresses the viral TK than in the 51-11 parent line. Growth in ACV did not increase over background mutations at the hprt locus. FIAC (2'-fluoro-5-iodio-aracytosine) was slightly cytotoxic to the parent 51-11 line and the tk-containing clone 51-D3. FMAU (2' fluoro-5-methyl-arauracil) had pronounced cytotoxicity in both cell lines: the 50% survival points were 1.0 microM for 51-11 cells and 0.2 microM for 51-D3. The clone 51-D3 was more sensitive than 51-11 to low concentrations of FIAU (2' fluoro-5-iodo-arauracil), and when treated with FIAU 51-D3 had a mutation frequency to glycine independence 5 times greater than that of 51-11 cells. With both cell lines the mutation frequency at the hprt locus did not increase after growth in the presence of FIAC or FIAU. A 7-fold increase in mutation frequency at the hprt locus was detected after 51-D3 cells were grown with iododeoxyuridine. Trifluorothymidine was more toxic to 51-D3 than to 51-11 cells and increased the mutation frequency 2-fold. Cytosine-beta-D-arabinofuranoside showed no differential cytotoxicity on the two cell lines and did not increase the mutation frequency at the hprt locus. PMID- 3036718 TI - Regulation of tumor development: the biphasic effects of silica and of lipopolysaccharide on natural resistance. AB - The impact on tumor development of the BRM silica and LPS was assessed through analysis of changes in NR parameters in vivo and in vitro. Although injection of the fumed silica Cab-o-sil 3 days before a threshold s.c. inoculum of L5178Y-F9 cells increased the tumor frequency in syngeneic DBA/2 mice, tumors recovered from silica-treated animals exhibited an augmented resistance to NAb and to in vivo NR. Cab-o-sil increased in vivo NR and induced a biphasic modulation of anti tumor NAb and NK activities. The appearance of more autonomous tumors in Cab-o sil-treated mice corresponding with a stimulation of NR parameters, suggests that the adjuvant activity of silica also contributes to its co-carcinogenic effect by accelerating tumor development. While injection of LPS 2-3 days before a threshold tumor inoculum lowered the tumor incidence, the survival of tumor cells injected within 1 day of LPS was increased. A corresponding early decrease in NAb activity occurred, in contrast with increases in NK cell and NAb levels previously observed after 5 days. This biphasic effect of LPS on NR effectors assayed in vitro was also seen on in vivo NR. Although their frequency was higher, tumors initiated during the period of LPS-induced NR abrogation exhibited greater reductions in NAb binding and sensitivity to NR than tumors from control mice. These data extend the support for NAb acting against tumor cells in vivo and reveal the dual nature of NR in tumor development, defending against small tumor foci and driving the progression of the surviving neoplasm. PMID- 3036720 TI - Recombinant feline viruses containing the myc gene rapidly produce clonal tumours expressing T-cell antigen receptor gene transcripts. AB - We, and others, have recently shown that recombinant feline leukaemia viruses (FeLV) containing the myc gene (FeLV-myc) occur in up to 30% of naturally occurring cases of T-cell lymphosarcoma. Investigation of the disease spectrum of two FeLV-myc isolates showed that they induced clonal or oligoclonal T-cell tumours after a short latent period. The phenotypic pattern of the thymic tumours was restricted in that they all expressed the alpha and beta chains of the T-cell antigen receptor and could readily be established in culture in vitro without the addition of exogenous interleukin-2. Although helper FeLV was transmitted from infected cats to uninfected tracer cats, there was no evidence of horizontal transmission of FeLV-myc viruses, suggesting that these viruses arise de novo in individual cases of thymic lymphosarcoma. PMID- 3036719 TI - The relationship of collagenolytic activity to stage of human colorectal carcinoma. AB - Collagenolytic enzymes produced by tumor cells are believed to play a significant role in the destruction of surrounding normal tissue and, in certain experimental animal systems, the ability of tumor cells to degrade type-IV collagen (basement membrane collagen) correlates positively with those cells' metastatic capacity. We measured collagenolytic activity levels of extracts from freshly excised colorectal carcinoma tissues and of conditioned media from primary organ culture (total of 114 tissues from 53 patients) by using purified radiolabelled type-I (rat tail) and type-IV (mouse Engelbreth-Holm-Swarm [EHS] sarcoma) collagens. Both type-IV and type-I collagenolytic activity levels of extracts from tumor and adjacent mucosa ranged from less than 1 to 80 ng/hr/mg wet tissue, and no significant differences between mucosa and carcinoma tissues were observed. In conditioned media, the type-IV collagenolytic activity was low for normal mucosa and benign tumors and slightly higher for carcinoma than for mucosa. In 5 of 32 primary tumors, collagenolytic activity levels were 2-5 times higher than in the rest of the tumors and mucosal tissues. There were no significant differences in collagenolytic activity levels of conditioned media and tissue extract from colorectal carcinoma of different Dukes' stages. Deep and superficial areas of primary tumors released similar type-IV collagenolytic activity levels, suggesting that there was little intratumoral heterogeneity in the release of this enzyme. PMID- 3036721 TI - The biological activity of hydrogen peroxide. III. Induction of Epstein-Barr virus via indirect action, as compared with TPA and teleocidin. AB - Hydrogen peroxide (H2O2) was found to be a potent inducer of the production of the early antigen complex (EA) and/or virus capsid antigen (VCA), determined by Epstein-Barr virus (EBV). Maximum synthesis of EBV antigens was induced by 0.2 mM H2O2, 5 days after a single 10-min treatment, in both B95-8 cells (30.2%) and P3HR-I cells (17.4%). These induction frequencies by H2O2 of proteins encoded by the EBV genome were almost the same as those obtained by 10 min treatment with the potent inducers, 12-O-tetradecanoylphorbol-13-acetate (TPA) and teleocidin. In combination with n-butyrate (4mM), H2O2 showed an additive induction of EBV antigens (1.9- and 1.7-fold in B95-8 and P3HR-I cells, respectively) and was as efficient as TPA and teleocidin. H2O2 induced EBV antigens at a very low level (less than 1%) in Raji cells by itself, but it induced EBV antigens synergistically in combination with n-butyrate (about 25-fold). In combined treatments using H2O2, TPA and teleocidin, the combination of H2O2 and TPA or H2O2 and teleocidin showed additive effects on the induction of synthesis of EBV antigens, but the combination of TPA and teleocidin showed almost the same induction level as that produced by H2O2, TPA or teleocidin alone. The inducing activities of H2O2, TPA and teleocidin were suppressed completely, in the case of H2O2, and slightly, in the cases of TPA and teleocidin, by treatment with catalase. Moreover, the effects of H2O2 were largely suppressed by scavengers of hydroxyl radical (X OH) and singlet oxygen (1O2), but not by superoxide dismutase (SOD), whereas the induction of EBV proteins by TPA and teleocidin was largely suppressed by SOD, but only slightly by scavengers of X OH and 1O2. Thus, the biological actions of H2O2 on the activation of the EBV genome seem to be essentially different from those of TPA and teleocidin, though the biological actions of TPA and teleocidin may be partially ascribed to those of H2O2. PMID- 3036722 TI - Studies on the polyoma tumor-specific transplantation antigen (TSTA): selection and characterization of TSTA-negative segregants from somatic hybrids. AB - Two polyoma-TSTA-negative variants were selected independently from a polyoma fibrosarcoma/Moloney lymphoma somatic hybrid, by repeated passages in polyoma virus-preimmunized mice. One of the variants had lost all its polyoma DNA, while the other only retained a deleted piece of its integrated polyoma DNA. In contrast to the parental hybrid clone, none of the variants produced detectable amounts of T-antigens. This finding indicates that a detectable expression of the products of the polyoma virus early genome, the T-antigens, is important either directly or indirectly for the expression of TSTA. PMID- 3036723 TI - Characterization of neural cell adhesion molecules (NCAM) expressed by Ewing and neuroblastoma cell lines. AB - The status of the neural cell adhesion molecule NCAM gene which is mapped to human chromosome 11q23-24 has been investigated in Ewing-tumor-derived cell lines which present the t(11;22)(q23-24;q12) translocation characteristic of this malignancy. No rearrangement was detected when 2 different non-overlapping probes to mouse NCAM were used. The expression of the NCAM gene was analysed at both the protein and messenger levels in material extracted from Ewing cell lines, human neuroblastoma cell line and fetal mouse brain. Immune blot and immunoprecipitation studies showed that the neuroblastoma cell line contained more NCAM material than the Ewing lines. In neuroblastoma but not in Ewing, the NCAM material had the electrophoretic characteristics of molecules with long polysialic acid chains. After treatment with endosialidase, the diffusely migrating neuroblastoma material was resolved into 3 discrete bands of 120, 140 and 180 kDa. In Ewing extract, high-molecular-weight NCAM species were also detected with a 3-band pattern more reminiscent of mature brain. Endoglycosidase F treatment of Ewing NCAM indicated that all 3 species were largely N glycosylated. Northern blot analysis confirmed that NCAM was expressed more abundantly in neuroblastoma than in Ewing cell lines. Among the 4 NCAM messengers (7.0, 6.5, 4.3 and 4.1 kb) detected in the neuroblastoma, the 6.5 kb species was largely predominant. The Ewing messenger RNA pattern was clearly different as the largest 7.0-kb species was virtually absent and the other bands were of similar intensities. PMID- 3036724 TI - Multimodality treatment with surgery in small-cell lung cancer. AB - The paper reviews the role of surgery in the local treatment of the primary tumour of small-cell bronchial carcinomas (SCLC) and the preliminary results of a multicentre cooperative prospective randomized trial with multimodality treatments of SCLC. This randomized cooperative trial was activated in 1979 comparing a new sequential intermittent polychemotherapy (sq CT) of three different drug combinations alternatingly given intermittently within one year after surgery, with a standard chemotherapy (CT) of one combination of four drugs given intermittently within three years after surgery. Observations on 25 patients at nine different cooperating surgical departments showed that sq CT was applicable and unexpected side-effects did not occur. Live table curves, calculated from 56 patients up to February 1986, indicate the improvement of the survival rate five years after surgery for the cure of 25 patients receiving sq CT after surgery in comparison with 31 patients receiving standard chemotherapy (36% : 22%). Following discussions of this and other results at the symposium "Adjuvant Therapy of Small-Cell Bronchial Carcinoma" at the 13th Congress of the International Society of Chemotherapy (ISC) 1983 in Vienna, a related new protocol was designed and activated by a further enlarged ISC-Study-Group in October 1984. This protocol for a two-armed randomized cooperative trial for patients operated for T1, 2N0M0SCLC is comparing a chemotherapy-combination 1 (CAV)--as also used in a trial of the North American Lung Cancer Study Group-0- with chemotherapy 2 (sq CT) as used in our forementioned previous study. PMID- 3036725 TI - Radioprotective activity of ethylcellulose microspheres containing WR 2721, after oral administration. AB - Ethylcellulose microspheres containing WR 2721 were prepared by the emulsion solvent evaporation technique. No significant loss or degradation of this phosphorothioate was noted during preparation. Oral administration of these microspheres to mice gave an important lowering of WR 2721 toxicity and an enhancement of its radioprotective activity with a D.R.F. of about 1.7-1.8 over 2 3 h. This action is explained by the protection of WR 2721 from acid hydrolysis and degradation in the gastro-intestinal tract. The adsorption of a fraction of WR 2721 onto the surface of microspheres constitutes an inconvenience. This study confirms the interest of such carriers for providing important sustained radioprotection after oral administration. PMID- 3036727 TI - Alzheimer's disease and the pivotal role of the hypothalamus and the intrinsic opioid system. AB - Although the "cholinergic hypothesis" of the pathophysiology of Alzheimer's disease has received great attention during the past years, it has recently come under increasing criticism specifically owing to failure of therapeutic endeavors based on this premise. As the potential broad role of the intrinsic opioids in the neurochemical modulation of diverse brain functions emerges, the realization that these data may be reconciled to a unifying hypothesis underlying the nature of some chronic dementing diseases (including Alzheimer's disease, Korsakoff disease and Parkinson's disease) occurs. Certain specific characteristics of the known pathologic changes and neurotransmitter deficits of Alzheimer's disease may be explained based on an early vulnerability of the hypothalamus combined with derangements of endorphinergic functions which follow. The latter may be implicated in the subsequent degeneration of structures receiving projections from the arcuate nucleus of the hypothalamus. This is based on the known role of endorphins in the modulation of central neurotransmitters and specifically acetylcholine activity. In addition, the reciprocal neuroendocrine and neuroimmunologic interactions mediated through the hypothalamus, may be of further importance in the evolution of Alzheimer's disease. PMID- 3036726 TI - Radiation-induced DNA single-strand breaks in the intestinal mucosal cells of mice treated with the radioprotectors WR-2721 or 16-16 dimethyl prostaglandin E2. AB - Both S-2-(3-aminopropylamino) ethylphosphorothioic acid (WR-2721) and 16-16 dimethyl prostaglandin E2 (dm PGE2) protected the intestinal clonogenic cells to some degree from the effects of 137Cs gamma-irradiation. The D0 was increased from 1.1 +/- 0.12 Gy in controls to 1.55 +/- 0.48 Gy in 16-16 dm PGE2 treated and 2.12 +/- 0.20 Gy in WR-2721 treated mice. Both agents also increased the shoulder of the clonogenic-cell survival curve. Studies were done to measure the effects of these two different radioprotectors on radiation-induction of DNA single strand breaks in cells comprising the murine intestinal mucosa. The number of DNA single-strand breaks increased with increasing doses of gamma-rays in animals killed immediately following exposure. WR-2721 reduced the number of initial radiation-induced DNA single-strand breaks when given one-half hour before exposure; the time of maximum protection. In contrast, 16-16 dm PGE2 given 1 hour before irradiation (the time required to afford maximum protection from radiation cytotoxicity) did not reduce the number of initial DNA breaks. Both agents impeded the rate of rejoining of DNA breaks with increasing time after irradiation. However, the relationship between these effects on the rate of strand rejoining and cell survival is unknown. These results suggest that either both agents are similarly distributed within the cells but the mechanisms of radioprotection are different, or the mechanisms by which these agents protect are similar, but the two agents affect different subcellular targets, the protection of which contributes to increased cell survival. PMID- 3036728 TI - Diethylstilbestrol counteracts barbiturate narcosis and hypothermia in male mice. AB - Treatment with DES one hour prior to pentobarbital injection resulted in diminution of the narcotic sleep and hypothermia usually found after pentobarbital (50 mg/kg 1) injection in male mice. The effect was biphasic: significantly countered relative to saline pretreated controls at low (.001-.10 mg/kg 1) and at high (10-50 mg/kg -1) DES doses only. Giving DES alone did not change core body temperature compared to saline injected controls, at either 25 degrees C or at 33 degrees C. At 33 degrees C, neither PeB narcosis nor body temperature loss was significantly inhibited by DES. Possible cytoplasmic, nonsex differentiated bases for these estrogen effects on barbiturate action in the brain is discussed. PMID- 3036730 TI - Opioid receptor differentiation and Gilles de la Tourette syndrome. PMID- 3036731 TI - Adrenocorticotrophic hormone (ACTH) and movement disorders. PMID- 3036732 TI - The hypothalamus in Parkinson's disease. PMID- 3036729 TI - Spinal dopamine mechanisms and primary sensory symptoms in Parkinson's disease. AB - Primary sensory symptoms occur in approximately 40% of patients with Parkinson's disease. Although the pathophysiology of these symptoms is poorly understood, the recent discovery of diencephalospinal dopaminergic neuronal pathways potentially implicates this system in the etiology of these symptoms. The possible contribution of the hypothalamus with its connections to descending midbrain structures and the spinal cord to the etiology of primary sensory symptoms in Parkinson's disease are discussed. PMID- 3036733 TI - Mechanisms of vascular actions of PAF. AB - The vascular actions of platelet-activating factor (PAF) in the coronary circulation in vitro (rat, guinea-pig) and the intracranial circulation in vivo in the anaesthetized pig have been examined. PAF elicited platelet-independent coronary vasoconstrictor responses which were associated with the release of vasoactive amounts of cyclo-oxygenase and lipoxygenase products. Vasoconstrictor responses in rat hearts showed a greater dependence on the release of leukotriene (LT) C4 than those in guinea-pig hearts. In addition, the latter species was considerably more sensitive to the coronary vasoconstrictor actions of PAF. Intra arterial administration of PAF in the pig evoked biphasic changes in intracranial blood flow comprising a transient increase followed by prolonged decrease. Indomethacin treatment inhibited the decreases in blood flow whereas the transient increases in flow became more pronounced. These results suggest that PAF acts, as least partly, via the release of vasoactive arachidonic acid metabolites. The potency and duration of PAF-induced vasoconstriction suggest a role for this pro-inflammatory phospholipid in the reduction of cerebral and coronary blood flow in pathological conditions. PMID- 3036734 TI - Ebselen: a new approach to the inhibition of peroxide-dependent inflammation. AB - Ebselen is a novel organo-selenium compound which catalytically inactivates peroxides in vitro in a manner similar to that of glutathione peroxidase (GSH Px). In addition, ebselen also inactivates leukotriene B4 (LTB4) generated by pig leukocytes in vitro by isomerizing this eicosanoid to its biologically inactive 6 trans isomer. In vivo, ebselen is a weak oral inhibitor of carrageenan paw oedema and adjuvant arthritis in the rat, differentiating it from classical NSAIDs such as indomethacin and diclofenac. In contrast, oral ebselen, like (intra-articular) catalase, is an effective inhibitor of monoarthritis induced in mice with amidated glucose oxidase (aGO) and dose-dependently inhibits cobra-venom-factor induced paw oedema in rats. Indomethacin and piroxicam are weakly active or ineffective in these models. The data indicate that ebselen is likely to be useful in the therapy of inflammatory conditions in which reactive oxygen species, such as peroxides, play an aetiological role. PMID- 3036735 TI - Differential susceptibility of human mononuclear cells to infection with HTLV-I. AB - Infection with human T-cell leukaemia/lymphoma (HTLV-I) preferentially affects T cells of the OKT-4 phenotype. The aim of the present study was to determine whether distinct T-cell subsets exhibit differences in susceptibility to virus infection. T cells from peripheral blood were separated according to cell densities by 7-step Percoll gradients. Separated T-cell subpopulations were infected with HTLV-I, using cocultivation with irradiated virus producer MT-2 cell line. Percentages of HTLV-I-infected cells and their phenotypes were assayed by immunofluorescence assay (IFA), using highly specific mouse monoclonal antibody directed against HTLV-I P-19 core protein, and other surface markers. The results showed that different T-cell subpopulations were susceptible to HTLV I infection with the exception of large granular lymphocytes (LGL) which exhibit high cell-mediated natural cytotoxicity (CMNC). PMID- 3036736 TI - Viral-bacterial pneumonia in calves: effects on plasma eicosanoids and long chain fatty acids. AB - In the pathogenesis of bovine pneumonic pasteurellosis, immunodepression induced by stress or respiratory viral infection permits superinfection of the lungs with Pasteurella hemolytica, which results in exudative fibrinous pneumonia. Therefore, bovine pneumonic pasteurellosis was induced by sequential inoculations of calves with bovine herpes virus-1 (BHV-1, 3 X 10(7) tissue culture infectious dose 50 (TCID50)/nostril), followed 3 days later by challenge with P. hemolytica (15 X 10(9) colony-forming units (cfu) intratracheally). To study the pathogenic mechanisms of the disease, we examined the alterations in plasma prostaglandins (PG), thromboxane B2 (TxB2), histamine, serotonin and long-chain fatty acids (LCFA) during BHV-1 infection alone and after challenge exposure to P. hemolytica (i.e. during BHV-1-pneumonic pasteurellosis). BHV-1 infection alone markedly increased plasma PGE but modestly elevated PGF2 alpha, TxB2 and arachidonic, oleic and palmitic acids. After challenge with P. hemolytica, the levels of plasma arachidonic, oleic, and palmitic acids, together with PGE and 6-keto-PGF1 alpha, were elevated markedly, in association with clinical signs of bovine pneumonic pasteurellosis. However, PGF2 alpha and stearic acid increased only transiently whereas TxB2 was unchanged from the control. On the other hand, plasma linoleic acid, histamine and serotonin remained unaltered. These results indicate enhanced eicosanoid biosynthesis and disproportionate rises in LCFA during BHV-1 pneumonic pasteurellosis. While LCFA are needed for energy metabolism, eicosanoids may mediate the immunologic, inflammatory and pulmonary vascular reactions leading to the clinico-pathologic features of BHV-1 pneumonic pasteurellosis. PMID- 3036737 TI - Mitochondrial oxidative capacity in gestosic pregnancy. PMID- 3036739 TI - Characterization of a murine tumor of spontaneous origin with selective hepatic metastasis. AB - The primary objective of this study was to establish the identity and characteristics of a murine uterine tumor of spontaneous origin in a C57Bl/6 Ros mouse that selectively metastasized to the liver. The primary tumor contained a minimum of two different cell types. One cell type was elongated and spindly and readily adhered to a plastic coated surface. From it, a permanent cell line, termed RCS-1 was established. The RCS-1 cell line was poorly tumorigenic in normal C57Bl/6 Ros mice. The other cell type, designated RCS-2, was more rounded in structure. It was maintained in vivo, enzymatically dissociated and used after short-term in vitro cell culture. The RCS-2 cells had phagocytic vacuoles, Fc, and C3 receptors. These cells phagocytosed antibody coated SRBCs and opsonized zymosan. Furthermore, these cells generated the superoxide anion. Thus, the RCS-2 cells are of macrophage origin. The spontaneous and experimental pattern of metastasis of the RCS-2 tumor cells was established. The RCS-2 tumor cells selectively metastasized to the liver by the hematogenous route. Metastatic RCS-2 tumor cells obtained from the liver (RCS-2M) retained their macrophage characteristics. These studies indicate that this murine uterine tumor of spontaneous origin consists of two cell types. One cell type is a reticulum cell sarcoma. PMID- 3036738 TI - Metastatic heterogeneity in a spontaneously metastatic HSV-2 induced hamster fibrosarcoma: association of phenotypic and genotypic properties with metastatic potential. AB - A spontaneously metastatic tumour of hamsters and cell lines derived from its in vivo metastatic deposits, or from in vitro cloning were compared for immunobiological and genotypic variation and correlation sought with regard to metastatic potential. Cell lines were established from seven individual lung metastases following primary tumour resection of the parent (HSV-2-333-2-26) cell line. When inoculated subcutaneously, and upon resection of the subsequent tumour mass, four cell lines (MetA, MetB, MetE and MetF) demonstrated greater, one similar (MetG) and two (MetC and MetD) less metastatic capacity compared with the parent tumour line. In further studies cell lines were established in vitro by single cell cloning of the parent tumour either by limiting dilution or soft agar cloning. In vivo tumour resection experiments showed eight cell lines (S4, S7, S4A, S7A, S7B, C1.1, C1.4 and C1.5) to have an increased metastatic potential and five cell lines (S8, S9, S8B, S9D and S9E) a decreased metastatic potential compared to the parental line. Karyotypic analysis of the cells revealed that all highly metastatic cell lines were of a near diploid genotype, whilst non- and weakly metastatic cell lines, including the parental line, were aneuploid or near tetraploid. The immunobiological characteristics of these cell lines was studied. Assessment of in vivo immunogenicity showed that eight clones (MetA, MetB, MetE, MetF, S7A, C1.1, C1.4 and C1.5) were non-immunogenic whilst the parental tumour line and three clones (MetC, MetD and MetG) exhibited a strong transplantation rejection antigen; immunogenicity showed an inverse correlation with metastatic potential. Susceptibility to NK cytolysis was demonstrated for cell lines exhibiting a weak or non-metastatic/immunogenic phenotype. The origin of metastatic variants and their association with genotype and immunobiological properties is discussed. PMID- 3036740 TI - Tissue factor-dependent activation of platelets by cells and microvesicles of SK OS-10 human osteogenic sarcoma cell line. AB - Cultured SK-OS-10 cells (human osteogenic sarcoma metastatic to lung) shed microvesicles (dia. 300-1000 nm) that contained procoagulant and proaggregatory activities inhibitable by hirudin, by anti-tissue factor antibody and by phospholipase A2. These results show that SK-OS-10 cells belong to a group including U87MG human glioblastoma and HL-60 promyelocytic leukemia in which these activities are due to a thrombin-dependent mechanism arising from the presence of tissue factor on the surface of the tumor cells and their shed microvesicles. PMID- 3036742 TI - Effects of cyclosporine A on clinical and immunological parameters in herpes simplex keratitis. AB - The immunosuppressive effects of cyclosporine A (CyA) on the clinical and antiviral immune responses were examined in experimental herpes simplex virus (HSV) keratitis in the rabbit in order to clarify the role that immune lymphocytes play in herpetic stromal disease. Cyclosporine A was administered intramuscularly to rabbits daily starting from the time of corneal infection with HSV until day 14 postinfection. Control HSV-infected rabbits received daily injections of the solvent vehicle alone. HSV-infected rabbits receiving CyA treatment showed more severe and persistent stromal keratitis, and a greater incidence and duration of virus recovery from the cornea. Suppression of cellular immune responses to T cell mitogens, B cell mitogens (anti-rabbit immunoglobulins), and HSV antigens were observed in the CyA treatment group. These results show that in CyA-treated HSV-infected rabbits the antiviral immune responses are inhibited. Acute viral infections with cytopathic viruses such as HSV may therefore be more dramatic, suggesting that CyA may facilitate the potentiation of HSV infections ordinarily suppressed by immune cells. PMID- 3036743 TI - Elderly man with hematuria and a pelvic mass. Mucinous adenocarcinoma of the urachus. PMID- 3036741 TI - Origin of urea-soluble protein in the selenite cataract. Role of beta-crystallin proteolysis and calpain II. AB - Nuclear cataract resulting from an overdose of selenite was characterized by a five-fold increase in nuclear urea-soluble protein. The origin of this urea soluble protein was examined by two-dimensional electrophoresis, immunoblotting with monospecific antisera against rat lens crystallins, and tryptic mapping. Cataractous urea-soluble protein was primarily composed of insolubilized beta- and gamma-crystallin polypeptides. Polypeptides from cataractous urea-soluble protein, and normal beta L-crystallin aggregates were compared by tryptic mapping. Approximately 19% of the urea-soluble protein from opaque nuclei was composed of 24.7 and 24.0 K polypeptides derived by limited proteolysis of 26.5 K beta L-crystallin polypeptide. Incubation of 26.5 K beta-crystallin polypeptide with purified rat lens calpain II in vitro caused production of fragments with similar molecular weights to polypeptides found in cataractous lenses. These results support the hypothesis that proteolysis may contribute to formation of urea-soluble protein in selenite cataract. PMID- 3036745 TI - Myasthenia-like syndrome induced by cardiovascular agents. Report of a case. AB - The case of a myasthenia-like syndrome induced by cardiovascular drugs is reported. The clinical and electrophysiological features of the case are discussed. PMID- 3036744 TI - Quantitative determination of acylphosphatase levels in horse tissues by enzyme linked immunosorbent assay. AB - A non competitive enzyme-linked immunosorbent assay (ELISA) specific for horse muscle acylphosphatase (E.C. 3.6.1.7.) has been developed. The purified anti acylphosphatase antibodies were immobilized by passive absorption to a solid phase support and incubated with known and unknown amounts of antigen. The antibody-acylphosphatase complex was quantified using the same antibody conjugated to horseradish peroxidase. The assay yields positive reactions with as little as 0.05 ng of antigen, with intra- and interassay coefficients of variation of 5% and 7%, respectively. On the basis of this assay we developed a more sensitive test than the optical one, using benzoyl-phosphate as substrate, for acylphosphatase determination. By means of this test, the presence of the enzyme in horse tissue homogenates was evaluated under conditions in which the optical test failed to distinguish the acylphosphatase activity from that of other enzymes. PMID- 3036746 TI - Polyneuropathy and systemic vasculitis. An electrophysiological study. AB - A review of cases of systemic vasculitis followed for one year in a rheumatology department resulted in the detection of 18 patients who on the clinical and electrophysiological evidence had symmetrical distal polyneuropathy. A moderate impairment of the motor conduction velocity was present in 7 patients, with electromyographic neurogenic changes in the distal lower limbs of all but one patient. The sensory action potential of the sural nerve was bilaterally absent in one case, and its amplitude was reduced in 14 out of 16 patients, with a decreased sensory conduction velocity in 9 cases. The sural nerve biopsy, performed in 6 patients, was prevalently suggestive of previous axonal degeneration. This investigation illustrates the spectrum of diffuse peripheral nerve involvement associated with systemic vasculitis. A variable impairment of the sensory action potential, ranging from slight decrease of amplitude to no response, is the most common finding. Conduction velocities appear to be relatively spared. PMID- 3036747 TI - Hypothesis: chronic benign daily headache is an immune disorder with a viral trigger. PMID- 3036748 TI - Sustained headache syndrome associated with tender occipital nerve zones. PMID- 3036749 TI - Identification of blood leukotrienes in classical migraine. PMID- 3036750 TI - Some measurements of the equilibrium factor for 222Rn daughters in houses. PMID- 3036751 TI - Silylation studies of 6-APA by 1H NMR. PMID- 3036752 TI - The in vivo effects of tetrahydrocannabinol on the activity of lactate and succinate dehydrogenases in rat preovulatory follicles. AB - The effects of delta 9-tetrahydrocannabinol on the activities of lactate and succinate dehydrogenases in the theca interna and membrana granulosa of rat preovulatory follicles have been analysed microdensitometrically using the same injection regime employed in a previous study on steroidogenic enzymes. A small but statistically significant (18%) decrease in succinate dehydrogenase activity was observed in the theca interna, but none in any region of the membrana granulosa. Lactate dehydrogenase activity was unaffected by THC administration. Thus, a dosage and regimen sufficient to cause significant decreases in the activities of steroidogenic enzymes had little effect on succinate and lactate dehydrogenases in rat preovulatory follicles. PMID- 3036753 TI - Improved local control of thoracic disease in small cell lung cancer with higher dose thoracic irradiation and cyclic chemotherapy. AB - Over the past decade, improvement in survival has developed for patients with small cell lung carcinoma (SCLC) due to treatment strategies that include: cyclic combination chemotherapy, thoracic irradiation, and prophylactic cranial irradiation. In this study, we assess the outcome of treatment with initial cyclic combination chemotherapy including: cyclophosphamide, VP 16-123 and methotrexate combined with radiotherapy (RT), 6000 cGY [corrected] to the thorax for patients with limited disease and 3000 cGy [corrected] for patients with extensive disease. Forty-six patients are evaluated: 26 patients with limited disease and 20 with extensive disease. In patients who received 6000 cGy [corrected], to thoracic lesions, in combination with chemotherapy, administered for 3 courses prior to and following RT, the rate of clinically detected failure in the thorax was 3.8%. Morbidity was considered acceptable, although the occurrence of encephalopathy in 6 of 19 cases who received cranial irradiation, 3000 cGy [corrected], and concomitant chemotherapy was a serious consequence. Control of the primary tumor achieved by the use of higher dose RT is shown to be superior to that observed at lower doses of RT. This suggests that for the small cohort of patients whose disease is truly limited at the time of diagnosis, therapeutic regimens, which include higher dose RT, could increase the number of long term survivors of SCLC. PMID- 3036754 TI - Comparison of populations of human faecal bacteria before and after in vitro incubation with plant cell wall substrates. AB - Human faecal slurries were incubated anaerobically with larchwood xylan, oat spelt xylan, wheat bran, apple cell walls or sugar beet pulp as sole carbon sources. The populations which developed during incubation were different from the inoculum, the most marked changes being an increase in the number of Bacteroides species and a decrease in the number of Fusobacterium species for all carbon sources tested. With a water-soluble preparation of larchwood xylan the population was dominated by species able to ferment this substrate, in contrast to the population which developed with the insoluble substrates. The ability to use one plant cell wall substrate appeared to be related to the ability to use others. Strains capable of using plant cell wall substrates included Bacteroides spp., Clostridium clostridiiforme, Bifidobacterium longum, Fusobacterium spp. and Escherichia coli. When incubation with two contrasting substrates (bran and larchwood xylan) was replicated, the populations which developed were reproducibly different from the inoculum and from each other. PMID- 3036755 TI - Some properties of different skeletal muscle fiber types: comparison of reference bases. AB - Several biochemical components of the white portion of the gastrocnemius (WGM), plantaris (PM), and soleus (SM) muscles of the rat and middle gluteal (MGM) muscle of the horse were compared based on wet and dry weight, protein, and total creatine concentrations ([TCr]). The water content was similar for the rat hindlimb muscles, however, the concentrations of protein, ATP, phosphocreatine (PCr), creatine, and glycogen ranked as SM less than PM less than WGM for all reference bases except total creatine. In contrast, concentrations of ATP, creatine, and PCr were similar in all muscles studied when expressed as ratios of [TCr]. Horse MGM had the lowest percent of water and protein per gram wet or dry weight but highest glycogen concentration of the muscles studied, irrespective of the reference base used to express concentrations. Coefficients of variation were lowest when muscle constituents were related to [TCr]. It is concluded that expressing muscle constituents relative to total creatine results in the smallest variation and is a good method for making comparisons between muscles of similar fiber composition. However, essential information concerning different types of muscle may be lost when this reference base is used. PMID- 3036756 TI - Exercise-induced functional desensitization of canine cardiac beta-adrenergic receptors. AB - To test the hypothesis that the high levels of endogenous catecholamines associated with strenuous exercise produce functional desensitization of cardiac beta-adrenergic receptors, we measured the bolus chronotropic dose of isoproterenol necessary to produce a 25-beats/min increase in heart rate (CD25) in the resting state and after the return of heart rate to resting levels after 60 min of treadmill running in 13 normal dogs. Immediately after exercise, 12 of 13 dogs were less sensitive to the chronotropic effects of beta-adrenergic receptor stimulation: mean CD25 increased from 1.16 +/- 0.17 to 3.50 +/- 0.98 micrograms (P less than 0.02). A similar reduction in isoproterenol sensitivity was evident regardless of whether testing was performed in the presence or absence of vagal blockade with atropine. By 3 h after exercise, CD25 had returned to the preexercise level, with no further change noted 24 h after exercise. There was no change in the CD25 when measured serially in three unexercised dogs. We conclude that a single bout of dynamic exercise is sufficient to produce a significantly decreased chronotropic responsiveness to isoproterenol. This phenomenon may represent an acute but transient desensitization of cardiac beta adrenergic receptors. PMID- 3036757 TI - Reproducibility of the multiple inert gas elimination technique. AB - Although measurement errors in the multiple inert gas elimination technique have a coefficient of variation of approximately 3%, small biological fluctuations in ventilation, blood flow, or other variables must contribute additional variance to this method of assessing ventilation-perfusion (VA/Q) mismatch. To determine overall variance of computed indices of VA/Q mismatch, an analysis of variance was carried out using a total of 400 duplicate pairs of inert gas samples obtained from canine (N = 118) and human (N = 282) studies in the past 2 years. In both sets VA/Q mismatch ranged from minimal (2nd moment of ventilation and blood flow distributions, log SDV and log SDQ, respectively approximately equal to 0.3 each) to severe (log SDV and log SDQ approximately equal to 2.0). Differences between duplicate log SD values were computed and found to be a constant fraction of the mean log SD of each duplicate pair, averaging 13% for both canine and human ventilation and blood flow data. The resultant coefficient of variation for a single measurement of log SD about its mean averaged 8.6% for all data combined. This analysis demonstrates excellent reproducibility of these dispersion indices over a wide range of conditions, and if the mean of duplicate values is used, thus reducing variability by square root 2 to 6.1%, log SD can be estimated with an approximately 95% confidence limit of +/- 12%. PMID- 3036758 TI - Concentration-dependent interaction of theophylline with d-tubocurarine. AB - The interaction of theophylline with d-tubocurarine chloride (dTC) was examined in rabbits. After steady-state subtherapeutic (less than 10 mg/l), therapeutic (10-20 mg/l), and toxic (greater than 20 mg/l) concentrations of theophylline, dose-response curves for dTC were determined and compared with controls that received no theophylline. At therapeutic concentrations of theophylline the effective dose for 50% inhibition of twitch (ED50) for dTC (mean +/- SE, 0.115 +/ 0.016 mg/kg) was significantly shifted to the left in comparison with the control (0.165 +/-0.008 mg/kg). The ED50 of dTC for the subtherapeutic group was 0.143 +/- 0.011 mg/kg, which was less than the control but not of statistical significance (P = 0.1). The ED50 for the toxic theophylline group was 0.168 +/- 0.003 mg/kg, which was not significantly different from controls but significantly different from the theophylline therapeutic and subtherapeutic groups. Thus, toxic concentrations of theophylline reversed the potentiating effects of therapeutic and subtherapeutic concentrations of dTC dose-response curves. Therefore, depending on concentration, theophylline exhibits a biphasic interaction with dTC. Surgical patients on theophylline may require less dTC intraoperatively. More importantly, the use of theophylline in the postoperative period to reverse anesthetic effects may result in recurarization. PMID- 3036760 TI - Effect of flow and surface area on angiotensin-converting enzyme activity in rabbit lungs. AB - Pulmonary angtiotensin-converting enzyme (ACE) is located on the luminal surface of pulmonary microvasculature. Multiple indicator-dilution techniques have been used to measure pulmonary ACE activity in vivo and in isolated lungs. These studies suggest that ACE activity is depressed in several forms of acute lung injury. Depression of ACE activity may reflect impaired substrate delivery to enzyme sites because of flow-related reduction of perfused surface area. To assess the role of altered microvascular flow and surface area in the measurement of ACE activity, we utilized similar techniques to estimate the apparent Km and Vmax of pulmonary ACE in isolated, Krebs-perfused rabbit lungs. Km is an estimate of the affinity of a synthetic ACE substrate, [3H]benzoyl-phenyl-alanyl-alanyl proline ([3H]BPAP), for ACE and should not be influenced by the rate of substrate delivery to luminal enzyme sites. Conversely, Vmax is an index of the number of ACE sites and should be influenced by perfusion changes that alter the number of perfused sites (recruitment or derecruitment). When isolated lungs were subjected to physiological maneuvers designed to increase or decrease perfused surface area, apparent Vmax increased or decreased respectively. Apparent Km was not altered by these maneuvers. Km and Vmax were independent of changes in perfusion rate when surface area was held constant. Thus these parameters should be useful in evaluating perfusion changes in normal and injured lungs. PMID- 3036759 TI - Correlation of pulmonary ACE activity and capillary surface area during postnatal development. AB - Although a considerable amount of information is available regarding the remodeling and growth of the pulmonary arterial circulation, relatively little is known regarding postnatal development of the pulmonary microcirculation. We hypothesized that the maximal velocity (Vmax) of pulmonary angiotensin-converting enzyme (ACE) activity, measured from indicator-dilution outflow curves using a synthetic substrate, 3H-labeled benzoyl-phenylalanyl-alanyl-proline (BPAP), is directly related to the capillary endothelial cell surface area in the lungs of developing lambs. Accordingly we measured apparent kinetics of pulmonary ACE activity in 22 anesthetized ventilated lambs (2-171 days old) and compared our functional assessment to simultaneous in vivo determinations of CO diffusing capacity (DLCO) and postmortem structural assessment of alveolar septal dimensions using stereology and electron microscopy. There was a progressive increase in Vmax of ACE in this age group, with little change in apparent affinity for BPAP. Similar functional manifestation of growth was noted by an age dependent increase in DLCO. Neither Vmax nor DLCO was significantly affected by an increase in left atrial pressure to 19 Torr (via inflation of a balloon in the left atrium), suggesting little recruitment of vessels under conditions of the present protocol. A close correlation was observed when either Vmax for ACE activity or DLCO was plotted vs. capillary endothelial cell surface area. Double logarithmic transformation of capillary endothelial cell surface area, Vmax-ACE and DLCO vs. lung volume revealed power functions with slopes all greater than that predicted from isotropic growth, suggesting selective differential postnatal development of the endothelium of the alveolar septum in lambs from 2-171 days of age. PMID- 3036761 TI - Establishment of an adult rat fibroblast cell line for studies of collagenase regulation. AB - Several dermal fibroblast lines have been established from explants taken from adult rats. The cells have been cultured for 2 yr and possess stable and well defined growth characteristics through subculture 18. The cells are readily stored in liquid nitrogen with good viability after thawing. Collagenase activity secreted into the culture medium of the cells at different periods of growth has been examined. There is an 88% drop in total enzyme activity present in the medium between 4 and 14 d of culture, when the cells were plated to reach confluence at Day 8 to 10. A more pronounced fall is noted at earlier times when the cells are plated at a higher density. The correlation between DNA content of the cell monolayer and enzyme activity was -0.895, indicating a possible relationship between the growth of cells and collagenase release. PMID- 3036762 TI - Purification of Black Moor goldfish melanophores and responses to epinephrine. AB - Two key modifications of the previously reported method for isolation of goldfish xanthophores allowed the isolation and establishment of primary cultures of terminally differentiated melanophores from the Black Moor goldfish (Carassius auratus). First, pretreatment with 10(-4) M epinephrine causing aggregation of the melanosomes and collapse of the dendrites, prevents damage to the melanophores during tissue dissociation and melanophore isolation. Second, maintenance of these cells in culture was successful only when the culture medium was supplemented with fish serum. The purified melanophores attached, flattened, and were maintained in culture for up to 3 mo. Although the morphology of the cultured melanophores is less dendritic than their in vivo counterparts, the melanophores translocate melanosomes in a normal manner except that they exhibit enhanced sensitivity to epinephrine. This epinephrine-induced pigment aggregation, as well as the redispersion of pigment after the removal of epinephrine, can occur in the presence of ethylene glycol-bis (beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid and absence of Ca2+. PMID- 3036763 TI - Cloning and nucleotide sequence of phoP, the regulatory gene for alkaline phosphatase and phosphodiesterase in Bacillus subtilis. AB - Two DNA fragments which complement the alkaline phosphatase-negative mutation phoP of Bacillus subtilis were cloned from a B. subtilis chromosome with the prophage vector phi CM (a derivative of phi 105). One of the fragments contained the regulatory gene phoR in addition to phoP. Nucleotide sequence analysis of the phoP region revealed that the phoP gene product consists of 241-amino-acid residues and that the sequence of these amino acids is extensively homologous with the sequence of the phoB gene product. This protein is the positive regulator for the phosphate regulon in Escherichia coli. It therefore appears that phoP is a regulatory gene for alkaline phosphatase synthesis in B. subtilis. PMID- 3036764 TI - Analysis, cloning, and high-level expression of 2,4-dichlorophenoxyacetate monooxygenase gene tfdA of Alcaligenes eutrophus JMP134. AB - Plasmid pJP4 of Alcaligenes eutrophus JMP134 contains all genes for the degradation of 2,4-dichlorophenoxyacetic acid (2,4-D). Five of these genes, tfdB, tfdC, tfdD, tfdE, and tfdF, have recently been localized and cloned (R. H. Don, A. J. Weightman, H.-J. Knackmuss, and K. N. Timmis, J. Bacteriol. 161:85-90, 1985). Gene tfdA, which codes for the 2,4-D monooxygenase, has now been found by mutagenesis with transposon Tn5. A 3-kilobase fragment of pJP4 cloned in a broad host-range vector could complement the 2,4-D-negative phenotype of two mutants which lacked 2,4-D monooxygenase activity. The cloned tfdA gene was also transferred to A. eutrophus JMP222, which is a cured derivative of JMP134. The recombinant strain could utilize phenoxyacetic acid as a sole source of carbon and energy. Pseudomonas sp. strain B13, containing the cloned tfdA, was able to degrade phenoxyacetic acid and 4-chlorophenoxyacetic acid. Gene tfdA was subcloned and analyzed by deletions. Expression of 2,4-D monooxygenase in Escherichia coli containing a 1.4-kilobase subfragment was demonstrated by radioisotopic enzyme assay, and a protein of 32,000-dalton molecular mass was detected by labeling experiments. A 2-kilobase subfragment containing tfdA has been sequenced. Sequence analysis revealed an open reading frame of 861 bases which was identified as the coding region of tfdA by insertion mutagenesis. PMID- 3036765 TI - Characterization and expression of a cloned tetracycline resistance determinant from Campylobacter jejuni plasmid pUA466. AB - A tetracycline resistance (Tcr) determinant previously cloned from the Campylobacter jejuni plasmid pUA466 (D. E. Taylor, J. Bacteriol. 165:1037-1039, 1986) was localized by restriction endonuclease mapping, subcloning, and Tn1000 insertion mutagenesis to a 2-kilobase region consisting of 1.8- and 0.2-kilobase HincII fragments. Tcr encoded by the cloned fragment (pUOA1) was expressed constitutively in Escherichia coli. A protein with an apparent molecular weight of 68,000 encoded by pUOA1 was produced in an in vitro transcription-translation system and in minicells. Tn1000 insertions which resulted in inactivation of Tcr expression also resulted in an alteration in the 68,000-molecular-weight protein. Some mutants specified a truncated protein, whereas others completely lost the ability to specify the protein. The protein which appears to be involved in the expression of Tcr specified by C. jejuni plasmids is of approximately the same molecular weight as the protein specified by the streptococcal class M determinant. This finding is consistent with our previous results which indicate that homology exists between the Tcr determinant from C. jejuni and a 5-kilobase HincII probe derived from the streptococcal class M determinant. PMID- 3036766 TI - Physical and genetic characterization of the glucitol operon in Escherichia coli. AB - The glucitol (gut) operon has been identified in the colony bank of Clark and Carbon (A. Sancar and W. D. Rupp, Proc. Natl. Acad. Sci. USA 76:3144-3148, 1979). We subcloned the gut operon by using paCYC184, pACYC177, and pBR322. The operon, which is encoded in a 3.3-kilobase nucleotide fragment, consists of the gutC, gutA, gutB, and gutD genes. The repressor of the gut operon seemed to be encoded in the region downstream from the operon. The gene products of the gut operon were identified by using maxicells. The apparent molecular weights of the glucitol-specific enzyme II (product of the gutA gene), enzyme III (product of the gutB gene), and glucitol-6-phosphate dehydrogenase (product of the gutD gene) were about 46,000, 13,500, and 27,000, respectively, as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. PMID- 3036767 TI - Energetics of the initial phase of adhesion of Streptococcus sanguis to hydroxylapatite. AB - The initial adhesion of Streptococcus sanguis 10556 to artificial salivary pellicle and to bare hydroxylapatite was studied at several temperatures between 18 and 37 degrees C. When the natural logarithms of rate constants for adsorption and desorption were plotted against reciprocal temperatures in Arrhenius plots, curved lines were obtained, indicating that the thermodynamic quantities of enthalpy and entropy of activation were temperature dependent. For the bare hydroxylapatite system, the heat capacity (delta Cp = dH/dT) was large and negative. delta Cp was also negative for adhesion to saliva-coated hydroxylapatite, although its value was lower. Negative heat capacities, when coupled with favorable entropy, are often indicative of either electrostatic or hydrophobic interactions. When electrolyte (100 mM ammonium sulfate) was added to the cell-hydroxylapatite bead mixture, the rate and extent of adhesion were decreased. Addition of nonpolar p-dioxane (10% [vol/vol], final concentration) to the mixture enhanced binding. This suggests that electrostatic linkages participate in the primary adhesion of streptococci to both substrata. The strongly positive entropy values and the lesser temperature dependence of the saliva-coated hydroxylapatite system suggest that another entropy-driven process is imposed on the electrostatic linkages. This supports a role for hydrophobicity, suggesting that a combination of electrostatic and hydrophobic forces mediate the initial adhesion of S. sanguis to the salivary pellicle. PMID- 3036768 TI - Bacteriophages of Saccharopolyspora erythraea. AB - Five bacteriophages infecting only Saccharopolyspora erythraea (formerly Streptomyces erythreus) among 43 Streptomyces spp. tested were classified into two groups by phage-host relationships, restriction enzyme mapping, cohesive-end determinations, and Southern hybridizations. phi SE6, the most frequently isolated phage, produced clear plaques on all hosts tested, while phi SE45, phi SE57, phi SE60, and phi SE69 produced turbid plaques. phi SE6 DNA was linear, had a molecular weight of (27.6 +/- 1) X 10(6) and, like the DNAs of phi SE45, phi SE57, and phi SE69, lacked cohesive ends. The characteristic patterns of of ClaI and HindIII restriction digests of phi SE6 DNA and the results of Southern hybridizations with three different ClaI fragments of phi SE6 DNA as probes indicated that phi SE6 DNA was partially circularly permuted and terminally redundant, suggesting that it was packaged by a headful packaging mechanism. Southern hybridization data also showed that phi SE45, phi SE57, and phi SE69 were closely related to phi SE6. phi SE60 DNA, in contrast, had cohesive ends, and restriction mapping plus Southern hybridization data showed that phi SE60 was unrelated to the other four phages. PMID- 3036770 TI - Isolation and characterization of Tn917lac-generated competence mutants of Bacillus subtilis. AB - We isolated 28 mutants of Bacillus subtilis deficient in the development of competence by using the transposon Tn917lacZ as a mutagen. The mutant strains were poorly transformable with plasmid and chromosomal DNAs but were normally transducible and exhibited wild-type resistance to DNA-damaging agents. The mutations were genetically mapped, and the mutants were characterized with respect to their abilities to bind and take up radiolabeled DNA. All were defective in uptake, and some failed to bind significant amounts of DNA. The abilities of the mutant strains to resolve into two buoyant density classes on Renografin gradients were studied. Most resolved normally, but several banded in Renografin only at the buoyant density of noncompetent cells. The genetic mapping studies and the other analyses suggested that the mutations define a minimum of seven distinct com genes. PMID- 3036769 TI - Cloning and nucleotide sequencing of genes for a second type of small, acid soluble spore proteins of Bacillus cereus, Bacillus stearothermophilus, and "Thermoactinomyces thalpophilus". AB - The nucleotide sequences of the single genes coding for the B-type small, acid soluble spore proteins (SASP) of Bacillus cereus, B. stearothermophilus, and "Thermoactinomyces thalpophilus" were determined, and the amino acid sequences of all B-type SASP were compared. While this type of SASP showed significant sequence conservation around the two spore protease cleavage sites, alignment of these sequences required the introduction of gaps, and even then only 19 of the residues were conserved exactly in all five proteins. However, all five B-type SASP did contain a large (27 to 35-residue), rather well-conserved amino acid sequence repeat, and four of the five proteins had well-conserved regions of 14 to 17 amino acids which appeared three times. PMID- 3036772 TI - Facilitated transfer of IncP beta R751 derivatives from the chromosome of Bacteroides uniformis to Escherichia coli recipients by a conjugative Bacteroides tetracycline resistance element. AB - The broad-host-range IncP beta plasmid R751 can mobilize itself from Escherichia coli to Bacteroides spp, but it is not maintained in Bacteroides spp. If R751 carries the Bacteroides transposon Tn4351, it can be integrated into the Bacteroides chromosome. Previously we showed that R751, integrated in the chromosome of Bacteroides uniformis, cannot mobilize itself out of B. uniformis into E. coli or isogenic B. uniformis strains. In this report, we showed that if the Bacteroides conjugative tetracycline resistance element Tcr ERL was coresident with the R751 insertion in B. uniformis, derivatives of R751 were transferred to E. coli, where they were recovered as plasmids. The most common derivatives were R751::Tn4351 and R751::IS4351, but some strains transferred R751 derivatives, containing additional DNA segments ranging in size from 10 to 23 kilobases. These DNA inserts cross-hybridized with chromosomal DNA from B. uniformis which did not carry the Tcr ERL element. Therefore, the inserts appeared to be segments of the wild-type B. uniformis chromosome and were not associated with the Tcr ERL element. The transfer of integrated R751 from B. uniformis was independent of the RecA phenotype of the E. coli recipients and did not appear to be due to transfer of B. uniformis chromosomal DNA, followed by RecA-dependent recombination between homologous IS4351 sequences to form the resultant R751 plasmid derivatives. Consistent with this, no transfer of Tn4351 (associated with the cointegrated R751) from B. uniformis donors to isogenic B. uniformis recipients was detected (< 10(-8)). Our data support the hypothesis that R751 excises from the B. uniformis chromosome by recombination involving flanking Tn4351 or IS4351 sequences and forms nonreplicating circles. The mobilization of these circular forms out of B. uniformis to E.coli is then facilitated by the Tcr ERL element. PMID- 3036771 TI - Oxygen as attractant and repellent in bacterial chemotaxis. AB - Studies of bacterial chemotaxis to oxygen (aerotaxis) over a broad range of oxygen concentrations showed that at high concentrations, oxygen was a repellent of Salmonella typhimurium, Escherichia coli, and some bacilli, whereas it is known that at lower concentrations (less than or equal to 0.25 mM dissolved oxygen), oxygen is an attractant. In a temporal assay of aerotaxis, S. typhimurium in medium equilibrated with air (0.25 mM dissolved oxygen) and then exposed to pure oxygen (1.2 mM) tumbled continuously for approximately 20 s. The oxygen concentration that elicited a half-maximal negative (repellent) response was 1.0 mM for both S. typhimurium and E. coli. The receptor for the negative chemoresponse to high concentrations of oxygen is apparently different from the receptor for the positive chemoresponse to low concentrations of oxygen, since the oxygen concentration that elicits a half-maximal positive (attractant) response in S. typhimurium and E. coli is reported to be 0.7 microM. Adaptation to high concentrations of oxygen, like adaptation to low concentrations of oxygen, was independent of methylation of a transducer protein. Only the response to low oxygen concentrations, however, was altered by interaction with the amidated Tsr transducer in cheB mutants. PMID- 3036773 TI - Cloning and genetic organization of the pca gene cluster from Acinetobacter calcoaceticus. AB - The beta-ketoadipate pathway of Acinetobacter calcoaceticus comprises two parallel metabolic branches. One branch, mediated by six enzymes encoded by the cat genes, converts catechol to succinate and acetyl coenzyme A (acetyl-CoA); the other branch, catalyzed by products of the pca genes, converts protocatechuate to succinate and acetyl-CoA by six metabolic reactions analogous or identical to those of the catechol sequence. We used the expression plasmid pUC18 to construct expression libraries of DNA from an A. calcoaceticus mutant strain from which the cat genes had been deleted. Immunological screening with antiserum to the pcaE gene product, beta-ketoadipate:succinyl-CoA transferase I, resulted in the isolation of a cloned 11-kilobase-pair (kbp) fragment which inducibly expressed all six pca genes under control of the lac promoter on pUC18. The induced Escherichia coli cells formed the six pca gene products at levels 10- to 30-fold higher than found in fully induced A. calcoaceticus cultures, although protocatechuate 3,4-dioxygenase (the iron-containing product of the pcaA gene) from the recombinant strain possessed a relatively low turnover number. An E. coli culture expressing the cloned pca genes quantitatively converted protocatechuate to beta-ketoadipate; failure of the organism to metabolize the latter compound can be most readily ascribed to relatively low pool levels of succinyl-CoA, a required substrate for beta-ketoadipate:succinyl-CoA transferase, in E. coli. The gene order and direction of transcription were determined to be pcACBDFE by identification of enzymes expressed in subclones, by using natural transformation to identify subclones carrying DNA corresponding to dysfunctional alleles in mutant A. calcoaceticus strains, and by restriction mapping of both the 11-kbp fragment and derivatives of the 11-kbp fragment containing Tn5 in the pcaA, pcaB, pcaD, and pcaE genes. The fragment containing the pca gene hybridized strongly and specifically to a previously cloned fragment containing A. calcoaceticus cat genes. PMID- 3036774 TI - Cobalamin (vitamin B12) biosynthetic genes of Salmonella typhimurium. AB - The enteric bacterium Salmonella typhimurium synthesizes cobalamin (vitamin B12) de novo only under anaerobic growth conditions. We initiated a genetic analysis of the cobalamin biosynthetic (cob) gene cluster, Stable cob::lac operon fusions were generated by insertions of a transposition-defective derivative of bacteriophage Mu d1 (Ap lac) into the cob genes. beta-Galactosidase synthesis was repressed in the presence of exogenously supplied cyanocobalamin, demonstrating that the cobalamin biosynthetic pathway was regulated by end-product repression. Transcriptional polarity studies showed that the cob genes responsible for synthesis of the corrinoid intermediate cobinamide (branch I of the pathway) were organized into a single operon. Genes for the synthesis of 5,6 dimethylbenzimidazole (branch II) and the final assembly of the complete cobalamin molecule (branch III) were organized into two or more additional operons. All of the known cob genes (in branches I, II, and III) were transcribed in a counterclockwise direction relative to the S. typhimurium genetic map. These genes are located at 41 map units and near the his operon. No essential genes lie between the his and cob operons. Mutants that carried deletions extending from the his genes into the cob region were isolated and characterized. By using these mutants, a deletion map of the branch I cob operon was constructed and the order of genes (his-cobI-cobIII-cobII) was inferred. PMID- 3036775 TI - Molecular cloning and characterization of a Streptococcus sanguis DNase necessary for repair of DNA damage induced by UV light and methyl methanesulfonate. AB - We developed a method for cloning cellular nucleases from streptococci. Recombinant lambda gt11 bacteriophage containing streptococcal nuclease determinants were identified by the production of pink plaques on toluidine blue O DNase plates. We used this technique to clone a 3.2-kilobase-pair EcoRI fragment with DNase activity from the chromosome of Streptococcus sanguis. The locus was designated don (DNase one) and could be subcloned and stably maintained on plasmid vectors in Escherichia coli. Minicell analyses of various subclones of the don locus allowed us to determine the coding region and size of the Don nuclease in E. coli. The don gene product had an apparent molecular mass of 34 kilodaltons and degraded native DNA most efficiently, with lesser activity against denatured DNA and no detectable activity against RNA. S. sanguis don deletion mutants were constructed by transformation of competent cells with in vitro-prepared plasmid constructs. S. sanguis don deletion mutants retained normal transformation frequencies for exogenously added donor DNA. However, when compared with Don+ wild-type cells, these mutants were hypersensitive to DNA damage induced by UV light and methyl methanesulfonate. An S. sanguis don specific DNA probe detected homology to chromosomal DNA isolated from Streptococcus pneumoniae and Streptococcus mutans Bratthall serogroups d and g. Our results suggested that the don locus was the S. sanguis allele of the previously described S. pneumoniae major exonuclease and was involved in repair of DNA damage. Furthermore, hybridization studies suggested that the don locus was conserved among species of oral streptococci. PMID- 3036776 TI - Alginate biosynthetic enzymes in mucoid and nonmucoid Pseudomonas aeruginosa: overproduction of phosphomannose isomerase, phosphomannomutase, and GDP-mannose pyrophosphorylase by overexpression of the phosphomannose isomerase (pmi) gene. AB - The specific activities of phosphomannose isomerase (PMI), phosphomannomutase (PMM), GDP-mannose pyrophosphorylase (GMP), and GDP-mannose dehydrogenase (GMD) were compared in a mucoid cystic fibrosis isolate of Pseudomonas aeruginosa and in two spontaneous nonmucoid revertants. In both revertants some or all of the alginate biosynthetic enzymes we examined appeared to be repressed, indicating that the loss of the mucoid phenotype may be a result of decreased formation of sugar-nucleotide precursors. The introduction and overexpression of the cloned P. aeruginosa phosphomannose isomerase (pmi) gene in both mucoid and nonmucoid strains led not only to the appearance of PMI levels in cell extracts several times higher than those present in the wild-type mucoid strain, but also in higher PMM and GMP specific activities. In extracts of both strains, however, the specific activity of GMD did not change as a result of pmi overexpression. In contrast, the introduction of the cloned Escherichia coli manA (pmi) gene in P. aeruginosa caused an increase in only PMI and PMM activities, having no effect on the level of GMP. This suggests that an increase in PMI activity alone does not induce high GMP activity in P. aeruginosa. The heterologous overexpression of the P. aeruginosa pmi gene in the E. coli manA mutant CD1 led to the appearance in cell extracts of not only PMI activity but also GMP activity, both of which are normally undetectable in extracts of CD1. We discuss the implications of these results and propose a mechanism by which overexpression of the P. aeruginosa pmi gene can cause an elevation in both PMM and GMP activities. PMID- 3036777 TI - Identification of the promoter of the Bacillus subtilis sdh operon. AB - The Bacillus subtilis sdhCAB operon contains the structural genes for the three subunits of the membrane bound succinate dehydrogenase complex. An sdh-specific transcript of about 3,450 nucleotides was detected in vegetative bacteria. S1 nuclease mapping experiments showed that the sdh operon is transcribed from a sigma-43 promoter; the transcript starts at a guanosine residue 90 base pairs upstream from the first gene of the operon, sdhC. No sdh transcript was found in B. subtilis carrying the sdh-115 mutation, which decreases expression of the sdh operon by more than 99%. The sdh-115 mutation is a G-to-A transition in the -35 region of the sigma-43 promoter. The sdh operon is sensitive to glucose repression. When the sdh promoter region was used to drive transcription of the cat-86 gene this gene also became glucose repressed. PMID- 3036778 TI - Cloning of the cyo locus encoding the cytochrome o terminal oxidase complex of Escherichia coli. AB - The structural genes encoding the cytochrome o terminal oxidase complex (cyo) of Escherichia coli have been subcloned into the multicopy plasmid pBR322 after the Mu-mediated transposition of the gene locus from the bacterial chromosome onto the conjugative R plasmid RP4. Introduction of cyo plasmids into strains (cyo cyd) lacking both terminal oxidases restored the ability of the strains to grow aerobically on nonfermentable substrates. Strains carrying the cyo plasmids produced 5 to 10 times more cytochrome o oxidase than did control strains. The gene products encoded by the cyo plasmids could be immunoprecipitated with monospecific antibodies raised against cytochrome o. The cloned genes will be valuable for studying the structure, function, and regulation of the cytochrome o terminal oxidase complex. PMID- 3036780 TI - Revised genetic map of the distal end of the F transfer operon: implications for DNA helicase I, nicking at oriT, and conjugal DNA transport. AB - The DNA transfer stage of conjugation requires the products of the F sex factor genes traMYDIZ and the cis-acting site oriT. Previous interpretation of genetic and protein analyses suggested that traD, traI, and traZ mapped as contiguous genes at the distal end of the transfer operon and saturated this portion of the F transfer region (which ends with an IS3 element). Using antibodies prepared against the purified TraD and TraI proteins, we analyzed the products encoded by a collection of chimeric plasmids constructed with various segments of traDIZ DNA. We found the traI gene to be located 1 kilobase to the right of the position suggested on previous maps. This creates an unsaturated space between traD and traI where unidentified tra genes may be located and leaves insufficient space between traI and IS3 for coding the 94-kilodalton protein previously thought to be the product of traZ. We found that the 94-kilodalton protein arose from a translational restart and corresponds to the carboxy terminus of traI; we named it TraI*. The precise physical location of the traZ gene and the identity of its product are unknown. The oriT nicking activity known as TraZ may stem from unassigned regions between traD and traI and between traI and IS3, but a more interesting possibility is that it is actually a function of traI. On our revised map, the position of a previously detected RNA polymerase-binding site corresponds to a site at the amino terminus of traI rather than a location 1 kilobase into the coding region of the gene. Furthermore, the physical and genetic comparison of the F traD and traI genes with those of the closely related F-like conjugative plasmids R1 and R100 is greatly simplified. The translational organization we found for traI, together with its identity as the structural gene for DNA helicase I, suggests a possible functional link to several other genes from which translational restart polypeptides are expressed. These include the primases of the conjugative plasmids ColI and R16, the primase-helicase of bacteriophage T7, and the cisA product (nickase) of phage phi X174. PMID- 3036782 TI - Cloning, DNA sequence, and expression of the Rhodobacter sphaeroides light harvesting B800-850-alpha and B800-850-beta genes. AB - Two deoxyoligonucleotide probes were synthesized in accordance with the available amino acid sequence of the B800-850-beta polypeptide from Rhodobacter sphaeroides and were used to isolate a 2.6-kilobase PstI fragment from R. sphaeroides 2.4.1 chromosomal DNA. Identification of the B800-850-beta and B800-850-alpha structural genes, pucB and pucA, was confirmed by DNA sequencing. Northern (RNA) blot analysis, using restriction endonuclease fragments from the cloned genes as probes, revealed a single puc-operon-specific, highly stable transcript of approximately 640 bases present in photosynthetically grown cells. In vitro transcription-translation analysis of the puc operon revealed that the maximum synthesis of the puc operon gene products was achieved when the entire 2.6 kilobase PstI fragment was used as the template, although a 537-base-pair XmaIII fragment was sufficient to direct the synthesis of pucB and pucA fusion product. PMID- 3036781 TI - A genetic locus essential for formate-dependent growth of Bradyrhizobium japonicum. AB - A genetic locus essential for the formate-dependent growth of Bradyrhizobium japonicum was isolated by complementation of ethyl methanesulfonate-induced mutants with a cosmid gene library of B. japonicum DNA. Three related cosmids containing 18.7 kilobase pairs of B. japonicum DNA in common were identified as being able to restore formate-dependent growth capability to mutants lacking either ribulosebisphosphate carboxylase or both ribulosebisphosphate carboxylase and phosphoribulokinase activities. To further localize the complementing gene(s), a series of four deletions spanning a total of 16.1 kilobase pairs were introduced into the B. japonicum chromosome. Each resulting deletion mutant lacked formate dehydrogenase activity and lacked ribulosebisphosphate carboxylase activity and immunologically detectable protein. Three of the four also lacked phosphoribulokinase activity. Two other mutants in which the deletion-bearing recombinant plasmid had integrated into the chromosome also lacked ribulosebisphosphate carboxylase activity and protein and phosphoribulokinase activities. The genetic locus defined by these mutants could contain the structural genes for these enzymes or a regulatory gene(s) controlling their expression or both. PMID- 3036779 TI - Site-specific methylases induce the SOS DNA repair response in Escherichia coli. AB - Expression of the site-specific adenine methylase HhaII (GmeANTC, where me is methyl) or PstI (CTGCmeAG) induced the SOS DNA repair response in Escherichia coli. In contrast, expression of methylases indigenous to E. coli either did not induce SOS (EcoRI [GAmeATTC] or induced SOS to a lesser extent (dam [GmeATC]). Recognition of adenine-methylated DNA required the product of a previously undescribed gene, which we named mrr (methylated adenine recognition and restriction). We suggest that mrr encodes an endonuclease that cleaves DNA containing N6-methyladenine and that DNA double-strand breaks induce the SOS response. Cytosine methylases foreign to E. coli (MspI [meCCGG], HaeIII [GGmeCC], BamHI [GGATmeCC], HhaI [GmeCGC], BsuRI [GGmeCC], and M.Spr) also induced SOS, whereas one indigenous to E. coli (EcoRII [CmeCA/TGG]) did not. SOS induction by cytosine methylation required the rglB locus, which encodes an endonuclease that cleaves DNA containing 5-hydroxymethyl- or 5-methylcytosine (E. A. Raleigh and G. Wilson, Proc. Natl. Acad. Sci. USA 83:9070-9074, 1986). PMID- 3036783 TI - Coordinate regulation of phospholipid biosynthesis by serine in Saccharomyces cerevisiae. AB - The addition of L-serine to inositol-containing growth medium repressed membrane associated CDPdiacylglycerol synthase (CTP:phosphatidate cytidylyltransferase, EC 2.7.7.41) and phosphatidylserine synthase (CDPdiacylglycerol:L-serine O phosphatidyltransferase, EC 2.7.8.8) activities and subunit levels in wild-type Saccharomyces cerevisiae. Enzyme activities and subunit levels were not repressed when inositol was absent from the growth medium. The addition of L-serine to the growth medium did not affect the phospholipid composition of wild-type cells. CDPdiacylglycerol synthase and phosphatidylserine synthase were not regulated in the S. cerevisiae inositol biosynthesis ino2, ino4, and opi1 regulatory mutants, suggesting that regulation by inositol plus L-serine is coupled to inositol synthesis. Inositol and L-serine did not affect the activities of purified CDPdiacylglycerol synthase and phosphatidylserine synthase. The addition of compounds structurally related to L-serine to the growth medium of wild-type cells also resulted in a repression of CDPdiacylglycerol synthase and phosphatidylserine synthase but only in the presence of inositol. Phosphatidylinositol synthase (CDPdiacylglycerol:myo-inositol 3 phosphatidyltransferase, EC 2.7.8.11) was not regulated by inositol plus L serine. PMID- 3036785 TI - Tn5 insertion mutants of Pseudomonas aeruginosa deficient in surface expression of ferripyochelin-binding protein. AB - Transposon (Tn5) insertion mutants were isolated in Pseudomonas aeruginosa PAO. These mutants were screened for expression of the ferripyochelin-binding protein with monoclonal antibody in a whole-cell immunoblot assay. Fourteen mutants were identified which did not express ferripyochelin-binding protein on the cell surface. These mutants did not take up 59Fe-labeled pyochelin and grew slowly in the presence of iron chelators. PMID- 3036784 TI - Primary structure of colicin M, an inhibitor of murein biosynthesis. AB - The DNA sequence of the colicin M activity gene cma was determined. A polypeptide consisting of 271 amino acids was deduced from the nucleotide sequence. The amino acid sequence agreed with the peptide sequences determined from the isolated colicin. The molecular weight of active colicin M was 29,453. The primary translation product was not processed. In the domain required for uptake into cells, colicin M contained the pentapeptide Glu-Thr-Leu-Thr-Val. A similar sequence was found in all colicins which are taken up by a TonB-dependent mechanism and in outer membrane receptor proteins which are constituents of TonB dependent transport systems. The structure of colicin M in the carboxy-terminal activity domain had no resemblance to the pore-forming colicins or colicins with endonuclease activity. Instead, the activity domain contained a sequence which exhibited homology to the sequence around the serine residue in the active site of penicillin-binding proteins of Escherichia coli. The colicin M activity gene was regulated from an SOS box upstream of the adjacent colicin B activity gene on the natural plasmid pColBM-Cl139. PMID- 3036786 TI - Overexpression and DNA-binding properties of the mer-encoded regulatory protein from plasmid NR1 (Tn21). AB - In plasmid NR1 the expression of genes involved in mercury resistance (Tn21) is regulated by the trans-acting product of the merR gene. An in vivo T7 RNA polymerase-promoter overexpression system was used to detect a protein of approximately 16,000 daltons encoded by the merR reading frame. Overexpressed MerR constituted about 5% of labeled proteins. An in vitro MerR-mer-op (mer-op is the mer operator and promoter region) gel electrophoresis binding assay established that the binding site for MerR was located between the putative -35 and -10 sequences of the promoter for the mer structural genes. A nonsense mutation in the carboxyl half of MerR resulted in the loss of biological function and the loss of in vitro mer-op binding properties. PMID- 3036787 TI - Neutrophils become refractory to phorbol myristate acetate when treated with Ca2+ ionophore and Ca2+. AB - Human neutrophils deprived of divalent cations by treatment with ionophore A23187 in the presence of ethylene glycol bis(beta-aminoethylether)-N,N,N',N' tetraacetic acid (EGTA) showed superoxide release when they were preincubated with calcium and then treated with the ionophore. The release was not observed when the ionophore was added first and then calcium was added more than 5 min later. The absence of the release in this case can be ascribed to a refractoriness of the cells to stimuli, because the cells did not release superoxide on stimulation with phorbol myristate acetate (PMA). The cells pretreated with either calcium or the ionophore alone did release superoxide on addition of PMA. The refractoriness of the cells to PMA depended on the concentrations of calcium and the ionophore and on the time interval between the two treatments. Calcium could be replaced with Cd2+ but not with Mg2+, Ba2+, or Sr2+. The release of granular enzymes was observed when the depleted cells were pretreated with the ionophore and then with calcium. These observations indicate that calcium has dual effects on the superoxide release of neutrophils, i.e., it stimulates the cells and makes them refractory to stimuli, depending on the time interval after the addition of the ionophore, and it also regulates the enzyme release by a different mechanism. PMID- 3036788 TI - Distinction in the mode of receptor-mediated endocytosis between high density lipoprotein and acetylated high density lipoprotein: evidence for high density lipoprotein receptor-mediated cholesterol transfer. AB - The interactions of high density lipoprotein (HDL) and acetylated high density lipoprotein (acetyl-HDL) with isolated rat sinusoidal liver cells have been investigated. Cellular binding of 125I-acetyl-HDL at 0 degrees C demonstrated the presence of a specific, saturable membrane-associated receptor. This receptor was affected neither by formaldehyde-treated albumin nor by low density lipoprotein modified either by acetylation or malondialdehyde, ligands known to undergo receptor-mediated endocytosis by the cells, indicating that the receptor for acetyl-HDL constitutes a distinct class among the scavenger receptors for chemically modified proteins. Parallel binding experiments using 125I-HDL also revealed the presence on these cells of a receptor for unmodified HDL. The ligand specificities of these two receptors were similar to each other except that the acetyl-HDL receptor was sensitive to polyanions such as dextran sulfate and fucoidin. Interaction of HDL with the cells at 37 degrees C was totally different from that of acetyl-HDL. Cellular binding of HDL was not accompanied by subsequent intracellular degradation of its apoprotein moiety, whereas its cholesterol moiety was significantly transferred to the cells. In contrast, acetyl-HDL was endocytosed and underwent lysosomal degradation as a holoparticle. This shift in receptor-recognition from the HDL receptor to the acetyl-HDL receptor was accomplished by acetylation of approximately 8% of the total lysine residues of HDL apoprotein. This unique difference in endocytic behavior between HDL and acetyl-HDL suggests a potential link of the HDL receptor to HDL-mediated cholesterol transfer in sinusoidal liver cells. PMID- 3036789 TI - Detection of radical species in mixtures of some retinoids with hematin by using the ESR spin-trapping technique. AB - Radical species were detected in mixtures of some retinoids with hematin by using the ESR spin-trapping technique. The rates of radical formation were approximately proportional to the oxygen consumption during the incubation of the retinoids with hematin. HPLC analyses of the incubation mixtures of the retinoids with hematin showed that 5,6-epoxides of the retinoids were formed. The amounts of the epoxides formed were proportional to both oxygen consumption and the amounts of radicals formed. These results suggest that the 5,6-epoxidations proceed via radical intermediates. PMID- 3036790 TI - Interaction of sodium and potassium ions with Na+, K+-ATPase. III. Cooperative effect of ATP and Na+ on complete release of K+ from E2K. AB - The effects of Na+ and ATP on the K+ binding to Na+, K+-ATPase were investigated by the centrifugation method with radioactive K+ in the absence of Mg2+. In the presence of 10 microM 43KCl, 0.6 and 10 mM Na+ decreased the amount of bound K+ to one-half and zero, respectively. On the other hand, 10 microM and 10 mM ATP decreased the amount of K+ to 60 and 25-40%, respectively. When the combined effect of ATP and Na+ was tested, 10 microM ATP decreased the Na+ concentration giving half-maximal inhibition of the K+ binding to one-third, showing synergistic inhibition by both ligands, though increase in ATP concentration seemed to depress the inhibitory effect of Na+. The synergistic inhibition by ATP and Na+ suggests that the release of K+ from E2K is not completed by the binding of ATP alone but is completed by the binding of Na+ in addition to ATP during the cycle of Na+, K+-dependent ATP-hydrolysis as well as ion-transport. PMID- 3036791 TI - Determination of subunit molecular weights of canine renal (Na+,K+)-ATPase by low angle laser light scattering coupled with high performance gel chromatography in the presence of sodium dodecyl sulfate. AB - Subunit molecular weights of the (Na+,K+)-ATPase of canine renal outer medulla were estimated in the presence of sodium dodecyl sulfate (SDS) by the measuring system consisted of components connected in the following sequence: a TSK-gel G3000 SW column, a UV spectrophotometer, a low-angle laser light scattering photometer, and a differential refractometer. Polypeptide molecular weights of the alpha- and beta-subunits were determined to be 117,900 and 39,400, respectively. The measurement required the extinction coefficient at 280 nm of the sample polypeptide in addition to the outputs from the three detectors. The extinction coefficients at 280 nm of the alpha- and beta-subunits were determined to be 0.931 and 1.41 ml X mg-1 X cm-1, respectively by the quantitative amino acid analysis. The above procedure seems to be most appropriate to determine uniquely the composition of subunits molecular weights of an oligomeric membrane protein. PMID- 3036792 TI - A proton and carbon 13 nuclear magnetic resonance study of neomycin B and its interactions with phosphatidylinositol 4,5-bisphosphate. AB - The entire proton NMR spectrum of the aminoglycoside antibiotic neomycin B has been assigned at physiological pH by a combination of two-dimensional J-resolved and J-correlated and nuclear Overhauser enhancement difference spectroscopy. Unambiguous assignment of all four ring systems is possible without recourse to model or derivative compounds by observing nuclear Overhauser enhancements between as well as within rings. The subsequent assignment of the carbon 13 spectrum is simply achieved using two-dimensional heteronuclear J-correlated techniques. The proton NMR spectrum of a sonicated aqueous dispersion of the intracellular second messenger precursor phosphatidylinositol 4,5-bisphosphate is reported for the first time. The spectrum is consistent with a high degree of side chain unsaturation and a conformation for the myo-inositol head group, which appears highly mobile, in which all bulky substituents are equatorial (except the 2-hydroxyl). Addition of aliquots of phosphatidylinositol 4,5-bisphosphate to an aqueous buffered solution of neomycin B induces complex changes in the whole spectrum of the latter, including downfield shifts of differential magnitude for several well-resolved signals, viz. the anomerics, and the pair of methylene protons of the substituted cyclohexane. The complexation kinetics are fast on the NMR time scale at 25 degrees C. The binding results are discussed in terms of a tentative complexation geometry. PMID- 3036793 TI - Structure and expression of the human apolipoprotein A-IV gene. AB - We have isolated the human apolipoprotein (apo) A-IV gene from a cosmid library and determined its complete nucleotide sequence. The gene contains three exons of 162, 127, and 1180 nucleotides separated by two introns of 357 and 777 nucleotides. A sequence polymorphism has been identified in the 3' noncoding portion of the third exon. The human apoA-IV gene lacks an intron in the area encoding the 5' nontranslated region of its mRNA, which distinguishes it from all the other human apolipoprotein genes whose sequences are known. Comparison matrix analysis of the human apoA-IV gene sequence revealed evidence for an ancestral 11 nucleotide repeat unit that spans the third exon. These repeated sequences are much more highly conserved than those present in either rat apoA-IV or in any other human apolipoprotein. Optimal alignments of the 5' flanking regions of the rat and human apoA-IV genes disclosed multiple deletions in the rat sequence as well as a highly conserved region of 90 nucleotides (90% sequence identity) located within 170 nucleotides of the start site of transcription. The 5' flanking regions of the human and rat apoA-IV genes were ligated to the bacterial chloramphenicol acetyltransferase gene, then transfected into different cultured cells. The apoA-IV gene sequences elicited preferential expression of chloramphenicol acetyltransferase activity when introduced into intestinally derived Caco-2 cells and liver-derived Hep-G2 cells, consistent with the tissue specificity of the native gene. Analysis of deletion mutants of the human apoA-IV 5' flanking region indicated that regions from -293 to -233 and from -127 to -60 upstream of the transcription start site contain sequences required for maximum gene expression. These findings on the structure and expression of rat and human apoA-IV should prove useful in studying the control of the apoA-IV gene. PMID- 3036794 TI - Mouse glandular kallikrein genes. Structure and partial sequence analysis of the kallikrein gene locus. AB - Mouse glandular kallikreins are encoded by a family of closely linked genes which are located on chromosome 7 at a site corresponding to the genetically defined Tam-1, Prt-4, and Prt-5 loci. We have characterized 24 kallikrein genes by genomic cloning and restriction mapping of 310 kilobase pairs of BALB/c mouse DNA. Most of these genes are highly homologous, have the same exon/intron organization, and are linked in clusters of up to 11 genes. Partial sequence analysis of the kallikrein genes has facilitated identification of those members of the family for which protein sequence data exist and assignment of those which are pseudogenes or encode proteins of unknown function. We find that a maximum of 14 mouse kallikrein genes have the potential to encode functional proteins. PMID- 3036795 TI - Structure and organization of the murine band 3 gene. AB - The Band 3 protein mediates the reversible exchange of chloride and bicarbonate anions across the plasma membrane of erythrocytes, and probably, certain epithelial cells. It also serves to anchor the spectrin cytoskeleton to the plasma membrane via its association with ankyrin. We have isolated and largely sequenced the 17-kilobase murine Band 3 gene. We show that this gene is present in a single copy in the mouse genome and have identified and mapped the 19 intervening sequences. The locations of the intron/exon junctions in the Band 3 mRNA correlate with predicted structural features of the erythrocyte Band 3 protein structure and membrane topology. One of the introns within this gene contains a single copy of a murine Alu-type high dispersed sequence, in addition to several unusual tandemly repeated sequences. PMID- 3036796 TI - Role of the hinge protein in the electron transfer between cardiac cytochrome c1 and c. Equilibrium constants and kinetic probes. AB - A role of the hinge protein is studied in the electron transfer reaction between cytochromes c1 and c, using highly purified "one-band" cytochrome c1 and "two band" cytochrome c1. The results show that the hinge protein (Hp), which is essential for a stable ionic strength-sensitive c1-Hp-c complex, seems to play a certain role in electron transfer between cytochromes c1 and c; Keq for electron transfer reaction between cytochromes c1 and c in the presence of the hinge protein is found to be about 40% higher than that in the absence of the hinge protein at low ionic strength, but no difference exists at high ionic strength. We propose a hypothesis that the hinge protein may function as regulator for the electron transfer reaction between cytochromes c1 and c, and this may be at least one of the roles of the hinge protein in mitochondria. PMID- 3036797 TI - The chicken vimentin gene. Nucleotide sequence, regulatory elements, and comparison to the hamster gene. AB - Here we report the nucleotide sequence of the chicken vimentin gene and its deduced primary amino acid sequence. A comparison of this gene to other intermediate filament protein genes demonstrates that both exon size and position are strongly conserved features of this multigene family. In addition, the hamster and chicken vimentin genes exhibit strong identity at the level of nucleotide (74%) and amino acid (80%) sequence. Interestingly, 40% of total sequence diversity is localized to the N terminus or "head" region of these genes whereas other protein domains (rod and C terminus) are remarkably identical in both nucleotide (81%) and amino acid (89%) sequence. Even stronger amino acid identity (100%) is exhibited in certain subdomains which may define regions crucial for filament formation and function. Not surprisingly, vimentin is more homologous across animal species than it is to other intermediate filament protein members (e.g. desmin) within the same species. A comparison of 5' flanking sequences of the hamster and chicken genes as well as other characterized promoter elements (SV40, HSV-TK) reveals homologous sequence elements which may define common and/or unique sites involved in the modulation of gene expression. The implications of these sequence elements for both tissue specific and developmental expression of the vimentin gene are discussed. PMID- 3036798 TI - A new affinity matrix for mineralocorticoid receptors. AB - The behavior of mineralocorticoid and glucocorticoid receptors of rabbit kidney cytosol was investigated on two affinity gels: a new affinity matrix prepared with a 3-O-derivative of carboxymethyloxime deoxycorticosterone (deoxycorticosterone gel) and a gel linked to a 17 beta-dexamethasone derivative (dexamethasone gel). Deoxycorticosterone gel was highly specific, since it retained mineralocorticoid but not glucocorticoid receptors, and dexamethasone gel exhibited high selectivity for glucocorticoid receptors since it did not bind mineralocorticoid receptors. The use of these two matrices allowed separation of mineralocorticoid and glucocorticoid receptors and further characterization of each type of cytosolic receptors after its isolation. Cytosolic mineralocorticoid and glucocorticoid receptors stabilized by tungstate were found to have a Stokes radius of approximately 6 nm, as determined by high performance size exclusion chromatography and a sedimentation coefficient of approximately 9 S, determined on a glycerol density gradient containing tungstate, under either high or low salt conditions. The hydrodynamic parameters, binding characteristics, and specificity of mineralocorticoid receptors were the same in the untreated and dexamethasone gel-treated cytosol. Similarly glucocorticoid receptor characteristics remained unchanged after deoxycorticosterone gel treatment, indicating biochemical independence of cytosolic mineralocorticoid and glucocorticoid receptors. The [3H]aldosterone receptor complex eluted from deoxycorticosterone gel was recovered with a 30-40% yield and a purification factor of about 1000. Purified mineralocorticoid receptors had the same sedimentation coefficient as cytosolic mineralocorticoid receptors (9 S) but a different Stokes radius (4 versus 6 nm). The decrease in the Stokes radius of the purified mineralocorticoid receptors was probably due to the gel filtration method. These results indicate that the newly synthesized matrix specific for mineralocorticoid receptors constitutes a powerful tool for their extensive purification. PMID- 3036799 TI - RNA N-glycosidase activity of ricin A-chain. Mechanism of action of the toxic lectin ricin on eukaryotic ribosomes. AB - The modification reaction of 28 S rRNA in eukaryotic ribosomes by ricin A-chain was characterized. To examine whether ricin A-chain release any bases from 28 S rRNA, rat liver ribosomes were incubated with a catalytic amount of the toxin, and a fraction containing free bases and nucleosides was prepared from the postribosomal fraction of the reaction mixture by means of ion-exchange column chromatography. Thin-layer chromatographic analysis of this fraction revealed a release of 1 mol of adenine from 1 mol of ribosome. When the ribosomes or naked total RNAs were treated with ricin A-chain in the presence of [32P] phosphate, little incorporation of the radioactivity into 28 S rRNA was observed, indicating that the release is not mediated by phosphorolysis. Thus, considering together with the previous result (Endo, Y., Mitsui, K., Motizuki, M., and Tsurugi, K. (1987) J. Biol. Chem. 262, 5908-5912), the results in the present experiments demonstrated that ricin A-chain inactivates the ribosomes by cleaving the N glycosidic bond of A4324 of 28 S rRNA in a hydrolytic fashion. PMID- 3036800 TI - Complete nucleotide sequence of cDNA and predicted amino acid sequence of rat acyl-CoA oxidase. AB - cDNA clones for rat acyl-CoA oxidase were isolated. The 3.8-kilobase mRNA sequence of the enzyme was completely covered by two overlapping clones. The composite cDNA sequence consisted of 3741 bases and contained a 1983-base open reading frame which encodes a polypeptide of 661 amino acid residues. Two species of acyl-CoA oxidase cDNA were identified. They differed in their coding nucleotide sequences, only within a small region. They contained the same number of nucleotides and can be translated in a common reading frame. They are 55% and 50% homologous in the above region at the nucleotide and the amino acid levels, respectively. Both types of cDNA were isolated from a library constructed from mRNA of a single rat, thereby suggesting the occurrence of two species of acyl CoA oxidase in each rat. The amino terminus of the enzyme was determined to be N acetylmethionine, which corresponds to the initiator methionine, thus confirming the absence of a terminal presequence. We reported previously that a purified preparation of the enzyme contained three polypeptide components, A, B, and C, and suggested that components B and C are produced by a proteolytic cleavage of component A (Osumi, T., Hashimoto, T., and Ui, N. (1980) J. Biochem. (Tokyo) 87, 1735-1746). We located components B and C on the amino- and the carboxyl-terminal sides of component A. Possible functional significances of several stretches of amino acids of the enzyme are discussed, based on the sequence comparison data between rat and yeast acyl-CoA oxidases. PMID- 3036801 TI - Isolation and structural characterization of the rat acyl-CoA oxidase gene. AB - Overlapping genomic clones containing the entire sequence of the rat acyl-CoA oxidase gene were isolated and characterized. This gene spans about 25 kilobases, and there are 14 exons and 13 introns. All of the exon-intron junction sequences agree with the GT/AG rule. The results of Southern blot analyses indicated that the gene occurs once per haploid genome. In a preceding paper (Miyazawa, S., Hayashi, H., Hijikata, M., Ishii, N., Furuta, S., Kagamiyama, H., Osumi, T., and Hashimoto, T. (1987) J. Biol. Chem. 262, 8131-8137), we described the presence of two species of acyl-CoA oxidase mRNA which have different sequences, only in a small region. We identified two exons corresponding to this region. The above two mRNA species are produced by alternative splicing of these exons. Several mRNA cap sites were detected by S1 mapping and by the primer extension method. The principal one was mapped at 75 nucleotides upstream of the initiator methionine codon. The 5'-flanking region of the acyl-CoA oxidase gene was sequenced up to about 1300 nucleotides, upstream of the cap sites. There was neither a "TATA" box nor a typical "CCAAT" box sequence, at least up to nucleotide -500, rather, this region is G + C-rich (65%). "GC" box hexanucleotides (GGGCGG or CCGCCC) repeat six times in this region. Two of them, located nearest the cap sites, are in a complete 11-base pair direct repeat, 5'-GGGCGGGGCCG-3'. The features of the 5' flanking sequence of this gene resemble those of the gene for rat enoyl-CoA hydratase:3-hydroxyacyl-CoA dehydrogenase bifunctional enzyme. The possible functional significance of the characteristic 5'-flanking sequence elements in the transcriptional regulation of acyl-CoA oxidase is given attention. PMID- 3036802 TI - Structural organization of the gene for rat enoyl-CoA hydratase:3-hydroxyacyl-CoA dehydrogenase bifunctional enzyme. AB - Enoyl-CoA hydratase:3-hydroxyacyl-CoA dehydrogenase bifunctional enzyme is a monomeric protein which catalyzes the second and the third reactions of the peroxisomal beta-oxidation system. We cloned the gene for this enzyme from rat genomic libraries. The gene spans about 31 kilobases and consists of seven exons and six introns. The transcription initiation site was located 24 nucleotides upstream of the initiator methionine codon, ATG, determined by S1 nuclease mapping and primer extension analysis. The 5'-flanking region of the gene lacks typical TATA and CCAAT sequences, but contains G + C-rich sequences, including one CCGCCC ("GC" box) and two related GC hexanucleotides. Some of the structural features of the 5'-flanking region of this gene are shared by the 5'-upstream sequence of the gene for acyl-CoA oxidase, which catalyzes the first reaction of the peroxisomal beta-oxidation system and is induced coordinately with the bifunctional enzyme. Southern blot analysis of rat genomic DNA indicates that the bifunctional enzyme gene occurs once per haploid genome. The amino acid sequence of the carboxyl-terminal region of the bifunctional enzyme exhibits a significant level of homology to the sequence of pig mitochondrial 3-hydroxyacyl-CoA dehydrogenase. This region of the bifunctional enzyme is encoded mainly by the last exon (Exon VII) which codes for as much as 58% of the protein sequence. Based on the data plus our unpublished findings (N. Ishii, T. Osumi, and T. Hashimoto, unpublished data), we propose that the amino-terminal domain, which is principally encoded by Exons I-V, has enoyl-CoA hydratase activity, and the carboxyl-terminal one, which is mainly coded for by Exon VII, has a 3-hydroxyacyl CoA dehydrogenase function. PMID- 3036803 TI - Structural analysis of cDNA for rat peroxisomal 3-ketoacyl-CoA thiolase. AB - cDNA clones of rat peroxisomal 3-ketoacyl-CoA thiolase were isolated. By blotting analysis using the cDNAs as probes, the mRNA for this enzyme was estimated to be about 1.9-kilobase pairs. Elevation of mRNA levels in the liver with administration of di(2-ethylhexyl)phthalate was also evident. Sequencing analysis revealed 1,272 bases of the open reading frame which encoded 424 amino acid residues. Amino acid sequence data on six tryptic peptides and the amino terminus of the purified enzyme confirmed the cDNA sequence. The precursor of peroxisomal thiolase contains at its amino terminus a peptide extension of 26 residues. The mature enzyme is composed of 398 amino acids and the molecular weight is 41,074. The presequence has a net positive charge, lacks a long stretch of hydrophobic residues, and contains a cluster of serine residues. When the primary structure of the precursor was compared to structure of known peroxisomal proteins, there was no common homologous sequence. Peroxisomal thiolase exhibits a significant sequence homology with the mitochondrial thiolase. Possible location of the transport signal of the peroxisomal thiolase is discussed. Acyl-CoA binding sites were also located on primary structures of the two thiolases. The occurrence of interrupting sequences in several clones likely originates from intron sequences. PMID- 3036804 TI - Stimulation of insulin-like growth factor II receptor binding and insulin receptor kinase activity in rat adipocytes. Effects of vanadate and H2O2. AB - Autophosphorylation of the insulin receptor on tyrosine residues and activation of the endogenous insulin receptor kinase is postulated to be a critical step in the mechanism of action of insulin. To investigate this hypothesis, the insulin mimicking effects of vanadate (sodium orthovanadate) and H2O2 (hydrogen peroxide) alone and in combination were examined in freshly isolated rat adipocytes. Vanadate and H2O2 stimulated the translocation of insulin-like growth factor II (IGF-II) receptors to the plasma membrane of rat adipocytes in a manner analogous to insulin. IGF-II binding was increased by maximally effective doses of vanadate (1 mM), H2O2 (1 mM), and insulin (10 ng/ml) to 172 +/- 10, 138 +/- 12, and 289 +/ 16% of control, respectively. Previously (Kadota, S., Fantus, I. G., Hersh, B., and Posner, B. I. (1986) Biochem. Biophys. Res. Commun. 138, 174-178), we showed that the combination of these concentrations of vanadate plus insulin was not more potent than insulin alone. In this study, similar results were found with H2O2 plus insulin. In contrast, the combination of vanadate plus H2O2 was synergistic, effecting an increase of IGF-II binding to 488 +/- 23% of control. Amiloride inhibited the effects of vanadate, H2O2, and insulin. Adipocyte insulin receptors purified by wheat germ agglutinin chromatography were assayed for tyrosine kinase activity using the synthetic substrate poly(Glu,Tyr) (4:1). Basal activity (no in vitro insulin) was stimulated by exposure of intact cells to vanadate, H2O2, insulin, and vanadate + H2O2 to 147.7 +/- 4.3, 178.2 +/- 43.4, 495.0 +/- 67.1, and 913.2 +/- 92.0% of control, respectively. The stimulation of tyrosine kinase activity by these agents was accounted for by the insulin receptor as the augmented activity was completely immunoprecipitated with insulin receptor antibody. In these studies, the increase in IGF-II binding correlated significantly with the activation of the insulin receptor-tyrosine kinase (r = 0.927, p less than 0.001). These data support the hypothesis that activation of the insulin receptor kinase is linked to insulin action. PMID- 3036806 TI - Phosphorylation of alpha-tubulin carboxyl-terminal tyrosine prevents its incorporation into microtubules. AB - Insulin receptor kinase phosphorylated tubulin in an insulin-dependent fashion. Two different populations of phosphotubulin were found. In tubulin dimers containing tyrosine at the carboxyl-terminal of their alpha subunit, phosphate was incorporated in that residue, and the phosphorylated protein did not assemble into polymers. In tubulin dimers lacking this tyrosine residue, phosphate was incorporated into different tyrosine residues located in other parts of the molecule, and the phosphoprotein retained its capacity to polymerize. PMID- 3036805 TI - Affinity labeling of adenylate kinase with adenosine diphosphopyridoxal. Presence of Lys21 in the ATP-binding site. AB - Adenosine diphosphopyridoxal, the affinity labeling reagent specific for a lysyl residue in the nucleotide-binding site of several enzymes (Tagaya, M., and Fukui, T. (1986) Biochemistry 25, 2958-2964; Tamura, J. K., Rakov, R. D., and Cross R. L. (1986) J. Biol. Chem. 261, 4126-4133) was applied to adenylate kinase from rabbit muscle. Incubation of the enzyme with a low concentration of the reagent at 25 degrees C for 20 min followed by reduction by sodium borohydride resulted in rapid inactivation of the enzyme. Extrapolation to 100% loss of enzyme activity gave a value of 1.0 mol of the reagent per mol of enzyme. ADP, ATP, and MgATP almost completely protected the enzyme from inactivation, whereas AMP offered little retardation of the inactivation. Dilution of the inactivated enzyme which had not been treated with the reducing reagent led to restoration of enzyme activity. This reactivation was accelerated by ATP but not by AMP. Structural study of the labeled peptide showed that Lys21 is exclusively labeled by adenosine diphosphopyridoxal. These results suggest that the epsilon-amino group of Lys21 is located in the ATP-binding site of the enzyme, more specifically at or close to the subsite for the gamma-phosphate of the nucleotide. PMID- 3036807 TI - Cloning and characterization of a 12-gene cluster from Bacillus subtilis encoding nine enzymes for de novo purine nucleotide synthesis. AB - An approximately 16-kilobase pair region of the Bacillus subtilis chromosome at 55 degrees containing genes for de novo purine nucleotide synthesis (Piggot, P. J., and Hoch, J. A. (1985) Microbiol. Rev. 49, 158-179) was cloned. The nucleotide sequence of over 13 kilobase pairs indicates that this region contains a cluster of 12 genes, 11 of which encode enzymes that catalyze the 10 reactions for de novo purine nucleotide synthesis from 5-phosphoribosyl 1-pyrophosphate to IMP. The genes were identified by complementation of Escherichia coli pur mutants and by sequence comparisons with homologous enzymes. The cluster is likely an operon and is organized into three groups of overlapping genes followed by the last gene: purEKB-purC(orf)QLF-purMNH(J)-purD. Sequence comparisons provide evidence for homology of monofunctional purine nucleotide biosynthetic enzymes from B. subtilis with the corresponding multifunctional enzymes from yeast and Drosophila. Sequence alignment of the phosphoribosylaminoimidazole carboxylase heterodimer from B. subtilis with the monomeric enzyme from Methanobrevibacter smithii indicates an evolutionary relationship between these two enzymes. S1 nuclease analysis was used to map the mRNA 5' and 3' ends and to estimate levels of mRNA. These experiments indicate that synthesis of purine nucleotides is regulated independently by adenine and guanine nucleotides. Adenine nucleotides regulate transcription initiation. Guanine nucleotides regulate transcription by a termination-antitermination mechanism in a 242-nucleotide 5' untranslated mRNA leader region. Groups of overlapping genes, regulated at least in part by transcription termination-antitermination is likely to be a common theme for genetic organization and regulation of biosynthetic genes in this Gram-positive organism. PMID- 3036808 TI - Identification of a nuclear DNA binding protein associated with the interferon beta upstream regulatory region. AB - Nuclear protein extracts were prepared from uninduced myeloid leukemic KG-1 cells and analyzed for interferon-specific DNA binding by a gel electrophoresis DNA binding assay. A protein was detected that bound specifically to a 163-base pair upstream region of the IFN-beta promoter. A series of competition studies were performed to assess whether this was a general DNA binding protein; binding of this factor was competed from radiolabeled beta 163 fragment only by an excess of cold unlabeled beta 163 or a 67-base pair fragment (beta 67) derived from beta 163. Other promoter and enhancer transcriptional domains including 1) c-fos serum responsive element, 2) c-fos promoter, 3) SV40 enhancer, 4) IFN-alpha 1 promoter, and 5) plasmid pAT153 did not compete effectively for binding of the protein. These results indicate that this protein is not a general enhancer or promoter binding factor and may be unique to IFN-beta. Analysis of beta 67 DNA sequences revealed no homology to known recognition sequences for Sp1 or CTF transcription factors. PMID- 3036809 TI - Effects of prolipoprotein signal peptide mutations on secretion of hybrid prolipo beta-lactamase in Escherichia coli. AB - Hybrid proteins were constructed by coupling beta-lactamase to the signal sequence (plus nine amino acids) of selected mutant prolipoproteins of Escherichia coli. The mutant prolipoprotein signal peptides contained lesions in two structural domains of the signal peptide, the basic amino-terminal domain and the hydrophobic core domain. We then compared the processing and localization of the mutant prolipo-beta-lactamases to the processing and localization of the comparable mutant prolipoproteins. We show that a mutant signal sequence with an anionic amino terminus exhibits similar limitations in the processing of prolipo beta-lactamase as previously observed in prolipoprotein. Deletion of four hydrophobic residues from hydrophobic core results in a signal peptide which slowly translocates a fraction of the total mutant hybrid protein synthesized. This signal peptide was previously shown to translocate lipoprotein efficiently. Alteration of this hydrophobic core, which stimulated synthesis of mutant prolipoproteins, does not stimulate synthesis of prolipo-beta-lactamase. Finally mutations that slowed processing of prolipoprotein by affecting the proposed helical structure of the signal peptide had no significant effect on the processing of prolipo-beta-lactamase. These results suggest that the positively charged amino-terminal domain of the signal peptide has a common role in protein secretion regardless of the secretory protein. On the other hand, other domains of the signal peptide exhibit different phenotypes when the secretory protein is changed. PMID- 3036810 TI - Structural and functional analysis of a growth-regulated gene, the human calcyclin. AB - Calcyclin was originally defined as a cDNA clone (2A9) whose cognate RNA is growth-regulated and whose sequence shows strong similarities to the sequences of the S-100 protein, a calcium-binding protein, as well as to a subunit of the major cellular substrate for tyrosine kinase. Using the full-length cDNA, we have now isolated from a human genomic library several phages containing calcyclin sequences. One of the phages, ch. 28-10, contains the entire calcyclin gene, plus extensive flanking sequences. The calcyclin gene is a unique copy gene and has 3 exons. The 5' flanking sequence has been characterized, both structurally and functionally. Besides a TATA box, it contains, in the region proximate to the cap site, GC boxes and a sequence with a strong homology to the enhancer core of the SV40 promoter. Other enhancer-like elements are found scattered in both the 5' and 3' flanking regions. The proximate 5' flanking region is very active in driving the transient expression of linked reporters in transfection experiments. Finally, the calcyclin gene has been localized to the long arm of human chromosome 1, near the ski oncogene. PMID- 3036811 TI - Characterization and sequence analysis of a developmentally regulated putative cell wall protein gene isolated from soybean. AB - A cDNA clone, pTU04, which hybridizes to two different sizes of mRNA on Northern blots was isolated from soybean suspension culture cell poly(A) RNA. Northern analysis reveals that meristematic tissue produces a 1050-nucleotide mRNA while quiescent mature cells produce primarily a 1220-nucleotide mRNA homologous to pTU04. The cDNA and its corresponding genomic clone have been partially characterized. The nucleotide sequence of the gene predicts a proline-rich protein, designated SbPRP1, which contains a signal peptide sequence and 43 repeats of a sequence consisting primarily of Pro-Pro-Val-Tyr-Lys (CCA-CCA-GTT TAC-AAG). From nuclease S1 and hybrid-select translation analyses, the cDNA clone pTU04 appears to represent the mRNA for the mature tissue 1220-nucleotide RNA observed on Northern blots. Although there is no direct proof that the encoded protein is a cell wall protein, it has the properties similar to previously isolated cell wall proteins: 1) it is very basic with a high content of Pro, Tyr, and Lys; 2) it has similar hydropathic properties; and 3) its repeating unit shares sequence homology with that of more highly characterized cell wall proteins, generally termed extensin (Chen, J., and Varner, J. E. (1985) EMBO J. 4, 2145-2151; Smith, J. J., Muldoon, E. P., Willard, J. J., and Lamport, D. T. A. (1986) Phytochemistry 25, 1021-1030. PMID- 3036812 TI - Purification and characterization of a 2'-phosphodiesterase from bovine spleen. AB - Interferon-induced 2',5'-oligoadenylates are transiently produced during viral infection and are believed to play a role in the interferon-mediated inhibition of replication of at least some viruses. 2',5'-Oligoadenylates must be catabolized but are resistant to degradation by most known ribonucleases. A 2' phosphodiesterase that degrades 2',5'-oligoadenylates was purified 1500-fold from a low speed homogenate of bovine spleen by precipitation at pH 5.2, ammonium sulfate fractionation, differential ultrafiltration, and successive chromatography on DEAE-Sephacel, hydroxylapatite, and a fast protein liquid chromatography Mono P column. No other 2-5A-degrading activity was observed during the purification procedure. The molecular mass of the enzyme estimated by gel filtration and sodium dodecyl sulfate-polyacrylamide gel electrophoresis is 65,000. The enzyme is distinct from bovine spleen phosphodiesterase II. The 2' phosphodiesterase cleaves 2',5'- and 3',5'-linked oligonucleotides, as well as branched oligoadenylate, A(2'pA)(3'pA), but appears to be most active on 3',5' oligoribonucleotides. The enzyme cleaves 5'-AMP from the 2' terminus of 2',5' oligoadenylates and appears to require a free 2' terminus and a 3'-oxygen on the penultimate nucleotide. Substrate length, 5'-phosphorylation, and base composition do not appear to be critical factors in determining enzyme activity. The effects of pH, Mg2+, Mn2+, EDTA, phosphate, 2-mercaptoethanol, and N ethylmaleimide are also described. This enzyme may be involved in the catabolism of the interferon-induced 2',5'-oligoadenylates and other 2',5'-linked RNAs in the cell. PMID- 3036813 TI - A monoclonal antibody that inhibits cyclic AMP binding by the Escherichia coli cyclic AMP receptor protein. AB - The monoclonal antibody (mAb) 64D1 was found to inhibit cAMP binding by the cAMP receptor protein (CRP) from Escherichia coli (Li, X.-M., and Krakow, J. S. (1985) J. Biol. Chem. 260, 4378-4383). CRP is relatively resistant to attack by the Staphylococcus aureus V8 protease, chymotrypsin, trypsin, and subtilisin whereas both mAb 64D1-CRP and cAMP-CRP are attacked by these proteases yielding N terminal core fragments. The fragment patterns resulting from proteolysis of mAb 64D1-CRP and cAMP-CRP differ indicating that the CRP in each complex is in a different conformation. The data presented indicate that the preferred conformation of the antigenic site for mAb 64D1 is present in unliganded CRP. Binding of mAb 64D1 to CRP is inhibited at high cAMP concentration. Formation of a stable cAMP-CRP-lac P+-RNA polymerase open promoter complex resistant to dissociation by mAb 64D1 occurs at a much lower cAMP concentration. The observed increase in resistance to mAb 64D1 may reflect a possible conformational change in CRP effected by contact with RNA polymerase in the open promoter complex. PMID- 3036814 TI - Characterization of a photoalkylated psoralen receptor in HeLa cells. AB - Psoralens in combination with ultraviolet light are potent modulators of epidermal cell growth and differentiation. Responsive cell types contain specific, saturable, high-affinity binding sites for the psoralens. These binding sites become covalently modified by the psoralen molecule following ultraviolet light exposure. In the present studies the psoralen receptor, labeled with [3H]8 methoxypsoralen, was visualized in the cytoplasmic and plasma membrane fractions of HeLa cells following sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The receptor had an apparent molecular mass of approximately 22,000 daltons and was shown to be sensitive to protease, but not nuclease treatment. The radiolabeled receptor could not be visualized in nuclear extracts of cells. Covalent binding of the radioligand to the receptor protein was inhibited by excess unlabeled 8-methoxypsoralen, indicating that covalent psoralen-receptor binding was saturable. In addition, the covalently modified receptor was found to persist in cells for over 5 h. The presence of a cellular protein that exhibits specific affinity for the psoralens and becomes photoalkylated by these compounds, together with previous data showing that the psoralens have direct effects on the cell surface membranes, supports our model that some of the biological effects of photoactivated psoralens are receptor mediated. PMID- 3036816 TI - Development of opiate receptors and GTP-binding regulatory proteins in neonatal rat brain. AB - The mu and delta opiate receptors present in rat brain were measured independently during postnatal development. The numbers of delta receptors were almost undetectable at birth and increased substantially during the first few weeks, whereas the numbers of mu receptors remained relatively constant. Activation of either of these receptors caused inhibition of adenylate cyclase, but inhibition coupled to mu receptors was much smaller than that associated with delta receptors at all ages. Attempts to use pertussis toxin-catalyzed ADP ribosylation as an assay for the GTP-binding proteins Ni and No were hampered by the development of an NADase with age. However, specific antibodies directed against the alpha subunits of Ni or No allowed separate quantitation of these transducer proteins. Both increased with age. No is present at levels at least 5 fold higher than Ni in the adult rat brain. The N proteins are in vast excess over receptors and as such are unlikely to be limiting factors in receptor function. The data further suggest that the number of opiate receptors present throughout neonatal development is in excess over that required for optimal function. PMID- 3036815 TI - Selective thiol group modification renders fructose-1,6-bisphosphatase insensitive to fructose 2,6-bisphosphate inhibition. AB - Limited treatment of native pig kidney fructose-1,6-bisphosphatase (50 microM enzyme subunit) with [14C]N-ethylmaleimide (100 microM) at 30 degrees C, pH 7.5, in the presence of AMP (200 microM) results in the modification of 1 reactive cysteine residue/enzyme subunit. The N-ethylmaleimide-modified fructose-1,6 bisphosphatase has a functional catalytic site but is no longer inhibited by fructose 2,6-bisphosphate. The enzyme derivative also exhibits decreased affinity toward Mg2+. The presence of fructose 2,6-bisphosphate during the modification protects the enzyme against the loss of fructose 2,6-bisphosphate inhibition. Moreover, the modified enzyme is inhibited by monovalent cations, as previously reported (Reyes, A., Hubert, E., and Slebe, J.C. (1985) Biochem. Biophys. Res. Commun. 127, 373-379), and does not show inhibition by high substrate concentrations. A comparison of the kinetic properties of native and N ethylmaleimide-modified fructose-1,6-bisphosphatase reveals differences in some properties but none is so striking as the complete loss of fructose 2,6 bisphosphate sensitivity. The results demonstrate that fructose 2,6-bisphosphate interacts with a specific allosteric site on fructose-1,6-bisphosphatase, and they also indicate that high levels of fructose 1,6-bisphosphate inhibit the enzyme by binding to this fructose 2,6-bisphosphate allosteric site. PMID- 3036817 TI - Expression in Escherichia coli of BCY1, the regulatory subunit of cyclic AMP dependent protein kinase from Saccharomyces cerevisiae. Purification and characterization. AB - The regulatory (R) subunit of cAMP-dependent protein kinase from the yeast Saccharomyces cerevisiae was expressed in Escherichia coli by engineering the gene for yeast R, BCY1, into an E. coli expression vector that contained a promoter from phage T7. Oligonucleotide-directed mutagenesis was used to create an NdeI restriction site at the natural ATG of the yeast R. This facilitated construction of the T7 expression vector so that the sequence of the protein produced was identical to the natural R subunit. Yeast R was highly expressed in a soluble form. 20 mg of purified yeast R was obtained from 4 liters of E. coli. N-terminal amino acid sequencing revealed that the expressed protein began with the natural sequence. 60% of the molecules contained an N-terminal methionine, and 40% initiated with valine, the second amino acid of yeast R. The protein produced in E. coli migrated on a sodium dodecyl sulfate-polyacrylamide gel with an Mr of 52,000. The yeast R bound 2 mol of cAMP/mol of R monomer with a Kd of 76 nM. The protein was treated with urea to remove bound cAMP. Sedimentation values before and after the urea treatment were identical (s20,w = 5.1). Addition of purified R subunit to a preparation of yeast C subunit (TPK1) rendered catalytic activity cAMP-dependent with an activity ratio of 4.6. The yeast R was autophosphorylated by yeast C to a level of 0.8 mol of phosphate/mol of R monomer. By these criteria, the R subunit produced in E. coli was structurally and functionally identical to the natural yeast R subunit and similar to mammalian type II R subunits. PMID- 3036818 TI - The kinetics of reduction of yeast complex III by a substrate analog. AB - The kinetics of reduction of the cytochrome and quinone constituents of yeast complex III by the substrate homolog Q1H2 have been measured under a variety of conditions. The maximum rates of reduction of cytochromes b and c1 and of the endogenous Q6 by Q1H2 were sufficiently fast to support the Vmax for the reduction of cytochrome c by this substrate. The absorbance at 562 nm showed an initial increase which was subsequently followed by a decrease. This decrease was synchronous with the appearance of reduced cytochrome c1 and is interpreted as reflecting the absorbance contribution of c1 at 562 nm under conditions where the steady state level of the b cytochromes is constant. Prereduction of c1 and the Fe/S cluster did not affect the initial very rapid reduction of b, but the second phase was eliminated. Antimycin abolished the very rapid rate of reduction of cytochrome b in untreated complex III and completely inhibited the reduction of cytochrome b in complex III in which c1 and the Fe/S cluster had been prereduced. However, the reduction of the endogenous quinone was essentially unaffected by these treatments. Antimycin had no effect on the reduction of c1. Funiculosin also suppressed the very rapid reduction of b while both myxothiazol and 5-n undecyl-6-hydroxy-4,7-dioxobenzothiazole did not modify this phase of the reaction; no secondary decrease in absorbance was observed in the presence of any of these inhibitors. Most of the observed kinetic changes could be reproduced by simulation of the Q-cycle; simple linear and branched schemes were unable to reproduce the data. PMID- 3036819 TI - Isolation of the hemopexin receptor from human placenta. AB - A hemopexin receptor detected in detergent-solubilized placental membranes was purified from the human placenta, using hemopexin-Sepharose affinity chromatography. The solubilized membranes exhibited binding sites of 2.77 pmol of hemopexin/mg of protein with a dissociation constant (Kd) of 6.6 X 10(-8) M. The purified receptor has a molecular weight of 80,000, determined on sodium dodecyl sulfate-gel electrophoresis. Immunoinhibition experiments using the antibody against the placental receptor revealed inhibition of binding of 125I-hemopexin to human leukemia K562 and HL 60 cells, thereby strongly supporting that the polypeptide isolated from the human placenta was the hemopexin receptor. PMID- 3036820 TI - The kinetics of reoxidation of yeast complex III. An evaluation of the Q-cycle. AB - The reoxidation of reduced yeast Complex III by oxidants believed to react with cytochrome c1 exhibited multiple phases for both cytochrome c1 and the cytochromes b; the reoxidation of cytochrome b, but not cytochrome c1, was markedly slowed by the presence of antimycin. The data are consistent with the Q cycle or any other scheme which proposes a branched path for electron transport between the cytochrome b centers and the endogenous Q6, provided certain constraints are relaxed. The reoxidation of the endogenous quinone proceeded at a rate comparable to that of the rapidly reacting cytochrome b and appeared to be complete within 100 ms. Removal of the endogenous quinone did not change the rate or extent of reoxidation of any of the heme centers, demonstrating that quinone is not required for electron transport between cytochromes b and the iron-sulfur cluster. This result is inconsistent with the requirements of the Q-cycle. Funiculosin completely inhibited the reoxidation of cytochrome b whereas the reoxidation of cytochrome c1 exhibited simple first-order kinetics in the presence of this inhibitor, implying that the iron-sulfur cluster is on the direct path of electron transfer from cytochrome b to cytochrome c1. Potent inhibition of cytochrome b oxidation was also observed with myxothiazol and mucidin. The reaction of reduced Complex III with Q1 also exhibited multiple phases in the oxidation of the cytochrome b centers; these phases were unaffected by the presence of myxothiazol. Addition of antimycin, or removal of the endogenous quinone, eliminated the rapid phases; only one of the cytochrome b centers was oxidized under these conditions. Epr showed that it is the low potential cytochrome b that is the species rapidly oxidized. PMID- 3036821 TI - Structural organization and chromosomal assignment of the parvalbumin gene. AB - The structure of the rat parvalbumin gene has been elucidated from analysis of six overlapping clones isolated from a rat lambda Charon 4A genomic library. Two of the clones were mapped in detail, and all exons were localized by Southern hybridization using fragments of a full-length parvalbumin cDNA (Epstein, P., Means, A. R., and Berchtold, M. W. (1986) J. Biol. Chem. 261, 5886-5891). The rat parvalbumin transcription unit is 15.5 kilobase pairs in length and contains four introns. The first intron divides the 5'-nontranslated region, whereas the other three interrupt coding DNA. All intron/extron boundaries were sequenced as was 377 base pairs immediately 5' from the putative transcription initiation site. The promoter region contains eukaryotic regulatory homologies to the "TATA" box at -24 and "CAAT" box at -47 and -156. In addition, two doublets consisting of 11 base pair direct repeats exist in the promoter region. Parvalbumin binds two Ca2+, whereas many other members of the same superfamily bind four. Comparison of the genes that encode these proteins provides a strong confirmation of the hypothesis that parvalbumin evolved from an ancestral gene specifying a four domain Ca2+-binding protein. The rat parvalbumin gene was also utilized to assign its human counterpart to chromosome 22 from data obtained by hybridization to DNA from a somatic cell hybrid panel. It was also used to isolate a 7.5-kilobase pair EcoRI fragment from a human chromosome 22 DNA library. PMID- 3036822 TI - Identification of a stable ubisemiquinone and characterization of the effects of ubiquinone oxidation-reduction status on the Rieske iron-sulfur protein in the three-subunit ubiquinol-cytochrome c oxidoreductase complex of Paracoccus denitrificans. AB - The ubiquinol-cytochrome c oxidoreductase (cytochrome bc1) complex from Paracoccus denitrificans exhibits a thermodynamically stable ubisemiquinone radical detectable by EPR spectroscopy. The radical is centered at g = 2.004, is sensitive to antimycin, and has a midpoint potential at pH 8.5 of +42 mV. These properties are very similar to those of the stable ubisemiquinone (Qi) previously characterized in the cytochrome bc1 complexes of mitochondria. The micro environment of the Rieske iron-sulfur cluster in the Paracoccus cytochrome bc1 complex changes in parallel with the redox state of the ubiquinone pool. This change is manifested as shifts in the gx, gy, and gz values of the iron-sulfur cluster EPR signal from 1.80, 1.89, and 2.02 to 1.76, 1.90, and 2.03, respectively, as ubiquinone is reduced to ubiquinol. The spectral shift is accompanied by a broadening of the signal and follows a two electron reduction curve, with a midpoint potential at pH 8.5 of +30 mV. A hydroxy analogue of ubiquinone, UHDBT, which inhibits respiration in the cytochrome bc1 complex, shifts the gx, gy, and gz values of the iron-sulfur cluster EPR signal to 1.78, 1.89, and 2.03, respectively, and raises the midpoint potential of the iron sulfur cluster at pH 7.5 from +265 to +320 mV. These changes in the micro environment of the Paracoccus Rieske iron-sulfur cluster are like those elicited in mitochondria. These results indicate that the cytochrome bc1 complex of P. denitrificans has a binding site for ubisemiquinone and that this site confers properties on the bound ubisemiquinone similar to those in mitochondria. In addition, the line shape of the Rieske iron-sulfur cluster changes in response to the oxidation-reduction status of ubiquinone, and the midpoint of the iron-sulfur cluster increases in the presence of a hydroxyquinone analogue of ubiquinone. The latter results are also similar to those observed in the mitochondrial cytochrome bc1 complex. However, unlike the mitochondrial complexes, which contain eight to 11 polypeptides and are thought to contain distinct quinone binding proteins, the Paracoccus cytochrome bc1 complex contains only three polypeptide subunits, cytochromes b, c1, and iron-sulfur protein. The ubisemiquinone binding site and the site at which ubiquinone and/or ubiquinol bind to affect the Rieske iron sulfur cluster in Paracoccus thus exist in the absence of any distinct quinone binding proteins and must be composed of domains contributed by the cytochromes and/or iron-sulfur protein. PMID- 3036824 TI - Inactivation of the RNA polymerase of vesicular stomatitis virus by N ethylmaleimide and protection by nucleoside triphosphates. Evidence for a second ATP binding site on L protein. AB - The purified RNA polymerase complex of vesicular stomatitis virus required added thiols for maximal activity, whereas polymerase activity from whole disrupted virions did not. Maximal activity of the purified polymerase complex required greater than or equal to 1 mM added dithiothreitol. The polymerase was inactivated by N-ethylmaleimide (NEM) at 0 degree C, with k2 = 528 +/- 26 M-1 min 1. Activity was recovered by addition of L protein, but not N or NS, to the NEM inactivated complex, indicating that the NEM-sensitive group was present on the L protein. Nucleoside triphosphates protected the enzyme against inactivation by N ethylmaleimide. ATP was most effective, with KD = 0.58 +/- 0.07 mM, a value close to the Km of ATP reported previously for initiation of RNA synthesis. dATP was nearly as effective, and GTP was slightly less effective than ATP. Non hydrolyzable analogs of ATP protected weakly, whereas ADP and pyrimidine triphosphates gave very poor, but still measurable, protection. The ATP binding site thus identified differs from the protein kinase-associated ATP binding site identified on L protein by Sanchez et al. (Sanchez, A., De, B.P., and Banerjee, A. K. (1985) J. Gen. Virol. 66, 1025-1036) in having a substantially lower affinity for ATP. Two putative ATP binding sites were identified in the L protein amino acid sequence, but none were found in the N or NS sequences. PMID- 3036823 TI - Cloning and sequencing of the ornithine decarboxylase gene from Trypanosoma brucei. Implications for enzyme turnover and selective difluoromethylornithine inhibition. AB - Ornithine decarboxylase of the African trypanosome Trypanosoma brucei brucei had an estimated native molecular weight of 100,000 by gel filtration and a subunit molecular weight of 45,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The gene encoding this enzyme, present in a single copy in T. brucei, was identified by mouse ornithine decarboxylase cDNA under relatively stringent conditions of hybridization and subcloned in a 5.9-kilobase (kb) SstI fragment from a cosmid clone into the plasmid pUC 19. This clone encompassed a 2.8-kb SstII fragment that contained the entire T. brucei ornithine decarboxylase gene. The 2.8-kb SstII fragment hybridized to a 2.4-kb mRNA that presumably encodes the parasite enzyme. The 2.8-kb SstII fragment was partially sequenced and found to contain an open reading frame of 445 amino acids that has 61.5% homology with the corresponding sequence of the mouse enzyme. The only major discrepancies between the two enzymes are the addition of a 20-amino acid N terminal peptide and the deletion of a 36-amino acid C-terminal peptide and the T. brucei ornithine decarboxylase. The C terminus has been postulated to be one of the structural factors associated with rapid in vivo turnover of mammalian ornithine decarboxylase. The absence of this C-terminal peptide in T. brucei ornithine decarboxylase predicts a slow turnover for the parasite enzyme in vivo, and this is supported by our experimental data. The lack of turnover of ornithine decarboxylase in trypanosomes may constitute the basis of selective antitrypanosomal action of the irreversible enzyme inhibitor DL-alpha difluoromethylornithine. PMID- 3036825 TI - Identification of a region in the human vasoactive intestinal polypeptide gene responsible for regulation by cyclic AMP. AB - Transcription of the vasoactive intestinal polypeptide (VIP) gene is regulated by cAMP. To identify the nucleotide sequences in the human VIP gene responsible for this regulation, we constructed chimeric genes containing different portions of the 5'-flanking region of the human VIP gene fused to the structural sequence encoding the bacterial reporter enzyme chloramphenicol acetyltransferase (CAT). The transcriptional activities of the fusion genes introduced into the rat pheochromocytoma cell line PC12 were assayed by measuring CAT activity in the cell lysates. Forskolin, an adenylate cyclase-activating agent, stimulated the expression of VIP-CAT fusion genes. Deletional analysis demonstrated that a region between -86 and -70 nucleotides upstream from the transcriptional origin of the human VIP gene was responsible for stimulation by forskolin. This region was able to confer cAMP-responsiveness to a gene that is not normally regulated by cAMP. Two copies of a 5 base pair motif, 5'-CGTCA-3', are required for activity of the VIP cAMP regulatory region. This motif is also present in the cAMP regulatory region of several other eukaryotic genes. PMID- 3036826 TI - Class II genes of the human major histocompatibility complex. Organization and evolutionary relationship of the DR beta genes. AB - The genes of the polymorphic HLA-DR molecules are located within the human major histocompatibility complex. We have studied the HLA-DR genes of an HLA homozygous individual typed to be DR4, Dw4, and DRw53. Fourteen cosmid and phage clones from genomic libraries were isolated and grouped into three clusters comprising a total of 165 kilobases. These clusters contain four DR beta genes. Nucleotide sequence determination showed that two of the genes encode beta chains that carry the DR4 and DRw53 specificities, respectively, while the other two genes are presumably pseudogenes. Comparisons of the nucleotide sequences of all four DR beta genes of the DR4 haplotype show that the genes are extensively similar, approximately 90% in both exons and introns. All four genes are equally similar to each other. These observations are consistent with the notion that the genes arose by duplications that were followed by homogenization through gene conversion. The existence of more than one DR beta gene homologue but only a single DR alpha gene homologue in mouse, rabbit, and cattle suggests that the DR beta gene duplications occurred at or early during mammalian speciation. PMID- 3036827 TI - Class II genes of the human major histocompatibility complex. The DO beta gene is a divergent member of the class II beta gene family. AB - A novel class II beta chain gene is described. This gene, tentatively called DO beta, displays considerably less polymorphism than beta genes of the DP, DQ, and DR loci. The nucleotide sequence of the DO beta gene is strikingly similar to that of the previously identified murine A beta 2 gene. The DO beta gene displays the same exon/intron organization as other beta genes although the fifth exon and the translated portion of the sixth exon are longer than in other genes. A striking feature of the amino acid sequence deduced from the DO beta gene sequence is the pronounced hydrophobicity of the NH2-terminal region. This feature distinguishes the putative DO beta chain from other class II beta chains and raises the possibility that DO beta chains may interact with an alpha chain that is structurally different from those of the DP, DQ, and DR loci. It further suggests that the putative DO molecule may have a function different from those of other class II antigens. PMID- 3036828 TI - Class II genes of the human major histocompatibility complex. Comparisons of the DQ and DX alpha and beta genes. AB - The human major histocompatibility complex, HLA, contains the genes of several class II molecules. We present here the molecular maps of the DQ and DX subregions and analyze the sequences of the polymorphic DQ alpha and DQ beta genes as well as the DX alpha and DX beta genes. The DQ alpha and DQ beta genes are oriented in opposite directions, approximately 12 kilobases apart. The DX alpha and DX beta genes are similarly oriented about 8 kilobases. The exon-intron organizations of the DQ alpha and DX alpha genes are analogous to those of other class II alpha genes. Comparison of the DQ alpha gene sequence to three DQ alpha cDNA clones shows that amino acid replacements are predominantly located between residues 45 and 80 in the amino-terminal domain. Analysis of the frequency of silent and replacement substitutions indicates that there is little selection against replacements in DQ alpha first domains. The exons encoding the second domains of DQ alpha and DX alpha are virtually identical, suggesting that a gene conversion event has occurred between these genes. The DX beta gene is very similar to the DQ beta gene but differs in the cytoplasmic portion. The DX beta gene contains a separate exon of 24 nucleotides encoding the core of the cytoplasmic tail. This exon is not expressed in the DQ beta genes due to a nonfunctional splice junction. Comparison of the number of nucleotide substitutions in the DQ beta first and second domain exons suggests that little or no phenotypic selection acts on the first domain whereas the second domain is under strong selection. PMID- 3036829 TI - Class II genes of the human major histocompatibility complex. Evolution of the DP region as deduced from nucleotide sequences of the four genes. AB - The DP region of the human major histocompatibility complex contains two alpha genes and two beta genes. The DP alpha 1 and beta 1 genes encode the expressed DP histocompatibility antigen molecule, while the DP alpha 2 and beta 2 genes are inactive in the haplotypes examined. Here we present the sequence of the two DP beta genes and of the expressed DP alpha 1 gene. Nucleotide sequence comparisons reveal a considerably greater degree of similarity between the two beta genes than between the two alpha genes. We propose that a duplication giving rise to the DP alpha gene pair evolutionarily preceded the corresponding DP beta gene duplication. We also propose, based on the orientation of other class II gene pairs, that the original DP molecule was encoded by the DP beta 1 and DP alpha 2 genes. At some stage during the evolution of the DP region both of the two pseudogenes appear to have been expressed. PMID- 3036830 TI - Nucleotide sequence of the overlapping genes for the subunits of Bacillus subtilis aspartokinase II and their control regions. AB - The nucleotide sequence of a 2.9-kilobase Bacillus subtilis DNA fragment containing the entire coding region of aspartokinase II and adjacent chromosomal regions (Bondaryk, R. P., and Paulus, H. (1985a) J. Biol. Chem. 260, 585-591) has been determined. The results confirmed the earlier prediction that the two subunits of aspartokinase II, alpha and beta, are encoded by in-phase overlapping genes. The nucleotide sequence showed strong ribosome binding sites before the translation initiation codons of the alpha and beta subunits. Deletion of most of the coding region unique to the alpha subunit had no effect on the synthesis of the smaller beta subunit, demonstrating that the beta subunit is indeed the product of independent translation. The site of transcription initiation of the aspartokinase gene was found to be more than 300 nucleotides upstream from the translation start of the alpha subunit. The intervening region contained a short reading frame capable of encoding a 24-residue lysine-rich polypeptide, which overlaps a region of extensive dyad symmetry culminating in a rho-independent transcription terminator. This region may be an attenuator control element that regulates the expression of the aspartokinase gene in response to the availability of lysine, the end product of the pathway. The coding sequence of the aspartokinase II subunits was immediately followed by a rho-independent transcription terminator. This termination site has an unusual symmetry, which allows it also to serve as transcription terminator for a gene that converges on the aspartokinase II gene from the opposite direction, an interesting example of genetic economy. The deduced amino acid sequence of B. subtilis aspartokinase II was compared with the sequences of the three aspartokinases from Escherichia coli (Cassan, M., Parsot, C., Cohen, G. N., and Patte, J. C. (1986) J. Biol. Chem. 261, 1052-1057). Significant sequence similarities suggest a close evolutionary relationship between the four enzymes. PMID- 3036832 TI - Gene 18 protein of bacteriophage T7. Overproduction, purification, and characterization. AB - Genetic and physical analyses indicate that gene 18 protein of bacteriophage T7 is essential for packaging of T7 DNA. T7 DNA is replicated via linear intermediates, culminating in the formation of concatemers many genomes in length which are then packaged into capsids. In infections with phage carrying amber mutations in gene 18, development is blocked at the concatemer stage. Biochemical studies on the role of gene 18 protein in concatemer processing and DNA packaging have been hampered by its low level of expression of gene 18 during T7 infections. We have cloned gene 18 on a plasmid downstream from the bacteriophage lambda PL promoter controlled by the temperature-sensitive lambda repressor encoded by c 1857. Thermal induction leads to the expression of the 10,000-Da gene 18 protein to the extent of approximately 10% of the total protein after 2 h. The overexpressed gene 18 protein is susceptible to proteolytic degradation, a condition that can be alleviated by expression in an Escherichia coli strain carrying the lon100 deletion which reduces production of protease La. Extracts of induced cells will complement an extract of T7-infected cells lacking gene 18 protein for packaging of exogenous T7 DNA. The assay has been used to monitor the purification of gene 18 protein to essential homogeneity. The identity of the purified protein has been confirmed by sequencing of the N terminus. Gel filtration analysis suggests that the native protein is an octomer. Treatment of gene 18 protein with 3 M guanidine hydrochloride denatures it to a monomer. Removal of the denaturing agent by dialysis regenerates the octomeric structure and the ability to complement packaging extracts. PMID- 3036831 TI - A small hydrophobic domain anchors leader peptidase to the cytoplasmic membrane of Escherichia coli. AB - Leader peptidase is an enzyme of the Escherichia coli cytoplasmic membrane which removes amino-terminal leader sequences from many secreted and membrane proteins. Three potential membrane-spanning segments exist in the first 98 amino acids of leader peptidase. We have characterized the topology of leader peptidase based on its sensitivity to protease digestion. Proteinase K and trypsin treatment of right-side-out inner membrane vesicles and spheroplasts yields protected fragments of approximately 80 and 105 amino acid residues, respectively. We have shown that both fragments are derived from the amino terminus of the protein and that the smaller protected peptide can be derived from the larger. Removal of the third potential membrane-spanning segment (residues 82-98) does not affect the size of the proteinase K-protected fragment but does reduce the size of the trypsin-protected peptide. Because the proteinase K-protected fragment is about 9000 daltons, is derived from the amino terminus of leader peptidase, and its size is not affected when amino acids 82-98 are removed from the protein, it must extend from the amino terminus to approximately residue 80. Likewise, the trypsin protected fragment must extend from the amino terminus to about residue 105. These data suggest a model for the orientation of leader peptidase in which the second hydrophobic stretch (residues 62-76) spans the cytoplasmic membrane and the third hydrophobic stretch resides in the periplasmic space. PMID- 3036833 TI - ERp99, an abundant, conserved glycoprotein of the endoplasmic reticulum, is homologous to the 90-kDa heat shock protein (hsp90) and the 94-kDa glucose regulated protein (GRP94). AB - We have isolated an expressible full-length cDNA clone encoding murine ERp99, an abundant, conserved transmembrane glycoprotein of the endoplasmic reticulum membrane. ERp99 is synthesized as a 92,475-kDa precursor containing 802 amino acids. It possesses a signal peptide of 21 amino acids which is cleaved cotranslationally. Analysis of the amino acid sequence deduced from the nucleotide sequence of the cDNA clone led us to propose a model for the orientation of ERp99 in the endoplasmic reticulum membrane. In this model, ERp99 possesses one membrane-spanning, stop transfer segment in the N-terminal region. The protein chain passes through the membrane only once, and approximately 75% of the protein remains on the cytoplasmic side of the ER membrane. Comparison of the ERp99 sequence to the sequence of other proteins revealed that ERp99 has extensive homology with the 90-kDa heat shock protein of Saccharomyces cerevisiae (hsp90) and the 83-kDa heat shock protein of Drosophila melanogaster. In addition, the N terminus of mature ERp99 is identical to that of the 94-kDa glucose regulated protein (GRP94) of mammalian cells. PMID- 3036834 TI - An internalized amino-terminal signal sequence retains full activity in vivo but not in vitro. AB - Internalization of the signal sequence of the vesicular stomatitis virus glycoprotein was accomplished by extending the amino-terminal coding sequence with sequences derived from pBR322. Such constructs were then expressed in eukaryotic cells. It was found that amino-terminal extensions consisting of 20, 61, or 102 amino acids totally unrelated to any signal peptide affected neither the function nor cleavage of the signal sequence in vivo. Subsequent transport of the glycoprotein was also not affected. Although the internalized signals functioned with wild-type efficiency in vivo, membrane insertion in vitro (as determined by proteolysis protection assays), signal cleavage, and glycosylation were only achieved when the amino-terminal presequences were short. PMID- 3036835 TI - Mutations in the NH2-terminal domain of the signal peptide of preproparathyroid hormone inhibit translocation without affecting interaction with signal recognition particle. AB - The amino-terminal domain of a eukaryotic signal peptide, from bovine parathyroid hormone, was altered by in vitro mutagenesis of the cDNA. The function of "internalized" signal sequence mutants and of deletion mutants was assayed using an in vitro translation-translocation system. The addition of 11 amino acids to the NH2 terminus of the signal peptide did not prevent normal processing of the precursor protein, whereas a 23-amino acid extension blocked processing. These data suggest that the NH2-terminal sequences of internal signal peptides must be permissive of the signal function. Deletion of 6 NH2-terminal amino acids from the signal peptide had no effect on its cleavage by microsomal membranes, but removal of 10 or 13 amino acids, including all charged residues prior to the hydrophobic core, prevented processing. For both the extension and deletion mutations, processed proteins were protected from proteolytic digestion, whereas unprocessed forms were not, which indicated that the unprocessed mutant proteins were not translocated across the microsomal membrane. Translation of both the extension and deletion translocation-deficient mutants was arrested by signal recognition particle, and salt-washed microsomal membranes reversed the translational arrest. These data demonstrate that the NH2-terminal domain is not required for the interaction of signal recognition particle with the signal peptide or with signal recognition particle receptor, but is required for formation of a maximally translocation-competent complex with the microsomal membrane. PMID- 3036836 TI - Isolation and characterization of the nuclear gene encoding the Rieske iron sulfur protein (RIP1) from Saccharomyces cerevisiae. AB - The nuclear gene encoding the Rieske iron-sulfur protein of the cytochrome bc1 complex of the mitochondrial respiratory chain has been isolated and characterized from Saccharomyces cerevisiae. We used a segment of the iron-sulfur protein gene from Neurospora crassa (Harnisch, U., Weiss, H., and Sebald, W. (1985) Eur. J. Biochem. 149, 95-99) to detect the yeast gene by Southern analysis. Five different but overlapping clones were then isolated by probing a yeast genomic library carried on YEp 13 by colony lift hybridization. Several approaches confirmed that the isolated DNA contained the gene for the Rieske iron sulfur protein. The yeast gene, which contains no introns, can be expressed in Escherichia coli. A 900-base pair HindIII-EcoRI fragment was subcloned into pUC19 and directed the synthesis of immunodetectable protein. The gene was also identified by disruption of its chromosomal copy by homologous integration. A 400 base pair PstI-EcoRI fragment cloned adjacent to a HIS3 marker in pUC18 was used as an integrating vector. HIS+ transformants were obtained which were unable to grow on the nonfermentable carbon source glycerol. Southern analysis of the respiration deficient (gly-) strains confirmed that the chromosomal copy of the gene was disrupted, and immunoblots of extracts of the transformants indicated a lack of iron-sulfur protein. A respiration-deficient integrant was transformed to GLY+ by a 2-kilobase pair HindIII-BglII fragment, including a complete copy of the gene, carried on a multicopy episomal vector. Immunoblots with monoclonal antibodies to the iron-sulfur protein indicated overproduction of the protein in the complemented strain and revealed expression of approximately equal amounts of mature iron-sulfur protein and of a protein approximately 3 kDa larger than the mature protein in the complemented strain. A 1.2-kilobase pair segment of DNA from the clone which complemented the disrupted strains was sequenced and found to contain an open reading frame of 645 nucleotides, capable of encoding a 21,946 dalton protein. The gene is flanked by consensus signal sequences for initiation and termination which are common in yeast and is preceded by a possible upstream activating sequence. Amino acid sequence analysis of the amino-terminal end of the mature iron-sulfur protein agreed exactly with that predicted by the nucleotide sequence starting at Lys-31.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3036837 TI - Identification of two isoforms of the catalytic subunit of Na,K-ATPase in myocytes from adult rat heart. AB - The present study demonstrates that two forms of the alpha catalytic subunit of the Na,K-ATPase are present in rat heart and originate from cardiomyocytes. They were resolved on sodium dodecyl sulfate-polyacrylamide gel electrophoresis after reduction and alkylation of the sulfhydryl groups. The two forms were identified on immunoblots using two specific antisera against either the alpha subunit from Bufo marinus kidney and the alpha and beta subunits from lamb kidney. Comparison of the two forms to the alkylated Na,K-ATPase from rat kidney (containing one catalytic subunit) and from rat brain (containing alpha and alpha + subunits) suggested that, in rat cardiac myocytes, the form with a fast migration rate (alpha F) corresponds to the alpha subunit of low ouabain affinity and the one with a slow migration rate (alpha S), to a subunit of high ouabain affinity. Thus, the existence of two isoforms of the catalytic subunit in cardiac myocytes accounts well for the biphasic ouabain inhibition of the Na,K-ATPase activity and for the biphasic inotropic responsiveness to cardiac glycosides of the rat heart. PMID- 3036838 TI - Multiple interconvertible affinity states for the delta opioid agonist-receptor complex. AB - Previously we demonstrated that rates of dissociation of [3H-D-Ala2-D Leu5]enkephalin [( 3H]DADL) from bovine hippocampal synaptic plasma membranes (SPMs) varied depending upon association time, suggesting a multistep binding process. To characterize different kinetic intermediates, we examined the effects of guanine nucleotide on dissociation rate. Control off-rates were compared to those obtained when guanyl-5'-yl-imidodiphosphate (Gpp(NH)p) (50 microM) was added either coincident with the radioligand at association or with 1 microM unlabeled DADL which initiated dissociation. delta site selectivity of [3H]DADL was ensured by addition of 20 nM unlabeled [D-Ala2-Me-Phe4-Gly-Ol5]enkephalin which suppressed mu site cross-reactivity in this preparation. Addition of Gpp(NH)p at the onset of dissociation increased the off-rate to a much greater extent after steady state binding was reached (60 min) compared to that following an association time of only 7 or 20 min. A slowly formed high affinity state appeared to be rapidly converted to a lower affinity state under these conditions. When Gpp(NH)p was present throughout the association period, the slowly dissociating state was no longer observed. Also, the off-rate following a 7-min association is linear and much faster than control, suggesting that Gpp(NH)p may affect an initial intermediate state as well as the high affinity complex. Pretreatment of the membranes with N-ethylmaleimide (NEM) eliminated the association time-dependent dissociation rates, apparently preventing time dependent formation of a high affinity state. This state is thought to be possibly a ligand-receptor complex interacting with a GTP binding protein. However, the rate of dissociation from NEM-treated membranes was accelerated by addition of Gpp(NH)p and the effect was not association time-dependent. NEM treatment resulted in an increased potency for Gpp(NH)p inhibition of [3H]DADL steady state binding. These results suggest the occurrence of at least three steps in the association of DADL to bovine hippocampal synaptic membranes. PMID- 3036839 TI - The interaction between the presynaptic phospholipase neurotoxins beta bungarotoxin and crotoxin and mixed detergent-phosphatidylcholine micelles. A comparison with non-neurotoxic snake venom phospholipases A2. AB - Certain phospholipase A2 enzymes (E.C.3.1.1.4) selectively inhibit neurotransmitter release from cholinergic nerve terminals. Both specific acceptor proteins and the physical state of nerve terminal phospholipids have been implicated in studies of the mechanism of phospholipase neurotoxin action. Here we have examined the effects of charge on a micellar phospholipid substrate by comparing the enzyme activity and binding of two neurotoxic phospholipases (beta bungarotoxin and crotoxin) with other non-neurotoxic phospholipases. This has been achieved by altering either the phospholipid or the ionic charge of the detergent in the mixed phospholipid micelle. The neurotoxic phospholipases were only active on negatively charged micelles, whereas the non-neurotoxic enzymes were equally active in hydrolyzing neutral micelles. This distinction was also reflected in binding studies; the non-neurotoxic phospholipases bound to both types of substrate, whereas beta-bungarotoxin and crotoxin selectively bound to negatively charged micellar structures. These experiments suggest that, in addition to the existence of any specific acceptor proteins, neurotoxin binding is also governed by the charge on the lipid phase of the nerve terminal membrane. PMID- 3036840 TI - Purification and characterization of the beta-adrenergic receptor kinase. AB - The beta-adrenergic receptor kinase (beta-ARK) is a recently discovered enzyme which specifically phosphorylates the agonist-occupied form of the beta adrenergic receptor (beta-AR) as well as the light-bleached form of rhodopsin. beta-ARK is present in a wide variety of mammalian tissues. The kinase can be purified from bovine cerebral cortex to greater than 90% homogeneity by sequential chromatography on Ultrogel AcA34, DEAE-Sephacel, CM-Fractogel, and hydroxylapatite. This results in an approximately 20,000-fold purification with an overall recovery of 12%. The purified kinase has an Mr approximately 80,000 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Several findings indicate that this peptide contains the beta-ARK activity. First, on hydroxylapatite chromatography the enzyme activity coelutes with the Mr approximately 80,000 protein as revealed by Coomassie-Blue staining. Second, under phosphorylating conditions the Mr approximately 80,000 protein is phosphorylated. Finally, the Mr approximately 80,000 protein specifically interacts with reconstituted agonist-occupied beta-AR. Kinetic parameters of the enzyme for beta-AR are Km = 0.25 microM and Vmax = 78 nmol/min/mg whereas for rhodopsin the values are Km = 6 microM and Vmax = 72 nmol/min/mg. The Km value of the enzyme for ATP is approximately 35 microM using either beta-AR or rhodopsin as substrate. Receptor phosphorylation by beta-ARK is effectively inhibited by Zn2+, digitonin and a variety of salts. The availability of purified beta-ARK should greatly facilitate studies of its role in receptor desensitization. PMID- 3036842 TI - The Cu(II)-repressible plastidic cytochrome c. Cloning and sequence of a complementary DNA for the pre-apoprotein. AB - We have cloned a complementary DNA for pre-apocytochrome c-552 from Chlamydomonas reinhardtii. The deduced sequence of the mature protein shows high homology to those of cytochromes c-553 from cyanobacteria. Its homology to mitochondrial cytochrome c or bacterial photosynthetic cytochrome c2 is lower and appears to be concentrated in sequences around amino acids involved in the interaction with heme. With respect to primary sequence, the "transit sequence" for cytochrome c 552 appears to show no homology to other transit sequences for nuclear encoded chloroplast proteins. However, based on analogy to transit sequences for other proteins (Daldal, F., Cheng, S., Applebaum, J., Davidson, E., and Prince, R. C. (1986) Proc. Natl. Acad. Sci. U.S.A. 83, 2012-2016; Goldschmidt-Clermont, M., and Rahire, M. (1986) J. Mol. Biol. 191, 421-432; Smeekens, S., de Groot, M., van Binsbergen, J., and Weisbeek, P. (1986) Cell 46, 365-375) the transit sequence of cytochrome c-552 can be divided into envelope-traversing and thylakoid-traversing domains. Cytochrome c-552 appears to encoded by a single nuclear gene in C. reinhardtii. The gene is expressed exclusively in Cu(II)-deficient cells. PMID- 3036841 TI - Turkey erythrocyte membranes as a model for regulation of phospholipase C by guanine nucleotides. AB - Phosphoinositides of human, rabbit, rat, and turkey erythrocytes were radiolabeled by incubation of intact cells with [32P]Pi. Guanosine 5'-O (thiotriphosphate) (GTP gamma S) and NaF, which are known activators of guanine nucleotide regulatory proteins, caused a large increase in [32P]inositol phosphate release from plasma membranes derived from turkey erythrocytes, but had no effect on inositol phosphate formation by plasma membranes prepared from the mammalian erythrocytes. High performance liquid chromatography analysis indicated that inositol bisphosphate, inositol 1,3,4-trisphosphate, inositol 1,4,5 trisphosphate, and inositol 1,3,4,5-tetrakisphosphate all increased by 20-30-fold during a 10-min incubation of turkey erythrocyte membranes with GTP gamma S. The increase in inositol phosphate formation was accompanied by a similar decrease in radioactivity in phosphatidylinositol 4-phosphate (PIP) and phosphatidylinositol 4,5-bisphosphate (PIP2). GTP gamma S increased inositol phosphate formation with a K0.5 of 600 nM; guanosine 5'-(beta, gamma-imido)trisphosphate was 50-75% as efficacious as GTP gamma S and expressed a K0.5 of 36 microM. Although GTP alone had little effect on inositol phosphate formation, it blocked GTP gamma S stimulated inositol phosphate formation, as did guanosine 5'-O-(2 thiodiphosphate). Turkey erythrocytes were also shown to express phosphatidylinositol synthetase activity in that incubation of cells with [3H] inositol resulted in incorporation of radiolabel into phosphatidylinositol, PIP, and PIP2. Incubation of membranes derived from [3H]inositol-labeled erythrocytes with GTP gamma S resulted in large increases in [3H] inositol phosphate formation and corresponding decreases in radiolabel in PIP and PIP2. The data suggest that, in contrast to mammalian erythrocytes, the turkey erythrocyte expresses a guanine nucleotide-binding protein that regulates phospholipase C, and as such, should provide a useful model system for furthering our understanding of hormonal regulation of this enzyme. PMID- 3036843 TI - CTP:phosphorylcholine cytidylyltransferase from rat liver. Isolation and characterization of the catalytic subunit. AB - We reported previously the purification of CTP:phosphorylcholine cytidylyltransferase from rat liver (Weinhold, P. A., Rounsifer, M. E., and Feldman, D. A. (1986) J. Biol. Chem. 261, 5104-5110). The purified enzyme appeared to contain equal amounts of two nonidentical proteins, with Mr of about 38,000 and 45,000. We have now separated and purified these proteins. Polyacrylamide electrophoresis in the presence of sodium dodecyl sulfate indicated that each protein was homogeneous. The 45,000 protein contained the catalytic activity. Analysis by gel filtration chromatography and glycerol gradient centrifugation indicated that the 38,000 and 45,000 proteins in the purified cytidylyltransferase were independently associated with Triton X-100 micelles. The apparent Mr of the complexes suggested that a tetramer of each protein was bound to one Triton X-100 micelle. The isolated 45,000 catalytic protein had the same lipid requirement and kinetic properties as the purified cytidylyltransferase containing both proteins. Enzyme activity was stimulated to maximal values by phosphatidylcholine vesicles containing 9 mol % of either oleic acid, phosphatidylinositol, or phosphatidylglycerol. The amino acid compositions of the isolated 38,000 and 45,000 proteins were distinctly different. Overall, the results suggested that a tetramer of the 45,000 protein possessed nearly optimal catalytic activity. A functional role of the 38,000 protein as part of a cytidylyltransferase enzyme complex could not be documented. However, the need for stabilizing concentrations of Triton X-100 in the purified enzyme preparation may have prevented the association of the two proteins. PMID- 3036844 TI - Purification and characterization of nicotinamide deamidase from yeast. AB - Nicotinamide deamidase (YNDase) has been purified from yeast through the use of a six-step procedure that includes molecular-sieve high performance liquid chromatography. The final preparation was homogeneous by the criteria of sodium dodecyl sulfate-gel electrophoresis, and the enzyme specific activity was determined to be 175 mumol of nicotinate formed per min/mg enzyme. Gel electrophoresis and molecular-sieve high performance liquid chromatography were employed also to characterize YNDase as a monomeric protein with a molecular weight of 34,000. A Km value for nicotinamide of 33 microM was determined for the deamidase activity at pH 6, and a pH range for optimal stability of 6-8.5 was established for this enzyme. The YNDase activity was also examined over a pH range at several substrate concentrations and both the log Vmax and log Vmax/Km plots versus pH suggested that a protonated amino acid residue with an apparent pKb value of 7.8 was essential to this activity. During an in vitro assay of the YNDase-catalyzed formation of nicotinate, ammonia was generated and detected chemically. Inhibition of the YNDase activity by nicotinaldehyde suggested the presence of either an essential lysine (Schiff's base formation) or cysteine residue (thiohemiacetal intermediate) at the YNDase active site. The relatively large value of the nicotinaldehyde inhibition constant (Ki = 68 microM), the observation that this analogue is a noncompetitive inhibitor of nicotinate formation, and the fact that this inhibition can be rendered irreversible through incubation with sodium borohydride, indicates that a Schiff's base intermediate is more likely to occur upon incubation of YNDase with nicotinaldehyde. However, YNDase is inactivated completely and irreversibly by N-ethylmaleimide at pH 6, and the enzyme is protected against this modification by either nicotinamide or nicotinate. These results suggest that both nicotinate and nicotinamide bind to YNDase, even though the enzymatic reaction is essentially irreversible, and that a cysteine residue may be present at the YNDase active site. PMID- 3036845 TI - Asymmetry of the Na+/H+ antiport of dog red cell ghosts. Sidedness of inhibition by amiloride. AB - Amiloride is a potent inhibitor of the Na+/H+ antiport. Inhibition is generally competitive with extracellular Na+ and therefore believed to result from binding to the outward-facing transport site. It is not known whether amiloride can interact with the internal aspect of the antiport. This question was addressed by trapping the drug inside resealed dog red cell ghosts. The antiport, which is quiescent in resting ghosts, was activated by acid-loading the cytoplasm. This was accomplished by exchanging extracellular Cl- for internal HCO-3 through capnophorin, the endogenous anion exchanger. The activity of the Na+/H+ antiport was detected as an increase in cell volume, resulting from the net osmotic gain associated with coupled Na+/H+ and Cl-/HCO-3 exchange, or as the uptake of 22Na+. Intracellular amiloride, at concentrations in excess of 100 microM, failed to inhibit Na+/H+ exchange. This is approximately 10 times higher than the concentration required for half-maximal inhibition when amiloride is added externally. Independent experiments demonstrated that failure of internal amiloride to inhibit exchange was not due to leakage of the inhibitor, to differences in pH, or to binding or inactivation of amiloride by the soluble contents. It was concluded that the antiport is functionally asymmetric with respect to amiloride. This implies that the transport site undergoes a conformational change upon translocation across the membrane or, alternatively, that a second site required for amiloride binding is only accessible from the outside. PMID- 3036846 TI - Characterization of the Na+/H+ exchanger during maturation of HL-60 cells induced by dimethyl sulfoxide. AB - We have studied the activity of the Na+/H+ exchanger during dimethyl sulfoxide (Me2SO)-induced maturation of the human promyelocytic leukemia cell line HL-60. 22Na uptake was measured in cells preloaded with Li+ or NH+4 in order to specifically activate the Na+/H+ exchanger. Measurement of the rate of uptake as a function of sodium concentration revealed a decrease in Km for Na+ from 38 +/- 3 to 13 +/- 1 mM after 20-24-h treatment with Me2SO. Vmax was not changed significantly. Inhibition of the exchanger by dimethylamiloride (DMA) and by acidic external pH was similar in treated and untreated cells. Thus it is unlikely that the Na+ binding site is altered. A change, however, was observed in the regulation of the exchanger by intracellular pH. In control cells maximal stimulation of the Na+ uptake was observed when the intracellular pH decreased from 7.25 to 7.00. In Me2SO-treated cells the 22Na uptake at intracellular pH 7.00 was greater than in the control and continued to increase as the intracellular pH was adjusted below 7.00, down to 6.75. This suggests that the Na+/H+ exchanger in Me2SO-treated cells is altered structurally in its allosteric H+ binding site. The appearance of this modified exchanger preceded by a period of days the appearance of a functional property characteristic of mature granulocytes, that is, the capability to produce superoxide, suggesting that the modified exchanger may be required for the expression of the mature phenotype. A second modification, a decrease in the Vmax of the 22Na uptake, occurred after 2 days treatment with Me2SO. This reduction may reflect a decrease in the number of functioning exchangers per cell. PMID- 3036847 TI - Purified RNA polymerase II recognizes specific termination sites during transcription in vitro. AB - We have studied the ability of certain well-defined prokaryotic DNA sequences to act as specific termination signals for highly purified calf thymus RNA polymerase II. We used duplex DNA fragments modified to direct efficient and specific transcription of defined DNA templates to follow transcription with RNA polymerase alone in the absence of additional protein factors. Elongation of RNA chains by RNA polymerase II is processive through most DNA sequences. However, certain DNA sequences serve as effective "intrinsic" terminators for RNA polymerase II; in this they resemble the "rho-independent" terminators for the bacterial RNA polymerase. Several rho-independent bacterial terminators are also able to act as termination signals for RNA polymerase II. However, there is no apparent correlation between the efficiency of termination for the bacterial enzyme and that found for the calf thymus enzyme. One very efficient bacterial terminator (phage T7 early terminator) gives no termination with RNA polymerase II, and we have identified at least two sites that cause the eukaryotic enzyme to terminate but have no effect on transcription by the bacterial enzyme. Hence, the signals recognized as intrinsic termination sites for the two enzymes are substantially different. All of the sites that act as intrinsic terminators for RNA polymerase II contain a series of consecutive thymidine residues in the nontranscribed DNA strand (T-run), and the 3' end of the completed RNA normally lies within this sequence. It is plausible that the T-run is part of the signal for an RNA polymerase II termination site; however, there is no apparent correlation between the number of T residues and the efficiency of the terminator, suggesting that other sequence elements are required for, or modulate, termination. Several lines of evidence suggest that the formation of RNA secondary structures in the nascent transcript is not an essential element of the intrinsic RNA polymerase II termination signal. PMID- 3036848 TI - Modification of the internal pH sensitivity of the Na+/H+ antiporter by parathyroid hormone in a cultured renal cell line. AB - Sodium-proton antiporter activity can be modulated through changes Vmax and/or intracellular proton sensitivity of the antiporter. To characterize a parathyroid hormone (PTH)-induced decrease in antiporter activity in a continuous renal cell line (opossum kidney cells), the extracellular sodium and intracellular proton dependence of amiloride-inhibitable 22Na uptake was studied. The Km for extracellular sodium at intracellular pH 6.32 was 28 mM and was unaltered by PTH, whereas the Vmax was decreased by 26%. When intracellular pH was set over the range 5.87-7.57 by the potassium-nigericin method, antiporter activity increased as intracellular pH decreased. Hill analysis revealed Hill coefficients of 1.25 and 1.01 and half-maximal antiporter activity at intracellular pH values of 6.90 and 6.35 for control and PTH-treated cells, respectively. PTH decreased the apparent Vmax at low pH by 15% and the intracellular pH at which Na+/H+ exchange is half-maximal by 0.55 pH units. PMID- 3036849 TI - EPR characterization of the iron-sulfur clusters in the NADH: ubiquinone oxidoreductase segment of the respiratory chain in Paracoccus denitrificans. AB - The physicochemical properties of the iron-sulfur clusters present in the NADH:ubiquinone oxidoreductase of Paracoccus denitrificans have been examined in the cytoplasmic membrane particles by redox potentiometry and EPR spectroscopy. Analogous to the iron-sulfur clusters present in the mitochondrial NADH: ubiquinone oxidoreductase, we have found two binuclear and three tetranuclear EPR detectable iron-sulfur clusters, namely, N-1a, N-1b, N-2, N-3, and N-4. In the bacterial system, the two binuclear clusters differ in line shape and in Em values; the cluster with more rhombic symmetry (gx,y,z = 1.918, 1.937, 2.029) has the Em7.0 value of -150 while the almost axial one (gx,y,z = 1.929, 1.941, 2.019) has Em7.0 of -270 mV. The Em of the former cluster is pH dependent (-60 mV/pH) as in the case of mammalian N-1a while the latter is pH independent as is the mammalian cluster N-1b. The pH-dependent P. denitrificans [2Fe-2S] cluster, which we have labeled N-1a, has an Em7.0 as high as that of N-2, in contrast to the mammalian N-1a. Thus N-1a is reducible with a physiological reductant, NADH in this bacterial system. The Em of the cluster N-2 is also pH dependent (Em7.0 = 130 mV) with a pK value near 7.7. The Em values of all other clusters exhibit no pH dependence as in the case of their mammalian counterparts. We have found that the cluster N-1a is the most labile component among the five iron-sulfur clusters and may give rise to variable relative spin concentrations and extremely low Em values due to the facile modifications of the microenvironment of the cluster. The P. denitrificans NADH:ubiquinone oxidoreductase provides a unique and useful site I model system where redox composition is similar to the mitochondrial enzyme but with fewer numbers of polypeptides (Yagi, T. (1986) Arch. Biochem. Biophys. 250, 302-311). PMID- 3036850 TI - Construction of a synthetic Holliday junction analog and characterization of its interaction with a Saccharomyces cerevisiae endonuclease that cleaves Holliday junctions. AB - We describe the construction and characterization of an oligonucleotide Holliday junction analog and characterize its interaction with a Saccharomyces cerevisiae endonuclease that cleaves Holliday junctions. A Holliday junction analog containing four duplex arms and 54 base pairs was constructed by annealing four unique synthetic oligonucleotides. Mixing curve analysis showed that the complex contained a 1:1:1:1 mol ratio of the four unique sequence strands. In addition, a linear duplex with a sequence identical to two of the junction arms was also constructed for use as a control fragment. High resolution gel exclusion chromatography was used to purify and characterize the synthetic junction. The synthetic Holliday junction was found to be a specific inhibitor of a S. cerevisiae enzyme that catalyzes the cleavage of Holliday junctions. Under standard cleavage conditions, 50% inhibition was observed at a synthetic Holliday junction to substrate ratio of 7/1, whereas no inhibition by linear duplex was observed at molar ratios in excess of 150/1. Kinetic analysis showed that Holliday junction was a competitive inhibitor of the reaction and had an apparent Ki = 2.5 nM, although the mode of inhibition was complex. The synthetic Holliday junction was not a substrate for the enzyme, but was found to form a specific complex with the enzyme as evidenced by polyacrylamide gel electrophoresis DNA binding assays. PMID- 3036851 TI - The distribution of Escherichia coli recA protein bound to duplex DNA with single stranded ends. AB - A short single-stranded tail on one end of an otherwise duplex DNA molecule enables recA protein, in the presence of ATP and MgCl2, to form a complex with the DNA which extends into the duplex portion of the molecule. Nuclease protection studies at a concentration of MgCl2 which permits homologous pairing showed that cleavage by restriction endonucleases at sites throughout the duplex region was inhibited, whereas digestion by DNase I was not affected. These results indicate that recA protein binds to the duplex portion of tailed DNA allowing access by DNase I to a random sample of the many sites at which it cleaves, but providing limited protection of the relatively rare restriction sites. Electron microscopy revealed that the recA nucleoprotein complex with duplex DNA is indeed a segmented or interrupted filament that, with time, extends further from the single-stranded tail into the duplex region. recA protein binding extended into the duplex region more rapidly for duplexes with 5' tails than for those with 3' tails. These observations show that recA protein translocates from a single-stranded region into duplex DNA in the form of a segmented filament by a mechanism that is not strongly polarized. PMID- 3036852 TI - Structure-function studies on bacteriorhodopsin. III. Total synthesis of a gene for bacterio-opsin and its expression in Escherichia coli. AB - We have chemically synthesized a DNA duplex of 757 base pairs which encodes the entire protein sequence of mature bacterio-opsin of Halobacterium halobium. The main aim of the synthesis was to facilitate site-specific mutagenesis in all parts of the gene by replacement of short restriction fragments by their counterparts containing the required nucleotide changes. Therefore, 30 unique restriction sites that are fairly evenly spaced were introduced in the synthetic DNA. A total of 28 oligonucleotides ranging in size from 21 to 69 nucleotides were synthesized corresponding to both strands. The entire gene was assembled from four synthetic fragments of 25, 268, 219, and 245 base pairs. The correctness of the nucleotide sequence was confirmed by sequencing the fragments as well as the complete gene. When expressed under the control of PL promoter in Escherichia coli, the synthetic and the native genes gave similar amounts of bacterio-opsin. Attempts to increase expression of the synthetic gene by introducing codons that are preferred in E. coli or by introduction of a synthetic transcription terminator were without significant effect. PMID- 3036853 TI - Vaccinia virus encapsidates a novel topoisomerase with the properties of a eucaryotic type I enzyme. AB - A DNA topoisomerase has been purified from vaccinia virus cores. The native enzyme is composed of a single subunit of 32,000 daltons. The enzyme relaxes both positively and negatively supercoiled DNA in the absence of an energy cofactor. Enzymatic activity is stimulated by magnesium ions and inhibited by ATP, and during relaxation the topoisomerase changes the linking number of the DNA substrate by steps of one. Trapping of the covalent DNA-enzyme intermediate has been achieved, and analysis of the cleavage of duplex DNA by the enzyme reveals that it makes a single-strand break and forms a covalent bond through the 3'-end of the broken strand. Enzymatic activity and formation of the trapped intermediate are inhibited by the drugs novobiocin, coumermycin A1, and berenil. The virally encapsidated enzyme has novel properties but most closely resembles a eucaryotic type I topoisomerase. PMID- 3036854 TI - The GTP-binding protein of rod outer segments. I. Role of each subunit in the GTP hydrolytic cycle. AB - The GTP-binding protein of Bufo marinus rod outer segments (ROS) is composed of 3 subunits: G alpha, 39,000; G beta, 36,000; and G gamma, approximately 6,500. A stepwise analysis of the GTP hydrolytic cycle (GTP binding, GTP hydrolysis, and GDP release) was facilitated by using purified subunits of the GTP-binding protein. When G alpha and G beta, gamma concentrations were held constant, the initial rate of guanosine-5'-O-(3-thiotriphosphate) (GTP gamma-s) binding to G alpha was dependent upon the amount of bleached rhodopsin present (as illuminated, urea-washed ROS disc membranes). When G alpha and the quantity of these membranes was held constant, the initial rate of GTP gamma-s binding to G alpha was markedly enhanced by increasing the amount of G beta, gamma. G beta preparations (free of G gamma) also stimulated the binding of GTP gamma-s to G alpha to the same extent as G beta, gamma preparations, suggesting that G gamma is not an essential component of the G beta, gamma-dependent stimulation of the rate of GTP gamma-s binding to G alpha. Nonlinear regression analysis revealed a single class of binding sites with an apparent stoichiometry of 1 mol of site/mol of G alpha under optimal binding conditions. Following GTP binding to G alpha, the GTP X G alpha complex dissociates from G beta, gamma which remains primarily bound to the ROS disc membranes. Moreover, while GTP remains in excess, the rates of GTP hydrolysis exhibited saturation in the presence of increasing amounts of G beta, gamma. Nonlinear regression analysis of these data argues against a direct role for G beta, gamma in the hydrolysis of GTP. Thus, both topologic and kinetic data support the concept that GTP hydrolysis is carried out by G alpha alone. After hydrolysis of GTP, the GDP X G alpha complex returned to the ROS disc membrane when G beta, gamma was present on the membrane surface, in the presence and absence of light. Without guanine nucleotides GDP release occurred in the presence of illuminated ROS disc membranes and G beta, gamma. Guanine nucleotides (GTP gamma-s approximately equal to GTP approximately equal to guanosine 5' (beta, gamma-imido)triphosphate greater than GDP) could effectively displace GDP from G alpha under these conditions.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3036855 TI - Purification and some properties of component B of the 4-chlorophenylacetate 3,4 dioxygenase from Pseudomonas species strain CBS 3. AB - 4-Chlorophenylacetate 3,4-dioxygenase system from Pseudomonas sp. CBS 3 consists of two protein components, a red-brown iron-sulfur protein (component A) which functions as dioxygenase and an orange-colored reductase (component B). Component B was purified by a five-step procedure. Criterion of purity was sodium dodecyl sulfate-polyacrylamide gel electrophoresis, which also showed that the protein consists of one polypeptide species with a molecular weight of 35,000. With gel chromatography on Sephadex G-100 also, a molecular weight of 35,000 was found for the native enzyme, suggesting a monomeric structure for the reductase enzyme. The isoelectric point was determined as pH 4.8. The visible absorption spectrum of the homogeneous protein exhibits maxima at 336, 394, and 458 nm. One mol of FMN, 2.1 mol of iron, and 1.7 mol of acid-labile sulfide were found in one mol of component B. The EPR-spectrum of the reduced form of the enzyme (with NADH) showed two types of signals. The signal at g values of 2.03, 1.94, and 1.90 was assigned to an iron-sulfur cluster of the [2Fe-2S]-type. The properties of the other signal type at g = 2.004 are characteristic of flavosemiquinone radical which does not show coupling to an other paramagnetic center. The apparent Km values for 2,6-dichlorophenolindophenol, cytochrome c, and NADH were calculated from Lineweaver-Burk plots as 6.3, 2.3, and 32 microM, respectively. Inhibitors of the reductase are metal-chelating reagents and sulfhydryl-inhibiting compounds. Component B reduces several redox compounds: 2,6 dichlorophenolindophenol, potassium hexacyanoferrat III, cytochrome c, methylene blue, and nitro blue tetrazolium. PMID- 3036856 TI - Properties of the exonuclease activity that degrades H4 histone mRNA. AB - We have described a cell-free system for studying mRNA degradation (Ross, J., and Kobs, G. (1986) J. Mol. Biol. 188, 579-593). Using that system we found that human H4 histone mRNA was degraded in a 3' to 5' direction by an exonucleolytic activity. Here we investigate several properties of the crude system and of the exonuclease. A RNase inhibitor, such as that from placenta, was required to block nonspecific ribonucleases and thereby to permit different mRNAs to be degraded at different rates. The histone mRNA exonuclease required divalent cation (magnesium) but not exogenously added ATP or GTP. It functioned efficiently at monovalent cation concentrations ranging from 0.5 to 200 mM. It was bound to ribosomes isolated from cells lysed in low salt buffers. However, it was eluted in active form from the ribosomes by exposing them to 0.3 M KCl. The enzyme rapidly degraded deproteinized, 32P-labeled histone mRNA prepared enzymatically. PMID- 3036857 TI - Substrate specificity of the exonuclease activity that degrades H4 histone mRNA. AB - We have investigated the substrate specificity of an exonuclease that degrades human H4 histone mRNA, using synthetic RNA templates incubated in a cell-free mRNA decay system (Ross, J., and Kobs, G. (1986) J. Mol. Biol. 188, 579-593). Five RNAs that lacked poly(A), including histone, were degraded rapidly in vitro. Polyadenylated histone mRNA was degraded at least 10-fold more slowly than unmodified histone mRNA. Double-stranded RNA and DNA were very stable. Single stranded DNA was degraded approximately 20-fold more slowly than single-stranded, non-polyadenylated RNA, and RNA with a 3' phosphoryl group was degraded more slowly than RNA with a 3'-hydroxyl group. Uncapped RNAs were degraded rapidly in the unfractionated system but were stable in reactions containing a ribosomal high salt wash extract. Therefore, the exonuclease activity released from ribosomes by high salt extraction was separated from the enzyme(s) that degraded uncapped RNAs. PMID- 3036858 TI - Reduction of collagen production in keloid fibroblast cultures by ethyl-3,4 dihydroxybenzoate. Inhibition of prolyl hydroxylase activity as a mechanism of action. AB - Excessive accumulation of collagen is the hallmark of several clinical conditions characterized by tissue fibrosis. Previously, 3,4-dihydroxybenzoic acid, a structural analog of alpha-ketoglutarate and ascorbate, has been shown to inhibit the activity of purified prolyl 4-hydroxylase, the enzyme catalyzing the synthesis of 4-hydroxyproline during intracellular biosynthesis of procollagen. In this study a hydrophobic modification, an ethyl ester, of 3,4-dihydroxybenzoic acid was tested for its effects on collagen synthesis and prolyl hydroxylase activity in human skin fibroblast cultures. The results indicated that 0.4 mM ethyl-3,4-dihydroxybenzoate markedly inhibited the synthesis of 4-hydroxyproline in normal cell cultures apparently as a result of reduced prolyl 4-hydroxylase activity, and the synthesis and secretion of both type I and type III procollagens were markedly reduced. Control experiments indicated that the test compound did not affect the viability, proliferation, or plating efficiency of the cells, and it had little, if any, effect on the synthesis of noncollagenous proteins. Furthermore, determinations of type I and type III procollagen mRNA steady-state levels by slot-blot hybridizations suggested that the inhibition of procollagen production did not occur on the pretranslational level. Thus, ethyl 3,4-dihydroxybenzoate selectively reduced procollagen production in fibroblast cultures by inhibiting the post-translational synthesis of 4-hydroxyproline. Similar inhibition was also observed in keloid fibroblast cultures, demonstrating the potential applicability of ethyl-3,4-dihydroxybenzoate, or other structural alpha-ketoglutarate or ascorbate analogs, for treatment of fibrotic diseases. PMID- 3036859 TI - Two nuclear factors compete for the skeletal muscle actin promoter. AB - We show that DNA fragments, which span the skeletal muscle alpha-actin promoter, form specific complexes with proteins from nuclear extracts of 11 different cultured cell types. These include two myocyte stage specific cell types, myoblast and myotube. The myocyte nuclear extracts are distinguished from all of the others by the electrophoretic mobility of the predominant DNA complex. This complex is greatly diminished in the nine non-myocyte cell types, where a different complex predominates. The major myocyte and non-myocyte complexes are caused by distinct binding activities that compete for the same DNA sequence element. DNA footprint analysis shows that this element is located 78 nucleotides upstream of the transcription start site, within a region that appears essential for expression in developing myocytes. PMID- 3036860 TI - Calcium-calmodulin stimulates inositol 1,4,5-trisphosphate kinase activity from insulin-secreting RINm5F cells. AB - In a cytosolic fraction derived from insulin-secreting RINm5F cells, the rate of conversion of inositol 1,4,5-trisphosphate (Ins-1,4,5-P3) to inositol 1,3,4,5 tetrakisphosphate (Ins-1,3,4,5-P4) was half-maximally stimulated by 0.8 microM Ca2+ (Biden, T. J., and Wollheim, C. B. (1986) J. Biol. Chem. 261, 11931-11934). In the present study we show that after initial purification by anion exchange chromatography, the Ins-1,4,5-P3 kinase activity responsible for that conversion is stimulated by Ca2+-calmodulin, but not by Ca2+ alone. This is almost certainly due to a specific interaction of the enzyme and its activator since kinase activity was retained on a calmodulin-linked Sepharose 6B column in the presence of Ca2+ but eluted upon chelation of the cation. After this two-step purification, Ins-1,4,5-P3 kinase activity was maximally stimulated 5-fold by 10 microM calmodulin in the presence of 10(-5) M Ca2+, and 2 1/2-fold at 10(-6) M Ca2+. Under these conditions the minimum concentrations of calmodulin needed to stimulate activity were in the 10-50 nM range. At 10(-7) M Ca2+, calmodulin (up to 30 microM) was without effect. Stimulated Ins-1,4,5-P3 kinase activity was inhibited in a dose-dependent fashion by N-(6-aminohexyl)-5-chloro-1 naphthalenesulfonamide (W7) although the calmodulin antagonist had no effect on the residual activity seen at 10(-7) M Ca2+. These results strongly support our previous suggestion that alterations in cytosolic free Ca2+ concentrations play an important role in regulating the levels of Ins-1,4,5-P3 and Ins-1,3,4,5-P4 during cellular stimulation. PMID- 3036861 TI - The metabolism of inositol 1,3,4-trisphosphate to inositol 1,3-bisphosphate. AB - We previously demonstrated a pathway for the metabolism of inositol 1,3,4 trisphosphate (Ins(1,3,4)P3) to inositol 3,4-bisphosphate (Ins(3,4)P2) in calf brain extracts. Inositol polyphosphate 1-phosphatase, a Mg2+-dependent, lithium ion-inhibited enzyme, specifically hydrolyzes Ins(1,3,4)P3 to Ins(3,4)P2 and Ins(1,4)P2 to Ins 4-P (Inhorn, R. C., Bansal, V. S., and Majerus, P. W. (1987) Proc. Natl. Acad. Sci. U.S.A. 84, 2170-2174). Now we have found an alternative pathway for the metabolism of Ins(1,3,4)P3 in crude calf brain extracts. Along this pathway, Ins(1,3,4)P3 is first converted to Ins(1,3)P2 which is further hydrolyzed to Ins 1-P. This pathway involves a 4-phosphatase and a 3-phosphatase which do not require Mg2+ and are not inhibited by lithium ions. A similar 4 phosphatase also degrades Ins(3,4)P2 to Ins 3-P. Three different inositol bisphosphates formed from calf brain supernatant are each further metabolized by a separate enzyme. The three inositol monophosphates, i.e. Ins 1-P, Ins 3-P, and Ins 4-P, are converted to inositol by inositol monophosphate phosphatase (Ackermann, K. E., Gish, B. G., Honchar, M. P., and Sherman, W. R. (1987) Biochem. J. 242, 517-524). PMID- 3036862 TI - A tyrosine kinase related to pp60c-src is associated with membranes of Electrophorus electricus electric organ. AB - A tyrosine-specific protein kinase immunologically related to pp60c-src, the cellular homolog of the Rous sarcoma virus-transforming protein, was expressed at elevated levels in the electric organ of the electric eel Electrophorus electricus. The electric organ kinase phosphorylated antibodies reactive with pp60c-src at tyrosine residues in immune complex protein kinase assays and was associated with electric organ membranes enriched in acetylcholine receptors. The protein recognized by anti-pp60c-src antibodies was phosphorylated in endogenous membrane phosphorylation reactions and was shown to have a relative molecular mass of 57 kDa by two-dimensional gel electrophoresis. In immune complex protein kinase assays the 57-kDa protein was phosphorylated at threonine by a distinct threonine kinase from the electric organ. The tyrosine kinase was purified 844 fold from electric organ membranes by chromatography on omega-aminohexyl agarose, phosphocellulose, and casein-Sepharose. Threonine kinase activity in immunoprecipitates was not observed in the tyrosine kinase fractions after the first step. Incubation of the casein Sepharose fraction with [gamma-32P]ATP-Mn2+ in solution resulted in phosphorylation of only the 57-kDa protein. Phosphorylation occurred solely at tyrosine, suggesting that the kinase is capable of autophosphorylation. The structural and functional properties of the 57-kDa electric organ kinase indicate that the 57-kDa electric organ protein is a member of the src subfamily of tyrosine kinases and is closely related to pp60c src. PMID- 3036863 TI - Regulation of thyrotropin-releasing hormone receptor binding and phospholipase C activation by a single GTP-binding protein. AB - In a crude membrane preparation of rat 7315c cells, GTP was found to enhance thyrotropin-releasing hormone- (TRH) stimulated inositol triphosphate (IP3) formation with a potency of 0.97 +/- 0.1 microM. TRH stimulation of IP3 formation was inhibited by high GDP concentrations. Neither nucleotide had any effect in the absence of TRH. 5'-Guanosine gamma-thiotriphosphate (GTP gamma S) stimulated IP3 formation in the absence of TRH; the apparent affinity of GTP gamma S was 0.16 +/- 0.05 microM. GTP blocked GTP gamma S stimulation of IP3 formation in a concentration-dependent manner. The apparent affinity of GTP for the site of action shared by GTP gamma S was calculated to be 0.98 +/- 0.3 microM. TRH was able to reverse inhibition of GTP gamma S-stimulated IP3 formation by GTP but could not reverse inhibition by GDP. A lag in the rate of IP3 formation in response to GTP gamma S was abolished by addition of TRH. These data support the proposal that activation of the TRH receptor enhances turnover of guanine nucleotides at the binding protein coupling the receptor to phospholipase C. In addition, GTP gamma S diminished high affinity [3H]Me-TRH binding. The potency of GTP gamma S at decreasing [3H]Me-TRH binding was 0.092 +/- 0.03 microM. GTP gamma S (0.1 microM) decreased the affinity of the TRH receptor for [3H]Me-TRH from 2 to 100 nM. Maximally effective concentrations of GTP gamma S, Gpp(NH)p, GTP, and GDP decreased specific [3H]Me-TRH binding by 80%. Pretreatment of cells with pertussis toxin (30 ng/ml for 24 h) failed to affect TRH receptor affinity or the potency or efficacy of GTP gamma S in diminishing [3H]Me-TRH binding, supporting the identification of Gp (a GTP-binding protein associated with phospholipase C and Ca2+-mobilizing receptors) as distinct from Gi (an inhibitory GTP-binding protein). In contrast to its lack of effect on TRH receptor binding, 3-h pertussis toxin treatment decreased agonist affinity of the mu-opiate receptor and abolished the ability of GTP gamma S to shift the affinity of the mu-opiate receptor for its agonist. The affinities calculated for GTP, GDP, GTP gamma S, and Gpp (NH)p for the G-protein regulating receptor affinity and IP3 formation are nearly identical for each guanine nucleotide tested, suggesting the same G protein regulates both activities. PMID- 3036864 TI - Yeast cytochrome c peroxidase. Coordination and spin states of heme prosthetic group. AB - Electronic absorption and electron paramagnetic resonance (EPR) spectroscopic examinations revealed that a freshly prepared cytochrome c peroxidase (CCP) contains a penta-coordinated high spin ferric protoheme group. The penta coordinated high spin state of fresh CCP is maintained in a remarkably wide range of pH (4-8). The freezing of fresh CCP induces the reversible coordination of an internal strong field ligand to the heme iron to form a hexa-coordinated low spin compound, which shows EPR extrema at gx = 2.70, gy = 2.20 and gz = 1.78. In the presence of glycerol the freezing-induced artifacts are eliminated and the fresh enzyme exhibits an EPR spectrum of rhombically distorted axial symmetry with EPR extrema at gx = 6.4, gy = 5.3, and gz = 1.97 at 10 K, characteristic of the penta coordinated high spin enzyme. Upon aging CCP is converted to a hexa-coordinated high spin state due to the coordination of an internal weak field ligand to the heme iron. This conversion is accelerated at acidic pH values, and its reversibility varies from fully reversible to irreversible depending on the degree of enzyme aging. The aging-induced hexa-coordinated CCP is unreactive with hydrogen peroxide and exhibits an EPR spectrum of purely axial symmetry with extrema at g = 6 and g = 2 and an electronic absorption spectrum with an intensified Soret band at 408 nm (epsilon 408 nm = 120 mM-1 cm-1) and a blue shifted charge-transfer band at 620 nm. Spectroscopic properties of different coordination and spin states of fresh and aged CCPs are compiled in order to formulate a generalized spectroscopic characterization of penta- and hexa coordinated high spin ferric hemoproteins. PMID- 3036865 TI - Mechanism of inhibition of glycolysis by vanadate. AB - Vanadate is known to inhibit several phosphatases including Na+, K+-ATPase, alkaline phosphatase, and glyceraldehyde-3-P dehydrogenase. Inhibition presumably results because vanadium adopts a stable structure which resembles the transition state of phosphate during the reactions involving these enzymes. We performed experiments to further examine the effects of vanadate (VO3-4) on erythrocyte (red blood cells (RBC] glycolytic intermediates. RBC obtained from human subjects were centrifuged and washed with lactated Ringer's 5% dextrose. 31P nuclear magnetic resonance analysis of the RBC revealed the characteristic peaks for the 3-phosphate and 2-phosphate of 2,3-diphosphoglycerate (DPG), inorganic phosphate (Pi), and ATP. Incubation of RBC with 10(-6) M VO3-4 led to a disappearance of ATP and 2,3-DPG while the peak for Pi increased. By the end of 4 h over 90% of the VO3-4 had been reduced to VO2+ (vanadyl) in the RBC. The effects of 10(-4) M iodoacetamide and 10(-5) M ethacrynic acid, known inhibitors of glyceraldehyde-3 P dehydrogenase that act by interactions with sulfhydryl groups (-SH) of the enzyme, were similar to those of VO3-4. Incubation with vanadyl did not affect the peaks for Pi, 2-DPG, or 3-DPG. Furthermore, using electron spin resonance we demonstrated that in the presence of glyceraldehyde-3-P dehydrogenase, VO3-4 is reduced to VO2+. The findings demonstrate that VO3-4 inhibits glycolysis at micromolar concentrations and that the ion is reduced to VO2+ in the cell. The similarity of the effect of VO3-4 to those of iodoacetamide and ethacrynic acid suggests that interactions with -SH groups is its mechanism of inhibition. Since under physiological conditions intracellular VO3-4 concentrations are in the micromolar range and may exist in oxidized and/or reduced forms, VO3-4 could regulate the activity of glyceraldehyde-3-P dehydrogenase through changes in the redox state of the enzyme rather than by substituting for the PO3-4 ion. PMID- 3036866 TI - Phosphorylation of the 20,000-dalton light chain of smooth muscle myosin by the calcium-activated, phospholipid-dependent protein kinase. Phosphorylation sites and effects of phosphorylation. AB - Smooth muscle heavy meromyosin (HMM) is phosphorylated by the Ca2+-activated phospholipid-dependent protein kinase, i.e. protein kinase C, at three sites on each 20,000-dalton light chain. Phosphorylation of three sites also is observed with isolated 20,000-dalton light chain and HMM subfragment 1. The phosphorylation sites are serine 1, serine 2, and threonine 9. Threonine is phosphorylated most rapidly followed by either serine 1 or 2. Phosphorylation of the third site occurs only on prolonged incubation. Phosphorylation is a random process. HMM phosphorylated at two sites per light chain by protein kinase C can be dephosphorylated, as shown using two phosphatase preparations. Increasing levels of phosphorylation of HMM by protein kinase C causes a progressive inhibition of the subsequent rate of phosphorylation of serine 19 by myosin light chain kinase and causes a progressive inhibition of actin-activated ATPase activity of HMM, prephosphorylated by myosin light chain kinase. Inhibition of ATPase activity is due to a decreased affinity of HMM for actin rather than a change in Vmax. Previous results with HMM and protein kinase C (Nishikawa, M., Sellers, J. R., Adelstein, R. S., and Hidaka, H. (1984) J. Biol. Chem. 259, 8808 8814) examined effects induced by phosphorylation of the threonine residues. Our results confirm these and consider also the influence of higher levels of phosphorylation by protein kinase C. PMID- 3036867 TI - Primary structure of the human 4F2 antigen heavy chain predicts a transmembrane protein with a cytoplasmic NH2 terminus. AB - The human cell surface antigen 4F2 is a disulfide-linked heterodimer consisting of a glycosylated heavy chain and a nonglycosylated light chain. The antigen is ubiquitously expressed on proliferating cells but only in resting cells from certain tissues. Its function has been proposed to relate to cellular Ca2+/Na+ exchange. We describe the molecular cloning of the 4F2 heavy chain gene and cDNA by a gene transfer approach. Part of the gene was isolated from a genomic lambda library constructed with DNA of a secondary transfectant L cell line that expresses 4F2 antigen. A gene-specific probe derived from the phage inserts was used to isolate two full length cDNA clones. Both cDNA clones directed the expression of 4F2 antigen in transfected mouse L cells. The 4F2 antigen heavy chain gene specifies a 2.1-kilobase mRNA with an open reading frame coding for a 529-residue protein of 58 kDa. The protein lacks an NH2-terminal signal peptide but contains an internal transmembrane-spanning region and four potential glycosylation sites in its COOH-terminal domain. We predict that the 4F2 antigen heavy chain is a transmembrane protein with a cytoplasmic NH2 terminus of 81 amino acids. The antigen shows no homology to known protein sequences. PMID- 3036868 TI - The Na+/H+ exchanger is constitutively activated in P19 embryonal carcinoma cells, but not in a differentiated derivative. Responsiveness to growth factors and other stimuli. AB - We have examined the functional properties and growth factor responsiveness of the plasma membrane Na+/H+ exchanger in pluripotent P19 embryonal carcinoma (EC) cells and in a differentiated mesodermal derivative (MES-1) by analyzing the recovery of cytoplasmic pH (pHi) from an acute acid load under bicarbonate-free conditions. In the absence of exogenous growth factors, the mean steady-state pHi of undifferentiated P19 cells (7.49 +/- 0.03) is 0.55 unit higher than the value of differentiated MES-1 cells (6.94 +/- 0.01). In both cell types, recovery of pHi from an NH+4-induced acid load follows an exponential time course and is entirely mediated by the amiloride-sensitive Na+/H+ exchanger in the plasma membrane. Kinetic analysis indicates that the higher steady-state pHi in P19 EC cells is due to an alkaline shift in the pHi sensitivity of the Na+/H+ exchange rate, as compared to that in MES-1 cells. The Na+/H+ exchanger of MES-1 cells is responsive to epidermal growth factor, platelet-derived growth factor, serum, phorbol esters, and diacylglycerol, as shown by a rapid amiloride-sensitive rise in pHi of 0.15-0.35 unit. This mitogen-induced alkalinization is attributable to an alteration in the pHi sensitivity of the exchanger. In contrast, the Na+/H+ exchanger of P19 EC cells fails to respond to any of these stimuli. Similarly, hypertonic medium rapidly activates the Na+/H+ exchanger in MES-1, but not in P19 EC cells. We conclude that the Na+/H+ exchanger in undifferentiated P19 EC stem cells is maintained in a fully activated state which is unaffected by extracellular stimuli, as if signal pathways normally involved in growth factor action are constitutively operative. PMID- 3036870 TI - The primary structure of a procaryotic glycoprotein. Cloning and sequencing of the cell surface glycoprotein gene of halobacteria. AB - The hexagonally patterned surface layer of halobacteria consists of a true glycoprotein. This procaryotic glycoprotein has recently been shown to exhibit novel features with respect to saccharide structure and saccharide biosynthesis. The primary structure and the location of glycosylation sites were determined by cloning and sequencing of the glycoprotein gene of Halobacterium halobium. According to the predicted amino acid sequence, the glycoprotein is synthesized with a N-terminal leader sequence of 34 amino acid residues reminiscent of eucaryotic and procaryotic signal peptides. A hydrophobic stretch of 21 amino acid residues at the C terminus probably serves as a transmembrane domain. 14 threonine residues are clustered adjacent to this membrane anchor and linked to these threonines are all the disaccharides of the cell surface glycoprotein. 12 N glycosylation sites are distributed over the polypeptide chain. PMID- 3036869 TI - Analysis of the tissue-specific expression, developmental regulation, and linkage relationships of a rodent gene encoding heart fatty acid binding protein. AB - The rat contains at least three homologous cytosolic proteins that bind long chain fatty acids, termed liver (L-), intestinal (I-), and heart (H-) fatty acid binding protein (FABP). I-FABP mRNA is confined to the gastrointestinal tract while L-FABP mRNA is abundantly represented in hepatocytes as well as enterocytes. We have isolated a rat heart FABP cDNA clone and determined the pattern of H-FABP mRNA accumulation in a wide variety of tissues harvested from late fetal, suckling, weaning, and adult rats. RNA blot hybridizations and primer extension analysis disclosed that the distribution of H-FABP mRNA in adult rat tissues is different from that of I- or L-FABP mRNA. H-FABP mRNA is most abundant in adult heart. This mRNA was also present in an adult slow twitch (type I) skeletal muscle (soleus, 63% of the concentration in heart), testes (28%), a fast twitch skeletal muscle (psoas, 17%), brain (10%), kidney (5%), and adrenal gland (5%). H-FABP mRNA was not detected in adult small intestine, colon, spleen, lung, or liver RNA. Distinct patterns of developmental change in H-FABP mRNA accumulation were documented in heart, placenta, brain, kidney, and testes. Myocardial H-FABP mRNA levels rise rapidly during the 48 h prior to and after birth, reaching peak levels by the early weaning period. The postnatal increase in myocardial H-FABP mRNA concentration and its relative distribution in adult fast and slow twitch skeletal muscle are consistent with its previously proposed function in facilitating mitochondrial beta-oxidation of fatty acids. However, the presence of H-FABP mRNA in brain, a tissue which does not normally significantly oxidize fatty acids in late postnatal life, suggests that H-FABP may play a wider role in fatty acid metabolism than previously realized. Mouse hamster somatic cell hybrids were utilized to map H-FABP. Using stringencies which did not produce cross-hybridization between L-, I-, and H-FABP DNA sequences, we found at least three loci in the mouse genome, each located on different chromosomes, which reacted with our cloned H-FABP cDNA. None of these H FABP-related loci were linked to the gene which specifies a highly homologous adipocyte-specific protein termed aP2 or to genes encoding two other members of this protein family, cellular retinol binding protein and cellular retinol binding protein II.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3036872 TI - Modulation of the cellular ratio of chromosomal high mobility group proteins 14 to 17 in transfected cells. AB - The cDNAs coding for human nonhistone chromosomal high mobility group (HMG) proteins 14 and 17 have been introduced into the eukaryotic expression vector pSVL under the transcriptional control of the SV40 late promoter and the constructs used to transfect COS cells. Transfection with plasmid pSVL14s, containing the HMG-14 cDNA in the sense orientation, increased the endogenous levels of HMG-14 mRNA 50-fold and the levels of HMG-14 protein 3-fold. Transfection with pSVL17s, which contains the HMG-17 mRNA in the sense orientation, resulted in a 19-fold increase in mRNA levels and a 3-fold increase in the protein level. Transfection with pSVL17as, containing the HMG-17 in the antisense orientation, resulted in a noticeable decrease in the protein levels. The overproduction of HMG mRNAs does not affect the level of other cellular mRNAs and the increase in the cellular level of either HMG-14 or -17 did not affect the level of the other HMG or that of any other cellular protein examined. The results suggest that COS cells can tolerate large excess of HMG mRNAs and protein, that the relative amounts of HMG-14 and HMG-17 and their mRNAs are not constant, and that neither the transcription nor the translation of the proteins is coordinately regulated. PMID- 3036871 TI - Biosynthetic and glycosylation studies of cell surface platelet-derived growth factor receptors. AB - The cell surface pool of metabolically labeled platelet-derived growth factor (PDGF) receptors in BALB/c3T3 fibroblasts was studied using an antiphosphotyrosine antibody. Exposure of intact cells to PDGF stimulates autophosphorylation of surface PDGF receptors and allowed immunoaffinity purification of only PDGF-activated receptors. Pulse-chase experiments demonstrated appearance of newly synthesized receptors in a surface activatable pool within 30-45 min of synthesis. In the absence of exogenous PDGF, the apparent half-life of this pool was 2 h. The presence of both N- and O-linked oligosaccharide chains on cell surface PDGF receptors was demonstrated. Enzymatic removal of the N-linked oligosaccharide chains reduced the receptor's apparent Mr by approximately 40 kDa and removal of O-linked oligosaccharide caused approximately a 7-kDa reduction. Activation of receptor tyrosine autophosphorylation by PDGF did not require either processing of high-mannose N linked oligosaccharides to complex forms or the presence of sialic acid on receptor oligosaccharide chains. Tryptic cleavage of PDGF-activated surface receptors in intact cells yielded two discrete phosphotyrosine-containing fragments of 107 and 85 kDa. Cleveland digest patterns from each fragment indicate that both are derived from the intact PDGF receptor. These data indicate that PDGF receptors are synthesized and turn over rapidly in the absence of ligand. Partial characterization of the extracellular domain oligosaccharide contribution to receptor function and trypsin susceptibility is provided. PMID- 3036873 TI - Role of the aggregation factor in the regulation of phosphoinositide metabolism in sponges. Possible consequences on calcium efflux and on mitogenesis. AB - The aggregation factor (AF) of the marine sponge Geodia cydonium recognizes the aggregation receptor (AR) which is inserted in the plasma membrane, under formation of species-specific aggregates. The specific cell-binding fragment of the AF was used to investigate for the first time the phosphoinositide metabolism in a lower avertebrate system. We found that after binding of the cell-binding fragment to the aggregation receptor a strong and rapid stimulation of the phosphate incorporation into phosphatidylinositol occurs followed by an increased turnover of phosphoinositides in the Geodia cells. The consequences of an increased degradation of phosphatidylinositol 4,5-bisphosphate into the two second messengers inositol-1,4,5-trisphosphate and diacylglycerol are 2-fold. First, after the addition of the extracellular stimulus the cytosolic Ca2+ concentration rises, resulting in a rapid increased Ca2+ efflux rate. The functional consequence of the increase of the extracellular Ca2+ level is an initiation of the aggregate formation that is mediated by the collagen assembly factor (= primary aggregation factor). Second, some experimental evidences are presented, showing that the other second messenger formed, diacylglycerol, causes a translocation of protein kinase C within the cell. Incubation of Geodia cells with the cell-binding fragment of the AF, or with the phorbol ester, 12-O tetradecanoylphorbol-13-acetate, resulted within 5 min after treatment in a 70% decrease in protein kinase C activity in the cytosolic fraction and in a 700% increase in enzyme activity in the membrane fraction. It is proposed that by membrane association protein kinase C becomes activated. As a result of this event a series of cellular proteins are phosphorylated, a process which ultimately leads to an unusually strong induction of DNA polymerase alpha activity. From these data we conclude that inositol trisphosphate and protein kinase C also play a fundamental role in cellular signal transduction in lower eukaryotes. PMID- 3036875 TI - Protein damage and degradation by oxygen radicals. I. general aspects. AB - Aggregation, fragmentation, amino acid modification, and proteolytic susceptibility have been studied following exposure of 17 proteins to oxygen radicals. The hydroxyl radical (.OH) produced covalently bound protein aggregates, but few or no fragmentation products. Extensive changes in net electrical charge (both + and -) were observed. Tryptophan was rapidly lost with .OH exposure, and significant production of bityrosine biphenol occurred. When incubated with cell-free extracts of human and rabbit erythrocytes, rabbit reticulocytes, or Escherichia coli, most .OH-modified proteins were proteolytically degraded up to 50 times faster than untreated proteins. The exceptions were alpha-casein and globin, which were rapidly degraded without .OH modification. ATP did not stimulate the degradation of .OH-modified proteins, but alpha-casein was more rapidly degraded. Leupeptin had little effect under any condition, and degradation was maximal at pH 7.8. The data indicate that proteins which have been denatured by .OH can be recognized and degraded rapidly and selectively by intracellular proteolytic systems. In both red blood cells and E. coli, the degradation appears to be conducted by soluble, ATP-independent (nonlysosomal) proteolytic enzymes. In contrast with the above results, superoxide (O2-) did not cause aggregation or fragmentation, tryptophan loss, or bityrosine production. The combination of .OH + O2- (+O2), which may mimic biological exposure to oxygen radicals, induced charge changes, tryptophan loss, and bityrosine production. The pattern of such changes was similar to that seen with .OH alone, although the extent was generally less severe. In contrast with .OH alone, however, .OH + O2- (+O2) caused extensive protein fragmentation and little or no aggregation. More than 98% of the protein fragments had molecular weights greater than 5000 and formed clusters of ionic and hydrophobic bonds which could be dispersed by denaturing agents. The results indicate a general sensitivity of proteins to oxygen radicals. Oxidative modification can involve direct fragmentation or may provide denatured substrates for intracellular proteolysis. PMID- 3036874 TI - A protein phosphatase associated with rat heavy gastric membranes enriched with (H+-K+)-ATPase influences membrane K+ transport activity. AB - Rat stimulated heavy gastric membranes enriched with (H+-K+)-ATPase, a marker for the apical membrane of the parietal cell, displayed a 32P-histone dephosphorylating activity which appeared to be physically copurified with, but functionally independent of, the ATPase. The protein phosphatase activity was optimal at pH 7.5 and was inhibited by fluoride (50 mM), inorganic phosphate (50 mM), and p-chloromercuribenzoate (0.1 mM), but was insensitive to vanadate (1 mM). The 32P-phosphoproteins in the heavy gastric membranes were also dephosphorylated, apparently by their own membrane-bound phosphatase in the presence of Mg2+ at millimolar concentrations, which is likely to enhance membrane-membrane interaction. Heavy gastric membrane vesicles incubated with Mg2+ (2 mM) exhibited no alterations in K+-dependent ATP-hydrolyzing activity, Cl permeability, and protein and lipid compositions, but irreversibly lost the ATP, K+-dependent H+-pumping activity. Since valinomycin, a K+-specific ionophore, restored the intravesicular acidifying activity and an inhibitor of the protein phosphatase, inorganic phosphate, largely blocked the Mg2+-induced change in the membrane transport function, it is reasonable to propose that the phosphatase action on certain membrane proteins, possibly the putative K+ transporter or regulatory proteins, selectively decreases K+-conductance in the apical membranes of gastric parietal cells. PMID- 3036876 TI - Protein damage and degradation by oxygen radicals. II. Modification of amino acids. AB - Exposure of proteins to the hydroxyl radical (.OH) or to the combination of .OH plus the superoxide anion radical (.OH + O2-) causes gross structural modification. Such modified proteins can undergo spontaneous fragmentation or can exhibit substantial increases in proteolytic susceptibility. In the present study, with the representative protein bovine serum albumin (BSA), we report that alterations to primary structure underlie such gross structural modifications. All amino acids in BSA were susceptible to modification by both .OH and .OH + O2- +O2), although tryptophan, tyrosine, histidine, and cysteine were particularly sensitive. At a radical/BSA molar ratio (nmol of radicals/nmol of BSA) of 10, we observed an average 9-10% destruction of amino acids; whereas at a ratio of 100, the average loss was 45%. Decreasing tryptophan fluorescence provided a useful index of amino acid loss and exhibited a clear dose dependence with .OH or with .OH + O2- (+O2). Linear production of the biphenol bityrosine was observed with .OH treatment. In contrast, .OH + O2- (+O2) induced only a limited bityrosine production rate which reached an early plateau. Studies with various chemical scavengers (t-butyl alcohol, isopropyl alcohol, mannitol, urate) and gasses (N2O, N2, O2, air) revealed that .OH is the primary radical responsible for all amino acid modifications, but that O2- and O2 can further transform the products of .OH reactions. Thus, O2-/O2 can potentiate .OH-dependent destruction of many amino acids (e.g. tryptophan) while inhibiting production of bityrosine by reacting with tyrosyl (phenoxyl) radicals. No amino acid loss or bityrosine production occurred with exposure to O2- (+O2) alone. Amino acid modifications caused both by .OH alone and by .OH + O2- (+O2) progressively affected the overall electrical charge of BSA. In a pH range of 3.7-6.2, some 16 new isoelectric focusing bands were induced by .OH, and some eight new bands were induced by .OH + O2- (+O2). The alterations to primary structure observed provide the key to an understanding of the link between oxidative modification and increased proteolytic susceptibility. PMID- 3036878 TI - Protein damage and degradation by oxygen radicals. IV. Degradation of denatured protein. AB - Proteolytic degradation of oxidatively damaged [3H] bovine serum albumin [( 3H]BSA) was studied during incubation with cell-free erythrocyte extracts and a wide variety (14) of purified proteases. [3H]BSA was pretreated by exposure (60Co radiation) to the hydroxyl radical (.OH), the superoxide anion radical (O2-), or the combination of .OH + O2- + oxygen. Treated (and untreated) samples were dialyzed and then incubated with erythrocyte extract or proteases for measurements of proteolytic susceptibility (release of acid-soluble counts). Both .OH and .OH + O2- + caused severalfold increases in proteolytic susceptibility (with extract and proteases), but O2- alone had no effect. Proteolytic susceptibility reached a maximum at 15 nmol of .OH/nmol of BSA and declined thereafter. In contrast, proteolytic susceptibility was still increasing at an .OH + O2-/BSA molar ratio of 100 (50% .OH + 50% O2-). Degradation in erythrocyte extracts was conducted by a novel ATP- and Ca2+-independent pathway, with maximal activity at pH 7.8. Inhibitor profiles indicate that this pathway may involve metalloproteases and serine proteases. Comparisons of proteolytic susceptibility with multiple modifications to BSA primary, secondary, and tertiary structure revealed a high correlation (r = 0.98) with denaturation/increased hydrophobicity by low concentrations of .OH. Covalent aggregation reactions (BSA cross-linking) may explain the declining proteolytic susceptibility observed at .OH/BSA molar ratios greater than 20. Protein denaturation may also have caused the increased proteolytic susceptibility induced by .OH + O2- + O2, but no simple correlation could be obtained. Results with .OH + O2- + O2 appear to have been complicated by direct BSA fragmentation reactions involving (.OH-induced) protein radicals and oxygen. These data indicate a direct and quantitative relationship between protein damage by oxygen radicals and increased proteolytic susceptibility. Oxidative denaturation may exemplify a simple, yet effective inherent mechanism for intracellular proteolysis. PMID- 3036877 TI - Protein damage and degradation by oxygen radicals. III. Modification of secondary and tertiary structure. AB - Proteins which have been exposed to the hydroxyl radical (.OH) or to the combination of .OH plus the superoxide anion radical and oxygen (.OH + O2- + O2) exhibit altered primary structure and increased proteolytic susceptibility. The present work reveals that alterations to primary structure result in gross distortions of secondary and tertiary structure. Denaturation/increased hydrophobicity of bovine serum albumin (BSA) by .OH, or by .OH + O2- + O2 was maximal at a radical/BSA molar ratio of 24 (all .OH or 50% .OH + 50% O2-). BSA exposed to .OH also underwent progressive covalent cross-linking to form dimers, trimers, and tetramers, partially due to the formation of intermolecular bityrosine. In contrast, .OH + O2- + O2 caused spontaneous BSA fragmentation. Fragmentation of BSA produced new carbonyl groups with no apparent increase in free amino groups. Fragmentation may involve reaction of (.OH-induced) alpha carbon radicals with O2 to form peroxyl radicals which decompose to fragment the polypeptide chain at the alpha-carbon, rather than at peptide bonds. BSA fragments induced by .OH + O2- + O2 exhibited molecular weights of 7,000-60,000 following electrophoresis under denaturing conditions, but could be visualized as hydrophobic aggregates in nondenaturing gels (confirmed with [3H]BSA following treatment with urea or acid). Combinations of various chemical radical scavengers (mannitol, urate, t-butyl alcohol, isopropyl alcohol) and gases (N2O, O2, N2) revealed that .OH is the primary species responsible for alteration of BSA secondary and tertiary structure. Oxygen, and O2- serve only to modify the outcome of .OH reaction. Furthermore, direct studies of O2- + O2 (in the absence of .OH) revealed no measurable changes in BSA structure. The process of denaturation/increased hydrophobicity was found to precede either covalent cross linking (by .OH) or fragmentation (by .OH + O2- + O2). Denaturation was half maximal at a radical/BSA molar ratio of 9.6, whereas half-maximal aggregation or fragmentation occurred at a ratio of 19.4. Denaturation/hydrophobicity may hold important clues for the mechanism(s) by which oxygen radicals can increase proteolytic susceptibility. PMID- 3036879 TI - Sources of variability in foot and mouth disease vaccine potency estimates based on serum neutralizing antibody assay. AB - Some vaccines can be assayed for potency by measuring the serum antibody response they produce in vaccinated test animals. Using data obtained from potency assays on batches of foot and mouth disease vaccine, the sources of variability in such a method were examined. A linear model is proposed for the analysis of replicate serum neutralizing antibody assays, which represents an improvement on the usual approach of working with only a mean serum assay value for each test animal. Components of variance were calculated, allowing the relative importance of the numbers of test animals, or the numbers of serum assays per test animal, to be estimated in terms of the variability of the overall group mean antibody response. A method is described for calculating fiducial intervals for the serological potency estimates, and it is shown that these intervals are no larger than, and are in fact probably smaller than, those obtained from quantal challenge tests. The results have important implications for the design and analysis of similar biological tests used for other products. PMID- 3036881 TI - The autodestructive consequences of thermal injury. PMID- 3036880 TI - Chemically modified collagen: a natural biomaterial for tissue replacement. AB - Glutaraldehyde crosslinking of native or reconstituted collagen fibrils and tissues rich in collagen significantly reduces biodegradation. Other aldehydes are less efficient than glutaraldehyde in generating chemically, biologically, and thermally stable crosslinks. Tissues crosslinked with glutaraldehyde retain many of the viscoelastic properties of the native collagen fibrillar network which render them suitable for bioprostheses. Implants of collagenous materials crosslinked with glutaraldehyde are subject long-term to calcification, biodegradation, and low-grade immune reactions. We have attempted to overcome these problems by enhancing crosslinking through bridging of activated carboxyl groups with diamines and using glutaraldehyde to crosslink the epsilon-NH2 groups in collagen and the unreacted amines introduced by aliphatic diamines. This crosslinking reduces tissue degradation and nearly eliminates humoral antibody induction. Covalent binding of diphosphonates, specifically 3-amino-1 hydroxypropane-1, 1-diphosphonic acid (3-APD), and chondroitin sulfate to collagen or to the crosslink-enhanced collagen network reduces its potential for calcification. Platelet aggregation is also reduced by glutaraldehyde crosslinking and nearly eliminated by the covalent binding of chondroitin sulfate to collagen. The cytotoxicity of residual glutaraldehyde--leaching through the interstices of the collagen fibrils or the tissue matrix--and of reactive aldehydes associated with the bound polymeric glutaraldehyde can be minimized by neutralization and thorough rinsing after crosslinking and storage in a nontoxic bacteriostatic solution. PMID- 3036882 TI - Toxic chemicals versus lung tissue--an aspect of inhalation injury revisited. The Everett Idris Evans memorial lecture--1986. PMID- 3036883 TI - Choledochocolonic fistula--a rare complication of cholangiocarcinoma. PMID- 3036884 TI - Tumor inhibition by titanocene complexes: influence upon two xenografted human lung carcinomas. AB - The influence of the organometallic complexes titanocene dichloride, titanocene dibromide, titanocene bis(hydrogenmaleinate), and titanocene bis(p aminothiophenolate) bis(hydrochloride) on the development of a lung adenocarcinoma and a small cell lung carcinoma, both xenografted into athymic mice, was investigated in the present study. The tumors were growing s.c., and the substances administered i.p. as fivefold injections according to a Q2D X 5 (every 2 days X 5) or a Q3D X 5 schedule. In the case of lung adenocarcinoma, titanocene complexes inhibited tumor growth by more than 50% resulting in treated/control values of 20%-50%. The suppression remained stable beyond the end of the treatment period. In the case of small cell lung carcinoma, only those Q2D X 5 schedules, which corresponded to LD10 doses, effected considerable and stable growth inhibition to less than 50% of control values. Titanocene dichloride showed the greatest activity against both human tumors, followed by the p aminothiophenolate, and hydrogenmaleinate derivatives. PMID- 3036885 TI - A clonal derivative of tunicamycin-resistant Chinese hamster ovary cells with increased N-acetylglucosamine-phosphate transferase activity has altered asparagine-linked glycosylation. AB - A population of Chinese hamster ovary (CHO) cells resistant to the antibiotic tunicamycin (TM) had previously been isolated (Criscuolo, B.A., and Krag, S.S. (1982) J. Cell Biol. 94:586-591) by a stepwise selection procedure using progressive increments of TM added to the medium. TM inhibits asparagine-linked glycoprotein biosynthesis by blocking the transfer of N-acetylglucosamine-1 phosphate from the sugar nucleotide UDP-N-acetylglucosamine to the isoprenoid lipid carrier, dolichyl phosphate. Four clonal derivatives were isolated from the TM-resistant population in the presence of 27 micrograms TM/ml and were found to overproduce the N-acetylglucosamine-phosphate transferase activity to the same extent (approximately 15-fold compared to wild-type cells). One of these clones, 3E11, was greater than 550-fold more resistant to TM than wild-type cells. The resistance phenotype remained during at least 2.5 months of growth in the absence of TM. 3E11 cells exhibited chromosomal translocations, but no homogeneously staining regions (HSR) or double minute chromosomes. The N-acetylglucosamine phosphate transferase activity in 3E11 cells was membrane-associated and was inhibited by TM. A 140,000-dalton membrane protein and at least four other membrane proteins were enriched in 3E11 cells. Mannosylphosphoryldolichol synthase and glucosylphosphoryldolichol synthase activities were not elevated in membranes prepared from 3E11 cells. Asparagine-linked glycosylation was altered such that 3E11 cells synthesized primarily a truncated oligosaccharide, Man5GlcNAc2, perhaps due to the reduced amount of mannosylphosphoryldolichol relative to wild-type cells. PMID- 3036886 TI - Effect of acute serum depletion on Na+-K+ homeostasis in cultured human skin fibroblasts. AB - In order to elucidate changes in cell transport behavior of cultured human skin fibroblasts in response to acute serum depletion, we performed uptake and washout of 22Na+ and 86Rb+ as well as measurements of the intracellular Na+ and K+ levels in the presence and absence of ouabain. Pronounced and lasting increase in cellular Na+ and decrease in K+ were observed after removal of fetal bovine serum (FBS) from the medium. The sum of the Na+ and K+ contents (nEq/10(5) cells) was lower in FBS-free medium (mean +/- SD; 17.3 +/- 2.2) than in FBS-containing medium (26.2 +/- 3.8; P less than .02). Simultaneously, a decrease in cellular water volume was detected in the FBS-free medium. The cation uptake and washout data suggest that FBS removal primarily renders the cells more permeable to Na+ and K+ with a secondary stimulation of the ouabain-sensitive Na+ extrusion mechanism. FBS at a concentration of 0.2% prevented approximately 50% of the maximal increase in the 86Rb+ washout rate constant associated with FBS depletion. Ouabain (2 microM) produced an increase in the 86Rb+ washout rate constant. This effect was substantially larger in cells subjected to medium without FBS (from 0.0303 to 0.2500 min-1) than in fibroblasts incubated in medium with FBS (from 0.0107 to 0.0487 min-1). The cellular K+ content was drastically reduced by ouabain to a level not different in medium with or without FBS (33.9 +/- 4.5 to 1.75 +/- 0.38 and 16.7 +/- 1.4 to 1.4 +/- 0.13 nEq/10(5) cells, respectively). The 22Na+ washout data exhibited a three-exponential pattern. Analytical solutions of the washout data by means of two models (serial and parallel) with three compartments showed that FBS depletion resulted in increase of the size of all three compartments. It is concluded that in cultured human skin fibroblasts, FBS is essential to the maintenance of a normal Na+ and K+ homeostasis. The removal of FBS results in dramatic permutation of this homeostasis that develops within minutes and lasts for hours. PMID- 3036887 TI - Epidermal growth factor receptor expression during morphological differentiation of pheochromocytoma cells, induced by nerve growth factor or dibutyryl cyclic AMP. AB - Rat pheochromocytoma cells (clone PC12) possess functional surface receptors for both nerve growth factor (NGF) and epidermal growth factor (EGF). PC12 cells respond to NGF as well as to dibutyryl cyclic AMP (dbcAMP) by arrest of cell proliferation and initiation of morphological differentiation, while EGF acts as a mitogen. Exposure of PC12 cells to NGF for several days resulted in a complete loss of rapid EGF responses, such as membrane ruffling and activation of active K+ transport. EGF binding studies revealed that this loss of EGF responses was due to an almost complete reduction of the number of EGF binding sites. In contrast, exposure of PC12 cells to dbcAMP for 2 days did not affect the rapid EGF responses, despite the morphological differentiation. Moreover, EGF binding studies demonstrated a twofold increase in the number of high-affinity binding sites and a small increase in the number of low-affinity sites. In addition, exposure of the cells to dbcAMP caused a twofold increase of EGF-receptor phosphotyrosine kinase activity. These results indicate that neither EGF-binding or the presence of EGF receptors nor the rapid EGF responses are sufficient for persistent proliferation, on one hand, or sufficient to avoid morphological differentiation, on the other. PMID- 3036888 TI - Cyclic AMP-mediated suppression of normal and neoplastic B cell proliferation is associated with regulation of myc and Ha-ras protooncogenes. AB - Cyclic AMP functions as a negative regulator of cell proliferation in a variety of cell systems. We show here that the proliferation of normal and neoplastic B cells can be inhibited by high intracellular levels of cAMP. Thus forskolin treatment of the neoplastic B precursor cell line Reh induced a rapid increase in the cAMP level, which was followed by an accumulation of cells in the G0/G1 phase of the cell cycle over a period of 2-3 days. Similar inhibition of Reh cell proliferation after 3 days was observed whether forskolin was present continuously or only during the first 5 hr. Both c-myc and c-Ha-ras protein levels were transiently down-regulated at 4 hr of forskolin treatment, suggesting that these protooncogenes play a role in the process leading to cAMP-mediated growth cessation. Northern-blot analysis showed that the steady-state levels of c myc RNA rapidly declined in all phases of the cell cycle, to return to control levels within a time period of 24 hr. In contrast, the c-Ha-ras mRNA level was steadily maintained. Thus the expression of c-myc and c-Ha-ras protein was regulated at different metabolic levels. The reduced proliferative capacity of the B precursor cell line in the presence of forskolin was not linked to induced differentiation. This was judged from the lack of appearance of three different B cell differentiation markers; cytoplasmic immunoglobulin heavy chain and two antigens recognized by the monoclonal antibodies B1 (CD20) and HH1 (CD37). We also showed that forskolin partially inhibited the proliferation of normal B lymphocytes stimulated by anti-immunoglobulins (anti-mu) and B cell growth factor (BCGF). The burst of c-myc mRNA during activation of normal B cells was also reduced by forskolin. PMID- 3036889 TI - Effect of phagocytosis on receptor distribution and endocytic activity in macrophages. AB - Phagocytosis requires the internalization of a significant fraction of the plasma membrane and results in the intracellular deposition of large particles. We evaluated the effect of phagocytosis on the cellular distribution of recycling receptors and uptake of ligand to determine whether phagocytosis affects receptor behavior. Phagocytosis of zymosan, latex particles, or IgG-coated red blood cells by rabbit alveolar macrophages did not decrease the number of cell surface receptors for transferrin, alpha 2-macroglobulin X protease complexes, maleylated proteins, or mannosylated proteins. The number of surface receptors for transferrin was also unaltered in J774 cells, a macrophage-like cell line. In both cell types extensive phagocytosis did not affect the rate of receptor mediated endocytosis or the distribution of receptors between the endosome and the cell surface. However, fluid phase pinocytosis was reduced by phagocytosis. The major reduction appeared to be not in the rate of internalization but rather in the delivery of fluid to the lysosome. These results demonstrate that internalization of a significant amount of the plasma membrane during phagocytosis does not diminish the number of receptors on the cell surface and has no effect on receptor-mediated ligand uptake. PMID- 3036890 TI - Down-modulation of EGF receptors in cells transformed by the src oncogene. AB - The effects of src oncogene expression on epidermal growth factor (EGF) receptors have been investigated in mouse 3T3 and rat-1 fibroblasts. Transformation of both cell types with src resulted in marked reductions in cellular EGF receptor levels, as assayed by either 125I-EGF binding or immunoprecipitation of receptor protein from radiolabeled cell lysates. In contrast to cells transformed by other types of retroviral oncogenes, the loss of EGF receptors in the src-transformed cells did not appear to be due to secreted transforming growth factor-alpha (TGF alpha), since such factors were undetectable in culture fluids from the src transformed cells. By several criteria of transformation, an EGF-receptorless cell line infected with src was shown to be transformed, suggesting that EGF receptors themselves are not obligatory to the src transformation process. We suggest that pp60src down-modulates EGF receptors by an intracellular mechanism and that the loss of the receptors is symptomatic of more general effects of pp60src on the machinery of growth regulation. PMID- 3036891 TI - Stimulation by serum of the Na+/H+ antiporter in quiescent pig kidney epithelial (LLC-PK1) cells and role of the antiporter in the reinitiation of DNA synthesis. AB - LLC-PK1 cells can be brought into a classical quiescent state by depriving them of serum for 6 days. At this time, pulse-labeling with [3H]-thymidine shows that only 3% of the cells are synthesizing DNA, but the quiescent cells can be stimulated with serum to re-enter the cell cycle at a point early in G1. The rate of amiloride-sensitive 22Na+ uptake (as a measure of the Na+/H+ antiporter) is relatively low during quiescence; it rises 2- to 3-fold within 4 h after serum addition. This increase in antiporter activity appears to be required for the resumption of DNA synthesis in the absence of bicarbonate, because ethylisopropylamiloride (EIPA) blocks [3H]-thymidine incorporation when serum is added to cells in bicarbonate-free medium. In the presence of bicarbonate, however, EIPA has no effect on [3H]-thymidine incorporation, indicating that another (bicarbonate-dependent) transport system can substitute for the antiporter under these conditions. PMID- 3036892 TI - Macrophage cell lines transformed by the malignant histiocytosis sarcoma virus: increase of CSF receptors suggests a model for transformation. AB - The malignant histiocytosis sarcoma virus (MHSV) contains Ha-v-ras-related oncogenic sequences and rapidly transforms myeloid cells in vivo and in vitro. Myeloid cell lines can be derived which do not require growth factor for continued proliferation. We initiated this work to define the process of transformation leading to autonomy of cell growth in transformed myeloid cells. Five established cell lines were examined. All express macrophage-specific cell surface antigens and exhibit several other properties typical for mature macrophages. Growth properties, growth factor release, and growth factor receptor presentation were examined: Release of growth factors is not a consistent feature. All cell lines show cell-density-independent colony formation and do not release self-stimulating factors, thus excluding autocrine stimulation as a model leading to transformation. All cell lines express unusually high levels of granulocyte-macrophage (GM)- and multi-CSF receptors and, except for one, M-CSF receptors. The high increase in GM-CSF and other growth factor receptors may be causally related to the transformed state of the cells. MHSV can be used as a tool to easily derive cell lines of the macrophage pathway as a model to study myeloid transformation, differentiation, and macrophage function. PMID- 3036893 TI - Possible involvement of reorganization of actin filaments, induced by tumor promoting phorbol esters, in changes in colony shape and enhancement of proliferation of cultured epithelial cells. AB - Tumor promoters are known to induce reorganization of actin, morphological changes and enhancement of proliferation of epidermal cells in vivo. In this study, we have examined the effects of tumor promoters on these events to clarify the role played by the organization of actin filaments in the regulation of the shape and growth of colonies of epithelial cells in culture. Treatment with 12-O tetradecanoylphorbol-13-acetate (TPA) caused a change in the shape of colonies of FL and Madin-Darby canine kidney (MDCK) cells within 6 hr. Changes in the shape of colonies were consistent with the morphological change of individual cells and the dissociation of groups of cells in the colonies. Addition of TPA also caused reorganization of actin filaments after 2 hr, and it caused enhancement of proliferation of FL and MDCK cells after 48 hr but did not cause any such changes in KB cells. However, the binding affinities of 4 beta-phorbol 12,13-dibutyrate (PDBu) to FL and MDCK cells were similar to that of PDBu to KB cells. Related tumor promoters such as phorbol 12,13 didecanoate (PDD) and mezerein caused effects similar to those caused by TPA. In contrast, nontumor promoting phorbol esters, such as 4 alpha-PDD and phorbol, had no effect. Cyclic AMP blocked the TPA-induced changes in FL and MDCK cells. These results suggest that TPA-induced reorganization of actin filaments which can be inhibited by cyclic AMP results in changes in the shape of colonies and enhancement of proliferation. PMID- 3036894 TI - Na+ for H+ exchange in rabbit erythrocytes. AB - The effect of a transmembrane pH gradient on the ouabain, bumetanide, and phloretin resistant H+ efflux was studied in rabbit erythrocytes. Proton equilibration was reduced by the use of DIDS (125 microM) and acetazolamide (1 mM). H+ efflux from acid loaded erythrocytes (pHi = 6.1) was measured in a K+ (145 mM) medium, pH0 = 8.0, in the presence and absence of 60 microM 5,N,N dimethyl-amiloride (DMA). The H+ efflux rate in a K+-containing medium was 116.38 +/- 4.5 mmol/l cell X hr. Substitution of Nao+ for Ko+ strongly stimulated H+ efflux to 177.89 +/- 7.9 mmol/l cell X hr. The transtimulation of H+ efflux by Nao+ was completely abolished by DMA falling to values not different from controls with an ID50 of about 8.6 X 10(-7) M. The sequence of substrate selectivities for the external transport site were Na greater than greater than greater than Li greater than choline, Cs, K, and Glucamine. The transport system has no specific anion requirement, but is inhibited by NO3-. The DMA sensitive H+ efflux was a saturable function of [Na+]o, with an apparent Km and Vmax of about 14.75 +/- 1.99 mM and 85.37 +/- 7.68 mmol/l cell X hr, respectively. However, the Nao+-dependent and DMA-sensitive H+ efflux was sigmoidally activated by [H+]i, suggesting that Hi+ interacts at both transport and modifier sites. An outwardly directed H+ gradient (pHi 6.1, pH = 8.0) also promoted DMA sensitive Na+ entry (61.2 +/- 3.0 mmol/l cell X hr) which was abolished when pHo was reduced to 6.0. The data is therefore consistent with the presence of a Na+/H+ exchange system in rabbit erythrocytes. PMID- 3036895 TI - Types I and IV collagenolytic and plasminogen activator activities in preovulatory ovarian follicles. AB - During ovulation, enzymatic degradation of the extracellular matrix occurs within and around the graafian follicles. In this study, the activities of several different proteolytic enzymes were measured in the culture media of follicles taken from pregnant mare serum gonadotropin (PMSG)-primed immature rats. At 52 h after PMSG, the follicles were cultured for 2 to 15 h in media with or without human chorionic gonadotropin (hCG). Type I collagenase activity in hCG-stimulated follicles gradually increased within 6 h to 3.3-fold above that of the controls. Relatively pure populations of granulosa cells produced type I collagenase to a similar extent. Likewise, type IV collagenase increased 3.8-fold by 6 h after exposure of the follicles to hCG. In contrast, plasminogen activator activity increased by 3.9-fold at 2 h after hCG, but was negligible at 4, 6, and 15 h after incubation. These results suggest that plasminogen activator may activate both type I and type IV collagenase in hCG-stimulated ovulatory follicles. PMID- 3036897 TI - Effects of pretreatment on the enantioselectivity of silica-bound bovine serum albumin used as high-performance liquid chromatographic stationary phases. PMID- 3036896 TI - Regulation of the ubiquitin-mediated proteolytic pathway: role of the substrate alpha-NH2 group and of transfer RNA. AB - Degradation of intracellular proteins via the ubiquitin pathway involves several steps. In the initial event, ubiquitin becomes covalently linked to the protein substrate in an ATP-requiring reaction. Following ubiquitin conjugation, the protein moiety of the adduct is selectively degraded with the release of free and reusable ubiquitin. Ubiquitin modification of a variety of protein targets in the cell plays a role in basic cellular functions. Modification of core nucleosomal histones is probably involved in regulation of gene expression at the level of chromatin structure. Ubiquitin attachment to cell surface proteins may play roles in processes of cell-cell interaction and adhesion, and conjugation of ubiquitin to other yet to be identified protein(s) could be involved in the progression of cells through the cell cycle. Despite the considerable progress that has been made in the elucidation of the mode of action and cellular roles of the ubiquitin pathway, many major problems remain unsolved. A problem of central importance is the specificity in the ubiquitin ligation system. Why are certain proteins conjugated and committed for degradation, whereas other proteins are not? A free alpha-NH2 group is an important feature of the protein structure recognized by the ubiquitin conjugation system, and tRNA is required for the conjugation of ubiquitin to selective proteolytic substrates and for their subsequent degradation. These findings can shed light on some of the features of a substrate that render it susceptible to ubiquitin-mediated degradation. PMID- 3036899 TI - Thermospray collisionally induced dissociation with single and multiple mass analyzers. AB - A thermospray discharge ionization source has been used to obtain a collisionally induced dissociation (CID) spectrum of nabilone (MW 372) using a single stage quadrupole mass spectrometer. The resulting CID-mass spectrometry (MS) data show only a few structural features. Since most of the important fragments are above mass 200, it is difficult to assign structural identities to the primary daughter fragments or the nabilone molecule. Further information was obtained by using the thermospray discharge ionization source with a triple stage quadrupole mass spectrometer. Selected primary daughter ions were subjected to a second stage of CID fragmentation. Many of the resulting granddaughter ions are now in the mass range of 50-150 daltons and confident structure assignments can be made. The CID MS-CID-MS data on nabilone showed the following features: a C9 aliphatic chain with a branch giving a C6 chain; losses of masses 58, 86, and 100 suggesting a ketonic structure that can break with up to 6 aliphatic carbons; losses of masses 60, 61, and 102 suggesting an ether or alcoholic oxygen; presence of one aromatic ring with one or two phenolic oxygens; low mass fragments of masses 55, 69, and 83 indicative of rings or unsaturation. It is audacious to suggest that the information given here is sufficient to write the full structure of nabilone. Nevertheless, the granddaughter fragment information permits a reasonable reconstruction of possible structures that could not be made with the first-order collisional fragmentation. PMID- 3036898 TI - Enrichment of complexing analytes on a copper-loaded silica surface and on-line coupling with high-performance liquid chromatography, using L-tryptophan and L tyrosine as model compounds. AB - A chemically modified silica (bisdithiocarbamate) strongly binds copper and has a loading capacity of 0.5 mmol g-1 for the copper ion. This new metal-loaded silica, packed in a short stainless-steel pre-column, is highly selective for the enrichment of two amino acids chosen as model compounds. On-line coupling of this pre-column with an analytical RP-18 column allowed the separation and detection of L-tryptophan and L-tyrosine at parts per billion levels in aqueous media with good accuracy. Separation was achieved by ion-pair chromatography and the solutes were detected by fluorescence. The ions generally present in natural aqueous media did not interfere. PMID- 3036900 TI - Direct analysis of the major human seminal prostaglandins by thermospray high performance liquid chromatography-mass spectrometry. AB - Thermospray high-performance liquid chromatography-mass spectrometry (HPLC-MS) can be a powerful tool for characterizing eicosanoids in complex biological samples. The positive ion spectra obtained from primary prostaglandins such as PGE1 PGE2, 19-OHPGE1, 19-OHPGE2, PGF2 alpha, PGD2, 6-keto-PGF1 alpha and from leukotriene B4 are very simple, with base peaks corresponding to ions arising from the loss of H2O from the (M + H)+ and (M + NH4)+ ions, except for PGB2 and PGF2, where the latter two ions predominate. The application of this technique to the concurrent determination of the E1 and E2 prostaglandins and their 19 hydroxylated derivatives in human semen is described. The technique affords a moderate level of sensitivity (5-20 ng on-column) and excellent specificity so that virtually no sample manipulation is required other than dilution in acetone and centrifugation. The clear supernatant is injected directly into the HPLC-MS system. A similar analysis by either gas chromatography (GC) or GC-MS would need multi-step derivatization, thus increasing the sample manipulation required and the total analysis time. PMID- 3036901 TI - Determination of leukotriene B4 by high-performance liquid chromatography with electrochemical detection. PMID- 3036902 TI - Adrenal androgen response to metyrapone, adrenocorticotropin, and corticotropin releasing hormone stimulation in children with hypopituitarism. AB - We determined the adrenal steroid responses to metyrapone, ACTH, and CRH in 12 ACTH-intact and 5 ACTH-deficient hypopituitary children to determine the mechanisms that control adrenal androgen secretion. Serum adrenal androgen concentrations [dehydroepiandrosterone (DHEA) and delta 4-androstenedione (delta 4-A)] rose in response to oral administration of metyrapone (450 mg/m2 X dose, every h for 7 doses) in ACTH-intact hypopituitary children with multiple or isolated pituitary hormone deficiencies [mean postmaryrapone level: DHEA, 225 ng/dL (range, 27-566); delta 4-A, 313 ng/dL (range, 105-651)], except in 2 young children in whom DHEA did not rise. These adrenal androgens did not rise in all ACTH-deficient hypopituitary children [mean postmetyrapone level: DHEA, 11.0 ng/dL (range, 3-16); delta 4-A, 6.2 ng/dL (range, 3-10)]. The increases in both serum cortisol and adrenal androgens, including DHEA sulfate, in response to short term ACTH infusion (40 U in 6 h) in ACTH-intact hypopituitary children were normal or above normal, while these steroid responses were significantly (P less than 0.05-0.01) lower in ACTH-deficient hypopituitary children compared to normal values. However, prolonged administration of ACTH (40 U/day, or im) for 6 days to 2 ACTH-deficient hypopituitary children resulted in normal DHEA responses to the 6-h ACTH stimulation test (DHEA levels after the first test, 14 and 30 ng/dL, after priming, 80 and 50 ng/dL). Furthermore, CRH administration to 4 ACTH deficient patients caused a rise in serum DHEA and cortisol in patients with a normal ACTH response, while those with a poor ACTH response had a lesser rise in DHEA and cortisol. These data suggest that ACTH is the major tropic hormone for adrenal androgen secretion. PMID- 3036903 TI - Inhibition of the adrenocorticotropin response to surgery in humans: interaction between dexamethasone and fentanyl. AB - We examined the plasma ACTH and cortisol responses to surgery in 25 patients with atherosclerotic heart disease undergoing myocardial revascularization. The patients were all premedicated with diazepam, and general anesthesia was induced with thiopental. They were randomly assigned to one of four groups: I) no dexamethasone (DEX), enflurane anesthesia, II) 40 mg DEX, iv, 45-60 min before sternotomy, enflurane anesthesia, III) no DEX, fentanyl [N-(1-phenethyl-4 piperidyl)propionanilide] anesthesia (50-100 micrograms/kg), and IV) DEX, fentanyl anesthesia. Isokalemic hemodilution of significant magnitude occurred during cardiopulmonary bypass. All groups had significant increases in plasma ACTH during surgery, which returned to control levels 22 h after the bypass. Group I (no DEX, no fentanyl) and group III (no DEX, fentanyl) patients had large similar increases in plasma ACTH, which peaked 2-4 h postbypass [400 +/- 83 (+/- SEM) pg/mL; 88 +/- 18 pmol/L]. The group II (DEX, no fentanyl) patients also had large increases in ACTH which were similar to those in groups I and III, except 2 4 h postbypass (183 +/- 91 pg/mL; 40 +/- 20 pmol/L). The group IV (DEX, fentanyl) patients had a significantly attenuated ACTH response to surgery; the mean plasma ACTH level 2-4 h postbypass was only 54 +/- 21 pg/mL (12 +/- 5 pmol/L). Therefore, although DEX or fentanyl alone had a minimal effect on the ACTH response to surgery, a significant attenuation occurred when DEX and fentanyl were used in combination. We conclude that glucocorticoids and morphine agonists exert interactive inhibitory effects on ACTH release in humans, probably by virtue of their suppression of CRH release from the hypothalamus. PMID- 3036904 TI - Direct stimulation of nucleoside triphosphatase activity in human ovarian nuclear membranes by human chorionic gonadotropin. AB - We previously reported that nuclei isolated from ovaries of premenopausal women contain binding sites for hCG/human LH (hLH). This study was undertaken to determine the possible functional significance of these nuclear binding sites. Upon addition to isolated ovarian (mostly luteal cells) nuclear membranes, hCG and hLH stimulated nucleoside triphosphatase (NTPase), an enzyme involved in nucleocytoplasmic transfer of mRNA, but not Mg2+-ATPase or NADH cytochrome c reductase activities, in a concentration-dependent manner. Heat-denatured hCG, isolated alpha- and beta-subunits of hCG, human FSH, PRL, and porcine relaxin had no effect on the enzyme. Addition of hCG antiserum blocked hCG's ability to stimulate NTPase activity. cAMP, which is a second messenger in hCG- and hLH stimulated steroidogenesis, had no effect on NTPase activity. These results, which demonstrate that hCG acts on human ovarian nuclei directly, raise the possibility that internalized hCG might influence nuclear function(s) before it is eventually degraded in the lysosomes of ovarian cells. PMID- 3036905 TI - Urinary 18-hydroxycortisol and its relationship to the excretion of other adrenal steroids. AB - The urinary excretion of 18-hydroxycortisol was recently reported to be increased in patients with primary aldosteronism who have an adrenal adenoma and in those with glucocorticoid-suppressible aldosteronism. A direct RIA for 18 hydroxycortisol in urine and plasma has been described, and we here report our experience using a similar direct RIA and a more elaborate RIA which includes a preliminary high pressure liquid chromatography (HPLC) purification step. The urinary excretion of 18-hydroxycortisol was compared with that of other adrencorticoids. The urinary excretion of 18-hydroxycortisol in 37 normal subjects using the direct RIA was 112 +/- 49 (+/- SD) microgram/24 h, and that with the HPLC-RIA method was 63 +/- 36 micrograms/24 h. The accuracy and specificity of the HPLC-RIA assay method were confirmed by measuring the steroid after the HPLC step as the glycolic acid ester derivative. The urinary excretion of 18-hydroxycortisol correlated with that of cortisol (r = 0.36; P less than 0.01), 18-oxocortisol (r = 0.42; P less than 0.01), and 19-nordeoxycortisosterone (r = 0.71; P less than 0.001), but did not correlate with the excretion of aldosterone 18-oxoglucuronide (r = 0.25; P = 0.15942). Dexamethasone administration to five normal subjects significantly decreased 18-hydroxycortisol excretion from 81 +/- 47 to 23 +/- 8 micrograms/24 h. ACTH infusion in these subjects receiving dexamethasone significantly raised 18-hydroxycortisol excretion to 147 +/- 37 micrograms/24 h. Five days of a sodium-restricted diet (10 mmoles/day) resulted in a significant (P less than 0.02) increase in 18 hydroxycortisol excretion, but two of eight subjects had decreased excretion, although urinary aldosterone excretion increased, as expected. These studies demonstrate that the direct RIA significantly overestimates urinary 18 hydroxycortisol excretion. These studies also demonstrate that the major factor resulting 18-hydroxycortisol excretion is ACTH. However, since 18-hydroxycortisol excretion may increase during sodium depletion, angiotensin or other factors may also regulate its secretion. PMID- 3036907 TI - Cyclic adenosine-3',5'-monophosphate alters hydrolysis of phospholipids by mouse embryo palate mesenchyme cells. AB - The purpose of this study was to determine whether inhibition of release of arachidonic acid from mouse embryo palate mesenchyme (MEPM) cells in response to cAMP is due to a selected or generalized inhibition of hydrolysis of esterified pools of arachidonic acid. The calcium ionophore A23187 proved to be a useful probe of phospholipid hydrolases in MEPM cells, since it stimulated release of radiolabeled fatty acids from phospholipids of prelabeled MEPM cells as a function of the length of exposure, concentration, and concentration of Ca2+ in the medium. Elevation of intracellular levels of cAMP by treatment with (-) isoproterenol resulted in the inhibition of release of radiolabeled arachidonic acid in response to A23187. Analysis by quantitative gas-liquid chromatography revealed the source of the arachidonic acid released in response to the ionophore to be 1,2-diradyl-sn-glycero-3-phosphoethanolamine; elevation of intracellular levels of cAMP inhibited hydrolysis of this substrate, but may have stimulated hydrolysis of 1,2-diradyl-sn-glycero-3-phosphocholine. These findings permit the conclusions that 1) the ionophore stimulates activities of selected phospholipases A in MEPM cells and 2) cAMP modulates certain phospholipases A in MEPM cells in a specific manner. PMID- 3036906 TI - Immunopathogenesis and treatment of myasthenia gravis. PMID- 3036908 TI - Protection between different serotypes of bovine rotavirus in gnotobiotic calves: specificity of serum antibody and coproantibody responses. AB - In a previous study, different U.S. isolates of bovine rotavirus were studied for their serotypes and cross-protective properties (G. N. Woode, N. E. Kelso, T. F. Simpson, S. K. Gaul, L. E. Evans, and L. Babiuk, J. Clin. Microbiol. 18:358-364, 1983). Three viruses belonging to two different serotype groups were used as vaccines in gnotobiotic calves, which were subsequently challenged with B641 or B223, representing the two bovine serotypes. In the present work, the experiments were repeated with more calves and the specificity of their antibody responses was measured and compared with the results of the protection studies. Protection between different serotypes occurred under both homologous and heterologous conditions but was not directly serotype dependent. B223 virus showed both homologous and heterologous protection against B223 and B641 challenge viruses. This was a one-way reaction, as B641 did not induce protection against B223. Neonatal calf diarrhea virus vaccine produced neither homologous (against B641) nor heterologous (against B223) protection. The plaque reduction neutralization titers of serum antibody and coproantibody did not predict a state of protection against the challenge virus. Calves vaccinated with neonatal calf diarrhea virus or B641 developed neutralizing antibodies to their respective heterologous challenge viruses but were not protected. After challenge, the boosted coproantibody plaque reduction neutralization response to the original vaccine virus was greater than that to the challenge virus. PMID- 3036909 TI - Comparative evaluation of a commercial enzyme-linked immunoassay and solid-phase immune electron microscopy for rotavirus detection in stool specimens. AB - Using solid-phase immune electron microscopy (SPIEM) as a reference test, we examined 151 stool specimens from infants and young children with acute gastroenteritis for rotavirus detection by a one-step commercial enzyme-linked immunosorbent assay (ELISA) with labeled monoclonal antibody. Of the 83 samples determined to be positive for rotavirus by SPIEM, 82 were detected as positive by the monoclonal antibody ELISA (sensitivity, 98.7%), while 67 of the 68 specimens determined to be negative by SPIEM were correctly detected as negative by the ELISA (specificity, 98.5%). The diagnostic accuracy of the ELISA kit was 98.6%. Thus, the one-step monoclonal antibody ELISA, which can be completed in less than 90 min, appears to be highly suitable for the rapid and reliable detection of rotavirus in stools. PMID- 3036910 TI - Bovine milk immunoglobulins for passive immunity to infantile rotavirus gastroenteritis. AB - Pregnant cows were successfully hyperimmunized with all four human rotavirus serotypes, resulting in a 100-fold increase in neutralizing milk antibody titers over those of controls. Milk antibodies were isolated batchwise from 1,000 kg of pooled milk for the first 10 lactation days, yielding 10 kg of freeze-dried milk immunoglobulin concentrate consisting of 50% bovine milk immunoglobulins. Milk immunoglobulin concentrate showed neutralizing activities against all four human rotavirus serotypes that were 100 times higher than those in pooled human milk samples and 10 times higher than those in a commercial pooled immunoglobulin preparation from pooled human blood serum. In vitro neutralization tests showed that milk immunoglobulin concentrate had powerful antiviral activity, even against very high doses of infectious rotaviruses. Because the technology of the milk immunoglobulin concentrate ensures that it is innocuous and can be used for oral application, it is proposed that milk immunoglobulin concentrate be used to induce passive immunity to infantile rotavirus gastroenteritis. PMID- 3036912 TI - Axial heterogeneity of intracellular pH in rat proximal convoluted tubule. AB - In the proximal convoluted tubule (PT), the HCO3- reabsorptive rate is higher in early (EPS) compared with late proximal segments (LPS). To examine the mechanism of this HCO3- reabsorption profile, intracellular pH (pHi) was measured along the superficial PT of the rat under free-flow and stationary microperfusion using the pH-sensitive fluorescence of 4-methylumbelliferone (4MU). With 4MU superfusion, pHi was found to decline along the PT. Observation with 365-nm excitation revealed that EPS were brightly fluorescent and always emerged away from their star vessel. Midproximal segments were darker and closer to the star vessel which was surrounded by the darkest LPS. Decreasing luminal HCO3- from 15 to 0 mM lowered pHi in both EPS and LPS, but pHi remained more alkaline in EPS with both perfusates. Thus the axial decline in pHi along the PT is due to both luminal factors and intrinsic differences in luminal H+ extrusion in PT cells. PMID- 3036914 TI - Characterization of four herpesviruses isolated from owl monkeys and their comparison with Herpesvirus saimiri type 1 (Herpesvirus tamarinus) and herpes simplex virus type 1. AB - Four herpesviruses were previously isolated from four outbreaks of lethal disease in owl monkeys. All four isolates have been shown to be antigenically closely related to each other and to Herpesvirus saimiri 1 (HVS-1) by kinetic neutralizations. The owl monkey strains also share similarities to HVS-1 and to each other with respect to host range, growth cycles and molecular weights of peptides and of DNA fragments generated by restriction endonuclease (R.E.) digestion. R.E. analysis, however, can differentiate strains by the use of certain enzymes. All four isolates share a common G-C ratio percentage with HVS-1 of 67 per cent. On the basis of these findings, we believe that these owl monkey virus isolates are strains of HVS-1. PMID- 3036913 TI - Pattern of lateral geniculate synapses on neuron somata in layer IV of the cat striate cortex. AB - The distribution of geniculate synapses on neuron cell bodies in layers IVab and IVc of cat area 17 was studied. Electron microscope autoradiography was used to identify geniculate terminals that were labeled by anterograde transport of radioactivity injected into the A-laminae of the lateral geniculate nucleus. Thirty-eight cell bodies (19 in layer IVab and 19 in layer IVc) were examined in a series of 138 consecutive sections. Two pyramidal somas were studied and had no geniculate contacts. All of the other somas studied were nonpyramidal, and of these, 85% received geniculate contacts. The proportion of somas receiving somatic geniculate input differed in layers IVab and IVc. In layer IVab, 70% of the nonpyramidal somas received geniculate contacts; in IVc, 100%. Such high percentages indicate that geniculate afferents synapse with more types of layer IV neuron than the aspinous neurons that synthesize gamma-aminobutyric acid (GABA) (Freund et al., '85b). The pattern of input to somas was so diverse that it was impossible to form groups of neurons based on only this criterion. We wondered if it would be possible to form groups of neurons based on a range of characteristics among which would be pattern of synaptic input. To this end, pyramidal neurons and neurons that contained a cytoplasmic laminated body (CLB) (Winfield, '79; Einstein et al., '84) were treated as two separate classes. We found fair agreement among the features of these neurons within their own classes, with the CLB-cells in layer IVab and IVc forming separate groups. Among the remaining neurons there was too little agreement within the range of features to enable us to treat them in this manner. Geniculate somatic contacts in both sublayers were of 2 forms, those with round vesicles and asymmetric thickenings (RA) and those with pleomorphic vesicles and symmetric thickenings (PS) (Einstein et al., '87). The distribution of these forms varied: some cells received contacts exclusively from one form or the other; other cells received contacts from both. On one cell that bore 33 somatic geniculate terminals, 61% were RA and 39% were PS. Such substantial numbers of geniculate contacts located near the site of impulse initiation are likely to contribute significantly to the receptive field properties of this neuron, and the possible effects are discussed. PMID- 3036911 TI - Relationship of phosphatidylinositol bisphosphate hydrolysis to calcium mobilization and functional activation in fluoride-treated neutrophils. AB - Sodium fluoride (20 mM) effected rapid hydrolysis of phosphatidylinositol bisphosphate (PIP2) in human neutrophils. Intracellular free Ca2+ levels increased after PIP2 hydrolysis but before respiratory burst activation. Both the increase in intracellular free Ca2+ levels and the extent of functional activation were dependent on the availability of extracellular Ca2+. The rate of F(-)-stimulated PIP2 hydrolysis, however, was not affected when the rise in cytosolic Ca2+ was severely limited by depletion of extracellular Ca2+. Fluoride caused the specific hydrolysis of PIP2 in isolated neutrophil plasma membranes. This effect occurred in the presence of low levels of available Ca2+ and was accompanied by the release of inositol phosphates. We conclude that PIP2 hydrolysis is an early event in the response of neutrophils to F-. This response is not Ca2+-regulated but may lead to an influx of Ca2+ from the extracellular medium. Activation of a PIP2-specific phospholipase independent of a change in cytosolic free Ca2+ levels may be the initial event in the stimulus-response pathway triggered by fluoride. PMID- 3036915 TI - Complex viral and fungal skin lesions of patients with acquired immunodeficiency syndrome. AB - Three patients with acquired immunodeficiency syndrome were noted to have skin lesions in which there were histologic changes supporting the presence of both fungal and viral pathogens. Skin lesions were found in the patellar region in two patients and in the groin in one patient. Skin biopsies revealed massive epidermal hyperplasia, cytopathic changes of herpesvirus infection in all three patients, papillomavirus in one patient, and septate hyphae in the stratum corneum in all patients. The coexistence of fungal and viral infection produces an epidermal response that in some features resembles that seen in keratoacanthoma. PMID- 3036916 TI - MR imaging of the thyroid: comparison with scintigraphy in the normal and diseased gland. AB - Spin-echo magnetic resonance (MR) imaging of the thyroid gland was performed in patients using a superconducting magnet operating at 0.35 T. There were 17 women and two men with an age range of 21-77 years. All of the patients with disease were also evaluated with scintigraphy and three of the four subjects with normal thyroid glands also had scintigraphy. Final diagnoses in the patients were normal gland in four, Graves disease in two, thyroid cyst in one, benign follicular adenoma in two, papillary cell carcinoma in one, Hurthle cell carcinoma in one, Hashimoto thyroiditis in one, and multinodular goiter in seven. The normal thyroid and surrounding anatomy were clearly demonstrated by intrinsic signal intensity differences of the tissues. The thyroid gland in Graves disease was enlarged and had homogeneously increased signal intensity at all pulse sequences compared with skeletal muscle and normal thyroid. The thyroid cyst and benign adenoma were well defined; however, the cyst displayed the greater signal intensity on more T2-weighted pulse sequences. The MR signal intensity features of multinodular goiter and Hashimoto thyroiditis appear similar to those of the normal gland. In cases of focal masses, MR could not reliably distinguish benign from malignant tumor. However, using intensity ratio data there was a statistically significant difference between solid and hemorrhagic cystic disease. PMID- 3036917 TI - Benign pleomorphic adenomas of the salivary glands: surface coil MR imaging versus CT. AB - High resolution magnetic resonance (MR) imaging of three salivary gland tumors was performed at 1.5 T and compared with CT. Two of the three tumors were well seen on CT. However, one required CT sialography to separate tumor from the remaining parotid parenchyma, and, in the other, soft tissue invasion could not be excluded on the basis of CT. Magnetic resonance imaging demonstrated sharp tumor margins with no evidence of invasion. All three tumors were pathologically proven benign pleomorphic adenomas confirming the MR findings. On T2 weighted images, the two large adenomas demonstrated inhomogeneity that was not observed on CT. PMID- 3036918 TI - Tubular apocrine adenoma. AB - We report a case of tubular apocrine adenoma located on the scalp, with characteristics of syringocystadenoma papilliferum in the superior part of the lesion. An interesting feature of the growth is its connective tissue involvement. PMID- 3036919 TI - ACTH initiation of mammary secretion in pregnant goats is influenced by the stage of gestation and pre partum milking. AB - Pregnant goats were treated with ACTH by intramuscular injection (1 mg/d for 2 d) at various stages of pregnancy to investigate its effect on initiation of mammary secretion. ACTH stimulated lactose secretion and increased udder volume at or after 84 d of gestation, when compared with untreated controls. Treatment with ACTH between d 109 and 127 of gestation increased circulating plasma glucose concentration, mammary blood flow and mammary uptake of glucose which returned to pre injection levels 6 d after treatment. Arterial concentrations of progesterone were not affected by ACTH but mammary progesterone uptake increased 2-fold. Pre partum milking alone stimulated mammary secretion in eight of 13 goats. Subsequent treatment with ACTH resulted in no further change in the composition of the secretion but increase in yield was maintained. ACTH stimulated mammary secretion in those goats which did not respond to pre partum milking. It was concluded that ACTH initiated mammary secretion in pregnant goats by mechanisms which interact with the stimulation of milking but which are independent of circulating progesterone levels. PMID- 3036920 TI - Supplying the energy and fiber needs of dairy cows from alternate feed sources. AB - Alternate feeds are a major resource of the dairy industry. The major issue involving them is a method to predict accurately nutritive value from laboratory analyses. Variation in nutrient content of most alternate feeds is greater than in feed grains. Another issue is which depression factors to use in adjusting values for TDN from maintenance to production intakes. The NRC uses an average depression of 8% for all feeds; others think each feedstuff should be depressed individually, and discount factors have been proposed. For some alternate feeds, large differences in net energy estimates occur. Neutral detergent fiber has been proposed as an indicator of productive energy, but it has several deficiencies with alternate feeds high in fat, molasses, or ash. A summative equation based on fat, ash, protein, NDF, and lignin has wider application for predicting NE1 for all feeds. A roughage value index reflects a feed's property to stimulate chewing and rumination. Its use has special relevance for alternate feeds with small particle sizes, which may induce little chewing. Supplemental fat may increase the metabolizable energy converted to milk, but respiration experiments are needed. PMID- 3036922 TI - Outcomes of discharge from hospital for elderly people. AB - The numbers of elderly people going home from hospital in the United Kingdom are likely to increase over the next decade. Previous research suggests that this group of people are liable to experience some difficulties on returning home. This paper reports the findings of a study of the outcomes of discharge for 32 elderly people admitted to geriatric wards. A lower level of independence in daily living activities, and an increase in the amount of help needed after hospitalization were found. An increase in the rate of discharge of elderly people will have implications for both formal and informal carers. PMID- 3036921 TI - [Formation of peroxide-type radicals during low-temperature cell and tissue irradiation]. PMID- 3036923 TI - Monoamine transport in depression: kinetics and dynamics. AB - There appears to be an abnormality in monoamine transport in major depressive disorders that may be specific to these disorders, although it may not be primarily an aminergic lesion. Kinetic factors alone are insufficient to explain the observed effects of antidepressants on transport mechanisms, or changes in uptake with therapeutic response, suggesting dynamic influences on uptake processes that deserve further exploration. The investigation of these influences may indicate that platelet 5-HT uptake offers diagnostic and theoretical possibilities not adverted to at present and another rationale for the use of platelets in research on mental disorders. PMID- 3036924 TI - Maintenance of alveolar bone through implantation of bone graft substitutes in tooth extraction sockets. PMID- 3036925 TI - Relation of coronary artery stenosis and pressure gradient to exercise-induced ischemia before and after coronary angioplasty. AB - The purpose of this investigation was to evaluate the relation of coronary artery stenosis and associated pressure gradient to the magnitude of exercise-induced left ventricular dysfunction in patients with single vessel coronary artery disease. The percent stenosis and minimal cross-sectional area were measured before and after percutaneous transluminal coronary angioplasty and compared with radionuclide measurements of left ventricular function before and after angioplasty in 41 patients with proximal left anterior descending coronary artery lesions, providing 82 points of comparison. The gradient could be measured for 75 comparisons. Forty stenoses less than 50% were associated with a mean left ventricular exercise ejection fraction of 0.66 +/- 0.08 (mean +/- SD), 25 stenoses from 50 to 75% with a mean ejection fraction of 0.59 +/- 0.12 and 17 stenoses greater than 75% with a mean ejection fraction of 0.49 +/- 0.08. Thirty five stenoses with a gradient less than 20 mm Hg were associated with a mean ejection fraction of 0.65 +/- 0.09, 24 with a gradient from 20 to 50 mm Hg with a mean ejection fraction of 0.58 +/- 0.13 and 16 with a gradient greater than 50 mm Hg with a mean ejection fraction of 0.53 +/- 0.10. These data document a relation between the magnitude of coronary artery stenosis and associated gradient to exercise-induced left ventricular dysfunction in homogeneous patient groups. However, discordance of these variables occurs commonly in individual patients. PMID- 3036926 TI - Effect of wheat bran on bowel function and fecal calcium in older adults. AB - Seven healthy older volunteers participated in a 33-day study consisting of three sample collection periods, a 10-day control, and two 10-day experimental periods. Subjects consumed their usual self-selected diets throughout and a daily wheat bran supplement (30 g) during the two experimental periods. Food intake was recorded daily by subjects and accuracy and completeness checked daily by personal interview. Apparent calcium absorption decreased significantly from 22.1 +/- 5.6% (mean +/- SD) during the control to 8.6 +/- 5.2% during the second bran period. The wheat bran supplement significantly increased wet and dry stool weights but had no effect on stool moisture or defecation frequency. Gastrointestinal transit time of a dose of chromium decreased significantly, from 75 +/- 33 to 54 +/- 19 hr; of a dose of polyethylene glycol insignificantly, from 98 +/- 59 to 69 +/- 46 hr. Mean recovery of 21 doses of chromium of 98.7 +/- 5.0% verified that stool collection was complete. The results suggest that the ability of wheat bran to regulate bowel function in the apparently healthy older adult may be accompanied by increased fecal calcium losses similar to what has been reported for younger adults. PMID- 3036927 TI - Autoradiographic localization of substance P binding sites in guinea-pig airways. AB - The distribution of substance P (SP) binding sites in guinea-pig airway was examined by in vitro autoradiography with tritium- and iodine-labeled SP. Specific SP binding sites were most abundant in tracheobronchial smooth muscle but were also detected in the mucosa/submucosa. Binding within the mucosa/submucosa was especially high in the region of glands. Binding of iodine labeled SP to cartilage was negligible. Tritium-labeled SP bound non-specifically to airway cartilage. These observations are consistent with the proposed effects of SP-containing afferent nerves on airway resistance and vascular permeability. The localization of specific SP binding sites suggests that SP may also affect exocrine glands in the respiratory tract. PMID- 3036928 TI - Biochemical changes in the intestine associated with anoxia and reoxygenation: in vivo and in vitro studies. AB - In ischemia/reperfusion injury, it is hypothesized that superoxide is responsible for the component of injury due to reperfusion. The superoxide is hypothesized to result from the aerobic oxidation of purines produced by the ischemia-mediated breakdown of high-energy phosphates. This oxidation is catalyzed by xanthine oxidase proposed to be rapidly formed as a result of ischemia-mediated protease conversion from xanthine dehydrogenase. In vivo experiments with the intestine of either rats or guinea pigs were unable to confirm the rapid conversion of xanthine dehydrogenase to xanthine oxidase as a result of ischemia. In vitro experiments with isolated guinea pig enterocytes did show a significant increase in xanthine oxidase activity after these cells were first placed in an anaerobic environment for 60 min and then reoxygenated; however, the magnitude of the increase is such that the biological importance of this finding remains uncertain. Using a variety of techniques, including spin trapping, hydroxylamine oxidation, and vanadate NADPH oxidation, we explored the possibility that superoxide was produced as a result of anoxia followed by reoxygenation in the in vitro enterocyte system. From these experiments, we determined that superoxide is generated as a result of anoxia/reoxygenation. However, from xanthine oxidase inhibition experiments using pterinaldehyde, only a small percentage of the total superoxide produced comes from the action of this enzyme on purines. PMID- 3036929 TI - Approaches to prevention of asbestos-induced lung disease using polyethylene glycol (PEG)-conjugated catalase. AB - Asbestos-associated damage to cells of the respiratory tract in vitro can be prevented by the simultaneous addition of scavengers of active oxygen species to cultures. To determine if administration of scavenger enzymes to animals and humans is a plausible approach to the prevention of asbestos-induced lung disease, osmotic pumps were filled with various concentrations of PEG-coupled catalase and implanted subcutaneously into Fischer 344 rats over a 28-day period. At 3, 14, and 28 days after implantation of the pumps, the animals were evaluated for levels of catalase in serum and lung. In addition, lung tissue and lavage fluids were examined at 28 days for biochemical and morphologic indications of cell injury, inflammation, and fibrotic lung disease. At all time points examined, the administration of PEG-catalase caused a dosage-dependent increase in serum levels of catalase. The levels of lung catalase were evaluated at 28 days but not at earlier time periods. In comparison to control rats, the amounts of enzymes (lactic dehydrogenase, alkaline phosphatase), protein, and cells in lavage fluids from treated animals were unaltered. Moreover, the lungs showed no evidence of inflammation or fibrotic disease as determined by differential cell counts in lavage and measurement of hydroxyproline. These studies suggest that administration of PEG-catalase does not cause injury or other alterations in lung tissue and can be pursued as a feasible approach to prevention of asbestosis. PMID- 3036930 TI - Purity of catalase preparations: contamination by endotoxin and its role in the inhibition of airway inflammation. PMID- 3036931 TI - Diet restriction retards the age-related loss of beta-adrenergic receptors and adenylate cyclase activity in rat lung. AB - In the rat, dietary restriction prolongs life span and retards a variety of physiological processes that change with age. Beta-adrenergic responsiveness declines with age. We assessed beta-adrenergic receptor density and adenylate cyclase activity in lungs of 6-, 18-, 24-, and 27-month-old rats fed ad libitum or restricted to 60% of the ad libitum food after 6 weeks of age. Beta-adrenergic receptor density declined in the ad libitum group (M +/- SE, 417 +/- 30, 349 +/- 23, 297 +/- 19, 260 +/- 30 fmol/mg protein, p less than .01). In the diet restricted group, beta-adrenergic receptor density declined between 6 and 18 months (493 +/- 35, 370 +/- 28, p less than .05) but remained unchanged from 18 to 27 months (370 +/- 28, 333 +/- 19, 360 +/- 16). Isoproterenol-stimulated adenylate cyclase activity declined with age in the ad libitum group (53.4 +/- 7.5, 46.8 +/- 8.8, 38 +/- 3.9, 27.8 +/- 3.3 p mol cAMP/mg protein, p less than .05) but not in diet-restricted group (45.1 +/- 8.9, 44 +/- 7.9, 41.8 +/- 2.4, 47.3 +/- 5.1). Changes in forskolin-stimulated adenylate cyclase activity with age were not affected by diet restriction. These data suggest that diet restriction retards some of the age-related changes in the beta-adrenergic pathway. PMID- 3036932 TI - Nerve conduction studies in infants and children. AB - The electrophysiologic evaluation of peripheral nerves may provide critically important information, both with respect to diagnosis and prognosis, in the child with a suspected neuromuscular disorder. However, special attention to various technical considerations is necessary to avoid misleading results. Utilizing these techniques, both hereditary and acquired neuropathies may be identified and characterized. The latter has become especially important in view of recent advances in the treatment of acquired demyelinating neuropathies. PMID- 3036933 TI - Suppressed pituitary ACTH response after ACTH treatment of infantile spasms. AB - Suppression of an adrenocorticotropic hormone (ACTH) response to insulin hypoglycemia has been reported in ACTH-treated adults. There are no guidelines for withdrawal of ACTH treatment in children. After observing suppressed morning cortisol in several children, insulin tolerance tests were performed in five children within 48 hours after tapered withdrawal of ACTH treatment for myoclonic seizures. ACTH response, as determined by cortisol and beta-endorphin radioimmunoassay, was adequate in four of the children. One child showed low basal levels and minimal elevation during hypoglycemia for both beta-endorphin (0 to 3 pg/ml) and cortisol (3.6 to 4.4 micrograms/dL) on initial testing, but normal responses six weeks later. Measurement of beta-endorphin response supported a central basis for suppression in the child, who had had an adrenal hemorrhage during gram-negative sepsis while on ACTH. ACTH release is transiently suppressed in some children after exogenous ACTH treatment. Tapered withdrawal and stress coverage is recommended. PMID- 3036934 TI - Continuous infantile spasms as a form of status epilepticus. AB - An infant with congenital cytomegalovirus infection first developed seizures at six weeks of age. At 3 1/2 months of age, he developed continuous infantile spasms that lasted for more than an hour. This episode of status epilepticus was terminated by intravenous lorazepam and paraldehyde, and seizures were subsequently controlled for seven months by adrenocorticotropic hormone (ACTH), valproic acid, and phenobarbital. This case demonstrates that continuous infantile spasms may occur as a unique form of status epilepticus in young infants. PMID- 3036935 TI - Electromyography in infants and children. AB - Electromyography (EMG) is of proven value in the diagnosis of acute and chronic neuromuscular diseases in infants and children. When this technique is combined with nerve conduction studies, including repetitive nerve stimulation studies, it is often possible upon completion of the studies to identify the disorder as one of nerve, neuromuscular junction, or muscle. The purpose of this article is to review the principles and techniques of EMG in infants and children and to describe the EMG findings in several neuromuscular disorders. PMID- 3036936 TI - Interferon therapy of chronic hepatitis B virus infection in Chinese. AB - Hepatitis B virus (HBV) infection is prevalent in Chinese populations. 40.4% of 383 Chinese HBV carriers studied were HBeAg-positive. The annual rate of spontaneous clearance of HBeAg was 11%. Twenty-six patients with HBsAg- and HBeAg and HBeAg-positive non-malignant chronic liver disease randomised to receive recombinant alpha-2 interferon or no treatment have been followed for 6 months or longer. Seven of the 20 treated patients cleared HBeAg during or shortly after treatment but this was sustained in only 1 patient. One of the 6 controls had transient loss of HBeAg. It is too early to conclude whether interferon has any long-term effect on the suppression of HBV replication in Chinese patients. Sixty nine patients with histologically proven hepatocellular carcinoma were randomised to receive adriamycin or interferon. Although there was no significant benefit on survival, interferon therapy was associated with greater than 25% regression in tumor size in 12.5% of patients and less toxicity. PMID- 3036937 TI - Conjugates between monoclonal antibodies to HBsAg and cytosine arabinoside. AB - Chemotherapeutic agents such as cytosine arabinoside (ARA-C) and adenine arabinoside (ARA-A) have been shown to eliminate or suppress replication of some DNA viruses. These effects were however associated with systemic side-effects to the treated hosts. We are currently exploring a strategy to construct conjugates between these antiviral agents and monoclonal antibodies directed against viral surface proteins. Such conjugates should enable specific delivery of the antiviral agents to their preferred site of action. In the present study, monoclonal antibodies to hepatitis B virus surface antigen (HBsAg) of the IgG2a and IgM isotypes were conjugated to ARA-C via a dextran bridge. The use of dextran enables the binding of a high number of drug molecules to the antibody with minimal loss of activity. Briefly, partially oxidized dextran was coupled to ARA-C and then to monoclonal anti-HBs. Following the conjugation process, the IgG2a anti-HBs-(dex)-ARA-C conjugate retained its capacity to bind to HBsAg fixed to a solid matrix as compared to non-conjugated homologous antibodies. Conjugates between ARA-C and IgM anti-HBs lost a significant degree of their binding activity to purified HBsAg. However, conjugates containing anti-HBs of both isotypes bound specifically to PLC/PRF/5 human hepatoma cells that express HBsAg on their cell surface. Conjugates containing non relevant monoclonal antibodies did not bind to target cells. Pharmacologic activity of the various compounds was assessed by an [3H]thymidine incorporation assay in hepatoma cells in culture. IgM and IgG2a containing conjugates caused suppression of [3H]thymidine incorporation into PLC/PRF/5 cells. This effect was more pronounced for conjugates containing monoclonal IgM anti-HBs.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3036938 TI - Vitamin E deficiency and its clinical significance in adults with primary biliary cirrhosis and other forms of chronic liver disease. AB - The vitamin E status of 146 adults with chronic liver disease was assessed by estimating both their serum vitamin E concentration and the ratio of serum vitamin E to serum cholesterol concentration. Low levels of vitamin E occurred most frequently in patients with primary biliary cirrhosis and other forms of chronic cholestatic liver disease. When a serum vitamin E concentration of 12.3 mumol/l (mean-2 SD of a control population) was taken as the lower limit of normal, 44% of patients with primary biliary cirrhosis and 32% with other chronic cholestatic liver disease had a reduced concentration, indicating a biochemical deficiency of vitamin E. If a vitamin E/total cholesterol ratio of 2.35 mumol/mmol was taken as the lower limit of normal, then 64% and 43% of patients with primary biliary cirrhosis and other chronic cholestatic liver disease, respectively, had a biochemical deficiency of vitamin E. Of the patients with chronic cholestasis and a serum bilirubin concentration greater than 100 mumol/l, 91% had a reduced vitamin E/cholesterol ratio. Twelve patients with primary biliary cirrhosis and severe vitamin E deficiency (serum vitamin E less than 5.0 mumol/l and a vitamin E/cholesterol ratio less than 1.0 mumol/mmol) underwent extensive neurological investigation. Five had a mild mixed sensorimotor peripheral neuropathy, which was not, however, typical of the neurological syndrome associated with vitamin E deficiency. In patients with severe biochemical deficiency of vitamin E (less than 5 mumol/l and less than 1 mumol/mmol total cholesterol), administration of large oral doses of vitamin E only increased serum concentrations to within the normal range in one patient; in the others even weekly parenteral administration over a 3-month period did not correct deficiency. In patients with less severe biochemical deficiency, the serum vitamin E concentration and vitamin E/total cholesterol ratio were restored to normal by oral or intramuscular supplements of the vitamin. PMID- 3036939 TI - What is the effect of a cytomegalovirus infection when superimposed on HBsAg positive chronic hepatitis? AB - A non-transfusion-related cytomegalovirus infection was observed in two patients with chronic HBsAg-positive liver disease. Both patients were immunodepressed and died of acute hepatic failure. A superimposed CMV infection has a high fatality rate in the case of underlying chronic type B liver disease. PMID- 3036940 TI - Production of abnormal prothrombin (des-gamma-carboxy prothrombin) by hepatocellular carcinoma. A clinical and experimental study. AB - We measured plasma abnormal prothrombin (des-gamma-carboxy prothrombin; DCP) levels in normal subjects and in patients with hepatocellular carcinoma and other various diseases using the enzyme-linked immunosorbent assay developed by Motohara et al. (Pediatr Res 1985; 19: 354-357). Fifty-eight percent of 52 patients with hepatocellular carcinoma had elevated DCP levels; 24 of 28 patients with advanced or moderately advanced hepatocellular carcinoma were positive. By contrast, 50 normal controls, 13 pregnant women and 10 patients with acute hepatitis had normal levels. Three of 55 patients with chronic liver disease, and 6 of 32 patients with other malignancies, showed a slight increase. Thus, increased plasma DCP appears useful for the diagnosis of hepatocellular carcinoma. To elucidate the mechanism for the increase of DCP in hepatocellular carcinoma, we cultured a human hepatoma cell line, huH-2, and measured the levels of this abnormal prothrombin in the medium. The huH-2 cells produced large amounts of DCP in the medium without added vitamin K. It increased in a cell concentration- and time-dependent fashion. These cells produced no detectable amount of DCP in the medium with added vitamin K. Thus, human hepatoma cell line huH-2 produces DCP, and its production is dependent on the amount of vitamin K available in the medium. Des-gamma-carboxy prothrombin may be a useful tumor marker for the diagnosis of hepatocellular carcinoma. PMID- 3036941 TI - Benzodiazepines, plasma MHPG and alpha-2-adrenoceptor function in man. AB - The effects of a single (15 mg) dose of diazepam, a 3-week course of diazepam (25 mg/d) and its withdrawal on plasma MHPG and the reduction produced by clonidine (1.5 micrograms kg-1 i.v.) were studied in 8 male volunteers. Chronic diagram reduced, but not significantly, plasma MHPG and attenuated the reduction produced Gyclonidine. During the withdrawal phase MHPG levels were significantly enhanced and the lowering produced by clonidine was also greater. These results support the hypothesis that some aspects of benzodiazepine withdrawal may reflect excessive central noradrenergic activity. PMID- 3036942 TI - Quantitative immunoelectron microscopic localization of (Na+,K+)ATPase in rat parotid gland. AB - Distribution of (Na+,K+)ATPase on the cell membranes of acinar and duct cells of rat parotid gland was investigated quantitatively by immunoelectron microscopy using the post-embedding protein A-gold technique. In acinar cells, ATPase was localized predominantly on the basolateral plasma membranes. A small but significant amount of (Na+,K+)ATPase was, however, detected on the luminal plasma membranes, especially on the microvillar region of the acinar cells; the surface density on the luminal membrane was approximately one third of that on the basolateral membranes. In duct cells, many gold particles were found on the basolateral membrane, especially along the basal infoldings of the plasma membranes, whereas no significant gold particles were found on the luminal plasma membranes, suggesting unilateral distribution of ATPase in duct cells. We suggest that in acinar cells sodium ion is not only transported paracellularly but is also actively transported intracellularly into the luminal space by the (Na+,K+)ATPase located on the luminal plasma membranes, and that water is passively transported to the luminal space to form a plasma-like isotonic primary saliva, while in the duct cells the same ion is selectively re-absorbed intracellularly by (Na+,K+)ATPase found in abundance along the many infoldings of the basal plasma membranes, thus producing the hypotonic saliva. PMID- 3036943 TI - Identification of interferon-gamma as the lymphokine that mediates leukotriene B4 induced immunoregulation. AB - Leukotriene B4 (LTB4) has been shown to modulate lymphocyte responses in both a positive and a negative way, depending on the particular cell subsets it interacts with. Recent evidence also indicates that LTB4 can directly affect the production of cytokines such as interleukin 1 (IL 1) or interleukin 2 (IL 2) and interferon-gamma (IFN-gamma). In this report, we present evidence that human T cells pulsed with LTB4 modulate IL 1 production by human monocytes by secreting IFN-gamma. In fact, we found that LTB4-pulsed T cells were capable of inducing a suppression of lymphocyte proliferation if allowed to interact with monocytes, but that this suppression was reversed to an enhancing effect when monocytes were treated with the cyclooxygenase inhibitor indomethacin. Furthermore, LTB4-pulsed T cells released a soluble factor that would mediate both effects. This factor was found to be IFN-gamma, because affinity-purified IFN-gamma could reproduce the effects, and a rabbit polyclonal anti-serum to human IFN-gamma could block the activities of supernatants from LTB4-pulsed T cells. LTB4 was also shown to enhance IFN-gamma production by T4+ T cells and to inhibit IFN-gamma production by T8+ T cells. These results suggest that LTB4 may regulate immune cell functions by inducing IFN-gamma production by T4+ cells. PMID- 3036944 TI - Effects of bacterial lipopolysaccharide on the hydrolysis of phosphatidylinositol 4,5-bisphosphate in murine peritoneal macrophages. AB - LPS and lipid A initiated enhanced hydrolysis of PIP2 in macrophages. When murine peritoneal macrophages were labeled with [2-3H]myoinositol and stimulated with either LPS or lipid A, a rapid (within 10 sec) rise in Ins(1,4,5)P3 was observed. The breakdown pattern of Ins(1,4,5)P3 was complex; this included breakdown of Ins(1,4,5)P3 and formation of Ins(1,3,4,5)P4 (approximately 10 to 30 sec), and ultimately formation of Ins(1,3,4)P3 (approximately 60 sec). Within 10 sec after treatment, LPS caused an average increase of about fourfold to fivefold in Ins(1,4,5)P3, which declined over 5 min. When the total isomers of InsP3 were measured, levels rose about twofold in response to LPS or to lipid A and remained elevated for as long as 5 min. Lipid A, in the concentration range of 0.1 to 10 micrograms/ml, induced elevated intracellular levels of Ca2+ as quantified by fluorescence with Quin 2 or with Fura 2. When single, adherent Fura 2-loaded macrophages were treated with lipid A, basal levels of calcium rose over 10 sec from approximately 55 nM to almost 600 nM. LPS, paradoxically, did not cause such substantial increases in intracellular calcium (i.e., increases of approximately 26 nM) when judged by Fura 2 fluorescence. LPS treatment led to enhanced phosphorylation of a characteristic set of proteins, similar to those induced by stimulating protein kinase C (PKC) with phorbol myristate acetate as previously reported. The enhanced phosphorylation of pp28, pp33, and pp67 in macrophages was evident by 15 min and optimal by 30 min. Taken together, these observations indicate that LPS and lipid A cause increased breakdown of phosphatidylinositol 4,5-bisphosphate, which led to enhanced intracellular levels of calcium and also to enhanced protein phosphorylation, presumably mediated by PKC. The data thus suggest that one major intracellular signal transduction mechanism, initiated by LPS and lipid A in macrophages, is the rapid breakdown of PIP2. PMID- 3036945 TI - Recognition of EBV plasma membrane protein expressed on murine cells after gene transfer. AB - The immune response to B lymphocytes infected with Epstein-Barr virus (EBV) prevents their overgrowth in normal humans. A murine model is now described for analyzing the T cell immune response to Epstein-Barr virus genes expressed in murine lymphoblasts by gene transfer. In mice, a 60,000 dalton virus-encoded protein characteristically found in the plasma membrane of latently infected human lymphocytes readily induces both proliferative and cytolytic T lymphocytes specific for both the EBV protein and murine major histocompatibility proteins. Longterm cultures of L3T4+ cells, some of which were cytolytic, were found to be restricted by H-2I-Ed and the latent membrane protein. Similarly, Lyt-2+ cells were cytolytic and were restricted by H-2Ld and the lymphocyte membrane protein gene product. The similarity in murine and human effector cell responses suggests that this is a useful experimental model, and the EBV latent infection membrane protein may be an important antigen in the immune restriction of growth transformed latently infected lymphocytes. PMID- 3036946 TI - Structure-function relationships for the IL 2-receptor system. IV. Analysis of the sequence and ligand-binding properties of soluble Tac protein. AB - The Tac protein is one of at least two glycoproteins known to bind the growth and differentiation factor interleukin 2 (IL 2). In addition to its location on the cell surface, where it plays a part in high and low affinity IL 2 receptors, Tac is released from activated lymphocytes in a soluble form. We observed this release both for Tac protein labeled biosynthetically and for Tac protein labeled by surface iodination of intact cells. Competitive binding studies indicated that the soluble Tac protein retained an ability to bind IL 2 with a low affinity (Kd of 11.1 nM). In addition, structural analysis revealed that the polypeptide chain began at position 1 and ended at or just before Cys-192 of the full-length molecule. Thus, the protein was missing its normal transmembrane and intracytoplasmic segments, accounting for its solubility and cellular release. The apparent lack of modification in the amino acid sequence and the termination at Cys-192 are inconsistent with a mechanism of cellular release dependent only on alternate mRNA splicing. Instead, the results suggest that proteolysis may accompany the release of soluble Tac protein from cells expressing IL 2 receptors. PMID- 3036947 TI - Murine natural killer cells limit coxsackievirus B3 replication. AB - Previous indirect evidence suggested that natural killer (NK) cells play a role in coxsackie virus B3 serotype 3, myocarditic variant (CVB3m)-induced myocarditis by limiting virus replication. In this study, we present direct evidence that NK cells can limit CVB3m replication both in vitro and in vivo. Virus titers are lowered in primary murine neonatal skin fibroblast (MNSF) cultures incubated with activated splenic large granular lymphocytes (LGL) taken from mice 3 days postinoculation of CVB3m, a time of maximal NK cell activity. The antiviral effect of this cell population is diminished by complement-mediated lysis with the use of anti-asialo GM1 antiserum but not with anti-Lyt-2 monoclonal antibody. Neither interferon nor anti-CVB3m-neutralizing antibody was detected in these cultures. Although activated LGL initiate lysis within CVB3m-infected MNSF in vitro within 3 hr of addition, they do not lyse uninfected MNSF cultures. CVB3m replication is required for expression of surface changes on MNSF that result in lysis by NK cells because cell cultures treated with compounds that prevent CVB3m replication are not killed by LGL. LGL also do not lyse MNSF cultures inoculated with UV-inactivated virus. Mice inoculated with activated LGL and subsequently challenged with CVB3m had reduced titers of virus in heart tissues in comparison to titers of CVB3m in heart tissues of mice not given LGL. The antiviral activity of the LGL preparation was abolished by prior treatment with anti-asialo GM1 antiserum plus complement but not by prior treatment with anti-Lyt-2 monoclonal antibody and complement. These data suggest that NK cells can specifically limit a nonenveloped virus infection by killing virus-infected cells. PMID- 3036948 TI - B lymphocyte activation. Transmembrane signal transduction by membrane immunoglobulin in isolated cell membranes. AB - We report the development of cell-free systems in which ligation of B cell membrane immunoglobulin leads to demonstrable mono- and polyphosphatidylinositol hydrolysis. Membranes were prepared by differential centrifugation of sonicates of normal murine B cells. Incubation of these membranes with 32P-adenosine triphosphate in the presence of Mg2+ effected the radiolabeling of phosphatidylinositol 4,5-bisphosphate (PtdInsP2) and phosphatidylinositol 4 phosphate (PtdInsP). Alternately, membranes were labeled with exogenous 3H(inositol)-PtdInsP2 in sodium cholate. Stimulation of labeled membranes with anti-immunoglobulin, but not anti-Ia or anti-H2 antibodies, resulted in hydrolysis of phosphatidylinositol, PdtInsP, PtdInsP2 and generation of inositol phosphates indicative of activation of a phospholipase C. The response was rapid, being detectable within 30 sec of stimulation, and independent of Ca2+ and guanosine 5'-triphosphate. Optimal responses were dependent on the presence of a cytosolic factor presumed to be phospholipase C. Development of these systems represents an important step towards reconstitution of membrane immunoglobulin mediated transmembrane signaling in artificial membranes. PMID- 3036949 TI - Mouse lymphocyte enzymatic markers. A rapid method for achieving selective solubilization and efficient recovery of the membrane-bound 5'-nucleotidase. AB - A simple method is described for achieving a good recovery and a partial purification of the membrane-bound 5'-nucleotidase (5'-N) from mouse lymphocytes. The experimental procedure is based upon plasma membrane isolation on polycationic beads and selective solubilization of the enzyme activity from bead bound plasma membranes. With this method, more than 95% of the 5'-N activity detectable in the whole cell homogenates can be routinely recovered in a single fraction showing a 5'-N specific activity which is at least 60 times higher than that found in the crude homogenate. This method also provides a complete separation of 5'-N from the membrane-bound alkaline phosphatase (AP), as well as from any other interfering non-specific phosphatase. Since this method is rapid and highly reproducible even when small amounts of lymphocytes are available, it may be useful for detecting changes in 5'-N activity in the different T- and B lymphocyte subpopulations. PMID- 3036950 TI - Rapid simultaneous assessment of neutrophil superoxide generation and lysosomal enzyme release. AB - A rapid method for the simultaneous measurement of neutrophil superoxide generation and beta-glucuronidase release is described. Assay of beta glucuronidase using a fluorescent substrate is shown to be valid in the presence of reduced or unreduced ferricytochrome C, a prerequisite for the simultaneous assessment of this enzyme activity and O-2 generation. PMID- 3036951 TI - Antibody reactivity to a synthetic peptide (P62) of the Epstein-Barr nuclear antigen in sera of patients with X-linked lymphoproliferative syndrome. AB - An enzyme-linked immunosorbent assay (ELISA) was used to measure IgG antibody titers against a synthetic peptide whose sequence was derived from the glycine alanine repeating region of Epstein-Barr virus nuclear associated antigen 1 (EBNA 1). Antibody titers were determined in sera from 15 normal subjects, sera from 21 normal male siblings of X-linked lymphoproliferative syndrome (XLP) patients, from 20 XLP patients comprising a total of 42 samples, and ten samples before and ten samples after gamma-globulin therapy in ten patients with XLP. Data analysis demonstrated that while there are differences between the ELISA and ACIF, they appear to measure a similar response as demonstrated by their correlation coefficient (0.77) and the GMT to EBNA observed by both methods. No cross reactivity of cytomegalovirus antibodies to the EBNA-1 peptide was observed by immunoblotting, ELISA or ACIF using adsorption against AD-169 infected MRC-5 cells. However, non-specific binding was observed if samples were not pre incubated in a 10% goat serum PBS-Tween 20 solution. This pre-treatment removed the non-specific binding that falsely elevated the GMT in approximately 15% of both normal and XLP samples in ELISA. The ELISA system appears to be a sensitive, reproducible and objective test that may be useful for assessing the antibody response of patients to the EBNA-1 protein. PMID- 3036953 TI - Risk of transmitting the human immunodeficiency virus, cytomegalovirus, and hepatitis B virus to health care workers exposed to patients with AIDS and AIDS related conditions. AB - This prospective cohort study was designed to evaluate the risk of occupational transmission of human immunodeficiency virus (HIV), hepatitis B virus (HBV), and cytomegalovirus (CMV) to health care workers with intensive exposure to HIV infected patients. Seventy-five percent of the 270 subjects had been exposed to patients with AIDS and AIDS-related conditions (ARC) for at least one year before enrollment, 18% worked in specialized AIDS units, and 35% sustained a total of 342 accidental parenteral exposures to HIV-infected body fluids. At the time of enrollment, none had antibody to HIV, and none of the 175 subjects retested 10 months later had acquired antibody. No evidence of increased risk of acquiring CMV or HBV was obtained. These results indicate that health care workers are at minimal risk for HIV, CMV, and HBV transmission from occupational exposure to patients with AIDS or ARC, even when intensively exposed for prolonged periods of time. PMID- 3036954 TI - Construction of defined galE mutants of Salmonella for use as vaccines. AB - We describe the molecular cloning of the gal operon of Salmonella typhimurium LT2 and the localization of the gal promoter and the genes galE, galT, and galK. The order of the genes and the direction of transcription was the same as in Escherichia coli K12. A restriction enzyme map of the operon was obtained, and the approximate termini of the gal genes were located by using transposon insertion mutagenesis and minicell analysis. We constructed a plasmid that contained a defined 0.4-kilobase deletion in the galE but still expressed galT and galK activities from the gal promoter. This galE deletion was recombined into the chromosomal gal operons of S. typhimurium and Salmonella typhi Ty2. The resulting strains were vigorous, nonreverting galE mutants that were sensitive to galactose-induced lysis at 0.2 mM galactose. The S. typhimurium galE derivatives were avirulent and protective in mice. PMID- 3036955 TI - Human parvovirus B19 infection during pregnancy. AB - Human parvovirus B19 (B19) has been implicated as the cause of fifth disease and has been associated with fetal death. We identified pregnant women who were at risk of contracting B19 infection during an outbreak of fifth disease. The sera of 12 women classified at high risk of exposure and 19 classified at low risk were tested during prenatal care, at delivery, or at both times for IgG and IgM antibodies to B19. Four women at high risk but none at low risk were considered infected because they were IgM positive. One IgM-positive woman gave birth to a stillborn hydropic fetus whose tissues were positive for B19 DNA by a nucleic acid hybridization assay. The other three IgM-positive women gave birth to normal offspring, of whom one had IgM-positive cord serum. We conclude that B19 infection during pregnancy can lead to fetal infection with at least two associated outcomes--no adverse effect on the fetus or fetal death. PMID- 3036952 TI - A rapid method for detection of cellular proliferation using carboxyfluorescein. Assay of growth factors (IL-2, IL-1) and growth inhibiting antibodies. AB - The uptake of carboxyfluorescein diacetate (CFDA) into live cells was used as the basis for a simple, rapid and fully automated micromethod for determination of cell growth. The aim of the investigation was to adapt the CFDA method for detection of cell growth factors from cell culture supernatants. Thus, the biological activities of the growth factors IL-2 or IL-1 could be detected and quantitated at the same level of sensitivity as with the conventional [3H]thymidine incorporation. Similarly, the method was well suited to assay inhibition of IL-2-dependent lymphocyte growth by the monoclonal antibody anti Tac, binding to the human lymphocyte membrane receptor for IL-2. The CFDA method proved to be rapid, reliable and well suited for several applications involving limiting numbers of cells and biological reagents. PMID- 3036956 TI - Lack of association of cytomegalovirus with endemic African Kaposi's sarcoma. PMID- 3036957 TI - Role of antigen-presenting cells in congenital cytomegalovirus-specific immunodeficiency. PMID- 3036958 TI - Serological markers for Epstein-Barr virus infection in chronic carriers of hepatitis B surface antigen who live in northern Canada. PMID- 3036959 TI - Perinatal transmission of coxsackievirus B3 in mice. AB - Oral infection of pregnant mice with coxsackievirus B3 (CB3) late in gestation produced maternal viremia, which peaked three to four days after challenge, then rapidly diminished. Ninety percent of mice developed IgG antibody to CB3 by ten days after challenge. CB3 titers from placental tissue peaked at two to four days after maternal infection, but moderate titers persisted for at least eight days. Though virus could be recovered (less than 1.0-1.5 log10 pfu/g) from fetal tissue in only a small percentage (3%-13%) of pregnancies, virus could be found in some fetuses eight days after maternal infection. Despite the low rate of fetal infection, many pups born to dams infected one to eight days before delivery were either stillborn (4%-41%) or died from infection shortly after birth (14%-68%). The actual percentages depended on the time interval from infection of the dams to delivery. Maternal IgG antibody to CB3, which appeared at six to eight days after infection, protected against postnatal mortality, but did not protect against stillbirth. PMID- 3036960 TI - Factors associated with the pathogenesis of AIDS. PMID- 3036961 TI - Selective lack of antibody to a component of EB nuclear antigen in patients with chronic active Epstein-Barr virus infection. AB - The sera of 12 patients with presumed chronic active Epstein-Barr virus (EBV) infection lacked antibody to a component of the Epstein-Barr nuclear antigen (EBNA) complex encoded by the BamHI K fragment of viral DNA. This anomaly, detected in approximately 18% of sera obtained from patients with a diagnosis of "chronic mononucleosis," was more often found in patients with severe disease (approximately 32%) who had objective clinical findings and markedly elevated antibody titers to EBV replicative antigens than in those patients with the "fatigue syndrome" (10%). The lack of antibody to the K nuclear antigen is specific because most of those who did not have antibody to the K antigen made antibody to other latent nuclear (EBNA 2) antigens or nuclear early antigens. Such patients are thus able to lyse immortalized cells, release nuclear products, and present them to the immune system. Three hypotheses are suggested to explain the lack of antibody to the K antigen: a viral mutation, a failure of immune recognition, or lack of in vivo expression of the antigen due to extensive viral replication. Lack of antibody to one component of EBNA may serve as an objective serological marker for certain patients with chronic EBV infection. PMID- 3036962 TI - Rhinovirus infections in Tecumseh, Michigan: frequency of illness and number of serotypes. AB - We studied rhinovirus-associated illnesses from 1976 to 1981 among residents of the community of Tecumseh, Michigan, in a continuation of similar studies done in 1966-1971. Rhinoviruses were the most frequently isolated respiratory pathogen in all age groups, including young children. The pattern of age-specific isolation rates was similar to that for total respiratory illness. Rhinovirus-associated illnesses were generally mild but of relatively long duration; restriction of daily activity was frequent, especially in certain age groups. Typing of all isolates has been completed; with high-quality antisera nearly all isolates could be typed. Thus, new serotypes are probably not evolving, and most have already been identified. Differences in frequency of occurrences of different serotypes were evident, but ranking in order of importance was difficult based on existing data. PMID- 3036963 TI - Nosocomial cytomegalovirus infections within two hospitals caring for infants and children. AB - Using serology, virology, and molecular epidemiology, we investigated nosocomial transmission of cytomegalovirus (CMV) over a two-year period in two contrasting environments: a crowded, busy pediatric chronic care unit (337 patients, 43 nurses, and 76 therapists; average prevalence of CMV excretion in patients, 16%) and a small neonatal unit (293 patients and 69 nurses; average prevalence, 0.7%). In the chronic care unit no nurse or therapist acquired CMV, but two pairs of infants were infected with homologous strains of CMV, and patient-to-patient transmission was proven in one pair. In the neonatal unit no patients acquired CMV in the hospital, but two nurses seroconverted, with a nonoccupational source proven for one. Transmission from CMV-infected caretaker to patient did not occur in either environment. CMV was isolated from diapers as well as hands of patients and personnel but not from other environmental surfaces. PMID- 3036964 TI - Epstein-Barr virus, cytomegalovirus, and other viral infections in children after liver transplantation. AB - We studied 51 consecutive pediatric patients for the frequency and morbidity of viral infections after liver transplantation. The incidence of primary (67%) and reactivation (48%) Epstein-Barr virus (EBV) infections and reactivation (88%) cytomegalovirus (CMV) infection was comparable to that seen in adult transplant recipients. However, fewer pediatric than adult transplant recipients experienced primary CMV infection (P less than .01). Five (38%) of 13 CMV infections were symptomatic and included hepatitis, pneumonitis, enteritis, and mononucleosis. Two of 14 patients with primary EBV infection subsequently developed, at two months and two years after initial infection, an EBV-associated lymphoproliferative syndrome, and one of 10 patients with reactivated EBV infection developed a possible EBV-associated febrile encephalopathy. Other viruses causing infection in these children included herpes simplex virus, varicella-zoster virus, adenovirus, parainfluenza virus, respiratory syncytial virus, and rotavirus. PMID- 3036965 TI - A comparative study of herpes simplex infections in renal transplant and leukemic patients. AB - Herpes simplex virus (HSV) reactivation and lesion formation were studied in 68 renal transplant recipients and 30 leukemic patients. Antibody titers to HSV were determined, and seropositive patients were examined three times weekly for up to one month. Surveillance cultures were taken for oral HSV, and HSV culture and cytology were done for all oral lesions found. In a smaller number of patients, immune responses were determined. HSV reactivation was similar in the transplant and leukemic groups (46.8% vs. 50%), but a significant difference in the incidence of HSV lesion formation was noted between the two groups. Of the transplant patients in whom HSV reactivated, 31.8% developed HSV lesions; of leukemic patients in whom HSV reactivated, 100% developed HSV lesions. Differences in the incidence of formation of HSV lesions in these groups of immunosuppressed patients suggest that reactivation of HSV and formation of HSV lesions may involve different mechanisms. Low levels of antibody-dependent cellular cytotoxicity were noted in leukemic patients and may contribute to increased formation of HSV lesions in this group. PMID- 3036966 TI - Reemergence of an epidemic coxsackievirus B5 genotype. AB - Outbreaks of coxsackievirus B5 (CB5) infections occur primarily during peak epidemic years, with comparatively few cases occurring during intervening years. This pattern of periodic CB5 epidemicity is quite distinct from the general endemicity typical of other group B coxsackieviruses. To determine the genetic relationships among CB5 isolates from different outbreaks, we compared viral RNAs by ribonuclease T1 oligonucleotide fingerprinting. Isolates obtained within an epidemic year had very similar fingerprints, an observation indicating that they were closely related variants of a single genotype. CB5 isolates from the major 1972 epidemic were not closely related to the genotype associated with the preceding epidemic of 1967. However, isolates from the most recent CB5 epidemic year, 1983, had fingerprints nearly identical to those of the 1967 strains. These findings provide clear evidence for epidemic reemergence of the 1967 genotype and suggest that the virus was maintained under conditions approaching evolutionary stasis during the intervening 16-year period. PMID- 3036968 TI - Minimal infectious dose of rotavirus in volunteers: safety issues. PMID- 3036967 TI - Human monkeypox: clinical features of 282 patients. AB - We present the clinical features and course of 282 patients with human monkeypox in Zaire during 1980-1985. The ages of the patients ranged from one month to 69 years; 90% were less than 15 years of age. The clinical picture was similar to that of the ordinary and modified forms of smallpox. Lymphadenopathy, occurring in the early stage of the illness, was the most important sign differentiating human monkeypox from smallpox and chickenpox. The symptoms, signs, and the course of the disease in patients who had been vaccinated against smallpox differed significantly from those in unvaccinated subjects. Pleomorphism and "cropping" similar to that in chickenpox occurred in 31% of vaccinated and 18% of unvaccinated patients. The prognosis depended largely on the presence of severe complications. No deaths occurred among vaccinated patients. In unvaccinated patients the crude case-fatality rate was 11% but was higher among the youngest children (15%). PMID- 3036969 TI - Multiple granular cell tumors of the palate. AB - A report of a case of bilateral granular cell tumors of the soft palate is presented. This is an unusual location for the tumor and is the first report of bilateral tumors at this site. PMID- 3036970 TI - "Spill" preventing measure by the use of hydroxylapatite. PMID- 3036971 TI - Hyperthermia-induced inhibition of respiration and mitochondrial protein denaturation in CHL cells. AB - Respiration of Chinese hamster lung V79 cells, as assayed by O2 consumption, increases linearly from 8 to 40 degrees C when plotted in the Arrhenius fashion but is strongly inhibited above 40 degrees C. The protein of mitochondria isolated from V79 cells undergoes structural transitions at 28 and 40 degrees C. This is supported by changes in the fluorescence excitation spectrum of conjugated pyrene maleimide and, to a lesser extent, intrinsic protein fluorophores. Electron spin resonance labelling studies with a derivative of tempo maleimide imply that extensive protein unfolding coincides with the 40 degrees C transition. The structural transition at 40 degrees C correlates well with inhibition of O2 consumption, is irreversible and is probably due to protein denaturation, while the change at 28 degrees C is reversible and has no effect on O2 consumption. Previous studies indicate the presence of a broad lipid transition extending from approximately 8 to 30 degrees C in mitochondrial membranes with all lipids being in the liquid-crystalline state above 30 degrees C. Thus, the onset of the lipid transition may induce the observed protein conformational change at 28 degrees C, but inhibition of respiration above 40 degrees C can be explained by protein denaturation alone. The region from 28 to 40 degrees C of stable protein conformation corresponds to the temperature range of V79 cell growth. PMID- 3036972 TI - Neuroendocrine changes in patients undergoing whole body hyperthermia. AB - A phase I study of whole body hyperthermia (WBH) (52 treatments/12 patients) utilizing a radiant heat device has been completed. This study incorporated a temperature escalation scheme from 39.5 to 41.8 degrees C for up to 150 min. Pain relief or a sense of well being was observed post-WBH in the first three patients entered in this study. We postulated that WBH might result in increases in the opiate peptide beta-endorphin. Therefore we elected to study prospectively the next six patients entered in this study to test the hypothesis that WBH stimulates the neuroendocrine axis. Results are reported which show thermal induced increases in plasma levels of beta-endorphin as well as prolactin, ACTH and cortisol. PMID- 3036974 TI - Restorative proctocolectomy with ileal reservoir. PMID- 3036976 TI - Multiple adenomatous polyps arising in a continent reservoir ileostomy. AB - In a 29-year-old man who presented with leakage from a continent ileostomy after proctocolectomy, endoscopic evaluation of the reservoir revealed extensive adenomatous polyposis. The polyps were not present at the time of revision of the reservoir 4 years earlier. Because it was not possible to fashion an adequate nipple valve in the presence of so many polyps and the concern over possible malignant transformation, the reservoir was excised. This is the second reported case of polyposis involving a continent reservoir ileostomy. Patients with a continent ileostomy constructed after proctocolectomy for polyposis coli should be evaluated endoscopically at regular intervals. PMID- 3036973 TI - Cachectin/tumor necrosis factor: an endogenous mediator of shock and inflammation. PMID- 3036977 TI - [Tissue response to hydroxylapatite implant in the femurs of rabbits]. PMID- 3036978 TI - Dissociation of human neutrophil activation events by prolonged treatment with amiloride. AB - Human polymorphonuclear leukocytes (PMNs) can be stimulated to release granule contents and to produce superoxide anion. These functional responses are associated with cellular alkalinization and influx of Na+ in exchange for H+. Amiloride is a potassium-sparing diuretic that will inhibit stimulus-induced Na+ H+ exchange and prevent an increase in cell pH. Amiloride has been shown to inhibit a number of protein kinases including the calcium phospholipid-dependent protein kinase. Because PMA, which binds and activates C-kinase, is a potent stimulus of the PMN, this study was undertaken to investigate the effect of prolonged incubation of PMNs with amiloride on PMN stimulation by the chemotactic peptide N-formyl-L-methionyl-L-leucyl-L-phenylalanine (FMLP), the calcium ionophore A23187, opsonized zymosan particles, and the tumor promoter phorbol myristate acetate (PMA). Our results demonstrate that amiloride inhibits superoxide anion production by FMLP, A23187, and opsonized zymosan by causing a slower rate of release and lower maximal release without altering lag time. In contrast, amiloride, despite an inhibition of 22Na+ influx, did not affect superoxide anion production stimulated by PMA. PMN degranulation, phagocytosis, arachidonic acid release, and early influx of calcium were unaffected by preincubation with amiloride. These data suggest that PMN superoxide release induced by FMLP, A23187, and opsonized zymosan is likely modulated by amiloride sensitive Na+-H+ exchange; and phorbol ester-induced superoxide anion release and degranulation by any stimulant do not appear to be modulated by inhibition of an amiloride-sensitive Na+-H+ exchange. PMID- 3036975 TI - Sex and the bowel. AB - Patients with gastrointestinal symptoms may not associate their problems with sexual activities or be unwilling to admit to homosexuality. The diagnosis may be delayed or missed and sexual partners are unlikely to be investigated. To complicate matters further the clinical manifestations of some infections may be markedly altered in a patient whose immune system has been compromised by infection with Human Immunodeficiency Virus. PMID- 3036979 TI - Quantitative measurement of plasma gelsolin and its incorporation into fibrin clots. AB - Gelsolin is a newly recognized actin-binding protein of plasma that severs actin filaments. The concentration of plasma gelsolin was measured by three independent methods: an enzyme-linked immunosorbent assay (ELISA) and two functional assays based on the ability of gelsolin to accelerate the polymerization of actin (a "nucleating" assay) or to sever preformed actin filaments (a "cutting" assay). The gelsolin level in 56 samples of human plasma ranged between 100 and 330 micrograms/ml. The mean plasma concentrations measured by the different assays were ELISA, 207 micrograms/ml; nucleating assay, 233 micrograms/ml; cutting assay, 247 micrograms/ml. The mean serum level of gelsolin was found to be 24% lower than that of plasma when paired samples were examined (183 micrograms/ml). The difference between plasma and serum gelsolin concentrations can be accounted for by a direct interaction between gelsolin and fibrin as shown by the binding of radiolabeled gelsolin to clots made from purified fibrinogen. Further, unlabeled gelsolin inhibits the binding of the labeled species to fibrin, but hemoglobin and albumin do not. Fibrin oligomers, but not fibrinogen, alter the sedimentation characteristics of radiolabeled gelsolin in sucrose gradients. The amount of gelsolin incorporated into the clot is increased if the clots are made in the presence of actin filaments or fibronectin. Thus, serum levels of gelsolin are lower than plasma levels because of an interaction between gelsolin and fibrin clots. PMID- 3036980 TI - A study of three preparations in the treatment of otitis externa. AB - Twenty-seven patients with an initial diagnosis of otitis externa, untreated for the previous 2 weeks, were treated with weekly aural suction toilet and a choice of three topical drops. Two contained combinations of antibiotics and steroids, one of which also contained an anti-fungal agent. The third was boric acid in spirit. The patients were assessed at weekly intervals until a remission was established. From this study it would appear that antibiotic-based drops confer no real advantage over boric acid in spirit. PMID- 3036981 TI - Glomus jugulare tumour masquerading as 'idiopathic' facial nerve palsy. PMID- 3036982 TI - Ductal carcinoma of the parotid gland. AB - A case of ductal carcinoma of the parotid gland is described. The medical literature contains only 13 previous reports on this kind of adenocarcinoma of the parotid gland. The tumour is characterized by its histologic resemblance to ductal carcinomas of the breast and prostate. The course of previously described cases suggests that this tumour has a highly aggressive biological behaviour. PMID- 3036983 TI - Induction of chemotaxis in mouse peritoneal macrophages by activators of protein kinase C. AB - Two activators of calcium and phospholipid dependent protein kinase (protein kinase C), the tumor promoter, 12-O-tetradecanoyl-phorbol-13-acetate (TPA) and the synthetic diacylglycerol, 1-oleoyl-2-acetylglycerol (OAG), were compared as chemotactic agents for mouse peritoneal macrophages. Both of these compounds were found to induce chemotaxis in the macrophages to a similar extent in a time and dose dependent manner. Induction of chemotaxis was observed in the concentration range of 10-100 nM for TPA and 25-250 microM for OAG. Two structurally related synthetic sn 1,2-diacylglycerols, 1,2-dioctanoylglycerol (diC8) and 1,2 didecanoylglycerol (diC10), were also found to be chemotactic for macrophages, while monoacylglycerol (2-monoolein) was inactive. Of the diacylglycerols, OAG was found to be the most active followed by diC8 and diC10. In contrast to TPA, the synthetic diacylglycerols had no effect on superoxide anion release by the cells, suggesting that the mechanism of superoxide anion release by TPA in macrophages is distinct from chemotaxis. Phorbol-12,13-diacetate, a biologically inactive phorbol ester analog that inhibits the binding of TPA to its cellular receptors, inhibited macrophage chemotaxis induced by TPA, the synthetic diacylglycerols, and the complement fragment, C5a. Taken together, our results suggest that chemotaxis in macrophages may be mediated by activation of protein kinase C. PMID- 3036985 TI - Volume dependent polymorphonuclear leukocytes fractions isolated by counterflow centrifugal elutriation: PMN volume does not correlate with PMN age. AB - We have previously reported that human peripheral blood polymorphonuclear leukocytes can be separated into at least six volume dependent fractions using counterflow centrifugal elutriation (CCE). Larger volume was shown to correlate with increased superoxide release with either the chemotactic peptide F-met-leu phe (FMLP) or the tumor promotor phorbol myristate acetate (PMA). To help explain these findings we have examined volume dependent PMN fractions (VDPF) for PMN functions felt to be correlated to PMN age, ie, alkaline phosphatase activity, phagocytosis of opsonized particles, adherence to plastic, and directed movement to FMLP and zymosan activated serum. PMN alkaline phosphatase activity was found to be low in the smallest PMNs, increasing in the PMNs of intermediate size, and then decreasing again in the largest PMN functions. A similar relationship was noted for PMN phagocytic activity. No significant difference among VDPF in adherence to plastic or chemotaxis was found. Furthermore, no difference among VDPF was seen for receptor number or binding affinity of the ligands FMLP or phorbol dibutyrate. Total cellular activity of the NADPH dependent oxidase was, however, 200% greater in the largest compared to the smallest VDPF. Because of the lack of consistent correlation between "age" related PMN function and PMN volume, it is unlikely that PMN volume represents a manifestation of PMN age. It is likely that the increased oxidase activity seen among larger VDPF accounts for the more rapid oxidative burst previously noted. PMID- 3036984 TI - Expression of an activation antigen, Mo3e, associated with the cellular response to migration inhibitory factor by HL-60 promyelocytes undergoing monocyte macrophage differentiation. AB - HL-60 promyelocytic cells acquire the surface expression of the Mo3e antigenic determinant after exposure to PMA or compounds that raise intracellular concentrations of cyclic AMP (dibutyryl cyclic AMP or a combination of cholera toxin and IBMX). The expression of Mo3e by these stimulated HL-60 cells coincides with the development of features of monocyte-macrophage differentiation (characteristic morphology, nonspecific esterase activity, and respiratory burst activity). During in vitro monocyte-macrophage differentiation, HL-60 cells become responsive to migration inhibitory factor (MIF); the MIF responsiveness of differentiated HL-60 cells is blocked by anti-Mo3e monoclonal antibody. These findings further support the relationship between the expression of Mo3e and the cellular response to MIF. PMID- 3036987 TI - Defective production of reactive oxygen intermediates by tumor-associated macrophages exposed to phorbol ester. AB - Macrophages were isolated from poorly immunogenic metastatic sarcomas (mFS6 and MN/MCA1) of C57BL/6 origin. Tumor-associated macrophages (TAM) showed little release of superoxide when exposed to phorbol myristate acetate. When exposed to a phagocytic stimulus (zymosan), TAM released appreciable amounts of superoxide. TAM had a lower number of specific binding sites for phorbol esters than resident or caseinate-elicited peritoneal macrophages, but had normal NADPH-cytochrome C reductase. The tumor environment, possibly through previously demonstrated products of neoplastic cells, may influence the functional status of in situ macrophages and, thus, impair host anti-tumor and anti-microbial defense mechanisms. PMID- 3036986 TI - A high-yield, high-purity elutriation method for preparing human granulocytes demonstrating enhanced experimental lifetimes. AB - A counterflow centrifugal elutriation procedure for preparing human granulocytes is described. This method permits isolation of 1-2 X 10(9) granulocytes with a yield of 70% +/- 7% from a unit of whole blood in 3 h. These cells are 99% +/- 1% pure and 95-99% viable. When compared to cells prepared by a conventional procedure employing hypotonic lysis to remove red cells, the elutriated cells show enhanced oxidative responses resulting from enhanced longevity. Other responses including receptor-mediated uptake of chemoattractant were not significantly different from those of conventionally prepared cells after incubation at 37 degrees C prior to stimulation. The population of elutriated granulocytes, however, was significantly more homogeneous and pure than cells prepared conventionally. PMID- 3036988 TI - Regulation of macrophage-derived fibroblast growth factor release by arachidonate metabolites. AB - The macrophage is a source of many mediators with direct and indirect fibrogenic potential. In this study, release of macrophage-derived fibroblast growth factor (MDGF) activity by murine peritoneal macrophages is examined with regard to its regulation by arachidonate metabolites. Upon stimulation with 10 micrograms/ml lipopolysaccharide (LPS), resident peritoneal macrophages from CBA/J mice released MDGF activity into media rapidly, reaching maximal levels in approximately 1 h. Lysates of these stimulated cells also revealed significantly increased cell-associated MDGF activity, composing 45% of the total assayable activity. This activity, as assayed by radioactive thymidine incorporation by primary cultures of rat lung fibroblasts, was separable from interleukin-1 (IL-1) activity by reverse phase high performance liquid chromatography (HPLC). Furthermore, purified murine IL-1 had no MDGF activity in this assay system. This stimulated MDGF release was enhanced by the cyclooxygenase inhibitors indomethacin, ibuprofen, and aspirin at micromolar concentrations, but inhibited in a dose-dependent manner by prostaglandin E2 (PGE2). On the other hand, nordihydroguaiaretic acid (NDGA), a lipoxygenase inhibitor was inhibitory at 0.1 and 0.4 microM but not at 2.5 microM. Zymosan-stimulated macrophages also markedly increased MDGF release, albeit with a different time course which was characterized by a delay of approximately 7 h before peak levels were attained. Such stimulation, which is known to cause increased lipoxygenase activity, was also inhibited by 0.5 microM NDGA. In contrast, the lipoxygenase pathway products leukotrienes B4 (LTB4) and C4 (LTC4) stimulated MDGF release in a dose-dependent (10(-10)-10(-8) M) manner, with LTC4 being more potent on a per unit dose basis. Stimulation by LTC4 was inhibited by the putative leukotriene receptor antagonist, FPL55712, while LTD4 and LTE4 did not stimulate MDGF release, thus suggesting the mediation of this effect by specific LTC4 receptors. These data suggest also that products of the cyclooxygenase and lipoxygenase pathways are potentially important both as exogenous (ie, derived from cells other than the macrophage itself) and auto- or self-regulators of macrophage MDGF release. This, in turn, implies that cyclooxygenase products are antifibrogenic and important in maintaining or returning to the quiescent or normal state, whereas the lipoxygenase products are profibrogenic and important in induction of fibrosis or wound-healing and tissue repair. Any alteration in the balance between these two pathways may result in either a desirable or a harmful outcome. PMID- 3036989 TI - Similarities in cAMP responses between murine lymphoid cell lines and subsets of mouse lymphocytes. AB - Intracellular cAMP responses to forskolin (which stimulates independently of hormone receptors) and to several hormones were studied in mouse lymphoid cell lines of B- and T-cell origin and in subpopulations of mouse lymphocytes. In the T-cell lines, isoproterenol caused an increase in cAMP amounting at most to 28% of that caused by 100 microM forskolin. In contrast, in the B-cell lines the increase ranged from 50% to over 100%. In 11 of 13 T-cell lines, forskolin at 1 microM concentration potentiated the isoproterenol response five- to tenfold. Forskolin potentiation was also found in thymocytes and peripheral T-cells but not in B-cell lines. The relative increases in intracellular cAMP evoked by isoproterenol, prostaglandin E2 (PGE2) and histamine varied markedly from cell line to cell line. For instance, three lines carrying the T-helper cell marker L3T4 responded only to PGE2 and not to isoproterenol, whereas two cell lines carrying the Ly2 marker gave a higher response to isoproterenol than to PGE2. The results indicate that subsets of lymphocytes respond differently to various hormones and that these differences are maintained in continuous cell lines. PMID- 3036991 TI - Aflatoxin, viral hepatitis and primary liver cancer. AB - Thirty-eight cases of primary liver cancer (PLC) from the Department of Pathology, Charles University, Hradec Kralove were investigated. The viral hepatitis HBsAg antigen was found in the sera of 8 of the 38 patients. Radioimmunoassay demonstrated aflatoxin in 27 of 34 cases. Fluorescence microscopy revealed a fluorescence characteristic of aflatoxin and its fluorescent metabolites in the peritumoral tissue and the tumor for all of the cases studied. This study emphasizes the increasing incidence of PLC over the last 15 years. In addition, this type of cancer now appears to develop in a different manner: reduction in the number of cases with cirrhosis, progressive rise in the number of women affected, and occurrence of PLC in children and adolescents. These observations tend to indicate that aflatoxin might play a significant role in the genesis of PLC. PMID- 3036990 TI - Glucan: mechanisms involved in its "radioprotective" effect. AB - It has generally been accepted that most biologically derived agents that are radioprotective in the hemopoietic-syndrome dose range (eg, endotoxin, Bacillus Calmette Guerin, Corynebacterium parvum, etc) exert their beneficial properties by enhancing hemopoietic recovery and hence, by regenerating the host's ability to resist life-threatening opportunistic infections. However, using glucan as a hemopoietic stimulant/radioprotectant, we have demonstrated that host resistance to opportunistic infection is enhanced in these mice even prior to the detection of significant hemopoietic regeneration. This early enhanced resistance to microbial invasion in glucan-treated irradiated mice could be correlated with enhanced and/or prolonged macrophage (but not granulocyte) function. These results suggest that early after irradiation glucan may mediate its radioprotection by enhancing resistance to microbial invasion via mechanisms not necessarily predicated on hemopoietic recovery. In addition, preliminary evidence suggests that glucan can also function as an effective free-radical scavenger. Because macrophages have been shown to selectively phagocytize and sequester glucan, the possibility that these specific cells may be protected by virtue of glucan's scavenging ability is also suggested. PMID- 3036992 TI - Five-year survival following chemotherapy of non-small cell lung cancer. AB - Five-year survival of patients with inoperable non-small cell lung carcinoma is rare. We report a patient with inoperable large cell lung carcinoma who is apparently cured after receiving a course of combination chemotherapy. As the primary treatment of inoperable non-small cell carcinoma, we recommend combination chemotherapy over radiation therapy because: Chemotherapy gives a response rate of 30-40%; Quality of survival following chemotherapy appears better than following radiation therapy; Chemotherapy following radiation therapy appears to be ineffective. PMID- 3036993 TI - Time-dependent effect of intracranial injection of cyclic AMP on blood glucose homeostasis. PMID- 3036994 TI - Single germline VH and V kappa genes encode predominating antibody variable regions elicited in strain A mice by immunization with p-azophenylarsonate. AB - We have cloned and sequenced the predominant germline V kappa gene segment expressed by B cells of strain A origin that synthesize antibodies with specificity for Ars. In hybridomas synthesizing anti-Ars antibodies, this V kappa gene segment (V kappa IdCR) has been found exclusively associated with the J kappa 1 gene segment without exhibiting junctional sequence variation. Sequence comparisons of the germline V kappa IdCR gene with expressed derivatives reveals that the latter frequently contain somatically introduced amino acid replacements. Taken together with results of previous structural analyses, these results show that the predominant population of IdCR+ V regions elicited in the secondary immune response is encoded by one or two combinations of V gene segments, has little junctional diversity, and is extensively diversified by somatic mutation in both heavy and light chains. PMID- 3036995 TI - Inducibility of Ia antigen on astrocytes by murine coronavirus JHM is rat strain dependent. AB - Inducibility of Ia molecules on cultivated astrocytes by JHM virus correlates with demyelinating disease susceptibility of animals from which these astrocytes are derived. On the contrary, class I induction of both astrocytes and oligodendrocytes occurs as a consequence of normal cultivation procedures in both susceptible and resistant strains. Increased expression of class I antigens on rat astrocytes and oligodendrocytes is not related to JHM viral infection as it is in the mouse. These data indicate that strain differences in Ia inducibility, rather than inducibility of class I antigens, by JHM virus may explain higher levels of T cell-mediated damage to myelin during infection in susceptible rat strains compared with resistant strains. PMID- 3036996 TI - Calcium and inositolphosphates in the activation of T cell-mediated cytotoxicity. AB - Reports from a number of laboratories have shown that mAbs against the T3-Ti receptor complex cause an increase in cytosolic-free Ca2+ [( Ca2+]i) and the hydrolysis of phosphatidylinositolbisphosphate (PIP2) in CTLs. In the present report we show that activation of CTLs by their specific targets causes: (a) release of Ca2+ from intracellular stores; (b) transient formation of inositol trisphosphate (InsP3); and (c) an increased permeability to Ca2+ of CTL plasma membrane. Killing of unrelated targets could be induced by cocentrifugation of the unrelated targets with CTLs in the presence of A23187 or PMA. We conclude that: (a) activation of CTLs by specific antigens triggers the generation of the same intracellular mediators generated by stimulation of lymphocytes with anti-T3 Ti receptor antibodies and/or with polyclonal mitogens; and (b) intracellular signals that mediate the delivery of the lethal hit by CTLs are indistinguishable from those that induce cell proliferation. PMID- 3036997 TI - A low polymorphic mouse H-2 class I gene from the Tla complex is expressed in a broad variety of cell types. AB - We have previously described the isolation of pH-2d-37, a cDNA clone that encodes a so far unknown, poorly polymorphic, class I surface molecule. We report here the isolation of the corresponding gene, its nucleotide sequence, and its localization in the Tla region of the murine MHC. Using a RNase mapping assay, we have confirmed that the second domain coding region of the 37 gene displays very limited polymorphism, and that the gene is transcribed in a broad variety of cell types, in contrast to the genes encoding the known Qa and TL antigens. Possible functions are discussed. PMID- 3036999 TI - Role of vitamin D in the development of the chick embryo. AB - Chick embryos of various ages and 1-day-old chicks were injected with different doses of 1,25(OH)2D3 and the concentrations of Ca and Pi in their blood were determined 24 hr after. It was confirmed that the dose required to induce hypercalcemia and hypophosphatemia was much lower in the case of 15-day-old embryos than in younger ones. The age of maximal response corresponds to the period when the chorionic epithelium is fully differentiated. Newly hatched chicks did not respond even with doses 20 times higher than those used in 15-day old embryos. These findings support the idea that the humoral changes produced by 1,25(OH)2D3 result from increased resorption of calcium from the shell and that the chorionic epithelium is instrumental in this phenomenon. The autoradiographic study of tissues from embryos injected with tritiated 1,25(OH)2D3 showed that the exposure time required to detect nuclear concentration of radioactivity was much shorter in the case of "villus-cavity" cells than in any other tissue (kidney, intestine, bone, parathyroid glands) in which target cells were recognized. The above results are interpreted as indicating that the chorionic epithelium is the main target for 1,25(OH)2D3. The histological study of various organs from embryos injected with large doses of 1,25(OH)2D3 showed generalized infiltration of spleen, liver, and bursa of Fabricius with eosinophilic granulocytes. It has been suggested that 1,25(OH)2D3 influences the proliferation and differentiation of leukocytes; the chick embryo might be a good model for further exploration of this problem. PMID- 3037000 TI - Activity of putative pulmonary ornithokallikrein and angiotensin-converting enzyme during the onset of lung respiration in the chick embryo. AB - We have assayed a proteinase from chicken lungs, the properties of which were comparable to those of ornithokallikrein, the enzyme that catalyzes the formation of ornithokinin. The activity of this pulmonary proteinase showed an accelerated increase during the development of lung respiration (paranatal period). In contrast, the increase in the activity of angiotensin-converting enzyme (ACE), which degrades kinin, was not enhanced at this developmental stage. L-thyroxine administration accelerated the onset of lung respiration and the associated changes in the activities of ornithokallikrein and ACE. Thiourea, on the other hand, retarded the onset of lung respiration. Concomitantly, there was no pronounced increase in ornithokallikrein activity and no attenuation of the increase in ACE activity. Furthermore, the development of lung circulation, as measured by pulmonary hemoglobin concentration, paralleled the ornithokallikrein activity. The results suggest that the increase in ornithokallikrein activity and the suppression of ACE activation during the paranatal period help adapt the embryo to the neonatal period. PMID- 3037001 TI - Gene transfer in swine embryos by injection of cells infected with retrovirus vectors. AB - Key components for gene transfer to swine embryos using an avian retrovirus are described. A replication-defective reticuloendotheliosis (REV) viral vector can infect and be expressed in pig embryo fibroblasts (PEF). Infection with a replication-competent vector (REV-A) indicates a presumptive block to viral replication in PEF. Swine embryos obtained at the morula stage can be cultured in vitro to the blastocyst stage, injected with retrovirus helper cells or quail embryo fibroblasts producing REV, and transferred to recipient swine with survival to at least 6 wk of gestation. PMID- 3036998 TI - Four Ia invariant chain forms derive from a single gene by alternate splicing and alternate initiation of transcription/translation. AB - We determined the structural basis for the presence of electrophoretically distinct, antigenically-related forms of invariant chains in Ia oligomers, and established the mechanisms by which they can be expressed from a single gene. S1 nuclease protection assays indicated that, in B cells, transcription of this gene initiates at a minimum of three sites. Thus, unlike previously thought, invariant chain mRNAs have heterogeneous 5' untranslated segments that may differentially affect initiation of translation. Further, restriction mapping and nucleotide sequencing of cDNAs revealed two kinds of invariant chain mRNAs differing by an internal coding segment of 192 bp. This segment represents an alternatively spliced exon, as demonstrated by nucleotide sequencing of corresponding genomic regions. The exon (exon X) encodes a cysteine-rich stretch of 64 amino acids near the COOH terminus that displays a striking and surprising homology to an internal amino acid repeat of thyroglobulin, suggesting an evolutionary mechanism of exon shuffling. Transient expression of cDNAs indicated that both types of alternatively spliced mRNAs contain two in-frame AUGs functioning as alternate start sites for translation. Thus, transfections with exon X-lacking cDNAs resulted in the expression of Mr 33,000 and 31,000 proteins, detected by immunoprecipitation with anti-invariant chain antisera, and identical by two dimensional gel (2-D) analyses to the B cell invariant-chain forms gamma 1 (Mr 31,000), gamma 2, and gamma 3 (Mr 33,000). Similarly, exon X-containing cDNAs expressed Mr 43,000 and 41,000 proteins, also identical by 2-D migration to Ia associated proteins. Thus, human Ia molecules contain four forms of invariant chain of closely related but nonidentical primary structure that are generated from a single gene by a complex pattern of alternate transcriptional start, exon splicing, and translational start. PMID- 3037002 TI - Comparison of rotazyme, Slidex Rota-Kit, counterimmunoelectrophoresis with solid phase immune electron microscopy for the detection of human rotavirus. PMID- 3037004 TI - Changes of serum levels of lipid components and apo-lipoproteins after hepatectomy in patients with hepatocellular carcinoma. PMID- 3037003 TI - Antibacterial activity of ampicillin alone and in combination with sulbactam correlated with beta-lactamase production. PMID- 3037006 TI - Identification of human rotaviruses in stools from children with gastroenteritis. PMID- 3037005 TI - Mode of synchronized DNA synthesis in the nuclei of Sendai virus-induced multinuclear cells. PMID- 3037007 TI - Diffusional solute flux during osmotic water flow across the human red cell membrane. AB - The effect of solvent drag on the unidirectional efflux of labeled water, urea, and chloride from human red cells was studied by means of the continuous flow tube method under conditions of osmotic equilibrium and net volume flow. Solvent (water) flow out of cells was created by mixing cells equilibrated in 100 mM salt solution with a 200-mM or 250-mM salt solution, while flow of water into cells was obtained by equilibrating the cells in the higher concentration and mixing them with the 100-mM solution. Control experiments constitute measurements of efflux of [14C]ethanol in normal cells and 3H2O in cells treated with p chloromercuribenzosulfonate under the conditions described above. In both instances, the solute is known to penetrate the membrane through nonporous pathways. As anticipated, the tracer flux of neither urea nor chloride showed any dependence on net solvent flow, regardless of the direction. If one assumes the recently reported reflection coefficient for urea of 0.7, the urea tracer flux should change by at least 24% under volume flow conditions. Since such changes would be easily detected with our method, we conclude that the pathways for water, for urea, and for chloride are functionally separated. PMID- 3037009 TI - Analysis of the bond between encephalomyocarditis virus and its human erythrocyte receptor by affinity chromatography on virus-sepharose columns. AB - Affinity chromatography was used to analyse the bond between encephalomyocarditis (EMC) virus and glycophorin, the receptor for EMC virus on human erythrocytes. Between 60 and 80% of glycophorin added to virus-Sepharose columns was retained compared with 10 to 20% retention on glycine-Sepharose columns. Elution with 0.2 M-NaCl released about 80 to 90% of the retained glycophorin from virus-Sepharose columns but little from glycine-Sepharose. Glycophorin remaining on either the virus or glycine columns after 0.2 M-NaCl treatment was released with Triton X 100, suggesting that this material was aggregated and trapped non-specifically. The sensitivity of the EMC virus-glycophorin bond to 0.2 M-NaCl was confirmed by showing that the radiolabelled EMC virus that bound to human erythrocyte membranes in 0.02 M-phosphate buffer was released when washed with this buffer containing 0.2 M-NaCl. It was concluded that weak ionic interactions were involved in this virus-receptor bond. Treatment of glycophorin with neuraminidase prevented it binding to EMC virus-Sepharose indicating the requirement for sialic acid for receptor activity. However, other sialylated molecules did not bind. Only one chymotryptic peptide of glycophorin (CH0) bound to EMC virus-Sepharose, confirming that this peptide contains the virus-binding site as had been previously suggested using other techniques. The effects of two non-ionic detergents and two anionic detergents on the virus-glycophorin bond were examined. In each case a very low concentration of detergent disrupted the bond and the concentration required was in parallel with that which dissolved erythrocyte membranes. It appears that multivalent glycophorin aggregates are required for stable bond formation. PMID- 3037008 TI - The complete nucleotide sequence of coxsackievirus B4 and its comparison to other members of the Picornaviridae. AB - The genome of the prototype stain of coxsackievirus B4 (J.V.B. Benschoten) has been cloned in Escherichia coli and its complete nucleotide sequence determined. Excluding the poly(A) tract, the RNA genome is 7395 nucleotides in length and appears to encode a single polyprotein of 2183 amino acids. The predicted amino acid sequence of the polyprotein shows close homology (88%) to that of the previously sequenced coxsackievirus B3 and to certain regions of the polyproteins of the polioviruses and human rhinovirus 14. This allows identification of putative polyprotein cleavage signals, antigenic domains and other structural features likely to be important to the biological integrity of the virus. PMID- 3037010 TI - Conservation in vivo of protease cleavage sites in antigenic sites of poliovirus. AB - Polioviruses possess three major antigenic sites which have been located chemically and structurally on the particle. One of these sites, designated site 1, is strongly immunodominant for serotype 3, but highly immunorecessive for type 1. We report that monoclonal antibodies directed against site 1 of type 1 poliovirus may be isolated by an altered route of immunization of the donor mice. Site 1 is shown to be highly variable for type 1, but highly conserved for type 3 poliovirus, although the converse would be predicted from their immunodominance. The evidence presented suggests that the antigenic conservation is associated with a strong selective pressure for a proteolytic cleavage site within site 1 of type 3. As proteolytic cleavage results in the loss of the antigenicity of site 1 the presence of the cleavage site in a virus replicating in the gut in the presence of proteases would protect the virus from neutralizing antibodies directed against uncleaved site 1. The conservation of the site in type 3 is thus consistent with the view that site 1 is a significant target of a human as well as a murine immune response against type 3. PMID- 3037011 TI - The predicted primary structure of the peplomer protein E2 of the porcine coronavirus transmissible gastroenteritis virus. AB - The complete nucleotide sequence of cloned cDNAs containing the E2 glycoprotein encoding region of the genome of transmissible gastroenteritis virus (TGEV) has been determined. A single large translatable frame of 4.3 kb starting at 8.2 kb from the 3' end of the genome was identified. Its deduced amino acid sequence contains the characteristic features of a coronavirus peplomer protein: the precursor polypeptide of TGEV E2 is 1447 residues long (i.e. 285 longer than the avian infectious bronchitis coronavirus spike protein); partial N-terminal sequencing demonstrated that a putative secretory signal sequence of 16 amino acids is absent in the virion-associated protein; the predicted mol. wt. of the apoprotein is 158K; most of the 32 potential N-glycosylation sites available in the sequence are presumed to be functional to account for the difference between this and the experimentally determined value (200K to 220K); a typical hydrophobic sequence near the C terminus is likely to be responsible for anchoring the peplomer to the virion envelope. PMID- 3037013 TI - Structural and immunological characterization of the intracellular forms of an abundant 68,000 Mr human cytomegalovirus protein. AB - Murine monoclonal antibodies and polyvalent monospecific antisera reactive with an abundant 68,000 Mr (p68) human cytomegalovirus (CMV) virion protein were used to characterize this protein within CMV-infected cells. The protein was found to partition within the nucleus and cytoplasm of infected cells. Pulse-chase analysis indicated the p68 was degraded into three proteins of 52,000, 51,000 and 50,000 Mr which were found only within infected cells. Both cellular forms as well as the virion p68 were phosphorylated but non-glycosylated. The p68 was synthesized shortly after infection and in the presence of cytosine arabinoside, an inhibitor of viral DNA replication. Studies with monospecific antisera and a panel of monoclonal antibodies specific for the p68 suggested that this protein was not expressed on the surface of infectious virions or infected cells. PMID- 3037012 TI - Purification of papillomavirus structural polypeptides from papillomas by immunoaffinity chromatography. AB - A broadly cross-reactive monoclonal antibody directed against papillomavirus, coupled to immunoaffinity columns, was used to isolate bovine papillomavirus type 1 (BPV-1) and human papillomavirus type 1 (HPV-1) structural polypeptides from homogenates of productively infected cells. One of the polypeptides isolated from bovine fibropapillomas appeared to be the BPV-1 major capsid protein since it had a mol. wt. of 54K and was reactive by Western blots with papillomavirus genus- and BPV-1 type-specific rabbit antibodies as well as monoclonal antibodies cross reactive with BPV-1/BPV-2 and BPV-1/deer PV. A polypeptide from human plantar warts similarly appeared to be the major capsid component since it also had a mass of 54K to 55K and reacted with papillomavirus genus- and HPV type-specific rabbit antibodies. By using this technique structural viral polypeptides from papillomavirus-induced lesions containing readily detectable viral structural antigens but relatively few virus particles, such as seen with mucosotropic HPVs can now be isolated for mapping of virus-specific epitopes. PMID- 3037014 TI - Replacement of glycoprotein B gene sequences in herpes simplex virus type 1 strain ANG by corresponding sequences of the strain KOS causes changes of plaque morphology and neuropathogenicity. AB - DNA sequences encoding glycoprotein B (gB) derived from herpes simplex virus type 1 (HSV-1) strain KOS321 were transferred to HSV-1 ANG. In cotransfection experiments the cloned HSV-1 KOS BamHI G fragment served as donor, and genomic DNA of two ANG variants as recipients. One of these variants, HSV-1 ANG path, expresses gC and the other, C18, was a spontaneous gC-negative mutant. Both ANG strains are of the syncytial (syn) phenotype whereas HSV-1 KOS321 is non syncytial (syn+). Recombinants were identified by means of a monoclonal antibody which selectively recognizes gBKOS. Among the HSV-1 ANG path/gBKOS recombinants, the majority displayed an altered plaque morphology, i.e. they were of the syn+ phenotype. In contrast all of the C18/gBKOS recombinants were of the syn phenotype. The possibility that the mutant C18 carries a syn mutation not present in the parental strain could be excluded. Marker transfer experiments involving subfragments of the gB gene mapped the syn mutation of HSV-1 ANG path to a locus within the gene that has been previously termed syn 3. Subclones of HSV-1 ANG path were established either directly or after intermittent transfection or cotransfection with the KOS BamHI G fragment. The pathogenicity in mice of these clones was compared. The data obtained indicated that at high frequency, the BamHI G fragment confers apathogenicity. PMID- 3037016 TI - Herpes simplex virus type 2 glycoprotein biogenesis: effect of monensin on glycoprotein maturation, intracellular transport and virus infectivity. AB - The ionophore monensin inhibited the formation of herpes simplex virus type 2 (HSV-2) particles by about 30% but the yields of infectious particles were reduced to 5% and 1% for cell-associated and extracellular virus, respectively. The presence of monensin did not affect the processing of the two viral glycoproteins gB-2 and gG-2. However, two other glycoproteins, gC-2 and gD-2, were not processed to their fully mature forms in the monensin-treated cells and only the faster moving pgC-2 and pgD-2 were detected. The cell-associated virus particles contained the glycoproteins gB-2, gC-2, gD-2 and gG-2, whereas the extracellular virus particles contained only gG-2 glycoprotein. These results suggest that HSV-2 particles containing all the viral glycoproteins are transported via the Golgi apparatus to the cell surface but that virus particles containing only gG-2 may follow a different pathway for transport and release. PMID- 3037015 TI - The products of gene US11 of herpes simplex virus type 1 are DNA-binding and localize to the nucleoli of infected cells. AB - We have used antisera raised against synthetic oligopeptides to characterize the protein products from herpes simplex virus type 1 gene US11. These antisera recognized predominantly polypeptides of apparent molecular weight 21,000 and 22,000, but also polypeptides of apparent molecular weight 17,500, 15,000, 14,000 and 11,000. Tryptic peptide fingerprint analysis confirmed that these polypeptides were all closely related. The 21,000 and 22,000 molecular weight polypeptides were shown to be DNA-binding proteins, and immune electron microscopy demonstrated their strong localization within nucleoli of infected cells. PMID- 3037017 TI - Virus-specific IgM and IgG antibody production by B cells during herpes simplex virus type 2-induced immunosuppression as analysed by an immunospot assay. AB - The mechanism of herpes simplex virus (HSV)-2-induced immunosuppression was analysed by determination of the number of IgM and IgG antibody-secreting B cells in female BALB/c mice using an immunospot assay. Primary HSV-1 or -2 as well as homologous or heterologous booster infections at different times were performed. In accordance with earlier results on humoral antibody generation, in contrast to HSV-1, HSV-2 induced only very low numbers of antibody-producing B cells in dose response experiments. They appeared late after infection compared to HSV-1. Despite a homologous humoral booster reaction against HSV-1 at day 8 no IgM- or IgG-secreting cells in the spleen could be detected. This non-reactivity of the spleen had vanished 10 days later, when secondary reactions of B cells could be observed. Secondary infections with a high homologous dose of HSV-2 after a low primary dose produced only a low booster response of IgG-secreting B cells. Suppression of humoral antibody production induced by HSV-2 (high dose) waned after more than 50 days, indicating that the HSV-2-induced suppression did not impair antigen presentation or memory cell generation. PMID- 3037019 TI - Immune response to rotavirus polypeptides after vaccination with heterologous rotavirus vaccines (RIT 4237, RRV-1). AB - The antibody response to individual rotavirus polypeptides after vaccination with live attenuated heterologous rotavirus vaccines was studied with a radioimmunoprecipitation assay (RIPA). Following vaccination with bovine rotavirus strain RIT 4237 and rhesus monkey rotavirus strain RRV-1, an antibody response was demonstrated mainly against the inner capsid polypeptides VP2 and VP6 and to a lesser extent against VP3/VP4. There was no qualitative difference between the vaccines in the antibody response; both vaccines induced antibodies against the same polypeptides. Quantitatively the antibody response to RRV-1 was somewhat stronger. All prevaccination sera contained by RIPA low levels of rotavirus antibodies. The effect of these antibodies on the 'take' of the vaccines is discussed. PMID- 3037020 TI - Recovery of herpes simplex virus from the corneas of experimentally infected rabbits. AB - Rabbits were inoculated in the left cornea with one of three strains of herpes simplex virus (HSV) i.e. HSV-1 strain 17, HSV-1 strain McKrae, HSV-2 strain HG52, or with and HSV-1 McKrae/HSV-2 HG52 recombinant R40/2. Fifty-nine to 67 days after inoculation trigeminal ganglia and corneas were explanted and screened for release of infectious virus. Virus was isolated from all left trigeminal ganglia after organ culture irrespective of viral strain. Virus was isolated from three of 16 corneas of animals inoculated with HSV-1 strain McKrae and from one of four corneas from animals inoculated with the HSV-1/HSV-2 recombinant. The isolation of HSV from explanted corneas, after between 15 and 35 days in organ culture suggests that the cornea may be a site additional to dorsal root ganglia where latent HSV can reside in rabbits. PMID- 3037018 TI - Demonstration of bovine viral diarrhoea virus in peripheral blood mononuclear cells of persistently infected, clinically normal cattle. AB - Peripheral blood mononuclear cells (PBL) from cattle known to be persistently viraemic with bovine viral diarrhoea virus (BVDV) following a foetal infection, were examined for the presence of viral antigens and cell-associated infectious virus. Using immunocytochemical techniques, physical separations of PBL subsets and virus isolation techniques (directly and by cocultivation) it was found that infection occurred in B and T lymphocytes, monocytes, and a group of cells designated null cells for lack of more specific classification. The latter three groups also supported viral replication, as infectious virus could be isolated from enriched cell populations. BVDV-like particles in cytoplasmic vesicles of PBL subsets were detected by electron microscopy. PMID- 3037021 TI - The genome of caprine herpesvirus 1: genome structure and relatedness to bovine herpesvirus 1. AB - Caprine herpesvirus 1 (CapHV-1) DNA was examined by electron microscopy, restriction site mapping and homology studies with bovine herpesvirus 1 (BHV-1) DNA. Although the restriction site maps differed significantly, we showed that the genome structures of CapHV-1 and BHV-1 were identical and that the DNAs shared a high degree of base sequence homology. PMID- 3037023 TI - EPR studies of copper(II) and cobalt(II) complexes of adriamycin. AB - Cu2+ and Co2+ complexes of adriamycin (ADM) in aqueous solutions have been examined using EPR spectroscopy. An appreciable amount of Cu2+ and Co2+ complexes formed in the solutions were found to be in the EPR silent associated form, where the metal ions are antiferromagnetically coupled. The associated form of the Cu2+ complex may be neither a simple dimer nor coordination polymer but aggregates of a stacked type. Formation of a complex having Cu2+-ADM stoichiometry of 1:2 was observed for the solutions containing excess of ADM as an EPR observable species. The complex having Cu2+-ADM stoichiometry of 1:1 was not observed directly by EPR, but the presence of the complex is undeniable, especially at low pH range so far as large excessive ADM is not present. The Co2+ complex of ADM observed by EPR is in the high-spin (S = 3/2) state and may have a coordination structure of tetragonal symmetry. The EPR spectra of these complexes apparently show that the Cu2+ and Co2+ ions are bound at the carbonyl and phenolate oxygen in the 1,4 dihydroxyanthraquinone moiety and the amino nitrogen in the sugar part does not seem to participate in the coordination to the metal ions. PMID- 3037022 TI - The effect of pH on the kinetics of iron release from diferric ovotransferrin induced by pyrophosphate. AB - Pyrophosphate-induced iron release from diferric ovotransferrin follows biphasic kinetics in the pH range from 6.6 to 8.6 except at pH 8.0 where the kinetics become monophasic. The rates of formation of the four molecular species, Fe2OT, FeOTN, FeOTC, and ApoOT, were studied by urea gel electrophoresis and the four microscopic rate constants were calculated at various pH values. Below pH 8.0, these intrinsic rate constants for iron release from Fe2OT follow the order k2N greater than k1N greater than k2C greater than k1C. Each constant diminishes almost proportionally with an increase in pH with the faster rate constants being affected more by the fall in hydrogen ions than the slower ones. Around pH 8.0 the four rates are approximately equal, resulting in monophasic kinetics. However, the rate constants from the C-site become faster than that from the N site at pH above 8.2. At low pH, there is a marked preference for iron to be released from the N-site rather than from the C-site and such preference becomes less distinct as pH increases. A rather weak positive cooperativity between the two sites is demonstrated in pH between 6.8 and 7.8. The ligand responsible for the transition from biphasic to monophasic kinetics at pH 8.0 is not known. It is possible that there are different anions such as [CO3(2-)] and [HCO3-] at the two iron-binding sites, which might explain the preferential rates of iron release from these sites during protonation. PMID- 3037024 TI - Blockade of receptor-mediated cyclic GMP formation by hydroxyeicosatetraenoic acid. AB - Receptor-mediated cyclic GMP formation in N1E-115 murine neuroblastoma cells appears to involve oxidative metabolism of arachidonic acid. Evidence in support of this includes the blockade of this response by lipoxygenase inhibitors, e.g., eicosatetraynoic acid (ETYA) or other metabolic perturbants, e.g., methylene blue. It was recently discovered that the lipoxygenase products 15 hydroxyeicosatetraenoic (15-HETE) acid and 12-HETE, like ETYA, were inhibitors of M1 muscarinic receptor-mediated cyclic GMP formation. In the present report, the effects of monoHETEs are explored in more detail, particularly with regard to the function of the muscarinic receptor. Like 12-HETE and 15-HETE (IC50 = 13 and 11 microM, respectively), 5-HETE inhibited the cyclic GMP response to the muscarinic receptor (IC50 = 10 microM). All three of these monoHETEs were shown also to be inhibitors of the cyclic GMP responses to receptors stimulated by carbachol, histamine, thrombin, neurotensin, and bradykinin. 15-HETE was shown to inhibit the muscarinic receptor-mediated response in a complex manner (apparent noncompetitive and uncompetitive components; IC50 = 18 and 2 microM, respectively). 15-HETE did not inhibit either the M1 muscarinic receptor stimulated release of [3H]inositol phosphates from cellular phospholipids or the M2 muscarinic receptor-mediated inhibition of hormone (prostaglandin E1)-induced AMP formation. It seemed possible that the monoHETEs could enter into biochemical pathways for arachidonate in N1E-115 cells. [3H]Arachidonate and the three [3H] monoHETEs all rapidly labeled the membrane lipids of intact N1E-115 cells, with each [3H]eicosanoid producing a unique labeling profile. [3H]15-HETE labeling was noteworthy in that 85% of the label found in the phospholipids was in phosphatidylinositol (PI;t1/2 to steady state = 3 min). Exogenous 15-HETE inhibited the labeling of PI by [3H]arachidonate (IC50 = 28 microM) and elevated unesterified [3H]arachidonate levels. Thus, the mechanism of blockade of receptor mediated cyclic GMP responses by monoHETEs is likely to be more complex than the simple inhibition of cytosolic mechanisms, e.g., generation of a putative second messenger by lipoxygenase, and may involve also alterations of membrane function accompanying the redistributions of esterified arachidonate. PMID- 3037025 TI - Expression of glial fibrillary acidic protein in rat C6 glioma relates to vimentin and is independent of cell-cell contact. AB - Glial fibrillary acidic protein (GFAP) was induced in rat C6 glioma cells grown in M199 and HAM F10 media by addition of 1 mM dibutyryl cyclic AMP. The amount of GFAP per cell increased 7- and 33-fold in M199 and HAM F10 media, respectively. GFAP could be induced in each phase of the cell culture except for the lag phase, where GFAP synthesis was delayed until the onset of the logarithmic growth. The induction took place under conditions where the total protein content of the cell decreased. Measurement of the amount of vimentin indicated that GFAP was induced under conditions of low vimentin concentration. Our results do not support the hypothesis that GFAP induction depends on cell-cell contact or cell proliferation. They indicate a shift from vimentin to GFAP synthesis by an as yet unknown mechanism. PMID- 3037026 TI - Agonist-induced regulation of adrenoceptor subtypes in cerebral cortical slices. AB - Cerebral cortical slices from rat brain were incubated at 37 degrees C for 2 h in the presence of isoproterenol, noradrenaline, or adrenaline, and binding affinities and densities of adrenoceptor subtypes were subsequently examined in homogenized tissue. The density of alpha 2- and total beta-adrenoceptors was estimated using the radioligands [3H]rauwolscine and [3H]dihydroalprenolol (DHA), respectively. The percentages of beta 1- and beta 2-adrenoceptors were defined by inhibiting the binding of [3H]DHA with the beta 1-selective antagonist metoprolol. Exposure of slices to noradrenaline and adrenaline significantly decreased the maximal number of binding sites (Bmax) of alpha 2-adrenoceptors (48 and 37% respectively) without significantly affecting affinity; isoproterenol had no effect. Exposure to isoproterenol, noradrenaline, and adrenaline significantly decreased the Bmax of beta-adrenoceptors (by 60, 34, and 24%, respectively) but did not affect the affinity. Isoproterenol and adrenaline significantly decreased the density of beta 1-adrenoceptors by 75 and 24% and beta 2-adrenoceptors by 23 and 28%, respectively. Noradrenaline significantly decreased the density of beta 1-adrenoceptors by 42% without affecting the number of beta 2-adrenoceptors. These findings indicate that subtypes of adrenoceptors in rat cerebral cortex are differentially regulated by adrenergic agonists. PMID- 3037028 TI - Characterization of kappa opioid receptors in neurosecretosomes from bovine posterior pituitary. AB - The binding properties of opioid receptors on isolated nerve terminals (neurosecretosomes) from bovine posterior pituitaries were characterized. Both [3H]etorphine and [3H]ethylketocyclazocine ([3H]EKC) showed high-affinity binding with complex binding isotherms, consistent with the presence of multiple classes of binding sites. [D-Ala2,D-Leu5]enkephalin showed no specific binding and failed to displace [3H]etorphine at high concentrations, indicating the absence of mu, delta, or benzomorphan (kappa 2) sites. Mathematical modelling of the data suggested the presence of three classes of binding sites. The first was of high affinity with Kd values of 0.9 and 2.0 nM for etorphine and EKC, respectively. The second class of sites appeared to bind etorphine with a KD of 150 nM, and EKC with extremely low affinity (unmeasurable binding). The third class of sites was characterized by KD values of 7 and 2 microM for etorphine and EKC, respectively. These results indicate that the nerve terminals of bovine posterior pituitary contain opioid binding sites of the kappa type. Furthermore, these binding sites appear heterogeneous, consisting of at least two and possibly more subtypes or states. PMID- 3037027 TI - Triethyllead inhibits gamma-aminobutyric acid binding to uptake sites in synaptosomal membranes. AB - Triethyllead (TEL), the active metabolite of tetraethyllead, was shown previously to inhibit selectively high-affinity Na+-dependent uptake of gamma-aminobutyric acid (GABA) into cerebrocortical synaptosomes. Such inhibition was not related to the Na+ gradient, Na+,K+-ATPase activity, [Cl-], or energy charge. We report here that TEL inhibits GABA binding to the presynaptic transporter involved in Na+ dependent uptake. Scatchard plot analysis of Na+-dependent [3H]GABA binding to a highly purified synaptic plasma membrane preparation revealed that 25 microM TEL reduced the Bmax by 44%, leaving the KD unchanged. This binding was reversible and predominantly involved membrane uptake sites, as characterized by pharmacological specificity to GABA ligands. Approximately 85% of specific GABA binding was considered membrane uptake site binding, as indicated by sensitivity to nipecotic acid and diaminobutyric acid, with relative insensitivity to muscimol, bicuculline methiodide, baclofen, and beta-alanine. With respect to previous data, these finding suggest that TEL inhibits Na+-sensitive high affinity GABA uptake by interfering with GABA binding to its presynaptic transporter. PMID- 3037029 TI - Nerve growth factor receptors in human neuroblastoma cells. AB - Receptors for the nerve growth factor protein (NGFR) present in the human neuroblastoma cell line LAN-1 were characterized. LAN-1 cells display high affinity (type I, with KD value of 5.9 X 10(-11) M) and low-affinity (type II, with KD value of 9.2 X 10(-9) M) binding to NGF. NGFR were fractionated by preparative isoelectric focusing in a granulated gel (PEGG). High-affinity binding was found in the 5.9-6.2 pH region of the PEGG, and low-affinity binding in the 4.6-4.8 and 8.8-9.3 pH ranges. After further analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) we observed both 92.5- and 200-kDa molecular species associated with NGF binding activity. The 200-kDa protein was found in fractions displaying high-affinity NGF binding and the 92.5 kDa protein in fractions displaying low-affinity NGF binding. Equilibrium binding analysis of NGF in PEGG fractions confirmed the presence of two specific saturable binding sites with KD values similar to those observed for whole dissociated cells. When NGFR II activity from the acidic region of the PEGG chromatogram was incubated with NGFR II from the basic region of the PEGG chromatogram, there was no change in NGF binding or in the number of apparent NGF receptors. However, incubation of these same fractions with a fraction having only NGFR I showed an apparent increase in high-affinity NGF binding and a decrease in low-affinity NGF binding. Immunoprecipitation of this "mixed" fraction and analysis on SDS-PAGE under reduced and nonreduced conditions showed 200-kDa and 92.5-kDa proteins under nonreduced conditions and a 92.5-kDa protein under reduced conditions. Our findings are consistent with the hypothesis that there are two distinct NGF receptors in NGF-responsive cells. The interconvertibility of low- and high-affinity receptors and the possible existence of a modulator type protein or of "silent" type receptors are also in agreement with our findings. PMID- 3037030 TI - Thiamine triphosphate and membrane-associated thiamine phosphatases in the electric organ of Electrophorus electricus. AB - The main electric organ of Electrophorus electricus is particularly rich in thiamine triphosphate, which represents 87% of the total thiamine content in this tissue. The thiamine pyrophosphate concentration, however, is very low in the eel electric organ and skeletal muscle as compared with other eel or rat tissues. Furthermore, electroplax membranes contain a whole set of enzymes responsible for the dephosphorylation of thiamine tri-, pyro- and monophosphate. Thiamine triphosphatase has a pH optimum of 6.8 and is dependent on Mg2+. The real substrate of the enzyme is probably a 1:1 complex of Mg2+ and thiamine triphosphate. Thiamine pyrophosphatase is activated by Ca2+. The apparent Km for thiamine triphosphate and Vmax are found to be, respectively, 1.76 mM and 5.95 nmol/mg of protein/min. Thiamine triphosphatase activity is inhibited at physiological K+ concentrations (up to 90 mM) and increasing Na+ concentrations (50% inhibition at 300 mM). ZnCl2 (10 mM) inhibits 90% of the enzyme activity. ATP and ITP are also strongly inhibitory. No significant effect of neurotoxins is seen. Membrane-associated thiamine triphosphatase is affected differently by proteolytic enzymes and is partially inactivated by pretreatment with phospholipase C and neuraminidase. The physiological significance of thiamine triphosphatase is discussed in relation to a specific role of thiamine in the nervous system. PMID- 3037031 TI - Angiotensin III: a potent inhibitor of enkephalin-degrading enzymes and an analgesic agent. AB - Various angiotensins, bradykinins, and related peptides were examined for their inhibitory activity against several enkephalin-degrading enzymes, including an aminopeptidase and a dipeptidyl aminopeptidase, purified from a membrane-bound fraction of monkey brain, and an endopeptidase, purified from the rabbit kidney membrane fraction. Angiotensin derivatives having a basic or neutral amino acid at the N-terminus showed strong inhibition of the aminopeptidase. Dipeptidyl aminopeptidase was inhibited by angiotensins II and III and their derivatives, whereas the endopeptidase was inhibited by angiotensin I and its derivatives. The most potent inhibitor of aminopeptidase and dipeptidyl aminopeptidase was angiotensin III, which completely inhibited the degradation of enkephalin by enzymes in monkey brain or human CSF. The Ki values for angiotensin III against aminopeptidase, dipeptidyl aminopeptidase, endopeptidase, and angiotensin converting enzyme, which degraded enkephalin, were 0.66 X 10(-6), 1.03 X 10(-6), 2.3 X 10(-4), and 1.65 X 10(-6) M, respectively. Angiotensin III potentiated the analgesic activity of Met-enkephalin after intracerebroventricular coadministration to mice in the hot plate test. Angiotensin III itself also displayed analgesic activity in that test. These actions were blocked by the specific opiate antagonist naloxone. PMID- 3037032 TI - Cholecystokinin stimulates dopamine synthesis in synaptosomes by a cyclic AMP dependent mechanism. AB - The effects of different fragments of cholecystokinin (CCK) on dopamine synthesis were studied in synaptosomal preparations from the striatum, substantia nigra, and frontal cortex. In striatal synaptosomes, dopamine synthesis rate measured by dopamine accumulation was 12.5% lower than that measured by 3,4 dihydroxyphenylalanine (DOPA) accumulation; however, K+-accelerated synthesis was the same for both methods. Synthesis rate was independent of exogenous tyrosine levels. In the three regions studied, the combined stimulatory effects of 8-Br cyclic AMP and high K+ were additive. CCK-5, CCK-3, CCK-27-33, and CCK-8 (sulphated) enhanced synthesis, CCK-5 being the most potent fragment. The nonsulphated octapeptide had no effect. In all three regions, CCK-5 and high K+ had an additive effect on dopamine synthesis; CCK-5 and 8-Br-cyclic AMP together produced the same enhancement of synthesis as CCK-5 alone. CCK-5 produced similar dose-dependent increases in dopamine synthesis and cyclic AMP accumulation in striatal synaptosomes, and both effects were blocked by the CCK antagonist proglumide. PMID- 3037033 TI - Modification of the effects of cytotoxic chemotherapy on the immune responses of cancer patients with a nonsteroidal, antiinflammatory drug, piroxicam. A pilot study of the Eastern Cooperative Oncology Group. AB - We have attempted to modify the manner in which chemotherapy influences immune function in lung cancer patients by adding the nonsteroidal, antiinflammatory drug (NSAID) piroxicam to a cytotoxic drug treatment regimen. Eighteen previously untreated patients with lung cancer received cytotoxic chemotherapy with mitomycin-C (10 mg/m2), vinblastine (6 mg/m2), and cis-platinum (40 mg/m2) (MVP) on day 1 of a 21-day treatment cycle; 12/18 patients also received piroxicam (20 mg/day) beginning 10 days prior to MVP and persisting throughout the 21-day treatment cycle. In vitro tests included (a) phytohemagglutinin (PHA) responsiveness in the presence of indomethacin or interleukin-2 (IL-2), (b) natural killer cell (NK) function, and (c) PHA-induced IL-2 synthesis. In the 6 patients treated with MVP alone, depressed PHA reactivity was found pretherapy, which was augmented by indomethacin, but not by IL-2; this was associated with impaired production of IL-2. By days 7-14, PHA reactivity, IL-2 responsiveness, and IL-2 production rebounded to normal. By days 14-21, PHA reactivity had declined once again in association with increased indomethacin-regulation and impaired IL-2 responsiveness and production. In the 12 patients who received MVP plus piroxicam, PHA and IL-2 assays improved by days 7-14 and indomethacin regulation was normal; moreover, this improvement persisted throughout the 21-day treatment cycle. Immunological rebound persisted through as many as 3 consecutive 21-day MVP plus piroxicam treatment cycles in patients who were continuously monitored. In contrast to these results, piroxicam did not alter the way chemotherapy affected NK function. These results demonstrate that the immunomodulatory effects of cytotoxic chemotherapy can be modified or ameliorated by adding an NSAID to the cytotoxic drug treatment regimen; for certain functions, such as mitogen-stimulated blastogenesis or IL-2 production, sustained immunological rebound without immunosuppression can be achieved by this maneuver. PMID- 3037034 TI - Cerebellar metastases: diagnostic and management considerations. AB - Prompted by several unsatisfactory outcomes, we reviewed the records of 59 patients with cerebellar metastases (26 solitary) with respect to clinical presentation, diagnosis, and natural history. Eighty-seven percent of patients initially complained of headache, gait disturbance, and/or dizziness. At time of diagnosis, 92% of patients with solitary cerebellar metastases and 74% of the overall series complained of headache and/or difficulty walking. In three of four cases, magnetic resonance imaging (MRI) was superior to x-ray computed tomography (CT) in detecting the cerebellar lesions. Several patients acutely deteriorated during evaluation or at the initiation of radiation therapy. We conclude that a cancer patient presenting with headache and gait difficulty with or without nausea/vomiting and dizziness should promptly undergo head CT scanning, and that MRI is useful even if CT is negative. In addition, we recommend that patients with documented cerebellar metastases receive high-dose glucocorticoid therapy for 48 to 72 hours before beginning radiation therapy. The presence of symptomatic hydrocephalus or failure to respond to glucocorticoids initially are particularly ominous features that may be best managed by early neurosurgical consultation before beginning radiation therapy. PMID- 3037036 TI - Lymphokine activated killer (LAK) cell mediated killing of human glioma: effect of pretreating glioma with various membrane modifying agents. AB - The killing of human glioma by lymphokine activated killer (LAK) cells was studied. LAK cells generated by culturing recombinant interleukin-2 (IL-2) with human peripheral blood lymphocytes (PBL) obtained from normal volunteers markedly lysed allogeneic glioma grown in tissue culture. Susceptibility of glioma to lysis by LAK cells was abrogated by pretreating the glioma cells with trypsin or chymotrypsin, but was unaffected by pretreatment with hydrocortisone, neuraminidase, glycosidases or sodium periodate. These results suggest that the cell surface determinant on human glioma cells responsible for its tumor selective lysis by LAK is a protein sensitive to trypsin and chymotrypsin. PMID- 3037037 TI - An experimental trial of cyclic nucleotides on multicellular spheroids derived from human brain tumours. AB - The effects of cyclic nucleotides, dibutyryl cyclic adenosine monophosphate and dibutyryl cyclic guanosine monophosphate (db-cAMP and db-cGMP), on the growth rate of multicellular tumour spheroids were evaluated by comparing the growth delay and colony forming efficiency in vitro. Multicellular tumour spheroids were derived directly from human brain tumours. To compare the chemotherapeutic effect of cyclic nucleotides, CCNU was used as a known effective cytotoxic drug on malignant gliomas. Significant growth delay was obtained by db-cAMP (p less than 0.001) while CCNU was tumouricidal rather then producing a delay in growth of the tumour spheroids. Db-cGMP found not to be effective in decreasing the growth rate of the tumour spheroids in vitro (p greater than 0.2). The role of cyclic nucleotides in brain tumours is discussed on a review basis. PMID- 3037038 TI - Physiological characteristics of anterior thalamic nuclei, a group devoid of inputs from reticular thalamic nucleus. AB - This study tested the hypothesis that neurons of thalamic nuclei, which are normally devoid of inputs from the reticular thalamic nucleus, do not display spindle oscillations and related rhythmic spike bursts. This proposal derived from our recent studies indicating that the reticular nucleus is the generator of spindling rhythmicity. We used retrograde tracing methods, intracellular recordings in barbiturized cats, and extracellular recordings of single neurons and field potentials in anteroventral (AV), anteromedial (AM), ventroanterior (VA), ventrolateral (VL), and central lateral (CL) thalamic nuclei in cats with rostral brain stem transections (cerveau isole preparations), before and after administration of barbiturates. The observation that AV and AM nuclei do not receive inputs from the reticular nucleus was confirmed by using injections of horseradish peroxidase conjugated to wheat germ agglutinin confined within the limits of anterior nuclei. Such injections led to massive retrograde labeling in mammillary nuclei and layer VI of the retrosplenial cortex but left free of labeling the neurons of the reticular thalamic nucleus. Intracellular recordings showed that AV-AM neurons discharge tonically in response to a depolarizing current applied at rest, whereas they give rise to a slow spike that underlies a burst of fast action potentials when the membrane is hyperpolarized by 5-12 mV. Despite the fact that they share similar properties with other thalamic neurons, intracellularly recorded AV-AM neurons do not exhibit spindle waves under barbiturate anesthesia, whereas VA-VL, CL, and other thalamocortical neurons that receive afferents from the reticular nucleus commonly display such oscillations. With extracellular recordings performed simultaneously in CL and AV or AM nuclei of the unanesthetized cerveau isole preparation, focal spindle oscillations and related rhythmic high-frequency spike bursts of single CL cells contrasted with absence of spindles and spike bursts in AV or AM neurons. Spindling could be induced in AV-AM nuclei only after administration of barbiturates at doses exceeding 3 mg/kg, and it appeared approximately 35-40 s after the barbiturate effect was detected in the simultaneously recorded CL nucleus. Moreover, the spike bursts that were elicited in AV-AM neurons after barbiturate administration were not temporally related with focal spindles. Since spindle oscillations did not appear intracellularly in AV-AM neurons, the possibility was envisaged that barbiturate-induced spindles were the passive reflection of field potentials actively generated in neighboring thalamic nuclei.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3037035 TI - The correlation of neoplastic vulnerability with central neuroepithelial cytogeny and glioma differentiation. AB - The vulnerability of neuroepithelial cells in the central nervous system (CNS) to neoplastic transformation results from the interaction of several factors: the existence of a reserve population of stem cells, the capability of differentiated cells to reenter the kinetic cycle, the number of replicating cells at risk at a particular time, the length of time during which a particular cell population remains in the cycle, the state of differentiation and the further differentiation potential of that population, and the steps of differentiation that are achieved in successive cell generations. This concept explains many aspects of CNS tumor incidence and the relationship of central neuroepithelial embryonal tumors to tumors of adult cell type. The incidence of different types of central neuroepithelial tumors can be correlated with the width of the window of neoplastic vulnerability. Examples illustrating the existence of only a narrow window include such rare tumors as medulloepitheliomas, cerebral neuroblastomas, gangliogliomas and ependymoblastomas. By contrast, cerebellar medulloblastomas, astrocytomas, mixed astrocytomas and oligodendrogliomas, and glioblastomas exemplify instances in which a relatively wider window of vulnerability exists in the light of cellular neuro-ontogeny and of the capacity of glial cells for postnatal replication. The relationship that may occasionally be established between the development of a glioma and the production of cellular gliosis such as may follow brain injury or accompany multiple sclerosis can also be viewed in the light of that concept. Increasing awareness is needed concerning the development of postradiation gliomas, in particular after the apparently successful treatment of acute lymphocytic leukemia. PMID- 3037039 TI - Single-cell neuronal model for associative learning. AB - Recently, a novel cellular mechanism, activity-dependent neuromodulation, was identified in sensory neurons mediating the gill and tail withdrawal reflexes in Aplysia. This mechanism may explain associative learning on a behavioral level. The present study was designed to mathematically model subcellular events that may underlie this mechanism and to examine the ability of the model to fit available empirical data. In this associative model, the reinforcing or unconditioned stimulus (US) leads to non-specific enhancement of transmitter release from sensory neurons by activating a cAMP cascade. Spike activity in sensory neurons, the conditioned stimulus (CS), transiently elevates intracellular Ca2+. The CS-triggered increases of intracellular Ca2+ "primes" the cyclase and amplifies the US-mediated cAMP synthesis. As a result of pairing specific amplification of cAMP levels, transmitter release is enhanced beyond that produced by unpaired stimuli or by application of the US alone. The model is capable of fitting empirical data on activity-dependent neuromodulation and predicts a characteristic interstimulus interval (ISI) curve. At the subcellular level, the model's ISI function is related to the time course of the buffering of intracellular Ca2+. The magnitude and duration of the pairing specific enhancement of transmitter release is related to the levels and time course of intracellular cAMP stimulation. PMID- 3037041 TI - Resolving GABAA/benzodiazepine receptors: cellular and subcellular localization in the CNS with monoclonal antibodies. AB - Monoclonal antibodies, raised against a purified GABAA/benzodiazepine receptor complex from bovine cerebral cortex, have been used to visualize the cellular and subcellular distribution of receptorlike immunoreactivity in the rat CNS, cat spinal cord, and bovine and postmortem human brain. Two different antibodies have been used for these studies; bd-17 recognizes the beta-subunit (Mr 55 kDa) in all the species tested, whereas bd-24 recognizes the alpha-subunit (Mr 50 kDa) of bovine and human but not rat and cat tissues. In bovine and human brain, both antibodies produced very similar staining patterns, indicating a homogeneous receptor composition, at least in the brain areas investigated. The general distribution and density of receptor antigenic sites in all tissues studied were very similar to that of benzodiazepine binding sites radiolabeled with 3H-Ro 15 1788 and of glutamate decarboxylase (GAD)-stained nerve terminals. The results demonstrate a very high receptor density (around neuronal cell bodies and processes or less discretely distributed) in the rat olfactory bulbs, cerebral cortex, ventral pallidum, islands of Calleja, globus pallidus, hippocampus, dentate gyrus, substantia nigra, geniculate nuclei, inferior colliculus, cerebellum, reticular formation, spinal cord, and retina. In contrast, no receptors could be detected in white matter, pineal, pituitary, adrenals, and superior cervical ganglia. Only among the cerebellar layers did we observe a conspicuous difference between the staining intensity and the radiolabeling. In bovine and postmortem human brain, e.g., hippocampus, dentate gyrus, cerebral cortex, and substantia nigra, the same close correlation between the immunohistochemical and radiohistochemical findings was observed. At the electron microscopic level, the immune reaction product in the rat substantia nigra and globus pallidus, for example, was localized to pre- and postsynaptic membranes of axodendritic and axosomatic synapses. Whether the presynaptic labeling represents GABA autoreceptors is discussed. In the near future, the monoclonal antibodies will be used in double-labeling experiments with GAD to identify those GABAergic projections that are modulated by benzodiazepine minor tranquillizers. Furthermore, they could also be used, in studies of postmortem human brain, to diagnose receptor dysfunction possibly associated with CNS disorders such as epilepsy. PMID- 3037042 TI - Opioid receptor systems and the endorphins: a review of their spinal organization. AB - A review of the spinal organization of opioid receptor systems and endorphins is presented. The review is a consideration of the physiological mechanisms underlying the effect of spinal opioids, the pharmacology of the opioid receptors that moderate a variety of spinal processing systems, and the endorphin systems that act upon the spinal receptors. PMID- 3037040 TI - 4-Aminopyridine produces epileptiform activity in hippocampus and enhances synaptic excitation and inhibition. AB - Using extra- and intracellular recording techniques, we investigated the epileptiform activity induced by low concentrations (5 and 10 microM) of bath applied 4-aminopyridine (4-AP) in the CA3 subfield of rat hippocampal slices. We also studied the effects of 4-AP on the excitatory and inhibitory synaptic conductance changes in CA3 neurons produced by mossy fiber stimulation. Low concentrations of 4-AP induced spontaneously occurring epileptiform discharges at extracellular potassium concentrations between 1 and 10 mM. In contrast, picrotoxin and bicuculline produced spontaneous epileptiform discharges at extracellular potassium concentrations between 5 and 10 mM. The paroxysmal depolarizing shift (PDS) induced by 4-AP was also investigated. At potentials between -40 and -10 mV, the waveform of the PDS consisted of a depolarizing component enveloped by a hyperpolarizing component. The amplitude of the depolarizing component of the PDS was a monotonic function of the membrane potential, and the mean measured reversal potential was -25.7 mV. Under voltage clamp conditions, the measured conductance associated with the depolarizing component of the PDS averaged 110 nS, with a reversal potential of -14.1 mV. Application of 5 microM 4-AP produced an increase in the inhibitory synaptic conductance change calculated from currents measured 15 ms following mossy fiber stimulation. The mean value increased from 35.2 to 58.1 nS (P less than 0.05) without a significant change in reversal potential. A concentration of 10 microM 4-AP also produced an increase in this inhibitory synaptic conductance change (from 53.3 to 66.3 nS, P less than 0.05) but caused a significant depolarization of the reversal potential (from -66.5 to -61.6 mV, P less than 0.05). This change in reversal potential may reflect a prolongation of the excitatory synaptic currents produced by 4-AP that contributes to the current measured 15 ms from the stimulus. Following application of either 5 or 10 microM 4-AP, there were no significant changes in the resting potential or input resistance of the neurons studied. Application of 5 microM 4-AP also significantly increased the amplitude of the measured excitatory synaptic conductance change produced by mossy fiber stimulation (from 27.9 to 44.1 nS, P less than 0.05) without producing a change in the reversal potential. In 5 of 21 neurons studied, a long-lasting outward synaptic current was present at holding potentials near rest following mossy fiber stimulation.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3037043 TI - Radon gas and radon daughters pose potential environmental hazard. PMID- 3037045 TI - Neurenteric cyst diagnosed by technetium-99m pertechnetate sequential scintigraphy. AB - Neurenteric cysts are rare congenital anomalies which present as mediastinal tumors associated with vertebra anomalies. Two-thirds of them are lined with gastric mucosa and are potentially life threatening. An exact differential diagnosis is difficult preoperatively but is absolutely necessary because of the grave prognosis if left untreated. We present a case in which [99mTc]pertechnetate sequential scintigraphy demonstrated gastric mucosa in the cyst and helped to confirm the diagnosis. The scintigraphic findings are correlated with radiologic, sonographic, and pathologic features. PMID- 3037044 TI - Suppression by perchlorate of technetium-99m and iodine-123 secretion in milk of lactating goats. AB - Lactating goats were infused with either technetium-99m (99mTc) or iodine-123 (123I) together with chlorine-36 (36Cl) through an indwelling catheter previously placed in an external pudic mammary artery. The radioisotope infusions were repeated together with 100 mg of sodium perchlorate. There was a rapid transfer of 99mTc and 123I into milk, reaching a peak concentration 30 min after a 15-min infusion. The fractional secretion of 99mTc and 123I in milk was reduced by 70% 80% and 60%-66%, respectively, by perchlorate. The fractional secretion of 36Cl was not affected by perchlorate, and the shape of the 36Cl secretion curve differed from those of 99mTc and 123I, which were similar. It is probable, therefore, that the latter nuclides were secreted by a transport route different from that of chloride. Available data describing the secretion of 99mTc in human milk after pertechnetate administration was reviewed, and it was concluded that perchlorate pretreatment significantly reduced the secretion of 99mTc in human breast milk. PMID- 3037046 TI - Failure of iodine-131 MIBG imaging in small cell lung carcinoma. PMID- 3037048 TI - Bathtub refinishers' lung. PMID- 3037047 TI - Selenium content and distribution of human, cow and goat milk. AB - Studies were undertaken to compare the content and distribution of selenium in human, cow and goat milk. Selenium content of cow milk was found to be lower than that of either human or goat milk. Regardless of source, less than 3% of total milk selenium was associated with the lipid fraction. Selenium within the 120,000 X g supernatant accounted for 72, 62 and 30% of the total in cow, human and goat milk, respectively. Glutathione peroxidase occurred in all milk samples with goat greater than human greater than cow. Percent of total peroxidase activity associated with glutathione peroxidase was 29, 27 and 65 for human, cow and goat milk, respectively. Approximately 20-28% of the selenium in milk was removed by dialysis (molecular exclusion of 6-8 kDa). After gel chromatography, 8-12 selenoprotein fractions were detected in undialyzed skim milk from each species. Most of the glutathione peroxidase activity was found in the fractions corresponding to 170 and 96 kDa in milk from all species examined. The diameric form of glutathione peroxidase also appeared in dialyzed and undialyzed milk. Distinct differences in the content and distribution of selenoproteins among these species in fresh and dialyzed milk are discussed. PMID- 3037049 TI - Evaluation of submandibular gland function by sialo-scintigraphy following sialolithectomy. AB - Submandibular gland function following transoral sialolithectomy was examined by 99mTc-pertechnetate sialo-scintigraphy in 10 cases. An intraindividual comparison between the function of the treated gland and that of the contralateral normal gland was made using a time-activity curve. Although glandular recovery was not affected by the duration of symptoms or the existence of the symptom at mealtimes, it was inversely proportional to the size of the calculus. Furthermore, the prognosis was more favorable in patients when the anatomically normal orifice of the submandibular duct was preserved. PMID- 3037050 TI - Evaluation of a new dense-porous hydroxylapatite endosteal dental implant. AB - A new hydroxylapatite implant with a dense and porous part, improved the implant neck-lamina propria interface when placed in that alveolar bone of dogs. Bone tissue was formed on the surface of the porous part above the ridge of the alveolar bone and fibrous bands from the lamina propria surrounding the neck of the implant were anchored into this newly formed bone. The new tissue structure was very similar to that of natural teeth and gingiva. Quantitative histologic analysis indicated that this tissue arrangement provided protection against bacterial invasion and reduced gingival pocket formation. PMID- 3037051 TI - Evaluation of hydroxylapatite-coated titanium dental implants in dogs. PMID- 3037052 TI - Primary malignant fibrous histiocytoma of the mandible: surgical approach and report of a case. AB - A case of histologically proven malignant fibrous histiocytoma (MFH) of the mandible is presented. This tumor rarely occurs as a primary neoplasm of the mandible. An intraoral block resection of the mandible, including adjacent clinically normal soft tissue was performed. To maintain the soft tissue bed and esthetics, and to preserve mandibular function, immediate reconstruction was carried out with methyl methacrylate, the anatomical configuration of which was derived from the resected specimen. PMID- 3037053 TI - Chlorhexidine: not a drug for all reasons. PMID- 3037054 TI - [Inhibitory effect of retinoid on early antigen induction of Epstein-Barr virus]. PMID- 3037055 TI - Congenital mesoblastic nephroma: a clinicoradiologic study of 17 cases representing the pathologic spectrum of the disease. AB - Congenital mesoblastic nephroma (CMN) is a rare infantile renal tumor with a generally excellent prognosis. We describe 17 tumors that fit into the pathologic spectrum of CMN proposed by Beckwith, which ranges from benign renal tumors, through atypical "gray zone" lesions of more aggressive potential, to "crossover" tumors akin to clear cell sarcoma of kidney. Nine patients with histologically typical CMN were significantly younger and had smaller tumors than did eight patients with atypical CMN. Clinical features did not differ in the two groups of patients. A distinctive "ring sign" on renal sonography was commonly seen in patients with typical intrarenal CMN. All 17 patients were alive with no evidence of disease at a mean follow-up of 10 years. Nephrectomy was adequate therapy for younger infants and for those with typical CMN. Nephrectomy was probably also adequate therapy for infants 3 months of age or younger with atypical CMN, even if the tumor extended to the surgical resection margins and into the perinephric connective tissues. Adjuvant chemotherapy or radiation or both should be reserved for patients older than 3 months who have grossly unresected tumors and for those patients whose tumors have an unequivocally malignant histologic appearance or evidence of aggressive biologic behavior. PMID- 3037056 TI - Uptake, distribution, and oxidation of fatty acids by Leishmania mexicana amastigotes. AB - Fatty acid uptake, distribution, and beta-oxidation were investigated in Leishmania mexicana amastigotes. The uptake of radiolabeled palmitic, stearic, and oleic acids was similar, reaching 3-6 nmol/10(8) cells in 2 min and 8-12 nmol/10(8) cells in 60 min. The percent of radiolabeled fatty acid that was esterified in the form of triglycerides or phospholipids increased from less than 25% at 2 min to 65-86% at 60 min. The dehydrogenase(s) in an amastigote granule fraction were unusual in that the Vmax for long-chain substrates (0.95-1.6 delta Abs units/min-mg protein) approximated the Vmax for short-chain substrates (0.82 2.0 delta Abs units/min-mg protein), and the Km for long-chain substrates was high (approximately 250 microM), in contrast to data for a mammalian liver mitochondrial fraction. The high Vmax and Km for long-chain substrates suggest a biochemical mechanism for the postulated high utilization of fatty acids as an energy source for amastigotes. Although the primary anti-leishmanial agent, Sb in the form of Pentostam, inhibited oxidation of palmitic acid to CO2 by intact organisms, Sb did not significantly inhibit fatty acid uptake or esterification by organisms, or beta-oxidation by the granule fraction, and the mechanism of action of Sb remains unclear. PMID- 3037057 TI - In vitro antileishmanial activity of inhibitors of steroid biosynthesis and combinations of antileishmanial agents. PMID- 3037058 TI - Itraconazole treatment of experimental systemic candidiasis in male rats. AB - After intravenous infection with Candida albicans, rats received daily doses of itraconazole for 3 days. All rats receiving 2.5 mg kg-1 day-1 survived while all rats receiving sham-treatment died. With subsequent reduction of the itraconazole dose to 0.63 mg kg-1 day-1, no survival occurred and mortality rates equalled those of the control group. PMID- 3037059 TI - Molecular basis for the functional abnormality in neutrophils from patients with generalized prepubertal periodontitis. PMID- 3037060 TI - Crevicular fluid collagenase activity in healthy, gingivitis, chronic adult periodontitis and localized juvenile periodontitis patients. PMID- 3037061 TI - The effects on calculus deposits of a dentifrice containing soluble pyrophosphate and sodium fluoride in a Filipino population. PMID- 3037062 TI - Effect of sulfhydryl reagents on tetraethylammonium transport in rat renal brush border membranes. AB - Effect of sulfhydryl reagents on the transport of tetraethylammonium, an organic cation, has been studied in brush border membrane vesicles isolated from rat renal cortex. H+ gradient-dependent uptake of tetraethylammonium by the vesicles was inhibited by various sulfhydryl reagents in a dose-dependent manner, and the potency of the reagents was followed in the order of HgCl2 greater than p chloromercuribenzoate, p-chloromercuribenzene sulfonate (PCMBS) greater than N ethylmaleimide. In the absence of H+ gradient, tetraethylammonium uptake and efflux also were inhibited by p-chloromercuribenzoate and PCMBS. The sulfhydryl reagents did not affect the dissipation rate of H+ gradient across the membranes. Pretreatment of brush border membranes with PCMBS resulted in an inhibition of tetraethylammonium uptake in the presence and absence of H+ gradient, and this inhibition was reversed by subsequent treatment of the vesicles with thiols such as dithiothreitol, glutathione and cysteine. The inhibitory effect by PCMBS pretreatment was protected in the preincubation with unlabeled tetraethylammonium. These results suggest that sulfhydryl reagents inhibit the transport of tetraethylammonium by their specific interaction with the active sites of the carrier, and that sulfhydryl groups are essential for organic cation transport system in renal brush border membranes. PMID- 3037063 TI - Acute and chronic lithium treatments influence agonist and depolarization stimulated inositol phospholipid hydrolysis in rat cerebral cortex. AB - The ability of acute and chronic Li treatment to influence agonist and depolarization-induced phosphoinositide metabolism was examined in rat cerebral cortex slices. After acute treatment (6.75 mEq/kg, 18 hr), [3H]inositol phosphates accumulating in the presence of 100 microM carbachol (muscarinic), 31 mM K+, 300 microM histamine (H1) and 300 microM 5-hydroxytryptamine (5 hydroxytryptamine2) were reduced significantly even after preincubation of slices with 2.5 mM myo-inositol. However, the response to noradrenaline (100 microM) (alpha-1) was unaffected. In the absence of a drug-free period, chronic Li (2 weeks) maintained the reduced phosphoinositide response to receptor agonists and K+, and now even noradrenaline responses were reduced significantly. Dose response curves revealed that reduction in the response to carbachol was due to a fall in maximal response and not in EC50. When rats were withdrawn from chronic treatment for 18 hr, the responses to carbachol were enhanced significantly with respect to untreated controls. Neither acute nor chronic Li treatments altered significantly the overall incorporation of [3H]inositol into phospholipids. Furthermore, Li treatment did not influence the activity of phospholipase C assayed in crude homogenates of cerebral cortex. In conclusion, acute and chronic Li treatments producing less than [1 mM] in cerebral tissue, severely disrupts phosphoinositide metabolism. Although such effects may well be secondary to inhibition of inositol-monophosphatase, they are not reversed by inositol and therefore do not appear to result from depleted phosphoinositides. PMID- 3037064 TI - Characterization of beta-1 and beta-2 adrenoceptors in guinea pig atrium: functional and receptor binding studies. AB - The selective beta-2 adrenoceptor agonist procaterol produced positive inotropic and chronotropic responses over a concentration range of 1 nM to 0.1 mM in spontaneously beating right atria and in three of seven electrically driven left atria. The pD2 values (right atria, 7.30; left atria, 7.18) were midway between its known affinities at beta-1 and beta-2 adrenoceptors and are evidence that positive inotropic and chronotropic responses involve a minor beta-2 adrenoceptor component. The pKB values for procaterol against (-)-isoproterenol in the right atria (5.59) and left atria (5.29) were consistent with its affinity for beta-1 adrenoceptors and suggest that these are responsible primarily for positive inotropic and chronotropic responses. Receptor binding studies in right atrial homogenates showed that [125I]cyanopindolol binding was saturable (KD = 36.2 pM, maximal density of binding sites = 49.2 fmol mg-1 protein) and stereoselective with respect to the isomers of propranolol. Competition binding curves for the beta-1 adrenoceptor antagonist CGP 20712A and beta-2 selective antagonist ICI 118,551 against [125I]cyanopindolol binding were resolved into two components using iterative curve fitting techniques. Binding sites with the characteristics of beta-1 and beta-2 adrenoceptors were present in the proportions of approximately 75 to 25%. These studies indicate either that the beta-1 adrenoceptor is coupled more efficiently to the positive inotropic and chronotropic response than the beta-2 adrenoceptor or that a proportion of the beta-2 adrenoceptors subserve other functions. PMID- 3037065 TI - Autoradiographic localization of beta-1 and beta-2 adrenoceptors in guinea pig atrium and regions of the conducting system. AB - [125I]Cyanopindolol binding to slide mounted guinea pig atrial sections was time dependent (Ki = 2.4 X 10(8) M-1 min-1, K-1 = 1.2 X 10(-3) min-1) saturable (24-32 pM) and stereoselective with respect to the isomers of propranolol. Competition binding curves with the beta-1 selective antagonist CGP 20712A and beta-2 selective antagonist ICI 118,551 revealed the presence of beta-1 and beta-2 adrenoceptors in the proportions of 85 to 15%. X-ray film exposed to sections of guinea pig heart incubated previously with [125I]cyanopindolol with or without ICI 118,551 (70 nM) or CGP 20712A (100 nM) or (-)-propranolol (1 microM) showed a high density and even distribution of beta-1 adrenoceptors and a lower density and even distribution of beta-2 adrenoceptors over the myocardium. Beta-2 adrenoceptors also were located in the epicardium, the adventitia of the pulmonary artery, aorta, superior and inferior vena cava and the endothelium of the superior and inferior vena cava. Development of nuclear emulsion-coated coverslips confirmed these observations and, in addition, localized beta-2 adrenoceptors to nerve tissue and the aortic valve. In the papillary muscle and surrounding Purkinje cells there was a high density and even distribution of beta 1 and a much lower density of beta-2 adrenoceptors. The atrioventricular node, atrioventricular bundle and ventricular Purkinje system also contained both beta 1 and beta-2 adrenoceptors and in comparison with the surrounding myocardium, these tissues had a slightly greater density of beta-2 adrenoceptors. PMID- 3037066 TI - Anatomical evidence for alpha-2 adrenoceptor heterogeneity: differential autoradiographic distributions of [3H]rauwolscine and [3H]idazoxan in rat brain. AB - Previous autoradiographic studies which have localized alpha-2 adrenoceptors within brain have used tritiated derivatives of clonidine and other alpha-2 adrenergic agonists. In the present study we have compared the autoradiographic distributions of binding sites labeled by two reportedly selective alpha-2 adrenoceptor antagonists, [3H]rauwolscine and [3H]idazoxan, in rat brain. The distribution of high affinity [3H]idazoxan binding sites differed markedly from that of high affinity [3H]rauwolscine sites throughout the neuraxis. The distribution of [3H]idazoxan binding sites paralleled closely that of [3H]clonidine sites, and corresponded to areas of noradrenergic innervation. Densest [3H]idazoxan labeling appeared over anterior olfactory nuclei, fundus striatum, septum, thalamus, hypothalamus, amygdala, entorhinal cortex, central gray, inferior colliculus, dorsal parabrachial nucleus, locus ceruleus and nucleus of the solitary tract. In contrast, much lower levels of [3H]rauwolscine labeling appeared over several areas which receive primarily dopaminergic innervation, and thus corresponded closely to [3H]spiroperidol binding distributions. Densest [3H]rauwolscine labeling appeared over nucleus caudate putamen, nucleus accumbens, olfactory tubercle, Islands of Calleja, hippocampus, parasubiculum, basolateral amygdaloid nucleus and substantia nigra. In areas labeled by [3H]rauwolscine, computerized densitometric analysis of autoradiographic saturation curves revealed that the maximum binding values for [3H]idazoxan high affinity binding sites were consistently greater than those for high affinity [3H]rauwolscine sites. The pharmacological characterization of these two binding sites in the accompanying paper supports the present anatomical evidence that [3H]idazoxan labels a heterogenous population of alpha-2 adrenoceptor sites, one population of which is selectively labeled by [3H]rauwolscine. PMID- 3037069 TI - Tissue-dependent alpha adrenoceptor activity of buspirone and related compounds. AB - The present organ chamber and receptor binding studies were designed to evaluate the alpha adrenoceptor subtype (alpha-1 vs. alpha-2) and tissue selectivity of buspirone and the related compounds, gepirone, isapirone and 1-(2-pyrimidinyl) piperazine (a metabolite of buspirone). Buspirone, gepirone and isapirone were found to possess weak alpha-1 adrenoceptor affinity (relative to prazosin) but significant and selective alpha-1 adrenoceptor intrinsic efficacy (relative to norepinephrine, phenylephrine and ST-587), which was expressed in a tissue- and species-dependent manner. The rank order for alpha-1 adrenoceptor affinity (isapirone greater than buspirone approximately equal to gepirone) was different from the rank order for alpha-1 adrenoceptor intrinsic efficacy (buspirone greater than gepirone greater than isapirone). The tissues that were the most responsive to norepinephrine and ST-587 (i.e., rat and rabbit aorta) were the same tissues in which the intrinsic efficacy of buspirone was expressed. In contrast, 1-(2-pyrimidinyl)-piperazine was inactive at alpha-1 adrenoceptors. Although no alpha-2 adrenoceptor intrinsic efficacy was observed for any of the compounds, isapirone and 1-(2-pyrimidinyl)-piperazine displayed weak alpha-2 adrenoceptor affinity (relative to rauwolscine). Recent studies have shown buspirone to have an effect on central and peripheral monoaminergic mechanisms. The demonstration in the present study that buspirone and related compounds display significant alpha adrenoceptor activity suggests that alpha adrenoceptor involvement should be considered as a potential contributing factor in the central nervous system and/or peripheral activity of this class of compounds. PMID- 3037068 TI - Evidence implicating alpha-2 adrenergic receptors in the anticonvulsant action of intranigral muscimol. AB - The effects of systemically administered catecholamine receptor antagonists on the anticonvulsant action produced by local application of a gamma-aminobutyric acid agonist, muscimol, to the substantia nigra of rats were studied. Both electroshock and kindled seizures were studied. Two alpha-2 receptor antagonists, idazoxan and yohimbine, blocked the anticonvulsant action of intranigral muscimol in the electroshock model. Neither a beta nor an alpha-1 adrenergic nor a dopamine-2 receptor antagonist blocked this action. In contrast to electroshock seizures, an alpha-2 antagonist only partially reversed the anticonvulsant action of intranigral muscimol in the kindling model. We interpret the data to indicate that the interaction of norepinephrine with alpha-2 receptors is required for the anticonvulsant action of intranigral muscimol in the electroshock model. The only partial reversal found in the kindling model suggests that nigral control of seizure propagation involves more than alpha-2 receptor mediated neurotransmission. PMID- 3037067 TI - Hypolipidemic effect of polymethylenemethane thiosulfonates: inhibitors of acetoacetyl coenzyme A thiolase. AB - A new series of bifunctional thiosulfonates of the general formula CH3SO2S-(CH2)n SSO2CH3(SS1 polymethylene bis methane thiosulfonate) with variable polymethylene chain lengths (n = 6, 8, 10 and 12) were evaluated for their hypolipidemic action on serum cholesterol and triglyceride levels in rats. Their action was based on their specific inhibitory effect on cytoplasmic acetoacetyl-CoA thiolase, one of the key enzymes in cholesterol biosynthesis. These compounds inhibited the enzyme in vitro and in vivo. The inhibition in vitro was in the order of n = 12 greater than n = 10 greater than n = 8 greater than n = 6 greater than, where n is the number of methylene groups inserted between the two thiosulfonate groups. In vivo, the compounds produced variable hypocholesterolemic and/or hypotriglyceridemic effects when injected into groups of newly weaned rats fed standard chow, high fat or high carbohydrate diets. When the enzyme activity was measured in isolated liver homogenates in vitro after injections of the drugs in vivo, 80% of original thiolase activity was lost. This inhibition of enzyme activity did not seem to be rate limiting for their hypolipidemic action in vivo as these effects did not correlate with the inhibition of the isolated enzyme. The lack of correlation between in vitro and in vivo activity might be due to the compounds affecting other enzyme systems and/or due to their differential disposition in vivo. PMID- 3037070 TI - Hypoxia provokes leukotriene-dependent neutrophil sequestration in perfused rabbit hearts. AB - Isolated rabbit hearts were perfused with salt solution containing autologous 111In-labeled neutrophils to determine whether 1) hypoxia provoked myocardial neutrophil sequestration, 2) neutrophil accumulation could be suppressed by inhibition of lipoxygenase and 3) hypoxic myocardium generated radioimmunoassayable leukotriene B4 (LTB4). Whereas accumulation of 111In-labeled neutrophils by normoxic hearts was minimal, induction of acute myocardial hypoxia by perfusion with hypoxic medium caused a rapid uptake of 111In-labeled neutrophils. Sequestered neutrophils were not released during a subsequent 20-min normoxic perfusion period. Hypoxia-induced neutrophil uptake was prevented by nordihydroguaiaretic acid (NDGA) or diethylcarbamazine (DEC), two structurally different inhibitors of lipoxygenase. Although no radioimmunoassayable LTB4 could be detected in neutrophil-free perfusate from normoxic or hypoxic preparations, hypoxia caused an approximate 2-fold increase in myocardial tissue levels of LTB4. Tissue levels of LTB4 reverted to control values after 20 min of normoxic perfusion. Infusion of exogenous LTB4 also provoked myocardial neutrophil uptake. Viewed collectively, these observations suggest that in isolated buffer-perfused rabbit hearts hypoxia induces LTB4 release from resident myocardial cells, which promotes avid neutrophil sequestration. PMID- 3037071 TI - Characterization of the alpha-1 adrenergic receptors in the thoracic aorta of control and aldosterone hypertensive rats: correlation of radioligand binding with potassium efflux and contraction. AB - Pharmacologic analysis of functional and radioligand binding studies were used to determine whether alterations in adrenergic receptors contribute to the catecholamine supersensitivity observed in the thoracic aorta of aldosterone hypertensive rats (AHR). Adrenergic function was evaluated using receptor mediated contraction and rate of 42K efflux. The pA2 for phentolamine was the same in AHR (7.9 +/- 0.1 and 8.0 +/- 0.1) and control (7.9 +/- 0.2 and 8.1 +/- 0.1) rats whether measured by contraction or 42K efflux. The pA2 value for the selective alpha-1 antagonist prazosin (9.8 +/- 0.1 to 10.7 +/- 0.2) and the alpha 2 antagonist yohimbine (6.6 +/- 0.2 to 7.4 +/- 0.2) was similar in AHR and control groups using both norepinephrine and phenylephrine as agonists. The rank order of potency was prazosin greater than phentolamine greater than yohimbine in both groups. The KD for [3H]prazosin binding to AHR (26 +/- 3 pM) and control tissue (34 +/- 6 pM) agreed with the prazosin KB obtained by measurements of contraction and 42K efflux. The alpha-1 receptor density was also unaltered: 39 +/- 1 fmol/mg in AHR and 44 +/- 3 fmol/mg in control. Assessment of the NE dissociation constant (Ka) by method of fractional receptor inactivation indicated that the Ka was 4.8 X 10(-7) M in both AHR and control tissues. However, at the same receptor occupancy, the NE-induced increase in 42K efflux was elevated markedly in aorta from AHR. We conclude that alteration in adrenergic receptor density, affinity or type is not the cause of catecholamine supersensitivity in the aorta from AHR and that postreceptor events mediate this phenomenon. PMID- 3037073 TI - Effects of noradrenergic denervation on alpha-1 adrenoceptors and receptor stimulated contraction of chick expansor secundariorum muscle. AB - Alpha-1 adrenoceptors were identified in the chick expansor secundariorum (ESM) smooth muscle by ligand binding (using [3H]prazosin) and organ bath techniques. We examined the effects of both reversible (6-hydroxydopamine-induced) and irreversible (surgical) noradrenergic denervation on alpha-1 adrenoceptors in the ESM. We also measured, in vitro, muscle contraction stimulated by the alpha-1 adrenoceptor agonist methoxamine and by 5-hydroxytryptamine. After both surgical and chemical denervation there were decreases in the number of [3H] prazosin binding sites in the ESM. After reversible denervation the decrease in receptor number persisted during a period in which there was extensive reinnervation of the muscle. At a time (7 days after injection) when the ESM was partially reinnervated the maximum methoxamine-stimulated response, but not that of 5 hydroxytryptamine, was reduced. After surgical (but not chemical) denervation there were increases in both protein content and wet weight of the muscle. However, at 7 days after denervation, these effects alone could not account for the observed decrease in receptor number. A small increase in sensitivity (1.4 fold) to methoxamine and a much larger increase in sensitivity (4.5-fold) to 5 hydroxytryptamine developed after surgical denervation but there appeared to be nonspecific decreases in maximum responses to the agonists. It is concluded that both surgical and chemical denervation produced a decrease in alpha-1 adrenoceptor number in the ESM; this may have contributed to the decrease in maximum contractile response. After surgical denervation a nonspecific supersensitivity developed in the ESM; it may be that a nonspecific contribution to methoxamine-stimulated responses was less apparent due to the loss of alpha-1 adrenoceptors. PMID- 3037072 TI - Functional evidence for adenosine A2 receptor regulation of phosphatidylcholine secretion in cultured type II pneumocytes. AB - We examined the effects of P1 purinoceptor agonists on secretion of phosphatidylcholine in primary cultures of rat type II pneumocytes. Adenosine and its nonmetabolizable analogs, 5'-N-ethylcarboxyamidoadenosine (NECA), N6 phenylisopropyladenosine (PIA) and 2-chloroadenosine, increased secretion, and this effect was dependent on concentration. At the highest concentration, all agonists stimulated phosphatidylcholine secretion approximately 2-fold. NECA, with an EC50 of 8.9 X 10(-8) M, was the most potent agonist. The order of potency was NECA greater than 2-chloroadenosine = L-PIA greater than or equal to adenosine greater than D-PIA. The stimulatory effect of 10(-6) M NECA was diminished by the P1 antagonists theophylline and 8-phenyltheophylline. The degree of stimulation by the adenosine analogs as well as its time course was the same as that induced by terbutaline. The effects of the adenosine analogs and of terbutaline were not additive, suggesting a similar mode of action for the beta adrenergic and purinoceptor agonists. Terbutaline and the adenosine analogs increased intracellular cyclic AMP levels. Again, NECA was the most potent adenosine analog. For NECA, L-PIA and adenosine, there was a significant correlation between effects on secretion and on cyclic AMP content. These data suggest that the effect of adenosine on phosphatidylcholine secretion in type II pneumocytes is mediated by the A2 subtype of the P1 purinoceptor. PMID- 3037074 TI - Interactions of diuretics with the peripheral-type benzodiazepine receptor in rat kidney. AB - The peripheral-type benzodiazepine receptor (PBR) has been autoradiographically localized to the thick ascending limb and early distal tubule. To elucidate further the role of this receptor in kidney function, we have examined the effects of all classes of diuretics on the binding of labeled PBR-specific ligands (R05-4864, PK 11195) to rat kidney membranes (13,000 X g X 10 min). Drugs capable of inhibiting R05-4864 binding by 50% at less than 200 microM included: metolazone (IC50 = 1 microM), indacrinone (IC50 = 42 microM), indapamide (IC50 = 58 microM), hydrochlorothiazide (HCTZ; IC50 = 117 microM) and trichloromethiazide (IC50 = 175 microM). Conversely, diuretics of the loop (e.g., furosemide), K+ sparing (e.g., triamterene), and carbonic anhydrase inhibitor (e.g., acetazolamide) classes exerted no significant effects on R05-4864 binding (IC50S greater than or equal to 1 mM). Inhibition by indacrinone was stereoselective. Thiazide-like compounds inhibited R05-4864 binding with a rank-order of potencies similar to that for their enhancement of in vivo natriuresis (metolazone greater than HCTZ approximately equal to trichlormethiazide greater than chlorothiazide). Scatchard analysis revealed that metolazone, indacrinone, indapamide and HCTZ inhibited R05-4864 binding by reducing Kd, with no effect on maximum binding. The apparent Kd of metolazone for the renal PBR was 3.8 X 10(-7) M. IC50 values of 14 metolazone derivatives for inhibition of R05-4864 binding correlated well (r = .71, P less than .01) with their natriuretic efficacies. PK 11195 binding to digitonin (1.2 mg/mg of protein)-solubilized membranes displayed the same rank order of, but was twice as sensitive to inhibition by metolazone, indacrinone, indapamide and HCTZ.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3037075 TI - Dual effects of Haemophilus influenzae on guinea pig tracheal beta-adrenergic receptor function: involvement of oxygen-centered radicals from pulmonary macrophages. AB - The Gram-negative bacterium Haemophilus influenzae has been shown to cause a deterioration of the guinea pig pulmonary beta-adrenergic receptor system. In the present study we investigated further the mechanisms behind this effect. To this extent we evaluated the involvement of pulmonary macrophages (PM). Treatment of guinea pigs with killed H. influenzae bacteria resulted in the accumulation of a factor in the serum which could specifically stimulate PM. Thus stimulated, PM from nontreated animals caused a decrease of tracheal beta-adrenergic receptor function in vitro. This effect was evident by a decrease of the maximal response of the dose-response curves to isoprenaline, whereas the EC50 values did not change. Catalase and thiourea abolished the PM-induced effects, whereas superoxide dismutase did not, indicating that oxygen-centered radicals, in particular the highly reactive hydroxyl radical, may be responsible for the observed effects. In addition, dexamethasone also inhibited the decrease of tracheal beta-adrenergic receptor function. When activated PM, taken from animals that had been pretreated with killed H. influenzae bacteria 4 days beforehand, were stimulated with serum from a H. influenzae-treated animal, a potentiation of tracheal beta-adrenergic receptor function was observed. PMID- 3037076 TI - Changes in the lipid inclusion/Sertoli cell cytoplasm area ratio during the cycle of the human seminiferous epithelium. AB - The area occupied by Sertoli cell lipid inclusions--electron-lucent lipid vacuoles (LLV) and electron-dense lipid droplets (DLD)--at each stage of the cycle of the seminiferous epithelium was measured on electron micrographs in young adults and elderly men, and expressed as the ratio "area occupied by lipid inclusions/area occupied by the Sertoli cell cytoplasm". For LLV this ratio increased from stage I to stage III, and decreased from stage IV to stage VI in young adults. These results suggest that the development of LLV is synchronized with the spermatogenic process: the residual bodies released in stages I and II are phagocytized by Sertoli cells and transformed into LLV; the amounts of LLV decrease in the subsequent stages of the cycle and increase again when new residual bodies appear. In elderly men the ratio LLV/Sertoli cell cytoplasm was 1.9-2.9 times higher than in young adults at each stage of the cycle. This increase may be related to the increased germ-cell degeneration observed in ageing testes, DLD were less abundant than LLV and the DLD/Sertoli cell cytoplasm ratio did not undergo cyclic changes in young adults or elderly men. PMID- 3037077 TI - Involvement of the peripheral nervous system in temporal arteritis-polymyalgia rheumatica. Report of 3 cases and review of the literature. AB - Involvement of the peripheral nervous system is very uncommon in the temporal arteritis-polymyalgia rheumatica syndrome. Three different presentations of the involvement can be recognized: mononeuropathy, polyneuropathy and brachial neuropathy--C-5 radiculopathy. We report 3 patients in whom peripheral nerve symptomatology dominated the clinical picture of the disease, and review 20 previously published cases. PMID- 3037078 TI - N,N-dialkylated leucine enkephalins as potential delta opioid receptor antagonists. AB - A series of N,N-dialkylated leucine enkephalins were prepared in order to study the effect of substitution on antagonist activity at the delta opioid receptor. The target peptides 1-7 were evaluated in the mouse vas deferens (MVD) and guinea pig ileum (GPI) at 1 microM. All of the compounds except [N,N-di-2 phenethyl,Leu5]enkephalin (7) showed antagonist activity in the MVD against the delta receptor agonist [D-Ala2,D-Leu5]enkephalin. The most potent congener, [N,N dibenzyl,Leu5]enkephalin (3), was 2.5-fold more potent than [N,N diallyl,Leu5]enkephalin (1). None of the compounds at 1 microM showed any antagonist activity against agonists for other receptor types. The N,N-di-2 phenethyl (7) and N,N-dioctyl (6) analogues showed significant agonist activity at 1 microM in the MVD. PMID- 3037079 TI - 2'-Fluorinated arabinonucleosides of 5-(2-haloalkyl)uracil: synthesis and antiviral activity. AB - The synthesis of 5-(2-fluoroethyl)-2'-deoxyuridine (FEDU, 4b), its 2'-fluoro analogue 1-(2-deoxy-2-fluoro-beta-D-arabinofuranosyl)-5-(2-fluoroethyl)-1H,3H- pyrimidine-2,4-dione (FEFAU, 4k), and the 2'-fluoro analogue of the potent antiherpes virus compound 5-(2-chloroethyl)-2'-deoxyuridine (CEDU), 5-(2 chloroethyl)-1-(2-deoxy-2-fluoro-beta-D-arabinofuranosyl)-1H,3H-pyr imidine - 2,4 dione (CEFAU, 4i), is described. The antiviral activities of these compounds were determined in cell culture against herpes simplex virus (HSV) types 1 and 2 and varicella zoster virus (VZV). All compounds were shown to possess significant and selective antiviral activity. FEDU proved less potent than CEDU against VZV replication; however, it was more active against HSV-2. CEFAU showed marked activity against HSV-1, HSV-2, and VZV. The compound containing fluorine at both positions, FEFAU, exhibited the strongest antiviral potency against HSV-1, HSV-2, and VZV. It inhibited HSV-1 at a concentration of 0.03-0.2 microgram/mL, HSV-2 at 0.1-0.3 microgram/mL, and VZV at 0.03 microgram/mL. Neither FEDU nor CEFAU or FEFAU exerted a significant inhibitory effect on cell proliferation at a concentration of 100 micrograms/mL. Thus, the cytotoxicity of these compounds is as low as that of CEDU and compares favorably to that of previously described 2' fluoroarabinosyl nucleoside analogues. PMID- 3037080 TI - Radioiodinated p-iodoclonidine: a high-affinity probe for the alpha 2-adrenergic receptor. AB - The chemical synthesis of 2-[(2,6-dichloro-4-iodophenyl)imino]imidazolidine (PIC) and its radioiodinated analogue [125I]PIC is described. PIC was synthesized from 2,6-dichloroaniline in five synthetic steps. This agent displayed a high affinity for the alpha 2-adrenergic receptor (IC50 = 1.5 nM) in competitive binding assays conducted with purified human platelet plasma membrane fractions. For the synthesis of radioiodinated PIC the triazene intermediate 11 was synthesized from 2,6-dichloro-4-nitroaniline in five synthetic steps. Acid-catalyzed decomposition of 11 with no-carrier-added Na125I afforded high specific activity [125I]PIC. In view of its high affinity for the alpha 2-adrenergic receptor, [125I]PIC is a potentially useful probe for studies in adrenergic pharmacology. PMID- 3037081 TI - Synthesis of beta-carboline-benzodiazepine hybrid molecules: use of the known structural requirements for benzodiazepine and beta-carboline binding in designing a novel, high-affinity ligand for the benzodiazepine receptor. AB - Hybrid molecules incorporating pharmacologically important structural features of both 3-carboxy-beta-carbolines and 1,4-benzodiazepines were synthesized, and their affinities for the benzodiazepine receptor were determined in vitro. One of these hybrids, 8,14-dioxo-13,14-dihydro-8H-indolo[3',2':4,5]pyrido[2,1-c] [1,4]benzodiazepine (13), demonstrated high affinity for the receptor, displacing both benzodiazepines (IC50 = 23 nM) and beta-carbolines (IC50 = 47 nM) from their binding sites. Of the compounds synthesized, 13 also most closely satisfied the structural requirements that generally ensure a high affinity of both beta carbolines and benzodiazepines for the receptor (e.g., aromaticity of the beta carboline, presence of a carbonyl at C-3 of the beta-carboline and of a pi 2 region on the benzodiazepine). The hybrids not fulfilling these requirements had no affinity for the receptor. In vivo pharmacological properties of 13 could not be demonstrated because of its metabolic instability and/or its poor transport into the brain. The results are discussed in terms of a possible overlapping of beta-carboline binding sites with those of benzodiazepines on the receptor. PMID- 3037082 TI - Human antibodies to ovarian cancer antigens secreted by lymphoblastoid cell lines. AB - Human antibodies, showing specificity for ovarian cancer, have been produced following Epstein-Barr virus transformation of lymphocytes from involved lymph nodes. Specificity was assessed initially using established ovarian cancer and other cell lines and then confirmed by histological examination, employing immunofluorescent staining of tumour tissue. The lymphocytes have been in continuous culture for more than 9 months and secrete IgG antibodies at a rate of 0.5-5 micrograms/ml/10(6) cells/5 days. PMID- 3037083 TI - Decreased hydroxyl radical generation from polymorphonuclear leucocytes in the presence of D-penicillamine and thiopronine. AB - The effect of D-penicillamine (DP) and thiopronine on the generation of reactive oxygen species (ROS) both by stimulated polymorphonuclear leucocytes (PMNs) and in a cell-free, xanthine-xanthine oxidase system was investigated. Both drugs significantly reduced hydroxyl radical (OH.) generation in the PMN system, however, increasing trends of OH. levels were noticed in a cell-free ROS generating system. Although the opposing effects on ROS levels were verified, these two agents showed a similar behaviour presumably due to their structural similarity. The properties of these agents that affect ROS levels may contribute to their beneficial and toxic actions in inflammatory as well as immunoregulatory processes of rheumatoid arthritis (RA). PMID- 3037084 TI - Clinical evaluation of the brachial plexus: a simplified approach. PMID- 3037085 TI - The human V kappa locus. Characterization of extended immunoglobulin gene regions by cosmid cloning. AB - As part of the ongoing work in our laboratory on the structural organization of the human V kappa locus we screened cosmid libraries with V kappa gene probes and obtained numerous V kappa gene-containing cosmid clones. Several genomic regions of the V kappa locus were reconstructed from overlapping cosmid inserts and were extended by one step of chromosomal walking. The regions that are called Wa, Wb, Oa, Ob and Ob' comprise about 370 kb (10(3) bases) of DNA and contain 24 V kappa genes and pseudogenes. The V kappa genes belong to the three dominant subgroups (V kappa I, V kappa II, V kappa III) and are arranged to form mixed clusters with members of the different subgroups being intermingled with each other. The distances between the genes range from 1 to 15 kb. Three genes of the Wa and Wb regions that were sequenced turned out to be pseudogenes. Terminal parts of the regions Wa and Ob that do not contain V kappa genes of one of the known subgroups may represent extended spacer regions within the V kappa locus. Wa and Wb are duplicated regions located at different positions of the locus. Region Wb was found to comprise inversely repeated sections of at least 14 kb each that contain V kappa genes oriented in opposite polarity. This finding is consistent with inversion-deletion models of V-J joining; it also shows that the V kappa locus contains not only unique and duplicated but also triplicated parts. The data on the W and O regions are discussed together with those on the L regions and on other regions established in our laboratory. Although the picture of the human V kappa locus with, to date, about 70 different non-allelic V kappa genes is still incomplete, some general features with respect to the organization of the genes and the limited duplication of genomic regions have emerged. PMID- 3037086 TI - Isolation of intermediates in the binding of the FLP recombinase of the yeast plasmid 2-micron circle to its target sequence. AB - We describe a method for isolating and characterizing intermediates in the binding of the FLP recombinase, encoded by the yeast plasmid 2-micron circle to its target sequence. On a wild-type substrate, three specific complexes are formed. Footprinting analysis of the gel-purified complexes shows that each complex is the result of a unique FLP-DNA association. On the basis of the behavior of various FLP target sequences in the gel-binding assay, we propose a model describing the steps that lead to the formation of a stable FLP-DNA complex. PMID- 3037087 TI - Gene 3 endonuclease of bacteriophage T7 resolves conformationally branched structures in double-stranded DNA. AB - Gene 3 endonuclease of bacteriophage T7 has been expressed from the cloned gene, purified, and characterized as to its activity on different DNA substrates. Besides its known strong preference for cutting single-stranded DNA rather than double-stranded DNA, the enzyme has a strong preference for cutting conformationally branched structures in double-stranded DNA, either X or Y-shaped branches. Three types of branched DNA substrates were used: relaxed circular DNAs containing large cruciform structures (a model for Holliday structures, presumed intermediates in genetic recombination); X-shaped molecules having a limited potential for branch migration, made from the cloned phage and bacterial arms of the lambda attachment site; and Y-shaped molecules, made by hybridizing molecules homologous except for a 2 X 21 base-pair palindrome in one of them. Gene 3 endonuclease cuts two opposing strands at or near the branchpoint to resolve these substrates into linear molecules, and does not cut the potentially single stranded tips of the stem-and-loop structure generated from the palindrome. The position of the cleavage points on the equivalent arm of two X-shaped molecules, constructed from wild-type and mutant lambda attachment sites, show that the enzyme can cut at several different sites within or slightly 5' of the limited region of branch migration. The various activities of gene 3 endonuclease are consistent with the known role of this enzyme in genetic recombination, in maturation and packaging of T7 DNA, and in degradation of host DNA, and suggest that the enzyme recognizes a specific structural feature in DNA. Its cleavage specificity, ready availability, and ability to act at physiological pH and ionic conditions may make gene 3 endonuclease useful as a probe for specific DNA structures or for binding of proteins that alter DNA structure. PMID- 3037089 TI - Arrhythmogenic and electrophysiological effects of alpha adrenoceptor stimulation during myocardial ischaemia and reperfusion. AB - To examine possible arrhythmogenic effects of alpha adrenoceptor stimulation, we studied the effects of methoxamine 10(-6) M on arrhythmias and cellular electrophysiology during global myocardial ischaemia and reperfusion in isolated Langendorff perfused guinea-pig hearts. To avoid interference from release of endogenous catecholamines during ischaemia or reperfusion, experiments were performed using catecholamine depleted hearts (myocardial noradrenaline = 11% of control). Catecholamine depletion markedly reduced the incidence of VT and VF during ischaemia and reperfusion and perfusion with methoxamine significantly reversed this. This arrhythmogenic effect of methoxamine was only observed during ischaemia or reperfusion, was independent of beta adrenoceptor blockade and H2 receptor blockade but was abolished by alpha adrenoceptor blockade with phentolamine. Catecholamine depletion blunted the ischaemia induced fall in action potential amplitude and Vmax and prolonged action potential duration and refractory period. Perfusion with methoxamine either partially or completely reversed these effects. Thus, alpha adrenoceptor stimulation has little effect on normally perfused myocardium, but may induce VT or VF during ischaemia or reperfusion. PMID- 3037088 TI - A sensitive procedure to compare amino acid sequences. AB - Methods are discussed that provide sensitive criteria for detection of weak sequence homologies. They are based on the Dayhoff relatedness odds amino acid exchange matrix and certain residue physical characteristics. The search procedure uses several residue probe lengths in comparing all possible segments of two protein sequences, and search plots are shown with peak values displayed over the entire search length. Alignments are automatically effected using the highest search matrix values and without the necessity of gap penalties. Tests for significance are derived from actual protein sequences rather than a random shuffling procedure. PMID- 3037091 TI - Accumulation pattern of amino acid substitutions in protein evolution. AB - A simple method for the evolutionary analysis of amino acid sequence data is presented and used to examine whether the number of variable sites (NVS) of a protein is constant during its evolution. The NVSs for hemoglobin and for mitochondrial cytochrome c are each found to be almost constant, and the ratio between the NVSs is close to the ratio between the unit evolutionary periods. This indicates that the substitution rate per variable site is almost uniform for these proteins, as the neutral theory claims. An advantage of the present analysis is that it can be done without knowledge of paleontological divergence times and can be extended to bacterial proteins such as bacterial c-type cytochromes. It is suggested that the NVS of cytochrome c has been almost constant even over the long period (ca. 3.0 billion years) of bacterial evolution but that at least two different substitution rates are necessary to describe the accumulated changes in the sequence. This "two clock" interpretation is consistent with fossil evidence for the appearance times of photosynthetic bacteria and eukaryotes. PMID- 3037092 TI - Identification of a glycine-like fragment on the strychnine molecule. AB - Strychnine, a complex molecule, antagonizes in some unknown manner the action of glycine, an important inhibitory neurotransmitter in the spinal cord and brainstem of many vertebrates. To help understand the mechanism of this antagonism, we have employed modern computational methods to assess the similarities between these seemingly different molecules. An exhaustive comparison of topological and electronic features of both molecules was made. We have successfully located a glycine-like fragment in the strychnine molecule that, when compared to glycine, exhibits both topological and electronic charge congruence. The successful location of this glycine-like fragment allows us to speculate how the large strychnine molecule assumes its role as an antagonist against the inhibitory action of glycine, the simplest amino acid. PMID- 3037090 TI - Insertion of an intermediate repetitive sequence into a sea urchin histone-gene spacer. AB - A common polymorphism of the early embryonic histone-gene repeat of Strongylocentrotus purpuratus is a 195-bp insertion within the H4-H2B spacer. The sequence, found as an insert in histone-gene repeats of 6 of 22 individuals screened, is also found at approximately 50 sites elsewhere in the genome of every individual. We compare the sequences of the histone-gene spacers that do and do not contain the insert. The insert is found not to have transposon-like features, and no sequence in the original spacer has been duplicated to flank the insert. There is, however, a hexanucleotide sequence that is repeated three times at one end of the insert, and the element has inserted between direct repeats of 5 bp that were present in the original spacer. One of the copies found outside the histone gene cluster was cloned and sequenced and is compared with the insert. Again, no transposon-like features are evident. Regions flanking the homologous sequence in this clone were used as hybridization probes in whole genome blots. Results indicate that the 195-bp sequence insert is itself embedded within a larger element that is repeated within the genome. Therefore, only a portion of a larger repetitive sequence has integrated into the histone-gene spacer. The sequence features of the insert, although not typical of mobile elements, may be representative of other illegitimate recombination events. PMID- 3037093 TI - Denervation of serotonergic fibers in the hippocampus induced a trophic factor which enhances the maturation of transplanted serotonergic neurons but not norepinephrine neurons. AB - The trophic effects of specific denervation on the growth and survival of fetal serotonergic (5-HT) or norepinephrinergic (NE) neurons grafted into the hippocampus were assessed by means of two transplantation paradigms. In the first, fetal raphe cells (containing 5-HT neurons) were transplanted into the control hippocampus. In the second, the transplantation was performed 2 weeks after the specific removal of 5-HT afferents to the hippocampus with 5,7 dihydroxytryptamine (5,7-DHT). We found that a month after transplantation, the number of 5-HT immunoreactive neurons was not significantly different between the two experimental paradigms. However, transplanted raphe neurons had 400% more 5 HT synaptosomal high-affinity uptake and 380% higher content of 5-HT in the hippocampus with prior 5,7-DHT lesion than in control hippocampus. Furthermore, immunocytochemistry showed that the transplanted 5-HT neurons had denser processes and varicosities in the hippocampus with lesion than in control hippocampus. The somatic area of the neurons with these denser processes and varicosities was 42% larger than that of control group. A greater 5-HT level could be achieved if transplanted neurons in the control hippocampus were treated with the supernatant extracted from the hippocampus with 5,7-DHT lesion. In contrast, the NE level of the implanted fetal locus ceruleus (containing NE neurons) was not significantly higher in the 5-HT denervated hippocampus than in control hippocampus a month after transplantation. These results suggest that 5 HT denervation in the hippocampus induces a trophic substance which promotes the maturation rather than survival of 5-HT neurons but not NE neurons. PMID- 3037095 TI - Regulation of dihydropyridine calcium antagonist binding sites in the rat hippocampus following neurochemical lesions. AB - The effects of catecholaminergic, cholinergic, serotonergic, and glutaminergic terminal destruction and neurotransmitter depletion on [3H]nitrendipine binding to rat brain membranes were determined using the neurotoxins 6-hydroxydopamine, 5,7-dihydroxytryptamine, and kainic acid and the neurotransmitter-depleting agent reserpine. Following intracisternal injection of 6-hydroxydopamine there were time-dependent increases (14-23%) in the density but not change in the affinity of hippocampal [3H]nitrendipine binding sites. 6-Hydroxydopamine significantly increased [3H]nitrendipine binding in the hippocampus 4 and 10 days following injection. However, no significant change in binding was observed at 16 and 26 days. [3H]Nitrendipine binding in the cerebral cortex, striatum, cerebellum, and brain stem was unaffected by 6-hydroxydopamine. Neither 5,7-dihydroxytryptamine nor kainic acid affected [3H]nitrendipine binding in the hippocampus and cerebral cortex. Acute and chronic reserpinization also did not affect [3H]nitrendipine binding in the hippocampus and cerebral cortex. These results indicate that dihydropyridine calcium antagonist bindings sites in rat brain are subject to brain region-specific regulation following neurochemical lesions and may be present in their largest densities on postsynaptic membranes. PMID- 3037094 TI - Alteration of calmodulin distribution does not accompany dopaminergic supersensitization of the mouse striatum. AB - Membrane-bound calmodulin increases following dopaminergic supersensitization of the rat striatum. To assess the generality of this relationship, mice were treated with two different supersensitization paradigms. Calmodulin levels and subcellular distribution were determined by radioimmunoassay. Chronic haloperidol treatment increased striatal D2 dopamine receptor density by 25% but had no effect on membrane-bound calmodulin levels. Similarly, 6-hydroxy-dopamine (6 OHDA) lesions depleted striatal dopamine content greater than 95% without affecting membrane-bound calmodulin. In contrast, soluble calmodulin levels decreased by 15% in the 6-OHDA-lesioned striatum, suggesting that soluble calmodulin is enriched in presynaptic dopaminergic terminals. We conclude that dopaminergic supersensitization can occur in the mouse striatum in the absence of any change in calmodulin distribution. PMID- 3037096 TI - Cyclic nucleotide phosphodiesterase isozymes in neuroblastoma cells. AB - Adenosine 3',5'-cyclic monophosphate (cAMP) content of neurons is determined not only by the rate of synthesis but also by the rate of hydrolysis by cyclic nucleotide phosphodiesterases. Multiple forms of cyclic nucleotide phosphodiesterase exist in brain and other tissues, and these may be regulated by various hormones and neuromodulators. The present study examines this regulation in a cloned line of neuroblastoma cells (N18TG2). A biphasic Lineweaver-Burk plot of cAMP hydrolysis revealed two Kms approximating 5 and 25 microM. Lineweaver Burk plots of cGMP hydrolysis were linear over a range of 1 microM to 1 mM and exhibited a Km of 37 microM. Neither cAMP nor cGMP competed for hydrolysis of the alternative cyclic nucleotide. No evidence for an allosteric activation of cAMP phosphodiesterase by cGMP was found. Calcium regulation of phosphodiesterase was not found in spite of preparation of the cell extract with several protease inhibitors, and addition of exogenous calmodulin. No effect of calmodulin antagonists (calmidazolium, W7, or trifluoperazine) was observed in vitro or in situ. Growth of the cells in the presence of 200 nM 3,5,3'-triiodothyronine (T3) resulted in an increased hydrolysis of cAMP but of cGMP. This increase was attributed to an increase in Vmax with no change in either high or low Km. This response was blocked by cycloheximide, suggesting that the thyroid hormone effect requires protein synthesis. The thyroid hormone response in neuroblastoma cells is compared with the results of other studies of thyroid hormone effects on phosphodiesterase in other tissues in vivo. PMID- 3037097 TI - Renorrhaphy using knitted polyglycolic acid mesh. AB - Surgical techniques for treatment of penetrating trauma to the kidney have ranged from simple drainage to nephrectomy, depending on the severity of the injury. Based upon successful experience in the surgical management of splenic injuries using knitted polyglycolic acid mesh, we report a case in which absorbable mesh was used to repair a laceration of the kidney. PMID- 3037098 TI - Expression of connective tissue stromal elements in human cholangiocarcinomas. An immunohistochemical and ultrastructural study. AB - The results of the study of ten cholangiocarcinomas from intrahepatic or extrahepatic origins are reported. Using both light microscopic immunohistochemistry and transmission electron microscopy the scirrhous stroma of the tumour showed clear evidence for the production of its collagenous, elastic and possibly other fibrillar elements by the neoplastic cells. Our findings refute the view that the occasional spindle cells (i.e. fibroblasts, myofibroblasts or even smooth muscle cells) could play a major role in the production of such a voluminous amount of the various connective tissue elements. Therefore, it seems reasonable to suggest that the scirrhous stroma of cholangiocarcinomas is produced by the malignant cells similar to those of the breast, oesophagus, stomach and ureter. PMID- 3037099 TI - An immunofluorescent assay for rapid detection of cytomegalovirus. PMID- 3037100 TI - New applications of radionuclide studies in thoracic imaging. AB - Future radionuclide studies in the lung will encompass not only improved evaluation of regional ventilation and perfusion in pulmonary embolism and lung cancer, but should also provide new information about alveolar-capillary permeability, cell kinetics in the lung, and pulmonary metabolism and receptors. With this increase in the scope of pulmonary nuclear medicine must come further refinement of current imaging techniques and development of new radiopharmaceuticals. Pulmonary emission tomography (via single photons or positrons), which reduces or eliminates functional superimposition in the lung, seems to be the most promising technique for performing such studies. In the area of new radiopharmaceuticals, radiolabeled monoclonal antibodies appear promising for studies of lung cancer. These developments suggest that the next few years will be exciting ones for those interested in pulmonary radionuclide studies. PMID- 3037101 TI - Expression of a cellular gene cloned in herpes simplex virus: rabbit beta-globin is regulated as an early viral gene in infected fibroblasts. AB - We constructed nondefective herpes simplex virus type 1 recombinants bearing the intact rabbit beta-globin gene inserted into the viral gene for thymidine kinase to study the expression of a cellular gene when it is present in the viral genome during lytic viral infections. The globin promoter was activated to high levels during productive infection of Vero cells, giving rise to properly spliced and processed cytoplasmic globin transcripts. Expression of globin RNA occurred with early kinetics, was not affected by blocking viral DNA replication, and was strongly inhibited by preventing viral immediate-early protein synthesis with cycloheximide. These results support the hypothesis that temporal control of herpes simplex virus early gene expression is accomplished by mechanisms that are not restricted to viral promoters. In addition, these data show that a cellular transcript can be correctly processed and can accumulate to high levels during viral infection; this indicates that the mechanisms of virally induced shutoff of host RNA accumulation and degradation of host mRNAs do not depend on sequence specific differentiation between host and viral RNAs. These findings also suggest that herpesviruses have considerable potential as high-capacity gene transfer vectors for a variety of applications. PMID- 3037102 TI - Nickel-induced heritable alterations in retroviral transforming gene expression. AB - Determination of the mutagenic effects of carcinogenic nickel compounds has been difficult because, like many metals, nickel is poorly or nonmutagenic in procaryotic mutagenicity assays. We attempted to characterize nickel-induced genetic lesions by assessing the effect of nickel chloride on the conditionally defective expression of the v-mos transforming gene in normal rat kidney cells infected with the Murine sarcoma virus mutant ts110 (MuSVts110) retrovirus. MuSVts110 contains an out-of-frame gag gene-mos gene junction that prevents the expression of the v-mos gene at the nonpermissive temperature (39 degrees C). In MuSVts110-infected cells (6m2 cells) grown at 33 degrees C, however, this defect can be suppressed by a splicing event that restores the mos reading frame, allowing the expression of a gag-mos fusion protein which induces the transformed phenotype. The capacity to splice the viral transcript at 33 degrees C, but not at 39 degrees C, is an intrinsic property of the viral RNA. This property allowed us to target the MuSVts110 genome using a positive selection scheme whereby nickel was used to induce genetic changes which resulted in expression of the transformed phenotype at 39 degrees C. We treated 6m2 cells with NiCl2 and isolated foci consisting of cells which had reverted to the transformed phenotype at 39 degrees C. We found that brief nickel treatment increased the reversion frequency of 6m2 cells grown at 39 degrees C sevenfold over the spontaneous reversion frequency. The nickel-induced revertants displayed the following heritable characteristics: They stably maintained the transformed phenotype at 39 degrees C; unlike the MuSVts110 RNA in 6m2 cells, the nickel-induced revertant viral RNA could be spliced efficiently at 39 degrees C; as a consequence of the enhanced accumulation of spliced viral RNA, the nickel-induced revertants produced substantial amounts of the transforming v-mos protein P85gag-mos at 39 degrees C; the nickel-induced revertant P85gag-mos serine kinase, like the parental 6m2 P85gag-mos kinase, was found to be rapidly inactivated at 39 degrees C; however, in the nickel-induced revertants, overproduction of P85gag-mos allowed the transformed state to be maintained; and even though viral RNA processing was much changed, no rearrangements of the viral DNA in the nickel induced revertant cells were detected by partial restriction analysis. PMID- 3037103 TI - Expression of human papillomavirus types 6b and 16 L1 open reading frames in Escherichia coli: detection of a 56,000-dalton polypeptide containing genus specific (common) antigens. AB - The human papillomavirus (HPV) genome contains two large open reading frames (ORFs), designated L1 and L2. To characterize the antigenic properties of the L1 ORF-encoded proteins, we cloned the L1 ORFs of HPV6b and HPV16 in plasmids, and these were expressed in Escherichia coli. First, the HPV6b DNA, representing 85.2% of the L1 ORF, was cloned in pUC19 and expressed in E. coli JM83 and RB791 as a 160,000-molecular-weight (160K) fusion protein with E. coli beta galactosidase (6bL1/beta-gal). Second, the HPV16 DNA, representing 89.8% of the L1 ORF, was cloned in pKK233-2 and expressed as a 56K protein (16L1) in strain RB791. Both the 6bL1/beta-gal and 16L1 proteins cross-reacted with anti-bovine papillomavirus type 1 (BPV1) antibody raised against disrupted BPV1 particles. An antibody raised against the 6bL1/beta-gal fusion protein reacted with the 16L1 protein and also with native papillomavirus antigens in human genital condyloma and bovine fibropapilloma tissues, as determined by biotin-streptavidin staining. Furthermore, the anti-6bL1/beta-gal antibody recognized a 54K protein which seemed to be a major capsid protein of BPV1 and also a 56K protein of biopsies harboring HPV6 or HPV11. From these results we concluded that the papillomavirus L1 gene product contains genus-specific (common) antigens and that the HPV6 and HPV11 L1 genes specify the 56K capsid protein. PMID- 3037104 TI - In situ molecular hybridization for detection of Aleutian mink disease parvovirus DNA by using strand-specific probes: identification of target cells for viral replication in cell cultures and in mink kits with virus-induced interstitial pneumonia. AB - Strand-specific hybridization probes were utilized in in situ molecular hybridization specifically to localize replicative form DNA of Aleutian mink disease parvovirus (ADV). Throughout in vitro infection, duplex replicative form DNA of ADV was located in the cell nuclei. Single-stranded virion DNA and capsid proteins were present in the nuclei early in infection, but were later translocated to the cytoplasm. In neonatal mink, ADV causes acute interstitial pneumonia, and replicative forms of viral DNA were found predominantly in alveolar type II cells of the lung. Viral DNA was also found in other organs, but strand-specific probes made it possible to show that most of this DNA represented virus sequestration. In addition, glomerular immune complexes containing intact virions were detected, suggesting that ADV virions may have a role in the genesis of ADV-induced glomerulonephritis. PMID- 3037106 TI - Cells expressing herpes simplex virus glycoprotein gC but not gB, gD, or gE are recognized by murine virus-specific cytotoxic T lymphocytes. AB - To determine which viral molecule(s) is recognized by herpes simplex virus (HSV) specific cytotoxic T lymphocytes (CTL), target cells were constructed which express individual HSV glycoproteins. A mouse L cell line, Z4/6, which constitutively expressed high levels of HSV type 2 (HSV-2) gD (gD-2) was isolated and characterized previously (D. C. Johnson and J. R. Smiley, J. Virol. 54:682 689, 1985). Despite the expression of gD on the surface of Z4/6 cells, these cells were not killed by anti-HSV-2 CTL generated following intravaginal infection of syngeneic mice. In contrast, parental Z4 or Z4/6 cells infected with HSV-2 were lysed. Furthermore, unlabeled Z4/6 cells were unable to block the lysis of HSV-2-infected labeled target cells. Cells which express HSV-1 gB (gB-1) were isolated by transfecting L cells with the recombinant plasmid pSV2gBneo, which contains the HSV-1 gB structural sequences and the neomycin resistance gene coupled to the simian virus 40 early promoter and selecting G418-resistant cell lines. One such cell line, Lta/gB15, expressed gB which was detected by immunoprecipitation and at the cell surface by immunofluorescence. Additionally, cells expressing HSV-1 gC (gC-1) or gE (gE-1) were isolated by transfecting Z4 cells, which are L cells expressing ICP4 and ICP47, with either the recombinant plasmid pGE15neo, which contains the gE structural sequences and the neomycin resistance gene, or pDC17, which contains the gC structural gene coupled to the gD-1 promoter. A number of G418-resistant cell lines were isolated which expressed gC-1 or gE-1 at the cell surface. Anti-HSV-1 CTL generated following footpad infection of syngeneic mice were unable to lyse target cells expressing gB-1 or gE-1. In contrast, target cells expressing very low levels of gC-1 were killed as well as HSV-1-infected target cells. Furthermore, infection of gC-1 transformed target cells with wild-type HSV-1 or a strain of HSV-1 that does not express gC did not result in a marked increase in susceptibility to lysis. These results suggest that murine class I major histocompatibility complex-restricted anti-HSV CTL recognize gC-1 but do not recognize gB, gD, or gE as these molecules are expressed in transfected syngeneic target cells. The results are discussed in terms of recent evidence concerning the specificity of antiviral CTL. PMID- 3037107 TI - High-frequency changes in transcriptional activity in polyomavirus-transformed cell lines. AB - We applied the Luria and Delbruck fluctuation test to analyze high-frequency changes in the phenotype of rat cells transformed by a plasmid carrying the polyomavirus middle T (pmt) gene. All of the transformed cell lines analyzed were capable of switching to the normal state with rates ranging from 10(-3) to 10(-2) per cell per generation. Analysis of both middle T antigen and middle T transcripts indicated that the reversion occurred by a mechanism involving a transcriptional block of the pmt locus. Cell lines containing two separate loci reverted with a lower rate, suggesting that phenotypic switching in these cells involved two independent events affecting each locus. The flat revertants mutated to the transformed state with rates in the range of 10(-5) to 5 X 10(-5) per cell per generation. To determine whether changes in pmt expression would affect neighboring sequences, we transfected a hybrid plasmid carrying pmt linked to the neo marker and selected either for morphological transformants or for G418 resistant cells. Although their coordinate regulation was not absolute, both genes were usually subject to the same changes, reflected by loss and reacquisition of transcriptional activity. PMID- 3037105 TI - The 65,000-Mr DNA-binding and virion trans-inducing proteins of herpes simplex virus type 1. AB - The possible identity of the herpes simplex virus type 1 (HSV-1) 65K (65,000-Mr) virion protein which stimulates transcription from immediate-early genes with the HSV-1 65K DNA-binding protein was investigated. The two proteins were found to be distinct by the three separate criteria of immunological reactivity, tryptic peptide fingerprinting, and mobility in two-dimensional gels. Using HSV-1/HSV-2 intertypic recombinants and a serotype-specific antiserum, we located the gene encoding the 65K DNA-binding protein between coordinates 0.574 and 0.682 on the HSV-1 genome. The protein is posttranslationally modified by phosphorylation. In crude extracts of HSV-1-infected cells the 65K trans-inducing protein did not detectably bind to double-stranded calf thymus DNA under the conditions of our assay. PMID- 3037108 TI - Use of lambda gt11 and monoclonal antibodies to map the genes for the six major glycoproteins of equine herpesvirus 1. AB - To localize the genes for the major glycoproteins of equine herpesvirus 1 (EHV 1), a library of the EHV-1 genome was constructed in the lambda gt11 expression vector. Recombinant bacteriophage expressing EHV-1 glycoprotein epitopes as fusion products with beta-galactosidase were detected by immunoscreening with monoclonal antibodies specific for each of six EHV-1 glycoproteins. Seventy-four recombinant lambda gt11 clones reactive with EHV-1 monoclonal antibodies were detected among 4 X 10(5) phage screened. Phage expressing determinants on each of the six EHV-1 glycoproteins were represented in the library. Herpesviral DNA sequences contained in lambda gt11 recombinants expressing epitopes of EHV-1 glycoproteins were used as hybridization probes for mapping insert sequences on the viral genome. Genes for five EHV-1 glycoproteins (gp2, gp10, gp13, gp14, and gp21/22a) mapped to the genome L component; only one EHV-1 glycoprotein (gp17/18) was expressed from the unique S region of the genome where genes of several major glycoproteins of other herpesviruses have been located. Two glycoproteins of EHV 1, gp13 and gp14, mapped to positions colinear with genes of major glycoproteins identified in several other alphaherpesviruses (gC- and gB-like glycoproteins, respectively). The genomic locations of other EHV-1 glycoproteins indicated the existence of major glycoproteins of EHV-1 (gp2, gp10, and gp21/22a) for which no genetic homologs have yet been detected in other herpesviruses. The results confirm the general utility of the lambda gt11 expression system for localizing herpesvirus genes and suggest that the genomic positioning of several high abundance glycoproteins of EHV-1 may be different from that of the prototype alphaherpesvirus, herpes simplex virus. PMID- 3037109 TI - Bovine leukemia virus transcription is controlled by a virus-encoded trans-acting factor and by cis-acting response elements. AB - Bovine leukemia virus (BLV) gene expression is exquisitely regulated at multiple levels, including a transcriptional control effected by virus-encoded trans acting factors and cis-acting target sequences. Like the human T-cell leukemia viruses type I and type II, but unlike other RNA tumor viruses, BLV contains several open reading frames at the 3' end of its genome. A subgenomic mRNA which encodes two overlapping reading frames from this region could produce proteins of 38 and 18 kilodaltons (kDa). A series of cis-trans experiments using transfected virus gene constructs in different combinations revealed that expression of the 38-kDa protein was both necessary and sufficient to activate, in trans, the BLV promoter. This activation was specific for the BLV long terminal repeat, as a variety of related retroviral promoters were not responsive to the expression of the 38-kDa protein p38(XBL). Deletion analysis and construction of chimeric promoters identified a 75-base-pair long terminal repeat region which functions like a p38(XBL)-dependent enhancer element. PMID- 3037110 TI - Restriction of translation of capped mRNA in vitro as a model for poliovirus induced inhibition of host cell protein synthesis: relationship to p220 cleavage. AB - Poliovirus infection of HeLa cells results in a rapid inhibition of host protein synthesis by a mechanism that does not affect the translation of poliovirus RNA. It has been suggested that this virus-induced translational control results from inactivation of the cap-binding protein complex, and it has been shown that the 220-kilodalton component(s) (p220) of the cap-binding protein complex is cleaved in infected HeLa cells to form antigenically related polypeptides of 100 to 130 kilodaltons. We have previously described an activity in infected cells that specifically restricts translation of capped mRNA in rabbit reticulocyte lysates. Here, we describe further refinements and characterization of restriction assay. We determined that the assay is a good in vitro model for study of host cell shutoff by several criteria: (i) translation was inhibited in both instances at the step involving mRNA binding to ribosomes; (ii) translation of capped mRNA was specifically inhibited, whereas translation of poliovirus RNA was not; (iii) restriction activity appeared in infected cells with kinetics which parallel host cell shutoff; and (iv) restriction activity, like the specific inhibition of host translation, appeared in cells infected in the presence of guanidine-HCl. The restricting activity was partially purified from poliovirus-infected cells and was compared with the virus-induced p220 cleavage activity. Both activities copurified through numerous cell fractionation and biochemical fractionation procedures. However, specific restriction of capped mRNA translation in reticulocyte lysates occurred without complete cleavage of the endogenous p220. PMID- 3037112 TI - Herpes simplex virus virion stimulatory protein mRNA leader contains sequence elements which increase both virus-induced transcription and mRNA stability. AB - To investigate the role of 5' noncoding leader sequence of herpes simplex virus type 1 (HSV-1) mRNA in infected cells, the promoter for the 65,000-dalton virion stimulatory protein (VSP), a beta-gamma polypeptide, was introduced into plasmids bearing the chloramphenicol acetyltransferase (cat) gene together with various lengths of adjacent viral leader sequences. Plasmids containing longer lengths of leader sequence gave rise to significantly higher levels of CAT enzyme in transfected cells superinfected with HSV-1. RNase T2 protection assays of CAT mRNA showed that transcription was initiated from an authentic viral cap site in all VSP-CAT constructs and that CAT mRNA levels corresponded to CAT enzyme levels. Use of cis-linked simian virus 40 enhancer sequences demonstrated that the effect was virus specific. Constructs containing 12 and 48 base pairs of the VSP mRNA leader gave HSV infection-induced CAT activities intermediate between those of the leaderless construct and the VSP-(+77)-CAT construct. Actinomycin D chase experiments demonstrated that the longest leader sequences increased hybrid CAT mRNA stability at least twofold in infected cells. Cotransfection experiments with a cosmid bearing four virus-specified transcription factors (ICP4, ICP0, ICP27, and VSP-65K) showed that sequences from -3 to +77, with respect to the viral mRNA cap site, also contained signals responsive to transcriptional activation. PMID- 3037113 TI - Herpes simplex virus type 1 pathogenicity in footpad and ear skin of mice depends on Langerhans cell density, mouse genetics, and virus strain. AB - Skin Langerhans cells have been shown to be very efficient in presenting antigens to T-helper cells and stimulating the immune response. The present study demonstrates their essential role in the control of primary herpetic infections in the skin. Two unrelated stimuli (abrasion and steroids) were shown to cause depletion of the Langerhans cells in the murine epidermis, and both caused enhancement of the virulence of herpes simplex type 1 (HSV-1) in the skin. The Langerhans cell density was found to be lower in the skin of the ear than in the footpad. HSV-1 was consistently more virulent when injected into the ear epidermis than in the footpad. Thus, HSV-1 pathogenicity in mouse skin depends on the mouse age and strain, the virus strain, and the state of the epidermal Langerhans cells. These findings are discussed in relation to the antigen presenting cell function of the Langerhans cells. PMID- 3037111 TI - 5' long terminal repeats of myc-associated proviruses appear structurally intact but are functionally impaired in tumors induced by avian leukosis viruses. AB - B-cell lymphomas induced in chickens infected with avian leukosis viruses are characterized by integration of the virus within the cellular myc locus and alteration of c-myc expression. Although avian leukosis viruses are intact, replication-competent retroviruses, the structures of many myc-associated proviruses are altered by deletions, raising the possibility that proviral defectiveness plays an essential role in oncogenesis. We found that all myc associated proviruses in 21 independent tumors had deletions, which were confined to the viral genome and did not extend into adjacent cellular sequences. Deletions were not random but, in at least 85% of the myc-associated proviruses, involved a region near the 5' end of the proviral genome where elements implicated in control of viral gene expression have been localized. A second class of deletions involved sequences in the 3' half of the viral genome and included the splice acceptor site used in generating viral env mRNA. Both the 5' and 3' long terminal repeats of myc-associated proviruses appeared to be structurally intact in most tumors, although the 5' long terminal repeats were not involved in expression of either U5-myc transcripts or detectable steady state viral RNAs. A complex array of repeated sequence elements surrounded the junctions of the internal deletions in two myc-associated proviruses. The organization of the deleted proviruses was similar to that of deleted unintegrated viral molecules, consistent with a model in which deletions occurred prior to integration. PMID- 3037114 TI - N-terminal deletion in the src gene of Rous sarcoma virus results in synthesis of a 45,000-Mr protein with mitogenic activity. AB - Expression of the v-src gene of Rous sarcoma virus in avian embryo neuroretina cells results in transformation and sustained proliferation of these normally resting cells. Transformed neuroretina cells are also tumorigenic upon inoculation into immunodeficient hosts. We have previously described conditional mutants of Rous sarcoma virus encoding p60v-src proteins which induce proliferation of neuroretina cells in the absence of transformation and tumorigenicity. These results suggest that p60v-src is composed of functionally distinct domains which may interact with multiple cellular targets. In this study, we describe a spontaneous variant of Rous sarcoma virus, subgroup E, which carries a deletion of 278 base pairs in the 5' portion of the v-src gene but which has retained the ability to induce proliferation of quail neuroretina cells. The deleted v-src gene encodes a 45,000-molecular-weight phosphoprotein which contains both phosphoserine and phosphotyrosine, is myristylated, and possesses tyrosine kinase activity indistinguishable from that of wild-type p60v src. Molecular cloning and sequence analysis of the mutant v-src gene have shown that this deletion extends from amino acid 33 to 126 of the wild-type p60v-src. Therefore, this portion of the v-src protein is dispensable for the mitogenic activity of Rous sarcoma virus in neuroretina cells. PMID- 3037115 TI - Rous sarcoma virus mutant dlPA105 induces different transformed phenotypes in quail embryonic fibroblasts and neuroretina cells. AB - dlPA105 is a spontaneous variant of Rous sarcoma virus, subgroup E, which carries a deletion in the N-terminal portion of the v-src gene coding sequence. This virus was isolated on the basis of its ability to induce proliferation of quiescent quail neuroretina cells. The altered v-src gene encodes a phosphoprotein of 45,000 daltons which possesses tyrosine kinase activity. DNA sequencing of the mutant v-src gene has shown that deletion extends from amino acid 33 to 126 of wild-type p60v-src. We investigated the tumorigenic and transforming properties of this mutant virus. dlPA105 induced fibrosarcomas in quails with an incidence identical to that induced by wild-type virus. Quail neuroretina cells infected with the mutant virus were morphologically transformed and formed colonies in soft agar. In contrast, dlPA105 induced only limited morphological alterations in quail fibroblasts and was defective in promoting anchorage-independent growth of these cells. Synthesis and tyrosine kinase activity of the mutant p45v-src were similar in both cell types. These data indicate that the portion of the v-src protein deleted in p45v-src is dispensable for the mitogenic and tumorigenic properties of wild-type p60v-src, whereas it is required for in vitro transformation of fibroblasts. The ability of dlPA105 to induce different transformation phenotypes in quail fibroblasts and quail neuroretina cells is a property unique to this Rous sarcoma virus mutant and provides evidence for the existence of cell-type-specific response to v-src proteins. PMID- 3037116 TI - Processing of vimentin occurs during the early stages of adenovirus infection. AB - Evidence is presented that a fraction of vimentin, a component of cytoskeleton recently found to be associated with intracytoplasmic, migrating adenovirus type 2 (Ad2), is processed into smaller polypeptides at early times after infection. The extent of vimentin cleavage appears to depend upon both the multiplicity of infection and the adenovirus serotype. Ad2, Ad5, Ad4, and Ad9 induced similar vimentin cleavage in infected cells, whereas Ad3, Ad7, and Ad12, for which most infecting particles are found sequestered within phagosomes, induced very little, if any, vimentin breakdown. This suggests that vimentin processing is in some way related to the number of virus particles migrating through the cytoplasm. Experiments performed in vitro and in vivo with adenovirus temperature-sensitive mutants H2 ts1 and H2 ts112 and UV-inactivated wild-type Ad2 indicated that vimentin processing is due to a nonvirion, cytoskeleton-associated, proteolytic enzyme activated by adenovirus and sharing characteristics with the protease described by Nelson and Traub (W.J. Nelson and P. Traub, J. Cell Sci. 57:25-49, 1982). The activity of this protease appears to be required for productive infection by adenovirus serotypes 2 and 5 (subgroup C), 4 (subgroup E), and 9 (subgroup D) but not by the oncogenic serotypes 3 and 7 (subgroup B) and 12 (subgroup A). PMID- 3037117 TI - Murine gamma interferon inhibits v-mos-induced fibroblast transformation via down regulation of retroviral gene expression. AB - Expression of the retroviral vector Neor myeloproliferative sarcoma virus (MPSV), which contains the v-mos oncogene and the neomycin resistance gene, leads to neoplastic transformation of mouse fibroblasts. Murine recombinant gamma interferon (IFN-gamma) could revert the neoplastic properties of established Neor MPSV-transformed cell lines to an apparently untransformed phenotype. In the presence of IFN-gamma, the Neor MPSV transformants showed a greater than 97% reduction of cloning efficiency in soft agar, strongly reduced proliferative capacity, and morphological changes. The IFN-gamma-induced phenotypic reversion was preceded by a rapid and selective reduction of all retroviral RNA species, apparently due to IFN-gamma action on the long terminal repeat of Neor MPSV. The mRNA levels of cellular genes either remained unaffected (beta-actin) or were even enhanced (H-2) in IFN-gamma-treated Neor MPSV-transformed cells. Upon removal of IFN-gamma, retroviral gene expression was fully recovered and a gradual reappearance of the transformed phenotype of these cells within 3 weeks was noted. These data show that IFN-gamma can cause a virtually complete, but reversible, inhibition of v-mos-induced neoplastic properties in transformed fibroblasts by selective down regulation of retroviral RNA levels. PMID- 3037119 TI - Enhancers and trans-acting E2 transcriptional factors of papillomaviruses. AB - The upstream regulatory regions of human papillomavirus (HPV) types 1, 6b, 7, 11, 16, and 18, bovine papillomavirus type 1, and cottontail rabbit papillomavirus were cloned into transcriptional enhancer assay plasmids which carry the simian virus 40 early promoter lacking its own enhancer and the bacterial gene encoding chloramphenicol acetyltransferase (EC 2.3.1.28) (CAT). Enhancer activity, reflected by CAT gene expression, was detected in all of the upstream regulatory regions tested only when the recombinants were cotransfected with plasmids which express an intact E2 open reading frame of HPV types 1 and 11 or bovine papillomavirus type 1. Each E2 protein stimulated the enhancer from the same virus and, to somewhat lesser degrees, also those from the heterologous viruses. Hence, the enhancer and the E2 protein are functionally conserved among papillomaviruses. There was some nonreciprocity in the extent of trans-activation in heterologous E2-enhancer interactions. Primer extension analyses demonstrated that the E2 proteins increased the abundance of CAT gene mRNA. Tandem multiplication of the HPV type 11 enhancer sequence dramatically increased its response to E2 stimulation; this is possibly relevant to the pathogenicity of papillomaviruses. PMID- 3037118 TI - Human papillomavirus types 6 and 11 mRNAs from genital condylomata acuminata. AB - We have identified and mapped a number of RNA species of human papillomavirus types 6 and 11 from condylomata acuminata by the electron microscopic R-loop technique. Each of the early (E)- and late (L)-region open reading frames (ORFs) deduced from the DNA sequences was represented in one or more transcripts. In addition, RNA species that could encode the modulator of DNA replication and the repressor of transcription, functions recently identified in the genetically similar bovine papillomavirus type 1, were also detected. Some ORFs were 5' proximal in one or more transcripts, whereas others were not 5' proximal in any species, suggesting that internal initiation of translation might be required to gain access to these latter ORFs. Virtually all transcripts had their 5' ends located in the E region and were polyadenylated at one of two sites, i.e., at the end of the E region or at the end of the L region. The great majority of the RNAs were derived from the E region of the genome, with one species approximately 50 to 100 times more abundant than the others. For most of the RNAs, the 5' end mapped near nucleotide 700; minor populations had 5' ends near nucleotide 100 or 1200. By correlating our mapping data with the genomic DNA sequences as well as available RNA structures and cDNA sequences of several papillomaviruses, we predict a number of mRNA splice donor and acceptor sites and suggest that the papillomaviruses have sophisticated usage of ORFs through alternative promoters, mRNA splice sites, and polyadenylation sites. PMID- 3037120 TI - Identification of the gene encoding Marek's disease herpesvirus A antigen. AB - The gene encoding the glycoprotein Marek's disease herpesvirus A antigen (MDHV-A) precursor polypeptide pr47 was delineated by using Northern blot (RNA blot) analysis and hybrid selection of its mRNA with cloned MDHV DNA, cell-free translation of the mRNA, and immunoprecipitation of the polypeptide. The resulting piece of DNA with strongly positive hybrid selection results was a 2.2 kilobase-pair (kbp) PvuII-EcoRI restriction fragment localized to the center of the 18.3-kbp MDHV BamHI B fragment of the total virus genome. The localization was specific since no other small restriction subfragment of the larger BamHI B fragment was able to hybrid select significant MDHV-A mRNA and the gene mapped only in the BamHI B fragment of the total virus genome. Northern blot analysis confirmed the localization of the MDHV-A gene on the 2.2-kbp fragment and detected its mRNA as a 1.8-kilobase species, a size consistent with encoding a 47 kilodalton polypeptide. This is the first report of an MDHV gene being mapped to the MDHV viral genome. This opens the way for the use of recombinant DNA technology to study the nature of the gene encoding a secreted virus-specific glycoprotein that could possibly be involved in immunoprevention, immunosuppression, or immunoevasion, immune phenomena known or speculated to be involved in this oncogenic herpesvirus system. PMID- 3037121 TI - Immunoglobulin A monoclonal antibodies protect against Sendai virus. AB - Immunoglobulin A anti-Sendai virus HN protein monoclonal antibodies, generated via a mucosal immunization protocol, were shown to neutralize virus in vitro and, when passively administered to the mouse respiratory tract, to protect against Sendai virus in vivo. Thus, immunoglobulin A antibodies by themselves can protect against respiratory virus infection. PMID- 3037122 TI - Activated Ha-ras can cooperate with defective simian virus 40 in the transformation of nonestablished rat embryo fibroblasts. AB - We studied the ability of activated Ha-ras to cooperate with simian virus 40 (SV40) in the transformation of nonestablished rat embryo fibroblasts. Cotransfection with Ha-ras greatly accelerated the rate of focus induction by wild-type SV40. Moreover, a series of transformation-defective SV40 mutants could be partially complemented by Ha-ras. This was true not only for mutants retaining an intact N-terminal immortalization-competent domain, but also for a nonkaryophilic SV40 mutant. In the latter case, all detectable T antigen was cytoplasmic, indicating that efficient transformation can be achieved through the interaction of two nonnuclear proteins. By employing cell lines derived with various SV40 mutants, it was determined that the ability to complex with p53 depends on the integrity of a relatively large region in the C-terminal half of large T. Finally, we report that nonkaryophilic SV40 large T forms a complex with the major heat shock protein HSP70, and we discuss its possible implications. PMID- 3037123 TI - Effect of adrenergic and cholinergic drugs on the noradrenergic transmission in bladder neck smooth muscle. AB - Strips of bladder neck smooth muscle were isolated from the dog and noradrenaline stores in the peripheral sympathetic nerve terminals were labelled with (-)3H noradrenaline. Transmural electrical stimulation was applied and the released radiolabeled noradrenaline in the superfusate was measured with a liquid scintillation counter. Autonomic drugs were administered in the superfusate to examine their effect on noradrenaline release evoked by electrical stimulation. The general conclusions drawn from the data are that: there are two autonomic receptor systems on sympathetic nerve terminals of bladder neck--alpha-adrenergic receptors and muscarinic receptors, the activation of alpha-adrenergic receptors or muscarinic receptors depresses noradrenaline release and parasympathetic nerve excitement inhibits noradrenaline release in bladder neck. PMID- 3037124 TI - Bilateral testicular germ cell tumors: a report of 20 cases. AB - Of 412 patients with unilateral testicular cancer 20 (4.3 per cent) suffered a second primary germ cell tumor: 1 had a simultaneous bilateral tumor and in the remaining 19 the second tumor was diagnosed after an interval of 2 months to 32 years. Patients with clinical stages III and IV disease were found only in the group with a second tumor. In 5 patients known risk factors for the development of testicular tumors were found and in 2 prior testicular biopsies showed carcinoma in situ. Effective chemotherapy was used more often in the treatment of the second primary tumor. Of the 20 patients 18 (90 per cent) are free of disease after a mean observation of 5.7 years. A long followup of testicular cancer patients with sonographic evaluation of the remaining testis as well as periodic self-examination by the patient is required. PMID- 3037125 TI - Multilocular cystic nephroma in an adult: immunohistochemical study. AB - We report a case of multilocular cystic nephroma (cysts of the kidney). The tumor was enucleated surgically. It was well encapsulated, and composed of numerous cysts and cellular stroma. Microscopically, the stromal cells resembled blastematous cells seen in Wilms tumor. Immunohistochemical studies disclosed that the stromal cells resembled leiomyocytes, since vimentin and desmin were positive, and myoglobin was negative. An ultrastructural study supported these findings. The characteristics of these cells were compatible with those of congenital mesoblastic nephroma, which also is considered a metanephric tumor. PMID- 3037126 TI - Primary malignant fibrous histiocytoma of the kidney: report of a case. AB - We report a case of primary malignant fibrous histiocytoma of the kidney. Primary involvement of the kidney is rare, with only 8 cases reported in the literature. This malignant mesenchymal tumor is indistinguishable clinically and radiologically from renal cell carcinoma. PMID- 3037127 TI - Tumor necrosis factor enhances the in vitro and in vivo efficacy of chemotherapeutic drugs targeted at DNA topoisomerase II in the treatment of murine bladder cancer. AB - Recombinant human tumor necrosis factor (rTNF) is a macrophage secretory protein with antitumor activity. The murine bladder tumor cell line MBT-2 was used to evaluate the in vitro and in vivo antitumor effects of rTNF in combination with chemotherapeutic drugs targeted at DNA topoisomerase II. These drugs, such as adriamycin and etoposide (VP 16), are in widespread use in the treatment of human cancer. The rTNF significantly enhanced the cytotoxic efficacy of the topoisomerase-targeted drugs actinomycin D, adriamycin, etoposide (VP 16) and teniposide (VM 26) against MBT-2 cells in vitro. The rTNF alone had no effect upon the cells in the same assay. When examined in vivo using MBT-2 tumor-bearing C3H/HeJ mice, these same antitumor relationships were seen. The addition of rTNF to actinomycin D or VP 16 resulted in a significant reduction in tumor volume at 20 days compared to untreated animals. Actinomycin D, VP 16 or rTNF treatment alone had no significant effect on 20 day tumor volume. The data provide a reasonable basis for the addition of rTNF to experimental protocols for the treatment of human bladder cancer using topoisomerase-targeted drugs such as adriamycin both intravesically and systemically. These observations may also be relevant to other human cancers currently treated with these drugs. PMID- 3037128 TI - Adenoviral infections in pediatric liver transplant recipients. AB - Over a 5 1/2-year period, 22 of 262 children receiving liver transplants developed adenoviral infections. Five had adenoviral hepatitis in the allograft, caused by serotype 5. All five were treated for rejection, either just before or at the time of infection. Liver biopsy specimens had characteristic histological appearance, and diagnosis of adenoviral infection was confirmed with monoclonal antiadenoviral antibodies, electron microscopy, and by culture of liver tissue. In the remaining 17 patients, adenovirus was isolated from urine, stool, throat secretions, and/or blood samples, but none had any detectable visceral infection. Serotypes 1 and 2 predominated, similar to children not receiving transplants during the same time period. Three of the patients with hepatitis are alive and well; two died of liver failure. Adenoviral hepatitis did not recur in the second allograft of a patient who underwent retransplantation for combined rejection and adenoviral hepatitis, and appears, therefore, not to be a contraindication to retransplantation when liver failure ensues. PMID- 3037130 TI - [Radiotherapy of lung cancer]. PMID- 3037131 TI - [Gastrointestinal polyposis and hereditary large bowel cancer]. AB - Hereditary large bowel cancer is roughly divided into the hereditary gastrointestinal polyposis (HGIP) that is including adenomatosis coli (AC), Peutz Jeghers syndrome and juvenile polyposis, and non-polyposis familial large bowel cancer (NPF) which is including cancer family syndrome and its related conditions. In the HGIP, AC is most frequent and of highest risk of large bowel cancer, and epidemiological method of approach to patients has been well studied. NPF, however, has been insufficiently studied because of lacking of distinctive marker as seen in the polyposis in spite of that it may be actually present much more frequently than AC. In order to extract NPF from the general large bowel cancer group, the marker or criteria for it has been studied. The factors besides family history presently considered to be useful for detecting NPF are multiple cancers, proximal cancer, association of multiple adenoma and young (50 years or younger) onset of the disease. PMID- 3037129 TI - Plasma concentration of atrial natriuretic polypeptide in patients with atrial tachycardia. AB - To investigate the mechanisms of polyuria associated with tachycardia, we measured plasma concentrations of alpha-human atrial natriuretic polypeptide (alpha-hANP) and cGMP in 6 patients with paroxysmal tachycardia. Plasma concentrations of immunoreactive alpha-hANP and cGMP increased by +69% (p less than 0.05) and +100% (p less than 0.05), respectively, during both paroxysmal atrial tachycardia and atrial fibrillation. To examine whether tachycardia per se raises the secretion of alpha-hANP, we also determined plasma concentrations of alpha-hANP and cGMP in 5 patients during rapid atrial pacing. The pacing-induced tachycardia also increased both of the plasma concentrations. Further, the examinations of cardiac and renal functions in patients with complete atrioventricular block during rapid pacing revealed that each of the increases in atrial pressures, urinary sodium excretion and creatinine clearance were in parallel with the change in plasma concentration of alpha-hANP. These results suggest that an increase in plasma concentration of alpha-hANP during paroxysmal tachycardia is mainly due to elevation of atrial pressure and that this increase in alpha-hANP contributes to tachycardia polyuria. PMID- 3037132 TI - [Epidemiology of neuroblastoma]. AB - From the Childhood Cancer Registry in Japan, incidence of neuroblastoma was 1 among 100,000-150,000 children under the age of 15 years, and 25% of them were under 1 year old. But, mass-screening for this tumor in infancy showed incidence of detection of 1:5,000 babies. Even if we presume that all of the tumor cases- if not screened with the risk of developing later--were detected in infancy, this is more than the expected incidence. To explain this, we postulated 4 patterns in the natural growth of this tumor. After the start of mass-screening, the number of the patients with this tumor in much earlier stage and younger age increased and this was the main cause of improvement of the results of treatment. PMID- 3037133 TI - [Interaction of cancer family history and environmental factors in the risk of cancer--with special reference to childhood malignancies]. AB - Association with family history of cancer of each site was examined for each type of childhood malignancies using data of National Childhood Malignancy Registry in Japan as materials (n = 16,555). 2,926 cases were with cancer family history. Family history of same type of malignancy was found significantly in excess for leukemia, a malignant lymphoma, brain tumor, neuroblastoma and retinoblastoma. When observed by cell type, association with family history of leukemia was most striking in acute myeloid leukemia. Median age at first diagnosis of retinoblastoma was 11 months earlier when family history of retinoblastoma existed. Family history of leukemia and history of exposure to prenatal radiation exposure was found to enhance relative risk for childhood leukemia when combined, suggesting existence of genetic environment interaction. Mode of interaction was interpreted as multiplicative. PMID- 3037134 TI - [Amplification of oncogene N-myc in human neuroblastomas]. AB - The basic mechanisms of oncogene activation are gene amplification, increased expression of a single copy gene and somatic mutation, resulting in a genetic product with increased oncogenic potential. Gene amplification appears to be a more common mechanism of oncogene overexpression and its clinical significance is beginning to be appreciated. In human neuroblastomas, N-myc amplification is highly correlated with advanced stages of disease and rapid disease progression, suggesting that the N-myc gene plays a crucial part in determining the degree of malignancy of neuroblastomas. Chromosome aberrations such as double minutes and homogeneously staining region which observed frequently in neuroblastomas are the cytological manifestation of N-myc amplification and its surrounding DNA fragments. PMID- 3037136 TI - [Histological localization of human papillomaviruses and DNA in the tissues of myrmecia]. PMID- 3037135 TI - [Refractory respiratory tract infections. 7. Trends in the development of new chemotherapeutic agents for respiratory tract infections. c. Combinations of beta lactam antibiotics and beta-lactamases inhibitors]. PMID- 3037137 TI - [A huge subcutaneous tumor occurring on the face (malignant eccrine spiradenoma)- histochemical and electron microscopy study]. PMID- 3037138 TI - [Clear cell hidradenoma--a case report and immunohistological study]. PMID- 3037139 TI - [A clinical study of ruptured hepatocellular carcinoma]. PMID- 3037140 TI - [Effects of alpha-human atrial natriuretic peptide on plasma steroids and renal function in the human]. PMID- 3037141 TI - The time course and extracellular Ca2+ involvement of growth hormone (GH) releasing factor-induced GH secretion in perifused dispersed rat pituitary cells. AB - The time course of GH secretion in response to hpGRF and its dependency on the extracellular Ca2+ concentration were studied in perifused dispersed anterior pituitary cells. The onset of GH secretion in response to 1 nM hpGRF was relatively rapid (within 5 s) but removal of hpGRF after 10-min application further increased the rate of secretion (off-response). The threshold and maximum concentrations of hpGRF in stimulatory secretion were 10(-12) and 10(-8) M respectively. Between these two concentrations, the responses showed dose dependency. A reduction in the extracellular Ca2+ concentration to 0.25 mM or to nominally zero reduced hpGRF-induced GH secretion to 64.4% or to 1.9%, respectively, of the control response in the presence of 2.5 mM Ca2+. Two mM Co2+, known as a strong calcium channel blocker, completely suppressed hpGRF induced GH secretion. The removal of Ca2+ from the perifusion buffer immediately after the offset of 1 min-applied 1 nM hpGRF accelerated the falling phase of GH secretion, which is parallel to the decline in [Ca2+]o in the perifusion chamber. Under nominal Ca2+-free conditions, hpGRF produced no increase in GH secretion. However, 10 min after the offset of 1 min-applied hpGRF under Ca2+-free conditions, the introduction of normal buffer containing 2.5 mM Ca2+ substantially restored GH secretion, although after 20 min the introduction of normal buffer produced only a slight increase in GH secretion. In perifusion experiment of 10(6) cells, intracellular cyclic AMP (cAMP) content was raised from the basal value of 4 to 26 pmol by 2-min application of 1 nM hpGRF. After cessation of hpGRF application, cAMP content decreased to 8.7 pmol at 11 min and returned to the basal value by 20 min. The same tendency was observed in Ca2+ free buffer. In conclusion, the extracellular Ca2+ was essential for hpGRF induced GH secretion. This indicates the importance of the influx of Ca2+ in response to hpGRF. The time course of hpGRF-induced rise and fall in cAMP content was roughly parallel to the GH secretion. The possible explanations of the off response and the restoration of GH secretion by reintroducing normal buffer were discussed. PMID- 3037142 TI - Potentiation of vasopressin secretion by footshocks in rats. AB - Effects of footshocks (FS) on antidiuretic hormone (vasopressin, VP) in the plasma were studied in rats. Continuously applied FS of 60 s period with 5 ms pulses at 50 Hz frequency significantly increased VP as well as adrenocorticotrophic hormone (ACTH) in the plasma in a time- and shock intensity dependent manner. Contrarily, the 50 Hz FS of 2 s period as repeated intermittently at every 15 s for over the period of 2, 10, and 30 min were much less effective for increasing plasma VP, whereas these intermittent FS increased plasma ACTH to an extremely high level. During the inter-shock intervals of 13 s between successive two shock periods rats exhibited a "freezing" behavior. Hypertonic saline or urethane injected I.P. immediately after termination of the intermittent FS significantly increased VP as well as ACTH in the plasma. These data clearly indicate that FS potentiate VP secretion and suggest the possibility that emotional stress may suppress the noxious stimuli-induced VP secretion. PMID- 3037143 TI - [Respiratory disease and clinical pharmacology--serum concentration and pharmacological effect of beta-stimulant and theophylline]. PMID- 3037145 TI - [Chemical mediators in exercise-induced asthma]. PMID- 3037144 TI - [Therapeutic efficacy and pharmacokinetics of cisplatin in patients with non small cell lung cancer]. PMID- 3037146 TI - Hyperplastic and neoplastic alterations in the livers of white perch (Morone americana) from the Chesapeake Bay. AB - White perch (Morone americana) sampled from 15 estuaries of the Chesapeake Bay contained a variety of hyperplastic and neoplastic alterations in their livers. The lesions were derived from bile ductular epithelium and/or hepatocytes. The biliary lesions consisted of hyperplasias and adenomas. The hepatocellular lesions consisted of focal populations of altered cells and neoplasms, of clear cell and basophilic morphologic appearance. The hepatocellular lesions were conspicuous in the absence of the decreased amount of copper accumulation. In this respect, the cells comprising these lesions are reminiscent of cells from hepatocellular neoplasia in mice and rats resistant to accumulation of iron. PMID- 3037148 TI - Dietary fiber sources in the United States by demographic group. AB - Major food sources of total dietary fiber are presented for the whole U.S. population and by age, sex, and race. Revised data on fiber content were applied to dietary intake data from the Second National Health and Nutrition Examination Survey, a representative sample of the U.S. population examined in 1976-80. Vegetables are the principal sources of dietary fiber, followed by bread and fruit. Legumes are important population sources and rank first for several age sex-race categories when food groups are broken down into more detailed food items. Breakfast cereals make considerably lesser contributions. Data on fiber intake in several age, sex, and race groups are also presented. PMID- 3037147 TI - Tumorigenicity test of 1,3- and 1,8-dinitropyrene in BALB/c mice. AB - 1,3-Dinitropyrene (DNP) and 1,8-DNP (CAS: 42397-65-9) are very potent mutagens and induce a frameshift-type mutation in the Salmonella test system. Each compound was tested for tumorigenicity in BALB/c mice by sc inoculation of 0.05 mg of the compound once a week for 20 weeks. Tumors developed at the site of injection of 1,8-DNP in 6 of 15 mice up to 60 weeks after the first injection. The incidence of tumors was statistically significant at a P-value of less than .05 but not of less than .01. Therefore, the carcinogenicity of 1,8-DNP in BALB/c mice was concluded to be weaker than that of benzo[a]pyrene [(BP) CAS: 50-32-8], which induced a 100% tumor incidence when it was injected at the same dose as that of 1,8-DNP. No tumors occurred at the injection site in mice given 1,3-DNP. Most of the tumors induced by 1,8-DNP and BP showed histologic features characteristic of malignant fibrous histiocytoma. PMID- 3037149 TI - Responses of serum from breast cancer patients to murine mammary tumor virus: fact or artifact? AB - For determination of whether breast cancer patients possessed specific serological responses to murine mammary tumor virus (MuMTV), IgG-binding levels were monitored by antibody binding to electrophoretically separated viral proteins (Western blotting and immunodetection) and by the enzyme-linked immunosorbent assay (ELISA) against a panel of five structural proteins (gp55, gp34, p28, p18, and p12) purified from milk-borne MuMTV of the RIII isogeneic mouse strain. No significant antibody reactions were found for sera from 30 cancer patients by the immunoblotting assay, and comparative ELISA studies of 111 patients with malignant mastopathies and 122 healthy, age-matched women revealed no significantly increased mean antibody responses against gp55, gp34, p28, or p12 in breast cancer patients as compared to the responses in the control group. Only for p18 was there a significant increase in mean IgG-binding levels in cancer patients. Additional assays of antibody binding to viral antigens were performed by the cellular immunofluorescence test on MuMTV-expressing cells. These studies also failed to demonstrate greater immunoreactivity of sera from patients as opposed to the immunoreactivity of sera from healthy controls. PMID- 3037151 TI - Scavenging effect of butylated hydroxytoluene on the production of free radicals by the reaction of hydrogen peroxide with N-methyl-N'-nitro-N-nitrosoguanidine. AB - The scavenging effects of the antioxidant butylated hydroxytoluene (BHT) on a hydroxyl free radical (.OH) produced in the reaction of H2O2 with N-methyl-N' nitro-N-nitrosoguanidine [(MNNG) CAS: 70-25-7] and on free radicals derived from MNNG (.MNNG) were examined by electron spin resonance with the use of the spin trapping agent 5,5-dimethyl-1-pyrroline-1-oxide (DMPO). BHT was added to the H2O2 MNNG-DMPO system in 50% acetonitrile and exposed to a tungsten-halogen lamp at an intensity of 0.3 mW/cm2 for 3 minutes at 3 degrees C for the first 2 minutes of irradiation. The amounts of .OH and .MNNG in the system decreased and reached constant levels with increase in BHT concentration. The spectrum of the H2O2-MNNG BHT system showed the specific signal of BHT, whereas the spectra of the BHT solution and the H2O2-MNNG system did not show any signal. These findings indicate that BHT scavenged .OH and .MNNG, and in its reaction with them formed a stable free radical. PMID- 3037150 TI - Integrated state of subgenomic fragments of hepatitis B virus DNA in hepatocellular carcinoma from mainland China. AB - Hepatocellular carcinoma (HCC) samples from mainland China were examined for the presence and state of hepatitis B virus (HBV) DNA sequences. HBV DNA was detected by dot-blot hybridization in 13 of 17 cases of HCC from the Shanghai area and in three of six samples from Hangzhou. The HCC cases from Shanghai were then analyzed in more detail. Fifteen of the 17 patients had serologic evidence of past or present infection with HBV (with inadequate information available for the other two), and the 13 HCC samples positive for HBV DNA all came from serologically positive patients. Southern blot analysis showed that the HBV DNA sequences were always integrated in the HCC high-molecular-weight DNA; only one or two viral copies were present per tumor cell, and no common integration site was evident. Hybridization analyses using subgenomic probes of HBV DNA revealed that the tumors seldom retained an entire HBV genome. HBV S-region sequences were always present, X-region sequences were usually represented, and C-region sequences were rarely detectable in virus-positive tumors. A fragment within the HBV DNA X-region, between nucleotides 1441 and 1526, was found to hybridize nonspecifically with cellular DNA; reported sequence data indicated that this fragment would contain approximately 70% guanine + cytosine. Histologic sections were prepared from some of the frozen tissue specimens and stained by an indirect immunoperoxidase technique for hepatitis B surface antigen (HBsAg). Only 1 of 10 HBV DNA-positive samples contained HBsAg in the cytoplasm of tumor cells, although abundant HBsAg was present in adjacent normal cells in all 10 cases. There were no significant differences in histology between HCC that contained HBV DNA sequences and those that were virus negative. These data support the premise that HBV represents a major etiologic factor in the development of HCC in the Shanghai area of China, although the molecular basis of viral involvement remains obscure. PMID- 3037152 TI - Mammary cancer stages in BALB/cV mice: mouse mammary tumor virus expression and virus-host interactions. AB - A unique subline of BALB/c mice, designated "BALB/cV," exhibits an intermediate mammary tumor incidence (47%) and harbors a distinct milk-transmitted mouse mammary tumor virus (MMTV). Virus expression and virus-host interactions were examined during the different stages of mammary tumorigenesis (normal, preneoplastic, and neoplastic) in the BALB/cV system. Protein immunoblot analyses established the presence of correctly processed (BALB/cV)MMTV structural proteins in all types of BALB/cV mammary tissues. Competition enzyme-linked immunosorbent assays demonstrated that cells from each biologic phenotype were capable of supporting high levels of (BALB/cV)MMTV protein expression. However, mammary epithelial cells that spontaneously underwent the inappropriate pathway of squamous metaplasia did not contain detectable levels of (BALB/cV)MMTV structural proteins. Iodination experiments revealed the presence of a 68K env-related protein on the surface of BALB/cV mammary cells. Nevertheless, sera from 40 mice bearing BALB/cV-positive mammary tissues did not contain detectable levels of anti-env antibodies. Metabolic labeling experiments showed that the half-life of transformation-related, host cell protein p53 (approximately equal to 60 min) in the distinct BALB/cV mammary cell populations was similar to that reported for normal mouse 3T3 cells. It appears that p53 is not stabilized by protein interactions involving any MMTV-encoded or MMTV-induced protein in mammary tumor cells. These characteristics of the BALB/cV system are compatible with the hypothesis that MMTV is only one of two or more cooperating factors required to mediate complete mammary transformation. PMID- 3037153 TI - [AIDS--occupational safety in hospitals--which measures should be taken?]. PMID- 3037154 TI - [Criteria for assessing the adequacy of a hypotensive effect by data on the lactate dehydrogenase and cytochrome oxidase isoenzyme spectra of patients with different forms of arterial hypertension]. AB - A study of associations between lactate dehydrogenase and cytochrome oxidase isoenzymes and total cytochrome oxidase activity, on the one hand, and the type of arterial hypertension, the magnitude of blood pressure and its response to treatment, on the other, was carried out in 143 patients with varying stages of essential hypertension and in renal hypertension. The examined parameters were found to be valuable indicators of adequate hypotensive effect and an optimum BP fall. A 20-24% BP drop below the baseline is optimal for patients with renal hypertension. A 30% BP drop is acceptable for patients with stable essential hypertension. Attaining normal BP level is acceptable and desirable for cases of labile essential hypertension. PMID- 3037156 TI - Expression of the alpha-subunit of Na/K-ATPase in renal collecting duct epithelium during development. AB - Aldosterone enhances synthesis and enzyme activity of Na/K-ATPase and has been found to stimulate sodium reabsorption by the renal cortical collecting duct. Moreover, chronic exposure to aldosterone is associated with a remarkable morphological-functional adaptation. This is seen as a magnification in basolateral membrane area of principal cells. In the present paper we investigated the acquistion of Na/K-ATPase alpha-subunit in renal tissue and examined whether aldosterone initiates functional and adaptive changes in cultured collecting duct cells similar to those observed in vivo. Using a monoclonal antibody, immunofluorescence microscopy demonstrated the acquisition of the alpha-subunit of Na/K-ATPase in the developing cortical collecting ducts of the kidney of neonatal rabbits. The mature collecting ducts in the medulla and papilla of the developing kidney were strongly labelled at the basolateral side, while in the cortical portion of the fetal collecting duct adjacent to the embryonic ampullae the immunolabel was found at the apical and the basolateral aspect of the epithelium. However, the embryonic collecting duct ampullae in the outer cortex did not show any reaction with the antibody. In the collecting duct cells cultured for 24 hours the alpha-subunit of Na/K-ATPase was found to be distributed at both the apical and basolateral aspect of the epithelium. After two to 16 days, the immunolabel was strictly found distributed at the basolateral side. Culturing collecting duct cells in the presence of aldosterone (10(-6) M), the hormone modulated the cellular shape of epithelia after five days by infolding the lateral plasma membranes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3037155 TI - The cAMP system in vasopressin-sensitive nephron segments of the vitamin D treated rat. AB - The present study was undertaken to investigate the cAMP system in isolated vasopressin (AVP)-sensitive segments of the hypercalcemic rat. Hypercalcemia was produced by supplementation of diet with dihydrotachysterol, achieving a mean serum calcium of 12.6 mg%. Maximal urinary concentration was only 1982 +/- 119 mOsm/kg H2O in pair, watered hypercalcemic rats when compared to 2478 +/- 93 mOsm/kg H2O in controls (N = 7) (P less than 0.01). Vasopressin stimulated adenylate cyclase activity at concentrations of vasopressin between 10(-9) and 10(-7) M was indistinguishable in the outer medullary collecting duct (OMCD) and inner medullary collecting duct (IMCD) of tubules dissected from hypercalcemic rats or normocalcemic rats. Likewise, in situ cAMP accumulation in response to 10(-7) M AVP was not significantly different in either OMCD or IMCD of hypercalcemic or normocalcemic rats at either isotonic or hypertonic media conditions. In contrast, while 10(-7) M AVP significantly (P less than 0.05) increased cAMP accumulation in the medullary ascending limb (MAL) of normocalcemic rats it failed to do so in the MAL of hypercalcemic rats. This failure to accumulate cAMP appears to be due to impairment in AVP-stimulated adenylate cyclase rather than to enhanced phosphodiesterase activity. A similar decrement in glucagon stimulated adenylate cyclase occurred with 10(-6) M glucagon. The results demonstrate that in chronic hypercalcemia the cAMP system in the OMCT and IMCD of the rat is intact, but the MAL demonstrates abnormal AVP responsiveness due to impaired adenylate cyclase.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3037157 TI - Angiotensin, ACE inhibition and the kidney. Venice, Italy, September 1985. Proceedings. PMID- 3037158 TI - Angiotensin converting enzyme inhibition and the normal kidney. PMID- 3037159 TI - Effects of angiotensin II and of angiotensin converting enzyme inhibition in chronic renal failure. PMID- 3037160 TI - Disposition of enalapril and enalaprilat in renal insufficiency. PMID- 3037162 TI - Nephrotoxicity of angiotensin converting enzyme inhibition. PMID- 3037161 TI - Lowering of arterial blood pressure limits glomerular sclerosis in rats with renal ablation and in experimental diabetes. PMID- 3037163 TI - Glomerular abnormalities in patients receiving angiotensin converting enzyme inhibitor therapy. PMID- 3037165 TI - Renal effects of enalapril in dogs. PMID- 3037164 TI - Angiotensin converting enzyme inhibition and acute tubular necrosis. PMID- 3037166 TI - Renal damage induced in the clipped kidney of one-clip, two-kidney hypertensive rats during normalization of blood pressure by converting enzyme inhibition. PMID- 3037168 TI - Risks of angiotensin converting enzyme inhibition in renal artery stenosis. PMID- 3037167 TI - Effects on angiotensin-converting enzyme inhibition on renal hemodynamics in renal artery stenosis. PMID- 3037170 TI - Renal alpha 2-adrenoceptors as potential targets for converting enzyme inhibitors. PMID- 3037169 TI - Transrenal changes in active and inactive renin and angiotensin II in renal artery stenosis: effects of converting enzyme inhibition. PMID- 3037171 TI - Renin-angiotensin system and renal circulation in clinical congestive heart failure. PMID- 3037172 TI - Converting enzyme inhibitors and renal function in cardiac failure. PMID- 3037174 TI - Human renal angiotensin I converting enzyme and neutral endopeptidase. PMID- 3037173 TI - Heart failure, renal function, and angiotensin converting enzyme inhibitors. AB - In nonazotemic patients we suggest that ACE inhibitors initially depress GFR, perhaps due to a fall in renal blood flow in those patients in whom a substantial fall in blood pressure occurs with the first dose. Autoregulation of renal blood flow apparently remains intact, but autoregulation of the GFR is impaired during ACE inhibition. Our experience is that sodium and water retention usually ensue and plasma volume expands, leading to a temporary restoration of renal blood flow and GFR towards baseline. Serum sodium is usually reduced at this time, weight in increased, and blood pressure may be partly restored. After about 1-2 weeks, natriuresis begins, although sodium balance and weight probably are not restored to normal for 4-8 weeks. As the plasma volume contracts again, blood pressure falls further and the GFR declines. Renal blood flow, however, is maintained by preferential reduction in renovascular resistance. PMID- 3037175 TI - Acute and chronic effects of angiotensin II on the vessels of the split hydronephrotic kidney. PMID- 3037176 TI - [Fibroepithelial tumors of the breast]. PMID- 3037177 TI - [Extracarotid nonchromaffin paragangliomas]. PMID- 3037178 TI - [Single-stage procedure for peptic ulcer of the gastroenteroanastomosis and diffuse polyposis of the large intestine]. PMID- 3037179 TI - Dexamethasone-suppressible hyperaldosteronism: pathophysiology, clinical aspects, and new insights into the pathogenesis. AB - A profile of dexamethasone-suppressible hyperaldosteronism (DSH), a variant of primary aldosteronism, is drawn by reviewing its pathophysiological and clinical aspects. Genetic studies show no HLA linkage and point to an autosomal dominant mode of inheritance, suggesting that the prevalence of this disease has been underestimated in the past. Hypertension, hypokalemia, suppressed renin, and high aldosterone values characterize DSH in the basal state, similar to the other forms of primary aldosteronism, i.e., aldosterone-producing adenoma (APA) or bilateral idiopathic adrenal hyperplasia (IAH). Biochemically DSH and APA can be differentiated from IAH since in both aldosterone does not respond to upright posture, to angiotensin II infusion, and to angiotensin-converting enzyme (ACE) captopril. In contrast, morphologically DSH is similar to IAH, since neither macroscopic nor histologic examinations of the adrenals give evidence of any unilateral abnormality. However, DSH is differentiated from APA and IAH by the hyperresponsiveness of aldosterone to acute ACTH administration as well as by the failure of aldosterone to escape from prolonged ACTH stimulation. The final diagnosis of DSH rests upon the prompt reversal of the features of mineralocorticoid excess by glucocorticoid therapy. In some cases hypertension is unresponsive to dexamethasone and needs alternative treatment. The main pathogenetic hypotheses point to a pituitary and/or an adrenal abnormality, but the intrinsic nature of the disease remains to be elucidated. PMID- 3037180 TI - [Immune status of patients with bronchial cancer]. AB - The proliferation of lymphocytes, the cell-surface markers of mononuclear cells, and the capacity of T lymphocytes to bind sheep red blood cells were studied in 61 healthy volunteers and 72 patients with small-cell carcinoma, adenocarcinoma, and squamous-cell carcinoma of the lung. The mitogen-stimulated proliferation of the lymphocytes against phytohemagglutinin (PHA) was significantly reduced in patients with small-cell carcinoma. The number of T lymphocytes with T3, T4, T8, and T11 receptors was also reduced, to a degree similar to the E-rosetting rates of patients with small-cell carcinoma. The behavior of the lymphocytes of patients with either adeno- or squamous-cell carcinoma was similar to the normal persons. With regard to prognosis, we could not find significant differences between patients with "limited" and those with "extensive disease". PMID- 3037181 TI - Potentiation of the hCRF-induced release of ACTH in man by an opioid antagonist. AB - Administration of synthetic human corticotropin-releasing factor (hCRF; 2 micrograms/kg body weight) to six normal male subjects produced a significant rise in plasma ACTH, followed by an increase in circulating cortisol. Simultaneous treatment with the opioid antagonist naloxone (1.6 mg i.v. bolus, followed by an infusion at a rate of 1.2 mg/h) significantly potentiated the hCRF induced rise in ACTH and enhanced the cortisol response to hCRF. It is suggested that naloxone acts by antagonizing an inhibitory ultra-short-loop feedback effect of coreleased beta-endorphin on pituitary corticotrophs, thereby amplifying the net effect of hCRF, i.e., the release of ACTH. PMID- 3037182 TI - Atrial natriuretic factor (ANF) inhibits arginine vasopressin-stimulated Ca2+ fluxes and cell contraction in vascular smooth muscle cells. AB - Atrial natriuretic factor (ANF) has been shown to be a potent vasodilator in blood vessels preconstricted by a vasopressor. It is, however, still unknown how ANF interferes with the effects of vasoconstrictors at the cellular level. In the present study the effects of ANF on vasopressin-induced Ca2+ fluxes and cell contraction were examined in primary cultures of vascular smooth muscle cells (VSMC) derived from rat aorta. Spontaneous 45Ca uptake was not affected by 10(-8) M ANF. However, ANF blocked the AVP (10(-8) M)-stimulated 45Ca uptake completely (13.04 +/- 1.42 vs 9.80 +/- 0.46 X 10(3) cpm/mg prot, p less than 0.05). ANF also did not change spontaneous 45Ca efflux from VSMC, whereas it inhibited stimulation of 45Ca efflux by AVP that can be observed within 30 sec (2.59 +/- 0.37 vs 1.39 +/- 0.10 X 10(3) cpm/mg prot/30 sec, p less than 0.01). AVP-induced cell contraction could be prevented by previous incubation with 10(-8) M ANF. ANF stimulated production of cyclic guanosine monophosphate (cGMP) in a dose dependent manner from 10(-9) to 10(-6) M. The stable nucleotide analogue 8-bromo cGMP (1 mM) significantly reduced AVP-stimulated 45Ca efflux (p less than 0.05) and 45Ca uptake (p less than 0.05). Ca fluxes and cell contraction were studied in the presence of 10(-4) M methylene blue (MB) which inhibits soluble guanylate cyclase. MB did not affect the inhibitory effects of ANF on AVP-stimulated 45Ca uptake and efflux. In the presence of MB ANF still blocked AVP-induced cell contraction.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3037185 TI - [Various components of food in the regulation of platelet aggregating activity]. PMID- 3037183 TI - Endogenous Na+ transport inhibitor in human hypertension: further biochemical and chemical studies. AB - Endogenous digitalis-like compound(s) (endalin) has(ve) been reported to be involved in some diseases. Endalin activity is increased in plasma and urine of some essential hypertensives, and in Na+-dependent experimental hypertension. The aims of this study are to compare the biological properties of one endalin extracted from urine of hypertensive patients and of normotensive offspring of hypertensive subjects to those of ouabain and to determine the chemical nature of such an urine-derived endalin. The donors were selected on the basis of the highest Na+,K+-ATPase inhibition produced by extract from their 24-h urine. They consisted of 8 hypertensive patients, 21 normotensive subjects with family history of hypertension and 6 normotensive subjects with no known family history of hypertension. Endalin was semi-purified from 500 liters of pooled urine by flash chromatography on RP 18 packing (40 microns) followed by anion exchange chromatography and two HPLCs on RP 18 reversed phase. Endalin was traced by its capability of inhibiting dog kidney Na+,K+-ATPase activity and 3H-ouabain binding to the enzyme, by its cross-reaction with anti-digoxin antibodies and by its natriuretic effect in rat bioassay. The mechanism of Na+,K+-ATPase inhibition by a semi-purified urine-derived endalin and its consequences on Na-transport were studied and compared to those of ouabain. Semi-purified urine-derived endalin was similar to ouabain in that: it reversibly and specifically inhibited Na+,K+ ATPase activity; it inhibited Na+,K+-ATPase non-competitively with ATP; its inhibitory effect was facilitated by Na+; K+ decreased its inhibitory effect on Na+,K+-ATPase.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3037184 TI - The ouabain receptor in animal tissues and its endogenous ligand. AB - Cardiac glycosides bind to the Na+,K+-ATPase and inhibit its activity. Low concentrations (less than 10(-7) M) of ouabain stimulate the activity of Na+,K+ ATPase in whole homogenates of rat brain. The magnitude of this stimulation varies from 5 to 70%. The concentration of ouabain which induces maximal stimulation is also highly variable and ranges between 10(-9) to 10(-7) M. This stimulation may be explained by the presence of an endogenous ouabain-like compound (OLC) in the brain homogenate. Mammalian tissues and body fluids including brain, heart, kidney, plasma, urine and cerebrospinal fluid contain a unidentified OLC. An endogenous OLC was also demonstrated in toad skin and plasma. This compound was purified to homogeneity and identified using UV, NMR and Mass spectroscopies to be 3-hydroxy-14, 15-epoxy-20,22-dienolide glycoside (resibufogenin). Several reports have suggested that unsaturated fatty acids are the ouabain-like regulators of the Na+,K+-ATPase. Furthermore, Saline infusion to WKY rats, which was shown to increase OLC in the plasma causes also an elevation of free fatty acids. Thus, the interaction of fatty acids with several plasma membrane components was studied. Ouabain binding, opiate binding and binding to the beta-adrenergic receptor were all inhibited by micromolar concentrations of the unsaturated fatty acids, linoleic, oleic and arachidonic. Binding to the opiate receptor was inhibited with IC50 of 40-90 microM and binding to beta adrenergic receptor with IC50 of 350-450 microM.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3037186 TI - Poliovirus resistant cells derived from HeLa cells. PMID- 3037187 TI - [The New-Age-Movement. A short overview]. PMID- 3037188 TI - [Clinical aspects, diagnosis and therapy of polyneuropathies]. PMID- 3037190 TI - Intracytoplasmic eosinophilic inclusion bodies in the liver of beagle dogs. PMID- 3037189 TI - Rederivation of MHV and MEV antibody positive mice by cross-fostering and use of the microisolator caging system. AB - This study established the feasibility of rederiving numerous mouse hepatitis virus (MHV) and mouse encephalomyelitis virus (MEV) antibody positive strains of mice using cross fostering techniques and a new caging system, thus permitting introduction of virus antibody free mice into a barrier facility. Serologic status of dams within the nucleus breeding colony was determined, and all mice within the breeding colony were housed in individual Microisolator cages. Specific pathogen free (SPF) foster mothers purchased from a commercial source were determined to have no detectable serum antibody to 11 murine viruses including MEV and MHV. Pups delivered naturally from time pregnant dams were cross fostered onto the SPF foster dams. The procedure of cross fostering was conducted within a positive flow, HEPA-filtered, mass air displacement unit within 24 hours of parturition. The virus status of pups from 49 litters was monitored serologically at weaning and again at 6 weeks of age. All cross fostered litters were serologically negative for antibody to mouse hepatitis virus. Seven of 29 litters were negative for MEV antibody titer using this cross fostering technique. Those litters negative serologically to both MHV and MEV (at 3 and 6 weeks) were transferred to a barrier facility and held in isolation. All rederived mice transferred to the barrier facility remained negative for MHV and MEV when sampled at 12 weeks of age. PMID- 3037192 TI - Source of iron in neutrophil-mediated killing of endothelial cells. AB - Recently we have shown that human neutrophils activated with phorbol ester are cytotoxic for cultured bovine pulmonary artery endothelial cells in an iron dependent manner. By using the ferric iron chelator deferoxamine mesylate, we have now investigated the source of the iron. Pretreatment of neutrophils with deferoxamine mesylate affected neither their production of O2- nor their cytotoxicity for endothelial cells after addition of phorbol ester. However, similar pretreatment of endothelial cells with deferoxamine mesylate, followed by washing of the cells, resulted in a persistent presence of chelator associated with the endothelial cells and high degrees of protection of endothelial cells from cytotoxicity. The protection was dependent on the amount of chelator used and on the duration of exposure of the endothelial cells to the chelator. These data suggest that iron, which plays an important role in oxygen radical-mediated killing of endothelial cells by neutrophils, is derived from the target (endothelial) cells. PMID- 3037193 TI - Passive inhalation of marijuana smoke: urinalysis and room air levels of delta-9 tetrahydrocannabinol. AB - In two separate studies, 5 drug-free male volunteers with a history of marijuana use were passively exposed to the sidestream smoke of 4 and 16 marijuana cigarettes (2.8% delta-9-tetrahydrocannabinol [THC]) for 1 h each day for 6 consecutive days. A third study was similarly performed with 2 marijuana-naive subjects passively exposed to the smoke of 16 marijuana cigarettes. Passive smoke exposure was conducted in a small, unventilated room. Room air levels of THC and CO were monitored frequently. All urine specimens were collected and analyzed by EMIT d.a.u. assay, Abuscreen radioimmunoassay and GC/MS. The studies show that significant amounts of THC were absorbed by all subjects at the higher level of passive smoke exposure (eg., smoke from 16 marijuana cigarettes), resulting in urinary excretion of significant amounts of cannabinoid metabolites. However, it seems improbable that subjects would unknowingly tolerate the noxious smoke conditions produced by this exposure. At the lower level of passive marijuana smoke exposure, specimens tested positive only infrequently or were negative. Room air levels of THC during passive smoke exposure appeared to be the most critical factor in determining whether a subject produced cannabinoid-positive urine specimens. PMID- 3037191 TI - Characterization of human papilloma virus types in condylomata acuminata in children by in situ hybridization. AB - The presence of human papilloma virus (HPV) DNA sequences of types 6, 11, 16, and 18 was determined by in situ hybridization under stringent conditions on archival paraffin-embedded tissue sections in eight condylomata acuminata observed in children below the age of 12 years. Viral sequences were detected in seven of eight cases: all of them contained HPV 6, four contained also HPV 11, and one contained HPV 16 and 18. Papilloma virus common antigen was detected in only three of eight cases, all of them being positive also by in situ hybridization. We conclude that most condylomata acuminata in children are associated with the same types found in anogenital lesions in adults. Since little is known about the long-term significance of genital condylomas in children the identification of the papilloma virus type may prove to be important as a prognostic tool particularly in patients infected with HPV types 16 and 18, thought to have high oncogenic potential. PMID- 3037194 TI - Potentiated hormonal responses in a model of traumatic injury. AB - Although major trauma often involves repeated insults, few studies have examined the endocrine response to repeated injury. In earlier work, we described potentiated responses of circulating adrenocorticotropin (ACTH) and adrenal secretion of cortisol, epinephrine, and norepinephrine to the second of two small hemorrhages separated by 24 hr. To investigate the response of other hormones to repeated hemorrhage and to examine possible hormonal interactions in a shorter, more clinically relevant time frame, we placed chronic adrenal vein cannulas in eight splenectomized, trained dogs. Each animal was bled 10% of measured blood volume with reinfusion of shed blood at 30 min. The hemorrhage was repeated 5 hr later. Initial hemorrhage led to a small but significant increase in adrenal cortisol secretion and circulating ACTH, vasopressin (AVP), angiotensin II (AII), and plasma renin activity (PRA). Although a 5-hr time interval between stimuli is commonly thought to lead to cortisol feedback, the circulating ACTH and adrenal cortisol secretory responses to the second hemorrhage were exaggerated in comparison to their initial responses. Similarly, the AVP response to the second hemorrhage was also increased. In contrast, the responses of PRA and AII to the second hemorrhage were not greater than their responses to the initial hemorrhage. No differences in measured blood volume before each hemorrhage or in the heart rate and arterial pressure responses to hemorrhage were observed that could explain the potentiated hormonal responses. Thus, potentiated responses of cortisol, ACTH, and AVP, but not of PRA and AII, were observed in a model of trauma that emphasizes repeated injury in a clinically relevant time frame.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3037195 TI - Lung cancer classification: the relationship of disease extent and cell type to survival in a clinical trials population. AB - The staging and histologic cell type of patients in the Lung Cancer Study Group (LCSG) clinical trials program are reviewed and confirmed or resolved at the reference center for anatomic and pathologic classification of lung cancer. A high level of consistency in classification has been achieved through the use of criteria that minimize intraobserver variability. The data obtained from the review project have been used to characterize the relationship of disease extent and cell type to survival in the clinical trials population. Survival characteristics were generated for 1,121 patients who underwent apparent complete resection of nonsmall cell lung cancer and were subsequently entered into various protocols to receive either adjuvant treatment or no further therapy. The end results study provides some insight regarding the biological behavior of squamous cell carcinoma and adenocarcinoma of the lung in terms of the anatomic extent of disease at the time of apparent complete resection. Patients with squamous cell carcinoma had an outcome superior to that of patients with adenocarcinoma in every TNM subset. The differences in survival according to these major cell types were significant overall and in the T1 N0, T1 N1, and T2 N1 subsets but not in the TNM subsets in stage III disease. Histologic cell type and extent of disease are important factors in survival expectations; thus the accuracy and reproducibility of these classifications plays a significant role in the evaluation of differing modalities of treatment. PMID- 3037196 TI - Malignant peripheral primitive neuroectodermal tumor of the uterus. AB - Peripheral primitive neuroectodermal tumor (PPNET) is rare, occurring most often in young adults. Approximately 50 cases have been reported, with only four cases involving the female genital tract. We report the fifth patient. The term "PPNET" should be used only for tumors with neuroectodermal elements exclusively that occur at sites outside the central and sympathetic nervous system. The pathologic differential diagnoses include rhabdomyosarcoma, immature malignant teratoma, small cell carcinoma of the cervix, and ganglioneuroma. Therapy for this tumor has varied, and no effective regimen has been established. PMID- 3037197 TI - Metastatic renal cell carcinoma simulating glomus jugulare tumor. AB - A 59-year-old man presented with jugular foramen syndrome caused by a mass with roentgenographic and histologic features highly suggestive of a glomus jugulare tumor. However, electron microscopic examination of the surgical specimen revealed features diagnostic of a previously unsuspected renal cell carcinoma. Because primary tumors of the glomus jugulare and metastatic renal cell carcinoma may present with the same clinical and roentgenographic findings and look similar histologically, careful electron microscopic examination of the tumor and urologic screening should be performed in suspected cases of glomus jugulare tumors. PMID- 3037198 TI - Recognition of reversible and irreversible myocardial injury by technetium pyrophosphate extraction kinetics. AB - The need for a more accurate method of detecting episodes of myocardial ischemia during cardiac operations, particularly during the ischemic arrest interval, prompted us to investigate the usefulness of measuring the active extraction of technetium pyrophosphate in identifying and quantitating ischemic injury. Twenty four adult mongrel dogs were subjected to cardiopulmonary bypass, and normothermic global ischemia was induced by cross-clamping the proximal aorta. Technetium pyrophosphate (1 mCi) was injected through a standard cardioplegia line with normal saline, simulating administration of cardioplegic solution, upon placement of the aortic cross-clamp (time 0), at 15, 30, 45, and 60 minutes of global ischemia, and with the onset and completion of ischemic contracture. Radioactive counts were recorded over the heart at 1 second intervals, and the extraction fraction and half-time of clearance were calculated. The extraction fraction increased from 0.22 at time 0 to 0.58 at 15 minutes, 0.82 at 30 minutes, 0.85 at 45 minutes, and 0.91 at 60 minutes. The halftime increased from a baseline of 114 seconds (time 0) to a maximum of 321 seconds at 60 minutes of ischemia. The onset and completion of ischemic contracture showed a return toward baseline of both the extraction fraction and halftime of clearance, with an extraction fraction of 0.44 and 0.46 and a halftime of 135 and 133 seconds, respectively. These data clearly show that reversible myocardial injury increased the extraction and reduced the clearance of technetium pyrophosphate and that the magnitude of change related to the extent of injury. The progression to irreversible myocardial injury decreased the active extraction of technetium pyrophosphate. This simple procedure for real-time documentation of myocardial injury promises to provide easily obtainable endpoints of injury for use during cardiac operations in humans. PMID- 3037199 TI - Biochemical and immunological studies on treatment of asthmatic children by means of propagated sensation along channels elicited by meditation. PMID- 3037200 TI - Synaptic connections between neurons in living slices of the larval tiger salamander retina. AB - Synaptic connections between retinal cells were studied by recording simultaneously from pre- and postsynaptic cells in the retinal slice preparation. The time course and waveform of the pre- and postsynaptic light responses were monitored, and the postsynaptic voltage responses to presynaptic current injection were measured. Results obtained provide direct evidence showing that the rod-HBC and rod-HC synapses are sign-preserving, and the rod-DBC, HC-HBC and amacrine-DBC synapses are sign-inverting. Moreover, the synaptic delays between rods and bipolar cells are shorter than that between rods and horizontal cells. The methodology of making retinal slices and the subsequent recording and stimulation procedures are described. The advantages of the retinal slice preparation and its potential in retinal research are discussed. PMID- 3037201 TI - Techniques for obtaining analytical solutions for Rall's model neuron. AB - The Green's function (G) is obtained for a cable equation with a lumped soma boundary condition at x = 0 and a sealed end at x = L infinity. The coefficients in the eigenfunction expansion of G are obtained using the calculus of residues. This expansion converges rapidly for large t. From an estimate of the higher eigenvalues, an approximate bound is obtained for the remainder after so many terms. The leading terms are also obtained in an expansion for G which converges rapidly for small t. Similarly, series expansions for the voltage are obtained which converge rapidly at small or large t when a constant current is injected at the soma and when a (synaptic input) current bte-at occurs at a point along the cable. PMID- 3037203 TI - Influence of age on mitochondrial enzyme levels in Drosophila. AB - The specific activity and the activity per fly of four mitochondrial enzymes did not change with ageing in male Drosophila melanogaster (Oregon R). The enzymes assayed were rotenone-insensitive NADH-cytochrome c reductase, adenylate kinase, succinate cytochrome c reductase, and malate dehydrogenase, located in the outer membrane, inner membrane space, inner membrane and matrix, respectively. The specific activity of malate dehydrogenase showed no significant change for young and old head, thorax and abdomen. We conclude that there is no specific site for ageing damage in the mitochondrion, when the enzyme activities in this study are used as an indicator. It should be noted, however, that these enzymes represent only a small percentage of the total enzymes present in mitochondria. PMID- 3037202 TI - Vascular endothelial markers of the human thoracic duct and lacteal. AB - Factor VIII-related antigen (F8) and Ulex europaeus lectin (UEL) are accepted markers for human blood vessel endothelium. However, disagreement exists as to whether lymphatic vessels stain for F8, and accordingly this study was undertaken to address this issue. Moreover, another vascular endothelial marker, angiotensin converting enzyme (ACE) was also examined in lymphatics. Segments of human thoracic duct and portions of small bowel containing lacteals with post-mortem intervals of less than 15 hours, were removed at autopsy and fixed in B5 or formalin. The specimens were processed routinely and sections examined by indirect immunohistochemical techniques for F8 (Dako Corp.), ACE and for UEL (EY Lab). F8, UEL, and ACE positivity was uniformly found in thoracic ducts and lacteals; however, the staining intensity was less in lymphatic vessels with F8 and UEL than with comparable arteries or veins. ACE staining intensity, on the other hand, was similar in blood vessels and lymphatics. Both formalin and B5 fixation preserved antigenicity; however, background staining was greater with B5 fixation whereas tissue staining was slightly more intense with formalin fixation. PMID- 3037204 TI - Age-dependent changes in rat brain mitochondria of synaptic and non-synaptic origins. AB - Synaptic and non-synaptic mitochondria were isolated from the brains of 3- 12-and 28-30-month-old female Fisher 344 rats. Total oxygen consumption and oxygen consumption due to mitochondrial respiration decreases 83% with increasing age in synaptic mitochondria using malate plus glutamate as substrate, but only 33% in non-synaptic mitochondria; succinate-driven activity is not affected. Succinate driven superoxide generation decreases over 90% in both fractions; malate plus glutamate-driven superoxide generation decreases 50% in synaptic mitochondria only. The amount of c- and a-type cytochromes decreases approximately 50% in synaptic mitochondria. The absorbance wavelength maximum of cytochrome b decreases 2.6 nm in synaptic mitochondria from senescent brains but only 1.6 nm in non-synaptic mitochondria. PMID- 3037205 TI - Neuron-specific enolase (NSE) as a tumour marker and comparative evaluation with carcinoembryonic antigen (CEA) in small-cell lung cancer. AB - Serum neuron-specific enolase (NSE) was evaluated in a number of malignant tumours. It was elevated (greater than 12.5 micrograms l-1) in 13/17 (76.5%) patients with extensive small-cell lung carcinoma and in none of the three patients with limited disease. Of patients with carcinoma of the breast 4/12 (33.3%) had elevated concentrations. Normal concentrations were found in patients with non-Hodgkin's lymphoma (19) and Hodgkin's disease (15), carcinoma of the cervix (2), CSF and serum (5) of patients with gestational trophoblastic disease (with definite nervous system involvement). Comparative serial studies of NSE and carcinoembryonic antigen (CEA) concentrations were done in 15 patients with small cell lung cancer (SCLC). Of these 7/15 (46.7%) had elevated pre-treatment concentrations of both CEA and NSE, 1/15 (6.7%) had CEA elevated only, while 2/15 (13.3%) had NSE alone elevated. Of those patients with normal pre-treatment marker concentrations 3/5 (60%) had elevated markers on recurrence. The mean survival period was 61.9 weeks; 66.8 weeks for the marker-negative group and 44.6 weeks for the marker-positive (both NSE and CEA) group. Combined NSE and CEA evaluation provide additional means of monitoring SCLC. PMID- 3037207 TI - [Prevalence of opportunistic infections in risk groups for AIDS]. PMID- 3037208 TI - [Valproate therapy of ACTH hypersecretion]. PMID- 3037206 TI - Methotrexate for relapse of metastatic non-seminomatous germ-cell tumours. AB - Twelve patients with metastatic non-seminomatous germ-cell tumours who relapsed after 3-9 courses (median 7) of cis-platin containing combination chemotherapy, were treated with moderately high-dose methotrexate (1 g m-2) and folinic acid rescue. There was one partial response and the remaining eleven patients showed progression of disease. The patient that responded was one of two patients treated at the time of first relapse. It is concluded that methotrexate does not have a useful role in the salvage treatment of germ-cell tumours relapsing after cis-platin-containing chemotherapy or in patients who are primarily resistant to cis-platin-containing regimes. PMID- 3037209 TI - [Usefulness of gammagraphy with 201-thallium chloride and technetium 99m pertechnetate in the preoperative diagnostic localization of primary hyperthyroidism]. PMID- 3037211 TI - Age-associated pharmacodynamic changes. AB - Many pharmacodynamic changes with age have been identified. Quite likely, they, in concert with age-associated pharmacokinetic changes and multiple drug use, are responsible for altered drug action in the elderly and an increasing incidence of adverse drug reactions with age. The pharmacodynamic changes, which can result in altered action of important drugs, can lead to major adverse clinical outcomes if not recognized, particularly in those drugs which affect the heart and the central nervous system. PMID- 3037210 TI - [Cystic adenoid carcinoma. Apropos of 2 cases originating in the salivary glands]. AB - The CAQ is a tumor of the major and minor salivary glands, of superior airway glands, uterine cervix glands, and the Bartholin glands of the vulva. However, recently, primary cases of CAQ of the skin, originating in eccrine sweat glands were described. We report 2 CAQ cases, one, case 1, of minor salivary glands of the palate, that by extension, reached the skin of the face, finishing with kidney metastasis; and its histological type corresponded to the cribiform variety in both the primary tumor and the kidney metastasis. Case 2 presented in the parotid gland and its histological type is the solid variety of the CAQ. PMID- 3037212 TI - Gyrase-inhibitors impair caffeine elimination in man. PMID- 3037213 TI - [Heavy water (D20) inhibits growth of human xenotransplanted oropharyngeal cancers. An animal experiment study in nude mice]. AB - The large mass ratio of D to H, 2 to 1, gives rise to multiple effects in living organisms, when D is administered as heavy water (D2O). Deuterium at high concentration interferes with cell division and depresses the uptake of DNA precursors in mammalian cells (Katz et al., 1970). However, the effects of D2O on the organism disappear after cessation of deuteration of the body water. Mammals do not survive more than 35% of D2O substitution for normal water in their body fluids for long periods of time. However, mice drinking 30% D2O have a normal life span and normal body weight (Katz et al., 1970; Hodel et al., 1982). Heavy water also exerts antineoplastic effects. We demonstrated that moderate body deuteration combined with a cytostatic drug significantly increases the survival time of mice bearing transplantable murine neoplasms (Laissue et al., 1982). Hitherto, it is not known whether heavy water affects human neoplasms in the same way. Therefore, we have studied human squamous cell carcinomas of the oropharyngeal region which were xenografted in nude mice. A moderate deuteration of body water of the mice (less than or equal to 20 atom %) drinking 30% D2O, inhibited the tumour growth markedly by a factor of 0.4 to 0.5 compared to the growth of the same tumours in control animals drinking tap water. This effect of D2O seems to be directly proportional to the grade of malignancy of the carcinomas. Histologically, no differences were detectable between the original carcinomas and the tumour grafts in animals with and without D2O. PMID- 3037214 TI - Bone resorption in cholesteatoma: epithelial-mesenchymal cell interaction and collagenase production. AB - Clinical evidence has shown that chronic otitis media with cholesteatoma causes greater bone resorption than otitis media without cholesteatoma. What is the role of the epithelium and its products on the morbidity of cholesteatoma? We have studied the mechanism of collagenase production in cultures of rat epidermal cells and skin fibroblast-like cells under various conditions. The epithelial cells significantly induced mesenchymal cells to produce collagenase. We conclude that interaction between epithelial cells and fibroblast-like cells enhances production of collagenase. This enhancement of proteolytic activity may be crucial in the series of molecular events resulting in bone resorption associated with cholesteatoma. PMID- 3037215 TI - Regulation of rat adrenal medullary enkephalins by glucocorticoids. AB - Opioid peptides and their precursors of the proenkephalin family are found in the chromaffin cells of the rat adrenal medulla in low quantities. However, if the gland is denervated, there is a 10 to 20-fold increase in enkephalin-containing (EC) peptides consisting mostly of the precursor proenkephalin. The denervation induced rise in medullary EC peptides is blocked by hypophysectomy, and partially reinstated by corticosterone, dexamethasone or ACTH treatment. In the intact rat, intermediate doses of corticosterone or dexamethasone reduce the denervation induced increase in EC peptides, while a high dose of dexamethasone restores this response. These results indicate that glucocorticoids exert a permissive effect in vivo on the denervation-induced stimulation of EC peptide biosynthesis. PMID- 3037216 TI - Beta-adrenoceptors in human tracheal smooth muscle: characteristics of binding and relaxation. AB - Specific binding of [125I]-(-)-cyanopindolol to human tracheal smooth muscle membranes was saturable, stereo-selective and of high affinity (Kd = 5.3 +/- 0.9 pmol/l and RT = 78 +/- 7 fmol/g tissue). The beta 1-selective antagonists atenolol and LK 203-030 inhibited specific [125I]-(-)-cyanopindolol binding according to a one binding site model with low affinity in nearly all subjects, pointing to a homogeneous beta 2-adrenoceptor population. In one subject using LK 203-030 a small beta 1-adrenoceptor subpopulation could be demonstrated. The beta mimetics isoprenaline, fenoterol, salbutamol and terbutaline recognized high and low affinity agonist binding sites. Isoprenaline's pKH- and pKL-values for the high and low affinity sites were 8.0 +/- 0.2 and 5.9 +/- 0.3 respectively. In functional experiments isoprenaline relaxed tracheal smooth muscle strips having intrinsic tone with a pD2-value of 6.63 +/- 0.19. PMID- 3037217 TI - Inositol phospholipid hydrolysis in cultured astrocytes and oligodendrocytes. AB - Cultures of astrocytes and oligodendrocytes were prelabeled with 3H-inositol and the accumulation of 3H-inositol phosphates was determined following stimulation with a number of neuroactive substances. In astrocytes, norepinephrine (NE) produced the greatest stimulation with significant increase also observed with bradykinin. In oligodendrocytes, the greatest stimulation was produced by carbachol with significant increase also produced by bradykinin, histamine and NE. Carbachol was found to be ineffective in producing stimulation in astrocytes. The accumulation of 3H-inositol phosphates in astrocytes in response to NE was found to be dependent on the presence of Li+. The NE stimulation in astrocytes was dose-dependent and had an EC50 of 1.2 microM. This stimulation was blocked by the low concentration of the alpha 1-adrenergic antagonist prazosin but not by the alpha 2-adrenergic antagonist yohimbine. The NE-stimulated accumulation of 3H inositol phosphates in astrocytes was inhibited by the cyclic nucleotide phosphodiesterase inhibitor isobutylmethylxanthine as well as by the cAMP analog dibutyryl cAMP. PMID- 3037218 TI - Cardiovascular effects of delta 9- and delta 9(11)-tetrahydrocannabinol and their interaction with epinephrine. AB - The administration of delta-9-tetrahydrocannabinol (delta 9-THC, 0.078-5.0 mg/kg, i.v.) to rats anesthetized with pentobarbital caused as much as a 50% decrease in mean arterial blood pressure, heart rate and respiratory rate in a dose-dependent manner. Delta-9(11)-tetrahydrocannabinol (delta 9(11)-THC) was approximately 8 fold less potent than delta 9-THC in its hypotensive effect and had smaller effects on heart and respiratory rates that were not dose-related at doses below 5 mg/kg. Alternate injections of epinephrine (2 micrograms/kg) with vehicle and increasing cannabinoid doses (1.25-5.0 mg/kg) indicated a potentiation of both the duration of the pressor effect and the magnitude of the reflex bradycardic effect of epinephrine by both delta 9- and delta 9(11)-THC. Epinephrine also produced arrhythmias in rats receiving cannabinoids, but not in rats receiving alternate injections of vehicle. It is concluded that both cannabinoids have adverse effects on the cardiovascular system and adverse interactions with epinephrine in rats anesthetized with pentobarbital. PMID- 3037219 TI - [3H]adrenaline release from hypothalamic synaptosomes and its modulation by clonidine: effects of chronic antidepressant drug regimens. AB - [3H]Adrenaline ([3H]ADR, 40 nM) was accumulated by rat hypothalamic synaptosomes (P2) more rapidly and in significantly greater amounts than by similar preparations from cerebral cortex. There was no significant difference between these two tissues in the rate or amount of [3H]noradrenaline ([3H]NA, 40 nM) accumulation. Talusupram (10 microM), maximally inhibited the uptake of [3H]ADR into hypothalamic synaptosomes by 60%. Nomifensine further inhibited uptake by 14%. From these observations it was concluded that some [3H]ADR was accumulated into non adrenergic neuronal terminals. The effects of desipramine (DMI, 10 mg/kg/day and clorgyline (1 mg/kg/day) administration for 28 days on K+-evoked release of [3H]ADR was investigated using superfused hypothalamic synaptosomes. After both chronic antidepressant drug regimens, total [3H]ADR release (spontaneous + evoked) was significantly reduced. Evoked release of [3H]ADR (by KCl, 16 mM) was significantly reduced after the DMI but not the clorgyline regimens. Presynaptic alpha 2-adrenoceptor function in the hypothalamus was assessed during superfusion by measuring the reduction in K+-evoked release of [3H]ADR caused by clonidine (1 microM). The attenuating effects of clonidine on [3H]ADR release (42% in untreated controls and 36% after chronic clorgyline) was diminished (to 4%) after chronic DMI administration. Alpha 2 adrenoceptor numbers in the rat hypothalamus were not significantly changed after clorgyline or DMI administration, suggesting that the functional subsensitivity seen in synaptosomes after DMI, may not be related to alpha 2 adrenoceptor down regulation. PMID- 3037220 TI - Covalent labeling of opioid receptors with 3H-D-Ala2-Leu5-enkephalin chloromethyl ketone. I. Binding characteristics in rat brain membranes. AB - The chloromethyl ketone derivative of D-Ala2-Leu5-enkephalin was synthesized in a radioactive form, and the resulting compound (3H-DALECK) was used to label opioid receptors. 3H-DALECK binds with high affinity, specificity and saturability to rat brain membranes. The number of sites labeled is 130 fmoles/mg protein. Unlabeled opioids inhibited the binding of 3H-DALECK; etorphine and DAGO being most potent. A 10-fold preference for mu sites over delta was seen in site specific competition experiments; while DALECK displayed low affinity for kappa sites of rat brain. DALECK irreversibly blocked a certain population of sites. Approximately 40% of 3H-DALECK binding at 15 min, and 60% at 60 min association time did not dissociate in the presence of a large excess of unlabeled DALECK and was resistant to washing. Autoradiography performed after SDS-PAGE revealed specific alkylation of proteins with molecular weight of 74, 65, 56, 43 and 34 kD. These results demonstrate the applicability of using 3H-DALECK to covalently label opioid receptors. PMID- 3037222 TI - Identification and characterization of leukotriene C4 and D4 receptors on a cultured smooth muscle cell line, BC3H-1. AB - We studied the characteristics of the leukotriene (LT) C4 and D4 receptors on a cultured smooth muscle cell line, BC3H-1. Specific [3H]LTC4 binding to the cell membrane was greater than 80% of total binding and saturable at a density of 3.96 +/- 0.39 pmol/mg protein, with an apparent dissociation constant (Kd) of 14.3 +/- 2.0 nM (n = 9). The association and dissociation of [3H]LTC4 binding were rapid and apparent equilibrium conditions were established within 5 min. Calculated Kd value of [3H]LTC4 binding from the kinetic analysis was 9.9 nM. From the competition analysis, calculated Ki value of unlabeled LTC4 to compete for the specific binding of [3H]LTC4 was 9.2 nM and was in good agreement with the Kd value obtained from the Scatchard plots or kinetic analysis. The rank order of potency of the unlabeled competitors for competing specific [3H]LTC4 binding was LTC4 much greater than LTD4 greater than LTE4 greater than FPL-55712. The maximum number of binding sites (Bmax) of [3H]LTD4 in the membrane of BC3H-1 cell line was about 11 times lower than that of the [3H]LTC4. The calculated values of Kd and Bmax of [3H]LTD4 binding were 9.3 +/- 0.8 nM and 0.37 +/- 0.04 pmol/mg protein, respectively (n = 3). The rank order of potency or the unlabeled competitors for competing specific [3H]LTD4 binding was LTD4 = LTE4 greater than FPL-55712 much greater than LTC4. These findings demonstrate that BC3H-1 cell line possess both LTC4 and LTD4 receptors with a predominance of LTC4 receptors. Thus BC3H-1 cell line is a good model to study the regulation of LTC4 and LTD4 receptors. PMID- 3037221 TI - Covalent labeling of opioid receptors with 3H-D-Ala2-Leu5-enkephalin chloromethyl ketone. II. Binding characteristics in frog brain membranes. AB - 3H-D-Ala2-Leu5-enkephalin chloromethyl ketone (3H-DALECK) was used to label opioid receptors of frog brain membranes. We have previously shown (15) that 70% of the opioid receptors are of kappa type in this preparation. The binding of 3H DALECK was of high affinity, half maximal binding being achieved by 0.9 nM of the radioligand. The number of sites labeled was calculated to be 108 fmol/mg protein. Opioid ligands, incubated with the membranes prior to the label, inhibited 3H-DALECK binding with the following rank order:etorphine greater than EKC greater than DAGO greater than DALECK greater than DADLE. Dissociation experiments showed that 70% of the binding is irreversible. Fluorography performed after SDS-PAGE revealed specific covalent labeling of protein subunits of 90, 58 and 20 kD molecular weights. Results will be compared to those obtained in rat brain (13). Our two studies demonstrate that 3H-DALECK is a useful probe for investigation the subunit structure of opioid receptors. PMID- 3037223 TI - GABAA receptor-controlled 36Cl- influx in cultured rat cerebellar granule cells. AB - gamma-Aminobutyric acid (GABA)-mediated and bicuculline-sensitive 36Cl- influx and bicuculline-sensitive [3H] GABA binding were demonstrated in cultures of rat cerebellar granule cells. The addition of 10(-5) M GABA produced a two-fold increase in 36Cl-influx over the basal level and the maximal increase was observed after approximately 20 sec. Progressive occupation of GABAA receptor by [3H]-(1S-9R)-bicuculline methiodide decreased 36Cl- influx activated by 10 microM GABA. The above results suggest that primary cultures of rat cerebellar granule cells provide a new and reliable model for studying the GABA activated chloride fluxes. PMID- 3037224 TI - Effects of sodium on cell surface and intracellular 3H-naloxone binding sites. AB - The binding of the opiate antagonist 3H-naloxone was examined in rat whole brain homogenates and in crude subcellular fractions of these homogenates (nuclear, synaptosomal, and mitochondrial fractions) using buffers that approximated intra- (low sodium concentration) and extracellular (high sodium concentration) fluids. Saturation studies showed a two-fold decrease in the dissociation constant (Kd) in all subcellular fractions examined in extracellular buffer compared to intracellular buffer. In contrast, there was no significant effect of the buffers on the Bmax. Thus, 3H-naloxone did not distinguish between binding sites present on cell surface and intracellular tissues in these two buffers. These results show that the "sodium effect" of opiate antagonist binding is probably not a function of altered selection of intra- and extracellular binding sites. PMID- 3037225 TI - Temperature dependence and GABA modulation of [3H]triazolam binding in the rat brain. AB - The hypnotic triazolam (TZ), a triazolobenzodiazepine displays a short physiological half life and has been used for the treatment of insomnia related to anxiety states. Our major objectives were the direct measurement of the temperature dependence and the gamma-aminobutyric acid (GABA) effect of [3H]TZ binding in the rat brain. Saturation studies showed a shift to lower affinity with increasing temperatures (Kd = 0.27 +/- 08 nM at 0 degree C; Kd = 1.96 +/- 0.85 nM at 37 degrees C) while the Bmax values remained unchanged (1220 +/- 176 fmoles/mg protein at 0 degree C and 1160 +/- 383 fmoles/mg protein at 37 degrees C). Saturation studies of [3H]TZ binding in the presence or absence of GABA (100 microM) showed a GABA-shift. At 0 degrees C the Kd values were (Kd = 0.24 +/- 0.03 nM/-GABA; Kd = 0.16 +/- 0.04/+GABA) and at 37 degrees C the Kd values were (Kd = 1.84 +/- 0.44 nM/-GABA; Kd = 0.95 +/- 0.29 nM/+GABA). In contrast to reported literature, our findings show that TZ interacts with benzodiazepine receptors with a temperature dependence and GABA-shift consistent with predicted behavior for benzodiazepine agonists. PMID- 3037226 TI - gamma-Aminobutyric acid (GABA) mediated transmembrane chloride flux with membrane vesicles from rat brain measured by quench flow technique: kinetic homogeneity of ion flux and receptor desensitization. AB - Transmembrane chloride flux mediated by the GABAA receptor and the desensitization of the receptor were followed using quench flow technique with 36Cl- and a membrane preparation from rat cerebral cortex. Measurements in short times allowed these two processes to be resolved. In general the ion-flux activity was desensitized in two phases. A fast phase took place in circa 200 ms (100 microM GABA) followed by a slower phase in several seconds. A minority (10%) of the membrane preparations did not display the fast phase. It is desirable to be able to separate these two phases of desensitization to facilitate analysis of the responses of the receptor. A short preincubation with GABA removed the fast phase from a subsequent measurement. In the absence of the fast phase the whole ion-flux equilibration was seen as a single phase. The measurements presented covering a time range of 0.01 seconds to 10 seconds show a single phase of ion flux which can be described by a first order ion influx process and a single first order desensitization process with a half time of circa 1 s (100 microM GABA). The results imply a single kinetically homogeneous population of vesicles containing a single population of GABA receptor (remaining active) with a single phase of desensitization. An understanding of this homogeneity, and how to ensure it, gives a basis for quantitatively testing the effects of drugs on these responses. Ion flux measurements with quench flow technique are a suitable tool for investigation of the mechanism of action of neurotransmitter receptors from brain. PMID- 3037227 TI - Biphasic competition between opiates and enkephalins: does it indicate the existence of a common high affinity ("mu-1") binding site? AB - Displacement from brain membranes of labeled opiates by low concentrations of enkephalins and of labeled enkephalins by low concentrations of opiates has been previously explained by the existence of a common high affinity site termed mu-1. An alternative interpretation of the same results is that the trough seen in the low concentration zone of the displacement curves represents cross binding of mu and delta opioid ligands to delta and mu receptors, respectively. In three sets of experiments with brain membranes, the size of the trough is shown to be dependent on the labeled ligand used: The ratio between the size of troughs seen with [3H]D-Ala, D-Leu enkephalin and with [3H]morphine varies with experimental conditions (storage of membranes at 4 degrees C for 72 h), with ratio of mu:delta receptors (e.g. in thalamus and cortex which are enriched in mu and delta sites, respectively) and with pretreatment of membranes with naloxonazine. These results can not be explained by a common high affinity site, but rather by binding of [3H]D-Ala, D-Leu enkephalin to mu and of [3H]morphine to delta opioid receptors. PMID- 3037228 TI - Effect of central naloxone on hormone and blood pressure responses to hemorrhage in conscious sheep. AB - The role of the brain opioid system in the control of hypothalamic-pituitary adrenal activity was studied in 10 conscious sheep with an indwelling cannula in a cerebral lateral ventricle. On separate days, sheep received infusions of artificial CSF (control) and the opiate antagonist, naloxone (100 micrograms/hr) before and during acute moderate hemorrhage (15 ml/kg over 10 min). Infusion of naloxone before hemorrhage raised plasma ACTH and resulted in a significant increase in cortisol compared to the control infusion. In contrast, ACTH and cortisol responses to hemorrhage tended to be blunted by central naloxone infusion. The responses of vasopressin, aldosterone and the catecholamines remained unaffected by naloxone. The fall in blood pressure and the rise in heart rate accompanying hemorrhage were likewise unaltered. These results suggest that brain opioid peptides have an inhibitory effect on basal ACTH secretion but do not play a major role in modulating the hemodynamic or pituitary-adrenal responses to acute moderate hemorrhage in conscious sheep. PMID- 3037229 TI - 3'-5' cyclic-guanosine monophosphate increase in rat brain hippocampus after gamma-hydroxybutyrate administration. Prevention by valproate and naloxone. AB - An increase (123%) of cyclic GMP (cGMP) was observed in the hippocampus of the rat killed by microwave irradiation 45 min after administration of 500 mg/kg gamma-hydroxybutyrate (GHB) IP. This increase is time and dose dependent. No modification in cyclic nucleotide content was observed in striatum and in cerebellum. As the role of GHB has been implicated in neurotransmission, the fact that this compound increases cyclic GMP accumulation in hippocampus in vivo may represent a mechanism by which the actions of GHB are mediated at the cellular level. Valproate (400 mg/kg) or naloxone (10 mg/kg) pretreatment completely abolish the cGMP increase due to GHB. A GABAergic and/or opiate phenomenon may be involved in the mechanism of GHB induced increase of cGMP. PMID- 3037231 TI - Chronic administration of lithium modulates tryptophan transport by changing the properties of the synaptosomal plasma membrane. AB - The effect of chronic administration of lithium salts on the sodium dependent high-affinity system for tryptophan uptake was examined in plasma membrane vesicles derived from rat brain. Tryptophan transport was measured as a function of the temperature, Arrhenius plots of the data were prepared, and the apparent energies of activation were computed. Both plots were biphasic, the transition temperature decreasing from 23 degrees C in control animals to 18 degrees C in treated rats. The average apparent energies of activation for the carrier also change -both below and above the transition temperature- in treated animals when compared to controls. Our data support the idea that chronic administration of lithium induces a more fluid state of the synaptosomal plasma membrane that could explain many of the effects of Li+ on membrane-bound proteins. PMID- 3037230 TI - Modulation of phosphatidylinositol-4,5-bisphosphate hydrolysis in rat aorta by guanine nucleotides, calcium and magnesium. AB - Rat aortic smooth muscle homogenates and membrane preparations contain a phospholipase C which hydrolyzes phosphatidylinositol 4,5-biphosphate (PIP2). We discovered that guanyl-5'yl-imidodiphosphate (Gpp(NH)p) activated the hydrolysis of exogenous PIP2 but not of phosphatidylinositol (PI) in rat aortic membranes. Further, maximal Gpp(NH)p-dependent hydrolysis was dependent on physiological levels of calcium. Also, magnesium inhibited PIP2 hydrolysis and catalyzed the dephosphorylation of PIP2 to phosphatidylinositol-4-phosphate (PIP). The results imply that PIP2 is the primary substrate of the nucleotide-regulated phospholipase C in rat aorta and that calcium and magnesium are physiological regulators of this activity. PMID- 3037232 TI - Calmodulin antagonists suppress cholesterol synthesis by inhibiting sterol delta 24 reductase. AB - Preincubation of hepatoma cells and human skin fibroblasts in the presence of the calmodulin antagonists trifluoperazine and N-(6-aminohexyl)-5-chloro-1 naphthalene sulfonamide resulted in a dose-dependent suppression of [14C]mevalonolactone incorporation into cholesterol. At a calmodulin antagonist concentration of 25 mumol, the incorporation of [14C]mevalonolactone into cellular cholesterol was suppressed to about 30% (hepatoma cells) and 10% (human skin fibroblasts) of control values. When the total nonsaponifiable [14C]lipids were separated and analyzed by two-dimensional thin layer chromatography, an accumulation of [14C]desmosterol was observed along with reduced formation of [14C]cholesterol. However, when cells were preincubated in the presence of [14C]dihydrolanosterol, [14C]cholesterol formation was not inhibited by the calmodulin antagonists. About 25% of the cell-associated dihydrolanosterol radioactivity was converted to cholesterol in both control and calmodulin antagonist-pretreated cells. The data suggest that calmodulin antagonists prevent the conversion of desmosterol into cholesterol by inhibiting sterol delta 24 reductase and that the enzymes catalyzing sterol ring modifications are not affected by the inhibitors. PMID- 3037235 TI - Chemical-shift imaging with large magnetic field inhomogeneity. AB - A new echo-time-encoded chemical-shift imaging technique applicable to dual-peak spectroscopic imaging with large magnetic field inhomogeneity is proposed and studied. The basic concept and its applications to modest field homogeneity (approximately 3.0 ppm) as well as to relatively large field inhomogeneity (approximately 10.5 ppm) are discussed. Actual pulse sequences are given and some experimental results on human volunteers obtained with a 2.0-T KAIS NMR system are also presented. PMID- 3037233 TI - Periodate-induced lipid oxidation of erythrocyte membranes. AB - Exposure of human erythrocyte ghosts to 0.2-5 mM periodate at 0 C for 15 min resulted in an increase of thiobarbituric acid-reactive substances and fluorescent materials in the membrane. This increase was suppressed by radical scavengers, butylated hydroxytoluene and thiourea, indicating that periodate caused lipid oxidation of ghosts. A role of hemoglobin in the periodate-induced lipid oxidation of ghosts was suggested by the fact that the oxidation was augmented by hemoglobin and inhibited by an iron chelator, desferrioxamine. Treatment of ghosts with periodate caused membrane protein cross-linking with and without disulfide bridge. Erythrocytes were also susceptible to lipid oxidation by periodate, but only disulfide-mediated protein cross-linking was observed. Erythrocytes treated with neuraminidase or trypsin were less susceptible, but those treated with neuraminidase along with galactose oxidase were nearly as susceptible as untreated cells. The effect of galactose oxidase was diminished by reduction of the enzyme-treated cells with borohydride. These results indicate that the aldehyde moieties generated by periodate at the sialyl residues or those generated by galactose oxidase at the terminal galactosyl or N-acetyl galactosaminyl residues of the membrane glycoconjugates play a stimulating role in the periodate-induced membrane lipid oxidation. PMID- 3037234 TI - Comparison of the clearances of serum chylomicron triglycerides enriched with eicosapentaenoic acid or oleic acid. AB - Rat mesenteric lymph chylomicrons containing triglycerides enriched with either [14C]oleic acid (OA) or [14C]-eicosapentaenoic acid (EPA) were prepared by ultracentrifugation of lymph samples collected for 6 hr after a single duodenal infusion of an emulsion containing either fatty acid. These chylomicrons were injected into the jugular vein of recipient rats and, at various time intervals, blood was drawn and serum was assayed for radioactivity. In separate animals, serum lipoprotein fractions were separated by ultracentrifugation, and the redistribution of labeled fatty acid among circulating lipoproteins was determined by liquid scintillation spectrometry. When the early disappearance rates (10 min) of either total serum radioactivity or specifically the chylomicron fraction were compared, there were no differences between the groups receiving OA- or EPA-enriched chylomicrons. However, disappearance rates of EPA enriched chylomicrons were slower than those of OA-enriched chylomicrons from 25 to 90 min. The small but significant differences in the disappearance rates for the longer time periods cannot be ascertained without further studies. At 5 min after injection of either type of chylomicron, the d less than 1.006 g/ml lipoprotein fraction of serum chylomicrons and very low density lipoproteins contained almost 90% of the original radioactivity. By 240 min, when less than 2% of the radioactivity remained, this radioactivity in the d less than 1.006 g/ml fraction was 43-46%, with concomitant increases in the low and high density lipoprotein fractions and in the lipoprotein-free serum. PMID- 3037236 TI - Effect of glipizide on hepatic fructose 2,6-bisphosphate concentration and glucose metabolism. AB - Glipizide raised, in a dose-dependent manner, the concentration of fructose 2,6 bisphosphate in hepatocytes isolated from 24-hour fasted rats and incubated in the presence of 10 mmol/L glucose. Simultaneously, the rate of L-lactate production, as well as the rate of 3H2O formation from (3-3H)glucose, increased markedly. The concentration of glipizide calculated as corresponding to the half maximal effect in these metabolic parameters was 12 to 15 mumol/L. In hepatocytes isolated from fed rats, either normal or made diabetic by treatment with alloxan, glipizide inhibited the conversion of both (U-14C)pyruvate and (U-14C)lactate to (14C)glucose; an inverse correlation was established between hepatocyte fructose 2,6-bisphosphate levels and the rate of gluconeogenesis. The increase of fructose 2,6-bisphosphate concentration elicited by glipizide, which occurs without a significant modification of either 6-phospho-fructo 2-kinase activity or hepatocyte cyclic AMP levels, seems to be related to a significant accumulation of hexose 6-phosphates (glucose 6-phosphate and fructose 6-phosphate) in the hepatic cells. PMID- 3037238 TI - Immobilization of nucleoside diphosphatase at the allosteric site using pyridoxal 5'-phosphate derivatives. PMID- 3037237 TI - Impairments in hepatocyte phosphoinositide metabolism in endotoxemia. AB - The status of phospholipid metabolism and inositol lipids-mediated transmembrane signaling in rat hepatocytes was analyzed during chronic, nonlethal endotoxemia. Rats were infused intravenously (IV) with Escherichia coli endotoxin (ET) via subcutaneously implanted osmotic pumps at a rate of 0.1 mg/100 g bw/day. The experiments were performed after 30 hours of ET or sterile saline (NaCl) infusion, in hepatocytes prelabelled "in vitro" with 32P (15 microCi/mL) and further stimulated with vasopressin (VP, 0.23 mumol/L). Similar experiments were done with food-restricted animals, whose food intake was matched with the voluntary intake of ET-infused rats. Uptake of 32P label into phosphatidic acid (PA), phosphatidylinositol 4-phosphate (PIP), and phosphatidylinositol 4,5 bisphosphate (PIP2) occurs rapidly in cells from pair-fed, saline and ET-infused animals, and reaches a plateau between 60 and 80 minutes of incubation. Labeling of phosphatidylinositol (PI), phosphatidylethanolamine (PE), and phosphatidylcholine (PC) proceeds linearly after a ten-minute lag period for PI and 20 minutes for the two other lipids. The nutritional state greatly affects the distribution of 32P uptake into lipids, resulting in very low labeling of PA and PI and a high labeling of poly-PI as compared with control (taken from untreated rats) cells. In ET-v saline-infused rats, the labeling of PI and PE was depressed concomitantly with a proportional increase in the labeling of PIP and PC. The ability of VP to induce polyphosphoinositide (poly-PI) degradation in hepatocytes from saline-infused animals was similar to that observed in control cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3037239 TI - Tresyl chloride-activated supports for enzyme immobilization. PMID- 3037240 TI - Dextransucrase: a glucosyltransferase from Streptococcus sanguis. PMID- 3037241 TI - Measurement of lipid turnover in response to thyrotropin-releasing hormone. PMID- 3037242 TI - Hormone-regulated phosphoinositide turnover in permeabilized cells and membranes. PMID- 3037243 TI - Measurement of inositol phospholipid metabolites by one-dimensional thin-layer chromatography. PMID- 3037244 TI - Phosphatidylinositol kinases from rabbit reticulocytes. PMID- 3037245 TI - Hormonal activation of phosphoinositide breakdown in the presence of guanine nucleotides using a cell-free system from blowfly salivary glands. PMID- 3037246 TI - Guanine nucleotide regulatory proteins in receptor-mediated polyphosphoinositide hydrolysis in human leukocytes. PMID- 3037247 TI - Quantitative measurement of sn-1,2-diacylglycerols. PMID- 3037248 TI - Leukotriene B4 catabolism: quantitation of leukotriene B4 and its omega-oxidation products by reversed-phase high-performance liquid chromatography. AB - LTB4 and its omega-oxidation products may be rapidly, sensitively, and specifically quantitated by the methods of solid-phase extraction and reversed phase high-performance liquid chromatography (HPLC), which are described in this chapter. Although other techniques, such as radioimmunoassay or gas chromatography-mass spectrometry, may be utilized for quantitative analysis of the lipoxygenase products of arachidonic acid, only the technique of reversed phase HPLC can quantitate as many as 10 metabolites in a single analysis, without prior derivatization. In this chapter, we also reviewed the chromatographic theory which we utilized in order to optimize reversed-phase HPLC analysis of LTB4 and its omega-oxidation products. With this information and a gradient HPLC system, it is possible for any investigator to develop a powerful assay for the potent inflammatory mediator, LTB4, or for any other lipoxygenase product of arachidonic acid. PMID- 3037249 TI - Assay for the leukotriene B4 receptor. PMID- 3037251 TI - Measurement of phosphoinositide pools in WRK-1 cells. PMID- 3037250 TI - Measurement of key enzyme activities involved in the metabolism of platelet activating factor. PMID- 3037252 TI - Measurement of subcellular sites of polyphosphoinositide metabolism in isolated rat hepatocytes. PMID- 3037253 TI - Dihydropteridine reductase from bovine liver. PMID- 3037254 TI - 4-Hydroxyphenylpyruvate dioxygenase from human liver. PMID- 3037255 TI - 4-Hydroxyphenylpyruvate dioxygenase from Pseudomonas. PMID- 3037256 TI - 4-Hydroxyphenylpyruvate dioxygenase from avian liver. PMID- 3037257 TI - Monophenol monooxygenase from Neurospora crassa. PMID- 3037258 TI - Purification and properties of tryptophan side chain oxidase types I and II from Pseudomonas. PMID- 3037259 TI - Kynurenine-oxoglutarate aminotransferase from rat kidney. PMID- 3037260 TI - Aromatic-amino acid-glyoxylate aminotransferase from rat liver. PMID- 3037261 TI - Dehydroquinate synthase from Escherichia coli, and its substrate 3-deoxy-D arabino-heptulosonic acid 7-phosphate. PMID- 3037262 TI - 3-Dehydroquinate dehydratase from Escherichia coli. PMID- 3037263 TI - Shikimate kinases from Escherichia coli K12. PMID- 3037264 TI - Chorismate mutase-prephenate dehydratase from Escherichia coli. PMID- 3037266 TI - Prephenate dehydrogenase (monofunctional). PMID- 3037265 TI - Chorismate mutase from Streptomyces aureofaciens. PMID- 3037267 TI - Avian dopamine beta-monooxygenase. PMID- 3037269 TI - Aromatic amine dehydrogenase from Alcaligenes faecalis. PMID- 3037268 TI - Assay and purification of liver monoamine oxidase. PMID- 3037271 TI - Indoleamine N-methyltransferase from rabbit lung. PMID- 3037270 TI - Amine N-methyltransferases. PMID- 3037272 TI - Characterization of a pro-alpha 2(I) collagen gene mutation by nuclease S1 mapping. PMID- 3037273 TI - Proteoglycans of mineralized matrices. PMID- 3037274 TI - Transmembrane signaling in Dictyostelium. PMID- 3037275 TI - Identification of chemoattractant-elicited increases in protein phosphorylation. PMID- 3037276 TI - OmpR and EnvZ are pleiotropic regulatory proteins: positive regulation of the tripeptide permease (tppB) of Salmonella typhimurium. AB - The tppB locus of Salmonella typhimurium encodes the anaerobically-induced tripeptide permease. We have demonstrated that expression of tppB requires the function of the ompR and envZ gene products, originally identified as positive regulatory proteins required for the osmotic regulation of porin expression. Significantly, tppB expression is not osmotically regulated. We have also identified three additional genes whose expression depends on OmpR. Thus OmpR and EnvZ serve a more general regulatory role than has previously been supposed. This study provides the first detailed genetic analysis of the ompB locus of S. typhimurium. PMID- 3037277 TI - Unstable visible mutations induced in Drosophila melanogaster by injections of oncogenic virus DNA into the polar plasm of early embryos. AB - When RSV DNA cloned in pBR 322 or DNA of simian adenovirus Sa7 (C8) is injected into the pole plasm of embryos of various Drosophila stocks, the progeny of 1-70% of the surviving flies display visible mutations. The mutagenesis is partially directed: the loci mutating due to retrovirus and adenovirus DNA do not overlap. The majority of resulting mutants are characterised by high instability: reversions and new mutations occur in them, which sometimes spread over the whole population ("explosive" instability). The injected sequences are revealed by dot hybridization in the DNA of many mutant strains, but only rarely by Southern blotting procedures. The results show that the microinjection of oncovirus DNA into embryos is an approach for obtaining highly unstable strains even from wild type stable Drosophila stocks without crosses with MR lines or the introduction of P elements. The sets of unstable mutations induced by oncovirus DNA is different from those in hybrid dysgenesis. PMID- 3037279 TI - Sex steroid replacement in post-menopausal women: effects on thyroid hormone status. AB - We have measured serum thyroxine (T4), triidothyronine (T3), thyroid-stimulating hormone (TSH) free thyroxine (FT4) and thyroxine-binding globulin (TBG) levels in a total of 5 post-menopausal women who were receiving oestrogen alone (Premarin, n = 19), progestogen alone (Primolut-N, n = 12), a combination of oestrogen and progestogen (Prempak C, n = 14) or no treatment (control group, n = 12). No differences were observed between the Premarin and Prempak C groups; both exhibited elevated T4 and TBG levels, although free thyroxin (FT4) and T4/TBG concentrations were normal relative to those in the control group. The Primolut-N subjects showed subnormal T3 and FT4 levels relative to the controls. It was concluded that it is not possible to make general statements regarding the effects of sex steroids on FT4 levels. PMID- 3037278 TI - Transposon-induced symbiotic mutants of Bradyrhizobium japonicum: isolation of two gene regions essential for nodulation. AB - Two strains of the soybean endosymbiont Bradyrhizobium japonicum, USDA 110 and 61 A101 C, were mutagenized with transposon Tn5. After plant infection tests of a total of 6,926 kanamycin and streptomycin resistant transconjugants, 25 mutants were identified that are defective in nodule formation (Nod-) or nitrogen fixation (Fix-). Seven Nod- mutants were isolated from strain USDA110 and from strain 61 A101 C, 4 Nod- mutants and 14 Fix- mutants were identified. Subsequent auxotrophic tests on these symbiotically defective mutants identified 4 His- Nod- mutants of USDA110. Genomic Southern analysis of the 25 mutants revealed that each of them carried a single copy of Tn5 integrated in the genome. Three 61 A101 C Fix- mutants were found to have vector DNA co-integrated along with Tn5 in the genome. Two independent DNA regions flanking Tn5 were cloned from the three non auxotrophic Nod- mutants and one His-Nod- mutant of USDA110. Homogenotization of the cloned fragments into wild-type strain USDA110 and subsequent nodulation assay of the resulting homogenotes confirmed that the Tn5 insertion was responsible for the Nod- phenotype. Partial EcoR1 restriction enzyme maps around the Tn5 insertion sites were generated. Hybridization of these cloned regions to the previously cloned nod regions of R. meliloti and nif and nod regions of B. japonicum USDA110 showed no homology, suggesting that these regions represent new symbiotic clusters of B. japonicum. PMID- 3037280 TI - Follow CDC guidelines for protection from AIDS. PMID- 3037281 TI - The mode of entry of herpes simplex virus type 1 into Vero cells. AB - The mode of entry of herpes simplex virus type 1 (HSV-1) into Vero cells was investigated quantitatively with biological techniques. The entry of virus occurred rapidly when the virus-adsorbed cells were incubated at 37 C. The kinetics of virus entry was found to be similar to that of the process of uncoating, indicating that the uncoating of HSV-1 occurs simultaneously with the entry of virus into the cell. Experiments with ammonium chloride revealed that acidity in endosomes is not necessary for the entry or uncoating of HSV-1, in contrast with the cases of enveloped RNA viruses. In addition, endocytosis of the virus seems to be one of the processes of entry for HSV-1. However, the kinetics of endocytosis showed that the cell-bound virus is endocytosed gradually and suggested that the endocytosis of HSV-1 does not lead the virus to an uncoating process. These results are most consistent with a mechanism of entry for HSV-1 involving fusion of the viral envelope with the plasma membrane of the host cell. PMID- 3037283 TI - Total nucleotide sequences of the infectious cloned DNAs of bean golden mosaic virus. AB - Complete nucleotide sequences of the infectious cloned DNA components (DNA A and B) of bean bolden mosaic virus were determined. The DNA A (2585 nucleotides) and DNA B (2647 nucleotides) have little sequence homology with each other, but both A and B contain a common region of 205 nucleotides. A possible large hairpin structure is detected in the common region. Nucleotide sequences of DNAs A and B revealed the presence of 8 potential coding regions for proteins (m.w. greater than 10,000). Among them, four open reading frames (ORFs 1-4) encode proteins of m.w. 30,000 or greater, and are individually coded in virion DNA A and B senses (+) and their complementary senses (-), respectively. The other four ORFs 5-8 are in virion DNA B(+) and its complementary sense (-). All of the ORFs 1-4 have regulatory signals for RNA synthesis (TATAA/T) in the region 5' upstream from a potential start codon ATG. PMID- 3037282 TI - Effect of cytolytic infection on maintenance of resistance to HVJ (Sendai virus) in an altered BHK cell culture. AB - Altered baby hamster kidney (BHK-R) cells were serially cultured in the continuous presence of hemagglutinating virus of Japan (HVJ). These cells showed a distinct resistance to superinfection with the homologous HVJ. This resistance of BHK-R cells gradually disappeared after serial passages in the presence of ultraviolet-irradiated HVJ particles which lost infectivity but still preserved hemagglutinating and neuraminidase activities. When BHK-R cells were serially cultured in the presence of a temperature-sensitive mutant of HVJ at non permissive temperature, the cells also lost the resistance. The resistance of BHK R cells remained unchanged, even after prolonged incubation in virus-free maintenance medium under the conditions of no cell division. It was suggested that killing of virus-sensitive cells, which were generated during cell proliferation, was required for maintenance of the resistance. PMID- 3037284 TI - The role of ribosomes in the sensitivity of mycobacteria to tuberactinomycin. PMID- 3037285 TI - Papillomavirus--Dr Jekyll or Mr Hyde? PMID- 3037286 TI - Human papillomavirus infection of the cervix in Victoria, 1982-1985. AB - Cytological evidence of human papillomavirus infection was present in 4.1% of the smear-tests that were received by the Victorian Cytology (Gynaecological) Service during 1982-1985. The prevalence rate among women was estimated to be 2.6% in 1984; for one-third of the women this was a change from their previous cytological status. The prevalence of papillomavirus infection was noted to vary with age and referral source; higher proportions were seen in the age group, 20 24 years, and in the cytological results of women who were attending either a sexually-transmitted diseases clinic or Aboriginal health cooperatives. PMID- 3037287 TI - The clinical management and laboratory assessment of anal warts. AB - The results of a clinical and virological survey of anal warts from 85 predominantly homosexual or bisexual men is presented, together with an improved technique for the surgical treatment of these lesions. Two types of anal warts, which were classified as either "acute" or "chronic" on the basis of their appearance and clinical behaviour, were recognized commonly. However, our laboratory investigations--which consisted of the routine histopathological examination of all specimens, together with immunohistochemical testing for the common wart antigen of specimens from 30 patients and papillomavirus typing by human papillomavirus DNA probing of 27 specimens--failed to reveal any significant differences between the two classes of anal warts. By means of a dot hybridization technique with mixed human papillomavirus DNA probes for types 6 and 11 and types 16 and 18, all wart biopsy specimens that were tested were shown to contain human papillomavirus types 6/11; two specimens also contained human papillomavirus types 16/18. Southern blot hybridization studies of eight specimens revealed that five warts contained human papillomavirus type 6 only and three warts contained human papillomavirus type 11 only. Although the surgical technique that is described was successful in terms of patient acceptance and the eventual eradication of anal warts, there was a high rate of recurrence of the lesions, which necessitated repeat operations in two-thirds of the patients. The need for counselling before surgery and for regular follow-up examinations after surgery is discussed. PMID- 3037288 TI - Sensitivity in vitro of herpes simplex virus isolates to human fibroblast interferon. AB - Sensitivity of herpes simplex virus (HSV) isolates, from patients with recurrent infections, to human fibroblast beta interferon (IFN), was tested in vitro. Among 25 HSV strains, 12 were HSV-1, isolated from facial and labial lesions, and 13 were HSV-2 isolated from genital lesions. Viral sensitivity to IFN was examined on epithelial MB cell line by yield reduction and plaque reduction and close correspondence between the two methods was observed. Most of HSV-1 isolates were in the same range of sensitivity to IFN, while HSV-2 isolates varied in sensitivity and differences approached statistical significance (P = 0.14). No correlation was found between various biological properties, such as plaque size, virulence in baby mice, growth at different temperatures, thymidine kinase activity in the presence or absence of IFN and sensitivity to IFN. Application of beta IFN containing cream reduced clinical symptoms in most of the patients. However, larger numbers of patients should be evaluated in order to conclude whether in vitro sensitivity correlates to clinical improvement. PMID- 3037289 TI - Follow-up report on 50 subjects vaccinated against herpes genitalis with Skinner vaccine. AB - Fifty subjects at risk of herpes genitalis received 109 immunizations with Skinner herpes vaccine and were assessed after a follow-up period of 4-48 months, representing a total follow-up period of 694 patient months. There was no evidence of contraction of herpes genitalis in 49 subjects. The risk of virus transmission and rate of contraction of disease was quantified by construction of two functions, namely a unit of exposure risk calculated per year (UYE) and standard contraction rate (SCR); in this study the SCR was 0.02. There was no evidence of significant side-effects from vaccination. Administration of Alhydrogel adjuvant with vaccine induced temporary granuloma formation in most subjects but was only detectable beyond 1 year of follow-up in one subject, in whom a painless swelling of 0.2 cm was detected 3 years after vaccination. There was no evidence of immunological reactivity to host cell or calf serum antigens in any of the subjects vaccinated. PMID- 3037290 TI - [Long-term remission of metastasizing signet ring cancer in an unusual immune status]. PMID- 3037291 TI - Response of immunocompromised patients with acute herpes zoster to intravenous acyclovir. PMID- 3037292 TI - Vietnam nurses need to be welcomed home. PMID- 3037293 TI - Fighting sexism: an ongoing nurses' struggle. Part IV. PMID- 3037294 TI - [A case of necrosis of a hepatocellular carcinoma, caused by spasm of the celiac artery]. AB - A 54-year-old man was admitted to our hospital complaining of back pain and right hypochondrial pain. Ultrasonography and celiac angiography revealed a large tumor sized 9.4 X 8.1 cm. The tumor appeared hypervascular on angiogram. During the second angiography, an attempt at superselective hepatic angiography for the purpose of infusing a combination of Adriamycin and Lipiodol, spasm of the celiac artery occurred. High fever continued for 11 days after the spasm and serum transaminase was elevated. At the third angiography, the nature of the tumor was seen to have changed remarkably to one of hypovascularity. Percutaneous transhepatic tumor biopsy was done. Pathological diagnosis was necrosis of hepatocellular carcinoma. Due to heart disorders, ligation of the right hepatic artery was performed instead of hepatic resection. Postoperatively, the size of the tumor decreased further. It is thought that this patient had a tendency to suffer from vasospasm and that the tumor had a relatively low resistance to ischemia. PMID- 3037295 TI - [Intra-arterial chemoembolization with albumin microspheres including mitomycin C in inoperable hepatic cancer]. AB - Intra-arterial chemoembolization using mitomycin C microsphere (MMC MS) was carried out both for VX-2 tumor-bearing rabbits and for 19 patients with inoperable hepatic cancer. MMC MSs contain 5% MMC in albumin as a biodegradable drug carrier and an average diameter is 45 +/- 8 microns. VX-2 tumor, implanted into the hind legs of the rabbits, was treated intraarterially when it grew up to 2 cm in diameter. Tumor growth was compared with the other treated groups such as control, conventional MMC or blank microsphere; and MMC concentrations in the peripheral blood, femoral vein blood and muscles of the hind legs were measured after treatment. Tumor growth in the MMC MS group was remarkably retarded, but the other groups had no retardation. MMC concentrations of blood and muscle dropped below the assay limitation within 2 hours after conventional MMC injection, but in the animals treated with MMC MS, those showed the sustained drug release over for 6 hours. Fifteen patients with unresectable hepatic malignancies received intra-arterial hepatic infusion using conventional MMC, while 19 patients were treated by MMC MS infusion. Tumor regression in the 19 patients with MMC MS was seen in 42% (8/19), while that in the 15 with conventional infusion in 33% (5/15). Serum CEA levels in 12 patients with metastatic cancer decline from 57.7 +/- 72.2 ng/ml to 16.5 +/- 21.6 ng/ml 2 weeks after MMC MS treatment. However, those in 10 patients given conventional infusion dropped from 24.0 +/- 18.1 ng/ml to 17.4 +/- 18.0 ng/ml. The survival duration for patients given conventional infusion was 6.7 +/- 2.8 months, but that with MMC MS prolonged to 14.1 +/- 8.9 months. The MMC MS treatment for metastatic hepatic cancer had superiority to the conventional infusion treatment at p = 0.0040 in survival rate. PMID- 3037296 TI - Structural requirements for delta opioid receptor binding. AB - Structural features influencing opioid activity of enkephalin analogs were investigated through the synthesis and evaluation of opioid receptor binding affinities of a series of cyclic dithioether-containing analogs and structurally related linear analogs of the cyclic, disulfide-containing peptides, [D-Pen2, D Pen5]enkephalin and [D-Pen2, L-Pen5]enkephalin, where Pen (penicillamine) is beta, beta-dimethylcysteine. The major effect of increasing the ring size of the cyclic moiety from disulfide to dithioether analogs was a large decrease in delta opioid receptor binding affinity which suggests that relatively compact conformations of the peptide ligand are necessary for optimal binding to this receptor. The effect of bulky, hydrophobic residues at position 2 in the peptide chain was evaluated by preparing the linear analogs, [D-t-Leu2, D-t Leu5]enkephalin (t-Leu, 2-amino-3,3-dimethylbutanoic acid) and [D-Abu2, D-t Leu5]enkephalin (Abu, 2-aminobutanoic acid). The former analog was found to be 36 and 450-fold less potent at delta and mu receptor sites, respectively, than was the latter, suggesting that bulky side chain substituents in position 2 of enkephalin analogs lead to a deleterious steric interaction at delta and particularly at mu receptors. PMID- 3037297 TI - Comparative pharmacological properties and functional coupling of mu and delta opioid receptor sites in human neuroblastoma SH-SY5Y cells. AB - The characteristics of mu and delta opioid receptor sites present in human neuroblastoma SH-SY5Y cells were investigated using [D-Ala2-N-methyl-Phe4-Gly (01)5]enkephalin (DAGO) and [2-D-penicillamine, 5-D-penicillamine]enkephalin (DPDPE), which are the most selective radioligands available for mu and delta sites, respectively. Scatchard analysis of the saturation isotherms revealed high affinity binding to a single class of sites for both [3H]DAGO (mu) and [3H]DPDPE (delta). [3H]DAGO labeled twice the number of sites compared to the binding capacity of [3H]DPDPE, yielding a mu/delta ratio of 2:1. Selective suppression of [3H]diprenorphine binding by specific opioid "blocking" ligands also showed a predominance of mu receptors, representing 65-70% of the total opioid sites. Competition binding studies carried out with a series of opiates and opioid peptides displayed higher potencies of mu- and delta-selective ligands in displacing the specific binding of [3H]DAGO and [3H]DPDPE, respectively. The [3H]diprenorphine/agonist competition curves were biphasic, indicating the high and low affinity states of mu and delta receptor sites in SH-SY5Y cells. Guanine nucleotide and sodium had differential effects on the agonist affinity and the proportion of high affinity states of mu and delta receptors. The mu and delta receptor sites were shown to be functionally coupled to adenylate cyclase. All of these data support the independent existence of mu and delta receptor types in human neuroblastoma cells. SH-SY5Y cells, therefore, represent a suitable model for investigating opioid-mediated responses in nerve cell populations. PMID- 3037298 TI - Selective calmodulin inhibition toward myosin light chain kinase by a new cerebral circulation improver, Ro 22-4839. AB - Ro 22-4839, a new cerebral circulation improver, has shown to be a potent calmodulin antagonist toward myosin light chain kinase (MLCK). It inhibited in vitro activity of calmodulin-activated cyclic AMP phosphodiesterase isolated from either bovine heart or brain and ATP-induced superprecipitation of chicken gizzard actomyosin with respective IC50 values of 20 microM, 17 microM, and 2.0 microM. The inhibitory action of Ro 22-4839 on the contractile system of the smooth muscle was demonstrated directly by its inhibition of chicken gizzard MLCK. Ro 22-4839 was found to potently inhibit MLCK with an IC50 value of 3.1 microM but was unable to inhibit the activity of MLCK rendered Ca2+/calmodulin independent by limited tryptic digestion. The inhibition of MLCK induced by Ro 22 4839 was completely overcome by addition of excess calmodulin. In contrast, Ro 22 4839 hardly inhibited calmodulin-activated Ca2+, Mg2+-ATPase from rat erythrocyte membrane or adenylate cyclase from rat brain. Use of hydrophobic fluorescence probes showed that Ro 22-4839 binds to the hydrophobic region of calmodulin like other calmodulin antagonists, trifluoperazine and W-7. However, the precise binding site of Ro 22-4839 to calmodulin is different from those of trifluoperazine and W-7, as suggested from differing IC50 values of these compounds against the probes. We conclude that Ro 22-4839 inhibits calmodulin activated enzymes, most significantly of MLCK, highly specific to smooth muscle contractile systems by binding to the hydrophobic domain of the calmodulin and inducing its conformational change in the presence of calcium. PMID- 3037299 TI - Ligand effects on membrane lipids associated with sodium, potassium-activated adenosine triphosphatase: comparative spin probe studies with rat brain and heart enzyme preparations. AB - Physical properties of membrane lipids associated with rat or dog brain and heart Na+, K+-ATPase preparations were compared using an electron spin resonance probe, 5-doxyl stearate. The degree of acyl chain order of the membrane lipids was greater for brain enzyme than for heart enzyme preparations; membrane lipids in the rat heart enzyme preparations were the most disordered. In the absence of added ligands, membrane lipids did not appear to undergo a detectable temperature dependent rearrangement or structural transition. An apparent transition was observed in the simultaneous presence of Na+, Mg2+, and ATP. These ligands increased lipid order at temperatures above the structural transition, but not below it. In the presence of the above ligands, K+ caused a marked decrease in the transition temperature in the rat brain enzyme preparations, but only a modest decrease in rat heart enzyme preparations. Arrhenius plots of rat brain and heart Na+, K+-ATPase activity revealed a break point roughly corresponding to respective membrane lipid transition temperatures observed in the presence of Na+, K+, Mg2+, and ATP. A low concentration of ouabain (1 microM) failed to affect either the lipid transition temperature estimated by the spin probe or the value of lipid order of the rat brain enzyme preparations observed in the presence of Na+, Mg2+, and ATP, but markedly reduced the effect of K+ to lower the transition temperature observed in the presence of the above ligands. A high concentration (100 microM) of ouabain which was needed to completely inhibit rat heart enzyme eliminated the thermally induced structural rearrangement observed in the presence of Na+, Mg2+, and ATP, apparently through a nonspecific lipid perturbation. These results indicate that differences in the physical properties of the membrane lipids per se are unlikely to account for the low affinity of rat heart Na+, K+-ATPase for ouabain and also suggest that the use of high concentrations of ouabain required to completely inhibit Na+, K+-ATPase activity may cause nonspecific changes in addition to inhibition of Na+, K+-ATPase or the sodium pump. PMID- 3037300 TI - Treatment with dilute alkali-nuclease S1 permits the analysis of DNA damage: cells treated with platinum analogues. AB - We describe here an approach to characterize various lesions induced in DNA by drug treatments, using three parameters: (a) release of single-stranded DNA fragments by cell lysis in dilute alkali, which result from enzymatic strand scission during DNA repair or chemical alterations of DNA; (b) the presence of high molecular weight DNA in cells after lysis in dilute alkali followed by nuclease S1 treatment which, due to drug-induced DNA cross-links and its level is a measure of the amount of DNA-containing cross-links; and (c) the appearance of small double-stranded DNA fragments, when the cell lysis is followed by digestion with nuclease S1 to remove single-stranded DNA. This DNA shows the same characteristics as DNA of untreated cells, but it may contain monoadducts. By following the flow of label through the three parameters, one can characterize both the lesions induced in DNA and how the lesions are repaired. We report here results of three platinum analogues: cis-Pt(II), trans-Pt(II), and cis-FLAP(II). A large proportion of DNA in treated cells appears as fragments (parameter c). The cis- compounds and trans- compounds differ with regard to appearance of high molecular weight DNA (parameter b) and the initial release of fragments (parameter a). PMID- 3037301 TI - alpha-Naphthoflavone antagonism of 2,3,7,8-tetrachlorodibenzo-p-dioxin-induced murine lymphocyte ethoxyresorufin-O-deethylase activity and immunosuppression. AB - Suppression of murine humoral immunity by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) has been shown to occur in vivo and in vitro. Studies have indicated that suppression of humoral immunity is mediated by the Ah receptor. Data presented in this paper demonstrate that alpha-naphthoflavone (ANF) and beta-naphthoflavone (BNF), like TCDD, bind to rat and murine hepatic and murine splenocyte cytosolic Ah receptor. Furthermore, BNF induces cytochrome P1-450 monooxygenase activity as measured by ethoxyresorufin-O-deethylase (EROD) in murine spleen cells to the same extent as TCDD. In contrast, ANF predominantly acts to antagonize TCDD induction of splenocyte EROD activity. Examination of humoral immunity in vitro demonstrated that BNF, like TCDD, is suppressive. Whereas ANF is suppressive at cytotoxic concentrations, lower concentrations of ANF antagonize the suppressive effect of TCDD. Antagonism by ANF of TCDD-induced EROD activity and suppression of humoral immunity occur at similar concentrations. These data suggest that ANF blocks TCDD suppression of B lymphocyte differentiation by competing with TCDD for binding to the Ah receptor. Since the mechanism of TCDD toxicity is not fully understood, probes such as ANF may be of great use in examining the role of the Ah receptor in mediating toxicity. PMID- 3037302 TI - Characterization of a subline of P388 leukemia resistant to amsacrine: evidence of altered topoisomerase II function. AB - Sensitive (P388/S) and amsacrine-resistant (P388/amsacrine) sublines of P388 leukemia were cloned in vitro and tested for differential chemosensitivity against a panel of drugs. P388/amsacrine, resistant both in vivo and in vitro to amsacrine, was cross-resistant to other putative topoisomerase II inhibitors including teniposide, etoposide, bisantrene, and doxorubicin. P388/amsacrine, was however, as sensitive as cloned P388/S to camptothecin, an inhibitor of topoisomerase I. The pattern of cross-resistance suggested that an alteration in topoisomerase II may be involved in the resistance of P388/amsacrine to these drugs. No differences in the uptake of amsacrine were detected between the two sublines. Cross-resistance to vinblastine was evident in P388/amsacrine; however resistance to vinblastine was associated with alterations in uptake or efflux of the drug. The number of protein-concealed single-strand breaks induced in whole cells by amsacrine, teniposide, bisantrene, and camptothecin was measured. Diminished numbers of strand breaks in the resistant subline were consistent with decreases in DNA-protein crosslinks. In the absence of drug treatment, resistant cells sustained approximately one-half as many single-strand breaks and DNA protein crosslinks as the sensitive cells during preparation of nuclei. As measured by the P4 phage DNA unknotting assay, 0.35 M NaCl nuclear extracts from P388/S contained approximately 2.3-fold more topoisomerase II catalytic activity than did extracts from P388/amsacrine. The amount of protein that immunoreacted with a specific antibody to calf thymus topoisomerase II was also decreased in the resistant cells. These data suggest that alterations in topoisomerase II which lead to differential drug sensitivities are partially responsible for the resistance of P388/amsacrine to a specific group of drugs. PMID- 3037303 TI - Interactions of amiloride with alpha- and beta-adrenergic receptors: amiloride reveals an allosteric site on alpha 2-adrenergic receptors. AB - Interaction of amiloride with adrenergic receptors was studied using radioligand binding techniques. Amiloride competed for [3H]prazosin binding to alpha 1 adrenergic receptors on rat renal cortical membranes and BC3H-1 muscle cell membranes. Non-linear regression analysis of radioligand binding isotherms showed that amiloride increased Kd without a change in Bmax, suggesting the drug binds competitively in a mutually exclusive manner with the radioligand at the receptor binding site. Similarly, amiloride competitively blocked [125I]iodocyanopindolol binding to beta-adrenergic receptors on both tissues. The addition of guanylyl 5' imidodiphosphate or sodium chloride did not alter the interaction of amiloride with alpha 1- or beta-adrenergic receptors. The interaction of amiloride with alpha 2-adrenergic receptors was more complex and revealed an allosteric site. In both rat renal cortical membranes and intact human platelets, amiloride increased the Kd for [3H]rauwolscine binding, as well as decreasing the apparent Bmax. In binding experiments where amiloride competed for [3H]rauwolscine-binding sites, pseudo-Hill slopes of less than 1.0 were obtained for both platelet and renal alpha 2 receptors. In addition, amiloride increased the rate of [3H]rauwolscine dissociation from renal alpha 2 receptors. In the presence of 100-120 mM sodium chloride, the Ki for amiloride competition was decreased an average of 54% in renal membranes; in contrast, sodium increased the Kd of the agonist epinephrine. Taken together, these data support the hypothesis that alpha 2-adrenergic receptors, but not alpha 1- or beta-adrenergic receptors, have an allosteric site to which amiloride binds and which we propose to be a cation-binding site. PMID- 3037304 TI - Stereoselective formations of K-region and non-K-region epoxides in the metabolism of chrysene by rat liver microsomal cytochrome P-450 isozymes. AB - The K-region 5,6-epoxide and non-K-region 1,2- and 3,4-epoxides of chrysene were isolated by normal phase high performance liquid chromatography (HPLC) from a mixture of products formed in the metabolism of chrysene by liver microsomes from untreated (control), phenobarbital-treated, or 3-methylcholanthrene-treated rats in the presence of an epoxide hydrolase inhibitor, 3,3,3-trichloropropylene 1,2 oxide. Epoxides were characterized by ultraviolet, mass, and circular dichroism spectral and chiral stationary phase HPLC analyses. Each of the metabolically formed epoxides was hydrated by rat liver microsomal epoxide hydrolase to a trans dihydrodiol. The metabolically formed chrysene 5,6-epoxides were determined by chiral stationary phase HPLC and were found to contain (5S,6R):(5R,6S) enantiomer ratios of 68:32 (control), 71:29 (phenobarbital), and 5:95 (3 methylcholanthrene), respectively. The enantiomers of chrysene 1,2-epoxide and 3,4-epoxide were also resolved by chiral stationary phase HPLC. However, the enantiomeric compositions of the metabolically formed chrysene 1,2- and 3,4 epoxides, which racemized rapidly at room temperature, could not be directly determined. By using molecular oxygen-18 in the in vitro incubation of chrysene and by mass spectral analyses of the resulting oxygen-18-containing dihydrodiol metabolites and their acid-catalyzed dehydration (phenolic) products, both 1,2 epoxide and 3,4-epoxide were found to be converted by microsomal epoxide hydrolase-catalyzed water attack at predominantly (greater than or equal to 97%) the allylic carbons. PMID- 3037305 TI - Biochemical and morphological effects of sodium butyrate on Dictyostelium discoideum development. AB - Pretreatment of proliferating D. discoideum amoebae with 10 mM butyrate for at least 8 h (one duplicating time) induced a reversible and dose dependent premature expression of several developmental parameters when the cells were starved in the absence of the fatty acid. The aggregative phase of the morphogenetic cycle was reduced in 2 h and the appearance of mature fruiting bodies and spores took place 4 h earlier as a result of butyrate pretreatment. Some developmentally regulated proteins, such as contact-sites A, cell surface lectins and cyclic AMP phosphodiesterase were also expressed 2 h earlier in butyrate pretreated cells than in controls. The level of extracellular cyclic AMP was reduced in butyrate pretreated cells, while other parameters of cyclic AMP metabolism were not affected. Butyrate also caused a partial inhibition of growth and the hyperacetylation of histone H4 in growing amoeba. These results suggest that butyrate acts as an inducer of differentiation in D. discoideum and can therefore be used as an experimental tool in order to explore regulatory mechanisms operating in slime mold differentiation. PMID- 3037306 TI - Metabolic and secretory response of tumoral-insulin producing cells to D-fructose and D-galactose. AB - At variance with normal islet cells, tumoral insulin-producing cells of the RINm5F line were found to display a positive secretory response not solely to D glucose and D-mannose, but also to D-fructose and D-galactose. All hexoses increased the ATP/ADP ratio, exerted a sparing action upon the oxidation of endogenous nutrients in cells prelabelled with either L-[U-14C]glutamine or [U 14C]palmitate, increased the output of lactic acid and, as judged from data collected in the presence of D-[U-14C]hexoses, underwent oxidation in the RINm5F cells. The secretory response to these four hexoses appeared commensurate with the extent of their metabolic effects. PMID- 3037307 TI - Nucleosomal organization of a BPV minichromosome containing a human H4 histone gene. AB - To address the relationship between chromatin structure and histone gene expression, the nucleosomal organization of a cell cycle-dependent human H4 histone gene in a bovine papilloma virus (BPV) minichromosome was examined. The nucleosome repeat length of the human H4 histone gene, maintained as a stable episome in a C127 mouse cell line designated I-8, was compared with that of the chromosomal copy of the H4 gene in human (HeLa) cells. In both cell lines, the H4 histone gene is predominantly expressed during the S phase of the cell cycle. The nucleosome repeat length of total HeLa cell and C127 mouse cell chromatin was similarly examined. Nuclei were digested with micrococcal nuclease and the DNA was fractionated electrophoretically, transferred to nitrocellulose filters and hybridized with radiolabelled (32P) cloned DNA probes. The nucleosome repeat length of the H4 gene, as an episome in the C127 mouse cell (153 +/- 8) and as an integrated copy in a HeLa cell (163 +/- 10) was considerably shorter than total genomic host cell (C127) (190 +/- 5) or HeLa cell chromatin (183 +/- 7). Our results indicate that the episomal H4 histone gene is packaged as chromatin. Moreover, the shortened nucleosome repeat length of the H4 gene, both as an episome or integrated chromosome sequence, suggests that the repeat length is characteristic of the gene and may be functionally related to its cell cycle regulated expression. PMID- 3037308 TI - [Nucleotide sequence of a mutant allele and wild type allele SUP1 and comparison of transcripts of SUP1 and SUP2 genes]. AB - Nucleotide sequences of the yeast recessive suppressor gene SUP1 and one of its mutant alleles (supl-ts36) are compared. One open reading frame is found in the gene able to code 438 amino acid residues. The reading frame is not interrupted by introns. Mutant allele supl-ts36 contains one nucleotide change at position + 101 (T----C) inducing the exchange of leucine on serine in N-terminal part of the polypeptide product. The homology between the structure of SUP1 gene and two groups of proteins is found (1.tRNA-binding or nucleotide-binding proteins; 2. mitochondrial proteins coded by mitochondrial genome). The size of transcript for SUP1 gene is 1600 nucleotides corresponding to the coding region of the gene. For SUP2 gene two stable transcripts are found corresponding to approximately 2500 and approximately 1400 nucleotides. The homology between SUP1 and SUP2 genes is not found. The absence of splicing for both SUP1 and SUP2 genes transcripts is demonstrated in the experiments with RNA2 conditional mutants with impaired splicing. PMID- 3037309 TI - [Mechanism of reaction catalyzed by RNA-ligase from bacteriophage T4]. AB - The dissociation constants of the complexes of RNA-ligase with acceptors, donors and the adenylylated donor A(5')ppAp have been determined on the basis of the inhibition of ATP-pyrophosphate exchange reaction. The dissociation constants of the complexes of the enzyme with "poor" acceptors (oligouridilates) have been shown to be slightly different from those with "good" acceptors (oligoadenylates). The dependence of the reaction velocity of the formation of ligation products on the concentration of acceptors (pA)4, (pU)4 and the adenylylated donor A(5)ppAp has been studied. On the basis of the data obtained the conclusion about the random addition mechanism has been drawn. The reaction takes place in the steady-state conditions in the case of (pA)4 and in the equilibrium conditions--in the case of (pU)4. PMID- 3037310 TI - [1H- and 2H-NMR relaxation in serum albumin solutions]. AB - The proton and deuteron relaxation times T1 and T2 were investigated in water and heavy water solutions of fatless human serum albumin. The temperature, concentration and Larmor precession frequency dependences can be well described by the conception of fast exchange in a simple biphasic model of water molecules rotation in the first hydration layer with slight anisotropy of motion. In the protonated systems the intermolecular dipole-dipole relaxation mechanism must be taken into account. PMID- 3037311 TI - Saccharomyces cerevisiae mutants unresponsive to alpha-factor pheromone: alpha factor binding and extragenic suppression. AB - Mutations in six genes that eliminate responsiveness of Saccharomyces cerevisiae a cells to alpha-factor were examined by assaying the binding of radioactively labeled alpha-factor to determine whether their lack of responsiveness was due to the absence of alpha-factor receptors. The ste2 mutants, known to be defective in the structural gene for the receptor, were found to lack receptors when grown at the restrictive temperature; these mutations probably affect the assembly of active receptors. Mutations in STE12 known to block STE2 mRNA accumulation also resulted in an absence of receptors. Mutations in STE4, 5, 7, and 11 partially reduced the number of binding sites, but this reduction was not sufficient to explain the loss of responsiveness; the products of these genes appear to affect postreceptor steps of the response pathway. As a second method of distinguishing the roles of the various STE genes, we examined the sterile mutants for suppression. Mating of the ste2-3 mutant was apparently limited by its sensitivity to alpha-factor, as its sterility was suppressed by mutation sst2-1, which leads to enhanced alpha-factor sensitivity. Sterility resulting from each of four ste4 mutations was suppressed partially by mutation sst2-1 or by mutation bar1-1 when one of three other mutations (ros1-1, ros2-1, or ros3-1) was also present. Sterility of the ste5-3 mutant was suppressed by mutation ros1-1 but not by sst2-1. The ste7, 11, and 12 mutations were not suppressed by ros1 or sst2. Our working model is that STE genes control the response to alpha-factor at two distinct steps. Defects at one step (requiring the STE2 gene are suppressed (directly or indirectly) by mutation sst2-1, whereas defects at the other step (requiring the STE5 gene) are suppressed by the ros1-1 mutation. The ste4 mutants are defective for both steps. Mutation ros1-1 was found to be allelic to cdc39-1. Map positions for genes STE2, STE12, ROS3, and FUR1 were determined. PMID- 3037312 TI - Adeno-associated virus gene expression inhibits cellular transformation by heterologous genes. AB - In this paper we report that adeno-associated virus (AAV) genomes inhibit stable transformation by several dominant selectable marker genes upon cotransfection into mouse tissue culture cells. Cotransfection of AAV genomes also inhibited the expression of pSV2cat in transient assays. In both cases, the inhibitory effect was independent of AAV DNA replication but required the AAV p5 and p19 genes, which encode proteins required for AAV DNA replication and regulation of AAV gene expression. Finally, addition of a cloned E4 gene in the transfection experiments partially blocked the AAV-mediated inhibitory activities. PMID- 3037313 TI - Interruption of two immunoglobulin heavy-chain switch regions in murine plasmacytoma P3.26Bu4 by insertion of retroviruslike element ETn. AB - A number of moderately reiterated murine genetic elements have been shown to have structures like those of retroviral proviruses. These elements are thought to be transposons, although little evidence of their transposability exists. Two members of one of these families of reiterated elements, the ETn family, have inserted into separate immunoglobulin heavy-chain switch regions in the plasmacytoma P3.26Bu4. Switch regions are those DNA segments associated with each immunoglobulin heavy-chain gene in which the somatic recombinations that accompany the heavy-chain switch occur. This role in somatic recombination may be relevant to the ETn insertions into the switch regions in P3.26Bu4 DNA. P3.26Bu4 and a number of other B-lineage cells contain ETn transcripts. PMID- 3037314 TI - Cloning and characterization of BCY1, a locus encoding a regulatory subunit of the cyclic AMP-dependent protein kinase in Saccharomyces cerevisiae. AB - We have cloned a gene (BCY1) from the yeast Saccharomyces cerevisiae that encodes a regulatory subunit of the cyclic AMP-dependent protein kinase. The encoded protein has a structural organization similar to that of the RI and RII regulatory subunits of the mammalian cyclic AMP-dependent protein kinase. Strains of S. cerevisiae with disrupted BCY1 genes do not display a cyclic AMP-dependent protein kinase in vitro, fail to grow on many carbon sources, and are exquisitely sensitive to heat shock and starvation. PMID- 3037317 TI - Gene recombination in X-ray-sensitive hamster cells. AB - Recombination was measured in Chinese hamster ovary (CHO-K1) cells and in the X ray-sensitive mutants xrs1 and xrs7, which show a defect in DNA double-strand break repair. To assay recombination, pairs of derivatives of the plasmid pSV2gpt were constructed with nonoverlapping deletions in the gpt gene region and cotransferred into the different cell types. Recombination efficiencies, measured as the transformation frequency with a pair of deletion plasmids relative to that with the complete pSV2gpt plasmid, were about 6% in both CHO-K1 and the xrs mutants for plasmids linearized at a site outside the gpt gene. However, these efficiencies were substantially enhanced by the introduction of a double-strand break into the homologous region of the gpt gene in one of a pair of deletion plasmids before cotransfer. This enhancement was apparently only about half as great for the xrs cells as for CHO-K1, but variation in the data was considerable. A much larger difference between CHO-K1 and the xrs mutants was found when the DNA concentration dependence of transformation was explored. While the transformation frequency of CHO-K1 increased linearly with DNA concentration, no such increase occurred with the xrs mutants irrespective of whether complete plasmids or pairs of deletion plasmids were transferred. The fraction of cells taking up DNA, assayed autoradiographically, was similar in all cell types. Therefore we suggest that while homologous recombination of plasmid molecules may not be substantially reduced in the xrs mutants,processes involved in the stable integration of plasmid DNA into genomic DNA are significantly impaired. PMID- 3037315 TI - ret transforming gene encodes a fusion protein homologous to tyrosine kinases. AB - The ret transforming gene was activated by recombination between two unlinked segments of human DNA, most likely during transfection of NIH 3T3 cells. To further define this transforming gene, we isolated and sequenced ret cDNA clones. The nucleotide sequence indicates that the active ret transforming gene encodes a fusion protein with a carboxy-terminal domain which is 40 to 50% homologous to members of the tyrosine kinase gene family. This tyrosine kinase domain is preceded by a hydrophobic sequence characteristic of a transmembrane domain. Transcription of the ret tyrosine kinase sequence was detected in the SK-N-SH neuroblastoma, HL-60 promyelocytic leukemia, and THP-1 monocytic leukemia cell lines, but not in 25 other human tumor cell lines surveyed. The ret tyrosine kinase may thus represent a cell surface receptor which is expressed in a restricted range of human cells. PMID- 3037316 TI - Target sequences for cis-acting regulation within the dual promoter of the human c-myc gene. AB - Recombinant plasmids of the human c-myc promoter-leader region and the bacterial chloramphenicol acetyltransferase (cat) gene were constructed. After transfection into different rodent and human cells, the 862-base-pair (bp) PvuII fragment carrying both c-myc promoters and 350 bp of the untranslated leader conferred 1/15 to 1/30 of the CAT activity mediated by the simian virus 40 promoter. The presence of additional sequences upstream of the PvuII fragment had an overall negative effect on c-myc promoter activity detectable by titration analysis with small amounts of transfected plasmid DNA. The analysis of numerous deletion constructs in the c-myc promoter-leader region as well as S1 mapping experiments demonstrated that the high CAT activity depended largely on the presence of the second promoter. By cotransfection of c-myc-cat constructs with plasmids carrying different parts of the c-myc promoter locus, targets for positively acting cellular factors were identified. Two positive regulatory elements were mapped within the 862-bp PvuII fragment. One was localized within the 248-bp PvuII-SmaI fragment -101 to -349 bp upstream of the first cap site and the other within the 142-pb XhoI-NaeI fragment of the first exon, comprising positions -95 to +47 relative to the second cap site. We conclude that the dual promotor of the human c-myc gene represents a strong eucaryotic promotor regulated by cooperation of positively and negatively acting cellular transcription factors. PMID- 3037319 TI - DNA sequence analysis of spontaneous mutations at the aprt locus of hamster cells. AB - To determine the nature of spontaneous mutational events in cellular genes in hamster cells, mutant adenine phosphoribosyltransferase (aprt) genes were cloned and the regions to which we mapped alterations were sequenced. A variety of nucleotide changes were found to occur in the 12 mutant genes analyzed. Most mutations were simple base-pair substitutions-transitions (both G X C----A X T and A X T----G X C) and transversions. The only multiple mutation was a simple transition next to a single-base-pair insertion. Of the 12 mutations, 4 were more complex, involving small deletions or duplications. Two of these were similar to previously described deletions in that they occurred between short direct sequence repeats. No hot spots were detected. Three independent mutations were characterized at one restriction endonuclease site, although no other mutations were detected in the nucleotides surrounding this site in other mutant strains. At a functional level, sequence changes were either in exons (resulting in missense and, in one instance, nonsense mutations) or at splicing sites. PMID- 3037318 TI - Expression of the c-myc oncogene under control of an immunoglobulin enhancer in E mu-myc transgenic mice. AB - Transgenic mice bearing a cellular myc oncogene coupled to the immunoglobulin heavy-chain enhancer (E mu) exhibit perturbed B-lymphocyte development and succumb to B lymphoid tumors. To investigate how the enhancer has affected myc expression, we analyzed the structure and abundance of myc transcripts in tissues of prelymphomatous mice and in the lymphomas. Expression of the E mu-myc transgene appeared to be confined largely to B lymphoid cells, being dominant in bone marrow, spleen, and lymph nodes, with no detectable expression in T cells or other hematopoietic lineages examined. The myc transcripts initiated very predominantly at the normal myc promoters, although use of the more upstream myc promoter was accentuated and an enhancer-associated promoter may be used infrequently. The level of E mu-myc transcripts in the preneoplastic lymphoid tissues and in the E mu-myc tumors was not markedly higher than myc RNA levels in proliferating normal lymphocytes. Thus, enforced expression of structurally normal myc transcripts at only a modestly elevated level has profound biological consequences. The absence of detectable endogenous c-myc RNA in any tumor, or in preneoplastic bone marrow, supports a negative feedback model for normal c-myc regulation. PMID- 3037320 TI - Repression of quiescence-specific polypeptides in chicken heart mesenchymal cells transformed by Rous sarcoma virus. AB - Chicken heart mesenchymal cells do not proliferate in medium of physiological composition containing plasma (S. Balk, Proc. Natl. Acad. Sci. USA 77:6606-6610, 1980). To understand the molecular events involved in cell quiescence and in the initiation of cell division under physiological conditions, we examined the differences in the patterns of protein synthesis of quiescent, hormone stimulated, and Rous sarcoma virus-transformed chicken heart mesenchymal cells. We describe the expression of a 20,000-kilodalton (kDa) polypeptide actively synthesized by quiescent cells but not by their transformed counterparts. Normal chicken heart mesenchymal cells stimulated with epidermal growth factor and insulin also repressed the synthesis of the 20,000-kDa polypeptide while actively growing but synthesized increasing amounts of the protein at high cell density (confluence). The synthesis of the 20,000-kDa protein is not restricted to chicken heart mesenchymal cells, since confluent, density-arrested chicken embryo fibroblasts also expressed high levels of the protein. Transformed chicken heart mesenchymal cells and embryo fibroblasts did not synthesize the protein even at high cell density. The 20,000-kDa polypeptide accumulated in the culture medium. PMID- 3037321 TI - Ingi, a 5.2-kb dispersed sequence element from Trypanosoma brucei that carries half of a smaller mobile element at either end and has homology with mammalian LINEs. AB - A dispersed repetitive element named ingi, which is present in the genome of the protozoan parasite Trypanosoma brucei, is described. One complete 5.2-kilobase element and the ends of two others were sequenced. There were no direct or inverted terminal repeats. Rather, the ends consisted of two halves of a previously described 512-base-pair transposable element (G. Hasan, M.J. Turner, and J.S. Cordingley, Cell 37:333-341, 1984). Oligo(dA) tails and possible insertion site duplications suggested that ingi is a retroposon. The sequenced element appears to be a pseudogene copy of an original retroposon with one or more open reading frames occupying most of its length. Significant homologies of the encoded amino acid sequences with reverse transcriptases and mammalian long interpersed nuclear element sequences suggest a remote evolutionary origin for this kind of retroposon. PMID- 3037322 TI - Signal processing, glycosylation, and secretion of mutant hemagglutinins of a human influenza virus by Saccharomyces cerevisiae. AB - We investigated the nature of signal recognition, transport, and secretion of mutant hemagglutinins (HAs) of a human influenza virus by the yeast Saccharomyces cerevisiae. The cDNA sequences encoding variant forms of influenza HA were expressed in S. cerevisiae. The HA polypeptides (HA500 and HA325) that were synthesized with their N-terminal signal peptides were correctly targeted to the membrane compartment where they were glycosylated. In contrast, the HA polypeptides (HA484 and HA308) lacking the signal peptide were expressed in the cytoplasm and did not undergo any glycosidic modification, demonstrating the importance of the heterologous signal sequence in the early steps of translocation in S. cerevisiae. The analysis of the N-terminal amino acid sequence of HA500 and HA325 polypeptides demonstrated the correct cleavage of the signal peptide, indicating the structural compatibility of a heterologous signal peptide for efficient recognition and processing by the yeast translocation machinery. The membrane-sequestered and glycosylated HA polypeptides were relatively stable in S. cerevisiae compared with the signal-minus, nonglycosylated HA molecules. Although both the anchor-minus HA (HA500) and HA1 (HA325) polypeptides were targeted efficiently to the membrane, their glycosylation and transport patterns were shown to be different. During pulse chase, the HA500 remained cell-associated with no detectable secretion into the extracellular medium, whereas the HA325 secreted into the medium. Furthermore, only the cell-associated and secreted forms of HA325 and not HA500 appeared to have undergone hyperglycosylation with the extensive addition of high-molecular weight outer-chain mannans. Possible reasons for the observed phenotypic behavior of these two mutant HAs are discussed. PMID- 3037323 TI - Glucocorticoids enhance stability of human growth hormone mRNA. AB - We have studied the control of expression of the human growth hormone (hGH) gene introduced into the chromosomes of mouse fibroblasts. Cell lines transformed with the hGH gene expressed low levels of intact hGH mRNA and secreted hGH protein into the medium. Although the level of expression of hGH mRNA was low, the gene remained responsive to induction by glucocorticoid hormones. To localize the sequences responsible for induction and to determine the mechanism by which these cis-acting sequences enhance gene expression, we have constructed a series of fusion genes between the hGH gene and the herpes simplex virus (HSV) thymidine kinase (tk) gene. We have demonstrated that a fusion gene in which hGH cDNA is flanked at its 5' terminus by an HSV tk promoter and is flanked at its 3' terminus by 3' HSV tk DNA remains inducible by glucocorticoids. Our studies indicate that the hGH exons contain sequences which are responsible for glucocorticoid hormone induction. Pulse-chase experiments, in vitro nuclear transcription, and approach to steady-state measurements indicate that the mechanisms responsible for induction of the hGH cDNA fusion gene operate posttranscriptionally to enhance the stability of hGH mRNA. Moreover, this increased stability was associated with an increase in the length of the 3' poly(A) tail on hGH mRNA. PMID- 3037324 TI - Glucocorticoid-regulated compartmentalization of cell surface-associated glycoproteins in rat hepatoma cells: evidence for an independent response that requires receptor function and de novo RNA synthesis. AB - The role of glucocorticoid hormones in the compartmentalization of cell surface associated mouse mammary tumor virus (MMTV) glycoproteins was examined in M1.54, a cloned line of MMTV-infected rat hepatoma tissue culture cells. The expression of cellular [2-3H]mannose-labeled and cell surface 125I-labeled MMTV glycoproteins was examined throughout a time course of exposure to dexamethasone, a synthetic glucocorticoid. Posttranslational localization of cell surface MMTV glycoproteins required 6 h of exposure to hormone and occurred approximately 4 h after their initial production in an intracellular fraction. This regulated localization to the cell surface correlated with glucocorticoid receptor occupancy and was inhibited by exposure to RU 38486, a powerful antagonist of glucocorticoid-mediated responses. Cell surface immunoprecipitation demonstrated that actinomycin D, an inhibitor of de novo RNA synthesis, prevented regulated expression of cell surface viral glycoproteins, suggesting that newly synthesized cellular components mediate this process. The localization of cell surface MMTV glycoproteins appeared normal in a transcriptional variant (CR1) that produces basal levels of MMTV RNA and glycoprotein precursors in the presence of dexamethasone. Thus, regulated compartmentalization of viral glycoproteins is not an obligate consequence of a critical precursor concentration. Taken together, our results suggest that posttranslational trafficking of cell surface-destined MMTV glycoproteins resulted from an independent glucocorticoid hormone response that required receptor function and de novo RNA synthesis. PMID- 3037325 TI - Products of in vitro cleavage and polyadenylation of simian virus 40 late pre mRNAs. AB - Formation of mRNA 3' termini involves cleavage of an mRNA precursor and polyadenylation of the newly formed end. Cleavage of simian virus 40 late pre mRNA in a crude nuclear extract generated two RNAs, 5' and 3' half-molecules. These RNAs were unmodified and linear. The 5' half-molecule contained sequences upstream but not downstream of the poly(A) site and ended in a 3'-terminal hydroxyl. The 3' half-molecules comprised a family of RNAs, each of which contains only sequences downstream of the poly(A) site, and ends in a 5'-terminal phosphate. These RNAs differed only in the locations of their 5' terminus. The 3' terminus of the 5' half-molecule was the adenosine 10 nucleotides downstream of AAUAAA, at the +1 position. The 5' terminus of the longest 3' half-molecule was at +2. Thus, these two RNAs contain every nucleoside and phosphate of the precursor. The existence of these half-molecules demonstrates that endonucleolytic cleavage occurs near the poly(A) site. 5' half-molecules generated in the presence of EDTA (which blocks polyadenylation, but not cleavage) ended at the adenosine at position +1 of the precursor. When incubated in the extract under suitable conditions, they became polyadenylated. 5' half molecules formed in 3'-dATP-containing reactions contained a single 3' deoxyadenosine (cordycepin) residue added onto the +1 adenosine and were poor polyadenylation substrates. We infer that the +1 adenosine of the precursor becomes the first A of the poly(A) tract and provides a 3' hydroxyl group to which poly(A) is added posttranscriptionally. PMID- 3037327 TI - Restriction of P-element insertions at the Notch locus of Drosophila melanogaster. AB - P elements move about the Drosophila melanogaster genome in a nonrandom fashion, preferring some chromosomal targets for insertion over others (J. C. J. Eeken, F. H. Sobels, V. Hyland, and A. P. Schalet, Mutat. Res. 150:261-275, 1985; W. R. Engels, Annu. Rev. Genet. 17:315-344, 1983; M. D. Golubovsky, Y. N. Ivanov, and M. M. Green, Proc. Natl. Acad. Sci. USA 74:2973-2975, 1977; M. J. Simmons and J. K. Lim, Proc. Natl. Acad. Sci. USA 77:6042-6046, 1980). Some of this specificity may be due to recognition of a particular DNA sequence in the target DNA; derivatives of an 8-base-pair consensus sequence are occupied by these transposable elements at many different chromosomal locations (K. O'Hare and G. M. Rubin, Cell 34:25-36, 1983). An additional level of specificity of P-element insertions is described in this paper. Of 14 mutations induced in the complex locus Notch by hybrid dysgenesis, 13 involved P-element insertions at or near the transcription start site of the gene. This clustering was not seen in other transposable element-induced mutations of Notch. DNA sequences homologous to the previously described consensus target for P-element insertion are not preferentially located in this region of the locus. The choice of a chromosomal site for integration appears to be based on more subtle variations in chromosome structure that are probably associated with activation or expression of the target gene. PMID- 3037326 TI - Functional simian virus 40 T antigen is expressed in hybrid cells having finite proliferative potential. AB - Simian virus 40-transformed human cells fused with other independently derived simian virus 40-transformed cells and tumor-derived cells containing activated H ras and N-ras oncogenes yielded hybrids capable of indefinite division. Fusions with various other immortal cells yielded hybrids that had limited division potential. T antigen expressed in limited-division hybrids was functional for the induction of cellular DNA synthesis. PMID- 3037329 TI - Factor interactions at simian virus 40 GC-box promoter elements in intact nuclei. AB - Primer extension footprinting was used to probe late simian virus 40 regulatory elements in intact infected cell nuclei. Specific protection was observed over the viral "GC-box" transcription elements. The participation of the bound templates in gene activation is addressed by quantitation that shows that their abundance greatly exceeds that of transcription complexes but is comparable to that of open chromatin. PMID- 3037328 TI - The nontranscribed chicken calmodulin pseudogene cross-hybridizes with mRNA from the slow-muscle troponin C gene. AB - A chicken calmodulin pseudogene with no introns was previously shown to hybridize under stringent conditions with an mRNA species present in skeletal and cardiac muscles, yet it would not hybridize to calmodulin mRNA (J. P. Stein, R. P. Munjaal, L. Lagace', E. C. Lai, B. W. O'Malley, and A. R. Means, Proc. Natl. Acad. Sci. USA 80:6485-6489, 1983). Using the pseudogene as a probe, we isolated a full-length cDNA corresponding to this mRNA from a chicken breast muscle library and showed by sequence analysis that it encodes slow-muscle troponin C and not the pseudogene product. Hybridization between the calmodulin pseudogene and slow-muscle troponin C cDNA is due to a short region of high homology in those nucleotides that encode helices B and C of troponin C and calmodulin. Genomic Southern analysis showed the calmodulin pseudogene and the gene for slow muscle troponin C to exist as distinct single copies. PMID- 3037330 TI - Context-dependent gene expression: cis-acting negative effects of specific procaryotic plasmid sequences on eucaryotic genes. AB - A sequence element within pBR322 DNA mediates a cis-acting negative effect on expression from eucaryotic genes in transient expression assays. The negative element overlaps with sequences that inhibit DNA replication, but its effect is observed in the absence of detectable replication of transfected DNA. PMID- 3037332 TI - Nucleotides in the polyomavirus enhancer that control viral transcription and DNA replication. AB - The polyomavirus enhancer is required in cis for high-level expression of the viral early region and for replication of the viral genome. We introduced multiple mutations in the enhancer which reduced transcription and DNA replication. Polyomaviruses with these mutant enhancers formed very small plaques in whole mouse embryo cells. Revertants of the viral mutants were isolated and characterized. Reversion occurred by any of the following events: restoration of guanosines at nucleotide (nt) 5134 and nt 5140 within the adenovirus 5 E1A enhancer core AGGAAGTGACT; acquisition of an A----G mutation at nt 5258, which is the same mutation that enables polyomavirus to grow in embryonal carcinoma F9 cells; duplication of mutated sequences between nt 5146 and 5292 (including sequences homologous with immunoglobulin G, simian virus 40, and bovine papillomavirus enhancer elements). Reversion restored both the replicative and transcriptional functions of the viruses. Revertants that acquired the F9 mutation at nt 5258 grew at least 20-fold better than the original mutant in whole mouse embryo cells, but replicated only marginally better than the original mutant in 3T6 cells. Viruses with a reversion of the mutation at nt 5140 replicated equally well in both types of cells. Since individual nucleotides in the polyomavirus enhancer simultaneously altered DNA replication and transcription in specific cell types, it is likely that these processes rely upon a common element, such as an enhancer-binding protein. PMID- 3037331 TI - The v-fms oncogene induces factor-independent growth and transformation of the interleukin-3-dependent myeloid cell line FDC-P1. AB - The normal cellular counterpart of the v-fms oncogene product is a receptor for the mononuclear phagocyte colony-stimulating factor, CSF-1. An interleukin-3 (IL 3)-dependent mouse myeloid cell line, FDC-P1, was infected with a murine retrovirus vector containing v-fms linked to a gene encoding resistance to neomycin (neo). Infected cells selected for resistance to the aminoglycoside G418 contained few proviral DNA copies per haploid genome, expressed low levels of the v-fms-coded glycoprotein, remained IL-3 dependent for growth, and were nontumorigenic in nude mice. In contrast, infected cells selected for their ability to grow in the absence of IL-3 contained an increased number of proviral insertions, expressed high levels of the v-fms-coded glycoprotein, and were tumorigenic in nude mice. The IL-3-independent cells expressed IL-3 receptors of comparable number and affinity to those detected in uninfected FDC-P1 cells and did not produce a growth factor able to support replication of the parental cells. Thus, the synthesis of high levels of the v-fms gene product in FDC-P1 cells abrogated their requirement for IL-3 and rendered the cells tumorigenic by a nonautocrine mechanism. The data suggest that v-fms encodes a promiscuous tyrosine kinase able to transform cells of the myeloid lineage that do not normally express CSF-1 receptors. PMID- 3037333 TI - Molecular genetics of androgen-dependent and -independent expression of mouse sex limited protein. AB - Genes of the mouse S locus encoding C4 (the fourth component complement) and Slp (sex-limited protein) show extensive homology but are distinct in their function and regulation. In some mouse strains, such as B10.D2, Slp is androgen regulated, whereas in others, such as B10.W7R, expression of Slp is constitutive. We have previously shown that the B10.W7R strain has multiple Slp genes. In this report, we present the structure of the single C4 and four Slp genes of the B10.W7R S locus and compare the upstream flanking regions by partial sequence analysis and function in transfection assays. Of the four Slp genes, three (Slpw7.A, Slpw7.B, and Slpw7.C) have upstream and promoter regions very similar to those of C4. The fourth Slp gene (Slpw7.D) is instead virtually identical to the androgen regulated allele (Slpd from the B10.D2 mouse) in upstream regions. In particular, far-upstream sequences from both Slpd and Slpw7.D render the bacterial chloramphenicol acetyltransferase gene hormonally responsive upon transfection into mammary carcinoma cell lines. The upstream sequences between 2 to 3 kilobases of the Slp promoter initiate transcription from multiple sites when fused proximal to the chloramphenicol acetyltransferase gene, and these transcripts are threefold more abundant in the presence of androgen. This behavior is similar for Slpd and Slpw7.D, which suggests that Slpw7.D may be androgen regulated but that this is masked in vivo by constitutive expression of the other Slp genes. Nonhomologous recombination is implicated not only in expanding the copy number of C4 and Slp genes in the B10.W7R mouse but also in creating hybrid genes with regulatory features of C4 and structural features of Slp. PMID- 3037335 TI - B-cell nuclear proteins binding in vitro to the human immunoglobulin kappa enhancer: localization by exonuclease protection. AB - Proteins capable of interacting with the enhancer of the immunoglobulin kappa gene in vitro have been detected in extracts of nuclei from human B cells and from human, mouse, and rabbit spleens. The experiments, based on an exonuclease protection technique, demonstrate nuclear protein factors binding to a 30- to 35 base-pair domain containing both the simian virus 40 enhancer core element (TTTCCA) and the octamer CAGGTGGC that was previously identified as the consensus sequence for protein-binding sites in the murine immunoglobulin heavy-chain enhancer. This 30- to 35-base-pair domain in the human kappa enhancer is homologous to a site of protein binding detected in the murine kappa enhancer by other investigators using a gel retardation assay. Our results complement in vivo dimethyl sulfate footprinting studies of the human immunoglobulin kappa enhancer which demonstrated B cell-specific changes in guanine reactivity immediately 5' to the consensus octamer. Together, these findings suggest that DNA-binding proteins in B-cell nuclei interact with the 5' portion of the human kappa-gene enhancer. Such proteins could play a role in the B cell-specific transcription of the human immunoglobulin kappa gene. PMID- 3037334 TI - Structure and expression of the Saccharomyces cerevisiae CRY1 gene: a highly conserved ribosomal protein gene. AB - The Saccharomyces cerevisiae CRY1 gene encodes ribosomal protein rp59, a component of the 40S ribosomal subunit. Mutations in CRY1 can confer resistance to the alkaloid cryptopleurine, an inhibitor of the elongation step of translation. The nucleotide sequence of the cloned CRY1 gene was determined. The predicted amino acid sequence shows that CRY1 encodes a 14,561-dalton polypeptide that has 88% amino acid sequence homology to the hamster or human S14 ribosomal protein responsible for emetine resistance and 45% homology to Escherichia coli ribosomal protein S11. Analysis of the DNA sequences upstream from CRY1 revealed the presence of three sequences, HOMOL1 (consensus, A/TACATCC/TG/ATA/GCA), RPG (consensus, ACCCA/GTACATT/CT/A), and a thymine-rich sequence, found upstream of more than 20 other cloned yeast genes encoding components of the translational apparatus. We exploited the ability to assay the expression of CRY1 in vivo by using the cryptopleurine resistance phenotype to demonstrate that these three consensus sequences are necessary for the transcription of CRY1. We previously showed that the upstream promoter element of the yeast RP39A gene consists of these identical sequence motifs. Therefore, we suggest that these three sequences define a consensus promoter element for the genes encoding the yeast translational apparatus. CRY1 is one of several hundred yeast genes, including ribosomal protein genes, whose expression is transiently decreased 10-fold upon heat shock. We found that the HOMOL1 and RPG consensus sequences are not necessary for the heat shock response of CRY1. PMID- 3037337 TI - Conservation of promoter sequence but not complex intron splicing pattern in human and hamster genes for 3-hydroxy-3-methylglutaryl coenzyme A reductase. AB - Regulation of the expression of 3-hydroxy-3-methyglutaryl coenzyme A (HMG-CoA) reductase is a critical step in controlling cholesterol synthesis. Previous studies in cultured Chinese hamster ovary cells have shown that HMG-CoA reductase is transcribed from a cholesterol-regulated promoter to yield a heterogeneous collection of mRNAs with 5' untranslated regions of 68 to 670 nucleotides in length. Synthesis of these molecules is initiated at multiple sites, and multiple donor sites are used to excise an intron in the 5' untranslated region. In the current paper, I report that human HMG-CoA reductase gene resembles the Chinese hamster gene in having multiple sites of transcription initiation that are subject to suppression by cholesterol. The human gene differs from the hamster gene in that a single donor splice site is used to excise the intron in the 5' untranslated region. All of the resulting RNAs have short 5' untranslated regions of 68 to 100 nucleotides. This difference in the splicing pattern of the first intron is species specific and not a peculiarity of cultured cells in that HMG CoA reductase mRNAs from Syrian hamster livers resemble those of the cultured Chinese hamster ovary cells. Comparison of the DNA sequences of the HMG-CoA reductase promoters from three different species--humans, Syrian hamsters, and Chinese hamsters--shows a highly conserved region of 179 nucleotides that extends from 220 to 42 nucleotides upstream of the transcription initiation sites. This region is 88% identical between the human and Chinese hamster promoter. When fused to the coding region of the Escherichia coli chloramphenicol acetyltransferase gene, this highly conserved region of the reductase gene directs the cholesterol-regulated expression of chloramphenicol acetyltransferase in transfected hamster cells, further indicating the interspecies conservation of the regulatory elements. PMID- 3037336 TI - Multiple calmodulin mRNA species are derived from two distinct genes. AB - We have observed three calmodulin mRNA species in rat tissues. In order to know from how many expressed genes they are derived, we have investigated the genomic organization of calmodulin genes in the rat genome. From a rat brain cDNA library, we obtained two kinds of cDNAs (pRCM1 and pRCM3) encoding authentic calmodulin. DNA sequence analysis of these cDNA clones revealed substitutions of nucleotides at 73 positions of 450 nucleotides in the coding region, although the amino acid sequences of these calmodulins are exactly the same. DNA sequences in the 5' and 3' noncoding regions are quite different between these two cDNAs. From these results, we conclude that they are derived from two distinct bona fide calmodulin genes, CaMI (pRCM1) and CaMII (pRCM3). Total genomic Southern hybridization suggested four distinct calmodulin-related genes in the rat genome. By cloning and sequencing the calmodulin-related genes from rat genomic libraries, we demonstrated that the other two genes are processed pseudogenes generated from the CaMI (lambda SC9) and CaMII (lambda SC8) genes, respectively, through an mRNA-mediated process of insertions. Northern blotting showed that the CaMI gene is transcribed in liver, muscle, and brain in similar amounts, whereas the CaMII gene is transcribed mainly in brain. S1 nuclease mapping indicated that the CaMI gene produced two mRNA species (1.7 and 4 kilobases), whereas the CaMII gene expressed a single mRNA species (1.4 kilobases). PMID- 3037338 TI - Transcriptional regulation of an hsp70 heat shock gene in the yeast Saccharomyces cerevisiae. AB - The yeast Saccharomyces cerevisiae contains three heat-inducible hsp70 genes. We have characterized the promoter region of the hsp70 heat shock gene YG100, that also displays a basal level of expression. Deletion of the distal region of the promoter resulted in an 80% drop in the basal level of expression without affecting expression after heat shock. Progressive-deletion analysis suggested that sequences necessary for heat-inducible expression are more proximal, within 233 base pairs of the initiation region. The promoter region of YG100 contains multiple elements related to the Drosophila melanogaster heat shock element (HSE; CnnGAAnnT TCnnG). Deletion of a proximal promoter region containing one element, HSE2, eliminated most of the heat-inducible expression of YG100. The upstream activation site (UAS) of the yeast cytochrome c gene (CYC1) can be substituted by a single copy of HSE2 plus its adjoining nucleotides (UASHS). This hybrid promoter displayed a substantial level of expression before heat shock, and the level of expression was elevated eightfold by heat shock. YG100 sequences that flank UASHS inhibited basal expression of UASHS in the hybrid promoter but not its heat-inducible expression. This inhibition of basal UASHS activity suggests that negative regulation is involved in modulating expression of this yeast heat shock gene. PMID- 3037340 TI - Microinjection of ras p21 induces a rapid rise in intracellular pH. AB - Quiescent mouse NIH 3T3 cells responded to microinjection of activated ras p21 with a rapid and sustained rise in intracellular pH (approximately 0.17 pH units). The p21-induced pH change was inhibited by amiloride treatment or growth of cells in medium low in sodium, suggesting a role for the Na+/H+ antiporter. Amiloride was found to suppress p21-induced mitosis, also. PMID- 3037341 TI - Strontium phosphate transfection of human cells in primary culture: stable expression of the simian virus 40 large-T-antigen gene in primary human bronchial epithelial cells. AB - Strontium ion formed DNA-phosphate precipitates analogous to those formed by calcium but lacking the lethal and differentiation-inducing effects of calcium on many epithelial cell types in primary culture. Human primary bronchial epithelial cells were transiently and stably transfected by using strontium phosphate; the frequency of stable transformation with a plasmid carrying the simian virus 40 large-T-antigen gene was greater than 10(-4). PMID- 3037339 TI - Expression of three genes for elongation factor 1 alpha during morphogenesis of Mucor racemosus. AB - Three genes, TEF-1, -2, and -3, encode elongation factor 1 alpha in Mucor racemosus. Neutral and alkaline S1 nuclease analyses revealed that the genetic organization is unique for each of the genes. The number and size of the intervening sequences vary in these closely related genes, which suggests that complex genetic rearrangements gave rise to the elongation factor 1 alpha gene family. Nucleotide sequence data from restriction fragments isolated from the 5' and 3' ends of TEF-2 and -3 confirmed the presence of a second intervening sequence in these genes. These data along with S1 nuclease mapping revealed a region at the 3' end of the three genes which was predicted to be transcribed but untranslated. Unique oligonucleotides containing 19 bases were synthesized to hybridize to this unique trailer region in the elongation factor 1 alpha transcripts. These oligonucleotides were used as probes in standard Northern analysis of RNA purified from M. racemosus cells of several morphological types. It was determined that all three genes were expressed in the cell morphological types studied. However, the accumulated level of transcript derived from each gene varied considerably, with TEF-1 mRNA present in approximately twofold greater quantity than the TEF-3 transcript and up to sixfold greater quantity than TEF-2. The level of TEF-1 and -2 mRNA varied little among the cell morphological types studied, whereas TEF-3 mRNA was present in twofold greater quantity in sporangiospores than in either germlings or yeast cells which had been induced to undergo morphogenesis to hyphae. These data suggest that there is differential expression of the genes encoding elongation factor 1 alpha in M. racemosus. At least one gene, TEF-3, shows a morphology-specific pattern of transcript accumulation. PMID- 3037342 TI - Relatively stable population of c-myc RNA that lacks long poly(A). AB - We examined the turnover of c-myc RNA in the human promyelocytic cell line HL-60. In whole-cell RNA from rapidly growing cells we observed two major size classes of c-myc RNA, 2.4 and 2.2 kilobases (kb). When HL-60 cells were treated with actinomycin D for 30 min to inhibit transcription, the 2.4-kb c-myc RNA population was rapidly degraded, while the 2.2-kb c-myc RNA was degraded much more slowly. S1 nuclease transcript mapping and promoter-specific probes were utilized to show that both the stable 2.2-kb and the labile 2.4-kb c-myc RNA populations have 5' ends at the second promoter site (P2) and 3' ends at the second poly(A) addition site. To examine further possible structural differences between these two RNA populations, we fractionated RNA on an oligo(dT)-cellulose column to separate RNAs that contained long poly(A) tails from those which did not. We found that the labile 2.4-kb c-myc RNA population bound to oligo(dT) cellulose, while the more stable 2.2-kb c-myc RNA population did not. Preliminary estimates of their half-lives (t1/2) showed that the poly(A)+ c-myc RNA had a t1/2 of 12 min, while the c-myc RNA that did not bind to oligo(dT)-cellulose had a t1/2 of greater than 1 h. Several other cell types contain both poly(A)+ and nonpoly(A)+ c-myc RNAs including HeLa cells, normal human bone marrow cells, and normal mouse fetal liver cells. In agreement with the results in HL-60 cell, HeLa cell poly(A)+ c-myc RNA was more labile than c-myc RNA that lacked poly(A). The stable, nonpoly(A)+ c-myc RNA population may be important in the posttranscriptional regulation of c-myc expression. PMID- 3037343 TI - The highly conserved U small nuclear RNA 3'-end formation signal is quite tolerant to mutation. AB - Formation of the 3' end of U1 and U2 small nuclear RNA (snRNA) precursors is directed by a conserved sequence called the 3' box located 9 to 28 nucleotides downstream of all metazoan U1 to U4 snRNA genes sequenced so far. Deletion of part or all of the 3' box from human U1 and U2 genes drastically reduces 3'-end formation. To define the essential nucleotides within this box that direct 3'-end formation, we constructed a set of point mutations in the conserved residues of the human U1 3' box. The ability of the various mutations to direct 3'-end formation was tested by microinjection into Xenopus oocytes and transfection into HeLa cells. We found that the point mutations had diverse effects on 3'-end formation, ranging from no effect at all to severe inhibition; however, no single or double point mutation we tested completely eliminated 3'-end formation. We also showed that a rat U3 3' flank can effectively substitute for the human U1 3' flank, indicating that the 3' boxes of the different U snRNA genes are functionally equivalent. PMID- 3037344 TI - Functional expression of the cre-lox site-specific recombination system in the yeast Saccharomyces cerevisiae. AB - The procaryotic cre-lox site-specific recombination system of coliphage P1 was shown to function in an efficient manner in a eucaryote, the yeast Saccharomyces cerevisiae. The cre gene, which codes for a site-specific recombinase, was placed under control of the yeast GALI promoter. lox sites flanking the LEU2 gene were integrated into two different chromosomes in both orientations. Excisive recombination at the lox sites (as measured by loss of the LEU2 gene) was promoted efficiently and accurately by the Cre protein and was dependent upon induction by galactose. These results demonstrate that a procaryotic recombinase can enter a eucaryotic nucleus and, moreover, that the ability of the Cre recombinase to perform precise recombination events on the chromosomes of S. cerevisiae is unimpaired by chromatin structure. PMID- 3037345 TI - Structure of two developmentally regulated Dictyostelium discoideum ubiquitin genes. AB - A previously isolated cDNA clone, pLK229, that is specific for mRNA developmentally expressed during Dictyostelium discoideum spore germination and multicellular development, was used to screen two genomic libraries. Two genomic sequences homologous to pLK229 were isolated and sequenced. Genomic clone p229 is identical to the cDNA clone pLK229 and codes for a polypeptide of 381 amino acids. This polypeptide is composed of five tandem repeats of the same 76-amino acid sequence. Clone lambda 229 codes for a protein of 229 amino acids, containing three tandem repeats of the identical 76-amino-acid sequence. A computer search for homology to known proteins revealed that the 76-amino-acid repeat was identical to human and bovine ubiquitin except for two amino acid differences. PMID- 3037347 TI - Initiation of meiosis and sporulation in Saccharomyces cerevisiae does not require a decrease in cyclic AMP. AB - Meiosis and sporulation of Saccharomyces cerevisiae are initiated in a guanine auxotroph by guanine deprivation (E. Bautz Freese, Z. Olempska-Beer, A. Hartig, and E. Freese, Dev. Biol. 102:438-451, 1984). We used this condition to examine a hypothesis (K. Matsumoto, I. Uno, and T. Ishikawa, Cell 32:417-423, 1983) that initiation of meiosis requires a low level of cAMP. We found that, after guanine deprivation, the intracellular concentration of cAMP transiently decreased not more than 20% and not at all if the cAMP phosphodiesterase inhibitor 3-isobutyl-1 methylxanthine (IBMX) was added to the medium. Under these conditions, at least 76% of the cells sporulated in the absence of IBMX, and almost 100% sporulated in its presence. The sporulating cells continually excreted cAMP and utilized the gluconeogenic carbon source. The cells failed to sporulate efficiently and to form four-spored asci if simultaneously deprived of guanine and carbon. After guanine deprivation in glucose medium, sporulation remained suppressed and intracellular cAMP was unchanged. We conclude that, under conditions of guanine starvation, cAMP deficiency is not required for initiation of meiosis and sporulation, cAMP is produced in excess and excreted to the medium, the cells sporulate better if the cAMP concentration is increased by addition of IBMX, the cells require a gluconeogenic carbon source for complete and efficient sporulation, and suppression of sporulation by glucose is not mediated by cAMP. PMID- 3037346 TI - Drosophila melanogaster homologs of the raf oncogene. AB - A murine v-raf probe, representing the kinase domain, was used to identify two unique loci in Drosophila melanogaster DNA. The most closely related to v-raf was mapped by in situ hybridization to position 2F5-6 (Draf-1) on the X chromosome, whereas the other raf-related gene (Draf-2) was found at position 43A2-5 on chromosome 2. The nucleotide and amino acid homologies of Draf-1 to the kinase domain of v-raf are 61 and 65%, respectively. The large amount of a 3.2-kilobase Draf-1 transcript detected in eggs as a maternal message decreases during embryonic development, and significant steady-state levels are observed throughout the remainder of morphogenesis. We speculate that the Draf-1 locus plays an important role in early embryogenesis. PMID- 3037348 TI - Differential early viral gene expression in two stages of human papillomavirus type 16 DNA-induced malignant transformation. AB - Human papillomavirus (HPV) type 16 DNA induces progressive transformation in NIH 3T3 cells. Two types of cell lines, PM3T3G0 and PM3T3Fo, were isolated by G418 or focus selection, respectively, after transfection of cells by a recombinant HPV 16 DNA carrying the neo gene. These cell lines exhibited distinct phenotypes compared with controls. Saturation densities of PM3T3G0 and PM3T3Fo lines were two- to three- and five- to sevenfold greater than that of control NIH 3T3 cells, respectively. Neither cell type required high serum for growth, in contrast to NIH 3T3 cells. PM3T3G0 lines were premalignant, whereas PM3T3Fo lines manifested tumorigenicity within 2 weeks. Subpopulations of three PM3T3G0 lines underwent progressive transformation as reflected by focus formation. Analysis of HPV 16 specific mRNA species demonstrated that high levels of early and late gene expression were detected in premalignant PM3T3G0 lines, whereas relatively low quantities of selected gene messages were expressed in malignant transformants. Thus, high levels of viral gene expression are not crucial for malignant transformation. PMID- 3037349 TI - Expression of Rous sarcoma virus transforming protein pp60v-src in Saccharomyces cerevisiae cells. AB - The Rous sarcoma virus (RSV) pp60v-src protein was expressed in Saccharomyces cerevisiae cells either from a plasmid vector carrying the v-src gene or in yeast cells containing a single-copy v-src gene chromosomally integrated. In both yeast strains, v-src gene transcription is regulated by the galactose-inducible GAL10 promoter. Growth in galactose-containing medium resulted in constitutive expression of pp60v-src in the integrated strain and transient expression of higher levels of pp60v-src in the plasmid-bearing strain. The concentration of pp60v-src in the plasmid-bearing strain at its peak of expression was approximately threefold lower than that found in RSV-transformed mammalian cells. pp60v-src synthesized in yeast cells was phosphorylated in vivo on sites within the amino and carboxyl halves of the molecule. In immune complex kinase assays, the yeast pp60v-src was autophosphorylated on tyrosine and was able to phosphorylate exogenous substrates such as casein and enolase. The specific activity of pp60v-src synthesized in yeast cells was approximately 5- to 10-fold higher than that made in mammalian cells. Induction of pp60v-src caused the death of the plasmid-bearing yeast strain and transient inhibition of growth of the single-copy strain. Concomitantly, this induction resulted in high levels of tyrosine phosphorylation of yeast cell proteins. This indicates that pp60v-src functions as a tyrosine-specific phosphotransferase in yeast cells and suggests that hyperphosphorylation of yeast proteins is inimical to cell growth. PMID- 3037350 TI - Flounder antifreeze protein synthesis under heat shock control in transgenic Drosophila melanogaster. AB - The gene coding for the most abundant antifreeze protein (AFP) in the winter flounder was placed downstream of the Drosophila melanogaster hsp70 promoter and introduced into the D. melanogaster germ line by P-element-mediated transformation. In each of six transgenic strains tested, heat shock treatment induced the expression of two major AFP gene transcripts and one minor one. All three transcripts were spliced despite the lack of an obvious D. melanogaster internal intron-splicing sequence. The variation in transcript length was caused by selection of different polyadenylation sites. Western blots showed the presence of immunoreactive AFP in hemolymph from heat-shocked transformants. The immunoreactive material had a molecular weight of 6,200, which is consistent with the loss of the signal sequence from the primary translation product and the retention of the pro sequence. Thus, all the signals for flounder pre-mRNA and preprotein processing were recognized in D. melanogaster. PMID- 3037351 TI - Elements involved in oxygen regulation of the Saccharomyces cerevisiae CYC7 gene. AB - The CYC7 gene of Saccharomyces cerevisiae encodes the minor species, iso-2, of the cytochrome c protein. Its expression is governed by two regulatory sequences upstream from the gene: a positive site which stimulates transcription 240 base pairs 5' from the protein-coding sequence (-240) and a negative site which inhibits transcription at -300. In this study, the nature of the positive site and its relationship to the negative site has been investigated. Expression of the CYC7 gene is weakly inducible by oxygen. This effect was greatly enhanced by the semidominant CYP1-16 mutation in the trans-acting gene CYP1. The weak oxygen regulation in wild-type cells and the enhanced induction in CYP1-16 mutants were found to be mediated through the positive site. A mutational analysis of this site implicated at least part of a tandem, direct repeat of 9 base pairs as essential for the functioning of this site. The relationship between the positive and negative sites was investigated by comparing the expression of the intact gene with that of derivatives lacking either one or the other site. The expression of the gene containing only the negative site was actually stimulated anaerobically, while the gene containing the positive site alone, although having higher expression aerobically than anaerobically, had higher anaerobic expression than did the intact gene. Thus, it appeared that the combination of the positive and negative sites suppressed anaerobic expression. A model which attempts to explain these properties of the two sites and account for the regulation of the expression of the intact gene is presented. PMID- 3037353 TI - Cellular DNA rearrangements and early developmental arrest caused by DNA insertion in transgenic mouse embryos. AB - Insertional mutagenesis was investigated in a transgenic mouse strain (HUGH/4) derived from a fertilized egg injected with plasmid DNA containing the human growth hormone gene. Lethality occurred in homozygous embryos and was traced to the egg cylinder stage on days 4 to 5 of gestation, shortly after implantation. The mutation is on chromosome 12 and is distinct in location and integration pattern from another mutation also leading to lethality of homozygotes in the egg cylinder stage. Based on this and other evidence, relatively many genes may be recruited to activity near the time of implantation and may therefore present a large target of vulnerability to mutagenesis. The single insert in HUGH/4, consisting of approximately three tandem copies of plasmid sequences, is flanked by mouse cellular sequences that have undergone rearrangements, including a probable deletion. The data suggest the hypothesis that DNA rearrangements, which appear to be commonplace in transgenic mice, may arise because the initial insertional complex is unstable; stepwise changes may then be generated until a more stable conformation is achieved. PMID- 3037352 TI - Three Drosophila beta-tubulin sequences: a developmentally regulated isoform (beta 3), the testis-specific isoform (beta 2), and an assembly-defective mutation of the testis-specific isoform (B2t8) reveal both an ancient divergence in metazoan isotypes and structural constraints for beta-tubulin function. AB - The genomic DNA sequence and deduced amino acid sequence are presented for three Drosophila melanogaster beta-tubulins: a developmentally regulated isoform beta 3 tubulin, the wild-type testis-specific isoform beta 2-tubulin, and an ethyl methanesulfonate-induced assembly-defective mutation of the testis isoform, B2t8. The testis-specific beta 2-tubulin is highly homologous to the major vertebrate beta-tubulins, but beta 3-tubulin is considerably diverged. Comparison of the amino acid sequences of the two Drosophila isoforms to those of other beta tubulins indicates that these two proteins are representative of an ancient sequence divergence event which at least preceded the split between lines leading to vertebrates and invertebrates. The intron/exon structures of the genes for beta 2- and beta 3-tubulin are not the same. The structure of the gene for the variant beta 3-tubulin isoform, but not that of the testis-specific beta 2 tubulin gene, is similar to that of vertebrate beta-tubulins. The mutation B2t8 in the gene for the testis-specific beta 2-tubulin defines a single amino acid residue required for normal assembly function of beta-tubulin. The sequence of the B2t8 gene is identical to that of the wild-type gene except for a single nucleotide change resulting in the substitution of lysine for glutamic acid at residue 288. This position falls at the junction between two major structural domains of the beta-tubulin molecule. Although this hinge region is relatively variable in sequence among different beta-tubulins, the residue corresponding to glu 288 of Drosophila beta 2-tubulin is highly conserved as an acidic amino acid not only in all other beta-tubulins but in alpha-tubulins as well. PMID- 3037354 TI - Intermolecular recombination assay for mammalian cells that produces recombinants carrying both homologous and nonhomologous junctions. AB - We present an intermolecular recombination assay for mammalian cells that does not involve the reconstitution of a selectable marker. It is based on the generation of a shuttle vector by recombination between a bacterial and a mammalian vector. The recombinants can thus be amplified in mammalian cells, isolated by plasmid rescue in an Escherichia coli RecA- host, and identified by in situ hybridization, by using mammalian vector sequences as probes. Since both parental molecules can share defined lengths of homology, this assay permits a direct comparison between homologous and nonhomologous intermolecular recombination. Our results indicate that the dominant intermolecular recombination mechanism is a nonhomologous one. The relative frequency of homologous to nonhomologous recombination was influenced by the length of shared homology between parental molecules and the replicative state of the parental molecules, but not by the introduction of double-strand breaks per se. Finally, almost all of the recombinants with a homologous junction did not have the reciprocal homologous junction but instead had a nonhomologous one. We propose a model to account for the generation of these recombinants. PMID- 3037355 TI - 12-O-tetradecanoyl-phorbol-13-acetate induction of the human collagenase gene is mediated by an inducible enhancer element located in the 5'-flanking region. AB - Genomic clones coding for human fibroblast collagenase were isolated. By constructing and transfecting mutants with 5' and 3' deletion mutations of the 5' control region of the gene into human or murine cells, we delimited a 32-base pair sequence at positions -73 to -42 which is required for the induction of transcription by the tumor promoter 12-O-tetradecanoyl-phorbol-13-acetate. The DNA element behaves as a 12-O-tetradecanoyl-phorbol-13-acetate-inducible enhancer: it mediates the stimulation of transcription to the heterologous herpes simplex virus thymidine kinase promoter and acts in a position- and orientation independent manner. Differences in enhancer efficiency in different cell lines are interpreted to indicate differences in the activity of a trans-acting factor. PMID- 3037356 TI - [Decrease of plasma prolactin in valproate therapy--expression of an increase in the activity of the central GABAergic system]. AB - The effect of Valproat on plasma prolactin level in epileptic children with long term medication was investigated. Valproat was able to decrease plasma prolactin level significantly. This result is a sign of an increase in the central GABAergic system activity. PMID- 3037357 TI - Glomus tumor of the stomach. PMID- 3037358 TI - [Purification and substrate specificity of BcmI restriction endonuclease]. AB - A strain producing the new specific restriction endonuclease BcmI has been found in the Bacillus generum. The enzyme has been purified by chromatography on the blue sepharose, phosphocellulose PII, heparin sepharose. The analogous purification has been obtained when the blue sepharose has been substituted for the orange sepharose, the home produced sorbent. The BcmI enzyme has been shown by the substrate specificity definition to be an isoschizomer of the restriction endonuclease ClaI. PMID- 3037359 TI - [Construction of recombinant plasmids encoding the biosynthesis of the beta subunit of cholera toxin]. AB - The structure of the cholera toxin operon and the location of A and B toxin subunits have been studied by the Southern blot hybridization on filters. The gene coding for the synthesis of the cholera toxin B-subunit has been cloned in the vector plasmid pBR322. The structural gene of A-subunit has been partially deleted by the restriction endonuclease Bal31 digestion. The size of the 250 b. p. deletion has been defined by electron microscopy. The production of the cholera toxin B-subunit in Escherichia coli K12 cells has been studied. PMID- 3037360 TI - [Restriction analysis of the genome of swine abortion parvovirus and cloning of its PstI-EcoRI fragment]. AB - The cultured pig kidney cells infected by the porcine parvovirus (PPV) produced the virions and viral DNA. The latter was used as a matrix to synthesize the double stranded DNA. The obtained preparation is more homogenic than the natural replicative form and was used for restriction analysis of porcine parvovirus genome and for molecular cloning of its fragment. The isolated recombinant plasmids contained the PstI-EcoRI fragment of PPV DNA, containing 70% of the viral genome. The restriction analysis of replicative PPV DNA isolated from the infected cells has resulted in finding of the replicative form containing a 300 bp deletion in the 5'-region of PPV genome. PMID- 3037361 TI - [Directed mutagenesis of chromosomal genes of Escherichia coli in vivo: construction of RecA- and HtpR- mutants]. AB - The deletions in Escherichia coli chromosomal genes recA and htpR were constructed using the site-directed mutagenesis techniques. The obtained RecA- mutants are UV-sensitive and have a phenotype defective for the homologous DNA recombination. HtpR- mutant is temperature sensitive for growth and deficient in intracellular proteolysis. As a result a HtpR- mutant seems to be a preferable candidate for attempting to synthesize efficiently any alien protein in Escherichia coli cells. PMID- 3037362 TI - Test of models for the sequence specificity of UV-induced mutations in mammalian cells. AB - We have used mathematical modeling and statistical analysis to examine the correlation between UV-induced DNA damage and resulting base-substitution mutations in mammalian cells. The frequency and site specificity of UV-induced photoproducts in the supF gene of the pZ189 shuttle vector plasmid were compared with the frequency and site specificity of base-substitution mutations induced upon passage of the UV-irradiated vector in monkey cells. The hypothesis that the observed mutational spectrum is due to a preferential insertion of adenosine opposite UV photoproducts in the DNA template was found to best explain the mutational data. Models in which it was postulated that only (6-4) photoproducts, and not cyclobutane dimers, are mutagenic, or that the relative frequency of photoproduct formation does not influence mutation frequencies, fit the data much less well. This analysis demonstrates that molecular mechanisms of mutagenesis in mammalian cells can be deduced from mutational data obtained with a shuttle vector system. PMID- 3037363 TI - Pyridine prevents the clastogenicity of benzene but not of benzo[a]pyrene or cyclophosphamide. AB - Pyridine has been shown to be a much more potent inhibitor than other solvents of the metabolism and therefore the clastogenicity of benzene. In this report, pyridine prevented benzene-derived micronucleus formation in the bone marrow of ICR Swiss mice at much lower levels than xylene did. Time-course experiments did not indicate any delay in the peak micronucleus response to benzene caused by either pyridine or xylene. Similar experiments using pyridine with benzo[a]pyrene and pyridine with cyclophosphamide indicated that the effect of pyridine was specific for benzene. Benzo[a]pyrene (150 mg/kg) was inhibited by pyridine only at levels of 100 mg/kg or more, compared to inhibition of benzene (440 or 880 mg/kg) by pyridine at levels of 5 mg/kg. Cyclophosphamide was not inhibited at any level, and micronucleus formation was increased at lower ratios of pyridine to cyclophosphamide. These results provide indirect conformation of the work by others indicating that benzene is activated in part by a cytochrome P450 isozyme different from those activating benzo[a] pyrene or cyclophosphamide. Since DBA/2 mice (AHH-non-inducible) are more sensitive to benzene than C57Bl/6 mice (AHH inducible), single and multiple treatments with benzene were compared by micronucleus response in these two strains. DBA mice were more responsive in all cases. Pretreatment with methylcholanthrene caused a greater response to benzene in DBA/2 mice, suggesting that the cytochrome P450 isozyme involved is one of the forms induced by methylcholanthrene independent of the high-affinity Ah receptor. It is hypothesized that more efficient activation of benzene by the unique cytochrome P450 isozyme, perhaps combined with relatively less conjugation, may result in a greater sensitivity of the bone marrow versus the liver, and of DBA/2 versus C57Bl/6 mice. PMID- 3037364 TI - Enhanced reactivation of ultraviolet-irradiated human cytomegalovirus in normal host cells. AB - Enhanced survival of UV-irradiated human cytomegalovirus (HCMV) is demonstrated in normal human cells exposed to UV light prior to infection. The UV fluence that gave rise to maximum UV reactivation falls in the range of 15 J/m2. A large number of temperature-sensitive HCMV mutants were found under the peak of reactivation. These results confirm the existence of inducible SOS functions in human cells. PMID- 3037365 TI - Ionizing and ultraviolet radiation-induced reversion of sequenced frameshift mutations in Escherichia coli: a new role for umuDC suggested by delayed photoreactivation. AB - The ultraviolet (UV) and gamma radiation-induced reversion of the trpA21, trpA9813, and trpE9777 sequenced-frameshift mutations were studied in Escherichia coli K-12 with or without the plasmid pKM101. Radiation induced the reversion of all 3 frameshifts, and pKM101 enhanced this reversion 10-50-fold. Factors influencing the differential radiation revertability of frameshifts are discussed. The two most revertable frameshifts, trpE9777 and trpA9813, were used as probes to understand the role of the umuDC genes in radiation-induced frameshift reversion. Unlike the UV radiation-induced reversion of base substitution mutations, the reversion of these frameshifts was not enhanced in a uvrA umuC strain by photoreactivation after a post-UV-irradiation incubation. The UmuDC proteins are suggested to have functions in the radiation induction of frameshifts that are more complex than are their functions in the induction of base substitutions. PMID- 3037366 TI - Frameshift mutagenesis by nitracrine analogues in wild-type, uvrB, polA and recA strains of Salmonella typhimurium, with and without plasmid pKM101. AB - The mutagenic potential of 9-[(3-dimethylaminopropyl)amino]-acridine and its 1-, 2-, 3- and 4-nitro derivatives was studied in several strains of Salmonella typhimurium carrying the frameshift marker hisC3076. The strains all carried deep rough (rfa) mutations, and were either wild-type with respect to DNA repair capacity or carried recA, uvrB, polA1 or polA3 (amber) mutations. Derivatives with and without plasmid pKM101 were also studied. The des-nitro compound resembled 9 aminoacridine and other simple intercalating compounds. Both toxicity and mutagenesis were apparently unaffected by the uvrB and recA mutations or by the presence of plasmid pKM101. However, mutagenicity was reduced by the polA1 mutation, and virtually eliminated by the polA3 mutation. The drug was substantially more toxic in the latter, slightly more toxic in the former, of these polA- strains. Plasmid pKM101 enhanced mutagenesis and protected from toxicity in both polA1- and polA3- strains, although it did not restore either of these parameters to the level in the wild-type strain. The 2-nitro compound was generally similar to the des-nitro compound, except that it was considerably more toxic and apparently non-mutagenic in the recA-bearing strain. By contrast, mutagenicity of the 3- and 4-nitro compounds was enhanced by the uvrB mutation and by the presence of the plasmid. These compounds were highly toxic but non mutagenic in the recA- strain, and showed some increased toxicity in polA1- and polA3- strains. The 1-nitro compound has been previously found to cross-link DNA. Unlike well-characterised cross-linkers such as mitomycin C it was highly mutagenic in the uvrB- strain, and this mutagenesis was enhanced by plasmid pKM101, but eliminated by the recA mutation. At high doses, where the drug was completely toxic towards uvrB- or recA-carrying strains, it became mutagenic in the DNA-repair-proficient strains. This 'high-dose' mutagenesis was enhanced by plasmid pKM101, but reduced by the polA1 mutation and almost eliminated by the polA3 mutation. Although there are several possible interpretations of these data, they are compatible with the suggestion that the lesion induced by high doses (but not by low doses) of nitracrine is a cross-link, but that this is not the major mutagenic lesion. PMID- 3037367 TI - Comparison of the rep-38 and mmrA1 mutations of Escherichia coli. AB - The rep-38 and mmrA1 mutations are located very close to each other (approximately 85 min), and have been suggested to be allelic. To address this question we have compared the phenotypes of the mmrA1 and rep-38 mutants. Both the mmrA1 and rep-38 mutations blocked the enhanced killing and inhibition of postreplication repair by rich growth medium that occurs in UV-irradiated Escherichia coli K-12 uvrA cells, i.e., the mmrA1 and rep-38 strains did not show minimal medium recovery (MMR). However, phi X174 bacteriophage propagated well in mmrA1 strains, but not in rep-38 strains; a rep mutation sensitized a uvrA strain to UV irradiation, but a mmrA mutation did not. During chloramphenicol treatment, the rep-38 strain showed a larger amount of residual DNA synthesis than observed in the mmrA1 strain. The mmrA1 mutation appears to be a dominant mutation. This was determined by the failure of either plasmid pLC44-7 or episome F'KLF11, both of which carry the mmrA+ gene, to complement the Mmr- phenotype of a uvrA mmrA strain. Plasmid pLC44-7 is known to complement the rep-38 mutation, suggesting that rep-38 is a recessive mutation. Although certain of the phenotypes of the rep and mmrA mutants are similar, a number are quite different. These differences suggest that these two mutations are not allelic. PMID- 3037368 TI - Partial purification and characterization of cAMP-dependent protein kinase from Fasciola hepatica. AB - Three cyclic AMP (cAMP)-dependent protein kinases, designated A1, A2, and B, were isolated from the liver fluke Fasciola hepatica using Phenyl-Sepharose and DEAE cellulose chromatography. These enzymes differed with respect to activation by cAMP and their molecular weights. The half-maximal activation constant for cAMP dependent protein kinases A1 and B was 20 nM, while that of A2 was about five fold higher (110 nM). The estimated molecular weights for cAMP-dependent protein kinases A1 and A2 (both 98,000) suggest a dimeric form for these enzymes; whereas, the higher molecular weight for cAMP-dependent protein kinase B (187,000) indicates that this enzyme is a tetramer. The physical and kinetic properties of the catalytic subunit of fluke cAMP-dependent protein kinase were similar to those reported for the mammalian enzyme. The molecular weight of the catalytic subunit was estimated to be 41,000. The pH optimum for the enzyme was 6.0, 6.5, or 7.0 when casein, histone, or protamine were used as substrates. The protein substrate specificity was in the order histone greater than arginine-rich histone greater than casein greater than protamine greater than lysine-rich histone. Free Mg2+ 'stimulated' enzyme activity at low concentrations (0.5 to 5 mM), whereas at higher concentrations (greater than 5 mM) it became inhibitory. Of the divalent cations tested, only Co2+ and Mn2+ could substitute for Mg2+. Kinetic studies indicated that the reaction mechanism of this enzyme is sequential and that MgATP and MgADP are competitive ligands. Reconstitution experiments using the subunits of fluke and bovine heart cAMP-dependent protein kinase showed that there is sufficient structural homology between these enzymes such that the catalytic subunit from one species can combine with the regulatory subunit of the other species to form inactive holoenzyme. Thus, the present results indicate that cAMP-dependent protein kinase from F. hepatica is similar but not identical to the mammalian enzyme. PMID- 3037369 TI - Phosphomonoesterases of Leishmania mexicana mexicana and other flagellates. AB - Amastigotes and log-phase promastigotes of Leishmania mexicana mexicana contained distinct acid phosphatase, 3'-nucleotidase and 5'-nucleotidase activities, distinguishable by their response to pH and inhibitors. Both tartrate-sensitive and tartrate-resistant acid phosphatase were present in the two forms, amastigotes possessed less tartrate-resistant acid phosphatase than promastigotes. A tartrate-sensitive acid phosphatase was secreted into the medium in large amounts during the growth in vitro of L. m. mexicana promastigotes. The 5'-nucleotidase activity of both parasite forms was inhibited by ammonium molybdate, sodium tartrate and, to less extent, by sodium fluoride whereas 3' nucleotidase was inhibited by EDTA. All three activities were shown to be present on the external surface of both amastigotes and promastigotes. The three phosphomonoesterase activities were also detected in extracts of L. m. amazonensis, L. donovani, L. tarentolae, Crithidia fasciculata, Herpetomonas muscarum muscarum, H.m. ingenoplastis and Trichomitus batrachorum whereas 5' nucleotidase was not detected in Trypanosoma brucei brucei extract and 3' nucleotidase was absent from extracts of Trichomonas vaginalis and Tritrichomonas foetus. PMID- 3037370 TI - Case records of the Massachusetts General Hospital. Clinicopathological exercises. Case 29-1987. A 26-year-old man with inflammatory bowel disease and obstructive jaundice. PMID- 3037371 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 30-1987. A 43-year-old woman with breast cancer and the abrupt onset of respiratory failure. PMID- 3037372 TI - Lack of tocopherol in peripheral nerves of vitamin E-deficient patients with peripheral neuropathy. AB - Vitamin E deficiency is often associated with symptoms of a peripheral neuropathy. To evaluate whether vitamin E deficiency affects the vitamin E content of the peripheral nervous system, we measured the alpha-tocopherol content in biopsy specimens of sural nerve and adipose tissue from 5 patients with symptomatic vitamin E deficiency (2 with homozygous hypobetalipoproteinemia and 3 with familial isolated vitamin E deficiency) and 34 control patients with neurologic diseases without vitamin E deficiency. A significant reduction in tissue tocopherol content was present in the vitamin E-deficient patients, as compared with the controls, both in sural nerves (1.8 +/- 1.2 vs. 20 +/- 16 ng per microgram of cholesterol [P less than 0.001], or 7.7 +/- 5.4 vs. 64 +/- 44 ng per milligram of wet weight [P less than 0.01]) and in adipose tissue (46 +/- 43 vs. 222 +/- 111 ng per milligram of triglyceride [P less than 0.001]). Levels of tocopherol in adipose tissue were significantly correlated (P less than 0.001) with levels in peripheral nerves. The low tocopherol content of the nerves preceded histologic degeneration in three vitamin E-deficient patients, suggesting that the nerve injury resulted from the low nerve tocopherol content. PMID- 3037373 TI - Chronic bone marrow failure due to persistent B19 parvovirus infection. PMID- 3037374 TI - Effects of adrenergic and cholinergic agents and leukotrienes on mucociliary transport force measured by using frog palate. PMID- 3037375 TI - Ia binding ligands and cAMP stimulate nuclear translocation of PKC in B lymphocytes. AB - Altered subcellular distribution and activity of protein kinase C (PKC) is associated with transmembrane signalling in a variety of systems in which receptor occupancy leads to increased hydrolysis of polyphosphoinositides. Here we report evidence that in B lymphocytes, cyclic-cAMP-generating signal transduction pathways can activate translocation of PKC from the cytosol to the nucleus. Elevated cAMP levels and translocation of PKC to the nucleus are induced by antibodies against Ia antigens in normal B lymphocytes. Further, cAMP analogues mediate the translocation of PKC to the nucleus of these cells. These findings suggest that in physiological situations, ligation of B-lymphocyte Ia molecules by helper T cells leads to increased cAMP production which in turn causes PKC translocation to the nucleus. In view of recent observations that antibodies against Ia antigens induce differentiation of B cells, we conclude that nuclear PKC may function in the regulation of gene expression. PMID- 3037376 TI - Preferential relaxation of supercoiled DNA containing a hexadecameric recognition sequence for topoisomerase I. AB - In prokaryotes, the degree of supercoiling of DNA can profoundly influence the use of specific promoters. In eukaryotes, a variety of indirect observations suggest that DNA topology has a similar importance in proper gene expression. Much attention has therefore been focused on the cellular proteins that control DNA supercoiling, among which are the enzymes topoisomerase I and II. A hexadecameric sequence functions as a strong attraction site for topoisomerase I. Here we report that the interaction of topoisomerase I with this sequence motif is highly specific, because a single base-pair substitution prevents strand cleavage and thereby catalytic activity at the sequence. Thus, supercoiled DNA containing the recognition sequence is relaxed preferentially by topoisomerase I compared to a control, but no difference in the relaxation rate is observed for supercoiled DNA carrying the mutated sequence. The preference for the recognition sequence seems to be an intrinsic property of all eukaryotic type I topoisomerases, suggesting that the interaction might be important in a fundamental biological process. PMID- 3037377 TI - A new class of synthetic antibacterials acting on lipopolysaccharide biosynthesis. AB - Although there is a need for antibacterial agents that act only on Gram-negative bacteria, there are at present few such compounds. The 2-deoxy analogue of beta KDO (3-deoxy-beta-D-manno-2-octulopyranosonic acid) is a potent inhibitor of a key enzyme (CMP-KDO synthetase) in lipopolysaccharide biosynthesis of Gram negative bacteria, but it fails to penetrate intact bacteria. Coupling an L-L dipeptide to the 8-amino-2,8-dideoxy analogue of beta-KDO enabled it to be recognized and actively accumulated by certain peptide permeases of the cytoplasmic membrane. The dipeptide was hydrolysed in the cell and the inhibitor released. Subsequent inhibition of CMP-KDO synthetase led to the accumulation of large amounts of lipid A precursor and bacterial death. These compounds represent a new class of synthetic antimicrobials with a novel mechanism of action and considerable potential as chemotherapeutic agents. PMID- 3037378 TI - DNA chemistry. How the double helix breathes. PMID- 3037379 TI - Protein binding to DNA. PMID- 3037380 TI - A factor discriminating between the wild-type and a mutant polyomavirus enhancer. AB - Enhancers increase the frequency of transcription initiation from linked promoter elements, most probably as a result of the binding of specific proteins to the enhancer. The polyomavirus early region is expressed in differentiated mouse cells but not in undifferentiated embryonal carcinoma (EC) cells. This host range is a function of the enhancer because polyomavirus mutants selected for growth in EC cells have mutations in the enhancer and the host range is reproduced in transfection assays using the mutant enhancers. To understand the basis for this alteration in enhancer function, we have assayed extracts of EC cells for proteins that can interact with this sequence. We have detected a protein, present in a variety of cells, that can bind to the F441 mutant sequence, but binds only very poorly to the wild-type sequence. We conclude that this sequence alteration has probably generated a binding site for a positive-acting factor that allows the enhancer to function. PMID- 3037381 TI - A single mode of DNA base-pair opening drives imino proton exchange. AB - The opening of base pairs of double-stranded DNA is an important process, being a prerequisite for replication and transcription and possibly a factor in the recognition, flexibility and structure of DNA. The kinetics of base-pair opening have, however, been controversial. Base-pair opening can be studied by following the exchange of protons from imino groups with water, a process that seems only to occur from open base pairs. We have recently demonstrated catalysis by proton acceptors of imino proton exchange in nucleic acids. This has enabled us to determine the base-pair lifetimes, which are in the region of 10 ms at room temperature. In earlier reports it had been considered that proton exchange is limited by the rate of base-pair opening, which had led to estimates of base-pair lifetimes that were larger by one or two orders of magnitude. There are also important discrepancies between recent and early estimates of the base-pair dissociation constant. Earlier estimates of base-pair lifetimes correspond in fact to the time required for proton exchange in the absence of added catalyst (AAC exchange). This could be a distinct mode of base-pair opening with a very long open lifetime, different from the mode revealed by the effect of catalyst. The evidence reported here suggests on the contrary that there is only a single mode of here suggests on the contrary that there is only a single mode of base pair opening and that proton exchange in the absence of added catalyst is in fact catalysed by a proton acceptor intrinsic to the nucleic acid, most probably the other base of the open pair. PMID- 3037382 TI - Genetics of development elucidated by nematodes. PMID- 3037383 TI - The strychnine-binding subunit of the glycine receptor shows homology with nicotinic acetylcholine receptors. AB - We have cloned and sequenced cDNAs of the strychnine-binding subunit of the rat glycine receptor, a neurotransmitter-gated chloride channel protein of the CNS. The deduced polypeptide shows significant structural and amino-acid sequence homology with nicotinic acetylcholine receptor proteins, indicating that there is a family of genes encoding neurotransmitter-gated ion channels. PMID- 3037384 TI - Sequence and functional expression of the GABA A receptor shows a ligand-gated receptor super-family. AB - Amino-acid sequences derived from complementary DNAs encoding the alpha- and beta subunits of the GABA/benzodiazepine receptor from bovine brain show homology with other ligand-gated receptor subunits, suggesting that there is a super-family of ion-channel-containing receptors. Co-expression of the in vitro-generated alpha subunit and beta-subunit RNAs in Xenopus oocytes produces a functional receptor and ion channel with the pharmacological properties characteristic of the GABAA receptor. PMID- 3037385 TI - Induction of a factor that binds to the polyoma virus A enhancer on differentiation of embryonal carcinoma cells. AB - The cell type specificity of certain enhancers, competition experiments and the interactions that occur between proteins and enhancer sequences demonstrate that enhancers are the targets of specific factors involved in transcription control. The 246-base pair BclI-PvuII restriction enzyme fragment of polyoma virus has been shown to include two distinct enhancers, Py A and Py B, composed of several subdomains which interact with nuclear proteins from mouse fibroblasts. Embryonal carcinoma (EC) cells do not permit polyoma virus infection: both viral transcription and DNA replication are blocked. Host-range mutants of polyoma virus (EC mutants) capable of overcoming the expression block in EC cells have mutations or sequence rearrangements in their enhancer region. In an attempt to understand the molecular basis of this host restriction we compared the binding patterns displayed on the viral enhancer sequences by nuclear proteins prepared from EC cells or from fibroblasts. We show that one of the fibroblast factors required for Py A enhancer function, almost undetectable in EC cells, is induced after differentiation of these cells into parietal endoderm, suggesting that this protein is crucial in the regulation of viral gene expression during cellular differentiation, and perhaps more generally in the control of gene expression during early embryonic development. PMID- 3037386 TI - Dual roles for DHP receptors in excitation--contraction coupling? PMID- 3037387 TI - Primary structure of the receptor for calcium channel blockers from skeletal muscle. AB - The complete amino-acid sequence of the receptor for dihydropyridine calcium channel blockers from rabbit skeletal muscle is predicted by cloning and sequence analysis of DNA complementary to its messenger RNA. Structural and sequence similarities to the voltage-dependent sodium channel suggest that in the transverse tubule membrane of skeletal muscle the dihydropyridine receptor may act both as voltage sensor in excitation-contraction coupling and as a calcium channel. PMID- 3037388 TI - Expression and function of CD4 in a murine T-cell hybridoma. AB - The CD4 (T4) antigen was originally described as a phenotypic marker specific for helper T cells, and has recently been shown to be the receptor for the human immunodeficiency virus (HIV). Functional studies using monoclonal antibodies directed at CD4 and major histocompatibility complex (MHC) class II molecules led to the suggestion that CD4 binds to the MHC class II molecules expressed on stimulator cells, enhancing T-cell responsiveness by increasing the avidity of T cell-stimulator cell interaction and/or by transmitting a positive intracellular signal. But recent evidence that antibodies to CD4 inhibit T-cell responsiveness in the absence of any putative ligand for CD4 has been interpreted as suggesting that antibody-mediated inhibition may involve the transmission of a negative signal via the CD4 molecule instead. We have infected a murine T-cell hybridoma that produces interleukin 2 (IL-2) in response to human class II HLA-DR antigens with a retroviral vector containing CD4 cDNA. The resulting CD4-expressing hybridoma cell lines produce 6- to 20-fold more IL-2 in response to HLA-DR antigens than control cell lines. Furthermore, when antigen levels are suboptimal, the response of the cell lines is entirely CD4-dependent. The data presented here clearly demonstrate that CD4 can enhance T-cell responsiveness and may be crucial in the response to suboptimal levels of antigen. PMID- 3037389 TI - Beta-endorphin-sensitive opioid receptors in the rat tail artery. AB - Isolated tail arteries of rats were perfused and field-stimulated every 2 min with 2 pulses at 1 Hz. Different opioid peptides depressed the contractile responses to stimulation; their concentration-response curves showed a maximum at about 40% inhibition. The rank order of potency of the peptides was beta endorphin (IC50 = 97 nmol/l) approximately equal to BAM-22P greater than FK-33824 greater than DAGO greater than [D-Ala2,D-Leu5]-enkephalin greater than or equal to metorphamide greater than dynorphin A-(1-13) approximately equal to [Met5]enkephalin. All these substances have in common a certain activity at opioid mu-receptors, although the enkephalins are preferential delta-, and the dynorphins preferential kappa-agonists. However, the selective delta-agonist [D Pen2,L-Pen5]enkephalin was ineffective at up to 10 mumol/l, and the kappa agonists ethylketocyclazocine and U-50488 acted only at concentrations higher than 3 mumol/l. Whereas the effects of beta-endorphin, DAGO and [D-Ala2,D Leu5]enkephalin could be reduced by the mu-preferential antagonist naloxone, the effects of ethylketocyclazocine and U-50488 were not changed. The delta-selective antagonist ICI 174864 did not influence the action of [D-Ala2,D-Leu5]enkephalin. Naloxone in a concentration (1 mumol/l) which nearly abolished the effect of DAGO 3 mumol/l, slightly enhanced responses to stimulation. Neither beta-endorphin nor DAGO influenced vasoconstriction evoked by the application of noradrenaline or adenosine triphosphate; U-50488 reduced it. In arteries preincubated with [3H]noradrenaline DAGO depressed, whereas naloxone enhanced the tritium overflow and vasoconstriction evoked by field stimulation (0.4 Hz, 24 pulses every 14 min). In addition, naloxone antagonized the effect of DAGO.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3037390 TI - Plasma concentration and vascular effect of beta-endorphin in spontaneously hypertensive and Wistar Kyoto rats. AB - In order to find out whether beta-endorphin (beta-E) is involved in the development of hypertension, we performed two series of experiments. Firstly, spontaneously hypertensive rats (SHR) and their normotensive Wistar Kyoto controls (WKY) were submitted to ether stress. Plasma concentrations of beta endorphin-like immunoreactivity (beta-EI), adrenocorticotropin (ACTH) and alpha melanotropin (alpha-MSH) were measured by radioimmunoassay. The basal concentration of beta-EI was similar in WKY and SHR, whereas WKY had higher levels of ACTH and lower levels of alpha-MSH than SHR. In both strains acute stress enhanced the plasma concentration of beta-EI to the same extent and with a similar time-course. The increase of plasma beta-EI coincided with a rise in ACTH but not alpha-MSH. Gel chromatography of beta-EI revealed that plasma extracts contain similar amounts of beta-lipotropin- (beta-LPH) and beta-E-sized immunoreactive components, and that acute stress elevated both forms of beta-EI. Secondly, isolated tail arteries of SHR and WKY were perfused and field stimulated with two pulses at 1 Hz. beta-E depressed stimulation-evoked vasoconstriction with the same potency in both strains. Thus, basal and stress induced levels of beta-EI did not differ in SHR and WKY. Moreover, in the tail artery of both strains the sensitivity of presynaptic opioid receptors towards beta-E was almost identical. If the beta-E sensitivity of these receptors in other arteries of WKY and SHR is also similar a major role of the circulating peptide in the development of hypertension is rather unlikely. PMID- 3037391 TI - Analysis of the mechanism of action of bradykinin on human basilar artery in vitro. AB - Bradykinin (BK) initially produced concentration-related relaxations of human basilar artery in vitro. Concentration-effect curves constructed at 2 h intervals to BK over an 8 h period were reproducible. The rank order of potency of three kinins on the human basilar artery was found to be BK greater than methionyl lysyl-BK greater than des-Arg9-BK. The B2-receptor antagonist Thi5,8, D-Phe7-BK but not the B1-receptor antagonist des-Arg9-Leu8-BK selectively blocked BK induced relaxations of the human basilar artery. The relaxant effects of bradykinin and acetylcholine but not papaverine were attenuated after removal of the endothelium or treating the tissues with BW755C. Indomethacin was without effect. Concentration-effect curves to angiotensin I were markedly attenuated by captopril at a concentration which had no effect on BK, angiotensin II or 5 hydroxytryptamine responses. It is concluded that BK induced relaxations of the human basilar artery are mediated via activation of a B2 receptor and the response is dependent upon the release of a factor present in the endothelium. Angiotensin converting enzyme is present in the human basilar artery and is important for the conversion of angiotensin I to angiotensin II but apparently not for the degradation of BK. It is likely that other kininases are present and active in the tissue. PMID- 3037392 TI - Evidence against a role of a pertussis toxin-sensitive guanine nucleotide-binding protein in the alpha 1-adrenoceptor-mediated positive inotropic effect in the heart. AB - Pertussis toxin, which specifically inactivates guanine nucleotide-binding proteins (N-proteins) involved in the signal transduction in various receptor systems, did not influence the positive inotropic effect of the alpha 1 adrenoceptor agonist phenylephrine in rat isolated left auricles. This indicates that the alpha 1-adrenoceptor-mediated positive inotropic effect does not involve a pertussis toxin-sensitive N-protein. PMID- 3037393 TI - [Azidothymidine and HIV infection]. PMID- 3037394 TI - [The treatment of chronic cardiac decompensation]. PMID- 3037395 TI - [Changes in presynaptic processes during tonic subthreshold activation of the spinal center of scratching movements in the cat]. AB - In immobilized intercollicularly decerebrated cats tonic underthreshold activation of the spinal scratching generator (after application of tubocurarine or bicuculline on C1-C2 segments) is accompanied by an increase in primary different terminal depolarization, decrease in N1-component of cord dorsum potential evoked by stimulation of cutaneous afferents, decrease in the amplitude of DRP and early polysynaptic responses of motoneurons evoked by stimulation of cutaneous and muscle afferents; a respective rise and reduction in activity of intermediate nucleus interneurons which are mono- and di (oligo)-synaptically connected with afferent terminals. Spinalization of animal led to reverse changes. Injection of DOPA into spinal animals allowed comparing changes in the state of lumbar segmental apparatus during tonic underthreshold activation of spinal scratching and locomotor generators. PMID- 3037396 TI - Lung carcinoma in uranium miners, Czechoslovakia, 1976-1980. AB - The analysis of the clinical data of uranium miners suffering from lung cancer in the years 1976-1980 was made. In 299 diseased men with lung cancer verified by histology and/or cytology the average age was 56.2 years. There were 52.8% of epidermoid carcinomas, 32.8% of small cell carcinomas, 5% of adenocarcinomas, and 9.4% of other, mixed, undifferentiated carcinomas. This distribution differed from those reported previously. In 25 survivors of 5 years (8.4%), there were 21 patients radically operated in the Stage I or II of the disease. In 84% of survivors the cancer was epidermoid. The lung cancer in uranium miners has not any proper characteristics excluding the age of diseased persons which is about 10-15 years lower than in current male population suffering from lung cancer. PMID- 3037397 TI - [Simultaneous occurrence of brain tumors and aneurysms]. AB - Three patients with different types of intracranial tumour--glioma, meningioma and pituitary adenoma--and associated aneurysms are described. The aetiology of aneurysm formation in the presence of tumours is discussed and a review of the literature given. PMID- 3037398 TI - [Bilateral symmetrical reversible thalamic lesion within the scope of encephalitis]. PMID- 3037399 TI - [Unilateral sensory neuropathy of the trigeminal nerve as the leading symptom of primary Sjogren syndrome ("sicca syndrome")]. AB - We report a case who presenting with a progressive numbness and mild hyperpathia in the second cutaneous division of the left trigeminal nerve as the leading symptom of Primary Sjogren's Syndrome. Further typical features of this autoimmune disorder are keratoconjunctivitis sicca and xerostomia. A symmetrical, predominantly sensory polyneuropathy can be revealed by sensory nerve conduction studies of the median and sural nerves. Laboratory findings mostly include elevated erythrocyte sedimentation rate, hypergammaglobulinemia and hypercomplementemia. The presence of the precipitating antinuclear antibodies SS A and/or SS-B is pathognomonic for Sjogren's Syndrome. As long as the disease remains benign, treatment should be symptomatic. Malignant exacerbations require immunosuppressive treatment. PMID- 3037400 TI - Plasma levels of main granulocyte components during hemodialysis: effects of immunosuppression. AB - Plasma levels of granulocyte lactoferrin, granulocyte myeloperoxidase and granulocyte elastase in complex with alpha 1-proteinase inhibitor were investigated during hemodialysis in oliguric patients following cadaveric renal transplantation (CRT). The results were compared with those of patients undergoing regular hemodialysis treatment (RDT) using different membrane materials. In RDT patients, plasma lactoferrin levels increased from 61.9 +/- 10.2 to 417.9 +/- 96.7 using dialyzers made of cuprophane. Dialyzers made of polymethyl methacrylate induced an increase of lactoferrin from 166.6 +/- 28.5 to 712.5 +/- 165.9 and dialyzers made of polyacrylonitrile an increase from 122.6 +/ 23.5 to 647.7 +/- 203.6 ng/ml. In CRT patients, in contrast, plasma lactoferrin levels increased from 49.1 +/- 10.9 to 199.6 +/- 45.9 (cuprophane), from 30.1 +/- 6.9 to 252.5 +/- 46.9 (polymethyl methacrylate), and from 43.3 +/- 9.1 to 174.2 +/- 39.3 ng/ml (polyacrylonitrile). On the other hand, the plasma levels of myeloperoxidase and elastase in complex with alpha 1-proteinase inhibitor increased comparably in both groups of patients. Our data suggest that immunosuppression might prevent degranulation of specific granules independently of the used dialyzer membrane material, whereas no effect was observed on two main components of azurophilic granules. PMID- 3037401 TI - Behavioral changes in ACTH-(1-24)-induced excessive grooming in aging rats. AB - Age-related behavioral changes are commonly associated with impaired cognitive functioning. As a result an abundance of neuropsychological tools have been developed to study learning and memory in man and animal. The combined use of ethological methods and the extensively studied measures of excessive grooming behavior in rats provides an excellent tool to study non-cognitive behavioral changes. In the present study, a sequential analysis was applied to the different grooming elements. Aged rats showed a loss in the sequential organization of their grooming behavior, which led to a decrease in the display of tail sniffing. Interestingly, the reduced display of an element which appears latest in the ontogeny of grooming is reminiscent of the Jacksonian principle of behavioral degeneration. Seemingly, this principle is not necessarily confined to learning paradigms, but may be extended to other behavioral systems as well. PMID- 3037402 TI - Properties of the 3'-phosphoadenosine-5'-phosphosulfate (PAPS) synthesizing systems of brain and liver. AB - Chromatography of brain and liver 100,000 g supernatants over HPLC molecular sieve columns revealed striking differences in the molecular weight distribution of ATP-sulfurylase and APS-kinase of the two tissues, pointing to different enzymic species for both enzymes in brain and liver. This was further substantiated by kinetic characterization of the two enzymes of both tissues. APS kinase of liver is allosterically activated by ATP, while the brain enzyme is not. ATP-sulfurylase of brain is activated at high, but still physiological concentrations of ATP. Brain ATP-sulfurylase is inhibited by phenylalanine. PMID- 3037403 TI - Detection of choline kinase in purified rat brain myelin. AB - Choline kinase, an enzyme involved in the Kennedy pathway conversion of diacylglycerol to phosphatidylcholine, was detected in highly purified rat brain myelin at a level equal to 20% that of whole brain homogenate. This was an order of magnitude higher than the specific activity of lactate dehydrogenase, marker for cytosol. Choline kinase was also detected in the P1, P2, P3, and cytosolic fractions with highest relative specific activity in the latter. Myelin washed with buffered sodium chloride or taurocholate retained most of its kinase, indicating that adsorption of the soluble enzyme was unlikely. The results of mixing experiments and repeated purification further indicated that the enzyme is intrinsic to myelin. This finding in concert with previous studies supports the concept that myelin has all the enzymes needed to convert diacylglycerol to phosphatidylcholine. PMID- 3037404 TI - Ethanolamine base exchange in astrocyte primary cultures: localization and developmental studies. AB - The enzymatic activities of ethanolamine base exchange (EBEE) and CDP ethanolamine: 1,2-diacylglycerol ethanolamine phosphotransferase (EPT) were investigated during the growth of rat astrocyte primary cultures. From the 16th day, cells ceased to divide (2.0 X 10(6) cells per culture dish); the total phospholipid (PL) content increased 1.5 fold between the 16th and 24th day (0.20 to 0.30 mumol per mg protein) but the amount of ethanolamine phospholipid (28% of PL content) remained constant. Whereas the specific activity (pmol/min X mg protein) of EPT reached a plateau at 16 days in culture and remained constant (400) thereafter, that of EBEE increased up to the 19th day (190) and decreased gradually to a basal level (75) at the 24th day. EBEE activity was not detected in plasma membranes isolated from 16, 19 and 24 days astrocyte cultures. Sub cellular fractionation and determination of EBEE specific activities showed that the 104 X 10(3) g fraction (P4) was 4.8 and 8.8 fold enriched at the 16th day and 24th day respectively as compared to the whole cell homogenate (50 and 75). The 7 X 10(3) g (P2) and 17 X 10(3) g (P3) fractions were 8.4 and 7.0 fold enriched respectively at the 19 day in culture. The percentages of the enzymatic activity in the different subcellular fractions were 30, 57.2 and 25.7 for P2 and 39.2, 2.6 and 39.8 for P4 at 16, 19 and 24 days in culture respectively. The activity remained constant in P3 (23%) and was negligible in P1 (6%). Ultrastructural studies revealed that P2 and P3 were enriched in mitochondria while P4 contained essentially microsomes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3037405 TI - Cyclic nucleotides and retinal cones. PMID- 3037406 TI - Fluoroenzymatic cycling assay (FECA) for the determination of catechol estrogen monomethyl ethers in human urine. AB - In the present study a method is described for the quantitative determination of the methylated metabolites of catechol estrogens in human urine. Following initial enzymatic hydrolysis the urine samples are extracted with ethyl acetate. The monomethyl ethers of catechol estrogens are then selectively fractionated with straight phase chromatography on Lipidex-5000 gel. Finally, samples are quantitated using enzymatic cycling with 17 beta-estradiol dehydrogenase combined with fluorometry. The method is sensitive, reproducible and reasonably rapid for routine analysis and avoids the hazards of radioisotopes. Preliminary values of normal males and non-pregnant females are presented. PMID- 3037407 TI - Vanadate inhibition of rat cerebral cortex Na+,K+-ATPase during postnatal development. AB - The Na+,K+-ATPase activity and its response to vanadate inhibition was investigated in cerebral cortex homogenates of 7-, 12- and 18-day-old rats. The enzyme was inhibited by vanadate in a dose-dependent manner in all these age groups. Furthermore, there was a different sensitivity towards vanadate during postnatal development; the concentration of V+5 needed for 50% inhibition of Na+, K+-ATPase was 1.1 X 10(-6)M, 2 X 10(-7)M and 4.4 X 10(-7)M for 7-, 12- and 18-day old rats, respectively. It is suggested that the different sensitivity of Na+, K+ ATPase towards vanadate inhibition during postnatal development might be due to age-dependent changes in the ratio of various cell types. PMID- 3037408 TI - [IgG synthesis in the cerebrospinal fluid in patients with the chronic progressive phase of multiple sclerosis treated with cyclophosphamide and ACTH]. AB - In 30 cases of multiple sclerosis in the chronic-progressive phase treated with cyclophosphamide (total dose 4000 mg) and ACTH (total dose 1050 units) IgG and albumin were determined in the serum and cerebrospinal fluid by Mancini's method. From these determination the IgG, albumin ratio and the IgG index were calculated as markers of IgG synthesis in CSF spaces. As a result of these investigations it is suggested that the treatment of patients with multiple sclerosis in chronic progressive phase with short-lasting intensive immunosuppression using cyclophosphamide and ACTH causes no significant changes in IgG synthesis in CSF spaces. PMID- 3037409 TI - [ACTH in the plasma and cerebrospinal fluid in patients with multiple sclerosis]. AB - In 1984 and at the beginning of 1985 the authors carried out radioimmunoassays (SORIN-CIS kit) the plasma levels of ACTH in 116 multiple sclerosis patients (m 52, f-64) and in 10 cases this radioimmunoassay was done in the cerebrospinal fluid (m-5, f-5). The control group comprised 90 patients with ischialgia and neuroses. The normal value in the plasma was from 0 to 80.86 pg/ml, and in the fluid it was from 0 to 77.08 pg/ml. In multiple sclerosis patients the plasma ACTH level was from 0 to 286.9 pg/ml, in the cerebrospinal fluid from 0 to 89 pg/ml. The values of ACTH were significantly higher in multiple sclerosis patients, mainly in males. In the fluid the level of ACTH was significantly higher in the studied patients. No significant differences in ACTH levels were found between males and females with multiple sclerosis, and in the control group this level was higher in females. Raised ACTH level was found mainly in multiple sclerosis with lung duration of the disease (10 years) at the time of exacerbations. The authors continue studies on the axis hypothalamus-hypophysis adrenals, on various hormones, prostaglandins, beta-endorphin, biochemical markers, cAMP, cCMP, arylosulphatase A and B MBC etc. PMID- 3037410 TI - Mental retardation associated with elevated cerebrospinal fluid antibody levels to herpes simplex, adeno and mumps viruses. PMID- 3037411 TI - Effects of the active constituents of Catha edulis on the neuromuscular junction. AB - (-)-Cathinone and d-norpseudoephedrine (DNE) in the dose range 0.2-1.2 mg/ml produced a reduction in contractions of skeletal muscle, evoked by direct and indirect electrical stimulation and antagonised the facilitatory action of physostigmine on the neuromuscular junction; but failed to antagonise a partial blockade induced by d-tubocurarine (dTb) as occurs with norepinephrine or epinephrine. The local anaesthetic actions of (-)-cathinone and DNE were found to be almost equivalent to that of lignocaine. These results indicate that (-) cathinone and DNE may have a direct blocking action on the neuromuscular junction, which is independent of cholinergic and adrenergic transmission. PMID- 3037412 TI - Defense reaction elicited by injection of GABA antagonists and synthesis inhibitors into the posterior hypothalamus in rats. AB - Blockade of gamma-aminobutyric acid (GABA) in the posterior hypothalamic nucleus elicits cardiorespiratory stimulation in anesthetized rats. The present study was conducted to test the hypothesis that blockade of GABA in this cardiostimulatory area of the posterior hypothalamus in conscious animals would elicit a defense reaction characterised by a "fight or flight" response. Blockade of GABA was achieved by injecting bicuculline methiodide (BMI 1-25 ng) and picrotoxin (4-100 ng), two post-synaptic GABA antagonists and isoniazid (INH 35 and 70 micrograms), an inhibitor of the synthesis of GABA, bilaterally into the posterior hypothalamus through chronically implanted microinjection cannulae. All three drugs produced dose-dependent increases in locomotor activity, suggesting an "escape" reaction which was quantified as number of crossings and rearings. The effects of bicuculline and picrotoxin appeared immediately after the injection while those of isoniazid appeared much more slowly, attaining peak effects 24 +/- 1 min after injection. Injection of either strychnine (38 ng) into the posterior hypothalamus or bicuculline into the lateral hypothalamic area (LHA) or the dorso medial/ventro-medial hypothalamus (DMH/VMH) did not elicit a significant increase in locomotor behavior. These results suggest that both the physiological and locomotor components of the hypothalamic defense reaction may be under tonic GABAergic inhibition in the region of the posterior hypothalamus. PMID- 3037413 TI - Cardiorespiratory effects produced by injecting drugs that affect GABA receptors into nuclei associated with the ventral surface of the medulla. AB - It has recently been shown that L-glutamic acid induced stimulation of cell bodies in a circumscribed area of the rostral ventrolateral medulla (RVLM) in the cat, produced increases in arterial pressure (AP), decreases in heart rate (HR) and transient apnea (Gatti, Norman, DaSilva and Gillis, 1986). The purpose of the present study was to determine if this same area was sensitive to GABA receptor agonists and antagonists. Injection of the GABA agonist muscimol (200 ng), into the rostral ventrolateral medulla of cats anesthetized with chloralose produced a precipitous and immediate fall in arterial pressure (-95 +/- 4.6) and heart rate (-31 +/- 5.9; n = 4, P less than 0.05). Maximal cardiovascular effects could only be achieved if muscimol was injected bilaterally. These effects of muscimol on arterial pressure were dose-dependent. Time-action curves for the effects of muscimol on arterial pressure and respiration were different. Hypotension occurred first and was followed later in time by a decrease in minute ventilation. Within 30 min all animals were apneic after the 200 ng dose. The cardiovascular effects of muscimol were reversed by the injection of the GABA receptor antagonist bicuculline. These data indicate that stimulation of GABA receptors in the rostral ventrolateral medulla produced selective cardiovascular effects and that respiratory neurons sensitive to GABA are apparently not localized with these cardiovascular neurons. PMID- 3037414 TI - The GABA agonist THIP, attenuates antinociception in the mouse by modifying central cholinergic transmission. AB - The effect of THIP, a direct-acting gamma-aminobutyric acid (GABA) receptor agonist, on the antinociceptive response to a variety of agents was examined using the mouse tail-immersion assay. Alone, THIP produced an antinociceptive response in smaller doses (5 mg/kg) but was ineffective at doses exceeding 10 mg/kg. Treatment with THIP (15 mg/kg) was found to block the antinociceptive response to an inhibitor of the uptake of GABA, an inhibitor of GABA transaminase, a direct-acting GABA receptor agonist and to a cholinesterase inhibitor. In contrast, THIP had no effect on the antinociceptive responses to morphine, clonidine or oxotremorine. The results indicate that large doses of THIP reduce cholinergic activity in a pathway important for mediating the antinociceptive action of GABAergic drugs and physostigmine. PMID- 3037415 TI - Effects of trifluoperazine on the cholinergic function of the hippocampus of the rat. AB - The changes induced in the level and turnover rate (TRACh) of acetylcholine (ACh) in the hippocampus and in the in vitro release of [3H]ACh by intracerebroventricular injection of trifluoperazine (TFP) were studied. Rats were killed by microwave irradiation at various times after treatment with drug or vehicle and the levels of ACh were measured by gas chromatography. After the administration of 100 micrograms trifluoperazine, the content of ACh of the whole hippocampus increased from 24.0 +/- 2.9 to 35.2 +/- 4.2 nmols/g wet weight over a 60 min period, while the turnover rate was markedly decreased (from 0.86 to 0.53 nmols/min/g wet tissue weight). The potassium-evoked release of [3H]ACh from slices of hippocampus under in vitro conditions was decreased in a concentration dependent manner by preincubation with 50 and 100 microM of trifluoperazine for 60 min. The results support the assumption that trifluoperazine can alter neurotransmission, possibly by inhibiting the calcium-binding protein calmodulin, and/or by interfering with cell metabolism. PMID- 3037416 TI - Differential tolerance to repeated daily injections of N-allylnormetazocine and its enantiomers in the rat. AB - This study was designed to compare the development of tolerance to the effects of N-allylnormetazocine (SKF-10,047) and its enantiomers on the EEG and on behavior. Adult female Sprague-Dawley rats were implanted with chronic cortical electroencephalogram (EEG) and electromyogram (EMG) recording electrodes in the temporalis muscle and with permanent cannulae in the external jugular vein. In non-tolerant rats, 10 mg/kg (i.v.) injections of SKF-10,047 racemate produced primarily aroused wakefulness for about 120 min, that was associated with alternation between desynchronized EEG and theta waves in the EEG. After these rats received a series of automatic, intravenous injections of SKF-10,047 racemate, the aroused wakefulness state induced by SKF-10,047 racemate lasted for about 40 min. In non-tolerant rats, 2.5 mg/kg (i.v.) injections of (+)-SKF-10,047 induced a psychotomimetric EEG and behavioral state for about 30 min, which included continuous theta wave activity in the EEG. After chronic treatment with (+)-SKF-10,047, the psychotomimetic state induced by (+)-SKF-10,047 persisted for about 20 min. In non-tolerant rats, 2.5 mg/kg (i.v.) injections of (-)-SKF-10,047 produced an aroused EEG and behavioral wakefulness for about 30 min, which was then followed by slow-wave bursts in the EEG and associated behavioral stupor for about 90 min. After chronic treatment with (-)-SKF-10,047, injection of (-)-SKF 10,047 produced predominantly aroused wakefulness for about 45 min. The data suggest that (+)-SKF-10,047 exerts psychotogenic properties, but not opioid properties. On the other hand, the data suggest that (-)-SKF-10,047 possesses opioid properties. PMID- 3037417 TI - Actions of opioids on the dorsal root potential of the isolated spinal cord preparation of the neonate rat. AB - The effects of opioids were studied on the dorsal root-dorsal root evoked potential (DRP) of the neonatal hemisected spinal cord of the rat in vitro. Applications of [D-Ala2,Met5]enkephalinamide (DAME), [D-Ala2,D-Leu5]enkephalin (DADL), Leu5 enkephalin, Met5 enkephalin, Dynorphin 1-9 and normorphine produced dose-dependent depressions of the dorsal root potential. The depressant effects of these agents were antagonised by naloxone but not by the highly selective delta-opioid receptor antagonist ICI 174864. Compounds acting on the kappa opioid receptor had only weak and inconsistent inhibitory effects on the dorsal root potential. Morphine had antagonist properties on the dorsal root potential, as did the kappa agonists ethylketocyclazocine and bremazocine, but not U50488 and tifluadom. The depressant actions of opioids on the dorsal root potential thus appeared to be mediated by actions on the mu receptor type. However, only mu agonists with relatively high intrinsic activity were able to reduce the dorsal root potential. Other mu agonists, including morphine, acted as antagonists. Actions on receptors for the delta and kappa agonists, in contrast, appeared to have little influence on this response. The antagonist actions of certain kappa agonists were probably due to their high affinity for, but low efficacy at mu receptors. PMID- 3037418 TI - 1-Methylisoguanosine: interaction with central adenosine receptors and lack of antagonism of its in vivo effects by a benzodiazepine antagonist. AB - 1-Methylisoguanosine, a marine natural product analogue of adenosine, with moderate activity as a benzodiazepine receptor ligand, has previously been shown to have muscle-relaxant and hypothermic activity in mice in vivo. The present experiments showed that the benzodiazepine antagonist Ro15-1788 did not block the in vivo muscle-relaxant and hypothermic effects of 1-methylisoguanosine, suggesting that these particular actions are not due to interactions with benzodiazepine receptors. When applied by microiontophoresis near spontaneously active neurones or neurones activated by ACh, DL-homocysteate or glutamate in the ventrobasal thalamus of anaesthetized rats, 1-methylisoguanosine had a depressant action; it was similar to adenosine in potency and in its ability to be antagonized by 8-(parasulphophenyl)theophylline. The depression was usually longer lasting than that caused by adenosine, consistent with previous neurochemical data showing it to be resistant to adenosine deaminase and a poor substrate for the uptake system for adenosine in the CNS. These results suggest that the major pharmacological/behavioural actions of 1-methylisoguanosine in vivo are more likely to be caused by an interaction with adenosine receptors, rather than with benzodiazepine sites. PMID- 3037419 TI - Discriminative stimulus properties of chlordiazepoxide and zolpidem. Agonist and antagonist effects of CGS 9896 and ZK 91296. AB - In previous studies the effects of CGS 9896, a pyrazoloquinoline ligand at benzodiazepine receptors, in rats trained to discriminate benzodiazepines from vehicle, have been variable. The present experiment confirmed that this compound produced responding on the drug-lever in rats trained to discriminate 5 mg/kg of chlordiazepoxide from saline, and showed that CGS 9896 did not antagonise the effect of chlordiazepoxide in this test. In contrast, CGS 9896 antagonised the stimulus properties of zolpidem (2 mg/kg), a non-benzodiazepine hypnotic, which displaces benzodiazepines from their binding sites. The drug CGS 9896 also antagonised responding on the drug-lever produced by chlordiazepoxide in rats trained with zolpidem. The beta-carboline, ZK 91296, produced effects similar to those of CGS 9896, giving rise to responding on the drug-lever in rats trained with chlordiazepoxide and antagonising the zolpidem cue. These results demonstrate the mixed agonist-antagonist effects of CGS 9896 and ZK 91296 and suggest that the stimulus properties of chlordiazepoxide and zolpidem may be mediated by different sub-types of benzodiazepine receptors. PMID- 3037420 TI - Depolarisation-evoked release of acetylcholine can mediate phosphoinositide hydrolysis in slices of rat cerebral cortex. AB - Depolarisation of [3H]inositol prelabelled slices of cerebral cortex of the rat, with elevated extracellular K+ or the alkaloid veratrine, induced a marked accumulation of [3H]inositol monophosphate in the presence of 5 mM Li+. The effects of these stimuli were concentration-related with maximal responses obtained at 30 mM K+ and 30 microM veratrine. Larger concentrations produced submaximal responses but also markedly suppressed the incorporation of [3H]inositol into phospholipid. The responses to K+ or veratrine were not sensitive to atropine, prazosin, mepyramine, ketanserin or the peptidase inhibitor bacitracin. However, in the presence of the cholinesterase inhibitor physostigmine, the responses to these stimuli were greatly enhanced and this could be blocked by atropine. Both veratrine and K+ markedly stimulate release of endogenous acetylcholine from the slices. Release appears to be linear with time over the 45 min period of continuous stimulation. Reduction of extracellular calcium severely suppressed both the release of acetylcholine and the atropine sensitive component of the phosphoinositide response to K+. The results suggest that endogenous acetylcholine can stimulate phosphoinositide metabolism by interacting with muscarinic receptors. The atropine-insensitive component, at least in part, represents entry of Ca2+ through voltage-sensitive channels and perhaps a direct effect on phosphoinositide metabolism. PMID- 3037421 TI - Opioid receptors and neuropeptides in the CNS in rats treated chronically with amoxapine or amitriptyline. AB - The central mechanism responsible for the potentiation by antidepressant drugs of analgesia induced by morphine, was explored by measuring the levels of various neuropeptides (met-enkephalin, leu-enkephalin, dynorphin, substance P and cholecystokinin-like materials) and the density of delta and mu opioid binding sites in the spinal cord of rats treated for 14 days with amoxapine (10 mg/kg i.p., daily) or amitriptyline (10 mg/kg i.p., daily). Similar measurements were made in the hypothalamus and cerebral cortex for comparison. Chronic treatment with amoxapine or amitriptyline did not affect the levels of dynorphin, substance P and cholecystokinin, but markedly enhanced the levels of leu-enkephalin in the three structures examined. The levels of met-enkephalin were also increased after treatment with amitriptyline but only in the spinal cord and hypothalamus. No changes in opioid receptors were found in the cerebral cortex, but the densities of delta and mu opioid binding sites were increased in the spinal cord, and decreased in the hypothalamus of rats treated with amoxapine or amitriptyline. These changes induced by antidepressants in opioidergic markers at the spinal level might account for the potentiation of the action of morphine in amoxapine- or amitriptyline-treated rats. In addition, the observed alterations in the same markers in the hypothalamus could be associated with changes induced by antidepressants in neuroendocrine regulation. PMID- 3037422 TI - Binding sites for a peripheral type benzodiazepine antagonist ([3H]PK 11195) in human iris. AB - Peripheral-type benzodiazepine binding sites have been characterized on sections of 8 normal human iris/ciliary-body preparations. Saturability was determined at 25 degrees C with [3H] PK 11195 (1 nM) a specific ligand of peripheral type sites. The studies revealed a single class of binding sites for PK 11195 with a nanomolar range affinity (KD = 1.45 nM) and a maximal capacity (Bmax) of 35.5 fmol/mg protein. The displacement potency order of the benzodiazepines tested suggest that these sites belong to the peripheral type: PK 11211 (IC50 = 12 nM) greater than Ro 5-4864 (IC50 = 770 nM) greater than clonazepam (IC50 = 20,000 nM). The present data demonstrate that high affinity binding sites for peripheral type benzodiazepines are present in human iris/ciliary-body. This tissue is therefore a suitable tool for evaluation of the putative functional role of these binding sites. PMID- 3037423 TI - Desipramine and noradrenergic neurotransmission in aging: failure to respond in aged laboratory animals. AB - Deficiencies in noradrenergic neurotransmission have been found in the central nervous system of aged laboratory animals. The purpose of the present study was to determine if tricyclic antidepressants, such as desipramine, can overcome the diminished noradrenergic neurotransmission found in these animals. Using electrophysiological techniques, noradrenergic neurotransmission was examined in the cerebellar cortex of rats, a model system which has been used extensively to characterize the effects of norepinephrine in the central nervous system. The discharge rate of cerebellar Purkinje neurons is very sensitive to changes in the noradrenergic input from the nucleus locus coeruleus. In this model system in young rats, treatment with desipramine slowly augments noradrenergic neurotransmission over several weeks. Similar treatment in aged animals caused no increase in the age-related deficient noradrenergic neurotransmission. The decline in efficacy of desipramine with age could not be accounted for by differences between young and old rats in the distribution of the drug. Failure of desipramine to be effective in older rats may reflect the insensitivity of aged neurons to norepinephrine itself, so that treatment strategies which increase the amount of nerepinephrine released onto these neurons may be ineffective. The findings may have implications for the use of tricyclic antidepressants in aged depressed patients. PMID- 3037424 TI - Effect of mono- and diaminopyridines on release of [3H]norepinephrine from isolated guinea-pig atrium. AB - Neurochemical evidence has been obtained that 4-aminopyridine, 3,4 diaminopyridine and 3,3-dimethyl-1-(4-amino-3-pyridyl)urea HBr (LF-14), concentration-dependently enhanced the stimulation-evoked release of [3H]norepinephrine ([3H]NE) from isolated guinea-pig atrium. The effects of aminopyridines, compounds known to inhibit potassium channels, were Ca0 dependent. High pressure liquid chromatography, combined with radiochemical detection, indicated that the increased stimulated release of radioactivity was due to [3H]NE. Since the aminopyridines studied also enhanced the release of [3H]NE from atrium treated with cocaine, a blocker of uptake1, it seems likely that the increased release of NE caused by the aminopyridines is due to the enhanced release of NE from sympathetic axon terminals and not to the inhibition of reuptake. It is probable that the sympathomimetic cardiac effects (positive inotropic and chronotropic effect) of aminopyridines observed in animal experiments is due to the increased release of NE, caused by these compounds. PMID- 3037425 TI - Effects of chronic administration of dextroamphetamine on enzymes of energy metabolism in regions of the rat brain. AB - In the present study the effects of chronic administration of dextroamphetamine on energy metabolism in the brain of the rat were examined. The enzymes studied were: hexokinase (soluble and particulate forms), phosphofructokinase, pyruvate kinase, lactate dehydrogenase, citrate synthase, NAD+ and NADP+-dependent isocitrate dehydrogenases, succinate dehydrogenase and malate dehydrogenase. All the activities of the enzymes were assayed in four regions of the brain of the rat (cerebellum, medulla oblongata and pons, cererbral cortex and diencephalon). Rats were injected intaperitoneally once daily with dextroamphetamine for 20 consecutive days. The initial dose was 5 mg/kg/day and the dose was then increased by 1 mg/kg/every 5 days up to a total of 8 mg/kg/day on days 16-20. In the glycolytic enzymes a reduction of the activity of phosphofructokinase was found in the diencephalon and an increase of the activity of pyruvate kinase and lactate dehydrogenase in the diencephalon and medulla oblongata and pons, respectively. Citrate synthase was the only enzyme in the Krebs' cycle affected by chronic administration of dextroamphetamine. The results presented here show that chronic administration of dextroamphetamine produced important changes in some enzymes of glycolysis and the Krebs' cycle in the brain of the rat. PMID- 3037426 TI - Ca2+/calmodulin-dependent phosphoprotein phosphatase activity of calcineurin in rat striatum: effect of kainic acid lesions. AB - Calcineurin, a Ca2+ and calmodulin (CM)-dependent phosphatase, has been shown to be present in high concentrations in the striatum. Using inhibitor 1(phosphorylated by cAMP-dependent protein kinase) as a substrate, we found Ca2+/CM-dependent phosphatase (calcineurin) to be more than 2-fold higher than non-Ca2+/CM-dependent phosphatase in the rat striatum. In order to determine the cellular localization of calcineurin, striatal kainic acid injections were used to destroy neurons whose cell bodies are present at the site of injection. Glutamic acid decarboxylase (GAD) activity was measured as an indicator of destruction of striatal GABA-ergic neurons. After intrastriatal injection of 1 and 2 ug of kainic acid, there was a significant decrease of both calcineurin and GAD. However, injection of 0.5 ug kainic acid into the striatum reduced GAD activity by 81%, but had no effect on calcineurin phosphatase activity. Thus calcineurin does not appear to be equally distributed among all types of striatal neurons, but rather may be concentrated in neurons that are less sensitive to kainic acid than the GABA-ergic neuron. PMID- 3037428 TI - Tyr-D-Ala-Gly-(Me)Phe-chloromethyl ketone: a mu specific affinity label for the opioid receptor. AB - An alkylating tetrapeptide enkephalin derivative, Tyr-D-Ala-Gly-(Me)Phe chloromethyl ketone (DAMK) was synthesized, and its binding characteristics on rat brain membranes were evaluated. In competition experiments, the product shows high affinity for the mu opioid binding site of the rat brain membranes, whereas its binding to the delta and kappa subtypes is weak. Micromolar concentrations of this ligand produce a dose-dependent, apparently irreversible inhibition of /3H/ naloxone binding, with apparent IC50 value of 1-5 uM. Neither reversibly binding opioids nor tosyl-amino acid chloromethyl ketones show these effects. Saturation binding analysis with /3H/-naloxone of membranes preincubated with Tyr-D-Ala-Gly (Me)Phe-CH2Cl reveal a selective and irreversible inhibition of the high affinity /3H/-naloxone binding site. Irreversible blockade of mu-selective /3H/-ligand binding by Tyr-D-Ala-Gly-(Me)Phe-CH2Cl is much more effective than that of the binding of /3H/-enkephalin or /3H/-ethylketocyclazocine. The mu-selective binding properties of this new irreversible enkephalin analogue suggest that it could serve as an affinity label for the mu opioid receptor subtype. PMID- 3037427 TI - Intracerebroventricular administration of superFIT and its enantiomer to rats: evidence for in vivo acylation of [3H]DADL binding sites. AB - SuperFIT is an high affinity acylating ligand derived from fentanyl. Previous studies suggested that a selective acylation of delta receptors (J. Med. Chem. 29:1087-1093, 1986) resulted from exposure of membranes to this and structurally related compounds. We report in this preliminary study that intracerebroventricular administration of either superFIT or its enantiomer 18 to 24 hours prior to sacrifice decreased the subsequent binding of [3H]DADL to both its higher and lower affinity binding sites. PMID- 3037429 TI - Neuropeptide-Y acutely stimulates rat zona glomerulosa in vivo. AB - Neuropeptide-Y (NPY) acutely enhanced the plasma concentration of aldosterone (but not that of corticosterone) in both normal animals and in rats whose hypothalamo-hypophyseal axis and renin-angiotensin system were pharmacologically interrupted. The maximal response was obtained with a dose of 150 micrograms/kg. This dose of NPY raised the activity of 11 beta-hydroxylase and 18-hydroxylase in the capsular adrenal (zona glomerulosa), but not that of 11 beta-hydroxylase in the inner adrenocortical layers (zonae fasciculata and reticularis). These findings seem to indicate that NPY is specifically and directly involved in the acute stimulation of the late steps of the secretory activity of the rat zona glomerulosa. PMID- 3037430 TI - Metastases of a spinal glioblastoma multiforme into an intracranial arachnoid cyst. AB - Spread of spinal cord glioblastoma multiforme to the intracranial compartment is uncommon. This report documents a case of metastatic spread of glioblastoma multiforme from the spinal cord to an intracranial arachnoid cyst. Seeding of the cyst cavity could have occurred by direct permeation of the cyst wall by the neoplastic glial cells spreading via the cerebrospinal fluid or by direct access of the cells through a defect in the arachnoid cyst wall and their subsequent entrapment by a ball valve type mechanism, leading to multiple implants. PMID- 3037431 TI - Subarachnoid-pleural fistula complicating thoracotomy: case report and review of the literature. AB - We describe a patient who developed a persistent pleural effusion due to a subarachnoid-pleural fistula after operation for lung carcinoma and then review the literature on iatrogenic subarachnoid-pleural fistulas. PMID- 3037432 TI - Effect of hydrocortisone on the activity of the angiotensin-converting and renin like enzymes and kininase I in rat brain and hypophysis. PMID- 3037433 TI - Kainate binding sites in the hippocampal mossy fibers: localization and plasticity. AB - The regional distribution of high affinity binding sites for kainic acid has been determined in rat hippocampi by quantitative autoradiography. Selective lesions were made in order to determine the exact localization of these sites in the mossy fiber system, and to evaluate whether the sprouting and synaptic reorganization of the mossy fibers are associated with alterations in the distribution of these binding sites. The results show that kainate binding sites in the stratum lucidum are more vulnerable to destruction of the granules and their mossy fibers by intrahippocampal colchicine injections than to destruction of the CA3/CA4 pyramidal cells by injection of kainate into the amygdala. This suggests that a substantial proportion of the kainate binding sites is associated with the mossy fiber terminals (i.e. the presynaptic elements). Furthermore, in keeping with an earlier study, destruction of the pyramidal neurons of CA3 by intracerebral kainate produced a dark Timm positive band in the supragranular zone which is due to the sprouting of mossy fibers. This was associated with an increase in the density of kainate binding sites, which further stresses the parallelism between the distribution of these sites and mossy fiber terminals. PMID- 3037434 TI - Long-term blockade by toxin F of nicotinic synaptic potentials in cultured sympathetic neurons. AB - The effects of a recently identified blocker of neuronal nicotinic transmission, toxin F, were studied in cultured sympathetic neurons. Single principal neurons, dissociated from superior cervical ganglia of newborn rats, were grown on cardiac myocytes in microculture. The toxin blocked nicotinic synaptic potentials in these cultures but had no effect on muscarinic interactions. When toxin F was applied by addition to the perfusion medium, the concentration required for blocking most of the nicotinic potential was 40 nM, and the recovery from blockade was slow (t1/2 = 95 +/- 64 min). When the toxin was briefly applied by pressure ejection from a pipette, the concentration in the pipette necessary for blockade was 21 microM, and 30-60% of the response recovered within a few minutes while the remainder recovered slowly (t1/2 of the remainder = 105 +/- 82 min). One possible explanation for the difference in recovery time is that toxin F binds initially with low affinity to the nicotinic receptor but with time the toxin receptor complex converts to a high affinity state. The presence of dihydro beta-erythroidine during perfusion of toxin F prevented the long-lasting blockade by the toxin, suggesting that toxin F and dihydro-beta-erythroidine act through a common binding site. The specificity, potency, and slow reversibility of the effects of toxin F make it useful as a probe for studying neuronal nicotinic receptors of cultured sympathetic neurons. PMID- 3037435 TI - Analysis of quantal content and quantal conductance in two populations of neurons in the avian ciliary ganglion. AB - The avian ciliary ganglion contains two populations of parasympathetic cells, termed the ciliary and choroid neurons. We have estimated the quantal contents of nicotinic excitatory postsynaptic potentials in both populations of neurons by several methods. The singly innervated ciliary neurons have quantal contents of 15-30. In contrast, the multiply innervated choroid cells have quantal contents of 4-7. Quantal conductance was also determined, using a parallel conductance model which takes into account the capacitance of the cell membrane. This analysis indicates that in both populations of neurons one quantum activates approximately 100 postsynaptic receptors. It is concluded that in autonomic ganglia singly innervated cells demonstrate a larger quantal content, consistent with a higher safety factor for neurotransmission, while quantal content in multiply innervated cells is generally much lower, allowing for considerable summation of presynaptic inputs. Further, in autonomic neurons many fewer postsynaptic receptors are activated by a single quantum than is the case at the neuromuscular junction. PMID- 3037436 TI - Spontaneous quantal and subquantal transmitter release at the Torpedo nerve electroplaque junction. AB - Focal electrodes were used to record the spontaneous miniature potentials generated on delimited patches of innervated membrane in the Torpedo electric organ. The main population of miniature potentials followed a bell-shaped amplitude distribution. In addition, we observed a second class of spontaneous events that were smaller and whose amplitude distribution was skewed. These subminiatures formed an homogenous population together with the regular miniatures with respect to their time course versus amplitude relationship. They were thus probably generated at the same sites. The proportion of potentials that were subminiature was less than 10% in resting, freshly excised tissue, but it increased markedly: (i) when the tissue was kept for 24-28 h in vitro after excision; (ii) in the period following a brief heat challenger or (iii) stimulation to exhaustion; and (iv) in the presence of dinitrophenol or dinitrofluorobenzene. In all these conditions, we measured the acetylcholine, adenosine 5'-triphosphate and creatine phosphate content of the tissue and found a correlation between the relative number of subminiature potentials and the lack of energy rich molecules. It is concluded that subminiature potentials are present in the electric organ as in neuromuscular junctions. They are probably produced at the same sites as the regular miniature potentials and their relative occurrence seems to increase greatly when the nerve terminals are in a state of energy deficiency. PMID- 3037437 TI - Interferon treatment of experimental Ross River virus polymyositis. AB - Ross River virus (RRV), an alpha togavirus, causes an inflammatory myopathy in mice, which probably results from direct lytic effects of virus or viral products on myofibers. Administration of recombinant hybrid human leukocyte interferon alpha A/D (rIFN-alpha A/D) ameliorates clinical illness and reduces mortality from 86 to 42%. Peak concentrations of virus are reduced by 1,000-fold in serum and by 30-fold in muscle, but anti-RRV antibody production is not altered. Treatment with rIFN-alpha A/D dramatically reduces inflammation and necrosis in muscle. Beneficial effects of rIFN-alpha A/D on experimental, RRV-induced polymyositis result in part from inhibition of viral replication and spread, though immunomodulation might also play an important role. PMID- 3037438 TI - Delayed leukoencephalopathy in survivors with small cell lung cancer. AB - Six patients with small cell lung cancer developed a slowly progressive neurologic syndrome characterized by apathy, abulia, memory loss, gait ataxia, and corticospinal tract signs 26 to 50 months (mean, 35.2 months) after prophylactic cranial irradiation and systemic chemotherapy. In each case this was accompanied by CT and/or MRI evidence of changes in the periventricular white matter. These patients are long-term survivors (41 to 69 months) and do not have CNS metastases. PMID- 3037439 TI - Delayed neurotoxicity after ingestion of carbamate pesticide. AB - We studied a patient who ingested 27 gm (500 mg/kg) of carbaryl (1-naphthyl N methylcarbamate), a popular carbamate pesticide. After he recovered from acute cholinergic toxicity, acute weakness of arms and legs was accompanied by electrophysiologic findings consistent with axonal peripheral neuropathy. Recovery began at 1 week and continued for 9 months. A similar delayed neuropathy has been described with organophosphates but not with carbamate insecticides. PMID- 3037440 TI - Demyelinating neuropathy due to primary IgM kappa B cell lymphoma of peripheral nerve. AB - A 53-year-old man presented with a painful, demyelinating sensorimotor peripheral neuropathy with lymphomatous infiltration on sural nerve biopsy, but no evidence of systemic lymphoma. The neuropathy responded to cytotoxic therapy. Seven years later he developed generalized lymphadenopathy due to B cell lymphoplasmacytoid lymphoma, with a subpopulation of cells expressing a monoclonal pattern of IgM kappa. The lymphomatous infiltrate in the original nerve biopsy showed similar monoclonal IgM kappa reactivity. The mechanism of demyelination of the peripheral nerves may be similar to that described in patients with IgM kappa monoclonal gammopathies. PMID- 3037441 TI - Parkinson's disease and megacolon: concentric hyaline inclusions (Lewy bodies) in enteric ganglion cells. AB - Concentric hyaline inclusions (Lewy bodies), found in the cytoplasm of pigmented and nonpigmented neurons, are considered characteristic of idiopathic Parkinson's disease. The finding of cytoplasmic inclusions identical to Lewy bodies in ganglion cells of the colonic myenteric plexus in a patient with idiopathic Parkinson's disease and acquired megacolon suggests primary involvement of the enteric nervous system by Parkinson's disease. PMID- 3037442 TI - Sensory neuropathy as remote effect of cancer. PMID- 3037443 TI - [Anesthesiologic problems in transsphenoidal surgery of GH-secreting and ACTH secreting adenomas]. PMID- 3037444 TI - [Primary closure of the residual cavity after abdominoperineal amputation of the rectum]. PMID- 3037445 TI - [Retroperitoneal paraganglioma. Description of a clinical case]. PMID- 3037446 TI - [Involvement of the polypeptide system and ACE in Crohn disease]. AB - The polypeptidyc system was studied and serum concentrations of ACE (Angiotension Converting Enzyme) were assayed in patients with Crohn's disease in the active and stabilising phases. The study was undertaken on the assumption that as a granulomatous condition Crohn's disease involves the polypeptidyc system in much the same way as in sarcoidosis. The results confirm the hypothesis revealing a significant increase in ACE levels in patients with the active disease, whereas levels in the stabilized disease were lower than in the controls. It is suggested that, as in sarcoidosis, ACE levels are useful in the diagnosis, prognosis and follow-up of Crohn's disease, especially in relation to the use of steroid drugs. PMID- 3037447 TI - Cyclophosphamide in chronic progressive multiple sclerosis. PMID- 3037448 TI - Hypocalcaemic effect of WR-2721, S-2 (3-aminopropylamino) ethyl-phosphorothioic acid in an anuric haemodialysis patient. AB - The radio- and chemoprotective agent, S-2 (3-aminopropylamino) ethyl phosphorothioic acid (WR-2721) has been reported to lower hypercalcaemia in patients with cancer, probably by increased renal calcium excretion and decreased parathyroid hormone (PTH) secretion and bone calcium resorption. The present study reports the first clinical use of WR-2721 in an anuric haemodialysis patient with severe secondary hyperparathyroidism. The drug was administered intravenously at different doses, i.e. 150, 300, and 500 mg/m2. The infusion was followed by a striking decrease of plasma immunoreactive (i) PTH within 30 min. The nadir of the iPTH decrease was reached at 60 min and was followed by a steady return to previous values. Serum ionised calcium decreased more progressively from 1.55 mmol/l initially to 1.30 mmol/l at 4 h after the 300-mg dose, remained at that level at 24 h, but rose again to pre-infusion values after 48 h. The extent and duration of the decrease in plasma iPTH and ionised calcium were dose dependent. The circulating iPTH at 24 h was inversely related to the corresponding plasma ionised calcium concentration and had risen above preinfusion values at that time. Plasma concentrations of three other hormones, i.e. renin, insulin, and prolactin, were not affected by the administration of WR 2721. In conclusion, WR-2721 can induce a decrease in serum ionised calcium in the absence of any excretory kidney function. The rapid effect of the drug on circulating iPTH supports the notion of an interference with PTH secretion or catabolism. PMID- 3037449 TI - Evidence that neurokinin A (substance K) neurons project from the striatum to the substantia nigra in rats. AB - The striatonigral neurokinin A (NKA) pathway in rats was examined with an antiserum specific to the N-terminal region of NKA. Thermal lesions of the striatum decreased both substance P-like immunoreactivity (SP-LI) and NKA-like immunoreactivity (NKA-LI) in the substantia nigra (SN). The releases of SP-LI and NKA-LI in the SN were measured by a push pull superfusion technique. Addition of 50 mM K+ to the superfusion medium increased the releases of both SP-LI and NKA LI to 1.5-2.0 times the basal values. These results suggest that NKA neurons as well as SP neurons in the striatum project to the SN in rats. PMID- 3037450 TI - Dopamine receptors mediating the stimulation and the inhibition of adenylate cyclase in rat prostate gland. AB - The effect of dopamine on the 3',5'-cyclic adenosine monophosphate (cAMP) generating system of membrane particles of rat prostate gland was studied. Dopamine increased the concentration of cAMP in a dose-dependent manner. The selective D1 receptor inhibitor SCH 23390 caused a decrease in dopamine-elicited cAMP levels. Any effect of dopamine on prostatic cAMP concentration was abolished by the simultaneous addition to the incubation medium of SCH 23390 and of the D2 receptor blocking agent (-)-sulpiride. Also the D2 receptor agonist bromocriptine decreased cAMP levels. The present data indicate the existence, in the rat prostate gland, of two types of dopamine receptors mediating, respectively, the activation (D1 effect) and the inhibition (D2 effect) of adenylate cyclase activity. PMID- 3037451 TI - FMRF-NH2-like factor from neurohaemal organ modulates neuromuscular transmission in the locust. AB - YGGFMRFamide, FMRFamide and related peptides potentiate transmission at locust slow motor synapses. Since immunohistochemical evidence points to neurohaemal organs (NHO) as potential sources for endogenous RFamide-like peptides we have applied extracts from the metathoracic NHO, equivalent to 1/3 NHO, to the metathoracic extensor tibiae muscle. NHO-extract depolarizes the muscle fibre and increases its membrane resistance; enhances transmitter release; and increases the amplitude of contraction and the rate of relaxation. These effects are in good qualitative and quantitative agreement with those of YGGFMRFamide (5 X 10( 8) to 10(-7) M). Octopamine which partly acts like YGGFMRFamide cannot account for the NHO-effects. PMID- 3037452 TI - Advances in cancer: viruses as causes of human cancer. PMID- 3037453 TI - Dietary fiber and gastrointestinal function. PMID- 3037454 TI - Propionate and cholesterol homeostasis in animals. PMID- 3037455 TI - 201Tl per-rectal scintigraphy in primary hepatocellular carcinoma. AB - The results of 201Tl per-rectal scintigraphy in 10 patients with primary hepatocellular carcinoma (HCC) were presented together with the findings from contrast angiography, computed tomography and ultrasonography. 201Tl accumulation within the tumour was seen in seven of ten patients. This accumulation was thought to be due to 201Tl supply not from the portal vein but from the hepatic artery since significantly high heart to liver uptake (H/L) ratio from 0.71 to 1.21 and clear visualization of the heart and kidneys, indicating the presence of abundant portal-to-systemic shunting, were observed. Another three patients showed negative 201Tl accumulation within the tumour and near-normal H/L ratios from 0.32 to 0.47 which indicates little portal-to-systemic shunting. This finding reveals the evidence of the lack of 201Tl supply to the tumour from the portal vein. It seems that HCC does not receive any significant amount of blood flow from the portal system. PMID- 3037456 TI - Ligand-free, protein-bound technetium-99m iron-dextran enhancement of technetium pyrophosphate uptake in tumours. PMID- 3037457 TI - Trichloroacetic acid in the treatment of human papillomavirus infection of the cervix without associated dysplasia. AB - Human papillomavirus-induced cytologic changes were found in 379 (10.9%) of 3461 Papanicolaou smears done at the Gynecology Clinic at Wayne State University. Of these 379 patients, 98 (25.8%) had no cytologic evidence of dysplasia. Forty-six of the 98 patients, who had colposcopic and histologic evidence of human papillomavirus without dysplasia, agreed to participate in this study. All patients underwent a Q-Tip application of 85% trichloroacetic acid to the entire cervix, including the endocervical canal and the transformation zone, until the mucosa turned white. Coexistent vaginal and vulvar condyloma were treated simultaneously in six patients. The patients were reexamined at two-week intervals to ascertain the need for reapplication of trichloroacetic acid. No colposcopic evidence of human papillomavirus was found in 18 patients at the first visit, whereas 28 patients required reapplication of trichloroacetic acid. Three months after treatment, a follow-up Papanicolaou smear and colposcopic examination revealed no human papillomavirus infection in 31 patients. Seven patients had persistent infection and eight patients were lost to follow-up. Our preliminary data from this study suggest that 85% trichloroacetic acid is effective treatment of human papillomavirus infection of the cervix without dysplasia. PMID- 3037458 TI - Human papillomavirus associated with vaginal intraepithelial neoplasia in women exposed to diethylstilbestrol in utero. AB - Five out of 959 young women, exposed to diethylstilbestrol (DES) in utero, developed vaginal intraepithelial neoplasia while they were under follow-up in the Diethylstilbestrol-Adenosis Project at Baylor College of Medicine, Houston, Texas. We suggest that the development of the vaginal intraepithelial neoplasia at a younger age than usual may be caused by a higher susceptibility of the DES exposed patient to factors associated with the development of intraepithelial neoplasia. A common finding in all five women was the detection of the deoxyribonucleic acid (DNA) of human papillomavirus types 6 or 16 in their lesions, using high-stringency in situ hybridization. The role of human papillomavirus and herpes simplex virus in the etiology of intraepithelial neoplasia is discussed. Close follow-up is recommended for DES-exposed patients, especially those who have risk factors known to be associated with genital neoplasia. PMID- 3037459 TI - Quantitative deoxyribonucleic acid analysis of patients with mild cervical atypia: a potentially malignant lesion? AB - Quantitative deoxyribonucleic acid (DNA) analysis of cervical biopsy material from 32 women with cytologic, colposcopic, and histologic evidence of mild cervical atypia consistent with cervical intraepithelial neoplasia I, reactive atypia, or human papillomavirus infection was carried out using flow cytometry and Feulgen microspectrophotometry. Evidence of aneuploidy, ie, neoplastic transformation, was demonstrated in all cases of cervical intraepithelial neoplasia I and 68% of cases with human papillomavirus-induced atypia. These results support the growing impression that human papillomavirus-induced cervical atypia should be regarded as a true precursor of cervical neoplasia, and emphasize the necessity to refer patients with mild atypia for definitive diagnosis and management. PMID- 3037460 TI - [Complex treatment and prevention of recurrence in patients with ocular herpes]. PMID- 3037461 TI - Superoxide (O2-) assay monocyte adherence test for the detection of cell-mediated immunity in cancer patients. AB - The reactivity of mononuclear cells from patients with breast cancer to the 3 M KCl extract of breast tumor was tested using the superoxide (O2-) assay monocyte adherence test. The mononuclear cells were incubated with the 3 M KCl extract of breast tumor or normal breast tissues in a microcell for spectrophotometer analysis. After 2 h incubation, the nonadherent cells were removed, and the O2- generated from the monocytes adherent to the intersurface of the microcell was assayed. Thirty of 36 patients with breast cancer before treatment reacted to the extract of breast tumor. Sixteen patients showed adherence inhibition and 14 adherence stimulation. None of 8 patients with breast cancer reacted to the extract of lung tumor. Of 31 control donors (19 healthy volunteers and 12 patients with cancer of other organs), only 3 reacted to the extract of breast tumor. PMID- 3037462 TI - Immunohistochemical localization of phosphatidylinositol-4,5-bisphosphate in the rat lens. AB - We have attempted to localize immunohistochemically phosphatidylinositol-4,5 bisphosphate (PIP2) in rat lens tissue using affinity purified rabbit anti-PIP2 antibodies. Evidence indicates that PIP2 is localized to the lens epithelial cells but appears to be absent from the lens fiber cells. PMID- 3037463 TI - Changes of adrenergic and muscarinic cholinergic receptors in nasal mucosa in nasal allergy. AB - Receptor-binding assays were performed to evaluate the changes of beta- and alpha 1-adrenergic and muscarinic receptors in the nasal mucosa of subjects with nasal allergy and in guinea pigs sensitized with ovalbumin using radioligands 3H-DHA, 3H-prazosin and 3H-QNB, respectively. In subjects with nasal allergy, a decrease in density of beta- and alpha 1-adrenergic receptors and an increase in density of muscarinic cholinergic receptors were observed. An increase in density of muscarinic cholinergic receptors could be reproduced in the nasal mucosa of guinea pigs which were sensitized with ovalbumin and had typical hyperreactive nasal symptoms. These results indicate that the increase in the density of muscarinic receptors observed in the nasal mucosa of subjects with nasal allergy has been induced secondarily by an allergic reaction in the nasal mucosa with hyperreactive nasal symptoms, which in turn acts as an aggravating factor in the vicious circle promoting hyperreactivity of the nasal mucosa. PMID- 3037464 TI - Transfection of human nasopharyngeal epithelial cells with Epstein-Barr virus DNA. AB - Transfection experiments using Epstein-Barr virus (EBV) DNA were carried out to examine whether three different epithelial cell lines (Ad-AH, D98, and KB) could be transfected and whether biologically active virus could be recovered. We found that the Ad-AH cells derived from the human nasopharynx were more transfectable, as determined by the synthesis of EBV-specific antigens. Concentrates of supernatant fluids obtained from transfected Ad-AH cells were capable of transforming human cord blood lymphocytes (HCBLs). However, no concentrates of supernatant fluids obtained from transfected epithelial cell lines outside the nasopharynx (D98 and KB) were capable of transforming HCBLs. PMID- 3037465 TI - Effect of ACTH-(4-10) on Bechterew's compensation in squirrel monkeys. AB - ACTH-(4-10), a fragment of the ACTH without hormonal action, has been known to have effects on learning and memory process. Recently, we have shown that ACTH-(4 10) had marked effects on the balance compensation process after unilateral labyrinthectomy in squirrel monkeys. In this study, we examined the effect of daily administration of ACTH-(4-10) following two-staged bilateral labyrinthectomy. When the locomotor balance function and the slow-phase eye velocity of the spontaneous nystagmus were analyzed, the ACTH-(4-10) groups compensated significantly faster than the control group (p less than 0.05). PMID- 3037466 TI - [A rare case of adenoid cystic carcinoma of the larynx and trachea]. PMID- 3037467 TI - ANA reaffirms support for CDC guidelines. PMID- 3037468 TI - Increase of serum angiotensin-converting enzyme following venous stasis of the forearm in man. PMID- 3037469 TI - Continuing care of the spinal cord injured. AB - The functions of a comprehensive Spinal Cord Unit do not cease with the discharge of the patient from in-patient treatment after rehabilitation; they extend to aftercare, both in medical follow-up and prevention and treatment of complications, and in ongoing support and education of the patient and his family. An effective aftercare service must offer a life-long commitment to patients, and the aftercare team must include members of the disciplines which were involved in the initial rehabilitation, and also often others. Although demanding of time, resources and money, good aftercare is cost-effective in terms of savings in the cost of out-patient treatment and of re-admissions to hospital for the treatment of complications, and in maintaining many patients in the community, often as contributing members of society. PMID- 3037470 TI - The production of reactive oxygen species in mice infected or immunized with Plasmodium berghei. AB - The capacity of peritoneal exudate cells (PEC) obtained from mice infected or immunized with Plasmodium berghei to produce reactive oxygen species (ROS) and the biological basis of this response was investigated, using luminol-dependent zymosan-triggered chemiluminescence (CL). CL response of PEC from infected mice increased at the early stage but was significantly depressed at the later course of the infection. A similar biphasic activity of peroxidase was also observed in PEC from infected mice. On the other hand, PEC from immunized mice exhibited concomitant increases of the ability to produce CL, the activity of peroxidase and the expression of Fc and C3 receptors on cell surface. Compared with the controls, PEC from immunized mice showed an elevated background CL, responded more rapidly to the stimulation and generated considerably higher CL when triggered with opsonized zymosan. The data suggest that phagocytes in immunized mice are active in the production of ROS while those in infected mice are less active, and the inhibition of the ability of phagocytes to produce ROS may be one of the mechanisms for the parasites to escape from host immune system. PMID- 3037471 TI - Carcinoma of the colon: diagnosis by ultrasound and enema. AB - Carcinoma of the colon presenting before puberty is rare and few cases have been recorded in radiological literature. The symptomatology is usually vague but may be similar to the classical presentation of adults. The barium enema will also show the same constricting lesions. Two cases are reported, in one of which the initial diagnosis was made by ultrasound. PMID- 3037472 TI - Effect of group B streptococcal type-specific antigen on polymorphonuclear leukocyte function and polymorphonuclear leukocyte-endothelial cell interaction. AB - Neonatal group B streptococcal pneumonia is a severe disease, often resulting in death. Autopsy findings resemble those of hyaline membrane disease. Numerous organisms may be seen in the alveoli, but few polymorphonuclear leukocytes (PMNs) are found in the areas of bacterial invasion. Aggregated PMNs are often found, however, in the pulmonary capillaries. This study was designed to explore the effect of the group B streptococcal (GBS) type III antigen on PMN chemotaxis and PMN-endothelial cell interactions. Human PMNs were isolated and pretreated with 0.25 to 4 micrograms/ml of GBS type III antigen prior to determining their chemotactic response to the chemoattractants formyl-methionyl-leucyl phenylalanine, zymosan-activated serum, platelet-activating factor, and leukotriene B4. The GBS antigen caused a concentration-dependent inhibition of formyl-methionyl-leucyl-phenylalanine, zymosan-activated serum, and platelet activating factor-mediated chemotaxis (% inhibition of 38.1 +/- 4.0, 55.5 +/- 3.3, 46.7 +/- 9.7%, respectively; p less than 0.01). Leukotriene B4-mediated chemotaxis was not significantly depressed (21.2% +/- 7.7 inhibition; NSD). Group B streptococcal antigen also inhibited formyl-methionyl-leucyl-phenylalanine induced PMN adherence to endothelial cells in a concentration-dependent fashion when incubations were performed in the absence of serum. In contrast, incubation of GBS type III antigen with serum deficient in antibody to GBS resulted in a marked enhancement of PMN attachment to human endothelial cells. No significant enhancement of adherence was sen with the antigen in the presence of serum containing GBS type III antibody. These data suggest that the GBS type III antigen by itself may inhibit the influx of PMNs into the local site of infection in the alveoli.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3037473 TI - Neonatal adaptation: naloxone increases the catecholamine surge at birth. AB - A marked increase in plasma catecholamines at birth has been described in animals and man. Because the factors that regulate catecholamine secretion are incompletely understood and because it has recently been suggested that endogenous opiates are important in the regulation of catecholamine secretion, we designed studies to determine the influence of opiate receptor blockade prior to delivery on the increase in plasma catecholamines at birth. Term fetal sheep were delivered by cesarean section and randomly assigned to receive naloxone or vehicle. Naloxone was given just prior to umbilical cord cutting as a 2 mg/kg bolus followed by 2 mg/kg/h. Naloxone administration resulted in significantly greater peak levels of plasma norepinephrine (peak levels of 1.5 +/- 0.4 versus 0.9 +/- 0.1 ng/ml) and epinephrine (peak levels of 1.4 +/- 0.7 versus 0.9 +/- 0.3 ng/ml) and higher norepinephrine values throughout the study period. Naloxone administration was associated with significantly elevated heart rate (peak 184 +/ 12 versus 207 +/- 13 beats per min) and blood pressure (peak 95 +/- 6 versus 88 +/- 2 mm Hg). These studies demonstrate that opiate receptor blockade from birth markedly augments the neonatal sympathoadrenal response in the term newborn lamb. PMID- 3037474 TI - Varicella zoster immunoglobulin after postnatal exposure to varicella: survey of experts. AB - A questionnaire based on a hypothetical 6-day-old infant exposed to a sibling with varicella was mailed to 316 Pediatric Infectious Diseases Society members to determine their recommendations for administration of varicella zoster immunoglobulin (VZIG). Of the 137 faculty members who responded 20% would recommend VZIG for the infant described (13% were undecided). When further information including a negative maternal history for varicella, prematurity or maternal varicella infection occurring 1 week after delivery was considered, the numbers who recommended VZIG (43, 46 and 52%, respectively) rose significantly (P less than 0.01). Twenty-two physicians had knowledge of a severe case of varicella following postnatal exposure. Only 33% of faculty members surveyed feel there are no indications for VZIG in infants exposed after 48 hours of age. PMID- 3037475 TI - 1 alpha-hydroxyvitamin D3 treatment of three patients with 1,25-dihydroxyvitamin D-receptor-defect rickets and alopecia. AB - Three patients with clinically different severities of vitamin D-dependent rickets, type II, with alopecia, which is 1,25-dihydroxyvitamin D-receptor-defect rickets and is particularly resistant to treatment with calciferol analogues, were treated with large doses of 1 alpha-hydroxyvitamin D3 (1 alpha-(OH)D3) and 2 g of calcium lactate. Except for the alopecia, all of the abnormalities of patients 1 and 2 were reversed by treatment with 3 micrograms/kg/d of 1 alpha (OH)D3, and those of patient 3, who had the severest manifestations, were reversed by treatment with 6 micrograms/kg/d. The serum 24,25-dihydroxyvitamin D concentrations of the three patients were low before treatment and those of patients 1 and 2 increased during treatment. These findings suggest that in patients 1 and 2, 25-hydroxyvitamin D-24-hydroxylase was stimulated via a 1,25 dihydroxyvitamin D-receptor-mediated system by treatment with 1 alpha-(OH)D3. PMID- 3037476 TI - [Primary liver cancer accompanied by the circulation of the hepatitis B surface antigen in the blood serum of a 13-year-old child]. PMID- 3037477 TI - [Changes in the indices of cyclic nucleotides (cAMP and cGMP) in the blood plasma of children with acute intestinal infections]. PMID- 3037478 TI - Mucosal biopsy of the esophagus in children. PMID- 3037479 TI - Evidence for rheogenic sodium bicarbonate cotransport in the basolateral membrane of oxyntic cells of frog gastric fundus. AB - Ionic conductance properties of the basolateral cell membrane of oxyntic cells were studied in frog gastric fundus in vitro. After mounting the fundus in a modified Ussing chamber the serosal connective tissue was dissected off and individual oxyntic cells were punctured from the serosal surface with microelectrodes. Under resting conditions the membrane potential averaged -56.9, SD +/- 9.5 mV (n = 63), cytoplasm negative. Lowering or raising serosal HCO-3 concentration from 17.8 to 6 or 36 mmol/l respectively at constant PCO2 depolarized or hyperpolarized the cell membrane by +16.7 or -18.2 mV respectively. Sudden removal of serosal Na+ also depolarized the cell membrane (anomalous Nernst response). Since both the HCO-3 dependent and the Na+ dependent potential changes were strongly depressed by the disulfonic stilbene SITS and since the potential response to HCO-3 was virtually abolished in Na+-free solution we conclude that a rheogenic Na+ (HCO-3)n-cotransport system (n greater than 1) is present in the basolateral cell membrane of oxyntic cells. Its possible role in base transfer during HCl-secretion or HCO-3 secretion remains to be elucidated. PMID- 3037480 TI - Effect of triiodothyronine on system A amino acid transport in cells of rat submandibular gland. AB - The effect of L-3,5,3'-triiodothyronine (T3) on alpha-aminoisobutyric acid (AIB) transport in isolated cell suspensions of rat submandibular gland was investigated. The uptake of ATB by these cells appeared to require extracellular Na+ and was inhibited by ouabain (10(-3) M). Cell suspensions from thyroidectomized rats which have been given three successive doses of T3 on alternate days (50 micrograms/100 g BW) showed a significant increase in AIB uptake compared with cells isolated from thyroidectomized rats treated with diluent. Efflux of AIB from the cell suspension was not affected by T3. There was no significant changes in AIB uptake 12 h after a single injection of T3 (50 micrograms/100 g BW). However, there was a significant 49% and 65% increase in AIB net uptake at 24 and 48 h, respectively, after T3 treatment. Under similar conditions, the cell suspension showed a 48% increase in NaK-ATPase activity at 12 h and to a peak of 61% at 24 h. Therefore, changes in NaK-ATPase activity preceded the changes in AIB net uptake upon treatment with T3, implying that AIB uptake is probably mediated, at least in part, by the activity of NaK-ATPase. PMID- 3037481 TI - Mechanisms regulating the adrenaline-induced long-term potentiation in bullfrog sympathetic ganglia. AB - Two regulatory mechanisms on the long-term potentiation of transmitter release induced by adrenaline (adr.-l.t.p.) in bullfrog sympathetic ganglia were studied by recording intracellularly the fast excitatory postsynaptic potentials. An increase in exposure time to adrenaline from 10 min to 60 min did not enhance the magnitude of adr.-l.t.p. However, increasing an exposure time to dibutyryl cyclic AMP (1 mM) up to 60 min progressively enhanced the magnitude of the nucleotide induced potentiation, indicating the desensitization of the beta-adrenoceptor. The desensitization remained at 20 min after the removal of adrenaline in all the five cells but disappeared at 60-90 min in four cells out of eight. Under the latter condition, the second l.t.p. was summated on the first one. Dibutyryl cyclic GMP (100 microM) blocked the generation of the l.t.p. induced by dibutyryl cyclic AMP (1 mM) as well as that of adr.-l.t.p. Muscarine (10 microM) or adenosine (1 mM), a possible candidate for raising intraterminal cyclic GMP, did not significantly affect adr.-l.t.p. These results suggest that adr.-l.t.p. is regulated by the desensitization of beta-adrenoceptor and a process which involves endogenous cyclic GMP acting on a step subsequent to the cyclic AMP production. PMID- 3037482 TI - Activation of luminal Na+/H+ exchange in distal nephron of frog kidney. An early response to aldosterone. AB - Increased chronic intake of K+ induces H+ and K+ secretion in amphibian distal tubule, paralleled by an elevation of plasma aldosterone. The present experiments test whether the mineralocorticoid hormone is responsible for the alteration of ion transport. The blood capillaries of the isolated kidneys of NaCl-adapted (i.e. aldosterone-suppressed) Rana pipiens were perfused with HEPES-buffered amphibian Ringer solution (pH 7.8). Limiting intraluminal pH (pHlu) was measured continuously with pH-sensitive microelectrodes while aldosterone (3 X 10(-7) to 3 X 10(-6) mol/l) was applied in the peritubular perfusate. Concomitant with a decrease of the lumen-positive transepithelial potential (Vte) from 8.5 +/- 1.1 mV to 4.0 +/- 0.6 mV pHlu dropped from 7.73 +/- 0.02 to a new steady-state value of 7.17 +/- 0.05 within 60 to 180 min of aldosterone administration. Significant luminal acidification occurred already 20 min after application of aldosterone. Luminal addition of 10(-3) mol/l amiloride reversed luminal acidification to a pHlu of 7.68 +/- 0.04; at the same time Vte recovered partially. Pretreatment of the distal tubules with spironolactone prevented the aldosterone-induced acidification of the tubule fluid. We conclude that in early distal tubule of the amphibian kidney aldosterone--after interaction with cytoplasmic receptors- activates the luminal, amiloride-inhibitable Na+/H+ exchanger. This mechanism could explain enhanced H+ secretion found in the K+ adapted animal. PMID- 3037483 TI - Enzymatic amplification of translation inhibition of rabbit beta-globin mRNA mediated by anti-messenger oligodeoxynucleotides covalently linked to intercalating agents. AB - The effects of anti-messenger oligodeoxynucleotides, covalently linked to an intercalating agent, on translation of rabbit beta-globin mRNA, were investigated both in wheat germ extract and in microinjected Xenopus oocytes. A specific inhibition of beta-globin synthesis was observed in both expression systems with a modified 11-mer covalently linked to an acridine derivative. In injected oocytes a more efficient block was observed with this modified oligonucleotide than with its unsubstituted homolog. This was ascribed to stacking interactions of the intercalating agent with base pairs which provide an additional stabilization of the [mRNA/DNA] hybrid. We demonstrated that in wheat germ extract, the modified and unmodified oligonucleotides behaved similarly due to the presence of a high RNaseH activity. RNaseH was also present, although to a lesser extent, in the oocyte cytoplasm. This anti-messenger DNA-induced degradation of target mRNA resulted in amplified efficiency of hybrid-arrested translation. This additional mechanism might provide anti-sense DNAs with an advantage over anti-sense RNAs. PMID- 3037484 TI - c-myc protein can be substituted for SV40 T antigen in SV40 DNA replication. AB - Replicating activity of SV40 origin-containing plasmid was tested in human cells as well as in monkey CosI cells. All the plasmids possessing SV40 ori sequences could replicate, even in the absence of SV40 T antigen, in human HL-60 and Raji cells which are expressing c-myc gene at high level. The copy numbers of the replicated plasmids in these human cells were 1/100 as high as in monkey CosI cells which express SV40 T antigen constitutively. Exactly the same plasmids as the transfected original ones were recovered from the Hirt supernatant of the transfected HL-60 cells. Furthermore, replication of the SV40 ori-containing plasmids in HL-60 cells was inhibited by anti-c-myc antibody co-transfected into the cells. These results indicate that the c-myc protein can be substituted for SV40 T antigen in SV40 DNA replication. PMID- 3037485 TI - Evidence for the involvement of the 16kD gene promoter in initiation of chromosomal replication of Escherichia coli strains carrying a B/r-derived replication origin. AB - Initiation of chromosomal DNA replication of several Escherichia coli dnaA (Ts) strains is diminished in cell harbouring pBR322 hybrid plasmids carrying both oriC and the adjacent 16kD gene promoter of E. coli K12. This perturbance, resulting in very slow growth, is caused both by the dnaA allele and the E. coli B/r-derived region of the replication origin of these strains. Cloning and DNA sequence analysis of the E. coli B/r replication origin revealed several base differences as compared to the E. coli K12 sequence. The replication origin of temperature sensitive fast growing mutants, originating from a homologous exchange between chromosomal and plasmid DNA sequences were also cloned. Sequence data showed that a single base change within the promoter of the 16kD gene of these dnaA (Ts) strains is able to suppress the inhibition of chromosomal DNA replication by the mentioned pBR322 hybrid plasmids. Our results strongly indicate a role of the 16kD gene promoter in control of initiation of chromosomal DNA replication. PMID- 3037486 TI - Nucleotide sequence and LexA regulation of the Escherichia coli recN gene. AB - The nucleotide sequence of a 2224 bp region of the Escherichia coli chromosome that carries the LexA regulated recN gene has been determined. A region of 1701 nucleotides encoding a polypeptide of 567 amino acids with a predicted molecular weight of 63,599 was identified as the most probable sequence for the recN structural gene. The proposed initiation codon is preceded by a reasonable Shine Dalgarno sequence and a promoter region containing two 16 bp sequences, separated by 6 bp, that match the consensus sequence (SOS box) for binding LexA protein. DNA fragments containing this putative promoter region are shown to bind LexA in vitro and to have LexA-regulated promoter activity in vivo. The amino acid sequence of RecN predicted from the DNA contains a region that is homologous to highly conserved sequences found in several DNA repair enzymes and other proteins that bind ATP. A sequence of 9 amino acids was found to be homologous to a region of the RecA protein of E. coli postulated to have a role in DNA/nucleotide binding. PMID- 3037487 TI - Supercoiling in prokaryotic and eukaryotic DNA: changes in response to topological perturbation of plasmids in E. coli and SV40 in vitro, in nuclei and in CV-1 cells. AB - Changes in DNA linking number have been observed in plasmid DNA purified from E. coli cells after the cells were treated with chloroquine. Chloroquine, a DNA intercalating drug, unwinds the DNA, decreasing the levels of negative supercoiling. Following this in vivo topological perturbation, within minutes DNA gyrase decreases DNA linking number producing more negatively supercoiled DNA topoisomers. Following the removal of the drug from cells, within minutes topoisomerase 1 or DNA gyrase increases the linking number restoring the original level of supercoiling. Analogous changes in DNA linking number after addition of chloroquine are observed in purified plasmid DNA, and in purified SV40 minichromosomes in the presence of exogenous topoisomerase. Changes in linking number are also observed in SV40 chromosomes in isolated nuclei and in SV40 DNA purified from CV-1 cells following topological perturbation with chloroquine. These results suggest that eukaryotic cells may have mechanisms to maintain a defined level of DNA supercoiling. PMID- 3037488 TI - Cloning and mapping of infA, the gene for protein synthesis initiation factor IF1. AB - The gene for translation initiation factor IF1, infA, has been identified by using two synthetic oligonucleotides to screen a Charon 30 library of Escherichia coli DNA. A recombinant lambda phage, C1921, was purified from a plaque positive for both probes. A 2.8 kb BglII fragment and a 2.0 kb HindIII fragment isolated from C1921 were subcloned into the BamHI and HindIII sites of pBR322 to yield pTB7 and pTH2 respectively. Synthesis of IF1 in maxicells transformed with pTB7 or pTH2 indicates the presence of inf A in both inserts. This was confirmed by DNA sequencing: a region was found that codes for a 8,119 dalton protein with an amino acid sequence corresponding to IF1. The chromosomal location of inf A was determined by mapping the closely linked beta-lactamase gene (Ampr) in pTB7 and pTH2. pTB7 and pTH2 were transformed into polA Hfr hosts, and integration of the plasmid by homologous recombination near inf A was selected on the basis of ampicillin resistance. The site of integration was confirmed by Southern blot analysis of restriction nuclease digested wild type and transformed genomic DNA. The Ampr marker (and therefore inf A) was mapped to about 20 minutes by Hfr interrupted matings and P1 transduction experiments. The structure and regulation of the inf A operon currently are being investigated. PMID- 3037489 TI - Structure and expression of a human gene coding for a 71 kd heat shock 'cognate' protein. AB - In all eukaryotes examined so far, hsp70 gene families include cognate genes (hsc70) encoding proteins of about 70 Kd which are expressed constitutively during normal growth and development. We have investigated the structural relationship of heat-inducible and cognate members of the human hsp70 gene family. Among several human genomic clones isolated using Drosophila hsp/hsc70 probes, one contained an hsc70 gene. Its complete sequence is reported here. It is split by eight introns and encodes a predicted protein of 70899 d that would be 81% homologous to hsp70. Structural comparisons with corresponding genes from other species provide one of the most striking examples of gene conservation. Isolation of a corresponding cDNA clone, RNA-mapping and in vitro translation data demonstrate that the gene is expressed constitutively and directs the synthesis of a 71 kd protein. The latter is very likely to be identical to a clathrin uncoating ATPase recently identified as a member of the hsp70-like protein family. PMID- 3037490 TI - Transcription of a human transposon-like sequence is usually directed by other promoters. AB - The transcriptional activity of a human transposon-like family of repeats, called the THE-1 family, has been studied in cell culture and in human tissue. Both strands of THE-1 are present in several discrete length poly A plus RNAs. Primer extension studies and the structures of cDNA clones show that these THE-1 transcripts are usually the product of other transcription units. The THE-1 LTR provides the polyadenylation processing site for two transcripts, which result from upstream non THE-1 promoters. Yet another transcript, containing an internal THE-1 element in the probable sense orientation, is greatly enriched in a polysomal size fraction. PMID- 3037491 TI - In vivo transcription from multiple spacer rRNA gene promoters during early development and evolution of the intergenic spacer in the brine shrimp Artemia. AB - The control of ribosomal RNA (rRNA) gene expression during development can be productively studied by examination of the relationship between promoter structure and function as well as the processing of primary transcripts. Toward this end total cell RNA was extracted from embryos at various stages and probed with cloned rRNA genes using the "dot blot" method. This exercise showed that rRNA gene expression is a stage-specific process and is thus under developmental control. S1 nuclease protection experiments localized fourteen different upstream DNA sites encoding 5'-termini of pre-rRNAs during this synthetic phase of development. There is no indication of any spacer fail-safe terminator function. The S1 approach contributed to the sequencing of several of the sites. Comparative sequence alignments reveal short conserved regions in DNAs corresponding to these sites, which are shown to fall into two structural classes. Sites 3, 4, 6 and 9 are proposed to function in transcription initiation and are found to have the consensus sequence 5'...T-A-T-A-T-Pu-Pu-Pu-G-Pu-Pu-G-T C-A 3'. Sites 1, 2, 5 and 8 which are proposed to function in 5'-processing have the consensus sequence; 5'...Pu-G-T-Pu-T-T-G 3'. These short sequence conserved regions are hypothesized to serve as recognition signals for proteins within the rDNA transcription initiation complex and for 5'-processing enzymes, respectively. Sequencing of the intergenic spacer region from which a model for spacer evolution is derived shows that tandem ca 600 bp subrepeats explain much of the multiplicity observed within control sites. PMID- 3037492 TI - The SV40 termination region exhibits an altered helical DNA conformation. AB - The DNA structure of a fragment containing the SV40 termination sequences was examined using gel mobility assays. The region is shown to contain a DNA bend as evidenced by an abnormal mobility that is progressively accentuated as the temperature is lowered. This represents the strongest example of DNA bending among the collection of SV40 fragments studied. The same fragment was shown previously to uniquely support hyper-stable nucleosome formation in vitro, suggesting a possible relationship between DNA bending and nucleosome stability. PMID- 3037494 TI - pKM101 is an IS46-promoted deletion of R46. PMID- 3037493 TI - Human cardiac myosin heavy chain genes and their linkage in the genome. AB - Human myosin heavy chains are encoded by a multigene family consisting of at least 10 members. A gene-specific oligonucleotide has been used to isolate the human beta myosin heavy chain gene from a group of twelve nonoverlapping genomic clones. We have shown that this gene (which is expressed in both cardiac and skeletal muscle) is located 3.6kb upstream of the alpha cardiac myosin gene. We find that DNA sequences located upstream of rat and human alpha cardiac myosin heavy chain genes are very homologous over a 300bp region. Analogous regions of two other myosin genes expressed in different muscles (cardiac and skeletal) show no such homology to each other. While a human skeletal muscle myosin heavy chain gene cluster is located on chromosome 17, we show that the beta and alpha human cardiac myosin heavy chain genes are located on chromosome 14. PMID- 3037495 TI - pPyLT1 does not always dictate the formation of autonomously replicating elements in transgenic mice. PMID- 3037496 TI - cDNA encoding a human homolog of yeast ubiquitin 1. PMID- 3037497 TI - CAT constructions with multiple unique restriction sites for the functional analysis of eukaryotic promoters and regulatory elements. PMID- 3037498 TI - Human metallothionein-specific riboprobes. PMID- 3037499 TI - The cleavage site for the restriction endonucleases BanI and HgiC I is 5' ...G decreases GPyPuCC ...3'. PMID- 3037500 TI - [Evaluation of the effectiveness of surgical treatment of breast sarcomas developing as cystosarcoma phyllodes]. PMID- 3037501 TI - In case of AIDS. PMID- 3037502 TI - The journal of infection control nursing. AIDS Q&A. PMID- 3037503 TI - [Advances in studies on new converting enzyme inhibitors]. PMID- 3037504 TI - [Effect of treatment with pindolol, acebutolol and propranolol on the density of beta-adrenergic receptors in human lymphocytes]. PMID- 3037505 TI - Epithelial expression of HLA-DR antigens in laryngeal papillomas with a dense inflammatory reaction. AB - Thirty six laryngeal papillomas taken from 11 children and 10 adults were investigated with an indirect immunofluorescence technique for the occurrence of T cell subsets, Langerhans cells and HLA-DR antigen using specific monoclonal antibodies. A mild local cellular immune response was preferentially associated with juvenile laryngeal papillomas, whereas a more intense inflammatory reaction with T8 cell infiltration and large amounts of Langerhans cells in the chorion and the epithelium was observed in adult lesions. Some lesions either from adults or children characterized by high densities of lymphoid and dendritic cells exhibited the expression of DR antigen by epithelial cells. This result may suggest either a role of lymphokines released by activated T cells or a direct effect of viral infection on epithelial cells inducing DR antigen in laryngeal papillomas. PMID- 3037506 TI - Undifferentiated tumours of salivary glands. Immunocytochemical investigations and differential diagnosis of 22 cases. AB - 22 undifferentiated tumours of the Salivary Gland Register (University of Hamburg) were studied by conventional light microscopical and immunocytochemical methods to elucidate the heterogeneity of this tumour group. The following observations were made in this collective: 18 tumours displayed one or more markers for the epithelial character and were classified as carcinomas. 10 carcinomas were considered as primary ones and 8 were considered as secondary ones (metastatic or invasive "per continuitatem"). Primary carcinomas were subclassified as poorly differentiated variants of a distinctive type of salivary gland tumours, as follows: 6 cases of carcinoma in pleomorphic adenoma, and one case each of mucoepidermoid tumour, adenocarcinoma, salivary duct carcinoma and epidermoid carcinoma. Secondary carcinomas were subclassified as follows: 3 epidermoid carcinomas, 3 nasopharyngeal carcinomas and 2 bronchial carcinomas. One tumour positive for S-100 protein and NSE (Neuron-specific enolase) was classified as a metastatic melanoma. Another tumour positive for vimentin and actin was classified as a rhabdomyosarcoma of the periglandular tissue. Two tumours lacked any markers studied here and were regarded as a malignant paraganglioma and an undifferentiated lymphoma, respectively. The differential diagnosis of the undifferentiated tumours of salivary glands and the special problems of this tumour group are discussed. PMID- 3037507 TI - Size dependent enzyme activities of multinucleated (osteoclastic) giant cells in bone tumors. AB - In osteoclastic giant cells of six different tumors of bones and joints (fibrous dysplasia, proliferating giant cell tumor, malignant giant cell tumor, osteosarcoma after chemotherapy, malignant synovioma and Ewing's sarcoma) activities of tartrate-resistant acid phosphatase, NADH-tetrazolium oxidoreductase and, in three of them, of non-specific esterase are determined by enzyme histochemical methods. Quantitative microphotometry makes it possible to determine relative enzyme activities in the cut sections of giant cells of different sizes. Giant cells of the various tumors reveal similar trends: With an increase in cell size, mean extinctions of NADH-tetrazolium-oxidoreductase and non-specific esterase decrease. Mean extinctions of tartrate-resistant acid phosphatase increase in cells of medium size, whereas the large cells reveal in part low activities. An additional ultrastructural examination of the giant cells in the proliferating giant cell tumor as well as in the osteosarcoma shows morphological signs of degeneration in the large cells. Electron probe microanalysis of the proliferating giant cell tumor exhibits evidence of phagocytosis of Ca and/or Fe containing particles. The similar size dependent reaction pattern of enzymes in osteoclastic giant cells of different tumors favors the concept of a common histogenesis, i.e. a host reaction. PMID- 3037508 TI - Key issues in nutrition. Disease prevention through adulthood and old age. AB - Certain dietary practices are valid methods of lowering the risk of disease. Others, while popular, have unproven benefits or may even be associated with risks of their own. Careful evaluation of hypercholesterolemia is necessary. Persons with a high level of low-density lipoprotein (LDL) cholesterol and a low level of high-density lipoprotein (HDL) cholesterol need diet therapy, because they are at increased risk of cardiovascular disease. Weight reduction and fat restriction can lower blood pressure, help control hyperglycemia, and improve the LDL cholesterol-HDL cholesterol ratio. Some evidence indicates a protective role of beta carotene against cancer in animals. However, hypervitaminosis A is dangerous and relatively easy to accomplish, so supplementation beyond a multivitamin tablet is discouraged. Data about inhibition of cancer in humans through use of high doses of vitamin E or C or selenium are inconclusive, and studies of effects of long-term ingestion are not available. In general, megadoses of even healthy substances are thought to be dangerous. Decreased consumption of fat, increased consumption of foods high in fiber, and elimination of alcohol and tobacco are sensible recommendations. Consumption of cruciferous vegetables has not been proven to reduce the incidence of cancer, but a moderate amount of them in the diet would seem reasonable. PMID- 3037509 TI - The role of beta-glucan receptors on blood and tissue leukocytes in phagocytosis and metabolic activation. PMID- 3037510 TI - High performance purification of glycolytic enzymes and creatine kinase from chicken breast muscle and preparation of their specific immunological probes. AB - We developed a novel procedure for isolation of the muscle isozymes of aldolase, triose phosphate isomerase (TPI), glyceraldehyde phosphate dehydrogenase (GAPDH), phosphoglycerate kinase (PGK), phosphoglycerate mutase (PGM), enolase, pyruvate kinase (PK) and lactic dehydrogenase (LDH), and also creatine kinase (CK), at high purity, specific activity and yield. Protein was extracted from chicken breast muscle and glycolytic enzymes were purified by a three step procedure consisting of: Ammonium sulfate combined with pH fractionation. Phosphocellulose chromatography with performance of high pressure liquid chromatography, exploiting a pH gradient formed by a gradient of the buffering ion for protein elution. Affinity chromatography causing elution by substrate or pH. The enzymes, obtained at over 95% purity as judged by specific activity and silver stained electropherograms, were injected into sheep. Antibody for each enzyme was purified on specific immunosorbant and its specificity was verified by immunotransfer analysis. PMID- 3037511 TI - Purification and characterization of palmitoyl-CoA ligase from rat brain microsomes. AB - We have previously shown the existence of two separate enzymes for the synthesis of palmitoyl-CoA and lignoceroyl-CoA in rat brain microsomal membranes (1). Palmitoyl-CoA ligase activity was solubilized from brain microsomal membranes with 0.3% Triton X-100 and purified 93-fold by a combination of protein purification techniques. The Km values for the substrates palmitic acid, CoASH and ATP were 11.7 microM, 5.88 microM and 3.77 mM respectively. During activation of palmitic acid ATP is hydrolyzed to AMP and pyrophosphate, as evidenced by the inhibition of this activation by 5 mM concentrations of AMP, pyrophosphate or AMP and pyrophosphate to 70%, 60% and 85% respectively. The divalent metal ion Mg2+ was required for activity; its replacement with Mn2+ resulted in a 35% decrease in activity. Palmitoyl-CoA ligase activity was inhibited by the addition of oleic or stearic acids whereas addition of lignoceric acid or behenic acid had no effect. This supports our previous observation that palmitoyl-CoA and lignoceroyl CoA are synthesized by two different enzymes in rat brain microsomal membranes. PMID- 3037512 TI - [Peripheral neuropathy and HIV retrovirus infection]. PMID- 3037513 TI - [Malignant fibrous histiocytoma. Post-irradiation complication of Hodgkin's disease?]. PMID- 3037514 TI - [Influence of excitatory amino acids on the outcome of cerebral ischemia]. AB - It would appear from an analysis of the molecular mechanisms which determine the post-ischaemic destruction of brain tissue that they primarily involve the action of excitatory aminoacids, together with the inhibition of protein synthesis and the disorderly production of free radicals. The activities of endogenous excitatory aminoacids (glutamate, aspartate) immediately after ischaemia are similar to those of exogenous excitotoxins (kainic acid, N-methyl-D-aspartate). Brain tissue destruction consecutive to ischaemia or hypoglycaemia is mainly found in cerebral structures which contain a large number of glutamate receptors (hippocampus, locus coeruleus, corpus striatum). These lesions are associated with a significant increase in extracellular concentrations of excitatory aminoacids which are responsible for the massive penetration and accumulation of calcium in the cells (nucleus, Golgi complex, mitochondria), and this encourages the synthesis of non-functional proteins and/or proteolysis (direct cytotoxic action). In animals, these post-ischaemic necrotic lesions are prevented by administration of pharmacological antagonists of these neuromediators at receptor level (e.g. 2-amino-7-phosphonoheptanoic acid). These experimental results suggest that excitatory aminoacids play a determinant role in the genesis of post ischaemic cell death and that the glutamatergic antagonists currently under development may have a therapeutic activity. PMID- 3037515 TI - [Cerebral edema: intra-cellular or extra-cellular mechanisms?]. AB - Many cerebral pathological processes are attended by edema defined as an increase in brain volume associated with an increase in brain water and sodium contents. The aggravation of lesions induced by this edema warrants a pharmacological and therapeutic approach based on a detailed knowledge of its physiopathological mechanisms. Experimental models and in vitro studies have shown that the fundamental mechanisms leading to edema are: cold, acute hypoxia, ischaemia, arachidonic acid, toxic substances and plasma hypo-osmolarity. To the various types of edema described (vasogenic, cytotoxic, hydrocephalic) correspond different mechanisms. Vasogenic edema essentially depends on osmotic and hydrodynamic factors; cytotoxic edema results from perturbations in energy dependent cellular osmoregulation. The underlying biochemical disorders have now been demonstrated, mostly in ischaemic edema; they include, during the revascularization phase, disruption of the blood-brain barrier (vasogenic component) and changes in ion concentrations, neurotransmitters and energetic mechanisms. Key factors in the development of edema are cyclic AMP, serotonin and Na-K-ATPase. PMID- 3037516 TI - [Reduction of hospital costs in digestive surgery after treatment with immunostimulants]. PMID- 3037517 TI - Lentiviruses/HTLV-III. PMID- 3037518 TI - Origins of netropsin binding affinity and specificity: correlations of thermodynamic and structural data. AB - We report complete thermodynamic profiles for netropsin binding to an oligomeric and to several polymeric DNA host duplexes. These data allow us to reach the following conclusions: netropsin binding by deep penetration into the minor groove is overwhelmingly enthalpy driven and exhibits a very high binding affinity (K approximately 10(9) at 25 degrees C); deep penetration into the minor groove is required to form those drug-DNA interactions responsible for the enthalpy-driven high binding affinity of netropsin; I-C base pairs form binding sites for netropsin that thermodynamically are equivalent to those formed by A-T base pairs; the positive binding entropies reflect entropic contributions from molecular events other than just water spine disruption; the thermodynamic binding data primarily reflect local netropsin-DNA interactions rather than long range binding-induced conformational changes at regions distant from the binding site; the enhanced binding affinity associated with deep penetration of netropsin into the minor groove does not result from more favorable electrostatic interactions; the binding of netropsin to the central AATT core of the decamer duplex [d(GCGAATTCGC)]2 is thermodynamically modeled best by netropsin binding to the poly[d(AT)].poly[d(AT)] duplex rather than the poly(dA).poly(dT) duplex. We propose correlations between our thermodynamic data and specific molecular interactions defined by NMR and x-ray structural studies on similar and identical drug-DNA complexes. PMID- 3037519 TI - Proofreading by DNA polymerase III of Escherichia coli depends on cooperative interaction of the polymerase and exonuclease subunits. AB - The polymerase subunit (alpha) of Escherichia coli DNA polymerase III holoenzyme and the 3'----5' exonuclease subunit (epsilon) are each less active separately than together in the holoenzyme core (an assembly of alpha, epsilon, and theta subunits). In a complex formed from purified alpha and epsilon subunits, polymerase activity increased 2-fold, and that of the 3'----5' exonuclease increased 10- to 80-fold. The alpha-epsilon complex contains one each of the subunits as does the core. Stimulation of 3'----5' exonuclease activity is due mainly to a greatly increased affinity of the epsilon subunit for the 3'-hydroxyl terminus, resulting from DNA binding by the alpha subunit. Proofreading in the course of DNA synthesis by the alpha-epsilon complex was indistinguishable from that of the core. These findings identify the participation of the alpha subunit in proofreading by polymerase III holoenzyme and suggest that the fidelity of DNA replication may be influenced by the relative levels of the alpha and epsilon subunits in the cell. PMID- 3037520 TI - The gluconate operon gnt of Bacillus subtilis encodes its own transcriptional negative regulator. AB - The gluconate (gnt) operon of Bacillus subtilis consists of four gnt genes; the second and third genes code for gluconate kinase (gluconokinase, EC 2.7.1.12) and gluconate permease, respectively. A fragment carrying the promoter of this operon (gnt promoter) and the first gene (gntR) was subcloned into a promoter probe vector (pPL603B). Repression of the expression of cat-86 gene, encoded in the vector portion of a constructed plasmid (pgnt21), that is under the control of the gnt promoter was removed by gluconate. The results of deletion analysis and of insertional inactivation of the gntR gene cloned in pgnt21 suggested that the product of the gntR gene, actually synthesized as a 29-kDa protein in vivo, is involved in repression of the gnt promoter. A 4-base-pair insertional mutation within the gntR gene constructed in vitro was introduced into the B. subtilis chromosomal gnt operon by use of linkage of the 4 base pairs to gntK10 in transformation. The introduced mutation gntR1 caused the constitutive expression of the gluconate kinase and gluconate permease genes. S1 nuclease analysis indicated that the mRNA of this operon is synthesized in the gntR1 strain and amounts of mRNA are not changed very much by gluconate, which acts as an inducer in the wild-type gene. These results strongly indicate that the gntR gene codes for a transcriptional negative regulator for the gnt operon. PMID- 3037521 TI - Down-regulation of class I HLA antigens and of the Epstein-Barr virus-encoded latent membrane protein in Burkitt lymphoma lines. AB - Epstein-Barr virus (EBV)-carrying Burkitt lymphoma (BL) cells are relatively or completely resistant to the lytic effect of major histocompatibility complex class I HLA antigen-restricted cytotoxic T lymphocytes (CTLs) generated by stimulating lymphocytes of EBV-seropositive donors with the autologous EBV transformed lymphoblastoid cell line (LCL). We previously found that EBV-negative and EBV-carrying BL lines derived from HLA-A11-positive donors were not only resistant to lysis by the HLA-A11-restricted CTL generated by stimulation with the autologous LCL, but also to HLA-A11-specific CTL derived from lymphocytes of an EBV-seronegative donor stimulated with an allogeneic LCL. Using the same and additional cell lines, we now show that the CTL resistance of the BL lines is probably due to a selective down-regulation of HLA-A11. We also show that the EBV encoded latent membrane protein is expressed at a lower level in the EBV-carrying BL lines than in EBV-transformed LCLs. Only one of eight in vitro EBV-converted BL lines that shifted to a more LCL-like growth pattern expressed LMP at a high level. This line also reexpressed the HLA-A11 antigen that was undetectable in its EBV-negative progenitor. Our findings suggest that the typical BL cell phenotype is associated with low expression of both proteins. PMID- 3037522 TI - Human T-cell lymphotropic virus IIIB glycoprotein (gp120) bound to CD4 determinants on normal lymphocytes and expressed by infected cells serves as target for immune attack. AB - The lymphocyte differentiation antigen CD4 serves as a receptor for human retroviruses associated with acquired immunodeficiency syndrome (AIDS) through its interaction with the major envelope virion glycoprotein, gp120, which is also expressed on the surface of infected cells. In these experiments, purified gp120 was shown to bind to normal human T-lymphocyte populations. The gp120-CD4 complex served as a target antigen for antibody-dependent complement-mediated cytolysis by a goat serum raised against native gp120. However, patient sera that bound to gp120-adsorbed cells failed to direct their destruction in the presence of complement. In contrast, these sera were potent mediators of antibody-dependent cellular cytotoxicity. These studies demonstrate that gp120 situated on the cell surface can serve as an effective target for immune destruction by patient antibodies and effector lymphocytes. The possible contribution of this type of immunity to control of disease progression, on the one hand, and to lymphocyte destruction and immunopathology observed in AIDS, on the other, is discussed. PMID- 3037523 TI - Transposition of bacteriophage Mu in the Legionnaires disease bacterium. AB - Legionnaires disease is an acute respiratory disease that is often fatal for immunocompromised patients. The causative agent of this disease, Legionella pneumophila, is a Gram-negative bacterium that is present in a variety of aquatic environments. L. pneumophila is a facultative intracellular parasite; it grows within human phagocytic cells and eventually causes their destruction. In contrast to many other intracellular parasites, L. pneumophila is a Gram-negative bacterium that can be grown in standard microbiological culture medium. To determine the factors that enable this organism to enter, survive, and multiply within human mononuclear phagocytes, we chose bacteriophage Mu, a powerful genetic tool that transposes within the host cell genome, to generate insertion mutations and gene fusions in the Legionella genome. Certain derivatives of Mu are able to generate fusions between target genes and the lac operon from Escherichia coli. We have determined that although Mu is unable to attach to L. pneumophila or complete its life cycle within Legionella, it does transpose within the Legionella genome. Transposition was detected with a mini-Mu phage that carries the lac operon of E. coli. PMID- 3037524 TI - RepA and DnaA proteins are required for initiation of R1 plasmid replication in vitro and interact with the oriR sequence. AB - RepA, an initiation protein of R1 plasmid replication, was purified from an Escherichia coli strain overproducing the protein. The purified RepA protein specifically initiated replication in vitro of plasmid DNA bearing the replication origin of R1 plasmid (oriR). The replication, strictly dependent on added RepA protein, was independent of host RNA polymerase but required other host replication functions (DnaB and DnaC proteins, the single-stranded-DNA binding protein SSB, and DNA gyrase). The replication was also completely dependent on the host DnaA function. In filter binding assays in high salt (0.5 M KCl) conditions, RepA specifically binds to both supercoiled and linear plasmid DNA containing the oriR sequence, whereas it binds to nonspecific DNA in low salt. DNase I-protection studies on a linearized DNA fragment revealed that DnaA protein specifically binds to a 9-base-pair DnaA-recognition sequence ("DnaA box") within oriR only when RepA is bound to the sequence immediately downstream of the DnaA box. These results indicate that initiation of R1 plasmid replication is triggered by interaction of RepA and DnaA proteins with the oriR sequence. PMID- 3037525 TI - On the biologically active structures of cholecystokinin, little gastrin, and enkephalin in the gastrointestinal system. AB - The biologically active conformations of a series of four peptides [four cholecystokinin (CCK)-related peptides and enkephalin] in their interactions with gastrointestinal receptors have been deduced using conformational computational analysis. The two peptides that interact exclusively with peripheral-type CCK receptors are the heptapeptide COOH-terminal fragment from CCK (CCK-7) and the analogous sequence from cerulein (CER-7) in which threonine replaces the methionine proximal to the NH2 terminus. The two peptides that interact exclusively with the gastrin receptor in the stomach are the active COOH-terminal fragment of little gastrin and the COOH-terminal tetrapeptide sequence common to all of these peptides, CCK-4. We find that preferred conformations for the peripherally active peptides CCK-7 and CER-7 are principally beta-bends, whereas little gastrin and CCK-4 are fundamentally helical. In the class of lowest energy structures for both CCK-7 and CER-7, the aromatic rings of the tyrosine and phenylalanine lie close to one another whereas the tryptophan indole ring points in the opposite direction. This structure is superimposable on the structures of a set of rigid indolyl benzodiazepine derivatives that interact with complete specificity and high affinity with peripheral CCK receptors further suggesting that the computed beta-bends are the biologically active conformation. The biologically active conformation for CCK-4 and the little gastrin hexapeptide has also been deduced. By excluding conformations common to CCK-7 and CCK-4, which do not bond to each other's receptors, and then by selecting conformations in common to CCK-4 and the gastrin-related hexapeptide, which do bind to each other's receptors, we deduce that the biologically active conformation at the gastrin receptor is partly helical and one in which the indole of tryptophan and the aromatic ring of phenylalanine are close to one another while the methionine and aspartic acid side chains point in the opposite direction. These major differences in preferred structures between the common CCK-7/CER-7 peptides and the common CCK-4/little gastrin peptides explain the mutually exclusive activities of these two sets of peptides. We have observed that [Met]enkephalin strongly antagonizes the action of the naturally occurring peripherally active CCK-8 (CCK-7 with an NH2-terminal aspartic acid residue added). The computed lowest energy structures for this opiate peptide closely resemble key features of the computed CCK-7/CER-7 structure, further supporting the proposed structure. PMID- 3037526 TI - Molecular cloning and sequence analysis of the catalytic subunit of bovine type 2A protein phosphatase. AB - We have isolated a cDNA clone corresponding to the Mr 38,000 catalytic subunit of bovine type 2A protein phosphatase. The cDNA was isolated from a bovine adrenal gland cDNA library through the use of oligonucleotide probes whose sequences were based on partial amino acid sequence obtained from cyanogen bromide fragments of the purified cardiac enzyme. The entire 1724-base-pair cDNA has been sequenced and found to contain an open reading frame coding for a protein of 325 amino acids having a calculated molecular weight of 37,320. The deduced amino acid sequence contains the experimentally determined sequences of five different cyanogen bromide peptides. Transfection of COS-m6 cells with the cloned cDNA resulted in transient expression of a protein that could be detected by immunoblot analysis with a monoclonal antibody directed against the purified cardiac protein phosphatase. The expressed protein had the same apparent molecular weight as the purified enzyme when analyzed by NaDodSO4/polyacrylamide gel electrophoresis, suggesting that this clone contains the entire coding region of the phosphatase mRNA. The cloned cDNA hybridizes to a mRNA of 2.0 kilobases in bovine heart and adrenal gland. Under conditions of reduced stringency, the cDNA also hybridizes to a mRNA species of 1.2 kilobases in cardiac tissue. PMID- 3037527 TI - Multiple sequence elements are required for regulation of human T-cell leukemia virus gene expression. AB - The U3 region of the long terminal repeat (LTR) of human T-cell leukemia virus type I (HTLV-I) contains sequences that respond to the trans-activating transcription (tat) factor encoded by the pX region of the provirus. Results presented here show that there are multiple tat-responsive sequences within the LTR and that a single 21-nucleotide sequence, which is repeated three times within the U3 region, is sufficient to determine the response to the trans activator. This sequence is capable of conferring a tat-responsive phenotype upon the HTLV-I and simian virus 40 promoters, independent of orientation. Sequences required for efficient HTLV-I LTR-directed gene expression are also located 3' to the site of RNA initiation, within the R and U5 regions of the LTR. PMID- 3037528 TI - An extrachromosomal form of the Mu transposons of maize. AB - Maize lines known as Robertson's Mutator (Mu) lines generate unstable recessive mutations at high frequencies. These lines carry actively transposing copies of the transposons (Tn) Mu1 and Mu1.7. TnMu1 and TnMu1.7 are approximately 1400 and 1700 base pairs long, respectively, and they have 210-base-pair terminal inverted repeats. We report here extrachromosomal forms of TnMu1 and TnMu1.7. The extrachromosomal Mu1 and Mu1.7 molecules are resistant to alkaline denaturation and to proteinase treatment and have circular restriction maps; therefore, they are probably covalently closed circular DNA. Further, we show that their occurrence is correlated with Mu activity, so they are probably generated during Mu transposition as transposition intermediates or as products of Mu excision. When the total extrachromosomal supercoiled DNA from immature male flowers of a Mu line was examined by electron microscopy, the Mu transposons appeared to constitute a significant fraction of the extrachromosomal DNA circles in Mu lines. PMID- 3037529 TI - Sequence-specific interactions of nuclear factors with conserved sequences of human class II major histocompatibility complex genes. AB - All class II major histocompatibility complex genes contain two highly conserved sequences, termed X and Y, within the promoter regions(s), which may have a role in regulation of expression. To study trans-acting factors that interact with these sequences, sequence-specific DNA binding activity has been examined by the gel electrophoresis retardation assay using the HLA-DQ2 beta gene 5' flanking DNA and nuclear extracts derived from various cell types. Several specific protein binding activities were found using a 45-base-pair (bp) HinfI/Sau96I (-142 to -98 bp) and a 38-bp Sau96I/Sau96I (-97 to -60 bp) fragment, which include conserved sequence X (-113 to -100 bp) and conserved sequence Y (-80 to -71 bp), respectively. Competition experiments, methylation interference analysis, and DNase I foot-printing demonstrated that distinct proteins in a nuclear extract of Raji cells (a human B lymphoma line) bind to sequence X, to sequence Y, and to DNA 5' of the X sequence (termed sequence W). The factor binding site in the W sequence is also found to be conserved among beta-chain genes and is suggested to be a gamma-interferon control region. PMID- 3037530 TI - Modification of DNA ends can decrease end joining relative to homologous recombination in mammalian cells. AB - In animal cells, exogenous DNA recombines into random chromosomal sites much more frequently than it recombines into homologous sites. Free DNA ends are "recombinogenic" in both processes. To test the effects of specific ends on analogous extrachromosomal processes, we constructed a linear genome of simian virus 40 with terminal repeated sequences. After transfection into monkey cells, the model substrate can circularize by end joining (analogous to random integration) or by homologous recombination between its terminal repeats (analogous to targeted recombination). Since the two types of recombination are in competition with one another, the ratio of homologous-recombination to end join products is a sensitive indicator of the differential effects of specific ends. Substrates with blunt ends, complementary sticky ends, or mismatched ends generated the same ratio of homologous-recombination to end-join products. However, addition of dideoxynucleotides to the 3' hydroxyls of the substrate decreased the frequency of end joining by a factor of 5-6 relative to homologous recombination. Thus, the frequency of end joining can be decreased relative to that of homologous recombination by modification of the ends of the input DNA. These results suggest an approach to altering the ratio of random to targeted integration in mammalian cells. PMID- 3037531 TI - Molecular cloning of the akt oncogene and its human homologues AKT1 and AKT2: amplification of AKT1 in a primary human gastric adenocarcinoma. AB - A previous report described the isolation of a directly transforming retrovirus, AKT8, from a spontaneous thymoma of an AKR mouse. The AKT8 provirus has now been molecularly cloned from a transformed, nonproducer cell line. The virus genome contains both viral and nonviral, cell-related sequences; the nonviral sequence has been designated v-akt, the presumed viral oncogene of the AKT8 virus. This gene lacks homology to the 16 other oncogenes tested. The cloned provirus has undergone a partial deletion, during cell passage in vitro, that prevents direct demonstration of the transforming ability of this molecular clone. Two human homologues of the v-akt oncogene, AKT1 and AKT2, were cloned. A survey of 225 human tumors for changes involving AKT1 led to the discovery of a 20-fold amplification of this gene in one of the five gastric adenocarcinomas tested. The results demonstrate that AKT8 has the characteristic structure of a directly transforming retrovirus and that it contains a gene derived from highly conserved cellular sequences that may be involved in the pathogenesis of some human malignancies. PMID- 3037532 TI - Phencyclidine is a negative allosteric modulator of signal transduction at two subclasses of excitatory amino acid receptors. AB - Phencyclidine (PCP) and some of its pharmacological congeners inhibit the signal transduction at specific excitatory amino acid receptors of cerebellar granule cells in primary cultures. These drugs do not bind to the transmitter recognition sites, and affinity of this specific binding site is increased by the presence of the transmitter bound to its recognition sites. PCP inhibits phosphatidylinositol phosphate hydrolysis mediated by Mg2+-sensitive glutamate receptors (GP1) but not that mediated by Mg2+-insensitive glutamate receptors (GP2). In addition, PCP inhibits Ca2+ influx and cGMP formation mediated by the activation of Mg2+ sensitive glutamate receptors (GC1) but not that mediated by Mg2+-insensitive glutamate receptors (GC2). In this cell culture the activation of phosphatidylinositol phosphate hydrolysis by muscarinic receptor agonists is not affected by PCP. Since PCP inhibits noncompetitively GP1 and GC1 signal transduction it may act as a negative allosteric modulator of signal transduction at both receptors. The pharmacological profile of PCP and its congeners delimits a class of drugs modulating allosterically the action of the primary transmitter at GP1 and GC1 receptors. These drugs need the presence of the transmitter to act and they cannot be termed inverse agonists because they are devoid of activity in the absence of the transmitter; moreover, they do not bind to the transmitter recognition site nor do they prevent the transmitter binding to its recognition sites. PMID- 3037533 TI - Localization of beta-adrenergic receptors on differentiated cells of the central nervous system in culture. AB - Hypothalamic neurons and cortical protoplasmic (type 1) astrocytes in dissociated cultures of rat brain express binding sites for antibodies specific for epitopes related to the beta-adrenergic receptor. Undifferentiated glial progenitor cells of the rat optic nerve do not detectably bind these antibodies, but both of the progeny [oligodendroglia and process-bearing (type 2) astrocytes] express immunologically identified beta-adrenergic receptors. Levels of receptor expression are variable: not all cells of each type express receptors nor is expression uniform on a given cell. The data suggest that cells of the central nervous system express beta-adrenergic receptors but that levels of expression may be determined by the differentiated state of the cells. PMID- 3037534 TI - Nucleotide sequence and expression of the mercurial-resistance operon from Staphylococcus aureus plasmid pI258. AB - The mercurial-resistance determinant from Staphylococcus aureus plasmid pI258 is located on a 6.4-kilobase-pair Bgl II fragment. The determinant was cloned into both Bacillus subtilis and Escherichia coli. Mercury resistance was found only in B. subtilis. The 6404-base-pair DNA sequence of the Bgl II fragment was determined. The mer DNA sequence includes seven open reading frames, two of which have been identified by homology with the merA (mercuric reductase) and merB (organomercurial lyase) genes from the mercurial-resistance determinants of Gram negative bacteria. Whereas 40% of the amino acid residues overall were identical between the pI258 merA polypeptide product and mercuric reductases from Gram negative bacteria, the percentage identity in the active-site positions and those thought to be involved in NADPH and FAD contacts was above 90%. The 216 amino acid organomercurial lyase sequence was 39% identical with that from a Serratia plasmid, with higher conservation in the middle of the sequences and lower homologies at the amino and carboxyl termini. The remaining five open reading frames in the pI258 mer sequence have no significant homologies with the genes from previously sequenced Gram-negative mer operons. PMID- 3037535 TI - Down-regulation of rat kidney calcitonin receptors by salmon calcitonin infusion evidenced by autoradiography. AB - In treating age-related osteoporosis and Paget disease of bone, it is of major importance to avoid an escape phenomenon that would reduce effectiveness of the treatment. The factors involved in the loss of therapeutic efficacy with administration of large pharmacological doses of the hormone require special consideration. Down-regulation of the hormone receptors could account for the escape phenomenon. Specific binding sites for salmon calcitonin (sCT) were characterized and localized by autoradiography on rat kidney sections incubated with 125I-labeled sCT. Autoradiograms demonstrated a heterogenous distribution of 125I-labeled sCT binding sites in the kidney, with high densities in both the superficial layer of the cortex and the outer medulla. Infusion of different doses of unlabeled sCT by means of Alzet minipumps for 7 days produced rapid changes in plasma calcium, phosphate, and magnesium levels, which were no longer observed after 2 or 6 days of treatment. Besides, infusion of high doses of sCT induced down-regulation of renal sCT binding sites located mainly in the medulla, where calcitonin (CT) has been shown to exert its physiological effects on water and ion reabsorption. These data suggest that the resistance to high doses of sCT often observed during long-term treatment of patients may be the consequence of not only bone-cell desensitization but also down-regulation of CT-sensitive kidney receptor sites. PMID- 3037536 TI - Isolation of cDNA clones coding for human tissue factor: primary structure of the protein and cDNA. AB - Tissue factor is a membrane-bound procoagulant protein that activates the extrinsic pathway of blood coagulation in the presence of factor VII and calcium. lambda Phage containing the tissue factor gene were isolated from a human placental cDNA library. The amino acid sequence deduced from the nucleotide sequence of the cDNAs indicates that tissue factor is synthesized as a higher molecular weight precursor with a leader sequence of 32 amino acids, while the mature protein is a single polypeptide chain composed of 263 residues. The derived primary structure of tissue factor has been confirmed by comparison to protein and peptide sequence data. The sequence of the mature protein suggests that there are three distinct domains: extracellular, residues 1-219; hydrophobic, residues 220-242; and cytoplasmic, residues 243-263. Three potential N-linked carbohydrate attachment sites occur in the extracellular domain. The amino acid sequence of tissue factor shows no significant homology with the vitamin K-dependent serine proteases, coagulation cofactors, or any other protein in the National Biomedical Research Foundation sequence data bank (Washington, DC). PMID- 3037537 TI - Nucleotide sequence of the gene coding for human factor VII, a vitamin K dependent protein participating in blood coagulation. AB - Activated factor VII (factor VIIa) is a vitamin K-dependent plasma serine protease that participates in a cascade of reactions leading to the coagulation of blood. Two overlapping genomic clones containing sequences encoding human factor VII were isolated and characterized. The complete sequence of the gene was determined and found to span about 12.8 kilobases. The mRNA for factor VII as demonstrated by cDNA cloning is polyadenylylated at multiple sites but contains only one AAUAAA poly(A) signal sequence. The mRNA can undergo alternative splicing, forming one transcript containing eight segments as exons and another with an additional exon that encodes a larger prepro leader sequence. The latter transcript has no known counterpart in the other vitamin K-dependent proteins. The positions of the introns with respect to the amino acid sequence encoded by the eight essential exons of factor VII are the same as those present in factor IX, factor X, protein C, and the first three exons of prothrombin. These exons code for domains generally conserved among members of this gene family. The comparable introns in these genes, however, are dissimilar with respect to size and sequence, with the exception of intron C in factor VII and protein C. The gene for factor VII also contains five regions made up of tandem repeats of oligonucleotide monomer elements. More than a quarter of the intron sequences and more than a third of the 3' untranslated portion of the mRNA transcript consist of these minisatellite tandem repeats. PMID- 3037538 TI - The molecular cloning of a type II regulatory subunit of the cAMP-dependent protein kinase from rat skeletal muscle and mouse brain. AB - A cDNA clone for a type II regulatory (R) subunit of the cAMP-dependent protein kinase (ATP:protein phosphotransferase, EC 2.7.1.37) was isolated from a rat skeletal muscle library using a specific 47-base oligonucleotide probe. The rat cDNA was 1.2 kilobases (kb) in length and contained an open reading frame of 1.113 kb representing 92% of the coding region of the molecule. Nick-translated rat cDNA was then used to isolate a mouse RII cDNA clone from a brain library that contained an open reading frame of 1.143 kb. Because both cDNAs lacked complete coding sequences, the remainder of the RII coding region was obtained from a 15-kb mouse genomic clone. The mouse RII coding region contains 1.2 kb corresponding to a 400-amino acid protein of 51.141 kDa. The mouse cDNA hybridizes to two mRNA species, a 2.4-kb form that was only observed in testis and a 6.0-kb form found in a wide range of tissues, including testis. PMID- 3037539 TI - Functional analysis of a retroviral host-range mutant: altered long terminal repeat sequences allow expression in embryonal carcinoma cells. AB - A retroviral host-range neomycin-resistant myeloproliferative sarcoma virus mutant, which is expressed in the embryonal carcinoma cell lines F9 and PCC4aza1R, was molecularly cloned and analyzed. This mutant virus, PCMV, differs from myeloproliferative sarcoma virus by two major deletions, one of which spans exactly a 75-base-pair repeat of the long terminal repeat. Functional analysis of recombinant viruses shows that the host-range expansion of PCMV is a property of nucleotide changes within the U3 region of the long terminal repeat. Furthermore, expression assays of chimeric long terminal repeats show that the enhancer region of PCMV joined to the promoter region of Moloney murine leukemia virus is sufficient to direct the synthesis of chloramphenicol acetyltransferase in F9 and PCC4 cells. PMID- 3037540 TI - Regulation of cell shape in the Cloudman melanoma cell line. AB - We show that Cloudman melanoma cells undergo rapid arborization in response to [Nle4,D-Phe7]alpha-melanocyte-stimulating hormone, a potent analogue of alpha melanocyte stimulating hormone (alpha-MSH). The arbors were established by extension of processes and resembled dendrites. We used this system to study the regulation of cell shape. alpha-MSH is known to induce increases in cAMP levels, and agents such as forskolin and isobutylmethylxanthine that led to increased cAMP also caused arborization. However, equally dramatic arbors were formed after incubation with the protein kinase C inhibitor H-7 [1-(5-isoquinolinesulfonyl) alpha-methyl-piperazine]. Phorbol diesters that activate protein kinase C led to cell rounding and antagonized alpha-MSH. The actions of protein kinase C cannot be rationalized in terms of indirect effects on cAMP: neither H-7 nor phorbol diesters alone altered cAMP levels, nor did they affect the increase in cAMP induced by MSH. We show also that MSH produced longer-term effects that cannot be mimicked by cAMP. Specifically, even in the continued presence of alpha-MSH, arborization was followed by morphological reversal to the unstimulated flattened configuration within 2 hr. (This did not occur with other agents that increase cAMP or with H-7.) Most importantly, whereas MSH-induced arborization occurred in the presence of cycloheximide, actinomycin D, or in enucleated cells, the reversal of arborization did not. Thus, MSH induced a program of rapid shape change that was dependent on new protein synthesis and gene transcription. PMID- 3037541 TI - Spermatid-specific expression of protamine 1 in transgenic mice. AB - Protamines are abundant basic proteins involved in the condensation of sperm chromatin. In the mouse, protamine genes are transcribed postmeiotically in round spermatids. We have cloned and sequenced the mouse protamine 1 gene. Ten lines of transgenic mice harboring marked protamine 1 sequences were generated by microinjection of fertilized eggs. Transcription of the transgene is restricted to round spermatids and in several cases exceeds that of the endogenous gene. The cis-acting sequences required for tissue-specific protamine expression reside on a 2.4-kilobase restriction fragment. Prospects for using transgenic mice to address fundamental questions of male germ-cell development are discussed. PMID- 3037542 TI - Isolation and characterization of a nematode transposable element from Panagrellus redivivus. AB - We have isolated a transposable element, designated PAT-1, from the free-living nematode Panagrellus redivivus. P. redivivus strain C15 was found to have a high spontaneous mutation frequency compared to the standard Caenorhabditis elegans laboratory strain N2. To characterize the genetic lesions occurring in spontaneous C15 mutants, we molecularly cloned the homolog of the C. elegans unc 22 gene from wild-type P. redivivus and two strains carrying spontaneous mutations in this gene. One of these mutations resulted from the insertion of a 4.8-kilobase segment of repetitive DNA. This repetitive element (PAT-1) varies in copy number (10-50 copies) and location in different P. redivivus strains and is absent from C. elegans. The element could be useful as a transformation vector for C. elegans. Our approach is a general one that could be used to isolate additional nematode transposons from other species. PMID- 3037543 TI - Characterization of a panel of somatic cell hybrids for regional mapping of the mouse X chromosome. AB - A panel of five hybrid cell lines containing mouse X chromosomes with various deletions has been obtained by fusing splenocytes from male mice carrying one of a series of reciprocal X-autosome translocations with the azaguanine-resistant Chinese hamster cell line CH3g. These hybrids have been extensively characterized by using the allozymes hypoxanthine/guanine phosphoribosyltransferase (encoded by the Hprt locus) and alpha-galactosidase (Ags) and a series of 11 X-chromosome specific DNA probes whose localization had been previously established by linkage studies. Such studies have established the genetic breakpoints of the T(X;12)13Rl and T(X;2)14Rl X-autosome translocations on the X chromosome and provided additional information as to the X-chromosome genetic breakpoints of the T(X;16)16H, T(X;4)7Rl, and T(X;7)6Rl translocations. The data establish clearly that both the T(X;4)7Rl and T(X;12)13Rl X-chromosome breakpoints are proximal to Hprt, the breakpoint of the former being more centromeric, lying as it does in the 9-centimorgan interval between the ornithine transcarbamoylase (Otc) and DXPas7 (M2C) loci. Similarly, it is now clear that the T(X;16)16H X-autosome translocation breakpoint lies distal to the DXPas8 (St14-1) locus, narrowing the X-chromosome breakpoint down to a region flanked proximally by this marker and representing, as expected from previous data, the distal quarter of the Hprt-Ta subchromosomal span. These five hybrid cell lines provide, with the previously characterized EBS4 hybrid cell line, a nested series of seven mapping intervals distributed along the length of the mouse X chromosome. Their characterization not only allows further correlation of the genetic and cytological X-chromosome maps but also should permit the rapid identification of DNA probes specific for particular regions of the mouse X chromosome. PMID- 3037544 TI - Differential effects of base-pair mismatch on intrachromosomal versus extrachromosomal recombination in mouse cells. AB - To initially determine the effect that base-pair mismatch has on homologous recombination in mammalian cells, we have studied genetic recombination between thymidine kinase (tk) gene sequences from herpes simplex virus 1 and 2. These tk genes are approximately 81% homologous at the nucleotide level. We observed that, in mouse LTK- cells, intrachromosomal recombination between type 1 and type 2 tk sequences is reduced by a factor of at least 1000 relative to the rate of intrachromosomal recombination between homologous type 1 tk sequences. In sharp contrast, the rate of intermolecular or intramolecular extrachromosomal recombination between the heterologous tk sequences introduced by calcium phosphate or microinjection was reduced only by a factor of 3 to 15 compared with extrachromosomal homologous tk crosses. Our results suggest differences between the mechanisms of extrachromosomal and intrachromosomal recombination in mammalian cells. PMID- 3037545 TI - HRAS1-selected chromosome transfer generates markers that colocalize aniridia- and genitourinary dysplasia-associated translocation breakpoints and the Wilms tumor gene within band 11p13. AB - We show that chromosome-mediated gene transfer can provide an enriched source of DNA markers for predetermined, subchromosomal regions of the human genome. Forty four human DNA recombinants isolated from a HRAS1-selected chromosome-mediated gene transformant map exclusively to chromosome 11, with several sublocalizing to the Wilms tumor region at 11p13. We present a detailed molecular map of the deletion chromosomes 11 from five WAGR (Wilms tumor/aniridia/genitourinary abnormalities/mental retardation) syndrome patients, three of which are at the limits of cytogenetic resolution but shown here to be molecularly distinguishable and overlapping. We can define ten distinct regions of the short arm of chromosome 11, five of which subdivide band 11p13. We also map two independent 11p13 translocation breakpoints to within the smallest region of overlap defined by the WAGR deletions. The first comes from a patient with familial aniridia, and the second from a patient with Potter facies and genitourinary dysplasia. The close similarities in map location and affected cell lineage for Wilms tumor and genitourinary dysplasia suggest that they may be alternative manifestations of mutation at the same locus. PMID- 3037546 TI - Alternative use of 5' exons in the specification of Ly-5 isoforms distinguishing hematopoietic cell lineages. AB - Previous inferences that Ly-5 glycoprotein isoforms of murine hematopoietic cells are generated by alternative splicing of primary transcripts of a single Ly-5 gene are supported by the present study. A cDNA library was prepared from B cells by extension from primer representing a known T-cell cDNA sequence. Three different Ly-5 clones from this library included sequences missing in T-cell cDNA clones. From the constitution of cDNA clones and of the Ly-5 gene, and from S1 nuclease mapping, it is concluded that at least two exons, provisionally numbered Ex-6(B) and Ex-7(B), in the 5'-proximal region are mainly represented in mRNA of the B-cell lines examined but not of the T-cell lines examined. Also, exons 1 and 2 appear to be used alternatively in different species of B-cell mRNA and probably also in different species of T-cell mRNA. PMID- 3037547 TI - Generation of specific cytotoxic T cells with a fragment of the Epstein-Barr virus-encoded p63/latent membrane protein. AB - Human B lymphocytes, transformed by the herpesvirus Epstein-Barr virus, are known to express a characteristic antigen(s) recognized by the cellular immune response. This structure has been termed lymphocyte-determined membrane antigen. Because of the significance of this structure in controlling Epstein-Barr virus infection in vivo, the molecular nature of lymphocyte-determined membrane antigen has been long sought. In this paper, we show that a sequence of 10 amino acids (residues 43-53) from the Epstein-Barr virus-encoded membrane protein p63/latent membrane protein can induce Epstein-Barr virus-specific cytotoxic T cells and, therefore, bears at least one of the lymphocyte-determined membrane antigenic determinants. PMID- 3037548 TI - Activation of interleukin 2 and interleukin 2 receptor (Tac) promoter expression by the trans-activator (tat) gene product of human T-cell leukemia virus, type I. AB - Cotransfection of cDNA encoding the trans-activator gene product of human T-cell leukemia virus, type I (HTLV-I) (tat-I), which acts in trans to augment viral gene expression, has revealed strong regulatory effects of this viral protein on the inducible cellular promoters governing human interleukin 2 (IL-2) and IL-2 receptor (Tac) gene expression. The tat-I protein stimulates a 3- to 6-fold increase in IL-2 receptor (Tac) promoter activity in transfected Jurkat T cells, but not in the natural killer-like YT cell line, as measured by changes in the expression of the chloramphenicol acetyltransferase (CAT; EC 2.3.1.28) reporter gene linked to this promoter. In contrast, tat-I alone has little or no effect on IL-2 promoter activity in Jurkat T cells but markedly synergizes with other mitogenic stimuli (phytohemagglutinin, phorbol 12-myristate 13-acetate, or the OKT3 monoclonal antibody), which alone are ineffective. The tat-I protein also partially circumvents the pronounced inhibitory effects of cyclosporin A on the IL-2 promoter. Other cellular and viral promoters are unaffected by the tat-I gene product, either alone or in combination with other mitogens. The specific effects of the tat-I gene product on the IL-2 and IL-2 receptor (Tac) promoters suggest the possibility of an autocrine or paracrine mechanism of T-cell growth as an early event in HTLV-I-mediated leukemogenesis. PMID- 3037549 TI - Cannabinoids induce incomplete maturation of cultured human leukemia cells. AB - Monocyte maturation markers were induced in cultured human myeloblastic ML-2 leukemia cells after treatment for 1-6 days with 0.03-30 microM delta 9 tetrahydrocannabinol (THC), the major psychoactive component of marijuana. After a 2-day or longer treatment, 2- to 5-fold increases were found in the percentages of cells exhibiting reactivity with either the murine OKM1 monoclonal antibody or the Leu-M5 monoclonal antibody, staining positively for nonspecific esterase activity, and displaying a promonocyte morphology. The increases in these differentiation markers after treatment with 0.03-1 microM THC were dose dependent. At this dose range, THC did not cause an inhibition of cell growth. The THC-induced cell maturation was also characterized by specific changes in the patterns of newly synthesized proteins. Pronounced among these changes was an increase in the synthesis of at least 10 proteins that are found abundantly in monocytes. The THC-induced differentiation did not, however, result in cells with a highly developed mature monocyte phenotype; the THC-treated cells failed to exhibit other monocyte markers such as attachment to the surface of tissue culture dishes or morphological maturation beyond the promonocyte stage. However, treatment of these "incompletely" matured cells with either phorbol 12-myristate 13-acetate or 1 alpha,25-dihydroxycholecalciferol, which are inducers of differentiation in myeloid leukemia cells (including ML-2 cells), produced cells with a mature monocyte morphology. Two other cannabinoids, cannabidiol and cannabinol, which were more cytotoxic than THC at comparable doses, also caused an increase in the expression of maturation markers, but at doses higher than those required for THC. The ML-2 cell system described here may be a useful tool for deciphering critical biochemical events that lead to the cannabinoid-induced "incomplete" cell differentiation of ML-2 cells and other related cell types. Findings obtained from this system may have important implications for studies of cannabinoid effects on normal human bone-marrow progenitor cells. PMID- 3037550 TI - Molecular genetic approach to human meningioma: loss of genes on chromosome 22. AB - A molecular genetic approach employing polymorphic DNA markers has been used to investigate the role of chromosomal aberrations in meningioma, one of the most common tumors of the human nervous system. Comparison of the alleles detected by DNA markers in tumor DNA versus DNA from normal tissue revealed chromosomal alterations present in primary surgical specimens. In agreement with cytogenetic studies of cultured meningiomas, the most frequent alteration detected was loss of heterozygosity on chromosome 22. Forty of 51 patients were constitutionally heterozygous for at least one chromosome 22 DNA marker. Seventeen of the 40 constitutionally heterozygotic patients (43%) displayed hemizygosity for the corresponding marker in their meningioma tumor tissues. Loss of heterozygosity was also detected at a significantly lower frequency for markers on several other autosomes. In view of the striking association between acoustic neuroma and meningioma in bilateral acoustic neurofibromatosis and the discovery that acoustic neuromas display specific loss of genes on chromosome 22, we propose that a common mechanism involving chromosome 22 is operative in the development of both tumor types. Fine-structure mapping to reveal partial deletions in meningiomas may provide the means to clone and characterize a gene (or genes) of importance for tumorigenesis in this and possibly other clinically associated tumors of the human nervous system. PMID- 3037551 TI - Interaction between the human T-cell lymphotropic virus type IIIB envelope glycoprotein gp120 and the surface antigen CD4: role of carbohydrate in binding and cell fusion. AB - Interactions between retroviruses associated with acquired immunodeficiency syndrome and their receptors on lymphocytes represent the initial steps in the process of infection and are also involved in multinucleated giant cell formation, which is one form of virus-mediated cytopathology. The exterior envelope glycoprotein of the retrovirus has been identified as gp120, and we demonstrate here that purified gp120 binds directly to cells expressing the CD4 (T4) surface antigen at a site spatially related to that recognized by the OKT4A monoclonal antibody. The gp120 was also able to temporarily interfere with viral infection and to block the process of multinucleated giant cell formation. However, if the carbohydrate chains were removed from gp120 by enzymatic treatment, CD4 binding and blockade of cell fusion was reduced by about a factor of 50. The significance of these results in relation to preventive and interventive approaches for acquired immunodeficiency syndrome is discussed. PMID- 3037552 TI - Anti-glycoprotein D antibodies that permit adsorption but block infection by herpes simplex virus 1 prevent virion-cell fusion at the cell surface. AB - Certain monoclonal antibodies specific for glycoprotein D of herpes simplex virus have potent neutralizing activity but fail to block attachment of virus to cells. Here we have investigated the fate of neutralized and infectious virus after attachment to primate cells. Infectious virions fused with the cell surface such that naked nucleocapsids were detectable in the cytoplasm near or just under the plasma membrane. Neutralized virions did not fuse with the cell. They remained attached to the cell surface and could be rendered infectious by treatment with polyethylene glycol. We conclude that some anti-glycoprotein D neutralizing antibodies can inhibit the penetration of herpes simplex virus by blocking fusion of the virion envelope with the plasma membrane. These results identify a pathway of entry that initiates successful herpes simplex virus infection and a step in this pathway that is highly sensitive to neutralizing antibodies. A role for glycoprotein D in virion-cell fusion is indicated. PMID- 3037553 TI - Simultaneous measurement of hormone release and secretagogue binding by individual pituitary cells. AB - The quantitative relationship between receptor binding and hormone secretion at the single-cell level was investigated in the present study by combining a reverse hemolytic plaque assay for measurement of luteinizing hormone (LH) secretion from individual pituitary cells with an autoradiographic assay of 125I labeled gonadotropin-releasing hormone (GnRH) agonist binding to the same cells. In the plaque assay, LH secretion induces complement-mediated lysis of the LH antibody-coated erythrocytes around the gonadotropes, resulting in areas of lysis (plaques). LH release from individual gonadotropes was quantified by comparing radioimmunoassayable LH release to hemolytic area in similarly treated cohort groups of cells; plaque area was linearly related to the amount of LH secreted. Receptor autoradiography was performed using 125I-labeled GnRH-A (a superagonist analog of GnRH) both as the ligand and as the stimulant for LH release in the plaque assay; the developed silver grains appearing over cells in the center of plaques were measured microscopically. The grains appeared to represent specific and high-affinity receptors for GnRH because no pituitary cells other than gonadotropes bound the labeled ligand and grain development was progressively inhibited by coincubation with increasing doses of unlabeled GnRH-A. Despite high correlations between mean grain number and mean plaque area in dose-response curves, the correlation coefficients for these parameters were low (range 0.02 0.38) in the individual cells comprising these groups. We conclude that GnRH receptor number for any individual gonadotrope is a weak determinant of the amount of LH it can secrete; nevertheless, full occupancy of all its GnRH receptors is required for any gonadotrope to reach its full LH-secretory capacity. Apparently the levels of other factors comprising the steps along the secretory pathway determine the secretory capacity of an individual cell. PMID- 3037554 TI - Additive beneficial effects of two inhibitors of vasoconstrictor mediators in acute myocardial ischemia. AB - A specific inhibitor of thromboxane A2 (TxA2) synthesis, CGS-12970, a new angiotensin-converting enzyme (ACE) inhibitor, CGS-16617, and a combination of both agents were evaluated for their ability to reduce the extension of myocardial infarct size in rats. Myocardial creatine kinase (CK) loss from the left ventricular free wall (LVFW) 48 hr after left coronary artery ligation was used as an index of ischemic damage. Treatment with either CGS-12970 (4 mg/kg) or CGS-16617 (1 microgram/kg) alone did not attenuate the loss of CK from LVFW significantly, compared with animals receiving only the vehicles for these drugs. However, the combined use of both agents significantly reduced CK depletion from LFVW (P less than 0.01). These findings support the interrelated role of TxA2 and angiotensin II as mediators of myocardial ischemia and suggest that combined inhibition of their formation may be useful in the treatment of acute myocardial infarction. PMID- 3037555 TI - Sex differences in n-3 and n-6 fatty acid metabolism in EFA-depleted rats. AB - We studied the effect of sex on the distribution of long-chain n-3 and n-6 fatty acids in essential fatty acid-deficient rats fed gamma-linolenate (GLA) concentrate and/or eicosapentaenoate and docosahexaenoate-rich fish oil (FO). Male and female weanling rats were rendered essential fatty acid deficient by maintaining them on a fat-free semisynthetic diet for 8 weeks. Thereafter, animals of each sex were separated into three groups (n = 6) and given, for 2 consecutive days by gastric intubation, 4 g/kg body wt per day of GLA concentrate (containing 84% 18:2n-6), n-3 fatty acid-rich FO (containing 18% 20:5n-3 and 52% 22:6n-3), or an equal mixture of the two oil preparations (GLA + FO). The fatty acid distributions in plasma and liver lipids were then examined. GLA treatment increased the levels of C-20 and C-22 n-6 fatty acids in all lipid fractions indicating that GLA was rapidly metabolized. However, the increases in 20:3n-6 were less in females than those in males, while those in 20:4n-6 were greater, suggesting that the conversion of 20:3n-6 to 20:4n-6 was more active in female than in male rats. FO treatment increased the levels of 20:5n-3 and 22:6n-3 and reduced those of 20:4n-6. The increase in n-3 fatty acids was greater in females than that in males and the reduction in 20:4n-6 was smaller. Consequently, the sum of total long-chain EFAs incorporated was greater in females than that in males. The administration of n-3 fatty acids also reduced the ratio of 20:4n-6 to 20:3n-6 in GLA + FO-treated rats indicating that n-3 fatty acids inhibited the activity of delta-5-desaturase. However, this effect was not affected by the sex difference. PMID- 3037556 TI - Biochemical characterization of mutants in cyclic nucleotide metabolism showing rhythmic conidiation in Neurospora. AB - Mutants, cpd-1, cpd-2, and bd, which have reduced concentrations of orthophosphate-repressible cyclic phosphodiesterase (cPDase), show reduced concentrations of cyclic 3',5'-AMP and exhibit rhythmic oscillations of cyclic 3',5'-AMP with about 24-hr period length. These mutants have either reduced concentrations of adenylate cyclase or enhanced concentrations of Mg-stimulated cyclic phosphodiesterase. These mutations changed the ability to activate heat activated cyclic phosphodiesterase. White light reduced the concentration of cyclic 3',5'-GMP in mycelia. PMID- 3037557 TI - Cell typing and connections of the rat suprachiasmatic nucleus. AB - Microiontophoretic delivery of horseradish peroxidase and Lucifer Yellow liposomes into several nuclei (arcuate nucleus, median eminence, ventromedial hypothalamic nucleus, and retrochiasmatic area) of the medial basal hypothalamus revealed an intrinsic pattern of connections to the suprachiasmatic nucleus that was correlated to immunocytochemistry (LH-RH, leuenkephalin, and ACTH) of the fibers and cell types. Bilateral projections of the suprachiasmatic nucleus into the arcuate nucleus were found. The number of connections between the supraoptic nucleus and nuclei of the mediobasal hypothalamus was found to be greater than hitherto reported. A direct projection from the suprachiasmatic nucleus into the median eminence could not be demonstrated with the retrograde horseradish peroxidase and the Lucifer Yellow liposome technique. Intermediobasal hypothalamic connections are not only organized over the third ventricle but also ventrally through the median eminance. ACTH-like fibers penetrate the suprachiasmatic nucleus. These connections originate in the arcuate nucleus of the mediobasal hypothlamus. Leu-enkephalin was demonstrated within the center of the suprachiasmatic nucleus; LH-RH positive cells, however, surround the suprachiasmatic nucleus. PMID- 3037558 TI - Rhythmic pineal-hypophyseal-adrenal intermodulations ex vivo. AB - To assess circadian and circaseptan rhythmic intermodulation, female mice, kept on regimens of 12 hr light (L) alternating with 12 hr of darkness (D) (LD 12:12) were sampled for 1-13 days at six circadian stages. Pineals were removed and homogenized with 0.9% NaCl. Adenohypophyses were bisected and incubated in Krebs Ringer buffer (KRb) and/or with 10(-7) M melatonin (Mt). Adrenals were bisected and incubated with KRb, with 10(-7) M Mt, with 0.05 IU of ACTH 1-17 (Sy; Hoechst, Italy), or with Sy + 0.025 ml of the aqueous pineal homogenate (APH). ACTH and prolactin (PRL) in adenohypophysis incubation fluid were determined by radioimmunoassay (RIA), and corticosterone in adrenal incubation fluid was determined by RIA or by a fluorometric method. By this chronobiologic approach, time series were collected systematically in the search for predictable variability. Once these series reveal an algorithmically formulatable pattern, with a waveform validated by inferential statistical means, they are graded in terms of complexity as alpha, beta, gamma, and delta rhythms. Spontaneously (alpha-) rhythmic along the 24-h scale is the corticosterone produced by bisected adrenal glands incubated with KRb only, and the Mt content of the APH, in keeping with earlier work. Reactively (beta-) rhythmic is the response, again along the 24-hr scale, of the adrenal to Sy or Mt and the response of the anterior pituitary to Mt at different circadian times. Modulatory (gamma and delta) rhythms characterize the Mt effect on the (beta-rhythmic) adrenal stimulation by Sy. Chronomodulations thus emerge as more or less complete sequences of a decrease, no effect and/or increase; or, rather, as an attenuation, no effect and/or amplification by the pineal, of the pituitary effect upon the adrenal. Linear least-squares rhythmometry, mostly single cosinors, describes and quantifies circadian (alpha, beta, and gamma) and infradian (delta) rhythms in the original series and in the differences in responses [beta - alpha] and [gamma - beta] (P less than 0.001 in each case). These spontaneous alpha, response beta, modulatory gamma, and frequency-divided delta rhythms reveal a collateral neuroendocrine hierarchy, characterized by the pineal feedsideward phenomenon, as a feature of interactions recurring with circadian and infradian frequencies. PMID- 3037559 TI - Peculiarities of the endocrine time structure in noninsulin-dependent adult-onset (type II) diabetes mellitus. AB - Twenty noninsulin-dependent elderly diabetic patients, ten of whom were treated by oral hypoglycemic agents and ten of whom were regulated by diet alone, and 20 clinically healthy subjects matched for age, sex, height, and weight were examined with six blood and six urine samples at 4-hr intervals over a 24-hr span. Plasma ACTH, cortisol, aldosterone, and dehydroepiandrosterone-sulfate (DHEA-S) were determined by radioimmunoassay (RIA); epinephrine, norepinephrine, and dopamine in urine were determined by high-pressure liquid chromatography (HPLC); and magnesium in urine was determined colorimetrically on a DuPont ACA. There were a number of changes in some of these functions in type II diabetic patients with and without oral hypoglycemic agents that appear to be of interest. The circadian mean in plasma ACTH concentration in diabetic patients with and without oral hypoglycemic agents is significantly higher than in matched nondiabetic controls. The plasma aldosterone concentration is similar in type II diabetics treated by diet only and in matched controls but is statistically significantly elevated in patients on oral hypoglycemic agents. Correspondingly, the urinary excretion of sodium in type II diabetic patients on oral hypoglycemic agents is lower than in matched controls. The plasma cortisol concentration is unchanged in type II diabetic patients treated by diet alone but shows a slight increase in patients on oral hypoglycemic agents. The circadian means of plasma DHEA-S concentration is slightly higher in diabetic patients with and without oral hypoglycemic agents than in matched controls. This elevation, however, does not quite reach the 95% level of statistical significance. Urinary norepinephrine excretion in type II diabetic patients is similar to that in matched controls. The urinary epinephrine excretion in diabetics with and without oral hypoglycemic agents, however, was lower than in controls, and the urinary excretion of dopamine was higher in the diabetics. The urinary magnesium excretion in type II diabetic patients was lower than in matched controls. PMID- 3037560 TI - Immune mechanisms in Rous sarcoma regression. PMID- 3037561 TI - Allograft and antibody responses of 15I5-B congenic chickens. AB - Seven congenic lines of chickens that differ from the parental inbred line RPRL 15I5 for genes in the major histocompatibility (B) complex are under development. After 2 to 5 generations of crossing B-homozygous chickens were produced for interim histocompatibility and antibody competence tests. Chickens in each of the B-congenic lines had antigens determining rapid skin graft rejection and stimulation of mixed lymphocyte responses by 15I5 chickens. Several lines repeatedly developed different antibody responses after immunization with sheep red blood cells, killed Brucella abortus, and Infectious Bursal Disease Virus Vaccine. PMID- 3037562 TI - Immunization of chickens against Marek's disease with cell-free supernatant from the JMV-1 lymphoblastoid cell line. PMID- 3037563 TI - Mechanism of T cell immunosuppression by infectious bursal disease virus of chickens. AB - Spleen cells of chickens infected with IBDV responded poorly to in vitro stimulation with Con A. The mitogenic hyporesponsiveness was due to the presence of suppressor cells that could be removed by pretreatment of IS cells with carbonyl iron or cytodex-3 microcarrier beads. Addition of suppressor cells to normal spleen cells prevented the normal cells from responding to mitogen. Cell to-cell contact between responder and suppressor cells was not necessary for suppression to occur; the effect was mediated by soluble product(s) released by the suppressor cells. IS cells were also deficient in producing mitogen-induced IL-2 and addition of exogenous IL-2 did not restore mitogenic response of IS. PMID- 3037564 TI - Systemic and local antibody responses in chickens after infection and vaccination with infectious bronchitis virus. PMID- 3037565 TI - Chicken B lymphocyte differentiation: ontogeny, bursal microenvironment and effect of IBD virus. PMID- 3037566 TI - Effects of chronic ethanol exposure on adenylate cyclase activities in the rat. AB - The effects of chronic ethanol administration on striatal and cerebellar adenylate cyclase systems were investigated in the rat. The chronic ethanol treatment resulted in behavioral tolerance, but no difference in the sensitivity of adenylate cyclase to in vitro ethanol was observed. In one set of experiments using 173-195 g rats, GTP-, dopamine- and NaF-stimulated adenylate cyclase activities in the striatum were higher in rats chronically treated with ethanol when compared to animals pair-fed the liquid diet. However, no difference in adenylate cyclase activity was observed in cerebellar or striatal tissues when larger rats (280-385 g) were used. In conclusion, an adaptive change in activation of adenylate cyclase by in vitro ethanol does not occur after chronic ethanol treatment. The observed changes in enzyme activity measured in the absence of in vitro ethanol do not appear to be a simple, direct effect of chronic ethanol treatment. PMID- 3037567 TI - Delineation of mu-antagonist, partial kappa agonist and non-opioid agonist activity of cyclazocine using urinary output of rats. AB - The effects of cyclazocine (SC) were studied under a variety of test conditions to determine its various activities on urinary output. Cyclazocine antagonized the morphine-induced (20 mg/kg) antidiuretic effect in water-loaded (3 ml/100 g b.wt.) rats at low doses (0.08, 0.16 and 0.32 mg/kg). At higher doses (1.25, 5 and 20 mg/kg), cyclazocine caused diuresis in normally hydrated rats. The diuresis produced by 1.25 and 5, but not 20 mg/kg, was antagonized by naloxone (10 mg/kg). In water-deprived rats and in normally hydrated rats, cyclazocine, in a dose-related fashion, antagonized the diuretic effects of 0.08 mg/kg of bremazocine. These results are compatible with the interpretation that cyclazocine has mu-antagonist activity at low doses, then partial kappa-agonist activity at intermediate doses, followed by non-opioid agonist activities at the higher doses. PMID- 3037568 TI - The direct enhancement of positive palatability by chlordiazepoxide is antagonized by Ro 15-1788 and CGS 8216. AB - In a previous study, it was found that positive, palatability-dependent consummatory reactions in rats to intraorally infused tastes were facilitated by chlordiazepoxide (10 mg/kg). In contrast, the rats' more neutral or aversive reactions to these tastes were not facilitated by chlordiazepoxide. This suggested that chlordiazepoxide might selectively enhance the positive palatability of tastes. This effect was replicated in the present experiment, and in addition, the benzodiazepine antagonists Ro 15-1788 and CGS 8216 were found to counteract the enhancement of positive ingestive reactions produced by chlordiazepoxide. These antagonist effects generally suggest that the benzodiazepine receptor complex may be involved in making tastes more palatable after chlordiazepoxide administration. PMID- 3037569 TI - Opioid-induced linear running in obese (ob/ob) and lean mice. AB - Earlier research has shown that opioids stimulate behavioral activation in mice whereas opioid antagonists attenuate this activation. We conducted an experiment to determine the dose-response curve of FK33824, a potent Met-enkephalin analogue. FK33824 produced an unusual form of behavioral activation we called "linear running" in which the mice ran continuously in one direction and were nearly oblivious to environmental stimuli. This may be the kind of running that occurs naturally during migration. Wheel running activity of genetically obese (ob/ob) and lean (C57BL/6J ?/+) mice was measured following the intracerebroventricular infusion of 0.1, 1.0, 10.0 and 100.0 ng of FK33824. The lowest dose did not alter baseline running, whereas the 1.0 and 10.0 ng doses significantly increased running in both genotypes. We found a genotype difference with the highest dose tested, the lean mice ran at baseline levles and displayed ataxia whereas the obese mice continued to show increased running without ataxia. We hypothesize that genetic differences in the enkephalin mechanisms of C57 lean and obese mice are responsible for linear running. PMID- 3037570 TI - Clonidine and prazosin block the iminodipropionitrile (IDPN)-induced spasmodic dyskinetic syndrome in mice. AB - The effects of drugs with selective action on alpha 1- or alpha 2-adrenoceptors were investigated on persistent head twitches, vertical neck dyskinesia, and the random circling behaviors induced by chronic intraperitoneal injections of IDPN. The alpha agonist clonidine and the alpha antagonist prazosin inhibited the IDPN induced behavioral syndrome whereas the alpha antagonist yohimbine had no significant effect. These results suggest that dysregulation of a facilitatory noradrenergic input to cortical and/or subcortical motor areas may be involved in the abnormal movements caused by chronic treatment with IDPN. PMID- 3037571 TI - Anti-shock effect of ACTH-(1-24) in rats: comparison between intracerebroventricular and intravenous route of administration. AB - In an experimental model of hypovolemic shock in rats, produced by withdrawing about 50% of the estimated total blood volume, and causing 100% deaths within 30 min, ACTH-(1-24) dose-dependently improved arterial and pulse pressure and increased survival rate. The intracerebroventricular (i.c.v.) route of administration was more effective than the intravenous (i.v.) route: at the dose of 24 micrograms/kg, 45% and 91% of rats were still surviving 2 hr after i.v. and i.c.v. treatment, respectively. At higher doses of ACTH-(1-24), the survival rate rose to 100% regardless of the route of administration, but arterial pressure increased more after i.c.v. than after i.v. injection. These data suggest that the CNS plays an important role in the anti-shock effect of ACTH. PMID- 3037572 TI - Interaction of dopamine, (+/-)-dobutamine and the (-)-enantiomer of dobutamine with alpha- and beta-adrenoceptors in the pulmonary circulation of the dog. AB - The effects of dopamine, (+/-)-dobutamine (racemic mixture) and (-)-dobutamine on alpha-adrenoceptor-mediated vasoconstriction were evaluated in the pulmonary circulation of the anesthetized dog. The drugs were studied in the absence and presence of propranolol (1 mg/kg, i.v.) in order to assess beta-adrenoceptor mediated effects in the pulmonary circulation. Intra-arterial administration of dopamine, (+/-)-dobutamine and (-)-dobutamine elicited dose-dependent increases in pulmonary perfusion pressure, reflecting increases in pulmonary vascular resistance. In control animals, dopamine elicited the largest increases in pulmonary perfusion pressure (45% above resting pulmonary pressure) followed by ( )-dobutamine (30% increase) and (+/-)-dobutamine (15% increase). The pressor effects of dopamine in the pulmonary circulation were mediated by both postjunctional vascular alpha 1- and alpha 2-adrenoceptors, since prazosin, (100 micrograms/kg, i.v.), a selective alpha 1-adrenoceptor antagonist, and rauwolscine (100 micrograms/kg, i.v.), a selective alpha 2-adrenoceptor antagonist, both inhibited the vasopressor response elicited by dopamine to roughly equivalent degrees. Pulmonary vasoconstriction produced by (+/-) dobutamine and (-)-dobutamine was mediated primarily by postsynaptic vascular alpha 1-adrenoceptors, although alpha 2-adrenoceptor-mediated vasoconstriction was observed. In propranolol-pretreated animals, the increase in pulmonary perfusion pressure elicited by dopamine and (-)-dobutamine was qualitatively and quantitatively similar to that observed in control animals, suggesting that these agents have little activity on vascular beta 2-adrenoceptors in the pulmonary circulation. In marked contrast, the maximum pulmonary vasopressor response obtained with (+/-)-dobutamine were greater in propranolol-pretreated animals, indicating that (+/-)-dobutamine also has the capacity to stimulate pulmonary vascular beta 2-adrenoceptors which mediate pulmonary vasodilation that, in part, mask alpha-adrenoceptor-mediated pulmonary vasoconstriction. Since the (-) enantiomer of dobutamine has little or no beta 2-adrenoceptor agonist activity, the beta 2-adrenoceptor-mediated effect of (+/-)-dobutamine must result from the (+)-enantiomer as has been previously proposed. The results of the present study indicate that dopamine has a greater propensity for increasing pulmonary vascular resistance, and therefore pulmonary arterial blood pressure, relative to (+/-) dobutamine. This results, at least in part, from the relatively weaker activity of dopamine in stimulating pulmonary vascular beta 2-adrenoceptors which mediate vasodilation.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3037573 TI - DNA topoisomerase II: a primer on the enzyme and its unique role as a multidrug target in cancer chemotherapy. AB - Based on the weight of evidence accrued in the past eight years, there is little question that the nuclear enzyme, topoisomerase II, serves as a common intracellular target for the cytotoxic effect of drugs of widely varying structure. The enzyme appears to be unique as a chemotherapy target in that it is recruited into a lethal process under the influence of drug. Its role contrasts sharply with other more classical chemotherapy targets, such as dihydrofolate reductase, whose activity must be successfully inhibited for the expression of cytotoxicity. Resistance to inhibitors of this enzyme frequently results from marked elevations in intracellular enzyme content. In contrast, the presence of topoisomerase is required for drug effect, and, in general, the greater the cellular content of the enzyme, the more sensitive the cell will be to these agents. However, important issues remain unresolved. The biochemical events that are initiated by cleavable complex formation and result in cell death must be more fully defined. It is likely a better understanding of the drug-enzyme interaction will be required for rational drug development. Finally, those aspects of the drug-topoisomerase interaction that confer therapeutic selectivity and/or clinical resistance are of paramount importance if the phenomenon is ever to be fully exploited. PMID- 3037574 TI - Spectroscopic studies of cutaneous photosensitizing agents--X. A spin-trapping and direct electron spin resonance study of the photochemical pathways of daunomycin and adriamycin. PMID- 3037575 TI - Lipid peroxidation and other membrane damage produced in Escherichia coli K1060 by near-UV radiation and deuterium oxide. PMID- 3037576 TI - Malignant clear-cell hidradenoma. AB - In this paper the authors present a case of malignant clear-cell hidradenoma developed on the dorsum of the hand. This is only the eleventh complete report in the literature, including follow-up, and only the third to include necropsy. It is the first to find both lung and myocardial metastases. The tumor was very invasive and was treated first by wide surgical excision and radical node dissection, and when the tumor recurred, amputation of the arm was done. The world literature was carefully reviewed and compared with this case. Owing to the already known failure of radiotherapy, and since chemotherapy was not successful in our patient, the authors advise early aggressive surgery, including block dissection of the regional nodes, and thereafter a careful follow-up. PMID- 3037577 TI - [Diagnostic value of the H-reflex index in alcoholic polyneuropathy]. AB - We examined the H-reflex-index of triceps surae muscle (HI) and the motor nerve conduction velocity of the tibial nerve (mNLG) of 56 alcohol dependent patients. The results were compared with neurologic disorders, duration of the dependency and the type of alcoholism according to Jellinek. The HI showed significant more pathological changes than the mNLG. PMID- 3037578 TI - The ACTH stimulation test before and after clinical recovery from depression. AB - Excessive cortisol secretion after cosyntropin (adrenocorticotropic hormone; ACTH) infusion in some depressed patients has suggested the possibility that the adrenal cortex may have heightened responsiveness to ACTH, and that this may contribute, in part, to activation of the hypothalamic-pituitary-adrenocortical axis. We administered an ACTH test and dexamethasone suppression test (DST) to 32 patients before and after treatment. Maximal cortisol response to ACTH demonstrated a significant decrease after treatment in the subgroup of melancholic/DST nonsuppressors (p = 0.04). When the cumulative cortisol response (CCR) to ACTH was examined, the DST nonsuppressors had a greater CCR decrease than suppressors (p = 0.03), and the melancholics a greater decrease than nonmelancholics (p = 0.02). The melancholic/DST nonsuppressor subgroup had the largest CCR decrease after treatment (p = 0.03), and these patients may represent a group of depressives with altered adrenocortical function that tends to "normalize" with clinical recovery. PMID- 3037579 TI - Pituitary adrenocortical unresponsiveness in lactate-induced panic. AB - The hypothalamic-pituitary adrenal (HPA) axis responds to a variety of physical and emotional stimuli with increased output of adrenocorticotropic hormone (ACTH) and cortisol, yet there is little known about the activity of this system during episodes of severe anxiety in patients with DSM-III-defined anxiety disorders. To explore further whether alterations of the HPA axis occur during various anxiety states, we measured ACTH and cortisol during lactate infusion in patients with panic disorder and agoraphobia. In eight patients who panicked during lactate infusion, there were no elevations in either ACTH or cortisol. Further, the patterns of hormone secretion did not differ among patients who panicked, nonpanicking patients, or controls. This negative result suggests that the neurobiological mechanisms that mediate panic differ from those responsible for other fear responses. PMID- 3037580 TI - Differential 3H-imipramine platelet binding in patients with panic disorder and depression. AB - 3H-Imipramine (3H-IMI) binding to platelet membranes was measured in 19 patients with agoraphobia with panic attacks, 9 patients with major depression, and 22 healthy subjects. In comparison to healthy subjects, the maximal number of binding sites (Bmax) was significantly decreased in depressed patients but not in panic disorder patients, and the apparent affinity of binding was slightly decreased in depressed patients but not in panic disorder patients. The Bmax and Kd of 3H-IMI platelet binding did not differ between panic disorder patients with and without a history of a major depressive episode. Thus, 3H-IMI platelet binding is clearly different in patients with panic disorder compared to those with an active depression. Because 3H-IMI binding is associated with the serotonin reuptake site in platelet and brain membranes, these findings give further support to abnormalities in serotonergic function in patients with major depression. PMID- 3037581 TI - Hypertension in depression. PMID- 3037582 TI - Selective effects of ECT on hypothalamic-pituitary activity. AB - The hypothesis that ECT produces selective effects on hypothalamic-pituitary activity was investigated by determining the effect of ECT on pituitary hormone release in nine depressed patients. After ECT there were massive and rapid increases in the plasma concentrations of nicotine- and oestrogen-stimulated neurophysin (NSN and ESN), prolactin (PRL) and adrenocorticotropin (ACTH), smaller increases in plasma luteinizing hormone (LH) and cortisol, a significant decrease in plasma growth hormone (GH) concentration but no change in plasma thyrotropin (TSH). There was significant attenuation of PRL responses with repeated ECT. The hormonal responses to ECT cannot simply be attributed to stress, since a similar pattern of increases in plasma hormone concentrations did not occur in psychologically normal patients in whom plasma hormone concentrations were measured during induction of anaesthesia and abdominal incision for cholecystectomy. Analysis of these hormonal responses in terms of the knowledge available on the neurotransmitter control of pituitary hormone release suggests that some of these hormonal responses to ECT may be mediated by the activation of serotonergic neurones, while others are probably due to direct stimulation of the neuroendocrine neurones themselves. PMID- 3037583 TI - Hypothalamic-pituitary-adrenal-cortical activity in anorexia nervosa and bulimia. AB - We studied hypothalamic-pituitary-adrenal-cortical (HPA) activity in nine underweight women with anorexia nervosa, 12 women of normal body weight with bulimia, and nine control subjects. The measures of HPA activity were the pattern of plasma cortisol secretion over 24 hr and the responses of plasma cortisol to dexamethasone suppression and to low dose ACTH stimulation. The patients with anorexia nervosa had significantly elevated 24 hr concentrations of plasma cortisol compared to the controls and showed significantly less cortisol suppression following dexamethasone. There was no difference between patients with anorexia nervosa and controls in the rise in plasma cortisol following ACTH. On most measures of HPA activity, the normal weight patients with bulimia were indistinguishable from the controls. These results suggest that HPA activity is normal in most patients of normal body weight with bulimia and that the psychological and behavioral disturbances common to both anorexia nervosa and bulimia are, in the absence of significant weight loss, insufficient to produce major alterations in HPA activity. PMID- 3037584 TI - Feedback action and tonic influence of corticosteroids on brain function: a concept arising from the heterogeneity of brain receptor systems. AB - Two types of corticosteroid receptors can be distinguished in rat brain. The type 1 receptor resembles the kidney mineralocorticoid receptor and has two functional expressions in brain, i.e. type 1 corticosterone (CORT) preferring sites (CR) and mineralocorticoid receptors (MR). The type 2 receptor is similar to the liver glucocorticoid receptor (GR). CORT binds to both CR and GR. The localization, binding specificity, and capacity of the receptor systems have served as criteria to evaluate steroid dependent events in brain biochemistry and behaviour. The GR is widely distributed in neurons and glial cells, with the highest density in frontal brain regions. The GR becomes occupied concomitant with rising plasma CORT levels after stress and as part of the circadian rhythm. The GR mediates the feedback action of CORT on stress-activated brain processes. The CR has its predominant localization in neurons of the septo-hippocampal complex and has a ten-fold higher affinity for CORT than that of the GR. The CR is, at all times of intact adrenocortical secretion, 90% or more occupied by endogenous hormone. The CR mediates a tonic influence exerted with stringent specificity by CORT on hippocampus-associated functions, e.g. cognition, mood, and affect. CORT, via the CR, thus contributes to hippocampus function in interpretation of sensory information, leading to appropriate neuroendocrine and behavioural responses, which are themselves subsequently subject to feedback action via the GR. The MR mediates the mineralocorticoid effect on salt and water balance and its behavioural corollary of salt appetite. The anatomical localization of the MR system is as yet ill-defined, although functional studies suggest circumventricular organs as mineralocorticoid target sites. The CR and the MR have in common the high affinity for mineralocorticoids, but the CR is defined by its exclusive responsiveness to CORT as its agonist. The CR and MR probably represent the same chemical receptor modality (type 1), which is expressed differentially depending on the presence of extravascular corticosteroid binding globulin (CBG) in the vicinity of the receptor. GR capacity is subject to autoregulation. Chronic stress, senescence, and chronic CORT administration reduce GR number, with, as a consequence, a less efficient feedback signal. The CR number seems not to be under the control of corticosteroids, probably since the receptor sites are extensively occupied by endogenous hormones. The CR number displays a circadian rhythm and is reduced during senescence.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3037585 TI - A quantitative cerebral and whole body autoradiographic study of a intravenously administered benzodiazepine antagonist 3H-Ro 15-1788 in mice. AB - 3H-Ro 15-1788, a benzodiazepine receptor antagonist, was injected IV into male and pregnant mice. Autoradiograms were prepared from sagittal sections of animals killed after 30 s to 48 h. In the adult animal there was a rapid and high initial accumulation of radioactivity in the brain as compared to other organs and tissues. The highest accumulation was found in cortical brain areas, such as the olfactory bulb and the frontal cortex. Cerebellar cortex, globus pallidus, amygdala, substantia nigra, colliculus, hippocampus and pons followed in rank order. The rate of decline of radioactivity was highest in the pons and lowest in the olfactory bulb. The initial disappearance of radioactivity from the cerebellum was higher than from the other brain regions. Ro 15-1788 was rapidly eliminated; 4 h after drug administration there was an almost complete clearance of radioactivity from all tissues. After 24 h no trace of activity remained in the animal. The distribution of radioactivity at later time points indicates that metabolites of Ro 15-1788 are eliminated by fecal, urinary and nasal secretion. In the fetus also there was an early accumulation of radioactivity in the central nervous system. The radioactivity in fetal organs was lower than in the mother at all time intervals. PMID- 3037586 TI - [Root alloplasty for auto-alloplastic replantation--a new method of surgical tooth restoration (preliminary report)]. PMID- 3037587 TI - Pulmonary endothelial dysfunction induced by unilateral as compared to bilateral thoracic irradiation in rats. AB - Rats were sacrificed 2 months after a single dose of 10-30 Gy of 60Co gamma rays delivered to either a right unilateral or a bilateral thoracic port. Four indices of lung endothelial function were measured: the activities of angiotensin converting enzyme (ACE) and plasminogen activator (PLA) and the production of prostacyclin (PGI2) and thromboxane (TXA2). The number of macrophages recovered by bronchoalveolar lavage (BAL) and the degree of right ventricular hypertrophy (an index of pulmonary hypertension) also were determined. Right lung ACE and PLA activity decreased linearly, and PGI2 and TXA2 production increased linearly with increasing radiation dose. The response curves for right unilateral and bilateral thoracic irradiation were not significantly different. In contrast, bilateral irradiation was more toxic than unilateral, since rats exposed to the former exhibited decreased body weight, an increased incidence of pleural effusions, an increase in the number of macrophages recovered by BAL, and right ventricular hypertrophy. These data demonstrate that pulmonary endothelial dysfunction induced by hemithorax irradiation represents a direct response of the endothelium to radiation injury and is not secondary to other phenomena such as shunting of function to the shielded lung. PMID- 3037588 TI - Modifications in repair and expression of potentially lethal damage (alpha-PLD) as measured by delayed plating or treatment with beta-araA in plateau-phase Ehrlich ascites tumor cells after exposure to charged particles of various specific energies. AB - The ability of Ehrlich ascites tumor cells (EAT cells) to repair potentially lethal damage (alpha-PLD) as demonstrated by either an increase in survival after delayed plating or a decrease in survival after treatment with beta arabinofuranosyladenine (beta-araA) was investigated after exposure to protons, deuterons, 3He, 4He, and heavy ions of various specific energies. A significant amount of repair or fixation was observed after delayed plating or treatment with beta-araA, respectively, in cells that were exposed to protons of 6-21 MeV energy, reflecting mainly variations in the survival curve shoulder width. Four hour treatment with 80 microM/liter beta-araA resulted in an exponential survival curve for all proton energies tested. A decrease in particle energy increased killing and caused a reduction in Dq without a significant change in D0. The survival curve obtained after exposure of cells to 3.4 MeV protons had only a small shoulder and was only slightly modified by either delayed plating or treatment with beta-araA, suggesting a decrease in the induction rate of alpha PLD. Similar results were also obtained after exposure to deuterons and 4He ions. The results are interpreted as indicating the importance of the specific particle energy and the delta-electron spectrum in the induction of alpha-PLD. When the results of delayed plating of cells exposed to protons, deuterons, or helium ions were pooled, an exponential relationship between Dq and penumbra radius was indicated. After exposure to 40Ar ions of 18 MeV specific energy, a shouldered survival curve was obtained, and beta-araA significantly enhanced killing by modifying Dq as well as D0, a result that also suggests induction of repairable damage by the delta particles produced and interaction of lesions induced within the core of the ion path with penumbra lesions. Based on these results a model is proposed assuming that alpha-PLD results from interaction, during the course of repair, of pairs of DNA lesions induced within a distance di. The model assumes the existence of a critical separation distance dic, with the property that pairs of lesions induced with separation distance shorter than dic (expressed as number of base pairs) will always be expressed as lethal, and the existence of a maximum separation distance dim, with the property that pairs of lesions induced with separation distance larger than dim will not interact.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3037589 TI - [Magnification mammography using 0.1 mm microfocus. Comparison of grid and spot view magnification]. AB - Phantom measurements of resolution and dosimetry of 0.1 mm microfocus spot magnification mammography and 0.3 mm grid technique using different film-screen systems form the basis of this study. The diagnostic efficiency of spot views with these methods is evaluated according to the detection and analysis of microcalcifications, and the assessment of tumor margins. PMID- 3037590 TI - Conventional and rapid MR imaging of the liver with Gd-DTPA. AB - Twenty-three patients with malignant hepatic tumors underwent magnetic resonance (MR) imaging before and after intravenous administration of gadolinium-diethylene triaminepentaacetic acid (DTPA). Two different doses were used, 0.1 mmol/kg and 0.2 mmol/kg. The larger dose proved to be more effective than the smaller dose. The signal-enhancement-to-noise ratio was significantly larger in the tumor than in the liver (2 alpha less than or equal to .05). In a moderately T1-weighted spin echo (SE) sequence (SE 400/30) (repetition time [TR] msec/echo time [TE] msec), the tumor was better defined 6 minutes after administration of Gd-DTPA. More strongly T1-weighted sequences--that is, SE 200/20 and inversion recovery 1,500/35/400 (TR msec/TE msec/inversion time, msec)--showed significantly worse contrast between tumor and liver (signal-difference-to-noise ratio [SD/N]) 10 and 15 minutes after administration (2 alpha less than or equal to .05). On the other hand, the low SD/N in the rapid MR imaging sequence was significantly improved (2 alpha less than or equal to .05). The most important indications for administration of Gd-DTPA in diagnosing hepatic tumors are the presentation of perfusion conditions and contrast optimization in rapid MR images. PMID- 3037591 TI - Cavernous hemangioma of the liver: detection with single-photon emission computed tomography. AB - The roles of single-photon emission computed tomography (SPECT) and planar imaging with technetium-99m-labeled red blood cells in the diagnosis of cavernous hemangioma of the liver were evaluated. The study group consisted of 26 consecutive patients referred for evaluation of liver lesions. A total of 23 cavernous hemangiomas were found, all of which showed decreased or normal flow and delayed uptake of the radiotracer. SPECT demonstrated 13 hemangiomas that were not detected with planar imaging; both modalities demonstrated the other ten lesions. Lesions that were not cavernous hemangiomas showed either normal (n = 6) or increased (n = 4) flow; none had delayed increased uptake on either planar or SPECT images. SPECT with labeled red blood cells is an accurate method for the detection of cavernous hemangiomas of the liver and is more sensitive than planar imaging in depicting small lesions. PMID- 3037592 TI - Mammographic findings after breast cancer treatment with local excision and definitive irradiation. AB - Following local excision and definitive irradiation of 163 breast cancers in 160 women, alterations in mammographic patterns were observed for up to 7 years. Skin thickening was observed in 96% of mammograms obtained within 1 year of completing therapy and was most pronounced in women treated with iridium implant, chemotherapy, or axillary dissection. In 76% of mammograms, alterations in the parenchymal pattern, including coarsening of stroma and increased breast density, were seen at 1 year. Neither skin nor parenchymal changes progressed after 1 year. Within 3 years of treatment the parenchymal density, which usually regressed, did not change in all patients. At 3 years skin thickness and the parenchymal pattern had returned to normal in less than 50% of the breasts of these women. Scars developed in approximately one-quarter of women. They were present on the initial post-treatment mammogram and remained unchanged on serial studies. Coarse, benign calcifications also developed in the breasts of about one quarter of women. Microcalcifications developed in 11 breasts; biopsy specimens of six were benign. Benign microcalcifications may be related to therapy. PMID- 3037593 TI - Distribution of iodized oil within the liver after hepatic arterial injection. PMID- 3037594 TI - [Occlusion and prosthetics]. PMID- 3037595 TI - [Prevention of osteoporosis]. PMID- 3037596 TI - [Evaluation of crowns after impressions with a combination of reversible and irreversible hydrocolloids]. PMID- 3037597 TI - [Freedom in centric. A practical occlusion concept]. PMID- 3037598 TI - [Differential diagnosis of facial pain]. PMID- 3037599 TI - [Gold band prosthetics in an Etruscan technic. A new discovery in West Anatolia]. PMID- 3037600 TI - [Periodontology--yesterday, today, tomorrow]. PMID- 3037601 TI - [Location of crown margins from the periodontal viewpoint]. PMID- 3037602 TI - [Psychosomatic problems in dental practice]. PMID- 3037603 TI - [The corrosion process in the mouth]. PMID- 3037604 TI - [Mandibular movement and its clinical significance]. PMID- 3037605 TI - [Arthrosis models of the temporomandibular joint]. PMID- 3037606 TI - Circadian rhythms in mammalian neurotransmitter receptors. AB - At the present time, the following summary statements can be made as to 24-hour changes in receptor binding. In all receptors studied in homogenates from whole rat forebrain (alpha 1, alpha 2, beta-adrenergic, muscarinic cholinergic, dopaminergic, 5HT-1, 5HT-2, adenosine, opiate, benzodiazepine, GABA, imipramine), significant variations over 24 hours have been documented. The receptor rhythms measured change in wave form, amplitude, and phase throughout the year, even though the animals have been kept on a defined and constant LD cycle. Whether these rhythms are truly seasonal requires further investigation. The rhythms are circadian: i.e. they persist in the absence of time cues, and the unimodal rhythms do not persist after lesion of the putative circadian pacemaker in the suprachiasmatic nuclei. The rhythms can be uni- or bimodal, and each brain region shows a particular pattern. The pattern can be different for the same ligand in different nuclei of a given brain region (e.g. hypothalamus). Nearly all studies of receptor rhythms have been carried out in rats; the results vary according to strain and even within the same strain from different breeding lines. Receptor rhythm characteristics are modified by age: e.g. the amplitude, phase, as well as the 24-hour mean of binding to a given ligand in a defined brain region. The changes in number of binding sites over 24 hours can be correlated with amine turnover, second messenger, or function of that brain region; however these relationships, although consistent within a region, do not hold for all regions. If gradual changes in CNS neurotransmitter receptor function are considered important in the pathogenesis of schizophrenia and affective disorders and the mode of action of psychopharmacological agents, then consideration of the short term rapid change over 24 hours is equally necessary. Chronic treatment with a number of psychoactive drugs known to induce up- or down-regulation of receptor number, also induces marked changes in circadian rhythm parameters of wave form, amplitude, phase and 24-hour mean. This is of methodological importance for single time-point studies, since the interpretation of the results will depend on time of day. Preliminary evidence supports the assumption that the significant variation in receptor binding throughout the day may underlie the well-known circadian rhythms of susceptibility to many CNS drugs. New findings of circadian rhythms in receptors on blood cells indicate the relevance of these changes also in human physiology. PMID- 3037607 TI - Differential effects of leukotrienes B4 and C4 on bovine aortic endothelial cell proliferation in vitro. AB - The effect of leukotrienes derivated from arachidonic acid was studied on vascular endothelium proliferation. The peptido-leukotriene LTC4 (0.1 nM - 0.1 microM) promoted a dose-dependent growth of bovine aortic endothelial cells in culture with a maximal effect at 10 nM. This proliferative activity could be receptor-mediated since LTC4 specifically bound to endothelial cell membranes with a Kd value of 50 nM. The leukotriene B4 did not induce any significant proliferation in the same range of concentrations. This result was consistent with the lack of LTB4 specific binding sites. This data suggests that LTC4 could be one of the factors implicated in angiogenesis during inflammatory processes. PMID- 3037608 TI - Biosynthesis, characterization and inhibition of leukotriene B4 in human whole blood. AB - We have evaluated the biosynthesis, characterization and inhibition of Leukotriene (LT) B4 in unstimulated and in A23187-stimulated human whole blood. LTB4 was assayed by radioimmunoassay (RIA) both in unextracted serum and after extraction and thin-layer chromatography (TLC). Unstimulated human whole blood allowed to clot at 37 degrees C for 60 min produced only trace amounts of LTB4 (0.16 +/- 0.05 ng/ml, mean +/- SD, n = 3). LTB4-like immunoreactivity (ir-LTB4) detectable in unstimulated serum samples was largely overestimated by direct RIA, most likely because of interfering substance(s) unrelated to cyclooxygenase or lipoxygenase activity. Incubation of human whole blood with A23187 (2-10 microM) resulted in a concentration-dependent stimulation of LTB4 production. At 10 microM A23187, ir-LTB4 was 18 +/- 2.4 ng/ml (mean +/- SEM, n = 28). In A23187 stimulated serum samples, LTB4 concentrations measured by direct RIA correlated in a statistically significant fashion with those measured after extraction and TLC. Nafazatrom added in vitro caused a dose-dependent inhibition of A23187 stimulated ir-LTB4 production with an IC50 of 17 microM. PMID- 3037609 TI - Toward an understanding of cortisol dysregulation in major depression: a review of studies of the dexamethasone suppression test and urinary free-cortisol. PMID- 3037610 TI - The correspondence of plasma ACTH and cortisol before and after dexamethasone in healthy and depressed subjects. PMID- 3037611 TI - Corticotropin releasing hormone: clinical endocrinology and implications for Cushing's disease and endogenous depression. PMID- 3037612 TI - Etiologic implications of corticosteroid changes in affective disorder. PMID- 3037613 TI - Insulin tolerance test in depression. PMID- 3037614 TI - The ACTH stimulation test in depression. PMID- 3037615 TI - Disappearance of Tc-99m HMDP tumor uptake in oat cell carcinoma after irradiation and chemotherapy. AB - Extraosseous localization of bone-seeking radiopharmaceuticals has been reported in various tumors, presumably on the basis of active calcium deposition in the tumors. We report a case of oat cell carcinoma in which the initial localization of Tc-99m-HMDP in the tumor disappeared after irradiation and chemotherapy. The disappearance of tracer uptake coincided with regression of the mass as seen in the chest radiograph. This finding may have potential application in determining tumor response to anticancer therapy. PMID- 3037616 TI - Long-term results of treatment with mild toxicities in small cell lung carcinoma. AB - Results of treating small cell lung carcinoma with mild toxicities were investigated retrospectively. From 1974 to 1981, 38 patients were treated according to three institutional protocols: radiation alone (RAD), radiation with ifosfamide (IFOS), and radiation with adriamycin, vincristine, and cyclophosphamide (AOC). Of the three five-year survivors, two were treated with IFOS and one with RAD. The time period between admission and the beginning of radiotherapy was longer and the treatment field smaller in AOC than in the other two protocols. IFOS and RAD showed less toxicity. Our study indicated that IFOS and RAD should be regarded as curative treatments for limited disease and are suitable for patients who do not have sufficient bone marrow reserve. PMID- 3037617 TI - Survival and quality of life after continuous accelerated radiotherapy of glioblastomas. AB - One hundred and thirty three patients were given radiotherapy after subtotal resection of glioblastoma with different total doses and fractionation schedules including 38 patients receiving continuous accelerated fractionation with 3 fractions of 1.6 Gy per day up to 60 Gy in 2 weeks. Tolerance of the accelerated schedule was as good as of the conventional schedules but overall survival was not improved. PMID- 3037618 TI - Familial gastrointestinal polyposis: current considerations. PMID- 3037619 TI - Role of ultrasonography in the study of multilocular cystic nephroma. PMID- 3037620 TI - Scintigraphic diagnosis of hemorrhagic gastritis in a child. PMID- 3037621 TI - Effect of indomethacin and propranolol on the blood pressure and renin response to atriopeptin III in conscious rats. AB - The effect of prostaglandin synthesis inhibition and of beta-adrenoceptor blockade on the blood pressure and renin response to the synthetic atrial natriuretic peptide atriopeptin III was assessed in unanesthetized normotensive rats. This peptide was infused i.v. for 30 min at a rate of 1 microgram/min in rats pretreated either with indomethacin (5 mg i.v.) or propranolol (1 mg i.v.). The blood pressure reducing effect of atriopeptin III was attenuated neither by indomethacin nor by propranolol. Atriopeptin III per se did not modify plasma renin activity. Both the administration of indomethacin and of propranolol had a suppressing effect on renin release during atriopeptin III infusion. These data suggest that the vasodilating properties of atrial natriuretic peptides do not depend in the conscious normotensive rats on the production of prostaglandins. They also provide evidence that during infusion of such peptides, both prostaglandins and beta-adrenergic mechanisms are still involved in the regulation of renin secretion. PMID- 3037622 TI - Effect of vasoactive intestinal polypeptide on the cyclic adenosine monophosphate generating system in rat kidney glomeruli and cortical tubules. AB - The effect of in vitro addition of vasoactive intestinal polypeptide (VIP) on the 3'-5'-cyclic adenosine monophosphate (cAMP) generating system in rat kidney glomeruli and cortical tubules was studied. VIP did not stimulate cAMP accumulation in glomeruli; but VIP did stimulate, specifically and in a dose dependent manner, cAMP concentrations in tubular membranes with the addition of GTP. These results are consistent with the existence of a functionally active VIP receptor coupled to adenylate cyclase in rat kidney cortical tubules. PMID- 3037623 TI - [Combined chemotherapeutic and radiotherapeutic treatment of limited pulmonary microcytoma]. AB - Between 1978 and 1983, 34 patients (32 evaluable) suffering from limited small cell lung carcinoma (SCLC-L) were treated following the protocol polychemotherapy (CAV) plus thoracic cobalt teletherapy and "precautionary" cranial irradiation (30 Gy in 2 weeks). Minimum follow-up was 30 months. After induction chemotherapy there was complete remission (CR) in 20% of cases whereas at the end of induction chemo-radiotherapy there was complete remission (CR) in 44% (p less than 0.05) of cases. Median duration of the responds was 12 months. Total median survival is 15 months, median NED survival 32 months (6-90 months). Seven out of 14 CR patients received consolidated thoracic radiotherapy (Rt); 6 of these survived disease free for over 2 years. No salvage therapy carried out has proved useful. Only in one patient (3%) brain metastasis occurred. Iatrogenic toxicity was also kept within limits of brain level. The role Rt plays in increasing the CR percentage, in drastically diminishing the incidence of brain metastasis, in improving the quality of life by increasing the disease-free interval must be emphasized. Finally it should be noted that only CR patients have the possibility to become "long survivors". PMID- 3037624 TI - [The diabetic diet]. PMID- 3037625 TI - [Mental neuropathy associated with pulmonary small cell carcinoma]. PMID- 3037626 TI - [Study of the response of c-AMP, glucose and free fatty acids to stimulus with glucagon in rats in experimental hepatic cirrhosis]. PMID- 3037627 TI - [Clinical applications of the study of neuroendocrinologic rhythms]. PMID- 3037628 TI - [Meniscus imaging in MR tomography using surface coils. Anatomy, areas of degeneration, laceration formation]. AB - High resolution MR imaging using surface coils and thin-section techniques provides specific information when evaluating meniscal diseases. Extension and localisation of meniscal tears can be demonstrated as well as meniscal attachment areas. Diagnosis of degenerative changes is possible. We examined 7 healthy volunteers and 58 patients with suspected knee injuries. This study was done to discuss the use of MRI in the evaluation of meniscal diseases. PMID- 3037629 TI - [Sonographic study of Beclard's nucleus as a method of determining the maturity of the newborn infant]. AB - The size of the distal femoral epiphysis is an important criterion for determining the maturity of a neonate. The distal femoral epiphysis has been demonstrated and measured sonographically in two planes during the first month of life in 174 neonates. The sum of the epiphyseal diameters has been related to gestational age. The average sum of the sagittal and transverse diameter of both distal femoral epiphysis was 10.9 mm in the 35th week (standard deviation 7.6 mm) and increases to 38.4 mm at 43 weeks (standard deviation 6.3 mm). Linear regression of the mean values of the sum of the epiphyseal diameters y against gestational age x shows a correlation coefficient of 0.95 (regression gradient y = -117.7 + 3.7 x). Sonographic demonstration and measurements of the distal femoral epiphyses is a valid alternative to radiological examination. PMID- 3037630 TI - [Computerized tomography diagnosis of acetabulum fractures]. AB - One hundred and ten acetabular fractures in 102 patients were examined by CT. In 78% there was agreement with the findings on conventional radiological examination. CT altered the classification of fractures, particularly if these were complex. CT was superior to conventional radiology for demonstrating the course of the fractures and for identifying local complicating factors. In 68 surgical explorations, specificity of CT was found to be 97%. After the age of 40 there was a tendency for complex fractures. Analysis of the fractures in three dimensions often determines the need for surgical treatment, particularly if the weight-bearing portion of the acetabular roof and the posterior bony column has been fractured. PMID- 3037631 TI - [Skeletal involvement in malignant lymphoma]. AB - Of 114 patients with Hodgkin's disease reviewed, 13 (11%) had bone involvement. The typical osseous lesion in Hodgkin's disease is a secondary manifestation in the axial skeleton. The lesions are mainly lytic; there is no relation between the site of the affection and the character of the lesion. Of 261 patients with non-Hodgkin lymphoma, 19 (7.3%) had bone involvement. Non-Hodgkin lymphoma very rarely affects the bone primarily; these lesions are lytic. Involvement of the appendicular skeleton is reported to by typical; this cannot be confirmed by our study. Most secondary osseous lesions in non-Hodgkin lymphoma are found in the cranium, the spine and the sternum; this pattern of distribution as well as the mainly lytic appearance can be considered typical. PMID- 3037632 TI - CT detection and location of intraorbital foreign bodies. Experiments with wood and glass. AB - The series comprises 27 patients examined by CT to detect, locate or exclude a foreign body. 22 of them actually had an orbital foreign body. In three cases CT was the primary method and showed the foreign body correctly, while in 18 it was first detected in plain films and CT was performed to locate it. 20 metallic foreign bodies were hyperdense in appearance. Two cases had wooden foreign bodies, one with a density of +10 HU and the other hypodense with a value of about -434 to -446 HU. The latter piece of wood was first interpreted falsely as a bubble of gas. The results proved that the detection of metal is easy, but differentiation between wood and gas is problematical. Experiments conducted to determine the CT densities of different pieces of wood gave results varying from 618 HU to +23 HU. The highest densities obtained for glass varied from +522 HU to +2000 HU. The density of a plastic lens was -105 HU. PMID- 3037633 TI - [Digital subtraction dacryocystography and sialography]. AB - Fourteen examinations of the tear ducts and 21 sialograms were performed using digital subtraction and conventional techniques. In nine of the former and 15 of the latter, digital subtraction produced better results, in the remainder conventional technique was equally good. The advantages of digital subtraction are due to the removal of superimposed structures of the skull. Dose measurements at the level of the lens of the eye showed a greatly reduced radiation dose when using digital subtraction, as compared with conventional dacryocystography. Digital subtraction techniques with pulsed radiation and the ability to perform pixel shift is an optimal method for performing radiological examinations of the tear ducts and salivary glands. PMID- 3037634 TI - [Computerized tomography of the thorax in intensive care patients]. AB - Thoracic CT for patients in intensive care is cumbersome but provides important additional information in the presence of complicated lung changes. Total opacification of a lung field visualised on conventional portable films may be due to infiltration and/or fluid and/or collapse by using the clinical information in conjunction with densitometric measurements. CT may help in the differentiation of pulmonary oedema, particularly in the presence of ARDS and its complications. It is also possible to accurately localise abscesses and empyemas in the presence of extensive consolidation. This makes it possible to drain abscesses or empyemas, or pleural fluid in unusual situations, which has become loculated, or to aspirate a pneumothorax. PMID- 3037635 TI - [Computerized tomography imaging of the anterior cardiophrenic angle]. AB - The anterior cardiophrenic angle (ACA) is the lowermost anterolateral portion of the mediastinum. It is limited inferiorly by the diaphragm, anteriorly by the chest wall, posteriorly by the pericardium, and laterally by the parietal pleura. In 20% computed tomography (CT) of the normal ACA shows the diaphragm as a linear structure, and in 15% as a pseudotumorous structure that has to be differentiated from real tumours. The inferior sternopericardial ligament appears as a linear structure in 22%. In 3% nodular structures of up to 8 mm of diameter are visible in the normal ACA, probably corresponding to cardiophrenic angle nodes. These findings suggest that, exceptionally, normal cardiophrenic angle nodes may be demonstrated by CT. PMID- 3037636 TI - [Diagnosis of sarcoidosis using MR tomography]. AB - One hundred and thirty-three patients with lymph node enlargement from various causes were examined by MR tomography, using 0.5 and 1.5 Tesla. Amongst these, there were 27 patients with chronically enlarged lymph nodes and histologically confirmed sarcoidosis; these showed characteristic T1 intervals. Enlarged lymph nodes due to chronic inflammatory conditions can be differentiated from lymph nodes with malignant lymphoma, or from lymph node metastases from carcinomas. Although there are problems with the determination of relaxation times on MR tomography, this may be a valuable method for the differential diagnosis of sarcoid. PMID- 3037637 TI - [Nuclear spin tomography and computerized tomography in malignant hypernephroma]. AB - Forty patients with histologically confirmed hypernephromas were examined by MR and CT. As regards T-staging, MR had an accuracy of 97% and CT of 91%; for N staging, MR had an accuracy of 85% and CT of 91% and, with regard to infiltration of the renal vein, MR was correct in 88% of cases and CT in 81%. MR was most accurate in the pre-operative assessment of tumour spread, demonstrating caval involvement in 100% and lymph node metastases in 97%. On the other hand, infiltration of the renal pelvis could be assessed in only 65% of cases; in 27% the assessment was not possible and in 8% one could not be certain. CT proved to be at its best for staging of lymph nodes, but in 15% it was not possible to identify infiltration of the renal vein. MR was not greatly superior to CT in the preoperative diagnosis and staging of hypernephromas. PMID- 3037638 TI - [CT-guided sclerosing therapy of kidney cysts. Report of experience with 100 patients]. AB - Due to its diagnostic accuracy as well as safe and accurate puncture technique CT can be used advantageously for percutaneous sclerosing therapy of renal cysts. CT guided sclerosing therapy with pantopaque emulsions was carried out in 100 patients on an outpatient basis with altogether 107 solitary renal cysts. There were indications in all cases. Continuous follow-up of cyst involution in 96 patients with 103 cysts pointed out a reliable sclerosing effect in 95.1% of them. Major complications were neither registered in the puncture technique nor in the late urographic examinations. PMID- 3037639 TI - [Functional assessment, using micturition cystourethrography, of the unstable bladder in childhood]. AB - The accuracy and reliability of a modified method of micturating urethrocystography for the diagnosis of uncontrolled detrusor contraction in unstable bladders (sphincter-detrusor dyssynergy during bladder filling) was studied in 100 patients aged between four and 15 years. A comparison with a urodynamic method showed an accuracy of 92%, sensitivity of 93% and specificity of 90.7%. The technique and indications are described and the clinical relevance of the results is discussed. PMID- 3037640 TI - [Imaging diagnosis of ovarian tumors. Magnetic resonance tomography in comparison with computerized tomography and sonography]. AB - The present paper compares the results of MRT with sonography in 64 patients with tumours of the adnexa in 35 patients examined by CT. There was no difference between these three imaging methods as regards lateralisation of the lesion. MRT provided better differentiation because of the excellent demonstration of the uterus and of tumours of the adnexa. Detailed tissue characterisation, particularly as regards cystic lesions, provides improved diagnostic information. MRT has problems, however, because of its low spatial resolution and the difficulty in differentiation from bowel loops. At present sonography and CT is better at establishing a differential diagnosis. CT remains the method of choice for tumour staging. PMID- 3037641 TI - [Roentgenological demonstration of pathological changes of the mucosa of the small intestine]. AB - Use of a special high molecular guar preparation in small bowel contrast studies results in a roentgenological detection of the villous architecture of the small intestine. A diffuse villous atrophy results in a loss of the normal granular pattern. Segmental loss of granular mucosal texture is found in chronic radiogenic enteritis. In active Crohn's disease a coarser diffusely distributed granular appearance of the mucous surface occurs in addition to the segmental inflammatory changes. A coarse granular mucosal pattern with only segmental involvement is seen in mild malabsorption syndrome in the upper jejunum. Radiological signs for an intestinal protein loss syndrome is a coarse and weakly contrasted granular mucosal pattern in the lower ileum with moderate thickening of the jejunal folds and increased liquid content of the bowel. PMID- 3037642 TI - The incidence of hepatic hamartomas in tuberous sclerosis. Evaluation by ultrasonography. AB - Our series of tuberous sclerosis patients consisted of 23 children between 6 and 16 years of age and of 13 patients between 16 and 48 years of age. In the former group the incidence of multiple hepatic haemangiomas, estimated by grey-scale ultrasonography, is 13%, whereas this incidence is 23% in the group of older patients. The sign might be important for genetic counselling in formes frustes. PMID- 3037643 TI - [Computerized tomography and myelography findings following lumbar intervertebral disk surgery]. AB - 56 patients suspected of having suffered recurrent disc prolapse in the lumbar region were examined by CT and by myelography; the value of these examinations has been analysed. In 32 patients the findings were checked during surgery. There were difficulties in diagnosis, both for CT and myelography, where the findings were complex, i.e. in patients in whom there was postoperative fibrosis as well as disc prolapse and/or osseous spinal canal stenosis. As regards recurrence of prolapse, sensitivity for both methods was 83%. Postoperative scar formation can be clearly demonstrated by CT, but myelography produced a false positive diagnosis of recurrent prolapse in 17%. The changes were, in fact, caused by scar tissue formation. PMID- 3037644 TI - [MR tomography in intracranial meningiomas]. AB - The typical findings of intracranial meningiomas seen in 24 patients are described. The signal intensities at different acquisition parameters are analysed and optimal pulse sequences derived from these data. In 90% of the patients MR-tissue parameters were measured and correlated with the histological findings. Marked prolongation of T2-times was found in tumours showing regressive changes. PMID- 3037645 TI - [Iodine 131: biokinetics, radiation exposure and risk assessment with reference to the reactor accident at Chernobyl]. AB - Following the reactor accident at Chernobyl, this paper describes the biokinetics of radioiodine in man and discusses the radiation exposure resulting from intake of 131I. The risk of radiation-induced thyroid carcinomas and of congenital abnormalities is evaluated. Assuming a linear dose/risk relationship, one can calculate an increase in mortality from thyroid carcinomas amongst children in southern Germany of 100 to 101 per million children. For adults in southern Germany, and for the rest of the population in Germany, the figure is considerably lower. Gonadal dose from the 131I released is so small, compared with the annual natural radiation exposure, that it is not appropriate to discuss genetic effects. PMID- 3037646 TI - [Use of computers in radiology. Word processing, office technology and documentation--continuing the past or using newer technology?]. AB - The use of computers can result in drastic rationalisation in radiology with regard to text processing, office automation and documentation. The staff can be streamlined and work can be processed more rapidly, easier and more reliably, thus essentially improving effective productivity levels. Basing on the experience collected in handling the COMRAD system. Where the textrail is the most important software component, the article explains by means of the example of three different types of reports how the radiologist is assisted in his work by the computer. PMID- 3037647 TI - [Acute intramural small intestinal hematoma in hemophilia A]. PMID- 3037648 TI - Curative gelfoam embolisation of life-threatening bleeding from ascending colon diverticulum. A case report. PMID- 3037649 TI - [Fibrolamellar liver carcinoma. Diagnosis by imaging procedures]. PMID- 3037650 TI - Atypical calcified coarctation of the aorta. PMID- 3037651 TI - [Bilateral mandibular head metastases of a prostatic carcinoma]. PMID- 3037652 TI - [Local Thorotrast collection following paravascular injection of a contrast medium into the neck]. PMID- 3037653 TI - [TNM classification of malignant tumors: new edition 1987]. PMID- 3037654 TI - [Signet ring cell carcinoid of the appendix. Morphological, histochemical and immunohistochemical study of a case]. AB - A case of carcinoid tumor of the appendix with "signet ring" cells pattern and asymptomatic clinical course is presented. The tumor was detected during a Bilroth I gastrectomy procedure. Mucin-secreting cells with cytoplasms often containing argentaffin and argyrophil granules was the predominant histologic pattern. Lack of neuron-specific enolase activity and their positivity for lysozyme and EMA marker indicate an origin from crypt cell and not a neural crest source. After simple appendectomy, the patient was healthy 3 years following surgery. PMID- 3037655 TI - Angiotensin II: preferential efferent constriction? AB - In dogs with maximal subpression of endogenous angiotensin II (AII) production due to a high-salt diet and converting enzyme inhibition (CEI, SQ 14,225, 15 micrograms X kg-1 X min-1 i.v.), infusion of a subpressor dose of angiotensin II (1 ng X kg-1 X min-1) did not change contralateral kidney function. In the infused kidney, a decrease in renal blood (RBF) by 24% and glomerular filtration rate (GFR) by 9% with an increase in filtration fraction (FF) by 20% occurred. Similarly, the increase in single nephron (SN) RBF was greater than in SNGFR, thus rising SNFF by 8%. Glomerular capillary pressure (GCP) did not change significantly; a decrease by 20% in proximal tubule pressure thus resulted in an increase in delta HP by 22%. This increase counterbalanced the profound drop in ultrafiltration coefficient (Kf) (57%) making the decrease in GFR and SNGFR relatively small. Total arteriolar resistance (RT) rose by 26%, the rise being due mainly to an increase in efferent (RE, 50%) rather than afferent (RA, 4%) resistance. If the AII infusion was carried out during concomitant infusion of CEI and indomethacin (1 mg X kg-1 X min-1) or aspirin (5 mg X kg-1 X min-1), RBF decreased by 36%, GFR by 25%, thus increasing FF by 18%; corresponding SN values underwent similar changes. Drop in Kf amounted to 62% and hydraulic pressure difference (delta HP) increased by 11% with unchanged GCP. The increase in RT (72%) was now due to a very similar increase in both RA (68%) and RE (76%). In conclusion, a very small dose of AII exhibits-at least in superficial nephrons-a typical preferential efferent effect which disappears after inhibition of prostanoid synthesis, indicating a protective effect of vasodilatory prostaglandins mainly on the afferent arteriole. PMID- 3037656 TI - Effects of diethyldithiocarbamate and structural analogs in mice with systemic candidal infections. AB - Three different substituted dithiocarbamates: sodium diethyldithiocarbamate (DDTC), sodium dimethyldithiocarbamate (DmDTC), and sodium N-methyl-D-glucamine dithiocarbamate (NMG-DTC) were evaluated for their ability to combat the growth and development of three human pathogenic strains of Candida albicans, in vitro and in vivo, in mice. DDTC and DmDTC produced marked growth inhibition on agar plates, in vitro, of three different strains of Candida albicans, while NMG-DTC displayed little inhibitory effect. Low, nontoxic doses of each compound administered to immunosuppressed mice exhibited impressive therapeutic effects in treating candidiasis. NMG-DTC showed the best consistent therapeutic antifungal effect against Candida albicans in mice immunosuppressed with Solu-Medrol. Combinations of low doses of DDTC and Amphotericin-B appeared to be effective in treating systemic candidal infections, and the results suggested that these combinations may offer therapeutic advantages over those produced by the use of either agent alone. PMID- 3037657 TI - Some biological effects of calvatic acid and its analog on isolated hepatocytes. AB - Calvatic acid is a new antibiotic with both a carboxylic group and an azoxy function in the benzene ring. The relationship between structure and biological activity in this substance and one of its analog, phenylazoxyxyanide, was investigated. Isolated hepatocytes were used as an experimental system: the release of lactate dehydrogenase and the colchicine-binding activity of tubulin were first evaluated; effects were dose and time dependent. micromolar concentrations of these drugs determined a significant decrease of the glucose-6 phosphatase activity and a slight inhibition of aminopyrene demethylation. A toxic effect at the microsomal level and an interaction with the microtubular system are thus possible. PMID- 3037658 TI - A combined surgical approach to non-oat-cell pulmonary carcinoma with single cerebral metastasis. AB - Eighty consecutive patients with pulmonary non-oat-cell carcinoma and a single cerebral metastasis were followed for at least 5 years after therapy. Forty were treated by surgical excision at both sites of disease plus whole-brain irradiation in most cases (group 1). The remaining 40 patients, an observational cohort, were treated either by surgery at only one site of disease (usually craniotomy), whole-brain irradiation, chemotherapy, or some combination of these modalities (group 2). The 1-year survival in group 2 was 15%, and all were dead at 2 years. In group 1, hospital mortality was 1.5%, the 1-year survival rate 35%, the 2-year survival rate 25%, and the 5-year survival rate 12.5%. All the five year survivors were patients with N0 disease. In this subgroup of group 1, the five year survival was 20%. All patients surviving for more than 2 years were in group 1 and had a Karnofsky rating greater than 50 and N0 disease after staging. These data indicate that a combined surgical approach can be accomplished with low morbidity, low mortality, and increased survival rates, especially for patients with N0 disease who are vigorous enough to undergo the combined treatment. PMID- 3037659 TI - Fluid loading with whole blood or Ringer's lactate solution during CPR in dogs. AB - The influence of fluid loading during CPR on oxygen uptake and blood flow was investigated in 18 dogs (12-26 kg). Blood flows were measured with radioactive microspheres at 5 (control CPR), 13 and 20 min after the initiation of ventricular fibrillation and CPR. After 10 min, 9 dogs received a rapid infusion of whole blood (11 ml/kg, i.v.) and 9 dogs received Ringer's solution (11 ml/kg, i.v.). Oxygen uptake was not significantly altered by fluid loading with either whole blood or Ringer's solution. Fluid loading increased cardiac output 34% over the 5 min control value. However, left ventricular perfusion decreased to 74% and brain flow decreased to 65% of control. At 20 min, cardiac output and brain flow returned to near control values, while left ventricular flow remained low. Changes in organ perfusion can be explained in part by the concurrent changes in blood pressures. Central venous diastolic pressure increased significantly (from 9 to 14 mmHg) after fluid load. However, central arterial diastolic pressure did not rise proportionately (from 32 to 34 mmHg). Hence, the central A-V diastolic pressure difference decreased. Although fluid loading during CPR improved cardiac output, flow to the heart and brain decreased. Further, there was no increase in oxygen consumption, indicating that fluid loading did not improve metabolic status. PMID- 3037660 TI - The respiratory aspect of the treatment of brain injury associated with acute alcohol intoxication--results of an animal experiment. AB - The effects of spontaneous respiration and mechanical ventilation were examined by investigating the interaction between elevated intracranial pressure and alcohol intoxication. Ethanol (200 ml 48%) was infused in 11 young pigs with elevated cerebral pressure during mechanical ventilation (group 1), 7 young pigs with elevated cerebral pressure during spontaneous respiration (group 2), and 4 young pigs without elevated cerebral pressure during spontaneous respiration (group 3). While the behavior of intracranial pressure during mechanical ventilation in the animals from group 1 was inhomogeneous with a tendency to rise (29-34 mmHg), cerebral pressure (28-55 mmHg) increased drastically in the animals from group 2. This increase was associated with a sharp rise of Pa,CO2 (37.6-73.3 mmHg) and a decrease of Pa,O2 (74 mmHg to 13 mmHg). None of the animals in group 2 survived. Pa,CO2 also rose in alcoholized animals without elevated cerebral pressure (group 3) (41.9-63.9 mmHg); intracranial pressure, however, remained within the normal range. All animals in group 3 survived. Our findings indicate that elevated intracranial pressure and alcohol intoxication have a cumulative or potentiating effect on depression of the respiratory center. Respiratory depression can be prevented by mechanical ventilation and, therefore, a further rise of intracranial pressure generally avoided. PMID- 3037661 TI - Emergency endotracheal drug administration using aerosol. AB - Drug administration via the endotracheal route has previously involved direct installation in bolus form. However this necessitates an interruption in ventilation and, for the administration of therapeutic agents to the lungs, may result in failure of delivery to the appropriate sites. Aerosols have advantages over other routes of administration since the drug is delivered directly to its required site of action. Treatment with nebulised beta-2 agonists is the initial treatment of choice in acute severe asthma and a small number of such patients require immediate intubation and ventilation. We describe a technique for aerosol delivery using a nebuliser which can be used in intubated patients during manual intermittent positive pressure ventilation using a bag-valve resuscitator. PMID- 3037662 TI - Differential lung ventilation: a review and 2 case reports. AB - The respiratory parameters of some of the patients with acute respiratory failure deteriorates while using conventional ventilation. These patients suffer unilateral lung disease and the failure to respond favourably to therapy is due to increased intrapulmonary shunt. There is a reflex vasodilation in the injured lung. Functional residual capacity is reduced in the injured lung and the compliance decreases. Gas flow is then deviated to the other lung, thus increases alveolar collapse and decreases regional compliance in the injured lung. These events cause severe hypoxemia. We present here two cases with unilateral lung disease that failed to respond to conventional mechanical ventilation. Asynchronized differential lung ventilation was found to be the therapeutic answer to the problem. We discuss the pathophysiology of unilateral lung injury and the physiology of differential lung ventilation. PMID- 3037663 TI - Effect of alterations in frequency, inspiratory time, and airway pressure on gas exchange during high frequency jet ventilation in dogs with normal lungs. AB - High frequency jet ventilation (HFJV) is becoming increasingly useful for providing respiratory support in patients with normal lungs during operative procedures, and also has been advocated as a technique for ventilating patients during cardiopulmonary resuscitation. We studied the effect of frequency, percent inspiratory time (I/E ratio), peak airway pressure, and airway pressure difference (peak-PEEP) during HFJV as operational variables on the efficacy of gas exchange in dogs with normal lungs. We observed that at a constant peak airway pressure and percent inspiratory time, PaCO2 generally increases as frequency rises above 100/min. In contrast, PaCO2 generally decreases as percent inspiratory time is reduced at a constant frequency and peak airway pressure. In addition, increasing peak airway pressure and airway pressure difference are associated with lower levels of PaCO2. Arterial oxygenation was adversely affected by frequencies above 300/min, but was otherwise not influenced by alterations in frequency, percent inspiratory time, or airway pressure. PMID- 3037664 TI - Theoretical effects of fluid infusions during cardiopulmonary resuscitation as demonstrated in a computer model of the circulation. AB - Recent studies have shown the potential adverse effects of venous volume loading on blood flow during closed chest cardiopulmonary resuscitation (CPR). To examine the effect of arterial and venous infusions, we employed a published computer simulation of the circulation during CPR. This model uses computer simulated electrical networks to model the heart and great vessels. CPR was modeled with compressions at a rate of 80/min and a force of 80 mmHg. Fluid infusions, simulated as current pulses into the abdominal aorta and superior vena cava, were given to measure their effect on myocardial and cranial blood flow. With 600 ml/min infusions into the abdominal aorta, there was a 12% peak increase in myocardial flow and a 3.8% peak increase in cranial flow. Every 100 ml/min increase in infusion from 0 to 900 ml/min produced a 1.4 ml/min linear increase in myocardial flow and a 4.2 ml/min linear increase in cranial flow. In agreement with previous CPR model studies, simulated vasoconstriction of abdominal and lower extremity vessels resulted in increased myocardial and cranial flows. As resistance of these vessels was increased, abdominal aortic infusions resulted in greater flow augmentations. In contrast to arterial results, infusions at 600 ml/min into the vena cava resulted in a 2.2% decrease in myocardial flow and a 0.62% decrease in cranial flow. Rise and fall times for initiation and cessation of flow augmentations were equal to four compression cycles. We conclude that these findings demonstrate the theoretical benefits of rapid arterial infusions during CPR with increases in myocardial and cranial blood flow. This method may provide an early temporary adjunct to myocardial perfusion during CPR. PMID- 3037665 TI - Sympathetic innervation of human tracheal and bronchial smooth muscle. AB - We have assessed functional and structural evidence for sympathetic innervation of human tracheal and bronchial smooth muscles. In the bronchus, functional evidence for such innervation was shown by the following results: a leftward shift in the noradrenaline inhibitory dose-response curve by the neuronal uptake blocker imipramine; the inhibition of uptake of 3H-L-noradrenaline by the neuronal uptake blockers imipramine, cocaine and phenoxybenzamine; and the induced release of noradrenaline and its metabolites by tyramine and electrical field stimulation using nerve-specific parameters; and selective inhibition of field-stimulated contractions by isoprenaline. Structural evidence was shown by the presence of small dense-cored vesicles containing nerve varicosities, occasionally in close proximity to morphologically characteristic cholinergic nerve-endings. This suggests the possibility of presynaptic modulation of acetylcholine release by noradrenaline. PMID- 3037666 TI - Human papillomavirus and cervical intraepithelial neoplasia as sexually transmitted diseases. PMID- 3037667 TI - [A 64-year-old woman with hemolytic anemia, thrombocytopenia and neurologic disorders]. PMID- 3037668 TI - Use of vitamin D analogs in renal osteodystrophy. PMID- 3037669 TI - [Recurrent oral ulceration: is it of viral etiology?]. PMID- 3037670 TI - [Primary carcinoma of the liver: study of 85 cases of necropsy]. PMID- 3037671 TI - [Glial tissue and pathology of the nervous system]. PMID- 3037672 TI - [Role of the involvement of the reticular formation in the dementia of Parkinson's disease]. AB - A post mortem study of 14 cases of Parkinson's disease was reported. Seven patients were mentally deteriorated and 7 were not, retrospectively. The semiquantitative study of the cortical lesions of senile dementia of Alzheimer type did not show any recognizable difference between the 2 groups. A greater degree of neuronal cell loss in the nucleus basalis of Meynert in the mentally deteriorated group was confirmed. The incidence of Lewy bodies in the brain stem reticular formation was remarkably increased in the mentally deteriorated group. A study of the reticular formation, including the raphe nuclei and the magnocellular nuclei of mid pons and upper medulla, revealed a significant higher incidence of Lewy bodies throughout the whole reticular formation in the mentally deteriorated group. The possible role of the lesions of the ascending reticular activating system in the mental deterioration is discussed. PMID- 3037673 TI - [Cholinergic chemical transmission. Mechanisms of control]. AB - The isolation of fractions of purified synaptic constituents has permitted to localize the enzymes involved in cholinergic mechanisms and to evaluate the acetylcholine concentration of vesicular stores and of the cytoplasmic pool where transmitter is synthetized. The isolation of cholinergic synaptosomes allowed study the mechanisms and regulations controlling the release process. The depletion of cytoplasmic acetylcholine elicited by stimulation is discussed in parallel to the role of synaptic vesicles. Presynaptic membrane proteins seem to be directly involved in the release of the transmitter. PMID- 3037674 TI - Animal models for acquired immunodeficiency syndrome. AB - Substantial advances have already been made in the understanding of acquired immunodeficiency syndrome (AIDS). The major issues for AIDS research during the next few years must be practical ones: the development of a safe, effective vaccine for individuals not yet infected with the causative virus and the development of drug therapies for those already infected. Suitable animal models will be needed for studies designed to achieve these goals. Areas of investigation in animal models can be divided into four categories on the basis of increasing direct relevance to AIDS in humans: retroviruses that have no obvious, close relation to human immunodeficiency virus (HIV) but can induce chronic diseases with manifestations that include immunologic abnormalities; ungulate lentiviruses; HIV-related viruses of Old World primates; and HIV infection of chimpanzees. It is hoped that important research developments in experimental models can be quickly extrapolated to human AIDS. PMID- 3037675 TI - Epidemiology of rotaviral infection in adults. AB - Although classic rotaviral gastroenteritis occurs in children between the ages of six and 24 months, infection with rotavirus is common in all age groups, including adults. Virtually all adults have been infected, as is demonstrated by the presence of serum antibodies, but previous infection does not protect against new infection with the same or a different serotype. Rotaviral infection of adults is seen in five settings: secondary contacts from pediatric cases, with variable attack rates in adults; waterborne outbreaks, which are often characterized by higher attack rates in adults than in children; travelers' diarrhea; epidemic spread in isolated or closed populations, often in the absence of contact with children; and endemic infections, which may account for 5%-10% of sporadic cases of diarrhea in adults. Frequent asymptomatic infections with rotavirus occur, and they may be important in the epidemiology of the disease. Although rotaviral infections in adults tend to be milder than those in children, death due to rotaviral infection in adults have been reported. PMID- 3037676 TI - Rifabutin (ansamycin LM 427): a new rifamycin-S derivative for the treatment of mycobacterial diseases. AB - Rifabutin (ansamycin LM 427), a semisynthetic spiropiperidyl derivative of rifamycin S, shows good in vitro activity against most mycobacterial species, including Mycobacterium avium complex. In animal models, the drug is more active against both Mycobacterium tuberculosis and Mycobacterium leprae than in rifampin, and studies indicate that rifabutin is active against some rifampin resistant strains of both species. The drug has a long half-life (16 hr) in humans and a marked tissue tropism, with tissue levels five- to 10-fold higher than that in the serum. In animals rifabutin is no more toxic than rifampin. A large experience from the compassionate use of rifabutin for life-threatening mycobacterial infections in humans, most commonly disseminated M. avium complex disease in patients with AIDS, has also indicated relative drug safety. Although some data suggest that rifabutin is effective, firm conclusions about drug efficacy await results from controlled clinical trials. PMID- 3037677 TI - The moveable genome. Weismann's doctrine and new models for speciation. PMID- 3037678 TI - The null hypothesis in vertebrate evolution. PMID- 3037679 TI - The 'bio-bang'. PMID- 3037680 TI - Antibodies to the herpes simplex virus type-2-induced tumor-associated antigen AG 4 as markers of recurrence in cervical cancer. AB - This study was designed to evaluate the potential clinical value of AG-4 antibodies in patients with established cervical cancer, in terms of the assessment of prognosis before treatment, and monitoring the course of the cancer process during long-term follow-up. Forty-seven patients were investigated during a median follow-up time of 5.4 years (range 3.5-6.2). During this time, 25 developed recurrent disease. Serum was obtained from all patients before treatment and at regular intervals during follow-up and assayed for antibodies to AG-4. The presence of AG-4 antibodies during follow-up was associated with recurrent disease (p less than 0.001). In most cases the presence of AG-4 antibodies preceded the clinical diagnosis of recurrence, with particularly long lead times in the late recurrences. Serial determinations of AG-4 antibodies in patients with invasive cervical cancer may be of clinical value for monitoring the course of the disease during follow-up, thus allowing for earlier application of treatment. PMID- 3037681 TI - Novel cholinesterase expression in the HuH-7 cell line. AB - On polyacrylamide gradient gel electrophoresis, normal serum cholinesterase was separated into seven isozymes (I-VII from the anodic to the cathodic side). The enzyme of the conditioned medium of the HuH-7 cell line, established from a human hepatoma, had two main isozymes. The one migrating faster was located slightly to the cathodic side of band II of the normal serum enzyme, and the other, a slower one, electrophoresed at the same position as that of band VI of the normal serum enzyme. Aside from these two isozymes, a faint band with enzyme activity sometimes appeared at a position just cathodic or very close to the position of band I of the normal serum isozymes. The effect of some inhibitors and activators on both the normal serum enzyme and the enzyme of the conditioned medium was similar, but lectin-binding properties of the two enzymes were different with Ricinus communis agglutinin I, concanavalin A and wheat germ agglutinin. These results suggest that the difference in sugar moieties of both enzymes is expressed in D-galactose, D-mannose and N-acetylglucosamine. PMID- 3037682 TI - Automated method for the determination of angiotensin-converting enzyme in serum. AB - A method for the determination of angiotensin-converting enzyme in serum (S-ACE; EC 3.4.15.1.) with use of 3-(2-furylacryloyl)-L-phenylalanyl-glycyl-glycine (FAPGG) as substrate has been adapted for the Cobas Bio microcentrifugal analyser. The method allows 24 determinations per hour in a sample volume of 28 microliters with a within run precision of less than 3% and a between run precision of less than 5%. The reaction is linear up to at least 470 U/l. The reference interval in 92 blood donors has been determined to 14-110 U/l. The method correlates well with the manual method of Hurst & Lovell-Smith (r = 0.982). We have found the method excellently suited for routine assay. PMID- 3037683 TI - The influence of ispaghula husk and lactulose on the in vivo and the in vitro production capacity of short-chain fatty acids in humans. AB - To evaluate factors influencing the short-chain fatty acid (SCFA) concentrations in stools, three different experiments were performed: faecal concentrations of SCFA at defecation were determined by gas liquid chromatography in nine healthy volunteers on a free diet. SCFAs were 114 +/- 15.0 mmol/l (means +/- SD). The coefficient of variation (CV) of the assay was 4-15%, the intraindividual CV 12 33%, and the interindividual CV 11-29%. On incubation of faeces at 37 degrees C concentrations of SCFA doubled in 6 h and rose fourfold in 72 h. In three volunteers the experiments were extended by adding ispaghula husk or lactulose to the diet for two 14-day periods each; no change in faecal SCFA concentrations was seen, either at defecation or after incubation. When ispaghula husk or lactulose was added to faeces in an in vitro incubation system, the concentrations of SCFA were five times higher than those of controls. We conclude that instant handling of faeces is essential for determinations of SCFA concentrations to obtain interpretable and comparable results; that determination of total SCFA output is of limited value; that addition of fibre to the diet does not influence faecal SCFA concentrations; and that the capacity for SCFA production in faeces is large provided a sufficient amount of substrate is available. PMID- 3037684 TI - Expression of CR1 and CR2 complement receptors following Epstein-Barr virus infection of Burkitt's lymphoma cell lines. AB - Epstein-Barr virus (EBV) infection of human B lymphocytes involves a specific receptor closely associated with, or identical to, the C3d complement receptor, CR2. Thus, 25 out of 29 EBV-positive Burkitt's lymphoma (BL) cell lines but none of 15 EBV-negative BL lines were found to express C3 receptors. Furthermore, in vitro infection with EBV of six EBV-negative cell lines resulted in the expression of C3 receptors in association with that of EBV-determined nuclear antigen (EBNA). Rosette assays using erythrocytes coated with human C3b, C3bi, and C3d, inhibition of rosette formation with anti-receptor antibodies, and flow cytometry analysis of stained cells demonstrated that EBV-converted lines expressed C3b and C3d receptors, CR1 and CR2. Anti-receptor antibodies recognized an average of 40,700 anti-CR1 and 140,000 anti-CR2 binding sites on an EBV converted line (BL41/B95), whereas no specific binding occurred on the corresponding EBV-negative (BL41) cells. Because CR1 and CR2 are involved in B cell proliferation and/or differentiation, enhanced expression of C3 receptors following the interaction between EBV and B cells and/or subsequent infection of the cells by EBV may provide a basis for positive control of B lymphocyte proliferation by EBV. PMID- 3037685 TI - Suppression of superoxide generation by normal polymorphonuclear leukocytes preincubated in plasma from patients with Felty's syndrome. AB - Polymorphonuclear leukocytes (PMN) isolated from patients with Felty's syndrome (FS) generate fewer superoxide anions (O-2) upon stimulation with fmet-leu-phe than PMN from normal controls or patients with rheumatoid arthritis (RA). In this study, plasma samples were obtained from 12 patients with RA and 12 patients with FS. Incubation of normal PMN in plasma from Felty patients resulted in a significant reduction in both the rate and total quantity of O-2 generation when activated with fmet-leu-phe. This was not observed with plasma from RA patients. The capacity of a plasma sample to suppress O-2 generation correlated with plasma IgG-PMN-binding activity (IgG-PBA) and, to a lesser extent, with the content of circulating immune complexes (CIC). These data suggest that IgG-PBA and possibly CIC have a pathogenetic role in both qualitative and quantitative defects in PMN in Felty patients. PMID- 3037686 TI - Activation of latent collagenase purified from human leukocytes. AB - Latent human leukocyte collagenase was isolated to apparent homogeneity by a simple and rapid method. Isolation was accomplished by gel filtration on Sepharose 6B and ion exchange chromatography on QAE Sephadex A-50 followed by affinity chromatography on Cibacron Blue Sepharose. The purified latent enzyme exhibits an apparent molecular weight of 70 kD as estimated by SDS-polyacrylamide gel electrophoresis. Reduction with dithiothreitol does not change the mobility of the latent human leukocyte collagenase on SDS-polyacrylamide gel electrophoresis, indicating that the enzyme consists of a single polypeptide chain. The enzyme could be activated by trypsin and thiol reagents such as phenylmercuric chloride and N-ethylmaleimide. Upon activation by trypsin a 54 kD polypeptide was formed from the 70 kD latent enzyme. Concomitant with the activation by thiol reagents, no loss of molecular weight was detected. Inactivated trypsin, i.e. phenylmethyl sulfonyl-trypsin or soybean trypsin inhibitor treated trypsin, was not able to activate latent human leukocyte collagenase. The results support the concept that latent human leukocyte collagenase exists as a proenzyme and thiol-dependent activation occurs through conformational perturbation in the proenzyme molecule. PMID- 3037687 TI - No association between antibodies to HTLV-1 and polymyositis, rheumatoid arthritis and SLE. PMID- 3037688 TI - [Parvovirus infections detected in swine in the years 1985/86]. PMID- 3037689 TI - [Regulation of cell surface receptors]. PMID- 3037690 TI - The molecules of visual excitation. PMID- 3037691 TI - Survival after ovarian granulosa and theca cell tumours. AB - Ninety-two cases of ovarian granulosa and theca cell tumours were recorded in the Tayside Gynaecological Tumour Registry during the 30 year period 1948-78. A high follow-up rate was achieved through this centralised registry and exhaustive annual recall. At 20 years after initial treatment patient actuarial survival was 92% for the theca cell tumours and 34% for granulosa cell tumours. PMID- 3037692 TI - ACTH deficiency: hypothalamic or pituitary in origin? AB - The level within the hypothalamic-pituitary axis at which isolated ACTH deficiency occurs, can be more clearly identified by assessing this axis with corticotrophin-releasing factor (CRF) and insulin induced hypoglycaemia. We report a case where lack of response to both tests, suggests a pituitary origin for this rare endocrine deficiency. PMID- 3037693 TI - A promoter with an internal regulatory domain is part of the origin of replication in BPV-1. AB - Extrachromosomal elements that are stably maintained at a constant copy number through cell doublings are a good model system for the study of the regulation of DNA replication in higher eukaryotes. Previous studies have defined both cis and trans functions required for the regulated plasmid replication of the bovine papilloma virus in stably transformed cells. Here, a sequence known to be a cis dominant element of the replication origin of the plasmid is shown to contain a promoter for transcription. Both in vitro and in vivo assays have been used to define this promoter and show that a sequence located just 3' to the transcriptional start site is required for activity. This DNA sequence element, which has been defined through deletions, coincides with a binding site for a cellular factor and is also required for a functional origin of replication. Possible models for how a transcription factor may play a role in the regulation of DNA replication are discussed. PMID- 3037694 TI - Retroviruses and mouse embryos: a rapid model for neurovirulence and transplacental antiviral therapy. AB - A murine model in which neurotropic retroviral infection can be studied over short periods of time was developed. Microinjection of Cas-Br-E virus into midgestation mouse embryos caused paralysis and death within 25 days after birth, in contrast to virus-infected neonates which develop disease only after 4 months. To evaluate whether antiviral drugs could cross the placental barrier and influence the course of the disease, the drug 3'-azido-3'-deoxythymidine (AZT) was administered to infected embryos through the drinking water of pregnant females. AZT treatment markedly retarded the onset and course of virus-induced central nervous system disease, permitting animals to survive beyond 4 months of age. These results are evidence for effective antiviral treatment during gestation and in the perinatal period and are of potential significance for the management of maternal transmission of the acquired immune deficiency syndrome (AIDS) virus. PMID- 3037695 TI - Divalent cations directly affect the conductance of excised patches of rod photoreceptor membrane. AB - Phototransduction in rod cells is likely to involve an intracellular messenger system that links the absorption of light by rhodopsin to a change in membrane conductance. The direct effect of guanosine 3',5'-monophosphate (cGMP) on excised patches of rod outer segment membrane strongly supports a role for cGMP as an intracellular messenger in phototransduction. It is reported here that magnesium and calcium directly affect the conductance of excised patches of rod membrane in the absence of cGMP and that magnesium, applied to intact rod cells, blocks a component of the cellular light response. The divalent cation-suppressed conductance in excised patches showed outward rectification and cation-selective permeability resembling those of the light-suppressed conductance measured from the intact rod cell. The divalent cation-suppressed conductance was partly blocked by a concentration of the pharmacological agent L-cis-diltiazem that blocked all of the cGMP-activated conductance. Divalent cations may act, together with cGMP, as an intracellular messenger system that mediates the light response of the rod photoreceptor cell. PMID- 3037696 TI - A physiological basis for a theory of synapse modification. AB - The functional organization of the cerebral cortex is modified dramatically by sensory experience during early postnatal life. The basis for these modifications is a type of synaptic plasticity that may also contribute to some forms of adult learning. The question of how synapses modify according to experience has been approached by determining theoretically what is required of a modification mechanism to account for the available experimental data in the developing visual cortex. The resulting theory states precisely how certain variables might influence synaptic modifications. This insight has led to the development of a biologically plausible molecular model for synapse modification in the cerebral cortex. PMID- 3037697 TI - Selective disruption of gap junctional communication interferes with a patterning process in hydra. AB - The cells that make up the body column of hydra are extensively joined by gap junctions, capable of mediating the rapid exchange of small hydrophilic molecules between the cytoplasms of neighboring cells. Both the rate of transfer of small molecules through the gap junctions and the rate of return of gap junction coupling after grafting experiments are sufficiently rapid to mediate events in the patterning of hydra tissue. Antibodies to the major rat liver gap junction protein (27,000 daltons) recognize a gap junction antigen in hydra and are effective in eliminating junctional communication between hydra cells. The antibodies perturb the head inhibition gradient in grafting operations, suggesting that cell-cell communication via gap junctions is important in this defined tissue patterning process. PMID- 3037698 TI - T-cell tumor elimination as a result of T-cell receptor-mediated activation. AB - It has recently been shown that activation of murine T-cell hybridomas with antigen inhibits their growth in vitro. The "suicide" of these neoplastic T cells upon stimulation with antigen suggested the possibility that activation via the antigen-specific receptor could also inhibit the growth of neoplastic T cells in vivo. To test this, mice were subcutaneously inoculated with antigen-specific T cell hybridomas and then treated intraperitoneally with antigen. Administration of the appropriate antigen immediately after inoculation with the T-cell hybridoma abrogated tumor formation; antigen administered after tumors had become established decreased the tumor burden and, in a substantial fraction of animals, led to long-term survival. The efficacy of antigen therapy was due to both a direct inhibitory effect on tumor growth and the induction of host immunity. These studies demonstrate the utility of cellular activation as a means of inhibiting neoplastic T-cell growth in vivo and provide a rationale for studying the use of less selective reagents that can mimic the activating properties of antigen, such as monoclonal antibodies, in the treatment of T-cell neoplasms of unknown antigen specificity. PMID- 3037699 TI - Debate over potential AIDS drug. PMID- 3037700 TI - Cellular mechanisms of epilepsy: a status report. AB - The cellular phenomena underlying focal epilepsy are currently understood in the context of contemporary concepts of cellular and synaptic function. Interictal discharges appear to be due to a combination of synaptic events and intrinsic currents, the exact proportion of which in any given neuron may vary according to the anatomic and functional substrate involved in the epileptic discharge and the epileptogenic agent used in a given model. The transition to seizure appears to be due to simultaneous increments in excitatory influences and decrements in inhibitory processes--both related to frequency-dependent neuronal events. A variety of specific hypotheses have been proposed to account for the increased excitability that occurs during epileptiform activity. Although each of the proposed mechanisms is likely to contribute significantly to the epileptic process, no single hypothesis provides an exclusive unifying framework within which all kinds of focal epilepsy can be understood. The spread of epileptic activity throughout the brain, the development of primary generalized epilepsy, the existence of "gating" mechanisms in specific anatomic locations, and the extrapolation of hypotheses derived from simple models of focal epilepsy to explain more complex forms of human epilepsy, all are not yet fully understood. PMID- 3037701 TI - Variable occurrence of the nrdB intron in the T-even phages suggests intron mobility. AB - The bacteriophage T4 nrdB gene, encoding nucleoside diphosphate reductase subunit B, contains a self-splicing group I intervening sequence. The nrdB intron was shown to be absent from the genomes of the closely related T-even phages T2 and T6. Evidence for variable intron distribution was provided by autocatalytic 32P guanosine 5'-triphosphate labeling of T-even RNAs, DNA and RNA hybridization analyses, and DNA sequencing studies. The results indicate the nonessential nature of the intron in nrdB expression and phage viability. Furthermore, they suggest that either precise intron loss from T2 and T6 or lateral intron acquisition by T4 occurred since the evolution of these phages from a common ancestor. Intron movement in the course of T-even phage divergence raises provocative questions about the origin of these self-splicing elements in prokaryotes. PMID- 3037702 TI - Mapping patterns of c-fos expression in the central nervous system after seizure. AB - A dramatic and specific induction of c-fos was observed in identifiable neuronal populations in vivo after administration of the convulsant Metrazole. This effect was time- and dose-dependent and was abolished by prior treatment with the anticonvulsant drugs diazepam or pentobarbital. About 60 minutes after administration of Metrazole, c-fos messenger RNA reached a maximum and declined to basal levels after 180 minutes. A further decrease below that in normal brain was observed before a return to basal levels after 16 hours. While Metrazole still elicited seizures during this period, reinduction of c-fos was largely refractory. At 90 minutes, c-fos protein was observed in the nuclei of neurons in the dentate gyrus, and in the pyriform and cingulate cortices. Subsequently, c fos protein appeared throughout the cortex, hippocampus, and limbic system. Thus, seizure activity results in increased c-fos gene expression in particular subsets of neurons. PMID- 3037703 TI - Cloning of human mineralocorticoid receptor complementary DNA: structural and functional kinship with the glucocorticoid receptor. AB - Low-stringency hybridization with human glucocorticoid receptor (hGR) complementary DNA was used to isolate a new gene encoding a predicted 107 kilodalton polypeptide. Expression studies demonstrate its ability to bind aldosterone with high affinity and to activate gene transcription in response to aldosterone, thus establishing its identity as the human mineralocorticoid receptor (hMR). This molecule also shows high affinity for glucocorticoids and stimulates a glucocorticoid-responsive promoter. Together the hMR and hGR provide unexpected functional diversity in which hormone-binding properties, target gene interactions, and patterns of tissue-specific expression may be used in a combinatorial fashion to achieve complex physiologic control. PMID- 3037704 TI - Functional analysis of a complementary DNA for the 50-kilodalton subunit of calmodulin kinase II. AB - The calcium-calmodulin-dependent protein kinase II is a major component of brain synaptic junctions and has been proposed to play a variety of important roles in brain function. A complementary DNA representing a portion of the smaller 50 kilodalton subunit of the rat brain enzyme has been cloned and sequenced. The calmodulin-binding region has been identified and a synthetic analog prepared that binds calmodulin with high affinity in the presence of calcium. Like the 50 kilodalton kinase polypeptide, the concentration of the messenger RNA varies both neuroanatomically and during postnatal development of the brain. The broad tissue and species cross-reactivity of the complementary DNA suggests that the 50 kilodalton subunit found in rat brain is evolutionarily conserved and is the product of a single gene. PMID- 3037705 TI - Identification of a family of muscarinic acetylcholine receptor genes. AB - Complementary DNAs for three different muscarinic acetylcholine receptors were isolated from a rat cerebral cortex library, and the cloned receptors were expressed in mammalian cells. Analysis of human and rat genomic clones indicates that there are at least four functional muscarinic receptor genes and that these genes lack introns in the coding sequence. This gene family provides a new basis for evaluating the diversity of muscarinic mechanisms in the nervous system. PMID- 3037706 TI - Opportunistic infections in the acquired immune deficiency syndrome. AB - The acquired immune deficiency syndrome (AIDS) results in a T lymphocyte deficiency and, consequently, a susceptibility to opportunistic infections with organisms that previously were better known as causes of disease in patients with Hodgkin's disease or recipients of immunosuppressive therapy. Pneumocystis carinii, Candida albicans, and cytomegalovirus (CMV) are among the most frequently observed pathogens in AIDS patients, followed by atypical mycobacteria, Toxoplasma gondii, Mycobacterium tuberculosis, Salmonella sp, and herpes simplex virus. Cryptosporidia sp, rarely encountered in iatrogenically immunosuppressed patients, have also emerged as important pathogens in the setting of AIDS. Although some opportunistic infections in AIDS patients can be effectively treated, others are only temporarily suppressed or do not respond to the drugs currently available. The severity of the underlying immune deficit remains the most important prognostic factor, and opportunistic infection remains the cause of death in most AIDS patients. PMID- 3037707 TI - Granular cell tumor of the esophagus: a study of seven cases diagnosed by histologic examination of endoscopic biopsies. AB - Among the benign tumors of the esophagus, the discovery of a granular cell tumor is exceptional. Although endoscopy permits recognition of this tumor and supports its benign nature, only histologic examination of the endoscopic biopsy specimen permits one to determine its precise nature. The endoscopic appearance of esophageal granular cell tumor ranges from a plaque-like thickening of the mucosa to a polypoid mass. We describe here seven cases of granular cell tumor of the esophagus diagnosed by histologic examination of endoscopic biopsy specimens. We believe the actual incidence of this tumor in this location might be greater if every esophagus were given as much attention at endoscopic examination and at autopsy. PMID- 3037708 TI - [Surgical treatment of lung cancer in older age groups]. PMID- 3037709 TI - Anti-varicella zoster immunoglobulin and herpes simplex infections. PMID- 3037710 TI - Viruses excreted in neonatal stools. AB - Faecal specimens from 122 infants in the Neonatal Unit at Ga-Rankuwa Hospital were examined by electron microscopy for the presence of virus. In total 40% of the neonates were excreting virus. Rotavirus was the commonest, found in 33.5% of the infants, with small round viruses (SRVs) found in 12.3% and adenovirus seen in 1 baby. RNA analysis of the rotavirus genome revealed a similar electrophoretype in all specimens. Most of the babies infected with rotavirus were less than 7 days old, and very little excretion occurred after 21 days of age. Only 8% of the babies excreting virus had any clinical signs of infection. Rotavirus infection appears to be endemic and SRV infection common in the Neonatal Unit at Ga-Rankuwa Hospital. PMID- 3037711 TI - Transhepatic portal sampling for preoperative localization of insulinomas. AB - The data of 72 patients with pancreatic insulinomas were analyzed. Twenty-one were obtained from personal experience and 51 from a review of the literature. In all instances, detailed information about insulin levels in the portal tree as obtained by means of a transhepatic portal sampling (THPS) and localization of the tumor as found during the surgical procedure was available. Five different criteria were compared for defining the peak insulin concentrations regarded as indicative of tumor localization: 1, peak above 500 milliunits per liter; 2, peak above 200 milliunits per liter; 3, peak 2.5 times higher than the lowest portal value; 4, peak 2.3 time higher than the distal mesenteric value, and 5, peak higher than mean portal concentration plus or minus 2 standard deviation. Criterion 5, associated with sample numbers larger than 15 and catheterization of the cephalic veins, provided the best results for obtaining valuable information from THPS. PMID- 3037712 TI - Ulnar neuropathy in a patient with multiple schwannomas of the ulnar nerve. PMID- 3037713 TI - [The prematurely born infants can be a very great responsibility]. PMID- 3037714 TI - [Activity of cell enzymes during treatment of bronchial asthma patients with euphylline and hydrocortisone]. AB - Two groups of patients with bronchial asthma (of average severity and a severe course) and a group of healthy persons were examined. Changes in dehydrogenase activity in the time course of asthma development associated with the depletion of the adaptation mechanisms, and the compensation of these disorders as a result of glucocorticosteroid therapy were established. AP activity in neutrophils could serve as an informative sign indicative of asthma progression and showing the efficacy of hormonotherapy. Various shifts in the metabolic activity of enzymes ensuring the efficacy of phagocytosis with relation to a degree of asthmatic symptoms and therapeutic modalities were revealed. PMID- 3037715 TI - Deleterious effects of adrenocorticotrophic hormone administration during late pregnancy upon offspring somatic, neurological, and sexual development in mice. AB - The influence of administration of adrenocorticotrophic hormone (ACTH) during days 12-17 of pregnancy upon somatic, neurological, and neuromuscular development of offspring in mice was studied. The effects upon the onset of puberty in female offspring was also examined. Litters from mice given the higher of two doses of ACTH (1 IU/day or 8 IU/day) showed lower body weights at birth and weaning than controls. This treatment also increased pre- and postnatal mortality rates, although not significantly. Litters from mice treated with either dose of ACTH showed retarded development of the forelimb and hindlimb grasp reflexes, the body righting reflex, the auditory startle response, and eye opening. Although ear opening was delayed in litters from ACTH-treated mice, results did not achieve statistical significance. Study of female offspring housed in small groups revealed that one indicator of puberty, vaginal opening, was delayed in female offspring of ACTH-treated mice. Experiments were conducted to identify factors mediating this syndrome: ACTH did not depress maternal food intake or alter the length of pregnancy, therefore fetal undernutrition or premature birth can be excluded as mediating factors. All litters were fostered to untreated mice to control for postnatal factors influencing development. As ACTH cannot cross the placenta, the syndrome is likely to result from in utero exposure to abnormally high concentrations of glucocorticosteroids of maternal origin. It is concluded that such alterations to the fetal environment can exert a deleterious influence upon somatic, neurological, and sexual development, and that hormones of the maternal pituitary-adrenocortical axis may naturally act to regulate general development of the fetus. PMID- 3037716 TI - Enhanced susceptibility of mouse embryos heterozygous for oligosyndactyly (Os/+) to mitomycin C-induced skeletal abnormalities. AB - The mouse mutation, oligosyndactyly (Os), results in syndactyly, muscle anomalies, and deficiency of nephrons in heterozygous animals and early embryonic lethality in homozygotes. Since the homozygous lethality results from mitotic arrest with intact spindles at the time of implantation, we have hypothesized that the heterozygous manifestations may result from impairment of cell proliferation in regions with high proliferative rates. To test this hypothesis, Os/+ and +/+ mouse embryos at 6.5 days of gestation were exposed to mitomycin C (MMC), an agent that causes a high degree of embryonic cell death which is "compensated" for by a period of rapid cell proliferation. 17.9% of MMC-treated +/+ fetuses had fused vertebrae, a significant increase over untreated fetuses, and this frequency was further increased to 33.6% in MMC-treated Os/+ fetuses. Saline treated Os/+ and +/+ fetuses showed the same low rate (0-3%) of vertebral fusion. These results indicate that Os/+ embryos have an increased sensitivity to the vertebral fusion-inducing effect of MMC at 6.5 days of gestation, a finding compatible with the hypothesis that rapid cell proliferation may be impaired in Os/+ embryos and fetuses. PMID- 3037717 TI - Dysfunctional activated protein C (PC Cadiz) in a patient with thrombotic disease. AB - The partial characterization of a dysfunctional protein C (PC), provisionally named "PC Cadiz", in a 45-year-old male patient suffering from recurrent venous thrombosis is described. The only defect found in laboratory assays for haemostasis and hepatic function was a half normal level of both amidolytic and anticoagulant protein C activity, measured by different functional assays that use thrombin-thrombomodulin complex and a snake venom to activate protein C. Protein C antigen was always found to be within normal levels. Two young daughters of the propositus were found to have the same defect. Double-crossed immunoelectrophoresis, performed in the presence and absence of Ca2+ in the first dimension, showed no clear differences between patient and control PC. PC adsorption to barium salts was also found to be normal. Measurement of the PC activation peptide in the barium citrate eluates after PC activation showed no significant differences between patient and 10 normal controls, the concentration of this peptide being very similar to that of PC zymogen in the same eluates before PC activation. These results indicate that this abnormal PC is able to be normally activated by thrombin-thrombomodulin complex but does not exhibit serine protease activity, probably due to a defect in the PC molecule near the active site center. PMID- 3037718 TI - In vitro and in vivo pharmacological profiles of the PAF receptor antagonist SRI 63-675. AB - We have examined a recently developed PAF antagonist SRI 63-675 (dimethyl tetrahydrofuran-methoxyphosphinyloxy-ethylquinolinium ) for its ability to inhibit several major PAF-induced physiological responses. The compound was a potent inhibitor of PAF-induced platelet aggregation in platelet rich plasma obtained from humans, guinea pigs, and rabbits, with IC50 values of 3.43, 0.25, and 0.97 microM, respectively. SRI 63-675 did not inhibit ADP, collagen nor epinephrine-induced human platelet aggregation. The IC50 for inhibition of PAF receptor binding to human platelets was 0.37 microM. In the rat SRI 63-675 inhibited 0.1 microgram kg-1 i.v. PAF-induced hypotension, with an ED50 of 32 micrograms kg-1 i.v. Using the same PAF challenge in the guinea pig, SRI 63-675 inhibited the hemoconcentration (ED50 = 17 micrograms kg-1 i.v.) and bronchoconstriction (ED50 = 24 micrograms kg-1 i.v.) responses. In the primate, the ED50 was 28 micrograms kg-1 i.v. against 3.5 micrograms kg-1 PAF-induced hemoconcentration. The ratio (1:6) in the primate of PAF used (6.3 nmol kg-1) to antagonist at the ED50 (40.7 nmol kg-1) indicates exceptional potency of SRI 63 675 in this species. The inhibition by SRI 63-675 of the major PAF-induced effects in the rat, guinea pig and primate suggests a common receptor may be involved in the expression of these PAF responses. PMID- 3037719 TI - Dipyridamole inhibits leukotriene B4 synthesis. PMID- 3037720 TI - Endogenous forms of fibrinogen in Hep G2 cells. AB - The three polypeptide chains of fibrinogen, A alpha, B beta and gamma chain, are synthesized on separate polysomes. Fully formed fibrinogen is a six chain, disulfide-linked, dimeric molecule with a molecular weight of 340kDa. Previous pulse-chase studies with L-35 S methionine using the human hepatocellular carcinoma cell line, Hep-G2, showed that the three chains are not immediately disulfide-linked and that there exist intermediate precursors as well as pools of A alpha and gamma chains (J. Biol. Chem. 259, 10574-10581, 1984). In this study the endogenous levels of fibrinogen and its precursors are measured by two different methods; pulse and steady-state labelling with L-35 S-methionine and immunoblotting. In Hep-G2 cells intracellular fibrinogen-related antigen is primarily (30-53%) composed of an A alpha-gamma complex and, to a smaller degree, of fully-formed fibrinogen (13-33%). Furthermore, the Hep G2 cell also contains endogenous pools of free gamma chain (11-26%). Other fibrinogen precursors (namely, the B beta-A alpha, B beta-gamma complexes as well as the fibrinogen half-molecule) do not appear to accumulate intracellularly. Most, if not all, of these precursors occur as isoforms but this heterogeneity is not due to varying degrees of glycosylation. In all the intracellular fibrinogen forms identified thus far, free sulfhydryl groups, detected by 14C-iodoacetamide incorporation, occur only in the A alpha-gamma complex and the free gamma chains. PMID- 3037721 TI - Antithrombotic effect of some platelet modifying drugs. PMID- 3037722 TI - Inhibition of von Willebrand factor binding to platelets by two recognition site peptides: the pentadecapeptide of the carboxy terminus of the fibrinogen gamma chain and the tetrapeptide arg-gly-asp-ser. AB - When platelets are stimulated by thrombin or ADP, von Willebrand factor (vWf) binds to the platelet glycoprotein IIb/IIIa (GPIIb/IIIa). Two recognition sites for vWf binding have been identified on the GPIIb/IIIa. One is a tetrapeptide, arg-gly-asp-ser (RGDS), and the second is a dodecapeptide with the sequence HHLGGAKQAGDV. We used these purified peptides to analyze the recognition specificities for the platelet binding of vWf. The RGDS and the pentadecapeptide totally inhibited 125I-vWf in a dose-dependent manner. The IC50 was 8 microM for the RGDS and 40 microM for the pentadecapeptide. These results indicate that the RGDS binds with higher affinity to a vWf binding site on GPIIb/IIIa than does the pentadecapeptide site. When the two peptides were incubated together, they augmented the inhibition of vWf binding. These data support the evidence that the GPIIb/IIIa has two distinct binding sites for vWf binding. The RGDS site appears to have a higher affinity for the vWf binding site than does the pentadecapeptide site. PMID- 3037723 TI - [An important source of error in Rh(D) typing]. PMID- 3037724 TI - [Polyneuropathy in gasoline workers]. PMID- 3037725 TI - [Epidemic of erythema infectiosum in Norway 1984-1986]. PMID- 3037726 TI - The distribution of HLA-A, B, C, DR antigens in Chinese patients with hepatocellular carcinoma in Taiwan. AB - To evaluate the role of genetic factor in hepatocellular carcinoma, HLA-A, B, C and DR locus antigens were typed in 170 Chinese patients. Forty-three HLA-A, B, C, DR-related antigens were assayed by the conventional method. No significant risk antigen can be identified among these antigens. The distribution of each antigen was not related to sex. HBsAg, and alpha-fetoprotein status. The results suggest that there are no specific HLA-A, B, C and DR antigens patterns or frequencies associated with the development of hepatocellular carcinoma. PMID- 3037727 TI - Influence of cadmium on the metabolism of vitamin D3 in rats. AB - In order to obtain further information on the effects of cadmium (Cd) on the mechanism of activation of vitamin D3 in the kidney, the serum concentrations of 1,25-dihydroxycholecalciferol [1,25(OH)2D3] and 24,25-dihydroxycholecalciferol [24,25(OH)2D3] were determined by radioimmunoassay techniques. The serum concentration of 1,25(OH)2D3 in Cd-exposed rats was always higher than that in control rats. The concentrations of serum 1,25(OH)2D3 in parathyroidectomized rats (PTX), both in the control and in the Cd-exposed groups, were markedly lower than those in non-PTX rats. On the other hand, the concentration of serum 24,25(OH)2D3 in Cd-exposed rats was less than that in control rats. In other words, the pathway from secretion of parathyroid hormone (PTH) to synthesis of 1,25(OH)2D3 or 24,25(OH)2D3 was normal, but the pathway from stimulation of PTH to secretion of PTH was abnormal. These results suggest that rats exposed to Cd for 90 days were in a state of either deficiency of 1,25(OH)2D3 or excess secretion of parathyroid hormone. Although hypophosphatemia occurred in the Cd exposed rats, the 1,25(OH)2D3 serum level in rats was not increased by PTX. On the basis of these results, it is suggested that hypophosphatemia occurring after the exposure of rats to Cd is a secondary hypophosphatemia. PMID- 3037728 TI - An in vitro model for studying the toxicity of 2,3,7,8-tetrachlorodibenzo-p dioxin to human thymus. AB - A coculture system of human thymic epithelial (HuTE) cells and thymocytes (T lymphocyte precursors) has been established and characterized as an in vitro model for assessing the potential toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) to human thymus. HuTE cells in culture were responsive to TCDD as judged by induction of the cytochrome P1-450 monooxygenase activities, 7-ethoxycoumarin O-deethylase (ECOD) and 7-ethoxyresorufin O-deethylase (EROD). Measurement of the responsiveness of thymocytes cocultured on TCDD-pretreated HuTE monolayers to the mitogens concanavalin A (Con A) and phytohemagglutinin (PHA) indicated that TCDD can act directly on HuTE cells to suppress thymocyte maturation (at a concentration of 10 nM, TCDD produced a 25 to 50% inhibition of thymocyte responsiveness to Con A and PHA). Both the induction of cytochrome P1-450 monooxygenase activity (EC50 values approximately 1 nM) and immunosuppressive responses elicited by TCDD in HuTE cells were concentration-dependent and stereospecific (as judged by the relative activities of chlorinated dibenzo-p dioxin and dibenzofuran isomers), indicating involvement of the Ah receptor which was detected in all HuTE strains examined. Initial characterization of these Ah receptor-mediated responses in several strains of HuTE cells indicated marked interstrain differences in maximally inducible ECOD and EROD activities which did not appear to directly correlate with measured concentrations of the cytosolic Ah receptor, and in certain strains examined, differences in sensitivity and magnitude were observed for TCDD-evoked immunotoxic responses but not always for the induction response. These data on the actions of TCDD on cultured HuTE cells suggest that human thymus is a target for TCDD and related halogenated aromatic compounds. In HuTE cells, measurement of either the Ah receptor concentration or of marker responses such as the induction of cytochrome P1-450 alone cannot provide an accurate quantitative assessment of susceptibility to TCDD-induced thymus toxicity. PMID- 3037729 TI - In vivo exposure to four ellipticine derivatives with topoisomerase inhibitory activity results in chromosome clumping and sister chromatid exchange in murine bone marrow cells. AB - A single dose of 9-hydroxy-ellipticine, 2-N-methyl-9-hydroxy-ellipticine, 9 fluoro-ellipticine, and 9-amino-ellipticine (5 to 10 mg/kg body wt, ip) resulted in murine bone marrow toxicity as shown by chromosome clumping, chromatid aberrations, and micronuclei formation. An increase in sister chromatid exchanges (SCE) was also observed. These effects are most likely directly related to the topoisomerase inhibitory effect of these drugs since topoisomerase II is involved in the separation of intertwined chromosomal DNA molecules during mitosis as well as being a mediator of DNA exchanges. The two antitumor drugs 2-N-methyl-9 hydroxy-ellipticine and 9-hydroxy-ellipticine were most genotoxic with chromosome abnormalities occurring in 33-95% of the cells and SCE on the order of 12.3 to 19.2 events per cell. Both of these drugs show high topoisomerase II inhibitory activity in vitro. In contrast, 9-amino-ellipticine and 9-fluoro-ellipticine were less genotoxic with chromosomal abnormalities occurring in 14-17% of the cells and SCE on the order of 7.1 to 7.7 events per cell. These two derivatives are both inactive toward experimental tumors and show less topoisomerase II inhibitory activity in vitro. Our results suggest that the ellipticine drugs owe at least some of their cytotoxicity to their genotoxic effects, which seem to be mediated through interaction with topoisomerase II. PMID- 3037730 TI - Effect of Cd administration on the pituitary-adrenal axis. AB - The possibility that Cd administration could activate the pituitary-adrenal axis has been studied. It was found that acute administration of Cd strongly increased serum corticosterone levels. Adrenocortical activation correlated well with serum Cd levels, suggesting that adrenal activation was dependent on the presence of the metal in the blood. In addition, our data clearly demonstrate that increased corticosterone secretion caused by Cd was, at least in part, mediated by adrenocorticotropin release. Therefore, it appears that Cd might cause similar endocrine and metabolic changes as classical stress agents. PMID- 3037731 TI - Enhanced chemotaxis and superoxide anion production by polymorphonuclear leukocytes from nicotine-treated and smoke-exposed rats. AB - Although previous studies have shown that polymorphonuclear leukocytes (PMNs) exposed to nicotine in vitro exhibit enhanced superoxide anion generation and chemotactic responses, it is not known whether in vivo exposure to the alkaloid causes the same alterations in PMN function. Accordingly, this study evaluated superoxide anion generation evoked by phorbol myristate acetate (PMA) and chemotactic responses to formylmethionylleucylphenylalanine (fMLP) in PMNs isolated from rats treated acutely or subchronically with nicotine and from rats chronically exposed to cigarette smoke. Acute or subchronic (twice daily for 7 days) i.p. injection of 0.2 or 0.02 mg/kg nicotine potentiated PMA-induced superoxide anion generation by PMNs. Similarly, acute i.p. injection of 0.2 mg/kg nicotine or subchronic treatment with 0.02 mg/kg nicotine potentiated fMLP induced chemotaxis. Subchronic treatment with 0.2 mg/kg of the alkaloid blunted fMLP-induced chemotaxis, in contrast to the potentiating actions of the lower dose. Treatment with nicotine mimicked the effects of tobacco smoke exposure. A 15-week exposure regimen to either sidestream and mainstream smoke from University of Kentucky 2R1 reference cigarettes potentiated PMA-induced superoxide anion generation. Mainstream but not sidestream smoke also enhanced chemotactic responses to fMLP. Viewed collectively, these observations indicate that in vivo exposure to nicotine or to tobacco smoke augment PMN superoxide anion generation and chemotactic responses to selected stimuli and thus implicate such adverse actions of smoking on PMN function in certain pathologies associated with excessive tobacco smoke exposure. PMID- 3037732 TI - Effect of food intake on blood lead concentration in workers occupationally exposed to lead. AB - Results from a cross-sectional study showed the concentration of lead in the blood of male workers, aged 20-55 years, occupationally exposed to lead in a steel factory, to be negatively correlated with the daily nutritional content of dietary fiber, iron and vitamin B1 (thiamine) intake. Furthermore, in experiments with rats injected subcutaneously with lead acetate, lead levels in blood and femur of animals on a vitamin-rich laboratory chow were lower than those fed a general laboratory chow. Moreover, in the group fed the vitamin-rich chow, lead excretion in feces increased, while excretion in urine did not. These results suggest that lead excretion from the body may be increased by a high intake of nutrients such as thiamine, iron and fiber, that lead excretion in feces via bile may be enhanced by a large intake of vitamins such as thiamine and that accordingly the lead concentration in the blood of the workers is reduced. PMID- 3037733 TI - Inhibition of rat brain cytochrome oxidase activity by pyrolysed products of methyl isocyanate. AB - The effects were studied of methyl isocyanate (MIC) and its thermally degraded products (dMIC) on rat brain cytochrome oxidase activity. Pure MIC did not inhibit brain cytochrome oxidase activity. A significant inhibition of brain cytochrome oxidase activity by dMIC was observed both in vivo and in vitro. The presence of cyanide in pyrolysed products of MIC has also been confirmed by chemical methods. PMID- 3037734 TI - Helminth-induced intestinal inflammation. AB - Gastrointestinal inflammation is a prominent feature of protective reactions in animals immune against helminths. Infiltration into the inflamed mucosa of various cells and their subsequent activation result in the elaboration of an array of pharmacologically and biologically active substances. The release of mediators is also associated with alterations in the epithelial layer. Furthermore, increased smooth muscle reactivity and enhanced secretory function of the mucosal tissue contribute to the development of an unfavourable environment and lead to worm expulsion. Mediators elaborated from inflammatory cells, whether associated with cell granules (i.e., preformed) or de novo generated from membrane phospholipids, possess a number of potent vasoactive and spasmogenic properties which may contribute to events leading to worm elimination. The lipoxygenase metabolites of arachidonic acid (leukotrienes) derived from cell membranes probably contribute to the state of intestinal hypersensitivity against helminths. The measurement of elevated levels of these lipid mediators following worm challenge of immune, but not control, rats suggests that leukotrienes may play a role in amplifying and augmenting the inflammatory process associated with worm expulsion. PMID- 3037735 TI - Mucosal leishmaniasis ("espundia" Escomel, 1911). AB - One of the more serious clinical forms of leishmaniasis occurs in espundia when the mucosae of the upper respiratory passages are inflamed. This complication is a metastasis from a skin lesion caused by Leishmania braziliensis braziliensis (Lbb) although cases have been described associated with other leishmanial species. Epidemiological data suggest that a detectable mucosal metastasis occurs in fewer than 5% of patients infected with Lbb in our study area. The determinants of this complication are still largely obscure. The granuloma usually commences on the nasal septum. In about two-thirds of our patients the lesion remained restricted to the nose. In the rest the pharynx, palate, larynx and lips were involved, in this order. It is often difficult to isolate the parasite and for routine diagnosis the leishmanin skin reaction and serological tests are helpful. Although a serious condition, with possible mutilation and even death as subsequent complications, treatment is still mainly with pentavalent antimonials, introduced 40 years ago. These are most unsatisfactory for field use, being given parenterally and relatively toxic. In mucosal leishmaniasis, if sufficient antimony can be administered in a regular daily dose, the relapse rate is small (3 of 42 patients followed for a mean of 5 years). Also, antimony treatment of the initial skin ulcer due to Lbb followed for a mean of 4 years of 83 patients resulted in subsequent mucosal metastasis in only 2. Since espundia is relatively rare, specific treatment targeted to this specific problem is the efficient short term solution. At present there is no satisfactory alternative drug to those in current use. PMID- 3037736 TI - Relationship of some biochemical characteristics of Staphylococcus aureus to enterotoxin production. AB - A total of 151 enterotoxigenic and 98 non-enterotoxigenic strains of Staphylococcus aureus were examined for production of coagulase, phosphatase, deoxyribonuclease, haemolytic activity, glucose and mannitol fermentation, typical growth on Baird-Parker medium, pigment production and growth under anaerobic conditions. Enterotoxigenic strains showed a higher tendency to demonstrate haemolytic activity, but none of the other biochemical properties could be correlated with the ability of the staphylococci to produce enterotoxins. It appears that other ancillary tests must be developed to be useful in the differentiation of enterotoxigenic and non-enterotoxigenic staphylococci. PMID- 3037737 TI - Cytomegalovirus infection after renal transplantation. Pulmonary dysfunction measured by decreased diffusing capacity for carbon monoxide in patients with symptomatic and asymptomatic infection. PMID- 3037738 TI - Acute transverse myelitis during disseminated cytomegalovirus infection in a renal transplant recipient. PMID- 3037739 TI - Successful therapy of Niemann-Pick disease by implantation of human amniotic membrane. AB - In a patient with a lysosomal storage disorder, not involving the CNS, repeated implantations of human amniotic sheets have proved to provide a successful approach to enzyme replacement therapy. Implantation of pure epithelial cells, separated from the other cell types of the amnion, might markedly improve the procedure, avoiding some risks of host-versus-graft rejection. PMID- 3037740 TI - Monkeypox virus in relation to the ecological features surrounding human settlements in Bumba zone, Zaire. AB - Since monkeypox virus was discovered in animals in 1958 and in man in 1971, several efforts have been made to identify the reservoir of the virus in nature. In July 1985 a study on human environments and animals suspected to maintain virus transmission was carried out in northern Zaire. The study revealed three well demarcated areas: the human settlement area, the agricultural area and the primary tropical rain forest. The first two were found inhabited by terrestrial and arboreal rodents and bats, while the larger animals have abandoned them. Data on human morbidity collected in the preceding years suggested that most of the patients were infected either in the settlement or in the agricultural area. Isolation of the virus from a squirrel confirmed this suggestion and proved, on the other hand, that the virus could circulate in an area deprived of the larger wild animals, including primates. The virus isolation and results of serologic testing suggest that out of three groups of animals inhabiting the agricultural area, the squirrels Funisciurus anerythrus should be considered a priority candidate to sustain virus transmission. PMID- 3037741 TI - [Genetic transformation of somatic cells. XIII. Limited replication of DNA containing a fragment of human satellite DNA III in mouse cells]. AB - The plasmids containing the variant sequence of human satellite III "rescued" after replication in Chinese hamster cells were transfected into Chinese hamster, mouse and human cells by DEAE-dextran method. Several days after transfection extrachromosomal fractions were isolated, treated with DpnI, transformed into E. coli. In mouse cells, transformed with oncogene v-myc, after transient transfection of HS3-containing plasmids the appearance of rearranged and non rearranged DpnI-resistant plasmids has been found. At the same time MboI sensitive plasmids were not found in this material. The data suggest a limited replication (1 round) of HS3-containing plasmids in mouse cells transformed with oncogene v-myc. PMID- 3037742 TI - Challenge study on infectious bursal disease in chicks derived from vaccinated hens. AB - Presence and levels of maternal antibody (MA) in broiler chicks derived from hens vaccinated with a live infectious bursal disease (IBD) vaccine were investigated by a quantitative agar-gel precipitin test. At day old 100% of the chicks tested had MA; by 17 days of age it was present in only 10%. The mean MA level at day old was 337.5 UK units/ml but decreased to 6.3 UK units/ml at 17 days of age. Randomly selected chicks from the pool studied were challenged at weekly intervals from day old for 29 days with an IBD virus obtained from a natural outbreak. Subclinical and clinical disease were observed in chicks challenged at eight and 29 days of age respectively. PMID- 3037743 TI - First isolation of IBR virus in Turkey. PMID- 3037744 TI - Methotrexate with citrovorum factor in low-risk gestational trophoblastic tumor. AB - From January 1976 through December 1985, methotrexate (MTX) with citrovorum factor (CF) was administered as primary treatment to 57 patients with low-risk gestational trophoblastic tumor (GTT); 51 patients were non-metastatic and 6 were metastatic GTT. The median number of courses needed to achieve biochemical remission was two (range, 1-7). Complete remission was attained in 95% of non metastatic GTT patients with postmolar persistent trophoblastic disease, but when choriocarcinoma was histologically confirmed, this fell to 60%. The cure rate of metastatic GTT patients was only 50%. The overall remission rate with the MTX-CF combination was 84.2%. Toxicity was mild, consisting of myelosuppression and mucositis. Fifteen patients were resistant to MTX-CF, or relapsed subsequently, but they all achieved remission with chemotherapy rescue treatment (VP 16 alone, EMA/CO, CHAMOCA). Two patients required a pulmonary lobectomy. They are all still alive in biochemical remission with a median survival of 54 months. Our experience suggests that drug resistance and relapse rate seem related to a beta HCG value higher than 10(4), an enlarged uterus with myometrial deep involvement, and a histologically confirmed diagnosis of choriocarcinoma. In conclusion, the MTX-CF combination is effective in postmolar GTT, whereas a different therapeutic approach may be considered for a "special" low-risk group of patients, on the basis of prognostic factors. PMID- 3037745 TI - [Changes in the levels of various substrates of nitrogen metabolism and tricarboxylic acid cycle during experimental acidosis in calves]. AB - The effect of experimental metabolic acidosis and its correction for nitrogen and energy metabolism was studied in new-born calves. It was discovered that a change in the acid-base balance towards acidosis causes a sharp increase in "ammoniogenesis", urea formation and inhibition of the tricarboxylic acid cycle, which is also observed in calves suffering from dyspepsia with symptoms of acute diarrhea. Alongside with the use of therapeutic measures for treating dyspepsia of new-born calves, it is necessary to control the acid-base balance of blood in the calves and in case of revealing the acidosis state to use means of its correction. PMID- 3037746 TI - [Effect of combined anesthetics on the activity of various myocardium enzymes]. AB - It is demonstrated by experiments with rabbits that the Ca2+-ATP-ase activity is stabilized when using combined anesthetics (diacetylcholine + halothane + N2O) as distinct from application of halothane. A decrease in the cholinesterase activity is less pronounced than under the halothane action but more than with the diacetylcholine application. A decrease in the Na+, K+-ATP-ase activity is observed with all types of anesthesia. A considerable inhibition of creatine kinase under the action of combined anesthesia and halothane and an increase of the lactate dehydrogenase activity under diacetylcholine application in mitochondria are shown. Reliable differences in the succinic dehydrogenase activity are not detected. PMID- 3037747 TI - [The role of adrenoreceptors in the mechanism of pharmacological action of cavinton]. AB - Having studied the functional state of beta-adrenoreceptors in vitro by the radioligand (3H-dihydroalprenalol) method it has been shown that cavinton in the concentration of 2.5 X 10(-6) M considerably increases the affinity of beta adrenoreceptors for the ligand. In the in vivo experiments on rats the intraperitoneal injection of cavinton in the dose of 2 mg/kg causes an increase in the affinity of synaptosomal beta-adrenoreceptors for the ligand 10 days after chronic administration without altering the concentration of the receptors (Bmax) themselves. 20 days after the injection of the drug the affinity still grows and this effect of cavinton is retained even 30 days after the injection. PMID- 3037748 TI - [Breast metastases as the initial symptom of extramammary carcinomas]. PMID- 3037749 TI - [Cellular regulatory systems in relation to stimulus-response coupling]. PMID- 3037750 TI - Tumor thrombosis of the inferior vena cava due to retroperitoneal germ cell tumor. AB - Three patients (2 males and 1 female), aged 17-28 years, with tumor thrombosis of the inferior vena cava due to retroperitoneal germ cell tumor are reported. Diagnostic and therapeutic implications of this condition are discussed. PMID- 3037751 TI - A multi-substrate metalloproteinase from rheumatoid synovial cells. PMID- 3037752 TI - Atrial peptides inhibit Na+ entry-dependent oxygen consumption in rabbit inner medullary collecting duct cells. PMID- 3037753 TI - 3-Hydroxykynurenine as a possible mechanism of epileptic seizures associated with neonatal vitamin B6 deficiency. PMID- 3037754 TI - Recognition of hepatitis B surface antigen by the human hepatoma cell line HepG2. PMID- 3037755 TI - Prognostic factors in patients with carcinomas of the testicle, prostate, and bladder. PMID- 3037756 TI - [The effect of colostral immunity on the active formation of antibodies in pigs after the administration of inactivated vaccine against Aujeszky's disease]. AB - In a large herd of pigs where a trial was performed to cure the animals from Aujeszky's disease (AD) by applying to all animals an inactivated vaccine, a post vaccination antibody response was studied in piglets coming from the sows that were vaccinated several times. When the piglets were vaccinated at the age of eight weeks (the average virus-neutralizing titer (VNT) of colostral antibodies was 1:11.4) and revaccinated at the age of 11 weeks, 73% of the forty-five animals (examined at the age of 17 weeks) did not have any virus-neutralizing (VN) antibodies in the blood serum. After the third vaccination dose (at the age of 17 weeks), 11% of piglets did not have any VN antibodies if they were examined at the age of 22 weeks (the average antibody VNT was 1:15.3). Applying the ELISA procedure, the antibodies were demonstrated in the sera of all piglets after three vaccination doses. Shifting the time intervals of vaccination (at the age of 8, 13 and 19 weeks), the VN antibodies were found out after three vaccination doses in the sera of all piglets examined at the age of 23 weeks (the average VNT was 1:56.4). After three vaccination doses at the age of 12, 17 and 23 weeks, the VN antibodies were also demonstrated in all piglets at the age of 27 weeks (the average VNT was 1:208). PMID- 3037757 TI - AIDS and maedi-visna: parallels and divergences. PMID- 3037758 TI - Pathogenesis of maedi-visna. AB - In most cases, maedi-visna virus infection is characterised by a subclinical, persistent virus-carrier state. However, in heavily infected flocks, economically significant disease does occur, mainly apparent as ill-thrift and chronic respiratory disease (maedi) in older ewes and as an indurative mastitis, which can result in delayed weight gain of suckled lambs. Meningoencephalitis (visna) and arthritis may also occur. Maedi-visna virus, a lentivirus, replicates via proviral intermediary DNA copies of its RNA genome in circulating monocytes, in which replication is highly restricted, and in tissue macrophages, where viral genome expression is more evident. The presence of macrophages expressing viral antigens on their surface in lungs, udder, joints or central nervous system tissues provides a focus for a local mononuclear cell inflammatory response. Factors which may contribute to macrophage activation and the development of the inflammatory response are discussed in the context of virus replication, transmission of infection and disease susceptibility. PMID- 3037760 TI - Caprine arthritis-encephalitis in France. PMID- 3037759 TI - Mixed infection with Chlamydia psittaci, fowlpox virus and Haemophilus gallinarum in broiler breeder chicks. PMID- 3037761 TI - Survival of bovine virus diarrhoea virus in blood from persistently infected cattle. PMID- 3037762 TI - Caprine arthritis-encephalitis in the Basque country, Spain. AB - A goat herd severely affected by arthritis was studied. The most representative clinical signs consisted of articular swelling, mainly of the carpal joints, and the subsequent locomotor disorders. Some goats also showed signs of central nervous system involvement. Examinations of joint fluid revealed an increased number of mononuclear blood cells, mostly lymphocytes. Gross and microscopic articular lesions were of inflammatory and degenerative types. Periarticular connective tissue, synovial bursae, tendons and tendon sheaths were predominantly affected. Inflammatory lesions were those of a chronic hyperplastic tenosynovitis with fibrosis of the connective tissue components. Degenerative changes consisted mainly of necrosis and mineralisation of articular-related structures. Histological lesions in the central nervous system were those of a nonpurulent encephalitis initially located in periventricular areas, but in one case extensive encephalomalacia was also seen. Of the 80 animals sampled 82.5 per cent showed seropositive reactions against an ovine progressive pneumonia virus antigen. None was seropositive to brucella and titres to chlamydia were low. Attempts to isolate chlamydia and mycoplasma from affected joints and several organs failed. Different bacteria were recovered from a few samples but did not seem significant. Syncytium-forming viral particles were isolated from several organs, mainly the lungs, synovial membranes and lymphoid tissue of almost all the slaughtered animals. These particles were identified as lentiviruses by electron microscopy. The clinical signs, lesions serological results and microbiological findings, led to a diagnosis of caprine arthritis-encephalitis. This syndrome has not been recognised in Spain previously. PMID- 3037763 TI - Hypoglycaemia due to a functional pancreatic islet cell tumour (insulinoma) in a ferret (Mustela putorius furo). PMID- 3037765 TI - Comparison by the neutralisation assay of pairs of non-cytopathogenic and cytopathogenic strains of bovine virus diarrhoea virus isolated from cases of mucosal disease. AB - Neutralising antibody to non-cytopathogenic and cytopathogenic strains of bovine virus diarrhoea virus (BVDV) was assayed in a microtitre test in which cultures of calf testis cells were stained by the immunoperoxidase method to detect viral replication. Fourteen BVDV strains were compared in cross neutralisation tests with antisera prepared in gnotobiotic calves. Ten of the strains comprised five pairs of non-cytopathogenic and cytopathogenic BVDV. Each pair was isolated from an animal with mucosal disease. All five animals were from five separate outbreaks of the disease. Each pair of strains from the same outbreak was found to be antigenically indistinguishable. In contrast, when the coefficient of antigenic similarity was calculated 11 of 45 comparisons between the pairs and 46 of 91 comparisons between all 14 viruses gave R values that distinguished strains. The observations suggest that an antigenic spectrum within a single related group exists for BVDV strains, rather than distinct serotypes. The findings are also consistent with the suggestion that cytopathogenic strains from natural outbreaks of mucosal disease arise by mutation from non-cytopathogenic virus. PMID- 3037764 TI - Assessment of a general therapeutic protocol for the treatment of acute T-2 toxicosis in swine. AB - T-2 toxin, a trichothecene mycotoxin suspected of being used as a chemical warfare agent, was administered iv to swine at a dose of 3.6 mg/kg body weight (iv LD50 approximately 1.2 mg/kg). Four different therapeutic protocols were assessed for their efficacy in the treatment of the resultant acute T-2 toxicosis syndrome. One therapeutic protocol included the combined use of metoclopramide, activated charcoal, magnesium sulfate, dexamethasone sodium phosphate, sodium bicarbonate and normal saline. The other 3 protocols utilized the same agents less 1 of the following: the combination of activated charcoal and magnesium sulfate, sodium bicarbonate, or normal saline. All 4 treatment groups showed improved survival times compared to a positive T-2 control group. Within the limits of the study, it would appear that the removal of activated charcoal and magnesium sulfate was most detrimental to the T-2 toxin-dosed swine. PMID- 3037766 TI - The propagation of feline panleukopenia virus in micro-carrier cell culture and use of the inactivated virus in the protection of mink against viral enteritis. AB - Using microcarrier cell culture for the production of virus antigen, a formalin inactivated feline panleukopenia virus vaccine was evaluated for protection of mink against specific mink enteritis virus infection. The vaccine showed a good immunogenic effect in mink when used either alone or in combination with Clostridium botulinum type C-toxoid and/or Pseudomonas aeruginosa vaccine. A single vaccination induced persistent immune responses for periods of at least 1 year, as evaluated by ELISA and challenge tests. Neither immunological interference between vaccine constituents nor adverse reactions were observed. PMID- 3037767 TI - A sensitive plaque assay for bovine rotavirus in cultures of the bovine cell line GBK. AB - The Lincoln strain of bovine rotavirus was found to replicate with cytopathic effects in cultures of GBK cells, a stable cell line derived from bovine kidney, when the cultures were maintained in the presence of trypsin. The virus was readily passaged and the infected cells were shown to contain specific viral antigen by indirect immunofluorescent staining. The virus formed plaques in GBK cell monolayers, when trypsin was incorporated in the agar overlay medium. The plaque count increased about twofold when diethylaminoethyl dextran was further included in the overlay medium. Plaque assay in GBK cells was more sensitive than that in MA-104 cells previously reported by Matsuno et al. The specificity of plaques was confirmed by specific inhibition with antiserum against the Lincoln strain. PMID- 3037768 TI - Bovine viral diarrhea virus-infected MDBK monolayer as antigen in enzyme-linked immunosorbent assay (ELISA) for the measurement of antibodies in bovine sera. AB - A specific ELISA for the detection of IgG antibodies against bovine viral diarrhea virus (BVDV) has been developed and was compared with the seroneutralization (SN) titers for a total of 60 bovine serum samples. A BVDV infected Madin-Darby Bovine Kidney (MDBK) cell monolayer served as test antigen. The following results were obtained: a coefficient of correlation (r) of 0.63 (P less than 0.01) between BVDV-ELISA and SN titer in BVDV-seroconverted animals and 0.71 for all sera, including that from BVDV vaccinated animals; a sensitivity of 92%, a specificity of 91%, a concordance of 92%; and a demonstration of a parallel increase in BVDV-ELISA and SN titers in animals with seroconversion. The BVDV-ELISA permits a more complete evaluation of the humoral immune response to BVDV. PMID- 3037769 TI - Cell proliferation and differentiation in intramucosal and advanced signet ring cell carcinomas of the human stomach. AB - In order to study the progression of signet ring cell carcinomas in the human stomach, we compared cell proliferation and differentiation between small and large intramucosal cancers, and between intramucosal and advanced cancers. Fine structurally, signet ring cells were differentiated to 3 cell types: a foveolar, a glandular and an intestinal type. In the mucosa, the foveolar-type cells and glandular-type cells were distributed at the superficial and the deep zone, respectively. In the small mucosal cancers, intestinal-type cells were rare and a layered structure was often seen. In this structure, the mode of cell production resembled that in the normal gastric mucosa; the foveolar-type signet ring cells in the superficial layer were not proliferative and the proliferating cells were small cells in the middle layer and a few glandular-type cells in the deep layer. In the large mucosal and advanced cancers, intestinal-type cells and proliferating small round cells were often distributed throughout the depth of the mucosa, and signet ring cells of the foveolar type were also proliferative. These findings indicated that large part of the signet ring cell carcinomas initially form the layered structure and that it becomes indistinct while intestinal-type cells appear as the tumour grows. However, we found several small advanced cancers, lacking both the layered structure and the intestinal-type cells. These cancers appear to start without the layered structure and progress very rapidly. PMID- 3037770 TI - Expression of the C-terminal peptide of human pro-bombesin in 361 lung endocrine tumours, a reliable marker and possible prognostic indicator for small cell carcinoma. AB - Small cell carcinoma of the lung is a highly malignant tumour. Its known biological products which include bombesin, do not allow the prediction of tumour behaviour. Molecular biology has revealed the amino acid sequence of human pro bombesin, which consists of a signal peptide, the bioactive bombesin molecule and a C-terminal peptide. We have raised a rabbit antiserum to the first (N-terminal) 21 amino acids of the predicted C-terminal peptide. A total of 505 (361 neuroendocrine) surgically resected pulmonary tumours were evaluated for the presence of immunoreactive bombesin and C-terminal peptide. Strong immunostaining was obtained with the antiserum to the C-terminal peptide of human pro-bombesin in 70% of the small cell carcinomas (175/250), in 63% of atypical (aggressive) carcinoids (31/49) but only in 16% of benign carcinoids (10/62). In contrast, bombesin immunostaining was focal and only moderately strong and the relative proportion of positive cases was quite evenly distributed amongst the neuroendocrine tumours: 35% of carcinoids (22/62), 22% of atypical carcinoids (11/49) and 25% of small cell carcinoma (62/250). None of the squamous, adeno, or large cell undifferentiated carcinomas were immunoreactive for bombesin or the C terminal peptide. Radioimmunoassay and chromatography of extracts of tumours recovered from wax blocks revealed high concentrations of C-terminal peptide immunoreactivity (241 +/- 66 pmol/g of tissue) in all 12 small cell carcinomas studied, moderate concentrations in carcinoid tumours (50 +/- 7 pmol/g) and none in non-small cell carcinomas. Patients with tumours showing immunoreactivity to the C-terminal peptide of human pro-bombesin had a significantly shorter survival time than those without immunoreactive peptide (185 +/- 16.49 days, mean +/- SEM, and with 1128 +/- 226 days, respectively P greater than 0.02). The apparent presence of the C-terminal peptide of human pro-bombesin in higher concentrations than bombesin in the more malignant class of endocrine tumours, mainly small cell carcinomas associated with the poorest prognosis, suggests that the antiserum to this C-terminal peptide is not only a useful pathological marker but may prove to be of value in investigating the biological behaviour of small cell carcinomas and predicting the clinical course of the disease. PMID- 3037771 TI - Human papillomavirus type 13 and focal epithelial hyperplasia of the oral mucosa: DNA hybridization on paraffin-embedded specimens. AB - 16 cases of focal epithelial hyperplasia (Heck's disease) were studied for the presence of human papillomavirus DNA by means of nucleic acid hybridization. Hybridization was carried out in situ with biotin-labelled probes of HPV 1, 6, 11, 13, 16, and 18 DNA under stringent and non-stringent conditions. Under non stringent conditions, 6 of 16 cases (38%) hybridized to a mixture of HPV 1, 6, 11, 16, and 18 DNA. When these probes were applied under stringent conditions, only one case could be shown to be weakly positive for HPV 6/11 DNA. Further stringent hybridizations, which were conducted with a HPV 13 probe on 12 of our 16 cases, revealed a positive result in 9 of 12 cases (75%). The results of our study strongly substantiate the concept that HPV 13 or a closely related HPV type is associated with lesions morphologically presenting as focal epithelial hyperplasia. PMID- 3037772 TI - Cyclosporine A inhibits herpes simplex virus-induced cell fusion but not virus penetration into cells. AB - The effects of the immunosuppressive peptide cyclosporine A on virus-induced cell fusion and virus penetration into cell were studied. Cyclosporine totally inhibited polykaryocyte formation by a syncytial strain of HSV-1 (MP). In contrast, a number of other hydrophobic peptides which inhibit the replication of orthomyxo- and paramyxoviruses and which were less effective in inhibiting fusion in model systems had little effect on HSV-induced cell fusion. Virus replication as measured by the yield of infectious virus and by incorporation of [35S]methionine into viral proteins was unaffected by treating cells with cyclosporine. Cyclosporine did not quantitatively inhibit cell surface expression of HSV glycoproteins or plaque formation of syncytial or nonsyncytial virus strains. However, plaques formed by HSV-1 (MP) on cyclosporine-treated monolayers were smaller than those on untreated monolayers and were morphologically similar to plaques produced by the nonsyncytial HSV strain F. Surprisingly, cyclosporine did not inhibit penetration of HSV, a process also thought to involve membrane fusion. Therefore, HSV-1-induced cell fusion and penetration of virus into cells either proceed by mutually exclusive mechanisms or are differentially sensitive to cyclosporine. PMID- 3037773 TI - The genomic RNA of diabetogenic encephalomyocarditis virus: characterization and molecular cloning. AB - The RNA of a diabetogenic variant of encephalomyocarditis (EMC) virus (D variant, ifp- phenotype) and a nondiabetogenic variant of EMC virus (B variant, ifp+ phenotype) which were derived from the same virus stock (M strain) have been compared. The size of both genomes is estimated at 7.7 kb. The poly(C) tract of EMC-D is estimated at 144 bases, whereas that of EMC-B is 141 bases in length. The untranslated 5' terminal 103 nucleotides are identical for B and D with preservation of a stable terminal hairpin structure. The entire open reading frame of both variants has been cloned and the restriction maps of 12 different enzymes are identical. These maps were compared to a computer-generated restriction map of another strain of EMC virus which has been cloned and sequenced by Palmenberg et al. (1984, Nucleic Acids Res. 12, 2969-2985). Approximately 50% of the restriction sites of the B and D variants had similar locations in this strain of EMC. We conclude that significant genetic variation exists between the M strain (B and D variants) and the Palmenberg strain of EMC virus. However, the B and D variants are very similar at the molecular level and comparison of their nucleotide sequences will be necessary to reveal the basis for their different biological properties. PMID- 3037774 TI - Equine herpesvirus type 1 defective-interfering (DI) particle DNA structure: the central region of the inverted repeat is deleted from DI DNA. AB - The inverted repeat (IRs) component of the genome of equine herpesvirus type 1 (EHV-1) is an important region of structure and function. It is a major constituent of the DNA of EHV-1 defective-interfering (DI) particles which have been shown to mediate the coestablishment of oncogenic transformation and persistent infection of hamster embryo cells. In addition, the IRs encodes the single EHV-1 immediate early gene and the 31.5K very early protein. DNA sequences encompassing EHV-1 internal IRs and the joint between the long (L) and short (S) regions were subcloned into the plasmid vectors pBR322 and pUC12. A total of 22 subclones were derived, including six Sa/l subclones in pBR322 and 12 SmaI subclones in pUC12. Individual subclones were employed in Southern blot hybridizations to define subclone homology to repeated, unique, or heterogeneous (het) DNA sequences within the EHV-1 genome. These studies revealed that the EHV 1 het region is contained entirely within the unique long region of the viral genome and is separated from the L/S junction by approximately 1.8 MDa of completely unique DNA sequences. Furthermore, these IRs subclones were employed in blot hybridizations to analyze the integrity of IRs DNA sequences within the cloned DNA of EHV-1 DI particles. These analyses demonstrated that IRs DNA sequences present in DI DNA were extensively rearranged and contained major deletions (0.80-0.83 map units) which removed a large portion of the single EHV-1 immediate early gene (0.78-0.83 and 0.95-1.00 map units) located in the IRs. Thus, these data and those previous studies (R. P. Baumann et al., 1984, J. Virol. 50, 13-21; R. P. Baumann, J. Staczek, and D. J. O-Callaghan, 1986, Virology 153, 188-200) indicate that the major subunits of the DI DNA molecule are comprised of selected sequences from the IRs component and a highly conserved short sequence located at the terminus of the L region of the standard viral genome. PMID- 3037775 TI - Molecular cloning of infectious proviral genomes of bovine leukemia virus. AB - Covalently closed circular DNA molecules of bovine leukemia virus were cloned in the lambda phage vector lambda gtWES-lambda B and subsequently in the plasmid vector pUC12. Proviral DNAs of 8.3 kb which have one copy of a long terminal repeat and uniform restriction endonuclease sites were preferentially obtained. Four randomly selected clones were examined for their biological activities by DNA transfection experiments. The ovine embryonic cells transfected with these clones formed syncytia which represent the expression of BLV genes. The infectious virions could be recovered from the transfectants. PMID- 3037776 TI - Enhancement of Epstein-Barr virus membrane protein (LMP) expression by serum, TPA, or n-butyrate in latently infected Raji cells. AB - The BamHI Nhet region of the EBV DNA is known to code for two proteins. One is a membrane protein (LMP) with an apparent molecular weight of 60,000 on SDS-PAGE which is expressed in latently EBV infected cells. The second protein, so far unidentified, is presumably a late protein with a calculated molecular weight of 28,000 Da. Antisera against both proteins were generated by immunizing rabbits with either a fusion protein containing 155 amino acids of the C-terminus of LMP and a 37,000 mol wt piece of the bacterial anthranilate synthase or with a C terminal synthetic peptide of 7 amino acids. These sera reacted with a protein varying in size between 60,000 and 65,000 mol wt on SDS-PAGE, found in all cell lines harboring EBV. In addition, these sera identified a second protein with an apparent molecular weight of 49,000 on SDS-PAGE in B95-8, P3HR-1, and M-ABA cells, which is presumably identical with the 28,000-Da protein mentioned above. Furthermore, with these sera a positive cytoplasmic immunofluorescence in 1 to 10% of the cells was obtained, depending on the cell line examined. Analyzing the nonproducer Raji cell line, the number of immunofluorescence-positive cells and the amount of the 60,000 protein, as judged by immunoblotting, was rapidly increased by addition of fresh medium with 10% fetal calf serum as well as by the tumor promoter TPA or to an even higher extend by n-butyrate. The kinetics of induction reached a maximum 24 hr after addition of medium plus 10% fresh serum or TPA or n-butyrate and decreased after 24 to 48 hr. Since the induction of the EBV early antigen (EA) associated proteins by TPA or n-butyrate exhibits a diverse kinetic with a maximum at 72 hr, the regulation of the 60,000 protein synthesis appears to be different from known EA-associated proteins. PMID- 3037777 TI - The two segments of the infectious bursal disease virus genome are circularized by a 90,000-Da protein. AB - The genome of infectious bursal disease virus (IBDV) consists of two segments of double-stranded (ds)RNA with molecular weights of 2.2 X 10(6) and 1.9 X 10(6) Da, respectively. After treatment of IBDV particles with proteinase K in the presence of sodium dodecyl sulfate (SDS), linear dsRNA molecules are released from the virus particles. However, after heating of virus particles at 100 degrees for 3 min in 1.5% SDS, without the protease, dsRNA-protein complexes can be seen under the electron microscope: Knob-like proteinaceous structures are linked to the ends of the dsRNA molecules of either size class which are circularized to form individual rings. A 90,000-Da IBDV structural polypeptide, the only protein encoded by the smaller genome segment, has been demonstrated to remain firmly linked to the IBDV genome under these conditions. No functional data exist about this circularizing protein; it is a probable candidate for an RNA-dependent RNA polymerase or an assembly protein for the two dsRNA segments. At high particle concentrations, or when the preparations are allowed to stand for several hours before spreading, these complexes tend to aggregate to form flower-like structures. PMID- 3037778 TI - Transcriptional activity of the gibbon ape leukemia virus in the interleukin 2 gene of MLA 144 cells. AB - The gibbon ape leukemia virus (GALV)-infected T-cell line, MLA 144, constitutively makes the lymphokine, interleukin 2 (IL 2), without stimulation by antigen or mitogen. This line contains two GALV insertions in the IL-2 gene: one in the 3' untranslated region of the gene and one 1200 bp 5' to the gene. It is likely that one or both of these viral insertions is(are) involved in activation of IL-2 expression. We investigated the ability of sequences within the LTR of MLA 144 cells (GALV-MLA) to act as transcriptional elements and have demonstrated here the presence of cis-acting sequences in the GALV LTR capable of enhancing transcription of the GALV promoter as well as two heterologous promoters, SV40 early and IL-2. The results indicate that insertion of the enhancer element(s) alone is not sufficient to activate IL-2 expression but can enhance levels of IL 2 expressed from the activated gene. PMID- 3037779 TI - A new type of papillomavirus associated with cancer of the uterine cervix. AB - A new human papillomavirus (HPV) type was detected by low-stringency Southern blot hybridization analysis of DNA from an endocervical adenocarcinoma. The genomic DNA of the virus, which was obtained as two BamHI fragments of 3.75 and 4.1 kb, was molecularly cloned into lambda L47 and subsequently subcloned into pBR322 for further characterization. Hybridization studies demonstrated that these viral DNA isolates were only distantly related to other HPV and thus represented a new type of HPV, called HPV 35. A restriction enzyme map was prepared which allowed a comparison of the genetic organization of this HPV with that of HPV 6b; the results demonstrated collinearity of the HPV 35 genome with that of HPV 6b. Prevalence studies revealed that HPV 35 was present in 2 of 158 (1%) anogenital intraepithelial neoplasia and in 3 of 69 (4%) anogenital cancers. Thus HPV 35 is a low-prevalence human papillomavirus associated with anogenital intraepithelial neoplasia and cancer. PMID- 3037780 TI - A collaborative report: rhinoviruses--extension of the numbering system from 89 to 100. AB - To define the number of rhinovirus serotypes, cross neutralization tests and characterization studies were completed on 25 candidate prototype rhinoviruses submitted to a third phase of a collaborative program. Based on the results, 11 distinct prototype strains were designated and the numbering system was extended to include 100 rhinoviruses. In addition, recent evidence indicates that over 90% of rhinoviruses isolated in three areas of the country could be typed with antisera for rhinovirus types 1-89. PMID- 3037781 TI - A small open reading frame in pseudorabies virus and implications for evolutionary relationships between herpesviruses. AB - An open reading frame coding for an 11-kDa protein was located downstream from the gI gene of pseudorabies virus (PRV). This open reading frame is homologous to an open reading frame (US9) in an analogous position in herpes simplex virus and to an open reading frame (US1) in a different position in varicella zoster virus. The open reading frame encoding the 11-kDa protein is in a region known to be deleted in live attenuated vaccine strains of PRV. PMID- 3037782 TI - Structural alterations in the long terminal repeat of an acquired mouse mammary tumor virus provirus in a T-cell leukemia of DBA/2 mice. AB - ML, a transplantable T-cell leukemia of DBA/2 mice, expresses the gag and env gene products of the murine mammary tumor virus (MuMTV). Analysis of the genomic DNA of ML cells using the restriction enzyme HindIII and hybridization with MuMTV specific probes revealed that the ML cells contained two or more newly integrated MuMTV proviruses (ML-MuMTV). Further analysis of these proviruses with a combination of Mspl and Pstl enzymes showed that the long terminal repeat (LTR) (ML-MuMTV LTR) of the ML-MuMTV provirus(es) was structurally different from the LTRs of both exogenous and endogenous MuMTV proviruses of DBA/2 mice. In order to characterize the nature of the structural alterations in the ML-MuMTV LTR, we cloned a 4.0-kb HindIII fragment containing the 3' half of an acquired provirus. Sequence analysis of the ML-MuMTV LTR of this acquired provirus revealed a deletion of a 387-bp segment that maps between the 5' nucleotide 616 and the 3' nucleotide 1003 of the normal MuMTV LTR and duplication of a 102-bp fragment that mapped between 514 and 616. In addition to two point mutations in the direct repeat, the proviral ML-MuMTV LTR has also acquired 9- and 7-bp segments at the 5' and 3' sites of the duplicated 102-bp segment, respectively. Since direct repeats in the U3 regions of a number of LTRs have been found to be associated with enhancer function, we examined the enhancer function of the U3 region sequences of the ML-MuMTV LTR using enhancer-dependent transient expression assay of chloramphenicol acetyltransferase (CAT) gene in NIH 3T3 cells. Our studies have shown that the U3 region sequences of the rearranged ML-MuMTV LTR have the ability to enhance the expression of the CAT gene 12- to 15-fold more than the U3 region sequences from the normal MuMTV LTR. The presence of a direct repeat in the ML-MuMTV LTR and its ability to enhance the transcription of adjacent genes is analogous to the LTRs of certain murine leukemia viruses. PMID- 3037783 TI - Protection of cattle from bovine herpesvirus type I (BHV-1) infection by immunization with individual viral glycoproteins. AB - The major glycoproteins gI, gIII, and gIV of bovine herpesvirus-1 (BHV-1) were found to induce high levels of antibody in cattle which could neutralize virus and participate in antibody-dependent cell cytotoxicity of BHV-1-infected cells. Immunized animals were fully protected from disease, using a BHV-1/Pasteurella haemolytica aerosol challenge model but not from infection with the virus. Thus, virus could still replicate in the nasal passages of immunized animals, although to a lesser extent than in placebo-treated animals or animals immunized with a commercial killed whole virus vaccine. Systemic spread of the virus in immunized animals did not appear to occur since there was not a dramatic alteration of leukocyte function following challenge. These results suggest that any one of the three major BHV-1 glycoproteins may be useful as a subunit vaccine either individually or in combination. PMID- 3037784 TI - Multiple reiteration of a 40-bp nucleotide sequence in the inverted terminal repeat of the genome of a canine adenovirus. AB - The DNA of a vaccine strain of canine adenovirus type 1 [ICHV vaccine; Connaught Laboratories, Ltd.; CAV-1(CLL)] has been cloned in plasmid pAT153 in the form of subgenomic BamHI digestion fragments. Analysis of the nucleotide sequences of cloned terminal fragments has revealed an inverted terminal repeat (ITR) with a minimum length of 198 nucleotides, including a tandem reiteration of the 40-bp nucleotide sequence from positions 14 to 53. The ITRs had the 5'-CATCATCAAT ... sequence typical of adenoviruses and the highly conserved sequence ATAATATAC (nucleotides 9-17) of human strains. Additionally, one BamHI A clone (left terminus) contained three sequential copies of the 40-bp sequence, and two BamHI C clones (right terminus) contained at least seven. These did not appear to be artifacts of cloning, since evidence was obtained that the multiple reiterations also occurred in DNA isolated from intact virus. By analogy with human adenoviruses, the repetitive sequence in the CAV-1(CLL) genome encompasses the entire nuclear factor I (NFI) binding site of the origin of DNA replication. Additionally, the 40-bp nucleotide sequence was found to contain the sequence AGG(N)4GCCTAA (nucleotides 27-39), which closely resembles the concensus sequence of the human adenovirus NFI binding site [TGG(N)6-7GCCAA; nucleotides 25-38]. It appears, therefore, that the Connaught CAV-1 vaccine contains reiterated copies of an essential part of the adenoviral origin of DNA replication. A mechanism is proposed for the generation of multiple reiterations of sequences in the right ITR, given an initial single tandem repeat in the left ITR. PMID- 3037785 TI - Restriction enzyme accessibility and RNA polymerase localization on transcriptionally active SV40 minichromosomes isolated late in infection. AB - The transcriptionally active SV40 minichromosomes isolated late in infection contain a nucleosome-free ORI region or gap. To analyze the chromatin structure of this subpopulation of minichromosomes extracted at different ionic strengths in the early and late coding regions, minichromosomes were isolated in the presence of a 5, 50, or 130 mM concentration of monovalent cations and subjected to in vitro RNA elongation in either the presence or the absence of high salt and anionic detergent. The minichromosomes isolated at low ionic strength were transcriptionally more active than those isolated at physiological ionic strength. Nevertheless, in each case, the in vitro elongation complexes were present essentially on the late strand of the SV40 genome and localized preferentially in the late and 3' early coding regions. These regions were transcribed similarly in either the presence or the absence of chromatin denaturing agents. In contrast, the in vitro elongation activity of the RNA polymerase molecules present on the late strand in the middle and 5' end of the early coding region was inhibited in the absence of treatments to disrupt chromatin structure. In addition, as probed by restriction enzyme digestion, the ORI and late coding regions of the transcriptionally active minichromosomes were found to be more sensitive than the 5' region of the early genes. Taken together, these results suggest that the 5' and middle regions of the early genes of the SV40 transcriptional complexes isolated late in infection at low or physiological ionic strength are packaged in a more compact conformation than the rest of the genome. PMID- 3037786 TI - [The potential and significance of pharmacologic effects on leukotrienes and other lipoxygenase derivatives of arachidonic acid]. PMID- 3037787 TI - [Initial results of testing of an experimental sample of the Soviet kit for diagnosing viral hepatitis A]. PMID- 3037788 TI - [Significance of the level of specific antibodies in the development of the icteric and inapparent forms of viral hepatitis A]. PMID- 3037789 TI - [Effect of carbon monoxide on adenyl cylase and cyclAMP phosphodiesterase activity in the rat (experimental study)]. PMID- 3037790 TI - [The effect of gammaphos (WR-2721) on survival in combined radiation injuries]. PMID- 3037791 TI - [Effect of vasopressin on the Na,K-ATPase activity in synaptic membranes of the brain of adult and old rats]. AB - Vasopressin regulated in vitro the activity of Na+, K+-ATPase in synaptic membranes from rat brain cortex and brain stem. Low concentrations of the peptide activated the enzyme, high concentrations--inhibited. The effect was most distinct in brain stem. Age-dependent effects of vasopressin on the Na+, K+ ATPase activity were observed. Basal activity of the enzyme was not altered in synaptic membranes with ageing. PMID- 3037792 TI - [The use of pharmacological preparations for the study of calmodulin interaction with enzymes]. AB - Calmodulin-dependent regulation of cyclic nucleotide phosphodiesterase and kinase phosphorylase activities as well as Ca2+-dependent regulation of kinase phosphorylase, mediated via the integrated calmodulin, were studied in presence of phenothiazine and butyrophenone series of pharmacological drugs. As compared with butyrophenones, phenothiazines were shown to be more effective inhibitors of calmodulin-dependent activation of the phosphodiesterase. Phenothiazines inhibited similarly the effect of calmodulin on activity of kinase phosphorylase, whereas they did not affect the Ca2+-dependent activity of kinase phosphorylase. At the same time, butyrophenones proved to inhibit the Ca2+-dependent activation of kinase phosphorylase, mediated via integrated calmodulin as well as these drugs inhibited uniformly the calmodulin-dependent regulation of both kinase phosphorylase and phosphodiesterase. The data obtained suggest that dissimilar effect of phenothiazines on calmodulin-dependent regulation of kinase phosphorylase and phosphodiesterase, carried out using dissimilar mechanisms, required an extreme caution in evaluation of physiological and biochemical experiments, where these drugs were used as means for study of calmodulin functions in biological processes. PMID- 3037793 TI - [Metabolic pool of free amino acids in the blood and cerebrospinal fluid of patients with pituitary adenoma before and after proton therapy]. AB - A pool of free amino acids was studied in blood and cerebrospinal fluid of 34 patients with adenoma of hypophysis before proton therapy and within 3 years after the treatment. The course of proton therapy was shown to cause a regression of the disease, a decrease in content of somatotropic hormone in blood as well as to normalize the metabolic pool of amino acids in blood and cerebrospinal fluid. PMID- 3037794 TI - [Influenza virus C and its characteristics]. PMID- 3037795 TI - [Proteins of the human parainfluenza virus]. AB - Proteins of human parainfluenza type 3 virus propagated in green monkey kidney cultures are described. The presence of 4 major and 3 minor polypeptides is described along with their molecular weights. Protein F1 was found as a double band, its both components having a similar intensity. Only the upper band of F1 protein was found in the virus grown in Vero cell culture which has a low sensitivity for this virus. Fractionation of the virus and analysis of the proteins of subviral structures were carried out. Nucleocapsid had a buoyant density of 1.32 g/cm3 in cesium chloride and contained major NP protein and minor P and L proteins. PMID- 3037796 TI - [Diagnosis of human rotavirus infection by the dot hybridization method]. AB - The use of the dot-hybridization assay for the diagnosis of human rotavirus infection was shown to be principally possible. 32P-labeled RNA segments of porcine rotavirus transcribed in an in vitro system by activation of endogenous RNA-dependent RNA-polymerase were used as hybridization probes. The transcripts were synthesized to all 11 segments of the parental virus genome. The method proved to be highly specific and very sensitive detecting 10 picograms of viral RNA in fecal filtrates of the patients. PMID- 3037797 TI - [Persistent infection caused by the Coxsackie B3 virus in adult mice]. AB - A model of persistent infection with Coxsackie B-3 virus was developed in adult mice with clinical manifestations of the disease and long-term (up to 13 months) excretion of the causative agent. The method of multiple organ cultures was shown to be suitable for isolation of the persisting enterovirus. The presence of persistent infection was confirmed by the detection of IgM antibody in repeated daily examinations of the animals for 4 months. It seems to be expedient to use this model for investigations of the pathogenesis and methods of treatment of persistent Coxsackie B-3 infection in experimental animals. PMID- 3037798 TI - [Transforming activity of the DNA from human hepatoma cells. The transcription of integrated viral sequences and their cloning]. AB - A monolayer culture of mouse fibroblasts, line NIH/3T3, was transformed by DNA from human hepatoma cell line PLC/PRF/5. The resulting lines of transformed cells differ in their growth characteristics and are capable of producing tumors in nude mice. DNA of these cell lines was found to contain integrated sequences of human DNA and hepatitis B virus DNA. Analysis of transcription showed the amount of virus-specific RNA in the original cell line PLC/PRF/5 to be 20 times as large as in one of the transformed lines. Fragments of DNA of the transformed cells containing virus sequences were cloned, and 400,000 clones were obtained. A clone was identified the insert of which contains a portion of viral genome. PMID- 3037799 TI - [Detection of HTLV-III-specific sequences in T-lymphocyte lines by molecular hybridization]. AB - The methods of molecular hybridization are described using 32P-labeled plasmid pBHIO carrying the cloned HTLV-III provirus employed for the detection of HTLV III-specific sequences in lines of T-lymphocytes from patients with acquired immune deficiency syndrome. The presence of virus-specific sequences was demonstrated in 13 out of 20 lines examined by the dot-blot hybridization method. The blot-hybridization method revealed differences in sites of Hind III restriction in 2 out of 6 lines studied. The possibility of combined infection of line No. 17 with two different variants of HTLV-III is discussed. PMID- 3037800 TI - [Indices of the immune status of recurrent herpes patients being treated with a herpes polyvaccine]. AB - Patients with dermal and genital forms of recurring herpes were found to have decreased levels of leukocytes and changed balance of the peripheral blood lymphocyte populations. Treatment with herpes inactivated cultural polyvaccine produced a therapeutic effect in a significant number of patients which was accompanied by a correcting effect of the vaccine on their immune status. PMID- 3037801 TI - [Activation of a herpetic infection via corneal transplantation]. AB - Activation of herpes infection in keratoplasty was studied in 26 patients and found to be caused both by operative intervention and the corresponding corticosteroid therapy. The fluorescent antibody technique proved to be the most effective and rapid test for the detection of herpes infection. It is recommended to carry out laboratory control in transplantation of the cornea as well as topical antiviral therapy in the postoperation period not only in patients with herpetic keratitis but also in the subjects operated on for nonspecific leukoma and other diseases of the cornea. PMID- 3037802 TI - [Preliminary study results on the immunogenicity of a subunit herpes vaccine]. PMID- 3037803 TI - [Analysis of the electrophoretic types of human rotaviruses isolated in the city of Gorki]. PMID- 3037804 TI - [Articular manifestations in patients with Reiter's syndrome]. AB - The polyetiology, the discrepancies in the initial manifestations, polymorphism in the course and the possibility of the ephemerality or absence of ocular, uro genital and skin-mucosal manifestations create jointly the preconditions for a frequent diagnostic error in Reiter syndrome. The study confirmed the thesis that the diagnostic characteristics including the articular symptomatics are among the most striking ones. Synovial-arthritis and the involvement of the soft tissues are accompanied by disorders in the immune system with increased number of early T-lymphocytes, of Fc Gd and complement (C3b)-receptor lymphocytes and of spontaneously blast-transformed cells. The rheumatological process is characterized by proportionately accumulation of 99m Tc-pertechnetate and 99m Tc pyrophosphate in the affected articular structures, with asymmetry of affection, with predilection and maximal capture of the radiopharmaceutical in the feet. The radioisotope information is on scanty alterations in the spine and sacroiliac joints. The complex study facilitates the timely diagnosis of the disease. PMID- 3037805 TI - [Endocrinology of mastopathy and small breast cancer]. AB - The role of hormones in the cancerogenesis of breast cancer is still discussed. It is not clear at the moment if sexual hormones are primarily or secondarily in wall in cancerogenesis. Beside the discussion it is clear that estrogen, progesterone, androgens, prolactin and thyroxin have influences on the physiology and pathophysiology of the breast tissue. Furthermore the isoforms of these hormones and also the sexual hormone, binding-protein could be of interest. At last also the receptor evaluation becomes of interest in the etiology and therapy of breast cancer. PMID- 3037806 TI - Species specificity in the metabolism of nabilone. Relationship between toxicity and metabolic routes. AB - The disposition of 14C-nabilone has been examined in dogs and monkeys; it is rapidly absorbed and extensively metabolized in both species. A metabolic pathway initiated by the stereoselective enzymic reduction of the 9-keto moiety of nabilone was of major importance in the biotransformation of nabilone in the dog but was only a minor pathway in the monkey. The resulting long half-lived carbinol metabolites accounted for 77% of the circulating 14C in dog but for only 19% in monkey, 48 h after drug administration. Accumulation of carbinol metabolites in brain tissue was observed in dogs, with 24 h brain concentrations being five to six times higher than the plasma concentrations. No accumulation of carbinol metabolites, nabilone or of 14C occurred in the brain of monkeys. Accumulation of the carbinol metabolites in brain tissue has been implicated as the causative factor for the CNS toxicity observed in dogs treated chronically with nabilone. Comparison of nabilone pharmacokinetics in dog and humans showed the dog to be unique in producing high levels of carbinol metabolites, so that monkey appeared to be a more appropriate model than dog for pre-clinical toxicological and safety studies. Studies with liver 15,000 g supernatants indicated that the enzymic reduction of nabilone to its carbinol metabolite was a viable metabolic pathway in rat, dog and monkey, so that a distinct species difference exists between dog and monkey in the subsequent metabolism and/or elimination of these carbinol metabolites. PMID- 3037807 TI - [Surgical treatment of bronchial cancer--current status]. AB - It is still difficult to codify the surgical treatment of primary bronchogenic cancer. In spite of improvements of means of diagnosis and of detection of metastases the results at intermediate and long term are frequently modest. The techniques of resection are improved, and this is the method of choice in all cases where function and anatomical situation are permissive. The risk of the operation is minimal if the respiratory function is well, even with very extended resections. This is the greatest progress during the last 30 years. Surgery can cure only tumors which are still localized. Research on complementary treatment of extended forms is still necessary. PMID- 3037808 TI - [Neurologic diseases transmitted by ticks]. AB - Apart from causing various dermatological disorders, ticks, which also widespread in our part of the world, can transmit diseases that affect the nervous system. The difficulty of carrying out differential diagnosis and the possibility of administering causal treatment in cases of meningopolyneuritis Garin-Bujadoux Bannwarth and progressive Borrelian encephalomyelitis are giving rise to demands for an improvement in immunodiagnosis. PMID- 3037809 TI - E. coli maltodextrin phosphorylase: primary structure and deletion mapping of the C-terminal site. AB - The complete 796 residue amino acid sequence of maltodextrin phosphorylase was deduced from the E. coli malP nucleotide sequence. The calculated molecular weight of 90,500, including pyridoxal phosphate, is significantly larger than experimentally determined values. Enzymatically active and inactive mutants following deletion or exchange of up to 8 codons (7 amino acids) at the 3' end (C terminus) confirm the size of the mature native enzyme and disclose the essential functional or structural role of the highly conserved C-terminal region of phosphorylases. PMID- 3037810 TI - 6-Methylpurine, 6-methyl-9-beta-D-ribofuranosylpurine, and 6-hydroxymethyl-9-beta D-ribofuranosylpurine as antiviral metabolites of Collybia maculata (Basidiomycetes). AB - 6-Methylpurine, 6-methyl-9-beta-D-ribofuranosylpurine and 6-hydroxymethyl-9-beta D-ribofuranosylpurine were isolated from mycelial cultures of Collybia maculata and their antifungal, cytotoxic and antiviral activities investigated. This is the first report on the natural occurrence of these compounds. PMID- 3037811 TI - Pyrimidine homoribonucleosides: synthesis, solution conformation, and some biological properties. AB - Conversion of uridine and cytidine to their 5'-O-tosyl derivatives, followed by cyanation with tetraethylammonium cyanide, reduction and deamination, led to isolation of the hitherto unknown homouridine (1-(5'-deoxy-beta-D allofuranosyl)uracil) and homocytidine (1-(5'-deoxy-beta-D allofuranosyl)cytosine), analogues of uridine and cytidine in which the exocyclic 5'-CH2OH chain is extended by one carbon to CH2CH2OH. Homocytidine was also phosphorylated to its 6'-phosphate and 6'-pyrophosphate analogues. In addition, it was converted, via its 2,2'-anhydro derivative, to arahomocytidine, an analogue of the chemotherapeutically active araC. The structures of all the foregoing were established by various criteria, including 1H and 13C NMR spectroscopy, both of which were also applied to analyses of the solution conformations of the various compounds, particularly as regards the conformations of the exocyclic chains. The behaviour of the homo analogues was examined in several enzymatic systems. Homocytidine was a feeble substrate, without inhibitory properties, of E. coli cytidine deaminase. Homocytidine was an excellent substrate for wheat shoot nucleoside phosphotransferase; while homouridine was a good substrate for E. coli uridine phosphorylase. Although homoCMP was neither a substrate, nor an inhibitor, of snake venom 5' nucleotidase, homoCDP was a potent inhibitor of this enzyme (Ki approximately 6 microM). HomoCDP was not a substrate for M. luteus polynucleotide phosphorylase. None of the compounds exhibited significant activity vs herpes simplex virus type 1, or cytotoxic activity in several mammalian cell lines. PMID- 3037812 TI - Immunogenicity and safety of a low passage level bovine rotavirus candidate vaccine RIT 4256 in human adults and young infants. AB - A candidate rotavirus vaccine RIT 4256, derived from Nebraska calf diarrhea virus by 21 tissue culture passages, was tested in humans and compared with the RIT 4237 vaccine derived from the same stem virus by 147 tissue culture passages. The low passage strain RIT 4256 was first tested in adult volunteers for immunogenicity and safety: a serological response was seen in 9/18 (50%) vaccinees; three subjects had a mild fever reaction attributable to the vaccine. In 6 month old children the RIT 4256 vaccine elicited a serological response in 12 of the 21 (57%) seronegative recipients; two children had a possible fever reaction from the vaccination. In newborn infants a serological response following vaccination was detected in 19/41 (46%) of the recipients of the RIT 4256 and in 19/40 (48%) of the RIT 4237 vaccine; none of the newborn infants had any reaction from either vaccine. It is concluded that the low passage strain RIT 4256 is not more immunogenic than the high passage vaccine RIT 4237 in humans. The vaccines do not differ in clinical reactogenicity for man. PMID- 3037813 TI - Effect of current Japanese encephalitis vaccine on different strains of Japanese encephalitis virus. AB - Inactivated Japanese encephalitis (JE) vaccine was given to healthy children and their neutralizing antibody titres against nine JE virus strains were measured. After two shots of the vaccine in the first year, most of the children showed seroconversion, but their neutralizing antibody titres were not high, and one year after vaccination their titres had decreased to low levels. However, a booster dose of vaccine in the second year had a marked effect on the antibody response. The neutralizing antibodies induced by the vaccine reacted to almost the same extent with nine different JE virus strains, though significantly higher titres against the Nakayama strain, the vaccine strain, were observed after vaccination in the second year. PMID- 3037814 TI - Feline leukaemia vaccine protection against viral latency. AB - Seventeen cats, which were previously vaccinated with a subunit, feline leukaemia vaccine (Leukocell) and subsequently challenged with virulent feline leukaemia virus (FeLV), were tested at 2 to 4 years postchallenge for reactivation of latent FeLV infections. Administration of weekly doses of methylprednisolone induced significant decreases in lymphocyte numbers, but did not reactivate virus in bone marrow cultures from 15 cats in vivo or in vitro. These cats were observed to be neither persistently or latently viraemic prior to corticosteroid administration. The results of this study indicate that the vaccine is effective in affording significant protection against latent FeLV infections, even after severe immunosuppression. This finding, coupled with previously published results indicating protection against persistent viraemia and tumour formation, makes this vaccine highly effective in protecting against FeLV infections and associated disease. PMID- 3037815 TI - Intestinal immunity induced by inactivated poliovirus vaccine. AB - In order to detect any intestinal immunity against poliovirus infection induced by parenteral vaccination with inactivated poliovirus vaccine (IPV), experiments were conducted in monkeys (Macaca radiata). All animals were seronegative (antibody not detected at 1:2 dilution of serum) before investigation. Sixteen monkeys were vaccinated with three doses of IPV at monthly intervals. Groups of four vaccinated and two seronegative control animals were fed 100 median monkey infectious doses (100 MID50) of poliovirus type 1 (Mahoney strain) at 1, 4, 7 and 12 months after vaccination. While all control monkeys excreted poliovirus in the throat and faeces from day 2 to days 18-22, none of the vaccinated monkeys excreted virus. Thus, a high degree of intestinal immunity against infection was found. Although no evidence of infection was seen, antibody booster response occurred in most monkey fed virus 7 and 12 months after vaccination. PMID- 3037816 TI - [Problems of pathogenesis and treatment of vitiligo]. PMID- 3037817 TI - [Role of ACTH, somatotropic hormone, cortisol and insulin in the pathogenesis of lupus erythematosus]. PMID- 3037818 TI - Nucleotide sequence of the gene encoding the Newcastle disease virus hemagglutinin-neuraminidase protein and comparisons of paramyxovirus hemagglutinin-neuraminidase protein sequences. AB - The nucleotide sequence of cloned cDNA copies of the mRNA encoding the Newcastle disease virus (NDV), strain A-V, hemagglutinin-neuraminidase (HN) protein was determined. A single open reading frame in the sequence encodes a protein of 570 amino acids with a calculated molecular weight of 62,280. The predicted protein sequence contains only one obvious potential membrane spanning region, located 27 amino acids from the amino terminus of the sequence. The predicted sequence contains 6 glycosylation sites and 14 cysteine residues. Comparison of the NDV HN protein sequence with three other paramyxovirus HN protein sequences reveals two regions that have homologies in all four sequences. The conserved cysteine residues are clustered in these two regions. One conserved region is located near the middle of the predicted sequence while the second region is in the carboxy terminal third of the molecule. The presence of conserved regions suggests the importance of these areas of the molecule in the structure or function of the protein. PMID- 3037819 TI - Response of genetically susceptible and resistant mice to intranasal inoculation with mouse hepatitis virus JHM. AB - Mouse hepatitis virus (MHV)-JHM infection was studied in genetically susceptible (BALB/cByJ) and resistant (SJL/J) mice following intranasal inoculation at 1, 3, 6 or 12 wk of age. Markers of infection included histology, immunohistochemistry, virus quantification and virus serology. All BALB mice developed severe disseminated disease with high mortality due to encephalitis and hepatitis. Peak MHV titers appeared in brain, liver, spleen and intestine on days 3 or 5. Age at inoculation did not influence virus titers in brain, spleen or intestine, but virus titers in liver were inversely proportional to age at inoculation. In 6-wk old BALB mice, virus was cleared from spleen, intestine and liver by day 30 and from brain by day 60. In intestine, MHV was localized to lymphoid tissue, without fecal excretion. SJL mice of all ages developed remarkably milder disease with low mortality occurring only among mice inoculated at 1 wk of age. SJL mice inoculated at 1 wk had disseminated infection at day 3, but lesions and antigen were cleared from most organs by day 5. Mice inoculated at 3 and 6 wk of age had minimal or no involvement of peripheral organs, and mice inoculated at 12 wk of age had infections restricted to the nose. At day 5, MHV titers in brain, liver, spleen and intestine were significantly lower or undetectable in SJL mice of all ages compared to age-matched BALB mice. In 6-wk-old mice, MHV was cleared from all organs by day 10. Serum antibody titers to MHV were many-fold higher in BALB mice, compared to SJL mice, which mounted only a modest response. PMID- 3037820 TI - Effect of lysosomotropic compounds on early events in foot-and-mouth disease virus replication. AB - The effect of three lysosomotropic compounds, chloroquine, monensin and NH4Cl, on the replication of foot-and-mouth disease virus (FMDV) type A12 was studied. Viral replication was almost totally inhibited by 0.5 mM chloroquine, 50 microM monensin, or 25 mM NH4Cl. Monensin and NH4Cl affected replication when added either before or within the first hour of infection. Chloroquine, however, still inhibited viral replication when added up to 2.5 h after infection. Assays of binding of radiolabeled virus to cells showed that these compounds had no effect on viral adsorption. Neither monensin nor NH4Cl had any significant effect on cellular protein synthesis, but there was no evidence of viral protein synthesis in cells infected in the presence of these compounds. In contrast, chloroquine inhibited both cellular and viral protein synthesis. Eclipse assays, performed in the presence of the compounds, showed that while chloroquine and NH4Cl had little effect on cell-induced degradation of incoming virions to 12 S protein subunits, monensin inhibited this reaction. The replication of representative members of all seven serotypes of FMDV was inhibited by monensin although some types were less sensitive to the compound than others. These results are consistent with a model which postulates that viral eclipse is the result of acidification of endocytic vesicles which degrade entrapped virions to 12 S protein subunits resulting in the release of genome RNA. PMID- 3037821 TI - Cold-adaptation of human rotavirus. AB - A human rotavirus strain was cold-adapted for possible future use as a live vaccine. The original strain was isolated in 1980 in primary cynomolgus monkey kidney cells and has a serotype I and subgroup II antigenicity. The virus was serially passaged in African green monkey kidney cells; it was cultivated at 37 degrees C at the first stage of passages, and the cultivation temperature was then shifted down stepwise by 3 degrees C per each 10 passages. Finally the virus was passaged 10 times at 25 degrees C (total passage number of 55). The virus formed small-size plaques with irregular shaped borders at 31 degrees C. Growth at 25 degrees C of the cold-adapted virus was higher than that of the original virus. There was no difference between the migration patterns of 11 dsRNA segments in polyacrylamide gel electrophoresis of the original and the cold adapted viruses. PMID- 3037822 TI - Plasma protein and red cell enzyme groups in Galicia (north west Spain). AB - Galactose-1-phosphate uridyl transferase (GALT), esterase D (EsD), and plasminogen (PLG) phenotypes were determined by isoelectric focusing in thin layer polyacrylamide gels (PAGIF) in a random sample from Galicia. Haptoglobins (Hp) were determined by conventional electrophoresis. The following gene frequencies were observed: for GALT: GALTN: 0.930; GALTD1: 0.044; GALTD2: 0.025; for EsD: EsD1: 0.874; EsD2: 0.104; EsD3: 0.021; for PLG: PLG1: 0.800; PLG2: 0.199; for Hp: Hp1: 0.426; Hp2: 0.573. Population data results of all electrophoretic markers typed until now in Galician population are also included. PMID- 3037823 TI - [Circadian rhythm of cortisol and corticotropin (ACTH) in patients with rheumatoid arthritis in relation to inflammatory activity]. AB - In patients with rheumatoid arthritis, corticosteroids are usually given as a single morning dose to minimize adrenal suppression, because of the well-known circadian rhythm of the secretion of cortisol and corticotropin. However, it has not been so far examined whether there really is a normal secretion pattern of those hormones in all patients with rheumatoid arthritis, as there is in healthy subjects. We therefore studied twelve patients with rheumatoid arthritis who had never been treated before with corticosteroids and disease-modifying drugs such as gold, D-penicillamine or immunosuppressive drugs. The study was designed to examine adrenal secretory activity by measuring the cortisol and corticotropin levels at 2 h intervals throughout a 24 h cycle. The circadian secretion patterns of cortisol and corticotropin were disturbed in most patients. High inflammatory activity of the disease was accompanied by a severe disturbance of the circadian rhythm, whereas patients with low inflammatory activity showed a nearly normal secretion pattern. Besides other causes, the influence of mediators of inflammation on hypothalamic centers, analogous to endogenous pyrogen in fever, must be discussed. PMID- 3037824 TI - [Hydroxylapatite ceramic-granulate and its systematics]. PMID- 3037825 TI - [Experimental diabetic neuropathy. Morphometric studies of spinal ganglia cells in short-term streptozotocin-induced diabetes]. AB - Ganglion cells of both the left and right dorsal root ganglia L5 were investigated morphometrically in 8 diabetic rats 35 or 44 d after the administration of streptozotocin, as well as in 8 controls. The quantitative study included the analysis of mean neuron volume, maximal cell diameter, cell number per volume, by cell size class and the analysis of histograms. There were no changes in cell size. Whereas in experimental short-term diabetes the peripheral nerves demonstrate morphologic changes correlating to the biochemical abnormalities (reduced amino-acid uptake, decreased activity of Na+, K+-ATPase) no such correlations can be observed in the perikarya. PMID- 3037826 TI - [Concurrent appearance of stomach polyposis and hepatocellular carcinoma]. AB - An 18 year old woman had been known to have juvenile polyposis since five years with multiple hyperplastic polyps of the stomach and several polyps of the colon. Death occurred from hepatic coma with hepatocellular carcinoma. A lymphangiosis carcinomatosa was observed in some stomach polyps with a circumscribed infiltration into the stroma and into the adjacent glands. Such an unusual combination of diseases readily causes diagnostic confusion. PMID- 3037827 TI - The efficiency of various laboratory methods for the diagnosis of adenovirus conjunctivitis. AB - Virological and serological data from 242 patients with conjunctivitis or keratoconjunctivitis, infected with adenovirus type 8, and of 109 patients infected with other adenovirus types were compared by means of an electronic data processing system. About 55% of the patients in both groups showed a diagnostic antibody rise in genus-specific (antihexon) complement-fixation. This rate is significantly lower than that obtained with genus-specific ELISA IgG or IgA, or with neutralization tests against the homologous virus, or with hemagglutination inhibition in case of adenovirus 8 infection. All these tests showed titer rises in 72 to 85% of the patients; differences were not significant. Virus isolation from conjunctival swabs, taken up to 9 days after onset of disease, was successful in 80 to 90%, thereafter in 30%. The time-course of the various antibody titers showed that genus-specific (antihexon) antibodies mostly rise 9 to 12 days after onset, whereas type-specific neutralizing or hemagglutination inhibiting antibodies rise 10 to 14 days after onset. PMID- 3037828 TI - Similarities between soluble inorganic pyrophosphatase from yeast and some nucleotide-binding polypeptides. PMID- 3037829 TI - Detection of human papillomavirus in cervical smears. A comparison of in situ hybridization, immunocytochemistry and cytopathology. AB - A comparison of different methods for the detection of cervical human papillomavirus (HPV) infection was made on patients attending the cervical dysplasia clinic. Cytomorphology, immunocytochemistry and in situ hybridization were compared for their ability to detect HPV. Separate cervicovaginal smears from 50 patients were tested for HPV types 6/11, 16 and 18 by in situ hybridization using 35S-labeled DNA probes. Duplicate smears from the same patients were Papanicolaou stained and evaluated for evidence of condylomatous and dysplastic changes. Twenty-five matching cervical biopsies were immunostained for HPV capsid antigen and tested by in situ hybridization for HPV DNA. The cytologic smears of 20 patients (40%) were positive for HPV DNA. Six patients had HPV 6/11, ten had HPV 16, three had HPV 18, and one had both HPV 6/11 and HPV 16. There was a high correlation between condylomatous cytopathology and antigen and DNA detection. One-third of the specimens with condylomatous changes were DNA negative by the tested probes, suggesting the presence of other HPV types in the genital tract. PMID- 3037830 TI - A morphologic analysis of the cells of ductal carcinoma of the breast and of adenocarcinoma of the ovary in pleural and abdominal effusions. AB - The cells in serous effusions from 14 cases of histologically proven ductal adenocarcinoma of the breast and from an equal number of proven adenocarcinomas of the ovary were morphologically analyzed, with quantitative values assigned to various cell parameters. The resulting data were then subjected to stepwise discriminant analysis. The presence of coarse chromatin proved to be the most significant discriminating factor, resulting in a correct classification of 100% of the mammary carcinomas (14 of 14) and 71.4% (10 of 14) of the ovarian carcinomas, for an overall rate of 85.7% correct classifications. When combined with other cell parameters, such as the presence of polygonal cells, multiple nucleoli and an isolated cell arrangement, the accuracy rate increased to 92.9% correct classifications. Ovarian carcinoma was more likely to have coarse chromatin and to occur in three-dimensional cell groups, in contrast to mammary carcinoma, which tended to have fine chromatin and more polygonal cell shapes and to occur more commonly in syncytial cell groups and as isolated cells. PMID- 3037831 TI - The cytodiagnosis of well-differentiated neuroendocrine carcinoma. A distinct clinicopathologic entity. AB - Well-differentiated neuroendocrine carcinoma (WDNE) has been recognized as a distinct variant of pulmonary neuroendocrine carcinoma, with characteristic histopathologic and clinical features that separate it from both carcinoid and small-cell carcinoma (SCC). Histologic review of tumors in long-term survivors (greater than two years) with an initial diagnosis of SCC has shown that the majority of these cases are, in fact, better classified as WDNE; the distinction of WDNE from SCC has, therefore, important prognostic and therapeutic implications. A retrospective review of 200 cytologies originally diagnosed as SCC was undertaken in an attempt to characterize the cytomorphologic features of WDNE. The cytologic criteria that distinguished cases of WDNE included polygonal to-fusiform cells with a variable amount of lacy cytoplasm, oval nuclei with coarsely dispersed chromatin and frequent chromocenters, and mild nuclear and cytoplasmic anisomorphism. The majority of malignant cells were arranged either in acinarlike clusters or in epithelial sheets with evidence of palisading. Twenty-two cases were reclassified cytologically as WDNE and were accurately distinguished from all other neoplastic and SCC cases on repeated double-blind review. Clinical and histologic data confirmed the diagnosis of WDNE in all cases; it can be concluded that SCC and SDNE are cytologically distinct entities. PMID- 3037832 TI - Fine needle aspiration cytology of atypical carcinoid of the lung. AB - The cytologic features of eight atypical carcinoid tumors of the lung, as observed in fine needle aspiration (FNA) specimens, are described in detail. They were compared with 21 pulmonary squamous-cell carcinomas, 16 adenocarcinomas, 5 small-cell undifferentiated carcinomas, 3 large-cell undifferentiated carcinomas and 1 typical carcinoid tumor. Atypical carcinoid tumor was easily distinguished from the other pulmonary neoplasms in most instances. Only two poorly differentiated squamous-cell carcinomas (one of which had atypical carcinoid as a component) and one small-cell undifferentiated carcinoma had similar cytologic features. One atypical carcinoid also had cytologic features similar to small cell undifferentiated carcinoma. Because atypical carcinoid and small-cell undifferentiated carcinoma, at times, may be difficult to separate in FNA specimens, surgical resection of all stage I neoplasms with cytologic features evocative of either neoplasm is recommended. PMID- 3037833 TI - Cytologic features of islet-cell tumors. AB - Although a number of reports have demonstrated the accuracy of fine needle aspiration (FNA) in the diagnosis of nonendocrine pancreatic carcinomas, the cytomorphology of islet-cell tumors (pancreatic endocrine tumors) is not well defined. This paper describes the cytologic features of three histologically confirmed cases of islet-cell tumors. The three tumors occurred in one man and two women, who were 63, 64 and 70 years of age, respectively. Each patient underwent FNA of pancreatic mass with computed body tomography guidance. The aspirates contained large numbers of tumor cells in two cases and a smaller number in one case. The cells, distributed singly and in small groups, were small and round or polygonal, with scant to more often abundant, dense or granular cytoplasm. The nuclei were often located eccentrically and were round to oval, with smooth nuclear borders and finely stippled chromatin. Many nuclei contained a single nucleolus. Multinucleated cells were present and generally contained two or three nuclei. Although the diagnosis of islet-cell tumors in fine needle aspirates is difficult, the cytomorphologic features of these tumors are sufficiently distinctive to suggest the diagnosis, especially when a relatively monomorphic population composed predominantly of single cells is present. PMID- 3037834 TI - Physiological aspects of the thyroid trapping function and its suppression in iodine deficiency using 99mTc pertechnetate. AB - In an area of iodine deficiency, we investigated 190 individuals with and without euthyroid endemic goitre, who had a normal TSH response after TRH and an entirely homogeneous thyroid scintigram before and under suppression. In these subjects, the thyroid uptake of 99mTc pertechnetate, as a measure of the iodide trapping function, was determined before (TcUb) and under suppression (TcUs), using quantitatively evaluated scintigraphy. In this control group of individuals, without evidence of autonomy, the reference ranges of TcUb and TcUs were determined. The upper limit of the reference range for TcUb was 7.4% of the tracer activity injected, and for TcUs 1.6%. The reference range of TcUs is to be used to detect accurately thyroid autonomy in vivo. In addition, factors affecting the thyroid trapping function were investigated. With decreased iodine supply, trapping before suppression was increased, compensating for iodine deficiency. The effect of TSH on the trapping function was secondary, indicating that, to a considerable degree, the follicular cells adapt their iodide trapping to the iodine supply. The trapping before and under suppression increased with the estimated thyroid weight. delta TSH after TRH stimulation measured before suppression correlated inversely with the trapping under suppression. The two latter observations suggest that there is a continuum between low and increased levels of the TSH-independent thyroid function, even among these selected individuals without primary evidence of autonomous tissue. A correlation of the trapping function with sex, oestrogen treatment, and goitre type was not demonstrable. Age was found to have a small influence. Except for iodine contamination, the factors affecting the pertechnetate uptake of the thyroid, can be neglected under routine conditions. PMID- 3037835 TI - Effects of high-dose ketoconazole and dexamethasone on ACTH-stimulated adrenal steroidogenesis in orchiectomized prostatic cancer patients. AB - The effects of high-dose ketoconazole (i.e. 400 mg every 8 h) therapy on adrenal steroidogenesis were investigated in 7 patients with advanced prostatic cancer who no longer responded to orchiectomy. An ACTH challenge was performed before and on days 14 and 28 of high-dose ketoconazole treatment. During the last 14 days, dexamethasone (0.5 mg twice daily) was administered together with ketoconazole. High-dose ketoconazole alone lowered the basal levels of the androgens by 49-66%. It almost completely inhibited their stimulation by ACTH, whereas plasma progesterone was doubled. Basal cortisol was only slightly lowered, but the response to ACTH stimulation was markedly blunted. Basal and stimulated plasma aldosterone remained unaffected. Both basal and stimulated 11 deoxycortisol, 11-deoxycorticosterone, and, to a lesser extent, corticosterone rose more markedly after ketoconazole than after placebo. The basal and stimulated plasma adrenal androgen levels were further reduced after combined ketoconazole-dexamethasone treatment, whereas plasma corticosterone, 11 deoxycortisol, and 11-deoxycorticosterone were lowered in the same way as cortisol. Aldosterone and progesterone profiles were similar to those observed under high-dose ketoconazole, but plasma 17 alpha-hydroxyprogesterone increased more markedly than after high-dose ketoconazole alone. These results demonstrate that high-dose ketoconazole lowers plasma androgen levels in orchiectomized patients and partly inhibits the gluco- and mineralocorticoid syntheses, especially after ACTH-stimulation. The addition of dexamethasone does not only correct the possible consequence of the impairment of the cortisol production by high-dose ketoconazole, but it further reduces the androgen levels and lowers the plasma concentrations of most precursors, for instance 11-deoxycorticosterone, which has some physiological mineralocorticoid activity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3037836 TI - Auer rod-like inclusions in prolymphocytic leukemia. AB - Auer rod-like inclusions (ARLI) can be seen in B cell malignancies but are very rare in prolymphocytic leukemia (PLL). Prolymphocytes with ARLI from a patient with PLL were examined using cytochemical and immunochemical techniques and transmission electronmicroscopy (EM). The lymphoblastic cells gave a positive reaction with antibodies to the gamma heavy chains and kappa light chain. On EM the ARLI were 1-2 micron long, consisting of election-dense crystalline matrix, with a periodicity of 60 A and contained within a single unit membrane. The production of IgG by PLL cells with ARLI has not been previously reported. It is emphasized that in PLL the lymphoblasts can have inclusions resembling Auer rods. Cytochemical and immunochemical studies should be done to avoid a misdiagnosis of acute nonlymphoblastic leukemia. PMID- 3037837 TI - Diminished release of lactoferrin from polymorphonuclear leukocytes of human neonates. AB - In neonatal and adult polymorphonuclear leukocytes (PMN) we determined the content and the release of beta-glucuronidase, myeloperoxidase, lysozyme and lactoferrin. We found an equal total content of these proteins in adult and neonatal PMN, except for a lower lysozyme concentration in neonatal PMN. In the presence of opsonized zymosan myeloperoxidase, lysozyme and beta-glucuronidase were released in equal amounts; lactoferrin, however, was released to a lower rate from neonatal than from adult PMN (p less than 0.0005). PMID- 3037838 TI - Quantitative cytochemical analysis of cytochrome oxidase and succinate dehydrogenase activity in spinal neurons. AB - This paper describes cytophotometric determinations of cytochrome oxidase (COX) and succinate dehydrogenase (SDH) activities in neurons in 3 areas of the spinal motor column of the teleost fish Brachydanio rerio (the Zebrafish). Purpose of this investigation was to analyse the correlation between the oxidative metabolic capacity of motoneurons with their activity patterns. The spatial organization of the spinal cord of the zebrafish allows such an analysis, because the motoneurons which innervate different muscle fiber types (slow tonic red and fast phasic white, respectively) occupy separate areas of the motor column (Van Raamsdonk et al. 1983). We analysed the COX and SDH activities on serially sectioned neurons, We found large variations in the ratio of COX/SDH activity: the ratio was high for large neurons (in the "white" area) and low for small neurons (in the "red" area). These findings were contrary our expectations, because COX as well as SDH activity have been proposed as indicators for neuronal activity if both activities are reliable indicators, then their ratio should be constant. In addition, COX and SDH activities were analysed on serially sectioned anterior horn neurons of the cat spinal cord. In contrast to the situation in fish, we observed a statistically significant proportionality between COX and SDH activities. We conclude that the histochemical reactions for COX or SDH activity have no general validity as markers for the same type of neuronal activity. PMID- 3037839 TI - Ultrastructural tissue morphometry of the distribution of extracellular matrix vesicles in remodeling rat tibial bone six days after injury. AB - A study of the distribution of extracellular matrix vesicles on the 6th day of bone healing was performed by methods of transmission electron microscopy combined with computerized morphometry. The vesicles were detected on the electron micrographs and grouped according to their diameters, distance from the calcified front and type. The different types were selected as follows: vesicles with electron-lucent contents, i.e., 'empty'; vesicles with amorphous electron opaque contents, i.e., 'amorphous'; vesicles containing crystalline depositions, i.e., 'crystal', and vesicles containing crystalline structures with ruptured membranes, i.e., 'rupture'. Most of the vesicles were concentrated between diameters of 0.02 and 0.22 microns. Most of the vesicles were found within a distance of less than 3 micron from the calcified front. The vesicles were distributed according to their types: 'empty', 'amorphous', 'crystal' and 'rupture' in 14, 39, 34 and 13%, respectively. The diameters of the 'crystal' and 'rupture' vesicles were significantly larger than those of the 'empty' and 'amorphous' types. The sequence of distances from the calcified front was recorded as follows: 'rupture', 'crystal', 'amorphous' and 'empty', the 'rupture' type being the closest to the front. The results of the present study confirm the accepted hypothesis on calcification via extracellular matrix vesicles. It is thought that the cell secretes 'empty' vesicles that accumulate amorphous Ca and P to form a hydroxy-apatite crystal. This is followed by rupture of the vesicular membrane. The propagation of the process is accompanied by increase in the vesicular diameter and its approximation to the calcifying front. PMID- 3037840 TI - Relationship of adjuvants and swine influenza vaccine to experimental neuropathy in rabbits. AB - Experimental neuropathy, characterized by endoneurial edema and demyelination, was induced by inoculating rabbits with a combination of Freund's complete adjuvant (FCA), gangliosides, lecithin and cholesterol. A less severe demyelinating neuropathy could be induced by treatment with FCA alone but no significant change could be elicited by injection of swine influenza vaccine (SFV) alone. When FCA was combined with gangliosides, lecithins, cholesterol and SFV, neuropathy occurred, but the changes were less severe than if these agents were used without SFV. Sera were tested for myelin basic protein (MBP) and galactocerebroside (GC) antibodies in each experimental group. Neither SFV alone nor SFV combined with Freund's complete adjuvant, gangliosides, cholesterol and lecithin evoked significant antibody titers to MBP or GC. However, rabbits inoculated with FCA, gangliosides, lecithin and cholesterol had rising titers of antibody to both MBP and GC over the 3-month experimental period. One rabbit inoculated with FCA alone had significant antibody to MBP. The findings suggest that Freund's complete adjuvant alone can induce demyelination in the peripheral nerves of rabbits and that SFV may modulate the immune response acting either as an adjuvant or suppressant in the experimental demyelinating disease. PMID- 3037842 TI - Glycogen accumulations in the cerebral cortex in Alzheimer's disease. AB - The fine structure of granular glycogen bodies (GGB) within the grey matter of the temporal cortex of 11 patients with Alzheimer's disease is described. GGB measure up to 50 microns in diameter and consist of densely packed alpha or beta glycogen granules (never both), neither of which are membrane bound. They were noted in axons, both myelinated and unmyelinated (sometimes close to the dystrophic neurites of senile plaques), and also in other processes of indeterminate origin. Their appearance may relate to disturbances of axonal transport resulting from damage to terminals within evolving senile plaques. PMID- 3037841 TI - Leucoencephalopathy with multinucleated giant cells containing human immune deficiency virus-like particles and multiple opportunistic cerebral infections in one patient with AIDS. AB - A 29-year-old homosexual male with AIDS presented with progressive encephalopathy and cytomegalovirus (CMV) pneumonia. Neuropathological examination revealed toxoplasma abscesses in corpus callosum, basal ganglia and cerebellar white matter; demyelinating foci in the parietal white matter, with microscopic changes typical of progressive multifocal leucoencephalopathy and intranuclear papovavirus inclusions in oligodendrocytes; and lesions of subacute encephalitis in the periventricular regions with large cells positive by immunostaining for CMV. Diffuse myelin loss was observed in the cerebral white matter. Multinucleated giant cells were numerous in the demyelinated areas, they were also observed in close relationship with papova, CMV and Toxoplasma lesions. Immunostaining of these cells was positive for histiocyte markers and negative with the leucocyte common antigen monoclonal antibody. Some of them contained virus-like particles measuring around 100 nm similar to human immune deficiency virus (HIV) as observed in human brain. PMID- 3037843 TI - Fingerprint inclusions in normal fetal muscle. AB - In muscle disease, fingerprint inclusions are variously described as a diagnostic or non-specific alteration. Fingerprint inclusions identified in human fetal muscle suggest these bodies may be transient structures of developmental significance accounting for their infrequent occurrence in myopathies of diverse pathogenesis and clinical expression. PMID- 3037844 TI - Almitrine neuropathy. A nerve biopsy study of 8 cases. AB - We report the quantitative and qualitative, light and electron microscopic studies, including teased fiber preparations of nerve biopsies obtained from eight patients, treated with almitrine, presenting with the characteristic association of a sensory peripheral neuropathy with a recent body weight loss. The data were consistent with an axonal damage affecting myelinated fibers, predominantly large ones and to a lesser degree unmyelinated fibers, some degree of segmental demyelination. Marked axonal regeneration was observed when the nerve biopsy was delayed after withdrawal of the drug, Micro-angiopathy secondary to the formation of concentric lamellae from the basement membrane was observed in five patients suffering from chronic hypoxemia. Almitrine is an agonist of the chemoreceptors. The pathogenesis of the toxic neuropathy induced by this compound remains obscure. Clinical features do not fit with a purely hypoxic mechanism at the origin of the neuropathy. PMID- 3037845 TI - Effects of ionizing radiation on retinoblastoma and on the normal ocular fundus in infants. A photographic and fluorescein angiographic study. PMID- 3037846 TI - Tissue copper content in primary and metastatic liver cancers. AB - Tissue copper contents in 38 primary and 45 metastatic hepatic malignancies and 15 control livers were analyzed by atomic absorption spectrophotometry. The average copper content of 15 control livers was 23.1 +/- 13.0 micrograms/g dry weight (microgram/gdw). The copper content of five cholangiocellular carcinomas (CCCs) and 45 metastatic cancers was almost equal to the control level. Thirty three hepatocellular carcinomas (HCCs) contained a larger amount of copper (61.5 +/- 76.8 micrograms/gdw) than the control livers (p less than 0.05), but the copper content of HCCs showed a considerably wide variation. The average copper content of nine minute HCCs (126 +/- 112 micrograms/gdw) was significantly (p less than 0.05) higher than that of 24 large HCCs (37.2 +/- 39.9 micrograms/gdw). Histologically, orcein and paramethylaminobenzylidene rhodamine positive granules were seen in eight and four of nine minute HCCs, respectively. These granules were also found in some large HCCs, but were never found in CCCs and metastatic cancers. It was concluded that these excessive accumulations of copper and copper binding proteins might present a helpful finding to distinguish some cases of HCC, especially small HCC, from CCCs, metastatic cancers and hypertrophic regenerative nodules of cirrhotic livers. The significance and possible pathogenesis of these copper accumulations in HCCs require further studies. PMID- 3037847 TI - Lipid storage disease: Part III. Ultrastructural evaluation of cultured fibroblasts in sphingolipidoses. AB - For the purpose of evaluating electron microscopy of tissue culture in making the diagnosis of sphingolipidoses, an ultrastructural study was made on the cultured fibroblasts from 23 patients with the disorders. The characteristic cytoplasmic inclusions were observed in the cultured cells of Fabry disease, Tay-Sachs disease, Sandhoff disease, generalized gangliosidosis, Niemann-Pick disease, metachromatic leukodystrophy, and multiple sulfatase deficiency, and differ in fine structure with these diseases. All these cytoplasmic inclusions were surrounded by a single limiting membrane and enzyme cytochemically showed acid phosphatase activity, indicating their lysosomal origin. Ultrastructurally, the cytoplasmic inclusions showed pleomorphic osmiophilic inclusions in Fabry disease, membranous cytoplasmic bodies (MCB) in Tay-Sachs disease and Sandhoff disease, MCB and vacuolar inclusions containing finely reticulogranular materials in generalized gangliosidosis, myelin-like inclusions in Niemann-Pick disease, concentric lamellar inclusions in metachromatic leukodystrophy, and polymorphic cytoplasmic inclusions in multiple sulfatase deficiency. In the heterozygous carriers of Fabry disease, pleomorphic osmiophilic inclusions were also detected. However, any specific inclusions were not detectable in the cultured fibroblasts of Gaucher disease and Krabbe disease. Availability of electron microscopy in the cultured fibroblasts of sphingolipidoses is discussed. PMID- 3037848 TI - Salivary glands in long-term alloxan-diabetic rats. A quantitative light and electron-microscopic study. AB - Fifty untreated diabetic animals were compared with 58 age-matched non-diabetic controls. Reduced salivary gland weight was evident after one month's diabetes and this was unchanged after 12 months of diabetes. Submandibular/sublingual gland weight was proportional to the reduced body weight in the diabetic rats. Parotid gland weight, however, was proportionally more reduced. Only diabetic rats had lipid inclusions in the acinar cells of their submandibular glands and the morphometrically estimated amount of inclusions was positively correlated to the blood glucose level. Acinar cell size was significantly increased in long term diabetic rats as compared with short-term diabetic rats and controls. Capillary basement membrane width was significantly increased in long-term diabetic rats compared with age-matched controls and with short-term diabetic rats. Thus, both the degree and duration of diabetes have a major effect on salivary gland morphology in alloxan diabetic rats. PMID- 3037849 TI - Human monocyte and polymorphonuclear leukocyte chemotactic and chemokinetic responses to leukotriene B4 and FMLP. AB - Comparable investigations of the chemotactic and chemokinetic responses of purified monocytes (MO) and polymorphonuclear leukocytes (PMN) to leukotriene B4 (LTB4) and N-formyl-methionyl-leucyl-phenylalanine (FMLP) were made in this study. Using a sensitive and objective 51Cr-chemotactic assay, it was shown that both MO and PMN showed a bell-shaped response to LTB4 and FMLP, with a maximum response at 10(-8)M for both drugs. For PMN, the maximal response elicited by LTB4 was similar in magnitude to that produced by FMLP, whereas the MO chemotaxis induced by 10(-8)M FMLP was significantly higher than the response evoked by 10( 8)M LTB4. For both cell types, LTB4 at low concentrations (less than 10(-9)M) gave rise to higher chemotactic responses than FMLP. Chemokinesis was differentiated from chemotaxis, using a checkerboard system. At concentrations less than 10(-9)M the LTB4-evoked contribution of chemokinesis to the total migrational response was significantly higher than the chemokinetic contribution of FMLP. Preincubation with LTB4 produced only homologous chemotactic deactivation to subsequent LTB4 stimulation, whereas preincubation with FMLP resulted in diminished secondary response to both FMLP and LTB4. The degree of deactivation was dependent upon the dose of attractant used, with a LTB4 concentration of 10(-7)M leading to about 40% and 25% deactivation of PMN and MO, respectively. Preincubation with 10(-7)M FMLP led to about 50% and 32% suppression of the subsequent chemotactic response of PMN and MO, respectively. PMID- 3037850 TI - Vaccination-induced activation of human blood T cells suppressing pneumococcal polysaccharide-specific B cells. AB - In order to study the regulation of the human B cell response to pneumococcal polysaccharides (PPS), anti-PPS-secreting cells (SC) and Ig-SC were quantitated in unstimulated and EBV-stimulated cultures of blood mononuclear cells (MNC) established before and one and two weeks after vaccination. In unstimulated cultures established one week after vaccination, significant numbers of anti-PPS SC were detected; however, they were observed only in cultures depleted of T cells (mean: 62/10(6)), suggesting the presence of T cells suppressing in vivo activated PPS-specific B cells. In EBV-stimulated cultures depleted of T cells, low numbers of anti-PPS-SC were observed before the vaccination, and the numbers of these cells increased significantly two weeks after the immunization (from 28 to 48/10(6)). In contrast, the numbers of anti-PPS-SC and total Ig-SC decreased in EBV-stimulated cultures of unseparated MNC established after the vaccination, suggesting T cell-mediated suppression. The findings were not explained by changes in the numbers of blood T helper (Leu 3+) or T suppressor (Leu 2+) cells following the vaccination. Possibly the vaccination induces the appearance in the blood of activated suppressor cells responsible for the suppression of B cells in unstimulated and EBV-stimulated cultures. PMID- 3037851 TI - [Biochemical study of Chinese rhubarb. XIX. Localization of inhibition of anthraquinone derivatives on the mitochondrial respiratory chain]. PMID- 3037852 TI - [Characterization of opioid receptors in the golden hamster brain]. PMID- 3037853 TI - [VIPergic transmitter of trachea smooth muscle in guinea pigs]. PMID- 3037855 TI - Afferent pathways of aortic baroreceptor fibers in guinea pigs. PMID- 3037854 TI - [Antagonism of the neuromuscular-blocking effect of polymyxin B by 3,4 diaminopyridine]. PMID- 3037856 TI - Effects of subacute treatment with toluene on cerebrocortical alpha- and beta adrenergic receptors in the rat. Evidence for an increased number and a reduced affinity of beta-adrenergic receptors. AB - Subacute treatment with toluene (80-1500 p.p.m.) produces a dose-dependent reduction of affinity and increase in density of the beta-adrenergic antagonist [3H]dihydroalprenolol binding sites in the frontoparietal cortex of the male rat, while the binding characteristics of alpha 1-adrenergic ([3H]WB 4101) and alpha 2 adrenergic ([3H]p-aminoclonidine) binding sites in the same region is unaffected by this treatment as evaluated in vitro. Therefore, it is suggested that the cortical beta-adrenergic receptors are particularly vulnerable to the action of toluene in vivo. It is speculated that as a result cortical beta-adrenergic neurotransmission may be altered following exposure to low concentrations of toluene, possibly related to the physico-chemical properties of toluene, leading to changes in membrane fluidity. PMID- 3037857 TI - Effects of calmodulin antagonists on hormone release and cyclic AMP levels in GH3 pituitary cells. AB - In GH3 cells the calmodulin antagonists trifluoperazine and N-(6-aminohexyl)-5 chloro-1-napthalene sulphonamide hydrochloride (W-7) showed a dose-dependent, biphasic effect on the release of growth hormone (GH) and prolactin (PRL). Hormone release was inhibited with 15-30 microM trifluoperazine and with 30-80 microM W-7, while stimulation was observed with 50-100 microM trifluoperazine and with 150 microM W-7. Trifluoperazine (greater than or equal to 30 microM) and W-7 (greater than or equal to 80 microM) increased the concentration of cellular cyclic AMP. Sulphoxides of trifluoperazine and chlorpromazine (less than or equal to 150 microM were without effect on hormone release and cellular cyclic AMP. Hydrolysis of cyclic AMP by GH3 cytosol was reduced after incubation of intact GH3 cells with trifluoperazine (15-60 microM). When trifluoperazine was incubated with cytosol, both the high and low affinity forms of cyclic AMP phosphodiesterase were inhibited competitively with calculated Ki of 4.5 and 56 microM, respectively. Stimulation of cyclic AMP phosphodiesterase caused by endogenous calmodulin was blocked by trifluoperazine. Particulate bound adenylyl cyclase activity was inhibited by trifluoperazine, and this effect was counteracted by endogenous calmodulin. PMID- 3037858 TI - Mapping of spinal projection of primary nociceptive neurones in the rat. AB - The FRAP (fluoride-resistant acid phosphatase) reaction was used to draw up the map of the somatotopic organization of the primary nociceptive neurones of the forelimb in the rat spinal cord. The disappearance of FRAP activity from the central terminals of primary sensory neurones in the substantia gelatinosa was studied after transection of the brachial plexus and the median nerve. A map of the central terminals of FRAP-reactive primary sensory neurones was drawn up by the recording of enzyme depletion. After complete transection of the brachial plexus, partial depletion of FRAP was seen in the lower cervical and upper thoracic cord; the degree of FRAP disappearance was striking in segments C5, C6 and C7, and slight between segments C5 and C6. After transection of the median nerve, the FRAP reaction was observed to disappear completely in the medial portion, but remained over approximately one-quarter of the width in the middle of the Rolando substance. The enzyme activity remained in a narrow strip about 2 segments long. The maps resulting from microscopic and geometric analysis of serial sections showed that the terminals of FRAP-reactive primary nociceptive neurones of the forelimb projected mostly to the medial portion of lamina II in the rat spinal cord, from segment C4 to T3. The findings are discussed in relation to the somatotopic organization of primary afferents. PMID- 3037859 TI - In vitro model for the response to irradiation of different types of human intracranial tumours. AB - Twenty-seven human low and high grade gliomas and five meningiomas were cultured in vitro as tumour tissue and/or tumour cells. Cell survival or growth was taken as a measure of radiation response. Astrocytomas II-III and glioblastomas manifested individual patterns of radiosensitivity, ranging from 10 to 90 Gy. Meningiomas did not react. Our findings are consistent with the differences in radiosensitivity of human gliomas experienced clinically and corroborate the validity of the in vitro model. PMID- 3037860 TI - [EEG and clinical study of West's syndrome. Factors with prognostic value]. PMID- 3037861 TI - Interdependency of Ca2+ availability and cyclic AMP generation in the pancreatic beta-cell. PMID- 3037862 TI - Induction of the glucokinase-glucose sensor in pancreatic islets of insulinoma bearing rats following tumor removal. PMID- 3037863 TI - A comparative study of cholecalciferol, dihydrotachysterol and alfacalcidol in the treatment of elderly patients with hypocalcaemia. AB - Fifty elderly patients with hypocalcaemia were randomly treated for 8 weeks with either oral dihydrotachysterol, parenteral cholecalciferol or oral alfacalcidol. All three treatments were successful in normalizing the serum calcium levels in most patients within 2 weeks. Hypercalcaemia was seen only with alfacalcidol but was rapidly reversed once treatment was discontinued. Hypercalcaemia was not observed with either dihydrotachysterol or cholecalciferol. These therefore, require less frequent biochemical monitoring. A single cholecalciferol injection eliminates the problems of compliance. PMID- 3037864 TI - Histamine dependent cAMP generating system in rabbit CNS: interaction with various neuroregulators. AB - Histamine (HI) potently stimulated, through H2-receptors, cAMP accumulation in slices of the rabbit cerebral cortex (EC50: 15 microM; maximum stimulation: 300 450% of the basal level). Combination of L-noradrenaline or L-adrenaline and adenosine with HI showed synergistic and additive effects, respectively. Carbachol, arecoline and pilocarpine had no effect, while oxotremorine and tremorine increased and decreased the HI response when used at low or high concentrations, respectively. PMID- 3037865 TI - Inhibition of sound-induced convulsions by metoprine in the audiogenic seizure susceptible rat. AB - Metoprine which increases brain histamine by blocking its methylation, was recently demonstrated to inhibit electrically induced tonic convulsions in rats. Its effect was now tested on audiogenic convulsions in genetically audiogenic seizure sensitive rats. Metoprine (20 mg/kg, i.p.) reduced the severity of seizures significantly 4 and 28 h after drug administration. Also the duration of convulsions was significantly decreased. These results agree with an involvement of histaminergic neurons in convulsive phenomena perhaps as a part of an anticonvulsive inhibitory transmitter system. PMID- 3037866 TI - Effect of a milk diet on rat gastric mucosa: receptor activity, histamine metabolism and ultrastructural analyses. AB - A bovine milk diet (BM) resulted in remarkable changes in histamine H2 receptor activity (sensitization) and PGE2 receptor activity (desensitization) in gastric glands isolated from adult rats. In contrast, the receptor-cAMP systems sensitive to glucagon(s) and secretin in parietal cells and muco-peptic cells were unaffected. In the two experimental groups, cimetidine produced a parallel displacement of the histamine dose-response curve suggesting competitive inhibition between this classical H2 receptor antagonist and histamine. The BM diet reduced the histidine decarboxylase activity in rat gastric mucosa; the histamine content was not significantly different in control and BM-fed rats. There was no alteration of the circadian rhythm of the parietal cell (ultrastructural changes: microvilli, tubulo-vesicles) determined at intervals of 6 hours in milk-fed rats. Prostaglandins and other components in milk (EGF, somatostatin, etc.) might therefore protect gastric mucosa by a differential control of PGE2 and histamine H2 receptor activity, either directly (PGE2 and EGF in milk) or indirectly (inhibition of endogeneous histamine synthesis/release and stimulation of prostaglandin synthesis/release). PMID- 3037867 TI - Interaction of cimetidine and histamine with superoxide generated in a cell-free system and in neutrophils. AB - We investigated the influence of CIM and HIS on the SUP and CH in the SUP generating cell-free xanthine oxidase system and in human neutrophils. As measured by CCR or CH, we found that HIS inhibited SUP output in both systems. The HIS concentration required for a measurable inhibition was somewhat higher in the xanthine oxidase system than in neutrophils. CIM increased CH in neutrophils significantly. A similar effect of CIM was not observed in the xanthine oxidase system. CIM antagonized in neutrophils the effect of HIS. However, a five or ten times higher molar concentration of CIM than of HIS was necessary to produce this effect. The antagonistic effect of CIM was not found in the xanthine oxidase system within the concentrations applied. In addition to an interaction of HIS with neutrophils on the cellular level, we suggest a superoxidase scavenging effect caused by HIS. At the present no suggestions can be made regarding the observed SUP output stimulating activity of CIM. PMID- 3037868 TI - [Seroepidemiological study of conjunctivitis related adenoviruses]. PMID- 3037869 TI - Nuclear and cytoplasmic maturation of bovine oocytes cultured with dbc AMP, FSH and hCG. PMID- 3037870 TI - In-vitro anti-microbial susceptibility of clinical isolates of pathogenic bacteria to ten antibiotics including phosphomycin. AB - The minimum inhibitory concentration (MIC) of ten antibiotics was determined for various bacterial pathogens isolated from clinical specimens in a Lagos hospital. The in-vitro activity of penicillin and tetracycline was not very impressive and a similarity was noticed in the resistance patterns of these two antibiotics, while the activity in vitro of the relatively more toxic aminoglycosides and chloramphenicol was high for Gram-negative rods. Ceftazidime demonstrated the highest activity in vitro against all pathogens studied. Cefoxitin, cefuroxime and phosphomycin demonstrated an impressive activity in vitro. Clindamycin was very active against strains of Staphylococcus aureus. beta-lactamase production amongst these strains was studied and the clinical significance of this and their MIC results are discussed. PMID- 3037871 TI - Cancer of the male breast: analysis of forty-three cases in Ibadan, Nigeria. AB - Male breast cancer accounts for 0.4% of all male neoplasia in Ibadan, and about 3.4% of all breast cancers in both females and males. The clinical features often presented include mass in breast, skin involvement by fixation and ulceration, sanguinous and serous discharge from the nipple. Symptoms are often delayed, the longest duration in the current series being 36 months. Geographical comparison of the histological types of male breast cancer appears to show that, while papillary carcinoma is commoner in Nigerian than in American and Danish males, on the other hand, infiltrating duct carcinoma appears much more common in the latter than in the former male races. Invasive lobular carcinoma occurs more in Nigerian than in American and Danish males. Mucinous carcinoma, on the other hand, appears much more common in the Danish than in the Nigerian and American males. In all the compared races, prognosis in male breast cancer appears to depend on the age at onset of the disease, the histological type, and presence or absence of lymph node metastases. In Nigerian males mucinous carcinoma appears to offer an excellent prognosis compared with papillary carcinoma, which has better prognostic features in American male patients. PMID- 3037872 TI - Prospective study of the pattern of utilization of mental health services of a Nigerian university hospital. AB - A prospective study of the pattern of utilization of the mental services offered by a Nigerian university teaching hospital was carried out in 1981. Demographic and other characteristics of the 151 patients registered over a 3-month period are described along with their mode of entry into the health care delivery system of the hospital. The significance and implication of the findings are discussed. PMID- 3037873 TI - Spontaneous contractile pattern of isolated circular and longitudinal muscle layers from the ampullae of normal and damaged fallopian tubes. AB - Ampullary biopsies were obtained from twenty-three normal fertile women and from eleven women with hydrosalpinges. The contractile activity of 4-mm muscle strips was studied isometrically in organ bath. Both normal circular and longitudinal muscle strips contracted with similar frequency, duration and amplitude. Circular and longitudinal muscle strips from hydrosalpinges contracted with a lower frequency and a longer duration and the amplitude of contractions was higher than normal. The difference in frequency and duration was statistically significant. The possible explanations for these differences in rhythmicity between normal and diseased fallopian tubes are presented and the prognostic relevance discussed. PMID- 3037874 TI - Normal range of lumbar spine flexion in adult Nigerians. AB - One hundred patients, fifty males and fifty females, without any back pain or disease, were studied. Their ages ranged between 14 years and 70 years. The lumbar flexion measurement was taken using the MacRae and Wright modified Schober's method. The values for males were higher than females and the younger the patient, the higher was the value of the lumbar flexion, up to the third decade. The method could be adopted to show improvements during treatment of low back pain by orthopaedic and physiotherapic practices. PMID- 3037875 TI - The pharmacokinetics of proguanil in human subjects following a single oral dose. AB - The pharmacokinetics of orally administered 200-mg dose of proguanil in volunteers, one African and one Caucasian, is described. The drug was rapidly absorbed reaching a peak concentration in the blood within 3 h, and declining slowly thereafter to give a terminal phase elimination half life of 11.20 +/- 4.10 h and a systemic clearance of 1.270 +/- 0.020 l/h/kg. The small apparent volume of distribution shows that the drug is confined mainly to the blood and is not extensively bound to tissues; it undergoes cyclic oxidation in the liver to cycloguanil--the active metabolite responsible for antimalarial activity. Cycloguanil was detected in the plasma 3 h after proguanil ingestion and reached peak concentration between 5 h and 6 h. Excretion of proguanil was rapid, 60% of the single dose passing through the renal system within 24 h. PMID- 3037876 TI - Smoking patterns in students of higher institutions of learning in Nigeria. AB - A study of the pattern of cigarette smoking among 2317 students of both sexes attending some selected universities, polytechnics, colleges of education and schools of nursing in Nigeria was undertaken by means of questionnaires. It was observed that 436 (30%) of male students and 174 (21%) of female students smoked cigarettes. The prevalence was noted to be on the increase for female students when compared with previous studies carried out in similar institutions in Nigeria. The highest prevalence of smoking was among students from colleges of education. Over 50% of the students were mild smokers while less than 50% had exceeded a duration of 5 years of smoking. The latter finding suggests that it may still be feasible to convince a large proportion of them to stop smoking or to cut down on the number of cigarettes smoked. PMID- 3037877 TI - Leukotriene antagonists: new antiasthmatic drugs. PMID- 3037878 TI - Why do the kidneys release renin in patients with congestive heart failure? A nephrocentric view of converting-enzyme inhibition. PMID- 3037879 TI - Severe hemorrhage from cytomegalovirus rectal ulcers in a burned adult. AB - Clinically evident gastrointestinal involvement by cytomegalovirus has been amply documented in various immunocompromised states with the notable exception of burns. A 44-yr-old man, having sustained 40% burn, who developed severe bleeding from rectal ulcers at the time of a primary cytomegalovirus infection is described. Evidence is provided, implicating cytomegalovirus as the responsible pathogen. We suggest that cytomegalovirus infection should be included in the differential diagnosis of gastrointestinal bleeding occurring in a burned patient. In that case, bleeding can occur from any part of the gastrointestinal tract. PMID- 3037880 TI - Digestibility of carbohydrate foods in an ileostomate: relationship to dietary fiber, in vitro digestibility, and glycemic response. AB - The aim of this study was to assess differences between starchy foods in the amount of carbohydrate which escapes small intestinal absorption. One ileostomate volunteer tested in metabolic feeding trials a total of 20 starchy foods (nine of which were repeated on two to seven occasions, mean 3.5 +/- 1.7). This ileostomate volunteer exhibited macronutrient and fiber losses that were within 97.3 +/- 6.6% of the mean, for a range of foods eaten by three other ileostomates and was therefore believed to be representative. Measurement of available carbohydrate in ileal effluent demonstrated a wide range of recoveries from 2.7 to 18% from different starchy foods. The available carbohydrate losses related to the fiber content (r = 0.885, p less than 0.001), in vitro digestibility of the food (r = -0.867, p less than 0.01), and their glycemic responses (r = -0.611, p less than 0.05). Our data support the concept that available carbohydrate losses to the colon may be greater in many foods than the carbohydrate entering the colon as dietary fiber. The food factors responsible are diverse and the possible metabolic consequences of carbohydrate malabsorption may be broad. PMID- 3037881 TI - Plasma levels of atrial natriuretic peptide in patients with chronic liver disease. AB - The plasma levels of atrial natriuretic peptide were determined by radioimmunoassay in 24 patients with chronic liver disease, including three patients with alcoholic liver disease, four with chronic active hepatitis, 13 with liver cirrhosis, and four with hepatocellular carcinoma. When compared with normal subjects (180 +/- 12 pg/ml), the plasma levels of atrial natriuretic peptide in cirrhotic patients (349 +/- 64 pg/ml) were significantly elevated (p less than 0.001) but not in other disease groups. In patients with chronic liver disease the plasma levels of atrial natriuretic peptide were correlated significantly with plasma renin activity but not with plasma aldosterone, and furthermore showed a negative correlation with indocyanine green disappearance rate. These results suggest that the increased plasma levels of atrial natriuretic peptide, which appear to be associated with an increase in plasma renin activity and with hepatic dysfunction, may participate in maintaining homeostasis of sodium and fluid volume in patients with chronic liver disease. PMID- 3037882 TI - Cytomegalovirus: a new gastrointestinal pathogen in immunocompromised patients. PMID- 3037883 TI - The spectrum of human immunodeficiency virus infection in patients with factor IX deficiency (Christmas disease) AB - Early reports suggested that hemophiliacs with factor IX deficiency (Christmas Disease) may be at less risk for developing the acquired immunodeficiency syndrome (AIDS) than patients with classic hemophilia. We evaluated 12 factor IX deficient patients for clinical and immunologic abnormalities related to infection with the human immunodeficiency virus (HIV). Antibody to HIV was not detected in these patients prior to 1982. By 1985, 66 percent (eight of 12) patients were seropositive. All three concentrates available commercially before 1985 were associated with seropositivity. Furthermore, seropositive hemophiliacs had received on average significantly more factor IX concentrate than seronegative hemophiliacs (27,825 +/- 17,976 (S.D.) versus 1,250 +/- 1,500 factor units/year, (p less than 0.02). Half of the seropositive individuals had generalized lymphadenopathy with splenomegaly. Two seropositive patients have developed AIDS, one with cryptococcal meningitis and another with a large cell immunoblastic lymphoma. Infection with HIV has occurred with high frequency in hemophiliacs who received unmodified factor IX concentrates. PMID- 3037884 TI - Hairy leukemic cells which hyperexpress Ii do not demonstrate Ii genome alterations by restriction endonuclease analysis. AB - The finding of increased expression and apparently altered processing of Ii in hairy leukemic cells led us to test for Ii genomic alteration by Southern-type Ii cDNA hybridization to leukemic spleen DNA cleaved with a series of restriction endonucleases. Some insertions, deletions, or point mutations, potentially detectable by this technique, might correlate to alteration in expression and function of Ii (and indirectly, class II antigens). No changes in genomic structure of Ii were detected in DNA isolated from spleens of five patients with hairy cell leukemia, compared with DNA preparations from peripheral blood cells of nineteen healthy blood donors. These experiments were consistent with the view that gross structural alteration of the Ii genome had not occurred in hairy leukemic cells. PMID- 3037885 TI - The kidney: therapeutic implications of ACE inhibition and other areas of new research. San Francisco, California, June 28-29, 1985, February 28-March 1, 1986. PMID- 3037886 TI - Effects of converting-enzyme inhibitors on the renin-angiotensin-aldosterone, bradykinin, and arachidonic acid-prostaglandin systems: correlation of chemical structure and biologic activity. AB - The renin-angiotensin-aldosterone system plays an integral role in the control of BP and is mediated by enzymatic transformation of substances in the renin angiotensin cascade. Inhibitors of various reactions in this cascade have been shown experimentally to reduce elevated BP. The first such compound to become available for clinical use was the angiotensin-converting enzyme inhibitor captopril. Captopril reduces BP, with the expected decrease in plasma angiotensin II concentration and increases in renin and angiotensin I levels, but there is also evidence for a non-renin-dependent mechanism of action. In particular, according to cell culture data, captopril stimulates an increase in the synthesis of vasodilatory prostaglandins. Because enalapril, another angiotensin-converting enzyme inhibitor, does not augment in vitro synthesis of prostaglandins, this effect may be unique to the chemical structure of captopril rather than inherent to all such inhibitors. Studies using animal models of hypertension support this hypothesis. Captopril may also influence prostaglandin synthesis by reducing inactivation of bradykinin. PMID- 3037887 TI - Angiotensin-converting enzyme inhibitors: considerations regarding proteinuria. AB - The effects of captopril on renal function are evaluated in hypertensive patients with impaired renal function previously entered in a worldwide surveillance study. For the overwhelming majority of patients, control of BP was accompanied by stable or improved renal function. To date, the major adverse renal reactions have been a somewhat increased risk of worsening proteinuria among these patients and the well-documented acute decrease in renal function that may occur in patients with bilateral renal artery stenosis. PMID- 3037888 TI - Renal abnormalities in nonmodulating essential hypertension. AB - Hypertensives with low renin levels are known to be sensitive to salt intake and diuretics. The subset of hypertensives sensitive to salt and having normal or high renin levels are termed nonmodulators. These are patients with essential hypertension who fail to modulate their renal blood flow and aldosterone responsiveness to angiotensin II when dietary sodium is changed. The result of nonmodulation is a pressor response to a short-term sodium load that results from a failure to excrete the sodium appropriately and that is presumably secondary to the defect in renal blood supply. On a high sodium load only nonmodulators have a decrease in BP in response to short-term treatment with an angiotensin-converting enzyme (ACE) inhibitor, which indicates an abnormality in intrarenal angiotensin II levels or in the renal angiotensin II receptor. PMID- 3037889 TI - Clinical experience with converting-enzyme inhibitors in hypertension. AB - Because the renin-angiotensin system frequently participates in sustaining severe forms of hypertension, early studies with the angiotensin-converting enzyme inhibitor captopril were concentrated in patients whose hypertension was refractory to previously available treatment. The effectiveness of captopril, both acutely and during long-term treatment, led to its widespread use for treatment-resistant hypertension. More recently, captopril's efficacy when used as monotherapy and its relative avoidance of symptomatic adverse effects have expanded its role to hypertension of all degrees of severity. A second converting enzyme inhibitor, enalapril, has been approved for the treatment of hypertension. Its efficacy and clinical characteristics appear similar to those of captopril. In double-blind clinical trials, comparisons with beta-blocker or diuretic therapy have shown captopril to be equal in efficacy but to produce fewer side effects or metabolic changes. In mild-to-moderate essential hypertension it is effective when administered in twice-daily doses of 25 or 50 mg; higher doses do not usually increase its BP-lowering effects. The combination of captopril with a diuretic in low doses has been shown to be especially effective. In additional studies, once-daily doses have controlled BP in a majority of hypertensive patients, but this method of administering captopril has not yet been formally approved. In lower doses, captopril may be less effective in black patients than in white patients, but black patients appear to respond well if the doses are increased. Antihypertensive effects in elderly patients during captopril treatment are equal to those in the young. Converting-enzyme inhibitor therapy appears to have broad utility and a favorable side effect profile in the treatment of essential hypertension. PMID- 3037890 TI - The treatment of renovascular hypertension: surgery, angioplasty, and medical therapy with converting-enzyme inhibitors. AB - Surgery has been used to treat renal vascular hypertension for almost 50 years. The reason for the many apparent discrepancies in the literature on effectiveness and risk have become clear only in the past decade. The results are poorest and the risk is greater, not surprisingly, in patients with advanced atherosclerosis involving many vascular beds. The results are much better in fibromuscular disease, both in terms of effectiveness and risk. Angioplasty has been available for a much shorter time, but a reasonable picture of the short-term effectiveness and the risk is emerging. The risk is substantially lower than that of surgery. The results are again best in fibromuscular disease. In atherosclerotic disease, the results are especially poor for the most common lesion, that involving the renal artery ostium. Medical therapy before the development of captopril was often difficult and often unsatisfactory. Since the development of converting enzyme inhibition, medical therapy is an important option. In the early experience, reflecting the severity of the hypertension, the frequency with which azotemia was present, and the high dose of captopril used, the adverse reaction rate was substantial. In one study, none of 133 patients with unilateral renal arterial disease and an intact contralateral kidney developed renal failure. Among 136 patients with bilateral disease or a solitary kidney, renal failure occurred in 15 and led to discontinuation of therapy in 12. If surgery or angioplasty are contraindicated, one can modify the therapeutic goal.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3037891 TI - Adaptive and maladaptive actions of angiotensin II in patients with severe congestive heart failure. AB - The renin-angiotensin system appears to have evolved millions of years ago as a primary attempt to preserve circulatory homeostasis at a time when the principal cause of a low cardiac output was intravascular volume depletion. Angiotensin II supported systemic BP by direct systemic vasoconstriction, by facilitating the central and peripheral effects of the sympathetic nervous system, by promoting renal sodium retention by the production of aldosterone, and by increasing total body water by enhancing thirst and the synthesis of vasopressin. In addition, angiotensin II evolved as an important mechanism to preserve the glomerular filtration rate in low-flow states. These actions of angiotensin II were beneficial when the system first evolved, but its activation in patients with heart failure not only fails to reverse the low-output state but further exacerbates loading conditions in the left ventricle, thereby leading to worsening heart failure. Moreover, increased levels of angiotensin II cause heightened sympathetic nervous activity, potassium depletion, and hyponatremia, each of which can lead to further clinical deterioration. Therefore, activation of the renin-angiotensin system in heart failure might appear (at first) to be a maladaptive response. Recent evidence, however, suggests that this hormonal system continues (even in heart failure) to carry out the primary functions for which it was designed. Angiotensin II plays an important role in preserving systemic BP and in preserving the glomerular filtration rate as renal artery pressure and renal blood flow decline; in addition, by stimulating the synthesis of aldosterone, the renin-angiotensin system provides an important role for potassium disposal.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3037892 TI - Atrial natriuretic peptide and the kidney. AB - Studies addressing the mechanisms of atrial natriuretic peptide (ANP) action within the kidney are reviewed. The magnitude of the natriuretic response to ANP initially suggested inhibition of renal sodium transport. It now appears, however, that the renal response to ANP is largely dependent on ANP-induced alterations in renal hemodynamics. At the glomerulus, ANP-induced afferent arteriolar dilatation and efferent arteriolar constriction produce a rise in the glomerular capillary hydraulic pressure and thus an increase in the filtration fraction and glomerular filtration rate. Furthermore, angiotensin II-induced increments in renal perfusion pressure markedly augment ANP-induced NaCl excretion, whereas increments in peritubular oncotic pressure or reductions in renal perfusion pressure nearly abolish this natriuretic effect, thereby suggesting that changes in the peritubular physical factors, which govern tubule fluid reabsorption, are required to elicit the natriuretic action of ANP. In addition, increments in papillary vasa recta hydraulic pressures during ANP infusion argue for an important influence of ANP on fluid exchange in the renal papilla. There also is recent evidence to suggest that ANP directly inhibits papillary collecting duct NaCl reabsorption. This action of ANP is thought to contribute to the enhanced renal excretion of sodium-rich urine in response to ANP. Finally, in two models of chronic extracellular volume expansion, namely, mineralocorticoid excess and a reduction in nephron number with a high sodium intake, endogenous plasma ANP levels increased significantly above control levels.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3037893 TI - Renal effects of angiotensin-converting enzyme inhibition in cardiac failure. AB - Compensatory mechanisms such as systemic vasoconstriction and sodium retention are activated to various degrees in patients with congestive heart failure (CHF). The renin-angiotensin system (RAS) mediates many of these compensatory responses. Increased angiotensin can result in net vasoconstriction and increase afterload. In addition, it contributes to the avid sodium retention that occurs in patients with CHF. The RAS promotes sodium retention by enhancing proximal tubular reabsorption of sodium and water through changes in intrarenal hemodynamics. In addition, the direct renal tubular influence of angiotensin II and aldosterone contribute to sodium retention. In experimental heart failure in the rat, angiotensin activation leads to reduction in effective plasma flow whereas the glomerular filtration rate was unchanged. In clinical CHF, activation of the RAS may also contribute to azotemia in patients with CHF. Significant correlations between serum creatinine levels and plasma renin activity have been observed. Furthermore, renal plasma flow and the glomerular filtration rate frequently improve after therapy with converting-enzyme inhibitors (CEIs). Occasionally, however, CEIs may produce an excessive fall in systemic BP and, hence, renal perfusion pressure, which may overcome autoregulatory mechanisms and contribute to a deterioration in renal function. CEIs enhance the effectiveness of furosemide in promoting natriuresis and correction of hyponatremia. Diuretic induced hypokalemia can also be corrected by angiotensin-converting enzyme (ACE) inhibitors due to blockade of aldosterone production. On the basis of these results, we recommend titration of the ACE inhibitors' dose to minimize hypotension and the concurrent use of furosemide in patients with CHF receiving ACE inhibitors, especially for those in whom azotemia, hyponatremia, and hypokalemia complicate the course. PMID- 3037894 TI - Therapeutic implications of converting-enzyme inhibitors in renal disease. AB - Micropuncture and morphological studies were performed in three protocols assessing the renal hemodynamic and structural effects of angiotensin I converting enzyme inhibitors (CEIs) in the progression of glomerular injury. In protocol I, rats were subjected to 5/6 renal ablation and received no therapy, enalapril (CEI), or triple-drug therapy (TRX) for 12 weeks. Control of systemic and glomerular hypertension with CEI resulted in prevention of glomerular capillary hypertension and protection against glomerular injury. Despite equivalent control of systemic BP, failure of TRX to control glomerular hypertension was associated with no protection against eventual proteinuria and glomerular sclerosis, values for these indexes being as abnormal as in rats receiving no therapy. In protocol II, rats were again subjected to 5/6 renal ablation and followed for 18 weeks. Early institution of CEI soon after ablation again prevented systemic and glomerular hypertension and largely limited glomerular injury. In a third group, enalapril therapy was delayed for 8 weeks after ablation until hypertension and proteinuria were established. Late institution of CEI resulted in prompt reduction in systemic and glomerular capillary hypertension and stabilization of glomerular disease. In protocol III, CEI was administered to normotensive, moderately hyperglycemic diabetic rats. A modest, 20-mm Hg reduction in systemic arterial pressure was associated with the normalization of glomerular capillary pressure and a striking reduction in the development of albuminuria and glomerular injury. These studies suggest that CEI effectively prevent glomerular capillary hypertension and thereby afford protection against glomerular injury in diverse models of progressive renal disease. PMID- 3037895 TI - Occupational factors associated with astrocytomas: a case-control study. AB - The most malignant form of all brain tumors is the supratentorial astrocytoma. Little is known about its etiology, but exogenous factors have been blamed. In this case-control study, 78 astrocytoma patients have been compared with 197 clinical and 92 population controls. An extensive questionnaire was used to gather information about occupational and residential environment exposure. Inquiries concerning groups of or individual chemicals elicited low rates of affirmative response, with negligible differences between cases and controls. However, the questions "working at an airfield" and "living near a petrochemical plant" indicated elevated risks in comparison with both control groups; so too did "living near a municipal sewage treatment plant." These results focus attention on exposure to organic compounds and should be considered together with similar findings in current research. No other occupation, branch of industry, or vicinity questions showed differences between cases and controls, with the exception of "living in the neighborhood of a paper mill or a saw mill," which gave moderately increased relative risks. A separate report gives the results from the nonoccupational part of the study. PMID- 3037896 TI - Influence of diabetes mellitus on changes in left ventricular performance and renal function produced by converting enzyme inhibition in patients with severe chronic heart failure. AB - Diabetes mellitus is frequently accompanied by specific abnormalities of the renin-angiotensin system, but it is not known whether these alterations modify the response to converting enzyme inhibition. To evaluate this possibility, 129 patients with severe chronic heart failure were treated with captopril or enalapril for one to three months, while doses of digoxin and diuretics were kept constant; 35 patients had diabetes mellitus. Prior to therapy, diabetic patients had lower plasma renin activity (3.4 +/- 0.5 versus 7.0 +/- 1.1 ng/ml/hour) than did nondiabetic control subjects (p less than 0.05); yet the initial hemodynamic response to captopril was similar in both groups. Plasma renin activity predicted the hypotensive response to the first dose of captopril in nondiabetic control subjects (r = 0.70, p less than 0.001) but not in diabetic patients (r = 0.29). During long-term treatment with captopril or enalapril, both diabetic and nondiabetic patients had similar increases in cardiac index and decreases in mean arterial pressure and systemic vascular resistance. Diabetic patients, however, showed larger reductions in left ventricular filling pressure (-13.8 versus -9.1 mm Hg, p less than 0.02) and mean right atrial pressure (-6.2 versus -3.9 mm Hg, p less than 0.05) than did nondiabetic subjects; this was accompanied by a notable decline in body weight in diabetic patients only. Renal function remained unaltered during converting enzyme inhibition in nondiabetic patients, but deteriorated significantly in diabetic patients, as reflected by a marked increase in serum creatinine concentration (1.7 +/- 0.1 to 2.1 +/- 0.1 mg/dl, p less than 0.001). In conclusion, despite lower pretreatment plasma renin activity, diabetic patients with severe chronic heart failure demonstrated improvement during long-term converting enzyme inhibition to a degree similar to (if not greater than) that seen in nondiabetic control subjects, but were more susceptible to the development of functional renal insufficiency than their nondiabetic counterparts. These differences are explicable by abnormalities of renin/aldosterone synthesis and angiotensin-mediated vasoregulation that are known to be present in the diabetic state. PMID- 3037897 TI - Hypercalcemia in breast cancer. Reassessment of the mechanism. AB - Hypercalcemia in patients with breast cancer is usually attributed to osteolytic bone metastases. Seventeen patients with biopsy-proved breast cancer and hypercalcemia were identified in a prospective, unselected manner. Biochemical and clinical evaluation included measurements of parathyroid hormone, nephrogenous cAMP, vitamin D metabolites, fasting calcium excretion, and maximal tubular phosphate reabsorption, and bone radionuclide scanning. Tumor histologic findings were also reviewed. Four of the 17 patients (23.5 percent) had no evidence of bone involvement by bone scanning or radiography. Two additional patients (a total of 35 percent) appeared to have a humoral component to their hypercalcemia as determined by the presence of elevated nephrogenous cAMP excretion. These observations suggest that humoral, tumor-derived products may play a more important role in the hypercalcemia of breast cancer than has been previously recognized. PMID- 3037898 TI - Syncope and hypotension due to carcinoma of the breast metastatic to the carotid sinus. AB - Tumors involving the carotid sinus and glossopharyngeal nerve may produce syncope due to bradycardia and hypotension. Carotid sinus syncope unrelated to cancer is usually caused by bradycardia and responds to control of the heart rate. When neoplastic disease involves the carotid sinus, vasodepressor hypotension, with or without bradycardia, is more common. Control of the heart rate alone is not effective. Although this syndrome is not common, it is probably not recognized in milder forms. Most patients in whom this syndrome develops have cancer of the head and neck. A patient with breast carcinoma metastatic to the neck and carotid sinus is described in whom syncope with hypotension and bradycardia developed. Although a temporary cardiac pacemaker controlled bradycardia, severe hypotensive episodes recurred despite treatment with anticholinergic and sympathomimetic drugs. The pathophysiology and therapy of this syndrome in patients with cancer are reviewed. PMID- 3037899 TI - Norfloxacin for prevention of bacterial infections in granulocytopenic patients. AB - The efficacy and safety of norfloxacin were compared with those of placebo, vancomycin-polymyxin, and trimethoprim-sulfamethoxazole (TMP/SMX) for prophylaxis of bacterial infections in granulocytopenic patients. The study results showed that norfloxacin treatment, which was well tolerated and not associated with any serious systemic adverse effects, prevented acquisition of gram-negative bacillary organisms. Fewer norfloxacin-treated patients (38 of 108 patients, or 35 percent) experienced microbiologically documented infections compared with patients receiving placebo (27 of 40 patients, or 68 percent), vancomycin polymyxin (16 of 30 patients, or 53 percent), or TMP/SMX (14 of 28 patients, or 50 percent). Gram-negative bacteremia developed in five of 108 norfloxacin treated patients (5 percent), compared with 17 of 40 placebo-treated patients (43 percent), five of 30 treated with vancomycin-polymyxin (17 percent), and one of 28 patients treated with TMP/SMX (4 percent). The incidence of gram-positive bacteremia was similar in all study groups and was not affected by norfloxacin or any other oral prophylactic antibiotics. These results suggest that norfloxacin is both safe and effective for the prevention of serious gram-negative bacillary infections in granulocytopenic patients. More effective prophylaxis of gram positive bacterial infections, however, is needed. PMID- 3037900 TI - Hypouricemia in liver cell carcinoma: an isolated renal tubular defect. PMID- 3037901 TI - Pneumonias in adults due to mycoplasma, chlamydiae, and viruses. AB - Pneumonias in adults due to mycoplasma, chlamydiae, and viruses are a common clinical problem. These microorganisms contribute to the etiologies in 6-35% of all cases of pneumonia and are the sole pathogens in 1-17% of hospitalized cases. Important trends and developments in the field include the emergence of a Chlamydia psittaci strain (TWAR) that is passaged from human to human, causes a mycoplasma-like illness, and that is relatively resistant to erythromycin, the recognition of respiratory syncytial virus as a pathogen in nursing home outbreaks and in immunosuppressed adults, the continuing high lethality of fully developed influenza pneumonia, the efficacy of acyclovir and adenine arabinoside in limiting the complications of varicella-zoster virus infections, and the increasing frequency of pneumonia caused by cytomegalovirus and the severity of this disorder in highly immunosuppressed patients. Developments in the rapid diagnosis and therapy of respiratory syncytial virus infections with an aerosolized antiviral drug in children may pave the way for comparable advances in difficult pneumonias in adult patients. PMID- 3037902 TI - Nasopharyngeal carcinoma, aplastic anemia, and various malignancies in a family: possible role of Epstein-Barr virus. AB - We report on a family ascertained because of sibs affected with nasopharyngeal carcinoma. Other relatives had aplastic anemia or other malignancies and most were born of consanguineous parents. Epstein-Barr virus has been strongly implicated in the pathogenesis of nasopharyngeal carcinoma. It has also been suggested as a cause of aplastic anemia and other neoplasms. We postulate that this extensive German-Mennonite family has a heritable defect in the immune response to Epstein-Barr virus making its members at excessive risk for a number of different malignancies. This immunologic predisposition may be HLA associated and appears to be inherited as an autosomal recessive trait. PMID- 3037903 TI - Trisomy 18 and hepatic neoplasia. AB - A 2 9/12-year-old girl with trisomy 18 presented with a 3-week history of low grade fever, abdominal distention, and hepatosplenomegaly. Abdominal cytotomography (CT) scan showed hepatic infiltration with a tumor mass presumed to be hepatoblastoma. She deteriorated rapidly and died 3 weeks later. No autopsy and/or biopsy could be done. PMID- 3037904 TI - Cleft lip/palate-oligodontia-syndactyly-hair alterations, a new syndrome: review of the conditions combining ectodermal dysplasia and cleft lip/palate. AB - We report on a noninbred girl with cleft lip and palate, complete absence of deciduous teeth, hypodontia of permanent teeth, hair alterations, hypertelorism, midface hypoplasia, abnormal EEG, syndactyly, and other findings. Her mother had minor anomalies which could represent the mild expression of a gene. A review on the conditions combining ectodermal dysplasia and cleft lip/palate is presented. PMID- 3037905 TI - Onset in the seventh decade and lack of symptoms in heterozygotes for the TTRMet30 mutation in hereditary amyloid neuropathy-type I (Portuguese, Andrade). AB - In a Portuguese-American family with hereditary amyloid neuropathy (familial amyloidotic polyneuropathy), onset was in the seventh decade in all affected relatives. Another unusual characteristic was their origin from the Portuguese island of Madeira. In spite of this, the mutant transthyretin (TTRMet30) (the same variant prealbumin that is the circulating precursor of AFP protein in the classic Portuguese patients) could be found in the propositus' plasma. In addition, three other asymptomatic relatives (ages 90, 73, and 48) were shown to carry the mutation. Late onset and incomplete penetrance, at a clinical level, raise problems for presymptomatic detection of mutant TTR, as these tend to cluster in families. When counseling asymptomatic heterozygotes, we must consider intra-familial correlation in age-of-onset, and the distribution of age-of-onset including age of unaffected heterozygotes. This family poses interesting questions regarding pathogenesis of this degenerative process and the influence of other genetic factors, such as modifiers, epistasis, and polymorphism of the TTR genes or their regulators. A cis-effect of a gene linked to the mutant gene, decreasing the synthesis of the mutant TTR and keeping a sufficient amount of the normal one in circulation, or producing some cofactor for TTR, could also explain late onset and apparently incomplete penetrance; the occasional finding of classic forms in these families would be the result of recombinatory events. PMID- 3037906 TI - Mechanisms and regulation of renal H+ and HCO3- transport. PMID- 3037907 TI - Effects of naloxone administration on endocrine abnormalities in chronic renal failure. AB - In 72 patients with end-stage renal failure and 70 healthy subjects, the influence of blockade of opioid receptors by naloxone on secretion of prolactin, lutropin (LH), follitropin (FSH), adrenocorticotropin (ACTH), somatotropin (HGH), insulin (IRI), glucagon (IR-G), parathyroid hormone (PTH) and calcitonin (CT) was studied. Administration of naloxone stimulated luliberin-induced LH and FSH secretion quantitatively equally in patients and controls. Blockade of opioid receptors was followed by a less marked suppression of chlorpromazine-induced prolactin secretion but by a higher response of hypoglycemia-induced ACTH secretion in uremic patients than in controls. In addition, a less marked suppressive effect of naloxone was noted on hypoglycemia-induced HGH secretion in chronic renal failure as compared with controls. Blockade of opioid receptors improved significantly glucose tolerance and glucose-induced insulin secretion in uremic patients and suppressed nearly completely glucagon secretion response during the second phase of a glucose tolerance test. Finally, administration of naloxone was followed by a blunted response of Ca-induced CT secretion and suppression of PTH. Data presented in this paper suggest the existence of hyperendorphinism in end-stage renal failure. PMID- 3037908 TI - Diet and cancer--what are the links? PMID- 3037909 TI - Proopiomelanocortin-related peptides and methionine enkephalin in human follicular fluid: changes during the menstrual cycle. AB - Several studies indicate the presence of different pituitary hormones or neuropeptides in ovarian follicular fluid from various species. Recently our group showed that the ovarian follicular fluid of health women contains two of the endogenous opioid peptides, beta-endorphin and methionine enkephalin, in concentrations that are tenfold to twentyfold higher than in circulating plasma. The presence of immunoreactive beta-lipotropin was also shown. The aim of the present study was to evaluate whether adrenocorticotropic hormone, which in pituitary cells is synthesized from proopiomelanocortin such as beta-endorphin and beta-lipotropin, is also present in follicular fluid and the possible changes of proopiomelanocortin-related peptides during the menstrual cycle. Concentrations of beta-endorphin, methionine enkephalin, adrenocorticotropic hormone, and beta-lipotropin were measured in 60 healthy menstruating women at different periods of the menstrual cycle (20 during the follicular, 22 in the preovulatory days, and 18 during the luteal phase). Thirteen women participated in an in vitro fertilization program and thus received clomiphene citrate (100 mg/day from the fifth to the ninth day) plus 5000 IU human chorionic gonadotropin before starting the program. All samples were collected at laparoscopy under general anesthesia. In another eight patients fluid was collected from follicular cysts. Peptides were extracted on octadodecasilyl silica columns with 80% methanol in 0.5 mol/L acetic acid. The identity of follicular fluid beta endorphin, methionine enkephalin, adrenocorticotropic hormone, and beta lipotropin and standard peptides was demonstrated with high-pressure liquid chromatography. Peptide concentrations were measured in extracts by radioimmunoassays either directly by (methionine enkephalin and adrenocorticotropic hormone) or after (beta-endorphin and beta-lipotropin) gel filtration on Sephadex G-75. The concentrations of methionine enkephalin, adrenocorticotropic hormone, and beta-lipotropin were similar in the different periods of the cycle. Conversely, beta-endorphin concentrations were significantly higher in preovulatory days than in the other periods; no differences were evident between spontaneous and stimulated cycles. These results indicate that proopiomelanocortin-related peptides are present in the follicular fluid and that beta-endorphin concentrations change during the menstrual cycle, with the highest values occurring in the preovulatory follicle. PMID- 3037910 TI - Lymphadenopathy in macaques experimentally infected with the simian immunodeficiency virus (SIV). AB - A T-cell tropic lentivirus of macaques the simian immunodeficiency virus (SIV), has morphologic, growth, and antigenic properties that indicate that it is related to the human immunodeficiency virus (HIV), the etiologic agent of the acquired immune deficiency syndrome (AIDS) in humans. Six juvenile macaques developed persistent lymphadenopathy (greater than 3 months in duration) after inoculation with SIV. The histologic appearance of the lymph nodes was characterized by marked follicular hyperplasia with abundant proliferative B cells infiltrating into the paracortex. The number of T8-positive lymphocytes equaled or exceeded the number of T4-positive lymphocytes in the paracortex. These findings, in association with immunologic abnormalities and a previously observed fatal immunodeficiency syndrome in SIV-infected macaques, provide further evidence of the importance of SIV-induced disease in macaques as a model for the study of AIDS. PMID- 3037911 TI - Pancreas transplantation. An immunohistologic and histopathologic examination of 100 grafts. AB - The authors examined tissues obtained by biopsy, pancreatectomy, and autopsy from 100 pancreas grafts to determine the cause of dysfunction or failure of the graft. Immunohistologic examination of 42 tissues to determine the mononuclear cell phenotypes and Class I and II antigen expression was performed as well. Technical factors--infections, thrombosis, obstruction--accounted for a large number of graft losses, but immunologic-mediated mechanisms resulted in graft dysfunction and failure as well. Pleomorphic inflammatory infiltrates were present in grafts with acute rejection, as well as Silastic and Prolamine duct obstructed grafts. Criteria useful in the identification of acute rejection from pancreatitis included a more intense, predominantly mononuclear cell infiltration of transformed lymphocytes in the exocrine pancreas and evidence of vascular rejection--endovasculitis or fibrinoid necrosis. Increased expression and/or induction of Class I and II antigens on pancreatic constituents occurred in grafts with evidence of acute rejection, but also with Silastic and prolamine duct-obstructed pancreatitis. An isletitis occurred in 25% of the grafts. Nine of the 25 grafts (36%) with isletitis also had selective loss of beta cells from the islets. Recurrent diabetes mellitus appeared to have developed in these cases, which accounted for loss of graft function. PMID- 3037913 TI - Na-H and Cl-HCO3 exchange in rabbit oxyntic cells using fluorescence microscopy. AB - We have used the fluorescence intensity ratio (excitation 490/439; emission 520 550 nm) of 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein (BCECF) to measure intracellular pH (pHi) in single oxyntic cells (OCs) within intact gastric glands isolated from rabbits. The fluorescence ratio was converted to pHi by exposing cells to high [K] Ringer plus 10(-5) M nigericin. Ratios varied linearly with pHi over the physiological range. When the bathing solution was changed from NaCl to Na gluconate Ringer, pHi increased from 7.0 to 7.4. The increase of pHi occurred equally rapidly in nominally CO2-HCO3-free solutions and in solutions containing 5% CO2 and 25 mM HCO3. This effect was reversible and blocked by 2 X 10(-4) M 4,4'-diisothiocyano-2,2'-disulfonic acid stilbene (DIDS). The DIDS block was bypassed by 10(-5) M tri-n-butyltin, a pharmacological Cl-OH exchanger. When the bathing solution was changed from NaCl to N-methyl-D-glucamine (NMG) Cl Ringer, pHi decreased reversibly from 7.0 to 6.8. It was also found that changing from NMG gluconate to Na gluconate Ringer caused pHi to increase from 7.1 to 7.3, and this alkalinization was blocked by 10(-3) M amiloride; changing from NMG gluconate to NMG Cl Ringer caused pHi to decrease to 6.7. We conclude that OCs contain separate Na-H and Cl-HCO3 exchangers. Similarities and differences between these exchangers and those present in other cell types are discussed. PMID- 3037914 TI - Interactions of sodium-proton exchange mechanism in dog red blood cells with N phenylmaleimide. AB - Dog red blood cells (RBC) have a Na-H exchanger that is reversibly activated by cell shrinkage. The Na-H exchanger can be fixed in the on or off mode by treating the cells with N-phenylmaleimide. This action depends on the volume of the cells at the time of exposure to N-phenylmaleimide and also on the concentration of the reagent per number of cells. If the cells are swollen in hypotonic media during N phenylmaleimide exposure, the Na-H exchanger becomes irreversibly inactivated, so that on subsequent shrinkage of the cells, no amiloride-sensitive Na flux is seen. This effect is maximal at N-phenylmaleimide concentrations of greater than 20 mumol/g hemoglobin. If the cells are shrunken in hypertonic media during N phenylmaleimide exposure, the response of the Na-H exchanger depends critically on the concentration of the reagent. At N-phenylmaleimide concentrations of less than 20 mumol/g hemoglobin, the Na-H exchanger is fixed in the activated state, so that even when the volume stimulus is removed by subsequent cell swelling, an amiloride-sensitive flux is seen. Higher concentrations of N-phenylmaleimide applied to shrunken cells inhibit the Na-H exchanger. The results are accounted for in a model that envisions a volume-responsive switching mechanism for Na-H exchange that has two functional groups capable of reacting with N phenylmaleimide. The accessibility of these groups is determined by cell volume. PMID- 3037912 TI - Modulation of acute immune complex-mediated tissue injury by the presence of polyionic substances. AB - Considerable attention has been focused on the role of electrostatic charge in the pathogenesis of immune complex-mediated tissue injury. The authors have examined the ability of cationic (histone, polyhistidine, polyarginine) and anionic (polyanetholsulfonate) polyelectrolytes to modulate acute immune complex mediated tissue injury. Tissue injury elicited in rats by the reversed dermal Arthus reaction was increased 26-43% by addition of polyelectrolytes to antibody prior to its intradermal injection. Kinetic studies using 111In-labeled neutrophils indicated that the enhanced tissue injury was not the result of increased influx of neutrophils. Infusion of polyethylene glycol-conjugated superoxide dismutase prior to induction of the Arthus reaction resulted in 40-68% suppression of tissue injury. Concomitant in vitro functional studies (enzyme secretion, O-2 and H2O2 generation, and chemiluminescence) of rat neutrophils demonstrated that addition of polyelectrolytes to preformed immune complexes (IgG bovine serum albumin) resulted in marked increases in O-2, H2O2, and chemiluminescence, but no increases in enzyme secretion, compared with neutrophils stimulated with immune complexes alone. The cationic polyelectrolytes did not alter the capacity of preformed immune complexes to activate complement in vitro. These studies suggest that both cationic and anionic polyelectrolytes can increase the pathogenic potential of immune complexes and that this modulation is, at least in part, mediated by enhanced generation of toxic oxygen derived metabolites by neutrophils. PMID- 3037915 TI - Differences in CuZn superoxide dismutase induction in lungs of neonatal and adult rats. AB - The failure of adult rats to survive prolonged exposure to greater than 95% O2 is generally ascribed to the inability of their lungs to increase antioxidant enzyme synthesis in response to the oxidant challenge. We studied the synthesis rate of the antioxidant enzyme CuZn superoxide dismutase (CuZn SOD) in lungs of adult and neonatal rats exposed to conditions that alter the lung's oxidant-to-antioxidant balance. Lung CuZn SOD synthesis in the adult was significantly increased after 24 h of hyperoxia but fell to control levels after further exposure, whereas in neonatal lungs an increased rate of synthesis of CuZn SOD was found only after 72 h of hyperoxia. The adult lung responded to two in vitro oxidant stresses, [diethyldithiocarbamate exposure and heat (42 degrees C)] with increases in CuZn SOD synthesis twice the magnitude of those in the neonatal lung. These data indicate that the adult lung is at least as capable as the neonatal lung of increasing its synthesis of CuZn SOD in response to an oxidative stress. However, the inability of the adult lung to maintain an increased rate of CuZn SOD synthesis during in vivo hyperoxia may contribute to the poor tolerance of the adult lung to greater than 95% O2. PMID- 3037916 TI - Lipid transport function of lipoproteins in blood plasma. AB - Fatty acid and cholesterol transport in plasma lipoproteins evolved in the context of an open circulatory system in which lipoprotein particles are secreted directly into the blood and have ready access to cells in various tissues. In higher vertebrates with closed capillary beds, hydrolysis of triglycerides at capillary surfaces is required for efficient uptake of their component fatty acids into cells. Likewise, hydrolysis of cellular triglycerides in cells of adipose tissue precedes mobilization of the fatty acids and permits large amounts to be transported in the blood. However, in all Metazoa lipoproteins are secreted primarily from cells adjacent to an open microvascular bed. Uptake of lipoprotein particles as such into cells occurs in invertebrates and vertebrates alike, facilitated by binding to high-affinity receptors on cell surfaces. In vertebrates, a concentration gradient created between cholesterol in cells and lipoproteins by a cholesterol-esterifying enzyme that acts on lipoproteins promotes movement of cholesterol into the plasma compartment. Thus the strategies to transport poorly soluble lipids include enzymatic reactions at cell surfaces and in blood plasma as well as the processes of exocytosis and endocytosis. PMID- 3037917 TI - Parathyroid hormone acutely elevates intracellular calcium in osteoblastlike cells. AB - Changes in cytoplasmic calcium concentration ([Ca2+]i) activate numerous cellular processes thus mediating the effects of a number of hormones, but whether this mechanism is involved in the activation of osteoblasts by parathyroid hormone (PTH) remains uncertain. To examine this question, [Ca2+]i has been measured in suspensions of UMR 106 cells, a rodent osteosarcoma cell line with an osteoblastic phenotype. Basal [Ca2+]i was 137 +/- 3.7 nM (n = 60) and after the addition of rat PTH-(1-34) [rPTH-(1-34)] there was a rapid, dose-related increase with return to base line within 1 min. Half-maximal stimulation was produced by 5 X 10(-8) M rPTH-(1-34). Complexing of intracellular calcium by EGTA addition immediately before that of rPTH did not affect the calcium transient; neither did MnCl2 (10(-4) M) nor diltiazem (10(-4) M). Verapamil (10(-5) M) reduced the [Ca2+]i peak height after rPTH to 0.48 +/- 0.14 of control (n = 7). 8-(N,N diethylamino)octyl-3,4,5-trimethoxybenzoic acid and dantrolene both reduced the [Ca2+]i response to rPTH (0.65 +/- 0.08 and 0.29 +/- 0.13 of control, respectively). Forskolin (10(-6) and 10(-5) M) produced a slight [Ca2+]i transient smaller in amplitude than seen with PTH. It is concluded that PTH mobilizes an intracellular calcium pool in these osteoblastlike cells, and the predominant mechanism for this is independent of cAMP. PMID- 3037918 TI - A role for iron in oxidant-mediated ischemic injury to intestinal microvasculature. AB - Recent reports in the literature suggest that iron plays an important role in free radical-mediated injury in biological systems. To assess the role of iron catalyzed oxidant production in ischemia-reperfusion injury, we examined the influence of deferoxamine (an iron chelator) and apotransferrin (iron transporting protein) on the increased intestinal vascular permeability produced by 1 h of ischemia and reperfusion. Both agents were administered intravascularly as a constant infusion, beginning 5 min before reperfusion. Capillary osmotic reflection coefficients were derived from the relationship between lymph-to plasma protein concentration ratio and lymph flow in the feline small bowel. Vascular permeability in control intestinal preparations was 0.08 +/- 0.005, however it increased significantly to 0.40 +/- 0.03 in preparations subjected to 1 h of ischemia and 30 min of reperfusion. Vascular permeability in the deferoxamine-(0.15 +/- 0.009) and apotransferrin- (0.17 +/- 0.002) treated animals were significantly lower (P less than 0.01) than in the untreated group. Treatment with iron-loaded deferoxamine or transferrin did not offer any protection against ischemic injury. These findings support the hypothesis that iron plays an important role in the formation of hydroxyl radicals after reperfusion of the ischemic bowel. PMID- 3037919 TI - Cellular action of arginine vasopressin in the isolated renal tubules of hypothyroid rats. AB - Hypothyroidism has been demonstrated to be associated with an impaired concentrating capacity and specific morphological changes in the thick ascending limbs. This study was performed to evaluate the cellular action of arginine vasopressin (AVP) in the isolated renal tubules from control (C) and hypothyroid (HT) rats. Hypothyroidism was induced by feeding aminotriazole for 4 wk. Urinary volume was higher in HT rats (C 13.5 +/- 0.9, HT 17.7 +/- 0.9 ml/24 h, P less than 0.005) and urinary osmolality was lower in HT rats (C 1,707 +/- 49, HT 1,229 +/- 35 mosmol/kgH2O, P less than 0.001). Plasma AVP levels were significantly higher in HT rats (C 1.93 +/- 0.59, HT 4.12 +2- 0.62 pg/ml, P less than 0.05), thus documenting AVP resistance. The adenylate cyclase response to AVP (10(-6) M) was significantly lower (P less than 0.02) in the medullary thick ascending limb of Henle's loop (mTALH) in HT (14.3 +/- 2.4 to 41.7 +/- 5.8 fm X 30 min-1 X mm-1, P less than 0.001) than in mTALH in C rats (14.4 +/- 2.8 to 110.1 +/- 24.9 fm X 30 min-1 X mm-1, P less than 0.001). In contrast, the adenylate cyclase response to AVP was not significantly different in collecting tubules of cortex, outer medulla, and inner medulla from C and HT rats, although a slight decrease in response to AVP was observed in cortical and outer medullary collecting tubules.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3037920 TI - Water permeability and particle aggregates in ADH-, cAMP-, and forskolin-treated toad bladder. AB - Osmotic water flow was used to evaluate total tissue water permeability (Ptissue), and luminal membrane particle aggregates, presumed sites for transmembrane water movement, were quantified to assess luminal membrane water permeability, in bladders treated with maximally stimulating concentrations of antidiuretic hormone (ADH), adenosine 3',5'-cyclic monophosphate (cAMP), and forskolin. Aggregates were as numerous and occupied the same fractional area of the luminal membrane in response to cAMP treatment (10 mM) as treatment with ADH (20 mU/ml). Ptissue in cAMP-treated tissues, however, was only half of that induced by ADH (P less than 0.001). A similar disparity in the relationship between aggregates and Ptissue occurred for additional bladders treated with 50 microM forskolin, which is known to increase endogenous cAMP to levels much greater than caused by maximally stimulating concentrations of ADH. Although Ptissue achieved with forskolin was the same in paired bladders treated with ADH, aggregates were far more numerous (P less than 0.05) and occupied much more membrane area (P less than 0.05) with forskolin. These observations are consistent with the view that aggregate appearance in the luminal membrane is a function of intracellular cAMP. The finding that the hydrosmotic response of toad bladder to both cAMP and forskolin compared with ADH stimulation is reduced relative to measured changes in luminal membrane aggregates suggests that the effect of ADH in altering water permeability involves additional regulation via a non-cAMP-mediated mechanism. This latter event would appear to be by an effect of ADH on the permeability of a resistance at a postluminal membrane site and/or possibly on the permeability of aggregates in the luminal membrane. PMID- 3037921 TI - Amiloride interacts with renal alpha- and beta-adrenergic receptors. AB - We have used radioligand binding techniques to assess whether amiloride and certain analogues of amiloride (ethylisopropyl amiloride and benzamil) can bind to adrenergic receptors in the kidney. We found that amiloride could compete for [3H]rauwolscine (alpha 2-adrenergic receptors), [3H]prazosin (alpha 1-adrenergic receptors), and [125I]iodocyanopindolol (beta-adrenergic receptors) binding in rat renal cortical membranes with inhibitor constants of 13.6 +/- 5.7, 24.4 +/- 7.4, and 83.6 +/- 13.5 microM, respectively. Ethylisopropyl amiloride and benzamil were from 2- to 25-fold more potent than amiloride in competing for radioligand binding sites in studies with these membranes. In addition, amiloride and the two analogues competed for [3H]prazosin sites on intact Madin-Darby canine kidney cells and amiloride blocked epinephrine-stimulated prostaglandin E2 production in these cells. We conclude that amiloride competes for binding to several classes of renal adrenergic receptors with a rank order of potency of alpha 2 greater than alpha 1 greater than beta. Binding to, and antagonism of, adrenergic receptors occurs at concentrations of amiloride that are lower than previously observed "nonspecific" interactions of this agent. PMID- 3037922 TI - Ionic dependence of active Na-K transport: "clamping" of cellular Na+ with monensin. AB - The Na+ ionophore monensin was used to study the Na+- and K+-dependence of ouabain-inhibitable 86Rb+ uptake in ARL 15 cells, a rat liver cell line. Graded concentrations of monensin rapidly induced incremental elevations of cellular Na+ that were stable for up to 2 h. In experiments in which cellular Na+ was thus "clamped" at various levels, the activation curve for ouabain-inhibitable 86Rb+ uptake as a function of intracellular Na+ was found to be steepest near basal Na+ levels (Hill coefficient approximately equal to 2.4), indicating that these cells can respond to relatively large changes in passive Na+ entry by increasing the race of Na-K pump function with only minimal increases in cellular Na+. Exposure of cells to monensin also permitted examination of the extracellular-K+ dependence of ouabain-inhibitable 86Rb+ uptake in the presence of saturating intracellular Na+ and yielded a Hill coefficient of approximately 1.5. The rate of ATP hydrolysis calculated from measurements of the maximal rate of ouabain inhibitable 86Rb+ uptake in intact cells was similar to the enzymatic Vmax of the Na+-K+-ATPase in cell lysates, suggesting that the Na+-K+-ATPase activity in these broken-cell preparations closely reflects the functional transport capacity of the Na-K pump. PMID- 3037923 TI - Effect of acute hypercapnia on PTH-stimulated phosphaturia in dietary Pi-deprived rat. AB - The effects of respiratory acidosis on renal inorganic phosphate (Pi) handling are controversial. Clearance experiments, therefore, were performed in fasted, chronically parathyroidectomized (PTX), dietary Pi-deprived rats. The objectives were twofold: to study the effects of compensated and uncompensated hypercapnia per se on renal Pi excretion and to examine the interaction between acute hypercapnia, dietary Pi, and parathyroid hormone (PTH) on the renal handling of Pi. Acute hypercapnia increased the plasma Pi (delta 2.82 +/- 0.65 mg/dl, P less than 0.05) without altering the glomerular filtration rate (GFR). The FEPi increased (delta 7.26 +/- 0.48%, P less than 0.001) but the TRPi/GFR also increased. PTH (3 U X kg-1 X h-1) superimposed on hypercapnia resulted in a plasma Pi comparable to hypercapnia alone. The FEPi (7.56 +/- 0.78 vs. 24.43 +/- 2.20%; P less than 0.001) was higher and the TRPi/GFR (117 +/- 4 vs. 80 +/- 2 micrograms/min, P less than 0.01) lower, in the former group. PTH infusion during normocapnia resulted in a lower FEPi (0.20 +/- 0.10 vs. 24.43 +/- 2.20%, P less than 0.001) and a higher TRPi/GFR (106 +/- 2 vs. 80 +/- 2 micrograms/min, P less than 0.01) compared with PTH infusion during hypercapnia. Urinary adenosine 3',5' cyclic monophosphate (cAMP) excretion was similar between the groups. During hypercapnia, when the extracellular acidemia was neutralized, the phosphaturic action of PTH persisted. These studies offer direct evidence that in chronically PTX, dietary Pi-deprived rats, the phosphaturic action of PTH is restored by hypercapnia per se. This effect appears to be independent of extracellular acidemia, changes in the plasma Pi and calcium, urinary pH and Na and cAMP excretion. PMID- 3037924 TI - Endogenous prostaglandins modulate autoregulation of renal blood flow in young rats. AB - After uninephrectomy in the young rat, renal blood flow (RBF) increases at normal renal perfusion pressure (RPP) but not at reduced RPP. To evaluate the role of endogenous prostaglandins on these effects, RBF was measured during reduction in RPP in rats that were uninephrectomized within 1 wk of age and studied at 33-41 days of age. After acute cyclooxygenase inhibition, autoregulation improved; RBF was unchanged at normal RPP but increased at lower pressures with concomitant decrease in renal venous plasma renin activity (PRA). Prostaglandin inhibition also improved autoregulatory efficiency in acutely uninephrectomized and sham operated young rats. Infusion of a thromboxane synthesis inhibitor had no effect on the pressure-flow relationship or on PRA, whereas angiotensin converting enzyme inhibition increased RBF at all measured RPP without affecting autoregulation. We conclude that increased RBF at normal RPP in the uninephrectomized young rat is not maintained by increased prostaglandin synthesis or decreased thromboxane- or angiotensin-dependent vasoconstriction. However, by maintaining increased vasoconstriction at reduced RPP, prostaglandin dependent renin release reduces autoregulatory efficiency in the young rat. PMID- 3037925 TI - Role of adenosine in regulation of cerebral blood flow: effects of theophylline during normoxia and hypoxia. AB - We studied the effects of the methylxanthine theophylline, an adenosine receptor blocker, on cerebral circulation. Cerebral blood flow (CBF) was measured by the retroglenoid outflow and microsphere techniques, and pial circulation changes were observed through a closed cranial window. Intraperitoneal administration of theophylline in normoxic animals resulted in a biphasic response of pial vessels and CBF. At low concentrations (0.05 mumol/g) of theophylline, pial vessel diameter and CBF decreased, whereas vasodilatation and hyperemia were observed at higher levels. After intraperitoneal administration of either 0.05 or 0.2 mumol/g, hypoxic hyperemia was attenuated both during short (c. 30 s) and sustained (c. 2-3 min) hypoxia, as was hypoxic pial arteriolar vasodilatation. These actions of theophylline appear to be due to adenosine receptor blockade, since micromolar concentrations were achieved in cerebrospinal fluid (CSF), and no increases in adenosine 3',5'-cyclic monophosphate concentrations in brain were noted. Moreover, theophylline (either intraperitoneal or topical) blocked pial vasodilatation caused by topically applied adenosine, but had little effect on hypercarbic hyperemia or pial vasodilatation induced by topically applied acetylcholine. The results of these studies suggest that adenosine is involved in the maintenance of resting cerebral vascular tone and has a paramount role in the regulation of CBF during hypoxia. PMID- 3037926 TI - Neurobiological mechanisms of panic anxiety: biochemical and behavioral correlates of yohimbine-induced panic attacks. AB - The effects of yohimbine, an alpha 2-adrenergic receptor antagonist, on anxiety, blood pressure, heart rate, and plasma levels of the norepinephrine metabolite 3 methoxy-4-hydroxyphenylglycol (MHPG) and cortisol were determined in 20 healthy subjects and 68 patients who had agoraphobia with panic attacks or panic disorder. Yohimbine produced panic attacks meeting DSM-III criteria in 37 patients and one healthy subject. The patients reporting yohimbine-induced panic attacks had significantly larger increases in plasma MHPG, cortisol, systolic blood pressure, and heart rate than the healthy subjects. These findings support the hypothesis relating high noradrenergic neuronal activity to the pathophysiology of panic attacks in a subgroup of panic disorder patients. PMID- 3037927 TI - ACTH response to corticotropin-releasing hormone. PMID- 3037928 TI - Alternate testing sites for AIDS virus. PMID- 3037929 TI - Effects on fertility of rabbits after active immunization with rat testicular cytochrome ct. AB - A Male rabbit was immunized with rat testicular cytochrome (Cyt) ct and mated with normal, unimmunized females. The matings resulted in abnormal pregnancies: no offspring or stillborn or undersized liveborn offspring weighing 25-30 gm each. Another unusual observation was that fur-pulling behavior, normally exhibited by pregnant female rabbits at the end of the gestational period, was absent in all of these pregnancies. Therefore, immunization of a normal rabbit with testicular cyt ct appeared to interfere with physiological and behavioral aspects of pregnancy in normal female rabbits. The immunological basis of these findings remains to be determined. PMID- 3037930 TI - Myofibroblastoma of the breast. Sixteen cases of a distinctive benign mesenchymal tumor. AB - The clinical and pathologic findings of 16 examples of a distinctive stromal tumor of the breast designated as "myofibroblastoma" are reported. Eleven of the 16 patients were men, and the average age at presentation was 63 years. Fourteen were treated by local excision and two by simple mastectomy. None of the lesions recurred or metastasized. The tumors were grossly nodular and well-demarcated from the surrounding mammary tissue. Ducts and lobules were not engulfed by the neoplasm. Microscopically, the lesions were formed by uniform, slender, bipolar spindle cells haphazardly arranged in fascicular clusters separated by broad bands of hyalinized collagen. Ultrastructural examination of four lesions identified a predominance of myofibroblasts. Immunoreactivity for S-100 protein and cytokeratin was absent in the 10 tumors examined, but desmin immunoreactivity was focally present in three lesions. The differential diagnosis of myofibroblastoma includes reactive processes and benign neoplasms such as nodular and proliferative fascititis, fibromatosis, spindle-cell lipoma, neurofibroma, neurilemmoma, and leiomyoma. Malignant neoplasms such as stromal sarcoma, malignant fibrous histiocytoma, and spindle-cell or metaplastic carcinoma should not be confused with a myofibroblastoma. The clinical significance of this entity lies primarily in its recognition as a distinctive benign neoplasm. PMID- 3037931 TI - Dracunculiasis eradication: a mid-decade status report. AB - A campaign to eradicate dracunculiasis has been underway from the beginning of the International Drinking Water Supply and Sanitation Decade (1981-1990), since providing safe drinking water is the most effective means to prevent that disease. About 120 million persons are estimated to be at risk of the infection in Africa, and 20 million more in India and Pakistan. Both major endemic countries in Asia have begun efforts to eliminate the disease, and by the end of 1986, national anti-dracunculiasis programs were underway or planned in 8 of the 19 affected African countries. In May 1986, the World Health Assembly adopted a resolution on the elimination of dracunculiasis-the first such resolution since the successful Smallpox Eradication Program. India, which began its Guinea Worm Eradication Program in 1980, has already eliminated the disease from one of seven endemic states, and reduced the total number of cases found through active surveillance by 35% between 1983 and 1985. In Cote d'Ivoire (Ivory Coast), the only African country to conduct active surveillance for dracunculiasis so far, an aggressive combined program of rural water supply, health education, and active surveillance has reduced the disease from 4,971 cases in 1976 to 592 cases in 1985. PMID- 3037932 TI - Sand fly fever-Naples infection in Egypt. AB - Two Egyptian male patients with sand fly fever-Naples virus infection are presented. The virus was isolated from one patient while both patients had diagnostic rises in indirect fluorescent antibody titers to the virus. The viral isolate, SFN 85055, grows to much higher titers and plaques more efficiently than the prototype sand fly fever-Naples virus and should facilitate work with this virus. PMID- 3037933 TI - Benzodiazepine receptor affinity alterations at physiologic temperature after chronic clonazepam exposure. AB - Cerebral cortical cell cultures obtained from fetal mice were exposed to 200 nM clonazepam (CZP) for 14 days and benzodiazepine (BDZ) receptor binding was measured on intact cells in situ at 37 degrees C. Total, specific, and CZP displaceable BDZ binding were significantly reduced from control values immediately after drug removal (77.7 +/- 1.4%, 75.1 +/- 3.0%, and 40.9 +/- 6.0% of control, respectively) but Ro5-4864-displaceable binding was not affected (87.6 +/- 5.1%). Binding returned to control values within 48 hours. Saturation analysis of the binding data indicated that a high-affinity binding site could not be detected in CZP-exposed cultures immediately after drug removal (162 nM versus 49 nM in controls, p less than .001), but was present 24 hours later. PMID- 3037934 TI - [Endocrine stress response in halothane, enflurane and isoflurane anesthesia in surgical interventions]. AB - The endocrine stress response under inhalation anesthesia with halothane, enflurane, and isoflurane was investigated in 30 patients during and after orthopedic surgery (Table 2). Plasma levels of adrenaline and noradrenaline (by HPLC/ECD), ADH, ACTH, and cortisol (by RIA), glucose, lactate, and free glycerol were determined before induction of anesthesia, 10 min after intubation, 10 min before the end of the operation, and 5 and 30 min after extubation. Statistical evaluation was undertaken by analysis of variance with repeated measures on one factor. P values of less than 0.05 were considered significant. There were no significant differences in the concentrations of plasma catecholamines (Table 4, Figs. 1 and 2), ADH, ACTH (Table 5, Figs. 3 and 4), or cortisol before and during surgery between the groups. ADH was lower in the halothane group 5 and 30 min after extubation (P = 0.05), which might be due to the prolonged elimination of halothane after anesthesia. Blood pressure, heart rate (Table 3), and plasma concentrations of glucose, lactate, and free glycerol (Table 6) were comparable in all groups. It is concluded that for clinical practice halothane, enflurane, and isoflurane are comparable in their influence on the surgical stress response. PMID- 3037935 TI - Transformation of mammalian cells with recombinant DNA directly from Seaplaque agarose. AB - Transfection of African green monkey kidney cells directly with recombinant DNA excised from, but still present in, Seaplaque agarose after electrophoresis, is described. Efficiencies of transfection increased by 30% when the gel was present compared with transfection in the absence of the agarose. Extraction of the DNA from the gel was not necessary, thereby obviating a purification step and the concomitant losses. To generate recombinant molecules bacterial plasmid sequences are not necessary, thereby reducing considerably the size of the recombinant molecule and removing extraneous and deleterious sequences, e.g., "poison sequences." Linear or circular DNA molecules could be transfected in the melted and diluted agarose with the same ease as in its absence. Hence linear partial ligation products can be excised from the gel after electrophoresis to generate recombinant DNA molecules directly in mammalian cells. PMID- 3037936 TI - Preparation of inositol phosphates from sodium phytate by enzymatic and nonenzymatic hydrolysis. AB - Procedures for preparing myo-inositol bis-, tris-, tetrakis-, and pentakisphosphates from sodium phytate were established. Hydrolysis was achieved by autoclaving or enzymatic treatment; the inositol phosphates were separated by anion-exchange chromatography and were identified by fast atom bombardment-mass spectrometry. Enzymatic hydrolysis was more specific than autoclaving for isomer formation, whereas autoclaving was more efficient for producing the bis- and trisphosphates, which did not accumulate in significant amounts under the conditions of enzymatic hydrolysis. Sodium salts of the inositol phosphates were more powdery and less hygroscopic than the potassium salts. The procedures were satisfactory for producing gram quantities of each inositol phosphate, amounts adequate for animal studies of effects on mineral bioavailability. PMID- 3037937 TI - Dideoxy sequencing of double-stranded DNA from poly(A)-extended 3'-ends. AB - A simple method has been developed for sequencing double-stranded DNA by the chain termination method. The DNA to be sequenced is cut with a restriction enzyme that leaves a 3'-overhang which is extended by terminal deoxynucleotidyltransferase with limiting amounts of dATP. The sequencing reaction is then primed with an oligo(dT) primer which has a base pair "anchor" complementary to the overhang generated by the restriction enzyme. The method presented here eliminates the need for subcloning of the DNA or sequencing by chemical modification. Furthermore, sequences of more than 300 nucleotides are obtained from any 3'-overhanging restriction end. PMID- 3037938 TI - Development of a specific radioimmunoassay for the placental folate receptor and related high-affinity folate binding proteins in human tissues. AB - High-affinity membrane-associated and soluble folate binding proteins (FBPs) from human placenta, milk, and KB cells appear to share antigenic determinants [A. C. Antony et al. (1981) J. Biol. Chem. 256, 9684-9692 and (1985) 260, 14911-14917]. Iodination of a highly purified preparation of placental folate receptor (PFR) by various techniques resulted in significant denaturation of the PFR as evidenced by additional peaks of radioactivity on Sephacryl S-200 gel filtration in 1% Triton X-100. These denatured species had similar molecular weights on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) as radioiodinated and native PFR, and were also recognized, albeit with less efficiency, by specific rabbit antiserum raised against purified PFR. Since these denatured species failed to bind folate, they were specifically excluded from 125I-PFR by their inability to bind pteroylglutamate-Sepharose. This ws accomplished in a single step by iodination of PFR bound to the affinity column and elution of 125I PFR under identical conditions that the native PFR was purified. The purified 125I-PFR comigrated with unlabeled PFR on SDS-PAGE and its elution profile on Sephacryl S-200 gel filtration was identical to radioligand bound PFR. The resulting radioimmunoassay standard curve using this affinity chromatography purified 125I-PFR, unlabeled PFR, and anti-human PFR serum had a range for measurement between 5 and 500 ng of PFR and was not affected by the concentration of folate in the sample. The practical utility of this radioimmunoassay for measuring cross-reacting material to the PFR was validated by its ability to quantitate the 40,000 and 160,000 Mr FBPs which are the two major forms of high affinity FBPs in human tissues. PMID- 3037939 TI - Solution-phase secondary-ion mass spectrometry of protonated amino acids. AB - Although sulfolane proved unexpectedly to be a poor solvent for solution-phase secondary-ion mass spectrometry of underivatized amino acids in the presence of thallium(I) salts, glycerol was somewhat more effective. Also, the addition of trifluoromethanesulfonic acid proved more effective than addition of the metal in generating molecular ion complexes. A convenient and reliable method for rapidly determining amino acid molecular ions is based on these observations. PMID- 3037940 TI - A continuous fluorescence assay for cyclic nucleotide phosphodiesterase hydrolysis of cyclic GMP. AB - The fluorescent 2'-methylanthraniloyl derivative of cyclic GMP undergoes a 45% decrease in fluorescence when it is cleaved by brain phosphodiesterase in the presence of calmodulin. This fluorescence decrease is dependent upon calcium, calmodulin, and phosphodiesterase, and correlates well (r = 0.996) with the disappearance of substrate as monitored by high-performance liquid chromatography. The Kd values determined by this fluorescence method and HPLC suggest that cyclic GMP and its fluorescent derivative exhibit similar kinetic parameters in their hydrolysis. PMID- 3037941 TI - Complete extraction in a form suitable for liquid scintillation counting of tritium-labeled proteins from polyacrylamide gels. AB - Large (200 mm3) slices of polyacrylamide gels crosslinked with N,N' diallyltartardiamide which contain tritium-labeled protein are readily solubilized in periodic acid for liquid scintillation counting of radioactivity, but the apparent recovery of label never exceeds 82%. Extraction of the slices with two commercial solubilizers at 60 degrees C gave recoveries of 82-90% which were not improved by prolonged incubation. Treatment of the slices at ambient temperature with 1.0 ml of 2% sodium periodate for 30 min followed by the addition of 0.7 ml of aqueous tetrabutylammonium hydroxide (40% w/v) gives solutions which can be immediately counted at 35% efficiency with low background and with 100% recovery of tritiated protein PMID- 3037942 TI - Rapid resolution of phenylthiohydantoin amino acids by thin-layer chromatography on silica gel plates impregnated with transition metal ions. AB - The resolution of a 15-component mixture of phenylthiohydantoin (PTH) amino acids using metal ion impregnated silica gel plates is reported. The spots are located by exposing the chromatograms to an iodine chamber. The method provides a rapid, simple, and less expensive chromatographic system, provides resolution for certain difficult combinations, and leaves the PTH amino acids unaltered chemically. PMID- 3037943 TI - Use of Fourier transform 1H NMR in the identification of vitamin D2 metabolites. AB - The use of Fourier transform 1H NMR to characterize vitamin D2 metabolites is described. A 300-MHz spectrometer capable of generating a 6-microseconds pulse and a sweep width of 4000 Hz was used. High-resolution spectra were obtained on 5 micrograms of material using standard 5-mm NMR tubes fitted with glass inserts and isotopically enriched chloroform-d solvent. The data acquisition time under these conditions was 4 h. Application of this technique to a variety of both synthetic and naturally occurring vitamin D2 metabolites, in addition to synthetic delta 22-1,25-dihydroxyvitamin D3, resulted in the reassignment of the chemical shifts for the C-21 and C-28 methyl groups of vitamin D2. The C-21 methyl group resonance is now assigned to the doublet appearing at delta 1.01, whereas the C-28 signal corresponds to the doublet at delta 0.90. An examination of the spectrum of 24 (R),25-dihydroxyvitamin D2 also led to the reassignment of the side-chain methyl group resonances. This technique is an additional means of identifying microgram quantities of vitamin D metabolites. PMID- 3037944 TI - Rapid isolation of eukaryotic DNA. AB - Guanidine hydrochloride (GHCl) is frequently used for the isolation of RNA from eukaryotic cells. However, it appears that its use for the rapid isolation of chromosomal DNA has been little appreciated. Intact and readily digestible DNA was prepared from a range of murine tissues and cell lines which had been dispersed in GHCl and ethanol precipitated, and then rinsed in 70% ethanol and resuspended in 10 mM Tris-HCl, 1 mM EDTA, pH 7.5. This simple technique is ideally suited to the preparation of DNA from large numbers of tissue samples. PMID- 3037945 TI - An optimized thymidylate kinase assay, based on enzymatically synthesized 5 [125I]iododeoxyuridine monophosphate and its application to an immunological study of herpes simplex virus thymidine-thymidylate kinases. AB - The biological synthesis and purification of 5-[125I]iododeoxyuridine monophosphate (IdUMP) are described. The specificity of IdUMP as substrate in the thymidylate monophosphate kinase (TMPK) assay is demonstrated, and a 100-fold gain in sensitivity as compared to the conventional TMPK assay is shown. TMPK measurements of isozymes derived from herpes simplex virus (HSV)-infected cells, uninfected cells, and tumor biopsies were performed. The results showed a significant difference in dependence of phosphate donor concentration present for TMPK activity from HSV-infected cells compared to the corresponding activity from uninfected cells, while only a minor difference in pH optima was observed for these enzyme activities. The increased sensitivity made it possible to detect and quantify HSV TMPK-blocking antibodies (ab) present in human sera. Sera from HSV ab-positive individuals were found to block the two HSV TMPKs to varying degrees and with different specificities. The immunological relationship between the TMPK and thymidine kinase (TK) induced by HSV-1 and HSV-2, respectively, was studied by comparing the capacities of different sera to block the two enzymatic activities. The results showed that the capacity to block HSV-1 TK and TMPK was proportional for all of the sera studied, while sera that preferentially blocked only the HSV-2 TMPK or HSV-2 TK were found. It was concluded that the HSV-2 TMPK and TK activities are less related than the corresponding activities for HSV-1 and that the HSV-2 enzyme activities are mediated by different catalytic sites. PMID- 3037946 TI - Dot assay for neomycin phosphotransferase activity in crude cell extracts. AB - A dot assay for determining neomycin phosphotransferase (NPT II) activity in crude cell extracts has been developed. The assay provides for the rapid screening of large numbers of cell cultures generated in gene transformation experiments using NPT II as a dominant selectable marker. Currently, the commonly used procedure for NPT II assay employs a time-consuming electrophoretic protein separation step to eliminate a positive interference resulting from putative protein kinase activities present in crude cell extracts. The dot method we have developed is based upon the ability of nitrocellulose membrane to eliminate that positive interference without a prior protein separation step. It provides a sensitive, reproducible, and significantly more convenient and rapid means of screening large numbers of cell extracts in order to distinguish cultures producing high levels of NPT II from those that do not. PMID- 3037947 TI - Regional anesthesia in a child with epidermolysis bullosa. PMID- 3037948 TI - Sodium bicarbonate buffers gastric acid during surgery in obstetric and gynecologic patients. PMID- 3037949 TI - Acid-base and electrolyte balance in dairy heifers fed forage and concentrate rations: effects of sodium bicarbonate. AB - Eight Holstein heifers were fed diets of alfalfa hay, corn silage, or finely ground corn grain with or without NaHCO3 in a rotating experimental design. Acid base status and renal excretion of electrolytes were evaluated during short-term (1 and 5 day) and long-term (24 day) feeding trials. Heifers fed alfalfa hay had a greater metabolic buffering capacity than did heifers fed corn silage. Heifers fed grain had lower blood pH and bicarbonate values than did those fed the forage diets. The most pronounced effects of grain-feeding were aciduria and phosphaturia. Aciduria did not occur when NaHCO3 was added to the grain at 2% of the ration on a dry matter basis. Grain-fed heifers had significantly (P less than 0.05) lower blood and urine pH and bicarbonate values, and excreted significantly more calcium, phosphorus, and magnesium in the urine than did those fed grain plus NaHCO3. Sodium bicarbonate, as a 2% dietary supplement, counteracted many effects of high-grain diets. PMID- 3037950 TI - In vitro viral expression as a criterion for development of control procedures for enzootic bovine leukosis. AB - In a university beef herd of 304 cattle in which six died of lymphosarcoma between 1980 and 1984, 77% of the Angus and 26% of the Charolais cattle were determined to be infected with bovine leukemia virus (BLV). Changes in iatrogenic procedures were initiated as early control measures. In vitro viral expression (VE) was used as a criterion to identify cattle for subsequent segregation or culling. This involved determinations of percentages of BLV-associated lymphocyte profiles among thin-sectioned Ficoll-Paque-isolated blood lymphocytes that were processed into plastic after culture for 48 h. Cattle retained until completion of nutritional studies or as breeding stock were separated into two groups. The BLV-seronegative cattle, BLV-seropositive cattle with 0% VE, and BLV-seropositive cattle with 1% to 4% VE were placed in group 1. Seropositive cattle with greater than or equal to 5% VE were placed in group 2. In 1985, evaluation of in vitro VE in 108 mature BLV-seropositive cattle retained for breeding revealed 36 (33%) had no observable VE. In 1986, 58 of 108 cattle were available to be reexamined, and 21 (36%) had 0% VE in both years. The VE expression values for individual cattle were generally comparable over the 2-year period. Of 48 initial seronegative breeding stock housed in group 1 with BLV-seropositive cattle with low or no VE, 21 (44%) seroconverted during 1985 to 1986. A positive correlation of 0.585 was found between VE and age-related absolute lymphocyte number. PMID- 3037951 TI - Frequency and significance of feline leukemia virus infection in necropsied cats. AB - Feline leukemia virus (FeLV) infection was diagnosed immunohistologically on paraffin-embedded tissues obtained from 1,095 necropsied cats. Significant association of FeLV infection was demonstrated by chi 2 and Fisher's tests with various conditions and diseases (ie, anemia, tumors of the leukemia/lymphoma complex, feline infectious peritonitis, bacterial infections, emaciation, FeLV associated enteritis, lymphatic hyperplasia, and hemorrhage). Unexpected findings associated with FeLV infection were icterus, several types of hepatitis, and liver degeneration. A negative association with FeLV infection was found for most parasitic and viral infections, including feline panleukopenia. Neither positive nor negative associations were established for FeLV infection and most forms of nephritis, including severe glomerulonephritis. Feline leukemia virus-infected cats were significantly (Kruskal-Wallis test) older than were FeLV-negative cats with the same nonneoplastic FeLV-associated diseases. PMID- 3037952 TI - Pathogenesis of rotavirus infection in various age groups of chickens and turkeys: clinical signs and virology. AB - Age-related susceptibility patterns of turkeys, broilers, and specific pathogen free (SPF) White Leghorn chickens to experimentally induced infection with turkey or chicken rotavirus isolates were compared. The following determinants were evaluated: clinical signs, onset and duration of virus production, viral titers, involvement of intestinal villi in the replication of the virus, and the development of antibodies against the virus. Older turkeys and chickens were more susceptible than were their younger counterparts, turkeys were more susceptible than were broiler and White Leghorn chickens (regardless of age), and broiler chickens were slightly more susceptible than were age-matched White Leghorn chickens. Turkeys developed diarrhea, accompanied by high viral titers within 1 day after inoculation with virus. Viral antigen was found in the epithelial cells of the intestinal villi throughout the intestinal tract and some cells of the cecal tonsils. Antibodies could be detected as early as 4 to 5 days after inoculation. These findings were more pronounced in turkeys inoculated at 112 days of age than in birds inoculated at a younger age. Age-related susceptibility patterns were similar in White Leghorn and broiler chickens. Infection was subclinical in birds less than 56 days old, whereas older birds developed soft feces. Egg production in the White Leghorn chickens decreased after being inoculated with virus at 350 days of age. PMID- 3037953 TI - Detection of bovine herpesvirus-1 nucleic acid sequences, using a dot-blot hybridization procedure. AB - A sensitive and specific procedure for the detection of Argentine isolates of bovine herpesvirus-1 was developed. The procedure was based on a dot-blot, nucleic acid hybridization, using 32P, nick-translated, plasmidic probes. The probes contained cloned Bam H1 restriction fragments in the left half of the viral genome. The detection limit of the procedure was 10 pg of viral DNA. PMID- 3037954 TI - Localization of quantitative changes in pulmonary beta-receptors in ovalbumin sensitized guinea pigs. AB - Impaired beta-receptor function has been postulated as one factor contributing to airway hyperreactivity in asthmatic patients. Although numerous indirect studies have cast doubt on this theory, none of these previous investigations has been able to directly measure changes in beta-receptor number on intrapulmonary structures capable of affecting the physiologic changes seen in this disease state. To help clarify the intrapulmonary location of such changes, a model of allergic bronchoconstriction was prepared by sensitizing guinea pigs to ovalbumin intraperitoneally (ip) 2 wk prior to testing (Group S). A second group of animals was sensitized to ovalbumin, then 2 wk later partially desensitized (Group D) during a 4- to 6-wk period by repeated exposure to increasing doses of nebulized ovalbumin with epinephrine rescue. Control animals received ip administered and nebulized normal saline alone. Pulmonary function assessed by plethysmography revealed an increase in airway resistance to 294 +/- 42% (SE) of control in Group S (p less than 0.005) and a decrease in dynamic compliance to 76 +/- 8% of control in Group D and 39 +/- 10% of control in Group S (p less than 0.002) after exposure to nebulized ovalbumin. Using L-[3H] dihydroalprenolol ([3H] DHA), beta receptors were autoradiographically localized and quantitated in lung sections from all 3 groups. Significant decreases (p less than 0.02) in 3H-DHA binding were noted in alveolar and conducting airway epithelium, and bronchiolar and vascular smooth muscle in ovalbumin-exposed animals.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3037955 TI - Leukotrienes and the pulmonary microcirculation. PMID- 3037956 TI - Cyclic AMP-dependent regulation of lipid mediators in white cells. A unifying concept for explaining the efficacy of theophylline in asthma. AB - Activation of white cells, including the neutrophil, eosinophil, basophil, and mast cell, has long been known to be suppressed by high, nonphysiological levels of E-prostaglandins (PGE). In contrast, PGE at levels consistent with an interaction with the PGE receptor (5 X 10(-9) M) have recently been shown to suppress leukotriene (LT) and prostaglandin (PG) production by neutrophils and eosinophils. This occurs by cyclic AMP-dependent inhibition of release of substrate arachidonic acid (AA) from phospholipid pools. The additional observation that indomethacin (10(-9) M) enhances release of eicosanoids by suppressing endogenous PGE2 acting to increase cAMP levels in these cells. Theophylline and other phosphodiesterase inhibitors precisely duplicate the action of PGE2, and the combined effects of such phosphodiesterase inhibitors and adenylate cyclase stimulators are synergistic. The mechanism of action of theophylline in asthma is not know, although it is generally agreed that its effect is a direct one on the bronchial smooth muscle. The findings described in this report now permit the bronchial smooth muscle, but is primarily one of suppressing mediator release from relevant white cells by inhibition of cAMP phosphodiesterase, an action that is enhanced by the presence of inflammatory prostaglandins in the lung. PMID- 3037958 TI - Mucosal proctectomy and ileoanal pull-through technique and functional results in 23 consecutive patients. AB - Mucosal proctectomy with ileoanal pull-through in the treatment of ulcerative colitis and familial polyposis provides a technique for the preservation of the anal sphincters and relatively normal mechanisms of continence. Five patients had straight ileoanal anastomosis while 18 had the construction of a J-pouch. A two team approach was used for simultaneous abdominal and perineal procedures to facilitate a shortened operating time. A loop ileostomy was routinely used in the postoperative period and was closed an average of 4.5 months (range: 2-16 months) later without complication. Prolonged preoperative hospitalization was rarely necessary and outpatient steroid enema preparation was routinely used. There were no deaths. Nineteen patients with functioning pull-through procedures have been followed an average of 23 months (range: 3-42 months). Two other patients have not had ileostomy closure because of complications. The two remaining patients had intractable diarrhea and have since undergone conversion to a permanent ileostomy. The 19 patients are continent, having three to nine bowel movements each day. Nearly all wear a perineal sanitary pad because of rare, unpredictable leakage of small amounts of fluid, especially at night. Complications were significant in this group of patients. Intestinal obstruction was a frequent problem, occurring in 52 per cent of the entire series and necessitating reoperation in 22 per cent. Anal stricture was a problem in another five patients. A variety of other minor problems occurred and most were treated nonoperatively. In spite of moderate diarrhea and occasional leakage of stool, all patients with functioning pull-through procedures prefer their current status to life with an ileostomy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3037957 TI - Bronchoalveolar neutrophilia during late asthmatic reactions induced by toluene diisocyanate. AB - The mechanism by which late asthmatic reactions are induced by toluene diisocyanate (TDI), a low molecular weight chemical that causes occupational asthma in exposed subjects, is unknown. We investigated whether early and late asthmatic reactions induced by TDI are associated with changes in airway responsiveness to methacholine and airway inflammation as determined by bronchoalveolar lavage. We measured FEV1 before and at regular intervals after exposure to TDI, and performed dose-response curves to methacholine and bronchoalveolar lavage at 8 h after TDI in a group of 6 subjects with late asthmatic reactions and in 6 subjects with only early asthmatic reactions. The same procedure was followed 2 h after TDI in a group of 6 subjects with previously documented late asthmatic reactions and in a group of 6 subjects without any previously documented asthmatic reaction after TDI. In subjects with late asthmatic reactions, neutrophils were increased at both 2 and 8 h, and eosinophils and airway responsiveness were increased only at 8 h. By contrast, neutrophils, eosinophils and airway responsiveness were not increased at 8 h after TDI in subjects with an early asthmatic reaction or at 2 h after TDI in normal control subjects. These results suggest that late asthmatic reactions to TDI, and the associated increase in airway responsiveness, may be caused by airway inflammation. PMID- 3037959 TI - Routine use of the thallium-technetium scan prior to parathyroidectomy. AB - The merit of the preoperative localization of parathyroid tumors with thallium technetium subtraction imaging is a subject of current debate in the treatment of primary hyperparathyroidism. Eighty patients with hyperparathyroidism underwent preoperative subtraction scintigraphy with 201Tl Cl and 99mTcO4; scan results were correlated with the operative identification and histopathology of the resected parathyroid tissue. The true-positive, false-positive and false-negative rates of these scans were compared between patients with tumors in normal and ectopic anatomic locations and between patients undergoing an initial and reoperative neck exploration. The scan was clearly valuable in patients with one or more prior neck explorations. True-positive scans were obtained in seven (77%) of nine such patients. The scan was also valuable in patients who had ectopic tumors (six mediastinal, seven intrathyroid, and two in the carotid sheath). Twelve (80%) of these 15 ectopic tumors were correctly localized either prior to their first operation or before subsequent explorations. In contrast, only 33 (50%) of 65 patients had a true-positive scan prior to their first operation and when the tumor was not in an ectopic location. In summary, in this series, the thallium-technetium scan was correct in only 50 per cent of patients undergoing an initial operation. However, it was positive in 77 per cent of patients who had at least one prior neck exploration and in 80 per cent of patients with an ectopic parathyroid tumor. These results support the selective use of this valuable imaging and localization tool. PMID- 3037960 TI - 9-(1,3-Dihydroxy-2-propoxymethyl)guanine (ganciclovir) in the treatment of cytomegalovirus gastrointestinal disease with the acquired immunodeficiency syndrome. AB - 9-(1,3-dihydroxy-2-propoxymethyl) guanine (ganciclovir) was used to treat 41 patients (median age, 37 years) with the acquired immunodeficiency syndrome and cytomegalovirus gastrointestinal infection. Sites of infection were the colon in 31, the esophagus in 5, the rectum in 4, and the small bowel in 1. Patients received ganciclovir, 5 mg/kg body weight, intravenously every 12 hours for 14 days. Clinical improvement was seen in 30 patients and virologic response in 32. Mainly hematologic toxicity occurred: moderate leukopenia (1000 to 1900/mm3) was seen in 7 patients and severe (less than 1000/mm3) in 1, and moderate neutropenia (500 to 1000/mm3) in 5 and severe (less than 500/mm3) in 1. A cutaneous rash developed in 2 patients. Median overall survival was 16 weeks (range, 2 to 56). Cytomegalovirus recurred in 13 patients; median time to recurrence was 9 weeks from the start of treatment. Ganciclovir may be effective in treating cytomegalovirus gastrointestinal disease in patients with the acquired immunodeficiency syndrome. PMID- 3037961 TI - Options for isolation precautions. PMID- 3037962 TI - Effects of ethanol on proteins of mitochondrial membranes. PMID- 3037963 TI - Effects of ethanol on calcium homeostasis in rat hepatocytes and its interaction with the phosphoinositide-dependent pathway of signal transduction. PMID- 3037964 TI - Microsomal generation of hydroxyl radicals: its role in microsomal ethanol oxidizing system (MEOS) activity and requirement for iron. PMID- 3037965 TI - Adaptation to ethanol in cultured neural cells and human lymphocytes from alcoholics. PMID- 3037966 TI - Third Colloquium in Biological Sciences: Cellular signal transduction. PMID- 3037967 TI - Differential transmembrane signaling in B lymphocyte activation. PMID- 3037968 TI - Interactions between grafted serotonin neurons and adult host rat hippocampus. AB - We have followed the development of physiological and functional properties of serotonin-containing raphe neurons grafted into an adult host hippocampus as a model system for graft-host interactions. These raphe cells have no clear identifying properties on the day of grafting: they develop them while growing in the host. Raphe neurons, recorded 1 month after grafting, possess adult normal physiological properties. These include high input resistance, slow membrane time constant, lack of inward rectification, a transient outward rectification, broad spikes having a Ca2+ component, lack of accommodation, and a large afterhyperpolarization. The graft is first innervated by host fibers and later projects to the host tissue. When stimulated, postsynaptic hyperpolarized responses are recorded in hippocampal neurons. In the freely moving rat, raphe grafts can restore sleep-wakefulness variations in an evoked population response of the hippocampus to afferent stimulation, which is eliminated by depletion of serotonin. These studies illustrate that grafted serotonin neurons develop functional relations with a host brain. PMID- 3037969 TI - Morphological and behavioral characteristics of embryonic brain tissue transplants in adults, brain-damaged subjects. PMID- 3037970 TI - Receptor characteristics and behavioral consequences of kainic acid lesions and fetal transplants of the striatum. PMID- 3037971 TI - Interaction of met-enkephalin with human granulocytes. AB - Met-enkephalin has been found to have an effect on cell shape and motility of human polymorphonuclear leukocytes (PMNs). Specific binding of tritium-labeled Met-enkephalin to the cells could not be demonstrated. There was a rapid proteolytic degradation of the peptide in the medium, followed by uptake of the labeled tyrosine. The peptidase recognizes the N-terminal sequence of various endogenous opioid peptides. The protease inhibitors bestatin and bacitracin had but little effect on the degradation of Met-Enk. PMID- 3037972 TI - Possible correction of defective polymorphonuclear cell functions in type-2 diabetes mellitus by met-enkephalin. AB - In vitro pretreatment with Met-Enk of human PMNLs obtained from patients suffering from type-2 diabetes mellitus resulted in the normalization of the ROS generating system. It is assumed that Met-Enk has a modulating effect on the arachidonic acid metabolism, with a consecutive increase of the LTB4 release. PMID- 3037973 TI - Neuropeptides modulating macrophage function. AB - The immune system and the neuroendocrine system affect each other via molecules and receptors shared by both systems. Neuroendocrine hormones may act either positively or negatively in regulating the activities of a key cell of the immune system, the macrophage. For example, adenocorticotropic hormone (ACTH), somatostatin, and substance P are all capable of increasing the cytotoxicity of macrophages against tumor cells. However, ACTH and somatostatin, but not substance P, can also block the tumoricidal activity of macrophages induced by recombinant gamma interferon (IFN-gamma), a non-neuroendocrine immunomodulating hormone. In contrast, substance P increased tumoricidal activity, both independent of IFN-gamma and in addition to IFN-gamma. Neurotensin, alpha endorphin, beta-endorphin, met-enkephalin, vasopressin, and substance K did not affect tumoricidal function, either alone or in combination with IFN-gamma. Substance P, but not the other neuropeptides, increased substantially the proportion of macrophages able to secrete superoxide ions, suggesting a possible influence on macrophage capacity to deal with microbial infection. Such positive and negative modulation of macrophage effector functions could contribute to the influence of cognitive stimuli in infection and neoplasia. PMID- 3037974 TI - Developmental expression and cellular localization of the beta-adrenergic receptor in the murine thymus. PMID- 3037975 TI - Consequences of bilateral brain neocortical ablation on imuthiol-induced immunostimulation in mice. PMID- 3037976 TI - Anterior hypothalamic lesions influence respiratory airway hyperreactivity. PMID- 3037977 TI - Regulation of natural immunity (NK activity) by conditioning. AB - A paradigm is described for the conditioning of the NK response. Mice were exposed to a classical conditioning procedure in which saccharin and lithium chloride (LiCl) served as the conditioned stimulus (CS) paired with an injection of either poly I:C or cytoxan, the unconditioned stimulus (US). Poly I:C was used as the US for enhancement and cytoxan served as the US for suppression. Subsequent exposure to the CS either enhanced or suppressed the NK activity in the conditioned animals. Our studies showed that only one association was needed for the learning to take place, training to consumption of water was not necessary, and the conditioned (enhanced) response could be recalled as soon as 7 days after pairing of the US and CS. PMID- 3037978 TI - Effects of phosphatidylserine on immunologic indices in aged patients. PMID- 3037979 TI - Interleukin-1 and glucocorticoid hormones integrate an immunoregulatory feedback circuit. PMID- 3037980 TI - Centrally active peptides. Are they useful agents in the treatment of obesity? PMID- 3037981 TI - Structural and functional responses of enzymes to immobilization: new insights from EPR spectroscopy. PMID- 3037982 TI - [The EEC syndrome (ectrodactyly, ectodermal dysplasia, cleft lip and palate)]. PMID- 3037983 TI - Further application of VM-plasty. AB - The use of VM-plasty in the repair of web contracture and its application for other regions are presented. We refined VM-plasty so that both ends remain in the "dog ear" form, eschewing the conventional excision and instead making flaps that are set in a zigzag shape. This procedure results in an increase in the effect of extension to the lateral direction, in addition to preventing recontracture. This method is used not only to repair finger web contracture but also for other contracted scars. No recontracture has been observed, and good results have been obtained. PMID- 3037984 TI - Cystosarcoma phylloides treated by excision and immediate reconstruction with silicon implant. AB - A case of cystosarcoma phylloides in an 11-year-old girl is described. Excision was followed immediately by reconstruction with a silicon implant. To the best of our knowledge, this is the first report of this technique in an adolescent girl. The etiology, incidence, diagnosis, and treatment of cystosarcoma phylloides are described. PMID- 3037985 TI - In vitro antimicrobial activity of diethyldithiocarbamate and dimethyldithiocarbamate against methicillin-resistant Staphylococcus. AB - Staphylococcus aureus has appeared which is highly resistant to both methicillin and aminoglycosides. Current therapy involves long-term intravenous therapy of vancomycin. Since vancomycin is currently the only drug used to treat these patients, there is a need to develop additional antimicrobial therapy. The in vitro antimicrobial effect of the metal chelator, diethyldithiocarbamate (DDTC) and its structural analog dimethyldithiocarbamate (DMTC) were investigated. Both DDTC and DMTC were effective against S. aureus including methicillin-resistant S. aureus (MRSA). By agar diffusion, DDTC at 10 micrograms per disk produced zone sizes of 12 to 21 mm and at 100 micrograms per disk produced zone sizes of 26 to 34 mm against MRSA. The DMTC produced slightly greater zone sizes against MRSA of 16 to 24 mm and 24 to 37 mm for 10 micrograms per disk and 100 micrograms per disk, respectively. The minimum inhibitory concentration (MIC) for DMTC against MRSA was 6 micrograms per ml. Both DDTC and DMTC were also effective against enterococci, Proteus mirabilis, Klebsiella pneumoniae, Klebsiella oxytoca, Escherichia coli, Enterobacter cloacae, Enterobacter aerogenes, Salmonella species, Serratia marcescens and Citrobacter freundii at 100 micrograms per disk. The MICs of DMTC for Klebsiella pneumoniae, Klebsiella oxytoca, Escherichia coli, Salmonella and Citrobacter freundii were approximately 128 micrograms per ml while the MICs for Proteus vulgaris, Proteus mirabilis, Pseudomonas aeruginosa and Serratia marcescens was greater than or equal to 256 micrograms per ml. In addition, DMTC was synergistic with gentamicin against MRSA and coagulase negative staphylococcus species, Enterobacter cloacae, Klebsiella pneumoniae and Pseudomonas aeruginosa. Additive and synergistic effects of DMTC were displayed with gentamicin against S. aureus including methicillin-resistant S. aureus.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3037986 TI - Beyond receptors: multiple second-messenger systems in brain. AB - Second-messenger systems play a major role in mediating neurotransmitter actions. In recent years our understanding of the organization and function of two prominent second-messenger systems has progressed rapidly--the adenylate cyclase and phosphoinositide systems. Guanosine triphosphate-binding proteins, which are especially abundant in brain, couple transmitter receptors to the key second messenger generating enzymes in both of these systems. Whereas activation of adenylate cyclase produces a single intracellular messenger, cyclic AMP, stimulation of the phosphoinositide system generates at least two, inositol trisphosphate and diacylglycerol. Inositol trisphosphate mobilizes calcium from intracellular stores, and diacylglycerol, like cyclic adenosine monophosphate, activates a phosphorylating enzyme, protein kinase C. These second-messenger systems are particularly enriched in the brain where they modulate many aspects of synaptic transmission. PMID- 3037987 TI - Adrenoleukodystrophy: dietary oleic acid lowers hexacosanoate levels. AB - Adrenoleukodystrophy (ALD) is an X-linked disorder characterized by demyelination, adrenal insufficiency, and accumulation of saturated very-long chain fatty acids (VLFA), particularly hexacosanoate (C26:0). We treated 5 patients with adrenoleukodystrophy (3 males and 2 symptomatic female carriers) for 6 months with a diet enriched in oleic acid (C18:1) and moderately restricted in C26:0. Elevated plasma and erythrocyte levels of C26:0 decreased in a time dependent manner during treatment. Total plasma C26:0 concentration was lowered by 50 +/- 9% (p less than 0.01); it became normal in the female carriers. The total erythrocyte level of C26:0 decreased (44 +/- 5%; p less than 0.001) into the normal range in all patients. Significant decreases were noted in the saturated VLFA composition of plasma and erythrocyte sphingomyelin and erythrocyte phosphatidylcholine during dietary treatment. In general, decreases in saturated VLFA levels were accompanied by increases in monounsaturated VLFA levels, while total VLFA values did not change. This novel approach to the treatment of adrenoleukodystrophy, in which there is an exchange of monounsaturated VLFA for the more toxic saturated VLFA, may prove clinically beneficial in this disorder. PMID- 3037988 TI - Markers of focal epilepsy. PMID- 3037989 TI - Protective effects of liposome-entrapped superoxide dismutase on posttraumatic brain edema. AB - Oxygen-derived free radicals and membrane lipid peroxidation have been postulated to be involved in brain edema and cell death, secondary to ischemia and traumatic injury. Using a model of brain edema induced by cold-induced injury, we have demonstrated an early elevation of superoxide radicals followed by permeability changes in the blood-brain barrier and development of edema in injured brain. Intravenous injection of liposome-entrapped copper-zinc-superoxide dismutase 5 minutes before the injury-enhanced entry of the enzyme into endothelial cells of the blood-brain barrier of injured brain reduced the brain level of superoxide radicals and ameliorated blood-brain barrier permeability changes and brain edema. Identical treatment 5 minutes after injury was also effective. These data demonstrate that superoxide radicals play an important role in the delayed development of vasogenic brain edema following brain injury. PMID- 3037990 TI - Progressive cytochrome c oxidase deficiency in a case of Kearns-Sayre syndrome: morphological, immunological, and biochemical studies in muscle biopsies and autopsy tissues. AB - We report biochemical, immunological, and morphological findings in a patient with fatal Kearns-Sayre syndrome. Histochemical and biochemical findings from muscle biopsy specimens obtained 7 years apart documented the disease's evolution from a mild mitochondrial disorder affecting a small proportion of muscle fibers to a severe disorder affecting a large proportion of muscle fibers. Cytochrome c oxidase activity in muscle declined profoundly as the disease progressed, although the level of enzyme protein was normal, as shown by immunochemical techniques. Other organs were severely affected by the disease. Examination of postmortem tissue showed spongiosis in the frontal cortex, diffuse loss of Purkinje cells in the cerebellum, liver steatosis, and heart fibrosis with mitochondrial abnormalities. Cytochrome c oxidase activity was only slightly reduced in these organs. PMID- 3037992 TI - Biochemical and genetic characterization of type I familial amyloidotic polyneuropathy. AB - Type I familial amyloidotic polyneuropathy is an autosomal dominant, inherited systemic amyloidosis characterized initially by dissociated sensory disturbance and autonomic dysfunction. The amyloid fibril protein seen in patients of Portuguese, Japanese, and Swedish descent in the U.S. mainly consists of a variant form of transthyretin (also called prealbumin) with the substitution of methionine for valine at position 30. Methods have been developed to detect this variant transthyretin in the serum, and to detect a base change in a mutated transthyretin gene. The biochemical and genetic abnormalities in transthyretin are completely linked to the clinical diagnosis of type I familial amyloidotic polyneuropathy. These diagnostic methods may allow early diagnosis and genetic counseling to avoid transmission of this intractable disorder to the next generation. PMID- 3037991 TI - Beta-endorphin, ACTH, and catecholamine responses in chronic autonomic failure. AB - Plasma epinephrine (EPI), norepinephrine (NE), beta-endorphin, and corticotropin (ACTH) responses were measured during insulin-induced hypoglycemia in normal subjects and in patients with either multiple system atrophy (MSA) or idiopathic orthostatic hypotension (IOH). In normal subjects, there was a striking rise in EPI, NE, beta-endorphin, and ACTH following the nadir of hypoglycemia. Both beta endorphin and ACTH responses were significantly lower than normal in patients with MSA, in contrast to normal levels in IOH patients. No correlation was observed between the degree of adrenergic insufficiency and the beta-endorphin and ACTH responses. The normal peptide responses in IOH are consistent with involvement limited to the peripheral sympathetic nervous system, whereas lesions in the central nervous system in MSA interfere with release of beta-endorphin and ACTH in response to hypoglycemia. The strong correlation between beta-endorphin and ACTH levels is consistent with their common origin. Peripheral adrenergic activity is not essential for beta-endorphin and ACTH release in humans. PMID- 3037993 TI - Axilla skin biopsy: a reliable test for the diagnosis of Lafora's disease. AB - Skin biopsies from the axilla were examined in 7 patients with clinical signs and symptoms of Lafora's disease, in 9 relatives (7 parents and 2 siblings), and in 17 patients with chronic or progressive degenerative neurological disorders and chronic epilepsy, who served as controls. In the biopsies of all patients with Lafora's disease, typical periodic acid-Schiff (PAS)-positive inclusions were present in the myoepithelial cells of the secretory acini of the apocrine glands and/or in the cells of the eccrine duct. No abnormalities were found in the biopsies from relatives and controls. We conclude that axilla skin biopsy is an easy and reliable test for diagnosing Lafora's disease, and is preferable to liver biopsy, but has no value for carrier detection. PMID- 3037994 TI - Diencephalic seizures: responsiveness to bromocriptine and morphine. AB - Two patients with posttraumatic diencephalic seizures, characterized by autonomic dysfunction and extensor posturing, had partial responses to bromocriptine and complete responses to morphine. Probable synergism between the two agents was noted. These 2 cases suggest the potential effectiveness of this regimen for the treatment of diencephalic seizures, raise questions regarding the role of the dopaminergic and opioid systems in this disease entity, and support the hypothesis that diencephalic seizures represent a release phenomenon in the brain. PMID- 3037995 TI - [Genetic determinants of resistance to antibiotics in Pseudomonas bacteria]. AB - Seven hundred and eighty nine drug resistant strains of Pseudomonas isolated in hospitals, from antibiotic production sewage and mine soils were studied in experiments on colony hybridization with the use of 32P-labeled plasmid DNA fragments containing various antibiotic resistance genetic determinants. The genes controlling production of aminoglycoside-3'-phosphotransferase, aminoglycoside-3'-adenilyl transferase, type I dihydropteroate synthetase (DHPS I) and DHPS II were detected in the strains of all the ecological niches. More than 90 per cent of the clinical strains hybridized with the samples containing these genes, though in the soil strains the main mechanisms of streptomycin and sulfanilamides resistance were probably controlled by the other nonhomologous genetic determinants studied. The gene controlling production of aminoglycoside 3'-phosphotransferase II was detected in the strains isolated both in the hospitals (49 per cent) and from the sewage (45 per cent), while the gene of aminoglycoside-3'-phosphotransferase I was detected only in the population of the clinical strains (52 per cent). The majority of the tetracycline resistant strains isolated in the hospitals and from the sewage hybridized with the samples containing the classes A and C tetracycline resistance genetic determinants. In the soil strains the genetic determinants of the main tetracycline resistance mechanism(s) were not homologous to those of classes A, B and C. Resistance of the strains to trimetoprine was as a rule determined by the genes of type II dihydrofolate reductase. PMID- 3037996 TI - [Regression analysis in the study of dynamics of 5-nucleotidase activity in mouse peritoneal exudate macrophages]. AB - The time course of the activity of 5-nucleotidase of peritoneal exudate macrophages (PEM) was studied on mice. Regression models of in time changes in the activity of PEM 5-nucleotidase after intraperitoneal administration of 2 immunostimulators: salmosan and blastolysine were constructed by the results of the study. Within the experimentally chosen periods of the study the time changes in the enzyme activity after administration of either salmosan or blastolysine could be adequately approximated by linear regression. The models enabled predicting the enzyme activity levels at various moments within wide ranges after administration of the immunostimulators. Moreover, it is recommended that the regression models of changes in 5-nucleotidase activity after intraperitoneal administration of the immunostimulators be used for quantitative comparison of their efficacy. PMID- 3037997 TI - E-0702, a new cephalosporin, is incorporated into Escherichia coli cells via the tonB-dependent iron transport system. AB - E-0702, a new cephalosporin with a potent antipseudomonal action, was synthesized. In the study of the mode of action of this antibiotic in Escherichia coli, it was found that mutants which acquired resistance to E-0702 were isolated spontaneously and could be shown to be susceptible to its closely related derivatives, E-0702-060 and E-0702-061, and other representative beta-lactam antibiotics. In these mutants, no increased production of beta-lactamase was detectable. No apparent differences between the resistant mutants and the parental strains were observed in the affinity of E-0702 for penicillin-binding proteins. Furthermore, no significant reduction in or loss of both OmpF and OmpC porin proteins in the outer membrane was observed. The mutation was mapped to the tonB gene, which is known to be essential for the iron transport system of bacteria. The bactericidal action of E-0702 was rapidly expressed against iron starved cells in which the iron transport system was induced, whereas the bactericidal action against iron-supplemented cells was ineffective. It is suggested that E-0702 is incorporated into bacterial cells as a chelator of iron via the tonB-dependent iron transport system, after which its strong and rapid bactericidal action is manifested. PMID- 3037998 TI - Pyrrolo[2,3-d]pyrimidine nucleosides as inhibitors of human cytomegalovirus. AB - Seven arabinosyl, 2'-deoxyribosyl, and ribosyl pyrrolo[2,3-d]pyrimidines were evaluated in vitro for activity against human cytomegalovirus and for cytotoxicity in primary and established cell lines of human origin. The parent ribosyl analogs exhibited little antiviral selectivity owing to high cytotoxicity. In contrast, ara-tubercidin, ara-toyocamycin, ara-sangivamycin, and deoxysangivamycin exhibited selectivity between antiviral effect (measured by plaque or titer reduction or both) and cytotoxicity (measured microscopically and by incorporation of radioactive precursors into DNA, RNA, and protein). The selectivity (in vitro therapeutic indexes) for these four compounds ranged from 2 to 40. The two sangivamycin analogs were the most potent and selective. Ara sangivamycin, for example, inhibited virus replication 10(5)-fold at a concentration (10 microM) which produced only partial inhibition of cell growth and labeled precursor incorporation. The four arabinosyl and deoxyribosyl nucleosides appeared to act by inhibition of viral DNA synthesis as quantitated by DNA-DNA dot blot hybridization. These four analogs also were tested for activity against two strains of type 1 herpes simplex virus by a plaque reduction assay. Unexpectedly, all compounds inhibited herpes simplex virus to a lesser extent than human cytomegalovirus. PMID- 3037999 TI - Monocyclic and tricyclic analogs of quinolones: mechanism of action. AB - The mode of action of Ro 13-5478 and Ro 14-9578, monocyclic and tricyclic quinolone analogs, respectively, was examined for Escherichia coli and Staphylococcus aureus. The compounds showed antibacterial activity and effects on cell morphology, replicative DNA biosynthesis, and gyrase-catalyzed DNA supercoiling that were comparable to those shown by nalidixic acid and by oxolinic acid compounds. The results suggest that their site of action is DNA gyrase and that a bicyclic quinolone nucleus is not essential for activity. PMID- 3038000 TI - In vitro activity of zinc salts against human rhinoviruses. AB - The antiviral activity of zinc salts against rhinovirus types 1A and 39 was assayed by yield reduction and inhibition of cytopathic effect in cell culture. The findings indicate that the zinc salts tested have low in vitro therapeutic indices and suggest that the possible beneficial effects of zinc lozenges in reducing cold symptoms may not be related to selective antirhinovirus activity. PMID- 3038001 TI - Antimicrobial activity and spectrum of LY146032, a lipopeptide antibiotic, including susceptibility testing recommendations. AB - LY146032, a new lipopeptide, was found to have a spectrum of gram-positive antimicrobial activity that includes activity against staphylococci (methicillin susceptible and resistant), beta-hemolytic Streptococcus spp., pneumococci, viridans group Streptococcus spp., anaerobic gram-positive cocci, Clostridium spp., and enterococci. The new lipopeptide was generally bactericidal and showed more rapid killing of Listeria spp. (MIC, 1 to 2 micrograms/ml) and staphylococci than either vancomycin or teicoplanin. The 30-micrograms disk was preferred to the 15-micrograms disk on the basis of the preliminary interpretive criteria for susceptibility which indicated zone diameters of greater than or equal to 16 mm for susceptible strains (MIC, less than or equal to 2.0 micrograms/ml) and greater than or equal to 12 mm for resistant strains (MIC, greater than or equal to 8.0 micrograms/ml). These criteria are valid pending the testing of additional gram-positive strains which have LY146032 MICs of greater than or equal to 8 micrograms/ml. PMID- 3038002 TI - Adrenal response to corticotropin during therapy with itraconazole. AB - Itraconazole is a triazole with a mechanism of action similar to that of ketoconazole. Endocrine side effects of ketoconazole, including impaired cortisol synthesis, have been well documented (A. Pont, J. R. Graybill, P. C. Craven, J. N. Galgiani, W. E. Dismukes, R. E. Reitz, and D. A. Stevens, Arch. Intern. Med. 144:2150-2153, 1984). We examined the adrenal response to corticotropin in 10 patients being treated with itraconazole. No impairment of cortisol synthesis could be demonstrated. PMID- 3038003 TI - Sultamicillin in the treatment of superficial skin and soft tissue infections in children. AB - Fifty-two children with superficial skin and soft tissue infections were randomized to receive sultamicillin or cloxacillin for 7 days. Twenty-one children in each group finished the study. A total of 16 of 21 in the sultamicillin group and 13 of 21 in the cloxacillin group were cured. One child in the sultamicillin group and two in the cloxacillin group failed therapy. Four children who received sultamicillin and six who received cloxacillin had recurrences of lesions. Differences were not statistically significant. PMID- 3038004 TI - More than one DNA sequence encodes the 2''-O-adenylyltransferase phenotype. AB - Biochemical and phenotypic assays indicate that three enterobacterial R plasmids isolated in a single hospital encode 2''-O-adenylyltransferase [ANT(2'')], and an ANT(2'')-specific probe was developed from one plasmid. Southern hybridization revealed the three plasmids to be unrelated and, further, showed their ANT(2'') encoding genes to be different. PMID- 3038005 TI - Antimicrobial resistance of Campylobacter jejuni and Campylobacter coli with special reference to plasmid profiles of Japanese clinical isolates. AB - A total of 111 clinical isolates of Campylobacter jejuni and 10 clinical isolates of Campylobacter coli were characterized by their susceptibility to nine antimicrobial agents and by their plasmid profiles on agarose gel electrophoresis. All of the C. jejuni isolates were susceptible to chloramphenicol, ciprofloxacin, erythromycin, kanamycin, and nalidixic acid, but 55% were tetracycline resistant. In the 10 C. coli isolates, a high prevalence of multiple-antibiotic resistance was noted. Plasmids were found in 82% of the tetracycline-resistant and 15% of the tetracycline-susceptible C. jejuni isolates. Tetracycline resistance in six randomly selected C. jejuni isolates, which contained 50- or 135-kilobase (kb) plasmids, was transferred by conjugation to a Campylobacter fetus subsp. fetus recipient with recovery of a 50- or a 45-kb plasmid from transconjugants. From one multiple-antibiotic-resistant C. coli isolate, resistance to tetracycline, kanamycin, and chloramphenicol was transferred concomitantly with a 58-kb plasmid, pNR9589. Nonconjugative 98-kb plasmids, pNR9131 and pNR9581, from C. coli isolates with resistance to tetracycline, kanamycin, and erythromycin were shown by cloning experiments to code for at least kanamycin resistance. Restriction digests revealed that 50-kb plasmids from tetracycline-resistant C. jejuni isolates were identical, although plasmids from multiple-antibiotic-resistant C. coli isolates shared partial DNA homology to each other. Cloning of the kanamycin and chloramphenicol resistance genes of pNR9589 into Escherichia coli showed that the two genes are closely linked or clustered. Double-digestion analysis of the fragments encoding the kanamycin resistance of pNR9131, pNR9581, and pNR9589 showed that these three plasmids contain a common fragment related to kanamycin resistance. PMID- 3038006 TI - Development of natural and synthetic DNA probes for OXA-2 and TEM-1 beta lactamases. AB - Cloning of a 6.3-kilobase BglII DNA fragment from plasmid R46 permitted the isolation of the OXA-2 beta-lactamase gene. Selected DNA fragments internal and adjacent to the OXA-2 beta-lactamase structural gene were used as probes in homology studies with other plasmid-mediated beta-lactamases. Under conditions of high stringency, no cross hybridization could be detected with DNA probes from within the open reading frame of the OXA-2 structural gene. At a lower stringency, one of two DNA fragments used as probes cross hybridized weakly with the OXA-3 bla gene. Other DNA fragments tested and known to contain sequences flanking the OXA-2 determinant cross hybridized with OXA-3 and PSE-4 plasmid DNA. From the known nucleotide sequence of OXA-2 and TEM-1, we synthesized a series of oligonucleotides corresponding to sequences internal to their respective structural genes. A 12-mer oligonucleotide containing the OXA-2-active-site nucleotide sequences cross hybridized only with OXA-3. All other oligonucleotides tested were found to be specific for their respective OXA-2 or TEM-1 gene. Such beta-lactamase gene probes should facilitate studies of beta-lactamase molecular epidemiology and beta-lactamase gene polymorphism. PMID- 3038008 TI - Use of recombinant bovine alpha 1 interferon in reducing respiratory disease induced by bovine herpesvirus type 1. AB - Intranasal or intramuscular treatment of calves with recombinant bovine alpha 1 interferon before challenge with bovine herpesvirus type 1 and Pasteurella haemolytica reduced clinical signs, number of sick days, lung lesions, and weight loss. The effective dose was determined to be relatively broad within the range of 1 to 50 mg per animal. No adverse effects were observed even at high doses of interferon (50 mg per animal). Administration before virus infection was more effective than administration at the same time as virus infection. Although interferon administration had dramatic effects on the survival of animals, it did not have much effect on virus secretion in the upper respiratory tract. Therefore, the mechanism by which interferon reduces the susceptibility of animals to viral-bacterial synergy was postulated to be via its immunomodulatory effects. PMID- 3038007 TI - Molecular epidemiology of macrolides-lincosamides-streptogramin B resistance in Staphylococcus aureus and coagulase-negative staphylococci. AB - Macrolides-lincosamides-streptogramin B (MLS) resistance is commonly found in Staphylococcus aureus and coagulase-negative staphylococci (22 and 45%, respectively, among isolates from three New Jersey hospitals). We have examined representative subsets of 107 MLS-resistant isolates for the molecular nature of the resistance determinant, the erm gene, by dot blot and Southern hybridization analysis. All of 35 S. aureus isolates examined and 39 of 42 coagulase-negative isolates examined were found to harbor the ermA or ermC evolutionary variant. Genes of the ermC class occurred exclusively on a small plasmid similar to or indistinguishable from one (pNE131) previously described in S. epidermidis. Genes of the ermA class occurred exclusively in the chromosome, and restriction patterns indicated that they were part of a transposon, Tn554, characteristic of the classical S. aureus ermA strain. Unlike S. aureus ermA strains examined previously, which harbor Tn554 at a single specific (primary) site, four of our S. aureus isolates had second inserts at different chromosomal sites. The majority of our coagulase-negative isolates had two or more inserts, neither of which occurred at the classical primary site and many of which differed from one another in location (as inferred from restriction patterns). Coagulase-negative staphylococci constitute a large reservoir of the ermA and ermC class of determinants, with clear potential for interspecies spread. PMID- 3038009 TI - Prooxidative activities of 10 phenazine derivatives relative to that of clofazimine. AB - The objective of this study was to investigate the relationship between the antimycobacterial properties of the antileprosy drug clofazimine and its stimulatory effect on the release of reactive oxidants by polymorphonuclear leukocytes by using a variety of phenazine derivatives. The effects of these compounds on myeloperoxidase-mediated iodination, luminol-enhanced chemiluminescence, and the release of superoxide anion by polymorphonuclear leukocytes were investigated. Dissociation of the antimycobacterial and prooxidative effects of clofazimine was possible by manipulation of the chemical group in position 2 of the phenazine molecule. When nitrogen-containing substituents in this position were replaced by oxygen, the mode of the prooxidative action of the compounds was altered. PMID- 3038010 TI - In vitro evaluation of CP-62,993, erythromycin, clindamycin, and tetracycline against Chlamydia trachomatis. AB - The antimicrobial activity of CP-62,993 against four Chlamydia trachomatis isolates was compared with those of erythromycin, clindamycin, and tetracycline. The MIC of CP-62,993 was 0.26 to 1.02 micrograms/ml for 100% inclusion inhibition and 0.031 to 0.063 microgram/ml for 50% inclusion inhibition. With pharmacokinetic and antimicrobial studies demonstrating prolonged half-life and in vitro effectiveness, CP-62,993 may make possible a single, short-course treatment regimen for C. trachomatis infection. PMID- 3038011 TI - Location on RP4 of a tellurite resistance determinant not normally expressed in IncP alpha plasmids. AB - The tellurite resistance (Ter) determinant of RP4 is not normally expressed unless variants are selected on medium containing tellurite. The determinant was mapped in the variant plasmid RP4Ter by Tn7 insertion mutagenesis. Based on a 56.4-kilobase (kb) replicon, it covered the region from 56 kb, across the EcoRI site at 0 kb, to 1.5 kb. PMID- 3038012 TI - Susceptibility of anaerobic bacteria to FCE 22101. AB - The activity of FCE 22101 against 675 strains of anaerobic bacteria was determined by an agar dilution method. Its activity was compared with those of benzylpenicillin, piperacillin, cefoxitin, imipenem, clindamycin, metronidazole, chloramphenicol, vancomycin, fusidic acid, and bacitracin. FCE 22101 and imipenem (MIC, less than or equal to 0.5 mg/liter) were the most active agents against the anaerobic strains tested, except against Clostridium difficile strains. On the basis of these results, FCE 22101 appears to be a promising antimicrobial agent for treatment of anaerobic infections, and further clinical investigations are indicated. PMID- 3038013 TI - Synergistic action of quercetin and murine alpha/beta interferon in the treatment of Mengo virus infection in mice. AB - ABD2F1 mice were infected intraperitoneally (i.p.) or intranasally (i.n.) with Mengo or Sindbis virus and treated with either crude murine alpha/beta interferon (MuIFN-alpha/beta) or quercetin, or both. MuIFN-alpha/beta given i.p. or intramuscularly (i.m.) 1-3 h before the infection had a dose-dependent protective effect regardless of the route of administration. When given after the infection, IFN did not show any effect. Oral quercetin, capable of protecting cardio, i.e. Mengo virus-infected mice, failed to show antiviral efficacy in Sindbis virus infected animals. Of various combinations of quercetin and MuIFN-alpha/beta, a certain well defined regimen resulted in a significant enhancement of protection in Mengo, but not Sindbis, virus-infected mice. A marginally effective treatment regimen of quercetin (20 mg/kg, given 12 h before Mengo virus infection, and 10 mg/kg given both 1 h before and 12 h after infection) potentiated the activity of a single dose of MuIFN-alpha/beta (5000 IU 3 h prior to infection), giving 85 100% survivors compared to 50% for MuIFN-alpha/beta when applied alone (p less than 0.001). PMID- 3038014 TI - Generation of Tn5 insertions in streptococcal conjugative transposon Tn916. AB - A method has been developed for the introduction of Tn5 into Escherichia coli plasmid chimeras containing Streptococcus faecalis DNA. Tn5 could be introduced via a lambda::Tn5 delivery vehicle. The system proved to be particularly efficient and facilitated insertions at numerous sites on DNA containing the 16 kilobase conjugative transposon Tn916. It was possible to introduce some of the resulting Tn916::Tn5 derivatives back into S. faecalis by using a recently developed protoplast transformation procedure. A presumed zygotic induction resulted in insertion of the Tn916 derivatives at multiple sites in the S. faecalis chromosome. PMID- 3038015 TI - Restriction enzyme analysis of lactose and bacteriocin plasmids from Streptococcus lactis subsp. diacetylactis WM4 and cloning of BclI fragments coding for bacteriocin production. AB - The 131.1-kilobase (kb) bacteriocin production (Bac) plasmid pNP2 and the 63.6-kb lactose metabolism (Lac) plasmid pCS26, from Streptococcus lactis subsp. diacetylactis WM4, as well as pWN8, a 116.7-kb recombinant plasmid from a Lac+ transconjugant, were analyzed with restriction enzymes to determine the origin of pWN8. Plasmid pWN8 conferred a Lac+ Bac- phenotype, contained DNA derived from pCS26 and pNP2, and, like pNP2, exhibited self-transmissibility (Tra+). In cloning attempts, Bac+ transformant S. lactis KSH1 was isolated. The recombinant plasmid, pKSH1, contained three BclI fragments from pNP2. Bac- transformants which individually contained each of the three fragments were also identified. Comparison of restriction maps of pKSH1 and pNP2 revealed an 18.4-kb region common to both plasmids, involving two of the three BclI fragments. S. lactis KSH1 also exhibited greater inhibitory activity against the indicator strain S. diacetylactis 18-16 than did a strain containing the 131.1-kb Bac plasmid. PMID- 3038016 TI - An A-ELISA to detect hepatitis A virus in estuarine samples. AB - An amplified enzyme-linked immunosorbent assay (A-ELISA) for detecting and quantifying hepatitis A virus in estuarine water samples is described. The test was five times more sensitive than a standard ELISA and at least two times more sensitive than radioimmunoassay. Test sensitivity was unaffected by the procedures used to concentrate the virus in estuarine samples or by the presence of humic and tannic acids in test samples. Nonspecific reactions were not encountered with a number of enteroviruses or with a rotavirus. A high sensitivity and specificity combined with speed, low cost, and freedom from radiolabels made the A-ELISA useful for detecting hepatitis A virus in environmental samples. The virus was detected in 3 of 20 estuarine water samples examined by A-ELISA. PMID- 3038017 TI - Rapid plasmid analysis for identification of Edwardsiella ictaluri from infected channel catfish (Ictalurus punctatus). AB - Eighteen different strains of Edwardsiella ictaluri isolated from infected channel catfish (Ictalurus punctatus) were screened to determine whether plasmid DNA was present. Two plasmids of 5,700 and 4,900 base pairs were identified. Restriction enzyme analysis showed that each of the strains harbored these same two plasmids. Restriction maps of the separated plasmids indicated that these plasmids were not closely related to each other. A rapid screening technique was developed that would allow the presence of these plasmids from either broth cultures or single colonies of E. ictaluri to be determined within 2 to 3 h by agarose gel electrophoresis. These results suggest that plasmid fingerprinting of E. ictaluri should become a useful tool in the presumptive identification of this bacterium from infected channel catfish. PMID- 3038018 TI - Ionization of clusters. III. Evaluation of interaction parameters from binding data. AB - Interactions between ionizable groups on the same molecule modulate the binding of protons to an extent where the binding constants may be shifted by orders of magnitude. The first two papers of this series discussed the family of carboxylic acids, pairwise isotropic interactions, and evaluation of single site binding data. This paper presents an extended group of hypothetical binding isotherms. Procedures are illustrated for deriving interaction parameters from binding data. The interaction parameters for about 25 representative compounds with two and three interacting ionizable groups are evaluated and tabulated. PMID- 3038019 TI - Comparison of thermal properties of bovine spectrin and fodrin. AB - Thermal properties of bovine brain fodrin have been studied by circular dichroism and electron spin resonance and compared to those of bovine erythrocyte spectrin. Protein unfolding was induced either by urea or by a combination of heat and urea. The denaturation profiles of the two proteins, as measured by the changes in ellipticity at 222 nm as a function of temperature, were very similar but fodrin denaturation occurred at both higher temperatures and higher urea concentrations. In the absence of urea the thermal transition of spectrin was centered at 51 degrees C and that of fodrin at 54.5 degrees C. Proteins were also labeled with a maleimide analog spin probe. Spin-labeled fodrin showed a thermal transition similar to that of spectrin but centered at 46 degrees C instead of 42 degrees C. These findings indicated a close structural similarity of the two proteins but a slightly higher conformational stability of fodrin to both heat and urea. PMID- 3038020 TI - Changes in pulmonary calpain activity following treatment of mice with butylated hydroxytoluene. AB - The antioxidant, butylated hydroxytoluene (BHT), causes lung toxicity in mice followed by regenerative repair, and can also modulate the development of carcinogen-induced lung adenomas. We are investigating changes in pulmonary biochemistry following BHT treatment in order to understand the mechanisms of BHT induced pulmonary regenerative repair. BHT administration lowered cytosolic Ca2+ activated neutral protease (calpain) activity, increased the activity of the endogenous calpain inhibitor, calpastatin, increased the extent of photoincorporation of 8-N3-[32P]cAMP into a Mr 37,000 proteolytic product derived from cAMP-dependent protein kinase regulatory (R) subunits, and increased membrane-associated protease activity. All of these changes were dependent on the BHT dosage; the altered proteolytic activities occurred at a dose lower than that which caused observable lung toxicity as assessed by the lung weight/body weight ratio. Decreased cytosolic calpain activity was detectable within 1 day after BHT administration, was lowest at 4-7 days, and had not returned to control levels by Day 21, a time when normal lung morphology had been regained. The decrease in calpain activity cannot fully be accounted for by increased calpastatin activity; upon separation of these proteins by DEAE chromatography, the amount of calpain activity from BHT-treated mice remained lower than the corresponding peak from control mice. Increased photolabeling of the Mr 37,000 protein began at 1 day and continued to increase up to 4 days after BHT. All of the cytosolic changes preceded the increased particulate proteolytic activity by 1-2 days. R-subunits which have dissociated from their catalytic subunits are more susceptible to degradation by calpain, but BHT treatment did not enhance subunit dissociation as determined by the elution profile of 8-N3-[32P]cAMP-labeled R-subunits following DEAE chromatography. A large percentage of the particulate protease activity was inhibited by calpastatin, leupeptin, and E-64, all of which are known to inhibit calpain activity; this suggested that calpain accounted for most of this activity. Changes in the activities of proteases which catalyze limited proteolysis reactions may play an important role in the repair of acute lung injury. PMID- 3038021 TI - Reduction and destruction rates of nitroxide spin probes. AB - A series of nitroxides was tested for rates of one-electron reduction in a chemical, a photochemical, and two biological systems by ESR assays. In all cases, piperidine and hydropyridine nitroxides were reduced consistently more rapidly than pyrroline and pyrrolidine nitroxides. Substituents on the nitroxides also affected reduction rates, although not as greatly as ring structure. One of the reduction systems, consisting of the photosensitizer FMN and the photoreductant EDTA, was used to study both anaerobic reduction and O2-dependent reoxidation of some of the nitroxides. Reduced piperidine and hydropyridine nitroxides were also oxidized more rapidly than the reduced pyrroline and pyrrolidine nitroxides. Reoxidation subsequent to reduction was partially inhibited by superoxide dismutase, indicating that superoxide radicals are involved in the process. Even after prolonged reoxidation, not all of the probe molecules were returned to their oxidized form, implying an irreversible "destruction" of the spin probe concomitant with its chemical reduction. Probe destruction was studied more specifically with a photochemical system for generating methyl radicals, which showed that these carbon-centered radicals destroyed different nitroxides at rates which were much less influenced by the nitroxide structures than one-electron reduction was. PMID- 3038022 TI - Cilastatin-sensitive dehydropeptidase I enzymes from three sources all catalyze carbapenem hydrolysis and conversion of leukotriene D4 to leukotriene E4. AB - The dipeptidase, dehydropeptidase I (EC 3.4.13.11), was purified to homogeneity from rat lung, rat kidney, and hog kidney. Analysis of physical parameters (subunit molecular weights, Km values for glycyldehydrophenylalanine, Ki values for dehydropeptidase I inhibitors, and immunoreactivity) showed the rat dipeptidases to be similar to each other but different from the hog dipeptidase. However, all three enzymes hydrolyzed imipenem and converted leukotriene D4 to leukotriene E4, and these activities were inhibited by cilastatin. PMID- 3038023 TI - Molecular properties of pyrophosphate:fructose-6-phosphate phosphotransferase from potato tuber. AB - Pyrophosphate:fructose-6-phosphate phosphotransferase (PFP) from potato tubers has been purified to homogeneity. The enzyme contains two polypeptides with apparent relative molecular mass (Mr) values of 65,000 and 60,000. These polypeptides give different peptide fragments after limited proteolytic digestion. Antibodies raised against each polypeptide separately are specific for that polypeptide, but both antisera are capable of immunoprecipitating native PFP activity. These antibodies also recognize similar pairs of polypeptides in a range of other plant tissues that contain PFP activity. Based on gel filtration, the Mr value of potato tuber PFP is 265,000. This suggests that the enzyme is a heterotetramer composed of two polypeptides with Mr values of 65,000 and 60,000. In the presence of pyrophosphate, potato PFP dissociates into a 130,000 dimer. PMID- 3038024 TI - Overproduction, purification, and characterization of adenylosuccinate synthetase from Escherichia coli. AB - Adenylosuccinate synthetase, encoded by the purA gene of Escherichia coli, catalyzes the first committed step toward AMP in the de novo purine biosynthetic pathway and plays an important role in the interconversion of purines. A 3.2-kb DNA fragment, which carries the purA gene, was cloned into the temperature inducible, high-copy-number plasmid vector, pMOB45. Upon temperature induction, cells containing this plasmid produce adenylosuccinate synthetase at approximately 40 times the wild-type level. A scheme is presented for the purification of the overproduced adenylosuccinate synthetase to homogeneity in amounts sufficient for studies of its structure and mechanism. The wild-type and the overproduced adenylosuccinate synthetase enzyme preparations were judged to be identical by the following criteria. The amino acid sequence at the N-terminus of the overproduced enzyme proved identical to the corresponding sequence of the wild-type enzyme. Michaelis constants for both the wild-type and overproduced enzyme preparations were the same. And (iii) both proteins shared similar chromatographic behavior and the same mobility during sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. Results from size-exclusion chromatography and SDS-polyacrylamide gel electrophoresis suggest that adenylosuccinate synthetase exists as a dimer of identical, 48,000-Da, subunits. PMID- 3038025 TI - Regulation of mRNA levels for five urea cycle enzymes in rat liver by diet, cyclic AMP, and glucocorticoids. AB - Adaptive changes in levels of urea cycle enzymes are largely coordinate in both direction and magnitude. In order to determine the extent to which these adaptive responses reflect coordinate regulatory events at the pretranslational level, measurements of hybridizable mRNA levels for all five urea cycle enzymes were carried out for rats subjected to various dietary regimens and hormone treatments. Changes in relative abundance of the mRNAs in rats with varying dietary protein intakes are comparable to reported changes in enzyme activities, indicating that the major response to diet occurs at the pretranslational level for all five enzymes and that this response is largely coordinate. In contrast to the dietary changes, variable responses of mRNA levels were observed following intraperitoneal injections of dibutyryl cAMP and dexamethasone. mRNAs for only three urea cycle enzymes increased in response to dexamethasone. Levels of all five mRNAs increased severalfold in response to dibutyryl cAMP at both 1 and 5 h after injection, except for ornithine transcarbamylase mRNA which showed a response at 1 h but no response at 5 h. Combined effects of dexamethasone and dibutyryl cAMP were additive for only two urea cycle enzyme mRNAs, suggesting independent regulatory pathways for these two hormones. Transcription run-on assays revealed that transcription of at least two of the urea cycle enzyme genes -carbamylphosphate synthetase I and argininosuccinate synthetase--is stimulated approximately four- to fivefold by dibutyryl cAMP within 30 min. The varied hormonal responses indicate that regulatory mechanisms for modulating enzyme concentration are not identical for each of the enzymes in the pathway. PMID- 3038026 TI - 23Na and 31P NMR studies of the effects of salt stress on the freshwater cyanobacterium Synechococcus 6311. AB - We have used 23Na and 31P nuclear magnetic resonance (NMR) spectroscopy to elucidate some of the bioenergetic changes that occur in the freshwater cyanobacterium Synechococcus 6311 after a transition from growth medium (Na concentration 0.01 M) to medium containing 0.5 M NaCl. 23Na NMR analysis showed Na rapidly penetrates the cells under dark aerobic conditions; cells grown for several days in high salt medium, however, reestablish a low internal sodium content, comparable to control cells. For 31P NMR analysis, a system was devised to aerate and illuminate cell suspensions during spectral acquisition. The NMR spectra showed that when cells are presented with 0.5 M NaCl (final concentration), nucleotide triphosphate peaks decrease, the inorganic phosphate peak increases, and the cytoplasmic pH transiently increases from 7.4 to 7.9. Pyrophosphate added to cell suspensions is hydrolyzed to inorganic phosphate apparently by an extracellular phosphatase, allowing external and internal pools of inorganic phosphate to be distinguished. Nucleotide triphosphate levels fall almost as much when cells are incubated in darkness as under anoxia, indicating that both respiration and photosynthesis contribute to the maintenance of intracellular ATP levels. Cells grown in high salt medium for several generations exhibited a pattern of 31P metabolites similar to control cells, except that they produced more (and more intense) peaks in the monoester phosphate region, presumably signals from sugar phosphates. PMID- 3038027 TI - Spinach leaf ribulose-5-phosphate kinase: examination of sulfhydryls by chemical modification and spin-labeling. AB - Methodology has been developed for complete or selective modification of the cysteinyl sulfhydryls of ribulose-5-phosphate (Ru5P) kinase. Using native enzyme, iodoacetate modifies four sulfhydryls with varying levels of completeness. The most reactive sulfhydryl in the native enzyme can be selectively titrated with iodoacetate; complete loss of activity occurs. Composition and N-terminal analyses of the peptide bearing this essential sulfhydryl indicate that the alkylated residue (Cys-16) is identical to the site modified by other modification reagents (M. A. Porter and F. C. Hartman (1986) Biochemistry 25, 7314-7318). In the presence of ATP, a nonessential sulfhydryl of the native enzyme is carboxymethylated. The peptide bearing this modified cysteine has been isolated and its composition and N-terminal sequence determined. Enzyme that is carboxymethylated in the presence of ATP retains activity and can be oxidatively inactivated in a reversible fashion. This suggests that the cysteine targeted by iodoacetate in the presence of ATP is not a residue that participates in regulation of enzyme activity. Using a spin-labeled analog of iodoacetate, both essential and nonessential cysteines have been selectively modified. ESR measurements suggest that the environment of these cysteines is not highly constrained. Modest effects on spin-label mobility are observed upon occupancy of Ru5P or ATP sites on the modified enzyme. These effects are dependent on the presence of divalent cations, suggesting that a binary enzyme-cation complex must form prior to productive enzyme-substrate interactions. PMID- 3038028 TI - Oxygen radicals are generated by dye-mediated intracellular photooxidations: a role for superoxide in photodynamic effects. AB - Representative thiazines, xanthenes, acridines, and phenazines photosensitized the oxidation of reduced pyridine nucleotides and reduced glutathione when illuminated with low intensity visible light. Photooxidation resulted in oxygen consumption and in superoxide generation, assayed as the superoxide dismutase (SOD)-inhibitable reduction of ferricytochrome c. The major pathway of electron transfer involved dye reduction rather than singlet oxygen-mediated oxidation of the substrate, as demonstrated by the relative insensitivity of the oxidation to inhibition by sodium azide and by the observable bleaching of the dye. Hydrogen peroxide was a stable end product of photooxidation. Photosensitive dyes were photoreduced intracellularly. These dyes were transported across the membranes of Escherichia coli B and stimulated a light- and concentration-dependent increase in the cyanide-insensitive respiration. Dyes reduced intracellularly subsequently diffused out of the cell where they reduced extracellular cytochrome c. The photosensitive dyes examined in this study exhibited a light-dependent bacteriostatic effect on E. coli B grown in nutrient broth, manifested as an increased lag prior to growth. Restoration of growth coincided with increased levels of SOD, and the intracellular level of SOD correlated with the level of illumination, the dye concentration, and the reactivity of the dye to NADH in vitro. The thiazine dye, toluidine blue o, imposed a light- and oxygen-dependent lethality on E. coli B grown in glucose minimal medium. Toxicity was relieved by hydroxyl radical scavengers, and their ability to protect the cells was proportional to their reactivity with the hydroxyl radical. The results indicate that oxygen radicals and related species mediate photodynamic effects in E. coli B. PMID- 3038029 TI - Biosynthesis of the sesquiterpene patchoulol from farnesyl pyrophosphate in leaf extracts of Pogostemon cablin (patchouli): mechanistic considerations. AB - Several mechanistic alternatives have been proposed for the enzyme-catalyzed, electrophilic cyclization of farnesyl pyrophosphate to the tricyclic sesquiterpene alcohol patchoulol, which is the characteristic component of the essential oil of Pogostemon cablin (patchouli). These alternatives include schemes involving deprotonation-reprotonation steps and the intermediacy of the monocyclic and bicyclic olefins germacrene and bulnesene, respectively, and involving a 1,3-hydride shift with only tertiary cationic intermediates and without any deprotonation-reprotonation steps. Analytical studies, based on analyses of P. cablin leaf oil at different stages of plant development, and in vivo time-course investigations, using 14CO2 and [14C]sucrose, gave no indication that germacrene and bulnesene were intermediates in patchoulol biosynthesis. A soluble enzyme system from P. cablin leaves was prepared, which was capable of converting farnesyl pyrophosphate to patchoulol, and isotopic dilution experiments with both labeled and unlabeled olefins were carried out with this system to confirm that sesquiterpene olefins did not participate as fre intermediates in the transformation of the acyclic precursor to patchoulol. Patchoulol derived biosynthetically from [12,13-14C;1-3H]farnesyl pyrophosphate was chemically degraded to establish the overall construction pattern of the product. Similar studies with [12,13-14C;6-3H]farnesyl pyrophosphate as a precursor eliminated deprotonation steps to form bound olefinic intermediates in the biosynthesis of patchoulol, while providing supporting evidence for the hydride shift mechanism. PMID- 3038030 TI - Frontal gel chromatography of interacting systems: theoretical and experimental evaluation of the shapes of elution profiles for systems of the type A + B in equilibrium C. AB - A theoretical and experimental study has been made of the advancing elution profile in frontal gel chromatography of interacting systems for which the elution volume of the complex is smaller than that of the larger reactant. First, Gilbert-Jenkins theory is used to delineate the form of the elution profile from the magnitudes of the elution volumes and concentrations of reacting species. This procedure resulted in the detection of a misinterpretation of certain patterns obtained in a gel chromatographic study of the interaction between myoglobin and ovalbumin. Second, a numerical computational procedure, which incorporates both axial dispersion and concentration-dependence of species elution volumes, is used to establish the influence of these two factors on boundary shapes for such systems. Third, frontal gel chromatography on Sephadex G 75 is used to compare experimental behavior with theoretical profiles predicted for the electrostatic interaction between cytochrome c and soybean trypsin inhibitor (pH 6.8, I 0.01). Results of these experiments serve as a guide for future conduct of experiments aimed at characterization of biologically important, reversible complex formation between proteins and/or other macromolecules. PMID- 3038031 TI - [Liver phosphotyrosine protein phosphatase and tyrosine protein kinase in chemical hepatocarcinogenesis]. AB - We found and characterized three forms of phosphotyrosine protein phosphatase, subsequently designated PTPP-1, -2 and -3, respectively, in rat liver. Chemical hepatocarcinogenesis according to Solt and Farber was accompanied by a slight increase in liver phosphotyrosine protein phosphatase activity and a remarkable increase in liver tyrosine protein kinase activity. A maximum 8-fold increase in tyrosine kinase activity was observed in hepatomas induced with 3'-methyl-4 dimethylaminoazobenzene (MeDAB). Tyrosine protein kinase that increased with the progress of chemical hepatocarcinogenesis was solubilized from the particulate fraction of MeDAB-induced hepatoma. The enzyme was shown to require Mg2+ for its activity, to immunoprecipitate with anti-pp-60src-IgG and to phosphorylate the IgG. Rat liver also contains another tyrosine protein kinase which requires Mn2+ and does not immunoprecipitate with anti-pp60src; the level of this enzyme appears to diminish with the progress of chemical hepatocarcinogenesis. PMID- 3038032 TI - [Anti-tumor effect and mechanism of action of the tumor necrosis factor (TNF)]. AB - TNF is cytokine derived from macrophages and shows much promise for use in cancer therapy because of its marked antitumor effects and its high specificity to tumors. The clinical application (Phase I-II) of TNF has been started because human recombinant TNF (rH-TNF) can be produced on a large scale. In spite of notable antitumor effects, little is known concerning the mechanism of its cytotoxic action. In this article, the antitumor effects of rH-TNF against human and murine tumors, the mechanism of its action and the synergistic effects of rH TNF in combination with IFN-gamma, various anticancer drugs or with hyperthermia are reviewed. PMID- 3038033 TI - [Hepatitis B virus integration and oncogene activation]. AB - In hepatitis B virus (HBV) related hepatoma samples there is a high frequency of HBV-DNA integration into the chromosome, but the frequency of integration in the early-stage of carcinogenesis is expected to be even higher. Structural an analysis of integrated HBV-DNA and chromosomal flanking DNAs disclosed that the marked rearrangement of chromosomal DNA is not directly linked to carcinogenesis. HBV-DNA integration and chromosomal DNA depletion occur even in chronic hepatitis tissues. Integration is, therefore, thought to trigger a series of reactions which lead to carcinogenesis. However, the oncogene is not detected within the HBV genome, and the HBV integration site in the chromosome is variable. As one approach to clarifying the cause and effect relationship between HBV-DNA integration and liver carcinogenesis DNA transfection experiments were conducted using mouse NIH3T3 cells to detect hepatoma-related oncogenes. As a result, the well-known N-ras gene and an unknown oncogene were independently isolated from different hepatoma tissues. PMID- 3038034 TI - [A novel oncogene from a human hepatocellular carcinoma]. AB - Primary human hepatocellular carcinomas (HCC) were surveyed for oncogenes by transformation assays using NIH3T3 cells and the calcium phosphate coprecipitation method. High-molecular-weight DNA obtained from the HCC tissues was employed for this purpose. One new transforming DNA, named lca (for liver cancer) was obtained, and its properties were studied in detail. This transforming DNA had a linkage to the Alu sequence and was cloned in lambda phage. Restriction enzyme analyses showed that the minimum size of the lca DNA is about 10.5 kilobase pairs and that its nucleotide sequence is different from that of any known human gene or retroviral oncogene. Southern blot analyses of DNAs from flow-sorted human chromosomes and human-mouse somatic cell hybrids indicated that the lca DNA is located on human chromosome 2. It showed identical restriction enzyme cleavage profiles with its counterpart DNA obtained from normal tissue, indicating that there is no extensive DNA rearrangement associated with the activation process of the lca DNA. The normal counterpart DNA shows no transforming activity. An independently obtained transforming DNA from another HCC exhibited identical restriction enzyme cleavage profiles. Thus, lca DNA is likely to represent a commonly encountered transforming DNA in HCC. PMID- 3038035 TI - [Attempts to improve hybridoma technology for the production of human monoclonal antibodies]. AB - The technology for producing hybridomas secreting human monoclonal antibodies has not been well established, contrary to the situation for mouse monoclonal antibodies. Therefore, we attempted to find better fusion partners using two EBV transformed human B cell lines, C5TK1 and TAPC-30 (anti-SRBC IgM and anti-HBs IgG secretors, respectively; provided by Prof. Y. Ono, Nihon Univ. School of Med.). Fifteen partner cell lines available were fused with either of these lines by conventional PEG technique, and their fusion frequencies, cloning efficiencies, and levels and stabilities of specific antibody secretion were examined. Among the fusion partners tested, KR-12, SHM-D33 and 3HL3-6J lines appeared to be the best. Hybridomas with the KR-12 line were easily cloned and produced stable specific antibodies, although the level of Ig secretion was almost the same as the antibody-producing parental B-cell lines. SHM-D33 cells efficiently formed hybrids but the stability of their Ig production was low and cloning was laborious. Although the fusion frequency of 3HL3-6J cells was not so high, some hybrids were good producers of antibodies, and the levels of specific antibody reached levels greater than 10 micrograms/ml. Their cloning was relatively easy. Therefore, the 3HL3-6J line seemed to be the best fusion partner for EBV transformed B cells. With regard to the low fusion frequency of human cells, we attempted to improve the efficiency using the electrofusion technique. Three to 10 times more hybridomas were obtained by this technique than by the conventional PEG method. Further investigations are under way. PMID- 3038036 TI - Controlled study of exclusion of dietary vasoactive amines in migraine. AB - To assess the effects of dietary vasoactive amines in the aetiology of childhood migraine, 39 children were randomly allocated to either a high fibre diet low in these substances or a high fibre diet alone. Both groups of children showed a significant decrease in the number of headaches and there was no significant difference between the two groups. Dietary vasoactive amines have not been shown in this study to influence childhood migraine. The improvement seen in both groups emphasises the need for a control diet in studies designed to show that dietary manipulation improves disease. PMID- 3038037 TI - [Lesions of the nervous system in AIDS]. PMID- 3038038 TI - [Fatal neurotoxoplasmosis and cytomegalovirus encephalitis in AIDS. Apropos of 2 cases]. PMID- 3038039 TI - Human liver regeneration after major hepatic resection. A study of normal liver and livers with chronic hepatitis and cirrhosis. AB - The regenerative process was evaluated in terms of liver size, function, and histology in 28 adults who had major hepatic resection: hepatocellular carcinoma (HCC) in 21, secondary liver cancer from colorectum in four, carcinoma of the gallbladder in one, Klatskin tumor in one, and Caroli's disease in one. There were 22 men and six women. Ages ranged from 17 to 74 years with a mean age of 56.7. All patients with HCC had underlying liver disease: liver cirrhosis in 14 and chronic hepatitis in seven. Extended right lobectomy was done on 10 patients, right lobectomy on 16 patients, and left lobectomy on two patients. The residual liver size was serially estimated with computed tomography (CT) in 15 patients: six with normal liver, five with chronic hepatitis, and four with cirrhosis. A complete restoration of the residual liver size was found within 3 months in 3 and 6 months, respectively, in two patients with normal livers. The liver was enlarged in all patients with the parenchymal diseases but obviously more slowly compared with normal liver. Liver functions were restored normally within 2-3 weeks in patients with normal livers, but hyperbilirubinemia persisted longer in those with chronic hepatitis and cirrhosis. A continuous rise of bilirubin was an ominous sign of liver failure and subsequent death, which occurred in five patients with cirrhosis. Serum alpha-fetoprotein did not rise in accordance with the regeneration. Histologically, evidence of active regeneration with increased mitotic activity was found at 10 and 35 days in those patients with normal livers. Mitosis was not seen in a specimen taken at 7 days. Enlarged cuboidal hepatocytes and cells with basophilic cytoplasm or two nuclei were observed more or less in all specimens. The livers with cirrhosis or hepatitis also showed histologic evidence of regeneration during the first 2 months but substantially less compared with normal liver, which was well supported by the volumetric study of the liver remnants with CT. PMID- 3038040 TI - Regional therapy in the management of intrahepatic recurrence after surgery for hepatoma. AB - The significance of regional therapy against the intrahepatic recurrence for hepatocellular carcinoma (HCC) was evaluated. During the past 6 years, 101 patients who had radical operations for HCC (liver cirrhosis, 75%; chronic hepatitis, 22%) were followed. Forty-seven patients (47%) had recurrences; the liver was the site of first recurrence in 43 patients and distant site recurrence in four patients. In the patients where the liver was the recurrent site, 33 patients (77%) received regional therapies for an aggregate total of 60 times. Transcatheter arterial chemoembolization was chosen as the first preferred therapy against recurrence in the liver in 30 of 33 patients (91%). Postrecurrence survival of the patients treated with regional therapy was significantly better than disease-free survival (p less than 0.001). Disease-free survival after surgery, postrecurrence survival, and postsurgery survival were compared in the patients treated with regional therapy and untreated patients. Fifty per cent survival after recurrence of the treated patients was 27 months, and that of the untreated patients was 3 months. Postrecurrence survival (p less than 0.001) and postsurgery survival (p less than 0.01) of the treated patients were significantly better than those of the untreated patients. To obtain successful long-term survival after surgery for the cirrhotic patients with HCC, it is necessary to repeat active regional therapies against the recurrent liver. PMID- 3038041 TI - Delivery of intraoperative radiation therapy after pneumonectomy: experimental observations and early clinical results. AB - Intraoperative radiation therapy (IORT) is capable of delivering high doses of radiation to mediastinal structures while sparing lung parenchyma, heart, and other locoregional tissues. A canine model of pulmonary resection and IORT was investigated by performing a pneumonectomy in 15 adult foxhounds followed by 0 cGy, 2,000 cGy, 3,000 cGy, 4,000 cGy. No clinical complications developed in 4 animals in the 2,000-cGy group. However, 2 of the 8 animals given a high dose died of esophageal hemorrhage or carinal necrosis. Esophagitis occurred in 10 of 12 animals, and none of the animals experienced bronchial stump dehiscence. In a limited Phase I protocol, 4 patients with non-small cell lung cancer were treated with resection and 2,500 cGy of IORT to two separate ports encompassing the superior and inferior mediastinum. Two patients experienced life-threatening bronchopleural fistulas, and 2 patients died as a consequence of esophageal problems. One patients had recurrence with brain metastases, and the 1 long-term survivor is free from disease. As opposed to the animal model of thoracic IORT, the clinical study demonstrated major toxicity with respiratory and esophageal morbidity. The therapeutic usefulness of thoracic IORT in the management of lung cancer must be questioned in view of this small but consistent series of patients. Further carefully designed clinical studies using lower doses of IORT are needed. PMID- 3038042 TI - [Various considerations on the current use of nuclear cardiology studies]. PMID- 3038043 TI - [Characterization of an endogenous factor with digitalis-like activity in the heart of the guinea pig (I): Inhibition of Na,K-ATPase)]. AB - The purpose of this work is to study the presence of an endogenous inhibitor of the sodium pump in mammals. A water extract was prepared from guinea pig heart and partially purified by liquid chromatography. The extract inhibited the activity of Na,K-ATPase but had no effect on Mg-ATPase nor on Ca-ATPase. The inhibition was similar to that produced by cardiac glycosides since it was dose dependent, antagonised by potassium and the Na,K-ATPase prepared from brain was more sensitive than the one from heart. These results support the hypothesis on an endogenous factor exhibiting digitalis-like activity that could be a physiological regulator of the sodium pump. PMID- 3038044 TI - Basal ganglia influences on the cerebellum of the cat. AB - The changes in firing rate of intracerebellar nuclear neurons following electrical stimulation of the contralateral basal ganglia were investigated in adult cats, in which antidromic activation of cortico-pontine and/or cortico olivar fibers arising in the area 6 had been excluded by chronic ablation of the motor cortex. Activation of putamen and caudate nucleus induced discharge changes in a low percentage (below 12.5%) of both medial and lateral cerebellar nuclei neurons, while stimulation of globus pallidus and especially of entopeduncular nucleus modified the spontaneous discharge of a greater percent of cells (up to 29%), mainly in the most lateral cerebellar portions. The basal ganglia-induced effects were abolished upon section of the brachium pontis but not of the restiform body. Latency values of the responses, which were predominantly excitatory in nature, suggest the involvement of structures interposed between basal ganglia and precerebellar systems. We postulated that impulses issued by the basal ganglia could reach the cerebellum through a pathway that involves the pedunculopontine nucleus and the nucleus reticularis tegmenti pontis. PMID- 3038045 TI - Axonal projections of the Retzius cells to the dorsal segmental root in the leech Hirudo medicinalis. AB - The presence of RC's axon running in the dorsal root of the leech Hirudo medicinalis was studied by means of electrophysiological and neuroanatomical techniques. Electrical stimulation of the dorsal root gave invariably the antidromic invasion of the Retzius cell. This effect disappears after crushing the dorsal root between the stimulating electrode and the ganglion, and hence is not due to stimulus spread to the nearby posterior root. Anatomical evidence for an axon collateral running in the dorsal root was provided by catecholamine histofluorescence of the axonal branches of the RCs and by Lucifer Yellow intracellular injection into the RCs. PMID- 3038046 TI - Evidence for a nigro-pulvinar-lateralis posterior complex projection in the cat using horseradish peroxidase neuronal retrograde technique. AB - The possible existence of a direct projection from the substantia nigra to the pulvinar-lateral posterior complex (Pul-LP) was investigated in the cat by using the horseradish peroxidase technique. In particular horseradish peroxidase was injected in the Pul-LP of 8 animals, either unilaterally or bilaterally. Tissue sections obtained from the cat's brain 24-48 hrs. after injection were prepared according to Mesulam's method as slightly modified by the authors. Retrogradelly labelled neurons were observed in substantia nigra pars lateralis and reliculata ipsilaterally to the injected pulvinar-lateral posterior complex. A small number of labelled cells were also found in the contralateral substantia nigra. These findings demonstrate the existence of a close connection between two system which are involved in turning behavior: the nigrostriatal and the pulvinar-lateral posterior complex-superior colliculus. PMID- 3038047 TI - [The role of hepatic surgery in hepatoma and ensuing problems]. PMID- 3038048 TI - [The activity of adenosine desaminase, 5'-nucleotidase, and magnesium-, sodium, potassium- and calcium-ATPase in supernatants and homogenates of different tissues and that of adenosine desaminase in the serum of cattle]. PMID- 3038049 TI - [Protection of sheep against infection with bovine leukemia virus by vaccination with tumor cells or tumor cell preparations from lymph nodes of leukemic cattle]. PMID- 3038050 TI - [Pathology and electron microscopic demonstration of viruses in tissues in Derzsy's disease (Parvovirus infection) of goslings]. PMID- 3038051 TI - Characterisation of lymphoproliferative disease virus of turkeys. Structural polypeptides of the C-type particles. AB - Lymphoproliferative disease virus of turkeys (LPDV), a C-type retrovirus, was shown to contain 3 major [32 kilodaltons (kd, p 32), 26 kd, 22/21 kd] and 2 minor (41 kd and 12 kd) polypeptides. Preliminary evidence suggests a glycoprotein of 76 kd (GP 76) and a major doublet polypeptide of 13.5/13 kd to be also of viral origin. Of these GP 76 was susceptible to bromelain action implying its surface location in the virion, while p 32, p 26 and p 13.5/13 were the main constituents of viral cores. p 13.5/13 bound an RNA probe, suggesting it to be the main constituent of viral ribonucleoprotein. p 22/21 was not cleaved by bromelain, and was absent in viral cores suggesting its intramembrane location between virion envelope and core. The polypeptide profile of LPDV is distinct from those of avian sarcoma-leukosis viruses and avian reticuloendotheliosis viruses. PMID- 3038052 TI - Variation in the responses of Culicoides variipennis (Diptera, Ceratopogonidae) to oral infection with bluetongue virus. AB - The infection rate of an established colony of Culicoides variipennis to oral infection with bluetongue virus (BTV) type 4 was found to be highly variable and ranged from 0 to 51.6 per cent. This type of wide variation has previously only been reported to occur between individual populations of a species and not between samples from a single population. This must be taken into account when such colonies are used as reference standards for interpretation of field and laboratory data, particularly relating to studies of vector efficiency. Study of the response of individual susceptible C. variipennis to BTV infection demonstrated considerable variation in the level of virus multiplication that individual females are able to support. Virus concentrations varied from less than 1.1 to 5.1 log10 TCID50/fly up to 10 days post infection. 43.6 per cent of females contained less than 2.5 log10 TCID50 virus. It is suggested that such insects would have a mesenteron escape barrier to infection and would be incapable of transmitting BTV. The implications of these results for vector research are discussed. PMID- 3038053 TI - The pathogenesis of herpetic ocular disease in the guinea pig. AB - A detailed study of the pathogenesis of herpetic eye disease in the guinea pig was undertaken to further develop this animal model. Several well-known HSV-1 strains were tested for their ability to produce disease and cause acute and latent infections of the trigeminal ganglion: McKrae, KOS, McIntyre, RE, and Shealey. Two HSV-2 strains failed to cause eye infections. The Shealey strain [HSV-1 (Sh)] produced the most severe eye infections, characterized by epithelial and stromal disease, corneal vascularization and ulcerative blepharitis. Consequently, HSV-1 (Sh) was selected as the prototype strain for this study. The frequency and severity of HSV-1 (Sh) eye disease patterns was determined by a semi-quantitative rating scale, which permitted accurate monitoring of the temporal development of the disease patterns cited above. Virus shedding from infected eyes was also quantified. All of the HSV-1 strains tested established trigeminal ganglionic latency with varying frequency, although HSV-1 (Sh) latency approached 100 percent. The kinetics of acute ganglionic infection by HSV-1 (Sh) was determined, and peak virus titers occurred on the third day after corneal inoculation. This study emphasizes the usefulness of the Guinea pig model for investigations on the pathogenesis, prevention and treatment of herpetic eye infections. PMID- 3038054 TI - The polypeptides of human parainfluenza type 2 virus and their synthesis in infected cells. AB - We have studied the structural components of three strains of human parainfluenza virus type 2 (HPI-2) and identified the virus-specific polypeptides. Molecular weight of P and F1 polypeptides determined by us, when compared with that reported previously, was in reversed order. HN polypeptide existed chiefly as disulfide-linked dimers in cells infected with Toshiba strain, while as monomers and dimers in nearly equal proportion in cells infected with two other strains. Similar disulfide-linked NP oligomers were found in cells infected with all three strains. F1 and F2, cleaved forms of F protein, could be detected in cells infected with all three strains, but the ratio of cleaved (F1 and F2) to uncleaved (F0) forms was markedly lower in 62-M 786- and 63-M 1-infected than in Toshiba strain-infected cells. However, there was no difference of oligopeptide mapping patterns and isoelectric point of F polypeptide between Toshiba and 62-M 786 strains. By contrast, oligopeptide mapping patterns of HN protein of Toshiba strain differed from those of the two other strains. Furthermore, the HN polypeptide of Toshiba strain was phosphorylated in the infected Vero cells, but that of the other two strains was not. PMID- 3038055 TI - Transformation of rabbit arterial smooth muscle cells with simian virus 40. AB - In vitro studies were carried out to induce viral transformation of vascular smooth muscle cells. Cultured rabbit arterial smooth muscle cells were infected with simian virus 40 (SV 40), and transformed cultures were produced that exhibit altered morphology, increased growth rate and plating efficiency, growth on semi solid substrate, and chromosomal abnormalities. Nuclear SV 40 T-antigen was detected in all cells of these cultures. Muscle-specific actin was identified by a specific monoclonal antibody suggesting retention of smooth muscle cell characteristics by the transformed cells. Significant cytoplasmic lipid accumulation occurred in transformed cells incubated with beta-very low density lipoprotein, as revealed both by chemical analyses and Nile Red lipid staining of the culture. The transformed smooth muscle cells grow permanently in cell culture. Our investigations show that arterial smooth muscle cells transformed with SV 40 virus exhibit altered phenotypic properties distinct from that of normal arterial smooth muscle cells. PMID- 3038056 TI - The pathogenesis of rat virus infection in infant and juvenile rats after oronasal inoculation. AB - The pathogenesis of rat virus (RV) infection was studied in random-bred Sprague Dawley rats after oronasal inoculation of a recent RV isolate designated RV-Yale (RV-Y). RV-Y was pathogenic for rats inoculated as infants (2 days) whereas rats inoculated as juveniles (4 weeks) had asymptomatic infection and no lesions. Rats inoculated as infants developed pantropic infection accompanied by hepatic necrosis, granuloprival cerebellar hypoplasia and hemorrhagic encephalopathy. Virological and serological studies showed that virus could persist in inoculated rats for at least 35 days and for at least 28 days after seroconversion was first detected. Immunohistochemical results indicated that RV-Y infects tissues conducive to virus excretion including kidney and lung. RV-Y also was found in genital tissues of some rats. Athymic juvenile rats inoculated intraperitoneally with RV-Y had a poor humoral immune response and harbored infectious virus for at least 3 weeks, whereas infection in euthymic control rats was detected for 1 week. These studies indicate that RV-Y can persists in the presence of humoral immunity and suggest that transmission of infection could occur for a substantial period after seroconversion. They also suggest that immunodeficient rats have increased susceptibility to persistent infection. PMID- 3038057 TI - [Clinico-morphological characteristics of a juvenile angiofibroma of the base of the skull]. AB - 203 patients with juvenile angiofibroma of the base of the skull were assessed clinicoanatomically. In addition to a routine otorhinolaryngological examination 60 patients underwent computed tomography, radioangiography and thermography. Biopsy specimens obtained at operations were studied morphologically for 133 patients including application of histotopography in 15 cases and electron microscopy in 19 cases. Two clinicoanatomic forms of juvenile angiofibroma (JAF) have been revealed as well as 4 vascular types and variegation due to different combinations of vascular, cellular and fibrillar components. According to biologic nature of JAF it should be referred to fibromatoses. PMID- 3038058 TI - [Abrikosov's tumor of the lobular bronchus]. AB - The authors present a brief review of the literature on Abrikosov's tumour, as well as their own observation. In a patient, aged 32, the X-ray examination revealed a supralobar bronchus tumour. Bronchoscopically the tumour occluded the bronchial lumen. Left lobectomy was performed. Histologically Abrikosov's tumour arose from the submucosal bronchial layer and consisted of large spherical and polygonal cells with a light eosinophilic granular cytoplasm. PMID- 3038059 TI - [Herpetic brain stem encephalitis: report of an autopsied case with immunohistochemical and electron microscopy studies]. AB - A post-mortem examined case of herpetic brainstem encephalitis is presented. Clinically, the patient had cephalalgia followed by ataxia, drowsiness and multiple palsies of some cranial nerves, developing into death in eight days. The pathologic examination of the brain showed necrotizing encephalitis in multiple foci limited to the brainstem, more distinctly in the pons and medulla oblongata. The technique of immunoperoxidase revealed rare glial cells with intranuclear immunoreactivity for herpes antigen. Rare viral particles with the morphological characteristics of the herpesvirus were identified in the nuclei of neurons in 10% formol fixed material. This is the second reported case of herpetic brainstem encephalitis confirmed by post-mortem examination. The pathway used by the virus to reach the central nervous system and its posterior dissemination to the oral cavity, the orbitofrontal region and the temporal lobes as well as to the brainstem, after a period of latency and reactivation, are discussed. PMID- 3038060 TI - Malignant fibrous histiocytoma of the orbit in a 3-year-old girl. Case report. PMID- 3038061 TI - [Immunopathology of the upper respiratory tract]. PMID- 3038062 TI - Downregulation of high density lipoprotein receptor activity of cultured fibroblasts by platelet-derived growth factor. AB - High density lipoprotein (HDL) receptor activity was decreased by the addition of platelet-derived growth factor (PDGF) to cholesterol-loaded cultured human skin fibroblasts. Effects were observed within 12 hours, coinciding with increases in tritiated thymidine incorporation into DNA. The PDGF-mediated decrease in HDL binding was associated with a decrease in cholesterol efflux promoted by HDL3. PDGF exerted reciprocal effects on HDL and low density lipoprotein (LDL) receptor activity. With fibroblasts not loaded with cholesterol, PDGF increased LDL binding without changing HDL receptor activity. With cholesterol-loaded cells, PDGF decreased HDL receptor activity without changing LDL receptor activity. These results show that a reduction in HDL receptor activity is an additional cellular response to PDGF. Decreased HDL-mediated cholesterol efflux may result in retention of cholesterol needed for membrane synthesis in cells stimulated to proliferate. PMID- 3038063 TI - Arsenic poisoning and peripheral neuropathy. PMID- 3038064 TI - Congestive heart failure. Pathophysiology and treatment. Part Three. PMID- 3038065 TI - Bile salt-induced injury of rabbit oesophageal mucosa measured by hydrogen ion disappearance. AB - Oesophageal injury secondary to gastro-oesophageal reflux is unlikely to be due to the effects of hydrochloric acid alone. The present authors have investigated the development of acid and bile salt-induced oesophageal mucosal injury in a rabbit model. Solutions of hydrochloric acid and sodium taurocholate (ST) were perfused through an isolated oesophageal preparation and mucosal injury was determined by measuring the rate of H+ disappearance. Perfusion with acid alone in concentrations up to 10 mmol/l did not affect the H+ disappearance rate. Addition of 1 mmol/l ST to an acid perfusate resulted in loss of H+ from the system. The increase in H+ disappearance rate was associated with loss of ST from the perfusate. Sodium taurocholate was only lost from the system when in an acid medium. Increased rate of H+ disappearance occurred even after the bile salt had been washed out of the perfused oesophagus. The mechanism of bile salt-induced mucosal injury was unlikely to be due to mucosal disruption secondary to micelle formation since the critical micellar concentration of taurocholate was found to be greater than that used in the perfusate. These findings indicate that bile salts may be an important factor in hydrochloric acid-related damage to oesophageal mucosa, by acting through mechanisms unrelated to micelle formation. PMID- 3038066 TI - Oestradiol binding activity in rat liver tumours and in human hepatocellular carcinoma. AB - Oestradiol binding activity was studied in normal liver tissues of rats as well as in experimentally induced liver tumours by incubating a cytosol prepared from the tissue with radioactive oestradiol. The free radioactive hormone was absorbed by a dextran-charcoal suspension, and the amount required to saturate the receptors was calculated by a Scatchard plot. Similar activity was observed in normal human liver as well as naturally occurring hepatocellular carcinoma. PMID- 3038067 TI - The experimental infection of horses with Murray Valley encephalitis and Ross River viruses. AB - Eleven weanling horses were inoculated with Murray Valley encephalitis and Ross River viruses either by intravenous injection or by the bite of Culex annulirostris or Aedes vigilax mosquitoes infected orally. Five of the 11 horses circulated trace amounts of MVE virus for 1 to 5d and they infected 7/408 Cx annulirostris which subsequently fed on them. Haemagglutination-inhibiting antibody persisted at detectable levels for the 24-week observation period. With Ross River virus, only one of 11 horses inoculated developed a viraemia detectable by inoculation of suckling mice but 5 horses contained virus sufficient to infect 41/383 Cx annulirostris that fed on them 3 to 4 days after inoculation. On primary inoculation with Ross River virus, only 2 horses developed HI antibody but late responses occurred in 3 horses following probable naturally acquired re-infections. With both viruses, most horses remained normal, some developed mild pyrexia and transient clinical signs. This paper, therefore, indicates that horses are unlikely to be efficient amplifiers of either MVE or RR viruses and does little to incriminate them as important pathogens. PMID- 3038068 TI - Hippocampal associative cellular responses: dissociation with behavioral responses revealed by a transfer-of-control technique. AB - Multiunit activity was recorded in the CA3 field of the dorsal hippocampus in freely moving rats during classical conditioning and subsequent presentation of the CS on operant baselines for food reward as well as shock avoidance. Rats were first trained in a nonsignaled bar-pressing-dependent shock omission task and in a food-motivated lever-pressing task (60-s VI). Five sessions with presentations of a previously habituated tone as a CS paired with footshock as a US were then given. Testing was carried out by presenting the CS alone while behavioral responses were maintained by reinforcement in both instrumental tasks on alternate sessions. As expected, the CS induced a marked suppression of lever pressing for food reward and a marked enhancement of bar-pressing for shock avoidance. The analysis of the frequency of multiunit discharges to the CS revealed that the hippocampal cellular responses established during classical conditioning were maintained while two different behavioral responses were exhibited to the CS. The results showed that the associative response of hippocampal neurons may be dissociated from the Pavlovian conditioned responses the CS elicits. They support the hypothesis that hippocampal cellular responses represent a neural index of the acquired CS-US associative representation. PMID- 3038069 TI - The role of lipid peroxidation in pathogenesis of arrhythmias and prevention of cardiac fibrillation with antioxidants. AB - The present paper shows the arrhythmogenic effect of a direct induction of lipid peroxidation (LP) on isolated auricles; it is demonstrated that preendured stress potentiates this effect, while antioxidants prevent it. Subsequently, in studying the mechanism of the LP arrhythmogenic effect it was established that stress, like the LP induction, disorders the activity of Na, K-ATPase and accelerates thermodenaturation of this enzyme which plays a key role in maintaining the transmembrane potential and the electrical stability of the heart. Antioxidants prevent the enumerated shifts. Based on these data, the antioxidant BHT was successfully applied for prevention of the fall in cardiac fibrillation threshold in stress and experimental myocardial infarction, and also for prevention of cardiac fibrillation itself under acute ischemia and reoxygenation of the heart. PMID- 3038070 TI - Vasoactive effects of eicosapentaenoic acid on isolated vascular smooth muscle. AB - Dietary intake of unsaturated fatty acid of eicosapentaenoic acid (EPA) is thought to reduce the size and incidence of myocardial infarction. These beneficial effects are postulated to be due to chronic antithrombotic properties of EPA itself. We studied the possible direct effects of EPA on vascular smooth muscle as well as the ability of EPA to modify the vasoactivity of constrictor mediators in rabbit and cat aortic rings and isolated cat coronary arteries. EPA concentration-dependently (30 to 300 microM) relaxed rabbit and cat aortic rings having an intact endothelium, while EPA did not show any significant vasodilator effects on rings without an endothelium. This EPA-induced vasorelaxation was not altered by the cyclooxygenase inhibitor ibuprofen, but was totally abolished by the guanylate cyclase inhibitor methylene blue, indicating an endothelium dependent smooth muscle relaxation mechanism. In isolated perfused cat coronary arteries, EPA (3 to 300 microM) exerted a dilator effect which was endothelium independent and not affected by ibuprofen. The response was attenuated by propyl gallate, a lipoxygenase inhibitor. EPA also inhibited leukotriene (LT) C4, (50 nM) and LTD4 (50 nM)-induced vasoconstriction of isolated cat coronary arteries ranging from a blockade of 10% to 15% (P less than 0.05) at 3 microM of EPA to a blockade of 89% to 93% (P less than 0.01) at 300 microM. In contrast, the thromboxane analog, CTA2, induced coronary constriction was not significantly altered by EPA. Thus, EPA produces endothelium-dependent relaxation in rabbit and cat aorta and endothelium-independent vasodilation in cat coronary arteries (i.e., intact vessels or helical strips). Moreover, EPA exerts acute anti leukotriene actions in coronary arteries. In the case of long-term dietary intake of EPA, these actions may contribute to the protective action of EPA in myocardial ischemia. PMID- 3038071 TI - [Equine herpesvirus 1 (EHV-1) infection in the horse: neurologic symptoms in a standard bred mare with acute fatal course. Molecular characterization of the brain isolates and pathologic correlates]. PMID- 3038072 TI - [Comparative determinations of antibodies following vaccination of swine and guinea pigs with inactivated swine parvovirus vaccines]. PMID- 3038073 TI - Dietary (n-3) fatty acids and eicosanoid formation. PMID- 3038074 TI - Localization of the inosine triphosphatase locus (Itp) on chromosome 2 of the mouse. AB - An electrophoretic variant of the enzyme inosine triphosphatase was found by screening inbred strains of mice. Strains with the slower-migrating variant include BALB/cJ, DBA/1J, and PL/J. The Itp locus was mapped between the beta-2 microglobulin (B2m) and the agouti (a) loci on chromosome 2. The mapping of Itp on chromosome 2 identifies a chromosomal segment that has been conserved since the divergence of lineages leading to mouse and man. PMID- 3038076 TI - Pseudomonas mutant strains that accumulate androstane and seco-androstane intermediates from bile acids. AB - Transposon mutant strains which were affected in bile acid catabolism were isolated from four Pseudomonas spp. Two of the mutant groups isolated were found to accumulate 12 alpha-hydroxyandrosta-1,4-diene-3,17-dione as the major product from deoxycholic acid. Strains in one of these two groups were able to grow on steroids such as chenodeoxycholic acid, which lacks a 12 alpha-hydroxy function, whereas the one member of the second group could not. With chenodeoxycholic acid, this latter strain accumulated a yellow muconic-like derivative, tentatively identified as 3,7-dihydroxy-5,9,17-trioxo-4(5),9(10)-disecoandrosta-1(10)2 -dien 4-oic acid. Members of two further mutant groups accumulated either 12 beta hydroxyandrosta-1,4-diene-3,17-dione or 3,12 beta-dihydroxy-9(10)-secoandrosta 1,3,5(10)-triene-9,17-dione as the major product from deoxycholic acid. The relationship between the catabolism of m- and p-cresol, 3-ethylphenol and the bile acids was also examined. PMID- 3038077 TI - Conformational equilibria of alpha-L-iduronate residues in disaccharides derived from heparin. AB - The disaccharides IdoA(2SO3)-anManOH(6SO3) and IdoA-anManOH (where IdoA represents alpha-L-iduronate, anManOH represents 2,5-anhydro-D-mannitol and SO3 represents sulphate ester) were prepared from bovine lung heparin using HNO2 depolymerization, borohydride reduction and desulphation, and were examined by 400 MHz 1H-n.m.r. spectroscopy. Three-bond proton-proton coupling constants around the IdoA ring were determined under a range of experimental conditions. For unsulphated IdoA all four proton-proton coupling constants varied markedly as a function of temperature, pH and solvent, providing clear evidence for a rapid conformational equilibrium. These data were analysed in terms of the three most energetically stable IdoA conformers: 1C4, 4C1, and 2S0. Predicted coupling constants for these conformers were determined using a modified Karplus-type relationship. For unsulphated IdoA in dimethyl sulphoxide the equilibrium was provoked strongly in favour of a slightly distorted 4C1 'chair' IdoA conformer for which coupling constants have not previously been reported. For sulphated IdoA in aqueous conditions and at low pH the equilibrium is strongly in favour of the alternative 1C4 chair conformer. Under many conditions, however, significant contributions from all three conformers occur for the non-reducing terminal IdoA in these disaccharides. PMID- 3038075 TI - DNA structure and perturbation by drug binding. PMID- 3038078 TI - The hydrolysis of alpha-human atrial natriuretic peptide by pig kidney microvillar membranes is initiated by endopeptidase-24.11. AB - alpha-Human atrial natriuretic peptide, a 28-amino-acid-residue peptide, was rapidly hydrolysed by pig kidney microvillar membranes in vitro, with a t1/2 of 8 min, comparable with the rate observed with angiotensins II and III. The products of hydrolysis were analysed by h.p.l.c., the pattern obtained with membranes being similar to that with purified endopeptidase-24.11 (EC 3.4.24.11). No hydrolysis by peptidyl dipeptidase A (angiotensin I converting enzyme, EC 3.4.15.1) was observed. The contribution of the various microvillar membrane peptidases was assessed by including specific inhibitors. Phosphoramidon, an inhibitor of endopeptidase-24.11, caused 80-100% suppression of the products. Captopril and amastatin (inhibitors of peptidyl dipeptidase A and aminopeptidases respectively) had no significant effect. Hydrolysis at an undefined site within the disulphide-linked ring occurred rapidly, followed by hydrolysis at other sites, including the Ser25--Phe26 bond. PMID- 3038079 TI - Inositol 1,2-cyclic 4,5-trisphosphate is not a product of muscarinic receptor stimulated phosphatidylinositol 4,5-bisphosphate hydrolysis in rat parotid glands. AB - We have employed a neutral-pH extraction technique to look for inositol 1,2 cyclic phosphate derivatives in [3H]inositol-labelled parotid gland slices stimulated with carbachol. The incubations were terminated by adding cold chloroform/methanol (1:2, v/v), the samples were dried under vacuum and inositol phosphates were extracted from the dried residues by phenol/chloroform/water partitioning. Water-soluble inositol metabolites were separated by h.p.l.c. at pH 3.7. 32P-labelled inositol phosphate standards (inositol 1-phosphate, inositol 1,2-cyclic phosphate, inositol 1,4,5-trisphosphate and inositol 1,2-cyclic 4,5 trisphosphate) were quantitively recovered through both extraction and chromatography steps. Treatment of inositol cyclic phosphate standards with 5% (w/v) HClO4 for 10 min prior to chromatography resulted in formation of the expected non-cyclic compounds. [3H]Inositol 1-phosphate and [3H]inositol 1,4,5 trisphosphate were both present in parotid gland slices and both increased during stimulation with 1 mM-carbachol. There was no evidence for significant quantities of [3H]inositol 1,2-cyclic phosphate or [3H]inositol 1,2-cyclic 4,5-trisphosphate in control or carbachol-stimulated glands. Parotid gland homogenates rapidly converted inositol 1,4,5-trisphosphate to inositol bisphosphate and inositol tetrakisphosphate, but metabolism of the inositol cyclic trisphosphate was much slower. The results suggest that inositol 1,4,5-trisphosphate, but not inositol 1,2-cyclic 4,5-trisphosphate, is the water-soluble product of muscarinic receptor stimulated phospholipase C in rat parotid glands. PMID- 3038080 TI - The antioxidant action of human extracellular fluids. Effect of human serum and its protein components on the inactivation of alpha 1-antiproteinase by hypochlorous acid and by hydrogen peroxide. AB - The elastase-inhibitory capacity of purified human alpha 1-antiproteinase is inactivated by low concentrations of the myeloperoxidase-derived oxidant hypochlorous acid, but much higher concentrations are required to inhibit the elastase-inhibitory capacity of serum samples. The protective effect of serum appears to be largely due to albumin. High concentrations of H2O2 also inactivate the elastase-inhibitory capacity of alpha 1-antiproteinase, by a mechanism not involving formation of hydroxyl radicals. Serum offers protection against H2O2 inactivation of alpha 1-antiproteinase. The relevance of these results to the tissue damage produced by activated phagocytes is discussed. PMID- 3038081 TI - Purification and properties of formate dehydrogenase from Pseudomonas aeruginosa. Characterization of haem and iron-sulphur centres by magnetic-circular-dichroism and electron-paramagnetic-resonance spectroscopy. AB - The purification of formate dehydrogenase (FDH) from Pseudomonas aeruginosa after anaerobic growth on nitrate-containing medium was carried out. The separation of the FDH enzyme from nitrate reductase (NiR), which are found together in a particle fraction and constitute the short respiratory chain of this bacterium, has been followed by optical, magnetic c.d. (m.c.d.) and e.p.r. spectroscopy. These techniques have allowed the haem, iron-sulphur clusters and molybdenum components to be detected and, in part, their nature to be determined. Attempts to extract FDH anaerobically in the absence of sodium dithionite led to loss of activity. Addition of sodium dithionite maintained the activity of the enzyme, even after subsequent exposure to air, in an assay involving formate reduction with Nitro Blue Tetrazolium as reductant. Three preparations of FDH have been examined spectroscopically. The preparations vary in the amount of contaminating nitrate reductase, the amount of cytochrome c present and the concentration of oxidized [3Fe-4S] cluster. Optical spectra and low-temperature m.c.d. spectroscopy show the loss of a cytochrome-containing protohaem IX co-ordinated by methionine and histidine as NiR is separated from the preparation. In its purest state FDH contains one molecule of cytochrome co-ordinated by two histidine ligands in the oxidized state. This cytochrome has an e.p.r. spectrum with gz = 3.77, the band having the unusual ramp shape characteristic of highly anisotropic low-spin ferric haem. It also shows a charge-transfer band of high intensity in the m.c.d. spectrum at 1545 nm. It has recently been shown [Gadsby & Thomson (1986) FEBS Lett. 197, 253-257] that these spectroscopic properties are diagnostic of a bishistidine co-ordinated haem with steric constraint of the axial ligands. The e.p.r. and m.c.d. spectra of the reduced state of FDH reveal the presence of an iron-sulphur cluster of the [4Fe-4S]+ type. The g-values are 2.044, 1.943 and 1.903. An iron-sulphur cluster of the class [3Fe-4S], detected by e.p.r. spectroscopy in the oxidized state and by low-temperature m.c.d. spectroscopy in the reduced state, is purified away with the NiR. Finally, an e.p.r. signal at g = 2.0 with a narrow bandwidth which persists to 80 K is observed in the purest preparation of FDH. This may arise from an organic radical species. PMID- 3038082 TI - Purification and properties of formate dehydrogenase from Pseudomonas aeruginosa. Electron-paramagnetic-resonance studies on the molybdenum centre. AB - Formate dehydrogenase from Pseudomonas aeruginosa contains molybdenum, a [4Fe-4S] cluster and cytochrome b. This paper reports the detection of molybdenum as Mo(V) by e.p.r. spectroscopy. In order to generate Mo(V) signals, addition of amounts of excess formate varying between 10- and 50-fold over enzyme, followed by 200 fold excess of sodium dithionite, were used. Two Mo(V) species were observed. One, the major component, has g1 = 2.012, g2 = 1.985 and g3 = 1.968, appeared at low concentrations of formate and increased linearly in intensity with increasing concentrations of formate up to 25-fold excess over the enzyme. At higher formate concentration this signal disappeared. The appearance and disappearance of this Mo(V) signal seems to parallel the state of reduction of the [4Fe-4S] clusters. A second, minor, Mo(V) species with g-values g1 = 1.996, g2 = 1.981 and g3 = 1.941 appears at a constant level during the formate-dithionite titration. No evidence has been obtained for nuclear hyperfine coupling to protons. The major Mo(V) species has unusual e.p.r. signals compared with other molybdenum-containing enzymes, except for that observed in the formate dehydrogenase from Methanobacterium formicicum [Barber, Siegel, Schauer, May & Ferry (1983) J. Biol. Chem. 258, 10839-10845]. The present work suggests that the enzyme is acting as a CO2 reductase, with dithionite as an electron donor to a [4Fe-4S] cluster, which in turn donates electrons to molybdenum, producing a Mo(V) species with CO2 bound to the metal. PMID- 3038083 TI - Electron-paramagnetic-resonance and magnetic-circular-dichroism studies on the formate dehydrogenase-nitrate reductase particle from Pseudomonas aeruginosa. AB - The membrane-bound respiratory particle complex of Pseudomonas aeruginosa, which reduces nitrate to nitrite using formate as the electron donor, was prepared and characterized by e.p.r. and low-temperature magnetic c.d. (m.c.d.) spectroscopy. The particle complex has two enzymic components, namely nitrate reductase (NiR) and formate dehydrogenase (FDH), which are multi-centred proteins containing molybdenum, iron-sulphur clusters and cytochrome. By using results from work on the purified extracted enzymes NiR and FDH to aid in the assignment, it has been possible to observe spectroscopically all the components of the electron-transfer chain in the intact particle. This led to a proposal for the organization of the metal components of the FDH-NiR chain. Molybdenum ions are at opposite ends of the chain and interact with, respectively, the formate-CO2 couple and the nitrate nitrite couple. The molybdenum ion at the low-potential end of the chain passes electrons to cytochrome b of FDH, a bishistidine-co-ordinated haem with unusual steric restraint at the iron. The next component is a [4Fe-4S] cluster. This comprises all the components of FDH. Electrons are passed to the molybdenum of NiR via a number, probably two, of [4Fe-4S] clusters. No evidence has been found in this work for the presence of a quinone to mediate electron transfer between FDH and NiR. Cytochrome c appears to be able to feed electrons into the chain at the level of one of the [4Fe-4S] centres of NiR. PMID- 3038084 TI - The bisphosphonomethyl analogue of 2,3-bisphosphoglycerate inhibits yeast but not wheat-germ phosphoglycerate mutase. AB - The bisphosphonomethyl analogue of 2,3-bisphosphoglycerate [4-phosphono-2 (phosphonomethyl) butanoate] was a potent competitive inhibitor of cofactor dependent phosphoglycerate mutase from yeast, with a Ki of 0.8 mM. In contrast, the analogue did not affect the activity of cofactor-independent phosphoglycerate mutase from wheat germ. It is considered that this compound should be particularly useful for n.m.r. spectroscopic studies on the mechanism of action of cofactor-dependent phosphoglycerate mutases. PMID- 3038085 TI - The rapid desensitization of glucagon-stimulated adenylate cyclase is a cyclic AMP-independent process that can be mimicked by hormones which stimulate inositol phospholipid metabolism. AB - Treatment of intact hepatocytes with glucagon, TH-glucagon [( 1-N-alpha trinitrophenylhistidine, 12-homoarginine]glucagon), angiotensin or vasopressin led to a rapid time- and dose-dependent loss of the glucagon-stimulated response of the adenylate cyclase activity seen in membrane fractions isolated from these cells. Intracellular cyclic AMP concentrations were only elevated with glucagon. All ligands were capable of causing both desensitization/loss of glucagon stimulated adenylate cyclase activity and stimulation of inositol phospholipid metabolism in the intact hepatocytes. Maximally effective doses of angiotensin precluded any further inhibition/desensitizing action when either glucagon or TH glucagon was subsequently added to these intact cells, as has been shown previously for the phorbol ester TPA (12-O-tetradecanoylphorbol 13-acetate) [Heyworth, Wilson, Gawler & Houslay (1985) FEBS Lett. 187, 196-200]. Treatment of intact hepatocytes with these various ligands caused a selective loss of the glucagon-stimulated adenylate cyclase activity in a washed membrane fraction and did not alter the basal, GTP-, NaF- and forskolin-stimulated responses. Angiotensin failed to inhibit glucagon-stimulated adenylate cyclase activity when added directly to a washed membrane fraction from control cells. Glucagon GR2 receptor-stimulated adenylate cyclase is suggested to undergo desensitization/uncoupling through a cyclic AMP-independent process, which involves the stimulation of inositol phospholipid metabolism by glucagon acting through GR1 receptors. This action can be mimicked by other hormones which act on the liver to stimulate inositol phospholipid metabolism. As the phorbol ester TPA also mimics this process, it is proposed that protein kinase C activation plays a pivotal role in the molecular mechanism of desensitization of glucagon-stimulated adenylate cyclase. The site of the lesion in desensitization is shown to be at the level of coupling between the glucagon receptor and the stimulatory guanine nucleotide regulatory protein Gs, and it is suggested that one or both of these components may provide a target for phosphorylation by protein kinase C. PMID- 3038086 TI - Release of iron from ferritin by xanthine oxidase. Role of the superoxide radical. AB - Mobilization of iron from ferritin by xanthine oxidase was studied under aerobic and anaerobic conditions. Aerobic iron release amounted to approx. 3.7 nmol/ml in 10 min. This amount was decreased by approx. 30% under anaerobic conditions. Aerobic iron mobilization involved two mechanisms. About 70% was released by O2.- generated by xanthine oxidase. The rest was released by O2(.-)-independent mechanisms, which also accounted for the total iron release when O2 was absent. A possible transfer of reducing equivalents directly from xanthine oxidase to ferritin is discussed. The results imply that, in pathological conditions with increased formation of O2.-, iron may be released from ferritin. Furthermore, in hypoxic tissues xanthine oxidase can release iron from ferritin by an O2(.-) independent process. Free iron is liable to catalyse the formation of the extremely reactive and damaging OH. radical. PMID- 3038087 TI - The effect of disodium ethane-1-hydroxy-1,1-diphosphonate on the metabolism of calcitriol in chicks. AB - Decreased intestinal absorption of Ca2+ occurs in response to treatment with disodium ethane-1-hydroxy-1,1-diphosphonate (EHDP). The effect is due to decreased 1-hydroxylation of calcidiol (25-hydroxycholecalciferol) in the kidney. In an attempt to establish whether impairment of vitamin D metabolism at steps beyond kidney hydroxylation occurs due to treatment with EHDP, chicks were depleted of vitamin D and were treated with calcitriol (1,25 dihydroxycholecalciferol) as their sole source of the vitamin. The chicks were then divided into two groups, one being treated with EHDP while the second group served as control. Intestinal absorption of Ca2+ in the EHDP-treated group was found to be impaired, along with decreases in concentrations of calbindin D28K (the 28,000-Mr vitamin D-dependent Ca2+-binding protein). When the chicks were dosed with [3H]calcitriol, significantly lower concentrations of the sterol were detected in the duodena of EHDP-treated birds. Measurement of levels of receptors for calcitriol in duodena showed no difference between groups, but levels of calcitriol in sera were considerably lower in the EHDP-treated group along with the elevated biliary and urinary excretion of glucuronidated conjugates. It is therefore concluded that treatment with EHDP results in increased catabolism of calcitriol in addition to the known suppression of the renal production of the hormone. PMID- 3038088 TI - Inactivation of angiotensin converting enzyme by sodium nitroprusside. AB - Angiotensin I converting enzyme is rapidly inactivated by sodium nitroprusside and that inactivation is suppressed in the presence of chloride ion and by the presence of L-alanyl-L-proline or glycyl-L-tryptophan, which are both competitive inhibitors of its catalytic activity. The inactivation by sodium nitroprusside appears to result from the modification of an unusually reactive lysine residue in or near the active site. PMID- 3038089 TI - Segregation of nucleosomes in replicated mouse alpha-globin gene. AB - DNA of mouse erythroleukemia cells grown in vitro was labeled with bromodeoxyuridine during cycloheximide-inhibited protein synthesis. Isolated nuclei were digested with micrococcal nuclease to obtain monosomes and monosomal dsDNA. The protection of the heavy and of the light strands of the newly replicated DNA was studied by dot hybridization with the coding and with its complementary noncoding strand of the alpha-globin gene. The results show that both sides of the replication fork contain protected sequences of the gene, thus supporting a bilateral (dispersive) mode of nucleosome segregation during DNA replication. PMID- 3038090 TI - Neurotensin receptors on the rat liver plasma membranes. AB - Neurotensin (NT) is now classified as a brain-gut peptide in the central nervous system and gastrointestinal tract. In the present study, we characterized the NT receptors on the rat liver plasma membranes. The specific binding of [3H]NT was time dependent, reversible, and saturable. Scatchard analysis of the specific binding data yielded two classes of binding sites, a high affinity site and a low affinity site. The average maximum number of binding sites (Bmax) amounted to 13.3 +/- 1.1 fmol/mg protein at high affinity site and 122.3 +/- 21.5 fmol/mg protein at low affinity site, respectively. The dissociation constant (Kd) had values of 0.39 +/- 0.01 nM at high affinity site and 8.1 +/- 1.1 nM at low affinity site, respectively. The amount of specifically bound [3H]NT was significantly reduced in the presence of mono and divalent cations, EDTA, EGTA and a peptidase inhibitor bacitracin, NT1-13 competed with [3H]NT for its binding site with an IC50 of 0.19 nM at high affinity site (0.2 nM concentration of [3H]NT) and 0.7 nM at low affinity site (4.0 nM concentration of [3H]NT). Xenopsin, a NT analogue separated from the skin of Xenopus laevis, was equipotent (IC50 0.75 nM) with NT1-13 at 4.0 nM concentration of [3H]NT. C-terminal sequence of NT contains the structure necessary for interaction with NT binding sites whereas N-terminal sequence had no binding activity. Since NT has a hyperglysemic and a hypercholesterolemic effects in rats, these NT receptors on the rat liver plasma membranes may be involved in the hyperglycemia and/or hypercholesteroremia induced by NT. PMID- 3038091 TI - 1H-NMR investigation of yeast cytochrome c. Interaction with the corresponding specific reductase (L-lactate cytochrome). AB - 1H-NMR spectroscopy has been used to study the modifications of certain characteristic resonances of the Hansenula anomala yeast cytochrome c on binding to its specific reductase (flavocytochrome b2) or to the isolated cytochrome domain obtained from the entire molecule. Normal titration curves are observed for the resonances at 37.8 ppm assigned to heme c methyl 8 and at 19.4 ppm, line of cytochrome b2 spectrum. In contrast, the shifts near 3.2 and 3.4 ppm for trimethyl-lysine resonances of this cytochrome c present abnormal titration curves, saturation being apparently reached at low molar (cytochrome b2)/(cytochrome c) ratio. An interpretation is proposed in terms of shifts due to local conformational transitions induced by reductase binding but not rapidly reversible upon dissociation. PMID- 3038092 TI - Cell-free activation of phagocyte NADPH-oxidase: tissue and differentiation specific expression of cytosolic cofactor activity. AB - We examined a variety of tissues for the presence of cytosolic cofactor activity that would support arachidonate-dependent cell-free activation of NADPH-oxidase in isolated human neutrophil membranes. Cofactor activity was not found in cytosol isolated from erythrocytes, lymphocytes, placenta, brain, liver, or the human promyelocytic leukemic cell line HL-60. Induction of differentiation in HL 60 cells led to expression of cytosolic cofactor activity. In dimethylsulphoxide induced HL-60 cells the level of cytosolic cofactor activity was closely correlated with phorbol myristate acetate-stimulated whole cell superoxide production. These results strongly suggest that the cytosolic cofactor is a phagocyte-specific regulatory protein of physiologic importance in NADPH-oxidase activation. PMID- 3038093 TI - Induction by fibroblast growth factor of angiotensin converting enzyme in vascular endothelial cells in vitro. AB - Induction of vascular endothelial cells with pituitary fibroblast growth factor (FGF) provoked an increase in angiotensin converting enzyme activity. The stimulatory effect of FGF on ACE activity was dose-dependent (ED50 = 1.0 ng/ml). Our results suggest a possible role for pituitary FGF in regulation of ACE production in vascular endothelial cells. PMID- 3038094 TI - Molecular cloning and sequence analysis of full-length cDNA for mRNA of adrenodoxin oxidoreductase from bovine adrenal cortex. AB - A full-length cDNA clone (pADR) for adrenodoxin reductase was isolated by means of immunological screening from a bovine adrenal poly(A)+ mRNA library. A cDNA insert of 1,973 base pairs in length encoded the entire amino acid sequence of the adrenodoxin reductase precursor protein, which consists of 492 amino acids including an extrapeptide of 32 amino acids at the NH2-terminus. The cloned cDNA contained the complete 3'-noncoding region of 443 nucleotides including 59 nucleotides of poly(A) and 51 nucleotides in the 5'-noncoding region. The amino acid sequences from the 33rd to 70th, the 117th to 123rd, the 207th to 225th, the 247th to 323rd, the 385th to 426th, the 444th to 461st, and the 487th to 492nd in the predicted structure were identical with those of the purified adrenodoxin reductase and its digested peptides, with only four exceptions. PMID- 3038095 TI - Decreased level of calpain inhibitor activity in kidney from Milan hypertensive rats. AB - Rat kidney contains two different calpain isozymes distinguishable on the basis of their Ca2+ requirement and of their activation mechanisms. The two calpain isozymes are present in comparable amounts in kidney of normotensive and hypertensive rats of the Milan strain. Conversely, the level of the natural inhibitor of calpain is significantly decreased in kidney of hypertensive rats as compared to control normotensive rats. This deficiency is more pronounced in the cortical region than in other kidney fractions. These results taken together with previous observations indicating the existence of an identical defect in red cells from the same hypertensive rat strain, (Pontremoli, S., Melloni, E., Salamino, F., Sparatore, B., Viotti, P., Michetti, M., Duzzi, L., and Bianchi, G. (1986) Biochem. Biophys. Res. Commun. 138, 1370-1375) emphasize the possible role of an unbalanced intracellular proteolytic system in the development of genetically determined hypertension. PMID- 3038096 TI - The Ca2+ -activated protease (calpain) modulates Ca2+/calmodulin dependent activity of smooth muscle myosin light chain kinase. AB - The Ca2+ -activated neutral protease can proteolyze both Ca2+ -dependent cyclic nucleotide phosphodiesterase and smooth muscle myosin light chain kinase. Ca2+ dependent cyclic nucleotide phosphodiesterase from rat brain was converted to the Ca2+ -independent active form by Ca2+ -activated protease. The proteolytic effects on myosin light chain kinase of Ca2+-activated protease differed in the presence and absence of the Ca2+-calmodulin (CaM) complex. In the presence of bound CaM, myosin light chain kinase (130k dalton) was degradated to a major fragment of 62 kDa, which had Ca2+/CaM-dependent enzyme and CaM-binding activity. When digestion occurred in the absence of bound CaM, myosin light chain kinase cleaved to a fragment of 60 kDa. This peptide had no enzymatic activity in the presence or absence of the Ca2+-CaM complex. Available evidence suggests that the Ca2+-activated proteases may recognize the conformational change of smooth muscle myosin light chain kinase induced by Ca2+-CaM complex. PMID- 3038097 TI - Increased phosphorylation in red cell membranes of subjects affected by essential hypertension. AB - In hemolysates of red cells from hypertensive patients the proteolytic activity of calpain is expressed at a rate approximately three fold higher than in red cells of normotensive subjects. Susceptibility to lysis upon exposure to ionophore A23187 and calcium, conditions that increase intracellular calpain activity, is also significantly enhanced in erythrocytes of hypertensive patients. In inside-out vesicles prepared from erythrocytes of these patients band 3 region undergoes a high extent of phosphorylation which is 1.5 fold higher than that occurring in control red cells from normotensive subjects. This increased phosphorylation can be reproduced in inside-out vesicles from erythrocytes of normal subjects following pretreatment with calpain. Taken together, these results suggest that the presence in erythrocytes of hypertensive subjects of an unregulated calpain dependent proteolytic activity may affect the structure of plasma membranes and determine an increased phosphorylation of intrinsic membrane proteins. PMID- 3038098 TI - Conversion of leukotriene B4 to dihydro and 19-hydroxy metabolites by rat polymorphonuclear leukocytes. AB - Rat polymorphonuclear leukocytes metabolize leukotriene B4 (LTB4) by at least two major pathways. LTB4 is converted by a reductase in these cells to a dihydro metabolite in which one of the three conjugated double bonds has been reduced to give a conjugated diene with a UV absorption maximum at 230 nm. DihydroLTB4 appears to be a key intermediate in the metabolism of LTB4 by rat polymorphonuclear leukocytes, since a number of other metabolites, exhibiting UV absorbance at 235 nm, but not at 280 nm, have been detected by high pressure liquid chromatography. In addition, these cells contain a 19-hydroxylase, which converts LTB4 to 19-hydroxyLTB4, which has a typical leukotriene UV spectrum, exhibiting absorption maxima at 261, 270, and 282 nm. PMID- 3038099 TI - Glucocorticoid induction of the maturation of ovine fetal adrenocortical cells. AB - The aim of the present study was to assess whether glucocorticoids could be directly involved in the maturation of adrenocortical cells from 120-138 days old ovine fetuses. The cAMP response to ACTH1-24 of cells cultured for 24 hours in the presence of ACTH1-24 was 2 fold higher than that of control cells. However, the response of cells cultured in the presence of ACTH1-24 plus metyrapone or aminoglutethimide was lower than that of cells cultured in the presence of ACTH1 24 alone. Cells cultured for 48 hours in the presence of dexamethasone or cortisol released more cAMP than control cells when stimulated by ACTH1-24, but not in response to forskolin. However corticosteroid production stimulated by ACTH1-24, forskolin or dibutyryl cAMP was enhanced by dexamethasone treatment. These results suggest that glucocorticoids can affect the maturation of ovine fetal adrenocortical cells by an auto and/or a paracrine process, and that this effect is exerted, at least, at two different levels in the cell. PMID- 3038100 TI - Receptor for 1,25-dihydroxyvitamin D in a vascular smooth muscle cell line derived from rat aorta. AB - The presence of a specific receptor for 1,25-dihydroxyvitamin D (1,25(OH)2D) was investigated in a cell line A7r5 derived from fetal aorta. 1,25(OH)2[3H]D3 binding to cytosol was saturated at 0.6-1 nM, and Scatchard analysis yielded dissociation constant and binding sites, (3.02 +/- 0.4) X 10(-11) M and 33.9 +/- 5.8 fmol/mg protein, respectively. Sucrose density gradient analysis revealed the sedimentation constant 3.2 S. Furthermore, the receptor protein had affinity for DNA-cellulose column and eluted with 0.2 M KCl. These data suggest that vascular smooth muscle cell may be a target tissue of vitamin D. PMID- 3038101 TI - Interaction of tetrapyrrolic rings with rhodamine 110 and 123 and with rhodamine 110 derivatives bearing a peptidic side chain. AB - Rhodamines 110 and 123, and rhodamine 110 linked via peptide bonds to Arg, Cbz Arg and Cbz-Ile-Pr-Arg interact with free base porphyrins or cytochrome C. Rhodamines 110 and 123 essentially form 1:1 complexes while the other derivatives form 2:1 complexes. The possible biological implications of these results are discussed. PMID- 3038102 TI - Inhibition studies of three classes of Desulfovibrio hydrogenase: application to the further characterization of the multiple hydrogenases found in Desulfovibrio vulgaris Hildenborough. AB - The three types of hydrogenase hitherto characterized in genus Desulfovibrio exhibit distinctive inhibition patterns of their proton-deuterium exchange activity by CO, NO and NO2-. The (Fe) and (NiFeSe) hydrogenases are the most sensitive to all three inhibitors while the (NiFe) enzymes, relatively little inhibited by CO, are still very sensitive to NO but unaffected by NO2-. These differences together with some specific catalytic properties, in particular the pH profile and the H2 to HD ratio in the exchange reaction, constitute a simple means of characterizing multiple hydrogenases present in one or different species. PMID- 3038103 TI - Red cell membrane (Na+ +K+)-ATPase in diabetes mellitus. AB - The red cell membrane (Na+ +K+)-ATPase activity is significantly elevated in diabetes mellitus. The osmotic fragility of diabetic red cells is also increased. In vivo insulin treatment restores the enzyme activity and the osmotic fragility to the normal level. In vitro insulin treatment of diabetic red cells was found to inhibit the further increase in its activity, but it failed to restore the activity to the normal level as in vivo. In diabetes increased Km of (Na+ +K+) ATPase for ATP was observed but Vmax remained the same. Arrhenius plot of this enzyme was also altered in diabetes. PMID- 3038104 TI - Erythroid differentiation factor stimulates hydrolysis of polyphosphoinositide in Friend erythroleukemia cells. AB - We examined an early action of erythroid differentiation factor (EDF), a polypeptide which induces differentiation of Friend murine erythroleukemia (MEL) cells. (Eto et al., Biochem. Biophys. Res. Commun. 142: 1095-1103, 1987). In MEL cells, EDF caused a rapid and transient increase in cytoplasmic concentration of free calcium, [Ca2+]c. EDF increased [Ca2+]c even in the absence of extracellular calcium. When [3H]inositol-labeled MEL cells were incubated with EDF, EDF rapidly increased radioactivity in inositol trisphosphate, bisphosphate and monophosphate. EDF also increased [3H] diacylglycerol in [3H]arachidonate-labeled MEL cells. These results indicate that EDF increases [Ca2+]c by stimulating hydrolysis of polyphosphoinositide. PMID- 3038106 TI - A point mutation at codon 13 of the N-ras oncogene in a human stomach cancer. AB - A surgically removed human stomach cancer with the histological diagnosis of poorly differentiated adenocarcinoma contained an activated N-ras oncogene detected by an in vivo selection assay in nude mice using transfected NIH3T3 cells. Analysis using synthetic 20-mer oligonucleotide probes revealed a point mutation from G to C at the first letter of codon 13 of the N-ras gene resulting in the substitution of arginine for glycine. This is the first observation of an activated N-ras oncogene in human stomach cancers. PMID- 3038105 TI - Comparative effects of polymyxin B, phorbol ester and bryostatin on protein phosphorylation, protein kinase C translocation, phospholipid metabolism and differentiation of HL60 cells. AB - The effects of protein kinase C (PKC) inhibitor polymyxin B (PMB) and PKC activators 12-O-tetradecanoylphorbol-13-acetate (TPA) and bryostatin on intact HL60 cells were examined. It was found that each of the three agents exhibited similar effects on phosphorylation of certain endogenous proteins, PKC translocation from cytoplasm to plasma membrane and formation of CDP-choline. TPA, however, was the only agent that stimulated phosphatidylcholine formation. Differentiation of HL60 cells was potently induced by TPA; in comparison bryostatin was a relatively weaker inducer and PMB was without effect. The data indicated that the effects of the PKC inhibitor PMB on intact cells could not be predicted by its in vitro activity, and that certain TPA-dependent but PKC independent reactions might be crucial in HL60 cell differentiation. PMID- 3038107 TI - NADP+ phosphatase: a novel mitochondrial enzyme. AB - Mitochondria contain a NADP+ phosphatase in the matrix space. This is shown by both incubation of mitochondria and subfractions derived thereof with added NADP+ and by analysis of endogenous pyridine nucleotides after enzymatic oxidation in Ca2+-loaded mitochondria. The apparent KM for NADP+ is about 1.2 mM. NADPH is not a substrate. The enzyme may be important for modulation of posttranslational modification of macromolecules in mitochondria. PMID- 3038108 TI - Effect of a collagen-derived octapeptide on phosphoinositide turnover and 43K protein phosphorylation in collagen-activated platelets. AB - A collagen-derived octapeptide KPGEPGPK which specifically inhibits the activation of platelets by collagen has been tested for its ability to affect the collagen-induced phosphoinositide breakdown and protein phosphorylations. Collagen produced a transient decrease followed by a rapid resynthesis of [32P] phosphatidyl 4-5 bisphosphate (PIP2) and 4-mono phosphate (PIP). Octapeptide, at a concentration preventing aggregation but allowing shape change, did not impair the phosphoinositide breakdown, whereas the P43 phosphorylation was strongly inhibited. Higher concentrations of peptide which did not permit any shape change were needed to hinder the PIP2 and PIP decrease. Therefore, the octapeptide appears to affect early events of the collagen-induced platelet activation involving the P43 phosphorylation, independently of its effect on the receptor stimulated phosphoinositide hydrolysis. PMID- 3038109 TI - Early and late mitogenic events induced by FGF on bovine epithelial lens cells are not triggered by hydrolysis of polyphosphoinositides. AB - Basic or acidic forms of FGF, a potent mitogen for Bovine Epithelial Lens cells caused a rapid and transient rise in cytoplasmic Ca2+ followed by an increase in intracellular pH of 0.4 units. When cells were labeled at equilibrium with [3H] inositol, no significant breakdown of polyphosphoinositides (in the presence of 20 mM LiCl) could be detected in response to 10-100 ng/ml of FGF. Similarly, fetal calf serum efficiently reinitiated DNA synthesis in these cells with little stimulation of polyphosphoinositide hydrolysis. In contrast, prostaglandin F2 alpha and angiotensin II, two weak mitogens for BEL cells, were found potent agonists of polyphosphoinositide breakdown. These results strongly indicate that the mitogenic action of FGF is not coupled to phospholipase C activation, a conclusion consistent with the fact that the FGF-induced [Ca2+]i rise is strictly dependent upon external Ca2+. PMID- 3038110 TI - Direct evidence for the modulation of human platelet cytosolic free Ca2+ by intracellular cyclic AMP produced with a photoactivatable derivative. AB - We have previously reported that intraplatelet "cyclic AMP jumps" produced with newly synthesized photoactivatable cyclic AMP analogue, inhibited washed rat platelet aggregation and serotonin release as induced by thrombin. Using the same approach on human platelets, thrombin-induced platelet aggregation was dose dependently inhibited only when a flash was delivered. The mechanism of action of intraplatelet cyclic AMP as resulting from photolysis could be by controlling the level of cytosolic Ca2+. In order to test this hypothesis, the same protocol was used on human platelets preloaded with the internal Ca2+ fluorescent indicator, Quin 2, we found that the extent and the rate of the rise of the cytosolic Ca2+ induced by thrombin were dramatically decreased, in the presence of the photoactivatable cyclic AMP, only following photoirradiation. In addition, the flashes were produced, in the presence of photoactivatable cyclic AMP, after the thrombin-induced rise of internal Ca2+ had reached its peak. In these conditions, photoirradiation caused a rapid fall in fluorescence. These experiments provide the first direct evidence that intracellular cyclic AMP is involved in the control of platelet cytosolic Ca2+ by inhibition of its mobilization and by stimulation of its sequestration. PMID- 3038111 TI - Cloning and sequencing of Plasmodium falciparum DNA fragments containing repetitive regions potentially coding for histidine-rich proteins: identification of two overlapping reading frames. AB - DNA sequences, potentially coding for histidine-rich proteins, were isolated from a P. falciparum genomic library using an oligonucleotide probe consisting of histidine codon repeats. Sequencing revealed that the different DNA fragments contain long repetitive regions very homologous to the probe. One clone was fully sequenced and contains two open reading frames that overlap in the repetitive region but are located on opposite strands. Analysis suggests that both are coding. One frame could code for a small histidine-rich protein, the other for a protein containing many aspartic acid residues. Southern blotting revealed that these sequences are conserved in all three P. falciparum strains studied. PMID- 3038112 TI - The expression of functional erythropoietin receptors on an interleukin-3 dependent cell line. AB - We report the expression of the erythropoietin receptors on an interleukin-3 dependent cell line. By the transfer into medium supplemented with erythropoietin, DA-1 cells were converted to an erythropoietin dependent growth state. [125I] erythropoietin was used to detect receptors specific for this hormone on the cell surface. Binding studies revealed that erythropoietin bound to 131 +/- 23 receptors/cell with a Kd of 0.54 +/- 0.2 nM. When the cells were incubated at 37 degrees C, trichloroacetic acid soluble radioactivity appeared in the medium after [125I] erythropoietin binding began to decrease, suggesting that the decline represents the degradation of cell associated [125I] erythropoietin- receptor complexes. PMID- 3038113 TI - Hepatocarcinogens induce decrease in mRNA transcripts of receptors for insulin and epidermal growth factor in the rat liver. AB - Gene expression of the receptors for insulin and epidermal growth factor (EGF) was studied in the livers of rats after a single injection of a hepatocarcinogen diethylnitrosamine (DEN) 100 mg/kg or after feeding the animals N acetylaminofluorene (AAF) 0.02% w/w. DEN induced a time-dependent decrease in mRNA transcription of the receptors for insulin (10.3 and 8.5 Kb) and EGF (10.0, 5.8 and 2.8 Kb), evident already after 4 hours, reaching a nadir of 10-20% of the initial level between 16 and 24 hours and returning to normal by 10 days. In rats fed AAF, transcription of both receptor genes decreased to less than 20% of the control values after 2 days. In the livers of rats treated with DEN, that developed hepatocellular carcinomas one year later, expression of both receptors was also very low. DNA showed no changes. The results suggest that the hepatocarcinogens or their metabolites decrease RNA transcription or destabilize the steady state RNA level of both receptors in the early phase of toxic effect and that some tumor-derived products may be involved in the same phenomenon in the later stage of tumor development. PMID- 3038114 TI - Hydrogen peroxide-induced glutathione depletion and aldehyde dehydrogenase inhibition in erythrocytes. AB - To study relationships between lipid peroxidation and aldehyde dehydrogenase (ALDH) inhibition, the Stocks and Dormandy model of H2O2-induced lipid peroxidation in erythrocytes was employed. Hydrogen peroxide treatment of erythrocytes and erythrocyte lysates caused a dose-dependent inhibition and depletion of ALDH and reduced glutathione (GSH) respectively. Complete ALDH inhibition and glutathione depletion occurred before significant lipid peroxidation was detected by HPLC analysis of malondialdehyde-thiobarbituric acid adducts. Hydroxyl radical scavengers did not antagonize the hydrogen peroxide induced enzyme inhibition. Studies with the iron chelator desferrioxamine suggested that the hydrogen peroxide-induced ALDH inhibition was mediated by iron in erythrocyte lysates but not in semi-purified (and Chelex-treated) ALDH preparations. Glutathione peroxidase reduction of H2O2 exhibited an anomalous GSH dependence which was not in agreement with the accepted reaction mechanism. Reduced glutathione also antagonized the hydrogen peroxide-induced ALDH inhibition by possible complex formation with the enzyme. A hypothetical model is presented which accounts for the observed responses to hydrogen peroxide. PMID- 3038115 TI - Effects of allopurinol on myocardial ischemic injury induced by coronary artery ligation and reperfusion. AB - The effects of allopurinol pretreatment (1 mg/ml in the drinking water for 7 days at an estimated daily dose of 75 mg/kg) on biochemical and chemical changes occurring following left circumflex coronary artery ligation (40 min) and reperfusion (60 min) were examined in pentobarbital-anesthetized rabbits. During the ischemic phase, allopurinol pretreatment provided significant preservation of cellular ATP levels and of mitochondrial ATP generation as compared with untreated animals (P less than 0.05). During the reperfusion phase, allopurinol pretreatment significantly prevented the decrease in left ventricular pressure, sodium and calcium accumulation and decreases in sarcolemmal Na+,K+-stimulated and sarcoplasmic reticulum K+,Ca2+-stimulated ATPase activities as compared with untreated animals (P less than 0.05). In contrast, the decrease in mitochondrial (azide-sensitive) ATPase during ischemia and the partial recovery during reperfusion were unaffected by allopurinol pretreatment. Our results indicate that the myocardial protective effects of allopurinol may differ mechanistically in the ischemic and reperfusion phases of injury. The fact that rabbit hearts do not contain detectable xanthine oxidase activity would seem to preclude an obligatory role of this enzyme both in the generation of myocardial ischemic/reperfusion injury and in the protective actions of allopurinol. PMID- 3038116 TI - In vitro pharmacologic activity of congener derivatives and model conjugates of propranolol and practolol. AB - Previous studies in our laboratory suggested that synthetized derivatives of isoproterenol and histamine could create agonists more potent and receptor and/or tissue selective than the parent compound. In the present study we have evaluated the hypothesis that our results with isoproterenol and histamine derivatives could be extended to include beta-adrenergic antagonists. With this purpose in mind, fourteen derivatives of propranolol and practolol were synthesized and tested in four in vitro systems. The congeners and conjugates were tested using biologic assays (blocking of cAMP accumulation) and/or radioligand binding assays in S-49 lymphoma cells and in rat adipocytes, heart and lung which contain beta 1 and/or beta 2 receptors. Our results indicate that structural modifications distant from the pharmacophore alter the pharmacologic profile of the parent compound. The relative potencies of the derivatives were dependent upon several key factors including the length of the methylene spacer chain and the nature of the substituents on the aromatic ring. The presence of a spacer group with four methylenes resulted in the most active compound in each series when tested on S 49 cells. The derivatives with a paramethyl toluidide group were more potent than the derivatives with a trifluoromethyl toluidide group. The dipeptide derivatives were more potent on adipocyte than S-49 cells, suggesting a preference for beta 1 receptors. Some of the same modifications that led to altered potency and which resulted in an increased receptor and/or tissue selectivity using the progenitors isoproterenol or histamine did extrapolate to the beta blockers. Our data suggest that alterations in receptor and/or tissue selectivity must be imparted by the carrier moiety of the drug and may be related to the biochemical microenvironment of the receptors. PMID- 3038117 TI - Altered protein phosphorylation in intact rat cortical synaptosomes after in vivo administration of fluphenazine. AB - Phenothiazines such as fluphenazine are able to inhibit calcium-stimulated protein kinases in vitro in both lysed and intact synaptosomes. In this study protein phosphorylation was assayed in intact synaptosomes isolated from the cerebral cortex of rats treated chronically (21 days, 10 mg/kg, i.p.) or acutely (1 hr, 10 mg/kg, i.p.) with fluphenazine. When intact synaptosomes from chronically treated animals were prelabeled with 32Pi, there were two effects on protein phosphorylation: an increase in the basal labeling of many phosphoproteins and a decrease in depolarization-evoked protein phosphorylation. Acute injections had even more pronounced effects, but the direction and nature of the effects were the same. No effects on K+-stimulated calcium entry or on protein phosphatase activity were detected. When lysed synaptosomes from chronically treated animals were labeled in the presence of [gamma-32P]ATP, a small decrease in calmodulin-dependent and cAMP-dependent protein phosphorylation was observed. The results suggest that two different in vivo mechanisms may underlie these effects, and these are discussed. We proposed that intact synaptosomes may be a good model in which to study the in vivo mechanisms of the action of fluphenazine since they appear to retain at least some effects of the drug after subcellular fractionation. PMID- 3038118 TI - Characterization of [3H]Ro5-4864 binding to "peripheral" benzodiazepine receptors in guinea pig alveolar type II cells. AB - The binding of [3H]Ro5-4864 to peripheral benzodiazepine receptors has been demonstrated in a highly purified preparation of alveolar type II cells. [3H]Ro5 4864 binding to guinea pig alveolar type II cells indicated one population of binding sites with high affinity (KD = 5.7 nM) and saturability (Bmax = 4582 fmol/mg membrane protein). PMID- 3038119 TI - Inhibition of phosphatidylinositol kinase in vascular smooth muscle membranes by adenosine and related compounds. AB - Adenosine and 5'-chloro-5'-deoxyadenosine inhibited the phosphorylation of phosphatidylinositol in membranes prepared from aortic smooth muscle. The nucleosides did not affect the breakdown of phosphatidylinositol-4-phosphate. Under certain conditions, the membrane-bound phosphatidylinositol kinase phosphorylated exogenous phosphatidylinositol. The nucleosides inhibited the enzyme competitively with respect to magnesium-ATP and non-competitively with respect to phosphatidylinositol. Adenosine analogs modified in the ribose moiety were inhibitors with potencies comparable to that of adenosine, whereas adenine nucleotides and purine-modified adenosine analogs were much weaker inhibitors. Density gradient fractionation studies showed that phosphatidylinositol kinase is primarily associated with the sarcoplasmic reticulum. Vascular smooth muscle contraction is associated with increased phosphatidylinositol turnover. Inhibition of phosphatidylinositol kinase by intracellular adenosine may, therefore, be a factor involved in regulating vasodilation. PMID- 3038120 TI - Interactions of beta-adrenergic receptors with a membrane protein other than the stimulatory guanine nucleotide-binding protein. AB - Beta-adrenergic receptors on membranes prepared from rat lung, wild-type S49 lymphoma cells, and the adenylate cyclase-deficient variant of S49 lymphoma cells (cyc-) bind the agonist [3H]hydroxybenzylisoproterenol ([3H]HBI) with high affinity and this binding of [3H]HBI can be inhibited by GTP. Membranes prepared from these tissues were incubated with the agonist [3H]HBI or the antagonist [125I]iodopindolol ([125I]IPIN), labeled receptors were solubilized with digitonin, and the apparent molecular sizes of the ligand-bound receptor complexes were determined by high-performance size-exclusion chromatography. Results with all three tissues demonstrated that receptors labeled with [125I]IPIN were retained by the size-exclusion columns longer than receptors labeled with [3H]HBI. Thus, the apparent molecular size of soluble beta adrenergic receptors from rat lung, wild-type S49 cells, and cyc- S49 cells was larger when receptors were occupied with an agonist rather than an antagonist. The results suggest that receptors, including those on cyc- S49 cells, interact with a membrane protein, presumably a guanine nucleotide-binding protein. Since cyc- S49 cells do not contain a functional stimulatory guanine nucleotide-binding protein, but do contain a functional inhibitory guanine nucleotide-binding protein, an interaction between beta-adrenergic receptors and the inhibitory guanine nucleotide-binding protein may be responsible for the apparent increase in the molecular size of the receptor after occupation of the receptor with an agonist. PMID- 3038121 TI - Serotonergic, adrenergic and histaminergic receptors coupled to phospholipase C in cultured cerebellar granule cells of rats. AB - Phosphatidylinositol (Ptdlns) turnover has been studied in the primary culture of granule cells dissociated from the cerebellum of postnatal rat. Addition of serotonin (5-hydroxytryptamine, 5-HT) caused an increase (300-600% of control) of [3H]inositol monophosphate (IP1) accumulation in the presence of LiCl in cells prelabeled with [3H]myo-inositol. The EC50 and saturation concentrations of 5-HT were about 0.1 and 10 microM respectively. Some nonselective 5-HT receptor agonists, MK212, 5-methoxytryptamine, tryptamine and quipazine, were capable of stimulating IP1 accumulation; a selective 5-HT1A receptor agonist, 8-OH-DPAT, was ineffective. The 5-HT-induced response was potently blocked by several 5-HT2 receptor antagonists such as ketanserin, mianserin, spiroperidol and pyzotyline. The 5-HT-induced accumulation was dependent on the culturing time of granule cells with the maximal response seen in an 8-day culture. Norepinephrine (NE) also promoted an increased IP1 accumulation (300% of the control) with an EC50 of about 1 microM, while prazosin inhibited this NE-induced response with a Ki of about 0.2 nM. The responses induced by NE and 5-HT appeared to be additive. In addition, histamine in a dose-dependent manner enhanced the accumulation by about 100%; this histamine effect was blocked by triprolidine, an H1 receptor antagonist, but not by cimetidine, an H2 receptor antagonist. These results suggest that 5-HT, NE and histamine could be part of the neurotransmitter substances present in mossy fibers or other afferent nerve endings which innervate granule cells in vivo and that Ptdlns turnover regulated by their selective receptors on granule cells may play a role in modulating the excitatory function of these neurons. PMID- 3038122 TI - Automated analysis of enzyme inactivation phenomena. Application to beta lactamases and DD-peptidases. AB - In the presence of a reporter substrate, the progressive inactivation of an enzyme was easily studied by directly transmitting absorbance readings to a microcomputer. Pseudo-first order rate constants as high as 0.3 sec-1 were rapidly and accurately measured. When utilization of the reporter substrate did not exceed 10%, the rate of the reaction (vt) could be considered as proportional to the active enzyme concentration at any time during the analysis and the decrease of vt was first order with time. This simple method was used to follow the inactivation of beta-lactamases (EC 3.5.2.6) by various physical and chemical agents. When a large proportion (30-80%) of reporter substrate was destroyed, a correction was introduced to account for the corresponding decrease of its rate of utilization. This enabled experiments to be performed with a DD-peptidase and a substrate exhibiting a low delta epsilon upon hydrolysis. For the first time, the inactivation of a penicillin-sensitive enzyme by a beta-lactam could be continuously and directly observed. Finally, the method was extended to the study of hysteresis phenomena. PMID- 3038123 TI - Role of acetaldehyde in ethanol-induced increase in the activity of phosphatidate phosphatase in rat liver. AB - The effect of ethanol on the activity of phosphatidate phosphatase was studied in rat liver using membrane-bound phosphatidate and phosphatidate emulsion as substrate. A single large dose of ethanol (5 g/kg body wt) caused an increase in the enzyme activity measured with membrane-bound phosphatidate after an approximate 2-hr lag period in both cytosolic and microsomal fraction and the increase was approximately 2.2- and 1.8-fold that in control rats at 5 hr in cytosol and microsomes, respectively. A similar time-course of the increase was obtained with phosphatidate emulsion as substrate. These ethanol-induced increases in the activity of cytosolic and microsomal phosphatidate phosphatase were blocked by the pretreatment of rats with actinomycin D. The ethanol-induced rise in the activity of cytosolic and microsomal phosphatidate phosphatase measured with membrane-bound phosphatidate was abolished when rats were injected with pyrazole prior to ethanol administration. On the other hand, pretreatment with cyanamide enhanced the increase in cytosolic activity produced by a suboptimal dose of ethanol (1 g/kg), while microsomal activity was not affected by the same treatment, suggesting that acetaldehyde may be selectively involved in the ethanol-induced increase in the activity of cytosolic phosphatidate phosphatase. This hypothesis was supported by a finding that administration of paraldehyde, a cyclic trimer of acetaldehyde, produced an increase (35%) in cytosolic activity, but not in microsomal activity. PMID- 3038124 TI - Calmodulin-stimulated plasma membrane (Ca2+ + Mg2+)-ATPase: inhibition by calcium channel entry blockers. AB - Calcium channel entry blockers representing different structural classes were studied for their effects on human erythrocyte basal and calmodulin-stimulated (Ca2+ + Mg2+)-ATPase. Effects on the activity of (Mg2+)-ATPase and (Na+ + K+) ATPase were also assessed. Of the four Ca2+ entry blockers tested, only verapamil and diltiazem specifically inhibited the calmodulin-stimulated (Ca2+ + Mg2+) ATPase activity, the basal enzyme activity being unaltered by these drugs. Other membrane-associated ATPases were not affected. Calmodulin concentration effect curves showed the inhibition by verapamil (10(-3) M) and diltiazem (10(-3) M) to be non-competitive. This concentration inhibited the calmodulin-dependent increment (5.1 nM calmodulin) of the ATPase activity by 35 and 36% respectively. Similarly, both drugs inhibited the Ca2+-activation process of calmodulin stimulated activity in a non-competitive manner, decreasing Vmax by 23 and 17% respectively. Basal (Ca2+ + Mg2+)-ATPase activity was not affected by verapamil or diltiazem at any calcium concentration. In contrast, cinnarizine non specifically inhibited all four membrane ATPases including calmodulin-stimulated (Ca2+ + Mg2+)-ATPase activity at concentrations above 3 X 10(-6) M. Nifedipine was without effect on any of the four membrane ATPases. From this we conclude that certain calcium channel entry blockers can inhibit calmodulin-regulated plasma membrane Ca2+-pump ATPase. Therefore, this identifies an additional functional low affinity receptor in the plasma membrane for some of the calcium channel entry blockers. PMID- 3038125 TI - 5-Aminosalicylate: oxidation by activated leukocytes and protection of cultured cells from oxidative damage. AB - It has been postulated that oxygen radicals may play a role in the pathogenesis of inflammatory bowel disease. If so, then a drug like 5-aminosalicylate (5-ASA), which is used to treat such diseases, might work by interacting with oxygen derived species. We found that activated mononuclear cells and activated granulocytes, as well as the products of the Fenton reaction, transformed [14C]5 ASA to a number of metabolites, among which we have characterized salicylate and gentisate. We also found that the lethal effect on cultured Chinese hamster ovary cells of adding either superoxide radical or hydrogen peroxide, components of the respiratory burst of activated white blood cells, was diminished by the addition of 100 micrograms/ml (0.65 mM) of 5-ASA. Thus, we have demonstrated that 5-ASA was oxidized by the oxidative burst of white blood cells and that 5-ASA protected cells from damage by oxygen-derived species, two findings which may offer an explanation for the role of 5-ASA in the treatment of inflammatory bowel disease. PMID- 3038126 TI - Characterization of alpha 2-adrenergic receptors of calf retina membranes by [3H] rauwolscine and [3H]-RX 781094 binding. AB - Alpha 2-adrenergic receptors were identified in calf retina membranes by binding of the radiolabelled antagonists [3H]-RX 781094 and [3H]-rauwolscine. When 10 microM phentolamine was used to determine the non-specific binding, both radioligands labelled a single class of non-cooperative sites: Bmax = 1051 +/- 252 fmol/mg protein, Kd = 5.1 +/- 1.5 nM for [3H]-RX 78104 and Bmax = 1167 +/- 449 fmol/mg protein, Kd = 21.0 +/- 4.1 nM for [3H]-rauwolscine. Competition binding experiments showed the typical pharmacological potency order of alpha 2 adrenergic receptors, i.e. phentolamine greater than yohimbine greater than prazosin. Agonist competition binding curves revealed the presence of two receptor populations, having respectively high affinity (70% of the total receptor population) and low affinity for agonists, but with the same affinity for the antagonists. The high affinity sites could be converted into low affinity sites by guanine nucleotides. The non-specific binding of [3H]-RX 781094 was the same if 0.1 mM (-)-epinephrine was used instead of phentolamine. In contrast, the non-specific binding of [3H]-rauwolscine was markedly lower with (-)-epinephrine than with phentolamine. Under this condition, the Scatchard plot of [3H] rauwolscine saturation binding was curvilinear, indicating the presence of low affinity sites for the radioligand in addition to alpha 2-adrenergic receptors. Competition binding experiments revealed that these low affinity sites were distinct from adrenergic receptors: the catecholamine agonists (-)- and (-) epinephrine, (-)-norepinephrine, (-)-isoproterenol and dopamine competed with similar Ki values (microM range) whereas clonidine did not interact. Furthermore, these sites bound reserpine and the alpha 2-adrenergic antagonists yohimbine and rauwolscine but not phentolamine. PMID- 3038127 TI - Inhibition by nonsteroidal anti-inflammatory drugs of superoxide production and granule enzyme release by polymorphonuclear leukocytes stimulated with immune complexes or formyl-methionyl-leucyl-phenylalanine. AB - The effects of nonsteroidal anti-inflammatory agents on superoxide production and granule enzyme release by human polymorphonuclear leukocytes stimulated with either formyl-methionyl-leucyl-phenylalanine (fMet-Leu-Phe] or immune complexes were investigated. Cytochrome c reduction and the release of lysozyme, beta glucuronidase, myeloperoxidase and gelatinase were measured. Auranofin, phenylbutazone, sulfasalazine and the phospholipase A2 inhibitor, 4-bromophenacyl bromide, strongly inhibited these responses in fMet-Leu-Phe stimulated cells, at concentrations below 50 microM. Indomethacin, piroxicam, mefenamic acid, primaquine and quinacrine at 50-250 microM were inhibitory. Up to 1 mM ibuprofen and chloroquine inhibited superoxide production but had little effect on degranulation. With cells stimulated by IgG aggregates (immune complexes), up to 1 mM ibuprofen, mefenamic acid and piroxicam did not inhibit either response. Indomethacin, phenylbutazone, sulfasalazine and primaquine inhibited, but considerably higher concentrations were required than with fMet-Leu-Phe. Quinacrine inhibited superoxide production equally well with both stimuli but inhibited enzyme release only with fMet-Leu-Phe. Only auranofin, 4-bromophenacyl bromide, and the weakly effective chloroquine exerted approximately the same effect with both stimuli. D-Penicillamine did not affect enzyme release with either stimulus and interfered in the superoxide assay. Gelatinase release induced by fMet-Leu-Phe was affected to the same extent, or slightly more, than release of the other granule enzymes. With immune complexes, there was only modest inhibition of gelatinase release by any of the drugs at 250-1000 microM. Our results reinforce previous observations that many anti-inflammatory drugs affect neutrophil functions, but their effects vary with stimulus. The relative insensitivity of immune complex-induced responses to most of the drugs must be taken into account when considering their mode of action. PMID- 3038128 TI - (-)[125I]pindolol binding to human peripheral lung beta-receptors. AB - Beta-adrenergic receptors in human peripheral lung were characterized by biochemical and radioligand assays employing binding of the beta-antagonist ( )[125I]pindolol to plasma membrane preparations. The specific binding of ( )[125I]pindolol reached equilibrium by 45 min with an initial rate constant of 0.0282 min-1. Binding was reversible with a kinetic dissociation rate constant of 0.0146 min-1. The calculated kinetic Kd (dissociation constant) was 430 pM which agreed very well with the Kd of 394 pM obtained by Scatchard analyses of equilibrium binding data. Computer analyses of equilibrium binding experiments revealed a similar Kd of 336 +/- 24 pM. The binding capacities calculated by computer analyses (155 +/- 7 fmol/mg protein) and Scatchard analyses (113 fmol/mg protein) were also in close agreement. By all three methods (kinetic, Scatchard, and computer analyses), the data were most compatible with a single ( )[125I]pindolol binding site. Analyses of equilibrium binding data from ten different human lungs revealed values for the Kd ranging from 79 to 360 pM (mean, 136 pM), and for the receptor concentration ranging from 58 to 196 fmol/mg protein (mean, 118 fmol/mg protein). The displacement of (-)[125I]pindolol binding by various agents exhibited stereoselectivity and the expected rank order of potency predicted for interactions with beta-receptors. Isoproterenol induced a rapid and dose-related increase in cyclic AMP that was prevented by specific beta-antagonists. Approximately 70% of the beta-receptors were found to be of the beta 2-subtype by both radioligand binding and biochemical assays. Thus, ( )[125I]pindolol appears to be an excellent ligand for characterizing human lung beta-receptors since accurate and reproducible results can be obtained with this radioligand using limited tissue sample quantities. PMID- 3038129 TI - Competitive inhibition of dehydrogenases and kinases by 9-aminoacridine and quinacrine. PMID- 3038130 TI - Metabolic activation of hydralazine by rat liver microsomes. AB - There is evidence to suggest that the oxidative metabolism of hydralazine (HP), an antihypertensive drug, may represent a toxic pathway which could account for some of the adverse effects of the drug. Experiments were done to determine whether the hepatic oxidative metabolism of HP is associated with the formation of reactive metabolites. In the presence of NADPH, HP was metabolized by rat liver microsomes to three major oxidation products, phthalazine, phthalazinone (PZ), and a dimer compound. Under similar incubation conditions, radioactivity derived from [14C]HP was covalently bound to microsomal protein. Metabolite formation and covalent binding increased following pretreatment of rats with phenobarbital. In contrast, pretreatment with 3-methylcholanthrene or with the monooxygenase inhibitor, piperonyl butoxide, slightly decreased both metabolite formation and covalent binding. Electron spin resonance (ESR) analyses indicated that nitrogen-centered radicals were formed when rat liver microsomes were incubated with HP under conditions similar to those required for covalent binding and for the production of the oxidative metabolites. In addition, reduced glutathione (GSH) caused concentration-dependent decreases in the production of phthalazine, PZ, and the dimer, in the covalent binding of HP to microsomal protein, and in the formation of nitrogen-centered radicals. The results of these investigations indicate that the oxidative metabolism of HP by rat liver microsomes is highly correlated with the formation of nitrogen-centered radicals and the production of metabolites that become covalently bound to microsomal protein. These observations support the hypothesis that the oxidation of HP generates reactive metabolites which may contribute to the toxicity of the drug. PMID- 3038131 TI - Studies on the ontogenesis of the different isoenzymes of Na+, K+-ATPase in rat brain in vivo and in vitro in relation to their regulation and cellular localisation. AB - Na+,K+ ATPase isoenzyme activities (alpha(+)-high ouabain affinity; alpha low ouabain affinity) were investigated in developing rat brain in vivo and in whole rat brain reaggregating cultures in vitro. The perinatal profile of the two isoenzyme forms in vivo revealed that, although alpha activity predominates in immature (P14.5-P16) brain, the activity alpha(+) form increases more increases more rapidly such that it is predominant at birth in both cerebellum and forebrain. No regional variation in the proportional activities of the two isoenzyme forms was seen perinatally to explain the previously reported, differential sensitivity of the cerebellar alpha isoenzyme to neonatally induced hypothyroidism. Whole rat brain reaggregating cultures seeded at P16 show a normal development of Na+,K+ ATPase isoenzyme activity if grown for 14 days in a serum supplemented medium (S+). Cultivation of whole rat brain reaggregates in serum deprived medium (S-) leads to a retarded development of alpha isoenzyme activity possibly due to the absence of T3 from the medium. Hormonally-induced changes in the development of the brain Na+, K+-ATPase isoenzymes are discussed in relation to their possible function and cellular localization. PMID- 3038132 TI - Persistent synovial lymphocyte responses to cytomegalovirus antigen in some patients with rheumatoid arthritis. AB - Synovial lymphocytes from 6 of 40 patients with rheumatoid arthritis responded to cytomegalovirus antigen stimulation. 3H-thymidine uptakes were more than 3 times greater than were those of the responses to 13 other microbial antigens. Similar results were obtained in 1 patient on 7 occasions over 17 months, and in the 5 other patients on each of 2 occasions. In 3 of the 6 patients, synovial lymphocyte responses to cytomegalovirus antigen were markedly different from simultaneous peripheral blood lymphocyte responses. PMID- 3038133 TI - Eicosapentaenoic acid inhibits the secretion of triacylglycerol and of apoprotein B and the binding of LDL in Hep G2 cells. AB - The consumption of long chain polyunsaturated fatty acids by fish oils leads to profound lowering of plasma triacylglycerol but not of plasma cholesterol. Reasons for this were investigated with the human hepatoma cell line, the Hep G2 cell. Incubations with oleic acid (18:1 n9), linoleic acid (18:2 n6) and the characteristic marine fatty acid eicosapentaenoic acid (EPA, 20:5 n3) enriched cellular triacylglycerol mass, though least with EPA. However, secretion of very low density lipoprotein (VLDL) triacylglycerol and apoprotein B (measured by formation from [3H]glycerol and [3H]leucine) was markedly inhibited by EPA. Preincubation with linoleic acid reduced VLDL triacylglycerol but not apo B secretion in comparison with oleic acid which stimulated both. A possible effect on low density lipoprotein (LDL) removal was studied by measuring [125I]LDL binding. Preincubation with either EPA or linoleic acid inhibited the saturable binding of LDL, observed with oleic acid and control incubations. The binding of lipoproteins containing chylomicron remnants was not affected by any of the fatty acids. PMID- 3038134 TI - A possible antiatherogenic role for phosphocitrate through modulation of low density lipoprotein uptake and degradation in aortic smooth muscle cells. AB - The present study reports the influence of a phosphorylated polycarboxylic acid, phosphocitrate, on low density lipoprotein (LDL) metabolism in cultured rabbit aortic smooth muscle cells. Phosphocitrate profoundly influenced both LDL binding and degradation. At the maximal effective concentration (2 mM), phosphocitrate released approximately 90% of the receptor-bound [125I]LDL whilst the total amount of [125I]LDL degraded was reduced by 60%. Measurement of total cholesterol accumulation revealed that even in the presence of high concentrations of added LDL, phosphocitrate (2 mM) diminished cholesterol levels close to the basal levels seen in incubations in lipoprotein-deficient serum. Further, this inhibitory effect of phosphocitrate was demonstrable after 24 h at 37 degrees C. Phosphocitrate, a recognized anticalcifying agent, possesses a strong negative charge to size ratio at physiological pH. It is postulated that the observed effects probably arise from charge interference and/or its ability to modulate cellular calcium concentration. PMID- 3038135 TI - Increased uptake of monocyte-treated low density lipoproteins by aortic endothelium in vivo. AB - A new technique was elaborated for measuring LDL uptake by rat aortic endothelial cells in vivo, using a fluorescent marker (Dil)-labelled LDL and quantifying the fluorescence in cells selectively removed from the aorta. This technique was used to study the endothelial uptake of LDL modified by activated human monocytes (LDL A) in comparison with native LDL (LDL-N) protected from oxidation by vitamin E during the preparation. Incubation of LDL with activated monocytes increased endothelial uptake in vivo by 9.3-fold and also by 4.4-fold in cultured confluent porcine endothelium. In contrast, only a 1.5-fold increase in uptake of LDL-A was observed in sparse cultures. Cytotoxicity of monocyte-altered or native LDL did not differ as measured by the [3H]deoxyglucose-release test on cultured endothelium. Our results suggest that modification of LDL in the circulation by monocytes may make an important contribution to atherogenesis. PMID- 3038136 TI - Effects of opiate manipulations on latent inhibition in rabbits: sensitivity of the medial septal region to intracranial treatments. AB - The effects of opiate manipulations were examined on latent inhibition of classically conditioned heart rate in rabbits. Animals were exposed to an auditory stimulus that was later used as a conditioned stimulus (CS). Administration of the agonist levorphanol into the medial septal region either preceding or immediately following CS preexposure sessions attenuated latent inhibition of the conditioned response. This effect was observed in animals that showed no change in long-term retention of habituation. Administration of levorphanol into the central nucleus of the amygdala, a sensitive site for opiate manipulations on aversive conditioning tasks, was ineffective. Further analysis of the effect obtained within the septal region revealed pharmacological specificity. The effect of posttraining levorphanol was also time dependent and exhibited neuroanatomical specificity. Posttraining opiate antagonist administration into the medial septum produced a modest augmentation of latent inhibition. The medial septal region appears to provide a site where opiate manipulations alter some aspect of retention for events that occur in the absence of reinforcement. PMID- 3038137 TI - The effects of lesions to noradrenergic projections from the locus coeruleus and lateral tegmental cell groups on conditioned taste aversion in the rat. AB - Lesions of the coeruleo-cortical noradrenergic projections caused marked cortical noradrenaline depletions but were not associated with deficits in the acquisition or extinction of a conditioned taste aversion (CTA). Lesions of lateral tegmental noradrenergic projections resulted in marked hypothalamic noradrenaline depletions, enhanced neophobia to the novel taste of saccharin, unimpaired acquisition but prolonged extinction of the CTA. However, when animals with lateral tegmental noradrenergic lesions received extensive preconditioning exposure to saccharin, acquisition of the CTA was attenuated and extinction was more rapid than in controls. Alterations in CTA learning and extinction following lesions of the lateral tegmental noradrenergic system appear to reflect alterations in the way that animals with lesions react toward the hedonic aspects of taste-related stimuli rather than alterations in associational or attentional mechanisms. PMID- 3038138 TI - Preexposure to a nonaggressive opponent prevents low-intensity, social-conflict analgesia in mice. AB - In a first experiment, exposure of DBA/2 mice to a small number of attack bites by a C57BL/6 mouse resulted in low-intensity analgesia as assessed by the tail flick test. The analgesia dissipated within 10 min and was insensitive to naloxone (10 mg/kg, sc) but was antagonized by the irreversible opioid antagonist beta-chlornaltrexamine (5 mg/kg, sc). In a second experiment, preexposure to a nonaggressive C57BL/6 opponent prevented low-intensity analgesia induced by a small number of attack bites 24 hr later. The preexposure effect was abolished by naloxone (10 mg/kg, sc) given before the nonaggressive confrontation. This suggests that the release of endogenous opioids during preexposure interferes with the subsequent activation of endogenous opioid-mediated pain control mechanisms. PMID- 3038139 TI - Exposure to novelty induces naltrexone-reversible analgesia in rats. AB - The exposure of rats for 2 min to an open field, to a small box, or to inhibitory avoidance training in the small box was followed by a mild analgesia measured by the tail-flick method. The analgesia was observed as soon as 10 s after the exposure and lasted between 10 and 30 min. It was not observed in animals previously made familiar with the test situation, and it was reversed by the administration of naltrexone (0.1 mg/kg). The data suggest that novelty per se is a sufficient stimulus to activate an opioid-mediated analgesic stimulus. PMID- 3038140 TI - Dorsomedial diencephalic involvement in the juvenile play of rats. AB - Lesions of either the dorsomedial thalamus (DMT) or the parafascicular region of the thalamus (PFA) reduced rough-and-tumble social play in juvenile rats, as measured by frequency of pinning. Pinning was reduced by 33% in pups with DMT lesions and by 73% in pups with PFA lesions. Although PFA lesions had minimal effects on average pin durations, DMT pups had pins that were, on average, 105% longer than those of controls. Lesions of the PFA, but not DMT, also reduced play solicitation behaviors. Pups with PFA lesions were also insensitive to the play modulating effects of both naloxone and morphine. DMT pups were sensitive to the facilitatory effects of morphine, but naloxone was without effect in these animals. Control studies designed to evaluate general behavioral competence showed that the observed lesion effects were relatively specific to play. PMID- 3038141 TI - Opiates and medial hypothalamic knife cuts cause hyperphagia through different mechanisms. AB - Several experiments were performed to determine whether the hyperphagia caused by medial hypothalamic knife cuts and that induced by opiate agonists are mediated by a common mechanism. In the first set of experiments, male Sprague-Dawley rats were given bilateral parasagittal medial hypothalamic knife cuts or a sham procedure and fed a high-fat Crisco-chow diet. Knife-cut and sham-operated rats were approximately equally sensitive to the suppressive effects of naloxone on food intake. The kappa opiate receptor agonist ketocyclazocine generally increased daytime food intake in sham-operated rats; in contrast, the normal hyperphagia of knife-cut rats was in most cases either unchanged or decreased by ketocyclazocine. In a second set of experiments, neither diet composition nor hypothalamic knife cuts significantly affected the feeding responses to naloxone or the stimulatory effects of the kappa agonist butorphanol tartrate. It was hypothesized that the differential effects of ketocyclazocine in knife-cut and sham-operated rats are a consequence of the sedative effects of the drug combined with the elevated baseline of the knife-cut subjects. This hypothesis was supported by a third experiment, in which ketocyclazocine also reduced nocturnal intake in unoperated rats and butorphanol increased intake. That feeding responses to naloxone and butorphanol were essentially unchanged by hypothalamic knife cuts suggests that the opioid feeding system is independent of the longitudinal feeding inhibitory pathway believed to be involved in knife-cut induced hyperphagia. PMID- 3038142 TI - [Effect of a physical load on the adrenergic receptor function of peripheral blood lymphocytes and thrombocytes in healthy donors]. AB - Beta 2-adrenergic receptor density and platelet adenylate cyclase activity were determined in the peripheral blood of healthy donors during short-term physical exercise and compared to changes in biochemical parameters. The data obtained indicate the activation of sympathetic-adrenal system in general and suggest phasic regulation of catecholamine beta-adrenergic receptors. PMID- 3038143 TI - Detection of natural autoantibodies in the serum of anti-HIV-positive individuals. AB - Twenty-six sera containing anti-HIV antibodies from individuals with AIDS, AIDS related complex, lymphadenopathy syndrome, or from seropositive individuals without clinical symptoms, were tested for the presence of natural autoantibodies (NAb) to actin, DNA, tubulin, thyroglobulin, albumin, myosin and TNP-BSA. Sera from 22 seronegative individuals at high risk for HIV infection were also examined. NAb titres to all these antigens were found to be elevated in both groups as compared to the titres in pooled normal human sera. The most prevalent NAb were those of the IgG class directed against TNP found in high titre in seropositive individuals. On the basis of the above results, sera from 50 anti HIV-positive and 67 anti-HIV-negative individuals were tested for IgG antibody activity against a panel of protein antigens with different degrees of TNP substitution. The greater the TNP substitution, the higher the titres of IgG antibodies detected in anti-HIV-positive sera. Antibody titres in seronegative individuals at high risk for HIV infection were independent of the degree of TNP substitution. In almost all cases, the highest titres of IgG anti-TNP antibodies were found in anti-HIV-positive sera from individuals with clinical manifestations. PMID- 3038145 TI - Pathologic quiz case 2. Dermal cylindroma. PMID- 3038144 TI - T-cell phenotyping in the diagnosis and management of AIDS and AIDS-related disease. PMID- 3038146 TI - The natural history, pathogenesis, and treatment of juvenile angiofibroma. Personal experience with 44 patients. AB - The lessons learned from the personal care of 44 male patients suffering from juvenile angiofibroma are discussed with reference to natural history, pathogenesis, and principles of management. Many of the doctrines often accepted as based on factual evidence are discussed in relation to actual experience. In this manner, a rational view of this interesting but unusual condition is presented in the hope that some of the more bizarre opinions may now be abandoned to the benefit of future patients. PMID- 3038147 TI - Distinction between substrate- and enzyme-directed effects of modifiers of rabbit liver phosphorylase a phosphatases. AB - The natural substrate (phosphorylase a) and two alternative ones (phosphorylated histone and a tetradecapeptide consisting of residues 5-18 of rabbit skeletal muscle phosphorylase a) were used to distinguish the modes of action of some physiologically important effectors of four different molecular forms of rabbit liver phosphorlase a phosphatases. In general, glucose, caffeine, AMP, ADP, Pi, and glucose-1-P showed substrate-directed effects for the holophosphatase forms, since they usually did not affect the activity on histone phosphate and, with one slight exception (Pi), never affected the activity on the tetradecapeptide phosphate. ADP, Pi, and glucose-1-P did affect directly the relative mass (Mr) 35,000 phosphatase, in addition to an inhibition mediated via phosphorylase a. ATP exerted both substrate- and enzyme-directed effects for the Mr 35,000 phosphatase and phosphatases 1 and 2A2, but only a substrate-directed effect for phosphatase 2A1, suggesting that the gamma-subunit of the type 2 phosphatases may prevent ATP binding to the phosphatase. Mg2+ showed substrate-directed effects for phosphatases 1, 2A1, and 2A2, and an additional enzyme-directed effect for the Mr 35,000 phosphatase form. Furthermore, Mg2+ could not abolish ATP inhibition of the tetradecapeptide phosphatase activity, but significantly overcame ATP inhibition of the phosphorylase a phosphatase activity, thus suggesting that its ability to reverse the ATP effect is by a substrate-directed mechanism. The substrate-directed effects seen for the different ligands on the different phosphatase forms strongly indicate the significance of this form of control in the regulation of phosphorylase a phosphatase activities and may serve to narrow the otherwise broad substrate specificities of the major phosphorylase a phosphatase activities in mammalian tissues: phosphatases 1 and 2A. PMID- 3038148 TI - The complete amino acid sequence of rat submaxillary gland tonin does contain the aspartic acid at the active site: confirmation by protein sequence analysis. AB - The revised amino acid sequence of rat submaxillary gland tonin, a serine protease, does contain the active site Asp residue. The active site of this kallikrein-related enzyme is thus made up of the same catalytic triad (Asp, Ser, and His) found in all known serine proteases. The important Asp residue has now been localized in a 16 amino acid peptide previously reported as missing in the tonin sequence. The complete amino acid sequence thus contains 235 residues corresponding to a molecular weight of 25,658, more in agreement with previously reported molecular weights. Moreover, the revised structure led (a) to the assignment of Arg, Asn, and Val residues instead of His, Asp, and Gly at positions 63, 165, and 169, respectively; (b) to the assignment of residues occupying an overlapping sequence at positions 165-171, and finally (c) to the localization of two N-glycosylation sites at positions 82 and 165. These results further document the close relationship of tonin to the ever expanding kallikrein family. PMID- 3038149 TI - Adrenocorticotropin- and opiate-like hormones from pituitaries of the sockeye salmon Oncorhynchus nerka. AB - The pituitaries of vitellogenic sockeye salmon (Oncorhynchus nerka) were extracted with a mixture of acetone, water, and hydrochloric acid. The precipitate which formed upon the addition of a copious volume of acetone to the extract, designated acid acetone powder, was subjected to salt fractionation and desalting, followed by ion-exchange chromatography on CM-cellulose. An unadsorbed fraction (S-1) and four adsorbed fractions (S-2, S-3, S-4 and S-5) were obtained. Adrenocorticotropic activity was detected in the fractions by their ability to stimulate isolated rat adrenal decapsular cells to produce corticosterone and by their immunoreactivities in an adrenocorticotropin-specific radioimmunoassay. The steroidogenic activities of all fractions, except S-4, were blocked by corticotropin inhibiting peptide. Opiate activity was detected in the fractions by their ability to inhibit the binding of either [3H]naloxone or (D-ala2, D leu5)-[3H]enkephalin to rat brain membranes. There was a discrepancy in the potencies of the five fractions in the two opiate radioreceptor assays, indicating the presence of opiate peptides with different affinities of binding to the micron- and delta-opiate receptors of the rat brain. There was a separation between adrenocorticotropic and opiate receptor binding activities, suggesting that the activities were due to separate molecular entities. PMID- 3038150 TI - The thrombin-dependent enrichment of alkenylacyl ethanolamine phosphoglyceride with [14C]eicosapentaenoic and [3H]arachidonic acids in prelabelled human platelets. AB - The thrombin-dependent enrichment of alkenylacyl ethanolamine phosphoglyceride in [14C]eicosapentaenoic acid [( 14C]EPA) was demonstrated and compared with [3H]arachidonic acid [( 3H]AA) following the simultaneous prelabelling of individual human platelet phospholipids with these two fatty acids. The alkenylacyl, diacyl, and alkylacyl classes of ethanolamine phosphoglycerides (PE) were separated by thin-layer chromatography as their acetylated derivatives after hydrolysis of the parent phospholipid with phospholipase C. The ratios of [3H]/[14C] for the increased radioactivity appearing in alkenylacyl PE following 60 and 120 s of thrombin stimulation were the same as the corresponding ratio (2.0) found in the choline phosphoglycerides (PC) from control (unstimulated) platelets. These results suggest no significant selectivity between EPA and AA in the thrombin-stimulated transfer of these fatty acids from diacyl PC to alkenylacyl PE. The present findings may possibly bear some relevance to the altered platelet reactivity and (or) decreased thromboxane A2 formation observed in human subjects following the ingestion of marine lipid containing EPA. PMID- 3038151 TI - Regenerative activity in acute hepatitis A virus. Autoradiographic study after in vitro incorporation of 3H-thymidine. AB - The in vitro incorporation of 3H-thymidine by liver tissue obtained using needle biopsy from 14 patients with acute hepatitis A virus (AHAV) in the fully developed and recovery stage of disease was studied by the autoradiographic technique. Marked incorporation in hepatocyte nuclei was observed in the biopsies of patients in the fully developed stage, while incorporation was absent in the recovery stage of the disease. Results show that hepatocyte regeneration begins early in the course of AHAV disease, during which signs of cellular suffering and necrosis are still present. PMID- 3038152 TI - Calcium-activated neutral protease and its endogenous inhibitor in tissues of dystrophic and normal mice. AB - Calcium-activated neutral protease (milli-CANP) and its endogenous inhibitor are elevated in muscle tissues, primarily the skeletal muscle and heart, of dystrophic mice (C57BL/6J dy/dy) as compared to the control strain (C57BL/10J). Tissues showing relative increase of CANP also show significant loss of enzymes such as CK, LDH in comparison to plasma, where these enzymes register a significant increase. PK is lost minimally from these tissues, probably showing a "sparing effect." Absence of any significant change in CANP activity in the liver points to a specific role of CANP in the dystrophic process. In the skeletal muscle the endogenous CANP inhibitor registers a concomitant increase with CANP without altering the enzyme/inhibitor ratio. PMID- 3038153 TI - Intracellular vitamin A--binding proteins. PMID- 3038154 TI - Dietary tannins and salivary proline-rich proteins: interactions, induction, and defense mechanisms. PMID- 3038155 TI - From dietary glucose to liver glycogen: the full circle round. PMID- 3038156 TI - Oral nafazatrom in man: effect on inhaled antigen challenge. AB - The effect of oral nafazatrom (Bay g6575, 2 X 3 g) or placebo on inhaled antigen challenge was assessed in a double-blind study. In four subjects antigen challenge resulted in an immediate fall of 93.2 +/- 3.36% in airflow at 40% of vital capacity (Vp40) and a 45.85 +/- 4.95% reduction in forced partial expiratory volume at one second (FEV1). Neither nafazatrom nor placebo had any effect on baseline lung function or that after challenge. Leukotriene B4 was generated by ex vivo stimulus of blood with ionophore A23187, and quantified by high performance liquid chromatography (h.p.l.c.)-radioimmunoassay. No inhibition of LTB4 formation occurred ex vivo following oral nafazatrom, although addition of 10(-5) M nafazatrom to blood in vitro significantly inhibited LTB4 release. Peak plasma nafazatrom levels during the study ranged from 3.3 X 10(-7) M to 1.47 X 10(-6) M which are below the concentration (10(-5) M) at which significant 5 lipoxygenase inhibition occurs in vitro. Oral nafazatrom is ineffective as a 5 lipoxygenase inhibitor in man, probably because of poor bioavailability after administration. PMID- 3038157 TI - Epidermal growth factor receptors in non-small cell lung cancer. AB - The epidermal growth factor receptor is homologous to the oncogene erb-beta and is the receptor for a class of tumour growth factors (TGF-alpha). The clinical correlations with its expression were studied in 77 non-small cell lung cancers (NSCLC). They were stained for epidermal growth factor receptor (EGFr) by means of an indirect immunoperoxidase technique using a monoclonal antibody against the receptor. Normal lung tissue and normal bronchus were stained for comparison. Cancer tissue showed significantly increased staining compared to normal lung (P less than 0.05). Staining for EGFr in 40 squamous carcinomas was significantly stronger than in 37 specimens of other types of NSCLC (P less than 0.05), and staining in stage three NSCLC was stronger than in stage 1 and 2 (P less than 0.05). These results suggest that the presence of a high intensity of staining for EGF receptor is associated with spread of human non-small cell lung cancer and this receptor may be a suitable target for therapy. PMID- 3038158 TI - DNA analysis by flow cytometry, response to endocrine treatment and prognosis in advanced carcinoma of the breast. AB - The relationship between DNA content of mammary cancer and subsequent response to endocrine therapy was studied in 136 patients with advanced disease. All were treated with tamoxifen or ovarian ablation as first-line systemic therapy after relapse and were evaluable for response according to UICC criteria. DNA characterisation by flow cytometry was used on formalin fixed paraffin-embedded samples of tumour. Tumours were grouped according to DNA index into diploid (n = 52, 38%), 'tetraploid' (n = 46, 34%) and 'other DNA-aneuploid' (n = 38, 28%). The highest proportion of oestrogen receptor positive tumours (ER + ve) was found in the 'tetraploid' tumours (38/46, 85%, Chi-square = 8.53, P less than 0.02), and response rates, (SD + PR + CR), were 26/52 (50%), 34/46 (74%), and 15/38 (39%) respectively (Chi-square = 10.88, P less than 0.005). Patients with diploid or 'tetraploid' tumours survived longer and stayed in remission longer than those with 'other DNA-aneuploid' tumours. We suggest that 'tetraploid' or 'near tetraploid' human mammary tumours may comprise a distinct group of endocrine responsive tumours within the overall group of aneuploid tumours. The conventional interpretation of DNA histograms, grouping into diploid and aneuploid, may be masking important features of some tumour groups. PMID- 3038159 TI - Characteristics of red cell pyruvate kinase (PK) and pyrimidine 5'nucleotidase (P5N) abnormalities in acute leukaemia and chronic lymphoid diseases with leukaemic expression. AB - Red cell pyruvate kinase (PK), pyrimidine 5'nucleotidase (P5N) and reduced glutathione content (GSH) were studied in 126 untreated patients with acute leukaemia (AL, 80 cases), chronic lymphocytic leukaemia (B-CLL, 38 cases) and B cell lymphoma with leukaemic expression (LSCL, eight cases). Acute leukaemias were classified into lymphoblastic (ALL) and non-lymphoblastic (ANLL), the latter have been further sub-divided into four different variants according to FAB morphological criteria (1976). A significant decrease of PK activity was observed only in the ANLL group, leading to a clear-cut difference with the ALL group where a normal value was obtained. The decrease of P5N activity was similar in all the morphological variants of ANLL and no abnormalities in the low PEP assay system or after fructose 1,6-bisphosphate (Fru 1,6-P2) activation were observed. P5N activity was found to be significantly decreased in all groups of patients except in B-CLL, where it was normal. In regards to the different morphological groups of ANLL, a striking decrease of P5N activity was observed in the M3 variant. Although red cell GSH content was significantly increased in all groups of patients, no correlation was demonstrated between the raised GSH levels and the decreased P5N activities. PMID- 3038160 TI - In vitro and in vivo effects of treatment by platelet-activating factor on N formyl-met-leu-phe-mediated responses of polymorphonuclear leucocytes. AB - Two chemoattractants, the peptide N-formyl-met-leu-phe (FMLP), and the ether phospholipid, platelet activating factor (PAF), each stimulate a variety of in vitro responses in polymorphonuclear leucocytes (PMN). Because often more than one inflammatory mediator is active during inflammation, we determined the effect on PMN of sequential stimulation with these two agents. Before FMLP stimulation, human PMN were exposed to PAF, at concentrations which gave little or no response when administered alone. PAF enhanced FMLP-elicited superoxide release in a dose dependent fashion. Likewise, release of granular lysozyme from the cells was increased in PAF treated cells. Similar treatment with other phospholipids, including the lyso derivation of PAF, failed to produced these effects. Incubation with nordihydroguaiaretic acid, an inhibitor of arachidonic acid metabolism, had little effect on the enhancement of lysozyme release by PAF. To determine if enhancing effects by PAF might occur also in vivo, we studied rabbits receiving PAF and/or FMLP intravenously. When rabbits received 0.01 micrograms PAF (a dose which does not elicit the sustained neutropenia observed with higher doses of PAF) followed by 0.05 micrograms FMLP the absolute granulocyte count (AGC) dropped at 1 min (46 +/- 11% of original value), and continued to fall (24 +/- 12% at 10 min). Controls, treated with the suspending fluid for PAF, and then 0.05 micrograms FMLP, had a similar 1 min AGC value, but at 10 min AGC returned to 65 +/- 6.1% (P less than 0.001 for comparison of 10 min values). Thus PAF pretreatment enhanced FMLP-elicited granulocytopenia in vivo. Study of in vitro human PMN aggregation revealed that, at certain relative concentrations of PAF and FMLP, aggregation was enhanced. These studies show that both in vitro and in vivo responses of FMLP-stimulated PMN may be exaggerated by pre-exposure to PAF. PMID- 3038161 TI - Heterogeneity of IgG1 monoclonal anti-Rh(D): an investigation using ADCC and macrophage binding assays. AB - Three monoclonal IgG1 anti-Rh(D), UCH D4, ARC 7D5 and UKTS FC3, produced by Epstein-Barr virus transformed cells from Rh(D)-sensitized individuals, were compared with polyclonal single donor anti-D sera and therapeutic immunoglobulin preparations in antibody dependent cellular cytotoxicity (ADCC) and macrophage binding tests. When assayed at equal anti-D concentrations monoclonal antibodies varied considerably in their ADCC and macrophage binding activities: only UKTS FC3 showed significant activity in both assays, but these were substantially lower than those of the polyclonal anti-D sera and immunoglobulins. When examined in different combinations the monoclonal antibodies showed little synergism in mediating red cell destruction by the effector cells. Factors which might contribute to the diverse ADCC and macrophage binding activities of the monoclonal anti-Ds of the same IgG subclass are discussed. PMID- 3038162 TI - [Demonstration and classification of heterogeneity of phosphoprotein phosphohydrolase in swine spleen extracts based on sodium fluoride inhibition]. AB - The heterogeneity of phosphoprotein phosphohydrolase--Fpf in cellular free spleen extracts of pigs may be determined by sodium fluoride inhibition. NaF concentration of 5 X 10(-3) mol/l divides Fpf into 3 groups and shows a wide inhibition range from 15 to 85% of the initial activity of the enzyme. Naf concentration of 5 X 10(-4)mol/l stops Fpf activity in the range from 10 to 65% and divides it into two groups. To classify the studied material into Fpf phenotypes, fluoride quotient index--WIF has been proposed which expresses the relationship between the percentage of the inhibition caused by NaF at a concentration 5 X 10(-3) mol/l and that of the inhibition caused by NaF at a concentration of 5 X 10(-4) mol/l in the reaction mixture and in the measurement conditions described in the methods. Five groups with the following numerical values of WIF have been distinguished: 1.15 less than or equal to WIF less than 1.40-44%--group I; 1.50 less than or equal to WIF less than 1.80-20%--group II; 1.90 less than or equal to WIF less than 2.10-16%--group III; 2.20 less than or equal to WIF less than 2.50-16%--group IV; and WIF = 3-4%--group V. PMID- 3038163 TI - A family of receptors coupled to guanine nucleotide regulatory proteins. PMID- 3038164 TI - Lysophosphatidylcholines containing polyunsaturated fatty acids were found as Na+, K+-ATPase inhibitors in acutely volume-expanded hog. AB - Na+, K+-ATPase inhibitors possessing inhibitory activities against the specific binding of ouabain to Na+, K+-ATPase and 86Rb uptake into hog erythrocytes have been purified from the plasma of acutely saline-infused hog. The purifications were performed by a combination of Amberlite XAD-2 adsorption chromatography and four steps of high-performance liquid chromatography with four different types of columns. Fast atom bombardment (FAB) mass and proton NMR spectrometric studies identified the purified substances as gamma-arachidoyl- [LPCA(gamma), 34%], beta arachidoyl- [LPCA(beta), 4%], gamma-linoleoyl- (LPCL, 33%), and gamma-oleoyl- (LPCO, 25%) lysophosphatidylcholine, expressed in molar ratio in the plasma. Small amounts of gamma-docosapentaenoyl-, gamma-eicosatrienoyl-, and gamma palmitoyllysophosphatidylcholine were also detected by both FAB mass and 1H NMR spectrometric studies. Only gamma-acyl-LPC's showed inhibitory activities on Na+,K+-ATPase and ouabain-binding activities. These LPC's were effective at 100 microM levels in attaining 50% inhibition of the enzyme activity. The inhibition of Na+,K+-ATPase activity due to these compounds was always more sensitive than that of both ouabain-binding and 86Rb uptake activities. The ouabain-displacing activity in plasma due to these compounds increased with time during saline infusion. The maximal plasma level was approximately 10 times higher than that in the preinfusion plasma sample. Although these results suggest the gamma-acyl LPC's with long-chain polyunsaturated fatty acids are not simple competitive inhibitors to Na+, K+ -ATPase, these compounds could be implicated in the pathogenesis of the circulation abnormality through the modulation of membrane enzyme. PMID- 3038166 TI - Charged anesthetics selectively alter plasma membrane order. AB - Although indirect evidence supporting differential lipid fluidity in the two monolayers of plasma membranes has accumulated, unambiguous demonstration of this difference has been difficult to obtain. In the present study, the fluorescent probe 1,6-diphenyl-1,3,5-hexatriene (DPH), selective quenching of fluorescence by trinitrophenyl groups, and differential polarized phase fluorescence techniques were used to directly examine the static (order) and dynamic (rotational rate) components of lipid motion in the exofacial and cytofacial leaflets of LM fibroblast plasma membranes. The limiting anisotropy (0.137), the order parameter (0.590), and the rotational relaxation time (1.20 ns) of DPH in the plasma membranes (inner plus outer leaflet) indicated rapid but restricted probe motion in the lipid environment. However, the statics and dynamics of DPH motion in the individual monolayers were significantly (p less than 0.025) different. The limiting anisotropy, order parameter, and rotational relaxation time of DPH in the cytofacial monolayer were 0.036, 0.08, and 0.16 ns, respectively, greater than calculated for the exofacial monolayer of the LM plasma membrane. At appropriate concentrations, phenobarbital and, to a lesser degree, pentobarbital preferentially reduced the limiting anisotropy of DPH calculated for the exofacial leaflet while prilocaine reduced the limiting anisotropy of DPH in the cytofacial leaflet of LM fibroblast plasma membranes. In contrast, the putative cytofacial anesthetic procaine failed to show any preference for either leaflet. Arrhenius plots of DPH fluorescence in LM plasma membranes showed a prominent characteristic break point near 30-32 degrees C. Phenobarbital, pentobarbital, and procaine did not affect this break point while prilocaine selectively abolished it. The break point was therefore assigned to the inner monolayer of the LM plasma membrane. PMID- 3038165 TI - Is agonist self-inhibition at the nicotinic acetylcholine receptor a nonspecific action? AB - Agonist concentration-response relationships at nicotinic postsynaptic receptors were established by measuring 86Rb+ efflux from acetylcholine receptor rich native Torpedo membrane vesicles under three different conditions: integrated net ion efflux (in 10 s) from untreated vesicles, integrated net efflux from vesicles in which most acetylcholine sites were irreversibly blocked with alpha bungarotoxin, and initial rates of efflux (5-100 ms) from vesicles that were partially blocked with alpha-bungarotoxin. Exposure to acetylcholine, carbamylcholine, suberyldicholine, phenyltrimethylammonium, or (-)-nicotine over 10(8)-fold concentration ranges results in bell-shaped ion flux response curves due to stimulation of acetylcholine receptor channel opening at low concentrations and inhibition of channel function at 60-2000 times higher concentrations. Concentrations of agonists that inhibit their own maximum 86Rb+ efflux by 50% (KB values) are 110, 211, 3.0, 39, and 8.9 mM, respectively, for the agonists listed above. For acetylcholine and carbamylcholine, KB values determined from both 10-s and 15-ms efflux measurements are the same, indicating that the rate of agonist-induced desensitization increases to maximum at concentrations lower than those causing self-inhibition. For all partial and full agonists studied, Hill coefficients for self-inhibition are close to 1.0. Concentrations of agonists up to 8 times KB did not change the order parameter reported by a spin-labeled fatty acid incorporated in Torpedo membranes. We conclude that agonist self-inhibition cannot be attributed to a general nonspecific membrane perturbation. Instead, these results are consistent with a saturable site of action either at the lipid-protein interface or on the acetylcholine receptor protein itself. PMID- 3038167 TI - Kinetics of reduction by free flavin semiquinones of the components of the cytochrome c-cytochrome c peroxidase complex and intracomplex electron transfer. AB - The kinetics of reduction by free flavin semiquinones of the individual components of 1:1 complexes of yeast ferric and ferryl cytochrome c peroxidase and the cytochromes c of horse, tuna, and yeast (iso-2) have been studied. Complex formation decreases the rate constant for reduction of ferric peroxidase by 44%. On the basis of a computer model of the complex structure [Poulos, T.L., & Finzel, B.C. (1984) Pept. Protein Rev. 4, 115-171], this decrease cannot be accounted for by steric effects and suggests a decrease in the dynamic motions of the peroxidase at the peroxide access channel caused by complexation. The orientations of the three cytochromes within the complex are not equivalent. This is shown by differential decreases in the rate constants for reduction by neutral flavin semiquinones upon complexation, which are in the order tuna much greater than horse greater than yeast iso-2. Further support for differences in orientation is provided by the observation that, with the negatively charged reductant FMNH., the electrostatic environments near the horse and tuna cytochrome c electron-transfer sites within their respective complexes with peroxidase are of opposite sign. For the horse and tuna cytochrome c complexes, we have also observed nonlinear concentration dependencies of the reduction rate constants with FMNH.. This is interpreted in terms of dynamic motion at the protein-protein interface. We have directly measured the physiologically significant intra-complex one electron transfer rate constants from the three ferrous cytochromes c to the peroxide-oxidized species of the peroxidase. At low ionic strength these rate constants are 920, 730, and 150 s-1 for tuna, horse, and yeast cytochromes c, respectively. These results are also consistent with the contention that the orientations of the three cytochromes within the complex with CcP are not the same. The effect on the intracomplex electron-transfer rate constant of the peroxidase amino acid side chain(s) that is (are) oxidized by the reduction of peroxide was determined to be relatively small. Thus, the rate constant for reduction by horse cytochrome c of the peroxidase species in which only the heme iron atom is oxidized was decreased by only 38%, indicating that this oxidized side-chain group is not tightly coupled to the ferryl peroxidase heme iron. Finally, it was found that, in the absence of cytochrome c, neither of the ferryl peroxidase species could be rapidly reduced by flavin semiquinones.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3038168 TI - Conductance routes for protons across membrane barriers. AB - Simple phospholipid bilayers show a high level of permeability to protons; in spite of this fact, large proton gradients existing across such bilayers may decay very slowly. In sealed systems, the free movement of protons across a membrane barrier is severely restricted by the coincident development of a proton diffusion potential. Using the fluorescent weak acid N-[5-(dimethylamino)naphth-1 ylsulfonyl]glycine strongly buffered systems movement of the small number of protons giving rise to this electrical potential is insufficient to perturb the proton concentration gradient; significant flux of protons (and hence significant collapse of the concentration gradient) can only occur if protons traverse the membrane as part of an electroneutral complex or if there is a balancing flow of appropriate counterions. In both instances, proton flux is obligatorily coupled to the translocation of species other than protons. In weakly buffered systems, the small initial uncoupled electrogenic flux of protons may significantly alter the concentration gradient. This initial rapid gradient collapse caused by uncoupled electrogenic proton movements is then superimposed upon the residual collapse attributable to tightly coupled proton flux. The initial uncoupled electrogenic proton flux shows a temperature dependence very similar to that demonstrated for water permeation across simple lipid bilayers; upon cooling, there is a sharp decrease in flux at the temperature coinciding with the main gel liquid-crystalline phase transition of the lipid. The coupled proton flux shows a markedly different temperature dependence with no dramatic change in rate at the phase transition temperature and strong similarity to the behavior previously seen with solutes known to be permeating as electrically neutral compounds.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3038169 TI - Interaction of human calpains I and II with high molecular weight and low molecular weight kininogens and their heavy chain: mechanism of interaction and the role of divalent cations. AB - Calpain I prepared from human erythrocytes was half-maximally and maximally activated at 23 and 35 microM calcium ion, and two preparations of calpain II from human liver and kidney were half-maximally activated at 340 and 220 microM calcium ion and maximally activated at 900 microM calcium ion, respectively. High molecular weight (HMW) and low molecular weight (LMW) kininogens isolated from human plasma and the heavy chain prepared from these proteins inhibited calpain I as well as calpain II. The molar ratios of calpains to HMW kininogen to give complete inhibition of calpains were 1.4 for calpain I and 2.0 for calpain II, and those of calpains to heavy chain were 0.40-0.66 for calpain I and 0.85 for calpain II. LMW kininogen did not completely inhibit the calpains even with an excess amount of kininogen. The apparent binding ratio of calpain to HMW kininogen estimated from the disc gel electrophoretic analysis, however, was found to be 2:1, whereas those of calpain to LMW kininogen and of calpain to heavy chain were found to be 1:1. Calpains and kininogens failed to form complexes in the absence of calcium ion. In the presence of calcium ion, however, they formed the complexes, which were dissociable by the addition of ethylene glycol bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid. The minimum concentrations of calcium ion required to induce complex formation between calpain I and kininogens and calpain II and kininogens were 70 and 100 microM, respectively. Some other divalent cations such as Mn2+, Sr2+, and Ba2+ were also able to induce the complex formation between calpains and kininogens.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3038170 TI - DNA-dependent adenosinetriphosphatase B from mouse FM3A cells has DNA helicase activity. AB - We have detected at least four forms of DNA-dependent ATPase in mouse FM3A cell extracts [Tawaragi, Y., Enomoto, T., Watanabe, Y., Hanaoka, F., & Yamada, M. (1984) Biochemistry 23, 529-533]. The purified fraction of one of the four forms, ATPase B, has been shown to have DNA helicase activity by using a DNA substrate which permits the detection of limited unwinding of the helix. The DNA substrate consists of single-stranded circular fd DNA and the hexadecamer complementary to the fd DNA, which bears an oligo(dT) tail at the 3' terminus. The helicase activity and DNA-dependent ATPase activity cosedimented at 5.5 S on glycerol gradient centrifugation. The helicase required a divalent cation for activity (Mg2+ congruent to Mn2+ greater than Ca2+). The optimal concentrations of these divalent cations were 5 mM. The requirement of divalent cations of the DNA helicase activity was very similar to that for the DNA-dependent ATPase activity of ATPase B. The helicase activity was absolutely dependent on the presence of a nucleoside triphosphate. ATP was the most effective cofactor among the ribo- and deoxyribonucleoside triphosphates tested, and considerable levels of helicase activity were observed with other ribo- and deoxyribonucleoside triphosphates. The efficiency of a nucleoside triphosphate to serve as cofactor for the helicase activity correlated with the capacity of the nucleotide to serve as substrate for the DNA-dependent ATPase activity. The nonhydrolyzable ATP analogues such as adenosine 5'-O-(3-thiotriphosphate) were not effective for the helicase activity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3038172 TI - Selective outside-inside translocation of aminophospholipids in human platelets. AB - Spin-labeled analogues of phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, and sphingomyelin were added to human platelet suspensions. Due to the partial water solubility of these spin-labeled lipids which possess a relatively short beta-chain (C5), they incorporate rapidly in membranes. The orientation of the spin-labels within the platelet plasma membrane was assessed by following the spontaneous reduction at 37 and 4 degrees C due to endogenous reducing agents present in the cytosol. The rate of spontaneous reduction showed unambiguously that the labels incorporated initially in the outer leaflet of the plasma membrane and that the rate of outside-inside translocation of the aminophospholipids was faster than that of the choline derivatives. For example, at 37 degrees C, the half-time for the transverse diffusion of a phosphatidylcholine analogue was found to be of the order of 40 min, while it was less than 7 min for the phosphatidylserine analogue. At low temperatures, a fraction of the labels gave rise to a strongly immobilized ESR component. This fraction, which corresponded to 20-30% of the initial spin-label concentration, was found resistant to chemical reduction from the inner side of the membrane and also to externally added reducing agents such as ascorbate. Presumably these immobilized lipids are trapped in a gel phase formed in the outer leaflet at 4 degrees C. Cell aging, which depletes the cells of ATP, resulted in the progressive inhibition of the fast transport of the aminophospholipids from the outer to inner leaflet. Treatment of the cells with iodoacetamide completely blocked the transverse diffusion of the spin-labels.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3038171 TI - Coordinate turnover of nuclear and cytoplasmic histone messenger RNA following inhibition of DNA replication in HeLa S3 cells. AB - We have examined the metabolism of human H4 histone mRNA in the nucleus and cytoplasm of HeLa S3 cells following inhibition of DNA synthesis to address the extent to which histone mRNA stability in these cellular compartments is coupled to DNA replication. The nuclear and cytoplasmic levels of histone mRNAs encoded by the pF0108A human H4 histone gene were determined by S1 nuclease analysis using a 32P-labeled probe that could distinguish pF0108A transcripts from those of other members of the H4 histone multigene family. Hydroxyurea treatment resulted within 15 min in a 75% reduction in the level of histone H4 mRNA in the nucleus, which corresponds to the 85% decrease observed for H4 histone mRNA in the cytoplasm. The kinetics of nuclear and cytoplasmic H4 mRNA turnover following hydroxyurea treatment were also similar. Northern blot analysis using a 32P labeled mitochondrial cytochrome b probe indicated that the association of cytoplasmic RNA with the nuclear fraction was less than 0.5%. Treatment of cells with a protein synthesis inhibitor resulted in a 1.3-fold increase in nuclear H4 histone mRNA levels and a 1.5-fold increase of H4 mRNA in the cytoplasm after 45 min. Together, these results indicate that nuclear and cytoplasmic H4 histone mRNAs respond similarly to metabolic perturbations that influence message stability and that mechanisms operative in the turnover of histone mRNAs in the nucleus and cytoplasm may be similar. PMID- 3038173 TI - Binding of cytochrome c to liposomes as revealed by the quenching of fluorescence from pyrene-labeled phospholipids. AB - Resonance energy transfer from pyrene-fatty acid containing phospholipid derivatives to the heme of cytochrome c (cyt c) was used to observe the binding of this protein to liposomal membranes. Liposomes were formed of egg yolk phosphatidic acid (PA) and either egg yolk phosphatidylcholine or dipalmitoylphosphatidylcholine with 1 mol % of the fluorescent lipid. Binding of cyt c to liposomes was monitored by measuring the decrease either in the fluorescence intensity or in the lifetime of pyrene emission. The requirement for the presence of the acidic phospholipid in the membrane for the binding of cyt c could be reconfirmed. Below 5 mol % of phosphatidic acid in the membrane, no significant attachment of cyt c to liquid-crystalline bilayers was evident whereas upon increasing the concentration of PA further the association of cyt c progressively increased until a saturation was reached at about 30 mol % of phosphatidic acid. Addition of NaCl caused the fluorescence intensity and lifetimes to return to values observed in the absence of cyt c, thus revealing the dissociation of the protein from the membrane. The pyrene-labeled phosphatidic acid derivatives PPHPA and PPDPA were quenched more effectively than the corresponding phosphatidylcholines, apparently due to the direct involvement of the acidic head group in binding cyt c. When dipalmitoylphosphatidylcholine (DPPC) with 5 mol % of phosphatidic acid was used, no binding of cyt c to the liposomes above the phase transition temperature of the former lipid could be demonstrated whereas below the transition temperature (Tm) binding did take place.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3038174 TI - Effect of high pH on the spectral and catalytic properties of beef heart cytochrome oxidase. AB - Incubation of cytochrome oxidase at high pH induces changes in several spectral properties. The optical Soret maximum shifts to longer wavelength, and there is an apparent loss in intensity of the 655-nm band, effects that are normally assigned either to a spin-state transition in cytochrome a3 or to a reduction of heme a. However, magnetic circular dichroism spectra show that cytochrome a3 remains high spin and that both cytochrome a and cytochrome a3 are oxidized. At the same time, there is the appearance of a low-spin signal indicative of hydroxide-imidazole coordination which we assign as arising from a structural transition at cytochrome a, rather than at cytochrome a3, as has been proposed previously. With longer incubation times, a new copper signal appears with electron paramagnetic resonance parameters markedly different from those obtained from copper centers which have undergone denaturation. Spin quantitation establishes that this new resonance does not arise from CuA and suggests that high pH breaks the magnetic coupling present at the cytochrome a3-CuB center. A significant proportion of cytochrome a3 may be converted to a low-spin thiolate during this process. PMID- 3038175 TI - Mechanism and requirements of in vitro RNA splicing of the primary transcript from the T4 bacteriophage thymidylate synthase gene. AB - The splicing of a procaryotic precursor RNA transcribed from the T4 phage thymidylate synthase (td) gene with SP6 RNA polymerase was investigated in vitro. The intron excision-cyclization reaction increased progressively to 60 degrees C. Exon ligation, though barely detectable at the lower temperatures, was greatly enhanced at 60 degrees C. Both reactions required Mg2+. The addition of guanosine to the 5' end of an intron-exon II intermediate via a 3',5'-phosphodiester bond was essential for the ligation of exon I to exon II. The added guanosine and the first intron-encoded uridine are subsequently lost as a dinucleotide from the 5' end during cyclization of the linear form of the excised intron RNA. Exon ligation is intramolecular and occurs more readily in the nascent RNA molecule (cotranscriptionally) than in the finished transcript (posttranscriptionally). These data and the identification of various structural elements (P, Q, R, S, E, E') in the td intron that are found typically in eucaryotic class I introns firmly establish the td intron as the first example of class I intron of procaryotic origin. PMID- 3038176 TI - Proton nuclear magnetic resonance spectroscopy of human transferrin N-terminal half-molecule: titration and hydrogen-deuterium exchange. AB - The binding of Ga(III) to the proteolytically derived N-terminal half-molecule of human transferrin (HTF/2N) was studied by proton nuclear magnetic resonance spectroscopy. The pH-dependent titration curves of the histidinyl C(2) proton chemical shifts were altered upon formation of the GaIIIHTF/2N(C2O4) ternary complex. Two high-pK'a histidines failed to titrate when the metal and synergistic anion formed a complex with the protein. These results implicated two histidinyl residues as direct ligands to the metal. The rates of hydrogen deuterium exchange for the C(2) protons of certain histidinyl residues were substantially decreased by metal ion binding. The two ligand histidines were protected from exchange, and a third, low-pK'a, histidinyl residue was protected. We propose that this third histidinyl residue is involved in anion binding and may serve as the base in the putative proton-relay scheme proposed for complex formation. PMID- 3038177 TI - Characterization of disulfide bonds in recombinant proteins: reduced human interleukin 2 in inclusion bodies and its oxidative refolding. AB - Cloned cDNA of human interleukin 2 (IL-2) was expressed in Escherichia coli cells in which IL-2 formed insoluble inclusion bodies. Human IL-2 has three Cys residues, namely, Cys-58, Cys-105, and Cys-125, and native IL-2 has an intramolecular disulfide bond between Cys-58 and Cys-105. Since the formation of inclusion bodies was thought to be due to disorder in the oxidation state of these Cys residues, all intramolecular disulfide bond isomers of IL-2 were prepared by denaturation of native IL-2 to characterize the state of a disulfide bond in IL-2 in the inclusion bodies. These isomers can be separated from native IL-2, reduced IL-2, and IL-2's with intermolecular disulfide bonds by means of reversed-phase high-performance liquid chromatography. Human IL-2 produced in inclusion bodies in E. coli carrying a recombinant DNA was analyzed by HPLC and was proved to be a fully reduced form with no intra- and intermolecular disulfide bonds. Refolding of reduced IL-2 in the presence of reduced and oxidized glutathione and a low concentration of guanidine hydrochloride resulted in the formation of the biologically active IL-2 quantitatively. Further purification provided a practically pure IL-2 preparation without contamination of any disulfide bond isomers. PMID- 3038178 TI - Interaction between cytochrome c and cytochrome c peroxidase: excited-state reactions of zinc- and tin-substituted derivatives. AB - The effect of cytochrome c peroxidase (CCP) and apoCCP on the fluorescence and phosphorescence of Zn and Sn cytochrome c (cyt c) and the effect of cyt c on the fluorescence and phosphorescence of Zn CCP were examined. We found the following: The fluorescence yields of Zn and Sn cyt c were quenched by about 20% by CCP, consistent with energy transfer between the two chromophores with a separation of about 1.8 nm. The phosphorescence spectrum of Zn cyt c (but not Sn cyt c) shifts by 20 nm to the blue upon complexation with either CCP or apoCCP; at the same time the phosphorescence lifetime of Zn cyt c decreases from 12 +/- 2 to 6 ms with apoCCP addition. Zn CCP phosphorescence decay increases from 8.3 to 9.1 ms upon addition of poly(L-lysine) used to mimic cyt c. It is concluded from these results that binding of the redox partner or an analogue to Zn CCP and Zn cyt c results in a conformational change. The respective phosphorescence lifetimes of Zn and Sn cyt c were 13 and 3 ms in the absence of CCP and 1.6 and 1.1 ms in the presence of CCP; this corresponds to a quenching rate due to CCP of 519 and 570 s 1, for Zn and Sn cyt c, respectively. The phosphorescence of Zn CCP is also affected by native cyt c but is dramatically less than the complementary pair; the quenching rate constant is 17 s-1.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3038179 TI - Characterization of polymers of adenosine diphosphate ribose generated in vitro and in vivo. AB - Methods have been developed and applied to determine the size and branching frequency of polymers of ADP-ribose synthesized in nucleotide-permeable cultured mouse cells and in intact cultured cells. Polymers were purified by affinity chromatography with a boronate resin and were fractionated according to size molecular sieve high-performance liquid chromatography. Fractions were enzymatically digested to nucleotides, which were separated by strong anion exchange high-performance liquid chromatography. From these data, average polymer size and branching frequency were calculated. A wide range of polymer sizes was observed. Polymers as large as 190 residues with at least five points of branching per molecule were generated in vitro. Polymers of up to 67 residues containing up to two points of branching per molecule were isolated from intact cells following treatment with the DNA alkylating agent N-methyl-N'-nitro-N nitrosoguanidine. Cells treated with hyperthermia prior to DNA damage contained polymers of an average maximum size of 244 residues containing up to six points of branching per molecule. The detection of large polymers of ADP-ribose in intact cells suggests that alterations in chromatin organization effected by poly(ADP-ribosylation) may extend beyond the covalently modified proteins and very likely involve noncovalent interactions of poly(ADP-ribose) with other components of chromatin. PMID- 3038180 TI - Molecular exchange at the lipid-rhodopsin interface: spin-label electron spin resonance studies of rhodopsin-dimyristoylphosphatidylcholine recombinants. AB - The photoreceptor protein rhodopsin has been reconstituted with a single phospholipid species, dimyristoylphosphatidylcholine, at a range of different lipid/protein ratios, and the exchange rate at the lipid-protein interface has been determined from the electron spin resonance spectra of spin-labeled phosphatidylcholine. For recombinants with lipid/protein ratios in the range 41:1 to 102:1 (mol/mol), the electron spin resonance spectra of 1-acyl-2-[14-(4,4 dimethyloxazolidine-N-oxyl)stearoyl]-sn-glycero-3- phosphocholine consist of a fluid component similar to that found in pure lipid bilayers and a motionally restricted component corresponding to lipids whose motion is reduced by interaction with the intramembranous surface of rhodopsin. The relative proportion of the motionally restricted component increases with increasing protein content in the complex. Spectral subtraction with fluid and motionally restricted components (from fluid- and gel-phase lipid, respectively), which best fit the apparent components in the complex, reveals that 22 +/- 2 lipids per 39,000-dalton protein are motionally restricted, independent of lipid/protein ratio and of temperature. Simulation of the two-component spectra with the exchanged-coupled Bloch equations gives values for both the fraction of motionally restricted component and the exchange rate between the two components. Using fixed motionally restricted and fluid component line shapes at a given temperature, it is possible to obtain a consistent description of the lipid/protein ratio dependence of the spectra at each temperature. The number of motionally restricted lipids obtained by simulation, allowing for exchange, is 23 +/- 3 per 39,000-dalton protein, again independent of temperature and of lipid/protein ratio.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3038181 TI - Functional and structural characterization of the two beta 1-adrenoceptor forms in turkey erythrocytes with molecular masses of 50 and 40 kilodaltons. AB - We have previously described a specific protease in turkey erythrocytes that converts the larger 50-kDa (P50) form of the beta 1-adrenoceptor to a smaller 40 kDa (P40) form [Jurss, R., Hekman, M., & Helmreich, E. J. M. (1985) Biochemistry 24, 3349-3354]. Further functional and structural characterization studies of the two forms are reported here. When purified P50 and P40 receptors were compared with respect to their relative capabilities to couple in lipid vesicles with pure stimulatory G-proteins (Gs-proteins) prepared from turkey erythrocytes or rabbit liver, a faster and larger activation of Gs-proteins was observed in response to l-isoproterenol and guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S) with P40 than with P50 receptor. The kon values for P40 were 0.47 min-1 in the case of liver Gs and 0.22 min-1 in the case of erythrocyte Gs, whereas the corresponding values for P50 were 0.34 min-1 and 0.12 min-1, respectively. The binding properties of P50 and P40 forms of the receptor were not different, and desensitization of turkey erythrocytes on exposure to l-isoproterenol did not activate the protease. We furthermore ascertained that only the larger form with a molecular mass of 50 kDa carries the N-linked carbohydrates, which are removed on proteolytic conversion to the 40-kDa form and have either a triantennary or a tetraantennary nonfucosylated complex-type structure containing terminal sialyl residues. PMID- 3038182 TI - Spin trapping of precursors of thymine damage in X-irradiated DNA. AB - A spin-trapping method combined with ESR spectroscopy was utilized to obtain evidence for the presence of precursor radicals leading to damage in X-irradiated DNA. Two technical improvements were introduced to the conventional spin-trapping method to make possible its application to large molecules such as DNA: prior to X irradiation, sonolysis of aqueous DNA solution by 19.5-kHz ultrasound was made to get a highly concentrated DNA solution and to lower the viscosity of the solution; after precursor radicals in X-irradiated DNA were trapped by a spin trapping reagent, the DNA was digested to oligonucleotides by DNase I to get an ESR spectrum with a well-resolved hyperfine structure. Thus, it was recognized that the ESR spectrum obtained after X irradiation of the aqueous solution containing DNA and the nitroso spin-trapping reagent 2-methyl-2-nitrosopropane consisted of at least three sets of signals in the DNA. Identification of free radicals was made by comparing the spectrum with that of thymidine, which was precisely examined by a spin-trapping method combining two kinds of spin traps (nitroso and nitrone compounds) with liquid chromatography. As a result, all the signals were identified as the spin adducts of radicals produced at the thymine base moiety of DNA. The 5-hydroxy-5,6-dihydrothymin-6-yl radical was identified as a precursor of 5,6-dihydroxy-5,6-dihydrothymine (thymine glycol), the 6 hydroxy-5,6-dihydrothymin-5-yl radical as a precursor of 6-hydroxy-5,6 dihydrothymine, and the 5-methyleneuracil radical as a precursor of 5 (hydroxymethyl) uracil. PMID- 3038183 TI - Influence of template strandedness on in vitro replication of mutagen-damaged DNA. AB - We analyzed the ability of DNA polymerases to bypass damage on single- and double stranded templates. In vitro DNA synthesis was studied on UV-irradiated and polyaromatic hydrocarbon reacted (benzo[a]pyrenediol epoxide and oxiranylpyrene) double-stranded templates by a protocol involving initiation on a uniquely nicked circular double-stranded template. The template was prepared by treating single stranded (+)M13mp2 circular strands with mutagen and then hybridizing with restricted M13 RFmp2, followed by isolation of the nicked RFII forms. The protocol permits either (+), (-), or both strands to carry lesions. We found that the rules for termination and bypass of lesions previously observed with single stranded DNA templates also hold for double-stranded templates. Termination of synthesis occurs primarily one nucleotide 3' to the lesion in the template strand. Bypass of UV-induced lesions can be followed in a series of three partial reactions in the presence of Mn2+ and dGMP, which relax the specificity of nucleotide insertion and 3'----5' exonuclease activity, respectively. There is no evidence for greater permissivity of bypass in double-as opposed to single stranded templates. As with single-stranded templates, purines and preferentially deoxyadenosine (dA) are inserted opposite lesions. Lesions in the nontemplate strand elicit neither termination nor pausing. The addition of Rec A protein resulted in a measurable increase of bypass in this system. PMID- 3038185 TI - Binding of thrombin to thrombomodulin accelerates inhibition of the enzyme by antithrombin III. Evidence for a heparin-independent mechanism. AB - The endothelial cell surface provides a receptor for thrombin-designated thrombomodulin (TM) which regulates thrombin formation and the activity of the enzyme at the vessel wall surface by serving as a potent cofactor for the activation of protein C by thrombin. Heparin-like structures of the vessel wall have been proposed as another regulatory mechanism catalyzing the inhibition of thrombin by antithrombin III. In the present study, the interaction of antithrombin III with the thrombin-TM complex and its interference with heparin and polycations were investigated by using human components and TM isolated from the microvasculature of rabbit lung. Purified TM bound thrombin and acted as a cofactor for protein C activation. The addition of heparin (0.5 unit/mL) to the reaction mixture interfered neither with the binding of thrombin to TM nor with the activation of protein C. However, the polycations protamine (1 unit/mL) as well as polybrene (0.1 mg/mL) affected the thrombin-TM interaction. This was documented by an increase in the Michaelis constant from 8.3 microM for thrombin alone to 19.5 microM for thrombin-TM with the chromogenic substrate compound S 2238 in the presence of 1 unit/mL protamine. When the inhibition of thrombin by antithrombin III was determined, the second-order rate constant k2 = 8.4 X 10(3) M-1 s-1 increased about 8-fold in the presence of TM, implying an accelerative function of TM in this reaction. Although purified TM did not bind to antithrombin III-Sepharose, suggesting the absence of heparin-like structures within the receptor molecule, protamine reversed the accelerative effect of TM in the inhibition reaction.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3038184 TI - Cloning and sequence analysis of cDNAs encoding mammalian mitochondrial malate dehydrogenase. AB - A cDNA clone, named ppmMDH-1 and covering a part of the porcine mitochondrial malate dehydrogenase (mMDH; L-malate:NAD+ oxidoreductase, EC 1.1.1.37) mRNA, was isolated from a porcine liver cDNA library with a mixture of 24 oligodeoxyribonucleotides as a probe. The sequences of the probe were deduced from the known sequence of porcine mMDH amino acid residues 288-293. ppmMDH-1 covered the coding region for porcine mMDH amino acid residues 17-314 and the 3' untranslated region. Subsequently, mouse mMDH cDNA clones were isolated from a mouse liver cDNA library with the ppmMDH-1 cDNA as a probe. One of the clones, named pmmMDH-1 and containing a cDNA insert of about 1350 base pairs, was selected for sequence analysis, and the primary structure of the mouse precursor form of mMDH (pre-mMDH) was deduced from its cDNA sequence. The sequenced coding regions for the porcine and mouse mMDH mRNAs showed about 85% homology. When the deduced amino acid sequence of the mouse pre-mMDH was compared with that of the porcine mMDH, they shared a 95% homology, and the mouse pre-mMDH yielded a leader sequence consisting of 24 amino acid residues and a mature mMDH, consisting of 314 amino acid residues. The leader sequence contained three basic amino acid residues, no acidic residues, and no hydrophobic amino acid stretch. The mouse mMDH leader sequence was compared with those of three other rodent mitochondrial matrix proteins. PMID- 3038186 TI - An antimycin-insensitive succinate-cytochrome c reductase activity in pure reconstitutively active succinate dehydrogenase. AB - Antimycin-insensitive succinate-cytochrome c reductase activity has been detected in pure, reconstitutively active succinate dehydrogenase. The enzyme catalyzes electron transfer from succinate to cytochrome c at a rate of 0.7 mumole succinate oxidized per min per mg protein, in the presence of 100 microM cytochrome c. This activity, which is about 2% of that of reconstitutive (the ability of succinate dehydrogenase to reconstitute with coenzyme ubiquinone binding proteins (QPs) to form succinate-ubiquinone reductase) or succinate phenazine methosulfate activity in the preparation, differs from antimycin insensitive succinate-cytochrome c reductase activity detected in submitochondrial particles or isolated succinate-cytochrome c reductase. The Km for cytochrome c for the former is too high to be measured. The Km for the latter is about 4.4 microM, similar to that of antimycin-sensitive succinate-cytochrome c activity in isolated succinate-cytochrome c reductase, suggesting that antimycin-insensitive succinate-cytochrome c activity of succinate-cytochrome c reductase probably results from incomplete inhibition by antimycin. Like reconstitutive activity of succinate dehydrogenase, the antimycin-insensitive succinate-cytochrome c activity of succinate dehydrogenase is sensitive to oxygen; the half-life is about 20 min at 0 degrees C at a protein concentration of 23 mg/ml. In the presence of QPs, the antimycin-insensitive succinate cytochrome c activity of succinate dehydrogenase disappears and at the same time a thenoyltrifluoroacetone-sensitive succinate-ubiquinone reductase activity appears. This suggests that antimycin-insensitive succinate-cytochrome c reductase activity of succinate dehydrogenase appears when succinate dehydrogenase is detached from the membrane or from QPs. Reconstitutively active succinate dehydrogenase oxidizes succinate using succinylated cytochrome c as electron acceptor, suggesting that a low potential intermediate (radical) may be involved. This suggestion is confirmed by the detection of an unknown radical by spin trapping techniques. When a spin trap, alpha-phenyl-N-tert-butylnitrone (PBN), is added to a succinate oxidizing system containing reconstitutively active succinate dehydrogenase, a PBN spin adduct is generated. Although this PBN spin adduct is identical to that generated by xanthine oxidase, indicating that a perhydroxy radical might be involved, the insensitivity of this antimycin insensitive succinate-cytochrome c reductase activity to superoxide dismutase and oxygen questions the nature of this observed radical. PMID- 3038187 TI - Na+ currents generated by the purified (Na+ + K+)-ATPase on planar lipid membranes. AB - Purified (Na+ + K+)-ATPase from pig kidney was attached to black lipid membranes and ATP-induced electric currents were measured as described previously by Fendler et al. ((1985) EMBO J. 4, 3079-3085). An ATP concentration jump was produced by an ultraviolet-light flash converting non-hydrolysable caged ATP to ATP. In the presence of Na+ and Mg2+ this resulted in a transient current signal. The pump current was not only ATP dependent, but also was influenced by the ATP/caged ATP ratio. It was concluded that caged ATP binds to the enzyme (and hence inhibits the signal) with a Ki of approx. 30 microM, which was confirmed by enzymatic activity studies. An ATP affinity of approx. 2 microM was determined. The addition of the protonophore 1799 and the Me+/H+ exchanger monensin made the bilayer conductive leading to a stationary pump current. The stationary current was strongly increased by the addition of K+ with a K0.5 of 700 microM. Even in the absence of K+ a stationary current could be measured, which showed two Na+ affinities: a high-affinity (K0.5 less than or equal to 1 mM) and a low-affinity (K0.5 greater than or equal to 0.2 M). In order to explain the sustained electrogenic Na+ transport during the Na+-ATPase activity, it is proposed, that Na+ can replace K+ in dephosphorylating the enzyme, but binds about 1000-times weaker than K+. The ATP requirement of the Na+-ATPase was the same (K0.5 = 2 microM) with regard to the peak currents and the stationary currents. However, for the (Na+ + K+)-ATPase the stationary currents required more ATP. The results are discussed on the basis of the Albers-Post scheme. PMID- 3038188 TI - Role of Na+/H+ antiport in intracellular pH regulation by rabbit enterocytes. AB - The steady-state intracellular pH (pHi) of isolated rabbit enterocytes was determined using 9-aminoacridine, a fluorescent weak base, and the null-point method with digitonin. When cells are incubated in a Na+-containing solution, the estimated value of pHi was in the range of 7.10-7.20, whereas it was 6.60-6.70 when cells were incubated in a Na+-free solution, indicating an important role of external Na+ in maintaining pHi at a slightly alkaline level. Pulse injection of Na+ into a Na+-free cell suspension induced a slowly developing alkalinization of pHi. The time course of the alkalinization was found to be dependent on the Na+ concentration. Li+ had the same effect as Na+, while K+ had a slight effect. Amiloride inhibited the effects of Na+ dose-dependently. These results indicate that the Na+/H+ antiport plays an important role in maintaining the pHi at a neutral or slightly alkaline level in the intact enterocytes. PMID- 3038189 TI - Differential effects of compound 48/80 on the ATPase and phosphatase activities of the Ca2+ pump of red cells. AB - The calmodulin antagonist compound 48/80 inhibits the phosphatase activity of the Ca2+-ATPase lowering its maximum velocity and leaving unaltered its apparent affinity for the substrate regardless on whether phosphatase activity is elicited by Ca2+ plus ATP or by calmodulin. Compound 48/80 has no effect on the Ki for ATP as inhibitor of the phosphatase. These results contrast sharply with the large increase that compound 48/80 induces in the apparent affinity of the regulatory site for the nucleotide of the Ca2+-ATPase and suggest that the active site for phosphatase activity is different from the regulatory site for ATP of the Ca2+ ATPase. PMID- 3038190 TI - Aminoglycoside antibiotics preferentially increase permeability in phosphoinositide-containing membranes: a study with carboxyfluorescein in liposomes. AB - The rate of release from multilamellar liposomes of the fluorescent probe carboxyfluorescein was determined as a measure of membrane permeability. Liposomes of phosphatidylcholine and different anionic phospholipids were incubated with low (1 microM) and high (3 mM) concentrations of calcium in the absence or presence of aminoglycoside antibiotics. The leakage of carboxyfluorescein into the medium was not caused by liposomal fusion as no vesicle fusion was observed in experiments with terbium and dipicolinic acid loaded liposomes. The basal rate of carboxyfluorescein release (in the absence or presence of 1 microM calcium) from all types of liposomes ranged from 0.1 to 0.3% of trapped carboxyfluorescein per hour. The presence of 3 mM calcium caused the greatest increase in the rate of carboxyfluorescein release (about 9-fold) in liposomes containing phosphatidylinositol 4,5-bisphosphate (PIP2) whereas liposomes containing the other anionic phospholipids (phosphatidylserine, phosphatidylinositol and phosphatidylinositol 4-phosphate) showed an approximate 5-fold increase. In the presence of 1 microM calcium, the aminoglycosides neomycin and gentamicin also increased the rate of carboxyfluorescein release, with PIP2-containing liposomes showing a 3-5-times greater response than the other liposomes, releasing up to 4.6% of trapped carboxyfluorescein per hour. This drug-induced release was dose-dependent and antagonized by calcium. In the presence of 3 mM calcium, 0.1 mM gentamicin or neomycin were ineffective while the drug at 1 mM affected carboxyfluorescein release from PIP2-liposomes only. The aminoglycoside antibiotics, neomycin, gentamicin, tobramycin, kanamycin, amikacin, netilmicin, as well as neamine and spectinomycin (all at 0.1 mM) showed a graded effect on the rate of carboxyfluorescein release from PIP2-containing vesicles in the presence of 0.1 mM calcium. The magnitude of the effect correlated well with the ototoxicity of the drugs previously determined directly in cochlear perfusions in the guinea pig. The study demonstrates that aminoglycoside antibiotics are capable of altering membrane permeabilities and that this effect is most pronounced if PIP2 is present in the bilayers. The excellent correlation between this membrane action and the in-situ toxicity of the drugs further establishes the specific role of PIP2 in the molecular mechanism of aminoglycoside-induced hearing loss. Moreover, it confirms the usefulness of such physicochemical models for the screening and prediction of aminoglycoside toxicity. PMID- 3038191 TI - Unfolding and flexibility in hemoproteins shown in the case of carboxymethylated cytochrome c. AB - A circular dichroism study of carboxymethylated cytochrome c has been performed to obtain further information on the structural basis responsible for the observed changes in ligand binding and redox properties of the modified cytochrome c. The results give additional evidence of local structural changes occurring in the heme environment upon rupture of the (Met-80)-heme iron bond in the modified protein. This produces no alterations of the overall molecular conformation, but results in drastic changes in redox potential. In addition, analysis of the reversible conformational transitions induced by urea in the native and the modified proteins supports the idea that the modified derivative can be considered as an 'intermediate state' between the native and the fully unfolded protein. PMID- 3038193 TI - Physiological correlation between nucleoside-diphosphate kinases and the 21-kDa guanine-nucleotide binding proteins copurified with the enzymes from the cell membrane fractions of Ehrlich ascites tumor cells. AB - The physiological correlation between nucleoside-diphosphate kinases (NDP kinases) and the 21-kDa guanine nucleotide-binding proteins (G1 and G2) which are copurified with the enzymes from the cell membrane fractions of Ehrlich ascites tumor cells has been biochemically investigated in vitro. We found that: incubation of the phosphoenzyme (enzyme-bound high-energy phosphate intermediate) of NDP-kinases (F-I and F-II) with one of the nucleoside 5'-diphosphates in the presence of 1 mM Mg2+ or 0.25 mM Ca2+ results in the rapid formation of nucleoside 5'-triphosphates without strict base specificity; GDP on the guanine nucleotide-binding proteins (G1, G2 and recombinant v-rasH p21) acts as a phosphate acceptor for the high-energy phosphates of the phosphoenzyme in the presence of 0.25 mM Ca2+; and [32P]GTP is preferentially formed from the 32P labelled phosphoenzyme F-I and GDP-bound G1 or GDP-bound recombinant v-rasH p21 protein, even if any other nucleoside 5'-diphosphates are present in the reaction mixture. Although [32P]GTP formed was bound with the guanine nucleotide-binding proteins, it was immediately hydrolyzed by the proteins themselves in the presence of 5 mM Mg2+, but not in the presence of 0.25 mM Ca2+. Available evidence suggests that NDP-kinase may be responsible for the activation of the guanine nucleotide-binding proteins (G1, G2 and p21 proteins) through phosphate transfer by the enzyme. PMID- 3038192 TI - Lipid-poor apolipoprotein A-I in Hep G2 cells: formation of lipid-rich particles by incubation with dimyristoylphosphatidylcholine. AB - Apolipoprotein A-I is a major secretory product of the human hepatoma cell line, Hep G2; approx. 70% of apolipoprotein A-I was separated from the medium as lipid poor apolipoprotein A-I in the d greater than 1.21 g/ml fraction while 30% was associated with high-density lipoproteins (HDL) of d 1.063-1.21 g/ml. The lipid poor apolipoprotein A-I contains 50% proapolipoprotein A-I which is similar to the isoform distribution in Hep G2 preformed HDL. We tested the ability of lipid poor apolipoprotein A-I from Hep G2 to form complexes with dimyristoylphosphatidylcholine (DMPC) vesicles at DMPC/apolipoprotein A-I molar ratios of 100:1 and 300:1. Lipid-poor apolipoprotein A-I was recovered in complex form while at a 300:1 ratio, 68.8 +/- 6.3% was recovered. On electron microscopy, the former complexes were small discs 16.9 nm +/- 4.5 S.D. in diameter while the latter were larger discs 21.4 +/- 4.4 nm diameter. Non-denaturing gradient gel electrophoresis of complexes formed at a 100:1 ratio had a peak in the region corresponding to 9.64 +/- 0.08 nm; these particles possessed two apolipoprotein A I molecules. At the higher ratio, 300:1, two distinct complexes were identifiable, one which banded in the 9.7 nm region and the other in the 16.9 18.7 nm region. The former particles contained two molecules of apolipoprotein A I and the latter, three molecules. This study demonstrates that lipid-poor apolipoprotein A-I which is rich in more basic isoforms forms discrete lipoprotein complexes similar to those formed by mature apolipoprotein A-I. It is further suggested that, under the appropriate conditions, precursor or nascent HDL may be assembled extracellularly. PMID- 3038194 TI - Biochemical alterations in 7,12-dimethylbenz[a]anthracene-induced mammary tumors from rats subjected to caloric restriction. AB - Caloric restriction reduces the incidence and progression of a broad spectrum of neoplastic diseases, yet little is known about the biochemical and molecular mechanisms involved. Profiles of enzyme activities of importance in cellular energy utilization were examined in 7,12-dimethylbenz[a]anthracene-induced (DMBA) mammary adenocarcinomas from rats fed ad libitum or calorically restricted diets. The diets provided equal nutrients except for fewer carbohydrate-derived calories; graded caloric restriction was 10, 20, 30 and 40%. The specific activities of hexokinase, pyruvate kinase, lactate dehydrogenase, glucose-6 phosphate dehydrogenase, malic enzyme and fructose-1,6-bisphosphatase were all elevated to varying degrees in both large palpable and small, non-palpable tumors from calorically restricted hosts compared to activities in tumors from ad libitum-fed rats. Phosphofructokinase activity was increased in palpable tumors from calorically restricted hosts but markedly reduced in non-palpable tumors. These results suggest adaptive or compensatory alterations in tumor enzyme profiles in response to the altered nutritional state of the host. PMID- 3038195 TI - Charge-dependent regulation of NADPH oxidase activity in guinea-pig polymorphonuclear leukocytes. AB - The mechanism of respiratory burst was studied by modulating membrane surfaces with lipophilic ions in guinea-pig polymorphonuclear leukocytes and their subcellular membranes. Positively charged alkylamines in concentration ranges of 0.5 to 15 microM (ED50 values) inhibited the O2- generation with phorbol 12 myristate 13-acetate, N-formylmethionylleucylphenylalanine, A23187, myristate and arachidonate in intact cells, and the inhibition was relieved by negatively charged agents. A similar molecular size of alkylalcohols had no effects. A similar charge-dependent O2- generation was also observed with fatty acids in subcellular membrane fractions prepared from unstimulated control cells, and this was insensitive to H-7 and W-7. These results suggest that triggering of NADPH oxidase activation involves a reaction(s) that is regulated by membrane charges. PMID- 3038197 TI - Observations on the quantitation of the phosphate content of peptides by fast atom bombardment mass spectrometry. AB - Equimolar mixtures of the phosphorylated and dephosphorylated forms of several peptides have been subjected to fast-atom bombardment mass spectrometry (FABMS), to investigate whether the stoichiometry of phosphorylation can be determined from the relative molecular-ion abundances of the phospho and dephospho derivatives. It is concluded that quantitation can be achieved for peptides with large positive or negative hydrophobicity/hydrophilicity indices (delta F values) where addition of a phosphate group does not alter the distribution of the peptide within the matrix significantly. For peptides with small positive or negative delta F values, phosphopeptides tend to be partially suppressed by their dephosphorylated counterparts. Suppression can be partially or totally overcome by conversion of the peptide to a hydrophobic derivative, and by the selection of an appropriate matrix. Alternatively, addition of a very strong acid, perchloric acid, can even reverse the original suppression effect. This last effect is believed to be due to the increased ionic strength in the matrix, which forces a relatively hydrophilic analyte to the matrix surface; and the ability of such a phosphorylated analyte to form a more stable gas-phase cation. PMID- 3038196 TI - Activation of coagulation factor V by calcium-dependent proteinase. AB - Factor V is a key coagulation cofactor, regulating the rate of Factor Xa catalyzed prothrombin conversion. Activation of Factor V markedly accelerates coagulation. This study describes a new class of Factor V activators, sulfhydryl proteinases. Of the enzymes studied, calcium-dependent proteinase was the most effective activator. Activation of Factor V by this enzyme was associated with cleavage of 125I-labeled Factor V to peptides distinct from those generated by previously described activators. Calcium-dependent proteinase-activated Factor Va peptides with molecular weights of 114,000 and 93,000 bound both to Factor Xa and to cultured endothelial cells. Calcium-dependent proteinase was identified in vascular endothelial cells, a tissue that also synthesizes Factor V. These findings suggest a previously unknown mechanism for cellular regulation of coagulation. PMID- 3038198 TI - Protein kinase C activators and bradykinin selectively inhibit vasopressin stimulated cAMP synthesis in MDCK cells. AB - To evaluate a possible modulation by protein kinase C of hormonal, cAMP-mediated effects on renal epithelial cells, we studied the effect of protein kinase C activators and of bradykinin on intracellular cAMP accumulation in MDCK cells. A 15-min pretreatment of cells with phorbol 12-myristate 13-acetate or 1-oleoyl-2 acetylglycerol induced a dose-dependent inhibition of vasopressin-stimulated cAMP synthesis, but not of basal or glucagon-, prostaglandin E2-, and forskolin stimulated cAMP generation. 4 alpha-Phorbol 12,13-didecanoate, inactive on protein kinase C, did not affect cAMP accumulation. Bradykinin (0.1-10 microM) also inhibited the stimulatory effect of vasopressin on cAMP synthesis in a concentration-dependent manner, but affected neither basal cAMP content, nor its stimulation by glucagon, prostaglandin E2 and forskolin. The effect of activators of protein kinase C and of bradykinin occurred while renal prostaglandin synthesis was blocked with indomethacin. The inhibitory effect of protein kinase C activators and bradykinin on cAMP generation was reversed by the protein kinase C inhibitor H7, was enhanced by monensin, one effect of which is to block the recycling of membrane receptors, and persisted when the GTP-binding protein N1 was blocked with 1 mM Mn2+. Our data suggest that: protein kinase C can modulate the tubular effects of vasopressin by inhibiting cAMP generation; this effect is not mediated by renal prostaglandins, and might result from a direct action on the vasopressin receptor, or on its coupling with Ns; the modulation by bradykinin of vasopressin effects are likely to be exerted, at least partly, through activation of protein kinase C. PMID- 3038199 TI - Effects of epinephrine, glucagon and insulin on the activity and degree of phosphorylation of fructose-1,6-bisphosphatase in cultured hepatocytes. AB - The effects of epinephrine, glucagon and insulin on the activity and degree of phosphorylation of fructose-1,6-bisphosphatase in isolated hepatocytes maintained in cell culture for 24 h were investigated. Epinephrine caused a rapid decrease in the apparent Km monitored as the activity ratio between the activity at 12.5 and 83 microM fructose-1,6-bisphosphate, reaching a maximum after 5 min. Glucagon caused a slower and less pronounced activation, and insulin caused an equally slow increase in Km. The effect of epinephrine and glucagon was completely reciprocated by insulin and the action of insulin was totally erased by the other two. Glucagon stimulated the incorporation of [32P]phosphate into fructose-1,6 bisphosphatase from about 2.5 to 4.2 mol/mol enzyme and epinephrine to 3.5 mol/mol. The effect of the two hormones acting together was cumulative. Insulin brought about a decrease in the degree of phosphorylation to 2.0 mol/mol. The effect of epinephrine was shown to be caused by the beta-receptors, since it was completely blocked by propanolol (a beta-antagonist) and remained unaffected by the presence of phentolamine (an alpha-antagonist). PMID- 3038200 TI - Purification and properties of inositol-1,4-bisphosphate 4-phosphohydrolase from rat brain. AB - Inositol-1,4-bisphosphate 4-phosphohydrolase (inositol-1,4-bisphosphatase) was highly purified from a soluble fraction of rat brain. On SDS-polyacrylamide gel electrophoresis, the purified enzyme gave a single protein band and its molecular weight was estimated to be 42000. The isoelectric point of the enzyme was 4.3. The enzyme specifically hydrolyzed the 4-phosphomonoester linkage of inositol 1,4 bisphosphate. The Km value for inositol 1,4-bisphosphate was 30 microM, and it required Mg2+ for activity. Ca2+ was a competitive inhibitor with a Ki value of 60 microM as regards the Mg2+ binding. Li+, which is known to be a strong inhibitor of inositol 1-phosphatase (EC 3.1.3.25), inhibited the enzyme activity and caused 50% inhibition at a concentration of 1 mM (IC50 = 1 mM). Li+ was an uncompetitive inhibitor of substrate binding with a Ki value of 0.6 mM. These inhibitory parameters of Li+ were quite similar to those for inositol 1 phosphatase (IC50 = 1 mM, Ki = 0.3 mM). Thus, the effect of Li+ on decreasing the free inositol level with a subsequent decrease in agonist-sensitive phosphoinositides, is caused by its inhibition of multiple enzymes involved in conversion of inositol 1,4-bisphosphate to inositol. PMID- 3038201 TI - The structure of the promoter and amino terminal region of the pH 2.5 acid phosphatase structural gene (appA) of E. coli: a negative control of transcription mediated by cyclic AMP. AB - The regulatory and promoter regions of the gene for acid phosphatase of optimum pH 2.5 (appA) of Escherichia coli has been characterized by constructing appA lacZ protein fusions. Monitoring of beta-galactosidase activity showed that the cloned fragment harbored a region that was responsible for a negative control of transcription mediated by the cAMP-cAMP receptor (CAP) complex. The nucleotide sequence of the segment was determined. A region containing a putative CAP binding site, overlapping the -10 region of the promoter was found. Deletion analysis around the promoter region, using appA-lacZ protein fusions substantiated the hypothesis for a role of the 'consensus' sequence in the negative control mediated by cyclic AMP. In addition, the coding sequence revealed the likely existence of a signal peptide similar to the one found for other exported proteins, upstream from the phosphatase protein sequence. PMID- 3038202 TI - Heat production and respiratory enzymes in normal and runt newborn piglets. AB - The heat productions of newborn runt and normal piglets were estimated over a range of ambient temperatures. Most runts increased their heat production in the cold, but when expressed as J X kg-0.67 X min-1 it was significantly lower than in normal piglets (201.3 +/- 16.0 and 144.7 +/- 16.2 J X kg-0.67 X min-1 for controls and runts, respectively, at 32 degrees C). Runts consequently had lower deep body temperatures (37.8 +/- 0.2 and 36.8 +/- 0.3 degrees C for controls and runts, respectively, at 32 degrees C). Some runts failed to increase their metabolic rate in the cold and these had the lowest deep body temperature. Activities of respiratory enzymes in heart and diaphragm muscle were similar in all animals, whereas the longissimus dorsi of runts had significantly lower enzyme activities (76.3 +/- 4.2 and 55.5 +/- 6.6 in absorbance units for controls and runts, respectively). PMID- 3038203 TI - Temperature regulation and dopamine in schizophrenia. PMID- 3038206 TI - A modified strategy for identification of 1H spin systems in proteins. PMID- 3038204 TI - Picosecond time-resolved fluorescence of ribonuclease T1. A pH and substrate analogue binding study. AB - The tryptophyl fluorescence of ribonuclease T1 decays monoexponentially at pH 5.5, tau = 4.04 ns but on increasing pH, a second short-lived component of 1.5 ns appears with a midpoint between pH 6.5 and 7.0. Both components have the same fluorescence spectrum. Acrylamide quenches both fluorescence components, and the short-lived component is quenched fivefold faster than the predominant long component. Binding of the substrate analogue 2'-guanylic acid at pH 5.5 quenches the fluorescence by 20% and introduces a second decay component, tau = 1.16 ns. Acrylamide quenches both tryptophyl decay components, with similar quenching rates. The fluorescence anisotropy decay of ribonuclease T1 was consistent with a molecule the size of ribonuclease T1 surrounded by a single layer of water at pH 7.4, even though the anisotropy decay at pH 5.5 deviated from Stokes-Einstein behavior. The fluorescence data were interpreted with a model where the tryptophyl residue exists in two conformations, remaining in a hydrophobic pocket. The acrylamide quenching is interpreted with electron transfer theory and suggests that one conformer has the nearest atom approximately 3 A from the protein surface, and the other, approximately 2 A. PMID- 3038207 TI - [Endomorphines in schizophrenia]. AB - The biological enigma of schizophrenia has led, on the basis of thin evidence, to widen the field of clinical research to a study of endomorphines in this disease. Too many different methods of measuring the levels of opiate peptides in the CSF have been used so that it is not possible to analyse the results statistically. Clinical trials of agonists and antagonists to the opiate receptors have once again emphasised the biochemical heterogeneity of schizophrenics but have not allowed any confirmation that the endorphinical system plays any part in the genesis or symptomatology of schizophrenia. The presence of sub-groups who respond positively to experimental treatment can lead to the hope, despite the uncertainties of their mode of pharmacological action, to the next advance in the routine treatment of schizophrenia. PMID- 3038205 TI - Proton hyperfine resonance assignments using the nuclear Overhauser effect for ferric forms of horse and tuna cytochrome c. AB - Proton hyperfine resonance assignments for cytochromes c from several species are currently being successfully pursued by several laboratories. These efforts focus mostly on the ferrous forms. In contrast to that work, we have pursued assignments of the proton hyperfine shifted resonances for horse and tuna ferricytochromes c. Our results indicate that assignments are nearly identical in those two proteins. Using the pre-steady state nuclear Overhauser effect, several additional assignments have been made for the tuna protein, whereas for the horse protein, the following protons have been assigned: heme 7, alpha CH2; heme 7, beta CH2; histidine 18, beta CH2 and alpha CH; and the methionine 80, beta CH2. PMID- 3038208 TI - Epidemiological survey of HIV infection in the Veneto region of Italy. AB - A seroepidemiological survey of the Veneto region was undertaken to determine the prevalence of HIV antibodies in the sera of subjects belonging to known risk groups for AIDS and related conditions. The sera were tested by ELISA and confirmed by radioimmunoassay (Western blot). Of the AIDS patients sero-positive for HIV antibodies, 55% (6/11) were from the intravenous drug abuser group and 18% (2/11) from the homosexual group. A similar trend was seen in seropositive LAS patients, 85% (88/104) belonged to the drug abuser group and 6% (6/104) to the homosexual group. In seropositive asymptomatic subjects 83.7% (426/509) came from the drug abuser group and 4.9% (25/509) from the homosexual group. Thus drug abusers appear to be the major risk group in the Veneto region. They give rise to another risk group namely children born to seropositive drug abuser women. The seropositivity for HIV antibodies was low (0.02%) in subjects not in any risk group. Antibodies to HIV can be detected in amniotic, vitreous and cerebrospinal fluids as well as in sera. PMID- 3038209 TI - Proteins, exons and molecular evolution. AB - The discovery of the eukaryotic gene structure has prompted research into the potential relationship between protein structure and function and the corresponding exon/intron patterns. The exon shuffling hypothesis put forward by Gilbert and Blake suggests the encodement of structural and functional protein elements by exons which can recombine to create novel proteins. This provides an explanation for the relatively rapid evolution of proteins from a few primordial molecules. As the number of gene and protein structures increases, evidence of exon shuffling is becoming more apparent and examples are presented both from modern multi-domain proteins and ancient proteins. Recent work into the chemical properties and catalytic functions of RNA have led to hypotheses based upon the early existence of RNA. These theories suggest that the split gene structure originated in the primordial soup as a result of random RNA synthesis. Stable regions of RNA, or exons, were utilised as primitive enzymes. In response to selective pressures for information storage, the activity was directly transferred from the RNA enzymes or ribozymes, to proteins. These short polypeptides fused together to create larger proteins with a wide range of functions. Recent research into RNA processing and exon size, discussed in this review, provides a clearer insight into the evolutionary development of the gene and protein structure. PMID- 3038210 TI - Tyrosine kinase and phosphotyrosine phosphatase activity in human promyelocytic leukemia cells and human polymorphonuclear leukocytes. AB - Although an increase in protein phosphorylation on tyrosine was first noted as a result of cell transformation or the application of growth factors to cells, recent reports have shown high levels of tyrosine kinases in nondividing tissues. For that reason, we have investigated whether normal human polymorphonuclear leukocytes (PMN) contain tyrosine kinase and phosphatase activity. Using a copolymer of glutamine: tyrosine as a substrate for the phosphotransferase reaction, we have demonstrated that PMN contain a cytosolic tyrosine kinase activity that elutes as a single peak from Sephacryl S-200 chromatography and has a molecular weight of 70 kilodaltons. Human promyelocytic leukemia cells (HL-60), contain a similar activity (as demonstrated by column chromatography), with only 25% of the activity found in PMN. This cytosolic tyrosine kinase can phosphorylate angiotensin II and a fragment of the src protein containing tyrosine 416, which suggests a similar substrate specificity to other tyrosine phosphorylating protein kinases. In addition, we have demonstrated that PMN have double the amount of phosphotyrosine phosphatase (PTPase) activity of that found in HL-60 cells. This enzyme has a Km of 0.932 mmol/L and a Vmax of 0.355 mumol inorganic phosphate released/mg protein/min, which is similar to other cellular PTPase. Activation of PMN with f-Met-Leu-Phe and phorbol esters causes a slight but statistically significant drop in PMN PTPase activity. These results suggest that terminally differentiated myeloid cells have high tyrosine kinase and phosphatase activity, which may play a role in stimulus response coupling in the mature PMN. PMID- 3038211 TI - Productive infection by B19 parvovirus of human erythroid bone marrow cells in vitro. AB - B19 parvovirus, the cause of fifth disease and transient aplastic crisis, has been successfully propagated in suspension cultures of human erythroid bone marrow cells obtained from patients with sickle cell disease and stimulated by erythropoietin. B19 inoculation in vitro resulted in a marked decline in identifiable erythroid cells over seven to nine days of incubation. Characteristic giant early erythroid cells were seen on Wright's-Giemsa stain of infected cultures. By in situ hybridization, 30% to 40% of erythroblasts were infected at 48 hours; a similar proportion of cells showed B19 capsid protein by immunofluorescence. B19 DNA was present in erythroblasts but not in the leukocyte fraction of bone marrow. B19 replication, as determined by Southern analysis, and B19 encapsidation, as determined by sensitivity of isolated cell fractions to DNase I, were restricted to the nuclei. B19 DNA was detectable in the nuclei from infected cultures beginning at 18 hours and in the supernatant at 32 hours; B19 genome copy number was estimated at about 25,000 to 30,000/infected cell at 48 hours. Recovery of virus depended on the multiplicity of infection (moi); at low moi, approximately 200x input virus was recovered from total cultures and 50x from the culture supernatants. Virus released into the supernatant was as infectious or more infectious than virus obtained from sera of infected patients. Human erythroid bone marrow culture represents a safe in vitro system for the elucidation of the cellular and molecular biology of the pathogenic B19 parvovirus. PMID- 3038212 TI - Recombinant human granulocyte colony-stimulating factor repairs the abnormalities of neutrophils in patients with myelodysplastic syndromes and chronic myelogenous leukemia. AB - We examined the in vitro effect of recombinant human granulocyte colony stimulating factor (rhG-CSF) on neutrophil anomalies in 20 patients with myelodysplastic syndromes (MDS) and eight patients with chronic myelogenous leukemia (CML). Neutrophil alkaline phosphatase (NAP) activity was determined in nine MDS patients and eight CML patients by a scoring method. NAP scores were decreased in six of the nine patients with MDS and in all of the patients with CML. In all patients with these diseases, NAP scores increased by incubating the blood with rhG-CSF. An increase in NAP scores by rhG-CSF was observed even at a concentration of 1 U/mL in patients with MDS but was observed only at higher concentrations (1,000 to 10,000 U/mL) in patients with CML. Significant increases in NAP scores occurred at 12 hours' incubation in patients with MDS, whereas the increase was more gradual in patients with CML. This time course difference was thought to be due mainly to the difference in cell populations of circulating myeloid cells between MDS patients and CML patients. Induction of NAP activity by rhG-CSF in patients with both these diseases was suppressed by the addition of inhibitors of RNA or protein synthesis. Neutrophil superoxide anion (O2-) production induced by N-formyl-methionyl-leucyl-phenylalanine (fMLP) was determined in the other 11 patients with MDS. This neutrophil function was decreased in seven of the 11 patients with MDS, normal in two patients, and increased in two patients. Preincubation with rhG-CSF caused a significant increase in fMLP-induced O2- production in nine of the 11 patients with MDS. rhG CSF enhanced this neutrophil function in a time- and dose-dependent manner, and maximal stimulation was observed at 2,000 to 4,000 U/mL of rhG-CSF and at five to ten minutes' incubation. The present results show that rhG-CSF is able to repair at least in part the neutrophil anomalies in these patients, and our data, especially for patients with MDS, suggest the clinical usefulness of rhG-CSF for this preleukemic disorder. PMID- 3038213 TI - CML patients in blast crisis have breakpoints localized to a specific region of the BCR. AB - Chronic myelogenous leukemia (CML) is associated with the Philadelphia (Ph) chromosome, which results from a reciprocal translocation between chromosomes 9 and 22. This activates the abl oncogene by moving it from chromosome 9 and combining it with sequence located on chromosome 22. The new fusion gene, with chromosome 22 sequence at its 5' end and chromosome 9-abl sequence at its 3' end, generates a new messenger RNA (mRNA) and protein that are implicated in the pathogenesis of CML. The breakpoint near the c-abl locus on chromosome 9 can occur within a large area. In contrast, the breakpoints on chromosome 22 are concentrated within a 6 kilobase (kb) region termed the breakpoint cluster region (bcr). This study was designed to determine whether chronic-phase and blast crisis patients had identifiable differences in the structure of their Ph chromosomes. Restriction mapping of the chromosome 22 translocation breakpoints performed for 26 patients showed that the breakpoints of eight of the nine patients in blast crisis were in the 3' portion of the bcr, whereas the breakpoints in the 17 patients in the chronic phase were clustered in the 5' portion of the bcr. This suggests a strong correlation between a 3' bcr breakpoint and blast crisis in CML. PMID- 3038214 TI - Simian virus 40-transformed adherent cells from human long-term marrow cultures: cloned cell lines produce cells with stromal and hematopoietic characteristics. AB - Adherent cells from long-term marrow cultures from 23 individuals were transformed with wild-type simian virus 40 (SV40). After transformation, cloned cell lines were developed that even after rigorous subcloning invariably produced both stromal cells and round cells. The stromal cells expressed cytoskeletal filaments similar to those of long-term marrow culture adherent cells and produced interstitial and basal lamina collagen types. The round cells had the electron microscopic appearance of primitive hematopoietic cells and when examined with cytochemical stains and monoclonal antibodies to hematopoietic differentiation antigens had reaction patterns suggestive of cells from several lineages. Most round cells expressed the pan-hematopoietic T-200 determinant, and lesser percentages expressed the early T cell antigens CD-1 and CD-3, HLA-DR determinants, the monocytic antigen recognized by Leu M3, and the myeloid antigens detected by monoclonal antibodies 1G10 and 12.8. In addition, when plated in semisolid medium in the presence of a source of colony-stimulating activity, up to 11% of the cells formed colonies consisting of blastlike cells that also expressed hematopoietic cell surface determinants. The data suggest that adherent cells in long-term marrow cultures contain a cell that after transformation by SV40 obligately produces cells with hematopoietic as well as stromalike features. PMID- 3038215 TI - Effects of herpes virus carrier status on peripheral T lymphocyte subsets. AB - We studied the effects of herpes virus carrier status on peripheral blood T lymphocyte subsets in 334 healthy individuals. IgG-class antibodies against cytomegalovirus (CMV), Epstein-Barr virus (EBV), herpes simplex virus (HSV), and varicella-zoster virus (VZV) were used as markers for the carrier status of those viruses. CMV carrier status was associated with significant increases in the numbers of some T cell subsets, whereas the carrier status of EBV, HSV, and VZV had no significant effects. The 159 CMV-seropositive individuals had higher numbers of HNK1+ T cells than did the 175 CMV-seronegative individuals [mean (SD), 292 (196)/microL v 164 (89)/microL, respectively], including the CD4+HNK1+ T cells [38 (48)/microL v 9 (13)/microL, respectively] and the CD8+HNK1+ T cells [166 (146)/microL v 73 (54)/microL, respectively]. Morphological and cytochemical studies showed that the expression of HNK1 by the CD4+ and CD8+ T cells was associated with the occurrence of azurophilic cytoplasmatic granules and a loss of nonspecific esterase activity. The numbers of CD4+HNK1+ and CD8+HNK1+ T cells increased proportionally to the levels of the IgG-class CMV antibody titers. We suggest that the increased numbers of CD4+HNK1+ and CD8+HNK1+ granular T cells in CMV carriers reflect the persistent interaction between CMV and the immune system of its hosts. PMID- 3038217 TI - Use of laser-UV for inactivation of virus in blood products. AB - Inactivation of virus by UV radiation was examined as a potential method for sterilization of blood products. Samples of attenuated poliovirus, platelets and plasma were uniformly irradiated with a XeCl excimer laser that delivered 40 nsec pulses of UV at 308 nm (UVB308). Intensities and exposure does were varied from 0.11 to 1.40 MW/cm2 and 0.51 to 56.0 J/cm2, respectively. In studies conducted with low intensity UVB308 (less than or equal to 0.17 MW/cm2), using exposure doses greater than or equal to 10.8 J/cm2, it was possible to inactivate poliovirus by 4 to 6 log10. Platelets irradiated with doses less than or equal to 21.5 J/cm2 exhibited minimal damage as assessed by aggregation activity and spontaneous release of serotonin. Examination of the coagulation activity of irradiated plasma indicated that exposure doses less than or equal to 21.5 J/cm2 resulted in less than 20% increase in prothrombin and partial thromboplastin times. The use of UVB308 at a higher intensity (1.4 MW/cm2) over a similar range of exposure doses did not enhance viral inactivation but did result in increased damage to platelet and plasma proteins. These results demonstrate that at 308 nm there exists a "window of efficacy" for exposure doses between 10.8 and 21.5 J/cm2 and peak intensities less than or equal to 0.17 MW/cm2 in which a hardy virus is significantly inactivated and platelets and plasma proteins are, by functional criteria, minimally affected. Increased viral inactivation cannot be accomplished with higher UV intensities and will require additional or alternate measures. PMID- 3038216 TI - Capacity of human serum to depolymerize actin filaments. AB - Human blood depolymerizes filamentous (F-)actin. The interaction of actin filaments and monomers with human serum was studied by following the kinetics and extent of the depolymerization of pyrene-labeled F-actin and by analysis of serum proteins adhering to immobilized actin monomers. In physiologic Ca2+ concentrations, the depolymerization of F-actin proceeds in two stages: a rapid phase, attributed to direct severing of filaments by plasma gelsolin, and a slow phase attributed to the binding of actin monomers to vitamin D-binding protein (DBP). Without Ca2+, only the slow phase is observed. Human serum can completely depolymerize 10 to 18 mumol/L of actin, of which approximately 5 mumol/L occurs rapidly. Depolymerization can be accounted for by the normal serum concentrations of gelsolin and DBP. Fibrin(ogen) and fibronectin, which bind actin in vitro, do not contribute to the kinetics or extent of its depolymerization. Affinity chromatography and functional assays for the presence of gelsolin-actin complexes show that addition of G-actin to serum results in preferential formation of actin DBP complexes, but that addition of F-actin to serum produces both gelsolin-actin complexes and DBP-actin complexes. The distinctive binding of actin monomers and polymers to these two serum proteins suggests a means by which their coordinated actions are maximized in vivo, from the standpoint of depolymerizing filaments and clearing monomers from the circulation. PMID- 3038218 TI - Tumor necrosis factor (TNF) alpha: control of TNF-sensitivity and molecular mechanisms of TNF-mediated growth inhibition. PMID- 3038219 TI - [Metastatic cancers of the thyroid gland. Diagnostic difficulties]. AB - Metastatic carcinoma of the thyroid is uncommon in surgical pathology and may masquerade as primary thyroid cancer. We studied 6 cases of biopsied and/or surgically resected metastatic carcinoma of the thyroid and their corresponding primary carcinoma, with emphasis on the differential diagnosis. There were 4 men and 2 women patients aged 44 to 77. The primary carcinoma was a breast infiltrating duct carcinoma (3 cases), a colorectal adenocarcinoma (2 cases) and a bronchial oat-cell carcinoma (1 case). The interval between primary carcinoma and secondary thyroid carcinoma was 2 to 9 years in 4 cases; 2 other cases showed simultaneous occurrence. Five patients died with widespread metastases 1 to 14 months following the diagnosis of secondary carcinoma of the thyroid; 1 patient was alive after 24 months. The histological differentiation of secondary from primary thyroid cancer may be difficult in the following situations: clear-cell, Hurthle-cell and signet ring cell changes; positivity of mucins stains; production of melanin; epidermoid differentiation; very rare miscellaneous tumours ("columnar cell carcinoma" and primary thymoma of the thyroid). Immunoperoxidase methods and mucin histochemistry may help. PMID- 3038220 TI - [Breast tissue: estrogen receptors. Immunohistochemical and biochemical analysis of normal and hyperplastic tissue and carcinoma in situ. Apropos of 86 cases]. AB - Estrogen receptors (ER) were investigated using immunohistochemical techniques in 54 cases and biochemical techniques in 38 cases on a series of biopsy specimens of normal, hyperplasic and malignant mammary tissue (intraductal carcinoma). Immunohistochemical data were submitted to quantitative and semi-quantitative analysis (SAMBA 200, TITN). On normal tissue, immunostaining is bright and evenly distributed in galactophores and ductulo-lobular junctions; it is unevenly distributed but consistently present in lobules and increasing after menopause. On hyperplasic tissue, immunostaining of papillary cystadenomas and epitheliosis and fibroadenosis zones was similar to normal tissue with respect to the intensity and heterogeneity. Conversely, their immunohistochemical features are close to blunt duct adenosis and atypical lobular hyperplasia of ductulo-lobular junctions. On malignant tissue, immunostaining is unpredictable, it can be evenly distributed, unevenly distributed in patches or gradients, or totally absent. Immunohistochemical techniques can only be used to assess the intensity and distribution of ER in function of cell type in normal tissue and during a cancerous process. Thus strong receptivity of ductulo-lobular junctions may reinforce the claim that these structures are the targets for estrogen co carcinogenic substances. The heterogeneity of ER at the onset of carcinoma suggests the possibility of several pathway of development. The fact that myoepithelial cell never seem to display estrogen type receptivity makes it logical to seek a special sensitivity for them. PMID- 3038221 TI - [Reference values for maximal midexpiratory flow in black females]. AB - The aim of this publication is to contribute to the establishment of reference values for the forced expiratory flow between 25 and 75% of the vital capacity (FEF25-75) among black females in West Africa. 316 of them, aged between 10 and 70 years, were submitted to this test. They were considered free from cardiopulmonary disease after a questionnaire and clinical examination. FEF25-75 in absolute value was lower than in the white Euro-americans. It increased up to 18-19 years of age, then decreased steadily thereafter. The main equations of regression for the FEF25-75 (l X s-1) were: 10-18 years, 0.177A + 1.058 and 0.157A + 0.826H + 0.005; 19-70 years, -0.028A + 4.211 and -0.025A + 2.206H + 0.512, (where A is the age in years and H the height in metres). The results are compared with the reference values published in Black Africa, Europe and the United States. PMID- 3038222 TI - Cotrel-Dubousset instrumentation technique for revision of failed lumbosacral fusion. AB - Failed lumbosacral fusion represents a major problem for the orthopaedic surgeon. The authors report on a clinical study in which 18 patients, all previously operated on and still symptomatic, were revised using Cotrel Dubousset instrumentation to achieve rigid fixation and to correct deformities at the lumbosacral junction. In short-term follow-up, there have been no failures of hardware and the patients' clinical symptoms have improved. The surgical procedure is described, the biomechanical rationale of lumbosacral instability is analyzed, and a method of restoring physiologically normal frontal and saggital curvatures is offered. PMID- 3038223 TI - Mechanical behavior of articular cartilage quantitative changes with enzymatic alteration of the proteoglycan fraction. AB - The in-vitro viscoelastic mechanical response of normal rabbit articular cartilage is strongly dependent on the quantity and integrity of the proteoglycan fraction of the tissue matrix. Experimental results demonstrate that specific functional relationships exist between shear moduli, retardation time spectra, and proteoglycan content. Quantitative enzymolysis of the proteoglycan fraction of the tissue alters the form of these relationships in a fashion consistent with the altered physiochemical make-up of the tissue. The observed changes in mechanical behavior with controlled enzymolysis are similar to those associated with the early stages of osteoarthritis, rheumatoid arthritis, joint sepsis, and synovitis in animal models. PMID- 3038224 TI - The role of Na-hylan in reducing postsurgical tendon adhesions: Part 2. AB - Na-hylan, a chemically modified sodium hyaluronate jelly, was studied mechanically and histologically as a surgical device to diminish tendon adhesions in rabbit extensor hallucis longus tendons after severe surgical trauma. Na-hylan jelly of high viscoelastic properties was found to be highly effective in decreasing tendon adhesions. Na-hylan jelly of low viscoelastic properties was not effective in diminishing adhesion formation. There was no interference with the tensile strength of tendon healing, and no evidence of acute or chronic inflammatory response to the Na-hylan jelly. PMID- 3038225 TI - A variant of diffuse idiopathic skeletal hyperostosis: a case report. AB - The case of an elderly man with a bamboo-like spine is presented. Because the patient's osteoporosis prevented proper evaluation of the sacroiliac and apophyseal joints on plain radiographs, the case originally was misdiagnosed in an outpatient clinic as ankylosing spondylitis. A further work-up showed the integrity of the apophyseal and sacroiliac joints, as well as cortical hyperostosis of the midcervical spine consistent with the diagnosis of diffuse idiopathic skeletal hyperostosis (DISH). PMID- 3038226 TI - Periosteal ganglion case report and review of the literature. AB - The first reported case of a periosteal ganglion to be documented by computerized tomography is presented. The lesion was located on the shaft of the tibia and was unrelated to ligamentous and joint structures. A review of the literature reveals that the most common site for this lesion is the upper leg, as noted in our case; for that reason, periosteal ganglion should be included in the differential diagnosis of any soft tissue lesion in this location. CT may be more useful than plain films to demonstrate periosteal ganglion when cortical erosion is absent. PMID- 3038227 TI - Arthroscopic management of pyarthrosis. An overview. AB - The author evaluates the place of arthroscopic techniques in the treatment of bacterial infection of joints. Based on a review of the pertinent literature and an analysis of clinical material consisting of 11 cases, he concludes that pyarthrosis can be managed successfully by arthroscopic debridement combined with appropriate antibiotics and early institution of a program of range-of-motion exercises. PMID- 3038228 TI - Total hip arthroplasty and polyostotic fibrous dysplasia: a case report. AB - Polyostotic fibrous dysplasia with Albright's syndrome causes progressive skeletal lesions which require orthopaedic surgical management. A case is presented of a 43-year-old woman with a history of multiple proximal femoral osteotomies with iliac bone grafting who developed a degenerative hip joint. A total hip arthroplasty was performed and a specially designed Charnley-Mueller type of prosthesis implanted. The patient is asymptomatic eight years postoperatively. PMID- 3038229 TI - Sclerosis of a rib associated with a healed fracture of a contiguous thoracic vertebra. AB - A patient who suffered a fracture of the body of the 10th dorsal vertebra as a result of an automobile accident was examined seven years later. He complained of local pain over the gibbus site. All movements in the back and the joints of the extremities were within normal limits. Radiographs disclosed a healed compressed fracture of the body of the 10th thoracic vertebra with bony bridges contiguous with the 9th and 11th thoracic vertebrae. An unusual finding was the presence of a uniform sclerosis of a single rib at the level of the fracture site. Other bones of the rib cage, as well as the lumbar and pelvic bones, were normal. The basis for the abnormal density of the affected rib may be due to abnormal stress factors (Wolff's law). PMID- 3038230 TI - The use of clonidine for the treatment of meperidine withdrawal in a multidisciplinary pain program setting. A case presentation. AB - The management of iatrogenic drug dependence in individuals with pain can be more difficult than the treatment of the pain itself. In addition to a multidisciplinary approach to the treatment of a patient with chronic pain, there is a need for a rapid, safe, and effective method of detoxification from opiate use. Clonidine HCl, a nonopiate, has been found, in this case presentation, to be a valuable option. PMID- 3038231 TI - Monostotic fibrous dysplasia of the occipital bone: a case report. AB - Monostotic fibrous dysplasia of the occipital bone is an unusual finding. A case is presented and the differential diagnosis is discussed. PMID- 3038232 TI - Salvaging the badly placed screw. A case report. AB - On rare occasions, a surgeon who is attempting to fix a fractured hip with a sliding screw device will end up with a badly placed screw. A method is described for solving the problem by the addition of a second screw. PMID- 3038234 TI - Internalization of atrial natriuretic factor by AtT-20 corticotropin-secreting cells. AB - Because extended exposure of AtT-20 corticotropin-secreting cells to atrial natriuretic factor (ANF) results in a desensitization of ANF-induced cGMP synthesis, we sought to establish whether pretreatment of AtT-20 cells with the atrial peptide also led to an internalization process. In fact, by coupling an ultrastructural approach to cryoultramicrotomy, ANF-immunoreactivity was detected at both the plasma membrane level and at intracellular sites in AtT-20 cells. Internalization was observed within 5 min at which time labelling was observed in the plasma membrane level, in vacuole-like structures in close proximity to the plasma membrane, in cytoplasmic matrix and sometimes in mitochondria. After 30 min exposure Golgi apparatus, mitochondria and nuclear euchromatin were also labelled. Following 1-4 hr, labelling in other cell compartments, e.g. lysosomal, was increased, while it was reduced in plasma membranes and vacuole-like structures. Secretory granules and endoplasmic reticulum were not labelled throughout the time course. Extraction of a intracellular [125I] ANF from AtT-20 cells following 4 hr incubation suggested that about 90% of the peptide was intact. The data suggest that internalization of ANF may serve to terminate the biological response associated with ANF receptor activation; subcellular distribution of internalized, intact ANF suggests that the peptide may have other, as yet unidentified, intracellular actions. PMID- 3038233 TI - Biochemical and ultrastructural findings in Epstein-Barr virus-transformed lymphoid cell lines from type 1 Gaucher disease. AB - Lymphoid cell lines established by Epstein-Barr Virus (EBV)-transformation of peripheral blood B-lymphocytes from patients affected with type 1 Gaucher disease showed a severe deficiency of glucosylceramide-beta-glucosidase activity (residual activity around 15%-30% of control activity). Ultrastructural investigations showed, in these lymphoid cell lines from type 1 Gaucher disease, the presence of numerous membrane-bound inclusion bodies characteristic of Gaucher cells. PMID- 3038235 TI - In vitro effects of calcium phosphate biomaterials on fibroblastic cell behavior. AB - The effect of synthetic granular hydroxyapatite (HAP) on cultured fibroblastic cells (L929, human bone and gingiva cells) was studied. Phagocytosis of HAP particles and resulting morphological cell changes were demonstrated by microscopic examinations. Cell counts and [3H]thymidine uptake indicated significant increases in cell proliferation and DNA synthesis. These results could account for some of the alterations of the fibroblast behavior induced by changes in intracellular levels of calcium ions released from the material. PMID- 3038236 TI - Electrophysiological effects of Ca antagonists, tetrodotoxin, [Ca]o and [Na]o on myocardium of hibernating chipmunks: possible involvement of Na-Ca exchange mechanism. AB - The electrophysiological performance of myocardium of hibernating chipmunks was investigated in the presence of Ca antagonists and tetrodotoxin, and the effects of high [Ca]o and low [Na]o were examined. The action potential of the preparations was characterized by the low amplitude of the plateau phase (APp). Ca antagonists, nifedipine (10(-6) M) and nitrendipine (2 X 10(-6) M), did not significantly inhibit this APp or the contraction. These nifedipine-insensitive electromechanical responses were completely abolished by an internal Ca release inhibitor, ryanodine. Both increasing [Ca]o and lowering [Na]o, by replacing Na by lithium or choline, also inhibited APp. Tetrodotoxin (10(-5) M) which markedly inhibited the initial rapid phase of the action potential slightly affected APp. These results suggest that the plateau potential of the present preparations is controlled by a process linked to Ca release from internal stores, most likely the Na-Ca exchange mechanism. PMID- 3038237 TI - Effects of cations on binding, in membrane suspensions, of various opioids at mu sites of rabbit cerebellum and kappa-sites of guinea-pig cerebellum. AB - At the mu-sites of rabbit cerebellum, NaCl, LiCl, KCl, choline chloride and MnCl2 were tested for potentiation and inhibition of the binding of several opioids. Naloxone, (-)-bremazocine and diprenorphine are mu-antagonists in pharmacological assays and their binding is potentiated by the lower concentrations and inhibited by the higher concentrations of NaCl. The binding of the agonists [3H]-[D-Ala2, MePhe4, Gly-ol5]enkephalin and [3H]-dihydromorphine is inhibited. MnCl2 potentiates the binding of the agonist [3H]-[D-Ala2, MePhe4, Gly-ol5]enkephalin but not the binding of the antagonists. The thresholds of inhibition and slopes of the dose-response curves for inhibition by MnCl2 and LiCl vary. This finding may indicate that potentiating effects of MnCl2 and LiCl are masked by simultaneous inhibition. At the kappa-sites of guinea-pig cerebellum, NaCl, KCl and MnCl2 inhibit the binding of [3H]-dynorphin A (1-8), [3H]-dynorphin A (1-9), [3H]-(-)-bremazocine, [3H]-tifluadom, and [3H]-diprenorphine. NaCl also causes a small potentiation of the binding of [3H]-diprenorphine, which is a kappa-agonist in the guinea-pig myenteric plexus but a kappa-antagonist in the rabbit vas deferens. The slopes of the inhibitory dose-response curves and the thresholds of inhibition vary with the different ligands. Therefore some potentiating effects may have been masked. The results support the view that NaCl, and perhaps LiCl, but not KCl and choline chloride, potentiate the binding of mu-antagonists but not the binding of mu-agonists. It is not yet possible to decide whether, at the kappa-site, there is a similar differentiation of the binding of agonists and antagonists. PMID- 3038238 TI - The effect of reserpine on sympathetic, purinergic neurotransmission in the isolated mesenteric artery of the dog: a pharmacological study. AB - Electrical transmural stimulation evoked a transient contraction in the isolated mesenteric artery of the dog. This contraction was abolished by guanethidine or tetrodotoxin and was partially inhibited by prazosin. Noradrenaline was competitively antagonized by prazosin. Similarly, in the reserpine-treated artery, electrical transmural stimulation produced a transient contraction which was abolished by guanethidine or tetrodotoxin. However, prazosin failed to inhibit this contraction. The contraction to noradrenaline was not significantly different from the response it produced in control vessels. Tyramine (10(-5) M), which acts on sympathetic nerves to release noradrenaline, evoked a tonic contraction in the untreated artery. This contraction was abolished or markedly attenuated by prazosin or guanethidine. The response was not observed in the reserpine-treated artery, indicating that reserpine had depleted the nerves of noradrenaline. In the control vessel alpha,beta-methylene-ATP produced a transient contraction which was followed by a complete relaxation to the basal level. This contractile response was not significantly different in the presence of guanethidine or prazosin or in the reserpine-treated artery. After desensitization of the vessel to alpha,beta-methylene ATP (5 X 10(-6) M) the prazosin-resistant contractions induced by electrical transmural stimulation were abolished both in reserpine-treated and untreated arteries. Also the contractile responses to ATP and alpha-beta-methylene-ATP were abolished but the responses to tyramine (control vessels), noradrenaline and KCl were not affected. 8 Phenyltheophylline (10(-5) M) showed no inhibitory effect on the contractile responses to electrical transmural stimulation, tyramine, ATP or alpha,beta methylene-ATP. 7. Neuropeptide Y, peptide YY, vasoactive intestinal polypeptide, bombesin and substance P (10-7 and 10-6 M for each peptide) caused no contractile response in the dog mesenteric artery. 8. These experiments provide further evidence that the sympathetic contraction of the isolated mesenteric artery of the dog induced by electrical transmural stimulation consists ofan adrenergic and a purinergic component and that the latter component is mediated through postsynaptic P2- purinoceptors. PMID- 3038240 TI - GABAB-receptor mediated inhibition of potassium-evoked release of endogenous 5 hydroxytryptamine from mouse frontal cortex. AB - The effect of baclofen, the GABAB-agent, on the potassium-evoked release of endogenous 5-hydroxytryptamine (5-HT) from slices of mouse frontal cortex has been investigated. The release of endogenous 5-HT evoked by addition of K+ (35 mM) was inhibited by (+/-)-baclofen in a dose-dependent manner with an IC50 of 0.1 microM. Inhibition of K+-evoked release of 5-HT was produced by (+/-)- and ( )-baclofen but not (+)-baclofen. This action of the (-)-enantiomer was not altered by the presence of the (+)-enantiomer. Addition of GABA (0.1-10 microM) also induced a dose-dependent inhibition of 5-HT release. This effect was neither enhanced by flurazepam (1 microM) nor antagonized by bicuculline (10 microM). The progabide metabolite, 4-[( (4-chlorophenyl) (5-fluoro-2 hydroxyphenyl)methylene]amino)butyric acid (SL75.102) (1 microM) inhibited the K+ evoked release of 5-HT by 61%. These data suggest that baclofen is a potent inhibitor of the K+-evoked release of endogenous 5-HT from the cortex and further indicate that the release of 5-HT may be controlled by a GABAB-receptor located presynaptically. PMID- 3038239 TI - Calcium- and cyclic AMP-dependent chloride secretion in human colonic epithelia. AB - Three stable epithelial cell lines (HCA-7, HCA-7-Col 1 and HCA-7-Col 3) all derived from the same human adenocarcinoma have been cultured on collagen-coated Millipore filters. These epithelial monolayers have been used to record short circuit current (SCC) in response to of secretagogues. Similar monolayers, but grown on plastic dishes, were used for measurements of tissue cyclic AMP. Lysylbradykinin, applied to either side of the monolayers, increased SCC in HCA-7 cells but had little effect on the other two lines. The responses showed rapid desensitization, which could be prevented by cooling to 4 degrees C. Responses to kinin were not significantly attenuated by piroxicam, an inhibitor of cyclo oxygenase. Other secretagogues, vasoactive intestinal polypeptide (VIP) and carbachol also increased SCC in monolayers. The responses to VIP were greatest in HCA-7-Col 1 monolayers while responses were virtually absent in HCA-7-Col 3. A similar profile was seen with carbachol except that responses of HCA-7 and HCA-7 Col 1 monolayers were more equal. With one exception the responses to VIP and carbachol showed sidedness, acting only from the basolateral side. The effects of the secretagogues were inhibited by piretanide, a loop diuretic, applied basolaterally. It is presumed that SCC responses represent electrogenic chloride secretion. Treatment with forskolin increased SCC in HCA-7 and HCA-7-Col 1 monolayers with little effect in HCA-7-Col 3. Nevertheless cyclic AMP levels were elevated most in HCA-7-Col 3 and least in HCA-7-Col 1 monolayers, in reciprocal relationship to the functional response. A23187 increased SCC when applied to HCA 7 and HCA-7-Col 3 monolayers with little effect on HCA-7-Col 1. The differential responses of the three human cell lines provide unique opportunities to discover the functional responsibilities of entities involved in the chloride secretory process. HCA-7-Col 3 cells which generate high levels of cyclic AMP in response to forskolin but which fail to show a substantial chloride secretory response may be a useful model of some disease conditions. PMID- 3038241 TI - Bremazocine differentially antagonizes responses to selective mu and delta opioid receptor agonists in rat hippocampus. AB - The effects of mu, delta and kappa opioid receptor agonists were examined on evoked field potentials in brain slices prepared from rat hippocampus. The effects of the mu-selective opioid peptide [D-Ala2, NMe-Phe4, Met(O)5ol]enkephalin (FK 33-824) and the delta-selective peptide [D-Pen2, D Pen5]enkephalin (DPDPE) were qualitatively and quantitatively similar. Both increased the amplitude of evoked population spike responses when perfused in low nanomolar concentrations in a fashion consistent with what has been previously reported for other opiate agonists such as morphine. The kappa-selective agonists bremazocine and U-50, 488H were without effect upon evoked responses at concentrations as high as 10 microM. Bremazocine, but not U-50, 488H, proved to be an extremely potent antagonist of responses to both mu- and delta- selective agonists. Moreover, bremazocine was considerably more potent in antagonizing responses to FK 33-824 than DPDPE, which supports the hypothesis that FK 33-824 and DPDPE act via different receptors. Thus, although bremazocine is an agonist at kappa receptors, it appears to act as an antagonist at other opioid receptor sites. PMID- 3038242 TI - A comparison of excitatory amino acid antagonists acting at primary afferent C fibres and motoneurones of the isolated spinal cord of the rat. AB - The potency of 7 excitatory amino acid antagonists had been measured at kainate receptors on primary afferent C fibres using isolated dorsal roots from immature rats. Two of the compounds were tested as antagonists of excitant amino acids at motoneurones using isolated hemisected spinal cord preparations. Mean dose-ratios for antagonism of kainate at C fibres, produced by 1 mM antagonist in at least four preparations, ranged from 2.1 +/- 0.5 (mean +/- s.e.mean) with 2-amino-5 phosphonopentanoate (AP5) to 17.6 +/- 2.4 with 1-(p-bromobenzoyl)-piperazine-2, 3 dicarboxylate (BBpzD). The rank order of potency of antagonists at C fibres was similar to that obtained previously for antagonism of kainate at motoneurones. The potency of kynurenate as an antagonist of kainate at C fibres (apparent Kd 70 +/- 4.3 microM (mean +/- s.e.mean), n = 12) was significantly different (P less than 0.005, Wilcoxon rank sum) from its potency at motoneurones (apparent Kd 164 +/- 14 microM, n = 13). Kynurenate also was significantly (P less than 0.025 Wilcoxon rank sum) more potent at antagonizing kainate- than quisqualate (apparent Kd 258 +/- 28 microM, n = 12)-induced depolarization of motoneurones. Kynurenate and BBpzD (0.25, 1.0 and 4.0 mM) were compared as antagonists of N methyl-D-aspartate (NMDA) at motoneurones and the slope of the Gaddum-Schild plot for kynurenate was markedly greater than 1 (2.01 +/- 0.22, 95% confidence limits). A greater than additive antagonism of NMDA-induced depolarizations was produced by combinations of kynurenate with, either AP5, or magnesium ions. 5. The results suggest that the kainate receptor on C fibres may be different from the kainate receptor on motoneurones and that antagonism of NMDA-induced depolarization by kynurenate may not be entirely competitive. PMID- 3038244 TI - Effect of the K+ efflux stimulating vasodilator BRL 34915 on 86Rb+ efflux and spontaneous activity in guinea-pig portal vein. AB - The effect of BRL 34915 on 86Rb+ efflux and myogenic activity was studied simultaneously in guinea-pig portal vein. 86Rb+ was used as a tracer ion for K+. BRL 34915 inhibited myogenic activity with an IC50 value of 12 +/- 2 nM by reducing primarily the frequency of spontaneous contractions. Washout of the substance was followed by hyperreactivity of the vessel. 86Rb+ efflux was slightly reduced by concentrations of BRL 34915 below 100 nM; above 300 nM efflux was increased in a concentration-dependent manner. Above 10 microM BRL 34915, a slow desensitization of the effect on flux was observed during the 10 min application period of the agonist. The Ca2+ entry blocker, isradipine (PN 200 110, 200-500 nM) did not modify BRL 34915-stimulated 86Rb+ efflux at any BRL 34915 concentration tested, indicating that the influx of extracellular Ca2+ through dihydropyridine-sensitive Ca2+ channels is not necessary for this effect. However, by abolishing spontaneous activity, it allowed the 86Rb+ efflux promoting effect of BRL 34915 to be observed at a concentration of 60 nM. The K+ channel blockers tetraethylammonium and 3,4 diaminopyridine inhibited the BRL 34915-induced 86Rb+ efflux with IC50 values of 13 and 3 mM, respectively. Cell permeable derivatives of cyclic AMP and cyclic GMP had no major effect on BRL 34915-induced 86Rb+ flux, indicating that cyclic nucleotide-induced phosphorylation does not play an important modulatory role here. In conclusion, there is an at least 5 fold difference between the concentrations of BRL 34915 necessary to inhibit myogenic activity and those needed to stimulate 86Rb+ efflux. This may be explained by a primary effect of BRL 34915 on the pacemaker cells of the portal vein. explained by a primary effect of BRL 34915 on the pacemaker cells of the portal efflux. This may be PMID- 3038243 TI - Effects of phencyclidine, SKF 10,047 and related psychotomimetic agents on N methyl-D-aspartate receptor mediated synaptic responses in rat hippocampal slices. AB - The effects of representative drugs from three classes of psychotomimetic compounds (arylcyclohexylamines, benzomorphan opioids and dioxolanes) have been examined on synaptic transmission at an identified monosynaptic pathway in rat hippocampal slices. The compounds tested were phencyclidine (PCP) and ketamine, the racemate and isomers of SKF 10,047 (N-allylnormetazocine), and the isomers of dioxadrol (dexoxadrol and levoxadrol). In the absence of added magnesium ions (Mg) in the perfusion medium low frequency stimulation of the Schaffer collateral commissural pathway evoked a burst of population spikes in the CA1 cell body region. The secondary components of this response could be abolished by the selective N-methyl-D-aspartate (NMDA) antagonist D-2-amino-5-phosphonovalerate (APV). PCP (1 microM) or ketamine (10 microM) selectively blocked the secondary components of the synaptic response. The effect of PCP was neither mimicked nor prevented by hexamethonium and atropine, phentolamine and propranolol, or clonidine and was therefore unlikely to involve cholinergic or adrenergic neurotransmitter systems. The sigma opiate, (+/-)-SKF 10,047 (10 microM) also abolished selectively the secondary components of the synaptic response. There was no apparent difference between the potency of the stereoisomers of this compound. The action of (+/-)-SKF 10,047 was not affected by either naloxone or haloperidol, indicating that this effect did not involve opioid receptors or the haloperidol-sensitive sigma site. Dexoxadrol (10 microM), but not levoxadrol (10 microM), also selectively blocked the secondary components of the synaptic response. It is concluded that these psychotomimetic agents can block an NMDA receptor-mediated component of synaptic transmission in the hippocampus and that this effect is mediated by a specific PCP/sigma site. PMID- 3038245 TI - The contributions of mu-, delta- and kappa-opioid receptors to the actions of endogenous opioids on spinal reflexes in the rabbit. AB - Spinal reflexes in the rabbit are suppressed tonically by endogenous opioids. The contributions made to this suppression by mu-, delta- and kappa-opioid receptors have been investigated by studying the actions of a range of opioid antagonists and agonists on reflexes evoked by sural nerve stimulation in the ankle extensor gastrocnemius medialis (g.m.), and in the knee flexor semitendinosus (s.t.). When given at a total dose of 88.5 micrograms kg-1 i.v., either of the universal opioid receptor antagonists (-)-naloxone and (-)-quadazocine enhanced the g.m. response to more than 7 times the pre-drug control values, and the s.t. reflex to 1.5 times controls. The effects of quadazocine were stereospecific. The selective delta antagonist ICI 174864 (3.5 mg kg-1 i.v. total) also augmented the g.m. reflex but only to twice pre-drug controls. The mu-agonists fentanyl (100 micrograms kg-1) and morphine (50 mg kg-1) suppressed both g.m. and s.t. reflex responses to less than half control levels by a naloxone-reversible mechanism. The kappa-agonists bremazocine (50 micrograms kg-1 total), tifluadom (100 micrograms kg-1), ethylketocyclazocine (200 micrograms kg-1) and U50488H (1 mg kg 1) potentiated the g.m. reflex and had variable effects on the s.t. response. Naloxone usually added to the facilitatory actions of these drugs. kappa-Opioid receptor agonists also caused a profound, naloxone-reversible depression of arterial blood pressure. It may be concluded that the endogenous opioid-mediated suppression of spinal reflexes in the rabbit is mediated mainly, if not exclusively, through mu-receptors. There are no known endogenous ligands which are specific for the mu-receptor, so in the present case it seems that selectivity is determined by the receptor population rather than by the ligand. PMID- 3038246 TI - The affinities, potencies and efficacies of some benzodiazepine-receptor agonists, antagonists and inverse-agonists at rat hippocampal GABAA-receptors. AB - The abilities of some benzodiazepine-receptor agonists, antagonists and inverse agonists to modulate the inhibitory potency of the gamma-aminobutyric acid (GABA)A-receptor agonist, isoguvacine, on the CA1 population spike recorded from slices of rat hippocampus, were determined. Concentration-response curves were constructed of the extent to which the benzodiazepine-receptor ligands shifted the isoguvacine concentration-response curve to the left or right. These were compared to their displacement curves of [3H]-Ro15-1788 binding to rat hippocampal membranes under near physiological assay conditions. The above comparisons suggest that the effect on the potency of isoguvacine produced by the benzodiazepine-receptor agonists, diazepam and flunitrazepam, and the partial agonists, Ro16-6028 and Ro17-1812, closely parallels their degree of benzodiazepine-receptor occupancy. Thus, the partial agonists, Ro16-6028 and Ro17 1812, were unable to produce as large a maximum response as the full agonists, diazepam and flunitrazepam. The maximum effects produced by diazepam, flunitrazepam, Ro16-6028, Ro17-1812, the antagonist, propyl-beta-carboline-3 carboxylate, and the inverse agonist, methyl-6, 7-dimethyl-4-ethyl-beta-carboline 3-carboxylate (DMCM), on the potency of isoguvacine in the hippocampal slice corresponded to the change in their affinities produced by the addition of GABA in the radioligand binding studies (GABA-shift). This suggests that the changes in affinity of benzodiazepine-receptor ligands produced by GABAA-receptor activation reflects their ability to modify GABAA-receptor function. The benzodiazepine-receptor antagonists, Ro15-1788 and CGS 8216, had apparent agonist and inverse agonist effects, respectively, on the potency of isoguvacine. These effects occurred at concentrations above those required for saturation of the benzodiazepine-receptor, as labelled by [3H]-Ro15-1788, and were not in agreement with the absence of any effect of GABAA-receptor stimulation in the GABA-shift experiments. This indicates that these events are not mediated by an action at the classical benzodiazepine-receptor site. 6 It is concluded that hippocampal GABAA-receptor function can be allosterically modulated in a manner consistent with the agonist/inverse-agonist model of benzodiazepine-receptor activation, and that compounds exist with varying efficacies throughout this range. PMID- 3038247 TI - Adaptation of the GABAA-receptor complex in rat brain during chronic elevation of GABA by ethanolamine O-sulphate. AB - Slice preparations of rat cuneate nucleus were used for studies on the gamma aminobutyric acid GABAA-receptor complex following chronic and acute pretreatment with GABA-alpha-ketoglutarate aminotransferase (GABA-T) inhibitors. The whole brain GABA concentration was significantly increased 2.9 fold and 2.6 fold following treatment with ethanolamine O-sulphate (EOS, orally) for 15-30 days and 56-64 days, respectively. One hour after a single injection of gamma-acetylenic GABA (GAG) i.p., there was a significant 2.1 fold increase in whole brain GABA. Superfusion of a slice with muscimol or the GABA uptake inhibitor nipecotic acid depolarized the afferent nerve fibres. These effects were potentiated by flurazepam (1 microM) and pentobarbitone (10 microM) and antagonized by picrotoxin (3 microM, 30 microM). Following 15-30 days of EOS-treatment, the depolarization response to muscimol was decreased and that to nipecotic acid increased. These changes were no longer significant by 56-64 days of pretreatment. The acute dose of GAG did not affect the depolarization response to muscimol but increased that to nipecotic acid. The potentiations of muscimol by flurazepam (1 microM) and pentobarbitone (10 microM) were enhanced following chronic EOS treatment (15-64 days). The enhancement of flurazepam was less after 56-64 days than after 15-30 days pretreatment whereas the enhancement of pentobarbitone was similar at both times. Acute GAG treatment had no effect. The potency of picrotoxin as an antagonist of muscimol was reduced following chronic EOS treatment; the enhancement was less after 56-64 days than after 15-30 days pretreatment. Acute GAG treatment caused only a very small reduction in picrotoxin potency. Possible adaptations in the GABAA-receptor complex and its modulation during chronic elevation of brain GABA are discussed. PMID- 3038248 TI - Potentiation of adenosine triphosphate-induced contractile responses of the guinea-pig isolated vas deferens by adenosine monophosphate and adenosine 5' monophosphorothioate. AB - The effects of incubating the guinea-pig isolated vas deferens in the presence of adenine nucleotides (adenosine triphosphate, ATP; adenosine diphosphate, ADP; and adenosine monophosphate, AMP), or in the presence of their phosphorothioate analogues (adenosine 5'-O-(3-thiotriphosphate), ATP gamma S; adenosine 5'-O-(2 thiodiphosphate), ADP beta S; and adenosine 5'-monophosphorothioate, AMP alpha S), on contractile responses to ATP were compared. After challenge with a low (1 microM) or high (300 microM) concentration of ATP to obtain control responses, one vas deferens of a pair was incubated for 5 min with one of the adenine nucleotides, while the contralateral preparation was incubated with the corresponding phosphorothioate analogue. At the conclusion of the incubation the preparations were challenged again with ATP. Incubation with AMP or AMP alpha S resulted in a transient potentiation of responses to 1 microM and 300 microM ATP. The potentiation following incubation with AMP alpha S was larger than that produced by AMP. After incubation with ADP, ADP beta S, ATP and ATP gamma S, responses to 1 microM ATP were decreased, while those to 300 microM ATP were unaffected. Thus, incubation with AMP and AMP alpha S results in potentiation, rather than inhibition, of ATP-induced responses. On the other hand, 5' diphosphate, 5'-triphosphate, 5'-O-(2-thiodiphosphate) and 5'-O-(3 thiotriphosphate) moieties on adenosine have no effect or cause autoinhibition. These results indicate that AMP exerts a potentiating effect on reactivity to exogenous ATP. AMP arising from the enzymatic degradation of ATP might modulate the level of response to ATP released endogenously as a cotransmitter. PMID- 3038250 TI - Technetium pyrophosphate muscle scans in inflammatory muscle disease. AB - Technetium-99M pyrophosphate (TcPYP) nuclear scans of extremities were performed on 15 patients at 10 minutes and 2 hours after isotope injection. Scans were carried out both to confirm the diagnosis of myositis and to direct subsequent muscle biopsy. Five of six patients with clinical features strongly suggestive of inflammatory muscle disease had positive scans. All muscle biopsies performed at areas of increased isotope uptake showed inflammatory muscle disease. All nine patients not suspected of active inflammatory muscle disease had negative scans. Two of these underwent muscle biopsy with negative results. Our observations suggest that TcPYP muscle scans may be useful both to confirm the clinical suspicion of inflammatory muscle disease and in directing the choice of site for muscle biopsy. PMID- 3038251 TI - Milwaukee shoulder--synovial fluid contains no active collagenase. PMID- 3038249 TI - Studies on the adenosine-receptor mediating the augmentation of histamine-induced inositol phospholipid hydrolysis in guinea-pig cerebral cortex. AB - Incubation (45 min) of slices of guinea-pig cerebral cortex with adenosine alone had no significant effect on the accumulation of [3H]-inositol phosphates but enhanced the response to histamine H1-receptor stimulation in a concentration dependent manner. The effect of adenosine on agonist-stimulated inositol phospholipid hydrolysis appeared to be selective for histamine H1-receptor stimulation since it did not augment the phosphoinositide responses to carbachol, noradrenaline, 5-hydroxytryptamine or elevated KCl. The accumulation of [3H] inositol phosphates induced by histamine increased linearly between 5 and 45 min incubation with agonist. However, following the simultaneous addition of histamine and adenosine, there was a marked delay in the appearance of the augmentation produced by adenosine. The augmentation of [3H]-inositol phosphate accumulation was mimicked by a number of adenosine analogues. The rank order of potency was; cyclopentyladenosine greater than R-phenyl-isopropyladenosine 5'-N ethylcarboxamidoadenosine greater than 2-chloroadenosine. This is consistent with the order expected for an adenosine A1-receptor effect but the EC50 values were in the micro- rather than nanomolar range. The response to 2-chloroadenosine was antagonized by the xanthine adenosine-antagonists, cyclopropyltheophylline, 8 phenyltheophylline, 3-isobutyl-1-methylxanthine and theophylline, and the non xanthine alloxazine. PMID- 3038252 TI - Evaluation of hepatic resection for hepatocellular carcinoma in the elderly. AB - The results of 139 hepatic resections in 154 patients with hepatocellular carcinoma (HCC) operated upon in the last 6 years are reported. These 139 Japanese patients were classified into three groups according to age. Group I included 58 patients under 55 years of age, group II 44 aged from 56 to 65 years and group III 37 aged over 66 years. Fewer cases in group III had histological evidence of cirrhosis than in the other two groups. There was a statistically significant marked decline in the preoperative cardiac and pulmonary function in group III patients. However, there was no statistical difference in postoperative complications, mortality and survival rate among the three groups. The results after hepatic resection seemed to depend on pre-operative liver function and histology, and on control of the haemorrhage at operation. The results indicate that liver surgery for elderly patients with hepatocellular carcinoma is justifiable given careful patient selection. PMID- 3038253 TI - Congenital cytomegalovirus infection. PMID- 3038255 TI - Why women are not receiving anti Rh prophylaxis. PMID- 3038254 TI - Converting enzyme inhibition and kidney function in normotensive diabetic patients with persistent microalbuminuria. AB - The effects of a long term reduction in blood pressure on the kidney function of normotensive diabetic patients who had persistent microalbuminuria (30-300 mg albumin/24 hours) were studied in two groups of 10 such patients before and during six months of treatment with either 20 mg enalapril or placebo daily. Treatments were assigned randomly in a double blind fashion. Before treatment both groups had similar clinical characteristics, weight, diet, total glycosylated haemoglobin, median albumin excretion rate (enalapril group 124 mg/24 h, placebo group 81 mg/24 h), and mean arterial pressure (enalapril group 100 (SD 8) mm Hg, placebo group 99 (6) mm Hg). During treatment weight, urinary urea excretion, and total glycosylated haemoglobin remained unchanged. The mean arterial pressure decreased in the enalapril group but not in the placebo group (enalapril group 90 (10) mm Hg, placebo group 98 (8) mm Hg). The median albumin excretion rate also fell in the enalapril group but not in the placebo group (enalapril group 37 mg/24 h, placebo group 183 mg/24 h.) The glomerular filtration rate rose in the enalapril group from 130 (23) ml/min/1.73 m2 to 141 (24) ml/min/1.73 m2, and total renal resistances and fractional albumin clearance decreased while fractional albumin clearance increased in the placebo group. These results show that in patients who have diabetes but not hypertension a reduction in blood pressure by inhibition of converting enzyme for six months can reduce persistent microalbuminuria, perhaps by decreasing the intraglomerular pressure. PMID- 3038256 TI - Deoxyribonucleic acid (DNA) polymorphism of the alpha 1-antitrypsin gene in chronic lung disease. AB - Specific gene probes were used to study restriction fragment length polymorphisms of the human alpha 1-antitrypsin gene. A polymorphism due to loss of a recognition site for the restriction enzyme Taq I was identified in eight of 42 patients with bronchiectasis and nine of 49 patients with pulmonary emphysema, none of whom had alpha 1-antitrypsin deficiency. Among a control group without lung disease the polymorphism was significantly less frequent, being found in only five of 101 apparently healthy blood donors. The deoxyribonucleic acid (DNA) polymorphism was also present in two of 14 unrelated patients with alpha 1 antitrypsin deficiency, indicating a lack of association with any specific alpha 1-antitrypsin protein phenotype. The polymorphism identified in this study may be a new marker for genetic predisposition to chronic lung disease. PMID- 3038258 TI - Characterization of PGE2 inhibition of corticotropin releasing factor-mediated ACTH release. AB - The role of prostaglandin E2 (PGE2) on the mechanism of corticotropin releasing factor (CRF) induced adrenocorticotropin (ACTH) release was studied in primary rat pituitary cell culture. The continuous incubation of pituitary cells with PGE2 inhibited CRF-stimulated ACTH with an ED50 of 1.2 X 10(-9) M PGE2. PGE2, however, did not alter the spontaneous release of ACTH. PGE (10(-8) M) significantly decreased 10(-10) M, 10(-9) M, and 10(-8) M CRF-mediated ACTH release by 42%, 47%, and 31% of total CRF stimulated ACTH release. Time course experiments demonstrated a PGE2-induced inhibition by 20 min of CRF incubation which continued for 3 h. After a 2-h incubation with PGE2, the wash-out of PGE2 from the culture medium just prior to the addition of CRF eliminated the inhibitory activity of PGE2. PGE2 decreased the amount of CRF-stimulated ACTH from cells incubated with cycloheximide (P less than 0.01). The inhibitory activity of PGE2 (10(-8) M) on CRF-stimulated ACTH was unaltered by the addition of 3 mM or 7 mM CaCl2 to the standard culture media (1.6 mM CaCl2). The inhibition of CRF-induced ACTH release by maximal inhibitory concentrations of PGE2 (10(-7) M) and cortisol (5 X 10(-7) M) were not additive. In conclusion, PGE2 may play an important role in modulating pituitary ACTH release. Its inhibitory activity occurs by 20 min of CRF incubation, is in part independent of protein synthesis, requires the continued presence of PGE2, is not reversed with CaCl2, and is not additive with the inhibitory activity of cortisol. PMID- 3038259 TI - Conduction properties of single nerve fibers in developing rat spinal nerve roots. AB - We have examined conduction properties and distribution of membrane currents in ventral spinal root axons of rats 12-47 days of age. Internodal length of the largest myelinated fibers increases at a steady 17-19 micron/day during this period as internodal conduction time decreases. Despite the rapid remodelling of fiber dimensions during the first few weeks of postnatal life, potassium channels are continually excluded from participation in action potential generation at the axon membrane of most nodes of Ranvier. PMID- 3038257 TI - Cough associated with captopril and enalapril. AB - Thirty three reports of cough associated with captopril and 26 associated with enalapril received by the New Zealand intensive medicines monitoring programme were reviewed. The programme is a specialised part of the New Zealand postmarketing surveillance system. Review of these reports showed that the cough was an adverse reaction to the drugs, occurred even with low dose treatment, and was severe enough to warrant withdrawal of the drugs in most of the cases reported. A significant sex difference was shown, with women predominating. The reaction seemed to be a greater problem with enalapril, and in seven patients it occurred with both captopril and enalapril. Withdrawal of treatment resulted in rapid recovery, and no long term effects were shown. The pathogenesis of the reaction is unknown, but possible mediators include bradykinin and prostaglandins. PMID- 3038260 TI - GABA-receptor stimulation enhances norepinephrine-induced polyphosphoinositide metabolism in rat hippocampal slices. AB - The effect of gamma-aminobutyric acid (GABA)receptor stimulation on norepinephrine (NE)-induced metabolism of polyphosphoinositides (PIPs) was studied in rat hippocampal slices. Inositol phosphates (IPs), PIPs, and phosphatidic acid were measured. NE induced formation of IPs and phosphatidic acid in a dose-dependent manner with an EC50 of 4.5 microM. GABA, 3 aminopropanesulphonic acid (3APS) and muscimol did not affect PIPs breakdown, but they strongly increased (greater than 100%) PIPs metabolism induced by 1 microM NE. Their action was antagonized by bicuculline (10 microM). We discuss the implications of these findings in hippocampal neurotransmission. PMID- 3038261 TI - Benzodiazepine receptors in the human cerebellar cortex: a quantitative autoradiographic and pharmacological study demonstrating the predominance of type I receptors. AB - The anatomical localization of benzodiazepine receptors in the human cerebellar cortex was studied using quantitative autoradiography following in vitro labelling of cryostat sections with [3H]flunitrazepam ([3H]FNZ), and the pharmacology of these receptors has been characterized by computerized, non linear least squares regression analysis of [3H]FNZ displacement by FNZ, CL218,872 and ethyl beta-carboline-3-carboxylate (ECC) binding to membranes. The autoradiograms demonstrated that benzodiazepine receptors were present throughout all layers of the human cerebellar cortex; high concentrations of receptors were present in the molecular layer, moderate concentrations were present in the granular layer and a much lower density of receptors was seen in the intervening Purkinje cell layer. The pharmacological studies indicated that the human cerebellar cortex contained a high concentration of homogeneous benzodiazepine receptors which have high affinity for FNZ, ECC and CL218,872, i.e. type I sites. PMID- 3038262 TI - Cerebral phosphoinositide, triacylglycerol and energy metabolism during sustained seizures induced by bicuculline. AB - In ventilated rats, levels of phosphatidylinositol (PI), phosphatidylinositol 4 phosphate (PIP), phosphatidylinositol 4,5-bisphosphate (PIP2), diacylglycerol (DAG), triacylglycerol (TAG), free fatty acids (FFA) and phosphatidic acid, as well as their fatty acid contents, were measured in forebrain tissue after 1, 20 and 60 min of seizures induced by bicuculline. Cerebral energy state was also measured. PI decreased progressively throughout 60 min of seizures, whereas the levels of PIP and PIP2 did not change. DAG increased modestly and persistently. FFA increased markedly during the early seizure period, but decreased later. Following an initial drop, TAG rose above control. Phosphatidic acid did not change. The levels of ATP and energy charge potential decreased slightly and lactate accumulated. Stearic acid (18:0) and arachidonic acid (20:4) primarily accounted for the changes in the levels of the lipids. At the onset of seizures, the decrease of 18.0 and 20:4 in PI occurred in parallel with an enrichment of these fatty acids in FFA and DAG. Despite the fact that the losses of 18:0 and 20:4 from PI were quantitatively similar to each other at all times examined, the increase in free 18:0 was much larger than the increase in free 20:4 at 20 min of seizures. Concurrently there was a rise of 20:4 in TAG. As the FFA levels declined thereafter, 20:4 and docosahexaenoate (22:6) in TAG continued to increase. The results are consistent with the view that seizure activity stimulates the hydrolytic breakdown of brain phosphoinositides--the pathway catalyzed by phosphodiesterase of the phospholipase C type followed by lipases, and probably the pathway catabolized by phospholipases A as well. Preferential incorporation of polyunsaturated fatty acids into TAG-acyl residues may represent a mechanism to reduce the level of their free forms when the latter are produced in large amounts. PMID- 3038263 TI - A cholinergic projection to the rat substantia nigra from the pedunculopontine tegmental nucleus. AB - This study demonstrates the pedunculopontine tegmental nucleus (PPN) to be the source of a major cholinergic projection to the rat substantia nigra (SN). Neurons of the PPN were double-labeled utilizing choline acetyltransferase immunocytochemistry combined with retrograde transport of horseradish peroxidase (HRP). The cholinergic projection to the SN originates from neurons located in predominately the ipsilateral and rostral portions of the PPN. Other cholinergic neurons that were also retrogradely labeled with HRP were located in the caudal PPN and in the lateral dorsal tegmental nucleus. A non-cholinergic projection from this region was also identified. These findings indicate that the PPN may be topographically organized with respect to its efferent projections. In addition, this study provides evidence for an extrinsic source of acetylcholine to the basal ganglia and implicates the PPN as a source of potentially significant influence over basal ganglia function. PMID- 3038264 TI - Localization of neurotensin receptors in the forebrain of the common marmoset and the effects of treatment with the neurotoxin 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine. AB - The distribution of neurotensin binding sites was mapped in the brain of the common marmoset using [3H]neurotensin as the ligand. Autoradiographic techniques show that the density of receptors is particularly high in the substantia nigra, ventral tegmental area, olfactory tubercle and cerebral cortex and that the distribution of neurotensin receptors in the cerebral cortex and striatum is heterogenous. In the cerebral cortex neurotensin receptors are concentrated in layers I, II, III and V, whilst receptor density is generally less in all layers of middle temporal cortex. Striatal neurotensin receptors conformed to a striosomal distribution as defined by acetylcholinesterase staining with the highest density of binding sites in the matrix. The neurotoxin 1-methyl-4-phenyl 1,2,3,6-tetrahydropyridine selectively destroyed over 80% of the dopaminergic neurons of the marmoset substantia nigra and almost 60% of those in the adjacent ventral tegmental area. The subsequent loss of a large proportion of neurotensin receptors from marmoset substantia nigra and striatum suggests their presence on the dopaminergic nigrostriatal pathway. PMID- 3038265 TI - Glycoprotein pattern in human brain tumors studied using lectin binding after sodium dodecyl sulfate-gel electrophoresis and protein blotting. AB - The immunoblotting technique was used to study the glycoproteins in human brain tumor samples including astrocytoma, glioblastoma, meningioma and oligodendroglioma, as well as in normal human brain. Glycoproteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis, electrophoretically transferred to nitrocellulose membrane and characterized, using binding with 11 different lectins. Tumor-associated glycoproteins were found using the lectins peanut agglutinin (PNA), soybean agglutinin, Limulus polyhemus, Lotus tetragonolobus, Ricinus communis 1, (RCA-1) and wheat germ agglutinin (WGA). Their molecular masses ranged from 50 to 180 kDa. Several of them were common to the 3 types of tumors: astrocytomas, oligodendrogliomas and meningiomas. PNA, RCA 1 and WGA were the 3 most feasible lectins with regard to tumor specificity, simplicity and reproducibility. PMID- 3038266 TI - Mechanism of action of lithium on acetylcholine receptor metabolism in skeletal muscle. AB - Changes in the levels of cations within skeletal muscle are thought to mediate the neural regulation of turnover of extrajunctional acetylcholine receptors (AChRs). We have used lithium as a probe of these cation influences because of its resemblance to calcium and other ions. In the present experiments we studied the mechanism of action of lithium on AChR metabolism in cultured mammalian skeletal muscle. We measured the effects of lithium on AChR turnover (using [125I]alpha-bungarotoxin binding), and evaluated the resemblance of lithium and calcium in producing their effects on AChR metabolism. Our results provide insight into the mechanisms of action of lithium and the cellular processes controlling AChR metabolism in muscle. Lithium reduces the number of AChRs in skeletal muscle in vitro to a degree similar to that which we previously reported in vivo. Lithium appears to enter cells via both sodium and calcium channels. It then produces its effect on levels of AChRs primarily by selectively reducing AChR synthesis and insertion into the surface membrane. Lithium induces this change in AChR metabolism in a manner resembling neural and calcium-mediated effects on AChRs. Phosphoinositide pathways may be involved in the lithium induced effects. Further analysis of the effects of lithium on AChR turnover should provide new information about the mechanisms underlying the cellular control of receptor metabolism. PMID- 3038267 TI - The development and decay of kindling-induced increases in paired-pulse depression in the dentate gyrus. AB - Epileptogenic stimulation (kindling) leads to an increase in recurrent inhibition in the dentate gyrus, possibly due to an increase in benzodiazepine receptors. A second, late inhibitory component also potentiates as a result of kindling. In the present experiments, the time course of the development and decay of this kindling-induced increase in inhibition was studied. Rats were kindled by stimulation of the perforant path or by direct stimulation of the dentate gyrus. Paired-pulse stimulation was applied to the perforant path and field potential measures were taken within the dentate gyrus. These procedures allowed the monitoring of inhibitory events in chronic preparations over prolonged periods. Input/output measures of the baseline responses were also monitored during the course of, and after the completion of kindling. The increase in the early component (about 20-50+ ms) of paired-pulse depression was seen after the first kindling stimulation. The increase in the late component of depression (about 100 500+ ms) did not develop until after about 10 stimulations had been delivered. The late component then decayed more rapidly than the early component after the completion of kindling. The baseline response also showed some indication of depression, particularly in the dentate gyrus kindled group, raising the possibility that feedforward inhibition had also been potentiated. PMID- 3038268 TI - An in vitro autoradiographic analysis of mu and delta opioid binding in the hippocampal formation of kindled rats. AB - Recent studies have shown that opioid peptide levels are altered in hippocampal formation of kindled animals. We therefore studied the distributions of mu and delta opioid binding sites in hippocampal formation of kindled and control rats using quantitative in vitro autoradiography. Animals received daily stimulations of the amygdala until they experienced 3 class 5 seizures. Paired control animals underwent implantation of electrodes but were not stimulated. Mu binding sites were labeled with 125I-FK-33824. Twenty-four hours after the last kindled seizure, mu binding was decreased by 32% in stratum pyramidale of CA1 and stratum radiatum of CA2 and by 17-27% throughout most of the rest of CA1, CA2, and CA3. Few, if any, differences were seen between kindled and control animals at 7 or 28 days after the last kindled seizure. Delta binding sites were labeled with 125I [D-Ala2,D-Leu5]enkephalin in the presence of the morphiceptin analog PL-032. Twenty-four hours after the last kindled seizure, delta binding was decreased only in stratum moleculare of the dentate gyrus. Seven days after the last kindled seizure, delta binding was decreased by 11-17% throughout CA1, CA3, and the dentate gyrus. At 28 days after the last seizure, however, no differences were found between kindled and control animals. Since the decreases in mu and delta opioid binding are transient, they are unlikely to be the molecular basis of the permanent kindling phenomenon. Rather, these changes in opioid binding may represent responses to repeated seizures. PMID- 3038269 TI - Responses of ventromedial hypothalamic neurons in vitro to norepinephrine: dependence on dose and receptor type. AB - Application of norepinephrine (NE) at 12.5 microM in the bath surrounding hypothalamic slices from ovariectomized rats could evoke excitation, inhibition, or biphasic inhibition-excitation from single neurons in the ventromedial nucleus. Whether the rats were treated with estrogen or not did not alter the distribution of the type of neuronal responses to NE in vitro. Altering the composition of the bathing solution to achieve synaptic blockade did not abolish or alter the type of responses, indicating that all these types of NE responses, including both phases of the biphasic response, were mediated by postsynaptic receptors. Experiments with varying doses of NE showed that the inhibitory response could be evoked at doses lower than those required to evoke the excitatory response. The effective dose for 50% of the responsive neurons (ED50) was lower than 1.25 microM for inhibitions and higher than 5 microM for excitations. Using specific adrenergic receptor agonists and antagonists, it was found that the excitation and the inhibition were mediated, primarily, by alpha 1 and alpha 2-receptors, respectively. beta-Receptors played only a minor role, but might be related to both excitation and inhibition. Study with adrenergic agents further revealed that different types of adrenergic receptors co-localized not only in neurons showing the biphasic response, but also in a major portion of neurons showing monophasic excitation or inhibition. Because of the co localization and the differential sensitivities to NE, alteration of the dose of NE or the ratio of excitatory/inhibitor receptors co-localized on a neuron should be able to reverse the type of a neuronal response to NE. PMID- 3038271 TI - Fatigue of lateral rectus and retractor bulbi motor units in cat. AB - The fatigue properties of lateral rectus, retractor bulbi and split lateral rectus-retractor bulbi motor units were studied in the cat. Lateral rectus motor units showed a range in resistance to fatigue while retractor bulbi motor units were all fatigable. Within the abducens nucleus, axons of split lateral rectus retractor bulbi motor units are found. These motor units are unique in that one motoneuron projects to two separate muscles. Split motor units were studied to determine if both the lateral rectus and retractor bulbi muscle fibers of split units would show uniform fatigue properties. The results showed that the fatigue resistance of the separate muscle components of these motor units are different, suggesting that the muscle fibers of a motor unit may be physiologically dissimilar. PMID- 3038270 TI - Lithium preincubation stimulates the potassium-induced release of cholecystokinin from slices of cerebral cortex and caudate-putamen incubated in vitro. AB - Potassium-evoked cholecystokinin (CCK) release from slices of caudate-putamen and cerebral cortex, but not hippocampus incubated in vitro was increased by 152-175% by preincubation for 40 min with 10 mM lithium. These results and previous studies suggest that although different physiological agents regulate CCK release in these brain regions, these agents may share a common intracellular mediator which may be a product of inositol phospholipid turnover. PMID- 3038272 TI - Central versus peripheral mediation of responses to adenosine receptor agonists: evidence against a central mode of action. AB - Although adenosine and its analogs have potent effects on the peripheral and central nervous systems, the actions of systemically administered adenosine analogs on the central nervous system are poorly understood. We have previously shown that adenosine analogs depress hippocampal-evoked responses (fEPSPs) in a brain slice preparation, and in the present study, we observed that local pressure ejection of adenosine or R-phenylisopropyladenosine (R-PIA) also reduced the fEPSP recorded in situ in a theophylline-reversible manner. However, systemic administration of R-PIA in up to 1000 times the behaviorally active dose failed to attenuate the fEPSP amplitude. Similar observations were made with systemic injections of 2-chloroadenosine and N-ethylcarboxamidoadenosine, both of which are active in behavioral studies following intraperitoneal or intracerebroventricular injection. Autoradiography showed systemically injected [3H]PIA did not enter the brain in significant concentrations, and this observation was confirmed using microdialysis brain perfusion. These results suggest that systemically administered adenosine analogs do not affect synaptic neurotransmission in the hippocampus because they fail to reach the appropriate receptors. PMID- 3038273 TI - Effects of low calcium and inhibition of calcium-activated neutral protease (CANP) on mature nerve terminal structure in the rat sternocostalis muscle. AB - The possible role of calcium and a calcium-activated neutral protease (CANP) in the reorganisation of mature mammalian neuromuscular junctions was studied in the sternocostalis muscle in rats. After the well-documented loss of polyneuronal innervation has occurred, the remaining single mature nerve ending continues to change its terminal branching pattern by gradually becoming more complex. Reducing local calcium concentrations by the chelating agent BAPTA or inhibiting CANP by local application of an inhibitor, Leupeptin, resulted in the endings becoming more complex in appearance when examined after 6 or 7 days. It is concluded that calcium and CANP are important in the remodelling of mature neuromuscular junctions. PMID- 3038274 TI - Dolichol-linked glycoprotein synthesis in developing mammalian brain: maturational changes of the N-acetylglucosaminylphosphotransferase. AB - The enzyme UDP-N-acetylglucosamine:dolichyl phosphate, N-acetylglucosamine-1 phosphate transferase (GlcNAc-1-P transferase), the first committed step in the dolichol-linked oligosaccharide pathway for glycoprotein biosynthesis, has been studied in developing rat brain. The enzyme was shown to be localized in microsomes, particularly heavy microsomes, and to be activated by Mg2+ and inhibited by tunicamycin. Study of the enzyme with brain development demonstrated two prominent findings. First, the accentuation of enzymatic activity caused by addition of a saturating concentration of dolichyl phosphate was greater in brain of older (3-4 weeks of age and subsequently) animals (25-fold) than in brain of younger (less than two weeks of age) animals (10-fold). This difference suggests that dolichyl phosphate may be limiting for GlcNAc-1-P transferase activity in endoplasmic reticulum of the older animals. Second, a marked (3.5-fold) increase in activity occurred over a discrete time period (3-4 weeks of postnatal life) during brain development. That this increase reflected an increase in enzyme amount rather than in catalytic efficiency was suggested by kinetic studies. Coupled with our previous demonstrations of increases in brain dolichol, dolichol kinase activity, and dolichyl phosphate levels during approximately the same developmental period (Sakakihara, Y. and Volpe, J.J., Dev. Brain Res., 14 (1984) 225-262; Volpe, J.J. et al., Dev. Brain Res., in press), the data suggest a temporally discrete period of activation of the dolichol-linked pathway to glycoproteins. Whether the pathway is regulated coordinately or sequentially is a fertile topic for future study. PMID- 3038275 TI - Vascularity and cytochrome oxidase distribution in the occipital cortex in MAM Ac induced micrencephaly. AB - The laminar distributions of cytochrome oxidase activity and vascularization of the occipital cortex of animals made micrencephalic by exposure to MAM Ac on E14 or E16 were examined and compared to control animals. Despite severe disruptions of the normal cytoarchitecture in MAM Ac exposed animals, the laminar distributions of vascular profiles, branch points and cytochrome oxidase activity in micrencephalic animals were similar to those in control animals. PMID- 3038277 TI - Digestion of starch and fibre carbohydrates in peas by adult cockerels. AB - The digestion of starch and fibre by adult cockerels fed peas which were whole, ground, autoclaved, heated, dehulled, cooked or supplemented with a cellulose degrading enzyme was studied. The starch in ground peas that had been autoclaved, heated or dehulled was slightly, though not significantly, better digested than the starch in peas that had been ground only. Feeding peas whole greatly reduced starch digestibility. Cooking failed to improve starch digestibility as retrograded starch produced in the cooking process was not digested. The more finely ground the foodstuff, the better the in vitro enzymatic hydrolysis of starch. Starch digestibility correlated well (r2 = 0.80) with the true metabolisable energy values obtained for the different pea treatments. In experiment 1 cockerels digested on average 0.22 of the pea fibre from the different forms of peas. A significant decrease in xylose digestion was observed when birds were fed dehulled peas, birds excreting more xylose than they ingested. In experiment 2, cockerels digested on average 0.38 of the pea fibre from different forms of peas. A reduction in fibre digestion was observed when birds were fed cooked peas as a consequence of a decreased digestibility of all monosaccharide residues. A slight increase in fibre digestion was observed when peas were augmented with 'cellulase', entirely because of an increase in xylose digestion. PMID- 3038276 TI - Compartmentalization of the embryonic striatum after intraocular transplantation. AB - In order to assess the intrinsic potential of the isolated embryonic striatum to develop its adult patch and matrix compartments, embryonic day 16 striata were transplanted into the anterior eye chamber of adult host rats. After 2-12 weeks of survival the transplants showed heterogeneous and in the majority of cases complementary distributions of opiate receptor binding and acetylcholinesterase staining, which mark the patch and matrix compartments of the adult striatum, respectively. The complementarity of patch and matrix markers in the transplants shows that the transplants do compartmentalize. However, the density of the markers in the transplants did not reach the levels seen in the adult striatum. The results suggest that the commitment of cells to a striatal compartment is a very early event in the embryonic development of the forebrain. PMID- 3038278 TI - [Demonstration and topographical distribution of LHRH receptors in the central nervous system in the normal and castrated male rat]. AB - The topographical distribution of [125I]-LHRH binding sites was studied on brain sections of adult male rat by quantitative autoradiography. High density of sites was observed in the hippocampus, amygdala and entorhinal cortex (4-7 fmol of LHRH bound/mg protein). Lower density of sites was observed in the septum and frontal cortex. The receptor density was not significantly modified at day 5 following castration. Under the same conditions the pituitary receptors were significantly increased. The presence of specific LHRH binding sites in the limbic system may explain the behavioural effect observed following intracerebroventricular injection of LHRH. However, their functions under physiological conditions remain to be elucidated. PMID- 3038279 TI - Imaging mediastinal tumors. PMID- 3038281 TI - Formation of octacalcium phosphate and subsequent transformation to hydroxyapatite at low supersaturation: a model for cartilage calcification. AB - By means of X-ray powder diffraction (XRD), octacalcium phosphate (OCP, Ca8H2(PO4)6 X 5H2O, Ca/P = 1.33) has been shown to be a metastable precursor of hydroxyapatite (HA, Ca10(PO4)6(OH)2, Ca/P = 1.67) during de novo HA formation in aqueous solutions containing [CaCl2] = 1.0-2.0 mM and [Na2HPO4] less than or equal to 3 mM, kept at 37 degrees C, 300 mosM and stirred gently at 150 rpm. At the precipitation boundary with initial pH = 7.4, OCP is stable for at least 24 hours when [Ca] = 1.0 mM, and is less stable when [Ca] = 2.0 mM, transforming into a mixture of OCP and HA within 24 hours. Almost complete transformation to HA within 24 hours takes place with high [Ca] and high [Pi]. Statistical analysis of the pH 7.4 precipitation boundary data supports the XRD findings: although [Ca][Pi] values vary significantly (P less than 0.001) with [Ca] (2.70 +/- 0.05 mM2 for [Ca] = 1.0 mM and 2.00 +/- 0.10 mM2 for [Ca] = 2.0 mM), [Ca]1.33[Pi] values do not vary with [Ca] suggesting that the initial precipitation process is 6(1.33 Ca + Pi)----OCP. With initial pH = 7.6, a different precipitation boundary with lower [Ca][Pi] values has been determined. These findings strongly support the fact that rat epiphyseal cartilage fluid which has [CaUF][PiUF] = 2.6 mM2 (UF = ultrafiltrate) and pH 7.6 [2] should be able to support de novo calcification. PMID- 3038280 TI - Transmission electron microscopy of lattice planes in human alveolar bone apatite crystals. AB - Periodic fringes corresponding to six different lattice planes have been observed in apatite crystals of human normal alveolar bone by transmission electron microscopy. Three of these sets of fringes have spacings less than 3.5 A corresponding to the Scherzer resolution of the microscope used. The (0002) lattice plane of hydroxyapatite of 3.4 A d-spacings, the (2111) lattice plane with a d-spacing of 2.81 A, and the (3030) lattice plane with a d-spacing of 2.72 A have been identified. The (0002) and (2121) lattice planes have been observed for the first time in bone microcrystals. Some of the crystals studied were characterized by a mean width/thickness ratio of 6.91, typical of platelike habit, whereas observations of crystals aligned along the (1210) and (1211) directions showed a needlelike habit. The mean length of the bone apatite crystals was 470 A. A dark line similar to the one observed in enamel and dentine crystals was also seen. The bone microcrystals observed have shown a high sensitivity to beam damage. PMID- 3038282 TI - Structure-activity relationships (SAR) of hydroxyapatite-binding molecules. AB - Carboxyl-containing molecules can bind to HA with sufficient affinity to prevent the uptake of AZ by this mineral. Structural features that favor binding can be discerned. Isolated carboxyls are inactive; adjacent dicarboxylic functions are marginally active. Crowding of oxygens in the form of vicinal carboxyls or hydroxyls contributes the property of HA binding ability. This property exhibits gradation in magnitude and the variation can be correlated with structural features of the carboxyl-containing molecule. Phospho functions are generally more powerful contributors to HA binding property than are carboxyls. PMID- 3038283 TI - Increased incidence of isoproterenol-induced ventricular fibrillation in aging rats. AB - Prolonged beta-adrenergic stimulation obtained by subcutaneous injection of isoproterenol in unanesthetized, unrestrained rats elicited ventricular fibrillation in approximately 80% of animals at 10-12 months of age. Ventricular fibrillation failed to occur in 1-month-old rats and involved only 12% of rats at 2 months. Senescence appeared not to increase the frequency of ventricular fibrillation since a similar incidence was seen in rats at 10-12 and 19-21 months. In all instances, ventricular fibrillation was preceded by ECG changes consistent with acute subendocardial ischemia. To evaluate whether acute beta adrenergic stimulation elicits comparable cardiovascular effects in animals of different age, a dose-response curve to intravenous injection of isoproterenol was performed in anesthetized rats. Changes in heart rate, systemic arterial pressure, left ventricular pressure, and dP/dt were not different among animal groups. It was concluded that the arrhythmogenic potential of isoproterenol may not be related to differences in cardiac beta-receptor sensitivity with age as suggested by the comparable changes in the inotropic and chronotropic actions of isoproterenol in the animal groups studied. PMID- 3038284 TI - Enzymatic formation of angiotensins II and III in the hindlimb circulation of dogs. AB - Experiments were performed in 14 anesthetized dogs to (1) to determine if the reductions in hindlimb blood flow produced by [des-Asp1] angiotensin I were due to its local enzymatic (kininase II) conversion to angiotensin III and (2) to quantitate the extent of conversion of angiotensin I to angiotensin II and of [des-Asp1] angiotensin I to angiotensin III in the hindlimb circulation. Graded doses of these peptides were administered as bolus injections directly into the left external iliac artery while measuring flow in this artery electromagnetically. Dose-response relationships were determined before and during the inhibition of kininase II activity with captopril or antagonism of angiotensin receptor sites with [Ile7] angiotensin III. Captopril inhibited the vasoconstrictor responses to angiotensin I and [des-Asp1] angiotensin I, but did not affect the responses to angiotensins II or III, or norepinephrine. [Ile7] angiotensin III inhibited the vasoconstrictor responses to all four angiotensin peptides but did not alter the responses to norepinephrine. These findings indicate that the hindlimb vasoconstrictor responses to [des-Asp1] angiotensin I were due to the local formation of angiotensin III. The extent of conversion of [des-Asp1] angiotensin I to angiotensin III that occurred in one transit through the hindlimb arterial circulation was estimated to be 36.7%, which was not different from the estimated 36.4% conversion of angiotensin I to angiotensin II. We conclude that angiotensin I and [des-Asp1] angiotensin I are converted to their respective vasoactive forms (angiotensins II and III) to a similar extent in the hindlimb circulation via the action of kininase II. PMID- 3038285 TI - The release of catecholamines from the adrenal medulla and its modulation by alpha 2-adrenoceptors in the anaesthetized dog. AB - The adrenal nerve of anaesthetized and vagotomized dogs was electrically stimulated (10 V pulses of 2 ms duration for 10 min) at frequencies of 1, 3, 10, and 25 Hz. There was a correlation between the frequency of stimulation and the plasma concentrations of epinephrine, norepinephrine, and dopamine in the adrenal vein, mainly after the 1st min of stimulation and the maximal concentration was reached sooner with higher frequencies of stimulation. Moreover, the relative percentage of catecholamines released in response to the electrical stimulation was not changed by the frequency of stimulation. To test the hypothesis that a local negative feedback mechanism mediated by alpha 2-adrenoceptors exists in the adrenal medulla, the effects of the systemic administration of clonidine (alpha 2 antagonist) on the concentrations of catecholamines in the adrenal vein were evaluated during the electrical stimulation of the adrenal nerve (5 V pulses of 2 ms duration for 3 min) at 3 Hz. Moreover, the effects of the systemic injections of more specific alpha 2-agonist and antagonist (oxymetazoline and idazoxan) were tested on the release of catecholamines in the adrenal vein in response to electrical stimulation of the splanchnic nerve at 1 and 3 Hz frequencies. The injection of 0.5 mg/kg of yohimbine caused a significant increase in the concentrations of epinephrine and norepinephrine in the adrenal vein induced by the electrical stimulation of the adrenal nerve and the injection of 15 micrograms/kg of clonidine had no effects.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3038286 TI - Beta-adrenergic regulation of cardiac myosin. AB - Calcium-independent regulation of the contractile proteins of cardiac muscle has been studied using hyperpermeable cells from rat ventricles and sections of quickly frozen rat hearts. These preparations have been used to study maximum calcium-activated force, myosin ATPase activity, and the maximum velocity of unloaded shortening. Beta-adrenergic activity increases the amount of force and the ATPase activity in accordance with the concentration of the V1 isozyme of myosin. V3 activity is decreased at the same time. In tissues containing only V1, there is no change in maximum velocity in response to beta-adrenergic stimulation. These results indicate that beta-adrenergic stimulation recruits V1 force generators and probably regulates a transition between a calcium unresponsive and a calcium responsive force generator. PMID- 3038287 TI - The pH dependence of the contractile response of fatigued skeletal muscle. AB - Following a period of intense repetitive stimulation (e.g., brief tetanic stimuli every second for 3 min), muscle isometric tension development is reduced by about 80%. This suppression is reversible at a high external pH (8.0) with a half time of 15-20 min, but if the external pH is low (6.4) or the buffer concentration is low, recovery is prevented. Inhibition of recovery is associated with a slowed rate of lactate loss, which may suggest that intracellular lactacidosis is the cause of the inhibition. Alternatively, a low external pH may affect recovery from fatigue quite independently of its effect on lactate efflux. The possibility that surface membrane properties are changed by fatigue in a pH-dependent fashion was examined by measuring the cable properties and action potentials of fatigued fibres at different external pH values. A low external pH during recovery from fatigue was shown to result in a prolonged membrane depolarization of 10-12 mV, an increased transmembrane resistance, and a prolonged action potential. At a high external pH transmembrane resistance is lowered by fatigue, the depolarization lasts only about 10-15 min, and there is a smaller effect on the action potential. While the fatigued fibre membrane does show a changed response that is dependent on external pH, it is not clear that this could be related to the suppression of contraction. Direct measurements of intracellular pH show a fall of about 0.4 to 0.5 pH units in the surface fibres following fatigue. This results from the lactic acid generated during activity. It is now clear that lactate crosses the membrane in association with protons and at least part of this flux is mediated by a specific carrier mechanism. Efflux is limited by the transmembrane pH gradient, which in turn depends on the extracellular buffer concentration in the diffusion limited space around the fibres. Intracellular lactacidosis in resting muscles can be generated by a reversal of the normal flux. Fibres can be loaded with lactate (L) by increasing the extracellular [H+][L-] product with a resultant fall in intracellular pH. Lactate loads similar to those seen in fatigued muscle simulate some but not all of the responses seen in the postfatigue state. The twitch is prolonged with a slow relaxation phase, an increased time to peak tension but with an increase in peak tension. The effects are reversible but usually result in a reduced contractile response following the washout.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3038288 TI - Speculations on disease states induced by excitatory amino acids. AB - There is much current interest in excitatory amino acids and their receptors because of their postulated involvement in several disorders of the nervous system. They function as neurotransmitters, but can act as neurotoxins in some situations. They have been implicated in the pathogenesis of cerebral hypoxic/ischemic and hypoglycemic damage, in epilepsy, in some degenerative diseases, and in some forms of neurotoxin-induced cerebral dysfunction. These diseases may reflect abnormality in a system which has evolved to provide synaptic plasticity essential for learning and memory. The purpose of this paper is to explore the ramifications of such a hypothesis. PMID- 3038289 TI - Peripheral neurological complications of aortoiliac vascular disease. AB - Six patients with an aortoiliac vascular disease and a peripheral neurological deficit are presented. Clinical and electromyographic findings revealed lumbosacral plexus, sciatic and femoral nerve lesions. A correlation is made between the level of the vascular lesion (aortic, aortoiliac or distally) and the type of peripheral nerve deficit observed. In a patient complaining of pain, weakness, or numbness in a leg, the differential diagnosis should include aortoiliac vascular disease. The peripheral neurological symptoms may be the initial manifestation of the vascular disease or may appear in the early post operative period. PMID- 3038290 TI - Comparative properties of feline coronaviruses in vitro. AB - Two feline coronaviruses were characterized to determine their biological properties in vitro and their antigenic relatedness to a previously recognized feline infectious peritonitis virus and canine coronavirus. The viruses, designated WSU 79-1146 and WSU 79-1683, were shown to have comparable growth curves with the prototype feline infectious peritonitis virus. Treatment of the feline infectious peritonitis virus strains with 0.25% trypsin indicated that they were relatively resistant to proteolytic inactivation when compared with the feline enteric coronavirus strain. This observation may serve as a useful in vitro marker to distinguish closely related members of the feline coronavirus group. Plaque assay results indicated that the feline infectious peritonitis virus strains produced large homogeneous plaques in comparison to the feline enteric coronavirus strain and canine coronavirus, which showed a heterogenous plaque size distribution. No naturally temperature sensitive mutants were detected in either of the feline coronavirus populations. Both of the viruses were antigenically related to feline infectious peritonitis virus and to a lesser extent to canine coronavirus by virus neutralization. PMID- 3038291 TI - Comparison of serological tests for the detection of antibody to natural and experimental murine cytomegalovirus. AB - Three serological tests, i.e. complement fixation test, indirect immunofluorescent assay, and enzyme-linked immunosorbent assay (ELISA) were compared for sensitivity in the detection and titration of murine cytomegalovirus antibody. The three tests were compared using sera from experimentally inoculated and naturally infected mice bled at intervals from 3 to 140 days postinfection. In the acute infection, complement fixation and indirect immunofluorescent assay tests were of comparable sensitivity for early detection of antibody, whereas the ELISA was less sensitive. In persistent infection, higher titers were recorded with ELISA. Since murine cytomegalovirus has been shown to exert significant effects on the immune response of infected mice, this antigen should be included routinely in viral antibody screening programs. PMID- 3038292 TI - Enzyme-linked immunosorbent assay for the detection of antibodies to bovine virus diarrhea virus in sera from border disease virus-infected sheep. AB - An enzyme-linked immunosorbent assay (ELISA) was established for the rapid detection of specific antibodies against the causative agent of border disease in ovine sera. Polyethylene-glycol concentrated, equilibrium density gradient purified bovine virus diarrhea virus was used as test antigen. The optimal amount of antigen was 0.5 microgram/well, and the optimal concentration of conjugate was at 1/4,000 dilution. A total of 20 ovine serum samples, which had been collected from animals with or without border disease, were compared by ELISA and serum neutralization test for the detection of border disease-specific antibodies. ELISA was shown to be equally specific but less time-consuming and easier to perform than serum neutralization test. A positive correlation (r = 0.60) between the two tests was found. PMID- 3038293 TI - The pathologic implications of vibration disease. AB - Vibration disease is a syndrome comprising a multitude of symptoms with ensuing pathologic alterations to the vasculature, nerves, and musculoskeletal system. There are various theories regarding the contributing factors involved in the etiology and pathogenesis of this disorder. This article reviews the pertinent theories and details the major aspects of vibration disease. It also introduces the potential synergistic role that the various affected systems have in influencing each other throughout the progression of this syndrome. PMID- 3038294 TI - Adenocarcinoma of the urethra in women. A clinicopathologic study. AB - Twenty-two cases of adenocarcinoma of the urethra in women were studied. Nine were classified histologically as clear cell adenocarcinoma and 13 were classified as columnar/mucinous adenocarcinoma. Thirteen patients (59%) were black. The average patient age was 61 years (range, 27 to 76 years). Follow-up ranged from 6 to 194 months, (average, 40 months). At presentation, most patients (82%) had extension of tumor into adjacent structures or metastases to regional lymph nodes. Eighty-six percent received radiation therapy and 41% underwent an anterior exenteration or cystectomy. Eight of 22 patients (36%) had no evidence of disease 21 to 194 months after diagnosis (average, 83 months). Fourteen (64%) were dead of disease 6 to 23 months after diagnosis (average, 15 months). In general, the extent of tumor correlated best with survival time. Forty-four percent of patients with clear cell adenocarcinoma were dead of disease within 24 months of diagnosis, in contrast to 77% of those with columnar/mucinous adenocarcinoma, suggesting that patients with clear cell adenocarcinoma may have a better prognosis than those with columnar/mucinous adenocarcinoma. However, the difference in survival probability between the two groups was not found to be statistically significant. Recognition of the two histologic types of urethral adenocarcinoma is important to prevent misdiagnosis of such tumors as metastases (or direct extension) of nonurethral neoplasms having a similar histologic appearance. A possible predilection of the disease for black women has not been previously described. PMID- 3038295 TI - Klinefelter's syndrome and extragonadal germ cell tumors. AB - A 25-year-old man presented with diffuse metastatic pure choriocarcinoma, thyrotoxicosis, and cardiac tamponade. No discernable testicular primary tumor was found. The patient's peripheral blood karyotype was 47, XXY and phenotypic features of Klinefelter's syndrome were present. The patient was treated with aggressive combination chemotherapy followed by salvage surgery and remains in complete remission 3 years after diagnosis. Pure choriocarcinoma, although rare as a primary testicular neoplasm, accounts for 15% of extragonadal germ cell tumors in general and 30% of germ cell tumors in patients with Klinefelter's syndrome. Historically, the diagnosis of pure choriocarcinoma has been thought to convey a very poor prognosis. The occurrence of hyperthyroidism is unique to tumors containing choriocarcinomatous elements and the management of this disorder is discussed. Treatment of extragonadal germ cell tumors is also discussed with special reference to the roles of combination chemotherapy and salvage surgery. PMID- 3038296 TI - Recurrent and metastatic pulmonary fibrous histiocytoma/plasma cell granuloma in a child. AB - A child had a pulmonary plasma cell granuloma/fibrous histiocytoma that recurred after lobectomy in 1952 and required pneumonectomy in 1953, at which time it extended into the mediastinum at the pulmonary hilus. A metastatic nodule was removed from within the diaphragmatic musculature. The long-term favorable outcome suggests a good prognosis after adequate surgical therapy. The possible relationship of fibrous histiocytoma/plasma cell granuloma to modified measles is discussed. PMID- 3038298 TI - Hepatocellular carcinoma in Chinese males and females. Possible causes for the male predominance. AB - The male-female ratio in 186 hepatocellular carcinoma (HCC) Chinese patients was 5:1. The clinical presentation, biochemical parameters, and histologic findings were the same in both sexes except for a higher proportion of underlying cirrhosis (P = 0.02), and spider naevi (P = 0.04) in the men. There were also more smokers and alcohol drinkers among the men. Over 75% of both sexes were positive for the hepatitis B surface antigen. The possible contributory factors to the predominance of males to females in HCC included: the association with the hepatitis B virus, the higher proportion of male cirrhotics, smoking, and alcohol drinking. The survival probability for both sexes was equally poor; the median survival was 8 weeks for males and 10 weeks for females. PMID- 3038297 TI - Hyponatremia in small cell lung cancer. Mechanisms not involving inappropriate ADH secretion. AB - A 62-year-old man with small cell carcinoma (oat cell type) of the lung who had hyponatremia and renal sodium loss with inappropriate antidiuresis is reported. Plasma levels of arginine vasopressin (AVP) were not elevated inappropriately. Plasma levels of atrial natriuretic peptide (ANP), however, were high, and increased after water loading and hypertonic saline infusion. The renin aldosterone axis was normal, as were adrenal, thyroid, and renal functions. Water restriction to 500 to 700 ml/d resulted in a rise in serum sodium. Analysis of the tumor tissue failed to demonstrate the presence of AVP or ANP. The findings (1) suggest that hyponatremia and renal sodium loss with inappropriate antidiuresis observed in the patient is due to an antidiuretic substance distinct from AVP, and (2) point to the possibility that hypersecretion of ANP may play a role in the pathophysiology. PMID- 3038299 TI - Serum selenium and vitamin E concentrations in families of lung cancer patients. AB - Whether or not serum selenium and vitamin E (alpha-tocopherol) concentrations were changed was examined among healthy families of lung cancer patients. Family members as a whole (115 sons and daughters of 55 patients with primary lung cancer) were found to have a trend to lower serum selenium levels (0.116 +/- SD 0.024 microgram/ml, 0.05 less than P less than 0.1). Particularly among families of adenocarcinoma patients, the mean level was significantly lower (0.111 +/- 0.019 microgram/ml, P less than 0.05) than that (0.122 +/- 0.014 microgram/ml) in age-ratio matched controls who did not have cancer patients among their second degree relatives. Serum vitamin E levels (11.85 +/- 2.85 micrograms/ml) were significantly lower among family members of adenocarcinoma patients than the controls (14.1 +/- 3.1 micrograms/ml, P less than 0.01). Serum selenium and vitamin E levels were significantly lower in lung cancer patients (n = 37, mean age, 63.9 +/- 11.2 yr) than in the controls (P less than 0.001). These data suggest that there are familial factors in serum selenium and vitamin E levels among families of lung cancer patients. PMID- 3038300 TI - Chromosome abnormalities in chronic lymphocytic leukemia revealed by cytochalasin B and Epstein-Barr virus. AB - Peripheral blood lymphocytes from eight patients with chronic lymphocytic leukemia (CLL) were cultured with Epstein Barr virus (EBV) and cytochalasin B. All eight cytochalasin B cultures had analyzable metaphases whereas only six of the EBV cultures were successful. Furthermore, the number of abnormal metaphases and the mitotic indices were greater in the cytochalasin B cultures than in the EBV cultures. Trisomy 12, alone or in combination with other abnormalities, was the most frequent cytogenetic finding. Structural abnormalities of chromosomes #6 and #14 were also found. Cytochalasin B appears to be an effective mitogen for demonstrating abnormal metaphases in patients with CLL. PMID- 3038301 TI - Patterns of ligand binding to normal, regenerating, preneoplastic, and neoplastic rat hepatocytes. AB - The binding of epidermal growth factor, asialoorosomucoid, and apoprotein E-rich lipoproteins to isolated hepatocytes was investigated at various time intervals during the step-by-step development of liver cancer in rats. The degree of binding of the three ligands showed a progressive reduction in early persistent and late persistent putative preneoplastic hepatocyte nodules. This was further decreased in hepatocytes isolated from unequivocal hepatocellular carcinomas. Regenerating liver hepatocytes bound lesser amounts of epidermal growth factor and asialoorosomucoid than did hepatocytes from control resting liver but increased amounts of apoprotein E-rich lipoproteins. The progressive decrease in ligand binding during the precancerous phase of hepatocarcinogenesis, the nodule to-cancer sequence, may render nodules less responsive to the influences of their external environments. PMID- 3038302 TI - Activation of human monocyte tumoricidal activity by C-reactive protein. AB - We examined the effect of purified human C-reactive protein (CRP) on induction of human peripheral blood monocyte (Mo)-mediated cytotoxicity (CTX) and oxidative metabolism. Exposure of Mo to acute phase serum levels of CRP in vitro resulted in dose-dependent expression of CTX against human tumor cell lines. Nonneoplastic human fibroblasts and glial cells were not affected by CRP-exposed Mo, and treatment of Mo monolayers with anti-Leu 11b (a natural killer marker) and complement did not abrogate or diminish CTX. Tumoricidal activity was observed after 20-44 h of Mo exposure to CRP, and after 48-72 h of coculture with radiolabeled target tumor cells. Mo exposed to CRP for 48 h also demonstrated elevated superoxide anion production when challenged with phorbol myristate acetate. Unlike CTX induced by lipopolysaccharide, CRP-induced CTX was completely inhibited by preincubation of CRP with phosphorylcholine, a CRP ligand, at a concentration of 5.5 molecules phosphorylcholine per molecule CRP. Further, when Mo medium (which contained 5% human AB serum) was preincubated with immobilized CRP, exposure of Mo to CRP in such medium did not result in CTX. In contrast, LPS induced CTX was not affected. CRP-induced Mo CTX was observed, however, when Mo were exposed to CRP in medium preincubated with phosphorylcholine-treated immobilized CRP, suggesting that an active serum component which complexed with CRP was not removed. These findings indicate that one of the functions of the acute phase protein, CRP, may be to active Mo and that the process may require a CRP-binding serum component. PMID- 3038304 TI - Proliferation-dependent topoisomerase II content as a determinant of antineoplastic drug action in human, mouse, and Chinese hamster ovary cells. AB - We have shown previously that quiescent Chinese hamster ovary (CHO) cells are less sensitive than log phase CHO cells to the cytotoxic and DNA cleavage effects of etoposide, a drug which appears to act via DNA topoisomerase II. This loss of sensitivity was associated with a decrease in topoisomerase enzyme activity in nuclear extracts of the quiescent cells. We have now extended our observations by examining the basis for the reduction in enzyme activity during quiescence. DNA topoisomerase II content, as assayed by immunoblotting with a polyclonal rabbit anti-topoisomerase II antiserum, was virtually absent in nuclear extracts of quiescent CHO cells in contrast to logarithmically growing cells. This suggests that the previously demonstrated loss of enzyme activity in CHO cells is a function of reduction in content rather than posttranslational modifications of the enzyme. Quiescent human lymphoblastic CCRF cells also exhibited reduced topoisomerase II content compared to actively proliferating cultures, but the difference was less than that observed in CHO cells. In contrast, log and plateau phase cultures of mouse leukemia L1210 cells exhibited similar topoisomerase II content. Reduction in enzyme content correlated with the ability of these cell lines to accumulate during quiescence with a G0-G1 content of DNA. Sensitivity to the DNA cleavage effects of etoposide in dividing and nondividing cells correlated well with enzyme content. As has been observed with CHO cells, both CCRF and L1210 cells in plateau phase were more resistant to the cytotoxic effects of etoposide than those actively dividing. The result with L1210 cells was surprising, however, in light of the equivalent DNA damage observed under the two growth conditions. Our data indicate that topoisomerase II enzyme content is proliferation dependent in some but not all cells and suggest that while enzyme content may be important in drug resistance in some cell types, other factors can decrease the sensitivity of the cell to cleavable complex formation as well. PMID- 3038303 TI - Cytotoxic response of the relatively difluoromethylornithine-resistant human lung tumor cell line NCI H157 to the polyamine analogue N1,N8-bis(ethyl)spermidine. AB - Difluoromethylornithine (DFMO), an enzyme activated irreversible inhibitor of ornithine decarboxylase (ODC), depletes intracellular putrescine, and spermidine (Spd), but not spermine, and generally leads to an inhibition of cell proliferation, without cell death, in both normal and neoplastic cells. This is the case with a culture line of human large cell undifferentiated lung cancer, NCI H157, which will survive indefinitely in DFMO containing medium and ultimately actually grows through the DFMO block. We now provide evidence that a Spd analogue, N1,N8-bis(ethyl)spermidine (BES) also suppresses ODC activity in H157 cells but leads not only to complete depletion of putrescine and Spd but also reduces intracellular spermine to 20-30% of control levels. This depletion of polyamines is accompanied by a rapid decrease in cell proliferation and ultimately cell death. The cell death resulting from BES treatment is in direct contrast to results obtained with DFMO and occurs at concentrations of less than 10 microM, whereas 5 mM DFMO is required to maintain growth inhibition in NCI H157. The observed suppression of ODC activity by BES is consistent with mechanisms by which Spd itself regulates ODC activity. Our data suggest that although both agents, DFMO and BES, interfere with polyamine metabolism, the differential sensitivities to these agents indicate susceptibility to polyamine depletion may be agent and cell type specific. Such differences may be related to the different requirement of individual cell types for polyamines and different regulatory events in polyamine biosynthesis. These differences may be exploitable in the treatment of neoplastic disease with polyamine analogues or inhibitors of polyamine biosynthesis. PMID- 3038305 TI - DNA cross-linking responses of human malignant glioma cell strains to chloroethylnitrosoureas, cisplatin, and diaziquone. AB - Cell strains derived by culture of malignant glioma (astrocytoma grade III-IV) surgical specimens were tested for the production of DNA interstrand cross-links (ISC) and DNA-protein cross-links following treatment in vitro with 1-(2 chloroethyl)-1-nitrosourea, 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU, carmustine), 1-(2-chloroethyl)-3-(2,6-dioxo-1-piperidyl)-1-nitrosourea (PCNU), cis-dichlorodiammineplatinum(II) (cisplatin), and 3,6-diaziridinyl-2,5 bis(carboethoxyamino)-1,4-benzoquinone (diaziquone). ISC and DNA-protein crosslinks were measured by means of the DNA alkaline elution technique. Large differences among the cell strains were observed in DNA cross-linking responses to individual agents. The DNA responses to the chloroethylnitrosoureas, cisplatin, and diaziquone were largely independent of each other, except for a weak correlation between ISC responses to chloroethylnitrosoureas were distributed bimodally, in accord with a phenotypic distinction between Mer+ and Mer- cells. ISC responses to cisplatin and diaziquone showed significant variation among cell strains, but the distributions were not bimodal. The results demonstrate the existence of diverse DNA cross-linking response patterns among cell strains from different tumors of a given histological type. PMID- 3038306 TI - Effect of recombinant monokines, lymphokines, and other agents on clonal proliferation of human lung cancer cell lines. AB - The modulation of clonal growth of cells of 15 human lung cancer lines was examined by coculture with different recombinant lymphokines, monokines, and several agents which induce differentiation in other malignant cell systems. Recombinant human tumor necrosis factor alpha (TNF) was inhibitory to all non small cell lung cancer cell lines with a 50% effective dose of clonal inhibition (ED50) in the range of 30-2000 units/ml. Two representative squamous lines (SK MES and P3) had 150 to 250 high affinity (Kd approximately equal to pM) cell surface TNF receptors. In contrast, clonal growth of small cell lung cancer lines was not inhibited by TNF, and two representative lines (H69c and R592) expressed negligible cell surface TNF receptors. Recombinant alpha, beta, and gamma interferons (4000 units/ml) each inhibited greater than or equal to 30% clonal growth of more than 50% of the non-small cell lung cancer lines. TNF (100-1000 units/ml) in combination with gamma-interferon was synergistic in the inhibition of clonal growth of these cells. Further studies showed that synergism of clonal inhibition occurred even when the cells were initially exposed to gamma interferon, washed, and plated in soft agar with TNF. All-trans-retinoic acid (ED50, 5 X 10(-7)-10(-6) M), dimethyl sulfoxide (ED50, 1.2-1.6%), and 12-O tetradecanoylphorbol-13-acetate (ED50, 5 X 10(-8)-10(-10) M) inhibited clonal proliferation of 7 of 9, 7 of 9, and 8 of 9 non-small cell lung cancer lines, respectively. In contrast, clonal proliferation of cells of small cell lung cancer lines was decreased only slightly at almost all concentrations of each of the agents. Interleukin-1 and -2 and granulocyte-monocyte colony-stimulating factor had no effect on the clonal growth of any of the lung cancer lines. Our results suggest that TNF in combination with gamma-interferon may be therapeutically active for some patients with non-small cell lung cancer, but small cell lung cancer probably will be unresponsive to all the agents that we examined. PMID- 3038307 TI - Mitogenic and antimitogenic transforming growth factors secreted by adenovirus 2- and simian virus 40-transformed hamster cells: possible roles in promoting tumorigenesis. AB - Adenovirus 2 (Ad2)- and simian virus 40 (SV40)-transformed hamster embryo cells differ markedly in a number of phenotypic properties including their potential for inducing tumors in hamsters. Both Ad2- and SV40-transformed cells are immortalized and readily induce tumors in immunoincompetent newborn syngeneic hamsters, but only SV40-transformed cells are highly oncogenic in both adult syngeneic and allogeneic immunocompetent hamsters. The reasons for the difference in the oncogenic potential of the Ad2- and SV40-transformed phenotypes remain elusive. However, recent studies with transforming growth factors (TGFs) indicate that these factors play an important role in determining many phenotypic characteristics of transformed cells. To determine whether TGFs secreted by Ad2- and SV40-transformed hamster embryo cells differ, we have examined the ability of media conditioned by these two transformed cell phenotypes to modulate thymidine uptake in quiescent, untransformed cells. We found that both transformed phenotypes secrete very similar TGF alpha-like mitogenic factors which inhibit binding of 125I-labeled epidermal growth factor to its receptor. Our results also show that SV40-transformed cells, but not Ad2-transformed cells, secrete a powerful mitogenic inhibitor (MI). The MI secreted by SV40-transformed cells is inhibitory for several transformed and untransformed cell types and exerts a cytostatic, not cytolytic, action on untransformed primary hamster embryo cells. MI elutes from size exclusion high-performance liquid chromatography columns with a molecular weight of 24,000. Although MI has about the same molecular weight as TGF beta, it differs from TGF beta in two important respects: it is heat labile and it has a different target specificity for antimitogenic activity. The MI secreted by SV40-transformed cells also inhibits thymidine uptake by concanavalin A-stimulated spleen lymphocytes. This finding suggests that MI might contribute to the extreme oncogenicity of SV40-transformed cells by inhibiting mobilization of immune effector cells at the site of tumor cell proliferation. PMID- 3038308 TI - Uptake and specific binding of 2,3,7,8-tetrachlorodibenzo-p-dioxin in the olfactory mucosa of mice and rats. AB - Whole-body autoradiography of mice and rats after i.v. administration of 2,3,7,8 [14C]tetrachlorodibenzo-p-dioxin ([14C]TCDD) showed a selective localization of radioactivity in the liver and nasal olfactory mucosa. In microautoradiograms of solvent extracted sections of the skulls of mice given injections of [3H]TCDD, no radioactivity was observed in the olfactory mucosa, suggesting that TCDD is not covalently bound in this tissue. The amount of specific [3H]TCDD binding sites in cytosol from the ethmoturbinates of rats (33 fmol/mg cytosolic protein) was comparable to that of the liver cytosol as estimated by electrophoresis in polyacrylamide concentration gradient gel, and therefore probably too low to explain the retention of radioactivity in the olfactory mucosa. The specific TCDD binding species in the mucosa of the ethmoturbinates exhibited a similar binding affinity for [3H]TCDD, ligand specificity, and molecular weight as the TCDD receptor from rat liver. The 7-ethoxyresorufin O-deethylase activity of the mucosa of the ethmoturbinates was induced less than twice by administration of the TCDD receptor ligand beta-naphthoflavone (5,6-benzoflavone) 40 h before killing. By administration of beta-naphthoflavone (5,6-benzoflavone) 16 h before killing, mRNA coding for cytochrome P-450d but not for cytochrome P-450c was induced to detectable levels in the mucosa of the ethmoturbinal tissue of the rat. The basal activity of 7-ethoxyresorufin O-deethylation of the mucosa of the ethmoturbinates of the rat was comparable to the corresponding activity of the liver. This basal metabolic activity of the ethmoturbinal tissue was only marginally inhibited by antibodies raised against beta-naphthoflavone (5,6 benzoflavone) induced hepatic cytochrome P-450s. Thus, enzymes other than cytochrome P-450c may possibly account for a part of the basal 7-ethoxyresorufin O-deethylase activity in the rodent olfactory mucosa. PMID- 3038309 TI - Protective influence of lactoferrin on mice infected with the polycythemia inducing strain of Friend virus complex. AB - Purified iron-saturated human lactoferrin (LF) was assessed in vivo for effects on the survival rates of C57BL X DBA/2 f1 (hereafter called BD2F1) (Fv-2sr) mice and titers of spleen focus-forming viruses (SFFV) in BD2F1 and DBA/2 (Fv-2ss) mice inoculated with the polycythemia-inducing strain of the Friend virus complex (FVC-P). LF prolonged the survival rates and decreased the titers of SFFV in mice given FVC-P. Titers of SFFV, assayed 14 days after administration of FVC-P, were measured by the spleen focus-forming unit assay in secondary mouse recipients. Decreases in titers of SFFV were apparent when LF was given in vivo as a single bolus dose of 200 micrograms within 2 h of the Friend virus complex (FVC), or as a total dosage of 200 micrograms given on days 1, 2, 4, 7, 9, and 11 after FVC-P, and to a lesser degree when LF was given as a total dosage of 200 micrograms on days 3, 4, 7, 9, and 11 after FVC-P. No decreases in titers of SFFV were detected when LF was given up to 3 days before or more than 3 days after FVC-P. LF did not appear to be directly inactivating the viruses as it did not inactivate the SFFV or the Friend murine leukemia helper virus in vitro. The results suggest that the protective effect of LF in vivo is probably due to an action on cells responding to the FVC or to an action on cells which influence the cells responding to the FVC or which influence the virus. It has been shown elsewhere that LF decreases the percentage of marrow and spleen hematopoietic progenitor cells that are in DNA synthesis in vivo and this may be the means by which the protective effect of LF is mediated in mice given the FVC. PMID- 3038310 TI - Photosensitizing effects of Photofrin II on the site-selected mitochondrial enzymes adenylate kinase and monoamine oxidase. AB - The response to Photofrin II-induced photosensitization on the activities of mitochondrial monoamine oxidase (MAO) and adenylate kinase (AK) were studied in order to gain further insight into site specific effects. Utilizing intact mitochondria in vitro, both MAO, located on the cytoplasmic side of the outer mitochondrial membrane, and AK, located in the intermembrane space, were inhibited by exposure to Photofrin II plus light; inhibition was drug-dose and light-dose dependent. However, MAO activity was inhibited to a greater extent than AK; at 35 micrograms/ml of Photofrin II and 160 J/cm2, MAO activity was decreased by 80% whereas AK activity was inhibited by 30%. Higher doses of Photofrin II had no further effect on AK activity. Studies of photosensitization of AK in different mitochondrial preparations demonstrated that inhibition of activity was evident only when mitochondrial membranes containing sequestered porphyrins were present in the reaction mixture. Using an in vivo-in vitro protocol and sampling at 2 to 72 h after administration of 25 mg/kg of Photofrin II, photosensitization of MAO (30% inhibition) was seen at 2 h after drug treatment but inhibition of activity was not observed at later times. AK activity was unchanged over the entire time course. Compared to cytochrome c oxidase, located in the inner mitochondrial membrane and which displayed a sustained inhibition of activity, we suggest that inhibition of MAO or AK activities probably does not contribute to the tumor cytotoxicity under the usual conditions used for photodynamic therapy. PMID- 3038311 TI - Induction of virus enzymes by phorbol esters and n-butyrate in Epstein-Barr virus genome-carrying Raji cells. AB - Phorbol esters and n-butyrate (SB) together could induce Epstein-Barr virus (EBV) DNA polymerase and DNase activities in Raji cells (virus nonproducer). Neither 12 O-tetradecanoylphorbol-13-acetate (TPA) nor SB alone could induce these EBV enzyme activities, transcription of the EcoRI C-region or other EBV proteins in Raji cells. The enzyme induction caused by exposure of Raji cells to TPA-SB was the result of the synthesis of virus-specified RNA, and the increase of linear EBV DNA content in Raji cells caused by TPA alone was not sufficient for induction of EBV-enzyme activities. Temporal characteristics of the TPA-SB induction process, but not the phorbol 12,13-dibutyrate-SB induction process, in Raji cells were observed; a critical phase (10-24 h) postphorbol ester treatment in phorbol 12,13-dibutyrate-SB-treated Raji cells which was responsible for the synthesis of virus RNA and enzymes was found. Phospholipase C, which increases intracellular diacylglycerols (and subsequently activates protein kinase C) was able to partially substitute for TPA in the TPA-SB induction for EBV polymerase and DNase activities. Sphingosine, a protein kinase C inhibitor, partially prevented the induction of virus enzyme activities in Raji cells treated with phorbol 12,13-dibutyrate and SB. No apparent changes in the methylation state of EBV DNA (EcoRI C region) were observed when Raji cells were treated with SB and TPA, alone or in combination. These results suggest that induction of EBV polymerase and DNase activities by TPA-SB may involve protein kinase C activation and another factor triggered by SB which together increase transcription of EBV DNA. PMID- 3038312 TI - An amplification unit in human melanoma cells showing partial homology with sequences of human papillomavirus type 9 and with nuclear antigen 1 of the Epstein-Barr virus. AB - By partial homology with the DNA of human papillomavirus type 9 a cellular amplification unit was detected which is amplified in melanoma cells but not in Epstein-Barr virus-transformed B cells of two melanoma patients. A 2.4-kilobase EcoRI fragment of this amplification unit was cloned and designated mel/HPV9. At the chromosomal level we detected mel/HPV9 in homogeneously staining regions or in abnormally banded regions containing different marker chromosomes of both melanoma cell lines. DNA sequence analysis of a part of mel/HPV 9 revealed homology with the third internal repeat array of Epstein-Barr virus nuclear antigen 1. PMID- 3038313 TI - Concurrent cisplatin and etoposide with radiotherapy in locally advanced non small cell lung cancer. AB - Twenty consecutive patients with newly diagnosed or locally recurrent non-small cell lung cancer limited to the chest were treated with concurrent radiotherapy and cisplatin plus etoposide. No patient had received prior chemotherapy or radiotherapy. Sixteen of 20 patients (80%) responded to the treatment. Median survival has not been reached but is greater than 13.5 months. Twelve patients have relapsed at a median of 10 months after starting therapy (range, 4-12 months), with only one initial relapse within the radiotherapy port. Hematologic toxicity was mild, with four of 52 wbc count nadirs less than 1000/mm3. Esophageal toxicity was also mild, with all patients maintaining oral intake throughout therapy. We conclude that simultaneous radiotherapy, cisplatin, and etoposide can be safely administered in locoregional non-small cell carcinoma of the lung. A prospective trial of this regimen versus radiotherapy alone is warranted. PMID- 3038314 TI - In vitro chemosensitivity of human lung cancer cell lines. AB - Human lung cancer cell lines, established from patients with both small cell cancer (SCLC) and non-small cell cancer (non-SCLC; squamous cell, large cell, anaplastic, and adenocarcinoma), were tested for their in vitro chemosensitivity to a panel of drugs. Drug sensitivity was assayed by either soft agar clonogenicity or a novel dye-exclusion assay. Eleven non-SCLC lines (eight continuous, three recently cultured) and five SCLC lines (all continuous) were tested. Four of eight continuous non-SCLC lines cloned sufficiently to permit limited in vitro drug testing, as did two of the five SCLC lines. All 16 cell lines could be tested for multiple drugs using the dye-exclusion assay. Drug concentrations for the nonclonogenic assay more closely approximated the area under the concentration-time curve for a given concentration of each agent. There was considerable variation in the relative sensitivity of the cell lines and the patterns of individual drug sensitivity. The majority of non-SCLC cell lines were refractory to most drugs. Cell lines derived from two previously treated SCLC patients and from three untreated SCLC patients showed greater sensitivity. Concurrent clonogenic and dye-exclusion assays showed similar drug rankings but different absolute values for percent survival. The nonclonogenic dye-exclusion assay is more rapid than the soft agar clonogenic assay (4 days vs. 2-3 weeks), could be performed on all cell lines tested, and appears to reflect the clinical diversity of human lung cancer. PMID- 3038315 TI - Phase II study of recombinant alfa-2a interferon in patients with advanced bone sarcomas. AB - Twenty previously treated patients with advanced bone sarcomas received thrice weekly im 50 X 10(6) IU/m2 doses of human alfa-interferon (interferon alfa-2a, recombinant; Roche). Seventeen patients had metastatic osteosarcomas and one each had fibrosarcoma, mesenchymal chondrosarcoma, and malignant fibrous histiocytoma. Two patients with osteosarcoma and the one with malignant fibrous histiocytoma experienced objective partial tumor regression for 1, 3, and 2 months, respectively. Fever, anorexia, myalgia, fatigue, lethargy, and moderate myelosuppression were observed commonly, and some patients developed mild nausea, vomiting, and diarrhea. No patient withdrew because of toxicity and no dose reductions were necessary except adjustments for changes in body surface area secondary to weight loss. PMID- 3038316 TI - Tamoxifen therapy in unresectable adenocarcinoma of the pancreas. AB - Twenty-four patients with biopsy-documented unresectable ductal adenocarcinoma of the pancreas were treated with tamoxifen. Six of 15 postmenopausal female patients had documented prolonged survival, with three living greater than 2 years. PMID- 3038317 TI - Phase II treatment of central nervous system gliomas with high-dose etoposide and autologous bone marrow transplantation. AB - A total of 16 patients with progressive CNS gliomas who had previously received maximal radiation therapy and chemotherapy were treated on a phase II study using high-dose etoposide with autologous bone marrow transplantation to assure hematopoietic recovery. Three patients (19%) responded with both improved neurological status and decreased mass on computerized tomographic scan. Median survival for all treated patients was 4 months with the three responders living 9, 10, and 54+ months. PMID- 3038318 TI - Phase II trial of oral piritrexim (BW301U) in patients with stage III non-small cell lung cancer. PMID- 3038319 TI - Phase II trial of esorubicin in patients with advanced non-small cell lung cancer. PMID- 3038320 TI - Transient tumor marker surge following chemotherapy of testis tumors. PMID- 3038321 TI - [Mitochondrial function and formation of superoxide radicals in the hypertrophic cardiac muscle: effect of the administration of coenzyme Q10]. PMID- 3038322 TI - [Regulation of hormonal sensitivity of cardiac cells in culture by polyamines]. PMID- 3038324 TI - Role of neutrophils in ischemic heart disease: pathophysiologic role in myocardial ischemia and coronary artery reperfusion. PMID- 3038323 TI - Leukotrienes: role in cardiovascular physiology. AB - Our current understanding of the physiology of the leukotrienes is far from complete. The abundant supply of synthetic products has directed researchers into examining what the mediators affect rather than the basic mechanism studies of their involvement in disease. It is clear that the peptide leukotrienes possess potent constrictor actions of the microvasculature and can enhance permeability. These actions alone represent a new avenue of interpreting pathologic processes and could lead to alternate means of treating certain diseases in the future. It is of special interest that a consistent action of the leukotrienes is to reduce coronary blood flow, decrease myocardial contractility, and reduce cardiac output without affecting the heart rate. This profile of action is the first indication that a mediator can play a significant role in unstable angina. The main physiologic actions of the leukotrienes in the cardiovascular system are currently believed to be associated with episodes of ischemia and shock. Their relative contribution to the shock states, especially when compared with the actions of other known mediators of shock such as the prostaglandins, thromboxane, angiotensin, serotonin, and histamine, awaits clarification. LTB4 is a proinflammatory mediator that has opened a completely new perspective on the physiologic role of phagocytic cells. Novel therapeutic approaches to inflammatory-related diseases may result from an inhibition of cell chemokinesis, aggregation, and degranulation. The role of LTB4 in the immune system awaits further clarification. PMID- 3038325 TI - Physiologic and pathophysiologic role of cyclo-oxygenase metabolites of arachidonic acid in circulatory disease states. AB - The overall assessment of eicosanoids in ischemia and shock is complex. Certain prostaglandins, notably PGI2, actually improved survival in shock and ischemic states; however, some, including TxA2, act as mediators, significantly contributing to the pathophysiology of vasospasm and ischemia. This makes the intelligent use of eicosanoid-related drugs very difficult. Broad-based inhibitors (for example, cyclo-oxygenase inhibitors) are not likely to exert significant protective effects because they inhibit beneficial as well as detrimental eicosanoids. More appropriately, the therapy of ischemic disorders should be designed to employ specific agents (for example, TxSI and TxRA) rather than broad-based agents. Preliminary studies already suggest the success of such an approach. Future investigations will focus on the application of potent synthetic modulators or analogues of specific eicosanoids in an attempt to prevent the deleterious effects of eicosanoid mediators of ischemic disorders and to potentiate and prolong the beneficial actions of therapeutic eicosanoids. PMID- 3038326 TI - Effects of enalapril maleate on blood pressure, renin-angiotensin-aldosterone system, and peripheral sympathetic activity in essential hypertension. AB - Recent experimental studies showed that inhibition of angiotensin II synthesis may reduce sympathetic activity as evaluated by plasma catecholamine assay, sharing in the antihypertensive effect of angiotensin converting enzyme (ACE) inhibitors. Fifteen patients with essential hypertension were studied. Blood pressure and heart rate were evaluated both at rest and after stressor laboratory tests, before and four hours after administration of 20 mg of enalapril maleate and on the 14th and 120th days of continued administration. At the same time, blood samples were drawn for determinations of plasma renin activity, ACE, angiotensin II, plasma aldosterone concentration, and plasma norepinephrine levels. Enalapril in a dosage of 20 mg/day significantly and progressively lowered systolic and diastolic blood pressure at rest, with maximal decreases observed on the 120th day of the study period (P less than 0.001). Heart rate at rest and after exercise showed no significant differences throughout the study period. Good blood pressure control was observed during stressor laboratory tests. The greatest impact of blood pressure was observed on the 120th day during dynamic exercise (mean blood pressure from 139 +/- 3.9 to 111.5 +/- 6.3 mmHg; P less than 0.01) and on the 14th day during the cold pressure test (mean blood pressure from 133.3 +/- 3.9 to 111.2 +/- 4.7 mmHg; P less than 0.005). A marked and persistent ACE inhibition and a gradual and progressive decrease of angiotensin II (from 12.42 +/- 2.15 to 5.45 +/- 1.68 pg/ml; P less than 0.005) characterized the humoral activity of enalapril maleate. Moreover, a significant decrease of plasma norepinephrine levels was observed during the follow-up period with maximal reduction on the 120th day (from 311 +/- 34 to 197 +/- 33 pg/ml; P less than 0.01). It has been demonstrated that the pressor effect of angiotensin II was blunted during exercise. Our hemodynamic and humoral results appear to confirm the hypothesis that enalapril maleate may reduce blood pressure by direct inhibition of ACE and of kininase II as well as by a decreased sympathetic output, which may be secondary to angiotensin II inhibition. These results agree with the recent experimental demonstration of a reduced sympathetic nervous response to nerve stimulation during ACE inhibition. PMID- 3038328 TI - Implications of disaggregation procedures on biological representation of human solid tumours. AB - This study was designed to define some biological aspects of cell suspensions, obtained by mechanical or enzymatic disaggregations, and to verify whether single cell suspensions are representative of original solid tumours. The study was performed on a series of 25 human solid tumours including breast carcinoma, ovarian carcinoma and malignant melanoma. A higher cell viability and a loss of aneuploid subpopulations, or a lower fraction of aneuploid cells, were observed in enzymatically-released samples than in samples obtained by the mechanical procedure. Moreover, the proliferative activity, which was generally similar for the cell suspensions obtained by the two disaggregation procedures, was always markedly lower in the cell suspensions than in solid samples from the same tumour. In conclusion, the results from this study indicate that many changes, such as selective release of cell populations from the tumour matrix, damage and destruction of aneuploid and proliferating cells can be induced to various extents by different disaggregation procedures. PMID- 3038327 TI - Purine catabolism in cafeteria-diet induced obesity in the rat. PMID- 3038329 TI - Effects of a prolonged treatment with aminoglutethimide on the zona fasciculata of rat adrenal cortex: a morphometric investigation. AB - The effects of a 7-day administration of aminoglutethimide (AG) on the adrenal zona fasciculata were examined in "normal" and dexamethasone/ACTH-treated rats. There was a 70-74% decrease in the concentration of corticosterone in blood, but no conspicuous qualitative changes suggesting cell degeneration occurred. Morphometry showed that AG induced a significant hypertrophy of the zona fasciculata and its parenchymal cells only in "normal" animals, which was due to an increase in the volume of the mitochondrial compartment and to proliferation of the smooth endoplasmic reticulum. This response to AG was considered to be non specific and mediated by the enhanced secretion of ACTH following the decrease in the blood level of corticosterone. AG administration significantly increased the volume of the lipid-droplet compartment and the number of intramitochondrial lipid-like inclusions in both groups of animals. These changes were interpreted as the morphological counterpart of the AG-induced block of cholesterol utilization in steroid synthesis. PMID- 3038330 TI - Binding and internalization of native gonadoliberin (GnRH) by anterior pituitary gonadotrophs of the rat. A quantitative autoradiographic study after cryoultramicrotomy. AB - To identify anterior pituitary cell types containing GnRH binding sites and to study the internalization process of this peptide by target cells under physiological conditions, autoradiography was performed on rat anterior pituitaries removed at specific time intervals (2-60 min) after intravenous injection of mono-radioiodinated 125I-GnRH into intact males. At electron microscopic level, gonadotrophs and lactotrophs appeared to contain silver grains. Concomitant administration of an excess of unlabeled GnRH with the radioiodinated hormone prevented this localization indicating the specificity of the reaction. The time-course study in gonadotrophs showed that 2 min after injection silver grains could be found over the plasma membrane, secretory granules and nuclear membrane. Similar results were observed 5 and 15 min after injection. Extensive label was observed over the nucleus and nuclear membrane 15 to 60 min after injection. The injection of a radioiodinated GnRH agonist [D Trp6, Pro9 (Net), DesGly10]-GnRH produced comparable results. In contrast, the injection of 125I-[D-pGlu1, D-Phe2, Trp3,6]-GnRH, an antagonist of GnRH, produced positive labeling only at the plasma membrane without internalization. These results indicate that, after binding with receptors on the plasma membrane, GnRH is rapidly internalized, accumulating in secretory granules, and localizing over the nuclear membrane and later, in the nucleus. Association of radioactivity with secretory granules could be related to a specific action of GnRH at this level or to receptor recycling, and presence of label in the nucleus may be related to stimulation of neosynthesis of LH and GnRH receptors. PMID- 3038331 TI - Activation of calmodulin by the essential trace element chromium. AB - Chromium at very low concentrations is an essential trace element--at higher concentrations it is associated with contact dermatitis and other toxicity problems. Its ionic radius is just outside that of other metal cations which have been found to activate calmodulin in vitro. We found that chromium was able to activate calmodulin at two different concentration ranges--over the micromolar range (which would probably never be achieved in man) a small degree of activation was found--but a much greater activation (76% of the maximum possible) was also found at nanomolar concentrations of chromium. In welders, who work with stainless steel and who were not reporting any physical symptoms of chromium toxicity, red cell chromium levels were 28.2 +/- 3.3 nM (n = 22) compared to 7.5 +/- 0.7 nM (n = 11) for normal controls. Thus, the concentration of chromium experienced within the cell can be of the order which will activate calmodulin in vitro. The possibility exists, therefore, that inappropriate activation of calmodulin could be relevant to chromium biology possibly contributing to the symptoms of chromium toxicity. PMID- 3038332 TI - An inducible mammalian amber suppressor: propagation of a poliovirus mutant. AB - We describe a general protocol for controlled gene amplification, which allows conditional expression of high levels of amber suppressor activity in monkey kidney cells, and we demonstrate its use in the genetic analysis of animal viruses by the generation and propagation of the first nonsense mutant of poliovirus. A human amber suppressor tRNASer gene linked to the SV40 origin of replication and a second DNA carrying a temperature-sensitive SV40 large T antigen gene were cotransfected into monkey cells. Cell lines having stably integrated the DNAs were isolated. Shifting the cells from the nonpermissive temperature to a lower permissive temperature caused the amplification of the suppressor tRNA gene, which resulted in suppression efficiencies at amber codons of 50%-70%, as measured by suppression of an amber codon in the E. coli chloramphenicol acetyltransferase gene. A mutant of poliovirus, in which a serine codon in the replicase gene was converted to an amber codon, was efficiently propagated on the suppressor-positive cell lines. The mutant virus reverted to wild-type by a single base change to a serine codon at a frequency of approximately 2.5 x 10(-6), surprisingly low for a RNA genome. PMID- 3038333 TI - The effect of protein context on nuclear location signal function. AB - The effect of position and number and of another intracellular location signal on the activity of the nuclear location signal was investigated. A minimal signal was inserted into several sites within the polypeptide chain of pyruvate kinase. The results observed suggest that a nuclear location signal can function at a variety of positions within a protein but that in some locations its activity is masked. Multiple copies of a partially defective signal were integrated into pyruvate kinase. The data suggest that multiple signals can cooperate to enhance nuclear accumulation. A nuclear location signal failed to function when inserted into polyomavirus middle T but was active in an identical variant lacking the carboxy-terminal hydrophobic tail. We conclude that while a minimal nuclear location signal is sufficient for nuclear localization, its activity is crucially dependent on the protein context within which it is present. PMID- 3038334 TI - The physical map of the whole E. coli chromosome: application of a new strategy for rapid analysis and sorting of a large genomic library. AB - Thirty-four hundred lambda phage clones containing segments of the E. coli chromosome were isolated and used to construct a 4700 kb long integrated restriction map for eight six-base-recognizing enzymes by a rapid mass-analysis method. Our strategy was to measure the sizes of partial restriction enzyme digests by hybridization with a vector probe in a manner analogous to nucleotide sequencing. The data were sorted into groups by a computer program and the boundary clones were further correlated with each other using a mass hybridization method. These clones can be exploited for the isolation of any desired E. coli genes if their map positions are known. Also, the strategy is applicable to analyses of the genomes of other organisms. PMID- 3038335 TI - Semi-intact cells permeable to macromolecules: use in reconstitution of protein transport from the endoplasmic reticulum to the Golgi complex. AB - We introduce a new method that removes portions of the plasma membrane of eukaryotic cells to form semi-intact cells. During preparation, these cells lose their soluble cytoplasmic contents, but retain secretory organelles such as the ER and Golgi complex in an intact form. Transport of protein between the ER and Golgi can be functionally reconstituted in vitro using these semi-intact cells by incubation in the presence of cytosol and ATP. Export of the vesicular stomatitis virus strain tsO45 G protein from the ER in vitro is temperature-sensitive, similar to the result observed in vivo. These cells allow direct access of chemicals and antibodies to the cytoplasmic domain of the cell and may be a widely applicable model system for study of a broad range of problems in cell biology. PMID- 3038336 TI - Reactivation of spindle elongation in vitro is correlated with the phosphorylation of a 205 kd spindle-associated protein. AB - Mitotic spindles isolated from the diatom Stephanopyxis turris consist of two half-spindles of closely interdigitating microtubules that slide relative to one another in the presence of ATP, reinitiating spindle elongation (anaphase B) in vitro. Purified spindles that have been exposed to ATP-gamma-S undergo ATP dependent reactivation more readily than do control spindles. Thiophosphorylated proteins in such spindles are located in the spindle midzone, kinetochores, and a portion of the pole complex. One major thiophosphorylated peptide of 205 kd is detected in extracts prepared from spindles labeled with [35S]ATP-gamma-S, and is also localized in the spindle midzone by using an antibody that recognizes thiophosphorylated proteins. It is likely that this 205 kd peptide is either a positive regulator or mechanochemical transducer of microtubule sliding when it is in a phosphorylated state. PMID- 3038337 TI - Enhancement of mRNA nuclear transport by promoter elements. AB - This report describes studies on the kinetics of herpes thymidine kinase (TK) mRNA transport in X. laevis oocytes as studied by microinjection and microdissection. We show that TK mRNA nuclear transport in this cell can be dramatically altered by introduction of specific DNA sequences into the nucleus. Introduction of DNA sequences encompassing the promoter results in enhancement, in trans, of a transport process that can deliver previously synthesized RNA to the cytoplasm. The report suggests that the promoter of a eukaryotic gene can functionally interact with the mechanism involved in mRNA nuclear transport. PMID- 3038338 TI - Rat macrophage treatment with lipopolysaccharide leads to a reduction in respiratory burst product secretion and a decrease in NADPH oxidase affinity. AB - The effect of LPS on the respiratory burst in resident rat peritoneal macrophages has been examined. Rat macrophages secreted high levels of both O2- and H2O2 in response to triggering with phorbol esters, opsonized zymosan, and immune complexes. After culture in vitro with LPS these macrophages exhibited a marked diminution in their capacity to secrete high levels of respiratory burst products. The LPS-mediated loss of secretory activity was apparent after 2 hr of exposure to LPS and was inhibitable by polymyxin B in a dose-dependent fashion. The effect was not selective for any triggering agent type as inhibition of secretory activity occurred after triggering with PMA, zymosan and immune complexes. PGE2 added at levels secreted by the macrophages in response to LPS also inhibited respiratory burst product secretion. In addition, indomethacin prevented the LPS-mediated inhibition of secretion. Because the inhibition of secretion was common to all triggering agents tested, this suggested that the basis for the impaired secretion was at a level other than the receptor for the triggering agent. Both LPS and PGE2 treatment of the macrophages increased the Km of the oxidase for NADPH (1.7- to 2.3-fold) without affecting significantly the Vmax of the enzyme. These data suggest that stimulation of rat peritoneal macrophages by LPS results in an impaired ability to secrete respiratory burst products as a result of a PGE2-mediated decrease in NADPH oxidase affinity and that this alteration is independent of alterations in tumoricidal activity. PMID- 3038339 TI - Differences of T-cell activation by the anti-CD3 antibodies Leu4 and BMA030. AB - Two anti-CD3 antibodies and their Fab/F(ab')2 fragments were compared with regard to their requirement for secondary signals and generations of intracellular messengers. The anti-CD3 antibody BMA030 was found to require monocyte contact to elicit T-cell mitogenesis. Cross-linking by plastic-bound goat anti-mouse antibodies (panning) failed to activate T cells, even in the presence of recombinant IL-1 or IL-2. In contrast, crosslinking of the anti-CD3 antibody Leu4 or Leu4 fragments was mitogenic in monocyte-free cultures. Measurements of intracellular Ca2+ ([Ca2+]i) and generation of inositol phosphates revealed that binding (+/- panning) of BMA030, Leu4, and their F(ab')2 fragments generated similar amounts of intracellular messengers and thus failed to explain the different responsiveness to passive crosslinking. Since the generation of these messengers was not necessarily followed by proliferation but was always observed when mitogenesis occurred, we conclude that the elevation of [Ca2+]i and the production of inositol phosphates are required but not sufficient to trigger mitogenesis. PMID- 3038340 TI - Biochemical basis of HLA-DR and CR3 modulation on human peripheral blood monocytes by lipopolysaccharide. AB - The biochemical events leading to enhanced membrane expression of HLA-DR and CR3 by human peripheral blood monocytes (MO) following exposure to bacterial lipopolysaccharide (LPS) were examined. In a previous study we demonstrated that an increase in intracellular calcium was necessary, but not sufficient, for MO to increase membrane expression of both antigens within 1 hr of addition of LPS. The present study was initiated to examine the other biochemical requirements which lead to the MO response to LPS. Enhanced expression of both antigens following addition of LPS was dependent on microfilament function, but independent of microtubule function and of protein synthesis. Inhibition of formation of cyclooxygenase or lipoxygenase metabolites of arachidonic acid had no effect on HLA-DR or CR3 modulation by LPS. A role for phosphatidylinositol metabolism was suggested by the inhibition of the MO response to LPS by dibutyryl cAMP and theophylline and by the enhanced expression of both antigens following addition of phorbol diesters. However, H-7, a putative inhibitor of protein kinase C, did not alter the MO response to LPS or phorbol diesters. These results suggest that LPS enhances expression of HLA-DR and CR3 by inducing redistribution of these antigens from an intracellular pool. The data also support a role for the generation of hydrolysis products of phosphatidylinositol, leading to calcium redistribution and activation of protein kinase C or other kinases, in the MO response to LPS. PMID- 3038341 TI - Epstein-Barr virus-transformed lymphoblastoid cell lines as antigen-presenting cells and "augmenting" cells for human CMV-specific Th clones. AB - The ability of Epstein-Barr virus-transformed lymphoblastoid cell lines (LCL) to present human cytomegalovirus (HCMV) antigen to a panel of HCMV-specific T helper (Th) clones was evaluated. Among the seven Th clones studied, only one clone (SP CN/T3-16) proliferated well to HCMV presented by both autologous mononuclear cells (MNC) and LCL, and one clone (SP-CN/T3-9) proliferated significantly better to HCMV presented by autologous LCL than by autologous MNC. The majority of the HCMV-specific Th clones tested (five out of seven) responded much better to HCMV presented by MNC than to HCMV presented by LCL. The mechanism(s) responsible for the inefficiency of LCL to present HCMV to certain clones was studied. Our results suggested that the defect of LCL is not due to insufficient interleukin 1 production, insufficient MHC class II molecule expression, nor an inhibitory mechanism or factor. In this report, we also demonstrate that by adding a minimum amount of LCL along with MNC as antigen-presenting cells (APC), one can restimulate and expand Th clones much more efficiently than by using MNC alone as APC. PMID- 3038342 TI - Sodium-dependent and sodium-independent phosphate uptake by full-grown, prophase arrested oocytes of Xenopus laevis before and after progesterone-induced maturation. AB - The uptake of inorganic phosphate by full-grown, prophase-arrested oocytes occurs by two essentially independent transport systems, which accomplish the delivery of extracellular phosphate into two different cellular compartments. One transport system is sodium-dependent, the other is sodium-independent. Both systems can be inhibited by sulfhydryl reagents and the sodium-dependent system becomes inhibited during progesterone-induced oocyte maturation. PMID- 3038343 TI - Potentiation of nerve growth factor(NGF)-mediated neurite outgrowth in high K+ medium is associated with increased binding of iodinated NGF in PC12 cells. AB - Nerve growth factor(NGF)-mediated neurite outgrowth of PC12 pheochromocytoma cells was potentiated in medium containing high concentrations of extracellular K+. The binding of iodinated NGF to the cells was also enhanced by raising the concentration of K+ in medium up to 100 mM; the enhancement was saturated at 50 mM K+. Although the mechanism by which NGF-mediated neurite outgrowth is potentiated in high K+ medium remains to be largely unknown, high K+-induced alterations in the NGF binding are suggested to play a role in this phenomenon. PMID- 3038344 TI - Studies on griseolic acid derivatives. IV. Synthesis and phosphodiesterase inhibitory activity of acylated derivatives of griseolic acid. PMID- 3038346 TI - [Pleomorphic xanthoastrocytoma versus glioblastoma multiforme]. PMID- 3038345 TI - Studies on peptides. CXLVIII. Application of a new deprotecting procedure with trimethylsilyl trifluoromethanesulfonate for the syntheses of two porcine spinal cord peptides, neuromedin U-8 and neuromedin U-25. PMID- 3038347 TI - [Histopathological classification of breast cancer: a clinicopathologic analysis of 245 cases]. PMID- 3038348 TI - [Clinico-pathological analysis and ultrastructural study of 141 cases of stomach carcinoma]. PMID- 3038349 TI - [Electron microscopic study of human T-cell leukemia virus]. PMID- 3038350 TI - [Studies on the inhibitory effects of several TDA analogs against herpes simplex virus DNA synthesis]. PMID- 3038351 TI - [Preliminary study on the fertilization of Ligusticum chuanxiong]. PMID- 3038352 TI - [Treatment of malignant trophoblastic neoplasia with trichosanthin]. PMID- 3038353 TI - [Epidemiology of Pseudomonas aeruginosa hospital infection in the Ivory Coast]. AB - 210 P. aeruginosa hospital strains isolated at Abidjan from pathological samples or contaminated material were studied by means of serotyping, biotyping, phage typing and for their resistance to eleven antibiotics. 87% were classified into 14 serogroups: O:1, O:2, O:3, O:4, O:5, O:6, O:8, O:9, O:10, O:11, O:12, O:13, O:15, O:16. No strains were O:7 or O:14. The presence of an ortho-nitrophenyl beta-d-galactopyranoside hydrolase was demonstrated in 42 strains belonging to the serogroup O:11. Sixty-five phage-types were defined but 37% of strains were untypable by phages. The incidence of resistance to each tested antibiotic was as follows: carbenicillin 49%, mezlocillin 44%, ticarcillin 34%, azlocillin 25%, cefamandole 94%, latamoxef 15%, cefotaxime 41%, cefsulodin 32%, tobramycin 35%, amikacin 7%, gentamicin 39%. Thirty-eight strains were multiresistant to carbenicillin, mezlocillin, ticarcillin, azlocillin, and four strains to eleven antibiotics. These results are compared with data of the scientific literature on the epidemiology of hospital Pseudomonas infection. PMID- 3038354 TI - [Hemorrhagic fever viruses. Principal epidemiologic aspects]. PMID- 3038355 TI - [Human arbovirus infections in Burundi: results of a seroepidemiologic survey, 1980-1982]. AB - A serological survey on 623 human sera was conducted in Burundi in 1980-1982, in order to evaluate the frequency of arboviral antibodies in the inhabitants of the three main areas: lowlands, central plateau and mountainous ridge. The results show a rather high activity of arboviruses, mainly in the lowlands (34.2% of inhabitants with antibodies). Chikungunya virus seems to be the most active arbovirus; the activity of Flavivirus is moderate; no trace of activity of yellow fever or West Nile viruses was found; Bunyavirus antibodies (particularly against Ilesha virus) were also detected. PMID- 3038356 TI - [Surveillance of enteroviruses in the waste water of the Ivory Coast]. AB - Viral water contamination is on the increase all over the world and one is concerned by the dangers to which it exposes health and preoccupied with the search for effective solutions. This study, which was conducted in a year in Cote d'Ivoire, enabled us to examine the degree of viral pollution from waste water collected from a lagune. It allowed us to follow the distribution of enteroviruses and their fluctuations during the year. 164 waste water samples were tested and 93 enterovirus strains isolated. 67.89% showed polioviruses and 32.11% coxsackieviruses. Seasonal fluctuations of the virus concentration in the lagune water were shown from statistical analysis of the data collected. Concentration was high during the rainy season and definitely lower during the dry season with the rise in water temperature. PMID- 3038357 TI - Molecular epidemiology of influenza viruses: memorandum from a WHO meeting. AB - This Memorandum summarizes recent information on influenza viruses of non-human hosts, discussed at the WHO Consultation on Molecular Epidemiology of Influenza Viruses, Athens, Georgia, USA, in September 1986. It was noted that a wide variety of antigenic variants of influenza viruses have been isolated from non human hosts, especially birds. Of particular epidemiological interest in recent years has been the isolation of H7N7 influenza viruses from epidemics of disease in seals, the isolation of an H10N7 virus from outbreaks of disease among domestic mink, and the occurrence of pathogenic avian influenza viruses in Australia, Ireland and the USA. PMID- 3038358 TI - [Frequency of nosocomial measles in a maternal and child health center in Abidjan]. PMID- 3038359 TI - Enhanced expression of insulin-like growth factor II is not a necessary event in Wilms' tumour progression. AB - Wilms' tumour (WT) is a paediatric kidney tumour arising from the embryonal metanephrogenic blastema. Recent reports suggest that the expression of messenger RNA (mRNA) for insulin-like growth factor II (IGFII) is elevated in WT. Total cytoplasmic RNA was extracted from 11 sporadic WTs and analysed for IGFII mRNA using dot-blot hybridization. The level of IGFII mRNA expression varied greatly and not all tumours displayed enhanced IGFII expression. Two successive WT xenografts were established in nude mice. The original WT and first passage xenograft showed a blastematous histology, while the second passage xenograft showed epithelioid differentiation and tubule formation. Analysis of the expression patterns of these xenografts showed elevated IGFII expression in the primary undifferentiated tumour and the second differentiated xenograft, while the first undifferentiated xenograft failed to exhibit enhanced IGFII expression. These data show that elevated IGFII mRNA is not an essential component of the progression of WT and that WT tumourigenicity is independent of the level of IGFII expression. Therefore, IGFII overexpression in WT is most likely a tumour epiphenomenon. PMID- 3038360 TI - Gene expression rather than the initiation of DNA replication is the principal target of lethal u.v.-induced damage in a regulatory region of SV40 DNA. AB - The survival of transfected simian virus (SV) 40 DNA is acutely sensitive to damage in a 302-bp regulatory region that governs viral gene expression and the initiation of viral DNA replication. We investigated whether the lethal effect of damage in this region is due to the disruption of gene expression or to the inhibition of DNA replication by comparing the survival of damaged viral DNA in CV-1 and cos-1 African green monkey cells. Viral early sequences integrated into the genomic DNA of cos-1 cells complement the growth of virus with defective early genes and were therefore expected to reverse viral sensitivity to lesions that interfere with early gene expression. Our results indicate that viral sensitivity to damage in the regulatory region is almost completely abolished in cos-1 cells. This finding identifies gene expression rather than the initiation of DNA replication as the major target for lethal damage in that portion of the SV40 genome. Sensitivity to damage in the viral late gene region is the same in CV-1 and cos-1 cells, indicating that cos-1 cells are not merely more proficient in host-cell reactivation. Our results allow us to partition the overall lethal effect of DNA damage into sectors, and to assign each sector to the disruption of a particular genetic function. PMID- 3038361 TI - N-methyl-N'-nitro-N-nitrosoguanidine and benzo[a]pyrene-7,8-diol-9,10-epoxide inhibit glucocorticoid-inducible polyoma virus middle-T gene expression in rat mT 1 cells by a post-transcriptional mechanism. AB - Treatment of rat mT-1 cells with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and benzo[a]pyrene-7,8-diol-9,10-epoxide (BPDE) inhibited the steroid-induced expression of the polyoma virus middle-T (mT) gene. Rat mT-1 cells were derived from normal F111 fibroblasts transfected with a plasmid (pSVM-mT) containing a mT cDNA sequence linked to the mouse mammary tumor virus long terminal repeat, thus rendering the mT gene responsive to glucocorticoid hormones. Following a 1-h treatment period with either 100 ng/ml of MNNG or 100 ng/ml of BPDE, administration of 1 microM dexamethasone (DEX) for 24 h resulted in, respectively, 51 and 33% less induction of phosphorylated mT protein in carcinogen-treated than in solvent-treated cells. MNNG appeared to increase the basal level of phosphorylated mT antigen expression by 44%, whereas BPDE had little or no effect on the level of phosphorylated mT protein in uninduced cells. MNNG and BPDE had little or no effect on the level of total steroid-induced mT RNA following addition of DEX for 4 or 20 h. The concentrations of MNNG and BPDE used in these experiments caused, respectively, an 18 and 30% decline in cell number 24 h following carcinogen treatment and had no effect on cell viability as determined by trypan blue exclusion. Previous studies demonstrating the inhibitory effects of chemical carcinogens on the steroid-mediated induction of endogenous genes suggested that carcinogens mediate their effects via a pretranslational mechanism. The present study, using a recombinant DNA construct inserted into the cell genome, provides evidence that carcinogens can inhibit hormone-induced gene expression by a post-transcriptional mechanism as well. PMID- 3038362 TI - Effect of cytochalasin B on glucose uptake, utilization, oxidation and insulinotropic action in tumoral insulin-producing cells. AB - Cytochalasin B (17-3 microM) virtually abolished 3-O-methyl-D-[U-14C]glucose uptake and D-[5-3H]glucose utilization in tumoral insulin-producing cells of the RINm5F line. This coincided with a marked decrease in D-[U-14C]glucose oxidation and suppression of the stimulant action of D-glucose upon insulin release. Cytochalasin B, however, augmented basal insulin release by the tumoral cells. The RINm5F cells appeared much more sensitive than normal islet cells to cytochalasin B, as judged by the relative magnitude of inhibition in either hexose uptake or utilization. In both cell types, the inhibitory action of cytochalasin B upon glucose metabolism seemed to be competitive, being more marked at low than high glucose concentration. These results are interpreted in support of the view that a decreased efficiency of hexose transport across the plasma membrane represents an essential deficiency of the RINm5F cells. PMID- 3038363 TI - Prolyl hydroxylase activity as an index of liver damage induced by ethanol. AB - It was found that chronic intoxication of rats with ethanol results in an increase of prolyl hydroxylase activity in liver and serum of the experimental animals. The increase of enzyme activity precedes the morphological symptoms of liver damage. The possibility arises that the assay of prolyl hydroxylase in serum or in liver biopsy samples could be useful for the diagnosis of the tendency of some individuals to develop liver cirrhosis induced by ethanol. PMID- 3038364 TI - Beneficial effects of a leukotriene antagonist on endotoxin-induced acute hemodynamic alterations. AB - The role of lipoxygenase metabolites of arachidonic acid as inflammatory mediators of endotoxin shock remains uncertain. In this study the effect of LY171883, a selective leukotriene D4/E4 antagonist, on the hemodynamic alterations of endotoxin shock was assessed. Adult male rats were given an intraperitoneal injection of LY171883 (30 mg/kg) or vehicle 10 minutes prior to intravenous injection of endotoxin (15 mg/kg) or vehicle. Cardiac output, mean arterial pressure, and multiple organ blood flows were determined at 30 minutes after endotoxin or vehicle administration with 85Sr-radiolabeled microspheres. Cardiac output decreased from 32.1 +/- 2.7 ml/min/100 g in the control group to 16.3 +/- 2.7 ml/min/100 g in endotoxin-treated animals (P less than .05). Pretreatment with LY171883 blunted significantly (P less than .05) this fall in cardiac output (26.3 +/- 2.6 ml/min/100 g). Endotoxin reduced mean arterial pressure from 95 +/- 8 mm Hg in controls to 57 +/- 8 (P less than .05), which was not, however, different from control values in rats receiving the LTD4/E4 antagonist. There was also significant (P less than .05) blunting of the endotoxin-induced fall in blood flow to the heart, gastrointestinal tract, and kidneys in animals pretreated with LY171883. Our data demonstrate that this selective leukotriene D4/E4 antagonist has significant salutary actions in endotoxin shock and suggest that LTD4 and/or E4 mediate, in part, the acute hemodynamic sequelae of endotoxin shock. PMID- 3038365 TI - Effects of atrial natriuretic factor on the vasoconstrictor actions of the renin angiotensin system in conscious rats. AB - Previous studies have indicated that the hypotensive effects of atrial natriuretic factor were enhanced in renin-dependent hypertensive rats, suggesting that the atrial peptides may antagonize the vasoconstrictor effects of the renin angiotensin system. The present study was designed to define further the interaction between atrial natriuretic factor and the renin-angiotensin system by examining the hemodynamic effects of Wy-47,663, a synthetic human atrial natriuretic factor, in conscious normotensive rats, in renin-dependent (aortic ligated) hypertensive rats, and in rats made hypertensive by chronic infusion of angiotensin II. Changes in renal and mesenteric blood flow were continuously monitored in the rats using pulsed Doppler flow probes chronically implanted in the animals one week prior to testing. Infusion of increasing doses of Wy-47,663 caused dose-dependent reductions in mean arterial pressure in all three groups of rats, but the depressor responses were significantly greater in renal hypertensive and angiotensin II-infused rats. Renal blood flow tended to increase during the infusion of the atrial peptide in the angiotensin II-treated rats, and renal vascular resistance fell significantly (-37 +/- 6%). However, Wy-47,663 significantly reduced renal blood flow in the normotensive and renal hypertensive rats, while renal vascular resistance was increased (29 +/- 6%) and unchanged (3 +/- 9%), respectively. Mesenteric blood flow was reduced significantly, and mesenteric vascular resistance was increased markedly in all three groups of rats during infusion of the atrial peptide. In a separate group of renal hypertensive rats, the hemodynamic effects of complete blockade of the renin-angiotensin system were assessed by injection of an angiotensin II converting enzyme inhibitor (Wy-44,655).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3038366 TI - Granulocytes cause reperfusion ventricular dysfunction after 15-minute ischemia in the dog. AB - Regional ventricular dysfunction (the stunned myocardium) persists for several hours after 15 minutes of ischemia and reperfusion in the dog. Superoxide-radical induced damage appears to be one of the mechanisms of this injury. We tested whether granulocytes were a direct source of injury in the stunned myocardium in the 15-minute ischemia dog model. Regional function during agranulocytic extracorporeal coronary perfusion (using Leukopak filters) with ischemia and reperfusion was compared with function during a second period of ischemia and reperfusion after removal of the filters (granulocytopenia). Flow reduction and reperfusion flow, preload, afterload, and inotropic stimulation were the same during agranulocytic and granulocytopenic perfusion. During agranulocytic perfusion, stunning did not occur (greater than 100% of preischemic function during reperfusion), but when the filters were removed and about 10% of the normal granulocyte count was present, stunning occurred with only 76% return of function at 60 minutes of reperfusion (p less than 0.01). A second series of studied animals with extracorporeal perfusion and granulocyte replete perfusion all had less than 75% return of regional function, indicating that the agranulocytic perfusion and not the extracorporeal aspects of the experiment prevented stunning. We conclude that granulocytes are the direct source of the injury in stunned myocardium and apparently the main source of superoxide in the 15-minute ischemia dog model. Other possible granulocyte-related mechanisms of reperfusion injury include capillary no-reflow, causing microvascular ischemia and degranulation leading to enzyme-induced damage. PMID- 3038367 TI - Proarrhythmic effects of an oxygen-derived free radical generating system on action potentials recorded from guinea pig ventricular myocardium: a possible cause of reperfusion-induced arrhythmias. AB - Standard microelectrode techniques were used to study the effects of a free radical generating system on action potentials recorded from guinea pig ventricular myocardium. Free radicals were generated by mixing xanthine oxidase (0.02-0.04 mu/ml) with the superfusate-modified Locke's solution containing purine 2.3 mM. The system was validated by demonstrating that it could reduce cytochrome C at a rate of 15.9 +/- 1.5 mol/l/min. This rate was decreased to 3.0 +/- 0.3 (p less than 0.001) in the presence of superoxide dismutase (12 mg/100 ml), and the reaction was absent if xanthine oxidase and purine were premixed for 60 minutes prior to adding cytochrome C. Superfusion of guinea pig ventricular strips with the free radical generating system (20-30 minutes) resulted in a highly significant reduction in resting potential from -79.3 +/- 1.8 mV to -70.9 +/- 1.4 mV (p less than 0.0001, n = 6) and in action potential amplitude from 110.9 +/- 2.2 mV to 101.7 +/- 4.0 mV (p less than 0.0001). There was an accompanying fall in maximum rate of depolarization (Vmax) from 254.1 +/- 17.7 V/sec to 207.1 +/- 18.6 V/sec (p less than 0.01) and no significant change in action potential duration. These changes were accompanied by spontaneous activity in 3 of 6 preparations and reversed after 20-30 minutes washing in Locke's solution. They were largely abolished by adding superoxide dismutase (12 mg/100 ml) to the superfusate and completely absent if the xanthine oxidase and purine were premixed for 60 minutes before superfusing the myocardium. We conclude that the phenomena observed may contribute to the genesis of reperfusion arrhythmias. PMID- 3038368 TI - Desensitization of postjunctional alpha 1- and alpha 2-adrenergic receptor mediated vasopressor responses in rat harboring pheochromocytoma. AB - Prolonged stimulation of tissues by adrenergic agonists may lead to diminished responsiveness of the tissues to subsequent activation by catecholamines; this phenomenon has been termed desensitization or tachyphylaxis. We have examined the in vivo consequences of prolonged stimulation of vascular alpha-adrenergic receptors in rats harboring pheochromocytoma, a tumor that secretes catecholamines. In both early (3-4 weeks after implantation) and late (6-7 weeks after implantation) stages of tumor development, New England Deaconess Hospital rats with transplanted pheochromocytomas developed hypertension and tachycardia and had plasma dopamine and norepinephrine concentrations markedly greater than controls. In both these stages of pheochromocytoma, pressor responses to several vasoconstrictors were examined after pithing. Rats with the tumor were found to become progressively subsensitive to alpha-adrenergic agonists. In the early phase of pheochromocytoma, loss in sensitivity was found for both alpha 1- and alpha 2-adrenergic agonists, whereas responsiveness to the nonadrenergic vasoconstrictors Arg-vasopressin and angiotensin-II was intact (homologous desensitization). However, in the later stage of pheochromocytoma, pressor responses to all these vasoconstrictive agents and also to stimulation of the complex sympathetic outflow were found to be subsensitive (heterologous desensitization). In plasma membranes prepared from mesenteric arteries of early stage tumor-bearing rats, [3H]prazosin binding sites were significantly decreased to 150 +/- 12 fmol/mg vs. 234 +/- 19 fmol/mg in controls. [3H]Yohimbine binding sites were not significantly altered. Our results show that both postjunctional alpha 1- and alpha 2-adrenergic receptor-mediated vasopressor responses can be specifically attenuated in the presence of chronically elevated endogenous catecholamine levels produced by pheochromocytoma and that each alpha-receptor subtype may be differently regulated in the development of desensitization. PMID- 3038369 TI - Denervation supersensitivity of refractoriness in noninfarcted areas apical to transmural myocardial infarction. AB - Denervation supersensitivity was demonstrated in anesthetized dogs 5 to 10 days after transmural myocardial infarction produced by latex embolization of a diagonal branch of the left anterior descending coronary artery. Sympathetic efferent denervation in noninfarcted myocardium apical to the infarction was demonstrated by a 90% depletion of myocardial norepinephrine content in the apical (45 +/- 15 pg norepinephrine/g tissue) vs basal (437 +/- 76 pg/g tissue) regions and by the lack of effective refractory period (ERP) shortening during bilateral ansae subclaviae stimulation in 34% of sites apical to the infarction. Supersensitivity in the area apical to the infarction was manifested by an exaggerated shortening of the ERP during both norepinephrine and isoproterenol infusions, with an upward and leftward shift in the dose-response curves in the apical vs basal regions (p less than .001). The cellular mechanism for denervation supersensitivity did not involve detectable changes in the beta adrenergic receptor adenylate cyclase system. There was no difference in the density of beta-adrenergic receptors ([125I]-cyanopindolol) in the apical (268.6 +/- 22.7 fmol/mg protein) vs the basal (253.5 +/- 24.8 fmol/mg protein) regions. Adenylate cyclase activity stimulated by guanosine triphosphate plus isoproterenol was slightly greater in the apical (58.7 +/- 17.4%) than in the basal (49.6 +/- 10.9%) region, but this difference did not reach statistical significance (p = .068). Muscarinic modulation of beta-receptor coupling (oxotremorine attenuation of guanosine triphosphate plus isoproterenol-stimulated adenylate cyclase activity) also was not significantly different at the apical (31.6 +/- 17.5% inhibition) and basal (21.4 +/- 20.9% inhibition) sites. These data show that a transmural myocardial infarction produces denervation supersensitivity in areas apical to the infarction, but in this preparation no differences in the total number or a redistribution of beta-adrenergic receptors or adenylate cyclase activity were detected. PMID- 3038370 TI - The path to myocardial salvage by thrombolytic therapy. PMID- 3038372 TI - Use of thallium-technetium parathyroid scans. PMID- 3038371 TI - Radionuclide methods for evaluating the results of thrombolytic therapy. AB - In summary, a variety of nuclear techniques may be used to investigate the effects of thrombolytic therapy and myocardial reperfusion. Assessments of global and regional ventricular function, myocardial perfusion, and metabolic integrity are available and appear to add substantially to conventional assessment. Timing of studies appears to be critical. Complementary data can be obtained in both the acute and convalescent phase of myocardial infarction. PMID- 3038373 TI - Penicillin in milk--its importance in urticaria. AB - Fifty patients with recurrent urticaria were tested by means of the RAST test for penicillin allergy. Fifteen patients had positive reactions and of these, thirteen received provocation tests with 0.1 U/ml of penicillin in milk. Four had definite positive reactions, three doubtful reactions and six had no reaction. Although there has been improvement in the purity of milk, penicillin residues remain a potential cause of urticaria even in very low amounts and could have contributed to the urticaria in at least 8% of our patients. Veterinary use of antibiotics and food quality should be strictly regulated to prevent contamination of our diet. PMID- 3038374 TI - The role of arterial mineralocorticoid receptors in the mechanism of hypertension: findings and hypothesis. AB - Alterations in electrolyte transport across cell membrane of vascular smooth muscle (VSM) and changes in hemodynamics [increased extracellular fluid volume (ECFV) and cardiac output (C.O.)] have been implicated in the pathogenetic mechanisms of both mineralocorticoid-induced hypertension (MH) and essential hypertension (EH). We have previously found that mineralocorticoids (MC) can act directly on arterial wall by means of a receptor-mediated mechanism, and have postulated that this mechanism is of critical importance in the increased reactivity of VSM to vasoconstrictive stimuli in MH. We now present evidence that a MC-antagonist at the MC-receptor level, progesterone, prevents induction of changes in VSM cell-membrane permeability to Na+ by MC, and development of hypertension. This study has been carried out on rabbits made hypertensive by s.c. implantation of silastic rubber strips impregnated with 11 desoxycorticosterone (the inducer) and/or 50 times that amount of progesterone (the anti-inducer). We hypothesize that the VSM cell-membrane defect (MC-induced in MH and congenital in EH) initiates two separate sequences of biochemical events. One leads, in early stages of hypertension, to expansion of ECFV, increase in C.O., myogenic vasoconstriction and hypertension. When kidney function matures, hypertension recedes. The second sequence of events leads to hypertension via an increase in [Na]i of VSM, leading to an increase in [Ca]i, and an increased contractility of VSM. This hypertension persists. The two sequences are concomitant but independent of each other. PMID- 3038375 TI - Collagenase activity in colonic mucosa during inflammatory bowel disease. AB - A simple test for collagenase activity was performed on colonic mucosa specimens of 35 patients with inflammatory bowel disease, 7 patients with carcinoma of the colon, 3 with benign polyps, and 34 normal subjects. Increased collagenase activity was present in 94.2% of the specimens taken from the inflamed mucosa and 71.4% of those obtained from colonic carcinoma, as compared to 8.8% of the control group. PMID- 3038376 TI - Immobilized purified folate-binding protein: binding characteristics and use for quantifying folate in erythrocytes. AB - Purified folate-binding protein from cow's milk was immobilized on monodisperse polymer particles (Dynospheres) activated by rho-toluenesulfonyl chloride. Leakage from the spheres was less than 0.1%, and the binding properties were similar to those of the soluble protein with regard to dissociation, pH optimum for binding pteroylglutamic acid, and specificity for binding various folate derivatives. We used the immobilized folate-binding protein as binding protein in an isotope-dilution assay for quantifying folate in erythrocytes. The detection limit was 50 nmol/L and the CV over a six-month period was 2.3% (means = 1.25 mumol/L, n = 15). The reference interval, for folate measured in erythrocytes of 43 blood donors, was 0.4-1.5 mumol/L. PMID- 3038378 TI - Reagent that restores galactose-1-phosphate uridylyltransferase activity in dry blood spots. PMID- 3038377 TI - Automated quantification of choriogonadotropin: analytical correlation between serum and urine with creatinine correction. AB - We evaluated the Boehringer Mannheim Diagnostics ES 600 automated immunoassay system for measuring choriogonadotropin (hCG) in human serum, plasma, and urine. The ES 600, an automated sample-selective, multibatch analyzer, is based on immunoenzymometric coated-tube technology. Interassay CVs for assay of both serum and urine ranged from 2% to 16%. The analytical sensitivity was 0.5 int. unit/L. Results for serum correlated well with qualitative "Icon" and quantitative "Tandem" assays (Hybritech). In addition, results for serum and urinary hCG correlated well when the latter was expressed in terms of creatinine excretion. These results suggest that determination of hCG in urine can be correlated with those in serum for use as a marker for chorionic tumors or for screening abnormal pregnancies, where the rate of hCG production may be important. PMID- 3038379 TI - Why are there different reference intervals for the kinetic angiotensin converting enzyme assay? PMID- 3038380 TI - Statistical significance of paraoxonase activity in a control group of children and a group of children of parents who had had early myocardial. PMID- 3038381 TI - Agarose gel isoelectrofocusing of UDP-galactose pyrophosphorylase and galactose-1 phosphate uridyltransferase. Developmental aspect of UDP-galactose pyrophosphorylase. AB - The uridine diphosphogalactose pyrophosphorylase activity has been determined in human adult and fetal tissues as well as blood of various ages by measurement of UDP-galactose production from gal-1-p and UTP. The highest activity was found from adult liver in which the specific activity was about 5% of the gal-1-p uridyltransferase activity. In general adult tissues had a somewhat higher activity than the corresponding fetal tissues except erythrocytes in which fetuses had a 5-10 times higher activity than adults. From normal blood the pyrophosphorylase activity in erythrocytes decreased with age, but in the case of galactosemia the decrease with age was not distinct. According to agarose gel isoelectrofocusing studies, at least two isozyme forms for UDP-galactose pyrophosphorylase exist with the activity bands between pH 6.0-6.15. The patterns of AGIF bands of pyrophosphorylase varied according to the age of the samples, suggesting the development of the isozyme forms of pyrophosphorylase to be age dependent. Uridyltransferase, on the other hand, resolved into multiple bands between pH 5.1-5.6 on agarose gels and the patterns varied according to the variants but not to the age. Significance of the decrease in the pyrophosphorylase activity in erythrocytes with age as well as of the difference in AGIF bands between normal and the galactosemic were discussed with regard to the pathology of classical galactosemia. PMID- 3038382 TI - Biochemistry of human converting enzyme. AB - Angiotensin converting enzyme (ACE, EC 3.4.15.1) was purified to homogeneity from human kidney and its specific antibody was raised in the rabbit. The antibody cross-reacted equally with human enzymes from kidney, lung, intestine, plasma and urine. Immunofluorescent and immunoelectron microscopic observation indicated that the enzyme was located on the plasma membrane and micropinocytic vesicles at the luminal site of vascular endothelium in the lung. It was also present on the brush border, intercellular and basal infolding membranes of proximal tubular epithelium, but was not detected on the distal tubular epithelium or vascular endothelium in the kidney. ACE was demonstrated immunocytologically in human alveolar macrophages and renal carcinoma tissues. The carcinoma tissue contained a possible isoenzyme of ACE differing in part immunologically from the enzyme of normal kidney. PMID- 3038383 TI - Proceedings of the International Symposium on Pharmacological and Clinical Evaluation of Converting Enzyme Inhibitors. Kyoto, Japan, July 27-29, 1986. PMID- 3038384 TI - Pulmonary and testicular angiotensin-converting isoenzymes. AB - A variant of angiotensin-converting enzyme occurs in (male) germinal cells. This testicular isozyme is catalytically similar to the widespread pulmonary-type isozyme, but contains a shorter polypeptide chain and does not appear until puberty. The two proteins differ at their NH2- and COOH-termini, but share many tryptic peptides. All antigenic determinants of the testicular form are represented in the pulmonary molecule whereas the latter contains determinants unrelated to catalysis which are lacking in the testicular species. The data indicate that the testicular isozyme corresponds closely to an internal part of the pulmonary polypeptide which includes its active site. The structural and developmental differences between the two polypeptides are pretranslationally determined since they are demonstrable in a cell-free system programmed by the appropriate mRNAs. Characterization of the molecular mechanisms responsible for the relationship of these isozymes may yield useful information regarding cell specific protein expression. PMID- 3038385 TI - Angiotensin-converting enzyme inhibitors: accomplishments and challenges. PMID- 3038386 TI - No substrate specificity of converting enzyme for N-terminal substituted angiotensin I in man. AB - In 5 normal men sarcosine1-angiotensin II (Sar1-ANG II) (Exp. 1) and sarcosine1 angiotensin I (Sar1-ANG I) (Exp. 2) infused iv at a rate of 5 pmol/kg X min from 0900 h to 0930 h caused the same degree of rise in blood pressure (BP). But 100 mg of captopril given orally at 0800 h (Exp. 3) completely abolished the BP rise by Sar1-ANG I. In Exps. 1 and 2 plasma renin activity (PRA) decreased and plasma aldosterone (PA) increased after the infusions. In Exp. 3 PRA increased markedly and PA decreased 60 min after captopril, and at 30 min of Sar1-ANG I infusion PRA decreased to the pre-captopril level despite no BP change but PA was kept at the pre-infusion level. Hence, substrate specificity of converting enzyme previously demonstrated for N-terminal deleted ANG I was not shown for N-terminal substituted ANG I in man because the conversion of Sar1-ANG I to Sar1-ANG II was 100%. Sar1-ANG I may possibly inhibit renin release in normal men. PMID- 3038387 TI - Biochemical characterization of angiotensin-converting enzyme in human neuroblastoma tissue. AB - High activity of angiotensin-converting enzyme was demonstrated in human neuroblastoma tissue. This activity required the presence of chloride ion and was almost completely inhibited by a specific converting enzyme inhibitor captopril (10 nM), indicating that the activity measured is indeed angiotensin-converting enzyme. Furthermore, the biochemical features of the enzyme were closely similar to the well-known properties of human lung converting enzyme, such as molecular weight (290,000), optimum pH (8.0-8.5), the presence of glycoprotein residues, and dependence on chloride ion concentration. These results provide definitive evidence for the presence of true angiotensin-converting enzyme in human neuroblastoma tissue. PMID- 3038388 TI - Pharmacology of converting enzyme inhibitors. AB - Angiotensin converting enzyme inhibitors were developed to prevent the in vivo generation of angiotensin II and thereby to reduce peripheral vasoconstriction. However, these compounds exert some additional effects that may or may not be angiotensin dependent. These include potential sodium diuresis, bradykinin accumulation, prostaglandin release, blunting of sympathetic activity, parasympathomimetic actions, central effects, redistribution of blood flow toward some particularly important organs. Only the comprehensive assessment of the many complex interactions that exist between the renin-angiotensin and several other regulatory systems reveals the complete therapeutic profile of this class of pharmacologic agents. PMID- 3038389 TI - Changes in arterial distensibility produced by converting enzyme inhibitors in hypertensive humans. AB - Converting enzyme inhibitors enhance arterial compliance in hypertensive humans. The enhancement is due not only to an increase in arterial diameter and volume but also to an increase in arterial distensibility. The latter effect reflects the drug action on arterial smooth muscle tone rather than the lower stretch due to the blood pressure reduction. The improvement in the buffering function of large arteries in hypertensives may contribute to produce a more important decrease in systolic pressure and a better reversion of cardiac hypertrophy. PMID- 3038390 TI - Acute and subacute hemodynamic effects of enalaprilat, milrinone and combination therapy in rats with chronic left ventricular dysfunction. AB - A model of congestive heart failure (CHF) was produced in rats approximately 76 days following surgical occlusion of the left coronary artery. In rats with healed myocardial infarction (MI size = 45% LV), hemodynamic variables were predictably elevated (LVEDP greater than 20 mm Hg) or depressed LV dP/dtmax (5200 mm Hg/sec). The hemodynamic response (MAP, HR, LVEDP, and dP/dtmax) to intravenous infusion of Mil (54 to 347 micrograms/kg) was measured on two occasions, separated by a 7-12 day period of oral treatment (2 mg/kg/day). Enalaprilat was tested in a similar design with the infusion phase (70 micrograms/kg, total dose) separated by oral enalapril (Enal), 1 mg/kg/day. The hemodynamic response to Mil was also examined in rats treated with the ACE inhibitor. At low doses, Mil modestly elevated HR (+17 beats/min) and dose dependently increased (P less than 0.05) LV contractility by approximately 25%. Higher doses of Mil reduced preload (LVEDP) and afterload (MAP). Oral Mil produced little hemodynamic improvement except modest elevation (+9%) in LV contractility. There was no evidence of tachyphylaxis to i.v. Mil. In contrast, enalaprilat reduced MAP and preload without altering HR or contractility, effects observed after oral treatment. In the presence of the ACE inhibitor, Mil's hemodynamic actions were not enhanced. These experiments demonstrate that ACE inhibition improves ventricular performance by reducing preload and afterload. In this model, Mil improves performance by a direct inotropic mechanism as well as a reduction in afterload. PMID- 3038391 TI - Angiotensin converting enzyme inhibition in plasma and tissues. AB - Angiotensin converting enzyme (ACE) and the ACE inhibitor lisinopril were measured in patients with renal impairment, by both radioinhibitor 125I MK351A binding studies, and by radioimmunoassay. Plasma concentration of lisinopril estimated by radioinhibitor binding displacement correlated closely with that measured by radioimmunoassay. Plateau lisinopril concentration in 8 patients with varying degrees of renal failure treated with 5 mg lisinopril per day for 1 week, was inversely related to renal function. Plasma lisinopril concentrations of 30 70 ng/ml were required for 50% inhibition of plasma ACE activity in vivo. Acute studies in the rat showed inhibition of ACE in different tissues had different time courses. These observations suggest that 125I MK351A binding studies in tissues will be useful in establishing the pharmacokinetic and pharmacodynamic profiles of newer ACE inhibitors, and may help delineate the contribution of ACE in different tissues to cardiovascular homeostasis. PMID- 3038392 TI - Comparison of chronic inhibition of renin and converting enzyme in the marmoset. AB - The primate-specific renin inhibitor CGP 29287 (30 mg/kg/d, n = 5) or the converting-enzyme inhibitor CGS 14831 (30 mg/kg/d, n = 8) were administered by continuous intraperitoneal infusion via osmotic minipumps to normotensive marmosets fed a low salt diet. CGP 29287 and CGS 14831 induced a similar reduction in blood pressure after 2 days of administration (-22 +/- 6, SEM and 24 +/- 6 mmHg respectively). The hypotensive response persisted at 7 days (-20 +/ 3 and -22 +/- 8 mmHg respectively). Despite the fall in blood pressure, heart rate was not changed after either inhibitor. Blood pressure and heart rate remained stable in control marmosets that received vehicle only (0.9% saline). Plasma renin activity was inhibited after CGP 29287 (100% at day 2 and 75% at day 7) and increased after CGS 14831 (4 and 3 fold on days 2 and 7 respectively). Total plasma immunoreactive renin was increased to a similar extent after both inhibitors (approximately 5 fold on days 2 and 7). These findings show that a similar hypotensive response is induced in sodium-depleted primates after chronic inhibition of renin or converting enzyme. Thus the fall in blood pressure after chronic treatment with either inhibitor appears to be mainly due to the blockade of the renin-angiotensin system and the consequent reduction in endogenous angiotensin II formation. PMID- 3038393 TI - Difference in response of vascular angiotensin converting enzyme activity to cilazapril in SHR. AB - Angiotensin I converting enzyme (ACE) activity was found in all rat arteries and veins examined, considerably varied among these vessels, and usually higher in arteries than in corresponding veins except for femoral and subclavian pairs. Decrease of vascular ACE activity in response to cilazapril at hypotensive doses varied in vessels and was not dependent on their ACE activity levels. PMID- 3038394 TI - Synergism of CE-inhibition and diuresis. A study with ramipril, piretanide and HCT in SHR. PMID- 3038395 TI - Perindopril, a new angiotensin converting enzyme inhibitor--clinical pharmacological studies in healthy subjects. AB - Perindopril is a new angiotensin converting enzyme (ACE) inhibitor which is activated after hydrolysis in vivo to a diacid (S9780). Oral administration of perindopril (1-16 mg) to groups of 6 healthy males led to a long lasting and dose elated inhibition of plasma ACE and rises in plasma renin activity with no evidence of accumulation of drug or effect. At higher doses there was a modest fall in blood pressure. S9780 given intravenously in doses of 1,2 or 4 mg caused an immediate inhibition of plasma ACE. The concentration of S9780 in plasma which led to 50% inhibition of plasma ACE was 1.55 +/- 1.14 ng/ml (mean +/- S.D.). Clinical trials with 2-8 mg once daily in essential hypertension are currently in progress. PMID- 3038396 TI - Protective effect of angiotensin converting enzyme inhibitors (CEI): captopril and perindopril on vulnerability to ventricular fibrillation during myocardial ischemia and reperfusion in rat. AB - The purpose of the present study was to evaluate the effects of two CEI: captopril and perindopril on reperfusion arrhythmias and noradrenaline 3H (NA3H) release in the isolated rat heart on arrhythmias in pentobarbitone anaesthetized rats subject to left coronary artery ligation. In vitro, in control preparations, reperfusion after 10 min of local ischemia produced by coronary ligation was accompanied by a sudden release of NA3H in coronary flow and long lasting ventricular arrhythmias. Reperfusion arrhythmias were prevented by perfusion medium containing a high dose of CEI and CEI did not change the patterns of NA3H release. In vivo, mortality due to ventricular fibrillation was significantly reduced in rats pretreated with captopril or perindopril 15 min prior to coronary ligation. CEI reduced the severity of ventricular tachycardia (VT) and fibrillation (VF). PMID- 3038397 TI - Effects of chronic treatment with ramipril, a new ACE blocking agent, on presynaptic sympathetic nervous system of SHR. AB - Acute administration of captopril has been reported to decrease noradrenaline (NA) release from the sympathetic nerves. In this study the chronic effects of ramipril on the sympathetic nervous system of male spontaneously hypertensive rats (SHR) have been investigated and compared to those of captopril and enalapril. As parameters for catecholamine biosynthesis and storage, the activity of tyrosine hydroxylase and the catecholamine content of the hearts and the adrenal medulla were measured by HPLC in treated and control SHR. To assess sympathetic outflow plasma NA and adrenaline (A) levels were determined during preganglionic stimulation (PS) of the spinal cord. Under none of these drugs could differences be observed between the treated and control animals, neither in the biosynthesis and storage of catecholamines in the heart and adrenal medulla nor in the sympathetic outflow. However, the dose response curves of blood pressure vs PS were significantly shifted to the right when ACE-inhibitors were administered, most strongly by ramipril. In view of the unaltered presynaptic sympathetic function long term treatment with ACE-inhibitors is suggested to increase bradykinin and angiotensin I. Bradykinin and angiotensin I are capable of releasing NA and A from the adrenal medulla, like angiotensin II. PMID- 3038398 TI - Effects of captopril on neurosecretion and vascular responsiveness in hypertension. AB - The purpose of the present study was to investigate the mechanisms of hypotensive action of an angiotensin converting enzyme inhibitor (captopril) in hypertension. In perfused mesenteric vasculatures from spontaneously hypertensive rats (SHR) and normotensive Wistar kyoto rats (WKY), the effects of captopril on the vascular responsiveness and norepinephrine overflow from the adrenergic nerve endings were examined. The vasoconstrictor responses and norepinephrine overflow during the electrical nerve stimulation was significantly enhanced in SHR compared with WKY. Captopril reduced not only vasoconstrictor responses but also norepinephrine overflow during the nerve stimulation in a dose-dependent fashion. The suppressions of these responses by captopril were significantly greater in SHR than in WKY. These results demonstrate that captopril could affect the presynaptic site of the resistance vessels and cause a decrease in electrically stimulated norepinephrine overflow from the adrenergic nerve endings. The marked reduction of the pressor responses and norepinephrine overflow to nerve stimulation by captopril in the SHR suggests that the renin-angiotensin system in the vascular beds in enhanced in this model of hypertension. PMID- 3038399 TI - Monocrotaline-induced cardiopulmonary injury in rats: modification by thiol and nonthiol ACE inhibitors. PMID- 3038400 TI - Hypotensive effects of the renin inhibitor (RI-78) and the converting enzyme inhibitor (teprotide) in conscious monkeys. AB - Effects of the pentapeptide renin inhibitor (RI-78; Phe(4Cl)-Phe-Val-Tyr-Lys-NH2) and the angiotensin converting enzyme (ACE) inhibitor (teprotide) on mean arterial pressure (MAP) were examined in conscious monkeys (M. fascicularis). In salt depleted normotensive monkeys with a MAP of 95 +/- 4 mmHg and plasma renin activity (PRA) of 15.9 +/- 2.7 ngAI/ml/h, a bolus injection of a dose of 375 micrograms/kg of RI-78 caused a prompt hypotensive effect. Maximal hypotensive action was seen within 1 min, and MAP returned to the basal level within 15 min. With this dose, MAP was reduced by 20 +/- 6 mmHg. Teprotide (1 mg/kg) decreased MAP and reached a nadir after 13 min. There was no significant difference between maximal hypotensive responses seen with RI-78 (375 micrograms/kg) and with teprotide (1 mg/kg). Hypotensive effects of RI-78 and teprotide were also examined in acute renal hypertensive monkeys with a MAP of 125 +/- 5 mmHg and a PRA of 27.1 +/- 5.7 ngAI/ml/h. Again, similar hypotensive effects were observed. We conclude that antihypertensive effect of RI-78 is comparable to that seen with teprotide. PMID- 3038401 TI - Antihypertensive action of angiotensin-I converting enzyme inhibitors in the kidney. AB - In order to elucidate the antihypertensive action of angiotensin-I converting enzyme inhibitors (CEIs) in the kidney, intrarenal (IR) administration of CEIs was performed in norepinephrine (NE) or angiotensin-II (AII) induced acute hypertension in conscious unrestrained rabbits. IR administration of the two kinds of CEIs, captopril (5 mg/kg) and MK-422 (1 mg/kg), dose which caused no significant effect when injected intravenously, elicited a prompt and marked depressor action in NE but not in AII induced hypertension. In addition, IR infusion of the angiotensin-II analogue, saralasin (2 and 10 micrograms/kg/min) showed no depressor effect in NE induced acute hypertension. These results indicate that the kidney plays an important role in the depressor action of CEIs in NE but not in AII induced hypertension and this effect may not be related to the IR renin-angiotensin system. PMID- 3038403 TI - Regulation of prostacyclin generation by angiotensin converting enzyme related substances in cultured human vascular endothelial cells. AB - The regulation of prostacyclin (PGI2) generation by angiotensin I-converting enzyme (ACE) related substances was investigated using cultured human vascular endothelial cells. Angiotensin I (AI) or bradykinin (BK) increased PGI2 generation and ACE activity, while the ACE inhibitor, captopril decreased both of them, and angiotensin II (AII) did not show any effect. The increasing rate of PGI2 generation induced by AI or BK was not affected by the pretreatment with captopril. These results suggest that the accumulation of AI or BK via the inhibition of ACE by captopril did not cause the enhancement of PGI2 generation. Rather, it was proposed that the enhanced PGI2 generation by AI or BK might be regulated by ACE activation derived from these substances, as an autoregulation mechanism. PMID- 3038402 TI - Effects of angiotensin converting enzyme inhibitors MK 421, SA 446 and captopril on renal prostaglandin E and kinins in conscious normotensive rats. AB - Angiotensin converting enzyme inhibition by MK 421, SA 446 or captopril (6 mg/kg/day ip) for up to 6 days induced significant fall in systolic blood pressure and plasma angiotensin II concentration. The hypotensive effect was greater in sodium depleted rats than in sodium repleted rats. The hypotensive effect was also accompanied by increased excretion of urinary prostaglandin E2, however the levels of urinary prostaglandin E2 in sodium repleted rats were not different from those in sodium depleted rats. Urinary kinin excretion was increased during infusion of MK 421, SA 446 or captopril in sodium depleted rats, whereas no significant change was found in sodium repleted rats. Thus the present results suggest that renal prostaglandin E system may not be essential for the hypotensive effect of these inhibitors. In addition, the greater hypotensive effect of these inhibitors in sodium depleted rats may be in part due to stimulated renal kinin system. PMID- 3038404 TI - Prevention of renal damage and decrease of urinary kinins excretion by chronic treatments with enalapril and captopril in stroke-prone spontaneously hypertensive rats. PMID- 3038405 TI - Tissue converting enzyme inhibition and cardiovascular effects of converting enzyme inhibitors. AB - Local renin-angiotensin systems have been described in organ controlling cardiovascular function. Following oral administration of converting enzyme (CE) inhibitors the enzyme was inhibited not only in lung vascular endothelium and blood, but also in the heart, kidney, vascular wall, brain and other organs. Evidence for a contribution of tissue CE inhibition to the cardiovascular actions of CE inhibitors is provided by the antihypertensive effect of brain CE inhibition in spontaneously hypertensive rats, by the concomitant persistence of blood pressure decrease and CE inhibition in vascular wall and kidney after chronic oral CE inhibitor treatment, and by the marked effects of oral CE inhibitor pretreatment on cardiac function in isolated rat hearts. Local inhibition of tissue renin-angiotensin systems may therefore be an important factor involved in the beneficial effects of CE inhibitors in cardiovascular diseases such as arterial hypertension, congestive heart failure, and cardiac arrhythmias. PMID- 3038406 TI - Heart and vascular wall as targets for tissue converting enzyme inhibition. PMID- 3038407 TI - Possible role of vascular angiotensin converting enzyme in the genesis of hypertension. AB - This study was designed to evaluate changes in angiotensin converting enzyme (ACE) activity in 2-kidney, 1-clip renal hypertensive dogs. Blood pressure increased and remained elevated for eight months after partial clamping of the left renal artery. Plasma renin activity was increased for one month after surgery, but then returned to the pre-clamping level. SA-446, a potent ACE inhibitor, reduced blood pressure in 8 M-hypertensive dogs. Plasma ACE activities was not altered during the course of hypertension, but vascular ACE activities in the pulmonary and mesenteric arteries and the aorta were significantly increased in 8 M-hypertensive dogs. These results may indicate that ACE plays an important role in the maintenance of chronic phase of 2-kidney, 1-clip renal hypertension and that suppression of vascular ACE activities might be a mechanism underlying the hypotensive effect of ACE inhibitors. PMID- 3038408 TI - Contribution of renal angiotensin converting enzyme (ACE) to blood pressure regulation: possible role of brush border ACE. AB - The role of renal angiotensin converting enzyme (ACE) in blood pressure regulation is not well understood. In our studies, both acute and chronic treatment of hypertensive rats SHR and SHRSP with ACE inhibitors Enalapril and SA446 had a blood pressure lowering effect that coincided with an inhibition of renal cortical and aortic ACE, but not plasma ACE. Further, ACE activities in the renal cortex and aorta were found to increase with aging of the SHRSP, therefore concomitantly with hypertension development. In the kidney, brush border membranes (BBM) contained abundant ACE. We found that the activities of ACE in the renal cortex closely correlated to the activities in isolated BBM, in Wistar Kyoto rats and in the SHRSP. Thus, renal cortical ACE activity and blood pressure correlated in cases of ACE inhibition and hypertension development. Since the ACE activity in the renal cortex appeared to reflect the enzyme activity in BBM, the brush border ACE may have to be taken into account, in view of the relationship between renal ACE and blood pressure. PMID- 3038409 TI - Distribution of angiotensin converting enzyme in sheep hypothalamus and medulla oblongata visualized by in vitro autoradiography. AB - In vitro autoradiographic mapping of angiotensin converting enzyme (ACE) in sheep brain using the specific ACE inhibitor, 125I-351A, revealed very high densities of binding in large blood vessels and choroid plexus. In the a very high density of labelling occurred in the organum vasculosum of the lamina terminalis and median eminence and a high density in the subfornical organ and moderate density in supraoptic, suprachiasmatic, arcuate and paraventricular nuclei. All fiber tracts were unlabelled. In the medulla oblongata, a very high density of binding was detected in the area postrema and a high density in the nucleus of the solitary tract and dorsal motor nucleus of the vagus; a moderate density was found in the substantia gelatinosa of the spinal tract and the inferior olivary nucleus. PMID- 3038410 TI - Central and peripheral inhibition of angiotensin converting enzyme (ACE) in the SHR: correlation with the antihypertensive activity of ACE inhibitors. AB - A series of five structurally distinct ACE inhibitors were evaluated for their ability to inhibit tissue ACE activity in the SHR after oral administration. In the first series of experiments, the ACE inhibitors captopril, enalapril, pentopril, CGS 14824A and CGS 16617 were given to groups of SHR at doses that produced a 15 to 20 mm Hg reduction in blood pressure within 1 hour. Under these conditions of dose and time, only captopril significantly inhibited brain ACE activity (43%), whereas inhibition of serum ACE activity ranged from 72% to 99% with these agents. Inhibition of aortic ACE activity ranged from 54% to 87%, and lung ACE inhibition varied from 50% to 83%. In the second series of experiments, SHR were administered higher doses of each ACE inhibitor such that these compounds produced peak reductions in blood pressure (-25 mm Hg to -33 mm Hg) within a range of 2 to 6 hours. When tissue ACE activity was measured at the time corresponding to peak reduction in blood pressure, all five ACE inhibitors produced a significant inhibition of brain ACE activity ranging from 21% to 76%. Serum ACE activity was almost completely inhibited by these agents, with the exception of captopril (62% inhibition). The inhibition of aortic ACE activity ranged from 79% to 99%, while the inhibition of lung ACE activity did not increase under these conditions. These data suggest that ACE inhibitors may exert their maximal antihypertensive effects by inhibiting ACE in vascular tissues and brain. PMID- 3038411 TI - Distribution of kininase activity along the rabbit nephron. AB - The distribution of kininases along microdissected nephron segments was studied. Single nephrons from collagenase treated rabbit kidney were microdissected and divided into following 9 segments: glomerulus; early proximal tubule; middle proximal tubule; late proximal tubule; cortical thick ascending limb; distal convoluted tubule; connecting tubule; cortical collecting tubule; medullary collecting tubule. Kininase activities in these nephron segments were measured with or without kininase II inhibitor. Kininase II and other peptidases were mainly localized in glomeruli and whole part of the proximal tubules. PMID- 3038412 TI - Effects of chronic administration of angiotensin converting enzyme (ACE) inhibitors on blood pressure and tissue ACE activity in the SHR. PMID- 3038413 TI - Angiotensin converting enzyme in sarcoidosis and in silicosis. AB - Angiotensin converting enzyme (ACE) activities of bronchoalveolar lavage fluid (BALF) and serum in patients with sarcoidosis and with silicosis were measured. Serum ACE was measured by enzymic assay and radioimmunoassay. There was a close relationship between ACE activity and content (r = 0.78). Serum ACE activities in patients with active sarcoidosis (37.5 +/- 11.1 nmol/min/ml, mean +/- SD) and with silicosis (25.5 +/- 9.3) were significantly elevated from the control (18.6 +/- 6.0). BALF ACE activities in the control, patients with active sarcoidosis and with silicosis were 0.23 +/- 0.19 nmol/min/ml, 0.94 +/- 0.97 and 0.38 +/- 0.05, respectively. BALF ACE in patients with active sarcoidosis and with silicosis were significantly different from the control. When BALF ACE was corrected by the cell count of alveolar macrophage (per 10(6) cells), activity was significantly different from control only in the patients with sarcoidosis. Moreover, only the alveolar macrophages in sarcoidosis were stained by immunofluorescence and immunocytochemistry using rabbit antihuman ACE antibody. Induction of ACE in alveolar macrophage might have an important role for the activity or progression of sarcoidosis. PMID- 3038414 TI - The clinical use of angiotensin converting enzyme inhibitors in hypertension and cardiac failure. AB - The renin-angiotensin system has a range of physiological actions concerned with the control of the circulation. Angiotensin II has both an immediate and a delayed pressor effect, it stimulates the secretion of aldosterone and antidiuretic hormone, promotes thirst, stimulates the sympathetic nervous system at various sites while inhibiting vagal tone, and has a range of direct effects on the kidney. Several aspects of this range of actions can become deranged in a number of forms of hypertension as well as in congestive cardiac failure. Hence much effort has been directed in recent years to the development of agents designed to interfere with the renin-angiotensin system and to apply these clinically in the treatment of hypertension and congestive cardiac failure. Orally active converting enzyme inhibitors are of proven benefit not only in renovascular hypertension, but also, when combined with loop diuretics, in the treatment of intractable hypertension as well as, both alone and in combination with thiazide diuretics, in the treatment of essential hypertension. In congestive cardiac failure controlled trials have shown that converting enzyme inhibitors can improve exercise tolerance while diminishing lassitude, correct potassium deficiency and limit ventricular arrhythmias. Energetic efforts are being made to develop orally active inhibitors of the enzyme renin itself, since these would be more specific in action than the presently available and very successful converting enzyme inhibitors. PMID- 3038415 TI - Efficacy and influence on quality of life of enalapril as a first step treatment of hypertension. AB - 205 patients with mild to moderate uncomplicated hypertension participated in a six-month double-blind parallel study performed in a unique center. After a two week single-blind placebo period, the patients were randomly allocated to receive either Enalapril 20 mg once-a-day or placebo as the first step treatment. They were then followed-up every two weeks and successive doses of hydrochlorothiazide, oxprenolol, and dihydralazine were added until the diastolic pressure was lower than 90 mmHg. After six months, the systolic and diastolic blood pressures were lower in the Enalapril than in the control group (129/82 +/- 12/6 mmHg versus 136/86 +/- 10/5 mmHg; p less than 0.001). Drug withdrawal was necessary for 8 patients in the Enalapril group and for 16 patients in the control group (p less than 0.05). The number of daily tablets was 2.7 +/- 1.8 in the Enalapril group and 4.4 +/- 2.4 in the control group (p less than 0.01). Therefore, a stepped-care program based on Enalapril appears significantly more effective than a stepped-care program based on a diuretic. PMID- 3038416 TI - Clinical response to angiotensin-converting enzyme inhibition in cardiac failure. AB - The renin-angiotensin system plays a pivotal role in the pathophysiology of heart failure. ACE inhibitors block the production of angiotensin II and are efficacious in the therapy of patients with heart failure, ACE inhibitors have combined preload and afterload reducing effects. Cardiac index is augmented, cardiac filling pressures are reduced and renal function may improve with ACE inhibition. Long-term subjective and objective improvements have been reported. Preliminary data suggest that this class of vasodilators may favorably influence the prognosis of advanced heart failure. PMID- 3038417 TI - The renal response to converting enzyme inhibition and the treatment of sodium sensitive hypertension. AB - Multiple lines of evidence suggest that intrarenal angiotensin AII formation participates in the control of renal perfusion and function. Inappropriate activation of this intrarenal system may participate in the pathogenesis of hypertension in about 45 percent of patients with essential hypertension, a group that we call "non-modulators" (NM). In NM the renal vascular response to AII is blunted when the subjects are on a high-salt diet, but appropriate when they are on a low-salt diet. Converting enzyme inhibition in NM induces a larger increase in renal blood flow, than occurs in normal subjects or other patients with essential hypertension, most evident when they are on a high-salt diet. The renal vasodilatation is not due to prostaglandin or bradykinin accumulation in the kidney, since the increase in renal blood flow induced by converting enzyme inhibition is associated with an enhanced renal vascular response to AII. When NM are shifted from a low to a high sodium intake, they show more positive sodium balance, gain more weight and increase their blood pressure more--the characteristics of sodium sensitive hypertension. Converting enzyme inhibition corrects their inability to handle a sodium load as it improves renal blood flow, and induces a depressor response that does not correlate with plasma renin activity. Many of these characteristics are shown in a normotensive offspring of essential hypertensives: since sodium handling is genetically determined, this abnormality may represent the inherited renal abnormality. An abnormality in the control of the renal circulation by AII, reversed by converting enzyme inhibition, may represent a fundamental abnormality in the pathogenesis. PMID- 3038418 TI - Indomethacin reduces the antihypertensive action of enalapril. AB - We evaluated the influence of indomethacin on the pharmacological actions of Enalapril in 9 uncomplicated essential hypertensives. While on chronic treatment with Enalapril, these patients randomly received indomethacin (50 mg bid) or a corresponding placebo for 1 week and the opposite treatment after a 2 week interval. Indomethacin, which decreased serum thromboxane B2 and urinary 6-keto prostaglandin-F1 alpha, reduced the plasma renin activity (PRA) increased by Enalapril. Indomethacin did not modify serum ACE, whose activity had been reduced by the ACE inhibitor. Mean blood pressure (MBP) values, which were significantly and to a similar extent reduced by Enalapril at the beginning of the cross-over, after placebo addition and at the end of the two week interval, were significantly increased by indomethacin, despite being still significantly lower than baseline values. These data show that systemic and renal prostaglandin (PG) synthesis inhibition induced by indomethacin can blunt the antihypertensive effect of chronic Enalapril treatment in patients with essential hypertension. PMID- 3038419 TI - Role of endogenous angiotensin II and prostaglandins in the antihypertensive mechanism of angiotensin converting enzyme inhibitor in hypertension. AB - The role of endogenous angiotensin II and prostaglandins (PGs) in the antihypertensive effect of converting enzyme inhibitor, captopril, was studied in essential hypertension by the separate and the combined administration of captopril and indomethacin. Although plasma angiotensin II was similarly suppressed by the separate and the combined administration of captopril and indomethacin, captopril lowered and indomethacin increased the mean blood pressure. There were negative correlations between the changes in mean blood pressure and in urinary sodium excretion as well as in urinary PGE excretion. These results suggest that endogenous PGs may be implicated in the antihypertensive effect of captopril through the alteration of sodium balance. PMID- 3038420 TI - Effects of captopril on cardiorenal hemodynamics in patients with severe chronic congestive heart failure. AB - The effects of captopril on cardiorenal function were studied in patients with chronic congestive heart failure. A single oral dose of captopril in 16 patients significantly increased cardiac indices and decreased total systemic vascular resistance. These changes were greater in subjects with higher baseline plasma renin activity (PRA). The increase in PRA and decrease in plasma aldosterone were also greater in this group. During 7 days of captopril therapy, renal plasma flow distinctly increased in 10 patients in whom renal function was followed. The increase in renal blood flow was greater in subjects with higher PRA. Simultaneous infusion of aprotinin in eight of these subjects did not affect the captopril-induced increase in renal plasma flow: these responses were the same in both PRA subgroups. The results suggest that captopril reduces total systemic vascular resistance in patients with chronic congestive heart failure through the inhibition of the renin-angiotensin system and the preferential renal vasodilating effect of captopril seems exclusively to be the sole result of this inhibition, with the kallikrein-kinin system or kinin-mediated prostaglandins not playing a major role. PMID- 3038421 TI - Effect of prolonged angiotensin converting enzyme inhibition on ambulatory blood pressure of normotensive volunteers. AB - The blood pressure effect of 8-day angiotensin converting enzyme (ACE) inhibition was studied in normotensive healthy volunteers maintained on an unrestricted salt intake. Six volunteers received cilazapril (5 mg/day), 5 perindopril (8 mg/day) and 5 CGS14824A (10 mg/day). The 3 investigational inhibitors were found to have no consistent effect on blood pressure monitored during day-time outside the clinic using a semi-automatic blood pressure recorder (Remler M2000). PMID- 3038422 TI - Comparison of sublingual and oral captopril in hypertension. AB - The use of sublingual captopril has been recently suggested in hypertensive crisis on the assumption of a faster absorption and thus a more rapid effect on blood pressure than with the oral route. To verify this hypothesis we have compared the hypotensive effect of oral and sublingual captopril in 40 essential hypertensives who were randomly allocated to either route of administration. Captopril was administered orally dissolved in water or allowed to dissolve under the tongue. After 5, 10, 20, 30, 40, 60 and 90 minutes blood pressure, Plasma Renin Activity (PRA) and Angiotensin Converting Enzyme (ACE) were measured. No significant differences were found between the two groups in the time course of blood pressure decrease, PRA increase and ACE inhibition. The changes of the parameters studied was superimposable irrespective of the route of administration thus not supporting the hypothesis that sublingual captopril might be absorbed more rapidly. PMID- 3038423 TI - The effects of twice daily captopril and once daily enalapril on ambulatory intraarterial blood pressure in essential hypertension. AB - Intraarterial ambulatory pressure (AP) was recorded before and during therapy with captopril or enalapril in two groups with hypertension. Seven patients were admitted during the study. The monitoring of AP and heart rate (HR) was performed during placebo therapy and following a minimum period of 7 days of 25 mg twice daily captopril or 2.5 to 10 mg once daily enalapril. The AP and HR following percutaneous insertion of a cannula into the brachial artery were sampled then data were analyzed as reported previously. After the cannula was inserted, examinations of tilt-up, handgrip and ergometer were performed. Both drugs produced a significant reduction of ambulatory AP throughout 24 hours with preservation of the overall shape of the circadian curve. The results also demonstrated that both drugs had not affected normal daily activities. Thus, twice daily captopril and once daily enalapril can be used as the first-line therapy of hypertension. PMID- 3038425 TI - Single oral administration of captopril may not bring an improvement in screening of primary aldosteronism. AB - Plasma renin activity (PRA) and plasma aldosterone concentration (PAC) were measured in 19 cases with primary aldosteronism (PA) and 72 with essential hypertension (EHT) to differentiate the two disorders in the following conditions: after overnight recumbency (basal state) and after oral administration of captopril. Screening criteria were determined in order to pick up all of PA patients as positive. After the captopril administration, the specificity of a criterion based on the combination of PAC and PAC/PRA ratio was 93% and positive predictive value 79%. This criterion was superior to other conventional screening methods. However, higher specificity (97%) and positive predictive value (90%) were obtained from a combination criterion based on the basal PAC and PAC/PRA ratio. The single oral administration of captopril may not bring an improvement in the screening of PA. PMID- 3038424 TI - Role of renin-angiotensin and kallikrein-kinin systems on the mechanism of the hypotensive effects of converting enzyme inhibitor, alacepril. AB - In four patients with essential hypertension and one patient with renovascular hypertension, decreases in blood pressure and plasma angiotensin II levels, and increases in plasma renin activity and plasma kinin levels were observed during eight days of alacepril treatment. Significant correlations between the changes in mean arterial pressure and those in plasma angiotensin II or kinin levels were observed positively or negatively, respectively, in the essential hypertensives. These findings suggest that the hypotensive effect of alacepril might be caused mainly by a decrease in plasma angiotensin II levels and, at least in part, by an increase in plasma kinin levels. PMID- 3038426 TI - Long-term effects of converting enzyme inhibitors on split renal function in renovascular hypertension. AB - Long-term effects of angiotensin converting enzyme inhibitors on split renal function were studied using a noninvasive radioisotopic method in five patients with renovascular hypertension. In the stenotic side the glomerular filtration rate which acutely decreased following captopril, tended to return toward pretreatment levels on prolonged captopril administration (up to 2 years), while the value remained unchanged in the nonstenotic side of the kidney. Effective renal plasma flow increased in both sides. During long-term captopril treatment, there were no significant changes in serum creatinine, sodium, potassium, and blood pressure control was maintained at the same level in an acute period. These results suggest that the glomerular filtration in the stenotic kidney of renovascular hypertension, which was acutely reduced, will not further deteriorate, but will increase after prolonged captopril treatment. PMID- 3038427 TI - Side effects and metabolic effects of converting enzyme inhibitors. AB - Captopril Research Group of Japan made an evaluation of clinical usefulness of captopril in the treatment of mild to moderate essential hypertension. A relatively low frequency of side effects manifested as clinical symptoms was observed when the daily dose of captopril used was 37.5-112.5 mg. Antihypertensive effect was well maintained in this dose range. Follow-up surveillance of captopril also confirmed this. Abnormalities in blood chemistry were also found to be very less frequent. Untoward metabolic effects of ACE inhibitors such as alteration of serum electrolytes, lipid profiles or glucose metabolism were reported to be minimal or not actually observed. Side effects and untoward metabolic effects may appear more frequently when the same dose was given in patients with renal impairment. ACE inhibitors can be used as the first choice of drugs in the treatment of essential hypertension with no renal function impairment. PMID- 3038428 TI - Metabolic and blood pressure effects of angiotensin converting enzyme inhibitors in mild-to-moderate American hypertensives. AB - To evaluate the effects of angiotensin converting enzyme (ACE) inhibition on metabolic effects as well as blood pressure, we studied 255 mild-to-moderate American essential hypertensives. After a 4-week placebo period subjects were randomly assigned to receive captopril (C) 25 mg tid, hydrochlorothiazide (H) 15 mg tid or both agents in combination (C j) tid. A significant (p less than 0.05) fall in blood pressure was seen in all three groups; C + H had a greater (p less than 0.05) than C or H. H was associated with a significant (p less than 0.05) fall in serum potassium and significant (p less than 0.05) increases in uric acid, glucose and cholesterol. With C alone no changes in these parameters were seen. With combination therapy (C + H) significant (p less than 0.05) blunting of diuretic-induced changes in potassium and uric acid and the significant increases in glucose and cholesterol seen with H were prevented. The side effects reported were mild with all regimens and rarely required discontinuation of therapy. The efficacy, safety and advantages of ACE inhibition make it attractive as primary treatment in mild-to-moderate hypertension. PMID- 3038429 TI - Side effects and metabolic effects of converting-enzyme inhibitors. AB - Side effects of angiotensin converting enzyme (ACE) inhibitors are not common with currently recommended doses. Hypotension, hyperkalemia and renal impairment may occur under special circumstances, and relate directly to blockade of ACE, in particular when pre-treatment renin levels are high. Other adverse effects, once quite common when excessive doses of captopril were prescribed, are now rarely seen. A possible exception is cough. Though not dangerous, cough is not infrequently seen during treatment with ACE inhibitors. Experience with these drugs is insufficient to recommend their use in the hypertension of pregnancy. The "metabolic-profile" with ACE inhibitors appears favourable, since they can increase uric acid excretion and lower plasma urate levels, carbohydrate tolerance is unaltered or improved, and lipid levels are unchanged. Electrolyte and metabolic effects of thiazide diuretics are blunted by addition of an ACE inhibitor. PMID- 3038430 TI - Angiotensin converting enzyme inhibition reduces ACTH release due to hypoglycaemia. AB - To investigate the role of Angiotensin II in the release of ACTH, the response of adrenocorticotrophic hormone to hypoglycaemia was studied before and during treatment with an angiotensin converting enzyme inhibitor, enalapril, in 15 male patients with essential hypertension. Plasma levels of ACTH were measured before and 60, 90 and 120 min after an i.v. bolus of normal saline, as placebo, and, 3 days later, after an i.v. bolus of regular insulin (0.15 U/Kg b.w.). Enalapril treatment was then started and both placebo and hypoglycaemic tests were repeated 15 days thereafter. No changes in ACTH plasma levels were observed after acute normal saline either before or during enalapril treatment. On the contrary, hypoglycaemia induced a sharp increase of ACTH before enalapril (from 19.5 +/- 4.1 to 74.4 +/- 13.0 pg/ml, p less than 0.01 60 min after insulin) but not during ACE inhibition (from 26.1 +/- 6.2 to 34.6 +/- 5.9 pg/ml, NS, at min 60 of the study). The present data confirm our previous observation on the reduction of the hypoglycaemic-induced ACTH release during ACE inhibition with captopril and support the hypothesis that circulating Ang II may exert a facilitating role on adrenocorticotrophic hormone release. PMID- 3038431 TI - The metabolic effects of enalapril. AB - The metabolic effects of Enalapril (EN) were compared in a cross over study with those of Propranolol (PPL). Long term treatment with the ACEI did not modify the lipid and glucose profile and increased urate excretion, while PPL increased triglycerides and decreased urate clearance. PMID- 3038432 TI - Effect of chronic treatment with angiotensin I converting enzyme inhibitors on circulating atrial natriuretic polypeptide in spontaneously hypertensive rats. AB - We have recently shown that circulating atrial natriuretic polypeptide (ANP) in adult spontaneously hypertensive rats (SHR) is higher than that in age-matched Wistar-Kyoto rats (WKY). The present experiment was designed to examine the possible effects of chronic treatment with angiotensin I converting enzyme inhibitors (ACEI) on plasma ANP levels in SHR. Captopril and enalapril lowered blood pressure and reduced relative ventricular weight in SHR but not to the level of WKY. Plasma ANP levels were decreased by captopril and enalapril compared with untreated SHR. These results suggest that the ANP release may be suppressed in ACEI-treated SHR compared with untreated SHR. We speculate that a reduction of cardiac overload by ACEI may in part explain the decline of circulating ANP. PMID- 3038433 TI - A combined approach to the assessment of neurological dysphagia. AB - The dysphagia encountered by patients with neurological disorders can be both distressing and life threatening because of the associated problems of aspiration. Decisions regarding management are often difficult because the exact nature of the underlying disorder varies from patient to patient and is frequently complex. A new approach to the assessment and evaluation of acquired neurological dysphagia is presented. Management of the first 50 patients assessed by this method is described. The advantages of a joint dysphagia clinic comprising neurology, speech therapy, ENT and radiology departments are discussed. PMID- 3038434 TI - Bile acids and cancer of the large bowel. PMID- 3038435 TI - Current perspectives on hepatocellular carcinoma in Oriental and African countries compared to developed western countries. PMID- 3038436 TI - Decreased interleukin 1 activity in culture supernatant of lipopolysaccharide stimulated monocytes from patients with liver cirrhosis and hepatocellular carcinoma. AB - Immunoregulatory function of peripheral blood monocytes was studied in patients with hepatocellular carcinoma (HCC) and liver cirrhosis (LC), by assaying interleukin 1 (IL-1) and prostaglandin E2 (PGE2) in the culture supernatant of lipopolysaccharide-stimulated monocytes. IL-1 activity of the monocyte culture supernatant without indomethacin was decreased in patients with HCC and LC, compared with that of controls. The activity was lower in patients with HCC than that in those with LC. The PGE2 content of the culture supernatant of monocytes from patients with LC and HCC was increased, compared to normal controls. To avoid the effect of PGE2 on the IL-1 assay, we cultured the monocytes with addition of indomethacin and assayed IL-1 activity in the culture supernatant. As a result, monocyte IL-1 production was increased in patients with HCC and LC, compared with normal controls. The decrease in IL-1 activity of the supernatant without indomethacin of patients with LC and HCC was considered to be due to increased secretion of PGE2 by the monocytes. Therefore, monocytes from patients with HCC and LC had an increased capacity of secreting both IL-1 and PGE2 over normal controls, but the effect of the suppressor function (PGE2 secretion) dominated in these patients. PMID- 3038437 TI - Immunohistological evidence of lymphokine production and lymphocyte activation antigens in tuberculin reactions. AB - Evidence of lymphokine elaboration and lymphocyte activation was sought in tuberculin skin test reactions at 24 and 48, or 48 and 96 h in patients with active, culture-proven, pulmonary tuberculosis. Through the use of frozen sections, immunoperoxidase techniques and monoclonal antibodies, anti-interleukin 2 positive cells were found to constitute 0.4% to 0.6% of the dermal infiltrate, and keratinocyte Ia expression at 96 h was consistent with a marker for interferon-gamma production. Cells bearing the interleukin 2 receptor more than doubled in prevalence from 24 to 48 or 96 h but cells staining with Ta1, an antibody identifying activated lymphocytes, were 10% of the cells of the infiltrate at all three times. One-half of the cells of the infiltrate were OKM1 positive, presumably macrophages, perhaps reflecting the presence of active tuberculosis. PMID- 3038439 TI - Increased half-life of gammaglobulin after prolonged intravenous replacement therapy. AB - The half-life of total IgG and antibodies to cytomegalovirus and pneumococcal polysaccharide type III was consistently prolonged after a 9 month course of intravenous gammaglobulin treatment in three patients with 'primary' hypogammaglobulinaemia. The half-life of antibody to tetanus toxoid showed no consistent pattern. These data are at variance with the traditional dogma that IgG half-life is inversely related to serum IgG concentration. They suggest that patients on intravenous gammaglobulin therapy are likely to need less, rather than more, infusions when their serum IgG has risen to within the physiological range. PMID- 3038438 TI - Modification of beta-2-microglobulin in sera from patients with small cell lung cancer: evidence for involvement of a serine protease. AB - Modification of beta-2-microglobulin has been shown to occur in vitro in serum of patients suffering of small cell lung cancer, where it clearly correlated to the clinical course of disease. The cause of serum beta-2-microglobulin modification occurring in these patients is investigated in the present study. This revealed that modification of beta-2-microglobulin is due to cleavage by a serine protease, which is dependent on divalent cations and has a trypsin-like specificity. The process showed the following characteristics: pH optimum at 8.5, heat inactivation by incubation at 56 degrees C for 20 min and temperature optimum at 20 degrees C. PMID- 3038440 TI - Antibody-independent activation of the classical pathway of complement by Epstein Barr virus. AB - A purified preparation of Epstein-Barr virus (EBV) has been shown to activate the classical complement pathway by direct interaction with the first component of complement, C1, without the intervention of antibody. No evidence was found for activation of the alternative pathway. Following classical pathway activation the specific affinity of EBV for B cells can be presumed to be lost since the virus will become opsonized for clearance by phagocytic cells bearing complement receptors, CR1 and CR3. This activation is further evidence that complement plays a role in defence mechanisms independently of antibody activity. PMID- 3038441 TI - IL-2 receptor and HLA class II antigens on cerebrospinal fluid cells of patients with multiple sclerosis and other neurological diseases. AB - The presence of IL-2 receptor and HLA class II antigens as detected by monoclonal antibodies on mononuclear cells from both cerebro-spinal fluid (CSF) and peripheral blood was examined by cytofluorographic analysis in patients with multiple sclerosis (MS) and other neurological diseases. CSF as compared to blood was enriched in cells expressing IL-2 receptor and HLA class II molecules both in MS patients and in other inflammatory diseases of the central nervous system suggesting that activated T-cells concentrate within the central nervous system. PMID- 3038443 TI - Activation by puff adder venom of inactive renin in normal and hypertensive rat plasma. AB - Inactive renin has been studied extensively in human plasma, but in animal plasma its accurate quantification has proved more difficult, due partly to higher activity of plasma protease inhibitors. Such activity in human plasma can be conveniently destroyed by a metalloprotease in Bitis arietans venom, with concommitant release of endogenous enzyme activities, such as plasma kallikrein, that then activate inactive renin. It was therefore of interest to look for inactive renin in rat and rabbit plasma using this approach, so providing, in addition, a comparison for the disparate data of other groups who have used trypsin or acid for activation. In both rat and rabbit plasma the proportion of inactive renin was 62% of total renin, whereas human plasma contained more inactive renin and a higher proportion, 82%. A higher concentration of venom was required for rat (33 ug venom/ml plasma) and rabbit (4 micrograms/ml) than needed for activation, at a similar rate, in human plasma (1 microgram/ml). When applied to studies of rats made hypertensive and hyper-reninaemic by aortic ligation for 5 days, higher total (active + inactive) renin was observed. The proportion of inactive renin, as a percentage of total renin in plasma collected at this time, was, however, found to diminish significantly. In conclusion, puff adder venom activates inactive renin in rat and rabbit plasma and can be used to study physiological changes in inactive renin in such animal plasma. PMID- 3038442 TI - The role of reactive oxygen intermediates in nonspecific monocyte cytotoxicity induced by immune complexes. AB - Normal human monocytes were induced to lyse nonsensitized target cells when triggered by precipitating immune complexes (IC) or soluble heat-aggregated IgG (HAIgG). Catalase, azide, cyanide and three aminoacids employed as quenchers of ClO, significantly inhibited this nonspecific cytotoxicity (NSC), suggesting an important role for the myeloperoxidase (MPO) system. However, HO and/or 1O2 may also be involved in the lysis, since certain scavengers of these species such as mannitol, benzoate, ethanol and histidine, as well as superoxide dismutase (SOD), partially inhibited NSC. Moreover, cyanide and azide were unable to completely abrogate this lytic activity. When NSC was compared to antibody dependent cellular cytotoxicity (ADCC), it was found that neither catalase nor oxygen species scavengers affected ADCC while azide and cyanide significantly enhanced it. Antibody-coated target cells were also destroyed by IC-triggered monocytes. However, kinetic analysis and studies on the capacity of catalase to inhibit the lysis demonstrated that it was mediated through a NSC-like mechanism. The cytotoxic system described in this report offers a suitable model to study in vitro alternative lytic mechanisms triggered through monocyte receptors for the Fc portion of IgG (Fc gamma R). PMID- 3038444 TI - Dose-response relationships for adrenocorticotrophin-induced hypertension in man. AB - Adrenocorticotrophin (ACTH), given as long-acting Synacthen depot (Ciba-Geigy), at 1000 micrograms/day, in divided doses 12 hourly, is known to increase blood pressure in man. Fifty micrograms/day of short-acting Synacthen given intravenously produced a rise in blood pressure and may be a threshold dose. Twelve hourly intramuscular injections of short-acting Synacthen over the dose range 100-400 micrograms/day was not sufficient to raise blood pressure. PMID- 3038445 TI - Infectious causes of fetal death. PMID- 3038446 TI - Fixation of experimental osteotomy of the distal femur with biodegradable thread in rabbits. AB - A distal femoral osteotomy on 24 rabbits was fixed with biodegradable polyglycolic acid (PGA) thread pulled through drill holes made on both sides of the osteotomy. Follow-up times were one, three, six, 12 and 24 weeks. The distal part of each femur was removed, fixed in alcohol, embedded in methylmethacrylate, sawed to 80 microns thick for oxytetracycline (OTC)-labeling studies and microradiography, and sectioned at 5 microns for histologic studies. As judged by histologic microradiographic, and OTC-labeling studies, 19 of 24 osteotomies were healing normally; after six weeks of follow-up examination, union of 11 of 14 osteotomies was observed on radiographs. On the basis of the present study, PGA threads may be promising for the fixation of osteotomies of the metaphyseal cancellous bone in rabbits. PMID- 3038448 TI - Resolution of a photopenic lesion on bone imaging following chemotherapy for metastatic disease. AB - Resolution of abnormal foci of increased radiotracer deposition on bone imaging occurs commonly. Photopenic areas due to avascular necrosis and infarcts have been reported to resolve. This report describes serial changes in a photopenic lesion due to metastatic disease and demonstrates that such lesions can resolve following chemotherapy. PMID- 3038447 TI - Myocardial infarct imaging in patients with technetium-99m 2,3-dimercaptosuccinic acid. Superiority of technetium-99m pyrophosphate. AB - Technetium-99m 2,3-dimercaptosuccinic acid (Tc-99m DMSA) has been used successfully for imaging acute myocardial infarction in a canine model. The application in humans, however, has not been previously reported. In order to determine the feasibility of using this agent in clinical studies and to compare the agent to technetium-99m pyrophosphate (Tc-99m PPi), ten patients with proven myocardial infarction were studied. While imaging of transmural infarctions in humans was achieved using Tc-99m DMSA, scores for the Tc-99m DMSA images (1.8 +/- 0.96) were not as high as for Tc-99m PPi (2.5 +/- 0.45) (P less than 0.05). Discordance among four independent interpreters was greater for images obtained with Tc-99m DMSA. The superiority of Tc-99m PPi was evident whether images were obtained early (within 24 hours) or late (within five days). Although DMSA images were not obscured by rib uptake, they were less sensitive (63%) than Tc-99m PPi (97%). A potential advantage of Tc-99m DMSA in imaging acute myocardial infarction is that radiotracer concentration in the infarct occurs primarily in the early postinfarction period. The longer postinfarction that Tc-99m DMSA imaging was attempted, the lower the concentration of radiotracer. Thus, Tc-99m DMSA would not be expected to have the same persistence pattern as Tc-99m PPi into the remote postinfarction period. The persistent positivity of Tc-99m PPi has made it difficult to diagnose reinfarction. PMID- 3038449 TI - Scrotal scan in traumatic hematoma. PMID- 3038450 TI - Radionuclide imaging of primary tumors and tumor-like conditions of the liver. AB - The unique information provided by radionuclide techniques allows the noninvasive characterization of primary tumors and tumor-like conditions of the liver. A thorough knowledge of the mechanisms of radiopharmaceutical localization and the histologic abnormalities present in each condition is required to optimally use these techniques. A review of this subject is presented. PMID- 3038451 TI - Effect of structured dietary fiber on bioavailability of amoxicillin. AB - The effect of structured dietary fiber on the bioavailability of amoxicillin (AMX) was evaluated. Ten healthy volunteers ingested one of two isocaloric, isonitrogenous diets providing 7.8 gm/day (diet I) and 36.2 gm/day (diet II) of structured fiber for 3 days. Then they ingested one tablet (500 mg) AMX after breakfast. The other diet was administered for an additional 3 days and the study was repeated. Plasma and urinary AMX concentrations were measured at 9 and 24 hours, respectively, by a microbiologic technique. An open one-compartment model was used for pharmacokinetic analysis. AMX was absorbed more slowly when ingested with diet I than with diet II: the absorption rate constant was 1.04 +/- 0.37 and 1.75 +/- 0.75 (P less than 0.05); lag time for absorption was 0.34 +/- 0.13 hours and 0.29 +/- 0.11 hours (P less than 0.05). The first-order rate constant and elimination half-life were similar. Bioavailability was higher with diet I: the AUC was 12.17 +/- 3.04 vs. 9.65 +/- 2.64 micrograms/ml/hr with diet II (P less than 0.05). A higher content of dietary fiber increased AMX absorption rate and decreased the amount of drug absorbed. PMID- 3038452 TI - Levels of serum angiotensin-converting enzyme before and after forearm venous stasis in diabetic microangiopathy. AB - The levels of angiotensin-converting enzyme and Factor VIII-related antigen, 2 endothelial synthesized glycoproteins, were measured in basal conditions and after forearm venous stasis in 12 healthy controls and 12 patients with diabetic microangiopathy. Angiotensin-converting enzyme levels were similar in the controls and in the patients both basally (185 +/- 17 nm/ml/min (SEM) and 192 +/- 15, respectively) and after stasis (238 +/- 17 and 220 +/- 15), whereas Factor VIII-related antigen was lower in the former basally (101 +/- 13% vs 184 +/- 16%, p less than 0.001) and after stasis (140 +/- 21% and 243 +/- 21%, p less than 0.01). It is concluded that Factor VIII-related antigen is a more sensitive indicator of endothelial cell damage in diabetic microangiopathy than angiotensin converting enzyme. PMID- 3038453 TI - Different effect of two immunomodulating agents cyclosporin A and ciamexon on the insulin metabolism of cultured mouse islets. AB - Ciamexon is a new immunomodulating agent with promising effects in treatment of newly-diagnosed insulin dependent diabetic patients. Ciamexon seems to have fewer adverse toxic properties than Cyclosporin A. Therefore, the effects of both substances on islet cell function were studied. Collagenase-isolated mouse islets were cultured free-floating for 3 or 7 days in medium RPMI 1640 plus 10% fetal calf serum in the presence of Cyclosporin A or Ciamexon (0.1 microgram/ml, 1 microgram/ml). Culture of the islets in the presence of 0.1 microgram/ml Ciamexon neither reduced the number of viable islets nor impaired insulin secretion as was found in Cyclosporin A. These results should be taken into account when treating early Type 1 diabetes or when transplanting islet cells. PMID- 3038455 TI - Erythrocyte phosphoinositide metabolism in essential hypertensive patients and their normotensive offspring. AB - The metabolism of phosphoinositides was investigated in erythrocyte membranes of essential hypertensive patients, normotensive offspring of hypertensive patients and matched controls. Measurement of 32P-labelling of phosphatidylinositol 4 phosphate and phosphatidylinositol 4,5-bisphosphate revealed no differences in the rate of incorporation of the isotope in essential hypertensive patients compared with controls. In the normotensive offspring of essential hypertensive patients there was a highly significant increase in the rate of 32P incorporation (P less than 0.01), compared with matched controls, indicating a higher rate of metabolic turnover in these subjects. These data demonstrate that phosphoinositide metabolism is enhanced in subjects genetically at risk of hypertension, before the blood pressure has risen, but once the blood pressure is established, it is no different from control values. Abnormal phosphoinositide metabolism may be a further manifestation of a genetically determined defect of membrane physicochemical function underlying essential hypertension. PMID- 3038454 TI - Dietary supplementation of omega-3 polyunsaturated fatty acids improves insulin sensitivity in non-insulin-dependent diabetes. AB - The effect of dietary omega-3 polyunsaturated fatty acids on the lipid composition and fluidity of erythrocyte membranes, and on in vivo insulin sensitivity was studied in 6 non-insulin-dependent diabetic (NIDD) patients. An 8 weeks daily supplementation of 3 g of the omega 3 fatty acids eicosapentaenoic and docosahexaenoic acid resulted in an increase of the membrane phospholipid unsaturation and the sphingomyelin content. Erythrocyte membrane fluidity, measured with electron spin resonance of intact erythrocytes and with fluorescence polarization of erythrocyte ghosts, did not change. The in vivo insulin stimulated glucose uptake was estimated by determining the metabolic clearance rate (MCR) of glucose in the steady state of a simultaneous infusion during 150 min of glucose (33 mumol/kg/min) and insulin (50 mU/kg/hr). The MCR of glucose increased in all patients; from 3.93 +/- 0.55 - 4.69 +/- 0.74 ml/kg/min (mean +/- SEM, p less than 0.05). Plasma triglyceride concentrations fell from 1.9 +/- 0.3 - 1.2 +/- 0.2 mmol/l (mean +/- SEM, p less than 0.05). We conclude that in NIDDs dietary supplementation of omega 3 fatty acids improves in vivo insulin sensitivity and lowers plasma triglyceride levels, while erythrocyte membrane fluidity remains unaltered. PMID- 3038456 TI - Water depletion, not oral sodium loading, increases levels of sodium, potassium dependent adenosine triphosphatase inhibitors in rat plasma. AB - In order to define a physiological role for circulating inhibitors of sodium, potassium-dependent adenosine triphosphatase (Na+,K+-ATPase), plasma was obtained from control, water deplete, water repleted, sodium deplete and sodium loaded rats. The effect of this plasma on Na+,K+-ATPase activity, and its transport equivalent 86Rb uptake, was measured in separated guinea pig renal cortical tubules. Plasma from water deplete rats had a raised plasma osmolality and sodium concentration and a significant inhibitory effect on Na+,K+-ATPase (14%) and 86Rb uptake (24%) compared with control or water repleted rats. Inhibition of Na+,K+ ATPase and 86Rb transport was not seen with plasma from rats after dietary sodium loading (urine sodium 5.2 +/- 0.9 mmol/day) compared with low sodium diet controls (urine sodium 0.41 +/- 0.08 mmol/day). Des-amino arginine vasopressin in vivo produced no inhibition of Na+,K+-ATPase or Rb transport. These studies suggest, that in terms of common homoeostatic insults, circulating inhibitors of Na+,K+-ATPase are more responsive to water depletion than to oral sodium loading. The inhibitors may fulfil a physiological role in increasing sodium excretion to maintain osmolality after dehydration. PMID- 3038457 TI - Proteinases and eukaryotic cell growth. PMID- 3038458 TI - Changes in the viscosity of the plasma membrane of flounder (Platichthys flesus L.) erythrocyte induced by varying the content of membrane cholesterol or by benzyl alcohol. Correlation of the activity of the intrinsic Mg2+-ATPase and the viscosity. AB - The order parameter S and the correlation time tau from the ESR-spectra of 5-DS and 5-DD, respectively, are used as measures of the viscosity of the plasma membrane. It is suggested that the membrane undergoes a thermotropic phase transition which correlates with a change of the activation energy of the (Na+ + K+)-ATPase but not of the Mg2+-ATPase. The viscosity varies almost proportionally with the amount of cholesterol in the membrane when 0 less than mg C/mg PL less than 0.5. It is suggested that cholesterol is uniformly distributed in the membrane when mg C/mg PL less than 0.5 and that cholesterol-rich domains appear when mg C/mg PL greater than 0.5. Benzyl alcohol decreases the viscosity of the membrane. The effect depends on the presence of cholesterol in the membrane. The onset of the effect of benzyl alcohol on the native membrane is delayed--in terms of alcohol concentration--relative to the cholesterol-depleted membrane. Benzyl alcohol affects only the outer part of the membrane bilayer. Changes of the viscosity and of the activity of the Mg2+-ATPase are proportional. The effect of cholesterol-depletion on the activity of the Mg2+-ATPase cannot be ascribed only to the simultaneous lowering of the viscosity. PMID- 3038459 TI - Metabolism of glycerate 2,3-P2--XII. Characterization of the 2,3 bisphosphoglycerate synthase-phosphatase and of the hybrid phosphoglycerate mutase/2,3-bisphosphoglycerate synthase-phosphatase from pig brain. AB - 2,3-Bisphosphoglycerate synthase-phosphatase and the hybrid phosphoglycerate mutase/2,3-bisphosphoglycerate synthase-phosphatase have been partially purified from pig brain. Their 2,3-bisphosphoglycerate synthase, 2,3-bisphosphoglycerate phosphatase and phosphoglycerate mutase activities are concurrently lost upon heating and treatment with reagents specific for histidyl, arginyl and lysyl residues. The two enzymes differ in their thermal stability and sensitivity to tetrathionate. Substrates and cofactors protect against inactivation, the protective effects varying with the modifying reagent. The synthase activity of both enzymes shows a nonhyperbolic pattern which fits to a second degree polynomial. The Km, Ki and optimum pH values are similar to those of the 2,3 bisphosphoglycerate synthase-phosphatase from erythrocytes and the hybrid enzyme from skeletal muscle. The synthase activity is inhibited by inorganic phosphate and it is stimulated by glycolyate 2-P. PMID- 3038460 TI - Structure of the 3' region of the human elastin gene: great abundance of Alu repetitive sequences and few coding sequences. AB - Two overlapping clones encompassing 8.5 kb of the human elastin gene were isolated from two genomic libraries constructed by partial digestion with either HaeIII/AluI or Sau3A and contained in lambda Charon 4A or EMBL3, respectively. The 6 kb of DNA comprising the most 3' portion of the gene were sequenced demonstrating an extremely low coding ratio since only three exons containing a total of 134 translated nucleotides were identified. Two exons totaling 78 bp of translated sequences which were previously found in the bovine gene were absent in the human gene. The 3' most exon encoded the unusual amino acid sequence, GGACLGKACGRKRK. The human gene was terminated by 1.2 kb of untranslated sequence which contained two polyadenylation attachment signals. The remainder of the 6 kb was composed of intervening sequences which were abundantly rich in Alu family repetitive sequences found in both orientations. This first report of the characterization of the human elastin gene suggests that significant variation in the gene may exist between species and raises the possibility of consequential polymorphism, mediated by recombination between Alu sequences, in the human population. PMID- 3038461 TI - Captopril challenge with radionuclide assessment in two-kidney and one-kidney Goldblatt hypertension. PMID- 3038462 TI - Interactive functional imaging of renal scintigraphy after captopril in the detection of global and segmental hypoperfusion. PMID- 3038463 TI - Nephrotoxicity of ciclosporin in transplanted kidneys demonstrated with the 99mTc perfusion studies. PMID- 3038464 TI - Thallium-201 and technetium-99m subtraction scanning of the parathyroid glands in patients with hyperparathyroidism due to renal osteodystrophy. PMID- 3038465 TI - Assessment of renal function and scarring: is a DMSA scan always necessary? PMID- 3038466 TI - The current status of renal radiopharmaceuticals. A review. PMID- 3038467 TI - Renal functional changes after converting enzyme inhibition or nitroprusside in hypertensive rats. PMID- 3038468 TI - DNA hybridization of cervical tissues. AB - The increasing frequency of cervical neoplasia among younger women and the increased invasiveness of these tumors has led to a considerable growth in research into this disease. Conventional methods (epidemiology, cytology, and immunology), while being extremely useful, also have significant limitations. Recent advances in techniques for the manipulation of DNA now make it possible to analyze tissues for the presence of viral genomes. This review introduces these techniques and describes their application to the search for herpes simplex virus and human papillomavirus sequences in cervical tissue. The significance of the findings both for the mechanism of transmission of the disease, and also the consequences for early detection and hence more successful treatment, are also discussed. PMID- 3038469 TI - Epstein-Barr virus-associated malignancies. AB - The infectious aspects of cancer in humans were epidemiologically pioneered by Dr. David Burkitt through his observations of lymphomatous tumors seen in children in equatorial Africa. Years, later, the Epstein-Barr virus (EBV) was shown to be intimately associated with such tumors and is now recognized as a component of some B-cell lymphomas and nasopharyngeal carcinoma. Still the questions of an active, passive, or accessory role persist. The ability of this virus to cause immunosuppressive hemopoietic disturbances in individuals infected with EBV but not developing cancer raise questions about host susceptibility, host immune response, and possible coconspiring, infectious, oncogenic agents. Recent associations of EBV antibody found in diseases, such as squamous cell carcinoma of the head and neck and acquired immunodeficiency syndromes, point to its possible accessory role as an immunosuppressive agent. The ability of EBV to spread by extracellular and intracellular mechanisms demonstrates its variable infectious potential. Numerous EBV-transformed human cell lines attest to its ability to confer "immortality" with uncontrolled growth patterns. This review critically examines the association of EBV with various malignancies, the type of evidence which links it there, and the implications for further investigations and therapy. PMID- 3038470 TI - Pulmonary sulcus metastases simulating intraabdominal malignancy in childhood tumors. AB - Wilms tumor, the most common solid childhood malignancy, is frequently associated with pulmonary metastases, while hepatic metastases occur less frequently. Metastases to the lower lobes of the lungs, when deep in the costophrenic sulcus, may simulate an intraabdominal mass. The differentiation of these lesions is important both diagnostically and therapeutically. Three cases are presented in whom pulmonary sulcus metastases simulated abdominal lesions on computed tomography and could not be clearly localized as thoracic in origin on liver/spleen scintigraphy. Ultrasound evaluation was the most useful; however, integration with other imaging techniques was necessary to correctly identify these lesions as being of pulmonary origin. PMID- 3038471 TI - Cavitating metastases of the stomach and duodenum. AB - Three patients with cavitating gastric or duodenal metastases are presented. Metastatic disease, either by direct extension or by hematogenous spread, should be included in the differential diagnosis of an excavated exoenteric gastric or duodenal mass. PMID- 3038472 TI - Hepatolithiasis complicated by cholangiocarcinoma. AB - A case of hepatolithiasis complicated by intrahepatic cholangiocarcinoma is reported. Awareness of this association is important for the radiologist, who may detect clinically occult malignancy in patients with intrahepatic biliary calculi. PMID- 3038473 TI - Alpha-2 adrenergic modulation of norepinephrine secretion in the perfused rabbit iris-ciliary body. AB - Clonidine and other selective alpha-2 adrenergic agonists have been found to lower intraocular pressure in the eyes of rabbits and primates, including humans. It has been suggested that the ocular hypotensive response to alpha-2 agonists may be mediated, in part, by prejunctional inhibition of norepinephrine secretion at intraocular synapses. In this study, we have investigated the effects of adrenergic agonists and antagonists on field-stimulated, Ca++-dependent release of 3H-norepinephrine (3H-NE) from isolated, perfused rabbit iris-ciliary bodies and have utilized radioligand binding methods to identify prejunctional adrenoceptors in this tissue. Clonidine (10(-9)-10(-5) M) produced a dosage dependent inhibition of stimulation-evoked 3H-NE secretion (EC50 approximately equal to 3 X 10(-8) M), but did not alter basal secretion. Other adrenergic agonists capable of activating alpha-2 adrenoceptors (e.g., epinephrine, norepinephrine and xylazine) also significantly depressed 3H-NE secretion, whereas selective alpha-1 adrenergic or beta adrenergic agonists were without effect. Clonidine-mediated inhibition of 3H-NE release was reversed by the selective alpha-2 antagonist yohimbine (10(-7) M), but was unaffected by prazosin or timolol. Yohimbine alone markedly enhanced 3H-NE secretion, indicating tonic activation of prejunctional alpha-2 adrenoceptors by endogenous released norepinephrine. Forskolin or 8-bromo-cAMP, which alone enhanced norepinephrine secretion, failed to attenuate the inhibitory responses to alpha-2 agonists. 3H rauwolscine binding measurements showed a small decrease in alpha-2 receptor sites in iris-ciliary body membranes following surgical sympathetic denervation. It is concluded that the rabbit iris-ciliary body contains functional, prejunctional alpha-2 adrenoceptors which may play an autoregulatory role in vivo and contribute to the ocular effects of adrenergic drugs. PMID- 3038474 TI - Cytomegalovirus pneumonitis in patients with AIDS. Findings in an autopsy series. AB - Cytomegalovirus (CMV) pneumonitis is a common complication of AIDS. With the development of potentially effective drug therapy, the indications for treatment and best means of diagnosis have become issues of practical importance. We reviewed the clinical records and autopsy material from 54 patients dying with AIDS, 39 (72 percent) of whom had CMV infection documented postmortem by culture and the presence of inclusion bodies. Of the patients with CMV infection, disseminated disease was documented in 23 (59 percent) and pneumonitis in 31 (80 percent). Although the majority of patients with CMV pneumonitis had additional forms of pulmonary pathology, CMV was the only causative agent identified in two patients with severe pulmonary disease. Since CMV can cause severe lung damage and disseminated infection in patients with AIDS, specific treatment may be beneficial. Improved diagnostic technology is needed in this area. PMID- 3038475 TI - Postchemotherapy resection of residual tumor in limited stage small cell lung cancer. AB - To determine the feasibility of post-chemotherapy resection of residual tumor in small cell lung cancer, 24 selected patients with limited-stage disease were evaluated for exploratory thoracotomy. All 24 patients achieved partial or complete clinical response to chemotherapy and were considered adequate medical candidates for surgical resection. Fifteen patients underwent a lobectomy or pneumonectomy, 13 of whom had residual tumor in the resected specimen. Of the nine remaining patients, seven had no tumor found on biopsy at thoracotomy and two had unresectable mediastinal node involvement. No chemotherapy was administered postoperatively in any patient until disease progression or relapse was documented. Median survival for the entire group was 19 months and did not differ according to the surgical procedure performed. Nineteen patients had relapse. Patients undergoing biopsy only recurred locally in six of seven cases a median of five months post-thoracotomy (range one to six months). Two "biopsy only" patients were tumor free at 34+ and 56+ months. Local recurrence was observed in six of 12 resected patients, while six patients experienced only extrathoracic metastases. Median time to recurrence for resected patients was also five months. Four resected patients experienced late recurrence 16 to 36 months postoperatively and were alive with tumor at 29+ to 42+ months. Two resected patients were tumor-free at 13+ and 37+ months. Post-chemotherapy surgical resection was feasible in limited-stage patients and improved local control of disease. PMID- 3038476 TI - Silica-dust-exposed mine workers with scleroderma (systemic sclerosis). AB - The incidence of scleroderma (systemic sclerosis) was found to be increased in a population of black men who were gold miners. Ten men with scleroderma were detected during a five-year period. The annual incidence of the disease in this population in the group aged 33 to 57 years was estimated to be 81.8 per million. All of the men with scleroderma had disturbances of pulmonary function which were not present in a control group of silica-dust-exposed men without scleroderma. Not all of the subjects with scleroderma had silicosis, but all had been occupationally exposed to silica dust. There was a significant increase in the prevalence of tuberculosis in the past in the group with scleroderma, compared with a group of men with silicosis from the same population. The nature of the association of tuberculosis with scleroderma has not been defined. PMID- 3038477 TI - Spectrum of serum cortisol response to ACTH in ICU patients. Correlation with degree of illness and mortality. AB - Recent reports suggest adrenal insufficiency in critically ill patients is common. We found only one case of de novo adrenal insufficiency using admission ACTH injection in 70 selected intensive care unit (ICU) patients. Random serum cortisol levels correlated positively with illness severity in ICU patients using proven methods for assessing illness severity. Those with the highest random serum cortisol levels (greater than 60 micrograms/dl) had the greatest mortality, while those with lower random cortisol levels which stimulated to more than 18 micrograms/dl after ACTH injection had improved outcomes. Based on our results, routine screening for adrenal insufficiency in ICU patients is not warranted. If it is suspected, the cosyntropin test should be performed since low random cortisol levels (even to 5 micrograms/dl) are not diagnostic of adrenal insufficiency. PMID- 3038478 TI - The clinical assessment of lung water. PMID- 3038479 TI - Cytostatic and antiherpesvirus type 1 and 2 activities of 1-beta-D arabinofuranosylthymine (ara T) prodrugs. AB - A series of derivatives of the antibiotic 1-beta-D-arabinofuranosylthymine (ara T) was synthesized by esterification of the hydroxy group in the 5'-position of the arabinose moiety of the nucleoside with straight-chain and branched-chain carboxylic acids: acetyl-ara T, butyryl-ara T, valeroyl-ara T, pivaloyl-ara T and palmitoyl-ara T. These ara T prodrugs were evaluated for their effect on growth of L5178y mouse lymphoma cells and noninfected BHK-21 cells as well as for their antiviral activity in Herpes simplex virus type 1 and 2 infected BHK-21 cells. All compounds exhibited a marked antiherpes virus activity, whereas the cytostatic activity of two of them, the pivaleric ester and the palmitic ester, was extremely weak. The relative antiviral indices of the 5'-pivaloyl-ara T and 5'-palmitoyl-ara T were found to be much better than the index of ara T itself. PMID- 3038480 TI - Experience and brain development. AB - This article considers how experience can influence the developing and mature brain and proposes a new categorization scheme based upon the type of information stored and the brain mechanisms that appear to be involved in storing it. In this scheme, experience-expectant information storage refers to incorporation of environmental information that is ubiquitous in the environment and common to all species members, such as the basic elements of pattern perception. Experience expectant processes appear to have evolved as a neural preparation for incorporating specific information: in many sensory systems, synaptic connections between nerve cells are overproduced, and a subsequent selection process occurs in which aspects of sensory experience determine the pattern of connections that remains. Experience-dependent information storage refers to incorporation of environmental information that is idiosyncratic, or unique to the individual, such as learning about one's specific physical environment or vocabulary. The neural basis of experience-dependent processes appears to involve active formation of new synaptic connections in response to the events providing the information to be stored. Although these processes probably do not occur entirely independently of one another in development, the categories offer a new view more in accord with neural mechanisms than were terms like "critical" or "sensitive period." PMID- 3038481 TI - Basic concepts of CNS development. AB - Some basic principles of the development of the central nervous system (CNS) are reviewed and their implications for both normal and abnormal behavioral development are discussed. The goals of this review are: to provide a set of concepts to aid in the understanding of the variety of complex processes occurring during CNS development, to illustrate how these concepts contribute to our knowledge of the normal anatomy of the adult brain, and to delineate how modifications of normal developmental processes by traumatic injury, by environmental or experiential influences, or by genetic variations may lead to modifications in the resultant structure and function of the adult CNS. PMID- 3038482 TI - Neuronal plasticity in the mammalian brain: relevance to behavioral learning and memory. AB - Much recent activity in the neurosciences relates to the search for the brain mechanisms underlying learning and memory. In recent years a brain circuit in cerebellum and brainstem has been discovered that is responsible for the learning of a simple motor response (nictitating membrane movement). This has provided a model for neuroscientists to use in understanding the brain circuits involved in this simple form of learning and, by extension, to more complex forms ultimately, and a means of exploring the changes in neural function underlying the learning. An enduring change in neural function is represented by long-term potentiation (LTP), an alteration in synaptic efficacy seen in hippocampus and other areas. LTP can be induced experimentally and occurs as a concomitant of learning. We review data suggesting that different brain circuits may underlie different forms of learning and memory. Several current theories of learning and memory with respect to hippocampal and other brain circuit involvement are considered. We conclude with the behavioral and physiological effects of exposure to teratogens or toxins and the CNS alterations associated with dementia. PMID- 3038483 TI - Memory development and neurophysiology: accomplishments and limitations. AB - The review of Teyler and Fountain offers developmental psychologists an update on the status of neurophysiological theorizing and findings related to memory and learning. Recent findings of LTP and of localization of different types of memory in different brain areas have potential for enriching our understanding of memory development. However, we note several limitations in Teyler and Fountain's presentation in that they do not: distinguish between learning and memory, nor between storage and retrieval; address the role of knowledge-based or top-down influences in memory and learning; employ concepts that can accommodate such developmental phenomena as stages in the hierarchical reorganization of memory. We conclude that even when the age-old search for the "engram" is accomplished, these issues will remain, and that different levels of neural modeling will be required to accommodate them. It is important for developmental psychologists and neuroscientists to maintain communication for the purpose of mutual refinement of models as their knowledge bases continue to grow. PMID- 3038484 TI - [Tumors of the anterior mediastinum--surgical treatment and prognosis]. AB - This paper reports on 96 mediastinal tumors situated exclusively or mainly in the anterior mediastinum. Early surgical intervention is always indicated to establish the diagnosis and to ensure as radical a tumor resection as possible. The surgical approach depends on the tumor size and localization and is made either by sternotomy or thoracotomy. The prognosis depends on the histological classification of the tumors: out of 30 patients with lymphomas, 19 survived as compared to 10 out of 21 patients with thymoma and 15 out of 18 patients with mesenchymal tumors. Of 16 patients with endothoracic and retrosternal goiters, none died of a cause connected with the goiter and nine out of 11 patients with mesodermal tumors survived. The indication for surgery should be established extensively, since one third of the tumors is malignant and the surgical lethality (1.8%) is very low. PMID- 3038485 TI - [Primary malignant fibrous histiocytoma of the liver]. PMID- 3038486 TI - AluI-resistant chromatin of chromosome 18: classification, frequencies and implications. AB - The pericentric chromatin of chromosome 18 was found to be far more heterogeneous for restriction endonuclease AluI (5'...AG decrease CT...3') than previously thought. The extent of such heterogeneity was characterized using 50 normal Caucasians and 5 cases of trisomy 18 or Edwards' Syndrome. The AluI-resistant chromatin can arbitrarily be classified into at least five sizes by comparison with the length of the short arm (p) of chromosome 18. They are: negative (1), small (2), medium (3), large (4) and very large (5) with incidences of 11.30%, 19.13%, 29.57%, 29.57% and 10.43%, respectively. In addition the location of the chromatin can be classified into four types depending upon the position relative to the primary constriction. For example: Type I (absent); Type II (present on p arm only); Type III (present on q arm only); Type IV (present on centromere and extending into both p and q arms). The incidences of types I, II, III, and IV were 11.30%, 62.61%, 0.87%, and 25.22%, respectively. Based on limited data, AluI resistant chromatin was found to be predominantly "large" and "very large" in Edwards' Syndrome samples. In addition, no case with negative Alu-resistant chromatin was noted. Therefore, it is tempting to speculate that the amount of chromatin present on the centromere might play a role in non-disjunction in Edwards' Syndrome cases. Although the variation observed in the present study is continuous, the proposed classification has some important implications for future investigations. PMID- 3038487 TI - Saturable accumulation of retinoic acid in neural and neural crest derived cells in early embryonic development. AB - Retinoic acid (RA) binds to a cytosolic protein distinguishable from the cellular retinol (R) binding protein. Recent studies, showing an influence by R and RA on genomic expression, suggest an interaction with the cell nucleus mediated by the specific binding proteins in a manner resembling that of steroid hormones. RA can irreversibly stimulate in vitro differentiation of teratocarcinoma cells and support early embryonic development in vitamin A depleted animals. This study demonstrates a saturable, highly specific and regional accumulation of RA in the neuroepithelium and developing CNS that occurs in early but not in late fetal development in the mouse. The results suggest that a binding protein, or some other cellular mechanism for accumulation of RA is expressed in the neural cells only during restricted periods of development. High levels are recorded also in regions where cranial neural crest cells are known to migrate, and later in the visceral arches and maxillary areas, the mesenchyme of which is known to be partly derived from migrating cranial neural crest cells. The specific accumulation of RA in embryonic neural and cranial neural crest cells is in line with animal experiments and human clinical data, showing that retinoids specifically impair CNS, eye, ear, and facial development. PMID- 3038489 TI - Effect of variation in diet on lipoprotein metabolism in patients with diabetes mellitus. PMID- 3038490 TI - [The use of the molecular hybridization method for detecting the RNA of the hepatitis A virus]. PMID- 3038488 TI - Diabetes and lipoprotein receptors. PMID- 3038491 TI - [The family of human Na+,K+-ATPase genes. At least 5 genes and/or pseudogenes homologous to alpha-subunit]. PMID- 3038492 TI - [An experimental study of myocardial perfusion imaging using contrast echo aortography]. PMID- 3038493 TI - Comment: papillomaviruses not in herpes group; fluorouracil in condyloma. PMID- 3038494 TI - Influence of wheat bran and of a bulk-forming ispaghula cathartic on the bioavailability of digoxin in geriatric in-patients. AB - The present randomized study compared the influence of a gel-forming wheat bran with a nongel-forming bulk cathartic (an ispaghula formulation, Vi-Siblin S) on the steady state concentrations of digoxin in plasma in 30 geriatric in-patients treated with either combination (16 with wheat bran + digoxin, 14 with ispaghula + digoxin) for 4 weeks. After 2 but not after 4 weeks, wheat bran reduced the digoxin levels, although the levels were still within the therapeutic range. Ispaghula had no influence at any time. It is concluded that neither wheat bran nor the ispaghula formulation has any clinically relevant influence on therapeutic digoxin levels in geriatric patients. PMID- 3038495 TI - [Parvovirus B19 infections. Outbreak in a medical technology educational institute]. PMID- 3038496 TI - [Chronic obstipation]. PMID- 3038497 TI - [The tumor registry as an instrument of better management of cancer patients]. PMID- 3038498 TI - Investigations on the occurrence and impact of bovine viral diarrhea (BVD) virus infections in sheep in Turkey. PMID- 3038499 TI - [How often must maternal vaccination be repeated for the prophylaxis of rotavirus and coronavirus infections (neonatal diarrhea) in the calf?]. PMID- 3038501 TI - [Cardiac amyloidosis]. PMID- 3038502 TI - [Regulation of steroid hormone genetic activity]. PMID- 3038500 TI - Sulbactam/ampicillin. A review of its antibacterial activity, pharmacokinetic properties, and therapeutic use. AB - Sulbactam is a semisynthetic beta-lactamase inhibitor which when combined with certain beta-lactam antibacterials extends their activity against bacteria that are normally resistant to the antibiotic due to production of beta-lactamases. In combination with ampicillin it extends the antibacterial activity of ampicillin to include beta-lactamase-producing strains which are otherwise resistant, including Bacteroides fragilis, and increases the susceptibility of many sensitive strains. Sulbactam is poorly absorbed after oral administration and sulbactam/ampicillin is therefore administered parenterally, although another linked sulbactam-ampicillin compound, sultamicillin, has been developed which is well absorbed after oral administration. The basic pharmacokinetic characteristics of sulbactam after parenteral administration are similar to those of ampicillin. Multiple-dose therapy with sulbactam/ampicillin is clinically and bacteriologically effective in infections of the urinary tract, skin and soft tissue, bones and joints, respiratory tract, ears, nose and throat, as well as intra-abdominal and obstetric and gynaecological infections and septicaemia. In addition, single intramuscular doses of sulbactam/ampicillin administered with oral probenecid are therapeutically effective in gonorrhoea, including infections due to penicillinase-producing and/or ampicillin-resistant Neisseria gonorrhoeae. In the prophylaxis of infectious complications of surgery sulbactam/ampicillin is superior to placebo and appears to be similar in efficacy to several alternative antibacterial regimens. Further studies involving larger numbers of patients are needed to clarify the comparative therapeutic and prophylactic efficacy of sulbactam/ampicillin and alternative antibacterial drugs. Nonetheless, sulbactam/ampicillin improves the therapeutic and prophylactic efficacy of an antibacterial of familiar safety, and must be seen as a worthwhile advance. PMID- 3038503 TI - Demonstration of calcium-dependent proteases (calpains) and thyroglobulin proteolysis in hog thyroid cytosol. AB - Ca2+-dependent neutral proteases in hog thyroid cytosol were found to digest thyroglobulin. The protease activity was divided into two peaks by DEAE-cellulose column chromatography. Peak I was eluted at 0.2 M NaCl and required only a micromolar range of Ca2+ for its 50% activation, while peak II, which was eluted at about 0.4 M NaCl, displayed little activity until the Ca2+ concentration was increased at more than 10(-4) M. Among various inhibitors used, thiol protease inhibitors (leupeptin, E-64 and monoiodoacetic acid) were the most effective, whereas a calmodulin antagonist (trifluoperazine) and serine protease inhibitors (phenylmethyl-sulfony-fluoride and pepstatin A) were not effective, indicating that these Ca2+-dependent proteases corresponded to calpains 1 and 2. Among the substrates tested, casein was the best and thyroglobulin was also a good for calpain 2. By using immunoblotting procedure with anti-thyroglobulin antibody, it has been found that calpain 2 degrades thyroglobulin to yield 67 K and 46 K thyroglobulin and further that it also degrades 40 K thyroglobulin. PMID- 3038505 TI - Early changes in parathyroid hormone response and proteoglycan synthesis of chick embryonic femur produced by exposure to 6-aminonicotinamide in ovo. AB - The mechanism of induction of micromelia in 6-day-old chick embryo by 6 aminonicotinamide (6-AN) was investigated. Six-day-old chick embryo exposed to 6 AN did not show micromelia when tenfold excess of nicotinamide over 6-AN was co administered. The ability of nicotinamide to prevent the induction of micromelia was partially offset after 4 hr of exposure to 6-AN and completely disappeared after 6 hr. The length of time necessary for the induction of micromelia was not affected by the concentration of 6-AN. These results indicate that exposure to 6 AN for only a short period of 6 hr is sufficient to commit the limb to micromelia and that cellular components involved in the induction of micromelia alter during this period. During this period, newly synthesized proteoglycan monomers typical of cartilage decreased in average molecular size, and isolated femora did not respond to parathyroid hormone (PTH) but to dibutyryl cyclic AMP to stimulate growth of cartilage in organ culture. PMID- 3038504 TI - Response of plasma adrenal steroids to synthetic ACTH under ketoconazole in man. AB - We have investigated the effect of a single oral administration of 600 mg ketoconazole, an imidazole derivative used in clinical practice as an antimycotic agent, on the response of plasma adrenocortical steroids to 1-24 ACTH in 5 normal male subjects pretreated with dexamethasone. The 2 mg of dexamethasone was administered orally at 2300 h on the preceding night, and then a rapid ACTH test was started at 0830 h. After a 1 week interval, the ACTH test was repeated in the same manner under the same dexamethasone pretreatment, but 600 mg of ketoconazole was given orally at 0500 h. The absolute plasma concentration and the increase over the basal level of each steroid after ACTH were evaluated and compared in both conditions with and without ketoconazole administration. A single ingestion of ketoconazole caused a decrease in both indices of the responsiveness of plasma dehydroepiandrosterone and a rise in the plasma level of 17 alpha hydroxypregnenolone. The response of plasma aldosterone was clearly blunted by ketoconazole administration, whereas that of plasma corticosterone was clearly increased. Ketoconazole also blocked the response of plasma cortisol with concomitantly increased responses of plasma 11-deoxycortisol and 11 deoxycorticosterone. The increased response of plasma corticosterone seemed to be likely due to the severe inhibitory action of ketoconazole on the conversion of corticosterone to aldosterone. These results imply the inhibitory effect of ketoconazole on C17-20-lyase activity and on the conversion of corticosterone to aldosterone, and suggest an inhibitory action on 11 beta-hydroxylase activity. The effects of ketoconazole on the other enzyme activities in adrenal steroid biosynthesis were also discussed. PMID- 3038506 TI - The inhibitory effects of calmodulin antagonists on TSH-stimulated thyroid hormone release by mouse thyroid lobes. AB - Calcium ions have been shown to play a mojor regulatory role in the release of various hormones from a wide variety of endocrine organs. More recently, in vitro evidence suggests that a calcium-binding protein, calmodulin, is also involved in the release of many hormones. So we examined the effects of several types of calmodulin antagonists on TSH-stimulated thyroid hormone release in vitro. Mouse thyroid lobes (one thyro-tracheal unit/tube) were incubated in Krebs-Ringer bicarbonate buffer at 37 degrees C for 4h. Free thyroxine (fT4) released in the incubation medium, thyroidal cAMP and calmodulin content were measured by RIA. TSH (5 mU/ml) and dibutyryl cAMP (DBC) (200 micrograms/ml) caused a 2-4 fold increase in thyroidal release of fT4. The stimulatory effects of TSH on fT4 release were significantly inhibited by trifluoprazine and prenylamine lactate at the concentration of 5 X 10(-5) M. More specific calmodulin antagonists, W-7 and W-13, were also shown to inhibit TSH stimulation of fT4 release at the concentration of 5 X 10(-5) M. In contrast, TSH stimulation of fT4 release was not depressed by non-specific antagonists, W-5 or W-12, at the same concentration as 5 X 10(-5) M. Further, W-13 also markedly inhibited DBC-stimulated fT4 release. Neither TSH nor PGI2 altered the thyroidal calmodulin content, dissociating with a marked increase in the cAMP concentration. These results suggest that calmodulin plays an important role in TSH-stimulated thyroid hormone release and further that this mechanism exists, at least in part, at the site subsequent to the generation of cAMP. PMID- 3038507 TI - Affinity labeling of hormone-specific proteins. PMID- 3038508 TI - Biochemistry, molecular biology, and physiology of the glucocorticoid receptor. PMID- 3038509 TI - The prevalence of asthma among inhabitants in the vicinity of a polyurethane factory in Finland. AB - Because toluene diisocyanate (TDI) is a strong sensitizer for asthma among workers in polyurethane factories, it is mostly extracted from the factory premises. The influence of such emissions on the prevalence of asthma among the people living in the vicinity of a polyurethane factory in Kouvola, Southern Finland, was studied through a questionnaire survey sent to 6807 persons living around the factory and in a control area; of these 4182 (61%) responded. In the study area near the factory there were 68 cases of asthma out of 3153 respondents (2.2%). In the control area there were 25 cases out of 1029 respondents (2.4%). The difference is insignificant (chi 2 = 0.27). Among the middle-aged the prevalence was significantly higher in the control area (chi 2 = 6.8). There was some indication of a lower asthma prevalence in the zone nearest to the factory, possibly due to its psychologically repellent effect on asthmatics, causing them to move away. Serum samples from 62 asthma patients out of 68 contacted (91%) were received and analyzed for TDI, HDI, and MDI. A positive result for the isocyanates was observed in only one patient who had been exposed in his occupation outside the factory. It was concluded that the polyurethane factory did not have a noticeable influence upon the prevalence of asthma in its surroundings. PMID- 3038510 TI - The pathogenicity of dust from a fluorite mine. AB - The pathogenicity of mixed dust from a fluorite mine was studied by animal experiments and in vitro tests. Animal experiments showed that calcium fluorite can induce only a foreign body reaction in the lungs; the fibrous nodular lesions induced by the fluorite mine dust are due mainly to its silica component. It was demonstrated that either silica or the mixed dust of a fluorite mine can stimulate pulmonary alveolar macrophages (PAMs) to release fibrogenetic factors in vitro, but calcium fluorite cannot. It was also demonstrated that having engulfed calcium fluorite, silica, or fluorite mine mixed dust, PAMs release an elastase-active substance. The authors suggest that the emphysematous lesion seen in autopsy material of pneumoconiosis of fluorite mine workers may be caused by calcium fluorite and silica. PMID- 3038512 TI - Characterization of mineral population by index particle: implication for the Stanton hypothesis. AB - Fibers from seven crocidolite samples used by M. F. Stanton et al. (J. Natl. Cancer Inst. 67, 965-975 (1981] were measured from STEM and optical images. Their masses were computed. From these data, the population was normalized to a sample mass of 1 micrograms, and the number of fibers in various size categories of a 1 microgram sample was compared to the tumor probability for each sample. The purposes of this study were (1) to evaluate errors in and the utility of population characterization by index number (log of the number of particles longer than 8 micron with widths equal to or less than 0.25 micron) and (2) to review the Stanton hypothesis in view of this analysis. The study found that in most cases the index number is a reliable population parameter. However, index numbers between 0 and 2.5 cannot be obtained. Therefore, populations of mineral fibers will fall into two groups: those without index particles, which have indeterminant index numbers, and those with index numbers between 2.5 and 6.0. Within Stanton's populations that contain index particles, where y equals logit tumor probability and x equals index number, r equals 0.307 (insignificant). Where y equals tumor probability and x equals index number, r equals 0.503 (significant). The correlation coefficients are low enough to suggest the possibility that factors other than size and shape play a role in mineral fiber carcinogenicity. PMID- 3038511 TI - Comparative lung immunotoxicity of inhaled quartz and coal combustion fly ash. AB - Some inhaled particles that are deposited in the terminal airways and alveoli clear to the lung-associated lymph nodes, and insoluble particles have a long half-life in these tissues. Because the lung-associated lymph nodes are essential in the induction of immunity after lung immunization, the accumulation of inhaled particles in these tissues could alter immune responses that develop after lung immunization. This study evaluated the effects of inhaled insoluble fly ash particles or alpha quartz on the immune functions of lung-associated lymph nodes. F344 rats were exposed for 20 days by inhalation of fly ash from a fluidized bed combustor (FBC) or a pulverized coal combustor (PCC). Rats were similarly exposed to quartz particles (Min-U-Sil). Control rats were exposed to filtered air. Groups of ten exposed and ten control rats from each group were immunized by intratracheal instillation of 10(8) sheep red blood cells at 4, 6, 40, and 52 weeks after the start of exposures. The cellularity of the lung-associated lymph nodes and antibody-mediated immunity were evaluated at 7 days after immunization. Tissues from lung and lung-associated lymph nodes were taken for histopathology. The inhalation of FBC fly ash, PCC fly ash, and quartz particles all significantly increased the number of lymphoid cells in the lung-associated lymph nodes at each of the sacrifice times. However, FBC fly ash had no significant effect on antibody immunity, while both PCC fly ash and quartz caused suppression of antibody responses at 52 weeks after the start of exposure. Histopathology data showed that exposure to quartz and PCC fly ash caused significant cellular changes in lungs and lung-associated lymph nodes, while FBC fly ash had less effect. These data indicate that an acute exposure (20 days) to relatively insoluble particles significantly increased the cellularity of the lung associated lymph nodes for up to 52 weeks after the start of the exposure, but the pulmonary toxicity of the particles inhaled appeared to influence the effects of the exposure on antibody immune responses. PMID- 3038513 TI - beta-Glucosidase isoenzymes in Epstein-Barr virus-transformed lymphoid cell lines from normal subjects and patients with type 1 Gaucher disease. AB - Lymphoid cell lines (LCL) from 3 adult patients with non-neuropathic Gaucher disease were established by Epstein-Barr virus (EBV) transformation and were investigated from the view of enzymology. Glucosylceramide-beta-glucosidase (GlcCer-beta-glucosidase) was present in soluble and particulate fraction of LCL from normal subjects and was deficient in type 1 Gaucher LCL; the deficiency of all molecular forms, shown by electrofocusing, indicates that they are coded by the same gene. The existence of two non-specific beta-glucosidases, one soluble (minor), the other membrane-bound (major), was demonstrated in leucocytes and LCL from normals; in Gaucher LCL, these were also present in a normal range. Characteristic properties of the non-specific membrane-bound beta-glucosidase were defined: lability at acidic pH and strong inhibitory effect by detergents. These properties allowed to discriminate it from the lysosomal GlcCer-beta glucosidase and to define optimal assay conditions for determination of residual GlcCer-beta-glucosidase activity in Gaucher disease, using artificial substrate, without interference of non-specific membrane-bound beta-glucosidase. These results demonstrate that EBV-transformed LCL represent an accurate model system for enzymatic studies of Gaucher disease. PMID- 3038514 TI - The internalization of nerve growth factor by high-affinity receptors on pheochromocytoma PC12 cells. AB - The internalization and subsequent fate of the two populations of nerve growth factor (NGF) receptors on pheochromocytoma PC12 cells were explored either by identifying the relative amounts and sizes of the receptors, after incubation of cells with [125I]NGF, by cross-linking with a photoreactive heterobifunctional reagent or by following the topological distribution of the cross-linked receptors with time. The ratio of the slow, high-affinity to the fast, low affinity NGF receptor decreased over a 5-h incubation with [125I]NGF in a process which did not involve proteolytic conversion of the slow to the fast receptor. During this period the cross-linked slow receptor moved from a trypsin-labile to a trypsin-stable site suggestive of internalization. In contrast, the cross linked fast NGF receptor remained trypsin sensitive for at least 2 h of incubation, indicative of a constant cell surface localization. The internalized [125I]NGF in the cross-linked slow NGF receptor was not degraded, indicating that cross-linking, by preventing the acid pH-induced dissociation of the NGF-receptor complex in the endosomes, blocks normal sorting of [125I]NGF to the lysosomes. The cross-linked receptor was not recycled to the cell surface. If this reflects the properties of the unmodified receptor then another process, possibly receptor conversion, is required to replenish slow NGF receptors in the cell surface. PMID- 3038515 TI - Localization of the mcf.2 transforming sequence to the X chromosome. AB - A transforming sequence was identified using co-transfection of DNA from the human mammary carcinoma cell line MCF-7 and of a G418 resistance gene into NIH 3T3 cells, followed by tumor formation in athymic mice. This sequence, named mcf.2, was molecularly cloned. A transforming activity resides in a cosmid clone of 42 kb. mcf.2 did not cross-hybridize with the known oncogenes tested. In situ hybridization localized it on the X chromosome, probably at q27. This localization was confirmed by hybridization to a panel of human--rodent cell line DNAs. PMID- 3038517 TI - Two different factors bind to the alpha-domain of the polyoma virus enhancer, one of which also interacts with the SV40 and c-fos enhancers. AB - Two nuclear factors from mouse 3T6 cells bind to a 22-bp segment constituting the alpha-domain of the polyoma virus enhancer. Binding of each factor can be competed out selectively by the appropriate double-stranded oligonucleotide, indicating that this binding is not strictly cooperative. Sequence homology between the two binding sites and the similar size of the protected regions may indicate that both factors, PEA1 and PEA2, are closely related. The binding site of PEA1 is centered on a sequence showing strong homology to the SV40 enhancer, the binding site of PEA2 is located immediately adjacent to it and shows a strong homology to the c-fos enhancer. Surprisingly, both SV40 and c-fos enhancers interact with PEA1, probably due to the presence of an extra base pair relative to c-fos in the PEA2 site. Factor PEA1 is probably identical to the recently described activator protein 1 (AP1). PMID- 3038516 TI - Cell type-specificity elements of the immunoglobulin heavy chain gene enhancer. AB - A strong transcriptional enhancer was created by oligomerization of a short segment from the immunoglobulin heavy chain (IgH) enhancer. This segment was analyzed in parallel for biological activity in vivo and factor binding in vitro. In transfection experiments the oligomerized segment stimulates transcription in a cell type-specific manner similar to the entire IgH enhancer. Transfections of mutants identified two sequence motifs whose integrity is required for efficient and cell type-specific activity of this enhancer. The first is a sequence suggested previously to be bound by a factor in vivo, and the second is a highly conserved decanucleotide which also occurs in Ig variable gene promoters. The ability of these two sequence motifs to bind proteins in vitro was tested by band shift assays. Under our in vitro conditions we could not detect proteins binding to the in vivo footprint region. However, we found protein factors binding to the decanucleotide. A ubiquitous form of this factor is present in every cell line analyzed. Additional variants are detected exclusively in cells where the IgH enhancer and the segment thereof are active. Elimination of the decanucleotide motif is not only a strong down mutation in vivo but also abolishes binding of all factor variants in vitro. Thus our data suggest that the two enhancer motifs analyzed are involved in positive rather than negative control of transcription. PMID- 3038518 TI - Characterization of a cell type-specific enhancer found in the human papilloma virus type 18 genome. AB - We have previously isolated long-range acting enhancer elements from the HeLa genome by functional selection. In this report, the structural and functional characteristics of one (GA1) of the enhancers are reported. By cloning various restriction fragments and by deletion mutagenesis, the activity of GA1 was located in a 230-bp region. The nucleotide sequence of GA1 and genomic Southern blot analysis indicated that GA1 is derived from human papilloma virus (HPV) 18 DNA that had been integrated into the HeLa genome. The enhancer is located in the non-coding region of the HPV 18 genome. The HPV 18 enhancer consists of two functional domains, both of which have full enhancer activity in HeLa cells. The enhancer does not contain enhancer core sequences but contains several blocks of potential Z-DNA sequence. Like SV40 and polyoma virus enhancers, the activity of the HPV 18 enhancer was repressed by adenovirus E1a products. The HPV 18 enhancer shows a narrow cell type specificity: it is active in some cervical carcinoma cell lines, but inactive in all non-cervical cells except for one neuroblastoma cell line. These results suggest that the HPV 18 enhancer plays an important role in regulation of the viral genes. PMID- 3038519 TI - Interaction of the TGGCA-binding protein with upstream sequences is required for efficient transcription of mouse mammary tumor virus. AB - A high-affinity binding site for the TGGCA-binding protein, also known as nuclear factor I, has previously been shown to reside within the mouse mammary tumor virus (MMTV) long terminal repeat. We have introduced mutations into this binding site to test the importance of this ubiquitous nuclear protein in MMTV transcription. Mutations which abolish the binding of the TGGCA protein in vitro are shown to impair strongly glucocorticoid-induced transcription from this promoter in vivo. These data demonstrate that the TGGCA-binding protein is a multifunctional DNA-binding protein, capable of serving a transcriptional role in the case of MMTV, in addition to its known involvement in the replication of adenovirus. PMID- 3038520 TI - cDNA-derived amino acid sequence of rat mitochondrial 3-oxoacyl-CoA thiolase with no transient presequence: structural relationship with peroxisomal isozyme. AB - The sorting of homologous proteins between two separate intracellular organelles is a major unsolved problem. 3-Oxoacyl-CoA thiolase is localized in mitochondria and peroxisomes, and provides a good system for the study on the problem. Unlike most mitochondrial matrix proteins, mitochondrial 3-oxoacyl-CoA thiolase in rats is synthesized with no transient presequence and possess information for mitochondrial targeting and import in the mature protein. Two overlapping cDNA clones contained an open reading frame encoding a polypeptide of 397 amino acid residues (predicted Mr = 41,868), a 5' untranslated sequence of 164 bp, a 3' untranslated sequence of 264 bp and a poly(A) tract. The amino acid sequence of the mitochondrial thiolase is 37% identical with that of the mature portion of rat peroxisomal 3-oxoacyl-CoA thiolase precursor. These results suggest that the two thiolases have a common origin and obtained information for targeting to respective organelles during evolution. Two portions in the mitochondrial thiolase that may serve as a mitochondrial targeting signal are presented. PMID- 3038521 TI - Sequence of the bovine mitochondrial phosphate carrier protein: structural relationship to ADP/ATP translocase and the brown fat mitochondria uncoupling protein. AB - A cDNA encoding the precursor of the bovine mitochondrial phosphate carrier protein has been cloned from a bovine cDNA library using a mixture of 128 different 17-mer oligonucleotides as hybridisation probe. The protein has an N terminal extension of 49 amino acids not present in the mature protein. This extension has a net positive charge and is presumed to direct the import of the protein from the cytoplasm to the mitochondrion. Comparison of the protein sequence of the mature phosphate carrier with itself, with ADP/ATP translocase and with the uncoupling protein from brown fat mitochondria shows that all three proteins contain a 3-fold repeated sequence approximately 100 amino acids in length, and that the repeats in the three proteins are related to each other. This implies that the three proteins have related three-dimensional structures and mechanisms and that they share a common evolutionary origin. The distribution of hydrophobic residues in the phosphate carrier protein suggests that each repeated 100 amino acid element is composed of two membrane-spanning alpha helices linked by an extensive hydrophilic domain. This model is similar to that first proposed for the ADP/ATP translocase and later for the brown fat mitochondria uncoupling protein. PMID- 3038522 TI - Transformation and rescue of a flightless Drosophila tropomyosin mutant. AB - In the Drosophila flightless mutant Ifm(3)3, a transposable element inserted into the alternatively spliced fourth exon of the tropomyosin I (TmI) gene prevents proper expression of Ifm-TmI, the tropomyosin isoform found in indirect flight muscle. We have rescued the flightless phenotype of Ifm(3)3 flies using P-element mediated transformation with a segment of the Drosophila genome containing the wild-type TmI gene plus 2.5 kb of 5' flanking and 2 kb of 3' flanking DNA. The inserted TmI gene is expressed with the proper developmental and tissue specificity, although its level of expression varies among the five transformed lines examined. These conclusions are based on analyses of flight, myofibrillar morphology, and TmI RNA and protein levels. A minimum of two copies of the inserted TmI gene per cell is necessary to restore flight to most of the flies in each line. We also show that the Ifm-TmI isoform is expressed in the leg muscle of wild-type flies and is decreased in Ifm(3)3 leg muscle. Homozygous Ifm(3)3 mutants do not jump. The ability to jump can be restored with a single copy of the wild-type TmI gene per cell. PMID- 3038523 TI - The yeast ubiquitin genes: a family of natural gene fusions. AB - Ubiquitin is a 76-residue protein highly conserved among eukaryotes. Conjugation of ubiquitin to intracellular proteins mediates their selective degradation in vivo. We describe a family of four ubiquitin-coding loci in the yeast Saccharomyces cerevisiae. UB11, UB12 and UB13 encode hybrid proteins in which ubiquitin is fused to unrelated ('tail') amino acid sequences. The ubiquitin coding elements of UB11 and UB12 are interrupted at identical positions by non homologous introns. UB11 and UB12 encode identical 52-residue tails, whereas UB13 encodes a different 76-residue tail. The tail amino acid sequences are highly conserved between yeast and mammals. Each tail contains a putative metal-binding, nucleic acid-binding domain of the form Cys-X2-4-Cys-X2-15-Cys-X2-4-Cys, suggesting that these proteins may function by binding to DNA. The fourth gene, UB14, encodes a polyubiquitin precursor protein containing five ubiquitin repeats in a head-to-tail, spacerless arrangement. All four ubiquitin genes are expressed in exponentially growing cells, while in stationary-phase cells the expression of UB11 and UB12 is repressed. The UB14 gene, which is strongly inducible by starvation, high temperatures and other stresses, contains in its upstream region strong homologies to the consensus 'heat shock box' nucleotide sequence. Elsewhere we show that the essential function of the UB14 gene is to provide ubiquitin to cells under stress. PMID- 3038524 TI - Cloning and sequencing of the PIF gene involved in repair and recombination of yeast mitochondrial DNA. AB - The nuclear gene PIF of Saccharomyces cerevisiae is required for both repair of mitochondrial DNA (mtDNA) and recognition of a recombinogenic signal characterized by a 26-bp palindromic AT sequence in the ery region of mtDNA. This gene has been cloned in yeast by genetic complementation of pif mutants. Its chromosomal disruption does not destroy the genetic function of mitochondria. The nucleotide sequence of the 3.5-kb insert from a complementing plasmid reveals an open reading frame encoding a potential protein of 857 amino acids and Mr = 97,500. An ATP-binding domain is present in the central part of the gene and in the carboxy-terminal region a putative DNA-binding site is present. Its alpha helix-turn-alpha helix motif is found in DNA-binding proteins such as lambda and lactose repressors which recognize symmetric sequences. Significant amino acid homology is observed with yeast RAD3 and E. coli UvrD (helicase II) proteins which are required for excision repair of damaged DNA. PMID- 3038525 TI - DNA recognition by a new family of type I restriction enzymes: a unique relationship between two different DNA specificities. AB - The DNA sequences recognized by the allelic type I restriction enzymes EcoR124 and EcoR124/3 were determined. EcoR124 recognizes 5'-GAA(N6)RTCG-3' and EcoR124/3 recognizes 5'-GAA(N7)RTCG-3'. These are typical of sequences recognized by type I recognition enzymes in that they consist of two specific domains separated by a non-specific spacer sequence. For these two enzymes, the specific sequences are identical but the length of the non-specific spacer is different. The specific domains of EcoR124/3 are thus 3.4 A further apart than those of EcoR124 and rotated with respect to each other through a further 36 degrees. PMID- 3038526 TI - Phospholipase C activation induced by noradrenaline in rat hippocampal slices is potentiated by GABA-receptor stimulation. AB - We have studied the effect of gamma-aminobutyric acid (GABA) and other GABA receptor agonists (3-aminopropanesulphonic acid and muscimol) on the noradrenaline-induced stimulation of polyphosphoinositide metabolism in rat hippocampal slices. Formation of water-soluble inositol phosphates, and polyphosphoinositide metabolism were studied in hippocampal slices prelabelled with [3H]myoinositol. Noradrenaline induced formation of inositol mono-, bis- and trisphosphate during 10 min incubation in the presence of lithium; activation of phospholipase C by noradrenaline was also reflected by the hydrolysis of polyphosphoinositides and by the increased metabolism of phosphatidylinositol. GABA-receptor agonists were unable to activate per se phospholipase C; however, when added together with a low concentration of noradrenaline, they greatly potentiated the noradrenaline-stimulated polyphosphoinositide metabolism. We conclude that GABA-receptor agonists potentiate the effect of noradrenaline on polyphosphoinositide turnover and we discuss the role of this neurotransmitter interaction in the physiology of the hippocampus. PMID- 3038527 TI - The activation antigen BLAST-2, when shed, is an autocrine BCGF for normal and transformed B cells. AB - A shed form of the membrane-bound B cell activation marker, BLAST-2 (Epstein-Barr virus cell surface, CD 23) was immune-affinity purified from Epstein-Barr virus transformed lymphoblast conditioned medium. SDS-PAGE analysis revealed a complex of two polypeptides, mol. wts 25,000 and 12,000, here termed s-BLAST-2. We show that this complex, when purified to homogeneity, can act as a growth factor for EBV-infected B lymphoblasts and normal receptor-stimulated B cell blasts. It has no effect on resting B or T cells. These data suggest that the BLAST-2 antigen has a role in autocrine B cell growth. Additionally, this complex is a co-mitogen for PHA-stimulated murine thymocytes, a property of interleukin-1. PMID- 3038528 TI - Molecular characterization of the HLA-linked steroid 21-hydroxylase B gene from an individual with congenital adrenal hyperplasia. AB - 21-Hydroxylase deficiency which causes congenital adrenal hyperplasia is one of the most common defects of adrenal steroidogenesis. There are two 21-hydroxylase genes in man, A and B, and these have been mapped to the HLA class III region. Only the 21-hydroxylase B gene is thought to be active. To understand the molecular basis of congenital adrenal hyperplasia in a patient with the salt wasting form of the disease, we cloned and characterized his single 21 hydroxylase B gene. The nucleotide sequence of this gene and a 21-hydroxylase B gene from a normal individual have been determined. Comparison of the two sequences has revealed 11 nucleotide alterations, of which two are in the 5' flanking region, four are in introns, one is in the 3' untranslated region and four are in exons. Two of the differences in exons cause codon changes, with Ser 269 and Asn-494 in the normal 21-hydroxylase B gene being converted to Thr and Ser, respectively. These amino acid substitutions may give an insight into those residues necessary for 21-hydroxylase enzymatic activity. We have also confirmed that the 21-hydroxylase A gene is a pseudogene due to three deleterious mutations in the exons. In addition, comparison of the 21-hydroxylase B gene sequence with other published sequences indicates that this microsomal cytochrome P-450 may be polymorphic. PMID- 3038529 TI - Directed Ig class switch recombination in activated murine B cells. AB - Immunoglobulin class switch recombination occurs at frequencies of up to 10%/cell/generation in activated murine B-lymphocytes. We analysed cH gene rearrangements and switch recombinations from active and inactive IgH loci of B cells activated in various ways and immortalized by cell fusion. Although about half of the IgM+ cells show rearrangement of c mu genes, the deletion of c mu is a rare event. Half of the IgG3+ and IgG1+ cells show rearrangement of c mu genes on the inactive IgH locus and the other half of the IgG+ cells have deleted c mu from both IgH loci by switch recombination. This recombination is directed to the same switch regions on both IgH loci in 60-80% of all cases. Interleukin 4 may play a critical role in programming murine B-lymphocytes for specific switch recombination. PMID- 3038531 TI - Different subfamilies of alphoid repetitive DNA are present on the human and chimpanzee homologous chromosomes 21 and 22. AB - The alphoid repeat DNA on chimpanzee chromosome 22 was compared with alphoid repeat DNA on its human homologue, chromosome 21. Hybridization of different alphoid probes under various conditions of stringency show that the alphoid repeats of chimpanzee chromosome 22 are not closely related to those of human chromosome 21. Sequence analysis of cloned dimer and tetramer EcoRI fragments from chimpanzee chromosome 22 confirm the low overall level of homology, but reveal the presence of several nucleotide changes which are exclusive to the chromosome 21 subfamily of human alphoid DNA. Southern blot analysis of alphoid repeat DNA on the chimpanzee X chromosome suggests this subfamily has been strongly conserved during and since the separation of chimpanzee and man although the two subfamilies can be distinguished on the basis of Taq I restriction fragments. PMID- 3038530 TI - Primary structure of the gene for the murine Ia antigen-associated invariant chains (Ii). An alternatively spliced exon encodes a cysteine-rich domain highly homologous to a repetitive sequence of thyroglobulin. AB - The gene for murine Ia-associated invariant (Ii) chains (Ii31 and Ii41) was characterized by sequence analysis. The gene extends over approximately 9 kb and is organized in nine exons. Exon 1 encodes the 5' untranslated region and the cytoplasmic segment, exon 2 the membrane spanning segment and adjacent amino acids and exons 3-8 the extracytoplasmic portion of Ii31. Putative promoter sequences were found upstream of the start of the coding sequence. Between exons 6 and 7 an additional, alternatively spliced exon 6b has been identified. This exon is spliced into the mRNA coding for the Ii-related Ii41 protein. Exon 6b encodes a cysteine-rich domain of 64 amino acids. It shows a remarkably high homology to the repetitive elements in thyroglobulin, a precursor for thyroid hormone. Based on this homology, it is suggested that this domain (TgR) in Tg and in Ii41 may play a role either in hormone formation or as a carrier in the transport of molecules (thyroid hormone or processed antigen respectively) between intracellular compartments. PMID- 3038533 TI - Protein IX, a minor component of the human adenovirus capsid, is essential for the packaging of full length genomes. AB - Human adenovirus type 5 (Ad5) contains a 36-kb double-stranded DNA molecule in an icosahedral capsid. Attempts to construct Ad5 insertion mutants containing DNA of more than about 105% of the genome size resulted in viral progeny in which deletions had occurred suggesting the existence of severe constraints on the size of packageable DNA molecules. To partially circumvent these constraints we used an adenovirus vector, Ad5dlE1,3, with deletions in early regions 1 (E1) and 3 for a total net reduction in genome size of 5349 bp and an expected capacity for inserts of greater than 7 kb. To use this vector efficiently we generated a circular form of dlE1,3 DNA which could be propagated as an infectious bacterial plasmid. When this plasmid was used as a recipient for inserts of various sizes it was found that its capacity was much less than expected and that dlE1,3 virion capsids could not even package DNA as large as the wt genome. Because the E1 deletion of dlE1,3 extends into the coding sequences for protein IX, a minor capsid component known to affect the heat stability of adenovirions, the possibility that absence of this polypeptide might also affect the DNA capacity of the virion was investigated. It was found that when the coding sequences for protein IX were restored the packaging capacity of the vector was also restored to that of wt virions. Thus protein IX is an essential constituent of virion capsids dispensable only for virions containing DNA of less than genomic size. PMID- 3038532 TI - Structure of an inverted duplication formed as a first step in a gene amplification event: implications for a model of gene amplification. AB - Inverted duplications have been observed to be a common feature of gene amplification in mammalian cells and appear to be generated as a primary event in the amplification process (Ford et al., 1985; Ford and Fried, 1986). The structural features of the amplified inverted duplication, containing the polyoma virus oncogene middle T-antigen, were analysed in transformed 3B rat cells. No unusual sequences such as transposition elements were detected at the site of the inversion. The inversion was generated by a simple illegitimate recombination event in which only a single nucleotide directly at the point of the inversion cannot be accounted for from the sequence of the two parental strands. Possible structural (hairpin formation) and sequence (rich AT) features may have been involved in the illegitimate recombination event at the inversion join. In the cellular DNA near one of its joins with polyoma virus DNA an unusual sequence of 198 bp composed of 99 consecutive purine-pyrimidine pairs has been detected. A model for the generation of amplified DNA containing inverted duplications is proposed. PMID- 3038534 TI - Human papillomavirus type 16 DNA cooperates with activated ras in transforming primary cells. AB - The close association of human papillomavirus type 16 DNA with a majority of cervical carcinomas implies some role for the virus in this type of cancer. To define the transforming properties of HPV-16 DNA in vitro we have now performed transfection experiments on baby rat kidney cells using HPV-16 DNA in conjunction with an activated ras gene. We have demonstrated that a 6.6-kb DNA fragment, containing the early genes of HPV-16 under the control of Moloney murine leukaemia virus long terminal repeats (MoMuLV-LTRs), cooperates with EJ-ras in transforming these cells. Both DNAs are required and neither alone is effective. The cooperating activity appears to reside in a protein or proteins derived from the E6/E7 region of the HPV-16 genome. PMID- 3038535 TI - Base substitutions in transposable element IS1 cause DNA duplication of variable length at the target site for plasmid co-integration. AB - We demonstrate that base substitutions in the IS1 sequence affect the length of the nucleotide sequence which is duplicated during IS1-mediated co-integration. IS1K, an IS1 variant present in the Escherichia coli chromosome, has seven base substitutions in its sequence as compared with that of IS1R derived from the plasmid R100. All substitutions are located in the internal region of IS1K. We have constructed plasmids containing IS1R, IS1K and hybrids between them: one contains four base substitutions causing an amino acid substitution in the insA gene and the other has three substitutions producing an amino acid substitution in the insB gene. We have isolated co-integrate plasmids formed by each IS1 and analysed nucleotide sequences of the target sites duplicated at the co integration junctions. The results show that IS1K generates duplications of 8 or 14 bp as well as 9 bp, while IS1R exclusively generates the 9-bp duplications. Both hybrid IS1s also create 8- or 7-bp target duplications in addition to 9-bp duplications. These results indicate that the base substitutions in either insA or insB are sufficient for the occurrence of unusual target duplications, suggesting that both genes are involved in the target duplication. PMID- 3038536 TI - DNA mismatch-repair in Escherichia coli counteracting the hydrolytic deamination of 5-methyl-cytosine residues. AB - Derivatives of phage M13 were constructed and used for the in vitro preparation of heteroduplex DNA molecules containing base/base mismatches that mimick DNA lesions caused by hydrolytic deamination of 5-meC residues in Escherichia coli DNA (i.e. they carry a T/G mismatch in the special sequence context provided by the recognition site -CCA/TGG-of the Dcm-methyltransferase). Upon introduction of these heteroduplex DNAs into CaCl2-treated E. coli cells, the mismatches are efficiently repaired with high bias in favour of the DNA strand containing the mismatched guanine residue. This special DNA mismatch-repair operates on fully dam-methylated DNA and is independent of gene mutH. It thus fulfills the salient requirements of a repair pathway responsible for counteracting the spontaneous hydrolytic deamination of 5-meC in vivo. The repair efficiency is boosted by a 5 methyl group present on the cytosine residue at the next-nearest position to the 5' side of the mismatched guanine. The repair is severely impaired in host strains carrying a mutation in any of the three loci dcm, mutL and mutS. PMID- 3038537 TI - Sequence specificity of DNA topoisomerase I in the presence and absence of camptothecin. AB - Previously, we have demonstrated that in Tetrahymena DNA topoisomerase I has a strong preference in situ for a hexadecameric sequence motif AAGACTTAGAAGAAAAAATTT present in the non-transcribed spacers of r-chromatin. Here we characterize more extensively the interaction of purified topoisomerase I with specific hexadecameric sequences in cloned DNA. Treatment of topoisomerase I-DNA complexes with strong protein denaturants results in single strand breaks and covalent linkage of DNA to the 3' end of the broken strand. By mapping the position of the resulting nicks, we have analysed the sequence-specific interaction of topoisomerase I with the DNA. The experiments demonstrate that: the enzyme cleaves specifically between the sixth and seventh bases in the hexadecameric sequence; a single base substitution in the recognition sequence may reduce the cleavage extent by 95%; the sequence specific cleavage is stimulated 8-fold by divalent cations; 30% of the DNA molecules are cleaved at the hexadecameric sequence while no other cleavages can be detected in the 1.6-kb fragment investigated; the sequence specific cleavage is increased 2- to 3-fold in the presence of the antitumor drug camptothecin; at high concentrations of topoisomerase I, the cleavage pattern is altered by camptothecin; the equilibrium dissociation constant for interaction of topoisomerase I and the hexadecameric sequence can be estimated as approximately 10(-10) M. PMID- 3038538 TI - Nucleotide sequence of the gene encoding the Streptomyces albus G beta-lactamase precursor. AB - A 1400-base DNA fragment, which contains the gene encoding the extracellular active-site serine beta-lactamase of Streptomyces albus G previously cloned into Streptomyces lividans [Dehottay et al. (1986) Gene 42, 31-36], was sequenced. The gene codes for a 314-amino-acid precursor, the N-terminal region of which has the characteristics of a signal peptide. The beta-lactamase as excreted by the host strain S. lividans PD6 has a ragged N-terminus, indicating either the presence of a leader peptidase of poor specificity or the action of an aminopeptidase. The primary structure (as deduced from the nucleotide sequence) was confirmed by amino acid sequencing of a 16-residue stretch at the amino terminus of the protein, a 12-residue stretch containing the active-site serine [De Meester et al. (1987) Biochem. J. 244, 427-432] and a 23-residue stretch obtained by trypsin digestion of the protein. The beta-lactamase belongs to class A, has three half cystine residues (one of which occurs on the amino side of the active-site serine) and is inactivated by thiol reagents. Putative ribosome binding site and terminator region were identified. PMID- 3038539 TI - Dietary and hormonal regulation of L-type pyruvate kinase gene expression in rat small intestine. AB - L-type pyruvate kinase is an enzyme of the glycolytic pathway whose activity and mRNA levels fluctuate in the small intestine according to dietary status. Both the enzyme activity and mRNA concentration decline during fasting and increase upon refeeding either a glucose-rich or a fructose-rich diet. Using a single strand M 13 phage complementary to L-type pyruvate kinase mRNA as probe, we determined the level of the mRNA in the small intestine of normal, adrenalectomized, thyroidectomized, diabetic and glucagon-treated or cAMP-treated animals refed either a glucose-rich or a fructose-rich diet. The specific mRNA is present in the small intestine of normal fasted rats and increases twofold and threefold on refeeding glucose and fructose respectively. However, the hormonal control of the gene expression differs according to the dietary carbohydrate. The L-type pyruvate kinase mRNA increase, induced by glucose feeding, is hormone dependent and requires the presence of thyroid hormones and insulin. In fructose fed rats a certain level of mRNA increase occurs regardless of the hormonal status of the animals, but the full induction of the mRNA by fructose requires the presence of glucocorticoids, thyroid hormones and insulin. Thus, the hormonal regulation of L-type pyruvate kinase gene expression in the small intestine is largely similar to that described in normal rat liver but the basal mRNA level and the stimulation of the mRNA increase by fructose are higher in the small intestine. PMID- 3038540 TI - Nucleotide sequence of the ARG3 gene of the yeast Saccharomyces cerevisiae encoding ornithine carbamoyltransferase. Comparison with other carbamoyltransferases. AB - The complete nucleotide sequence of the ARG3 structural gene encoding the monomer of the trimeric ornithine carbamoyltransferase (OTCase) (EC 2.1.3.3) has been determined. It consists of 338 codons with a corresponding molecular mass of 37842 Da. Comparing OTCases from Escherichia coli, yeast, Aspergillus, rat and man emphasizes peculiarities of the yeast enzyme but also brings to light an important degree of conservation between these proteins. Comparing the various OTCases with E. coli aspartate carbamoyltransferase (ATCase) (EC 2.1.3.2) confirms the evolutionary relationship previously noted between the two types of carbamoyltransferases and points to residues probably involved in catalysis and structural folding in OTCases. PMID- 3038541 TI - Ro 18-5364, a potent new inhibitor of the gastric (H+ + K+)-ATPase. AB - The sulfoxide agent Ro 18-5364 is an extremely potent and rapid inhibitor of the gastric mucosal (H+ + K+)-ATPase with an apparent Ki of 0.1 microM at pH 6. The inhibition of both enzymatic activity and vesicular proton transport in membrane preparations is concentration- and time-dependent. Comparative studies with the two enantiomers of Ro 18-5364 indicated no enantiomeric preference. Marked differences were seen between Ro 18-5364 (sulfoxide) and Ro 18-5362 (sulfide) with regard to inhibitory activity. Even at concentrations as high as 0.1 mM Ro 18-5362 failed to affect significantly (H+ + K+)-ATPase activity and associated proton translocation. Similarly, Ro 17-5380 demonstrated an apparent Ki of 20 microM for inhibition of the (H+ + K+)-ATPase whereas its reduced derivative Ro 17-4749 was inactive. Addition of a single methyl group in the pyridine moiety of Ro 18-5364 noticeably decreased the inhibitory potency. The inhibitory action of Ro 18-5364 on (H+ + K+)-ATPase activity was markedly higher at low incubation medium pH in comparison to physiological or alkaline values. The results of incorporation studies paralleled that of enzymatic inhibition. The extent of Ro 18-5364 incorporation was dependent on time, concentration, and medium hydrogen ion concentration, with a decrease in medium pH resulting in increased binding. While ATP and GTP had little effect on the binding rates, reduced lipoic acid methyl ester, mercaptoethanol and dithiothreitol were capable of displacing the radiolabel to different extents. Autoradiography of electrophoresed Ro-18-5364 labeled gastric microsomal membranes confirmed that the radiolabel was associated with polypeptides of approximately 100 kDa. The incorporation was reversed upon subjection of the membranes to reducing conditions. PMID- 3038542 TI - Thyrotropin effect on the availability of Ni regulatory protein in FRTL-5 rat thyroid cells to ADP-ribosylation by pertussis toxin. AB - Incubation of FRTL-5 rat thyroid cell membranes with [32P]NAD and pertussis toxin results in the specific ADP-ribosylation of a protein of about 40 kDa. This protein has the same molecular mass of the alpha i subunit of the adenylate cyclase regulatory protein Ni and is distinct from proteins ADP-ribosylated by cholera toxin in the same membranes. Prior treatment of FRTL-5 cells with pertussis toxin results in the ADP-ribosylation of Ni, as indicated by the loss of the toxin substrate in the ADP-ribosylation assay performed with membranes prepared from such cells. Preincubation of FRTL-5 cells with thyrotropin causes the same loss; cholera toxin has no such effect. Pertussis toxin, as do thyrotropin and cholera toxin, increases cAMP levels in FRTL-5 cells. Forskolin together with thyrotropin, cholera toxin or pertussis toxin causes a further increase in cAMP levels. Pertussis toxin and thyrotropin are not additive in their ability to increase adenylate cyclase activity, whereas both substances are additive with cholera toxin. A role of Ni in the thyrotropin regulation of the adenylate cyclase activity in thyroid cells is proposed. PMID- 3038543 TI - The mechanism of the conservation of energy of biological oxidations. PMID- 3038544 TI - Human protein binding to DNA sequences surrounding the human T-cell lymphotropic virus type-I long terminal repeat polyadenylation site. AB - The long terminal repeats (LTRs) of RNA tumor viruses, including human T-cell lymphotropic virus type I (HTLV-I), contain the control elements for expression of viral genes. Sequence-specific LTR-DNA-binding proteins could regulate viral functions. To search for such proteins we have used an in vitro non-denaturing polyacrylamide gel assay, with restriction fragments of the HTLV-I LTR and nuclear protein extracts from several HTLV-I-infected cell lines and an uninfected T-cell line, H9. Four DNA-binding activities were observed, including non-specific DNA-binding activity and at least two activities (forms I and II) which bind specifically to a HinfI restriction fragment from nucleotides +181 to +334 relative to the transcription start site. DNA-binding activities I and II were partially resolved by ion-exchange chromatography and mapped by protection experiments to two 10-20-bp blocks surrounding the polyadenylation site at +221. Of the cell lines tested, form II was abundantly found in C10/MJ, and forms I and IV were also found in C91/PL, C81-66-45, MT2 and H9 cells. PMID- 3038545 TI - Kinetic characterization of cytochrome c oxidase from Bacillus subtilis. AB - Bacillus subtilis aa3-type cytochrome c oxidase is capable of oxidizing cytochrome c from different origins. The kinetic properties of the enzyme are influenced by ionic strength. The affinity for Saccharomyces cerevisiae cytochrome c declines with increasing ionic strength whereas the Vmax remains almost constant. An increase of Vmax is observed when the enzyme is incorporated in artificial membranes. Negatively charged phospholipids allow high turnover rates of the aa3-type oxidase. The effect of ionic strength on oxidation of horse heart cytochrome c results in significant changes of both Km and Vmax. These effects can be explained by disturbances of enzyme-substrate interactions and are not related to changes in the aggregation state of the enzyme. The respiration control index of the enzyme reconstituted in artificial membranes appeared to be dependent on phospholipid composition, protein/lipid ratios and also on the external pH. The action of the ionophores nigericin and valinomycin, at various pH values, on the enzyme activity and proton-permeability measurements of the membranes indicate that both components of the proton-motive force, the membrane potential and the pH gradient, can in principle regulate enzyme activity in the reconstituted state. PMID- 3038546 TI - The role of iron in the activation of mannonic and altronic acid hydratases, two Fe-requiring hydro-lyases. AB - D-Altronate hydratase and D-mannonate hydratase belong to a class of Fe2+ requiring enzymes, but the function of iron in these enzymes is largely unknown. Methods are described for the convenient preparation of both these hydratases from Escherichia coli and studies related to metal activation are presented. The enzymes are inactive in the absence of a bivalent metal and a reducing agent such as dithiothreitol. Fe2+ at low concentrations activates the enzymes efficiently, but inhibits them over 2 mM. Furthermore Mn2+ is also capable of activating aldonic acid hydratases and appears to be a constituent of the enzyme active center. A marked synergistic activation is observed in the presence of both ions, raising the possibility that the enzyme has two binding sites for ions. Upon activation, the two aldonic acid hydratases incorporate a single Fe atom and contain no Fe-S core, in contrast to other characterized Fe-hydratases, such as aconitase or maleic acid hydratase. The incorporated iron is losely bound (with Kd about 4.5 mM and 20 mM for mannonate and altronate hydratase, respectively) and can be readily removed with EDTA. The enzymes exhibit no requirement for sulphide ions and are insensitive to thiol reagents. A first-order inhibition is observed with iron chelators and can be removed by competition with excess metal ions. No change in the absorption spectra is observed upon oxidation-reduction or activation with metals. The activated enzymes exhibit no electron paramagnetic (EPR) spectrum under anaerobic conditions; in the presence of oxygen, an intense EPR spectrum develops in Fe2+-activated samples with signal at g = 1.98, which upon reaction of the enzyme with the substrate moves into a species with signals at g = 4.15 and g = 9.07, with EPR parameters very similar to those of oxidized rubredoxins. PMID- 3038547 TI - Highly acidic glycans from sea cucumbers. Isolation and fractionation of fucose rich sulfated polysaccharides from the body wall of Ludwigothurea grisea. AB - The body wall of the sea cucumber contains high amounts of sulfated glycans, which differ in structure from glycosaminoglycans of animal tissues and also from the fucose-rich sulfated polysaccharides isolated from marine algae and from the jelly coat of sea urchin eggs. In Ludwigothurea grisea, glycans can be separated into three fractions which differ in molecular mass and chemical composition. The fraction containing a high-molecular-mass component has a high proportion of fucose and small amounts of amino sugars, whereas another fraction contains primarily a sulfated fucan. The third fraction, which represents the major portion of the sea cucumber polysaccharides, contains besides fucose, approximately equimolar proportions of glucuronic acid and amino sugars, and has a sulfate content higher than that in the other two fractions. Both D and L isomers of fucose are found in these polysaccharides, and the sulfate is linked to the O-3 position of the fucose residues. The attachment position of the sulfate groups to the glucuronic acid units and amino sugars is still undetermined. It is possible that these compounds are involved in maintaining the integrity of the sea cucumber's body wall, in analogy with the role of other macromolecules in the vertebrate connective tissue. PMID- 3038548 TI - Effects of various metabolic conditions and of the trivalent arsenical melarsen oxide on the intracellular levels of fructose 2,6-bisphosphate and of glycolytic intermediates in Trypanosoma brucei. AB - Upon differential centrifugation of cell-free extracts of Trypanosoma brucei, 6 phosphofructo-2-kinase and fructose-2,6-bisphosphatase behaved as cytosolic enzymes. The two activities could be separated from each other by chromatography on both blue Sepharose and anion exchangers. 6-phosphofructo-2-kinase had a Km for both its substrates in the millimolar range. Its activity was dependent on the presence of inorganic phosphate and was inhibited by phosphoenolpyruvate but not by citrate or glycerol 3-phosphate. The Km of fructose-2,6-bisphosphatase was 7 microM; this enzyme was inhibited by fructose 1,6-bisphosphate (Ki = 10 microM) and, less potently, by fructose 6-phosphate, phosphoenolpyruvate and glycerol 3 phosphate. Melarsen oxide inhibited 6-phosphofructo-2-kinase (Ki less than 1 microM) and fructose-2,6-bisphosphatase (Ki = 2 microM) much more potently than pyruvate kinase (Ki greater than 100 microM). The intracellular concentrations of fructose 2,6-bisphosphate and hexose 6-phosphate were highest with glucose, intermediate with fructose and lowest with glycerol and dihydroxyacetone as glycolytic substrates. When added with glucose, salicylhydroxamic acid caused a decrease in the concentration of fructose 2,6-bisphosphate, ATP, hexose 6 phosphate and fructose 1,6-bisphosphate. These studies indicate that the concentration of fructose 2,6-bisphosphate is mainly controlled by the concentration of the substrates of 6-phosphofructo-2-kinase. The changes in the concentration of phosphoenolpyruvate were in agreement with the stimulatory effect of fructose 2,6-bisphosphate on pyruvate kinase. At micromolar concentrations, melarsen oxide blocked almost completely the formation of fructose 2,6-bisphosphate induced by glucose, without changing the intracellular concentrations of ATP and of hexose 6-phosphates. At higher concentrations (3-10 microM), this drug caused cell lysis, a proportional decrease in the glycolytic flux, as well as an increase in the phosphoenolypyruvate concentrations which was restricted to the extracellular compartment. Similar changes were induced by digitonin. It is concluded that the lytic effect of melarsen oxide on the bloodstream form of T. brucei is not the result of an inhibition of pyruvate kinase. PMID- 3038549 TI - Isolation and characterization of neurokinin A, neurokinin A(3-10) and neurokinin A(4-10) from a neutral water extract of a metastatic ileal carcinoid tumour. AB - A metastasis to the right liver lobe of an argyrophil/argentaffin midgut carcinoid tumour in a patient with the classical carcinoid syndrome was examined for the presence of tachykinins other than substance P, using a specific antiserum. The extract was initially purified using SepPak cartridges, and subsequently subjected to cation-exchange chromatography on SP Sephadex C-25 which separated the immunoreactive material into two main components (components I and II). Both were further purified by anion-exchange chromatography on DEAE Sephadex A-25, and by reverse-phase fast protein liquid chromatography. Component II was identified as neurokinin A by its immunochemical and chromatographic properties and amino acid sequence analysis. Component I consisted of two molecular forms which were identified as neurokinin A(3-10) and neurokinin A(4 10) by amino acid sequence analysis. The tumour tissue contained only small amounts of the eledoisin-like peptide that has earlier been demonstrated in mammalian tissues. Although this component behaved like the nonmammalian peptide eledoisin on reverse-phase HPLC and on reverse-phase ion-pair chromatography, eledoisin-specific antiserum E2 indicated that eledoisin-like peptide is not identical to eledoisin. Neurokinin A in carcinoid tumours has an N-terminal heterogeneity; this multiplicity constitutes a further support for the hypothesis that carcinoid tumours produce a number of tachykinins which may be present in different relative amounts in individual patients and may contribute to the individual differences in symptomatology. PMID- 3038550 TI - Identification of two isoenzymes of protein phosphatase 2C in both rabbit skeletal muscle and liver. AB - Protein phosphatase 2C was isolated from rabbit skeletal muscle by a procedure that involved chromatography on DEAE-cellulose, precipitation with ammonium sulphate, gel-filtration on Sephadex G-100, affinity chromatography on thiophosphorylated myosin-P-light-chain--Sepharose and chromatography on Mono Q. The enzyme was purified about 35,000-fold and 0.3-0.4 mg was isolated from 2500 g skeletal muscle within 5 days. The final step resolved the activity into two peaks, termed protein phosphatases 2C1 and 2C2, that possessed identical substrate specificities and enzymatic properties. About 2.5-fold more protein phosphatase 2C2 was isolated than protein phosphatase 2C1. Protein phosphatases 2C1 and 2C2 migrated as single bands on SDS/polyacrylamide gels yielding apparent molecular masses of 44 kDa and 42 kDa, respectively, and the native proteins were both monomeric at pH 7.5 as judged by their elution from Sephadex G-100 and Sephacryl S200. Peptide maps of protein phosphatases 2C1 and 2C2, obtained after separate digestions with four different proteinases, were different, indicating that they are isoenzymes. Protein phosphatases 2C1 and 2C2 were purified from rabbit liver by the same procedure, and 0.2 mg (2C1 + 2C2) was isolated from 120 g hepatic tissue. Hepatic protein phosphatases 2C1 and 2C2 were also isolated in a molar ratio of about 1:2.5, and their enzymatic properties and apparent molecular masses in the presence and absence of SDS were identical to the skeletal muscle enzymes. Protein phosphatases 2C1 from muscle and liver displayed identical peptide maps, as did protein phosphatases 2C2 from these two tissues. It is concluded that the same two isoenzymes of protein phosphatase 2C are present in skeletal muscle and liver. PMID- 3038551 TI - Oscillation of ion fluxes in mammalian erythrocytes. Mechanism of oscillation. AB - The dependence of ionophore-induced oscillations in rat erythrocytes on various concentrations of A23187, FCCP and Ca2+ was analysed using ion-selective electrodes. The oscillations were shown to be independent of the extracellular concentration of carbonylcyanide p-trifluoromethoxyphenylhydrazone and Ca2+. The dependence of oscillations on the concentration A23187 was shown to be a threshold characteristic and represented by a bell-shaped curve. In the course of oscillations the redistribution of A23187 between cells and the incubation medium was demonstrated using high-speed centrifugation. A hypothesis for oscillatory state generation in erythrocytes was suggested on the basis of pH-dependent changes of the Ca2+ ionophore A23187 content in cells. According to this hypothesis the H+ concentration within the external membrane-adjacent layer serves as a causative factor for induction of cyclic desorption of A23187 molecules from the cell membrane. PMID- 3038552 TI - Blood donor selection can prevent cytomegalovirus infection after open heart surgery. AB - Thirty-four patients without IgG antibodies against cytomegalovirus (CMV) prior to open heart surgery were studied. The patients were randomized to receive blood either from unselected donors, or from donors without detectable CMV antibodies. Fresh whole blood was mainly used. Eleven patients received CMV seronegative blood only. All had an uneventful convalescence period and remained seronegative. Ten of the 23 patients who received blood from unselected donors had typical CMV disease with the onset of symptoms in the third or fourth postoperative week, and fever lasting for two to three weeks. They all had liver enzyme release and later seroconversion against CMV. Four patients were hospitalized during this period. The incidence of symptomatic CMV infection after open heart surgery was higher than usually reported among the CMV seronegative patients. Selection of blood donors who lack antibodies against CMV may be an adequate protective measure. Avoidance of fresh blood and reduced use of blood products are presumably also of importance. PMID- 3038553 TI - The need for prevention of post-transfusion cytomegalovirus infection in patients undergoing cardiac surgery. PMID- 3038554 TI - Recent achievements and present controversies in cardiopulmonary resuscitation. PMID- 3038555 TI - Life stress and hypertension. AB - Based on psycho-physiological, clinical and epidemiological studies, essential arterial hypertension is considered to be a consequence of an inadequate 'person environment fit', objectively, subjectively or both. Besides genetic predisposition, salt intake, obesity and physical inactivity, psychological factors--among them 'hyper-reactivity' of the sympathetic nervous system, predisposing behaviour patterns and stressful life-events--should be taken into account in reaching a better understanding of the causes, prediction and prevention of hypertension. It was demonstrated that a maladaptation in various functional systems, even to minor psycho-emotional stress, is an important pathogenetic link between environment, objectively defined stressors and blood pressure regulation from the earliest phases of the disease. Implications for further research and behavioural interventions, together with other lifestyle related factors, are discussed for improving the population-based health care in cardiovascular disease in the G.D.R. PMID- 3038557 TI - Iterative reconstruction of thyroidal SPECT images. AB - First SPECT results using a multiplicative iterative reconstruction algorithm are presented. The superiority of the iterative technique over filtered backprojection is demonstrated in two thyroid SPECT studies. Obvious benefits of the new reconstruction technique are better defined outlines of the imaged organ and patient body as well as negligible artificial image amplitudes outside the patient. PMID- 3038556 TI - Release and regulation of atrial natriuretic peptide (ANP). AB - Mammalian atrial cardiocytes synthesize and secrete a hormone called atrial natriuretic peptide (ANP), which causes natriuresis, diuresis and inhibition of smooth muscle contraction, aldosterone and renin release. Volume loading, vasoconstrictor agents, immersion in water, atrial tachycardia and high salt diets have been reported to increase the release of cardiac ANP, thereby suggesting that the peptide is released in response to an increase in atrial pressure. That stretch is an important stimulus for ANP release is also suggested by clinical studies demonstrating a direct correlation between secretion rate and atrial pressure. The experiments using isolated perfused rat heart provide direct evidence that distension of the right atrium stimulates the release of ANP. Pharmacological studies in the isolated heart point to roles of cytosolic calcium, the phosphoinositide system and the cyclic AMP pathway in the regulation of ANP release. The concentration of calcium in heart muscle cells, in addition to the length of the muscle fibers, depends on many factors such as the action of humoral substances, cardiac nerve activity and heart rate, which may all contribute to the regulation of ANP secretion. PMID- 3038558 TI - Non selective transport of [11C-methyl]-L-and D-methionine into a malignant glioma. AB - Images obtained by X-ray CT, brain scintigraphy (99mTc-DTPA) and positron emission tomography (PET) with [11C-methyl]-L- and D-methionine in a case of malignant glioma are presented, showing good agreement of PET and CT findings, in particular nearly identical localization of L- and D-methionine accumulation, whereas the blood brain barrier is only slightly disturbed. In a greater number of patients the amount of accumulated stereoisomers do not differ on a significant level, indicating that a raised transport rate mediated by a carrier of low stereospecifity seems to contribute substantially to the increased uptake of [11C-methyl]-L-methionine in human brain tumors. Several cerebral functions and diseases have been studied with positron emission tomography (PET), which represents a clinical tool for visualizing metabolic activities rather than morphologic lesions (Reivich et al. 1985; Mazziotta et al. 1986). With regard to the malignancy of brain tumors DiChiro et al. (1982, 1984, 1985a, b) showed a correlation between tumor grade and its glucose metabolism measured with 18F fluorodeoxyglucose. An increased uptake of [11C-methyl]-L-methionine into tumor tissue has also been described (Hubner et al. 1980; Bergstrom et al. 1983; Kubota et al. 1984; Meyer et al. 1985; Schober et al. 1986b). Bustany et al. (1981, 1983, 1985a, b, 1986) developed a model for quantitative determination of protein synthesis, postulating that methionine incorporation into protein in brain tumors correlates with grade of malignancy. We do not believe that the uptake of [11C methyl]-L-methionine mainly reflects protein synthesis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3038559 TI - Thoracic imaging with gallium-67. AB - In the past, gallium-67 imaging has undergone several ups and downs related to its clinical importance. After a period of initial enthusiasm, its role and indications are now established. At present, there are two fields of clinical interest for 67Ga-imaging: (1) mediastinal staging in bronchogenic carcinoma and the staging of malignant lymphoma; (2) assessment of activity in interstitial lung diseases, especially sarcoidosis and inflammatory lung disorders. The advantage of 67Ga-imaging is that it is highly sensitive for the detection of neoplastic and inflammatory processes, independent of anatomical barriers. Particularly with the challenge of AIDS, 67Ga-imaging will gain increasing importance in the future. The low specificity of gallium for detecting underlying disorders precludes its use as a primary diagnostic tool. Therefore, and because of the cost and radiation load, the indications for application will have to be selected very carefully. PMID- 3038560 TI - Quality of life in patients with non-small cell lung cancer treated with chemotherapy. PMID- 3038561 TI - Analysis of Epstein-Barr virus-specific and non-specific immune functions in a patient during the development of a non-Hodgkin's lymphoma. AB - A 57-year-old woman who presented with a reactive non-malignant lymphadenopathy was observed subsequently during the development of a nodular centroblastic non Hodgkin's B-cell lymphoma. The Epstein-Barr virus (EBV)-specific antibody profile and EBV-specific and non-specific cell-mediated immune functions were determined at first presentation, and at various times during progression, in order to determine whether EBV was causally involved in the lymphoma and to assess in general the patient's cell-mediated immune function. At presentation, an immunodeficient status was suggested by an EBV-specific antibody profile indicative of an activated persistent infection; high antibody titers to viral capsid antigen (VCA) and early antigens (EA), but a low level of antibodies to EBV nuclear antigen (EBNA) confirmed by lack of leukocyte migration inhibition in response to EBNA (LMI-EBNA). The number of positive cells reactive with OKIa1 monoclonal antibody was significantly depressed, as was also the natural and interferon-activated killing (NK-IAK). After emergence of the lymphoma, NK-IAK reactivity and spontaneous lymphocyte DNA synthesis augmented in parallel with an increase in the frequency of Leu-7+ blood lymphocytes. The EBV-specific cell mediated response, reflected by the outgrowth inhibition (OI) test was abolished in parallel with a decrease in the frequency of OKT3- and OKT4-positive lymphocytes. PMID- 3038562 TI - Metabolism of 7,12-dimethylbenz[a]anthracene by mouse mammary cells in serum-free organ culture medium. AB - The murine mammary gland is a prime target organ for 7,12 dimethylbenz[a]anthracene, (DMBA)-induced carcinogenesis. We analyzed the metabolism of 3H-DMBA in a cell-free microsomal activation system derived from mouse mammary microsomes and in a whole mammary organs in culture. The in vitro microsomal activation system failed to show the more polar diol derivative of DMBA after HPLC. The metabolites obtained directly from the 3H-DMBA-treated whole mammary organs, however, revealed the presence of both the diol as well as the phenolic derivatives of DMBA. Analysis of the glands and the culture medium further showed that nearly 95% of the radioactivity added to the culture medium was associated with the adipose tissue and complete solubilization of the fat pad released substantial amounts of DMBA and its metabolites. It appears that a large portion of DMBA and its metabolites remain entrapped in the adipose tissue surrounding the parenchyma. Formic acid digestion of the gland releases the DMBA and the metabolites allowing their ethylacetate extraction, and HPLC characterization. PMID- 3038563 TI - Prazosin partly blocks clonidine-induced hypotension in patients with essential hypertension. AB - Prazosin has been reported to reduce the hypotensive and/or bradycardic effect of clonidine in various animal models. Investigations in humans have given conflicting conclusions about the effectiveness of the combination of clonidine and prazosin. In patients with essential hypertension prazosin significantly reduced the hypotensive effect of intravenous clonidine, but it failed to affect the clonidine-induced bradycardia. This finding means that the combination of prazosin and clonidine is inappropriate in antihypertensive therapy. PMID- 3038564 TI - Internalization of human T lymphocyte receptors. AB - Monoclonal antibodies specific to human T lymphocyte receptors are currently being used to define the biochemical structure of these proteins as well as of functionally distinct cell subsets. Since one of the antibodies (OKT3) recognizing the T3 (CD3) receptor mimics vital physiological processes involved in the activation of the immune system and has been successfully used as a therapeutical agent, we investigated one of the mechanisms underlying this antibody-receptor interaction. Our results show that after binding of OKT3, the complex (OKT3-T3) disappears rapidly from the cell surface. Using electron microscopy, we found that this down-regulation is due to the internalization of the complex. Parallel experiments performed on the T11 (CD2) and T4 (CD4)/AIDS retrovirus receptor indicate that the same mechanism applies for the down regulation of those molecules. These data suggest that the T3, T11 and T4 receptors have a behavior comparable to other well characterized, hormonal and viral receptors; they provide information on the metabolization pathway of surface receptors and on the possible intracellular penetration of ligands like the HTLV-III/LAV agent in human T lymphocytes. PMID- 3038565 TI - Effects of nicardipine and diltiazem on alpha-adrenoceptor responses in canine saphenous veins. AB - The effects of calcium antagonists, diltiazem and nicardipine (-6.0 to -4.0 log mol/l), on the contractile responses to noradrenaline, methoxamine and BHT-920 in isolated canine saphenous vein rings, were studied with isometric tension recordings. Concentration-effect curves to the alpha-agonists were obtained in the control state and in the presence of diltiazem or nicardipine. Propranolol ( 6.0 log mol/l) was present in the bath throughout. Diltiazem had no significant inhibitory effect on the responses mediated by all three agonists. Nicardipine ( 5.0 and -4.0 log mol/l) produced a small but significant inhibitory effect on the responses to noradrenaline and methoxamine while it had no effect on the response to BHT-920. The effects of nicardipine were greatest on the responses to methoxamine. These calcium antagonists appeared to have only small post-synaptic inhibitory effects on the contractile responses to alpha-agonists in the canine saphenous vein with nicardipine exerting a greater inhibitory influence than diltiazem. PMID- 3038566 TI - Carriers for GABA and noradrenaline uptake coexist on the same nerve terminal in rat hippocampus. AB - gamma-Aminobutyric acid (GABA; 1-300 microM) increased the basal release of [3H]noradrenaline ([3H]NA) from rat hippocampal synaptosomes. The effect of GABA at low concentrations (below 10 microM) was largely bicuculline-sensitive while the sensitivity to bicuculline decreased at higher concentrations. Muscimol mimicked GABA but only below 10 microM; bicuculline antagonized the effect of muscimol. Up to 300 microM (-)baclofen did not modify [3H]NA release. The effect of GABA was potently counteracted by SK & F 89976A, SK & F 100330A and SK & F 100561, three novel inhibitors of neuronal GABA uptake. These compounds could not entirely prevent the effect of GABA, being least effective at the lowest GABA concentrations (below 10 microM) and becoming progressively more effective when the concentrations of GABA were increased. The effect of muscimol was insensitive to SK & F 89976A. The effect of 100 microM GABA was totally prevented when bicuculline and uptake inhibitor were added together to the superfusion medium. The results suggest that the basal release of [3H]NA can be enhanced by GABA through two mechanisms: GABAA receptor activation and penetration into NA terminals by a GABA uptake process. Thus a carrier for the uptake of NA and a carrier for the uptake of GABA appear to coexist on the same nerve terminal in rat hippocampus. PMID- 3038567 TI - Effects of ring fluorination on the adrenergic properties of phenylephrine. AB - The adrenergic properties of 2-, 4- and 6-fluorophenylephrine (2-FPE, 4-FPE, 6 FPE) were compared to those of phenylephrine (PE). The order of affinities of these compounds for alpha 1-adrenoceptors as determined by displacement of [3H]prazosin and [3H]WB-4101 binding to brain membranes was the same as the order of potencies for eliciting two alpha 1-adrenergic metabolic responses in guinea pig cerebral cortical synaptoneurosomes, namely the stimulation of phosphatidylinositol turnover and the potentiation of 2-chloroadenosine-induced accumulation of cyclic AMP. In all cases the order was 6-FPE greater than PE greater than 4-FPE greater than 2-FPE. The order of affinities for alpha 2 adrenoceptors as determined by displacement of binding of [3H]clonidine to brain membrane was 6-FPE greater than PE greater than or equal to 4-FPE = 2-FPE. In contrast, the order of potencies for inhibition of forskolin-stimulated adenylate cyclase activity in human platelet membranes via an alpha 2-adrenoceptor was 6 FPE approximately equal to PE greater than 4-FPE much greater than 2-FPE. The FPEs and PE were partial agonists compared to epinephrine in human platelets. The affinities of these compounds for beta-adrenoceptors as determined by displacement of binding of [3H]dihydroalprenolol to brain membranes are 2-FPE greater than PE greater than or equal to 4-FPE much greater than 6-FPE. The FPEs and PE had positive chronotropic and inotropic effects in isolated guinea-pig atria apparently through the activation of a beta-adrenoceptor, since pindolol blocked the response while prazosin did not. 6-FPE appeared less active than the other PEs in atria. In fat cell membranes, 2-FPE was more potent than PE in stimulating adenylate cyclase via a beta-adrenoceptor, while 4-FPE and 6-FPE were inactive. Both, 2-FPE and PE were partial agonists in fat cells compared to isoproterenol. Of the three FPEs, 6-FPE represents a more potent and more selective agonist for alpha-adrenoceptors compared to beta-adrenoceptors than PE, while 4-FPE and, in particular, 2-FPE are less potent and selective as alpha agonists. PMID- 3038568 TI - Prostaglandins activate phosphoinositide metabolism in rat aorta. AB - The ability of the five prostaglandins to activate phosphoinositide (PI) metabolism and to induce contraction was studied in rat aorta. All prostaglandins (PG) tested (B2, D2, E2, F1 alpha and F2 alpha) stimulated PI hydrolysis in the range of 0.01-1,000 microM (EC50 s from 0.9 to 47 microM) and elicited contractions in the range of 0.1-100 microM (EC50 s from 0.3 to 22.1 microM). The rank order potency was PGD2 greater than F2 alpha greater than F1 alpha greater than E2 greater than B2 for PI hydrolysis and PGB2 greater than F2 alpha greater than D2 greater than E2 greater than F1 alpha for contraction. The activation of PI hydrolysis by the various prostaglandins was greater than or equal to the effect of 5-hydroxytryptamine, norepinephrine, angiotensin II and [Arg8]vasopressin. The PI hydrolytic activity of PGD2, E2 and F2 alpha correlated with their ability to induce contraction. The results with PGB2 and F1 alpha showed, however, that activation of PI hydrolysis per se is not always a sufficient or necessary condition for rat aortic contraction. PMID- 3038569 TI - Effects of chronic diazepam exposure on GABA sensitivity and on benzodiazepine potentiation of GABA-mediated responses of substantia nigra pars reticulata neurons of rats. AB - This study examined the effects of chronic diazepam treatment on GABA sensitivity of substantia nigra pars reticulata neurons and on the ability of benzodiazepines to enhance GABAergic responses of these neurons in rats. Chronic diazepam exposure failed to significantly alter the sensitivity of reticulata neurons to microiontophoretically applied GABA. However, following chronic diazepam treatment for 1 day, or 1, 3 or 7-11 weeks, reticulata neurons showed tolerance to additional systemically or iontophoretically applied benzodiazepines and displayed an increased firing rate following injection of Ro 15-1788. These changes were not apparent 24 h after cessation of chronic treatment. Thus, tolerance to the effects of benzodiazepines on reticulata neurons appeared to develop after a single day of diazepam exposure and to dissipate by 24 h after cessation of treatment. When compared to our previous studies on dorsal raphe neurons, these results demonstrate regional differences in neuronal responses to chronic diazepam exposure, which may help elucidate neural systems which are involved in tolerance to the various functional aspects of benzodiazepines. PMID- 3038571 TI - ACTH increases noradrenaline release in pithed rabbits with electrically stimulated sympathetic outflow. AB - ACTH 0.03-1 microgram/kg per min i.v. increased the noradrenaline spillover rate (the rate at which endogenous noradrenaline enters into plasma) and the plasma noradrenaline concentration in pithed rabbits with electrically stimulated sympathetic outflow. ACTH 0.1 and 1 microgram/kg per min decreased the mean arterial pressure (MAP). The effects of ACTH persisted in animals treated with propranolol. Corticosterone 10 micrograms/kg per min had no effect on the neurochemical and circulatory parameters. ACTH 0.03 and 1 microgram/kg per min increased plasma corticosterone and cortisol concentrations; the two doses of ACTH had approximately the same effect. The plasma corticosterone concentration reached after infusion of corticosterone 10 micrograms/kg per min was about twice that obtained after ACTH 0.03 or 1 microgram/kg per min. In a second series of experiments, a pressor dose of noradrenaline (1 or 2 micrograms/kg per min) was infused i.v. into pithed rabbits. ACTH 0.03 and 1 microgram/kg per min decreased blood pressure and increased heart rate in these animals. The results suggest that high doses of ACTH increase noradrenaline release by an action on postganglionic sympathetic neurons. The effect is probably not mediated through adrenal steroids. In addition, ACTH seems to decrease MAP and to increase heart rate through postsynaptic vascular and myocardial effects. PMID- 3038570 TI - Alterations in the myocardial beta-adrenoceptor system of streptozotocin-diabetic rats. AB - Previous investigations in our laboratory revealed subsensitivity of right ventricular tissue, isolated from one month STZ-diabetic rats, to the inotropic effects of isoproterenol. The present study was concerned with the characterization of this subsensitivity phenomenon. Observations of supersensitivity to methoxamine accompanied by decreased responsiveness to glucagon without a change in responsiveness to forskolin suggested a specific effect of diabetes on pathways involving receptor-mediated activation of adenylate cyclase. Radioligand binding analysis further revealed a specific decrease in the population of the high affinity state of the beta-adrenoceptor. Since the high affinity receptor state is a necessary intermediate for adenylate cyclase activation and enhanced myocardial contractility, it is proposed that the specific decrease in the high affinity population of the beta-adrenoceptor contributes to myocardial subsensitivity to isoproterenol observed in the diabetic animals. It is further proposed that the decrease in receptor population is related to increases in circulating epinephrine levels which were evident in the diabetic animals. PMID- 3038572 TI - Specific agonists for neurokinin B receptors. AB - A series of analogues of the partial sequence NKB-(4-10) (H-Asp-Phe-Phe-Val-Gly Leu-Met.NH2) was prepared in an attempt to identify selective agonists for the neurokinin B receptor type. The compounds were tested in the dog carotid artery, the rabbit pulmonary artery and the rat portal vein to evaluate their affinity for the receptors of substance P, neurokinin A and neurokinin B respectively. It has been shown that the replacement of Val7 with MePhe increased significantly the affinity of NKB-(4-10) for the neurokinin B receptor and confered marked selectivity. [MePhe7]NKB-(4-10) was practically inactive as stimulant of the receptor for NKA and was a weak agonist on the receptor for SP. Such significant changes in the pharmacological spectrum of [MePhe7]NKB-(4-10) cannot be attributed to protection from metabolism and appear to be due to changes in the peptide conformation. PMID- 3038573 TI - [D-Pen2,L-Pen5]enkephalin induced analgesia in the jimpy mouse: in vivo evidence for delta-receptor mediated analgesia. AB - Jimpy (B6CBA-A W-J/A-Ta jp) mice, which are known to be deficient in neuronal cerebroside sulfate (a putative component of the mu opioid receptor), were non responsive in the tail flick test (compared to littermate controls and a standard Swiss strain of mouse) to analgesic doses of two mu opioid receptor agonists, morphine sulfate and [D-Ala2,MePhe4,Gly-ol5]enkephalin (DAGO). However, the jimpy mice responded normally (compared to controls) to the analgesic effects of the selective delta opioid receptor agonist [D-Pen2,L-Pen5]enkephalin (DPLPE). These results suggest (1) that delta opioid receptors can mediate analgesia and (2) that cerebroside sulfate is not necessary for delta opioid receptor activation. PMID- 3038574 TI - Immunocytochemical localization of the GABAA receptor in the rat brain. PMID- 3038575 TI - A polyclonal antibody raised against clonidine: a model for the specific imidazoline receptor. PMID- 3038576 TI - Characterization of post-junctional alpha-adrenoceptors in the rat isolated perfused femoral artery. AB - A comparison was made of contractile responses to alpha-adrenoceptor agonists in the rat aorta and in the rat isolated perfused femoral artery. Dose-response curves were constructed to noradrenaline (alpha 1/alpha 2), methoxamine (alpha 1 selective) and B-HT 920 (alpha 2-selective). Methoxamine behaved as a full agonist in both tissues as compared with noradrenaline, while B-HT 920 was only a partial agonist in the aorta and produced small responses in the femoral artery preparation which were not dose-dependent. pA2 or -log KB values were calculated for prazosin and idazoxan against noradrenaline and methoxamine. Similar -log KB values for prazosin against both agonists were obtained in both tissues, while idazoxan was approximately ten times more potent in the femoral artery preparation than in the aorta. These results suggest that the aorta contains a single population of alpha 1-adrenoceptors, while the perfused femoral artery preparation contains predominantly alpha 1-adrenoceptors but also a small population of alpha 2-adrenoceptors. PMID- 3038577 TI - Modulation of [3H]flunitrazepam binding to rat cerebellar benzodiazepine receptors by phosphatidylserine. AB - The influence of phosphatidylserine on ligand binding to the benzodiazepine/GABA receptor complex was assessed in rat cerebellar synaptic membranes and in a detergent-solubilized membrane preparation. Intact synaptic membranes or membranes solubilized with the zwitterionic detergent CHAPS (3-[(3 cholamidopropyl)-dimethylammonio]propanesulfonate) were incubated with a range of concentrations of phosphatidylserine for 2 h at 4 degrees C, prior to use in radioligand binding assays. Phosphatidylserine, an endogenous membrane phospholipid, facilitated the site-specific binding of [3H]flunitrazepam to synaptic membranes and CHAPS-solubilized preparations. In addition, phosphatidylserine inhibited the facilitation of [3H]flunitrazepam binding induced by either cartazolate or gamma-aminobutyric acid (GABA). Although the maximum effect (38% facilitation of [3H]flunitrazepam binding; greater than 90% inhibition of the cartazolate action) was produced using 130 microM phosphatidylserine, a significant enhancement of [3H]flunitrazepam binding could be observed upon preincubation of synaptic membranes with concentrations of phosphatidylserine as low as 5 microM. These results suggest that endogenous phosphatidylserine may play a role in the regulation of benzodiazepine/GABA receptor function, possibly through modulation of the mechanisms which functionally link the various components of this complex receptor system. PMID- 3038578 TI - Pressure-induced cardiac hypertrophy: changes in Na+,K+-ATPase and glycoside actions in cats. AB - Effects of myocardial hypertrophy caused by pressure overload on sarcolemmal Na+,K+-ATPase and positive inotropic action of strophanthidin were examined in cats. Partial ligation of the main pulmonary artery for four weeks resulted in right ventricular hypertrophy with no significant changes in left ventricular muscle. Hypertrophy was associated with a reduction in the number of active Na+,K+-ATPase units. Affinity of the remaining enzyme for [3H]ouabain was unchanged. No apparent right or left shift in dose-response curve for the positive inotropic effect of strophanthidin was observed and toxic concentrations of strophanthidin were unchanged; however, the degree of the positive inotropic effect produced by high concentrations of strophanthidin was significantly smaller in hypertrophied muscle. Moreover, decreases in developed tension rather than tachyarrhythmias was the predominant form of toxicity observed in hypertrophied muscle. These results indicate that myocardial hypertrophy reduces the number of active Na+,K+-ATPase units per milligram protein, decreases maximal positive inotropic effect of strophanthidin, and alters the prevailing form of strophanthidin toxicity. PMID- 3038579 TI - Diprenorphine has agonist activity at opioid kappa-receptors in the myenteric plexus of the guinea-pig ileum. AB - The opioid agonist and antagonist activities of diprenorphine have been tested in four in vitro bioassay preparations. Diprenorphine is an antagonist at delta receptors in the hamster vas deferens, at mu-receptors in the rat vas deferens and at kappa-receptors in the rabbit vas deferens. In the guinea-pig ileum it is an antagonist at mu-receptors and an agonist at kappa-receptors. PMID- 3038580 TI - Acute delta 9-tetrahydrocannabinol exposure alters Ca2+ ATPase activity in neuroendocrine and gonadal tissues in mice. AB - Acute administration of delta 9-tetrahydrocannabinol (THC) (50 mg/kg) at puberty (35-40 days) significantly (P less than 0.05) reduced Ca2+ ATPase activity in hypothalamic plasma membranes but increased, although not significantly, enzyme activity in hypothalamic tissue obtained from adult mice. In contrast, testicular Ca2+ ATPase activity was increased in pubertal THC-treated males, and significantly reduced in adults. Pituitary Ca2+ ATPase activity exhibited a dose related decrease after acute THC administration at 0.5, 5 or 50 mg/kg, but there were no differential effects of age. Pituitary plasma membranes obtained from THC treated males did not respond to in vitro exposure to luteinizing hormone releasing hormone (LHRH, 10(-7) M) with the marked reduction (approximately 40%) in Ca2+ ATPase activity observed in pituitaries from oil-treated controls. In addition, effects of THC appear specific for Ca2+ ATPase activity, since Mg2+ ATPase and Na+/K+ ATPase activities were not affected. These findings indicate that acute in vivo administration of THC influences Ca2+ membrane transport, in particular Ca2+ ATPase activity. These effects occur at each level of the hypothalamic-pituitary-gonadal (HPG) axis, are related to dose and developmental age at exposure, and also appear specific for Ca2+-dependent ATPase activity. Furthermore, THC exposure modulates pituitary sensitivity to LHRH receptor mediated effects on Ca2+ ATPase activity. Therefore, effects on Ca2+ membrane transport may be involved in acute THC actions on hormonal activity at these HPG sites. PMID- 3038581 TI - [Fundamental study on ataxic mice (wriggle mouse Sagami)]. AB - Wriggle mouse Sagami (WMS), a newly discovered BALB/C mouse strain, is characterized by its locomotor instability, abnormal gait pattern and neck wriggling. Although the growth of WMS mice is delayed, compared with normal BALB/C mice, the brain size corresponds to the relatively smaller body weight. In gross or histological examinations no local atrophy appears in the cerebrum, cerebellum, brain stem or spinal cord. The c-GMP level in the WMS cerebellum is decreased, but the c-AMP level is normal. The ataxic gait is not improved significantly by the administration of thyrotropin releasing hormone (TRH). These results indicate that the mechanism inducing ataxia and abnormal gait pattern in WMS may be different from those in other genetically-determined ataxic mice, e. g., Rolling mouse Nagaya (RMN), PCD, Staggerer and Reeler. PMID- 3038582 TI - Effect of corticosteroids on estradiol and testosterone secretion by granulosa cells in culture. AB - Effect of corticosteroids on testosterone and estradiol secretion of FSH-primed pig granulosa cells were studied. Cortisol, corticosterone, deoxycorticosterone, deoxycorticosterone acetate and aldosterone enhanced testosterone synthesis, whereas cortisone and dexamethasone were found without effect. Maximal production of testosterone by granulosa cells cultured in delta-4-androstenedione supplemented medium was not further enhanced by addition of DOC. On the contrary, an inhibitory effect of corticosteroids on estradiol secretion in the presence of delta 4-androstenedione was observed. The inhibitory effect of corticosteroids on aromatisation was compared with that of the known aromatase inhibitor 4-androsten 17 beta-ol-3-one acetate. The addition of 4-androsten-17 beta-ol-3-one acetate and DOC to the culture medium supplemented with delta 4-androstenedione resulted in a marked decrease of estradiol synthesis. DOC decreased dbcAMP stimulated estradiol secretion. The results support the idea that corticosteroids may act as modulators of granulosa cell steroidogenesis and thus determine, at least partially, further follicular development or atresia. PMID- 3038583 TI - Marrow donor immunity to herpes simplex virus: association with acute graft versus-host disease. AB - The interactions between humoral and cellular immunity to herpes simplex virus (HSV) in bone marrow donors, the occurrence of active HSV infections, and the development of grades II-IV acute graft-versus-host disease (GVHD) in their HLA A,B,C,DR-identical sibling recipients were studied. The absence of IgG-class HSV antibodies in the marrow donors was associated with a low incidence of GVHD: 38 of 53 recipients (72%) of marrow from HSV-seropositive donors developed GVHD versus only two of 15 recipients (13%) with HSV-seronegative donors (p = 0.0004). The cellular immunity to HSV was studied in vitro by evaluating the degree of lymphocyte proliferative responses to that virus and was also significantly associated with GVHD: 30 of 43 recipients (70%) of marrow from donors with a positive test developed GVHD versus 10 of 25 recipients (40%) of marrow from donors with a negative test (p = 0.03). The previously reported risk for GVHD attributed to donor CMV antibodies increased the risk of GVHD due to donor HSV antibodies. Of 31 recipients of marrow from donors who were both HSV- and CMV seropositive, 27 (85%) developed GVHD versus 11 of 22 recipients (50%) of marrow from HSV-seropositive but CMV-seronegative donors (p = 0.008). PMID- 3038584 TI - Effect of cytomegalovirus infection on T-lymphocytes after lymphocyte-depleted bone marrow transplantation. AB - The effect of cytomegalovirus (CMV) infection on the repopulation of the peripheral blood with T-lymphocytes was studied in recipients of lymphocyte depleted bone marrow transplants (BMT) who had hematologic and cytogenetic evidence of engraftment. Lymphocyte depletion was performed using counterflow centrifugation and resulted in a median depletion of 98.4% (range 94.4%-99.8%) of the T cells. Between 8 and 105 days after BMT, the T-cell repopulation was characterized by a relative preponderance of T cells lacking the CD3 marker and a slow repopulation of CD3+, CD4+, and CD8+ T cells. The CD8+ T cells repopulated at a faster rate in patients with CMV infection than in those not infected with CMV. At the end of the 9- to 12-month follow-up period, patients with CMV infection had normal numbers of CD4+ and CD8+ T cells but increased numbers of HNK1+ T cells. Those without CMV infection had subnormal numbers of CD4+ T cells, normal numbers of CD8+ T cells, and numbers of HNK1+ T cells that attained the upper limit of the normal range. Most of the HNK1+ T cells in both patient groups coexpressed the CD8 marker. We conclude that the occurrence of CMV infection in recipients of lymphocyte-depleted BMT is associated with an increase in the number of T cells coexpressing CD8 and HNK1, just as in recipients of nondepleted BMT. PMID- 3038585 TI - Phagocytosis of myelin sheath fragments by dendrites. AB - In serial ultrathin sections of the frog spinal cord, profiles of dendritic appearance were identified that contained myelin fragment inclusions and received synaptic contacts. In a number of cases it could be established that the inclusions were derived from adjacent myelin sheaths. It is suggested that the phenomenon may refer to a turnover of the myelin sheath in which the detached myelin fragments are eliminated by dendrites. PMID- 3038586 TI - The effects upon the activity of hand and forearm muscles of intracortical stimulation in the vicinity of corticomotor neurones in the conscious monkey. AB - Corticomotor (CM) neurones were identified in three conscious macaque monkeys by the presence of post-spike facilitation (PSF) in spike-triggered averages of e.m.g. recorded from intrinsic hand and forearm muscles during performance of a precision grip task. Post-spike effects were compared with those produced by single-pulse intracortical microstimulation (ICMS), with strengths of 4-20 microA, delivered at the site of 47 CM cells. Most muscles facilitated by a CM cell were also facilitated by ICMS at the site of the cell. ICMS effects were stronger: at 10 microA, the amplitude of ICMS-evoked facilitation was on average 2.8 times greater than PSF, and 6.9 times greater at 20 microA. Onset latency of ICMS-evoked facilitation was consistently longer (by 1.7 and 1.3 ms at 10 and 20 microA respectively) than PSF, and it is suggested that this results from the indirect, trans-synaptic excitation of CM cells by ICMS. Post-spike suppression was rarely seen (7/421 compared to 105/421 cases of PSF). In contrast, suppression and facilitation were equally common in response to ICMS. The synaptic mechanisms underlying these effects were explored in 5 anaesthetised macaque monkeys. ICMS facilitated a greater proportion of the tested muscles than did the CM cell recorded at the stimulus site. The results suggest the juxtaposition in the motor cortex of CM neurones with different 'muscle fields'. The merits of STA and ICMS for exploring cortical organisation are discussed. PMID- 3038588 TI - Perineurial sodium-potassium-ATPase activity in streptozotocin-diabetic rats. AB - Perineurial sodium-potassium-ATPase activity was estimated in streptozotocin diabetic rats and compared with that in control animals. Total ATPase activity was found to be reduced by 34% and ouabain-sensitive ATPase activity by 53%. This finding is significant in relation to the putative role of the perineurium as a metabolically active perifascicular diffusion barrier that regulates the composition of the endoneurial fluid, as is its possible relevance to the occurrence of endoneurial edema in diabetes. PMID- 3038587 TI - Axonal GABA receptors are selectively present on normal and regenerated sensory fibers in rat peripheral nerve. AB - A sucrose gap chamber was used to study the effect of gamma-aminobutyric acid (GABA) on normal and regenerating rat peripheral nerve fibers. Sciatic nerves and dorsal roots were depolarized by GABA and the GABAA receptor subtype agonist muscimol, but not by the GABAB receptor subtype agonist baclofen. Ventral root fibers were not affected by these agents, suggesting a selective presence of axonal GABA receptors on sensory fibers of mammalian peripheral nerves. Regenerating dorsal and ventral root fibers were studied 13 to 20 days following nerve crush. The regenerated dorsal root fibers were depolarized by GABA or muscimol, but the regenerated ventral root fibers were not. These results indicate that GABA receptors are selectively present on normal mammalian sensory axons, and are reestablished on regenerated sensory axons. PMID- 3038589 TI - Recovery of tactile placing after visual cortex ablation in cat: a behavioral and metabolic study of diaschisis. AB - After bilateral visual cortex ablation, cats show a transient deficit in tactile placing and a permanent deficit in visual placing of both forelimbs. Amphetamine administration (four doses, 5 mg/kg, i.p., spaced at 4-day intervals beginning 10 days after surgery) accelerated the rate of recovery of tactile placing compared with saline controls whereas visual placing was not affected. The catecholamine antagonist, haloperidol (0.4 mg/kg, i.p.), blocked the amphetamine-enhanced recovery of tactile placing. Additionally, the visual cortex lesions produced a depression of oxidative metabolism, measured by cytochrome oxidase histochemistry, in subcortical regions remote from the injury. Animals treated with amphetamine exhibited an alleviation of this metabolic depression in the superior colliculus but not in other regions. PMID- 3038590 TI - Leishmania mexicana: uptake of sodium stibogluconate (Pentostam) and pentamidine by parasite and macrophages. PMID- 3038591 TI - Experimentally induced metastases of malignant fibrous histiocytomas xenotransplanted into nude mice from an established sarcoma cell line (RFS). AB - An experimental approach for inducing metastases from xenotransplanted malignant fibrous histiocytomas in nude mice is presented. Malignant fibrous histiocytomas were generated by inoculation of an established cell line (RFS) in the back of 16 nude mice. 7 nude mice were subjected to diminution operations carried out twice and three times, respectively, 6 out of these animals developed metastases, whereas no metastases were found in the inoperated control group (9 mice). The metastasis formation is explained by sarcoma heterogeneity, some factors which might contribute to the phenomenon of tumor dissemination are discussed. PMID- 3038592 TI - Decrease of rat-liver-T4-5'-deiodinase activity during chronical isoprenaline beta-action in vivo. AB - Pure adrenergic beta-action, brought about by long-term adrenaline + regitine application in rats, was able to increase liver T4-5'-deiodinase activity for 8 h (Nauman et al. 1984a). Long-term isoprenaline application without alpha-blocking yielded contrary results. One of the compensatory mechanism to be probably blamed could have been the very high endogenous adrenaline-levels (Porta et al. 1985). Simultaneous treatment with isoprenaline and alpha-methyltyrosinemethylester did not settle the problem since even then deiodinase activity was still drastically decreased. Two more components have been found which could very well be the reason for that "paradox beta-action" in vivo, namely very low glucose and insulin levels, both of which are known to influence deiodinase activity. In vivo -contrary to in vitro--it is seemingly not possible that catecholaminergic beta action could increase the peripheral production of T3 unless general alpha blocking took place. PMID- 3038593 TI - Further characterization of the interaction of polyoma virus and simian virus 40 with embryonal carcinoma cells. AB - The host restriction imposed on polyoma virus by embryonal carcinoma cells can be circumvented by mutations in the enhancer region of the viral genome. In addition, expression of the early viral genes can be induced by differentiating cells transfected with purified viral DNA. Although no host-range mutants of SV40 have been isolated, expression of T antigen from episomal genomes can be induced by differentiating the embryonal carcinoma cells transfected with microgram quantities of DNA. Further, transient expression of T antigen can be observed in the embryonal carcinoma cells following transfection with large amounts of viral DNA. In addition, replication of the A2 strain of polyoma in F9 cells is enhancer dependent but the SV40 enhancer can functionally substitute for the polyoma enhancer. The F9 host-range mutant TT340 contains five tandem repeats of the region surrounding the origin of replication, and it requires T antigen for replication. The parental strain (Toronto) of the mutant is able to replicate at low levels in a T antigen-dependent manner in F9 cells. This strain also has an unselected host range for PCC4 cells and the mutation in TT340 required for growth on F9 cells does not alter this inherent host range. PMID- 3038594 TI - Characterization of a mutant polyoma that expresses in F9 embryonal carcinoma cells: morphology, tumorigenicity, and restriction enzyme analysis. AB - A mutant polyoma virus (TT340), which replicates in F9 embryonal carcinoma (EC) cells and contains 2500 base pairs (bp) of additional DNA located in the early noncoding region of the genome, was analyzed to determined the DNA origin of the mutant insertion. Two fragments, representing repeated units of the 2500-bp insert, were isolated from TT340, labeled, and hybridized to the parental wild type viral DNA. A BglI 500-bp unit, of which there are approximately five copies within the 2500-bp insert, contains sequences homologous to regions on the early and late side of the viral origin of replication. A HpaII 400-bp repeated fragment shows homology to sequences on the early side with little hybridization to the late side. Removal of the 2500-bp insert results in the loss of infectivity on F9 EC cells but not on 3T6 or mouse embryo fibroblasts. Insertion of the BglI 500-bp repeat element into wild-type DNA at the BglI site allows replication of the constructed virus in F9 cells. The mutant virions were tumorigenic in newborn Syrian hamsters and the morphology of the virus was that of wild-type as assayed by electron microscopy. PMID- 3038595 TI - Prevalence of enteric adenovirus from acute gastroenteritis: a five year study. AB - Adenoviruses were detected in the stools in 459 of 3932 (11.6%) pediatric children hospitalized with acute gastroenteritis from January 1981 to December 1985. Out of the 459 adenovirus specimens 325 (8.3%) were presumptively identified as enteric adenovirus both by an adenovirus genus specific ELISA and by growth characteristics of adenovirus isolates in HEp-2 cells and in 293 cells. Enteral adenoviruses were found endemic since these viruses have been found for 5 successive years. A seasonal variation in the rates of adenovirus was also observed. Comparative data of rotavirus prevalence in the same study population are reported. PMID- 3038596 TI - A study on oral rehydration therapy of diarrheal disease in western Sicily. AB - A longitudinal study to ascertain the most common therapeutic approach to diarrheal disease by general practitioners and pediatricians was carried out in Western Sicily. Data obtained showed that of 902 home-managed cases of diarrhea observed by 58 physicians during one year, 65.3% were treated with antibiotics, 8.0% with antimotility agents and 26.7% were not treated with any pharmacological agent (rehydration or diet). Although oral rehydration therapy was widely known by physicians in Western Sicily, only a few of them were willing to use it routinely as the principal and exclusive treatment. PMID- 3038597 TI - Detection of hepatitis A virus in the stools of healthy people from endemic areas. AB - One-hundred-ninety-two stool samples were tested for the presence of human Hepatitis A antigen. Sixteen of these were also evaluated for the presence of infectious virus. All samples were obtained from young and apparently healthy people from endemic areas for Hepatitis A disease. The isolation of the infectious virus from these stools demonstrates clearly the wide diffusion of the virus in these areas, and its transmission by the oral-fecal route. PMID- 3038598 TI - Retinoic acid-induced differentiation of the human promyelocytic leukemia cell line, HL-60, and fresh human leukemia cells in primary culture: a model for differentiation inducing therapy of leukemia. PMID- 3038599 TI - 'Pseudo-lymphoid' leukaemia with unusual features: ultrastructural, immunological, cytogenetic and molecular studies. AB - An unusual case of 'pseudo-lymphoid' leukaemia is described. The leukaemic cells resembled small, mature lymphocytes but lacked B- and T-cell membrane markers as well as immunoglobulin (Ig) and T-cell receptor gene rearrangements. They showed, instead, features of early myeloid cells since they expressed 2 myeloid antigens, CDW13 and My9, and displayed peroxidase activity demonstrable by electron microscopy (EM) on unfixed cells. Cytogenetic studies showed monosomy 5, t(4;17) (p12;p11), t(2;3)(p23;q14) and an abnormal chromosome 12. Abnormalities of chromosomes 4 and 5 have been previously associated with 'pseudo-lymphoid' leukaemias. This case illustrates the value of sensitive methods for the characterization of blast cells and for the precise diagnosis of leukaemias with apparent 'lymphoid' morphology. PMID- 3038600 TI - Immunologic and virologic findings in hemophiliacs do not correlate with ecto 5'nucleotidase activity of peripheral blood lymphocytes. A difference with homosexual men. AB - It has been recently demonstrated that ecto-5'nucleotidase (5'NT) activity is significantly decreased in the peripheral blood lymphocytes (PBL) of homosexual men. This paper reports a study of PBL 5'NT activity in 38 hemophiliacs at risk for the acquired immunodeficiency syndrome (AIDS). The enzyme activity was correlated to the immunologic and virologic data. T-cell subset distribution was unbalanced and directly correlated with the cumulative amount of AHF infused. PBL 5'NT activity, however, was similar to that of healthy controls. 6 patients displayed serum antibodies to the human immunodeficiency virus (HIV) but no decrease in PBL 5'NT activity. In conclusion, these data indicate that both heavily treated and seropositive as well as untreated hemophiliacs have normal PBL 5'NT activity. This striking dissimilarity between homosexual men and hemophiliacs suggests that some immunologic alterations leading to 5'NT deficiency occur in the former only. PMID- 3038601 TI - Graft-versus-leukaemia activity associated with cytomegalovirus seropositive bone marrow donors but separated from graft-versus-host disease in allograft recipients with AML. AB - To elucidate whether a relationship existed between bone marrow donor cytomegalovirus (CMV) immune status and the probability of staying in remission after transplantation, a retrospective multicentre analysis was performed in 69 patients who received allogeneic bone marrow transplantation during relapse or second remission of AML, or second remission of ALL. None of 12 AML patients with CMV seropositive donors had posttransplant relapse, in contrast to 7 of 10 AML patients with seronegative donors. Kaplan-Meier estimates of the 2-yr probability of staying in remission for the two groups were 100% and 0%, respectively (p less than 0.0005). This effect was independent of disease stage, donor and recipient age, recipient pretransplant CMV immune status and the occurrence of posttransplant CMV infection in recipients, and was not mediated through an increased occurrence of overt graft-versus-host disease (GvHD) in recipients with CMV seropositive donors. The increased probability of staying in remission was associated with an increased probability of 3-yr disease-free survival (p less than 0.01). No similar effect was observed in patients with ALL. This study may suggest an allograft-versus-leukaemia effect in AML, associated with CMV seropositive donors, which seems separate from GvHD and independent of the occurrence of posttransplant CMV infection. PMID- 3038602 TI - Myeloid regeneration after bone-marrow transplantation monitored by serum measurements of myeloperoxidase, lysozyme and lactoferrin. AB - Bone-marrow regeneration after chemo- and radiotherapy-induced aplasia can be monitored by serum levels of myeloperoxidase (MPO), lysozyme (LYS) and lactoferrin (LF). In 10 patients with leukemia, serum measurements were performed before and after bone-marrow transplantation. Bone-marrow regeneration was suggested by increments in serum MPO and LYS 5 and 4 days prior to the increase in mononuclear cells (Mono) and 10 and 9 d before the increase in polymorphonuclear leukocytes (PMN) in the peripheral blood. LF started to rise 4.5 d before detectable circulating PMNs. 2 patients with early relapses of leukemia post transplantation are shown to display atypical patterns of serum MPO and LYS. We conclude that serum measurements of MPO, LYS and LF may be used as early and sensitive means to monitor bone-marrow activity during hematological regeneration. However, the findings also strongly support the earlier proposal that MPO alone may be used to reflect myeloid activity in the bone-marrow in general. PMID- 3038603 TI - [Molecular mechanisms of the action of 1,25-dihydroxyvitamin D3]. PMID- 3038605 TI - Detection of Ca2+-dependent cyclic GMP binding protein in frog rod outer segments. AB - For the identification of the cGMP-sensitive ion channel protein of frog rod outer segments (ROS), we analyzed cGMP binding proteins in the ROS by photoaffinity labeling with [3H]cGMP. We found three cGMP binding polypeptides (66 kDa, 92 kDa and 100 kDa) in the membrane protein fraction of ROS. cGMP binding to the 66 kDa polypeptide required the addition of 2 mM CaCl2. We propose that this polypeptide corresponds to the cGMP-activated channel protein reported by Cook et al. [(1987) Proc. Natl. Acad. Sci. USA 84, 585-589]. The 100 kDa and 92 kDa polypeptides are subunits of the cGMP phosphodiesterase. PMID- 3038604 TI - [Importance of opiate receptor ligands in regulating lymphocyte proliferative activity]. AB - Effects of morphine (10(-12)-10(-6) M), metenkephalin (10(-12)-10(-6) M), beta endorphin (10(-12)-10(-6) M), dalargin--synthetic analogue of leu-enkephalin (10( 12)-10(-7) M) and naloxone (10(-8)-10(-6) M) on lymphocytes of the human peripheral blood stimulated by polyclonal mitogens were studied. It was shown that morphine (10(-8)-10(-6) M) and dalargin (10(-12)-10(-7) M) inhibited the lymphocyte proliferative activity induced by optimal doses of phytohemagglutinin; effects of morphine and dalargin were blocked by naloxone. It was also found that beta-endorphin (10(-11)-10(9) M) inhibited the proliferative activity of lymphocytes stimulated by pokeweed mitogen; naloxone failed to block beta endorphin effect. PMID- 3038606 TI - Benzodiazepine agonists protect a histidine residue from modification by diethyl pyrocarbonate whereas propyl beta-carboline does not. AB - The pH sensitivity of benzodiazepine binding suggests that a histidine residue may be present in, or close to the benzodiazepine binding site. This was confirmed by the selective modification of histidine residues using diethyl pyrocarbonate which was found to block both benzodiazepine and beta-carboline binding. In order to assess whether this histidine residue is located in or adjacent to the benzodiazepine and beta-carboline binding sites, experiments were performed using either benzodiazepine or beta-carboline to protect against diethyl pyrocarbonate treatment. It was found that benzodiazepine agonists, but not propyl beta-carboline protect the benzodiazepine binding sites from diethyl pyrocarbonate modification. PMID- 3038607 TI - Activity of human, bovine and porcine platelet-derived growth factor in a radioreceptor assay with human placental membrane protein. AB - Radioiodinated human platelet-derived growth factor (125I-PDGF), and unlabeled human (Hu), bovine (Bo), and porcine (Po) PDGF were used to study PDGF receptors in human term placental membrane preparations. The binding of 125I-Hu-PDGF was inhibited by unlabeled preparations. Half-maximal occupancy of the binding sites occurred at 7-9 nM of Hu-, Bo- or Po-PDGF. Scatchard analysis of the binding data indicated a single class of receptors having a Kd value of about 1.1-1.2 X 10( 10) M. Thus, the sensitive radioreceptor assay does not detect any species specificity of PDGF. PMID- 3038608 TI - Production of the constant domain of murine T-cell receptor beta-chain in Escherichia coli. AB - T-cell antigen receptor is a heterodimer of disulfide-linked alpha- and beta chains. Although the essential features of T-cell receptor seem to be rather similar to those of immunoglobulin, the amount of T-cell receptor expressed on the surface of a T-cell is not large enough to be analyzed physcio-chemically. In this study, the DNA fragment encoding 120 amino acids from the 116th to the 235th of the murine beta-chain which corresponds to the presumed constant domain was inserted into an expression vector in E. coli. A large amount of this 18 kDa protein was observed to be synthesized in E. coli, and might be a good source for the three dimensional analysis of the T-cell receptor molecule. PMID- 3038609 TI - Binding of synthetic beta-human atrial natriuretic peptide to cultured rat vascular smooth muscle cells. AB - We have studied the effects of synthetic beta-human atrial natriuretic peptide (beta-hANP), an antiparallel dimer of alpha-hANP, on receptor binding and cGMP generation in cultured rat vascular smooth muscle cells and compared the effects with those of alpha-hANP. Characteristics of temperature-dependent binding and degradation of 125I-beta-hANP were similar to those of 125I-alpha-hANP. Scatchard analysis indicated a single class of binding sites for beta-hANP with a maximal binding capacity one-half that of alpha-hANP. Parallel and antiparallel dimers were equipotent in inhibiting the binding and stimulating intracellular cGMP formation, of which the maximal effect was about one-half that of alpha-hANP. Reverse-phase high performance liquid chromatography revealed that most of beta hANP added to cells was converted to a small molecular mass component corresponding to alpha-hANP after incubation. These data suggest that the less potent effect of beta-hANP in receptor binding and cGMP generation may be partly accounted for by the possible conversion of beta-hANP to alpha-hANP at the site of target cells. PMID- 3038610 TI - Evidence for two H2O2-binding sites in ferric cytochrome c oxidase. Indication to the O-cycle? AB - H2O2 addition to the oxidized cytochrome c oxidase reconstituted in liposomes brings about a red shift of the Soret band of the enzyme and an increased absorption in the visible region with two distinct peaks at approximately 570 and 605 nm. Throughout pH range 6-8.5, the spectral changes at 570 nm and in the Soret band titrate with very similar pH-independent Kd values of 2-3 microM. At the same time, Kd of the peroxide complex measured at 605 nm increases markedly with increased H+ activity reaching the value of 18 +/- 2 microM at pH 6.0. This finding may indicate the presence of two different H2O2-binding sites in the enzyme with different affinity for the ligand at acid pH. The Soret and 570 nm band effects are suggested to report H2O2 coordination to heme iron of alpha 3, whereas the maximum at 605 nm could arise from H2O2 binding to Cu alpha 3 followed by the enzyme transition into the 'pulsed' (or '420/605') conformation. Possible implication of the two H2O2-binding sites for the cytochrome oxidase redox and proton-pumping mechanisms are discussed. PMID- 3038612 TI - Pertussis toxin distinguishes between muscarinic receptor-mediated inhibition of adenylate cyclase and stimulation of phosphoinositide hydrolysis in Flow 9000 cells. AB - Pretreatment of human embryonic pituitary tumour cells (Flow 9000) with pertussis toxin significantly reduced carbachol-mediated inhibition of isoprenaline and prostaglandin E2 stimulation of cyclic AMP formation. This is in accord with an action on the inhibitory Gi-protein by pertussis toxin. In contrast, pertussis toxin-pretreatment had no effect on either muscarinic agonist or GTP[S] (a non hydrolysable GTP analogue) stimulation of [3H]inositol phosphate production in intact and permeabilized [3H]inositol-prelabelled Flow 9000 cells, respectively. These results suggest that muscarinic receptors are linked to the inhibition of adenylate cyclase and the stimulation of phosphoinositidase C via two different G proteins in Flow 9000 cells. PMID- 3038611 TI - A structural model for the alpha-subunit of transducin. Implications of its role as a molecular switch in the visual signal transduction mechanism. AB - Transducin is a GTP-binding protein which mediates the light activation signal from photolyzed rhodopsin to cGMP phosphodiesterase and is pivotal in the visual excitation process. Biochemical studies suggest that the T alpha subunit of transducin is composed of three functional domains, one for rhodopsin/T beta gamma interaction, another for guanine nucleotide binding, and a third for the activation of phosphodiesterase. The integration of the primary sequence of T alpha along with secondary structure, hydropathy and folding topology predictions, and a comparison with homologous proteins have led to the construction of a three-dimensional model of the T alpha subunit. A molecular mechanism which underlies the coupling action of T alpha is suggested on the basis of this model. PMID- 3038613 TI - Angiotensin I-converting enzyme in isolated human glomeruli. AB - Angiotensin I-converting enzyme (ACE) activity was measured with hippurylhistidylleucine as a substrate in isolated human glomeruli. The mean level was 2.2 +/- 0.47 mIU/mg glomerular protein. S9780, a newly designed competitive inhibitor of ACE, inhibited this activity by 85% at 0.3 microM. [3H]S9780 specifically bound to isolated human glomeruli. The Kd value and the number of sites were 23 nM and 83 fmol/mg, respectively. The prodrug, S9490, and Captopril were less potent than S9780 in displacing [3H]S9780 from its binding sites. Angiotensin I had no effect. Binding of [3H]S9780 was inhibited after preincubation of the glomeruli with a specific polyclonal anti-human ACE antibody. These results demonstrate that ACE is present in human adult glomeruli. PMID- 3038614 TI - Comment: CO III gene in bacteria. 'Homology between bacterial DNA and bovine mitochondrial DNA encoding cytochrome c oxidase subunit III'. PMID- 3038615 TI - Interleukin-2 binding to activated human T lymphocytes triggers generation of cyclic AMP but not of inositol phosphates. AB - Human T lymphocytes stimulated with phytohaemagglutinin undergo a single round of cell division. Further proliferation is dependent on the lymphokine interleukin-2 (IL2) [(1987) Immunology 60, 7-12]. We show here that binding of IL2 to its receptors on the lymphocyte surface triggers the generation of cyclic AMP. In contrast, generation of inositol phosphates from the breakdown of inositol lipids was not detected. We suggest that cyclic AMP may play a role in the transduction of the IL2 proliferative signal in T lymphocytes. PMID- 3038616 TI - Activation of protein kinase C inhibits prostaglandin- and potentiates adenosine receptor-stimulated accumulation of cyclic AMP in a human T-cell leukemia line. AB - Accumulation of cAMP in the human T-cell leukemia cell line Jurkat was stimulated by the adenosine analogue 5'-N-ethylcarboxamidoadenosine (NECA) and by prostaglandin E2 (PGE2). Addition of two phorbol esters, PDiBu and TPA, markedly enhanced the NECA-stimulated accumulation of cAMP whereas the PGE2-stimulated cAMP accumulation was substantially reduced. The non-tumor-promoting phorbol ester, 4 alpha-PDD, had no effect on either NECA- or PGE2-stimulated cAMP accumulation. The ability of PDiBu to inhibit the effect of PGE2 and to stimulate the effect of NECA remained in the presence a low concentration of forskolin (0.3 microM), which per se increased both NECA- and PGE2-stimulated cAMP accumulation. Our results suggest that the effect of PK-C-activating drugs on receptor-mediated cAMP accumulation is entirely dependent on which receptor is being stimulated. PMID- 3038617 TI - Calcium-activated neutral protease and its endogenous inhibitor. Activation at the cell membrane and biological function. AB - The structures of calcium-activated neutral protease (CANP) and its endogenous inhibitor elucidated recently have revealed novel features with respect to their structure-function relationship and enzyme activity regulation. The protease is regarded as a proenzyme which can be activated at the cell membrane in the presence of Ca2+ and phospholipid, and presumably regulates the functions of proteins, especially membrane-associated proteins, by limited proteolysis. Protein kinase C is hydrolysed and activated by CANP at the cell membrane to a cofactor-independent form. These results are reviewed and the possible involvement of CANP in signal transduction is discussed. PMID- 3038618 TI - A mechanism for accelerated degradation of intracellular proteins after limited damage by free radicals. AB - I propose that limited free radical attack upon proteins, occurring continuously in cells, creates new N-termini (notably aspartate and glutamate) which render the proteins more susceptible to proteolysis by the ubiquitin conjugation system. I suggest that these reactions are a significant part of the previously described 'N-end' and 'PEST' rules, which indicate amino acid termini or sequences which tend to dictate short protein half-lives. I also argue that the N-end rule may apply to sequestered intracellular sites, such as mitochondria, these also being sites of radical generation. PMID- 3038619 TI - Insulin increases membrane protein kinase C activity in rat diaphragm. AB - Calcium/phospholipid-dependent protein kinase activity (protein kinase C) was identified in rat diaphragm membrane and cytosol fractions by means of in vitro phosphorylation either of histones or of a specific 87 kDa protein substrate, combined with phosphopeptide-mapping techniques. Both insulin and tumor-promoting phorbol ester treatment of the diaphragm preparations led to increased protein kinase C activity in the membrane fractions. In contrast to the phorbol ester, however, insulin did not induce a concomitant decrease in cytosolic activity, indicating that translocation of the enzyme had not taken place. Thus, insulin appears to increase specifically membrane protein kinase C activity in rat skeletal muscle, possibly through a mechanism not identical to that induced by phorbol esters. PMID- 3038620 TI - A novel cholera toxin-sensitive G-protein (Gc) regulating receptor-mediated phosphoinositide signalling in human pituitary clonal cells. AB - Recent studies have implicated that a GTP-binding protein (G-protein) is involved in the coupling of both CCK-8 and muscarinic cholinergic receptors to phosphoinositidase C (PIC) in the human embryonic pituitary cell line, Flow 9000. Pretreatment of these cells with cholera toxin, but not pertussis toxin, inhibited the stimulation of [3H]inositol phosphate production by CCK-8 and acetylcholine. These inhibitory effects of cholera toxin could not be reproduced by treating the cells with the B-subunit of cholera toxin or cAMP-generating agents such as forskolin. These data suggest the presence of a novel Gc protein which is responsible for receptor-PIC coupling in Flow 9000 cells. PMID- 3038621 TI - Evidence for modulation of cell-to-cell electrical coupling by cAMP in mouse islets of Langerhans. AB - The effects of forskolin on electrical coupling among pancreatic beta-cells were studied. Two microelectrodes were used to measure membrane potentials simultaneously in pairs of islet beta-cells. Intracellular injection of a current pulse (delta I) elicited a membrane response delta V1 in the injected cell and also a response delta V2 in a nearby beta-cell confirming the existence of cell to-cell electrical coupling among islet beta-cells. In the presence of glucose (7 mM), application of forskolin evoked a transient depolarization of the membrane and electrical activity suggesting that the drug induced a partial inhibition of the beta-cell membrane K+ conductance. Concomitant with this depolarization of the membrane there was a marked decrease in beta-cell input resistance (delta V2/delta I) suggesting that exposure to forskolin enhanced intercellular coupling. Direct measurements of the coupling ratio delta V2/delta V1 provided further support to the idea that forskolin enhances electrical coupling among islet cells. Indeed, application of forskolin reversibly increased the coupling ratio. These results suggest that cAMP might be involved in the modulation of electrical coupling among islet beta-cells. PMID- 3038622 TI - Cyclic AMP induces a transient alkalinization in Dictyostelium. AB - In a wide range of cell types, stimulus-response coupling is accompanied by a rise in cytoplasmic pH (pHi). It is shown that stimulation of developing Dictyostelium discoideum cells with the chemoattractant cAMP also results in a rise in pHi. About 1.5 min after stimulation, pHi starts increasing from pHi approximately 7.45 to pHi approximately 7.60, as is revealed independently by two different pH null-point methods. The rise in pHi is transient, also with a persistent stimulus, and effectively inhibited by diethylstilbestrol (DES), strongly suggesting that the rise in pHi is accomplished by the DES-sensitive plasma membrane proton pump which has been demonstrated in D. discoideum. PMID- 3038623 TI - A simple method for extraction of DNA from fungi and yeasts with anhydrous hydrogen fluoride. AB - A novel method is described for the extraction of DNAs from fungi and yeasts. Anhydrous hydrogen fluoride (HF) selectively cleaves their cell walls under mild conditions (for 5 min at 0 degrees C), enabling the effective extraction of DNAs from organisms with a cell wall. A possible mechanism for this method concerning the selective cleavage of O-glycosidic linkages in cell walls has been described previously [(1977) Anal. Biochem. 82,289-309]. The extracted DNA is intact: in fact, the yeast DNA is directly applicable for restriction analysis and transformation of Escherichia coli. PMID- 3038624 TI - Late onset congenital adrenal hyperplasia: a gynecologist's perspective. PMID- 3038627 TI - [Effect of collagenase on neuromuscular transmission in the frog]. AB - Modification of synaptic function was estimated through the action of 0.1% solution of collagenase on neuromuscular junction in frog. Electrophysiological experiments revealed a decrease of the e.p.p. quantum content on single nerve stimulation with collagenase. The latter or the product of its splitting modify the action zones of mediator secretion. The demyelination action of collagenase on nerve is discussed. PMID- 3038625 TI - Human granulosa and thecal cells secrete distinct protein profiles. AB - Granulosa and thecal cells were isolated from follicles of ovaries removed from premenopausal women who underwent salpingo-oophorectomy. The electrophoretic profiles of the [35S]methionine-radiolabeled proteins secreted by the two cell types were quite distinct and showed different major proteins. Untreated granulosa cells secreted a major radiolabeled protein with a molecular weight of 220,000, identified as fibronectin by immunoprecipitation. This protein comprised 21% of the total secreted protein; however, the intensity of labeling was reduced after treatment with 0.5 mM dibutyryladenosine 3':5'-cyclic monophosphate ([Bu]2 cAMP). Fibronectin secretion by control cultures measured by a competitive enzyme linked immunoadsorbant assay ranged from 15.1 to 16.8 micrograms/mg cellular protein/24 hours and was reduced to 30% after treatment with 0.5 mM (Bu)2 cAMP. In contrast, untreated thecal cells secreted only low levels of fibronectin and a 25,000-dalton protein contributed 20% of the total secreted radiolabeled proteins. PMID- 3038626 TI - [Analysis of the effect of noradrenaline on the response of the vas deferens of the rat to transmural stimulation]. AB - In the rat isolated vas deferens, exogenous noradrenaline (NA) in concentrations of 5 X 10(-7)-5 X 10(-6) M acting for 10-15 min produced an increase whereas in concentrations above 10(-5) M it produced a decrease of the response to transmural stimulation: the increase being due to raising of the level of its secretion produced by accumulation of NA in nerve endings as well as to augmentation of postsynaptic response to NA resulting from a decrease in its reversal neuronal uptake. The decrease in the organ response to stimulation under the effect of NA at concentrations above 1 X 10(-5) M is due to desensitization of postsynaptic alpha-adrenoceptors. PMID- 3038628 TI - [Participation of cGMP in inhibitory mechanisms of the center of amphibian lymph hearts]. AB - The effects of cAMP on generation of spontaneous rhythmic activity of lymphatic pacemaker neurons, was studied in fragments of the frog isolated perfused spinal cord. The cGMP depressed both the rhythm of the burst activity and the dibutyril cAMP effect. The depressive effect of the cGMP was reversibly blocked with furosemide. The data obtained suggest existence of relationship between GABA ergic inhibition of the pacemaker activity of lymphatic center and the cGMP concentration in pacemaker neurons. PMID- 3038629 TI - [Contractile reactions of the small intestine induced by stimulation of beta adrenoreceptors]. AB - Isopropylnoradrenaline (1-2 micrograms/ml-1) induced contractile responses of the jejunum and ileum. The contractions could occur practically simultaneously with the onset of responses of arterial vessels or their latency period (reaching up to 33 sec). Blockade of M-cholinoreceptors with atropine (0.2-0.4 mg/kg) prevented both response types whereas blockade of alpha-adrenoreceptors with phentolamine (1 mg/kg) augmented them. Preliminary infusion of acetylcholine in subthreshold concentration (10(-5)-10(-6) g/ml; 1 ml/min) into the small intestine vascular bed considerably augmented the responses to isopropylnoradrenaline. The data obtained corroborate existence of presynaptic activating beta-adrenoreceptors localized on a cholinergic neuron, in the small intestine. PMID- 3038630 TI - [Role of the adrenals in inhibition of the pituitary-adrenocortical system]. PMID- 3038632 TI - Biclonal gammopathy in a patient with lymphoproliferative disease and lung cancer. PMID- 3038631 TI - [Insulin binding to erythrocytes in chronic glucocorticoid deficiency- investigation before and during replacement therapy]. AB - The effect of chronic glucocorticoid deficiency on insulin binding to erythrocyte was evaluated in 2 cases of ACTH deficiency, 2 of Sheehan's syndrome and a case of Addison's disease before and after the replacement therapy with cortisone acetate. 75 gram oral glucose tolerance test (OGTT) was also performed. Mean fasting plasma glucose (FPG) and insulin (FIRI) before (70.2 mg/dl, 3.6 microU/ml) and after (75.4 mg/dl, 7.8 microU/ml) therapy were significantly lower than those in normal control (97.0 mg/dl, 12.0 microU/ml). The ratio of FIRI/FPG before and after treatment were also significantly lower than that in control subjects. Glucose areas during OGTT before and after treatment were not different from that in control subjects. However, insulin area before treatment was significantly lower than that in control group. There were significant increases in FIRI/FPG and insulin area, but not in FPG and glucose area before and after treatment. Insulin binding to erythrocytes before treatment (11.9 +/- 1.1%, mean +/- SD) was greater than that in normal subjects (7.7 +/- 1.9%, n = 19, p less than 0.05). It was significantly decreased and normalized to 7.8 +/- 2.0% by the treatment. Analysis of binding parameters revealed the increases in receptor concentration at high affinity site (R1) and in average affinity at empty site by average affinity profile (Ke) before treatment in comparison with control subjects. There was no significant difference in binding parameters after treatment and in control group. High R1 or low R2 observed before treatment was significantly decreased or increased after treatment, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3038633 TI - Rapid procedure for preparation of macrophage plasma membrane. AB - This report describes a simple and efficient procedure for the isolation of plasma membrane from guinea pig peritoneal macrophages. The use of polycationic beads (Affi-gel 731 beads) facilitates rapid and high-clear separation of plasma membrane within 30 min. The final plasma membrane coated beads fraction has high specific activities of marker enzymes with little contamination with mitochondrial, lysosomal or cytoplasmal markers. PMID- 3038634 TI - Ferrous iron uptake by Bifidobacterium bifidum var. pennsylvanicus: the effect of metals and metabolic inhibitors. AB - Ferrous iron uptake studies in Bifidobacterium bifidum var. pennsylvanicus were carried out in a well-defined salt solution termed "modified Hanks solution" at both high iron concentrations (LAFIUS conditions) and low concentrations (HAFIUS conditions). Various divalent metals, Mn2+, Zn2+, Ni2+ and Cu2+, inhibited iron uptake under HAFIUS conditions in a non-competitive manner, and in a pseudo competitive manner under LAFIUS conditions. Cr2+ had no effect. Co2+ inhibited iron uptake competitively under HAFIUS conditions. Metabolic affectors that inhibited iron uptake both under HAFIUS and LAFIUS conditions were: tetraphenylphosphonium chloride, diethylstilbesterol, vanadate, carbonylcyanide-m chlorophenyl-hydrazone, and a mixture of valinomycin and nigericin. Substances that stimulated iron uptake were KCl, valinomycin, and nigericin. Iron uptake under LAFIUS conditions in piperazine-buffered modified Hanks solution was higher than that in the acetate-buffered solution, and acetate inhibited iron uptake in the piperazine buffer. HAFIUS showed no difference. It is concluded that iron uptake in bifidobacteria is driven by an ATPase-dependent proton-motive force and that both the pH gradient and membrane potential are involved in this process. Mn2+, Zn2+, Ni2+, and Cu2+ may be transported via LAFIUS, but not HAFIUS. HAFIUS may transport only Co2+ in addition to Fe2+. PMID- 3038635 TI - Effects of superoxide generated in vitro on glucuronidation of benzo[a]pyrene metabolites by mouse liver microsomes. AB - Glucuronidation of benzo[a]pyrene (B[a]P) metabolites, generated in situ by oxidative pathways, was studied using mouse liver uninduced microsomes. No coupling was evident between UDP-glucuronyltransferase and oxygenation of B[a]P. UDPGA protected microsomal macromolecules against binding of reactive B[a]P metabolites. Superoxide, and other reactive oxygen species decreased both the overall B[a]P metabolism and glucuronidation of some B[a]P metabolic products, and caused more extensive binding to macromolecules; UDPGA was less protective in this condition. Peroxidation of microsomes differentially affected glucuronidation of various metabolites of B[a]P, and of various model substrates, indicating that multiple glucuronyltransferases are involved in the conjugation of hydroxylated metabolites of B[a]P. PMID- 3038636 TI - Large scale use of Spherosil ion exchangers in plasma fractionation. AB - Spherosil microbeads are spherical and made of porous silica. Their surface is coated with hydrophilic and/or hydrophobic polymers. They are specially designed for the separation of proteins on an industrial scale either by ion exchange or by bioaffinity chromatography. Since 1980, large columns have been used in Institut Merieux for the purification of placental albumin and several vaccines. Here we describe a chromatographic process for the purification of human albumin and immunoglobulins (IgG) from 25 1 of plasma per cycle. First the plasma was freed of the coagulation factors and then clarified at pH 5.25. The corresponding supernatant was filtered and processed in sterile conditions. Albumin was then purified by ion exchange on 3 successive columns, respectively containing: 6.25 kg of DEAE SPHEROSIL W-1000; 3.5 kg of QMA SPHEROSIL PH-1000; 8 kg of COOH SPHEROSIL W-1000. The concentration and pH of the buffers were selected to reduce, as much as possible, the total quantity of ion exchangers required per cycle. IgG were then purified from the filtrate of the first of the previous columns. 1 column of 6.25 kg of DEAE SPHEROSIL W-1000 was used at pH 6.8. The selected chromatographic parameters allowed us to demonstrate a total elimination of HBs Ag and HB Virus when voluntarily added to an initial sample (RIA determination and HBV-DNA analysis by molecular hybridization). A second column of a large pore anion exchanger: DEAE SPHEROSIL LP-3000 was added at the end of the IgG purification, as a final security to avoid any risk of transmitting the Hepatitis B virus. The yield and quality of the final products will be presented. PMID- 3038637 TI - On the prevention of viral transmitted infections in plasma products. AB - A short review is given on screening tests and regulatory requirements of viral inactivation for plasma products in the Federal Republic of Germany. Guidelines for blood group determination and blood transfusion and guidelines for plasmapheresis contain the particular directions for blood and plasma donations. The licensing of drugs is regulated by the Drugs Act of 1976. To prevent virus infections the following screening tests are advised: HBs antigen, ALT, and LAV/HTLV-III antibody determination. PMID- 3038638 TI - Virus safety of pasteurized factor VIII and factor IX concentrates: study in virgin patients. AB - In a long term surveillance study hemophilia A and B patients were treated with a Factor VIII HS and Factor IX HS concentrate respectively, both pasteurized by heating in solution: 10 hours at 60 degrees C. None of 31 virgin hemophiliacs treated with Factor VIII HS Behringwerke developed hepatitis B during a follow up between 6 to 60 months. One patient who received 379.280 IU Factor VIII by 977 applications showed a seroconversion after 961 days of treatment. Passive/active immunisation is suggested. 4 patients had moderate elevations of transaminases (less than 120 U/l) without clinical signs of liver disease. 2 patients suffered a non-icteric NA NB-hepatitis two months after synovectomy in the same hospital. 6 virgin hemophilia B patients who had been treated with Factor IX concentrate HS Behringwerke remained serologically negative and did not develop any symptoms indicative of a hepatic disease during a follow up between 11-29 months. The HTLV III safety of Factor VIII HS and Factor IX HS Behringwerke is presently under investigation by determination of the corresponding antibody. PMID- 3038639 TI - Clinical studies with treated clotting factor concentrates. AB - We carried out safety studies in 45 previously untreated patients with congenital coagulatory defects, who needed to be treated with clotting factor concentrates. Non-A, non-B hepatitis developed in patients who received dry heated F VIII preparations with or without chloroform, but not in those who were infused with hot-steam treated F VIII or chromatography treated F IX. Hepatitis B developed in 3 unvaccinated patients who received the same lot of hot steam treated F VIII. None of the 45 patients we have investigated developed HTLV-III/LAV antibody. Thus, dry heating of concentrates does not prevent hepatitis transmission. Hot steam and hydrophobic interaction chromatography seem to be more effective in preventing hepatitis transmission, but not completely safe. All the above procedures except hot steam seem to protect from hepatitis B. They all seem to prevent HTLV-III/LAV transmission. PMID- 3038640 TI - Further examination of ontogenetic limitations on conditioned taste aversion. AB - This study was to resolve a discrepancy in the literature as to the capability of infant rats in acquiring conditioned taste aversion. Previous studies had indicated that during the 1st postnatal week, an aversion to saccharin could be conditioned when paired with lithium chloride (LiCl). Analogous conditioning with sucrose did not seem to occur until the end of the 2nd postnatal week, however, even though sucrose is discriminated from water and preferred before then. We observed that 5- and 9-day old pups express conditioned taste aversion to both saccharin and sucrose flavors that previously were paired with illness induced by LiCl. This learning occurred only when several hours separated cannulation and conditioning. A number of other factors that seemed likely to determine this early learning were found to have no effect. Thus it appears that rats can learn taste aversions very early in life, but only under certain circumstances. The results are discussed with reference to Vogt and Rudy's (1984) conclusions on the ontogeny of taste guided behaviors in the rat. PMID- 3038641 TI - In vivo insulin effect on ATPase activities in erythrocyte membrane from insulin dependent diabetics. AB - Na+-K+-dependent ouabain-sensitive ATPase and Mg2+-ATPase have been assayed in the erythrocyte membranes of control subjects and in uncontrolled type I (insulin dependent) diabetics. A decrease in Na+-K+-ATPase activity was observed in the patients that was significantly correlated with glycemia. The Mg2+-ATPase was increased moderately, and no correlation with glycemia was found. To study the in vivo effect of insulin, ATPase activities were measured in uncontrolled diabetics before and after a 24-h continuous insulin perfusion administered by means of an artificial pancreas. ATPase activities were corrected after normalization of glycemia. It therefore seems that glycemia and/or insulinemia are involved in the regulation of erythrocyte Na+-K+ ouabain-sensitive ATPase and to a lesser extent in that of Mg2+-dependent ATPase. PMID- 3038642 TI - Altered action of glucagon on human liver in type 2 (non-insulin-dependent) diabetes mellitus. AB - Glucagon may play a role in the metabolic derangements of overt Type 2 (non insulin-dependent) diabetes mellitus. We therefore have evaluated the early steps in glucagon action by investigating the hormone-sensitive adenylyl cyclase system in liver membranes from seven Type 2 diabetic patients with fasting hyperglycaemia and two-fold elevations in plasma glucagon. The comparison was made with seven control subjects matched for age, sex and body weight. Glucagon receptor binding was almost identical in the two groups. There were, however, marked alterations in the adenylyl cyclase activity in membranes from the diabetic patients. This activity was reduced by 35-50% when compared to control activity. Basal cyclase activity, as well as the activity after stimulation with glucagon or with agents (i.e., sodium fluoride and forskolin) that act beyond the glucagon receptor, was significantly decreased (p less than 0.05, p less than 0.001 respectively). In conclusion, uncontrolled Type 2 diabetes in associated with an over-all loss of responsiveness of the hormone-sensitive adenylyl cyclase in human liver, which apparently results from post-receptor alterations. This change may provide a mechanism for reducing the effect of hyperglycagonaemia in Type 2 diabetes mellitus. PMID- 3038643 TI - cDNA cloning, sequencing and chromosome mapping of a non-erythroid spectrin, human alpha-fodrin. AB - Several overlapping cDNA clones encompassing 2760 nucleotides of the alpha subunit of a human non-erythroid spectrin (termed fodrin) were isolated from a human lung fibroblast cDNA library. DNA and RNA blot analyses indicated that a single copy alpha-fodrin gene encodes a 9-kb transcript. The cDNA clones were sequenced, and all were found to contain long open reading frames. The overlapping regions were identical except for a 60-nucleotide inframe insertion at position 1133 in the composite sequence. This result suggests that at least two distinct transcripts exist in fibroblast cells. The chromosomal location of human alpha-fodrin was assigned to 1p34-1p36.1 by hybridization to somatic cell hybrids, and it is thus distinct from that of human alpha-spectrin which has been mapped to 1q22-1q25. Alignment of the composite 919 amino acids of the predicted protein sequence of human alpha-fodrin with that of human alpha-spectrin indicated that alpha-fodrin has a similar 106-amino-acid repeating structure, which is homologous with alpha-spectrin repeats 7-15. Repeats 10 and 11 are anomalous in sequence and structure from other repeats. A comparison of nucleic acid and amino acid homologies between alpha-spectrin and the alpha-fodrin of several vertebrates indicated that human non-erythroid alpha-fodrin and the common alpha-subunit of erythroid and non-erythroid cells of non-mammalian vertebrates are closely related (90%-96% amino acid homology), whereas alpha fodrin is only distantly related to the erythroid-specific alpha-spectrin subunit of mammals (55%-59% amino acid homology). These data suggest that mammalian erythroid alpha-spectrin evolved by duplication and rapid divergence from an ancestral alpha-fodrin-like gene. PMID- 3038644 TI - [Benzodiazepines: mechanism of action, therapeutic use, adverse effects (2)]. PMID- 3038645 TI - Phospholipid activation of the insulin receptor kinase: regulation by phosphatidylinositol. AB - A soybean phospholipid mixture produced a concentration-dependent enhancement of beta subunit autophosphorylation of the detergent-soluble, purified human placental insulin receptor. Although phosphatidylcholine, phosphatidylethanolamine, or phosphatidylserine also increased insulin receptor autophosphorylation, only phosphatidylinositol (PtdIns) stimulated to a similar extent as the phospholipid mixture. The effect of PtdIns was biphasic, stimulating at low concentrations (75 microM), but having no stimulatory effect at high concentrations (1.0 mM). Phospholipids also stimulated the exogenous protein kinase activity of the insulin receptor toward histone H2B. Phosphorylation of PtdIns occurred with these purified insulin receptor preparations, but this activity was insulin-independent, and the turnover number for PtdIns phosphorylation in the presence of soybean phospholipid was 1/220th as small as the turnover number for the autophosphorylating activity. These results suggest that although PtdIns can modulate the activity of the insulin receptor kinase, PtdIns phosphorylation itself is not directly involved in this regulation. PMID- 3038646 TI - Proliferation of a human epidermal tumor cell line stimulated by urokinase. AB - Several tumor cells secrete significantly increased amounts of the plasminogen activator urokinase, a trypsinlike serine protease, whose biological function in tumor biology is unclear. In this study we report that cells of the human epidermal tumor cell line CCL 20.2 express about 80,000 high-affinity urokinase receptors per cell that bind active as well as diisopropylfluorophosphate-treated high-molecular-weight (HMW) urokinase. Low-molecular-weight (LMW) urokinase is not bound to the receptor. Occupation of these receptors by active HMW urokinase stimulates cell proliferation independently in the presence of plasminogen in the culture medium. LMW urokinase has again no effect on cell proliferation. Calculated on a molar basis, this effect is about 28% of that of epidermal growth factor. Active HMW urokinase might therefore provide an autocrine receptor mediated growth-promoting mechanism for tumor cells similar to those described for other growth factors. PMID- 3038647 TI - Increased survival of skin flaps by scavengers of superoxide radical. AB - Elevation of rat abdominal skin flaps, followed by ligation and division of the left inferior neurovascular pedicle, resulted in only a 40% survival of the area normally perfused by the ligated artery and vein. Superoxide dismutase (SOD) (EC 1.15.1.1) administered i.v. (20,000 U/kg) 30 min before flap elevation increased survival to 52%. SOD derivatized with polyethylene glycol, which increases circulating half-life, was more effective, increasing survival to 80%. This protective effect resulted from the catalytic activity of the derivatized enzyme because inactivation by treatment with H2O2 eliminated its effect on skin flap survival. An equimolar mixture of Desferal and MnCl2, which catalyzes the dismutation of O2- in vitro, improved survival to 72%. Desferal-Fe3+, which lacks in vitro SOD activity, or Mn2+ alone did not affect the survival of skin flaps, but Desferal alone was nearly as effective as the Desferal-Mn2+ mixture. This effect of Desferal may result from acquisition of and subsequent removal of iron in vivo. These results support the view that the superoxide radical or a product derived from it plays a role in limiting the survival of island skin flaps. PMID- 3038648 TI - [Incidence and clinical significance of involvement of the right ventricle in acute inferior myocardial infarction]. AB - The purpose of our study was to evaluate, with noninvasive procedures, the incidence and the clinical picture of right ventricular involvement in patients with acute transmural inferior myocardial infarction. Our study group was constituted of 107 consecutive patients admitted to our Coronary Care Unit within 10 hours from the onset of symptoms; in every patient a standard 12-leads ECG and the precordial leads V3R and V4R were obtained at admission in CCU and then every 12 hours. 80 patients underwent B-mode echocardiographic evaluation within 36 hours and in 93 patients a myocardial scintigraphy was performed, between the 48th and 72nd hour from the onset of chest pain, 1-2 hours after injection of Tc 99m-pyrophosphate. RESULTS: 45 patients (42.1%) had ECG positive for right ventricular infarction, 49 patients (51.6%) had positive Tc-99m-pyrophosphate scintigraphy and 24 patients (30%) positive echocardiography. By using the positivity of ECG and another method at least, patients were separated into 2 groups: group A (associated inferior and right infarction) was constituted of 45 patients, and group B (isolated inferior infarction) was constituted of 62 patients. In group A we noted a higher incidence of hypotension (systolic blood pressure less than 100 mmHg) and oliguria (less than 30 ml/h)- p less than 0.01-, of 2nd and 3rd A-V blocks-p less than 0.001- and primary ventricular fibrillation -p less than 0.01. The incidence of parossistic atrial fibrillation, severe bradycardia or SA blocks and of mortality was not statistically different between the two groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3038649 TI - The action of trimebutine maleate on gastrointestinal motility is mediated by opiate receptors in human subjects. AB - The effects of trimebutine on interdigestive motor activity of the small intestine and the colon were studied in healthy volunteers. Trimebutine induced in all subjects phase III-like motor complexes, which were similar to spontaneously occurring phase III complexes of the interdigestive cycle in antrum and duodenum. Trimebutine-induced complexes were suppressed by naloxone. Trimebutine and/or naloxone had no consistent effect on interdigestive contractile activity in the colon. PMID- 3038651 TI - Involvement of enkephalins and other endogenous opioids in the regulation of esophageal motility. PMID- 3038650 TI - Brain, spinal cord and peripheral sites of action of enkephalins and other endogenous opioids on gastrointestinal motility. PMID- 3038652 TI - Endogenous opioids, gastric motility, and gastric emptying. PMID- 3038653 TI - Enkephalins, other endogenous opioids and colonic motility in dog and man. PMID- 3038654 TI - [Opioid peptides: metabolism and receptors]. PMID- 3038656 TI - [Effect of trimebutine on cholinergic transmission in neurons of the inferior mesenteric ganglion of the rabbit]. AB - We analyzed the effects of trimebutine on the synaptic activity of neurons of the rabbit inferior mesenteric ganglion, using intracellular recording techniques. The synaptic activity was produced by subthreshold stimuli (0.5 Hz) applied individually, on lumbar splanchnic and lumbar colonic nerves. These stimuli triggered cholinergic responses corresponding to fast excitatory postsynaptic potentials. In 8 of 20 neurones tested trimebutine (10(-6) g/ml) produced an inhibition of excitatory postsynaptic potentials, without any change in the resting membrane potential. In 6 of 20 neurons tested, trimebutine produced, successively, an early facilitation followed by a late inhibition of excitatory postsynaptic potentials. Both effects occurred without change in the resting membrane potential. The inhibitory and facilitatory effects of trimebutine were accompanied, by an increase and a decrease in the number of failures of nerve stimulation respectively. These results indicate that inhibitory and facilitatory effects of trimebutine correspond respectively to a decrease and an increase in the amount of acetylcholine released from presynaptic nerve terminals originating from the spinal cord and the distal colon. PMID- 3038655 TI - [Involvement of opiate receptors in the mode of action of trimebutine]. AB - Several studies in dogs, cats, rabbits and humans have suggested that the motility stimulating properties of trimebutine (TMB) are mediated by peripheral opiate receptors. The present work deals with the capacity of TMB and its N desmethyl metabolite (NDTMB) to displace mu, delta and kappa specific ligands from their receptors using guinea-pig brain membranes and ileal myenteric plexus synaptosomal membrane preparations. The activity of both compounds on the twitch response induced by transmural stimulation on the guinea-pig ileum as well as the mouse and rabbit vas deferens was also investigated. These preparations have been proposed to be specific for the mu, delta and kappa receptor subtypes respectively. TMB (0.2 to 1.8 microM) and NDTMB (0.3 to 6 microM) displayed a good affinity for all receptor subtypes in brain and myenteric plexus preparations. Both compounds also inhibited the twitch response of all three isolated organ preparations. The decreasing order of IC50's of TMB ranged from 0.75 microM in the guinea-pig ileum to 7.1 and 39 microM in the vas deferens of the rabbit and the mouse respectively. These results indicate that TMB and NDTMB possess mu, delta as well as kappa agonist properties without true specificity for one or the other of these subtypes. They also confirm that activation of peripheral mu, delta and kappa opiate receptors mediate the gastrointestinal motility effect of TMB. PMID- 3038657 TI - Local actions of trimebutine on canine gastrointestinal tract. AB - The local actions of trimebutine on the circular muscle of canine gastrointestinal tract were studied after close intraarterial injection. The effects resembled those of metenkephalin at all sites. In stomach, trimebutine had no excitatory effects, but inhibited responses mediated by cholinergic post ganglionic nerves. In small intestine, trimebutine stimulated the quiet gut by probably both neural and direct smooth muscle mechanisms, and it inhibited the field-stimulated phasic contractions. In large intestine, trimebutine had no excitatory actions and only weak inhibitory actions on the field-stimulated gut. Excitatory actions most likely seem to use the mu or delta receptors while inhibitory actions may focus on kappa opiate receptors. PMID- 3038658 TI - [Hepatocellular carcinoma and autoimmune chronic active hepatitis]. PMID- 3038659 TI - [Absence of specific IGM of cytomegalovirus, Epstein-Barr virus and hepatitis B virus in patients with primary biliary cirrhosis]. PMID- 3038660 TI - Occult radiopaque jaw lesions in familial adenomatous polyposis coli and hereditary nonpolyposis colorectal cancer. AB - The purposes of this study were to determine the association, in 10 pedigrees, between adenomatous polyposis coli, hereditary nonpolyposis colorectal cancer, and occult radiopaque jaw lesions, and to assess whether these radiodensities are predictors for adenomatous polyposis. In seven kindreds with adenomatous polyposis, all patients with polyps had jaw lesions; in one kindred, no jaw lesions were found. In one of two kindreds with hereditary nonpolyposis colorectal cancer, no affected individuals had jaw lesions. In the other, the 1 affected patient with dental radiographs had generalized jaw lesions. Twelve children less than 16 yr old at risk for adenomatous polyposis were observed. Seven children with jaw lesions developed polyps after a mean interval of 4 yr. Five children without jaw lesions were polyp-free during a 5-10-yr follow-up. Thus, occult jaw lesions are consistently found only in some families with adenomatous polyposis coli, providing support for heterogeneity in polyposis syndromes. Jaw lesions are good predictors for polyp development in kindreds with adenomatous polyposis coli and jaw lesions. Their role as markers in hereditary nonpolyposis colorectal cancer needs exploration. PMID- 3038661 TI - Portal venous hemodynamics in hepatocellular carcinoma. Effects of hepatic artery embolization. AB - Portal hemodynamics were studied in 55 patients with hepatocellular carcinoma in comparison with 41 normal subjects, using the duplex system that consists of an electronic sector scanner and a pulsed Doppler velocitometer. Changes of portal hemodynamics after transcatheter hepatic artery embolization were also investigated in 15 of the patients with hepatocellular carcinoma. The duplex system showed that 9 of the 55 had no Doppler signal in the portal trunk, suggesting portal vein thrombosis, 2 had hepatofugal flow in the portal trunk indicative of arterioportal shunts, and 44 had hepatopetal flow in the portal trunk. One of the 9 patients with no significant portal venous flow showed hepatopetal flow in collateral veins at the porta hepatis, suggesting cavernous transformation of the portal vein. All of these ultrasound findings were confirmed by subsequent celiac-mesenteric angiography. In 44 of the 55 patients there was no tumor invasion in the portal trunk, and portal venous flow was found to be close to that of normal subjects regardless of the stage or size of tumor, and tumor invasion into relatively large portal branches. After transcatheter hepatic artery embolization, portal venous flow was increased, even on the next day, and it remained increased for at least 2 wk. Thus, the duplex system is useful to study qualitative and quantitative changes of portal hemodynamics in hepatocellular carcinoma. Our observations suggest that the portal venous flow is kept relatively constant by some homeostatic mechanism even in advanced hepatocellular carcinoma until the tumor invades into the portal trunk, and that it increases when hepatic arterial flow is occluded. PMID- 3038662 TI - New solubilizing mixture for pigment gallstones. PMID- 3038663 TI - [Receptors for thrombin on lymphocytes and their subpopulations]. PMID- 3038664 TI - Human placental lipid peroxidation--II. NADPH and iron dependent stimulation of microsomal lipid peroxidation by paraquat. AB - Paraquat, a widely used herbicide, was found to cause a marked stimulation of lipid peroxidation in the human placental microsomes in vitro. Both NADPH and chelated iron were necessary to observe paraquat-stimulated lipid peroxidation. The malondialdehyde accumulation in the incubation medium increased with increase in time, protein and paraquat concentration. The reaction did not exhibit the initial lag phase suggesting that endogenous membrane-bound antioxidants in human placental microsomes are either absent or present in extremely small quantities. PMID- 3038665 TI - Action of xanthine-xanthine oxidase system on microsomal benzo(a)pyrene metabolism in vitro. AB - The effect of superoxide anion-radical and other reactive oxygen species on the metabolism of benzo(a)pyrene was studied with isolated mouse liver microsomes. Reactive oxygen species were generated in vitro by xanthine-xanthine oxidase plus Fe3+ X FeEDTA and benzo(a)pyrene metabolism was followed by reverse-phase high pressure liquid chromatography. The following results were obtained: The reactive oxygen species induced one-electron oxidation of benzo(a)pyrene and increased production of free epoxide as well as protein-binding intermediates. The reactive oxygen species triggered microsomal lipid peroxidation in the presence of Fe3+ X FeEDTA. As a result of microsomal lipid peroxidation a decreased activity of cytochrome P-450, epoxide hydrolase and UDP-glucuronyltransferase was found. It is suggested that active oxygen species changed the balance between bioactivation and conjugation of benzo(a)pyrene metabolites causing accumulation of the epoxide and protein-binding intermediates. The role of iron ions and chelates in this process is discussed. PMID- 3038666 TI - Age-related changes in brain biogenic monoamines and monoamine oxidase. AB - In experiments on 2-, 10- and 22-month old rats, it was found that essential age related changes occurred in the brain level of biogenic monoamines (BMA) and in the monoamine oxidase (MAO) activity. In 22-month old rats the levels of dopamine (DA), noradrenaline (NA) and serotonin (5-HT) markedly declined in most of the brain structures studied. 5-HT significantly decreased in the frontal cortex, striatum and hypothalamus. DA decreased in the cerebral cortex and striatum and NA in the cerebral cortex. However, the NA level in the striatum of 22-month old rats was increased as compared to that in 2-month old rats. In most cases we observed significant differences (a decrease mainly) also in the level of BMA in 22-month old rats as compared to 10-month old rats. The differences, if any, in the BMA levels between 10- and 2-month old rats were less pronounced. The level of 5-hydroxyindole acetic acid (5-HIAA) in the cerebral cortex and striatum of 22 month old rats was significantly higher as compared to that in 2-month old rats. The MAO-T and MAO-A activities in the brain structures studied were significantly higher in 22-month old rats as compared to those in 2-month old rats. The possibility that the age-related changes in brain neurotransmission might be an important element in the neurochemical basis of some behavioral changes in advanced age is considered. PMID- 3038667 TI - The effects of parenteral injections of adenosine and its analogs on blood pressure and heart rate in the rat. AB - The dose-response effects of adenosine and its analogs on cardiovascular parameters were examined in rats following intravenous administration. 5'-N ethylcarboxamidoadenosine (NECA) was by far the most potent analog in reducing mean arterial blood (PA) pressure while N6-(3-pentyl)-adenosine exerted the most potent bradycardic action. The N6-substituted (S)-diastereoisomers were substantially less potent in reducing PA and heart rate than NECA and the N6 substituted (R)-diastereoisomers. The cardiovascular effects of adenosine analogs persist, to varying degrees, much longer than those of adenosine itself. PMID- 3038668 TI - Mechanisms involved in catecholamine effect on glycogenolysis in catfish isolated hepatocytes. AB - Isolated catfish hepatocytes were treated with epinephrine, norepinephrine, isoproterenol, and phenylephrine in the presence or in the absence of propranolol or phentolamine as beta and alpha inhibitors, respectively. Glycogen phosphorylase a activity and glycogen content, as well as glucose released from cells, were tested. Phosphorylase activity was stimulated by all the catecholamines and was accompanied by a decrease of glycogen content in cells and by an increase in glucose output into the medium. Whereas phentolamine did not affect the catecholamine action on any parameter considered, propranolol inhibited the effect of epinephrine, norepinephrine, and phenylephrine, but hardly altered that of isoproterenol. The effect of epinephrine and norepinephrine, as modified by propranolol and not by phentolamine, is consistent with a beta action of these catecholamines. The fact that propranolol and not phentolamine inhibited the phenylephrine effect indicates that in catfish hepatocytes phenylephrine behaves as a beta agonist and/or that propranolol may also bind to alpha receptors. Results also indicate that in catfish liver cells isoproterenol, whose effect is scarcely influenced by propranolol, is not a pure beta agonist. PMID- 3038669 TI - A new generalizable test for detection of mutations affecting Tn10 transposition. AB - We describe here a new rapid screen that allows easy detection of transposon or host mutations that affect Tn10 transposition in Escherichia coli. This test involves a new Tn10 derivative called the "mini-lacZ-kanR fusion hopper" or mini Tn10-LK for short. This element does not direct expression of beta-galactosidase when present at its original starting location on a suitably engineered plasmid or phage genome because it lacks appropriate transcription and translation start signals. However, transposition of this element into the chromosome of E. coli lacZ- bacteria leads to productive fusions in which the lacZ gene within the transposon is expressed from external chromosomal signals. Such fusions are readily detectable on MacConkey lactose indicator plates as red (Lac+) papillae inside of white (LacZ-) colonies. The length of time required to see red papillae appearing in a white colony sensitively and accurately reflects the transposition frequency of the mini-transposon within the colonies. Differences in times for color formation are sensitive enough that 10-fold differences in transposition frequency can readily be detected. This papillation assay can be used to identify mutant clones in which the frequency of Tn10 transposition is either increased or decreased. We have successfully used the assay to identify mutations in the terminal sequences of Tn10; mutations in the Tn10 transposase gene or the bacterial host can be isolated just as easily. This screen should be readily adaptable to transposable elements other than Tn10. PMID- 3038670 TI - A Tn10-lacZ-kanR-URA3 gene fusion transposon for insertion mutagenesis and fusion analysis of yeast and bacterial genes. AB - We describe here a new variant of transposon Tn10 especially adapted for transposon analysis of cloned yeast genes; it can equally well be used for analysis of prokaryotic genes. We have applied this element to analysis of the LEU2, RAD50, and CDC48 genes of Saccharomyces cerevisiae. This transposon, nicknamed mini-Tn10-LUK, contains a lacZ gene without efficient transcription or translation start signals, an intact URA3 gene, and a kanR determinant. The lacZ gene can be activated by appropriate insertion of the element into an actively expressed gene. Other yeast genes can easily be substituted for URA3 in the available constructs. The mini-Tn10-LUK system has several important advantages. Transposition events occur in Escherichia coli at high frequency and into many different sites in yeast DNA. It is easy to obtain enough insertions to sensitively define the functional limits of a gene. Transposon insertions can be obtained in a single step by standard transposon procedures and can be screened immediately for phenotype either in yeast or in E. coli. The LacZ phenotypes of the insertion mutations provide a good circumstantial indication of the orientation of the target gene. Under favorable circumstances, usable lacZ protein fusions are created. Transposon insertion mutations obtained by this method directly facilitate additional genetic, functional, physical and DNA sequence analysis of the gene or region of interest. PMID- 3038671 TI - Temporal and spatial heterogeneity of mtDNA polymorphisms in natural populations of Drosophila mercatorum. AB - Restriction endonuclease analysis of mtDNA was used to examine the genetic relatedness of several geographically separated isolines of the Drosophila mercatorum subgroup. In addition, we examined the temporal and spatial distribution of two mtDNA restriction site polymorphisms produced by the enzymes BstEII and BstNI at a single locality--Kamuela, Hawaii. Due to small sample sizes of some collections and the undesirable dependance of the estimation of polymorphism frequency on its variance, an arcsin square root transformation of the frequency data was used. We also use an Fst estimator of our transformed frequencies to demonstrate considerable spatial and temporal differentiation within the Kamuela population. In contrast, isozyme data from the same population reveals no pattern of differentiation. The temporal and geographic heterogeneity and population subdivision detected with mtDNA analysis also is consistent with the known dispersal behavior and ecological constraints of this species. The mtDNA data in conjunction with the isozyme data show that the population structure of the Kamuela D. mercatorum is close to the boundary line separating panmixia from subdivision, a conclusion that could not be made from isozyme data alone. PMID- 3038672 TI - Evolution of the ribosomal DNA spacers of Drosophila melanogaster: different patterns of variation on X and Y chromosomes. AB - Length variation of the ribosomal gene spacers of Drosophila melanogaster was studied. Analysis of 47 X chromosomal and 47 Y chromosomal linked rDNA arrays collected from five continents indicates that the arrays on the two chromosomes differ qualitatively. The Y-linked arrays from around the world share little or no similarity for either their overall length or the organization of their spacers. Most of the X-linked arrays do, however, share a major length spacer of 5.1 kb. In addition, those X-linked arrays that have a major 5.1-kb band have similar spacer organization as demonstrated by genomic DNA digestions with several restriction enzymes. These data strongly support the hypothesis that spacer length patterns on only X-linked genes are maintained primarily by natural selection. PMID- 3038674 TI - Organ-specific expression of maize Adh1 is altered after a Mu transposon insertion. AB - A new, unstable, organ-specific Adh1 mutant was isolated from a Robertson's mutator line by germinating kernels under partial anaerobic conditions. Families of kernels which showed segregation of a conditional anaerobic lethal phenotype were identified. One mutant, Adh1-3F1124, was shown to express approximately 6% normal levels of ADH1 in seed and anaerobically treated seedlings but expresses normal levels of ADH1 in pollen, the male gametophyte. The ADH1 polypeptide encoded by the mutant allele was found to be indistinguishable from that encoded by the Adh1-3F progenitor but its message levels were lower in seed and seedlings. Robertson's mutator lines are known to carry Mu transposons that cause increased mutation rates. Genomic Southern analysis of Adh1-3F1124 and Adh1-3F showed the presence of a 1.85 kbp insertion at the 5' region of Adh1. Comparison of the DNA sequences revealed that a Mu 1-like element was inserted 31 bp 5' from the transcriptional start site of Adh1-3F1124 gene. The insertion of the Mu element creates an additional TATA box by duplicating the 9 bp genomic sequence- ATATAAATC--at the site of insertion. Consequently, there are two potentially functional TATA sequences, separated by the 1.85 kbp Mu element, 5' to the transcriptional start site. It is not yet understood how such an arrangement alters the organ-specific expression of Adh1. PMID- 3038673 TI - Physical analysis of Tn10- and IS10-promoted transpositions and rearrangements. AB - We have investigated by Southern blot hybridization the rate of IS10 transposition and other Tn10/IS10-promoted rearrangements in Escherichia coli and Salmonella strains bearing single chromosomal insertions of Tn10 or a related Tn10 derivative. We present evidence for three primary conclusions. First, the rate of IS10 transposition is approximately 10(-4) per cell per bacterial generation when overnight cultures are grown and plated on minimal media and is at least ten times more frequent than any other Tn10/IS10-promoted DNA alteration. Second, all of the chromosomal rearrangements observed can be accounted for by two previously characterized Tn10-promoted rearrangements: deletion/inversions and deletions. Together these rearrangements occur at about 10% the rate of IS10 transposition. Third, the data suggest that intramolecular Tn10-promoted rearrangements preferentially use nearby target sites, while the target sites for IS10 transposition events are scattered randomly around the chromosome. PMID- 3038675 TI - Molecular cloning of DNA complementary to Drosophila melanogaster alpha-amylase mRNA. AB - Several lambda clones containing cDNAs from Drosophila melanogaster were identified in a lambda cDNA bank using two different approaches: (i) cross species hybridization using a mouse amylase cDNA probe, and (ii) probing with a differential probe, generated from Drosophila RNA. An amylase cDNA fragment was used, in turn, for the isolation and characterization of amylase genomic clones. The size of the Drosophila amylase mRNA was estimated at 1650 b. This is comparable with the size of the murine amylase messenger that encodes a protein of similar molecular weight. In Drosophila larvae, amylase mRNA can account for as little as 0.01% of the poly(A)+ RNA under conditions of dietary glucose repression or greater than 1% of poly(A)+ RNA under derepressing dietary conditions. PMID- 3038676 TI - Physical characterization of katG, encoding catalase HPI of Escherichia coli. AB - The gene encoding the bifunctional catalase-peroxidase HPI from Escherichia coli was located on a 3.8-kb HindIII fragment of the Clarke and Carbon plasmid pLC36 19 using transposon Tn5 insertions. This fragment was subcloned into the HindIII site of pAT153 to create pBT22. The size of the insert was reduced by BAL 31 digestion of one end to an apparent minimum size for catalase expression of approx. 2.5 kb as determined by complementation and expression in maxicell strains. Further reduction in size or digestion from the opposite end inactivated the gene. The location and orientation of the promoter at the 0 kb end of the insert in pBT22 was confirmed by cloning a 320-bp BglII fragment into the promoter-cloning vector pKK232-8. Differences in the Southern blots of genomic DNA from a wild-type strain and a katG17::Tn10 mutant digested with HincII and probed with pBT22 confirmed that the transposon previously mapped in katG was located in the 2.5-kb coding region for HPI. PMID- 3038677 TI - Nucleotide sequence of the DNA polymerase gene of herpes simplex virus type 2 and comparison with the type 1 counterpart. AB - The complete nucleotide sequence of the DNA polymerase gene of herpes simplex virus (HSV) type 2 strain 186 has been determined. The gene included a 3720-bp major open reading frame capable of encoding 1240 amino acids. The predicted primary translation product had an Mr of 137,354, which was slightly larger than its HSV-1 counterpart. A comparison of the predicted functional amino acid sequences of the HSV-1 and HSV-2 DNA polymerases revealed 95.5% overall amino acid homology, the value of which was the highest among those of the other known polypeptides encoded by HSV-1 and HSV-2. The functional amino acid changes were spread in the N-terminal one-third of the protein, whereas the C-terminal two third was almost identical between the two types except a particular hydrophilic region. A highly conserved sequence of 6 aa, YGDTDS, which has been observed in DNA polymerases of HSV-1, Epstein-Barr virus, adenovirus, and vaccinia virus, was also present at positions 889 to 894 in the C-terminal region of HSV-2 DNA polymerase. PMID- 3038678 TI - Specific association between type-II intracisternal A-particle elements and other repetitive sequences in the mouse genome. AB - The majority of type-II intracisternal A-particle (IAP) element clones isolated from a mouse genomic library also contained highly repetitive DNA sequences in addition to the moderately repetitive IAP elements. Further analysis revealed that eleven of the twelve clones contained sequences of the mouse L1 family. One clone contained four copies of a limited region of the 3' end of the L1 element in a 12-kb stretch of sequence. This clone also contained a newly identified repetitive sequence which is found associated with type-II IAP elements. Type-II IAP elements were completely methylated in mouse embryo DNA; in myeloma cells, partial demethylation of the sequences correlated with known transcriptional activity of the IAP subclasses. Analysis of genomic DNA showed that association with other repetitive sequences appears to be a general property of many type-II IAP elements and may reflect their location in a particular chromosomal environment. PMID- 3038679 TI - Interposon mutagenesis of soil and water bacteria: a family of DNA fragments designed for in vitro insertional mutagenesis of gram-negative bacteria. AB - We have constructed a series of derivatives of the omega interposon [Prentki and Krisch, Gene 29 (1984) 303-313] that can be used for in vitro insertional mutagenesis. Each of these DNA fragments carries a different antibiotic or Hg2+ resistance gene (ApR, CmR, TcR, KmR or HgR) which is flanked, in inverted orientation, by transcription and translation termination signals and by synthetic polylinkers. The DNA of these interposons can be easily purified and then inserted, by in vitro ligation, into a plasmid linearized either at random by DNase I or at specific sites by restriction enzymes. Plasmid molecules which contain an interposon insertion can be identified by expression of its drug resistance. The position of the interposon can be precisely mapped by the restriction sites in the flanking polylinker. To verify their properties we have used these omega derivatives to mutagenize a broad host range plasmid which contains the entire meta-cleavage pathway of the toluene degradation plasmid pWW0 of Pseudomonas putida. Insertion of these interposons in the plasmid between the promoter and the catechol 2,3-dioxygenase (C23O) gene dramatically reduced the expression of this enzyme in Escherichia coli. We also show that when a plasmid containing an omega interposon is transferred by conjugal mobilization from E. coli to P. putida, Agrobacterium tumefaciens, Erwinia chrysanthemi, Paracoccus denitrificans or Rhizobium leguminosarum, the appropriate interposon drug resistance is usually expressed and, compared to the non-mutated plasmid, much reduced levels of C23O activity are detected. Thus, the selection and/or characterization of omega insertional mutations can be carried out in these bacterial species. PMID- 3038680 TI - The first-step transfer-DNA injection-stop signal of bacteriophage T5. AB - Bacteriophage T5 is different from most phages in that its DNA is injected in two steps during infection. The region containing the injection stop signal (iss) has been cloned and sequenced and found to contain numerous large repeats and inverted repeats which may be part of the iss. The most impressive of these are the 31-bp repeat units (rb) which are present three times in 99 bp. The rb repeats, themselves, contain inverted repeats so that mutually exclusive stem-and loop structures may potentially form, not only within the repeats, but also between them. Another pair of repeats (21 bp each) contains two sequences resembling DnaA protein-binding sites. The region sequenced also contains one of the T5 site-specific strand interruptions and this was found to lie at the base of a perfect 9-bp palindrome. PMID- 3038681 TI - In vivo selection and characterization of internal deletions in the lamB::lacZ gene fusion. AB - Strain Pop3299 contains the lamB::lacZ42-12 gene fusion that encodes a hybrid protein that is efficiently exported to the cellular envelope of Escherichia coli. As a result of this efficient export, this strain is killed by the inducer maltose and unable to grow on minimal media supplemented with lactose. In an attempt to isolate mutants in which export of the hybrid protein is altered, we selected Lac+ mutants of strain Pop3299 on lactose tetrazolium media. Unlike mutants previously isolated on lactose minimal media, all the mutants we obtained carried large deletions within the lamB::lacZ gene fusion. Thus, it appears that the type of selection employed affects the type of mutations obtained. We have analyzed the nucleotide sequences of representative mutants, and demonstrate a correlation between the deletion size and the export-related maltose and lactose phenotypes. In addition, we demonstrate that the deletions do not appear to arise from regions of micro-homology. PMID- 3038682 TI - Cloning in Escherichia coli of the gene specifying the DNA-entry nuclease of Bacillus subtilis. AB - With the aim of cloning genes involved in transformation of Bacillus subtilis, a set of transformation-deficient mutants was isolated by means of insertional mutagenesis with plasmid pHV60 (Vosman et al., 1986). Analysis of these mutants showed that those mapping in the aroI region lacked the DNA-entry nuclease activity. Plasmid pHV60 derivatives, containing flanking chromosomal DNA fragments, were isolated from these mutants and were used to screen a library of B. subtilis chromosomal DNA in phage lambda EMBL4. In Escherichia coli lysates, prepared with the phages that hybridized to the pHV60-based probe, a prominent nuclease activity could be detected. The nuclease encoded by the phage DNA had the same Mr as the B. subtilis DNA-entry nuclease and its activity was strongly stimulated by Mn2+, which is also characteristic for the B. subtilis DNA-entry nuclease. From these results it was concluded that the gene specifying the B. subtilis DNA-entry nuclease had been cloned. It was shown that the nuclease activity was specified by a 700-bp EcoRI-PstI fragment. PMID- 3038683 TI - A family of lambda phage cDNA cloning vectors, lambda SWAJ, allowing the amplification of RNA sequences. AB - This paper describes the construction and characterization of a family of lambda phage cDNA cloning vectors that allows high-efficiency directional cDNA cloning and selective amplification of either sense or antisense cRNA sequences. These vectors contain several unique restriction sites (EcoRI, XbaI, and SacI) positioned between two specific phage promoters, SP6 and T7. This system facilitates the in vitro preparation of single-stranded (ss) RNA molecules that should be useful in subtractive hybridization and in situ hybridization procedures. Using subtractive hybridization and this vector system, it should be possible to identify sequences present in one cDNA library and not another. In addition, it should be possible to construct subtracted cDNA libraries in these vectors and to generate high specific activity, ss, antisense cRNA probes directly from DNA prepared from the whole subtracted library or from individual clones. PMID- 3038684 TI - Role of the merT and merP gene products of transposon Tn501 in the induction and expression of resistance to mercuric ions. AB - The bacterial transposon Tn501 carries, in addition to the genetic information for its own transposition, the genes of the mer operon (in the order merRTPAD), which code for resistance to Hg2+ ions. The basis for the resistance to Hg2+ is the enzymatic reduction of Hg2+ to Hg0 by mercuric reductase, the product of the Tn501 merA gene. We show here that deletion of the merT and merP genes from Tn501 leads to almost complete loss of the resistance phenotype, even if mercuric reductase is still present in the cytoplasm. Expression of the merT and merP genes in the absence of mercuric reductase gives a mercury-supersensitive phenotype. Mercury-dependent induction of transcription of the mer operon can occur in the absence of the merT and merP gene products. However, this induction is reduced by the presence of mercuric reductase in the cell. We conclude that one or both of the merT and merP genes of Tn501 are required for the expression of the mercury-resistance phenotype. They may in addition have a role in maximising the induction of the mer operon in the presence of Hg2+ ions. This is consistent with their proposed role in transport of Hg2+ into the cytoplasm. PMID- 3038685 TI - Production of chimeric protein coded by the fused viral H-ras and human N-ras genes in Escherichia coli. AB - A new method for the production of a chimeric protein of two related genes has been developed. The nucleotide sequences of the region from the N terminus to the 86th amino acid (aa) residue of human N-ras and of the Harvey sarcoma virus (Ha MuSV) H-ras are 80% homologous. We isolated the DNA fragment encoding the N terminal portion up to the 70th aa residue from plasmid pH-1 which encodes the total genome of Ha-MuSV, and the DNA fragment encoding the C-terminal portion from the 40th aa to the C terminus from plasmid p6a1 which includes the human N ras cDNA but lacks the N-terminal portion. After partial digestion of both fragments with phage lambda exonuclease, which creates 3'-protruding ends, a hybrid was formed between 73% homologous single-stranded DNA portions at the 3' ends of both fragments. The hybrid was recloned on pBR322 after repairing with Escherichia coli DNA polymerase I and DNA ligase. The chimeric v-H/N-ras gene composed of the N-terminal portion of v-H-ras gene and the remaining region of N ras gene was inserted into an expression vector containing two tandem trp promoters and a terminator, and expressed in E. coli. The chimeric protein was found to accumulate to approx. 10% of total cellular proteins. PMID- 3038686 TI - A family of yeast expression vectors containing the phage f1 intergenic region. AB - The construction and characterization of a family of yeast expression vectors is described. They have the following features: plasmid replication and selection (ApR) in Escherichia coli, packaging of single-stranded (ss) DNA upon infection of E. coli with a filamentous helper phage, replication in Saccharomyces cerevisiae based on the 2 mu plasmid origin of replication (ori), selection in yeast by complementation of LEU2 (pVT-L series, size 6.3 kb) or URA3 gene (pVT-U series, size 6.9 kb) and seven unique restriction sites for cloning within an 'expression cassette' which includes the promoter and 3' sequence of the ADH1 gene. The multiple cloning site as well as the ori and intergenic region of the phage f1 have been cloned in two orientations for convenient gene cloning and ssDNA strand selection. As a result any of these eight vectors can be chosen for cloning, expressing genes in yeast, sequencing and mutagenesis without the need for recloning into specialized vectors. PMID- 3038687 TI - Reverse-transcribed pseudogenes of U1 small nuclear RNA presumably amplified in the rat genome together with the flanking region. AB - We have examined 25 independent rat genomic clones each of which contains a U1 RNA gene or a pseudogene. We have found five clones whose restriction maps are identical with or overlapping one another. These clones contain sequences which are co-linear with that of U1 RNA except for one or two nucleotides (nt). They also contain 21-23-nt poly(A) stretches immediately after U1 RNA homology. In addition, the U1 RNA-poly(A) regions are surrounded by the same direct repeat sequences. Therefore, they seem to be pseudogenes which have been generated by the reverse transcription of poly(A)-tailed U1 RNA followed by the insertion of the transcript into the genome. Furthermore, conservation of the sequences extends over at least 18 kb of the flanking sequences. This suggests a family of conserved reverse-transcribed pseudogenes, which implies amplification of an original pseudogene. It is also suggested that the target sequence without insertion of a U1 RNA sequence has been amplified. The mechanisms for the amplification and sequence conservation are discussed. PMID- 3038688 TI - A low-copy-number vector utilizing beta-galactosidase for the analysis of gene control elements. AB - A low-copy-number vector, pFZY1, with the multiple restriction site linker of M13mp18 inserted upstream from a promoterless beta-galactosidase (beta Gal) coding lacZ gene has been constructed to provide a convenient and accurate system to analyze regulatory elements in vivo. The plasmid contains the oriF replication origin without the par locus and is present in the cell in one to two copies per genome. It is retained in the host by the presence of ampicillin, and each inserted promoter yielded consistent values of beta Gal activity under all the conditions tested. A series of tetracycline resistance (TcR) promoter fragments and lac promoter fragments have been compared in pFZY1 and the high-copy-number pKO-vector series. The transcriptional activity measured for different fragments containing the same TcR promoter varied within a six-fold range among the several constructs tested. Regulation of the wild-type lac promoter and mutants in pFZY1 was similar to that observed for lac promoters in the chromosome while their regulation in pKO-1mp18 was significantly affected by the high copy number, as expected. PMID- 3038689 TI - Analyses of alpha/beta-type gliadin genes from diploid and hexaploid wheats. AB - The alpha/beta-gliadin genes isolated from both hexaploid wheat (cv. Yamhill) and the diploid A genome progenitor Triticum urartu had remarkably similar sequences and differ by only a few point mutations. Primer extension analysis indicated that the transcriptional start points for individual genes in the family cluster within a few nucleotides. Comparison of the promoter region of several alpha/beta gliadin and B-hordein genes reveals two conserved regions at about -130 and -250 bp. DNA from the hexaploid cultivars, Cheyenne and Chinese Spring, and the diploid progenitors T. urartu and Aegilops squarrosa was analysed by Southern blotting. Restriction fragment lengths of the alpha/beta-gliadin genes varied only slightly between the various wheats, although the overall copy number varied significantly. A region between approx. -1700 and -700 bp upstream from the TATA box was highly repeated in all three wheat genomes. For the hexaploid-derived gene, over 1700 bp of sequence upstream from the TATA box was determined, revealing an additional open reading frame between approx. -1550 and -1250 bp relative to the gliadin TATA box. Northern blot analysis indicated that RNA homologous to this repeated sequence family was present only in developing seed and accumulated to a maximum at late stages of maturation. PMID- 3038690 TI - Inducible expression vectors incorporating the Escherichia coli atpE translational initiation region. AB - New expression vectors were constructed for use in strains of Escherichia coli. Their most important feature is a polylinker system that facilitates the insertion of a gene in an optimal relationship to the highly efficient E. coli atpE translational initiation region (from nucleotide -50 to the start codon). Three ATG-containing restriction endonuclease sites can be used for the insertion of the 5' end of a gene at, or near to, its translational initiation codon. These sites may alternatively be used for the creation of a suitable translational start codon. Transcription is started by the bacteriophage lambda major promoters pR and pL in tandem and terminated by the bacteriophage fd terminator. Transcriptional initiation is very effectively repressed at 28-30 degrees C by the product of the bacteriophage lambda cIts857 gene, which is also present on the vectors. Full induction is achieved by shifting the incubation temperature to 42 degrees C. The combination of highly efficient transcriptional and translational signals on these vectors allowed high-level expression of sequences encoding human interferon beta and interleukin 2 and of the E. coli atpA, sucC and sucD genes. PMID- 3038691 TI - Cloning and heterologous expression of an insecticidal delta-endotoxin gene from Bacillus thuringiensis var. aizawai IC1 toxic to both lepidoptera and diptera. AB - Bacillus thuringiensis var. aizawai IC1 synthesises an insecticidal protein delta endotoxin (130-135 kDa) that is toxic to both lepidopteran and dipteran larvae, and cross-reacts immunologically with certain monospecific lepidopteran toxins. A 166-kb plasmid from this bacterium was found to hybridise with an intragenic probe derived from the clone B. thuringiensis var. sotto lepidopteran-specific delta-endotoxin gene. A strongly hybridising 5.2-kb SstI fragment from var. aizawai plasmid DNA was cloned in pUC18. After subcloning of this DNA in Escherichia coli, recombinants were obtained that synthesised large amounts of a 130-135-kDa protein. The protein was deposited in the cytoplasm as microscopically visible inclusion bodies and lysates of these cells were found to be toxic to both lepidopteran and dipteran larvae by comparison with controls. The structural basis for the dual specificity of this var. aizawai toxin is now amenable to further study. PMID- 3038692 TI - RNA polymerase makes important contacts upstream from base pair -49 at the Escherichia coli galactose operon P1 promoter. AB - A G:C to T:A transversion at bp position -19 in the gal operon promoter region relieves the dependence of galP1 promoter activity on the cAMP-CRP complex. Deletion analysis shows that expression from the promoter is decreased on replacement of the sequence between 49 and 54 bp upstream from the P1 start point. Moreover, protection experiments show that RNA polymerase interacts with this region in open complexes at P1. We propose that this contact is necessary for optimal P1 activity; point mutations in the gal promoter region can alter DNA flexibility and hence the strength of this contact; CRP factor activates P1 transcription by favouring formation of this contact; and the gal repressor blocks P1 activity by binding to this zone. PMID- 3038693 TI - Phosphoribosylpyrophosphate synthetase of Bacillus subtilis. Cloning, characterization and chromosomal mapping of the prs gene. AB - The gene (prs) encoding phosphoribosylpyrophosphate (PRPP) synthetase has been cloned from a library of Bacillus subtilis DNA by complementation of an Escherichia coli prs mutation. Flanking DNA sequences were pruned away by restriction endonuclease and exonuclease BAL 31 digestions, resulting in a DNA fragment of approx. 1.8 kb complementing the E. coli prs mutation. Minicell experiments revealed that this DNA fragment coded for a polypeptide, shown to be the PRPP synthetase subunit, with an Mr of approx. 40,000. B. subtilis strains harbouring the prs gene in a multicopy plasmid contained up to nine-fold increased PRPP synthetase activity. The prs gene was cloned in an integration vector and the resulting hybrid plasmid inserted into the B. subtilis chromosome by homologous recombination. The integration site was mapped by transduction and the gene order established as purA-guaA-prs-cysA. PMID- 3038694 TI - Lorist6, a cosmid vector with BamHI, NotI, ScaI and HindIII cloning sites and altered neomycin phosphotransferase gene expression. AB - Four new features have been included in a phage-lambda-replicon-based cosmid vector, Lorist6. A ScaI restriction site allows insertion of blunt-ended DNA, partially digested with AluI or any other restriction enzyme which creates blunt ends after cleavage. NotI and BamHI sites enable construction of libraries using double digestion with NotI + Sau3AI. The promoter of the neo gene has been substituted by a promoter derived from the tet gene of pBR322, increasing expression of the resistance gene product. A transcriptional terminator has been placed downstream from the neo gene to prevent transcription into insert DNAs. PMID- 3038695 TI - Cloning of the riboB locus of Aspergillus nidulans. AB - We have complemented the riboB2 mutation of Aspergillus nidulans by transformation with a plasmid library of wild-type (wt) sequences. We have isolated, by marker rescue from a riboB+ transformant, a plasmid that complements riboB2 efficiently. From this plasmid we have subcloned an A. nidulans sequence that complements riboB2 efficiently and that integrates by homologous recombination at a site closely linked to the riboB locus. We conclude that this sequence contains the wt riboB+ allele. PMID- 3038697 TI - Induction of v-mos and activated Ha-ras oncogene expression in quiescent NIH 3T3 cells causes intracellular alkalinisation and cell-cycle progression. AB - We have tested the effect of the expression of the v-mos oncogene, and the activated human Ha-ras oncogene and its proto-oncogene form on the intracellular pH and on cell cycle progression. The Ha-ras proto-oncogene and the oncogenes under the transcriptional control of the MMTV-LTR promoter were introduced into NIH 3T3 cells. The cells were synchronized at G0/G1 in low-serum medium and the transfected genes were induced by glucocorticoid hormone addition to the growth medium. Expression of the v-mos and the activated form of Ha-ras oncogenes mimics the effect of growth factors and activates the Na+/H+ antiporter. The oncogenes increase the internal pH and provoke initiation of S-phase. The proto-oncogene form of Ha-ras does not induce intracellular alkalinisation and has only a weak mitogenic effect. Our results suggest the oncogenic proteins p21ras and p37mos influence the mitogenic signal transduction pathways at a point prior to the activation of the Na+/H+ antiporter. PMID- 3038696 TI - Expression and secretion in yeast of a 400-kDa envelope glycoprotein derived from Epstein-Barr virus. AB - The major envelope glycoprotein (gp350) of Epstein-Barr virus has been expressed and secreted in the yeast Saccharomyces cerevisiae as a 400-kDa glycoprotein. This is the first example of the secretion of such a large, heavily glycosylated heterologous protein in yeast. Since gp350 proved highly toxic to S. cerevisiae, initial cellular growth required repression of the expression of gp350. Using temperature- or galactose-inducible promoters, cells could be grown and the expression of gp350 then induced. After induction, the glycoprotein accumulated both intracellularly as well as in the culture medium. Only the most heavily glycosylated form was secreted, suggesting a role for N-linked glycans in directing secretion. The extent of O-linked glycosylation of the yeast-derived protein was similar to that of the mature viral gp350. N-linked glycosylation varied slightly depending upon culture conditions and host strain used and was more extensive than that associated with the mature viral gp350. Although there is no evidence that more than a single mRNA for the glycoprotein was expressed from the recombinant plasmid, variously sized glycoproteins accumulated in yeast at early stages after induction, probably reflecting intermediates in glycosylation. The yeast-derived glycoproteins reacted with animal and human polyclonal antibodies to gp350 as well as with a neutralizing murine monoclonal antibody to gp350, suggesting that this glycoprotein retains several epitopes of the native glycoprotein. PMID- 3038698 TI - Selectable genes for transformation of the fungal plant pathogen Glomerella cingulata f. sp. phaseoli (Colletotrichum lindemuthianum). AB - Glomerella cingulata f. sp. phaseoli (Gcp) was transformed using either of two selectable markers: the amdS + gene of Aspergillus nidulans, which encodes acetamidase and permits growth on acetamide as the sole nitrogen source and the hygBR gene of Escherichia coli which encodes hygromycin B (Hy) phosphotransferase and permits growth in the presence of the antibiotic Hy. The amdS+ gene functioned in Gcp under control of A. nidulans regulatory signals and hygBR was expressed after fusion to a promoter from Cochliobolus heterostrophus, another filamentous ascomycete. Protoplasts to be transformed were generated with the digestive enzyme complex Novozym 234 and then were exposed to plasmid DNA in the presence of 10 mM CaCl2 and polyethylene glycol. Transformation occurred by integration of single or multiple copies of either the amdS+ or hygBR plasmid into the fungal genome. There was no evidence of autonomous plasmid replication. Transformants were mitotically stable on selective and nonselective media. However, transforming DNA in hygBR transformants was observed to occasionally rearrange during nonselective growth, resulting in fewer copies of the plasmid per genome. These transformants were capable of infecting bean (Phaseolus vulgaris), the Gcp host plant, and after recovery from infected tissue were found to have retained both the transforming DNA unrearranged in their genomes and the Hy resistance phenotype. All single-conidial cultures derived from both amdS+ and hygBR transformants had the transplanted phenotype, suggesting that transformants were homokaryons. PMID- 3038699 TI - [Transmission of viruses of aquatic microflora]. PMID- 3038700 TI - [Complex hygienic indices for evaluating the risk of soil pollution as a result of using superphosphate and potassium chloride]. PMID- 3038702 TI - [Retrospective histological analysis of cervical intraepithelial carcinogenesis in relation to human papilloma virus infection]. PMID- 3038701 TI - [Body reactivity of workers in contact with the chemical allergen toluene diisocyanate]. PMID- 3038703 TI - The role of neuroplastic events in the physiopathogenesis and course of functional neurologic disorders. PMID- 3038704 TI - The short metyrapone test in chronic headache patients. AB - The short, overnight, metyrapone test with plasma ACTH and beta-endorphin (beta EP) determination has been performed in patients with migraine, other forms of headache, and healthy volunteers. A rise in plasma ACTH and beta-EP after metyrapone occurred in all investigated subjects. The rate of increase varied greatly from one individual to the other, but no significant differences between the mean values of plasma ACTH and beta-EP in different groups of headache patients and volunteers were observed. Three months of nadolol therapy in migraine patients caused a significant increase in plasma ACTH and decrease in beta-EP concentrations. Findings support the view that migraine patients do not have a primary dysfunction of the hypothalamus-pituitary-adrenal (HPA) axis as demonstrated by methods currently in use. PMID- 3038707 TI - [Endogenous opioids]. PMID- 3038705 TI - Autonomic disorders in patients with peripheral neuropathies. PMID- 3038706 TI - Non-vascular action of calcium blockers in migraine: pupillopharmacological study. AB - A pupillopharmacological study has been carried out to evaluate the influence of two calcium blockers (CBs), flunarizine and nimodipine, on iris neurotransmission in migraine patients. Pretreatment with an oral dose of flunarizine or nimodipine markedly decreased the mydriasis induced by instillation of tyramine, a noradrenaline releaser. On the contrary, both drugs did not change the mydriasis by conjunctival phenylephrine, a postsynaptic adrenoceptor stimulant. In addition, nimodipine abolished the miosis caused by echothiophate iodide eye drops, a putative releaser of substance P from trigeminal sensory nerve endings. These findings suggest the idea that CBs treatment in migraine syndromes acts at presynaptic level by attenuating calcium-dependent release of monoaminergic and peptidergic neurotransmitters. PMID- 3038708 TI - Loss of circadian rhythms of fetal behaviour in a totally adrenalectomized pregnant woman. AB - Fetal heart rate and fetal movements were recorded over a 24-hour interval in a totally adrenalectomized pregnant woman at term. Cortisol, ACTH, unconjugated estriol and 17 beta-estradiol were contemporaneously measured every 2 h in the maternal peripheral plasma. Owing to the substitution therapy the patient showed a loss of circadian variations of all the hormones investigated. Moreover, the circadian rhythms of fetal heart rate and fetal movements, usually present at term, were no longer evident. We can thus suggest that the circadian variations of plasma maternal cortisol could affect the alternations of fetal behaviour. PMID- 3038709 TI - Thrombomodulin activity is found in tissue thromboplastin preparations from placenta and from lung but not from brain. AB - Substantial thrombomodulin activity could be detected in tissue thromboplastin preparations from placenta or from lung but not from brain. When the amount of these preparations was adjusted to contain 1 unit of tissue factor activity, up to 0.85 units of thrombomodulin activity could be measured, corresponding to the generation of 17 pmol/ml/min of activated protein C when 1.5 microM human protein C was activated by 20 nM human alpha-thrombin in the presence of 5 mM CaCl2. After treatment by phospholipase C, thrombomodulin activity was reduced in these samples. Addition of mixed brain procoagulant phospholipids partially restored thrombomodulin activity in the phospholipase C-treated samples. These results emphasize the role of phospholipids in the expression of optimal thrombomodulin activity in tissue thromboplastin preparations from placenta or from lung. PMID- 3038710 TI - Inhibition of fibrinolytic system by liquoid (polyanetholesulfonate). A contributing factor in the liquoid-induced renal cortical necrosis. AB - Liquoid induces microvascular thrombosis and renal cortical necrosis in experimental animals. We hypothesized that thrombosis and renal cortical necrosis may, at least in part, result from the inhibition of the fibrinolytic system by liquoid. Effects of liquoid on plasminogen activation by rat kidney, purified human tissue plasminogen activator (TPA), urokinase, streptokinase, and on the amidolytic activities of TPA, urokinase, and plasmin were studied using chromogenic substrates and clot lysis. Liquoid had a strong inhibitory effect on the fibrinolytic system in vivo and in vitro. The inhibition was most effective at the plasminogen activation level, with activation by streptokinase being most susceptible. The demonstrated stoichiometric binding between liquoid and plasminogen, and to a lesser degree the direct inactivation of plasminogen activators and plasmin, is probably responsible for the reduction of plasminogen activation in circulation and in the kidneys. PMID- 3038711 TI - [Radio-isotopic scan of the inguino-scrotal region in varicocele]. PMID- 3038712 TI - [Herpes simplex virus and Guillain-Barre syndrome in a child]. PMID- 3038713 TI - [The modern management of malignant germ cell tumor of the ovary]. PMID- 3038714 TI - [Specific inhibitory action of a novel antidepressant paroxetine on 5-HT uptake]. AB - Effect of a novel antidepressant, paroxetine, on the uptake of serotonin (5-HT), noradrenaline (NA) and dopamine (DA) as well as on various neuro-receptors were investigated in comparison with those of the tricyclic antidepressants amitriptyline, chlorimipramine and imipramine. Paroxetine showed a potent 5-HT uptake inhibitory action, giving the NA/5-HT ratio of 886 in comparison with the ratios of 1.7, 15 and 1.5 for amitriptyline, chlorimipramine and imipramine, respectively. On the other hand, paroxetine showed almost no inhibitory action on the binding of the [3H]-labeled ligands examined in this study [( 3H]quinuclidinyl benzilate, [3H]5-HT, [3H]ketanserine, [3H]pyrilamine, [3H]dihydroalprenolol, [3H]prazosin, [3H]clonidine and [3H]spiroperidol). In contrast, the tricyclic antidepressants showed inhibitory action on a number of bindings and also revealed comparatively high affinities especially for muscarine, histamine-1 and alpha 1-adrenaline receptors responsible for the side effects. From the above findings, it can be concluded that paroxetine has only a weak affinity for various neuro-receptors and inhibits specifically 5-HT uptake. PMID- 3038715 TI - [Centrally and peripherally induced anorectic actions of salmon calcitonin (sCT) in rats: separation of its novel derivative [Gly8]-sCT]. AB - It has been reported based on animal studies that salmon calcitonin (sCT), besides hypocalcemic action, exhibits a variety of pharmacological actions. The anorectic action has been observed to ensue not only by central administration but also by peripheral injection, indicating that in clinical use to induce hypocalcemia or for other therapeutic purposes, the anorectic action may develop as a side effect. While studying the anorectic effect of [Gly8]-sCT in rats, a derivative of sCT having rather stronger hypocalcemic potency than the mother molecule, it was found that on peripheral injection, the derivative practically lacks the anorectic effect. Thus, a pharmacological evaluation of the novel peptide was made. When injected subcutaneously in rats at a dose level of 1 U/kg, sCT and the derivative induced similar patterns of hypocalcemia in either of which hypocalcemia reached a peak between 1 and 3 hr after injection. No notable difference existed between the action of the two peptides. In rats which were trained to take the daily food need within 2 hr (17:00 approximately 19:00), an intracerebroventricular injection of 1 U/rat of either peptide 30 min before food presentation significantly reduced both food and water intake, causing a loss of body weight as compared with the control which received the injection of saline alone. By subcutaneous injection of 100 U/kg, sCT was also active to decrease food and water intake. The effect was found to last longer than 24 hr and to cause a marked loss in body weight. In contrast, such effects did not develop in rats treated with the derivative. Both peptides were able to suppress the specific binding of 125I-sCT to the membrane fraction of rat brain and kidney.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3038716 TI - Small hepatocellular carcinoma: an Italian experience. AB - Recent reports have demonstrated the possibility of using ultrasound to detect hepatocellular carcinomas under 3 cm (small HCC). Our aim was to assess the incidence of small HCC in an Italian ultrasonographic series. Among 17,133 scans of unselected patients, we detected 85 HCC, 9 of which were under 3 cm. All patients had cirrhosis and 6 were HBsAg positive. An echo-guided biopsy was performed in all cases, and the diagnosis was always correct. We conclude that echographic follow-up is warranted in Italy for cases at risk for HCC such as HBsAg+ cirrhotics and patients with chronic aggressive HBsAg+ hepatitis older than 35, in agreement with the reports of Far Eastern authors. PMID- 3038717 TI - Biliary immunoreactive trypsin in man. AB - Small, but significant amounts of immunoreactive trypsin/trypsinogen (IRT) are excreted in bile, obtained via percutaneous transhepatic catheter in patients with complete distal bile duct obstruction, and thus uncontaminated with pancreatic juice. The major serum proteolytic enzyme inhibitors alpha 2 macroglobulin and alpha 1-antitrypsin are also present in very small amounts in bile. The bile-to-serum ratios of these inhibitors are much lower (approximately two orders of magnitude) than that for IRT. The role of these inhibitors and their relative importance in biliary proteolytic enzyme inhibition is unknown. PMID- 3038718 TI - Morphological precursors of hepatocellular carcinoma: a morphometrical analysis. AB - Morphometrical analysis of different types of liver cells was performed in cases of chronic active hepatitis (CAH), liver cirrhosis (LC), and hepatocellular carcinoma (HCC). The area and a form factor (form AR) of nuclei and cytoplasms were determined, and the nucleo-cytoplasmic ratio (N/C) was calculated in normal hepatocytes, HBsAg-positive cells, large dysplastic cells, cancer cells and in a liver cell population identifiable as "small dysplastic cells" (small cells). The nucleo-cytoplasmic ratio of the small cells and of the neoplastic cells was roughly the same and the highest. Large dysplastic cells showed a small nucleo cytoplasmic ratio, almost comparable with that of normal hepatocytes and similar to that of HBsAg-positive cells. Since cellular precursors of liver cancer are expected to have a nucleo-cytoplasmic ratio similar to that of neoplastic cells, our morphometrical evaluation indicates small cells as the true precancerous cells; liver cell dysplasia (large dysplastic cells), as described by Anthony et al., should not be considered as a true preneoplastic change. PMID- 3038719 TI - [Present state and prospectives in the diagnosis and therapy of lung neoplasms. 6. Internal therapy--with special reference to chemotherapy of small cell carcinoma]. PMID- 3038720 TI - [Desipramine-induced down regulation of beta-adrenergic receptors: effects of noradrenergic and serotonergic neuronal activities and of alpha 2-adrenergic receptor mediated mechanisms]. AB - The mechanisms of antidepressant (desipramine)-induced down regulation of beta adrenergic receptors were investigated. The number of beta-adrenergic receptors in rat cerebral cortex was increased by the destruction of norepinephrine (NE) neurons with 6-hydroxydopamine or DSP-4 pretreatments. Desipramine (DMI)-induced down regulation of beta-receptors was completely prevented by these procedures, whereas the destruction of serotonin (5-HT) neurons neither changed the receptor number nor prevented the DMI-induced down regulation. The number of beta adrenergic receptors showed a decrease within 3 days by treatment of DMI plus (+) mianserin or yohimbine which promotes NE and 5-HT release from nerve endings through presynaptic alpha 2-adrenergic antagonism. Another alpha 2-adrenergic antagonist (-)mianserin, which promotes only 5-HT release, also accelerated the down regulation induced by DMI, but this acceleration was not prevented by the destruction of 5-HT neurons. Combination of DMI with 5-HT releasing agent methiothepin (5-HT autoreceptor antagonist) did not accelerate beta-receptor down regulation. These results suggest that 1) NE availability in the synaptic cleft plays a significant role in the regulation of beta-receptor number, but 2) postsynaptic alpha 2-adrenergic mechanisms also seem to contribute to the regulation, whereas 3) 5-HT neurons have no significant effect on the process. PMID- 3038721 TI - Plasma GABA, GABA-like activity and the brain GABA-benzodiazepine receptor complex in rats with chronic hepatic encephalopathy. AB - Plasma gamma-aminobutyric acid (GABA)-like activity, plasma GABA and the brain GABA-benzodiazepine receptor complex were studied in rats with chronic hepatic encephalopathy. Portal vein ligation (after prior subcutaneous transposition of the spleen) results in complete portal bypass of splanchnic blood. In addition, significant hepatocellular damage was superimposed on this model of portosystemic shunting by ligation of the common bile duct. Plasma GABA-like activity (determined by radioreceptor assay) and true plasma GABA concentrations (determined by high-performance liquid chromatography) were found to be significantly increased in both portal vein-ligated and portal vein and bile duct ligated rats, compared with controls. However, plasma GABA-like activity was consistently greater than the concentration of true GABA in the plasma of all rats. This suggested the presence of a GABA-like factor in plasma that can inhibit [3H]GABA binding, but is not GABA itself. The concentration of this GABA like factor was significantly increased in the plasma of rats with chronic hepatic encephalopathy. Despite the significant increase in plasma GABA-like activity and plasma GABA concentrations, there was no alteration in the affinity or density of the physiologically relevant, low-affinity brain GABA binding site in the rats with portal vein ligation, with or without bile duct ligation. There was also no significant alteration in brain benzodiazepine binding in these rats. GABA enhancement of benzodiazepine binding was unchanged in the portal vein ligated rats. However, the maximal enhancement of benzodiazepine binding was decreased in the rats with portal vein and bile duct ligation. There appears to be no substantial evidence that brain GABAergic neurotransmission is increased in these models of chronic hepatic encephalopathy. PMID- 3038723 TI - Aflatoxin content of foods served to a population with a high incidence of hepatocellular carcinoma. AB - During three seasons, 36 samples of foods served to mentally retarded clients with a high incidence of hepatocellular carcinoma were analyzed for aflatoxin. Aflatoxin was not detected (less than 5 ppb) by thin-layer liquid chromatography in 35 food samples containing peanuts, corn, wheat or milk. One peanut butter sample contained 20 ppb aflatoxin. Aflatoxin content of these foods was at or below the level permitted by the Food and Drug Administration. Aflatoxin is probably not responsible for liver disease in this population. PMID- 3038722 TI - Diminished responsiveness of male homosexual chronic hepatitis B virus carriers with HTLV-III antibodies to recombinant alpha-interferon. AB - In a randomized controlled trial, 41 chronic hepatitis B virus carriers were allocated, by opening numbered computerized randomization envelopes, to receive recombinant interferon-alpha 2A at three different doses: 2.5; 5.0, and 10.0 mU per m2. Thirty-two patients received treatment (6 for 3 months, 26 for 6 months), and 9 patients were controls (received no treatment). Ninety-three per cent of our patients were homosexual, and 41% had anti-HTLV-III in their serum. None of the control patients lost HBeAg. In contrast, six of the anti-HTLV-III-negative patients (33%) responded to treatment (p less than 0.02): five of these responders were homosexual (p less than 0.05). The response rate was greatest (44%) in the anti-HTLV-III-negative patients who received 10 mU per m2 of recombinant interferon-alpha 2A. None of the anti-HTLV-III-positive patients responded to treatment. The percentage reduction of hepatitis B virus DNA was significantly less in the anti-HTLV-III-positive group in comparison to the anti HTLV-III-negative group at 1 and 4 months of treatment and at 3 months after the end of treatment (p less than 0.05). These patients were younger (33 vs. 42 years, p less than 0.02), had lower mean baseline AST values (42 vs. 80 IU per liter, p less than 0.02) and tended to have milder histological disease. Homosexual men with HBeAg-positive chronic liver disease who are anti-HTLV-III positive appear to be less responsive to the direct antiviral and immunomodulatory effects of recombinant interferon-alpha 2A. This may be due to the subclinical immunosuppressive effects of co-infection with HTLV-III. PMID- 3038724 TI - RNA transcripts of hepatitis B virus in hepatocellular carcinoma. AB - The states of hepatitis B virus DNA, RNA transcripts and antigens (HBsAg and HBcAg) were studied in the liver with hepatocellular carcinoma by blot hybridization and immunohistological methods. We used whole hepatitis B virus DNA and gene specific probes (S, C and X genes) for hybridization. Integrated viral DNA was found in all five tumors, but episomal DNA was not detected in the neoplastic tissue. In the nonneoplastic cirrhotic liver, episomal DNA was found in four cases and integrated viral DNA in four cases. A large amount of hepatitis B virus-specific RNA transcripts was demonstrated in nonneoplastic liver of all five cases. Two major transcripts, 2.4 and 3.4 kb in length, were identified. The former hybridized with the S gene probe and the latter with the C gene probe, suggesting that they were messenger RNAs for HBsAg and HBcAg, respectively. In contrast to nonneoplastic liver, RNA transcripts were found in the neoplastic portion in only two cases, small in quantity; they primarily hybridized with S and X, but not with C genes. Novel species of 4.0 and 3.9 kb transcripts were found in the neoplastic and the nonneoplastic portions, respectively, in one case. They may represent fusion transcripts of integrated viral DNA and cellular flanking sequences. PMID- 3038725 TI - Relation of the hepatitis B virus carrier state to hepatocellular carcinoma. AB - The attempt to divide the large group of chronic HBsAg carriers into "healthy" vs. those with chronic hepatitis of various intensities is sometimes difficult. The major problems are overlap in clinical manifestations, hepatic test results and histologic as well as virologic features. Nevertheless, this separation is not only conceptually important, but may also be useful in patient management, particularly because of the risk of transition to cirrhosis and HCC. Although at least 75% of patients with HCC associated with HBV have cirrhosis, the time point at which the cirrhosis developed is not established, particularly since the vast majority of chronic HBsAg carriers fall into the "healthy" category. Important unanswered questions are, therefore: how often do "healthy" carriers develop cirrhosis and/or HCC, including the time relations between the two? Does the transformation to HCC result from one or several identifiable acute events in the "healthy" carrier (or in mild CPH) or is it a gradual process of progressing chronic hepatitis B in which intercurrent exacerbations may still play a role? Do the quantitative observations as to the relation between persistent HBV infections and HCC in the East apply to Western countries? Our hypothesis concerning pathogenesis is based on pathologic, molecular, clinical and epidemiologic observations and concepts, and is supported by studies of hepadna virus-infected animals. This thesis proposes that integration of HBV DNA into host chromosomes in acute or chronic hepatitis or during the "healthy" carrier state corresponds to an initiation event similar to that described in chemical carcinogenesis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3038727 TI - Soft tissue sarcomas apparently arising in chronic tropical ulcers. AB - Of 720 soft tissue sarcomas received from Malawi over a 15 year period, eight had apparently arisen in chronic tropical ulcers which are endemic in that part of Africa. These eight tumours were classified as leiomyosarcoma (three), extraskeletal osteosarcoma (two), malignant fibrous histiocytoma (one), myxoid liposarcoma (one) and unclassifiable (one). All had a history suggestive of malignant change in a long-standing ulcer, in each case clinically thought to be a squamous carcinoma. However, in none was there evidence of an epithelial origin and all were negative for epithelial membrane antigen and cytokeratin. This association has not previously been reported. The validity of this proposed association is discussed. PMID- 3038726 TI - The role of an albumin receptor in hepatic organic anion uptake: the controversy continues. PMID- 3038728 TI - Oncocytic glomus tumour: a new variant. AB - A new variant of glomus tumour characterized by oncocytic change is reported. The light and electron microscopy and immunohistological findings are described. This is the first reported case of an oncocytoma of non-epithelial origin. PMID- 3038729 TI - Oncocytic liver cell adenoma. AB - A 46-year-old woman underwent right hepatic lobectomy for the removal of a solitary liver tumour from a non-cirrhotic liver. The macroscopic and light microscopic features were those of a liver cell adenoma but the cells appeared oncocytic. Electron microscopy confirmed the numerous mitochondria characteristic of oncocytes. The difficulty of distinguishing oncocytic liver cell adenoma from fibrolamellar hepatocellular carcinoma is discussed. PMID- 3038730 TI - Ten families with fragile X syndrome: linkage relationships with four DNA probes from distal Xq. AB - We present clinical, cytogenetic, and linkage data of four DNA probes from the terminal long arm of the X chromosome in ten new families with fragile X syndrome. A prior/posterior method of multipoint linkage analysis is employed to combine these results with published data to refine the linkage map of terminal Xq. Ten possible probe/disease orderings were tested. The order with the greatest posterior probability (0.78) of the five loci is 52a-F9-fragile X gene-DX13-St14, although the order with reversal of the positions of 52a and F9 has a posterior probability 0.15. The mean estimates of the distances between the probes and the fragile X gene are 38 cM and 33 cM for the proximal probes 52a and F9, and 8 cM and 12 cM for the distal probes DX13 and St14. Although the current method of choice in the prenatal diagnosis and carrier detection of the fragile X syndrome remains detailed cytogenetic analysis, consideration is given to the potential role of these DNA probes, both singly and in pairs. PMID- 3038731 TI - Alpha-thalassemia in Saudi Arabia: deletion pattern. AB - Alpha-Thalassemia exists at a high prevalence in several regions of Saudi Arabia. The restriction endonucleases BamHI and BglII were used to investigate the molecular basis of deletion type of alpha-thalassemia in 226 subjects from the eastern and 61 subjects from the northwestern regions of the country. The arrangements -alpha/alpha alpha and -alpha/-alpha were common. BglII digestion revealed the existence of rightward deletion in a majority of the cases. Leftward deletions, both homozygous and heterozygous, were also identified. Triple alpha gene arrangements alpha alpha alpha anti 3.7/ were observed at a low frequency in both regions. PMID- 3038733 TI - [Sulbactam and clavulanic acid: studies of enzyme kinetics and synergism with ampicillin and mezlocillin]. AB - Both inhibitors clavulanic acid and sulbactam exhibit high affinity (Ki-values less than 10(-6) mol/l) to the beta-lactamases of gram-negative bacteria with predominant penicillinase activity and lowered affinity to enzymes with cephalosporinase activity (above all clavulanic acid). As compared to sulbactam there was a stronger synergism of clavulanic acid with ampicillin or mezlocillin in those isolates producing either a plasmid-mediated or a chromosomally-mediated penicillinase. In beta-lactamase-overproducing Klebsiella ssp. variants both inhibitors could protect neither ampicillin nor mezlocillin from breakdown independent of the synergy observed in the corresponding wild-type strains. Moreover, there was a marked synergism between mezlocillin (but not ampicillin) and sulbactam in beta-lactamase-overproducing Enterobacter cloacae variants. There was no strong relation between the amount of enzyme produced and the resulting synergy between inhibitor and antibiotic among clinical isolates. These observations agree with the results obtained for routinely performed susceptibility testing of amoxycillin/clavulanic acid and ampicillin/sulbactam. PMID- 3038732 TI - Absence of Y-specific DNA sequences in human 46,XX true hermaphrodites and in 45,X mixed gonadal dysgenesis. AB - A search for Y-specific DNA sequences has been performed in a sample of seven 46,XX true hermaphrodites and one 45,X mixed gonadal dysgenesis case and compared with a sample of 11 XX males. Using six Y-specific DNA probes no hybridization signal was obtained in the hermaphrodite group; in contrast, all XX males gave a positive signal with at least one probe. This difference is statistically highly significant. We conclude that the aetiology of true hermaphroditism is different from that of the XX male syndrome. As all cases of the hermaphrodite group are positive for the serological sex-specific antigen (Sxs) it is concluded that this antigen can be present even in the absence of Y-specific DNA. PMID- 3038734 TI - An enzyme immunoassay to detect Australian flaviviruses and identify the encephalitic subgroup using monoclonal antibodies. AB - An antigen-capture enzyme-linked immunosorbent assay (ELISA) has been developed to detect antigens of Australian flaviviruses in mosquito pools, suckling mouse brain and infected cell culture supernatant fluid. A monoclonal antibody reactive to an epitope on the envelope glycoprotein common to all flaviviruses was used as the capture antibody. Purified rabbit IgG, produced against Murray Valley encephalitis (MVE) virus, which reacted with eight Australian flaviviruses in haemagglutination inhibition (HI) and in an indirect fluorescent antibody test, was used as the indicator antibody in direct and indirect antigen-capture ELISA. A monoclonal antibody specific for a subgroup of encephalitic flaviviruses was conjugated to horseradish peroxidase and used as the indicator antibody to distinguish MVE, Kunjin and Alfuy viruses from the remainder tested. This ELISA could detect viral antigen in mosquito cell culture fluids and suckling mouse brain preparations at titres as low as 1000 TCID50/100 microliters. Viral antigen in a single mosquito infected with MVE could be detected in a pool of 500. PMID- 3038735 TI - DNA sequences of human papillomavirus types 11, 16 or 18 in invasive cervical carcinoma of Western Australian women. AB - The presence of human papillomavirus (HPV) types 11, 16 and 18 in 77 biopsies of cervical intraepithelial neoplasia (dysplasia) and carcinoma of the uterine cervix of a sample of women from Western Australia was examined using "Southern" blot hybridisation. HPV-DNA was found in 17 of the 23 dysplasias and 43 of the 54 invasive carcinomas examined but not in the 5 biopsies obtained from areas assessed as normal by colposcopy and histology. Five of 11 biopsies of mild to moderate dysplasias contained HPV type 11 (HPV-11), 2 HPV-16 and 1 HPV-18. Of 12 severe dysplasias/carcinoma in situ, 2 contained HPV-11, 4 HPV-16 and 2 HPV-18. One biopsy contained both HPV-11 and HPV-16. Of 45 squamous cell carcinomas examined for HPV-DNA, 24 contained HPV-16, 5 HPV-11 and 1 HPV-18. Both HPV-11 and HPV-16 were found in 6 of the squamous cell carcinomas and 2 contained both HPV 16 and HPV-18. Of 6 adenosquamous carcinomas examined, 3 contained HPV-DNA, 2 with HPV-16 and 1 with HPV-11. HPV types 16 or 18 were also found in 2 of 3 adenocarcinomas. This study shows a strong association between the papillomavirus and uterine cervical cancer in a sample of women from Western Australia. HPV-16 was more frequently associated with severe dysplasia and cancer than with mild or moderate dysplasia supporting the view that this HPV genotype may have a greater oncogenic potential than HPV-11. PMID- 3038736 TI - Human T lymphocyte colonies require IL2 and are inhibited by anti-Tac. AB - Human T lymphocyte colony formation can be induced in a two-step culture system with phytohaemagglutinin (PHA) as the only exogenous growth factor. In this system the first step is the overnight pre-incubation of mononuclear cells in a liquid medium in the presence of PHA ('activation step'). The second step is the seeding of these PHA-activated cells into semi-solid agar, again in the presence of PHA, and their culture for 7 days ('proliferative step'). We have previously proposed that interleukin 2 (IL2) is produced during the second (proliferative) step and that this IL2 facilitates the production of colonies from PHA-activated colony-forming cells. Here we substantiate this proposal by demonstrating that both highly purified IL2 and recombinant IL2 stimulate T colony formation from PHA-treated cells. The monoclonal antibody anti-Tac, which is specifically directed against the IL2 receptor, dramatically inhibited colony formation when it was included during the agar culture (proliferative step). There was no effect when anti-Tac was included during the liquid pre-incubation culture (activation step). These results provide strong evidence that IL2 is the critical factor in the development of T lymphocyte colonies at the stage of conversion of PHA activated cells into colonies. PMID- 3038737 TI - The cytotoxic response to murine cytomegalovirus: requirements for the generation of MCMV-specific target cells. AB - The preparation of target cells susceptible to lysis by murine cytomegalovirus (MCMV)-immune T cells in vitro was investigated and found to be dependent upon target cell type, culture conditions, virus adsorption protocol and virus preparation. Optimally sensitive MCMV-infected targets were obtained by preculture of mouse embryo fibroblasts (MEF) in 3% vol/vol concanavalin-A stimulated spleen cell supernatant (CSS)-supplemented medium, adsorption of salivary gland stock MCMV under 800 g centrifugation and at least 4 h further incubation at 37 degrees before addition of T cells. In contrast, salivary gland stock MCMV did not cause thioglycollate-induced peritoneal exudate cells to be susceptible to MCMV-specific T cell-mediated lysis, whereas tissue culture passaged stock MCMV was successful. The preparation of MCMV-infected target cells is discussed in terms of the need for H-2 and viral antigen expression for T cell recognition. PMID- 3038738 TI - Herpes zoster: multicentric. PMID- 3038739 TI - Alteration of immune response to coxsackie B3 virus by streptozotocin in dual aetiology diabetes mellitus in mouse. PMID- 3038741 TI - Hepatocellular carcinoma--a study of 200 cases. PMID- 3038742 TI - Role of resident peritoneal macrophages in age-related susceptibility to Japanese encephalitis virus in mice. PMID- 3038740 TI - Inheritance of a mutant histocompatibility gene and a new mammary tumor virus genome in the B6.KH-84 mouse strain. AB - The potential association between integration or deletion of mouse mammary tumor virus (MMTV) retroviral sequences and the appearance of non-H-2 histocompatibility (H) antigen mutations was investigated. Genomic blots from inbred strains carrying 22 loss, gain-loss, and gain mutations on the BALB/c and C57BL/6 backgrounds were hybridized with probes homologous to the long terminal repeat (LTR) and envelope (env) regions of MMTV. Twenty-one mutants were identical in restriction patterns to the respective background strains with all tested restriction enzymes and both probes. However, genomic blots of one gain mutant, B6.C-KH-84, exhibited restriction fragments which were not exhibited by either of the parental strains, C57BL/6 or BALB/c. An additional 5.5 kb Eco RI fragment was observed with the env probe and additional 9.2 kb and 5.5 kb fragments were observed with the LTR probe. These observations were substantiated by hybridization of these two probes with genomic blots generated with additional restriction enzymes. Assuming that the new provirus contains a single, internal Eco RI site as has been observed for other MMTV proviral sequences, it is presumed that the new provirus includes both 5' and 3' LTRs in addition to the env region. Based on the unique sizes of the observed restriction fragments relative to other identified MMTV proviral sequences, this provirus has been designated Mtv-22. The potential role of Mtv-22 in the genesis of the gained histocompatibility antigen in B6.C-KH-84 is discussed. PMID- 3038743 TI - Role of HSV antibodies in precancerous & cancerous lesions of the uterine cervix- a prospective study. PMID- 3038744 TI - Comparison of sodium citrate with magnesium trisilicate as pre-anaesthetic antacid in emergency caesarean sections. PMID- 3038745 TI - Decreased intraocular pressure in dogs with one-kidney, one wrapped hypertension. AB - We examined the relationship of intraocular pressure and the development of one kidney, one wrapped (perinephritic) hypertension in the dog. Conscious femoral arterial pressure (direct arterial puncture) and intraocular pressure (Schiotz tonometer) were measured weekly before and after the surgical induction of hypertension in 11 healthy male mongrel dogs and before and after unilateral nephrectomy in 15 normotensive control dogs. Preoperative mean arterial pressure (102 +/- 5 vs 99 +/- 8 [SD] mm Hg, hypertensive vs control dogs) and intraocular pressure (18.1 +/- 2.5 vs 17.7 +/- 2.1 mm Hg, hypertensive vs control dogs) were similar in both groups. In normotensive control dogs, mean arterial pressure and intraocular pressure averaged over the postoperative period (4-8 weeks) did not differ significantly from preoperative values. In contrast, during the same period arterial pressure significantly increased and intraocular pressure significantly decreased in hypertensive dogs (arterial pressure, 163 +/- 8 mm Hg; intraocular pressure, 11.9 +/- 4.0 mm Hg; p less than 0.001 for both values compared with corresponding values in control dogs). Intraocular pressure was inversely related to arterial pressure in hypertensive dogs (r = 0.56, p less than 0.01). These observations indicate that intraocular pressure decreases with the development of canine one-kidney, one wrapped hypertension. The mechanism of this decrease may be related to abnormalities in Na+,K+-adenosine triphosphatase activity found in this form of hypertension. PMID- 3038746 TI - Effect of beta-adrenergic receptor blockade on atrial natriuretic peptide in essential hypertension. AB - Plasma levels of atrial natriuretic peptide (ANP) were measured in 32 untreated subjects with essential hypertension and in 31 patients undergoing long-term treatment with beta-blockers. Patients receiving beta-blockers had significantly higher mean plasma ANP levels (72.0 +/- 36.0 [SD] pg/ml) than did untreated hypertensive subjects (39.8 +/- 15.8 pg/ml; p less than 0.01) and healthy normotensive controls (33.9 +/- 16.6 pg/ml; n = 61, p less than 0.01), while the mean plasma ANP concentration in untreated hypertensive subjects was not statistically different from that in control subjects. Administration of atenolol, 50 mg/day, for 4 weeks to 10 untreated subjects resulted in a significant (p less than 0.001) rise in plasma ANP levels (from 38.8 +/- 9.5 to 68.7 +/- 20.6 pg/ml). In 31 patients undergoing long-term treatment with beta blockers, multivariate regression analysis revealed that age, pretreatment mean blood pressure, and plasma concentration of cyclic 3',5'-guanosine monophosphate (cGMP) were significant predictors of plasma ANP levels. These results suggest that beta-adrenergic receptor blockade in patients with essential hypertension elevates plasma ANP levels with a concomitant rise in cGMP concentrations, and that increased ANP in plasma may play a role in the compensatory mechanism that operates in response to beta-adrenergic receptor blockade. PMID- 3038747 TI - Epidemic poliomyelitis. PMID- 3038748 TI - [Communicable diseases: various manifestations of herpetic disease]. PMID- 3038749 TI - [The activity of a chlorhexidine mouthwash in the production of free radicals. Demonstration of anti-inflammatory activity]. PMID- 3038750 TI - Effects of a phagocytosis-stimulating factor derived from polymorphonuclear neutrophils on the functions of macrophages. AB - The effects of phagocytosis-stimulating factor (PSF) derived from polymorphonuclear neutrophils on macrophage functions were studied. PSF enhanced the initial rate of phagocytosis of serum-opsonized zymosan particles by macrophages, whereas it did not affect the phagocytosis of immunoglobulin G sensitized and inert zymosan particles. Kinetic studies showed that PSF accelerated the ingestion step, but not the attachment step, of phagocytosis by macrophages. On the other hand, PSF did not affect the other macrophage functions such as O2- generation, chemotaxis, adherence, and enzyme release. These results suggest that PSF may specifically modulate the complement receptor function of macrophages. Immunoblot assay showed the absence of components in macrophage which reacted with purified antibodies against polymorphonuclear neutrophil derived PSF, and an extract from phagocytosing macrophages had no phagocytosis stimulating activity, indicating that the macrophages did not produce PSF-like substances. PMID- 3038751 TI - Influence of type and opsonization of ingested particle on intracellular free calcium distribution and superoxide production by human neutrophils. AB - Intracellular free calcium concentration [( Ca2+]i) was measured with the fluorescent Ca2+ indicator Fura 2 within individual human neutrophils during the phagocytosis of several types of particles, including serum-treated zymosan (STZ), immunoglobulin G (IgG)-coated zymosan (IGZ), C3b-coated zymosan (C3Z), nontreated zymosan (Z), and serum-treated similarly sized latex particles (STL). STZ was coated with both IgG and C3b. IGZ was coated with only IgG, and C3Z was coated with only C3b. STL was coated with only C3b but to a lesser extent than C3Z. The ingestion of particles was greatest for STZ and somewhat lower for C3Z. Ingestion of IGZ and STL was much less than ingestion of C3Z. The relative efficiencies of the particles for inducing superoxide production were as follows: STZ greater than IGZ = C3Z greater than Z = STL. [Ca2+]i significantly increased from the resting level of approximately 70 to greater than 240 nM (P less than 0.01) during phagocytosis of the particles. The increment in [Ca2+]i was greater in the paraphagosomal region than in the cell body after the ingestion of STZ or IGZ. The mean peak [Ca2+]i values in the paraphagosomal cytoplasm of neutrophils ingesting one particle of STZ, IGZ, C3Z, Z and STL were 536.1 +/- 57.6, 424.7 +/- 55.8, 373.8 +/- 62.7, 272.3 +/- 31.5, and 270.8 +/- 38.0 nM, respectively, which showed good correlation (r = 0.97) with the efficiency of the particles for inducing superoxide production. Depletion of extracellular Ca2+ by EGTA [ethylene glycol-bis(beta-aminoethyl ether)-N,N,N'N'-tetraacetic acid] attenuated both [Ca2+]i increase and superoxide production induced by particles. Thus [Ca2+]i increased after ingestion of several types of particles, and the subcellular pattern of [Ca2+]i was different depending on the type of particle ingested. Greater increases in paraphagosomal [Ca2+]i were closely associated with greater increases in superoxide production by neutrophils. PMID- 3038752 TI - Hypochlorite scavenging by Pseudomonas aeruginosa alginate. AB - Alginic acid was purified from a mucoid clinical isolate of Pseudomonas aeruginosa. Luminol-dependent chemiluminescence of phorbol myristate acetate stimulated neutrophils was inhibited by this alginate, but oxygen consumption was unaffected. Further studies indicated that this effect was due to the ability of the pseudomonal alginate to scavenge hypochlorite. A seaweed alginate was less effective and dextran T500 was ineffective in hypochlorite scavenging. It appears that the uronic acid core and the O-acetyl groups of pseudomonal alginate are involved in its hypochlorite-scavenging ability. The relevance of this phenomenon was demonstrated by the greater resistance to killing by hypochlorite of mucoid P. aeruginosa compared with a nonmucoid revertant, and the addition of purified alginate to the nonmucoid revertant protected the organism from hypochlorite. Thus, this extracellular polysaccharide may enhance the virulence of P. aeruginosa by scavenging the phagocyte-generated oxidant HOCl. This enhanced virulence may be involved in disease processes in which mucoid organisms predominate, such as cystic fibrosis. PMID- 3038754 TI - Eosinophil- and neutrophil-mediated injury of human lung fibroblast cells. AB - The effect of culturing purified rat peritoneal neutrophils and eosinophils with the human foetal lung fibroblast cell line, MRC-5, was studied. Target cell damage was measured by the failure of the MRC-5 cells to form confluent monolayers during a 6-day incubation period. Neutrophils were more effective at inhibiting cell growth than were eosinophils with greater than 50% inhibition being recorded at initial effector: target cell ratios of 5:1 using neutrophils and 40:1 using eosinophils. Following stimulation with phorbol myristate acetate (PMA), one quarter the number of eosinophils was required to give 50% inhibition of cell growth. The addition of either catalase or superoxide dismutase partly reduced the activity of PMA-stimulated eosinophils, although only with catalase did this reach significance. PMA-stimulated neutrophils did not show any enhanced activity against the MRC-5 cells. Addition of the bacterial analogue f-Met-Leu Phe (10(-7)M) was unable to increase the cytotoxic activity of eosinophils. Disrupted eosinophil suspensions were as effective as intact cells at inhibiting the growth of target cells whereas some loss in effectiveness was seen with disrupted neutrophils. Membrane-free supernatants from both eosinophils and neutrophils were inactive. The results suggest that although eosinophils and neutrophils are both capable of damaging lung fibroblast cells in culture, the latter is more effective, eosinophils appearing to require an additional stimulation in vitro. Possible mechanisms of cytotoxicity are discussed. PMID- 3038753 TI - Identification of a specific receptor for plasmin on a group A streptococcus. AB - Certain group A streptococci demonstrate surface receptors that bind selectively to the key fibrinolytic enzyme, plasmin. These bacteria show no reactivity with the zymogen protein plasminogen or with other serine class proteases, such as trypsin or urokinase. Bacterium-bound plasmin retains its ability to cleave synthetic substrates and its ability to hydrolyze a fibrin clot. The bacterium bound plasmin is not effectively regulated by its physiological regulator, alpha 2-plasmin inhibitor. This study is the first report of a bacterium-associated receptor for plasmin. PMID- 3038755 TI - Defective cell-mediated response to EBV-transformed B cells in a healthy individual with regular EBV antibody titers. AB - We have analyzed the EBV-related immune parameters of a healthy EBV-seropositive individual (ST) who has regular antibody titers but defective inhibitory capacity toward the growth of autologous EBV-infected B cells. This in vitro function reflects the EBV-specific memory because it does not occur in experiments performed with cells of seronegative individuals. An analysis of events following in vitro EBV infection showed that lymphocytes of ST behaved in some tests in the same way as those collected from seronegative individuals. These parameters were: lack of gamma-IFN production 24 hr after EBV infection; low production of soluble factors that inhibit EBV-induced B-cell proliferation; lack of generation of LCL selective cytotoxicity after repeated stimulation with autologous LCL; and high proportion of EBNA-positive cells in 7-day-old EBV-infected cultures. On the other hand, cellular memory to the virus detected by the production of IL-2 24 hr after infection, and by the production of LIF upon exposure to EBV-encoded antigens, conformed with the results obtained with seropositive individuals. T cell-mediated inhibition of EBV-induced B-cell growth in vitro has been regarded as a corollary of in vivo control of EBV-infected B cells. However, it is absent or has a low efficiency in certain disease categories which are not accompanied by risk of B-EBV growth. Our results with a healthy individual also indicate that several mechanisms contribute to a harmless life-long virus carrier state. PMID- 3038756 TI - Colorectal polyps and diet: a case-control study in Marseilles. AB - This study investigates the differences in usual past diet between 252 subjects with newly diagnosed adenomatous or villous polyps of the colon and rectum and a group of 238 hospital controls. Cases and controls were interviewed in hospital by 3 nutritionists using a dietary history questionnaire focused on the diet during the preceding year. Nutrient intake was estimated by means of ad hoc food tables adapted from French and British tables. Out of 16 food groups considered in the analyses, the cases reported lower consumption of oil and potatoes and higher consumption of sugar added to food and drink. Among nutrients, we found that cases had a lower consumption of carbohydrates (not taking into account added sugar), potassium, magnesium and vitamin B6. We found a slightly lower intake of fibre and a slightly higher intake of saturated fat among cases, though neither was statistically significant. Intake values for fibre and for carbohydrates were highly intercorrelated and, due to measurement errors, the effect of one may be masked by the other and vice versa. The hypothesis that some components of carbohydrates (starches, fibre and natural sugars but not added sugar) play a protective role in relation to the biology of tumours of the intestinal tract is considered in further multivariate analyses and in the "Discussion". PMID- 3038757 TI - Increased prevalence of human papillomaviruses in the lower genital tract of pregnant women. AB - In order to evaluate the influence of pregnancy on the presence of human papillomavirus (HPV) in the lower female genital tract, cervical smears of 92 pregnant and 96 non-pregnant women, matched by age, were examined for the presence of HPV-DNA by means of Southern blot hybridization. All patients had negative PAP smears. Twenty-six (28%) of the pregnant women and 12 (12.5%) of the non-pregnant women were positive for HPV. HPV 16 accounted for 42% of all positive pregnant cases and only 25% of the positive non-pregnant cases. Smears of pregnant patients contained more than 10 pg viral DNA in 45% of the cases against 20% in the non-pregnant group. HPV 16 showed the most active replication in both groups. This study demonstrates an increased prevalence of HPV (preferentially of HPV 16) and a higher replication rate of viral DNA during pregnancy. PMID- 3038758 TI - Conversion of the lymphoma line "BJAB" by Epstein-Barr virus into phenotypically altered sublines is accompanied by increased c-myc mRNA levels. AB - The steady-state level of c-myc proto-oncogene mRNA was investigated in the EBV negative human B-lymphoma line BJAB and 2 sublines that have been converted by EBV into stable EBV-genome-carrying and EBNA-positive status. The EBV-converted sublines expressed c-myc at a 2- to 6-fold higher level than the original BJAB during exponential growth. The EBV-positive BJAB lines are known to differ from the parent line in several phenotypic characteristics, including increased agarose clonability, lower serum requirement and, in one case, increased tumorigenicity in nude mice. The pattern of increased c-myc expression accompanying EBV conversion was not observed in the myc/Ig translocation-carrying Burkitt lymphoma line RAMOS or in 2 of its EBV-converted sublines. PMID- 3038759 TI - Loss of basement membrane deposits and development of invasive potential by virally-transformed rat mammary cells are independent of collagenase production. AB - The myoepithelial-type cell line, Rama 712, derived from a normal rat mammary gland, deposits an extracellular matrix containing type-IV collagen and other basement membrane proteins round its cellular periphery. After transformation with a temperature-sensitive mutant of Rous sarcoma virus (tsRSV) the cells fail to deposit an extracellular matrix at the permissive temperature (35 degrees C), but retain the capacity to do so at the non-permissive temperature (41 degrees C). The synthesis of type-IV collagen is not affected by the temperature shift. Rama 712 cells fail to form tumours in syngeneic rats. However, Rama 712-tsRSV cells form tumours that are locally invasive but fail to metastasize. In histological sections, the tumour cells stain with an antibody to type-IV collagen, but do not deposit any extracellular type-IV collagen. Cells isolated from the tumours (Rama 712T) remain temperature-sensitive for the extracellular deposition of type-IV collagen when grown in vitro. Rama 712, Rama 712-tsRSV and Rama 712T fail to produce any detectable type-I or type-IV collagenase at either 35 degrees C or 41 degrees C. These results show that in this system extracellular deposits of basement membrane proteins are lost from invasive tumours produced by myoepithelial-type cells by mechanisms other than those due to the production of collagenolytic enzymes. PMID- 3038760 TI - Peripheral benzodiazepines enhance the respiratory burst of macrophage-like P388D1 cells stimulated by arachidonic acid. AB - P388D1, a murine cell with macrophage properties, responds to exogenous arachidonic acid with superoxide anion production. This oxidative burst is enhanced by peripheral and mixed type benzodiazepines and this stimulation is specifically reversed by the peripheral antagonist PK 11195. In contrast, PK 11195 is unable to antagonize a stimulation caused by a non-benzodiazepine ligand such as the chemotactic peptide fMet-Leu-Phe. The optimal concentrations were close to 10 nM and corresponded to the affinities of the compounds for the peripheral benzodiazepine receptor detected on these cells. Compared to other tissues where peripheral benzodiazepines acted only at micromolar concentrations, the macrophage with its functional receptor appears as a privileged site of action for these molecules. PMID- 3038761 TI - Influence of lysophospholipids and PAF on the oxidative burst of PMNL. AB - Lysophosphatidylcholine (LC), platelet activating factor (PAF) and its precursor lysophosphatidalcholine (LP) enhance O-2-release by polymorphonuclear leucocytes (PMNL) triggered by PMA whereas lysophospholipids with other polar headgroups fail to do so. The generation of these lysophosphatidylcholine-like molecules appears to represent an essential step in the activation of the oxidative burst of the PMNL triggered by PMA since inhibition of phospholipase A2 (PLA2) by p bromophenacylbromide (BB) or mepacrine results in an inhibition of the O-2 release. This inhibition seems to be due to the reduced generation of the phospholipids studied as it could be reversed by LP. In addition, stimulation of the oxidative burst of the PMNL by the chemotactic stimuli, N-formyl-methionyl leucylphenylalanine (FMLP), and the complement fragment C5a could also be significantly enhanced by LP as shown by chemiluminescence. However, the response to the phagocytic stimulus, opsonized zymosan (Zx), is not affected by LP. These data provide evidence for the participation of phospholipid metabolism in the initiation of the oxidative burst of PMNL induced by the soluble monomeric stimuli PMA, FMLP and C5a. PMID- 3038762 TI - Differential effects of marijuana components on proliferation of spleen, lymph node and thymus cells in vitro. AB - The effects of delta 9 THC and 11-OH THC on the proliferative response of murine spleen cells stimulated in vitro with the T cell mitogens Con A or PHA were compared with the effects of these drugs on the mitogen-induced proliferation of murine thymus and lymph node cells. Thymus cells were found to be suppressed at lower cannabinoid concentration than either spleen or lymph node cells. However, splenic cells were more easily suppressed than were the lymph node cells. Lymphoid cell numbers were varied from 1 X 10(6) to 8 X 10(6) cells and treated with a constant dose of either THC or 11-OH THC. When suppression was noted with spleen and lymph node cells, the smallest number of cells in the assay resulted in the greatest level of suppression of cell proliferation. No significant suppression to PHA induced proliferation was found for lymph node cells at any cell number tested. Thymus cells were always more readily suppressed than spleen or lymph node cells regardless of the number of cells in culture. Furthermore, 11 OH THC suppressed the responsiveness of the thymus cells to PHA more than to Con A under the experimental conditions used. Thus, the ability of cannabinoids to induce suppression of the proliferative response of lymphoid cells to mitogens depends on the organ source of the cells, nature of the cannabinoid (THC or 11-OH THC), dose of the cannabinoid, mitogen used (PHA or Con A), and number of cells in culture.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3038763 TI - The penile condyloma cancer connection. PMID- 3038764 TI - Predictors of clinical AIDS in young homosexual men in a high-risk area. AB - One hundred and sixty-seven homosexual men in Los Angeles characterized by HIV antibody, T-cell numbers, titres to cytomegalovirus (CMV), and specific sexual practices were followed for two years for immune changes and for more than three years for development of clinical AIDS. Thirty-five per cent had antibody to HIV at baseline. The mean level of T-helper (Th) cells was significantly lower and of T-suppressor (Ts) cells significantly higher in HIV seropositives than in seronegatives. The annualized incidence of HIV seroconversion was 7%. Eight men developed AIDS, an attack rate of 14% in those with HIV antibody at baseline. A number of observations were made: T-cell alterations, except a transient elevation in Ts cells, were unusual in the absence of HIV antibody; a seropositive man with a T-cell alteration was significantly less likely to revert to 'within normal limits' than was a seronegative man; a steady decline in the number of Th cells preceded onset of clinical AIDS; the number of Ts cells remained higher in men subsequently developing AIDS than in other seropositive men; clinical AIDS occurred only in men with HIV antibody whose CMV antibody levels were above the median for the group (1:1600); and the attack rate for clinical AIDS was 50% in men with HIV antibody and elevated CMV who at baseline had either: fewer than 325 Th cells/cc, or whose Th/Ts ratio was below 0.8 (but whose levels of Th and Ts cells were within normal limits).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3038765 TI - Insulin releasing effects of adrenocorticotropin (ACTH 1-39) and ACTH fragments (1-24 and 18-39) in lean and genetically obese hyperglycaemic (ob/ob) mice. AB - An excessive insulin releasing effect of adrenocorticotropic hormone (ACTH) and ACTH fragments has been considered as a possible factor contributing to the hyperinsulinaemia of genetically obese hyperglycaemic (ob/ob) mice. To investigate this possibility, plasma glucose and insulin responses of 11- to 14 week-old fed lean and ob/ob mice were examined after intraperitoneal administration of ACTH 1-39, ACTH 1-24 and ACTH 18-39, each at a dose of 25 nmol/mouse (50-115 micrograms/mouse). ACTH 1-39 produced a marked and rapid increase of plasma insulin in both lean and ob/ob mice, the effect being much greater in the ob/ob mutant (maximum increases of 5.5 +/- 1.5 and 46.1 +/- 4.1 ng/ml at 10 min in lean and ob/ob mice respectively, P less than 0.001). In lean mice plasma glucose concentrations showed a protracted decreased (maximum decrease of 3.7 +/- 0.5 mmol/l at 120 min), whereas glucose concentrations were increased (maximum increase of 4.2 +/- 1.3 mmol/l at 60 min) in ob/ob mice. ACTH 1-24 produced qualitatively similar but generally smaller effects than ACTH 1-39, while ACTH 18-39 did not significantly affect glucose and insulin concentrations. In 24 h fasted mice, ACTH 1-39 produced similar but generally smaller effects than in fed mice. The results suggest that the effects of ACTH on glucose and insulin homoeostasis are conferred by the N-terminal 1-24 sequence, and ACTH may exert acute effects which contribute to the hyperinsulinaemia and hyperglycaemia of ob/ob mice. PMID- 3038766 TI - Kinetics of hydroxyl radical induced poly(U) strand break formation in pulse irradiated aqueous solutions in the presence of oxygen. PMID- 3038767 TI - Surgical treatment of small hepatocellular carcinomas in cirrhosis. AB - Over the last five years a policy of systematic screening for small hepatocellular carcinomas (HCC) in patients at risk has led to an increasing number of resections in patients with cirrhosis. Remarkable progress in the surgery of HCC in cirrhosis has been accomplished through: (a) a better understanding of the surgical anatomy of the liver, (b) the definition of new types of liver resection aimed at reducing the amount of parenchyma removed while still being oncologically satisfactory, (c) the reduction of intraoperative blood loss by various techniques of clamping afferent and efferent vessels, (d) the systematic use of intraoperative ultrasonography, and (e) the prevention of postoperative variceal bleeding and the formation of ascites. Results of resection of small HCC in cirrhosis have been quite impressive in Japanese series, with a low operative mortality and above 50% three-year survivals. Results in the West have been somewhat less good. Differences in the pathology of these tumours and particularly in the rate of encapsulation could account for these differences. Clearly, surgical resection has become an established treatment for small HCC in cirrhosis. More information is needed on the results of surgery in operated patients and this should be compared with the natural history of small HCC in cirrhosis in order to better define the patients who will most benefit from these operations and which tests performed at which intervals, are most reliable in screening patients at risk. PMID- 3038769 TI - Repeated hepatic resection. AB - Sixteen patients underwent repeated hepatic resections over a 16 year period. The cases were divided into two groups: group A, non-planned repeated resections (14 cases), and group B, planned repeated resections (two cases). Group A is composed of patients requiring re-resection as a result of the hepatic re-recurrence of the neoplasia (three hepatocellular carcinomas, nine metastases from colorectal carcinoma, and two metastases from carcinoid tumor). Group B is composed of two cases (hepatocellular carcinoma and metastases of leiomyosarcoma) where the extent of the disease was incompatible with radical resection in a single time thus making necessary to plan for repeated operations. The need for correct preoperative assessment of hepatic performance using CT, US and Tc 99m HIDA scan, as well as intraoperative ultrasonography is stressed. PMID- 3038768 TI - Hepatic resection for tumours in cirrhotic livers. AB - A liver resection was performed in 25 out of 36 cirrhotic patients operated on for liver cell carcinomas. In the remaining 11 cases hepatectomy was not performed mainly because of the presence of other intrahepatic neoplastic nodules or thrombi in the portal branches revealed by intraoperative echography. The operative mortality in the 25 patients operated on was 16%; the actuarial survival at three years is 58%. Liver resection was carried out using a transparenchymal procedure; in 18 cases clamping of the hepatic pedicle was performed for an average period of 15 min. Twenty patients with small tumours had a segmentary or sub-segmentary resection; intraoperative echography proved indispensable in this situation, making it possible to recognize the lesion and outline the limits of the resection. The presence of a peritumoral capsule seems to have been an important prognostic factor. PMID- 3038770 TI - Effects of ouabain and furosemide on transepithelial electrical parameters of the isolated shark ciliary epithelium. AB - Sections of the ciliary epithelium of adult sharks (Squalus acanthias) were mounted in Ussing-type chambers (area 0.2 cm2) for measurements of transepithelial potential difference (PD), short circuit current (SCC) and calculation of transepithelial resistance (R). In 15 preparations PD was aqueous side negative (-0.51 +/- 0.12 mV; SCC 18.3 +/- 2.5 microA cm-2; R 30.7 +/- 3.1 Ohm cm2). However, in 15 other preparations incubated in identical Ringer's solution PD was aqueous side positive (0.53 +/- 0.09 mV; SCC -19.6 +/- 2.3 microA cm-2; R 27.9 +/- 2.8 Ohm cm2). 10(-5) M ouabain or 10(-4) M furosemide were applied either to the aqueous or blood side of the isolated ciliary epithelium at transepithelial negative or positive PD. When the transepithelial PD was positive on the aqueous side ouabain decreased PD and SCC within 15 to 45 min. When the spontaneous PD was negative both PD and SCC decreased when ouabain was applied to the blood side. When the drug was given to the aqueous side a biphasic response (first stimulation, then inhibition) of PD and SCC was observed. Furosemide when given to the blood side (with aqueous side PD positive) or to the aqueous side (with aqueous side PD negative) decreased PD and SCC. However, a transient stimulation of both electrical parameters was observed when furosemide was applied to either the blood side (with aqueous side PD negative) or to the aqueous side (with aqueous side PD positive). The polarity and magnitude of PD and SCC probably depend on the relative activity of sodium and chloride pumps across the cell membranes of the non-pigmented and/or pigmented cell layer. However, additional transport mechanisms cannot be excluded. PMID- 3038771 TI - Autoradiographic localization of beta-adrenergic receptors in rabbit eye. AB - Using an in vitro autoradiography, beta-adrenergic receptors were localized in the rabbit eye. Autoradiograms were generated by apposition of isotope-sensitive film to slide-mounted eye sections, labelled with [125I](-)Iodocyanopindolol. A high density of silver grains was obtained in conjunctival, corneal and ciliary process epithelium. Binding sites were also present in corneal endothelium, iris epithelium, lens epithelium, choroid and extraocular muscles. In some areas, retina was also labelled. Studies with beta-adrenergic compounds showed that the majority of beta-adrenergic receptors, detectable by autoradiography, were of the beta2 type in the rabbit eye. PMID- 3038773 TI - Adenovirus 3-7, an intermediate strain of subgenus B. AB - Adenovirus 3-7, strain Takeuchi was related to both AV3 and AV7 by neutralization, but only to AV3 by hemagglutination-inhibition. DNA restriction analysis with seven endonucleases showed about equal genetic distance from both AV3 and 7 prototypes. PMID- 3038772 TI - Studies on the morphogenesis of murine cytomegalovirus. AB - Two modes of assembly of murine cytomegalovirus (MCMV) were observed in cultured mouse embryo fibroblasts, generating two morphologically different types of viral particles: monocapsid virions and multicapsid virions. The assembly of nucleocapsids appeared to be the same for both types of morphogenesis. Three successive stages of intranuclear capsid formation could be distinguished: capsids with electron-lucent cores, coreless capsids, and capsids with dense cores. Some of the capsids were enveloped at the inner nuclear membrane to form monocapsid virions, which were first detectable in the perinuclear cisterna. Other capsids left the nucleus via nuclear pores and usually entered cytoplasmic capsid aggregates that received an envelope by budding into extended cytoplasmic vacuoles, thereby forming multicapsid virions. Since the formation of multicapsid virions is not restricted to cell culture conditions and also occurs in vivo in immunosuppressed mice, multicapsid virions may play a role in the pathogenesis of cytomegalovirus infection. PMID- 3038774 TI - Transforming potential of DNA of the human PLC/PRF/5 hepatoma cell line. AB - DNA of the human PLC/PRF/5 hepatoma cell line can induce the appearance of colonies on soft agar after transfection of BK-BK cells (BALB/c mouse kidney cells partially transformed by BK virus). Blot hybridization analyses indicate that the transformants contained the hepatitis B virus DNA sequence. This sequence disappeared during serial passage of transformants. The amplification and rearrangement of the integrated BK virus genome appears to be specifically associated with transformation. PMID- 3038775 TI - Isolation of transforming Epstein-Barr virus from plasma of HTLV-III/LAV-infected patients. AB - Fresh plasma and peripheral blood mononuclear leukocytes were obtained from 11 patients with acquired immunodeficiency syndrome (AIDS). All of these patients were seropositive for antibodies to HTLV-III/LAV and Epstein-Barr virus (EBV). Plasma and uncultured peripheral blood leukocytes from these patients were tested for the presence of infectious EBV and membrane antigen (EBV-MA). Transforming EBV was isolated from the cell-free plasma of 4 patients with AIDS exhibiting EBV MA in uncultured cells. No virus was detected in plasma from the healthy donors or from the remaining AIDS patients. No neutralizing antibody to EBV was detected in the 4 patients whose plasma contained EBV. In contrast, neutralizing antibody was present in the plasma of EBV-seropositive healthy donors and in 7 other patients with AIDS. One of the AIDS patients whose plasma contained EBV recently developed Burkitt's-type lymphoma. The production of transforming EBV could be induced either directly or indirectly by HTLV-III/LAV infection, and also could be involved in the frequently observed B cell malignancy associated with AIDS. PMID- 3038776 TI - Presence of cytomegalovirus in brown fat. Study in a rat model. AB - Cytomegalovirus infection of laboratory rats resulted in the appearance of virus and viral antigens in interscapular and periaortic brown fat. This appearance was limited to acute infection (first week post-infection) and was dependent on the route of inoculation. In our study virus could only be detected in subcutaneous brown fat in young rats (till 4 weeks of age), whereas in the deep brown fat (periaortic region) no age dependency was found. PMID- 3038778 TI - Congenital infections: a prospective study. PMID- 3038777 TI - The early enhancer-promoter of BKV and host range for transformation. AB - The early genes of the human papovavirus BKV and simian virus 40 (SV40) show a different host range for transformation. The early region of SV40 efficiently transforms human fibroblast cells, whereas the early region of BKV does not. Interchanging noncoding enhancer-promoter sequences around the origin of replication between BKV and SV40 showed that the early enhancer-promoter sequences of BKV and SV40 could substitute for each other, as far as the induction of expression of tumor antigens and morphological transformation is concerned; the efficiency of transformation was influenced by the enhancer promoter sequences, and the difference in host range for transformation between BKV and SV40 was determined by the early gene products rather than by the enhancer-promoter sequences. PMID- 3038780 TI - Occurrence of cyclic AMP and related enzymes during germination of Pinus pinea seeds. AB - The occurrence of cAMP, adenylate cyclase and cAMP phosphodiesterase has been tested in Pinus pinea seed during germination. The study has been carried out on dormant and imbibed seeds, seedlings, endospermic residues, roots and cotyledons. cAMP has been detected by the protein binding method and its occurrence has been verified by HPLC detections. cAMP phosphodiesterase shows a very high activity at acidic pH, while being completely inactive at pH 7.4. At this pH value, well detectable levels of adenylate cyclase have been observed. Therefore, the classical pathway of synthesis and breakdown of cAMP, already accepted for animal and bacterial cells, seems to be operating in Pinus pinea plant too. PMID- 3038779 TI - Structure and function of adenylate kinase isozymes in normal humans and muscular dystrophy patients. AB - Two isozymes of adenylate kinase from human Duchenne muscular dystrophy serum, one of which was an aberrant form specific to DMD patients, were separated by Blue Sepharose CL-6B affinity chromatography. The separated aberrant form possessed a molecular weight of 98,000 +/- 1,500, whereas the normal serum isozyme had a weight of 87,000 +/- 1,600, as determined by SDS-polyacrylamide gel electrophoresis, gel filtration, and sedimentation equilibrium. The sedimentation coefficients were 5.8 S and 5.6 S for the aberrant form and the normal form, respectively. Both serum isozymes are tetramers. The subunit size of the aberrant isozyme (Mr = 24,700) was very similar to that of the normal human liver isozyme, and the subunit size of the normal isozyme (Mr = 21,700) was very similar to that of the normal human muscle enzyme. The amino acid composition of the normal serum isozyme was similar to that of the muscle-type enzyme, and that of the aberrant isozyme was similar to that of the liver enzyme, with some exceptions in both cases. PMID- 3038781 TI - Right intracavitary cardiac tumors. Surgical management. AB - The surgical treatment of 6 patients affected by right cardiac tumors is reported. Three of them showed a right atrial myxoma, 1 metastases from an occult embryonal carcinoma and 1 a leiomyosarcoma of the pulmonary artery. While clinical diagnosis was not reliable, the echocardiographic examination was able to assess the site of the tumor, supplying useful information for the anatomy and surgery in 5 cases out of 6; in one case, the diagnosis was reached intraoperatively. All the patients survived the surgical treatment: while those with atrial myxoma recovered completely, the others with malignant tumors died from metastases 5 to 10 months postoperatively. The importance of early diagnosis is stressed together with the role of surgery, which permits the recovery of cases with benign tumors and a longer survival of cases with malignant tumors. PMID- 3038782 TI - Role of ileorectal anastomosis in the treatment of ulcerative colitis and familial polyposis. AB - A 25-year experience with 109 patients treated with ileorectal anastomosis over 147 patients observed for ulcerative colitis and 25 patients undergoing the same surgical procedure for familial polyposis, is reported. Excellent functional results were obtained, associated to satisfactory ones with respect to inflammatory recurrence and cancer development in the rectal stump. Of 86 patients with ulcerative colitis who were followed-up, only 7 (8.1%) required proctectomy at long-term for a severe inflammatory recurrence, and 4 (4.6%) have developed a cancer in the following 3 to 20 years. Of the 17 followed-up patients, over the 25 operated for familial polyposis, only 1 (5.8%) has developed a cancer between 3 and 20 years. It is concluded that ileorectal anastomosis with a close follow-up still plays a major role because is an easier surgical procedure as compared to ileoanal anastomosis, with excellent functional results, a shorter hospitalization and mainly a lower number of severe complications, which are presently burdening the results of ileoanal anastomosis. PMID- 3038783 TI - Kininase I, kininase II and aminopeptidase levels in patients with gastrointestinal tumors. AB - Some indices of the kallikrein kinin system, namely kininase I (KI), kininase II or angiotensin converting enzyme (KII-ACE) and phenylalanine-arginine aminopeptidase (AP) were analyzed, to detect their levels in ten selected normal subjects and in 20 selected patients with colonic (n = 8) and gastric adenocarcinoma (n = 12). While AP and KI levels did not show differences in the groups under analysis, KII values showed a significant difference (P +/- 0.01), present since the early stages of the diseases and unrelated to the normal laboratory indices. PMID- 3038784 TI - Effects of platelet-activating factor on electrophysiology of isolated retinas and their inhibition by BN 52021, a specific PAF-acether receptor antagonist. AB - The effects of (R)PAF-acether have been tested on the isolated rat retina model. Results indicate that (R)PAF-acether inhibits the electrophysiological response (electroretinogram) elicited on isolated retina by a brief light flash. Immediately after the administration of (R)PAF-acether, an irreversible decrease of the electroretinogram b-wave amplitude is observed. This effect is dose dependent (2 X 10(-11) M, 2 X 10(-9) M, 2 X 10(-7) M) and partially inhibited by simultaneous administration of Ginkgolide B (BN 52021; 2 X 10(-5) M). These results suggest the existence of (R)PAF-acether-specific receptors inside the retina. PMID- 3038785 TI - [Changed phosphodiesterase activity of leukocytes in atopic eczema]. AB - Leukocytes of patients with atopic eczema exhibit a variety of biochemical abnormalities, particularly an alteration in cyclic nucleotide metabolism. The basic defect is an elevation of phosphodiesterase activity in these cells, which leads to a diminished modulation of cellular immune function by cAMP. This, in turn, leads to an elevation of IgE synthesis and enhanced release of inflammatory mediators from mast cells and basophils. The phosphodiesterase defect offers new approaches to the treatment of atopic eczema. PMID- 3038787 TI - Synthesis of [11C]SCH 23390 for dopamine D1 receptor studies. AB - The optimal conditions for the synthesis of 11C-labelled SCH 23390 by radio methylation of its desmethyl precursor, SCH 24518, with [11C]iodomethane are described. Isocratic reversed phase HPLC was used for the purification of [11C]SCH 23390. The specific activity range in 30 runs was 10-235 Ci/mmol and average radiochemical yield was 72% based on [11C]iodomethane. Mean synthesis time was 40-60 min from the end of bombardment. Preliminary animal studies indicate that [11C]SCH 23390 would be useful in visualizing D1 receptors in a living brain by positron tomography. PMID- 3038786 TI - A simple method for the quantitation of 14C-whole-body autoradiograms. AB - A simple method for the quantitation of 14C-whole-body autoradiograms by comparative densitometry is described. If processed under the same conditions as the samples of tissues to be investigated, a radioactive blood scale can be used as standard, with the exception of samples from bones, eyes and fat. This method is demonstrated to be simple, accurate, reproducible and precise. PMID- 3038788 TI - Synthesis and biodistribution of [125I]iodo- and [75Se]seleno-ergoline derivatives. AB - (8 beta)-8-([125]Iodomethyl)-6-propylergoline (125I-3) was prepared by refluxing the mesyl analog with Na125I in methyl-ethyl-ketone, followed by HPLC, in a radiochemical yield greater than 70%. [75Se]Selenopergolide (75Se-2) was prepared in 74% yield starting with H75(2) SeO3. The biodistribution studies of the two compounds in male rats show good uptake by the adrenals and the brain. Compound 75Se-2 had higher adrenal uptake and adrenal-to-blood ratios (4.2% dose/g and 70:1) than 125I-3 (3.6% dose/g and 23.8:1) at 15 min post injection. The two compounds had almost equal brain uptake (0.91% dose/g for 75Se-2 and 1.14% dose/g for 125I-3), but 75Se-2 showed higher brain-to-blood ratios (15.2:1 vs 7.3:1) at 15 min post injection. This study indicates that 75Se-2 and 123I-3 may be useful agents for imaging the adrenal and the brain. PMID- 3038789 TI - Measurement of 99Tc in low-level radioactive waste from reactors using 99mTc as a tracer. AB - A radiochemical procedure has been developed for measuring 99Tc in major low level radioactive waste streams from commercial nuclear power facilities which contain numerous potential contaminants. Separation was by alkaline precipitation and anion exchange chromatography; trace amounts of 60Co and other radioactive contaminants were removed by solvent separation. Pure separations were obtained with average radiochemical yield of 47.3 +/- 4.3% determined by a 99mTc tracer. The lower limit of detection for 99Tc was found to be 0.16 pCi/g based on 10-g sample and a 100-min counting time. PMID- 3038790 TI - Beta spectroscopy with liquid scintillation systems: applications in dosimetry of 90Sr and 90Y. AB - We have demonstrated that commercially available liquid scintillation counters are adequate for obtaining useful information about the variation of beta spectral shapes as a function of attenuation. A gel scintillation system that permitted the approximation of point source geometry worked well. The spectral shapes predicted by the Fermi equations were reasonably matched with experimental spectra of sources with geometry close to a point source. For energies in the range of 50-2300 keV, we found sufficient linearity of the energy scale and uniformity of detection limits for the direct output of the instrument to be experimentally useful. The gel scintillation system has been used to directly measure the energy distribution of the beta flux coming out of bone samples and incident on the soft tissue of dogs fed 90Sr-90Y from in utero to 540 days of age. PMID- 3038791 TI - Determination of the oxidation state of 99mTc in complexes with MDP and DTPA. AB - The oxidation state of technetium-99m, reduced by concentrated hydrochloric acid and evaporation until dryness, was determined by radiochromatography. The dry deposits of reduced 99mTc were allowed to react with DTPA and MDP with and without antioxidants present. The formation of 99mTc(IV)-DTPA and 99mTc(IV)-MDP complexes and the influence of the antioxidants gentisic and ascorbic acids were studied by radiochromatography and by biodistributional studies in rabbits. 99mTc(IV)-MDP complexes thus formed were demonstrated to have the same biodistribution in rabbits as 99mTc-MDP complexes formed by a conventional stannous reduction kit technique. PMID- 3038792 TI - Comments on tritium monitoring by liquid-scintillation counting. PMID- 3038794 TI - Elimination of the non-specific binding of avidin to tissue sections. AB - A simple procedure is described for eliminating non-specific staining with avidin peroxidase conjugates. Murine ovaries were embedded in either paraffin wax or epoxy resin and, after blocking endogenous peroxidase activity, were treated with 10 micrograms/ml biotinylated Pisum sativum agglutinin. Avidin-peroxidase conjugates (5 micrograms/ml), diluted in standard 0.05 M tris-buffered saline, pH 7.6, containing 0.139 M NaCl, produced considerable background coloration and intense mast cell staining in controls without the lectin. This background diminished as the ionic strength of the buffer was raised. At 0.125 M Tris buffered saline (containing 0.347 M NaCl) the background was completely unstained, with elimination of all binding to mast cells and only minimal loss of specific lectin binding. PMID- 3038793 TI - Histochemical procedures for the simultaneous visualization of neutral sugars and either sialic acid and its side chain O-acyl variants or O-sulphate ester. I. Methods based upon the selective periodate oxidation of sialic acids. AB - Five new methods, based upon the selective oxidation of sialic acid residues with 0.4 mM periodic acid in approximately 1 M hydrochloric acid at 4 degree C for 1 h (PA), have been devised for the simultaneous visualization of neutral sugars and either sialic acid and its side chain O-acyl variants or O-sulphate ester. In the first of these, the selective periodate oxidation-borohydride reduction saponification-selective periodate oxidation-Thionin Schiff-saponification borohydride reduction-periodic acid-Schiff (PA-Bh-KOH-PA-T-KOH-Bh-PAS) technique, sialic acids with O-acyl substituents at C7, C8 or C9 (or which have two of three side chain O-acyl substituents) stain blue while neutral sugars with periodate sensitive vicinal diols (hexose, 6-deoxyhexose, and N-acetylhexosamine) stain magenta. The second method, the saponification-selective periodate oxidation Thionin Schiff-saponification-borohydride reduction-periodic acid-Schiff (KOH-PA T-KOH-Bh-PAS), stains all sialic acids blue and neutral sugars magenta. In the third procedure, the selective periodate oxidation-Thionin Schiff-borohydride reduction-periodic acid-Schiff-saponification (PA-T-Bh-PAS-KOH) method, sialic acids without side chain substituents (or which have an O-acyl substituent at C7) stain blue and neutral sugars stain magenta. In the fourth method, the saponification-selective periodate oxidation-borohydride reduction-Alcian Blue pH 1.0-periodic acid-Schiff (KOH-PA-Bh-AB1.0-PAS) technique, O-sulphate esters stain aquamarine blue and neutral sugars stain magenta. In all of these techniques mixtures of the components stain in various shades of purple.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3038795 TI - Immunocytochemical study of the intracellular localization of M protein of vesicular stomatitis virus. AB - The purpose of this paper is to describe the immunocytochemical localization of M protein of vesicular stomatitis virus (VSV) in infected cells. Vero cells, MDBK cells, Swiss 3T3 cells, and BHK cells were examined at various times after infection. For immunofluorescent staining, the cells were fixed with PLP fixative and then treated with 0.05% Triton X-100 before incubation with antibodies. Three hours after infection, M protein exhibited diffuse immunostaining throughout the cytoplasm and later accumulated along the cell membrane. The localization of M protein differed from the granular localization of the nucleocapsid N protein of VSV in the cytoplasm. For electron microscopy, the cells were fixed first in a mixture of 2% paraformaldehyde and 0.05% glutaraldehyde and then with PLP fixative, this being followed by treatment with 0.05% saponin. They were then immunostained using the immunoperoxidase method. The M protein was found to be distributed throughout the cytoplasm and later under the cell membrane, especially at virus budding sites. We also used postembedding immunostaining and freeze-fracture immunostaining to avoid the translocation of M protein caused by the detergent treatment. These techniques confirmed our previous results. Our findings are consistent with the view that the M protein of VSV is synthesized on free ribosomes and is then associated with the cell membrane where viral assembly may occur. PMID- 3038796 TI - Regional differences in the uptake of exogenous copper into rat brain after acute treatment with sodium diethyldithiocarbamate. A histochemical and atomic absorption spectrophotometric study. AB - Since sodium diethyldithiocarbamate (SDEDTC) is known to increase the tissue uptake of copper, we have examined its effect on copper accumulation in the rat cerebellum, hypothalamus, parietal cortex and hippocampus by means of atomic absorption spectrophotometry. Acute SDEDTC (1,000 mg/kg i.p.) administration alone did not alter the regional concentration of copper in the cerebellum, hypothalamus and parietal cortex, but significantly increased it in the hippocampus, 5 h after treatment. Copper acetate (5 mg/kg) given i.p. has a stimulatory effect on copper uptake only in the hypothalamus and hippocampus. When copper acetate was administered to rats which were pretreated with SDEDTC, an especially high significant increase in the hippocampal copper level could be observed (approximately 70%), while the enhancement in cerebellar copper concentration was much more lower (approximately 20%), but yet significant. These data suggest that SDEDTC enhances the uptake of exogenous copper in all brain regions examined since the lipophilic SDEDTC-copper complexes easily penetrate the cell membranes. Furthermore, our histochemical findings indicate that--under normal conditions--copper is stored predominantly in glial cells, while following an excessive uptake this metal is also accumulated in neurons (e.g. pyramidal cells of the hippocampus and cortex). PMID- 3038799 TI - [Pulse synchronous vascular tinnitus: radiologic diagnosis and therapy]. AB - Tinnitus synchronous with the pulse is usually caused by vascular anomalies. In our own patient group we found the most frequent cause to be highly vascularised tumours related to the pretrous bone, the most common being glomus tumours. Pulsatile tinnitus however also is a main symptom of dural arterio-venous malformations with shunts into the transverse and sigmoid sinus. Finally pulsatile tinnitus may be caused by venous deformities, possibly combined with anomalies of the bulb of the jugular vein. While clinical methods give valuable clues to the type of pathological findings, subtle radiological investigations are necessary for the final diagnosis, such as high resolution computer tomography and super-selective angiography. There has been substantial progress in recent years due to technical developments leading to improved diagnosis and treatment by interventional radiology. PMID- 3038798 TI - Autoradiographic studies with 3H 1,25 dihydroxyvitamin D3 in thyroid and associated tissues of the neck region. AB - Rats and mice fed a vitamin D-deficient or vitamin D-complete diet were injected with 3H 1,25 (OH)2 vitamin D3. Autoradiograms prepared from cross sections through the neck region revealed nuclear concentration of radioactivity strongest in parathyroid chief cells, occasionally in thyroid follicular epithelial and interfollicular cells, in the epithelium of tubular remnants of the ultimobranchial body, in epithelium of the esophagus, in chondrocytes of tracheal cartilage, and in myoepithelial cells of tracheal glands. In the thyroid, most of the follicle epithelial cells did not show nuclear concentration of radioactivity which occurred only occasionally and predominantly in follicles located in marginal positions. Thyroglobulin in lumina of thyroid follicles contained varying amounts of radioactivity that correspond to the diameter of the follicles, with relatively high amounts in large follicles and little or no radioactivity in small follicles. Competition with excess of unlabeled 1,25 (OH)2 vitamin D3 abolished nuclear radioactivity, but not the radioactivity in the colloid, while 25 (OH) vitamin D3 did not affect either. When a combination of autoradiography and immunohistochemistry was applied, follicular and parafollicular C-cells positive for calcitonin antibodies, did not show nuclear concentration of radioactivity. Tubular remnants of ultimobranchial bodies, however, showed distinct nuclear labeling, but did not stain, or only weakly stain, with antibodies to calcitonin. When 3H 25 (OH) vitamin D3 was injected, no nuclear concentration of radioactivity was noted in any of the tissues.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3038797 TI - Skeletal muscle cell populations. Separation and partial characterization of fibroblast-like cells from embryonic tissue using density centrifugation. AB - Cell suspensions from the breast muscles of 10-day old chicken embryos were separated into non-myogenic, fibroblast-like cell fractions and a mononucleated, myogenic cell fraction by Percoll density centrifugation. Isolated populations were characterized by their morphology in both mass cultures and individual macroscopic clones and by the immunocytochemical detection of skeletal muscle- and smooth muscle-specific proteins in individual cells. Cell populations were also characterized by their protein patterns using sodium dodecyl sulfate/polyacrylamide gel electrophoresis. The less dense, non-myogenic cells comprised 16% of the cells. In culture they were predominantly flattened, stellate cells and gave rise to clones lacking myotubes. These fibroblast-like cells were negative for skeletal muscle myosin or muscle type creatine phosphokinase. Less than 0.1% of these cells demonstrated strong fluorescence when stained with anti-desmin or anti-smooth muscle specific actin. This observation suggested that the vast majority of these cells were not related to vascular smooth muscle cells. Also, over 99% of the non-myogenic cells did not display characteristic properties of endothelial cells. The denser myogenic cell fraction comprised over 80% of the cells and in clonal cultures gave rise to about 70% myogenic clones. An additional 30% of clones from this fraction were non-myogenic indicating heterogeneity in this population. We conclude that Percoll centrifugation can be employed for the isolation of myogenic and non myogenic cell populations directly from the embryonic muscle. Moreover, this procedure allows the direct analysis of cell-specific proteins (e.g., by gel electrophoresis) without the need for cell culturing. The results thus obtained closely reflect the status of the cells in the intact muscle. PMID- 3038801 TI - The role of radiotherapy in the management of adenoid cystic carcinoma of the head and neck. AB - Sixty-eight patients with adenoid cystic carcinoma (ACC) of the head and neck treated by radiation with or without surgery were reviewed. The tumors were located in the major salivary glands in 15 patients and in the minor glands of various sites, mostly in the palate and/or upper gum, in 53. The local control rate was naturally better in the patients with elective postoperative radiation and those with recurrent disease who underwent radiation after repeated surgery for the purpose of tumor reduction than in patients with residual and/or inoperable tumors. For localized tumors in the oral cavity or maxillary sinus, local control was excellent using brachytherapy or "trimodal" combined therapy (radiation, drugs, and minor surgery). The effectiveness of radiation seemed to be more significantly related to the size of the tumor than the dose of radiation. The prognosis of the patients with a predominantly "solid" histologic pattern was apparently worse and presented fulminating progression of the disease. Among patients with a histologically typical cribriform pattern and/or undetermined pattern, although distant metastasis developed frequently, the majority survived a relatively long period without functional or cosmetic disturbance to their normal activities associated with slow progression of the disease. In minor gland tumors, as long as the lesion was localized and a solid histologic pattern was unlikely, radical radiation could be considered an effective choice with the advantage of enhancing the quality of life for the patients. PMID- 3038800 TI - Polymorphisms within the HLA-DRw6 haplotype. III. DQ alpha and DQ beta polymorphism associated with HLA-D. AB - We have studied HLA-DQ encoded antigens from HLA-DRw6 homozygous cells and analyzed the DQ region at the DNA level. HLA-DQ molecules were isolated from EBV transformed B-cell lines and analyzed for DQ alpha and DQ beta polymorphism. From the same set of cells, DNA was isolated and analyzed for RFLP. Polymorphism could be detected by both techniques, i.e., on the protein level and on the DNA. The variation in pI of the DQ alpha and beta chains correlated with the polymorphism as detected by HTC typing, as did the variation in molecular weight of the bands hybridizing to DQ specific cDNA probes; identical patterns were detected for cells of one HLA-D specificity and different patterns for different HLA-D types. Additionally, DQ reactive PLT reagents were raised against DRw6 positive cells. Panel studies revealed that these DQ reactive proliferative T cells can discriminate between the polymorphic DQ antigens on cells with different HLA-D specificities. PMID- 3038803 TI - Testing for feline leukemia. PMID- 3038802 TI - Enhancement of radiation effects by alpha interferon in the treatment of small cell carcinoma of the lung. AB - The effects on lung tissue and tumor of natural human alpha interferon (IFN) and radiotherapy were investigated in a multimodality treatment program for selected patients with small cell carcinoma of the lung (SCLC). Interferon was given first as a single agent, then concomitantly with radiotherapy to 12 previously untreated patients with limited disease. At disease progression outside the chest, interferon was discontinued and combination chemotherapy was initiated. In the first series, 7 patients received a high interferon induction dose (800 X 10(6) IU i.v. over 5 days) followed by low-dose maintenance therapy (6 X 10(6) IU i.m. TIW), median total dose 1380 X 10(6) IU (range 794-2074). At local progression, split-course radiotherapy, 55 Gy/20 F/7 wk, was added to interferon therapy. In the second series, 5 patients received low-dose interferon from the start (6 X 10(6) IU i.m. daily) combined with twice-a-day fractionated radiotherapy 44 Gy/40 F/4 wk. Median total dose of interferon in this series was 698 X 10(6) IU (range 354-828). Tumor response and normal tissue reactions were evaluated by monthly chest X rays, 3-monthly CT scans, restaging bronchoscopies and by serial respiratory function tests. Autopsy specimens from both lungs within and outside the radiation field were systematically evaluated when available. After the completion of radiotherapy, there were 4/7 CR in the high dose IFN group compared to 3/5 CR in the low-dose IFN group. Rapid shrinkage of huge tumor masses was observed. At 2 months post radiotherapy radiological grade III fibrosis occurred in 4/7 patients in the high-dose and 1/5 patients in the low-dose group. Lung function studies showed a significant decrease in diffusing capacity and in lung volumes. Seven patients died within 12 months from start of interferon treatment, one of them from treatment complication. At autopsy the tumor area was in most cases replaced by severe fibrosis. Outside the radiation field lung fibrosis was mild. Our results suggest enhancement of radiation effect by interferon with a possible dose and/or schedule dependence of interferon and radiotherapy and call for more clinical studies of IFN and radiotherapy in combination. PMID- 3038804 TI - Bovine viral diarrhea virus: a review. PMID- 3038805 TI - Identification of ovine adenovirus types five and six in an epizootic of respiratory tract disease in recently weaned lambs. AB - Eight hundred fourteen Polypay-cross lambs, 32 to 35 days old, were housed in an isolation barn under semiconfinement conditions. The lambs were observed twice daily for clinical signs of respiratory tract disease. During the 8-week observation period, an epizootic of respiratory tract disease developed, in which the combined morbidity and mortality was approximately 13%. The overall mortality was 4.1%, of which 57.6% was attributable to pneumonia. Pasteurella haemolytica was isolated from 11 (58%) of the 19 lung specimens obtained at necropsy. Serotyping revealed the presence of multiple serotypes: A1, A2, A5, A6, A9, and A12. Viruses with properties consistent with ovine adenovirus were isolated from 36% of the lung specimens. Five of the isolates were selected to determine their antigenic serotype. Results of virus neutralization testing indicated that in this group of lambs, there was concurrent infection with Mastadenovirus ovi type 5 and type 6. PMID- 3038806 TI - Status of equine viral arteritis in Kentucky, 1985. AB - Clinical cases of equine arteritis virus infection have not been diagnosed in Kentucky since 1984, and there has been no indication that any of the horses involved in the 1984 epizootic have since been responsible for spread of the disease to horses in other states or other countries. Cases of abortion caused by naturally acquired infection with this virus have not been confirmed in 1984 or 1985. Neither field nor vaccine strains of equine arteritis virus have been shown to induce teratologic abnormalities or the carrier state in foals born to infected or vaccinated mares. The carrier stallion appears to have played a major epidemiologic role in the dissemination and perpetuation of the virus. A commercial modified live equine viral arteritis vaccine was found to be safe and efficacious for stallions and mares. The disease can be controlled by immunizing the stallion population and by restricting the breeding of equine arteritis virus shedding stallions to vaccinated or seropositive mares, followed by an appropriate period of isolation from other nonvaccinated Equidae. PMID- 3038808 TI - Cyclic hematopoiesis associated with feline leukemia virus infection in two cats. AB - Cyclic oscillations in the numbers of blood elements were detected in 2 cats with FeLV infection. Periodic neutropenia, followed by a return to normal neutrophil numbers, was detected in both cats. The mean cycle duration was 11.8 days, with a range of 8 to 14 days. Just before the return of normal neutrophil numbers, monocytosis developed. In 1 cat, cyclic variations in the number of reticulocytes and platelets also were detected. Bone marrow aspirates obtained during periods of neutropenia had a predominance of progranulocytes in the myeloid cell line. myeloid hyperplasia, with numerous segmented neutrophils, was seen in bone marrow aspirates obtained during periods of normal neutrophil numbers. Oral administration of prednisolone resulted in cessation of the cyclic oscillations of blood elements in 1 cat. Cyclic hematopoiesis appeared to be another non neoplastic manifestation of FeLV infection. PMID- 3038807 TI - Correction of metabolic acidosis in diarrheal calves by oral administration of electrolyte solutions with or without bicarbonate. AB - Acid-base balance was evaluated in calves with experimentally induced viral diarrhea. When blood pH decreased to less than 7.200, calves were assigned to treatment groups and fed milk replacer, electrolyte solution without bicarbonate, or electrolyte solution containing bicarbonate. Calves in the electrolyte treatment groups had lower mortality (P less than 0.05), were better hydrated (P less than 0.05), and were less acidotic (P less than 0.05) than calves fed milk replacer. Bicarbonate-containing electrolyte solution restored acid-base balance (P less than 0.05) and corrected depression better (P less than 0.05) than electrolyte solution that did not contain bicarbonate. Both electrolyte solutions were equally good at correcting dehydration. PMID- 3038809 TI - Hypocalcemia associated with administration of sodium bicarbonate for salicylate intoxication in a cat. AB - Hypocalcemia was believed to be the result of aggressive sodium bicarbonate treatment for a cat with aspirin intoxication. The cat developed progressive neuromuscular tetany during the bicarbonate treatment and was determined to have a low serum ionized calcium value. PMID- 3038810 TI - Myc genes linked to viral promoters can collaborate with a transformation deficient simian virus 40 early gene mutant to transform established rat cells. AB - Cotransfection of a transformation-deficient and immortalization-positive SV40 early gene mutant with various constructs of viral promoter-linked myc genes induced dense foci in 3Y1 cells, an established cell line derived from rat embryo fibroblasts, whereas transfections with either species alone did not. Four cell clones were isolated from colonies grown in soft agar medium, and found to contain at least a single copy of both DNA species and to express both sequences at relatively high levels. These results indicate that the two immortalizing genes used in the present study collaborate to transform 3Y1 cells. PMID- 3038811 TI - Trend of lung cancers in the National Cancer Center of Japan and comparison with that of Japanese pathological autopsy records. AB - Trends in the histologic types of lung cancers during 1966-1985 by 5-year intervals were analyzed by using 4,419 cases in the National Cancer Center Hospital, Japan. Histologically confirmed cases increased to 76.6% in the last 5 year interval. The proportions of histological types in males have remained relatively unchanged; about 40% squamous cell carcinoma, one-third adenocarcinoma, and about 10% each of large cell carcinoma, small cell carcinoma and other types. In females, an increase of adenocarcinoma from 52% to 69.1% and a decrease of squamous cell carcinoma from 21% to 11.4% were noteworthy, and the other three types amounted to less than 10% each. These trends were compared with those obtained from the Annals of Pathological Autopsy Records in Japan during 1974-1983, which contained 26,844 lung cancer cases; the results were similar except for an increase of small cell carcinoma. The recent increase of small cell carcinoma was considered to be due to a change in diagnostic categorization, associated with a decreased rate of undifferentiated carcinoma. However, the increase of small cell carcinoma as well as adenocarcinoma in younger patients requires further study from both demographic and etiological viewpoints. PMID- 3038812 TI - Breakpoints in Philadelphia chromosome (Ph1)-positive leukemias. AB - We studied chromosome 22 breakpoints in 24 Philadelphia (Ph1)-positive leukemias. Nineteen of 21 patients with chronic myelogenous leukemia (CML) possessed a chromosomal break within the 5.8 kilobase (kb) breakpoint cluster region (bcr). Furthermore, in one of three cases of acute lymphocytic leukemia (ALL), we found a chromosomal rearrangement in the bcr locus. No chromosomal rearrangements were found in two cases of CML and two cases of ALL using several restriction enzymes. The bcr rearrangement is highly specific for CML. Further analyses will be necessary to determine whether some cases without bcr rearrangement have another specific locus of rearrangement. Two of three cases of CML with blastic crisis had rearrangement of the immunoglobulin gene and T-cell receptor gene. One CML patient with blastic crisis, who achieved complete remission but relapsed thereafter, was found to have the same immunoglobulin gene rearrangement in the blastic crisis and relapse phases, suggesting that the same clone was involved in both crisis and relapse. PMID- 3038813 TI - A sandwich enzyme immunoassay of an adenocarcinoma-associated antigen, YH206, in cancer sera. AB - A sandwich enzyme immunoassay was established to measure an adenocarcinoma associated antigen (antigen YH206) detected by monoclonal antibody YH206. Levels of antigen YH206 exceeding the cut-off value (25 U/ml) were found in the following percentages of 163 patients with various cancers: stomach cancer 37.2%, colon cancer 14.8%, pancreas cancer 43.3%, common bile duct cancer 28.6%. In contrast, only one out of 33 (3.0%) healthy donors and 7 of 104 (6.7%) patients with benign diseases had slightly elevated levels of the antigen. As regards the relationship between antigen YH206 levels and clinical stages, abnormally high levels of the antigen were found in the following percentages of 29 patients with stomach cancer: stage I 16.7%, II 0%, III 42.9% and IV 54.5%. Serial monitoring of antigen YH206 in two patients with cancer revealed that the level of the antigen increased as the disease progressed. It was also found that perchloric acid treatment of serum might be useful to decrease any false-positive reactions. PMID- 3038814 TI - Sperm-bound amidase activity as a marker for acrosomal integrity in bull spermatozoa. AB - An assay for sperm-bound amidase activity was validated using bovine spermatozoa and N-benzoyl-DL-arginine p-nitroanilide as substrate. The assay had intra- and interassay coefficients of variations of 5 and 12%, respectively. It is an inexpensive, simple and rapid assay since 100 samples can be evaluated in 2 hours and it requires only 4 X 10(6) spermatozoa per sample. Sperm-bound amidase activity was proportional (r = 0.95) to the percentage of spermatozoa with an intact acrosome, as determined by differential interference-contrast microscopy. A change of five percentage units in the incidence of damaged spermatozoa was detectable. Using this procedure, assessment of sperm-bound amidase activity is therefore a sensitive and efficient means of evaluating acrosomal integrity. PMID- 3038815 TI - Effect of adrenal steroids on testosterone and luteinizing hormone secretion in the ram. AB - The effects of adrenal steroids on testosterone and LH secretion and changes in serum cortisol levels in response to treatments were studied in the ram. Acute administration of synthetic ACTH (10 micrograms/kg BW) elevated (P less than 0.01) serum cortisol and transiently suppressed (P less than 0.05) serum testosterone and LH. Acute dexamethasone treatment suppressed (P less than 0.01) serum cortisol, testosterone and LH. Administration of vehicle had no effect (P greater than 0.10) on serum hormone levels. These data support the contention that adrenal steroids inhibit testicular endocrine function indirectly by acting at the hypothalamic or pituitary level because both ACTH and dexamethasone treatments suppressed serum LH. To differentiate between hypothalamic and pituitary sites of action, the pituitary and testicular responses to an LHRH challenge (100 micrograms) were examined in rams chronically treated with dexamethasone (5 mg i.m., twice daily for 5 days). This treatment regimen suppressed (P less than 0.01) serum cortisol levels. Compared with controls, basal testosterone levels were suppressed (P less than 0.05) in dexamethasone treated rams; however, no effect (P greater than 0.10) on the magnitude of the testosterone response to LHRH or on either basal or LHRH-stimulated LH secretion was observed. Thus, although a direct testicular effect cannot be eliminated, these data suggest that, in the ram, adrenal steroids inhibit testicular endocrine function by action at the level of the hypothalamus. PMID- 3038816 TI - Staurosporine inhibits tyrosine-specific protein kinase activity of Rous sarcoma virus transforming protein p60. PMID- 3038817 TI - Studies on antiviral agents. V. Synthesis and in vitro antiviral activity of new aminoglycoside derivatives having palmitoyl group. AB - The synthesis and antiviral activity of various aminoglycoside derivatives having a palmitoyl group are described. All of these aminoglycoside derivatives exhibited almost the same excellent antiviral activity against herpes simplex virus type I and influenza virus. PMID- 3038818 TI - Polycations which disorganize the outer membrane inhibit conjugation in Escherichia coli. AB - Outer membrane disorganizing polycation, polymyxin B nonapeptide (PMBN), reduced drastically the production of recombinants when present at sub-MIC concentrations during F'-mediated Escherichia coli conjugation. The decrease of recombinants was accompanied by a less marked decrease of viability in the recipient population in a manner resembling lethal zygosis. No reduction was seen when either donor or recipient was grown in the same PMBN concentration, washed and resuspended to PMBN-free medium before mating. The same concentration of outer membrane disorganizing polycations of higher bactericidal activity (protamine and polylysine) caused only a moderate reduction in transconjugant frequency when present during mating. Spermine and tetralysine, which are not effective disorganizers of the outer membrane, did not reduce the recombinant frequency or the viability of the recipients. PMID- 3038819 TI - Ontogenetic approach to cellular localization of neurotransmitters in the chick vestibule. AB - Gamma-aminobutyric acid (GABA) and acetylcholine (Ach) have been implicated in afferent and efferent neurotransmission, respectively, in the vestibular sensory periphery. Assuming that glutamate decarboxylase (GAD), the GABA-synthesizing enzyme, and choline acetyltransferase (ChAT), the enzyme for synthesis of acetylcholine, are located in distinct cell types of the inner ear whose maturation occurs at different times during ontogenesis, we measured these enzymes in the ampullary cristae of embryonic chicks at different stages of development. By making these measurements in parallel with electron-microscopic studies of the different cell elements of the chick vestibular sensory periphery, we found that the values of GAD activity were nearly the same from the earliest stage studied, i.e., the 13th day of ontogeny to day 18 of embryonic development, paralleling the morphologically mature appearance of the hair cells and their afferent synapses. A slight increase in enzymatic activity from day 19 of ontogeny to one day after hatching corresponded to a rise in the number of afferent synapses. In contrast, ChAT activity was practically undetectable up to day 17 of embryonic development, but rose suddenly on the 19th day, reaching 1 day-old levels by day 20 of ontogenesis in coincidence with an elevation in the number of well-developed efferent boutons. These results are in accord with the localization of GAD in the sensory cells and a localization of ChAT in the efferent nerve endings. These findings suggest that GABA and Ach are the respective neuromediators for the afferent and efferent systems. PMID- 3038820 TI - Morphometric analysis and fine structure of the vestibular epithelium of aged C57BL/6NNia mice. AB - The vestibular organs of young and very old C57BL/6NNia (B6) mice were compared by light and electron microscopy. Hair cell density decreased an average of 14% in the utricle, 19% in the saccule and posterior crista, 23% in the horizontal crista, and 24% in the anterior crista. Hair cell size remained the same throughout the mouse's life span as did the ratio of Type I to Type II hair cells. The most apparent sign of advanced age was dense inclusions found in sensory and supporting cells. Although small inclusions were present at five weeks, by 29 months, additional, larger forms appeared. An unusual melanin-like form was characteristic of old Type I hair cells. Synaptic morphology and synaptic bodies were well preserved even in very old B6 mice. Elongated bars were common in Type I hair cells and spheroid synaptic bodies were the most common form in Type II hair cells. Large clusters of synaptic bodies occurring in both young and old mice were seen only in Type I hair cells. Although the B6 strain suffers from genetically determined early cochlear degeneration, it does not experience early degeneration of the peripheral vestibular organs. PMID- 3038821 TI - Evaluation of corn fiber, cottonseed hulls, oat hulls and soybean hulls as roughage sources for ruminants. AB - An in situ trial (randomized complete block design) using cows, and a site and extent of digestion trial (Latin square design) using sheep were conducted to study the potential of corn fiber (CF), cottonseed hulls (CSH), oat hulls (OH) and soybean hulls (SH) as roughage sources for ruminants. Two feedlot trials with steers and one with lambs (completely randomized design with factorial arrangements of treatments) were conducted to study the potential of CF and SH as energy supplements relative to corn. In situ rate of ruminal dry matter (DM) disappearance (3 to 36 h) and extent of DM disappearance (36 h) indicated that CF and SH were more fermentable in the rumen compared with OH or CSH, with SH being the most fermentable. Total tract digestibilities of DM, organic matter (OM), neutral detergent fiber (NDF) and acid detergent fiber (ADF) were above 70% for sheep fed CF and SH diets, and were 50% or less for sheep fed OH and CSH diets. A ranking of by-products in terms of nutritive value followed the trend: CF greater than SH greater than OH greater than CSH. Lamb feedlot trial data showed that CF was of similar nutritive value to corn and of higher nutritive value than SH at the 50% level of supplementation. Corn-fed lambs responded better than CF- or SH fed lambs at the 70% level of supplementation. Data from steer feedlot trials showed that CF was of similar quality to corn and of higher quality than SH. Dramatic differences exist in by-product feed utilization by ruminants. All by products tested appeared to have some usefulness as dietary components. PMID- 3038822 TI - Trona and sodium bicarbonate in beef cattle diets: effects on site and extent of digestion. AB - Six yearling Hereford X Angus steers (avg 272 kg), each with ruminal, duodenal and ileal cannulas were used in a 6 X 6 Latin-square metabolism trial to evaluate the impact of NaHCO3 and trona (a ground, nonrefined ore with chemical composition NaHCO3-Na2CO3-2H2O) on site and extent of digestion of nutrients in the digestive tract. The diets were 50:50 or 90:10 (cracked corn-based concentrate:cottonseed hulls) with no buffer, 1% NaHCO3, or 1% trona. Intake, across all treatments, averaged 2.4% of body weight. Dry matter (DM) and starch digestibility (via indigestible acid detergent fiber) before the duodenum was decreased (P less than .10) with trona in the 50:50 diet. Digestibility of DM, crude protein and starch before the ileum were greater (P less than .05) in the 90:10 diet vs 50:50 diet. Total tract digestibility was similar across buffer treatments in the 90:10 diet. Addition of NaHCO3 increased (P less than .05) digestibility of dry matter and cell solubles in the 50:50 diet. Organic matter and crude protein digestibility were also increased (P less than .10) with NaHCO3. Apparent crude protein and cell solubles digestibility were greater (P less than .10) with trona than NaHCO3 in the 50:50 diet. This trial indicates that buffers provide overall enhancement of diet digestibility in mixed grain/roughage diets. PMID- 3038823 TI - Trona and sodium bicarbonate in beef cattle diets: effects on pH and volatile fatty acid concentrations. AB - A study was conducted to evaluate the effectiveness of NaHCO3 and trona in beef cattle diets. Trace element (n = 28) analysis revealed no toxicological or safety concerns with the use of trona. Trona was more (P less than .05) soluble in ruminal fluid than Na2CO3, and NaHCO3 and had greater (P less than .05) buffering capacity (9.6 meq/g) than NaHCO3 (6.1 meq/g) but less (P less than .05) than Na2CO3 (11.1 meq/g). Calcium carbonate was insoluble and did not buffer ruminal fluid. Six yearling (avg 272 kg) Hereford X Angus steers, each with ruminal, duodenal and ileal cannulas, were fed 50:50 (cracked corn-based concentrate:cottonseed hulls) or 90:10 concentrate diets with no buffer, 1% NaHCO3 or with 1% trona. Intake, across all treatments, averaged 2.4% of body weight. Propionate (mmol/liter) increased (17.6 vs 13.5; P less than .05) and butyrate decreased (3.5 vs 5.2; P less than .05) with trona in the 90:10 diet as compared with no buffer. Propionate (16.8) increased (P less than .05) with NaHCO3 in the 90:10 diet. Average ruminal pH was greater (P less than .05) in 90:10 diets with trona or NaHCO3 than with no buffer (5.61, 5.61 vs 5.55); duodenal pH was greater (P less than .01) with trona than with no buffer (2.66 vs 2.55). Trona reduced ruminal pH-hours (P less than .05) and pH-area (P less than .12; time and area below mean pH of control) below control for both concentrate levels.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3038824 TI - Efficacy of fluconazole (UK-49,858) against experimental aspergillosis and cryptococcosis in mice. AB - The efficacy of fluconazole, a new bis-triazole antifungal agent, was compared with that of orally administered ketoconazole and parenterally administered amphotericin B against aspergillus and cryptococcus infections in mice. Fluconazole was 5-20-fold more active than ketoconazole against systemic aspergillosis and against systemic, intracranial and pulmonary cryptococcosis but was less active than amphotericin B. PMID- 3038825 TI - Alpha-adrenergic agents have little effect during air embolism lung injury in awake sheep. AB - Since it is not clear whether alpha-adrenergic receptors can modulate lung microvascular liquid and protein leakiness, we studied the effects of alpha adrenergic receptor stimulation or blockade on lung filtration under base-line conditions and during the acute lung injury caused by a 4-h infusion of venous air emboli in six unanesthetized, chronically instrumented sheep with lung lymph fistulas. During the experiments we continuously infused the alpha-adrenergic receptor agonist phenylephrine hydrochloride (1.0 microgram X kg-1 X min-1 iv) or the alpha-adrenergic receptor antagonist phentolamine mesylate (1.0 mg X kg-1 X min-1 iv), and we measured pulmonary vascular pressures, cardiac output, lung lymph flow, and the lymph-to-plasma protein concentration ratio. During air embolism, alpha-receptor stimulation increased pulmonary vascular resistance and decreased lung lymph flow by 25%; alpha-receptor blockade had the opposite effects. During recovery, neither agent significantly affected pulmonary hemodynamics or lymph flow. Our results indicate that alpha-adrenergic receptors are active during air embolism and modulate pulmonary filtration by causing arteriolar constriction, which reduces the surface area or the perfusion pressure in the pulmonary microvascular bed. They may also affect venous smooth muscle tone. We found no evidence that alpha-adrenergic receptors modulate lung microvascular liquid or protein leakiness directly. PMID- 3038826 TI - Platelet aggregation increases cholinergic neurotransmission in canine airway. AB - To determine whether thromboxane A2 released from aggregating platelets increases the contractile response of airway smooth muscle to cholinergic nerve stimulation and, if so, what the mechanism of action is, we studied in vitro bronchial segments from dogs under isometric conditions. The contractile responses to electrical field stimulation at 30 s and 1 min after the addition of autologous platelets were increased by 11.1 +/- 3.2 (SD) and 20.7 +/- 5.4%, respectively, and were accompanied by the release of thromboxane A2. These effects were inhibited either by pretreatment of platelets with indomethacin or by addition of the thromboxane A2 receptor antagonist SQ 29548. Likewise, the thromboxane A2 mimetic U 46619, in subthreshold doses (i.e., insufficient to increase base-line tension), increased electrical field stimulation-induced contraction by 18.7 +/- 4.8%. The increase was greater in the presence of a concentration of physostigmine that did not cause spontaneous contraction and was blocked by SQ 29548 but not by hexamethonium or by phentolamine. Methacholine-induced contractions were unaffected by U 46619. These results indicate that aggregating platelets, by releasing thromboxane A2, increase the airway contractile response to neural stimulation probably by the accelerated release of acetylcholine. PMID- 3038827 TI - Respiratory depressant effects of GABA alpha- and beta-receptor agonists in the cat. AB - The aim of this study was to evaluate the cardiorespiratory effects of intravenously administered gamma-aminobutyric acid (GABA) alpha-(4,5,6,7 tetrahydroisoxazolo[5,4-c]pyridin-3-ol, THIP) and beta-(baclofen) receptor agonists and to locate the site of action of these drugs in the brain. THIP and baclofen were administered to alpha-chloralose-anesthetized cats while minute ventilation (VE), arterial blood pressure (AP), and heart rate were monitored. THIP, in doses of 0.5 to 2 mg/kg decreased VE, tidal volume (VT), and AP. No changes in respiratory rate (f) or inspiratory (TI) or expiratory (TE) duration were observed. Baclofen, in doses of 0.5 to 4 mg/kg, decreased VE, f, and AP. VT and TI increased and an "apneustic" breathing pattern was seen. THIP (9.5 micrograms), applied bilaterally to the glycine-sensitive area of the ventral medulla, reproduced the effects seen with intravenous administration. Application of 10 micrograms of bicuculline bilaterally to this area reversed the effects of intravenous THIP but not those of baclofen. Baclofen (5.6-56 micrograms), administered by the intracisternal route, produced the same respiratory effects seen with intravenous administration. We conclude that activation of GABA alpha- and beta-receptors produces cardiorespiratory depression. However, this is accomplished by different mechanisms and by actions exerted at different central nervous system sites. PMID- 3038828 TI - Degradation and reutilization of alveolar phosphatidylcholine by rat lungs. AB - We have shown previously that phospholipids instilled through the trachea are removed from the air spaces in isolated rat lungs by a process that is stimulated by beta-adrenergic agonists. In this study, we evaluated the fate of radiolabeled lipid vesicles [50% [3H]dipalmitoyl phosphatidylcholine (DPPC), 25% phosphatidylcholine (PC), 15% cholesterol, and 10% phosphatidylglycerol (PG)]. Vesicles were instilled through the trachea of anesthetized rats, and the lungs removed for perfusion. The percent of instilled 3H that could not be removed from lungs by extensive lung lavage increased progressively; at 3 h this fraction was 25.8 +/- 0.63% (mean +/- SE; n = 8). The percent of dpm in the lung homogenate accounted for by PC decreased progressively while dpm in lyso-PC, unsaturated PC, and aqueous soluble metabolites [choline, choline phosphate, glycerophosphorycholine, and cytidine 5'-diphosphate (CDP) choline (CDP-choline) increased. The dpm in microsomal and lamellar body fractions isolated from lung homogenates also increased progressively with time of perfusion. The presence of 8-bromoadenosine 3',5'-cyclic monophosphate (8-BrcAMP) significantly stimulated both uptake of DPPC and the appearance of radioactivity in metabolites and subcellular organelles. This effect of 8-BrcAMP was not due to stimulation of phospholipase A activity. These results indicate that exogenous phospholipids instilled into the air spaces of rat lungs are internalized and degraded by a process that is stimulated by cAMP.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3038829 TI - Effects of iron deficiency and training on mitochondrial enzymes in skeletal muscle. AB - We measured mitochondrial enzyme activities in skeletal muscle under conditions of iron deficiency and endurance training to assess the effects of these interventions on the contents and proportions of non-iron-containing and iron dependent enzymes and proteins. Male Sprague-Dawley rats, 21 days of age, received a diet containing either 6 (iron deficient) or 50 mg iron/kg diet (iron sufficient). At 35 days of age animals were subdivided into sedentary and endurance training groups (running at 0.7 mph, 0% grade, 45 min/day, 6 days/wk). By 70 days of age, iron deficiency had decreased gastrocnemius muscle cytochrome c by 62% in sedentary animals. In contrast, the activities of tricarboxylic acid cycle enzymes were increased, remained unchanged or were slightly decreased, indicating that iron deficiency markedly altered mitochondrial composition. Endurance training increased cytochrome c (35%), tricarboxylic acid cycle enzymes (approximately 15%), and manganese superoxide dismutase (33%) in iron-deficient rats, whereas the same exercise regimen had no effect on the skeletal muscle of iron-sufficient animals. The interactive effect of dietary iron deficiency and mild exercise on mitochondrial enzymes suggests that adaptation to a training stimulus is, to some extent, geared to the relationship between the energy demand of exercise and the capacity for O2 transport and utilization. PMID- 3038830 TI - Myocarditis and cardiomyopathy in the dog and cat. AB - Myocarditis and cardiomyopathy were diagnosed in 36 dogs from 11 litters, and myoendocarditis and restrictive cardiomyopathy were diagnosed in 51 cats. Most of the dogs and cats died unexpectedly. Spontaneous parvoviral infection in the dogs caused acute (myocytolysis with presence of intranuclear inclusion bodies in the myocytes), subacute (inflammatory reaction and myocytolysis), and chronic (fibrosis and myocytolysis) myocarditis, which led to extensive myocardial fibrosis and abnormality of the myocytes, similar to dilated cardiomyopathy in man. Spontaneous acute, subacute, and chronic myoendocarditis in the cats led to granulation, extensive fibrosis, and necrosis of the myoendocardium, i.e., like restrictive cardiomyopathy which occurs in man without eosinophilia. Thus, the dog and cat are important animal models of primary myocardial disease. PMID- 3038831 TI - Myocardial lesions by Coxsackie virus B3 and cytomegalovirus infection in infants. AB - Immunofluorescent and electron-microscopic studies were performed to determine the distribution of viral antigens and particles and to clarify the relationship to myocardial lesions in two autopsy cases with generalized infection of Coxsackie virus B3 (CVB3) or cytomegalovirus (CMV). Case 1 was a full-term newborn female infant, without any congenital anomalies, who died of cardiac failure 10 days after birth. CVB3 was isolated from the blood before death. Necrosis of the muscle fibers was observed, frequently accompanying calcification. Numerous histiocytes and a few lymphocytes and neutrophils had infiltrated in and around the necrotic areas. Immunofluorescent study (IF) revealed CVB3 antigen in the muscle fibers and vascular endothelial cells. Case 2 was a female infant, born at 28 weeks of gestation, who died of fatal arrhythmia 50 days after birth. The infant had hemocephalus and a history of idiopathic respiratory distress and underwent an operation for patent ductus arteriosus. Cytomegalic cells were frequently found in the vascular endothelial cells in the myocardium and occasionally in muscle fibers. IF showed the presence of CMV antigen in both endothelial cells and muscle fibers. CVB3 and CMV antigens were detected predominantly in vascular endothelial cells rather than in the muscle fibers. Blood flow disturbance due to endothelial damage is a cause of the myocardial lesion in addition to the direct invasion of the muscle fibers by the virus. PMID- 3038832 TI - Cell-mediated immunity in Coxsackie B3 virus myocarditis in mice--in situ characterization by monoclonal antibody of mononuclear cell infiltrates. AB - This light- and electron-microscopic study using monoclonal antibody and anti immunoglobulin antibodies in murine Coxsackie B3 virus myocarditis provides an immunohistochemical demonstration of surface antigens of lymphocytes. On the 7th and 9th days after inoculation, many necrotic cardiocytes were surrounded by numerous cellular infiltrates, in which macrophages and T lymphocytes predominated, whereas immunoglobulin-bearing B lymphocytes represented a minority. Immuno-electron microscopy showed some T lymphocytes in close contact with other lymphocytes, macrophages, and the sarcolemma of cardiocytes. After the 30th day, significant numbers of T lymphocytes and macrophages were still identifiable in and around the fibrotic foci. Our study suggests that cell mediated immunity plays a protective role by lysing and scavenging virus-infected cardiocytes and cell debris at least in the early stage of myocarditis. The residual T lymphocytes in the chronic stage suggest their involvement in sustained cardiocyte injury. PMID- 3038833 TI - Infections and dilated cardiomyopathy in Nigeria. AB - The relationship of infection to dilated cardiomyopathy is reviewed on the basis of 200 patients seen at University College Hospital, Ibadan. Evidence of infection with Toxoplasma and Coxsackie B viruses is presented. Clinically detectable myocarditis is rare in children, and the preponderance of dilated cardiomyopathy is in patients above the age of 30 years, possibly because there is a long latent period between the initial infection and the development of frank cardiomyopathy. This paper concluded that infections are probably the most important cause of dilated cardiomyopathy in Nigeria. PMID- 3038834 TI - Isolation of Echovirus type 11 and Enterovirus type 71 in a day care winter outbreak. PMID- 3038835 TI - Determination of vitamin D3 in liquid multivitamin preparation, using reverse phase, solid phase extraction liquid chromatography. AB - A method is described for the determination of vitamin D3 in a liquid multivitamin preparation by liquid chromatography. Samples are purified on a disposable reverse phase extraction (SPE) column with a mobile phase of methanol 2-propanol (97 + 3) and are analyzed on a Zorbax ODS (5 micron) column with an acetonitrile-2-propanol-water (90 + 8 + 2) solvent system. Vitamin D3 is completely resolved from other interfering compounds within approximately 21 min and is detected with a UV detector at 254 nm. A mean of 98.5% of theory with a coefficient of variation of 3.8% was found for determination of vitamin D3 in a commercial preparation. PMID- 3038837 TI - Comparative study of respiratory changes in nitric acid plant workers. PMID- 3038836 TI - Brain opioids and autism: an updated analysis of possible linkages. AB - Considerable clinical evidence suggests that autistic children lack the normal ability or desire to engage others socially, as indicated by their poor social skills and inappropriate use of language for communicative purposes. Specifically, these children seem to lack normal amounts of social-emotional interest in other people, leading perhaps to a decreased initiative to communicate. This paper summarizes experimental evidence supporting a neurological theory, which posits that autism, at least partially, represents in the brain, such as brain opioids. These substances modulate social-emotional processes, and the possibility that blockade of opioid activity in the brain may be therapeutic for early childhood autism is discussed. PMID- 3038838 TI - Regulation of the Escherichia coli uvrD gene in vivo. AB - The roles of two putative promoter sequences, P1 and P2, and a potential antiterminator sequence found in the uvrD control region were examined in vivo. Constitutive and SOS-induced levels of uvrD mRNA were determined by S1 mapping, and it was shown that the majority of uvrD transcripts are from P1, while P2 plays only a minor role. A series of increasing deletions from the 5' end of the uvrD gene was used to assay transcription in the promoterless vector pKO-1. Loss of just the -35 region of P1 was sufficient to switch off detectable transcription from both P1 and P2. Disruption of the antiterminator by site specific mutagenesis had no effect on constitutive levels of transcription, but led to a significant increase over wild-type levels following SOS induction. This suggests that the attenuator comes into play following DNA damage to moderate the increase in UvrD protein synthesis. PMID- 3038839 TI - Phosphate regulation of gene expression in Vibrio parahaemolyticus. AB - The synthesis of a major outer membrane protein, OmpP, in Vibrio parahaemolyticus was induced by growth in media deficient in phosphate. The gene, ompP, encoding this protein was cloned. Synthesis of OmpP in Escherichia coli was regulated by the availability of phosphate, and this control required the function of pho regulatory genes of E. coli. Analysis of gene fusion strains constructed by mutagenesis with transposon mini-Mulux revealed that ompP was transcriptionally regulated in V. parahaemolyticus. Impaired growth of a strain with an ompP defect was observed in media which contained large linear polyphosphates as the phosphate source. This and other evidence suggested that OmpP functions as a porin channel for the entry of phosphate into the cell. A number of other proteins or activities were induced by phosphate limitation including hemolysin, phospholipase C, and phosphatase activities. A regulatory locus controlling expression of phosphate-regulated genes was identified and cloned. This regulatory locus cloned from V. parahaemolyticus was shown to complement E. coli strains with defects in pho regulatory genes. PMID- 3038840 TI - Transposition of Tn4551 in Bacteroides fragilis: identification and properties of a new transposon from Bacteroides spp. AB - Tn4551, a clindamycin resistance (Ccr) transposon from the R plasmid pBI136, was cloned onto an Escherichia coli-Bacteroides shuttle vector which could replicate normally in E. coli but was maintained unstably in Bacteroides fragilis. To aid in cloning and to ensure maintenance of Tn4551 in E. coli, a kanamycin resistance determinant (Kmr) was inserted in the transposon. The transposon-bearing shuttle vector pFD197 was transformed into B. fragilis 638, and putative insertions of Tn4551::Kmr were identified by screening for resistance to clindamycin and plasmid content. Southern hybridization analyses were used to verify integration of the transposon in the B. fragilis chromosome, and the frequency of insertion was estimated at 7.8 X 10(-5) events per generation. In 57% of the isolates tested a second integration event also occurred. This second insertion apparently involved just a single copy of the 1.2-kilobase repeat sequence which flanks the transposon. In addition, Tn4551::Kmr appeared to function as a transposon in E. coli. Evidence for this was obtained by the isolation of transposon insertions into the bacteriophage P1 genome. Finally, the transposon vector, pFD197, could be mobilized to other B. fragilis strains in which transposition was detected. Mobilization from the strain 638 background was via a conjugation like process, but occurred in the absence of known conjugative elements or other detectable plasmids. This result suggested the presence of a host-encoded transfer system in this B. fragilis strain. PMID- 3038841 TI - Transfer functions of the Streptococcus faecalis plasmid pAD1: organization of plasmid DNA encoding response to sex pheromone. AB - The conjugative plasmid pAD1 (59.6 kilobases) of Streptococcus faecalis shows a 10,000-fold increase in transfer frequency following induction by the sex pheromone cAD1. Mutagenesis of the plasmid with transposon Tn917 was undertaken to determine the region(s) of pAD1 required for the mating response. The relevant genetic material was found to be distributed over a 31.2-kilobase contiguous region of the plasmid. Although insertions in two previously identified regions (traA and traB) exhibited increased transfer frequencies, insertions in five new regions (D, E, F, G, and H) decreased the ability of pAD1 to transfer. Insertions in region H allowed the cells to form visible mating aggregates, but the plasmid transfer frequency was decreased to levels below detection during a 1-h broth mating. Mutants with mutations in region G were able to form aggregates; however, insertions in regions D, E, and F prevented aggregate formation. Insertions in region C decreased the sensitivity of the cell to exogenous cAD1 and exhibited increased activity of the pheromone inhibitor iAD1. Surface protein profiles produced by a number of these mutants were examined, and in some cases were found to be different from those of the wild type. A map showing the various regions is presented, and related aspects of the regulation of the pAD1 mating response are discussed. PMID- 3038842 TI - Identification of endogenous inducers of the mal regulon in Escherichia coli. AB - The expression of the maltose regulon in Escherichia coli is induced when maltose or maltodextrins are present in the growth medium. Mutations in malK, which codes for a component of the transport system, result in the elevated expression of the remaining mal genes. Uninduced expression in the wild type, as well as elevated expression in malK mutants, is strongly repressed at high osmolarity. In the absence of malQ-encoded amylomaltase, expression remains high at high osmolarity. We found that uninduced expression in the wild type and elevated expression in malK mutants were paralleled by the appearance of two types of endogenous carbohydrates. One, produced primarily at high osmolarity, was identified as comprising maltodextrins that are derived from glycogen or glycogen-synthesizing enzymes. The other, produced primarily at low osmolarity, consisted of an oligosaccharide that was not derived from glycogen. We isolated a mutant that no longer synthesized this oligosaccharide. The gene carrying this mutation, termed malI, was mapped at min 36 on the E. coli linkage map. A Tn10 insertion in malI also resulted in the loss of constitutivity at low osmolarity and delayed the induction of the maltose regulon by exogenous inducers. PMID- 3038843 TI - Identification of uhp polypeptides and evidence for their role in exogenous induction of the sugar phosphate transport system of Escherichia coli K-12. AB - Cells of Escherichia coli possess a transport system that catalyzes the accumulation, in unaltered form, of a variety of sugar phosphates. Induction of the transport activity occurs in response to external glucose 6-phosphate and does not require detectable entry of this inducer. To define the genes that encode the Uhp transport system and those that mediate its exogenous induction, transposon insertions were isolated and mapped within a 6.5-kilobase HindIII BamHI fragment that carries the entire uhp region. The transposon insertions were transferred by homologous recombination onto the chromosome to test their effect on Uhp expression when all genes were present in single copy number. The complementation behavior of plasmids carrying the insertions or subcloned fragments of the region was compared with their polypeptide coding capacity in maxicells. These studies defined three uhp regulatory genes (uhpABC), all of which are necessary for expression of the uhpT gene, which encodes the transporter. The products of uhpB and uhpC are not required when uhpA is present on a multicopy plasmid. The four genes, uhpA, uhpB, uhpC, and uhpT, are transcribed in the same direction, and their products have apparent molecular weights of 25,000, 48,000, 20,000, and 38,000, respectively. The UhpB and UhpT polypeptides are associated with the membrane fraction. These results led to a model of regulation in which the UhpB and UhpC regulatory proteins prevent the ability of UhpA to activate transcription of the uhpT gene under noninducing conditions. PMID- 3038844 TI - Complete nucleotide sequence of insertion element IS4351 from Bacteroides fragilis. AB - The nucleotide sequence and genetic analyses of one of the directly repeated sequences flanking the macrolide-lincosamide-streptogramin B drug resistance determinant, ermF, from the Bacteroides fragilis R plasmid, pBF4, suggested that this region is an insertion sequence (IS) element. This 1,155-base-pair element contained partially matched (20 of 25 base pairs) terminal-inverted repeats, overlapping, anti-parallel open reading frames, and nine promoterlike sequences, including three that were oriented outward. Analysis of this sequence revealed no significant nucleotide homology to 13 other known IS elements. Inasmuch as Southern blot hybridization analysis detected homologous sequences in chromosomal DNA and its G+C content (42 mol%) was similar to that of B. fragilis, the data suggested that this element is of Bacteroides origin. Transposition promoted by this element was demonstrated in recA E. coli. Recombinants were recovered by selecting for the activation of a promoterless chloramphenicol resistance gene on the plasmid pDH5110 and were characterized by restriction endonuclease mapping and Southern blot hybridization. We propose that this IS element be designated IS4351. PMID- 3038845 TI - Clustering of mutations blocking synthesis of xanthan gum by Xanthomonas campestris. AB - Mutations that block the synthesis of xanthan gum by Xanthomonas campestris B1459S-4L-II were isolated as nonmucoid colonies after treatment with ethyl methanesulfonate. Complete libraries of DNA fragments from wild-type X. campestris were cloned into Escherichia coli by using a broad-host-range cosmid vector and then transferred into each mutant strain by conjugal mating. Cloned fragments that restored xanthan gum synthesis (Xgs+; mucoidy) were compared according to restriction pattern, DNA sequence homology, and complementation of a subset of Xgs- mutations. Groups of clones that contained overlapping homologous DNA were found to complement specific Xgs- mutations. The results suggest clustering of the genetic loci involved in xanthan synthesis. The clustering occurred within three unlinked regions. Two forms of complementation were observed. In most instances, independently isolated cosmid clones that complemented a single mutation were found to be partially homologous. Less frequent was the second form of complementation, in which two cosmid clones that lacked any homologous sequences restored the mucoid phenotype to a single mutant. Finally, xanthan production was measured for wild-type X. campestris carrying multiple plasmid copies of the cloned xanthan genes. PMID- 3038846 TI - Synthesis of deoxyribomononucleotides in Mollicutes: dependence on deoxyribose-1 phosphate and PPi. AB - Cell extracts of Acholeplasma laidlawii B-PG9, Acholeplasma morum S2, Mycoplasma capricolum 14, and Mycoplasma gallisepticum S6 were examined for 37 cytoplasmic enzyme activities involved in the salvage and biosynthesis of purines. All of these organisms had adenine phosphoribosyltransferase activity (EC 2.4.2.7) and hypoxanthine phosphoribosyltransferase activity (EC 2.4.2.8). All of these organisms had purine-nucleoside phosphorylase activity (EC 2.4.2.1) in the synthetic direction using ribose-1-phosphate (R-1-P) or deoxyribose-1-phosphate (dR-1-P); this activity generated ribonucleosides or deoxyribonucleosides, respectively. The pyrimidine nucleobase uracil could also be ribosylated by using either R-1-P or dR-1-P as a donor. The synthesis of deoxyribonucleosides from nucleobases and dR-1-P has been reported from only one other procaryote, Escherichia coli (L. A. Mason and J. O. Lampen, J. Biol. Chem. 193:539-547, 1951). The reverse of this phosphorylase reaction is more widely known, and we found such activity in all mollicutes studied. Some Acholeplasma species but not the Mycoplasma species can phosphorylate deoxyribonucleosides to deoxyribomononucleotides by a PPi-dependent deoxyribonucleoside kinase activity, which was first reported in this group for the ribose analogs (V. V. Tryon and J. D. Pollack, Int. J. Syst. Bacteriol. 35:497-501, 1985). This is the first report of PPi-dependent purine deoxyribonucleoside kinase activity. An ATP-dependent purine deoxyribonucleoside kinase activity is known only in salmon milt extracts (H. L. A. Tarr, Can. J. Biochem. 42:1535-1545, 1964). Deoxyribomononucleotidase activity was also found in cytoplasmic extracts of these mollicutes. This is the first report of deoxyribomononucleotidase activity. PMID- 3038847 TI - Cloning of the gene for phosphoribulokinase activity from Rhodobacter sphaeroides and its expression in Escherichia coli. AB - A 3.4-kilobase EcoRI restriction endonuclease fragment has been cloned from the facultatively photoheterotrophic bacterium Rhodobacter sphaeroides and shown to contain the structural gene (prkA) for phosphoribulokinase (PRK) activity. The PRK activity was characterized in Escherichia coli, and the product of the reaction was identified. The prkA gene was localized to a 1,565-base-pair EcoRI PstI restriction endonuclease fragment and gave rise to a 33-kilodalton polypeptide both in vivo and in vitro. The gene product produced in E. coli was shown to be identical to the gene product produced in R. sphaeroides. The amino acid sequence for the amino-terminal region deduced from the DNA sequence confirmed that derived for partially purified PRK derived from both E. coli and R. sphaeroides. In addition, the 3.4-kilobase EcoRI restriction endonuclease fragment coded for a 37-kilodalton polypeptide of unknown function, and preliminary evidence indicates that this DNA fragment is linked to genes coding for other activities significant in photosynthetic carbon assimilation. The genetic organization and proposed operon structure of this DNA fragment are discussed. PMID- 3038848 TI - Organization of phosphoribulokinase and ribulose bisphosphate carboxylase/oxygenase genes in Rhodopseudomonas (Rhodobacter) sphaeroides. AB - A heterologous phosphoribulokinase (PRK) gene probe was used to analyze two recombinant plasmids isolated from a Rhodopseudomonas (Rhodobacter) sphaeroides gene library. These plasmids were previously shown to carry the genes for form I and form II ribulose 1,5-bisphosphate carboxylase/oxygenase (RuBPC/O). Southern blot hybridization analysis indicated that there were two PRK genes linked to the RuBPC/O coding sequences. Restriction mapping showed the arrangement of the duplicate sets of PRK and RuBPC/O to be distinct. Subcloning of the hybridizing PRK sequences downstream of the lac promoter of pUC8 allowed expression of the two PRK enzymes in Escherichia coli. Analysis of the purified proteins by sodium dodecyl sulfate-slab gel electrophoresis revealed polypeptides with molecular weights of 32,000 and 34,000 corresponding to the form I and form II PRKs, respectively. Preliminary experiments on sensitivity to NADH regulation suggested that the two PRK enzymes differ in catalytic properties. PMID- 3038849 TI - Complementation of a trpE deletion in Escherichia coli by Spirochaeta aurantia DNA encoding anthranilate synthetase component I activity. AB - A 2.7-kilobase Sau3A fragment of Spirochaeta aurantia DNA cloned in pBR322 complemented a trpE deletion in Escherichia coli. Deletion analysis and Tn5 mutagenesis of the resulting plasmid pBG100 defined a 2-kilobase-pair region that was required for both the complementation and the synthesis of 59,000- and 47,000 molecular-weight polypeptides (59K and 47K polypeptides) in maxicells. Both the 59K and the 47K polypeptides appear to be encoded by a single gene. A maxicell analysis of pBG100::Tn5 mutants suggests that the 47K polypeptide is not sufficient for the trpE complementation. In vitro and in vivo anthranilate synthetase (AS) assays indicate that the complementing activity encoded by pBG100 was functionally analogous to the AS component I of E. coli in that it utilized NH3 but not glutamine as the amino donor. pBG100 did not encode a glutamine amidotransferase activity, although the AS component I it encoded was capable of interacting with E. coli AS component II to catalyze the glutamine-requiring reaction. Expression appeared to depend on a promoter in the cloned S. aurantia DNA. PMID- 3038850 TI - Cloning of the gene for myxobacterial hemagglutinin and isolation and analysis of structural gene mutations. AB - Myxobacterial hemagglutinin (MBHA) is a major developmentally induced protein that accumulates during the period of cellular aggregation in the bacterium Myxococcus xanthus. It has been shown that this lectin is targeted to the cell surface and periplasmic space of developmental cells, suggesting that it may play a role in cell-cell recognition or agglutination. We have cloned the structural gene for MBHA by using synthetic deoxyoligonucleotides containing sequences deduced from the amino acid sequence of MBHA and have used the cloned gene to construct strains of M. xanthus that cannot synthesize MBHA. We found that although the MBHA-deficient strains are delayed in their developmental time course, they are otherwise able to aggregate and sporulate normally. Our results suggest that MBHA may function to increase the efficiency of fruiting-body formation but is not a critical component of cellular aggregation. PMID- 3038851 TI - Heat shock and hydrogen peroxide responses of Escherichia coli are not changed by dinucleoside tetraphosphate hydrolase overproduction. AB - In Escherichia coli strains overproducing dinucleoside tetraphosphate hydrolase, the accumulation of dinucleoside tetraphosphates (AppppN, with N = A, C, G, or U) during heat shock or H2O2 treatment was reduced about 10-fold as compared with a control strain. This accumulation neither modified the pattern of the proteins induced by a temperature shift or H2O2 nor reduced the protection against oxidative damage induced by moderate H2O2 levels. PMID- 3038853 TI - Mesoridazine and thioridazine. PMID- 3038852 TI - Conjugal transfer of transposon Tn916 from Streptococcus faecalis to Mycoplasma hominis. AB - Transposon Tn916 was transferred from Streptococcus faecalis to Mycoplasma hominis by a mating process resembling conjugation with a frequency of 10(-6) to 10(-7). Tn916 was inserted into the mycoplasmal chromosome in single and multiple copies. PMID- 3038854 TI - Gene structure of human cytochrome P-450(SCC), cholesterol desmolase. AB - Four independent clones containing a part of the P-450(SCC), cholesterol desmolase, gene were isolated from human genomic libraries using bovine P 450(SCC) cDNA as a probe. These clones covered the entire P-450(SCC) gene except for a part of the 1st intron. The gene is at least 20 kb long and is split into 9 exons by 8 introns. The sequence analysis revealed that the nine separated exons code for a primary structure consisting of 521 amino acids which shows 72% homology with that of bovine P-450(SCC). A CATT sequence and a TATAAT sequence, which are possibly a "CAT" box, and a "TATA" box, respectively, are present 129 and 91 bp upstream from the initiation codon. An unusual exon/intron junctional sequence that begins with GC was found in the 6th intron of the gene. A putative extension peptide consisting of 39 amino acids was found in the sequence of human P-450(SCC) by comparison with that of the bovine counterpart. Two conserved regions were found in the extension peptide of these two forms of P-450(SCC), suggesting a functional role of the portions in the mitochondrial localization and processing of P-450(SCC) precursor. The mature form of human P-450(SCC) has only one cysteine residue, which was located in the center of the HR2 region (Gotoh et al. (1983) J. Biochem. 97, 807-817). This observation established beyond doubt that the sole cysteine residue in the HR2 region is the 5th ligand to the heme. PMID- 3038855 TI - E-F hand structure-domain of calcium-activated neutral protease (CANP) can bind Ca2+ ions. AB - The cDNA fragments corresponding to the domains with four consecutive E-F hand structures in the large and small subunits of chicken and rabbit calcium activated neutral protease (CANP) were inserted into an expression vector (pUC8 or pUC18). The resulting plasmids were used to transform E. coli, and isopropyl-1 thio-beta-D-galactoside (IPTG)-inducible expression was performed. The resulting four kinds of E-F hand structure-domains (the chicken large subunit, rabbit high- and low-calcium-requiring large subunits, and rabbit small subunit) were purified and analyzed for their calcium-binding abilities and capacities by the microscale filter assay. Most of the E-F hand structures could bind calcium and 2 or 4 mol of Ca2+ ions bound to the four consecutive E-F hand structures. The calcium binding affinity of the E-F hand structures in the large subunit roughly corresponds to the calcium concentration required for its CANP activity. PMID- 3038856 TI - Significance of phosphorylation/dephosphorylation of 46K protein(s) in regulation of superoxide anion production in intact guinea pig polymorphonuclear leukocytes. AB - Superoxide anion (O2-) production stimulated by concanavalin A (Con A) in guinea pig polymorphonuclear leukocytes (PMNL) was suppressed by addition of methyl alpha-mannoside, a Con A inhibitor, and resumed upon readdition of Con A. The reversible change in the O2- production was assumed to reflect the change in NADPH oxidase activity measured for the 30,000 X g particulate fraction. The stimulation by Con A of the phosphorylation of 46K protein(s), as observed previously with several membrane-perturbing agents in parallel with an activation of NADPH oxidase in intact guinea pig PMNL (Okamura, N., et al. (1984) Arch. Biochem. Biophys. 228, 270-277), was also suppressed by methyl-alpha-mannoside and resumed upon readdition of Con A. Similar parallelism between the phosphorylation and NADPH oxidase activity was also observed in the case of stimulation by N-formyl-methionyl-leucyl-phenylalanine (FMLP) and phorbol 12 myristate 13-acetate (PMA), though both processes were reversible after the stimulation by FMLP but not reversible after that by PMA. Thus, such a parallelism observed in both intact PMNL and 30,000 X g particulate fraction indicates possible involvement of the protein phosphorylation in the regulation of the production of active oxygen metabolites in PMNL. PMID- 3038857 TI - A unique specificity of a calcium activated neutral protease indicated in histone hydrolysis. AB - Calf thymus histones were found to be susceptible to a calcium-activated neutral protease [CANP: EC 3.4.22.17] which required a high concentration of calcium ions for its activity (mCANP). The susceptibilities of histones were in the order of relative degradation rate: H2B, H2A, and H3. The major peptide fragments released by CANP from H2A, H2B, and H3 were isolated and the cleavage sites were determined. Examination of amino acid sequences and environmental features around the cleavage site as well as kinetic analysis of the degradation process led us to the following conclusions about the mode of substrate recognition of mCANP: 1) The cleavage sites in histones could not be interpreted in terms of the primary structure around them. Thus, it seems unlikely that the specificity of CANP solely depends on its recognition of any specific amino acid residues or sequences. 2) The susceptible bonds were never located in the midst of either a hydrophobic or hydrophilic alignment of amino acid residues but in the vicinity of the boundary between hydrophilic and hydrophobic clusters. 3) Once a peptide fragment was generated by the proteolytic degradation, no further cleavage occurred even if the peptide still contained a bond corresponding to what was susceptible to CANP in an intact histone. This observation was interpreted to mean that CANP may recognize a certain higher order structure of its substrates. PMID- 3038858 TI - Comparative studies of the biological activities of human tumor necrosis factor and its derivatives. AB - Biological activities of human tumor necrosis factor (TNF) and its derivatives were compared. In cytotoxicity assay with L929 cells, one derivative, designated as TNF(Asn), showed significantly lower activity than any other TNF examined. In binding assay, this derivative was also shown to have lower affinity for TNF receptors on L929 cells, suggesting that the cytotoxic activity of TNFs on L929 cells correlates with their affinity for receptors. We also found that the cytotoxic activity of TNF on A673 cells and its inhibitory effect on lipoprotein lipase were parallel with the cytotoxic activity on L929 cells, but the growth enhancing activity on FS-4 cells and the cytotoxic activity on endothelial cells were not. It was also shown that TNF(Asn) had lower affinity than any other TNF for receptors on these target cells tested. These results suggested that there might be at least two types of cellular responses to TNF; one might correlate with the receptor-binding affinity of TNFs and the other not. PMID- 3038859 TI - Complete amino acid sequence of cytochrome c551 from Erythrobacter species strain OCh 114. AB - The complete amino acid sequence of cytochrome c551 isolated from an aerobic photosynthetic bacterium, Erythrobacter sp. strain OCh 114, was determined. The cytochrome molecule was composed of a total of 119 amino acid residues and its molecular weight including heme was calculated to be 13,235. The sequence was (Sequence: see text). Its molecular weight indicates that this cytochrome is of the L-type. Sequence alignment with other bacterial cytochromes c shows that this cytochrome is similar to cytochromes c of Rhodobacter capsulatus, Rhodobacter sphaeroides, and Paracoccus denitrificans, which were grouped into the alpha-3 subcluster from the 16S rRNA sequence analysis. PMID- 3038860 TI - The kinetics of intact microsome glucose-6-phosphatase are sigmoid at physiologic glucose 6-phosphate concentrations. AB - The kinetics of rat liver glucose-6-phosphatase (D-glucose-6-phosphate phosphohydrolase, EC 3.1.3.9) were studied with intact and detergent-disrupted microsomes from normal and diabetic rats. Glucose-6-P concentrations employed (12 microM to 1.0 mM) spanned the physiologic range. With the enzyme of intact microsomes from both groups, plots of v versus [glucose-6-P] were sigmoid. Hanes plots (i.e. [glucose-6-P]/v versus [glucose-6-P]) were biphasic (concave upwards). A Hill coefficient of 1.45 was determined with substrate concentrations between 12 and 133 microM. Disruption of microsomal integrity abolished these departures from classic kinetic behavior, indicating that sigmoidicity may result from cooperative interaction of glucose-6-P with the glucose-6-phosphatase system at the substrate translocase specific for glucose-6-P. With the enzyme from normal rats the [glucose-6-P] at which the enzyme was maximally sensitive to variations in [glucose-6-P] (which we term "Smax"), determined from plots of dv/d [glucose-6-P] versus [glucose-6-P], was in the physiologic range. The Smax of 0.13 mM corresponded well with the normal steady-state hepatic [glucose-6-P] of 0.16 mM, consistent with glucose-6-phosphatase's function as a regulatory enzyme. With the diabetic enzyme, in contrast, values were 0.30 and 0.07 mM for the Smax and steady-state level, respectively. We suggest that the decreasing sensitivity of glucose-6-phosphatase activity to progressively diminishing glucose-6-P concentration, inherent in its sigmoid kinetics, constitutes a mechanism for the preservation of a residual pool of glucose-6-P for other hepatic metabolic functions in the presence of elevated concentrations of glucose-6-phosphatase such as in diabetes. PMID- 3038862 TI - (2')3',5'-Bisphosphate nucleotidase. AB - (2')3',5'-Bisphosphate nucleotidase has been prepared in electrophoretically homogeneous form from guinea pig liver. The enzyme catalyzes the hydrolysis of the 2'- or 3'-phosphate from the appropriate nucleoside 2',5'- and 3',5' bisphosphates and is active with 3'-phosphoadenosine 5'-phosphosulfate and with coenzyme A but not with ATP. The 40,000-dalton protein is a monomer that requires Mg2+ for activity. PMID- 3038861 TI - The use of a specific fluorescence probe to study the interaction of adrenodoxin with adrenodoxin reductase and cytochrome P-450scc. AB - The single free cysteine at residue 95 of bovine adrenodoxin was labeled with the fluorescent reagent N-iodoacetylamidoethyl-1-aminonaphthalene-5-sulfonate (1,5-I AEDANS). The modification had no effect on the interaction with adrenodoxin reductase or cytochrome P-450scc, suggesting that the AEDANS group at Cys-95 was not located at the binding site for these molecules. Addition of adrenodoxin reductase, cytochrome P-450scc, or cytochrome c to AEDANS-adrenodoxin was found to quench the fluorescence of the AEDANS in a manner consistent with the formation of 1:1 binary complexes. Forster energy transfer calculations indicated that the AEDANS label on adrenodoxin was 42 A from the heme group in cytochrome c, 36 A from the FAD group in adrenodoxin reductase, and 58 A from the heme group in cytochrome P-450scc in the respective binary complexes. These studies suggest that the FAD group in adrenodoxin reductase is located close to the binding domain for adrenodoxin but that the heme group in cytochrome P-450scc is deeply buried at least 26 A from the binding domain for adrenodoxin. Modification of all the lysines on adrenodoxin with maleic anhydride had no effect on the interaction with either adrenodoxin reductase or cytochrome P-450scc, suggesting that the lysines are not located at the binding site for either protein. Modification of all the arginine residues with p-hydroxyphenylglyoxal also had no effect on the interaction with adrenodoxin reductase or cytochrome P-450scc. These studies are consistent with the proposal that the binding sites on adrenodoxin for adrenodoxin reductase and cytochrome P-450scc overlap, and that adrenodoxin functions as a mobile electron carrier. PMID- 3038863 TI - The collagen substrate specificity of human neutrophil collagenase. AB - The substrate specificity of human neutrophil collagenase was examined using both monomeric and fibrillar collagens. The neutrophil enzyme cleaved types I, II, and III collagens, but failed to attack types IV or V. Against monomeric collagen substrates at 25 degrees C, the neutrophil enzyme displayed values for the Michaelis constant (Km) of 0.6-1.8 X 10(-6) M, essentially indistinguishable from the substrate affinities that characterize human fibroblast collagenase. Catalytic rates, however, varied considerably; type I collagen was cleaved with a specificity (kappa cat/Km) some 20-fold greater than type III. Type II collagen was degraded with intermediate selectivity, approximately equal to 25% of the type I rate, but 450% that of type III. This specificity contrasted markedly with that of human fibroblast collagenase, which cleaved human type III collagen 15 fold faster than type I and greater than 500-fold more rapidly than type II. Interestingly, the 20-fold selectivity for type I over type III exhibited by neutrophil collagenase against monomeric collagens was largely abolished following the reconstitution of these substrates into insoluble fibrils, falling to a value of just 1.5-fold. The distinctive and opposite preference by the human fibroblast enzyme for monomeric type III collagen over type I (15-fold) was similarly reduced to less than 2-fold upon substrate aggregation. The transition from native soluble collagen monomers into insoluble fibrils appeared to be handled by both the human neutrophil and fibroblast collagenases with similar facility on type I substrates. By comparison, however, the neutrophil enzyme degraded type III collagen fibrils faster than would have been predicted from solution rates, while the fibroblast enzyme cleaved such fibrils much slower than expected from solution values. In exploring this phenomenon further, solvent deuterium isotope effects were measured. The deuterium studies suggest that neutrophil collagenase, acting on type III fibrils (kappa H2O/kappa D2O = 5.0), is less sensitive to factors which govern the availability of water at the relatively hydrophobic site of peptide bond hydrolysis in the collagen molecule than is fibroblast collagenase (kappa H2O/kappa D2O = 15.0). PMID- 3038864 TI - Protein kinase C is involved in adrenergic stimulation of pineal cGMP accumulation. AB - The amounts of cAMP and cGMP in the rat pinealocyte are regulated by norepinephrine acting through synergistic dual receptor mechanisms involving alpha 1- and beta-adrenoceptors (Vanecek, J., Sugden, D., Weller, J.L., and Klein, D.C. (1985) Endocrinology 116, 2167-2173; Sugden, L., Sugden, D., and Klein, D.C. (1986) J. Biol. Chem. 261, 11608-11612). Based on the available evidence, it appears that Ca2+-phospholipid-dependent protein kinase is involved in the alpha 1-adrenergic potentiation of beta-adrenergic stimulation of cAMP, but not in the stimulation of cGMP (Sugden, D., Vanecek, J., Klein, D.C., Thomas, T.P., and Anderson, W.B. (1985) Nature 314, 359-361). In the present study the role of protein kinase C in the adrenergic stimulation of cGMP was reinvestigated, with the purpose of determining whether protein kinase C activators would potentiate the effects of beta-adrenergic agonists on cGMP if cells were also treated with agents known to elevate intracellular free Ca2+. The protein kinase C activator 4 beta-phorbol 12-myristate 13-acetate (PMA) markedly elevated the cGMP content of beta-adrenergically stimulated pinealocytes which had also been treated with 1 microM A23187, 15 mM K+, or 1 microM ouabain. The effects of A23187 were blocked by EGTA and those of K+ were blocked by nifedipine, establishing the involvement of Ca2+. The stimulatory effects of PMA on cGMP accumulation were mimicked by other protein kinase C activators. PMA also stimulated cGMP accumulation in cells treated with cholera toxin (1 microgram/ml) and A23187 (1 microM), but not in cells treated only with cholera toxin. These results suggest that protein kinase C, which is activated in the pinealocyte by the alpha-adrenergic agonist phenylephrine, is probably involved in the adrenergic regulation of cGMP accumulation at a step distal to receptor activation. PMID- 3038865 TI - Nonselective inhibition of neutrophil functions by sphinganine. AB - Sphinganine has been proposed to be a specific inhibitor of protein kinase C. In the present study we have evaluated whether sphinganine is a convenient tool to probe for the role of protein kinase C in neutrophil function. Human neutrophils were loaded with the fluorescent probe quin2 and then tested in parallel for cytosolic free Ca2+, [Ca2+]i, membrane potential changes, O2- production, and exocytosis of primary granules (containing beta-glucuronidase) in response to various stimuli. In addition to inhibiting O2- production and exocytosis in a dose-dependent manner, sphinganine also blocked formyl-methionyl-leucyl phenylalanine-induced [Ca2+]i, transients. Furthermore, sphinganine inhibited exocytosis elicited by the calcium ionophore ionomycin. Although sphinganine blocked O2- production due to phorbol 12-myristate 13-acetate, the most striking finding was that the drug rendered the cells leaky. Thus, at similar concentrations as those inhibiting cellular functions, sphinganine was shown to lead to cell permeabilization, as assessed by release of quin2 and cytoplasmic markers into the extracellular medium, and changes in plasma membrane potential. We conclude, therefore, that sphinganine does not appear to be a suitable compound for the evaluation of the involvement of protein kinase C in neutrophil activation. PMID- 3038866 TI - Reconstitution of monomeric cytochrome c oxidase into phospholipid vesicles yields functionally interacting cytochrome aa3 units. AB - When the carbon monoxide complex of fully reduced cytochrome c oxidase, reconstituted into liposomes, is mixed with oxygen-containing buffer, complex kinetic progress curves are observed. This pattern is seen irrespective of whether the oxidase used in reconstitution is the dimeric or monomeric (subunit III-depleted) enzyme. These findings are interpreted in the light of similar experiments on the detergent-solubilized enzyme reported by Gibson and Greenwood (Gibson, Q.H., and Greenwood, C. (1963) Biochem. J. 86, 541-554) and confirmed by ourselves. We conclude that reconstitution of monomeric (subunit III-less) enzyme yields, preferentially, vesicles containing more than one functional unit, possibly associated as dimers. This result is of significance to our understanding of the relationships between aggregation state and proton pumping capacity of cytochrome oxidase. PMID- 3038867 TI - The role of sulfur-containing amino acids in superoxide production and modification of low density lipoprotein by arterial smooth muscle cells. AB - Extracellular superoxide (O2-.) was detected in cultures of monkey arterial smooth muscle cells as measured by the superoxide dismutase-inhibitable reduction of cytochrome c and acetylated cytochrome c. Reduction of cytochrome c by these cells required L-cystine in the incubation medium. A variety of other sulfur containing amino acids, including D-cystine, L-cystathionine, L-methionine, and djenkolic acid did not support O2-. generation when present at concentrations equimolar to L-cystine. At millimolar concentrations, the chelators EDTA and diethylene triamine penta-acetic acid inhibited O2-. production by smooth muscle cells. This effect was maximal when the chelator was present at the same concentration as the sum of the Ca2+ and Mg2+ in the medium, suggesting a role for these cations in O2-. generation by cells. Modification of low density lipoprotein (LDL) by arterial smooth muscle cells, as assessed by changes in lipid peroxide content, mobility on agarose gel electrophoresis, and apoprotein B fragmentation, was also L-cystine-dependent. LDL modification also required micromolar concentrations of the transition metal ion Cu(II) or Fe(III) and was inhibited by superoxide dismutase. LDL modified by smooth muscle cells in the presence of L-cystine and Cu(II) was taken up and degraded less well than native LDL by human skin fibroblasts, suggesting that recognition by the LDL receptor was lost. In contrast, LDL modified by smooth muscle cells was taken up and degraded to a greater degree than native LDL by mouse peritoneal macrophages, consistent with recognition by the scavenger receptor. These results indicate that monkey arterial smooth muscle cells produce O2-. and modify LDL by an L cystine-dependent process. This may involve reduction of cystine to a thiol, possibly cysteine or a cysteine-containing peptide such as glutathione. Sulfur containing amino acids may play a role in atherogenesis by supporting cell mediated generation of reactive oxygen species and modification of lipoprotein to a form recognized by the scavenger receptor. PMID- 3038868 TI - Phosphorylation in vivo of yeast (Saccharomyces cerevisiae) fructose-1,6 bisphosphatase at the cyclic AMP-dependent site. AB - In vivo labeled fructose-1,6-bisphosphatase was immunopurified from yeast (Saccharomyces cerevisiae) cells that had been incubated in the presence of [32P] orthophosphate. Tryptic peptides from labeled enzyme were mapped by high performance liquid chromatography. Most of the radioactivity was found to be associated with the peptide Arg9 through Arg24, the same peptide which had been previously shown to be phosphorylated in vitro by cAMP-dependent protein kinase (Rittenhouse, J., Harrsch, P. B., Kim, J. N., and Marcus, F. (1986) J. Biol. Chem. 261, 3939-3943). The amino acid sequence analysis suggests that phosphorylation occurs at the same site, Ser11. We have also determined the extent of phosphorylation at Ser11 of fructose-1,6-bisphosphatase in yeast cultures growing under various nutritional conditions by measuring the relative amounts of phospho- and corresponding dephosphopeptides in tryptic digests. Significant levels of phosphorylation of the enzyme were found in yeast cultures grown under gluconeogenic conditions that varied from 0.15 to 0.50 mol of phosphate per mol of enzyme subunit. However, phosphate incorporation rapidly increased to greater than 0.8 mol after addition of glucose to these cultures. An alternative technique, based solely on enzyme activity measurements, was also developed to estimate the extent of fructose-1,6-bisphosphatase phosphorylation in yeast cultures. The results obtained with this technique agreed with those obtained by high performance liquid chromatography of tryptic peptides. PMID- 3038869 TI - The gene and the primary structure of ornithine decarboxylase from Saccharomyces cerevisiae. AB - The nucleotide sequence was determined for a 3-kilobase genomic fragment containing the ornithine decarboxylase gene of Saccharomyces cerevisiae. The fragment contained two open reading frames. Gene disruption localized the ornithine decarboxylase gene to a 1398-nucleotide open reading frame. Transcription of the yeast gene initiated at several sites 171 to 211 nucleotides 5' of the translational start site. The 3' end of the transcript extended approximately 300 nucleotides beyond the end of the ornithine decarboxylase coding region and contained two copies of the yeast ARS core sequence. Translation of the ornithine decarboxylase gene appeared to initiate at the first AUG condon of the open reading frame based upon translational fusions with the Escherichia coli beta-galactosidase gene. Since no introns were apparent, the 1398-nucleotide open reading frame was predicted to encode a 466-amino acid protein with a calculated Mr = 52,369. The deduced protein differed significantly in size from previous reports on yeast ornithine decarboxylase, but was very similar in size to mammalian ornithine decarboxylase. When the predicted amino acid sequence of yeast ornithine decarboxylase was compared with that of the mouse enzyme, alignment of the sequences revealed that 40% of the amino acid residues were identical. Chou-Fasman predictions of the secondary structure of the two enzymes indicated that secondary structure was also highly conserved. PMID- 3038870 TI - Development of vasoactive intestinal peptide-responsive adenylate cyclase during enterocytic differentiation of Caco-2 cells in culture. Evidence for an increased receptor level. AB - The purpose of this work was to study vasoactive intestinal peptide (VIP) receptors and the adenylate cyclase response to VIP upon enterocytic differentiation of the human colon adenocarcinoma cell line Caco-2 in culture. The VIP-stimulated enzyme activity is very low, e.g. 20% above basal activity in undifferentiated cells (day 5) and is enhanced markedly at confluency reaching a maximum, e.g. 270%, above basal activity in fully differentiated cells (day 30). VIP potency is also slightly enhanced, the EC50 of VIP ranging from 0.31 nM at day 5 to 0.07 nM at day 30. Modifications of the adenylate cyclase system are not responsible for the development of VIP response. Indeed, forskolin-stimulated adenylate cyclase activity is unchanged during differentiation supporting no alteration of the enzyme catalytic subunit. The same holds true for NaF and guanosine 5'-(beta, gamma-imido)trisphosphate, indicating a constant activity of the guanine nucleotide regulatory unit which mediates hormonal stimulation of adenylate cyclase (Ns). This is further supported by the similar extent of cholera toxin-catalyzed [32P]ADP-ribosylation of the Ns protein that is observed during differentiation. In sharp contrast, a dramatic increase of VIP receptor concentration is observed ranging from 32 fmol/mg of protein at day 5 to 414 fmol/mg of protein at day 30. This is confirmed by affinity cross-linking experiments showing an increased specific incorporation of 125I-VIP in a major 66,000-dalton component during differentiation. A slight increase in receptor affinity is also observed during differentiation with Kd ranging from 0.39 nM at day 5 to 0.08 nM at day 30. These data indicate that one population of VIP receptors accumulates during Caco-2 cell differentiation, representing the crucial event in the development of adenylate cyclase response to the peptide. PMID- 3038871 TI - Leukotriene A4 hydrolase in the human lung. Inactivation of the enzyme with leukotriene A4 isomers. AB - Leukotriene A4 hydrolase from the human lung was purified to apparent homogeneity. The molecular weight (68,000-71,000), the amino acid composition, and the N-terminal amino acid sequence were similar to those of the human neutrophil enzyme but different from those of human erythrocyte enzyme. The lung enzyme was inactivated by its substrate, leukotriene A4. To elucidate the substrate and the inactivator specificity of this enzyme, we synthesized various geometric and positional isomers of leukotriene A4. 14,15-Leukotriene A4, leukotriene A4 methyl ester, and geometric isomers of leukotriene A4 could not serve as substrates, but they inactivated the enzyme. On the other hand, styrene oxide and (5S)-trans-5,6-oxide-8,10,14-cis-12-trans-eicosatetraenoic acid neither served as substrates nor inactivated the enzyme. These results indicate that whereas allylic epoxide structures of arachidonic acids are responsible for inactivation of the enzyme, the free carboxylic acid, 5,6-oxide, and the tetraene structure with the 7,9-trans-11,14-cis configuration are required as a substrate for leukotriene A4 hydrolase. PMID- 3038872 TI - Identification of insulin receptor tyrosine residues autophosphorylated in vitro. AB - To identify the autophosphorylation sites on the human insulin receptor (IR), partially purified human IR was incubated in vitro in the presence of insulin and manganese [gamma-32P]ATP so as to achieve near-maximal activation of the histone 2b kinase activity. Approximately 70% of all beta subunit [32P]phosphotyrosine resides on two tryptic peptide segments identified by microsequencing as IR precursor (Ullrich, A., Bell, J. R., Chen, E.-Y., Herrera, R., Petruzelli, L. M., Dull, T. J., Gray, A., Coussens, L., Liao, Y.-C., Tsubokawa, M., Mason, A., Seeburg, P. H., Grunfeld, C., Rosen, O. M., and Ramachandran, J. (1985) Nature 313, 756-761) 1144-1152 (tyrosine at 1146, 1150, 1151, designated peptide 5) and 1315-1329 (tyrosine at 1316, 1322, designated peptide 8), which were recovered in approximately equal amounts. Half of the remaining unidentified [32P]phosphotyrosine residues reside on another tryptic peptide of Mr 4000-5000. Assignment of [32P]phosphotyrosine to specific residues required subdigestion and Edman degradation of 32P peptides covalently coupled to solid supports. Peptide 5 was recovered in triple and double phosphorylated forms in a molar ratio of about 2:1. Tyr-1146 contained 32P in both forms of peptide 5; in the double phosphorylated form, phenylthiohydantoin-[32P]phosphotyrosine was recovered at both Tyr-1150 and Tyr-1151, in a ratio of about 1:2. Thus, the double phosphorylated peptide 5 is presumably a mixture of Tyr-P-1146/1150 and Tyr-P 1146/1151, predominantly the latter. Peptide 8 was recovered only as the double phosphorylated form. We conclude that autophosphorylation of human IR in vitro leads to the phosphorylation of at least 6 of the 13 tyrosine residues on the beta subunit intracellular extension. Five of these tyrosines are clustered in two domains; one domain is in the structurally unique C-terminal tail and contains Tyr-1316 and -1322 which are both phosphorylated. The second domain is located in the segment of the tyrosine kinase region homologous to the major in vitro autophosphorylation site of pp60 v-src and contains Tyr-1146, which is fully phosphorylated, and Tyr-1150 and -1151; although the majority of IR beta subunits exhibit phosphorylation of both tyrosine 1150 and 1151, up to 20-25% of Tyr-1150 remains unphosphorylated at complete kinase activation. PMID- 3038873 TI - Functional heterogeneity of atrial natriuretic factor receptor in bovine adrenal zona glomerulosa is explained by an amiloride-sensitive high affinity molecular complex. AB - The effects of amiloride on the molecular characteristics of the atrial natriuretic factor (ANF) receptor from bovine adrenal zona glomerulosa were studied by computer modeling of competitive binding data, by affinity labeling experiments, and by steric exclusion high performance liquid chromatography of solubilized receptor. The order of potency of a series of truncated ANF analogs in competing for 125I-ANF binding to bovine adrenal zona glomerulosa membranes was the same as that obtained for inhibition of aldosterone secretion. Deletion of amino acids at the COOH-terminal end drastically reduced the affinities of the peptides. Computer analysis of competition curves revealed that all ANF analogs tested show similar binding characteristics: shallow competition curves, discrimination of varying proportions of high and low affinity binding states, and sensitivity to amiloride which increases the proportion of the high affinity binding component. These results from binding studies are suggestive of potential heterogeneity of ANF binding sites. In contrast, results from affinity cross linking experiments are consistent with the notion of a single receptor protein. Incubation of membranes with increasing concentrations of 125I-ANF-(99-126) up to 3 nM resulted in the labeling of a single band of Mr 130,000. The ability of ANF analogs to compete for the labeling of the Mr 130,000 band by 125I-ANF-(99-126) agreed well with their potency as inhibitors of 125I-ANF binding to intact membranes. Addition of amiloride caused a dose-dependent increase in the labeling of the Mr 130,000 band. A single Mr 130,000 band was also labeled in bovine aorta and LLC-PK1 cell membranes. In order to further investigate the molecular basis for the apparent heterogeneity of ANF binding we have prelabeled the membrane receptor with 125I-ANF-(99-126) prior to solubilization with octyl-beta-D glucoside and chromatography on a Superose 6 steric exclusion column. The elution profile of the prelabeled receptor consistently showed two peaks of radioactivity with mean Stokes radii of 70 and 50 A. When amiloride was added to the incubation medium, the elution profile consisted almost exclusively of the 70-A peak. Quantitative analysis of the chromatographic profiles revealed that amiloride increases by 2-3 times the area of the 70-A peak. We conclude that the 70-A form represents a ternary complex of the receptor with an amiloride-sensitive effector protein. PMID- 3038874 TI - In vitro assembly of a prepriming complex at the origin of the Escherichia coli chromosome. AB - During initiation of DNA replication of plasmids containing the origin of the Escherichia coli chromosome (oriC), the proteins dnaA, dnaB, and dnaC interact and assemble a complex at oriC. The complex is larger and more asymmetric than that formed by dnaA protein and embraces an extra 50 base pairs at the left side of the minimal oriC sequence. Both dnaA and dnaB proteins have been identified in the complex by electron microscopy and antibody binding; dnaC protein was not detected. HU protein, which stimulates the activity of the initiation reaction, was often present. Entry of dnaB protein required dnaA and dnaC proteins and a supercoiled template. Thus, a complex structure, involving multiple proteins and a large region of DNA, must be formed at the origin to prepare the template for priming and replication. PMID- 3038875 TI - DNA gyrase-catalyzed decatenation of multiply linked DNA dimers. AB - One possible intermediate during the terminal stages of the replication of a closed circular DNA is a catenated DNA dimer of the two completed daughter molecules. The two monomer DNA rings in these DNA dimers can be linked as many as 20-30 times. In Escherichia coli, DNA gyrase could act on these catenated dimers to eliminate the linkages between the daughter duplexes, yielding the final monomer product. In this report, this reaction has been studied biochemically. The in vitro pBR322 DNA replication system (Minden, J., and Marians, K. J. (1985) J. Biol. Chem. 260, 9316-9325) was used to manufacture large amounts of multiply linked catenated DNA dimers for use as a substrate for DNA gyrase-catalyzed decatenation. Studies presented here demonstrate that this decatenation reaction is more efficient with supercoiled as opposed to relaxed DNA dimers, proceeds in a distributive fashion, is inhibited by moderate amounts of salt (80 mM KCl), and is stimulated by the E. coli protein HU. PMID- 3038876 TI - Energy transfer measurements of fusion between Sendai virus and vesicles corrected for decreased absorption of acceptor probe. AB - The fusion of Sendai virus at pH 4-7 with artificial lipid vesicles composed of phosphatidylserine or phosphatidylcholine was quantified by measuring fluorescence energy transfer from N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) phosphatidylethanolamine to N-(lissamine-rhodamine-B-sulfonyl) phosphatidylethanolamine in the target membranes. About 60% of the phosphatidylserine vesicles and virus appeared to fuse at pH 4 and about 100% at pH 5. Fusion was much less under all other conditions. The apparent fusion at pH 4, however, was due to a decrease in absorption of the acceptor probe, instead of dilution of acceptor as a result of fusion of labeled vesicles with unlabeled virus. After correction for this fusion-independent effect of Sendai virus, the extent of fusion was only 4-20% at pH 4 but still 80-100% at pH 5. These findings paralleled the loss of hemagglutinating and hemolytic activities of the virus induced by incubation at pH 4 but not at pH 5. Vesicle-virus hybrids were observed with the electron microscope after incubation at pH 5 but not at pH 7. The assay of membrane fusion by fluorescence energy transfer can be misleading unless correction is made for changes in energy transfer due to fusion independent effects. PMID- 3038877 TI - An investigation of hydrogenase I and hydrogenase II from Clostridium pasteurianum by resonance Raman spectroscopy. Evidence for a [2Fe-2S] cluster in hydrogenase I. AB - Resonance Raman spectra are reported for hydrogenase I and II from Clostridium pasteurianum. These spectra show overlapping bands with contributions from [4Fe 4S] clusters, known to be present in these enzymes, and from novel FeS centers of hitherto undefined structure. For hydrogenase I there are strong bands at 288 and 394 cm-1, which are seen in [2Fe-2S] proteins and in no other FeS species so far examined. In contrast these bands do not appear for hydrogenase II, whose resonance Raman spectrum is dominated by [4Fe-4S] cluster modes. These results provide the first structural information on the hydrogenase I FeS center involved in H2 activation and demonstrate structural differences between hydrogenase I and hydrogenase II. PMID- 3038878 TI - Rat carbamyl-phosphate synthetase I gene. Promoter sequence and tissue-specific transcriptional regulation in vitro. AB - The region flanking the 5'-end of the rat gene encoding the cytoplasmic precursor of carbamyl-phosphate synthetase I, a mitochondrial matrix enzyme, has been cloned and partially characterized. S1 nuclease and primer extension analyses position the starts of transcription 138-140 nucleotides upstream of the translation initiation codon. Exon 1 contains this untranslated sequence and extends downstream to include the coding region for the pre-enzyme signal peptide (38 amino acids) plus 4 amino acids from the amino terminus of the mature protein. The 5'-flanking sequence contains typical promoter elements, including putative TATA and CAAT motifs at -21 and -82 nucleotides, respectively. In addition, several copies of consensus sequences corresponding to the H4TF-1 recognition element, GATTTC, together with the enhancer-like octamer, ATTTGCAT, are also present. Carbamyl-phosphate synthetase I is a cell-type specific enzyme, being expressed only in hepatocytes and epithelial cells of the intestinal mucosa. It is also synthesized at relatively high levels in the hepatoma cell line, Hep G2. Employing pCPS2.1, a minigene containing the promoter and part of exon 1, we show that nuclear extracts from Hep G2 support accurate carbamyl phosphate synthetase I gene transcription in vitro. No such activity was observed, however, in extracts from HeLa, a cell line which does not express carbamyl-phosphate synthetase I. PMID- 3038879 TI - Intrinsic DNA-dependent ATPase activity of reverse gyrase. AB - Reverse gyrase is a type I DNA topoisomerase that promotes positive supercoiling of closed-circular double-stranded DNA through an ATP-dependent reaction, and it was purified from an archaebacterium, Sulfolobus. When ATP is replaced by UTP, GTP, or CTP, this enzyme just relaxes the negatively supercoiled closed-circular double-stranded DNA. We found that reverse gyrase hydrolyzes ATP through a double stranded DNA-dependent reaction. The superhelicity of the DNA did not affect the ATPase activity. However, reverse gyrase does not hydrolyze UTP, GTP, or CTP. Therefore, any of the four nucleotide 5'-triphosphates acts as an effector for the topoisomerase activity of reverse gyrase, but only ATP supports the positive supercoiling of closed-circular double-stranded DNA, through the energy released on its hydrolysis. Single-stranded DNA was a much more potent cofactor for the ATPase activity of the enzyme than double-stranded DNA, and it acted as a potent inhibitor for the topoisomerase activity on double-stranded DNA. These results indicate that reverse gyrase has higher affinity to single-stranded DNA than to double-stranded DNA, which suggests a cellular function of the enzyme. PMID- 3038880 TI - Identification of guanine nucleotides bound to ras-encoded proteins in growing yeast cells. AB - We have analyzed the guanine nucleotides bound to mammalian ras and yeast RAS proteins overexpressed in [32P]orthophosphate-labeled cultures of exponentially growing Saccharomyces cerevisiae cells. Whereas S. cerevisiae RAS1 and RAS2 proteins were immunoprecipitated bound entirely to GDP, mammalian Harvey ras was isolated with GTP and GDP bound in near-equimolar proportions. In a strain overexpressing a RAS2 variant where the RAS unique C-terminal domain was deleted, both GTP and GDP were detected in a ratio of 3:97. Increased amounts of GTP (16 75% of total guanine nucleotide) were observed bound to all ras proteins containing mutations that inhibit GTP hydrolytic activity. Increasing proportions of GTP bound to the various ras proteins correlated with increasing biological potency to bypass cdc25 lethality in yeast. PMID- 3038881 TI - Myeloperoxidase precursors incorporate heme. AB - Myeloperoxidase of neutrophil granulocytes is synthesized as a larger molecular weight precursor, which is processed to yield mature polypeptides with molecular weights of 62,000 and 12,000. We have investigated the incorporation of heme into myeloperoxidase of the human promyelocytic HL-60 cell line labeled with 5 amino[14C]levulinic acid. Myeloperoxidase was isolated by immunoprecipitation followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and radiolabeled myeloperoxidase was visualized by fluorography. A 3-h pulse labeling with 5-amino[14C]levulinic acid resulted in labeling of the Mr 90,000 and Mr 82,000 precursor polypeptides. During subsequent chase of the label, conversion to mature radioactive heavy Mr 62,000 subunit was observed but no radioactivity was associated with the mature small Mr 12,000 subunit. Peptide mapping after proteolytic cleavage with V8 proteinase showed that 5-amino[14C]levulinic acid was associated with a single Mr 23,000 polypeptide while multiple radioactive fragments were visible after proteolytic cleavage of myeloperoxidase biosynthetically labeled with [14C]leucine. That 5-amino[14C]levulinic acid was specifically incorporated into heme of myeloperoxidase was also demonstrated by dissociation under reducing conditions which yielded 14C-labeled heme as indicated by reversed phase high pressure liquid chromatography. The ionophore monensin and the base chloroquine, which block processing of myeloperoxidase, did not affect the incorporation of 5-amino[14C]levulinic acid, further supporting the notion that the incorporation of heme is independent of final processing of the polypeptide. Our data establish that heme is incorporated into myeloperoxidase already at the level of the precursor and that processing yields a heme-containing heavy subunit and a heme-free small subunit. PMID- 3038882 TI - Clostridial pyruvate oxidoreductase and the pyruvate-oxidizing enzyme specific to nitrogen fixation in Klebsiella pneumoniae are similar enzymes. AB - The chemical characterization, EPR properties, and mechanism of pyruvate:flavodoxin (ferredoxin) oxidoreductase from Klebsiella pneumoniae and Clostridium thermoaceticum have been investigated. A simple, specific, and sensitive assay and an efficient purification (based on the high affinity of these enzymes for a dye attached to agarose) are reported. The observed iron content of 8 atoms/subunit is twice that reported by others, whereas the contents of lipoate and flavin are less than 0.1 mol/subunit, in agreement with previous reports. Spectroscopic evidence suggests that the iron is present in Fe4S4(2+,1+) clusters. Reduction of the enzyme requires the presence of CoA as well as 1.1 pyruvate/subunit, which is very nearly the theoretical amount required the reduce two Fe4S(2+,1+) clusters. In the absence of CoA, stoichiometric amounts of pyruvate are decarboxylated, but the Fe/S centers are not reduced. We conclude that the K. pneumoniae and C. thermoaceticum enzymes are adapted to rapid reduction of low potential 1-e- carriers, similar to the pyruvate oxidoreductase of Halobacterium (Kerscher, L., and Oesterhelt, D. (1977) FEBS Lett. 83, 197 201), but different in that an Fe/S center-radical pair is used in the latter enzyme in place of the pair of Fe4S4 centers we find. The K. pneumoniae and C. thermoaceticum oxidoreductases appear to be mechanistically closely related to the Clostridium acidiurici enzyme (Uyeda, K., and Rabinowitz, J. C. (1971) J. Biol. Chem. 246, 3111-3119), differing as a class from the lipoate-containing, pyridine nucleotide-reducing enzyme present in aerobes (Reed, L. J. (1974) Accts. Chem. Res. 2, 740-746). The function of the Klebsiella enzyme is to supply electrons to nitrogenase. This is accomplished in vitro with purified components via a nif-specific flavodoxin or other low potential 1-e- carriers such as viologen dyes or ferredoxins. The in vivo molar ratio of nitrogenase to the physiological reduction system, estimated from activity measurements of individual components in crude extracts, was 0.4:0.03:2:1 pyruvate oxidoreductase:flavodoxin:nitrogenase component II:nitrogenase component I. PMID- 3038883 TI - Regulation of protein kinase C by cyclic adenosine 3':5'-monophosphate and a tumor promoter in skeletal myoblasts. AB - The specific activity of protein kinase C in rat skeletal myoblasts decreased when they were exposed for very short periods to isoproterenol, forskolin, dibutyryl cyclic AMP (Bt2cAMP), or the phorbol ester, 12-O-tetradecanoylphorbol 13-acetate (TPA). In the presence of Bt2cAMP or forskolin only the cytosolic but not the membrane-bound kinase activity was found to decrease. Treatment with TPA, however, led to a decrease in the activity of the enzyme both in the cytosolic as well as the membrane fractions. The effects observed in vivo could be duplicated in crude extracts of myoblasts incubated with cAMP analogues or TPA. In the presence of ATP, protein kinase C activity decreased considerably in crude cytosolic fractions treated with the cAMP analogues, but a requirement for ATP was not evident for the decrease in activity brought about by TPA. For the cAMP analogues the decrease in protein kinase C was also prevented by incubation of the extracts with an inhibitor of cAMP-dependent protein kinase. The regulation of protein kinase C by Bt2cAMP (but not by TPA) was altered in Rous sarcoma virus transformed myoblasts. It is considered likely that a component affected by cAMP (probably a substrate for cAMP-dependent protein kinase) participates in the regulation of protein kinase C activity, and it is altered in unknown ways in transformed myoblasts. PMID- 3038884 TI - Phosphorylation of the acetylcholine receptor by protein kinase C and identification of the phosphorylation site within the receptor delta subunit. AB - Purified acetylcholine receptor is rapidly and specifically phosphorylated by partially purified protein kinase C, the Ca2+/phospholipid-dependent enzyme. The receptor delta subunit is the major target for phosphorylation and is phosphorylated on serine residues to a final stoichiometry of 0.4 mol of phosphate/mol of subunit. Phosphorylation is dose-dependent with a Km value of 0.2 microM. Proteolytic digestion of the delta subunit phosphorylated by either protein kinase C or the cAMP-dependent protein kinase yielded a similar pattern of phosphorylated fragments. The amino acids phosphorylated by either kinase co localized within a 15-kDa proteolytic fragment of the delta subunit. This fragment was visualized by immunoblotting with antibodies against a synthetic peptide corresponding to residues 354-367 of the receptor delta subunit. This sequence, which contains 3 consecutive serine residues, was recently shown to include the cAMP-dependent protein kinase phosphorylation site (Souroujon, M. C., Neumann, D., Pizzighella, S., Fridkin, M., and Fuchs, S. (1986) EMBO J. 5, 543 546). Concomitantly, the synthetic peptide 354-367 was specifically phosphorylated in a Ca2+- and phospholipid-dependent manner by protein kinase C. Furthermore, antibodies directed against this peptide inhibited phosphorylation of the intact receptor by protein kinase C. We thus conclude that both the cAMP dependent protein kinase and protein kinase C phosphorylation sites reside in very close proximity within the 3 adjacent serine residues at positions 360, 361, and 362 of the delta subunit of the acetylcholine receptor. PMID- 3038885 TI - Occlusion of 22Na+ and 86Rb+ in membrane-bound and soluble protomeric alpha beta units of Na,K-ATPase. AB - In this work, we examined occlusion of 22Na+ and 86Rb+ in membranous and detergent-solubilized Na,K-ATPase from outer renal medulla. Optimum conditions for occlusion of 22Na+ were provided by formation of the phosphorylated complex from the beta,gamma-bidentate complex of chromium (III) with ATP (CrATP). Release of occluded cations occurred at equally slow rates in soluble and membrane-bound Na,K-ATPase. Values of 22Na+ occlusion as high as 11 nmol/mg of protein were measured, corresponding to 1.8-2.7 mol of Na+/mol of phosphorylated Na,K-ATPase as determined by 32P incorporation from [gamma-32P]CrATP. Maximum capacity for phosphorylation from [gamma-32P]CrATP was 6 nmol/mg of protein and equal to capacities for binding of [48V]vanadate and [3H]ouabain. The stoichiometry for occlusion of Rb+ was close to 2 Rb+ ions/phosphorylation site. In an analytical ultracentrifuge, the soluble Na+- or Rb+-occluded complexes showed sedimentation velocities (S20,w = 6.8-7.4) consistent with monomeric alpha beta-units. The data show that soluble monomeric alpha beta-units of Na,K-ATPase can occlude Rb+ or Na+ with the same stoichiometry as the membrane-bound enzyme. The structural basis for occlusion of cations in Na,K-ATPase is suggested to be the formation of a cavity inside a monomeric alpha beta-unit constituting the minimum protein unit required for active Na,K-transport. PMID- 3038887 TI - Regulatory domains of erythrocyte ankyrin. AB - This report provides evidence for regulatory domains of erythrocyte ankyrin that modulate associations of this protein with the anion transporter and spectrin. Two domains have been identified that are located at opposite ends of the polypeptide chain. One domain (Mr = 20,000), which is released by calpain, is primarily involved in regulation of the association of ankyrin with the anion transporter. The Mr = 195,000 fragment remaining after calpain cleavage binds to ankyrin-depleted inside-out vesicles with a 8-fold reaction in affinity, although with a 2-fold increase in number of high affinity sites. Cleavage of ankyrin by calpain induces a reduction in the frictional ratio from 1.55 to 1.33 suggesting either that the calpain-sensitive domain is present as a tail extending from a globular domain, or that upon cleavage ankyrin undergoes a major change in conformation. The other proposed regulatory domain is missing in protein 2.2, a form of ankyrin present in human erythrocytes that has a molecular weight about 29,000 smaller than ankyrin. Protein 2.2 is distinct from the calpain fragment based on peptide maps and reaction with domain-specific antibodies. The activity of the domain deleted from protein 2.2 has been inferred by comparison of ankyrin and protein 2.2, with the assumption that differences between these proteins are due to the missing domain. Protein 2.2 is an activated form of ankyrin that has a 3-fold higher affinity for spectrin and binds to twice the number of high affinity anion transporter sites. These observations suggested that removal of terminal domains of ankyrin may have a physiological role in modulation of ankyrin activity. PMID- 3038886 TI - Heme spin states and peroxide-induced radical species in prostaglandin H synthase. AB - We have examined the optical, magnetic circular dichroism, and electron paramagnetic resonance (EPR) spectra of pure ovine prostaglandin H synthase in its resting (ferric) and ferrous states and after addition of hydrogen peroxide or 15-hydroperoxyeicosatetraenoic acid. In resting synthase, the distribution of heme between high- and low-spin forms was temperature-dependent: 20% of the heme was low-spin at room temperature whereas 50% was low-spin at 12 K. Two histidine residues were coordinated to the heme iron in the low-spin species. Anaerobic reduction of the synthase with dithionite produced a high-spin ferrous species that had no EPR signals. Upon reaction with the resting synthase, both hydroperoxides quickly generated intense (20-40% of the synthase heme) and complex EPR signals around g = 2 that were accompanied by corresponding decreases in the intensity of the signals from ferric heme at g = 3 and g = 6. The signal generated by HOOH had a doublet at g = 2.003, split by 22 G, superimposed on a broad component with a peak at g = 2.085 and a trough at g = 1.95. The lipid hydroperoxide generated a singlet at g = 2.003, with a linewidth of 25 G, superimposed on a broad background with a peak at g = 2.095 and a trough around g = 1.9. These EPR signals induced by hydroperoxide may reflect synthase heme in the ferryl state complexed with a free radical derived from hydroperoxide or fragments of hydroperoxide. PMID- 3038888 TI - Insulin receptor kinase following internalization in isolated rat adipocytes. AB - We have studied how insulin-mediated internalization of insulin receptors and insulin activation of the insulin receptor kinase might be inter-related. Isolated rat adipocytes were exposed to 0, 6, or 500 ng/ml insulin for 40 min at 37 degrees C. Subsequently, plasma membrane, low-density microsomal membrane and high-density microsomal membrane subcellular fractions were prepared. Measurement of insulin binding to insulin receptors isolated from the membrane fractions revealed that exposure of cells to insulin resulted in a loss of binding activity (13% at 6 ng/ml, 27% at 500 ng/ml insulin) from the plasma membranes which was completely accounted for by the appearance of receptors in the low-density and high-density microsomal membrane fractions, indicating that insulin had induced translocation of insulin receptors from the surface to the cell interior. Measurement of kinase activity of the isolated receptors revealed that exposure of intact cells to 500 ng/ml insulin resulted in as much as a 35-fold increase in the intrinsic kinase activity of receptors from subcellular fractions. The kinase activity per receptor was equal in all fractions at 3-4 min but by 20 min the activity of the internalized receptors fell approximately 40% to a steady state; plasma membrane receptors, on the other hand, remained fully active over time. This indicates that newly internalized receptors retain their kinase activity but undergo subsequent deactivation. Following exposure of cells to 6 ng/ml insulin, the degree of activation of the insulin receptor kinase was lower in the plasma membrane fraction (24% of the insulin effect at 500 ng/ml) than in the low density and high-density microsomal membrane fractions (54 and 77%, respectively, of the insulin effect at 500 ng/ml). These results suggest that receptors with an activated kinase are preferentially internalized. We conclude that exposure of adipocytes to insulin causes endocytosis of insulin receptors and activation of insulin receptor kinase, newly internalized receptors are fully active tyrosine kinases but are deactivated as they traverse the intracellular organelles represented by low-density and high-density microsomal membranes, and insulin receptor occupancy, possibly by stimulating phosphorylation and activating the insulin receptor kinase, is important for targeting insulin receptors for internalization. PMID- 3038889 TI - Binding and internalization of platelet-activating factor 1-O-alkyl-2-acetyl-sn glycero-3-phosphocholine in washed rabbit platelets. AB - The binding profile of 1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine (AGEPC, platelet-activating factor) to washed rabbit platelets was investigated through the use of structural analogs of AGEPC, e.g. U66985, which specifically suppressed AGEPC biological activities on rabbit platelets. This interaction of AGEPC with platelets could be divided into three different components termed A, B, and C. Component A was considered as one of high affinity (Kd = 0.5 X 10(-9) M) and with a low capacity (about 400 sites/platelet). The binding of AGEPC to component A was reversible and was blocked by the inhibitory analogs of AGEPC. This was considered to be the AGEPC receptor site(s). Component B was irreversible in nature and was presumed to be associated with internalization of AGEPC. The latter process was sensitive to the structural inhibitors. Component C was not affected by the inhibitors and probably represented a nonspecific binding to the lipid layer of the membrane. The binding profile of 1-O-alkyl-2-(lyso)-sn glycero-3-phosphocholine, a biologically inactive and noninhibitory analog of AGEPC, was observed to consist of a single component and was (also) unaffected by the inhibitors. Internalization of AGEPC into rabbit platelets was further examined by the bovine serum albumin extraction method, which was originally developed by Mohandas et al. (Mohandas, N., Wyatt, J., Mel, S. F., Rossi, M. E., and Shohet, S. B. (1982) J. Biol. Chem. 257, 6537-6543). AGEPC was instantly taken up by the cell and internalization into its membrane, where it remained and was not released into cytosol. The internalization of AGEPC was suppressed by pretreating the cells with AGEPC analogs. In platelets desensitized to AGEPC, no down-regulation of the receptor site(s) was observed. The internalization of AGEPC in the desensitized cells was clearly enhanced and this was obvious even in the presence of the AGEPC inhibitor(s). Even in the presence of the inhibitors, effective internalization of AGEPC was also evident in thrombin-treated cells. These results suggested that the internalization of AGEPC was irreversibly enhanced in the platelets which were activated by AGEPC itself as well as by thrombin. PMID- 3038890 TI - Nonmuscle and smooth muscle myosin light chain mRNAs are generated from a single gene by the tissue-specific alternative RNA splicing. AB - We have isolated two cDNA clones for myosin alkali light chain (MLC) mRNA from two respective cDNA libraries of chick gizzard and fibroblast cells by cross hybridization to the previously isolated cDNA of skeletal muscle MLC. Sequence analysis of the two cloned cDNAs revealed that both of them are homologous to but distinct from the cDNA sequence used as the probe so that they may be classified into members of the MLC family, that they are identical with each other in the 3' and 5' untranslated sequence as well as in the coding sequence with a notable exception of a 39-nucleotide insertion in the fibroblast cDNA, 26 nucleotides of which are used for encoding the C-terminal amino acid sequence, and, therefore, that they encode the identical 142-amino acid sequence with different C-terminals of nine amino acids, each specific for fibroblast and gizzard smooth muscle MLC. The position of the inserted block corresponds exactly to one of the exon-intron junctions in the other MLC genes whose structures have so far been elucidated. DNA blot analysis suggested that the two MLC mRNAs of gizzard (smooth muscle) and fibroblast cells (nonmuscle) are generated from a single gene, probably through alternative RNA splicing mechanisms. RNA blot analysis and S1 nuclease mapping analysis using RNA preparations from fibroblast and gizzard tissues showed that the fibroblast MLC mRNA is expressed predominantly in fibroblast cells, but not, or very scantily if at all, in the gizzard, whereas the reverse is true for the gizzard smooth muscle MLC mRNA. PMID- 3038891 TI - cDNA sequence and tissue distribution of the mRNA for bovine and murine p11, the S100-related light chain of the protein-tyrosine kinase substrate p36 (calpactin I). AB - We have isolated and sequenced cDNA clones of bovine and murine p11 mRNAs. The nonpolyadenylated mRNAs are predicted to be 614 and 600 nucleotides, respectively. The p11 mRNAs both contain a 291 nucleotide open reading frame, preceded by a 5'-untranslated region of 73 nucleotides in bovine p11 mRNA and of 68 nucleotides in murine p11 mRNA. The deduced bovine p11 amino acid sequence is identical to the previously published partial bovine and complete porcine p11 protein sequence except for an additional COOH-terminal lysine residue. The bovine and murine p11 proteins are 92% homologous, whereas at the nucleotide level the conservation is 89% in the coding region and 75% in the 3'-untranslated region. Southern analysis of murine genomic DNA detected a single p11 gene, less than 10 kilobase pairs in size, containing as many as three introns. The p11 gene has been assigned to mouse chromosome 3 by analysis of interspecific hybrid cell panels and recombinant inbred mouse strains. The p11 gene is closely linked to the Xmmv-65 endogenous leukemia virus env gene and the guanylate binding protein 1 gene. Northern analyses of RNAs from mouse tissues and cell lines indicated that p11 mRNA levels vary widely. They are very low in liver, heart, and testes, moderate in brain, spleen, and thymus, and high in kidney, intestine, and lung. Analysis of the same RNA samples for p36 mRNA levels showed that expression of p11 and p36 mRNAs is not always coordinated. Brain and the mouse embryonal carcinoma cell line F9 contain moderate to high levels of p11 mRNA with very low levels of p36 mRNA. Sequence homology between p11 and the S100 proteins, and the serum-induced 2A9 gene product, as well as possible functions of p11 are discussed. PMID- 3038892 TI - Identification of highly reactive cysteinyl and methionyl residues of rabbit muscle phosphofructokinase. AB - The reactivity of the 16 thiol groups of rabbit skeletal muscle phosphofructokinase has been studied extensively over the past 20 years. Several of these thiols show high reactivity with a variety of reagents, display differential reactivity in the presence of allosteric ligands and substrates, and appear to be important to function because their modification changes activity and regulatory properties. In the present study, the location in the primary structure of several highly reactive thiol groups has been established by reaction with [14C]iodoacetate. In the course of these studies, 2 methionyl residues that are located at or near proposed ligand-binding sites are readily carboxymethylated by iodoacetate. In addition to confirming the presence of the most reactive thiol group at sequence position 88, a thiol protected from reaction by the presence of fructose-6-P and cyclic AMP has been found at position 169. Cysteine 169 is close to a residue important to the binding of fructose-6-P in the homologous structure from Bacillus stearothermophilis phosphofructokinase. The modification of Cys-169 brings about extensive, but not total, loss of activity. Another cysteine, at position 232, was found to be highly reactive also. Substrate provided partial protection against carboxymethylation at this position. Carboxymethylation of enzyme restricted to methionines 74 and 173 brought about no changes in the total activity or in the ATP inhibition profile of the enzyme. This is significant since position 74 was projected on the basis of the homologous procaryotic structure to be important in the binding of nucleotide to the allosteric site. PMID- 3038893 TI - Purification and partial characterization of a calmodulin-stimulated nucleoside triphosphatase from pea nuclei. AB - A nucleoside triphosphatase/deoxynucleoside triphosphatase associated with the chromatin fraction from a highly purified preparation of pea nuclei has been isolated and characterized. The purified enzyme has a molecular weight of 47,000 as checked by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and it has an isoelectric point of 6.6. In the presence of divalent cations (Mg2+ = Mn2+ greater than Ca2+), this enzyme hydrolyzes nucleoside triphosphates or deoxynucleoside triphosphates. Hydrolysis is optimal at pH 7.5 and is significantly inhibited by relatively low concentrations of quercetin, but is not sensitive to vanadate, nitrate, or oligomycin. The enzyme has a rather broad nucleotide substrate specificity and has a Km for MgATP2- of 0.6 mM. The enzyme activity is stimulated over 3-fold by Ca2+ and calmodulin, and the stimulation is blocked by the Ca2+ chelator EGTA and by the calmodulin antagonists compound 48/80 and chlorpromazine. PMID- 3038894 TI - Cloning and sequence analysis of the mouse genomic locus encoding the largest subunit of RNA polymerase II. AB - The genomic locus (RPII215) encoding the largest subunit of mouse RNA polymerase II has been cloned by low stringency hybridization to a Drosophila RPII215 probe. The mouse gene consists of 28 exons which span 30 kilobases. Analysis of the nucleotide and predicted protein sequences indicates that the protein is comprised of two domains. There is a 1500 residue amino-terminal domain which contains seven regions strikingly similar to those in the beta' subunit of Escherichia coli RNA polymerase, and a carboxyl-terminal domain comprised of 52 repeats of a 7-amino-acid consensus sequence Tyr-Ser-Pro-Thr-Ser-Pro-Ser. Among the seven highly conserved regions are a strongly basic domain consistent with a DNA-binding site and a consensus sequence characteristic of a potential zinc binding domain. The 5' upstream region contains three tandem sequences similar to binding sites for the transcription factor SP1. Two of the introns in this gene splice at donor GC dinucleotides as opposed to previously described invariant GT sites. The identification of regions which are highly conserved as compared with bacterial and yeast RNA polymerase and other regions which are unique to the mouse protein suggests which domains of RNA polymerase large subunits are involved in aspects of transcription common to both procaryotes and eucaryotes and which are characteristic of transcription in higher organisms. PMID- 3038895 TI - Expression of hybrid (Na+ + K+)-ATPase molecules after transfection of mouse Ltk cells with DNA encoding the beta-subunit of an avian brain sodium pump. AB - A cDNA encoding the beta-subunit of the (Na+ + K+)-ATPase was cloned from a chicken brain cDNA library, and its nucleotide sequence was determined. High cross-species sequence homologies were found both in coding and noncoding regions. The cDNA was subcloned into a shuttle vector derived from pSV2CAT and was stably incorporated into mouse Ltk-cells. The avian beta-subunit was expressed on the cell surface (1-8 X 10(5) molecules/cell) complexed with alpha subunits of the murine (Na+ + K+)-ATPase. In the hybrid system there was rapid assembly of subunits, post-translational N-glycosylations of the beta-subunit at its three Asn-X-Ser (or Thr) positions, and modification of high mannose oligosaccharides to complex type. Avian beta-subunits expressed in the mouse cells had an apparent molecular weight of about 55,000 as compared with 47,000 in avian cells, due to post-translational modifications, presumably differences in complex oligosaccharides. Despite the high number of interspecies hybrid (Na+ + K+)-ATPase molecules, the cells had none of the high affinity ouabain binding sites (KD = 2 X 10(-7) M) characteristic of avian cells, consistent with the view that the ouabain binding site is located largely or exclusively on the alpha subunit and is not greatly affected by alpha-beta interaction. PMID- 3038896 TI - Analysis of the 5' end of the Drosophila muscle myosin heavy chain gene. Alternatively spliced transcripts initiate at a single site and intron locations are conserved compared to myosin genes of other organisms. AB - We have localized the transcription start site of the Drosophila melanogaster muscle myosin heavy chain (MHC) gene and find that all forms of the alternatively spliced MHC mRNA initiate at the same location. Therefore the alternative inclusion/exclusion of the 3' penultimate exon in transcripts from this gene (Bernstein, S.I., Hansen, C.J., Becker, K.D., Wassenberg, D.R., II, Roche, E.S., Donady, J.J., and Emerson, C. P., Jr. (1986) Mol. Cell. Biol. 6, 2511-2519; Rozek, C.E., and Davidson, N. (1986) Proc. Natl. Acad. Sci. U.S.A. 83, 2128-2134) does not result from the use of different 5' transcription initiation sites. This gene is the first invertebrate MHC gene shown to have TATA and CAAT box consensus sequences and a noncoding 5' exon, properties that are shared with some vertebrate and invertebrate contractile protein genes. The intron that splits the 5' noncoding region of the Drosophila MHC gene contains no major conserved elements relative to other Drosophila contractile protein genes. The introns within the coding region near the 5' end of the Drosophila MHC gene are located at the same sites as nematode and vertebrate MHC gene introns, indicating that these MHC genes are derived from a common ancestral sequence. The putative ATP binding domain encoded in the fourth exon of the Drosophila MHC gene is highly conserved relative to vertebrate, invertebrate, and non-muscle MHC genes suggesting that each of these myosins bind ATP by the same mechanism. Two divergent copies of the third exon are present within the 5' region of the Drosophila MHC gene, suggesting that alternative splicing produces MHC isoforms with different globular head regions. PMID- 3038897 TI - Identification and purification of a kidney membrane protein which specifically binds the amino-terminal domain of native parathyroid hormone. AB - Nitrocellulose blots of bovine kidney membrane proteins were prepared from denaturing polyacrylamide gels. Strips of the blots were incubated with parathyroid hormone (PTH), washed, and then incubated with antisera against the hormone. Exposure to horseradish peroxidase-linked second antibody led to staining of a 51-kDa protein. No staining was observed in blots not incubated with PTH. Fragments 35-84 and 19-84 of PTH reacted strongly with the antisera, but did not lead to staining of the 51-kDa protein on the blots. Staining was visible, but greatly reduced, when fragment 9-84 was used. Oxidation of the native hormone at positions 8 and 18 led to reductions in staining of the band which were quantitatively similar to the reductions in biological activity induced by such oxidations. These properties suggested that the 51-kDa protein recognizes the amino-terminal portions of PTH, which is the segment of the molecule required for its biological activities. Several micrograms of the 51-kDa protein were purified to homogeneity by selective extraction from the membranes with detergent and by elution from multiple two-dimensional gels. The purified protein retained its PTH-dependent staining and specificity. This protein may be a PTH receptor or a fragment of a PTH receptor from kidney. PMID- 3038898 TI - On the fidelity of DNA replication. Isolation of high fidelity DNA polymerase primase complexes by immunoaffinity chromatography. AB - Error rates for conventionally purified DNA polymerase-alpha from calf thymus, chicken, and human sources have been reported to be one in 10,000 to one in 40,000 nucleotides incorporated. Isolation of polymerase-alpha by immunoaffinity chromatography yields a multiprotein high molecular weight replication complex that contains an associated DNA primase (Wong, S. W., Paborsky, L. R., Fisher, P. A., Wang, T. S-F., and Korn, D. (1986) J. Biol. Chem. 261, 7958-7968). We have isolated DNA polymerase-primase complexes from calf thymus, from a human lymphoblast cell line (TK-6), and from Chinese hamster lung cells (V-79) using two different methods of immunoaffinity chromatography. These enzyme complexes are 12- to 20-fold more accurate than conventionally purified calf thymus DNA polymerase-alpha when assayed using the phi X174am3 fidelity assay; estimated error rates are one in 460,000 to one in 830,000 nucleotides incorporated when the enzyme complex is freshly isolated. The polymerase-primase complex from calf thymus exhibited no detectable 3'----5' exonuclease activity using a heteroduplex substrate containing a single 3'-terminal mismatched nucleotide. Upon prolonged storage at -70 degrees C, the error rate of the immunoaffinity-purified calf thymus DNA polymerase-primase complex increases to about one in 50,000 nucleotides incorporated, an error rate similar to that exhibited by conventional isolates of DNA polymerase-alpha. PMID- 3038899 TI - In vitro initiation of DNA replication in simian virus 40 chromosomes. AB - A soluble system has been developed that can initiate DNA replication de novo in simian virus 40 (SV40) chromatin isolated from virus-infected monkey cells as well as in circular plasmid DNA containing a functional SV40 origin of replication (ori). Initiation of DNA replication in SV40 chromatin required the soluble fraction from a high-salt nuclear extract of SV40-infected cells, a low salt cytosol fraction, polyethylene glycol, and a buffered salts solution containing all four standard deoxyribonucleoside triphosphates. Purified SV40 large tumor antigen (T-ag) partially substituted for the high-salt nucleosol, and monoclonal antibodies directed against SV40 T-ag inhibited DNA replication. Replication began at ori and proceeded bidirectionally to generate replicating DNA intermediates in which the parental strands remained covalently closed, as observed in vivo. Partial inhibition of DNA synthesis by aphidicolin resulted in accumulation of newly initiated replicating intermediates in this system, a phenomenon not observed under conditions that supported completion of replication only. However, conditions that were optimal for initiation of replication repressed conversion of late-replicating intermediates into circular DNA monomers. Most surprising was the observation that p-n-butylphenyl-dGTP, a potent and specific inhibitor of DNA polymerase-alpha, failed to inhibit replication of SV40 chromatin under conditions that completely inhibited replication of plasmid DNA containing the SV40 ori and either purified or endogenous DNA polymerase alpha activity. In contrast, all of these DNA synthesis activities were inhibited equally by aphidicolin. Therefore, DNA replication in mammalian cells is carried out either by DNA polymerase-alpha that bears a unique association with chromatin or by a different enzyme such as DNA polymerase-delta. PMID- 3038900 TI - Bacteriophage T4 DNA primase-helicase. Characterization of oligomer synthesis by T4 61 protein alone and in conjunction with T4 41 protein. AB - The bacteriophage T4 41 and 61 proteins function as a primase-helicase which in vitro both unwinds double-stranded DNA and synthesizes the pentaribonucleotides used to initiate DNA synthesis on the lagging strand. We demonstrate that 61 protein alone possesses a weak DNA template-dependent oligomer synthesizing activity, whose products differ in size and nucleotide specificity from those made by the 61 and 41 proteins together. We have previously shown that the 61 and 41 proteins make primarily ribonucleotide pentamers of the sequence pppApC(pN)3, although some pentamers beginning with G were also detected on phi X174 single stranded DNA. The pentamers pppApC(pN)3 have also been shown to initiate T4 DNA chains in vivo (Kurosawa, Y., and Okazaki, T. (1979) J. Mol. Biol. 135, 841-861). We now show that in contrast, the major products made by 61 protein alone on phi X174 DNA with [alpha-32P]CTP and the other three ribonucleoside triphosphates are not pentamers, but the dimers pppApC and pppGpC. In addition, minor amounts of products from 3 to approximately 45 nucleotides in length are also synthesized. Unlike the 61/41 protein reaction, 61 protein alone can substitute dATP or dGTP for ATP or GTP. Addition of 41 protein greatly stimulates oligomer synthesis, especially the synthesis of products made with ATP and CTP and products 5 nucleotides in length. Thus, both 61 and 41 proteins are needed to obtain efficient synthesis of the biologically relevant pentamers pppApC(pN)3. We demonstrate that the glucosylated hydroxymethylcytosines present in T4 DNA do not support the initiation of primer synthesis by the 61 protein on this template. With glycosylated hydroxymethyl T4 DNA, pppApC but not pppGpC oligomers are detected. If the T4 DNA is modified by hydroxymethylation but not glucosylation, pppApC and only a trace of pppGpC products are seen. In the accompanying paper (Nossal, N.G., and Hinton, D.M. (1987) J. Biol. Chem. 262, 10879-10885), we examine DNA synthesis primed by 61 protein in the absence of 41 protein. PMID- 3038901 TI - Bacteriophage T4 DNA primase-helicase. Characterization of the DNA synthesis primed by T4 61 protein in the absence of T4 41 protein. AB - The bacteriophage T4 61/41 protein primase-helicase is part of a seven T4 protein system needed for DNA synthesis in vitro. Although both 41 and 61 proteins are required for the synthesis and utilization of the normal pppApC(pN)3 pentanucleotide primer, we show in the accompanying paper (Hinton, D. M., and Nossal, N. G. (1987) J. Biol. Chem. 262, 10873-10878) that high concentrations of 61 protein alone carry out a limited, template-dependent oligonucleotide synthesis with the dimers pppApC and pppGpC as the major products labeled with [alpha-32P]CTP. At these high concentrations, 61 protein alone primes DNA synthesis by T4 DNA polymerase and the T4 genes 44/62 and 45 polymerase accessory proteins, or by Escherichia coli DNA polymerase I. The addition of T4 replication proteins other than 41 protein does not change the size distribution of oligonucleotides made by 61 protein. However, the primers used for DNA synthesis in the absence of 41 protein are not dimers, but rather trace quantities of longer oligonucleotides (5 to about 45 bases) which begin predominantly with pppGpC. These results show that 41 protein is required to prime with oligonucleotides beginning with pppApC and suggest that 41 protein, either alone or in conjunction with 61 protein, helps to stabilize the usual short pentamer primers on the template until they are elongated by the DNA polymerase. Moreover, since 61 protein by itself can only initiate DNA synthesis with primers beginning with pppGpC, but cannot make oligonucleotides starting with pppGpC on T4 DNA in which all the C is glucosylated and hydroxymethylated, both the T4 41 and 61 proteins are essential to prime DNA synthesis on their normal template. In our analysis of RNA-primed DNA, we demonstrate that although RNA primers at the 5' ends of DNA chains are relatively resistant to the 3' to 5' exonuclease of T4 DNA polymerase (Kurosawa, Y., and Okazaki, T. (1979) J. Mol. Biol. 135, 841-861), pppNpNpNpNpN oligomers are digested to a greater extent than the dephosphorylated pentamers NpNpNpNpN. PMID- 3038902 TI - Hydride transfer stereospecificity of rat liver aldehyde dehydrogenases. AB - The stereospecificity of hydride transfer to NAD+ by several forms of rat liver aldehyde dehydrogenase was determined by a nuclear magnetic resonance method. The forms included several mitochondrial and microsomal isozymes from normal liver, as well as isozymes from xenobiotic-treated and tumor cells. The proton added to NAD+ comes exclusively from the aldehyde substrate and in all cases was A (pro-R) stereospecific. PMID- 3038903 TI - Inhibition by amine bases or by sodium ions and protection by divalent cations in the hydrolysis of phosphoenzyme of (Na,K)-ATPase. AB - In the absence of K+, the hydrolysis of low-energy phosphoenzyme (E2P) of (Na,K) ATPase, but not Ca-ATPase of sarcoplasmic reticulum, was inhibited by Na+ or by amine compounds, such as Tris, imidazole, arginine, or lysine, with half-maximum inhibition concentrations of millimolar order at pH 7.4 and 0 degrees C. Histidine was slightly inhibitory. Divalent cations alone were also inhibitors. However, divalent cations, especially Ca2+ or Mn2+, were apparently activators in the presence of another inhibitor, Na+, or an amine compound, probably because the inhibitory action of the divalent cations was less powerful than that of Na+ or amines, which appear to compete for the same binding site on E2P. In most reported experiments on the hydrolysis of the phosphoenzyme, Tris or imidazole (as a buffer component) and Na+ (to obtain a sufficient amount of phosphoenzyme) have been present in the reaction mixture in amounts that are now seen to be inhibitory. Consequently, the reactivity of the phosphoenzyme with water would have been substantially underestimated. Under these inhibitory conditions, however, reactivity with K+ or ADP was little influenced. PMID- 3038904 TI - The reaction of cytochromes c and c2 with the Rhodospirillum rubrum reaction center involves the heme crevice domain. AB - In order to define the interaction domain on Rhodospirillum rubrum cytochrome c2 for the photosynthetic reaction center, positively charged lysine amino groups on cytochrome c2 were modified to form negatively charged carboxydinitrophenyl lysines. The reaction mixture was separated into six different fractions by ion exchange chromatography on carboxymethylcellulose and sulfopropyl-Sepharose. Peptide mapping studies indicated that fraction A consisted of a mixture of singly labeled derivatives modified at lysines 58, 81, and 109 on the back of cytochrome c2. Fractions C1, C2, C3, and C4 were found to be mixtures of singly labeled derivatives modified at lysines 9, 13, 75, 86, and 88 on the front of cytochrome c2 surrounding the heme crevice. The photooxidation of the carboxydinitrophenyl-cytochrome c2 derivatives by reaction centers purified from R. rubrum was measured following excitation with a laser pulse. The second-order rate constant of fraction A modified at backside lysines was found to be 2.3 X 10(7) M-1 s-1, nearly the same as that of native cytochrome c2, 2.6 X 10(7) M-1 s 1. However, the rate constants of fractions C1-C4 were found to be 6 to 12-fold smaller than that of native cytochrome c2. These results indicate that lysines surrounding the heme crevice of cytochrome c2 are involved in electrostatic interactions with carboxylate groups at the binding site of the reaction center. The reaction rates of horse heart cytochrome c derivatives modified at single lysine amino groups with trifluoroacetyl or trifluoromethylphenylcarbamoyl were also measured. Modification of lysines 8, 13, 25, 27, 72, 79, or 87 surrounding the heme crevice was found to significantly lower the rate of reaction, while modification of lysines in other regions had no effect. This indicates that the reaction of horse heart cytochrome c with the reaction center also involves the heme crevice domain. PMID- 3038905 TI - In vitro methylation of the 5'-flanking regions of the mouse beta-globin gene. AB - The enzymatic methylation of the 5'-flanking region of the mouse beta-globin (major) gene containing putative regulatory regions has been investigated. In vitro methylation of this 368-base pair regulatory DNA by a DNA methyltransferase obtained from mouse erythroleukemia cells yields an asymmetric methylation pattern. Of the 10 available CG pairs, only 5-6 are modified, leading to one hemimethylated site and two apparently fully methylated sites. Only CG pairs which are localized in a 29-base pair cluster are methylated. The data suggest that a CG cluster approximately 100 base pairs upstream from the CAP site may be the in vivo site of methylation in the 5'-regulator region of the mouse beta globin gene. PMID- 3038906 TI - Separate binding sites for nuclear factor 1 and a CCAAT DNA binding factor in the mouse alpha 2(I) collagen promoter. AB - A factor present in nuclear extracts of NIH-3T3 fibroblasts was identified which binds to a region between -315 and -295 with respect to the start of transcription in the mouse alpha 2(I) collagen promoter. This factor was assayed using three different DNA binding assays. Evidence that this factor is nuclear factor 1 or a related protein is based on competition experiments using fragments of another promoter that contains a nuclear factor 1 binding site as well as a fragment containing a consensus sequence for nuclear factor 1 binding sites. The nucleotide sequence of the binding site in the alpha 2(I) promoter. TCGN5GCCAA, presents similarities to the sequence published previously as a consensus recognition sequence for nuclear factor 1. Methylation interference experiments indicate that the factor interacts at least with two successive G residues in the major groove complementary to the two successive C residues in the GCCAA motif. This factor is different from a factor that binds to a CCAAT element between -84 and -80 in the mouse alpha 2(I) collagen promoter. This was shown by competition experiments using both crude NIH-3T3 fibroblast nuclear extracts and a partially purified CCAAT binding factor. These results suggest that in the alpha 2(I) collagen promoter nuclear factor 1 and a CCAAT binding factor bind to separate sites, despite analogies in their recognition sequences. PMID- 3038907 TI - Semiquinone anion radicals from addition of amino acids, peptides, and proteins to quinones derived from oxidation of catechols and catecholamines. An ESR spin stabilization study. AB - Either chemical or enzymatic oxidation of catechols or catecholamines in the presence of nucleophiles (amino acids, peptides, and proteins) leads to the production of ring-substituted o-semiquinones which have been detected by ESR spin stabilization techniques. In many cases, radicals have been completely characterized and structures assigned. Chemical considerations point to a mechanism involving addition of nucleophile to o-quinone, followed by oxidation of product to o-semiquinone. These results confirm that addition occurs in oxidizing polyhydroxy aromatic systems, probably via o-quinone, in a reaction considered to account for much of the toxicity found for catechols and catecholamines. PMID- 3038908 TI - Involvement of the 24-kDa cap-binding protein in regulation of protein synthesis in mitosis. AB - The rate of protein synthesis in metaphase-arrested cells is reduced as compared to interphase cells. The reduction occurs at the translation initiation step. Here, we show that, whereas poliovirus RNA translation is not affected by the mitotic translational block, the translation of vesicular stomatitis virus mRNAs is. In an attempt to elucidate the mechanism by which initiation of protein synthesis is reduced in mitotic cells, we found that the interaction of the mRNA 24-kDa cap-binding protein (CBP) with the mRNA 5' cap structure is reduced in mitotic cell extracts, consistent with their lower translational efficiency. Addition of cap-binding protein complex stimulated the translation of endogenous mRNA in extracts from mitotic but not interphase cells. In addition, we found that the 24-kDa CBP from mitotic cells was metabolically labeled with 32P to a lesser extent than the protein purified from interphase cells. These results are consistent with a hypothesis that the 24-kDa CBP is implicated in the inhibition of protein synthesis in metaphase-arrested cells. Possible mechanisms for this inhibition are offered. PMID- 3038909 TI - Opal suppressor phosphoserine tRNA gene and pseudogene are located on human chromosomes 19 and 22, respectively. AB - An opal suppressor phosphoserine tRNA gene and pseudogene have been isolated from a human DNA library and sequenced (O'Neill, V., Eden, F., Pratt, K., and Hatfield, D. (1985) J. Biol. Chem. 260, 2501-2508). Southern hybridization of human genomic DNA with an opal suppressor tRNA probe suggested that the gene and pseudogene are present in single copy. In this study, we have determined the chromosome location of the human gene and pseudogene by utilizing a 193-base pair fragment encoding the opal suppressor phosphoserine tRNA gene as probe to examine DNAs isolated from human-rodent somatic cell hybrids that have segregated human chromosomes. These studies show that the probe hybridized with two regions in the human genome; one is located on chromosome 19 and the second on chromosome 22. By comparing the restriction sites within these two regions to those previously determined for the human opal suppressor phosphoserine tRNA gene and pseudogene, we tentatively assigned the gene to chromosome 19 and the pseudogene to chromosome 22. These assignments were confirmed by utilizing a 350-base pair fragment which was isolated from the 5'-flanking region of the human gene as probe. This fragment hybridized only to chromosome 19, demonstrating unequivocally that the opal suppressor phosphoserine tRNA gene is located on chromosome 19. The flanking probe hybridized to a single homologous band in hamster and in mouse DNA to which the gene probe also hybridized, demonstrating that the 5'-flanking region of the opal suppressor tRNA gene is conserved in mammals. Restriction analysis of DNAs obtained from the white blood cells of 10 separate individuals demonstrates that the gene is polymorphic. This study provides two additional markers for the human genome and constitutes only the second set of two tRNA genes assigned to human chromosomes. PMID- 3038910 TI - Testosterone inhibits cAMP-induced de Novo synthesis of Leydig cell cytochrome P 450(17 alpha) by an androgen receptor-mediated mechanism. AB - The regulation of the novo synthesis of the microsomal cytochrome P-450 enzyme, P 450(17 alpha), was studied in mouse Leydig cell cultures. Chronic treatment with 0.05 mM 8-Br-cAMP (cAMP) caused a time-dependent increase in 17 alpha-hydroxylase activity and in the amount of P-450(17 alpha), quantitated by immunoblotting. This increase in both activity and amount was enhanced by inhibiting testosterone production with aminoglutethimide, an inhibitor of cholesterol side-chain cleavage or SU 10603, an inhibitor of 17 alpha-hydroxylase. To examine the mechanism by which cAMP or cAMP plus inhibitors of testosterone production increased the activity and amount of P-450(17 alpha), changes in the rate of de novo synthesis were studied by measuring [35S]methionine incorporation into newly synthesized protein. Treatment with cAMP plus aminoglutethimide or SU 10603 caused a 2-fold or greater increase in the rate of de novo synthesis of P-450(17 alpha) compared to treatment with cAMP only. The addition of exogenous testosterone reversed this increase in the rate of synthesis, indicating that testosterone modulates the extent of cAMP-stimulated induction of P-450(17 alpha). This negative effect of testosterone could be mimicked by the addition of the androgen agonist, mibolerone, and prevented by the addition of the antiandrogen, hydroxyflutamide. Neither estradiol nor dexamethasone had any effect on the synthesis of P-450(17 alpha). Studies on the degradation of newly synthesized P-450(17 alpha) demonstrated that testosterone had no effect on the decay of P-450(17 alpha) during the first 24 h but caused a significant increase in the rate of decay between 24 and 48 h. These data indicate that testosterone produced during cAMP induction of P-450(17 alpha) negatively regulates the amount of this cytochrome P-450 enzyme by two distinct mechanisms: by repressing cAMP induced synthesis of P-450(17 alpha) by an androgen receptor-mediated mechanism and by increasing the rate of degradation of P-450(17 alpha). A model is proposed for the regulation of P-450(17 alpha) in Leydig cells. PMID- 3038911 TI - Proximity and ligand-induced movement of interdomain residues in myosin subfragment 1 containing trapped MgADP and MgPPi probed by multifunctional cross linking. AB - The conformations of myosin subfragment 1 containing trapped MgADP or MgPPi have been studied by investigating the spatial disposition of the remainder of the subfragment 1 structure to the covalently bridged ATPase-related thiols SH1 and SH2. This has been done by synthesizing a trifunctional photoactivatable reagent 4,4'-bis(N-maleimido)benzophenone and reacting it with subfragment 1 in the presence of these ligands. Modification of subfragment 1 by this reagent mimics closely the changes in the ATPase properties as noted previously for modification with p-phenylenedimaleimide. In addition, noncovalent trapping of nucleotide also results, presumably by the bridging of the SH1 and SH2 thiols. On photolysis, cross-linking from the reagent bridging the thiols to other regions in subfragment 1 can be observed, but the extent and course of the photoinduced cross-linking depend on the nature of the trapped ligand. For subfragment 1 with trapped MgADP, a high efficiency cross-linking occurs between the 21-kDa segment and the 50-kDa segment. With MgPPi as the trapped ligand, low efficiency cross linking occurs between the bridged thiols and either the 27-kDa N-terminal or the 50-kDa segments of the heavy chain. These results indicate that without the adenosine moiety, the binding of MgPPi to subfragment 1 leaves the protein in a flexible state so that residues in both the 27-kDa and the 50-kDa segment can move within the cross-linking span of the activated benzophenone triplet. The trapping of MgADP apparently results in a more rigid state for the subfragment 1 in which residues in the 50-kDa segment are spatially close to the bridged thiols, thus enabling photocross-linking to proceed with higher efficiency. PMID- 3038912 TI - Identification of the bombesin receptor on murine and human cells by cross linking experiments. AB - The bombesin receptor present on the surface of murine and human cells was identified using 125I-labeled gastrin-releasing peptide as a probe, the cross linking agent disuccinimidyl suberate, and sodium dodecyl sulfate gels. A clone of NIH-3T3 cells which possesses approximately 80,000 bombesin receptors/cell with a single binding constant of approximately 1.9 X 10(-9) M was used in these studies. In addition, we used Swiss 3T3 cells and a human glioma cell line which possesses approximately 100,000 and approximately 55,000 bombesin receptors/cell, respectively. Under conditions found optimal for binding, it is demonstrated that 125I-labeled gastrin-releasing peptide can be cross-linked specifically to a glycoprotein of apparent molecular mass of 65,000 daltons on the surface of the NIH-3T3 cells. Similar results were obtained when the cross-linked product was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing or non-reducing conditions. Moreover, the cross-linking reaction is specific and saturable and the 65,000-dalton polypeptide is not observed when the cross-linking experiments were performed with a NIH-3T3 cell line which is devoid of bombesin receptors. Interestingly, glycoproteins with apparent molecular weights of 75,000 were labeled specifically by 125I-labeled gastrin-releasing peptide when similar experiments were performed with Swiss 3T3 cells and with human glioma cell line GM-340. These different molecular weights may indicate differential glycosylation as treatment with the enzyme N-glycanase reduced the apparent molecular weight of the cross-linked polypeptide to 45,000. On the basis of these results it is concluded that the cross-linked polypeptides represent the bombesin receptor or the ligand-binding subunit of a putative larger bombesin receptor expressed on the surface of these cells. PMID- 3038913 TI - Autophosphorylation of the alpha subunit of phosphorylase kinase from rabbit skeletal muscle. AB - The autophosphorylation of the alpha subunit of phosphorylase kinase occurs simultaneously at multiple sites during incorporation of the first mol of phosphate. The predominant and initial autophosphorylation site on this subunit is different than the major site phosphorylated by cAMP-dependent protein kinase, which also phosphorylates multiple sites, as evidenced by two-dimensional phosphopeptide maps. All of the sites on the alpha subunit phosphorylated by cAMP dependent protein kinase comigrate on peptide maps with autophosphorylation phosphopeptides; however, several phosphopeptides observed after autophosphorylation are not evident following phosphorylation by cAMP-dependent protein kinase. The phosphopeptide maps of the alpha subunit are the same whether autophosphorylation is carried out at pH 6.8 or 8.2 or whether MnATP is used instead of MgATP; there is only a slight difference in the maps brought about by EGTA-insensitive autophosphorylation. The autophosphorylation is shown to be an intrinsic activity of the phosphorylase kinase molecule; this conclusion is based on the observed copurification of the autophosphorylation activity with activities toward phosphorylase b and kappa-casein and the unaltered influence of various effectors on these activities throughout different sequential adsorption chromatography purification steps. Additional support to that already in the literature that the initial autophosphorylation events are predominantly intramolecular is gained by showing that previously autophosphorylated enzyme has little ability to catalyze the phosphorylation of nonphosphorylated enzyme. PMID- 3038914 TI - Sequence elements essential for rho-dependent transcription termination at lambda tR1. AB - To determine the location of DNA sequences required for the utilization of rho factor in transcription termination at the tR1 terminator of phage lambda, we constructed and analyzed a series of deletion mutants affecting several distinct regions of the cro-gene sequence. Two distinct sequence blocks, rutA and rutB, are shown to be particularly important for rho action at tR1 during in vitro transcription. They are located in the region between the sequence encoding the translation termination codon of cro mRNA and the start of tR1. Although other sequences contribute to rho action, their function can be replaced by unrelated heterologous DNA sequences encoding highly structured RNA segments, while the function of the rut sequences cannot. The lambda tR1 rut sequences encode RNA segments that contain a high proportion of cytidylate residues and that lack extensive intramolecular base pairing. Thus, the rut transcript segments are likely to be parts of the site on the nascent cro RNA that binds specifically with rho factor as an essential step in the termination process. This view is supported further by the findings (Chen, C.-Y. A., Galluppi, G. R., and Richardson, J. P. (1986) Cell 46, 1023-1028) that rho action at tR1 is specifically inhibited by DNA oligonucleotides that form hybrid helices with rut transcript segments. We also examined the effect of the auxiliary transcription factor NusA on transcription of our mutant templates and show that deletion of the boxA sequence of cro gene does not diminish the ability of NusA to reduce rho action at tR1. PMID- 3038915 TI - A native cruciform DNA structure probed in bacteria by recombinant T7 endonuclease. AB - T7 endonuclease preferentially cleaves purified supercoiled pBR322 and colE1 plasmids at the single-stranded regions exposed when palindromic sequences assume cruciform structures (Panayotatos, N., and Wells, R.D. (1981) Nature 289, 466 470). In vivo, however, induction of nuclease synthesis off a cloned gene caused complete degradation of the bacterial DNA but not of the plasmid vector; presumably, single-stranded regions (cruciforms?) on the genome effectively complete for the nuclease with similar sites on the plasmid (Panayotatos, N., and Fontaine, A. (1985) J. Biol. Chem. 260, 3173-3177). To overcome this competition, we introduced on the plasmid the naturally occurring colE1 palindrome which forms a more stable cruciform in vitro. In addition, we increased the target size (and the T7 endonuclease gene dosage) by raising the copy number of the plasmid 5 fold. Induction of the endonuclease encoded by this new plasmid (pLAT75) resulted not only in degradation of genomic DNA but also in intracellular nicking and linearization of the plasmid. The cleavage site in vivo was mapped at the colE1 palindrome and coincided with the site cleaved specifically in vitro by either T7 or S1 endonuclease only when this palindrome assumes the cruciform structure. These results indicate that cruciform structures exist intracellularly and demonstrate the usefulness of endonucleases as probes of DNA topology in vivo. PMID- 3038916 TI - ELISA for bovine herpesvirus 1 (BHV1) antibody with HIRT supernatant. AB - Detailed studies were made of the ability of HIRT supernatant (HS) from bovine herpesvirus 1 (BHV1) infected cultures to sensitize plates for enzyme-linked immunosorbent assay (ELISA). The ultracentrifuged pellet of HS had less sensitizing activity than the supernatant, but the antigen was removed completely by 0.22 micron filters and to some extent by 0.45 micron filters; it was minimally affected by sonication; it was destroyed by the action of Pronase but not by DNAase I when a kallikrein inactivator was added and the mixture incubated; incubation with DNAase II had no effect. Thus the presence of DNA was not required for the sensitizing activity of HS and the antigens recognized by antibodies in HS in ELISA were directed to its protein component. Strong reactions were given in immunoblotting of HS from BHV1 infected tissue cultures with anti-BHV1 glycoprotein monoclonal IgG, but HS from uninfected tissue cultures did not react with the same monoclonal IgG. PMID- 3038917 TI - Peripheral effects of neurokinins: functional evidence for the existence of multiple receptors. AB - The ability of six neurokinins (substance P, neurokinin A, neurokinin B, physalaemin, eledoisin and kassinin) to induce hypotension, salivary secretion and to activate motility of the duodenum and of the urinary bladder was investigated in urethane-anaesthetized rats. A comparison of the relative potency of these substances in producing a given biological effect yielded results consistent with the hypothesis of the existence of three distinct types of receptor in rat peripheral tissues, namely SP-P, SP-K and SP-E according to the nomenclature proposed by Buck, Burcher, Shults, Lovenberg & O'Donohue (1984), or NK-P, NK-A and NK-B according to that of Regoli, D'Orleans-Juste, Drapeau, Dion & Escher (1985). An NK-P receptor is responsible for the production of the hypotensive effect and for the activation of salivary secretion. An 'NK-P like' receptor subtype may be involved in determining the direct contractile effects on muscle cells of neurokinins in the rat isolated urinary bladder and the 'phasic' contraction of the rat duodenum. An NK-A receptor mediates the 'tonic' contraction of the rat duodenum while an NK-B receptor mediates the activation of the micturition reflex. Evidence is presented that multiple neurokinin receptors are present in the same organ and participate with different modalities to the regulation of smooth muscle function. PMID- 3038918 TI - Increased beta 2-adrenoreceptor density in heart, kidney and lung of spontaneously hypertensive rats. AB - The development of beta-adrenoreceptor density and beta 1- and beta 2 adrenoreceptor distribution has been investigated in heart, kidney and lung of spontaneously hypertensive (SHR) and normotensive (WKY) rats by (-)125I iodocyanopinolol binding at the age of 5-6, 9-10 and 19-21 weeks. At all ages beta-adrenoreceptor density was similar in hearts of both strains, while it was increased in kidney and lung of SHR compared to WKY. The beta 2-adrenoreceptor density was higher in all three tissues of SHR at all ages investigated. On the other hand, beta 1-adrenoreceptor density was decreased in heart, unchanged in lung and increased in kidney of SHR compared to WKY. Destruction of presynaptic nerve terminals by treatment of WKY with 6-hydroxydopamine produced a 24% loss of cardiac beta 2-adrenoreceptors, whereas the beta 1-adrenoreceptor density remained unchanged, suggesting that at least part of the cardiac beta 2 adrenoreceptor population is localized prejunctionally. It is suggested that beta 2-adrenoreceptors are involved in the development or maintenance of high blood pressure in SHR, possibly by facilitating noradrenaline release. PMID- 3038919 TI - Adherence of Streptococcus mutans to implant materials. AB - The adherence of Streptococcus mutans OMZ-176 to six implant materials (poly crystal alumina, single-crystal alumina, titanium alloy, cobalt-chromium molybdenum alloy, hydroxyapatite, and heat-curing polymethylmethacrylate resin) was studied in vitro. The change of free energy, which corresponds to the adherence process, was also evaluated for hydrophobic interaction. Adherence of S. mutans to poly-crystal alumina was lower than adherence to other materials. The adherence of S. mutans to the test materials was highly correlated with the change of free energy, which suggests that hydrophobic interaction plays an important role in the adherence of S. mutans to the implant materials. PMID- 3038920 TI - Paralysis of the posterior interosseous nerve caused by tumour: brief report. PMID- 3038921 TI - Leukotrienes LTB4 and LTC4 in thermally injured patients' plasma and burn blister fluid. AB - Radioimmunoassays for 5(S), 12(R)-dihydroxyeicosa-6, 14-cis, 8,10-trans tetraenoic acid (leukotriene B4, LTB4) and 5(S)-hydroxy-6(R)-L-gamma-glutamyl-L cysteinyl-glycinyleicosa-7,9- trans, 11,14-cis-tetraenoic acid (leukotriene C4, LTC4) have been used to quantify the concentrations of these arachidonic acid metabolites in plasma (12 patients) and blister fluid (6 patients) of burned patients. The results of this study demonstrate that, in general, burn plasma LTB4, and LTC4 were not elevated; however, in individual cases, transient high levels of leukotrienes were observed. Correlation between leukotriene "peaks" and the clinical course could not be established. High levels of LTB4 and LTC4 in burn blister fluid suggest their participation in local inflammatory reactions. PMID- 3038922 TI - The three-dimensional structure of the Na,K-ATPase from electron microscopy. AB - The structure of Na,K-ATPase has been studied by electron microscopy and image reconstruction. A three-dimensional structure of this enzyme has been obtained to an overall resolution of 2.5 nm using data from specimens of negatively stained dimer sheets tilted through a range of angles +/- 60 degrees. The reconstruction shows a complex mass distribution consisting of ribbons of paired molecules extending approximately 6.0 nm from the cytoplasmic side of the membrane. The molecular envelope consists of a massive "body" with "lobe" and "arm" structures projecting from it. The body has a columnar shape and is tilted with respect to the plane of the membrane. The region of interaction responsible for dimer formation is located between two bodies and is clearly visible in the reconstruction. It has been identified as a segment in the amino-terminal portion of the alpha subunit. The arms that interconnect the ribbons are located close to the membrane and are most probably formed by the beta subunits. PMID- 3038923 TI - Two distinct attachment sites for vimentin along the plasma membrane and the nuclear envelope in avian erythrocytes: a basis for a vectorial assembly of intermediate filaments. AB - In vitro binding studies with isolated bovine lens vimentin and avian erythrocyte membranes reveal the existence of two functionally distinct sets of intermediate filament attachment sites. One population of such receptors is located along the nuclear envelope and comprises polypeptides recognizing the carboxy-terminal tail domain of vimentin. Vimentin associates with these nuclear surface receptors in a cooperative manner and forms extensive 10-nm filaments in a concentration dependent fashion. Conversely, the plasma membrane contains binding sites that interact in a noncooperative, saturable fashion with vimentin, recognizing its amino-terminal head domain. The functional dichotomy of the vimentin-binding sites under in vitro conditions may reflect a vectorial assembly process whereby 10-nm filaments, although structurally apolar, acquire polar features brought about by the differential attachment to specific receptors arranged along the plasma membrane and the nuclear envelope. PMID- 3038924 TI - An antibody specific for an endoproteolytic cleavage site provides evidence that pro-opiomelanocortin is packaged into secretory granules in AtT20 cells before its cleavage. AB - We have raised a rabbit antiserum against a synthetic peptide corresponding to the cleavage site between beta-lipotropic hormone and the ACTH moieties of murine pro-opiomelanocortin (POMC). After affinity purification, the anti-cleavage site antibody immunoprecipitates POMC from extracts of AtT20 cells but it does not immunoprecipitate the ACTH in such extracts or any of the other products of cleavage of POMC. By contrast, an antiserum raised against pure swine ACTH immunoprecipitates both POMC and ACTH from AtT20 cell extracts. Using the anti cleavage site antibody we have shown that all the POMC synthesized during a 15 min pulse-labeling with [35S]methionine is cleaved at this site within 1 h. By immunoelectron microscopy we show that approximately 25-30% of peripheral secretory granules in AtT20 cells can be labeled with the anti-cleavage site antibody while anti-ACTH antiserum labels all these granules. This establishes that at least some POMC is packaged into secretory granules before its proteolytic cleavage. PMID- 3038925 TI - The endosomal concentration of a mannose 6-phosphate receptor is unchanged in the absence of ligand synthesis. AB - The cation-independent mannose-6-phosphate (Man-6-P) receptor is involved in the targeting of newly synthesized lysosomal hydrolases. To investigate the intracellular distribution of this receptor, a conjugate of lactoperoxidase coupled to asialoorosomucoid was used to catalyze its iodination within the endosomes of human hepatoma (HepG2) cells. The 215-kD, cation-independent Man-6-P receptor was iodinated by this procedure as shown by pentamannosyl-6-phosphate Sepharose affinity chromatography and by immunoprecipitation of labeled cell extracts. The amount of this receptor detected in endosomes was found to be unchanged after inhibition of protein synthesis with cycloheximide. If the Man-6 P receptor accumulates in the Golgi apparatus in the absence of lysosomal hydrolase synthesis, it should have been correspondingly depleted from endosomes after a period of cycloheximide treatment, because these pools of receptor are in rapid equilibrium. Therefore, these data suggest that newly synthesized ligands are not required for the transport of the cation-independent Man-6-P receptor from the Golgi apparatus to endosomes. PMID- 3038926 TI - Calcium-independent contraction in lysed cell models of teleost retinal cones: activation by unregulated myosin light chain kinase or high magnesium and loss of cAMP inhibition. AB - The retinal cones of teleost fish contract at dawn and elongate at dusk. We have previously reported that we can selectively induce detergent-lysed models of cones to undergo either reactivated contraction or reactivated elongation, with rates and morphology comparable to those observed in vivo. Reactivated contraction is ATP dependent, activated by Ca2+, and inhibited by cAMP. In addition, reactivated cone contraction exhibits several properties that suggest that myosin phosphorylation plays a role in mediating Ca2+-activation (Porrello, K., and B. Burnside, 1984, J. Cell Biol., 98:2230-2238). We report here that lysed cone models can be induced to contract in the absence of Ca2+ by incubation with trypsin-digested, unregulated myosin light chain kinase (MLCK) obtained from smooth muscle. This observation provides further evidence that MLCK plays a role in regulating cone contraction. We also report here that lysed cone models can be induced to contract in the absence of Ca2+ by incubation with high concentrations of MgCl2 (10-20 mM). Mg2+-induced reactivated contraction is supported by inosine triphosphate (ITP) just as well as by ATP. Because ITP will not serve as a substrate for MLCK, this finding suggests that Mg2+-activation of contraction does not require myosin phosphorylation. Although Ca2+-induced contraction is completely blocked by cAMP at concentrations less than 10 microM, cAMP has no effect on cone contraction activated by unregulated MLCK or by high Mg2+ in the absence of Ca2+. Because trypsin digestion of MLCK cleaves off not only the Ca2+/calmodulin-binding site but also the site phosphorylated by cAMP-dependent protein kinase, and because Mg2+ activation of cone contraction circumvents MLCK action altogether, both these observations would be expected if cAMP inhibits reactivated cone contraction by catalyzing the phosphorylation of MLCK and thus reducing its affinity for Ca2+, as has been described for smooth muscle. Together our results suggest that in lysed cone models, myosin phosphorylation is sufficient for activating cone contraction, even in the absence of other Ca2+ mediated events, that cAMP inhibition of contraction is mediated by cAMP dependent phosphorylation of MLCK, and that 10-20 mM Mg2+ can activate actin myosin interaction to produce contraction in the absence of myosin phosphorylation. PMID- 3038927 TI - Cell surface dynamics of neutrophils stimulated with phorbol esters or retinoids. AB - Neutrophils undergo rapid morphological changes as well as metabolic perturbations when stimulated with certain phorbol esters. Stimulated cells initially exhibit pronounced projections emanating from the cell bodies, followed by rounding of the cells, reduction in granule number, and the appearance of intracellular vesicles. We show these vesicles to be derived, at least in part, from the plasmalemma. The experimental approach involved labeling stimulated and unstimulated cells with native ferritin and cationized ferritin, along with the cytochemical localization of ecto-5'-nucleotidase. The labeling patterns of the vesicles indicate that these structures are involved in both phorbol ester stimulated adsorptive and fluid-phase endocytosis. Neutrophils stimulated with 12 O-tetradecanoyl-phorbol-13-acetate (TPA) exhibit two distinct rates of superoxide release in which the second, prolonged level is approximately 50% of the initial rate. All-trans-retinal, which we have recently shown to stimulate O2- release but not granule exocytosis or cell vesiculation, induces a single prolonged rate of maximal O2- release. Neutrophils treated with both all-trans-retinal and TPA exhibit only a single sustained rate of maximal O2- release similar to that observed with all-trans-retinal alone. Moreover, treatment of cells with all trans-retinal blocks the vesiculation of neutrophils induced by TPA in a dose dependent manner. This observation provides a possible explanation for the differences in the kinetics of superoxide release. PMID- 3038928 TI - Modulation of fibronectin gene expression in chondrocytes by viral transformation and substrate attachment. AB - Chicken vertebral chondrocytes, which normally grow in suspension, synthesize large amounts of cartilage extracellular matrix proteins, but little fibronectin. We have analyzed the effects of both substrate attachment and transformation with a temperature-sensitive mutant of Rous sarcoma virus on fibronectin gene expression in these cells. Our experiments show that viral transformation increases fibronectin synthesis to a greater extent than substrate attachment. Furthermore, transformed chondrocytes have lost the ability to decrease fibronectin synthesis in response to suspension culture, suggesting that transformation alters the normal attachment-responsive control of fibronectin gene expression. Finally, infected substrate-attached chondrocytes shifted to the nonpermissive temperature for transformation use fibronectin RNA more efficiently in protein synthesis than cells grown under the other conditions, suggesting for the first time a role for translational control of fibronectin gene expression. PMID- 3038929 TI - pH-dependent function, purification, and intracellular location of a major collagen-binding glycoprotein. AB - A major collagen-binding heat shock protein of molecular mass 47,000 D was found to bind to collagen by a pH-dependent interaction; binding was abolished at pH 6.3. Native 47-kD protein could therefore be purified from chick embryo homogenates in milligram quantities by gelatin-affinity chromatography and gentle acidic elution. Rat monoclonal and rabbit polyclonal antibodies were generated against the purified 47-kD protein. Immunofluorescence microscopy of cultured chick embryo fibroblasts with these antibodies revealed bright, granular perinuclear staining as well as a weaker reticular network structure towards the cell periphery, suggesting that this protein was located in the endoplasmic reticulum. No immunofluorescence staining was detected on the cell surface. Double-staining experiments with these antibodies and fluorescently labeled wheat germ agglutinin suggested that the 47-kD protein was absent from the Golgi apparatus. Localization of the 47-kD protein in the endoplasmic reticulum but not in the Golgi complex was confirmed by immunoelectron microscopy. In vivo localization studies using immunohistochemistry of cryostat sections of chick liver revealed that the 47-kD protein was present in fibrocytes, Kupffer cells, and smooth muscle cells. It was absent from hepatocytes and the epithelia of bile ducts or sinusoidal endothelium. This major transformation- and heat shock regulated glycoprotein is thus localized intracellularly, is expressed in only certain cells, and functions in a pH-regulated manner. These findings suggest that this glycoprotein is not likely to be a general cell-surface collagen receptor, but may instead play roles in intracellular protein processing or translocation. PMID- 3038932 TI - Neuroreceptor assay with positron emission tomography: definition of transfer coefficients. PMID- 3038931 TI - Effects of mutations in three domains of the vesicular stomatitis viral glycoprotein on its lateral diffusion in the plasma membrane. AB - The lateral mobility of the vesicular stomatitis virus spike glycoprotein (G protein) and various mutant G proteins produced by site-directed mutagenesis of the G cDNA has been measured. Fluorescence recovery after photobleaching results for the wild type G protein in transfected COS-1 cells yielded a mean diffusion coefficient (D) of 8.5 (+/- 1.3) X 10(-11) cm2/s and a mean mobile fraction of 75% (+/- 3%). Eight mutant proteins were also examined: dTM14, lacking six amino acids from the transmembrane domain; TA2, lacking an oligosaccharide in the extracellular domain; QN2, possessing an extra N-linked oligosaccharide in the extracellular domain; CS2, possessing a serine instead of a cysteine at residue 489 in the cytoplasmic domain, preventing palmitate addition to the glycoprotein; TMR-stop, lacking the entire cytoplasmic domain except an arginine at residue 483; and three chimeric proteins, G mu, G23, and GHA, containing in place of the 29 amino acid wild type cytoplasmic domain the cytoplasmic domains from the surface IgM from the spike protein of the infectious bronchitis virus or from the hemagglutinin protein of the influenza virus, respectively. The mean D for the mutant proteins varied over a relatively small range, with the slowest mutant, G23, exhibiting a value of 11.3 (+/- 1.4) X 10(-11) cm2/s and the fastest mutant, GHA, having a D of 28.6 (+/- 4.5) X 10(-11) cm2/s. The mean mobile fraction similarly varied over a small range, extending from 55 to 68%. None of the mutations resulted in the more rapid diffusion characteristic of membrane proteins embedded in artificial bilayers. Therefore, it appears that the cytoplasmic and transmembrane domains themselves contribute little to restraining the lateral mobility of this integral membrane protein when expressed in transfected cells. PMID- 3038930 TI - Two distinct cell-binding domains in laminin can independently promote nonneuronal cell adhesion and spreading. AB - Two subfragments of laminin, E8, a major part of the long arm, and E1-4, the three short arms, promote cell adhesion and spreading. Three distinct types of adhesive behavior are seen in short term (1 h) assays, typified by secondary murine fibroblasts, adherent only on fibronectin; secondary murine myoblasts, adherent on fibronectin, laminin, and the E8 fragment; and Rugli human glioblastoma cells, adherent on fibronectin, laminin, E8, and E1-4. E8-specific polyclonal antibodies block myoblast adhesion to E8 and to laminin with identical concentration dependence; Rugli binding to E8 but not to laminin is also totally blocked by these antibodies. Heating of E8 and laminin to approximately 60 degrees C abolishes cell attachment-promoting activity for myoblasts. Adhesion of Rugli cells to E8 is also lost, but on laminin the attachment-promoting activity remains constant. This is due to an increase in the activity of E1-4 fragment as it is heated. Thus, major sites for initial cell adhesion to and spreading on laminin lie within the E8 and E1-4 fragments, but not all cells binding to laminin will bind to both fragments. These data may tentatively be explained by the existence of more than one type of receptor for laminin at the cell surface; one is needed for each fragment. PMID- 3038934 TI - Measurement of busulfan in plasma by high-performance liquid chromatography. PMID- 3038933 TI - [Ruptured adenoma of the liver with severe pulmonary embolism discovered during a cesarean section]. AB - A ruptured adenoma of liver was detected during a caesarean in a multipara woman at full term with a severe pulmonary embolus. Therapy for this rare and curious case, with combined internal hemorrhage, severe pulmonary embolism and full term pregnancy in a multipara, is discussed. PMID- 3038935 TI - Detection of HIV envelope specific antibodies by immunoelectron microscopy and correlation with antibody titer and virus neutralizing activity. AB - Human antisera positive for HIV were evaluated on HTLV-IIIB producing cells by two different immunoelectron microscopic (IEM) techniques. In preembedding immunoferritin IEM a heavy label was observed with early budding HIV. Under the same conditions cell released 'mature' particles were almost negative, which could be explained by the direct observation that most of the surface glycoprotein knobs are lost spontaneously during virus maturation. Using freshly infected cultures after agarose embedding, immunogold labelling of ultrathin cryosections allowed us to detect and differentiate internal core as well as virus envelope antigens. A good qualitative correlation between neutralization titers and IEM labelling intensity was observed. This type of immunocryoultramicrotomy appears to be useful for the detection of antigens in and on the virion. It might turn out valuable for the characterization of the env gp120 epitopes of HIV. PMID- 3038936 TI - Evaluation of a commercial latex agglutination test for detecting antibodies to cytomegalovirus in organ donors and transplant recipients. AB - A commercial latex agglutination test was evaluated for use in detecting CMV antibody in organ donors and transplant recipients. When compared with indirect and competitive enzyme-linked immunosorbent assays (ELISAs) and complement fixation tests (CFT), the test gave concordant results with 96.6% of sera. The latex agglutination test was more sensitive than CFT but less sensitive than ELISA. The major advantage of the latex agglutination test was its simplicity and rapidity. Taking only 8 min to perform, it is very suitable for testing sera from organ donors, since the time available for such tests is short because of the finite ischaemic time, particularly for liver and heart transplantation. PMID- 3038937 TI - BK virus in cell culture: infectivity quantitation and sequential expression of antigens detected by immunoperoxidase staining. AB - We applied an indirect immunoperoxidase staining method for the detection of BK virus (BKV) antigens in Vero cells. By immunoperoxidase staining of sequential samples, expression of BKV T- and structural antigens was first detectable 72 h postinfection (pi). The initial low number of antigen positive cells was slowly increasing until 6 d pi. Thereafter, a steady progression was observed until total cytopathogenic effect 30 d pi. Viral hemagglutinins were detected 11 d pi in both the culture medium and in the cell lysate. Passage of the preparations into fresh Vero cells and immunoperoxidase detection demonstrated the presence of infectious virions in both the cell lysates and the medium from 2 d pi. The number of infectious particles in both media and cell lysates correlated to the number of IP stained cells in the sequential staining. We suggest that this type of assay might be used to examine the effects, in vitro, of interferons and other virostatic drugs of BKV and other viruses. PMID- 3038939 TI - Effect of glutaraldehyde on the antigenicity and infectivity of hepatitis A virus. AB - The effect of glutaraldehyde on the antigenicity and infectivity of hepatitis A virus (HAV) was examined. The CF 53 strain, adapted to human hepatoma PLC/PRF/5 cells, was treated with glutaraldehyde using three different concentrations, 0.02, 0.10, and 0.50%, for various periods of time, 3, 10, and 30 min, respectively. After the virucidal assays, glutaraldehyde and HAV were separated by gel filtration, then the antigen (radioimmunoassay) titer and the infectivity titer were determined. The greatest antigen titer reduction was about 80% after 30 min using 0.10% glutaraldehyde and within only 3 min using 0.50% glutaraldehyde. Glutaraldehyde is an effective disinfectant against HAV: the infectious virus titer decreased by more than 3 log10 after 30 min using 0.10% glutaraldehyde and within only 3 min using 0.50% glutaraldehyde. Statistical studies showed that the decrease of antigen or infectious virus titer was affected by both glutaraldehyde concentration and exposure time. PMID- 3038938 TI - Quantitative assay of recombinant vaccinia virus-encoded neomycin phosphotransferase in infected eukaryotic cell lysates. AB - A method for the detection and quantitation of neomycin phosphotransferase (NPT II) activity in recombinant vaccinia virus (VV)-infected eukaryotic cell lysates is described. The assay is linear with respect to both protein concentration and time of incubation. Cytoplasmic extracts of cells infected with a recombinant VV expressing the bacterial neo gene exhibited NPT II levels more than 50-fold higher than those detected in extracts from either uninfected or VV-infected cells. These results indicate that interference from cellular or viral-induced ATPase activities is sufficiently low that NPT II enzyme activity can be measured in crude cell lysates without employing additional protein purification procedures. PMID- 3038940 TI - Detection of CMV in urine: comparison between DNA-DNA hybridization, virus isolation, and immunoelectron microscopy. AB - Rapid detection of CMV-DNA in urine specimens by dot-blot hybridization was compared to conventional virus isolation and to virus identification using solid phase immunoelectron microscopy (SPIEM). To detect viral DNA, 32P-labeled EcoR1 J fragment of CMV-DNA was used as a probe in the hybridization assay. In addition, DNA extracted from infected human embryo fibroblasts (amplified DNA) was also hybridized to the same probe. Urine specimens were obtained from 10 renal transplanted patients, seven premature infants, three family members, and five children suspected of CMV infection. CMV was isolated from 10 urine specimens and SPIEM detected viral particles in nine specimens. Ten positive samples were identified as such by hybridization with DNA extracted directly from urine specimens, while hybridization with amplified DNA yielded 17 positives. Only in one urine specimen, positive by virus isolation and SPIEM, DNA was not detected by the hybridization assays. Elevated IgG or IgM-specific antibodies were found in 10 patients. Hybridization with amplified DNA proved the most sensitive and relatively rapid assay, as compared with direct DNA detection in urine, tissue culture isolation, SPIEM, or serologic tests. PMID- 3038941 TI - An assessment of the sensitivity of three methods for the detection of rotavirus. AB - A comparison was made of the sensitivity of a current commercial ELISA for detecting rotavirus in faecal specimens with the more complex, technically demanding systems of electron microscopy and polyacrylamide gel electrophoresis. Even after modification, the ELISA failed to detect 22% of specimens with particles identifiable by electron microscopy. Polyacrylamide gel electrophoresis failed to identify 2 out of 50 specimens with particles present but did distinguish 2 group C rotaviruses. PMID- 3038942 TI - Rapid detection of cytomegalovirus using immune scanning electron microscopy. AB - Monoclonal antibodies specific for human cytomegalovirus were conjugated to latex microspheres that were already labelled with rabbit anti-mouse immunoglobulin. The beads were incubated with serum or urine from patients, and then collected on a filter surface, which was analyzed in a scanning electron microscope. Size, immunological specificity, and relative quantity of virus particles were determined within 2 h after serum or urine collection by the visualization of virus particles specifically bound to the latex bead surface. No such binding of virus particles were detected in the various controls. This method was compared with conventional virus isolation by tissue culture. It enables identification of viruses within a few hours in different body fluids. Even without specific antibodies, the filtration method may permit the rapid detection of particles and the determination of their size in various body fluids. PMID- 3038943 TI - Differences in outcome between black and white elderly hip fracture patients. AB - Because blacks make up a small proportion of the hip fracture population, little is known about how blacks' experience following hip fracture compares to that of whites. This study, a retrospective review of the medical records of 119 community residing subjects, 60 years of age and older, 37% of whom were black, admitted with a diagnosis of hip fracture to a large urban teaching hospital, investigated differences between black and white patients in factors associated with outcome following hip fracture and outcomes at time of hospital discharge. Blacks were significantly more likely than whites to exhibit a high total number of diagnoses, urinary incontinence following surgery, low admission hemoglobins and mental impairment. Blacks experienced significantly longer hospitalizations than whites, were significantly more likely to be nonambulatory at discharge and showed different patterns of discharge destination. Multiple regression and logistic regression indicated that the greater amount of illness of blacks is a consistent significant predictor of these differences in outcomes, but that race, delays to surgery and non-surgical treatment also make independent significant contributions. These findings indicate the importance of planning for the in hospital care of black hip fracture patients, and the examination of the financial consequences of DRG's for hospitals serving black or poor populations. PMID- 3038944 TI - Reported herpes-virus-infection, fever and cancer incidence in a prospective study. AB - As soon as the first clinico-epidemiology reports appeared describing an association of herpes virus infection with genital cancer, relevant questions were introduced into an ongoing follow-up of a cohort already being studied for psychosocial risk factors in cancer. The study, which was initiated in 1966, comprised 1353 persons; disease incidence was determined in 1976. Incidences of cancer (including extragenital cancer) were found to be directly related to the reported prevalence of genital herpes symptoms, but were related inversely to extragenital herpes prevalence and to previous fever episodes. With regard to the relatively common incidence of "fever blisters", i.e. facial herpes, attributed to HSV-type 1, the latter findings is less surprising than the strong association of HSV type 2 lesions with overall cancer incidence. This association was particularly noticeable in individuals who had not reported episodes of fever earlier. These findings indicate the predictive role of certain host factors which are possibly related to systematic responses of the organism, including a diminished protection against (latent) herpes genitalis infection (recurrences). Further prospective studies are needed to investigate the proper time relationship between clinically-overt herpes genitalis and the manifestation of cancer and its possible role in provoking the recurrent eruption of latent infections. PMID- 3038945 TI - Interleukin-2 effects on human B cells activated in vivo. AB - While activated B cells, as well as T cells, can express functional interleukin-2 receptors (IL-2R), the physiologic role of IL-2 in B-cell responses is still in doubt. Accordingly, we have examined the role of recombinant IL-2 (rIL-2) in the proliferative response, IL-2R expression, and the terminal differentiation of tonsillar B cells in comparative studies with T cells from the same source. For these analyses of physiologically activated lymphocytes, the B and T cells were first separated, then divided into subpopulations enriched for either resting or preactivated cells on the basis of relative cell density or activation antigen expression. As expected, the relatively low-density fraction of T cells was enriched for IL-2R-positive (Tac+) cells and displayed vigorous proliferative responses to IL-2 along with heightened Tac antigen expression. Moreover, limiting dilution analysis revealed that growth of these activated T cells could be perpetuated with the addition of IL-2. By comparison, relatively few tonsillar B cells expressed the Tac antigen. The addition of rIL-2 to cultured B cells of relatively low density resulted in an increase in Tac+ cells but only a minimal proliferative response. The subpopulation of tonsillar B cells expressing the Bac 1 activation antigen contained most of the IL-2 inducible Tac+ B cells, and rIL-2 induced an efficient but transient proliferative response by these activated B cells. The rIL-2-induced Tac+ B cells were noted to be relatively large. A fraction of these Tac+ cells, but not the Tac- cells, produced and secreted immunoglobulins. Incubation with rIL-2 enhanced the Ig secretion, and the anti Tac antibody blocked this enhancement. Time-course analysis revealed that rIL-2 induced transient Tac expression, whereas mature plasma cells in 6-day cultures no longer expressed detectable Tac antigen. In conclusion, these observations suggest that IL-2 transiently upregulates expression of IL-2R, via which it induces the terminal growth and differentiation of activated B cells into plasma cells. PMID- 3038946 TI - Two-year survey of etiologic agents of diarrheal disease at San Lazaro Hospital, Manila, Republic of the Philippines. AB - The prevalence of bacterial pathogens and rotavirus in 2,908 patients with diarrhea who were admitted to San Lazaro Hospital in Manila in 1983 and 1984 was determined. One or more enteric pathogens were isolated or detected in samples from 1,698 (58.4%) patients. Isolation rates for the various enteropathogens were as follows: rotavirus, 30.6%; Shigella spp., 11.6%; Salmonella spp., 9.2%; enterotoxigenic Escherichia coli (1983 only), 7.8%; Vibrio cholerae biotype eltor, 3.8%; non-O1 V. cholerae, 2.8%; Vibrio parahaemolyticus, 1.7%; other Vibrio spp., 1.1%; Campylobacter jejuni, 3.0%; Aeromonas hydrophila, 1.3%; and Plesiomonas shigelloides 1.1%. Giardia lamblia and Entamoeba histolytica were detected in 0.6 and 0.1%, respectively, of stool samples examined. Determination of the etiologic role of isolates was complicated by one or more of the following factors: isolation of multiple enteric pathogens (302 cases); isolation of Salmonella spp., enterotoxigenic E. coli, and C. jejuni from a similar proportion of asymptomatic control patients and patients with diarrhea; and isolation of a high proportion of certain pathogens (especially Salmonella spp.) only from enrichment broth, suggesting infection with a small number of organisms. Isolation of V. cholerae eltor was seasonal, with the majority of cases occurring in the rainy months. In addition, the number of patients with diarrhea increased with the onset of the monsoon rains and peaked during the months of maximum rainfall. Rotavirus infection occurred in both children and adults throughout the year and was the most frequently identified cause of diarrhea in children under 5 years of age. Shigella spp. were the most common agents of diarrhea in adults. PMID- 3038947 TI - Genetic analysis of a human rotavirus that belongs to subgroup I but has an RNA pattern typical of subgroup II human rotaviruses. AB - We have previously found (O. Nakagomi, T. Nakagomi, H. Oyamada, and T. Suto, J. Med. Virol. 17:29-34, 1985), during an epidemiological study in Japan, a novel human rotavirus that belongs to subgroup I but has a long RNA pattern typical of subgroup II human rotaviruses. From the stool specimen containing this virus, we successfully isolated in MA104 cells a rotavirus, designated AU-1, which possesses these novel characteristics. The possibility that strain AU-1 was a laboratory contaminant of an animal rotavirus previously adapted to tissue culture cells was ruled out, and the identity of the AU-1 strain was established. Genetic analysis by RNA-RNA hybridization revealed that the AU-1 strain is not a simple reassortant between subgroup I and II human rotaviruses but that it shares a high level of sequence homology only with the gene encoding VP7 (the major neutralization protein) of serotype 3 human rotaviruses. Weak homology of the genomic RNA segments was also observed between the AU-1 strain and animal rotavirus strains, including rhesus rotavirus strain RRV and bovine rotavirus strain NCDV. These results suggest that the AU-1 strain may be an animal rotavirus that infected a human. PMID- 3038948 TI - Genetic relatedness among human rotavirus genes coding for VP7, a major neutralization protein, and its application to serotype identification. AB - Antigenic characterization of human rotaviruses by plaque reduction neutralization assay has revealed four distinct serotypes. The outer capsid protein VP7, coded for by gene 8 or 9, is a major neutralization protein; however, studies of rotaviruses derived from genetic reassortment between two strains have confirmed that another outer capsid protein, VP3, is in some cases equally important in neutralization. In this study, the genetic relatedness of the genes coding for VP7 of human rotaviruses belonging to serotypes 1 through 4 was examined by hybridization of their denatured double-stranded genomic RNAs to labeled single-stranded mRNA probes derived from human-animal rotavirus reassortants containing only the VP7 gene of their human rotavirus parent. A high degree of homology was demonstrated between the VP7 genes of strain D and other serotype 1 human rotaviruses, strain DS-1 and other serotype 2 human rotaviruses, strain P and other serotype 3 human rotaviruses, and strain ST3 and other serotype 4 human rotaviruses. Hybrid bands could not be demonstrated between the VP7 gene of D, DS-1, P, or ST3 and the corresponding gene of human rotaviruses belonging to a different serotype. RNA specimens extracted from the stools of 15 Venezuelan children hospitalized with rotavirus diarrhea were hybridized to each of the reassortant probes representing the four human serotypes. All five viruses with short RNA patterns showed homology with the DS-1 strain VP7 gene; two of these were previously adapted to tissue culture and shown to be serotype 2 strains by tissue culture neutralization. Of the remaining 10 viruses with long RNA patterns, 2 hybridized only to the D strain VP7 gene, 6 hybridized only to the P strain VP7 gene, and 2 hybridized only to the ST3 strain VP7 gene. Hybridization using single human rotavirus gene substitution reassortants as probes may provide an alternative method for identifying the VP7 serotype of field isolates that would circumvent the need for tissue culture adaptation. PMID- 3038949 TI - Serial propagation of porcine group C rotavirus (pararotavirus) in primary porcine kidney cell cultures. AB - A porcine group C rotavirus was adapted to serial propagation in roller tube cultures of primary porcine kidney cells with high concentrations of pancreatin. Infected cells were identified by immunofluorescence staining of cell monolayers. Only group C rotavirus particles were observed in culture supernatants by immune electron microscopy. PMID- 3038950 TI - Saliva as a source of feline leukemia virus antigen for diagnosis of disease. AB - An enzyme-linked immunosorbent assay (ELISA) for detection of feline leukemia virus (FeLV) p27 in saliva was tested for its accuracy and sensitivity in diagnosing FeLV infections. Saliva and serum samples from 564 clinical cases were tested with a 99.2% specificity. The overall accuracy of the saliva ELISA reactive to the serum ELISA was 97.9%. Experimentally, the ELISA saliva was the least sensitive in diagnosing early FeLV infections. However, the overall accuracy, ease of use, and simplicity of the test support its use as a screening procedure in clinical practice. PMID- 3038951 TI - Evaluation of three types of cell culture for recovery of adenovirus from clinical specimens. AB - The performance characteristics of HEK, HDF, and MK cells for adenovirus isolation were examined for eye and respiratory tract specimens. HEK cells were superior to HDF and MK cells in terms of both speed of virus detection and sensitivity. PMID- 3038952 TI - Comparison between indirect immunofluorescence and microneutralization for detection of antibodies to polioviruses. AB - Indirect immunofluorescence and microneutralization methods for the detection of antibodies to poliovirus serotypes 1, 2, and 3 were compared. Of the 41 sera tested for poliovirus type 1 antibody, 40% were in complete agreement, 55% differed by one dilution, and 5% differed by two dilutions. For poliovirus type 2, 37 sera were tested; 56% completely agreed, and 44% differed by only one dilution. For poliovirus type 3, complete agreement occurred in 59% of 33 sera, while 41% differed by one dilution. No false-negative results were obtained. These findings suggest that indirect immunofluorescence for poliovirus types 1, 2, and 3 is as sensitive as the microneutralization method and could represent a less cumbersome alternative. PMID- 3038953 TI - Parallel adaptation of the rabbit renal cortical sodium/proton antiporter and sodium/bicarbonate cotransporter in metabolic acidosis and alkalosis. AB - Recent studies have shown that the bicarbonate reabsorptive capacity of the proximal tubule is increased in metabolic acidosis. For net bicarbonate reabsorption to be regulated, there may be changes in the rate of apical H+ secretion as well as in the basolateral base exit step. The present studies examined the rate of Na+/H+ exchange (acridine orange method) and Na+/HCO3 cotransport (22Na uptake) in apical and basolateral membranes prepared from the rabbit renal cortex by sucrose density gradient centrifugation. NH4Cl loading was used to produce acidosis (arterial pH, 7.27 +/- 0.03), and Cl-deficient diet with furosemide was used to produce alkalosis (arterial pH, 7.51 +/- 0.02). Maximal transport rate (Vmax) of Na+/H+ antiporter and Na+/HCO3 cotransporter were inversely related with plasma bicarbonate concentration from 6 to 39 mM. Furthermore, the maximal transport rates of both systems varied in parallel; when Vmax for the Na+/HCO3 cotransporter was plotted against Vmax for the Na+/H+ antiporter for each of the 24 groups of rabbits, the regression coefficient (r) was 0.648 (P less than 0.001). There was no effect of acidosis or alkalosis on affinity for Na+ of either transporter. We conclude that both apical and basolateral H+/HCO3 transporters adapt during acid-base disturbances, and that the maximal transport rates of both systems vary in parallel during such acid base perturbations. PMID- 3038955 TI - Purification of human erythroid colony-forming units and demonstration of specific binding of erythropoietin. AB - Morphological and biochemical studies of human colony-forming units-erythroid (CFU-E) have been hindered by their extreme rarity. Since burst-forming units erythroid (BFU-E) develop into CFU-E, we used normal human blood BFU-E to generate large numbers of highly purified CFU-E in vitro. Using density centrifugation, sheep erythrocyte rosetting, surface immunoglobulin-positive cell depletion, adherence to plastic, and negative panning with monoclonal antibodies, human blood BFU-E were purified from 0.017 to 0.368%, a 22-fold purification with a 43% yield. The panned cells were cultured in methylcellulose with recombinant erythropoietin (rEp) and conditioned medium for 9 d. These cells were then collected and CFU-E were further purified using adherence and density centrifugation. This yielded almost 10(7) erythroid colony forming cells with a purity of 70 +/- 18%. Analysis of these cells by light and electron microscopy showed 94% erythroid cells. The prominent cell was a primitive blast with high nuclear/cytoplasmic ratio, dispersed nuclear chromatin and a distinct large nucleolus. The relation between the number of erythroid colonies and the number of day 9 cells plated in plasma clots was a straight line through the origin with a maximum number of erythroid colonies at 1 U/ml of rEp and no erythroid colonies without rEp. Specific binding with 125I-rEp showed that 60% of the binding was inhibited by excess pure erythropoietin (Ep), but not by albumin, fetal calf serum, and a variety of growth factors or glycoproteins. By days 12-13 of cell culture, when the progenitor cells matured to late erythroblasts, specific binding markedly declined. In this study, human CFU-E have been isolated in sufficient purity to characterize the morphology of these rare cells and in sufficient numbers to measure specific binding of Ep. PMID- 3038954 TI - Analysis of prostaglandin E2 effect on T lymphocyte activation. Abrogation of prostaglandin E2 inhibitory effect by the tumor promotor 12.0 tetradecanoyl phorbol-13 acetate. AB - We have investigated the inhibitory potential of prostaglandin E2 (PGE2) with respect to intracellular messengers implicated in the signaling system of T lymphocyte activation pathway. Using the fluorescent indicator Quin 2, it is demonstrated that PGE2 inhibits the increase in cytosolic-free calcium concentration [Ca2+]i. Reconstitution of calcium mobilization in the presence of PGE2 by the calcium ionophore A23187 results in a partial restoration of both interleukin 2 (IL2) production and cell proliferation and has no effect on the inhibition of transferrin receptor expression. In contrast, the treatment of cell cultures with the tumor promotor 12.0 tetra decanoyl phorbol-13-acetate (TPA) abrogates the suppressor activity of PGE2. When T lymphocyte stimulation is provided by the combination of A23187 and TPA, the PGE2 inhibitory effect does not occur. These data also indicate that the down regulation of transferrin receptor by PGE2 is proximal to protein kinase C activation and is not associated with decreased expression of the functional IL2 receptor. PMID- 3038956 TI - Pharmacological inhibition of in vitro infectivity of human T lymphotropic virus type I. AB - Human T lymphotropic virus type I (HTLV-I) is an exogenous RNA tumor virus etiologically linked to adult T cell leukemia and related diseases. In this paper, we describe that two 2',3'-dideoxynucleoside analogues, erythro 3'-azido 2',3'-dideoxythymidine (also called azidothymidine) and 2',3'-dideoxycytidine can inhibit the infectivity of HTLV-I against helper/inducer T cells in vitro. Both 2',3'-dideoxynucleoside analogues inhibited the overgrowth of target T cells, which was a consequence of virally mediated transformation, when they were exposed to the virus and cultured with the compounds. A profound decrease in the expression of HTLV-I gag-proteins was also observed. Moreover, we observed that the amount of proviral DNA detected in cellular DNA from the target T cells was substantially reduced when the cells were protected by the compounds against the virus and that at certain concentrations of the compounds the synthesis of viral DNA was completely suppressed. These results may be of value in developing a new pharmacological strategy for preventing the replication and possibly blocking the transmission of HTLV-I and related retroviruses in human beings. PMID- 3038957 TI - Lipoprotein binding to cultured human hepatoma cells. AB - Binding of various 125I-lipoproteins to hepatic receptors was studied on cultured human hepatoma cells (Hep G2). Chylomicrons, isolated from a chylothorax, chylomicron remnants, hypertriglyceridemic very low-density lipoproteins, normotriglyceridemic very low-density lipoproteins (NTG-VLDL), their remnants, low-density lipoproteins (LDL), and HDL-E (an Apo E-rich high-density lipoprotein isolated from the plasma of a patient with primary biliary cirrhosis) were bound by high-affinity receptors. Chylomicron remnants and HDL-E were bound with the highest affinity. The results, obtained from competitive binding experiments, are consistent with the existence of two distinct receptors on Hep G2 cells: (a) a remnant receptor capable of high-affinity binding of triglyceride-rich lipoproteins and HDL-E, but not of Apo E free LDL, and (b) a LDL receptor capable of high-affinity binding of LDL, NTG-VLDL, and HDL-E. Specific binding of Apo E free LDL was completely abolished in the presence of 3 mM EDTA, indicating that binding to the LDL receptor is calcium dependent. Specific binding of chylomicron remnants was not inhibited by the presence of even 10 mM EDTA. Preincubation of the Hep G2 cells in lipoprotein-containing medium resulted in complete suppression of LDL receptors but did not affect the remnant receptors. Hep G2 cells seem to be a suitable model for the study of hepatic receptors for lipoprotein in man. PMID- 3038958 TI - Type IIB von Willebrand factor with normal sialic acid content induces platelet aggregation in the absence of ristocetin. Role of platelet activation, fibrinogen, and two distinct membrane receptors. AB - Three preparations of purified von Willebrand factor (vWF), obtained from unrelated patients affected by type IIB von Willebrand disease, were found to have normal sialic acid content (between 129 and 170 nmol/mg of vWF, as compared with 158 +/- 17 nmol/mg in four normal preparations) and to induce platelet aggregation in the presence of physiologic levels of divalent cations and without addition of ristocetin. A monoclonal antibody that blocks the vWF binding domain of the platelet glycoprotein (GP)Ib caused complete inhibition of IIB vWF-induced aggregation. In contrast, a monoclonal antibody that blocks the receptor for adhesive proteins on the platelet GPIIb/IIIa complex failed to inhibit the initial response of platelets to high concentrations of IIB vWF. Moreover, IIB vWF caused agglutination of formalin-fixed platelets that was blocked only by the anti-GPIb antibody, suggesting that the binding of vWF to GPIb, even in the absence of ristocetin, results in platelet-platelet interaction that is followed by exposure of the GPIIb/IIIa receptors for adhesive proteins. Endogenous ADP, normally active platelet metabolism and fibrinogen binding to GPIIb/IIIa were necessary for maximal and irreversible platelet aggregation. In the absence of fibrinogen, however, aggregation was mediated by vWF binding to GPIIb/IIIa. A 52/48-kD tryptic fragment containing the GPIb binding domain of normal vWF completely blocked the aggregation induced by all three IIB vWF preparations. The present study defines in detail the mechanisms involved in IIB vWF-induced platelet aggregation. Moreover, it establishes that the GPIb binding domain of normal and IIB vWF are closely related and that desialylation is not required for the direct interaction of IIB vWF with GPIb. PMID- 3038959 TI - Apolipoprotein E2-Christchurch (136 Arg----Ser). New variant of human apolipoprotein E in a patient with type III hyperlipoproteinemia. AB - The primary structure of apolipoprotein E (apo E) was investigated in seven type III hyperlipoproteinemic patients with the apo E-2/2 phenotype. Six of the patients had identical two-dimensional tryptic peptide maps. These differed from the normal apo E3 map by the altered mobility of a single peptide. Amino acid analysis and sequencing showed that apo E2 in these patients had a substitution of 158 Arg----Cys. The presence of this mutation in six of the seven type III patients confirms that this is the most common form of apo E2. The seventh type III patient had a unique map with a new peptide resulting from a substitution of 136 Arg----Ser. He was heterozygous for this and for the more common apo E2 (158 Arg----Cys) variant. His very low-density lipoprotein contained approximately five times more apo E2 (136 Arg----Ser) than apo E2 (158 Arg----Cys), as determined by cysteamine treatment and peptide mapping. This new apo E2 mutant thus appears to contribute significantly to the patient's hyperlipidemia. PMID- 3038960 TI - Coxsackievirus B 1-induced polymyositis. Lack of disease expression in nu/nu mice. AB - Chronic inflammatory myositis similar to human polymyositis occurs in mice after infection with a strain of Coxsackievirus B 1 (CVB 1). To investigate the role of T cells in the pathogenesis of this disorder, we compared disease expression in T cell-deficient athymic nude (nu/nu) mice and heterozygotes (nu/+) with normal T cell function. Acute infectious myositis occurred in nu/nu and nu/+ mice. Chronic (greater than 21 d postinfection) weakness and myositis, however, developed only in nu/+. Resistance to disease in nu/nu mice was not explained by insusceptibility to infection; the amount of virus lethal for 50% of mice and virus replication were comparable in both groups. Additionally, anti-CVB 1 antibody production was similar in both groups. Reconstitution of infected nu/nu mice with spleen cells from normal mice resulted in disease. These results demonstrate that chronic weakness after infection with this virus is not simply a sequela of acute myonecrosis and suggest that T cells play a pivotal role in the pathogenesis of chronic myositis. PMID- 3038961 TI - Inhibition of the activation of Hageman factor (factor XII) by complement subcomponent C1q. AB - Hageman factor (HF, Factor XII) is activated by glass, collagen, and ellagic acid, and initiates blood coagulation via the intrinsic pathway. C1q inhibits collagen-induced platelet aggregation and adherence of platelets to glass, effects attributable to the collagen-like region of C1q. We examined the actions of C1q on HF activation. Incubation of C1q with HF before addition of HF deficient plasma extended the activated partial thromboplastin time. Similarly, when glass tubes were coated with C1q before testing, the partial thromboplastin time of normal plasma was increased. C1q reduced the activation of HF by ellagic acid, as measured by the release of p-nitroaniline from the synthetic substrate H D-prolyl-L-phenylalanyl-L-arginine-p-nitroanilide dihydrochloride, an effect inhibited by monoclonal anti-human C1q murine IgG and by digestion of C1q by collagenase. Thus, C1q inhibits activation of HF in vitro in clot-promoting and amidolytic assays and suggests a regulatory mechanism for the inhibition of coagulation. PMID- 3038963 TI - Phorbol myristate acetate, dioctanoylglycerol, and phosphatidic acid inhibit the hydroosmotic effect of vasopressin on rabbit cortical collecting tubule. AB - We explored the role for protein kinase C (PKC) in modulating vasopressin (AVP) stimulated hydraulic conductivity (Lp) in rabbit cortical collecting tubule (CCT) perfused in vitro at 37 degrees C. In control studies, 10 microU/ml AVP increased Lp (mean +/- SE, X 10(-7) centimeters/atmosphere per second) from 4.4 +/- 0.9 to 166.0 +/- 10.4. Pretreatment with dioctanoylglycerol (DiC8) suppressed AVP stimulated peak Lp (peak Lp, 21.9 +/- 3.1). Pretreatment with 10(-9) and 10(-7) M 4 beta-phorbol 12 beta-myristate 13 alpha-acetate (PMA) also blocked the increase in Lp in a dose-dependent fashion (peak Lp, 59.3 +/- 7.5 and 18.6 +/- 4.8, respectively). Inactive phorbol ester, 4 alpha-phorbol 12 beta,13 alpha didecanoate (10(-7) M), had no effect. PMA also suppressed the increase in Lp induced by 10(-4) M 8-p-chlorophenylthio-cyclic AMP (CcAMP): peak Lp was 169.4 +/ 14.9 in control, 79.2 +/- 5.5 with 10(-9) M PMA, and 25.7 +/- 2.9 with 10(-7) M PMA. Furthermore, when 10(-7) M PMA was added to the bath 10 min after exposure to AVP, the Lp response to AVP was blocked. Peak Lp was 52.4 +/- 9.6 with PMA vs. 165.1 +/- 10.0 in control. Phosphatidic acid (PA), which is thought to stimulate phosphatidylinositol (PI) turnover, produced similar inhibitory effects on AVP as well as CcAMP-stimulated Lp: PA suppressed 10-microU/ml AVP-induced peak Lp from a control value of 159.6 +/- 7.9 to 88.9 +/- 15.8, and 10(-4) M CcAMP induced peak Lp from 169.4 +/- 14.9 to 95.5 +/- 7.7. We conclude that PMA, at concentrations known to specifically activate PKC, suppresses the hydroosmotic effect of AVP on CCT; This suppression is primarily a post-cAMP event; Inhibition of AVP-stimulated Lp by DiC8 and PA also suggests an inhibitory role for the PKC system; The ability of pre- and post-AVP administration of PMA to blunt the AVP response suggests that agents that act through modulation of PI turnover in CCT may regulate the hydroosmotic effect of AVP. PMID- 3038962 TI - Stimulated mobilization of monocyte Mac-1 and p150,95 adhesion proteins from an intracellular vesicular compartment to the cell surface. AB - Monocytes were stimulated to increase their cell surface quantity of leukocyte adhesion proteins p150,95 and Mac-1 by the chemoattractant formyl-methionyl leucyl-phenylalanine, or other mediators such as platelet-derived growth factor, tumor necrosis factor, C5a, and leukotriene B4. Dose-response curves indicated variations in the sensitivity of monocytes and granulocytes to these mediators. These increases were independent of protein synthesis and half-maximal at 2 min. Human alveolar and murine peritoneal macrophages, cells that had previously diapedised, could not be induced to upregulate Mac-1 or p150,95. Detergent permeabilization studies in monocytes indicated that these proteins were stored in internal latent pools, which were reduced upon stimulation. Electron microscopy utilizing rabbit antiserum against p150,95 revealed these proteins on the plasma membrane, and in intracellular vesicles and peroxidase negative granules. Together with other functional studies, these findings suggest that the mobilization of Mac-1 and p150,95 from an intracellular compartment to the plasma membrane regulates the monocyte's ability to adhere and diapedese. PMID- 3038964 TI - Colorectal goblet cell mucins in familial adenomatous polyposis. AB - Ninety two tissue blocks from the left colon and 52 from the right colon were obtained from 112 patients with familial adenomatous polyposis (FAP). Tissues from 137 patients with other conditions served as controls. Within the main study a smaller investigation was performed to compare sections from the left and right colon in the same subject. Several well known histochemical techniques were used to investigate possible changes in sulphation, sialic acid structure (loss of O acetyl substituents), and changes in the ratio of sialic acid to neutral sugars. In patients with FAP, as in controls, there was increased expression of periodic acid Schiff positive mucin and fucose in the right colon. The only difference between patients with FAP and controls was the indirect demonstration of less neutral mucin in the right colon in FAP, but this did not seem to affect neutral sugars binding to UEA-1, PNL, or HPA. As in the general population, a small proportion of patients with FAP showed a lack of O-acetyl substituted sialic acid. Sialic acid heterogeneity probably has a genetic basis, but this is not associated with the genetic defect underlying FAP. PMID- 3038965 TI - Cytochemical profile of megakaryoblastic leukaemia: a study with cytochemical methods, monoclonal antibodies, and ultrastructural cytochemistry. AB - A cytochemical study using: Sudan black B; alpha-naphthyl acetate (ANAE) staining; estimation of alpha-naphthyl butyrate (ANBE) esterase activity; acid phosphatase activity; and 5' nucleotidase activity was carried out in 15 cases of megakaryoblastic leukaemia. These included cases of M7 acute myeloid leukaemia and blast crises of chronic granulocytic leukaemia. The megakaryoblastic nature of the blasts was first established using two monoclonal antibodies against platelet glycoproteins, and by estimating the platelet/peroxidase reaction at ultrastructural level. Our findings suggest that megakaryoblasts have a typical cytochemical profile comprising positive ANAE staining and acid phosphatase activity with a predominant localisation in the Golgi zone and negative or weak ANBE activity. A similar positive cytochemical pattern was also found in five cases of erythroleukaemia (M6). The specificity of the 5'nucleotidase activity for megakaryoblasts was not confirmed. In most cases of megakaryoblastic leukaemia there was no 5'nucleotidase activity only two cases showed positive reactions--reactions were positive in several cases of myeloblastic and lymphoblastic leukaemia. We suggest that cytochemical methods may be useful in diagnosing M6 and M7 acute leukaemia because less than 40% of leukaemic cells react with specific monoclonal antibodies. PMID- 3038966 TI - Routine full blood counts as indicators of acute viral infections. AB - Over a period of three weeks about 9000 full blood counts were analysed on the Technicon H6000 automated haematology machine. From these, 62 patients were identified who had abnormally high numbers of large unstained white cells; these patients were followed up for evidence of viral infection. Seventeen were either lost to follow up or in chronic renal failure; of the remaining 45 patients, 40 had viral infections, 26 of which were due to Epstein-Barr virus. In the presence of a raised number of large unstained white cells, an IgM test for Epstein-Barr virus is recommended, followed by routine serology when necessary. PMID- 3038967 TI - Rapid diagnosis of genital herpes by detecting cells infected with virus in smears with fluorescent monoclonal antibodies. PMID- 3038968 TI - Development of brainstem and cerebellar projections to the diencephalon with notes on thalamocortical projections: studies in the North American opossum. AB - The North American opossum is born in a very immature state, 12 days after conception, and climbs into an external pouch where it remains attached to a nipple for an extended period of time. We have taken advantage of the opossum's embryology to study the development of brainstem and cerebellar projections to the diencephalon as well as the timing of diencephalic projections to somatosensory motor areas of neocortex. The techniques employed included immunocytochemistry for serotonin, the retrograde and orthograde transport of wheat germ agglutinin conjugated to horseradish peroxidase, and the selective impregnation of degenerating axons. Our results suggest that serotoninergic axons, presumably from the dorsal raphe and superior central nuclei, are present in the diencephalon at birth. Axons from the bulbar reticular formation, the vestibular complex, the trigeminal sensory nuclei, and the dorsal column nuclei reach at least mesencephalic (and probably diencephalic) levels by postnatal day (PND) 3, whereas those from the cerebellar nuclei may not grow into comparable levels until PND 5. The dorsal column and cerebellar nuclei innervate the ventral nuclei of the thalamus by estimated postnatal day (EPND) 17 and all of the diencephalic nuclei supplied in the adult animal by EPND 26. Diencephalic axons enter ventrolateral (face) areas of presumptive somatosensory motor cortex by PND 12, but do not reach dorsomedial (limb) regions until EPND 21. At both ages, diencephalic axons are limited to the cortical subplate and marginal zone; they do not innervate an identifiable internal granular layer until considerably later. Our results suggest that axons from the brainstem and cerebellum grow into the diencephalon early in development, but that they do not influence the cerebral cortex until relatively late. When the results of the present study are compared with those reported previously on the development of ascending spinal (Martin et al., '83) and corticofugal (Martin et al., '80; Cabana and Martin, '85b,c) projections, it appears that specific components of major somatosensory and motor circuits develop according to different timetables. PMID- 3038969 TI - Spinal cord projections from hindlimb muscle nerves in the rat studied by transganglionic transport of horseradish peroxidase, wheat germ agglutinin conjugated horseradish peroxidase, or horseradish peroxidase with dimethylsulfoxide. AB - The spinal cord projections of four different groups of hindlimb muscle nerve branches--the medial and lateral gastrocnemius nerves, muscle branches of the deep peroneal nerve, muscle branches of the femoral nerve, and a nerve to the hamstring muscles--were studied with transganglionic transport of horseradish peroxidase (HRP) in the rat. The influence of varying the postoperative survival (3, 6, and 10 days) and of using wheat germ agglutinin-HRP conjugate (WGA-HRP), or HRP with dimethylsulfoxide (DMSO) instead of free HRP was studied for the gastrocnemius nerves. After 3 days' survival following application of HRP to the gastrocnemius nerves, fine granular labeling was found mainly in lamina V in L4 5, and coarse granular labeling was found in Clarke's column as far caudally as L2, and in laminae VI and VII predominantly in Th12-L2. After 6 or 10 days' survival, the fine labeling in lamina V was sparse or absent, whereas the coarse labeling appeared to remain or to be only slightly reduced in Clarke's column and in laminae VI and VII. No labeling suggestive of terminals was observed in laminae I-III from the gastrocnemius nerves. Except for sparse labeling in lamina I in some of the cases and some minor differences rostrocaudally, the spinal distribution of labeling was similar to that from the other nerves investigated. The distribution of labeling obtained after application of WGA-HRP or HRP with DMSO to the gastrocnemius nerves was very similar to that obtained with free HRP after 3 days' survival. The results indicate that the spinal cord projections of hindlimb muscle nerves in the rat distribute mainly in the deep part of the dorsal horn and in the intermediate zone. Furthermore, the lack of labeling suggestive of terminals in laminae I-III from the gastrocnemius nerves suggests, in conflict with earlier findings in the cat, that primary afferent fibers from muscles do not necessarily terminate in these laminae in the rat. The results suggest, furthermore, that fine granular labeling found in lamina V represents fine-calibered afferent fibers. Finally, the similar spinal projection patterns of the different muscle nerves investigated suggest either a less developed or an essentially different somatotopic organization for muscle afferents compared to cutaneous afferents, as revealed in earlier studies. PMID- 3038970 TI - Three primary neoplasms in a goat: hepatocellular carcinoma, phaeochromocytoma and leiomyoma. AB - A ten-year-old male nubian goat with no previous medical history was found recumbent. Clinical examination revealed an underweight, dehydrated, very depressed and unresponsive animal. At necropsy, the liver contained multiple, well-demarcated hepatocellular carcinomas. The right adrenal gland contained a solitary phaeochromocytoma. Multiple leiomyomas were present within the urinary bladder. The three primary neoplasms reported in this goat have been reported separately in other species; their simultaneous occurrence in one animal is uncommon. PMID- 3038972 TI - Cytoplasmic crystalline inclusions in hepatocytes of a steer with shipping fever. AB - Needle-shaped crystalline inclusion bodies were found in vacuolated liver cells of a steer affected with pneumonic pasteurellosis. The inclusions were eosinophilic and refractive, and ranged in size from 3 to 6 microns long and from 150 to 250 nm wide. Histochemical examination revealed that they contained lipid and protein. Electron microscopic examination showed finely fibrillar or granular material. Cross-striations with a periodicity of 45 nm were seen in longitudinal section of the inclusions. The possible origin and pathogenic significance of the inclusions are discussed. PMID- 3038971 TI - Pathological changes in HPCD pigs with prednisolone induced recrudescence of pseudorabies virus. AB - In HPCD pigs inoculated with PRV, latent PRV could be reactivated in-vivo by the administration of large doses of prednisolone 3 months after the primary infection. In two pigs, virus shedding was without clinical signs of disease, whereas depression of circulating lymphocytes was prominent. Reactivation of PRV was also demonstrated by cultivation of the brain cortex on the 7th day and the mandibular lymph node on the 9th day after the prednisolone began treatment. Coincident with the virus isolation, characteristic lesions were observed in 2 pigs in the central nervous tissues and mandibular lymph nodes and these were composed of cell necrosis and eosinophilic intranuclear inclusion bodies. Cells containing the intranuclear inclusion bodies had immature and mature PRV particles. Results of the present study with HPCD pigs indicated that the lesions in the brain and lymph node accompanied by eosinophilic intranuclear inclusion bodies were pathogonomonic lesions induced by reactivation of PRV. PMID- 3038973 TI - Synovial sarcoma of the jaw in a dog. AB - A case of synovial sarcoma of the jaw with pulmonary metastasis is described in a dog. It appears to be a rare or underdiagnosed neoplasm in animals and not previously reported in the jaw. Its diagnostic microscopic features are the biphasic cellular pattern and cleft formations. It may otherwise resemble haemangiopericytoma, malignant fibrous histiocytoma, reticulum cell sarcoma, fibrosarcoma, or giant-cell tumour of soft tissue. PMID- 3038974 TI - Primary cutaneous adenoid cystic carcinoma. AB - Primary adenoid cystic carcinoma of the skin is rare. Ten cases from the files of the Armed Forces Institute of Pathology were studied. Seven of the 10 cases recurred after initial surgery. The interval between surgery and recurrence ranged from 4 months to 20 years. In one case there was metastasis to lung and pleura. Histologically, cutaneous adenoid cystic carcinoma is indistinguishable from adenoid cystic carcinoma of the salivary gland. The diagnostic histologic features and the differentiation of adenoid cystic carcinoma from other cutaneous neoplasms are discussed. PMID- 3038975 TI - HSV-2 replication sites, monocyte and lymphocytic cell infection and virion phagocytosis by neutrophils, in vesicular lesions on penile skin. Electronoptical studies of a biopsy. AB - From a heterosexual male with recurrent genital herpes simplex virus (HSV-2) infection, a fresh intraepidermal vesicle on the penile skin was excised by punch biopsy, fixed and processed for electron microscopy. Differing locations and appearances of capsids and virions were studied to elucidate true host or destroyer cells. HSV-2 propagation and virion formation occurred predominantly in multi- or mononucleate spinosum cells situated at the base of the vesicle. However, some of the monocytes, young histiocytes and lymphocytic cells floating in the vesicle fluid were also involved. They harbored a small number of intranuclear capsids, designating the cells as viral (capsid) carriers. Infrequently encountered free virions in the vesicle fluid were invariably seen near neutrophils. All neutrophil granulocytes examined lacked intranuclear capsids. In contrast, distinct evidence of phagocytosis of virions and some capsids by neutrophils was found in the vesicle fluid near apical portions of spinosum cells packed with virions, or in neutrophils located between virion loaded spinosum cells in the base lining of the vesicle. In the cytoplasm of neutrophils, single and lysosome-enclosed clusters of virions were noted. Myelin figures and vacuolation of lysosomes in free-floating neutrophils were suggestive of virion distintegration. Viral propagation and abundant virion formation, beside neutrophil and lymphocyte attack, eventually lead to spinosum cell destruction. The minimal cytopathic effects (CPE) observed in involved monocytes and lymphocytic cells floating in the vesicle fluid suggest that these cells might function as vehicles for HSV-2 (capsid) transport to the exterior or interior. PMID- 3038976 TI - Effects of bicarbonate-based dental powder, fluoride, and saccharin on dental caries and on Streptococcus sobrinus recoveries in rats. AB - The effects of NaHCO3-based dental powder containing NaF and sodium saccharin on dental caries and Streptococcus sobrinus recoveries in rats were studied. Weanling specific-pathogen-free Osborne-Mendel (SPFOM) rats were inoculated with S. sobrinus 6715-13WT. One of six infected groups was topically treated with either demineralized water (DW), a dental powder suspended such that there was 1 part solid per 2 parts DW, 0.073% NaF, 0.5% Na-saccharin (Nas), or a combination of NaF and Nas at the same concentrations. NaF-supplemented DW (at 10 ppm F-) was provided to the 6th group of infected rats as a positive treatment control, but this group was otherwise untreated. A seventh but uninfected group was topically treated with DW. All topical treatments were given once for one min daily per rat, for five days per week. Animals' teeth were swabbed for recovery of 6715 13WT and total recoverable flora. At 37 days after start of treatment, S. sobrinus recoveries were lower only for those rats topically treated with the dental powder (p less than 0.05) by comparison with recoveries from the infected, topical DW-treated group. Caries scores, however, were 42% lower for the groups receiving the dental powder (p less than 0.005), 30% lower for those treated with the combined NaF-Nas (p less than 0.005), and 47% lower for the NaF-supplemented drinking water group (p less than 0.005). The dental powder effects, like those for the combined NaF-Nas and NaF drinking water, were evident on both smooth and fissure tooth surfaces. Both the 10 ppm F- drinking water and the dental powder significantly (p less than 0.005) reduced fissure caries scores below the level elicited by the indigenous mutans-free flora in the DW-treated uninfected rats; however, these reduction were not significantly different from one another. Thus, the 10 ppm F- drinking water and the dental powder equally inhibited not only the S. sobrinus-attributable component of caries but probably also the component of caries attributable to the indigenous oral flora. PMID- 3038977 TI - Selection for Streptococcus mutans with an altered xylitol transport capacity in chronic xylitol consumers. AB - The effect of long-term consumption of refined xylitol on the natural populations of S. mutans in the human oral cavity has been investigated. Fifty-four S. mutans strains were isolated from adults and children who had been consuming commercial food products containing xylitol for a period of from 1 1/2 to 10 years. Twenty isolates were also obtained from control subjects who had never consumed xylitol containing commercial food products. The inhibitory effect of xylitol on the isolated strains was determined by monitoring growth on glucose in the presence or absence of xylitol. This was used to define the sensitivity of each isolate to xylitol. Phosphoenolpyruvate:sugar phosphotransferase (PEP-PTS) activities were measured by means of the soluble and membrane fractions prepared from strains from both study populations. It was found that 87% of the fresh isolates from xylitol consumers were xylitol-resistant (XR), compared with only 10% of the strains isolated from the control subjects. The XR strains had low constitutive fructose PTS activity and very low xylitol-phosphorylating capacity. The xylitol sensitive (XS) strains, however, had much higher levels of constitutive fructose PTS activity and phosphorylated xylitol 16 times more rapidly than did the XR strains. Evidence for the phosphorylation of xylitol by a fructose PEP-PTS in the XS strains was obtained. The growth inhibition by the intracellular accumulation of non-metabolizable toxic xylitol phosphate and its prevention by the presence of fructose are discussed. PMID- 3038978 TI - Multiple linear eccrine spiradenoma associated with multiple trichoepithelioma. PMID- 3038979 TI - Localization of angiotensin converting enzyme (ACE) in granulomas of sarcoidosis cutaneous lesions. PMID- 3038980 TI - Rabbit zosteriform eruption induced by intra-arterial inoculation of herpes simplex virus, type 1. PMID- 3038981 TI - Cutaneous invasion of mucinous adenocarcinoma of the appendix. PMID- 3038982 TI - Regeneration and degradation of basal lamina-associated anionic sites in vitro. PMID- 3038983 TI - The pineal hormone melatonin in panic disorder. AB - The nocturnal synthesis of the pineal hormone melatonin was examined from 8 p.m. to midnight in 11 patients with panic disorder and eight control subjects. Patient concentrations of melatonin were significantly lower than controls at 10 p.m. (P less than 0.05; Kruskal-Wallis one-way ANOVA) and midnight (P less than 0.01). At 9 p.m. and 11 p.m., patient concentrations were also lower than controls, but were not statistically significantly different (P = 0.09). From these data, it is postulated that some patients with panic disorder exhibit a generalised defect in sensitivity of beta-adrenergic receptors or of sympathetic transmission. PMID- 3038984 TI - Plasma immunoreactive beta-endorphin in dexamethasone suppressors and non suppressors of cortisol. AB - Immunoreactive plasma beta-endorphin level was assayed in 33 patients with major affective disorder and in 16 psychiatrically normal controls before and after dexamethasone (1 mg) administration at 23.00 h. There were 18 cortisol suppressors and 15 non-suppressors among the patient group. All controls suppressed cortisol. Plasma beta-endorphin before dexamethasone was significantly different between suppressors, non-suppressors and controls (P less than 0.05, ANOVA). Concentrations of beta-endorphin were 4.7 +/- 0.7, 3.1 +/- 0.3 and 2.8 +/ 0.3 pmol/l for non-suppressors, suppressors and controls respectively. Following dexamethasone, beta-endorphin concentrations were again significantly different between groups (P less than 0.005, ANOVA). Concentrations were 4.6 +/- 0.7, 2.3 +/- 0.2 and 1.6 +/- 0.2 pmol/l for non-suppressors, suppressors and controls respectively. The implications of these findings are discussed. PMID- 3038985 TI - Dieting and weight loss in volunteers increases the number of alpha 2 adrenoceptors and 5-HT receptors on blood platelets without effect on [3H]imipramine binding. AB - Platelet monoamine receptor binding was determined in normal male and female subjects before and at the end of a 2- to 3-week weight reducing diet (1200 kcal daily). Dieting was associated with an increase in platelet binding sites for both [3H]yohimbine and [125I]iodolysergic acid diethylamide (iodoLSD). The affinity at the platelet [125I]iodoLSD binding site was reduced. In contrast, [3H]imipramine binding was unchanged. These results have important implications for studies that employ platelet binding as a peripheral marker of neurotransmitter function in psychiatric illness. PMID- 3038986 TI - Painful oral mucosal ulcers in a patient with small cell carcinoma of the lung. AB - This case illustrates the development of multiple painful oral ulcers caused by methotrexate that was one of a combination of chemotherapeutic drugs administered for the treatment of small cell carcinoma of the lung. Although the oral mucositis is self-limiting and resolves when the drug dose is reduced or therapy is discontinued, severe pain and discomfort may cause physical debilitation. Moreover, the risk of secondary oral infections is high in patients undergoing such therapy, and if the appropriate treatment is not instituted, fatal systemic dissemination of the infection may occur. PMID- 3038987 TI - The effect of nedocromil sodium on nasal provocation with allergen. AB - We have investigated the effect of nedocromil sodium in preventing the symptoms of rhinitis occurring immediately after allergen provocation in 10 patients with allergic rhinitis. The study had a double-blind, placebo-controlled, crossover design. Nedocromil sodium (1% w/v) and placebo were administered intranasally from identical metered-dose inhalers 20 minutes before allergen provocation. Three variables were measured: nasal airway resistance, the production of secretion, and the number of sneezes. Nedocromil sodium was significantly more effective than placebo in preventing nasal obstruction (p less than 0.01), rhinorrhea (p less than 0.01), and sneezing (p less than 0.05), suggesting that this drug may be useful in the treatment of allergic rhinitis. PMID- 3038988 TI - An investigation of human immune response to perennial ryegrass (Lolium perenne) pollen cytochrome c (Lol p X). AB - Cytochrome c (Lol p X) was purified from the pollen of perennial ryegrass (Lolium perenne). It was used in a sensitive double-antibody radioimmunoassay to determine specific IgG and IgE antibody (Ab) levels in the sera of grass-allergic human subjects and the relationship between immune responsiveness and HLA type. A total of 139 subjects were studied, including 63 subjects treated by grass immunotherapy and 76 untreated subjects. The sera of only four subjects were found to contain anticytochrome c IgG Ab; there was no evidence of cytochrome c specific IgE Ab in any of these patients. Based on these data, we conclude that the prevalence of IgE and IgG Ab responses to this protein is low. Because of the small number of responders, it was not possible to demonstrate a significant association between any HLA-D specificity and immune responsiveness to ryegrass cytochrome c. PMID- 3038989 TI - Hypothalamo-pituitary-adrenal function in infantile spasms: effects of ACTH therapy. AB - The metyrapone test was used to study the hypothalamo-pituitary-adrenal function in ten children with infantile spasms, before and after ACTH treatment. The hypothalamo-pituitary-adrenal response was normal before ACTH treatment in almost all children. After ACTH, the responses of two children were suggestive of a diminished pituitary reserve; three were suggestive of decreased adrenal as well as decreased pituitary reserve, and one suggested either adrenal hyperplasia with normal pituitary reserve, or appropriate response to a developing medical stress. We suggest that, in children being treated with ACTH, the dosage of ACTH should be gradually tapered, AM cortisol levels should be monitored, and high-dose steroids should be included when treating medical stress. PMID- 3038990 TI - Identification of calmodulin-like activity in term human amnion: effect of calmodulin inhibitors on prostaglandin biosynthesis. AB - Human amnion prostaglandin E2 (PGE2) synthesis increases with the onset of labour, and this synthesis is Ca2+-dependent. To understand better the mechanism of Ca2+-stimulated PGE2 biosynthesis, studies were performed to identify the presence of the intracellular Ca2+-mediator, calmodulin, in human amnion and to examine its role in PGE2 synthesis. Calmodulin-like activity was identified by the ability of the microsomal and cytosolic fractions of the 105,000g centrifugation of amnion homogenate to stimulate cyclic AMP-dependent phosphodiesterase activity. Cytosolic fractions consistently stimulated phosphodiesterase activity more than microsomal fractions (P less than 0.001) in paired samples from term human amnions. This activity was calcium-dependent. The cytosolic and microsomal factors increased the Vmax but not the Km of phosphodiesterase. There were no differences in these parameters with the onset of labour. The distribution of calmodulin-like activity between microsomes and cytosol was similar to the distribution of calmodulin mass as determined by radioimmunoassay. Three structurally different inhibitors of calmodulin activity, calmidazolium, trifluoperazine and W7, were tested for their ability to inhibit cytosolic factor-stimulated phosphodiesterase activity and to inhibit PGE2 output from dispersed amnion cells obtained before the onset of labour at term (cesarean section cells) or after spontaneous labour and vaginal delivery (spontaneous labour cells). The 50% inhibitory concentrations of the calmodulin antagonists in the phosphodiesterase assay were: trifluoperazine (6.7 microM), calmidazolium (0.11 microM), and W7 (24 microM). Trifluoperazine inhibited both basal and calcium ionophore (A23187)-stimulated PGE2 output from cesarean section cells and spontaneous labour amnion cells. Calmidazolium inhibited basal PGE2 output in cesarean section cells and spontaneous labour cells, but had no effect on A23187 stimulated output. W7 inhibited only the ionophore-stimulated PGE2 output in cesarean section amnion cells. The rank order of inhibition of both phosphodiesterase activation and basal PGE2 output was: calmidazolium greater than trifluoperazine greater than W7. These results suggest that human amnion contains calmodulin and that its distribution, concentration and activity remain unchanged with the onset of labour. The data suggest, although not conclusively, that calmodulin may, in part, play a role in amnion cell PGE2 production. Further investigation of calmodulin effects upon specific enzymes in the PGE2 synthetic pathway will be necessary to elucidate a role for calmodulin in PGE2 production. PMID- 3038991 TI - The relationship between fetal breathing movements and prostaglandin E2 during ACTH-induced labour in sheep. AB - We measured fetal breathing movements and fetal carotid arterial prostaglandin E concentrations during adrenocorticotrophin-induced labour in 6 pregnant sheep and in 6 control animals starting at day 127. The 6 ACTH-treated animals went into labour on average 97 h after the onset of infusion and the incidence of fetal breathing movements diminished during the last 12h before the onset of labour. There was a significant negative relationship between the incidence of fetal breathing movements and fetal carotid arterial prostaglandin E concentrations (r = -0.88; P less than 0.001) in ACTH treated animals. These data suggest a role for prostaglandin E in the diminution of fetal breathing movements prior to the onset of labour. PMID- 3038992 TI - Acute oral captopril inhibits angiotensin converting enzyme activity in human cerebrospinal fluid. AB - Angiotensin converting enzyme (ACE) activity in human plasma and cerebrospinal fluid (CSF), was measured by a fluorimetric method, following an acute oral dose (75 mg) of captopril. A decrease of approximately 60% in enzyme activity was observed in CSF, suggesting that the drug penetrates the blood/brain barrier (BBB) in sufficient amounts to inhibit the ACE in CSF. The activity of CSF enkephalinase, an enzyme present in human brain and inhibited in vitro by an elevated concentration of captopril, was, however, unaffected by the acute drug administration. It is proposed that the antihypertensive effect of captopril in humans may be due, at least in part, to the inhibition of ACE contained within brain structures. PMID- 3038993 TI - Elevated vascular angiotensin converting enzyme in chronic two-kidney, one clip hypertension in the dog. AB - The possible role of the vascular angiotensin converting enzyme (ACE) in the development of two-kidney, one clip (2-K, 1C) hypertensive dogs was studied in different blood vessels. Vascular ACE activity per mg protein differed in a variety of blood vessels; the activity appeared to vary inversely with the outer diameter of arteries. The systemic blood pressure in mongrel dogs increased after partial occlusion of the left renal artery, and the hypertension lasted for 8 months. Plasma renin activity (PRA) was raised only for the first 4 weeks after the operation and then returned to the original level in the chronic stage of hypertension. Plasma ACE activity did not alter during the experimental period. In contrast, ACE activities in the jejunal, pulmonary and renal arteries, aorta, lung and cerebral cortex, significantly increased in the chronic hypertensive stage (8 months after occlusion). The production of angiotensin II (ANG II) from ANG I was significantly greater in isolated arteries from 8-month hypertensive dogs than in those from normotensive dogs when assessed by the contractile responses to ANG I and ANG II. These results indicate that acceleration by increased vascular ACE activity of the production of ANG II in the vascular wall may contribute to the maintenance of hypertension in the chronic stage of 2-K, 1C hypertensive dogs having normal PRA and plasma ACE activity. PMID- 3038994 TI - Activity of cyclic GMP-dependent protein kinase in aortae from spontaneously hypertensive rats. AB - It has been suggested that various agents induce relaxation of vascular smooth muscles through guanosine 3',5'-cyclic monophosphate (cGMP) and cGMP-dependent protein kinase (cGMP-PK). In this work, the activity of cGMP-PK was studied in the 30,000 g supernatant from aortae of 4, 6, 8 and 12-week-old spontaneously hypertensive (SHR) and age-matched normotensive Wistar-Kyoto (WKY) rats and also of 4 and 12-week-old normotensive Wistar (W) and Sprague Dawley (SD) rats. At 4 weeks of age, both basal and cGMP-stimulated activity were not different in SHR and WKY rats. Nevertheless, a greater basal activity was measured in W (+50%) and SD (+20%) rats than in SHR, while no difference was observed between stimulated activities. In contrast with observations in the three normotensive rat strains, cGMP-PK activity did not decrease in the aortae supernatant of SHR rats aged 4-12 weeks. This resulted in mean increases of 45 and 30% in the basal and the cGMP stimulated activity, respectively, in the 12-week-old SHR rats. The abnormal evolution of cGMP-PK activity in the hypertensive strain was already detectable at 4-6 weeks of age. In apparent agreement with observations on protein kinase activity, cGMP binding activity attributable to cGMP-PK was 25% greater in 12 week-old hypertensive rats compared with age-matched WKY rats. These results indicate that in aortae of SHR rats, control of cGMP-PK activity is abnormal early in life. PMID- 3038995 TI - High angiotensin converting enzyme (kininase II) activity in the cerebrospinal fluid of spontaneously hypertensive adult rats. AB - Angiotensin converting enzyme (ACE, Kininase II, E.C. 3.4.15.1) activity was measured in the cerebrospinal fluid of 4- and 16-week-old spontaneously hypertensive rats (SHR) and age-matched Wistar-Kyoto (WKY) normotensive controls. Adult SHR showed higher cerebrospinal fluid enzyme activity than normotensive age matched WKY (19.6 +/- 1 and 32.3 +/- 5 nmol/h per ml in WKY and SHR, respectively, P less than 0.025). Conversely, there were no significant differences in enzyme activity in the cerebrospinal fluid of young animals. Our results support the hypothesis of enhanced activity of the central angiotensin system during the established phase of spontaneous hypertension in rats. PMID- 3038997 TI - Granulocyte-macrophage colony-stimulating factor augments the primary antibody response by enhancing the function of antigen-presenting cells. AB - Purified, recombinant-derived murine granulocyte-monocyte colony-stimulating factor was found to enhance the primary in vitro immune response to SRBC by murine spleen cells. In determining the mechanism of this augmentation, it was found that only splenic adherent cells and neither resting nor activated T cells nor B cells expressed specific receptors for GM-CSF. When splenic adherent cells were pulsed briefly with GM-CSF before addition to macrophage-depleted cultures, they reconstituted the PFC response to a significantly greater degree than did control macrophages. Splenic adherent cells incubated overnight with SRBC plus GM CSF were also more efficient antigen-presenting cells than splenic adherent cells incubated with antigen alone. The mechanism of this enhanced antigen presentation was found to be due to a GM-CSF-dependent increase in the level of IL 1 secretion and Ia antigen expression. Consistent with these data was the finding that GM-CSF augmented IL 2 production by splenic T cells in response to suboptimal concentrations of Con A. Finally, the day 5 in vivo antibody response (as measured by serum titers) of mice immunized with a low dose of SRBC was enhanced by two daily inoculations of GM-CSF. Thus, the role that GM-CSF plays in augmenting immune responses may not be solely accounted for by its ability to cause the proliferation or differentiation of macrophages, but more than likely includes its ability to enhance the function of antigen-presenting macrophages. PMID- 3038996 TI - Protein kinase C activation blocks anti-IgM-mediated signaling BAL17 B lymphoma cells. AB - Short term pretreatment of the B lymphoma, BAL17, with phorbol 12-myristate, 13 acetate (PMA) blocks elevation in inositol trisphosphate (InsP3) and increases in intracellular free calcium concentration ([Ca2+]i) in response to anti-IgM. The inhibition of enhanced InsP3 level is detected at 30 sec after the addition of anti-IgM, the earliest point measured, and is reversed by 1-(5 isoquinolinesulfonyl)-2-methylpiperazine dihydrochloride, an inhibitor of protein kinase C (PKC). The blockade of increased [Ca2+]i by PMA is also observed at the earliest time examined (15 sec), is reversed by 1-(5-isoquinoline-sulfonyl)-2 methylpiperazine dihydrochloride, and is mimicked by dioctanoylglycerol, a physiologic activator of PKC. The enhanced production of inositol phosphates in response to NaF is also blocked in BAL17 cells pretreated with PMA. Extended treatment of BAL17 cells with PMA depletes cellular PKC. Such pretreatment with PMA enhances rather than inhibits increased InsP3 levels in response to anti-IgM and leads to more sustained elevations in [Ca2+]i than in normal BAL17 cells. These results lead us to conclude that PMA-blockade of the response of B cells to anti-IgM represents a disruption of the transmembrane signaling process (desensitization of the signaling pathway) as a result of a PKC-mediated phosphorylation event. PMID- 3038998 TI - Down regulation of the receptors for tumor necrosis factor by interleukin 1 and 4 beta-phorbol-12-myristate-13-acetate. AB - Binding of radiolabeled tumor necrosis factor (TNF) to cell surface receptors was markedly reduced in human foreskin fibroblasts and cells from SV-80 and HeLa cell lines subsequent to treatment with interleukin 1 (IL-1) or 4 beta-phorbol-12 myristate-13-acetate (PMA). The decrease in TNF binding was initiated within minutes of application of IL-1 or PMA and could not be blocked by cycloheximide, suggesting that it is independent of protein synthesis. Scatchard plot analysis of TNF binding to the SV-80 cells indicated that its decrease in response to IL-1 and PMA reflects a reduced amount of TNF receptors, with no change in their affinity. IL-1 and PMA together had an additive effect on TNF binding. Treatment with TNF did not result in decreased binding of IL-1 to its receptors nor did TNF and IL-1 compete directly for their respective receptors. Human U937 cells on which receptors for IL-1 were below detectable levels exhibited no decrease in TNF binding when treated with IL-1, but did so in response to PMA. In addition to a decrease in TNF receptors, cells treated with IL-1 or PMA exhibited a lesser vulnerability to the cytolytic effect of TNF. The two kinds of changes were not completely correlated. A particularly notable dissimilarity was evident when comparing the rate of their reversal: the TNF receptor level was fully recovered within a few hours of removal of IL-1 or of the water-soluble analogue of PMA, 4 beta-phorbol-12,13-dibutyrate, from pretreated SV-80 cells; yet at that time resistance to the cytotoxicity of TNF was still prominent. These findings indicate that IL-1 as well as tumor-promoting phorbol diesters can down regulate cellular response to TNF by inducing a decrease in the number of receptors for TNF, and apparently through some other effect(s) as well. PMID- 3038999 TI - Regulation of interleukin 2 synthesis by cAMP in human T cells. AB - T cell activation requires two initial signals that first lead to the expression of interleukin 2 (IL 2) receptors and the initiation of IL 2 synthesis and then to T cell proliferation. Jurkat T lymphoma cells have been shown to be a good model for studying IL 2 synthesis because these cells also require two signals for activation. The first signal can be provided by the lectin phytohaemagglutinin (PHA), and the second one by the phorbol ester, 12-o tetradecanoylphorbol 13-acetate (TPA). The regulation of IL 2 synthesis in Jurkat cells, however, is unclear, and the present study deals with the role of cAMP on IL 2 synthesis. In Jurkat cells, IL 2 synthesis appears to be highly regulated by the activity of adenylate cyclase. This was demonstrated by using different means to increase intracellular cAMP level, namely by using permeant cAMP analogs, using the activator of adenylate cyclase, forskolin, using the activator of the alpha subunit of the stimulatory GTP binding protein cholera toxin, and using inhibitors of phosphodiesterase. In addition, prostaglandins E1 and E2 were shown to bind specifically to Jurkat cells, to induce a rise in intracellular cAMP level, and to markedly decrease IL 2 synthesis. All together, these results suggest that in T lymphocytes, the prostaglandin E2 receptor is linked to adenylate cyclase through a GTP binding protein and regulates the production of IL 2 by controlling the intracellular cAMP level. PMID- 3039000 TI - Assessment of leukotriene B4 synthesis in human lymphocytes by using high performance liquid chromatography and radioimmunoassay methods. AB - Leukotrienes (LT), mainly LTB4, have been shown recently to affect several functions of human lymphocytes in vitro, and they are regarded as putative modulators of the immune response. Although it is recognized that human neutrophils, eosinophils, monocyte-macrophages, and mast cells can generate LTs, the synthesis of 5-lipoxygenase products by lymphocytes is still the subject of a controversy. Human peripheral blood mononuclear leukocytes, nylon wool-purified lymphocytes, CD4+, CD4- T cells, large granular lymphocytes, and various fractions of pure lymphocyte preparations obtained by counter flow centrifugal elutriation were stimulated for 10 min to 24 hr with ionophore A23187, phytohemagglutinin, concanavalin A, or lipopolysaccharide with or without exogenous arachidonic acid (AA); supernatants were analyzed by reverse-phase high performance liquid chromatography (HPLC) coupled with radioimmunoassay (RIA) methods for the presence of LTB4. Pure human lymphocyte preparations, which were shown to be free of monocytes, did not release any detectable amount of LTB4. Increasing percentage of contaminating monocytes was clearly paralleled by increasing amounts of LTB4. Murine thymocytes, interleukin 2-dependent CTLL2 cytotoxic lymphocytes, EL4 thymoma cells, and human Jurkatt cells were also found to be unable to generate detectable amounts of LTB4 after stimulation with ionophore A23187, phytohemagglutinin, phorbol myristate acetate, recombinant interleukin 1, or interleukin 2 with or without exogenous AA. The addition of increasing numbers of adherence-purified monocytes to Jurkatt cells was followed by increased synthesis of LTB4. In conclusion, the present study indicates that the synthesis of LTB4 by pure human lymphocyte preparations or some human and animal lymphoid cell lines is not detectable by combined HPLC-RIA methods in any of the conditions used. PMID- 3039002 TI - Monocyte interleukin 2 receptor gene expression and interleukin 2 augmentation of microbicidal activity. AB - Activation of human peripheral blood monocytes results in the expression of interleukin 2 (IL 2) receptors, which are absent on resting monocytes. In a population of purified monocytes, the appearance of IL 2 receptors occurs on the majority of cells in association with increased levels of HLA-DR. Lipopolysaccharide (LPS) induces maximum numbers of IL 2 receptors within 12 hr, whereas IFN-gamma requires 48 hr. We used cDNA encoding for the human IL 2 receptor to evaluate IL 2 receptor gene expression in resting and activated monocytes. Within 4 hr after LPS stimulation, IL 2 receptor mRNA species of 3500 and 1500 bases appear, reaching peak levels between 8 and 12 hr and declining thereafter. The LPS-activated monocyte IL 2 receptor protein is expressed on the cell surface within a few hours after the detection of IL 2 receptor mRNA. The addition of IL 2 to IL 2 receptor-positive monocytes augments their generation of reactive oxygen intermediates and their cytotoxic activity. Thus monocytes when activated undergo a series of morphologic, phenotypic, and functional changes, including the expression of IL 2 receptors, which may provide an important immunoregulatory pathway. PMID- 3039001 TI - Beta-chain gene rearrangements and V beta gene usage in DPw2-specific T cells. AB - The diversity of human T cell receptor beta-chain gene rearrangements and variable region gene usage in the T cell response to a single allogeneic class II HLA gene product has been investigated. Nine clones of cytotoxic T lymphocytes (8.2 to 8.10) that are specific for the class II specificity DPw2 were analyzed for their T cell receptor beta-chain gene rearrangements using a constant-region probe. A minimum of seven different clonotypes were present in this panel of clones. The beta-gene expressed by one clone, 8.9, was isolated and the variable (V), diversity (D), and joining (J) segments were sequenced. The sequence of the V beta 8.9 segment is identical to the V beta 14 sequence, and is joined to D beta 1.1 and J beta 1.1 segments. Northern analysis revealed that three of the eight clones analyzed, 8.5, 8.7, and 8.9, expressed a 1.3 kb transcript that hybridized with the V beta 8.9 probe, indicating that these clones were using the same V beta gene (these clones shared common DNA rearrangements). The remaining five clones did not express V beta 8.9. Southern analysis of DNA obtained from a DPw2-specific tertiary mixed lymphocyte reaction bulk culture from which the clones were derived showed a prominent rearrangement of the V beta 8.9 gene that was indistinguishable from those observed for clones 8.5, 8.7, and 8.9. This prominent rearrangement of V beta 8.9 was not observed in DNA obtained from normal peripheral blood lymphocytes. These results suggest that although the number of V beta genes which can contribute to a DPw2 specificity may be relatively large, only a limited number of clonotypes ultimately predominate in the response to certain class II HLA antigens. PMID- 3039003 TI - Induction of human interleukin-1 production by polymyxin B. AB - Lipopolysaccharide (LPS) is a well known interleukin-1 inducer. Polymyxin B sulphate (PMB) is a cyclic antibiotic which neutralizes different biological properties of LPS. Under the conditions used in our laboratory, we were able to show that PMB alone could stimulate human monocytes to produce and release interleukin-1 activity, and that PMB was unable to inhibit the production of interleukin-1 by human monocytes stimulated with LPS. On the contrary, a synergistic effect was obtained which was not observed with another stimulant such as muramyl dipeptide. PMID- 3039004 TI - A colorimetric assay for the determination of 5'-nucleotidase activity. AB - A rapid, simple, quantitative and sensitive assay for the determination of 5' nucleotidase has been developed. The method can be applied to both soluble and membrane bound forms of the leukocyte enzyme. Enzyme activity is determined by colorimetric estimation of NH3 released from adenosine, the product of 5' nucleotidase activity in the presence of adenosine deaminase. The assay may be performed in microtitre plates and read with an automatic multiscan spectrophotometer. Thus it can be applied to a large number of samples for routine medical and research purposes. PMID- 3039005 TI - Human monoclonal antibodies against blood group antigens. Preparation of a series of stable EBV immortalized B clones producing high levels of antibody of different isotypes and specificities. AB - The EBV immortalization technique was used to produce stable clones, from B lymphocytes, secreting human monoclonal antibodies to Rh(D), Rh(G), Rh(c), Rh(E), Kell, A and A1 blood group antigens. These clones were obtained from peripheral blood lymphocytes of hyperimmunized plasmapheresis donors or from spleen lymphocytes of immunized patients. Mean levels of antibody concentration varied between 4 and 50 micrograms/ml. The antibodies obtained were of IgG1, IgG2, IgM or IgA class. Most of the clones have been stable for growth and antibody production during long periods of continuous culture, extending upto 4 years. Hybridization of two clones was effected with the human lymphoblastoid cell line KR-4 and with the mouse myeloma X63-Ag8.653, but did not result in any marked improvement of clone characteristics. One of the anti-Rh(D)-producing EBV transformed clones was used to produce an anti-Rh(D) typing reagent which has proved satisfactory for 2 years in routine blood typing in several laboratories. PMID- 3039007 TI - Tumours and tumour like conditions of testis. PMID- 3039006 TI - Selective removal of antigen-complexed IgG from cat plasma by adsorption onto a protein A-silica matrix. AB - The binding of normal cat IgG, heat-aggregated cat IgG and specific immune complexes (IC) containing cat IgG to a silica matrix containing covalently bound Staphylococcus aureus protein A was evaluated. The amounts of serum relative to protein A-silica, the flow rates and the perfusion times were representative of those existing when protein A-silica columns are used for therapeutic extracorporeal immunoadsorption of IgG and IC from humans and animals. When cat IgG was present in a large excess, approximately one molecule was bound to the matrix per molecule of solid-phase protein A with a KA of 1.5 X 10(6) 1/mol. Aggregated and immune complexed IgG bound to the matrix with relatively higher affinity. IC prepared in vitro between the purified envelope glycoprotein of the feline leukemia virus (FeLV gp70) and affinity-purified cat antibodies bound to the matrix even though normal IgG was present in greater than 10,000-fold excess. Once bound, IC were not eluted from columns upon further perfusion with normal serum. However, bound IgG was eluted from columns by further perfusion of normal serum or IC. IC were at least five-fold more efficient than normal IgG in exerting this effect. The results suggest that protein A-silica columns can be used for preferential removal of IC from plasma in a clinical or experimental setting. PMID- 3039008 TI - Drug-induced ventricular tachycardia in kala-azar. PMID- 3039009 TI - Antibody response to herpes simplex virus glycoprotein D: effects of acyclovir and relation to recurrence. AB - We developed an enzyme-linked immunosorbent assay (ELISA) for measuring the antibody response to herpes simplex virus (HSV) glycoprotein D (gD). The ELISA was specific and more sensitive than immunoblotting techniques. Antibody to HSV gD was detected in 10 of 12 sera (geometric mean titer, 36.3) obtained from HSV-2 infected guinea pigs 14 days after intravaginal inoculation, and the titer increased to greater than or equal to 5,120 on day 60. Therapy with acyclovir delayed and diminished the antibody response to gD, although by day 60 the titers of antibody to gD in acyclovir-treated animals were not significantly different from those in controls. An increased titer of antibody to gD seemed to be associated with a reduced number of recurrent episodes and to a reduced number of days with recurrent lesions. PMID- 3039010 TI - Antibodies to the three major glycoproteins of varicella-zoster virus: search for the relevant host immune response. AB - Clinical manifestations of chickenpox have occurred in individuals known to have seroconverted after vaccination against varicella. To determine whether these "breakthroughs" might be due to the absence of specific antibodies, we tested sera from vaccinees before or at the time of exposure to varicella for antibodies to the three major glycoproteins of varicella-zoster virus (VZV). Protection could not be correlated with the presence or level of any of these antibodies. Levels of antibodies to glycoproteins before onset of zoster were similar to those in sera of individuals who had had varicella previously and did not develop varicella after household exposure. Thus, protection against infections with VZV cannot be explained by the presence of specific antibodies to glycoproteins. PMID- 3039011 TI - Aerosol transmission of rhinovirus colds. AB - Rhinovirus infections may spread by aerosol, direct contact, or indirect contact involving environmental objects. We examined aerosol and indirect contact in transmission of rhinovirus type 16 colds between laboratory-infected men (donors) and susceptible men (recipients) who played cards together for 12 hr. In three experiments the infection rate of restrained recipients (10 [56%] of 18), who could not touch their faces and could only have been infected by aerosols, and that of unrestrained recipients (12[67%] of 18), who could have been infected by aerosol, by direct contact, or by indirect fomite contact, was not significantly different (chi 2 = 0.468, P = .494). In a fourth experiment, transmission via fomites heavily used for 12 hr by eight donors was the only possible route of spread, and no transmissions occurred among 12 recipients (P less than .001 by two-tailed Fisher's exact test). These results suggest that contrary to current opinion, rhinovirus transmission, at least in adults, occurs chiefly by the aerosol route. PMID- 3039012 TI - In vivo Streptococcus pyogenes C5a peptidase activity: analysis using transposon- and nitrosoguanidine-induced mutants. AB - The streptococcal C5a peptidase removes a six-amino-acid fragment from human C5a and thereby inactivates this chemotaxin. We used transposon and chemical mutagenesis to generate mutants of Streptococcus pyogenes that did not produce C5a peptidase. These mutants showed no alteration in expression of capsule, M protein, streptolysins O and S, or pyrogenic exotoxin C. Serial passage of a peptidase-producing strain in vivo resulted in a 100-fold increase in production of C5a peptidase. The presence of C5a peptidase delayed the accumulation of polymorphonuclear leukocytes (PMNLs) in the peritoneal cavities of mice after intraperitoneal challenge. However, there was no difference in virulence (as evaluated by LD50) between strains that produced and those that lacked C5a peptidase. Although C5a peptidase is expressed on the cell surface, antibody to this enzyme did not opsonize streptococci for phagocytosis in vitro. These studies show that C5a peptidase alters the normal host inflammatory response by delaying the accumulation of PMNLs at the foci of streptococcal infection. PMID- 3039013 TI - Structure and expression in Escherichia coli of canine interferon-alpha genes. AB - Using a human interferon-alpha (IFN-alpha) cDNA probe, several recombinant phages containing type I IFN genes were isolated from a canine genomic library. One of these phages contains two complete CaIFN-alpha genes with identical coding sequences, and a second one a slightly different IFN-alpha gene. The IFN-alpha protein sequences contain six cysteine residues as well as two or three potential N-glycosylation sites. Expression of mature CaIFN-alpha 1 in E. coli results in antiviral activity on dog cells. Genomic analysis using an equine IFN-omega probe and DNA sequencing suggests the deletion of IFN-omega genes from canine genome. PMID- 3039014 TI - Potency stability of recombinant (Serine-17) human interferon-beta. AB - The antiviral activity of Escherichia coli-derived (Serine-17) human interferon beta, formulated with human serum albumin, is stable for 2 years when lyophilized and stored under refrigeration. This product shows an Arrhenius line fit for the stability of its activity when tested at multiple isothermal temperatures (25-80 degrees C). In both isothermal and nonisothermal elevated temperature studies, increasing the level of human serum albumin in the formulation results in increased thermal stability. PMID- 3039015 TI - Recombinant human interferon-alpha A treatment of an experimental cutaneous herpes simplex virus type 1 infection of guinea pigs. AB - Recombinant human interferon-alpha A (rIFN-alpha A) was evaluated for therapeutic efficacy against an experimental dorsal cutaneous herpes simplex virus type 1 (HSV-1) infection of guinea pigs. Human IFN has activity in this species. Animals were treated either systemically (i.m.) or topically with different formulations of rIFN-alpha A. Therapy was initiated 24 h before (-24 h), 30 min after (+0.5 h), or 24 h following (+24 h) virus inoculation, and treatment was continued for 3-5 days. Efficacy was evaluated on day 4 following infection. Treatment with rIFN-alpha A given systemically beginning at -24 h was effective, reducing lesion number, total lesion area, and lesion virus titer by 64%, 85%, and 98% respectively (p less than 0.001). Efficacy diminished with delay in initiation of i.m. therapy. Topical rIFN-alpha A formulations were generally ineffective, showing only a marginal effect with therapy initiated -24 h or +0.5 h and no effect when treatment was delayed to +24 h. In summary, rIFN-alpha A is effective against cutaneous HSV-1 infection in this model if therapy is initiated prophylactically and if drug delivery is assured by systemic injection. Topical application of rIFN-alpha A showed little therapeutic effect. The large molecular weight of IFN likely retards its percutaneous delivery. PMID- 3039017 TI - Preformed hydroxylapatite blocks for palatal grafting in orthognathic surgery. AB - Hydroxylapatite has been used as a bone substitute for ridge augmentation, ridge maintenance and periodontal defects for many years. This is a preliminary report of the use of hydroxylapatite blocks as a grafting material in midpalatal splits in orthognathic surgery. With the advent of hydroxylapatite blocks, the need for autogenous bone grafting will, in most cases, be obviated. PMID- 3039016 TI - Issues in drug management. Part 2. The use of receptor-selective agents as analgesics in the spinal cord: trends and possibilities. PMID- 3039018 TI - [Herpes zoster in patients with systemic lupus erythematosus]. PMID- 3039020 TI - [Papillomavirus infection of the vulva]. AB - The role of Papillomavirus (PV) in the development of external genital neoplasias has recently been documented. Fifty cases of condyloma acuminata, dystrophy, Bowenoid papulosis, carcinoma in situ and invasive carcinoma of the vulva were examined for the presence of PV structural antigens with an immunoperoxidase method on formalin-fixed, paraffin-embedded tissue. Using the avidin-biotin peroxidase complex (ABC) technique on vulvar tissue obtained from biopsy and vulvectomy specimens, PV structural antigens were identified in association with condyloma acuminata in 5 of 6 cases (83.3%), with hyperplastic dystrophy in 1 of 11 cases (9.1%), with atypia in 9 of 14 cases (64.3%), with Bowenoid papulosis in 2 of 2 cases (100%), with carcinoma in situ in 4 of 7 cases (57.1%) and with invasive carcinoma in three of 3 cases (100%), but in none of the examples of 7 lichen sclerosus cases. A PV-inducing morphologic change in koilocytosis was present in 30 instances of all the vulvar lesions. The results of this study, therefore, provide immunohistochemical evidence demonstrating the close relationship of papillomavirus to vulvar disease ranging from condyloma acuminata and moderate atypia to carcinoma in situ and invasive carcinoma. PMID- 3039019 TI - [Cholesterol metabolism in macrophages or macrophage-derived cells]. PMID- 3039021 TI - Ultrasonic evaluation of trophoblastic disease and the response to chemotherapy. AB - Ultrasound is an established method of confirming the presence of a hydatidiform mole in utero. However, in agreement with other investigations, we have often noted that the appearance of this entity is less specific than originally reported. To elucidate this, the ultrasonic patterns of 26 patients with trophoblastic disease and 27 with other entities were reviewed. Since there is a paucity of literature with regard to the response of choriocarcinoma to chemotherapy, determined by ultrasound, we simultaneously studied the relationship between findings of the ultrasonograms and the levels of hCG-beta subunit in sera. We noted variable ultrasonic features in trophoblastic disease, and the sonograms of the choriocarcinoma have occasionally been confused with those of hydatidiform mole. If special attention is directed to the thickness of the placenta-like echoes as well as the sonolucent areas within it, the diagnosis of partial hydatidiform mole may be feasible. We also noted a rough correlation between ultrasonic appearances and the hCG-beta-subunit value, determined during chemotherapy for choriocarcinoma. PMID- 3039022 TI - [A case of idiopathic Addison's disease with pregnancy]. PMID- 3039023 TI - [Postoperative neutron irradiation in patients with primary lung cancer]. PMID- 3039024 TI - [Changes in blood coagulation and fibrinolysis after transcatheter arterial chemoembolization in hepatocellular carcinoma]. PMID- 3039025 TI - [Partial deficiency of cytochrome c oxidase in skeletal muscles; two cousins presenting progressive external ophthalmoplegia and proximal weakness of the limbs]. PMID- 3039026 TI - [A case of hepatocellular carcinoma associated with polyphagia due to midbrain tumor embolism]. PMID- 3039027 TI - [Effects of dibutyryl cyclic AMP on the left ventricular filling dynamics]. PMID- 3039028 TI - [A case of glucagonoma presenting with epigastric and back pain as an initial manifestation]. PMID- 3039029 TI - [Apatite ceramics implant]. PMID- 3039030 TI - An assessment of inflammation in the reservoir after restorative proctocolectomy with ileoanal ileal reservoir. AB - The significance of inflammation of the mucosa of the ileal reservoir after restorative proctocolectomy is not known although in some cases it appears to be associated with symptoms when the condition has been referred to as pouchitis. This investigation has aimed to determined the prevalence of inflammation, to define pouchitis and to examine some factors which might be related to inflammation. Mucosal biopsies from the ileal reservoir were studied in 90 patients at up to 62 months after closure of the ileostomy. A histological grading system (0-6) was used to assess the severity of inflammation. Some degree of chronic and acute inflammation was found in 87% and 30% of cases respectively. The prevalence of a grade of 4 or more was 23% and 3.5%. There was a correlation between severity of chronic and acute inflammation. Severe histological acute inflammation (grade 4-6) was associated with sigmoidoscopic features of inflammation and with increased frequency of defaecation. Of 55 patients sigmoidoscoped by one clinician, 6 (11%) had pouchitis which was characterised by macroscopic inflammation of the reservoir, diarrhoea and a histological grade of 4 or more. The severity of chronic inflammation was not related to frequency of defaecation. Histological inflammation could not be correlated with the type of reservoir, residual volume after evacuation of a known volume of stool substitute introduced per anum into the reservoir or compliance of the reservoir. Acute inflammation was significantly more severe in patients with ulcerative colitis than in those with familial adenomatous polyposis. PMID- 3039031 TI - Adenosine: emerging role as an immunomodifying agent. PMID- 3039032 TI - Adenosine receptors on rabbit alveolar macrophages: binding characteristics and effects on cellular function. AB - Adenosine and its synthetic analogues are known to affect many leukocyte functions, in some cases by binding to specific cell surface receptors coupled to adenylate cyclase. In this study, adenosine receptors were demonstrated on normal rabbit alveolar macrophages by examining specific binding of tritiated 5-N ethylcarboxamide adenosine (NECA) to intact cells. Scatchard analysis suggested a single class of approximately 33,000 binding sites per cell and an estimated Kd of 0.46 mumol/L. Competitive inhibition of tritiated NECA binding was demonstrated for 2-chloroadenosine (2-CA; Ki = 3.68 mumol/L) and L phenylisopropyl adenosine (L-PIA; Ki greater than 100 mumol/L), a rank order of binding affinities indicative of an A2 receptor. Theophylline and isobutyl methylxanthine had Kis of 368 and 27.6 mumol/L, respectively. For functional correlation, NECA was found to be 10-fold more potent than L-PIA in stimulating an increase in intracellular cyclic adenosine monophosphate. In addition, macrophages were cultured for 24 hours with NECA, 2-CA, or L-PIA to determine whether these analogues modulated expression of either cell-associated procoagulant activity or elaboration of plasminogen activator. Procoagulant activity was suppressed by as much as 62% (P less than 0.05); the rank order of potency and blockade of the effect with theophylline suggest that suppression of procoagulant activity occurred primarily by stimulation of A2 receptors. By contrast, these analogues stimulated production and release of plasminogen activator by 30% (P less than 0.05), but this effect had none of the features of an A2-mediated mechanism. Macrophages were cotreated with nitrobenzylthioinosine (10 mumol/L) and adenosine deaminase (2 U/ml) to allow adenosine accumulation exclusively within the cell.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3039033 TI - Solitary thyroid nodule: radionuclide study at Songkhlanagarind Hospital. PMID- 3039034 TI - Activation of the opioid and nonopioid analgesic systems: evidence for a memory hypothesis and against the coulometric hypothesis. AB - It has been suggested that the magnitude and form of hypoalgesia elicited by an aversive event can be predicted from its coulometric product (Intensity X Duration). According to this hypothesis, small products elicit opioid hypoalgesia, and large products elicit nonopioid hypoalgesia. This suggests that increasing the duration of an aversive event should heighten the nonopioid hypoalgesia. Contrary to this prediction, in Experiment 1 I found that increasing the duration of a mild shock attenuated the nonopioid hypoalgesia. In Experiment 2 I tested another implication of the coulometric hypothesis, namely, that mild shocks that have the same coulometric product should elicit equivalent hypoalgesia. The results did not support this prediction. We discuss how these findings are consistent with an alternative theory, the "working memory hypothesis." According to this theory, the representation of an aversive event in working memory elicits hypoalgesia. In Experiment 3 a novel prediction of this theory was tested, namely, that displacing the representation of intense shock from working memory, by following the intense shock with a weak shock "distractor", should attenuate hypoalgesia. The results support this prediction. I conclude by discussing the relation of this work to other findings in the analgesia literature. PMID- 3039035 TI - Shock signals and the development of stress-induced analgesia. AB - We report five experiments in which we investigated the effects of "feedback signals" on the pattern of hypoalgesia produced by inescapable shocks. A 5-s lights-out stimulus coincident with shock termination had no effect on the naltrexone-insensitive (nonopioid) hypoalgesia, which occurred after 10 inescapable shocks, but completely blocked the naltrexone-sensitive (opioid) hypoalgesia, which followed 100 inescapable shocks. The stimulus prevented the development of the opioid hypoalgesia rather than merely masking its measurement. This effect did not depend on the use of lights-out as the stimulus but did depend on the temporal relation between the stimulus and shock. Stimuli immediately preceding or simultaneous with shock had no effect. Surprisingly, stimuli randomly related to shock also blocked the opioid hypoalgesia. Simultaneous measurement of both hypoalgesia and fear conditioned to contextual cues revealed that the level of fear did not predict the blockade of hypoalgesia. Different backward groups received different temporal gaps between shock termination and the signal. An interval between 2.5 s and 7.5 s eliminated the effect of the signal on fear, but 12.5-17.5 s were required to eliminate the effect of the signal on hypoalgesia. The opioid hypoalgesia blocking power of the random stimulus was entirely attributable to those stimuli occurring within 15 s of the termination of the preceding shock. The implications of these results for the explanation of stimulus feedback effects and for stress-induced analgesia are discussed. PMID- 3039036 TI - Short-term changes in levels of cyclic AMP, adenylate cyclase, and phosphodiesterase during the initiation of sperm motility in rainbow trout. AB - In order to clarify the role of the system that generates and degrades cyclic AMP during the initiation of motility of trout sperm, short-term changes in levels of intraspermatozoal cyclic AMP, adenylate cyclase, and phosphodiesterase were measured. Levels of cyclic AMP and the activity of adenylate cyclase increased and reached a maximum level 1 sec after transfer of sperm to K+-free medium, where they became motile, and then decreased rapidly. However, there were no changes in either parameter in sperm which remained immotile in K+-rich medium. In addition, an increase in the activity of phosphodiesterase was observed 4 sec later than the increase in levels of cyclic AMP and adenylate cyclase. These findings suggest that a very rapid change in the level of intracellular cyclic AMP occurs within 1 sec, at the moment of spawning, by the activation of adenylate cyclase and phosphodiesterase, and regulates the initiation of trout sperm motility. PMID- 3039038 TI - The common cold--my favourite infection. The eighteenth Majority Stephenson memorial lecture. PMID- 3039037 TI - ADP binding to myosin cross-bridges and its effect on the cross-bridge detachment rate constants. AB - We have studied the binding of adenosine diphosphate (ADP) to attached cross bridges in chemically skinned rabbit psoas muscle fibers and the effect of that binding on the cross-bridge detachment rate constants. Cross-bridges with ADP bound to the active site behave very similarly to cross-bridges without any nucleotide at the active site. First, fiber stiffness is the same as in rigor, which presumably implies that, as in rigor, all the cross-bridges are attached. Second, the cross-bridge detachment rate constants in the presence of ADP, measured from the rate of decay of the force induced by a small stretch, are, over a time scale of minutes, similar to those seen in rigor. Because ADP binding to the active site does not cause an increase in the cross-bridge detachment rate constants, whereas binding of nucleotide analogues such as adenyl-5'-yl imidodiphosphate (AMP-PNP) and pyrophosphate (PPi) do, it was possible, by using ADP as a competitive inhibitor of PPi or AMP-PNP, to measure the competitive inhibition constant and thereby the dissociation constant for ADP binding to attached cross-bridges. We found that adding 175 microM ADP to 4 mM PPi or 4 mM AMP-PNP produces as much of a decrease in the apparent cross-bridge detachment rate constants as reducing the analogue concentration from 4 to 1 mM. This suggests that ADP is binding to attached cross-bridges with a dissociation constant of approximately 60 microM. This value is quite similar to that reported for ADP binding to actomyosin subfragment-1 (acto-S1) in solution, which provides further support for the idea that nucleotides and nucleotide analogues seem to bind about as strongly to attached cross-bridges in fibers as to acto-S1 in solution (Johnson, R.E., and P. H. Adams. 1984. FEBS Letters. 174:11-14; Schoenberg, M., and E. Eisenberg. 1985. Biophysical Journal. 48:863-871; Biosca, J.A., L.E. Greene, and E. Eisenberg. 1986. Journal of Biological Chemistry. 261:9793-9800). PMID- 3039039 TI - Detection of Epstein-Barr virus strain variants in lymphoblastoid cell lines 'spontaneously' derived from patients with rheumatoid arthritis, infectious mononucleosis and normal controls. AB - 'Spontaneous' lymphoblastoid cell lines (LCL) were established from patients with either rheumatoid arthritis (RA) or infectious mononucleosis (IM) or from healthy donors. Differences in Epstein-Barr virus (EBV) strains were determined by measuring the mol. wt. and expression of viral antigens in each of the LCLs. In addition to the previously reported EBV nuclear antigens, the LCLs also contained EBV-induced antigens with mol. wt. of 48K and 58K which were present in all but two of the lines. One of the differences observed between each of the groups of cell lines was their ability to produce viral antigens. Early and late antigens were identified by immunoblotting in most of the RA lines, two of the normal lines but none of the cell lines from patients with IM. Many of the IM cell lines were also found to express multiple EBNA1 antigens. The results demonstrate that a variety of wild-type EBV strains exist. However, the similarities observed in a number of the lines suggest that the diversity of strains may be limited. PMID- 3039041 TI - Generation and properties of the glycoprotein E-related 32K/34K/35K and 55K/57K polypeptides encoded by herpes simplex virus type 1. AB - A hybridoma line was isolated which produced antibody reacting with polypeptides of apparent molecular weights 32,000, 34,000 and 35,000 (32K/34K/35K) and 55,000 and 57,000 (55K/57K). These were sulphated glycoproteins that have previously been found in the medium of herpes simplex virus type 1 (HSV-1)-infected cells. By tryptic peptide mapping and serological cross-reactions the polypeptides were shown to be related to HSV-1 glycoprotein E (gE-1) but they lacked the Fc binding function. The 32K/34K/35K and 55K/57K polypeptides were not found in the medium of HSV-1-infected cells incubated in serum-free medium. They could be generated in vitro from purified gE-1 in the presence of serum. It is likely that 32K/34K/35K and 55K/57K are derived from gE-1 by the action of serum proteases. PMID- 3039040 TI - Herpes simplex virus replication and protein synthesis in a human blood-derived cell line. AB - Herpes simplex virus (HSV) types 1 and 2 were shown to replicate in a newly described human cell line (Meg) derived from the peripheral blood of a healthy volunteer. The cell line has both megakaryocyte-like and B cell-like properties. Upon infection with HSV-1 or -2, at a m.o.i. between 0.5 and 5, unlike B and T cells, the Meg cells were growth-arrested and this was accompanied by cytopathic effects and virus replication. The HSV proteins and glycoproteins B and D (gB and gD) made in the blood-derived Meg cells were compared to the corresponding proteins made in the non-blood-derived cell lines, Vero (African green monkey kidney cell) and HEp-2 (human epidermoid carcinoma cell). The maximum level of HSV protein synthesis occurred earlier in the Meg cells than in the Vero and HEp 2 cells. The electrophoretic pattern of HSV-1 and -2 proteins made in the Meg cell line was similar to the corresponding proteins made in the Vero and HEp-2 cell lines; however, some qualitative and quantitative differences were evident. There were no apparent differences detected in the migration pattern of gB made in all three cell lines while significant differences were observed with the gD species. However, upon hydrolysis with Staphylococcus aureus V-8 protease of the monoclonal antibody-purified gB and gD, distinct differences were observed in the electrophoretic pattern of the generated peptide fragments of both gB and gD made in the three cell lines. The results demonstrate that a human blood cell can support HSV replication and that species-specific post-translational modification of gB and gD occurs in HSV-infected Vero cells as compared to HSV-infected human cells. PMID- 3039042 TI - Genome variations in herpes simplex virus type 2 strains isolated in Japan and Sweden. AB - One-hundred and twenty-three epidemiologically unrelated strains of herpes simplex virus type 2 (HSV-2) isolated in Japan and Sweden (68 Japanese and 55 Swedish isolates) were compared by analysis of their genomes using five restriction endonucleases: BamHI, KpnI, EcoRI, HindIII and Bg/II. Seven of the 93 restriction sites examined showed statistically significant variation between isolates from the two countries. However, HSV-2 isolates were less variable than the HSV-1 isolates previously analysed from the same countries. Using 12 restriction sites as markers, the HSV-2 isolates were classified into 41 cleavage patterns; 17 were specific for Japanese isolates and 15 were specific for Swedish isolates. Correlation coefficients between some sets of 12 markers were significant, but significant correlations between Japanese and Swedish isolates were distinct for each country. Both Japanese and Swedish isolates were assigned to three major patterns with no significant difference in incidence. In contrast, in two other major patterns, differences in incidence between the isolates were statistically significant. These results suggest that HSV-2 populations in geographically separated countries have distinct cleavage site distributions. PMID- 3039043 TI - The nucleotide sequence and genome organization of bovine papillomavirus type 4. AB - The nucleotide sequence of bovine papillomavirus type 4 (BPV-4) was determined. The viral genome is 7261 base pairs long. Several overlapping open reading frames (ORFs) have been identified both on the basis of amino acid comparison with other papillomaviruses and on their transcriptional pattern. Eight early ORFs (E1 to 8) were recognized, coding for DNA replication and cell transformation functions, and three late ORFs (L1 to 3), coding for structural proteins. Like the E5 ORF of human papillomavirus type 6 the E5 ORF of BPV-4 is discontinuous. Unlike other papillomaviruses, the non-coding region upstream of the early ORFs (ncr-1) is short (385 base pairs), but there is another non-coding region (ncr-2) of nearly 500 base pairs between the L2 and L1 ORFs. Most of the putative regulatory sites are located in the ncr-1, although potential controlling elements are also found in other parts of the genome. Polyadenylation sites are present at the 3' end of both the early and the late transcription units. Comparison between the polypeptides of BPV-4 and other papillomaviruses showed that BPV-4 is evolutionarily closer to the epitheliotropic human and rabbit viruses than to BPV 1. PMID- 3039044 TI - Translational regulation in mouse hepatitis virus infection is not mediated by altered intracellular ion concentrations. AB - Infection of mouse L-2 fibroblasts with mouse hepatitis virus (MHV) results in strong inhibition of host cell protein synthesis. Since it has been suggested in other virus systems that translational control is modulated by changes in the intracellular ionic environment, we investigated the possible occurrence of similar changes during MHV infection. Membrane permeability to extracellular sodium ions was measured by culturing MHV-infected cells in the presence of 22Na+. Sodium influx into MHV-infected cells rose dramatically from 4 to 6 h post infection. This influx correlated chronologically with the expression of MHV mediated cell fusion. Cell fusion was blocked by the addition of a monoclonal antibody against the MHV E2 glycoprotein. This addition also resulted in a reduction in the normal influx of 22Na+, suggesting that E2 expression was responsible, directly or indirectly, for the increased permeability to sodium ions in infected cells. Cultures of MHV-infected cells were labelled with [35S]methionine in the presence of medium supplemented with sodium chloride at final concentrations ranging from 150 mM to 350 mM. Incorporation of radiolabel into proteins decreased with increasing NaCl concentration; however, the ratio of viral to cellular protein synthesis remained relatively constant. Similarly, alteration of intracellular Na+ and K+ levels by treatment of infected cells with ouabain had little effect on the pattern of viral/cellular protein synthesis. Using monoclonal anti-E2 antibody to inhibit Na+ influx, we demonstrated normal inhibition of host cell protein synthesis. We therefore conclude that MHV-induced shut-off of host translation is not mediated by changes in intracellular Na+ concentrations. PMID- 3039045 TI - Antigenic analysis of human and bovine parainfluenza virus type 3 strains with monoclonal antibodies. AB - The antigenic characteristics of eight human strains and two bovine strains, one of which is represented by two plaque variants, of parainfluenza virus type 3 were analysed. The strains and variants were compared using 52 monoclonal antibodies against five, two, six and six epitopes of the haemagglutinin neuraminidase (HN), fusion, nucleocapsid and matrix viral proteins respectively, employing radioimmuno-precipitation and immunofluorescence assays. The human strains, seven of which were isolated over 6 years at different geographical locations and the eighth one representing an older prototype strain, showed very little antigenic variation. Extensive differences were detected in all four proteins examined between the human strains and the two strains of bovine origin. Two bovine variants were less effectively neutralized than the prototype human strain with a series of monoclonal antibodies against the HN protein. PMID- 3039046 TI - Studies on the expression of spontaneous and induced interferons in mouse peritoneal macrophages by means of monoclonal antibodies to mouse interferons. AB - Monoclonal antibodies (MAbs) to mouse interferons (MuIFN) have been used to characterize the interferon-like activities spontaneously expressed in mouse peritoneal macrophages freshly explanted from normal pathogen-free mice. Injection of mice with MAbs to MuIFN-alpha or -beta resulted in a significant increase of vesicular stomatitis virus (VSV) multiplication in peritoneal macrophages. Addition of these MAbs to freshly explanted mouse macrophages accelerated the decay of the antiviral state to VSV during the 'ageing' in vitro of these macrophage cultures. Furthermore, these MAbs to MuIFN-alpha or -beta markedly inhibited the transfer of the antiviral state from freshly explanted peritoneal cells or macrophages to syngeneic macrophages 'aged' in vitro permissive for virus replication. These effects were not observed using a non neutralizing antibody to MuIFN-alpha, nor with a MAb to MuIFN-gamma. In all experiments sheep polyclonal antibodies to MuIFN-alpha/beta were more effective than the corresponding amount of MAbs to MuIFN-alpha or -beta. A mixture of both these MAbs was more effective than either alone. Interferons produced after stimulation of peritoneal macrophages with Newcastle disease virus (NDV) and of total peritoneal cells with lipopolysaccharides (LPS) have also been characterized by means of MAbs to IFNs. The results of neutralization studies with these antibodies indicated that MuIFN-beta was the major component of peritoneal cell IFN (induced by both NDV and LPS) and MuIFN-alpha was a minor component (13 to 17%). These data indicate that both MuIFN-alpha and -beta, but not MuIFN-gamma, are spontaneously present in/on mouse peritoneal macrophages and are produced after stimulation with NDV or LPS. PMID- 3039047 TI - Natural killer cells are not required for interferon-mediated prophylaxis against vaccinia or murine cytomegalovirus infections. AB - Natural killer (NK) cell-depleted or control mice were treated prophylactically with polyinosinic: polycytidylic acid (polyI: polyC) or purified beta interferon (IFN) and then infected with either vaccinia virus or murine cytomegalovirus. NK cell depletion alone enhanced virus titres in the spleen and peritoneal cavity. However, poly I: polyC and IFN inhibited virus replication equally well in control and NK cell-depleted mice. This suggests that prophylactic IFN treatment mediates antiviral effects independently of NK cells. PMID- 3039048 TI - Structure and variability of the a sequence in the genome of human cytomegalovirus (Towne strain). AB - We have defined the boundaries of the a sequence from human cytomegalovirus (CMV) strain Towne, characterized internal variability and determined the position of the cleavage site used to generate genomic termini. The cleavage site is positioned a fixed distance from two stretches of sequence homology that have been observed near the ends of many herpesvirus genomes. Unlike a comparable region in CMV (AD169), the CMV (Towne) a sequence has a relatively low level of variability within the a sequence and its structure is stable through repeated virus passage. PMID- 3039049 TI - Studies on DNA topoisomerases I and II in herpes simplex virus type 2-infected cells. AB - It has been suggested that herpes simplex virus (HSV) type 1 may induce a virus specific DNA topoisomerase activity which copurifies with virus-induced DNA polymerase. We have examined DNA topoisomerase (TOPO) I and II activities in HSV 2-infected HeLa S3 cells. Both activities were partially purified using DEAE cellulose, phosphocellulose and double-stranded DNA cellulose column chromatography. It was found that both activities could be separated from HSV-2 specific DNA polymerase. Throughout the purification TOPO I could be immunologically detected with a monoclonal antibody developed against human TOPO I. Regardless of the source, mock- or HSV-2-infected human cells, both types of topoisomerase were equally tolerant of 200 mM-KCl. There appeared to be no apparent heterogeneity of TOPO I in HeLa S3 cells through the course of the HSV-2 infection. We conclude that host cell topoisomerases are quite stable in HSV-2 infected HeLa S3 cells and that there is no evidence that HSV-2 is capable of inducing HSV-2-specific TOPO I and TOPO II activities. PMID- 3039050 TI - High genetic stability of the region coding for the structural proteins of yellow fever virus strain 17D. AB - The genome of the Pasteur 17D-204 vaccine strain of yellow fever virus has been cloned into pBR327. The inserts of recombinant plasmids were analysed by restriction cleavage pattern and compared with that of the genome of another substrain previously cloned and sequenced. Ten of the overlapping inserts were found to contain the sequence of the complete genome. We have sequenced approximately 3680 bases of the 5' region which codes for the C, M and E structural proteins and the NS1 non-structural protein. This sequence is the same as that reported previously, indicating a remarkable stability of these two vaccine substrains. PMID- 3039051 TI - Experimental Argentine hemorrhagic fever in rhesus macaques: virus-specific variations in pathology. AB - Two isolates of Junin virus (Espindola and Ledesma) inoculated into rhesus macaques produced distinct lesions which were strain-constant and similar to reported human cases of Argentine hemorrhagic fever. The Espindola isolate was associated with hemorrhagia, necrosis of bone marrow, and mild hepatocellular necrosis. Ledesma isolate was associated with pronounced polioencephalomyelitis and autonomic ganglioneuritis, but very mild or absent hepatocellular necrosis, bone marrow necrosis, and hemorrhagia. Deaths of Espindola-infected macaques were usually attributed to hemorrhagia with severe secondary bacterial infections, while in Ledesma-infected macaques, death was associated either with early severe secondary bacterial infections or slowly progressive polioencephalomyelitis. These two Junin virus isolates demonstrated hemorrhagic viscerotropism or neurotropism in macaques, suggesting that the variety of Argentine hemorrhagic fever syndromes in man may be virus-isolate determined. PMID- 3039052 TI - Treatment of cytomegalovirus pneumonitis with foscarnet (trisodium phosphonoformate) in patients with AIDS. AB - Foscarnet was administered to eight AIDS patients for suspected cytomegalovirus (CMV) pneumonitis as a continuous intravenous infusion for a minimum of 8 days. All the patients improved, three showing complete resolution of symptoms. Evidence of CMV infection from bronchoalveolar lavage samples was lacking in two patients. Adverse drug experiences consisted of thrombophlebitis, transient decreases in haemoglobin concentration, and reversible rises in serum creatinine levels. PMID- 3039053 TI - Effect of acyclovir on the uptake of 131I-labelled 1-(2'fluoro-2'-deoxy-beta-D arabinofuranosyl)-5-iodouracil in herpes infected cells. AB - Selective uptake of nucleoside analogues by herpes simplex virus infected cells may serve as the basis for a specific non-invasive diagnostic test for herpes simplex encephalitis. We have examined the effect of acyclovir on the selective uptake of [131I] 1-(2'-fluoro-2'-deoxy-beta-D-arabinofuranosyl)-5-iodouracil in herpes simplex virus infected primary rabbit kidney cells. Infected cells treated with acyclovir continued to concentrate [131I] 1-(2'-fluoro-2'-deoxy-beta-D arabinofuranosyl)-5-iodouracil for up to 24 h after the addition of the antiviral agent. These results indicated that therapy with acyclovir for as long as 24 h would not prevent the selective trapping of nucleoside analogues. This has important implications for the use of nucleoside analogues in diagnostic brain scans to detect herpes simplex encephalitis. PMID- 3039054 TI - Virus-specific factors in experimental Argentine hemorrhagic fever in rhesus macaques. AB - A nonhuman primate model for Argentine hemorrhagic fever has been developed that closely mimics the human clinical syndrome. Parenteral infection of adult Macaca mulatta with low-passage isolates of two Junin viral strains resulted in distinctive hemorrhagic or neurological disease in rhesus macaques that correlated with clinical illness patterns present in the humans from whom the viral strains were obtained. Transient leukopenia, together with thrombocytopenia and secondary bacterial septicemia, were documented among animals infected with both viral strains. In contrast, differing patterns of viremia, oropharyngeal viral shedding, and antibody response occurred in the two virus-infected groups. These results, together with postmortem virologic and histopathologic findings, suggest that viral-strain-specific factors are important determinants of clinical disease patterns in this model system. PMID- 3039055 TI - Affinity labeling of adenosine A1 binding sites. AB - The adenosine A1 receptors of sheep brain membranes have been identified by the specific binding of radiolabeled cyclohexyl[3H]adenosine ([3H]CHA). Pretreatment of membranes with periodate-oxidized CHA causes a dose- and time-dependent decrease in the number of binding sites. No decrease occurs when membranes are pretreated with CHA. Binding of [3H]CHA to the remaining sites occurs with the same characteristics as binding to the untreated receptor population. PMID- 3039056 TI - Interactions among lithium, calcium, diacylglycerides, and phorbol esters in the regulation of adrenocorticotropin hormone release from AtT-20 cells. AB - Interactions among lithium, calcium, and phorbol esters in the regulation of adrenocorticotropin hormone (ACTH) release were examined in a tumor cell line (AtT-20) of the anterior pituitary. Lithium, which blocks the phosphatase that converts inositol phosphates (IPs) to inositol, increases the levels of IPs in these cells and stimulates ACTH release. This ion potentiates the ability of calcium, an activator of phospholipase C, to raise levels of IPs in these cells and to stimulate ACTH secretion. Pretreatment of AtT-20 cells with calcium specifically abolishes the ACTH release response to lithium or calcium, a result suggesting that these secretagogues may act through a common mechanism to induce hormone secretion. Prior exposure of AtT-20 cells to either lithium or calcium also attenuates the ACTH release induced by phorbol ester, an activator of protein kinase C. To examine the link among lithium, calcium, phosphatidylinositol (PI) turnover, and phorbol ester-evoked ACTH secretion, AtT 20 cells were treated with 1-oleoyl-2-acetoyl-sn-3-glycerol (OAG), an analogue of the diacylgylcerols that are formed by phospholipase C during PI metabolism and that also activate protein kinase C. OAG itself does not alter ACTH release or the levels of IPs in AtT-20 cells. Pretreatment of AtT-20 cells with OAG, however, selectively blocks the ACTH release response to lithium, calcium, or phorbol ester. Furthermore, such pretreatment reduced the ability of lithium to increase levels of IPs. The results suggest that one mechanism of action of lithium is to potentiate selectively an action of calcium, possibly the stimulation of phospholipase C activity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3039057 TI - Autoradiographic localization of [3H]zolpidem binding sites in the rat CNS: comparison with the distribution of [3H]flunitrazepam binding sites. AB - The regional distribution of [3H]zolpidem, a novel imidazopyridine hypnotic possessing preferential affinity for the BZD1 (benzodiazepine subtype 1) receptor, has been studied autoradiographically in the rat CNS and compared with that of [3H]flunitrazepam. The binding of [3H]zolpidem to rat brain sections was saturable, specific, reversible, and of high affinity (KD = 6.4 nM). It occurred at a single population of sites whose pharmacological characteristics were similar to those of the benzodiazepine receptors labeled with [3H]flunitrazepam. However, ethyl-beta-carboline-3-carboxylate and CL 218,872 were more potent displacers of [3H]zolpidem than of [3H]flunitrazepam. The autoradiographic brain distribution of [3H]zolpidem binding sites was qualitatively similar to that previously reported for benzodiazepine receptors. The highest levels of [3H] zolpidem binding sites occurred in the olfactory bulb (glomerular layer), inferior colliculus, ventral pallidum, nucleus of the diagonal band of Broca, cerebral cortex (layer IV), medial septum, islands of Calleja, subthalamic nucleus, and substantia nigra pars reticulata, whereas the lowest densities were found in parts of the thalamus, pons, and medulla. Comparative quantitative autoradiographic analysis of the binding of [3H]zolpidem and [3H]flunitrazepam [a mixed BZD1/BZD2 (benzodiazepine subtype 2) receptor agonist] in the CNS revealed that the relative density of both 3H-labeled ligands differed in several brain areas. Similar levels of binding for both ligands were found in brain regions enriched in BZD1 receptors, e.g., substantia nigra pars reticulata, inferior colliculus, cerebellum, and cerebral cortex lamina IV. The levels of [3H]zolpidem binding were five times lower than those of [3H]flunitrazepam binding in those brain regions enriched in BZD2 receptors, e.g., nucleus accumbens, dentate gyrus, and striatum. Moreover, [3H]zolpidem binding was undetectable in the spinal cord (which contains predominantly BZD2 receptors). Finally, like CL 218,872 and ethyl beta-carboline-3-carboxylate, zolpidem was a more potent displacer of [3H]flunitrazepam binding in brain regions enriched in BZD1 receptors than in brain areas enriched in BZD2 receptors. The present data add further support to the view that zolpidem, although structurally unrelated to the benzodiazepines, binds to the benzodiazepine receptor and possesses selectivity for the BZD1 receptor subtype. PMID- 3039058 TI - Characterization of the dihydropyridine binding sites of rat neocortical synaptosomes and microvessels. AB - The dihydropyridine binding sites associated with rat neocortical synaptosomes and microvessels were compared using an in vitro [3H]PN 200-110 [(+)-[methyl-3H] isopropyl 4-(2,1,3-benzoxadiazol-4-yl)-1,4-dihydro-2,6-dimethyl-5- methoxycarbonylpyridine-3-carboxylate] binding assay. Saturation experiments yielded similar KD values (approximately 70 pM) and Bmax values (approximately 400 fmol/mg of protein) for the two membrane preparations. Interaction experiments with [3H]PN 200-110 and various calcium-modulating substances provided further evidence for the practically identical nature of the synaptosomal and microvascular dihydropyridine binding sites. These findings predict that lipophilic dihydropyridines, simultaneously occupying the two central binding sites, have the dual effect of altering neuronal function and local blood flow. PMID- 3039059 TI - Regulation of dopamine release from interplexiform cell processes in the outer plexiform layer of the carp retina. AB - The gamma-aminobutyric acid (GABA) antagonists bicuculline and picrotoxin stimulate a four- to fivefold increase in endogenous dopamine release from isolated intact carp retina. The release evoked by these agents is Ca2+ dependent, a finding suggesting a vesicular release. Using light microscopic autoradiography, we have localized the sites of dopamine release to the dopaminergic interplexiform cell processes of the outer plexiform layer, which synapse onto horizontal cells. Our findings support previous suggestions that the dopaminergic interplexiform cells receive GABAergic inhibitory input and that the effects of GABA antagonists on horizontal cells are mediated by dopamine release from the interplexiform cells. PMID- 3039060 TI - Chronic fatigue and myalgia syndrome: mitochondrial and glycolytic studies in skeletal muscle. AB - Clinical and biochemical findings in skeletal muscle in 11 patients with chronic fatigue myalgia syndromes of unknown aetiology are reported. All patients had severe asthenia for from one to 10 years with greatly limited exercise capacity and protracted exhaustion after minor exercise. Diffuse myalgia was prominent and was exacerbated for hours to days after exercise. Assay of skeletal muscle carnitine, phosphorylase, all glycolytic enzymes and the mitochondrial marker enzymes monoamine oxidase, isocitrate dehydrogenase and cytochrome oxidase were normal. These findings lend no support to the presence of a major defect in muscle intermediary energy pathways in this syndrome. PMID- 3039061 TI - Neurological involvement in X-linked hypophosphataemic rickets. AB - X-linked hypophosphataemic rickets is a familial form of Vitamin D resistant rickets in which gross bony and ligamentous changes may occur. Two patients showing severe spinal disease with evidence of spinal cord compression requiring neurosurgical intervention are reported. The management of such lesions may be problematic as cord compression may be found at several levels at presentation, and further difficulties develop after neurosurgical treatment. PMID- 3039063 TI - Vagus nerve and celiac ganglion lesions in generalized amyloidosis. A correlative study of familial amyloid polyneuropathy and AL-amyloidosis. AB - To clarify the cause of gastrointestinal disorders in systemic amyloidosis we made pathologic and morphometric studies of vagus nerves, celiac ganglia, stomach and rectum in three autopsied cases with type 1 familial amyloid polyneuropathy (FAP) and two with nonhereditary generalized amyloidosis (AL-amyloidosis). The gastric and rectal walls in all cases were affected in the same way by amyloid deposition. On the other hand, there was a great difference between the two diseases in the severity of vagus nerve and celiac ganglion lesions: the vagus nerves in FAP showed very extensive endoneurial deposition of amyloid with severe loss of myelinated nerve fibers, but in AL-amyloidosis there was no loss of myelinated nerve fibers and only slight amyloid deposition in the endoneurium. Similarly, in the celiac ganglion, intraganglionic deposition of amyloid was prominent in FAP and slight in AL-amyloidosis. It is known that bowel symptoms frequently occur in type I FAP and are less prominent in AL-amyloidosis. This study demonstrated that the gastrointestinal autonomic nerves were more markedly disturbed by amyloid in the former than in the latter, and disorder in neural control of the digestive tract may be responsible for the bowel symptoms in systemic amyloidosis, especially in type I FAP. PMID- 3039062 TI - Variant transthyretin in cerebrospinal fluid in familial amyloidotic polyneuropathy. AB - Structurally abnormal transthyretin is a precursor protein of amyloid fibrils in type I familial amyloidotic polyneuropathy (FAP). This variant transthyretin has an amino acid substitution of methionine for valine at position 30. The purpose of this study was to clarify whether this variant transthyretin also circulates in the cerebrospinal fluid (CSF) of patients with type I FAP. CSF transthyretin of the patients was purified and its primary structure determined. Sequence determination indicated that transthyretin consisted of a mixture of normal and variant transthyretin. Variant transthyretin was present in the CSF of all 5 Japanese FAP patients studied. The CSF concentration of variant transthyretin was high (0.72 +/- 0.15 mg/dl, mean +/- S.D.), suggesting that variant transthyretin is synthesized in the choroid plexus. Variant transthyretin was not present in any of 20 controls. The CSF concentration of total transthyretin in FAP patients was 1.74 +/- 0.42 mg/dl, which was not significantly different from controls. PMID- 3039064 TI - Platelet functions, alpha 2-adrenergic receptors and cytoplasmic free calcium are normal in the myotonic dystrophy of Steinert. AB - Platelet function tests were performed on 11 patients with myotonic dystrophy of Steinert (MD) and on 21 healthy control subjects. Using citrated platelet-rich plasma or washed platelets, MD patients had normal aggregation and secretion responses after stimulation with adrenaline, ADP, collagen, PAF, arachidonic acid and thrombin. Using intact and functional washed platelets, MD patients responded normally to adrenaline and had a similar affinity and number of alpha 2 adrenergic receptors as control patients as measured by [3H]dihydroergocryptine and [3H]yohimbine binding. In addition platelets from MD patients had normal basal and stimulated levels of cytoplasmic free Ca2+ as measured with the fluorescent Ca2+ probe quin2. Thus platelet functions, alpha 2-adrenergic receptors and cytoplasmic free Ca2+ are normal in MD. PMID- 3039065 TI - Inflammatory sensory polyradiculopathy and reactivated peripheral nervous system infection in a genital herpes model. AB - To study effects of herpesvirus reactivation on the nervous system, mice with latent genital herpes simplex virus type 2 (HSV-2) infections were immunosuppressed. Reactivated infection was detected by virus isolation or by antigen screening in histological sections of spines that contained cords, roots, and dorsal root ganglia. In ganglia, viral antigen was restricted to 2 distinct groups at T9-L2, and L6-S2 levels. In some ganglia, antigen was found in up to 4% of non-contiguous neurons, in their axons, and in endoneurial and satellite cells. Nerve roots distal to ganglia contained a few antigen-positive endoneurial cells and axons, but convincing antigen was not seen in proximal roots. Rare foci of anterior horn cells in the lower cord were the only central sites found to contain antigen. Comparison of antigen-containing ganglionic neuron counts to vaginal culture data indicate that peripheral virus shedding may depend on the number of neurons with reactivated infection. In non-immunosuppressed, latently infected mice, virus recovery was restricted to ganglionic explants from lower thoracic and lumbosacral regions; these ganglia contained no infectious virus in homogenates and no detectable antigen. These mice had an inflammatory polyradiculopathy in a similar distribution to virus found in latent and reactivated infection. The data show that genital HSV-2 infection can result in more extensive ganglionic latency and peripheral nervous system disease than has previously been recognized. Immunosuppression leads to reactivation and widespread, anatomically restricted antigen expression in many ganglia, consistent with the innervation of the genitourinary tract. Decalcified spine preparations provide a sensitive and simple way to detect virus reactivations and disease in these neural tissues. PMID- 3039066 TI - Quantification of rat sciatic nerve (Na+,K+)-ATPase by measurements of [3H]ouabain binding in intact nerve samples. AB - The (Na+,K+)-ATPase concentration in rat sciatic nerve was quantified by measurements of [3H]ouabain binding to intact nerve samples in a vanadate-Tris buffer. This gave values of 145-172 pmol/g wet weight in mature female rats. The methodological errors of the [3H]ouabain binding assay was identified, quantified and corrected for. The [3H]ouabain binding sites were homogeneous with respect to [3H]ouabain affinity with an apparent dissociation constant of 5 X 10(-8) mol/l, i.e. in the same range as in samples of skeletal muscles incubated at similar conditions. Post-mortem the [3H]ouabain binding site capacity in rat sciatic nerve decreased with a half-life time of 5 days, allowing measurements to be performed within 6 h after death with a reduction in [3H]ouabain binding sites of only 3%. When nerve samples were frozen the [3H]ouabain binding site concentration remained constant. An increase in sciatic nerve [3H]ouabain binding site concentration of 122% was seen from the 2nd to the 4th week of life followed by a decrease of 53% to the 12th week of life. In mature males the [3H]ouabain binding site concentration was 1.8 times that in nerves from age-matched female rats. Denervation caused a reduction of 61%, whereas K+ depletion or thyroid status has no effect on sciatic nerve [3H]ouabain binding site concentration. PMID- 3039067 TI - A rat model of herpes encephalitis with special reference to its potential for the development of diagnostic brain imaging. AB - A rat model of herpes encephalitis using intraocular inoculation of herpes simplex virus strain SC16 was investigated. Virus distribution in the brain was examined by virus isolation and immunocytochemical staining using immuno-gold silver and peroxidase-anti-peroxidase. At 5 days post-infection virus was found in the thalamus, hypothalamus, septum, colliculus, geniculate bodies, the pons, trapezoidium and medulla oblongata, but less frequently, in the cerebellum and occipital lobes. Possible routes of spread of virus and the potential of this model in neuro-radiological scanning procedures are discussed. PMID- 3039068 TI - Contrasting patterns of virus spread and neuropathology following microinjection of herpes simplex virus into the hippocampus or cerebellum of mice. AB - Two strains of herpes simplex virus type 1 (HSV) were microinjected into either the hippocampus or cerebellum of two different mouse strains. Immunoperoxidase staining revealed rapid and extensive spread of virus within the hippocampus and certain of its afferent connections. In contrast, only scattered peroxidase positive cells were observed following cerebellar inoculation. Extensive lesions were observed only in animals surviving intrahippocampal inoculations. These results suggest that localization of the pathological process in HSV encephalitis is due in part to a selective vulnerability of telencephalic or limbic structures. PMID- 3039069 TI - Teased fibre studies in leprous neuropathy. AB - Nerve biopsies were taken from four cases of leprosy, which included borderline tuberculoid, borderline and lepromatous types. They were examined in 1-micron resin sections and in teased preparations. The most common finding in teased fibres from each leprosy type was paranodal demyelination affecting successive internodes. In transverse sections some fibres showed changes associated with axonal atrophy. Together, these findings suggest that demyelination in some cases may be secondary to axonal changes. In addition, there was evidence of focal areas of demyelination affecting whole internodes of many fibres at the same level across the nerve and which was possibly caused by local factors. PMID- 3039070 TI - Controlled studies on the effects of alcohol ingestion on peripheral nerves of macaque monkeys. AB - In two different studies, monkeys were fed diets with 50% and 30% of calories replaced with alcohol for 5 and 3 years, respectively. Nerves were studied with electrophysiological and quantitative histological methods, but no deleterious effect of alcohol was identified. PMID- 3039072 TI - Time course and frequency dependence of synaptic vesicle depletion and recovery in electrically stimulated sympathetic ganglia. AB - The mammalian superior cervical sympathetic ganglion has been extensively used to study the kinetics of ACh metabolism and release. The present investigation examined the time course of changes in the number of synaptic vesicles and abundance of plasma membrane at preganglionic nerve terminals using stimulation protocols similar to those used in previous biochemical and electrophysiological studies. Continuous stimulation of the preganglionic trunk to the cat superior cervical ganglion in vivo produced an initially rapid fall in the number of clear synaptic vesicles followed by a subsequent plateau. Reciprocal changes in plasma membrane occurred with a similar time course. The plateau phase is interpreted as a steady-state where vesicle exocytosis is balanced by the rate of vesicle reformation from plasma membrane. During quiescent recovery, restoration of normal resting ultrastructure is initially rapid but slows with time as vesicle number and plasma membrane abundance approach pre-stimulation values, indicating that the rate of vesicle reformation at the end of stimulation is high and proportional to the number of vesicles incorporated into the plasma membrane. These results are interpreted as consistent with the 'vesicle hypothesis' of neurotransmitter release. PMID- 3039071 TI - The interpretation of electromyographic responses to electrical stimulation of the motor cortex in diseases of the upper motor neurone. AB - The complexities of interpreting results of electrical stimulation of the motor cortex in pathological states are discussed and illustrated by reference to results from a variety of patients with diseases affecting the upper motor neurone (multiple sclerosis, cervical spondylosis and myelopathy, motor neurone disease, hemiparesis due to cerebral infarction, and hereditary spastic paraplegia). The abnormalities of the electromyographic (EMG) responses after anodal cortical stimulation consisted of delay in the latency to onset, dispersion or reduction in response size or even absence of EMG responses. These changes were not confined to any specific condition or pathology. Previous work has suggested that the sequence of events that follow anodal cortical stimulation involves repetitive excitatory inputs to spinal motoneurones and transmission across at least one central synapse. Accordingly, delayed latencies may not exclusively indicate slowing of motor conduction, while the absence of any response may not indicate complete failure of conduction in corticomotoneurone pathways. PMID- 3039073 TI - Successful treatment of peripheral neuropathy with chemotherapy in osteosclerotic myeloma. AB - A patient with a severe polyneuropathy in association with osteosclerotic myeloma improved after melphalan and prednisone treatment. Aggressive treatment with chemotherapy is appropriate in this type of patient. PMID- 3039074 TI - Auditory brain-stem potentials studied with paired stimuli in normals. AB - Brain-stem auditory evoked potentials (BAEPs) to paired stimuli of various intervals were recorded using a subtraction technique to cancel the first and facilitate recognition of the second response. In a pilot series of 12 experiments, no latency change was encountered with paired click intervals (PCI) of 8 and 6 ms. Thirty healthy subjects were then investigated by presenting single and paired stimuli with a PCI of 4, 3.1, 2.3 and 1.5 ms at a repetition rate of 10/s to each subject. With diminishing PCIs, identification and latency measurement of waves IV and V became increasingly difficult because of marked amplitude reduction. Component III remained identifiable in all but two recordings and showed a statistically significant latency increase and amplitude reduction only when the stimuli were presented 1.5 ms apart. Component I did not show a consistent latency change with any PCI but its amplitude was significantly reduced with 4 ms PCI. These findings differ from those reported when using PCIs of 5 ms or more and when using single stimuli at a high repetition rate, where component V was the most stable. The alterations of evoked potentials found only with a PCI of 1.5 ms are probably of central origin. PMID- 3039075 TI - Total and split renal function in patients with renovascular hypertension: effects of angiotensin-converting enzyme inhibition. AB - Thirteen subjects with documented renovascular hypertension receiving chronic (greater than 1 month) therapy with an angiotensin-converting enzyme inhibitor (enalapril or captopril) underwent total and split renal function studies. Total glomerular filtration rate as assessed by inulin clearance was similar to that determined by radionuclide technique. Total effective renal plasma flow as assessed by p-aminohippurate clearance was lower than that determined by radionuclide technique. The glomerular filtration rate and effective renal plasma flow assessed by radionuclide technique of the stenotic kidney was comparatively lower than that of the non-stenotic kidney. No subject demonstrated complete loss of filtration or perfusion of the stenotic kidney. Five of six patients studied prospectively for 2 years have demonstrated stability of total renal function; the sixth patient, having a functional solitary stenotic kidney, has demonstrated stability of function following an initial abrupt decline in glomerular filtration rate and effective renal plasma flow. These results suggest that chronic angiotensin-converting enzyme inhibition therapy is not generally associated with near total absence of filtration of the stenotic kidney as has been suggested previously. Angiotensin-converting enzyme inhibitors may be safely and effectively utilized in the treatment of renovascular hypertension. PMID- 3039076 TI - Cardiac function as related to adrenergic activity in hypertensive left ventricular hypertrophy. AB - Studies in experimental animals and in hypertensive patients have shown that changes in cardiac anatomy and function are not just a simple consequence of the increased pressure load. The activity of the sympathetic nervous system is one of the factors that may influence cardiac performance and also, possibly, the cardiac anatomy of hypertensive patients. Animal studies have strongly suggested a possible role of adrenergic factors in the development of left ventricular hypertrophy. In man, plasma catecholamines are usually higher in hypertensive patients with left ventricular hypertrophy, and a correlation between left ventricular mass and plasma noradrenaline has also been observed. The decrease of cardiac performance after acute beta blockade has been found to be directly related to basal plasma noradrenaline concentration. An impaired response to beta adrenergic stimulation has been reported in hypertensive animals and has been confirmed in hypertensive patients with left ventricular hypertrophy. PMID- 3039077 TI - Electrical properties of phrenic motoneurons in the cat: correlation with inspiratory drive. AB - 1. Resting membrane potential (Vmp), input resistance (Rn), rheobase (Irh), and after hyperpolarization duration (AHPdur) and amplitude (AHPamp) were measured in 38 phrenic motoneurons of anesthetized, paralyzed, and artificially ventilated cats during hypocapnic apnea. The mean +/- SD and range of values for these variables were as follows: Vmp, -68 +/- 5mV (range: -60 to -82); Rn, 1.3 +/- 0.6 M omega (0.6-2.4); Irh, 9.7 +/- 5 nA (2-20); AHPdur, 68 +/- 19 ms (37-134); AHPamp, 3.3 +/- 1.8 mV (1.0-8.5). In 31 motoneurons, the membrane potential level at which firing occurred (Vthr) during intracellular current injection was measured. The mean value of Vthr was -58 +/- 3 mV (range: -52 to -64). 2. A histogram of Rn revealed a bimodal distribution. Also a plot of Irh against Rn showed a grouping of the motoneurons into two subpopulations: 1) low-Rn and high Irh cells, called type L neurons, and 2) high-Rn, low-Irh cells, called type H neurons. The overall negative linear correlation between Irh and Rn (r = -0.85; P less than 0.0001) resulted from this grouping rather than from a strictly linear relation between these two variables. 3. Electrical properties were compared for type L (n = 20) and type H (n = 18) phrenic motoneurons. The following mean values were found for each group, respectively: Rn, 0.8 and 1.8 M omega; Irh, 13.7 and 5.3 nA; AHPdur, 58 and 79 ms; AHPamp, 2.4 and 4.4 mV. All differences were significant (t test, P less than 0.001). Mean Vthr was the same for the two groups. 4. Comparison of these data with those available for lumbosacral motoneurons revealed that almost all investigated electrical properties of type L and type H phrenic motoneurons are similar to the analogous properties of type F (fast twitch) and type S (slow twitch) lumbosacral motoneurons, respectively. The apparent exception is the lower mean value of Irh for type L phrenic motoneurons compared with type F lumbosacral motoneurons. 5. For 13 cells, membrane potential was continuously monitored while spontaneous respiratory activity was restored by increasing CO2. It was found that at approximately the same end-tidal CO2 (about 7%) and a similar end-expiratory mean membrane potential level (approximately -70 mV), mean amplitude of peak inspiratory synaptic depolarization was higher in type H motoneurons (8.8 mV, n = 5) than in type L (2.9 mV, n = 8; P less than 0.001).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3039078 TI - Evidence for a fatigue-induced reflex inhibition of motoneuron firing rates. AB - 1. In previous studies on the adductor pollicis and biceps brachii muscles we suggested that motoneuron firing rates are inhibited by a reflex from the muscle during fatigue, since: the firing rates decline during a sustained maximal voluntary contraction (MVC); recovery of MVC firing rates is prevented if the fatigued state of the muscle is preserved for 3 min by local occlusion of its blood supply; and full recovery occurs during this time once the blood supply to the peripheral muscle is restored. These findings were confirmed in the present study for quadriceps contractions. 2. These results do not necessarily imply an inhibitory reflex. The lower firing rates recorded from the muscle fibers during an MVC following 3 min of postfatigue ischemia may have been caused by either reduced subject effort (decreased muscle activation by the CNS) or impaired peripheral impulse transmission under these conditions. The present experiments, carried out on the quadriceps and adductor pollicis muscles, were designed to test this alternative explanation. 3. For both muscles, MVC contractions were sustained for 40 s with a blood pressure cuff inflated to 200 mmHg. This was followed by 3 min ischemic rest and a second 20-s MVC before cuff release. Three minutes after the blood supply to the muscle was restored a third 20-s MVC was made. Single shocks were delivered to the muscle throughout to record twitches from the relaxed muscle (Tr) before and after each MVC, and any twitches super imposed on the voluntary contractions (Ts). The degree to which the muscle could be activated by voluntary effort was assessed from the ratio [1 - Ts/Tr]. For adductor pollicis, changes in the amplitude of the evoked M-waves were also measured. 4. Spike frequencies were only recorded during quadriceps experiments. These declined by 30% during the initial 40-s MVC. No recovery was seen in the second MVC following 3 min ischemic rest, but full recovery occurred within 3 min of cuff release. 5. Failure to retain full muscle activation was frequently seen in all three MVCs. However, for many well-motivated subjects twitch occlusion showed no reduction in the degree to which either the adductor pollicis or quadriceps muscles could be activated voluntarily during the MVC executed after 3 min of ischemic rest compared with that performed 3 min after the blood supply had been restored.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3039079 TI - Nonlinear directionally selective subunits in complex cells of cat striate cortex. AB - 1. We have analyzed receptive fields (RFs) of directionally selective (DS) complex cells in the striate cortex of the cat. We determined the extent to which the DS of a complex cell depends on spatially identifiable subunits within the RF by studying responses to an optimally oriented, three-luminance-valued, gratinglike stimulus that was spatiotemporally randomized. 2. We identified subunits by testing for nonlinear spatial RF interactions. To do this, we calculated Wiener-like kernels in a spatial superposition test that depended on two RF positions at a time. The spatial and temporal separation of light and dark bars at these two positions varied over a spatial range of 8 degrees and a temporal range of +/- 112 ms in increments of 0.5 degree and 16 ms, respectively. 3. DS responses in complex cells cannot be explained by their responses to single light or dark bars because any linear superposition of responses whose time course is uniform across space shows no directional preference. 4. Nonlinear interactions between a flashed reference bar that is fixed in position and a second bar that is flashed at surrounding positions help explain DS by showing multiplicative-type facilitation for bar pairs that mimic motion in the preferred direction and suppression for bar pairs that mimic motion in the null direction. Interactions in the preferred direction have an optimal space/time ratio (velocity), exhibited by elongated, obliquely oriented positive domains in a space-time coordinate frame. This relationship is inseparable in space-time. The slope of the long axis specifies the preferred speed, and its negative agrees with the most strongly suppressed speed in the opposite direction. 5. When the reference bar position is moved across the RF, the spatiotemporal interaction moves with it. This suggests the existence of a family of nearly uniform subunits distributed across the RF. We call the subunit interaction, as averaged across the RF, the "motion kernel" because its spatial and temporal variables are those necessary to specify the velocity, the only parameter that distinguishes a moving image from a temporally modulated stationary image. The nonlinear interaction shows a spatial periodicity, which suggests a mechanism of velocity selectivity for moving extended images.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3039080 TI - Autoradiographic differentiation of mu, delta, and kappa opioid receptors in the rat forebrain and midbrain. AB - While there is an abundance of pharmacological and biochemical evidence to suggest the existence of multiple opioid receptors, their precise localization within the brain is unclear. To help clarify this issue, the present study examined the distributions of the mu, delta, and kappa opioid receptor subtypes in the rat forebrain and midbrain using in vitro autoradiography. Mu and delta receptors were labeled with the selective ligands 3H-DAGO (Tyr- D-Ala-Gly-MePhe Gly-ol), and 3H-DPDPE (D-Pen2, D-Pen5-enkephalin), respectively, while the kappa receptors were labeled with 3H-(-)bremazocine in the presence of unlabeled DAGO and DPDPE. Based on previous findings in our laboratory, the labeling conditions were such that each ligand selectively occupied approximately 75% of each of the opioid sites. The results demonstrated that all 3 opioid receptor subtypes were differentially distributed in the rat brain. Mu binding was dense in anterior cingulate cortex, neocortex, amygdala, hippocampus, ventral dentate gyrus, presubiculum, nucleus accumbens, caudate putamen, thalamus, habenula, interpeduncular nucleus, pars compacta of the substantia nigra, superior and inferior colliculi, and raphe nuclei. In contrast, delta binding was restricted to only a few brain areas, including anterior cingulate cortex, neocortex, amygdala, olfactory tubercle, nucleus accumbens, and caudate putamen. Kappa binding, while not as widespread as observed with mu binding, was densely distributed in the amygdala, olfactory tubercle, nucleus accumbens, caudate putamen, medial preoptic area, hypothalamus, median eminence, periventricular thalamus, and interpeduncular nucleus. While all 3 opioid receptor subtypes could sometimes be localized within the same brain area, their precise distribution within the region often varied widely. For example, in the caudate putamen, mu binding had a patchy distribution, while delta and kappa sites were diffusely distributed, with delta sites being particularly dense ventrolaterally and kappa sites being concentrated ventromedially. These results support the existence of at least 3 distinct opioid receptors with possibly separate functional roles. PMID- 3039082 TI - Oxidative metabolism and glycolysis in benign brain tumors. AB - Glucose utilization in vivo and hexokinase activity and mitochondrial oxygen consumption in vitro were measured in a series of human brain tumors. Several relatively slow-growing tumors appeared to have depressed electron-transport activities coupled with a compensatory elevated glucose utilization. These data suggest that a decrease in oxidative metabolism and a corresponding increase in glycolysis are not necessarily correlated with malignancy in certain human brain tumors. PMID- 3039081 TI - Regulation of muscarinic acetylcholine receptor number in cultured neuronal cells by chronic membrane depolarization. AB - The effect of chronic membrane depolarization on the regulation of muscarinic acetylcholine receptor (mAChR) number was studied in neuroblastoma cells (clone N1E-115). Receptor number was determined by a filter binding assay using 3H quinuclidinyl benzilate (QNB) in membrane and crude cellular homogenates. Incubation with 50 microM veratridine (VTN), an activator of voltage-sensitive Na+ channels, induced a 50-200% increase in mAChR number at 24 hr, which was inhibited 80% by TTX. Scatchard analysis showed that affinity of the mAChR for 3H QNB was not affected by VTN. Upon withdrawal of VTN, mAChR number returned to control levels within 20 hr. Chronic membrane depolarization caused by incubation in medium containing 60 mM K+ induced a TTX-insensitive 50% increase in mAChR number at 24 hr. AChE activity was unaffected by chronic membrane depolarization. The VTN-induced increase in mAChR number was not blocked by coincubation with cycloheximide or tunicamycin, both inhibitors of de novo mAChR synthesis. The rate of mAChR degradation was reduced in the presence of 50 microM VTN, with the apparent half-life increased from approximately 18 hr (control) to approximately 40 hr (VTN). Although treatment with either 1 mM 8Br-cAMP or 1 mM 8Br-GMP failed to increase mAChR number, treatment with either the inorganic Ca2+ channel blocker Co2+ (1 mM) or the organic Ca2+ channel antagonist D600 (10-100 microM) produced 40-80% increases in mAChR number. The combination of VTN and either D600 or Co2+ failed to induce a greater increase in mAChR number than incubation with VTN alone.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3039083 TI - Serial PET studies of human cerebral malignancy with [1-11C]putrescine and [1 11C]2-deoxy-D-glucose. AB - Serial PET measurements of [1-11C]putrescine ([11C]PUT) uptake and glucose metabolic rate (GMR) using [1-11C]2-deoxy-D-glucose ([11C]2DG) were made on eight human subjects with a radiological and, in most cases, pathological diagnosis of primary or metastatic brain tumor. Blood-to-brain influx constants (Ki) were calculated for [11C]PUT. Tumor uptake of 11C after [11C]PUT injection was unidirectional peaking at 15 min. The mean +/- s.d. Kis for [11C]PUT for tumor and normal brain tissue were 0.78 +/- 0.045 and 0.024 +/- 0.007 ml cc-1 min-1, respectively (average of ratio, 3.11) whereas the ratio of GMR for tumor and normal brain tissue was 1.2 +/- 0.5. The mean Ki for four active, high grade astrocytomas was 0.098 +/- 0.030 in contrast to 0.027 +/- 0.008 ml cc-1 min-1 for two patients with low grade astrocytoma. Active high grade astrocytomas also showed marked CT contrast enhancement and regional glucose hypermetabolism. In one subject with brain metastases, both [11C]PUT and GMR correlated with a declining clinical picture in repeated studies over a 4-mo period. PET studies with [11C]PUT provide a better signal:noise ratio than GMR measurements, are useful for locating small glycolytically hypometabolic tumors and, when used in longitudinal studies in a single subject, appear to provide an index of degree of malignancy. PMID- 3039084 TI - Prevention of motion artifacts on dual isotope subtraction parathyroid scintigraphy. AB - A simple, effective technique is described to identify and eliminate motion artifacts which might potentially invalidate dual isotope subtraction parathyroid scintigraphy. Cobalt-57 markers, appropriately placed on the patient, allow detection of movement and permit realignment if movement occurs between imaging sequences. This technique should assure the accuracy of dual isotope parathyroid subtraction scintigraphy. PMID- 3039085 TI - Cerebral perfusion imaging. PMID- 3039086 TI - The influence of diets based on whole wheat, wheat flour and wheat bran on exocrine pancreatic secretion in pigs. AB - Six pigs, initially 40 kg live weight, were prepared with a duodenal pouch for collection of pancreatic juice to assess the influence of the type and amount of dietary fiber on exocrine pancreatic secretion. Four isonitrogenous diets were fed to each pig: W (whole wheat); WBS (whole wheat, wheat bran, wheat starch); WFS (whole wheat, wheat flour, wheat starch); and FC (wheat flour, wood cellulose). Wheat and wheat fractions came from the same batch of wheat. Crude fiber and dietary fiber (nonstarch polysaccharide) contents of diets W, WBS, WFS and FC (g/kg), respectively, were 40.8, 101.8; 63.7, 201.2; 20.5, 50.3; and 39.0, 60.7. Mean volumes of pancreatic juice (mL/24 h) for diets W, WBS, WFS and FC, respectively, were 4108, 4560, 2556 and 1757. Total enzymic and electrolyte concentrations were not significantly affected by diet changes, but mean protein and amylase outputs were lower for diet FC than for the others. It was concluded that the principal effect of increasing the dietary fiber content of the diets was to increase the volume of pancreatic juice and that this may have been due to noncellulosic components of the dietary fiber. PMID- 3039087 TI - Effect of dietary carbohydrates on glucagon and insulin receptors in genetically obese female Zucker rats. AB - Obese Zucker rats are hyperlipemic and mildly hyperglycemic. Because insulin and glucagon are involved in lipid and carbohydrate metabolism and they act via their receptors, we investigated the role of insulin and glucagon receptors in obese and lean female Zucker rats. Because dietary sucrose is more lipogenic than starch, we also studied the effect of dietary carbohydrates on the receptors. Significant phenotypic effect (obese greater than lean) was observed on plasma levels of glucose, triglyceride and insulin. Binding of insulin and glucagon to liver plasma membranes was significantly lower in obese rats than in lean rats. Lower insulin binding was due to a lower number of receptors as well as a lower affinity, whereas the lower glucagon binding was due only to a lower receptor number. Insulin binding in lean rats but not in obese rats was lower in sucrose fed than in starch-fed rats. Diet had no effect on glucagon binding. We propose that in obese Zucker rats, in addition to hyperinsulinemia, impaired glucagon activity as manifested by decreased glucagon binding to target tissues may be an important contributor to the hyperlipemia and obesity. PMID- 3039088 TI - Dietary fiber and cancer. PMID- 3039089 TI - Hydrolysis and synthesis of thiamin triphosphate in bacteria. AB - Thiamin triphosphate (ThTP) in early stationary phase cells of Escherichia coli grown in nutrient broth with 0.1% yeast extract was found to constitute approximately 5-7% of cellular thiamin diphosphate (ThDP) or around 5 nmol/g cell. Nearly the same level of ThTP was obtained in a Bacillus strain. When E. coli was loaded with an excess of ThTP or ThDP, cellular ThTP was found to be controlled in the course of the long term to maintain its ratio to the amount of cellular ThDP. The ThTP vs. ThDP ratio in E. coli cells after short-term ThDP uptake was found to be a function of the cellular growth phase. The ratio in early exponential phase E. coli cells was found to be approximately 4% and it became lower (less than 3%) when cell growth proceeded to the late exponential stage. Two phosphatases specific for ThTP (ThTPase) among thiamin phosphates were detected in E. coli. One required Mg2+ and was found mainly in the soluble fraction, while the other was Mg2+-independent and originated from the membrane. The two ThTPases were similar to their rat tissue counterparts. PMID- 3039090 TI - Up-regulation of alpha 2 adrenoceptors and down-regulation of beta 2 adrenoceptors by high-salt diet in normotensive men: enhanced up-regulation of operative (alpha 2:beta 2) adrenoceptor ratio predicts salt sensitivity. AB - In 24 normotensive male volunteers (age 20-25 years) reduction of sodium intake from 200 to 50 mmol/day over 2 weeks resulted in a 14% fall of alpha 2 adrenoceptors of platelets from 209.5 to 179.4 fmol/mg (P less than 0.01) and in a 16% rise of beta 2-adrenoceptors of lymphocytes from 13.3 to 16.2 fmol/mg (P less than 0.05) which was reversible by 2 weeks of high sodium intake. In contrast to the comparatively minor changes of alpha 2- and beta 2-adrenoceptor density, the functionally probably more relevant 'operative (alpha 2:beta 2) adrenoceptor ratio' decreased by 53% from 22.9 to 14.9 (P less than 0.01) during the low-salt diet. Although neither the individual changes of alpha 2- and of beta 2-adrenoceptor densities correlated with individual blood pressure changes induced by variations in sodium intake, there was a highly significant positive correlation (r = 0.55, n = 33; P less than 0.01) between the rises of the 'operative adrenoceptor ratio' and the rises of blood pressure induced by high salt intake. We conclude that the 'operative adrenoceptor ratio', although only determined on alpha 2- and beta 2- and not on alpha 1- and beta 1-adrenoceptors, and only on circulating blood cells, may be representative for sympathetic resistance vessel tone, at least as a function of variations of salt intake. Enhanced up-regulation of the 'operative adrenoceptor ratio' in the salt sensitive part of the population may be one important early step in the development of essential hypertension. PMID- 3039091 TI - Existence of renal alpha 1- and alpha 2-adrenoceptors in the human kidney: radioligand binding study in membranes from the human renal cortex and medulla. AB - We investigated alpha 1- and alpha 2-adrenoceptors by radioligand binding studies in plasma membranes from the human renal cortex and medulla using a new alpha 1 adrenoceptor antagonist, 3H-bunazosin, and an alpha 2-adrenoceptor antagonist, 3H rauwolscine. Human kidneys were obtained post-mortem (n = 4) or at surgery for carcinoma of the kidney (n = 10). The specific binding of both radioligands was rapid, saturable, reversible and specific. Scatchard analysis of 3H-bunazosin binding showed that the renal cortex had a KD of 4.6 nmol/l and a Bmax of 34.7 fmol/mg of protein, and the medulla a KD of 2.4 nmol/l and a Bmax of 23.2 fmol/mg. The specific binding of 3H-rauwolscine had a KD of 5.6 nmol/l and Bmax of 22.4 fmol/mg of protein for the cortex and values of 5.1 and 42.0, respectively, for the medulla. Competitive inhibition studies suggested that the binding of both radioligands was to sites with alpha 1 and alpha 2 specificity, respectively. The present studies demonstrate that human renal plasma membranes from both the cortex and medulla contain binding sites with both alpha 1- and alpha 2-adrenoceptors. PMID- 3039092 TI - Altered platelet phosphatidylinositol metabolism in essential hypertension. AB - The metabolism of phosphoinositides was investigated in platelet membranes from nine patients with essential hypertension (EHT) and from 10 age/sex-matched normotensive subjects. 32P-labelling or phosphatidic acid (PA), phosphatidylinositol (PI), phosphatidylinositol-4-phosphate (PIP) and phosphatidylinositol-4,5-bisphosphate (PIP2) was measured in lipid extracts prepared from resting 32P-prelabelled platelets. There were no differences in 32P PA between subject groups. In platelets of hypertensive patients, 32P incorporation into both PIP and PIP2 was greater (P at least less than 0.01). This increase apparently occurred at the expense of lower 32P-incorporation into PI (P less than 0.001). An alteration in the interconversion equilibrium between phosphoinositides which is directed towards polyphosphoinositide formation may lead to altered membrane ionic fluxes. PMID- 3039093 TI - The influence of the renin-angiotensin system on adrenergic vasoconstrictor responses in the forearm and splanchnic region in sodium deplete man. AB - In sodium deplete subjects forearm (FBF) and splanchnic (SBF) blood flows were measured at rest and during lower body negative pressure (LBNP) before and during angiotensin II (ANG II) blockade. Both FBF and SBF were reduced by LBNP. Forearm blood flow and forearm vascular resistance were unaffected by ANG II blockade (intra-arterial saralasin, n = 6; i.v. enalapril, n = 9) both at rest and during LBNP. In contrast, resting SBF increased and splanchnic vascular resistance decreased after i.v. enalapril (n = 10). The low resistance was completely unchanged during the following LBNP. It is concluded that acute ANG II blockade has no influence on the vascular resistance in the human forearm, but increases basal SBF in sodium depleted subjects and 'paralyses' the vasoconstrictor response to LBNP. PMID- 3039094 TI - Role of cyclic AMP in renin secretion by human transfected juxtaglomerular cells. AB - A human juxtaglomerular (JG) cell tumour was used for immortalization of renin secreting cells with three SV40 mutants. These transformed cells retained the same light and electron microscopic morphology as the human renal JG cells throughout subculture. Immunocytochemical staining showed the presence of renin in the elongated cells containing myofilaments and secretory granules. The renin produced was not stored within the cells but was released rapidly into the medium as prorenin. This permanent source of renin-producing cells was used for the study of renin regulation in vitro. Agents known to induce renin release such as dibutyryl cyclic AMP (cAMP), forskolin, isoproterenol and histamine were tested in cell culture. Forskolin induced renin secretion in a dose-dependent manner, as did dibutyryl cAMP. The JG cells responded to beta-agonists and to histamine by an H2 receptor. These results offer direct support for the hypothesis that cAMP is the second messenger in stimulation of renin secretion from human juxtaglomerular cells. PMID- 3039096 TI - Significance of donor transmitted disease in cardiac transplantation. AB - Infectious complications constitute a major cause of morbidity and mortality after heart transplantation. Those infections transmitted with the transplanted heart are potentially avoidable. Cytomegalovirus infections and toxoplasmosis occupy a prominent position in this category. The outcome of transplanting hearts from donors previously exposed to these infections depends largely on the existing immune status of the recipient. This paper is a retrospective study on the incidence of Toxoplasma gondii and cytomegalovirus infections in patients undergoing heart and heart-lung transplantation at Papworth Hospital, Cambridge. PMID- 3039095 TI - Hemodynamic effects of hypertonic sodium bicarbonate on denervated canine hearts. AB - The hemodynamic consequences of an intravenous bolus of hypertonic sodium bicarbonate were studied in normal dogs and in dogs that had undergone heart autotransplantation. Twenty-one autotransplanted dogs and seventeen sham-operated dogs were instrumented with an electromagnetic flow probe positioned around the ascending aorta. Hemodynamic data were collected three hours after the operation and then at daily intervals for one month, before and during sodium bicarbonate administration. The data indicates that hemodynamic base-line parameters remain lower in autotransplanted dogs than in the control animals during the whole length of the study. At the peak of the hypotensive response that follows the sodium bicarbonate injection the left ventricular performance is decreased to the same extent in both the autotransplanted and the control dogs. This negative inotropic response is enhanced in the presence of myocardial failure. In the autotransplanted group, reflex tachycardia was absent, resulting in a greater reduction in the cardiac index. These two effects explain the greater reduction of arterial pressure in the autotransplanted dogs. PMID- 3039097 TI - Quantitative analysis of immunosuppression in cyclosporine-treated heart transplant patients with lymphoma. AB - Lymphoma remains an important complication after heart transplantation in the era of cyclosporine immunosuppression. It has been generally assumed that the occurrence of lymphoma related to quantitative degree of immunosuppression, and this assumption is exemplified in the treatment of some such lymphomas with reduction of immunosuppression. Seventy-five consecutive survivors of heart and heart-lung transplantation between December 1980 and July 1983 were treated with cyclosporine and steroids; some received rabbit anti-thymocyte globulin for prophylaxis or treatment of rejection. Measured quantitative parameters of immunosuppression during the first three months after transplantation included mean cyclosporine level, total rabbit anti-thymocyte globulin dosage, number of days of T-cell suppression, and mean cyclosporine level during T-cell suppression. Serial Epstein-Barr virus antibody titers were measured. Lymphoma was diagnosed in six patients. The mean number of episodes of rejection did not differ between the lymphoma and the non-lymphoma groups. All quantitative measures of immunosuppression were higher in the lymphoma group, but this difference achieved statistical significance only in the case of total dosage of rabbit anti-thymocyte globulin (p less than 0.02). Four of the six lymphoma patients received some or all of their rabbit anti-thymocyte globulin dosage as prophylaxis against rejection; one received rabbit anti-thymocyte globulin solely for rejection; and the sixth received no rabbit anti-thymocyte globulin. Ebstein Barr virus titer conversion (four-fold rise in titer) alone was not significantly associated with occurrence of lymphoma. However, logistic regression analysis suggested that Epstein-Barr virus conversion in concurrence with high mean cyclosporine levels predicted a higher risk of lymphoma.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3039098 TI - 'Rejection or infection' predictive value of T-cell subject ratio, before and after heart transplantation. AB - Peripheral T-cell subsets were monitored in ten heart and two heart-lung recipients pre- and up to one year post-operatively. Prior to transplantation four patients had T-helper/T-cytotoxic suppressor ratios (TH/TS-C) above the range for normal healthy controls and all required treatment for rejection episodes, as compared with three of eight patients whose pre-transplantation ratios were within the normal range. No patient with high TH/TS-C ratios developed cytomegalovirus infection as compared with all of the eight patients with normal ratios. Post-transplantation cytomegalovirus infection was the major cause of alterations in TH/TS-C ratios. T-cell subset inversion always preceded the diagnostic rise in cytomegalovirus antibody titre in both primary and secondary cytomegalovirus infections. Inversion was also noted with Pneumocystis carinii infection. Reversal of the TH/TS-C ratio was due to a major increase in the absolute numbers of TS-C cells and was usually followed by a rise in the number of cells expressing a natural killer cell phenotypic marker (Leu-7). All patients with primary cytomegalovirus and two of four cases with secondary cytomegalovirus retained inversion throughout follow-up and showed significantly increased numbers of TS-C and Leu-7 bearing cells. However, the absolute numbers of TH fell by 200 days after transplantation in all patients irrespective of their TH/TS-C ratio. Although TH/TS-C ratio inversion was a predictor of cytomegalovirus infection, no association was found between changes in T-cell subsets after transplantation and rejection episodes. PMID- 3039099 TI - Uptake of myocardial imaging agents by rejected hearts. AB - Technetium 99 m pyrophosphate, Gallium 67 and Thallium 201 uptakes were measured in heterotopically transplanted rat hearts. Five days after transplantation, Technetium 99 m pyrophosphate, and Gallium 67 uptakes were significantly higher in allogeneic grafts than in syngeneic grafts. At an early stage of rejection (three days after transplantation), only Technetium 99 m pyrophosphate uptake in the left ventricle of allogeneic grafts showed a significant difference (p less than 0.04). At five days, Thallium 201 uptake was significantly lower in allo- than syngeneic grafts. There was a positive correlation between radionuclide uptake and histologic degree of rejection for Technetium 99 m pyrophosphate and Gallium 67 while Thallium 201 uptake correlated negatively. Analysis of variance revealed that hearts with no or minimal rejection had statistically different uptakes than hearts with mild to moderate rejection. These results suggest that uptake of imaging agents might be useful in the diagnosis of rejection of the transplanted heart. PMID- 3039100 TI - Nuclear magnetic resonance and proton relaxation times in experimental heterotopic heart transplantation. AB - It should be possible to detect heart transplant rejection by nuclear magnetic resonance (NMR) imaging if it induces myocardial T1 and T2 proton relaxation time alterations or both. We studied 20 Lewis rats after a heterotopic heart transplantation. In vitro measurement of T1 and T2 was performed on a Minispec PC20 (Bruker) 3 to 9 days after transplantation. Histologic analysis allowed the quantification of rejection process based on cellular infiltration and myocardiolysis. Water content, a major determinant of relaxation time, was also studied. T1 and T2 were significantly prolonged in heterotopic vs orthotopic hearts (638 +/- 41 msec vs 606 +/- 22 msec for T1, p less than 0.01 and 58.2 +/- 8.4 msec vs 47.4 +/- 1.9 msec for T2, p less than 0.001). Water content was also increased in heterotopic hearts (76.4 +/- 2.3 vs 73.8 +/- 1.0, p less than 0.01). Most importantly, we found close correlations between T1 and especially T2 vs water content, cellular infiltration, and myocardiolysis. We conclude that rejection reaction should be noninvasively detected by NMR imaging, particularly with pulse sequences emphasizing T2. PMID- 3039101 TI - Bicarbonate-dependent solute transport in kidney tubules. PMID- 3039102 TI - Lymphocyte abnormalities in infants born to drug-abusing mothers. AB - To evaluate the possible effects of maternal intravenous drug use on infant immunity, we measured the in vitro peripheral blood mononuclear cell proliferative responses to phytohemagglutinin (PHA) and pokeweed mitogen, T cell subset numbers, immunoglobulin levels, and titers of antibodies to cytomegalovirus (CMV) and human immunodeficiency virus (HIV) in a group of drug abusing mothers and their infants. Infants of drug abusers had a lower proliferative response to mitogen, associated with altered kinetics of the maximum response to PHA. The OKT4/OKT8 ratio decreased with age in the drug exposed infants compared with control infants (P less than 0.005). There was no evidence of CMV infection in either group. One mother and her infant had antibody to HIV. Our data demonstrate that infants of intravenous drug-using mothers have distinct immunologic differences at birth compared with non-drug-exposed infants and that these persist throughout the first year of life. The cause appears unrelated to intrauterine viral infection, suggesting a direct toxic effect of the drugs on fetal immunologic development. PMID- 3039103 TI - Klippel-Trenaunay syndrome and other cases of lower limb hypertrophy: pediatric surgical implications. AB - We report 16 cases of lower limb hypertrophy seen in the past 5 years; of these, four had a typical picture of Klippel-Trenaunay syndrome. We discuss diagnostic modalities, complications, treatment, and outcome. Special emphasis is placed on the treatment with intermittent pneumatic compression and the importance of an organized approach to the condition. PMID- 3039104 TI - Psychobiological factors predicting the course of breast cancer. AB - Drawing on a carefully controlled sample of 52 women with a history of breast carcinoma and 34 healthy controls, this prospective study examined empirical associations between psychological factors and the progression of neoplastic disorders over a follow-up period averaging 624 days. Psychological variables were psychometrically assessed by self-report measures. A multiple regression analysis which controlled for disease stage at original diagnosis, age, total length of disease course, hematological factors, and blood chemistries measured at study onset showed neoplastic spread to be associated with a repressive personality style, reduced expression of negative affect, helplessness hopelessness, chronic stress, and comforting daydreaming. The identified model of medical and psychological variables accounted for 56% of the observed variance. A psychobiological model of brain-body disregulation provided the best account of the observed associations between psychological functioning and the progression of disease. Future research is necessary to examine the role which psychological functioning may exert upon health-relevant behaviors that might blunt the benefits of professional health care. PMID- 3039105 TI - Synovial sarcoma. PMID- 3039106 TI - Asymmetric catecholimidazolines and catecholamidines: affinity and efficacy relationships at the alpha adrenoreceptor in rat aorta. AB - The epinephrine (EPI) stereoisomers interact with the alpha adrenoreceptor according to the Easson-Stedman model with an order of potency of (-)-EPI greater than (+)-EPI = Epinine. A series of catecholimidazolines (CI) and catecholamidines (CA) were compared with the EPI series for the relationship of stereoisomerism to potency, affinity and efficacy. Within each group of desoxy compound and stereoisomers obtained by -OH substitution at the benzylic position, differences in potency were found to be due solely to differences in affinity; differences in efficacy were not significant. The stereoisomers of the CI and CA series followed the order of potency predicted by the Easson-Stedman model: (-) isomer greater than (+)-isomer. The desoxy analogs, in contrast to the prediction based on the Easson-Stedman hypothesis, were equal (CI) or greater (CA) in potency than the more potent (-)-isomer of each series. Possible explanations for this include differences in physical properties in the desoxy analogs of CI and CA compared with the corresponding enantiomers. Methyl or benzyl substitution at C-4 of the imidazoline ring decreased potency over 100-fold; potency differences between enantiomers were negligible. Thus, the Easson-Stedman model cannot be extended to either the CI or CA series of alpha adrenoreceptor agonists. PMID- 3039107 TI - A nonglucocorticoid steroid analog of methylprednisolone duplicates its high-dose pharmacology in models of central nervous system trauma and neuronal membrane damage. AB - Prior studies have demonstrated that intensive treatment with high doses of methylprednisolone (MP) can beneficially affect the acutely injured central nervous system by a variety of mechanisms and promote neurological recovery in experimentally injured animals. In view of the fact that these actions are associated only with MP doses greatly in excess of those required for classical glucocorticoid receptor-mediated actions of the steroid, the possibility was examined that this high-dose pharmacology of MP could be duplicated by a nonglucocorticoid analog. Accordingly, U-72099E (17,21-dihydroxy-11 alpha-t butylacetoxy-1,4-pregnadiene-3,20-dione- 21-hemisuccinate, sodium salt) was synthesized and tested for its ability to duplicate the high-dose effects of MP in a concussive head injury model in mice and in an in vitro model of lipid peroxidation-induced membrane damage using rat brain synaptosomes. The absence of glucocorticoid-related activity of U-72099E was confirmed by its inability to either suppress body weight gain or cause thymic involution in mice treated with doses up to 100 mg/kg/day for 4 days. On the other hand, MP at 30 mg/kg/day for 4 days caused a complete inhibition of body weight gain and a 43.5% reduction in thymus weight. Moreover, U-72099E, at concentrations of 10(-5) M or lower, failed to suppress adrenocorticotropin secretion by mouse AtT-20 pituitary cells in culture, whereas dexamethasone or MP at concentrations of 10(-6) M and lower caused a marked suppression in adrenocorticotropin secretion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3039108 TI - Naloxonazine effects on the interaction of enkephalin analogs with mu-1, mu and delta opioid binding sites in rat brain membranes. AB - The authors have characterized the opioid receptors of rat brain membranes using self- and cross-displacement studies with both tritiated and unlabeled [D-Ala2, D Leu5]-enkephalin and [D-Ala2, MePhe4, Gly-ol5]-enkephalin. Mathematical modeling demonstrated the presence of three classes of binding sites, corresponding to mu, delta and the putative mu-1 classes of site. Unlabeled naloxonazine shows high affinity for all three classes of sites, with highest affinity for the mu-1 sites. Membranes were preincubated with 50 nM naloxonazine or with controls (50 nM naloxone or buffer) for 30 min. Preincubation of membranes with 50 nM naloxonazine resulted in a dramatic, nearly 2-fold reduction in the binding of [3H][D-Ala2, D-Leu5]-enkephalin and [3H][D-Ala2, MePhe4, Gly-ol5]-enkephalin relative to the controls. Quantitative analyses using mathematical modeling with program "LIGAND" suggested that this effect was primarily "competitive," i.e., attributable to changes in affinity, with no apparent or detectable noncompetitive or irreversible effects on binding capacities for the three classes of sites. PMID- 3039109 TI - Enhanced pulmonary alpha-2 adrenoceptor responsiveness under conditions of elevated pulmonary vascular tone. AB - Alpha-1 and alpha-2 adrenoceptor-mediated vasoconstriction was studied in the in situ, autoperfused pulmonary circulation of the open-chest anesthetized dog under conditions of normal and elevated pulmonary vascular tone. Under conditions of normal pulmonary vascular tone (10 +/- 1 mm Hg), methoxamine, a selective alpha-1 adrenoceptor agonist, and B-HT 933, a selective alpha-2 adrenoceptor agonist, elicited maximal increases in lobar perfusion pressure of 5 and 2 mm Hg above resting pulmonary tone, respectively. When pulmonary vascular tone was elevated progressively with the thromboxane mimetic, U-46619, serotonin or PGF2 alpha, alpha-1 adrenoceptor-mediated pulmonary vasoconstrictor responses to methoxamine were unaffected, whereas alpha-2 adrenoceptor-mediated pulmonary pressor responses to B-HT 933 were enhanced. Overall the response to B-HT 933 was enhanced 4-fold when pulmonary perfusion pressure was elevated to 19.8 +/- 0.8 mm Hg with U-46619 and almost 5-fold when elevated to 27.0 +/- 1.2 mm Hg. Pulmonary vasoconstrictor responses to angiotensin II were unaffected by elevated pulmonary vascular tone. Enhanced responsiveness of B-HT 933 to elevated pulmonary vascular tone was antagonized by the selective alpha-2 adrenoceptor antagonist, rauwolscine (100 micrograms/kg i.v.), and unaffected by the selective alpha-1 adrenoceptor antagonist, prazosin (100 micrograms/kg i.v.). When canine intralobar pulmonary veins were studied in vitro they contracted to B-HT 933 whereas intralobar pulmonary arteries did not respond. These data indicate that alpha-2 adrenoceptor responsiveness is enhanced markedly and selectively under conditions in which pulmonary vascular tone is elevated. PMID- 3039110 TI - Muscarinic acetylcholine receptor-mediated phosphoinositide turnover in cultured cerebellar granule cells: desensitization by receptor agonists. AB - Cultured granule cells prepared from 8-day postnatal rats were used for the study of carbachol-induced phosphoinositide turnover. The addition of carbachol induced a 20- to 30-fold increase in [3H]inositol monophosphate (IP1) accumulation within 30 min in the presence of 20 mM LiCl in granule cells prelabeled with [3H]myoinositol. Carbachol also stimulated the formation of [3H]inositol bisphosphate and [3H]inositol trisphosphate assayed either in the presence or in the absence of lithium; the increase in [3H]inositol triphosphate and [3H]inositol biphosphate synthesis appeared to be faster in the time course but much smaller in amount when compared with the formation of [3H]IP1. The EC50 of carbachol was approximately 7 microM, and the saturation concentration was about 100 microM. This carbachol-induced response was completely blocked by two muscarinic acetylcholine receptor (mAChR) antagonists, atropine and pirenzepine, with a Ki of 0.5 and 120 nM, respectively. Saturation studies of the binding of [3H]N-methylscopolamine and [3H]quinuclidinyl benzilate to mAChRs in intact granule cells revealed the presence of a single class of binding sites with a Kd of 140 and 126 pM, respectively, and a Bmax of approximately 70 fmol/10(6) cells for both ligands. Within 1 hr after pre-exposure of cells to carbachol, the subsequent carbachol-induced [3H]IP1 accumulation was reduced by about 50%, whereas mAChR binding, assessed by using either [3H]N-methylscopolamine or [3H]quinuclidinyl benzilate, was unchanged. A second slower phase of desensitization was associated with a loss of mAChR binding sites; at 18 hr, the decrease of responsiveness was about 80%, whereas the loss of mAChR sites was about 60%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3039111 TI - Differential antagonism of alpha-1 and alpha-2 adrenoceptor-mediated pressor responses by urotensin I, a selective mesenteric vasodilator peptide. AB - In order to characterize the hemodynamic actions of urotensin I, a vasodilator peptide with selectivity for the mesenteric vascular bed, we studied its hypotensive effects and interference with alpha-1 and alpha-2 adrenergic vasoconstrictor responses in the rat. After i.v. administration in anesthetized rats, urotensin I (0.06-6 nmol/kg) produced a dose-dependent lowering of arterial blood pressure. At hypotensive doses, urotensin I was about 3 times more potent in antagonizing systemic pressor responses to the selective alpha-1 adrenoceptor agonist, phenylephrine, than responses to the nonselective adrenoceptor agonist, norepinephrine. Additional studies were performed on the blood-perfused mesenteric bed of the anesthetized rat and on the isolated rat superior mesenteric artery, using as tools phenylephrine, norepinephrine and the relatively selective alpha-2 adrenoceptor agonist, alpha-methylnorepinephrine. The selectivity of the three agonists for vascular alpha-1 and alpha-2 adrenoceptors in the blood-perfused mesenteric bed was confirmed using prazosin and yohimbine as selective antagonists of alpha-1 and alpha-2 adrenoceptors, respectively. Urotensin I diminished the maximum increase in perfusion pressure and shifted the log dose-response curves to the right for all three agonists. A marked selectivity of urotensin I for alpha-1 adrenoceptor-mediated responses was observed: IC30 values of the peptide for pressor responses to phenylephrine, norepinephrine and alpha-methylnorepinephrine were 0.05, 0.83 and greater than 6 nmol/kg, respectively. A less pronounced selectivity of urotensin I for alpha-1 adrenoceptor-mediated contractions could be demonstrated in isolated strips of the superior mesenteric artery of the rat.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3039113 TI - On the relationship between clonidine hypotension and brain beta-endorphin in the spontaneously hypertensive rat: studies with alpha adrenergic and opiate blockers. AB - The relationship between the centrally mediated hypotensive and bradycardic effects of clonidine to central alpha-2 adrenergic receptor activation, brain beta-endorphin (BE) release and opiate receptor activation was studied in chloralose-anesthetized spontaneously hypertensive rats (SHRs) and Wistar-Kyoto rats, using a cerebroventricular perfusion system. Prior treatment of SHRs with i.v. naloxone (2 or 4 mg/kg) or i.c.v. yohimbine (10 or 20 micrograms/kg) reduced the hypotension and bradycardia induced by i.c.v. clonidine, but in Wistar-Kyoto rats naloxone had no similar blocking effects. Prazosin (20 micrograms/kg i.c.v.) reduced the clonidine bradycardia but not the hypotension in SHRs. Hypotension in the SHRs due to i.c.v. alpha-methylnorepinephrine (20 micrograms/kg) was reduced by both naloxone and yohimbine whereas alpha-methylnorepinephrine bradycardia was reduced by yohimbine but not by naloxone. Prior hypothalamic lesions in the SHRs reduced clonidine hypotension, but not bradycardia, and interfered with naloxone blockade of the residual clonidine hypotensive effect. Clonidine lowered immunoreactive BE levels in SHR hypothalamus, medulla and pituitary but did not change BE levels in the i.c.v. perfusate. The findings support the idea that in the SHRs, clonidine hypotension results from alpha-2 adrenergic stimulation of brain, causing BE release and central opiate receptor activation, and they suggest that the hypothalamus is involved in these interactions. Also, clonidine hypotension and bradycardia appear to involve different mechanisms in brain. PMID- 3039112 TI - Kappa agonist-induced diuresis: evidence for stereoselectivity, strain differences, independence of hydration variables and a result of decreased plasma vasopressin levels. AB - Marked diuresis has previously been reported after administration of kappa opioid agonists. The present study shows that this effect is stereospecific; MR-2034 markedly increased urinary output over the dose range 0.08 to 1.25 mg/kg, whereas the opposite isomer, MR-2035, was markedly less potent. Bremazocine increased urinary output in Long-Evans hooded and Sprague-Dawley albino rats as well as lean and fatty Zucker rats. In the lean Zucker and the albino rats, bremazocine produced an inverted U-shaped diuretic dose-effect curve, an effect characteristic of kappa agonists with mu agonist activity. This pattern was not seen with the fatty Zucker rats or the Long-Evans hooded rats. The full kappa agonists bremazocine, ethylketazocine and U-50,488 increased urinary output under three different conditions of hydration: water loaded, normal hydration and water deprived. In contrast, the partial kappa agonists reliably only increase urinary output under the normal hydration condition. The diuretic effects of full and partial kappa agonists correlated with plasma vasopressin levels in water deprived rats. The full kappa agonists (ethylketazocine, U-50,488, tifluadom and MR-2034) suppressed plasma vasopressin levels below the threshold of detectability of the radioimmunoassay, whereas the partial kappa agonists (nalorphine and butorphanol) suppressed vasopressin levels compared with control values but did not have the efficacy of the full kappa agonists. All these results support the hypothesis that kappa agonists produce their diuretic effect by suppression of plasma vasopressin levels. PMID- 3039114 TI - CGS 9896 and ZK 91296, but not CGS 8216 and RO 15-1788, are pure benzodiazepine receptor antagonists on mouse neurons in culture. AB - The effects of several benzodiazepine receptor ligands on gamma-aminobutyric acid (GABA) responses and on diazepam (DZ) enhancement of GABA responses on mouse spinal cord and cerebral hemisphere neurons are reported. The neurons were grown in primary dissociated cell culture, and intracellular microelectrode recording techniques were used. DZ and Ro 15-1788 reversibly enhanced GABA responses from spinal cord neurons in a concentration-dependent manner. Maximal enhancement was obtained for DZ at 500 nM (82%) and for Ro 15-1788 at 1 microM (18%). CGS 9896 and ZK 91296, in concentrations from 1 nM to 10 microM, were devoid of intrinsic effects in spinal cord and cerebral hemisphere neurons. CGS 8216 and methyl 6,7 dimethoxy-4-ethyl-beta-carboline-3-carboxylate (DMCM) reversibly and dose dependently reduced GABA responses on spinal cord neurons. Maximal reduction was obtained for CGS 8216 at 50 nM (10%) and for DMCM at 1 microM (39%). CGS 8216 (IC50 = 2.6 nM), ZK 91296 (IC50 = 9 nM), CGS 9896 (IC50 = 17 nM) and Ro 15-1788 (IC50 = 40 nM) antagonized (100 nM) DZ enhancement of GABA responses in a concentration-dependent manner. In addition, CGS 9896 antagonized the effects of Ro 15-1788 and CGS 8216 on GABA responses, and CGS 9896 and ZK 91296 antagonized the reduction of GABA responses by DMCM. On the studied neurons, DZ can be classified as an agonist, Ro 15-1788 as a weak partial agonist, CGS 8216 as a weak partial inverse agonist and DMCM as an inverse agonist at the benzodiazepine receptor. CGS 9896 and ZK 91296 were pure antagonists at the benzodiazepine receptor. PMID- 3039115 TI - Preclinical evaluation of McN-5707 as a potential antidepressant. AB - Based on its activity in a variety of tests in vivo and in vitro McN-5707 [trans 6-(2-chlorophenyl)-1,2,3,5,6,10b-hexahydropyrrolo- (2,1-a)isoquinoline] is a novel potential antidepressant. McN-5707 blocked tetrabenazine-induced sedation and ptosis in mice and rats, and potently inhibited the uptake of norepinephrine by synaptosomes from rat hypothalamus (Ki approximately 2 nM), and the uptake of serotonin by synaptosomes from rat cerebral cortex (Ki approximately 10 nM). McN 5707 also inhibited the uptake of dopamine by synaptosomes from rat striatum (Ki approximately 40 nM); however, the stereotypic behavior often caused by dopamine uptake inhibitors was not evident in rats at doses of 300 mg/kg (p.o.) or less. In receptor binding assays, McN-5707 potently inhibited ketanserin binding to serotonin 5-HT2 receptors in synaptic membranes from rat cerebral cortex (apparent Ki approximately 8 nM). In mice, McN-5707 antagonized 5 hydroxytryptophan-induced head twitches. Spiperone binding to dopamine D2 receptors in synaptic membranes from rat striatum was weakly inhibited by McN 5707 (apparent Ki approximately 400 nM), as was the binding of WB4101 to alpha-1 adrenergic receptors (apparent Ki approximately 150 nM). McN-5707 was essentially inactive as an inhibitor of [3H]clonidine binding to alpha-2 adrenergic receptors and of [3H]quinuclidinyl benzilate binding to muscarinic receptors. In experiments with guinea pig ileum, McN-5707 weakly antagonized histamine-induced contractions and exhibited virtually no cholinergic or anticholinergic activity. Our observations indicate McN-5707 possesses attributes of both tricyclic and newer atypical antidepressants because it inhibits the uptake of both norepinephrine and serotonin, and blocks 5-HT2 receptors, but lacks some of the anticholinergic and behavioral properties often associated with them. PMID- 3039117 TI - Evaluation of the alpha and beta adrenoceptor-mediated activities of the novel, orally active inotropic agent, ibopamine, in the cardiovascular system of the pithed rat: comparison with epinine and dopamine. AB - The alpha and beta adrenoceptor-mediated effects of the novel, orally active inotropic prodrug, ibopamine, have been studied in the pithed rat and compared with those effects mediated by dopamine and the active form of ibopamine, epinine. All three agents produced alpha adrenoceptor-mediated pressor responses in pithed rats, and the vasopressor effects of ibopamine and epinine, but not dopamine, were potentiated by beta adrenoceptor blockade with propranolol (3 mg/kg i.v.). Catecholamine depletion with reserpine (5 mg/kg i.p.) did not affect the vasoconstrictor response elicited by any of these agents, indicating a direct effect in the vasculature. Epinine was 10 times more potent than ibopamine or dopamine. The pressor response to all three agents was antagonized by the alpha-1 adrenoceptor antagonist, prazosin (0.1 mg/kg i.v.) and the alpha-2 adrenoceptor antagonist, rauwolscine (0.5 mg/kg i.v.), suggesting the involvement of both alpha adrenoceptor subtypes in the vasopressor responses elicited by these compounds. After complete blockade of alpha adrenoceptors using a combination of phenoxybenzamine (3 mg/kg i.v.), prazosin (0.1 mg/kg i.v.) and rauwolscine (1 mg/kg i.v.), higher doses of ibopamine, epinine and dopamine produced a propranolol-sensitive, beta-1 adrenoceptor-mediated positive chronotropic response that was significantly reduced in reserpine-pretreated rats, indicating a significant indirect component in the activity of these compounds at the level of the myocardium. Epinine and dopamine were equipotent and were 10 times more potent than ibopamine as directly acting beta-1 adrenoceptor agonists.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3039116 TI - Potentiation of the protective effects of a converting enzyme inhibitor and a thromboxane synthetase inhibitor in hemorrhagic shock. AB - The effect of a specific inhibitor of thromboxane (Tx) A2 synthesis, CGS-13080, a new angiotensin converting enzyme inhibitor, CGS-16617, and a combination of both drugs was studied in hemorrhagic shock in rats. Treatment with CGS-16617 (1 microgram/kg) or CGS-13080 (200 micrograms/kg) alone did not alter significantly postoligemic hypotension or the increase in plasma cathepsin D activity in shocked rats, compared with hemorrhaged rats receiving only their vehicle. Combined treatment with both drugs maintained postreinfusion mean arterial blood pressure and attenuated the increase in plasma cathepsin D activity in hemorrhaged rats. Treatment of shocked rats with each drug alone attenuated the accumulation of a myocardial depressant factor activity in the plasma, but the lowest myocardial depressant factor activities were observed in rats treated with the drug combination. Additionally, animals treated with the drug combination exhibited significantly longer postreinfusion survival times than rats receiving either the vehicle (P less than .01), CGS-16617 (P less than .05) or CGS-13080 (P less than .02). CGS-16617 (1 microgram/kg) attenuated significantly the pressor response to angiotensin I throughout the shock period. CGS-13080 attenuated the increase in TxB2 plasma concentrations in shock when compared with hemorrhaged rats receiving the vehicle (P less than .05). Greater attenuation of TxB2 was found after treatment with the drug combination (P less than .01 from vehicle, P less than .05 from CGS-13080 alone). CGS-16617, but not CGS-13080, was also found to have a direct antiproteolytic action in pancreatic homogenates. However, the drug combination (CGS-16617 and CGS-13080) decreased proteolytic activity even further (P less than .001) from CGS-16617 alone.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3039118 TI - Inhibition of adrenergic neurotransmission in canine tibial artery after exposure to 5-hydroxytryptamine in vitro. AB - Blood vessels may be exposed to 5-hydroxytryptamine when platelets aggregate and release vasoactive substances at sites of damage or disease. The functional consequences were studied of exposing the dog tibial artery for 2 hr to 5 hydroxytryptamine (10(-6) M) in vitro. During the exposure, unmetabolized 5 hydroxytryptamine was accumulated by the cocaine-sensitive amine uptake mechanism of tibial artery adrenergic nerves. After exposure to [3H]-5-hydroxytryptamine, transmural electrical stimulation caused the release of the tritiated indoleamine which was blocked by tetrodotoxin. After a 1-hr washout of rings of tibial artery exposed previously to 5-hydroxytryptamine, contractions in response to transmural electrical stimulation were depressed, whereas the response to exogenously added norepinephrine was unaffected. That the decreased response to electrical stimulation after exposure to 5-hydroxytryptamine was due to decreased release of norepinephrine from adrenergic nerves was demonstrated in strips of the artery preincubated in [3H]norepinephrine. The inhibition of [3H]norepinephrine release after exposure to 5-hydroxytryptamine, was blocked by the serotonergic antagonist, methiothepin, but not by the alpha adrenergic antagonist, phentolamine, suggesting that serotonergic receptors mediate the inhibition. The inhibition of [3H]norepinephrine release also was prevented by blocking adrenergic neuronal uptake with cocaine before exposure to 5-hydroxytryptamine. These results suggest that 5-hydroxytryptamine is accumulated and released by tibial artery adrenergic nerves as a cotransmitter. In so doing, the indoleamine inhibits adrenergic neurotransmission in the tibial artery by its action at prejunctional serotonergic receptors. PMID- 3039119 TI - Neurokinin-induced salivation in the anesthetized rat: a three receptor hypothesis. AB - Substance P (3 micrograms/kg), neurokinin A (20 micrograms/kg), neurokinin B (6 micrograms/kg) and acetylcholine (875 micrograms/kg) all produced salivation upon i.v. infusion in the anesthetized rat. Against single equivalent agonist doses, atropine (135 micrograms/kg i.v.) blocked both acetylcholine- and neurokinin B-, but not substance P- or neurokinin A-induced salivation. [D-Pro2,D-Trp7,9] substance P (1 mg/kg i.v.), a putative substance P antagonist, reduced responses to mammalian neurokinins but caused a 2-fold potentiation of acetylcholine induced salivation. [D-Pro2,D-Trp6,8,Nle10]-Neurokinin B (1 mg/kg i.v.), a novel putative neurokinin B antagonist, significantly reduced substance P- and neurokinin B- but not acetylcholine- or neurokinin A-induced salivation. The three agonists (at doses that produced salivation) and [D-Pro2,D-Trp6,8,Nle10] neurokinin B (1 mg/kg i.v.) lowered blood pressure in anesthetized rats by 35 to 40%. [D-Pro2,D-Trp7,9]-Substance P (1 mg/kg i.v.) had no significant effect on blood pressure. Hydralazine at 0.60 mg/kg (i.v.), a dose which lowered blood pressure by 47%, did not reduce substance P-induced salivation. Thus, blockade of neurokinin-induced salivation by [D-Pro2,D-Trp6,8,Nle10]-neurokinin B was probably not due to hypotension. Based on the differential effects of the three antagonists on neurokinin- and acetylcholine-induced salivation, we hypothesize the existence of three distinct neurokinin receptors in rat salivary gland, and suggest that neurokinin B receptors reside presynaptically. PMID- 3039120 TI - Relative efficacies of piperazines at the phosphoinositide hydrolysis-linked serotonergic (5-HT-2 and 5-HT-1c) receptors. AB - Serotonin (5-HT)-stimulated phosphoinositide hydrolysis is mediated by the 5-HT-2 receptor in rat cerebral cortex and by the 5-HT-1c receptor in rat choroid plexus. These systems were used to determine relative efficacies of piperazine derivatives at the 5-HT-2 and 5-HT-1c receptors. Both quipazine and 6-chloro-2-[1 piperazinyl]-pyrazine (MK-212) stimulated phosphoinositide hydrolysis in cerebral cortex, and these effects were blocked by ketanserin. The maximum responses to these agonists were 80% of the maximum response to 5-HT. m Trifluoromethylphenylpiperazine (TFMPP), m-chlorophenylpiperazine (MCPP) and 1-(1 naphthyl)-piperazine (1-NP) did not stimulate phosphoinositide hydrolysis in cerebral cortex at concentrations that blocked the effect of 5-HT. In the choroid plexus, TFMPP and MCPP, as well as MK-212 and quipazine, increased phosphoinositide hydrolysis and mianserin blocked these effects. MK-212 had an efficacy which was equal to that of 5-HT, whereas quipazine, MCPP and TFMPP were partial agonists in the choroid plexus. 1-NP did not stimulate phosphoinositide hydrolysis in choroid plexus but completely blocked the effect 5-HT. On the basis of these data, we conclude that quipazine and MK-212 are partial agonists at 5-HT 2 receptors in cerebral cortex, whereas 1-NP, TFMPP and MCPP are pure antagonists of the cortical 5-HT-2 receptor. However, TFMPP and MCPP as well as quipazine and MK-212 are agonists at the 5-HT-1c receptor, while 1-NP is a pure antagonist of the 5-HT-1c receptor in choroid plexus. PMID- 3039121 TI - Beta-endorphin stimulates human polymorphonuclear leukocyte superoxide production via a stereoselective opiate receptor. AB - Opioid peptides have been shown to modulate the function of cells associated with host defense. Both opiate and nonopiate receptor mechanisms have been shown to mediate cell responses to these peptides. In this study we used a ferricytochrome C reduction microassay to measure superoxide (O2-) production by human polymorphonuclear leukocytes after stimulation with beta-endorphin (beta-END). beta-END was found to stimulate O2- release at concentrations from 10(-14) to 10( 8) M; the peak response occurred at 10(-12) M. A microassay based on the horseradish peroxidase-mediated oxidation of phenol red was used to demonstrate the production of hydrogen peroxide H2O2, by beta-END at 10(-12) M. The accumulation of H2O2 was reduced by the inhibitor, nitroprusside, and by the converting enzyme, catalase. The accumulation of O2- in response to the potent chemotactic peptide formyl-methionine-leucine-phenylalanine was studied and a distinctly different dose-response profile with a peak response at 10(-8) M was observed. Because beta-END can apparently bind to and activate cellular functions by nonopiate receptors, N-acetyl-beta-END was tested. At doses between 10(-14) and 10(-8) M, it failed to effect O2- accumulation. Moreover, (-)-naloxone 10( 12) M was shown to completely abolish the stimulatory effect of equimolar beta END whereas (+)-naloxone was entirely ineffective. At 10(-8) M both stereoisomers also failed to inhibit formyl-methionine-leucine-phenylalanine 10(-8) M. Thus, at the picomolar concentration present in the human systemic circulation, beta-END activates oxygen metabolism by polymorphonuclear leukocytes through stereoselective, naloxone-sensitive opiate receptors. PMID- 3039122 TI - Dextrorphan and levorphanol selectively block N-methyl-D-aspartate receptor mediated neurotoxicity on cortical neurons. AB - Neocortical neurons in cell cultures prepared from fetal mice were impaled for intracellular recording. Dextrorphan (DX), a clinically tested dextrorotatory morphinan lacking opioid action, did not alter neuronal membrane potential or conductance, but produced a selective attenuation of N-methyl-D-aspartate (NMDA) responses; kainate and quisqualate responses were not affected. DX also antagonized morphological and chemical (lactate dehydrogenase efflux) evidence of cortical neuronal cell injury produced by toxic bath exposure to NMDA, quinolinate or glutamate, but did not affect toxic exposure to quisqualate or kainate. This selective antagonism of neurotoxicity was apparent at micromolar concentrations of DX with an ED50 of 13 to 17 microM. A similar, but less potent neuron-protective effect, was seen with the opioid levorotatory enantiomer of DX, levorphanol (ED50, 40 microM). The O-methyl derivative of DX, dextromethorphan, also antagonized NMDA and glutamate neurotoxicity, but with possibly lower efficacy than DX. The higher potency of DX over levorphanol suggests that this novel neuron-protective action is not mediated by classic opiate receptors; it may be mediated at the "sigma opiate"/phencyclidine site. If further studies establish that DX and related compounds retain neuron-protective efficacy in appropriate animal models, the established clinical safety record of DX and dextromethorphan may allow prompt investigation of the NMDA receptor-blockade strategy in certain neurological disease states. PMID- 3039123 TI - The differential effects of twitch potentiators on charge movements in frog skeletal muscle. AB - The effects of twitch potentiators at physiologically effective concentrations on intramembrane charge movements were examined in voltage-clamped frog skeletal muscle in the presence and absence of tetracaine. Caffeine, even at high concentrations (2.5 mM) did not alter the potential dependence of either tetracaine-sensitive or tetracaine-resistant portions of non-linear charge through a voltage range that included the mechanical threshold. Perchlorate (8 mM) altered the form and shifted the potential dependence of the non-linear charge by about -25 mV in the hyperpolarizing direction, but did not alter the total available charge. Comparing charge movements in the presence and absence of 4 mM-tetracaine demonstrated that perchlorate shifted the threshold and voltage dependence of the delayed ('q gamma') transients but had no action on the steady state tetracaine-resistant charge. Thiocyanate (10 mM) shifted the voltage dependence of charge movements by about 20 mV in the hyperpolarizing direction. Experiments employing 4 mM-tetracaine demonstrated that this was the result of effects on both tetracaine-sensitive and tetracaine-resistant charge movements. On the basis of their effects on contractile activation reported on earlier occasions, these differential effects of twitch potentiators are explicable in terms of tetracaine-sensitive and tetracaine-resistant charge components being causally separate and not requiring a direct participation of tetracaine resistant charge in contractile activation. PMID- 3039124 TI - Quantal analysis of action of hemicholinium-3 studied at a central cholinergic synapse of Aplysia. AB - The effects of hemicholinium-3 (HC-3) on cholinergic transmission were studied on central identified inhibitory (H-type post-synaptic cell, Cl- channels) and on excitatory (D-type post-synaptic cell, cationic channels) synapses of Aplysia californica. In the H-type post-synaptic cell, the amplitude and the decay time of miniature post-synaptic currents (m.p.s.c.s.) were calculated by statistical analysis of long duration induced post-synaptic current (l.d.i.p.s.c.) due to 3 s depolarizations of the presynaptic neurone in the presence of tetrodotoxin. On H type receptors, with respect to acetylcholine (ACh), HC-3 acted as an agonist and a blocker whereas on D-type receptors, it acted only as a blocker. At low concentration of bath-applied HC-3, in the H-type synapse, the decay time of the evoked inhibitory post-synaptic current (i.p.s.c.) as well as that of the m.p.s.c. was lengthened. These changes were rapidly reversible by wash. The decay time of excitatory post-synaptic current (e.p.s.c.) at the D-type synapse was not affected. On the inhibitory synapse, HC-3 applied in the bath at the concentration of 10(-5) M, reduced considerably the size of the m.p.s.c.s whereas the evoked i.p.s.c.s and the l.d.i.p.s.c.s were only slightly affected pointing to an increase of the quantal content of both responses. After wash, both i.p.s.c.s and l.d.i.p.s.c.s showed a clear facilitation which persisted for several tens of minutes. The presence of presynaptic receptors was considered. Similar facilitation of e.p.s.c.s by HC-3 was observed at the D-type synapse. The comparison of the degree of depression by HC-3 of the m.p.s.c.s and of the responses to ionophoretically applied ACh, indicated that the size of the quantum was not changed. Intracellular injection of HC-3 into the presynaptic neurone of the H-type synapse led to a decrease of transmitter release which affected solely the quantal content of the responses. As the synaptic transmission could not be restored by injection of exogenous ACh into the presynaptic neurone, it was concluded that the depression of transmission was not due to a decrease of ACh synthesis. PMID- 3039126 TI - Study of Coxsackie B viral infections in chronic pancreatitis patients from Kerala. PMID- 3039127 TI - Pleomorphic adenoma of the lacrimal gland (a case report). PMID- 3039125 TI - Muscarinic slow excitation and muscarinic inhibition of synaptic transmission in the rat neostriatum. AB - Intracellular recording from neurones in rat neostriatal slices was used to compare the muscarinic effects of endogenous acetylcholine (ACh) released from cholinergic neostriatal synapses with the action of exogenously applied muscarinic agonists. Repetitive electrical stimulation in the neostriatum evoked a series of fast excitatory post-synaptic potentials (e.p.s.p.s) followed by a short, variable period of input resistance decrease. In the presence of the acetylcholinesterase (AChE) inhibitor, physostigmine, these potentials were followed by a slow e.p.s.p. which lasted about 60 s. Higher stimulus intensities were needed to elicit the slow e.p.s.p. than the fast e.p.s.p. The slow e.p.s.p. could not be observed after a single stimulus. Its amplitude was graded and increased with stimulus strength. The slow e.p.s.p. was blocked by the muscarinic antagonist atropine (10 microM) and by Ba2+ (100 microM). Input resistance increased during the slow e.p.s.p. Depolarization of the cell increased the size of the slow e.p.s.p. and hyperpolarization decreased it. Simultaneously, resting input resistance increased with membrane depolarization and decreased with membrane hyperpolarization. Repetitive intrastriatal stimulation was followed by a hyperpolarization instead of the depolarization at membrane potentials negative to -75 mV. Input resistance increased during this hyperpolarization as it did during the slow e.p.s.p. The slow e.p.s.p. persisted at membrane potentials of 70 to -80 mV if K+ concentration in the saline was reduced from 5 to 2 mM. In 10 mM-K+, the repetitive stimulation was followed by a hyperpolarization even at membrane potentials as low as -60 to -50 mV. Bath perfusion of high concentrations (100 microM) of muscarine or carbachol induced a sustained increase in the input resistance. The muscarinic agonists also reduced the amplitude of intrastriatally evoked fast e.p.s.p.s; however, this effect was transient and compensated by the increase in input resistance. The effects of the muscarinic agonists on input resistance and e.p.s.p. amplitude were antagonized by atropine (10 microM). Sustained decreases of e.p.s.p. amplitude were induced by the bath application of low doses (0.5-10 microM) of muscarine or carbachol. Input resistance was not altered. Atropine (1-10 microM) antagonized this effect. A sustained reduction of fast e.p.s.p. amplitude resulted also from inhibition of AChE by application of physostigmine (1-100 microM). Input resistance and neuronal excitability were not affected by AChE blockade.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3039129 TI - Antibodies to topoisomerase I in sera from patients with scleroderma. AB - In a study of sera from patients with scleroderma, it was found that Scl-70 is the abundant nuclear enzyme DNA topoisomerase I. PMID- 3039128 TI - GABA-mediated synaptic inhibition of projection neurons in the antennal lobes of the sphinx moth, Manduca sexta. AB - Responses of neurons in the antennal lobe (AL) of the moth Manduca sexta to stimulation of the ipsilateral antenna by odors consist of excitatory and inhibitory synaptic potentials. Stimulation of primary afferent fibers by electrical shock of the antennal nerve causes a characteristic IPSP-EPSP synaptic response in AL projection neurons. The IPSP in projection neurons reverses below the resting potential, is sensitive to changes in external and internal chloride concentration, and thus is apparently mediated by an increase in chloride conductance. The IPSP is reversibly blocked by 100 microM picrotoxin or bicuculline. Many AL neurons respond to application of GABA with a strong hyperpolarization and an inhibition of spontaneous spiking activity. GABA responses are associated with an increase in neuronal input conductance and a reversal potential below the resting potential. Application of GABA blocks inhibitory synaptic inputs and reduces or blocks excitatory inputs. EPSPs can be protected from depression by application of GABA. Muscimol, a GABA analog that mimics GABA responses at GABAA receptors but not at GABAB receptors in the vertebrate CNS, inhibits many AL neurons in the moth. PMID- 3039130 TI - Maintenance therapy with unprocessed bran in the prevention of acute anal fissure recurrence. AB - The effect of unprocessed bran in a dose of 5 g three times daily and a dose of 2.5 g three times daily for one year on the recurrence rate of anal fissures was studied in a double-blind, placebo-controlled trial in 90 patients with recently healed acute posterior anal fissures. Fifteen patients (16.6%) were withdrawn before the code was broken due to failure to follow the trial protocol for various reasons. Significantly fewer recurrences occurred in patients receiving bran 5 g three times daily (recurrence rate 16%, 95% confidence limits, 4.54 to 36.08) when compared with patients receiving bran 2.5 g three times daily (60%; 38.67 to 78.87) (P less than 0.01) and with patients receiving placebo three times daily (68%; 46.50 to 85.05) (P less than 0.01). No significant difference in recurrences was found between patients on bran 2.5 g and those on placebo. PMID- 3039131 TI - Cardiotonic agents. 1. Novel 8-aryl-substituted imidazo[1,2-a]- and -[1,5 a]pyridines and imidazo[1,5-a]pyridinones as potential positive inotropic agents. AB - Several 8-arylimidazo[1,2-a]pyridines, 8-arylimidazo[1,5-a]pyridines, and 8 arylimidazo[1,5-a]pyridinones were prepared and tested in vitro for potential cardiac inotropic and electrophysiological activity. Selected analogues were further tested in vivo in canine hemodynamic and cardiac electrophysiology models. Compounds having an imidazole substituent consistently showed activity. A pharmacophoric relationship between heterocycle-phenyl-imidazole and positive inotropic activity was noted. The significance of this relationship is discussed. PMID- 3039132 TI - Cardiotonic agents. 2. (Imidazolyl)aroylimidazolones, highly potent and selective positive inotropic agents. AB - A series of 4-alkyl-1,3-dihydro-5-[(1H-imidazolyl)benzoyl]-2H-imidazol-2-ones 9 was synthesized and evaluated in vitro for positive inotropic and cyclic AMP phosphodiesterase inhibitory activity. A wide range of inotropic and enzyme inhibitory potencies was observed, substitution on the imidazolyl moiety being the major determinant of activity. The 4-ethyl-5-[4-(1H-imidazol-1-yl)benzoyl] congener 9g exhibited the highest potency in vitro. Incorporation of a methyl group at the imidazolyl 2-position gave 9h, which was less potent but remarkably selective in vivo for positive inotropic effects over heart rate and hypotensive effects. PMID- 3039133 TI - Tricyclic compounds as selective antimuscarinics. 1. Structural requirements for selectivity toward the muscarinic acetylcholine receptor in a series of pirenzepine and imipramine analogues. AB - The M1-selective antiulcer drug pirenzepine (1) is a tricyclic compound with close resemblance to tricyclic psychotropic agents such as imipramine (2). Despite this fact, pirenzepine is devoid of any psychotropic effects, exhibiting measurable antagonistic effects in biochemical assays and receptor binding studies only toward the muscarinic receptor system. To understand how different groups in these tricyclic molecules affect binding affinities, a set of nine compounds structurally related to pirenzepine (1) and imipramine (2) has been selected for analysis, comprising three different tricycles and three different side chains. The compounds were tested for their affinity to the imipramine and muscarinic receptors in homogenized rat cortex tissue. The result of these studies suggests that it is the nature and placement of accessory groups that determine the differences in receptor recognition and the binding process. In the case of pirenzepine (1), preferential binding toward the muscarinic receptor is brought about by the endocyclic amide group, by the positioning of the protonated N atom of the side chain, and to a minor extent by the exocyclic amide group. From these findings a putative model for the explanation of selective binding of pirenzepine (1) to the muscarinic receptor has been derived. PMID- 3039134 TI - Synthesis of two glucagon antagonists: receptor binding, adenylate cyclase, and effects on blood plasma glucose levels. AB - In diabetes mellitus, hyperglycemia is often associated with elevated levels of glucagon in the blood. This suggests that glucagon is a contributing factor in the metabolic abnormalities of diabetes mellitus. A glucagon-receptor antagonist would provide direct evidence for glucagon's role in diabetes mellitus. On the basis of careful consideration of conformational, amphiphilic, and structural factors, we have synthesized two new glucagon analogues with antagonist biological activities by using solid-phase methodology. These two new analogues, [Asp3,D-Phe4,Ser5,Lys17,18,Glu21]glucagon (2) and [D-Phe4,Tyr5,3,5-I2 Tyr10,Arg12,Lys17,18,G lu21]glucagon (3) had IC50 values 5.4% and 50% those of glucagon, respectively, and showed no measurable adenylate cyclase activity. When tested in normal rats, 2 lowered plasma glucose levels and suppressed glucagon mediated hyperglycemia 105 +/- 8%, back to basal levels. Analogue 3, which lowered the basal adenylate cyclase activity in rat liver plasma membranes, increased plasma glucose levels at very high concentration in vivo and inhibited glucagon-mediated hyperglycemia in normal rats by 50%. However, neither of the new glucagon antagonists lowered the plasma glucose levels of diabetic animals. The data would suggest these new glucagon-receptor antagonists may have two actions: (a) in normal rats they can act as standard glucagon-receptor inhibitors of glucagon-mediated glycogenolysis; (b) in diabetic rats, however, because of the low levels of glycogen in the liver, the antagonists apparently have little or no antagonist effect or enhancement on glucagon-mediated glucose production. PMID- 3039135 TI - Synthesis and cardiotonic activity of a series of substituted 4-alkyl-2(1H) quinazolinones. AB - The synthesis, cardiac fraction III cyclic nucleotide phosphodiesterase (PDE-III) inhibition, and positive inotropic activity of a series of 2(1H)-quinazolinones are reported. A general synthesis of the series involved the cyclization of 2 aminoacetophenones with potassium cyanate in acetic acid. Modifications at the 4 position of the quinazoline nucleus were best achieved by formation of the intermediate N1-acyl-N3-phenylurea from the substituted phenyl isocyanate and appropriate carboxamide. PPA was used to ring close to the quinazoline product. Generally the SAR for the series paralleled the five-point model previously published for PDE-III inhibition. The most active analogue of the series was 5,6 dimethoxy-4-methyl-2(1H)-quinazolinone (1) (ORF 16600), which had about twice the intravenous potency of amrinone. Compound 1 is currently under development as an orally active cardiotonic. PMID- 3039136 TI - Pyrroloisoquinoline antidepressants. 2. In-depth exploration of structure activity relationships. AB - A series of pyrrolo[2,1-a]isoquinolines, and related compounds, were examined for antidepressant-like activity, by virtue of their antagonism of tetrabenazine induced ptosis and sedation, and inhibition of biogenic amine uptake. Thus, we have identified some of the most potent antagonists of TBZ-induced ptosis and some of the most potent inhibitors of the uptake of dopamine, norepinephrine, and serotonin (in rat brain synaptosomes) ever reported. Compounds of particular note, in this regard, are 52b, 29b, 22b, and 48b, respectively. Biological activity was chiefly manifested by the trans isomeric class. Also, through resolution of four compounds, 7b, 24b, 37b, and 48b, biological activity was found to be associated with the (+) enantiomer subgroup (salts measured at 589 nm in MeOH), corresponding to the 6S, 10bR absolute configuration for 7b, 37b, and 48b, and the 6R,10bR configuration for 24b. An X-ray determination on (+)-24b X HBr established its absolute configuration; configurations for the other compounds were verified by enantiospecific synthesis starting with (+)-(R)-2 phenylpyrrolidine. Regarding the pendant phenyl ring, diverse substitution patterns were investigated. Those substitutions that were particularly unfavorable were 3',4',5'-trimethoxy (20b), 2',3',4',5',6'-pentafluoro (34b), 2' trifluoromethyl (38b), 3',5'-bis(trifluoromethyl) (42b), 4'-n-butyl (44b), 2' cyano (47b), 4'-methylsulfonyl (50b), and 2'-carboxy (58b). Exceedingly potent compounds, in one way or another, were 10b-12b, 22b, 23b, 25b, 28b, 29b, 33b, 45b, 48b, 51b-53b. The pattern of aromatic substitution had a strong impact on selectivity in the uptake tests (NE vs. DA vs. 5-HT). Activity was significantly diminished by methyl substitution of 7b at the 5 (65, 66), 6 (61b), or 10b (60b) position, by changing the phenyl group of 7b to cyclohexyl (67b), benzyl (68b), or H (72), by moving the phenyl group of 7b to the 5 (69) or 10b (70) position, by expansion of ring B to an azepine (78b), and by modification of ring C to an azetidine (77b), piperidine (75b), or azepine (74b). The interaction of selected analogues with various CNS receptors is reported. Little affinity was shown for the muscarinic cholinergic receptor, suggesting a lack of anticholinergic side effects. Interestingly, 24b and 33b displayed a high affinity for the serotonin-2 receptor, analogous to mianserin and clomipramine. After the body of data was reviewed, derivatives 24b and 48b were chosen for advanced development. PMID- 3039137 TI - Synthesis and beta-lactamase inhibitory properties of 2 beta-[(1,2,3-triazol-1 yl)methyl]-2 alpha-methylpenam-3 alpha-carboxylic acid 1,1-dioxide and related triazolyl derivatives. AB - Benzhydryl 2 beta-[(1,2,3-triazol-1-yl)methyl]-2 alpha-methylpenam- 3 alpha carboxylate 1,1-dioxide was prepared by heating benzhydryl 2 beta-(azidomethyl)-2 alpha-methylpenam-3 alpha-carboxylate 1,1-dioxide with (trimethylsilyl)acetylene. The ester group was removed by hydrogenolysis to give sodium 2 beta-[(1,2,3 triazol-1-yl)methyl]-2 alpha-methylpenam-3 alpha-carboxylate 1,1-dioxide (3i, YTR 830), which was found to be a potent inhibitor of various bacterial beta lactamases. A series of related compounds was prepared in a similar way, and all of these compounds show excellent beta-lactamase inhibitory properties. PMID- 3039138 TI - Synthesis and antidepressant properties of novel 2-substituted 4,5-dihydro-1H imidazole derivatives. AB - A unique combination of alpha 2-adrenoreceptor antagonist and serotonin-selective reuptake inhibitory activities has been identified in a series of 2-substituted 4,5-dihydro-1H-imidazole derivatives. This combination of blocking activities has provided one of these derivatives, napamezole hydrochloride (2), with potential as an antidepressant. A discussion of the syntheses of these compounds includes a convenient method for the conversion of nitriles to imidazolines with ethylenediamine and trimethylaluminum. PMID- 3039139 TI - X-ray crystal structure of the opioid ligand naltrexonazine. AB - The anti-anti isomer of naltrexonazine (1) was synthesized, and its configuration was confirmed by X-ray crystallography. The syn-anti isomer is readily converted to 1 under acidic conditions. The apparent equal receptor binding of 1 and syn anti isomer indicates that isomerization of the azine moiety may take place under the conditions of biological evaluation. Two possible explanations for wash resistant binding of 1 to opioid receptors are presented. The first possibility involves a noncovalent interaction of the ligand with the opioid receptor, and the second considers covalent binding by a receptor-based sulfhydryl group. PMID- 3039140 TI - Synthesis and biological activity of 5-(2,2-difluorovinyl)-2'-deoxyuridine. AB - 5-(2,2-Difluorovinyl)uracil (IV) was synthesized from 2,4-dimethoxy-5 bromopyrimidine by sequential formylation, difluoromethylenation, and removal of the 2- and 4-methyl groups. Condensation of the trimethylsilyl derivative of IV with protected D-erythro-pentofuranosyl chloride followed by separation of anomers and deblocking gave 5-(2,2-difluorovinyl)-2'-deoxyuridine (V). Compound V was active against herpes simplex virus type 1 (HSV-1) infection as well as tumor cells transformed by the HSV-1 thymidine kinase gene. PMID- 3039141 TI - Hypoplastic tibiae with postaxial polysyndactyly: a new dominant syndrome? AB - A five year old boy is reported with hypoplastic, bowed tibiae and postaxial polysyndactyly. Sixteen relatives of both sexes in four generations either have bilateral syndactyly or postaxial polydactyly or both. The nature of the condition and the possible mode of inheritance are discussed. PMID- 3039142 TI - Does a relationship exist between neutrophil myeloperoxidase deficiency and the occurrence of neoplasms? AB - 36 unrelated individuals with neutrophil MPO deficiency, (10 totally MPO deficient) were found on screening a population of 148,000 subjects. A further 2 subjects with total and 22 with partial MPO deficiency were identified through family studies. The assessment of neutrophil function, i.e., peroxidase activity, superoxide anion generation, microbicidal activity towards fungi and bacteria, and locomotor behaviour, was carried out in 10 subjects with total and 4 with partial MPO deficiency. We found that the enzyme defect is associated with a marked impairment in the killing of both S. aureus and C. albicans, without affecting microbicidal activity against S. faecalis. There appears to be a high incidence of malignancy in patients with complete MPO deficiency, suggesting a relationship between a defective MPO system and neutrophil-mediated tumor cell cytotoxicity. PMID- 3039143 TI - Evaluation of serum parameters relevant to vitamin D status in tamarins. AB - Serum levels of 25(OH)D, alkaline phosphatase (AP), and parathormone (PTH) were evaluated to investigate the vitamin D requirement of Saguinus fuscicollis. Diets with various vitamin D content were fed 4 weeks and longer. The values of 25(OH)D (30-300 nmol/l), AP (less than 300 U/l), and PTH (less than or equal to 1,000 equl/l) considered as normal were obtained with 2,000 IU D3/kg diet, or 33 IU/animal/day, which we regard as the level required. Animals depleted of vitamin D for 215 days developed a secondary hyperparathyroidism. PMID- 3039144 TI - D2O and the sodium pump in squid nerve membrane. AB - In 10 K artificial seawater (ASW), D2O replacement reduced the Na efflux of squid axons by about one third. In 0 K ASW, D2O replacement had little effect. D2O reduced the K+ sensitivity of the efflux but increased the affinity for K+. A 4 degrees decrease in temperature mimicked the effects of D2O. When axons were injected with arginine, to decrease the ATP/ADP ratio, they lost K+ sensitivity in normal ASW, as expected. Their efflux into 0 K ASW became D2O sensitive. The results are discussed in terms of conformational changes in the Na pump molecular complex. PMID- 3039146 TI - Nucleotide sequence and comparative analysis of the human papillomavirus type 18 genome. Phylogeny of papillomaviruses and repeated structure of the E6 and E7 gene products. AB - The complete nucleotide sequence and genomic organization of human papillomavirus type 18, associated with cervical cancer, has been established. A detailed comparative analysis was undertaken leading to the identification of a number of features specific for genital papillomaviruses and the construction of a phylogenetic tree. Genital papillomaviruses differ from other human and animal papillomaviruses as they possess a longer E1 open reading frame (ORF) and have a characteristic control region. Phylogenetically, HPV 18 is located between the benign genital viruses, HPV 6 and HPV 11, and the malignant isolates, HPV 16 and HPV 33, and may represent an evolutionary intermediate among oncogenic papillomaviruses. Viral gene products known to be involved in cellular transformation are those of ORFs E5, E6 and E7. Significant sequence variation was found between the E6 to E7 regions of different integrated forms of HPV 18. On re-examination of the E6 primary structures we noticed that the gene has evolved by successive duplications of a unit encoding 33 amino acids, which include a Cys-X-X-Cys motif. Furthermore, the E7 gene product has apparently evolved in the same manner and is related to E6. Both gene products bear a striking resemblance to the transcriptional factor IIIA of Xenopus laevis, the prototype of a new class of nucleic acid binding proteins. PMID- 3039145 TI - Measurements of intracellular pH in single LLC-PK1 cells: recovery from an acid load via basolateral Na+/H+ exchange. AB - LLC-PK1 cells (a continuous epithelioid cell line with renal characteristics) are examined by microspectrofluorometry as single cells, in order to determine the mechanism of intracellular pH (pHi) recovery from an acid load imposed by ammonium preincubation and removal (NH4 prepulse). Initial experiments evaluate the intracellular K+ levels through a null point analysis of total cellular K+ with flame photometry. The response of BCECF (a pH-sensitive fluorescent dye) is then calibrated, using saturating concentrations of nigericin to cause defined changes in pH. For experiments with the microspectrofluorometer, LLC-PK1 cells were grown on either glass coverslips or filters (the latter attached to plastic coverslips with a hole under the filter). The cells on glass coverslips demonstrate a Na+-dependent recovery from an (NH4 prepulse) acid load which is sensitive to 1 microM ethylisopropylamiloride. They also demonstrate a 'set point' of activation of Na+/H+ exchange. When examined for changes in pH, due to changes in membrane potential, plasma membrane proton conductance could not be detected at resting pHi. Cells grown on filters also demonstrate a pHi recovery from an acid load which is Na+ dependent and ethylisopropylamiloride sensitive, but in this configuration, the majority of cells (22/23 preparations) require Na+ at the basolateral membrane for rapid pHi recovery. The morphology and polarity of the cells grown on permeable supports appears normal at the electron microscopic level. The results are not affected by changes in cell seeding density or collagen treatment of the filters. PMID- 3039147 TI - Frameshift suppressor mutations affecting the major glycine transfer RNAs of Saccharomyces cerevisiae. AB - Mutations have been identified in Saccharomyces cerevisiae glycine tRNA genes that result in suppression of +1 frameshift mutations in glycine codons. Wild type and suppressor alleles of genes encoding the two major glycine tRNAs, tRNA(GCC) and tRNA(UCC), were examined in this study. The genes were identified by genetic complementation and by hybridization to a yeast genomic library using purified tRNA probes. tRNA(UCC) is encoded by three genes, whereas approximately 15 genes encode tRNA(GCC). The frameshift suppressor genes suf1+, suf4+ and suf6+ were shown to encode the wild-type tRNA(UCC) tRNA. The suf1+ and suf4+ genes were identical in DNA sequence, whereas the suf6+ gene, whose DNA sequence was not determined, was shown by a hybridization experiment to encode tRNA(UCC). The ultraviolet light-induced SU F1-1 and spontaneous SU F4-1 suppressor mutations were each shown to differ from wild-type at two positions in the anticodon, including a +1 base-pair insertion and a base-pair substitution. These changes resulted in a CCCC four-base anticodon rather than the CCU three-base anticodon found in wild-type. The RNA sequence of tRNA(UCC) was shown to contain a modified uridine in the wobble position. Mutant tRNA(CCCC) isolated from a SU F1-1 strain lacked this modification. Three unlinked genes that encode wild-type tRNA(GCC), suf20+, trn2, and suf17+, were identical in DNA sequence to the previously described suf16+ frameshift suppressor gene. Spontaneous suppressor mutations at the SU F20 and SU F17 loci were analyzed. The SU F20-2 suppressor allele contained a CCCC anticodon. This allele was derived in two serial selections through two independent mutational events, a +1 base insertion and a base substitution in the anticodon. Presumably, the original suppressor allele, SU F20 1, contained the single base insertion. The SU F17-1 suppressor allele also contained a CCCC anticodon resulting from two mutations, a +1 insertion and a base substitution. However, this allele contained an additional base substitution at position 33 adjacent to the 5' side of the four-base anticodon. The possible origin and significance of multiple mutations leading to frameshift suppression is discussed. PMID- 3039148 TI - Cascade regulation of Caulobacter flagellar and chemotaxis genes. AB - The assembly of a functional flagellum in the bacterium Caulobacter crescentus requires the protein products of approximately 30 genes expressed in a temporally discrete and spatially distinct manner. Our current understanding of this system has been limited by the fact that purified protein products are available for only about one-fifth of these genes. A genetically engineered transposon promoter probe, Tn5-VB32, containing a promoterless gene encoding neomycin phosphotransferase II (NPTase II) was used to generate a series of non-motile (fla-), kanamycin resistant strains of C. crescentus. These transcription-fusions allow the expression of NPTase II to be controlled by flagellar promoters, and thus questions of temporal regulation of flagellar genes can be addressed without the need to obtain purified protein products. The flagellar promoters accessed by Tn5-VB32 exhibited temporal regulation analogous to the known flagellar and chemotaxis gene products. The expression of NPTase II in these mutants is read from a chimeric mRNA that initiates in a chromosomal fla promoter and continues through the inserted NPTase II gene. Thus, temporal regulation is controlled by modulating either the initiation of transcription, or transcript turnover, at specific times in the cell cycle. Epistatic interactions between the genes accessed by the promoter probe and other flagellar loci were studied in double fla mutants generated by transducing the promoter-probe mutations into spontaneously derived second-site fla-mutant backgrounds. The synthesis of both natural fla gene products and the accessed NPTase II was assayed in these strains using antisera to purified components of the flagellum and to purified NPTase II. On the basis of these interactions, a trans-acting hierarchy of flagellar and chemotaxis gene expression is proposed. PMID- 3039149 TI - Identification of a gene cluster involved in flagellar basal body biogenesis in Caulobacter crescentus. AB - The bacterial flagellum is a complex structure composed of a transmembrane basal body, a hook, and a filament. In Caulobacter crescentus the biosynthesis and assembly of this structure is under temporal and spatial control. To help to define the order of assembly of the flagellar components and to identify the genes involved in the early steps of basal body construction, mutants defective in basal body formation have been analyzed. Mutants in the flaD flaB flaC gene cluster were found to be unable to assemble a complete basal body. The flaD BC motC region was cloned and the genes were localized by subcloning and complementation analysis. A series of Tn5 insertion mutations in the flaD BC region were mapped. Complementation analysis of the Tn5 insertion mutants indicated the existence of at least four transcriptional units in the region and identified the presence of two new genes designated flbN and flbO. Mutants in flbN, flaB, flaC and flbO were unable to assemble any basal body structure and are likely to be involved in the early steps of basal body formation. The flaD mutant, however, was found to contain a partially assembled basal body consisting of the rod and three hook-distal rings. All of the mutants in this cluster exhibited pleiotropic effects on the expression of other flagellar and chemotaxis functions, including the level of synthesis of flagellins, the hook protein and hook protein precursor, and the level of chemotaxis methylation. PMID- 3039150 TI - Use of site-specific recombination as a probe of DNA structure and metabolism in vivo. AB - We used site-specific recombination catalyzed by the bacteriophage lambda Int system to probe DNA structure and metabolism in vivo. In vitro, the complexity of catenated products was linearly proportional to substrate supercoil density. A system was developed that gave efficient, controlled Int recombination in Escherichia coli cells. From a comparison of the data obtained in vitro and in vivo, we conclude that Int recombination does have the same mechanism in vivo as it has in vitro, but that only 40% of the plasmid DNA linking deficit in E. coli cells may be in the interwound supercoil form demonstrated in vitro. We suggest that this is the effective level of supercoiling in vivo, because the remaining DNA is constrained in alternative forms by protein binding. The study of Int recombination in vivo also provides an assay for enzymes that decatenate circular molecules, such as those formed during DNA replication. We find that DNA gyrase is the principal decatenase in E. coli and that it acts spontaneously and rapidly. PMID- 3039152 TI - Assignment of resonances in the 1H nuclear magnetic resonance spectrum of the carbon monoxide complex of human hemoglobin alpha-chains. AB - Assignments are reported for a substantial number of heme and amino acid proton resonances in the 1H nuclear magnetic resonance spectrum of the carbon monoxide complex of isolated hemoglobin alpha-chains. These resonances provide information on the solution conformation of the protein, particularly in the vicinity of the heme. The heme pocket structure is generally similar to that of carbonmonoxymyoglobin; several conserved residues adopt virtually identical positions relative to the heme in the two proteins. The largest conformational differences involve residues surrounding the ligand-binding site, notably Val62 (E11) and His58 (E7). The chemical shifts of the proximal His87 (F8) resonances are very similar in spectra of the two proteins, indicating a highly conserved coordination geometry and similar hydrogen bonding to the backbone carbonyl of Leu83 (F4). PMID- 3039151 TI - Sequence dependence of Drosophila topoisomerase II in plasmid relaxation and DNA binding. AB - The sequence dependence of Drosophila topoisomerase II supercoil relaxation and binding activities has been examined. The DNA substrates used in binding experiments were two fragments from Drosophila heat shock locus 87A7. One of these DNA fragments includes the coding region for the heat shock protein hsp70, and the other includes the intergenic non-coding region that separates two divergently transcribed copies of the hsp70 gene at the locus. The intergenic region was previously shown to have a much higher density of topoisomerase cleavage sites than the hsp70 coding region. Competition nitrocellulose filter binding assays demonstrate a preferential binding of the intergene fragment, and that binding specificity increases with increasing ionic strength. Dissociation kinetics indicate a greater kinetic stability of topoisomerase II complexes with the intergene DNA fragment. To study topoisomerase II relaxation activity, we used supercoiled plasmids that contained the same fragments from locus 87A7 cloned as inserts. The relative relaxation rates of the two plasmids were determined under several conditions of ionic strength, and when the plasmid substrates were included in separate reactions or when they were mixed in a single reaction. The relaxation properties of these two plasmids can be explained by a coincidence of high-affinity binding sites, strong cleavage sites, and sites used during the catalysis of strand passage events by topoisomerase II. Sequence dependence of topoisomerase II catalytic activity may therefore parallel the sequence dependence of DNA cleavage by this enzyme. PMID- 3039153 TI - Peripheral neuropathy in multiple sclerosis. AB - Evidence suggests that the spectrum of multiple sclerosis (MS) may extend to peripheral nerve dysfunction. The author describes a patient with MS whose peripheral nervous system was involved. PMID- 3039154 TI - An ultrastructural morphometric study of the effects of chronic melatonin administration on the zona fasciculata of rat adrenal cortex. AB - The effects of a long-term administration of melatonin on the adrenal zona fasciculata were investigated both in 'normal' rats and in animals whose hypothalamo-hypophyseal-adrenal axis had been interrupted by a prolonged infusion with dexamethasone or dexamethasone plus maintenance doses of ACTH. In the 'normal' rats, melatonin caused a notable atrophy of zona fasciculata cells, coupled with a significant lowering in the plasma concentration of corticosterone. On the contrary, in dexamethasone- or dexamethasone plus ACTH infused animals, melatonin induced an evident hypertrophy of zona fasciculata cells which, however, was not associated with a rise in the corticosterone blood level. The hypothesis is advanced that the direct stimulatory effect of melatonin on the growth of zona fasciculata in 'normal' rats can be masked by the concurrent inhibition of the hypophyseal release of ACTH. The possible mechanism of the direct effect of melatonin on the zona fasciculata is discussed in the light of evidence indicating that this hormone enhances the intracellular catabolism of corticosterone, which is well-known to exert a powerful direct depression of the growth and steroidogenic capacity of rat adrenocortical cells. PMID- 3039155 TI - The effect of OH(.) radicals generated by Fenton reaction on the growth and cartilage differentiation in limb bud cell culture. AB - A consistent chondrogenesis takes place in micro high-density cultures of chick limb bud mesenchyme cells stage 22-24. The effect of an increased generation of OH(.) free radicals by Fenton reaction was tested in these cultures. Components of Fenton reaction (i.e., ferrous iron in form of ADP-Fe2+ complex in 0.1 mM concentration, or 0.2 mM H2O2, or the combination of these components with each other, as well as with 0.2 mM ascorbate) were supplemented to the culture medium after the first 24 h. ADP-Fe2+ complex resulted in a drastic decrease of the frequency and confluence of the cartilage nodules seen in light microscope, accompanied electron microscopically by a strikingly increased frequency of occurrence of lipofuscin-like, residual bodies in the cells. Biochemical methods revealed a significant decrease of both the DNA and glycosaminoglycan contents (to 48.6 and 20.7% of the controls, respectively), in day 6 cultures. H2O2 alone caused similar alterations of the cultures, whereas the combination of it with ADP-Fe2+ complex proved to be lethal for the cells. Ascorbate when added to the ADP-Fe2+-treated cultures displayed a slight protective effect for the glycosaminoglycan content but not for DNA. The results are interpreted in terms of free radical theory of aging. PMID- 3039156 TI - Intracisternal tubules in a case of chronic lymphocytic leukaemia. AB - In a case of chronic lymphocytic leukaemia we found that a few (less than 5%) of the circulating leukaemic lymphocytes contained inclusions of a type which do not appear to have been reported in the literature. The inclusions presented as solitary tubules or aggregates of parallel tubules lying within dilated cisternae of the rough endoplasmic reticulum and the perinuclear cistern. Some of the tubules were bounded by a single wall, others by a double wall. We also found a tubule bounded by three walls and another where a segment of the tubular profile was triple-walled and the remainder double-walled. We are inclined to think that the walls of these tubules are composed of membranes. The aetiogenic agent or agents responsible for the production of intracisternal tubules and their mode of formation are obscure. PMID- 3039157 TI - In vivo effectiveness of oral complexation agents in the management of iron poisoning. AB - The accidental ingestion of iron-containing preparations is relatively common in children, as are intentional overdoses with iron in adults. One of the most commonly used treatment modalities is the use of orally administered sodium bicarbonate or sodium hydrogen phosphate, which is thought to render iron less soluble and bioavailable by converting it to ferrous carbonate or ferrous phosphate. An in vivo parallel study utilizing 3 groups of 50 male Sprague-Dawley rats was designed to evaluate the effectiveness of oral complexation agents at five distinct time points. Each animal group was dosed with 300 mg/kg ferrous sulfate (60 mg/kg elemental iron), followed by a 5% solution of either sodium bicarbonate, sodium dihydrogen phosphate, or a control of distilled water. The animals were anesthetized in each group at separate time points of 0, 1, 2, 4, and 6 hours post iron load, and blood samples were taken to determine serum iron levels. There were no differences in serum iron levels between the control group and either complexing agent. This suggests that oral complexation in iron poisoning may be ineffective. PMID- 3039158 TI - Adrenocortical neoplasm in children. Ultrasound appearance. AB - The ultrasound appearances of three children presenting with virilization are described. All cases were also examined by computed tomography. The value of ultrasound in the initial detection of virilizing neoplasm and in follow-up studies for localization of tumor recurrence following initial surgery are outlined. The advantages of ultrasound include confirmation of extrarenal and extrahepatic location in longitudinal sections, and clear demonstration of invasion of hepatic veins, the inferior vena cava (IVC), and the right atrium on real-time scanning. Computed tomography is also useful, particularly in the localization of small lesions and the detection of pulmonary metastases. PMID- 3039159 TI - The four classes of endogenous murine leukemia virus: structural relationships and potential for recombination. AB - The process by which leukemogenic viruses are generated during the lifetime of certain strains of mice is poorly understood. We have therefore set out to define all the murine leukemia virus-related endogenous proviruses of HRS/J mice. We have cloned 34 different proviral fragments and their flanking cellular sequences. These have been characterized by restriction enzyme analysis, by fingerprinting in vitro-synthesized RNA, and by DNA sequencing. We conclude that all the proviruses can be assigned into one of four different classes: the previously characterized ecotropic, xenotropic, and polytropic viruses, as well as a new class we have termed modified polytropic viruses. The xenotropic, polytropic, and modified polytropic classes are closely related to one another, but as a group they differ considerably from the ecotropic class. Sequence analyses show that both polytropic and modified polytropic sequences can contribute env sequences to recombinant viruses. PMID- 3039160 TI - Genome segment reassortment between two serotypes of bluetongue virus in a natural host. AB - The occurrence of genome segment reassortment between two antigenically related orbiviruses was demonstrated in cattle. Individual virus clones were isolated by cell culture plaque assays directly from the blood of a calf infected with two serotypes of bluetongue virus. The majority (89%) of progeny viruses isolated from the calf represented reassortant viruses. A minimum of six genome segments participated in reassortment, with 16 unique reassortant constellations being identified. Such genome segment reassortment between unique, though antigenically related, orbiviruses has undoubtedly played a major role in generating the extensive phenotypic and genotypic diversity that is characteristic of this serogroup. PMID- 3039161 TI - Lack of competition results in efficient packaging of heterologous murine retroviral RNAs and reticuloendotheliosis virus encapsidation-minus RNAs by the reticuloendotheliosis virus helper cell line. AB - We constructed recombinant reticuloendotheliosis virus (Rev)-derived and murine leukemia virus-derived vectors to characterize the specificity of packaging retroviral RNAs in Rev proteins. Using this approach, we further localized the Rev encapsidation sequence (E) to a 144-nucleotide region and determined that there are sequences in both the 5' and 3' halves of this region which are necessary in cis for viral replication. We found that the Rev E, like the murine leukemia virus E (psi), is position independent (R. Mann and D. Baltimore, J. Virol. 54:401-407, 1986). Also, a 156-nucleotide region of the Rev intron enhanced replication in a cis-acting fashion in the presence, but not in the absence, of helper virus. Finally, we showed that packaging of E- and heterologous retroviral genomes occurred efficiently in the Rev helper cell in the absence of competing E-containing (E+) viral RNAs. PMID- 3039162 TI - Identification of the human papillomavirus type 6b L1 open reading frame protein in condylomas and corresponding antibodies in human sera. AB - Genital warts (condylomata acuminata) are among the most frequent sexually transmitted infections. Human papillomavirus type 6 (HPV-6), which is etiologically related to a majority of these lesions, has not been propagated in tissue culture. We generated two forms of HPV-6 viral antigens: a chemically synthesized oligopeptide (referred to as the C-terminal synthetic peptide) corresponding to residues 482 to 495 of the 500-amino-acid-long L1 open reading frame (ORF), and a bacterially expressed 54-kilodalton (kDa) fusion protein containing the N-terminal 13 amino acids encoded by the lambda bacteriophage cII gene followed by one vector-insert junctional residue and 462 amino acids of the L1 ORF sequence (residues 39 to 500). The cII-L1 fusion protein was specifically recognized by an antipeptide serum directed against the N-terminal 13 amino acids derived from the cII gene, an antiserum raised against the C-terminal synthetic peptide, and a genus-specific serum prepared by immunization with disrupted viral capsids. The 54-kDa fusion protein was purified, and the sequence of its first 36 amino acids was determined and found to be as predicted by the DNA sequence. Both the genus-specific anticapsid serum and the antiserum raised against the fusion protein identified authentic L1 ORF proteins in HPV-1-induced (58 kDa) and HPV 6/11-induced (56 kDa) papillomas. The synthetic peptide antiserum recognized the 56- to 58-kDa protein in HPV-6-induced warts, but not in HPV-1- or HPV-11 infected specimens. Using the fusion protein as antigen in immunoassays, we were able to detect the corresponding antibodies in human sera. PMID- 3039163 TI - The pseudorabies virus gII gene is closely related to the gB glycoprotein gene of herpes simplex virus. AB - We have looked for conserved DNA sequences between four herpes simplex virus type 1 (HSV-1) glycoprotein genes encoding gB, gC, gD, and gE and pseudorabies virus (PRV) DNA, HSV-1 DNA fragments representing these four glycoprotein-coding sequences were hybridized to restriction enzyme fragments of PRV DNA by the Southern blot procedure. Specific hybridization was observed only when HSV-1 gB DNA was used as probe. This region of hybridization was localized to a 5.2 kilobase (kb) region mapping at approximately 0.15 map units on the PRV genome. Northern blot (RNA blot) analysis, with a 1.2-kb probe derived from this segment, revealed a predominant hybridizing RNA species of approximately 3 kb in PRV infected PK15 cells. DNA sequence analysis of the region corresponding to this RNA revealed a single large open reading frame with significant nucleotide homology with the gB gene of HSV-1 KOS 321. In addition, the beginning of the sequenced PRV region also contained the end of an open reading frame with amino acid homology to HSV-1 ICP 18.5, a protein that may be involved in viral glycoprotein transport. This sequence partially overlaps the PRV gB homolog coding sequence. We have shown that the PRV gene with homology to HSV-1 gB encoded the gII glycoprotein gene by expressing a 765-base-pair segment of the PRV open reading frame in Escherichia coli as a protein fused to beta galactosidase. Antiserum, raised in rabbits, against this fusion protein immunoprecipitated a specific family of PRV glycoproteins of apparent molecular mass 110, 68, and 55 kilodaltons that have been identified as the gII family of glycoproteins. Analysis of the predicted amino acid sequence indicated that the PRV gII protein shares 50% amino acid homology with the aligned HSV-1 gB protein. All 10 cysteine residues located outside of the signal sequence, as well as 4 of 6 potential N-linked glycosylation sites, were conserved between the two proteins. The primary protein sequence for HSV-1 gB regions known to be involved in the rate of virus entry into the cells and cell-cell fusion, as well as regions known to be associated with monoclonal antibody resistance, were highly homologous with the PRV protein sequence. Furthermore, monospecific antibody made against PRV gII immunoprecipitated HSV-1 gB from infected cells. Taken together, these findings suggest significant conservation of structure and function between the two proteins and may indicate a common evolutionary history. PMID- 3039164 TI - Monoclonal antibody-aided characterization of cellular p220 in uninfected and poliovirus-infected HeLa cells: subcellular distribution and identification of conformers. AB - A monoclonal antibody directed against the Mr-220,000 subunit (p220) of the mRNA cap-binding complex has been prepared and used to analyze the sucrose gradient sedimentation and subcellular location of p220 and its poliovirus-induced cleavage products. The antibody reacted with p220 on immunoblots of cell lysates from uninfected cells, but only with several smaller polypeptides, the p220 cleavage products, in cell lysates from poliovirus-infected cells. The sedimentation of p220 antigens from uninfected or infected cells was analyzed by immunoblot and by enzyme-linked immunosorbent assay (ELISA) of sucrose gradient fractions. The results indicate that antibody reactivity was partially influenced by antigen conformation. Major forms of intact p220 and cleaved p220 were identified by immunoblot, and these had similar sedimentation properties. ELISA analysis of the same gradient fractions detected only uncleaved p220; p220 cleavage products were not recognized. Furthermore, the antibody recognized two forms of native uncleaved p220, one of which appeared to bind antibody with greater affinity. This result suggested the existence of conformational variants of p220. The differential reactivity of the antibody for cleaved versus uncleaved p220 served as a useful control during indirect immunofluorescence analysis to determine the subcellular distribution of p220 antigens. The distribution of p220 in uninfected cells was mainly cytoplasmic, but some nuclear antigens were also apparent. After poliovirus infection only the nuclear pattern remained. Disappearance of the cytoplasmic pattern confirmed the inability of the antibody to react with native p220 cleavage products. The cytoplasmic pattern also disappeared after human rhinovirus 14 infection, but not after mengovirus infection, results which correlated with the ability of human rhinovirus 14 and the inability of mengovirus to induce the cleavage of p220. The results demonstrate that p220 is not likely to be associated with the cytoskeleton and hint at the possibility of a partially nuclear location. PMID- 3039165 TI - Poliovirus proteinase 2A induces cleavage of eucaryotic initiation factor 4F polypeptide p220. AB - Poliovirus infection of HeLa cells induces rapid shutoff of host protein synthesis, whereas translation of poliovirus RNA is not inhibited. It is presumed that shutoff is the result of proteolytic cleavage of component p220 of eucaryotic initiation factor 4F. To study whether poliovirus proteinase 2A is involved in this cleavage, we translated synthetic RNAs that contained the coding region for poliovirus-specific polypeptides P1 and 2A in vitro and assayed for cleavage of p220. We report here that cleavage of p220 occurred in all cases when active proteinase 2A was translated and that disruption of the coding sequence of 2A by linker insertion or deletion prevented processing of p220 in vitro. Activity of 2A was determined by its ability to cleave at the P1-P2 site of a segment of the poliovirus polyprotein. We also constructed a plasmid in which the 3'-most 500 nucleotides of the nontranslated region of encephalomyocarditis virus were linked to the coding sequence for poliovirus polypeptide 2A. Translation of the RNA transcript of this clone was very efficient and yielded a fusion protein that included 2A; this polypeptide also induced cleavage of p220. In vitro translation in the presence of antibodies against 2A specifically inhibited processing of p220, whereas incubation of in vitro translation products with antibodies against 2A after translation was completed did not prevent proteolysis of p220. PMID- 3039166 TI - Insertion of adenovirus type 12 DNA in the vicinity of an intracisternal A particle genome in Syrian hamster tumor cells. AB - In the adenovirus type 12 (Ad12)-induced hamster tumor T1111(2) about 10 Ad12 genome equivalents were integrated at different sites. One of the integrated copies proved unstable and was lost from the cellular genome or rearranged upon passage of the cell line, H1111(2), established from this tumor. This unstable site of junction between the left terminus of Ad12 DNA and hamster DNA and the preinsertion site from BHK21 hamster cells was cloned, sequenced, and analyzed. The junction site showed several peculiarities. At the left terminus of Ad12 DNA, the first 64 nucleotides were deleted. At a distance of 127 nucleotides to the left from this junction site, an internal dispersed fragment of Ad12 DNA comprising nucleotides 1290 to 1361 of the authentic Ad12 DNA sequence was inserted into cellular DNA in an inverted orientation relative to the complete Ad12 genome that was located in its vicinity. The 127-nucleotide sequence between the intact Ad12 genome and the separate 72-base-pair (bp) Ad12 DNA fragment was cellular, but it was not identical to the preinsertion sequence at this location. The sequences flanking the termini of the dispersed 72-bp Ad12 DNA fragment were characterized by direct repeats of 9 or 10 nucleotides. To the left of Ad12 nucleotide 1361 in the separate 72-bp fragment, about 620 cellular nucleotides followed which were identical at the occupied and at the preinsertion sites. It was conceivable that the separate 72-bp Ad12 DNA fragment and the cellular sequence of 127 bp to its right had been transposed en bloc from another unknown location. Abutting the 620 nucleotides of cellular DNA to the left of this block, the 3'-terminal sequence of an endogenous, intracisternal A particle (IAP) genome of hamster cells was detected. The possible significance of the proximity of an IAP sequence to an inserted Ad12 genome with respect to the transformation event, to the instability at this site, or to the transcriptional activity of this region is not known. The 620 bp of cellular DNA between the 72-bp Ad12 DNA fragment and the end of the long terminal repeat of the hamster IAP sequence was apparently of a unique type. Transcriptional activity was not found in the approximate region between nucleotides -620 (to the left) and +350 (to the right) relative to the site of Ad12 DNA insertion, but was found outside these boundaries. PMID- 3039167 TI - Anti-idiotypic antibodies to a polyomavirus monoclonal antibody recognize cell surface components of mouse kidney cells and prevent polyomavirus infection. AB - Anti-idiotypic antibodies have been successfully used to identify and isolate the receptor for several cell ligands. To prepare an immunologic probe for identification of the polyomavirus receptor on mouse kidney cells, polyclonal antisera against antipolyomavirus antibodies were prepared in rabbits. Fab fragments of the previously characterized monoclonal antibody E7, which neutralizes polyomavirus infection, were used for immunization (S. J. Marriott and R. A. Consigli, J. Virol. 56:365-372, 1985). Sera containing the greatest anti-idiotype activity were identified by enzyme-linked immunosorbent assay (ELISA) and purified by a series of affinity columns. The anti-idiotypic antibodies recognized the E7 idiotope in an ELISA, and anti-idiotype binding could be inhibited by preincubation of E7 monoclonal antibody with polyomavirus virions. When mixed with anti-idiotype immunoglobulin G (IgG), E7 was no longer capable of binding or immunoprecipitating polyomavirus virions or neutralizing polyomavirus infection. Direct immunofluorescence showed anti-idiotype IgG reactivity with a cell surface component of mouse kidney cells. Anti-idiotype F(ab')2 effectively competed with polyomavirus for binding to mouse kidney cells and displayed binding kinetics similar to those of polyomavirus. Virus infection of mouse kidney cells was blocked in a dose-dependent manner following treatment of the cells with anti-idiotype IgG. The anti-idiotype identified several proteins (95, 50, and 24 to 31 kilodaltons) in an immunoblot of mouse kidney cell membrane proteins but reacted predominantly with a single 50-kilodalton protein in a radioimmunoassay. The anti-idiotype failed to react with virus proteins in three assays, including ELISA, immunoprecipitation, and immunoblotting. The implications of this work for future identification and characterization of the polyomavirus receptor on mouse kidney cells are discussed. PMID- 3039168 TI - Role of pseudorabies virus glycoprotein gI in virus release from infected cells. AB - The Bartha vaccine strain of pseudorabies virus has a deletion in the short unique (Us) region of its genome which includes the genes that code for glycoproteins gI and gp63 (E. Petrovskis, J. G. Timmins, T. M. Gierman, and L. E. Post, J. Virol. 60:1166-1169, 1986). Restoration of an intact Us to the Bartha strain enhances its ability to be released from infected rabbit kidney cells and increases the size of the plaques formed on these cells (T. Ben-Porat, J. M. DeMarchi, J. Pendrys, R. A. Veach, and A. S. Kaplan, J. Virol. 57:191-196, 1986). To determine which gene function plays a role in virus release from rabbit kidney cells, deletions were introduced into the genomes of both wild-type virus and the "rescued" Bartha strain (Bartha strain to which an intact Us had been restored) that abolish the expression of either the gI gene alone or both gI and gp63 genes. The effect of these deletions on the phenotype of the viruses was studied. Deletion mutants of wild-type virus defective in either gI or gI and gp63 behave like wild-type virus with respect to virus release and plaque size on rabbit kidney cells. Deletion of gI from the rescued Bartha strain, however, strongly affects virus release and causes a decrease in plaque size. We conclude that gI affects virus release but that at least one other viral function also affects this process. This function is defective in the Bartha strain but not in wild type virus; in its absence gI is essential to efficient release of the virus from rabbit kidney cells. PMID- 3039170 TI - Identification of proteins encoded by the L1 and L2 open reading frames of human papillomavirus 1a. AB - The human papillomavirus 1 (HPV-1) virion is composed of two virally encoded proteins: a 57,000-molecular-weight polypeptide (57K polypeptide), which is the product of the L1 open reading frame (ORF), and a 78K polypeptide, which is derived from the L2 ORF. The 57K (L1) product, which represents the major structural component, appears to be disulfide cross-linked in virus particles. The 78K (L2) protein is a minor component of the virion and does not appear to be disulfide linked either to the L1 gene product or to itself. Analysis of virus particles banding at different buoyant densities revealed differences in the L2 content of heavy-full and light-full virions. Antiserum prepared against a bacterially expressed fragment of the L1 ORF was found by immunofluorescence to cross-react with HPV-2 and bovine papillomavirus 1 virions in wart sections. No cross-reactivity was observed with antisera prepared against either the N- or C terminal halves of the L2-encoded protein. Similarly, antisera prepared against purified virus particles (disrupted and nondisrupted) reacted only with an expressed fragment of the L1 ORF and not with either L2-encoded polypeptides or proteins derived from the E1, E2, E4, E6, or E7 ORFs. This indicates that the L1 protein contains the papillomavirus common antigens. PMID- 3039169 TI - The membrane glycoprotein of Friend spleen focus-forming virus: evidence that the cell surface component is required for pathogenesis and that it binds to a receptor. AB - The leukemogenic membrane glycoprotein of Friend spleen focus-forming virus (SFFV) has an apparent Mr of 55,000 (gp55), is encoded by a recombinant env gene, and occurs on cell surfaces and in intracellular organelles. There is evidence that the amino-terminal region of gp55 forms a dualtropic-specific domain that is connected to the remainder of the glycoprotein by a proline-rich linker (C. Machida, R. Bestwick, B. Boswell, and D. Kabat, Virology 144:158-172, 1985). Using the colinear form of a cloned polycythemic strain of SFFV proviral DNA, we constructed seven in-phase env mutants by insertion of linkers and by a deletion. The mutagenized SFFVs were transfected into fibroblasts and were rescued by superinfection with a helper murine leukemia virus. Four of the mutants cause erythroblastosis. These include one with a 6-base-pair (bp) insert in the ecotropic-related sequence near the 3' end of the gene, two with a 12- or 18-bp insert in the region that encodes the proline-rich linker, and one with a 6-bp insert in the dualtropic-specific region. The other mutants (RI, Sm1, and Sm2) are nonpathogenic and contain lesions in dualtropic-specific region. The other mutants (RI, Sm1, and Sm2) are nonpathogenic and contain lesions in dualtropic specific sequences that are highly conserved among strains of SFFV. A pathogenic revertant (RI-rev) was isolated from one mouse that developed erythroblastosis 3 weeks after infection with RI. RI-rev contains a second-site env mutation that affects the same domain as the primary mutation does and that increases the size of the encoded glycoprotein. All pathogenic SFFVs encode glycoproteins that are expressed on cell surfaces, whereas the nonpathogenic glycoproteins are exclusively intracellular. The pathogenic SFFVs also specifically cause a weak interference to superinfection by dualtropic MuLVs. These results are compatible with the multidomain model for the structure of gp55 and suggest that processing of gp55 to plasma membranes is required for pathogenesis. The amino-terminal region of gp55 binds to dualtropic murine leukemia virus receptors, and this interaction is preserved in the SFFV mutants that cause erythroblastosis. PMID- 3039171 TI - Analysis of RNA synthesis of type 1 poliovirus by using an in vitro molecular genetic approach. AB - Membranous crude replication complexes (CRC) were isolated from poliovirus infected HeLa cells as recently described (N. Takeda, R.J. Kuhn, C.-F. Yang, T. Takegami, and E. Wimmer, J. Virol. 60:43-53, 1986). Viruses used to produce the CRC were poliovirus type 1 (Mahoney), [PV-1(M)], poliovirus type 1 (Sabin) [PV 1(S)], and four in vitro recombinants that were constructed from infectious cDNA clones. RNA synthesis in CRC was studied. No end-linked, full-length double stranded poliovirus RNA was detected in CRC regardless of whether nonionic detergent (Nonidet P-40) was added prior to incubation. Synthesis of VPg-pU and VPg-pUpU, two nucleotidyl proteins presumed to be involved in the initiation of RNA synthesis, was slower at 30 degrees C in CRC induced by PV-1(S) than by PV 1(M). This observation was used to design a pulse-chase experiment whose result suggested that synthesis of VPg-pUpU occurred by uridylylation of VPg-pU. Synthesis of VPg-pU(pU) was thermosensitive in CRC induced by PV-1(S). With CRC of recombinant viruses, the thermosensitive block covaried to nucleotide substitutions in PV-1(S) that mapped to the virus-induced RNA polymerase 3Dpol. We conclude that plus-stranded RNA synthesis in CRC does not proceed via hairpin structures. The results of VPg-pU----VPg-pUpU synthesis are consistent with a model in which VPg-pU is the primer of RNA synthesis mediated by 3Dpol. The data suggest that uridylylation of VPg or a precursor thereof may be catalyzed by 3Dpol itself, a mechanism resembling events occurring in adenovirus DNA replication. PMID- 3039172 TI - The role of Moloney murine leukemia virus RNase H activity in the formation of plus-strand primers. AB - On the basis of earlier studies with both detergent-disrupted virions (the endogenous reaction) and an in vitro reconstructed reaction, the RNase H activity associated with Moloney murine leukemia virus reverse transcriptase has been implicated in the generation of plus-strand RNA primers during reverse transcription. Here we used an oligonucleotide extension assay to show that the RNA primers remaining bound to the plus DNA strands initiated at the normal origin in the in vitro reaction are heterogeneous in length. This result indicates that, although a precise cleavage generates the 3' end of the priming RNA, RNase H exhibits less specificity at other break sites. During the endogenous reaction, a kinetic analysis of the synthesis of plus strands corresponding to different regions of the genome suggested that additional sites for the initiation of plus-strand DNA existed upstream of the normal origin. Direct analysis of fragments produced in the endogenous reaction, as well as in the in vitro reaction, confirmed the existence of upstream plus-strand initiation sites. Several of these sites were mapped to the nucleotide level by the oligonucleotide extension method. A comparison of the nucleotide sequences surrounding the upstream initiation sites with the sequence at the normal plus strand origin revealed common features, which suggests a mechanism for plus strand priming based on sequence recognition by the RNase H/reverse transcriptase protein. Although primer removal by RNase H is highly efficient for DNA fragments initiated at the normal origin, the RNA primers were inefficiently removed from the fragments initiated at the upstream sites. This result suggests that primer removal, like primer generation, involves sequence recognition by the enzyme. PMID- 3039173 TI - The role of envelope glycoprotein processing in murine leukemia virus infection. AB - The murine leukemia virus envelope protein is synthesized as a precursor molecule, Pr85env, which is proteolytically cleaved at an arginine residue to produce two mature envelope proteins, gp70 and p15(E). The results presented here indicate that mutation to lysine of the arginine found at the envelope precursor cleavage site results in a precursor which is cleaved with an efficiency at least 10-fold lower than the efficiency with which the wild-type protein is cleaved. This mutation has been used to investigate the requirement for envelope protein processing in various aspects of retroviral infection. Viruses produced by cells transfected with mutant proviral clones are approximately 10-fold less infectious than wild-type viruses. Mutant viruses are incapable of inducing XC cell syncytium formation and are 100-fold less efficient than wild-type viruses at rendering cells resistant to superinfection. Envelope glycoproteins bearing the lysine mutation are found in reduced amounts on the surface of infected cells, and as a result mutant virions contain significantly less envelope protein than do wild-type virions. The phenotypic effects of the processing mutation described here are most likely the result of this paucity of envelope glycoproteins in virions carrying the mutation. PMID- 3039174 TI - Simian virus 40-transformed human cells that express large T antigens defective for viral DNA replication. AB - Many types of human cells cultured in vitro are generally semipermissive for simian virus 40 (SV40) replication. Consequently, subpopulations of stably transformed human cells often carry free viral DNA, which is presumed to arise via spontaneous excision from an integrated DNA template. Stably transformed human cell lines that do not have detectable free DNA are therefore likely to harbor harbor mutant viral genomes incapable of excision and replication, or these cells may synthesize variant cellular proteins necessary for viral replication. We examined four such cell lines and conclude that for the three lines SV80, GM638, and GM639, the cells did indeed harbor spontaneous T-antigen mutants. For the SV80 line, marker rescue (determined by a plaque assay) and DNA sequence analysis of cloned DNA showed that a single point mutation converting serine 147 to asparagine was the cause of the mutation. Similarly, a point mutation converting leucine 457 to methionine for the GM638 mutant T allele was found. Moreover, the SV80 line maintained its permissivity for SV40 DNA replication but did not complement the SV40 tsA209 mutant at its nonpermissive temperature. The cloned SV80 T-antigen allele, though replication incompetent, maintained its ability to transform rodent cells at wild-type efficiencies. A compilation of spontaneously occurring SV40 mutations which cannot replicate but can transform shows that these mutations tend to cluster in two regions of the T antigen gene, one ascribed to the site-specific DNA-binding ability of the protein, and the other to the ATPase activity which is linked to its helicase activity. PMID- 3039175 TI - Assembly and processing of the disulfide-linked varicella-zoster virus glycoprotein gpII(140). AB - Varicella-zoster virus (VZV) specifies the synthesis of at least four families of glycoproteins, which have been designated gpI, gpII, gpIII, and gpIV. In this report we describe the assembly and processing of VZV gpII, a structural protein of an apparent Mr of 140,000, which is the homolog of gB of herpes simplex virus. For these studies, we used two anti-gpII monoclonal antibodies which exhibited both complement-independent neutralization activity and inhibition of virus induced cell-to-cell fusion. Pulse-chase labeling experiments identified a 124,000-Mr intermediate which was chased to the mature 140,000-Mr product when analyzed in nonreducing gels; in the presence of a reducing agent, the native gp140 was cleaved into two closely migrating species (gp66 and gp68). The biosynthesis of VZV gpII was further analyzed in the presence of the following inhibitors of glycoprotein processing: tunicamycin, monensin, castanospermine, swainsonine, and deoxymannojirimycin. All intermediate and mature forms were digested with endoglycosidases H and F, neuraminidase, and O-glycanase to further define high-mannose, complex, and O-linked glycans. Finally, the addition of sulfate residues was investigated. This characterization of VZV gpII revealed the following results. (i) gp128 and gp124 were early high-mannose forms, (ii) gp126 was an intermediate form with complex N-linked oligosaccharides, (iii) gp130 was a later intermediate with both N-linked and O-linked glycans, and (iv) the mature product gp140 contained a mixture of N-linked and O-linked glycans which were both sialated and sulfated. Further investigations indicated that gpII sulfation was inhibited by tunicamycin and castanospermine but not by deoxymannojirimycin or swainsonine. We also concluded that VZV gpII displayed many biological and biochemical properties similar to those of its herpes simplex virus homolog gB. PMID- 3039176 TI - Herpes simplex virus 1 protein kinase is encoded by open reading frame US3 which is not essential for virus growth in cell culture. AB - Earlier reports have described a novel protein kinase in cells infected with herpes simplex or pseudorabies viruses. These novel enzymes were characterized by their acceptance of protamine as a substrate and by their differential chromatographic behavior in anion-exchange chromatography. We report that this activity was not present in extracts of uninfected cells or of cells infected with a mutant constructed so as to contain a deletion in the US3 open reading frame mapping in the small component of herpes simplex virus 1 DNA. The activity was present in extracts of cells infected with wild-type virus and with a recombinant in which the US3 open reading frame had been rescued. Our results are consistent with the observation reported earlier that the coding sequences predict an amino acid motif common to protein kinases and lead to the conclusion that the US3 open reading frame encodes a virus-specific protein kinase that is not required for virus growth in cells in culture. PMID- 3039177 TI - Isolation and characterization of cDNA clones corresponding to transcripts from the BamHI H and F regions of the Epstein-Barr virus genome. AB - The Epstein-Barr virus (EBV) mutant P3HR1 is incapable of immortalizing B lymphocytes because of a 6.8-kilobase deletion in the BamHI W, Y, and H regions of the viral genome (M. Rabson, L. Gradoville, L. Heston, and G. Miller, J. Virol. 44:834-844, 1982). To characterize transcripts that are encoded in this region, poly(A)+ RNA from the EBV-transformed lymphoblastoid cell line JY was isolated, and this RNA was used to generate a cDNA library in lambda gt10. By screening 500,000 recombinant bacteriophages with the BamHI H fragment, we isolated 10 cDNA clones and characterized them in detail. One group of six cDNA clones was derived from a 2.9-kilobase early transcript encoded by the IR2 repeat element and showed restriction site polymorphism for the enzyme SmaI. The second group consisted of four cDNA clones, all of which contained the BamHI-H right reading frame (BHRF1), and used the polyadenylation signal at base pair 662 in the BamHI F fragment. Computer analysis of the hydrophobicity of the BHRF1 protein revealed that it is likely to be a membrane protein. Northern blotting experiments with RNA from an EBV producer line, B95-8, and a tightly latent lymphoblastoid B-cell line, IB4, revealed that BHRF1 is contained in at least two different mRNA species which can be detected during the latent cycle of EBV. These data and the recent characterization of a spliced transcript (containing five exons in common with other known latent messages [M. Bodescot and M. Perricaudet, Nucleic Acids Res. 14:7103-7113, 1986]) suggest that alternative splicing is used to generate transcripts containing BHRF1, as for the EBV nuclear antigen 1 transcripts. Furthermore, the observation that a potential oncogene activated in human follicular lymphomas is homologous to the BHRF1-encoded polypeptide (M. L. Cleary, S.D. Smith, and J. Sklar, Cell 47:19-28, 1986) suggests a possible role for this putative viral protein in EBV-induced growth transformation of B lymphocytes. PMID- 3039178 TI - A mouse model for poliovirus neurovirulence identifies mutations that attenuate the virus for humans. AB - A mutation in the genome of poliovirus type 3 that is known to reduce neurovirulence in humans similarly reduces neurovirulence in mice when incorporated into a mouse-adapted-human poliovirus recombinant. Viral recombinants with a uracil at nucleotide position 472 in the 5'-noncoding regions of their genomes are unable to replicate in the mouse brain. Viral recombinants with a cytosine at this position are neurovirulent in mice. Neurovirulence of poliovirus in mice may therefore prove to be a useful indicator of the genetic stability of new attenuating mutations created by site-directed mutagenesis. PMID- 3039179 TI - Levels of bovine papillomavirus RNA and protein expression correlate with variations in the tumorigenic phenotype of hamster cells. AB - Three independent cell lines were established from primary cultures of LSH hamster embryo cells infected with bovine papillomavirus type 1 (BPV-1). Although these cell lines differed in their in vitro saturation densities, none was capable of colony formation in soft agar. Interestingly, two cell lines (BPV-HE1 and BPV-HE3) were tumorigenic in nude mice, syngeneic hamsters, and allogeneic hamsters, whereas BPV-HE2 was not. All three cell lines contained similar numbers of the BPV-1 genome (approximately 50 to 200 copies per cell). However, the nontumorigenic BPV-HE2 cell line contained very low levels of BPV-specific RNA and only small amounts of the BPV-1 E5 transforming protein. The efficiency and rate of tumor formation by BPV-HE1 and BPV-HE3 correlated directly with the apparent amount of viral E5 protein. This analysis suggests that there is a threshold level of BPV protein synthesis required for tumorigenicity, there is a continuum of tumorigenic phenotypes which may depend upon the level of BPV protein expression, and BPV-transformed hamster cells can withstand allogeneic transplantation. PMID- 3039181 TI - Localization of calcium on the polyomavirus VP1 capsid protein. AB - Our laboratory has previously shown that the divalent cation Ca2+ is an integral part of the polyomavirus and plays a major role in stabilizing the intact virion structure. In this report, we show that calcium is sequestered on the major capsid protein VP1 of polyomavirus. The virion structural proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis before being transferred to nitrocellulose and probed with 45CaCl2. Autoradiography revealed 45Ca binding exclusively to VP1. Increasing the amount of VP1 transferred to the nitrocellulose resulted in a concomitant increase in 45Ca binding. 45Ca binding to VP1 could be reduced by competition with an excess of unlabeled CaCl2. Separation of the species of VP1 by two-dimensional gel electrophoresis before electroblotting and probing with 45CaCl2 revealed that all six species (A to F) bind the radiolabeled calcium. Formic acid cleavage of the 43-kilodalton (kDa) VP1 protein into 29-, 18-, and 16-kDa fragments before 45Ca-binding analysis revealed that only the 18- and 16-kDa carboxyl-terminal fragments of this protein bind 45Ca. PMID- 3039180 TI - Characterization of somatically acquired ecotropic and mink cell focus-forming viruses in lymphomas of AKXD recombinant inbred mice. AB - The DNA of lymphomas from 12 AKXD recombinant inbred mouse strains was analyzed to determine the presence of somatically acquired ecotropic and mink cell focus forming proviruses. Mink cell focus-forming proviruses were associated primarily with T-cell lymphomas, whereas ecotropic proviruses were associated with lymphomas of B-cell and myeloid lineages. A model based on the results is proposed to explain the variation in lymphoma types observed in different AKXD strains. PMID- 3039182 TI - Transcription of the cottontail rabbit papillomavirus early region and identification of two E6 polypeptides in COS-7 cells. AB - Cottontail rabbit papillomavirus (CRPV) early proteins are present at very low levels in virus-induced tumors and cannot be detected by immunological methods. Furthermore, cells in culture are not readily transformed by the virus. To overcome these difficulties in identifying and characterizing the putative transforming protein(s) coded by the E6 open reading frame, the early cottontail rabbit papillomavirus region was expressed under the control of the late simian virus 40 promoter. Mapping of the transcripts in transiently transfected COS-7 cells indicated that transcription was initiated in the late region of simian virus 40. Two E6-coded polypeptides were identified, representing translation products initiated at the first and second AUG codons. PMID- 3039184 TI - The extent and distribution of skeletal muscle necrosis after graded periods of complete ischemia. AB - The management of an acutely ischemic extremity requires knowledge of the response of skeletal muscle (the largest component of the lower limb) to prolonged periods of complete normothermic ischemia. We have used the canine gracilis muscle model to evaluate the extent and distribution of ischemic necrosis after 3 and 5 hours of ischemia and 48 hours of reperfusion. Each muscle was cut into six slices, and the extent and distribution of postischemic necrosis identified by means of nitroblue tetrazolium staining and 99mTc pyrophosphate uptake. After 3, 4, and 5 hours of ischemia the extent of necrosis was 2.0% +/- 0.9%, 30.3% +/- 6.0%, and 90.1% +/- 3.5% (mean +/- SEM), respectively. A statistically significant correlation exists between the extent of necrosis and the uptake of 99mTc pyrophosphate uptake per gram of tissue (y = 1574.9x - 8.4, r = 0.84, p less than 0.001). Most necrosis was centrally located and found in the thickest portion of the muscle. We conclude that there is a graded response in the extent of skeletal muscle necrosis related to the length of ischemic stress rather than an "all-or-none" phenomenon. This central distribution of necrosis makes the usual external evaluation of ischemic damage clinically unreliable. In addition, since there was no enveloping fascia in this model, a compartment release alone may not prevent the development of skeletal muscle necrosis. This knowledge of the response of skeletal muscle to ischemia may lead to an improved clinical approach to an extremity suffering a prolonged ischemic insult. PMID- 3039183 TI - Association of Epstein-Barr virus early antigen diffuse component and virus specified DNA polymerase activity. AB - The role of Epstein-Barr virus (EBV) early antigen diffuse component (EA-D) and its relationship with EBV DNA polymerase in EBV genome-carrying cells are unclear, EBV-specified DNA polymerase was purified in a sequential manner from Raji cells treated with phorbol-12,13-dibutyrate and n-butyrate by phosphocellulose, DEAE-cellulose, double-stranded DNA-cellulose, and blue Sepharose column chromatography. Four polypeptides with molecular masses of 110,000, 100,000, 55,000, and 49,000 daltons were found to be associated with EBV specified DNA polymerase activity. A monoclonal antibody which could neutralize the EBV DNA polymerase activity was prepared and found to recognize 55,000- and 49,000-dalton polypeptides. An EA-D monoclonal antibody, R3 (G. R. Pearson, V. Vorman, B. Chase, T. Sculley, M. Hummel, and E. Kieff, J. Virol. 47:183-201, 1983), was also able to recognize these same two polypeptides associated with EBV DNA polymerase activity. It was concluded that EBV EA-D polypeptides, as identified by R3 monoclonal antibody, are critical components of EBV DNA polymerase. PMID- 3039185 TI - Preventive strategies in sexually transmitted diseases for the primary care physician. US Preventive Services Task Force. AB - This study provides the background recommendations of the US Preventive Services Task Force for interventions by primary care physicians to prevent sexually transmitted diseases. Rationale for and data supporting use of barrier methods, epidemiologic treatment, contact tracing, patient education, prophylactic antibiotics, and disease reporting are discussed. Specific recommendations include those for gonorrhea, syphilis, human immunodeficiency virus infection, enteric infections, human papillomavirus infection, herpes simplex virus infection, and Chlamydia trachomatis infection. PMID- 3039186 TI - 'Chronic active Epstein-Barr virus infection'. PMID- 3039187 TI - Prognostic value and limitation of doughnut pattern of technetium-99m pyrophosphate myocardial uptake in acute anterior infarction. AB - The prognostic significance of the doughnut pattern of technetium-99m pyrophosphate myocardial uptake was evaluated in 140 patients with acute anterior infarction. There were significantly higher early complications, greater mortality and more severe hemodynamic abnormalities in the doughnut pattern group than in the non-doughnut pattern group. The former had a more depressed left ventricular ejection fraction and larger thallium-201 defect size (27.6 +/- 10.4 versus 40.0 +/- 13.5%, p less than 0.001 and 9.9 +/- 3.6 versus 5.6 +/- 3.3, p less than 0.001, respectively). There was, however, considerable overlap of the ranges of these variables for both groups. The patency rate of the infarct vessel during the acute phase of infarct in each group was similar (54.8 versus 45.2%). It is concluded that the prognostic value of the doughnut pattern may be limited to some extent by this overlapping and the presence of this pattern does not appear to correlate with the lack of residual blood flow to the infarcted area. PMID- 3039188 TI - Changes of Ca2+-ATPase and cytochrome oxidase activity of myocardial cell under early and late ischemia:--comparison with ultrastructural changes. AB - Using the histo- and cytochemical technique we assessed the Ca2+-transporting function of mitochondria (Mit) and sarcoplasmic reticulum (SR), and the ATP producing function of Mit in the ischemic myocardial cell of a dog's heart. In comparing ultrastructural ischemic changes, cytochrome oxidase (CO) and Ca2+ ATPase were cytochemically and histochemically measured when the myocardium was subjected to the ischemia of left anterior descending coronary artery occlusion for 15 min, and 60 min. After 15 min of occlusion the ischemic alterations consisted of a wide I band, decreased glycogen (G) and Mit swelling with a slight reduction of matrix density. Although CO activity was not reduced, Ca2+-ATPase had decreased mainly in Mit. Sixty min of ischemia resulted in loss of G, intermyofibrillar edema, marked Mit swelling with loss of matrix density and partial disruption of cristae, and dilatation of SR. Ca2+-ATPase activity was significantly reduced in Mit, SR and myofibrils. Although there was Mit swelling with partial disruption of cristae after 60 min of ischemia, CO activity was found to still exist in the remaining cristae. These findings suggest that intracellular organelle dysfunction progresses in the ischemic myocardial cell at different rates, and that disruption of intracellular Ca2+-homeostasis may occur early in the ischemic state. PMID- 3039189 TI - Roles of arachidonate metabolites in thrombus formation and leukocyte migration: analyses at microcirculatory level. AB - Thromboxane (TX) A2 is involved in platelet aggregation, which initiates thrombus formation. The platelet thrombi in microcirculation, however, are not homogeneous in nature. Two types of platelet thrombi, formed in an arteriole of the hamster cheek pouch, could be differentiated in sensitivity to indomethacin and PGI2. The "stable thrombus" was sensitive to indomethacin, and was not inhibited by PGI2, when applied after the formation, whereas the "ADP-induced thrombus" was sensitive to PGI2, but insensitive to indomethacin. These characteristics were reflected by two types of aggregation of composed platelets. Leukotriene B4 induced adhesion of the leukocytes on the venular wall in very low concentrations, when perfused over the microvasculature of hamster cheek pouch. The leukocyte adhesion on the venular wall was followed by migration into interstitial space. This adhesion was attributed to a change of leukocytes, rather than of the endothelial cell surface, as ascertained by selective microinjection of LTB4 with a glass capillary. LTB4 was also accumulated in the ischemic cardiac tissue after ligation of the rat coronary artery, followed by a peak in the accumulation of leukocytes. Inhibition of the LTB4 generation by a 5 lipoxygenase inhibitor not only suppressed the leukocyte count by 40%, but also the infarct size by 34.4%. PMID- 3039191 TI - [Effects of isoflurane anesthesia and surgery on the endocrine function in man]. PMID- 3039190 TI - Anti-thrombotic and anti-atherogenic action of eicosapentaenoic acid. AB - Effects of dietary supplementation with highly purified EPA (1.8-2.7 g/day) for 16 weeks on platelet and red blood cell function and serum lipids concentration were investigated in patients with various thrombotic diseases. Decreases in platelet aggregation, thromboxane formation in platelets, platelet retention and whole blood viscosity, increased red blood cell deformation and prolongation of bleeding time were observed in the present study. In addition a reduction in serum cholesterol and triglyceride concentrations was noted in patients with hyperlipidemia after EPA ingestion. Some clinical improvements such as improvement of diabetic gangrene or peripheral vascular occlusive disease were observed. These results indicate that dietary supplementation of purified EPA may be beneficial for prevention and treatment of cerebro- and cardiovascular diseases. PMID- 3039193 TI - [A case of breast cancer with a chondrosarcomatous appearance]. AB - A case of a breast tumor which showed the appearance of invasive ductal carcinoma and chondrosarcoma is reported. The patient was a 36-year-old female. Simple mastectomy and lymph node dissection were performed for the tumor. Local recurrence and metastases to the contralateral breast, lymph nodes, subcutaneous tissue, and bones arose after the operation. She died of the disease 2 years and 3 months after the operation. It is thought that this tumor had formed as a result of metaplasia of the carcinomatous tissue, since a histological transition between the carcinoma and sarcoma was demonstrated. PMID- 3039192 TI - [A case of small cell carcinoma of the esophagus]. AB - A small cell carcinoma of the esophagus is a rare entity; only 101 cases have been reported in the world literature. In a 59-year-old man, complaining of dysphagia, a tumorous lesion was detected in the esophagus at the upper GI series. The histologic diagnosis of biopsy revealed an undifferentiated carcinoma or a malignant lymphoma. Preoperative radiotherapy was carried out, effectively decreasing the large tumorous lesion in size. Thereafter an esophagectomy and an esophagogastric anastomosis were done. The histologic diagnosis of the resected specimen revealed a small cell carcinoma of an intermediate cell type. The tumor cells failed to show cytoplasmic argyrophilia and an ultrastructural examination did not display any cells containing membrane-bound neurosecretory granules. PMID- 3039194 TI - [Colorectal cancer and synchronous adenomatous or malignant polyps--factors influencing its incidence]. AB - A retrospective study was made of 433 patients with colorectal cancer who were operated on between January, 1971 and September, 1986. Two-hundred and forty eight synchronous polyps (226 adenomas and 22 carcinomas) were found in 128 patients (29.6%). The incidence of polyps varied according to the patient's age, sex, family history of colorectal cancer, and also according to the macroscopic classification, depth of infiltration, and the number of the primary tumors. Of these, macroscopic classification seemed to be the most important factor determined by multivariate analysis. The incidence of synchronous polyps of the protuberant type were the highest (50%), as compared to the ulcerating type (28%) and the infiltrating type (19%) (p less than 0.03). PMID- 3039195 TI - [A case of primary signet ring cell carcinoma of the breast]. AB - A rare case of primary signet ring cell carcinoma of the breast is reported. The patient was a 54-year-old unmarried female (G 0, P 0, AB 0) with a several months' history of inflammatory reactions, such as infection, erosion, and a swelling of her entire left breast. She died of this disease in 6 months despite an operation and combined chemotherapy (CMF). Her serum CEA level had well reflected the course of her treatment. Histopathologically, a primary lesion and axillary lymph nodes were found to be filled with typical signet ring cells, whose cytoplasm showed positive staining for PAS and Alcian blue stains as well as CEA. This case, both clinically and histopathologically, displayed the peculiar characteristics of signet ring cell carcinoma, which differs from other types of breast cancer. PMID- 3039196 TI - [An autopsy case of mucin-producing pancreatic cancer]. AB - A 85-year-old woman with a mucin-producing pancreatic cancer is reported. The patient, who had been diabetic for 2 years, was admitted in March of 1984 to our hospital because of a fever and cough. On admission, a hard tumor was found on the right hypochondric region. Ultrasonogram and computed tomography revealed a cystic pancreas head tumor containing mucin and a dilated pancreatic duct. During an ERCP examination, Vater's papilla was found to be enlarged and a biopsy showed papillary adenocarcinoma. Also, the levels of CEA and CA 19-9 were elevated 11.3 ng/ml and 1300 U/ml, respectively. On Feb. 13, 1985, she died due to panperitonitis that resulted from a perforated duodenal ulcer. Microscopic examination of the pancreas showed a papillary adenocarcinoma producing mucin, a dilated pancreatic duct and atrophy of the islets. No metastatic lesion was found. The pathogenesis and the clinical characteristics of the mucin-producing pancreatic cancer are discussed. PMID- 3039197 TI - [A case of perineal (extramammary) Paget's disease]. AB - A case of perineal Paget's disease in a 67-year-old male patient is reported. The perineal polyp, having shown no increase in size for 3 years, wks resected. The microscopic features of the polyp was comprised of clusters of a pale, large tumor cell (Paget cell) only in the epidermis. The skin that was resected around the polyp showed almost the same histological appearance as the polyp, except for an infiltration of tumor cells forming a small gland structure on the minimal region of the dermis. Both the Paget's cells and these tumor cells showed positive for mucin staining. Ultrastructurally, we confirmed two types of Paget's cells, one containing much granules and the other lacking in these granules but rich in other organelles. Further, the Paget's cells formed lumen and intercellular canaliculus, suggesting a differentiation to the eccrine sweat gland. From our findings and a review of the literature, we favor presuming that extramammary Paget's disease without a cancer of the neighbouring organs originates from the epidermis and extends to the dermis. PMID- 3039198 TI - [Photoradiation therapy (PRT). 7. Clinical study of photoradiation therapy: clinical application and efficacy. d. Brain neoplasms]. PMID- 3039199 TI - Phase 2 study of high dose etoposide (VP16-213) in hepatocellular carcinoma. AB - A phase II study of etoposide (VP16-213) in hepatocellular carcinoma was conducted among 18 Chinese patients. There was no observable tumour response but the treatment was relatively well tolerated with an overall median survival interval of 74 days. PMID- 3039200 TI - Detection of gastrin-releasing peptide mRNA in small cell lung carcinomas and medullary thyroid carcinomas using synthetic oligodeoxyribonucleotide probes. AB - Human gastrin-releasing peptide (GRP) mRNA was detected in the tumor tissues of medullary thyroid carcinomas and small cell lung carcinomas using synthetic oligodeoxyribonucleotides as hybridization probes. The amount of GRP mRNA was estimated by radiodensitometric hybridization assay. A good correlation was found between the amount of GRP mRNA and the concentration of immunoreactive GRP in the tumor tissues. PMID- 3039201 TI - [Electrodiagnosis and SEP study of peripheral nerve, root and plexus lesions]. PMID- 3039202 TI - [Mechanism of regulation of blood coagulation]. PMID- 3039203 TI - [Antigen detection with monoclonal antibodies: 2. Detection of human rotavirus subgroups]. PMID- 3039204 TI - [Magnetic resonance imaging of hepatic tumors using a rapid pulse sequence method]. PMID- 3039205 TI - [A case of hepatocellular carcinoma with adrenal metastasis]. PMID- 3039206 TI - [Myelopathy due to intercostal artery embolization with lipiodol: a case report]. PMID- 3039207 TI - [Diagnostic imaging of head and neck tumors. Tumors arising from and adjacent to the temporal bone]. PMID- 3039208 TI - [Radiology of head and neck tumors in children]. PMID- 3039209 TI - [A long-taper catheter device]. PMID- 3039210 TI - [Epidermoid cyst of the sole and human papillomavirus]. PMID- 3039211 TI - [A case of hepatocellular carcinoma which developed from type B hepatitis and presented marked hypoglycemia]. PMID- 3039212 TI - [A study of gamma-interferon production induced by interleukin 2 in peripheral blood mononuclear cells of patients with hepatocellular carcinoma]. PMID- 3039213 TI - [Quantitative evaluation of left-sided valvular regurgitation by gated single photon emission computed tomography]. PMID- 3039214 TI - [Monitoring of LV function in coronary care unit--evaluation of the effect of a vasodilator by the nuclear cardiac probe]. PMID- 3039215 TI - Inhibition of arsenazo III Ca transient with deuterium oxide in frog twitch fibers at a resting sarcomere length. AB - Arsenazo III calcium transients during twitches of frog skeletal muscle fibers were recorded in H2O Ringer (control) and deuterium oxide Ringer (test). In ten fibers, the amplitude of the test at a sarcomere length of 2.5 microns was reduced to 57% of the control and accompanied by extremely diminished tension responses. The latency was also increased to 158%. PMID- 3039216 TI - Potassium secretion in the guinea pig distal colon. AB - Active K+ secretion in guinea pig distal colon was studied in vitro in Ussing chambers. Changes in short-circuit current (Isc), transepithelial conductance (Gt), and the unidirectional flux of 42K from serosa to mucosa (JKsm) were determined under a variety of physiological and pharmacological conditions. Mucosal tetraethylammonium (TEA) (10 or 30 mM) increased Isc, while it decreased Gt and JKsm. Seroal bumetanide (10(-4) M) also caused an increase in Isc and decreases in Gt and JKsm. The effects produced by TEA were abolished in the presence of bumetanide. The increase in Isc caused by bumetanide was greatly reduced by mucosal high-K+ conditions. Isoproterenol (10(-6) M), a beta adrenergic agonist, elicited a decrease in Isc and an increase in Gt. Responses to isoproterenol (10(-6) M) were reduced or abolished by mucosal TEA, serosal bumetanide, serosal ouabain (10(-4) M), and serosal Na+- or Cl(-)-free conditions. Mucosal high K+ also reduced the isoproterenol-induced decrease in Isc. These results suggest that K+ secretion is electrogenic and dependent on the bumetanide-sensitive cotransport in the basolateral membrane. K+ exit across the apical membrane might be conductive and sensitive to TEA. In addition, beta adrenergic agonists might stimulate K+ secretion. PMID- 3039217 TI - Down-regulation of central serotonin S2 receptors after repeated treatment with quinupramine in rats. AB - The present studies were undertaken to determine whether the repeated administration of quinupramine caused down- or up-regulation of beta-, alpha 2 adrenergic, serotonin S2, imipramine and muscarinic cholinergic receptors, as had been demonstrated for tricyclic and atypical antidepressant drugs. Quinupramine administered at 10 mg/kg (p.o.) twice daily for 10 days caused a down-regulation of serotonin S2 receptors in the frontal cortex of the rat as determined by [3H]ketanserin binding. However, quinupramine did not alter the binding populations of beta-adrenergic, muscarinic cholinergic and alpha 2-adrenergic receptors in the rat brain as determined by the Scatchard analysis of the [3H]ligand binding data. Differing from quinupramine, imipramine caused down regulation of beta-adrenergic and serotonin S2 receptor bindings, and it caused slight but significant up-regulation of muscarinic cholinergic receptor bindings. These results show that the antidepressant activity of quinupramine is associated with the central serotonin system, but not with the beta-adrenergic system. Accordingly, quinupramine, chemically one of the typical tricyclic antidepressant drugs, seems to be pharmacologically one of the atypical antidepressant drugs, and it was suggested that the central serotonin system plays an important role in the antidepressant activity of quinupramine. PMID- 3039218 TI - Hypotensive effect of SA446, an angiotensin converting enzyme inhibitor, in 2 kidney, 1-clip renal hypertensive and normotensive dogs. AB - Hypotensive effects of SA446, an angiotensin converting enzyme (ACE) inhibitor, and effects on the renin-angiotensin system were evaluated in conscious normotensive and 2-kidney, 1-clip renal hypertensive dogs. SA446 (1 mg/kg, p.o.) remarkably inhibited the pressor response to angiotensin (Ang) I between 1 and 6 hr after the administration in normotensive dogs. SA446 significantly decreased blood pressure at 10 mg/kg, p.o., in normotensive dogs. During repeated administration of SA446 (100 mg/kg/day, p.o.) for 13 weeks, the blood pressure was lowered, and the pressor response to Ang I and plasma ACE activity were strongly inhibited. ACE activities in the aorta and kidney were also inhibited. Plasma renin activity and plasma Ang I concentration increased by repeated SA446 application, while plasma aldosterone concentration decreased. The hypotensive effect of SA446 (5 mg/kg, p.o.) was more potent in 2-kidney, 1-clip renal hypertensive dogs than in normotensive dogs. SA446 had longer inhibitory effects on the pressor response to Ang I and more potent hypotensive effects than captopril. The hypotension caused by SA446 appears to be associated mainly with an inhibition of ACE in plasma and also in the vascular wall. PMID- 3039219 TI - [Tracheobronchoplastic procedures in the treatment of lung cancer]. PMID- 3039220 TI - [Evoked electrospinogram of the bulbocavernosum reflex recorded from epidural space]. PMID- 3039221 TI - [Study of the continence mechanism--the role of the beta adrenergic receptor in the proximal urethra]. PMID- 3039222 TI - [A case of aniridia-Wilms' tumor syndrome]. PMID- 3039223 TI - Pathological characteristics of salmonid herpesviruses and antibody production of the virus-infected fish. PMID- 3039224 TI - Quantitative single radial immunodiffusion of foot-and-mouth disease viral antigens using convalescent cattle sera. PMID- 3039225 TI - Cross-reactivity among infectious bronchitis viruses in enzyme-linked immunosorbent assay. PMID- 3039226 TI - Fatal herpesvirus infection in a litter of puppies. PMID- 3039227 TI - Micro-neutralization test with canine coronavirus for detection of coronavirus antibodies in dogs and cats. PMID- 3039228 TI - [Does guar gum really provide a plus in the treatment of type II diabetes? (Myth or reality)]. PMID- 3039229 TI - [Role of ion movements and changes in membrane potential in beta cells for the control of insulin secretion]. PMID- 3039230 TI - [Metabolism of lipoproteins: role of specific receptors]. PMID- 3039231 TI - [Medical topics: Ockelbo disease/myoglobin and myocardial infarction]. PMID- 3039232 TI - Platelet activating factor (PAF) as a mediator of injury in nephrotoxic nephritis. AB - Release of acetyl glyceryl ether phosphorylcholine, platelet-activating factor (PAF), has been demonstrated to be associated with glomerular inflammatory damage in acute serum sickness. Moreover, PAF can increase glomerular permeability to proteins and induce mesangial contraction. Thus PAF might be a good candidate as a mediator of glomerular damage. However the in vivo evidence that PAF might cause glomerular injury is lacking. To evaluate if PAF has a major role in promoting glomerular inflammatory reaction and fibrin deposition, we studied the effect of a molecule, L-652,731, which blocks the PAF receptor, on the evolution of an experimental model of anti-glomerular basement membrane (anti-GBM) glomerulonephritis (GN). GN was initiated by sheep-anti-rabbit nephrotoxic serum. A proliferative GN regularly occurred in which heavy proteinuria, intra and extracapillary proliferation of resident and inflammatory cells and fibrin deposition in Bowman's capsule were the prominent findings. Our results showed that the PAF receptor antagonist reduces the glomerular damage in anti-GBM GN, supporting the hypothesis that PAF is indeed a mediator of glomerular inflammatory reaction. PAF receptor blocking prevented renal function deterioration in the early phase of the disease, probably preserving glomerular hemodynamics. In the delayed phase of the disease the PAF receptor antagonist reduced proteinuria and prevented renal function deterioration and fibrin deposition. These effects appear to be mediated by an inhibitory action of PAF receptor blocking on macrophage accumulation and activation. PMID- 3039233 TI - Renal target structures in acute allograft rejection: a histochemical study. AB - With an aim to investigate the relative sensitivity of various renal structures to allograft rejection, we analyzed the histochemical reaction intensity of seven enzymes prominently displayed in various rat kidney components, and correlated the expression of these enzymes both to the degree of intra-graft inflammation and to the expression of class II MHC antigens in graft capillary endothelial cells. Syngeneic transplants and normal renal tissue were used as controls. At the peak of inflammation, on the fifth day after transplantation, adenosine triphosphatase activity of vascular endothelial cells was strongly reduced in the peritubular capillary endothelium of the allograft, moderately in the glomerular endothelium but very little in the endothelium of arteries and veins. Lactate dehydrogenase, succinate dehydrogenase, isocitrate dehydrogenase, alkaline phosphatase, acid phosphatase and glucose-6-phosphatase activities were moderately reduced in the proximal tubular cells of the allograft and even less in the distal tubular cells. The results suggest that the prime target of the host immune attack is the intertubular capillary endothelium, whereas the distal tubular cells are relatively insensitive to immune injury. PMID- 3039234 TI - Dietary NaCl determines severity of potassium depletion-induced metabolic alkalosis. AB - It is uncertain whether, in humans, potassium depletion can cause or sustain metabolic alkalosis of clinically important degree in the absence of coexisting known alkalosis-producing conditions. Previously we found, in normal humans ingesting abundant NaCl, that dietary K+ depletion alone can induce and sustain a small decrease in blood acidity and increase in plasma bicarbonate concentration; we hypothesized that more severe alkalosis was prevented by mitigating mechanisms initiated by renal retention of dietary NaCl that was induced by K+ depletion. To ascertain the acid-base response to dietary K+ depletion under conditions in which the availability of NaCl for retention is greatly limited, in the present study of six normal men we restricted dietary K+ as in the previous study except that intake of NaCl was maintained low (2 to 7 mEq/day, Low NaCl Group) instead of high (126 mEq/day, High NaCl Group). Plasma acid-base composition and renal net-acid excretion (NAE) did not differ significantly between groups during the control period. In the steady state of K+ depletion (days 11 to 15 of K+ restriction), neither plasma K+ concentration (2.9 +/- 0.9 mEq/liter vs. 3.0 +/- 0.1 mEq/liter) nor cumulative K+ deficit (399 +/- 59 mEq vs. 466 +/- 48 mEq) differed significantly between groups. During K+ restriction, persisting metabolic alkalosis developed in both groups, which was more severe in the Low NaCl Group: increment in [HCO3-]p, 7.5 +/- 1.0 mEq/liter versus 2.0 +/- 0.3 mEq/liter, P less than 0.001; decrement in [H+]p, 5.5 +/- 0.6 nEq/liter versus 2.9 +/- 0.4 nEq/liter, P less than 0.003. A significantly more severe alkalosis in the Low NaCl Group was evident at all degrees of K+ deficiency achieved during the course of the 15 days of K+ restriction, and the severity of alkalosis in the Low NaCl Group correlated with the degree of K+ deficiency. During the generation of alkalosis (days 1 to 7 of K+ restriction), NAE increased in the Low NaCl Group whereas it decreased in the High NaCl Group. During the maintenance of alkalosis (days 11 to 15), NAE stabilized in both groups after it returned to values approximating the control values. In both groups, urine Cl- excretion decreased during K+ restriction even though Cl- intake had not been changed, with the result that body Cl- content increased negligibly in the Low NaCl Group (28 +/- 6 mEq) and substantially in the High NaCl Group (355 +/- 64 mEq).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3039235 TI - Immunohistochemical expression of malignant fibrous histiocytoma induced by 4 (hydroxyamino) quinoline 1-oxide in Wistar rats. PMID- 3039236 TI - The proteins associated with human T-cell leukemia virus type-I (HTLV-I) and their functions. PMID- 3039237 TI - [Surgical treatment of small cell cancer of the lung]. PMID- 3039238 TI - [Limited resection in cancer of the lung]. PMID- 3039239 TI - [Cases of tumors of the liver]. PMID- 3039240 TI - [Insuloma of the pancreas in a diabetic]. PMID- 3039241 TI - [Endoscopic laser photodestruction of tumors as a method of palliative treatment of lung cancer]. PMID- 3039242 TI - [Surgical treatment of glomus tumors of the hand and forearm]. PMID- 3039243 TI - [Barre-Mason tumors]. PMID- 3039244 TI - [Successful treatment of a Wilms' tumor]. PMID- 3039245 TI - [Immunotherapy of posttraumatic vitreous hemorrhage]. PMID- 3039246 TI - Corticotropin releasing hormone in plasma of patients with Cushing's disease. PMID- 3039247 TI - [Hormonal response of the bodies of cosmonauts after completion of brief space flights]. AB - After short-term (7-day) space flights the following parameters were measured in blood of cosmonauts: cortisol, ACTH, aldosterone, thyrotropic hormone, thyroxine, triiodothyronine, somatotropic hormone, insulin, testosterone, cyclic nucleotides, prostaglandins, activities of the kallikrein-kinin, fibrinolytic and coagulatory systems, and plasma renin activity; in addition, renal excretion of aldosterone and total 17-oxycorticosteroids was determined. It was demonstrated that after the short-term flights the acute period of adaptation was accompanied by a moderate activation of the renin-angiotensin-aldosterone system, adrenal glucocorticoid function, pancreatic insular apparatus, kallikrein-kinin system as well as increased cyclic AMP which is suggestive of a moderately expressed stress reaction. PMID- 3039248 TI - [Features and mechanisms of the effect of adrenaline and noradrenaline on the pumping function of the heart during hypokinesia]. AB - The effect of epinephrine (E) and norepinephrine (NE) on cardiac output (CO) and heart rate (HR) of hypokinetic and control rats was studied. It was found that in the hypokinetic animals the E effect on CO was significantly weaker than in the controls while the NE effect was stronger. In most experiments the response to NE in the hypokinetic animals was greater than that to E. The mean HR increase was not significantly different in the two groups but in the hypokinetic rats the response to E was less expressed. These data give reason to believe that the lower E effect on CO in the hypokinetic animals is associated with diminished alpha-adrenergic effects on the heart. PMID- 3039249 TI - A dot-immunobinding assay on nitrocellulose with psoralen inactivated Herpesvirus simiae (B virus). AB - An enzyme-immunoassay performed with Herpesvirus simiae (B virus) and H. simplex antigens inactivated with a psoralen derivative and long-wavelength ultraviolet light irradiation is described. Although B virus is a known human pathogen requiring extreme care in its handling, the use of inactivated antigens in the test allows its performance without biosafety containment. The test utilizes nitrocellulose sheets dotted with antigen for the assay of antibody against B virus in nonhuman primate sera. Antigen-antibody complexes are detected visually as red dots by the use of enzyme-conjugated antiglobulin second antibody and a substrate that produces an insoluble product. The test is more rapid, sensitive and specific than the serum neutralization test it is intended to replace. Of 150 macaque monkey sera tested, 83 were negative by the enzyme and neutralization tests, 56 were positive by both tests and 11 were positive by enzyme-assay but negative by neutralization. Positive sera reacted with both simian and human viral antigens in the enzyme assay but with greater intensity against B virus. Absorption with H. simplex removes reactivity with this virus without reducing the B virus response. PMID- 3039250 TI - Prevalence of feline leukemia virus infection in random source laboratory cats. AB - Three years of quarantine data involving the prevalence of Feline Leukemia Virus (FeLV) in laboratory cats (Felis catus) was evaluated. Testing was performed using a commercially available ELISA system. Annual prevalence of infection ranged from 5.4 to 10.7% of the 937 cats tested. Results obtained with the enzyme linked immunosorbent assay (ELISA) system compared well with tests performed using an immunofluorescent antibody assay system (IFA). Seasonal periodicity was noted with higher infection rates among animals received in April. Holding period at the supplier and gender did not influence disease prevalence. The availability of a simple test system, the unacceptability of FeLV infected cats for research, the danger of transmission and a higher prevalence of infection than earlier reports indicated that supplier and user evaluation of research cats for FeLV would be prudent. PMID- 3039251 TI - Silica hazard of caisson construction in Hong Kong. PMID- 3039252 TI - Conjugated forms of [3H] alpha, 25-dihydroxyvitamin D3 in rat bile. AB - When small doses of [3H]D3, [3H]25-OHD3 and [3H]alpha, 25-diOHD3 were administered intravenously to rats 6.3 +/- 1.1% (means +/- SEM, n = 4), 9.7 +/- 0.9% (n = 6) and 12.8 +/- 2.6% (n = 8), respectively, of the administered radioactivity was excreted in bile. The radioactive biliary conjugated metabolites were analysed by ion exchange chromatography: in the case of all 3 substrates about 30% of metabolites were found to be cationic on the basis of their being retained on sulphopropyl-Sephadex G-25 (H+-form) when applied in 70% methanol. The balance of the metabolites were neutral and anionic and were analysed on TEAP-Lipidex: in the case of 1 alpha, 25-diOHD3 the following metabolite classes were detected (on the basis of co-elution with authentic standards) (in order of quantitative importance): taurine conjugates, neutral metabolites, monosulphates, glucuronides, carboxylic acids, glycine conjugates and disulphates. Alkaline hydrolysis of the taurine and glycine conjugates yielded products 60% of which now chromatographed as carboxylic acids. Hydrolysis of the glucuronide and monosulphate fractions indicated significant levels of mixed conjugation yielding some products which now chromatographed as glycine and taurine conjugates, respectively. The nature of the cationic conjugates was not elucidated but they had the following properties: they could be hydrolysed by alkali to yield non-cationic radioactive metabolites (these released metabolites were heterogeneous as judged by TEAP-lipidex chromatography); they were partially hydrolysed to non-cationic forms by beta-glucuronidase; and on reverse-phase HPLC they had an elution profile that was significantly different to that of histidyl , ornithyl- or lysyl-calcitroic acid. PMID- 3039253 TI - Prognosis after resection of non-small cell lung cancer of the right middle lobe. AB - In a previous study at Roswell Park Memorial Institute, it was noted that in patients whose resected non-small-cell lung cancer had originated in the right middle lobe, the prognosis was much worse than when it had originated in any other lobe. A review of a longer period and exclusion of patients who might introduce a bias resulted in a group of 18 patients whose fate was compared to the findings reported in other comparable series. Three out of those are alive at 8-96 months after operation. Five-year survival was 22% (30% counting only survivors of the operations). These findings indicate that the results of surgery for non-small-cell carcinoma of the right middle lobe fall within the lower range of lung cancer generally. PMID- 3039254 TI - Primary liver cancer in a referral hospital in Hawaii. AB - This is a retrospective review of 43 patients who had primary liver cancer diagnosed during 1974-1983. Patients' ages ranged from 27 to 84 years (median 52.5). Nine of 39 patients with hepatoma were females, while two of the four patients with cholangiocarcinoma were women. Hepatitis surface antigen was positive in 90% tested, and 62% had cirrhosis. Also, 60-65% were heavy users of alcohol and cigarettes. Alpha-fetoprotein was elevated in one of four white patients, and in six of eight patients of other races (75%). Tissue diagnosis was obtained by peritoneoscopy in 16, by percutaneous biopsy in 7, by laparotomy in 9, and at autopsy in 11. Only one of 11 patients who were explored has his lesion resected. About half of the cases diagnosed antemortem died 1 month or less after diagnosis. The median survival of hepatoma patients who had no specific treatment or systemic chemotherapy was 2 months. Two patients who received chemotherapy in conjunction with occlusion of the hepatic artery lived 16 to 19 months. PMID- 3039255 TI - A glucagon-secreting pancreatic alpha islet cell tumor presenting as spinal cord compression. AB - We describe a patient with a pancreatic islet carcinoma presenting with spinal cord compression owing to vertebral metastases. Subsequent studies demonstrated a typical islet cell carcinoma by light microscopy. By electron microscopy, the neurosecretory granules were morphologically suggestive of glucagon production. Radioimmunoassay studies revealed markedly elevated levels of serum glucagon. Notably, the patient did not exhibit the characteristic glucagonoma syndrome. This case exemplifies clearly that elevated levels of immunoreactive neuropeptide hormones are not necessarily associated with overt hormonal syndromes. Possible mechanisms for explaining this apparent discrepancy include the production of immunoreactive molecules with weak or absent systemic biological activity. Nevertheless, the determination of immunoreactive hormone levels in neuroendocrine neoplasms is an extremely effective adjunct method for their diagnosis and monitoring. PMID- 3039256 TI - The absolute density of neurotransmitter receptors in the brain. Example for dopamine receptors. AB - Since the absolute density of dopamine receptors can vary in disease, it is essential to establish the normal values for the absolute densities of D1 and D2 receptors in the brain. Absolute densities are most conveniently reported in units of picomoles per gram of original tissue, readily permitting their comparison to data obtained by positron emission tomography in patients. The density of D1 receptors is approximately 120 pmol/g in the rat striatum and 19 pmol/g in the human striatum. The density of D2 receptors is about 32 pmol/g in the rat striatum and 14 pmol/g in the human striatum, these values being determined by Teflon-glass homogenization and the centrifugation method. The customary Polytron-homogenization procedure results in a loss of about 9% of the D2 receptors in rat tissue and about 28% in human tissues; filtration results in a further loss of about 12%. There is general agreement between the in vitro and in vivo densities, but only if the receptors are measured by the amount of radioisotope specifically displaced. PMID- 3039257 TI - Ultrastructural observation and glucose-6-phosphatase determination on the repair of rat experimental hepatic lesions by qing kai ling No. 1. PMID- 3039258 TI - Phenotypic, cytogenetic and molecular characterization of a new B-chronic lymphocytic leukaemia (B-CLL) cell line. AB - A lymphoid cell line was established from a patient with B-cell chronic lymphocytic leukaemia (B-CLL) by infecting blood lymphocytes with Epstein-Barr virus (EBV). Immunoglobulin gene rearrangement studies and the presence of a chromosome marker (isochromosome 17q) provided the formal proof that the line has originated from the neoplastic B cells. The morphology and phenotype indicate that the EBV-induced cell line has reached a plasma cell-like stage of differentiation. PMID- 3039259 TI - Quantitative enzyme determination; a parameter for leukemic cell differentiation. AB - In acute myeloid leukemia the leukemic cells are thought to be blocked in their normal differentiation. The stage of differentiation is the basis for the FAB classification. Induction of cell differentiation is a new and promising development in the treatment of some myeloproliferative diseases. The criteria for cell maturation and differentiation are almost all based on the morphology of the leukemic cells. In this study we tried to specify the maturation stage of the leukemic cells by quantitative enzyme analysis. In the HL60 (promyelocytic) cell line cells we determined the following enzymes: myeloperoxidase; alpha naphthyl acetate esterase; alpha naphthyl butyrate esterase and lactate dehydrogenase. The enzyme profiles obtained after culturing the cells in the presence of the differentiation inducing agents 1:25 dihydroxy vitamin D3 and DMSO were compared with several cytochemical and functional parameters. The results obtained in this study show that quantitative enzyme analysis is a useful tool in the study of myeloid or monocytic differentiation. PMID- 3039260 TI - [Apropos of a case of neurologic bilharziasis]. PMID- 3039261 TI - [Malignant tumors of the testis in 1986]. PMID- 3039262 TI - [Type 1 hypocalcemic pseudo-deficiency rickets caused by renal 25 hydroxycholecalciferol 1-hydroxylase deficiency (apropos of 1 case)]. PMID- 3039263 TI - Age related differences in immunocompetence and incidence of mammary adenocarcinoma in murine mammary tumor virus-infected C3H/Bi mice. AB - In breeder C3H/Bi female mice, infected neonatally by murine mammary tumor virus (MTV), the incidence of spontaneous mammary tumors is greater than 95% between 5 and 9 months of age. In young (2-3 months) female the probability for developing a tumor in the next month is negligible, higher than 80% in mice of middle age (5 6 months) but lower than 4% in aged (10-12 months) females. The age-related changes of some immune functions of spleen cells from these tumor free female mice have been evaluated. While the proliferative capacity of cells to Phytohemagglutinin (PHA) increases, the T cell-dependent antibody response against sheep red blood cells (SRBC) and the antibody-dependent cellular cytotoxicity (ADCC) are significantly decreased in 5-6-month-old mice as compared to the young (2-3 months) female mice. The antibody response against SRBC and the mitogenic response to PHA decline markedly in 10-12-month-old mice but the ADCC increases in this group of mice. In addition, assays with monoclonal anti-Lyt-1 and anti-Lyt-2 antibodies indicate that percentage of Lyt 1- 2+ cells (suppressor and cytotoxic T cells) is lower in 10-12-month-old female as compared to 5-6 month-old animals. These results show that the immune alterations observed in 10 12-month-old C3H/Bi mice are not closely associated with an increase in incidence of spontaneous tumors and suggest that a high non-T killer cell activity could protect some of these older C3H/Bi female mice against mammary tumor development. PMID- 3039264 TI - Acylphosphatase levels of human erythrocytes during cell ageing. AB - Human erythrocytes contain an acylphosphatase isoenzyme, whose concentration during cell ageing was determined by an enzyme immunoassay. Erythrocytes were age fractionated by isopycnic centrifugation in "Percoll" density gradients. Acylphosphatase concentration was found to rise with the increase of cell density. Maximum values were attained in the mature erythrocytes and there was only a slight decrease in the older cells. Acylphosphatase activity in human erythrocytes of different ages followed a similar pattern, a finding which was confirmed for rabbit red cells with different levels of reticulocytes. The most probable mechanism for the increase of acylphosphatase content and activity during red cell maturation appears to be a de novo synthesis of this enzyme during the reticulocyte stage and its storage, virtually unaffected, in the mature erythrocytes. The possible consequences of increased acylphosphatase levels on the metabolic modifications associated with erythrocyte ageing are discussed. PMID- 3039265 TI - Biology of human papillomavirus (HPV) infections and their role in squamous cell carcinogenesis. AB - Current data implicating the role of human papillomavirus (HPV) infections in squamous cell carcinogenesis may be summarised as follows: animal models have shown that PVs can induce malignant transformation; HPV involvement in both benign and malignant human squamous cell tumours has been demonstrated by morphological, immunohistochemical and DNA hybridisation techniques; HPV infections in the genital tract are venereally transmitted and are associated with the same risk factors as cervical carcinoma; the natural history of cervical HPV lesions is similar to that of CIN, namely, they have the potential to develop into carcinoma in situ; malignant transformation of PV-induced lesions seems to depend on virus type and the physical state of its DNA, e.g., whether or not it is integrated in the host cell DNA; malignant transformation most probably requires synergistic actions between the PVs and chemical or physical carcinogens, or other infectious agents; genetic disposition (at least in animals) significantly contributes to malignant transformation; immunological defence mechanisms of the host are probably capable of modifying the course of PV infections (efficacy in man remains to be elucidated). Many details of the molecular mechanisms, however, still remain to be clarified. Although BPV1 is capable of transforming fibroblasts, the way that papillomaviruses transform epithelial cells is unclear. Which gene is capable of inducing the limited cell proliferation in benign lesions? No model systems exist to provide the answer nor to elucidate the progression to malignant cells and then to invasive cancer. Improved tissue culture systems for in vitro differentiation of keratinocytes should help in elucidating the biology of papillomaviruses and their interaction with cell division and differentiation. PMID- 3039266 TI - Simple and sensitive CO2 laser visual screen. AB - The present paper describes a simple and sensitive CO2 laser visual screen which is made from silica gel with cobalt chloride as indicator. The screen can directly and almost immediately display a clear mode pattern of an exposing CO2 laser beam, it can respond to laser intensities ranging from 80 mW/cm2 to 26 W/cm2. It is suitable for the observation of both high-power lasers, which are used for surgery, and very low power ones, which are used for biostimulation. PMID- 3039267 TI - [Rational diagnosis and therapy of irritable colon]. AB - The irritable bowel syndrome is a common motility disorder of the gut characterized by constipation, diarrhea and abdominal pain. Symptoms are markedly influenced by psychological factors. The diagnosis is based on typical symptoms and exclusion of organic diseases. Psychological support by the physician is an important part of the patients' treatment. High fiber diets and bulking agents may be prescribed in addition. Antispasmodic and antidiarrheal drugs should be given only the shortest time possible, while psychotropic drugs are seldom necessary. PMID- 3039268 TI - Field detection of early neuritis in leprosy. PMID- 3039269 TI - Heterogeneity of alpha 1 receptors associated with vascular smooth muscle: evidence from functional and ligand binding studies. AB - The nature of the alpha 1 receptor associated with rabbit aorta has been examined in functional and receptor binding studies. In isolated aortic rings the dose response curve for (-)metaraminol was not parallel to that of (-)epinephrine, ( )norepinephrine or (-)phenylephrine. Following inactivation of a portion of the alpha receptors with phenoxybenzamine, the occupancy versus response relationship for metaraminol, in contrast to the other test agonists, was biphasic. These results suggest the possibility that metaraminol interacts with different functional groups on the alpha 1 receptor than the other test agonists. In microsomes prepared from frozen aorta, metaraminol bound to two classes of sites (KH = 0.41 +/- 0.12 microM, KL = 39.1 +/- 7.1 microM) labelled by the selective alpha 1 antagonist [3H] prazosin. Similar binding characteristics were observed in microsomes prepared from aorta shipped in serum on ice or aorta from animals killed in our laboratory. Norepinephrine also bound to two sites on the alpha receptor in all three preparations tested (KH = 0.06 +/- 0.01 microM, KL = 5.09 +/- 2.4 microM; estimates from frozen aorta). The Scatchard plot of [3H]prazosin binding to microsomes prepared from frozen aorta was curvilinear. Estimates of the affinities and site densities were 49.6 +/- 15.3 pM and 44.8 +/- 11.8 pmol/gm protein and 1.0 +/- 0.2 and 43.8 +/- 17.4 pmol/gm for the high and low affinity sites, respectively. These data are consistent with the idea that there are subtypes of the alpha 1 receptor. PMID- 3039270 TI - Differences of binding characteristics of non-selective opiates towards mu and delta receptor types. AB - [3H]ET (etorphine), which is considered either as an "universal" ligand or a mu agonist, interacts with identical affinities KD = 0.33-0.38 nM to hybrid cells and rabbit cerebellum, pure delta and mu-enriched opioid receptor preparations, respectively. In rat brain tissue, [3H]ET binding is inhibited by DAGO (Tyr-D-Ala Gly-(Me)-Phe-Gly-ol), a mu selective agonist, in a competitive manner without apparent modification of the maximal number of sites. Furthermore, even at a DAGO concentration (300 nM) which should be sufficient to block [3H]ET interaction with mu sites, no variation in the total capacity of the tritiated ligand is observed. In contrast, DTLET (Tyr-D-Thr-Gly-Phe-Leu-Thr), a delta-preferential agonist, blocks [3H]ET binding in rat brain at a concentration able to saturate delta-sites. At higher concentrations, where DTLET cross reacts with mu-sites, this ligand exhibits similar properties to those of DAGO. These data are very different from those obtained with [3H]EKC (ethylketocyclazocine), another "universal" ligand, the binding properties of which are easily explained by the occurrence in rat brain tissue of independent sites exhibiting pharmacological profiles of mu, delta and kappa sites. Our results underline the possible misinterpretation of binding data obtained by using [3H] etorphine as a non selective ligand. PMID- 3039271 TI - Kinetic alterations of cytochrome c oxidase in carbon tetrachloride induced cirrhotic rat liver. AB - In a study of the chronic effects of CCl4 on the respiratory activities of rat liver mitochondria, the content of cytochrome c oxidase increased from 0.077 +/- 0.010 (nmol/mg protein) for normal rats to 0.101 +/- 0.009, and its specific activity increased from Vmax = 345 +/- 24 (e-/s/cytochrome aa3) to 431 +/- 19 in mitochondria of CCl4 treated rats. There was a slight increase in Km for cytochrome c from 5.63 +/- 0.08 microM to 7.79 +/- 0.80. These results would strongly suggest that an appreciable decrease in the steady state concentration of ATP in hepatic cells of CCl4 treated rats brought about a compensatory increase in the overall activity of cytochrome c oxidase. However, when the rate of oxygen uptake by mitochondria was measured in the presence of rotenone and tetramethyl-p-phenylene-diamine with NADH as substrate, the specific activity in CCl4 treated rats was lower than that of normal rats (Vmax = 345 +/- 31 (e /s/cytochrome aa3), as compared to Vmax = 408 +/- 21) in spite of the increased activity of cytochrome c oxidase. This phenomenon was ascribed to a decrease in the activity of NADH cytochrome b5 reductase in the mitochondrial outer membrane due to CCl4 treatment. PMID- 3039272 TI - Role of peripheral and central opioid activity in analgesia induced by restraint stress. AB - Rats subjected to prolonged restraint showed an increase in tail flick latency which outlasted the period of restraint by 15 min. This restraint could be blocked but not reversed by 1 mg/kg of naltrexone hydrochloride given subcutaneously. Naltrexone methobromide, administered subcutaneously in doses of 10 or 25 mg/kg, did not block the analgesia indicating that peripheral opioid receptors were probably not involved. Naltrexone hydrochloride was shown to have no effect on brain tryptophan uptake in restrained rats, a neurochemical event which had previously been shown to be critical to restraint-induced analgesia. PMID- 3039273 TI - The regulation of GABAergic receptors by a novel family of endogenous neuropeptides. PMID- 3039274 TI - Opioid peptides in man--analytical aspects. AB - The opioid peptides present extraordinary problems for analysis, since they are structurally homologous yet substantially different. Within each of the three systems, proopiomelanocortin, proenkephalin and prodynorphin, a whole set of peptides of successively smaller sizes are generated. The biological relevance of the different peptides is not fully understood. Analytical strategies may either be specifically directed to individual peptide species or to identification of total system output. PMID- 3039275 TI - Synthesis, purification and biological evaluation of porcine corticotropin releasing factor. AB - The 41-residue sequences of recently identified porcine corticotropin-releasing factors [Ile40]pCRF and [Asn40]pCRF were assembled on a benzhydrylamine resin support. Deprotection and cleavage from the resin were accomplished by HF treatment. The crude peptides were purified by HPLC. The homogeneity of the final materials, obtained in 0.2% and 0.4% overall yield for [Ile40]pCRF and [Asn40]pCRF respectively, was assessed after the isolation by HPLC and amino acid analysis. Both sequences of the synthetic 41-residue pCRF stimulated the release of corticotropin (ACTH) from superfused rat pituitary cells on a column, the responses being related to a log-dose of CRF in the range of 1-20 ng/ml. [Ile40]pCRF and [Asn40]pCRF also augmented the in vivo release of ACTH in rats pretreated with chlorpromazine, morphine and Nembutal. [Ile40]pCRF appeared to be equipotent to ovine CRF and about twice as active as [Asn40]pCRF. The data indicate that synthetic porcine [Ile40]pCRF and [Asn40]pCRF have high biological activity. PMID- 3039276 TI - Inhibitory action of spermidine on formyl-methionyl-leucyl-phenylalanine stimulated inositol phosphate production in human neutrophils. AB - The effect of the polyamine, spermidine, on formyl-methionyl-leucyl-phenylalanine stimulated hydrolysis of polyphosphoinositides was examined in purified human neutrophils by measurement of inositol phosphate production from radioactively labelled inositol. Spermidine inhibited formyl-methionyl-leucyl-phenylalanine stimulated inositol phosphate production by neutrophil in a dose dependent manner. Inhibition of formyl-methionyl-leucyl-phenylalanine stimulated inositol phosphate accumulation by spermidine was maximal at 10 microM and the IC50 value for this effect was 4.2 microM spermidine. This action of spermidine, thought to be mediated by a membrane component other than phospholipase C, may reflect a control mechanism modulating the response of the polyphosphoinositide system. PMID- 3039277 TI - Catecholaminergic control of plasma growth hormone and thyrotropin levels in hypophysectomized rats bearing ectopic pituitary transplants. AB - Transplantation of the anterior pituitary to an ectopic site leads to stimulation of PRL secretion and suppression of the release of other adenohypophyseal hormones. We have previously reported that precursors and blockers of catecholamine synthesis can affect PRL release from the ectopic pituitary. In the present study we have measured the effects of L-3,4-dihydroxyphenylalanine (DOPA), DL-threo-dihydroxyphenylserine (DOPS), alpha-methyl-para-tyrosine (alpha mpt) and diethyldithiocarbamate (ddc) on plasma growth hormone (GH) and thyrotropin (TSH) levels in hypophysectomized rats with pituitary transplants under the renal capsule. In these animals, peripheral plasma GH levels were elevated by a precursor (DOPA) and reduced by a blocker (alpha-mpt) of catecholamine synthesis. Plasma TSH levels were increased by a precursor (DOPS) and reduced by a blocker (ddc) of norepinephrine synthesis. We suspect that GH and TSH present in the circulation of pituitary-grafted animals were derived, in part, from the ectopic pituitary tissue and suggest that the small but detectable secretion of hormones other than PRL in this animal model is under the control of endogenous catecholamines. PMID- 3039278 TI - Opioid action of the intestine: the importance of the intestinal mucosa. AB - Drug effects on the intestine are traditionally explained in terms of action on the muscle layers and the nerves that control them. This is particularly true in the case of the opioids but research starting two decades ago has identified the intestinal mucosa as the site of action of the antidiarrhoeal opioids. Continued research using the intestinal mucosa offers a fresh approach to solving some old problems. For example it could lead to more confident predictions to be made about the wanted and unwanted effects of opioid drugs on the intestine and may help to find better drug treatments for alleviating withdrawal diarrhoea in addicts. Eventually it may help to explain how the general process of opioid dependence occurs at a cellular level. PMID- 3039279 TI - Heterogeneity of muscarinic receptors coupled to phosphoinositide breakdown in guinea pig brain and peripheral tissues. AB - Muscarinic receptors coupled to phosphoinositide hydrolysis (PI) are present in guinea pig bladder and colon. Compared to rat cerebral cortex, an extensively studied muscarinic/PI turnover system, all agonists were more potent and efficacious in both bladder and colon. The "M1-selective antagonists", pirenzepine and dicyclomine, were much more potent (Ki = 1-5 nM) and selective (300 to 500-fold) at both rat and guinea pig brain and guinea pig colon receptors, compared to PI-coupled receptors in guinea pig bladder. In contrast, "M2-selective antagonists", AF-DX 116 and HHSiD, were 2-6 fold more potent in bladder than in brain, while HHSiD was very potent in the colon (50 times more potent than in brain). These results suggest a pharmacological heterogeneity of PI-linked muscarinic receptors. If muscarinic receptors with a low affinity for pirenzepine are defined as M2, these results show that the guinea pig bladder contains PI-linked M2 muscarinic receptors, whereas the guinea pig colon contains PI-linked M1 receptors. PMID- 3039280 TI - Structural derivatives of pindolol: relationship between in vivo and in vitro potencies for their interaction with central beta-adrenergic receptors. AB - Although (-)-125I-iodopindolol (IPIN) can be used to label beta-adrenergic receptors in the central nervous system (CNS) in vivo, use of this ligand for receptor imaging studies in humans may be limited due to its relatively poor penetration into the CNS. A series of derivatives related to pindolol was therefore studied in an effort to determine the factors that might influence the penetration and interaction of these compounds with central beta-adrenergic receptors in vivo. Evaluation of the ability of these derivatives to displace the binding of IPIN in the brain upon systemic administration provides an assessment of whether the derivatives penetrate and interact with central beta-adrenergic receptors in vivo. Multiple regression analyses showed that the most important factor which influences the ability of the pindolol derivatives to penetrate into the brain and interact with beta-adrenergic receptors in vivo is the affinity of the derivatives for binding to beta-adrenergic receptors in vitro. Both lipophilicity and the molecular weights of the derivatives are important secondary factors which influence their in vivo potency. PMID- 3039281 TI - Distribution of calcium activated neutral proteinase (mM CANP) in myelin and cytosolic fractions in bovine brain white matter. AB - The activity of calcium-activated neutral proteinase (mM CANP) was determined in homogenate, myelin and supernatant of bovine brain corpus callosum. The enzyme activity in homogenate and myelin was increased eleven and thirteen-fold respectively by Triton X-100. Myelin prepared by the method of Norton and Poduslo as well as by a modified method, was shown to contain most (more than 50%) of homogenate mM CANP activity. The specific activity was highest in myelin, and increased almost three times more than the homogenate. Supernatant only contained 17% of enzyme activity. It is concluded from these studies that mM CANP is tightly bound to the membrane and predominantly associated with the myelin sheath. PMID- 3039282 TI - Calcium-dependent proteolysis in the myocardium of rats subjected to stress. AB - Activity of a calcium-dependent neutral protease (calpain II) and its specific endogenous inhibitor was investigated in the myocardium of rats subjected to different stressors: cold, anaesthesia, 24 and 48 h starvation and food restriction for 7 and 14 days. Enzyme and inhibitor activities were determined in the 37,200 g supernatant of homogenates prepared from the free left ventricular wall of the heart. The specific activity of the myocardial calcium-dependent proteinase increased in all rats exposed to stressful stimuli, reaching maximum values in animals starved for 48 hours. Decrease in the specific activity of the inhibitor accompanied the changes in enzyme activity. Differences from normal control values were statistically significant in the starved animals and in animals fed a restricted diet for 7 or 14 days. These observations suggest that interaction between calpain II and its specific inhibitor plays a role in the regulation of the enzyme activity and furthermore, that stressful stimuli lead to increased calcium-dependent proteolysis in the myocardium. PMID- 3039283 TI - Hepatocellular carcinoma associated with second primary malignancy. AB - In a consecutive series of 562 patients with pathologically documented hepatocellular carcinoma (HCC), 12 patients (2.1%) were found to have a second primary malignancy elsewhere. Of these, eight were male, four female, with ages ranging from 49 to 76 years (mean 60.7 +/- 9.3 years). The associated cancers included eight cases of gastric cancers, an esophageal cancer, a stump cancer, a case of myelocytic leukemia and a histiocytic lymphoma. Eight cases had both malignancies diagnosed at the same admission. In the other four, HCC were detected after an interval of 4, 10, 12 and 39 months. Two had received previous chemotherapy or radiotherapy. Fifty percent of the 12 patients were hepatitis B surface antigen (HBsAg) positive. Their liver function tests and alpha fetoprotein levels were consistent with those of patients with HCC alone. The results suggest that the occurrence of HCC concomitant with or arising after another primary malignancy in Taiwan is not rare. PMID- 3039284 TI - Immunohistochemical localization of ras p21 and carcinoembryonic antigens (CEA) in cholangiocarcinoma. AB - An expression of ras p21 proteins on cholangiocarcinoma (CC) (intrahepatic bile duct carcinoma) cells was examined by an immunoperoxidase method using an appropriate dilution of mouse monoclonal antibody RAP-5, with which no positive staining was obtained in livers with normal histology. Of 44 CCs examined 39 were positive for the antigens; well-differentiated adenocarcinoma usually showed a diffuse weak, cytoplasmic staining in nearly all tumor cells with the same staining intensity, while in moderately and poorly differentiated adenocarcinoma the expression of p21 varied markedly in intensity from cell to cell in the same cell nest. The number of positive cells decreased with the grade of tumor, and no or little staining was observed in undifferentiated areas. These findings indicate that the expression of ras p21 antigens was lost with increasing dedifferentiation of tumor cells. Carcinoembryonic antigens (CEA) were positive in 42 of 44 CCS. Well-differentiated adenocarcinoma expressed CEA along the apical surfaces of the tumor glands. With the dedifferentiation of tumor cells, the expression of CEA became prominent not only at the apical surfaces but also on the basolateral surfaces and in the cytoplasms, and further in the surrounding stromal tissue. There was no clear-cut correlation between the expression of p21 antigens and the production of CEA in CCs. PMID- 3039285 TI - [Dose multifractionation in radiotherapy of lung cancer of different histological types]. AB - A study was made of immediate and short-term results of multifractionated radiotherapy of 120 patients with inoperable lung cancer. On discontinuation of radiotherapy the frequency of complete and pronounced tumor regression was significantly higher in radiosensitive small-cell lung carcinoma whereas during radiation exposure the correlation between a degree of tumor regression and tumor histological structure was absent as a result of an increase in the number of primary local cure in patients with squamous-cell and glandular carcinoma. A study of short-term therapeutic results confirmed the efficacy of the use of multifractionated irradiation in all main histological types of lung cancer. PMID- 3039286 TI - [The state of immunity and the cyclic nucleotide system in the radiotherapy of breast cancer]. AB - Indices of the T-system of immunity were lowered in breast cancer patients. Small fractional radiotherapy resulted in its greater suppression. A tumor revealed low pathomorphosis accompanied by a sharp increase in the levels of cyclic nucleotides. Large fractional radiotherapy caused no changes in the state of the immune system correlating with high radiation pathomorphosis in a tumor and a decrease in a cGMP level. PMID- 3039287 TI - [Giant lymph node hyperplasia (Castleman's disease) with peripheral nerve diseases and syndrome of inappropriate secretion of antidiuretic hormone]. PMID- 3039288 TI - Expression of human tissue-type plasminogen activator from lytic viral vectors and in established cell lines. AB - We have used two kinds of vectors to express a cDNA of human tissue plasminogen activator (t-PA) in mammalian cells. In one case, cDNAs inserted into vectors based on bovine papilloma virus were introduced into cultured murine cells and cell lines were established that efficiently and continuously secrete enzymatically active t-PA into the medium. Second, the t-PA gene was used to replace the sequences of the simian virus (SV40) genome that code for the viral coat proteins. Virus stocks were generated and used to infect a stable line of cultured simian cells. During the resulting lytic infection, expression of the t PA gene is governed by the potent SV40 late promoter and enzymatically active t PA accumulates rapidly in the medium. We have used these two vector systems to analyze the biosynthesis and transport of recombinant t-PA and to compare its properties with those of "natural" t-PA secreted by the Bowes line of human melanoma cells. t-PA secreted from all three sources is identical in specific activity (approximately 20,000 units/mg) despite differences in patterns of terminal glycosylation. Furthermore, non-glycosylated t-PA synthesized in the presence of tunicamycin was secreted efficiently and was indistinguishable in specific activity from glycosylated t-PAs. PMID- 3039289 TI - Growth factor receptors and cell transformation. PMID- 3039290 TI - A comparison of simian rotavirus SA11 preparations maintained in different laboratories. AB - Preparations of simian rotavirus SA11 maintained in different laboratories were compared with each other by polyacrylamide gel electrophoresis of genomic RNA. Differences in the migration of genome segments 4, 5 and 7 allowed the classification of eight virus preparations into four electrophoretic types. PMID- 3039291 TI - Size variation in group A streptococcal M protein is generated by homologous recombination between intragenic repeats. AB - M protein, a major surface protein and virulence factor for the group A streptococcus, exhibits extraordinary size variation in strains of the same serotype (Fischetti et al. 1985). RNA sequence analysis of spontaneous M protein size variants shows that deletion mutations arise in a single strain by homologous recombination events between intragenic tandem repeats. Similar deletion and duplication events also occur in serial streptococcal isolates from a single patient and among related strains in a recent outbreak. We discuss how homologous recombination events can lead to the generation of antigenic variation. PMID- 3039292 TI - Construction of a series of ompC-ompF chimeric genes by in vivo homologous recombination in Escherichia coli and characterization of their translational products. AB - OmpC and OmpF are major outer membrane proteins and although they are homologous proteins, they function differently in several respects. As an approach to elucidate the submolecular structures that determine their differences, we have constructed a series of ompC-ompF chimeric genes by in vivo homologous recombination between these two genes, which are adjacent on a plasmid. The recombination sites in the chimeric genes were localized by means of restriction endonuclease analysis and nucleotide sequence determination. Most of the chimeric gene products were accumulated in the outer membrane. One of the chimeric gene products, with a fusion site in a central region between the OmpC and OmpF proteins, was normally expressed but not accumulated in the outer membrane. The trimeric structures of some of the chimeric gene products appeared to be extremely unstable in a SDS solution. From these results, domains contributing to the formation of specific structures in which the OmpC and OmpF proteins differ were identified. Bacterial cells possessing the chimeric gene products were also investigated as to their sensitivity to phages that require either OmpC or OmpF as a receptor component. With the aid of the chimeric gene products, the immunogenic determinants for three anti-OmpC monoclonal antibodies were found to be localized at different portions of the OmpC polypeptide: the N-terminal, central and C-terminal portions, respectively. PMID- 3039293 TI - Deletion analysis of the mannopine synthase gene promoter in sunflower crown gall tumors and Agrobacterium tumefaciens. AB - We have used deletion mutagenesis to analyze a TR-DNA promoter from the octopine type Ti plasmid pTiB6806. The promoter for the gene encoding mannopine synthase (mas) was cloned upstream of the bacterial kanamycin-resistance gene neomycin phosphotransferase II (NPT II). Bal31 deletion mutagenesis was used to generate deletion derivatives of the mas/NPTII gene beginning 1353 bp upstream of the initiation of transcription and extending to 120 bp downstream from the mRNA start site. Deletions that left intact 318 bp upstream of transcription initiation had no detectable effect on the ability of tumors harboring the deletion to synthesize correctly initiated mRNA or to grow on the kanamycin analogue G418. Deletion to-138 destroyed the ability of sunflower crown gall tumors to grow on G418 although low levels of the mas/NPTII transcript were detected in one tumor line. Deletions that left only 57 bp upstream of transcription initiation allowed neither growth on G418 nor detectable mas/NPTII synthesis, even though the CCAAT and TATAA homologies were intact. The mas promoter is functional in Agrobacterium tumefaciens and we present data concerning the effects of the Bal31 deletions on the growth of A. tumefaciens on kanamycin. PMID- 3039294 TI - Nucleotide sequence of the pyrimidine specific carbamoyl phosphate synthetase, a part of the yeast multifunctional protein encoded by the URA2 gene. AB - Yeast URA2 encodes a multifunctional carbamoyl phosphate synthetase-aspartate transcarbamylase of 220,000 molecular weight. We determined the nucleotide sequence of the 5' proximal part of the gene which is responsible for the glutamine amide transfer function of the carbamoyl phosphate synthetase activity. Alignment of the enzyme sequence derived from URA2 with sequences from Escherichia coli carA carB and yeast arginine-specific CP A1 CP A2 indicates that monofunctional and bifunctional carbamoyl phosphate synthetases are probably homologous. The URA2-derived enzyme organization is NH2-carbamoyl phosphate synthetase-aspartate transcarbamylase-CO2H. PMID- 3039295 TI - The kdsA gene coding for 3-deoxy-D-manno-octulosonic acid 8-phosphate synthetase is part of an operon in Escherichia coli. AB - The kdsA gene of Escherichia coli was sequenced. It consists of 284 codons and is the last gene of a larger transcription unit. The mRNA is terminated by a rho independent termination signal with the potential to be active in both orientations. This region is followed by another termination signal which seems to be active in the opposite orientation. Upstream of the kdsA gene a second open reading frame (ORF) was found; both this and the kdsA gene are transcribed in the same mRNA molecule when coded from the chromosome. In a plasmid-carried insert transcription starts from a cryptic promoter within the ORF preceding the kdsA gene. This promoter is not active in the intact chromosome. PMID- 3039296 TI - Specificity of bacteriophage Mu excision. AB - To study the excision of bacteriophage Mu at the DNA sequence level, the Mu derived phage lambda placMu3 was transposed to the transcribed but non-translated leader region of a plasmid-borne tetracycline (tet) resistance gene. Revertants (excision products) were then selected by Tet+ restoration of Tet+ and characterized. Of 21 independent Tet+ revertants, 17 contained simple deletions of most or all of lambda placMu3, while the other four contained more complex rearrangements in which one end of lambda placMu3 had been transposed, and most of the prophage had been deleted. The deletion endpoints were found in short direct repeats in each of the complex rearrangements and in 11 of the 17 simple deletion excisants. The results suggest models of slipped mispairing of template and nascent DNA strands facilitated by proteins of the Mu transposition machinery. PMID- 3039297 TI - Isolation of stable aroA mutants of Salmonella typhi Ty2: properties and preliminary characterisation in mice. AB - Derivatives of the Salmonella typhi strain Ty2 carrying stable mutations in the aroA gene were isolated. The mutations were generated by transducing an aroA::Tn10 marker into Ty2 and selecting for derivatives which were tetracycline sensitive and dependent on aromatic compounds for growth. Isolates that did not revert to aromatic compound independence at a detectable frequency were obtained. An S. typhimurium derived aroA specific DNA probe was used to demonstrate the presence of DNA rearrangements in the aroA region of the chromosome of some of the S. typhi aroA mutants. Most of these isolates still expressed Vi antigen. Aromatic compound dependent mutants of S. typhi were less virulent in mice than S. typhi Ty2 following intraperitoneal challenge with bacteria suspended in mucin. Mice immunised with one of these mutants, named WBL85-1, were protected against a potentially lethal challenge of S. typhi Ty2. PMID- 3039298 TI - Analysis of the F plasmid centromere. AB - The nucleotide sequence of the cis-acting partition site (centromere) of the miniF plasmid has been determined. Its most notable feature is a reiterated 43 base pair unit. A series of plasmids deleted for portions of the repeat region was constructed and tested for incompatibility with R386 and for stability of inheritance. The extent of incompatibility with R386 was correlated with the number of repeat units. In contrast, the great majority of the repeats were not needed for miniF stability. An adjacent region of unique sequence was also found to be involved in centromere function. PMID- 3039299 TI - Novel rearrangements of IS30 carrying plasmids leading to the reactivation of gene expression. AB - Unusual DNA rearrangements involving the prokaryote mobile genetic element IS30 have been identified. In order to study the potential mechanisms for the reactivation of a gene after IS element insertion, IS30 was introduced between the lacUV5 promoter and the galK gene of the multicopy plasmid pFR100. In this plasmid terminators of RNA transcription in the sequence of IS30 prevent expression of the galK gene from the lacUV5 promoter. A number of independently isolated mutant plasmids re-expressing the galK gene were studied and shown to be tandemly repeated dimers of the original plasmid. However, the two copies of IS30 were tandemly repeated at one of the original sites of insertion, while at the other site IS30 and three base pairs were missing. The repeated copies of IS30 were separated by two base pairs, the same as those originally flanking the elements. An apparently identical mechanism generated cointegrates between a derivative of plasmid pFD51 carrying IS30 upstream of a promoterless galK gene and a derivative of plasmid pACYC177 carrying IS30 inserted into the beta lactamase gene. This arrangement brought the galK gene under the control of the beta-lactamase promoter of pACYC177. A mechanism involving aborted conservative transposition of IS30 is discussed as a possible route for the generation of these novel cointegrates. In a third experiment we isolated an insert of IS30 which was also two base pairs away from an already resident IS30 element. This insertion of IS30 created a strong promoter of RNA transcription, which has the potential to increase the expression of the transposase in the downstream copy of the element. PMID- 3039300 TI - Processing of TY1 proteins and formation of Ty1 virus-like particles in Saccharomyces cerevisiae. AB - We have analysed functional properties of putative proteins encoded by the yeast transposable element, Ty1, by overexpression of TY genes. High-level expression was achieved by appropriate fusion of a Ty sequence, TY9C, to the yeast ADH1 promoter and transformation of yeast cells with this construction. As shown recently by others (Garfinkel et al. 1985; Mellor et al. 1985c) TY overexpression leads to an increase in particle-bound reverse transcriptase activity and to an intracellular accumulation of virus-like particles (Ty-VLPs). We have used a number of deletions in the second open reading frame (TYB) to identify functional domains required for processing and assembly of Ty proteins. Deletions in the TYB region with homology to acid proteases result in overproduction of an unprocessed form of the TYA protein (pro-TYA) which represents the major protein of Ty-VLPs. One particular mutant construction, TY9C-delta 36, led to the accumulation of a particle-bound, 160 kDa protein which cross-reacted with a mouse antiserum raised against purified pro-TYA protein. This supports the hypothesis that TYB is expressed as a TYA/TYB fusion protein which is processed by a TYB-encoded protease activity. Ty-VLPs are formed in the absence of protein processing and even when the TYB gene is not expressed. Thus, we assume that the assembly of Ty particles occurs prior to processing of Ty proteins. PMID- 3039301 TI - Involvement of ack-pta operon products in alpha-ketobutyrate metabolism by Salmonella typhimurium. AB - The herbicide sulfometuron methyl inhibits acetolactate synthase II of Salmonella typhimurium, resulting in toxic accumulation of alpha-ketobutyrate. Four mutants, containing Tn10 insertions in the acetate kinase (ack) or phosphotransacetylase (pta) genes, were found among a collection of mutants hypersensitive to sulfometuron methyl. The genetic map location of these four Tn10 insertions at 46 min was identical to that of ack and pta point mutants. The insertion and point mutants shared the following phenotypes: resistance to fluoroacetate, sensitivity to alizarin yellow, inability to utilize inositol as a sole carbon source, and hypersensitivity to sulfometuron methyl. Three of the four Tn10 insertion mutants were deficient in phosphotransacetylase but not in acetate kinase activities, indicating insertion of Tn10 in the pta gene. The fourth mutant contained an insertion in the ack gene and was deficient in both acetate kinase and phosphotransacetylase activities. This polarity is consistent with cotranscription of ack and pta. All ack and pta mutants tested were defective in alpha-ketobutyrate turnover. Acetate kinase and phosphotransacetylase are proposed to be part of a pathway for alpha-ketobutyrate metabolism. Propionyl CoA, an intermediate of that pathway, and propionate, the product of the pathway, accumulated upon inhibition of acetolactate synthase. PMID- 3039302 TI - Nucleotide sequence of the kanamycin resistance determinant of the pneumococcal transposon Tn1545: evolutionary relationships and transcriptional analysis of aphA-3 genes. AB - The nucleotide sequence of the kanamycin resistance determinant aphA-3 encoded by transposon Tn1545 from Streptococcus pneumoniae was determined and compared to those of plasmids pJH1 and pIP1433 from Streptococcus faecalis and Campylobacter coli, respectively. The three sequences were found to be identical and differed by two substitutions and the deletion of a codon from that of plasmid pSH2 from Staphylococcus aureus. Comparison of the 5' noncoding sequences indicated that the regions containing the aphA-3 gene in pJH1 and in Tn1545 evolved independently by deletion from a sequence similar to that found in pIP1433. In the latter plasmid, aphA-3 is transcribed from a promoter, P1, which is flanked by two 12-base pair direct repeats. The rearrangement observed in pJH1 removed one of these recombinogenic sites and altered the -10 and 3' flanking sequences of P1. The promoter thus generated. P1', allows expression of similar level of kanamycin resistance as P1. However, fusion experiments carried out with a promotorless chloramphenicol acetyltransferase gene indicated that the canonical promoter P1 is significantly less efficient than P1'. From analysis of the thermodynamic properties of these promoters, we conclude that this difference in strength reflects the melting properties of the -10 sequences. The transition from pIP1433 to pJH1 may correspond to the progression of a molecule structurally unstable to a more stable one combined with the need to maintain an efficient promoter upstream of the aphA-3 gene. The deletion event in Tn1545, which occurred between the two 12-base pair directly repeated sequences, removed P1 in its entirety.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3039303 TI - Cloning and preliminary characterization of the sacS locus from Bacillus subtilis which controls the regulation of the exoenzyme levansucrase. AB - The regulation of sacB, the gene encoding Bacillus subtilis levansucrase is altered by mutations located in several loci unlinked to sacB. Amongst these, the sacS locus seems to play an important role in the induction of sacB by sucrose. We have cloned sacS and found evidence suggesting that it contains two genes. The product of the first gene might repress the expression of the second; the second gene encodes a positive regulator of levansucrase synthesis, since its deletion abolishes this synthesis. There is a palindromic sequence resembling Q independent terminators between the sacB promoter and the structural gene. Mutations affecting this palindrome make sacB constitutive. This suggests that the putative terminator is involved in the induction of sacB by sucrose. We discuss the possibility that the sacS-encoded positive regulator is a sucrose dependent antiterminator which modulates transcription termination between the sacB promoter and the structural gene. PMID- 3039304 TI - Isolation and characterization of mutants of Kluyveromyces lactis defective in lactose transport. AB - Mutants of Kluyveromyces lactis defective in lactose transport were identified among lactose-resistant revertants of lactose-sensitive strains. The mutations are closely linked to the beta-galactosidase gene, LAC4, and they are located in a previously identified gene, LAC12, which has been shown to code for a lactose permease. Our data establish that LAC12 is the only lactose permease gene in K. lactis. The lactose permease also transports galactose. LAC12 is transcribed in a direction opposite to that of LAC4, there being about 2.5 kb between their transcription start sites. Transcription of LAC12 is inducible as is that of all other structural genes in the lactose-galactose regulon of K. lactis. PMID- 3039305 TI - Characterisation of the hydroxystreptomycin phosphotransferase gene (sph) of Streptomyces glaucescens: nucleotide sequence and promoter analysis. AB - The nucleotide sequence of a 1384 bp fragment containing the coding and promoter sequences of the streptomycin phosphotransferase gene (sph) of the hydroxystreptomycin-producing Streptomyces glaucescens was determined. Evidence for an ATG as translation start codon for sph was derived from a comparison with the amino-terminal amino acid sequence of an aminoglycoside phosphotransferase (aphD gene product) of S. griseus, exhibiting a high degree of amino acid homology to the deduced amino acid sequence of the S. glaucescens sph gene product. Transcriptional start and termination sites for the sph gene were identified by primer extension and/or nuclease S1 mapping experiments. The promoter region of the sph gene appears to be complex since tandemly arranged promoters (orfIp1, orfIp2) initiating transcription of a likely coding region (ORFI) in the opposite direction overlap sph promoter sequences. The presumptive sphp and orfIp1 promoters show considerable sequence similarities in the -10 region to Escherichia coli consensus promoter sequences but no homology to E. coli or Streptomyces -35 regions. PMID- 3039306 TI - Gene cluster for streptomycin biosynthesis in Streptomyces griseus: analysis of a central region including the major resistance gene. AB - A central segment of a cluster of biosynthetic genes for the antibiotic streptomycin cloned from Streptomyces griseus was analysed for open reading frames, as well as for transcriptional and translational activity. The nucleotide sequence revealed two significant open reading frames, ORF1 and APH(6), orientated in opposite directions and with a spacer of 885 bp between the start codons. The first, ORF1, had a coding capacity of 38 kDa. One open reading frame, APH(6), was identified as the major resistance gene coding for streptomycin 6 phosphotransferase, a protein of 307 amino acid residues and 33 kDa. Sequence determination of the first 14 N-terminal amino acid residues of the purified APH(6) enzyme protein was in agreement with the proposed primary structure. The possible identity of the presumed gene product of ORF1 with an in vitro translated protein (apparent molecular weight 41 kDa) is discussed. Comparison of the two APH(6) genes from S. griseus and the hydroxystreptomycin-producing S. glaucescens (cf. Vogtli and Hutter 1987) revealed 75% nucleotide sequence homology in the coding region and 74% conservation of the polypeptide sequence. Two protein domains which are highly conserved in other antibiotic and protein phosphotransferases were detected. PMID- 3039307 TI - The origin of transfer (oriT) of the conjugative plasmid R46: characterization by deletion analysis and DNA sequencing. AB - The origin of transfer (oriT) is the sequence within which conjugal transfer of plasmid DNA is initiated, and is absolutely required in cis for plasmid mobilization. We have cloned oriT from the 52 kb IncN plasmid R46 on a 600 bp fragment, and mapped the limits of the relevant sequence by deletion analysis and transposon mutagenesis. The nucleotide sequence of the oriT region contains 13 direct repeats of an 11 bp consensus sequence, 3 different pairs of 10 bp inverted repeats, and a segment that is extremely A-T rich. The direct repeats are within a region required for high frequency transfer and their sequence is such that their periodic alignment along the helix may induce curvature of the DNA. Analysis of Tn1725 insertions within the sequenced fragment of R46 revealed that, unlike most other transposons, transposition of Tn1725 can cause target sequence duplications of three different sizes. PMID- 3039308 TI - Regulation of NAD metabolism in Salmonella typhimurium: genetic analysis and cloning of the nadR repressor locus. AB - The nadR locus (99 min) controls the transcription of several genes involved with either the biosynthesis (nadAB) or recycling (pncB) of NAD in Salmonella typhimurium. Point mutations in this locus were found to cause defects either in the transport of nicotinamide mononucleotide (PnuA-), the regulation of nadAB (NadR-) or both transport and regulation (PnuA-NadR-). Deletions or insertions into nadR always resulted in the PnuA- NadR- phenotypes. Merodiploids constructed with various combinations of PnuA-, NadR- or PnuA- NadR- strains indicate a single complementation group. The results suggest the NadR product is a bifunctional regulatory protein. Operon fusions to lacZ (nadR :: Mud1-8) were used to show that nadR is not autoregulated and is transcribed in a clockwise direction. The gene was also cloned and located within a 2 kb EcoR1-Bg/II fragment. PMID- 3039309 TI - A new family of the poly-deoxyadenylated class of Drosophila transposable elements identified by a representative member at the dunce locus. AB - Two transposable elements have been identified at the dunce locus in chromosomes recovered after a premeiotic and interchromosomal conversion event occurred at this gene. One is approximately 8.2 kb and is inserted near the 5' end of the gene. This element was identified by sequence analysis as a member of the B104 (roo) family of copia-like transposable elements. The second resides near the 3' end of the gene and represents a new family of the class of poly-deoxyadenylated [poly(dA)] transposable elements. It is 0.38 kb in length and has one terminus consisting of a stretch of 29 deoxyadenosine residues with a polyadenylation site like those found in mRNA molecules, located about 20 bp away from the poly(dA) stretch. Fourteen base pairs of genome DNA is duplicated at the target site of this element. PMID- 3039310 TI - Chromosomal initiation in Bacillus subtilis may involve two closely linked origins. AB - When the dnaB37 initiation mutant of Bacillus subtilis is returned to a permissive temperature following a period at 45 degrees C, a synchronous round of DNA replication immediately ensues. Using this system we have been able to analyse the first fragments to be replicated while avoiding the use of thymine starvation or inhibitors of DNA replication. Such treatments are necessary to achieve even modest synchrony in germinating spores. Our results showed that the first fragment to be replicated was a 4 kb BamHI-SalI restriction fragment, BS6. In contrast, when the analysis was performed out in the presence of novobiocin, an inhibitor of DNA gyrase, replication from BS6 was inhibited and the first fragment to be replicated was BS5, a 5.6 kb fragment located 1.7 kb to the right of BS6. Replication from both putative origins was suppressed by rifamycin and was dependent upon dnaB. The results are discussed in relation to previous attempts to identify the first replicating fragment in germinating spores. We also discuss the possibility that B. subtilis contains two origins and suggest that either can act as the primary origin under certain conditions, or alternatively that both origins may act in concert in normal bidirectional replication, each site being required for the leading strand in each direction. PMID- 3039311 TI - The plasmids of Acetobacter xylinum and their interaction with the host chromosome. AB - Acetobacter xylinum contains a complex system of plasmid DNA molecules. Plasmids of molecular weights or copy numbers different from the original wild-type, are found in different types of mutants. Restriction endonuclease digestion and DNA/DNA hybridization analysis, showed that the plasmids often contained partly, but not completely the same DNA sequences. Two of these plasmid classes were analysed in more detail, and could be shown to differ in size by about 5 kb. Hybridization analysis using cloned DNA fragments as probes, showed that sequences lacking in the smallest plasmid were still present in a DNA fraction co migrating with linearized chromosomal DNA. In addition, at least part of the DNA in the smallest plasmid was present both in the plasmid and chromosomal DNA fraction. Analysis of a particular strain containing an insertion of transposon Tn1, also indicated the existence of complex interactions between plasmids and chromosomal DNA. Together with experiments on conjugative transfer and curing of the plasmids, the results indicate that at least part of the genetic system of A. xylinum is unusual when compared to that of other genetically characterized bacteria. PMID- 3039312 TI - Isolation and characterization of an amphipathic antigen from Corynebacterium diphtheriae. AB - Amphipathic antigen was isolated from Corynebacterium diphtheriae Park-Williams number 8 cells by extraction with 47.5% phenol, nuclease treatment and gel filtration on Sepharose 6B. The chemical composition of the amphipathic antigen was hexose (73.8%), pentose (4.6%), fatty acids (9.8%) and glycerol (4.5%). The amphipathic antigen contained arabinose and mannose as sugars at a molar ratio of 1:6 and the major fatty acids were palmitic (C16:0) acid and palmitoleic (C16:1) acid (64.2% and 26.2%, respectively). The amphipathic antigen sensitized sheep erythrocytes and had definite immunobiological activities: viz mitogenic activity on murine splenocytes, stimulatory activity on guinea pig peritoneal macrophages and human complement activation. Deacylation of the amphipathic antigen with alkali treatment lost the sheep erythrocyte-sensitizing ability and some immunobiological activities. The isolated amphipathic antigen described is not a lipoteichoic acid and is different from the teichoic acid of C. diphtheriae. PMID- 3039313 TI - Characteristics of lung pericytes in culture including their growth inhibition by endothelial substrate. AB - Pericytes and endothelial cells from the same sample of adult rat lung have been separately established in culture by use of selective growth media. The endothelial cells are positive and the pericytes negative for angiotensin converting enzyme activity and tissue plasminogen activator. Morphologically in culture, the pericytes are similar to pericytes from bovine retina and other sites and show positive immunofluorescence to both human platelet (non-muscle) myosin and smooth muscle myosin. In this respect they resemble smooth muscle cells grown from the rat main pulmonary artery, but lack the myofilaments and dense bodies characteristic of muscle cells. Lung endothelial cells and fibroblasts are positive only for platelet myosin. Pericytes in culture demonstrate an unusual growth response to endothelial substrate, obtained by removing confluent endothelial monolayers with nonionic detergent or alkali. When plated onto this material at low density, pericyte growth is inhibited. By contrast, the substrate stimulates the growth of endothelial cells and has no effect on smooth muscle cells. Initial attachment of endothelial cells and pericytes to the substrate is similar. PMID- 3039314 TI - Heterogeneity in immunologic functions of rat alveolar macrophages--their accessory cell function and IL-1 production. AB - Alveolar macrophages (AM) from normal rats were separated into 4 different density fractions by centrifugation on a discontinuous Percoll gradient. These fractionated (I-IV) AM, as well as unfractionated (UF) AM, were then tested for their capacities to regulate mitogen-induced T cell proliferation. Concanavalin A (Con A)-induced response of nylon wool-passed non-adherent splenic T lymphocytes was suppressed by addition of UF or higher density (III and IV) AM, while an intermediate density (II) AM fraction could enhance T cell response in a dose dependent manner. Similar effects of UF or fractionated (I-IV) AM on T cell responses were noted when the cultures were exposed in vitro to inert, non fibrogenic titanium dioxide (TiO2) particles. On the contrary, T cell response was sustained by addition of UF or higher density (III and IV) AM, and was also more prominently enhanced by an intermediate density (II) AM after the in vitro exposures to fibrogenic dust particles, like silica and asbestos. Higher interleukin 1 (IL-1) activity was detected from these silica- or asbestos-exposed cultures of UF and fractionated (II, III, and IV) AM. The IL-1 activity was also highly detectable from the cultures of an intermediate density (II) AM fraction when cultures were unexposed or exposed in vitro to TiO2 particles. The Ia antigen expression on the surface of UF or fractionated (II, III, and IV) AM was elevated in the Con A-pulsed co-cultures, but not significantly different whether or not they were exposed in vitro to dust particles. These results may indicate the presence of heterogeneity in accessory cell functions and IL-1 production among rat AM. PMID- 3039315 TI - Interleukin 1 production and accessory cell function of rat alveolar macrophages exposed to mineral dust particles. AB - Immunostimulatory effects of respirable mineral dust particles on alveolar macrophages (AM) and T lymphocytes were tested in vitro. When rat AM were incubated with fibrogenic silica and asbestos particles, a significant interleukin 1 (IL-1) activity was generated into the culture supernatants, whereas neither AM alone nor AM incubated with non-fibrogenic titanium dioxide (TiO2) particles, however, produced a detectable amount of IL-1. Interleukin 2 (IL-2) activity, as tested with IL-2-dependent CTLL-2 assays, was not detectable from all of these cultures. It was also revealed that concanavalin A-induced proliferation of T lymphocytes was enhanced in autologous AM and nylon wool passed spleen cell co-cultures incubated only with fibrogenic particles, but not with non-fibrogenic particles. Higher IL-1 activity was detected only from co cultures exposed to fibrogenic particles, whereas IL-2 activity was almost similar among these co-cultures. These results indicate the differences in immunostimulatory effects on pulmonary macrophages and T lymphocytes among a variety of mineral dust particles. PMID- 3039316 TI - Antigenic and biochemical characterization of poliovirus type 1 isolates. AB - By the introduction of Sabin oral poliovirus vaccine, the circulation of wild type polioviruses has virtually disappeared in Japan. However, an outbreak of poliomyelitis associated with sporadic transmission of type 1 wild strain occurred in Nagano in 1980. Furthermore, we found that some type 1 wild strains were introduced into Japan from abroad in 1981. In recent surveys, the two poliovirus type 1 isolates which have non-vaccine-like antigenic character were detected in Aichi. Then, an investigation to trace the origin of these strains was performed, by using intratypic serodifferentiation and biochemical techniques. Electrophoretic migration patterns of their structural polypeptides were quite different from the vaccine virus. In the oligonucleotide mapping, however, one of them gave patterns very similar to those of the vaccine virus. We could conclude that one originated most probably from wild strains, and the other was an antigenic variant derived from the vaccine virus. It showed that oligonucleotide mapping was a very useful method for identification of antigenic modified Sabin type 1 derivatives. PMID- 3039317 TI - The bactericidal mechanisms of polymorphonuclear leukocytes against Bacteroides fragilis: significance of the oxygen-dependent system. AB - The mechanism by which polymorphonuclear leukocytes (PMNs) kill ingested Bacteroides fragilis was examined using PMNs from patients with chronic granulomatous disease (CGD) which is an inherited disease characterized by the defect of their PMNs in oxygen-radical generation. The phagocytosis of B. fragilis by PMNs from CGD patients was comparable to that by normal PMNs. Although CGD cells killed B. fragilis to some extent, they did so less effectively than the normal PMNS. B. fragilis was killed by a xanthine oxidase system that generates oxygen radicals. When PMNs were incubated with opsonized B. fragilis, B. fragilis triggered the release of O2- and H2O2 from normal PMNs. Thus, normal PMNs appear to kill B. fragilis by both oxygen-dependent and oxygen independent mechanisms. PMID- 3039318 TI - Can lipid peroxidation damage the amputated digit prior to replantation? PMID- 3039319 TI - Asthma and diabetes mellitus: a biochemical basis for antithetical features. AB - Diabetes mellitus and asthma have many antipodal features. Although both are common disorders, concurrence occurs less often than would be predicted. When co existence does occur, the cases are generally mild, and effective treatment of one disease frequently exacerbates the other. The hypothesis is advanced that basilar membrane concentrations of heparan sulfate differ in these two diseases and that this difference may account for the antithetical features. An experimental basis for postulating increased concentrations of extracellular heparan sulfate in asthma and diminished concentrations in diabetes is cited. A rationale for tying these differences to the polar activities of cholinergic transmission and atherogenesis in the two diseases is advanced. Diminished heparan sulfate concentrations in diabetes may down-regulate the transmission of vagal impulses to insulin-producing pancreatic cells, and thereby impair both the continued vitality of these cells, and the acetylcholine modulated potentiation of glucose-induced insulin release. PMID- 3039320 TI - ACTH regulates blood pressure in man. AB - In normal and hypertensive subjects, many stimuli which increase ACTH secretion also increase blood pressure and conversely, many drugs which decrease ACTH secretion also decrease blood pressure. We postulate that the hypothalamopituitary adrenal axis plays a role in blood pressure regulation in normal and hypertensive man. PMID- 3039321 TI - Use of gallium, colloids and pertechnetate as carriers of drugs for selective action. AB - A pharmacodynamic hypothesis is formulated which considers using substances such as gallium, with predilection to locate in neoplasm and inflammation, to increase the therapeutic effect in the disease-affected area. Drugs carried by a colloid can be directed to the liver, IDA derivatives which are metabolized by hepatocytes can also be used as transporters of drugs to the liver and to the hepatobiliary tree because they are eliminated through that route. In diseases which affect the blood-brain barrier substance such as pertechnetate can be used as drug carriers. Pertechnetate and other substances which are eliminated through the kidneys may be used as carriers for drugs in certain renal diseases. This therapeutic approach can be used if the amount of carrier is not toxic to the target organ, if the time of action is sufficient and if the bond between drug and carrier is strong. PMID- 3039322 TI - Anti-receptor antibodies designed to elicit "internal image"-bearing anti idiotypes: a possible AIDS vaccine. AB - Two obstacles hinder the development of an AIDS vaccine: (1) the AIDS virus exhibits extensive amino acid heterogeneity between isolates and (2) antibodies elicited by virus during the course of natural infection are often non neutralizing. A vaccine designed to induce anti-idiotypic antibodies against the virus' receptor on T-cells, T4, should, in principle, overcome these obstacles. Such antibody could contain an "internal image" of T4 and bind the receptor binding domain of the virus. Since this domain is both critical to function and, therefore, poorly susceptible to antigenic variation, anti-receptor anti idiotypic antibodies may demonstrate broad, strain-independent crossreactivity and block viral adherence. PMID- 3039323 TI - The inverse relationship between drug affinity and effectiveness: prediction under rate theory, paradox under occupancy theory. AB - The two major theories of drug action are occupancy theory and rate theory. Although the two theories make a number of predictions that should serve to clearly distinguish one from the other, data generated over the last 25 years in intact muscle or organ systems have failed in this respect because the interpretation of those data has been confounded by unknown and unmeasurable factors such as the rates and extent of drug diffusion, uptake and metabolism. In the present communication rate theory and occupancy theory are discussed in the light of data obtained from a broken-cell system in which uncontrollable factors are minimized, the beta adrenergic-responsive adenylate cyclase of the S49 mouse lymphoma cell line. It is pointed out that rate theory predicts an inverse relationship between the affinity of isoproterenol (INE) for the beta adrenergic receptor and the magnitude of stimulation of adenylate cyclase. On the other hand, occupancy theory predicts that the affinity of INE and the magnitude of its effect will be directly related. By fitting data obtained in this system to the simple equations derived from the two theories it is demonstrated that the available data are in excellent agreement with the predictions of rate theory and diametrically opposed to the predictions of occupancy theory. PMID- 3039324 TI - Barmah Forest virus infections in humans in New South Wales. AB - Antibodies to Barmah Forest virus, a member of the alphavirus group, which was first isolated in 1974, have been found to be widespread in humans in New South Wales. Antibody studies showed a higher prevalence in the north coastal zones of the State, and lower rates in individuals who were living in all other biophysical zones. Antibody rates were significantly higher in male than in female subjects. The pathogenicity of the Barmah Forest virus is at present not known. PMID- 3039325 TI - Health risks from tick-transmitted arboviruses on Australia's Great Barrier Reef. PMID- 3039326 TI - Puzzling changes in cervical cancer in young women. PMID- 3039327 TI - [Neurophysiological study of a group of workers exposed to industrial heptane in a rubber shoe factory]. PMID- 3039328 TI - Ampicillin/sulbactam (Unasyn). PMID- 3039329 TI - [Comparison of isolated (guar) and natural (Musli) dietary fiber in the treatment of type II diabetes]. PMID- 3039330 TI - [Immunologic disorders and viral infections in homozygous beta-thalassemia]. PMID- 3039331 TI - [Congenital pulmonary cystic adenomatosis]. PMID- 3039332 TI - Airway receptors and asthma. AB - Airway caliber is determined by a balance between many constrictor and dilator agents, which bring about their effects on the various target cells of the airway by activating specific cell surface receptors. Asthma and bronchial hyperresponsiveness may be viewed as an imbalance between excitatory and inhibitory receptor-mediated effects on the various target cells in the airway. Airway receptors can be grouped into those mediating the effects of the autonomic nervous system on the airways or those mediating the effects of the various mediators which may be generated in asthma. There is no convincing evidence that a fundamental defect in receptor function is involved in the pathogenesis of asthma, although minor abnormalities have been described. Recently there has been a considerable increase in our understanding of receptor function and control, which should throw light on the pathogenesis of airway obstruction and may lead to advances in asthma therapy. This may also lead to the development of novel drugs for treating asthma in the future. PMID- 3039333 TI - Enterovirus surveillance--United States, 1987. PMID- 3039334 TI - Revision of the CDC surveillance case definition for acquired immunodeficiency syndrome. Council of State and Territorial Epidemiologists; AIDS Program, Center for Infectious Diseases. PMID- 3039335 TI - Beta-adrenergic receptor subtype is an intrinsic property of the receptor gene product. AB - The gene encoding the hamster beta-adrenergic receptor (beta AR) was expressed in mouse C6 glioma cells, a cell line which normally expressed the beta 1 subtype of the receptor. Upon transfection with the hamster beta AR gene, the cells expressed increased levels of beta AR, as assessed both by protein immunoblotting and by the binding of the radiolabeled antagonist 125I-cyanopindolol. This newly expressed receptor was of the beta 2 subtype, as determined with a variety of agonists and antagonists. These results suggest that the subtype of the beta AR is an intrinsic property of the gene product and is not the result of a post translational modification of the receptor by the cell in which it is expressed. PMID- 3039336 TI - Immuno cross-reactivity suggests that catecholamine biosynthesis enzymes and beta adrenergic receptors may be related. AB - Turkey red blood cell, beta 1-adrenergic receptors (BARs) were prepared to electrophoretic homogeneity by affinity chromatography, size exclusion high performance liquid chromatography, and preparative sodium dodecyl sulfate polyacrylamide gel electrophoresis and used to prepare rabbit polyclonal anti-BAR antibodies. Anti-BAR activity was confirmed by immunoadsorption of [125I]cyanopindolol-labeled BAR to a protein A affinity column using the anti-BAR antibodies. BAR was compared to the catecholamine biosynthetic enzyme dopamine B hydroxylase (DBH) by anti-BAR antibody cross-reactivity. DBH was purified from bovine adrenal medullae chromaffin vesicles by ion exchange, size exclusion, and concanavalin A-Sepharose chromatography. Final DBH specific activities were 42 +/ 4 units/mg of protein. Homogeneity was confirmed by nondenaturing polyacrylamide gel electrophoresis. Both DBH and BAR were recognized by the anti-BAR antibodies on Western transfer and immunoblotting. No interactions were observed with preimmune controls. Similar results were obtained with glycosylated and deglycosylated DBH, suggesting that the anti-BAR antibodies recognize specific portions of DBH amino acid sequence and not associated carbohydrate. DBH-cross reactive antibodies were also purified by affinity chromatography using immobilized DBH and shown to immunoadsorb [125I]cyanopindolol-labeled BAR by protein A affinity chromatography. These results suggest that the catecholamine biosynthetic enzyme DBH and BAR may be related in structure. PMID- 3039337 TI - Relative abundance of two molecular forms of Na+,K+-ATPase in the ferret heart: developmental changes and associated alterations of digitalis sensitivity. AB - We have previously reported that adult ferret hearts have high affinity and low affinity ouabain-binding sites in approximately equal abundance corresponding, respectively, to alpha(+) and alpha subunits of Na+,K+-ATPase. In the present study, possible changes in relative abundance of the two molecular forms of Na+,K+-ATPase during postnatal development were examined. In 5-day-old or 7- to 10-day-old ferret heart, there was a predominance of the alpha form of Na+,K+ ATPase, the alpha(+)/alpha ratio being approximately 1:7 and 1:4 at 5 and 7-10 days, respectively, in contrast to a 1:1 ratio observed previously in adult heart. Corresponding to the reduced number of high affinity ouabain-binding sites associated with the alpha(+) form of Na+,K+-ATPase, ouabain sensitivity of the Na+,K+-ATPase isolated from newborn ferret heart was lower than that of adult ferret heart. Furthermore, the low dose positive inotropic effect, which was apparent in the adult ferret heart, could not be observed in the newborn ferret heart. The present finding may represent a plausible explanation for low digitalis sensitivity of neonatal heart observed in several mammalian species. PMID- 3039338 TI - SKF 104353, a high affinity antagonist for human and guinea pig lung leukotriene D4 receptor, blocked phosphatidylinositol metabolism and thromboxane synthesis induced by leukotriene D4. AB - SKF 104353 (2(S)-hydroxyl-3(R)-carboxyethylthio)-3-[2-(8-phenyloctyl) phenyl] propanoic acid) is a synthetic structural analog of leukotrienes D4 and E4 (LTD4, LTE4). This compound binds to guinea pig and human lung LTD4 receptors with affinities (Ki) of 5 +/- 2 and 10 +/- 3 nM, respectively. The Ki values of a reference compound, FPL 55712, were 2200 and 4500 nM, respectively, approximately 400- and 500-fold less effective than SKF 104353. LTD4- and LTE4-induced biosynthesis of thromboxane B2 has been shown to be mediated by LTD4 receptors in guinea pig lung in vitro. SKF 104353 did not induce synthesis of TxB2 in this system at concentrations of 1-20 microM. When SKF 104353 and increasing concentrations of LTD4 were incubated with guinea pig lung, the dose response curve of LTD4-induced TxB2 biosynthesis was shifted to the right with a -log[KB] = 8.4 +/- 0.2. LTD4-induced phosphatidylinositol (PI) hydrolysis in guinea pig lung has been shown to be the major signal transduction mechanism. In this system, SKF 104353 (1-20 microM) did not promote PI hydrolysis. Pretreatment of the [3H]myo-inositol-labeled guinea pig lung with SKF 104353 shifted the LTD4 induced PI hydrolysis dose response curve to the right, indicating that SKF 104353 inhibited LTD4 receptor-mediated intracellular second messenger formation. These results demonstrate that SKF 104353 is a high affinity, specific LTD4 receptor antagonist. It inhibited LTD4-induced PI hydrolysis and TxB2 biosynthesis in guinea pig lung. SKF 104353 may prove to be an important research tool for research on the activities of leukotrienes and of value therapeutically in the treatment of leukotriene-mediated diseases. PMID- 3039339 TI - Heterologous regulations of cAMP responses in pregnant rat myometrium. Evolution from a stimulatory to an inhibitory prostaglandin E2 and prostacyclin effect. AB - The regulation of cAMP generation in rat myometrium during gestation was investigated comparatively to the stimulations evoked in the estrogen-dominated myometrium. At the onset of gestation, there was a progressive attenuation in both tissue cAMP accumulation and adenylate cyclase activation in response to isoproterenol and prostaglandins (PGs) (PGE2 and prostacyclin, PGI2), as well as to cholera toxin and to forskolin. Minimal responses were observed at midgestation (day 12). The decline in isoproterenol-mediated cAMP accumulation could not be ascribed to alterations in either beta-adrenergic receptor density or coupling properties. Treatment of day 12 myometrium with islet-activating protein (IAP), pertussis toxin, caused a reversal of the attenuated beta adrenergic--as well as cholera toxin--responses, suggesting that the inhibitory regulatory protein, Gi, was involved. However IAP treatment did not improve the PGE2-mediated stimulatory effect. In membranes from day 12 myometrium, the amount of [32P]NAD incorporated by IAP into the Mr = 41,000 peptides (alpha i, subunit of Gi was markedly increased compared to membranes from day 0 tissue. There was also a consistent decrease (25%) of the label incorporated by cholera toxin into the Mr = 52,000 (major) and 45,000/53,000 (minor) peptides (alpha s, subunit of Gs). The advanced stages of gestation were associated with a progressive restoration of cAMP responses to isoproterenol, cholera toxin, and forskolin with a full responsiveness before parturition (day 22). By contrast, the inability of PGE2 to generate any active Gs (stimulatory regulatory protein)-C complex persisted and coincided with the development of PGE2-induced inhibition of cAMP generation due to isoproterenol. The inhibitory effect, which was similarly evoked by PGE2 as well as by PGI2 and PGF2 alpha, was prevented by IAP. The data suggest that alterations of the stimulatory Gs-mediated pathway, which culminated at midgestation, is due at least in part to an increase in the abundance of Gi relative to Gs. Additional alterations operate at the level of the prostaglandin receptor(s) with a conversion from a stimulatory (Gs-mediated) cAMP response in the estrogen-dominated myometrium to an inhibitory (Gi-mediated) effect, fully expressed in the final stage of gestation. PMID- 3039340 TI - Compartmentation of alpha 2-adrenergic receptors in human erythroleukemia (HEL) cells. AB - We have identified alpha 2-adrenergic receptors on human erythroleukemia (HEL) cells, a suspension-grown cell line related to human platelets. properties of receptors were assessed in intact cells by binding of the antagonist [3H]yohimbine and by inhibition of cAMP accumulation. [3H]Yohimbine labeled 5900 +/- 2100 receptors/cell with a Kd of 3.6 +/- 0.9 nM (n = 7). alpha 2-Adrenergic receptors were potently coupled to inhibition of adenylate cyclase, with EC50 values for epinephrine, UK-14,304, and p-aminoclonidine in the low nM range. Treatment of cells with pertussis toxin abolished this response. In radioligand binding studies with membrane preparations [3H]yohimbine and [3H]UK-14,304 bound to the same number of sites (71 versus 69 fmol/mg of protein), and epinephrine competed for [3H]yohimbine binding in a biphasic manner. After addition of GTP, no high affinity [3H]UK-14,304 binding was detected, and epinephrine competed for [3H]yohimbine binding with lower affinity at both 4 degrees and 37 degrees. In studies with intact cells, we detected no specific binding of [3H]UK-14,304 at either 37 degrees or 4 degrees. At 37 degrees, epinephrine competed for all [3H]yohimbine binding sites with a low apparent affinity (Ki = 21 microM), whereas at 4 degrees epinephrine (up to 1 mM) was able to compete for only 59 +/- 13% of [3H]yohimbine-binding sites. The potency of epinephrine in competing for [3H]yohimbine sites in intact cells at 4 degrees was greater than at 37 degrees (Ki = 1 microM) and was similar to that observed with membranes in the presence of GTP. We hypothesize that sites not detectable by epinephrine at 4 degrees are sequestered within the cell. Treatment of HEL cells with pertussis toxin reduced the proportion of receptors on the surface from 51 +/- 12% to 23 +/- 7% (n = 3, p less than 0.05) of the total sites. Treatment of HEL cells with epinephrine (100 microM, 1 hr) reduced the cell surface component to 25 +/- 8% (n = 3) of the total sites. This treatment was not accompanied by significant desensitization of the ability of epinephrine to inhibit cAMP accumulation. We conclude that alpha 2 adrenergic receptors exist in more than one compartment in HEL cells and that interaction of receptors with a guanine nucleotide-binding protein or with agonist may regulate this compartmentation. These cells provide a new model system for the study of expression and metabolism of alpha 2-adrenergic receptors. PMID- 3039341 TI - gamma-Aminobutyric acid receptor binding in fresh mouse brain membranes at 22 degrees C: ligand-induced changes in affinity. AB - Binding of [3H]muscimol to mouse brain gamma-aminobutyric acid receptors has been assayed under more physiological conditions (never-frozen membranes, 22 degrees) than in previous studies (0-4 degrees on frozen-thawed membranes). Binding affinities were lower and agreed more closely with physiological dose response curves. In addition, heterogeneity in affinity was still present, apparently due to a mixture of non-interconvertible subpopulations and ligand-induced changes in kinetics. Super-high affinity sites (Kd less than 10 nM) observed in frozen membranes were not observed in fresh membranes under equilibrium binding conditions, and high affinity sites (Kd congruent to 25 nM) seen at 0 degree had lower affinity (Kd congruent to 250 nM) at 22 degrees. Despite an apparent best fit single-component Scatchard plot for the latter data, cold ligand displacement and association and dissociation rates demonstrated heterogeneity of affinities. Pentobarbital greatly increased the amount of high affinity sites at the expense of low affinity sites in equilibrium binding at 0 degree and slightly increased affinity at 22 degrees. The kinetics of [3H]muscimol association at 22 degrees (kapp = 1-2/min) were virtually independent of the ligand concentration, suggesting either negative cooperativity or an agonist binding-dependent receptor isomerization. Dissociation triggered by addition of excess cold ligand to membranes equilibrated with [3H]muscimol at different concentrations revealed two off-rates with t1/2 values of under 10 sec and 1-3 min; unexpectedly, the ratio of these two populations did not vary with ligand concentration but was constant at 50:50. Dissociation triggered by infinite dilution gave two off-rates, but the slowest component had a t1/2 of about 20 min; including cold muscimol in the dilution buffer increased the off-rate toward that observed in the excess cold ligand method (t1/2 = 1-2 min). The very slow off-rate was only observed following removal of agonist from a previously occupied receptor and suggests receptor isomerization into a high affinity state; this state is similar to that observed at 0 degree. Pentobarbital also favored the production of the receptor state showing very slow [3H]muscimol dissociation upon infinite dilution, opposing the action of high receptor occupancy with cold muscimol. Thus, [3H]muscimol-binding sites observed at 22 degrees with fresh membranes do not show artificially high affinity and have a Kd of 0.2-0.3 microM, closer to the EC50 for chloride channel activation (approximately 2-3 microM and 0.4 microM with pentobarbital).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3039342 TI - Voltage-dependent nitrendipine binding to cardiac sarcolemmal vesicles. AB - The binding of the calcium channel antagonist [3H]nitrendipine (3[H]Ntd) to purified bovine cardiac sarcolemmal vesicles was examined at different membrane potentials. The vesicles were loaded with 150 mM K and exposed to a range of external K concentrations to induce negative internal membrane potentials using N methyl-D-glucamine or choline to substitute for K. The lipophilic cation [3H]tetraphenylphosphonium was used to quantitate the membrane potential, and the vesicles were capable of maintaining a relatively stable negative inside membrane potential for at least 15 min. Equilibrium binding studies of [3H]Ntd performed on vesicles maintained at a depolarized potential (0 mV) revealed 24.7 +/- 5.6% more high affinity [3H]Ntd-binding sites than were present on vesicles maintained at a hyperpolarized potential (approximately -40 to -50 mV) with N-methyl-D glucamine substituting for K. There was no significant change in Kd values which were 0.398 +/- 0.069 and 0.388 +/- 0.032 nM, respectively. When choline was used to substitute for K, comparable results were obtained with 19.01 +/- 3.4% more high affinity [3H]Ntd-binding sites under depolarized conditions and no significant change in Kd (0.356 +/- 0.025 and 0.379 +/- 0.030 nM, respectively). Varying the external K concentration resulted in a graded change in [3H]Ntd binding over the same range of external K concentrations which resulted in the greatest change in membrane potential. These findings suggest that the observed effect was due to changes in membrane potential rather than changes in the composition of the incubation medium. Additional control experiments employing choline-loaded vesicles also support the hypothesis that the observed effect was primarily due to changes in membrane potential. The present results are compared with those from electrophysiological studies. PMID- 3039343 TI - Potent activity of 5-fluoro-2'-deoxyuridine and related compounds against thymidine kinase-deficient (TK-) herpes simplex virus: targeted at thymidylate synthase. AB - 5-Fluorouracil, 5-fluorouridine (FUrd), 5-fluoro-2'-deoxyuridine (FdUrd), 5 fluorocytidine (FCyd), 5-fluoro-2'-deoxycytidine (FdCyd), 5-trifluoro-2' deoxythymidine (F3dThd), and the 5'-monophosphates and 3',5'-cyclic monophosphates thereof were found to inhibit thymidine kinase-deficient (TK-) mutant strains of herpes simplex virus (HSV) at a much lower concentration than the wild-type (TK+) HSV strains. Other 5-substituted 2'-deoxyuridines that have previously been recognized as potent thymidylate synthase inhibitors behaved in a similar fashion. The activity of FdUrd, FdCyd, F3dThd, and their 3',5'-cyclic monophosphates against TK-HSV was readily reversed by 2'-deoxythymidine (dThd) but not by 2'-deoxyuridine (dUrd). These compounds also inhibited the incorporation of [6-3H]dUrd into DNA at a concentration which was up to 5 orders of magnitude lower than the concentration at which the incorporation of [methyl 3H] dThd was inhibited. Thus, while not being a target for the well established anti-HSV compounds in TK+HSV-infected cells, thymidylate synthase appears to be an important target in TK-HSV-infected cells. In addition to dTMP synthase, TK HSV-infected cells appear to reveal other therapeutically exploitable targets such as OMP decarboxylase (towards pyrazofurin), CTP synthase (towards carbodine and its cyclopentenyl analogue), dihydrofolate reductase (towards methotrexate), and S-adenosylhomocysteine hydrolase (towards neplanocins). PMID- 3039344 TI - An Epstein-Barr virus transforming protein associates with vimentin in lymphocytes. AB - The Epstein-Barr virus (EBV) latent infection membrane protein (LMP) is likely to be an important mediator of EBV-induced cell proliferation, since it is one of the few proteins encoded by the virus in latent infection and since production of this protein in Rat-1 cells results in their conversion to a fully transformed phenotype. LMP was previously noted to localize to patches at the cell periphery. In this paper we examine the basis of LMP patching in EBV-infected, transformed lymphocytes. Our data indicate that LMP is associated with the cytoskeletal protein vimentin. Although LMP is fully soluble in isotonic Triton X-100 buffer, only 50% of it is extracted from cells in this solution. The rest remains bound to the cytoskeleton. LMP undergoes phosphorylation, and phosphorylated LMP is preferentially associated with the cytoskeleton. As judged by both immunofluorescence and immunoelectron microscopy, the vimentin network in EBV transformed lymphocytes or EBV-infected Burkitt tumor lymphocytes is abnormal. Vimentin and LMP often colocalize in a single patch near the plasma membrane. In response to Colcemid treatment of EBV-infected cells, vimentin reorganizes into perinuclear rings, as it does in uninfected cells. LMP is associated with these perinuclear rings. Vimentin (or a vimentin-associated protein) may be a transducer of an LMP transmembrane effect in lymphoproliferation. PMID- 3039345 TI - Functional organization of the Aspergillus nidulans trpC promoter. AB - We investigated the functional organization of the Aspergillus nidulans trpC promoter by the sequential removal of sequences upstream of the major trpC mRNA cap site (+1). DNA fragments containing promoter mutations were fused to the Escherichia coli lacZ gene, and a novel method was used to select for integration of the fusion gene at the Aspergillus argB locus. beta-Galactosidase assays and S1 nuclease protection experiments demonstrated that the promoter mutations affected gene expression in three ways: (i) 5' deletions up to -82 resulted in variable increases in beta-galactosidase activity, depending on the growth conditions; (ii) a deletion from -67 to -11 did not alter the level of beta galactosidase activity, but did give rise to mRNAs with aberrant 5' ends; and (iii) a 5' deletion with an endpoint at -11 and an internal deletion from -142 to -11 abolished gene expression. These results indicate that sequences upstream of 82 reduce transcription of the trpC gene and that distinct DNA sequence elements are required for expression versus correct initiation of transcription of the trpC gene. The sequences essential for trpC expression do not include the common eucaryotic promoter elements CCAAT and TATAAA. To our knowledge, this is the first functional analysis of a promoter from a fungus other than Saccharomyces cerevisiae. PMID- 3039346 TI - Synthesis of membrane-bound colony-stimulating factor 1 (CSF-1) and downmodulation of CSF-1 receptors in NIH 3T3 cells transformed by cotransfection of the human CSF-1 and c-fms (CSF-1 receptor) genes. AB - NIH 3T3 cells cotransfected with the human c-fms proto-oncogene together with a 1.6-kilobase cDNA clone encoding a 256-amino-acid precursor of the human mononuclear phagocyte colony-stimulating factor CSF-1 (M-CSF) undergo transformation by an autocrine mechanism. The number of CSF-1 receptors on the surface of transformed cells was regulated by ligand-induced receptor degradation and was inversely proportional to the quantity of CSF-1 produced. A tyrosine-to phenylalanine mutation at position 969 near the receptor carboxyl terminus potentiated its transforming efficiency in cells cotransfected by the CSF-1 gene but did not affect receptor downmodulation. CSF-1 was synthesized as an integral transmembrane glycoprotein that was rapidly dimerized through disulfide bonds. The homodimer was externalized at the cell surface, where it underwent proteolysis to yield the soluble growth factor. Trypsin treatment of viable cells cleaved the plasma membrane form of CSF-1 to molecules of a size indistinguishable from that of the extracellular growth factor, suggesting that trypsinlike proteases regulate the rate of CSF-1 release from transformed cells. The data raise the possibility that this form of membrane-bound CSF-1 might stimulate receptors on adjacent cells through direct cell-cell interactions. PMID- 3039347 TI - Location of sequences within rotavirus SA11 glycoprotein VP7 which direct it to the endoplasmic reticulum. AB - The Simian 11 rotavirus glycoprotein VP7 is directed to the endoplasmic reticulum (ER) of the cell and retained as an integral membrane protein. The gene coding for VP7 predicts two potential initiation codons, each of which precedes a hydrophobic region of amino acids (H1 and H2) with the characteristics of a signal peptide. Using the techniques of gene mutagenesis and expression, we have determined that either hydrophobic domain alone can direct VP7 to the ER. A protein lacking both hydrophobic regions was not transported to the ER. Some polypeptides were directed across the ER membrane and then into the secretory pathway of the cell. For a variant retaining only the H1 domain, secretion was cleavage dependent, since an amino acid change which prevented cleavage also stopped secretion. However, secretion of two other deletion mutants lacking H1 and expressing truncated H2 domains was unaffected by this mutation, suggesting that these proteins were secreted without cleavage of their NH2-terminal hydrophobic regions or secreted after cleavage at a site(s) not predicted by current knowledge. PMID- 3039348 TI - Molecular cloning and genetic analysis of the suppressor-of-white-apricot locus from Drosophila melanogaster. AB - We report genetic and molecular analysis of the suppressor-of-white-apricot [su(wa)] locus, one of several retrotransposon insertion allele-specific suppressor loci in Drosophila melanogaster. First, we isolated and characterized eight new mutations allelic to the original su(wa)1 mutation. These studies demonstrated that su(wa) mutations allelic to su(wa)1 affected a conventional D. melanogaster complementation group. Second, we cloned the chromosomal region containing the su(wa) complementation group by P element transposon tagging. The ca. 14-kilobase region surrounding the su(wa) complementation group contained five distinct transcription units, each with a different developmentally programmed pattern of expression. Third, we used a modified procedure for P mediated gene transfer to identify the transcription unit corresponding to su(wa) by gene transfer. Fourth, we found that the presumptive su(wa) transcription unit produced a family of transcripts (ranging from ca. 3.5 to ca. 5.2 kilobases) in all developmental stages, tissue fractions, and cell lines we examined, suggesting that the gene is universally expressed. PMID- 3039349 TI - Rapid and selective alterations in the expression of cellular genes accompany conditional transcription of Ha-v-ras in NIH 3T3 cells. AB - Hormone treatment of NIH 3T3 cells that contain recombinant fusions between the mouse mammary virus long terminal repeat and the v-ras gene of Harvey murine sarcoma virus results in conditional expression of the ras p21 gene product. Levels of ras mRNA and p21 are maximal after 2 to 4 h of hormone treatment. Analysis of cellular RNA by Northern blotting and nuclease S1 protection assays indicates that the expression of two cellular RNA species increases with kinetics similar to v-ras: v-sis-related RNA and retrovirus-related VL30 RNA. Run-on transcription in isolated nuclei shows that the increase in v-sis-related RNA is not dependent on transcription and therefore must arise by a post-transcriptional mechanism. The increase in VL30 expression is a transcriptional effect. Hormone treatment of normal NIH 3T3 cells has no effect on the expression of these DNA sequences. These results suggest that v-ras stimulation of autocrine factors may play a role in transformation of cells by this gene and also suggest a reverse genetic strategy to determine the nucleic acid sequences and cellular factors involved in the regulation of gene expression that is observed. PMID- 3039350 TI - Negative regulation contributes to tissue specificity of the immunoglobulin heavy chain enhancer. AB - We have identified in and around the immunoglobulin heavy-chain enhancer two apparently distinct negative regulatory elements which repress immunoglobulin H enhancer, simian virus 40 enhancer, and heterologous promoter activity in fibroblasts but not in myeloma cells. We propose that in nonlymphoid cells, negative regulatory elements prevent activation of the immunoglobulin H enhancer by ubiquitous stimulatory trans-acting factors. PMID- 3039351 TI - Human HER2 (neu) promoter: evidence for multiple mechanisms for transcriptional initiation. AB - We localized the 5' region of the human gene HER2 in a cloned fragment of genomic DNA. This clone contained exons 1 to 4 of HER2, spanning the coding sequence for the first 191 amino acids. The promoter region of HER2 was identified upstream to exon 1 by nuclease S1 mapping and by a functional assay in which the promoter region drives the expression of a chloramphenicol acetyltransferase gene. The HER2 promoter is different from the promoter of the epidermal growth factor receptor gene (HER1), and the GC boxes which are typical of the promoter of the epidermal growth factor receptor gene are absent from the HER2 promoter. One major and two minor RNA start sites located at nucleotides 178, 244, and 257 upstream to the initiator ATG were identified. The first one is 21 and 70 base pairs downstream from typical TATAA and CAAT boxes, respectively. This indicates that transcription of HER2/neu can be regulated by a mechanism involving a TATA box, as well as by other unidentified regulatory elements. PMID- 3039352 TI - Abelson virus transformation of an interleukin 2-dependent antigen-specific T cell line. AB - Abelson murine leukemia virus (A-MuLV) carries the gene v-abl, one of a group of oncogenes with structural and functional (tyrosine kinase) homology to three growth factor receptors. Work in this and other laboratories has shown that A MuLV infection can render myeloid and lymphoid cells independent of the growth factors interleukin 3 and granulocyte-macrophage colony-stimulating factor. We have now shown that v-abl can also relieve interleukin 2 (IL-2) dependence in T cells. We infected a cloned IL-2-dependent antigen-specific cell line. Transformed cells were generated which were factor independent and tumorigenic. The transformants each bore unique v-abl DNA inserts and expressed v-abl mRNA. No elevation of expression of either IL-2 or its receptor could be detected in these cells. Thus, A-MuLV can short-circuit the dependence of hematopoietic cells on IL 2, IL-3, and possibly granulocyte-macrophage colony-stimulating factor, none of whose receptors are known to be of the tyrosine kinase type. PMID- 3039353 TI - Unusual sequence element found at the end of an amplicon. AB - In a polyomavirus-transformed rat cell line, designated LPT, the polyomavirus DNA is integrated into a single chromosomal site. Treatment of LPT cells with carcinogens induces amplification of the integrated virus DNA and flanking cellular sequences. We show that the amplification is arrested within a specific cell DNA segment that maps 1.3 to 1.85 kilobases beyond one virus-cell DNA junction, defined as the left junction. We also present the sequence of an 897 base-pair fragment spanning the arrest site. This fragment contains an unusual sequence element, which consists of two contiguous components, a potential cruciform with stems of 6 base pairs and a d(G-A)27 X d(T-C)27 tract, and maps 1,497 to 1,564 nucleotides beyond the left junction. The possibility that this unusual sequence plays a role in the arrest of the amplification process is discussed. PMID- 3039354 TI - Transcriptional inactivation of c-myc and the transferrin receptor in dibutyryl cyclic AMP-treated HL-60 cells. AB - Treatment of HL-60 cells with dibutyryl cyclic AMP induced rapid transcriptional inactivation of c-myc and the transferrin receptor. Transcriptional inactivation was followed by loss of c-myc and transferrin receptor mRNA and protein. Treated cells completed one round of proliferation, followed by growth arrest, G1 synchronization, and monocytic differentiation. These data suggest that cyclic AMP-mediated control of growth and differentiation may be achieved, at least in part, by transcriptional regulation of certain growth-associated proto-oncogenes and growth factor receptor genes. PMID- 3039355 TI - Establishment of a human T-cell hybridoma that produces human macrophage activating factor for superoxide production and translation of messenger RNA of the factor in Xenopus laevis oocyte. AB - Human monoblast-like histiocytic lymphoma cell line U937 was induced by a macrophage activating factor for O2- production (MAF-O) to produce O2- in response to phorbol myristate acetate stimulation. A MAF-O-producing human T-cell hybridoma, F4-29-4, was established which was also found to produce macrophage activating factor for glucose consumption (MAF-G) and colony stimulating factor (CSF) when assayed against mouse bone marrow cells. MAF-O could be successfully separated from CSF but not from MAF-G by phenyl-Sepharose CL-4B chromatography (Phenyl-EG-fraction). To differentiate MAF-O from MAF-G and to explore a route for large scale production of MAF-O and its structural elucidation, total messenger RNA was extracted from a human T-cell hybridoma, clone F4-29-4. This messenger RNA was fractionated on 5-30% sucrose gradient and each fraction was microinjected into Xenopus laevis oocytes. MAF-O activity was found in the supernatant of oocytes injected with messenger RNA sedimentated at about 13.0S, while MAF-G messenger RNA was found to be about 10.5S. The MAF-O activity, synthesized from the injected messenger RNA, was not neutralized with an excess amount of anti-human IFN-gamma anti-serum, suggesting that MAF-O is antigenically different from human IFN-gamma. PMID- 3039356 TI - [Transposition of the Tn5 and Tn10 elements and duplications in Escherichia coli K12 genome]. AB - The kinetics of accumulation of resident transposon copies in a dividing population has been defined using a special experimental system. Analysis of the kinetics made it possible to estimate the probability of transposition for Tn5 as 2.5 X 10(-4) and for Tn10 as 2.3 X 10(-6) per cell per generation. Transposition of the composite elements does not depend on RecBC or RecF pathways of recombination. The fraction of the bacterial population with tandem duplications in the proA region of the genome is permanent for Escherichia coli. It is independent of the recombination pathways (RecBC of RecF) and the integrity of DNA polymerase I. PMID- 3039357 TI - [DNA duplications detect the hidden transposition of the Tn5 element in the Escherichia coli K12 genome]. AB - The frequency of Tn5 transposition localized in an arm of a tandem duplication was estimated as 1.3 X 10(-2) per cell per generation, two orders of magnitude higher than usual one. Approximately thirty per cent of all transpositions usually registered occur from the spontaneous duplications. The effect revealing latent transpositions is in good accordance with a conservative transposition model permitting some interesting predictions: 1. Composite transposons can be a reason for the double stranded cuts in DNA. 2. The transposition frequency in cis for composite elements seems to be many times higher than in trans. 3. Partially transpositions in cis can be recA dependent. 4. The estimation of Tn5 transposition in cis presented in the paper is a minimal one. PMID- 3039358 TI - [Preservation of the beta-galactosidase activity in E. coli cells containing the recombinant pUC19 plasmid]. AB - Two BglII fragments of pJC703 cosmid were inserted into the plasmid PUC19. E. coli cells containing the recombinant PUC19/A plasmid acquired rifampicin resistance due to inserted rpoB gene but still expressed the Lac+ phenotype. PMID- 3039359 TI - [Expression of the pectate lyase gene from Erwinia chrysanthemi ENA49 in cells of other Erwinia species]. AB - The gene for a pectate lyase of E. chrysanthemi ENA49 cloned in a recombinant plasmid pPTL1 (a derivative of RSF1010) was transferred into E. carotovora. The pectate lyase determined by the cloned gene was secreted into the cultural medium from the cells of E. crysanthemi EC16. Partial secretion of the enzyme was registered for E. carotovora cells. The major part of EC1 E. chrysanthemi pectate lyase synthesized by E. carotovora cells is accumulated in periplasmic and cytoplasmic fractions. The obtained results suggest the different specificity or efficiency of pectate lyase secretion systems in the studied Erwinia strains. PMID- 3039360 TI - [Rotaviruses: structural and functional organization of the virion]. AB - The data on the structural and functional organization of human rotaviral virion and rotaviral virions of different species of animals obtained by the native and foreign researchers for the last five years of study have been summarized. The virion ultrastructure, the genomic structure and polypeptide composition of rotaviruses are discussed. The data are presented on the transcription and molecular mapping of rotaviral genome. Special attention is paid to localization of rotaviral proteins in the virion capsid, to their function and derivation. The data on the role of endogenous enzymes in rotaviral transcription and replication are presented separately. PMID- 3039361 TI - [Homology between the ribosomal and RNA-polymerase genes of E. coli and vaccine strain E of Rickettsia prowazekii]. AB - The pIB52 plasmid contains the ribosomal rp1A, J, K, L and RNA-polymerase genes rpo C, B of Escherichia coli. No homology has been found between the plasmid used as a molecular probe and the chromosomal DNA from Rickettsia provazekii in the blot hybridization experiments. PMID- 3039362 TI - [Adaptation of hepatitis A virus strain (JaM-55) originally pathogenic for monkeys to a cell culture]. AB - The results of adaptation of hepatitis A viral strain JaM-55 to the culture of embryo kidney cells FRhk-4 from macaque Rhesus are presented. The viral strain was isolated from a M. fascicularis suffering from spontaneous hepatitis. Before inoculating the cell culture the virus was passaged twice in the M. arctoides capable of reproducing hepatitis. FRhk-4 cell line inoculation by the monkey liver extract, containing the strain HAV-YaM-55, resulted in isolation of single viral particles of hepatitis A in the preparations obtained at the first 3 passages by the 28-31 day of cultivation. Beginning from the fourth passage the abrupt increase in the number of viral particles and hepatitis A antigen was registered. There were no traces of cytopathogenic effect at any level of viral passages in the inoculated cell culture. The adapted virus contains hepatitis A viral RNA identified by spot hybridization with the cloned cDNA of hepatitis A virus. PMID- 3039363 TI - The relationship between radiation-induced and transposon-induced genetic damage during Drosophila spermatogenesis. AB - The combined effect of transposon mobility and X-rays on X-linked recessive lethals and dominant lethals was measured in the germ line of F1 male hybrids in the P-M system of hybrid dysgenesis. X-Linked lethal mutation rate was measured in the chromosome derived from the P-strain father of the M X P cross. Mutations induced in irradiated dysgenic males were compared to those of unirradiated males, as well as to irradiated nondysgenic males derived from M X M crosses. Three four-day broods of sperm were tested for both X-linked lethals and dominant lethals. X-Linked lethal mutation rate in dysgenic control males was 6.38%, 6.36% and 4.55% in broods 1, 2 and 3 respectively, thus showing a decrease in older males. The mutation rate in the same broods of irradiated, nondysgenic control males was 3.66%, 4.46% and 6.38%, respectively. The rate obtained in dysgenic irradiated males was 10.33, 11.16 and 7.97 in the same 3 broods. These results demonstrate that when X-rays and P element mobility were combined as a source of mutagenesis, a strictly additive effect on genetic damage was observed in the first two broods of sperm which represent primarily mature sperm and spermatids respectively. The third brood, representing mostly spermatocytes showed a less than additive effect, probably due to germinal selection. In contrast, the induction of dominant lethals showed a clearly synergistic effect in the last two broods of sperm tested, when X-rays and transposon mobility were combined. The X ray component of dominant lethality in brood 1, representing mostly mature spermatozoa, was negative, indicating a lower than expected lethality induced by X-irradiation in the presence of P element mobility. The X-ray-induced component of dominant lethality, was expressed as the per cent of embryo lethality after adjusting the results obtained with each brood of sperm from nondysgenic and dysgenic males to their respective unirradiated controls. These values were 32.3%, 30.5% and 64.7% for brood 1, 2 and 3 respectively from nondysgenic males, and 14.1%, 56.1% and 71.4% for the same broods from dysgenic males. Thus the differential effect of X-rays in sperm broods 1, 2 and 3 was -18.2, +25.6 and +6.7% respectively. These results suggest that the synergistic effect may be due to the common component of X-ray and P element-induced genetic damage, namely chromosome breaks, and that the interaction of these lesions resulted in a greater than additive number of of unrestituted chromosome breaks and nonviable chromosomal rearrangements. PMID- 3039364 TI - Sexually transmitted diseases. PMID- 3039365 TI - Bilateral anterior interosseous nerve syndromes associated with cytomegalovirus infection. AB - The occurrence of bilateral brachial mononeuropathies presenting as anterior interosseous nerve syndromes in association with cytomegalovirus mononucleosis in an otherwise healthy adult is reported, a relationship not previously described. PMID- 3039366 TI - Experience with itraconazole in deep mycoses in northern Italy. PMID- 3039367 TI - The assessment of neurotoxicity in children. Electrophysiological methods. PMID- 3039368 TI - Characterization of Naegleria species by restriction endonuclease digestion of whole-cell DNA. AB - Whole-cell DNA in Naegleria spp. and two related genera was examined by restriction endonuclease digestion and fractionation of the fragments by agarose gel electrophoresis. Visual inspection of ethidium bromide-stained gels shows differences in banding pattern between N. fowleri, N. lovaniensis, N. gruberi, N. jadini, N. australiensis, Didasculus thorntoni and Willaertia magna, and between the two subspecies of N. australiensis. Even between strains belonging to the same species differences could be observed. Significant differences were seen between strains of N. fowleri according to the continent of origin, and a hypothesis on the ancestry and the dispersal of N. fowleri was deduced from it. A N. fowleri strain isolated from one of the very few cured human infections showed the most distinct pattern within the species. The considerable variation detected with serological and biochemical techniques between strains of N. australiensis as well as between strains of N. gruberi, was confirmed in the analysis of their whole-cell DNA. With this technique the existence of N. jadini, D. thorntoni, W. magna and two Naegleria strains as separate systematic entities is substantiated. PMID- 3039369 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 35-1987. A 21-year-old woman with a hepatic mass after treatment for amenorrhea. PMID- 3039370 TI - Signal clipping by the rod output synapse. AB - The properties of synapses between retinal neurons make an essential contribution to early visual processing. Light produces a graded hyperpolarization in photoreceptors, up to 25 mV in amplitude, and it is conventionally assumed that all of this response range is available for coding visual information. We report here, however, that the rod output synapse rectifies strongly, so that only potential changes within 5 mV of the rod dark potential are transmitted effectively to postsynaptic horizontal cells. This finding is consistent with the voltage-dependence of the calcium current presumed to control neurotransmitter release from rods. It suggests functional roles for the strong electrical coupling of adjacent rods and the weak electrical coupling of adjacent rods and cones. The existence of photoreceptor coupling resolves the apparent paradox that rods have a 25 mV response range, while signals greater than 5 mV in amplitude are clipped during synaptic transmission. We predict that the strengths of rod rod and rod-cone coupling are quantitatively linked to the relationship between the rod response range and the synapse operating range. PMID- 3039371 TI - Ankyrin binding to (Na+ + K+)ATPase and implications for the organization of membrane domains in polarized cells. AB - The interaction between membrane proteins and cytoplasmic structural proteins is thought to be one mechanism for maintaining the spatial order of proteins within functional domains on the plasma membrane. Such interactions have been characterized extensively in the human erythrocyte, where a dense, cytoplasmic matrix of proteins comprised mainly of spectrin and actin, is attached through a linker protein, ankyrin, to the anion transporter (Band 3). In several nonerythroid cell types, including neurons, exocrine cells and polarized epithelial cells homologues of ankyrin and spectrin (fodrin) are localized in specific membrane domains. Although these results suggest a functional linkage between ankyrin and fodrin and integral membrane proteins in the maintenance of membrane domains in nonerythroid cells, there has been little direct evidence of specific molecular interactions. Using a direct biological and chemical approach, we show here that ankyrin binds to the ubiquitous (Na+ + K+)ATPase, which has an asymmetrical distribution in polarized cells. PMID- 3039372 TI - New human and simian HIV-related retroviruses possess functional transactivator (tat) gene. AB - New human retroviruses antigenically related to HIV and even more closely to STLV III have been recently isolated from individuals from some West African countries. One of these viruses, HTLV-IVP, was reportedly isolated from lymphocytes of a healthy female prostitute. Another isolate, LAV-2FG, was obtained from an AIDS patient and third, SBL-6669, from an individual with lymphadenopathy. Current epidemiological studies indicate that some of these virus isolates cause immune deficiency whereas others may not or may be less efficient at inducing immune deficiency. Similarly, STLV-III apparently does not cause immune deficiency in its natural host, African green monkey. A novel feature of HIV is the possession of a gene termed tat, which is implicated in its pathobiology. We report here that, like HIV, HTLV-IVP, LAV-2FG (HIV-2) and SBL 6669, as well as STLV-IIIAGM possess the putative tat gene, irrespective of their pathogenic potential in vivo. Interestingly, HTLV-IVP/LAV-2FG long terminal repeat (LTR) is equally well transactivated by the HTLV-IVP/LAV-2FG and HTLV-IIIB tat function, HTLV-IIIB LTR responds better to its own tat function. PMID- 3039373 TI - Localization of the gene for familial adenomatous polyposis on chromosome 5. AB - Colorectal cancer is the second most common cancer in the United Kingdom and other developed countries in the West. Although it is usually not familial, there is a rare dominantly inherited susceptibility to colon cancer, familial adenomatous polyposis (FAP; also often previously called familial polyposis coli). During adolescence affected individuals develop from a few hundred to over a thousand adenomatous polyps in their large bowel. These are sufficiently likely to give rise to adenocarcinomas to make prophylactic removal of the colon usual in diagnosed FAP individuals. Adenomas may occur elsewhere in the gastrointestinal tract and the condition is often associated with other extracolonic lesions, such as epidermoid cysts, jaw osteomata and fibrous desmoid tumours. Adenomata have been suggested to be precancerous states for most colorectal tumours. Knudson has suggested that the mutation for a dominantly inherited cancer susceptibility may be the first step in a recessive change in the tumour cells, and that the same gene may be involved in both familial and non familial cases of a given tumour. Following up a case report of an interstitial deletion of chromosome 5 in a mentally retarded individual with multiple developmental abnormalities and FAP, we have now shown that the FAP gene is on chromosome 5, most probably near bands 5q21-q22. PMID- 3039374 TI - Prostaglandins and the synergism between VIP and noradrenaline in the cerebral cortex. AB - We have previously shown that vasoactive intestinal peptide (VIP) and noradrenaline (NA) interact synergistically to increase cyclic AMP levels in mouse cerebral cortical slices. The pharmacological mechanism of this synergism is the potentiation by NA, through alpha 1 adrenergic receptors, of the stimulatory effect of VIP on cAMP formation. A similar interaction has been confirmed in guinea pig cerebral cortex and in discrete nuclei of the rat hypothalamus. Furthermore VIP and NA interact synergistically to depress the spontaneous activity of identified neurons in rat neocortex. At the cellular level, this synergistic interaction suggests that VIP- and NA-containing neuronal systems may converge, at least in part, on the same target cells to increase cAMP levels in the cerebral cortex. At the molecular level, the interaction may occur at various steps in signal transduction, between receptors, intramembrane transduction processes or intracellular effector mechanisms. Here we report that the alpha 1-adrenergic potentiation of the increases in cAMP elicited by VIP involves the formation of arachidonic acid metabolites and is mimicked by prostglandins F2 alpha and E2. PMID- 3039375 TI - Micromolar concentrations of Zn2+ antagonize NMDA and GABA responses of hippocampal neurons. AB - NMDA (N-methyl-D-aspartate) receptors serve as modulators of synaptic transmission in the mammalian central nervous system (CNS) with both short-term and long-lasting effects. Divalent cations are pivotal in determining this behaviour in that Mg2+ blocks the ion channel in a voltage-dependent manner, and Ca2+ permeates NMDA channels. Zn2+ could also modulate neuronal excitability because it is present at high concentrations in brain, especially the synaptic vesicles of mossy fibers in the hippocampus and is released with neuronal activity. Both proconvulsant and depressant actions of Zn2+ have been reported. We have found that zinc is a potent non-competitive antagonist of NMDA responses on cultured hippocampal neurons. Unlike Mg2+, the effect of Zn2+ is not voltage sensitive between -40 and +60 mV, suggesting that Zn2+ and Mg2+ act at distinct sites. In addition, we have found that Zn2+ antagonizes responses to the inhibitory transmitter GABA (gamma-aminobutyric acid). Our results provide evidence for an additional metal-binding site on the NMDA receptor channel, and suggest that Zn2+ may regulate both excitatory and inhibitory synaptic transmission in the hippocampus. PMID- 3039376 TI - Site-specific phosphorylation induces disassembly of vimentin filaments in vitro. AB - Intermediate filaments are a major component of the cytoskeleton of eukaryotic cells. Although there appear to be at least five distinct classes of these filaments, cells of mesenchymal origin and most cells in culture contain the intermediate filament composed of the subunit protein vimentin. Vimentin exists in a nonphosphorylated as well as in a phosphorylated form, and there is increased phosphorylation of this protein when the filament undergoes marked redistribution in various cells. The role of phosphorylation on assembly disassembly and organization of the vimentin filament has remained obscure. We report here a stable and purified system allowing biochemical examination of vimentin filament assembly and disassembly. Using this in vitro system, we carried out stoichiometrical phosphorylations, using purified protein kinases. We obtained evidence for site-specific, phosphorylation-dependent disassembly of the vimentin filament. PMID- 3039377 TI - Evidence for local stimulation of ACTH secretion by corticotropin-releasing factor in human placenta. AB - The hypothalamic-pituitary-adrenocortical axis is activated in pregnancy and parturition. Levels of immunoreactive corticotrophin releasing factor (irCRF), immunoreactive adrenocorticotropic hormone (irACTH) and cortisol concentrations in maternal plasma are elevated throughout gestation, increase further during labour and fall precipitously after parturition. The placenta contains biologically active CRF and ACTH and it has been suggested that the placenta produces these peptides during pregnancy. Here we show that irCRF is located in the cytotrophoblast cells of placenta collected at term. Using a monolayer primary culture of human placental cells we have found that CRF stimulates secretion of peptides containing the ACTH sequence in the placenta in a dose dependent manner, as it does in the pituitary. This effect is reversed by a CRF antagonist and is mimicked by dibutyryl cyclic AMP and forskolin. Glucocorticoids, which suppress the secretion of pituitary ACTH, were found to have no influence on release of irACTH by the placenta. Oxytocin and prostaglandins stimulate irACTH and irCRF secretion from cultured placental cells and the irACTH-releasing activity of two prostaglandins is partially reversed by a CRF antagonist. Thus CRF may be involved in the paracrine regulation of placental irACTH secretion. PMID- 3039378 TI - Activation of a transposable element in the germ line but not the soma of Caenorhabditis elegans. AB - The genetic activity of transposable elements is tightly controlled in many species. Transposons that are relatively quiescent under certain circumstances can excise or transpose at greatly increased rates under other circumstances. For example, 'genomic shock' can activate quiescent maize transposons, 'cytotype' and tissue-specific splicing regulate Drosophila P factors, copy number controls Tn5 transposition in bacteria, and developmental timing affects the production of transposon-like intracisternal A-particles in mouse embryos. The Caenorhabditis elegans transposable element Tc1 is subject to both strain-specific and tissue specific control. Multiple copies of Tc1 are present in the genome of all C. elegans strains collected from nature. However, these elements are genetically active in only certain isolates. For example, in C. elegans variety Bristol transposition and excision of Tc1 are undetectable, but in variety Bergerac transposition and excision are frequent. Moreover, in variety Bergerac, Tc1 is about 1,000-fold more active in somatic cells than in germ cells. We have investigated the genetic basis for the germ/soma regulation of Tc1 activity. We have isolated mutants that exhibit increased frequencies of Tc1 excision in the germ line. The frequencies of Tc1 excision in the soma are unaltered in these mutants. These mutants also exhibit high frequencies of Tc1 germ-line transposition, and this results in a mutator phenotype. Nearly all mutator induced mutations are caused by insertion of Tc1. PMID- 3039379 TI - [Molecular biology of gamete conjugation]. AB - Contact between gametes takes place on a molecular basis via receptors on the surface of the egg coat. The functional part of this receptor glycoprotein is limited to the O-glycosidic-linked carbohydrate side chains, and a polypeptide chain of it may induce an acrosomal reaction on the sperm head, thereby inducing penetration of the sperm into the egg cell. Another function of sperm receptors is to block polyspermy, probably due to modification by limited proteolysis of the receptor glycoprotein. Fertilization is likely to be inhibited by steric hindrance of the receptor due to antibodies specific for glycoproteins of the egg coat. The study of gamete interactions on a molecular basis may serve as a model for intercellular recognition in general. PMID- 3039380 TI - Changes in central serotoninergic transmission affect clonidine analgesia in monkeys. AB - The effects of changes in central serotoninergic transmission on clonidine analgesia were assessed in monkeys. The minimum electrical current required for producing jaw opening is referred to as the pain threshold. Pain was induced by electrical stimulation of tooth pulp afferents. In the first series of studies, intracerebroventricular administration of clonidine (5-30 micrograms) produced dose-dependent analgesia in monkeys. The clonidine-induced analgesia was abolished or attenuated by prior injection of the animals with p chlorophenylalanine or 5,7-dihydroxytryptamine into the third cerebral ventricle. On the other hand, pretreatment of the animals by injecting 5-HT or its precursor 5-hydroxytryptophan into the cerebral ventricle potentiated the clonidine-induced analgesia in monkeys. In the second series of experiments, administration of clonidine (1-10 micrograms) into the diencephalic periventricular gray (of the anterior hypothalamic portion), the periaqueductal gray, or the dorsal raphe nuclei also produced dose-dependent analgesia in monkeys. The analgesia induced by clonidine injection into the diencephalic periventricular gray or the periaqueductal gray was effectively antagonized by pretreatment of the animals by injecting two 5-HT receptor antagonists (such as ketanserine and methysergide) into the diencephalic periventricular gray or the periaqueductal gray. The clonidine-induced analgesia in monkeys was not affected by pretreatment of the animals with injections of either ketanserine or methysergide into the dorsal raphe nuclei. The results suggest that the functional activity of central 5-HT neurons correlate well with the analgesic sensitivity of clonidine microinjected centrally. In addition, the analgesia induced by clonidine microinjected into the diencephalic periventricular gray or the periaqueductal gray was mediated by the 5-HT receptors at the site of injection. PMID- 3039381 TI - Blockade of gamma-aminobutyric acid-receptors of the B-subtype inhibits the dopamine-induced enhancement of the release of cholecystokinin-like immunoreactivity from slices of rat dorsal caudatoputamen. AB - When slices of rat dorsal caudatoputamen (= neostriatum) are incubated in vitro, cholecystokinin-like immunoreactivity (CCK-LI) is released upon addition of veratridine (3.75 mumol/l). This release is affected by dopamine and by gamma aminobutyric acid (GABA)-receptor agonists. Dopamine enhances the release by stimulating dopamine D2-receptors and decreases it via D1-receptors. GABAA receptor agonists enhance the veratridine-induced release of CCK-LI, while GABAB receptor agonists decrease it. In the present investigation, it was examined whether GABA-receptors are involved in the effect which dopamine exerts via D2 receptors. The GABAA-receptor antagonist bicuculline (10 mumol/l) and the blocker of the GABAA-receptor ionophore picrotoxin (1 mumol/l) did not affect the dopamine (0.1 mumol/l)-induced increase in the release of CCK-LI. However, the GABAA-receptor agonist muscimol (1 mumol/l) not only enhanced the release of CCK LI, but also prevented a further enhancement by dopamine (0.1 mumol/l). This effect of muscimol was blocked by bicuculline (10 mumol/l). In the presence of delta-amino-n-valeric acid (0.1 mmol/l), which has been described to block GABAB receptors, dopamine no longer enhanced the veratridine-induced release of CCK-LI. delta-Amino-n-valeric acid also inhibited the pronounced enhancement of the release of CCK-LI caused by dopamine (0.1 mumol/l and 1 mumol/l in the presence of the preferential D1-receptor antagonist SCH 23390. The effect of delta-amino-n valeric acid persisted in the presence of bicuculline (10 mumol/l and 100 mumol/l).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3039382 TI - Metabolism of bradykinin and angiotensin I by human basilar artery and rabbit aorta. AB - The role of angiotensin converting enzyme in the metabolism of bradykinin and angiotensin I by in vitro human basilar artery and rabbit aorta was studied. On both human basilar artery and rabbit aorta concentration-effects curves to angiotensin I were significantly attenuated by captopril at a concentration which had no effect on bradykinin responses on both tissues. The metabolism of bradykinin and angiotensin I was studied using high performance liquid chromatography. Both peptides were broken down by human basilar artery and rabbit aorta in a similar fashion. The breakdown of angiotensin I but not bradykinin was significantly attenuated by captopril. 1,10-phenanthroline did attenuate breakdown of bradykinin but this was found not to be significant compared with controls. The results confirm that angiotensin converting enzyme is present in both these tissues and is important for the conversion of angiotensin I to angiotensin II. It appears that other peptidases are important in the breakdown of kinins by these tissues and should be taken into account when investigating the mechanism of action of such peptides on these vascular preparations. PMID- 3039383 TI - [Facial paralysis]. PMID- 3039384 TI - [Constipation in the elderly]. PMID- 3039385 TI - [Regulation of the efficacy of adrenergic synaptic transmission by autoadrenoreceptors in rat brain slices]. AB - A hypothesis of the role of autoadrenoreceptors in the processes of synaptic depression, facilitation, PSP amplitude stabilization and amplitude increase with the stimulation frequency growth is described by means of a mathematical model. Synaptic depression is realized by activation of alpha-autoadrenoreceptors, synaptic facilitation--by beta-autoadrenoreceptors activation. A "stable secretion zone" is formed between the autoadrenoreceptor activation curves at which noradrenaline secretion is stabilized. Experimental studies of H3-NA secretion from rats cerebral cortex slices showed that autoadrenoreceptors really form a stable secretion zone. The presence of stable secretion zone provides for self-facilitation and stabilization of H3-NA secretion elicited by 5-fold consecutive K+-depolarizations of slices with time interval 30 min. The absence of the stable secretion zone leads to destabilization of H3-NA secretion. With an increase of the stimulation intensity the curves of autoadrenoreceptors activity and stable secretion zone shift toward high noradrenaline concentrations. PMID- 3039386 TI - [Cholinergic modulation of synaptic transmission in the spinal cord of the frog]. AB - Superfusion of the isolated frog spinal cord by the Ringer solution containing arecoline (10 mumol/l) evoked depolarization and increase of the input resistance and PSP amplitude of motoneurons. Depolarizing electrotonic potentials and reflex discharges in the ventral roots also increased, but duration of dorsal root potentials decreased. The observed arecoline facilitation of synaptic transmission in the spinal cord has postsynaptic nature evoked by motoneuron M2 cholinoreceptor activation and bound to an increase of the cyclic nucleotide metabolism. Arecoline inhibited the synaptic transmission in the spinal cord under conditions when its postsynaptic action was eliminated. This effect was due to presynaptic M1-cholinoreceptor activation without changing the cyclic nucleotide metabolism. PMID- 3039387 TI - Chemo- and immunotherapy of an adenovirus-induced transplantable sarcoma in hamsters. AB - The subcutaneous transplantable adenovirus sarcoma (TAVS) in hamsters was created in 1972 at the Oncological Research Institute and maintained by serial subcutaneous transplantation. It showed the highest sensitivity against some alkylating drugs and antimetabolites--cyclophosphamide, sarcolysine, methotrexate and 6-mercaptopurine. In some degree, TAVS is able to differentiate antitumor drugs of one group by the intensity of their antitumor activity. Among the drugs, cyclophosphamide was the most active and 6-mercaptopurine and 5-fluorouracil the least active. Antitumor drugs of plant origin, antibiotics and methyl-CCNU had a weak activity on subcutaneous TAVS. Intramuscular TAVS showed a similar sensitivity, with some differences for olivomycin, bruneomycin and vinblastine. Intracerebral TAVS was sensitive against antitumor agents penetrating through the hemato-encephalic barrier and which were active against its subcutaneous and intramuscular form. BCG vaccine and levamisole applied separately did not show any activity on the growth of TAVS. Combined immunochemotherapy with cyclophosphamide plus BCG gave a better enhancement of the antitumor effect of the cytostatic than that of the combination of methotrexate plus BCG and cyclophosphamide plus levamisole. These results showed that TAVS in hamsters may be used as a suitable experimental model for pharmacological studies of antitumor agents and combined immunochemotherapy. PMID- 3039388 TI - [Local treatment of obstructive uric acid calculi]. AB - Dissolution of uric acid calculi could be obtained by oral or parenteral urinary alcalinization, but this method cannot apply to the case of obstructive calculi. Nineteen obstructive calculi in 18 patients were treated by in situ alcalinization through a percutaneous nephrostomy catheter (PCN). Eight patients were initially anuric, 7 of whom from an obstructed solitary kidney and 1 from a bilateral obstructive lithiasis. Fifteen calculi were located in the ureter, 3 in the uretero-pelvic junction and 1 in the pelvis. After 48 h of urinary diversion through PCN, an isotonic sodium bicarbonate solution (14 g %) was continuously infused at an average flow rate of 2.8 l/24 h, through either an unique PCN, or a 2 PCN-irrigation circuit in the 7 cases with permanently obstructive calculus. Fifteen calculi (80%) were completely dissolved after 3 to 13 days of alcalinization (average 5.8 days). One large calculus was reduced by 3/4 and further removed by percutaneous lithotripsy. Three patients underwent ureterotomy after 9 to 11 days of uneffective treatment. Local alcalinization is an effective and non invasive treatment for obstructive uric acid calculi, and is logically associated with the necessary urinary diversion. PMID- 3039389 TI - Detailed analysis of heterogeneity of [3H]naloxone binding sites in rat brain synaptosomes. AB - The experiments reported in this paper address the question of heterogeneity of [3H]naloxone binding sites in rat brainstem synaptosomal preparations at 23 degrees C in the presence of 100 mM sodium chloride. Kinetic analysis in the presence of 0.4, 4 and 10 nM [3H]naloxone gave pseudo-first order association rate of 0.9 +/- 0.04, 1.23 +/- 0.08 and 1.06 +/- 0.08 min-1, respectively. The dissociation of a 1 nM [3H]naloxone receptor complex was biphasic with dissociation rate constants of 1.8 and 0.4 min-1. On the other hand, dissociation of 10 nM [3H]naloxone was monophasic with a kd of 1.1 min-1. Two subpopulations of binding sites were also observed by steady state binding studies, with Kd values of 0.5 and 3.4 nM and a ratio of high to low affinity sites of 1:9. Heterogeneity of [3H]naloxone binding sites could be seen by displacement studies performed with opioid peptides and alkaloids. We suggest that our data best fits a model with two independent naloxone binding sites wherein either one or both undergo a multi-step interaction with ligand. PMID- 3039390 TI - Binding of [3H]muscimol to calf cerebrocortical synaptic membranes and the effects of sulphur-containing convulsant and non-convulsant compounds. AB - Endogenous and xenobiotic sulphur-containing convulsant and non-convulsant compounds containing structural moieties of, or bearing a structural resemblance to, GABA and homocysteine were tested in binding studies for their potency in displacing the GABA-mimetic [3H]muscimol from specific, high-affinity sites (Kd = 3.6 nM; Bmax = 3.94 pmol/mg protein) on freeze-thawed, Triton-treated calf-brain synaptic membranes. The xenobiotic convulsants, 4-mercaptobutyric acid (MBA), 3 mercaptopropionic acid (3-MPA) and 2-mercaptopropionic acid (2-MPA) were found to be two-site competitive inhibitors exhibiting apparent inhibition affinity constants (Kiapp) of 5000 microM, 3750 microM, and 4800 microM, respectively; while homocysteic acid (Kiapp = 4800 microM) was shown to be a one-site partial competitive inhibitor. Intermediary metabolites of methionine: S-adenosyl-L homocysteine, L-cysteine, the convulsant L-homocysteine, and its non-convulsant disulphide oxidation product, homocystine, were found to be one-site partial competitive inhibitors exhibiting Kiapp values of 5750 microM, 8350 microM, 5000 microM, and 510 microM, respectively. The endogenous anticonvulsant neuroeffector, taurine, and the tripeptide, reduced glutathione (GSH) were shown to be, respectively, one-site (Ki = 20 microM) and two-site (Kiapp = 4300 microM) competitive inhibitors of [3H]muscimol binding. These findings are discussed with regard to a previously proposed mechanism for the convulsant action of homocysteine. PMID- 3039391 TI - Na+-K+-ATPase activity of glial, neuronal, and synaptosomal enriched fractions from normal and freezing-injured rabbit cerebral cortex. AB - This paper investigates the kinetic parameters of Na+-K+-ATPase in glial, neuronal, and synaptosomal enriched fractions isolated from rabbit cerebral cortex. Under normal conditions, kinetic parameters-Vmax and KK+0.5- of Na+-K+ ATPase are the same in the three fractions, suggesting that this enzyme behaves as the same molecular entity. Following a cryogenic lesion, the alterations of these parameters appear to be different in the different fractions. These data suggest that the same enzyme exhibits various responses when exposed to the same pathological event. The dissimilar lipid composition of the Na+-K+-ATPase environment, and/or different adaptative responses to abnormal ion concentrations in glial, neuronal, and synaptosomal fractions could account for these different responses. PMID- 3039392 TI - Selective infections of olfactory and respiratory epithelium by vesicular stomatitis and Sendai viruses. AB - Following intranasal instillation of vesicular stomatitis virus (VSV) in mice there was an extensive infection of the olfactory epithelium in contrast to a minimal involvement of the respiratory epithelium. Sendai virus (SV), on the other hand, caused an extensive infection of the respiratory epithelium and only minimal infection of the olfactory mucous membrane. VSV budded from basolateral surfaces of supporting cells and olfactory neurons, but not from their apical surfaces or the ciliated bulbous endings of the olfactory neuron dendrites. This asymmetric release of VSV favoured neuroinvasion. The virus spread along the olfactory nerves to the glomeruli in the olfactory bulbs after which it propagated transneuronally into the rest of the brain. SV budded only from the apical surface of respiratory epithelial cells, was released into the air passages, and there were no signs of invasion into the olfactory bulbs. Inoculation of the olfactory mucous membrane is a useful procedure for studies on selectivity of attack on peripheral neurons by viruses and on mechanisms of virus invasion of the nervous system in vivo. PMID- 3039393 TI - Effect of passive immunization against corticotropin-releasing factor (CRF) on the postadrenalectomy changes of CRF binding sites in the rat anterior pituitary gland. AB - The concentration of corticotropin-releasing factor (CRF)-binding sites decreases in the rat anterior pituitary after adrenalectomy; this change may be related either to a direct effect of the circulating glucocorticoids at the pituitary level or to a desensitization of CRF receptors through an increased CRG release in hypophysial portal blood. In order to examine the latter possibility we have measured plasma adrenocorticotropin hormone (ACTH) levels and the number of anterior pituitary CRF binding sites in sham-operated and 24-hour adrenalectomized rats after blockade of endogenous CRF by passive immunization with an antiserum anti-rat CRF (CRF-AS), or after injection of normal rabbit serum (NRS). In NRS-injected rats, after sham operation, plasma ACTH concentration increased (227 +/- 34 vs. 118 +/- 19 pg/ml in controls) without change in CRF-binding sites capacity (20.7 +/- 2.6 vs. 24.6 +/- 3.5 fmol/mg protein in controls). Adrenalectomy induced a large rise in plasma ACTH (785 +/- 89 pg/ml) and a decrease in the number of CRF-binding sites (12.2 +/- 1.7 fmol/mg protein). After CRF-AS injection, plasma ACTH was normalized in sham-operated animals (149 +/- 24 pg/ml) and significantly reduced in adrenalectomized rats (472 +/- 76 pg/ml); the adrenalectomy-induced decrease in the number of CRF binding sites was unaffected by the CRF-AS administration (12.2 +/- 1.7 fmol/mg protein). The administration of dexamethasone to adrenalectomized rats significantly reduced plasma ACTH concentrations (23.3 +/- 10.6 pg/ml) and prevented the loss in CRF-binding sites capacity (20.7 +/- 1.3 fmol/mg protein).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3039394 TI - Stimulation of luteinizing hormone release and cyclic nucleotide production by arachidonic acid in cultured pituitary gonadotrophs. AB - Gonadotropin-releasing hormone (GnRH) stimulates luteinizing hormone (LH) release and cyclic guanosine 3',5-cyclic monophosphate (cGMP) production in rat anterior pituitary cells through a calcium-dependent activation mechanism that involves increased phospholipid turnover and liberation of arachidonic acid. In enriched pituitary gonadotrophs, LH release was stimulated by arachidonic acid and its oxygenated metabolite, 5-hydroxy-6,8,11,14-eicosatetraenoic acid (5-HETE), in a dose-dependent manner. The prominent LH responses of purified gonadotrophs to arachidonic acid suggest that the secretory actions of arachidonate are exerted primarily on the gonadotroph and do not involve the participation of other pituitary cell types. Preincubation of pituitary cells with stimulatory concentrations of arachidonic acid for up to 120 min did not alter the subsequent LH responses elicited by GnRH, indicating that the secretory mechanism was unimpaired by arachidonate treatment and that no cross-desensitization occurs during sequential exposure of gonadotrophs to the two stimuli of LH release. Cyclic adenosine 3',5-monophosphate (cAMP) production was stimulated by 10 microM arachidonic acid to the same degree (about 2-fold) as by GnRH, but did not parallel the progressive LH response to higher arachidonate concentrations. cGMP production was initially stimulated by addition of arachidonic acid but returned to the control value after 5 min, whereas GnRH typically elicited a prolonged cGMP response. In contrast to the calcium-independent action of arachidonic acid, the stimulatory effect of 5-HETE on LH release required the presence of extracellular Ca2+, as previously observed for GnRH. These findings demonstrate that arachidonic acid and its metabolite, 5-HETE, partially reproduce the actions of GnRH upon LH release and cyclic nucleotide production.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3039395 TI - Distribution of neurotransmitter binding sites in the cat median eminence. AB - Receptor autoradiography was used to determine the distribution of binding sites for monoaminergic and cholinergic neurotransmitters in the cat median eminence. Results showed that binding sites were differentially distributed in that [3H]-p aminoclonidine (an alpha-2-adrenergic ligand) and [3H]pyrilamine (an H1 histaminergic ligand) binding were concentrated in the medial region of the external layer of the median eminence while [3H]lysergic acid diethylamide (a serotonergic ligand) and [3H] quinuclidinyl benzilate (a muscarinic cholinergic ligand) binding were concentrated within the lateral region of the external layer. These patterns are coincident with the differential distribution of hypophysiotropic hormones in the median eminence, suggesting that monoamines and acetylcholine may regulate the release of these hormones at the level of the median eminence. PMID- 3039396 TI - Irreversible labelling of delta-opioid receptors in rat brain and neuroblastoma cells by [3H]azido-DTLET: characterization of subunits and autoradiographic visualization of the covalent binding. AB - [3H]Az-DTLET (Tyr-D-Thr-Gly-Phe(pN3)-Leu-Thr), a photoaffinity probe for delta opioid receptors binds to a single class of sites in rat brain membranes with a high affinity (KD = 1.66 nM). The selectivity index of Az-DTLET (KI delta/KI mu = 0.036) is better than that of its precursor DTLET (0.053). Rat brain or neuroblastoma glioma cells membranes were incubated with 10 nM [3H]Az-DTLET, washed and irradiated with U.V. After irradiation a fraction (20-30%) of specific binding was found to remain indissociable after 10 min at 60 degrees C and was considered as irreversible. This fraction increased as a function of the irradiation time. The radioactivity irreversibly bound to rat brain membranes, solubilized by sodium cholate, was associated with high molecular weight species (200,000 daltons). In denaturing conditions (SDS 2%), the [3H]Az-DTLET specific binding was associated with molecular components of 45-50 K and 90-100 K daltons. In contrast, when opioid receptors were prelabelled by [3H]Az-DTLET, solubilized by Na-cholate and irradiated, the radioactivity was only recovered with subunits of 45-50 K daltons. The autoradiographic localization of the irreversibly bound [3H]Az-DTLET in rat brain was identical to that of reversibly bound [3H]DTLET or [3H]Az-DTLET. These results suggest that [3H]Az-DTLET represents an adequate specific probe for studies on the structure, function and anatomical distribution at light and even electron microscopic level of delta-opioid receptors. PMID- 3039397 TI - Superior sensitivity of computed tomographic scanning to magnetic resonance imaging in metastatic neoplasia: two case reports. AB - A case of carcinoma of the breast and a case of melanoma metastatic to the brain were evaluated by both magnetic resonance imaging (MRI) and enhanced computed tomography (CT). In these two cases, enhanced CT was more sensitive and more clearly delineated the extent of disease. Does diagnostic imaging of central nervous system metastatic disease require both MRI and enhanced CT? PMID- 3039398 TI - Supratentorial gliomas: surgical considerations and immediate postoperative results. Gross total resection versus partial resection. AB - Forty-two patients with supratentorial gliomas not involving the basal ganglia (extraganglionic) were studied pre- and postoperatively with computed tomographic (CT) scans to evaluate the effect of the extent of surgical resection on the immediate postoperative results. Thirty-three patients (79%) had malignant astrocytic gliomas (glioblastoma or anaplastic astrocytoma), 4 patients (10%) had well-differentiated astrocytomas, and 5 (12%) had oligodendrogliomas. The median age was 58 years, and the median Karnofsky rating was 70. There was no operative mortality. Six patients (14%) had surgical complications. A gross total resection was defined as the absence of any abnormal enhancement on the postoperative CT scan. A nearly gross total resection had been accomplished when less than 10% of the preoperatively enhancing mass was still seen. A partial resection was indicated by the presence of more than 10% of the enhancing lesion on the postoperative CT scan. A gross total or nearly gross total resection was accomplished in 36 patients (86%), and an improved or stable postoperative neurological status was present in 35 of these patients (97%). In contrast, the rate of neurological morbidity after a partial resection was 40%. Supratentorial extraganglionic gliomas, regardless of their histological type, generally were well-circumscribed lesions except at the level of the ventricular wall, where glioblastomas and anaplastic astrocytomas blended with the subependymal white matter from which they seemed to arise. PMID- 3039399 TI - Intracranial catecholamine secreting paragangliomas. AB - Three intracranial catecholamine-secreting paragangliomas are described. They involved a glomus jugulare, a sphenopalatine ganglion and the clivus and upper cervical spine respectively. The extent of the tumours was shown by CT and MRI. They were all highly vascular with a substantial blood supply from systemic arteries which was subjected to particulate embolisation, followed in two cases by surgery. The importance of studying any tumour which could possibly be a paraganglioma for hormonal and, especially, for catecholamine secretion prior to any invasive procedure, including angiography and embolisation, is emphasised: all such procedures should be covered with catecholamine blocking agents and, in addition, carefully monitored with resuscitation facilities immediately available. PMID- 3039400 TI - Intrathecal 6-hydroxydopamine or cervical spinal hemisection reduces norepinephrine content, but not the density of alpha 2-adrenoceptors, in the cat lumbar spinal enlargement. AB - The effect of the intrathecal administration of the catecholaminergic neurotoxin 6-hydroxydopamine, or of hemisection of the spinal cord at the Cl level, on the density of alpha 2-adrenoceptors and on the norepinephrine, dopamine, and serotonin content in the cat lumbar spinal enlargement was determined 2, 7 or 21 days after performance of each type of lesion. The intrathecal administration of 6-hydroxydopamine produced a time-dependent reduction of norepinephrine content in the cat lumbar spinal enlargement (95% reduction at 21 days) without significantly altering the serotonin content in this same tissue of the same cats. The dopamine content of the dorsal horn was not changed significantly, whereas ventral horn dopamine content was depleted after intrathecal 6 hydroxydopamine. alpha 2-Adrenoceptor binding site density was not significantly different from control either 2 or 21 days after 6-hydroxydopamine, but was increased significantly (50%) over the control density 7 days after 6 hydroxydopamine. Hemisection of the cervical spinal cord produced a bilateral 40 60% reduction of norepinephrine content in both the dorsal and ventral horns of the cat lumbar spinal enlargement 7 and 21 days later. Cervical hemisection did not significantly alter the alpha 2-adrenoceptor binding site density in these same cats either 2, 7, or 21 days after performance of the lesion. It is concluded that alpha 2-adrenoceptors located on the terminals of descending noradrenergic or other spinopetal fibers do not represent a significant fraction of the total population of alpha 2-adrenoceptors present in the dorsal or ventral cat lumbar enlargement. PMID- 3039401 TI - Angiotensin converting enzyme immunohistochemistry in rat brain and pituitary gland: correlation of isozyme type with cellular localization. AB - We have localized angiotensin converting enzyme in rat brain and pituitary gland immunohistochemically with an anti-rat lung angiotensin converting enzyme monoclonal antibody. The distribution of immunoreactive angiotensin converting enzyme is identical with that of binding sites for the angiotensin converting enzyme inhibitor, [3H]captopril. Most intense staining is in the choroid plexus and subfornical organ, with intermediate values in the caudate-putamen, globus pallidus, entopeduncular nucleus, pars reticulata of the substantia nigra, posterior pituitary and anterior pituitary. Lower levels are observed in the supraoptic and paraventricular nuclei of the hypothalamus. Within the basal ganglia angiotensin converting enzyme immunoreactivity is distributed throughout the neuropil; no cell bodies are stained, even after colchicine treatment. The punctate pattern of immunoreactivity in the anterior pituitary corresponds to the distribution of endothelial cells. The posterior pituitary is stained diffusely. Angiotensin converting enzyme is increased by 45% in the posterior lobe after pituitary stalk section, demonstrating that this diffuse staining is associated with pituicytes. Antibody specificity was demonstrated by the immunoaffinity purification of angiotensin converting enzyme to homogeneity from crude tissue extracts using anti-angiotensin converting enzyme antibody and protein A sepharose. The apparent molecular weight by sodium dodecyl sulfate polyacrylamide gel electrophoresis of lung, choroid plexus and anterior pituitary angiotensin converting enzyme is 175,000. In the substantia nigra and caudate putamen, where angiotensin converting enzyme is localized to neuronal as opposed to epithelial cells, the molecular weight is 165,000. The pituicyte angiotensin converting enzyme of the posterior pituitary is 170,000 daltons. PMID- 3039402 TI - The initiation and spread of epileptiform bursts in the in vitro hippocampal slice. AB - We recorded spontaneous synchronized epileptiform bursts from hippocampal slices from guinea pig using an array of 16 extracellular electrodes placed over the stratum pyramidale of CA2 and CA3. The slices were made epileptogenic with the GABA antagonist picrotoxin (or occasionally penicillin). We found that spontaneous bursts always originate at a discrete focus at or near CA2. These bursts spread smoothly and uniformly across CA3 at an average velocity of 0.13 m/s. This velocity is slower than the conduction velocity of the Schaffer collaterals or mossy fibers. Picrotoxin produced afterdischarges following the initial primary burst, and these afterdischarges were found to originate and spread in a fashion nearly identical to the primary burst. These results indicate that CA2 is a unique region which must possess unusual cellular and/or synaptic connectivity properties which result in a decreased threshold for initiation of epileptiform activity. We consider several hypothetical patterns of local synaptic connectivity in the light of these results, and we discuss the possible role of residual inhibition in limiting the spread of synchronized discharges. PMID- 3039403 TI - Models of the cellular mechanism underlying propagation of epileptiform activity in the CA2-CA3 region of the hippocampal slice. AB - We have shown experimentally in the previous paper that spontaneous epileptiform activity, as recorded by extracellular field potentials, propagates smoothly across the CA2-CA3 region of the convulsant-treated hippocampal slice of the guinea pig at velocities of about 0.1 m/s. In the present paper, we used computer simulations of either 500 or 1000 cell arrays of model neurons to examine possible mechanisms underlying this propagation. We show that propagation of epileptiform field potentials can be explained plausibly by slow conduction along axons interconnecting CA2-CA3 neurons, provided that there are sufficiently many interconnections. This propagation can take place even if the interconnections occur randomly. The number of interconnections required decreases as the number of synchronously activated cells initiating a population burst increases. Axonal propagation at 0.1 m/s appears to be a plausible assumption, since conduction velocities along Schaffer collaterals have been experimentally estimated to be as slow as 0.2 m/s, and small recurrent collaterals are likely to conduct more slowly than the main axonal branches. If spontaneous synchronized population bursts are initiated by activity in four or fewer cells, then our model requires, for smooth field potential propagation, more interconnections than are believed to occur on the basis of dual intracellular recording. PMID- 3039404 TI - The nigrotectal projection in the cat: an electron microscope autoradiographic study. AB - Recent evidence indicates that the nigrotectal tract plays an important role in regulating the premotor responses of cells in the in the intermediate gray layer of the superior colliculus. The purpose of the present study was to characterize the ultrastructure of nigrotectal terminals and of their postsynaptic targets in the intermediate gray layer. Nigrotectal terminals were identified in the electron microscope by labeling them autoradiographically, following injections of tritiated proline into the substantia nigra pars reticulata. The majority of nigrotectal terminals contain a high proportion of pleomorphic vesicles and form symmetrical synaptic contacts. Most of these terminals synapse with small dendritic profiles (2.00 micron +/- 0.83 SD), which may be the distal dendrites of neurons in the intermediate gray layer. Less than 10% of the labeled contacts are made with cell bodies or initial axonal segments. PMID- 3039405 TI - Naloxone-induced depolarization and synaptic activation of myenteric neurons in morphine-dependent guinea pig ileum. AB - To investigate the cellular basis of opiate dependence, intracellular microelectrodes were used to record from both electrophysiologically defined classes of neurons (S and AH) in myenteric plexus longitudinal muscle preparations from morphine pretreated guinea pigs. These preparations responded to naloxone with the characteristic contraction of the longitudinal smooth muscle, indicative of morphine dependence. Depolarization in response to naloxone was observed in 42% of S neurons, but there were no consistent changes in input resistance. In some cells the depolarization was reduced or abolished after blockade of synaptic transmission, suggesting that it was due in part to the release of an excitatory transmitter producing a slow depolarization in the impaled neuron. Synaptic activation of S neurons during withdrawal was further indicated by the observation that fast postsynaptic potentials appeared after abrupt displacement of morphine from its receptors by naloxone. Morphine withdrawal, therefore, involves both the final motor neurons and interneurons. During naloxone-induced withdrawal, 25% of S neurons discharged action potentials. In contrast, no action potentials were discharged in AH neurons. Furthermore, naloxone did not alter the resting membrane potential, input resistance, soma action potential configuration, or slow hyperpolarization following a soma spike in AH neurons. The specificity of the withdrawal response for S neurons and the relatively small proportion of neurons involved suggests that morphine withdrawal occurs in quite specific neuronal circuits in the myenteric plexus. PMID- 3039406 TI - Incorporation of vesicular antigens into the presynaptic membrane during exocytosis at the frog neuromuscular junction: a light and electron microscopy immunochemical study. AB - We have studied the incorporation of vesicular membrane antigens into the presynaptic membrane during exocytosis of neurotransmitter at the frog neuromuscular junction. In a preliminary series of experiments, we first confirmed by electron microscopy that the synaptic vesicles are labelled following incubation with rabbit antisynaptic vesicle antibody of neuromuscular junction cross sections (cytoplasm and organelles reached by the antibodies). In a second series of experiments, intact neuromuscular junctions were stimulated with black widow spider venom and fixed with paraformaldehyde. The presence or absence of vesicular antigens in the presynaptic membrane was monitored with rabbit antisynaptic vesicle antibody and revealed with a second antibody coupled to peroxidase. In light microscopy, the labelled neuromuscular junctions are almost completely restricted to muscles stimulated with black widow spider venom and incubated with rabbit antisynaptic vesicle antibody. Only a few control muscles (not stimulated with black widow spider venom, but incubated with rabbit antisynaptic vesicle antibody) had labelled neuromuscular junctions. All control neuromuscular junctions, not incubated with rabbit antisynaptic vesicle antibody were unlabelled. Electron microscopy indicated that it is the presynaptic membrane of intact stimulated neuromuscular junctions which is labelled. In these intact neuromuscular junctions, the synaptic vesicles are usually unlabelled. Electron microscopy also indicated that the presynaptic membrane of only one type of control junctions (not stimulated with black widow spider venom, but incubated with rabbit antisynaptic vesicle antibody) is rarely and weakly labelled. Other types of controls (not incubated with rabbit antisynaptic vesicle antibody) are never labelled. Therefore our results are consistent with the incorporation of vesicular antigens into the plasma membrane during exocytosis produced by the black widow spider venom. The low level of labelling of unstimulated neuromuscular junctions suggest a rather complete retrieval of the vesicular proteins during endocytosis. PMID- 3039407 TI - Combined central and peripheral myelinopathy. AB - We studied clinical, electrophysiologic, and nerve biopsy findings in two men with evidence of severe central and peripheral demyelinating disease. These patients may be rare examples of MS with associated severe, chronic, clinically evident peripheral demyelinating neuropathy. Alternatively, they may be cases of some other form of combined central-peripheral myelinopathy or fortuitous coincidence of MS with idiopathic inflammatory-demyelinating neuropathy. There have been only five other reports of clinically evident combined central peripheral myelinopathy, but there have also been reports of only electrophysiologic or nerve biopsy evidence (without clinical manifestation) of peripheral neuropathy in MS patients. PMID- 3039408 TI - Brachial neuropathy after brachial artery-antecubital vein shunts for chronic hemodialysis. AB - Peripheral mononeuropathies may complicate distal arteriovenous fistulas for chronic renal dialysis. We observed three diabetic patients who developed pain, paresthesias, and weakness in the distribution of the median, ulnar, and radial nerves shortly after construction of proximal brachial artery-antecubital vein fistulas. EMG confirmed multiple distal nerve injuries. All three patients improved after shunt banding or ligation. Twenty additional patients with proximal shunts were examined for risk factors for brachial neuropathy. Although all patients had severe atherosclerosis and many had polyneuropathy, we identified no predictive risk factors other than diabetes. PMID- 3039409 TI - Peripheral neuropathy associated with Crohn's disease. AB - Peripheral neuropathy in Crohn's disease has been described, to date, only with vitamin B12 deficiency or as due to oral metronidazole treatment. We report the association of Crohn's disease and peripheral neuropathy in two patients in whom neither of these pathogenetic factors of nerve damage apply. The CSF of both was normal. Patient 1 has had Crohn's disease for 12 years with predominantly sensory distal neuropathy and recurrent course related to worsening and improvement of the enteritis. Patient 2 had Crohn's disease some years before symptoms of recurrent sensory loss in the feet. Axonal degeneration was the mechanism of nerve damage in both patients. PMID- 3039410 TI - AIDS. Case for diagnosis series, 1987. PMID- 3039411 TI - [Malignant fibrous histiocytoma of the greater pectoral muscle. Observations and echotomographic study of a case]. PMID- 3039412 TI - Cylindroma of the scalp. AB - A patient is presented with a large dermal cylindroma of the whole scalp. The origin of the tumor from sweat glands has been well accepted, but it is still not clear whether the tumor arises from the eccrine or the apocrine glands. Differential diagnosis and surgical treatment are discussed together with the typical clinical and pathological features of this tumor. PMID- 3039413 TI - Benign pedunculated tumor of the esophagus. AB - Benign pedunculated tumors of the esophagus are rare, even large tumors often remain asymptomatic. Recent experiences with two patients are described, who only complained of recurrent appearance of the tumor in their mouth. In barium studies the esophageal dilatation can incorrectly suggest achalasia or cardiospasm, whereas the tumor itself is overlooked. Even at endoscopy the tumor can be missed, as it is covered with normal epithelium. Histology of a biopsy often shows no abnormalities. Once the diagnosis is made, resection is strictly indicated, especially to prevent acute laryngeal obstruction. In one patient surgical removal was performed via cervical esophagotomy, in the second by endoscopy. PMID- 3039414 TI - [Evaluation of the hemorheologic effects of the eicosapentaenoic and docosahexaenoic acids in asymptomatic dyslipidemic and arteriopathic subjects]. AB - The low incidence of cardiovascular disease in Greenlandic Eskimos is related to the effect of dietary omega-3 polyunsaturated fatty acids. An important anti atherosclerotic mechanism of compounds such as EPA and DHA consist in the modulation of cell membrane and properties. A change in the lipid composition of the diet and a dietary supplementation with EPA and DHA contribute to suggested potential therapeutic effect on peripheral vascular disease. The Authors report the results obtained by Doppler C.W. ultrasound in hyperlipidemic-arteriopathic patients. PMID- 3039415 TI - Coats' disease in a renal transplant recipient. AB - We report the first case, to our knowledge, of Coats' disease, an idiopathic exudative retinopathy, in a renal transplant recipient. Due to certain similarities in clinical presentation and confounding factors present in patients who have received kidney allografts, our patient was originally misdiagnosed as having cytomegalovirus retinitis. We emphasise the significance of this diagnostic distinction and the association of this retinal syndrome with renal disease. PMID- 3039416 TI - Evidence that serotonergic projections to the substantia nigra in the rat arise in the dorsal, but not the median, raphe nucleus. AB - Following microinjections of the fluorescent retrograde tracer Fast blue into the substantia nigra, large numbers of retrogradely labeled cells were observed in the dorsal raphe nucleus. Using immunocytochemical techniques it could be demonstrated that the majority of these cells contained serotonin-like immunoreactivity. In contrast, careful examination of the region of the median raphe nucleus revealed no suggestion of a significant projection from the median raphe to the substantia nigra. PMID- 3039417 TI - Postsynaptic inhibition of the frog neuromuscular transmission by prostaglandin E1. AB - Intracellular recordings were made from the frog sartorius muscle end plate. Prostaglandin (PG) E1 (100 nM-10 microM) decreased the amplitude of the end plate potential (EPP). PGE1 decreased the quantal size of EPPs, while it rather increased the quantum content. The frequency of miniature (m) EPPs was not affected by PGE1. PGE1 depressed the acetylcholine (ACh)-induced depolarization, as well as the amplitude of mEPPs. These results suggest that PGE1 decreases the sensitivity of nicotinic ACh receptors at the end plate membrane, resulting in postsynaptic depressions of neuromuscular transmission. PMID- 3039418 TI - Excitatory postsynaptic potentials in neonatal rat sympathetic preganglionic neurons: possible mediation by NMDA receptors. AB - Excitatory postsynaptic potentials (EPSPs) evoked in antidromically identified sympathetic preganglionic neurons (SPNs) and membrane depolarizations induced by N-methyl-D-aspartate (NMDA) applied by pressure ejection were increased by removing Mg ions from the perfusing media and blocked by D-2-amino-5 phosphonovalerate (APV), DL-APV and ketamine. Further, the amplitude of EPSPs and NMDA-induced depolarizations were decreased and increased by membrane hyperpolarization in Krebs solution with and without Mg2+, respectively. These findings indicate that the excitatory amino acid receptor mediating the EPSPs in SPNs may be of the NMDA subtype. PMID- 3039419 TI - No gradient exists between lumbar and ventricular cerebrospinal fluid beta endorphin. AB - In 8 patients, beta-lipotropin (beta-LPH), beta-endorphin (beta-EP), ACTH, protein and chloride concentrations were measured in cerebrospinal fluid (CSF) samples obtained simultaneously from the lumbar space and lateral ventricle. Albumin and IgG levels were significantly higher in lumbar than in ventricular CSF samples while no craniocaudal gradient was observed for neuropeptide concentrations. The importance of molecular weight in determining such a regional distribution is supported also by the similar lumbar and ventricular levels of chlorides. These data would validate the CSF approach through lumbar puncture as a tool for exploring neuropeptides in the central nervous system. PMID- 3039420 TI - Autoradiographic evidence for beta-adrenergic receptors on capsaicin-sensitive primary afferent terminals in rat spinal cord. AB - Specific [3H]dihydroalprenolol binding sites are enriched in the dorsal grey matter of rat spinal cord. Neonatal capsaicin treatment, known to cause a permanent loss of a population of primary afferent neurones, caused a significant loss of specific [3H]dihydroalprenolol binding sites from laminae of the dorsal horn. These results suggest that capsaicin-sensitive primary afferent terminals may bear presynaptic beta-adrenergic receptors. PMID- 3039421 TI - Fast non-cholinergic depolarizing postsynaptic potentials in neurons of rat superior cervical ganglia. AB - After the blockade of cholinergic transmission, stimulation of the preganglionic sympathetic trunk elicited fast depolarizing postsynaptic potentials (PSPs) in rat superior cervical ganglia. At 50 min, their amplitude measured intracellularly was 6.9 +/- 1.7 mV and their duration 25.9 +/- 7.6 ms (mean +/- S.D., n = 9 ganglia). The extracellular electrical activity recorded from the postganglionic internal carotid nerve was monophasic and equal to 4.0 +/- 2.2% of the normal activity (mean +/- S.D., n = 12 ganglia). The effects on these PSPs of some postsynaptic receptor antagonists have been tested. Bicuculline decreased the amplitude of the PSPs as well as that of the monophasic extracellular activity, suggesting that GABA could mediate these non-cholinergic synaptic potentials. PMID- 3039422 TI - Different effects of electrical stimulation of the mesencephalic and pontine reticular formation on the release of dopamine and acetylcholine in the cat caudate nucleus. AB - The effects of unilateral electrical stimulation of the pontine (PRF) and mesencephalic (MRF) reticular formation on the release of acetylcholine (ACh) and of [3H]dopamine continuously synthesised from [3H]tyrosine were examined in both caudate nuclei of halothane-anaesthetised cats implanted with push-pull cannulae. Stimulation of PRF led to a prolonged bilateral increase in the release of [3H]dopamine, whereas a significant reduction in [3H]amine release was observed in the ipsilateral caudate nucleus following stimulation of the MRF. Changes in ACh release were also seen, but they seemed to be independent from those in dopamine release: the release of ACh was enhanced markedly in both caudate nuclei following stimulation of the MRF, whereas a more moderate increase in the release of ACh occurred ipsilaterally following stimulation of the PRF. These data indicate that both the MRF and the PRF are involved in the control of dopaminergic and cholinergic transmission in the basal ganglia. PMID- 3039423 TI - Immunoblockade of response to capsaicin in the rat vas deferens: evidence for the involvement of endogenous calcitonin gene-related peptide. AB - In the rat isolated vas deferens, capsaicin induced a transitory inhibition of the nerve-mediated contractions. This effect was not observed in preparations excised from capsaicin-pretreated rats nor following a first exposure to a high concentration of capsaicin in vitro. Exogenous calcitonin gene-related peptide (CGRP) induced a concentration-related inhibition of the nerve-mediated contractions. A highly avid and specific anti-CGRP serum raised in rabbits against conjugated synthetic rat CGRP inhibited selectively the capsaicin effect. These findings are consistent with the hypothesis that, in this preparation, the specific visceromotor response to capsaicin is brought about by the release of endogenous CGRP from sensory nerves. PMID- 3039424 TI - Androgens stimulate preoptic area Na+,K+-ATPase activity in male rats. AB - This paper describes the effects of castration and testosterone replacement on the Na+,K+-ATPase activity levels of the cerebral cortex (CC), preoptic suprachiasmatic region (POSC) and mediobasal hypothalamus (MBH) in male rats. Na+,K+-ATPase activity was estimated as the ouabain-sensitive fraction of ADP and AMP generation rate, measured by high-pressure liquid chromatography (HPLC) with UV detection, from a standard incubation mixture containing 3 mM ATP. Orchidectomy, performed 4 weeks before sacrifice, decreased ATPase activity of MBH. Testosterone propionate treatment (50 micrograms/day X 2 days) to castrated animals resulted in a 4-fold increase in enzyme activity in the POSC, an effect that might be related to the behavioral effects of androgens. None of the treatments seemed to influence the enzyme activity of the cerebral cortex. PMID- 3039425 TI - Dietary fiber and the risk of cancer. PMID- 3039426 TI - Protocol for hepatitis screening and follow-up of Southeast Asian refugees. AB - Hepatitis B virus (HBV) infection is prevalent among Southeast Asian refugees. HBV infection causes acute and chronic hepatitis and cirrhosis, and is associated with primary hepatocellular carcinoma. Health care providers must be able to make an accurate diagnosis of acute or chronic HBV infection when performing medical evaluations of Southeast Asian refugees. It is essential that appropriate follow up care and teaching regarding infection control be provided. In this article, a protocol is presented that aids interpretation of hepatitis B screening tests. The protocol uses testing for the immunoglobulin M antibody against hepatitis B core antigen (anti-HBc/IgM) to aid differentiation between acute and chronic HBV infection. If testing is interpreted to show the presence of the hepatitis B chronic carrier state, a second protocol is used to guide follow-up testing, treatment and referral of patients. PMID- 3039427 TI - What's new in the diabetic diet. PMID- 3039428 TI - Etiology of infectious mononucleosis. Solving the riddle. PMID- 3039429 TI - A noninvasive technique for monitoring lung vascular permeability in man. AB - Increased microvascular permeability resulting in increased plasma protein extravasation is the hallmark of acute inflammatory oedema and hence radiolabelled proteins can be used to monitor this process. The adult respiratory distress syndrome (ARDS) is characterized by acute inflammatory oedema and thus provides an ideal model for studying this type of oedema in the human lung. A noninvasive technique applicable to the intensive care unit has been developed for monitoring the pulmonary accumulation of the plasma protein transferrin. Transferrin was radiolabelled in vivo with indium-113m and its accumulation was monitored using a portable probe radiation detector. After correcting for changes in intrathoracic blood distribution, by simultaneously monitoring the accumulation of technetium-99m-labelled red blood cells, an index of plasma protein accumulation was calculated. In all patients with established ARDS (n = 10) the index values were greater than 1.0 X 10(-3) min-1 and these were clearly separate from the values of less than 0.5 X 10(-3) min-1 in all healthy volunteers (n = 5; P less than 0.001). The technique can clearly detect raised plasma protein accumulation indices in the lungs of patients with established inflammatory oedema of ARDS and hence may provide a pharmacological tool for the rapid evaluation in these conditions of the effects of drugs (like corticosteroids) which are known to modulate inflammatory oedema. PMID- 3039430 TI - Adrenal scintigraphy in Cushing's syndrome caused by bilateral hyperplasia, adenoma or carcinoma. AB - The results of scintigraphy in 29 patients with Cushing's syndrome were evaluated. It was possible to separate bilateral hyperplasia from unilateral abnormality. In cases of unilateral abnormality correct localization of the tumour was accomplished and it was possible to visualize adrenocortical carcinoma. The many advantages of adrenal scintigraphy are listed and it is concluded that adrenal scintigraphy is the best means of investigation to differentiate between ACTH-dependent and ACTH-independent Cushing's syndrome. PMID- 3039431 TI - Combination chemotherapy of hepatocellular cancer. Comparison of adriamycin + VM 26 + 5-fluorouracil with mAMSA + VM 26 + 5-fluorouracil. AB - Forty-seven patients with hepatocellular cancer were treated in a randomised trial comparing adriamycin + VM 26 + 5-Fluorouracil (5-FU) to mAMSA + VM 26 + 5 FU. Thirteen patients had a partial response to treatment (28%) and another 6 (13%) showed disease stabilisation. There were no significant differences in the response rates between the two treatment regimens. Patients who responded to treatment showed significant prolongation of survival (48 weeks) when compared to non-responders (5 weeks). PMID- 3039432 TI - 201Tl/67Ga uptake ratio in small cell carcinoma of the lung: with special reference to serum neuron-specific enolase and prognosis. AB - 201Tl/67Ga crude uptake ratio (CUR) was measured in 12 patients with small cell carcinoma of the lung (SCCL); it was further compared with serum neuron-specific enolase (NSE) and the actual survival times. A significant inverse correlation was observed between CUR and serum NSE (r = 0.610; p less than 0.05); CUR also correlated with actual survival time (r = 0.605; p less than 0.05). Oat cell-type SCCL having a low CUR and highly raised serum NSE, showed a poorer prognosis than the other subtype which has a high CUR whose serum NSE was not raised as much as in oat cell-type SCCL. This preliminary study suggests that 201Tl/67Ga uptake ratio is a possible prognostic indicator of SCCL. PMID- 3039433 TI - Intradural spinal metastases in patients with posterior fossa brain metastases from various primary cancers. AB - Intradural spinal metastases (ISM) are rare and primarily found associated with certain types of brain tumors like medulloblastoma. Their association with metastases to the brain has been recently described and seems to occur more frequently in posterior fossa lesions. We reviewed patients treated at Massachusetts General Hospital for posterior fossa brain metastases from various primary cancers, and evaluated the risk of concomitant or subsequent ISM. Of 104 patients, 10 developed ISM, with a 1-year actuarial risk of 25%. The risk was not significantly related to age, sex, histology, site of primary tumor or previous therapy. The study suggests that in patients with posterior fossa metastasis, particular attention should be paid to seeding via cerebrospinal fluid and drop metastases to the spinal cord. PMID- 3039434 TI - [Development of the hormonal reaction to stress in the ontogenesis of the baboon Papio hamadryas]. AB - The hormonal reaction of adrenal and gonadal glands of baboons at various ages was studied under 2 hr immobilization stress condition. Concentrations of cortisol, 11-deoxycortisol, 17-hydroxypregnenolone, dehydroepiandrosterone and testosterone were determined by radioimmunoassay in the monkey blood plasma at different times during the stress reaction. A more pronounced reaction of adrenal cortex was shown in 1 year old baboons. The peak of cortisol level in immature monkeys under stress conditions was registered much earlier than in adult monkeys. PMID- 3039435 TI - Effect of prolactin on galactose cataractogenesis. AB - Prolactin has been known to affect the water and electrolyte balance. Because increased lens hydration has been shown to be a common phenomenon in most, if not all types of cataracts, we have been interested in investigating a possible role of prolactin in sugar cataract induction and progression. For this study, we have used morphological and biochemical approaches. The prolactin delivery method involved intraperitoneal implantation of one or more pellets in Sprague-Dawley female rats. Following implantation of the desired number of prolactin or control (nonprolactin) pellets, animals were either fed galactose and lab chow, or lab chow diet. Gross morphological observations of whole lenses, slit-lamp examination of lenses and light microscopic analysis of lens sections showed that in the galactose-fed prolactin group, galactose associated alteration progressed faster and total opacification (mature cataract development) was achieved earlier than in the nonprolactin group. The levels of galactose and dulcitol were higher in the lenses of galactose-fed prolactin treated rats as compared to lenses from nonprolactin (control) rats. No significant difference in lens Na+-K+ ATPase activity between the prolactin and nonprolactin group was observed. Our results indicate that prolactin accelerates galactose-induced cataractogenesis in rats. PMID- 3039436 TI - The use of freeze-dried rib and hydroxylapatite in the treatment of a fracture occurring in a patient with familial facial osteodystrophy. AB - The treatment of repeated pathologic fractures of the mandible has long taxed the ingenuity of surgeons. The complexity of the problem increases with the recognition of a metabolic defect that renders the facial bones hypoplastic. In the case presented, a tentative diagnosis of familial facial osteodystrophy was made at the National Institutes of Health. Since that time, the patient has undergone autogenous onlay grafting, which has completely resorbed. Because of continuous resorption, a number of pathologic fractures have occurred. The most recent fracture was treated by means of a freeze-dried rib, which acted as a floor and as a splint to the fracture site. Hydroxylapatite and a marrow mixture were placed on the floor to gain height, strength, and durability. Subsequently, hydroxylapatite augmentation was performed to gain a more anatomic alveolar ridge. The patient's 12-month follow-up has shown the mandible to be intact with no noticeable change in either vertical or horizontal dimensions. PMID- 3039437 TI - Malignant fibrous histiocytoma of the oral soft tissues. AB - Between 1944 and 1984, six patients with a malignant fibrous histiocytoma of the oral soft tissues were treated at the M.D. Anderson Hospital and Tumor Institute. From the time of the first description of this tumor until 1984, another 10 documented cases of malignant fibrous histiocytoma involving the oral soft tissues were reported in the English-language literature. With respect to these 16 patients (including our 6 cases), the clinicopathologic aspects of malignant fibrous histiocytoma, treatment, and survival figures are discussed. The treatment of choice is early and radical surgery, including neck dissection, since in four of the six patients treated at the M.D. Anderson Hospital and Tumor Institute regional lymph node metastases were found. The response to chemotherapy, except for the response of patients with the inflammatory type of malignant fibrous histiocytoma, and radiotherapy is usually poor. The 2-, 3-, and 5-year survival rates respectively, are, 36%, 30%, and 20%. PMID- 3039438 TI - Gingival granular cell tumor of the newborn: immunoperoxidase investigation with anti-S-100 antiserum. AB - The gingival granular cell tumor (GGCT) of the newborn is a rare, benign tumor, most often observed in the canine area of the maxilla of female infants. The main histologic features are not controversial and involve the occurrence of sheets of granular cells and a prominent vascular component. Conversely, ultrastructural studies agree on limited features and give rise to different hypotheses of mesenchymal origin. S-100 immunoperoxidase investigation of one case shows that, unlike those of granular cell tumor (granular cell myoblastoma) granular cells of GGCT do not react positively for S-100 protein, thus suggesting that the GGCT and the granular cell myoblastoma have a different histogenesis. It confirms also the occurrence of smaller, isolated, spindled, nongranular S-100-positive cells of indeterminate nature, distributed at the periphery of some blood vessels. PMID- 3039439 TI - Surgical approach to tumors arising from the posterior two thirds of the nasal septum. AB - A surgical approach to tumors of the septum is discussed. Two patients who had tumors in the posterior two thirds of the septum are shown in this paper. One had a benign mixed tumor and the other had an extramedullary plasmacytoma in the posterior two thirds of the septum. They underwent resection of the tumors with the septum by lateral rhinotomy. The first case additionally underwent an oval shape resection of the palate. The postoperative profiles of the nose are good. PMID- 3039440 TI - [Islet cell carcinoma]. PMID- 3039441 TI - Carotid artery vasospasm complicating extensive skull base surgery: cause, prevention, and management. AB - Arterial spasm is rarely encountered in the uncomplicated cervical lymphadenectomy. Intense, often dramatic, vasospasm of the internal carotid artery, however, is not infrequently observed in the removal of skull-base lesions. This myogenic reaction is independent of autonomic innervation, occurs more frequently in younger patients, and appears to be due mainly to longitudinal arterial traction and prolonged arterial contact with fresh blood. A case of severe internal carotid artery spasm, which led to a fatal stroke in a young woman who underwent removal of a large glomus jugulare tumor, is presented to emphasize not only the lethal potential of carotid spasm, but intraoperative changes in the character of the artery which suggest the need for immediate spasmolysis. Perioperative guidelines for the prevention and treatment of arterial spasm--including topical and systemic pharmacotherapy and refined surgical techniques--are outlined on the basis of our subsequent experience. PMID- 3039442 TI - [Changes in rickets prevention and conversion of vitamin D administration ("new" Dekristol]. PMID- 3039443 TI - [The mediator of inflammation leukotriene B4]. PMID- 3039444 TI - [Cystic nephroma. Histological and ultrastructural study of a case observed in an adult]. PMID- 3039445 TI - Effect of oxygen on right ventricular performance evaluated by radionuclide angiography in two young patients with chronic lung disease. AB - Pulmonary hypertension is a relatively common complication of chronic lung disease in children that can cause diminished right ventricular performance (RVP) and, eventually, cor pulmonale and heart failure. Since oxygen may decrease pulmonary artery pressure in these patients, we questioned whether RVP would also improve concomitantly. We evaluated the effect of oxygen on RVP in two young hypoxemic patients by radionuclide angiography. A child with bronchopulmonary dysplasia and cor pulmonale who was not clinically in heart failure had acutely better RVP while breathing oxygen and a further improvement after continuous oxygen therapy for 1 year. In a young adult with cystic fibrosis who was suspected of being in heart failure RVP acutely improved when the FIO2 was increased. We conclude that oxygen may improve RVP in hypoxemic patients and speculate that the observation of such improvement may be valuable for the early detection of patients who can benefit from long-term oxygen therapy. PMID- 3039446 TI - Infections in day care. AB - Infections in children in day care are common but can be limited by several measures which include education of providers and staff in standards of hygiene, maintenance of basic techniques of infection control, appropriate use of the physical facilities of the day care facility and maintenance of recommended immunization schedules of children and staff. PMID- 3039447 TI - Magnetic resonance imaging of the brain in childhood herpesvirus infections. AB - We used magnetic resonance imaging (MRI) to evaluate nine children with neurologic disorders caused by infections with members of the herpesvirus family. MRI studies were abnormal in eight children and demonstrated a wide range of central nervous system lesions, including cystic encephalomalacia, ventricular enlargement, cerebral atrophy and focal parenchymal lesions. When compared with conventional computed tomographic scanning, MRI was more sensitive in detecting abnormalities of white matter and in defining the extent of parenchymal lesions. These studies indicate that MRI scans are highly useful in children with herpesvirus infections involving the central nervous system. PMID- 3039448 TI - Effects of potassium bicarbonate on distal nephron Na-K-ATPase in adrenalectomized rabbits. AB - Na-K-ATPase activity in the connecting tubule (CNT) and cortical collecting duct (CCD) has been shown to be influenced by KCl both in the presence and in the absence of aldosterone. To investigate if the aldosterone-independent effect of K+ on Na-K-ATPase can be produced by other K+ salts, we studied the effects of dietary KHCO3 on Na-K-ATPase and ouabain-insensitive Mg-ATPase activities in four nephron segments of adrenalectomized (ADX) rabbits. The segments examined were: the distal convoluted tubule (DCT), CNT, CCD and medullary collecting duct (MCD). All diets were similar in composition except their KHCO3 contents which were 100, 300, 500 and 700 meq/kg in groups 1 to 4 respectively. Increasing KHCO3 in the diet increased K+ excretion (7 X) and urine pH (6.6 to 8.3). Na-K-ATPase activity in the CCD increased greater than 200% as dietary KHCO3 was increased to 700 meq/kg. There was a linear relation between Na-K-ATPase activity in this segment and steady state plasma K+ as well as K+ excretion in the urine. However, Na-K ATPase activity in the CCD was lower in KHCO3-fed ADX rabbits than the KCl-fed animals studied previously under similar conditions. There were no significant differences in Na-K-ATPase activities in DCT, CNT and MCD among the four groups given different KHCO3-diets. It is concluded that dietary intake of KHCO3 can also influence Na-K-ATPase activity in the CCD independent of aldosterone. PMID- 3039449 TI - Evidence for OH-/H+ permeation across the peritubular cell membrane of rat renal proximal tubule in HCO3(-)-free solutions. AB - Membrane potentials and intracellular pH were measured on rat renal proximal tubular cells in vivo to test whether sodium-bicarbonate cotransport across the peritubular cell membrane accepts OH- (or H+ in opposite direction) or whether it requires the CO2, HCO3-, CO3= buffer to operate. It was found that step changes of peritubular pH in nominally HCO3(-)-free and CO2-free solutions produced qualitatively similar initial potential responses and cell pH responses as changes in peritubular HCO3- concentrations. These responses, however, were considerably smaller and they were neither reduced in Na+-free solutions nor inhibited by the stilbene derivative SITS which is known to block Na+ (HCO3-)n cotransport completely. We conclude that the cotransporter requires the CO2, HCO3 , CO3= buffer for its physiological operation but that high rates of OH- or H+ can also be transferred across the peritubular cell membrane in HCO3(-)-free solutions, probably through a separate transport system. PMID- 3039450 TI - Electrophysiological identification of principal and intercalated cells in the rabbit outer medullary collecting duct. AB - Intracellular microelectrode techniques were used together with inhibitors of Na+ transport (amiloride) and H+ transport (acetazolamide and SITS) to identify principal cells and intercalated cells in the outer stripe of the rabbit outer medullary collecting duct. The principal cell (n = 9) had a basolateral membrane voltage (Vbl) of -64.7 +/- 3.2 mV, a fractional resistance of the apical membrane (fRa = Ra/Ra + Rbl) of 0.82 +/- 0.02, and a K+-selective basolateral membrane. Luminal amiloride hyperpolarized Vbl by 10.3 +/- 2.1 mV and increased fRa to near unity (n = 7). Bath acetazolamide and SITS were without effect on these parameters. The intercalated cell (n = 5) had a Vbl of -25.0 +/- 3.2 mV, a fRa of 0.99 +/- 0.01, and a Cl(-)-selective basolateral membrane. Bath acetazolamide or SITS hyperpolarized Vbl by 26.4 +/- 8.2 mV. Luminal amiloride did not alter Vbl of this cell. The differential effects of the inhibitors also indicate that the principal and intercalated cells are probably not directly coupled electrically. PMID- 3039451 TI - Forskolin inhibition of hexose transport in cardiomyocytes. AB - The effects of insulin, forskolin, isoproterenol, and epinephrine on 3-O methylglucose (hexose) transport and cell cyclic AMP levels were determined in adult rat cardiomyocytes. Insulin stimulated hexose transport in these cells an average of 2.5-fold. Initial hexose transport rates at 1 mM hexose were 3.75 X 10(-2) nmol/mg cell protein/second in the absence of insulin, and 8.25 X 10(-2) nmol/mg cell protein/second in the presence of 12.3 microM insulin. Forskolin at 5 microM nearly abolished hexose transport within 3 s of exposure, but did not increase cell cyclic AMP concentrations within 9 s. The apparent Ki for hexose transport inhibition was about 0.3 microM forskolin. Epinephrine and isoproterenol at 50 microM increased cell cyclic AMP 4-fold during 9 s exposure, but did not affect hexose transport. Treatment of cells with these catecholamines of forskolin for up to 99 s increased cell cyclic AMP, but only forskolin inhibited hexose transport. We conclude from these results that forskolin acts on hexose transport independent of its action on adenyl cyclase, and that cyclic AMP does not inhibit or stimulate hexose transport. PMID- 3039452 TI - Effects of prolactin on Na-K-ATPase activity along the rat nephron. AB - To test prolactin (PRL) action on osmoregulation in mammals, we evaluated in the rat the effect of this hormone on a major enzyme in renal regulation of water and electrolyte: renal Na-K-ATPase. Enzyme activity was determined by cytochemistry in medullary ascending limb (MAL) and distal convoluted tubule (DCT) from rats treated either by bromocriptine, or by PRL. Three hours after a bromocriptine injection (0.1 mg/100 g IP) a significant decrease of Na-K-ATPase activity is observed in both MAL (80% of control values, p less than 0.001) and DCT (78% p less than 0.01). Reciprocally, a significant (p less than 0.001) increase in enzyme activity is induced 3 h after a single PRL injection (140 micrograms/100 g IM), in both segments (MAL: 165%, DCT: 172% of control activities) and persists 6 h after the injection (MAL: 130%, DCT: 118%). Na-K-ATPase activity was correlated to plasma PRL levels (r = 0.78 in DCT, r = 0.89 in MAL). A direct effect of PRL on the tubule is suggested by results from experiments in which PRL, at various concentrations, is added in vitro on renal slices before Na-K-ATPase activity measurements. The increase in Na-K-ATPase activity exhibits a log-dose dependency with PRL concentration (p less than 0.01) and is still observed when AVP antagonist is added before PRL incubation, ruling out the possible role of AVP contamination of PRL. These results suggest a direct effect of PRL on renal Na-K ATPase in MAL and DCT. PMID- 3039453 TI - Structural and functional response of the isolated toad skin to mucosal lithium. AB - Structural and functional changes induced by long-term Li-exposure of the outer surface of toad skin was studied. Electron microscopy revealed that total Na by Li replacement in the outer compartment of short-circuited toad skin promotes a conspicuous cellular damage expressed as focal swollen cells with altered intercellular spaces and nuclear morphology. Short-circuit current (SCC) decreases by about 70% over the first 60 min after 115 mmol/l Li-exposure. An amiloride sensitive transepithelial Li transport remains intact over a further 150 min despite the epithelial damage, indicating that the pathways across the apical barrier are functioning. Increase of the paracellular permeability is detected by elevation of Na-efflux. Partial 50% or 10% Na by Li replacements induce minor structural alterations and are not sufficient to trigger appreciable Na-efflux and SCC alterations. Therefore, low Li concentrations are less effective than 115 mmol/l in promoting morphofunctional responses. Although ouabain and Li reduce the SCC, ouabain does not promote structural lesions, showing that Li-inhibition of Na transport by itself is not responsible for the observed morphological alterations. In the light of this study, Li utilization as a tool to investigate transepithelial Na transport requires careful judgement. PMID- 3039454 TI - Evidence for a Na+/H+ exchanger at the basolateral membranes of the isolated frog skin epithelium: effect of amiloride analogues. AB - We have investigated the possible existence of a Na+/H+ ion exchanger in the frog skin epithelium by using isotopic methods and two amiloride analogues:5-(N-ethyl N-isopropyl)-amiloride (EIPA) and phenamil. We found phenamil to be a specific blocker of sodium entry to its cellular transport compartment since it inhibited both the transepithelial Na+ influxes (J13) with a K1 of 4 X 10(-7) mol/l and the Na+ pool (control: 77 +/- 4 neq X h-1 X cm-2; phenamil: 21 +/- 1 neq X h-1 X cm 2). On the contrary EIPA (10(-5) mol/l) had no effect on J13 nor on the apical Na+ conductance. Acidification of the epithelium by passing from a normal Ringer (25 mmol/l HCO3-, 5% CO2, pH 7.34) to a HCO3(-)-free Ringer (5% CO2, pH 6.20) while blocking the Na+ conductance with phenamil, produced a large stimulation of Na+ influxes exclusively across the basolateral membranes (J32), after return to a normal Ringer (J32 = 706 +/- 76 and 1635 +/- 199 neq X h-1 X cm-2 in control and acid-loaded epithelia respectively). The stimulation of J32 was initiated when the epithelia were acid-loaded with Ringer of pH lower than 6.90 and was blocked by amiloride (KI = 7 X 10(-6) mol/l) and EIPA (KI = 5 X 10(-7) mol/l) whereas phenamil had no effect. In Na+-loaded epithelia (ouabain treated) the Na+ efflux across the basolateral membranes was stimulated by an inwardly directed proton gradient and was blocked by EIPA (10(-5) mol/l) or amiloride (10(-4) mol/l), a result suggesting reversibility of the mechanism. We conclude that a Na+ permeability mediated by a Na+/H+ ion exchanger exists in the basolateral membranes, which is stimulated by intracellular acidification and is sensitive to amiloride or EIPA. This exchanger is proposed to be involved in intracellular pH regulation. PMID- 3039455 TI - Dibutyryl cyclic AMP inhibits transport dependent QO2 in cells isolated from the rabbit medullary ascending limb. AB - Because the medullary thick ascending limb of the loop of Henle is the target of several polypeptide hormones that stimulate adenyl cyclase in this nephron segment, we examined the effects of cyclic AMP on thick ascending limb of the loop of Henle cells isolated by enzymatic digestion and density gradient centrifugation from the outer medulla of the rabbit kidney. The functional parameter that was measured was transport dependent oxygen consumption. Oxygen consumption was measured using a polarographic oxygen electrode in a constant temperature chamber. We found that dibutyryl cyclic AMP inhibited oxygen consumption in a dose dependent way. Maximal inhibition was observed at a concentration of 10(-5) M. The effect of dibutyryl cyclic AMP was not present in the absence of either sodium, chloride or both implying that its effect is restricted to the sodium and chloride dependent oxygen consumption. The effect of dibutyryl cyclic AMP was additive to that of furosemide 10(-4) M while that of furosemide was not additive to that of cyclic AMP suggesting that the site of action of cyclic AMP is distal to that of furosemide. The effect of dibutyryl cyclic AMP was not additive to that of ouabain and was absent in cells where oxygen consumption was stimulated with amphotericin B in the absence of chloride indicating that it has no effect on Na-K-ATPase.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3039456 TI - [Responses to radiation of Ehrlich ascites tumor cells in the presence of YM 08310 (=WR-2721) and/or Ro-07-0582 (in vivo, in vitro assay)]. PMID- 3039457 TI - [The role of radiation therapy in the treatment of small cell carcinoma of the lung--with a special reference to the optimal combination timing within chemotherapy and total radiation doses]. PMID- 3039458 TI - [Dose estimation by electron spin resonance (ESR) of the tooth enamel of radiologic technologists]. PMID- 3039459 TI - The use of permanganate as a sequencing reagent for identification of 5 methylcytosine residues in DNA. AB - The use of permanganate as a reagent for DNA sequencing by chemical degradation has been studied with respect to its specificity for 5-methylcytosine residues. At weakly acidic pH and room temperature, 0.2 mM potassium permanganate reacts preferentially with thymine, 5-methylcytosine, and to a lesser extent with purine residues, while cytosine remains essentially intact. Permanganate oxidation is, therefore, a suitable DNA sequencing reaction for positive discrimination between 5-methylcytosine and unmethylated cytosine. PMID- 3039460 TI - Site-specific DNA binding of nuclear factor I: effect of the spacer region. AB - Nuclear factor I (NFI) is a site-specific DNA binding protein required for the replication of adenovirus type 2 DNA in vitro and in vivo. To study sequence requirements for the interaction of NFI with DNA, we have measured the binding of the protein to a variety of synthetic sites. Binding sites for NFI (FIB sites) were previously shown to contain a consensus sequence composed of 2 motifs, TGG (Motif 1), and GCCAA (Motif 2), separated by a 6 or 7bp spacer region. To assess conserved sequences in the spacer region and flanking sequences which affect NFI binding, we have isolated clones from oligonucleotide libraries that contain the two motifs flanked by 3 degenerate nucleotides and separated by degenerate spacer regions of 6 or 7 nucleotides. With a 6bp spacer region, a strong bias exists for a C or A residue in the first position of the spacer. Sites with a 7bp spacer region contain a G and C or A residue at the first and second positions, respectively, of the spacer, but also possess conserved residues at other positions of the site. PMID- 3039461 TI - Initiation signals for the conversion of single stranded to double stranded DNA forms in the streptococcal plasmid pLS1. AB - We have characterized a region in the streptococcal plasmid pLS1 located between nucleotides 4103 and 4218 which is a signal involved in the conversion of single stranded intermediates of replication to double stranded plasmid forms. This region has a large axis of dyad symmetry resulting in the formation of a secondary structure as revealed by the location of endonuclease S1-cleavage sites in supercoiled covalently closed circular pLS1 DNA. Deletions affecting this region caused a fivefold reduction in plasmid copy number, plasmid instability and the accumulation of single-stranded DNA intermediates. The conversion signal of pLS1 has homologues in other staphylococcal plasmids, sharing a consensus sequence located in the loop of the signal. Computer assisted analysis showed that the signal detected in pLS1 has a high degree of homology with the complementary strand origin of the Escherichia coli single stranded bacteriophages phi X174 and M13. PMID- 3039462 TI - Putative promoter elements for the ribosomal RNA genes of the thermoacidophilic archaebacterium Sulfolobus sp. strain B12. AB - In Sulfolobus sp. strain B12, single-copy genes encode the three ribosomal RNAs. The genes for the 16S rRNA and for the 23S rRNA are closely linked but separated from the 5S rRNA gene. Transcription of the 16S/23S rRNA gene cluster starts 139 nucleotides upstream of the 5'-end of mature 16S rRNA. For the 5S rRNA gene the point of transcription initiation coincides with the 5'-end of mature 5S rRNA. The comparison of the upstream regions for these transcriptional start sites shows the presence of a completely conserved trinucleotide sequence around the point of transcription initiation and a completely conserved octanucleotide sequence about 22 nucleotides upstream of it. These sequences are only moderately homologous to putative promoter elements for stable RNA genes in the closely related archaebacterium Thermoproteus tenax (1), but they are very similar to corresponding sequences in the distantly related archaebacterium Methanococcus vannielii (2). The consensus sequence found for Sulfolobus and Methanococcus could therefore constitute the archetype of an archaebacterial promoter for stable RNA genes. PMID- 3039463 TI - Characterization of two types of histone H2B genes from macronuclei of Tetrahymena thermophila. AB - Two histone H2B gene clones were isolated from macronuclei of Tetrahymena thermophila. Nucleotide sequences of the two clones were highly homologous within the coding region but not in the noncoding region. Comparison of the deduced amino acid sequences between the two clones showed three differences in a total of 121 amino acids. Each of the two clones contained a TAA triplet within the coding region, which appeared to code for a glutamine residue. To demonstrate the existence of histone mRNA containing UAA triplet, nuclease P1 protection mapping using total cellular RNA and nucleotide sequencing of primer extension products were carried out. The results clearly indicated that two cloned histone H2B genes were transcribed, giving rise to the major histone H2B mRNAs with a UAA triplet sequence in frame. The tentative 5'- and 3'-ends of histone H2B mRNAs were determined. PMID- 3039465 TI - The anomalous gel migration of a stable cruciform: temperature and salt dependence, and some comparisons with curved DNA. AB - We have made an analysis of the gel electrophoretic properties of a pseudo cruciform fragment, a linear DNA molecule containing a stable cruciform. The migration of this construct was analysed in polyacrylamide gels at a various temperatures in the range 5 degrees to 55 degrees C, and in the presence of NaCl, MgCl2 or ethidium bromide. The magnitude of the anomalous migration (retardation) was almost temperature independent up to 40 degrees C, but decreased strongly beyond this point, extrapolating to normal migration at 70 degrees C. Addition of salts reduced the anomaly. This took the form of a continuous reduction in anomalous migration with the addition of NaCl up to 60 mM, while with MgCl2 there was a sharp reduction in the anomaly to a constant value which is reached by 10 mM. Under these conditions, moreover, the migration of the fragment became almost temperature-independent over the entire range. These results have been interpreted to reflect the influence of ion binding at the four-way junction on the relative disposition of the cruciform arms. The detailed electrophoretic properties of the pseudo-cruciform are in marked contrast to those of sequence directed curved DNA fragments. In particular, the response to the addition of 1 microgram/ml ethidium bromide offers a convenient method for distinguishing between anomalous retardation arising from curvature (greatly reduced anomaly) or a cruciform junction (enhanced anomaly). PMID- 3039464 TI - The human beta-globin gene contains a downstream developmental specific enhancer. AB - The human beta-globin gene is part of a multigene family and is expressed specifically in adult human erythroid tissue (for review, 1). When the human beta globin is introduced into fertilized mouse eggs, it is first activated in foetal liver and remains expressed in adult erythroid tissues. It therefore mimicks the pattern of expression of its murine counterpart. It has previously been shown in tissue culture and transgenic mice that sequences downstream from the beta-globin promoter are involved in this regulation. We now show that at least part of these sequences are located 0.5-1.2kb downstream from the poly A addition site and constitute a transcriptional enhancer element that is erythroid and developmental specific. PMID- 3039466 TI - Two-dimensional NMR investigation of a bent DNA fragment: assignment of the proton resonances and preliminary structure analysis. AB - The resonances of all the non-exchangeable protons (except 5'H and 5"H) of d(CGAAAAATCGG) + d(CCGATTTTTCG), a putatively bent DNA duplex, have been assigned using 1H two-dimensional nuclear magnetic resonance methods. The nuclear Overhauser effect data indicate an overall B-form structure for this double helical DNA undecamer. However, several features of the NMR data such as some unusually weak C8/C6 proton to C1' proton NOE cross-peaks, the presence of relatively intense C2H to C1'H NOE cross-peaks, and unusual chemical shifts of some 2", 2', and 1' protons suggest a substantial perturbation of the helix structure at the junctions and along the length of the tract of A residues. These structural deviations are considered in terms of models of DNA bending. PMID- 3039467 TI - Clustered alternative splice sites in Epstein-Barr virus RNAs. PMID- 3039468 TI - Nucleotide sequence analysis of long terminal repeats of leukemogenic and non leukemogenic MCF MuLVs. PMID- 3039469 TI - Colonic carcinogenesis: the microbial feast or famine mechanism. AB - A mechanism is presented which suggests that high-fat, high-protein, low-fiber diets can cause an unfavorable microbial environment in the human colon which predisposes some individuals towards large bowel diseases. The digesta leaving the ileum on high-fat, high-protein, low-fiber diets has a high proportion of mucins, malabsorbed carbohydrates and proteins, bile acids, and sloughed epithelial cells. The irregular (pulsatile) emptying of rapidly fermentable ileal digesta into the colon causes a massive surge in microbial activity. The sudden availability of rapidly fermentable substrate generates a large microbial population in the exponential growth phase that soon depletes its substrate. For microorganisms to perpetuate until the next high-fat, high-protein, low-fiber meal propels ileal digesta into the colon, they must induce enzymes to ferment dying or dead microbes (cannibalism) in addition to colonic epithelial mucosa and mucins. As the carbohydrate-to-nitrogen ratio of the colonic contents decreases, the fermentation becomes more proteolytic and subacute levels of fermentation products such as ammonia may be generated. Carcinogens are concentrated within a small colonic mass and the probability of precancerous lesions and polyps developing in the colonic mucosa is directly related to the severity, frequency, and duration of these microbial "feast or famine" situations in the colon. PMID- 3039470 TI - Intestinal carcinogenesis and dietary fibers: the influence of cellulose or Fybogel chronically given after exposure to DMH. AB - This study was initiated to analyze the effect of a) two characterized vegetal fibers [i.e., a polysaccharide (cellulose) and a mucilaginous substance (Fybogel)], which were b) added at two concentrations (5% and 15% wt/wt), c) as constituents of low (5% wt/wt) and high (20% wt/wt) fat isocaloric diets d) given chronically to rats one week after the administration of 1,2-dimethylhydrazine (DMH); DMH had previously been injected once a week for 15 weeks to induce intestinal carcinogenesis. The dietary consumption, the body weight, and the fecal outflow showed a similar and regular evolution for the rats of all experimental groups; the exception was those receiving the 20% lipids-15% Fybogel diet. That specific diet caused a decrease in body weight concomitant with an increase in dietary consumption and in fecal outflow. The variation in fecal outflow depended on fat and fiber concentrations. The mucilage was more degraded, in absolute and relative amount, than was cellulose when this polysaccharide was included at a 20% lipid diet. Concerning the effect of these two fibers on intestinal carcinogenesis, Fybogel showed an anticarcinogenic property, whereas cellulose did not. The inhibitory activity of Fybogel was on the incidence of intestinal and colonic tumors as well as on the colonic tumor yield. Moreover, it slowed down the rate of colonic formation. PMID- 3039471 TI - [Analysis of the treatment of familial polyposis of the large intestine (1962 1985)]. PMID- 3039472 TI - [A typical clinical forms of familial polyposis of the large intestine]. PMID- 3039473 TI - [Role of membrane receptors in the pathogenesis of epilepsy and in the mechanism of action of anti-epileptic drugs]. PMID- 3039474 TI - [Cyclic nucleotides and prostaglandins E in the supernatant of cell cultures of myeloblastic, myelocytic and lymphocytic leukemias]. PMID- 3039475 TI - Positron emission tomographic evaluations in the diagnosis and therapy of multifocal glioblastoma. Report of a pediatric case. AB - A pediatric case of multifocal glioblastoma is reported with special reference to positron emission tomography (PET) evaluations in the diagnosis and therapy. A PET scan employing 15O and 18F positron tracers disclosed a right thalamic lesion high in regional cerebral blood flow, cerebral blood volume and metabolic glucose rate, suggesting a malignant tumor. In contrast, a left frontal lesion with low regional cerebral blood flow, cerebral blood volume and metabolic glucose rate was presumed to be rather hypodynamic and hypometabolic. Histological examination of these isolated masses showed both to be glioblastomas, and, as predicted by the PET findings, there were minor histological distinctions. In repeated PET scans following postoperative combined radiochemotherapy, metabolic glucose rate values at the thalamic tumor decreased to about 50% the levels before therapy, although there was no distinguishable change in tumor size on the concurrent computerized tomography scans. This therapeutic efficacy on tumor metabolism was highly consistent with the period of clinical relief. PMID- 3039476 TI - Pediatric neurosurgical implications of the amniotic band disruption complex. Case reports and review of the literature. AB - Defects caused by the amniotic band disruption complex (ABDC) may vary from simple malformations caused by digital constriction to major scalp, craniofacial, and visceral malformations. ABDC may cause 7-14% of stillbirths. The etiology is unclear, but the most commonly accepted mechanism involves rupture of the amnion followed by fetal malformation, deformation, and compression. This mechanism does not adequately explain all anomalies such as hydrocephalus and holoprosencephaly that are seen in the ABDC. The use of prenatal ultrasound has allowed the diagnosis of the ABDC in utero. Since 1980, four children with the ABDC who required neurosurgical intervention were seen at the University of California, San Francisco; the presentation and subsequent surgical treatment of 2 of these children are discussed. A combined craniofacial team approach to the management of these children can maximize reconstructive and neurologic outcome. PMID- 3039477 TI - Management of children with primitive neuroectodermal tumors of the posterior fossa/medulloblastoma. AB - Primitive neuroectodermal tumors of the posterior fossa/medulloblastoma (PNET-MB) are the most common malignant primary central nervous system tumors of childhood. Current approaches to therapy for children with PNET-MB are illustrative of both the advances which have been made in management of childhood brain tumors and the areas where increased understanding is needed. With the optimal use of surgery and radiotherapy, the majority of children with PNET-MB can be expected to be alive and free of disease 5 years after diagnosis. Based on various factors, including the age of patient, the extent of the disease at the time of diagnosis and histological parameters, it is possible to stratify patients with PNET-MB into 'risk' groups. It is unclear whether preoperative or postoperative factors are most predictive of outcome. Patients with factors predictive of a higher likelihood of disease relapse seem to benefit from treatment with chemotherapy. The amount of radiotherapy and the type and amount of chemotherapy which is most efficacious in controlling the disease has yet to be determined. Children surviving PNET-MB are at high risk for endocrinologic and neuropsychologic sequelae. Multiple prospective treatment protocols are now underway, attempting to determine which therapy is best at controlling disease without causing severe long-term damage. A multidisciplinary approach is needed for the treatment of children with PNET-MB. PMID- 3039478 TI - Effect of dietary acidification on kidney damage induced in immature chickens by excess calcium and infectious bronchitis virus. AB - Experiments were designed to evaluate the effect of dietary acidification on the development of kidney lesions induced by excess dietary calcium (Ca) and Gray strain infectious bronchitis virus (IBV). Specific pathogen-free (SPF) chicks and SPF chicks inoculated with Gray strain IBV were fed one of three diets: a commercial pullet grower ration (1% Ca); a commercial layer ration (3.25% Ca); or layer ration plus .5% ammonium chloride (acidified layer ration). Gray strain IBV significantly reduced total kidney weights in males, reduced total kidney weight as a percentage of body weight in males, increased the number of gross kidney lesions, and decreased the number of filtering nephrons when compared with uninoculated birds when both groups were fed the grower ration. The layer ration induced a 60% incidence of kidney lesions, caused a significant increase in kidney weight asymmetry ratios, and caused a 25% reduction in the number of filtering nephrons. Acidifying the layer ration significantly reduced the incidence of gross kidney lesions and reduced kidney weight asymmetry ratios, but did not prevent Ca-induced reductions in filtering nephrons. PMID- 3039479 TI - Eggshell quality as influenced by sodium bicarbonate, calcium source, and photoperiod. AB - The effects of Ca source (limestone or a mixture of one-third limestone and two thirds oyster shell), NaHCO3 (0 or .5%) and feeding photoperiod (16 or 24 hr) were studied in a 16-week experiment involving 240 Hy-Line W-36 Leghorn hens, 25 weeks of age. Neither egg production (percent hen-day and egg mass, kg egg/bird per 16 weeks) nor feed conversion ratio (kg feed per kg egg) were significantly (P greater than .05) affected by dietary treatment or photoperiod. However, extending the photoperiod to 24 hr significantly (P less than .05) increased feed consumption with a consequent increase in egg weight (P less than .01). Feeding oyster shell in combination with limestone significantly (P less than .05) improved specific gravity of eggs, whereas dietary NaHCO3 significantly (P less than .01) improved elasticity of the egg shell as measured by deformation. Eggshell quality was improved by increasing the photoperiod to 24 hr and was most pronounced when hens were fed diets supplemented with .5% NaHCO3 and limestone as the only source of Ca. PMID- 3039480 TI - Antibodies to avian infectious bronchitis virus in Pakistani chickens. AB - Using the hemagglutination inhibition test, sera from 20 diseased and 3 apparently healthy chicken flocks in Pakistan were examined for antibodies to four types of avian infectious bronchitis virus (AIBV). These flocks were not vaccinated against AIBV. Of the 900 serum samples from diseased flocks, 78 (8.7%) had antibodies to the Massachusetts (M41) type, 23 (2.6%) to the Arkansas type and 20 (2.2%) to the Connecticut type of AIBV. None had antibodies to the JMK type. None of the sera (n = 100) from apparently healthy layers was positive to any of the four AIBV types tested. Some of the birds with antibody titers exhibited no signs of illness except that they laid pale colored eggs or had a history of poor hatchability. These results indicate that at least three AIBV types are circulating in chicken flocks in Pakistan. PMID- 3039481 TI - Effects of some antihypertensive drugs on cutaneous blood flow and inflammatory skin responses following allergen challenge in guinea pigs. AB - The effects of the antihypertensive drugs clonidine, prazosin, and MK 422 (the active parent diacid of enalapril) were studied on the cutaneous blood flow and allergen evoked inflammatory skin reactions in ovalbumin sensitized guinea pigs. The hypotensive effect of the drugs did not significantly change the basal cutaneous blood flow at the time of allergen challenge. MK 422 (0.02 mg/kg) markedly enhanced the wheal and flare reaction following allergen challenge, whereas clonidine (0.005 and 0.05 mg/kg) inhibited the inflammatory response. Prazosin (0.03 mg/kg) did not significantly influence the wheal and flare reaction. Our results indicate that some antihypertensive drugs (clonidine) could be beneficial to antihypertensive patients with inflammatory diseases, while others (ACE-inhibitors) may enhance their inflammatory disorders. PMID- 3039482 TI - Antagonist effects of the enantiomers of 3-PPP towards alpha 1-adrenoceptors coupled to inositol phospholipid breakdown in the rat cerebral cortex. AB - The effects of the two enantiomers of 3-PPP upon alpha 1-adrenergic and muscarinic receptors coupled to the inositol phospholipid (PI) breakdown response have been investigated. 3-PPP(-) and 3-PPP(+) were found to antagonize the noradrenaline (10 microM)-stimulated PI breakdown in rat cerebral cortical miniprisms with IC50 values of 18 and 61 microM, respectively. The dopamine receptor antagonists haloperidol and raclopride were also antagonists, with IC50 values of 0.4 and 25 microM, respectively. 3-PPP(-) and raclopride were found further to act as competitive antagonists, with pA2 values of 6.03 and 5.44, respectively. 3-PPP(-), 3-PPP(+) and haloperidol also antagonized the muscarinic receptor-mediated carbachol (50 microM)-stimulated PI breakdown in cortical miniprisms, albeit at high concentrations (IC50 values of 91, 170 and 28 microM, respectively) whereas raclopride produced only 24% inhibition at the highest concentration tested (100 microM). PMID- 3039483 TI - Endothelial-smooth muscle cell interactions during atherosclerosis. PMID- 3039484 TI - [Danger of false cytologic interpretation in cytostatic pneumopathy]. PMID- 3039485 TI - HLA expression by trophoblast of invasive moles. AB - HLA expression by the trophoblast in invasive hydatidiform mole was analysed by immunoperoxidase staining. In the invading villi of an invasive mole, villous trophoblast, both syncytiotrophoblast and cytotrophoblast, failed to show a positive reaction for HLA-A, -B and -C and HLA-DR. By contrast, extravillous trophoblast showed an intense reaction for HLA-A, -B and -C. The distribution of HLA antigens in the invading villi was the same as in the non-invading villi, and the antigens were also indistinguishable from those noted in non-invasive hydatidiform moles. The histopathology of invasive mole may suggest that it is a malignant neoplasm. This immunohistochemical study, however, lends support to the current view that invasive mole is a variant of a benign hydatidiform mole rather than a form of malignant trophoblastic disease. PMID- 3039486 TI - [Sweet's syndrome. 2 cases of probable infectious origin]. PMID- 3039487 TI - [The use of immobilized enzymes for the synthesis of nucleosides and nucleotides including the ones labeled with radioactive isotopes]. AB - Nucleoside phosphorylases from E. coli immobilized in polyacrylamide gel were used for production of nucleosides from nitrous bases during the transglucosylase reaction. Phosphorylation of nucleosides resulted in the formation of 5' nucleotides was performed in the presence of carrot nucleoside phosphotransferase immobilized in polyacrylamide gel. Using 14C-labelled nitrous bases as starting substrates, labelled nucleosides and nucleotides can be obtained with the 75-80% yield that have radioactive purity of 95-99%. The stability of the immobilized enzymes was being studied under the conditions of the reactions using them in the batch and plug flow reactors. PMID- 3039488 TI - [Neuroendocrine regulation of immunogenesis in patients with pulmonary tuberculosis and alcoholism]. PMID- 3039489 TI - Human leukocyte collagenase: recent biochemical findings. PMID- 3039490 TI - Collagenolytic enzymes in periodontal diseases. PMID- 3039491 TI - Polymorphonuclear leukocytes and their functions. PMID- 3039492 TI - Characterization of a mammalian mutant with a camptothecin-resistant DNA topoisomerase I. AB - DNA topoisomerase I was purified to near homogeneity from a clonal line of human lymphoblastic leukemia cells, RPMI 8402, that is resistant to camptothecin, a cytotoxic alkaloid from Camptotheca acuminata, and compared with that of the parent wild-type cells. As assayed by relaxation of the supercoiled plasmid DNA and by formation of enzyme-linked DNA breaks, the purified enzyme from the resistant cells was shown to be greater than 125-fold as resistant to camptothecin as the wild-type enzyme, comparable to a cellular resistance index of about 300. Therefore, the cellular resistance appears to be due to the resistance of the enzyme. The amount of the immunoreactive enzyme protein in whole extract appeared to be reduced to less than half that of the wild-type enzyme. These results establish that DNA topoisomerase I is the cellular target of camptothecin and that DNA topoisomerase I is essential for the survival of mammalian cells. PMID- 3039493 TI - Multiple repressor binding sites in the genome of bacteriophage P1. AB - After digestion of bacteriophage P1 DNA with EcoRI in the presence of P1 repressor, 6 repressor binding sites were identified in 5 of 26 EcoRI fragments. Binding sites were localized by the decreased mobility of DNA fragment-repressor complexes during electrophoresis and by DNase protection ("footprinting") analysis. The repressor binding sites, or operators, comprise a 17-base-pair-long consensus sequence lacking symmetrical elements. Three operators can be related to known genes, whereas the function of the others is still unknown. The mutant P1 bac, rendering ban expression constitutive, is identified as an operator constitutive mutation of the ban operon. PMID- 3039494 TI - Identification of a cell-specific transcriptional enhancer in the first intron of the mouse alpha 2 (type I) collagen gene. AB - A transcriptional enhancer has been identified in the first intron of the mouse alpha 2 (type I) collagen gene in a region between +418 and +1524 base pairs from the transcriptional start site. The enhancer functions both when it is located 5' and 3' to the promoter that it activates and is independent of the orientation of the element. The enhancer stimulates both the homologous alpha 2 type I [alpha 2(I)] collagen promoter and the heterologous early simian virus 40 promoter. In transient expression experiments, enhancer-dependent transcription from the alpha 2(I) collagen promoter utilizes the same transcriptional start site as the one used in the endogenous alpha 2(I) collagen gene. The enhancer activates transcription at a distance of at least 3 kilobase pairs from the transcriptional start site. The alpha 2(I) collagen enhancer displays cell specificity, since it is functional in NIH 3T3 fibroblasts but completely inactive in a lymphoid cell line, in contrast to two immunoglobulin gene enhancers that show the opposite behavior. We find several areas of sequence homology with viral enhancers, particularly the enhancer of simian virus 40. PMID- 3039495 TI - Bacteriorhodopsin mutants containing single tyrosine to phenylalanine substitutions are all active in proton translocation. AB - To study the possible role of the tyrosine residues in proton translocation by bacteriorhodopsin, we have replaced these residues individually by phenylalanine. The required codon changes were introduced in the bacterioopsin gene by replacement of appropriate restriction fragments by synthetic counterparts containing the desired nucleotide changes. The denatured opsin polypeptides obtained by expression of the mutant genes in Escherichia coli were purified and treated with a mixture of detergents, phospholipids, and retinal in a previously established renaturation procedure. All of the mutant proteins folded to regenerate bacteriorhodopsin-like chromophores. Three mutants with tyrosine to phenylalanine substitutions at positions 57, 83, and 185 regenerated the chromophore more slowly than the wild-type protein, and two of these mutants, Phe 57 and -83, showed slightly blue-shifted chromophores. When reconstituted into liposomes all of the mutant proteins with single Tyr----Phe substitutions pumped protons at rates and levels comparable to those of the wild-type bacteriorhodopsin. We conclude that single substitutions of tyrosine by phenylalanine do not affect folding, retinal binding, or light-dependent proton pumping in bacteriorhodopsin. PMID- 3039496 TI - Molecular cloning of rat renin cDNA and its gene. AB - Renin (EC 3.4.23.15) is an aspartyl protease that cleaves its only known substrate, angiotensinogen, to release the vasopressor hormone angiotensin. We have isolated full-length cDNAs for renin from a rat kidney cDNA library. The cDNAs are complementary to a 1434-nucleotide rat kidney mRNA that encodes preprorenin, the 402-amino acid precursor of renin. This rat cDNA was used to isolate the complete copy of a renin gene from a rat genomic library, and a comparison of this genomic clone with rat genomic DNA showed that renin is a single-copy gene in the rat. Rat renin is 85% identical to one mouse renin isozyme (renin-1) and 81% identical to the second mouse renin isozyme (renin-2), suggesting that the duplication of the mouse renin genes is a more recent event than the speciation of rats and mice. Analyses of rat, human, and mouse renin sequences revealed that the potential to form two-chain renin is apparently peculiar to mouse renin, and the expression of a tenth exon (which results in a three-amino acid insertion) is observed only in the human renin gene. PMID- 3039497 TI - Agents that elevate the concentration of cAMP in platelets inhibit the formation of a NaDodSO4-resistant complex between thrombin and a 40-kDa protein. AB - We studied the influence of prostaglandin E1 and theophylline on the ability of rabbit or human platelets to form NaDodSO4-resistant complexes between 125I labeled thrombin and a platelet protein of approximately equal to 40 kDa. Control platelets formed two types of these complexes, one that sedimented with the platelets and one that was found in the suspension medium. There were 30-40 sedimentable complexes per platelet and about three times this number of soluble complexes. Pretreatment of rabbit or human platelets with prostaglandin E1 and theophylline decreased the formation of both types of complex by as much as 60 80%. The pretreatment was particularly effective when low doses of thrombin were used. When added to control platelets, such doses of thrombin caused the formation of sedimentable complexes and the concomitant formation of inositol trisphosphate. However, when added to platelets that had been pretreated with prostaglandin E1 and theophylline, low doses of thrombin had little or no effect on either reaction. PMID- 3039498 TI - Adenosine triphosphate stimulates inositol phospholipid metabolism and prostacyclin formation in adrenal medullary endothelial cells by means of P2 purinergic receptors. AB - In the adrenal medulla, chromaffin cells secrete high concentrations of catecholamines, ATP, peptides and other factors that must pass through an endothelial cell barrier to enter the bloodstream. We have measured the effect of several of these chromaffin cell secretory products on cultured bovine adrenal medullary endothelial cells and have found that only ATP stimulates prostacyclin formation. The stimulation of prostacyclin formation by ATP coincides with the metabolism of inositol phospholipids and the accumulation of the putative second messenger inositol trisphosphate. The time course, concentration dependence, and P2-purinergic receptor specificity were similar for ATP-stimulated prostacyclin formation and ATP-stimulated inositol phospholipid metabolism. Thus, the increase in prostacyclin formation may be secondary to mobilization of intracellular Ca2+ by inositol trisphosphate, leading to activation of phospholipase A2, liberation of arachidonic acid, and the conversion of arachidonic acid to prostacyclin. We propose that the function of ATP, which is often colocalized with cell-specific hormones in secretory cells, may be to regulate blood flow in the adrenal medulla and other endocrine tissues by interacting with adjacent endothelial cells. PMID- 3039500 TI - Expression and nuclear envelope localization of biologically active fusion glycoprotein gB of herpes simplex virus in mammalian cells using cloned DNA. AB - Herpes simplex virus (HSV) is known to bud from the inner membrane of the nuclear envelope. The structural gene for the glycoprotein gB, which is essential for virus entry and cell fusion induced by HSV type 1, has been cloned in a transient expression vector containing the adenovirus major late promoter, tripartite leader sequence, and VARNA (a special RNA in adenovirus-infected cells) genes. Synthesis and glycosylation of glycoprotein gB was observed in COS-1 cells transfected with the vector containing the gB gene. Removal of a 3' fragment of the cloned gene resulted in the synthesis and secretion of a truncated gB glycoprotein. Immunofluorescence studies revealed that the expressed glycoprotein was localized in the nuclear envelope as well as in the cell surface. The expressed gB-1 glycoprotein was biologically active and induced fusion of cells to produce polykaryons. These data show that HSV glycoprotein gB expressed from cloned gene can be used as a model to study targeting of proteins into the nuclear envelope as well as cell fusion induced by the virus. PMID- 3039499 TI - Molecular cloning and analysis of functional cDNA and genomic clones encoding bovine cellular retinoic acid-binding protein. AB - A recombinant cDNA clone, pCRABP-HS1, encoding cellular retinoic acid-binding protein was isolated from a bovine adrenal cDNA library. COS-7 cells transfected with pCRABP-HS1 produced a biologically active retinoic acid-binding protein molecule of the expected molecular mass (15.5 kDa). RNA blot hybridization analysis using pCRABP-HS1 as a probe revealed a single 1050-nucleotide mRNA species in bovine adrenal, uterus, and testis, tissues that contain the highest levels of retinoic acid-binding activity. No hybridization was detected in RNA extracted from ovary, spleen, kidney, or liver, which contain relatively low levels of cellular retinoic acid-binding protein activity. Analysis of genomic clones isolated from an EcoRI bovine genomic library demonstrated that the bovine cellular retinoic acid-binding protein gene is composed of four exons and three introns. Two putative promoter sequences were identified in the cloned 5' sequence of the gene. PMID- 3039501 TI - Alternative splicing of human elastin mRNA indicated by sequence analysis of cloned genomic and complementary DNA. AB - Poly(A)+ RNA, isolated from a single 7-mo fetal human aorta, was used to synthesize cDNA by the RNase H method, and the cDNA was inserted into lambda gt10. Recombinant phage containing elastin sequences were identified by hybridization with cloned, exon-containing fragments of the human elastin gene. Three clones containing inserts of 3.3, 2.7, and 2.3 kilobases were selected for further analysis. Three overlapping clones containing 17.8 kilobases of the human elastin gene were also isolated from genomic libraries. Complete sequence analysis of the six clones demonstrated that: the cDNA encompassed the entire translated portion of the mRNA encoding 786 amino acids, including several unusual hydrophilic amino acid sequences not previously identified in porcine tropoelastin, exons encoding either hydrophobic or crosslinking domains in the protein alternated in the gene, and a great abundance of Alu repetitive sequences occurred throughout the introns. The data also indicated substantial alternative splicing of the mRNA. These results suggest the potential for significant variation in the precise molecular structure of the elastic fiber in the human population. PMID- 3039502 TI - Effects of 8-bromo-cGMP on Ca2+ levels in vascular smooth muscle cells: possible regulation of Ca2+-ATPase by cGMP-dependent protein kinase. AB - The effects of 8-bromo-cGMP on intracellular calcium concentrations in cultured rat aortic smooth muscle cells were studied. Both angiotensin II and depolarizing concentrations of K+ stimulated Ca2+ accumulation in the cytoplasm. The increase in Ca2+ due to angiotensin II was associated with an increase in inositol phosphates, while that due to K+ was not. Preincubation of cells with 8-bromo cGMP (100 microM) caused an inhibition of peak Ca2+ accumulation to either angiotensin II or K+. To probe the mechanism of action of cGMP in vascular smooth muscle, the effects of cGMP-dependent protein kinase on Ca2+-ATPase from the cultured cell particulate material were investigated. Ca2+-activated ATPase was stimulated approximately equal to 2-fold by exogenous calmodulin and up to 4-fold by low concentrations of purified cGMP-dependent protein kinase. The inclusion of both calmodulin and cGMP-dependent protein kinase resulted in an additive stimulation of Ca2+-ATPase. Stimulation of Ca2+-ATPase activity was observed at all Ca2+ concentrations tested (0.01-1.0 microM). cAMP-dependent protein kinase catalytic subunit and protein kinase C were either ineffective or less effective than cGMP-dependent protein kinase in stimulating the Ca2+-ATPase from rat aortic smooth muscle cells. These results suggest a possible mechanism of action for cGMP in mediating decreases in cytosolic Ca2+ through activation of a Ca2+-ATPase and the subsequent removal of Ca2+ from the cell. PMID- 3039503 TI - Activation of transforming potential of the human insulin receptor gene. AB - A retrovirus containing part of the human insulin receptor (hIR) gene was constructed by replacing ros sequences in the avian sarcoma virus UR2 with hIR cDNA sequences coding for 46 amino acids of the extracellular domain and the entire transmembrane and cytoplasmic domains of the beta subunit of hIR. The resulting virus, named UIR, contains the hIR sequence fused to the 5' portion of the UR2 gag gene coding for p19. UIR is capable of transforming chicken embryo fibroblasts and promoting formation of colonies in soft agar; however, it does not form tumors in vivo. A variant that arose from the parental UIR is capable of efficiently inducing sarcomas in vivo. UIR-transformed cells exhibit higher rates of glucose uptake and growth than normal cells. The 4-kilobase UIR genome codes for a membrane-associated, glycosylated gag-hIR fusion protein of 75 kDa designated P75gag-hir. P75gag-hir contains a protein tyrosine kinase activity that is capable of undergoing autophosphorylation and of phosphorylating foreign substrates in vitro; it is phosphorylated at both serine and tyrosine residues in vivo. PMID- 3039504 TI - Construction of a Leydig cell line synthesizing testosterone under gonadotropin stimulation: a complex endocrine function immortalized by cell hybridization. AB - Hybridization between a mouse Leydig tumor cell line, MA-10, which produces cyclic AMP and progesterone under human chorionic gonadotropin (hCG) stimulation, and freshly isolated mouse Leydig cells gave rise to 54 hybrid clones, one of which, LK17, was capable of hCG-stimulated testosterone production. Subcloning of this hybrid resulted in the emergence of a subclone, K9, whose testosterone production is more than 10 times that of parent clone LK17, after hCG stimulation, with an ED50 of 37 pM. Testosterone synthesis by K9 cells was multiplied by 25 after gonadotropin stimulation, and binding of hCG declined after prolonged exposure to the hormone. These similarities with murine Leydig cells in primary culture make the K9 clone an attractive alternative for physiological studies. PMID- 3039505 TI - An internalized transmembrane protein resides in a fusion-competent endosome for less than 5 minutes. AB - We have used our assay for the cell-free reconstitution of vesicle fusion occurring in endocytosis to investigate the fusion competence of defined endosomal fractions containing the G protein of vesicular stomatitis virus, G protein was first implanted into the plasma membrane, and endocytosis was then allowed to proceed for defined periods of time. Endosomal fractions were prepared by "immuno-isolation" on a solid support with a monoclonal antibody against the cytoplasmic domain of the G protein. Maximal internalization of the G protein occurred within 5 min at 37 degrees C. From this early endosome the G molecules follow a branched pathway: 50% recycles to the cell surface, while 50% is transported along the endocytic route to the lysosomal compartment. The proportion of G protein following each pathway can be modulated. When the amount of implanted G protein was increased, the fraction of G molecules recycling to the cell surface was reduced. When the G molecules were cross-linked with an antibody prior to the internalization step, recycling to the cell surface was abolished. The cell-free analysis of vesicle fusion was carried out with endosomal fractions immuno-isolated after 5, 10, 15, and 30 min of G-protein internalization at 37 degrees C. Fusion competence was at a maximum with the fraction isolated 5 min after internalization and then decreased with a half-life of approximately equal to 3 min with fractions isolated at later time points. The fusion-competent compartment is the early endosome where sorting of the transmembrane G protein to recycling or degradation occurs. PMID- 3039506 TI - Synthesis and secretion of platelet-derived growth factor by human breast cancer cell lines. AB - We report that human breast cancer cells secrete a growth factor that is biologically and immunologically similar to platelet-derived growth factor (PDGF). Serum-free medium conditioned by estrogen-independent MDA-MB-231 or estrogen-dependent MCF-7 cells contains a mitogenic or "competence" activity that is capable of inducing incorporation of [3H]thymidine into quiescent Swiss 3T3 cells in the presence of platelet-poor plasma. In addition, the conditioned medium contains an activity that competes with 125I-labeled PDGF for binding to PDGF receptors on normal human fibroblasts. The secretion of PDGF-like activity by the hormone-responsive cell line MCF-7 is stimulated by 17 beta-estradiol. Like authentic PDGF, the PDGF-like activity produced by breast cancer cells is stable after acid and heat treatment (95 degrees C) and inhibited by reducing agents. The mitogenic activity comigrates with a material of approximately equal to 30 kDa on NaDodSO4/polyacrylamide gels. Immunoprecipitation with PDGF antiserum of proteins from metabolically labeled cell lysates and conditioned medium followed by analysis on nonreducing NaDodSO4/polyacrylamide gels identified proteins of 30 and 34 kDa. Upon reduction, the 30- and 34-kDa bands were converted to 15- and 16-kDa bands suggesting that the immunoprecipitated proteins were made up of two disulfide-linked polypeptides similar to PDGF. Hybridization studies with cDNA probes for the A chain of PDGF and the B chain of PDGF/SIS identified transcripts for both PDGF chains in the MCF-7 and MDA-MB-231 cells. The data summarized above provide conclusive evidence for the synthesis and hormonally regulated secretion of a PDGF-like mitogen by breast carcinoma cells. Production of a PDGF-like growth factor by breast cancer cell lines may be important in mediating paracrine stimulation of tumor growth. PMID- 3039507 TI - Ca2+-mobilizing actions of platelet-derived growth factor differ from those of bombesin and vasopressin in Swiss 3T3 mouse cells. AB - Addition of the mitogenic peptides bombesin and vasopressin to quiescent Swiss 3T3 mouse cells increased the cytosolic Ca2+ concentration without any measurable delay. In contrast, there was a significant lag period (16 +/- 1.2 s) before platelet-derived growth factor (PDGF) increased cytosolic Ca2+ concentration. This lag was not diminished at high concentrations of either porcine or human PDGF. Similar results were obtained in 3T3 cells loaded with quin-2 or fura-2. The differences in the effects of bombesin, vasopressin, and PDGF on Ca2+ movements were also substantiated by measurements of 45Ca2+ efflux and of cellular 45Ca2+ content. Activation of protein kinase C by phorbol esters inhibited Ca2+ mobilization induced by either bombesin or vasopressin. In contrast, phorbol esters had no effect on PDGF-induced cytosolic Ca2+ concentration increase or acceleration of 45Ca2+ efflux. Finally, bombesin and vasopressin caused a rapid increase in the production of inositol 1,4,5 trisphosphate and inositol 1,3,4-trisphosphate, whereas PDGF, even at a saturating concentration, exerted only a small effect. These results indicate that the signal transduction pathways activated by PDGF that lead to Ca2+ mobilization can be distinguished from those utilized by bombesin and vasopressin. PMID- 3039508 TI - Vasoactive intestinal peptide enhances aromatase activity in the neonatal rat ovary before development of primary follicles or responsiveness to follicle stimulating hormone. AB - We have investigated the factors that regulate aromatase activity in fetal neonatal rat ovaries. Ovarian aromatase activity (assessed by measuring the amount of 3H2O) formed from [1 beta-3H]testosterone) is low prior to birth (less than 0.5 pmol/hr per mg of protein) and increases to values greater than 30 pmol/hr per mg of protein between days 8 and 12 after birth. The appearance of ovarian aromatase (postnatal days 2-4) coincides with the development of primordial follicles. Fetal-neonatal ovaries maintained in serum-free organ culture do not develop aromatase activity at the expected time. Ovine follicle stimulating hormone (0.1-1 microgram/ml), ovine luteinizing hormone (0.1 microgram/ml), or their combination failed to induce the enzyme activity in cultured fetal ovaries, whereas follicle-stimulating hormone is effective in preventing the decline in aromatase activity when postnatal day 8 ovaries are placed in culture. In contrast to follicle-stimulating hormone, dibutyryl-cAMP markedly enhances ovarian aromatase in cultured fetal ovaries. Likewise, enhancement of endogenous cAMP formation with forskolin or cholera toxin caused an increase in enzyme activity within 24 hr. Vasoactive intestinal peptide, a peptide known to occur in ovarian nerves, caused a dose-dependent increase in aromatase activity in fetal ovaries prior to folliculogenesis. Of related peptides tested, only the peptide having N-terminal histidine and C-terminal isoleucine amide was capable of inducing aromatase activity in fetal ovaries. The fact that VIP can induce aromatase activity in fetal rat ovaries prior to follicle formation and prior to responsiveness to follicle-stimulating hormone suggests that this neuropeptide may play a critical role in ovarian differentiation. PMID- 3039509 TI - Major histocompatibility complex gene organization in the mole rat Spalax ehrenbergi: evidence for transfer of function between class II genes. AB - A genomic DNA library prepared from the kidney of the mole rat Spalax ehrenbergi was screened with mouse probes representing major histocompatibility complex genes that encode alpha and beta polypeptide chains of class II molecules (alpha and beta genes). Restriction maps were constructed for the cross-hybridizing clones, and the class II genes borne by these clones were identified. By this procedure, five main regions containing class II genes were established. One region contained four genes and two gene fragments, the second region contained two genes, the third region contained one gene and one gene fragment, and the remaining two regions contained one gene each. Altogether, six beta genes, two alpha genes, and three alpha-gene fragments were identified. Two of the genes (one alpha and one beta) were established as belonging to the DQ subclass, and all other genes were found to be members of the DP subclass. (Subclass designations are based on the human HLA class II genes). No genes belonging to the DR and DO (DZ) subclasses were found in the library. The absence of DR genes in S. ehrenbergi was also indicated when other experimental methods were used. At least some of the DP loci are polymorphic and most likely also functional. Thus, in the evolution of the mole rat, the DR (and probably also the DO) loci have been deleted and their function(s) has been taken over by the DP loci, which have expanded to a great extent. These findings argue for functional interchangeability of the individual subclasses of class II loci. PMID- 3039510 TI - Amplification and deregulation of MYC following Epstein-Barr virus infection of a human B-cell line. AB - In Epstein-Barr virus (EBV)-positive Burkitt lymphoma (BL) the role of EBV in the translocation and deregulation of the MYC oncogene remains unknown. By utilizing an EBV-negative BL (BJAB) and several EBV-positive sublines derived from it by in vitro infection, it was possible to show that the presence of the virus was associated with altered expression and copy number of MYC. In the EBV-negative BJAB line, the level of MYC transcripts declined progressively as cells approached the stationary phase of growth. In contrast, in EBV-infected BJAB cells MYC expression remained elevated as cells entered stationary phase. This effect on MYC expression was reversibly linked to the presence of the virus. Furthermore, following EBV infection of BJAB cells by two different strains of EBV, amplification of MYC in association with the appearance of a homogeneously staining region on chromosome 8 at the mapped location of MYC had occurred. These studies suggest that both the deregulation of MYC transcription and the chromosomal rearrangement in the region of the MYC locus in this B-cell line may have occurred as a result of EBV infection. PMID- 3039511 TI - Complete nucleotide sequence of a gene conferring polymyxin B resistance on yeast: similarity of the predicted polypeptide to protein kinases. AB - Polymyxin B is an antibiotic that kills sensitive cells by disrupting their membranes. We have cloned a wild-type yeast gene that, when present on a high copy-number plasmid, renders the cells resistant to the drug. The nucleotide sequence of this gene is presented. A single open reading frame within the sequence has the potential to encode a polypeptide (molecular mass of 77.5 kDa) that shows strong homologies to polypeptides of the protein kinase family. The gene, PBS2, located on chromosome X, is not allelic to the previously described PBS1 gene (where PBS signifies polymyxin B sensitivity). Although pbs1 mutations confer resistance to high levels of polymyxin B, double mutants, pbs1 pbs2, are not resistant to the drug, indicating that PBS2 is essential for pbs1 activity. Models based on the proposed protein kinase activity of the PBS2 gene product are presented to explain the interaction between PBS1 and PBS2 gene products involved in conferring polymyxin B resistance on yeast cells. PMID- 3039512 TI - RNA splicing permits expression of a maize gene with a defective Suppressor mutator transposable element insertion in an exon. AB - The bz-m13CS9 allele of the bronze-1 gene in maize contains a 902-base-pair defective Suppressor-mutator (dSpm) transposable element in the second exon. Nevertheless, 40-50% of the enzymatic activity conditioned by a nonmutant allele at the bronze-1 locus is routinely recovered in crude extracts prepared from plants carrying bz-m13CS9 in the absence of an autonomous Suppressor-mutator element. Analyses of RNAs produced by such plants show that transcription proceeds through the dSpm. The dSpm sequence of the messenger RNA precursor is then removed by RNA splicing using the donor site of the single bronze-1 intron and an acceptor site within the inverted terminal repeat of the dSpm. This results in a messenger RNA with the proper reading frame that could produce a functional enzyme. These data demonstrate that this dSpm insertion in an exon of a structural gene has produced a functional allele with a novel intron consisting, in part, of the dSpm. This mechanism appears to allow dSpm elements to reduce the impact of their insertions on gene expression. PMID- 3039513 TI - Role of antibodies to murine leukemia virus p15E transmembrane protein in immunotherapy against AKR leukemia: a model for studies in human acquired immunodeficiency syndrome. AB - Previous studies have demonstrated that the onset of AKR leukemia could be dramatically delayed and the overall incidence significantly reduced following treatment with high-titered heterologous antibodies directed against the gp71 major glycoprotein of the virus. However, to be maximally successful, the treatment had to be initiated during the postnatal period of the AKR mouse, encompassing a narrow window representing approximately the first 3 days of life. In the present study we sought to extend this barrier by including antibodies directed against a second envelope component of the virion, the transmembrane protein, p15E. We demonstrate that although neither antibodies to gp71 nor antibodies to p15E could influence the course of leukemia development when applied individually later in life, a combination of the two antibodies was effective even if given as late as 5 months after birth. The significance of these studies is discussed in relation to human retrovirus-associated diseases. PMID- 3039514 TI - Endogenous leukotriene D4 formation during anaphylactic shock in the guinea pig. AB - Experiments on the metabolism and excretion of i.v. administered selectively labeled [3H8]leukotriene C4 in bile duct-cannulated guinea pigs indicated predominantly biliary excretion of tritium. The major leukotriene metabolite in bile was identified as leukotriene D4. By monitoring leukotriene excretion radioimmunochromatographically, it was shown that guinea pigs suffering from anaphylactic shock produce leukotriene D4 endogenously. Immunological challenge of animals sensitized to ovalbumin was accompanied by an increase of biliary leukotriene D4 concentrations from 10 +/- 1 to 86 +/- 10 nM (mean +/- SEM, n = 5, P less than 0.001). When considering that bile flow was decreased to about half after challenge, the excretion rate of leukotriene D4 in bile increased from 0.88 +/- 0.16 before to 3.18 +/- 0.38 pmol X min-1 X kg-1 after challenge (mean +/- SEM, n = 5, P less than 0.002). It is concluded that systemic anaphylaxis in the guinea pig is associated with endogenous generation of leukotriene C4 (up to 1 nmol/kg during a 30-min period after the challenge. PMID- 3039516 TI - Morphological changes of an inflammatory myopathy in rhesus monkeys with simian acquired immunodeficiency syndrome. AB - Eleven of 25 rhesus monkeys which died of simian acquired immunodeficiency syndrome (SAIDS) caused by infection with a type D retrovirus related to Mason Pfizer monkey virus showed evidence of muscle weakness and atrophy and had elevated levels of muscle enzymes. Biopsies of affected muscle studied with enzyme histochemistry showed the characteristic features of polymyositis. Inflammatory cells consisting of lymphocytes, macrophages, and large vacuolated bizarre-shaped cells of undetermined type were surrounding or invading muscle fibers and were present in the perivascular spaces and endomysia septa. Within the perivascular infiltrates, lymphocytes were abundant but very few macrophages were present. Other myopathic features including profound proliferation of fibrous tissue, necrosis, and phagocytosis of muscle fibers were noted to a variable degree. The retrovirus was isolated from affected muscles. The clinical and historical features of polymyositis in rhesus monkeys with SAIDS are very similar to those of human polymyositis. The polymyositis in SAIDS induced by a type D retrovirus related to Mason-Pfizer monkey virus is an excellent primate model to study the mechanism and morphological changes of viral-induced muscle damage. PMID- 3039515 TI - Cell transformation and tumor induction by Abelson murine leukemia virus in the absence of helper virus. AB - We investigated the role of the Moloney helper virus, Moloney murine leukemia virus (Mo-MuLV), in cell transformation and tumor induction by the defective Abelson murine leukemia virus (Ab-MuLV). A molecular clone of Ab-MuLV (P160 strain) was transfected into the psi 2 packaging cell line, and helper virus-free Ab-MuLV (psi 2) was harvested from the supernatant medium. Ab-MuLV (psi 2) was as efficient as helper virus-containing Ab-MuLV (Mo-MuLV) in the transformation of primary bone marrow cells in vitro. Inoculation of weanling BALB/c mice with Ab MuLV (psi 2) induced nonthymic pre-B-cell lymphomas with high efficiency and short latency (28 days). Adult BALB/c mice were less sensitive to tumor induction by a factor of 100. Ab-MuLV (psi 2) did not induce tumors in weanling C57BL/6 mice, unlike Ab-MuLV (Mo-MuLV). Examination of the proviral integration pattern in Ab-MuLV (psi 2)-induced tumor cell DNA revealed that each of the tumors contained a single integrated provirus. Immunoprecipitation of viral-encoded proteins in helper virus-free tumor cell lines detected the P160 Ab-MuLV transforming protein; however, no trace of the gag, pol, and env helper virus encoded proteins was found. Our results indicate that integration and expression of a single Ab-MuLV genome is sufficient for efficient transformation of primary bone marrow cells by Ab-MuLV in vitro and tumor induction in susceptible mice. However, the helper virus may contribute to tumor induction in weanling resistant mice. PMID- 3039517 TI - Histamine receptor effects on dissipation of an intracellular proton gradient of isolated gastric mucosal surface cells. AB - The effects of histamine and several H1 and H2 receptor agents on Na+/H+ and Cl /HCO-3 exchange systems of isolated gastric mucosal surface cells were studied. The cells were acid-loaded by the NH4Cl prepulse technique and the spontaneous Na+- and HCO-3-induced dissipation of the intracellular proton gradient (pHi) was followed using the metachromatic dye acridine orange. Histamine (10(-2-5) M) stimulates HCO-3-induced dissipation of the pHi but has no effect on Na+-induced or spontaneous dissipation. The H1 agonist 2-(2-aminoethyl)pyridine and the H2 agonist dimaprit also have no effect on Na+-induced or spontaneous pHi dissipation. However, both of these agents mimic the effect of histamine on HCO-3 induced dissipation, but only at a higher concentration (10(-3) M). The combination of 2-(2-aminoethyl)pyridine and dimaprit produces a histamine-like effect at lower concentrations (10(-5) and 10(-4) M). The effects of histamine are blocked by either the H1 antagonists diphenhydramine and pyrilamine or the H2 antagonists cimetidine and SKF 93479. The results suggest that the effect of histamine on HCO-3-induced dissipation of a pHi in gastric mucosal surface cells is mediated through a coordinated mechanism involving both H1 and H2 receptor sites. PMID- 3039518 TI - Iron-induced accumulation of hepatic metallothionein: the lack of glucocorticoid involvement. AB - The process(es) by which parenteral iron effects the accumulation of hepatic metallothionein (MT) is not known. The present study examined glucocorticoids as potential mediators of this process. Chicks were given either one injection (ip) of iron (+1FE) at 10 mg Fe/kg, two injections of iron (+2FE) given 24 hr apart, or a single injection of saline. Plasma corticosterone was evaluated at various times following the last injection. Plasma corticosterone increased approximately 50% following +1FE but more than 200% at 2 and 4 hr following a second injection of iron (+2FE). Plasma zinc showed a transient increase followed by a considerable depression. Coincidentally, the accumulation (determined at 24 hr) of zinc MT in liver of +2FE chicks was three times higher than that of +1FE chicks. In another experiment, markedly greater changes, at similar time intervals, in plasma corticosterone were effected by multiple subcutaneous injections of adrenocorticotropic hormone (ACTH) (either 5 IU ACTH or 20 IU ACTH/kg). Subsequent analysis of hepatic zinc MT showed only minor changes as a result of ACTH injections. These results indicate that a change in the plasma glucocorticoid corticosterone is not a primary component in the process(es) by which parenteral iron effects an increase in hepatic zinc MT. PMID- 3039519 TI - Hepatic necrosis induced by norepinephrine in rabbits. AB - Extensive hepatic necrosis was produced in rabbits 48 hr following infusion of a cardiopathogenic dose of norepinephrine (NE, 2 micrograms/kg/min for 90 min). Livers had necrotic areas of varying sizes and gross appearances. Histologically, the lesions were areas of varying sizes and gross appearances. Histologically, the lesions were areas of lytic-coagulative necrosis with massive mineralization by calcium. In addition, the serum glutamate-pyruvate transaminase (GPT) was significantly elevated (P less than 0.001). Pretreatment with the alpha 1 adrenoceptor blocker prazosin (200 micrograms/kg) 15 min prior to the standard NE infusion prevented both liver necrosis and serum GPT elevation. It is concluded that large doses of NE produce tissue injury in the liver. This may be the result of excessive activation of the alpha 1-adrenoceptor system, which leads to hepatic ischemia and necrosis. PMID- 3039520 TI - Herpes simplex virus: recurrent and nonrecurrent strains. AB - A study of three Herpes Simplex strains with different frequencies of recurrent disease was done using the New Zealand white rabbit eye model. Each of the three strains, the McKrae strain (high frequency), the E-43 strain (low, frequency), and the CGA-3 (no recurrence) grew well in the rabbit corneal epithelium and produced overt recognizable disease for up to 5 days post-infection, thus minimizing differences in virus reactivation due to a lack of or insufficient ganglionic colonization. Asymptomatic shedding and spontaneous recurrences, as well as iontophoretically induced recurrences, were seen in the E-43 and McKrae strains, but not in the CGA-3-infected animals. The virus strain's optimum temperature was an important aspect of its reactivation process, as shown by the failure of the nonrecurrent CGA-3 to replicate at the host's core temperature (39 degrees C). The fact that these explants yielded infectious virus at 33 degrees C and not at 39 degrees C confirmed that the CGA-3 had colonized the ganglia, and its lack of recurrences or shedding suggests a temperature-dependency relationship. Our observations were further supported by the preferential growth at 39 degrees C of fresh clinical isolates obtained from HSV encephalitis and herpes labialis. Isolates from animals infected with the heterogeneous McKrae were classified as shedders (isolated in the absence of disease) and recurrent (isolated from a recurrent lesion). Both shedders and recurrent isolates were of a homologous nature and retained their phenotype when tested. From this study, we theorize that reactivation and disease may have different regulatory mechanisms. The type of recurrent disease (lesions, asymptomatic shedding, or none) is virus dependent and frequency of disease may be regulated by host functions. PMID- 3039521 TI - Identification of a cDNA encoding the tumor-associated aldehyde dehydrogenase of rat liver. PMID- 3039522 TI - Studies on the hormonal regulation of rat liver alcohol dehydrogenase. AB - These observations have extended and clarified the earlier reports on the effects of thyroid hormone and androgens on liver ADH activity. Thyroid hormone exerts a direct effect on ADH activity, but is not required for the expression of the enzyme. At present, it has not been possible to demonstrate a physiological role for thyroid hormone in determining the developmental and sexual variation in ADH activity. Androgens probably modulate the pattern of secretion of GH, which then determines the level of ADH expression in the liver. We do not understand how GH exerts an effect on liver protein expression in this or other systems. This effect of growth hormone is probably the major controlling factor in the sexual variation in ADH activity. Since the pattern of GH secretion can be influenced by perinatal factors and by stress, it will be of great interest to see if the expression of any of the human ADH isoenzymes is also modified by the pattern of GH secretion. PMID- 3039523 TI - Oxygen radicals and lipid peroxidation in experimental shock. PMID- 3039524 TI - Oxygen radicals scavenging in prophylaxis and treatment of experimental shock. PMID- 3039525 TI - Leukotrienes as mediators in endotoxin shock and tissue trauma. PMID- 3039526 TI - Generation of leukotrienes in polytraumatic patients with adult respiratory distress syndrome (ARDS). PMID- 3039527 TI - Perturbation of transmembrane signaling mechanisms in acute and chronic endotoxemia. AB - Our results reviewed here may be summarized as follows: 1. Continuous endotoxemia significantly interferes with Ca2+-dependent information flow in the liver. 2. The subcellular sites where these molecular lesions can be localized include: a.) the plasma membrane-there are effects at the level of alpha 1-adrenergic and vasopressin binding, and also in the coupling of receptor activation to inositol lipid metabolism in terms of PIP2 degradation and resynthesis b.) the endoplasmic reticulum in terms of Ca2+ release and PI synthesis. Another one of the sequelae of Ca2+-associated receptor activation, namely, cytosolic ionized Ca2+ concentration is also affected. 3. Finally, in addition to seeing the impact of acute or continuous endotoxemia at the level of receptor activation and signal generation, we can also document alterations in the expression of physiologic function subserved by these Ca2+- and inositol lipid-associated signaling processes--i.e. in glycogen phosphorylase activity-being consistent with the above described changes. In conclusion, we state that a causal link is shown between receptor binding, agonist-induced phosphoinositide hydrolysis, intracellular Ca2+ mobilization and activation of phosphorylase a in the liver, suggesting that these alterations may underlie some of the metabolic consequences of chronic sepsis. PMID- 3039528 TI - Quantification of granulocyte enzymes/proteins with immunoassays. PMID- 3039529 TI - Involvement of oxygen free radicals in the metabolism and toxicity of ethanol. PMID- 3039530 TI - Ethanol tolerance and dependence. PMID- 3039531 TI - Alcohol, acetaldehyde and salsolinol-induced alterations in function and metabolism of cerebral GABAergic and cholinergic neurons: possible involvements in alcohol dependence and withdrawal. PMID- 3039532 TI - Formation of leukotriene B4, 20-hydroxy leukotriene B4 and other arachidonic acid metabolites by macrophages during peritonitis in patients with continuous ambulatory peritoneal dialysis. AB - Macrophages, isolated from dialysis fluid of three patients with continuous ambulatory peritoneal dialysis (CAPD) at different times during peritonitis were labelled with 14C-arachidonic acid and stimulated with the calcium ionophore A23187. The main metabolites formed by 5-lipoxygenase activity were leukotriene B4 (LTB4) and 5-hydroxy-6, 9, 11, 14-eicosatetraenoic acid (5-HETE). Smaller amounts of cyclooxygenase metabolites were present and also a major compound with an elution time between 6-keto-prostaglandin F1 alpha (6-keto-PGF1 alpha) and thromboxane B2 (TxB2). This substance was isolated, analyzed by GC-MS and identified as 20-hydroxy-leukotriene B4 (20-OH-LTB4). This indicates that human peritoneal macrophages obtained from CAPD not only produce leukotrienes and prostaglandins, but also the omega-hydroxylase product of LTB4, which has been demonstrated to be present in polymorphonuclear leucocytes. The activity of this enzyme was not correlated with the severity of the peritonitis. PMID- 3039533 TI - Arachidonic acid metabolism by lipoxygenase pathways in uterine and intrauterine tissues of pregnant sheep. AB - The formation of arachidonate lipoxygenase products by uterine and intrauterine tissues of sheep in the last third of gestation has been evaluated. Maternal and fetal cotyledon, myometrium and fetal membrane exhibited evidence of arachidonate 5-, 12-, and 15- lipoxygenase activities. The major lipoxygenase product formed by fetal membrane and fetal cotyledon was leukotriene B4 (LTB4) whereas maternal cotyledon and myometrium produced mainly 12-hydroxyeicosatetraenoic acid (12 HETE). Arachidonate lipoxygenase products may play significant roles in the regulation of fetal and uteroplacental hemodynamics and it is speculated that the formation of leukotriene B4 predominantly by tissues of fetal origin may be of significance in the immunologic adaptations of pregnancy. PMID- 3039534 TI - Metabolism of leukotriene B4 by polymorphonuclear granulocytes of severely burned patients. AB - Leukotriene B4 release from polymorphonuclear granulocytes of severely burned patients was reduced as compared to healthy donor cells. This decrease is due to an enhanced conversion of LTB4 into the 20-hydroxy- and 20-carboxy-metabolites and further to a decreased LTB4-synthesis. In addition, studies on the exogenous LTB4-conversion revealed an unidentified compound which was derived from LTB4. Our data suggest a modulation of the enzymatic activities involved in omega oxidation of LTB4 (isoenzymes of cytochrome P-450). PMID- 3039535 TI - Altered arachidonic acid metabolism in granulocytes of polytraumatized patients. AB - The generation and metabolism of leukotrienes (LTs) from polymorphonuclear granulocytes of four polytraumatic patients on stimulation with the Ca-Ionophore A 23187 were studied by high performance liquid chromatography. In contrast to healthy donors the concentration of LTB4 within the supernatant of the stimulated granulocytes from these patients is reduced. The ratio of LTB4 versus the combined amounts of the biologically inactive 6-trans and 12-epi-6-trans-isomers is significantly decreased. In one patient who suffered from an Adult Respiratory Distress Syndrome (ARDS) a pronounced enhancement of LTC4 synthesis was observed. PMID- 3039536 TI - Antiatherogenic action of eicosapentaenoic acid (EPA) in multiple oral doses. AB - The possible antiatherogenic action of eicosapentaenoic acid (EPA) was pharmacologically investigated using purified and ethylesterified fish oil containing 75% EPA (EPA-E) in multiple oral doses in rats and rabbits. EPA-E showed dose-dependent prevention of thrombus formation in a vascular shunt or sudden death caused by arachidonic acid injection in rats. EPA-E in daily doses ranging from 3 to 30 mg/kg slightly altered platelet aggregability and prostacyclin-like activity generated from arterial ring preparations of rats, but these alterations were not statistically significant. Further, EPA-E showed no effect on blood viscosity of rats. In cholesterol-fed rabbits, EPA-E in daily doses of 10 and 30 mg/kg moderately lowered the levels of plasma cholesterol, beta-lipoprotein, triglyceride and phospholipid, but these changes showed neither dose-dependency nor time-dependency. In this experiment, EPA-E moderately altered atherogenic plaque formation and platelet aggregability, but these alterations were not statistically significant. EPA-E showed no effect on prostacyclin-like activity generated from arterial ring preparations and blood viscosity of cholesterol-fed rabbits. It is, therefore, proposed that the antithrombotic action of EPA-E may be partially related to its effects on platelet aggregability and prostacyclin generation, but the major mechanism remains unclear. PMID- 3039537 TI - Development of a system for evaluating 5-lipoxygenase inhibitors using human whole blood. AB - A reliable system for evaluating 5-lipoxygenase (5-LO) pathway inhibitors employing human whole blood stimulated by the calcium ionophore, A-23187, and yeast cell walls (YCW) is described. In developing this system, we have shown that leukotriene B4 (LTB4) and 5-hydroxyeicosatetraenoic acid (5-HETE) can be recovered quantitatively from whole blood, and can be measured with accuracy and a precision (standard deviation) of +/- 12%. Apparent differences in LTB4/5-HETE levels between donors can be minimized by normalizing the LTB4/5-HETE production to neutrophil number. Variability in LTB4/5-HETE production among different donors was reduced by increasing the ionophore concentration. The kinetics of ionophore stimulated product production display a 1-4 min lag which is dependent on ionophore concentration. The lag is removed by pretreatment of blood with 5 micrograms/ml cytochalasin B. Likewise, the kinetics of product formation after stimulation with yeast cell walls demonstrated a lag period, which could be shortened by prior opsonization of the YCW. The amount of LTB4 metabolism to 20 OH-LTB4 and 20-COOH-LTB4 in this system is approximately 20%. Phenidone, nordihydroguaiaretic acid, and nafazatrom, known inhibitors of the 5-LO pathway, display half-maximal inhibition points of 0.4, 1.5, and 9 micrograms/ml, respectively. In summary, we believe that this assay offers a guide for predicting systemic levels of drug needed to be achieved for effective inhibition of cellular LTB4/5-HETE synthesis/release in humans. PMID- 3039539 TI - Reserpine-induced hypothermia: participation of beta 1 and beta 2 adrenergic receptors. AB - In mice, reserpine-induced hypothermia is partly antagonized by clenbuterol, a beta-adrenergic agonist specific for beta 2 receptors, and completely antagonized by dobutamine, a beta-adrenergic agonist specific for beta 1 receptors. In addition, the effects of dobutamine and of clenbuterol are impaired by betaxolol (1 and 4 mg/kg) and unchanged by ICI 118,551 (1 and 4 mg/kg) beta-blocking drugs respectively selective for beta 1 and beta 2 receptors. These results indicate that reserpine-induced hypothermia depends on the beta 1 receptors and lend support to an indirect effect of clenbuterol. After a chronic treatment, clenbuterol-induced antagonism of reserpine hypothermia is facilitated. This facilitation is impaired by ICI 118,551 and by betaxolol but, in this last case, with high doses only. So the facilitation involves beta 2 adrenergic receptors and implies an increase in the sensitivity of beta 1 receptors. PMID- 3039538 TI - Neonatal exposure to a high level of ACTH4-10 impairs adult learning performance. AB - Male offspring of Sprague-Dawley dams were injected with 25 micrograms of ACTH4 10 or the vehicle on days 2 through 7 postpartum. Peptide treated animals exhibited a marked motor response to the peptide injection. Adrenal weights of these animals were consistently heavier than littermate controls in both the developing and adult animals. ACTH4-10 treated neonates exhibited significantly poorer learning performance in the shuttle box and were slower to acquire the reversal learning problem of a visual discrimination task under light shock levels. In addition, these animals also exhibited an exaggerated startle response and a stronger thigmotaxis response in the open field than controls. These results indicate that exposing the developing nervous system to relatively high levels of ACTH4-10 can produce marked long-term effects on behavior. PMID- 3039540 TI - A benzodiazepine receptor inverse agonist inhibits stress-induced ulcer formation. AB - The effects of a benzodiazepine receptor inverse agonist (FG 7142) on gastric ulcer formation were studied in restrained rats. FG 7142 (10-50 mg/kg) reduced in a dose-dependent fashion both the number and cumulative length of gastric ulcers elicited by restraint for 2 hr at 4 degrees C, but did not affect ulcer formation in unrestrained animals maintained in this environment. FG 7142 also reduced gastric ulcer formation in restrained rats maintained at 22 degrees C for 5 hr. The ability of FG 7142 to reduce restraint-stress induced gastric ulcer formation was blocked by the benzodiazepine receptor antagonist ZK 93426 and the beta adrenoceptor antagonist propranolol. These findings suggest that FG 7142 produces a benzodiazepine-receptor mediated reduction in gastric ulcer formation, which may result from its ability to increase activity of the sympathetic nervous system. PMID- 3039541 TI - [Synergistic effect of lysolecithin analogs on the virostatic action of nucleoside antimetabolites in vitro]. PMID- 3039542 TI - X-ray characterisation of normal and neoplastic breast tissues. AB - Normal and neoplastic breast tissues have been characterised in terms of x-ray attenuation. Samples of normal fat and fibrous tissue were obtained from reduction mammoplasty and autopsy, and infiltrating duct carcinoma specimens from mastectomy and lumpectomy. A high-purity germanium spectroscopy system and a beam of 120 kV constant potential x-rays were used to determine the linear attenuation coefficient from 18 to 110 keV. Densities were determined from buoyancy measurements and were used to obtain mass attenuation coefficients. Infiltrating duct carcinomas and fat are well distinguished by x-ray attenuation. For photon energies used for film-screen mammography, infiltrating duct carcinomas are more attenuating than fibrous tissue. Above 31 keV, the ranges of attenuation of the two tissue types overlap. The attenuation coefficients of tissues have been concisely represented by equivalent thicknesses of lucite and aluminium. Analysis based on the average attenuation properties of tissues indicates that dual-energy mammography, using an ideal imaging system, would require 0.06 cGy to provide images in which 1 cm infiltrating duct carcinomas are displayed with a signal to noise ratio of 5 against a background over which the fat/fibrous contrast has been suppressed. This dose is similar to that currently used in conventional film screen mammography. PMID- 3039543 TI - Synthetic diamonds as ionisation chamber radiation detectors in biological environments. AB - Synthetic diamonds with nitrogen concentrations higher than previously reported in the literature are found to operate very effectively as alpha-particle detectors, as well as detectors for gamma radiation, when operated as ionisation chambers. Certain of the specimens exhibited extensive linear response characteristics when subjected to either alpha particles or gamma radiation of various dose rates. For alpha particles, the response of the detectors at constant particle flux was also found to increase linearly with increasing alpha particle energy. Unlike previously reported investigations, however, the variation in the response of the synthetic stones to gamma radiation as a function of time was found to be not only more rapid but also to be virtually unaffected by illumination with intense white light. PMID- 3039544 TI - [Clinical picture and treatment of the Raynaud syndrome of the upper extremities]. AB - Raynaud's disease occurs in different clinical forms, which are not specific to the vascular spasm which characterizes the disease. This spasm virtually always develops in the same manner. Clinical aspects depend on the underlying pathology, which will define treatment. Therapeutic efficacy in Raynaud's disease is discussed. Current interest focuses on serotonin antagonists. PMID- 3039546 TI - Generation of electronically excited states in situ. Polymorphonuclear leukocytes treated with phenylacetaldehyde. AB - In polymorphonuclear leukocytes phenylacetaldehyde promotes an intracellular O2 consuming process in which myeloperoxidase participates. The reaction is accompanied by lipid peroxidation as shown by both malondialdehyde formation and biphasic light emission. The lipid peroxidation appears to be induced by intracellularly generated triplet benzaldehyde. When chlorophyll-a is solubilized in the leukocytes, biphasic emission is observed in the red, demonstrating that the excited species formed in lipid peroxidation transfer their energy to chlorophylls bound to the cell. The energy transfer process is efficient and does not occur by radiative transfer. PMID- 3039545 TI - [Thromboembolic disease and congenital venous abnormalities]. AB - A pulmonary embolus and a bilateral thrombosis of the internal carotid artery caused the sudden death of two young adults with a congenital venous abnormality. The study of 49 patients suffering from a Klippel-Trenaunay syndrome showed that 11 out of 49 patients, or 22.5 p. cent had thrombosis problems: namely 7 pulmonary emboli and 8 deep thrombophlebitis. Phlebography did not permit to discover any specific anatomical abnormality. 5 out of 11 patients presented avalvular and dilated deep veins. 6 out of 11 presented angioma of the pelvis. In the group of patients with Klippel-Trenaunay who never presented a deep venous thrombosis, the same signs are noted in one third of the cases. It is possible that coagulation insufficiency may explain the high number of thrombophlebitis. However, an extended analysis of 11 patients did not permit to discover any abnormality. But, subsequently, a study of the fibrinolytic activity demonstrated a normal fibrinogen level, while the fibrinopeptide A level was markedly elevated in each case, with an abnormal thrombin activity. PMID- 3039547 TI - Oxygen radicals mediate cell inactivation by acridine dyes, fluorescein, and lucifer yellow CH. AB - Acridine dyes, fluorescein and lucifer yellow CH are fluorescent photosensitizers used experimentally to selectively stain and photodynamically destroy eukaryotic cells and subcellular structures. We have determined that the mechanism of light- and oxygen-dependent inactivation of E. coli by these dyes involves oxygen radicals and hydrogen peroxide. All of the dyes oxidized NAD(P)H+ under illumination. Superoxide (O2), detected as the superoxide dismutase (SOD) inhibitable reduction of ferricytochrome c, was a major product of the dye sensitized photooxidation. Cationic acridine dyes penetrated the membranes of E. coli and were photoreduced intracellularly. Reduced dyes diffused back into the medium and mediated the reduction of extracellular ferricytochrome c. The anionic dyes fluorescein and lucifer yellow CH were unable to mediate extracellular cytochrome c reduction, indicating that these dyes were impermeable to the E. coli membrane. Acridine dyes, when illuminated, inhibited the growth of E. coli in a rich medium, and induced the synthesis of SOD. Fluorescein and lucifer yellow CH did not inhibit growth or induce SOD synthesis because they were unable to enter the cells. Superoxide (O2) and hydrogen peroxide (H2O2), generated by the enzyme xanthine oxidase were toxic to E. coli B. Inactivation by xanthine oxidase was partially inhibited by exogenous SOD and completely inhibited by exogenous catalase or SOD plus catalase. Similarly, exogenous SOD plus catalase protected against inactivation by acridines and fluorescein-NADH or lucifer yellow CH-NADH mixtures. Prior induction of superoxide dismutase and catalase in E. coli B significantly protected cells against a subsequent challenge by illuminated acridine dyes. SOD and catalases preinduction combined with additions of exogenous SOD and catalase completely protected E. coli B against photodynamic inactivation by acridine yellow. The hydroxyl radical scavengers, dimethyl sulfoxide, sodium benzoate and thiourea, protected E. coli B against photodynamic inactivation by acridine orange. The results implicate O2, H2O2, and the hydroxyl radical (OH) as underlying molecular agents of the phototoxicity mediated by acridine orange, acridine yellow, fluorescein and lucifer yellow CH. PMID- 3039548 TI - A portrait of the adenosine triphosphate synthetase-hydrolase. AB - It is proposed that the ATP-synthetic and ATP-hydrolytic activities of energy transducing mitochondria, chloroplasts and bacterial membranes are due to different enzyme systems. It is suggested that the alpha-subunits of the oligomycin-sensitive coupling factor catalyze synthesis and the beta-subunits catalyze hydrolysis. Evidence is assembled from the literature in support of this concept. PMID- 3039549 TI - Survivability and long-term stress reactivity levels following repeated exposure to nuclear magnetic resonance imaging procedures in rats. AB - The effect of exposure to the magnetic and radio-frequency fields associated with Nuclear Magnetic Resonance Imaging (MRI) on survivability and long-term stress reactivity levels was examined in male rats. Rats in the experimental condition were exposed to MRI for 22.5 minutes for five successive days (Expt. 1) or 23.3 minutes for twenty-one successive days (Expt. 2). Sham field exposed animals received the identical treatment as the exposed animals except that the magnetic and radio-frequency fields were absent. Control rats were also maintained. Thirteen to twenty-two months after the exposure procedure the remaining rats were sacrificed and their whole body, spleen, heart, thymus, and adrenal weights were recorded. Blood samples were taken for measurements of red and white blood cell counts, hemoglobin content, as well as adrenocorticotrophin and corticosterone levels. The results fail to provide any evidence for changes in survivability and long-term stress reactivity levels in rats exposed to MRI and thus give no grounds to challenge the view that MRI is a safe diagnostic procedure. PMID- 3039550 TI - ATP-dependent degradation of 125I-bovine serum albumin by rabbit reticulocytes does not need repression of an endogenous inhibitor. AB - The ubiquitin-dependent proteolysis of 125I-bovine serum albumin in rabbit reticulocytes has been investigated. Using various reticulocyte fractions (reticulocyte protease, inhibitor-free protease, "ubiquitin" and "inhibitor") in the presence or absence of ATP, we found that the repression of an endogenous inhibitor, as suggested by others for alpha-casein proteolysis, is unlikely for bovine serum albumin. Therefore, differences exist in the ATP-dependent proteolytic pathway of rabbit reticulocytes depending on the substrate. Fractionation of the reticulocyte ATP-dependent proteolytic system revealed at least two proteolytic and two inhibitory fractions involved in the proteolysis of bovine serum albumin. PMID- 3039551 TI - Body weight gain, food intake and adrenal development in chronic noise stressed rats. AB - Wistar chronic treated rats (30 days) were used to investigate the effect of hypothalamic-pituitary-adrenal activity on growth, food intake and adrenal development (weight and DNA content). The animals were submitted to noise stress, ACTH administration and dexamethasone suppression test. Noise stress decreased body weight gain and food intake. No adrenal hypertrophy was observed but an increase in relative DNA content by stress has been found. ACTH and dexamethasone treated rats showed a body weight and food intake decrease vs. controls. The effect on body weight was higher in dexamethasone treated rats. Adrenal hypertrophy and hyperplasia were found in ACTH treated rats, whereas dexamethasone provoked adrenal atrophy with a decrease in DNA content. PMID- 3039552 TI - Chronic stress depresses exploratory activity and behavioral performance in the forced swimming test without altering ACTH response to a novel acute stressor. AB - The present study investigated the effects of chronic stress on some behavioral and endocrine variables in adult male rats. The variables were always measured the day that followed the stress session to chronically stressed rats. Two stress models were used: chronic shock (CS), and a combination of various stressors randomly chosen each day (CV). Chronic but not acute exposure to the stressors resulted in decreased exploratory activity in the holeboard. Likewise, only chronic exposure to shock or variable stressors resulted in increased immobility in the Porsolt's test. Taken together these results suggest that changes in exploratory and forced swimming activities might be affected by chronic stress. In contrast, neither CS nor CV rats showed altered basal or stress-induced levels of ACTH which suggests that the activity of the axis was unchanged. Although the present data indicate that chronic stress caused behavioral abnormalities closely related to that expected to take place in some type of depressive disorders, endocrine data are not in good agreement with those occurring in depression. PMID- 3039553 TI - Biochemical effects of acute stress on energy metabolism in liver damaged rats. AB - Rats having a non-necrotic damaged liver or a necrotic damaged liver produced by D-galactosamine administration were restrained in water for 0.5, 1, 2, 4, 6, 8 and 12 hours. Serial changes in intrahepatic energy metabolism were compared with those in normal liver. Energy charge, which represents the degree of equilibrium between the energy producing and consuming systems, and cyclic AMP, an intracellular messenger mediating the action of hormones, showed biphasic increases before and after 2 hr in the damaged livers. The lactate/pyruvate ratio, which reflects the cytosolic redox state, markedly increased at 0.5 hr in the damaged livers but returned to the pre-stress level after 1 hr in the non necrotic damaged liver and after 4 hr in the necrotic damaged liver, showing a transient reduced state. The beta-hydroxybutyrate/acetoacetate ratio, which represents the mitochondrial redox state, decreased at 0.5 hr and returned to the pre-stress level at 1 hr in the non-necrotic damaged liver, exhibiting a transient oxidized state. However, in the necrotic damaged liver, the value decreased at 0.5 hr remained low thereafter, demonstrating a persistent oxidized state. These findings show that, in severely damaged liver, stress has more marked effects on hepatic energy metabolism. PMID- 3039554 TI - Effects of intraperitoneal injection of lithium chloride on neurohypophyseal activity: implications for behavioral studies. AB - Intraperitoneal injections of lithium chloride (LiCl) were found to increase the activity of vasopressin-neurons and oxytocin-neurons as indexed by rises in plasma concentrations of vasopressin-associated neurophysin (VP-RNP) and oxytocin associated neurophysin (OT-RNP). Plasma VP-RNP increased 12 and 4 times basal levels (greater than or equal to 20 fmol/ml) reaching values of 248 +/- 37 fmol/ml (3.0 mEq LiCl/kg body weight) and 89 +/- 10 fmol/ml (1.5 mEq LiCl/kg body weight) at 60 minutes. OT-RNP rose to 37-and 10-times basal levels (greater than or equal to 20 fmol/ml) with peak values of 749 +/- 100 fmol/ml and 188 +/- 48 fmol/ml ten minutes following injection of 3.0 or 1.5 mEq LiCl/kg body weight. Mean arterial pressure increased in response to lithium treatment by 31 +/- 6 mm Hg at 60 minutes in rats receiving 3.0 mEq LiCl/kg and by 22.5 +/- 5 mm Hg at 10 minutes in rats receiving 1.5 mEq LiCl/kg over pretreatment values (125 +/- 3 mm Hg). Heart rate decreased from a pretreatment value of 422 +/- 12 beats/min to 367 +/- 48 beats/min at 10 minutes and to 341 +/- 27 beats/min at 20 minutes for rats treated with the high and low dose of lithium, respectively. These findings suggest that the behavioral effects of LiCl could result from multiple mechanisms and involve its acute release of vasopressin and oxytocin. It is also possible that changes in cardiovascular function may act as cues when LiCl is used as an aversive stimulus. PMID- 3039555 TI - Short, interspersed, and repetitive DNA sequences in Spiroplasma species. AB - Small fragments of DNA from an 8-kbp plasmid, pRA1, from a plant pathogenic strain of Spiroplasma citri were shown previously to be present in the chromosomal DNA of at least two species of Spiroplasma. We describe here the shot gun cloning of chromosomal DNA from S. citri Maroc and the identification of two distinct sequences exhibiting homology to pRA1. Further subcloning experiments provided specific molecular probes for the identification of these two sequences in chromosomal DNA from three distinct plant pathogenic species of Spiroplasma. The results of Southern blot hybridization indicated that each of the pRA1 associated sequences is present as multiple copies in short, dispersed, and repetitive sequences in the chromosomes of these three strains. None of the sequences was detectable in chromosomal DNA from an additional nine Spiroplasma strains examined. PMID- 3039556 TI - Characterization of the maintenance functions of IncFIV plasmid R124. AB - The genetic arrangement of the regions involved in R124 replication and incompatibility have been located and their homology to the IncFI basic replicons has been assessed. We show that R124 has homology with all three basic replicons, RepFIA, RepFIB, and RepFIC, and that these regions, FIVA, RepFIVB, and RepFIVC, are widely separated on the R124 genome. Cloning of autonomously replicating fragments has shown that RepFIVB and RepFIVC are functional in R124 and express incompatibility. The FIVA region was unable to form a functional replicon and when cloned into pUC8 lacked incompatibility activity. A fourth region of R124 was identified, which although not essential for replication stabilized mini-R124 plasmid replication and exhibited incompatibility with R124. This region, designated IncIV, showed no homology to RepFIA, RepFIB, or RepFIC. Incompatibility expression of IncIV required only the EcoRI fragment E13 but the strength of the reaction was modified in the presence of other fragments. The replication and incompatibility properties of an R124 deletion derivative indicated that R124 can switch its replication to either RepFIVB or RepFIVC when in the presence of an incompatible plasmid. The ambiguous incompatibility reactions reported for R124 is a result of the expression of the two functional replicons, RepFIVB and RepFIVC, and that expressed by IncIV. PMID- 3039557 TI - Genetic basis of a Tn7 insertion mutation in the trfA region of the promiscuous IncP-1 plasmid R18 which affects its host range. PMID- 3039558 TI - Extrachromosomal systems and gene transmission in anaerobic bacteria. AB - Obligately anaerobic bacteria are important in terms of their role as medical pathogens as well as their degradative capacities in a variety of natural ecosystems. Two major anaerobic genera, Bacteroides and Clostridium, are examined in this review. Plasmid elements in both genera are reviewed within the context of conjugal transfer and drug resistance. Genetic systems that facilitate the study of these anaerobic bacteria have emerged during the past several years. In large part, these developments have been linked to work centered on extrachromosomal genetic systems in these organisms. Conjugal transfer of antibiotic resistance has been a central focus in this regard. Transposable genetic elements in the Bacteroides are discussed and the evolution and spread of resistance to lincosamide antibiotics are considered at the molecular level. Recombinant DNA systems that employ shuttle vectors which are mobilized by conjugative plasmids have been developed for use in Bacteroides and Clostridium. The application of transmission and recombinant DNA genetic systems to study these anaerobes is under way and is likely to lead to an increased understanding of this important group of procaryotes. PMID- 3039559 TI - [Distribution of Pick bodies in the central nervous system of Pick's disease with special reference to their association with neuronal loss]. PMID- 3039560 TI - Nucleotide sequence analysis of cellular flanking regions of intracisternal A particle gene 81. AB - The cellular nucleotide sequences flanking the mouse intracisternal A-particle gene 81 were determined. The results indicated that they were made of many small oligonucleotide repeats both direct and indirect in orientation. These two different kinds of repeating sequences were often found to be overlap. The overall base composition of this region is relatively A + T rich. The most important feature of the sequences determined was that it consists of several repeated dinucleotide tracts containing a (CA)16 repeating cluster in the 5' end flanking region of one strand and another repeating dinucleotide cluster, (GT)16, in the 3' end flanking region of the same strand of this gene. In addition, the existence of two clusters of 9 continuous 5-bp repeat units, GCTTT, was found in the 3' end flanking region. The possible roles of such repeating sequence were discussed. PMID- 3039561 TI - Neural dynamics of decision making under risk: affective balance and cognitive emotional interactions. PMID- 3039562 TI - Lack of effect of bromocriptine and domperidone on arginine-vasopressin response to insulin-induced hypoglycemia in normal men. AB - In order to establish whether the arginine-vasopressin (AVP) increase evoked by insulin-induced hypoglycemia is mediated by a dopaminergic pathway, 16 normal men were given an insulin tolerance test (ITT) under basal conditions and after treatment with the dopamine agonist bromocriptine (2.5 mg by mouth) (eight subjects) or the dopamine antagonist domperidone (10 mg i.v.) (eight subjects). In addition, five out of eight subjects in each group were treated with higher doses of bromocriptine (5 mg by mouth) or domperidone (20 mg i.v.). Basal and ITT stimulated AVP secretion was not modified by the drug treatments, suggesting that dopamine is not involved in the regulation of AVP secretion in response to insulin-induced hypoglycemia. PMID- 3039563 TI - [Problems and coping with problems by tumor patients in inpatient after care]. PMID- 3039564 TI - Effects of a health promotion advertising campaign on sales of ready-to-eat cereals. AB - The objective of this study was to determine how the sales of various segments of the high fiber and nonhigh fiber, ready-to-eat (RTE) cereal market were influenced by a health message advertising campaign about the possible benefits of a high fiber, low fat diet for preventing some types of cancer. The fiber statements in the media campaign were endorsed by the National Cancer Institute (NCI). The campaign was undertaken by the Kellogg Company to promote its line of high fiber cereal products, including Kellogg's All-Bran. The data base consisted of computerized purchase data from 209 Giant Food, Inc., supermarkets in the Baltimore, MD, and Washington, DC, metropolitan areas. All the RTE cereal products in the stores were classified according to their fiber content and competitive market positions compared with Kellogg high fiber cereals. Estimates of market share for the various classes of RTE cereal products were obtained weekly for each store during a period of 64 weeks, beginning 16 weeks before the start of the campaign. Shifts in market share between high fiber and nonhigh fiber cereal classifications indicate substantial increases in consumer purchases of Kellogg high fiber cereals, particularly All-Bran, beginning with the start of the Kellogg advertising campaign. Growth in market share of high fiber cereals continued during the entire 48-week evaluation period, with much of the later growth in non-Kellogg high fiber cereals. Growth in sales of high fiber cereals was mainly at the expense of low fiber cereals such as granola-type products. The implications of these results for the competitive and educational effectiveness of commercially sponsored health and diet messages are discussed. PMID- 3039565 TI - Discussing possible standards of natural radioactivity in building materials. AB - This paper discusses different possibilities of deriving reference values for the natural radioactivity concentrations in building materials to estimate possible additional radiation exposure for the population. Based on comprehensive experimental and theoretical investigations the consequences of the resulting hypothetical reference activity concentrations in building materials, applying different dose limits, were examined. The calculation of the activity concentration standards was performed for standard conditions obtained by earlier studies on exhalation of Radon-222 and Radon-220 from building materials. PMID- 3039566 TI - [Biochemical reception and ionizing irradiation of the body]. AB - The authors present the idea of the importance of studying the mechanism of biochemical reception of biologically active compounds (BAC) particularly the reception of prostaglandins after the effect of ionizing radiation on eucaryotes as a factor playing a significant role in understanding the radiation sickness pathogenesis. The perspectives of studying BAC reception are prognosticated for searching new radiomodifying agents (radioprotectors and means for treating radiation sickness). PMID- 3039567 TI - [Molecular-cellular mechanisms of the biological action of low x-ray doses on isolated mammalian cells]. AB - The colony-formation was stimulated by X-irradiation (0.03-0.30 Gy) of cultured Chinese hamster cells. The stimulation was only noted in the nonadhered cells which was perhaps associated with a change in their adhesive properties. It was shown that low-level radiation induced changes in the structural organization of the cell membrane. PMID- 3039568 TI - [Biological effectiveness of helium ions and relativistic-energy protons]. AB - It was shown that RBE coefficients of protons (9 GeV) and accelerated helium ions (4 GeV/nucleon) are within the range from 1.0 to 11.6 and 1.0 to 7.2, respectively, depending on the object under study, the criterium of estimation, the registration time, and the dose value. PMID- 3039569 TI - [Effect of metronidazole and local irradiation of the tumor on paramagnetic metal complexes of the liver and tumor tissues]. AB - A study was made of changes in the intensity of the ESR signals in tissues of sarcoma-37 and liver of mice after radiation of the tumor, administration of metronidazole, and after the combined effect of the two factors. The most pronounced changes in the ESR signals were induced by metronidazole. An appreciable increase in the content of nitrosyl complexes in the tumor was noted after the combined effect of metronidazole and radiation which was indicative of the radiation-induced formation of a large number of metronidazole anion-radicals in the tumor. PMID- 3039570 TI - Focal liver masses: differential diagnosis with pulsed Doppler US. AB - Duplex Doppler ultrasound (US) was used in 68 consecutive patients with focal liver lesions, including 12 hepatocellular carcinomas, one cholangiocarcinoma, 37 metastases, 15 hemangiomas, one hemangioendothelioma, and two focal nodular hyperplasias. Of the hepatocellular carcinomas, six were diffusely hyperechoic, two were hypoechoic, two were single hyperechoic lesions, and two were multifocal and hyperechoic. All ten tumors with Doppler shifts of 5 kHz or above proved to be hepatocellular carcinomas. The other two hepatocellular carcinomas showed Doppler shifts of 3 kHz. In contrast, no hemangioma showed shifts above 0.7 kHz, and ten of the 15 gave no detectable signal. Of the metastases, 20 gave no signal and 17 had signals of up to 4 kHz. Three-kilohertz signals were also obtained from a cholangiocarcinoma, a hemangioendothelioma, and focal nodular hyperplasia. Correlation with angiographic findings suggested that the high-velocity Doppler signals were associated with large pressure gradients due to arteriovenous shunting. Duplex Doppler US can therefore aid in the differential diagnosis of diffuse and focal liver lesions. PMID- 3039571 TI - Tumors of the parapharyngeal space and upper neck: MR imaging characteristics. AB - Magnetic resonance (MR) imaging characteristics of 40 tumors involving the parapharyngeal space and the upper part of the neck were reviewed. These lesions could be classified as being either hypervascular (glomus tumors or metastatic kidney, thyroid, or venous hemangiomas) or hypovascular (salivary gland tumors, neurogenic tumors, lymphomas, sarcomas). Detailed analysis of the contour of the neoplasm combined with clinical findings allowed further refinement of the differential diagnosis in each category. Most lesions had an intermediate signal intensity on T1-weighted images and a fairly high signal intensity on T2-weighted images. Hypervascular tumors had a number of "channel voids" caused by high-flow vessels on T1- and T2-weighted images, and on T2-weighted images there were areas of high signal intensity, presumably due to sites of slow flow within the image plane. The hypovascular lesions were quite homogeneous, and it was therefore more difficult to differentiate among the neoplasms in this group. PMID- 3039572 TI - The cell biology and development of vasopressinergic and oxytocinergic neurons. PMID- 3039573 TI - Brain vasopressin: from electrophysiological effects to neurophysiological function. PMID- 3039574 TI - Neurohypophyseal hormone receptor systems in brain and periphery. PMID- 3039575 TI - Analysis of receptor-coupled events in neuropeptide action using clonal cell lines. PMID- 3039576 TI - Tachykinin receptors in the CNS. PMID- 3039577 TI - Molecular diversity and cellular functions of neuropeptides. PMID- 3039578 TI - Adaptation and brain function. PMID- 3039579 TI - ACTH neuropeptide stimulation of serotonergic neuronal maturation in tissue culture: modulation by hippocampal cells. PMID- 3039580 TI - On the neurotrophic action of melanocortins. PMID- 3039581 TI - Attention deficit disorder with hyperactivity (ADDH): the contribution of catecholaminergic activity. AB - An attention deficit disorder with hyperactivity in children (ADDH) is now recognized in most countries although diagnostic practices differ. Evidence is presented to show that the two cardinal symptoms of poor attentional performance and high motor activity may be functionally and causally separate. Both are temporarily relieved in a proportion of subjects that respond to psychostimulants. Beneficial treatment decreases nonadrenergic metabolism and normalizes variable levels of dopaminergic metabolism. Parallels are drawn with other clinical syndromes arising from changed catecholaminergic activity and with behavioral interpretations of the result of damage to the dorsal noradrenergic bundle and dopaminergic A10 nucleus. Prognosis of ADDH subjects after treatment remains poor. There may be a further defect of neurotransmitter metabolism in the ADDH syndrome. Research strategies are suggested based on the neurobiological correlates of the cognitive style of ADDH subjects and septal function in the animal model of the hypertensive rat. PMID- 3039582 TI - [Tumor necrosis factor--recent advances]. PMID- 3039583 TI - [Hydroxyproline levels in the serum and urine of workers exposed to benzene, hydrogen cyanide, sulfur dioxide and nitrogen dioxide]. PMID- 3039584 TI - Radical radiation therapy in inoperable non oat-cell carcinoma of the lung with particular emphasis on roentgenographic tumor response as prognostic factor. PMID- 3039585 TI - Ultrasonographic study of malignancies of extra-hepatic bile ducts. PMID- 3039586 TI - Exclusive use of radiation therapy in the treatment of non-small cell lung carcinoma. Evaluation of a series of 93 patients. PMID- 3039587 TI - [Clinical evaluation of gallium-67 scintigraphy in comparison with autopsy findings in the elderly, with particular reference to histological findings and classification of pulmonary cancer]. AB - A correlative study of autopsy findings and retrospective review of gallium scintigrams were performed in 106 elderly patients. Of the cases studied, 57% demonstrated positive gallium study in the present series. Histological correlation was undertaken in cases of lung cancer. Among them, squamous cell carcinoma showed the highest incidence of positive results (83%), whereas adenocarcinoma was the lowest (35%). There is no apparent correlation between subtypes of histological classification of adenocarcinoma and abnormal accumulation of gallium. However, abnormal accumulation of the nuclide seems to be rather related with interstitial reactions, namely fibrotic changes, lymphocyte infiltration and vascularization. PMID- 3039588 TI - Neuromuscular depression. PMID- 3039589 TI - The pyrolysis of cannabinoids. PMID- 3039590 TI - [Computed tomography of the soft tissues of the shoulder. 1. Synovial reactions]. AB - CT of the soft tissues of the shoulder without intra-articular contrast injections is useful for demonstrating effusions and synovial proliferation in the shoulder joint, in the bursa and in the synovial coverings of the long head of the biceps. The authors describe, for the first time, an intracapsular collection of fat in the posterior aspect of the capsule. This can be demonstrated via CT in 80% of the population. This fat line is displaced by even a few ml. of fluid in the presence of an effusion. PMID- 3039591 TI - [Sonographic measurement of the articular cartilage over the femoral condyles. A comparison with arthrography and computed tomographic pneumoarthrography]. AB - Sonographic evaluation of cartilage over femur condyles is equivalent to arthrography by CT. Circumscript lesions especially in the middle part of femur condyles can be seen easily by ultrasonography. PMID- 3039592 TI - [Extreme dilatation of the pulmonary trunk in severe long-term pulmonary hypertension]. PMID- 3039593 TI - [Hilar lymphoma as the 1st radiological manifestation of hypernephroma metastases. A case report]. PMID- 3039594 TI - Radiographic visualisation of seropositive rheumatoid arthritis in carriers of HLA-B27. AB - A group of 11 B27-positive, seropositive patients with rheumatoid arthritis was compared with 11 matched B27-negative seropositive patients. The radiographs of all limb joints, the sacroiliac joints, and the cervical spine were read blindly. Ten patients in each group were radiographed 2-6 times during observation periods of 3-13 years; one patient in each group was only examined once. The prevailing picture of both groups was that of progressive erosive rheumatoid arthritis, although two small differences were found: Erosions of the apophyseal joints of the cervical spine and slight periosteal new bone formation of the shoulder, hip, and knee regions occurred more often in the B27-positive than in the B27-negative group. PMID- 3039595 TI - [Neurofibromatosis and multiple nonossifying bone fibromas]. AB - Three patients with a combination of neurofibromatosis and multiple non-ossifying fibromas are presented. All patients possessed multiple osteolytic lesions with sclerotic margins, resembling non-ossifying fibromas. Two patients showed similar lesions in additional localizations. In two patients, non-ossifying fibroma was histologically verified. The simultaneous occurrence of neurofibromatosis and multiple non-ossifying fibromas can possibly be put down to a generalized mesodermal dysplasia. However, the definite explanation is unknown. PMID- 3039596 TI - [Results and complications of CT-guided puncture biopsies with a wide-lumen puncture needle]. AB - The results obtained in 187 CT-guided biopsies using a wide lumen needle are described. Biopsies in the thorax, abdomen, retroperitoneum and in the pelvis have been evaluated. A retrospective analysis of the material has shown an accuracy of 97.8% and sensitivity of 95.8%. The total complication rate was 8%. Amongst 120 biopsies in the thorax, 6.6% developed a pneumothorax. Therapeutic drainage was carried out in fifteen patients. PMID- 3039597 TI - [Mycoplasma pneumoniae encephalitis]. AB - Clinical, CT and, in one case, autopsy findings indicated a diagnosis of a severe necrotising encephalitis in two patients. Although usually herpes simplex virus is blamed for this form of encephalitis, it was possible to prove in these two patients that Mycoplasma was the causative agent of the disease. It is concluded that this organism can produce a serious disease in the central nervous system similar to that caused by herpes simplex. PMID- 3039598 TI - [Quantification of pulmonary alveolar microlithiasis in the CT]. AB - Pulmonary alveolar microlithiasis is a rare, familial disease with massive symmetrical intra-alveolar calcium deposition. Conventional CT findings and CT measurements with a dual energy technique were carried out in a 26-year-old patient suffering from this disease. The importance of the findings in the differential diagnosis and for estimating the progression and prognosis of the disease is discussed. PMID- 3039599 TI - [High-resolution computed tomography in malformations of the cochlear and vestibular organs]. AB - Twenty-one patients with congenital deformities of the petrous bones were examined by high resolution computed tomography. This allows an accurate description of the extent of the malformation in all parts of the cochlear and vestibular organs; only localised changes in the auditory ossicles, such as fixation of the stapes and changes in the soft tissue portions of the labyrinth, cannot be shown. Improved demonstration of soft tissues while using less radiation makes high resolution CT preferable to conventional polytomography. It is also easier to perform and provides axial projections which are essential for the elucidation of malformations of the middle and inner ear. PMID- 3039600 TI - [Radionuclide monitoring of gastrointestinal bleeding activity]. AB - Radionuclide monitoring was done in 50 patients to assess gastrointestinal bleeding, activity and location. Monitoring with 99m-Tc-in vivo-labelled erythrocytes was performed as sequential scintigraphy in increments of 1-2 hours up to 62 hours. 23 patients without active GI bleeding were correctly identified. 27 patients showed pathologic activities in abdominal blood-pool scintigraphy. In 25 patients peristaltic movement of these activities were seen--in each case we correctly diagnosed active GI bleeding. In 2 patients the activity stayed for a longer period in the same location--one patient had a liver hemangioma, the other patient had an aneurysm of the superior mesenteric artery. The great impact of radionuclide monitoring on diagnostic and therapeutic management of gastrointestinal bleeding is emphasized. PMID- 3039601 TI - [Rapid MR tomography of the liver. Technic and initial results]. AB - The fast-field-echo (FFE) sequence provides a new rapid imaging method for MR tomography. By using gradient echoes with shorter high frequency pulse repetitions than conventional sequences with 180 degrees echo pulses, it is possible to produce single cuts in 10 to 20 s with good signal-to-noise ratio. In this way the liver can be examined while respiratory movement artifacts are reduced. The possibility of obtaining T1- and T 2-weighted FFE images with good contrast resolution and the ability to determine relaxation times suggests a wide application of this rapid imaging procedure in the diagnosis of liver disease. PMID- 3039602 TI - [Fat/water separation in the NMR tomogram. The imaging of bone marrow reactions in degenerative intervertebral disk changes]. AB - We examined 410 patients with lumbar disc degeneration. In 23% fat suppressing IR sequences and phase contrast techniques displayed marrow reactions in subchondral bone adjacent to the affected discs. Only in some cases conventional MRI sequences were able to depict these marrow reactions. CT, plain X-ray and scintigraphy did not show marrow changes. Bacterial infection was excluded. Histological analysis showed substitution of haematopoietic marrow by fatty tissue, necrobiosis and increase in mucoid extracellular fluid. PMID- 3039603 TI - [X-ray morphology of osseous changes in pigmented villonodular synovitis of the joints]. AB - Pigmented villonodular synovitis (PVNS) of the joints is a rare, benign and generally monoarticular tumour-like soft tissue lesion of the synovium. The secondary bone changes, seen in plain films of 25 patients with PVNS are described. The main radiological features were paraarticular erosions, often with a thin sclerotic border. As a late manifestation these erosions tended to coalesce and to form large multicystic areas. These defects were observed particularly in the more tightly encapsulated joints, such as the hip, wrist, finger and toe. In 3 cases with involvement of the knee the lesions were monostotic. PVNS may be mistaken radiologically for other lesions such as malignant tumours or inflammatory disorders. The differential diagnosis is discussed. PMID- 3039604 TI - [MRT in neurofibromatosis and tuberous sclerosis]. AB - The relatively rare neurocutaneous syndromes may produce neurological symptoms during childhood. After closure of the fontanelle, the radiological methods of choice are CT and MRT. The value of these two methods is demonstrated by twelve examples (eight neurofibromatoses, four tuberous scleroses). CT is superior to MRT in being able to show calcification; it is therefore the method of choice for tuberous sclerosis. On the other hand, MRT may show foci of neurofibromatosis; these can hardly be seen on CT. In order to show cerebral lesions in these two conditions, T2-weighted sequences are necessary. PMID- 3039605 TI - [NMR tomographic studies in experimental renal vein ligation]. AB - Experimental ligation of the renal veins in rats indicate highly significant and characteristic changes within two hours, consisting of significant prolongation of T2 relaxation time in the cortex and T2 shortening in the medulla. In addition, there is a considerable increase in the size of the kidney, due to swelling of the cortex. T2 prolongation of the cortex is most marked between 30 hours and two to four days after ligation of the vein. In the following weeks there is a return to normal. T2 of the medulla at two to three weeks after ligation shows highly significant reduction compared with the normal side and, at this time, the size of the experimental kidney is significantly less than the opposite kidney. These results indicate that magnetic resonance tomography is a highly sensitive method for the early demonstration of renal vein thrombosis. PMID- 3039606 TI - [Prenatal sonography and computed tomography of fetal cerebral malformations]. AB - In three pregnant women, sonography and amniocentesis suggested cranial abnormalities of the foetuses. In view of the far-reaching consequences of such a diagnosis, CT was carried out to confirm the diagnosis. It was possible to show the intra-uterine abnormalities and the type of malformation in considerable detail. In one case an encephalocele was demonstrated, in the two others, an anencephalic foetus was shown. PMID- 3039607 TI - [Sonographic imaging and assessment of stomach wall changes]. AB - By the use of a special method of investigation, the true value of percutaneous echography was investigated using 28 patients with primary gastric neoplasms and 3 patients with secondary infiltration of the gastric wall, for the proof and the judgement of the tumors, their extension and their metastases. Tumorous gastric wall thickening could be demonstrated in 87% (27/31), tumor invasion in neighbouring structures in 85% (12/14) and lymph nodes metastases in 66% (10/15). In all patients, who had liver metastases (4) or ascites (5) they were demonstrable. The sonographic appearance of the different gastric wall changes are described and discussed. PMID- 3039608 TI - Ultrasound in acute and chronic cholecystitis. AB - Sonograms of 412 consecutive patients with operatively and histologically proven acute or chronic cholecystitis were reviewed retrospectively. 267 of them had been operated on in an acute phase of gallbladder disease and 145 had undergone an elective operation. 236 patients had an operative and/or histological diagnosis of acute cholecystitis and 176 both operatively and histologically confirmed chronic cholecystitis. A thickened gallbladder wall was seen in 80% of acute cases, wall sonolucency in 39%, dilated gallbladder in 60%, sludge in 26% and stones in 75%, the corresponding proportions of the chronic cases being 18%, 4%, 12%, 13% and 93%. A contracted gallbladder with stones was seen in 15% of the chronic and 1% of the acute cholecystitis cases. 90% of the acute cases had two or more sonographic abnormalities, whereas 70% of the chronic cases had only one abnormality, most frequently gallstones. Only one normal sonographic finding was recorded in each group. PMID- 3039609 TI - [Microangiographic findings in experimental intestinal anastomoses]. AB - Microangiography of anastomosis at the rat colon and the pig jejunum did reveal the postoperative state of vessel regeneration. Findings for evaluation were avascular areas, dislocation and dilatation of vessels, vessel break, vessel anastomosis to the opposite site and to adhesions as well as the complete regeneration of the vessel architecture. A suture technique leading to correct apposition of the wound edges without destroying the circulation will be followed by an almost complete regeneration of the vessel architecture. PMID- 3039611 TI - [Ganglioneuroma of the lesser pelvis]. PMID- 3039610 TI - [Neuroblastoma IV-S. A case contribution on hepatomegaly]. PMID- 3039612 TI - [The colon as a perirenal space-occupying structure in ultrasonic examination]. PMID- 3039613 TI - [Lyme disease: the computed tomographic and MR tomographic images]. PMID- 3039614 TI - [Computed tomographic diagnosis of an aneurysm of an arteria lusoria]. PMID- 3039615 TI - [Antidepressants and anesthesia]. PMID- 3039617 TI - [Pulmonary emphysema, liver cirrhosis and primary hepatocarcinoma in alpha 1 antitrypsin deficiency]. PMID- 3039616 TI - [Veno-venous bypass in a special case of liver transplant]. PMID- 3039618 TI - [Current status of after-care cures in oncologic patients]. AB - Some 40 accredited aftercare clinics are available for inpatient cancer aftercare, since 1981 also for immediately post-acute treatment courses. These clinics are used for both, the frequent physical treatment aftermath from oncological diseases in the otological or gynaecological field, and for psychic stabilization when psychosocial problems arise. Participation in autogeneous relaxation training, yoga, and/or psychotherapeutic services are found to entail improved disease coping. Concrete patient care expectancies primarily center around movement therapy, lymphatic drainage (in breast carcinoma), speech therapy (in laryngectomees), pain control, but are also directed at psychosocial care in terms of achieving" inner calmness and relaxation". Given the age distribution seen, increasing the quality of life takes priority over restoration of the working capacity. PMID- 3039619 TI - [Effect of the addition of a fiber-rich concentrate to a hay-based diet, offered at 2 dietary levels to adult rabbits. 1. Mean retention time of digesta]. AB - The influence of adding a fibre rich concentrate feed (wheat bran or beet pulp) to a hay-based diet (orchard grass-lucerne mixture; 50/50) upon the rate of passage of digesta of these two fractions (hay and concentrate) was measured in three groups of 8 adult doe rabbits (White New Zealand x California). The three diets composed of either 90% hay, or hay + bran or hay + pulp were offered at two levels of feeding, i.e. ad libitum and 2/3 ad libitum. The retention time of the two main fractions was measured by means of two tracers, i.e. ytterbium-169 bound to hay and cerium-141 to bran or pulp. Animals fed on the 90% hay diet excreted more than 90% of the tracer during the first 24 h. The mean retention time (MRT) of this diet was the shortest (on an average 14.8 h). The addition of pulp or bran increased the MRT of the diet by 2 and 4 h, respectively. Feed restriction led to a 5 to 7 h earlier period of caecotrophy with a subsequent lengthening of the MRT by 2 to 4 h according to groups. The two hay + "concentrate" diets gave almost the same 169Yb and 141Ce excretion curves. Thus, the mean retention time seems to be the same for digesta from hay and "concentrate" (non significant deviations). PMID- 3039620 TI - Antibody and complement mediated lysis of felid herpesvirus 1 infected cells in vitro. AB - The lysis of cells infected by felid herpesvirus 1 (FHV) by feline anti-FHV antibody and complement was demonstrated. Lytic activity was sensitive to dilution of both antibody and, especially, complement. It was first detected within 10 to 20 minutes, increased rapidly during the next 30 minutes of incubation and then rose more slowly in a linear manner. Using standard antibody and complement concentrations and assay duration, it was shown that FHV infected cells underwent significant (P less than 0.05) lysis from eight hours after infection in a system in which FHV-specific membrane antigen was first detected at three hours after infection and spread of FHV by the intracellular route began eight to nine hours after infection. The ability of antibody and complement to reduce FHV spread in this system was demonstrated by a significant (P less than 0.05) reduction in FHV plaque numbers, although the restriction of spread was not absolute. Chelation of divalent cations and heat inactivation of complement factor B revealed that the lytic system was dependent on factor B and Mg2+ but not Ca2+, suggesting involvement of the alternative pathway of complement activation. PMID- 3039621 TI - Effect of acyclovir on the replication of turkey herpesvirus and Marek's disease virus. AB - The toxicity of acyclovir for chick embryo fibroblasts and its effect on the replication of turkey herpesvirus (strain FC 126) and Marek's disease virus (strain HPRS 16) multiplied on fibroblast culture was studied. The influence of using acyclovir on the development of the tumour process in birds infected with a virulent Marek's disease virus was also determined. Acyclovir used in doses below 12.5 micrograms ml-1 proved to be nontoxic for chick embryo fibroblast culture. It inhibited in vitro replication of turkey herpesvirus and Marek's disease virus. It was also shown to diminish the development of tumours in birds infected with Marek's disease virus. PMID- 3039622 TI - Effects of megestrol acetate on glucose tolerance and growth hormone secretion in the cat. AB - Long-term administration of relatively high therapeutic dosages of megestrol acetate to cats produced a progressive deterioration in glucose tolerance, with a significant (P less than 0.05) increase in mean fasting plasma glucose concentrations and decrease in mean plasma glucose clearance rates after six and 12 months of treatment. There appeared to be no relationship, however, between the development of glucose intolerance and circulating growth hormone (GH) concentrations in the cats of this study, since no significant rise in plasma GH concentrations was detected during the 12 month period of megestrol acetate treatment. Administration of megestrol acetate also produced a progressive decrease in both resting plasma cortisol concentrations and cortisol concentrations after ACTH stimulation. Three months after discontinuation of megestrol acetate, the elevated fasting plasma glucose concentrations, decreased glucose clearance rates and subnormal plasma cortisol concentrations all returned to normal pretreatment values, indicating resolution of glucose intolerance and hypoadrenocorticism. The results of this study demonstrate that administration of megestrol acetate to cats can produce a state of moderate to severe glucose intolerance, which is usually reversible after cessation of treatment. Although the exact mechanism of the glucose intolerance and overt diabetes mellitus induced by progestagen treatment of cats remains unclear, it is likely that these alterations in glucose metabolism result primarily from the glucocorticoid activity intrinsic to megestrol acetate. PMID- 3039623 TI - Demonstration in live chickens of the carrier state in infectious laryngotracheitis. AB - Two separate groups of nine-week-old specific pathogen free cockerels maintained in isolation were infected with a field strain of infectious laryngotracheitis (ILT) virus, either by intratracheal or combined intranasal and supraconjunctival inoculation. Birds were monitored for virus shedding from five sites on alternate days during the acute phase and three times weekly until week 17. They were then treated with cyclophosphamide on three consecutive days and thereafter swabbed daily. During the acute phase clinical signs were observed and virus was recovered from ocular and nasal sites for up to six to eight days. Initially after the acute phase no virus could be detected. However, from the seventh week after infection intermittent, apparently spontaneous shedding was detected in four of five birds in each group. There was no clear effect of cyclophosphamide treatment on re-excretion patterns, possibly because of the high levels of virus shedding already occurring. Thus, a carrier state for ILT virus has been demonstrated experimentally in live clinically recovered birds. PMID- 3039624 TI - The significance of plant transposable elements in biological processes. PMID- 3039625 TI - [Phlebosclerosis and arteriosclerosis with different pathogenesis]. PMID- 3039626 TI - Rate of secretion of lung surfactant before and after birth. AB - Using electron microscopic techniques, we defined the percentage of intracellular lamellar bodies (LB) in the process of extrusion from type II cells, designated that as an index of secretion of surfactant, and studied the rate of release of surfactant in the lungs of fetal and newborn rats. The index of secretion increased twice in perinatal period. The most dramatic increase occurred during labor which consequently declined to fetal level within 15 min of birth. The second rise began at 30 min after birth and reached maximum in 2 h. Thereafter, it gradually declined reaching its lowest at 24 h. The number of LB/type II cell decreased with an increase in the rate of release and began to rise at about 1-2 h after increase in the rate of release. It is speculated that the rise in secretion rate during labor is in response to the change in circulating hormone levels, and that for after birth in response to lung distension and air ventilation. We also used this method of analysis in lavaged fetal lungs to test our unconfirmed belief that lung lavage promotes release of intracellular LB. The results, consisting of a 5-fold increase in the index of secretion and a 20% reduction in the number of LB/cell in lavaged lungs, were in favour of the notion. This should be taken into consideration when lung lavage is used to obtain 'extracellular surfactant'. PMID- 3039627 TI - [Serum variations of the antibody titer for some viruses in sudden cochleovestibular disease]. PMID- 3039628 TI - Carcinomas of the lung with neuroendocrine differentiation. AB - While there are examples of each of the histologic types of bronchogenic carcinoma in which ectopic hormone production has been demonstrated, three groups of lung cancers manifest this type of differentiation with great regularity. These are the carcinoid, atypical carcinoid, and the small cell carcinoma, which have been called neuroendocrine carcinomas of the lung. The carcinoids are neoplasms with a heterogeneous array of histologic appearances that share relatively uniform nuclear features, the absence of both tumor necrosis and mitotic figures, and a good prognosis. Neuroendocrine differentiation is demonstrable with such regularity in carcinoids that the diagnosis is not considered tenable in the absence of at least one of the following features: argyrophilia, the demonstration of neuron-specific-enolase or neuropeptides by immunohistochemistry or other techniques, or the demonstration of numerous dense core membrane-bound granules, usually 150 to 250 nm in diameter, by electron microscopy. The atypical carcinoids (well or moderately differentiated neuroendocrine carcinomas) share the neuroendocrine differentiation of the carcinoids and many of their histologic features, but are distinguished by the presence of tumor necrosis, more anaplastic large cell nuclei with numerous mitotic figures, and a distinctly worse prognosis. They are a heterogeneous lot with some similarities to carcinoids, small cell carcinomas, and large cell or adenocarcinomas. The demonstration of at least one of the neuroendocrine features listed above is considered necessary for this diagnosis. Small cell carcinoma is also a heterogeneous group of neoplasma primarily distinguished by their finely granular chromatin pattern which correlates with a much worse prognosis but a higher likelihood of response to chemotherapy and radiotherapy. Many small cell carcinomas share the neuroendocrine differentiation of the carcinoids or atypical carcinoids, but some do not and the demonstration of these features is not a requisite for inclusion in the small group. The morphologic demonstration of neuroendocrine differentiation may be more difficult in small cell carcinomas, but they more frequently produce clinically important amounts of ectopic neuropeptides. While the term neuroendocrine carcinoma of the lung includes several quite different entities and does not by itself convey histogenetic or prognostic implications, the demonstration of neuroendocrine differentiation in pulmonary neoplasms is an important procedure when combined with the recognition of other cytologic and histologic features. PMID- 3039629 TI - Large cell carcinoma of the lung. AB - The diagnosis of large cell carcinoma of the lung on the basis of hematoxylin and eosin (H & E) stains represents a heterogeneous group, both endodermally and non endodermally derived, totalling about 16% of lung cancers. By the use of special stains, such as Kreyberg and Grimelius stains, this percentage can be sharply reduced, leaving a core group of about 10%, including giant cell and clear cell variants. This core group may show a variety of ultramicroscopic organelles that are insufficiently developed to alter the light microscopic characteristics on which the diagnosis depends. In a study of 1000 slides, large cell carcinoma constituted 6% of the squamous cell carcinoma slides and 21% of the adenocarcinoma slides, which was reduced to 3% by mucin stains. Giant cells predominated in 5% and clear cells in 10% of large cell carcinoma slides. Large cells also occurred in 12% of small cell carcinoma slides. PMID- 3039630 TI - The intravascular bronchioloalveolar tumor and the sclerosing hemangioma of the lung: misnomers of pulmonary neoplasia. AB - Two very different uncommon pulmonary tumors continue to bear the misleading names originally given them. The intravascular bronchioloalveolar tumor (IVBAT) is not an epithelial tumor, but is an incurable multifocal primary pulmonary angiosarcoma with a variable and often indolent clinical course. Conversely the sclerosing hemangioma of the lung is not a vascular tumor, but is a solitary benign epithelial tumor of uncertain histogenesis with a complex architecture. PMID- 3039631 TI - Human corticotroph cell adenomas. AB - Sixty-one pituitary corticotroph adenomas from 47 patients with Cushing's disease, 10 with Nelson's syndrome, and four eucorticoid patients were studied by light microscopy, immunoperoxidase, and electron microscopy. Seventy nine percent of all tumors and 70% of Nelson's cases were microadenomas, sometimes minute. A contiguity between the posterior lobe and the adenoma was seen in ten cases. Spontaneous infarction of the tumor with remission of Cushing's syndrome occurred in one case. Light microscopy revealed that the adenoma cells were basophilic and contained PAS-positive granules also staining with Herlant tetrachrome and lead hematoxylin. The granules stained positively with antiserum to adrenocorticotrophic hormone (ACTH), beta-lipotropic hormones (beta-LPH) and beta endorphin. The most characteristic ultrastructural finding was the presence of perinuclear bundles of microfilaments found in all our cases. Oncocytic changes were seen in three tumors. Four silent corticotroph adenomas, two of them originally microadenomas that had enlarged to enclosed adenomas while being treated with bromocriptine for hyperprolactinemia and one a large diffuse invasive tumor, did not differ in their microscopic, immunocytological, or ultrastructural features. PMID- 3039632 TI - Plurihormonal pituitary adenomas. AB - Plurihormonal adenomas of the pituitary, ie, tumors that engage in the production of unusual combinations of hormones, represent approximately 10% to 15% of all adenomas. Such tumors comprise in excess of 50% of adenomas in the setting of acromegaly and occur with somewhat greater frequency in childhood and adolescence than in adulthood. Eight percent are associated with multiple endocrine neoplasia, type I. The most common variant of plurihormonal adenoma produces growth hormone, prolactin, and one or more glycoprotein hormones, the most common being TSH. Clinical effects most often reflect the presence of growth hormone, and to a lesser extent, prolactin cells; expression of glycoprotein hormone production is rare. The tumors are more often macroadenomas (80%) than microadenomas (20%) and demonstrate gross invasion in 50% of cases. Plurihormonal adenomas may be ultrastructurally monomorphous, bimorphous, or trimorphous; thus, one morphologic cell type may elaborate several hormones. PMID- 3039633 TI - Ultrasonically guided percutaneous aspiration biopsy of the pancreas. AB - One hundred and twenty-six patients had fine needle aspiration of the pancreas under ultrasonic guidance. The aspirates of 66 patients contained malignant cells. Two of 38 patients who had negative cytology subsequently were diagnosed at laparotomy as having carcinoma of the pancreas. The results of this study show that the sensitivity of the cytologic method was 97%, and the specificity and predictive value were 100%. PMID- 3039634 TI - Transthoracic aspiration cytology of pulmonary lesions. AB - In a seven-year period (1974 to 1981), 1,390 patients had transthoracic aspiration biopsy for radiologically suspected pulmonary neoplasms. Of the 1,209 proven malignant neoplasms, 1,059 (88%) were cytologically diagnosed as malignant. There were only two (0.2%) false-positive diagnoses both due to florid bronchoalveolar hyperplasia associated with granulomatous disease. Cytologic and histologic correlations for primary lung cancers were 92% for adenocarcinoma, 87% for oat cell (small cell) carcinoma, and 83% for epidermoid carcinoma. Characteristic cytomorphology of pulmonary carcinomas, some metastatic neoplasms, and inflammatory lesions are described. PMID- 3039635 TI - Aspiration cytology of salivary glands. AB - Although mass lesions of the salivary glands are readily accessible to examination by fine-needle aspiration, the use of this modality has been limited. In part, this may be related to the difficulty differentiating between benign and malignant neoplasms in some cytologic specimens. Marked atypia in reactive non neoplastic epithelium also could result in a false-positive diagnosis. In addition, aspiration of hypocellular material from cystic neoplasms, eg, well differentiated mucoepidermoid carcinoma, may lead to false-negative cytologic reports. However, the diagnostic specificities claimed by a number of authors for this method are excellent. Furthermore, aspirates of certain specific neoplasms may yield highly distinctive cellular samples, such as the uniform tumor cells and extracellular hyaline spheres in many adenoid cystic carcinomas. Another example is the characteristic transition between the epithelial and myoepithelial cells of pleomorphic adenomas, which may contain prominent myxoid matrical material. With the increasing recognition of such features, the reported levels of diagnostic accuracy are improving. PMID- 3039636 TI - Small cell carcinoma of the lung: an overview. PMID- 3039637 TI - Non-small cell lung cancer: issues in diagnosis, staging and treatment. PMID- 3039638 TI - [Viral hepatitis in a rural area in Chile]. PMID- 3039639 TI - [Treatment of non-small cell cancer of the lung with a combination of cisplatin VP-16 and mitomycin C]. PMID- 3039640 TI - [Primary carcinoma of the liver]. PMID- 3039641 TI - [A case of pancreatic cancer with invasion to the wall of the intrahepatic bile duct]. AB - Presented here is the case of a 69-year-old woman with pancreatic cancer. Efforts were made to discover the origin of the cancer but without success. Tumor nodules were scarcely observed in the pancreas macroscopically at autopsy, while histological examination of the pancreas revealed the presence of a well differentiated adenocarcinoma (duct cell carcinoma). The invasion to the wall of the intrahepatic bile duct was also observed microscopically. PMID- 3039642 TI - [Leg pains of vascular origin]. PMID- 3039643 TI - [Bronchiolo-alveolar carcinoma complicating systemic scleroderma under long-term treatment with factor XIII]. AB - A 52-year old woman with systemic scleroderma had been treated since 1978 with factor XIII. The skin lesions had improved, and respiratory lesions had become stabilized. In 1984, she developed a bronchiolo-alveolar carcinoma which was treated by surgery. The authors emphasize that factor XIII is well tolerated and has beneficial effects on the skin and probably also on the lungs, without tachyphylaxis; however, the possibility of a bronchioloalveolar carcinoma occurring cannot be ruled out. PMID- 3039644 TI - The role of neurokinin A and calcitonin gene-related peptide in the mucociliary defence of the rabbit maxillary sinus. AB - Substance P (SP) released from sensory C-fibres in the airways increases the mucociliary (m.c.) activity in the rabbit maxillary sinus. The purpose of the present study was to investigate the m.c. effects of two other neuropeptides, coexisting with SP in sensory neurones, neurokinin A (NKA) and calcitonin gene related peptide (CGRP). NKA increased the m.c. activity dose-dependently (dose range 0.1-5.0 micrograms/kg) the maximum increase being 33.6 +/- 6.0%. The effect was inhibited by pretreatment with the tachykinin antagonist (D-Pro2, D Trp7,9)SP, but not with atropine or hexamethonium. Thus NKA released from sensory C-fibres may contribute to the non-cholinergic increase of m.c. activity observed after C-fibre stimulation. In contrast CGRP did not influence the m.c. activity. Neither did it influence the responses to NKA or SP. It is concluded that CGRP is unlikely to be involved in the control of m.c. function. PMID- 3039645 TI - Immunosuppression in progressive multiple sclerosis with high dose intravenous cyclophosphamide and monoclonal antibodies. AB - Our work with multiple sclerosis has demonstrated a favorable effect on the course of chronic progressive multiple sclerosis in two-thirds of patients treated with cyclophosphamide/ACTH. However, alternative methods of therapy, or repeated treatments with cyclophosphamide and ACTH are required for longer term control of the illness. We are attempting to assess the efficacy of outpatient maintenance cyclophosphamide, but do not as yet have any data to support the use of this form of treatment. Cyclophosphamide and ACTH can be given to multiple sclerosis patients without significant serious toxicity. Monoclonal antibody therapy of multiple sclerosis is in its beginning stages. The treatment appears to be safe and has produced some interesting data indicating that rapid entry of some labeled lymphocytes into the nervous system can be measured by this technique. Since these are phase one pilot trials designed to assess immune parameters and only small numbers of patients have been treated, no clinical results have been obtained. No adverse effects of monoclonal antibody therapy were observed. PMID- 3039646 TI - Detection of free radical intermediates in the oxidative metabolism of carcinogenic hydrazine derivatives. AB - Hydrazine derivatives are widely used in agriculture, in industry, as rocket propellants, and in medicine. Hydrazines also occur naturally in tobacco and mushrooms. Many hydrazines tested in animal studies appear to be carcinogenic and induce tumors in various target tissues in mice, hamsters, and rats. The use of hydrazine derivatives in humans is often complicated by adverse side-effects such as liver injury and rheumatoid arthritis. A number of studies have demonstrated that hydrazine derivatives are activated to reactive intermediates, such as free radicals, through a variety of cellular oxidative metabolic pathways. The aim of this work is to demonstrate the occurrence of free radical intermediates during the metabolic activation of various hydrazine derivatives and to characterize the enzymatic system(s) responsible for the activation to free radical species. The hydrazines studied are acetylhydrazine, isoniazid, isopropylhydrazine, iproniazid, methylhydrazine, 1,1-dimethylhydrazine, and 1,2-dimethylhydrazine. The model systems chosen are those of rat liver microsomes and isolated hepatocytes. Free radical intermediates have been demonstrated by the electron spin resonance spectroscopy coupled to spin trapping technique. The activation mechanism has been characterized using inhibitors of the mixed function oxidase system and of the FAD-dependent oxygenase system. Glutathione was able to scavenge, with high efficiency, the free radicals produced. PMID- 3039647 TI - [Expression of T-lymphocyte antigens by serum B lymphocytes in rheumatoid polyarthritis]. AB - Molecules recognised by the monoclonal antibodies Leu 1 - OKT 1 - Mid 5 (CD5 molecules) were originally shown on human T cells and are equivalent to Lyt-1 molecules in the mouse. These surface molecules are detectable on some human B cells, but are weakly expressed. We have therefore tested the effects of the tumour-promoter, phorbol myristic acetate (PMA) on expression of these molecules on B cells. Sheep red blood cell rosette-depleted lymphoid organ or blood cells from 8 controls and 10 rheumatoid arthritis (RA) patients were examined fresh and cultured with PMA for 48 hours and the cells stained with Leu 1 or Mid 5 by indirect immunofluorescence. Cells were analysed by UV microscopy or flow cytometry. More RA blood B cells were positive for Leu 1 than normals before culture. Following incubation with PMA, the percentage of positive cells increased in all cell populations (1-49% in RA patients and 5-26% in controls). There was no difference in the expression of Leu 1 on Kappa--or lambda--positive cells before and after culture in PMA. Leu 1 positive cells were under represented in Epstein-Barr virus-driven cell lines. PMID- 3039648 TI - [Nature and immunogenicity of the cytoskeleton]. PMID- 3039649 TI - [Chin neuropathy in myeloma]. PMID- 3039650 TI - Reactive oxygen species selectively deplete normal T lymphocytes via a hydroxyl radical dependent mechanism. AB - Chronic inflammatory synovitis is characterized by both lymphocytic infiltrates and persistent polymorph exudates. Activated polymorphs release reactive oxygen species (ROS) during inflammation, but the contribution that these make to the lymphocyte abnormalities associated with RA has been little studied. We therefore investigated the cytotoxic effects of the reactive oxygen species on human peripheral blood mononuclear cells (PBMC). PBMC were exposed to RPMI 1640 medium previously irradiated for up to 60 min. Consistent dose-dependent killing was observed at 24 h. Antioxidant studies indicated that H2O2 was the effective species. Catalase, which specifically degrades H2O2, gave almost total protection against cell death, while superoxide dismutase (SOD), thiourea, and mannitol were largely ineffective. Addition of exogenous H2O2 caused an identical pattern of cell death to that observed with irradiated medium. PBMC cultures supplemented with desferrioxamine (a ferric iron chelator) also gave significant protection, suggesting that H2O2 mediated its effects via OH radicals. Analysis of lymphocyte subpopulations showed that ROS caused a selective depletion, depending on the level of H2O2 present. Low levels induced a specific loss of CD8+ cells, while higher concentrations caused significant loss of CD4+ T cells as well. sIg+ B cells were unaffected at either concentration. This selective lymphotoxic effect of ROS may be of considerable importance in the pathogenesis of autoimmune inflammatory disease. PMID- 3039651 TI - A prospective study on infection with cytomegalovirus in renal allograft recipients immunosuppressed with cyclosporine A and low dose prednisone. AB - To investigate the course of infection with cytomegalovirus (CMV) in renal allograft recipients treated with cyclosporine A, 10 patients were followed for 1 year after transplantation. Virus cultures from blood, urine and throat washings were performed employing a quantitative technique. Complement-fixing and IgM antibodies to CMV were measured at scheduled intervals. The incidence 90% and course of CMV infections were found not to differ from those reported in patients receiving conventional immunosuppressive therapy. The quantitative virus cultures showed a consistent pattern with viremia most prominent at the beginning of an infection, and the highest concentration found in the one patient who developed symptoms of viral disease. It is suggested that information about the concentration of virus in a specimen will improve the diagnostic value of virus cultures in this group of patients. PMID- 3039652 TI - Comparison of four serological tests for the detection of specific immunoglobulin M in cord sera of infants congenitally infected with cytomegalovirus. AB - Four serological tests (3 immunoassays using enzyme-labelled antigen and 1 radioimmunoassay) were compared as regards the detection of cytomegalovirus (CMV) immunoglobulin M in cord sera. 68 cord sera from infants congenitally infected by CMV were included in the study. The infections were primarily diagnosed by virus isolation close to birth. Four laboratories in 3 countries were involved, each laboratory using its own or a commercial test. Of the sera tested in the different laboratories 50-80% were found to be reactive. Both qualitatively and quantitatively there was a good correlation between the 3 enzyme-immunoassays. The RIA results differed to some extent from the enzyme tests as regards the quantification of IgM. The advantage of prospective IgM screening in undiluted cord sera followed by confirmatory virus isolation test in the neonatal period in IgM-positive cases is discussed. PMID- 3039653 TI - Fulminant course of infectious mononucleosis with virus-associated hemophagocytic syndrome. AB - A fatal case of infectious mononucleosis due to serologically verified Epstein Barr virus infection in a previously healthy 30-year-old man is presented. The clinical course was characterized by severe prostration, persistently high spiking fever, and continuous development of enlarged lymph nodes. Hematologic examination revealed peripheral leukopenia and thrombocytopenia, and in the bone marrow an increased number of benign histiocytes showed marked hemophagocytosis. At autopsy abnormal lymphoid infiltrates were present in several tissues. The pathogenesis of this infection-associated hemophagocytic syndrome is discussed in terms of the possibility of an impaired immune response to infectious agents. PMID- 3039654 TI - Acute renal failure caused by polymyxin B containing ointment. AB - A case is presented in which a 67-year-old man suffering from non-insulin dependent diabetes mellitus, after being treated with polymyxin B containing ointment for leg ulcers, developed acute renal failure. After a period of 13 days during which the patient was treated with peritoneal dialysis 5 times, renal function returned. At discharge creatinine-clearance was 12 ml/min. Treating large ulcers with polymyxin B may be dangerous. PMID- 3039655 TI - Surgical management of tracheal tumours. AB - The annual incidence of primary tracheal tumours in Sweden is less than 1 per million population. Five cases of malignant tracheal neoplasm treated with segmental resection and primary reconstruction are described. Exploration and mobilization of the trachea were performed via right thoracotomy. Suprahyoid laryngeal release was also done in two cases, using a cervicomediastinal approach. The length of resected segment in these cases was 6 and 7 cm. High frequency positive-pressure ventilation was used in four of the five cases and greatly facilitated the operation. Recovery was uneventful. Adenoid cystic carcinoma was too extensive for extirpation in one case, but 4 months after radiotherapy a 7 cm tracheal segment with residual tumour was removed; 3 years later the patient is well. There was no stenosis or other late complication and no local recurrence in the long-term survivors. No vocal paralysis occurred. The two patients with laryngeal release had remarkably little and transitory dysphagia. Technical problems are discussed and conclusions are presented. PMID- 3039656 TI - The choice of opioid receptor subtype in isolated preparations by ohmefentanyl. AB - Ohmefentanyl has significant inhibitory actions on the electrically evoked contractions of guinea pig ileum and mouse vas deferens. Their IC50 values are 0.15 nM and 0.89 nM, respectively. In the guinea pig ileum and the mouse vas deferens, both mu-antagonist naloxone and (mu + kappa)-antagonist Mr. 2266 antagonize readily the agonist action of ohmefentanyl, indicating that ohmefentanyl acts on mu-receptors in the guinea pig ileum and the mouse vas deferens. Like other mu-agonists, ohmefentanyl has agonist activity but no antagonist action in the rat vas deferens. In the rabbit vas deferens, the activity of ohmefentanyl is very weak. The IC50 value is 149 nM. These results further indicate that ohmefentanyl is a potent and selective mu-agonist. PMID- 3039657 TI - Cell-autonomous determination of cell-type choice in Dictyostelium development by cell-cycle phase. AB - The developmental fate of individual cells has been examined in a system that allows Dictyostelium discoideum cells to differentiate in the absence of aggregation. The results show that the propensity of single amoebae to differentiate into either prespore or prestalk cells occurs by a cell-autonomous mechanism dependent on the cell's position in the cell cycle at the initiation of development. Cells that divide between approximately 1 1/2 hours before and approximately 40 minutes after the differentiation-inducing starvation become prestalk, whereas cells dividing at other times become prespore cells. These results suggest mechanisms by which an initial proportioning of the two cell types within the aggregate is achieved. PMID- 3039658 TI - Stable integration and expression of a bacterial gene in the mosquito Anopheles gambiae. AB - Foreign DNA was successfully introduced into the germline of the African mosquito vector of malaria Anopheles gambiae. Stable integration of genes into the germlines of insects had been achieved previously only in Drosophila melanogaster and related species and required the use of the P element transposon. In these experiments with Anopheles gambiae, the plasmid pUChsneo was used, which contains the selectable marker neo gene flanked by P element inverted repeats. Mosquitoes injected with this plasmid were screened for resistance to the neomycin analog G 418. A single event of plasmid insertion was recovered. Integration appears to be stable and, thus far, resistance to G-418 has been expressed for eight generations. The transformation event appears to be independent of P. PMID- 3039659 TI - The fos gene as "master switch". PMID- 3039660 TI - Ouabain resistance conferred by expression of the cDNA for a murine Na+, K+ ATPase alpha subunit. AB - The molecular basis for the marked difference between primate and rodent cells in sensitivity to the cardiac glycoside ouabain has been established by genetic techniques. A complementary DNA encoding the entire alpha 1 subunit of the mouse Na+- and K+-dependent adenosine triphosphatase (ATPase) was inserted into the expression vector pSV2. This engineered DNA molecule confers resistance against 10(-4) M ouabain to monkey CV-1 cells. Deletion of sequences encoding the carboxyl terminus of the alpha 1 subunit abolish the activity of the complementary DNA. The ability to assay the biological activity of this ATPase in a transfection protocol permits the application of molecular genetic techniques to the analysis of structure-function relationships for the enzyme that establishes the internal Na+/K+ environment of most animal cells. The full-length alpha 1 subunit complementary DNA will also be useful as a dominant selectable marker for somatic cell genetic studies utilizing ouabain-sensitive cells. PMID- 3039661 TI - The maize transposable element Ds is spliced from RNA. AB - In some instances, insertion of maize transposable elements into exons does not result in the total loss of enzymatic activity. In other instances, messenger RNAs of wild-type size are encoded by genes known to contain the maize transposable element Dissociation (Ds) in exons. To understand how Ds is processed from RNA, a study was made of transcripts encoded by two alleles of the maize waxy (wx) gene containing Ds insertions in exon sequences. The analysis was carried out in strains where the Ds element could not excise from the wx gene. Despite insertions of 4.3- and 1.5-Ds elements, the predominant transcripts encoded by these two genes were wild type in size. For both alleles, DNA sequencing of complementary DNAs revealed that the Ds elements had been spliced in a similar manner. Splicing was accomplished by the utilization of multiple 5' donor splice sites in the Ds termini and a 3' acceptor site within the wx gene adjacent to the Ds element. The net effect in both cases was the removal of most of the Ds element from the messenger RNA. PMID- 3039662 TI - Functional interaction and partial homology between human immunodeficiency virus and neuroleukin. AB - Dementia is common in patients with AIDS, but the mechanism by which the human immunodeficiency virus type 1 (HIV-1) causes the neurological impairment is unknown. In this study the possibility that an antigen of HIV-1 suppresses neuronal responses to neurotrophic factors was examined. Both HIV-1 and a related retrovirus, simian immunodeficiency virus (SIV), inhibited the growth of sensory neurons from chick dorsal root ganglia in medium containing neuroleukin (NLK) but not in medium containing nerve growth factor. An unrelated type D retrovirus, simian acquired immunodeficiency syndrome virus, did not affect the growth of neurons in the presence of either neurotrophic factor. The inhibition by HIV-1 of neuron growth in the presence of NLK was found to be due to the gp120 envelope glycoprotein. Regions of sequence homology between gp120 and NLK may account for this inhibitory property of gp120 and functional interactions between gp120 and NLK may be important in the pathogenesis of the AIDS dementia complex. PMID- 3039663 TI - Solo actions of AIDS virus coat. PMID- 3039664 TI - Functional imaging of the brain. AB - The radionuclide tracer method is unique amongst all other imaging methodologies in its ability to trace organ or tissue function and metabolism. It derives this advantage from the nature of the signal used for image generation, and its single interaction with the organ or system under examination. Physical processes such as electron or proton density assessment or resonance, edge identification, electrical or ultrasonic impedence, do not pertain to the image generation process in nuclear medicine, and if so, only in a rather secondary manner. The nuclear medicine imaging study is primarily a study of the chemical nature, distribution and interaction of the tracer/radiopharmaceutical utilised with the cellular system which requires investigation: the thyroid cells with sodium iodide, the recticular endothelial cells with colloidal particles, the adrenal medulla cells with metaiodobenzylguanidine, and so on. In the two most recent areas of nuclear medicine expansion, oncology (with labelled monoclonal antibodies) and neurology and psychiatry (with a whole new series of lipid soluble radiopharmaceuticals), specific cell systems can also be targeted and hence imaged and investigated. The study of structure as masterly performed by Virchow and all his successors over more than a century, is now definitely the prerogative of such imaging systems which excel with spatial and contrast resolution (x-ray computed transmission tomography, nuclear magnetic resonance imaging, diagnostic ultrasound). However the investigation of function and metabolism (as performed by Claude Bernard, Georg von Hevesy, and so many others), has clearly passed from the laboratory animal protocol and experiment to the direct investigation in man, this being the achievement of the radionuclide tracer methodology. In this article, we review present interest and developments in that part of nuclear medicine activity which is aimed at the study of the neurological or psychiatric patient. PMID- 3039665 TI - Thallium 201/99mTc parathyroid subtraction scintigraphy of the neck: single area of increased thallium uptake. PMID- 3039666 TI - Thallium 201/99mTc parathyroid subtraction scintigraphy of the mediastinum. PMID- 3039667 TI - Reovirus type 3 and neonatal cholestasis. PMID- 3039668 TI - Systemic disorders associated with neonatal cholestasis. PMID- 3039669 TI - Epstein-Barr virus: the spectrum of its manifestations in human beings. AB - Epstein-Barr virus (EBV) infects virtually everyone by adulthood, and a lifelong latency is maintained. It infects children silently, whereas the majority of adolescents have infectious mononucleosis (IM). Children who have IM before 5 years of age are often heterophil negative; EBV-specific antibodies are required for diagnosis. On rare occasions the symptoms of IM may persist in a chronic or recurrent form, and fatal infectious mononucleosis occurs rarely. Depending on the type and degree of immune deficiency and the time the EBV infection occurs in the life cycle, various atypical outcomes can occur. Children with primary immune deficiency can have fatal or chronic IM, malignant B cell lymphoma, virus associated hemophagocytic syndrome, aplastic anemia, or acquired hypogammaglobulinemia. The various outcomes of the EBV infections are likely governed by the immune response of the individual. The increased frequency of B cell neoplasms in immunodeficient patients is likely due, in part, to EBV. Individuals with acquired immune deficiency disorders such as AIDS or allograft recipients may develop malignant B cell lymphomas which tend to be polyclonal, but which may progress through stages of oligoclonality to monoclonality. This conversion likely results from specific reciprocal chromosomal translocations such as t(8;14), which is seen in Burkitt's lymphoma. Detection of EBV in immunodeficient patients is achieved by EBV-specific antibody studies or isolation of virus by obtaining spontaneous lymphoblastoid cell lines from peripheral blood, isolating virus from throat washings, or identifying EBV genome by molecular hybridization techniques. Prevention of primary immune deficiency by early detection and genetic counseling and monitoring of patients for occurrence of EBV infection may lead to early treatment. Acyclovir and immunoglobulin therapy can be of value in some patients with active EBV infection. PMID- 3039670 TI - Prospective comparative study of intradiscal high-dose and low-dose collagenase versus chymopapain. AB - The mixed results of two studies on intradiscal therapy with collagenase versus chymopapain are presented. The first study was performed from January 1983 to March 1984 and consisted of 71 patients treated with collagenase injection (600 ABC units) and 93 patients treated with chymopapain injection (4,000 units) into lower lumbar discs. The second study was started in May 1985 and ended December 1985. The results of 41 patients injected with chymopapain and 45 patients injected with collagenase (400 ABC units) are reported. The overall success rate after 3 months was 69%/63% for chymopapain/high-dose collagenase and 73%/71% for chymopapain/low-dose collagenase and 75%/72% after 6 months for chymopapain/high dose collagenase. Eighteen percent of the chymopapain-treated patients and 21% of the collagenase-treated patients of the first study had to be operated on within 6 months and 12% of chymopapain patients and 29% of collagenase patients within 3 months in the second study. Six of the 134 patients who had chymopapain treatment had slight allergic reactions. Patients who had collagenase treatment had no allergic reactions under the same regimen of systemic prophylactic measures. Patients who had high-dose collagenase injections suffered significantly more from postinjectional pseudoradicular and low-back pain in the first 3 months. In the first study, no permanent neurologic complications occurred. Two patients in the low-dose collagenase group developed cauda equina syndromes in the 2 weeks after injection because of large extruded disc fragments. PMID- 3039671 TI - The effect of chemotherapy on the morphology of human breast carcinoma. AB - Between biopsy diagnosis and mastectomy 16 patients with carcinoma of the breast received at least three courses of chemotherapy. When the biopsies and the tumour in the mastectomy specimens were compared, an increase in nuclear size, pleomorphism, vacuolation/chromatin clumping and nucleolar prominence were seen in 14 cases. Cytoplasmic changes included accumulation of lipid. There was no correlation between these morphological changes and the disease-free survival period and they could not be used to predict response to chemotherapy. PMID- 3039672 TI - Prevalence of sexually transmitted diseases in high-risk women in the Republic of Panama. AB - This study enrolled 1,032 sexually active women attending social hygiene clinics in Panama City; clinic attendance is mandatory for women employed in houses of prostitution, bars, and cabarets. Women were interviewed, and endocervical specimens were obtained for culture of Neisseria gonorrhoeae, Chlamydia trachomatis, herpes simplex virus, and cytomegalovirus. Four occupational groups attended the social hygiene clinics: prostitutes, bar girls denying prostitution, cabaret entertainers, and streetwalkers detained by the police. Prevalence of sexually transmitted disease, nationality, race, contraceptive method, and self medication varied significantly by occupation; 31% of streetwalkers had gonorrhea as did 10% of prostitutes, 5% of bar girls, and 3% of cabaret entertainers. Rates of positive serologic tests for syphilis followed the same trend: 23% in streetwalkers, 7% in prostitutes, and 3% in nonprostitutes. Rates of chlamydial infection were significantly higher in cabaret entertainers (8%) than in any other occupational group (2%). Cytomegalovirus and herpes simplex virus infections were uncommon and were found in 5% and 1% of the women, respectively. Prevalence of N. gonorrhoeae varied with self-medication and years of "professional" experience. Only one of 160 N. gonorrhoeae isolates was resistant to penicillin and also beta-lactamase-positive. PMID- 3039673 TI - Rationale for clinical application of hyperthermia and drugs. PMID- 3039674 TI - Granular cell tumor of the orbit and ocular adnexae. AB - Although granular cell tumor (GCT) has been a distinct pathological entity since 1926, confusion and controversy have long existed regarding the nature and histogenesis of the tumor. Twenty-five cases of granular cell tumor involving the orbit and ocular adnexae previously reported in the ophthalmic literature, are reviewed and six additional cases are reported. The histopathological and immunohistochemical characteristics of granular cell tumors in this uncommon location are discussed and their probable origin from Schwann cells is also considered. PMID- 3039675 TI - The primary localization of free radical generation after anoxia/reoxygenation in isolated endothelial cells. AB - While free radical-mediated reperfusion injury is clearly important in a variety of disparate organs, the particular cellular source of these radicals is unclear. To address this question, we subjected relatively pure (92% +/- 3% by factor VIII immunoassay) cultures of rat pulmonary artery endothelial cells to 0 to 45 minutes of anoxia (95% N2, 5% CO2), followed by reoxygenation (95% air, 5% CO2), to simulate ischemia/reperfusion. Cell injury was assayed after reoxygenation by the release of previously incorporated 51chromium and/or lactate dehydrogenase, and viability was determined by means of trypan blue exclusion. These three end points correlated closely. Without anoxia, the cells remained viable, with minimal evidence of injury for the entire experimental period, while 45 minutes of hypoxia followed by 30 minutes of reoxygenation produced substantial evidence of cell injury in 71% +/- 6% of the cells. This injury was reduced to 21% +/- 2% by treatment with the highly specific free radical scavengers superoxide dismutase and catalase together, either before anoxia or after anoxia, but just before reoxygenation. Similar protection was provided by xanthine oxidase inhibition with allopurinol. The injury was mimicked (without anoxia) by the exogenous generation of superoxide radicals with xanthine and xanthine oxidase. These experiments establish the essential components of free radical generation at reperfusion to be localized within the isolated endothelial cell in the absence of neutrophils or parenchymal cells. PMID- 3039676 TI - Lipoxygenase pathway inhibition impairs the allograft response. AB - The inflammatory response may play a critical role in determining the ultimate fate of the allograft. Certain inflammatory mediators derived from the arachidonic acid lipoxygenase pathway (e.g., leukotriene B4), stimulate T cell function in vitro, but their role in allograft rejection is unknown. Nordihydroguaiaretic acid (NDGA) inhibits both the lipoxygenase (LO) pathway and T cell function in vitro. In this study, we investigated the effects of systemic LO inhibition with NDGA on the in vivo generation of allospecific and natural killer (NK) effector cells and arachidonic acid metabolites from sponge matrix allografts in mice. In control animals, generation of both allospecific cytolytic and NK cells increased progressively up to 12 days after grafting. Sponge cell synthesis of both leukotriene B4 (LTB4) and the cyclooxygenase product prostaglandin E2 (PGE2) could also be detected over this period. Recipient NDGA treatment (50 mg/kg/day) impaired both the accumulation and the specific cytotoxic potential of lymphocytes in allograft. Compared with control animals (1196 +/- 30 pg/10(6) cells), cells from recipients of NDGA produced significantly less LTB4 (55 +/- 10 pg/10(6) cells, p less than 0.01) but produced normal amounts of PGE2 (340 +/- /255 +/- 22 pg/10(6) cells, p = NS), thus proving the specificity of the NDGA treatment on LO pathway function. We conclude that cells capable of metabolizing arachidonic acid by both pathways accumulate in sponge allografts, but LO activity is specifically suppressed by systemic NDGA. Furthermore, systemic LO inhibition reduces the nonspecific inflammatory component, as well as allospecific cytolytic and NK generation, without compromising animal survival. These studies suggest that suppression of specific components of the inflammatory response associated with allograft rejection by LO inhibition may be a useful approach to selective immunosuppression. PMID- 3039677 TI - Role of cyclic nucleotides in relaxation of fetal lamb ductus arteriosus. AB - Although prostaglandin E1 is used to dilate the constricted ductus arteriosus in infants with cyanotic heart disease, the mechanism is unknown. To test the hypothesis that the cyclic nucleotides adenosine 3',5'-monophosphate (cAMP) and guanosine 3',5'-monophosphate (cGMP) play a role in relaxation, isolated rings of the ductus arteriosus of fetal lambs were studied. Tension of isometric contraction was measured by force displacement transducers. After contraction with oxygen, a control group was compared with rings in which the stimulus for relaxation was either nitrogen gas, prostaglandin E1 (PGE1), nitroglycerin (NTG), or nitroprusside (NPS). During relaxation, tissue was frozen at 30 seconds and at 1, 2, and 5 minutes and analyzed for cAMP and cGMP. PGE1 (10(-6) mol/L) decreased tension by 33% compared with 70% for nitrogen gas, 81% for NTG (10(-5) mol/L), and 92% for NPS (10(-5) mol/L). The maximal relaxation induced by PGE1 was associated with an 11-fold increase in cAMP; PGE1 had no significant effect on cGMP tissue levels. Nitrogen gas, NTG, and NPS produced similar increases in cAMP, and eight-, 25-, and nine-fold increases in cGMP, respectively. These results suggest that the patency of the ductus arteriosus is dependent on activation of both guanylate cyclase and adenylate cyclase and that the nitrovasodilators may be clinically useful in maintaining patency of the ductus arteriosus. PMID- 3039678 TI - Intracellular pH (pHi) in gastric surface epithelium is more susceptible to serosal than mucosal acidification. AB - Intracellular microelectrode techniques were used to examine the effects of mucosal or serosal acidification on intracellular pH (pHi) in gastric surface epithelial cells. Necturus antrum was mounted in a modified Ussing chamber, and pHi was determined from the difference between the potentials recorded by intracellular conventional and pH-sensitive microelectrodes. In tissues bathed with bicarbonate-buffered Ringer's solution (pH 7), acidification of the mucosal solution to pH 4.5 by isotonic replacement of the NaHCO3 with NaCl had no significant effects on pHi. In contrast, acidification of the serosal solution to pH 4.5 by replacing the bicarbonate reduced pHi from 7.32 +/- 0.04 to 6.95 +/- 0.06 (p less than 0.001, n = 8). Similarly, in tissues bathed with HEPES-buffered Ringer's solution (pH 7.0), pHi was unaffected by reducing the mucosal solution pH to 4.5 with HCl but fell 0.21 +/- 0.05 pH units (p less than 0.01, n = 7) during acidification of the serosal solution to pH 6. These results suggest that gastric epithelium is more sensitive to acidification from the serosal than the mucosal side. Such a finding is consistent with the concept of a gastric mucosal barrier to luminal acid. It may also explain the gastric epithelium's greater sensitivity to acute ulceration during systemic acidosis. PMID- 3039679 TI - Inhibition of established rat fibrosarcoma growth by the glucose antagonist 2 deoxy-D-glucose. AB - Sarcoma cells exhibit higher rates of glycolysis than normal tissues and may be dependent on glucose utilization for growth. Accordingly, we tested the ability of the glucose antimetabolite 2-deoxy-D-glucose (2-DG) to inhibit the growth of an established methylcholanthrene-induced rat fibrosarcoma in three groups of F344 rats with increasing subcutaneous inoculations of tumor (2 X 10(6) cells, 1 X 10(7) cells, and 1 mm tumor fragments). Rats were randomized to receive 2-DG or saline solution at doses of 0.75 gm/kg, 1.5 gm/kg, or 1.75 gm/kg, beginning 3 days after tumor implantation and continuing for 10 days. Tumors were removed and weighed on day 14. We measured tissue [14C]-2-DG levels in tumor, brain, liver, and muscle after intraperitoneal injection of radiolabeled 2-DG. In these same tissues we determined the activity of glucose-6-phosphatase (G-6-Pase), an enzyme which dephosphorylates the intracellular glycolytic inhibitor 2-DG-6-phosphate, thus reversing the antitumor effect of 2-DG. All groups treated with 2-DG had a significant reduction in tumor weight of 50% to 70% when compared with saline solution-treated controls. Toxicity was substantial at the highest dose of 2-DG, but minimal toxicity was noted at intermediate and low doses. Tumor had the greatest uptake of [14C]-2-DG, with low levels of G-6-Pase leading to prolonged retention and highest tissue levels of radiolabeled 2-DG. Use of 2-DG inhibits established sarcoma growth because it is rapidly transported into tumors, cannot be metabolized after phosphorylation, and is dephosphorylated and released slowly from tumor cells. Rat sarcoma growth is dependent on glucose utilization and can be effectively inhibited by glucose antagonism. PMID- 3039680 TI - [Dental pain: physiology and anatomy]. PMID- 3039681 TI - Oral human papillomavirus infections. PMID- 3039682 TI - Suppression of host defences by Aspergillus fumigatus. AB - An important feature of the microbicidal action of phagocytic cells is their ability to produce reactive oxygen intermediates. In an attempt to identify the mechanisms by which the fungus Aspergillus fumigatus resists normal host defences the effect of spores and spore diffusates of A fumigatus on the production of superoxide anion and hydrogen peroxide by primed rodent phagocytic cells has been measured. For comparison we have used the non-pathogenic fungus Penicillium ochrochloron. Production of these reactive oxygen intermediates in response to A fumigatus was significantly lower than that in response to P ochrochloron. A similar reduction was achieved by diffusate prepared from freshly washed spores. The inhibitory component was of low molecular weight (less than 14,000) and its effect was dose dependent. These results suggest that spores of A fumigatus fail to trigger and also inhibit the production of reactive oxygen intermediates by phagocytic cells. PMID- 3039683 TI - Treatment of small cell lung cancer by pneumonectomy and single course high dose chemotherapy. PMID- 3039684 TI - Interference of the thromboxane antagonist SK&F 88046 with platelet activation and subsequent desensitization by arachidonic acid and the thromboxane mimetics U46619 and EP171. AB - We have investigated the effects of the thromboxane antagonist SK&F 88046 on human platelet activation and desensitization by arachidonic acid (AA) and by the thromboxane A2 mimetics U46619 or EP171. SK&F 88046 inhibited platelet aggregation and secretion induced by AA, U46619, EP171 and thrombin at low (0.05 U/ml) but not at a high (1 U/ml) concentrations. Platelet inhibition was reversed by washing the cells. Platelets pre-exposed to AA, U46619 or EP171 and then disaggregated with prostacyclin, washed and resuspended, failed to respond with aggregation or secretion to a second challenge by either agonist. In the presence of the endoperoxide/thromboxane receptor antagonist L636499 or of SK&F 88046, pre exposure to AA, U46619 or EP171 failed to prevent subsequent responses to the related agonists. Our results suggest that SK&F 88046 is a selective antagonist of platelet activation and desensitization induced by TxA2 or prostaglandin endoperoxides and that it may have utility as an anti-platelet drug. PMID- 3039685 TI - [Foot and mouth disease--then, now and in the future]. AB - Foot-and-mouth disease was a problem as early as 1862; however, insights into the aetiology and particularly the concept of infectious disease had not yet been elaborated. Our knowledge and the possibilities of dealing with the disease have shown a spectacular increase since then. Foot-and-mouth disease is now under control in large parts of Europe; this is the result of strict measures and mass vaccination of cattle. However, the situation is unstable, as is testified to by the situation in Italy, and there are questions regarding the origin of sporadic cases of the disease. The main question is: how do we reach the ultimate objective: a Western Europe free from foot-and-mouth disease, and where annual vaccination of cattle no longer is applied? The problems are analysed. An international approach appears to be essential. PMID- 3039686 TI - [Feline leukemia virus (FeLV) and FeLV-associated diseases in cats: a review]. AB - Feline leukaemia virus (FeLV) usually occurs in its natural species, the domestic cat. FeLV is also important to human individuals as a comparative model, as FeLV may cause a variety of diseases which are partly malignant and partly benign, such as immunosuppression which bears a resemblance to AIDS (acquired immune deficiency syndrome) in man. Although FeLV is a common infective agent, the incidence of disease due to FeLV is much higher in cats kept in closed households in which several of them are present than it is in free-range cats. Consequently, diseases caused by FeLV are frequently diagnosed in pedigree cats which are often maintained in relatively large numbers. FeLV is transmitted among cats by contagion. The main sources of infection are persistantly infected FeLV carrier cats which continuously excrete virus from the mouth and in other secretions. The majority of cats infected with FeLV will produce neutralising antibodies. Cats which are unable to do so, will become permanently infected. The prognosis is bad in these cats: 90 per cent will die within five years. Various techniques are used to detect FeLV. The most common method, the indirect immunofluorescence (IFA) test, is performed on air-dried blood smears. The results of the IFA agree with that are almost completely identical to those of the virus isolation test. Another test is ELISA (enzyme-linked immunosorbent assay), which produces approximately 10 per cent more positive results which are probably due to circulating FeLV antigens. Dissemination of FeLV among cats may be prevented by identifying infected carrier cats and removing them from contact with non infected cats. Removal programmes using indirect immunofluorescent antibody (IFA) tests were used successfully in the Netherlands. The proportion of FeLV-positive cats decreased from 9 per cent in 1974 to approximately 3 per cent in 1985 during such a removal programme. During the above period, the removal programme was carried out in the society of Dutch cat breeders 'Felikat', the programme being made compulsory on all members of the society. The incidence of cats positive for FeLV decreased from over 11 per cent in 1974 to less than 2 per cent within four years. None of the cats tested in this society was found to be positive for FeLV in 1984 and 1985. Besides removal programmes, other methods of control, such as vaccination, were developed to prevent the spread of FeLV. The FeLV immunostimulating complex vaccine (FeLV-ISCOM vaccine) a subunit vaccine in which FeLV-gp70 is presented in a particular manner, seems to be promising. PMID- 3039687 TI - [Immunization schedule for parvovirus infection?]. PMID- 3039688 TI - [Infections with feline leukemia virus (FeLV) in postmortem cats]. AB - Cats presented for post-mortem examination were examined by immune histological techniques for the presence of persistent FeLV infections. Persistent FeLV infection was found to be the most common fatal infectious disease currently occurring in cats. It was detected in 16 per cent of the cats referred for post mortem examination, whereas a proportion of 3 per cent may be regarded as likely in the normal population. PMID- 3039689 TI - Recombinant gamma interferon induces class II major histocompatibility complex antigens on insulinoma cells. AB - Aberrant expression of major histocompatibility (MHC) antigens has been implicated as a factor contributing to organ-specific autoimmunity, such as progressive loss of pancreatic beta cells in type 1 diabetes. We investigated the potential of a rat beta cell tumour, RINM5F, to express enhanced levels of MHC antigens in vitro. To this purpose we treated RINM5F cells in vitro with recombinant rat gamma interferon (rIF gamma). We used monoclonal antibodies to RT1.A (class I) and RT1.B (class II) antigens of the rat MHC in conjunction with flowcytometry and immunoperoxidase techniques to analyse the expression of MHC antigens. Untreated RINM5F cells express low levels of RT1.A, whereas they are negative for RT1.B. Treatment with rIF gamma appeared to increase the expression of RT1.A antigens substantially. Most importantly, RT1.B antigens were newly expressed by rat insulinoma cells in vitro after treatment with rIF gamma. To our knowledge this is the first documentation of the potential of beta cells or their derivatives to express class II MHC antigens following IF gamma-treatment. This mechanism may play an important role in the augmentation and perpetuation of insulitis leading to type 1 diabetes mellitus. PMID- 3039690 TI - [The clinical administration of the opioid antagonist naloxone in dogs]. AB - The short introduction gives a review on the complex of exogen and endogen opioids and their receptors as well as on their pharmacodynamics, pharmacokinetics and toxicity of naloxone. The clinical efficacy of naloxone as an opioid antagonist is described. Applications of naloxone for the dog are specified: antagonisation of etorphine, morphine, levomethadone and fentanyl, antagonisation of exogen and endogen opioids in puppies and treatment of lactomania in the bitch. The mean effective dose to antagonize morphines is 0.003 mg/kg bodyweight. If persisting analgesia is indicated the dose of naloxone in titrating steps in 0.001 mg/kg bw. To antagonize postpartal hypoxia in puppies 0.02 mg per animal naloxone have to be injected. For treatment of lactomania a dose of 0.01 mg/kg bodyweight twice a day for a couple of days is recommended. The clinical effectivity of naloxone is proved doubtlessly. Compatibility and safety are very high. PMID- 3039691 TI - Seroconversion from HBs-Ag to anti-HBs in a case of liver cirrhosis associated with hepatocellular carcinoma. AB - This paper reports a case of liver cirrhosis associated with hepatocellular carcinoma (HCC) of a woman who was converted from hepatitis B surface antigen (HBs-Ag) positive to antibody against HBs-Ag (anti-HBs) positive in the serum through an immunoregulatory steroid rebound phenomenon. The histology of the biopsy specimen taken before the seroconversion showed an early stage of liver cirrhosis with moderate infiltration of mononuclear cells. At autopsy about 3 years after the seroconversion, the liver tissue free of the tumor was in an early stage of liver cirrhosis. Fibrosis did not advance as compared with the biopsy specimen. In addition, mononuclear cell infiltration decreased remarkably and piecemeal necrosis disappeared after the seroconversion. The immunohistologic examination of hepatocytes demonstrated that positive stainings for HBs-Ag and for hepatitis B core antigen (HBc-Ag) in the biopsy specimen turned to be negative in the autopsy specimen. These facts indicate that the steroid rebound phenomenon eliminated free hepatitis B virus (HBV) in the hepatocytes in the absence of massive necrosis of hepatocytes. HBV-DNA integration was proved in the genome of HCC by molecular hybridization method. PMID- 3039692 TI - Effects of phospholipases C from bacteria on binding of enkephalin to rat brain membranes. AB - The effects of phospholipases C on the equilibrium constants and maximal binding capacities of tritiated [D-Ala2,-D-Leu5] enkephalin to rat brain membranes were investigated using phosphatidylcholine-hydrolyzing phospholipase C and sphingomyelinase C of Bacillus cereus and, phosphatidylinositol-specific phospholipase C of Bacillus thuringiensis. When 72% of the phosphatidylinositol in the rat brain membranes was hydrolyzed by phosphatidylinositol-specific phospholipase C, the affinity of opiate receptor for [D-Ala2,-D-Leu5] enkephalin was almost doubled and maximal binding of [D-Ala2,-D-Leu5] enkephalin was decreased to 87% of control. Although specific [D-Ala2,-D-Leu5] enkephalin binding was decreased with phosphatidylinositol hydrolysis when measured at higher concentration (30 nM) of [D-Ala2,-D-Leu5] enkephalin, the specific binding was increased with the hydrolysis of phosphatidylinositol when measured at lower concentration (6 nM) of the ligand. On treatment of membranes with phosphatidylcholine-hydrolyzing phospholipase C, specific [D-Ala2,-D-Leu5] enkephalin binding was drastically decreased with the progressive hydrolysis of phosphatidylcholine in the rat brain membranes, and specific binding was completely lost after 81% hydrolysis of phosphatidylcholine. However, the affinity of opiate receptor for [D-Ala2,-D-Leu5] enkephalin was not influenced, and maximal binding was decreased to 32% of the control when 61% of phosphatidylcholine was hydrolyzed. Treatment with sphingomyelinase C did not cause any appreciable reduction of specific [D-Ala2,-D-Leu5] enkephalin binding. From these results, it is concluded that the binding of [D-Ala2,-D-Leu5] enkephalin to opiate receptor is influenced by changes in the phospholipid environment of the rat brain membranes, and that phosphatidylinositol may be a modulator for the function of the receptor. PMID- 3039693 TI - Characteristics of opioid receptors in guinea-pig cerebellum and human placenta. AB - Characteristics of opioid receptors were examined in guinea-pig cerebellum and human placental membrane preparations. Two binding sites for [3H] ethylketocyclazocine ([3H]-EKCZ) were found in guinea-pig cerebellum membranes whereas only one binding site was detected in human placental membranes. The potencies of kappa opioid agonists were decreased in competing with the binding of [3H]-naloxone to guinea-pig cerebellum membranes or human placental membranes by 5'-guanylylimidodiphosphate (Gpp(NH)P) and sodium. These results suggest that there may be true kappa receptors in human placenta and that kappa receptors in both preparations may couple with the adenylate cyclase system. PMID- 3039694 TI - Capsular formation surrounding hepatocellular carcinoma. AB - In Japan, it is well known that most cases of hepatocellular carcinoma occur in the cirrhotic liver and this fact is regarded as a pathologic characteristic of hepatocellular carcinoma. Capsule formation surrounding the tumor is also a characteristic of hepatocellular carcinoma and it may be related to the developmental mode of hepatocellular carcinoma, to the clinical manifestation of symptoms and to the prognosis of the disease. The formation of the capsule can often be detected by means of modern imaging diagnostic procedures as well as sonography-guided or laparoscopy-guided needle biopsy of the liver. This paper reports the results of an investigation on the incidence of capsular formation surrounding hepatocellular carcinoma and its clinicopathological consideration in hepatocellular carcinoma in its early stage occurring in liver cirrhosis. PMID- 3039695 TI - Murine cytomegalovirus infection model in Balb/c mice--1. Virological and pathological profiles in mice inoculated with various virus doses. AB - By intraperitoneal injection of murine cytomegalovirus (MCMV) in Balb/c mice at doses of 2 X 10(6), 2 X 10(5) and 2 X 10(4) pfu per mouse, lethal, acute but non lethal and asymptomatic infection were produced respectively. 100% mortality was found in mice infected with the lethal dose, as well as high MCMV titres in liver and spleen, severe necrosis of the spleen and severe degeneration of the liver. In these mice leukocyte counts in blood was not raised and inflammatory responses in the peritoneum, spleen and liver were not observed. Mice with acute but non lethal infection showed less severe pathological changes, moderate spleen necrosis was followed by spleen enlargement. In this group of mice, severe mononucleosis was observed, and moderate inflammatory reactions were noted in the peritoneum, liver and spleen. Mice asymptomatically infected exhibited slight mononucleosis in the blood and splenomegaly. PMID- 3039696 TI - Murine cytomegalovirus infection model in Balb/c mice--2. Pericarditis with myocardial involvement during virus infection. AB - Pericarditis with myocardial involvement was observed in mice infected with doses of murine cytomegalovirus (MCMV) designed to produce lethal, acute but non-lethal and asymptomatic infections. White spot was noted on the surface of the right ventricle from 4 to 6 days after MCMV infection. MCMV was isolated from the heart tissues as early as 2 days after infection. The virus titre reached a peak on day 6 and thereafter rapidly declined to undetectable level. Histopathological changes appeared on day 2 to 4 when necrosis of myocardial fibres occurred in the subpericardial region. This was followed by acute inflammatory cellular infiltration on day 6. After that organization of fibrin took place and fibrotic lesion and cellular infiltration could still be observed after 18 days. The pathological changes could only be seen in the right ventricle. Peroxidase labelled antibody technique was carried out to detect MCMV antigen in the cardiac tissue. MCMV antigen was found in the pericardium only at the early stage of infection. PMID- 3039697 TI - A dehydroepiandrosterone sulfatase inhibitory activity in soluble proteins of guinea pig liver. AB - An inhibitor of microsomal dehydroepiandrosterone sulfatase was found in the soluble fraction of non-pregnant guinea pig liver. The extent of inhibitory effect was dependent on the concentration of soluble proteins. The inhibitor was partly purified by gel permeation and hydroxylapatite chromatography with a purification factor of 16.6. The soluble inhibitor was non-dialyzable, not destroyed by RNase or DNase digestion but totally destroyed by pronase digestion. The inhibitor is a soluble protein with a molecular weight of approximately 17,000 (determined by gel permeation chromatography). Inhibition of microsomal dehydroepiandrosterone sulfatase by the soluble inhibitor is a non-competitive inhibition. From this present finding the question arises whether the inhibitor could be involved in the regulation of the hydrolysis of dehydroepiandrosterone sulfate in the guinea pig liver. PMID- 3039698 TI - Liver transplantation in children: the Hannover experience. PMID- 3039699 TI - Pediatric liver transplantation: patient evaluation and selection, infectious complications, and life-style after transplantation. AB - Liver transplantation is an increasingly accepted treatment for children with end stage liver disease. Evaluation of the patient and appropriate patient selection for transplantation will become increasingly important issues as more and more children come to transplantation and compete for available organs. Numerous complications occur after transplantation, including infections. We have summarized our experience with bacterial, fungal, and viral infections in these patients and emphasize the need for continued improvement in immune suppressive drugs and regimens to minimize such complications. And finally, information presented on 65 pediatric patients followed 2 to 5 years suggests that, despite numerous complications and often prolonged hospitalization for transplantation, life-style after transplantation appears to be significantly improved. PMID- 3039700 TI - Results of prospective matching for cytomegalovirus status in renal transplant recipients. PMID- 3039701 TI - Failure to preserve liver allografts for 24 hours: experimental and theoretical considerations. PMID- 3039702 TI - Atherosclerosis: diet and risk factors. PMID- 3039703 TI - [Effect of a number of biologically active substances on the activity of membrane bound Na+,K+-ATPase from the bovine brain]. AB - Cellular Na+, K+-transport ATPase appears to be more resistant to the influence of DNA intercalators and other agents than ATP-synthetase complex of inner mitochondrial membrane. Neither synthetic intercalators, nor antibiotics olivomycin and actinomycin D, some uncouplers of oxidative phosphorylation, alkaloids and inorganic bivalent cations selectively (up to 10(-4) M) inhibit Na+, K+-ATPase activity. Besides ouabain, alkaloid sanguinarine and cation Ag+ (the latter in the absence of anion Cl-) cause a significant decrease of Na+, K+ ATPase activity. Ag+ is most potent as inhibitor, while sanguinarine is weaker than ouabain. Inhibition of Na+, K+-ATPase by sanguinarine may result presumably from modification of SH-groups of the enzyme. PMID- 3039705 TI - Unusual crystalline inclusions in a case of AIDS-related complex. AB - A lymph node of a patient with AIDS-related complex was studied with light and electron microscopic techniques. In numerous plasma cells and follicular center cells amorphous and unusual crystalline structures which probably consist of abnormal immunoglobulin could be found. PMID- 3039704 TI - Electron microscopy in the diagnosis of percutaneous fine needle aspiration specimens. AB - Five years experience in the application of electron microscopy to fine needle aspiration biopsy specimens is reviewed. In an initial evaluation, 200 consecutive unselected specimens were examined; 89 proved diagnostic and, in a third of these, electron microscopy gave additional information that was often essential to diagnosis. Negative specimens resulted almost entirely from failure to obtain an adequate amount of material. Results were improved by the adoption of a preparation technique involving concentration of the cells in bovine serum albumin and by the inspection of a rapidly stained smear at the time of the aspiration procedure, with a further needle pass for electron microscopy being performed if necessary. Despite the small size of specimens, adequate examination was usually possible and electron microscopy has proved of value in the diagnosis of tumor samples acquired by fine needle aspiration in the same way as has been established with the larger sized specimens obtained by conventional biopsy and surgical resection. PMID- 3039706 TI - Intracytoplasmic crystals in endothelial cells. PMID- 3039707 TI - Familial occurrence of testicular cancer. AB - A case of testicular cancer in 2 brothers is reported and a review of the literature about testicular malignancies and etiologic factors in closely related family members is given. The familial incidence of testicular tumor is found to be 3% in twins and 0.6-2.1% in less closely related men. Tumors were of the same histology in 70-77% of the twins, whereas in brothers and in other degrees of relationship tumors of different histology mostly occur. Following the diagnosis of the tumor in the first man, the average interval to presentation of the tumor in the relative was 3.7 years in twins, 7.7 years in nontwin brothers and 13.5 years for less closely related men. The need for a thorough checkup of other family members is advised. PMID- 3039708 TI - Malignant fibrous histiocytoma arising from the renal capsule: report of a case. AB - We report a case of malignant fibrous histiocytoma arising from the renal capsule. The tumor was found during screening ultrasonography. Surgical excision and adjuvant chemotherapy consisting of cyclophosphamide, vincristine, adriamycin and actinomycin D were performed. There has been no recurrence as of 2 years and 4 months after the operation. We reviewed the 5 other cases of malignant fibrous histiocytoma arising from the renal capsule. PMID- 3039709 TI - Abnormalities of ejaculation. AB - The normal physiologic processes of emission and ejaculation require coordination of neurophysiologic, anatomic, and, in certain cases, psychological phenomena. Disruption of any component, from the embryologic development of the mullerian duct through the medications used for nonrelated systemic disease, can alter the efficient function of ejaculation. Evaluation of the urologic patient who has any of a number of abnormal ejaculatory states requires an understanding of the many possible mechanisms of failure. The majority of these men need evaluation because of a possible male-factor infertile marriage. The potential for improvement is significant, given the development of improved techniques to stimulate ejaculation and the promise shown by extracorporeal fertilization techniques such as in vitro fertilization and gamete intrafallopian tube transfer. These patients deserve complete assessment and optimization of any factors that will enable them to achieve their goal of procreation. PMID- 3039710 TI - Triple therapy for adult Wilms tumor. AB - Adult Wilms tumor is still considered a rarity. Approximately 170 adult Wilms tumors have been reported. The final diagnosis is usually established with surgery. There is no specific radiologic diagnosis of adult Wilms tumor. We report on a case of Wilms tumor in a twenty-two-year-old black man. The tumor was removed with radical nephrectomy and classified as Stage II. Radiation of the renal fossa (4,000 rad) and chemotherapy with actinomycin D and vincristine were administered after surgery. Six years after surgery the patient is disease free. A review of published cases of adult Wilms tumor is presented, and a plea is made for triple therapy. PMID- 3039712 TI - Evidence of hepatitis A infection in immature rhesus monkeys. AB - Forty immature (less than 2 years old) rhesus monkeys (Macaca mulatta) with marked increases in aspartate and alanine aminotransferase activities were examined. Serological and histopathological evaluations were done to determine if affected animals were infected with hepatitis A virus. Although no clinical signs of illness were noted in any of the monkeys, an excellent correlation was found between the increased serum aminotransferase values and seropositivity with the acute phase (IgM) HAVAB-M antibody. Histopathological evaluations of livers of selected animals revealed hepatic lesions consistent with those in chimpanzees and marmosets infected with hepatitis A virus: generalized activation of sinusoidal lining cells, focal hepatocellular necrosis with occasional acidophilic bodies, and cuffs of mononuclear cells in the portal areas. Although no animals were seropositive for HAVAB upon receipt from the breeding colony, a total of ten of 18 animals for which serological data were available had seroconverted to HAVAB positivity during the 4-month observation period. These results are consistent with hepatitis A infection in immature rhesus monkeys and indicate the potential significance of serological testing in animals in which hepatic function is being evaluated. PMID- 3039711 TI - Thyroid hormone and Na,K transport. PMID- 3039713 TI - Concurrent malignant catarrhal fever and bovine virus diarrhoea virus infection in a dairy herd. AB - An account is given of an outbreak of malignant catarrhal fever which occurred in a 98-cow dairy herd. Ten animals died or were slaughtered and the disease was confirmed by clinical and histological examination. Serological tests for malignant catarrhal fever virus were positive in three of four animals. The diagnosis of malignant catarrhal fever was complicated by the presence of bovine virus diarrhoea virus infection in three of the early cases. The initial cases of malignant catarrhal fever occurred in a group of nine-month-old calves which were housed in an old milking parlour with 19 pedigree Suffolk ewes at lambing time. Later cases occurred in two adult cows and in two heifers. Investigations of the remainder of the herd for evidence of bovine virus diarrhoea virus did not reveal the presence of any persistently infected cattle. Serological examinations for antibody to malignant catarrhal fever and bovine virus diarrhoea virus were carried out on the 19 Suffolk ewes. Six of them had neutralising antibody titres to malignant catarrhal fever virus and three were positive in the indirect immunofluorescence test. The possible roles of bovine virus PMID- 3039714 TI - Immunoblastic lymphoma and cholangiocarcinoma in a cat. PMID- 3039715 TI - Deaths in swans and geese. PMID- 3039716 TI - Actions of BW540C in an equine model of acute inflammation: a preliminary study. AB - An equine model of acute non-immune inflammation has been developed to facilitate studies of the inflammatory process and the actions of novel anti-inflammatory drugs. Five polyester sponge strips soaked in sterile 2% carrageenin solution were placed in subcutaneous pouches prepared under local anaesthesia in the necks of conscious ponies. Serial removal of the strips and harvesting of the exudate enabled studies to be made of the cellular, biochemical and mediator aspects of the localised, acute inflammation, and the heat generated by the lesion was monitored by infra-red thermometry. Maximal concentrations of the eicosanoids 6 keto-prostaglandin F1 alpha, thromboxane B2 and leukotriene B4 occurred at 9 h, whereas leukocyte numbers, lactate dehydrogenase (LDH) and total protein concentrations were greatest at 24 h. Lesional skin temperature was increased by approximately 4 degrees C throughout the 24 h period. The novel anti-inflammatory agent BW540C, administered orally at a dose-rate of 20 mg/kg, did not affect leukocyte infiltration or the concentrations of protein, LDH and eicosanoids in exudate but serum thromboxane B2 levels were reduced. Skin temperature rises were greater in drug-treated animals. It is concluded that higher doses of BW540C will be required for a clinically useful anti-inflammatory action in horses. PMID- 3039717 TI - [Evaluation of the extent of central lung cancer based on x-ray, radiologic and endoscopic data]. PMID- 3039718 TI - Immunobiology of bluetongue virus. AB - Following BTV infection or vaccination, sheep develop both anti-virus antibody (which may include neutralizing antibody) and a cellular immune response. Yet, it still is unclear what aspects of the response are most critical in preventing infection and disease from this virus. This is, in part, the result of a lack of knowledge of all of the viral virulence factors. However, with the current work involving subunit vaccines and the efforts to more carefully characterize virulence factors and tissue tropism, there will be a more thorough understanding of the immunobiology of the Bluetongue virus. PMID- 3039719 TI - Pathogenesis of visna/maedi and caprine arthritis-encephalitis: new leads on the mechanism of restricted virus replication and persistent inflammation. AB - Lentiviruses are unique retroviruses which cause diseases with long incubation periods and prolonged clinical courses. The prototype lentiviruses, visna/maedi of sheep and arthritis-encephalitis virus of goats (CAEV), infect cells of the monocyte-macrophage system and replicate at a restricted level in these cells. The virus life cycle is closely associated with maturational factors in the cells; monocytes support the early stages of the replication cycle which goes to completion only when the cells mature to macrophages. Virus replication in the monocyte-macrophage results in lesions characterized by mononuclear cell infiltration of the central nervous system (CNS), lungs, synovium and mammary gland and their draining lymph nodes. Co-cultivation of sheep or goat lymphocytes with macrophages infected with visna or CAE viruses results in production of a unique interferon (LV-IFN). LV-IFN is a non-glycosylated protein of 54,000 to 64,000 daltons and has biological properties which have several implications for pathogenesis. Firstly, it retards the rate of maturation of monocytes and thus indirectly slows the rate of virus replication. Second, it restricts the rate of virus replication in mature macrophages by preventing virus maturation. Third, it induces expression of class II (Ia) antigens of the major histocompatibility complex on cells of macrophage lineage. Thus, by curtailing virus replication and enhancing expression of MHC class II antigens, LV-IFN may contribute to the induction and augmentation of the host's lymphoproliferative response to the virus. PMID- 3039720 TI - [Demonstration of a reovirus in helicopter disease of broilers]. AB - A reovirus was isolated from 5-13-day-old broiler birds with signs of the so called helicopter disease--one of the forms of the malabsorption syndrome. A number of organs were investigated (trachea, liver, spleen, bursa of Fabricius, proventricular stomach, and tendon sheath of musculus gastrocnemius). The virus was isolated from the proventricular stomach and the tendon sheath in cell cultures of chick embryo kidney as well as in 5-7-day-old chick embryos inoculated in the yolk sac. The remaining organs proved virologically negative. All organs, including the primarily positive ones, were shown to be virologically negative upon the inoculation of chick embryo fibroblast cultures and of chorioallantoic membranes of 10-11-day-old chick embryos. The isolate was identified as an avian reovirus, and was denoted with the initials R-85--on the grounds of its biologic and physico-chemical properties, of its morphology (negative-contrast electron microscopy), and of its morphogenesis (ultrathin electron-microscopic cross sections). Investigated were the histopathologic changes in chick embryos inoculated with the R-85 reovirus and the pathologic changes in the liver of spontaneously affected birds. PMID- 3039721 TI - Frequency distribution of plasma cells in the medullary cords of tumour-draining axillary and paracolic lymph nodes. AB - Five hundred and ninety-seven axillary lymph nodes draining 104 invasive ductal breast cancers, and 94 paracolic lymph nodes draining 30 invasive adenocarcinomas of the large bowel were investigated immunohistologically to determine the frequency distribution of plasma cells (PC) in the medullary cords (MC). The degree of plasmacytic infiltration was calculated semiquantitatively using the 3 grade scale (0/+, + +, + + +) of Cottier et al. (1973). Statistical analysis yielded the following results: While a marked reactive plasmacytosis (+ + +) was seen in 28.7% of the paracolic lymph nodes, only 1.5% of the axillary lymph nodes exhibited a comparable degree of plasmacytic infiltration (p less than 0.0001). Conversely, low PC counts (0/+) were encountered in 51.1% of the paracolic lymph nodes, but in 83.9% of the axillary lymph nodes. A comparison of axillary lymph nodes with and without nodal metastasation revealed no significant differences (nodal-negative cases: 0/+: 83.6%, + +: 14.3%, + + +: 2.1%; nodal-positive cases: 0/+: 84.3%, + +: 14.9%, + + +: 0.8%). However, significantly more (p less tha 0.001) paracolic lymph nodes of the nodal-negative group revealed a marked plasmacytosis, whereas in the nodal-positive group lymph nodes with low PC counts were more frequent (nodal-negative cases: 0/+: 27.7%, + +: 29.7%, + + +: 42.6%; nodal-positive cases: 0/+ 74.5%, + +: 10.6%, + + +: 14.9%). The degree of plasmacytic reactions in the tumour-regional lymph nodes was not related to the stage of the primary tumour. Moreover, no correlation exists between the PC content of the MC and the amount of PC in metastatic deposits of the same lymph nodes. Altogether, these results do not support the concept that the plasmacytic reactions in the MC of tumour-draining lymph nodes are chiefly determined by effects (stimulating or suppressing) of the primary carcinomas. The topography of the lymph nodes, however, seems to be the main determinant influencing the PC content of MC. PMID- 3039722 TI - Ectopic anterior pituitary corticotropic tumour in a six-year-old boy. Histological, ultrastructural and immunocytochemical study. AB - The report documents a silent, oncocytic, ACTH-producing ectopic anterior pituitary tumour in a 6-year-old boy. The invasive intrahemispheric neoplasm had no connection with the pituitary gland, the sella turcica or the sphenoid sinuses. The apparent similarities existing between this tumour, some choroid plexus carcinomas and steroid-producing neoplasms are discussed. PMID- 3039723 TI - [Circulation of parainfluenza viruses and adenoviruses in groups exposed to the action of noxious chemicals or not]. AB - Investigations were conducted during 1985 and 1986 years on the effect of some chemicals on the parainfluenza and adenovirus circulation in an industrial enterprise community. The presence of type 1, 2 and 3 parainfluenza virus and of adenovirus was revealed by immunofluorescence in exfoliated cells collected from nasopharynx. The kinetic of specific hemagglutination inhibiting and complement fixing antibodies was followed monthly by immunological tests. Meaning of the results is discussed from an epidemiological point of view. PMID- 3039724 TI - [Molecular mechanism of the replication of several DNA viruses]. AB - Study of the replicative mechanism of viruses is extremely important, since the knowledge of the events taking place at a molecular level can contribute to new antiviral product elaboration. A unique pattern of the DNA virus replication cannot be given, because every virus family has its own mechanism of replication. Investigations were conducted on the replication of two groups of viruses of major importance in human pathology: the hepadnaviruses and the herpesviruses, which are involved not only in acute infections, but also in some cellular transformations and even in cancerogenesis. It was shown that the replication mechanism of the hepadnaviruses has several points in common with retroviruses, fact which places them in a rather peculiar position among the DNA viruses. On the other hand, the herpes viruses have their own characteristics such as the four genome isomeres, whose significance is yet unclear. The viral message transcription and translation stages, as well as the mechanism of replication of the genome are described. PMID- 3039725 TI - [Risk of transmission of enterovirus from the surface of toilets]. AB - Experimental studies using dyes and microbial agents as indicators showed that a viral contamination of the various surfaces of the water closets is possible through the droplets formed when one flushes the W.C. pan. An investigation conducted in several public water closets allowed to reveal the presence of enteroviruses in 2.8% to 5.8% of the samples collected from various surfaces of the water closets. The results of the investigations showed that the hinged seats were contaminated the most frequently (6.8% of the samples). PMID- 3039726 TI - [Anti-proliferative and antiviral effects of human alpha-interferon on tumor cells]. AB - Antiproliferative and antiviral activities of a type alpha human leukocytic interferon on several heteroploidic cell lines: HeLa, HEp-2, and T-10, a cell line of malignant origin (glioblastoma) were investigated, as compared to subcultures of human embryo fibroblasts. The tumor cell multiplication rate decreased proportionally to the amount of interferon in the culture medium. The highest interferon concentration used in our experiments (1,000 mu/ml) induced a decrease of the normal cell multiplication rate (human embryo fibroblasts). The same amount of interferon had a cytotoxic effect against the T-10 cells, but this phenomenon is reversible if the interferon is excluded after 24 h from the culture medium. There was no quantitative relation between the magnitude of the antiviral and of the cytotoxic effects of the type alpha human interferon on the tested cellular substrates. PMID- 3039728 TI - The nucleotide sequence of dengue type 4 virus: analysis of genes coding for nonstructural proteins. AB - We recently cloned a full-length DNA copy of the dengue type 4 virus genome. Analysis of the 5' terminal nucleotide sequence suggested that the three-virion structural proteins are synthesized by proteolytic cleavage of a polyprotein precursor which is encoded in one open reading frame. We now present the remaining sequence of the dengue type 4 virus genome which codes for the nonstructural proteins. The entire genome, which is 10,644 nucleotides in length, contains one long open reading frame which codes for a single large polyprotein 3386 amino acids in length. Alignment of the dengue nonstructural protein sequence with that of other flaviviruses, including yellow fever and West Nile viruses, revealed that significant homology exists throughout the entire nonstructural region of the dengue genome and this allowed tentative assignment of individual nonstructural proteins in the following order: NS1, NS2a, NS2b, NS3, NS4a, NS4b, and NS5-COOH. Processing of the nonstructural proteins appears to involve two types of proteolytic cleavage: the first occurs after a long hydrophobic signal sequence and the second occurs at a junction between two basic amino acids and a small polar amino acid. A notable exception is the cleavage at the N-terminus of the dengue NS3 which may take place at the junction between Gln Arg and Ser. Comparative analysis suggests that dengue NS3 and NS5 may be involved in enzymatic activities related to viral replication and/or transcription. Putative nonstructural proteins NS2a, NS2b, NS4a, and NS4b are extremely hydrophobic, suggesting that these proteins are most likely associated with cellular membranes. PMID- 3039727 TI - [Effect of immunization with an inactivated influenza vaccine, NIVGRIP, on the viral population and clinical course of patients with chronic non-bacterial pharyngitis]. AB - Study was conducted on 20 subjects with non bacterial chronic rhinopharyngitis. Forty-one virus strains, mainly adenoviruses, type 3 parainfluenza virus, influenza and type 1 herpes viruses, as well as enteroviruses were isolated from samples collected at regular intervals. After the NIVGRIP (R) vaccination, the number of subjects with viruses present in the pharynx decreased by 75% (from 20 to 5 positive patients) and that of isolates from 41 to 6 (80% positive samples before vaccination against 10% after immunization). PMID- 3039729 TI - The fates of undermethylated mRNA cap structures of vesicular stomatitis virus (New Jersey) during in vitro transcription. AB - A dual label pulse-chase analysis employing S-adenosyl-L-[methyl-3H]methionine and [beta-32P]GTP has been devised to identify precursors to the cap structure 7mG(5')ppp(5')Am present on the 5'-termini of vesicular stomatitis virus mRNAs. Both monomethylated cap structures, 7mG(5')ppp(5')A and G(5')ppp(5')Am, have been detected in vitro in New Jersey serotype reactions containing suboptimal concentrations of S-adenosyl-L-methionine. The simultaneous chasing of both radiolabeled substrates allowed the determination of the transcriptive fate of each pulse-labeled cap structure in the total RNA population. Ten percent of the 7mG(5')ppp(5')A cap structure and 27% of the G(5')ppp(5')Am cap structure generated during the pulse were chased into 7mG(5')ppp(5')Am. These results suggested that while there may have been a preferred order of 5'-cap methylation, the order was not compulsory. The dual label analysis also revealed that only 34% of the pulse-labeled G(5')ppp(5')A cap structure could be chased into methylated cap structures. A nascent RNA chain length-dependent "methylation window" is proposed. PMID- 3039730 TI - A novel method for obtaining poliovirus 14 S pentamers from procapsids and their self-assembly into virus-like shells. AB - Poliovirus procapsids, isolated and partially purified from infected HeLa cells, were found to dissociate into 14 S pentamers following freezing at -20 degrees overnight. Unlike pentameric subunits isolated directly from virus-infected cells or those obtained by alkaline treatment of procapsids, the "freeze-dissociated" subunits self-assembled at room temperature into two kinds of empty capsids, one of which was similar to the shell of poliovirions as shown by its neutral pl, resistance to alkaline dissociation, and the presence of neutralization-related (N) epitopes. The other empty shell resembled that which was previously described as a self-assembled empty capsid (SAEC), as indicated by its acidic pl and absence of N antigenic determinants. At 37 degrees, the assembly reaction was significantly faster but only SAEC were formed. By following the assembly reaction by electron microscopy, intermediate structures were visualized; their relatively low amount at any given time was consistent with the previous hypothesis (J.R. Putnak and B.A. Phillips (1981), J. Virol. 40, 173-183) that initiation is the rate-limiting step in the morphogenesis of empty shells. The relevance of these findings to the morphogenesis of picornaviruses, and polioviruses in particular, is discussed. PMID- 3039731 TI - 72-bp element contains a critical control region for SV40 late expression in Xenopus laevis oocytes. AB - The SV40 late promoter is transcribed at least 10-fold more efficiently than the SV40 early promoter when SV40 DNA is injected into the germinal vesicle of Xenopus laevis oocytes. Late expression in the oocyte is independent of T antigen and does not require DNA replication. To identify DNA sequences required for SV40 late gene expression, 12 mutants spanning nucleotide position (np) 5187 to np 304 were injected into the germinal vesicles of X. laevis oocytes, and RNA was extracted 18 to 24 hr later. S1 nuclease analysis of the 5' ends of SV40 late mRNA revealed that mutations in the origin of replication had no quantitative or qualitative effect on the 5' late start sites. Mutants which deleted the 21-bp repeats did not reduce or alter use of the major RNA initiation site (np 295), but did reduce use of a minor initiation site within the 72-bp repeats. In contrast, deletion of or certain point mutations in the 72-bp repeat decreased initiation from the major late start site. An 85-bp insertion containing a complete set of the 21-bp repeats positioned to the late side of the enhancer elements also decreased initiation from the major late start site. Thus, an element in the 72-bp repeat appears to be the major promoter element for late SV40 transcription in the oocyte. PMID- 3039732 TI - The synthesis of a particle-forming cellular protein is enhanced by Mengo virus infection. AB - We have detected a cellular protein which not only escapes the shutoff of host translation induced by Mengo virus, but is synthesized in increasing amounts during Mengo virus infection. The protein has an apparent molecular weight of 20,000 and is contained in a remarkably stable cytoplasmic particle with a sedimentation coefficient of approximately 16 S in sucrose gradients. In the electron microscope this particle appears as a sphere of approximately 12 nm diameter. The synthesis of the protein is stimulated in mouse L cells and in HeLa cells infected with Mengo virus. Its synthesis cannot be induced, however, by stress (heat) or by infection with reovirus. PMID- 3039733 TI - Molecular analysis of integrated human papillomavirus 16 sequences in the cervical cancer cell line SiHa. AB - Human papillomavirus (HPV) 16 is frequently found integrated into cervical cancer cell genomes and these integrations are thought to play a role in tumorigenesis. To investigate the mechanisms of HPV integration and its effect on transcription and chromosomal sequence organization, we have cloned and analyzed the HPV16 integration from the cervical cancer cell line SiHa. Restriction analyses and Southern blotting indicated that approximately 95% of an HPV16 genome was integrated without gross rearrangement. Sequence analysis of the cellular-viral DNA junctions revealed that integration had occurred within the E2 and E4 ORFs where 251 bp of viral sequence was deleted. One viral terminus occurred within sequences of an Alu repeat and a 4-bp homology was present at the site of recombination. Using unique cellular flanking DNA probes, a 4.8-kb deletion of cellular sequences was detected at the site of viral integration. The chromosomal location of the viral integration and cellular deletion were mapped to chromosome 13 using a rodent X human somatic cell hybrid panel. Northern blot analysis using viral subgenomic and 3' cellular probes revealed transcription from the 3' portion of integrated HPV16 (E6, E7, E1) and flanking cellular sequences. The observation of viral-cell transcripts and chromosomal deletions associated with HPV integration may indicate that such events are part of a multistep mechanism leading to the development of cervical cancer. PMID- 3039734 TI - [Endocrine changes in cancer of the rectum and colon]. PMID- 3039735 TI - [Debatable issues of the histogenesis of malignant fibrous histiocytoma]. PMID- 3039736 TI - [Prognostic importance of the tumor receptor status in patients with ovarian cancer]. AB - Estrogen (ER), progesterone (PR), androgen (AR) and glucocorticoid (GR) receptor levels were assayed in 70 malignant epithelial ovarian tumors from untreated patients and in 23 malignancies from those receiving preoperative chemotherapy. Both PR+ and AR+ frequency and their mean levels in tumor were found to be relatively lower after preoperative chemotherapy. The prognostic significance of receptor status in tumor was identified: the efficacy of combined chemotherapy in PR- and RA- tumor patients was much lower as compared with PR+ and RA+ tumors; a 2-year recurrence-free survival of 12.5% and median duration of remission of 6 months were observed in the former group, while in the latter the disease-free survival rate was as high as 66.7%, and a median duration of remission had not been reached at 24 months of follow-up. PMID- 3039737 TI - [Rare combination of breast cancer and sarcoma (a case)]. PMID- 3039738 TI - [Function of the hypophyseal-ovarian-adrenal system in trophoblastic disease]. AB - Hyperproduction of chorionic gonadotropin was shown to be the first link in the chain of hormonal disturbances involved in trophoblastic disease, which induced changes of pituitary gonadotropic function. Ovarian function is affected by disturbances in trophoblastic and pituitary gonadotropic functions and depends on the extension and severity of the disease. This makes a case for corrective hormonal therapy in chorionepithelioma patients resistant to cytostatic or substitution therapy with sex hormones and in whom ovaries have been removed during combined treatment. PMID- 3039739 TI - [Possible mechanisms of disorders of GTPase activity with substitutions at the 12th and 61st codons of the p21 H-ras protein]. PMID- 3039740 TI - [Effect of Salvia sclarea essential oil on indices of the hematopoietic, immune and enzyme systems]. PMID- 3039742 TI - [Serum copper level in children with malignant neoplasms]. PMID- 3039741 TI - [Clinico-laboratory, immunologic and radionuclide joint indices in psoriatic arthropathy]. AB - A juxtaposition between the clinical-laboratory, immunologic and radionucleotide articular parameters was performed in 50 patients with psoriatic arthropathy, distributed according to the incidence of X-ray manifestations. The late and moderate changes in ESK, leukocytes, fibrinogen, DPA and phosphatasemia do not characterize the severity of the disease. A tendency to hyperuricemia and hypercalcemia is established in the period of arthralgia before the X-ray image for bone-tissue damage. The genetic HLA-B27 predetermination plays a certain role for the more frequent involvement of the spine and sacroiliac joints in the pathological process. The disturbances in the immune system are manifested with increased number of early T-lymphocytes, FcG-receptor lymphocytes, complement (C3b)--receptor lymphocytes and spontaneously blast-transformed cells. The increasing IgG content correlates with the accumulation of macromorphological X ray images and with the index of mineral metabolism from the articular study with 99MTc-pyrophosphate. The changes in the pertechnetium index for articular vascularization are quantitatively insignificant and do not allow the joining of the psoriatic arthropathy to the group of primary synoviarthritis. The accumulation of technetium pyrophosphate in the articular structures is asymmetric, focal and precedes the changes in the X-ray image. PMID- 3039743 TI - [Mechanism of action of benzodiazepines]. PMID- 3039744 TI - [A case of crystallization of urinary calculi on dexon sutures]. PMID- 3039745 TI - [Results of surgical treatment of malignant brain tumors]. AB - Although surgical treatment seldom cures any malignant brain tumours, it remains the basis of the management of these tumours. Studies by others and our own statistical studies demonstrate the absence of any real or significant improvement in survival time for most of the patients. Hence we must await progress in other oncological disciplines to provide a solution. PMID- 3039746 TI - [Local hyperthermia of malignant brain tumors]. AB - In glioblastoma multiforme and high-grade astrocytomas the following procedure has been implemented to try and prevent recurrence: after extirpation of the tumour its bed is lined with metal powder and the patient's head is exposed to a high-frequency magnetic field every 4 weeks. As a result the temperature is raised by induction to +45 degrees C at the boundary between metal and brain tissue. This ought to be sufficient to prevent tumour recurrence. Varied results were achieved using this technique in 103 patients. PMID- 3039747 TI - [A 42-year-old patient with a metastasizing insulinoma]. AB - We report the case of a patient suffering from a rapidly metastasizing insulinoma who after a period with noncharacteristic symptoms suddenly fell into confusional states leading to initial misinterpretation. PMID- 3039748 TI - [Endocrine pancreas tumor as a cause of exocrine pancreas insufficiency: problems of immunologic tumor diagnosis and classification]. AB - The case of a 32-year old man with exocrine pancreatic insufficiency and progressive weight loss is reported. Ultrasound examination of the abdomen revealed a huge tumor of the pancreatic head. Smears obtained by a percutaneous fine-needle biopsy showed characteristics of a neuroendocrine tumor. Serum hormone levels and immunocytochemistry presented no evidence for a hormonal activity of the tumor. The exocrine pancreatic insufficiency was explained by an obstruction of the pancreatic duct system. In the present case report the problems of diagnostic approaches to endocrine pancreatic tumors are discussed. PMID- 3039749 TI - [Transition to cutaneous squamous cell carcinoma in 2 patients with bowenoid papulomatosis]. AB - We report on two female patients (30 and 33-year-old) suffering from bowenoid papulosis (BP) which showed HPV-16-DNA (E.-M. de Villiers, DKFZ, Heidelberg). Both patients developed cutaneous invasive squamous cell carcinoma in the perineal and vulvar region, resp. In the first patient, the microinvasive part of the tumor was excised. Subsequent therapy with interferon-gamma resulted in complete clinical and histological remission of the BP within 4 months; 6 months after discontinuation of the therapy with interferon, however, we observed a relapse of BP. In the second case, which showed more prominent BP alterations and deeper tumor invasion, interferon-gamma did not bring on any improvement within 3 weeks of therapy; thus radical operation was performed. To our knowledge, this is the first report on the transition of BP of the outer ano-genital region into invasive squamous cell carcinoma. PMID- 3039750 TI - [The interface between hydroxyapatite ceramic and newly formed bone in scanning electron microscopy]. AB - Bone samples, which have been embedded in methylmethacrylate in an undecalcified state can be evaluated by a scanning electron microscopy after removal of the methylmethacrylate. Hydroxy apatite (HA) has been implanted in 6 mm bore holes of the rabbits' femoral condyle. The animals were sacrificed after 2, 4, 6, 8 weeks and 8 months. The interface between HA and the newly built bone was observed. A direct contact has been found regularly through high magnification (up to 16,000 times). The three dimensional embedding of the implanted HA-particles into the newly built bone can be shown. Collagen filaments are spreading out from bone to HA-particles. We think that there is not only a mechanical but also a chemical connection between both. PMID- 3039751 TI - Effects of cytoskeletal disrupting agents on mouse mammary tumor virus replication. AB - Alterations in mouse mammary tumor virus (MMTV) production and composition were induced by exposure of mammary tumor cells to cytodisruptive agents. Treatment with 2.1 microM cytochalasin D (CD) for 24 h reduced MMTV yield by 80% and electron microscopic examination of these cells did not reveal budding virions. Immune precipitation and quantitative immunofluorescence studies demonstrated that CD had no significant effect on MMTV polypeptide synthesis or surface expression suggesting that CD inhibited late steps in MMTV maturation. Decreases in MMTV production were also observed as a result of 24 h exposure of the cells to 2.1 microM cytochalasin B (CB). However, an initial 70% increase in the levels of extracellular virions within the first 18 h of treatment preceded diminution of virus production. In addition, CB was unable to abrogate maturation and release of MMTV particles as revealed by electron microscopic evaluation of thin sections of treated cells. Colcemid at 0.28 microM had no effect on virus production during the first 24 h of exposure although MMTV yield was reduced by 60-70% after 36 h of treatment. Polypeptide profiles of MMTV purified from cell cultures treated with any of the three cytodisruptive agents were altered and included 5-7 polypeptides not typically present in MMTV from untreated cells. These cytodisruptive agents did not significantly affect viability and protein metabolism of MJY-alpha cells; the data suggest that alterations in MMTV replication were due to disruption of the cellular cytoskeleton. PMID- 3039752 TI - Induction of antibody against in vitro translation products encoded by varicella zoster virus glycoprotein genes. AB - Antibodies were raised in rabbit against the in vitro translation products encoded by the varicella-zoster virus (VZV) glycoprotein genes gpI and gpIV. The antisera neutralized VZV infectivity and specifically identified two late VZV glycoproteins, gpI and gpIV, in VZV-infected cells and in the envelope of VZ virions. Pulse-chase experiments revealed a 55K precursor protein to gpIV (60K) and a 82K precursor protein to gpI (95K). Immunoprecipitation of 32P-labeled VZV infected cells showed that the precursor-products of gpI are phosphorylated. These results demonstrate that translation products synthesized in vitro can be used to produce antibodies that recognize native viral proteins and therefore facilitate the identification and analysis of viral gene products in the infected cells. PMID- 3039753 TI - [Spontaneous nephroblastomas in rats]. PMID- 3039755 TI - [Prosthesis-epithesis treatment after complete loss of the maxilla with an extensive facial defect]. PMID- 3039754 TI - [The rejection reaction in subcutaneous connective tissue in the rat after application of silicon dioxide]. PMID- 3039756 TI - [Cystosarcoma phyllodes of the lacteal glands]. AB - Thirty-three cases of phyllode cystosarcoma were diagnosed in the course of 21 years at the Tumour Department of the Berlin School of Medicine (Charite). Ten of them were found to be in sarcomatous degeneration. Metastasation was positively established in four of these ten cases, and locally delimited recurrences were recorded from two. Metastasation in cases of phyllode cystosarcoma has been found to occur rarely through regional lymph nodes. Therefore, routine removal of those lymph nodes is not considered necessary. PMID- 3039757 TI - [Case report of a malignant chemodectoma of the lower anterior mediastinum]. PMID- 3039758 TI - [Vulvar cancer originating from pointed condylomas--case report]. AB - A vulvar cancer is demonstrated. Its origin from condylomata acuminata is discussed. The ultrastructural evidence of papillomavirusparticles within the nucleus of koilocytes are hints at a possible human papillomavirus etiology of vulvar carcinoma. Because of the malign transformation of condylomata acuminata described we recommend a consequent treatment and histologic examination of condylomata acuminata. PMID- 3039759 TI - [Course of pregnancy and labor following cytostatic treatment of trophoblastic tumors]. AB - In our department 24 patients underwent treatment of invasive trophoblastic tumours from 1975 to 1984. In 13 cases we were dealing with invasive bladder moles and in 11 cases with chorionic carcinoma. Nine women became pregnant, among them one case of chorionic carcinoma. Eight women were delivered of a viable child on average 30 months after the end of the treatment, whereas one patient wanted termination of pregnancy. Six women had undergone a methotrexate monotherapy before, two women a sequential treatment with methotrexate + actinomycine D. The undisturbed pregnancies and births of eight healthy babies point to an unimpaired fertility after chemotherapy of trophoblastic tumours. PMID- 3039760 TI - [Hypertrophic polyneuropathy in cats]. PMID- 3039761 TI - [Present occurrence of parainfluenza virus 2 in hybrid laying chickens]. PMID- 3039762 TI - A direct neutralizing peroxidase-linked antibody assay for detection and titration of antibodies to bovine viral diarrhoea virus. PMID- 3039763 TI - [Cadmium distribution in tissues and Na,K-ATPase activity of the skin of the frog Rana temporaria in different routes of cadmium uptake by the body]. AB - Accumulation, distribution of Cd in tissues and its toxic effect depend of the mode of uptake of Cd by the organism. Subcutaneous injections of Cd (0.12-0.24 mg/100 g per day) for 10 days resulted in a significant accumulation of Cd in the liver and kidney. No death cases were observed. Addition of CdCl2 to the aquatic environment (more than 0.002%) caused acute toxic effect on frogs. Within 10 days, significant amounts of Cd were found in the skin, especially in its outside layers, small amounts were found in the kidney and liver. High external concentrations of CdCl2 (0.005%) inhibited the activity of Na,K-ATPase in the skin epithelium. It is suggested that adaptive detoxication of cadmium in the liver and kidney operating in mammals, is ineffective in amphibians provided cadmium uptake occurs via the skin. PMID- 3039764 TI - [Functional macrophage activity in infection with virulent and avirulent strains of the dysentery microbe]. AB - The effect of S. flexneri virulent and avirulent (vaccine) strains 2a on the cytoplasmic membrane of mouse macrophages has been studied by evaluating the action of these bacteria on the activity of 5-nucleotidase. The dynamics of the activity of 5-nucleotidase after the introduction of both virulent and avirulent strains has a phasic character with alternating rises and falls in the activity of this enzyme in comparison with the control. S. flexneri vaccine strain produces mainly a stimulating effect on the functional activity of peritoneal macrophages in mice, which is confirmed by a decrease in the activity of 5 nucleotidase; on the contrary, S. flexneri virulent strain- has mainly an inhibiting effect on the functional activity of peritoneal macrophages, which is confirmed by an increase in the activity of 5-nucleotidase in these cells. The comparative study of changes in the activity of 5-nucleotidase, following the introduction of S. flexneri, in mice, previously immunized with smallpox vaccine, and in intact mice has shown that the use of animals immunized with smallpox vaccine in the study of metabolic characteristics may lead to distortions in the results of the experiment. PMID- 3039765 TI - Continent ileostomy. A follow-up study of 60 patients. AB - Sixty patients underwent proctocolectomy and received a continent ileostomy because of ulcerative colitis (50 cases) familial polyposis (7) or Crohn's disease (3), with no deaths at surgery or during follow-up (mean 4.5 years, range 3 months-10 years). Early complications were few and insignificant. Late complications (nipple-valve sliding and pouch ileitis) were more common, and 15 patients with valve sliding causing leakage had to undergo altogether 22 reoperations. Three reservoirs had to be removed because of refractory pouch ileitis. Modifications in nipple-valve construction in the last 40 cases diminished the problem of sliding. More than 90% of the patients reported improved quality of life after conversion of conventional to continent ileostomy. Continent ileostomy remains an excellent alternative to ileoanal anastomosis with proximal reservoir for patients who cannot accept conventional ileostomy or when an ileoanal anastomosis with reservoir functions unsatisfactorily or is otherwise unsuitable. PMID- 3039766 TI - Effects of fludrocortisone withdrawal on plasma angiotensin II, ACTH, vasopressin, and potassium in patients with Addison's disease. AB - We attempted to answer to the question whether excessive rises in endogenous plasma angiotensin II (AII) stimulate ACTH secretion by measuring PRA, AII, AVP, ACTH, and cortisol in 8 patients with Addison's disease before and after withdrawal of fludrocortisone substitution. Blood was drawn at 14.30 h, exactly 6 1/2 h after the morning dose of hydrocortisone had been taken. PRA and AII were initially higher than normal in 4 patients. After withdrawal of fludrocortisone for 1 or 2 weeks, PRA and AII rose markedly in 4 patients (up to 260 ng/l) without concomitant changes in plasma ACTH levels (r = -0.081, AII vs ACTH). Changes in plasma cortisol could not have obscured a stimulatory effect of AII on ACTH by variable feedback inhibition of ACTH release. The increase in plasma AII levels in the 4 patients was larger than that observed in a previous study in normal subjects after rigorous dietary sodium restriction. In all patients, hyperkalaemia developed after fludrocortisone withdrawal, independent of changes in PRA and AII. Rises in PRA, AII, and plasma potassium were partially reversed by increased sodium intake and further suppressed by resumption of fludrocortisone therapy. Plasma AVP remained in the normal range after fludrocortisone withdrawal, but was slightly elevated after increasing salt intake without fludrocortisone administration. CONCLUSIONS: Rises of endogenous plasma AII to levels tenfold higher than normal do not stimulate ACTH release. AII is probably not a physiological modulator of ACTH secretion. Mineralocorticoid substitution in Addison's disease should be monitored by plasma potassium measurement. Hyperkalaemia may coexist with normal PRA.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3039767 TI - Vitamin D3 metabolism in a pig strain with pseudo vitamin D-deficiency rickets, type I. AB - Vitamin D metabolism was studied in the 'Hannover Pig', a strain which suffers from pseudo vitamin D-deficiency rickets, type I. Animals of this strain are known to be devoid of renal 25-hydroxyvitamin D3-1 alpha-hydroxylase and -24 hydroxylase activities. Pigs with florid rickets and hypocalcaemia were treated with single im injections of 0.25 to 1.25 mg of vitamin D3, doses that have been shown in previous studies to be effective in producing transient healing of rachitic symptoms. The levels of vitamin D3 and its most relevant physiological metabolites in plasma were estimated at intervals before and after this vitamin D3 treatment. Vitamin D3 rose from 14.8 +/- 8.1 to 364 +/- 190 nmol/l (mean +/- SD) 2 to 3 days post injectionem, 25-hydroxyvitamin D3 from 131.0 +/- 46.2 to 1068 +/- 160 nmol/l within 7 days post injectionem. The 1 alpha, 25 dihydroxyvitamin D3 concentration in plasma was elevated from 73.9 +/- 25.0 to 281 +/- 168 pmol/l 2 to 3 days post injectionem and declined continually from that time. 24R,25-dihydroxyvitamin D3 and 25S,26-dihydroxyvitamin D3 levels after treatment showed different responses in different animals being either elevated or unchanged. Clinical healing of the pigs with these doses of vitamin D3 was attributed to the transient rise of 1 alpha,25-dihydroxyvitamin D3 in plasma. It was assumed that 1 alpha,25-dihydroxyvitamin D3 synthesis takes place under these circumstances in extrarenal tissues. PMID- 3039768 TI - Is thyrotropin-releasing hormone receptor involved in thyrotrope adaptation to starvation? AB - The aim of the present study was to delineate the involvement of TRH receptors in the thyrotrope adaptation to starvation (i.e. plasma TSH and thyroid hormone decrease, increased sensitivity to T3) by measuring [3H]TRH binding in euthyroid, hypothyroid and T3-substituted rats (175 ng/100 g body weight). Our results show that in euthyroid rats, starvation does not significantly modify either the affinity or the number of pituitary binding sites. In hypothyroid and T3 substituted rats, starvation does not alter the negative control exerted by T3 on the number of TRH binding sites. Our data indicate that the adaptation of thyrotrope to starvation does not primarily result from alterations of TRH binding sites. PMID- 3039769 TI - Effect of vitamin D2 and vitamin D3 on the serum concentrations of 1,25(OH)2D2, and 1,25(OH)2D3 in normal subjects. AB - Serum concentrations of vitamin D2 and vitamin D3 metabolites were measured in 19 normal subjects before and during treatment with either vitamin D2 or vitamin D3, 4000 IU per day for 8 weeks. Vitamin D2 treatment increased the serum concentration of 1,25(OH)2D2, but a corresponding decrease in 1,25(OH)2D3 resulted in an unchanged serum concentration of total 1,25(OH)2D. During treatment with vitamin D3, the serum concentration of 1,25(OH)2D metabolites was unchanged. We conclude that the production of 1,25(OH)2D is tightly regulated and that 1 alpha-hydroxylase does not discriminate between D2 and D3 metabolites in normal subjects. PMID- 3039770 TI - Detection of late-onset adrenal hyperplasia in girls with peripubertal virilization. AB - We investigated the value of serum levels of adrenal steroids (dehydroepiandrosterone sulphate, testosterone, 17-hydroxyprogesterone, cortisol) in the identification in peripubertal females with late-onset congenital adrenal hyperplasia owing to 21-hydroxylase deficiency. Among 68 females (age 3-18 years) with virilization in childhood, peripubertally or postpubertally, we selected 21 girls for an ACTH test by measurement of basal blood-spot or serum 17 hydroxyprogesterone (17-OHP) levels. Eight of 21 patients had supranormal post ACTH serum 17-OHP concentration (57-153 nmol/l) with low normal cortisol concentration. All of them had supranormal basal and post-ACTH 17-OHP to cortisol ratios. These data show a relatively high incidence (about 12%) of mild 21 hydroxylase deficiency among prepubertal and adolescent girls with virilization. It is concluded that the first step in the investigation of peripubertally virilized girls should be the determination of serum 17-OHP and cortisol. Patients with basal morning 17-OHP concentration and 17-OHP to cortisol ratio above reference range should be given an ACTH test. PMID- 3039771 TI - Restriction enzyme analysis of HLA class II DR beta genes in patients with Graves' disease. PMID- 3039772 TI - Polymorphism of the immunoglobulin heavy chain T cell receptor beta-chain genes in Graves' disease. PMID- 3039773 TI - Immunoreactivity of PTH-binding in intact bovine kidney tissue and cultured cortical kidney cells indicative for specific receptors. AB - Localization of PTH-binding sites has been examined in intact kidney sections and cultured cells derived from bovine kidney cortex. Tissue sections were incubated with 10(-7) M bovine PTH (1-84) for 2 h, cells for 15 min, at 37 degrees C. Visualization of PTH-binding was achieved by immunocytochemistry using a carboxy terminal specific anti-PTH antiserum (S 478). For control, cell culture incubations were performed applying competitively 10(-7) bovine PTH (1-84) and a 10-fold excess of synthetic 1-34 PTH fragment, not antigenic for S 478. This resulted in a lack of staining. PTH-binding was found in all cells of the proximal and the distal tubule, and with less intensity in the thick ascending limb of Henle's loop. In collecting ducts a PTH specific staining was also present, which was confined to single cells localized between others without PTH binding sites. No staining was seen in glomerula, the thin limb of Henle's loop, in blood vessels, and in connective tissue. The data suggest that large parts of the nephron contain PTH-binding sites, although in different amounts. This is in agreement with the numerous actions of PTH in the kidney. In the collecting segment a distinct cell-to-cell difference was disclosed indicative for different functional states or cellular heterogeneity. PMID- 3039774 TI - Control of thyroid cell proliferation: the example of the dog thyrocyte. PMID- 3039775 TI - Role of the adenylate cyclase-cAMP system on TSH-stimulated thyroid cell growth. AB - TSH is a trophic factor for cultured rat thyroid cells (FRTL-5). In the present study we have investigated the mechanism by which TSH promotes cell growth and evaluated the possible role of the adenylate cyclase (AC)-cAMP system in this process. The mitogenic activity of several agents was evaluated by measuring their effect on cell number or 3H-thymidine incorporation into DNA. Forskolin and cholera toxin, two potent and specific activators of the AC, induced a dose dependent increase of 3H-thymidine incorporation. The maximal stimulation, observed at concentrations of 10 microM and 10 ng/ml, respectively, was beta 80% of that obtained with optimal concentrations of TSH. A similar effect was obtained with a Graves' IgG preparation (0.2 mg/ml) able to stimulate the thyroid AC or with 3-isobutyl-1-methyl-xanthine (IBMX, 0.5 mM), a phosphodiesterase inhibitor. 8-bromo cAMP (0.5 mM), a cAMP analog, also stimulated 3H-thymidine incorporation, and its potency was approximately 60% of that of TSH. Similar results were obtained when the mitogenic activity of these compounds was evaluated by cell number. Norepinephrine (NE, 10 microM), although devoid of AC stimulatory activity in these cells, also stimulated 3H-thymidine incorporation, but its potency was only 20-30% of that of TSH. Indomethacin (100 microM), an inhibitor of phospholipid and arachidonic acid metabolism, was able to inhibit the stimulatory effect of NE (84%), and to a lesser extent of TSH (63%) and cholera toxin, had minor effect on forskolin (24%), IBMX (16%) and Graves' IgG (8%), and no effect on 8-bromo cAMP.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3039776 TI - Inhibition of cAMP formation by EGF in thyroid follicles is mediated by intracellular Ca++. AB - The mechanism of cAMP-inhibition by EGF was studied in isolated porcine thyroid follicles. EGF inhibited TSH-induced cAMP-formation maximally by 40%, this effect remained up to 1 h of pre-incubation. The calcium-ionophore A 23 187 also inhibited cAMP-formation, but its effect was relieved after 1 h. The phorbolester TPA had a biphasic influence on cAMP-formation, with a transient increase (5 min) before a sustained inhibition (60 min); the inhibitory effect was mimicked by the diacylglycerol 1-oleoyl-2-acetyl-glycerol. Exogenous arachidonic acid had only a small and transient inhibitory effect on cAMP-formation. We conclude, that EGF inhibits cAMP-formation by a raise of intracellular Ca++, as well as by the direct activation of proteinkinase C, indicating, that a phosphorylated product could be a mediator for the inhibition of adenylate cyclase. PMID- 3039777 TI - Sodium/proton exchange in FRTL-5 thyroid cells: role in the maintenance of intracellular pH and TSH-stimulated cell growth. PMID- 3039778 TI - Triple alpha genes in association with sickle cell and beta-thalassaemia gene in the Saudi population. AB - This paper describes the case of a 6-year-old Saudi male who had sickle cell heterozygosity, beta +-thalassaemia and possessed three alpha-genes of the haplotype alpha alpha alpha anti-3.7/as diagnosed by restriction endonuclease studies using Hpa I, Bam HI, Bgl II, Hind III and Xba I. Since the iron level was found to be normal, it is proposed that the coexistence of beta-thalassaemia with triple alpha-genes in Hb S heterozygotes may be the cause of the anemia. A possible mechanism for severe anaemia is presented. PMID- 3039779 TI - Histomorphometric study of the superficial peroneal nerve in leprosy. AB - A histomorphometric analysis of the superficial peroneal nerve was made in 15 cases of leprosy. Thirteen patients presented clinical signs of leprous neuropathy, while the other two showed only cutaneous signs of leprosy. The presence of M. leprae in all the nerves sampled, and the appearance of the histologic lesions, made it possible to confirm the diagnosis of leprosy and to specify the type of leprosy in each case, even in the absence of clinical signs of leprous neuropathy. Correlation of the histomorphometric results with the duration of development of the disease and with the time elapsing before treatment showed, in the beginning stage, a considerable reduction in myelinated and unmyelinated never fibres and a proliferation of Schwann cells, as well as segmental demyelination and axonal degeneration of the teased fibres. When treated early, the evolution of the lepromatous type (LL) appears favourable, with apparent regeneration of the nerve fibres. When treatment is not instituted early, gradual loss of nerve fibres, axonal degeneration of all the teased fibres, proliferation of the Schwann cell processes devoid of axons, and increase in endoneural connective tissue lead to a severe degeneration of the nerve. This unfavorable development appears to progress faster in the absence of treatment or when treatment is irregular. In the borderline lepromatous (BL) and borderline tuberculoid (BT) types, nerve degeneration appears to be more rapid than in the type LL. PMID- 3039780 TI - Localized outbreak of enteroviral meningitis in adults. AB - During a thorough surveillance of viral infections of the central nervous system an outbreak of aseptic meningitis was discovered in the western part of Finland in late 1985. The 21 diseased young adults were carefully studied by different virological methods. A presumed viral etiology, in all cases of enteroviral origin, was found in 16 of 20 (80%) with adequate specimens. Four different enteroviruses were associated with this episode; in 9 cases the presumed etiological agent was echovirus 5, while coxsackie B5, echo 25 and echo 17 viruses appeared to be responsible for 4, 2 and 1 case, respectively. Sensitivity of different diagnostic methods as regards detection of the echovirus 5 infections was in order: increase of type-specific neutralizing serum antibodies, isolation of virus from faeces, isolation from throat and group diagnosis by demonstrating an increase in complement-fixing antibodies to coxsackie B5 virus antigen. PMID- 3039781 TI - Leukotrienes B4 and C4 in MS. AB - Release of leukotriene B4 (LTB4) and leukotriene C4 (LTC4) from neutrophils and platelet-neutrophil suspensions in response to ionophore A23187 was measured in 12 multiple sclerosis (MS) patients and 8 healthy volunteers. LTC4 release from neutrophils, as well as from platelet-neutrophil suspensions, was significantly decreased in MS patients compared with the controls. There was no significant difference in the release of LTB4 between MS patients and controls. The findings suggest that permanent stimulation of platelets and neutrophils e.g., by encephalitogenic peptide leads to continuous LTC4 release with subsequent depletion of intracellular substrates serving as precursors for the formation of 5-lipoxygenase products. Since the target of microvascular actions of LTC4 are postcapillary venules, the release of this sulfidopeptide leukotriene might play a pathogenetic role in the formation of MS lesions. PMID- 3039782 TI - Variability of morphological features in early infantile polyneuropathy with defective myelination. AB - Four cases of early infantile polyneuropathy with defective myelination are reported. The peripheral nerve was studied by light and electron microscopy; different morphological characteristics have been noticed in these patients. Case 1 presented aspects of defective myelination with atypical "onion bulb" formation composed of multiple layers of basement membrane. In case 2, defective myelination and atypical "onion bulb" formation were associated with aberrant hypermyelination. Cases 3 and 4 were brothers, who presented axonal damage and atypical "onion bulb" formation. PMID- 3039783 TI - Proliferative potential of vascular components in human glioblastoma multiforme. AB - Sixteen patients with glioblastoma multiforme received a 1-h intravenous infusion of 5-bromo-deoxyuridine (BrdU), 150-200 mg/m2 at the start of surgery, to label S phase cells in tumor tissue. Labeled cells of vascular components and of tumor parenchyma were detected in excised tumor specimens by indirect immunoperoxidase staining using anti-BrdU monoclonal antibodies followed by periodic acid-Schiff staining. The BrdU labeling index (LI, defined as the percentage of labeled cells in relation to the total number of cells scored) was calculated separately for vascular components and tumor parenchyma in each specimen. The BrdU LI of vascular components of glioblastoma multiforme was remarkably higher than that of normal brain (1.1%-8.7% vs. less than 0.05%). The mean BrdU LIs of vascular components and tumor cells in eight primary glioblastomas were 4.5 +/- 0.8% (mean +/- SE) and 9.9 +/- 1.1%, respectively, while the corresponding BrdU LIs in eight recurrent tumors were 2.7 +/- 0.5% and 9.3 +/- 0.7%. The differences in the BrdU LIs of primary and recurrent tumors were not statistically significant, but the BrdU LI of vascular components was consistently much lower than that of tumor cells. BrdU labeling of vascular components was inconsistent and occurred mostly in glomerular-shaped vessels, but only about 20% of them contained labeled cells. These results suggest that unusual vascular proliferation, such as the formation of glomerular-shaped vessels and endothelial or adventitial proliferation, in glioblastoma multiforme may have been programmed to slow down or even to cease at a certain stage, and is not likely to be the result of neoplastic transformation. PMID- 3039784 TI - Immunohistochemical staining of cells in the brain of a patient with acquired immune deficiency syndrome (AIDS) with a monoclonal antibody to visna virus. AB - Several monoclonal antibodies to different epitopes of the two major core proteins of visna virus, p25 and p15, were tested with the Avidin-Biotin immunostaining method on formaldehyde-fixed and paraffin-embedded sections of brains from patients with acquired immune deficiency syndrome (AIDS) who had shown neurological symptoms at death. In one of five AIDS cases a few cells, mainly inflammatory cells, showed a positive staining with a monoclonal antibody to the p25 core protein of visna. PMID- 3039785 TI - Prevalence of condylomatous atypia and human papilloma virus antigen in cervical biopsies in 1972 and 1983. AB - To compare the frequency of pure condylomas and condylomatous atypia in cervical intra-epithelial neoplasia in 1972 and 1983, two equally sized consecutive series of specimens from the uterine cervix were reviewed. The 1972 material contained only 5 cases of pure condyloma, all of these being flat lesions, while the 1983 material contained 33 pure condylomas, 23 flat and 10 papillomatous cases. The prevalence of cervical intra-epithelial neoplasia with condylomatous atypia was significantly greater in the 1983 material. In both years, condylomatous atypia was more frequent in intra-epithelial neoplasia of lower degrees, and patients with intra-epithelial neoplasia with condylomatous atypia were younger than patients with neoplasia only. The proportion of patients with cervical intra epithelial neoplasia with condylomatous atypia showing positive reaction for HPV Ag was also larger in the 1983 material. The results are discussed, especially regarding the possible role of HPV infection in cervical carcinogenesis and the increasing rate of invasive carcinoma of the cervix, observed during recent years. PMID- 3039786 TI - Adenosarcoma uteri. AB - Adenosarcoma of the uterus is a rare tumor of low-grade malignancy. It belongs to the family of mixed mesodermal (Mullerian) tumors. It is a biphasic tumor with benign epithelium and malignant stromal components. A case in a 76-year-old woman is presented. PMID- 3039787 TI - Endocrine responses to head and neck surgery in men. AB - It has been well established that surgical stress leads to profound changes in endocrine function and metabolism. However, the endocrine response varies depending upon the type and the extent of surgery. As no data were available about the endocrine changes during and following major head and neck surgery, this study was performed. Plasma levels of adrenocorticotropin (ACTH), cortisol, thyroid-stimulating hormone (TSH), thyroxin (T4), triiodothyronine (T3), growth hormone (GH), prolactin (PRL), gonadotropins (LH and FSH), oestradiol and testosterone were determined in 17 patients one day before, immediately after, as well as 2 and 4 days after head and neck surgery. An increase in ACTH, cortisol, PRL and GH, and a decrease in plasma oestradiol and testosterone values occurred immediately after surgery. There was a slow fall in cortisol levels after surgery, but they remained elevated even on the fourth postoperative day, whereas GH values returned on the fourth day to the initial level. There were no changes in gonadotropins, TSH and T3, but T4 values were found to be increased on the second and fourth postoperative day. The prolonged cortisol stimulation which was not described by other researchers after other kinds of surgery might be caused by vagal stimulation during and/or after head and neck surgery. Increased needs after a major head and neck surgery could explain the increment of T4 values. PMID- 3039788 TI - [Selective preoperative embolization in the surgery of vascular neoplasms of the nasopharyngeal region]. PMID- 3039789 TI - Fibronectin in human hepatocellular carcinoma (HCC) and HCC cell lines. AB - The localization of fibronectin (FN) in human hepatocellular carcinoma (HCC) was studied in thirty-six HCC tumors (19 autopsy and 17 surgical specimens), two xenografted tumors of HCC to BALB/c mice and three HCC cell lines. The synthesis of FN was also examined in three HCC cell lines. FN was demonstrated on the endothelial surface of the blood spaces of cancerous tissue. Cytoplasmic and intercellular localization of FN was also observed. But there was no correlation between the localization pattern of FN and metastasis. In the two xenografted HCC tumors, FN was found in association with the blood vessels of the tumor tissue and between the HCC tumor cells. In all 3 HCC cell lines, FN was localized on the surface and in the cytoplasm of some HCC cells. FN was detected in the serum-free culture media of three HCC cell lines by immunoelectroblotting. The electrophoretic pattern of FN synthesized by these cell lines was different from that of plasma-FN and resembled that of cellular-FN synthesized by normal liver fibroblasts. PMID- 3039790 TI - "HPV score", a scoring system for histological diagnosis of human papillomavirus infection in dysplasia of the uterine cervix. AB - We devised a scoring system, "HPV score", for histological diagnosis of human papillomavirus (HPV) infection in dysplasia of the uterine cervix. Four hundred and sixty cases of cervical dysplasia were screened for the presence of HPV infection using this system, and 116 cases (25%) were judged to be HPV infection. The results showed good correlation with those obtained using the immunoperoxidase (IMPO) method, but 42 of the 116 cases (36%) were negative for HPV antigen by the IMPO method. Severe stromal inflammation was noted in about half of such cases. Among the four histological types of cervical condyloma, the flat type was most common among the cases with HPV infection judged according to HPV score. It was concluded that this scoring system is simple and accurate for the detection of HPV infection in cervical lesions. PMID- 3039791 TI - Malignant lymphoepithelial lesion. AB - A case of undifferentiated carcinoma arising from benign lymphoepithelial lesion (BLEL) of the parotid gland was studied by light and electron microscopy. Histopathologically, the carcinoma was composed of pleomorphic anaplastic cells showing an undifferentiated type among abundant lymphoid tissue forming germinal center. Among the prominent lymphoid tissue, epithelial hyperplasia, dysplasia, and squamous metaplasia of the duct epithelium were found. Dysplastic epithelium revealed a transition with carcinomatous component in some areas. On the electron microscopic observation, the tumor cells were poorly differentiated, possessing desmosomes and intracytoplasmic filaments. The patient is alive and well 2 months after resection of the tumor, but has a high titer of serum Epstein-Barr virus capsid antigen in IgG. Eighty five cases of the malignant lymphoepithelial lesion (MLEL) including the present case are summarized. PMID- 3039792 TI - Alimentary tract lesions in cytomegalovirus infection. AB - Ten autopsy and 2 biopsy cases of cytomegalovirus (CMV) infection of the alimentary tract were studied. CMV infection was microscopically determined by the presence of cytomegalic inclusion as well as by the immunofluorescent method. Clinical manifestations such as abdominal pain, diarrhea, hematemesis, bloody stool, perforation, and/or abdominal distension with paralytic ileus were observed in 8 autopsy cases and 1 biopsy case. Disappearance of cytomegalic cells was confirmed by the follow-up study in the biopsy cases. Macroscopically, mucosal hemorrhage or ulceration was found in the gastrointestinal tract from the esophagus to the colon. Ulceration showed a characteristic well-defined punched out appearance. The esophagus was the most frequently involved organ. However, no cytomegalic cells were found in the squamous epithelium. In the stomach, regenerated epithelial cells were frequently involved in the deeper part of glands. Numerous endothelial cells transformed into cytomegalic cells in the mucosa surrounding the ulcer in the esophagus, stomach, and intestine. Ischemia caused by cytomegalic changes of vascular endothelial cells is thought to play an important role in the pathogenesis of the ulcer of the gastrointestinal tract. PMID- 3039793 TI - Ultrastructural study of hemangiomas. 5. Glomus tumor (Masson). AB - The fine structure of four cases of glomus tumor was described. All of the cases showed a solid type, and the glomus cells were shown to be typical or modified smooth muscle cells. Our conclusion is that the glomus cells are not derived from pericytes, but from smooth muscle cells of vascular part of the glomus, and are thought to be a specialized smooth muscle cell tumor. PMID- 3039794 TI - Adenovirus hepatitis in a patient with severe combined immunodeficiency. AB - An autopsy case of adenovirus hepatitis with severe combined immunodeficiency (SCID) is presented. A 7-month-old female was admitted to the Kitasato University Hospital with chief complaint of persistent fever. A diagnosis of severe combined immunodeficiency (SCID) was made because of defective function in both T and B lymphocytes. Respiratory disturbance and severe hepatic dysfunction were manifested after admission. She died of respiratory failure on the 40th hospital day. Autopsy findings revealed many focal necroses scattered irregularly throughout the liver. Degenerating hepatocytes around the focal necrosis contained nuclear inclusion bodies, and crystalline arrays of adenovirus virions were recognized in these cells by electron microscopy. Adenovirus antigens were brilliantly detected in the nuclear inclusion bodies by indirect immunofluorescence. PMID- 3039795 TI - Pleomorphic adenoma with a predominantly myoepithelial proliferation of the vagina. AB - A case of pleomorphic adenoma of the vagina in a 44-year-old woman was described. The tumor was a submucosal mass measuring 3.5 X 2 X 2 cm and located in the left lateral wall 1.5 cm inside the introitus. Histologically, it was predominantly composed of sheets and strands of spindle-shaped or polygonal cells focally dispersed in the myxoid tissue to form pseudomicrocysts. Ultrastructure demonstrated the basal lamina, desmosomes, variable amounts of filaments of both intermediate and microfilament, and occasionally microvilli in these tumor cells indicating that dominant cells were myoepithelial in nature. With immunohistochemical studies, keratin and cytokeratin were positive, whereas, S 100 protein, glial fibrillary acidic protein, vimentin, secretory component, and lysozyme were negative. The previously published cases were reviewed and compared with the present case. PMID- 3039796 TI - [Effects of xinchuanling (XC-1) and its derivative XC-2 on adrenoceptors and arterial muscle]. PMID- 3039797 TI - Effect of modification of membrane saccharides on hormonal imprinting in Tetrahymena. AB - Concanavalin-A (Con-A) competes with insulin for the insulin binding sites of the receptor, an event resulting in overlapping bonds. If the saccharide components of the receptor are subjected to periodate treatment the binding of insulin decreases and so does the imprinting evoked by it. Simultaneously, the binding of Con-A increase immediately after the treatment and 24 hours following the insulin imprinting. This phenomenon indicates that though the two ligands (i.e. insulin and Con-A) overlap on the intact receptor, alterations in the saccharide component influence their binding though not in the same manner and not in the same direction. Periodate treatment, similarly as observed in mammalian lymphocyte cultures, enhanced the division of Tetrahymenas and the peak of Con-A binding occurred parallel to the peak of division after periodate treatment of equal duration. Periodate treatment disturbed not only the binding of Con-A but that of other lectins as well. It has been concluded that hormonal imprinting requires a physiological condition of the membrane and any disturbance of the membrane will lead to a decrease in the efficacy of imprinting. PMID- 3039798 TI - Effect of single neonatal melatonin treatment on in vitro thyroxin secretion and TSH or melatonin-modified cAMP level of one-month-old rats. AB - At the age of one month, incubation with melatonin of the thyroid glands of rats having received a single melatonin treatment at the age of three days resulted in increased thyroxine production. TSH was unable to enhance the thyroxine production of animals treated with melatonin neonatally, while its considerable increase could be observed in the case of control animals. Simultaneous TSH and melatonin treatment applied in vitro at the age of one month resulted in an approximately twofold increase of thyroid T4 production in rats having received neonatal melatonin treatment. In vitro alteration of the cyclic AMP level of the thyroid glands of intact and neonatally melatonin treated rats ran practically parallel, except that in the melatonin treated animals the cAMP level was higher after TSH administration. At the same time the cAMP level decreased in the thyroid gland of animals treated with TSH + melatonin. There was no exact correlation between the alterations of cAMP and T4 levels in the given experimental system. PMID- 3039799 TI - Effect of the cAMP level on hormonal imprinting in Tetrahymena. AB - Augmentation of the cAMP level has no positive effect on hormonal imprinting in Tetrahymena. Artificial elevation of the cAMP level may inhibit the development of imprinting or may result in abnormal imprinting. The role of Ca2+ is of great importance in mediation of the imprinting mechanism. Generally, this role is not influenced by an elevated cAMP level but, exceptionally, the latter may effect the mechanism of imprinting. PMID- 3039800 TI - Changes in the 5-HT-binding sites in the cerebral cortex, striatum and hypothalamus of rats during ageing and under the effect of dopaminergic agents. AB - The changes in the 5-HT1-binding sites, developing during ageing and under the effect of 10-day administration of L-DOPA and haloperidol, were studied in the cerebral cortex, striatum and hypothalamus of 2-, 10- and 22-month-old rats. The values of Bmax of the 5-HT1-binding sites decreased in all three brain structures studied with the ageing of the experimental animals. Both L-DOPA and haloperidol did not change the general character of the changes in the number of 5-HT1 receptors, developing during ageing, but at the same time L-DOPA reduced their density in the three age periods and in the three cerebral areas studied, while haloperidol increased their number in the 2- and 10-month-old rats. The Kd-values of the 5-HT1-receptors manifested very varied changes, determined both by ageing and by the administration of L-DOPA and haloperidol. The generalized interpretation of these results with the results obtained in earlier studies on the age-determined changes in DA2 and the enkephalin receptors, as well as of the content of biogenic monoamines and MAO-activities, made it possible to see new elements in the adaptive possibilities of the cerebral receptors. On the other hand, the results obtained in the present and earlier studies show that ageing is paralleled by deviations in the equilibrium of the cerebral transmitter systems which seem to play an important role for the development of the behavioural changes connected with ageing. PMID- 3039801 TI - Leucine-enkephalin as a modulator of neurotransmission in cat lower esophageal sphincter. AB - Interactions between cholinergic, adrenergic and noncholinergic nonadrenergic inervation in lower esophageal sphincter determine the character of smooth muscle responses to field electrical stimulation. The neurogenic responses of muscle strips from the proximal (LES1), middle (LES2) and lower (LES3) parts of lower esophageal sphincter (LES) are: primary relaxation and secondary contraction in LES1, relaxation in LES2 and primary contraction followed by primary relaxation in LES3. Leucin-enkephalin in concentration of 0.1 microM and 1 microM leads to: increasing of amplitude of the primary relaxation and inhibition of secondary contraction in LES1, enhancing of amplitude of relaxation in LES2 and converts the response in LES3 from two into three component one so that the primary relaxation and the primary contraction are followed by secondary poststimulation relaxation. These effects are most probably the results of inhibition of noradrenaline and acetylcholine releasing under the influence of leucine enkephalin and of alteration of the correlation between the neurotransmitters. It is assumed that Leucine-enkephalin operates in LES as a neuromodulator inhibiting both the adrenergic and cholinergic neurotransmission. PMID- 3039802 TI - Sandfly fever epidemiology in Croatia. PMID- 3039803 TI - Malignant fibrous histiocytoma of the pericranium: case report. AB - Malignant fibrous histiocytoma only infrequently arises from structures of the head and neck. When it does, it most often originates in facial structures, particularly the maxilla. This case report details a malignant fibrous histiocytoma arising from the pericranium, and presenting clinically as a large indurated frontal calvarial mass. A CT scan showed its origin to be from the pericranium and demonstrated its intracranial extension. Radical excision presented a surgical challenge because of the extensiveness of the lesion. Yet, combined with post-operative teletherapy, a favourable outcome was obtained. PMID- 3039804 TI - [A case of metastatic cylindroma of the mandible or 10 years of diagnostic uncertainty]. PMID- 3039805 TI - [Local and loco-regional anesthesia in oral medicine in alcoholics]. PMID- 3039806 TI - The use of BHK suspension cells for the production of foot and mouth disease vaccines. PMID- 3039807 TI - Serum-free growth of human hepatoma cells. A review. PMID- 3039808 TI - Human papillomaviruses and genital cancer. PMID- 3039809 TI - Herpes simplex type 2 virus and cervical neoplasia. PMID- 3039810 TI - Transforming genes and target cells of murine spleen focus-forming viruses. PMID- 3039811 TI - Superoxide radicals (SR) in the pathophysiology of ischemic acute renal failure (ARF). PMID- 3039812 TI - Hepatitis A. PMID- 3039813 TI - Adeno-associated viruses: an update. PMID- 3039814 TI - [Change in rickets prevention and readjustment of the vitamin D regimen ("new" Dekristol)]. PMID- 3039815 TI - Decrease of beta-receptors in asthmatic and rhinitic patients. AB - The number of beta-receptors was determined in 60 atopic patients: 30 asthmatics with sensitization to domestic inhalant allergens and 30 rhinitic patients with sensitization to pollen and/or Dermatophagoides antigens. A L-iodocyanopindolol radio-active marker was used. Our control population (20 healthy individuals) showed 908 +/- 396 receptors/cell. The rhinitic individuals showed a 24% decrease in receptors in relation to the control population. On the other hand, within the asthmatic patients there existed 2 differentiated groups: inactive asthmatics whose receptors were found to be within the normal limits and active asthmatics with a 36% decrease in receptors in relation to the healthy individuals (673 receptors/cell) establishing a significant statistical difference (p less than 0.005). As a consequence these data seem to support Szentivanyi's B adrenoreceptor theory. PMID- 3039817 TI - Paget's disease of the breast. AB - Paget's disease of the breast is often an important early warning sign of underlying breast carcinoma, usually arising from the major mammary ducts. Because the conditions may be confused with benign diseases of the nipple, treatment is frequently delayed. If Paget's disease is the sole indication for surgery, the likelihood of cure is high. When evaluating nipple lesions, physicians should maintain a high index of suspicion for Paget's disease of the breast. PMID- 3039816 TI - [Effect of hyperlipemia on mitogen-induced blastogenesis in lymphocytes isolated from the rat spleen]. AB - Lipoproteins suppress several immune functions (mitogen-induced lymphoblast transformation, macrophage functions). Low density lipoprotein (LDL) is responsible for most of these effects. Diet may cause an elevation in LDL level. Therefore, we investigated the effect of cholesterol-rich diet upon the mitogen response of rats. The cholesterol and LDL-cholesterol level of rats was elevated after 8 days of diet. The blastogenic response of spleen cells separated by Ficoll-Paque gradient decreased. The response to phytomitogens 1, and 10 micrograms/ml phytohaemagglutinin (PHA) and 10 micrograms/ml concanavalin A (Con A) was measured by thymidine incorporation after 3 days of culture. The decrease of blastogenic response was more pronounced at the higher doses (10 micrograms/ml PHA, and 10 micrograms/ml Con A). For the lymphocyte cultures that did not contain autologous (rat) serum, we suggest that suppressed response was caused by an alteration of either in the membrane or in the metabolism of cells participating in the mitogen response. The decreased amount of LDL receptors and a derangement in intracellular cholesterol biosynthesis are supposed. The clinical implications of the immunosuppressive effect of dietary cholesterol is discussed. PMID- 3039818 TI - A laboratory for handling chemical toxicants safely. AB - The Centers for Disease Control has a special Chemical Toxicant Laboratory (CTL) for handling very hazardous chemicals. It is designed to protect the workers, prevent the release of the chemical toxicant into the surrounding environment, and provide for the scientific integrity of the experiments conducted. A discussion of laboratory ventilation and special containment devices is presented. The design of the CTL, coupled with a realistic set of safety guidelines, provides for the safe conduct of research involving highly toxic chemicals. PMID- 3039819 TI - The development and evaluation of a hydrobromic acid-coated sampling tube for measuring occupational exposures to ethylene oxide. AB - A new sampling method has been developed for the measurement of ethylene oxide (EtO) exposures in the workplace. This sampling method uses a hydrobromic acid coated charcoal tube to collect EtO as its 2-bromoethanol reaction product. Because 2-bromoethanol is less volatile and less reactive than EtO, improved sampling capacity and sample stability are observed with this method over the collection of EtO on untreated charcoal. Sample analysis is performed by gas chromatography with electron capture detection (GC/ECD) following desorption with dimethylformamide (DMF) and derivatization of the 2-bromoethanol reaction product with heptafluorobutyrylimidazole (HFBI). The direct analysis of 2-bromoethanol by GC/ECD was not reproducible and this effect was attributed to the presence of excess hydrobromic acid (HBr) in the sample. This matrix effect was eliminated by the formation of the heptafluorobutyrate ester, a water-insoluble derivative of 2 bromoethanol, which was extracted into iso-octane for analysis. This analytical scheme is very sensitive and was used to monitor test atmospheres of EtO at the 0.1 ppm level. An average recovery of 96% was obtained for short-term samples (15 minutes) collected from a 5-ppm test atmosphere at 23 degrees C and 80% relative humidity. Similar high recoveries were obtained for 4-hr samples collected at a sampling rate of 0.1 L/min from test atmospheres in the concentration range of 0.1 to 16 ppm EtO at high humidity (80%) and ambient temperature (22 degrees C to 25 degrees C). Samples collected under these same conditions and stored for a minimum of 2 weeks resulted in average recoveries that ranged from 84% to 101%. Average recoveries of 97% were obtained for 2-ppm air samples collected at low humidity with no storage; however, storage of these samples at 22 degrees C to 25 degrees C resulted in an approximate loss of 5% per week. A field comparison study of samples, that was obtained from a hospital sterilization facility, used the test method and the Qazi-Ketchum sample method and resulted in a correlation for paired samples of 0.99 over a concentration range of 0.3 to 7.0 ppm EtO. These results indicate that this sampling and analytical method is a convenient, accurate and reliable means of monitoring EtO exposures in the workplace. PMID- 3039820 TI - A sequential tape monitor for toluene diisocyanate. AB - An electronic microcircuit controller has been designed to convert a commercially available continuous tape monitor for toluene diisocyanate (TDI) into a sequential sampler in order to eliminate deficiencies in response and resolution of fluctuating TDI concentrations. The modified monitor will collect up to fourteen 12-min samples per work shift. The sensitivity of the monitor was doubled approximately by the inclusion of a blue filter in the optics of the reader unit. Recalibration of several modified monitors indicated a wide range in response to TDI necessitating individual calibrations. No significant difference in response to the 2,4 and 2,6 isomers of toluene diisocyanate was noted for modified personal monitors, in contrast to a 25% lower response to 2,6-TDI by an area monitor in which the same detection tape was used. The modified monitor has shown a reliability rate of about 90% during a three-year field survey of TDI exposures. PMID- 3039821 TI - Toluene diisocyanate exposures in the flexible polyurethane foam industry. AB - A 3-year survey of toluene diisocyanate exposure in two flexible polyurethane foam manufacturing facilities has been conducted. The geometric mean time weighted average exposures were 2.36 ppb, 1.10 ppb, and 1.50 ppb for the foam line workers, finishing workers, and maintenance personnel, respectively. The OSHA ceiling standard of 20 ppb was exceeded by 1.3% of the short-term (12 min) exposure measurements taken. Exposures were shown to be predominantly to the 2,6 isomer of TDI. PMID- 3039822 TI - The effect of electrostatic charge on the aspiration efficiencies of airborne dust samplers: with special reference to asbestos. AB - An experimental investigation has been conducted into the effects of electrostatic charge, carried by the dust particles and by the sampler itself, on the sampling of airborne dusts. Experiments covering both personal and static sampling and a range of sampler types were carried out in the laboratory for both fibrous asbestos and isometric silica gel dusts. Experiments also were carried out in the spinning shop of an asbestos textile factory. The results showed that the aspiration efficiency of the sampler always is reduced as the charge on the sampler increases, independently of the type of sampler and of whether it is used as a static or personal sampler. The effect is most marked when sampling takes place in calm air. It is concluded from the results that, for the levels of charge reached by samplers in most practical situations, the effects on aspiration efficiency will be small. Possible exceptions to this might occur, however, in workplace environments where relative humidity is very low, and charge levels of the sampler (or on the worker wearing the sampler) can become high. PMID- 3039823 TI - Cytoplasmic bar-like structures of alveolar type II cells: an ultrastructural study in freshly isolated cells from rat lungs. AB - Bar-like structures are tubular cytoplasmic inclusions found in situ in pulmonary epithelial type II cells of several animal species. The physiological significance and mode of formation of these inclusions are not fully established. In this paper, we describe bar-like structures as found in freshly isolated type II cells from rat lungs. Pulmonary cells were dissociated from the tissue with elastase and separated on a discontinuous density gradient of Percoll. The complete isolation procedure yielded 17 X 10(6) type II cells per animal (purity = 80%). Either from the crude cell suspensions or the purified preparations, only a small fraction of the type II cell population harbored the inclusions (less than 5%). It is shown that the bounding membranes of the bar-like structures can derive from the endoplasmic reticulum, the nuclear membrane, or the Golgi apparatus. Occasional connections with lamellar bodies were observed, and different levels of complexity in the bar-like structures were also found. The apparent rigid conformation and the orientation of the bar-like structures were taken as evidence for a role of the cytoskeleton in their formation. Because the inclusions do not appear to be new organelles or cellular structures performing a specific function, we propose that their formation may be a transient and limited cellular event in normal cells. However, the stabilization and the generation of the osmiophilic structures, as well as their overproduction, may reflect alterations of the normal physiology of the type II cells. PMID- 3039824 TI - Tridimensional architecture of the Golgi apparatus and its components in mucous cells of Brunner's glands of the mouse. AB - The three-dimensional structure of the Golgi apparatus and its components has been analyzed in thin and thick sections of mucous cells of mouse Brunner's glands by using low- and high-voltage electron microscopes and a stereoscopic approach. In thick sections of glands impregnated with osmium or treated to detect nicotinamide adenine dinucleotide phosphatase (NADPase) or thiamine pyrophosphatase (TPPase) activity, the Golgi apparatus appeared, at low magnification, as a continuous network located in the supranuclear region. At higher magnifications and in thin sections of tissue postfixed with reduced osmium and stained with lead citrate or treated to demonstrate phosphatase activity, the following components were observed: on the cis-face of the Golgi stacks, an osmiophilic tubular network referred to as the cis-element; a cis saccular-compartment composed of a distended porous saccule slightly reactive for NADPase and three or four underlying NADPase-positive, flattened, poorly fenestrated saccules; a trans-saccular-compartment consisting of four to six TPPase-positive saccules or sacculo-tubular elements, prosecretory granules, and "peeling off" trans-tubular networks. The saccules of the cis-compartment were often perforated by large pores in register. The cavities thus formed in the stacks were called wells and were pan-shaped with a mouth directed toward the cis face of the stacks and a bottom closed by TPPase-positive saccules. The wells always contained 80-nm vesicles. The saccules of the trans-compartment were involved in the formation of secretory granules according to the following proposed sequence of transformation. The secretion product appeared initially as a granular material evenly distributed throughout a slightly distended, poorly fenestrated saccule. These saccules appeared to transform into fenestrated elements with irregular pores and with parts of them taking on the appearance of a tubular network; they were thus referred to as sacculotubular elements. The secretory material initially distributed throughout these elements accumulated in nodular dilatations randomly distributed along the tubular portions of the elements. The dilatations, considered as prosecretory granules, increased in size as they drained the secretory material from the rest of the sacculotubular elements. Such prosecretory granules, large and irregular in shape, "peeled off" from the stacks of saccules with residual saccular or tubular structures still attached to them, some of the latter forming trans-tubular networks. The prosecretory granules detached from such membranous residues, condensed, and finally transformed into spherical secretion granules. PMID- 3039825 TI - Folate conjugase activity in the plasma and tumors of breast-cancer patients. AB - The activity of folate conjugase (pteroylpolyglutamate hydrolase, EC 3.4.22.12) was measured in plasma from normal subjects and patients with breast cancer using pteroylglutamyl-gamma-glutamyl-gamma-(U14C) glutamate as the substrate. Conjugase assays also were performed on samples of breast-tumor tissue, normal breast, and fibroadenoma. When assayed at pH 4.5 and 7.4, mean plasma conjugase activity was significantly (p less than 0.05) elevated in a group of patients with anatomically proven metastatic disease (n = 21) when compared with control subjects (n = 12) and a group of patients (n = 13) with no clinical evidence of disease after mastectomies. Mean plasma conjugase activity assayed at pH 4.5 also was significantly higher in the metastatic disease group when compared with breast cancer patients before mastectomy (n = 9) and fibroadenoma patients before biopsy (n = 3). The specific activity of tissue conjugase assayed at pH 4.5 was significantly higher (p less than or equal to 0.05) in infiltrating ductal carcinoma than in normal adjacent breast tissue according to the Wilcoxon test for paired samples (n = 10). PMID- 3039826 TI - Mineral balance in adult men: effect of four refined fibers. AB - Eleven men consumed a basal diet alone and with cellulose (Na carboxymethylcellulose, locust bean gum, or karaya gum) added at 7.5 g fiber per 1000 calories for 4 wk each. Food, urine, and fecal composites were collected during the last 8 d of each feeding period. Bowel transit time was not significantly affected; however, total dry fecal weight was significantly increased after the refined fibers compared with that after the basal diet. Adding refined fibers to the basal diet did not significantly affect apparent mineral balance of calcium, magnesium, manganese, iron, copper, or zinc, with the exception of a negative mineral balance for manganese with carboxymethylcellulose. Karaya gum had a mean positive balance for all minerals tested. These results indicate that the hypocholesterolemic effect of the fibers that form gels occurs without compromising mineral balance in those subjects consuming Recommended Dietary Allowance levels of the minerals studied. PMID- 3039827 TI - Omega-3 fatty acid requirement. PMID- 3039829 TI - CT-guided stereotactic biopsy of brain tumors: pathologic considerations. AB - Astrocytic tumors, the most common of the primary intracerebral neoplasms, are a heterogeneous group in terms of clinical behavior and pathologic appearance. Current histopathologic classification emphasizes this aspect; however, its application to tissue specimens removed by means of CT-guided stereotactic brain biopsy requires special considerations. Pertinent aspects of pathology relevant to assessment by CT scan stereotactic needle biopsy are discussed, together with a presentation of early results on the use of this diagnostic technique. PMID- 3039828 TI - Growth of astrocytomas in the human tumor clonogenic assay and sensitivity to mismatched dsRNA and interferons. AB - Nine astrocytoma specimens were received from seven patients and processed for testing in the human tumor clonogenic assay (HTCA). Cells derived from these specimens were challenged with human natural alpha-interferon (alpha-IFN) and beta interferon (beta-IFN), recombinant beta interferon (beta ser-IFN), and mismatched double-stranded (ds) RNA (Ampligen). Six of the astrocytoma specimens formed adequate colonies for drug sensitivity testing (greater than or equal to 30 colonies/plate), and all were high-grade (III-IV) tumors. Sensitivity was defined as a greater than or equal to 50% decrease in tumor colony formation following drug exposure and was observed with alpha-IFN (2/4), beta-IFN (3/4), and mismatched dsRNA (4/5) exposure. No decrease in colony growth was observed after recombinant beta ser-IFN exposure, and in 2 of 3 cases, colony formation was stimulated. The sensitivity of 75 non-CNS solid tumors to mismatched dsRNA was compared to the high-grade astrocytomas in the HTCA. Of the 10 additional histologic tumor types studied, carcinoid and renal cell carcinomas exhibited the greatest sensitivity to mismatched dsRNA: 63% and 52%, respectively. However, in comparison, 80% of the high-grade astrocytomas were sensitive, demonstrating that these gliomas are among the most sensitive of human tumors to mismatched dsRNA in vitro. Clinical trials of interferons and mismatched dsRNA, coupled with in vitro sensitivity studies, should further define their therapeutic potential. PMID- 3039830 TI - Primary malignant mediastinal germ-cell tumors and the contribution of radiotherapy: a southeastern multi-institutional study. AB - A retrospective review was performed by a multi-institutional study group to determine the contribution of radiotherapy to the management of primary malignant mediastinal germ-cell tumors. Twenty-seven patients diagnosed with a primary mediastinal germ-cell tumor between January 1965 and July 1985 form the basis of this study. Twenty-five of the 27 patients were male. Thirteen patients' tumors were diagnosed as seminoma and the remaining 14 patients' tumors had other germ cell histologies. The single most important prognostic factor was histology, with a 5-year actuarial survival of 100% for the seminomas and only 8.8% for the remaining germ-cell varieties. If total surgical extirpation is not possible, biopsy may be adequate. Of the patients with seminoma, 11 of 12 had local control, and 3 of the 12 patients were treated with doses between 3,000 and 3,100 cGy. High doses for this variety of mediastinal germ-cell tumor might not be required. For the germ-cell tumors other than seminoma, no patient had local control with doses over the range of 3,000-4,750 cGy. PMID- 3039831 TI - Antiemetic efficacy of nabilone and dexamethasone: a randomized study of patients with lung cancer receiving chemotherapy. AB - In a previous study on the antiemetic effect of nabilone (N) in patients with lung cancer receiving chemotherapy (CT), we found that N was only moderately effective and that its side effects limited its use, especially in elderly outpatients. We, therefore, performed a new study of N in combination with dexamethasone (DXM), a potent antiemetic in itself, to evaluate whether the addition of DXM to N would improve the antiemetic effect and/or reduce the side effects. Forty patients with lung cancer were enrolled in the study. A randomized, third-party-blinded, crossover design was used. Study drugs were given during two consecutive, identical CT cycles. N was given at a fixed dosage regimen of 2 mg b.i.d. The initial dose was administered the evening before CT, the second dose at 0.5 h before CT, and the third dose in the evening 12 h after CT. DXM, 8 mg, or placebo was given orally with the first dose of N. The subsequent doses (either 10 mg DXM or saline) were given intravenously 0.5 h before CT and at 2 and 6 h after the start of CT. The CT regimens given included the following drugs in various combinations: cisplatin, cyclophosphamide, adriamycin, etoposide (VP-16), vincristine, and vindesine. The combination of N and DXM was significantly superior to N alone in the reduction of vomiting episodes, both in subgroups of patients receiving cisplatin and in those receiving other CT combinations. There was no statistically significant difference between the treatments with regard to the patients' assessments of the severity of nausea or effects on appetite. Approximately half the patients (63% with N plus DXM versus 47% with N) reported no side effects. The frequency and severity of central nervous system adverse reactions, mainly vertigo, were similar in both treatment groups. The fall in blood pressure was significantly greater after N alone. Two thirds of the patients preferred N plus DXM. Thus, the addition of DXM to N enhanced the therapeutic yield of N, and we recommend DXM as an adjunct to N, when the use of steroids is not contraindicated. The optimal dose and schedule of DXM was not investigated in our study; a higher dose of DXM might increase the clinical benefit of the drug combination tested. PMID- 3039832 TI - Nuclear DNA content in squamous cell carcinoma, small cell carcinoma, and bronchiolo-alveolar carcinoma of the lung. AB - Nuclear DNA content in individual, morphologically identified tumor cells from 33 squamous lung carcinomas, 20 small cell lung carcinomas, and 10 bronchiolo alveolar carcinomas were analyzed by means of cytophotometry on Feulgen-stained histologic and cytologic specimens. Twenty-eight of the squamous cell carcinomas and 17 of the small cell carcinomas had high and scattered DNA values, indicative of high malignancy potentials. None of the bronchiolo-alveolar carcinomas showed such high DNA values. These results are in line with clinical experience that squamous cell and small cell carcinoma are associated with rapid progression and death in patients, whereas bronchiolo-alveolar carcinomas have a more indolent course. PMID- 3039833 TI - Cerebral venous sinus thrombosis in a patient receiving adjuvant chemotherapy for stage II breast cancer through an implanted central venous catheter. PMID- 3039834 TI - Esthesioneuroblastoma. Intermediate filaments, neuroendocrine, and tissue specific antigens. AB - Esthesioneuroblastoma (EN), a malignant neuroblastic tumor arising in the superior portion of the nasal cavity, shares histologic similarities with a number of primary malignant tumors that arise in this region, including rhabdomyosarcoma, lymphoepithelioma, and lymphoma. To establish an antigenic profile of EN as an aid in the differential diagnosis of these histologically similar nasal tumors, immunostaining was performed for the following intermediate filaments: keratin, neurofilament, glial fibrillary acidic protein, and desmin; neuron-specific enolase (NSE), S-100 protein, chromogranin, human common leukocyte antigen (HLE), epithelial membrane antigen (EMA), myoglobin, and carcinoembryonic antigen (CEA) on 21 primary nasal tumors: eight EN, five lymphoepitheliomas, two small cell carcinomas, three lymphomas, and three rhabdomyosarcomas. Keratin and CEA stained only the carcinomas (6/7+, 4/7+), respectively; desmin and myoglobin only rhabdomyosarcoma (3/3+, 1/3+); and HLE only lymphomas (3/3+). Chromogranin and neurofilament staining occurred exclusively in one case each of EN. S-100 and NSE commonly stained EN (5/8+, 6/8+), but carcinomas (1/7+, 2/7+) and rhabdomyosarcomas (1/3+, 3/3+) were also positive. Despite the apparent nonspecificity of NSE and S-100, an antigenic profile of positive NSE of S-100 staining with negative epithelial, muscle, and lymphoid antigens uniquely identified six of eight EN. Chromogranin and neurofilament positivity was further evidence for EN in two cases. This antigenic profile is a helpful adjunct in the diagnosis of EN and other primary malignant nasal tumors. PMID- 3039835 TI - Increased expression of N-myc in human small cell lung cancer biopsies predicts lack of response to chemotherapy and poor prognosis. AB - Amplified and increased expression of the myc family of protooncogenes (c- and N myc) has been described to be associated with rapid proliferation in a number of cell lines, including small cell lung cancer (SCLC). In SCLC, c-myc was demonstrated to be amplified in a subset of SCLC cell lines denoted as variant type, which show a more aggressive way of growth in vitro. The N-myc oncogene, which has extensive homology in the second exon with c-myc, has been shown to be implicated in the oncogenesis of several primary tumors, including SCLC. The authors describe, using in situ hybridization, that increased expression of the N myc oncogenes in primary biopsies from 15 untreated patients with SCLC are strongly associated with poor response to chemotherapy, rapid tumor growth, and short survival. PMID- 3039836 TI - Clear cell carcinoma arising in pleomorphic adenoma of the salivary gland. AB - Malignant change in pleomorphic adenoma of the salivary gland is not common. Clear cell carcinoma is an extremely rare form of malignant salivary gland tumor. A case of clear cell carcinoma of the submandibular gland arising in pleomorphic adenoma is reported. PMID- 3039837 TI - Malignant granular lymphoproliferation after Epstein-Barr virus infection: partial immunologic reconstitution with interleukin-2. AB - This report describes a patient who developed a malignant proliferation of granular lymphocytes following Epstein-Barr virus (EBV) infection. For many months, his illness resembled prolonged infectious mononucleosis with persistent fatigue, fever, leukocytosis, and serologic evidence of recent primary EBV infection. After approximately 1 year, however, he developed progressive granular lymphocytosis and extensive lymphocytic infiltration of the bone marrow and liver. Tests for EBV DNA in pre- and postmortem tissue samples using a sensitive DNA hybridization technique were negative. Southern blot analysis of DNA prepared from blood mononuclear cells demonstrated clonal T-cell antigen receptor gene rearrangement. Despite increased numbers of circulating lymphocytes with the morphology and surface phenotype of normal donor natural killer (NK) cells, the patient's NK activity was consistently depressed in a standard in vitro assay. However, in vitro incubation with interleukin-2 (IL-2), but not with alpha- or gamma-interferon, increased the NK activity of the patient's lymphocytes. Intravenous recombinant IL-2 treatment transiently increased the patient's blood NK activity and was associated with seroconversion to EBV nuclear antigens but failed to affect the progression of his disease. Our findings indicate that clonal granular lymphocytic proliferation may develop after EBV infection and confirm the utility of DNA hybridization analysis in distinguishing monoclonal from benign immunoreactive lymphoproliferation. Furthermore, our results suggest that certain functionally inert neoplastic granular lymphocytes acquire NK activity when exposed to IL-2. PMID- 3039838 TI - A unique dicentric X;Y translocation with Xq and Yp breakpoints: cytogenetic and molecular studies. AB - A 32-year-old woman presented with secondary amenorrhea and infertility. She was of normal height and her breasts were well developed, but she had streak gonads; there were no signs of virilization, and she showed no somatic stigmata of Turner syndrome. Chromosome analysis revealed a dicentric X;Y translocation with Xq and Yp breakpoints. Centromeric banding demonstrated a Y centromere and a "suppressed" X centromere. The karyotype of the patient was interpreted as 46,X,t(X;Y)(q22;p11). The Yp breakpoint was confirmed by DNA-hybridization studies with six probes detecting Y-specific sequences. These DNA-hybridization studies were consistent with the presence of the long arm, centromere, and much of the proximal short arm of the Y. The Y-DNA studies of this female also revealed the absence of the distal short arm of the Y chromosome, to which the testis-determining factor has previously been localized. PMID- 3039840 TI - Tremolite/chrysotile ratios. PMID- 3039839 TI - Genetic mechanisms of tumor-specific loss of 11p DNA sequences in Wilms tumor. AB - Wilms tumor, a common childhood renal tumor, occurs in both a heritable and a nonheritable form. The heritable form may occasionally be attributed to a chromosome deletion at 11p13, and tumors from patients with normal constitutional chromosomes often show deletion or rearrangement of 11p13. It has been suggested that a germinal or somatic mutation may occur on one chromosome 11 and predispose to Wilms tumor and that a subsequent somatic genetic event on the normal homologue at 11p13 may permit tumor development. To study the frequency and mechanism of such tumor-specific genetic events, we have examined the karyotype and chromosome 11 genotype of normal and tumor tissues from 13 childhood renal tumor patients with different histologic tumor types and associated clinical conditions. Tumors of eight of the 12 Wilms tumor patients, including all viable tumors examined directly, show molecular evidence of loss of 11p DNA sequences by somatic recombination (four cases), chromosome loss (two cases), and recombination (two cases) or chromosome loss and duplication. One malignant rhabdoid tumor in a patient heterozygous for multiple 11p markers did not show any tumor-specific 11p alteration. These findings confirm the critical role of 11p sequences in Wilms tumor development and reveal that mitotic recombination may be the most frequent mechanism by which tumors develop. PMID- 3039841 TI - Ganciclovir for the treatment and suppression of serious infections caused by cytomegalovirus. AB - Ganciclovir is a congener of acyclovir with in vitro activity against cytomegalovirus. Ninety-seven patients with the acquired immune deficiency syndrome (AIDS) and a serious cytomegalovirus infection received ganciclovir, 3.0 to 15 mg/kg per day. Viremia cleared during drug therapy in 88 percent of patients. Viral shedding from urine and throat ceased or became inapparent during treatment in 78 percent and 68 percent of patients, respectively. Among patients with cytomegalovirus retinitis, 87 percent of evaluable patients had improvement in (30 of 60) or stabilization (22 of 60) of their disease. However, when the drug was discontinued, progression or recurrence of disease always occurred. Long term suppressive therapy with ganciclovir, 5.0 mg/kg five to seven times weekly, prevented the recurrence of cytomegalovirus disease (p less than 0.001). The drug was eliminated by renal excretion, and in patients without renal impairment (creatinine clearance rates of more than 60 ml/minute/1.73 m2), ganciclovir has a mean half-life of 4.2 hours. Significant neutropenia and leukopenia occurred in 55 percent and 32 percent of patients, respectively. PMID- 3039842 TI - Treatment of mild to moderate hypertension with or without the converting enzyme inhibitor enalapril. Results of a six-month double-blind trial. AB - A group of 205 patients with mild to moderate essential uncomplicated hypertension was chosen from 3,183 hypertensive patients referred to a hypertension clinic for the first time, and was asked to participate in a six month double-blind parallel trial. A single physician was in charge of all the patients. After a two-week single-blind placebo period, the patients were randomly assigned to regimens of either enalapril (20 mg per day) or placebo. Both groups were then followed up every two weeks, and increasing doses of hydrochlorothiazide (25 and 50 mg), oxprenolol (160 and 320 mg), and dihydralazine (50 and 100 mg) were added until the diastolic blood pressure was lower than 90 mm Hg. After a six-month follow-up, the enalapril group showed lower systolic and diastolic blood pressures than the control group (129/82 +/- 12/6 mg Hg versus 135/86 +/- 10/5 mm Hg; p less than 0.001). The number of daily tablets of active drugs was 2.7 +/- 1.8 in the enalapril group and 4.4 +/- 2.4 in the control group (p less than 0.01). The mean plasma potassium level was 4.16 +/ 0.4 mmol/liter in the enalapril group versus 3.92 +/- 0.4 mmol/liter in the control group (p less than 0.001), despite more frequent use of amiloride (p less than 0.001). This difference is explained by the lower dose of hydrochlorothiazide used in the enalapril group by comparison to the control group, and a lower excretion of urinary aldosterone in the enalapril group than in the control group (11.6 +/- 7.4 versus 19.8 +/- 11.8 micrograms per 24 hours, p less than 0.001). Drug withdrawal was necessary in eight patients in the enalapril group and in 16 patients in the control group (p less than 0.05). These results show that first-step treatment of mild to moderate uncomplicated essential hypertension with enalapril permits better blood pressure control than the standard treatment, requires fewer tablets to be taken daily, and involves a smaller risk of hypokalemia. PMID- 3039843 TI - Serum angiotensin converting enzyme activity in evaluation of patients with liver disease. AB - This study was undertaken to evaluate angiotensin converting enzyme activity in patients with liver disease, and to explore its relationship to thyroid function in patients with liver disease. Serum angiotensin converting enzyme activity and thyroid hormone levels, determined in 79 patients with intrahepatic-type liver disease and 18 patients with extrahepatic biliary obstruction, were compared with values in 129 euthyroid controls. Serum angiotensin converting enzyme activity was higher in intrahepatic disease than in extrahepatic obstruction, but statistically significant only for acute intrahepatic disease. In the patients studied, high serum angiotensin converting enzyme activity (22 units/ml or more) virtually excluded extrahepatic biliary obstruction, with a negative predictive value of 94 percent. Low serum angiotensin converting enzyme activity had a positive predictive value for extrahepatic obstruction of only 67 percent. Increased angiotensin converting enzyme activity could not be explained as a function of enhanced thyroid activity. Serum angiotensin converting enzyme activity may be useful in separating patients with intrahepatic liver disease from those with extrahepatic obstruction. PMID- 3039844 TI - Control of gastric secretion. University of California/Davis Interdepartamental Conference. PMID- 3039845 TI - Varicella zoster antibody testing in the care of pregnant women exposed to varicella. AB - Appropriate use of varicella zoster immunoglobulin to prevent or ameliorate maternal or perinatal infection depends on accurate identification of varicella susceptible women. Detection of fluorescent antibody to varicella zoster virus membrane antigen is an available means for identifying women exposed to varicella who are at risk for infection. During a 5 1/2-year period, 37 pregnant women who had been exposed to varicella and who had a negative or indeterminate history of prior varicella were tested for the presence of varicella zoster virus membrane antigen. Among the 17 women with a negative history, 12 (71%) had a positive test result and five (29%) had a negative test result. Among 20 women with undeterminant histories, 18 (90%) were immune and two (10%) were susceptible. Three additional women who were tested despite a history of varicella had positive varicella zoster virus membrane antigen test results. Overall, 81% of patients with a negative or uncertain history of varicella had serologic evidence of past varicella. One of six untreated seronegative women and one weakly positive woman developed disease. There was no instance of congenital varicella. Expeditious determination of varicella zoster virus membrane antigen or equivalent anti-varicella antibody status in pregnant women exposed to varicella appears to be a rapid, satisfactory method for determining who should promptly receive varicella zoster immunoglobulin passive immunization. PMID- 3039846 TI - Tamoxifen retinopathy at low dosage. PMID- 3039847 TI - Inhibition of epidermal growth factor-induced cellular proliferation. AB - The authors tested whether two oncogenic poxviruses, Shope fibroma virus (SFV) and malignant rabbit fibroma virus (MV), coded for an epidermal growth factor (EGF)-like protein. Virus-free lysates of SFV or MV-infected rabbit kidney cells do not appreciably affect proliferation of EGF-unresponsive RK-13 cells. Comparable lysates inhibit the background proliferation of two EGF-responsive cell lines, human foreskin fibroblasts and normal rat kidney cells. Inhibition of EGF-stimulated proliferation is also observed in a dose-dependent fashion. This inhibition is greatest when the virus lysate preparations are added either simultaneously with or before EGF. When EGF is added to cultures 24 hours before the SFV or MV preparations, the latter then have little or no effect on EGF induced target cell proliferation. The inhibitory factor competed with radiolabeled EGF for its receptor site. Electroblotting shows a protein of 35 kd molecular weight in the lysates of virus-infected RK-13 cells which reacts with anti-EGF antibody. These findings interpreted to indicate that SFV and MV code for an inhibitor of EGF activity, and that this inhibition occurs at least in part by competitive inhibition of EGF-EGF receptor interactions. PMID- 3039848 TI - Immunohistochemical study of basement membrane antigens in bronchioloalveolar carcinoma. AB - Bronchioloalveolar carcinoma (BAC), not yet completely defined as a biologic entity, has recently been classified into two different types. Immunohistochemical investigations, aimed at characterizing basement membrane (BM) behavior in the two types of BAC, revealed different distribution patterns. The first (Type I BAC) showed a linear staining for laminin and Type IV collagen similar to normal lung. Fibronectin was widely present in the septal interstitium and patchily distributed along the BM. The second (Type II BAC) showed a variable reaction for Type IV collagen and fibronectin, whereas laminin was absent or appeared as short, interrupted tracts around the epithelial neoplastic population, similar to conventional adenocarcinoma of the lung. These results suggest that only Type I BAC shows structural characteristics different from those of conventional adenocarcinoma of the lung. PMID- 3039849 TI - Aortic endothelial cell proteoheparan sulfate. I. Isolation and characterization of plasmamembrane-associated and extracellular species. AB - Proteoheparan sulfate biosynthesis was studied in cultured bovine aortic endothelial cells by means of pulse and pulse-chase experiments and subcellular fractionations. Three proteoheparan sulfate species were found in the medium. The major species, which the authors have called HS I, appeared in the medium only after an initial lag period and was also found associated with the plasma membrane. The other two (HS II and HS III) appeared in small amounts in the medium at early time points. At later times these were not readily observed because the large amounts of HS I present in the medium. The major medium species, HS I, appeared to be composed of approximately four heparan sulfate chains of approximately 35,000 daltons and a core protein of approximately 55,000 daltons apparent molecular weight. HS I appeared to be homogeneous by gel filtration on Sepharose CL 2B and 6B and elution from DEAE Sephacel, electrophoresis on Nu-Sieve agarose, and CsCl density centrifugation. After digestion with heparinase the core protein appeared to be homogeneous by S-200 Sephacel chromatography. HS I was also found associated with plasma membrane fractions of the cultured bovine aortic endothelial cells, and antisera raised against it stained epithelial and endothelial cells in patterns consistent with a cell surface localization. Of the other two species found in the medium, one (HS II) also appeared to be a component of the cell layer. This species appeared to contain approximately four heparan sulfate chains of approximately 20,000 daltons apparent molecular weight. Antisera raised against a similar molecule produced by HR 9 cell cultures stained basement membranes intensely, supporting the subcellular matrix localization of this molecule. The third species (HS III) was detected in culture medium only and apparently contained two heparan sulfate chains of approximately 20,000 daltons apparent molecular weight. These results support the concept of multiple endothelial cell proteoheparan sulfate species which exhibit differences in structure and localization and possibly diverse specialized functions. PMID- 3039851 TI - Malignant neoplasms of decidual origin (deciduosarcomas) induced by estrogen progestin-releasing intravaginal devices in rabbits. AB - A combination of estrogen and levonorgestrel was continuously delivered to 23 adult rabbits for up to 2 years via a Silastic ring device sutured into the vagina. Twenty-one control rabbits were given similar rings devoid of drugs. A marked decidual reaction of the endometrium occurred in 16 of 23 test rabbits. In 14 test rabbits (61%) malignant tumors developed of decidual type cells not heretofore described. The deciduosarcomas were composed of anaplastic cells that invaded the uterine walls, uterine lymphatics, and in 4 of 13 (31%) rabbits that survived 2 years of treatment, the tumors metastasized to the lungs. Several deciduosarcomas appeared to arise within the spleen or other abdominal organs. Other drug-related lesions included uterine or vaginal polyps, endometrial atrophy, and focal necrosis and mineralization of the uterine wall. Cells from several deciduosarcomas failed to produce tumors in nude mice or to colonize on soft agar. No decidualization or decidual neoplasms were seen in the controls. PMID- 3039850 TI - Correction of a developmental defect in neutrophil activation and movement. AB - In order to more fully understand the mechanisms involved in the developmental defect in polymorphonuclear leukocyte (PMN) movement in human neonates, the authors have examined several events in the activation response sequence. Chemotactic factor receptor numbers have been found to be normal on the PMNs of neonates, but chemotactic factor-induced changes in membrane potential and cyclic adenosine monophosphate concentrations were markedly decreased to absent in the neonatal cells. Because the neonatal PMN lacks the ability to deform normally, we examined the effects of a methylxanthine derivative, pentoxifylline, on the responses of neonatal cells. This agent has been reported to increase cell deformability and improve cell movement. Pentoxifylline had an effect in improving chemotactic function in the PMNs of neonates, while correcting the abnormality in membrane potential. In addition, this agent was found to enhance the movement of cell surface concanavalin A receptors after colchicine treatment. These results suggest that this developmental defect in cell activation and movement may be an abnormality that can be corrected pharmacologically. PMID- 3039852 TI - Na-H antiport in cultured rat aortic smooth muscle: its role in cytoplasmic pH regulation. AB - We have investigated the role of the Na-H antiport in the regulation of intracellular pH (pHi) in vascular smooth muscle. Experiments were conducted on contractile-state rat aortic smooth muscle cells grown in primary culture and loaded with the pH-sensitive, fluorescent indicator 2',7',-biscarboxyethyl-5(6) carboxyfluorescein (BCECF). Cells equilibrated in a normal physiological salt solution (PSS) containing 135 mM Na, pH 7.4 at 37 degrees C, had a pHi of 7.16 +/ 0.04 (means +/- SE; n = 8). 5-(N-ethyl-N-isopropyl)amiloride (EIPA) caused a concentration-dependent fall in pHi. Removal of extracellular Na caused an intracellular acidification that was rapidly reversed on replacement of Na. The rate of recovery from NH4Cl-induced intracellular acidosis was dependent on extracellular Na concentration (Km 14.6 +/- 2.8 mM) and was accelerated by increasing the transmembrane Na gradient and slowed by decreasing it. Recovery from acidosis was completely abolished by either EIPA or the absence of extracellular Na. These results demonstrate that the Na-H antiport is an important mechanism for the maintenance and regulation of pHi in vascular smooth muscle cells. The BCECF fluorescence technique provides an ideal method for further studies on the mechanisms for pHi regulation in these cells. PMID- 3039853 TI - Mitogen stimulation of Na+-H+ exchange: differential involvement of protein kinase C. AB - The mitogen-induced activation of Na+-H+ exchange was investigated in two cultured human fibroblast strains (HSWP and WI-38 cells) that, based on previous studies, differed in their response to the tumor-promoting phorbol ester 12-O tetradecanoylphorbol-13-acetate (TPA) (L. M. Vincentini and M. L. Villereal, Proc. Natl. Acad. Sci. USA 82: 8053-8056, 1985). The role of protein kinase C in the activation of Na+-H+ exchange was investigated by comparing the effects of TPA on Na+ influx, in vitro phosphorylation, and in vivo phosphorylation in both cell types. Although both cell types have significant quantities of protein kinase C activity that can be activated by TPA in intact cells, the addition of TPA to intact cells stimulates Na+ influx in WI-38 cells but not in HSWP cells, indicating that in HSWP cells the stimulation of protein kinase C is not sufficient to activate the Na+-H+ exchanger. Cells were then depleted of protein kinase C activity by chronic treatment with high doses of TPA. Both HSWP and WI 38 cells were rendered protein kinase C deficient by this treatment as determined by in vitro and in vivo phosphorylation studies. Protein kinase C-deficient HSWP cells lose the ability for TPA to inhibit the serum-induced activation of Na+-H+ exchange, but there is no reduction in the stimulation of Na+ influx by serum, bradykinin, vasopressin, melittin, or vanadate, indicating that protein kinase C activity is not necessary for the mitogen-induced activation of Na+-H+ exchange in HSWP cells by agents known to stimulate phosphatidylinositol turnover (G. A. Jamieson and M. Villereal. Arch. Biochem. Biophys. 252: 478-486, 1987). In contrast, depletion of protein kinase C activity in WI-38 cells significantly reduces both the TPA- and the serum-induced activation of the Na+-H+ exchange system, suggesting that protein kinase C activity is necessary for at least a portion of the mitogen-induced activation of the Na+-H+ exchanger in WI-38 cells. These results indicate that the mechanisms for regulating Na+-H+ exchange can differ dramatically between different types of fibroblasts. PMID- 3039854 TI - Coupling of aerobic glycolysis and Na+-K+-ATPase in renal cell line MDCK. AB - The relation between the activity of the Na+-K+-ATPase and the metabolic source of ATP was investigated in suspensions of MDCK cells. The pump activity of Na+-K+ ATPase was estimated from the initial rate of ouabain-sensitive K+ uptake into K+ depleted cells. Uptake was initiated by the reintroduction of K+ to the medium in which the cells were suspended. The metabolic source of ATP was varied by changing the substrates supplied to the suspension. Cells respiring on glutamine produced ATP from oxidative metabolism alone, whereas cells incubated with glucose and glutamine produced ATP via glycolysis and oxidative phosphorylation. Over a wide range of extracellular K+ concentrations, the initial rate of K+ uptake was faster in cells incubated with glucose and glutamine when compared with cells incubated with glutamine alone. Kinetic analysis together with ouabain binding data demonstrated that this increase in K+ uptake was due to an increase in maximal velocity (Vmax) at a constant number of Na+-K+-ATPase transport sites. In addition, steady-state studies revealed that the addition of glucose to K+ depleted cells respiring on glutamine alone resulted in a net ouabain-sensitive influx of K+. These data demonstrate that in MDCK cells the maximal capacity for transport via the Na+-K+-ATPase is greater when ATP is produced from both glycolysis and oxidative phosphorylation than when ATP is produced from oxidative phosphorylation alone.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3039855 TI - Three distinct cell populations in rat kidney collecting duct. AB - The morphologically heterogeneous cell populations in the collecting ducts of the rat kidney were studied using immunocytochemical detection of Na+-K+-ATPase and the anion channel (band 3) glycoprotein. Both enzymes were localized to the basal aspect of separate and morphologically distinct subpopulations of cells in various segments of the collecting duct. Na+-K+-ATPase appeared to be present exclusively in principal cells as identified by their morphology, whereas band 3 antibodies reacted only with intercalated cells. However, 5-20% of cells with the morphological characteristics of intercalated cells failed to react with either antisera in various segments of collecting ducts. As band 3 glycoprotein serves in exchanging intracellular bicarbonate for chloride, it is highly likely that the cells positive for this antigen secrete protons. The method introduced here appears thus useful for distinguishing between principal and intercalated cells by differences in their enzyme content and further for revealing two subpopulations of intercalated cells. This method promises to provide a useful approach for studying the principal and intercalated cell populations in various metabolic states. PMID- 3039856 TI - Sample site selection for tracer studies applying a unidirectional circulatory approach. AB - The optimal arterial or venous sites for infusion and sampling during isotopic tracer studies have not been established. This study determined the relationship of plasma and tissue enrichment (E) when isotopes were infused in an artery and sampled from a vein (av mode) or infused in a vein and sampled from an artery (va mode). Adult dogs were given primed constant infusions of [3-13C]lactate, [1 13C]leucine, and 14C-labeled bicarbonate. Simultaneous samples were drawn from the vena cava, aortic arch, and breath. Tissue samples were removed from skeletal muscle, liver, kidney, and gut. Breath samples were analyzed for 14CO2 by liquid scintillation counting and plasma isotopic enrichments of [13C]lactate, [13C]leucine, and alpha-[13C]ketoisocaproate (KIC) were determined by gas chromatography-mass spectrometry. By using the va mode, the plasma E for lactate and leucine were 30-40% above tissue E. The av mode provided an accurate reflection of tissue E for lactate, which equilibrates rapidly with tissues, and a reasonable estimate for leucine, which exchanges more slowly. The isotopic enrichment of plasma KIC more directly reflected tissue leucine E than did plasma leucine E, and KIC enrichment was insensitive to sampling site. We also evaluated theoretically a circulatory model that predicts venous isotopic enrichments when the va mode is used. We conclude that the av mode is optimal but that the problems arising from use of the va mode can be overcome by use of a metabolic product (i.e., KIC) or by calculation of venous specific activity with our circulatory mode. PMID- 3039857 TI - Evaluation of a parathyroid hormone antagonist in an in vivo multiparameter bioassay. AB - The antagonist properties of a bovine parathyroid hormone analogue ([Tyr34]bPTH (7-34] amide were quantitatively assessed in vivo in a multiparameter assay to estimate the potency of the antagonist against the major actions of PTH. The analogue inhibited PTH-stimulated urinary excretion of phosphate and adenosine 3',5'-cyclic monophosphate in vitamin D-deficient thyroparathyroidectomized rats in a dose-dependent manner. At a molar dose ratio as low as 5:1 of antagonist to PTH, partial inhibition occurred. PTH stimulates the activity of 25 hydroxyvitamin D3-1 alpha-hydroxylase in renal proximal tubules. When coinfused with PTH, this analogue completely inhibited PTH-stimulated 1 alpha-hydroxylase activity at a molar dose ratio of 25:1 of antagonist to PTH and partially inhibited the activity at a molar dose ratio of 10:1. The analogue revealed no PTH-like agonist activity for stimulation of the 1 alpha-hydroxylase. Taken together, these studies indicate that [Tyr34]bPTH-(7-34) amide is a potent antagonist of several of the parameters of PTH action in vivo and demonstrate the feasibility of designing a PTH antagonist that can interact simultaneously with all the PTH receptors responsible for the hormone's major actions in vivo. PMID- 3039859 TI - Improved separation method for rat proximal and distal renal tubules. AB - We modified and improved enzyme digestion and density gradient separation procedures to obtain fractions of proximal and distal renal tubules with high yield and viability. Kidneys from two anesthetized adult Wistar rats were flushed with Krebs-Henseleit buffer (KHB) and then perfused in situ with recirculated KHB containing collagenase and hyaluronidase at 125 mmHg. Cortices were excised, minced, and incubated in KHB containing enzymes for 35 min at 37 degrees C. Dissociated tubules were removed at 10-min intervals, rinsed, and placed in KHB containing 10% calf serum, vitamins, and amino acids at 4 degrees C. Separation was achieved by suspending the tissue in 45% isosmotic Percoll layered over an undiluted Percoll cushion and centrifuging. Proximal tubules sedimented near the cushion. Distal segments were isolated in the uppermost bands of a second 35% Percoll separation. Viability was greater than 95% as measured by lactate dehydrogenase leakage and quantitated by oxygen consumption and ATP content. Basal oxygen consumption was greater than 33 nmol O2 X min-1 X mg protein-1 in all fractions and was stimulated by succinate and inhibited by amiloride and ouabain. Basal ATP content averaged 9.7 nmol/mg ATP. An average 3.3-fold separation for the proximal fraction and 24.5-fold separation for the distal fraction was assessed by the enrichment of six specific enzyme markers, with several of the markers indicating separations up to 32-fold. Isolated tubules also displayed functional responses to parathyroid hormone and vasopressin. Distal, but not proximal, segments demonstrated significantly increased adenosine 3',5'-cyclic monophosphate formation with vasopressin. PMID- 3039858 TI - EGF-induced mitogenesis in proximal tubular cells: potentiation by angiotensin II. AB - The mitogenic effect of epidermal growth factor (EGF) and the characteristics of EGF binding were studied on primary cultures of rabbit proximal tubular cells. EGF was found to be a potent mitogen and stimulated DNA synthesis 18-fold above the level observed in quiescent cells. Using 125I-EGF as a ligand, two classes of specific EGF receptors were identified on the proximal tubular cell in culture, i.e., a high-affinity receptor with a dissociation constant (Kd) of 0.3 nM and maximal binding (Bmax) of 1.7 X 10(4) receptors/cell and a low-affinity receptor with a Kd of 1.9 nM and Bmax of 5.3 X 10(4) receptors/cell. Because angiotensin II (ANG II) appeared to possess many properties common to growth factors, we also examined the interaction of ANG II and EGF on these cells. ANG II was not mitogenic, but it potentiated the mitogenic effect of EGF with a maximal effect at 10(-9) M. The dose-response curve of EGF-induced mitogenesis was shifted to the left in the presence of 10(-9) M ANG II, decreasing the approximate half maximal stimulatory concentration from 3 X 10(-8) to 5 X 10(-9) M. ANG II also stimulated prostaglandin E2 (PGE2) release, but inhibition of basal and ANG II stimulated PGE2 synthesis had no effect on mitogenesis. ANG II had no effect on the binding of EGF to the high-affinity receptor from 1 to 20 h and did not alter receptor downregulation. ANG II (10(-9) M) had no effect on cell protein content, RNA and protein synthesis, Na+-H+ antiport, and intracellular free Ca2+ concentration. Higher concentrations of ANG II (5 X 10(-8) to 5 X 10(-6) M) led to a rapid and transient dose-dependent rise in cytosolic free Ca2+ concentration. These studies demonstrate that ANG II potentiates EGF-induced mitogenesis at one or more postreceptor steps that may include small changes in cytosolic Ca2+ concentration. PMID- 3039860 TI - Dopaminergic stimulation of cAMP accumulation in cultured rat mesangial cells. AB - Dopamine (DA) alters renal hemodynamics, and DA receptors have been demonstrated in isolated glomeruli. To determine the glomerular cell type bearing DA receptors, we studied the effect of dopaminergic agonists and antagonists on adenosine 3',5'-cyclic monophosphate (cAMP) accumulation in rat glomerular mesangial and epithelial cells in culture. DA caused a marked dose- and time dependent increase in cAMP accumulation in mesangial but not epithelial cells. The stimulatory effect of DA was abolished by the DA antagonists haloperidol, trifluoperazine, and cis-thiothixene but not by beta- or alpha-adrenergic or histamine antagonists. Similar to the effects of DA, two dopamine type I receptor agonists, fenoldopam and SKF 38393, markedly stimulated cAMP accumulation in the mesangial cells. Moreover, the effects of DA were blocked by Sch 23390, a specific DA1 receptor antagonist, but not domperidone, a specific DA2 antagonist. These results show that DA regulates cAMP accumulation in mesangial cells via DA1 type receptors. PMID- 3039861 TI - Body composition, food intake, and brown fat thermogenesis in pregnant Djungarian hamsters. AB - Strategies for balancing energy storage and expenditure during pregnancy were examined in the Djungarian hamster. In expt 1, pregnant hamsters lost nearly 50% of their body lipid stores, even though they showed significant increases in body weight and food intake during the latter half of pregnancy. The increase in body weight was accounted for by the growth of the uteri, fetuses, and placentas. The water and fat-free dry contents of the maternal carcasses did not differ from those of the unmated controls. In expt 2, brown fat cytochrome-c oxidase activity (mitochondrial content) was significantly lower in the late- but not early pregnant females relative to the nonpregnant controls. Specific GDP-binding levels did not differ significantly among these groups. Thus an overall decrease in total thermogenic activity in brown fat would be expected during late pregnancy. The loss of carcass lipid, despite decreased brown fat thermogenesis and increased food intake, suggests that substantial increases in energy expenditure occur in pregnant females that are not related to heat production in brown fat. The present results are not consistent with traditional models of energy balance during pregnancy. Some of the inconsistencies may be related to differences between hoarding and nonhoarding families of rodents, whereas others may be due to the fact that the traditional model is based on a possibly exceptional species, the laboratory rat. PMID- 3039862 TI - Diuretic and natriuretic properties of prestegane B, a mammalian lignan. AB - The effect of increasing doses of prestegane B, a synthetic lignan, was examined in the anesthetized normal rat, using clearance methodology. Increasing doses of prestegane B 0.5, 1.0, 2.0, and 5.0 mg) were administered intravenously in our separate groups of hydropenic rats. Urine flow increased by 2.8 +/- 0.3, 4.5 +/- 0.5, 7.7 +/- 0.5, and 18.2 +/- 0.8 microliters/min, respectively, above control values. The rise of urinary sodium secretion was of similar magnitude and averaged 0.4 +/- 0.1, 0.8 +/- 0.2, 1.1 +/- 0.3, and 2.4 +/- 0.3 mu eq/min, respectively. No significant change in urinary phosphate excretion was obtained in all groups of rats, and glomerular filtration rate remained constant from control to experimental clearance periods. The natriuretic effect of prestegane B observed in this in vivo model could be related to the inhibition of the Na+-K+ adenosine triphosphate activity demonstrated in vitro in previous studies from our laboratory. The action of this substance is likely to be situated beyond the proximal tubule, since urinary phosphate was not altered. Prestegane B mimics the effects of other endogenous diuretic and natriuretic hormones, but its site of action and its effect on renal hemodynamics are obviously different. PMID- 3039864 TI - [Epstein-Barr virus and rhinopharyngeal cancer]. PMID- 3039863 TI - Effect of upper respiratory infection on hearing in the ferret model. AB - In an effort to develop an adult animal model for acquired viral-induced hearing loss, three groups of mature ferrets were inoculated intranasally with respiratory viruses (influenza A/Port Chalmers, influenza B/Mass, and parainfluenza I), which have been implicated as causative agents in idiopathic sudden hearing loss (ISHL). All ferrets challenged with influenza A/Port Chalmers (A/PC) exhibited clinical signs of infection, but neither of the other two groups exhibited such signs. Conductive and/or sensorineural hearing losses were demonstrated in eight of 15 ferrets challenged with influenza A/PC and four of 10 ferrets in the group challenged with influenza B/Mass by brainstem auditory evoked responses (BAERs) and acoustic immittance. None of the four ferrets infected with parainfluenza I demonstrated auditory dysfunction. When auditory changes were observed, nasal washes were performed, and tissues of the middle and inner ears were collected for viral assay and morphologic examination. All ferrets demonstrating auditory changes were shown to be infected by isolation of virus from nasal tissues. Virus was isolated from eustachian tube tissue in six of the eight ferrets infected with influenza A/PC, which also demonstrated altered BAERs and one of the four infected with influenza B/Mass, which also had auditory changes. Virus was isolated from the middle ear of two ferrets infected with influenza A/PC and from the inner ear of another two ferrets from the same group. These data suggest that influenza A/PC has a greater effect on auditory function in the ferret model than either influenza B/Mass or parainfluenza I and that the ferret may be an appropriate adult model to examine the etiologic role of viral upper respiratory tract infections in some acquired hearing impairments. PMID- 3039865 TI - Variations on the "dilution" method for reconstituting cytochrome c oxidase into membrane vesicles. AB - A method for the rapid incorporation of cytochrome c oxidase into membranes has been developed. This method essentially consists of obtaining a preparation of the enzyme in which it is isolated and then dissolving it in a medium containing 0.5% of the detergent Tween 20, which gives a final concentration of 0.0125% after reconstitution. These studies revealed an optimal ratio of 1 microgram of enzyme to 5 mg of phospholipids. A similar optimal ratio was found when the amount of protein was varied. The optimum temperature was found to be 30 degrees C. Without a peak value being reached, it was found that the best reconstitution was obtained at pH 7.0-8.0. When measurements were performed either with a fluorescent cyanine (DiSC3) or by the uptake of tetraphenylphosphonium, it was found that the enzyme, with cytochrome c added to the outside, was capable of generating a membrane potential that was negative inside. Using the same procedure, the enzyme could also be reconstituted into vesicles of yeast plasma membrane. The procedure, then, seems adequate for incorporating cytochrome c oxidase into different kinds of membrane vesicles. PMID- 3039866 TI - Hybridization as a technique for studying interchain interactions in the catalytic trimers of aspartate transcarbamoylase. AB - Since subunit interactions in regulatory enzymes mediate the ligand-promoted conformational changes responsible for their allosteric properties, it is necessary to have techniques for determining the effects of ligands and mutational alterations on the strength of the interchain interactions. In aspartate transcarbamoylase from Escherichia coli, the multiple interchain interactions are so linked that it is difficult to study them separately. Therefore, we have focused on the nonallosteric catalytic trimers isolated from the holoenzyme and have used the rate of hybrid formation between native and succinylated protein as a measure of the dissociation of the trimers into single polypeptide chains. Although catalytic trimers exhibit no evident dissociation in sedimentation studies at 10(-8) M, incubation of mixtures of native and succinylated trimers for long periods of time (days) yielded hybrids which are readily detected by polyacrylamide gel electrophoresis. This sensitive technique was used to demonstrate that the substrate, carbamoylphosphate, and the bisubstrate analog, N-(phosphonacetyl)-L-aspartate, cause a marked strengthening of the interchain interactions, whereas the inhibitor, sodium pyrophosphate, at concentrations as low as 10 mM, promotes dissociation of the trimers. This weakening of the interchain interactions by pyrophosphate facilitated the isolation and purification of functionally competent hybrid trimers by a technique which was much more convenient and provided higher yields than previous, more drastic methods which employed urea or guanidine hydrochloride to cause dissociation of the trimers. The hybridization technique was useful in studying the effects of mutational alterations on the strength of the interchain interactions and the ability of active and inactive mutants to bind pyrophosphate. PMID- 3039867 TI - Flameless atomic absorption spectrophotometric determination of platinum in tissues solubilized in hyamine hydroxide. AB - Processing biological samples by solubilization in Hyamine hydroxide (methylbenzethonium hydroxide) as an alternative to wet ashing with concentrated nitric acid for atomic absorption determination of platinum has been explored. The concentrations of platinum in tissues removed from rats 2 h after they had received the antitumor drug Carboplatin (60 mg/kg, iv) were comparable using either of the procedures, indicating the utility of tissue solubilization. Limits of detection were also comparable between the Hyamine hydroxide (0.25 microgram platinum/g) and nitric acid (0.15 microgram/g) procedures. The solubilization method, however, has advantages over wet digestion because of its simplicity and greater safety. PMID- 3039868 TI - Rapid alkaline blot-transfer of viral dsRNAs. AB - The double-stranded genomic RNAs of reovirus and bluetongue virus can be transferred very efficiently from either sodium dodecyl sulfate-polyacrylamide gels or NuSieve agarose gels onto several nylon membranes. After a brief acid depurination treatment, viral dsRNAs from the gels are transferred at room temperature using 0.2 N NaOH as the transfer medium. Four blots can be obtained within 1 h and each blot contains 15-20% of the input RNA sample. These blots can be used immediately without baking in vacuo. Less than 5% of the "fixed" dsRNAs are removed after repeated washings of the membrane blots. As little as 10 pg of the genomic dsRNA segment can be detected in this alkaline Northern blot. A 20- to 50-fold increase in resolution and sensitivity over traditional Northern blots is routinely achieved. These alkaline blots can be reused 6-10 times after appropriate strip washing and proper handling. PMID- 3039869 TI - The use of biotinylated monoclonal antibodies and streptavidin affinity chromatography to isolate herpesvirus hydrophobic proteins or glycoproteins. AB - A streptavidin/biotin-based immunoaffinity system was optimized to isolate herpesvirus (human cytomegalovirus) immediate early proteins or late glycoproteins from crude infected cell lysates. Biotinylation of the primary antibody by biotin substitution of epsilon amino groups was superior to biotin substitution of sugar residues. Biotinylation of the primary antibody was superior to that of a secondary antibody. A biotin substitution of approximately 8 M biotin/M antibody allowed for maximal recovery of viral antigens. The streptavidin/biotin-based immunoaffinity system can allow for relatively pure preparations of viral antigens that may be used for functional, immunological, or structural studies. PMID- 3039870 TI - In vitro sodium bisulfite mutagenesis of restriction endonuclease recognition sites. AB - Sodium bisulfite treatment of single-stranded DNA deaminates exposed cytosine residues to form uracil, resulting in cytosine-to-thymidine transition mutations following DNA replication. We have used this reaction in vitro to destroy the recognition sequences for the restriction endonucleases HindIII and XmaI in the aminoglycoside 3'-phosphotransferase I coding region of plasmid pUC4K. This procedure should be applicable to the mutation of any recognition sequence of restriction endonucleases which generate cytosine-containing single-stranded ends. The possibility of mutagenesis of restriction sites to generate stop codons in coding regions is discussed. PMID- 3039871 TI - Liquid chromatographic assay of beta-lactamase inhibitors in human serum and urine using a hollow-fiber postcolumn reactor. PMID- 3039872 TI - Effect of temperature on the kinetics of free radical formation during the pyrolysis of some amino acids. PMID- 3039873 TI - Effect of intrathecal bupivacaine on somatosensory evoked potentials following dermatomal stimulation. AB - The effect of spinal anesthesia with 3.6 +/- 0.1 ml (mean +/- SEM) of 0.5% bupivacaine on early (less than 150 msec) somatosensory evoked potentials (SEPs) with electrical stimulation of the L1 and S1 dermatomes was examined in 12 patients. The mean level of sensory analgesia (pinprick) was T8,9 +/- 1.0 (+/- SEM) and the mean degree of motor blockade was 1.3 +/- 0.1 (Bromage scale). Intrathecal bupivacaine significantly (P less than 0.05) decreased the amplitude of all SEP components after stimulation of the L1 dermatome and most components during stimulation of the S1 dermatome. Intrathecal bupivacaine also increased the latency of SEPs (P less than 0.05) of both dermatomes. The L1 SEP disappeared in 7 and the S1 SEPs in 5 of the 12 patients during neural blockade. In three patients the SEPs disappeared at both locations. Sensory thresholds increased significantly during blockade. We found no correlation between decrease of amplitude and degree of motor blockade or level of sensory analgesia. Thus, intrathecal plain bupivacaine has a strong depressant effect on the neural afferent transmission as assessed by SEPs. However, despite clinically effective blockade as assessed by pinprick and motor blockade nerve potentials after nociceptive stimulation within the area of sensory block were often able to pass to the cerebral cortex. PMID- 3039874 TI - Radiation sterilization of surgical instruments with a consideration of metal shielding on sterilization efficiency. AB - The feasibility of the use of radiation for sterilization of surgical instruments was evaluated. Two aspects were considered: radiation biology of relevant microorganisms, that is, bacterial spores and viruses, and shielding and radiation protection by the metal of the instruments. After proper cleaning and hot water machine washing, surgical instruments carry few, if any contaminants; however, subsequent handling increases the contamination load. Although large instruments may attenuate as much as 30% of the incident radiation, spores dried on the metal are sensitized to irradiation by some 40%. A dose of 25 kGy (2.5 Mrad) is adequate to inactivate a potential contamination load of approximately 10(7) bacterial spores or approximately 10(4) viruses. Therefore, 25 kGy will provide a high sterility assurance level, and can be recommended with a considerable degree of confidence for hospital-based sterilization of surgical instruments. PMID- 3039875 TI - Cytomegalovirus: epidemiology and infection control. AB - Cytomegalovirus (CMV) causes asymptomatic infection in most individuals but can produce devastating illness in immunocompromised hosts and in a small proportion of congenitally infected babies. New techniques in molecular biology have provided insights into the epidemiology and transmission of CMV. Children in day care, their parents, and sexually active individuals, especially homosexual men, are now known to be at particular risk for acquiring CMV. Recent studies show that the risk of CMV acquisition by health care workers is similar to the risk to the general public. Health care workers should be aware of the wide range of clinical manifestations, methods of laboratory diagnosis, and current limitations of treatment of CMV. Careful handwashing and avoidance of excretions and secretions are recommended to decrease the transmission of CMV in the hospital. PMID- 3039876 TI - [Protein C, protein S]. AB - Protein C is a potent inhibitor of blood coagulation, and, in addition, appears to be a profibrinolytic agent. In a first step, protein C must be converted to a serine protease. This activation is catalyzed by a complex formed between thrombin and thrombomodulin, an endothelial cell surface protein. Activated protein C exhibits its anticoagulant activity through the proteolytic inactivation of two blood coagulation cofactors, factors Va and VIIIa. This reaction requires phospholipids, originating from platelets or endothelial cells, and a cofactor protein, protein S. Protein S enhances the binding of activated protein C to phospholipids. In addition, activated protein C stimulates fibrinolysis, through the inactivation of the tissue plasminogen activator (tPA) inhibitor. An isolated constitutional, quantitative or qualitative, protein C or protein S deficiency increases the risk of thrombosis, the clinical features are different in the rare cases of homozygous protein C deficiency (neonatal purpura fulminans) or in the heterozygous patients (recurrent venous thrombosis in young adults). Acquired deficiency in protein C and S had been observed in liver disease, during vitamin K antagonists or L-Asparaginase treatment, and in disseminated intravascular coagulation. PMID- 3039877 TI - [Different localization of thrombomodulin]. AB - Thrombomodulin (TM) is the endothelial cofactor of the anticoagulant protein C system. The distribution of TM in the organism was studied in the rabbit using a goat anti TM, affinity-purified antibody and a peroxidase-labelled antigoat immunoglobulin. TM antigen was found on the endothelial surface of all blood vessels: capillaries, arteries and veins. The reaction was specific: connective tissue, smooth and striated muscle bone, cartilage, nerve tissue, secretary epithelia and all parenchyma studied were not strained. Moreover TM antigen was present on the surface of serosa: peritoneum, pericardium and pleura as well as on synovial membranes and on arachnoid, all along the central nervous system. It was absent from pia and dura mater. The antigen was found only after formalin fixation on the vessels and body cavities surface on which it lies. This observation shows that the antigen is easily detached from these surfaces and suggest a possible mobility of this endothelial molecule for which it lies. This observation shows that the antigen is easily detached from these surfaces and suggest a possible mobility of this endothelial molecule for which production and function sites might differ. PMID- 3039878 TI - Receptor-mediated O2- release by alveolar macrophages and peripheral blood monocytes from smokers and nonsmokers. Priming and triggering effects of monomeric IgG, concanavalin A, N-formyl-methionyl-leucyl-phenylalanine, phorbol myristate acetate, and cytochalasin D. AB - Receptor-mediated superoxide (O2-) release by alveolar macrophages and peripheral blood monocytes from smokers and nonsmokers was studied in vitro. When the cells were incubated with monomeric IgG or monomeric Fc(IgG) fragment, no cell O2- release was observed. However, when cytochalasin D (Cyto D) was subsequently added to the cell suspension, we observed a markedly enhanced O2- release. Neither Cyto D alone nor the double stimulation of following Cyto D with monomeric IgG induced O2- release. Concanavalin A (Con A) also had a priming effect on O2 release in combination with Cyto D, as did monomeric IgG or monomeric Fc(IgG) fragment. On the other hand, heat-aggregated IgG, N-formyl methionyl-leucyl-phenylalanine (FMLP), or phorbol myristate acetate (PMA) induced O2- release without the addition of Cyto D. Thus, we observed 2 different mechanisms in the receptor-mediated O2- release by alveolar macrophages and peripheral blood monocytes. Alveolar macrophages from smokers, which had a higher affinity and a larger number of monomeric IgG binding sites per cell than those from nonsmokers, were more reactive to the double stimulation of following monomeric IgG with Cyto D than to that of Con A and Cyto D, FMLP, or PMA, but for peripheral blood monocytes it was the reverse. We conclude that the binding of monomeric IgG to the Fc(IgG) receptor of alveolar macrophages or peripheral blood monocytes results in a priming effect on the cells for O2- release, and that the regulation of receptor-mediated O2- release by alveolar macrophages differs at least in part from that of peripheral blood monocytes. PMID- 3039879 TI - Activity of rifabutin alone or in combination with clofazimine or ethambutol or both against acute and chronic experimental Mycobacterium intracellulare infections. AB - Rifabutin (ansamycin LM 427) when given alone in doses of 5, 10, or 20 mg/kg showed limited in vivo activity against acute or chronic Mycobacterium intracellulare infections studied in beige C57B1/6, or S/W mice. Prophylactic studies in S/W mice in which chemotherapy was given for 3 wk prior to challenge showed slight improvement. Combination chemotherapy consisting of rifabutin (10 mg/kg) and clofazimine (20 mg/kg) achieved complete sterilization of M. intracellulare infections in spleen and lung when initiated immediately after challenge. Inclusion of ethambutol did not offer additional benefit. If chemotherapy is initiated 3 wk after challenge (established infections), the activity of this double drug combination is less effective. PMID- 3039880 TI - Effect of rifabutin on the phagocytosis and intracellular growth of Mycobacterium intracellulare in mouse resident and activated peritoneal and alveolar macrophages. AB - In order to understand the discrepant results between in vitro and in vivo activity of rifabutin (LM 427) against Mycobacterium intracellulare, we studied its effects upon the phagocytosis of M. intracellulare by resident and activated mouse peritoneal and alveolar macrophages and upon growth of M. intracellulare inside these cells. Rifabutin (ansamycin, LM 427) had no effect on the viability of resident or activated peritoneal or alveolar macrophages or on the phagocytosis of M. intracellulare by resident peritoneal macrophages. In activated peritoneal and alveolar macrophages, it had a dose-related effect on the phagocytic index and intracellular growth of M. intracellulare. Macrophages from mice pretreated with 10 or 20 mg/kg of rifabutin either orally or intraperitoneally for 4 to 10 days showed inhibition of intracellular growth of M. intracellulare in a dose-related manner. The route of administration or further addition of the drug did not enhance the inhibitory activity of these macrophages. These results indicate that rifabutin has the potential of killing M. intracellulare persisters inside macrophages. PMID- 3039881 TI - Behavior and clinical relevance of histamine and leukotrienes C4 and B4 in grass pollen-induced rhinitis. AB - The release kinetics of histamine and leukotrienes C4 (LTC4) and B4 (LTB4) were investigated in nasal secretions of 10 patients with hay fever after antigen challenge. High levels of biologically active histamine were found in nasal washes from asymptomatic allergic and normal subjects. With repeated lavages, the amount of histamine recovered dropped markedly. Grass pollen challenge was followed by a significant (p less than 0.05) dose-dependent and time-limited (5 min) increase in histamine level in 7 of 10 patients; these values, however, were lower than those found in basal conditions. In 8 of 10 patients with hay fever, antigen challenge induced a significant (p less than 0.05) dose-dependent increase in LTC4 level, which persisted for 30 min. The LTC4 generation was well correlated with the appearance of allergic symptoms; LTB4 production was found in 2 patients only. A different pattern of symptoms was observed after in vivo nasal stimulation with histamine and LTC4. Histamine caused sneezing, itching, rhinorrhea, and nasal obstruction; conversely, the main symptom induced by LTC4 was a more pronounced and longer lasting nasal obstruction. PMID- 3039882 TI - Prevention of lower respiratory herpes simplex virus infection with acyclovir in patients with the adult respiratory distress syndrome. AB - Herpes simplex virus (HSV) type I commonly occurs in the lower respiratory tract (LRT) of seriously ill patients, particularly those with the adult respiratory distress syndrome (ARDS), but it is not known whether HSV is a benign mucosal colonizer or a pathogen. The aims of this study were to determine whether the antiviral agent acyclovir could prevent this occurrence, and if so, whether prevention improved the outcome. Forty-five patients with ARDS underwent double blind randomization into a treatment group (22 subjects) who received prophylactic acyclovir intravenously, 5 mg/kg every 8 h, and a control group (23 subjects). Upper and lower respiratory secretions were examined for the presence of HSV before randomization and twice weekly thereafter. Seven patients were excluded because of HSV detection prior to treatment. There were no significant differences between the remaining 17 acyclovir and 21 control patients in age, sex, distribution of primary diagnostic categories, and severity of primary illness. Only 1 patient (6%) in the acyclovir group developed HSV after treatment compared with 15 (71%) in the control group (p less than 0.001), but there was no improvement in the acyclovir group in the severity of respiratory failure, the duration of ventilator support (acyclovir, 20 +/- 19 days; control, 14 +/- 11 days), or mortality (acyclovir, 8 of 17, 47%; control, 9 of 21, 43%). We conclude that acyclovir is effective in preventing the high incidence of HSV in patients with ARDS, but that this prevention does not improve outcome. Routine prophylaxis of HSV is not recommended. PMID- 3039883 TI - The role of arachidonate mediators in peroxide-induced lung injury. PMID- 3039884 TI - Lipid peroxides and the coronary circulation. PMID- 3039885 TI - Cytochrome P-450-related arachidonic acid metabolites. AB - Arachidonic acid (AA) can be metabolized to diverse products that differ widely in their biologic activities. Depending on the cell type, AA can be metabolized by 3 pathways: cyclooxygenase, lipoxygenases, and cytochrome P-450 monooxygenases. Disease states, injury, and stress, will influence the enzymes that regulate the breakdown of AA and may favor the generation of products not usually associated with a tissue. Studies on renal eicosanoid mechanisms (all AA metabolites) can serve as a paradigm for other tissues and organs. Discrete stimulation of renal AA metabolism can occur-localized to specific cells within segments of the nephron; for example, cells of the medullary segment of the thick ascending limb of the loop of Henle (mTALH) have high cytochrome P-450 monooxygenase activity, through which novel AA metabolites are formed. These metabolites have characteristic biologic activities, inhibition of Na+-K+-ATPase activity and relaxation of blood vessels, and can be stimulated by peptide hormones. Cytochrome P-450-dependent monooxygenases are also present in the lung where, in their suggested capacity as oxygen sensors, they generate metabolites of AA that modify pulmonary vasomotion. PMID- 3039886 TI - Failure of Centers for Disease Control criteria to identify hepatitis B infection in a large municipal obstetrical population. AB - The Centers for Disease Control (CDC) has recommended screening pregnant women from high-risk populations for hepatitis B surface antigen (HBsAg). To assess the adequacy of the risk criteria, all women presenting for delivery to a large municipal hospital were screened. Sera from 5356 women were tested, and questionnaires designed to identify women at high risk were completed by 78% of these patients. Sixty-four women were found to be HBsAg seropositive (1.2%). If the CDC criteria had been applied for screening, 30 of the seropositive mothers (47%) would not have been identified. Women from some Latin American and Caribbean countries not recommended for screening were found to have a relatively high prevalence of hepatitis B infection. Reluctance to give a history of venereal disease or illicit drug use may be another factor in the failure of the CDC screening strategy. To achieve effective immunoprophylaxis of newborns, all pregnant women should be screened for HBsAg carriage. PMID- 3039887 TI - Resolution of oral hairy leukoplakia during therapy with 9-(1,3-dihydroxy-2 propoxymethyl)guanine (DHPG). PMID- 3039888 TI - Cytomegalovirus-associated tubulointerstitial nephritis in an allogeneic bone marrow transplant recipient. PMID- 3039889 TI - Update on hepatitis B prevention. Recommendations of the Immunization Practices Advisory Committee. Centers for Disease Control, Department of Health and Human Services. AB - Since plasma-derived hepatitis B vaccine was licensed for use in the United States in 1982, use of the vaccine in high-risk health care professions has been modest, despite widespread participation in the establishment of vaccination programs among these institutions. The incidence of hepatitis B virus infection has continued to increase in the past 5 years, presumably because vaccination programs have failed to reach the major risk groups of homosexual men, parenteral drug abusers, and heterosexually active persons with multiple sexual partners. Increased efforts toward the development of programs to vaccinate persons in all high-risk groups are recommended. The immunogenicity, safety, efficacy, and precautions involved in the use of a new recombinant DNA vaccine (licensed in 1986) are also discussed, and the need for booster doses of plasma-derived vaccine are evaluated. PMID- 3039890 TI - Neurologic manifestations of infection with human immunodeficiency virus. Clinical features and pathogenesis. AB - Subacute encephalitis caused by infection of the central nervous system by the human immunodeficiency virus (HIV) is the most frequent cause of neurologic dysfunction in patients with the acquired immunodeficiency syndrome (AIDS). This disorder results in progressive cognitive, motor, and behavioral abnormalities in at least two thirds of patients with AIDS. Pathologic evidence of subacute encephalitis is found in 90% of these patients at autopsy. Human immunodeficiency virus is also the etiologic agent of aseptic meningitis, a disease that can occur at the time of seroconversion. Other neurologic disorders frequently associated with HIV include peripheral neuropathies and vacuolar myelopathy. Thus, HIV is neurotropic and may enter the central nervous system early in the course of infection. Neurologic disease may be the only clinical manifestation of HIV infection. Although mechanisms of pathogenesis are unclear, cells of monocyte macrophage lineage may be important in viral spread to and within the central nervous system. Effective antiviral therapy will probably require penetration of drugs across the blood-brain barrier. PMID- 3039891 TI - Ascorbic acid and the eye with special reference to the lens. PMID- 3039892 TI - Ascorbic acid and cataract. PMID- 3039893 TI - Ascorbic acid and cancer. PMID- 3039894 TI - Recovery and probable persistence of cytomegalovirus in human inner ear fluid without cochlear damage. AB - Cytomegalovirus (CMV) was recovered from a 5-month-old infant with probable congenital infection. In life, no hearing impairment had been observed. Auditory brain stem evoked responses were bilaterally intact. At necropsy, both temporal bones were morphologically normal, as demonstrated by light and electron microscopy. Sensory hair cells of the organ of Corti appeared intact. Cytomegalovirus was recovered from a mixture of perilymph and endolymph, but not the brain, CSF, or vitreous humor. This appears to be the first report of an individual with an inner ear CMV infection in which neither structural nor functional alterations of the inner ear were apparent. This case also suggests that CMV can persist within the inner ear for prolonged periods following congenital infection. PMID- 3039895 TI - [Alkaloids from the seeds of Sarcopharyngia crassa (Benth.) Boiteau et Allorge]. PMID- 3039896 TI - [Assay of caffeine, theobromine, catechin and epicatechin by high performance liquid chromatography in an extract of fresh stabilized cola seeds]. PMID- 3039897 TI - [Experimental study of the toxicity of arils from Blighia sapida (Sapindaceae) in relation to the poisoning of children of Katiola (Ivory Coast)]. PMID- 3039898 TI - [Hepatic arterial lipiodolization and x-ray computed tomography diagnosis of malignant liver tumors. Experience apropos of 33 cases]. PMID- 3039899 TI - The pestiviruses: where do they belong? AB - In this introduction to the meeting, the author specifies the place of pestiviruses among RNA viruses and briefly summarizes the main characteristics from this genus. PMID- 3039900 TI - Molecular cloning of the bovine viral diarrhea virus genomic RNA. AB - The genomic RNA from Bovine Viral Diarrhea Virus (BVD) was cloned in E. coli. The complete sequence has been obtained and the genomic organization has been deduced. Some of the BVD specific cDNA have been expressed in bacteria, eucaryotic cells and in vitro. We suggest that BVDV belongs to a new family different from Togaviridae and Flaviviridae. PMID- 3039901 TI - Pathogenesis and epidemiology of border disease. AB - The pathogenesis of BD virus infection of sheep is reviewed briefly. The most serious consequences occur when susceptible pregnant sheep are infected. The virus crosses the placenta readily and can cause foetal death with resorption, mummification or stillbirths. Other lambs survive, however, and are born with varying degrees of tremor and/or hairy fleeces. Many such "hairy-shaker" lambs die shortly after birth but survivors gradually recover. These survivors plus other apparently normal lambs can be persistently infected with virus and excrete it constantly often for the rest of their lives. It is these persistently infected sheep that are the key to the epidemiology of the virus and may be responsible for its introduction into a susceptible flock. In addition, cattle grazed with sheep are potentially an important source of pestiviruses capable of causing BD since in some herds the prevalence of cattle persistently infected with pestivirus is one to three per cent. Results are presented of studies on antigenic relation ships among Scottish isolates of cattle pestiviruses (BVD) and BD viruses, and the conclusion drawn that while a single strain of BVDV may be suitable for use as a cattle vaccine this would not be very efficacious in preventing BD since two antigenically distinguishable strains of BD virus have been identified. An effective vaccine against BD would have to protect sheep against both these strains and ideally a cattle vaccine should also contain both strains. PMID- 3039902 TI - [9 case reports of insulinoma. Proposals for a current therapeutic strategy]. PMID- 3039903 TI - [Bronchial carcinoid tumor and Cushing's syndrome. Apropos of 3 cases]. PMID- 3039905 TI - [Hereditary nature of nephroblastoma. Apropos of a case of 3 siblings]. AB - Three nephroblastomas, including one bilateral tumour, have been detected in three children belonging to the same family and born from a consanguinous marriage. The clinical and diagnostic particularities compared to non familial "sporadic" nephroblastomas are studied. The pathogenesis with the incidence of chromosomal anomalies, the diagnosis and the frequency of bilateral diseases along with the associated deformities and a non invasive therapeutic attitude are discussed. PMID- 3039904 TI - [Malignant fibrohistiocytoma of bone. Apropos of 5 cases. Clinical and therapeutic aspects]. PMID- 3039906 TI - [Bilateral neuroblastoma. Study of 5 cases and review of the literature]. AB - Five cases of bilateral nephroblastoma out of total of 99 cases have been observed during a period of 18 years at the Children's Hospital of Tunis (5%). The frequency of associated anomalies, the familial incidence, the young age of the patients are underlined. The main therapeutic modalities and prognosis are envisaged. PMID- 3039907 TI - The molecular biology of the hepatitis B viruses. PMID- 3039908 TI - Cyclic AMP and other signals controlling cell development and differentiation in Dictyostelium. PMID- 3039909 TI - Receptors for epidermal growth factor and other polypeptide mitogens. PMID- 3039910 TI - Inhibition by ganciclovir of cell growth and DNA synthesis of cells biochemically transformed with herpesvirus genetic information. AB - The ability of LM cells, thymidine kinase-deficient LM cells (LMTK-), and LMTK- cells transformed to the LMTK+ phenotype by herpes simplex virus type 1 genetic information (LH7 cells) to anabolize the acyclovir congener ganciclovir was examined. About 50-fold more ganciclovir triphosphate was produced by LH7 cells than by either LM or LMTK- cells. Growth inhibition studies indicated that 180 and 120 microM ganciclovir were required to achieve 50% growth inhibition of LM and LMTK- cells, respectively; only 0.07 microM ganciclovir was necessary to achieve 50% inhibition of LH7 cells. DNA synthesis in the transformed cells was significantly reduced by ganciclovir treatment, whereas ganciclovir had little effect on DNA synthesis in the nontransformed cells. Alkaline sucrose gradient sedimentation analysis of transformed cellular DNA indicated that LH7 DNA synthesized in the presence of ganciclovir chased into mature DNA. Both LM and LH7 DNA synthesized in the presence of ganciclovir exhibited a concentration dependent reduction in the rate of elongation into mature DNA. Finally, [14C]ganciclovir was incorporated internally into the growing chains of LH7 cells. PMID- 3039912 TI - Rapid herpes simplex virus susceptibility testing using an enzyme-linked immunosorbent assay performed in situ on fixed virus-infected monolayers. AB - An enzyme-linked immunosorbent assay was performed directly on fixed, virus infected cell monolayers to rapidly determine the susceptibility of herpes simplex virus to antiviral drugs. Susceptibility determinations by this method correlated well with those obtained by plaque reduction assay and avoided some of the drawbacks of other methods currently available. PMID- 3039911 TI - Inhibitory effect of 2',3'-didehydro-2',3'-dideoxynucleosides on infectivity, cytopathic effects, and replication of human immunodeficiency virus. AB - It is generally accepted that human immunodeficiency virus (HIV) is the etiologic agent of the acquired immunodeficiency syndrome and related diseases. In this report, we demonstrate the antiviral effect of nucleoside analogs 2',3'-didehydro 2',3'-dideoxythymidine (DHT) and 2',3'-didehydro-2',3'-dideoxycytidine (DHC) by using human T-cell lymphotropic virus type I-carrying MT-4 cells, which are extremely susceptible to HIV infection. These agents efficiently inhibited the cytopathic effects and expression of HIV-specific antigens in MT-4 cells after infection of the virus. Both DHT and DHC also strongly blocked viral replication as determined by our quantitative bioassay system using a plaque-forming assay. These antiviral effects were obtained at concentrations at which the drugs produced little or no toxicity and were comparable to those with 3'-azido-3' deoxythymidine and 2',3'-dideoxynucleosides. These findings warrant further investigation of the use of DHT and DHC for the treatment of the acquired immunodeficiency syndrome and related diseases. PMID- 3039913 TI - Effect of epidural spinal cord stimulation on the activity of lateral vestibular nucleus neurons in the cat. AB - The activity of neurons in Deiters' lateral vestibular nucleus was recorded in decerebrate cats before, during and after spinal cord stimulation. An almost equal number of units were inhibited and excited early during stimulation. Later during stimulation the majority of units was inhibited. Early after cessation of stimulation an ever larger number of units were inhibited to an even larger extent (for about 2 imp/s on the average). Later after stimulus cessation the predominant inhibitory effect could still be noted, as well as excitation in some units. The results could support the hypothesis that the inhibition of Deiters' neurons during and for some time after epidural cord stimulation may play a part in the decrease of limb spasticity. The mechanism of inhibitory and excitatory unitary responses, side effects during stimulation and differences between the experimental model and human state are discussed. PMID- 3039914 TI - Cytochrome P-450 cholesterol 7 alpha-hydroxylase: inhibition of enzyme deactivation by structurally diverse calmodulin antagonists and phosphatase inhibitors. AB - Cytochrome P-450 cholesterol 7 alpha-hydroxylase (P-450Ch7 alpha) catalyzes the first and rate-limiting step in the conversion of cholesterol to bile acids. Incubation of rat liver microsomes in 4-(2-hydroxyethyl)-1 piperazineethanesulfonic acid buffer resulted in a time-dependent deactivation of P-450Ch7 alpha which was markedly accelerated by the nonionic detergent Tween 80. Microsomal NADPH-cytochrome P-450 reductase and cytochrome P-450-dependent 7 ethoxycoumarin O-deethylase activities were unaffected under these conditions, evidencing the selectivity of the deactivation process for P-450Ch7 alpha. The rate (t 1/2 = 15-19 min at 37 degrees C) and maximal extent of P-450Ch7 alpha deactivation (greater than or equal to 90%) were both unaffected by the presence of cytosolic proteins and were also not dependent on the initial enzyme level, as shown using liver microsomes isolated from untreated, cholestyramine-fed, and xenobiotic-induced rats exhibiting an eight-fold range in P-450Ch7 alpha activity. Scavengers for reduced oxygen species were also without effect. P 450Ch7 alpha was stabilized some six- to sevenfold (t 1/2 = 94-143 min) by the phosphatase inhibitor NaF. Of a series of other phosphatase inhibitors examined, including, among others, EDTA, vanadate, and molybdate, only phosphate-containing compounds and the calmodulin antagonist trifluoperazine, and inhibitor of the Ca2+-calmodulin-dependent phosphatase calcineurin, effectively stabilized P 450Ch7 alpha. Modulation of P-450Ch7 alpha deactivation by these inhibitors generally paralleled their effects on isolated calcineurin. A variety of structurally diverse calmodulin antagonists examined were also found to effectively protect P-450Ch7 alpha from deactivation; these include calmidazolium and tamoxifen (IC50 = 25 to 50 microM), chlorpromazine, thioridazine, amitriptyline, imipramine, and the naphthalene sulfonamide compound W-7 (IC50 = 50 to 300 microM). Structure-activity analysis of several phenothiazines and their derivatives indicated that although little activity was exhibited by the sulfoxides, some protection was provided by the corresponding sulfones. On the basis of these observations, various models for the molecular basis of enzyme deactivation are considered, including the hypothesis that a calcineurin-like microsomal phosphatase mediates deactivation of this cytochrome P-450 enzyme. PMID- 3039915 TI - Modulation of nitrate uptake in Anacystis nidulans by the balance between ammonium assimilation and CO2 fixation. AB - Gradual inhibition of ammonium assimilation in Anacystis nidulans cells by increasing concentrations of 5-hydroxylysine resulted in a progressive enhancement of nitrate uptake. For 5-hydroxylysine-treated cells, the magnitude of the inhibition of nitrate uptake promoted by added ammonium was dependent on the ammonium assimilation capacity. In cells with a moderate ammonium assimilation activity, acceleration of CO2 fixation induced by bicarbonate addition antagonized the negative effect of ammonium, allowing full nitrate uptake activity. The results support the contention that nitrate utilization is under the feed-back control exerted by products of its own assimilation via ammonium, the inhibitory effect being potentiated by ammonium addition and alleviated by enhanced CO2 fixation. Results of amino acid analysis in cells exhibiting different capacities to utilize nitrate speak against these compounds as direct effectors of nitrate uptake. PMID- 3039916 TI - Modification of the properties of S2 state in photosynthetic O2-evolving center by replacement of chloride with other anions. AB - Properties of the S2 state formed in photosystem II membranes in which Cl- had been replaced by various anions were investigated by means of thermoluminescence measurements and low temperature EPR spectroscopy. The Br--substituted membranes showed the normal thermoluminescence B-band arising from S2Q-B charge recombination, whereas the SO2-4-, F--, CH3COO--, and NO-3-substituted membranes showed modified B-bands with variously upshifted peak temperatures. The extent of the peak temperature upshift varied in parallel with the extent of inhibition of O2 evolution depending on the anion species. A normal EPR S2 multiline signal was induced in Br--substituted membranes, but its amplitude was reduced to less than 10% in F--, NO-3-, CH3COO--, and SO2-4-substituted membranes, In contrast, the g = 4.1 signal from S2 was markedly enhanced in F-- and NO-3-substituted membranes, not much affected in CH3COO-- and SO2-4-substituted membranes, and decreased to 70% in Br--substituted membranes. Based on these data, the effect of various types of S2 modification on the O2-evolving activity was discussed. It was suggested that anions have an important role in regulating the interaction between the Mn atoms, and thereby adjust the redox properties of the S2 state to enable further transitions beyond S2. PMID- 3039917 TI - Manganese poisoning and the attack of trivalent manganese upon catecholamines. AB - Human manganese poisoning or manganism results in damage to the substantia nigra of the brain stem, a drop in the level of the inhibitory neurotransmitter dopamine, and symptoms resembling those of Parkinson's disease. Manganic (Mn3+) manganese ions were shown to be readily produced by O-2 in vitro and spontaneously under conditions obtainable in the human brain. Mn3+ as its pyrophosphate complex was shown to rapidly and efficiently carry out four electron oxidations of dopamine, its precursor dopa (3,4-dihydroxyphenylalanine), and its biosynthetic products epinephrine and norepinephrine. Mn3+-pyrophosphate was shown to specifically attack dihydroxybenzene derivatives, but only those with adjacent hydroxyl groups. Further, the addition of Mn2+-pyrophosphate to a system containing a flux of O2- and dopamine greatly accelerated the oxidation of dopamine. The oxidation of dopamine by Mn3+ neither produced nor required O2, and Mn3+ was far more efficient than Mn2+, Mn4+ (MnO2), O2-, or H2O2 in oxidizing the catecholamines. A higher oxidation state, Mn(OH)3, formed spontaneously in an aqueous Mn(OH)2 precipitate and slowly darkened, presumably being oxidized to MnO2. Like reagent MnO2, it weakly catalyzed dopamine oxidation. However, both MnO2 preparations showed dramatically increased abilities to oxidize dopamine in the presence of pyrophosphate due to enhancement of the spontaneous formation of the Mn3+ complex. These results strongly suggest that the pathology of manganese neurotoxicity is dependent on the ease with which simple Mn3+ complexes are formed under physiological conditions and the efficiency with which they destroy catecholamines. PMID- 3039918 TI - Effect of halothane on the Ca2+-transport system of surface membranes isolated from normal and malignant hyperthermia pig skeletal muscle. AB - Trapezius muscle from normal and malignant hyperthermia (MH) pigs was used to investigate the effects of halothane on contractile properties and on the calcium transport system of isolated surface membranes. We observed that (i) halothane, diluted in dimethyl sulfoxide, induced a higher isometric contracture response in MH muscle than in normal muscle, (ii) halothane had a more pronounced inhibitory effect on the sarcolemmal Ca2+-ATPase activity in MH membrane, and (iii) the actively accumulated calcium was released in higher amounts in MH muscle than in normal muscle. These results suggest that halothane might induce, in vivo, an important influx of extracellular calcium ions through the MH sarcolemmal membranes and this pool of intracellular calcium may constitute the trigger for the defective sarcoplasmic reticulum "calcium-induced calcium-release" system. PMID- 3039919 TI - [Serum NSE (neuron-specific enolase) levels in small cell lung cancer, with special reference to the measurement of serial serum NSE levels]. AB - Serum neuron-specific enolase (NSE) was measured in 105 patients with lung cancer without prior therapy. Increasing serum levels of NSE were observed in 84% of cases of small cell lung cancer (SCLC). 11% of adenocarcinomas, 18.5% of squamous cell carcinomas and 37.5% of large cell carcinomas. Serum NSE levels changed in parallel with the effect of initial treatment. Although two of 5 patients with SCLC, in whom serial NSE levels were measured after treatment, showed elevation of serum NSE levels before clinical relapse, elevated serum NSE levels were not observed in 3 patients at clinical relapse. These data seem to suggest that measurement of serial NSE is useful for predicting relapse prior to clinical recognition. However, it still remains an important point to carry out clinical general surveillance for relapse. PMID- 3039920 TI - [Studies on the appropriate administration of cisplatin based on pharmacokinetics and toxicity]. AB - Ninety-six patients with non-small cell lung cancer were treated with cisplatin (80-150 mg/m2). The pharmacokinetics of cisplatin were studied in 27 of these patients. Cisplatin was administered intravenously for 30-120 min. Plasma concentrations of total platinum and ultrafilterable (non-protein-bound) platinum were monitored by flameless atomic absorption spectrophotometry. Maximum total platinum levels in plasma were attained at the end of infusion, and thereafter decayed in a biphasic fashion, with an initial phase half-life (t1/2 alpha) of 10.2 to 14.6 min and a secondary phase half-life (t1/2 beta) of 41.7 to 81.3 h. The half-life was prolonged as the dosage was increased. Non-protein-bound platinum was rapidly cleared to below the measurable level within 4 hours at 80 mg/m2 and 120 mg/m2 of cisplatin, but declined in a biphasic manner, with t1/2 alpha of 31.2 min and t1/2 beta of 20.1 h at 150 mg/m2 of cisplatin. Toxicities were increased according to dosage, and decreased with a 50 mg/m2 X 3 days regime. Nephrotoxicity and ototoxicity were the dose-limiting factors in the single-dose escalation scheme used. In conclusion, the optimal dosage of cisplatin appeared to be 120 mg/m2 considering its toxicity. PMID- 3039921 TI - [A phase II study of vindesine for pretreated non-small cell lung cancer]. AB - A phase II evaluation of vindesine (VDS) was performed in 16 patients with non small cell lung cancer (ten patients with adenocarcinoma, six patients with squamous cell carcinoma, and one patient with large cell carcinoma). All except one of the patients had had prior chemotherapy. VDS at a dose of 3 mg/m2 was given intravenously every week for more than three weeks. Among 16 evaluable patients, two patients with pretreated adenocarcinoma of the lung showed partial response. The response rate for VDS was 12.5%. Toxic effects included leukopenia (94%), anemia (44%), thrombopenia (13%), alopecia (38%), peripheral neurotoxicity (38%), liver injury (19%), constipation (13%), anorexia (13%), nausea (13%), stomatitis (6%) and fever (6%). PMID- 3039922 TI - [Microangiopathic hemolytic anemia (MAHA) and renal failure induced by anti neoplastic agents--a case report]. AB - A 65-year-old man underwent left-upper lobectomy for large cell carcinoma of the lung on November 8, 1984 (pT1N0M0: Stage I a). He was treated with MMC, Futraful, CDDP and CPM as adjuvant chemotherapy. In April 1985, he was re-admitted to our hospital because of progressive dyspnea. He was diagnosed as having drug-induced interstitial pneumonia, and so steroid therapy was started. In July 1985, he suffered from anemia, thrombocytopenia, proteinuria and azotemia progressively, and died due to pulmonary hemorrhage and edema. At necropsy, no cancer recurrence was found. It thus seemed that the cause of death was microangiopathic hemolytic anemia and renal failure induced by anti-neoplastic agents. PMID- 3039923 TI - [A case of gastric cancer in which long-term administration of 5'-deoxy-5 fluorouridine (5'-DFUR) proved effective]. AB - A patient with gastric cancer (mucinous adenocarcinoma), classified as Borrmann Type 4, refused surgery and a chemotherapy regimen was applied. 5'-DFUR (5'-deoxy 5-fluorouridine) was administered orally at 1,200 mg/day for 23 weeks, for a total dosage of 110.8 g. The patient showed marked remission: the tumor nearly disappeared, distensibility reappeared from the central region of the corpus ventriculi to the incisura angularis, and only small protrusions classified by Yamada's criteria as Type I, remained on the anterior and posterior wall and pars pylorica of the lesser curvature. Side effects were slight anorexia and diarrhea. These could be avoidable by temporal interruption of administration or dosage reduction, to allow long-term treatment. This case can be considered to be one example in which 5'-DFUR proved effective against gastric cancer. PMID- 3039924 TI - [The effect of etoposide (Et) on primary hepatocellular carcinoma]. PMID- 3039926 TI - Parvovirus associated aplastic crisis in homozygous sickle cell disease. AB - Aplastic crises in homozygous sickle cell disease in Jamaica predominantly affect children and occur in epidemics. Of 67 cases in a cohort study of 314 children with homozygous sickle cell disease, 62 were attributable to human parvovirus infection. Affected children were aged 0.5-12.5 years, and the incidence rose to 28% by 10 years. No recurrences were seen. Symptoms and signs on presentation were attributable to the viraemia and acute anaemia. Asymptomatic thrombocytopenia was common. Blood transfusion was given in 54 cases (87%). Thirty eight children (61%) were admitted to hospital, 16 of whom were extremely ill on presentation and one of whom died soon after admission. Twenty four (39%) were managed as outpatients, 16 of whom were transfused. Parvovirus associated aplastic crisis is a self limited condition with excellent prognosis if diagnosed promptly and managed appropriately. PMID- 3039925 TI - Efficacy of varicella vaccine in patients with solid tumours. AB - Thirty nine children with malignant disease and without antibodies to varicella zoster virus were immunised with a live Oka strain varicella vaccine. Seroconversion was shown in 24 of those who were vaccinated but five of 18 who responded to the vaccine who were followed up for over six months subsequently lost their antibodies. Of 10 children who were revaccinated, nine responded to the second dose but three lost their antibodies within six months to two years. Four children developed reactions related to the vaccine, one local and three generalised, but all these were transitory and of no clinical concern. Nine of those who were vaccinated, including four who did not respond to the vaccine, have subsequently had close exposure to varicella but none has developed the disease. PMID- 3039927 TI - Retroviruses. PMID- 3039928 TI - Human retroviruses and paediatric disease. PMID- 3039929 TI - Effect of peripheral benzodiazepine receptor agonist (RO 5-4864) on inotropism in isolated rat atria. PMID- 3039930 TI - [1 case of angiomatoid fibrous malignant histiocytoma]. PMID- 3039931 TI - Differential diagnosis of chordoma immunohistochemical aspects. PMID- 3039932 TI - Malignant mixed mesodermal tumors of the ovary. A clinico-pathological study of four new cases. PMID- 3039933 TI - [Intracranial melanotic progonoma. Report of a case with an immunohistochemical and ultrastructural study]. PMID- 3039934 TI - [The effects of low-dose continuous infusion of dibutyryl cyclic AMP (DBcAMP)]. PMID- 3039935 TI - [Current role of radionuclide imaging in pediatric cardiology]. AB - Three main nuclear medicine methods are used in paediatric cardiology: sequential first-pass radionuclide imaging of the cardiac cavities, radionuclide equilibrium ventriculography and radionuclide myocardial imaging. Valuable functional information is obtained, and invasive explorations can be avoided in an ever increasing number of cases. Of particular interest is left-to-right shunt measurement which indicates that atrial septal defects must be surgically corrected when the pulmonary/systemic flows ratio (QP/QS) is above 2. This technique is also useful to evaluate the tightness of repairs in ventricular and atrial septal defects. Radionuclide studies of the right and left ventricles may detect dysfunction in one or the other cavity. The left ventricular ejection fraction is reduced in myocardiopathy an in aortic or mitral valve diseases seen at a late stage. The right ventricular function is often abnormal, notably during exercise, after repair of the tetralogy of Fallot and after atrial correction of complete transposition of the great arteries. An altered ejection fraction in patients with single ventricle is also a sign of deterioration. Right ventricular diastolic overload evaluated by radionuclide equilibrium ventriculography correlates with the QP/QS ratio value in atrial septal defects and with the inducibility of ventricular tachycardia by endocavitary pacing in repaired tetralogy of Fallot. Thallium 201 myocardial imaging provides information on myocardial ischaemia, notably that associated with congenital abnormalities of the coronary arteries. Its use had now been extended, albeit with some limitations, to the evaluation of right ventricular systolic overload. Other radionuclide techniques are being developed with new tracers: Kryton 81m for studies of the right ventricle, short-lived radionuclides for first-pass studies, Iodine 123-labelled fatty acids for myocardial imaging. More recently, some substrates, such as deoxyglucose, have been labelled with positron emitters permitting in vivo metabolic studies. PMID- 3039936 TI - Lack of redox control of the anaerobically-induced nirB+ gene of Escherichia coli K-12. AB - Operon fusion strains of Escherichia coli K-12 have been used to demonstrate that transcription of the structural gene for NADH-dependent nitrite reductase is regulated by oxygen repression and induction by its substrate, nitrite. This two stage regulation of nirB is totally dependent upon a functional Fnr protein. Unlike the Fnr-dependent fumarate reductase operon, nirB transcription is not repressed by nitrate. These results suggest that the Fnr protein is simply a positive control protein essential for the derepression of some, but not all, anaerobically-induced operons rather than a more general redox-sensitive regulator, as suggested by the redox control hypothesis for the regulation of gene expression in facultatively anaerobic bacteria. PMID- 3039937 TI - [Adoptive immunotherapy in glioblastoma multiforme: experience with the use of intrathecal infusions of autologous leukocytes]. PMID- 3039938 TI - Complications in gastric bypass. PMID- 3039940 TI - Application of ESR spectroscopy in toxicology. AB - Spin trapping in vivo was first achieved in the author's laboratory and is shown to be a feasible method for demonstrating that highly reactive free radical intermediates are generated in the tissues of intact animals as a result of the exposure to certain toxic compounds and to ionizing radiation. The method is based on the property of spin trapping agents (nitrones) to react readily with reactive free radicals to produce stable radical adducts at the site of their origin in target organs. The radical adducts can then be detected by electron spin resonance spectroscopy to determine the intensity of radical production (i.e., number of radicals which were trapped), and, in most cases, identify the nature of the radical that was produced. The type of spin trapping agent employed determines the type of radicals which can be trapped and, at this stage of development of the technique, the number of useful in vivo trapping agents is rather limited. PMID- 3039939 TI - Tetrachloromethane metabolism in vivo under normoxia and hypoxia. Biochemical and histopathological effects relative to alkane exhalation. AB - About 250 mumol/kg tetrachloromethane is metabolized by male Sprague-Dawley rats receiving a dose of 500 mumol/kg by inhalation. Determination of enzyme activities and histopathological assessment show that various parameters reflecting cellular injury do not differ for the same amount of tetrachloromethane metabolized under different oxygen partial pressures. On the other hand, the amounts of ethane and pentane exhaled under hypoxic conditions are greater than under normoxia. The previously reported differences in the toxicity of tetrachloromethane upon exposure under normoxia or hypoxia seem to be mainly due to the different amounts of tetrachloromethane metabolized under both conditions. PMID- 3039941 TI - Detection of oxygen activation and determination of the activity of antioxidants towards reactive oxygen species by use of the chemiluminigenic probes luminol and lucigenin. AB - Reactive oxygen metabolites can transfer, via oxygenation, the chemiluminigenic probes luminol and lucigenin to an excited state and thus induce light emission from these probes. This technique has been applied to a study of the effect of common food antioxidants on the availability of reactive oxygen species in aqueous model systems and in microsomal preparations. In all systems tested, propyl gallate and its congener octyl gallate proved to be the most active scavengers of the oxygenating species involved in the chemiluminescence reaction. In aqueous model systems, butylated hydroxyanisole was 10 times less efficient than the gallic acid ester antioxidants. In biological material, the lower scavenger activity of butylated hydroxyanisole is further compromised by its ability to induce a marked increase in production of hydrogen peroxide in the endoplasmic reticulum. Butylated hydroxytoluene is only marginally active in aqueous media and in biological material. It is concluded that gallic acid ester antioxidants may possess a significant protective potential towards toxic and carcinogenic agents due to their ability to remove toxic oxygen metabolites. PMID- 3039943 TI - The effects of ultraviolet irradiation of the skin on herpes simplex virus infection: alteration in immune function mediated by epidermal cells and in the course of infection. AB - Previously, we demonstrated that Ia+ epidermal cells (EC) have herpes simplex virus (HSV) antigen-presenting capacity in vitro and play an important role in resistance to HSV infection in vivo. In the present study, we investigated the effects of in vivo ultraviolet (UV) irradiation of the skin on the HSV-immunity function of EC both in vitro and in vivo and on the pathogenesis of HSV infection. Immune T cells cultured with EC and HSV antigen showed a proliferative response in vitro. Exposure of the skin to UV light 1 to 3 days before preparation of EC resulted in dose-dependent impairment of this proliferation. This UV-induced impairment of the accessory cell function of EC was accompanied by a parallel reduction of the number of Ia+ EC. We also transferred these EC stimulated T cells to intracutaneously infected nude mice. Immune T cells stimulated with EC obtained from irradiated mice did not effectively clear HSV and allowed development of zosteriform skin lesions. In contrast normal-EC stimulated immune T cells completely prevented the formation of a zosteriform rash. In addition, mice irradiated with UV on shaved midflank skin 2 days before intracutaneous inoculation of HSV showed increased severity of infection and a higher incidence of latency compared with control mice. These studies indicate that in vivo UV irradiation of the skin abrogates the immune function of EC both in vitro and in vivo, and affects HSV pathogenesis. The implication of our results for the better understanding of the effect of UV on acute and recurrent HSV infections is discussed. PMID- 3039942 TI - Relationship of basic research in toxicology to environmental standard setting: the case of polybrominated biphenyls in Michigan. AB - The accidental contamination of dairy cattle feed in Michigan in 1973-74 with polybrominated biphenyls (PBB) led to the contamination of cattle and people consuming their products. This led to an extensive animal and product monitoring and disposal program conducted by the Michigan Department of Agriculture and the Department of Natural Resources. It also led to several studies of the people of Michigan, extensive research on the chemicals, and an unprecedented establishment by the Legislature of a Toxic Substance Control Commission. Only a few relatively minor components of the PBB mixture that contaminated Michigan are metabolized and another group of minor components seem responsible for the toxicity, which, similar to that caused by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), includes induction of microsomal enzymes, liver hypertrophy, thymic involution, porphyria, anorexia and chloracne. PBB were found to produce the "dioxin-like" toxicity with roughly 0.01% the potency of TCDD. Both non-toxic as well as toxic congeners were found to be tumor promotors. To date it is impossible to unequivocally conclude that any human health effects can be attributed to PBB. The Toxic Substance Control Commission was established as an independent oversight body with responsibility to gather information, investigate, coordinate and make recommendations concerning toxic substances and the handling of toxic substances incidents. The Commission has declared two toxic substances emergencies and made several recommendations for regulating and solving toxic substances problems but its major activities have evolved towards a role as an environmental ombudsman. PMID- 3039944 TI - Hemagglutination with transmissible gastroenteritis virus. AB - Transmissible gastroenteritis virus grown in primary swine kidney cell cultures agglutinated erythrocytes from chicken, guinea pig and cattle but not erythrocytes from mouse and goose. The optimal incubation temperature was at 4 degrees C. The hemagglutination (HA) reaction was inhibited by specific antiserum. Some factors involved in the HA and HA-inhibition (HI) were investigated and standard HA and HI tests were established. HI antibody titers of individual pig sera showed a significant positive correlation with their neutralizing antibody titers. PMID- 3039945 TI - HSV-1 virulence for mice by the intracerebral route is encoded by the BamHI-L DNA fragment containing the cell fusion gene. AB - The phenotype of pathogenicity by direct intracerebral inoculation of herpes simplex virus type 1 (HSV-1) was mapped in the viral genome. This phenotype could be rescued by cotransfection of unit length HSV-1 DNA of an avirulent strain with the BamHI fragment L (0.70-0.738 map units) cloned from a virulent strain. The virulence function was localized in the 2.0 Kb NruI-BamHI fragment in the right hand side of BamHI-L, the same region that encodes a virus cell-fusion gene (3). Transduction of virulence was linked with the phenotype of a larger plaque size. It is concluded that a neurovirulence function resides in the BamHI-L fragment of the HSV-1 genome, closely linked to the viral gene for cell fusion. PMID- 3039946 TI - Macromolecular synthesis at the early stage of herpes simplex virus type 2 (HSV 2) latency in a human neuroblastoma cell line IMR-32: repression of late viral polypeptide synthesis and accumulation of cellular heat-shock proteins. AB - We have shown that a latent infection of herpes simplex virus type 2 (HSV-2) can be established in a human neuroblastoma cell line IMR-32 if the infected cells are cultured at 40 degrees C. In the present study, viral polypeptides and cellular heat-shock proteins which were synthesized in HSV-2 infected IMR-32 cells cultured at 40 degrees C were analyzed by polyacrylamide gel electrophoresis. It was found that the synthesis of late viral polypeptide ICP 5 was markedly reduced in the infected cells at 40 degrees C as compared with those at 37 degrees C. Although infection of IMR-32 cells with HSV-2 at 40 degrees C resulted in shutoff of cellular protein synthesis, it was found that some cellular heat-shock proteins (90, 72 and 70 kd polypeptides) were synthesized and accumulated intracellularly. These findings suggest that modification of cascade regulation of HSV-2 polypeptide synthesis and/or accumulation of heat-shock proteins may be involved in the incomplete arrest of virus growth and in survival of the infected cells, leading to the establishment of HSV-2 latency in IMR-32 cells. PMID- 3039947 TI - Inhibition of porcine parvovirus replication by empty virus particles. AB - The influence of empty porcine parvovirus (PPV) particles on viral replication was examined in cell cultures and in swine. Following extensive purification, homogeneous preparations of full and empty PPV preparations were obtained and used for in vitro and in vivo analyses. In the first in vitro experiment, swine testes cells were infected with mixtures of various ratios of empty and full (E/F) particles. The production of both intracellular and extracellular virus was markedly inhibited in the presence of empty particles. This inhibition was dependent upon the concentration of empty particles present in the mixture. In the second in vitro study, various concentrations of empty particles were added prior to full virus infection. Again, marked inhibition of progeny virus production was evident and related to the concentrations of empty particles added. Based on the results of in vitro studies, the influence of empty particles on PPV infection in swine was tested by infecting mid-term and late-term gestation swine fetuses with various E/F particle ratios. Both mid-term and late term fetuses exposed to 0:1, 1:1 and 5:1 E/F ratios displayed gross pathological evidence of PPV infection whereas fetuses exposed to E/F ratios of 30:1 or greater were grossly normal in appearance. However, fetuses infected with 30:1, 50:1 and 300:1 E/F ratios showed evidence of virus in their tissues by DNA hybridization. Regardless of the E/F ratios, late-term infected fetuses responded with high antibody titers ranging from 1024 to 4096. The results from these studies suggested that empty particles interfered with viral replication in both cell culture and in animals. PMID- 3039948 TI - Effect of platelet-derived factor on expression of bovine leukemia virus genome. AB - Plasma of cattle infected with bovine leukemia virus (BLV) contains a factor, plasma blocking factor (PBF), that inhibits the expression of viral genome in cultured lymphocytes from BLV infected cattle. When platelet lysate was added to this culture, BLV antigen became detectable in the culture and there are some factors (PDF) in platelet lysate which have inhibitory activity against PBF. The PDF was present in platelet from BLV-free cattle as well as BLV-infected cattle at relatively high titer. The effect of platelet lysate against PBF on the expression of BLV genome seemed to be irreversible. PMID- 3039950 TI - [Cancer of the jejunum and Fraumeni syndrome]. AB - An observation of multiple primary tumors of various histogenesis and a rare combination (uterine body carcinoma, mammary fibroadenoma, bronchial carcinoid, jejunum carcinoma) developing in a female patient at various periods of her life is described. The autosome-dominant type of the transfer proves the possibility of a genetic predisposition of the tumor development (the patient's mother and brother also had tumors). This observation demonstrates the variety of tumor combinations and tumor topography in cases of genetically predetermined blastomogenic process. This may represent a basis for distinguishing separate variants of Fraumeni syndrome. PMID- 3039949 TI - [Various aspects of the morphology of granular cell tumors]. AB - Histology and ultrastructure of human granular cell tumors (GCT) are studied. Apart from "granular" cells which are the main component of the tumor, GCT contain the cells of ordinary structure with signs of neurogenic differentiation, cell processus typical for the nervous tissue and peripheral nervous tissue tumors (these processus are in a close space connection with "granular" cells) as well as structurally transitional cells. This implies that "granular" cells are likely to originate from the ordinary lemmocytes as a result of disturbance and hypertrophy of intracellular processus. Preoperative diagnosis of this rare peculiar neurogenic tumor is difficult and the diagnosis is possible only after the operative material examination. If other signs of malignancy are lacking, infiltrative growth typical for these tumors allows one to consider them locally destructive neoplasms. PMID- 3039951 TI - Quantitative sensory testing demonstrates that subclinical sensory neuropathy is prevalent in patients with cancer. AB - Vibration threshold (VT) determinations were used to assess the function of the large nerve-fiber sensory system in 171 patients with cancer and 58 healthy subjects. Significant differences in VT indicate dysfunction of this sensory system in the cancer group. Twelve percent of the cancer patients had elevated VT compared with 1.7% of control subjects. Elevated VT was not associated with risk factors for neuropathy such as diabetes, renal disease, poor nutrition, or treatment with chemotherapy. Although VT elevation was associated with alcoholism and increasing age, these variables accounted for only a small proportion of the variance in VT. These data suggest that VT determinations are a useful method for quantifying sensory abnormalities in cancer patients. Sensory abnormalities occur in a significant proportion of patients with cancer and seem to be related directly to the neoplasm, rather than to known risk factors for neuropathy. PMID- 3039953 TI - Extended neuralgic amyotrophy syndrome. AB - Neuralgic amyotrophy refers to an idiopathic syndrome where weakness and wasting occur in one limb, usually in the muscles innervated by the upper brachial plexus. Seven patients are presented who developed cranial nerve involvement (facial, spinal accessory) in the midst of a typical attack of neuralgic amyotrophy or who developed either recurrent brachial or brachial and lumbosacral plexopathies. An underlying demyelinating neuropathy was identified in one patient and two patients were herion addicts. These reports confirm that neuralgic amyotrophy may occasionally form part of a more extensive disorder of the peripheral nervous system, thereby providing indirect support for the role of a systemic immunological factor in pathoetiology. PMID- 3039952 TI - Otorhinolaryngological findings in AIDS patients: a study of 63 cases. AB - Patients with acquired immunodeficiency syndrome (AIDS) frequently show signs and symptoms involving the head and neck. We reviewed the clinical histories of 63 cases with AIDS, with patients treated from 1982 to December 1985, in our hospital. Findings referable to the head and neck were seen in 43 patients. Comparison with a previous study in San Francisco revealed remarkable differences. We found otolaryngological manifestations in 68% of the patients in contrast to 41% in this latter study. We saw more rapidly increasing neck masses, a greater incidence of shortness of breath and chronic cough, and an increased occurrence of candidiasis. The number of cases with Kaposi's sarcoma lesions was equal in both studies. PMID- 3039954 TI - A study of arthritis in Papua New Guinea. AB - A study was undertaken of 182 Melanesian patients with arthritis who were admitted to three major hospitals in Papua New Guinea between 1977 and 1982. There were 118 male and 64 female subjects, whose mean ages were 29 years and 25 years respectively. A diagnosis was made in 101 cases (55.5%) but in 81 cases, because assessment and management had been inadequate, a diagnosis could not be made. The commonest cause was infectious arthritis (44%), followed by rheumatoid arthritis (14%), gout (8%), and reactive arthritis (8%). In the 44 subjects with infectious arthritis, gonorrhea was the cause in 25 cases (57%) and in a further six cases (14%) it was attributed to Ross River virus infection. In five cases, arthritis was believed to be caused by Wuchereria bancrofti infection. In several cases, chronic arthritis was present in association with ankylosing spondylitis, psoriasis, systemic lupus erythematosus, dermatomyositis, or systemic sclerosis. PMID- 3039955 TI - The development of Cushing's syndrome from a previously silent pituitary tumour. AB - A 60 year old woman originally presented with headache. Investigations revealed a pituitary tumour and endocrine investigations at that time showed normal plasma cortisol levels. Seven years after removal of this tumour, the patient developed the clinical and biochemical features of Cushing's disease. Immunoperoxidase staining of the original tumour was positive for adrenocorticotrophic hormone. This report suggests that immunocytochemistry may have an important role in the routine evaluation of pituitary tumours. PMID- 3039956 TI - Adult onset Still's disease or coxsackie polyarthritis? PMID- 3039957 TI - 'Benign' pleomorphic adenoma of the parotid. AB - A personal series of 141 surgically treated parotid tumours is reviewed. Of 112 parotid tumours in which a diagnosis of 'benign' pleomorphic adenoma or other benign tumour was made, 8% proved to be malignant on histological examination. Three patients were thought to demonstrate malignant change in long standing 'benign' parotid tumours. It is concluded that in otherwise healthy patients, discrete, mobile parotid tumours should be removed without undue delay because of the risk of misdiagnosis and the chance of late development of malignancy in pleomorphic adenomata. Malignant tumours simulate benign tumours often enough to warrant treating all as potentially malignant. PMID- 3039958 TI - Recurrent pleomorphic adenoma of the parotid. AB - Nineteen patients with recurrent pleomorphic adenoma of the parotid are reported. The aetiology of recurrence is discussed and capsule rupture is emphasized as a major cause, especially of multinodular recurrence. Further surgical attempts to remove recurrences are justified by the possibility of misdiagnosis of the original tumour and the late development of malignancy in recurrent nodules. Tumour clearance was achieved in all but two of 19 patients with recurrence or incomplete primary excision in those in whom it was attempted. Radiotherapy does not seem to be an appropriate alternative to repeated surgical excision. PMID- 3039959 TI - Glomus intravagale tumour: aspects of management. AB - Glomus intravagale tumours are relatively rare tumours. A case is described and some aspects of its management are discussed with emphasis on the pre-operative investigation and diagnosis of this condition. Selective embolization of the lesion before operation is an important adjunct in treatment to reduce the vascularity of the tumour during operation. Intra-operative cardiac arrest due to traction of the vagus nerve was encountered. The methods to avoid this complication are discussed. PMID- 3039960 TI - Virus and virus-like particles in the faeces of cats with and without diarrhoea. AB - Negative staining electron microscopy was used to identify viruses in 166 normal and 62 diarrhoeal faecal samples from 208 cats admitted to an animal shelter during a 16-month period (March 1984 to June 1985). On the basis of size and shape 7 distinct viral types were detected: 24 nm parvovirus-like particles, 30 nm astrovirus, 30 nm picornavirus-like particles, reovirus, rotavirus, coronavirus and a 75 nm "togavirus-like" particle. The incidence of these particles in the 208 cats was 11%, 7%, 6%, 0.4%, 5%, 1% and 1% respectively. Virus isolation studies using 40 of the faecal samples succeeded in isolating reovirus 1 in 2 cases. Immune electron microscope studies demonstrated the presence of antibody in a human serum to cat astrovirus, but failed to clarify the identity of the parvovirus-like particles and picornavirus-like particles, other than showing that some of the parvovirus-like particles were not related to feline panleukopenia virus. It was found that parvovirus-like particles, astrovirus, picornavirus-like particles, reovirus and rotavirus could be excreted by cats with normal faeces as well as cats with diarrhoeal faeces. Parvovirus like particles, astrovirus, picornavirus-like particles and rotavirus could be excreted in high concentration in normal faeces. There was no simple relationship between age and diarrhoea in the population of cats studied. Age was not a critical factor in the excretion of parvovirus-like particles, astrovirus, picornavirus-like particles and rotavirus. The incidence of diarrhoea was not clearly associated with the seasons. PMID- 3039962 TI - Further studies on in vitro and in vivo assays of hemorrhagic enteritis virus (HEV). AB - Isolation of hemorrhagic enteritis virus (HEV) from spleens of infected turkeys in the MDTC-RP19 lymphoblastoid cell line was compared with detection of HEV antigen in the same spleens using the agar gel precipitation (AGP) test. A concordance of 80% was found between the two assays. Virus isolation had a sensitivity of 84% and a specificity of 88% compared with the AGP test. RP19 cells were also susceptible to infection with several other avian adenoviruses, but such infection was easily differentiated from that of HEV by a fluorescent antibody (FA) test. Turkeys required 10(2) tissue-culture-infectious doses (TCID) to develop HE-specific lesions and 10(5) TCID to be killed. On the other hand, as little as 10 TCID of apathogenic HEV protected the poults against challenge with virulent HEV. The enzyme-linked immunosorbent assay (ELISA) for detection of HEV antibody was improved by using virus-infected RP19 cells as antigen. The ELISA appears to be more sensitive than the serum-neutralization test. PMID- 3039961 TI - Experimental studies on Marek's disease in Japanese quail (Coturnix coturnix japonica). AB - Day-old quails experimentally infected with Marek's disease (MD) virus of quail origin developed lymphoid tumors. The severity of the disease increased considerably with serial passage. Tumor transplants could be made with cells derived from gross tumors in skeletal muscles, spleen cells, and blood from MD affected quails. After five to six serial transplants, the tumor could not be transplanted further. Marek's disease tumor-associated surface antigen (MATSA) was demonstrated in lymphoid cells of spleen and peripheral blood lymphocytes of MD-affected quails. The MATSA of quail differed from the MATSA of chicken. Chickens were susceptible to MD virus isolated and propagated in quails. PMID- 3039963 TI - Antinuclear antibody in chickens with reoviral arthritis. AB - Antinuclear antibody (ANA) in chickens infected with reovirus was first detected at 3 weeks postinfection (PI). The antibody titer was greatest at 10 weeks PI (1:2560) and then declined. From 19 to 30 weeks PI, the birds were negative for ANA. The ANA was of both IgG and IgM types. The association between antinuclear factor and reoviral arthritis is discussed. PMID- 3039964 TI - Enteric viral infections of turkey poults: incidence of infection. AB - Four flocks from one commercial market-turkey operation in Ohio were monitored for the presence of enteric viruses. Each flock was sampled at intervals from placement until at least 7 weeks of age; sampling was more frequent in the first 4 weeks of life. The earliest infections detected were astrovirus infections or combination infections of astrovirus and rotavirus-like virus (RVLV) or astrovirus and rotavirus. During the first 4 weeks of life, astrovirus was the most frequently detected virus, followed by RVLV, then rotavirus. These viruses were seldom detected beyond 4 weeks of age. In three of the four flocks, no viruses were detected in samples collected before 6 days of age; in one flock, however, rotavirus and astrovirus were identified from samples collected at 3 days of age. Experimental infection of specific-pathogen-free poults with astrovirus and RVLV produced enteric diseases in poults and demonstrated that astrovirus was shed into the intestinal tract before RVLV. Poults experimentally infected with astrovirus and RVLV displayed clinical signs of diarrhea and upon necropsy exhibited dilated ceca, frothy gaseous intestinal contents, and loss of intestinal tone. PMID- 3039965 TI - Effects of excess vitamin D3 and cage density on the incidence of leg abnormalities in broiler chickens. AB - A 2 X 2 factorial experiment was designed to investigate the effects of excess vitamin D3 and cage density on the incidence and severity of leg abnormalities in broiler chickens. One hundred eighty 1-day-old broiler chicks were randomly divided into densities of either 10 (680 cm2/chick) or 20 (340 cm2/chick) per cage and fed a diet containing 22% protein and 2879 kcal/kg metabolizable energy formulated to meet National Research Council requirements. Two levels of vitamin D3 were incorporated to supply either 400 ICU or 4000 ICU/kg feed. High density and excess vitamin D3 resulted in a significant (P less than 0.05) increase in the incidence of twisted leg. Differences in incidence could not be explained through differences in body weight or feed consumption. However, broilers fed the excess vitamin D3 consumed more feed but gained less body weight, suggesting that metabolic stress may have been involved. High density appeared to increase the severity of the disorders, whereas excess vitamin D3 had no effect on severity. PMID- 3039966 TI - A trivalent antigen for the detection of type I avian adenovirus precipitin. AB - A double immunodiffusion antigen prepared from cell-culture-propagated CELO virus was not capable of detecting precipitin directed against all of the type I avian adenovirus (fowl adenoviruses) isolates tested. However, an antigen pool containing CELO-4, B-3, and IBH-2 (Tipton) fowl adenovirus isolates detected precipitin directed against representative isolates of 10 type I serotypes. Additionally, this antigen pool markedly improved detection of adenovirus field exposure. PMID- 3039967 TI - Replication of infectious bursal disease virus in continuous cell lines. AB - Three mammalian continuous cell lines--MA-104, Vero, and BGM-70--were tested for their ability to support replication of infectious bursal disease virus (IBDV). Selected tissue-culture-adapted vaccine strains and tissue-culture-adapted field isolates of IBDV replicated in the MA-104, Vero, and BGM-70 cells; cytopathic effects were most pronounced in the BGM-70 cells. The cytopathic effects of the viruses in BGM-70 cells and chicken embryo fibroblast (CEF) cultures were similar. Virus-neutralization titers of selected serum samples determined in BGM 70 cultures compared well with those obtained from CEF cultures. PMID- 3039968 TI - Demonstration of rotavirus and rotavirus-like virus in the intestinal contents of diarrheic pheasant chicks. AB - Intestinal content samples from four flocks of pheasant chicks experiencing diarrhea and increased mortality were evaluated using immune electron microscopy and genome electropherotyping techniques. Enteric viruses serologically and electropherotypically indistinguishable from turkey rotaviruses and turkey rotavirus-like viruses (RVLVs) were detected. Rotavirus-only infections, RVLV only infections, and simultaneous rotavirus and RVLV infections were demonstrated in these flocks. PMID- 3039969 TI - Clinical encephalitis following avian encephalomyelitis vaccination in broiler breeder pullets. AB - Twelve-to-fourteen-week-old broiler breeder pullets and cockerels from two different companies were found to have encephalitis 2 weeks after vaccination with avian encephalomyelitis (AE) vaccine. Histologic examination and virus isolation indicated that affected chickens had AE. All chickens had been vaccinated with either one of two serials of AE vaccine approximately 2 weeks before the onset of clinical signs. PMID- 3039970 TI - An outbreak of avian urolithiasis on a large commercial egg farm. AB - A significant outbreak of avian urolithiasis was observed on a large commercial egg farm. From the initial outbreak site (a single laying house), the incidence of urolithiasis slowly spread in the ensuing months to numerous other laying houses. Increasing mortality associated with urolithiasis commenced during late growout to early lay and then leveled off when egg production peaked. At the height of the outbreak, mortality was typically 0.5% per week; 75% of this mortality was due to urolithiasis. The clinical and pathologic features of this condition are described. Both infectious bronchitis virus (IBV) and fowl adenoviruses were isolated from organ homogenates of sampled birds. A clone of the IBV strain was found to induce nephritis in specific-pathogen-free white leghorns. PMID- 3039971 TI - Characterization of a reo-like virus and its isolation from and pathogenicity for parrots. AB - Necropsy of an African Grey parrot (Psittacus erithacus) revealed subcutaneous hemorrhages, multiple foci and microfoci of necrosis in the liver, spleen, bone marrow, and intestinal lamina propria, mild air sacculitis, and epicarditis. A virus, isolated from the liver, was non-enveloped, was polyhedral with a diameter of 84.9 +/- 3.4 nm, possessed a double-stranded RNA genome, and was stable at pH 3.0 for 30 minutes and at 56 C for up to 120 minutes. The virus was propagated in cell culture, purified by limiting dilution, and inoculated into two African Grey parrots. The experimental infections were fatal on the 8th and 9th days postinoculation in the orally and intramuscularly inoculated birds, respectively, and produced hemorrhages and necrotic lesions that recapitulated those of the index case. PMID- 3039972 TI - Gestational trophoblastic disease in Port Moresby, Papua New Guinea. AB - A retrospective study was performed at Port Moresby General Hospital covering the period 1.1.81-31.3.86. There were 37 cases of hydatidiform mole, 2 of choriocarcinoma and 4 of metastasizing gestational trophoblastic disease of unknown histology, During the study period there were 35,630 deliveries in the same hospital. The incidence of hydatidiform mole was found to be 1:963 deliveries. Among the patients with hydatidiform mole 1 death was recorded whilst in the group with choriocarcinoma and unclassified disease there were 4 deaths. Of the patients treated for hydatidiform mole 48.5% made no follow-up visits to the hospital. PMID- 3039973 TI - Antibody responses to early antigens of varicella-zoster virus (VZV) during varicella and zoster. AB - The antibody responses to membrane and early antigens and thymidine kinase of varicella-zoster virus (VZV) were studied in sera during both varicella and zoster by a test with fluorescent antibody to membrane antigen (FAMA), staining the biochemically transformed cells by the immunofluorescent technique and neutralization of virus-specific thymidine kinase activity, respectively. Similar increases in FAMA antibody titers were demonstrated in sera from patients with varicella and zoster. IgM was detected in both groups, but appeared earlier during varicella than during zoster. Furthermore, the antibody titers to early antigens and virus-specific thymidine kinase were higher in patients with zoster than in those with varicella. These data suggest that different types of antibody responses occur during varicella and zoster. PMID- 3039974 TI - Genetic differentiation between laboratory lines of the musk shrew (Suncus murinus, Insectivora) based on restriction endonuclease cleavage patterns of mitochondrial DNA. AB - The cleavage patterns of mitochondrial DNA (mtDNA) by 17 restriction endonucleases were compared between eight lines of musk shrews derived from different wild-caught stocks. Enzymatic digestion by BamHI, PvuII, XbaI, and XhoI showed a cleavage pattern common to all lines that were from five Japanese islands (Nag, Ize, OKI, TKU, and Tr), Bangladesh (BAN), Sri Lanka (SRI), and Java (Bog), and every line lacked cleavage sites for SalI and SmaI. Different cleavage patterns were detected by the remaining 11 enzymes. Within the BAN line, the presence of at least two types of mtDNAs was proved by six enzymes and was not contradictory to the maternal pedigrees going up to the wild ancestors of the stock. More than 30 cleavage sites of the shrew mtDNA were mapped by double digestion methods. Nucleotide diversities of mtDNA were calculated from these maps and were estimated to be less than 0.5% among Japanese and Bog lines but to be 3.8% between BAN and the other seven lines and 2.3% within the BAN line. These results indicate that BAN shrews differentiate from the other lines to the intersubspecific extent reported in mice previously. PMID- 3039975 TI - Retinal rod GTPase turnover rate increases with concentration: a key to the control of visual excitation? AB - Guanosine triphosphate (GTP) binding proteins mediate cellular responses to hormones, neurotransmitters, growth factors and light. Activated GTP binding proteins are shut off by GTPase mediated hydrolysis of GTP. Photoreceptor GTPase rates are reported to be 10-50 times too slow to account for electrophysiological recovery time after light stimulus. Recovery rates of other parts of the system, however, appear fast enough. We present evidence that the GTPase rate increases markedly with photoreceptor membrane concentration implying the existence of a diffusible factor controlling the GTPase. When extrapolated to physiological concentrations, the GTPase turnover rate is fast enough (0.25-1.5 sec) to account for the recovery rate of the light stimulated signal of the photoreceptor cells. PMID- 3039976 TI - Transforming growth factor-beta inhibits Leydig cell steroidogenesis in primary culture. AB - The effects of transforming growth factor (TGF) on Leydig cell steroidogenesis in primary culture were investigated. Basal testosterone levels were 3.7 +/- 0.54 ng/ml (mean +/- SE, N = 7). In the presence of hCG (10 ng/ml), testosterone levels increased to 22.77 +/- 3.05 ng/ml. TGF-beta caused a dose dependent inhibition of hCG-stimulated testosterone formation but without effects on basal levels. TGF-beta also inhibited 8-bromo cyclic AMP-induced testosterone formation and hCG-stimulated cyclic AMP formation. In contrast, TGF-alpha had no effect on either basal or hCG-stimulated testosterone formation and did not modify the inhibitory effect of TGF-beta. Present study indicates that TGF-beta can modulate Leydig cell steroidogenesis. PMID- 3039977 TI - Identification in rat brain of a 19-kDa protein that comigrates on two dimensional electrophoresis with p19, a hormonally regulated phosphoprotein of insulinoma cells. AB - We have previously shown that a set of 19-kDa cytosolic proteins, p19, undergoes hormone-dependent phosphorylation in several peptide hormone-producing tumor cells. Here we show, using comigration on two-dimensional electrophoresis with RIN-1122 rat insulinoma cell p19, that an identical set of 19-kDa proteins is present in rat brain but not in liver or skeletal muscle. We have partially purified p19 from rat brain and have compared the apparent isoelectric variants by tryptic peptide mapping. The data suggest that p19 is a novel phosphoprotein consisting of an unphosphorylated form and of three phosphoforms. PMID- 3039978 TI - Phosphatidyl inositol inhibition of a sperm cyclic AMP-independent protein kinase. AB - Phosphatidyl inositol has been found to inhibit strongly the activity of a cyclic AMP-independent protein kinase located on the external surface of goat epididymal intact spermatozoa. Phosphatidyl inositol at a concentration as low as 10 micrograms/ml inhibited nearly 50% of the ecto-kinase activity for the phosphorylation of the exogenous protein substrate: casein. Phosphatidyl ethanolamine at a relatively high concentration (125 micrograms/ml) inhibited slightly (approx 25%) the activity of the enzyme whereas other phospholipids: phosphatidyl serine and choline, diacyl glycerol, phosphatidic acid and myo inositol-2-phosphate had no appreciable effect on the kinase activity. Phosphatidyl inositol has also served as a potent inhibitor of the phosphorylation of sperm ecto-phosphoproteins by the endogenous kinase activity of intact spermatozoa. By thin layer chromatography it has been shown that the observed inhibitory effect of the phospholipid was not due to any impurities or degraded products of phosphatidyl inositol. Phosphatidyl inositol inhibited the kinase activity noncompetitively with respect to casein and Mg2+ but uncompetitively with respect to ATP. The results raised the possibility that phosphatidyl inositol-mediated high affinity inhibition of protein kinase(s), may constitute a novel mechanism for the regulatory actins of the phospholipid in mammalian cells. PMID- 3039979 TI - Inhibition of bombesin-induced mitogenesis by pertussis toxin: dissociation from phospholipase C pathway. AB - Prior incubation of quiescent cultures of Swiss 3T3 cells with pertussis toxin selectively inhibited the stimulation of DNA synthesis induced by peptides of the bombesin family. While pertussis toxin blocked mitogenesis at an early stage in the action of the peptide, the toxin did not impair the rapid stimulation of polyphosphoinositide breakdown, Ca2+ mobilization or activation of protein kinase C promoted by bombesin. Thus, inhibition of bombesin-induced mitogenesis by pertussis toxin can be dissociated from inactivation of the phospholipase C signalling pathway. PMID- 3039980 TI - Inhibition of cytochrome P-450-linked monooxygenase systems by naphthoquinones. AB - Several naphthoquinones, except 2-hydroxy-1,4-naphthoquinone, were found to inhibit microsomal cytochrome P-450-linked monooxygenase activities in rabbit liver and human placenta. In particular, 5-hydroxy-1,4-naphthoquinone inhibited placental estrogen biosynthesis more effectively than it did hepatic drug oxidation reactions. There was little contribution by superoxide radicals to these enzyme inhibitions by naphthoquinones. Spectrophotometric studies revealed that naphthoquinones bind to the cytochrome P-450 component of the monooxygenase complex in both microsomal systems, suggesting that the inhibition is caused by direct interaction of these compounds with the heme. PMID- 3039981 TI - The inhibitory effect of cyclic AMP on phosphatidylinositol kinase is not mediated by the cAMP dependent protein kinase. AB - Addition of cAMP to cells has been shown to inhibit phosphatidylinositol (PI) metabolism. cAMP has been reported to inhibit an enzyme in this pathway, PI kinase and it has been suggested that this inhibition is due to phosphorylation of PI kinase by the cAMP dependent protein kinase (PKA). In the present study we directly investigated if the inhibitory effect of cAMP was mediated by PKA. In membranes derived from murine hepatocytes we found that cAMP inhibited PI kinase but other adenine derivatives were more potent inhibitors. Moreover, it was found that the effects of the derivatives were unlikely to be due secondarily to the production of cAMP via their interaction with adenosine receptors. Through studies employing an inhibitor of PKA, mutant cells lacking PKA, and addition of purified catalytic subunit of PKA, we found that the inhibitory effect of cAMP was not mediated by PKA. In addition, the inhibitory effect of cAMP and adenosine was retained upon partial purification of PI kinase. Pulse chase experiments affirmed that the inhibitory effect was not due to breakdown of PI but rather to inhibition of its synthesis. We conclude that the inhibitory effect of cAMP and related compounds on PI kinase is not mediated by PKA dependent phosphorylation but rather appears to be a direct effect of these agents. PMID- 3039982 TI - The type I and type II gamma-aminobutyric acid/benzodiazepine receptor: 1. Purification and two-dimensional electrophoretic analysis of the receptor from cortex and cerebellum. AB - The gamma-aminobutyric acid/benzodiazepine receptor complex was purified from rat cortex and cerebellum by benzodiazepine affinity chromatography. Receptors purified from cortex and cerebellum showed different relative affinities for Cl 218872, a non-benzodiazepine ligand which discriminates type I and type II receptors. In contrast, no differences in subunit composition could be detected between these two purified receptor preparations when analyzed by two-dimensional gel electrophoresis. PMID- 3039983 TI - Transforming growth factor beta inhibits Leydig cell functions. AB - The role of transforming growth factor beta (TGF-beta) on the functions of pig Leydig cells cultured in a chemically defined medium was investigated. TGF-beta reduced the number of hCG receptors, without modification of the binding affinity, and reduced the cAMP and testosterone response to this hormone. These effects were dose-dependent. The minimal effective dose was 10 pg/ml (4 X 10(-13) M) and half-maximal inhibition for the three effects was observed at about 100 pg/ml. At maximal effective concentration (1 ng/ml), the inhibitory effect was time-dependent, the first effects were observed after a lag period of 12 h and the maximal effect after 72 h. At maximal concentrations TGF-beta reduced by 70% the number of hCG receptors and the steroidogenic response to this hormone and reduced by 50% the cAMP response to hCG. Moreover, TGF-beta also reduced the cAMP response to forskolin and the steroidogenic effects of this diterpene and 8-Bromo cAMP. In contrast, the conversion of exogenous pregnenolone to testosterone was increased in TGF-beta-treated cells. The inhibition of Leydig cell steroidogenesis can be dissociated from any effect on cell proliferation and is related to modifications located at the membrane level and beyond cAMP formation, but before pregnenolone formation. The results suggest that in the testis, as in other steroidogenic tissues, TGF-beta may play a role in the development and maintenance of differentiated function. PMID- 3039984 TI - The influence of the dT.dG mispair on the activity of the human DNA(cytosine 5)methyltransferase. AB - Synthetic oligodeoxynucleotides containing a dT.dG mispair at a centrally located d(pCG) dimer are methylated at a moderate rate by highly purified human DNA(cytosine-5)methyltransferase (E.C. 2.1.1.37). The presence of a mispaired dT in one strand induced the enzyme to preferentially methylate the opposite strand. PMID- 3039985 TI - Calcium-activated neutral protease inhibitor from rabbit erythrocytes lacks the N terminal region of the liver inhibitor but retains three inhibitory units. AB - Endogenous inhibitors for calcium-activated neutral protease (CANP) were purified from rabbit erythrocytes and liver. The purified inhibitors showed single bands but with significantly different mobilities on sodium dodecylsulfate polyacrylamide gel electrophoresis. Peptide mapping and sequencing analyses have revealed that the erythrocyte inhibitor (429 residues) retains the C-terminal three repetitive units of the liver inhibitor (639 residues), which contains four potential repetitive units for inhibition of CANP. The erythrocyte and liver inhibitors inhibited 3 and 4 moles of CANP on the basis of the molecular weights of 46,000 and 68,000, respectively. PMID- 3039987 TI - Differences in the action of avermectin B1a on the GABAA receptor complex of mouse and rat. AB - The effects of avermectin B1a (AVM) on the gamma-aminobutyric acid (GABA) receptor-chloride ionophore complex of mouse and rat brain were determined using assays of basal and GABA-stimulated 36Cl-uptake by brain vesicles. In the mouse, AVM acted solely as a potent non-competitive inhibitor of GABA-dependent chloride uptake. In the rat, inhibition of GABA-dependent chloride uptake was potent but incomplete, and AVM applied in the chloride uptake medium stimulated chloride uptake in the absence of GABA. The data provide evidence for qualitative differences between the GABA receptor complexes of mouse and rat brain in their responses to AVM. PMID- 3039986 TI - Periodate-oxidized NADP+ is a powerful inhibitor of human placental estradiol-17 beta dehydrogenase. AB - Periodate-oxidized NADP+ inhibits the NAD+-linked activity of human placental estradiol-17 beta dehydrogenase (EC 1.1.1.62). The inhibition appears to be competitive with respect to NAD+ and can be reversed by dialysis or gel filtration. The apparent inhibitor constant for the periodate-oxidized analogue is 0.047 microM. The presence in the incubation mixture of NAD+ protects the enzyme against inhibition. No inhibitory effects of the coenzyme analogue are observed on the NADP+-linked activity of the enzyme. PMID- 3039988 TI - Loss of nerve growth factor receptors in sympathetic ganglia from aged mice. AB - [125I]-Nerve growth factor (NGF) binding was studied in superior cervical ganglia from mice 4-7 months and 24-27 months of age. Scatchard analysis demonstrated losses of both high and low affinity components of NGF receptor. These results indicate loss of NGF receptors may lead to the diminished responsiveness to NGF in aged sympathetic ganglion neurons. PMID- 3039989 TI - D-myo-inositol (1,4)-bisphosphate 1-phosphate. Partial purification from rat liver and characterization. AB - A specific 1-phosphatase acting on myo-inositol (1,4)-biphosphate with a high affinity (Km = 0.9 microM) has been purified 49-fold from soluble proteins of rat liver by anion exchange chromatography followed by gel filtration. This enzyme has a molecular weight of 58,000 as estimated by gel filtration, a pH optimum of 7.5, and requires Mg++ for activity. The only product formed from myo-inositol (1,4)-bisphosphate is the 4-monophosphate. Of 7 other inositol phosphates examined only myo-inositol (1,3,4)-triphosphate was a substrate. PMID- 3039990 TI - Ornithine transcarbamylase deficiency in spf and spf-ash mice: genes, mRNAs and mRNA precursors. AB - Two ornithine transcarbamylase-deficient mice are available, the spf with a variant enzyme and spf-ash with a markedly decreased enzyme protein. Genomic DNA, mRNA and the nuclear precursors for the enzyme in these mutants were analyzed. Southern blot analysis of genomic DNA showed no abnormality in the mutant mice. Blot analysis of hepatic mRNA revealed a slight decrease (67% of control) in spf and a marked decrease (12% of control) in spf-ash; no difference in size was found among the control and the mutant mice (about 1.8 kb). Blot analysis of nuclear mRNA precursors (greater than 25, approximately 9.0 and 4.0 kb) showed no significant difference in size and amount among the control, spf and spf-ash. These results suggest that ornithine transcarbamylase deficiency in the spf-ash results from a mutation, which to some extent affects mRNA processing. PMID- 3039991 TI - Stereospecific recognition sites for [3H]inositol(1,4,5)-triphosphate in particulate preparations of rat cerebellum. AB - A very high density of stereospecific binding sites for inositol-(1,4,5)P3 have been identified in rat cerebellar membranes using [3H]inositol-(1,4,5)P3 and a rapid centrifugation step to separate free and bound ligand. Binding was shown to be rapid and reversible and of relatively high affinity (KD 23 nM). Incubations were carried out at 4 degrees and under these conditions HPLC analysis demonstrated that there was no significant metabolism of [3H]-(1,4,5)P3 in the presence or absence of ATP over 15 min. The specificity of the site has been carefully evaluated using both natural and novel synthetic inositol phosphates. The stereospecificity is very marked with the D-, DL- and L-isomers of Ins(1,4,5)P3 showing a 1:4:2000 ratio of affinity for the binding site. D Ins(2,4,5)P3 was the only other phosphate to show relatively high affinity (KD 1500 nM). HPLC-pure Ins(1,3,4)P3 and Ins(1,3,4,5)P4 were substantially weaker and Ins(1,4)P2, Ins-2-P1, Ins-1-P1, Ins(1,2)-cyclic P1 and inositol were totally inactive at concentrations less than 50 microM. These data are discussed in relation to a putative receptor on the endoplasmic reticulum by which Ins(1,4,5)P3 can initiate the release of bound Ca2+. PMID- 3039992 TI - Receptors for transferrin and transcobalamin II display segregated distribution on microvilli of leukemia L1210 cells. AB - Simultaneous addition of uniform latex particles derivatized with transferrin (0.532 micron) and transcobalamin II (0.345 micron) to leukemia L1210 cells resulted in segregated binding to individual microvilli as demonstrated by scanning electron microscopy. This segregated distribution suggests that individual microvilli are endowed either transferrin or transcobalamin II receptors but not both. Intracellular sorting and segregation of newly synthesized or recycling receptors probably occur prior to expression on the plasmalemma microvilli. PMID- 3039993 TI - Interaction of dimers of inactive enkephalin fragments with mu opiate receptors. AB - Dimeric analogues of the inactive enkephalin fragment Tyr-D-Ala-Gly were synthesized by cross-linking with alkanediamine at the C-terminus. Biological evaluation of these dimers (H-Tyr-D-Ala-Gly-NH)2.(-CH2-)n (DTREn), where n = 0-6, revealed that the fragment inactive for mu receptors was activated by its dimerization, with the maximum activation found with DTRE2, and that the dimer was highly mu-selective. So-called "handicapped" dimers, which lack one of the essential groupings required for enkephalin activity, were found to be far less active, indicating that the dimer interacts bivalently with mu receptors. It seems, therefore, that mu opiate receptors contain at least two equivalent binding sites which are extremely close to each other. PMID- 3039994 TI - O2- photogenerated from aqueous solutions of tetracycline antibiotics (pH 7.3) as evidenced by DMPO spin trapping and cytochrome c reduction. AB - UV-irradiation of several tetracycline antibiotics in aqueous buffer (pH 7.3) resulted in the generation of the superoxide anion radical (O2-) which was detected by cytochrome c reduction and by spin trapping with 5,5-dimethyl-1 pyrroline-N-oxide and was inhibited by superoxide dismutase. A comparison of the O2- yields from the tetracyclines examined showed the trend chlortetracycline (CTC) greater than oxytetracycline (OXY) greater than demeclocycline (DEM) much greater than (doxycycline (DOXY) = tetracycline (TC) = minocycline (MINO) = 0). This trend is in reasonable agreement with clinical reports that CTC, OXY and DEM are potent photosensitizers, TC is only weakly phototoxic whereas MINO is not. These findings suggest that the O2- production may be involved in tetracycline induced phototoxicity. While the two methods for O2- detection gave comparable results for most of the tetracyclines, the spin trapping technique was clearly superior for DOXY which reduced cytochrome c in the dark. PMID- 3039995 TI - Degradation of proteins with blocked amino groups by cytoplasmic proteases. AB - The effect of blocking amino groups on the susceptibility of BSA and calmodulin to high molecular weight protease (HMP) and calpain, the two major cytosolic proteases, was studied. Both proteases hydrolyzed methylated vs. unmodified BSA more slowly. Methylation of BSA resulted in the accumulation of proteolytic intermediates, especially of larger sizes. However, similar fragments were generated from unmodified BSA indicating that rates of hydrolysis rather that sites of proteolytic cleavage were altered. Calmodulin from Dictyostelium discoideum was hydrolyzed rapidly by HMP whereas brain and muscle calmodulins which have a epsilon-N-trimethyl residue on the single surface lysine were relatively stable. PMID- 3039996 TI - Stimulation of ACTH release by human interleukin-1 beta, but not by interleukin-1 alpha, in conscious, freely-moving rats. AB - Ever since two distinct molecules of human interleukin-1 (termed interleukin-1 alpha and interleukin-1 beta) were cloned, sequenced and expressed, it has been a matter of investigation whether these two forms of interleukin-1 possess an identical spectrum of biological activities. Our current studies of interleukin-1 and its involvement in the hypothalamic-pituitary-adrenal axis have indicated that there is a clear-cut differential response to interleukin-1 alpha and interleukin-1 beta. The intravenous injection of human recombinant interleukin-1 beta significantly increased the plasma levels of adrenocorticotropic hormone in a dose-related manner, whereas interleukin-1 alpha did not. This observation suggests for the first time that the two members of the interleukin-1 family may have a different spectrum of biological actions. PMID- 3039997 TI - Animal viruses promote the entry of polysaccharides with antiviral activity into cells. AB - Charged polysaccharides such as heparin and carrageenan show potent antiviral activity in cultured cells. Labelled [3H]heparin binds to several virion particles as evidenced by Sepharose 6B chromatography. This binding is inhibited by carrageenan. The complex [3H]heparin-HSV1 virions is able to enter cells. Thus, almost no entry of [3H]heparin is observed in control HeLa cells, whereas in the presence of HSV1 or poliovirus the amount of radioactivity internalized is enhanced. This internalization is inhibited by carrageenan, suggesting that these two sulphated polysaccharides bind to the same sites on virion particles and may share a similar antiviral mechanism of action. PMID- 3039998 TI - Apocytochrome-c competes with pre-ornithine carbamoyl transferase for transport into mitochondria. AB - Apocytochrome c, the cytosolic precursor of cytochrome c, competes with the precursor of ornithine carbamoyltransferase (OCT) for entry into isolated rat liver mitochondria. PMID- 3039999 TI - Mechanisms of the mAb ALB6(CD9) induced human platelet activation: comparison with thrombin. AB - A CD9 monoclonal antibody described to aggregate human platelets was studied on different platelet functions in order to determine its mechanism of action. After a lag phase of 35 sec the mAb ALB6 induced a transient decrease in 32P polyphosphoinositides, synthesis of 32P-phosphatidate (PA), phosphorylation of myosin light chain (P20) and of 43 KDa protein (P43) and the release reaction. Final biological and metabolic effects of ALB6 thus appear similar to that of thrombin but three differences bring additional information: (i) the lag phase, (ii) the kinetic of ALB6-induced release is identical for all granules whereas the release of dense granules is faster when induced by thrombin. (iii) no external Ca++ is required for ALB6 induced-activation. PMID- 3040001 TI - Mechanism of inhibition of herpes simplex virus replication by delta 7 prostaglandin A1 and delta 12-prostaglandin J2. AB - We studied the effect of prostaglandins (PGs) A1, delta 7-A1, A2, D2, E1, E2, F2 alpha, J2 and delta 12-J2 on the replication of herpes simplex virus type 2 (HSV 2). Of nine PGs we tested, delta 7-PGA1 was found to have the most potent inhibitory effect; 50% inhibitory dose (ID50) was 0.35 microgram/ml in the plaque reduction assays and HSV-2 induced protein synthesis was strongly suppressed at 0.5 microgram/ml whereas at this dose, the protein synthesis of uninfected cells was not inhibited. Dot blot hybridization analysis revealed that delta 7-PGA1 and delta 12-PGJ2 inhibited the primary transcription of HSV-2. Thus we suggest that those PGs are primarily active at the level of mRNA synthesis. PMID- 3040000 TI - Phospholipase C activation and diacylglycerol kinase inactivation lead to an increase in diacylglycerol content in spontaneously hypertensive rat. AB - Activities of three kinases, phosphatidylinositol (PI), phosphatidylinositol 4 phosphate (PIP), and diacylglycerol (DG) kinases, and phospholipase C were measured in erythrocyte ghosts from spontaneously hypertensive rats (SHR) and normotensive Wistar Kyoto rats (WKY). PI kinase activity was significantly higher in SHR than WKY but there was no significant difference in PIP kinase activity between SHR and WKY. The activity of phospholipase C, which hydrolyzes PIP2, was also increased in SHR. However, DG kinase activity was, on the contrary, decreased in SHR. These results suggest that there is a tendency to accumulate DG in SHR. Indeed, DG content in erythrocytes of SHR increased 1.7-fold compared to that of WKY. Such DG accumulation may cause the sustained activation of protein kinase C in SHR, since DG is a physiological activator for protein kinase C. PMID- 3040002 TI - Variation in the molecular mass of the Ah receptor among vertebrate species and strains of rats. AB - The Ah receptor in eight vertebrate species was characterized by labeling the cytosolic fraction of tissue with the photoaffinity ligand, [125I]-2-azido-3-iodo 7,8-dibromodibenzo-p-dioxin, and analysis of the products by denaturing gel electrophoresis. The apparent molecular mass of the dominant labeled peptide showed appreciable species variation: mouse-95 kDa; chicken (embryo)-101 kDa; guinea pig-103 kDa; rabbit-104 kDa; rat-106 kDa; human-106 kDa; monkey-113 kDa, and hamster-124 kDa. Seven inbred strains of rats, had a Ah receptor ligand binding peptide of 106 kDa; however outbred Long-Evans rats were shown to be polymorphic expressing a 101 kDa and/or 106 kDa allelic forms. The notable frequency of structural variation in the Ah receptor is in contrast to the analogous highly conserved steroid hormone receptors. PMID- 3040003 TI - The effect of DMSO on natural DNA conformation in enhancing transcription. AB - In the presence of 5% dimethylsulfoxide, 2.5 fold increase in vitro RNA synthesis rate by E. coli RNA polymerase was observed as superhelical pBR322 DNA was used as template. The effect of DMSO on DNA conformation was studied by: [i] measuring the dissociation constant of EtBr-DNA complex and the maximum EtBr binding site of EtBr in DNA which changed from 7.52 X 10(-7) M and 0.18 site per nucleotide to 9.61 X 10(-7) M and 0.156 site per nucleotide respectively when 10% DMSO was supplemented; and [ii] the increase of average linking number of pBR322 DNA in DMSO solution with topoisomerase I reaction. These results suggest that the stimulation of RNA synthesis may be caused by the easier initiation and elongation of RNA synthesis at some locally loosen regions of DNA affected by another torsionally more twisted counterpart regions in the DMSO microenvironment. PMID- 3040004 TI - GABA-stimulated 36Cl- influx into reconstituted vesicles with purified GABAA/benzodiazepine receptor complex. AB - Solubilized and Purified gamma-aminobutyric acid (GABA)A receptors from membrane vesicles of the bovine cerebral cortex were reconstituted into phospholipid vesicles and 36Cl- influx into the vesicles was examined. GABA induced a significant stimulation of the 36Cl- influx into reconstituted vesicles with 1.5% CHAPS/0.15% asolectin solubilized receptor and flunitrazepam further enhanced the GABA-stimulated influx. The purification of GABAA/benzodiazepine receptor complex and Cl- channel solubilized by 1.5% CHAPS/0.15% asolectin from membrane vesicles was achieved by 1012-S affinity column chromatography. The reconstituted vesicles with the purified receptor complex and Cl- channel also exhibited GABA-stimulated 36Cl- influx. This GABA-stimulated influx of 36Cl- was also enhanced by flunitrazepam, while suppressed by bicuculline, a GABAA receptor antagonist. These results strongly suggest that GABAA receptor is directly coupled with Cl- channel, whereas benzodiazepine receptor may be functionally coupled with GABAA receptor and modulates the GABA-stimulated Cl- influx through GABAA receptor. The present results also indicate that the purified GABAA receptor complex is coupled with Cl- channel and possesses functional characteristics as GABAA receptor. PMID- 3040005 TI - Transmembrane receptor cross-talk: concurrent VIP and alpha 1-adrenergic activation rapidly elevates pinealocyte cGMP greater than 100-fold. AB - The transmembrane regulation of cGMP accumulation, which is poorly understood, was studied using isolated rat pinealocytes. It was found for the first time that VIP stimulates cGMP accumulation several-fold. This stimulation was amplified by phenylephrine acting via alpha 1-adrenoceptors, resulting in a greater than 100 fold increase in cGMP accumulation. These results raise the possibility that cGMP accumulation in other tissues might be regulated by VIP, and that the stimulating effects of VIP might be markedly amplified by catecholamine transmitters in these tissues. It is also possible that other pairs of receptors might control large changes in cGMP in the central nervous system through parallel mechanisms. PMID- 3040006 TI - Mapping structurally perturbed sites in DNA by replication arrest and run-off replication. AB - We describe a technique for rapid fine mapping of sites of torsion-induced perturbations of DNA structure. The technique involves strand scission or chemical base modification at structurally perturbed sites, replication arrest in a double-strand DNA sequencing reaction, and size analysis of replication products by electrophoresis on sequencing gels. Besides being less complicated and faster than site identification by conventional end-labeling methods, the technique assures high sequence specificity through the use of oligomeric sequencing primers. This property should be useful for in vivo mapping of DNA structural perturbations with known sequence within complex genomes. PMID- 3040007 TI - Effects of low-amplitude pulsed magnetic fields on cellular ion transport. AB - Pulsed magnetic fields (PMFs) are widely used to treat difficult fractures of bone and other disorders of connective tissue. It is not clear how they interact with tissue metabolism, although it has been proposed that induced currents or electric fields impinging on cell membranes may modify their ion transport function. This hypothesis was tested by treating in vitro models for ion transport processes with short-term exposure to PMFs. No change occurred in active transport of potassium or calcium in human red cells or in calcium transport through an epithelial membrane. We considered less direct action on red cell membranes, that their permeability might be modified after PMF treatment, and also that PMFs might alter the extracellular ionic activity within connective tissue by interacting with its Donnan potential. Each of these studies proved negative, and we conclude that the PMF waveforms used here do not exert a general short-term effect on cellular ion transport. PMID- 3040008 TI - Temperature-specific inhibition of human red cell Na+/K+ ATPase by 2,450-MHz microwave radiation. AB - The ATPase activity in human red blood cell membranes was investigated in vitro as a function of temperature and exposure to 2,450-MHz continuous wave microwave radiation to confirm and extend a report of Na+ transport inhibition under certain conditions of temperature and exposure. Assays were conducted spectrophotometrically during microwave exposure with a custom-made spectrophotometer-waveguide apparatus. Temperature profiles of total ATPase and Ca+2 ATPase (ouabain-inhibited) activity between 17 and 31 degrees C were graphed as an Arrhenius plot. Each data set was fitted to two straight lines which intersect between 23 and 24 degrees C. The difference between the total and Ca+2 ATPase activities, which represented the Na+/K+ ATPase activity, was also plotted and treated similarly to yield an intersection near 25 degrees C. Exposure of membrane suspensions to electromagnetic radiation, at a dose rate of 6 W/kg and at five temperatures between 23 and 27 degrees C, resulted in an activity change only for the Na+/K+ ATPase at 25 degrees C. The activity decreased by approximately 35% compared to sham-irradiated samples. A possible explanation for the unusual temperature/microwave interaction is proposed. PMID- 3040009 TI - [Functionally important lysine residues in inorganic pyrophosphatase from E. coli. I. Interaction of inorganic pyrophosphatase with pyridoxal-5'-phosphate]. AB - Interaction of inorganic pyrophosphatase from E. coli with pyridoxal-5'-phosphate includes binding of the reagent at the active site through the phosphate group and then a reversible modification of one lysine residue in each of the enzyme's subunit. In the equilibrium state the protein's molecules contain both inactive modified and native subunits. A stable secondary amine is formed upon the sodium borohydride reduction of the modified protein. PMID- 3040010 TI - [Functionally important lysine residues in inorganic pyrophosphatase from E. coli. II. Isolation and characteristics of modified tryptic peptide and analysis of the functional role of lysine residues controlling enzyme activity]. AB - Inactivation of inorganic pyrophosphatase from E. coli by pyridoxal-5'-phosphate is due to the modification of a lysine residue located in the tryptic peptide with the Asp-Leu-Pro-Glu N-terminal sequence. In course of the enzymatic process this lysine-residue appears to be in the protonated state and either operators as the proton donor for the product of the enzymatic reaction or is involved in stabilization of the transition state. PMID- 3040011 TI - [A method of selective isolation of tryptophan- and cysteine-containing peptides by covalent chromatography. Analysis of the topography of the Na+,K+-ATPase alpha subunit]. AB - A procedure for highly selective isolation of tryptophan- and cysteine-containing peptides from protein hydrolysates has been developed on the basis of covalent chromatography. It includes incorporation of a thiol group into the tryptophan residues by sequential treatment of peptides with 2-nitrophenylsulfenyl chloride and beta-mercaptoethanol followed by immobilization on the corresponding supports via thiol-disulfide exchange. The technique is applicable to the analysis of the hydrolysate of the Na+, K+-ATPase alpha-subunit obtained by limited trypsinolysis of the membrane-bound enzyme. Fifteen tryptophan- and cysteine-containing tryptic peptides, which comprise the protein portions exposed outside the membrane, have been isolated in addition to those previously identified. This structural information allows unequivocal determination of boundaries of transmembrane segments of the alpha-subunit in the spatial model earlier proposed. PMID- 3040012 TI - [Study of the interaction between cytochrome c and cytochrome oxidase by tritium planigraphy]. AB - Treatment of molecular crystals of the bovine cytochrome oxidase and the cytochrome oxidase-cytochrome c complex with thermally activated tritium leads to highly labelled cytochrome oxidase preparations. HPLC separation of its subunits and measurements of radioactivity of each polypeptide allow to determine the shielding of cytochrome oxidase surface sites by cytochrome c in the complex. PMID- 3040014 TI - [Synthesis of phosphonate analogs of sphingomyelins and preparation of an affinity sorbent for sphingomyelinase purification]. AB - Phosphonate analogues of 2-N-stearoyl- (I) and 2-N-(undec-10-enoyl) sphingomyelins (II) have been synthesised. Compound (II) was used as a starting product for preparation of a sorbent for sphingomyelinase affinity chromatography. The double bond of the unsaturated undec-10-enoyl moiety of the phosphonate analogue (II) was oxidized, and the modified (II) was coupled to amino-Toyopearl HW-65 to give a sorbent containing 4 mumoles of ligand per milliliter of the swollen resin. PMID- 3040013 TI - [Chemical reactions in double-stranded nucleic acids. III. Synthesis of terminal inverted repeats of the IS1 element]. AB - Three 35 bp-DNA duplexes have been assembled from synthetic oligonucleotides by means of DNA ligase or 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide in two parallel series of experiments. The "top" strands of these duplexes correspond either to the imperfect (natural) or perfect terminal inverted repeats of the IS1. Tm of DNA duplexes composed of 2 to 6 different oligonucleotides were investigated by UV spectroscopy. It was shown that DNA ligase effectively joined oligonucleotides even under conditions of DNA duplex instability. However, there is a minimum duplex size (within the range of 9-15 bp) below which the enzymatic ligation is ineffective. Chemical assembly of duplexes took place only if the double helix was stable. The yield was 50% after two successive ligation cycles. Efficiency of the chemical ligation depends on the nature of the nucleotide units to be joined. PMID- 3040015 TI - [Molecular cloning of genes coding for tumor necrosis factor. Complete nucleotide sequence of the genome copy of TNF-alpha in mice]. AB - Using Sanger's technique and synthetic oligonucleotides derived from the known structure of cDNA, complete nucleotide sequence of the genomic copy of the mouse TNF-alpha gene has been determined. The length of the gene from transcription initiation site down to polyadenylation signal is about 2620 bp. Comparison with the previously described cDNA sequence revealed existence of 4 exons and 3 introns in the positions homologous to those of human TNF gene. The fourth exon codes for 88% amino acids of mature TNF-alpha. PMID- 3040016 TI - Hypertrophic osteoarthropathy, cutaneous vasculitis, and mixed-type cryoglobulinemia in a patient with nasopharyngeal carcinoma. AB - We describe a 27-year-old woman with a primary nasopharyngeal carcinoma who developed hypertrophic osteoarthropathy and cutaneous vasculitis. Her serum contained antibodies to Epstein-Barr virus and U1 RNP antigens. Cryoproteins isolated from her serum contained antibodies to U1 RNP and a protein with a molecular weight of 32 kd which reacted specifically with antibodies to U1 RNP. HLA typing revealed HLA-B7 and DR1; these have been reported to be increased in Japanese patients with rheumatic diseases who have autoantibodies to U1 RNP. These findings indicate that some features of the paraneoplastic syndrome in this patient might have been caused by immune complexes, part of which were formed by specific autoantibodies produced under genetically controlled conditions of immune responsiveness. PMID- 3040017 TI - The effect of dilazep on puromycin-induced rat renal mitochondrial dysfunction. AB - The effect of tetrahydro-1 H-1,4 (5H)-dipropanol bis(3,4,5 trimethoxybenzoate)hydrochloride monohydrate (dilazep, Comelian) on puromycin induced rat renal damage was investigated. In vivo study: Rats were divided into 3 groups, the control group; untreated, the puromycin group; puromycin (150 mg/kg) was injected intraperitoneally once, the dilazep + puromycin group; puromycin (150 mg/kg) was injected 1 h after intraperitoneal dilazep injection (2 mg/kg), and dilazep (2 mg/kg) was injected every 12 h until the end of the experiment. In each group, 84 h after puromycin injection, kidneys were isolated and renal mitochondria were prepared. The endogenous phospholipase activity in kidney homogenate was determined by high performance liquid chromatography. The activities of three segments (NADH-cytochrome c reductase, succinate-cytochrome c reductase and cytochrome c oxidase) of the electron-transport chain in mitochondria were measured enzymatically. In the puromycin group, phospholipase activity was increased and activities of all of three segments of the electron transport chain were decreased. In the dilazep + puromycin group, premedication with dilazep prevented activation of phospholipase and maintained mitochondrial electron-transport activity. In vitro study: Mitochondria prepared from intact rat kidney were incubated with phospholipase C. Activities of the mitochondrial electron-transport chain were deteriorated by phospholipase C. These results indicated that activation of endogenous phospholipase, which digests membrane phospholipids, essential components in maintaining mitochondrial electron transport activity, is responsible for the puromycin-induced renal damage. Premedication with dilazep prevented the damage by inhibition of the activation of phospholipase. PMID- 3040018 TI - In vivo effects of camostat mesilate on plasma kallikrein, plasma kininase II and renal kallikrein of man. AB - N,N-Dimethylcarbamoylmethyl-4-(4-guanidino-benzoyloxy)phenylacetat e methanesulfonate (camostat mesilate) is reported to be an effective inhibitor of plasma kallikrein. It was shown in vitro to inhibit not only plasma kallikrein, but also renal kallikrein and plasma kininase II. These inhibitory activities, however, were very weak. The inhibition of plasma kallikrein in human plasma was limited in time, since a rapid reactivation of plasma kallikrein was noticed when samples were incubated at room temperature. In order to establish whether camostat mesilate was able to inhibit plasma kallikrein, kininase II and renal kallikrein also in vivo the inhibitory activity of camostat mesilate on these enzymes was studied in 5 healthy volunteers. After an oral intake of a single dose of 600 mg of camostat mesilate, plasma kallikrein was inhibited significantly, while kininase II in plasma was unaffected. Renal kallikrein activity determined by urinary excretion of active kallikrein remained unchanged after camostat mesilate intake. Thus, the results demonstrate that camostat mesilate in vivo inhibits only plasma kallikrein and has no effect on the activity of kininase II or renal kallikrein. PMID- 3040019 TI - Trapidil derivatives as potential antiatherosclerotic drugs. AB - Trapidil, a triazolopyrimidine, and its derivatives are coronary vasodilating drugs. Trapidil reduces the serum level of low density lipoprotein- and very low density lipoprotein-cholesterol and increases the serum level of high density lipoprotein-cholesterol in hyperlipemic patients. The present study demonstrates that trapidil and five different trapidil derivatives inhibit the proliferation of cells cultured from grossly normal intima and fatty streaks of human aorta. The inhibiting effect of trapidil derivatives is about 60%, similar to the standard substance 3-isobutyl-1-methyl-xanthine (MIX). In cells cultured from atherosclerotic plaques trapidil and trapidil derivatives reduced the content of cholesteryl esters by 36% for trapidil and between 47% and 68% for 4 of 5 trapidil derivatives, respectively. The trapidil derivative AR 12463 (5 piperidino-7-[N-(n-amyl)-N-(beta-hydroxyethyl)amino]-s-triazolo[1,5- a]pyrimidine) reduces the free cholesterol content by 29%, but the other trapidil derivatives are without effect on this parameter. Four of five derivatives decrease the content of triglycerides by 53 to 70%. The synthesis of collagen is inhibited by the trapidil derivative AR 12463 (25%). Trapidil and other derivatives have a smaller or no effect on the synthesis of collagen. These effects of trapidil derivatives point to potential antiatherosclerotic properties. The possible mechanisms are discussed. PMID- 3040020 TI - [The effect of benzydamine on the generation and interaction of reactive oxygen species and cyclo- and lipoxygenase]. AB - Pharmacological mechanisms of benzydamine (Tantum) are studied which are of relevance for the antiinflammatory properties of this non-steroidal antiinflammatory drug (NSAID). Benzydamine most effectively inhibits the generation of reactive oxygen species by murine neutrophils (IC50 1.7 X 10(-5) mol/l). Piroxicam, indomethacin and acetylsalicylic acid are ineffective. Benzydamine, however, does not interfere with xanthine oxidase-dependent superoxide anion radical formation or epinephrine oxidation. The other tested NSAID are as well inactive. The findings confirm the missing cyclooxygenase inhibition of benzydamine (IC50 greater than 10(-3) mol/l), contrary to the other NSAID which are strong (indomethacin IC50 6 X 10(-8) mol/l; piroxicam IC50 2 X 10(-7) mol/l) or moderate cyclooxygenase inhibitors (acetylsalicylic acid IC50 10(-5) mol/l). LTB4 generation via the lipoxygenase is only inhibited by indomethacin (EC50 3.6 X 10(-5) mol/l). Benzydamine appears unique among other NSAID by its relatively strong interference with the generation of reactive oxygen radicals and the lack of cyclooxygenase inhibition. PMID- 3040022 TI - Cross-tolerance to metkephamid (LY127623) produced by morphine solution ingestion by mice. AB - The effects of morphine and metkephamid were determined on the writhing test in drug-naive mice and mice that had been exposed to morphine solutions as their sole fluid source for a 5- or 7-day period. The average dose of morphine consumed was 435 mg/kg, using either 0.5 mg/ml solution for 7 days or a 1 mg/ml solution for 5 days. The ingestion of these morphine solutions produced a marked tolerance to the analgesic effects of morphine and cross-tolerance to the analgesic effects of metkephamid. PMID- 3040021 TI - In vitro clearance of chylomicron triglycerides containing (omega-3) eicosapentaenoate. AB - Rat mesenteric lymph chylomicrons, containing triglycerides enriched with either [14C]oleic acid or [14C]eicosapentaenoic acid, were prepared by ultracentrifugation of lymph samples collected for 6 h after a single duodenal infusion of an emulsion containing 0.3 mmol of either fatty acid. After determination of protein and of total fatty acid content and composition, enriched chylomicrons were suspended in Krebs-bicarbonate buffer. Non-working hearts were perfused in a recirculating system for 45 min using the enriched chylomicron preparations. At 15 min intervals during perfusion, the media were assayed for total radioactivity, 14CO2 and 14C-labeled fatty acids associated with triglycerides, unesterified fatty acids, phospholipids, mono- and diglycerides. After perfusion, the hearts were extracted and assayed for total lipid radioactivity and isotope distribution among heart lipid fractions. With this membrane-supported lipoprotein lipase system, clearances of chylomicron triglycerides containing either fatty acid were identical, as were the myocardial uptakes of the fatty acids and oxidations to 14CO2. Furthermore, except for a significantly greater incorporation of eicosapentaenoate into myocardial phospholipids, tissue isotope distributions of the two labeled fatty acids were also the same. These studies suggest that at least the initial phases of peripheral clearance of chylomicrons enriched in omega-3 fatty acids is as efficient as with those containing oleate. PMID- 3040023 TI - Effects of restraint and naltrexone on the biphasic heart rate response to morphine in rats. AB - The effect of several doses of morphine (0, 0.5, 2, 5 or 10 mg/kg, i.v.) on heart rate was assessed in restrained and freely-moving rats. Morphine produced a dose dependent bradycardia followed by tachycardia. The magnitude and duration of bradycardia were greater in restrained rats, whereas the magnitude and duration of tachycardia were greater in unrestrained rats. Naltrexone (5 mg/kg) pretreatment completely blocked the biphasic heart rate response to morphine (8 mg/kg). When naltrexone was given after the bradycardic portion of the response, tachycardia declined to baseline levels. These results suggest that one or both components of the biphasic response are mediated by opioid receptors. PMID- 3040024 TI - Effect of long-term ethanol consumption on the rat ventricle. AB - The chronic ingestion of alcohol has been correlated with cardiac dysfunction. This study looked at the effect of chronic ethanol ingestion on the rat ventricle. The studies were carried out on hearts from male Long-Evans hooded rats, pair-fed on ethanol (E) or normal (N) liquid diet. The E rats received 35 39% of calories as ethanol. The studies were carried out after 40 weeks on the diet. The data show the E rats had reduced papillary muscle function, and increased incidence of isoproterenol induced extra beats and failure. There was no difference in responsiveness to isoproterenol, alpha, beta, or muscarinic receptor number or agonist binding characteristics between hearts from E and N rats. The cardiac dysfunction in the E group is thought to be due to possible membrane structural changes, or to changes in the characteristics of the autonomic receptor system beyond the receptor level. PMID- 3040025 TI - Pharmacological profile of delta 9-THC carbamate. AB - 1-N,N-bis-(Dichloroethyl)carbamate-delta 9-THC (THC carbamate), a nitrogen mustard analog of delta 9-THC, was recently synthesized as a potential anti-tumor agent. The decrease in spontaneous activity, induction of hypothermia, and the antinociceptive properties of THC carbamate and delta 9-THC were compared. THC carbamate and delta 9-THC were administered by a number of peripheral routes as well as intraventricularly (ivt). THC carbamate lacked cannabinoid activity following peripheral administration, with the exception of iv administration which produced very weak cannabimimetic effects. In contrast, THC carbamate was equipotent to delta 9-THC in reducing rectal temperature by 3 degrees C, and 5 times less active in decreasing spontaneous activity following ivt administration. The apparent lack of central effects following peripheral administration might limit the effectiveness of THC carbamate as an anti-emetic agent, but its use as a site-directed alkylator (a receptor probe) holds promise. PMID- 3040026 TI - Production of monoclonal antibodies using spleen cells from nude mice bearing human tumors. AB - Monoclonal antibodies were generated with spleen cells from nude mice bearing human tumors. Three grafted tumors were selected because of their difference in metastatic ability in nude mice. Two were non-metastasizing carcinomas and one a highly metastasizing adenocarcinoma of the lung (Bur-tumor). Two normal nude mice were used as controls. Culture supernatants were screened by immunoperoxidase using frozen sections from both the immunizing human tumor and normal human tonsil to detect unexpected monoclonal antibodies. Two kinds of monoclonal antibodies were obtained. The first were directed against miscellaneous membrane and/or cytoplasmic antigens expressed by normal cells (e.g. normal tonsillar epithelium). Most of these antibodies corresponded to auto-antibodies and their frequency did not appear to be influenced by the fact that the mice were with or without grafted human tumors. The second type of antibodies were directed against tumor-associated antigen and generated only by fusing splenocytes from nude mice bearing the metastasizing lung adenocarcinoma. On frozen sections Bur-1-anti tumor antibody stained all but one lung carcinoma. Occasional carcinomas originating from other organs such as pancreas and breast were also labelled. On the other hand, more than half of the cases of so-called "malignant histiocytosis" (Ki-1 lymphoma) seemed to express this epithelial antigen. Some normal cells in the lung, pancreas (acini), and kidney (distal tubules) also bound Bur-1 antibody. These results suggested that Bur-1 antibody could be related to some antibodies already described, directed against epithelial membrane antigens. When Bur-tumor was analysed by immunoblotting with Bur-1 antibody a positive reaction was obtained with material migrating in the kD-45kD molecular-weight region. PMID- 3040027 TI - [Glandulo-cystic polyps of the stomach. Apropos of 12 cases and review of the literature]. AB - We report 12 cases of cystic glandular fundic polyps detected in 7 women and 5 men. As in the 238 similar previously reported cases these lesions were always located in the fundic gastric mucosae and did not involve the muscularis mucosae. They were discovered by endoscopy in patients with minor abdominal discomfort. They were less than 5 mm in diameter and were more often multiple and sessile; only a few of them were pedunculate. They consisted of mucosal cysts lined with cuboidal, parietal or chief cells and surrounded with a normal lamina propria. If in 213 cases, these polyps were not associated with polyposis coli in 37 cases they occurred with a Gardner's syndrome or a familial adenomatosis coli. These benign lesions are without malignant potential. Careful gastroscopic follow up with biopsy is recommended for all patients with multiple gastric polyps. The etio-pathogenesis of these polyps is still unknown. Their relationship to familial polyposis coli or to Gardner's syndrome is obscure. PMID- 3040028 TI - [Malakoplakia at 3 sites: the bladder, urethra and kidney. Isolation and ultrastructural study of blood monocytes]. AB - We studied the blood mononuclear cells in a seventy-four-year old man who had urinary tract malacoplakia located to bladder, ureter and kidney. The blood mononuclear cells were isolated as described by Boyum [2] and studied by electron microscopy. They did not show bacilliform bodies or bacteria in the phagolysosomes. The microfilaments and the microtubules were not easily identifiable in the mononuclear cells of the patient. In the control, the internal skeleton of the mononuclear cells was normal. This ultrastructural finding may suggest that there is a relation between microfilaments and microtubules lesion and the low level of cyclic G.M.P. described by Abdou et al. PMID- 3040029 TI - Carcinomatous neuropathy. An ultrastructural study of ten cases. AB - Superficial peroneal nerve biopsies were studied from 10 patients with carcinomatous neuropathy. There were 6 patients with the sensorimotor form and 4 patients with the sensory form. In both forms of neuropathy, histometric studies showed a variable loss of myelinated fibers which was generally most severe in the sensory form. In most of the cases of the sensorimotor form, ultrastructural studies showed segmental demyelination with onion bulb formation and regeneration of myelinated fibers. In the sensory form, there was no segmental demyelination, the regeneration of myelinated fibers was minimal and all the cases showed a loss of unmyelinated fibers. In sensory neuropathy, the histometric and ultrastructural changes reflect the massive acute destruction of ganglion cells in the dorsal root ganglia. In sensorimotor neuropathy, the alterations indicate a slowly progressive degenerating process in which both axonal degeneration and segmental demyelination occurred. PMID- 3040030 TI - Interleukin 2 and lymphokine-activated killer cells in the treatment of childhood primary hepatocellular carcinoma--a preliminary report. AB - Recombinant interleukin-2 (RIL-2) and lymphokine-activated killer (LAK) cells were administered to 2 boys with the end-stage of primary hepatocellular carcinoma (HCC); the efficacy and toxicity were evaluated. Immunologically, the natural killer and LAK activities were enhanced. Clinically, the side effects were similar to those reported for adults but milder. This kind of treatment may be considered for children with the early stages of hepatocellular carcinoma. PMID- 3040031 TI - Mammary carcinoma. Comparison of nuclear DNA content from in situ and infiltrative components. AB - Twenty-eight mammary carcinomas were analyzed with respect to their nuclear DNA content. Ten of the carcinomas were entirely in situ (noninfiltrative) while 18 showed areas of both infiltrative and noninfiltrative growth. The DNA content of individual tumor cells was measured in sections from the original paraffin embedded specimens. In the tumors that had noninfiltrative as well as infiltrative zones, DNA analyses were performed in both areas. Comparison between the DNA patterns obtained from these different areas of the same tumor showed very close agreement. Both groups of tumors (those with and those without areas of invasion) contained some cases that showed a euploid DNA pattern and some cases that showed an aneuploid pattern. Furthermore, analysis of the DNA content of regional lymph node metastases in seven of the invasive cases did not show an increased aneuploidy in the metastases. The results suggest that, in mammary carcinomas, invasive and noninvasive tumors cannot be distinguished by DNA analysis and that tumor progression does not seem to be associated with a significant alteration of the nuclear DNA content. PMID- 3040032 TI - Effects of lateral hypothalamic lesions on food intake of rats during exposure to cold. AB - Lateral hypothalamic damage impaired both physiological and behavioral responses of rats during exposure to a 5 degree C environment. The brain-damaged animals usually did not conserve heat in the cold as well as control rats did, nor did they always increase their caloric intake to meet their energy needs. However, when given sucrose solution to drink instead of water, they did increase their ingestion of chow in response to cold exposure. It is likely that the elevated consumption of palatable fluid served to relieve dehydration and thereby removed its constraints on eating, thus permitting hyperphagia to occur. In contrast to these results, rats with large dopamine-depleting brain lesions, produced by intraventricular 6-hydroxydopamine treatments, always increased food intake when exposed to cold stress and demonstrated no apparent problems in peripheral vasoconstriction. Thus, it is unlikely that striatal dopamine depletions account for either the impaired feeding response or the inadequate heat conservation of rats with lateral hypothalamic lesions during cold stress. PMID- 3040033 TI - Thalamocortical relations in taste aversion learning: II. Involvement of the medial ventrobasal thalamic complex in taste aversion learning. AB - Previous neurobehavioral studies have implicated the gustatory thalamocortical relay as a functional substrate of conditioned taste aversion (CTA) learning. These experiments were conducted to examine the involvement of the gustatory thalamic nuclei in fundamental taste reactivity, gastrointestinal reactivity, and CTA learning. In Experiment 1, bilateral electrolytic lesions were produced in the medial ventrobasal thalamic complex (VBm), including the thalamic gustatory nuclei, in one group of rats. A separate group of rats received control lesion placements in the mediodorsal-periventricular (MD-PV) thalamic nuclei. Animals then received preference-aversion taste tests followed by CTA conditioning. At the conclusion of conditioning, lesions were produced in the anterior insular gustatory neocortex (AIGN) to evaluate whether or not the AIGN contributed to CTA learning in animals lacking VBm thalamus. Results of Experiment 1 indicated that control lesions did not disrupt taste reactivity, gastrointestinal reactivity, or CTA learning. Destruction of VBm thalamus attenuated taste reactivity to sucrose, citric acid, and quinine hydrochloride; however, such lesions did not impair normal taste reactivity to sodium chloride. Lesion placements in VBm thalamus also did not reliably impair gastrointestinal reactivity to ingested LiCl. Elimination of VBm thalamus markedly attenuated CTA learning. Results of neocortical lesion manipulations showed that the AIGN contributed to initial CTA learning in animals lacking MD-PV thalamus but that the AIGN did not mediate initial CTA learning in animals lacking VBm thalamus. Whether animals lacking VBm thalamus used olfactory cues associated with drinking solutions to acquire CTAs was evaluated in Experiment 2. Results of Experiment 2 demonstrated that animals lacking VBm thalamus and the olfactory bulbs could not acquire aversions to ingested LiCl following eight conditioning trials. These experiments demonstrate that destruction of VBm thalamus, including the gustatory thalamic nuclei, is sufficient to prevent CTA learning. PMID- 3040034 TI - The rat fetus in its environment: behavioral adjustments to novel, familiar, aversive, and conditioned stimuli presented in utero. AB - With the pregnant rat under ether anesthesia, rat fetuses were exposed on Day 17 of gestation to a taste/odor stimulus (mint) injected into the amniotic fluid and/or lithium chloride injected into the peritoneum. Behavior of injected fetuses was directly observed on Day 19 of gestation following chemomyelotomy and laparotomy of the female and immersion of the uterus into a warm saline bath. With these procedures, a series of four experiments was conducted to assess the behavioral effects of (a) the mint taste/odor alone, (b) the LiCl alone, (c) the pairing of mint and LiCl on the day of conditioning, and (d) the reexposure to mint after an earlier pairing of mint and LiCl. These experiments provide clear evidence that rat fetuses are capable of forming conditioned taste/odor aversions as early as Day 17 of gestation and, further, that rat fetuses are capable of expressing these learned aversions in utero. PMID- 3040035 TI - Administration of dexamethasone prior to training blocks ACTH-induced recovery of an extinguished avoidance response. AB - Pretest administration of ACTH has been shown to produce recovery of an extinguished avoidance response. Presumably this effect is found because endogenous ACTH is a component of the original training memory. However, another possible explanation of this finding is that administration of the peptide acts as a novel stimulus that "disinhibits" the extinguished response. In order to test this "disinhibitory" hypothesis of ACTH-induced recovery of an extinguished avoidance response, some subjects were given dexamethasone 2 hr prior to training and extinction. This synthetic glucocorticoid is effective in blocking endogenous release of ACTH. Thus, ACTH should not be a component of the training memory in subjects given dexamethasone prior to training and extinction but would be a relatively novel stimulus condition at testing. Pretest administration of ACTH was found to be effective in alleviating performance deficits induced by extinction only for subjects given saline prior to training and extinction. Administration of ACTH had no effect on the avoidance responding of subjects given dexamethasone. These findings suggest that pretest administration of ACTH affects retrieval processes rather than acts as a disinhibitor. PMID- 3040036 TI - ATP-dependent proteolysis and the role of ubiquitin in rabbit cardiac muscle. AB - A soluble ATP/Mg2-dependent proteolytic system from rabbit cardiac muscle has been identified (m ca. 310 kDa) and purified ca. 9-fold. This enzyme which splits the substrate [3H]globin and 125I-bovine serum albumin (125I-BSA) has many similarities to the ATP-dependent proteolytic enzyme system from reticulocytes which utilizes ubiquitin: 1) The specific activities in reticulocyte lysates and cardiac muscle extracts are of the same magnitude (0.5-1 arb. unit/mg). 2) The binding and elution behavior on DEAE-cellulose is similar. 3) In both cases the pH optimum (substrate 125I-BSA) is pH 7.6. 4) Both enzymes are inhibited by hemin, NEM and iodoacetate but not e.g. by leupeptin, or inhibitors of serine proteases. 5) Neither enzyme system can utilize ATP-analogs such as AMP-CPP, AMP PCP, AMP-PNP or ATP-gamma-S. There are however also significant differences: 1) The enzyme system from cardiac muscle is fully active in the absence of ubiquitin and cannot be activated by this peptide. 2) The enzyme from cardiac muscle can degrade methylated BSA. 3) The cardiac muscle enzyme can be further purified on Sepharose 4B; the enzyme from reticulocytes is inactivated by this procedure. 4) The cardiac enzyme cannot be inactivated by ribonuclease as the reticulocyte counterpart. Although ubiquitin does not appear to play a role in the isolated ATP/Mg2-dependent proteolytic system from cardiac muscle, it is demonstrated for the first time that 125I-ubiquitin can be conjugated to a wide variety of cardiac muscle proteins in vitro in an ATP-dependent manner. Apparent molecular masses of major conjugates were: 185 kDa, 140 kDa, 85 kDa, 65 kDa, 46 kDa, 38 kDa and 36 kDa as estimated by discontinuous SDS gel electrophoresis. Addition of purified phosphorylase kinase to cardiac muscle extract changed the ubiquitination pattern by the appearance of two novel protein bands. It is concluded that the ATP/Mg2 dependent proteolytic system of cardiac muscle must be differentiated from the proteolytic system of reticulocytes mainly because of its ubiquitin-independence. Nevertheless the conjugation of 125I-ubiquitin to many muscle proteins is a strong indication for a crucial role of this interesting peptide in striated muscle. PMID- 3040037 TI - Studies on immunoassays of peptide factors. III. Gastrin/iso-1-cytochrome C as immunogen for raising anti-gastrin antisera. AB - Iso-1-cytochrome c contains in penultimate position of its sequence a cysteine residue which in analogy to the known tertiary structures of cytochromes c should be exposed on the surface of the protein. Its selective reaction with N alpha maleoyl-beta-alanyl-human-little-gastrin-I-[2-17] led to a well characterized and homogeneous gastrin conjugate to be used as immunogen in rabbits. The antisera raised by this procedure exhibited a degree of specificity for the hormone gastrin parallel to that of the gastrin receptor. This is clearly documented by comparison of the immune crossreactivities of gastrin-peptides of increasing chain length and of fragments corresponding to various regions of the hormone molecule with their biological activity. The immune response provoked in the animals by the use of an homogeneous immunogen was found to be highly reproducible in terms of specificity of the antigastrin antibodies. PMID- 3040038 TI - Studies on immunoassays of peptide factors. V. Synthesis of cholecystokinin-58-[1 11]/iso-1-cytochrome c conjugate. AB - In previous studies on model compounds we have found that the maleimide function is sufficiently stable under the conditions of peptide synthesis to allow its incorporation at preselected positions of a peptide chain in earlier steps of the synthetic route. Taking advantage of this observation the N-terminal undecapeptide of canine cholecystokinin-58 containing at its N-terminus the maleimide group became accessible in high yields as chromatographically homogenous and analytically well characterized compound. Via the incorporated anchor group the undecapeptide was linked selectively at its N-terminus to the cysteine residue 107 of iso-1-cytochrome c to yield a well characterized conjugate of 1:1 stoichiometry for immunization experiments. PMID- 3040039 TI - Analysis of density changes and chemotactic receptors of leukocytes from chronic hemodialysis and peritoneal dialysis patients. AB - Analysis of standard Ficoll-Hypaque (density = 1.077 g/ml) separation profiles of peripheral white blood cells (WBC) from patients undergoing hemodialysis (HD) demonstrated that dialysis caused a marked decrease in the density of polymorphonuclear leukocytes (PMN) resulting in about 50% of these cells separating with the mononuclear cells. In vitro exposure of normal control peripheral blood to HD membranes as well as to the purified chemotactic factors C5a, C5ades-Arg, and formyl-Met-Leu-Phe (fMLP) also resulted in PMN density changes which altered the Ficoll-Hypaque separation profiles of WBC. Therefore, these results imply that C5a generation, resulting from complement activation by the HD membrane, induced the density changes in the PMN from HD patients. Further studies using flow cytometry and fluorescein-labeled chemotactic factors (C5a, formyl-Met-Leu-Phe-Lys [fMLPL] and casein) indicated that HD patients had a significant reduction in the ability of their PMN and monocytes to bind C5a. This contrasted with the findings of no significant difference in the percentage or fluorescence intensity of HD patients' PMN or monocytes binding casein or fMLPL. Functional studies to analyze chemotactic-factor-mediated responses indicated that there was a decreased ability of HD patients' PMN and monocytes to generate superoxide anion, produce H2O2 and release myeloperoxidase in response to both C5a and fMLP. Additional studies evaluated the binding of chemotactic factors to PMN and monocytes from normal blood following passage through a hemodialyzer and from patients undergoing HD. Analysis of receptor binding by control cells passed through the dialyzer showed that there was a progressive decrease in the percentage of C5a-receptor-positive PMN and monocytes but no change with casein or fMLPL. In contrast, peripheral PMN and monocytes from chronic renal failure patients on HD showed no difference in C5a, casein or fMLPL receptors during the course of HD as compared to the predialysis period. This appears to be attributable to a difference in the regulation of the C5a that is generated as a result of the dialysis-membrane-induced activation of the complement system. Although C5a has been shown to be continuously generated during the course of HD, these patients show no modulation of their C5a receptors during the course of HD or when their whole blood is exposed to dialysis membrane fibers. These findings suggest that there are mechanisms functioning in chronically dialyzed patients to protect them from the effects of excessive C5a generation during HD.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3040040 TI - Monoclonal-oligoclonal immunoglobulins in HTLV III infected subjects: HTLV III induced or HTLV III-associated? AB - Sera from 55 patients with positivity for anti-HTLV III antibodies, including 24 "healthy" subjects, 29 LAS and 2 AIDS, were examined by means of high resolution agarose gel electrophoresis and immunofixation, in order to evaluate the occurrence of monoclonal-oligoclonal immunoglobulin patterns. Monoclonal oligoclonal immunoglobulins were found in 20.8% of "healthy" subjects and in 72.4% of LAS patients. Furthermore, a lower but significant prevalence of monoclonal-oligoclonal immunoglobulins was found in a control group including patients with high-titre positivity for anti-CMV and anti-EBV antibodies. Considering the consistent association between EBV-CMV and HTLV III infection, it is concluded that at least a part of monoclonal-oligoclonal immunoglobulins found in LAS-AIDS patients are likely to be induced by these associated viral infections. On the other hand, the finding of monoclonal-oligoclonal immunoglobulins in "healthy" HTLV III-positive subjects points out a possible direct stimulatory effect of HTLV III on B-cell lineage. PMID- 3040041 TI - In situ hybridization of papillomavirus DNA in head and neck squamous cell carcinomas. AB - Cytospin preparations of fine-needle aspirates obtained from five squamous cell carcinomas of the head and neck (four metastatic in lymph nodes and one primary) were tested for human papillomavirus (HPV) genotypes 6, 11, 16, and 18 by in situ hybridization using biotinylated DNA probes. All five carcinomas showed hybridization with HPV-11, four strongly reactive, and one also weakly hybridized with HPV-16. Genotypes 6 and 18 showed no evidence of hybridization. PMID- 3040042 TI - Bilateral synchronous mixed tumors of the parotid glands. AB - Bilateral synchronous mixed tumors of the parotid glands are uncommon. In this case, they occurred in association with multiple benign tumors elsewhere in the body. The larger of the two parotid neoplasms was asymptomatic and undetected on physical examination, even after its presence had been established by computed tomographic scan. PMID- 3040043 TI - Ion transport mechanisms in renal tubular cell membranes. PMID- 3040044 TI - ACTH, beta-endorphin, beta-lipotropin and cortisol response to an agonistic athletic performance is related to the energetic characteristics and to the duration of the competition. PMID- 3040045 TI - [Changes in the plasma levels of ACTH and cortisol during the inhalation of nicotine from cigarette smoke in normal smoking and non-smoking subjects]. PMID- 3040046 TI - [Various hormonal parameters (ACTH, cortisol, somatotropic hormone and prolactin) following administration of a single high dose of pantethine in healthy subjects]. PMID- 3040047 TI - [Long-term outcome of shoemaker's polyneuropathy]. PMID- 3040048 TI - [Effect of serotonin on the cytochrome oxidase activity and on blood glucose in normal and pregnant rats (20 C in the daytime)]. PMID- 3040050 TI - Papillomaviruses in human cancer. AB - Twelve types of human papillomaviruses (HPV) have been isolated thus far from papillomatous and Bowenoid lesions of the human genital tract. Four type, HPV 6, 11, 16 and 18 are most frequently found. HPV 6 and 11 cause the typical genital warts (condylomata acuminata) and mild dysplastic lesions of the cervix characterized by a high degree of koilocytotic atypia. HPV 16 and 18 are preferentially found in Bowenoid papulosis and Bowen's disease at external genital sites. Moderate and severe cervical dysplasias with little or no koilocytosis appear to be common manifestations of these infections at cervical sites. HPV 16 is found in approximately 50% of all cervical, penile and vulvar cancers, HPV 18 in close to 20%. The majority of the remaining tumors reveals evidence for infections with additional types of HPV. Several cell lines have been identified containing either HPV 18 or HPV 16 genomes. The state of viral DNA in Bowenoid precursor lesions differs from that of cervical carcinomas. The former contain episomal DNA whereas integration seems to be a regular event in carcinomas. Integration regularly affects the E1-E2 open reading frames of HPV 16 or HPV 18 DNA. Fusion transcripts from the integrated HPV DNA (E6-E7 region) and adjacent host cell DNA have been documented. The available data support a causative role of specific HPV infections in the etiology of human genital cancer. PMID- 3040049 TI - Molecular characterization of a conjugative R-lac plasmid in Salmonella typhi isolated from a patient with typhoid fever. AB - In a strain of Salmonella typhi isolated from a patient with post-operative typhoid fever, a 217-Kb conjugative plasmid has been detected which codes for lactose fermentation and drug resistance to sulphonamides (SA), chloramphenicol (CP), ampicillin (AP), kanamycin (KM) and trimethoprim (TP). The physical and genetic map was constructed. The lac region containing the lacI gene, the regulatory region and the lacZ and lacY genes was shown to be structurally identical to that of Escherichia coli K12. A transposon carrying genes for resistance to CP and AP, as well as four copies of IS1-like elements, were detected and localized on the plasmid. Their role in the emergence of such a large plasmid is discussed. PMID- 3040051 TI - Current concepts of intraepithelial neoplasia in the uterine cervix (CIN). AB - The unifying concept of precancerous events in the epithelia of the uterine cervix is reviewed in the light of epidemiologic and morphologic observations. The role of human papillomavirus (HPV) infection as a factor in the genesis of precancerous states and carcinoma of the uterine cervix is assessed. Although cytologic, histologic, ultrastructural and molecular biologic data support the concept of HPV as an oncogenic virus, the precise mechanisms linking the infection with cancer are not known at this time. A hypothesis of the presumed sequence of events is presented. PMID- 3040052 TI - Genetic analysis of a new subgroup of human and simian T-lymphotropic retroviruses: HTLV-IV, LAV-2, SBL-6669, and STLV-IIIAGM. AB - A new primate retrovirus, STLV-IIIAGM, has been recently isolated from healthy African green monkeys and is apparently nonpathogenic in its natural host. However, spontaneous infection as well as inoculation of STLV-IIIAGM into macaques induces a disease with clinical features that resemble human AIDS. Independent isolates of human retroviruses, serologically closely related to STLV IIIAGM, have been obtained from healthy individuals (HTLV-IV) and patients with immunodeficiency (LAV-2FG and SBL 6669) from West Africa. The latter have also been referred to as HIV-2 because, like HTLV-III/HIV-1, they may be associated with immune deficiency, or as West African human retroviruses because of their prevalence and probable origin from that region. We have molecularly cloned the STLV-IIIAGM genome and have generated probes from the gag-pol and envelope genes to analyze the genetic relatedness of these simian and human retroviruses. Our results indicate that all these retroviruses are genetically closely related to each other, HTLV-IV and STLV-IIIAGM differing only by a few restriction enzyme sites while LAV-2FG and SBL 6669 exhibit greater polymorphism from HTLV-IV/STLV IIIAGM. These data mirror the variable degree of relatedness among members of the first subgroup of human retroviruses, HTLV-III/HIV. PMID- 3040053 TI - A new human retrovirus isolate of West African origin (SBL-6669) and its relationship to HTLV-IV, LAV-II, and HTLV-IIIB. AB - A new human retrovirus of West African origin (SBL-6669) has been isolated from a patient with immunological and clinical signs of immunodeficiency. Using radioimmunoprecipitation assays (RIPA) and Western blot (WB) tests with human sera, the new virus isolate has been compared with HTLV-IV, LAV-II, and the HTLV IIIB prototype strain of the human immunodeficiency virus (HIV). The West African isolates appeared to be members of the same virus group since their glycoproteins were antigenically indistinguishable. West African sera showed no detectable cross reaction with HTLV-IIIB glycoproteins. The external glycoprotein in the different virus strains only showed minor variations in size. The size of the transmembranous protein was not unambiguously defined. In the West African virus isolates a 30-35 kD protein was seen similar to the protein previously described possibly to represent this component. However, in SBL-6669 a distinct 41 kD protein was also identified. There were interstrain variations in the size of several viral proteins among the West African virus isolates. Only minor differences were seen between SBL-6669 and LAV-II. The variations were most pronounced in two core proteins corresponding to the 19 kD and 24 kD proteins of HTLV-IIIB. In addition, West African human retroviruses appear to differ in pathogenicity. LAV-II and SBL-6669 are associated with immunodeficiency, whereas HTLV-IV was isolated from healthy individuals. Since further spread of these viruses to other parts of the world is imminent, it is necessary to consider their antigenic and immunogenic properties in serodiagnosis of HIV infections and in planning for immunoprophylactic interventions. PMID- 3040055 TI - Complete nucleotide sequences of functional clones of the AIDS virus. AB - To examine the mechanism of lymphocytotoxicity induced by human T-lymphotropic virus type III/lymphadenopathy associated virus (HTLV-III/LAV), an in vitro model has been developed. Introduction of an HTLV-III/LAV proviral clone, HXB2, into normal lymphocytes results in the production of virions and cell death. The complete nucleotide sequence of the proviral form of HXB2 has now been determined. Its structure is quite similar to that previously determined for HTLV III/LAV clones whose biological capacities had not previously been demonstrated. The biological function of two additional clones of HTLV-III/LAV, BH10 and HXB3, are reported. Clone BH10 which lacks the 5' long terminal repeat sequences (LTR) and a portion of the 3' LTR is reconstituted by substituting the corresponding sequences of HXB2 and is shown to be capable of generating infectious cytopathic virions. Clone HXB3, which has been partially sequenced, is also found to be capable of producing lymphocytopathic virus. Clone HXB3 differs from HXB2 in its lack of a termination codon in 3' orf, demonstrating that 3' orf plays no major role in virus replication or cytopathic activity. These data provide the necessary background to allow the identification of viral determinants of replication, cytopathic activity, and antigenicity using these functional proviral clones. PMID- 3040054 TI - Transcription directed by the HIV long terminal repeat in vitro. AB - The long terminal repeat (LTR) of the AIDS virus (HIV) has been found to contain promoter sequences that are active in uninfected HeLa whole cell and nuclear extracts. Here we report that elements upstream of position -104 (start site +1) do not affect transcriptional activity in vitro whereas sequences between -104 and -57 are required for such activity. Using a reconstituted RNA polymerase II system, we demonstrate that a partially purified fraction containing Spl not only stimulates, as was previously reported, but is required for accurate initiation of transcription directed by the HIV LTR. In addition, based on a computerized analysis, we report the presence of a region in the HIV LTR (positions -151 to 80) that is similar to the 72 base pair enhancer element of SV40 and that includes a highly conserved segment also present in the cytomegalovirus enhancer. Moreover, the HIV and HTLV-I LTRs are shown to share a region of similarity that includes the 21 base pair motif found in the enhancers of the human and bovine T lymphotropic viruses. The R region of the HIV LTR is found to have two extensive regions of dyad symmetry rather than one as was previously reported. The significance of these observations for HIV pathogenesis is discussed. PMID- 3040056 TI - [Alkaline phosphatase activity in human gliomas as revealed by light and electron microscopy]. AB - Precise localization of alkaline phosphatase (ALPase) activity in human gliomas was examined by light and electron microscopy in association with malignant transformation, paying much attention to changes in blood-brain barrier. Materials used were eight cases of gliomas four of which were astrocytoma grade 2, one astrocytoma grade 3, and three, glioblastoma multiforme, respectively. Specimens were quickly fixed in cacodylate-buffered 2% glutaraldehyde at 4 degrees C for 1 hour and rinsed overnight in the same buffer. Frozen or nonfrozen sections (40 microns thick) were made and incubated at 20 degrees C for 40 min, and processed for light and electron microscopy. For the demonstration of ALPase activity, the lead citrate method (Mayahara et al., 1967) was employed. By light microscopy, ALPase activity appeared to be mainly restricted to the capillary wall. By electron microscopy, reaction product representing ALPase activity was distributed in the plasma membrane of endothelial cells both in astrocytomas and in glioblastoma. In astrocytoma grade 2, activity was primarily localized along the abluminal surface of endothelial cells. In glioblastoma, on the other hand, ALPase activity was positive on the luminal surface of the plasma membrane of endothelial cells. It was much more intense than that along the abluminal surface. Regional differences in enzyme cytochemistry may represent functional heterogeneity in the endothelial cell membrane. In brain tumors, changes in distribution pattern of enzyme activity were visualized in the present study in association with glioma malignancy. This might represent a functional aspect of changes in blood-brain barrier in human glioma tissue during course of its malignant transformation. PMID- 3040058 TI - Platelet alpha 2-adrenoceptor binding does not predict brain alpha 2-adrenoceptor function. PMID- 3040057 TI - Dentine hypersensitivity: a comparison of five toothpastes used during a 6-week treatment period. PMID- 3040059 TI - Amiodarone and multilamellar inclusion bodies. PMID- 3040060 TI - Cyclosporin A and its analogues as modifiers of adriamycin and vincristine resistance in a multi-drug resistant human lung cancer cell line. PMID- 3040062 TI - Atypical manifestations of pulmonary adenoid cystic carcinoma. AB - Pulmonary adenoid cystic carcinoma (PACC) typically arises in large airways. A patient who presented with a peripheral lung mass due to PACC is reported. She was found to have multiple pulmonary nodules due to PACC 11 years after resection of the original tumour. We emphasize that 10-15% of patients with PACC present with peripheral tumours. PMID- 3040061 TI - Classification of testicular cancer in incidence and mortality statistics. PMID- 3040063 TI - Connective tissue responses to some heavy metals. II. Lead: histology and ultrastructure. AB - Lead loaded ion exchange resin beads implanted into the loose connective tissue of the rat pinna induced local lesions which differed widely from those of the control (sodium loaded) beads (Ellender & Ham 1987). These lesions were characterized by changes in the granulation tissue and the approximating connective tissue. Granulation tissue contained mononuclear phagocytes in various guises, and some cells with intranuclear inclusion bodies. The matrix of the granulation tissue contained collagen fibrils having a wide range of diameters suggestive of altered collagen biosynthesis. Foci of collagen mineralization occurred in zones of combined trauma and lead impregnation. Once mineralized they became enveloped by giant cells and epithelioid cells. Lead in damaged tissues is thought to modify the protective mechanism of calcification inhibition and the biosynthesis of the matrix. PMID- 3040064 TI - Lysosomal enzyme activity and fibroblast stimulation of lavage from guinea pigs exposed to silica dust. AB - Guinea pigs were exposed to silica dust for 3 weeks and examined up to 1 year thereafter. The activity of several lysosomal enzymes in lung lavage fluid (LLF) increased from 8 weeks after cessation of exposure whereas the level in aveolar macrophages decreased. Collagen synthesis in fibroblast cultures exposed to LLF was also increased except at 24 weeks after exposure. The results indicate that 8 weeks and later after cessation of exposure to silica dust there are cellular changes in the lung which can be related to the exposure, although typical silicotic lesions were not observed till 1 year after the exposure. PMID- 3040065 TI - Hexokinase and adenylate kinase activities in aorta, heart muscle and skeletal muscle from uraemic rats. AB - The effect of parathyroidectomy and/or vitamin D on the development of arterial and myocardial lesions was studied in rats with moderate uraemia. The activities of hexokinase and adenylate kinase in the aorta, myocardium and skeletal muscle were measured and the incidence of aortic calcification and muscle cell necrosis determined. There was a decreased hexokinase activity in the aorta, myocardium and skeletal muscle from uraemic rats. Adenylate kinase showed an increased activity in the same tissues. Parathyroidectomy as well as I-alpha hydroxycholecalciferol in a dose of 3 ng/100 g b.w. normalized these activities to a great extent. This effect did not occur when 10 ng/100 g b.w. was given. Parathyroidectomy in combination with a low dose of I-alpha-OH-D3 reduced the incidence of myocardial necrosis. Aortic calcifications were found in uraemic animals given 10 ng/100 g b.w. of I-alpha-hydroxycholecalciferol. In this group increased activity of adenylate kinase was found in calcified aortae but not in non-calcified aortae. The study shows that uraemia causes metabolic changes in the aorta, myocardium, and skeletal muscle which may partly be prevented by parathyroidectomy and by low doses of vitamin D. It also indicates some parallelism between these metabolic changes and the development of histologically demonstrable lesions in the aorta. PMID- 3040066 TI - Cellular basis of host defence in pyelonephritis. III. Deletion of individual components. AB - Hosts were depleted of individual cellular components to determine the effects of these manipulations on cellular defence mechanisms in acute and chronic pyelonephritis. T-lymphocytes were found to have little or no involvement in host protection but cyclosporin A administration had a dramatic effect on the gross pathology and bacteriological status of experimentally induced pyelonephritis. This change represented a major depression of host defence status. Cyclosporin A also activated resolved lesions in chronic pyelonephritis, associated with an increase in bacterial numbers. Administration of antineutrophil serum also led to a 1000-fold increase in bacterial numbers in the acute phase but had little effect on the host-parasite balance in chronic pyelonephritis. Macrophage blockade, on the other hand, did not affect the course of either acute or chronic infection. These studies have provided additional information on the immunobiology of experimental pyelonephritis and have focussed attention on the role of neutrophils, and an unidentified mechanism, affected by cyclosporin A, in host defence to renal infection. PMID- 3040068 TI - The effect of PUVA treatment on acid hydrolases in human polymorphonuclear leukocytes. AB - The activity of intracellular acid hydrolases in polymorphonuclear leukocytes (PMNL) from psoriatic patients and normal control subjects was determined. No significant differences between healthy and psoriatic individuals were detected, but a slight decrease in acid hydrolase activity was found in PMNL of psoriasis patients during PUVA therapy. PUVA treatment of PMNL in vitro at intensities that may be achieved in situ in the epidermis led to intracellular inactivation of acid hydrolases, which was not due to secretion of the enzymes or cell damage. The decrease in PMNL hydrolase activity appeared to be evoked by PUVA-generated reactive oxygen species because reduced glutathione prevented this decrease. The activity of free extracellular acid hydrolases was not affected by PUVA, and the superoxide production of PUVA-treated PMNL was increased. These results suggest that intracellular inactivation of acid hydrolases and possibly other lysosomal enzymes in PMNL or monocytes infiltrating the epidermis may contribute to the antipsoriatic activity of PUVA therapy. PMID- 3040067 TI - Chlamydia trachomatis pneumonia in the immune, athymic and normal BALB mouse. AB - This paper compares the histopathology of pneumonia due to murine Chlamydia trachomatis (MoPn, mouse pneumonitis agent) in susceptible athymic nude mice (nu/nu), resistant heterozygous littermates (nu/+) and very resistant immunized nu/+ mice. While all groups had an early heterophil response, successful host defence correlated with the presence of large numbers of plasma cells, lymphocytes, monocytes, and lipid laden macrophages. Reticulate bodies were seen in all groups, predominantly in type I alveolar epithelial cells. By 24 h in the immune nu/+ group, no intact organisms were visible. Optimal control of infection was thus rapid and not clearly related to heterophils. These studies show that the histopathology of chlamydial infection may be quite atypical in the immunocompromised host, mononuclear cells seem critical in host defence, and B cell activation with plasma cell infiltration is dependent on intact T cell function in this model. PMID- 3040069 TI - Investigation of adrenomedullary function in atopic dermatitis. AB - Adrenomedullary function in patients with atopic dermatitis was assessed by measurement of plasma levels of catecholamines (adrenalin and noradrenalin) and cyclic AMP in response to the stimuli of standing after lying supine, and a 5-min infusion of histamine in the standing position (i.e. histamine plus standing). Plasma clearance of adrenalin was examined by measurement of plasma catecholamine and cyclic AMP levels following a 15-min intravenous infusion of adrenalin in the supine position. Resting plasma levels of adrenalin, cyclic AMP and noradrenalin were not statistically different in atopic patients and normal controls. Standing or intravenous infusion of histamine in the standing position caused a rise in plasma catecholamine levels. Plasma adrenalin, cyclic AMP and noradrenalin levels in response to these stimuli and the rate of clearance of exogenous adrenalin from the plasma were not significantly different in patients with atopic dermatitis and in normal subjects. PMID- 3040070 TI - Human parvovirus infection with arthropathy. PMID- 3040071 TI - Interaction of urokinase with specific receptors abolishes the time of commitment to terminal differentiation of murine erythroleukaemia (Friend) cells. AB - We have shown the presence of specific receptors for human urokinase on the surface of mouse erythroleukaemic cells. The receptor number increases when undifferentiated cells undergo hexamethylene-bisacetamide induced differentiation, while affinity between receptor and ligand does not change. A monoclonal antibody against the 17,500 mol wt fragment of the non-catalytic chain of urokinase (A chain) inhibits the specific binding, as we have previously shown in A431 cells. We have found that, upon the simultaneous addition of both urokinase or the A chain and hexamethylenebisacetamide, commitment is initiated immediately without the lag shown by control cultures of undifferentiated cells treated with the low molecular weight inducer alone. A series of mechanisms possibly involved in the action of urokinase on mouse erythroleukaemic differentiation are also discussed. PMID- 3040073 TI - Metastatic germ cell tumour associated with XY gonadal dysgenesis: successful chemotherapy. Case report. PMID- 3040072 TI - Histological types of lung cancer among smelter workers exposed to arsenic. AB - The histological distribution of lung cancer was investigated in 93 men who had worked at a Swedish smelter with high levels of arsenic. A comparison was made with a group of 136 patients with lung cancer from the county where the smelter was located. Company records provided information on occupational exposure and data on smoking habits were obtained from a next of kin of each subject. No pronounced differences in the histological types of lung carcinomas between smelter workers and the reference group could be seen for smokers. Some analyses indicated an increased proportion of adenocarcinomas among the smelter workers, which confirmed earlier data, but these findings were difficult to interpret. Cases among smelter workers who had never smoked showed a histological distribution resembling that in smokers, indicating that the work environment at the smelter and smoking had a similar influence on the risk for different types of lung cancer. PMID- 3040074 TI - A retrospective review of adenocarcinoma-in-situ and glandular atypia of the uterine cervix. AB - Between 1978 and 1985, 19 cases of adenocarcinoma-in-situ and 12 cases of glandular atypia have been identified at the Birmingham and Midland Hospital for Women. In 19 cases an associated dysplastic squamous element was identified, 20 of 28 pre-diagnosis smears correctly predicted a glandular lesion, 5 of 17 colposcopically directed biopsies predicted the findings in a larger biopsy (cone biopsy or hysterectomy). Colposcopy provided no additional information with regard to diagnosis. Twelve of 13 patients managed by cone biopsy and 16 of 17 by abdominal hysterectomy have been treated successfully as defined by subsequent normal cytology. PMID- 3040076 TI - The vitamin D status of East Indian Punjabi immigrants to Canada. AB - 1. Serum 25-hydroxyvitamin D (25-OHD), calcium and alkaline phosphatase (EC 3.1.3.1) levels and vitamin D intakes (from 3 d weighed dietary records) were determined in a cohort of fifty-nine male East Indian Punjabi immigrants (37.7 (SD 10.5) years) and fifty-four females (33.3 (SD 7.4) years). 2. Females had somewhat lower mean serum 25-OHD levels (12.3 (SD 5.0) v. 14.2 (SD 5.1) ng/ml, P less than 0.05) and serum Ca levels (88 (SD 8) v. 91 (SD 6) mg/l) than males (P less than 0.05) whereas serum alkaline phosphatase values (males 167 (SD 63), females 169 (SD 43) IU/l) and dietary vitamin D intakes (males 3.5 (SD 1.8), females 3.3 (SD 2.0) micrograms/d) were similar. 3. 22% of the females and 12% of the males had serum 25-OHD levels below 9.0 ng/ml but none had serum 25-OHD levels within the range associated with clinically overt disease. 4. In the males, serum 25-OHD levels were negatively correlated with dietary fibre intakes (g/d; r -0.29; P less than 0.05). 5. Multiple-regression analysis indicated that log serum 25-OHD levels were not related to dietary vitamin D intakes. Instead they were associated with sex and dietary fibre intakes (g/MJ) (F 3.71; P = 0.03). These two variables explained 8% of the variance. PMID- 3040075 TI - Comparison of fibre digestion and digesta retention time between rabbits, guinea pigs, rats and hamsters. AB - 1. Digestive efficiencies of fibre components and retention time of digesta in the whole gut and in the large intestine were measured in rabbits, guinea-pigs, hamsters and rats when given a lucerne (Medicago sativa)-containing diet. 2. Co EDTA and chromium-mordanted cell-wall constituents of Italian ryegrass (Lolium multiflorum L.) were used as liquid- and solid-phase markers respectively. Both markers were mixed with the experimental diet and given after digestion trials. 3. Mean retention times of each marker were calculated from time-course changes in concentrations of the markers in faeces. The mean retention times of the markers in the large intestine were calculated from exponential slopes fitted to the time-course changes of faecal concentrations of the markers. 4. The digestibilities of crude fibre, neutral-detergent fibre and acid-detergent fibre were highest in the guinea-pigs, followed by the hamsters, and lowest in the rabbits and rats. 5. The mean retention times of Cr in the whole tract were longer in the larger animals and shortest in the hamsters. The mean retention times of Cr in the large intestine were longest in the guinea-pig followed by the hamsters and the rats. The rabbits had an extremely short retention time of Cr in the large intestine. 6. These results suggest that the retention time of solid digesta in the large intestine can explain the difference in the digestive efficiencies of fibre components amongst non-ruminant small herbivores whereas retention of digesta in the whole gut is not related to the digestibility of fibre components. PMID- 3040077 TI - The effect of vegetables and beet fibre on the absorption of zinc in humans from composite meals. AB - 1. The absorption of zinc in humans from composite meals, was determined by extrinsic labelling of the meals with 65Zn and measurement of the whole-body retention of the radioisotope. 2. Low-Zn (mean 25 mumol) chicken meals with 150 g white bread or 225 g potatoes, carrots, turnips, cabbage or green peas were studied. The effect of a beet-pulp-fibre preparation used as a breakfast cereal, in bread and as a meat extender on Zn absorption was also studied. 3. The mean percentage absorption from the chicken meals with white bread, carrots and cabbage was significantly different from the meals with potatoes, turnips and green peas. When the amount of Zn in the meals was taken into account a slightly higher absorption was observed from the white-bread meal compared with the meals with potatoes and cabbage, while no differences were seen between the vegetable meals. 4. The beet-pulp-fibre preparation did not affect the extent of Zn absorption when used as a meat extender. The absorption of Zn was higher when the beet fibre was included in bread than when used as muesli. 5. The results obtained suggest that, besides the low-Zn content in vegetables, a large intake of vegetables or a pure-vegetable-fibre preparation has no significant effect on Zn availability from animal-protein-based meals. PMID- 3040078 TI - Cardiovascular beta-adrenoceptor sensitivity of undernourished subjects. AB - 1. Eleven normal-weight male subjects (weight/height2 (W/H2) 19.0-22.5) from a good socio-economic background and on ad lib. food intake, and eight undernourished male labourers (W/H2 16.4-18.6) on low energy intakes, were studied. 2. Comparison of cardiovascular responses to increasing single doses of the beta-agonist, isoproterenol, showed a significantly greater positive chronotropic dose response in the undernourished subjects. 3. Cardiovascular responses to head-up tilt were similar in both the normal-weight and undernourished subjects. 4. Undernourished individuals may show an increase in beta-adrenoceptor sensitivity which may be akin to a denervation type of supersensitivity as a result of a nutrition-related reduction in sympathetic activity. PMID- 3040079 TI - The effect of dietary protein source and guar gum on gastrointestinal growth and enteroglucagon secretion in the rat. AB - 1. Male Wistar rats (approximately 100 g) were given fibre-free semi-synthetic diets containing either casein or albumin (168 g/kg diet) as the protein source with or without guar gum (75 g/kg diet) (casein, albumin, casein guar gum and albumin-guar gum groups). 2. Small intestinal length, weights of caecal tissue and contents and plasma enteroglucagon concentration were significantly increased in guar-gum-fed animals compared with the fibre-free groups. 3. Total caecal weight and plasma enteroglucagon concentration were higher in the albumin-guar gum group compared with the casein-guar gum group. The weights of caecal tissue and contents were significantly increased in rats given the fibre-free albumin diet compared with those consuming a similar diet with casein as the protein source, although daily food intake tended to be lower. 4. It is concluded that the effect of materials classed as dietary fibre may be significantly influenced by the non-polysaccharide component of the diet, and that such interactions may influence both the growth and endocrine activity of the gastrointestinal tract. PMID- 3040080 TI - Extended X-ray absorption fine structure of copper in CuA-depleted, p (hydroxymercuri)benzoate-modified, and native cytochrome c oxidase. AB - Cytochrome c oxidase contains four redox-active metal centers: two heme irons, cytochromes a and a3, and two copper ions, CuA and CuB. Due to the paucity of spectroscopic signatures for both copper sites in cytochrome c oxidase, the ligands and structures for these sites have remained ambiguous. The specific depletion of CuA from the p-(hydroxymercuri)benzoate- (pHMB-) modified cytochrome c oxidase recently reported [Gelles, J., & Chan, S. I. (1985) Biochemistry 24, 3963-3972] is herein described. Characterization of this enzyme shows that the structures of the remaining metal centers are essentially unperturbed by the CuA modification and depletion (P. M. Li, J. Gelles, and S. I. Chan, unpublished results). Copper extended X-ray absorption fine structure (EXAFS) measurements on the CuA-depleted cytochrome c oxidase reveal coordination of three (N, O) ligands and one (S, Cl) ligand at the CuB site. Comparison of EXAFS results obtained for the CuA-depleted, pHMB-modified, and "unmodified control" enzymes has allowed the deconvolution of the EXAFS in terms of the inner coordination spheres for CuA as well as CuB. On the basis of these data, it is found that the structure for the CuA site is consistent with two (N, O) ligands and two S ligands. PMID- 3040081 TI - Distance estimate of the active center of D-beta-hydroxybutyrate dehydrogenase from the membrane surface. AB - D-beta-Hydroxybutyrate dehydrogenase (EC 1.1.1.30) is a membrane-bound, lipid requiring enzyme which has a reactive sulfhydryl in the vicinity of the active center. The spin-probe-spin-label technique has been used to estimate the distance of separation of the reactive sulfhydryl of D-beta-hydroxybutyrate dehydrogenase from the bilayer surface. The reactive sulfhydryl of the enzyme was derivatized with the maleimide spin-label reagent 4-maleimido-2,2,6,6 tetramethylpiperidinyl-1-oxy in the presence of the cofactor NAD+. The derivatized enzyme, inserted (inlaid orientation) into phospholipid vesicles, was titrated with spin probes, either Mn2+ or Gd3+, until the spin-label EPR spectrum was reduced in amplitude to its residual (limiting) value. From this limiting amplitude, the dipolar interaction coefficient was obtained, which is related to the reciprocal of the distance to the sixth power. The radial distances of closest approach of the paramagnetic Mn2+ and Gd3+ ions to the spin-label nitroxide on the enzyme were found to be 18 and 16 A, respectively. These calculated distances were in accord with those determined by comparison with a phosphatidylcholine calibration system having 2,2-dimethyloxazolidinyl-1-oxy spin labels located at selected positions along the sn-2 fatty acyl chain. Since the distal nitroxide moiety of the maleimide spin-label (17 A from the bilayer surface) is 8 A from the sulfhydryl addition site, the two limiting distances of immersion of the reactive sulfhydryl within the bilayer are 9 and 25 A. The shorter distance is considered more compatible with facile access of the coenzyme to the active site of the enzyme. PMID- 3040082 TI - Interaction of polymyxin B nonapeptide with anionic phospholipids. AB - The interaction of polymyxin B nonapeptide (PMBN) and polymyxin B (PMB) with the anionic phospholipids phosphatidylserine (PS), dipalmitoylphosphatidylglycerol (DPPG), dipalmitoylphosphatidic acid (DPPA), and 1:1 mixtures (w/w) of DPPA and distearoylphosphatidylcholine (DSPC) was studied by calorimetry, electron spin resonance, and fluorescence spectrometry, electron microscopy, and fusion and leakage assays. The phase transition temperatures of DPPA and DPPG were very similar when bound to PMB or PMBN, indicating that the lipids are in a similar state when bound to the cationic peptides. Both PMB and PMBN caused the interdigitation of DPPG bilayers, suggesting that the penetration of hydrophobic side chains from a peptide bound electrostatically on the surface is sufficient to induce this phenomenon. Stopped-flow experiments revealed that PMBN and PMB induced the fusion of small unilamellar PS and large unilamellar DPPA-DSPC vesicles. The aggregation of vesicles was found to be diffusion-controlled process; the subsequent fusion took place with a frequency of 10(2)-(5 X 10(2] s 1 for small vesicles and 1-100 s-1 for large vesicles. The freeze-fracture replicas of the PMB-treated vesicles displayed 12-50-nm depressions on several superimposed bilayers, indicating the formation of stable lipid-PMB domains. Since the incubation with PMBN produced similar depressions only if the specimens were fixed, PMBN-induced domain formation seems to be a reversible rapid process. The differences in the phospholipid-peptide interactions are correlated with the differences in the physiological action of the antibiotic PMB and the nonbactericidal PMBN on the cell envelope of Gram-negative bacteria. PMID- 3040083 TI - Directly observed 15N NMR spectra of uniformly enriched proteins. AB - The proteins cytochrome c2, cytochrome c', and ribulosebisphosphate carboxylase/oxygenase from Rhodospirillum rubrum were enriched in 15N by growth of the organism on 15NH4Cl. The proteins were purified to homogeneity and studied by 15N NMR. Longitudinal and transverse relaxation times as well as the nuclear Overhauser effects were determined for various groups of the proteins which vary in molecular weight from 13,000 to 114,000. The values of these parameters for the amide resonances or for groups thought to be rigid were consistent with the molecular weights of the proteins. Relaxation times of the amino-terminal alpha amino groups and the side chain nitrogen atoms of arginine and lysine were consistent with much more rapid motion. Nitrogen atoms having bound protons were generally found to be decoupled from the protons by chemical exchange. Demonstrable 1H-15N coupling was taken as an indication that exchange was hindered, either by hydrogen bonding interactions or by inaccessibility of the group to solvent. Histidine side chain nitrogen atoms, which experience a large chemical shift upon protonation/deprotonation, were often found to be broadened beyond detectability by chemical exchange and tautomerization. Strategies for improving sensitivity and for obtaining specific peak assignments are also discussed. PMID- 3040084 TI - Palindromic octa- and dodecanucleotides containing 2'-deoxytubercidin: synthesis, hairpin formation, and recognition by the endodeoxyribonuclease EcoRI. AB - Octa- and dodecanucleotides containing 2'-deoxytubericidin within the endodeoxyribonuclease EcoRI recognition fragment d(GAATTC) have been prepared by solid-phase synthesis. Whereas octamers as well as dodecamers with a "random" flanking region formed duplexes in aqueous solution, the dodecamer d(CGCGAATTCGCG) and isosterically modified oligomers thereof showed a strong tendency of hairpin formation. Due to this, cleavage with the endodeoxyribonuclease EcoRI was strongly decreased. In contrast, d(GTAGAATTCTAC) was easily cleaved by the enzyme. Single replacement of one of the dA residues by 2'-deoxytubercidin within the recognition sequence decreased the cleavage velocity but retained specificity. Twofold modification prevents cleavage of the oligomer. This implies that both N-7 purine nitrogens are proton acceptor sites for the endodeoxyribonuclease EcoRI. PMID- 3040085 TI - Structure of the oxalate-ATP complex with pyruvate kinase: ATP as a bridging ligand for the two divalent cations. AB - The 2 equiv of divalent cation that are required cofactors for pyruvate kinase reside in sites of different affinities for different species of cation [Baek, Y. H., & Nowak, T. (1982) Arch. Biochem. Biophys. 217, 491-497]. The intrinsic selectivity of the protein-based site for Mn(II) and of the nucleotide-based site for Mg(II) has been exploited in electron paramagnetic resonance (EPR) investigations of ligands for Mn(II) at the protein-based site. Oxalate, a structural analogue of the enolate of pyruvate, has been used as a surrogate for the reactive form of pyruvate in complexes with enzyme, Mn(II), Mg(II), and ATP. Addition of Mg(II) to solutions of enzyme, Mn(II), ATP, and oxalate sharpens the EPR signals for the enzyme-bound Mn(II). Superhyperfine coupling between the unpaired electron spin of Mn(II) and the nuclear spin of 17O, specifically incorporated into oxalate, shows that oxalate is bound at the active site as a bidentate chelate with Mn(II). Coordination of the gamma-phosphate of ATP to this same Mn(II) center is revealed by observation of superhyperfine coupling form 17O regiospecifically incorporated into the gamma-phosphate group of ATP. By contrast, 17O in the alpha-phosphate or in the beta-phosphate groups of ATP does not influence the spectrum. Experiments in 17O-enriched water show that there is also a single water ligand bound to the Mn(II). These data indicate that ATP bridges Mn(II) and Mg(II) at the active site. A close spacing of the two divalent cations is also evident from the occurrence of magnetic interactions for complexes in which 2 equiv of Mn(II) are present at the active site.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3040086 TI - Lignin peroxidase: resonance Raman spectral evidence for compound II and for a temperature-dependent coordination-state equilibrium in the ferric enzyme. AB - Resonance Raman (RR) spectroscopy of lignin peroxidase (ligninase, dairylpropane oxygenase) from the basidiomycete Phanerochaete chrysosporium suggests two different coordination states for the native ferric enzyme. Evidence for a high spin, hexacoordinate ferric protoporphyrin IX was presented by Andersson et al. [Andersson, L. A., Renganathan, V., Chiu, A.A., Loehr, T. M., & Gold, M. H. (1985) J. Biol. Chem. 260, 6080-6087], whereas Kuila et al. [Kuila, D., Tien, M., Fee, J. A., & Ondrias, M. R. (1985) Biochemistry 24, 3394-3397] proposed a high spin, pentacoordinate ferric system. Because the two RR spectral studies were performed at different temperatures, we explored the possibility that lignin peroxidase might exhibit temperature-dependent coordination-state equilibria. Resonance Raman results presented herein indicate that this hypothesis is indeed correct. At or near 25 degrees C, the ferric iron of lignin peroxidase is predominantly high spin, pentacoordinate; however, at less than or equal to 2 degrees C, the high-spin, hexacoordinate state dominates, as indicated by the frequencies of well-documented spin- and coordination-state marker bands for iron protoporphyrin IX. The temperature-dependent behavior of lignin peroxidase is thus similar to that of cytochrome c peroxidase (CCP). Furthermore, lignin peroxidase, like horseradish peroxidase (HRP) and CCP, clearly has a vacant coordination site trans to the native fifth ligand at ambient temperature. High frequency RR spectra of compound II of lignin peroxidase are also presented. The observed shifts to higher frequency for both the oxidation-state marker band v4 and the spin- and coordination-state marker band v10 are similar to those reported for the compound II forms of HRP and lactoperoxidase and for ferryl myoglobin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3040087 TI - Inhibition of 3(17)beta-hydroxysteroid dehydrogenase from Pseudomonas testosteroni by steroidal A ring fused pyrazoles. AB - Several 2,3- and 3,4-steroidal fused pyrazoles have been investigated as potential inhibitors of NAD(P)H-dependent steroid oxidoreductases. These compounds are proven to be potent, specific inhibitors for 3(17) beta hydroxysteroid dehydrogenase from Pseudomonas testosteroni with Ki values of 6 100 nM. In contrast, the activities of 3 alpha,20 beta-hydroxysteroid dehydrogenase from Streptomyces hydrogenans, steroid 5 alpha-reductase from rat prostate, and 3 alpha-hydroxysteroid dehydrogenase from rat liver were unaffected by micromolar concentrations of these compounds. Product and dead-end inhibition studies indicate an ordered association to the beta-dehydrogenase with the cofactor binding prior to substrate or inhibitor. From the results of double inhibition experiments, it is proposed that inhibition occurs through formation of an enzyme-NAD+-inhibitor ternate. On the basis of pH profiles of Vm/Km, Vm, and 1/Ki and of absorbance difference spectra, a hypothetical mechanism of inhibition by the steroidal pyrazoles, drawn by analogy from the inhibition of liver alcohol dehydrogenase by alkylpyrazoles [Theorell, H., & Yonetani, T. (1963) Biochem. Z. 338, 537-553; Andersson, P., Kvassman, J. K., Lindstrom, A., Olden, B., & Pettersson, G. (1981) Eur. J. Biochem. 113, 549-554], is reconsidered. The pH studies and enzyme modification experiments by diethyl pyrocarbonate suggest the involvement of histidine in binding of the inhibitor. A modified proposal for the structure of the enzyme-NAD+-steroidal pyrazole complex is proposed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3040088 TI - Low-temperature electron paramagnetic resonance study of the ferricytochrome c cardiolipin complex. AB - The electron paramagnetic resonance spectrum of the ferricytochrome c complex with cardiolipin was observed at temperatures below 20 K. For the low-spin iron(III) heme system complexed with the negatively charged lipid, the tetragonal and rhombic ligand field parameters (delta/lambda = 3.58, V/lambda = 1.82) differ significantly from those (delta/lambda = 2.53, V/lambda = 1.49) of the free ferricytochrome c sample. The g values of the complex (gx = 1.54 +/- 0.02, gy = 2.26 +/- 0.01, gz = 3.02 +/- 0.01) are compared to the values for free ferricytochrome c (gx = 1.25 +/- 0.02, gy = 2.25 +/- 0.01, gz = 3.04 +/- 0.01). Spectral alterations are interpreted in terms of the ligand field changes induced within the heme group by association with the negatively charged phosphoglyceride. PMID- 3040089 TI - Separate domains of the insulin receptor contain sites of autophosphorylation and tyrosine kinase activity. AB - We have studied the structure and function of the solubilized insulin receptor before and after partial proteolytic digestion to define domains in the beta subunit that undergo autophosphorylation and contain the tyrosine kinase activity. Wheat germ agglutinin purified insulin receptor from Fao cells was digested briefly at 22 degrees C with low concentrations (5-10 micrograms/mL, pH 7.4) of trypsin, staphylococcal V8 protease, or elastase. Autophosphorylation of the beta-subunit was carried out before and after digestion, and the [32P]phosphoproteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, detected by autoradiography, and analyzed by tryptic peptide mapping by use of reverse-phase high-performance liquid chromatography. Mild trypsin digestion reduced the apparent molecular mass of the beta-subunit from 95 to 85 kDa, and then to 70 kDa. The 85-kDa fragment was not immunoprecipitated by an antibody directed against the C-terminal domain of the beta-subunit (alpha Pep 1), indicating that this region of the receptor was lost. The 85-kDa fragment contained about half of the [32P]phosphate originally found in the beta-subunit, and tryptic peptide mapping showed that two major tryptic phosphopeptides (previously called pY2 and pY3) were removed. Three other tryptic phosphopeptides (pY1, pY1a, and pY4) were found in the 85- and 70-kDa fragments. Treatment of the intact receptor with staphylococcal V8 protease also converted the beta-subunit to an 85-kDa fragment that did not bind to alpha Pep-1, contained about 50% of the initial radioactivity, and lacked pY2 and pY3. Elastase rapidly degraded the receptor to inactive fragments between 37 and 50 kDa.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3040090 TI - The proton-pumping site of cytochrome c oxidase: a model of its structure and mechanism. AB - Cytochrome c oxidase is an electron-transfer driven proton pump. In this paper, we propose a complete chemical mechanism for the enzyme's proton-pumping site. The mechanism achieves pumping with chemical reaction steps localized at a redox center within the enzyme; no indirect coupling through protein conformational changes is required. The proposed mechanism is based on a novel redox-linked transition metal ligand substitution reaction. The use of this reaction leads in a straightforward manner to explicit mechanisms for achieving all of the processes previously determined (Blair, D.F., Gelles, J. and Chan, S.I. (1986) Biophys. J. 50, 713-733) to be needed to accomplish redox-linked proton pumping. These processes include: (1) modulation of the energetics of protonation/deprotonation reactions and modulation of the energetics of redox reactions by the structural state of the pumping site; (2) control of the rates of the pump's redox reactions with its electron-transfer partners during the turnover cycle (gating of electrons); and (3) regulation of the rates of the protonation/deprotonation reactions between the pumping site and the aqueous phases on the two sides of the membrane during the reaction cycle (gating of protons). The model is the first proposed for the cytochrome oxidase proton pump which is mechanistically complete and sufficiently specific that a realistic assessment can be made of how well the model pump would function as a redox linked free-energy transducer. This assessment is accomplished via analyses of the thermodynamic properties and steady-state kinetics expected of the model. These analyses demonstrate that the model would function as an efficient pump and that its behavior would be very similar to that observed of cytochrome oxidase both in the mitochondrion and in purified preparations. The analysis presented here leads to the following important general conclusions regarding the mechanistic features of the oxidase proton pump. (1) A workable proton-pump mechanism does not require large protein conformational changes. (2) A redox linked proton pump need not display a pH-dependent midpoint potential, as has frequently been assumed. (3) Mechanisms for redox-linked proton pumps that involve transition metal ligand exchange reactions are quite attractive because such reactions readily lend themselves to the linked gating processes necessary for proton pumping. PMID- 3040091 TI - The effect of detergents on bovine cytochrome c oxidase: a kinetic approach. AB - (1) Investigation of the relationship between the detergent concentration and steady-state and pre-steady-state kinetics of cytochrome c oxidase proved to be a valid approach in the study of protein-detergent interaction. (2) Laurylmaltoside, sodium cholate and Triton X-100 influenced the kinetics of cytochrome c oxidase cooperatively at detergent concentrations near their critical micelle concentration. This mode of interaction reflects disaggregation of the oxidase as a result of cooperative binding of the detergent. (3) Addition of increasing concentrations of Tween-80 to the aggregated enzyme caused a more gradual decrease in aggregation of the oxidase, which did not result in a change in activity of the enzyme. This suggests that aggregation of cytochrome c oxidase occurs in a highly regular manner in which no catalytic sites are shielded off. (4) Oxidase aggregates present at detergent concentrations below the critical micelle concentration of laurylmaltoside and Triton X-100 showed considerable activity. Their kinetics were equal to those of the oxidase in Tween-80, suggesting that the protein molecules are aligned in a similar way in all oligomers. Aggregates present in low concentrations of sodium cholate showed turnover rates that were twice as low as those observed with other aggregates. (5) Solubilisation of the oxidase by sodium cholate or Triton X-100 resulted in almost complete inhibition of enzymic activity, whereas the association rate of ferrocytochrome c was almost equal to that found for monomeric oxidase in laurylmaltoside. These results are in agreement with a mixed-type inhibition. PMID- 3040092 TI - Bovine cytochrome c oxidases, purified from heart, skeletal muscle, liver and kidney, differ in the small subunits but show the same reaction kinetics with cytochrome c. AB - (1) Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate of purified cytochrome c oxidase preparations revealed that bovine kidney, skeletal muscle and heart contain different cytochrome c oxidase isoenzymes, which show differences in mobility of the subunits encoded by the nuclear genome. No differences in subunit pattern were observed between the oxidase preparations isolated from kidney and liver. (2) The kinetics of the steady-state reactions between bovine ferrocytochrome c and the four types of bovine cytochrome c oxidase preparation were compared under conditions of both high- and low-ionic strength. Also the pre-steady-state kinetics were studied. Only minor differences were observed in the electron-transfer activity of the isoenzymes. Thus, our experiments do not support the notion that the subunits encoded by the nuclear genome act as modulators conferring different activities to the isoenzymes of cytochrome c oxidase. (3) The cytochrome c oxidase preparation from bovine skeletal muscle was found to consist mainly of dimers, whereas the enzymes isolated from bovine kidney, liver and heart were monomeric. PMID- 3040093 TI - Electron flow and heme-heme interaction between cytochromes b-558, b-595 and d in a terminal oxidase of Escherichia coli. AB - The ESR signals of the cytochromes in the Escherichia coli terminal oxidase cytochrome d complex were studied at cryogenic temperature. The intensities and g values of the rhombic high-spin signals changed when the electronic state of cytochrome d was changed from the oxidized state to the reduced or oxygen-binding or CO-binding state. These rhombic signals were therefore assigned to cytochrome b-595, which is located near cytochrome d in the oxidase complex. This assignment was supported by the finding that the Em value of the rhombic signals differed from that of cytochrome d (Hata, A. et al. (1985) Biochim. Biophys. Acta 810, 62 72). Photolysis and ligand-exchange experiments with the reduced CO complex of the oxidase were performed in the presence of oxygen at -140 degrees C. The ESR spectra of three intermediate forms trapped by controlled low temperatures were detected. These forms were designated as the oxygen-binding intermediate I (ESR silent), oxygen-binding intermediate II (giving ESR signals at g = 6.3, 5.5 and 2.15), and oxygen-binding intermediate III (giving signals at g = 6.3, 5.5 and 6.0). From these results, electron flow in the cytochrome d complex is proposed to proceed in the order, cytochrome b-558----cytochrome b-595----cytochrome d--- O2. A model of the mechanism of four-electron chemistry for oxidation of ubiquinol-8 and formation of H2O by the cytochrome d complex is presented. PMID- 3040094 TI - A long chain spin label for glycosphingolipid studies: transbilayer fatty acid interdigitation of lactosyl ceramide. AB - 16-Carbon and 18-carbon fatty acids with covalently attached nitroxide free radicals have seen wide usage in membrane studies of phospholipid dynamics, orientation, and associations. However, they are inadequate for dealing with some very important questions that relate to glycosphingolipids. We report here the synthesis of a long chain (24-carbon) spin-labelled fatty acid designed for such problems. We have used both the new 24-carbon and the more conventional 18-carbon spin-labelled fatty acids to replace the natural fatty acid of lactosyl ceramide so that we may begin to compare short and long chain derivatives to analyse the molecular basis of their functional differences. Spectra seen are consistent with the view that in a bilayer host matrix the methyl end of the long fatty acid crosses the hydrophobic membrane center and interdigitates with fatty acids of phospholipids of the opposing monolayer. PMID- 3040095 TI - Effect of oxygen free radicals on ubiquinone in aqueous solution and phospholipid vesicles. AB - The purpose of this study was to evaluate the direct effect of oxygen free radicals produced by ultrasonic irradiation on ubiquinone and to compare the efficiency with which the antioxidant can compete with these radicals when it is both in aqueous solution and within the lipid bilayer. The main product obtained after insonation of aqueous solutions of ubiquinone-0 was ubiquinol, moreover some degradation occurred. The direct electron donor responsible for most of the ubiquinol generated by ultrasonic irradiation appeared to be superoxide radical. Addition reactions of hydroxyl radicals with aromatic ring structure led probably to degradation products of ubiquinone, which were not identified. Experiments were also performed to evaluate the efficiency with which ubiquinone-3 could react with oxygen radicals when it was within the lipid bilayer. The effect of presence or absence of a net surface charge was studied selecting a suitable bilayer including dimyristylphosphatidic acid or stearylamine in uncharged dimyristylphosphatidylcholine vesicles. In these systems hydroxyl radicals did not represent a potential danger for the antioxidant, the reaction between superoxide and ubiquinone-3 instead was significant only in positively charged membranes and gave rise to ubiquinol. It is suggested that ubiquinone acts as an antioxidant by stopping the propagation reaction. PMID- 3040096 TI - Interactions of mitochondrial precursor protein apocytochrome c with phosphatidylserine in model membranes. A monolayer study. AB - (1) The interaction of apocytochrome c with different molecular species of phosphatidylserine was studied using monolayers at constant surface area or constant surface pressure. The protein inserted readily into dioleoylphosphatidylserine monolayers up to a limiting pressure of 50 mN/m, whereas the interaction decreased with increasing molecular packing of the phosphatidylserine species, indicating the importance of the hydrophobic core of the lipid layer for the interaction. (2) The high affinity of apocytochrome c for dioleoylphosphatidylserine is indicated by the low Kd of 0.017 microM. There is little or no interaction with phosphatidylcholines. The importance of charge interactions is underlined by its ionic strength and pH dependency. (3) Experiments using 14C-labelled apocytochrome c indicate that cholesterol can enhance the protein binding. (4) It was demonstrated that apocytochrome c monomers penetrate the monolayer whereas oligomers can be formed in an adsorbed layer and washed off without changing the surface pressure. Preincubation of apocytochrome c in 3 M guanidine, to obtain the monomeric form, was essential to measure the full effect of interfacial interaction. (5) The molecular area of apocytochrome c changed from 1200-1300 A2/molecule in the absence of lipid to 700 900 A2/molecule after penetration of dioleoylphosphatidylserine monolayers. (6) Apocytochrome c-dioleoylphosphatidylserine interactions are only possible when the monolayer is approached from the subphase. It is concluded that the charge interactions are required for binding and penetration of the protein. PMID- 3040097 TI - Prostaglandin E2 directly protects isolated rat gastric surface cell membranes against bile salts. AB - Rat gastric surface cell membranes were prepared and the effect of taurocholic acid assessed by ESR spectroscopy using the 16-doxylstearic acid spin label. Taurocholic acid increased the polar part of the spectra, indicating an augmented amount of spin label molecules with a polar environment. Concomitantly, mobility of the spin label molecule was augmented. The effect of taurocholic acid was completely prevented by the previous addition of prostaglandin E2. This suggests a direct protective efficiency of prostaglandin E2 on rat gastric surface cell membranes without the metabolic participation of intact cells. PMID- 3040098 TI - The influence of dolichols on fluidity of mouse synaptic plasma membranes. AB - Dolichols are isoprenologues which constitute an important component of biological membranes. However, an understanding of the effects of dolichols on the organization and dynamics of biological membranes has not been forthcoming. The experiments reported here are aimed at understanding the effects of dolichols on the physical properties of mouse brain synaptic plasma membranes. The effect of dolichols incorporated into mouse brain synaptic plasma membranes on fluorescent and electron spin resonance probes sensing the hydrophobic core differed from that of probes reporting closer to the surface of membrane bilayers. Dolichols significantly (P less than 0.01) lowered the polarization, limiting anisotropy, and order parameter of diphenylhexatriene in synaptic plasma membranes and liposomes extracted from synaptic plasma membranes, without changing the rotational relaxation time. Similarly, dolichol increased the fluidity reported by 16-doxylstearic acid in synaptic plasma membranes or liposomes extracted from synaptic plasma membranes. In contrast, dolichols exerted no effect on those properties for trans-parinaric acid or 5-doxylstearic acid in synaptic plasma membranes or liposomes derived therefrom. Dolichols can dramatically alter the structure and dynamics of lipid motion in synaptic plasma membranes and these effects are dependent on the location of the probe in the membrane. PMID- 3040099 TI - Influence of DNA length on spermine-induced condensation. Importance of the bending and stiffening of DNA. AB - Using DNA restriction fragments of 258 to 4362 base-pairs, we have investigated the influence of the DNA length on the condensation process induced by spermine, with the aid of electric dichroism measurements. The 258- and 436 bp fragments condensed into rod-like particles, while the fragments of 748 bp or more condensed into torus-shaped particles. Our results suggest that a DNA molecule longer than the circumference of the toroids observed previously (680 bp) is required to serve as a nucleus for the growth of the condensed particles. The toroids were more stable in the electric field than the rod-shaped particles, suggesting that rapid fluctuations of the bound spermine counterions can provide one of the main attractive forces yielding to the condensation process. Relaxation time data for the 436 bp fragment revealed that the structure of DNA was altered at a spermine concentration as low as one-tenth of that required for condensation: the DNA became bent in the presence of spermine. Moreover, the field strength dependence of the relaxation times, as well as the fitting of the decay curves at 12.5 kV/cm, showed an increase of the stiffness of the DNA double helix upon spermine addition. We estimated that, in the case of DNA condensation by spermine, a decrease in the measured persistence length may occur, irrespective of the DNA flexibility, owing to the bending of the DNA molecule. PMID- 3040100 TI - The ubiquitinated histone species are enriched in histone H1-depleted chromatin regions. AB - Bovine thymus and trout testis chromatin were fractionated into regions which differed in their micrococcal nuclease accessibility and solubility properties, and the distribution of the ubiquitinated histone species among these chromatin regions was elucidated. Ubiquitinated (u) species of histones H2A and H2B were enriched in the nuclease-sensitive, low-ionic-strength, soluble fraction of both chromatins. These results indicate that the presence of ubiquitinated histones may alter nucleosome-nucleosome interactions and destabilize higher-order chromatin structures. Bovine thymus chromatin was separated into aggregation resistant, salt-soluble and aggregation-prone, salt-insoluble chromatin fractions. The aggregation-resistant chromatin fraction depleted in H1 histones was enriched in uH2A and uH2B, with uH2B showing the greater enrichment. The chromatin fragments were also stripped and reconstituted with the H1 histones prior to fractionation. The results were the same as above: uH2A and uH2B were preferentially localized in the aggregation-resistant. H1-depleted chromatin fraction, suggesting that chromatin regions enriched in ubiquitinated histone species have a reduced affinity for the H1 histones. Thus, ubiquitinated histone species may be one of the contributing factors in the differential assembly of various parts of the genome. PMID- 3040101 TI - Phosphorylation of high-mobility-group protein 14 by two specific kinases modifies its interaction with histone oligomers in free solution. AB - Chromosomal protein HMG14 can be specifically phosphorylated by the cyclic AMP dependent protein kinase at the N-terminus and by casein kinase 2 at the acidic C terminus. Under the same conditions used for HMG14, HMG17 is not significantly phosphorylated by either of the two kinases. Further, we have studied the effect of phosphorylation by these kinases on the interaction of HMG14 with histone oligomers, using chemical cross-linking. Our results indicate that the phosphorylation of HMG14 by casein kinase 2 enhances its interaction with histone oligomers in free solution, whereas a minor effect was observed by phosphorylation with cyclic AMP-dependent protein kinase. In contrast, HMG17 does not interact at all with any histone oligomer in free solution under the conditions used. To gain insight into the possible effect that phosphorylation may play in vivo, the pattern of distribution among different chromatin fractions was analysed. It was found that, although phosphorylation of HMG14 by both kinases allowed reconstitution of HMG14 to chromatin, the patterns obtained showed some slight differences. PMID- 3040102 TI - Interaction of AluI, Cfr6I and PvuII restriction-modification enzymes with substrates containing either N4-methylcytosine or 5-methylcytosine. AB - The cleavage specificity of R.Cfr6I, an isoschizomer of PvuII restriction endonuclease was determined to be 5'CAG decreases CTG and the methylation specificity of Cfr6I and PvuII methylases, 5'CAG4mCTG. Thus, M.Cfr6I and M.PvuII are new additions to the list of methylases with N4-methylcytosine specificity. Neither of the above RM enzymes acts on the substrates containing either N4 methylcytosine or 5-methylcytosine in a cognate methylation position. PMID- 3040103 TI - Molecular studies on thiaminase I. AB - The thiaminase I gene of Bacillus thiaminolyticus was cloned on a 1.6 kb DNA fragment (enzyme molecular weight 42,000), and was expressed in both Escherichia coli and Bacillus subtilis. When a selection drug was absent, the plasmid was maintained stably for approx. 100 generations in wild-type E. coli. Instability of the thiaminase gene was demonstrated in the thiamin pyrophosphate-requiring mutant of E. coli from which the plasmid was deleted rapidly. Wild-type E. coli accumulated the enzyme in its periplasm. A method for the detection of thiaminase I enzyme in SDS-polyacrylamide gel was developed. Thiaminase I of B. thiaminolyticus was found to exist in two sizes, 44 and 42 kDa, among different strains. Moreover, thiaminase of 42 kDa became approximately 41 kDa after a long term culture, most likely because of the action of proteinases. Thiaminase expressed in E. coli from a thiaminase-positive recombinant plasmid was 42 kDa, and showed the same mobility on SDS-polyacrylamide gele electrophoresis as the enzyme isolated from the young culture of the parent strain of B. thiaminolyticus used for cloning. This value was, therefore, considered to represent intact thiaminase that had escaped from the attack of bacilli proteinases. PMID- 3040104 TI - ADPribosylation of nuclear proteins labeled with [3H]adenosine: changes during the HeLa cycle. AB - Cell cycle variations in the modification of histones and nonhistones by ADPribosylation were investigated. Proteins of HeLa interphase nuclei and metaphase chromosomes were radioactively labeled in vivo with [3H]adenosine. Histones of metaphase chromosomes were extensively modified by ADPribosylation, with H2B, H2A and H4 being predominant acceptors of [3H]adenosine label. For histones of interphase nuclei from synchronized cells, the highest level of 3H labeling was observed by two-dimensional gel electrophoresis to occur in S phase. The minimum level was noted in G1 phase. ADPribosylation of histones is, however, significant during all phases of the cell cycle. These conclusions were confirmed by experiments using [32P]NAD. The results with the specific inhibitor of ADPribosylation, 3-aminobenzamide, and with snake venom phosphodiesterase indicated that the radioactive isotopes were incorporated as ADPribose. Two dimensional gels of HeLa nonhistones labeled with [3H]adenosine showed strikingly different patterns for interphase and metaphase samples. Over 100 ADPribosylated species were found for interphase nuclei, but poly(ADPribose) polymerase was the only major acceptor for metaphase chromosomes. A simple pattern was also revealed for nuclear scaffolds, with the 'lamins' and poly(ADPribose) polymerase being identifiable as modified species. PMID- 3040105 TI - The pattern of methylation in rearranged and germ-line human immunoglobulin constant mu genes. AB - Human C mu genes show two different patterns of demethylation. The first is associated with gene expression; this pattern is restricted to productive C mu alleles and is detected in leukemic and normal B-lymphocytes but not in T lymphocytes, granulocytes or fibroblasts. The second pattern, not related with gene expression, is observed in B-lymphocytes, fibroblasts and part of T lymphocytes but not in granulocytes. PMID- 3040106 TI - Structural characterization of the isoenzymatic forms of human myeloperoxidase. AB - Myeloperoxidase from human neutrophils was isolated by ion-exchange and gel filtration chromatography and shown by SDS-polyacrylamide gel electrophoresis to be comprised of alpha and beta subunits with apparent Mr values of 58,000 and 15,000, respectively. The apparent Mr of the native protein was 130,000-140,000, indicating that the holoenzyme has the quaternary structure alpha 2 beta 2. Automated Edman degradation of the separated alpha and beta subunits showed that the amino-terminal sequences were different from one another and demonstrated no sequence microheterogeneity. Comparison of these sequences with those in the National Biomedical Research Foundation data bank of protein sequences revealed that the subunits of human myeloperoxidase were not homologous to any known protein. Myeloperoxidase purified from HL-60 cells grown in culture demonstrated the same alpha 2 beta 2 subunit structure. Three isoenzymes of myeloperoxidase, prepared by gradient elution from a CM-Sepharose column, underwent quantitative analysis. No structural basis for the different elution pattern of the myeloperoxidase isoenzymes was discerned by amino-acid analysis, N-terminal sequence, polyacrylamide gel electrophoresis, or digestion with neuraminidase or enzymes known to cleave N-linked heterosaccharides. The structural basis for the myeloperoxidase isoenzymes of human neutrophils, each possessing equivalent activity, is not apparent from these studies. PMID- 3040107 TI - A distinction in vitro between rat liver phosphatidate phosphatase and phospholipase C. AB - Hepatocellular membranes (1000 X g) containing membrane-associated, labeled phosphatidic acid were incubated (1-30 min) with 2 mM oleate or 5 mM bromobenzene in the presence or absence of various metals and NaF. Under the appropriate incubation conditions, membranes displayed rapid and significant oleate- and bromobenzene-dependent increases in the dephosphorylation of labeled phosphatidic acid. However, oleate and bromobenzene activated the dephosphorylation of phosphatidate by phosphatidate phosphatase and phospholipase C, respectively. This conclusion is supported by the observation that the phosphatase stimulated by oleate is: (1) Mg2+ -dependent; (2) inhibited by other metals, such as Ca2+; (3) inhibited by NaF; (4) specific for phosphatidic acid; and (5) associated with a rise in liver cell triacylglycerol production. Bromobenzene, however, activated a phospholipase C that is: (1) stimulated by various metals, such as Mg2+, Ca2+ and Ba2+; (2) insensitive to NaF; (3) associated with the degradation of various membrane phospholipids; (4), associated with liver cell injury; and (5) not associated with a rise in liver cell triacylglycerol formation. These results suggest that under appropriate conditions in vitro the dephosphorylation of phosphatidic acid can be used to assess changes in phosphatidate phosphatase and/or phospholipase C activity. The distinction between these enzymes is important, since phosphatidate phosphatase and phospholipase C regulate key steps in phospholipid biosynthesis and degradation, respectively. PMID- 3040108 TI - Influence of proteinase inhibitors and substrates on 2,3,7,8-tetrachlorodibenzo-p dioxin (TCDD)-binding capacity of the rat hepatic Ah receptor. AB - These studies investigated the effects of various serine proteinase inhibitors and substrates on the TCDD-binding capacity of the rat hepatic Ah receptor. TCDD binding to the Ah receptor was inhibited by serine proteinase inhibitors phenylmethylsulfonyl fluoride (PMSF), tosyl-lysine chloromethyl ketone (TosLysCH2Cl), tosylamide-phenylethyl chloromethyl ketone (TosPheCH2Cl) and substrates tosyl-L-arginine methyl ester (TosArgOMe) and D-tryptophan methyl ester (TrpOMe). The order of potency was TosPheCH2Cl greater than TosLysCH2Cl much greater than PMSF approximately equal to TosArgOMe approximately equal to TrpOMe. Reactivity of the chloromethyl ketones with sulfhydryl groups was suggested by their steep inhibition curves above the concentration of nonprotein sulfhydryl groups, and the partial mitigation of inhibition by 1 mM dithiothreitol. Inhibition by these reagents was irreversible, while that by TosArgOMe and TrpOMe was completely reversible by gel filtration. The mechanism of inhibition by TosArgOMe and TrpOMe was formally competitive, with inhibition constants similar to those reported in steroid hormone receptor systems. Neither inhibitors nor substrates displaced previously bound TCDD. PMID- 3040110 TI - Evidence for non-vacuolar proteolytic catabolite inactivation of yeast fructose 1,6-bisphosphatase. AB - Immunoblotting was used to study whether proteolytic degradation of fructose-1,6 bisphosphatase (EC 3.1.3.11) in yeast cells during catabolite inactivation occurs intra- or extravacuolarly. The 40-kDa subunits of both the phosphorylated and the non-phosphorylated fructose-1,6-bisphosphatase are rapidly degraded by an extract from isolated vacuoles to a 32-kDa intermediate which accumulates and is then slowly further degraded. However, in intact cells, neither the 32-kDa nor any other intermediate reacting with the fructose-1,6-bisphosphatase antibodies is observed following glucose-induced degradation of the enzyme. These observations are discussed as evidence against intravacuolar degradation of fructose-1,6 bisphosphatase during proteolytic catabolite inactivation. PMID- 3040109 TI - The reduction of anti-tumour diaziridinyl benzoquinones. AB - The properties of the semiquinone radicals produced for 2,5-bis(carboethoxyamino) 3,6-diaziridinyl-1,4-benzoquinone (AZQ) and 2,5-bis(2-hydroxyethylamino)-3,6 diaziridinyl-1,4-benzoquinone (BZQ), have been investigated. AZQ semiquinone radicals can be produced from the reduction of AZQ by superoxide radicals, whereas BZQ semiquinone radicals are unstable in the presence of oxygen. The one electron reduction potentials of the couples Q/Q-. at pH 7.0 were determined as 70 +/- 10 mV for AZQ and -376 +/- 15 mV for BZQ. The difference in these potentials is explained. As a consequence of ESR studies on the enzymatically produced radicals, we have considered the factors which determine the detection of ESR signals for reduced quinones produced in a biological system. PMID- 3040111 TI - The reactivity of various free radicals with hyaluronic acid: steady-state and pulse radiolysis studies. AB - The reactions of the primary water radicals with the biopolymer hyaluronic acid have been studied by pulse radiolysis. Bimolecular rate constants, expressed in terms of the disaccharide repeating sub-unit of hyaluronic acid, for OH., H. and eaq- were found to be 7 X 10(8) M-1 X s-1, 5 X 10(7) M-1 X s-1 and less than 5 X 10(6) M-1 X s-1, respectively. By comparing the viscosities of samples, gamma irradiated in the steady state under a variety of conditions, with unirradiated controls, the efficiencies with which selected radicals cause chain breakage have been determined. Efficiencies of 30%, 15%, 0%, 0.2% and 5% were estimated for OH., H., eaq-, methanol radicals and tert-butanol radicals, respectively. The presence of oxygen during irradiation increased the extent of chain breakage by a factor of 1.75. PMID- 3040112 TI - Regulation of fructose 2,6-bisphosphate levels in Neurospora crassa. AB - Both wild type and cr-1 mutant (adenylate cyclase and cyclic AMP-deficient) strains of Neurospora crassa contain fructose 2,6-bisphosphate at levels of 27 nmol/g dry tissue weight. This level decreases by about 50% in both strains upon depriving the cells of carbon or nitrogen sources for 3 h. An increase in cyclic AMP levels produced by addition of lysine to nitrogen-starved cells produced no increase in fructose 2,6-bisphosphate levels. Both strains respond to short-term addition of salicylate, acetate, or 2,4-dinitrophenol with an increase in fructose 2,6-bisphosphate. Thus, the above-described regulation of fructose 2,6 bisphosphate levels is cyclic AMP-independent. A suspension of the wild type produces a transient increase of fructose 2,6-bisphosphate in response to administration of glucose, whereas the mutant strain does not respond unless it is fed exogenous cyclic AMP. Substitution of acetate for sucrose as a sole carbon source for growth leads to a differential decrease in fructose 2,6-bisphosphate levels between the two strains: the wild type strain has 63% and the cr-1 mutant strain has 37% of the levels of fructose 2,6-bisphosphate on acetate as compared to sucrose-grown controls. This may be the basis for an advantage of cr-1 over wild type in growth on acetate. Thus, although most regulation of fructose 2,6 bisphosphate is cyclic AMP-independent, the levels can be regulated by a combination of carbon source and cyclic AMP levels. PMID- 3040113 TI - Effect of administration of diethylhexyl phthalate on the function and turnover of rat hepatic mitochondria. AB - Administration of the widely used plasticizer di(2-ethylhexyl)phthalate (2% w/w) in the diet to the rat caused proliferation of mitochondria in the liver. The number of mitochondria as well as the amount of protein recovered in the organellar fraction was doubled. Mitochondria isolated from the livers of treated animals showed decreased (50%) respiratory activity. The content and activity of cytochrome oxidase were also decreased. The specific incorporation of amino acids into the proteins of whole liver and of mitochondria was not increased in plasticizer-treated animals. Isolated mitochondria also did not show any difference in the rate of incorporation of amino acids into proteins. The half lives of whole liver proteins and of mitochondria were increased in plasticizer fed animals. The half-life of cytochrome oxidase, however, was unaffected by the treatment. The pattern of double labeling of mitochondrial proteins confirmed decreased turnover in plasticizer-treated animals. PMID- 3040114 TI - Cerebroside sulfuric ester (sulfatide) induces oxygen radical generation in guinea-pig leukocytes. AB - Previous studies on experimental allergic encephalomyelitis have shown that a number of leukocytes appear in demyelinating lesions of guinea-pig brain. The present studies showed that cerebroside sulfuric ester (sulfatide), a typical component of myelin membranes, stimulated the oxidative metabolism of guinea-pig neutrophils and macrophages, leading to marked generation of oxygen radicals and light emission. Formation of a spin adduct of 5,5-dimethyl-1-pyrroline N-oxide by leukocytes was dependent on the concentration of sulfatide, and correlated well with the generation of superoxide anion and the intensity of chemiluminescence measured in the absence of luminol. The addition of myelin membranes to the sulfatide-stimulated neutrophils amplified the light emission, suggesting an interaction between myelin membranes and those of leukocytes. Assay of the thiobarbituric acid reaction in the mixture of membranes and cells showed that sulfatide-stimulated cells induced lipid peroxidation in myelin membranes. These results suggest that sulfatide released from demyelinating lesions stimulates leukocytes to release toxic oxygen radicals, which attack myelin membranes, leading to a chain reaction of demyelination. PMID- 3040115 TI - Inhibition of glucagon-stimulated cAMP accumulation and fatty acid oxidation by E series prostaglandins in isolated rat hepatocytes. AB - E-series prostaglandins have been shown to inhibit hepatic glucagon-stimulated glycogenolysis without inhibiting glycogenolysis stimulated by cAMP analogs. In the present studies, prostaglandin E2 and 16,16-dimethylprostaglandin E2 inhibited glucagon-stimulated cAMP accumulation in isolated rat hepatocytes by 25% and 46%, respectively, without affecting basal cAMP levels. Half-maximal inhibition of glucagon-stimulated cAMP accumulation occurred at approx. 10(-7) M 16,16-dimethylprostaglandin E2. 16,16-Dimethylprostaglandin E2 inhibited glucagon stimulated palmitate oxidation in intact hepatocytes without affecting basal rates of palmitate oxidation. 16,16-Dimethylprostaglandin E2 had no effect on palmitate oxidation in a liver homogenate system. These studies demonstrate that prostaglandin E antagonizes the effects of glucagon on hepatic metabolism by inhibiting glucagon-stimulated cAMP accumulation. PMID- 3040116 TI - Difference in extracellular radical release after chemotactic factor and calcium ionophore activation of the oxygen radical-generating system in human neutrophils. AB - Results obtained with the luminol-dependent chemiluminescence technique show that with this technique, generation of radicals from an extra- as well as from an intracellular source is quantified. By means of a chemiluminescence technique, using human neutrophils stimulated with the chemoattractant formylmethionylleucylphenylalanine and the calcium ionophore ionomycin, two different mechanisms of radical production and release are demonstrated. The chemoattractant causes the cells to produce oxygen radicals which to a large extent are released from the cells. The calcium ionophore is also capable of stimulating radical formation but does not suffice for extracellular release. Furthermore, the removal of extracellular Ca2+ is of minor importance for the extracellular radical production, whereas it totally inhibits the generation of radicals with an intracellular localization. The mechanism(s) behind intracellular and extracellular production of oxygen radicals is discussed. PMID- 3040117 TI - Two subpopulations of alpha 1-adrenergic receptors with high and low affinity for agonists: short-term exposure to agonists reduced the high-affinity sites. AB - Short-term receptor regulation by agonists is a well-known phenomenon for a number of receptors, including beta-adrenergic receptors, and has been associated with receptor changes revealed by radioligand binding. In the present study, we investigated the rapid changes in alpha 1-adrenergic receptors induced by agonists. alpha 1-receptors were studied on DDT1 MF-2 smooth muscle cells (DDT1 MF-2 cells) by specific [3H]prazosin binding. In competition binding on membranes and on intact cells at 4 degrees C or at 37 degrees C in 1-min assays, agonists competed for a single class of sites with relatively high affinity. By contrast, in equilibrium binding at 37 degrees C on intact cells agonists competed with two receptor forms (high- and low-affinity). We quantified the receptors in the high affinity form by measuring the [3H]prazosin binding inhibited by 20 microM norepinephrine (this concentration selectively saturated the high-affinity sites). The low-affinity sites were measured by subtracting the binding of [3H]prazosin to the high-affinity sites from the total specific binding. High affinity receptors were 85% of the total sites in binding experiments at 4 degrees C, but only 30% at 37 degrees C. On DDT1-MF-2 cells preequilibrated with [3H]prazosin at 4 degrees C, and then shifted to 37 degrees C for a few minutes, norepinephrine selectively reduced the high-affinity sites by 30%. We suggest that at 4 degrees C it is the native form of alpha 1-receptors that is measured, with most of the sites in the high-affinity form, while during incubation at 37 degrees C the norepinephrine present in the binding assay converts most of the receptors to an apparent low-affinity form, so that they are no longer recognized by 20 microM norepinephrine. The nature of this low-affinity form was further investigated. On DDT1-MF-2 cells preincubated with the agonist and then extensively washed at 4 degrees C (to maintain the receptor changes induced by the agonist) the number of receptors recognized by [3H]prazosin at 4 degrees C was reduced by 38%. After fragmentation of the cells, the number of receptors measured at 4 degrees C was the same in control and norepinephrine-treated cells, suggesting that the disruption of cellular integrity might expose the receptors which are probably sequestered after agonist treatment. In conclusion, the appearance of the low affinity for agonists at 37 degrees C may be due to the agonist-induced sequestration of alpha 1-adrenergic receptors, resulting in a limited accessibility to hydrophilic ligands. PMID- 3040118 TI - Polyamines stimulate superoxide production in human neutrophils activated by N fMet-Leu-Phe but not by phorbol myristate acetate. AB - The polyamines putrescine, spermidine and spermine, at concentrations of 10 microM, stimulated superoxide generation by human polymorphonuclear leukocytes induced by fMet-Leu-Phe in the presence of Ca2+. This positive effect was not evident in the absence of Ca2+ or when the polymorphonuclear leukocytes were stimulated by phorbol myristate acetate. Spermidine in the range of 10-100 microM showed a dose-dependent stimulatory effect on the superoxide generation induced by fMet-Leu-Phe, whilst at doses above 25 mM it produced an inhibitory effect. At this concentration, spermidine did not reduce the phorbol myristate acetate neutrophil-induced O2-. generation, while an inhibitory effect by the polyamine was evident at concentrations above 50 mM. In addition, 100 microM spermidine increased the amount of superoxide generated and enhanced the ability of the chemotactic peptide to stimulate superoxide generation. The polyamines in the range of 10 microM-25 mM did not modify the activity of purified NADPH oxidase, nor the rate of reduction of cytochrome c as supported by the xanthine/xanthine oxidase reaction. These results indicate that physiological concentrations of polyamines can stimulate superoxide formation by polymorphonuclear leukocyte cells produced by the chemotactic peptide fMet-Leu-Phe, probably by increasing the availability of external calcium. PMID- 3040120 TI - The effect of vitamin D status on cutaneous sterologenesis in vivo and in vitro. AB - Recent studies have shown that cutaneous sterologenesis is autonomous from the influence of circulating sterols, and that the epidermis is an important site of sterologenesis which is regulated by permeability barrier requirements. In addition to barrier function, an additional, important function of the epidermis is to synthesize sterol precursors of vitamin D3. The present study was designed, first, to determine whether vitamin D status and/or circulating levels of 1,25 dihydroxyvitamin D3 might play a role in regulating cutaneous sterol synthesis in vivo and, second, whether 1,25-dihydroxyvitamin D3 modulates sterologenesis in cultured human keratinocytes. Hairless mice were maintained on a vitamin D deficient diet in the dark and supplemented with various doses of vitamin D3/day. Despite demonstrating serum 25-hydroxyvitamin D3 levels ranging from less than 10 to 343 ng/ml, the incorporation of tritiated water into cholesterol and total nonsaponifiable lipids in both the epidermis and dermis was similar in the four groups of animals. Likewise, administration of various doses of 1,25 dihydroxyvitamin D3 to vitamin D-deficient mice resulted in serum levels of 1,25 dihydroxyvitamin D3 ranging from less than 10 to 85 pg/ml; yet, cholesterol and total nonsaponifiable lipid synthesis was similar in both the dermis and epidermis in all groups of animals. Moreover, administration of 0.6 micrograms/kg per day of 1,25-dihydroxyvitamin D3 to 'normal' vitamin D-replete mice also had no effect on cutaneous sterol synthesis. Furthermore, conversion of 7 dehydrocholesterol to cholesterol in vitamin D-deficient vs. supplemented animals did not differ. Finally, addition of 1,25-dihydroxyvitamin D3 to cultured keratinocytes over a concentration range of 10(-12)-10(-7) M did not affect sterologenesis, except at supraphysiologic doses (10(-7) M). Together, these results suggest that vitamin D status does not influence sterol synthesis in the skin. PMID- 3040119 TI - Polarity of the Forssman glycolipid in MDCK epithelial cells. AB - To determine whether epithelial plasma membrane glycolipids are polarized in a manner analogous to membrane proteins, MDCK cells grown on permeable filters were analyzed for the expression of Forssman ceramide pentasaccharide, the major neutral glycolipid in these cells. In contrast to a recent report which described exclusive apical localization of the Forssman glycolipid (Hansson, G.C., Simons, K. and Van Meer, G. (1986) EMBO J. 5, 483-489), immunofluorescence and immunoelectron microscopic staining revealed the Forssman glycolipid on both the apical and basolateral surfaces of polarized cells. Immunoblots indicated that the Forssman antigen was detectable only on glycolipids and not on proteins. Analysis of metabolically labeled glycolipids released into the apical and basal culture medium, either as shed membrane vesicles or in budding viruses, also demonstrated the presence of the Forssman glycolipid on both apical and basolateral membranes of polarized cells. Quantitation of the released glycolipid indicated that the Forssman glycolipid was concentrated in the apical membrane. These results are consistent with previous reports which described quantitative enrichment of glycolipids in the apical domain of several epithelia. PMID- 3040121 TI - Physical characterization of native opiate receptors. Additional information from detailed binding analysis of a radiation-inactivated receptor. AB - Target size analyses of the etorphine receptor were performed on frozen rat brain P2 homogenates using the radiation inactivation technique. Multi-point saturation curves at each radiation dose revealed that the apparent dissociation constant for the binding of this ligand to its receptor is a function of the dose. Analysis of the results shows clearly that the ligand-binding macromolecule is functionally coupled to at least one other macromolecule. When the coupling is destroyed the ligand dissociation constant becomes larger by over an order of magnitude. Thus, the variation of the dissociation constant with dose yields interesting new information on the nature of the native receptor which has implications with respect to the conformation of the binding site and to solubilization and cloning methods directed towards sequencing the ligand-binding component of opiate receptors. PMID- 3040122 TI - Protein phosphorylation associated with stimulation of rabbit gastric glands. AB - Changes in protein phosphorylation associated with stimulation of acid secretion were investigated using isolated rabbit gastric glands labeled with 32P. The glands were stimulated by 100 microM histamine plus either 10 microM forskolin or 50 microM isobutylmethylxanthine, homogenized, and fractionated into a series of pellets: 40 X g, 5 min; 4000 X g, 10 min; 14,500 X g, 10 min; 48,200 X g, 90 min (microsomes); and supernatant. Stimulation induced a redistribution of H+/K+ transporting ATPase among the membrane fractions, i.e., a reduction in activity of the microsomal fraction, and a compensatory increase in the 4000 X g fraction. Further subfractionation of the 4000 X g pellet by Ficoll density gradient produced an 18% Ficoll layer, greatly enriched in the H+/K+-ATPase, and which is thought to be rich in apical membranes of parietal cells. SDS-polyacrylamide gel electrophoresis showed that the amount of 94 kDa peptide (the molecular size of the H+/K+-ATPase) was increased in the 18% Ficoll layer and decreased in the microsomal fraction by stimulation. Analysis of autoradiograms of the gels revealed that apparent changes in level of phosphorylation occurred in the 120, 94 and 80 kDa regions of the 18% Ficoll layer, and in the 94 kDa region of the microsomal fraction. The phosphorylation changes in the 94 kDa region may not reflect changes in specific activity of a single peptide but may be due to the heterogeneity of proteins in this region, which was demonstrated by selective heat treatment of the samples as well as two-dimensional electrophoresis. Phosphorylation of 120 kDa protein in the 18% Ficoll layer was clearly increased by stimulation, and this appeared to be associated with protein distribution changes as well as phosphorylation. The 80 kDa protein in the 18% Ficoll layer showed marked increased phosphorylation by stimulation, with little change in protein distribution. This 80 kDa protein was focused on two-dimensional gels as several sequential spots, with the most radioactive peptide focused toward the acidic side; thus, we propose isomeric forms of an 80 kDa protein with sequential phosphorylation sites. The phosphorylation changes observed in this study are considered to be important to the process of gastric acid secretion because they occurred in the putative apical membrane fractions in which biochemical and functional changes with stimulation have been demonstrated. PMID- 3040123 TI - Identification of a phosphorylated form of phosphoenolpyruvate carboxykinase from the yeast Saccharomyces cerevisiae. AB - A phosphoprotein of 65 kDa, as determined by SDS-gel electrophoresis, has been isolated from yeast crude extracts. This phospho form copurifies with phosphoenolpyruvate carboxykinase in the enzyme purification procedure worked out in our laboratory (Tortora, P., Hanozet, G.M. and Guerritore, A. (1985) Anal. Biochem. 144, 179-185). Moreover, both proteins bind strongly to 5'AMP-Sepharose 4B in the presence of Mn2+, whereas a substantially lower binding occurs if Mn2+ is replaced by Mg2+. This binding pattern is consistent with the well-known Mn2+ dependence of yeast phosphoenolpyruvate carboxykinase. These data suggest that the 65-kDa protein might be a phosphorylation product of the native enzyme. Furthermore, although the phospho form is not immunoprecipitated by anti phosphoenolpyruvate carboxykinase antibodies, addition of Protein A-Sepharose CL 4B to crude extracts preincubated with the antibodies results in the binding to the resin of the phospho form, thus providing immunological evidence for its identification as a modified form of native enzyme. The same 65-kDa phosphoprotein is detectable in extracts from cells grown in the presence of [32P]Pi, as well as in cell extracts incubated with [gamma-32P]ATP. Moreover, digestion of the phosphoprotein with BrCN or with Staphylococcus aureus V8 proteinase, yields two and three fragments, respectively, which appear parallel to digestion products of phosphoenolpyruvate carboxykinase, again supporting the proposed identification. Finally, analysis of the phosphorylated amino acids in the 65-kDa protein shows that phosphoserine is the only labelled phosphoamino acid. PMID- 3040124 TI - Inhibition of receptor-mediated stimulation of cyclic AMP accumulation in neuroblastoma-hybrid NCB-20 cells by a phorbol ester. AB - Activation of protein kinase C by phorbol esters such as phorbol 12-myristate 13 acetate (PMA), modulates responsiveness of the cyclase system in many cell types. In the neuroblastoma-hybrid cell line NCB-20, PMA causes a reduction in receptor mediated accumulation of cyclic AMP. The reduction in receptor responses by PMA occurs within 3 min and is still apparent at 40 min. This occurs in a concentration-dependent manner with an EC50 for PMA of approx. 30 nM. Accumulations of cyclic AMP that are elicited by prostaglandin E2, vasoactive intestinal peptide or 2-chloroadenosine are decreased in the presence of PMA. Accumulations of cyclic AMP that are elicited by forskolin in the absence of a receptor agonist are unaffected by the presence of PMA. Inhibition of cyclic AMP generation by dopamine is not diminished by PMA suggesting the receptor input through the inhibitory Ni-guanyl nucleotide binding protein is still functional after PMA treatment. The generalized inhibition of receptor-mediated responses by PMA could be due to a protein kinase C-mediated phosphorylation of the stimulatory Ns-guanyl nucleotide binding protein, but other mechanisms are possible. PMID- 3040125 TI - Primary structure analysis proves that protein phosphatases 2C1 and 2C2 are isozymes. AB - The two recently discovered forms of protein phosphatase 2C, termed 2C1 and 2C2, were digested with CNBr or trypsin, and several peptides corresponding to two regions of the protein were sequenced. These studies revealed close homology between the two enzymes with 49 identities over the 62 residues that could be compared directly. The results establish that protein phosphatases 2C1 and 2C2 are the products of different genes. The C-terminus of protein phosphatase 2C2 has also been identified. PMID- 3040126 TI - [Effect of temperature and binding with copper ions on the motility of spin labeled oxy- and methemoglobin subunits]. AB - Using the method for separate determination of correlation times of spin-labeled proteins (tau M) and labels (tau R) it has been shown that at temperatures below and about 25 degrees the mobility of oxy Hb subunits is higher than that of met Hb. From 30 degrees on oxy and met Hb show identical flexibility (tau M = 40 ns) in 0.01 M phosphate buffer (pH 7.3) containing 0.15 M NaCl. With a decline in pH from 7.3 to 6.4 the intramolecular mobility of met Hb subunits decreases. In the absence of 0.15 M NaCl at pH 7.3 (5 degrees C) met Hb becomes more flexible (tau M drops from 25 to 16 ns). Complex formation of beta-chains of oxy Hb with one Cu+2 ion at 20 degrees has a negligible bearing on the flexibility of the protein, whereas addition of second ion considerably enhances interaction between the subunits of the tetramer and decreases its flexibility (tau M rises from 17 to 30 ns). PMID- 3040127 TI - [Inhibitory effect of active oxygen scavengers on the endocytosis of concanavalin A by lymphocytes]. AB - It has been shown that of active oxygen species regulate the activity of the processes of Con A endocytosis in rat thymus cells. PMID- 3040128 TI - [Structural state of the cardiomyocyte sarcolemma in animals of different ages]. AB - Cardiomyocyte plasmalemma membranes of adult (12 months) and old (24-26 months) rats have been studied. Ageing is accompanied with an essential decrease of the content of the membrane protein SH-groups. We have found an increase in the fraction of strongly immobilized spin-labeled SH-groups, a decrease in that of unsaturated lipids, as well as an increase in that of lysophosphatidylcholine in old rats membranes. The pattern of the response of beta-adrenergic receptors to isoproterenol is changed with age. The mobility of spin probe 5NS in the membranes of old rats is lowered at 22.5 degrees C. However there is no statistically significant age difference in the spin-probe mobility at 37 degrees C. Possible role of lysophosphatidylcholine in the stabilization of membrane lipid "fluidity" at physiological temperatures is suggested. PMID- 3040129 TI - [Changes in the segmental flexibility and conformation stability of side groups in spin-labeled molecules of 2 IgG subclasses in the serum of normal cows and cows in various pathological states]. AB - The segmental flexibility of molecules from two IgG subclasses of cows' blood serum was first observed. It was shown that IgG1 molecule has a more flexible hinge region than IgG2 molecule and lower local conformational stability. At the same time the states of IgG1 and IgG2 molecules determined by dynamic-steric characteristics for ill and healthy cows practically did not differ. PMID- 3040130 TI - [Effect of oxidative phosphorylation inhibitors and uncoupling agents on cAMP activity]. AB - Uncouplers of oxidative phosphorylation increased the speed of substrate oxidation and ATP hydrolysis and raised cAMP induced neuron membrane current. Different inhibitors decreased it. Both effects support the hypothesis that a signal of intracellular injected cAMP spreads to the neuron membrane as a mechanical signal. This signal propagated to the membrane along microtubules which according to this hypothesis serve as a sound generator with metabolic heat pumping. PMID- 3040131 TI - [Effect of elevated concentrations of sodium chloride and temperature on the state of erythrocyte cytosol]. AB - It was shown by means of ESR-spin probe method that microviscosity of erythrocyte cytosol had a limiting value which could be reached by various concentrations of sodium chloride depending on temperature. With a temperature decrease when reaching this value haemolysis occurs. PMID- 3040132 TI - [Effect of the opiate antagonist naloxone on the electrical activity of identified neurons in the edible snail]. AB - The opiate antagonist naloxone modifies the electric activity of some identified neurons of the Helix lucorum which have not been preliminary exposed to the effect of exogenous opioids. Some neurons are excited while others are inhibited by naloxone, and in both cases the reaction may have both a short and long latent period. The reactions depend on naloxone dose and become less expressed or are blocked when naloxone is administered together with the agonists of opiate receptor (morphine, D-Ala2, D-Leu5-enkephalin, bremazocine and beta-endorphin). Opioids alone do not produce any effect on neurons. The effect of naloxone on neurons is assumed to be a result of the elimination by the opiate antagonist of the tonic effect of endogenous opioids by their replacing on opiate receptors which are constantly stimulated by endogenous ligands. PMID- 3040133 TI - [Transmission of amplitude-modulated signals in the presynaptic inhibition system of the cat spinal cord]. AB - In experiments with cats under the hexenal anesthesia it has been found that the transmission of information in multineuronic reflex ring carries out by frequency dependent way. The optimum frequency subrange corresponds to each meaning of the modulation depth. Appearing signal distortions are accompanied by the phase lead of dorsal potentials with maximum on "resonance" frequency. The mechanism of presynaptic inhibition serves as control command amplifier. For all this the information signal amplitude grows, signal relation to noise increases, noise immunity of information transmission through the neuronic communication channel of multineuronic reflex ring improves considerably. PMID- 3040135 TI - Human umbilical arteries and veins: generation of leukotrienes and response to exogenous leukotrienes. AB - When human umbilical arteries and veins were challenged with calcium ionophore A23187 they released a leukotriene (LT)-like material. On bioassay using guinea pig ileal smooth muscle, the release was equivalent to 37.4 +/- 7.5 ng LTD4 (mean +/- SEM) per gram of arterial tissue, and 29.0 +/- 4.7 ng LTD4/g of venous tissue. The LT was further purified by high pressure liquid chromatography, and characterised as LTC4 by bioassay and radioimmunoassay of purified fractions. Longitudinal strips from human umbilical arteries in vitro did not contract to doses of synthetic LTD4 in the range 5 X 10(-11) to 10(-9) mol. PMID- 3040134 TI - Development of enzymes of glycerol metabolism in human fetal liver. AB - The activities of three key enzymes of glycerol metabolism were measured in liver samples from 37 human fetuses ranging in gestational age from 18 weeks to term, from neonates (1-3 days) and from infants to 2 years. Glycerol kinase specific activity was constant throughout the period of fetal development examined, and was comparable to that measured in neonates and infants. However, the subcellular distribution of the activity changed markedly, being predominantly particulate in fetal samples and cytoplasmic in postnatal samples. The particulate activity had an elevated Km for glycerol. Cytoplasmic glycerol-3-phosphate dehydrogenase activity was very low in the fetal period, and then rose to adult levels during infancy. There were no kinetic differences between the fetal and postnatal activities. Mitochondrial glycerol-3-phosphate dehydrogenase activity rose somewhat after birth to near adult levels. The data indicate that glycerol can be metabolized by human fetal, neonatal and infant liver. PMID- 3040136 TI - Steroid-induced final maturation in brook trout (Salvelinus fontinalis) oocytes in vitro: the effects of forskolin and phosphodiesterase inhibitors. AB - The effects of phosphodiesterase inhibitors and forskolin on steroid-induced germinal vesicle breakdown (GVBD) were investigated in brook trout (Salvelinus fontinalis) oocytes using an in vitro incubation technique. Follicles were first treated with a collagenase solution to remove the follicle wall. Denuded oocytes were examined, using scanning electron microscopy. In all experiments GVBD was induced by the use of 17 alpha, 20 beta-dihydroxy-4-pregnen-3-one. Cyclic adenosine 3',5'-monophosphate (cAMP) levels were measured (by protein-binding assay) in control and forskolin-treated oocytes. Collagenase treatment removed a majority of the follicle wall, as shown by scanning electron microscopy. Partially denuded (PD) oocytes were slightly more sensitive to steroid treatment than intact follicles (IF), as shown by ED50 values; but PD oocytes did not respond to gonadotropin (GTH) stimulation. Both 3-isobutyl-1-methyl-xanthine (IBMX) and SQ20,006 (Squibb) blocked GVBD, but IBMX was more inhibitory. Forskolin also blocked steroid-induced GVBD. Kinetics of inhibition studies were performed using IBMX, forskolin, and cycloheximide. IBMX and cycloheximide inhibited GVBD if added during the first 18 h following steroid stimulation, whereas forskolin blocked GVBD if added within 12 h after steroid treatment. Forskolin, at levels that block GVBD in vitro, significantly increased cAMP in both IF and PD oocytes, but the response of IF was greater than that of PD oocytes. PMID- 3040137 TI - Coxsackie B myocarditis. PMID- 3040138 TI - The determination of alpha-adrenergic receptor concentration on rat pancreatic islet cells. AB - The selective alpha 2 adrenergic antagonist yohimbine has been shown to prevent the noradrenaline induced inhibition of insulin secretion from isolated rat islets of Langerhans. Binding studies utilizing [3H]yohimbine showed specific binding to dispersed rat islet cells with a Kd of 2.9 nM and receptor concentration of 645 fmols/mg protein. The use of chloroquine to inhibit receptor recycling did not affect binding of the ligand. Binding studies and secretion data are consistent with the suggestion that adrenergic receptors of the alpha 2 sub-type may play a dominant role in the regulation of insulin secretion. PMID- 3040139 TI - Hormone evolution studies: multiplication promoting and imprinting ("memory") effects of various amino acids on Tetrahymena. AB - Aromatic, heterocyclic, polar and non-polar amino acids were examined for imprinting potential in a unicellular (Tetrahymena) model system. Serine gave rise to positive, glycine to negative imprinting, whereas valine, tryptophan, tyrosine and phenylalanine had no imprinting effect whatever. However, tyrosine and phenylalanine stimulated the division of Tetrahymena already at primary interaction, the former even for a relatively long time. It follows that amino acids, too, can give rise to imprinting, although their imprinting potentials are dissimilar. These phenomena have attracted attention to possible interrelationships between the supposed amino acid receptors of Tetrahymena and the evolution of amino acids to hormones. PMID- 3040140 TI - Overlapping imprinting of oligopeptides in Chang liver cells. Data on the mechanism of hormone evolution. AB - Imprinting was induced with synthetic oligopeptides in Chang liver cell cultures to test these molecules for signal molecule value. Investigations into imprinting overlaps (cross-imprinting) have shown that all oligopeptides (di-, tetra- and pentapeptides) carrying a terminal proline group were able to imprint the cells for the pentapeptide Tyr-D-Met-Gly-Phe-Pro-NH2, which displayed an outstanding imprinting potential for itself and an extraordinary opioid activity as well. The fact that exclusively the proline-deficient oligopeptide (a tetrapeptide) failed to imprint for the pentapeptide in question, indicates a decisive role of proline in the transformation of molecules to signal carriers (hormones). The pentapeptide in question did imprint for the related molecules (except the dipeptide) but to a much lesser degree than for itself. The marked inferiority of the pentapeptide's cross-imprinting potential to its self-imprinting potential supports the hypothetical implication that a considerable difference between the specific and non-specific binding capacities of a molecule, if not the loss of non-specific binding was an essential prerequisite of transformation to a signal molecule, i.e. of hormone evolution. PMID- 3040141 TI - New neuromuscular symptoms after old polio ("the post-polio syndrome"): clinical studies and pathogenetic mechanisms. PMID- 3040142 TI - [Adrenal glucocorticoid function and its regulation in human prenatal ontogeny and in the 1st week of life]. AB - A comparison of cortisol and ACTH level determination in 82 human fetal sera obtained in the 11th-34th week of the intrauterine development and 50 sera from infants 1-7 days of age has revealed several stages of pituitary-adrenal system (PAS) maturation. PAS is immature in the first trimester of embryogenesis when cortisol concentration in the human fetal blood is low. The functional relations between pituitary and adrenal glands begin to establish in the middle of prenatal developmental period. A prominent response to delivery stress indicates that by delivery PAS reaches a certain stage of maturation. The feedback between ACTH and cortisol secretions in the blood of infants 1-7 days of age demonstrates that the refractory period of PAS is absent in the neonates during the first week of their life. PMID- 3040143 TI - [Age-related and regional changes in angiotensin-converting and kinin-degrading activity in the brain of aggressive rats]. AB - Aggressive behaviour (muricidal) induced in rats by local electrolyte lesions of septal brain area was accompanied by changes in angiotensin-converting enzyme (ACE) and total kinin-destroying activity (KDA) in different brain regions. In 2 month-old rats (30 days after septal operation) a decrease in ACE activity was observed in hypothalamus and striatum, while in the cerebellum the activity was increased. KDA in this group of aggressive rats was markedly increased in the pituitary body, hypothalamus and striatum. Half a year after septal lesion in spite of aggressive behaviour retention, ACE activity and KDA did not differ from their activity in nonmuricidal rats of the same age. These data show a distinct role of the peptides studied in the formation and maintenance of muricidal behaviour in rats of different age. PMID- 3040144 TI - [Changes in the activity and regulatory properties of Na, K-ATPase from the myocardial sarcolemma in total graded ischemia]. AB - Na, K-ATPase activity of the rat and guinea-pig myocardial sarcolemma and its sensitivity to digoxin (DG) and carbamylcholine (CCh) were investigated during experimental ischemia. Ischemia was induced by the incubation of hearts in the air at 37 degrees C. This 15-, 30- and 45-min treatment led to a decrease in enzymatic activity which was similar in both animal species. Dose-related dependence of DG effect (10(-8)-10(-2) M) on sarcolemmal Na, K-ATPase activity of guinea-pig ischemic hearts did not differ from the control, whereas the rat enzyme sensitivity to glycosides rose with the progress of ischemia. CCh (10(-7) 10(-3) M) produced an inhibition of Na, K-ATPase activity which had reached 40% both in the rat and guinea-pig myocardial preparations. This effect was blocked by atropine (10(-6) M). The magnitude of enzyme responses to CCh declined depending on the duration of ischemia, with it being greater in guinea-pig sarcolemma than in rat membrane. The increased sensitivity of the rat Na, K ATPase to CCh was also observed. PMID- 3040145 TI - [Anti-arrhythmic action of adenosine and the cyclic nucleotide content of the myocardium of rats with ischemia]. AB - In anesthetized rats a 30-min intravenous infusion of adenosine (2.5 mg/kg/min) performed after the coronary artery ligation significantly decreased the incidence and severity of early ischemic arrhythmias. After the infusion of adenosine, there was an increase in cGMP level in the left ventricular myocardium, cAMP content remained unchanged. PMID- 3040146 TI - [Spontaneous and septal models of muricidal behavior: regional activity of enzymes regulating angiotensin and bradykinin metabolism in the brain]. AB - Spontaneously muricidal rats (SMR) were selected from a group of white male rats. In the remaining animals muricide was induced by the local electrolyte damage of the brain septal area. Both muricidal models had different physiological indexes of aggressive reactions. In SMR a significant decrease in angiotensin-converting enzyme activity was detected in the midbrain and thalamus-hypothalamus areas. In the group of operated muricidal ("septal") rats alterations in angiotensin converting enzyme activity have been revealed in none of the brain areas examined. The increase in total kinin-destroying activity in the pituitary, cerebellum, striatum and thalamus-hypothalamus areas was detected. The results indicate neurochemical specificity of brain angiotensin II and kinins in the regulation of different muricidal models. PMID- 3040147 TI - [Theophylline-induced changes in the presynaptic effects of adenosine, baclofen, clofelin and morphine in the myenteric plexus of the guinea pig]. AB - The experiments on the isolated guinea-pig ileum have shown that the contractions caused by low frequency transmural stimulation are decreased by adenosine, baclofen, clofelin and morphine. Theophylline is a competing antagonist of adenosine and a noncompeting antagonist of baclofen, clofelin and morphine. The effects of the latter are not altered by 1,3-dipropyl-8-phenylxanthine suppressing adenosine effects. It is concluded that presynaptic effects of baclofen, clofelin and morphine depend on cAMP level in cholinergic neurons of myenteric plexus. PMID- 3040148 TI - [Alpha interferon interaction with opiate receptors in the rat brain]. AB - The preferential interactions of alpha-interferon (alpha-IFN) with delta and mu opiate receptors were studied. alpha-IFN (specific antiviral activity 2 X 10(3) U/mg protein) was shown to inhibit in the competitive manner 3H-naloxone and 3H-D ala2, D-leu5-enkephalin (3H-DADL) specific binding to opiate receptor subpopulations. alpha-IFN was much more effective in decreasing 3H-DADL than 3H naloxone binding in opiate receptors: K1 values averaged 160 +/- 30 and 1150 +/- 80 U/ml, respectively. IFN effective concentrations inhibiting 50% of 3H-naloxone opiate receptor binding in the absence or presence of 100 mmol/l NaCl were similar, and the "sodium shift" value was equal to 1. The independence of alpha IFN activity of the presence of NA+ cations suggests the antagonist character of alpha-IFN interaction with opiate receptors. Thus, alpha-IFN employed appears to be an alpha-selective ligand displaying the in vitro properties of "pure" morphine antagonists. PMID- 3040150 TI - [Deliberate heart arrest in man and its possible mechanism]. AB - A case of deliberate 7-second heart arrest is described. It is assumed that the effect is the result of voluntary self-irritation of peritoneal receptors by means of rhythmical diaphragm muscle contractions realized in conditions of preliminary breathing arrest. PMID- 3040149 TI - [Opioid activity of peptides and wound healing of the skin]. AB - The binding of dalargin, its four analogues and FK-33824, DADLE, met-enkephalin and morphine to peripheral mu- and delta-receptors and to brain receptors has been investigated in comparison with their influence on skin wound healing in rats. It has been shown that only substances with opiate activity, including morphine, stimulated wound healing. No correlation between wound healing effect of peptides and their binding to a definite receptor has been found. Naloxone inhibited wound healing and suppressed opiate peptide-mediated healing process. It is suggested that endogenous opiate peptides are involved in the maintenance of structural homeostasis. PMID- 3040151 TI - [Effect of corticotropin and hydrocortisone on the Ca2+-ATPase activity of skeletal muscle sarcoplasmic reticulum]. AB - The effect of corticotropin (ACTH1-39), synacthen (ACTH1-24) and hydrocortisone hemisuccinate on the activity of Ca-ATPase of skeletal muscle sarcoplasmic reticulum (SR) and calcium (Ca) accumulation in SR vesicles has been studied. It has been shown that ACTH1-39 (I U per 100 g body weight) increased the activity of Ca-ATPase in skeletal muscle SR of rats, while hydrocortisone (5 mg per 100 g body weight) did not change the activity of Ca-ATPase in skeletal muscle SR. However, both hormones increase the total activity of ATPase. ACTH1-39 and ACTH1 24 (0.05-0.0005 U/ml) and hydrocortisone (2.8 X 10(-7)-2.8 X 10(-9) mol/l) increased in vitro Ca-ATPase isolated from rabbit skeletal muscle SR and accumulation of Ca is SR vesicles. At the same time, hydrocortisone reduced calcium/phosphorus ratio, while ACTH1-39 and ACTH1-24 increased it, i.e. hydrocortisone facilitated Ca accumulation in SR requiring more ATP energy, whereas ACTH facilitated Ca accumulation in SR requiring less ATP energy. PMID- 3040152 TI - [Interstrain differences in changes in 5-nucleotidase activity in mouse macrophages in response to immunostimulation]. AB - The effect of immunostimulants on the activity of 5-nucleotidase in the macrophages of peritoneal exudate (MPE) has been investigated in mice of various strains. It has been demonstrated that in case of subcutaneous introduction of immunostimulants interstrain differences might be observed in the changes of MPE 5-nucleotidase activity. The decrease in the enzymatic activity in MPE was found to be the most pronounced in C57Bl/6 mice, while in AKR mice it was the least marked. CBA and C3 mice revealed no changes in MPE 5-nucleotidase activity in response to immunostimulation. PMID- 3040153 TI - Exposure to gamma-irradiation increases phorbol myristate acetate-induced H2O2 production in human macrophages. AB - Cell number, protein, and phorbol myristate acetate (PMA)-induced H2O2 production were measured in cultured human peripheral blood monocytes for six days after exposure to varying doses of gamma-radiation. Both the number of adherent cells and the protein per dish decreased with increasing radiation doses. The dose of radiation decreasing the number of adherent cells by 37% on days 4 and 6 postirradiation was 29 Gy. Four hours postirradiation there was a small decrease in PMA-induced H2O2 production for doses of 7.5 Gy or greater; levels returned to normal by eight hours and increased at 24 hours postirradiation. By day 4 postirradiation significant increases in PMA-induced H2O2 production were noted at all radiation doses (2.5 to 50 Gy). This increase was not due to a shift in the PMA dose-response curve, a change in the time course of the PMA response, or an effect of decreased cell density on the assay system. Superoxide levels were not significantly changed in cells exposed to 20 Gy. Catalase, glutathione peroxidase, and superoxide dismutase levels also were unchanged. Culturing irradiated cells with gamma-interferon increased PMA-induced H2O2 release, which indicated that irradiated cells retained their capacity to respond to gamma interferon. These data demonstrate that irradiation affects the PMA-induced H2O2 production of human monocytes in a time- and dose-dependent manner. An increase in the release of reactive oxygen intermediates by the macrophage may play a role in enhancing the deleterious effects of radiation in vivo. PMID- 3040154 TI - ADP-induced platelet shape change and mobilization of cytoplasmic ionized calcium are mediated by distinct binding sites on platelets: 5'-p fluorosulfonylbenzoyladenosine is a weak platelet agonist. AB - Platelet stimulation with ADP results in several responses, including shape change, increase in cytoplasmic ionized calcium concentration [Ca2+]i, an inhibition of adenylate cyclase. 5'-p-Fluorosulphonyl benzoyladenosine (FSBA), which covalently labels an ADP binding site on platelets, blocks platelet shape change but not the inhibition of cyclic AMP levels by ADP, whereas p chloromercuribenzenesulfonate (pCMBS), a nonpenetrating thiol reagent, has the opposite effects. We examined the effect of FSBA and pCMBS on ADP-induced increase in [Ca2+]i using platelets loaded with fluorescent Ca2+ indicators quin2 and fura-2. FSBA (50 to 200 mumol/L) induced a dose-dependent rise in [Ca2+]i, indicating that it is a weak platelet agonist. Under conditions of covalent labeling of the ADP binding sites, FSBA (50 to 100 mumol/L) did not inhibit the ADP-induced increase in [Ca2+]i or its inhibition of adenylate cyclase, whereas pCMBS (up to 1 mmol/L) abolished both these responses but not shape change. These findings suggest that ADP-induced Ca2+ mobilization and inhibition of adenylate cyclase are mediated by platelet binding sites distinct from those mediating shape change. PMID- 3040155 TI - Defensin-rich dense granules of human neutrophils. AB - Defensins are a newly recognized class of small, cationic polypeptides that have in vitro microbicidal activity toward certain bacteria, fungi, and viruses. Human neutrophil granules were separated into 13 density fractions by using a high resolution Percoll gradient centrifugation procedure, and the distribution of the three defensin polypeptides in these fractions was determined. Levels of defensins and several granule marker proteins were estimated in each fraction from relative staining intensities of bands following acid-urea and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of total acid extractable proteins. These results were confirmed by enzyme immunoassay measurements of defensins and quantitative determinations of the typical azurophil granule components, myeloperoxidase, beta-glucuronidase, lysozyme, and elastase. The five higher density granule fractions (H1 through H5) contained fourfold higher relative amounts of defensins as compared with the eight lower density fractions (L1 through L8), accounting for approximately 50% of the total protein. In particular, fraction H5 was especially enriched in defensins but was relatively deficient in myeloperoxidase, beta-glucuronidase, lysozyme, and elastase. Ultrastructural morphology showed that fraction H5 contained the largest granules. Seventy percent of these granules exhibited electron-dense rims and electron-lucent central regions when stained with methanolic uranyl acetate lead citrate, and 70% showed this same characteristic rim-staining pattern after limited reaction (30 minutes) for peroxidase with diaminobenzidine. These distinctively large, rim-stained granules were identified in intact, mature peripheral blood neutrophils as well as in human bone marrow promyelocytes, indicating that their synthesis occurs during early myeloid development. This unusual granule type may play a specialized role in the microbicidal functions of the neutrophil, distinct from that of typical azurophil granules. PMID- 3040156 TI - Effect of warfarin on prothrombin synthesis and secretion in human Hep G2 cells. AB - Prothrombin synthesis and secretion were studied in a human hepatoma cell line (Hep G2) incubated with 35S-methionine for 2 to 24 hours at 37 degrees C. Extracellular and intracellular prothrombin were detected by immunoprecipitation with affinity-purified antiprothrombin antibody. Incorporation of 35S-methionine into prothrombin was monitored by counting specific bands excised from 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Prothrombin represented 0.3% to 0.7% of total newly synthesized protein secreted into the media. Warfarin had no effect on total prothrombin synthesis (extracellular plus intracellular). However, warfarin inhibited secretion of newly synthesized prothrombin by 58% to 73% over a 2 to 4 hour period. This was accompanied by the intracellular accumulation of an immunoprecipitable species of prothrombin of 78 kd, 6 kd less than extracellular prothrombin. At the end of the 4-hour incubation with warfarin, intracellular prothrombin increased from 44% to 82% (twofold) of total prothrombin, whereas extracellular prothrombin decreased from 56% to 19% (threefold) of total prothrombin. After 24-hour incubation with warfarin, intracellular and extracellular immunoprecipitable prothrombin approached control values. Deglycosylation of extracellular and intracellular prothrombin with hydrofluoric acid (HF) resulted in a decrease in mol wt for both species to 66 kd. Endoglycosidase-H treatment, which digests "early mannosyl" residues, resulted in a decrease in the mol wt of the intracellular species of 8 kd with no effect on the extracellular species. Thus, the lower mol wt intracellular species that accumulates following early warfarin treatment is due to the presence of incompletely processed carbohydrate chain. The data are compatible with the hypothesis that optimum glycosylation and secretion require Vitamin K-dependent carboxylation. PMID- 3040158 TI - Oestrogen receptor activity in intraduct and invasive breast carcinomas. AB - Breast cancers analysed for oestrogen receptor activity over a ten-year period have been surveyed in order to select a group of intraduct carcinomas without invasion and a second, control group of invasive carcinomas without intraduct carcinoma. Examination of histological sections taken from the face of the tumour samples used for receptor analysis showed only 13 purely intraduct carcinomas without any invasion. Each of these was matched for age, menstrual status, hospital of origin, and approximate assay date with two purely invasive ductal carcinomas of no specialized type (26 invasive carcinomas in all). In invasive carcinomas, a significantly higher proportion of the specimen was occupied by malignant cells (mean 30%) than in the intraduct carcinomas (mean 15%), and receptors were detected more frequently (77% versus 46%) and at higher concentrations (mean 26 times on a wet weight basis, 19 times on a protein basis). When allowance was made for the difference in cellularity between the groups, the invasive carcinomas still contained significantly higher concentrations of receptor protein (mean = ten times more on a wet weight basis). These findings suggest that the expression of the gene encoding the receptor protein tends to be a property either maintained, or acquired upon progression to invasive disease. Further studies will be needed to determine whether or not the established prognostic and predictive values of receptor measurements apply to non-invasive disease, and to clarify the relationship between receptor expression in benign and malignant breast in relation to morphological changes. PMID- 3040157 TI - Direct evidence for the interaction of the nucleotide affinity analog 5'-p fluorosulfonylbenzoyl adenosine with a platelet ADP receptor. AB - Previous reports have indicated that the nucleotide affinity analog 5'-p fluorosulfonylbenzoyl adenosine (FSBA) at concentrations between 40 and 100 mumol/L and at times greater than 20 minutes covalently modifies a single protein component on the external platelet membrane surface and that adenosine diphosphate (ADP) protects against this reaction. That this protein is an ADP receptor linked to platelet activation is shown by FSBA inhibition of ADP mediated platelet shape change, aggregation, and fibrinogen receptor exposure. In this report, further evidence for the interaction of FSBA with the ADP receptor on platelets is provided by the observation that FSBA at high concentrations (100 to 500 mumol/L) behaves as a weak agonist to produce platelet shape change within one minute as detected by spectroscopic assay and scanning electron microscopy with concomitant phosphorylation of the light chain of platelet myosin. The specificity of FSBA as an agonist is demonstrated by inhibition of FSBA-induced shape change by ATP and the covalent incorporation of SBA as well as the failure of 5'-fluorosulfonylbenozoyl guanosine (FSBG) to cause shape change. In contrast, incubation of platelets with low concentrations of [3H]-FSBA (40 mol/L) is not associated with stimulation of platelet shape change or myosin light chain phosphorylation. PMID- 3040159 TI - Cytochemical localization of thiamine pyrophosphatase and nicotinamide adenine dinucleotide phosphatase activities in rat epiphyseal chondrocytes. AB - Two phosphatase activities, which have been reported to be associated with the Golgi apparatus in several cellular types, have been cytochemically demonstrated in rat epiphyseal cartilage. This was the case for thiamine pyrophosphatase (TPPase) which was detected in Golgi trans face cisternae and also in nascent or immature secretory granules of chondrocytes. beta-Nicotinamide adenine dinucleotide phosphatase (beta-NADPase), on the other hand, was localized mainly in the endoplasmic reticulum region of both proliferative and hypertrophic chondrocytes. Most of the beta-NADPase reaction was shown to be associated with the cytoplasmic side of the rough endoplasmic reticulum membranes and also partially dispersed throughout the cytosol background. We suggest that beta NADPase in chondrocytes could be an enzyme with different properties from that described in other secretory cells. PMID- 3040160 TI - Interactions between normal lymphoid cells and HTLV-I producer cells. PMID- 3040161 TI - Extended use of the Mustarde dancing man procedure. AB - Webs other than epicanthal folds can be corrected by the Mustarde "dancing man" procedure. A series of 20 patients is presented in whom this method has been used to correct either minor degrees of congenital syndactyly or burn scar contractures in fingers. PMID- 3040162 TI - Restorative proctocolectomy: the four loop (W) reservoir. AB - The four loop (W) reservoir was developed with the aim of improving the functional results of the J reservoir and to avoid the need for catheterization often seen with the S reservoir. It is no more difficult to construct and results in better function. Sixty-four patients have undergone the procedure. There were no deaths. Mean hospital stay (including ileostomy closure) was 24 +/- 8 days. The reservoir has been removed in one patient (one week postoperatively) who was found to have a Dukes' B rectal carcinoma. Function has been assessed in 51 patients (mean follow-up 18.6 +/- 8.9 months). Frequency of defaecation per 24 h is 3.3 +/- 1.0 (range 1-8) with night evacuation in 14 per cent. Antidiarrhoeal medication is required by 20 per cent of patients. Continence is normal in 92 per cent, and 8 per cent have minor leakage. All patients defaecate spontaneously. PMID- 3040165 TI - Cytochrome reduction induced by increase in extracellular potassium and by electrical stimulation in isolated perifused neurohypophysis of guinea pig under mild hypoxic conditions. AB - Parallel reduction of cytochromes, a(a3), b and c + c1 was recorded under mild hypoxic conditions when the neurohypophysis was electrically stimulated and extracellular KCl concentration was increased. The electrically stimulated reduction was abolished by tetrodotoxin and by replacement of NaCl with LiCl. The reduction was not influenced by removal of extracellular Ca2+, or by application of a Ca ionophore, A23187. These results show that cytochrome reduction correlates with K+-influx in the resting state, and with Na+-influx followed by activation of the neurosecretory terminals contained in the neurohypophysis. PMID- 3040164 TI - Differential susceptibility to phosphatidylinositol-specific phospholipase C of acetylcholinesterase in excitable tissues of embryonic and adult Torpedo ocellata. AB - The ability of phosphatidylinositol-specific phospholipase C (PIPLC) to solubilize acetylcholinesterase (AChE) in the electromotor system of adult Torpedo ocellata and in the developing electric organ was examined. PIPLC solubilizes significant amounts of the membrane-bound G2 form of AChE throughout embryonic development of the electric organ, as it does in the adult electric organ, the AChE of which we have shown to contain covalently bound inositol in its membrane-anchoring domain. In the electromotor system of the mature fish, PIPLC solubilizes almost quantitatively the AChE dimer in the electromotor axon as in the electric organ itself, but the corresponding fraction in the electric lobe is almost totally resistant to the phospholipase. This finding implies that the covalently bound phosphatidylinositol is added concomitantly with axonal transport. A substantial part of the G2 form in back muscle is sensitive to PIPLC, whereas the G4 tetramer of Torpedo brain is completely resistant. PMID- 3040163 TI - The plasma kinetics of sulbactam-ampicillin administered to calves by the intramuscular and subcutaneous routes. PMID- 3040167 TI - GABAA receptor blockers reverse the inhibitory effect of GABA on brain-specific [35S]TBPS binding. AB - Thirteen substances previously reported to antagonize the electrophysiological effects of gamma-aminobutyric acid (GABA) on neurons also reversed the inhibitory effects of GABA on specific [35S]t-butylbicyclophosphorothionate ([35S]TBPS) binding to sites on rat brain membranes in vitro with a rank-order of potencies similar to those found in electrophysiological systems (R 5135 greater than pitrazepin greater than bicuculline greater than SR 95103 greater than securinine) confirming the earlier conclusion that GABA inhibits [35S]TBPS binding by acting allosterically on physiologically relevant GABAA receptors. Pitrazepin is the most potent of a series of mono N-aryl piperazines that block GABAA receptors. The new aryl amino pyridazine GABA derivative SR 95531 was about 3-fold more potent than bicuculline and 39-fold more potent than the structurally related SR 95103. Four known GABA antagonists have the same rank orders of potencies as convulsants and as reversers of GABA's inhibitory action on [35S]TBPS binding (bicuculline greater than securinine greater than theophylline greater than caffeine). Reversal of GABA-induced suppression of [35S]TBPS binding provides a simple method for further characterizing GABAA receptors linked to TBPS binding sites, and facilitates identification of convulsants and novel, perhaps selective, GABA antagonists. PMID- 3040166 TI - Regional distribution of mu, delta and kappa opioid receptors in human brains from controls and parkinsonian subjects. AB - The binding properties of mu and delta opioid receptors were investigated in several areas of human brain by using [3H]Tyr-D-Ala-Gly-(Me)Phe-Gly-ol and [3H]Tyr-D-Thr-Gly-Phe-Leu-Thr as respective selective ligands, while the totality of opioid receptors was measured by using [3H]etorphine as a non-selective agonist. Receptor densities were highest in cerebral cortex, amygdala and striatum, and lowest in the substantia nigra (pars compacta). In the different brain areas of patients with Parkinson's disease, the density and the proportion of the various opioid receptors were not significantly different from control subjects. PMID- 3040168 TI - Activity-induced elevation of cerebellar cyclic GMP occurs in the absence of climbing fiber pathways. AB - The inferior olivary nuclei (ION) of Sprague-Dawley rats were chemically lesioned with 3-acetylpyridine (3-AP), and the completeness verified by the lack of retrograde labeling of the ION following the injection of horseradish peroxidase (HRP) into the cerebellum. The effects of locomotor activity or of immobilization on cerebellar cyclic guanosine monophosphate (cGMP) levels were determined in control saline-treated and experimental, 3-AP-treated rats. Three subgroups of rats from both the control and the experimental groups of rats were required to swim for 60 s, immobilized for 60 s or unmanipulated before being killed by microwave irradiation, and the cerebella were collected for cGMP determination. There was no statistically significant difference in cGMP levels between immobilized and unmanipulated rats in either the experimental or control groups. Pretreatment with 3-AP reduced cerebellar cGMP levels in both the immobilized and the unmanipulated rats to 50% of those observed in the comparably treated groups of saline-treated controls. When compared to the corresponding group of unmanipulated rats, locomotor activity induced a significantly greater elevation of cerebellar cGMP in the experimental animals than in the controls (P less than 0.05). These results indicate that while inputs from the ION to the cerebellar Purkinje cells are probably important in maintaining the normal levels of cGMP seen in inactive rats, the locomotor-induced elevation of this parameter occurs in the absence of climbing fibers (from ION). The mechanisms responsible for the greater activity-induced elevation of cGMP levels seen in rats receiving 3-AP over control rats are discussed. PMID- 3040169 TI - Alpha 2-adrenergic receptors mediate inhibition of cyclic AMP production in neurons in primary culture. AB - The actions of adrenergic agents on the intracellular production of cyclic adenosine monophosphate (AMP) was examined in intact cortical and striatal neurons in primary culture, generated from the fetal mouse brain. Exposure of striatal neurons to the beta-adrenergic agonist isoproterenol (10 microM) resulted in a 5-fold increase in intraneuronal cyclic AMP; norepinephrine (100 microM), alone or in combination with isoproterenol, produced only a 3-fold increase in cyclic AMP levels. However, in the presence of yohimbine (10 microM), cyclic AMP productions due to norepinephrine or isoproterenol plus norepinephrine were identical to isoproterenol alone. When striatal or cortical neurons were exposed to pertussis toxin (100 ng/ml) overnight, there was no detectable difference between isoproterenol- and norepinephrine-stimulated cyclic AMP production. These data suggest that alpha 2-adrenergic receptors mediate the attenuation of cyclic AMP production in neurons and do so via the inhibitory guanine nucleotide regulatory protein of adenylate cyclase. PMID- 3040171 TI - Selective increase of alpha 1-adrenoceptors and muscarinic cholinergic receptors in rat cerebral cortex after chronic haloperidol. AB - The effect of chronic administration of haloperidol on alpha 1-, alpha 2-, and beta-adrenoceptors, cholinergic muscarinic, GABAA and benzodiazepine receptors in the cerebral cortex of the rat was investigated. Doses of 0.3 and 2 mg/kg of haloperidol during 7 days increased markedly the density of alpha 1-adrenoceptors without changes in affinity. The alpha 2- and beta-adrenoceptors were not modified after neuroleptic administration. The number of muscarinic receptors were also increased after haloperidol treatment (2 mg/kg/day). However, the GABAA and benzodiazepine binding sites remained unchanged. In the brainstem an increment in the alpha 1-, but not the beta-adrenoceptors was observed. The well known increase in the dopamine receptors in the striatum was confirmed. These observations demonstrate a multireceptor effect of haloperidol in the cerebral cortex. PMID- 3040170 TI - Rapid increase in brain benzodiazepine receptor binding following defeat stress in mice. AB - Defeat stress in mice, a model of social stress, increases benzodiazepine receptor binding as measured by specific [3H]Ro15-1788 binding in vivo, but not by [3H]flunitrazepam binding in vitro. This increase occurs rapidly, by 20 min following exposure to stress, and resolves by 60 min. Increased benzodiazepine receptor binding is observed in the cerebral cortex, cerebellum and hypothalamus, and appears to be due to an increase in receptor number rather than apparent affinity. The stress-induced increase in central benzodiazepine receptors is decreased in a dose-dependent fashion by lorazepam, a benzodiazepine agonist, but not by the receptor antagonist Ro15-1788. The stress-induced increase in benzodiazepine receptors is also blocked by adrenalectomy and is restored by corticosterone replacement. PMID- 3040172 TI - GABAergic synapses and benzodiazepine receptors are not identically distributed in the primate retina. AB - The distribution of benzodiazepine receptors (BZR) was compared to the distribution of gamma-aminobutyric acid (GABA)-ergic synapses in the rhesus monkey retina using monoclonal antibodies against the BZR and polyclonal antisera to glutamate decarboxylase (GAD), the GABA-synthesizing enzyme which labels the presynaptic terminals of the GABAergic synapses. Indirect immunofluorescence including dual fluorochroming for both BZR and GAD indicates that although both were localized to the inner plexiform layer and adjacent cell body layers, their distributions were largely non-overlapping. Thus, in the primate retina, BZRs are not exclusively associated with GABAergic synapses. PMID- 3040173 TI - A simplified method for monitoring the cytochrome aa3 redox state in bilateral cortical areas of unanesthetized cats. AB - We describe a versatile optical system that enables the simultaneous monitoring of the redox state of cytochrome c oxidase (cytochrome aa3) in two homologous cortical areas under chronically implanted windows in cats. A single light source, broad bandpass primary filters, light-conducting rods, and narrow bandpass interference detecting filters are employed. We observed reproducible responses of the cytochrome redox state and blood volume to carotid occlusion and terminal anoxia during anesthesia, and to graded doses of pentobarbital in awake animals. PMID- 3040174 TI - The effects of cholecystokinin and cholecystokinin antagonists on synaptic function in the CA1 region of the rat hippocampal slice. AB - The effects of two CCK antagonists, benzotript and proglumide, and of sulfated and non-sulfated cholecystokinin octapeptide (CCK-8-S and CCK-8-NS), were studied in the CA1 region of the rat hippocampal slice. Both benzotript and proglumide shifted presynaptic volley (prevolley) vs population spike input/output (I/O) curves for Schaffer collateral-commissural synaptic transmission to the right. This result indicates that the antagonists had a net depressant effect on synaptic transmission. CCK-8-S shifted prevolley vs population spike I/O curves to the left, indicating a net excitatory effect. Analysis of component I/O curves indicated that CCK-8-S and the CCK antagonists were acting postsynaptically by changing CA1 pyramidal cell threshold. CCK-8-NS had no significant effect on overall or component I/O functions. These findings suggest that endogenous CCK is released, directly or indirectly, upon stimulation of the Schaffer collateral commissural fibers and increases the excitability of CA1 pyramidal cells. PMID- 3040175 TI - Identification of a protease inhibitor produced by astrocytes that is structurally and functionally homologous to human protease nexin-I. AB - In the present studies we have compared the structural and biochemical properties of human protease nexin-I (PN-I) and a protease inhibitor present in the serum free culture fluid of normal rat brain astrocytes. The inhibitor binds to and forms covalent complexes with human urokinase and thrombin. The inhibitor has an approximate Mr = 43,000 based on the size of the complexes (deduced from SDS PAGE) and mediates the cellular binding and uptake of the proteases to which it links. Binding is heparin sensitive and occurs on a cell surface receptor that also binds complexes formed between proteases and a well-characterized cell secreted protease inhibitor, human PN-I. In addition, the inhibitor co-migrates with PN-I on SDS-PAGE and cross-reacts with anti-PN-I antibody on immunoblots. A similar molecule, designated NPF, is produced by C6 glioma cells in culture and has neurite promoting activity on a neuroblastoma cell line. PMID- 3040176 TI - Regional changes of brain Na+,K+-transporting adenosine triphosphatase related to ovarian function. AB - Synaptosomal Na+,K+-transporting ATPase activity of the mediobasal hypothalamus (MBH), the medial preoptic-suprachiasmatic (POSC) region and cerebral cortex was measured in rats at different stages of the estrous cycle and after ovariectomy and estradiol replacement. Enzyme activity of the MBH and POSC showed cyclic changes. In both regions it increased shortly before the proestrus surge of LH. However, the two areas responded to castration and estrogen treatment in an opposite fashion. No changes were detected in enzyme activity in the cerebral cortex. These findings are consistent with previous reports on cyclic changes in electrical activity and suggest that Na+,K+-ATPase activity could be a useful indicator of neural activity for the study of neuroendocrine interactions. PMID- 3040177 TI - Endogenous opioids regulate dendritic growth and spine formation in developing rat brain. AB - Continuous blockade of endogenous opioid-opioid receptor interaction by opioid antagonists from birth to day 10 increased neuronal maturation in the rat brain. The lengths of oblique dendrites of pyramidal cells in the cerebral cortex and basilar dendrites of the hippocampus were increased from controls by 136 and 51%, respectively, whereas the concentrations of spines in these cells were increased 183 and 69%, respectively. Total dendritic length of spiny branches of cerebellar Purkinje neurons was 65% greater than controls, and spine concentration of granule cells in the dentate gyrus was increased by 76%. Thus, endogenous opioids exert a remarkable influence on the timetable and magnitude of dendritic elaboration and spine formation, and serve as an important trophic influence in the regulation of neuro-ontogeny. PMID- 3040178 TI - Saphenous has weak ineffective synapses in sciatic territory of rat spinal cord: electrical stimulation of the saphenous or application of drugs reveal these somatotopically inappropriate synapses. AB - In a previous paper, we showed that chronic denervation of the sciatic nerve for more than 21 days in adult rats caused expansion of the saphenous nerve into sciatic territory in the spinal cord (medial L4, L5 and L6). To try to explain this expansion in the present paper, we tested the hypothesis that weak ineffective synapses of saphenous terminals are always present in sciatic territory. For this purpose the sciatic nerve was acutely denervated, the cord mapped with microelectrodes and responses evoked in single cells with natural (mechanical cutaneous) or electrical (pulses to saphenous nerve) stimulation. In the sciatic territory, no natural responses occurred but electrically evoked responses from the saphenous were everywhere. When drugs were applied to potentiate synaptic activity, many of the silent neurons in the sciatic territory in L4, L5 and L6 responded to natural inputs mediated by the saphenous. Picrotoxin was more effective than 4-aminopyridine which was more effective than strychnine in expressing these weak somatotopically inappropriate saphenous inputs. All together, these results support the hypothesis that weak ineffective saphenous inputs exist in sciatic territory of the spinal cord. They can be artificially expressed with electrical volleys or chemical potentiation and may be naturally expressed several weeks after chronic lesions of the sciatic nerve. PMID- 3040179 TI - Basal and electrically stimulated release of [3H]noradrenaline and [3H]dopamine from rat amygdala slices in vitro: effects of 4 beta-phorbol 12,13-dibutyrate, 4 alpha-phorbol 12,13-didecanoate and polymyxin B. AB - The protein kinase C activator 4 beta-phorbol 12,13-dibutyrate (PDB) enhanced in a concentration-dependent manner the electrically stimulated release of [3H]noradrenaline ([3H]NA) and [3H]dopamine ([3H]DA) from rat amygdala slices in vitro. PDB enhanced the basal release of [3H]NA and [3H]DA as well. 4 alpha Phorbol 12,13-didecanoate, which lacks the capacity to activate protein kinase C, was without effect on either basal or electrically stimulated release of [3H]NA and [3H]DA. Polymyxin B, which is a relatively selective protein kinase C inhibitor, decreased in a concentration-dependent manner the electrically stimulated release of both [3H]NA and [3H]DA from amygdala slices, whereas it enhanced the basal release of both neuromessengers. In the presence of 1.5 X 10( 7) M PDB, a concentration which when added to the superfusion medium alone doubled the electrically stimulated release of both [3H]NA and [3H]DA, polymyxin B again decreased in a concentration-dependent manner the release of both neuromessengers. At all polymyxin B concentrations used, the effect of the PKC inhibitor, expressed as percent inhibition, in the presence of PDB was approximately the same as that observed in the absence of PDB. This suggests that the antagonism between PDB and polymyxin B at the level of protein kinase C is not a competitive one. The effects of PDB and polymyxin B on basal release were additive. Taken together, these data suggest that in the amygdala presynaptically localized protein kinase C plays a role in signal transduction processes related to the exocytotic secretion of NA and DA from their nerve terminals. PMID- 3040180 TI - A model of chronic pain in the rat: high-resolution neuroanatomical approach identifies alterations in multiple opioid systems in the periaqueductal grey. AB - Inoculation of the tail base of rats with Mycobacterium butyricum led to an arthritic swelling and inflammation of the limbs which displayed a hyperalgesia to noxious pressure: these effects peaked at 3 weeks postinoculation. In vitro autoradiography of coronal sections of rat brain was used for a parallel determination of binding to mu-, delta- and kappa-opioid binding sites. In only two regions, the dorsomedial and dorsolateral parts of the periaqueductal grey (PAG), was a significant change seen: this comprised an increase in binding to kappa-sites, whereas mu- and delta-sites therein were unaffected. This region was analysed for opioid peptides derived from each of the three opioid peptide families known. While no change was seen in levels of immunoreactive (ir) dynorphin1-17 A (DYN) and ir-Met-enkephalin, a decrease was detected in those of ir-beta-endorphin (beta-EP): this change was restricted to the PAG. These data demonstrate a highly localized and selective influence of chronic arthritic pain upon multiple opioid systems in the PAG of the rat, a structure playing a key role in the control of pain and in the expression of the antinociceptive actions of opioids. The data suggest a possible significance of PAG pools of beta-EP and kappa-receptors in the response to and modulation of chronic pain. PMID- 3040181 TI - Evidence of lateral synaptic interactions in olfactory bulb output cell responses to odors. AB - Lateral inhibitory circuits are found throughout the nervous system. While the neuroanatomical basis for lateral inhibitory interactions exists in the olfactory bulb of Norway rats, there has been no direct demonstration of lateral inhibition in the responses of olfactory bulb output neurons to odor stimulation. In this report we recorded the extracellular activity of a large number of sequentially recorded mitral/tufted cells in response to odor stimuli at two different concentrations, as well as the inter-cell distance between these cells. The probability of recording two cells with excitatory responses to the same odor was then determined for inter-cell distances up to 500 microns. For cells stimulated with high concentration odors, the probability of two cells 100-200 microns apart both being excited by the same odor was significantly lower than that predicted if all cells responded independently. Cells separated by greater or shorter inter cell distances did not differ from the predicted value. Responses to the low odor concentration were not dependent on inter-cell distance. These results demonstrate that lateral synaptic interactions within the olfactory bulb influence output cell responses to odor stimulation. PMID- 3040182 TI - Endocrine cells in pancreatic islets of Langerhans are immunoreactive to antibody against guanine nucleotide-binding protein (Go) purified from rat brain. AB - Guanine nucleotide-binding proteins (G proteins) are usually classified into four subclasses (Gs, Gi, Go and Gt or transducin). We localized the anti-Go immunoreactivities in islets of Langerhans of the rat pancreas by using affinity purified antibody against the alpha-subunit of Go purified from rat brain. Endocrine cells of the islets of Langerhans were stained with this Go antibody, but the acinar cells in the exocrine portion of the pancreas were immunonegative to this antibody. These findings strongly suggest that Go protein functions as intermediaries in the transmembrane signalling pathway in the endocrine cells of the islets of Langerhans. PMID- 3040183 TI - Seizure-like discharges induced by lowering [Mg2+]o in the human epileptogenic neocortex maintained in vitro. AB - Seizure-like discharges were observed in slices of human epileptogenic neocortex maintained in vitro when [Mg2+]o was lowered near to zero. This type of epileptiform activity: (1) could occur spontaneously or following extracellular focal stimuli; (2) resembled the electrographic pattern associated with tonic clonic seizures; (3) was accompanied by increases in [K+]o (maximally 6.2 mM from a baseline of 3.25 mM) and decreases in [Ca2+]o (maximally 0.23 mM from a baseline of 1.8 mM). Application of the selective antagonist of N-methyl-D aspartate (NMDA) receptors, DL-2-amino-5-phosphonovalerate, suppressed in a reversible manner both spontaneous and stimulus-induced seizure-like discharges, suggesting that NMDA-activated conductances are important for the genesis of prolonged epileptiform discharges generated by human epileptogenic neocortical slices. PMID- 3040184 TI - Neuropeptide Y (NPY) in the area of the hypothalamic paraventricular nucleus activates the pituitary-adrenocortical axis in the rat. AB - Immunocytochemical studies have documented the presence of neuropeptide Y (NPY) in the hypothalamic paraventricular nucleus (PVN) which harbours a large number of neurones that contain corticotrophin-releasing factor (CRF). In this study the close morphological association between NPY fibres and CRF cell bodies in the PVN was confirmed. The localization of NPY terminals in the vicinity of CRF neurones forms a morphological basis for an action of NPY in the hypothalamic control of the pituitary-adrenocortical axis. We therefore microinjected NPY into the area of the PVN of both conscious, freely moving and anaesthetized rats and noted a powerful stimulatory effect on adrenocorticotropic hormone (ACTH) and corticosterone release as measured by radioimmunoassay. In experiments with conscious, freely moving rats, higher ACTH and corticosterone levels were detected following injection of NPY into the area of the PVN than following control injection (desamidated NPY). Intracerebroventricular injection of NPY produced a small, albeit significant, increase in circulating corticosterone levels as compared to control (saline-injected) rats. Anaesthetized rats responded to NPY (but not to saline) injected into the area of the PVN with elevated ACTH and corticosterone levels, while injection of NPY into the neocortex failed to affect the blood concentration of either ACTH or corticosterone. In conclusion, we have demonstrated an activating effect of NPY on the pituitary-adrenocortical axis both in conscious and anaesthetized rats which may reflect the anatomical relationship between NPY fibres and CRF neurones in the PVN. PMID- 3040185 TI - Sensitization occurs to the locomotor effects of morphine and the specific mu opioid receptor agonist, DAGO, administered repeatedly to the ventral tegmental area but not to the nucleus accumbens. AB - Several recent reports have demonstrated that opiate action in both the ventral tegmental area (VTA) and the nucleus accumbens (N.Acc.) produces an increase in locomotor activity. In the present experiments, the effect of repeated bilateral injections into these sites of either morphine or the mu opioid receptor agonist Tyr-D-Ala-Gly-NMe-Phe-Gly-ol (DAGO) was investigated. As previously reported with morphine and other opioids, repeated injections of either morphine or DAGO into the VTA produced a progressive enhancement or sensitization of their locomotor activating effects. On the other hand, although both substances injected into the N.Acc. elicited increased locomotion, repeated injections did not lead to sensitization. It has been suggested that the increased locomotor activity produced by opiate injection into the VTA is dopamine-dependent while that produced by intra-N.Acc. injections is not. The present findings provide neuroanatomical support for the view that sensitization to the locomotor activating effects of opiates and opioids brought about by repeated drug exposure involves the mesolimbic dopamine system. PMID- 3040186 TI - Evidence for a multiple innervation of cerebellar Purkinje cells by climbing fibers in adult ferrets infected at birth by a mink enteritis virus. AB - Newborn ferrets were inoculated with Mink Enteritis virus (parvovirus). They developed a cerebellar hypoplasia and presented severe ataxia. Electrophysiological study by intracellular recordings in the cerebellar cortex demonstrates that in these ferrets, like in other mammals, Purkinje cells deprived from granule cell input during development remain multiply innervated by climbing fibers in the adult. PMID- 3040187 TI - Hydrocortisone stimulates the development of oligodendrocytes in primary glial cultures and affects glucose metabolism and lipid synthesis in these cultures. AB - Cultures of glial cells were prepared from the brains of one-week-old rat pups. After one day in culture, serum was omitted from the medium and replaced by a combination of growth-stimulating hormones and other factors that enhanced the percentage of oligodendrocytes in the cultures. We investigated the effects of hydrocortisone on the development of oligodendrocytes, on the activities of oligodendrocyte-specific enzymes and on glucose- and lipid-metabolism of the glial cells. Hydrocortisone greatly enhanced the survival of glial cells in culture. The development of galactocerebroside-positive cells and the specific activity of 2',3'-cyclic-nucleotide 3'-phosphodiesterase were stimulated by 50 nM hydrocortisone, whereas these effects were partly reversed at higher concentrations of the hormone. The specific activity of glycerol-3-phosphate dehydrogenase was markedly stimulated by hydrocortisone; 1 microM or higher concentrations of hydrocortisone were required for an optimal effect. The consumption of glucose and the production of lactate were lowered by hydrocortisone whereas the oxidation of [6-14C]glucose to 14CO2 was not affected. Incorporation of [35S]sulfate into sulfolipids was greatly enhanced by hydrocortisone and [14C]incorporation from [1-14C]acetate into cholesterol and fatty acids was also stimulated but to a smaller extent. These results show that hydrocortisone exerts a general trophic function on glial cells in our culture system; enhances the ratio of oligodendrocytes over astrocytes, possibly by directing bipotential progenitor cells to develop into oligodendrocytes; specifically induces glycerol-3-phosphate dehydrogenase in oligodendrocytes. PMID- 3040188 TI - Cellular and synaptic physiology and epileptogenesis of developing rat neocortical neurons in vitro. AB - The cellular and synaptic physiology of developing rat neocortical neurons was studied using the in vitro slice method. Rats aged 1-28 days were used for analysis. During the first two postnatal weeks several sequential changes occur in membrane properties and evoked synaptic potentials. Immature neurons had higher input resistances, more linear I-V characteristics, longer membrane time constants, and slower rising and falling phases of action potentials. The developmental increase in rate of rise of the action potential suggests an increasing density of voltage-dependent Na+-channels are inserted in neuronal membranes during postnatal development. The higher input resistance of young cells might be due to their small size and differences in membrane properties. The long time constant indicates a higher specific membrane resistivity of immature neurons. Postsynaptic potentials (PSPs) recorded in young neurons were longer in latency, longer in duration, and more fragile during repetitive activation than their mature counterparts. In addition, PSPs evoked in neurons of animals less than 1 week old did not contain inhibitory postsynaptic components. These physiological features of immature neocortical neurons help explain the pattern of epileptogenesis in young animals. When neonatal cortical slices were exposed to the gamma-aminobutyric acid (GABA) antagonists penicillin or bicuculline, the frequency of occurrence of discharges resembling epileptiform depolarization shifts approached that found in mature slices only during the second postnatal week. Depolarization shifts at younger ages were less stereotyped and more sensitive to stimulus parameters than those in mature neurons. PMID- 3040189 TI - Loss of polyneuronal innervation and establishment of a topographical map in the glutaeus muscle of Bufo marinus during generation of secondary muscle cells. AB - The development of synaptic connections to the toad (Bufo marinus) glutaeus magnus from segmental nerves 8 (N8) and 9 (N9) was determined in the postmetamorphic period. Three different-size toads were studied: small (0.3-2.0 g), medium-size (5-15 g) and large (greater than 20 g). The number of cells in the glutaeus increased about 9-fold during development; this involved the appearance and subsequent maturation of secondary fibres throughout the muscle. The glutaeus in small toads, which consisted almost entirely of primary fibres, was innervated to a similar extent by N8 and N9 as assessed by tetanic contraction measurements. During late development there was a progressive increase in the percentage of the muscle innervated by N9 and a decrease in the percentage innervated by N8. This change in the segmental innervation was accompanied by changes in the innervation of the ventral glutaeus as assessed by intracellular recording. In small toads this surface of the muscle was innervated predominantly by N8, with N9 frequently appearing as a low-efficacy terminal on dually innervated fibres. With further development there was a progressive reduction in the percentage of dually innervated fibres and a concomitant decrease in the percentage innervation of the entire ventral glutaeus by N8. These results suggest that the topographical projection is established by the initial distribution of N9 terminals on the primary fibres of the muscle. The multiple innervation of newly generated fibres and the on-going process of terminal elimination results in N9 terminals, many of which were initially weak, preserving their position in the muscle. This occurs at the expense of N8 terminals, whose relative incidence declines during development. The competitive advantage of N9 motoneurones may be due to their greater capacity to lay down axon collaterals and preferentially innervate newly generated fibres; alternatively N9 terminals may displace N8 terminals, which were initially more efficacious, from dually innervated fibres. Secondary muscle fibres generated throughout the muscle are thus incorporated into an increasingly precise topographical map. PMID- 3040190 TI - Development of cytochrome oxidase staining in the retina and lateral geniculate nucleus: a possible correlate of ON- and OFF-center channel maturation. AB - A histochemical stain for cytochrome oxidase (CO) activity was used to examine the maturation of a neurochemical correlate of ON and OFF channels in the retina and dorsal lateral geniculate nucleus (LGN) of the tree shrew. In the adult tree shrew, the CO staining pattern can be used as a histochemical marker of segregated ON- and OFF-center channels in the retina, LGN, and striate cortex. Our previous studies have shown that the retina is immature and the LGN unlaminated at birth. In the present study, we show that the laminar development of CO reactivity emerges during the first postnatal week in the LGN, while the maturation of CO staining in the presumed ON and OFF sublaminae of the retinal inner plexiform layer develops slowly, well after the appearance of differential laminar CO staining in the LGN. PMID- 3040191 TI - Development of neuronal activity in the rat suprachiasmatic nucleus. AB - The development of function was studied in the rat suprachiasmatic nucleus (SCN) by analyzing the rate and pattern of neuronal discharge in vitro from embryonic day 22 (E22) to postnatal day 14 (P14), and comparing these with adult SCN. The firing rate of SCN neurons on E22 is low but there is a clear circadian rhythm with subjective day values (1.6 +/- 0.2 impulses/s) significantly higher than those for subjective night (0.9 +/- 0.2 impulses/s). At E22 most neurons show an irregular firing pattern. The firing rate of SCN neurons gradually increases from E22 to P14 with emergence of patterns of firing of individual neurons that approximate those of adult SCN. In calcium-free medium the firing rate of SCN neurons at P1 is reduced and the circadian rhythm in firing rate is abolished. These results demonstrate that SCN neurons exhibit a circadian rhythm in firing rate early in development, largely prior to the formation of synaptic contacts within the SCN, and indicate that expression of the rhythm requires the presence of calcium ions. PMID- 3040193 TI - Brain spectrin, calpain and long-term changes in synaptic efficacy. AB - This chapter discusses the possibility that proteolytic digestion of cytoskeletal proteins, in particular spectrin, is part of the mechanisms through which physiological activity elicits structural and chemical changes in brain synapses. Recent work from several laboratories has produced a description of the initial events that trigger the long-term potentiation (LTP) of synaptic responses that appears in hippocampus after brief episodes of high frequency electrical stimulation. A likely sequence is as follows: suppression of IPSPs, prolongation of EPSPs, activation of N-methyl-D-aspartate (NMDA) receptors, influx of calcium into target cells. After briefly describing the evidence for this triggering sequence, the review takes up the question of what types of calcium sensitive chemistries are available to synaptic region that could produce functional changes lasting for weeks (i.e., for LTP). It is argued that the partial degradation of spectrin by a calcium-activated protease (calpain) provides a mechanism of this type. Spectrin is a substrate for calpain and both it and a breakdown product comparable to that produced by calpain are found in postsynaptic densities. Moreover, there is substantial evidence that spectrin regulates the surface chemistry and morphology of cells and thus its partial degradation would be expected to produce pronounced and persistent modifications in synapses. To reinforce this point, the review discusses recent findings suggesting that calpain mediated proteolysis of spectrin and other cytoskeletal proteins produces substantial changes in the shape of blood-borne cells and the distribution of their surface receptors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3040192 TI - Influence of rostral and caudal brain stem reticular formation on thalamic neurons. AB - Single neuronal activity was recorded from the diffuse thalamic system. Influence of the rostral desynchronizing and caudal synchronizing structures of the brain stem reticular formation on these neurons was studied. Rostral stimulation produced an increase and caudal stimulation a decrease in the thalamic unit firing. A possible mechanism by which the brain stem reticular structures influence the cortical neurons is proposed on the basis of these findings. PMID- 3040194 TI - [Murine herpes viruses, their experimental pathogenesis and ecologic significance]. PMID- 3040195 TI - [Multicenter nutritional survey of possible etiologic factors in chronic tropical pancreatitis]. PMID- 3040196 TI - [Natural substances regulating fertility. Effect of plant extracts in the Ivory Coast pharmacopoeia on the release of LH by hypophyseal cells in culture]. AB - The in vitro action of hydro-alcoholic extracts of plants from Ivory Coast pharmacopoeia was analyzed on cultured rat pituitary cells. Cells were treated for 24 hours with various doses of extracts and then stimulated for 4 hours with 10(-8) M LHRH. Extracts from Afrormosia laxiflora, Cola nitida, Pterocarpus erinaceus and Tetrapleura tetraptera inhibit the LHRH-induced release of LH. On the contrary, extract from Combretodendron africanum stimulates the basal release of LH and this increase is added to the LHRH-induced release. Therefore, the natural substances contained in these plants may in vitro exert a regulation of the gonadotropin release. PMID- 3040197 TI - Equine herpes myeloencephalopathy. AB - The neurologic form of EHV-1 infection appears to be the result of central nervous system infarction caused by vasculitis, which is initiated in endothelial cells of small blood vessels. The etiologic agent is equine herpesvirus-1, subtype 1. There is some evidence to suggest that the neurologic form of the disease actually results from reactivation of a previous infection. Whether the vasculitis that causes the central nervous system injury is the direct result of the infection or an immune response to the infection has not been determined. The clinical signs are rapid in onset, nonprogressive, and many horses may improve. The diagnosis must often remain tentative, particularly in horses that recover, because there is no single reliable confirmatory test. The prognosis is generally good, although recovery may be slow and incomplete. Supportive therapy is essential, and administration of corticosteroids may be useful. There is no specific therapy for the virus or for the vasculitis. Currently no vaccine can be claimed to protect against the central nervous system form of the disease. Vaccination is recommended, however, to reduce the incidence of respiratory disease, abortion, and neonatal death on the farm. Repeated vaccination is necessary to maintain presumably protective antibody concentrations. Vaccination every 3 to 4 months may decrease the incidence of EHV-1 infection on the farm and therefore may indirectly prevent the occurrence of the neurologic form of the disease. PMID- 3040198 TI - Neuritis of the cauda equina. AB - Although the specific etiology of NCE remains unknown, many advances have been made in recent years. It is hoped that we can expect continued success in this area. Etiologic and pathophysiologic determination of NCE is the key to developing an appropriate therapeutic regimen. PMID- 3040200 TI - Effects of aluminum(III), chromium(III), and iron(III) on the rate of dissolution of calcium hydroxyapatite crystals in the absence and presence of the chelating agent desferrioxamine. AB - Aluminum ions (Al) and chromium (III) ions (Cr), as they exist in aqueous solution at neutral pH, adsorb onto calcium hydroxyapatite crystals (HAP) and severely inhibit their dissolution process, when present in concentrations less than 1 microM. Iron (III) ions (Fe), at concentrations up to 10 microM, have no effect on the dissolution process of HAP. The Fe-chelating agent desferrioxamine also forms strong complexes with Al but not with Cr. Desferrioxamine prevents the adsorption of Al to HAP and removes pre-adsorbed Al from the HAP surface, although not instantaneously, but has no significant effect on the adsorption of Cr to HAP. Desferrioxamine is also found to be capable of removing Al preadsorbed to bone powder. PMID- 3040199 TI - Calcium phosphate phase transformations in serum. AB - A better knowledge of the pathological calcification mechanisms should provide a rational basis for their control. In the present study, dicalcium phosphate dihydrate (DCPD, CaHPO4 X 2H2O) was used as a source of calcium and phosphate ions to investigate the mechanism of formation of more basic and more insoluble calcium phosphates in ultrafiltered serum (u.f.s.). DCPD crystals were suspended in u.f.s. at 37 degrees C by constant stirring; samples were removed periodically for calcium and phosphate analysis and pH measurement. Occasionally, samples of solids were removed for X-ray diffraction. The experiments were carried out both with and without a 5.5% CO2 atmosphere. After initially becoming saturated with DCPD, the u.f.s. composition changed and became saturated with respect to octacalcium phosphate (OCP, Ca8H2(PO4)6 X 5H2O). At this point OCP crystals were detected in the solid phase by X-ray diffraction. Further stirring changed the composition so that it became undersaturated with both DCPD and OCP and shifted toward, but did not reach, a value so low as to be saturated with hydroxyapatite (OHAp, (Ca5(PO4)3OH). The presence of CO2 in the atmosphere slowed down, but did not prevent, the above sequence of events. The above results strongly suggest that calcifications, beneficial and pathological, that take place in serum may involve OCP as a precursor, which hydrolyzes in situ to a more basic apatitic product. The results also indicate that direct formation of OHAp in u.f.s. is a very slow process and may occur only rarely. The process appears to be similar in whole serum. PMID- 3040201 TI - Effects of vitamin D3, 25-hydroxyvitamin D3, and 24,25-dihydroxyvitamin D3 on parathyroid hormone secretion. AB - The active vitamin D metabolite 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] causes marked suppression of both pre-proparathyroid hormone messenger RNA (pre-proPTH mRNA) and parathyroid hormone (PTH) secretion. These effects are dose dependent and reversible when tested in an in vitro primary tissue culture cell system using normal bovine parathyroid cells. In the current studies, the precursors of 1,25(OH)2D3 and the related metabolite 24,25-dihydroxyvitamin D3 [24,25(OH)2D3], were used in the same culture system to test for possible regulatory effects. The results were compared with identically prepared cells exposed to 1,25(OH)2D3. In short-term studies (30-120 minutes), none of the vitamin D-related compounds produced any effect on PTH secretion. In long-term studies (24-48 hours, using primary tissue culture in the presence of test agents), neither vitamin D3 nor 25(OH)D3 affected PTH secretion or pre-proPTH mRNA over the concentration range 10(-11)-10(-7) M. On the other hand, 24,25(OH)2D3 produced significant suppression of both pre-proPTH mRNA (77% of control, P less than .01) and PTH secretion (75% of control, P less than .005) at 10(-7) M. By comparison, 10(-11) M 1,25(OH)2D3 produced levels of suppression (25-30%) of both pre-proPTH mRNA and PTH secretion comparable to 10(-7) M 24,25(OH)2D3, while even greater suppression (40-50%) occurred at 10(-9)-10(-7) M 1,25(OH)2D3. From these studies, we conclude that vitamin D3 and 25(OH)D3 do not have significant effects on PTH synthesis and secretion over the range of doses tested.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3040202 TI - Ionic changes induced by excitatory amino acids in the rat cerebral cortex. AB - The ionic mechanisms underlying the action of excitatory amino acids were investigated in the rat motor cortex. Ion-selective microelectrodes were attached to micropipettes such that their tips were very close and local changes in extracellular concentration of sodium, calcium, and potassium ions elicited through ionophoretic applications of glutamate (Glu) and of its agonists N-methyl D-aspartate (NMDA), quisqualate (Quis), and kainate (Ka) were measured. These agents produced moderate increases in [K+]o (up to 13 mM) but, in contrast, substantial tetrodotoxin-insensitive decreases in [Na+]o (maximally of 60 mM). NMDA-induced sodium responses could be blocked by manganese, while the Quis- and Ka-induced responses were not. Quis and Ka produced increases in [Ca2+]o or biphasic responses while NMDA, even with small doses, induced each time drastic decreases in [Ca2+]o (maximally of 1.15 mM), which could be attenuated or blocked by manganese but not by organic calcium channel blockers. NMDA responses could be abolished by reduced doses of 2-amino-phosphonovalerate. The largest Glu- and NMDA-induced calcium responses were observed in the superficial cortical layers, but such maxima disappeared after selective degeneration of pyramidal tract neurons. All amino acids produced sizeable reductions in the extracellular space volume. The following can be concluded. (i) All the excitatory amino acids tested induce an increased permeability to sodium and potassium ions. (ii) In addition, the NMDA-operated channels have specifically a large permeability for calcium, although calcium ions contribute only by less than 10% to the NMDA-induced inward currents.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3040203 TI - Dexamethasone inhibits ovine corticotrophin-releasing factor (oCRF), arginine vasopressin (AVP), and oCRF + AVP stimulated release of ACTH during the last third of pregnancy in the sheep fetus. AB - We examined the hypothesis that in fetal sheep during late pregnancy exogenous glucocorticoids might affect differentially the pituitary response, measured as changes in plasma ACTH concentrations, to the systemic administration of ovine corticotrophin-releasing factor (oCRF), arginine vasopressin (AVP), or oCRF + AVP. At d 113-116 of pregnancy, equimolar injections of oCRF and AVP given separately provoked similar significant increases in plasma ACTH; the change in ACTH over basal values was significantly greater than the sum of the two separate responses when AVP + oCRF were given together. Exogenous dexamethasone did not affect basal ACTH concentrations, but suppressed significantly the responses to oCRF, AVP, and oCRF + AVP. At d 126-130, there was a significant ACTH response to CRF alone and to AVP + oCRF, but not to AVP alone. The response during the first 30 min postinjection to oCRF was significantly less than that to AVP + oCRF. Plasma cortisol rose after each peptide injection. Exogenous dexamethasone suppressed both basal and stimulated responses to each peptide. At the amounts injected, there was no significant ACTH or cortisol response to oCRF, AVP, or oCRF + AVP at d 136-140, but dexamethasone suppressed basal ACTH and cortisol concentrations at this time. We conclude that stimulated, but not basal, release of ACTH is subject to the negative feedback effect of exogenous glucocorticoid by d 113-116 of gestation in fetal sheep. Both basal and stimulated release of ACTH and cortisol are suppressed after d 125. At the amount of exogenous dexamethasone given, oCRF, AVP, and oCRF + AVP-stimulated responses are affected similarly.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3040204 TI - Detection and quantitation of human enteric viruses in waste waters: increased sensitivity using a human immune serum globulin--immunoperoxidase assay on MA-104 cells. AB - This study demonstrates that the most sensitive method for the detection and quantitation of cultivable human enteric viruses in water samples after repassage in the MA-104 cell line is the detection of infected cells by the human immune serum globulin--immunoperoxidase (HISG-IP) method recently described by the authors. This immunoperoxidase method is up to 50 times more sensitive than a liquid overlay assay by cytopathic effect in BGM cells. The viral content of waste waters was evaluated with this new methodology. By this method the average viral content of raw sewage (RS) was 900 mpniu/L (most probable number of infectious units per litre), 1056 mpniu/L in primary effluent (PE), and 106 mpniu/L in secondary effluent (SE). With a cytopathic effect assay on BGM cells, values of 85 (RS), 56 (PE), and 2 (SE) mpniu/L were observed, a striking underestimation of the viral content of secondary effluents. PMID- 3040205 TI - Second-step reconcentration of environmental samples by ammonium sulfate flocculation of beef extract. AB - Some enteric viruses are sensitive to the acid environment utilized during the concentration of viruses from water. The use of a neutral flocculant, neutralized ammonium sulfate at 50% saturation, permitted the recovery of 97% of the simian rotavirus SA-11, 87% of Coxsackievirus B-4, and 88% of poliovirus type 1. This method should permit a better recovery of enteric viruses from the environment. PMID- 3040206 TI - Association of germ cell tumours of the testis and intrathoracic sarcoid-like lesions. PMID- 3040207 TI - 5-Fluorouracil, doxorubicin, and mitomycin-C in the treatment of adenocarcinoma and large cell carcinoma of the lung. AB - Fifty-four patients with adenocarcinoma and large cell carcinoma of the lung, 16 with localized surgically resected disease and 38 with metastatic disease, were treated with 5-fluorouracil, doxorubicin, and mitomycin-C (FAM), in an attempt to prolong previously reported survival times in the patients. The postsurgical patients were given elective chemotherapy. Eighteen received radiation therapy for control of local disease. Tumor regression, attributable to chemotherapy alone, was noted in ten of 20 cases in whom there was measurable disease. The median survival time of all patients was 32 weeks, ranging from 25 weeks in those with distant metastases to 56 weeks in patients with surgically resected disease. FAM did not seem to produce any beneficial survival advantages. PMID- 3040208 TI - Relapse-associated variables in stage I favorable histology Wilms' tumor. A report of the National Wilms' Tumor Study. AB - All 24 cases of confirmed and evaluable Stage I favorable histology (FH) Wilms' tumor (WT) relapsing to date on the Third National Wilms' Tumor Study (NWTS-3) were compared with 48 matched control subjects who had not relapsed for at least 2 years after diagnosis. Fifteen of the clinical and pathological variables studied, including patient age and tumor size, had no significant relationship to the outcome of this study. Four histologic features, all related to the degree of tumor extension within the "tumor-kidney unit" proved to be significantly associated with relapse. These were (1) invasion of the tumor capsule, (2) presence of an "inflammatory pseudocapsule," (3) renal sinus invasion, and (4) tumor in intrarenal vessels. One or more of these features was present in 100% of relapsed cases (excluding one for which two variables were unevaluable), but occurred in only 46% of controls (P less than 0.0002). Therefore, no relapses occurred in the NWTS-3 when all four variables were negative. This result was confirmed by a review of all Stage I cases in the NWTS-1 who had relapsed and who were treated by single-agent chemotherapy. Again, no relapses occurred when all four factors were negative. These results demonstrate the feasibility of "microsubstaging" and could serve as a basis for future refinements of therapy for Stage I favorable histology Wilms' tumor. PMID- 3040209 TI - Radiotherapy in the management of pancreatic islet cell tumors. AB - Malignant islet cell tumors are commonly treated with surgical resection. Chemotherapy is reserved for residual, unresectable, or metastatic disease. The role for radiotherapy has not been clearly defined. This article describes three cases of advanced islet cell tumors treated effectively with radiotherapy. This experience, in addition to that from other published reports, suggests that radiotherapy is a useful mode for treating advanced islet cell carcinoma. PMID- 3040210 TI - Radiation-induced long thoracic nerve palsy. AB - The incidence of long thoracic nerve palsy after radical mastectomy has been documented to be approximately 10%. No cases have been reported after the more recent treatment for breast cancer, lumpectomy with axillary dissection. This more recent surgical procedure is customarily followed by aggressive radiation therapy to the remaining breast tissue. This is the first case report of a patient with radiation-induced long thoracic nerve palsy. The patient was a young woman who underwent left breast quadrantectomy and axillary dissection for breast cancer. After radiation therapy, she had isolated left long thoracic nerve palsy. The diagnosis was confirmed by electrodiagnostic studies. Almost full recovery occurred after 5 months. PMID- 3040211 TI - Type IV collagenase activity of a primary HSV-2-induced hamster fibrosarcoma and its in vivo metastases and in vitro clones. AB - The expression of a basement membrane (BM) collagen-degrading metalloprotease (Type IV collagenase) was studied in a herpes simplex virus (HSV)-2 transformed hamster fibrosarcoma and its in vivo derived sublines and in vitro derived clones of varying metastatic potential. The primary parent tumor was shown to release more or less Type IV collagenolytic activity compared with its sublines (derived from lung nodules that developed after resection of the primary tumor). Normal baby hamster kidney and hamster embryo fibroblasts did not secrete detectable amounts of BM collagenase, whereas normal hamster lung fibroblast secreted intermediate levels of Type IV collagenase activity. The collagenase IV activity of the parent tumor and its in vivo and in vitro derived sublines was assayed in vitro and compared with the ability of the cells lines to spontaneously metastasize in vivo. No correlation between the ability to secrete type IV collagenase and metastatic propensity was detected. Although all cell lines secreted type IV collagenase, the highest activity was recorded for a nonmetastatic variant. PMID- 3040213 TI - Histologic malignancy grading of invasive ductal breast carcinoma. A regression analysis of prognostic factors in low-risk carcinomas from a multicenter trial. AB - In a prospective adjuvant trial including patients with primary operable breast cancer, invasive carcinomas of ductal type were subjected to histological malignancy grading. The parameters investigated were tubule formation, number of mitoses and cell pleomorphism. A Cox regression model for survival data was applied to evaluate the impact of the histological parameters on prognosis in 1809 patients with low-risk carcinomas. Cell pleomorphism proved superior to the other histologic tumor characteristics. It was found that low-risk invasive ductal carcinomas with severe cell pleomorphism had an excess recurrence intensity of 209% relative to carcinomas with no pleomorphism. It is therefore suggested that polymorphous invasive ductal breast carcinomas, other things being equal, should be regarded as high-risk tumors in future clinical trials. Finally it was found that the tripartite malignancy grading 1, 2 and 3 characterizing each of the histological parameters was not equidistant. Consequently, the traditional tripartite histologic scoring needs reconsideration. PMID- 3040212 TI - Tumor-associated antigens in breast carcinomas. Prognostic significance. AB - In an attempt to identify biologic markers that might predict prognosis in breast cancer patients, the presence or absence of seven tumor-associated antigens in 54 infiltrating breast carcinomas was correlated with tumor recurrence rates (minimum five-year follow-up), axillary lymph node metastases and tumor volume. Immunohistochemical kappa-casein was present in 30 (56%) tumors, alpha lactalbumin in 39 (72%) tumors, secretory component of IgA in 26 (48%) tumors, carcinoembryonic antigen in 34 (63%) tumors, pregnancy-specific beta-1 glycoprotein in 7 (13%) tumors, beta subunit of human chorionic gonadotrophin in 1 (2%) tumor and human placental lactogen in 0 (0%) tumors. There was no significant correlation between the presence or absence in tumor of any of the antigens, and prognosis as assessed either by 5-year recurrence rates (P greater than 0.18) or by the presence of axillary lymph node metastases (P greater than 0.20). No significant difference was noted in mean tumor volume (cm3) +/- SEM, between tumors with or without antigen immunoreactivity (P greater than 0.05). PMID- 3040214 TI - Lung carcinoma in the elderly population. Influence of histology on the inverse relationship of stage to age. AB - Unlike most cancers, lung carcinoma is more likely to be localized at the time of diagnosis in older age groups when compared to those who are middle-aged. In an attempt to explain this inverse age-stage relationship we studied 9062 histologically confirmed cases of lung carcinoma occurring from 1975 to 1984 obtained from the regional cancer registry for Kansas and western Missouri. They were analyzed according to histologic type, age, sex, and stage at diagnosis. The data suggest that the proportion of squamous cell carcinoma rises and that of adenocarcinoma and small cell undifferentiated carcinoma falls with increasing age. The proportion of staged lung carcinoma with local disease at the time of diagnosis increases with age. In males this trend occurred in all cell types except large cell undifferentiated carcinoma but was most significant for squamous cell carcinoma. Squamous cell carcinoma was the only group to show a significant trend among females. The rise in squamous cell and fall in small cell carcinoma may partially explain the increased prevalence of local stage disease with advancing age. PMID- 3040215 TI - Mucoepidermoid tumors of the lung. AB - Mucoepidermoid tumors of lung (MET) are rare tumors derived from the minor salivary gland tissue of the proximal tracheobronchial tree. The authors studied 58 cases of MET confined to the lung and used criteria derived from similar tumors of the salivary glands to separate them into low-grade and high-grade variants. The overwhelming majority of low-grade tumors behaved in a benign fashion, whereas 23% of high-grade tumors resulted in patient death. Prognostic factors which appeared to predict future aggressive behavior included high-grade classification, advanced stage at presentation, and perhaps lymph node metastases. PMID- 3040217 TI - Fulminant leukemic polyradiculoneuropathy in a case of B-cell prolymphocytic leukemia. A clinicopathologic report. AB - A 52 year old man with a 10-month history of B-cell prolymphocytic leukemia (PLL) died of an apparent acute fulminant polyradiculoneuropathy, a condition generally attributed to paraneoplastic complication. The pathologic examination disclosed diffuse leukemic infiltrations of the peripheral nervous system. It is suggested that this particularly aggressive form of B-cell chronic prolymphocytic leukemia presented a constellation of features that promoted the invasion of the peripheral nervous system by way of the bloodstream and may explain the unusual clinical presentation. PMID- 3040216 TI - Primary liver cancer in Japan. Sixth report. The Liver Cancer Study Group of Japan. AB - The Liver Cancer Study Group of Japan statistically analyzed 4658 cases of primary liver cancer diagnosed from January 1, 1980 to December 31, 1981 in over 400 hospitals throughout the country. The study group comprised 2038 cases of hepatocellular carcinoma, 146 of cholangiocarcinoma, 33 of mixed carcinoma, 30 of hepatoblastoma, six of sarcoma, and 33 others. In 2286 cases (49.1%) a histologic diagnosis was available. The survey, based mostly on the histologically proven cases, describes histologic features of the tumors, grade of anaplasia and growth patterns of the tumor cells, pathology in noncancerous portions of the liver, distant metastases, medical history, frequency of hepatitis in the history, frequency of positive HBsAg and anti-HBs, age distribution, subjective symptoms, radiographic features (angiogram, scintiscan, computed tomography), ultrasonography, surgical procedures, extent of hepatic resection, and survival. PMID- 3040218 TI - Human immunodeficiency virus-associated T-cell lymphoblastic lymphoma in AIDS. AB - A patient with multiple infections whose serum had antibodies to a human immunodeficiency virus (HIV) developed a T-cell lymphoblastic lymphoma (T11+, Leu 1+, Leu-3a+, TdT+, B1-, common ALL-). Antibodies to human T-lymphotropic virus-I (HTLV-I) were absent. T-cell leukemia-lymphoma may be associated with HIV infection. PMID- 3040219 TI - The histologic response of soft tissue sarcoma to radiation therapy. AB - Twenty-seven patients with soft tissue sarcoma had preoperative radiotherapy, limb-sparing marginal surgical resection and whole-mount tumor histologic analysis. Incisional biopsy specimens before radiotherapy were reviewed for tumor type, grade, and extent of necrosis. Preoperative radiotherapy was given in either of two regimens: 13 patients received a mean total dose of 5250 cGy in one daily 180 to 200 cGy fractions and 14 patients a mean total dose of 4770 cGy in two daily fractions of 180 to 200 cGy separated by 4 hours. Twenty-one specimens had at least 80% necrosis or severely altered cells, a 3+ to 4+ response. Grade and size of the tumor appeared to be indicators of response to treatment rather than histologic type. Three of five patients (60%) with Grade 1, eight of 11 patients (73%) with Grade 2 lesions, and ten of 11 patients (91%) with Grade 3 tumors had 80% or greater necrosis or severely altered cells. For tumors 10 cm or less in greatest diameter, the 3+ to 4+ histologic response was seen in 12 of 14 patients (86%) whereas for lesions greater than 10 cm, this response was observed in nine of 13 patients (69%). For patients with Grade 2 or 3 soft tissue sarcoma, 13 of 14 patients (93%) treated with two fractions per day and two of four patients (50%) receiving one fraction per day exhibited significant response. All six patients treated twice daily for lesions greater than 10 cm had 3+ to 4+ histologic response compared to three of seven (43%) patients treated once per day. Therefore, grade and size of soft tissue sarcoma are important predictors of response to radiotherapy and preoperative twice daily radiotherapy may more likely permit the conservative surgical excision of sarcomas of borderline resectability. PMID- 3040220 TI - EPR spectroscopic analysis of binding sites of a cancerostatic agent on erythrocyte membranes. AB - A biologically highly active antitumor agent, chloroethyl-nitrosourea-piperidine N-oxyl, was found to bind covalently to sulfhydryl groups of membrane proteins of erythrocytes. The agent penetrates the erythrocyte membrane very easily within three minutes. Mainly two different binding sites are distinguishable with rather high rotational mobility of the NO-moiety and high polarity of their environment. The lowered broadening of bound label suggests pocket-like binding sites with a limited accessibility for paramagnetic ions. Besides common concepts that antitumor agents often bind to DNA and inhibit cell proliferation, further targets on plasma membranes are taken into consideration for antitumor agents. PMID- 3040221 TI - Characteristic patterns of chromosome abnormalities in acute myeloid leukemia with Auer rods. AB - Two hundred and four cytogenetically investigated patients with acute myeloid leukemia (AML) were evaluated for the presence (AR+) or absence (AR-) of Auer rods. Chromosome analysis was successful in 187 patients (92%). Seventy-eight patients (38%) were AR+. Cytogenetic abnormalities were detected in 35 (49%) AR+ patients and in 66 (57%) AR- patients. The proportion of patients with complex karyotypic changes (more than two aberrations) was significantly higher in the AR group (p less than 0.01). Also, unidentifiable marker chromosomes were significantly more frequent in the AR- group (p less than 0.01). Twenty-one of 23 AR+, but none of 46 AR- patients with structural changes, had involvement of 21q22, 17q11-12, or 11p13-15. In contrast, structural changes of 1p, 5q, 7q, 9q, 11q, or 22q were present in 31 AR- but in only two AR+ patients. Four patients had translocations involving 21q22 without rearrangements of 8q22. All were AR+, but only one displayed M2 morphology. We draw the conclusion that chromosomal changes affecting 21q22 might be primarily related to AR formation, whereas, changes of 8q22 produce the characteristic differentiation pattern leading to M2 morphology, consistently found in AML with t(8;21)(q22;q22). With regard to numerical abnormalities, -7 and +8 occurred about equally often in the two groups, whereas, +11 and -Y, especially when present as the sole aberrations, were predominantly found in AR+ patients. In contrast, -5 and +21 were exclusively found in AR- patients. The results indicate that AR+ AML is characterized by a relatively limited number of chromosomal abnormalities that are different from the aberrations found in AR- patients. This feature has not been emphasized in previous studies correlating hematologic and cytogenetic findings in AML. PMID- 3040222 TI - Polyclonal chromosomal evolution in a benign mixed salivary gland tumor. AB - Banding analyses of a human benign pleomorphic adenoma in the parotid gland revealed a polyclonal pattern where structural rearrangements predominated. These deviations were different from the anomalies previously observed in 100 mixed tumors. The reason found for the differences in all likelihood was x-ray treatment of tuberculous lymphadenitis in the neck during childhood. Implications regarding origin and development of pleomorphic adenomas are discussed. PMID- 3040223 TI - Familial polyposis coli and neurofibromatosis in the same patient: a family study. AB - This report describes a family wherein two major genes--one for familial polyposis coli and the second for von Recklinghausen's neurofibromatosis--were segregating. A patient whose father had familial poliposis coli and whose mother had neurofibromatosis manifested both of these disorders. PMID- 3040224 TI - Genetic resistance to mammary tumorigenesis in a mouse strain with high murine mammary tumor virus expression. AB - Although II-TES mice release large amounts of murine mammary tumor virus (MMTV) in milk, they are resistant to mammary tumorigenesis. High mammary tumor incidence was observed in (BALB/ca X II-TES)F1 and (C57BL/6N X II-TES)F1, whereas no mammary tumors developed in BALB/ca X OZ-F)F1. Mammary tumors developed in 68% of (OZ-F X (OZ-F X II-TES)F1 and 45% of (II-TES X (OZ-F X II-TES)F1). These results suggest that the II-TES mouse carries a recessive gene for mammary tumor resistance which does not inhibit MMTV release, and two independent dominant mammary tumor promoting genes which are inhibited by the resistant gene. PMID- 3040225 TI - Second stage tumor promoters: differences in biological potency and phorbol ester receptor affinity in C6 cells. AB - We have shown that the second stage tumor promoters mezerein (MEZ) and phorbol 12 retinoate 13-acetate (PRA) inhibit the gluccocorticoid-induced increase in glycerol phosphate dehydrogenase (GPDH) activity in C6 rat glioma cells with ED 50-values of 3.9 and 2.9 nM, respectively. Phorbol 12-myristate 13-acetate (PMA) was 10-fold less potent. MEZ was likewise more potent than PMA for inhibition of cAMP formation in response to isoproterenol. Binding competition studies using [3H]phorbol 12,13-dibutyrate ([3H]PDBu) yielded apparent Ki-values for MEZ and PRA of 50-70 nM. The large difference between the biological potencies of MEZ and PRA and their affinity for the major phorbol ester receptor suggest they may be acting through a more complicated mechanism in these cells. PMID- 3040226 TI - Induction of asynchronous replication of polyoma DNA in rat cells by ultraviolet irradiation and the effects of various inhibitors. AB - The ability of various DNA damaging agents to induce asynchronous replication of polyoma DNA (APR) in rat cells carrying integrated copies of these DNA sequences may provide a useful model for understanding mechanisms of gene amplification. The present study has explored in detail the ability of UV irradiation to induce APR in the polyoma transformed rat fibroblast cell line H3. We have found that the optimum condition for induction of APR was obtained by irradiating the H3 cells with UV-C (wavelength, 254 nm) at 1-2 J/m2. Irradiation with UV-B (270-360 nm) was much less effective, and no induction of APR was obtained with even high doses of UV-A (345-440 nm). This action spectrum provides evidence that the critical target for induction of APR is DNA. We found that when normal rat fibroblasts were irradiated with UV-C and then fused to H3 cells, this also led to induction of APR. These results provide evidence that the induction of APR by UV-C is mediated by a trans-acting factor. The induction of APR by UV-C was inhibited by high doses of cycloheximide or actinomycin D, suggesting that the production of this trans-acting factor requires de novo protein and RNA synthesis. On the other hand, low doses of cycloheximide or actinomycin D alone were able to induce APR, perhaps by blocking the synthesis of cellular factors that normally inhibit APR. Thus, induction of APR by UV-C provides a useful system for identifying cellular factors that might mediate or prevent the asynchronous replication of various DNA sequences. PMID- 3040227 TI - Role of free radicals in an adriamycin-resistant human small cell lung cancer cell line. AB - In two Adriamycin (Adr) resistant sublines (GLC4-Adr1 and GLC4-Adr2) of a human small cell lung carcinoma cell line, GLC4, cross-resistance for radiation was found. GLC4-Adr1 has an acquired Adr resistance factor of 44 after culturing without Adr for 20 days and GLC4-Adr2, the same subline cultured without Adr for 3 months, has a decreased but stable resistance factor of 8. One of the assumed mechanisms of Adr is that the effect is mediated through the formation of free radicals. Therefore free radical scavenging might play a role in these Adr resistant cell lines. Adr, H2O2, and X-ray induced cytotoxicity were evaluated. Glutathione (GSH) levels and activities of associated enzymes were determined as well as Adr, H2O2, and X-ray induced DNA breaks and repair. GSH level was decreased in GLC4-Adr1, but restored to the normal level in GLC4-Adr2. Superoxide dismutase, catalase, glutathione-peroxidase, and glutathione S-transferase were not elevated in the resistant sublines. Adr induced a decreased amount of DNA breaks in GLC4-Adr1 compared to GLC4. For X-ray and H2O2 a comparable amount of DNA damage was found. GLC4-Adr1 was able to repair DNA breaks induced by Adr, X ray, and H2O2 better than GLC4. In conclusion, no increased enzyme capacity for detoxification of free radicals could be detected in the cytosol of the resistant cells. The resistance against free radicals in the GLC4-Adr1 line may at least in part be a result of increased DNA repair. PMID- 3040228 TI - A-ring substituted estrogens as inhibitors of the MXT transplantable mammary ductal carcinoma. AB - A-ring substituted estrogens have been examined as growth inhibitors of the hormone dependent MXT murine mammary tumor. Certain of these estrogen analogues inhibited the growth of newly implanted as well as established MXT tumors when administered either by s.c. or i.p. injections or by intubation. These compounds were nontoxic over a broad range of active levels. Amino and nitro groups, introduced at position-4 of estrone 3-methyl ether were particularly carcinostatic, a property not shared by 4-bromoestrone 3-methyl ether. In addition tumor inhibition was greatly diminished by placing the nitro group at the other ortho position (i.e., carbon-2). Evidence indicates that the A-ring substituted estrogens may function as growth inhibitors via the estrogen receptor mechanism in the case of 4-nitro- and 4-aminoestrone. The 3-methyl ethers of these compounds also blocked tumor growth, possibly through in vivo dealkylation leading to the free phenolic A-ring substituted estrogens. On the other hand, A ring substituted 3-deoxyestrogens (particularly 4-nitro- and 4-aminoestratrien-17 beta-ol), which do not bind to receptor, were also excellent inhibitors of hormone dependent MXT breast tumors and therefore must express their activity by mechanisms other than that mediated by receptor. The A-ring substituted estrogens are unlike tamoxifen and diethylstilbestrol which (a) display toxicity at optimum inhibitory doses and (b) are inactive or marginally active in rodent breast cancer models. PMID- 3040229 TI - Correlation of V-src gene amplification with the tumorigenic phenotype in a Syrian hamster embryo cell line. AB - A preneoplastic cell line (10W) isolated after treatment of Syrian hamster embryo cells with asbestos was cotransfected with pSV2-neo DNA and Rous sarcoma virus DNA. Six of these colonies contained v-src DNA; however, none of the six initially expressed v-src RNA. Five of the clones failed to grow in soft agar (frequency, less than 10(-6)). One clone (61) grew in soft agar, but with a low frequency. Three of the clones (41, 61, and 62) were tumorigenic in nude mice and three were nontumorigenic. Cells cloned from soft agar or established from tumor explants expressed the v-src gene. The gene copy number of v-src, which was three to 10 in the original neoR clones, was increased approximately 10-fold in the soft agar-derived cell clones and tumor-derived cell lines. Cytogenetic analyses indicated that cells with amplified v-src contained double minute chromosomes. The results suggest that gene amplification influences the expression of the transfected oncogene and is a mechanism which can overcome the initial suppression of transcription of the v-src oncogene in the 10W cell line. PMID- 3040231 TI - Effects of recombinant human tumor necrosis factor on highly enriched hematopoietic progenitor cell populations from normal human bone marrow and peripheral blood and bone marrow from patients with chronic myeloid leukemia. AB - Previous studies using unseparated normal human bone marrow cells have indicated that recombinant tumor necrosis factor alpha (rTNF-alpha) can inhibit the in vitro colony growth by normal granulocyte/macrophage (CFU-GM) and erythroid (BFU E) progenitor cells in a dose-dependent manner. In the present studies, by using very low numbers of highly enriched normal bone marrow progenitor cell populations as target cells, we have extended these previous findings to provide convincing evidence that erythroid and myeloid colony growth suppression by rTNF alpha is manifested by a direct interaction between rTNF-alpha and CFU-GM and BFU E progenitor cells. In addition, the sensitivity of normal peripheral blood and chronic myeloid leukemia bone marrow CFU-GM and BFU-E colony growth to inhibition by rTNF-alpha was examined and found to be comparable with that of normal bone marrow CFU-GM and BFU-E. Although the continuous presence of high doses of rTNF alpha (5000 units/ml) was required in methylcellulose cultures for maximal CFU-GM (90%) and BFU-E (70%) colony suppression, short-term exposure (24 to 72 hr) of normal bone marrow-enriched progenitor cells to rTNF-alpha, in the absence of hematopoietic growth factors, was sufficient to irreversibly suppress up to 50 to 65% of CFU-GM colony growth. In contrast, the number of BFU-E colonies was increased under these conditions. If, however, hematopoietic growth factors (Mo-T cell-conditioned medium and erythropoietin) were present during preincubation of the cells with rTNF-alpha, BFU-E were then slightly suppressed while the extent of CFU-GM inhibition remained essentially the same. The suppressive effect of rTNF-alpha on erythroid and myeloid progenitor cell growth appears to be most pronounced on the more primative stages of committed progenitor cell development, since inhibition of CFU-GM- and BFU-E-derived colony growth progressively decreased with the delayed addition of rTNF-alpha to methylcellulose cultures. [3H]Thymidine incorporation was also inhibited by rTNF-alpha in normal bone marrow-enriched progenitor cell populations stimulated to proliferate in liquid culture by colony-stimulating factors. This effect was transient, however, since the activity of rTNF-alpha declined after the first 24 h of culture at 37 degrees C, particularly at low doses of rTNF-alpha where the activity was completely lost after 48 h of culture. This loss of activity appeared to be due to a decreased sensitivity of progenitor cells to the antiproliferative effects of tumor necrosis factor (TNF) after an initial exposure rather than a lack of available TNF.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3040232 TI - Mevalonic acid products as mediators of cell proliferation in simian virus 40 transformed 3T3 cells. AB - Effects of treatment with serum-free medium and 25-hydroxycholesterol (25-OH) on the cell cycle of simian virus 40-transformed 3T3 fibroblasts, designated SV-3T3 cells, were studied and compared with simultaneous effects on the activity of 3 hydroxy-3-methylglutaryl (HMG) CoA reductase and incorporation of [3H]mevalonic acid into cholesterol, Coenzyme Q, and dolichol. The data confirm our previous finding (O. Larsson and A. Zetterberg, Cancer Res., 46: 1233-1239, 1986) that 25 OH inhibits the cell cycle traverse of SV-3T3 cells specifically in early G1. In contrast, treatment with serum-free medium had no effect on cell cycle progression. The effect of 25-OH on the cell cycle traverse was correlated to a substantial decrease in the activity of HMG CoA reductase, whereas there was no change in the rate of [3H]mevalonic acid incorporated into cholesterol, Coenzyme Q, and dolichol. When the cells were exposed to serum-free medium, there was no depression of activity of HMG CoA reductase, and the rate of [3H]mevalonic acid incorporated into dolichol and cholesterol was not affected in any appreciable degree. In contrast the rate of Coenzyme Q synthesis was substantially decreased as a result of serum depletion. A similar decrease in Coenzyme Q synthesis was also achieved by treating the cells with cholesterol-poor serum. This indicates that the rate of Coenzyme Q synthesis is dependent on the concentration of cholesterol in the culture medium. In order to analyze whether some of the products in the mevalonic acid biosynthetic pathway may be of importance in the control of G1 traverse and cell proliferation of SV-3T3 cells, cholesterol, Coenzyme Q, and dolichol were added as supplements to cells treated with 25-OH. It was shown that dolichol was capable of overcoming the 25-OH-induced inhibition of G1 traverse efficiently, whereas cholesterol and Coenzyme Q were considerably less effective. Considered together with the fact that the activity of HMG CoA reductase and incorporation of mevalonic acid into dolichol were unaffected following serum-free treatment, the results suggest that maintenance of a certain level of de novo synthesis of dolichol may contribute to the capability of SV-3T3 cells to proliferate in serum-free medium. PMID- 3040230 TI - Chemiluminescence and oxygen radical generation by Walker carcinosarcoma cells following chemotactic stimulation. AB - The peptide N-formyl-Met-Leu-Phe stimulates chemotaxis and metastasis in rat Walker carcinosarcoma cells by a receptor-mediated pathway. Since oxygen radical generation follows chemotactic stimulation in leukocytes, we looked for similar responses in the Walker tumor. Upon incubation with 10(-6) M N-formyl-Met-Leu Phe, Walker cells elicited chemiluminescence in the presence of 5 X 10(-5) M luminol. The response peaked within 2 min and was maintained for greater than 20 min; it was dose dependent with a 50% maximal effective dose (ED50) value of 4.5 X 10(-8) comparable to the 50% maximal effective dose value for chemotaxis. The responses were significantly reduced but not abolished in the absence of calcium in the external medium and were elicited by the ionophore A23187. The lipoxygenase inhibitor nordihydroguaiaretic acid had almost no effect in decreasing the response, while flurbiprofen, a cyclooxygenase inhibitor was very effective at 10(-6) M. Evidence for the generation of oxygen radicals included: (a) marked inhibition of light emission in the absence of oxygen; (b) inhibition in the presence of superoxide dismutase, catalase, and mannitol; and (c) dose dependent reduction of acetylated cytochrome c. We postulate that activation of circulating tumor cells may facilitate metastasis by the release of toxic oxygen species. PMID- 3040233 TI - Ah receptor in human placenta: stabilization by molybdate and characterization of binding of 2,3,7,8-tetrachlorodibenzo-p-dioxin, 3-methylcholanthrene, and benzo(a)pyrene. AB - Aryl hydrocarbon hydroxylase (AHH, cytochrome P1-450) is highly inducible in several human cells and tissues exposed to specific halogenated and nonhalogenated aromatic chemicals of the "3-methylcholanthrene-type." In laboratory animals AHH induction is known to be regulated by binding of inducers to the Ah receptor, a soluble intracellular protein. However, the induction mechanism in the human species is incompletely understood largely because the Ah receptor, which seems to be essential to the induction process, has not previously been detectable in certain human cells and tissues (including placenta) that are highly responsive to AHH induction. We found that human placenta contains high concentrations of Ah receptor (comparable to the receptor concentrations in rat and mouse liver) but that special modifications were necessary in the assay techniques in order to detect and accurately quantitate receptor binding. Receptor was detected at concentrations approximately equal to 100 fmol/mg cytosol protein using [3H]2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) as the radioligand. This high concentration of specific binding sites was present only if the placental tissue was initially homogenized in a buffer containing sodium molybdate (10 or 20 mM). Without molybdate in the homogenizing buffer, specific [3H]TCDD binding was only about 35 fmol/mg. Specific Ah receptor binding also was detectable with [3H]-3-methylcholanthrene and, to a lesser extent, with [3H]-benzo(alpha)pyrene. The receptor sedimented near 9S on sucrose gradients whether molybdate was present or not. About 80% of specific binding was lost if excessive charcoal was used to adsorb "nonspecifically bound" ligand from cytosol prior to gradient analyses. The apparent affinity with which [3H]TCDD bound to Ah receptor in human placental cytosol was relatively low (apparent Kd approximately equal to 5 to 8 nM) when compared with the affinity of [3H]TCDD binding in rat or mouse hepatic cytosols (Kd approximately equal to 1 to 3 nM). These data suggest that while molybdate has very little effect on the quantity or molecular size of the rodent Ah receptor assay, it is very important in stabilizing the human Ah receptor. Our experiments demonstrate that human placenta contains a high concentration of Ah receptor and suggest that AHH induction in placenta is mediated through a receptor mechanism analogous to that previously established in tissues and cells from laboratory animals. PMID- 3040234 TI - Characterization of a human osteosarcoma cell line (Saos-2) with osteoblastic properties. AB - This study examines the osteoblastic properties of the established human osteosarcoma cell line Saos-2. Saos-2 cells inoculated into diffusion chambers, which were implanted i.p. into nude mice, produced mineralized matrix in 4 of 6 chambers at 8 weeks. In 5 of 6 chambers there was a strong positive alkaline phosphatase reaction. In culture the alkaline phosphatase levels increased with time and cell density, reaching very high levels at confluence: 4-7 mumol/mg protein/min. The cells show a sensitive adenylate cyclase response to parathyroid hormone, 50% effective dose = 2.8 nM, which increases with cell density and is further raised by dexamethasone treatment. They also exhibit typical binding of 1 25-dihydroxyvitamin D3 to 3.2S receptor protein with an apparent Kd of 0.21 nM; the numbers of sites per cell were 3,300 at 50,000 cells/cm2 and 1,800 at 280,000 cells/cm2. The presence of osteonectin was visualized with a monoclonal antibody which revealed a reticular pattern on the cell surface. Osteonectin was also detected in the medium by Western blots, migrating at around Mr 40,000 in nonreduced gels and Mr 44,000 in reduced gels. The Saos-2 cells thus possess several osteoblastic features and could be useful as a permanent line of human osteoblast-like cells and as a source of bone-related molecules. PMID- 3040235 TI - Changing incidence of hepatocellular carcinoma in Japan. AB - A trend in the incidence of hepatocellular carcinoma (HCC) in Japan was studied from the data of the Osaka Cancer Registry (population, 8,512,351 in 1981) for the period of 1963-1983, the Vital Statistics of Japan, Ministry of Health and Welfare, and the Japan Autopsy Registry which contained 594,132 individually filed cases in the 26-year period from 1958 to 1983. Both cancer registry data and autopsy records showed a more than 2-fold increase in HCC incidence, particularly in the last 10 years or so, among males and a less pronounced increase in females. The same trend was borne out by the cancer registries of Nagasaki City and Miyagi Prefecture and the Vital Statistics. When studied with the autopsy data, it was found that the numbers of autopsies for cirrhosis without HCC and autopsies for HCC (with and without cirrhosis) were about the same in 1958-1961 and that currently (1980-1983) the latter is about 2 times the former. As one of the possible causes of increase in HCC incidence other than prolonged survival of patients with cirrhosis, chronic non-A, non-B hepatitis is discussed. PMID- 3040236 TI - Differential responsiveness of normal and simian virus 40-transformed BALB/c 3T3 cells to retinoic acid: rapid enhancement of epidermal growth factor receptor binding in a simian virus 40-3T3 variant. AB - The effects of retinoic acid on the epidermal growth factor (EGF) receptor binding and cell growth of normal and simian virus 40 (SV40)-transformed BALB/c 3T3 cells were compared under identical culture conditions. Retinoic acid induced a rapid enhancement of EGF binding to SV40-transformed cells. Half-maximal enhancement occurred at about 7 h after the cells were exposed to 20 ng/ml of retinoic acid, and maximal stimulation (from 2.5- to 3.5-fold over the control) was obtained after 12 h of exposure. The kd of the control and retinoic acid treated cells was calculated to be 8.0 X 10(-10) M and 8.2 X -10 M, respectively. However, the number of unoccupied EGF binding sites increased from 0.98 X 10(4) to 2.28 X 10(4) per cell. Normal 3T3 cells would not respond to retinoic acid unless they were cultured in serum-containing medium. After 96 h of exposure, only a 50% enhancement of EGF binding was observed. The EGF receptor number of the untreated normal cells was calculated to be 1.82 X 10(4) per cell, twice the number expressed by untreated SV40-transformed cells. The increase of EGF receptor number caused by retinoic acid in SV40-transformed cells was blocked by either actinomycin D or cycloheximide treatment. These results indicated that SV40 transformation of BALB/c 3T3 cells altered the regulatory mechanism governing the complement of cell surface EGF receptors. PMID- 3040237 TI - Markedly different antibody responses to immunized small cell and non-small cell lung cancer cells. AB - Monoclonal antibodies (MoAbs) to antigens of human small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC) were produced from BALB/c mice immunized with either intact cultured cells or membrane preparations from cultured tumor cells. Of 172 MoAbs produced from two NSCLC immunized mice and reactive to NSCLC cells, 137 bound staphylococcal Protein A directly, and only 11 of these 172 MoAbs were significantly reactive with cultured SCLC cells by a solid-phase radioimmunoassay. In contrast, only 16 of 99 MoAbs produced from two SCLC immunized mice and reactive to SCLC cells directly bound Protein A, and most were of an IgM isotype, but 98 of 99 of these antibodies also reacted with cultured NSCLC target plates. Twenty hybridomas producing antibodies reactive with lung cancer cells but not with a B-lymphoblastoid line were cloned. Eleven of these cloned hybridomas were from NSCLC fusions, and nine were from SCLC fusions. When representative hybridoma supernatants were tested against a broad panel of SCLC and NSCLC target plates a similar pattern was seen, with the supernatants from NSCLC-derived hybridomas only reacting strongly to the NSCLC target plates, but the supernatants from SCLC-derived hybridomas always reacting to both SCLC and NSCLC plates. We conclude that the humoral response to immunization with NSCLC cells or membrane preparations is predominantly IgG and that to SCLC is predominantly IgM. Furthermore, many IgG MoAbs reactive with NSCLC lines are poorly reactive with SCLC cells. PMID- 3040238 TI - Formation of blocking lesions at identical DNA sequences by the nitrosourea and platinum classes of anticancer drugs. AB - cis-Diamminedichloroplatinum (II) (cisplatin) compounds and the chloroethylnitrosoureas are two different classes of anticancer drugs that work by modifying DNA covalently. We have compared the platinating drug cisplatin with the alkylating drug bischloroethylnitrosourea and other chloroethylnitrosoureas by modifying double stranded DNA in vitro and identifying blocking lesions that impede the progress of Escherichia coli DNA polymerase. Despite their very different structures and reactivities, cisplatin and the chloroethylnitrosoureas from primary blocking lesions at identical sequences, those containing adjacent guanosines on the same DNA strand. In tumor virus SV 40 DNA, a very strong target for both types of drugs is the regulatory sequence GGGCGG, which is repeated six times and is an important sequence for viral replication and an essential sequence for expression of the viral transforming gene. Sequences related to these GC box elements are known to be present in the flanking regions of many retroviruses and oncogenes, thus raising the possibility that the targeting of these sequences in tumor cells contributes to drug activity. PMID- 3040240 TI - Expression of neurotransmitter receptors and myc protooncogenes in subclones of a human neuroblastoma cell line. AB - Phenotypic variability of the human neuroblastoma cell line SK-N-SH was studied with the use of three subclones that interconvert at a slower rate than the parent cell line, i.e., a neuroblast-type subclone (SH-SY5Y), a nonneuronal, strongly substrate adherent subclone (SH-EP), and an intermediate type subclone (SH-IN). Rhodamine-phalloidin staining of actin fibers revealed differences in the cytoskeleton morphology of the three subclones, while the clathrin subunit proteins (heavy and light chains), components of coated vesicles, were invariant. Dramatic differences were observed for the expression of neurotransmitter systems, i.e., the mu and delta opioid receptor, the muscarinic cholinergic receptor and its effect on phosphatidylinositol turnover, and the uptake1 transporter for catecholamines. While these systems were strongly expressed in the parent line and the neuroblast-like clones SH-SY5Y and SH-IN, they were absent or barely detectable in the nonneuronal EP clone. Furthermore, the protooncogenes N- and c-myc were only expressed in the neuroblast containing lines, consistent with their growth characteristics of fully transformed cells. The strong c-myc expression in the absence of c- or N-myc amplification in SK-N SH, adds a new form of high protooncogene activity in neuroblastoma cell lines. The remarkable differences of neurotransmitter systems and myc expression among the various phenotypes of human neuroblastoma cells should be considered in the therapy of neuroblastoma. PMID- 3040239 TI - Quantitative structure-activity relationships: analysis of interactions of 2,3,7,8-tetrachlorodibenzo-p-dioxin and 2-substituted analogues with rat, mouse, guinea pig, and hamster cytosolic receptor. AB - The competitive receptor binding affinities of thirteen 2-substituted 3,7,8 trichlorodibenzo-p-dioxins to hepatic cytosol from rat, mouse, guinea pig, and hamster were determined with [3H]-2,3,7,8-tetrachlorodibenzo-p-dioxin as the radioligand. Multiple parameter linear regression analysis of the binding data from the four species gave the following equations: pEC50 (rat) = 7.196 + 0.600 pi - 0.255 delta Es - 1.683 HB pEC50 (mouse) = 6.365 + 1.641 pi + 1.206 sigma 0 pEC50 (hamster) = 7.416 + 1.026 pi + 0.509 delta Es + 0.748 sigma 0 pEC50 (guinea pig) = 6.892 + 1.035 pi where pi, delta Es, HB, sigma 0, and Vw are physicochemical parameters for substituent lipophilicity, steric effect, hydrogen bonding capacity, electronegativity, and van der Waals volume (relative to H), respectively. These equations demonstrate that there are important species differences in the receptor protein binding site interactions with the substituted analogues. These data, coupled with the known species differences in the molecular properties of the receptor proteins, are evidence for a heterologous nature of the receptor between mammalian species. Multiple parameter linear regression analysis of the relative aryl hydrocarbon hydroxylase (AHH) induction potencies of these analogues in rat hepatoma H-4-II E-cells in culture gave the following equation. The correlation pEC50 (AHH induction) = 3.208 + 0.950 pEC50 (rat binding) - 0.955 delta B5 between receptor binding and AHH induction was dependent on a steric parameter (delta B5, STERIMOL) and the results suggest that an additional substituent-dependent process (e.g., an activation step) may be required after initial ligand-receptor binding for the ultimate expression of the receptor-mediated response (i.e., AHH induction). PMID- 3040241 TI - Some emerging general principles in the pathogenesis of hepatocellular carcinoma. AB - A rational concept of the key steps in the multistep process of cancer development in liver is emerging. This concept proposes that the strategy of cancer development consists of two major sequences: (a) the genesis of persistent benign focal proliferations (clonal nodules) and (b) the development of hepatocellular cancer from one or more such nodules. Sequence (a) comprises the classical 'initiation and promotion' of chemical carcinogenesis. Sequence (b) is dominated by persistent cell proliferation. In the precancerous steps, cell proliferation is almost balanced by cell loss, leading to only a slow increase in the size of the nodules. With cancer, this balance is lost, leading to a much more rapid enlargement of the focal lesion. The carcinogenic process in the liver is viewed initially as a form of physiological adaptation to certain types of xenobiotic agents generating new focal cell populations. The animals with such new focal lesions are much better able to resist the toxic or lethal effects of many environmental hazards. According to this view, liver cancer development is a consequence of a derivative of the basic adaptation process. PMID- 3040242 TI - The development of hepatocellular cancer in humans. AB - The biological characteristics of hepatocellular cancer vary appreciably in different parts of the world, but especially between regions with very high and low incidences of the tumour. Hepatocellular cancer is multifactorial in origin, and the pattern of its aetiological associations differs between populations at high and low risk. In Africans and Chinese, who have the highest incidences of hepatocellular cancer, the hepatitis B virus is the most important causal association. The viral carrier state is acquired during early childhood, and carries a relative risk for the development of the tumour of over 200. Integration of hepatitis B virus DNA probably acts as a genotoxic initiator in the multistep process of hepatocarcinogenesis, although the precise mechanisms involved have not been determined. Aflatoxin ingestion may also have an aetiological role in high incidence regions, probably as a genotoxic or epigenetic promoter to hepatitis B virus-initiated carcinogenesis. In low risk populations cirrhosis is the most important causal association of hepatocellular cancer. The cirrhosis is often the result of alcohol abuse, but the tumour may complicate all aetiological forms of this disease. Whether neoplasia is an inevitable consequence of the hyperplasia of cirrhosis, or the increased hepatocyte turnover rate acts as a promoter is not known. Hepatitis B virus infection plays a lesser part, and aflatoxin no part at all. PMID- 3040243 TI - Interactive effects of chemical carcinogens and hepatitis B virus in the pathogenesis of hepatocellular carcinoma. AB - Liver cancer is one of the most prevalent forms of cancer in the world. Hepatitis B virus (HBV) is considered to be a major aetiological factor. Evidence from epidemiological studies has also indicated that environmental contaminants such as mycotoxins may, either in combination with HBV or independently, be important aetiological factors in the pathogenesis of primary hepatocellular carcinoma (PHC). Laboratory data also suggest an interplay between viral and chemical factors in the multifactorial aetiology of PHC. Aflatoxin B1, the chemical carcinogen most frequently implicated in the aetiology of hepatocellular carcinoma is a procarcinogen that must be activated by mixed-function oxidases to an electrophilic metabolite before it can exert its carcinogenic effects. Interindividual differences (greater than 10-fold) in the metabolic activation of aflatoxin B1 are observed. These differences may play a part in an individual's oncogenic susceptibility to aflatoxin B1. Chemical carcinogens and integrated HBV may activate cellular oncogenes, eg N-ras, and inactivate tumour suppressor genes. Recently developed methods that allow monitoring of aflatoxin B1 and HBV exposures and also genetic damage caused by these agents in individuals should help in biochemical and molecular epidemiological studies concerning the aetiology of hepatocellular carcinoma. We identify areas of uncertainties and of future experimentation and propose a hypothesis of liver carcinogenesis. PMID- 3040244 TI - Prevention of primary liver cancer by vaccination. PMID- 3040245 TI - Combination chemotherapy with ifosfamide, mitomycin, and cisplatin in advanced non-small cell lung cancer. AB - Thirty-two evaluable patients with stage III non-small cell lung cancer were treated with the combination of ifosfamide (4 g/m2), with uroprotective mesna, mitomycin (6 mg/m2), and cisplatin (100 mg/m2). The overall response rate was 68.8% (complete response rate, 21.9%; partial response rate, 46.9%). Toxicity was moderate and manageable. Based on the response rate observed, this three-drug combination could be considered for future randomized phase III trials against non-small cell lung cancer. PMID- 3040247 TI - Phase II evaluation of iproplatin in refractory germ cell tumors: a Southeastern Cancer Study Group trial. PMID- 3040246 TI - Phase II trial of 4'-deoxydoxorubicin in advanced non-small cell lung cancer. PMID- 3040248 TI - Subarachnoid hemorrhage from brain tumors in childhood. AB - Six children are reported in whom subarachnoid hemorrhage was an initial symptom of brain tumor. In our neurosurgical clinics, this represented 3.6% of pediatric brain tumors and showed a frequency equal to aneurysmal rupture among nontraumatic subarachnoid hemorrhage of children. In pediatric patients, hemorrhages from brain tumors occur predominantly in the posterior fossa. The medulloblastoma, which had been believed to bleed rarely, is now realized to be a common source of tumor hemorrhages in such cases. The introduction of CT scan facilitates early recognition of hemorrhagic stroke from brain tumors and prompt management for acute intracranial hypertension and brainstem dysfunction. Although the patients achieve favorable recovery from their initial catastrophic condition, the ultimate prognosis, in the majority of cases, is still rather poor because such hemorrhages usually develop from a malignant tumor. The present and other recent reports indicate that the incidence of hemorrhagic stroke from brain tumors in pediatric patients is much higher than has been thought and is an important cause of subarachnoid hemorrhage in this age group. PMID- 3040249 TI - Outcome of children with primary intramedullary spinal cord tumors. AB - The influence of clinical and treatment factors on the outcome of children with primary intramedullary spinal cord tumors (PST) was evaluated by reviewing the records of 26 children diagnosed during the 15-year period 1970-1984. Five-year survival was 39%, but 5-year event-free survival (EFS) was only 14%. Eighteen month EFS was 53% (9/17) among children with low-grade astrocytoma. 100% (2/2) with ependymoma, and 0 of 7 with anaplastic astrocytoma or ganglioglioma. There was no significant difference in the 18-month EFS by location of tumor, duration of symptoms, or extent of surgical removal. Five of 9 children with locally recurrent PST had a second operation, and 4 were alive a median of 56 months later. PST disseminated to the leptomeninges or the III ventricle in 5 children: 2 at diagnosis, 2 as the first sign of disease relapse, and 1 after local recurrence. Given the poor outcome of our children, different methods of treatment for children with tumors in this location should be evaluated. PMID- 3040250 TI - A new catalyst for reductive cleavage of methylated glycans. AB - Several per-O-methylated D-glucans and D-fructans were used as models in an attempt to identify new catalysts for carrying out reductive cleavage. Included in these model studies were several D-glucans that contained 4-linked D glucopyranosyl residues as well as one having a 4-linked D-glucitol residue, as both types of residue had previously been found to give rise to substantial proportions of artifactual products. These studies led to the development of a new catalyst for carrying out reductive cleavage, namely, a mixture of 5 equivalents of trimethylsilyl methanesulfonate (Me3SiOSO2Me) and 1 equivalent of boron trifluoride etherate (BF3 . Et2O) per equivalent of acetal. This new catalyst was found to accomplish the reductive cleavage of per-O-methylated, 4 linked D-glucopyranosyl residues and 4-linked D-glucitol residues, to give only the expected derivatives of 1,5-anhydro-D-glucitol and D-glucitol, respectively. The mixture of Me3SiOSO2Me and BF3 . Et2O also catalyzed reductive cleavage of the D-fructofuranosyl residues of per-O-methylated sucrose and inulin, to give only the expected derivatives of 2,5-anhydro-D-mannitol and 2,5-anhydro-D glucitol. Indeed, when used alone, Me3SiOSO2Me also rapidly catalyzed the reductive cleavage of D-fructofuranosyl residues, but, under the same conditions, D-glucopyranosyl residues were unaffected. The results of these and other model studies demonstrated that catalysis of reductive cleavage by the mixture of Me3SiOSO2Me and BF3 . Et2O occurs in a synergistic manner. Examination of the mixture of Me3SiOSO2Me and BF3 . Et2O by 1H-n.m.r. spectroscopy demonstrated that a reaction occurs to generate trimethylsily fluoride and species of the type F2BOSO2Me, FB(OSO2Me)2, or B(OSO2Me)3 via ligand exchange. PMID- 3040251 TI - Expression of endogenous lectins in human small-cell carcinoma and undifferentiated carcinoma of the lung. AB - Endogenous carbohydrate-binding proteins (lectins) were detected in specimens of tumor tissue (undifferentiated carcinoma and xenografted small-cell carcinoma) from human lung. Fractionation of salt and detergent extracts on different sets of Sepharose columns covalently derivatized with lactose, asialofetuin, melibiose, mannan, and fucose, successive elution with a chelating agent and a specific sugar, and analysis of the eluates by gel electrophoresis, resulted in the characterization of the profiles of endogenous carbohydrate-binding proteins. All preparations were devoid of enzymatic activity. Comparison between the patterns of the two types of lung carcinoma showed significant qualitative differences, e.g. the presence of fucose-binding proteins of apparent molecular weights 60,000 and 80,000 in the undifferentiated carcinoma, and the presence of beta-galactoside-binding proteins of apparent molecular weights 18,000 and 22,000 in the small-cell lung carcinoma. These proteins were not detectable in normal lung tissue. Such differences, documented for the first time for human lung tumors, are of potential importance as a step towards a lectin-based refinement of lung-cancer diagnosis and therapy. PMID- 3040252 TI - The duct system of the avian salt gland as a transporting epithelium: structure and morphometry in the duck Anas platyrhynchos. AB - The duct system of the nasal salt gland of the duck comprises central central canals, secondary ducts and main ducts. The secondary and main ducts consist of a layer of columnar cells overlying a layer of small cuboidal cells. The columnar cells have complex intercellular spaces showing evidence of Na+K+-ATPase at the apical regions. Approximately 70% of surface area of the duct system is external to the gland. During adaptation to salt water the duct system increases in size as does the gland. Although the components of the gland of adapted ducks, including the duct system within the gland, increase in size compared with normal ducks, the percentage volume densities of the components remain similar in both categories of ducks, i.e. the duct system increases in size in proportion to the glandular tissue. The volume of the duct system external to the gland is six to seven times larger than the volume within the gland. Thus, if ductal modification of secreted fluid occurs, it will be most likely to take place in the ducts external to the gland. Total surface areas of the duct system were measured from serial sections of glands and ducts from one normal and one adapted duck. These were used to calculate possible flux rates of water and sodium across the duct epithelium, assuming the occurrence of either water reabsorption of sodium secretion. Although these flux rates are high it is shown that they are similar to calculated flux rates across the luminal surface of the secretory tubules. PMID- 3040253 TI - In vivo binding and uptake of low-density lipoprotein-gold- and albumin-gold conjugates by parenchymal and sinusoidal cells of the fetal rat liver. AB - To elucidate the participation of fetal rat liver cells in the receptor-mediated internalization of low-density lipoproteins (LDL), rat fetuses were injected with either LDL-gold or albumin-gold conjugates. The degree of binding and uptake of LDL-gold and albumin-gold by parenchymal and sinusoidal cells of the fetal rat liver differs markedly. Endothelial cells exhibit low LDL-gold uptake. In contrast, parenchymal cells internalize LDL-gold more actively (45 +/- 8 LDL conjugates/100 micrometers2 cytoplasm within 60 min). Kupffer cells exceed this value by a factor of 20. The uptake of albumin-gold by endothelial and Kupffer cells is high, whereas it is extremely low in parenchymal cells. Estradiol pretreatment causes a significant doubling (p less than 0.05) of the LDL-gold particle density/100 micrometers2 cytoplasm both in parenchymal and Kupffer cells, whereas estradiol has no effect on the albumin uptake. The results strongly indicate that LDL uptake by parenchymal and Kupffer cells in the fetal rat liver is mediated by estrogen-inducible receptors, which may correspond to B, E receptors in the adult liver. PMID- 3040254 TI - A morphometric and ultrastructural study of lithium-induced changes in the medullary collecting ducts of the rat kidney. AB - Rats were given a lithium-containing diet (40 mmol/kg) to study the effect of lithium on the structure of collecting ducts from the inner stripe of the outer medulla. The results show that there is a significant increase in the volume density of collecting ducts already after one week on this diet. The volume density of both intercalated and principal cells increases, whereas the volume density of mitochondria in the cytoplasm increases in the intercalated cells only. The increased volume of both principal and intercalated cells seems to be part of a general hyperplasia and hyperactivity of the collecting duct, which may in some way be related to the effects of lithium on vasopressin-mediated mediated water transport. The specific changes in the intercalated cells may be a consequence of the effects of lithium on distal nephron potassium and hydrogen ion transport in the distal nephron. PMID- 3040256 TI - [The present status and methods of control of diarrhea]. PMID- 3040257 TI - [A survey of types and distribution of enterovirus among residents of Yantai City]. PMID- 3040255 TI - Origin of regenerating Leydig cells in the testis of the adult rat. An ultrastructural, morphometric and hormonal assay study. AB - Ethane dimethanesulphonate (EDS) was used as a specific cytotoxin to eliminate the Leydig cell population of the adult rat testis. Ultrastructural, morphometric and serum gonadotrophin and testosterone analysis was used to study the response of the intertubular tissue of the testis from 1 day to 10 weeks after EDS treatment. In control animals, the testis contained approximately 28 million Leydig cells and 8 million macrophages. Three to seven days after EDS treatment, Leydig cells were absent and serum testosterone was undetectable. Macrophage numbers increased three-fold by 3 days and returned to pretreatment values thereafter. At 2 and 3 weeks post-EDS, foetal-type Leydig cells (approximately 1 2 million per testis) appeared in proximity to perivascular and peritubular tissues, a feature also observed at 4 weeks when numerous such cells (approximately 15 million per testis) formed prominent clusters in perivascular and peritubular locations. Between 6 and 10 weeks after EDS treatment, the foetal type Leydig cells were transformed morphologically into adult-type Leydig cells, they occupied central intertubular positions and their numbers were restored to pretreatment values. Regeneration of Leydig cells was reflected by elevated serum testosterone levels which returned towards the normal range. The results demonstrate the regenerative capacity of the testicular intertubular tissue and indicate a dual site of origin of Leydig cells which initially resemble foetal type Leydig cells prior to establishing the adult-type Leydig cell population. The morphological pattern of Leydig cell regeneration suggests that in addition to gonadotrophic stimulation, local testicular factors from the seminiferous tubules may stimulate Leydig cell growth. PMID- 3040258 TI - Interactions between a DNA-binding transcription factor (COUP) and a non-DNA binding factor (S300-II). AB - We have identified previously two transcription factors, COUP (chicken ovalbumin upstream promoter) and S300-II, from HeLa cell nuclear extracts. In this paper, the purine base and the phosphate backbone contact sites for the COUP transcription factor were defined. These studies indicate that the COUP box transcription factor interacts with specific base residues in the major groove of the DNA helix. In addition, we have purified the S300-II factor over 100,000 fold. The polypeptide possessing functional transcriptional activity has been identified by SDS-PAGE followed by gel-slice elution and a renaturation assay. It is absolutely required for in vitro function of the ovalbumin promoter. In addition, S300-II stimulates transcription from the MMTV and lysozyme promoters. Kinetic studies probing the interaction of S300-II with COUP factor suggest that it may stabilize COUP-promoter complexes by slowing their rate of dissociation. PMID- 3040259 TI - Parental legacy determines methylation and expression of an autosomal transgene: a molecular mechanism for parental imprinting. AB - We have created a transgenic mouse strain in which an autosomal transgene bearing elements of the RSV LTR and a translocated c-myc gene obeys very unusual rules. If the transgene is inherited from the male parent, it is expressed in the heart and no other tissue. If it is inherited from the female parent, it is not expressed at all. This pattern of expression correlates precisely with a parentally imprinted methylation state evident in all tissues. Methylation of the transgene is acquired by its passage through the female parent and eliminated during gametogenesis in the male. These observations provide direct molecular evidence that autosomal gene expression can depend upon the sex of the parent from which the gene is inherited. They also provide a plausible mechanism for understanding parental imprinting that may be relevant to the failure of parthenogenesis in mammals, the apparent non-Mendelian behavior of some autosomal genes, and the role of methylation in gene regulation. PMID- 3040260 TI - Site-specific recombination intermediates trapped with suicide substrates. AB - A family of novel substrates was designed to enable the efficient accumulation of intermediates in site-specific recombination. Strategically placed nicks allow these "suicide substrates" to initiate the reaction but prevent its completion or reversal. Consequently, it has been possible to determine that lambda site specific recombination proceeds by a pair of sequential single-strand exchanges. These results rule out that class of models invoking a concerted cutting of all four DNA strands. The sequential strand exchanges are executed in a strictly prescribed order that is the same in both integrative and excisive recombination. This specified order appears to be governed by the arrangement of bound proteins distal to the sites of strand exchange. Furthermore, when provided with an appropriate 5' OH acceptor, the Integrase protein has the capacity to execute a single DNA strand transfer in a nonreciprocal reaction. PMID- 3040261 TI - The DNA-binding domains of the jun oncoprotein and the yeast GCN4 transcriptional activator protein are functionally homologous. AB - The jun oncoprotein, which causes sarcomas in chickens, and the DNA-binding domain of yeast GCN4, which coordinately regulates the expression of amino acid biosynthetic genes, show significant homology. In yeast cells deleted for the GCN4 gene, GCN4 function can be conferred by a hybrid protein in which the GCN4 DNA-binding domain is replaced by the homologous region of jun. Moreover, in strains containing various mutations of the GCN4 binding site in the HIS3 promoter, HIS3 expression is affected similarly by the hybrid protein and by GCN4. These results indicate that the jun oncoprotein binds the same DNA sequences as GCN4, and strongly suggest that jun is derived from a normal cellular transcription factor (possibly AP-1, which recognizes similar sequences). This provides direct evidence for the idea that alterations in the machinery for proper gene expression can lead to the oncogenic state. PMID- 3040262 TI - Positive and negative regulation of transcription in vitro: enhancer-binding protein AP-2 is inhibited by SV40 T antigen. AB - We have purified a 52 kd protein, AP-2, that binds to enhancer regions of SV40 and human metallothionein IIA (hMT IIA) and stimulates RNA synthesis from these promoters in vitro. Surprisingly, AP-2 also binds to two SV40 early promoter regions recognized by Sp1 and T antigen. Juxtaposed binding sites for AP-2 and Sp1 in the 21 bp repeats may facilitate productive interactions between the two factors. In contrast, sequence-specific binding of AP-2 to SV40 and hMT IIA DNA is inhibited by the viral repressor protein T antigen. Furthermore, T antigen inhibits AP-2-dependent transcriptional activation of the hMT IIA promoter in vitro. The inhibition is neither a direct nor an indirect result of T antigen binding to DNA, because the hMT IIA promoter lacks T antigen binding sites. Instead, sedimentation studies suggest that protein-protein interactions between AP-2 and T antigen block AP-2 binding to DNA. These findings suggest novel mechanisms for mediating positive and negative regulation of transcription. PMID- 3040263 TI - An antitermination protein engages the elongating transcription apparatus at a promoter-proximal recognition site. AB - As a transcriptional activator, the N protein of phage lambda acts to suppress transcription termination by recognizing a promoter-proximal site, nut, which is separated from the terminators by thousands of base pairs. We demonstrate here that N interacts with the elongating RNA polymerase in transit through the boxB domain of nut. This interaction leads to the stable association of N as an integral component of the transcription apparatus. During subsequent elongation, N translocates along with polymerase through several defined terminators positioned beyond nut. Therefore, by being an operon-specific subunit of the transcription apparatus, N presumably prevents the interaction of polymerase with termination signals. PMID- 3040264 TI - DNA topoisomerase II is required for condensation and separation of mitotic chromosomes in S. pombe. AB - We show that DNA topoisomerase II (topo II) is continuously required for mitotic chromosome changes in Schizosaccharomyces pombe. We constructed cold-sensitive (cs) or temperature-sensitive (ts) strains mutated in the genes coding for topo II (top2) and beta-tubulin (nda3). The ATP-dependent activity of the top2cs gene product is cs in vitro. The cloned top2cs gene sequence predicts an amino acid substitution. A cs top2-cs nda3 double mutant at 20 degrees C shows long, entangled chromosomes, which condense and separate upon the shift to permissive temperatures. If spindle formation is prevented at permissive temperatures, the chromosomes condense but do not separate. Thus topo II is required for final chromosome condensation; moreover, pulse-shift experiments show that topo II is required for chromatid disjuction. Experiments with ts top2-cs nda3 cells show that topo II is also required for chromosome separation in anaphase: inactivation of topo II and activation of beta-tubulin allow normal spindle formation but result in "streaked" chromosomes. PMID- 3040265 TI - Dual regulation of the yeast CDC28-p40 protein kinase complex: cell cycle, pheromone, and nutrient limitation effects. AB - A 40 kd polypeptide that coprecipitates with the CDC28 gene product in immune complexes is specifically phosphorylated by the CDC28 protein kinase. Using this reaction, we detect activity only in extracts from dividing G1 phase cells. Exit from G1 by entry into S phase or the preconjugatory state induced by mating pheromone correlates with loss of p40 phosphorylation activity. Inactive extracts from cdc28 mutants complement extracts from cells arrested in S or M phase, suggesting that non-G1 cells are deficient in an exchangeable activating factor. Stationary and pheromone-treated cultures are rich in this exchangeable factor, but possess an inactive kinase that is not activated by complementation. cAMP deficient mutants resemble stationary cells. PMID- 3040266 TI - A specific mismatch repair event protects mammalian cells from loss of 5 methylcytosine. AB - 5-Methylcytosine spontaneously deaminates to form thymine, thus generating G/T mispairs in DNA. We investigated the way in which these lesions are addressed in mammalian cells by introducing specific G/T mispairs into the genome of SV40 and determining the fate of the mismatched bases in simian cells. Mispairs were incorporated in 12 bp synthetic duplexes ligated into SV40 DNA between the BstXI and TaqI restriction sites. Analysis of 347 plaques obtained after transfection of this modified DNA indicated that mispairs were corrected in 343 cases (99%), revealing 314 repair events in favor of guanine (90%) and 29 in favor of thymine (8%). Correction in favor of guanine occurred regardless of the orientation of the mispair in DNA and regardless of whether the mispair was in the commonly methylated CpG dinucleotide. These results attest to a specific mismatch repair pathway that restores G/C pairs lost through deamination of 5-methylcytosine residues. PMID- 3040267 TI - Reversible silencing of enhancers by sequences derived from the human IFN-alpha promoter. AB - The virus-responsive element of the IFN-alpha 1 promoter, VRE(IFN alpha), comprises two imperfect 19 bp repeats, repA and repB. VRE(IFN alpha), tetrameric repA, and tetrameric GAAAGT (a subsequence of repB) or tetrameric AAGTGA conferred inducibility on a reporter gene when placed upstream of a complete or truncated promoter. Induced transcription was weak with a minimal promoter (TATA box only), but was strongly stimulated by the SV40 enhancer placed immediately upstream of the inducible element. Surprisingly, under noninduced conditions, tetrameric repA, GAAAGT, and AAGTGA (but not VRE(IFN alpha)) completely silenced enhancement of constitutive transcription by the SV40 72 bp repeat when interposed between the latter and the TATA box; silencing was fully abrogated by induction. PMID- 3040268 TI - A role for ID repetitive sequences in growth- and transformation-dependent regulation of gene expression in rat fibroblasts. AB - A set of mRNAs tagged by repetitive sequences of the ID family were found to accumulate following growth-factor-induced transition of normal (FR3T3) rat fibroblasts from a quiescent to a proliferative state. The levels of the same transcripts were also increased following transformation by polyoma virus and by ras and myc oncogenes. The presence of the ID element appeared to be determinant, since a similar pattern of expression was observed for a construct where one element had been inserted in the 3' noncoding region of a rabbit beta-globin gene expressed under control of an SV40 promoter. PMID- 3040269 TI - Conservation of a receptor/signal transduction system. PMID- 3040270 TI - Regulation of T-cell activation and T-cell growth factor (TCGF) production by hydrogen peroxide. AB - Activated macrophages are known to release a variety of immunoregulatory substances including the low-molecular-weight substances hydrogen peroxide and lactate. We report here that lactate but not hydrogen peroxide is capable of supporting a substantial production of T-cell growth factor (TCGF) in cultures of accessory cell-depleted splenic T-cell populations after stimulation with concanavalin A. Hydrogen peroxide and its biosynthetic precursor superoxide anion (O2-) mediate, however, a strong augmentation of the TCGF production by accessory cell-depleted T-cell populations in the presence of lactate. Lactate inhibits the incorporation of [3H]thymidine in short-term cultures (18-26 hr) of accessory cell-depleted T cells. This confirms the rule that (optimal) production of T-cell growth factor requires a growth inhibitory signal. Concentrations of hydrogen peroxide which augment TCGF production most effectively (i.e., 1 X 10(-5) M) do not inhibit the incorporation of [3H]thymidine; and higher concentrations (3 X 10(-5)-1 X 10(-4) M) of hydrogen peroxide inhibit both the production of TCGF and the incorporation of [3H]thymidine. In agreement with the augmenting effect of hydrogen peroxide on TCGF production, it was observed that the proliferative response in mixed lymphocyte cultures is suppressed by catalase and augmented by 1 X 10(-5) M H2O2. Proliferative and cytotoxic responses in mixed lymphocyte cultures with an external source of interleukin 2 (IL-2) in contrast, are not augmented by 1 X 10(-5) M H2O2. The relatively high concentration of 1 X 10(-4) M hydrogen peroxide was found to inhibit the proliferative responses in mixed lymphocyte cultures with or without external IL-2 but not the cytotoxic response in the presence of IL-2. This indicates that CTL precursor cells may be relatively resistant against H2O2. PMID- 3040271 TI - Morphological basis for multiple interactions of ethidium bromide (EB) with yeast Candida utilis. AB - Exposure of yeast cells to EB produced multiple effects on the cellular organelles: changes in the plasma membrane characterized by 75 to 110 nm deep pits; polymorphisms of the mitochondria ranging from cup-shaped, ring-shaped, ribbon-shaped, dumbbell-shaped structures to finally the formation of very elongated mitochondria (up to 4.5 micron in length); an increase in the length and number of endoplasmic reticulum; an increase in the number of cytoplasmic vesicles whose diameter varied between 25 to 45 nm. Furthermore, EB inhibited cytochrome c oxidase and cytochrome b biosynthesis, stimulated cytochrome c biosynthesis and uncoupled oxidative phosphorylation. PMID- 3040272 TI - Immunocytochemical demonstration of cyclic AMP-dependent protein kinases in duct cells of the rat parotid gland. AB - The cyclic AMP-dependent protein kinases were immunolocalized in the rat parotid gland using a monospecific antiserum against their catalytic subunit. The kinases were found to be primarily located in the cytoplasm of the parotid duct cells with a preference for the apical cell region. The result questions the traditional view of the control of parotid gland secretion and suggests a role of cyclic AMP not only in the acinar protein secretion but also in ductal functions like fluid and electrolyte transport. PMID- 3040273 TI - Development of diel rhythm of CAMP in the eye of the trout postembryo. PMID- 3040274 TI - Mechanism of polymorphonuclear leukocyte activation by myristate. Involvement of calcium ion and protein kinase C. AB - The stimulative effects of myristate on the superoxide generation and depolarization of membrane potential of polymorphonuclear leukocytes (PMN) are particularly strong, yet myristate does not affect the intracellular free Ca2+ level ([Ca2+]i) in the presence of 1 microM free calcium in calcium-EGTA buffer. The half maximum concentration of myristate was 10 microM. Myristate inhibited the transitory changes in [Ca2+]i induced by formylmethionyl-leucyl-phenylalanine (FMLP), but stimulated further the FMLP-induced superoxide generation; these effects are similar to those of phorbol myristate acetate (PMA). The myristate induced superoxide generation was partially inhibited by H-7, a specific inhibitor of protein kinase C. Myristate stimulated the activity of Ca2+- and phospholipid-dependent protein kinase (protein kinase C) in a concentration dependent manner in the presence of 10(-6) M Ca2+. The Ka was 100 microM. These results suggested that there is no relation between the superoxide generation and the [Ca2+]i change in PMNs and that the effects of myristate are similar to those of PMA against PMN. PMID- 3040275 TI - Interactions of the antitumor drug, etoposide, with reduced thiols in vitro and in vivo. AB - The interaction of activated etoposide, 4'-demethylepipodophyllotoxin-9-(4,6-O ethylidene-beta-D-glucopyra noside) (VP-16), with thiols has been studied both in vitro and in vivo in mice. We have found that both glutathione (GSH) and cysteine rapidly reduce the VP-16 free radical, which results in the regeneration of the parent drug and the oxidation of the thiol. Using spin-trapping and electron spin resonance (ESR) techniques, we have shown that this one-electron/hydrogen donation by thiols forms thiyl radicals (RS.) which are intermediates for the formation of the oxidized thiols. The administration of VP-16 in vivo to mice decreased the total thiol levels in liver and concomitantly increased the formation of oxidized thiols. Furthermore, VP-16 stimulated glutathione reductase in liver. While administration of VP-16 also increased the total thiol pools in kidney, in contrast, no significant effects were observed on lung and heart thiol pools. PMID- 3040276 TI - A circular dichroism study of the binding of CC-1065 to B and Z form poly(dl 5BrdC).poly(dl-5BrdC). AB - CC-1065, Benzo[1,2-b:4,3-b']dipyrrole-3(2H)-carboxamide, 7-[[1,6-dihydro-4 hydroxy-5-methoxy-7-[(4,5,8,8a-tetrahydro-7-methyl-4- oxocyclopropa[c]pyrrolo[3,2 e]indol-2(1H)-yl)carbonyl]benzo [1,2-b:4,3-b']dipyrrol-3(2H)-yl]carbonyl]-1,6 dihydro-4-hydroxy- 5-methoxy-, (7bR,8aS), binds to the B form of poly(dl 5BrdC).poly(dl-5BrdC) to yield a reversibly bound species whose stability with respect to an irreversibly bound species (presumably the inosine N-3 adduct) is much greater than it is for other DNA polymers. Competitive binding experiments with netropsin, show that this reversibly bound species of CC-1065 contains CC 1065 in the minor groove of the double helix. A review of the CC-1065 binding data obtained on other synthetic DNA polymers suggests that the widely different rates of species conversion shown by these polymers may result from small differences in DNA secondary structure rather than from different alkylating abilities of the adenine or inosine N-3 active site. CC-1065 converts the Z-form of poly(dl-5BrdC).poly(dl-5BrdC) in 3.5 M sodium chloride to the B form and does not bind to the Z form in this solvent system. CC-1065 bound to the B form polymer inhibits the formation of the Z form if the helix is saturated with CC 1065. Regions of the polymer without bound CC-1065 can convert to the Z form with added salt, producing a situation where the polymer contains both the B and Z conformations. In 4.0 M sodium chloride, where the Z conformation is also predominate, the addition of CC-1065 causes chiral aggregates to form, and CC 1065 binds to the aggregates. The addition of dimethylformamide in the absence of CC-1065 or a simple dilution of the 4.0 M sodium chloride polymer solution with water also causes aggregation, indicating that the Z form of this polymer in 4.0 M sodium chloride is unstable with respect to an aggregated form. PMID- 3040277 TI - [Distribution and diagnostic role of GFAP and S-100 protein in human brain tumors]. PMID- 3040278 TI - [Immunohistochemical assay of glial fibrillary acidic protein in a cultured cell line from human malignant glioma and its implanted tumor in nude mice]. PMID- 3040279 TI - [New advances in the morphological study of neuro-tumors]. PMID- 3040280 TI - [Assay of beta-adrenergic receptor function and evaluation of the preventive effects of wen yang pills in asthmatics]. PMID- 3040281 TI - [Tumors of the liver secondary to androgen therapy. Apropos of 2 cases in children]. AB - The authors report 2 cases of hepatocellular tumour in children treated with anabolic androgens for aplastic anemia. In both cases, the presentation was by a picture of acute abdomen due to hemoperitoneum caused by tumour rupture. In the first case, there was multiple hepatic adenomas necessitating right hepatic lobectomy. The second infant had a single tumour of segment IV treated by simple excision of the tumour. It was a hepatocellular-carcinoma. Follow-up for one year after the initial operation showed no signs of recurrence in both infants. The review of the literature permitted us to find 48 other cases of hepatocellular tumour secondary to androgen therapy. In order of frequency, the hepatocellular carcinoma is the most frequent and it is usually single; followed by the adenoma which is usually multiple. The other types of tumours are rare: focal nodular hyperplasia, angiosarcoma and cholangiocarcinoma. The hepatocellular-carcinoma and adenoma have some characteristic features: spontaneous regression may occur after withdrawing of androgens; the risk of rupture is important; their evolution is almost always favorable despite of a severe histopathological picture; the alpha-foeto-protein is nearly always negative; and the metastasis are exceptional. The hepatocellular-carcinomas associated with androgen therapy are probably just adenomas with marked dysplasia, but their long term malignant potential remain unknown. Except in case of rupture, surgical intervention should be postponed until the effect of discontinuing the hormonal therapy is assessed, because of the potential for spontaneous regression. The administration of antineoplastic chemotherapeutic agents should be reserved for the tumours showing evidence of malignancy. PMID- 3040282 TI - Granulocyte transfusions in neutropenic patients. AB - Patients with severe neutropenia are at increased risk for systemic infection with bacteria or fungi. This risk is in proportion to both the degree and duration of the neutropenic process. Although granulocyte transfusion as a means of augmenting host defenses would appear to be a logical therapeutic intervention in clinical contexts involving severe and prolonged neutropenia, several features of granulocyte physiology and collection complicate such considerations. These include the large numbers of granulocytes normally produced by healthy hosts, the short survival of the granulocyte in the circulation after transfusion, the relatively small number of granulocytes which can be collected using currently available pheresis techniques, problems associated with alloimmunization, and the possibility of transferring disease (CMV, toxoplasmosis, hepatitis) by means of these transfusions. In the mid-1970s, well-designed clinical studies strongly suggested that patients with documented Gram-negative sepsis or tissue infection that failed to respond to appropriate antibiotics were significantly benefited by granulocyte transfusions. With recent advances in potent, broad-spectrum antibiotic availability, some have questioned whether these observations remain valid. Several studies regarding the prophylactic use of granulocyte transfusions in patients undergoing allogeneic bone marrow transplantation and/or induction therapy for leukemia have failed to reveal therapeutic benefits and suggested the possibility of significant side effects. These studies are reviewed. PMID- 3040283 TI - [Utilization of an ELISA technic for the quantification of antipoliovirus antibodies in human sera]. AB - A double antibody enzyme linked immunosorbent assay (ELISA) was elaborated for detection of poliovirus antibodies in human sera. The IgG to be titrated were immunoabsorbed by capture on the solid phase. The antigens used were obtained from vero cell cultures (green Monkey Kidney Cells). The reaction was followed by adding rabbit antipoliovirus serum, then sheep Fab fragment prepared against rabbit IgG and labelled with horse radish peroxidase. Ortho-tolidine was used as the chromogen substrate to reveal the reaction. This enabled a first reading with the naked eye. This technique allows to keep a better track of the poliomyelitis immunization. PMID- 3040285 TI - Solid malignant tumors in children: an experience with 506 cases. PMID- 3040284 TI - [Orbitopalpebral metastasis of breast cancer]. PMID- 3040286 TI - Elevation of plasma levels of fluorinated pyrimidines by guanosine 5' monophosphate. AB - The plasma concentration of 5-fluorouracil (FUra) following the i.v. administration of FUra and guanosine 5'-monophosphate (GMP) or guanosine 5' triphosphate (GTP) was markedly elevated. These values were more than 5-fold higher than those obtained with FUra alone over 60 min after administration. The elevation of plasma levels corresponded to the dose of GMP. Higher levels of FUra were maintained in the plasma after injection of inosine or inosine 5' monophosphate in combination with FUra than after FUra alone, but they were lower than those induced by GMP or GTP. Moreover, plasma levels of two other fluorinated pyrimidines, 5'-deoxy-5-fluorouridine (DFUR) and 5-fluoro-2' deoxycytidine (FdCyd), were also elevated by GMP. The combination of DFUR and GMP resulted in higher plasma levels of DFUR itself and FUra (12- and 10-fold, respectively, 30 min after treatment). After administration of FdCyd plus GMP, the plasma levels of FdCyd, 5-fluoro-2'-deoxyuridine, which is converted from FdCyd by cytidine deaminase, and FUra were 2-, 6-, and 7-fold higher, respectively, than those after FdCyd alone 30 min after treatment. Thus, GMP is the most effective compound for the maintenance of high plasma levels of fluorinated pyrimidines. PMID- 3040288 TI - Etoposide combination therapy for small cell carcinoma of the lung. AB - Sixty-three consecutive patients with small cell carcinoma of the lung were treated by six cycles at 3-week intervals of etoposide 120 mg/m2 i.v. on day 1 and orally on days 2-5, adriamycin 40 mg/m2 i.v. on day 1 and vincristine 1.4 mg/m2 i.v. on day 1. Tumour bed irradiation was administered to patients with limited disease after chemotherapy. In limited-disease and extensive-disease patients the median survival was 12 and 6 months respectively. The 2-year survival rate (life table) in limited-disease patients was 26%. Treatment morbidity was low. A prospective randomised trial is being undertaken to further evaluate the role of oral etoposide in combination chemotherapy. PMID- 3040287 TI - Phase II clinical and pharmacological study of oral 4-demethoxydaunorubicin in advanced non-pretreated small cell lung cancer. AB - 4-Demethoxydaunorubicin (4-DMDNR) is an oral anthracycline with antitumour activity demonstrated in a number of clinical studies. We have assessed the usefulness of 4-DMDNR in 16 patients with advanced small cell lung cancer, none of whom had received previous chemotherapy. There were no complete or partial responders among the 14 evaluable patients, but 9 patients showed a minor radiographic improvement and 6 reported transient symptomatic improvement. Side effects were mostly minor or moderate, although one patient succumbed to septicaemia during neutropenia following treatment. There was no evidence of cardiotoxicity in any patient. Pharmacological studies were undertaken in 8 patients. A previously undescribed metabolite, identified as the 7-deoxyaglycone of 4-demethoxydaunorubicinol, was detected in 3 patients and these 3 patients all showed some anti-tumor response. PMID- 3040289 TI - Involvement of the prostatic steroid-binding protein in the transfer of ligand to the dioxin receptor. AB - The prostatic steroid-binding protein (PSP) represents a highly abundant protein in the rat prostate which binds carcinogens reversibly and with high affinity. The biological role of PSP and the toxicological implications of the carcinogen protein interaction are unclear. In this report, we have attempted to examine a possible role of PSP in the transfer of ligands to the dioxin receptor. PSP was purified from the rat ventral prostate and labeled in vitro with 2,3,7,8 [3H]tetrachlorodibenzo-p-dioxin (dioxin). Dioxin-labeled PSP was then incubated with rat liver cytosol in the presence or absence of a 200-fold excess of nonradioactive competitor, 2,3,7,8-tetrachlorodibenzofuran. After 2 h of incubation, a complete in vitro transfer of ligand from PSP to the rat hepatic dioxin receptor was observed, as assessed by velocity sedimentation analysis of specific dioxin binding. These results indicate that a high abundance carcinogen binding protein, such as PSP, may be of importance in the cellular transfer of dioxin receptor ligands, thereby eliciting a receptor-mediated biochemical and/or toxic response. PMID- 3040290 TI - Alterations of Na+-K+-ATPase, Ca2+-ATPase, and Mg2+-ATPase activities in erythrocyte, muscle, and liver of traumatic and septic patients. AB - Na+-K+-ATPase, Ca2+-ATPase and Mg2+-ATPase activities of erythrocyte membrane, microsomal fractions of rectus muscle, and liver were measured colorimetrically in the biopsy specimens of 14 control, 7 uncomplicated trauma (group 2), and 14 severe trauma or septic patients (groups 3-A and 3-B). In erythrocytes, these three ATPase activities in group 2 were not significantly changed but sepsis of both the acute (group 3-A) and ongoing type (group 3-B) decreased all of the ATPase activities. In muscle, there was a significant loss of three ATPase activities in the acute insult of severe trauma or sepsis (group 3-A), while Na+ K+-ATPase and Mg2+-ATPase activities were not significantly changed in ongoing, severe trauma (group 3-B). In the liver, a tendency for all three ATPase activities to decrease is noted in the severe traumatic group. However, a statistical difference between the control and severe traumatic group showed only for Na+-K+ ATPase and Mg2+-ATPase in group 3-A and Ca2+-ATPase in group 3-B. Correlation coefficients between erythrocyte, muscle, and liver for three ATPase activities are between 0.4 and 0.5. The mechanism which alters ATPase activity remains unknown in this study, but it may account for the variation in traumatic insult, in hemodynamic and hormone changes, and in tissue energy stores. PMID- 3040291 TI - Receptors for mouse erythrocytes on human immature B lymphocytes: still a valuable tool in immunohaematology. PMID- 3040292 TI - Renal vascular adjustments to partial renal venous obstruction in dog kidney. AB - Blood flow studies were conducted in neurolept anesthetized dogs to characterize the involvement of renal nerves in ipsilateral renal vasoconstriction seen during acute elevation of renal venous pressure above 30 mm Hg. Renal blood flow was measured electromagnetically. The vasoconstrictor response was almost abolished by acute surgical denervation of the kidney, since renal vascular conductance remained unchanged during renal venous pressure elevation from 30-60 mm Hg. However, following additional alpha-adrenoceptor blockade or chronic renal denervation, renal vascular conductance increased progressively during renal venous pressure elevation to 60 mm Hg. The effect of acute decapsulation of kidney was studied in another group of dogs. Decapsulation induced a vasoconstriction. The decrease in renal vascular conductance observed during renal venous pressure elevation was unaffected by acute surgical denervation of decapsulated kidney, but was almost abolished following additional alpha adrenoceptor blockade or chronic denervation. In decapsulated chronically denervated kidney, the increase in renal vascular conductance during renal venous pressure elevation to 60 mm Hg was still present but considerably attenuated as compared with the chronically denervated kidney with intact capsule. The renin angiotensin system did not participate in acute vascular adjustments to renal venous stasis in intact kidney or in decapsulated acutely surgically denervated kidney. The data favor the view that neurogenic and myogenic mechanisms significantly influence the vasoconstrictor response to renal venous pressure elevation in dog kidney. The neurogenic contribution to the vasoconstrictor response comprises intrarenal and extrarenal vasoconstrictor mechanisms evoked reflexively by renal venous pressure elevation, and the myogenic contribution to the vasoconstrictor response comprises opposing vasodilator mechanisms due to increase in renal interstitial tissue pressure during renal venous pressure elevation. PMID- 3040293 TI - Hypoxia does not alter angiotensin converting enzyme activity in hamster pulmonary microvessels. AB - Studies were initiated to investigate the effects of hypoxia on the conversion of angiotensin I (AI) to angiotensin II (AII) in microvessels of the lung. Using the technique of allografting neonatal lung tissue into the cheek pouch of normal hamsters, the microvessels of the lung, pulmonary arterioles, and venules could be visualized and manipulated by direct in vivo microscopy. The microvessels of the lung were studied 7-10 days after allografting by anesthetizing the hamster with pentobarbital (6.0 mg/100 g body weight i.p.) and then preparing the lung tissue for observation. The tissue was suffused with a Ringer's bicarbonate solution bubbled with a normal (20% O2-5% CO2-75% N2) or a low (95% N2-5% CO2) oxygen mixture. After equilibration, a pulmonary arteriole or venule was selected for observation, and the vessel geometry was recorded. Then, a micropipette containing either AI or AII was positioned alongside the vessel, and the agent was delivered continuously for 2 minutes. Lumen diameter was recorded continually for 8-10 minutes. This procedure was repeated until both angiotensins were tested on pulmonary arterioles and venules under conditions of a normal and low oxygen environment. This protocol was repeated on cheek pouch microvessels that did not contain pulmonary allografts. Both AI and AII produced rapid decreases in the lumen diameters of all microvessels tested. This vasoconstriction was greater for AII, and the oxygen environment did not alter the response. Conversion of AI to AII was not altered by the oxygen environment, and the relative conversion was similar in the microvessels of the lung and cheek pouch.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3040294 TI - Platelet-activating factor effects on bovine pulmonary artery endothelial cells. AB - Endothelial cells (ECs) were isolated from bovine pulmonary artery and maintained in long-term culture. On reaching confluency, ECs formed a characteristic "cobblestone" monolayer. One hour after addition of 1 nM platelet-activating factor (PAF) to the growth medium, ECs underwent dramatic changes in shape from their normal polygonal morphology to more elongated spindle-shaped forms. More pronounced effects were evident in the presence of 0.1 nM phorbol-12-myristate-13 acetate (PMA), a potent activator of C kinase. It was found that at concentrations from 10(-11)-10(-7) M, PAF stimulates the phosphoinositide turnover in EC. The half-maximal activation in the release of inositol phosphates was at 10(-9) M. This finding suggested that an increase in intracellular Ca2+ concentration and activation of protein kinase C were involved in the mechanism of action of PAF on EC. The metabolic responses of EC were evaluated by measuring the activity of beta-adrenergic receptor-coupled adenylate cyclase (AC) in a crude membrane fraction and by assay of prostacyclin and thromboxane released by cultured EC. AC from control membranes was activated by isoproterenol in a dose dependent manner (EC50 = 30 nM) from 0.8-5.5 pmol cAMP/min/mg protein. If the membranes were isolated after preincubation of ECs with 1 nM PAF or 0.1 nM PMA, the AC activity was decreased by 70 and 90%, respectively; in both cases, affinity for isoproterenol was lowered threefold (EC50 = 100 nM). Our data suggest that PAF interaction with EC leads to an apparent beta-adrenergic receptor desensitization that probably acts via a phosphorylation mechanism involving C kinase.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3040295 TI - Pertussis toxin-insensitive phosphoinositide hydrolysis, membrane depolarization, and positive inotropic effect of carbachol in chick atria. AB - Muscarinic agonists can stimulate rather than inhibit cardiac muscle in some preparations. In left atria from hatched chicks, treatment with pertussis toxin reversed the membrane action of carbachol from hyperpolarization to depolarization and reversed the inotropic effect of carbachol from negative to positive. Acetylcholine also depolarized the membrane and increased the force of contraction in atria from pertussis-toxin-treated chicks although oxotremorine did not. These cholinergic responses were blocked by atropine but not by adrenoceptor antagonists, suggesting that they are mediated via muscarinic receptors and are not due to actions of endogenously released catecholamines. Muscarinic receptor stimulation leads to two distinct biochemical responses in chick atria: inhibition of adenylate cyclase and activation of phosphoinositide (PI) hydrolysis. The former is lost in atria from pertussis-toxin-treated chicks, whereas the PI response persists. The pharmacologic characteristics of the PI response resemble those of the depolarization and positive inotropic response. Both are insensitive to blockade by pertussis toxin, require high concentrations of carbachol, and are elicited by acetylcholine but not by oxotremorine. The present study suggests that muscarinic agonist-induced PI turnover may be responsible for the membrane depolarization and positive inotropic effects of carbachol and acetylcholine; that an increase in Na+ conductance underlies these responses; and that it is stimulated either by an increase of intracellular calcium mobilized by inositol triphosphate and/or by activation by protein kinase C. PMID- 3040296 TI - Mechanisms of flunarizine-induced vasodilation in the rabbit mesenteric artery. AB - The vasodilating effects of flunarizine on smooth muscle strips of rabbit mesenteric artery have been investigated and compared with those of nifedipine. Flunarizine (30-300 nM) dose-dependently inhibited Ca2+-induced contractions in Ca2+-free solution containing 100 mM K+. Double reciprocal analysis showed that this inhibition was either competitive at low concentrations (30-100 nM; nifedipine-like) or noncompetitive at high concentrations (0.3-1 microM). The latter seemed to be partly related to an inhibition of contractile proteins as estimated from Ca2+-induced contractions in saponin-treated chemically skinned muscle strips. In contrast to the actions of nifedipine, flunarizine inhibited norepinephrine (NE)-induced contractions more than those induced by high K+, and at 0.3 microM, this agent totally blocked NE-induced contraction. Flunarizine also inhibited NE-induced contraction in Ca2+-free solution containing 2 mM EGTA. In Ca2+-free solution, NE rapidly hydrolyzed phosphatidylinositol 4,5 bisphosphate (PI-P2) and produced phosphatidic acid (PA). Flunarizine (30 and 300 nM), but not nifedipine (100 nM), inhibited NE-induced hydrolysis of PI-P2 and production of PA. However, flunarizine (100 nM) did not modify the contraction induced by 10 microM inositol 1,4,5-trisphosphate in chemically skinned muscle strips. It is concluded that flunarizine inhibits both voltage-dependent (nifedipine-like) and receptor-operated Ca2+ influx induced by NE and also inhibits NE-induced Ca2+ release from intracellular stores due to inhibition of the hydrolysis of PI-P2. PMID- 3040297 TI - Vitamin D-binding protein synthesized by a human hepatocellular carcinoma cell line. AB - The binding protein for 25-hydroxycholecalciferol was studied in the medium spent for the culture of HuH-7 cells, which were originally derived from a human hepatocellular carcinoma tissue. The binding protein for 25 hydroxycholecalciferol synthesized by HuH-7 cells was immunologically similar to vitamin D-binding protein in human serum and had an inter-alpha mobility. A sedimentation coefficient of 4.1 S was found on sucrose density gradient analyses. The molecular weight was estimated to be approximately 58,000 by gel filtration on a standardized column of Sephadex G-150. When mixed with filamentous actin purified from rabbit skeletal muscle, it depolymerized filamentous actin and bound to monomeric, globular actin to make a 5.5 S 1:1 molar complex with a molecular weight of approximately 100,000. These results support the conclusion that HuH-7 cells produce a functional vitamin D-binding protein. PMID- 3040298 TI - Structurally abnormal transthyretin causing familial amyloidotic polyneuropathy in Sweden. PMID- 3040299 TI - Pattern of plasma levels of cortisol, dehydroepiandrosterone and pregnenolone sulphate in normal subjects and in patients with homozygous familial hypercholesterolaemia during ACTH infusion. AB - The aim of this study was to determine whether patients with homozygous familial hypercholesterolaemia (FH) have impaired adrenal cortical function. Plasma levels of cortisol, dehydroepiandrosterone (DHA), pregnenolone sulphate (PS) and DHA sulphate (DHAS) were measured during and 8 h ACTH infusion in six controls and two patients with homozygous FH. The basal PS levels of both patients and the basal DHA level of one were abnormally low for age and pubertal stage. During ACTH infusion we observed in both patients: (1) a mild impairment of control response after sustained stimulation (P less than 0.002); (2) a clear impairment of PS response (values less than 2 SD of those in controls); (3) a clear impairment of DHA response (values less than 2 SD) until 4 h in the boy who was at pubertal stage 4 and whose response could be compared to controls; no increase at all in the affected girl (pubertal stage 2) at a stage where normal subjects respond with significant increase. These results suggest that patients with FH lack cholesterol for corticosteroid biosynthesis under maximal ACTH stimulation and that mild chronic ACTH stimulation due to a deficit in the cholesterol supply to adrenal cells might increase the conversion of delta 5 to delta 4-steroids. They provide further evidence to support the primordial role of low density lipoprotein (LDL)-cholesterol in adrenal steroidogenesis in vivo. PMID- 3040300 TI - Analysis of the relation between alopecia and resistance to 1,25-dihydroxyvitamin D. AB - Alopecia is a frequent feature in hereditary resistance to (1,25(OH)2D). We sought insight into this feature by analysing data from affected members of 30 kindreds. We assessed indices of mineral metabolism in one group with normal hair compared with a group with alopecia. Hereditary resistance to 1,25(OH)2D was diagnosed at an earlier age in alopecic patients (0.9 vs 3.3 years, P less than 0.05); this reflected late presentation of metabolic bone disease in some cases with normal hair and could not be attributed to early diagnosis resulting from the striking feature of alopecia. For untreated subjects, serum concentrations of calcium and 1,25(OH)2D were similar in both groups of patients. During calciferol therapy, however, the cases with alopecia showed lower serum calcium (1.9 vs 2.4 mmol/l, P less than 0.005), but higher serum 1,25(OH)2D (2900 v 340 pg/ml, P less than 0.005). Hair status did not predict the type of defect identified with cultured skin fibroblasts but did correlate with responsiveness of 25(OH)D 24 hydroxylase to 1,25(OH)2D3 in those cells. Cells from seven of eight kindreds with alopecia showed no 24-hydroxylase response to high doses of 1,25(OH)2D3 while cells from five of six kindreds with normal hair showed a 24-hydroxylase response to high doses of 1,25(OH)2D3. We conclude that in cases with hereditary resistance to 1,25(OH)2D alopecia reflects the more severe grades of this resistance based upon earlier age at time of diagnosis, lower potential for calcaemic response to calciferols, and lower potential for 24-hydroxylase response to 1,25(OH)2D3 by cultured skin fibroblasts. PMID- 3040301 TI - An analogue of met-enkephalin attenuates the pituitary-adrenal response to ovine corticotrophin releasing factor. AB - The met-enkephalin analogue, DAMME, suppresses the pituitary-adrenal axis in normal subjects; it is not clear whether this occurs at the level of the pituitary or above. We therefore administered synthetic ovine corticotrophin releasing factor (CRF-41) 100 micrograms i.v. to a group of normal male subjects, with or without pretreatment with DAMME 250 micrograms i.v., and monitored the response of plasma ACTH and serum cortisol. CRF-41 caused a marked stimulation of ACTH and cortisol release, but this was significantly attenuated by pretreatment with DAMME. It is therefore concluded that DAMME either directly inhibits the corticotroph at the level of the pituitary, or that it suppresses release of an additional factor which normally potentiates the action of CRF-41 on the pituitary. PMID- 3040302 TI - Increased atrial natriuretic factor receptor density in cultured vascular smooth muscle cells of the spontaneously hypertensive rat. AB - To explore the role of the atrial natriuretic factor (ANF) system in the pathophysiology of hypertension we examined the binding kinetics of synthetic ANF to cultured vascular smooth muscle cells (VSMCs) derived from the spontaneously hypertensive rat (SHR) and two normotensive controls-the Wistar Kyoto (WKY) and American Wistar (W). The number of maximal binding sites (Bmax) per cell (mean +/ SEM; X10(3] were: SHR = 278.0 +/- 33.0, WKY = 28.3 +/- 7.1 and W = 26.6 +/- 4.2. The differences between the SHR and normotensive strains were significant at p less than 0.001. The equilibrium dissociation constant (Kd; X 10(-9)M) was higher in SHR VSMCs (0.94 +/- 0.14) than in WKY (0.22 +/- 0.09; p less than 0.01) and W (0.39 +/- 0.14; p less than 0.02) cells. The plasma levels of the immunoreactive ANF were higher in SHR than the normotensive controls. We suggest that the relatively greater ANF receptor density in cultured VSMCs of the SHR represents a response to the in vitro environment which is relatively more deficient in ANF for VSMCs of the SHR as compared with the normotensive rats. Thus, the capacity of the SHR VSMC to regulate ANF receptor density appears to be independent of the blood pressure level. PMID- 3040303 TI - Decreased Na+K+ATPase activity in the aortic endothelium and smooth muscle of the spontaneously hypertensive rats. AB - The activity of Na+K+ ATPase in the endothelium and smooth muscle of the aortae of normotensive and hypertensive rats was investigated. The enzyme activity in the endothelium and smooth muscle of the spontaneously hypertensive rats (SHR) was 2.15 +/- 0.48 and 12.98 +/- 0.99 respectively. These values were significantly lower (P less than 0.05) than the enzyme activity in the corresponding tissues (10.10 +/- 1.78 for endothelium, 20.77 +/- 2.54 for smooth muscle) of the normotensive Wistar Kyoto (WKY) rats. However, with the low blood pressure spontaneously hypertensive rats (LBP-SHR) i.e. in those animals whose blood pressures were below 150 mm Hg, the enzyme activity in both tissues was not significantly different from those of the WKY. Since Na+ K+ ATPase is coupled to the sodium-potassium pump whose activity affects the functions of other pumps, the results indicate that the development of high blood pressure in the SHR may be related to an alteration in the transport of cations across the cell membrane. PMID- 3040305 TI - Some properties of erythrocyte Na+-K+-ATPase in essential hypertension. AB - The activity and some allosteric properties of Na+-K+-ATPase in erythrocytes and their membrane preparations (ghosts) from 57 patients with essential hypertension and 36 normotensive controls were studied. To reveal enzyme activity in whole erythrocytes the cells were pretreated with detergent Tween-20. It was found that in the patient erythrocytes the Na+-K+-ATPase activity was 33% less as compared to the control group. Moreover, in the patient erythrocytes the sensitivity of the enzyme to high concentrations of MgCl2 was decreased. In contrast, no analogous changes of the enzyme were revealed in the patient ghosts. It is suggested that the erythrocytes of patients with essential hypertension contain an inhibitor of Na+-K+-ATPase. PMID- 3040304 TI - Assay of a circulating sodium pump inhibitor in patients with essential hypertension and normotensive subjects. AB - The plasma levels of a sodium pump inhibitor (Na+ PI) were measured by a modified method of Hamlyn et al, using dog kidney Na+, K+-ATPase. When the level of Na+ PI was expressed as the % inhibition of the enzyme and compared with that of a control solution, it was found to be 9.0 +/- 0.7% in 43 untreated patients with essential hypertension. This was significantly higher than 5.0 +/- 0.4% for 56 normotensive subjects (p less than 0.01). Male patients with essential hypertension showed the highest mean value of 10.5 +/- 1.1%, disclosing an apparent sex difference in the patient group (p less than 0.01). Only in female patients was there a significant positive correlation between the inhibitor's level and the mean blood pressure (r = 0.649, p less than 0.01). These results provided additional evidence for increased Na+ PI in the plasma of patients with essential hypertension, which might bear an important role in the pathogenesis of the disease. PMID- 3040306 TI - Ouabain-like and non-ouabain-like factors in plasma of patients with essential hypertension. AB - Circulating inhibitor of Na+,K+-ATPase and ouabain-like immunoreactivity were studied in patients with essential hypertension. In the plasma of patients, two types of Na+,K+-ATPase inhibitors (ouabain-like and non-ouabain-like inhibitors) and ouabain-like immunoreactivity were detected. Ouabain-like inhibitor was clearly detected at a low KCl concentration (0.1 mM) in the assay buffer, and non ouabain-like inhibitor was detected at a high KCl concentration (10 mM). The plasma level of ouabain-like inhibitor correlated significantly with that of ouabain-like immunoreactivity (p less than 0.001) and with a mean blood pressure (p less than 0.01). The plasma level of non-ouabain-like inhibitor was not correlated with the levels of either ouabain-like immunoreactivity or mean blood pressure. The level of plasma ouabain-like inhibitor did not correlate with that of plasma non-ouabain like inhibitor. Both ouabain-like inhibitor and ouabain like immunoreactivity in the plasma of patients with essential hypertension were significantly higher than those in normotensive subjects, but the plasma level of non-ouabain-like inhibitor in patients with essential hypertension was not higher than that in normotensive subjects. These results suggest that the plasma from patients with essential hypertension contains ouabain-like factor(s) which is important to maintain the high blood pressure. PMID- 3040307 TI - A comparative study on the release of leukotrienes and histamine by guinea pig lung and trachea after challenge with antigen or stimulation with ionophore A23187 or melittin. AB - The release of leukotrienes and histamine from guinea pig lung and trachea after immunological and nonimmunological stimulation were compared. Antigen, ionophore A23187 and melittin caused the release of leukotriene (LT)B4, LTC4, LTD4 and LTE4 from lung and trachea as determined by reverse-phase high performance liquid chromatography (RP-HPLC) and bioassay. The release of LTB4 by lung and trachea was maximum after 5 min of ionophore stimulation (128 +/- 40 and 142 +/- 29 pmol/g tissue, respectively). Lung, but not trachea, also released the 20-OH-LTB4 and 20-COOH-LTB4. The release of LTC4 by lung tissues was maximum after 5 min, whereas maximal tracheal responses occurred at 10 min (27 +/- 11 and 9 +/- 3.5 pmol/g tissue, respectively). Maximal release of LTD4 by lung and trachea respectively occurred after 10 and 15 min (103 +/- 21 and 20 +/- 6 pmol/g tissue, respectively). The release of LTD4 in response to ionophore by both tissues decreased after 15 min, whereas the release of LTE4 continued to increase. Release of leukotrienes from melittin stimulated lung was 2-3-fold less than in ionophore stimulation. In contrast, tracheal responses to melittin and ionophore for the release of LTB4 were equivalent, whereas release of peptidoleukotrienes in response to melittin was approximately 50% that resulting from ionophore. Antigen challenge was the least potent stimulus for LTB4 release in both tissues, whereas it was at least as potent as melittin for the release of peptidoleukotrienes. The release of histamine by lung tissue was approximately 2 3-fold greater than by trachea (7 +/- 1 and 2 +/- 0.5 nmol/g tissue, respectively) after 5 min of stimulation with either ionophore, melittin or antigen. These data demonstrate that lung tissues and trachea respond to immunologic stimulations by releasing the mediators of inflammation and immediate hypersensitivity. The lung releases peptidoleukotrienes and histamine 2-5-fold greater than the trachea, whereas the release of LTB4 in both tissues are approximately equal. PMID- 3040308 TI - A single cell assay for the study of gamma-interferon formation in leprosy patients. AB - The number of gamma-interferon producing cells in the peripheral blood of leprosy (LL and BT) patients and controls was studied by the reversed protein A plaque assay before and after exposure in vitro to Mycobacterium leprae bacilli and Epstein-Barr virus (EBV). The level of spontaneous gamma-interferon production was significantly higher in BT patients compared to LL patients and controls. Mycobacterium leprae induced a specific gamma-interferon response in lymphocytes from BT patients and from healthy contacts whereas in LL patients and non-exposed controls the response was low or non-existing. There were no significant differences in the gamma-interferon response to EBV between the above groups. PMID- 3040310 TI - Role of endogenous prostaglandins in gastric secretion and mucosal defense. AB - Prostaglandins are found in high concentration in the gastric mucosa and gastric juice. Exogenous prostaglandins inhibit acid secretion, stimulate mucus and bicarbonate secretion, alter mucosal blood flow, and provide dramatic protection against a wide variety of agents which cause acute mucosal damage. The physiological role of prostaglandins is still being elucidated. There is now strong evidence that endogenous prostaglandins modulate acid secretion by blocking the histamine-stimulated increase in cyclic AMP within the parietal cell. This function is probably controlled by intraluminal pH. It is likely that mucus and bicarbonate secretion by both stomach and duodenum are influenced by endogenous prostaglandins. A physiological role of prostaglandins in mucosal protection is less certain. Prostaglandins are released by trivial injury, and this probably serves a defensive function. A mucosa which is prostaglandin depleted is more susceptible to damage, but does not spontaneously ulcerate. It is conceivable that peptic ulcer disease may be in part caused by an impaired mucosal prostaglandin response to food. PMID- 3040309 TI - Secondary immune amplification following live poliovirus immunization in humans. AB - Eight subjects inoculated orally with live attenuated poliovirus were investigated to study the effects of live virus infection on human T-cell responses. Proliferation to poliovirus and unrelated recall antigens were measured serially over a 3-week period. Five of eight subjects inoculated demonstrated a clear anamnestic response to poliovirus, but three did not. Only the five subjects demonstrating an anamnestic response to poliovirus were found to have augmented secondary immune responses to two unrelated recall antigens (tetanus toxoid and reovirus) and in the autologous mixed lymphocyte response (AMLR). No consistent changes were found in circulating T-cell surface activation antigens whether or not the subjects responded to poliovirus. These studies suggest that an asymptomatic poliovirus infection associated with immunization in humans can induce nonspecific secondary immune amplification as measured by in vitro T-cell proliferative response. This amplification pathway is a potential mechanism for immune responses against antigens other than those of the infecting virus. PMID- 3040311 TI - Mineral uptake by the femora of older female X-linked hypophosphatemic (HYP) mice but not older male HYP mice. AB - X-linked hypophosphatemic (Hyp) mice are a model of human sex-linked vitamin D resistant rickets. Young adult Hyp mice are characterized by osteomalacia and decreased bone mineral content. However, older heterozygous Hyp female mice increase in bone mineral content with age so that by one year of age the bone mass/mm femoral length equals or exceeds normal females. To test for the occurrence of this mineral accretion in Hyp male mice and in homozygous Hyp female mice, femora from all 3 Hyp genotypes as well as normal male and female mice were analyzed at various ages from one to 52 weeks of age. Compared to normal mice, all three Hyp genotypes were depressed in bone ash, femoral length, and ash/length ratio at 13 weeks of age. After that age the femora of both heterozygous and homozygous Hyp females showed a slow mineral accretion and, by 52 weeks of age, a normal ash/length ratio. However, the femora of Hyp males, as well as those of normal males, failed to increase in bone mineral content or ash/length ratio after 13 weeks of age. The differences between male and female Hyp mice could not be explained by differences in the plasma levels of calcium, phosphate, or alkaline phosphatase. Increased bone mineral content in older Hyp mice was seen in both heterozygous and homozygous females but not in hemizygous males. Thus, the basis for this increase is not incomplete dominance of the Hyp gene in females nor the Lyon hypothesis. The accretion of mineral in older female Hyp mice despite lifelong reduced plasma phosphate levels suggests that there are factors other than phosphate that also regulate mineral accretion in this bone disease. PMID- 3040313 TI - [Sibling cases of membraneous lipodystrophy (Nasu) associated with neuropathy]. PMID- 3040312 TI - [Polyneuropathy in carbamate (m-tolyl methylcarbamate) poisoning]. PMID- 3040314 TI - Congenital cytomegalovirus infection with osteolytic lesions: use of DNA hybridization in diagnosis. AB - A case of congenital cytomegalovirus (CMV) infection with long-bone lesions is presented. The bone lesions consisted of broad regions of generalized osteopenia with irregular fragmentation and spiculization at the zone of provisional calcification. Diagnosis of CMV infection was made by DNA spot hybridization of the urine sediment to DNA from the Towne strain of CMV, demonstrating the usefulness of DNA hybridization for identification of CMV in clinical specimens. Bone lesions associated with congenital CMV infection are useful early clues to diagnosis but can be indistinguishable from those of congenital rubella syndrome. PMID- 3040315 TI - The balance between vascular alpha- and beta-adrenoceptors is not changed in the elderly. AB - It has been suggested that beta-adrenoceptor-mediated functions are diminished with aging and that these responses are reduced to a greater extent than are alpha-adrenoceptor-mediated responses. The resulting imbalance in the elderly may produce an increased vascular resistance from the unopposed alpha-adrenoceptor stimulation in the peripheral vasculature. To evaluate this hypothesis, we studied 12 healthy elderly and 12 healthy young subjects during a graded infusion of epinephrine and compared blood pressure response, vascular resistance, and calf blood flow as determined by venous occlusion plethysmography. In both groups, heart rate increased, blood flow to the leg increased, and vascular resistance fell in response to epinephrine infusion, but in the elderly the systolic blood pressure failed to rise as it did in the young subjects. From these data we conclude that the overall vascular response to epinephrine does not change with age and that the balance between beta-adrenoceptor-mediated vasodilation and alpha-adrenoceptor-mediated vasoconstriction is therefore unchanged in the elderly. PMID- 3040316 TI - Beta-adrenergic receptors and oxygen transport. AB - We investigated whether stimulation of baboon erythrocyte beta-adrenoreceptors affects oxygen transport by haemoglobin. To assess oxygen transport we measured the PO2 at which the haemoglobin was 50% saturated (P50) and the Hill parameter 'n'. Blood at PO2s ranging from 10 to 90 mm Hg was exposed to a 10(-3) M concentration of the agonists l-isoproterenol, l-epinephrine and l-norepinephrine in the presence and absence of 10(-5) M dl-propranolol. None of the adrenergic agents which were used in these experiments produced significant changes in either 'n' or P50 and the concentrations of 2,3-diphosphoglycerate and lactate were not altered by the agonists. We conclude from these results that short-term adrenergic stimulation of the baboon erythrocyte beta-adrenoreceptor does not affect factors known to influence oxygen transport, oxygen delivery or haemoglobin itself. PMID- 3040317 TI - Role of growth factors and their receptors in the control of normal cell proliferation and cancer. AB - Polypeptide growth factors modulate proliferation of nontransformed cells in vivo and in vitro, while cancer appears to reflect an alteration of growth-regulatory mechanisms found in normal cells. Some provocative clues for understanding the cellular biochemical events involved in growth regulation have come from the study of transforming retroviral oncogenes. Some of these oncogenes encode proteins similar to those implicated in growth factor-mediated growth control. Of particular interest is the study of growth factor receptors present on the cell surface, which are cellular homologs of members of the largest class of oncogenes, the tyrosine kinases. It is likely that the study of the interplay of growth factors, and the molecular basis of pleiotropic effects elicited by growth factors, will help to explain how growth factor-signaling pathways affect gene expression and cell division in normal and transformed states. PMID- 3040318 TI - Biochemical mechanisms of tumor invasion and metastases. AB - Tumor invasion and metastases is the major cause of treatment failure for cancer patients. There is a great need to develop new clinical methods to predict the clinical aggressiveness of a patient's tumor and to identify and eradicate clinically silent micrometastases. Such new methods may be derived from basic research into the biochemical mechanisms of invasion and metastases. We have isolated proteins involved in tumor cell attachment, invasion, and locomotion. The 'laminin receptor' is a tumor cell surface protein which specifically binds laminin, a glycoprotein of basement membranes. The laminin receptor may play a role in tumor cell attachment. 'Type IV collagenase' is a metalloproteinase which cleaves type IV basement membrane collagen but not interstitial collagens. The 'autocrine motility factor' is a secreted protein which binds to the cell surface and profoundly stimulates cell locomotion. All of these proteins appear to be augmented in actively metastatic tumor cells, at least in the models studied. They may provide strategies for diagnosis and therapy of metastases. PMID- 3040319 TI - Heparin-binding angiogenesis factors: detection by immunological methods. AB - Immunological methods for the detection of basic fibroblast growth (FGF) are described. Polyclonal antibodies directed against synthetic peptides representing amino-, internal, and carboxy-terminal regions of basic FGF, were raised in rabbits. Five techniques, enzyme-linked immunosorbent assay, immunoblot, electrophoretic transfer (Western) blot, immunoprecipitation and radioimmunoassay, were used to detect basic FGF. These techniques were used to demonstrate that a human hepatoma cell line synthesizes a growth factor structurally related to brain and pituitary basic FGF. PMID- 3040320 TI - Transduction of signals in the activation of T lymphocytes: relation to leukemia. AB - The biochemical events initiated by mitogen in T lymphocytes are the subject of this paper. Following interaction of the mitogen with its receptors, a transmembrane 'trigger-type' signal is propagated which has both positive and negative correlates. The negative signal occurs with high mitogen concentrations and is associated with membrane freezing, microtubular aggregation, receptor capping, adenylate cyclase activation, and cellular cyclic AMP increases. The positive signal occurs with optimal mitogen concentrations and is associated with changes in membrane permeability and transport with influx of calcium and potassium ion and efflux of sodium, in transport processes for glucose, amino acids, and nucleosides, and in a collected series of early membrane lipid changes which can be considered essential for the positive signal. These lipid changes include the uptake of arachidonic acid and other fatty acids, choline, phosphate and other molecules, their incorporation into membrane phospholipids, particularly phosphatidylinositol (PI), and a turnover of PI with the production of inositol triphosphate, which can be related to calcium mobilization and diacylglycerol which activates a cytoplasmic protein kinase C. A key event associated with mitogen action is arachidonic acid release. Arachidonic acid may give rise to prostaglandins and thromboxanes as part of negative components of the signal through effects on the adenylate cyclase/cyclic AMP system. Arachidonic acid gives rise to eicosanoids like 5-, 11-, possibly 12- and 15 hydroxyperoxy and hydroxy eicosatetraenoic acids and leukotrienes B4 and C4. The activation of the 5-lipoxygenase, a critical calcium-dependent step, leads via the production of 5-HPETE and 5-HETE to the activation of membrane and soluble guanylate cyclase and the production of cyclic GMP. Cyclic GMP appears to be essential for mitogen activation and is associated with cyclic GMP-dependent protein kinase activation and the phosphorylation of a number of substrates. Calcium ion influx is clearly central to mitogen action. Calcium through its influx and mobilization from cellular stores is thought to contribute directly and indirectly through the action of calmodulin and protein kinase C to the activation of a number of enzymatic processes involved in the positive signal including phospholipase C, diglyceride kinase and lipase, 5-lipoxygenase, and guanylate cyclase. Cyclic GMP and calcium ion both participate in nuclear processes leading to RNA and protein synthesis. Interleukin 2 is associated with midcycle increases in cyclic GMP and entry into DNA synthesis.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3040321 TI - A clinical comparison of the anticalculus effect of two commercially-available dentifrices. PMID- 3040322 TI - Hepatic mixed function oxidase system and enzymatic glucose metabolism in rats. AB - Therapy with enzyme inducing drugs may improve glycemic control in patients with non-insulin-dependent diabetes mellitus. We evaluated the role of a mixed function oxidase system on glucose metabolism with an animal model. Rats were treated with an inducer (phenobarbital), an inhibitor (cimetidine) and a hepatotoxin (carbon tetrachloride) for a week to cause alterations in the liver. The mixed function oxidase system was assayed by determination of the cytochrome P-450 content and NADPH cytochrome c reductase in liver. Carbohydrate metabolism was evaluated by determining blood glucose, enzymes associated with glucose phosphorylation in the liver (glucokinase, hexokinase), glucose storage as glycogen and enzymatic delivery, glucose-6-phosphatase, and peripheral tissue by determining phosphorylating enzyme (hexokinase) and a key glycolytic enzyme (pyruvate kinase) and glycogen content in muscles. The therapy with the inducer enhanced glucose utilization in liver and storage in muscles. The inhibitor decreased the mixed function oxidase system, reduced glucose phosphorylating, but not gluconeogenetic enzymes, in the liver and increased glycolysis in muscles. Carbon tetrachloride, a hepatotoxin, impaired mixed function oxidase, glucose phosphorylating and delivering enzyme activity in liver, reduced blood glucose and caused glycogen accumulation in muscles. The function of liver microsomal enzyme system seems to be closely related to enzymatic glucose metabolism in the liver and muscles. PMID- 3040323 TI - Imaging Barrett's oesophagus. AB - Eight patients who had Barrett's oesophagus confirmed by histology are presented. All had barium swallow examinations and technetium-99m sodium pertechnetate (99Tcm-pertechnetate) scintigraphy, the findings of which are reviewed and discussed. The most valuable diagnostic signs on barium swallow examination related to the site of the stricture and the presence of mucosal outpouchings. Scintigraphy was positive in all eight cases. Patients suspected of having Barrett's oesophagus on barium swallow examination should undergo scintigraphy. When the latter is positive, the endoscopist should be alerted to obtain biopsies not only at the site of stricture but serially as far as the gastrooesophageal junction. PMID- 3040324 TI - Screening of degradative enzymes from articular cartilage in experimental osteoarthritis. AB - Sixteen rabbits were killed 12 weeks after sectioning of the right knee anterior cruciate ligament. The left unoperated knee served as a control. The surface area of fibrillated cartilage from femoral condyles and tibial plateau was evaluated and expressed as a percentage of articular surfaces area. Cartilage from the femoro-patellar surfaces was homogenized for the quantification of several degradative activities, based on the release of digested products. Acid phosphatase, several glycosidases and neutral protease activity from the operated joint cartilage were significantly elevated, while collagenolytic activity was unmodified. The percentage of fibrillated cartilage correlated positively with arylsulfatase, glucosidase and neutral protease but negatively with mannosidase and fucosidase. The results may be consistent with the hypothesis of a sequential degradative process leading to cartilage destruction. PMID- 3040325 TI - Angiotensin converting enzyme (ACE) in scleroderma. PMID- 3040326 TI - The biology and treatment of warts. PMID- 3040327 TI - T and B peripheral blood lymphocytes in normal and lymphocytotic sheep. AB - Surface immunoglobulins (SIg), Peanut Agglutinin (PNA), spontaneous erythrocyte rosette (E-rosette) and Helix pomatia (HP) marker were investigated in normal and Bovine leukemia virus (BLV)-infected sheep. In normal sheep, 19.3% +/- 4.9 of peripheral blood lymphocytes (PBL) were SIg+, whereas 58% +/- 5.69 were PNA+, and 19.6 +/- 5.2 were E-rosette forming cells (E-RFC). In BLV-induced lymphocytotic sheep, SIg+ cells in PBL reached 59.4% +/- 15.06. In the same animals, PNA bound to 20.6% +/- 9.69 and E-RFC were 8.7% +/- 4.5. A panning technique was applied with an anti sheep-immunoglobulins coated plates to separate SIg+ (adherent cells = A) and SIg- cells (non-adherent cells = NA). The (A) population was 94-95% SIg+ cells and 2-3% PNA+, while the (NA) population was 0-4% SIg+ and 79-85% PNA+ cells. Thus PNA is a T cell marker in sheep species. HP, a marker for bovine T lymphocytes was also studied. Sheep PBL do not bind to HP. However, after panning separation about 50% of NA cells became HP+. PMID- 3040328 TI - Defective transport of pyrazolopyrimidine ribosides in insensitive Trypanosoma cruzi wild strains is a parasite-stage specific and reversible characteristic. AB - 1. By using freshly isolated blood trypomastigotes of twelve T. cruzi wild type strains we have found eight strains sensitive to FoB and FoA, while four and one were FoA- and FoB-insensitive respectively to the drug-mediated growth inhibition. 2. This was not so for APPR, to which most strains were transitory insensitive except two which were clearly sensitive. 3. All these pyrazolopyrimidines blocked trypomastigote-amastigote transformation. 4. Incubation of pyrazolopyrimidine-insensitive wild strains with [3H]FoA, [3H]FoB and [14C]APPR respectively indicates that insensitive cells can only accumulate low concentrations of phosphorylated metabolites. 5. This is due to a defective or impaired pyrazolopyrimidine riboside transport system in the wild type insensitive cells, as we did not detect significant variations in the levels of the various nucleoside and nucleobase metabolism enzymes studied. 6. Additional experiments suggested that FoA and FoB are incorporated by different nucleoside transport systems, as Y and ES strains were FoA-insensitive but FoB-sensitive. 7. Epimastigotes of the same T. cruzi strains were highly sensitive to low concentrations of the three pyrazolopyrimidine ribosides studied. However, when this parasitic form was allowed to transform into trypomastigotes, these cells showed the same pyrazolopyrimidine sensitivity found before, suggesting that in T. cruzi pyrazolopyrimidine riboside-insensitivity is a parasite-stage specific and reversible biochemical characteristic. PMID- 3040329 TI - The saliva of the medicinal leech Hirudo medicinalis--I. Biochemical characterization of the high molecular weight fraction. AB - 1. A method is described for obtaining dilute Hirudo medicinalis saliva by feeding leeches through a membrane on arginine/saline and squeezing them immediately after from the posterior end forwards. The process can be repeated at intervals. Yields are considerably higher than from salivary gland extracts. 2. Hirudo saliva contains hirudin, eglin, hyaluronidase, collagenase and apyrase. Leech collagenase and apyrase are here reported for the first time. 3. On gel filtration of lyophilized saliva, activity peaks were well defined. Approximate molecular weights were determined. Apyrase appears in two forms with optimum activity around pH 7.5. Collagenase was identified as belonging to the mammalian type. PMID- 3040330 TI - Enzymes with phosphoglycolate phosphatase activity in chicken skeletal muscle and liver. AB - 1. Four enzymes with phosphoglycolate phosphatase (EC 3.1.3.18) activity have been detected in extracts of chicken skeletal muscle and liver analyzed by gel filtration and ion-exchange chromatography. 2. Two enzymes have been found in muscle extracts. One of them acts on glycerate 2,3-P2, in addition to glycolate 2 P. 3. Liver extracts contain two additional enzymes with broad specificity. PMID- 3040331 TI - Renaturation of Leishmania donovani 3'-nucleotidase following sodium dodecyl sulfate-polyacrylamide gel electrophoresis. AB - 1. Renaturation of a 3'-nucleotidase from the surface membrane of Leishmania donovani promastigotes was achieved following polyacrylamide gel electrophoresis (PAGE) in the presence of sodium dodecyl sulfate (SDS). 2. Enzyme activity was detected in situ in gels, following SDS removal, by incubating the gels in reaction mixtures containing 3'-AMP or 3'-UMP as substrate followed by staining for the inorganic phosphate (Pi) reaction product with malachite green-molybic acid solution. 3. Conditions for the removal of SDS by diffusion and for the renaturation of enzyme activity are described including evidence for the detergent requirement, which is best satisfied by 3[(3-cholamidopropyl) dimethylammonio]2-hydroxy-1-propane sulfonate (CHAPSO). 4. Results indicate that the 3'-nucleotidase migrates under these conditions as a polypeptide with an Mr of 43,000. PMID- 3040332 TI - Specificity of membrane complement receptor type three (CR3) for beta-glucans. AB - The binding of the iC3b receptor (CR3) to unopsonized zymosan was shown to result from CR3 attachment to cell wall beta-glucans. A specificity of neutrophil responses for beta-glucan was first suggested by a comparison of yeast (Saccharomyces cerevisiae) cell wall components for stimulation of a neutrophil superoxide burst. Neutrophils responded poorly to heat-killed yeast, but gave increasingly better responses to cell wall polysaccharides devoid of proteins (zymosan) and nearly pure beta-glucan particles derived from zymosan. Zymosan triggered a burst that was 29% as great as that stimulated by phorbol myristate acetate (PMA), and beta-glucan particles stimulated a burst that was 72% as great as that produced by PMA. Phagocytic responses to yeast were also inhibited by soluble glucans but not by soluble mannans. Three types of experiments demonstrated a role for CR3 in these responses. First, neutrophil ingestion of either yeast or yeast-derived beta-glucan particles was blocked by monoclonal anti-CR3, fluid-phase iC3b, or soluble beta-glucan from barley. Monocyte ingestion of beta-glucan particles was also blocked by anti-CR3, but not by anti CR1 or anti-C3. Second, the neutrophil superoxide burst response to either zymosan or beta-glucan particles was blocked by anti-CR3 or fluid-phase iC3b, and was completely absent with neutrophils from 3 patients with an inherited deficiency of CR3. Third, CR3 was isolated from solubilized neutrophils by affinity chromatography on beta-glucan-Sepharose. PMID- 3040333 TI - Role of complement receptor type three and serum opsonins in the neutrophil response to yeast. AB - Previous studies have suggested that neutrophil complement receptor type three (CR3) has two binding sites: (1) a site for fixed iC3b that does not trigger ingestion or a superoxide (O2-) burst, and (2) a function-triggering site for the beta-glucan component of yeast (Saccharomyces cerevisiae) cell walls. In the present study it was found that yeast (Y) coated with C3b (YC3b) or iC3b (YC3bi), prepared with purified complement in an IgG-free system, were avidly ingested ans stimulated a vigorous O2- burst, whereas sheep erythrocytes (E) bearing C3b or iC3b, were not ingested and did not give an O2- burst. YC3b and YC3bi contained an amount of fixed C3 that was approximately equal to serum-opsonized Y (OY), and produced O2- bursts comparable to OY. Experiments utilizing rabbit F(ab')2 anticomplement receptor type one (anti-CR1) to block fixed C3b binding to CR1, and monoclonal anti-CR3 (MN-41 or OKM1) to block fixed iC3b and Y cell wall binding to CR3, indicated that the O2- burst response to OY was primarily due to fixed iC3b and Y cell wall binding to CR3. Fixed C3b (that represented 33% of the fixed C3 on OY) and IgG anti-Y antibodies that bound to CR1 and Fc receptors, respectively, were found to contribute little to the response. Although YC3b did bind avidly to neutrophil CR1, the results suggested that the O2- burst response to YC3b was triggered after the initial YC3b binding by the secondary attachment of Y cell wall components to CR3. When neutrophils were treated with anti-CR3, 90% of neutrophils bound YC3b (via CR1), but phagocytosis and an O2- burst were completely absent. Similar findings were made with OKM1-treated neutrophils and YC3bi. Responses of OKM1-treated neutrophils were inhibited because only the iC3b binding site of CR3 was ligated by the YC3bi. Thus, fixed C3b or iC3b on Y mediate avid binding of Y to neutrophils via CR1 or the iC3b-binding site of CR3, respectively, but ingestion and an O2- burst response are only triggered when glucans in the Y cell wall secondarily bind to neutrophils via the beta-glucan binding site of CR3. PMID- 3040334 TI - Interception. III: Postcoital luteal contragestion by an antiprogestin (mifepristone, RU 486) in 62 women. AB - The new antiprogestin mifepristone (RU 486) was studied as an emergency postcoital contragestive. An actual pregnancy rate of 1.6% was observed and was related to the actual conception rate. The follow-up rate was 100%. The patterns of onset and duration of the induced menstruation after mifepristone treatment are described. This method provides a good new interceptive technique when the time for use of postcoital steroids or for a postcoital IUD insertion has lapsed. PMID- 3040335 TI - Interception. IV: Failure of mifepristone (RU 486) as a monthly contragestive, "Lunarette". AB - The new antiprogestin mifepristone (RU 486) was studied as a contragestive for continuous fertility control in 24 menstrual cycles. Two pregnancies out of three conceptions continued in spite of antiprogestin treatment. To date, mifepristone at the doses used appears to be inadequate for monthly use. PMID- 3040336 TI - Mechanisms of escape of visna lentiviruses from immunological control. PMID- 3040337 TI - A review of antigenic variation by the equine infectious anemia virus. PMID- 3040338 TI - The influence of cryopreservation on the ultrastructural morphology of human thyroid cells. AB - The purpose of this study was to investigate the effects of the freeze-thaw procedure on the ultrastructural features of human thyroid cells. Four different stages of thyroid cell preparation were compared: (1) fresh surgical tissue, serving as control, (2) cell suspension before freezing, (3) cell suspension after thawing, and (4) monolayer cell culture, obtained from cells after thawing. Electron microscopic examination of cells from each stage showed that the freeze thaw procedure used caused only minor intracellular alterations restricted to mitochondria. Some of these organelles showed low-amplitude swelling or occasionally appeared condensed. These ultrastructural changes were not paralleled by a decrease in the vitality or sensitivity of the cryopreserved cells to stimulating agents. PMID- 3040339 TI - Inflammatory responses to intraocularly injected interleukin 1. AB - Intraocular injection of highly purified human interleukin 1 into the anterior chamber of the rabbit eye resulted in the accumulation of inflammatory cells. Peak cellular infiltration occurred at 4hr and cells were still present at 24h. Examination of ocular blood vessels in IL-1-injected eyes revealed no abnormalities. IL-1 had no effect on the protein content of the aqueous humour confirming that this monokine has no direct action on the blood-aqueous barrier. Comparison of the potency of IL-1 as a chemoattractant with the 5-lipoxygenase product, leukotriene B4 demonstrated that the former agent was more active by several orders of magnitude. PMID- 3040340 TI - Effects of pirazolac on arachidonic acid metabolism in the human synovial system. AB - Six patients suffering from rheumatoid arthritis with massive knee joint effusions were treated with single daily doses of 600 mg pirazolac, a novel non steroidal anti-inflammatory drug, for 3 days. Before the first dose, 3 hours after the second and the third dose, specimens of plasma and synovial fluid were drawn simultaneously. Plasma and synovial fluid concentrations of pirazolac, as determined by HPLC, amounted to 47.9 micrograms/ml and 19.8 micrograms/ml (Day 2) and 55.5 micrograms/ml and 18.7 micrograms/ml (Day 3), respectively. The samples were analyzed for PGE2, LTB4, LTC4 and LTD4 applying various extraction procedures and subsequent radioimmunoassays. PGE2 levels decreased during treatment from 928 pg/ml to 443 pg/ml after the third dose of pirazolac. LTB4 levels were slightly but insignificantly augmented. LTC4 and LTD4 concentrations were below the detection limit prior to and after administration of the drug. PMID- 3040341 TI - Determination of cyclic AMP and cyclic GMP in psoriatic epidermis and dermis. PMID- 3040342 TI - [Clinical analysis of 94 cases of invasive mole]. PMID- 3040343 TI - Longitudinal differentiation of metaphase chromosomes of Indian muntjac as studied by restriction enzyme digestion, in situ hybridization with cloned DNA probes and distamycin A plus DAPI fluorescence staining. AB - The longitudinal differentiation of metaphase chromosomes of the Indian muntjac was studied by digestion with restriction enzymes, in situ hybridization with cloned DNA probes and distamycin A plus DAPI (4'-6-diamidino-2-phenylindole) fluorescence staining. The centromeric regions of chromosomes 3 and 3 + X of a male Indian muntjac cell line were distinct from each other and different from those of other chromosomes. Digestion with a combination of EcoRI and Sau3A revealed a pattern corresponding to that of C-banding. Digestion with AluI, EcoRII or RsaI yielded a band specific to the centromeric region only in chromosomes 3 and 3 + X. Furthermore, HinfI digestion yielded only a band at the centromeric region of chromosome 3, whereas DA-DAPI staining revealed a single band limited to the extreme end of the C-band heterochromatin of the short arm of 3 + X. These results suggest that centromeres of Indian muntjac chromosomes contain at least four different types of repetitive DNA. Such diversity in heterochromatin was also confirmed by in situ hybridization using specific DNA probes isolated and cloned from highly repetitive DNA families. Heterozygosity between chromosome homologs was revealed by restriction enzyme banding. Evidence is presented for the presence of nucleolus organizer regions (NORs) on the long arm of chromosome 1 as well as on the secondary constrictions of 3 and 3 + X. PMID- 3040345 TI - [Technic of converting quartz to cristobalite in dental investment]. PMID- 3040344 TI - Alsactide: ACTH-agonist for use in microdoses in brain-adrenal and other feedsidewards. AB - An increasing number of ACTH-related peptides have been isolated and/or chemically synthesized. In addition to the multiplicity of molecules, there is experimental evidence for multiple target cells and multiple receptors, and hence for different biological activities. The heptadecapeptide analogue alsactide (ACTH 1-17: Synchrodyn) has a C-terminal amide group (butylamide) and the substitution of beta-alanine for serine in position 1 and of lysine for arginine in position 17. These modifications account for enhanced biological activity and uniquely demonstrated chronopharmacological properties. In adult healthy men, the tailoring of dosage and timing of peptide administration was successful in a selective and transient stimulation of glucocorticoid secretion, without a change in the plasma concentrations of aldosterone and testosterone. The dose of 10 micrograms alsactide injected subcutaneously at awakening is proposed for clinical application with the aim of enhancing cortisol secretion in diurnally active subjects. It is noteworthy that injection of even much higher doses for several days at this particular circadian stage did not elicit detectable antibodies to the peptide in more than 200 patients. Restoring or reinforcing the circadian ordering of bioperiodicities correlated with the adrenocortical cycle can thus be achieved as a result of a time-specified therapy with an ACTH-agonist analogue. The results of studies in experimental animals and of preliminary trials in human beings have emphasized the value of alsactide as the first chronizer-peptide in clinical medicine. Its use in the dosage here proposed is expected to be beneficial for preventing deterioration of the circadian system in the elderly, for enhancing psycho-physical performances, and for gaining compliance with regard to chronic disease as well as tolerance to a number of potentially damaging xenobiotics. PMID- 3040346 TI - [Experimental study of the blood supply of the mandibular ramus using radionuclides]. PMID- 3040347 TI - [Experimental study of a model of acute incomplete cerebral ischemic and its characteristics]. PMID- 3040348 TI - [Pathogenesis of schizophrenia: assay of PG and cAMP in the CSF and plasma in 25 cases]. PMID- 3040349 TI - [Experience in the diagnosis and surgical treatment of 110 cases of insulinoma]. PMID- 3040350 TI - [Chemodectoma: report of 5 cases and review of Chinese literature]. PMID- 3040351 TI - [Establishment of hybridoma cell lines secreting monoclonal antibodies against Enterovirus 70]. PMID- 3040352 TI - [Viral etiological study of the 1983 epidemic keratoconjunctivitis in Chengdu]. PMID- 3040353 TI - [Advantage of the measurement of multiple parameters for pleural effusion for the differential diagnosis of tuberculosis and carcinomatous pleurisy]. PMID- 3040354 TI - [Ultrastructure research on lung small cell carcinoma]. PMID- 3040355 TI - Flow cytometric evaluation of anti-herpes drugs. AB - A rapid means of screening drugs for toxicity and anti-herpes simplex virus activity was developed based on the flow cytometric detection of HSV induced changes in cellular DNA content. Subconfluent monolayers of human diploid fibroblasts (HEL 299) were assayed for DNA content with propidium iodide 24 h after infection with HSV-1 (multiplicity of infection 1-10) and treatment with the drug to be tested. Infection was detected by a broadening of the normal diploid and tetraploid peaks and presence of greater than 4-n DNA staining. Inhibition of viral DNA synthesis and maintenance of the normal growth pattern of control cells was indication of antiviral activity. Toxicity of the compound was indicated by the loss of S phase and tetraploid cell populations. Using this assay, we evaluated the activities of one experimental and two established antiviral agents. PMID- 3040356 TI - A small carcinoma of the rectum with a singular histopathologic feature. Report of a case. AB - A case of small carcinoma of the rectum with four different histologic types is presented. The carcinoma was a round node (7 X 7 X 2 mm) surrounded by a flat discolored zone (14 X 14 mm) and had four independently different histologic types: well-differentiated adenocarcinoma, adenocarcinoma with microtubular structure, mucinous carcinoma, and signet-ring cell carcinoma. In part of the tumor the direct transition between well-differentiated adenocarcinoma and signet ring cell carcinoma was seen. The authors believe that the case of rectal carcinoma was a primary form of linitis plastica. PMID- 3040358 TI - [Mechanism of the effect of irradiation on the activity of the Na+,K+-pump in snail neurons]. PMID- 3040359 TI - [Specific cleavage of the Ha-ras oncogene with topoisomerase I]. PMID- 3040360 TI - [Localization of the gene for the Na+,K+-ATPase alpha subunit on chromosomes of the American mink]. PMID- 3040357 TI - Effect of misoprostol on histamine-stimulated acid secretion and cyclic AMP formation in isolated canine parietal cells. AB - Isolated canine parietal cells were used to study the ability of misoprostol to inhibit acid secretion in the presence of a number of acid secretagogues. Misoprostol inhibited histamine-stimulated acid secretion in a dose-dependent and noncompetitive manner. A concentration of 2-3 X 10(-9) M misoprostol inhibited maximal histamine-stimulated acid secretion by one half. Misoprostol had little to no effect on acid secretion stimulated by carbachol and dibutyryl cAMP, had no effect on the acid secretion directly stimulated by pentagastrin, and only modestly inhibited acid secretion stimulated by forskolin. Misoprostol noncompetitively inhibited cAMP formation in response to histamine, with an IC50 value similar to that for the inhibition of histamine-stimulated acid secretion. These results indicate that: (1) misoprostol specifically inhibits histamine stimulated acid secretion in parietal cells, and (2) the antisecretory action of misoprostol is closely related to the reduction of histamine-stimulated cAMP formation with the site of major action most likely in the coupling process between histamine H2 receptor sites and histamine-sensitive adenylate cyclase. PMID- 3040361 TI - [Effect of v-sis-oncogene on transgenic mice]. PMID- 3040362 TI - [Drosophila mobile element jockey is a retroposon and encodes the GAG-specific protein sequence characteristic for retroviruses]. PMID- 3040363 TI - [Interaction of lexitropsin with DNA: negative result of the attempt to detect complex AT/GC-specific binding]. PMID- 3040364 TI - [Characteristics of the protein specifically binding to the major late promoter of adenovirus type 2]. PMID- 3040365 TI - [Antibodies against S-100 protein cause depolarization of the pyramidal neuron membrane and block synaptic transmission in hippocampus slices]. PMID- 3040366 TI - [Specificity of acid DNAses from marine organisms to local conformation of B DNA]. PMID- 3040367 TI - [Restriction endonuclease SsoII: interaction with modified substrates]. PMID- 3040368 TI - Interaction of clavulanic acid, sulbactam and cephamycin antibiotics with beta lactamases. AB - The inhibitory effects of clavulanic acid, sulbactam and cephamycin antibiotics on chromosomally-mediated or plasmid-mediated beta-lactamases were investigated. The inhibition constants were determined by a non-linear regression analysis. Clavulanic acid and sulbactam had high affinities for the purified plasmid mediated beta-lactamases such as SHV-1, TEM-1 and PSE-4, and were potent inhibitors as shown by their low Ki values. Except for Bacteroides beta lactamase, which is sensitive to inhibition by cephamycin antibiotics, clavulanic acid and sulbactam were found not to be as effective against chromosomally mediated beta-lactamases. The cephamycin antibiotics were better inhibitors of chromosomally-mediated beta-lactamases than those that are plasmid mediated. Except for P99 beta-lactamase, against which sulbactam and clavulanic acid were inactive, the cephamycin antibiotics were less effective inhibitors than sulbactam and clavulanic acid. PMID- 3040369 TI - [Therapy of severe hypertension]. PMID- 3040370 TI - [Electron microscopic demonstration of calicivirus-like particles in the feces of diarrheic calves]. PMID- 3040371 TI - [BHV-1 infection of cattle: stabilization by active immunization of seropositive animals with inactivated vaccine--a field trial]. PMID- 3040372 TI - Cyclandelate as a calcium modulating agent in rat cerebral cortex. AB - Cyclandelate is clinically effective in a variety of cerebrovascular indications, but its precise mode of action is unclear. Hence, this study investigated the interaction of cyclandelate, cyclandelate alcohol and cyclandelate acid with the binding sites for radioactively labelled 3H-nitrendipine, a Ca++ entry blocker of the 1,4-dihydropyridine type, on rat cerebral cortex membranes. Cyclandelate showed a dissociation constant (Kd) of 7.1 +/- 1.4 X 10(-5) mol/L (35% inhibition of 3H-nitrendipine binding at 2 X 10(-4) mol/L cyclandelate), cyclandelate alcohol had a Kd value of 1.7 +/- 0.1 X 10(-4) mol/L (maximal 70% inhibition of 3H-nitrendipine binding) whereas cyclandelate acid was inactive. For comparison, nifedipine (Kd of 2.6 +/- 0.3 X 10(-9) mol/L inhibition of 68% of 3H-nitrendipine binding), d-cis diltiazem (Kd of 1.1 +/- 0.1 X 10(-7) mol/L enhancement of 39% of 3H nitrendipine binding) and +/- -verapamil [Kd values of 1.4 +/- 0.4 X 10(-7) mol/L (38% inhibition) and 5.3 +/- 1.7 X 10(-4) mol/L (62% inhibition)] were used. Thus, cyclandelate may exert its clinical activity in cerebral ischaemia or hypoxia at least in part through a calcium modulatory effect. PMID- 3040373 TI - Nitrates. Mode of action at a cellular level. AB - The nitrates used therapeutically in angina pectoris and congestive heart failure are, from a chemical point of view, organic nitroesters. Their principal pharmacological effect is vascular smooth muscle relaxation, leading to vasodilation, which explains their therapeutic effects. Several mechanisms have been proposed for their mode of action at the cellular level, in order to explain vascular smooth muscle relaxation. Today, there is strong evidence that organic nitroesters stimulate the enzyme guanylate cyclase in the smooth muscle cell. This enzyme produces a cyclic nucleotide, cyclic guanosine-3',5'-monophosphate (cGMP), which in turn eventually lowers the free calcium concentration in the cytosol to induce relaxation. The exact mechanism by which the organic nitroesters stimulate guanylate cyclase is still obscure. Preliminary results from our laboratory indicate that there may be more than one mechanism responsible for the activation of the enzyme. Knowledge of the mode of action at the cellular level is probably important in order to understand the mechanism(s) behind the development of tolerance towards the organic nitroesters. PMID- 3040374 TI - Nitrate tolerance from a biochemical point of view. AB - Tolerance to nitrate vasodilators appears to be a general phenomenon that encompasses all known drugs belonging to this group, with the possible exception of molsidomine, for which tolerance has not yet been unambiguously proven. The mechanism behind tolerance development is still obscure, although decreased distribution of drug to the target tissue (i.e. the vascular wall) may be important. In addition, the production of cyclic guanosine-3',5'-monophosphate (cGMP) [the alleged mediator of nitrate-induced vascular smooth muscle relaxation] is reduced in tolerant tissue, while its degradation is increased. These changes could be due to a direct effect on the enzymes involved in the formation and degradation of cGMP in the cell, i.e. guanylate cyclase and phosphodiesterase, respectively. Furthermore, there is some evidence that the degradation of organic nitroesters in the vascular wall is reduced in tolerant tissue. This could result in a reduced production of unstable chemical intermediates (e.g. nitrosothiols), which have been suggested to act as mediators of guanylate cyclase. PMID- 3040375 TI - Development of adenosine 5'-triphosphate sulfurylase and adenosine phosphosulfate kinase in rat cerebrum and liver. AB - The development of the enzymes of sulfate activation, ATP-sulfurylase and APS kinase has been studied in rat cerebrum and, for comparison, in rat liver as well. Cerebrum contains a species of ATP-sulfurylase which can be inhibited by phenylalanine. The activity of this form of ATP-sulfurylase is high in oligodendroglial cells and parallels the rate of myelination. PMID- 3040376 TI - Identification and localization of glucagon-like peptide-1 and its receptor in rat brain. AB - The existence and distribution of glucagon-like peptide-1 (GLP-1) and its receptor in rat brain in relation to that of glucagon were examined. The concentration of GLP-1 immunoreactivity (GLP-1-IR), measured by a specific and sensitive RIA established in this study with anti GLP-1 serum (LMT-01), was found to be highest in the thalamus-hypothalamus, followed by the medulla oblongata. The distribution of glucagon-like immunoreactivity was similar to that of GLP-1 IR. However, appreciable glucagon immunoreactivity was detected only in the thalamus-hypothalamus. Gel filtration analysis showed the presence of GLP-1-IR of various molecular weights in the extract of thalamus-hypothalamus including that eluted at the same position as synthetic GLP-1 (1-37); moreover, HPLC analysis also confirmed the presence of GLP-1-IR, eluted at the exact position as synthetic GLP-1 (1-37). The distribution of receptors for GLP-1 corresponded with that of GLP-1-IR in the rat brain, except in the pituitary gland. The distribution of these receptors was also similar to that of glucagon receptors. The thalamus-hypothalamus, pituitary gland, and medulla oblongata were rich in GLP-1 and glucagon-binding sites. The binding affinities of GLP-1 and glucagon were in the nanomolar range [disocciation constant Kd approximately equal to 4 nM]. The presence of specific, high affinity receptors for GLP-1 was confirmed by demonstrating that GLP-1 stimulated cAMP formation in the thalamus-hypothalamus and the pituitary gland. The concentration of GLP-1 required for half-maximal stimulation of cAMP formation in these regions was about 1 nM. These results suggest that GLP-1 may be synthesized in certain parts of the brain and play a role as a neurosignal transmitter. PMID- 3040377 TI - Plasma adrenocorticotropin is more sensitive than transcortin production or thymus weight to inhibition by corticosterone in rats. AB - After adrenalectomy, ACTH, corticosterone-binding globulin (CBG), and thymus wet weight increase in rats as a consequence of the removal of corticosterone (B) and decrease again in response to replacement with glucocorticoids. We have studied the effect of replacing adrenalectomized male rats with a variety of different concentrations of B. Plasma concentrations of ACTH and CBG and thymus wet weights were related to the measured concentration of free ultrafilterable B in plasma. In other experiments, the clearance of [125I]CBG was determined in adrenalectomized rats with and without B replacement, and the time required for the changes in plasma CBG concentrations after removal and/or replacement of B in adrenalectomized rats was determined. Five to 7 days after adrenalectomy and institution of a relatively constant B replacement signal, plasma CBG concentrations were highly correlated with circulating B concentrations (r2 = 0.745; P less than 0.001). The effect of B was on the CBG production rate, since clearance did not change. Because CBG concentrations decrease as total B concentrations increase, there is an amplification of free B concentrations with increasing total B. The relationships of plasma ACTH and CBG and thymus wet weight to circulating free B levels showed that 50% inhibition of ACTH was achieved at a free B concentration of 0.8 +/- 0.05 nM, whereas 50% inhibition of CBG and thymus wet weight were achieved at free B concentrations of 4.6 +/- 0.9 and 4.4 +/- 0.6 nM, respectively. These inhibition values correlate well with the known Kd values for the high affinity type I B receptor (0.5 nM) and for the lower affinity type II glucocorticoid receptor (2.5-5 nM), respectively, suggesting that ACTH secretion, CBG production, and thymus wet weight are regulated by the association of B with these receptor types. PMID- 3040378 TI - Detailed kinetic analysis of adrenocorticotropin secretion by dispersed rat anterior pituitary cells in a microperifusion system: effects of ovine corticotropin-releasing factor and arginine vasopressin. AB - We have developed a microperifusion system in which we have examined the ACTH secretory responses of acutely dispersed normal rat anterior pituitary cells to ovine CRF (oCRF) and arginine vasopressin (AVP), alone and in combination. The system approached square-wave stimulus hydrodynamics. ACTH secretion was observed within 5 sec of exposure to either secretagogue and reached a maximum within 20 40 sec. ACTH secretion remained constant for as long as oCRF was perifused and then fell gradually toward the basal level. Persistent ACTH release after oCRF perifusion was stopped could not be explained by persistence of oCRF in the perifusion chamber. In contrast to the response to oCRF, ACTH secretion fell progressively toward basal despite continued AVP perifusion. AVP had a synergistic effect with oCRF only if it was perifused simultaneously with oCRF or within 30 sec after oCRF was stopped; it had no synergistic effect if perifused immediately before oCRF. Simultaneous perifusion of oCRF and AVP resulted in an oCRF-like response of greater magnitude, whereas sequential perifusion of oCRF followed by AVP resulted in the usual plateau response to oCRF followed by the initial spike response characteristic of very high concentrations of AVP alone and a subsequent rapid decrease in secretion despite continued perifusion of AVP. The different kinetic response profiles suggest that oCRF and AVP act via different intracellular signal transduction pathways, and the time and sequence dependency of their synergism suggests that the factors that mediate their interactions have different intracellular half-lives. The microperifusion system appears to be uniquely suited to detailed kinetic analysis of anterior pituitary hormone secretion and the intracellular pathways through which secretagogues act and interact. PMID- 3040379 TI - Specific receptor-mediated stimulation of progesterone secretion and cGMP accumulation by rat atrial natriuretic factor in cultured human granulosa-lutein (G-L) cells. AB - The effect of rat atrial natriuretic factor (ANFIV102-126) on the regulation of steroidogenesis in human G-L cells have been studied in vitro. The treatment of cells with ANF (1 X 10(-8)M) resulted in a 3- to 4-fold increase in progesterone secretion compared to the controls. ANF in combination with LH (200 ng/ml) increased progesterone secretion more than six-fold over controls. Concomitantly, ANF stimulated the accumulation of cGMP 30-fold, whereas the level of cAMP was either unchanged or fell slightly (10-15%). Mono[125I]-iodo-ANF bound to G-L cell surface specific receptors with a Kd of 1.8 X 10(-10)M at a density (Bmax) of 160,000-190,000 sites/cell. The binding of [125I]-iodo-ANF was competed by unlabeled ANF in a dose-dependent manner, and hormones unrelated to ANF, such as angiotensins, ACTH or LH, were unable to compete with [125I]-iodo-ANF. The results indicate that ANF can stimulate progesterone secretion and cGMP accumulation in ovarian G-L cells without affecting the level of cAMP and that the stimulatory effects of ANF on G-L cell steroidogenesis may proceed via mechanism(s) involving an intracellular messenger other than cAMP. PMID- 3040380 TI - Further evidence for a central stimulatory action of catecholamines on adrenocorticotropin release in the rat. AB - Catecholamines may stimulate ACTH secretion during stress. To investigate the nature and site of such an action, plasma ACTH was measured in four groups of unanesthetized adult female rats with an indwelling carotid cannula. Sequential 300-microliter blood samples were taken 60 min, 30 min, and immediately before an intracerebroventricular (icv) infusion of 2.5 microliter adrenaline or noradrenaline and 5, 15, 45, 60, and 120 min after the infusion. The four groups were: 1) intact rats; 2) rats infused 7 days after undergoing a discrete bilateral lesion of the ventral noradrenergic ascending bundle caused by 6 hydroxydopamine, which depleted their hypothalamic adrenaline and noradrenaline levels by 90% and 80%, respectively; 3) rats infused 30 min after pretreatment via the icv route with either prazosin or propranolol; and 4) rats infused 16 and 2 h after two successive intracarotid injections of an anti-rCRH-41 serum. In another group, the effects of icv catecholamine administration were compared with those of an intracerebral (ic) microinfusion close to a single paraventricular nucleus (PVN). Finally, in two additional groups blood was sampled at the above mentioned times before and after a 2-min ether inhalation by intact rats or prazosin- and/or propranolol-pretreated rats. In the intact rats (group 1), a stress-like stimulatory dose response was noted after both adrenaline and noradrenaline infusions, with a half-maximal effect at concentrations of about 0.6 nmol and a maximal effect at 2.7 nmol or more. At maximally effective doses, adrenaline was significantly more active than noradrenaline. In the rats with ventral noradrenergic ascending bundle lesions (group 2), 2.7 nM adrenaline or noradrenaline stimulated ACTH release as in the controls without lesions. In group 3, prazosin blocked the ACTH responses to both adrenaline and noradrenaline, whereas propranolol only blocked the response to adrenaline. In group 4, i.e. rats pretreated with an anti-rCRH-41 serum, the amplitude of the ACTH surge after icv adrenaline or noradrenaline infusion was halved. A unilateral ic catecholamine microinfusion next to the PVN (half the icv dose given in group 1) led to a rapid ACTH release that peaked at half the response measured in the icv infused rats. Ether stress-induced ACTH release was decreased by 50-60% after icv pretreatment with 1 or 10 micrograms prazosin, 1 or 6.5 micrograms propranolol, or a combined dose comprising 1 microgram of both. The following conclusions were reached.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3040381 TI - High osmolality: a potent parathyroid hormone secretogogue in dispersed parathyroid cells. AB - We have examined possible mechanisms by which osmolality might modulate PTH secretion in dispersed bovine parathyroid cells. Increasing medium osmolality by adding sodium chloride causes a marked, dose-dependent increase in PTH release. The maximum effect (4-fold increase) is observed when osmolality is around 650 mosM, with half-maximal stimulation at about 400 mosM. When osmolality is increased to a similar extent with either sucrose or sodium chloride, PTH secretion is enhanced to a comparable degree, suggesting that osmolality itself, rather than ionic strength, is responsible for the increase in PTH secretion. Time course experiments show that the increased secretion of PTH with high osmolality occurs very rapidly (in less than 5 min). In contrast to the suppressive effects of high Ca2+ on PTH release, increasing calcium concentration in the incubation media does not inhibit the stimulation of PTH secretion by high osmolality. Moreover, the effects of dopamine (10(-5) M) and high osmolality on PTH release are additive, further suggesting that high osmolality and cAMP modulate PTH release by different mechanisms. In fact, direct measurement of cellular cAMP in cells exposed to high osmolality shows no change relative to control cells incubated with normal osmolality, 127 +/- 20 vs. 146 +/- 21 fmol/10(5) cells, respectively. Cytosolic Ca2+ increases from 257 +/- 29 nM to 703 +/- 50 nM after 200 mM NaCl is added to the incubation medium at low Ca2+ (0.5mM). Prior removal of extracellular calcium abolished this effect. Increasing the osmolality to a similar level by adding sucrose to the medium does not demonstrate any increase in cytosolic calcium. We conclude that high osmolality is a potent secretogogue in dispersed bovine parathyroid cells. Unlike dopamine and isoproterenol, high osmolality does not act through changes in intracellular cAMP. It also prevents the normal inhibitory effect of high Ca2+ on PTH release. Change of cytosolic Ca2+ is variable and suggests that the effect of high osmolality on PTH release cannot be explained by cytosolic Ca2+ alone. Further understanding of the mechanisms by which osmolality affects PTH release, therefore, may provide clues to the unusual inverse relationship between extracellular and cytosolic calcium and PTH release. PMID- 3040382 TI - Characterization of the insulin receptor kinase from human erythrocytes. AB - The insulin receptor from human erythrocytes was studied for receptor-associated tyrosine kinase activity. Receptor phosphorylation was performed by incubation of the receptor with [gamma-32P]ATP and Mn2+ in the presence or absence of insulin, and the phosphorylated receptor was identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis. In Triton X-100-solubilized, and partially purified, receptor preparations, insulin stimulated the phosphorylation of a 95,000 dalton protein in a dose-dependent fashion. Immunoprecipitation with antiinsulin receptor antibodies indicates that this 95,000 dalton protein corresponds to the beta-subunit of the insulin receptor. Phosphoaminoacid analysis revealed that 32P incorporation occurred predominantly on tyrosine residues of the beta-subunit. In addition, the insulin receptor kinase catalyzed the phosphorylation of Histone H2b. Dose response curves for insulin-stimulated phosphorylation of the beta-subunit and Histone H2b were sigmoidal. Both half maximal effects were observed at 3 X 10(-9) M insulin with maximal effects at 10( 6) M. When insulin binding was examined under the same conditions as phosphorylation, the dose-response curves for receptor occupancy and the kinase activation were nearly superimposable, indicating few or no spare receptors for this response to insulin. Insulin-stimulated receptor phosphorylation was diminished by treatment with N-ethylmaleimide, indicating that the receptor possesses sulfhydryl groups, which are important for its enzyme activity. This study demonstrates that insulin receptor kinase from human erythrocytes shares characteristics which are similar to those found in other classical insulin target cells and tissues. PMID- 3040383 TI - Modulation of catechol estrogen synthesis by rat liver microsomes: effects of treatment with growth hormone or testosterone. AB - The ability of GH from various mammalian species, administered to normal mature male rats by constant infusion, to decrease the hepatic 2-hydroxylation of estradiol (E2) to female levels, as measured by the release of 3H2O from [2 3H]E2, was determined. Rat and human GH (hGH) showed the highest activity while ovine GH was inactive. PRL (0.6 IU/h X kg) administered together with hGH (0.02 IU/h X kg) did not antagonize the feminizing action of GH. Infusion of hGH into male rats decreased the affinity of estradiol 2-hydroxylase for its steroid substrate and altered the linear Lineweaver-Burk plot towards a nonlinear hyperbolic plot characteristic of the female. The apparent Michaelis-Menten constant (Km) for the reaction was 1.69 microM for males and 2.75 microM for testosterone-treated ovariectomized females. An equal mixture of liver microsomes from male and female rats gave kinetic values similar to those observed with males alone. Neonatal imprinting with androgen did not alter the magnitude of the response of female rats to treatment with testosterone and/or GH at maturity and the androgen effect could only be shown in ovariectomized animals. The results with rats of different endocrine status were corroborated by the kinetic data and by the pattern of metabolites obtained with [4-14C]E2 when examined by TLC and autoradiography. The hormonal control of estradiol 2-hydroxylase, the key enzyme in catechol estrogen formation, and the contribution of sex-specific multiple forms of the enzyme to this reaction are discussed. PMID- 3040384 TI - Phosphoinositides turnover and insulin secretion in pancreatic islets. AB - The relationship between glucose-induced insulin secretion and metabolism of inositol phospholipid was investigated by means of an islet perifusion method and direct measuring of inositol phosphates after sonicating the islets. The results showed that the time course of inositol phospholipid breakdown is coincident with the first phase of glucose-induced insulin secretion. Analysis of the effluent perifusate as well as the water soluble inositol-containing substance after sonication of stimulated islets revealed that most of the metabolite of inositol phospholipid is inositol-triphosphate, the hydrolysis product of phosphatidylinositol-4,5-bisphosphate. On the other hand, perifusion of islets with exogenous inositol-triphosphate showed a monophasic and dose-dependent response of insulin secretion. Thus, the initial process of glucose stimulation is accompanied with the formation of inositol-triphosphate, which is a possible candidate for the triggering of first phase insulin secretion. PMID- 3040385 TI - A case of hyponatremia in panhypopituitarism caused by the primary empty sella syndrome. AB - A 64-year-old woman was admitted for evaluation of hyponatremia. She was maintained on hypertonic saline administration. Without this therapy, the serum Na concentration decreased progressively to 127 mEq/L and the plasma osmolality to 254 mOsm/Kg H2O, on Day 3. At that time, the concentration of antidiuretic hormone (ADH) was as high as 3.5 pg/ml. A skull radiogram revealed an enlarged sella turcica. Computed tomography (CT) revealed a low density in the sella, and magnetic resonance imaging revealed equal intensity of the sella turcica and the cerebrospinal fluid. A diagnosis of empty sella syndrome was made by metrizamide cisternography in conjunction with CT scanning. A diagnosis of panhypopituitarism was made by endocrine function tests. 123I-thyroidal uptake was 6% when her serum TSH was 10.9 microU/ml, suggesting that she might also have primary hypothyroidism. When this patient was given glucocorticoid before levothyroxine replacement, her serum Na concentration rose up to about 140 mEq/L and a normal relationship between her plasma ADH level (2.4 pg/ml) and plasma osmolality (281 mOsm/kg H2O) was restored. Therefore, it was suggested that ADH hypersecretion induced by the glucocorticoid deficiency might in part contribute to the development of hyponatremia. This is the case of primary empty syndrome associated with panhypopituitarism, in whom initial symptom was caused by hyponatremia. PMID- 3040386 TI - Corticotropin and angiotensin induce dephosphorylation of a Mr-20,000 protein in bovine adrenal cells. AB - ACTH (1-24) and Angiotensin II, both able to activate steroidogenesis in bovine fasciculata-reticularis cells, each reduced the [32P] incorporation in a cytosolic Mr-20,000 pI 6.8 protein in this cell. Cells preincubated with Sar1 Angiotensin prevented the effect of Angiotensin. Angiotensin 10(-8)M and ACTH 10( 10)M led to an almost complete disappearance of the corresponding radioactive spot on the autoradiograph. The effect was observed as soon as 2 minutes after addition of hormones to the cells. Other activators of steroidogenesis such as 8 bromocyclicAMP (8-BrcAMP), 4 beta-Phorbol-12 beta-Myristate-13 alpha-acetate (PMA) and [9-tryptophan (o-nitrophenylsulfenyl)] substituted ACTH (NPS-ACTH), also reduced the labeling of the Mr-20,000 polypeptide. On the other hand, this effect was not reproduced by insulin or human growth hormone (hGH). On 2-D gels from control, the coincidence of this polypeptide with phosphorylated myosin light chain was not observed. We suggest that the apparent dephosphorylation of this polypeptide may represent a common effect of all steroidogenic agents regardless of their seemingly distinct early actions. PMID- 3040387 TI - Neural plasticity vs. disease. Focus on the NMDA receptor in epilepsy and mental disorders. Beverly Hills, California, U.S.A., March 30-31, 1987. Abstracts. PMID- 3040388 TI - Equine parvovirus--a cause for concern? PMID- 3040389 TI - Total parenteral nutritional therapy of a foal with diarrhoea from which parvovirus-like particles were identified. PMID- 3040390 TI - Activation of sodium transport in human erythrocytes by beta-adrenoceptor stimulation in vivo. AB - Beta-adrenoceptor stimulation in vivo shifts potassium into the cells. To examine whether human erythrocytes participate in this process, we measured, along with serum or plasma potassium, the concentrations of potassium and sodium in erythrocytes. Beta-adrenoceptor stimulation was obtained by infusion of either fenoterol or hexoprenaline into 6 volunteers at rest or by endogenous amines provoked in 14 volunteers during ergometric exercise. Metabolic effects were followed at rest on serum insulin, C-peptide, and growth hormone levels, and during exercise on pH on lactate concentration in blood. The potassium concentration (mean +/- S.E.M.) dropped (p less than 0.01) in serum from 4.64 +/- 0.37 to 3.19 +/- 0.43 mmol x l-1 in the first hour at rest and in plasma from 5.70 +/- 0.93 to 4.63 +/- 0.45 in 90 sec directly after exercise. The concentration of erythrocyte sodium dropped (p less than 0.001) from 9.68 +/- 0.73 to 8.81 +/- 0.62 mmol x l-1 in cells and from 9.62 +/- 1.16 to 8.55 +/- 1.24 during exercise for 90 s, respectively. Changes in the concentration ratio of cellular sodium to potassium confirmed this sodium shift. An increased sodium transport in erythrocytes due to beta-adrenoceptor stimulation in vivo appears to complement a shift of serum potassium into the cells and may be mediated by the membrane-bound sodium, potassium ATPase. PMID- 3040391 TI - The effect of exposure to high and low frequency hand-arm vibration on finger systolic pressure. AB - Twenty-three patients with hand-arm vibration exposure and diagnosed vibration syndrome were given a thorough clinical and neurophysiological examination, together with finger strain gauge plethysmography. Eleven of the patients were forest workers regularly using chain saws (low frequency vibration exposure), and twelve were metal grinders (higher frequency vibration exposure). Both groups had significantly lower finger blood pressures than healthy controls, and comparisons between the groups indicated that the mean values tended to be lower in the grinders. The findings suggest that hand-arm vibration exposure is associated with obstructive changes in the distal arteries of the fingers, and that vibration frequency is one of the factors determining the severity of the changes and the time of onset of the symptoms. PMID- 3040393 TI - Prevalence of antibodies to herpes simplex virus type 1 in different population groups in Italy. PMID- 3040394 TI - Purealin, a novel activator of skeletal muscle actomyosin ATPase and myosin EDTA ATPase that enhanced the superprecipitation of actomyosin. AB - 1. Purealin, a novel bioactive principle of a sea sponge Psammaplysilla purea, activated the superprecipitation of myosin B (natural actomyosin) from rabbit skeletal muscle. The maximum change in the turbidity increased with increasing purealin concentrations and was three times the control value in the presence of 50 microM purealin. 2. The ATPase activity of myosin B was also elevated to 160% of the control value by 10 microM purealin. On the other hand, purealin inhibited the myosin ATPase in the presence of 10 mM CaCl2 and 0.5 M KCl (Ca2+-ATPase), and the concentration for the half inhibition was 4 microM. 3. On the other hand, purealin activated the myosin ATPase in the presence of 5 mM EDTA and 0.5 M KCl (EDTA-ATPase). The maximum activation by 10 microM purealin was 160% of the control value. 4. Furthermore, similar results concerning the modification of ATPase activities by purealin were obtained in myosin subfragment-1 instead of myosin. 5. These results suggest that purealin activates the superprecipitation of myosin B by affecting the myosin heads directly. It is also an interesting observation that there is a correlation between the activities of the myosin EDTA ATPase and actomyosin ATPase of myosin B. PMID- 3040395 TI - Reactions of nucleic acid bases with alpha-acetylenic esters. Chemical modification of poly(A) and poly(C). AB - The reaction of chlorotetrolic (4-chloro-2-butynoic) esters with adenine and cytosine derivatives, in which a new heterocycle bearing an alkylating chloromethyl side chain is fused to the purine or pyrimidine ring, was extended to poly(A) and poly(C) used as models of nucleic acids. The derivatization proceeds under mild conditions and its extent can be controlled by the reaction time. The additional rings can exist in two isomeric forms and the nature of the isomer formed depends on steric factors in the vicinity of the reacting base. The reaction with chlorotetrolic esters discriminates between the single-stranded (reactive) and double-stranded (unreactive) forms, between the exposed an hidden adenine and cytosine bases and even between the exposed and sterically hindered fragments of the base moiety and thus allows structural investigations of these nucleic acids. The chloromethyl group of the derivatized nucleobases can be used to bind the modified polymers to other molecules. PMID- 3040396 TI - EcoR124 and EcoR124/3: the first members of a new family of type I restriction and modification systems. AB - We have purified the EcoR124 and EcoR124/3 restriction enzymes and shown that they are type I enzymes by several criteria: subunit composition, DNA and S adenosylmethionine-dependent ATPase activity, and site-specific DNA methylase activity. By immunochemical criteria these enzymes are related to each other but are unrelated to the two previously investigated families of type I restriction enzymes. They form therefore a new family which we call type IC. The arrangement of the structural genes coding for these enzymes and their transcriptional organisation have been determined. These are different from the common arrangement found for the other two families of type I enzymes. PMID- 3040392 TI - Perinatal viral infections. AB - In comparison to older children and adults, neonates are immunologically incompetent. They are susceptible to infections caused by a variety of microorganisms, including bacteria, fungi and viruses. These infectious agents may be acquired by neonates either prenatally, during the intrapartum period or postnatally. The purpose of this review is to emphasize the potential impact of viral infections contracted by neonates at the time of delivery or within the neonatal period. The viruses reviewed include the herpes group of viruses (cytomegalovirus, herpes simplex viruses and varicella-zoster virus), type B hepatitis virus, human immunodeficiency virus, respiratory viruses, enteroviruses, rotavirus and human papilloma virus. For each virus the potential sources and incidence of the infection, the common manifestations of the illness, and possible means of prevention and therapy are discussed. Although infections caused by bacteria tend to be more clinically dramatic and more immediately life threatening, it is emphasized that infections caused by viruses are common and associated with substantial long-term morbidity. Perinatal viral infections need to be recognized as early in life as possible so that their natural history can be more completely defined and any possible intervention made. PMID- 3040397 TI - The gamma-aminobutyric-acid/benzodiazepine-receptor protein from rat brain. Large scale purification and preparation of antibodies. AB - The gamma-aminobutyric-acid-receptor protein complex from rat brain was solubilized in high yield, purified in milligram amounts by benzodiazepine affinity chromatography and used to generate a high-titer rabbit antiserum. High concentrations of Triton X-100 detergent plus KCl solubilized about 90% of the membrane-bound gamma-aminobutyric acid receptor (assayed by [3H]muscimol binding) and benzodiazepine receptor (assayed by [3H]flunitrazepam binding) activities. Both activities were retained on an affinity column using an immobilized benzodiazepine ligand, and most of the column-absorbed receptor could be eluted by a solution of free benzodiazepine plus 4 M urea. The purified protein bound [3H]muscimol and [3H]flunitrazepam with receptor-like pharmacological specificity and specific activities of about 1700 pmol and 700 pmol bound/mg protein, respectively, for the two ligands. This corresponds to a purification of over 600 fold and a near theoretical purity, with a yield of milligram quantities from 100 g brain. Four peptide bands were observed on gel electrophoresis in sodium dodecyl sulfate, with molecular mass values of 31, 47, 52 and 57 kDa. The latter two were most significantly stained, and identified as receptor subunits by photolabeling with [3H]flunitrazepam (52 kDa) and [3H]muscimol (57 kDa), and by reaction on Western blots with monoclonal antibodies to this protein produced by Schoch et al. [(1985) Nature (Lond.) 314, 168-171]. Rabbit antiserum was raised to the purified protein and could, at high dilutions, both coprecipitate soluble gamma-aminobutyric-acid/benzodiazepine-receptor-binding activities and stain the receptor subunits (principally 52-kDa band) on Western blots. PMID- 3040398 TI - Identification of protein phosphatases-1 and 2A and inhibitor-2 in oocytes of the starfish Asterias rubens and Marthasterias glacialis. AB - Protein phosphatases present in the particulate and soluble fractions of oocytes of the starfish Asterias rubens and Marthasterias glacialis have been classified according to the criteria used for these enzymes from mammalian cells. The major protein phosphatase activity in the particulate fraction had very similar properties to protein phosphatase-1 from mammalian tissues, including preferential dephosphorylation of the beta subunit of phosphorylase kinase, sensitivity to inhibitor-1 and inhibitor-2, inhibition of phosphorylase phosphatase activity by protamine and heparin, and retention by heparin Sepharose. The major protein phosphatase in the soluble fraction had very similar properties to mammalian protein phosphatase-2A, including preferential dephosphorylation of the alpha subunit of phosphorylase kinase, insensitivity to inhibitors-1 and 2, activation by protamine and heparin, and exclusion from heparin-Sepharose. An acid-stable and heat-stable protein was detected in the soluble fraction of starfish oocytes, whose properties were indistinguishable from those of inhibitor-2 from mammalian tissues. It inhibited protein phosphatase-1 specifically, and its apparent molecular mass on SDS polyacrylamide gels was 31 kDa. Furthermore, an inactive hybrid formed between the starfish oocyte inhibitor and the catalytic subunit of mammalian protein phosphatase-1 could be reactivated by preincubation with MgATP and mammalian glycogen synthase kinase-3. The remarkable similarities between starfish oocyte protein phosphatases and their mammalian counterparts are indicative of strict phylogenetic conservation of these enzymes. The results will facilitate further analysis of the role of protein phosphorylation in the control of starfish oocyte maturation by the hormone 1-methyladenine. PMID- 3040399 TI - Hormonal regulation of carbamoyl-phosphate synthetase I synthesis in primary cultured hepatocytes and Reuber hepatoma H-35. Defective regulation in hepatoma cells. AB - Regulation of carbamoyl-phosphate synthetase I (CPS) synthesis by various hormones was compared in primary cultured hepatocytes from adult rat and in Reuber hepatoma H-35 by pulse labeling of the cells with [35S]methionine. CPS synthesis in hepatocytes was stimulated 8-fold and 5-fold by dexamethasone and glucagon respectively. CPS synthesis in hepatocytes was synergically (about 50 fold) stimulated by a combination of dexamethasone and glucagon. Less synergic stimulation was observed by combining dexamethasone with N6, O2' dibutyryladenosine 3',5'-monophosphate (dibutyryl-cAMP) or with isoproterenol. The basal level of CPS synthesis in hepatoma cells was higher than that in hepatocytes. CPS synthesis in hepatoma cells was stimulated by dexamethasone and dibutyryl-cAMP but the extent was only 3-fold and 1.8-fold respectively. The synergic effect of combination of dexamethasone and dibutyryl-cAMP was not observed in hepatoma cells. Neither glucagon nor isoproterenol exhibited an appreciable effect on CPS synthesis in hepatoma cells. Insulin and epinephrine suppressed CPS synthesis both in hepatocytes and hepatoma cells. The effect of epinephrine was indicated to be through alpha-adrenergic receptors. The effects of insulin and epinephrine were additive on CPS synthesis both in hepatocytes and hepatoma cells. PMID- 3040400 TI - Isolation of a cDNA and characterization of the 5' flanking region of the gene encoding the type I regulatory subunit of the cAMP-dependent protein kinase. AB - A bovine cDNA probe for the type I regulatory subunit of the cAMP-dependent protein kinase [Lee et al. (1983) Proc. Natl Acad. Sci. USA 80, 3608-3612] was used to screen two lambda gt11 libraries constructed from poly(A)-rich RNA from the porcine kidney cell line, LLC-PK1. A series of overlapping clones were isolated and characterized. The largest clone, lambda RI15, of 1426 bp was found to code for the entire RI protein but was apparently missing the 3' end of the mRNA. The porcine cDNA codes for a protein of 389 amino acids that shows 99% homology to bovine RI and hybridizes to two major mRNA transcripts of approximately 2.0 kb and 4.5 kb from LLC-PK1 cells. The porcine cDNA for RI was used to screen a genomic library of LLC-PK1 DNA constructed in the EMBL-3 vector and several clones were isolated and characterized. By using a probe from the 5' end of the RI cDNA we isolated the 5' end of the gene and 700 bp of the promoter region of the gene were sequenced. The promoter region lacks a characteristic TATA box but contains two inverted CAAT boxes and is rich in G + C residues. Several sequence motifs were identified in the 5' promoter region which could be responsible for the regulation of synthesis of this gene. Multiple transcription initiation sites were identified by S1 nuclease mapping. PMID- 3040401 TI - The elucidation of the microheterogeneity of highly purified p-hydroxybenzoate hydroxylase from Pseudomonas fluorescens by various biochemical techniques. AB - Highly purified p-hydroxybenzoate hydroxylase from Pseudomonas fluorescens can be separated into at least five fractions by anion-exchange chromatography. All fractions exhibit the same specific activity and the enzyme exists mainly in the dimeric form in solution. Sodium dodecyl sulfate/polyacrylamide gel electrophoresis of a mixture of the different fractions reveals two apparent forms of enzyme molecules, while isoelectric focusing experiments, on the other hand, reveal six apparently different forms of enzyme molecules. It is shown that the different forms of enzyme molecules are due to the (partial) oxidation of Cys 116 in the sequence of the enzyme. This interpretation of the data is supported by kinetic measurements of the formation of hybrid dimeric molecules monitored by fast protein liquid chromatography, using purified enzyme containing Cys-116 either in the native and or the fully oxidized (sulfonic acid) state. By chemical modification studies using maleimide derivatives, 5,5'-dithiobis(2-nitrobenzoate) and H2O2, it is shown that sulfenic, sulfinic and sulfonic acid derivatives of Cys-116 are products of oxidation. The results are briefly discussed with respect to the possibility that this isolation artifact might also be partially responsible for the appearance of multiple forms of enzyme molecules in other biochemical preparations. PMID- 3040402 TI - Nickel-[iron-sulfur]-selenium-containing hydrogenases from Desulfovibrio baculatus (DSM 1743). Redox centers and catalytic properties. AB - The hydrogenase from Desulfovibrio baculatus (DSM 1743) was purified from each of three different fractions: soluble periplasmic (wash), soluble cytoplasmic (cell disruption) and membrane-bound (detergent solubilization). Plasma-emission metal analysis detected in all three fractions the presence of iron plus nickel and selenium in equimolecular amounts. These hydrogenases were shown to be composed of two non-identical subunits and were distinct with respect to their spectroscopic properties. The EPR spectra of the native (as isolated) enzymes showed very weak isotropic signals centered around g approximately 2.0 when observed at low temperature (below 20 K). The periplasmic and membrane-bound enzymes also presented additional EPR signals, observable up to 77 K, with g greater than 2.0 and assigned to nickel(III). The periplasmic hydrogenase exhibited EPR features at 2.20, 2.06 and 2.0. The signals observed in the membrane-bound preparations could be decomposed into two sets with g at 2.34, 2.16 and approximately 2.0 (component I) and at 2.33, 2.24, and approximately 2.0 (component II). In the reduced state, after exposure to an H2 atmosphere, all the hydrogenase fractions gave identical EPR spectra. EPR studies, performed at different temperatures and microwave powers, and in samples partially and fully reduced (under hydrogen or dithionite), allowed the identification of two different iron-sulfur centers: center I (2.03, 1.89 and 1.86) detectable below 10 K, and center II (2.06, 1.95 and 1.88) which was easily saturated at low temperatures. Additional EPR signals due to transient nickel species were detected with g greater than 2.0, and a rhombic EPR signal at 77 K developed at g 2.20, 2.16 and 2.0. This EPR signal is reminiscent of the Ni-signal C (g at 2.19, 2.14 and 2.02) observed in intermediate redox states of the well characterized Desulfovibrio gigas hydrogenase (Teixeira et al. (1985) J. Biol. Chem. 260, 8942]. During the course of a redox titration at pH 7.6 using H2 gas as reductant, this signal attained a maximal intensity around -320 mV. Low temperature studies of samples at redox states where this rhombic signal develops (10 K or lower) revealed the presence of a fast-relaxing complex EPR signal with g at 2.25, 2.22, 2.15, 2.12, 2.10 and broad components at higher field. The soluble hydrogenase fractions did not show a time-dependent activation but the membrane-bound form required such a step in order to express full activity.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3040403 TI - Nucleotide sequence and characterization of the yeast PSS gene encoding phosphatidylserine synthase. AB - 1. A yeast chromosomal DNA which contains the structural gene for phosphatidylserine synthase (PSS) was isolated by genetic complementation from a wild-type yeast genomic library. The PSS gene was subcloned into a 1.1-kb fragment of the yeast DNA on the YEp13 vector. 2. The PSS gene on the multicopy plasmid caused the fourfold over-production of the enzyme and fully restored the phosphatidylserine content of the transformant. The phospholipid composition of the transformant was similar to that of the wild type. 3. Sequence analysis showed that this DNA fragment contains an open reading frame capable of encoding 276 amino acid residues with a calculated relative molecular mass of 30,804. Northern blot analysis of poly(A)-rich RNA of the wild-type yeast indicated that this DNA segment is transcribed into a single mRNA species. 4. The DNA sequence contained two putative transcriptional initiation signals, each followed by the ATG initiator codon. Deletion experiments indicated that the 5'-proximal ATG codon is essential for the synthesis of the functional phosphatidylserine synthase. PMID- 3040404 TI - The Noc region of Ti plasmid C58 codes for arginase and ornithine cyclodeaminase. AB - Plant tumors induced by Agrobacterium tumefaciens synthesize a group of substances (opines) which can serve as sole source of carbon and nitrogen for the bacteria. We investigate Ti-plasmid-coded genes and enzymes involved in catabolism of the opine N2-(1,3-dicarboxypropyl)-L-arginine (nopaline) with a novel approach: expression and mapping of protein-coding regions in Escherichia coli minicells, followed by identification of enzyme functions in the heterologous E. coli background. The results show that a specific part of the nopaline catabolism (Noc) region of Ti plasmid C58 is packed with closely spaced protein-coding regions which can be expressed into polypeptides of distinct sizes in E. coli. We identify and map three enzyme activities: nopaline oxidase, arginase and ornithine cyclodeaminase, an unusual protein converting ornithine directly into proline. Nopaline oxidase requires two different Noc-gene-encoded proteins for function and the latter two enzymes are new discoveries in the Noc region. These three enzyme activities together constitute a catabolic pathway leading from nopaline through arginine and ornithine to proline. PMID- 3040405 TI - Comparison of binding sites in DNA for berenil, netropsin and distamycin. A footprinting study. AB - Techniques of DNase I and micrococcal nuclease footprinting have been used to compare the binding sites for berenil, netropsin and distamycin on two different DNA fragments. Each ligand binds to the A + T-rich zones which contain clusters of at least four A.T base pairs. Neither guanosine nor cytidine nucleotides appear to be allowed within the A + T-rich runs which constitute the preferred binding sites, although they are sometimes protected from DNase I cleavage in neighbouring regions. Berenil and netropsin share with distamycin the property of causing enhanced rates of cleavage at certain sequences flanking their binding sites. There are significant differences in the concentrations of each ligand required to produce defined patterns of protection, seemingly dependent upon the nature (and possibly the gross base composition) of the piece of DNA being used in the experiment. PMID- 3040406 TI - Construction and stability of a sixfold repeated artificial gene. AB - The concatenation of an artificial gene consisting of 69 base pairs is described. A simple and versatile procedure was employed which is applicable to any DNA segment bordered by different restriction sites. If applied in the 'continuous duplication mode', it does not necessitate the isolation of any DNA fragment. The addition of adaptor oligonucleotides to one terminus of the segment is required to create complementary ends. The ligation of single-stranded oligonucleotides to single-stranded DNA termini was analyzed and optimized. A simple procedure is described for the separation of excess adaptor molecules. The in vivo stability of constructs containing up to six directly repeated genes was investigated. Up to four copies of the gene appeared stable during transformation of strain HB101. With six copies exact excision of ony copy was occasionally observed after transformations of rec+, recA, and recBC host cells. During long-term growth the hexameric construct appeared stable in all three strains. PMID- 3040407 TI - Protein phosphatases of the guinea-pig parotid gland. AB - The nature of protein phosphatases of the guinea-pig parotid gland was investigated. The protein phosphatases were characterized by (a) the use of five different 32P-labelled substrate proteins (phosphorylase a, histone H2B, casein, and the alpha and beta subunits of phosphorylase kinase), (b) their behaviour during ion-exchange chromatography, (c) their relative molecular mass distribution during gel filtration, (d) their sensitivity towards inhibition by inhibitor 2, (e) their ability to be stimulated by protamine and (f) by their behaviour during freezing and thawing in the presence of 2-mercaptoethanol. The following results were obtained. 1. The 'cytosol' (100,000 X g supernatant) contains protein phosphatases of the types 1, 2A and 2B. 2. On the basis of inhibition with inhibitor 2 (1.2 micrograms/ml) the 'cytosolic' phosphorylase phosphatase activity consists to about 40% of protein phosphatase 1 and to about 60% of protein phosphatase 2A. 3. In the cytosol about 80-90% of the protein phosphatases 1 and 2A exist in an inactive state. 4. A 5-10-fold activation can be achieved by ethanol precipitation, which results in the generation of a mixture of forms of low apparent molecular mass of about 30 kDa. 5. Microsome associated phosphorylase phosphatase activities can be extracted in a highly active state by detergent (1% Triton X-100) or by 0.8 M NaCl. 6. Activity measurements in the presence of inhibitor 2 (1.2 micrograms/ml) indicate that the microsomal activities consist to about 75% of protein phosphatase 1 and to about 25% of protein phosphatase 2A. Activities corresponding to protein phosphatases 2B and 2C could not be detected. 7. The 'microsomal' protein phosphatase activities exhibit lower apparent molecular masses (70 kDa and 30 kDa) than the 'cytosolic' protein phosphatases (about 260 kDa). 8. After ethanol treatment of the microsomal protein phosphatases only activities with apparent molecular masses of about 30 kDa can be detected. These share several similarities with the ethanol-treated cytosolic protein phosphatases. 9. Both cytosolic and microsomal protein phosphatases display activity towards histone H2B and casein. PMID- 3040408 TI - Are calcium-dependent protein kinases involved in the regulation of glycolytic/gluconeogenetic enzymes? Studies with Ca2+/calmodulin-dependent protein kinase and protein kinase C. AB - Changes in glycolytic flux have been observed in liver under conditions where effects of cAMP seem unlikely. We have, therefore, studied the phosphorylation of four enzymes involved in the regulation of glycolysis and gluconeogenesis (6 phosphofructo-1-kinase from rat liver and rabbit muscle; pyruvate kinase, 6 phosphofructo-2-kinase and fructose-1,6-bisphosphatase from rat liver) by defined concentrations of two cAMP-independent protein kinases: Ca2+/calmodulin-dependent protein kinase and Ca2+/phospholipid-dependent protein kinase (protein kinase C). The results were compared with those obtained with the catalytic subunit of cAMP dependent protein kinase. The following results were obtained. 1. Ca2+/calmodulin dependent protein kinase phosphorylates 6-phosphofructo-1-kinase and L-type pyruvate kinase at a slightly lower rate as compared to cAMP-dependent protein kinase. 2. 6-Phosphofructo-1-kinase is phosphorylated by the two kinases at a single identical position. There is no additive phosphorylation. The final stoichiometry is 2 mol phosphate/mol tetramer. The same holds for L-type pyruvate kinase except that the stoichiometry with either kinase or both kinases together is 4 mol phosphate/mol tetramer. 3. Rabbit muscle 6-phosphofructo-1-kinase is phosphorylated by cAMP-dependent protein kinase but not by Ca2+/calmodulin dependent protein kinase. 4. Fructose-1,6-bisphosphatase from rat but not from rabbit liver is phosphorylated at the same position but at a markedly lower rate by Ca2+/calmodulin-dependent protein kinase when compared to the phosphorylation by cAMP-dependent protein kinase. 5. 6-Phosphofructo-2-kinase is phosphorylated by Ca2+/calmodulin-dependent protein kinase only at a negligible rate. 6. Protein kinase C does not seem to be involved in the regulation of the enzymes examined: only 6-phosphofructo-2-kinase became phosphorylated to a significant degree. In contrast to the phosphorylation by cAMP-dependent protein kinase, this phosphorylation is not associated with a change of enzyme activity. This agrees with our observation that the sites of phosphorylation by the two kinases are different. The results indicate that Ca2+/calmodulin-dependent protein kinase but not protein kinase C could be involved in the regulation of hepatic glycolytic flux under conditions where changes in the activity of cAMP-dependent protein kinase seem unlikely. PMID- 3040409 TI - Cycloheximide induces accumulation of vasoactive intestinal peptide (VIP) binding sites at the cell surface of a human colonic adenocarcinoma cell line (HT29-D4). Evidence for the presence of an intracellular pool of VIP receptors. AB - Incubation of monolayers of HT29-D4 cells (a clone of the human colonic adenocarcinoma cell line HT29) in the presence of 17.5 microM cycloheximide resulted in an increase in the number of vasoactive intestinal peptide (VIP) binding sites at the cell surface without any change in the affinity of receptor for its ligand. The increase in 125I-VIP-binding capacity was dose-dependent between 0.35 microM and 17.5 microM cycloheximide and was correlated with the inhibition of protein biosynthesis. At higher concentrations of drug (17.5-100 microM) a plateau corresponding to a twofold increase in VIP-binding capacity was reached independently of the extent of protein synthesis inhibition. We found that VIP receptors of HT29-D4 cells with such an enhanced binding capacity behaved like those of control cells with respect to receptor internalization and recycling (i.e. the cycle of occupied receptors was insensitive to cycloheximide). After inactivation of 90% of cell-surface VIP receptors by alpha chymotrypsin, we observed a biphasic kinetic of reappearance of VIP-binding sites. 40% of VIP-binding sites reappeared very quickly (less than 5 min) and 100% within 17 h. The fast recovery of VIP receptors was probably due to the deployment of new binding sites from an intracellular pool. The rate and extent of recovery of these receptors were similar in control cells and in cycloheximide treated cells. However, the slow recovery was inhibited in cycloheximide-treated cells probably because a pool of immature receptors was depleted by the drug before the alpha-chymotrypsin treatment. Our data are consistent with the existence of two different intracellular pathways of occupied and unoccupied VIP receptors. PMID- 3040410 TI - On the effects of weak bases and monensin on sorting and processing of lysosomal enzymes in human cells. AB - The weak bases chloroquine, primaquine, NH4Cl and the ionophore monensin exert similar but not identical effects on sorting, transport and processing of cathepsin D in several human cell lines (fibroblasts, HepG2 cells, U937, monocytes). The drugs inhibit the segregation of newly synthesized cathepsin D from the secretory route. The kinetics of transport of nonsegregated cathepsin D precursor along the secretory route is retarded resulting in a delayed hypersecretion. Higher concentrations of the drugs can arrest the intracellular transport completely. The extent of inhibition of segregation varies among the different human cell types tested. Thus, in fibroblasts the secretion can be stimulated to exceed 80%, while in U937 cells the secretion cannot be enhanced above 50% although both cell types have the same basal rate of secretion (approximately 10% of the synthesized cathepsin D). We suggest that pH independent sorting mechanisms contribute to the targeting of cathepsin D in U937 cells. Processing of the cathepsin D remaining in cells is characteristically changed depending on the drug. The proteolytic processing is strongly inhibited by chloroquine and is rather insensitive to monensin. Unlike the other drugs, monensin blocks the formation of complex oligosaccharides in cathepsin D and allows for extensive secretion solely of molecules that are sensitive to endo H. PMID- 3040411 TI - Differential response of stress fibers and myofibrils to gelsolin. AB - The actin-severing activity of human platelet gelsolin was analyzed on embryonic skeletal and cardiac myofibrils, and on stress fibers in non-muscle cells. These subcellular structures, although in all three cell types composed of contractile proteins arranged in sarcomeric units, were found to respond differently to gelsolin. The myofibrils in permeabilized myotubes or cardiac cells, as well as in living, microinjected muscle cells proved resistant to a wide concentration range of gelsolin. The same was found for the "mini-sarcomeres" which are seen in developing muscle cells. In contrast, stress fibers in microinjected fibroblasts or epithelial cells, as well as in permeabilized cells, were broken down rapidly by the platelet gelsolin. We conclude from these results that the mini-sarcomeres in embryonic myotubes and cardiac myocytes are not identical with stress fibers. PMID- 3040412 TI - Probing of the structural stability of vimentin and desmin-type intermediate filaments with Ca2+-activated proteinase, thrombin and lysine-specific endoproteinase Lys-C. AB - Intermediate filaments (IFs) reconstituted from purified, delipidated vimentin and desmin as well as respective protofilaments were subjected to degradation by Ca2+-activated neutral thiol proteinase, thrombin and lysine-specific endoproteinase Lys-C, respectively. The breakdown products were analyzed by SDS polyacrylamide gel electrophoresis and negative stain electron microscopy. While Ca2+-activated proteinase and thrombin caused rapid and complete degradation of IFs with kinetics not significantly different from those of the degradation of protofilaments, lysine-specific endoproteinase did not exert any electron microscopically detectable effect on filament structure. Although both types of subunit proteins were truncated at their non-alpha-helical, C-terminal polypeptides by this proteinase, they were still able to assemble into 10 nm filaments. Closer electron microscopic inspection of IFs treated with Ca2+ activated proteinase revealed numerous ruptures along the filaments already at very early stages of digestion. SDS-polyacrylamide gel electrophoresis of the processed filaments in conjunction with previous biochemical characterizations of the breakdown of protofilaments by Ca2+-activated proteinase showed that these inhomogeneities primarily arose from degradation of the arginine-rich, non-alpha helical N-termini of the filament proteins. These findings demonstrate that, although the N-terminus of vimentin and desmin is essential for filament stability, it is still highly susceptible to proteolytic attack in particular and very likely to posttranslational modification in general. Such structural modifications of the N-termini of IF proteins might exert great influences on the intracellular distribution and molecular organization of IFs in various physiological and pathological conditions. PMID- 3040413 TI - Inhibition of fibroblast chemotaxis by superoxide dismutase. AB - Superoxide radicals are known to be important mediators in chronic inflammatory and fibrotic processes, in which accumulation of fibroblasts is thought to play a major role in the pathogenetic events. The enzyme superoxide dismutase removes these radicals by a catalytic reaction. Chemotactic response of human fibroblasts and fibrosarcoma-derived cells (HT-1080) to fibroblast conditioned medium, fibronectin and platelet-derived growth factor was inhibited in a dose-dependent manner in the presence of superoxide dismutase, while random migration, cell proliferation, cell viability and synthesis of collagen and non-collagenous proteins was not altered. In contrast, phorbol myristate acetate, an inducer of superoxide generation, stimulated the chemotactic movement of fibroblasts to the attractants. Evidence for the formation of superoxide is provided by the reduction of tetrazolium salt by activated fibroblasts which could be inhibited by superoxide dismutase. Thus, it is concluded that superoxide in small amounts is involved in the mechanism of fibroblast chemotaxis. Superoxide dismutase may, therefore, reduce fibroblast migration into sites of injury or inflammation. PMID- 3040414 TI - Quantification and localization of galactose-specific binding sites in rat liver during postnatal development. AB - We investigated the number and distribution of galactose-specific binding sites in developing livers from suckling rats of various ages using Lac-BSA-Au5 (lactosylated bovine serum albumin adsorbed onto colloidal gold particles 5 nm in diameter) as electron-dense ligand, and performing transmission electron microscopy of the specimen. It has been reported that the number of galactose specific binding sites increases rapidly during organ development post partum (p.p.) and this was ascribed to hepatocyte receptor increase only. We now have investigated in in situ and in vitro experiments whether the binding sites of identical sugar specificity but located on sinusoidal cells show the same increase in expression or are independently regulated. We therefore quantified the number of particles bound by isolated hepatocytes and liver macrophages and found a gradual increase of both binding activities with age, the binding levels of adult liver cells being reached at day 15 p.p. This was confirmed with experiments using in situ prefixed organs thus proving the validity of this finding also for the intact organ. In both sets of experiments--in vitro as well as in vivo--ligand was found binding statistically distributed as single particles on hepatocytes of all ages, whereas on liver macrophages the binding pattern changed during development. On liver macrophages from rats 15 days of age ligand binding occurs in the preclustered pattern described for macrophages from adult rat livers whereas liver macrophages of newborn rats express a different binding pattern: they bind the ligands mostly as single particles with only few and small microaggregates.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3040415 TI - Relationship between tumour size and uptake of radiolabelled anti-CEA in a colon tumour xenograft. AB - The relationship between tumour size and the uptake of three radiolabelled anti CEA localising antibodies (A5B7, 1H12 and PK2G) into a human colon tumour xenograft (MaWi) has been examined. For tumour weights greater than 100 mg (109 873 mg) there was a strong positive correlation between absolute uptake and tumour weight with mean uptakes per gram of 9.8 (r = 0.92), 5.0 (r = 0.93) and 5.3 (r = 0.94) for A5B7, 1H12 and PK2G respectively. For tumour weights below 100 mg (17-99 mg) the percentage uptake per gram (specific uptake) increased markedly reaching 80% of the injected dose for A5B7. The above phenomena could be modelled by representing uptake by the surface area of a sphere and tumour weight by its volume. Transformation of this model produced a linear relationship suitable for regression analysis of the experimental data. The slopes of the regression lines for the three antibodies were very close to that predicted by the model suggesting that their uptake into MaWi xenografts is proportional to surface area. The main discrepancy of the actual data was shown by the intercepts which relate to the variation in uptake between different antibodies. This model provides a possible means of correcting for the effect of tumour size when investigating the uptake of antibodies into xenografts. PMID- 3040416 TI - Cyclic nucleotides in cerebrospinal fluid of drug-free Parkinson patients. AB - Concentrations of cyclic nucleotides--adenosine-3',5'-monophosphate (c-AMP) and guanosine-3',5'-monophosphate (c-GMP)--were measured in cerebrospinal fluid (CSF) of 17 drug-free Parkinson patients and 12 controls. No significant difference between the cyclic nucleotide contents (p greater than 0.05) in CSF of patients and controls was detected, nor was there a correlation between the content and the degree of neurological disability. Besides, no changes in the cyclic nucleotide contents were detected in the subgroups of patients according to the prominence of tremor or rigidity/akinesia as the main symptoms of the disease. PMID- 3040417 TI - Nonbacterial thrombotic endocarditis and nondisseminated malignancy associated with osteopetrosis. AB - Nonbacterial thrombotic endocarditis (NBTE) is a presentation of an occult malignancy. The clinical, laboratory and postmortem findings of a 44-year-old man with osteopetrosis and NBTE are reported. A small apical bronchogenic carcinoma was found on postmortem examination. A thorough search for malignancy should be made in patients with unexplained cerebral emboli, especially if hematologic abnormalities are present. There is presently no evidence that anticoagulation or other treatment is beneficial. PMID- 3040418 TI - Uncompacted lamellae in three patients with POEMS syndrome. AB - Three patients with POEMS syndrome were studied: two of them had myeloma, whereas the third had M protein without myeloma. Peripheral nerve biopsies showed no deposits of anti-Ig sera at direct immunopathological examination. Ultrastructural study revealed myelino-axonal degeneration in all three cases. This was associated in the first case with a dramatic loss of myelinated fibers and there were lesions consistent with acute degeneration in the second case. In addition, all three had some fibers showing uncompacted myelin lamellae. This peculiar modification has previously been reported only in cases of dysglobulinemia, lymphoma and inflammatory demyelinating polyneuropathy, but its mode of formation remains unclear. PMID- 3040419 TI - Non-steroidal anti-inflammatory drugs: how do they work? AB - Current dogma holds that non-steroidal anti-inflammatory drugs (NSAIDs) act by inhibition of the synthesis and release of prostaglandins. However, NSAIDs also inhibit the activation of neutrophils, which provoke inflammation by releasing products other than prostaglandins. We now report that NSAIDs (for example, indomethacin, piroxicam) inhibit activation of neutrophils by inflammatory stimuli such as C5-derived peptides and leukotriene B4 even when cyclooxygenase products generated in suspensions of stimulated neutrophils (prostaglandin E and thromboxanes) are present. Sodium salicylate (3mM) greatly inhibited aggregation of neutrophils but had no effect on aggregation of platelets or production of thromboxane induced by arachidonate. Sodium salicylate and other NSAIDs also inhibit calcium movements (45Ca uptake, changes in fluorescence of chlortetracycline and Quin-2). Aspirin, sodium salicylate, indomethacin, and piroxicam also enhanced the post-stimulation rise in intracellular cyclic AMP. NSAIDs therefore inhibit early steps in neutrophil enhance intracellular levels of cyclic AMP. PMID- 3040421 TI - Priming of neutrophils for enhanced oxidative burst by sputum from cystic fibrosis patients with Pseudomonas aeruginosa infection. AB - Neutrophils accumulate in the lungs of cystic fibrosis (CF) patients and inflict tissue damage by release of oxygen radicals and proteases. Here we report on the ability of sputum to prime neutrophils for enhanced release of oxygen radicals. Sol phase was prepared by ultracentrifugation of sputum obtained from CF patients attending the CF Clinic, Rigshospitalet, Copenhagen. The oxidative burst response of neutrophils from healthy individuals was measured by oxygen consumption, superoxide production and chemiluminescence. Neutrophils were preincubated with sputum or buffer and then stimulated with f-Met-Leu-Phe or zymosan. Appropriate controls were included in the experiments. It was shown that neutrophils preincubated with CF sol phase and stimulated with f-Met-Leu-Phe generated a three- to five-fold higher chemiluminescence response than those preincubated with buffer. There was no enhancement of the response when zymosan was used for stimulation of the cells. Neutrophils incubated with sol phase alone exhibited no response. Attempts were made to identify and partially characterize the priming factor(s). It was found that the sputum samples contained bacterial endotoxins. The priming activity was resistant to heating at 100 degrees C for 15 min, and was present only in fractions with molecules larger than 100 KD. It is suggested that the priming factor(s) consist of bacterial endotoxins and/or immune complexes. Activation and enhanced release of oxygen radicals from neutrophils may play an important role in the host defence as well as pathogenesis of tissue damage in the lungs of these patients. PMID- 3040420 TI - Complete iodide trapping defect in two cases with congenital hypothyroidism: adaptation of thyroid to huge iodide supplementation. AB - Two cases of congenital defect in iodide trapping mechanism are related. The absence of thyroid and gastric concentration of 99mTcO4 led to the diagnosis. The study of saliva and gastric:serum concentration ratios confirmed the complete defect. The kinetics of radioiodine studied by external detection showed an early simultaneous decay in the thyroid, the stomach and the left ventricle. Thyroid accumulation of 131I, demonstrated by camera imaging, was estimated to be 0.1% at 48 h. It probably originated from simple diffusion. Iodide supplementation was progressively increased to 4.5 g and 10 g day-1 respectively. It resulted in a normalization of all parameters. Huge doses of iodide did not result in any evidence of hyperthyroidism as TSH rose normally after TRH. Intermittent iodide supplementation in one case could not maintain euthyroidism longer than a few weeks. Daily treatment, therefore, seems necessary. PMID- 3040422 TI - ACTH-(1-24) restores blood pressure in acute hypovolaemia and haemorrhagic shock in humans. PMID- 3040423 TI - Selective up-regulation by interferon-gamma of surface molecules of the Ly-6 complex in resting T cells: the Ly-6A/E and TAP antigens are preferentially enhanced. AB - Surface molecules encoded by the murine Ly-6 locus can transduce triggering signals in T cells and thus may play important roles in T cell function. Previously, we found that Ly-6 molecules are up-regulated by interferon (IFN) alpha/beta in resting T cells. Here, we examined the possible influence of IFN gamma on these molecules. Purified T cells from C57BL/6 (Ly-6.2) and BALB/c (Ly 6.1) mice were incubated in vitro with recombinant murine IFN-gamma and the expression of Ly-6 antigens was measured by flow cytofluorometry. It was found that both Ly-6A/E and T cell-activating protein (TAP) molecules are markedly enhanced while Ly-6C is less affected. Under the same conditions, other T cell surface molecules showed no or marginal changes. The effect of IFN-gamma on Ly 6A/E and TAP expression reached a maximum with as little as 10 U/ml and required only 18-24 h of incubation. Moreover, the enhancement of Ly-6A expression induced by IFN-gamma was stable for at least 5 days. Analysis of T cell subsets further revealed that IFN-gamma-induced augmentation of Ly-6A (C57BL/6 mice) involves both Lyt-2+ and L3T4+ cells while the increase of Ly-6E (BALB/c mice) is more pronounced in Lyt-2+ cells. The functional consequence of these phenotypic alterations was evaluated by studying the mitogenic responses of T cells to antibody-mediated Ly-6 cross-linking in the presence of phorbol myristate acetate. Pretreatment of resting T cells with IFN-gamma dramatically increased the responses to anti-Ly-6A and anti-Ly-6E monoclonal antibodies. IFN-gamma treatment also boosted the stimulation induced by anti-TAP monoclonal antibody when this stimulation was performed under suboptimal conditions. Therefore, IFN gamma selectively up-regulates the Ly-6A/E and TAP activation pathways in resting T cells. We speculate that this effect may contribute to the immunoregulatory activities of IFN-gamma. PMID- 3040424 TI - Epstein-Barr virus-transformed human precursor B cell lines: altered growth phenotype of lines with germ-line or rearranged but nonexpressed heavy chain genes. AB - A series of lymphoblastoid cell lines (LCLs) have been established by in vitro infection of fetal bone marrow and fetal liver cells with Epstein-Barr virus (EBV). While most lines showed the usual mature B cell phenotype, a small proportion were cytoplasmic and surface immunoglobulin (Ig) heavy and light chain negative. Analysis of gene rearrangements indicated that the Ig- lines were either germ-line or nonproductively rearranged when probed for JH and were in germ-line configuration for C chi; no mu or chi mRNA could be detected in such cells. Precursor B cell lines were indistinguishable from their normal Ig+ counterparts in their expression of a wide variety of cell surface markers including "activation" antigens usually associated with the lymphoblastoid state; even the single LCL showing germ-line heavy and light chain genes expressed B lineage-specific cell surface antigens. However, the Ig- lines were distinct from their Ig+ counterparts in three important respects: (a) they grew much more slowly and achieved lower saturation densities, (b) they showed unusually high proportions (8-16%) of cells in EBV-productive cycle, and (c) they contained unusually high proportions (up to 40%) of cells expressing free joining (J) chain. These results suggest that precursor B cells differ in their response to the growth-transforming effects of EBV such that the virus-cell interaction in precursor B cell lines is inherently less stable than in conventional LCL. In particular there may be a greater movement of cells out of cycle and along the B cell maturation pathway. It is possible that such movement leads in individual cells either to virus replication or to a "sterile" plasmacytoid differentiation with J chain expression in the absence of Ig synthesis. PMID- 3040425 TI - Receptors for substance P and neurokinins. Correlation between binding and biological activities. AB - The biological activities of neurokinin-related peptides were compared with their binding affinities measured in various laboratories. A positive significant correlation was demonstrated between the relaxation of the dog carotid artery and the binding of Bolton-Hunter [125I]substance P to rat brain synaptosomes, the contractions of the rat duodenum and the rabbit pulmonary artery and the binding of Bolton-Hunter [125I]neurokinin A to duodenum smooth muscle plasma membranes, and the contraction of the rat portal vein and the binding of [125I]Bolton-Hunter NH-senktide to rat cerebral cortex membranes. PMID- 3040426 TI - The specific opioid kappa-agonist U-50,488H inhibits low Km GTPase. AB - DAGO, a specific opioid mu-agonist stimulated the low Km GTPase activity of membranes of the guinea-pig striatum at 1-1000 nM and this effect was antagonized by naloxone 10-100 nM. On the other hand, U-50,488H, a specific opioid kappa agonist inhibited the low Km GTPase activity of membranes of the guinea-pig cerebellum at 1-100 nM. As this effect was antagonized by MR2266 but not by naloxone, the findings suggest that opioid mu- and kappa-receptor stimulation is probably linked to the stimulation and inhibition of low Km GTPase, respectively. PMID- 3040427 TI - High concentrations of naloxone attenuate N-methyl-D-aspartate receptor-mediated neurotoxicity. AB - (-)-Naloxone, 1 mM, partially reduced neuronal loss induced by exposure of murine cortical cell cultures to N-methyl-D-aspartate (NMDA) or quinolinate, but produced little or no attenuation of kainate or quisqualate neurotoxicity. Antagonism of NMDA neurotoxicity was (-)-naloxone concentration-dependent between 100 microM and 3 mM. (+)-Naloxone produced a slightly greater reduction of NMDA neurotoxicity, arguing against mediation by opioid receptors. Although this novel neuron-protective action of (-)-naloxone was weak, it may contribute to reported beneficial effects in CNS ischemia. PMID- 3040428 TI - N-ethylmaleimide blocks the modulatory effects of divalent cations and guanine nucleotides on the brain substance P receptor. AB - The binding of [3H]physalaemin ([3H]PHY) to rat brain substance P receptors is modulated by cations and guanine nucleotides. [3H]PHY binding in the presence of either monovalent or divalent cations (125 mM Na2SO4 or 2.5 mM MnCl2) shows a KD of 5.9 and 5.5 nM and a Bmax of 44.4 and 63.9 fmol/mg protein respectively. In the presence of both, there is a 2-fold increase in the affinity (KD 2.8 nM) and a 25-80% increase in the Bmax (81.6 fmol/mg protein). Addition of 100 microM GTP or Gpp(NH)p in either 125 mM Na2SO4 or 2.5 mM MnCl2 or both decreases the Bmax by 25-55%. However, the receptor affinity for [3H]PHY is not significantly altered by guanine nucleotides. N-Ethylmaleimide (NEM) irreversibly inhibits the receptor binding with an IC50 of 1.0 mM, demonstrating that SH groups play a critical role in the interaction of the ligand with the receptor. If the SP receptors are protected with 1 microM PHY, NEM irreversibly inhibits the effect of divalent cations and guanine nucleotides. Analysis of [3H]PHY binding in 125 mM Na2SO4, 2.5 mM MnCl2 on membranes that were protected with 1 microM PHY and then preincubated with NEM demonstrates a variable decline in receptor number and a 2 fold decrease in the affinity (KD, from 2.8 to 6.9 nM). These observations indicate the existence of a second class of SH groups that are essential for the interaction of divalent cations and guanine nucleotides with the receptor. The blockade of the modulatory effects of divalent cations and guanine nucleotides by NEM treatment further suggests that brain SP receptors are coupled to a guanine nucleotide binding regulatory protein. PMID- 3040429 TI - Chronic clonidine treatment produces desensitisation of post- but not presynaptic alpha 2-adrenoceptors. AB - Rats were treated chronically with clonidine by osmotic minipump (500 micrograms/kg daily for 14 days). One day after the end of the infusion period, the animals were pithed, and alpha 2-presynaptic and -postsynaptic adrenoceptor responses evaluated using inhibition of cardioaccelerator response to sympathetic stimulation by guanabenz, and pressor response to guanabenz respectively. Chronic clonidine treatment had no effect on the presynaptic effect of guanabenz, but the postsynaptic response to this agonist was decreased. In a separate group of pithed rats treated chronically with clonidine, there was no alteration in the increment in plasma noradrenaline concentration produced by sympathetic stimulation, and no alteration in the yohimbine-induced enhancement of plasma noradrenaline response to sympathetic stimulation. These data argue against desensitisation of peripheral presynaptic alpha 2-adrenoceptors by chronic administration of alpha 2-adrenoceptor agonist. Sympathetic hyperactivity following withdrawal of chronic clonidine treatment may be mediated by down regulation of postsynaptic alpha 2-adrenoceptors in the CNS. PMID- 3040430 TI - Regulation of high- and low-affinity [3H]imipramine recognition sites in rat brain by chronic treatment with antidepressants. AB - Specific binding of [3H]imipramine to its recognition sites in frontal cortex and levels of serotonin (5-HT), 5-hydroxyindoleacetic acid (5-HIAA) and norepinephrine (NE) as well as uptake of serotonin by crude synaptosomal (P2) fraction were determined in a group of rats chronically (for 21 days) treated with different types of antidepressant drugs: nortriptyline, fluoxetine, iprindole, phenelzine (10 mg/kg per day), maprotiline (20 mg/kg per day) and vehicle only (controls). Quantitative analysis of imipramine competition curves confirmed the existence of two classes of [3H]imipramine sites: high-affinity with IC50 and 11.2 nM and low-affinity with IC50 of 630 nM for the competing ligand. The proportion of high- and low-affinity sites was 73 +/- 4 and 26 +/- 4%, respectively. Chronic treatment with all antidepressant drugs except iprindole significantly decreased the affinity but not the proportion of high affinity sites for imipramine. IC50 of imipramine at low-affinity sites was even more markedly increased at low-affinity sites by all treatments except for phenelzine. Fluoxetine was by far the most effective in altering the affinity of both high- and low-affinity [3H]imipramine recognition sites. Both NE and 5-HT levels were significantly enhanced only by phenelzine treatment, whereas 5-HT and 5-HIAA levels were found to be lower after fluoxetine. Kinetics of 5-HT uptake were altered significantly only in rats treated with fluoxetine: rate of 5-HT uptake (Vmax) was decreased by 43% and Km value increased from 104 to 184 nM. Changes in affinity of imipramine for its binding sites were not found to be associated with the effect of tested drugs on 5-HT levels or uptake. They may be due to adaptive alterations in physico-chemical properties of binding proteins although the presence of residual drug interfering with the binding assay cannot be excluded. The observed changes are likely to represent the condition during chronic administration of these drugs in clinical therapy of depression. PMID- 3040431 TI - Alpha 2-adrenoceptor modulation of nociception in rat spinal cord: location, effects and interactions with morphine. AB - The effects of intrathecal clonidine alone and prior to intrathecal morphine were studied on electrically evoked A beta and C fibre activity in the dorsal horn of the halothane-anaesthetised rat. Clonidine reduced C fibre-evoked activity in a dose-dependent manner, to a maximum 52% inhibition which was reversed by rauwolscine and yohimbine but not naloxone. High doses of clonidine also produced small inhibitions of A fibre-evoked activity. Clonidine potentiated the inhibitory action of intrathecal morphine on electrically evoked C fibre activity but not A fibre activity. In addition, the location of alpha 2-adrenoceptor and opiate binding sites in consecutive sections of rat lumbar cord was investigated using in vitro autoradiography with selective ligands, and it was demonstrated that both opiate and alpha 2-receptor types are present within the same superficial layers of the dorsal horn of the same animal. The results indicate that alpha 2-adrenoceptors and opiate receptors can interact in the modulation of nociceptive transmission in rat spinal cord. PMID- 3040432 TI - Adrenomedullary and beta-adrenergic participation in enhanced sympathetic pressor responses of spontaneously hypertensive rats. AB - The beta-adrenergic and adrenomedullary components of pressor responses to sympathetic nerve stimulation were studied in spontaneously hypertensive rats (SHR) and normotensive Wistar Kyoto rats (WKY). The effects of electrical stimulation of the entire spinal cord of pithed rats pretreated with tubocurarine and atropine were studied on systolic blood pressure, heart rate and plasma cyclic AMP levels. The heart rate increase upon low frequency stimulation (1 Hz) and the blood pressure elevation upon stimulation at higher frequencies (3 and 5 Hz) were higher in SHR than in WKY whereas the increase in circulating cyclic AMP level was not different in the two strains. Pretreatment with propranolol (2.5 mg X kg-1) further enhanced the pressor responses in SHR but not in WKY, although it inhibited the heart rate acceleration and decreased the circulating level of cyclic AMP similarly in the two strains. After acute adrenalectomy, the elevations of blood pressure and circulating cyclic AMP levels were reduced to an identical level in SHR and WKY. These results show that the marked enhancement of the pressor response observed in SHR upon stimulation of the entire sympathetic outflow is mostly of adrenomedullary origin and includes a hypotensive component due to beta-adrenoceptor stimulation which is not present in WKY. PMID- 3040433 TI - Benzodiazepine receptors studied in living primates by positron emission tomography: inverse agonist interactions. AB - The convulsant actions of methyl 6,7-dimethoxy-4-ethyl-beta-carboline-3 carboxylate (DMCM) and of methyl beta-carboline-3-carboxylate (beta-CCM) were evaluated in the baboon (Papio papio). DMCM, 0.6-4 mg/kg, induced epileptic seizures with short latency. DMCM convulsive seizures could be blocked by i.v. administration of the benzodiazepine agonist diazepam (10 mg). Similarly, beta CCM, 0.3-3 mg/kg i.v., provoked generalized seizures in the baboons. These seizures were also reversed by the administration of propyl beta-carboline-3 carboxylate (3 mg/kg) or of diazepam (5 mg/kg). Combining the results from Positron Emission Tomography and the EEG assessments, benzodiazepine receptor occupancy by beta-CCM and DMCM was directly correlated with their convulsant actions in the living baboon. beta-CCM exerted its convulsant action in the living baboon at 76 and 74% benzodiazepine receptor occupancy in, respectively, occipital and temporal cortices whereas DMCM displayed a similar convulsive activity when only 58 and 65% of these receptors in the above regions were occupied. PMID- 3040434 TI - Non-stereospecific excitatory actions of morphine may be due to GABA-A receptor blockade. AB - An explosive motor behavior (EMB) similar to that seen following morphine injection into the rat periaqueductal gray (PAG) was observed following an injection of GABA-A receptor antagonists into the rat PAG. In general, the potencies of certain opiates and known GABA-A antagonists in producing EMB following their injections into the PAG paralleled their potencies as GABA antagonists in a radioreceptor assay. We suggest that one of the dual actions of morphine in the CNS may be GABA blockade. PMID- 3040435 TI - Peripheral benzodiazepine binding sites on platelet membranes are increased during diazepam treatment of anxious patients. AB - Reduced (24%) [3H]PK 11195 binding capacity to platelet membranes was observed in anxious patients in comparison to age- and sex-matched normal controls. Four weeks of diazepam treatment induced elevation (69%) in the maximal number of binding sites, and 1 week of drug withdrawal resulted in a slight decrease (16%) in 'peripheral' benzodiazepine binding sites as compared to their level during treatment. PMID- 3040436 TI - Functional evaluation of glutamate receptor subtypes in cultured cerebellar neurones and astrocytes. AB - Micromolar concentrations of kainic acid and quisqualic acid released [3H]D aspartate preaccumulated by cerebellar granule cells in culture. The effect of kainate was selectively antagonized by kynurenic acid and, less effectively, by PDA. Kainate and quisqualate also increased [3H]GABA release from a subpopulation of cultured cerebellar astrocytes. Kynurenic acid selectively blocked the effect of kainic acid. NMDA and no D-aspartate or GABA releasing effect. These results suggest the existence of two different excitatory amino acid sites active on neurotransmitter amino acid release in both cerebellar granule cells and astrocytes. PMID- 3040437 TI - Norepinephrine modulates seizures induced by quinolinic acid in rats: selective and distinct roles of alpha-adrenoceptor subtypes. AB - We investigated in rats whether alterations in noradrenergic function caused by 6 hydroxydopamine or alpha- and beta-adrenoceptor agonists and antagonists would modify the susceptibility of the brain to electroencephalographic seizures induced by intrahippocampal infusion of quinolinic acid. 6-Hydroxydopamine depletion of norepinephrine facilitated the expression of seizures while alpha adrenoceptor stimulation by clonidine had either proconvulsant (0.1 mg/kg) or anticonvulsant (from 0.5 to 2 mg/kg) effects. Clonidine's anticonvulsant activity (0.5 mg/kg) was mimicked by methoxamine given intrahippocampally (10 micrograms), and antagonized by prazosin (1 mg/kg), whereas both yohimbine (5 and 10 mg/kg) and piperoxane (5 mg/kg) had no significant effect. Seizure facilitation induced by clonidine (0.1 mg/kg) was blocked by yohimbine (10 mg/kg). Systemic (0.25 and 0.5 mg/kg) or intrahippocampal (10 and 20 micrograms) isoproterenol and propranolol (10 mg/kg) had no effect. Spiking activity and neurotoxicity induced by quinolinic acid were unaltered by treatments which protected against convulsions. Modulation of quinolinic acid-convulsive activity by alpha adrenoceptor subtypes appears to be selective and complex, since alpha 1-type activation reduces seizures while alpha 2-type stimulation has proconvulsant effects. PMID- 3040438 TI - Behavioural evidence for a selective GABA-A antagonistic activity of SR 95103 and SR 42641 after intrastriatal injection in mice. AB - Two pyridazinyl GABA derivatives, SR 95103 and SR 42641 have recently been described as selective GABAA receptor antagonists. We have now investigated the behavioural effects of SR 95103 and SR 42641 after intrastriatal injection in mice. When injected into the right striatum, SR 95103 (0.01-0.5 microgram), SR 42641 (0.0001-0.01 microgram) and bicuculline methiodide (0.005-0.05 microgram) induced contralateral rotations which were antagonized by intraperitoneal injection of muscimol. In contrast, the intrastriatal injection of the GABAA receptor agonist muscimol induced ipsilateral rotations. Muscimol-induced turning was antagonized by SR 95103 (10-30 mg/kg), SR 42641 (1-10 mg/kg) and (+) bicuculline (0.125-0.5 mg/kg) injected intraperitoneally, but not by strychnine. Intrastriatal glycine also induced ipsilateral rotations which were antagonized by strychnine (0.01-0.3 mg/kg i.p.) but not by (+)-bicuculline, SR 95103 or SR 42641. These results suggest that SR 95103 and SR 42641 induce turning through a selective blockade of GABAA receptors within the striatum. PMID- 3040439 TI - Pharmacological properties of a proenkephalin A-derived opioid peptide: BAM 18. AB - BAM 18 is a derivative of the opioid precursor proenkephalin A. Although it exists in rat and guinea-pig brain in relatively high concentrations, its physiological function is presently unknown. In the present study we have determined the opioid receptor selectivity of this peptide using radioligand binding and peripheral tissue bioassay. When selective binding conditions were used, BAM 18 bound to the mu opioid receptor with an affinity three times that of the kappa opioid receptor and over 10 times that of the delta opioid receptors (Ki = 0.29, 0.75, and 3.2 nM respectively). BAM 18 also displayed mixed receptor selectivity in in vitro bioassay. Ke values for naloxone antagonism of BAM 18 agonist activity in the electrically stimulated guinea-pig ileum and the mouse vas deferens were 4.3 and 9.9 nM, respectively. These data indicate that BAM 18 binds to all three opioid receptor subtypes with a selectivity profile of mu greater than kappa greater than delta. PMID- 3040441 TI - Pharmacologic profile of chemically stable analogs of peptide leukotrienes. AB - Peptide leukotrienes are potent constrictors of airway smooth muscle but lack chemical stability. Replacement of the natural triene backbone of leukotrienes with 9-(x-heptylphenyl)-7-nonenoic acid (x = 2, 3, or 4) renders these analogs chemically stable and pharmacologically active. The para, meta and ortho substitutions of the heptyl (C7H15) moiety were combined with the different leukotriene peptide substitutions of glutathionyl, C-peptide of leukotriene C4, cysteinylglycinyl, D-peptide of leukotriene D4, and cysteinyl, E-peptide of leukotriene E4 rendering nine active analogs. The pharmacology of these analogs was evaluated in isolated guinea-pig tracheal strips. The para-substituted C peptide analog was the most potent; however, it was 100-fold less potent than leukotriene C4. The contractile activities of the para- and meta-substituted C peptide analogs were enhanced by L-serine borate (45 mM), indicating they were substrates for gamma-glutamyl transpeptidase. FPL55712 (10 microM) failed to antagonize the substituted C-peptide analogs independent of the presence of L serine borate. The contractile activities of the three substituted D-peptide analogs were enhanced by L-cysteine, indicating they are all substrates for aminopeptidase. The para-substituted D-peptide analog was the most potent D peptide analog, but it was 10- and 1,000-fold less active than the para substituted C-peptide analog and leukotriene D4, respectively. The para- and meta substituted E-peptide analogs were approximately 1,000-fold less potent than leukotriene E4, and like the substituted D-peptide analogs, they were antagonized by FPL55712. In contrast, the ortho-substituted E-peptide analog was devoid of intrinsic activity but antagonized leukotriene E4-induced contractions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3040442 TI - The effect of two novel dipeptide antagonists of excitatory amino acid neurotransmission on the high pressure neurological syndrome in the rat. AB - Intracerebroventricular injection in the rat of beta-D-aspartyl aminomethylphosphonate (Asp-Amp) 1 mumol, or Y-D-glutamylaminomethylsulphonate (GAMS) 1 mumol, increases the onset pressure for the initial tremor phase of the high pressure neurological syndrome (HPNS) by 50%. Asp-Amp also significantly increases the onset pressures for myoclonus and for tonic-clonic seizures. GAMS did not significantly change the onset pressures for myoclonus or tonic clonic seizures, but it caused the appearance of brief clonic seizures prior to the onset of the HPNS. PMID- 3040440 TI - Microvascular leakage to platelet activating factor in guinea-pig trachea and bronchi. AB - Conscious guinea-pigs received platelet activating factor (PAF, 0.03-0.25 microgram/kg, i.v.) and colloidal carbon (C, tracer for microvascular leakage). Fifteen min later the animal was killed and C-labelled microvessels (leakage) were detected in the mucosal/submucosal region of tracheal and bronchial sections. PAF was more potent than LTD4 or histamine. The numbers of leaky vessel were dose-related, arterioles and arteries were not affected, leaky vessels were seen from proximal trachea to intrapulmonary bronchi and carbon was not apparent in the alveolar wall. The effect was quick in onset, of short duration, could be repeated, was not produced by lyso-PAF and was unaffected by thrombocytopaenia (produced by an antiserum). Thrombocytopaenia itself did not cause leaky vessels in the airways nor affect histamine responses. Thus, PAF causes an increase in microvascular leakage in the conducting airways, which is not dependent on platelets. The affected vessels are probably postcapillary bronchial venules and PAF may act via an endothelial cell receptor in these microvessels. This leakage effect of PAF in the airways could contribute to features of bronchial asthma. PMID- 3040443 TI - Neuropeptide Y modulates adrenergic neurotransmission by an endothelium dependent mechanism. PMID- 3040444 TI - Both the sigma receptor-specific ligand (+)3-PPP and the PCP receptor-specific ligand TCP act in the mouse vas deferens via augmentation of electrically evoked norepinephrine release. PMID- 3040445 TI - Increase in IP3 precedes alpha-adrenoceptor-induced increase in force of contraction in cardiac muscle. AB - In electrically driven left auricles isolated from rat hearts the alpha 1 adrenoceptor agonist phenylephrine increased inositol trisphosphate (IP3) and force of contraction. The increase in IP3 preceded the increase in force of contraction, indicating that the alpha 1-adrenoceptor-mediated increase in IP3 and force of contraction may have been causally related. PMID- 3040446 TI - Chronic haloperidol treatment up-regulates rat brain PCP receptors. PMID- 3040447 TI - Protein disulfide crosslinking stabilizes a polyoma large T antigen-host protein complex on the nuclear matrix. AB - We have investigated the effects of intermolecular disulfide crosslinking and temperature-dependent insolubilization of nuclear proteins in vitro on the association of the polyoma large T antigen with the nuclear matrix in polyomavirus-infected mouse 3T6 cells. Nuclear matrices, prepared from polyomavirus-infected 3T6 cells by sequential extraction of isolated nuclei with 1% Triton X-100 (Triton wash), DNase I, and 2 M NaCl (high salt extract) at 4 degrees C, represented 18% of total nuclear protein. Incubation of nuclei with 1 mM sodium tetrathionate (NaTT) to induce disulfide crosslinks or at 37 degrees C to induce temperature-dependent insolubilization prior to extraction, transferred an additional 9-18% of the nuclear protein from the high salt extract to the nuclear matrix. This additional protein represented primarily an increased recovery of the same nuclear protein subset present in nuclear matrices prepared from untreated nuclei. Major constituents of chromatin including histones, hnRNP core proteins, and 98% of nuclear DNA were removed in the high salt extract following either incubation. Polyoma large T antigen was quantified in subcellular fractions by immunoblotting with rat anti-T ascites. Approximately 60 70% of the T antigen was retained in nuclei isolated in isotonic sucrose buffer at pH 7.2. Most (greater than 95%) of the T antigen retained in untreated nuclei was extracted by DNase-high salt treatment. Incubation at 37 degrees C or with NaTT transferred most (greater than 95%) of the T antigen to the nuclear matrix. T antigen solubilized from NaTT-treated matrices with 1% SDS sedimented on sucrose gradients as a large (50-S) complex. These complexes, isolated by immunoprecipitation with anti-T sera, were dissociated by reduction with 2 mercaptoethanol, and SDS-PAGE analysis revealed that T antigen was crosslinked in stoichiometric amounts to several host proteins: 150, 129, 72, and 70 kDa. These host proteins were not present in anti-T immunoprecipitates of solubilized nuclear matrices prepared from iodoacetamide-treated cells. Our results suggest that the majority of polyomavirus large T antigen in infected cells is localized to a specific subnuclear domain which is distinct from the bulk chromatin and is closely associated with the nuclear matrix. PMID- 3040448 TI - In vitro maturation of collagen fibrils modulates spreading, DNA synthesis, and collagenolysis of epidermal cells and fibroblasts. AB - Collagen fibrils were maturated in vitro by incubating them in a serum-containing culture medium at 37 degrees C for varied lengths of time. Epidermal cells and fibroblasts were cultured on these maturated collagen gels to see the effects of maturation on cellular morphology and physiology. The spreading and DNA synthesis of both types of cells on the maturated collagen gels were significantly enhanced compared to those on fresh gels. The maturation did not affect the cellular adhesiveness to the substrate. The secretion of collagenase by epidermal cells was suppressed on the maturated collagen gels, the extent of the suppression being related to the length of maturation of the gels. These maturation-related effects of collagen were also observed when collagen was incubated in the medium without serum, indicating that the effects are not due to deposition of serum proteins to collagen gels during maturation. Physical and chemical characterizations of the maturated collagen were performed: the mechanical strength of collagen gels increased in maturated collagen gels, the amounts of insoluble collagen increased with the maturation. These changes in the chemical and physical nature of the maturated collagen gel strongly suggested that there was an increase in intermolecular crosslinks during the process of maturation. These maturation-induced changes in collagen were marked when collagen gels were incubated in the presence of glucose, indicating that a glucose-protein reaction such as the Maillard reaction is involved in this phenomenon. PMID- 3040449 TI - TPA induces cytoplasmic alkalinization in human monoblastic U-937 cells without activation of Na+/H+ exchange. AB - The effect of the tumor-promoting phorbol ester 12-O-tetradecanoylphorbol-13 acetate (TPA) on cytoplasmic pH (pHi) and H+ extrusion was studied in the human monoblastic cell line U-937. About 2 min after addition of TPA, pHi started to increase and reached a steady state 10-15 min later. The resulting alkalinization corresponded to 0.03 and 0.09 pH units at 10(-10) and 10(-7) M TPA, respectively. The TPA-induced increase in pHi was independent of the presence of extracellular Na+. Moreover, TPA did not affect the H+ extrusion from the U-937 cells. Together these observations indicate the presence of a novel mechanism for TPA-induced cytoplasmic alkalinization. This mechanism is independent of Na+/H+ exchange across the plasma membrane, but may involve organelle sequestration of H+. PMID- 3040450 TI - Alpha-factor inhibition of the rate of cell passage through the "start" step of cell division in Saccharomyces cerevisiae yeast: estimation of the division delay per alpha-factor.receptor complex. AB - A highly sensitive, kinetically unambiguous assay for alpha-factor-induced delay of cell passage through the "start" step of cell division in yeast is presented. The assay employs perfusion with periodic microscopy to monitor the bud emergence kinetics on the 20% of cells within an exponentially growing population which exist prior to the alpha-factor execution point of start. The t1/2 for cell passage through start by this population of cells is 31 min in the absence of alpha-factor. The inhibition constant, KI, represents the alpha-factor concentration which produces a 50% inhibition of this rate and is equal to 2 X 10(-10) M. A second assay for maximal cell division arrest by alpha-factor on whole populations of cells is presented. This assay shows a maximum cell division arrest time of 125 +/- 5 h at saturating alpha-factor, and a K50 (that is, an alpha-factor concentration which produces a half-maximal response) of 2.5 X 10( 8) M. Both assays were performed in the effective absence of alpha-factor inactivation. Values of the dissociation constant KD and total number of receptors per cell which specifically mediate cell division arrest or delay were estimated to be 2.5 X 10(-8) M and 10(4), respectively. These estimates, along with the quantitative dose-response data for division arrest which are presented here, are consistent with each receptor.alpha-factor complex which is present on the cell at equilibrium producing a 43 +/- 10 s delay of cell passage through start. Surprisingly, this number is constant within twofold over the entire range of cellular division arrest responses to alpha-factor, that is, from a 1.9-fold inhibition of the rate of cell passage through start at 0.17 nM alpha-factor to a 125 +/- 5 h maximum arrest at saturating alpha-factor concentrations of greater than 170 nM. The possible significance of this observation toward the mechanism of alpha-factor-induced cell division arrest is discussed. PMID- 3040451 TI - Induction of cellular DNA synthesis in G0-specific ts mutant, tsJT60, following infection with SV40 and adenoviruses. AB - tsJT60 cells, a temperature-sensitive G0 mutant of a Fischer rat cell line, grew normally in an exponential growth phase at both permissive (34 degrees C) and nonpermissive (39.5 degrees C) temperatures, but when stimulated with fetal bovine serum in the growth-arrested state (G0 phase) they entered S phase at 34 degrees C but not at 39.5 degrees C. Infection of G0-arrested tsJT60 cells with SV40, adenovirus (Ad) 5 wild type and its E1B mutant dl313, and Ad12 wild type and its E1B mutants in205B, in205C, dl205, and in206B induced DNA synthesis at both temperatures. The DNA synthesized after virus infection was shown to be cellular by Hirt separation of DNA from SV40-infected cells and by CsCl equilibrium density gradient centrifugation of DNA from Ad5-infected cells. PMID- 3040452 TI - Use of electroporation for high-molecular-weight DNA-mediated gene transfer. AB - Electroporation was used to introduce high-molecular-weight DNA into murine hematopoietic cells and NIH3T3 cells. CCRF-CEM cells were stably transfected with SV2NEO plasmid and the genomic DNA from G-418-resistant clones (greater than 65 kb) was introduced into mouse bone marrow and NIH3T3 cells by electroporation. NEO sequences and expression were detected in the hematopoietic tissues of lethally irradiated mice, with 24% of individual spleen colonies expressing NEO. The frequency of genomic DNA transfer into NIH3T3 cells was 0.25 X 10(-3). Electroporation thus offers a powerful mode of gene transfer not only of cloned genes but also of high-molecular-weight DNA into cells. PMID- 3040454 TI - Fine-needle and screw-needle samples in CT-assisted biopsies of chest lesions. AB - We compared samples obtained using a fine-needle with those made using a screw needle (Rotex) of equal diameter in 28 patients. The biopsies were performed with the aid of computed tomography. All patients had lesions suggesting primary lung neoplasia or metastatic disease located in the lung parenchyma, pleura and in the ribs (17, 9 and 2 patients, respectively). Diagnostic material was obtained from 25 patients. There was no statistically significant difference in the yield of diagnostic material produced by the two procedures. Malignancy was found in 12 patients. Pneumothorax was the most common complication, occurring in 7 patients. CT-guided diagnostic needle biopsies have high diagnostic accuracy and a low complication rate. The diagnostic yield of the fine-needle and the screw-needle is equal. PMID- 3040453 TI - Influence of cytomegalovirus infection on the recovery of humoral immunity after autologous bone marrow transplantation. AB - Recovery of B-cell number and function was studied in 23 patients with hematological malignancies treated with high-dose chemoradiotherapy followed by autologous bone marrow transplantation (auto-BMT) in relation to the presence or absence of cytomegalovirus (CMV) infection. B cells recovered rapidly after auto BMT and specific antibodies to herpes viruses remained nearly unchanged. Both were independent of the CMV status of the patients. However, the capacity of peripheral blood B cells to differentiate in vitro into cytoplasmic immunoglobulin (Ig)-positive cells (plasma cells) on pokeweed mitogen stimulation in the presence of normal T-cell help was significantly better in CMV-negative patients than in CMV-positive patients after auto-BMT, but was decreased in both groups. Serum Ig levels were, in contrast, higher in CMV-positive patients than in CMV-negative patients after auto-BMT. PMID- 3040456 TI - Lateral excitation in the cat lateral geniculate nucleus. AB - Visual responses were elicited by global phase reversal stimuli in cells of the cat dorsal lateral geniculate nucleus (dLGN) after small retinal lesions had been centered on each receptive field. After acute lesions of different sizes exclusively lateral inhibition was found. When GABAergic inhibition was blocked by continuous microiontophoretic application of bicuculline lateral excitation emerged in dLGN cells partially deafferented by small and medium size acute retinal lesions, but not in those affected by large lesions. This indicates the presence of excitatory retinal inputs at the periphery of the dLGN cell dendrites which are normally suppressed by strong, long-ranging lateral inhibitory processes. After chronic deafferentation, the remaining excitatory inputs increase in effectiveness and lateral excitation is seen without blockade of inhibition. The maximal lateral spread of excitation (300 micron) in the dLGN is distinctly smaller than the extent of lateral inhibition (1000 micron). PMID- 3040455 TI - Dose-dependent influences on electrophysiological signs of attention in humans after neuropeptide ACTH 4-10. AB - The afferent humoral system exerts significant influences on brain activity. Central nervous actions of the adrenocorticotropic hormone (ACTH) are most likely to be mediated by information coded in a portion of this hormone structure corresponding to ACTH 4-10. Our previous research suggested an impairing effect of ACTH 4-10 on electrophysiological signs of selective attention in humans. The present experiments in 12 male subjects investigated the influences of ACTH 4-10 on different aspects of attention as indicated by auditory event-related potential (AERP) components. Furthermore, dose-response characteristics of these influences should be examined. Attention performance was tested in a dichotic listening paradigm, after 0, 0.1, 1.0, and 10 mg ACTH 4-10, administered intravenously 1 h prior to testing according to a double-blind latin-square design. Different aspects of attention were measured by brain electrical responses evoked either by frequent standard or rare target tone pips, which the subject had to attend to, or to ignore. The selective type of attention was reflected by the Nd determined as mean difference in amplitude between AERPs to tone pips when attended and when unattended, for a latency range between 0-460 ms post-stimulus. In addition, plasma cortisol, heart rate, blood pressure, and behavioral performance were measured. Results indicated a clear reduction of the Nd amplitude after all doses of ACTH 4-10. Other indicators of attention mechanisms such as mismatch processing were not affected by the peptide. The diminished Nd after ACTH 4-10 was due to an increased processing of unattended stimuli, but simultaneously attended tones were processed less intensively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3040457 TI - Excitatory inputs to cerebellar dentate nucleus neurons from the cerebral cortex in the cat. AB - 1. In anesthetized cats, we investigated excitatory and inhibitory inputs from the cerebral cortex to dentate nucleus neurons (DNNs) and determined the pathways responsible for mediating these inputs to DNNs. 2. Intracellular recordings were made from 201 DNNs whose locations were histologically determined. These neurons were identified as efferent DNNs by their antidromic responses to stimulation of the contralateral red nucleus (RN). Stimulation of the contralateral pericruciate cortex produced excitatory postsynaptic potentials (EPSPs) followed by long lasting inhibitory postsynaptic potentials (IPSPs) in DNNs. The most effective stimulating sites for inducing these responses were observed in the medial portion (area 6) and its adjacent middle portion (area 4) of the precruciate gyrus. Convergence of cerebral inputs from area 4 and area 6 to single DNNs was rare. 3. To determine the precerebellar nuclei responsible for mediation of the cerebral inputs to the dentate nucleus (DN), we examined the effects of stimulation of the pontine nucleus (PN), the nucleus reticularis tegmenti pontis (NRTP) and the inferior olive (IO). Systematic mapping was made in the NRTP and the PN to find effective low-threshold stimulating sites for evoking monosynaptic EPSPs in DNNs. Stimulation of either the PN or the NRTP produced monosynaptic EPSPs and polysynaptic IPSPs in DNNs. Using a conditioning-testing paradigm (a conditioning stimulus to the cerebral peduncle (CP) and a test stimulus to the PN or the NRTP) and intracellular recordings from DNNs, we tested cerebral effects on neurons in the PN and the NRTP making a monosynaptic connection with DNNs. Conditioning stimulation of the CP facilitated PN- and NRTP-induced monosynaptic EPSPs in DNNs. This spatial facilitation indicated that the excitatory inputs from the cerebral cortex to DNNs are at least partly relayed via the PN and the NRTP. 4. Stimulation of the contralateral IO produced monosynaptic EPSPs and polysynaptic IPSPs in DNNs. These monosynaptic EPSPs were facilitated by conditioning stimulation of the CP, strongly suggesting that the IO is partly responsible for mediating excitatory inputs from the cerebral cortex to the DN. A comparison was made between the latencies of IO-evoked IPSPs in DNNs and the latencies of IO-evoked complex spikes in Purkinje cells. Such a comparison indicated that the shortest-latency IPSPs evoked from the IO were not mediated via the Purkinje cells and suggested the pathway mediated by inhibitory interneurons in the DN.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3040458 TI - Acetylcholinesterase has a non-cholinergic neuromodulatory action in the guinea pig substantia nigra. AB - Acetylcholinesterase is released within the substantia nigra from the soma/dendrites of nigrostriatal neurons. Previous work suggests that this phenomenon is independent of cholinergic systems, but rather serves to modulate the sensitivity of dopamine-containing nigrostriatal cells to synaptic events. This hypothesis was tested directly in the anaesthetized guinea-pig. Micro infusion of acetylcholinesterase into the substantia nigra led to an increase in spontaneous firing of nigrostriatal neurons. Furthermore, the pattern of firing evoked by stimulation of the striatum was markedly enhanced. By contrast, administration of butyrylcholinesterase had no effect. It thus appears that acetylcholinesterase has a modulatory action on the firing of nigrostriatal cells, independent of hydrolysis of acetylcholine. PMID- 3040459 TI - Effects of denervation on calpain and calpastatin in hamster skeletal muscles. AB - Three leg muscles (soleus, extensor digitorum longus, and plantaris) of adult male golden Syrian hamsters were denervated by bilateral transecting of the sciatic nerve. Eighteen days after denervation, wet weights, amounts of soluble protein, and activities of wide-specificity calpain II, intermediate filament protein-specific calpain I, and calpastatin were measured by protein determination and enzyme and immunological assays. In comparison with control (nondenervated) muscles and depending on the specific muscle and protein, the activities of the calpains increased 1.3 to 1.9 times the control values, whereas the calpastatin decreased to one-half and one-third of control values. The muscle which showed the greatest increase in both calpain activities and the largest decrease in calpastatin activity was the plantaris (a fast-twitch, oxidative glycolytic muscle). The extensor digitorum longus (fast-twitch glycolytic) showed increases in calpain II activity similar to those in the plantaris, but smaller changes in calpain I and calpastatin. The soleus (slow-twitch, oxidative) showed the smallest changes in calpain II and calpastatin activities, although an increase in the calpain I activity was seen after denervation. These results suggest a possible relationship between the presence of fast-twitch, oxidative glycolytic fibers in a muscle and increased potential for intracellular proteolysis following denervation. PMID- 3040460 TI - Inhibition of heterosensory thalamocortical evoked potentials by delta-9 tetrahydrocannabinol. AB - The effects of delta-9-tetrahydrocannabinol on sensory activity in the thalamic intralaminar nuclei, centralis lateralis and the mesencephalic reticular formation were compared with the effects on cortical association or heterosensory systems in alpha-chloralose-anesthetized cats. The drug depressed the anterior marginal responses to multiple-modality sensory stimulation 30 min after administration of 2 mg/kg. Posterior suprasylvian responses were not significantly depressed except during 15 min postadministration. The drug did not depress sensory responses at the centralis lateralis or the mesencephalic reticular formation. Cortical responses evoked by stimulation of the latter were globally depressed by the drug. In contrast, delta-9-tetrahydrocannabinol depressed the anterior marginal response to centralis lateralis stimulation and not the posterior suprasylvian response similarly to the responses to sensory stimulation. The drug was without effect on caudate nucleus responses to stimulation of the mesencephalic reticular formation or centralis lateralis. These data demonstrate that, unlike pentobarbital, delta-9-tetrahydrocannabinol maintains heterosensory afferent activity to mesencephalic and thalamic sites and that its unique properties appear to be due, in part, to the selective disruption of heterosensory thalamocortical function. PMID- 3040461 TI - CCK-8 elicits grooming: cross tolerance to ACTH1-24. AB - Rats received lateral intracerebroventricular (icv) injection of sulfated or desulfated cholecystokinin octapeptide (CCK-8-S or CCK-8-D, respectively) and subsequent grooming behaviors were observed using a behavioral sampling technique. CCK-8-S (2.0 to 4.0 micrograms in 10 microliter) elicited a significant increase in grooming behaviors relative to the controls, but the relatively inactive CCK-8-D did not. Excessive grooming induced by CCK-8-S produced less grooming than did the adrenocorticotropic hormone fragment, ACTH1 24 (1.0 microgram in 10 microliter). An icv injection of CCK-8-S 3 h before the icv injection of ACTH1-24 inhibited the usual excessive grooming produced by the ACTH. Conversely, prior injection of ACTH1-24 abolished the increased grooming induced by CCK-8-S. Thus, the sulfated cholecystokinin octapeptide and ACTH1-24 exhibited short-term cross tolerance for excessive grooming. These results suggest that the colocalization of corticotropin releasing hormone and CCK-8 in neurons may have functional significance in connection with stress-related neuronal systems. PMID- 3040463 TI - Immunocytochemical localization of 5'-nucleotidase in myelinated peripheral nerves from the rat. AB - In sciatic and trigeminal nerves from the rat, 5'-nucleotidase immunostaining was observed on the surfaces of the myelinated fibers and in the membranes encircling the outermost loops of the myelin sheaths, the paranodal loops, and perhaps the inner loops, but neither in the compact myelin nor in the axoplasm. These results, which were consistent with previous biochemical data regarding sciatic nerve, suggest that the function of 5'-nucleotidase in myelinated fibers in the peripheral nervous system may be to promote diffusion of adenosine between the glial and neuronal compartments. PMID- 3040462 TI - Org.2766 protects from cisplatin-induced neurotoxicity in rats. AB - One of the side-effects of the cytotoxic drug, cisplatin, is its neurotoxicity. In rats this neurotoxicity can be measured as a slowing of the H-reflex-related sensory nerve conduction velocity. Concurrent treatment with Org.2766 (an ACTH4-9 analog) prevents this neurotoxic side effect while leaving the antitumor activity of cisplatin unaffected. PMID- 3040464 TI - The effect of cyclic cytidine 3',5'-monophosphate (cCMP) on the in vitro development, hatching and attachment of the mouse blastocyst. AB - The in vitro development and attachment of hatched mouse blastocysts on the untreated substratum was enhanced by 10 microM dibutyryl cCMP (dbcCMP). The result suggests that cCMP has an effect on embryonic development and on the blastocyst attachment process. PMID- 3040465 TI - Identification of free radicals in myocardial ischemia/reperfusion by spin trapping with nitrone DMPO. AB - The spin trapping ESR technique was applied to investigate oxygen-derived radicals in ischemic and post-ischemic rat hearts. Using 5,5'-dimethyl-l pyrroline-N-oxide, carbon-centered radicals were identified during ischemia and oxy-radical adducts (superoxide anion radical, O.-2 and hydroxyl radicals, .OH) in post-ischemic rat heart. The formation of these spin adducts was inhibited by superoxide dismutase, suggesting that superoxide plays a role in the adducts' formation. The results demonstrate that oxygen derived free radicals are important byproducts of abnormal oxidative metabolism during myocardial ischemic and reperfusion injuries. PMID- 3040467 TI - Rapid formation of secondary structure framework in protein folding studied by stopped-flow circular dichroism. AB - Kinetic refolding reactions of ferricytochrome c and beta-lactoglobulin have been studied by stopped-flow circular dichroism by monitoring rapid ellipticity changes of peptide backbone and side-chain chromophores. In both proteins, a transient intermediate accumulates within the dead time of stopped-flow mixing (18 ms), and the intermediate has an appreciable amount of secondary structure but possesses an unfolded tertiary structure. It is suggested that the rapid formation of a secondary structure framework in protein folding is a common property observed in a variety of globular proteins. PMID- 3040466 TI - Differential response of the human hepatoma-derived cell line HA22T/VGH to polypeptide mitogens. AB - Several human cell lines derived from primary cancer of the liver are able to grow under serum-free conditions and produce spreading and growth factors which are released into the culture medium. Since this autocrine growth under hormone free conditions might play a basic role in malignant transformation, we studied the effect on cell replication and the presence of specific membrane receptors of epidermal growth factor (EGF) and insulin on a dedifferentiated human hepatoma cell line, named HA22T/VGH. Our results point to a similar inhibitory effect on cell replication in the presence of both EGF and insulin, in spite of detecting different affinities of binding. PMID- 3040468 TI - Recombinant gamma-interferon stimulates iodide uptake and cyclic AMP production by the FTRL5 thyroid cell line. AB - The effect of recombinant rat gamma-interferon (gamma-IFN) on iodide uptake and cAMP production by rat thyroid cells in vitro was studied using the continuously growing, functional FRTL5 cell line. Both functions were stimulated by gamma-IFN at concentrations of 1-10 U/ml. Iodide uptake was dependent on protein synthesis, since it was blocked by cycloheximide treatment, but was not dependent on growth factors in calf serum routinely used for FRTL5 cell culture. These results show that gamma-IFN can stimulate thyroid cell function as well as aberrant Ia expression in vitro. PMID- 3040469 TI - Variations with position of replication errors due to exonuclease warm-up. AB - A.A mismatch errors occurring during poly(dA) replication with the Klenow fragment of E. coli DNA polymerase I have been quantified. The A/T ratio measured for chains extended by 1-25 nucleotides decreases by a factor of at least 15 from beginning to end. The deduced true error rate may decrease by a factor of 2.5 at each successive nucleotide addition. When ddATP is used instead of dATP, the ddA/T ratio indicates little variation of the misincorporation probability with position. Thus, the accuracy improvement in the first case is due to a warm-up of the proofreading function. PMID- 3040470 TI - The phosphorylation at Thr 124 of simian virus 40 large T antigen is crucial for its oligomerization. AB - SV40 large T antigen is phosphorylated at up to ten different amino acids clustered in an N-terminal and a C-terminal part of the polypeptide chain. The N terminal phosphorylated residues include Ser 123 and Thr 124. We have analyzed the oligomerization, the complex formation with the cellular oncoprotein p53 and the DNA-binding properties of T antigen from two different SV40 transformed cell lines which have either an amino acid exchange at Ser 123 to Phe (W7) or Thr 124 to Ile (D29). In comparison to wild-type T antigen both mutant T antigens have a slightly reduced binding affinity for both binding sites, I and II, of SV40 DNA. Phosphorylation at both residues of T antigen is not essential for formation of the complex with p53. Only the phosphorylation at Thr 124 seems to be critical for the formation of high molecular mass oligomers. Our data support the hypothesis that the oligomerization of T antigen seems to be implicated in viral DNA replication. PMID- 3040471 TI - 8-Bromoguanosine 3':5'-cyclic monophosphate decreases intracellular free calcium concentrations in cultured vascular smooth muscle cells from rat aorta. AB - The effects of 8-bromoguanosine 3':5'-cyclic monophosphate (8-Br cGMP) on intracellular free calcium concentrations ([Ca2+]i) in cultured rat aortic vascular smooth muscle cells (VSMCs) loaded with fura-2 were recorded microfluorometrically. Irrespective of whether VSMCs were at rest (in 5 mM K+ PSS), under Ca2+ depletion (in Ca2+-free medium for 10 min) and K+ depolarization (in high K+ PSS), [Ca2+]i was actively reduced and reached a new and lower steady state level with the application of 8-Br cGMP. This may be the first and direct evidence that cGMP, a putative mediator of various vasodilators, actively reduces [Ca2+]i in VSMCs. PMID- 3040472 TI - Methylation of the enhancer region of avian sarcoma virus long terminal repeat suppresses transcription. AB - The effect of methylation of an enhancer on transcription was studied. A 245 bp enhancer-containing a fragment of the LTR of the avian sarcoma virus was methylated in vitro and ligated back into a vector which lacked the upstream enhancer sequence. The transient expression in QT6 cells indicated that methylation of the enhancer-containing sequence severely reduced the extent of transcription. PMID- 3040473 TI - A stopped-flow study of the reaction of cytochrome c peroxidase with hydroperoxides. AB - Transient kinetic measurements show that cytochrome c peroxidase reacts with excess of hydroperoxides to produce compound ES in two phases. The activation energies for the fast and slow phases are calculated to be 6.3 and 20.5 kcal X mol-1, respectively. The fast phase is assigned to the reaction of native active (pulsed) cytochrome c peroxidase with peroxides, whereas the slow phase is due to the presence of an inactive (aged, resting) enzyme. As the active species is exhausted, the equilibrium between the active and inactive enzymes is shifted by a slow conformational change to replenish the active enzyme. Since the rate limiting step of the reaction of the inactive enzyme with peroxides is the conformation change, the overall reaction rate is independent of the nature and concentration of peroxides. PMID- 3040474 TI - Isolation and sequence analysis of a cDNA clone encoding the entire catalytic subunit of a type-2A protein phosphatase. AB - A 2.5 kb clone containing the full-length coding sequence of a type-2A protein phosphatase catalytic subunit has been isolated from a rabbit skeletal muscle cDNA library constructed in lambda gt10. The sequence of the protein deduced from the cDNA contains 309 residues (35.58 kDa). A major mRNA species at 2.0 kb and a minor component at 2.8 kb were visualized by Northern blotting in both skeletal muscle and liver. The type-2A enzyme showed weak homology with mammalian alkaline phosphatases between residues 55 and 95. The protein sequence of the type-2A phosphatase from rabbit skeletal muscle differs from that reported for the bovine adrenal enzyme in three regions. PMID- 3040475 TI - Oat cell carcinoma of the bronchus and acute pancreatitis. AB - A case of small cell carcinoma of the lung presenting with acute pancreatitis is described. We believe this to be the first such case reported, despite the frequency of metastasis to the pancreas from such tumours. We discuss the investigation and management of such cases. PMID- 3040476 TI - Use of single (4B-2) and repetitive copy (pS4) deoxyribonucleic acid (DNA) probes to characterize translocated Y DNA in a pedigree with recurrent abortion. AB - Probes for unique and repetitive copy deoxyribonucleic acid (DNA) are available to detect and characterize Y DNA. The probe pS4 detects repetitive copy DNA mapped to Yq12. The upper limits of the pS4 sequences are defined by the upper limits of C-banding. The probe 4B-2 is a recombinant phage construct developed from a Y library and contains a unique copy 3.3 kb Eco RI fragment mapped to Yq11. A family was ascertained through a pregnant female who had a history of four consecutive abortions and two normal daughters. Cytogenetic analysis revealed the mother and one of her daughters to have a 46,XX,15p+ karyotype. Amniocytes were karyotyped as 46,XY,15p+. Genomic DNA from controls, mother, daughters, and amniocytes was digested with Mbo I and hybridized to 32P-labeled pS4 probe. DNAs from both 46,XX,15p+ females and 46,XY,15p+ amniocytes demonstrated a clear male-specific 2.3 kb band. Digestion of the same genomic DNAs with Eco RI and blot hybridization to 32P-labeled 4B-2 probe revealed the 3.3 kb male-specific band only in the 46,XY,15p+ amniocyte DNA. The additional sequences located on 15p segregating in the female members of this family correspond to Yq12. The effect of this additional DNA on gametogenesis is unknown. PMID- 3040477 TI - [Age-related characteristics of the effect of acetylcholine, noradrenaline and vasopressin on the Na+, K+-ATPase activity of the plasma membranes of the nerve endings in the rat brain]. PMID- 3040478 TI - [Effect of cytochrome c antibodies on recovery processes in the liver and possible participation of anti-idiotypic antibodies in the realization of their action]]. PMID- 3040479 TI - [Participation of the nigro-striatal and mesolimbic dopaminergic systems of the brain in the control of components of learned motor reactions in the dog]. AB - Microinjections of dopamine (3 micrograms) into the caudate nucleus head and into the nucleus accumbens produced unidirectional effects on the motor components of avoidance behaviour and posture adjustment in two groups of the dogs (intact ones and dogs with different degree of caudate pathology). However, each of these dopaminergic systems can regulate mainly certain components of motor program, i.e. nigrostriatal system--the initiation of both the conditioned postural adjustment and the voluntary movement and mesolimbic system--the degree of this movement manifestation in amplitude and stability of realization. The greatest effect was shown after combined microinjection of dopamine bilaterally into both structures in the dogs with akinetic form of caudate pathology. The obtained data suggest that the integration of the striatal and limbic dopaminergic mechanisms seems to be critical for initiation and control of components of voluntary movement in the dog. PMID- 3040480 TI - [Direct effect of met-enkephalin on the synthesis and secretion of corticosterone by the adrenal glands of the rat]. AB - ACTH (1 unit per 1 rat) induced a 6-fold increase of the corticosterone synthesis in adrenals and a 2-fold increase of its secretion into the blood. Met-enkephalin (15 micrograms/kg) also increase the corticosterone secretion, the level of the hormone in the blood, however, becoming even lower. Combined action of these two substances involved a greater increase of the hormone concentration in the gland than in case of each of them separately. The hormone content in the blood remained low in this case, too. In perfusion of an isolated adrenal gland with met-enkephalin (15 micrograms/ml), no increase in the corticosterone concentration was observed either in the perfusate samples. The data obtained suggest that met-enkephalin activates synthesis of corticosteroids, on the one hand, and inhibits their secretion into the blood, on the other hand, whereas ACTH activates both these processes. PMID- 3040482 TI - Cytochemical and functional evaluation of ACTH responsive isolated rat adrenocortical cells. The maintenance of sex differences. AB - The critical evaluation of isolation methods for obtaining the adrenocortical cell suspension due to trypsin or collagenase digestion was done. Some collagenase advantages were indicated by morphological observations on the staining smears as well as by ACTH stimulation test. The cytochemical reactions for enzyme activities had the limited applications for those purposes. It also appeared that commonly applied dye exclusion tests were inadequate for characterization of cell suspension. The possible role of the adrenocortical cell debris in the basal corticosterone production was pointed out. The maintenance of the sex dimorphism and the functional differences in the adrenocortical cells isolated from male and female rats have been observed. PMID- 3040481 TI - [Effect of cholinergic substances on the spontaneous activity of lower respiratory tract neurons]. AB - Acetylcholine and low concentrations of nicotine were shown to increase firing rate, to induce grouping of spikes of spontaneously active cells and to activate "silent" neurons. High concentrations of nicotine exert a biphasic (excitatory inhibitory) effect. Atropine just alters and benzohexonium suppresses the spontaneous unit activity. Changes in the unit activity seem to stem from an interaction of cholinergic substances with the nicotine and muscarine cholinoreceptors. Neurons with a mixed type activity were found to receive inhibitory cholinergic projections. PMID- 3040483 TI - [Diurnal variation and responses of corticoids to dexamethasone and ACTH before and after adrenal surgery in primary aldosteronism]. AB - We studied diurnal variation and the responses of plasma corticoids to dexamethasone and ACTH before and after adrenal surgery in 11 patients with primary aldosteronism. Diurnal variation of plasma corticoids was examined in all cases. Before adrenal surgery, plasma aldosterone (Ald) was higher at all times, deoxycorticosterone (DOC) was high value at 5:00 and then normal value, 11 deoxycortisol (S) was high value at 5:00 and 9:00, thereafter normal value, while corticosterone (B) and cortisol (F) were almost normal value at all times. The circadian rhythm was observed in these corticoids. Dexamethasone (2 mg/day) was administered to 8 patients for 10 days. Plasma Ald and DOC were significantly suppressed only at 5:00, while B, S and F almost suppressed at all times. After dexa suppression, the circadian rhythm of Ald still observed, while the diurnal variation of the other corticoids was even. ACTH (1 microgram) was injected intravenously to 9 patients. Responses of DOC, B and Ald were higher, and of S and F were almost normal. Diurnal variation of plasma corticoids was observed after 1 month of adrenal surgery. Ald was lower, the other corticoids were normal values. ACTH (1 microgram) was injected to the same case before surgery. Responses of these corticoids to ACTH were slightly lower. Contents of Ald in adenoma tissues were higher than those in adjacent and normal adrenal tissues, and the contents of the other corticoids in adenoma tissue were almost normal value. As these results showed, before surgery these corticoids were secreted from adenoma without ACTH, although these were responsible to ACTH, and the presence of regulative factor of circadian rhythm except ACTH was suggested. After surgery, not only the responses to ACTH of mineralocorticoids but these of glucocorticoids decreased. PMID- 3040484 TI - [Bazex acrokeratosis in metastasizing bronchial cancer]. PMID- 3040485 TI - Thymic hypoplasia, splenomegaly and immune depression in guinea pigs with neonatal cytomegalovirus infection. AB - The effects of cytomegalovirus (CMV) infection on the spleen and thymus of neonatal guinea pigs were assessed. Guinea pigs with neonatally acquired CMV infection developed growth retardation, thymic hypoplasia and splenomegaly. Significant depletion of the T lymphocyte population occurred in the thymuses of these animals whereas inflammatory and immune proliferative responses were clearly evident in their spleens. Higher titers of infectious virus were recovered from the spleen than from the thymus. In addition, spleen cells from neonatally infected animals had significantly reduced proliferative responses to both the T-cell mitogen, concanavalin A, and the B-cell mitogen, lipopolysaccharide. Responses to concanavalin A were most severely impaired. These results point to the significant immunodepressive effect of acute CMV infection and to the dissimilar alterations induced by CMV in the spleen and thymus of acutely infected neonates. PMID- 3040486 TI - Effects of calcium ionophore A23187 and calcium antagonists on 32Pi incorporation into polyphosphoinositides of rat cortical synaptosomes. AB - The role of Ca2+ on 32Pi incorporation into polyphosphoinositides (PPI) of rat cortical synaptosomes was studied. Stimulation of muscarinic receptor by carbachol (1 mM) resulted in a decrease in 32Pi incorporation into phosphatidylinositol-4,5-bisphophaphate (TPI) and phosphatidylinositol-4 phosphate (DPI), and an increase in 32Pi incorporation into phosphatidylinositol (PI) and phosphatidic acid (PA), whereas no significant effect on other membrane phospholipids was found. This response could be blocked by atropine (1 microM). The stimulatory effect of carbachol required Ca2+ in the medium; the presence of 0.5 mM EGTA blocked the effect of carbachol on PPI turnover completely. Calcium ionophore A23187, at 1 microM, had a similar effect on PPI turnover by carbachol (1 mM). At higher concentrations (10-100 microM) of A23187, the PPI turnover rate was much enhanced. Depolarization of the membrane by high potassium (60 mM) in the presence of calcium resulted in an enhanced PPI turnover, which was similar to the results of the carbachol (1 mM) effect but to a lesser extent. Calcium antagonists, diltiazem and trifluoperazine, at 10 microM could block the carbachol effect on 32Pi incorporation into PPI in this preparation. Our results suggest that the enhancement of PPI turnover in rat cortical synaptosomes by carbachol, calcium ionophore or high potassium requires Ca2+, and it can be blocked by compounds which interfere with the availability of this ion, such as EGTA or calcium antagonists. PMID- 3040487 TI - Protein band 3 phosphotyrosyl phosphatase. Purification and characterization. AB - A phosphotyrosylprotein phosphatase has been purified from human red cell cytosol by successive DEAE cellulose, phosphocellulose and Red Procion-H3B-Sepharose chromatography. Overall purification was about 9000 with a yield of 30%. The enzyme was more than 95% pure as judged by SDS polyacrylamide gel. Its molecular weight was 17,000 and maximum activity was observed at pH 5.5. It was active towards both the phosphorylated tyrosine on the cytosolic fragment of the red cell protein band 3 and para-nitrophenyl phosphate. However the effects of ligands differ for the two substrates. PMID- 3040489 TI - Inositol phospholipids and cell regulation. PMID- 3040490 TI - Role of arachidonic acid metabolites in inflammatory and thrombotic responses. PMID- 3040488 TI - Effect of ligands on Drosophila phosphorylase a as monitored by its enzymic inactivation. AB - The dephosphorylation of Drosophila phosphorylase a with the catalytic subunit of fruit-fly protein phosphatase-1 was inhibited by AMP, IMP, ADP, ATP, glucose-6-P, glucose-1-P and UDPG. Glucose, caffeine and glycogen did not influence the reaction. The inhibitory effect of AMP was reduced by glucose and caffeine. The above ligands acted through the modification of phosphorylase a conformation. This conclusion was drawn from the ligands' effect on the dephosphorylation of phosphohistone by Drosophila phosphatase-1 and on the tryptic digestion of fruit fly phosphorylase a. PMID- 3040491 TI - Regulation of cholesterol metabolism in the liver in vivo and in vitro. PMID- 3040492 TI - Oxygen gradients: the problem of hypoxia. PMID- 3040493 TI - Peritubular cells influence Sertoli cells at the level of translation. AB - Conditioned medium from cultured peritubular cells (PTCM) was capable of increasing the incorporation of amino acids into acid-precipitable material in cultured Sertoli cells, while the incorporation of uridine into acid-precipitable material was unaffected. PTCM did not influence intracellular cAMP accumulation in a manner similar to follicle-stimulating hormone (FSH). PTCM was able to stimulate androgen-binding protein (ABP) secretion by Sertoli cells even in the presence of a maximal dose of FSH. PTCM increased the rate at which peptides are elongated 5-fold over control medium or medium from control fibroblasts. These studies indicate that peritubular cells influence Sertoli cells through different mechanisms than FSH and exert their influence, at least in part, at the level of translation by increasing the rate of peptide elongation. PMID- 3040494 TI - Modulation of thyroglobulin release by dog thyroid slices. AB - Nonbutanol-extractable 131I (NBE131I) release by dog thyroid slices in vitro has been shown previously to be primarily thyroglobulin (Tg); it is stimulated by TSH. NBE131I (Tg) release has therefore been considered as an in vitro model of thyroglobulin secretion and was further characterized in this work. TSH stimulated NBE131I (Tg) release, like TSH-stimulated BE131I (T4, T3 and iodide) release was reproduced by forskolin, an activator of adenylate cyclase. TSH-, (Bu)2cAMP- and forskolin-stimulated NBE131I (Tg) release was inhibited by 10(-5) M carbamylcholine, an effect relieved by 10(-5) M atropine, but not by 10(-4) M 1 methyl-3-isobutylxanthine. NBE131I (Tg) release was observed in the presence of 2 mM methimazole and 2 mM perchlorate. Cooling the slices to 20 degrees C or addition of 10(-5) M monensin completely blocked the formation of apical pseudopods and BE131I release but not NBE131I (Tg) release. Inhibition by 500 microM chloroquine of intralysosomal Tg hydrolysis and BE131I release did not enhance NBE131I (Tg) release. Cytochalasin B induced a concentration-dependent increase in basal and TSH-stimulated NBE131I (Tg) release at concentrations which depressed TSH-stimulated BE131I release. Removal of Ca2+ from the medium and slices by 10(-3) M or 10(-4) M EGTA increased NBE131I (Tg) release. In conclusion, in dog thyroid slices, TSH-stimulated NBE131I (Tg) release was mediated by cAMP and inhibited by 10(-5) M carbamylcholine at a step beyond cAMP. It was not neosynthesized Tg. It did not seem to require the formation of apical pseudopods or to result from the escape from lysosomes of undegraded thyroglobulin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3040495 TI - cAMP dependent and independent regulation of thyroglobulin synthesis by two clones of the OVNIS 6H thyroid cell line. AB - The hormonal regulation of thyroglobulin synthesis has been studied using two independent clones of the OVNIS 6H cell line. Insulin, hydrocortisone and TSH were able to stimulate thyroglobulin synthesis, whereas transferrin, somatostatin and glycyl-histidyl-lysine were without effect. Insulin stimulated thyroglobulin synthesis without affecting cAMP production. Hydrocortisone, when combined with insulin was a stimulator too; this stimulation was not accompanied by an increase in cAMP. TSH alone was unable to stimulate either cAMP or thyroglobulin synthesis. The stimulatory effect of TSH on thyroglobulin synthesis took place only when combined with insulin or insulin plus hydrocortisone, and was mediated by cAMP. Consequently, insulin and hydrocortisone stimulated thyroglobulin synthesis by cAMP-independent mechanisms, whereas TSH acted via the cAMP system. Forskolin mimicked TSH effects on cAMP and thyroglobulin synthesis. Calf serum inhibited cAMP and thyroglobulin production. Optimal cAMP and thyroglobulin synthesis as well as TSH responsiveness were obtained in serum-free medium supplemented with 5 micrograms/ml insulin, 100 nM hydrocortisone and 1 mU/ml TSH. PMID- 3040496 TI - Interferon-gamma: pleiotropic effects on a rat pancreatic beta cell line. AB - We have recently shown that interferon-gamma (IFN-gamma) markedly upregulates the expression of the class I major histocompatibility proteins on pancreatic beta cells and have therefore postulated that interferon-gamma may enhance cytotoxic lymphocyte-mediated beta cell damage in insulin-dependent diabetes mellitus. To further explore the interaction between interferon-gamma and the pancreatic beta cell we have used the RIN-m5F insulinoma line to define the effects of interferon gamma on major histocompatibility protein expression, (pro)insulin and protein synthesis and cell growth. Interferon-gamma induced a dose-dependent increase in the expression of the class I major histocompatibility proteins on the RIN-m5F cells, the maximal increase (10-fold) being seen at an interferon-gamma concentration of 1 U/ml. The induction of class I proteins by interferon-gamma was nearly completely abolished by cycloheximide. Expression of class II (Ia) proteins was not detected either in the presence or absence of interferon-gamma. (Pro)insulin and protein synthesis were decreased by 60% and 40%, respectively, in RIN-m5F cells cultured with interferon-gamma (10 U/ml). Furthermore, the growth of RIN-m5F cells was significantly inhibited, and corresponding changes in cell morphology were evident, after 3 days of exposure to interferon-gamma (10 U/ml). These findings indicate that, in addition to its potential role in amplifying cytotoxic T cell activity against the pancreatic beta cell, IFN-gamma may also directly inhibit beta cell function and growth. Several mechanisms could therefore account for an ability of IFN-gamma to compromise beta cell function and contribute to the pathogenesis of insulin-dependent diabetes. PMID- 3040497 TI - Uptake and retention of moulting hormones by the integument of crayfish in vitro. III. The possible involvement of Na+/K+-ATPase. AB - The specific uptake of the ecdysteroid [3H]ponasterone A into crayfish hypodermis in vitro is inhibited by increasing concentrations of potassium in the medium. The Na+/K+-ATPase blocker ouabain also decreases uptake of the ecdysteroid. Maximal inhibition of uptake is reached at 5 X 10(-5) M ouabain. PMID- 3040498 TI - Binding of vasoactive intestinal polypeptide to dispersed enterocytes results in rapid removal of the NH2-terminal histidyl residue. AB - Specific binding sites for 125I-labelled vasoactive intestinal polypeptide (VIP) (half-maximal inhibition at 1.5 +/- 0.2 nM VIP) were identified on dispersed porcine enterocytes. Radioactivity bound to the cell surface was internalized. At 37 degrees C, a steady state was achieved after 45 min with a ratio of internalized to cell surface-bound radioactivity of approximately 1:2 but at 10 degrees C, no radioactivity appeared intracellularly. Incubation of VIP with cells in the absence of inhibitors of proteolysis for as short a time as 30 s at 37 degrees C led to the formation of [des-His1]VIP by the action of amastatin- and bestatin-sensitive aminopeptidase(s). This metabolite was formed in the presence of sodium azide and when incubations were performed at 10 degrees C suggesting that internalization was not a prerequisite for degradation. As [des His1]-VIP has only 1% of the bioactivity of VIP, formation of this metabolite will effectivily terminate the action of VIP in the epithelial layer of the intestine. PMID- 3040499 TI - Three translationally regulated mRNAs are stored in the cytoplasm of clam oocytes. AB - In situ hybridization was used to examine the spatial distributions of three translationally controlled maternal RNAs in oocytes and two-cell embryos of the clam Spisula. 3H-labeled single-stranded RNA probes were generated from SP6 recombinant clones containing DNA inserts encoding portions of histone H3 (the DNA sequence which is presented here), cyclin A, and the small subunit of ribonucleotide reductase. Hybridization of these probes to oocytes, in which the mRNAs are translationally inactive, shows that these mRNAs are stored in the cytoplasm. There is no evidence for sequestration of any of the RNAs within the nucleus or any other discrete structure. Instead they appear to be evenly distributed throughout the cytoplasm. PMID- 3040500 TI - Virus-associated receptors for polymerized human serum albumin (RpHSA) in patients with chronic active hepatitis B treated with recombinant leukocyte A interferon. AB - Hepatitis B surface antigen (HBsAg)-associated receptors for polymerized human serum albumin (RpHSA) are assumed to mediate viral attachment to hepatocytes in hepatitis B virus (HBV) infection. RpHSA was found to be coded by the pre-S region of HBV genome. Recently, the antiviral effect of recombinant leukocyte alpha-interferon was shown in patients with hepatitis B. Our study evaluated the detection and the clinical implications of RpHSA in patients with chronic active hepatitis B under treatment with recombinant alpha-interferon. Two out of nine patients eliminated all HBV markers including RpHSA. Four out of nine patients became negative for markers of viral replication but remained positive for HBsAg and in part for RpHSA. In three out of nine patients HBV markers including RpHSA remained unchanged. In summary, the titer for RpHSA is a reliable indirect marker for infectivity and of prognostic value in patients with chronic active hepatitis B during interferon treatment. Future studies should evaluate a putative immune response to RpHSA-containing viral surface proteins, which could be relevant for viral clearance in HBV infection. PMID- 3040501 TI - Investigations of amitraz neurotoxicity in rats. II. Effects on visual evoked potentials. AB - As a part of a series of studies investigating the possible neurotoxicity of amitraz (AMZ), a formamidine pesticide, visual evoked potentials were recorded from Long-Evans rats following acute and short-term repeated exposures to AMZ. The first of three experiments examined the relationship between a single ip injection of AMZ (0, 50, and 100 mg/kg) and the latency and peak-to-peak amplitude of pattern-reversal (PREP) and flash-evoked potentials (FEP). The effects of another formamidine, chlordimeform (CDM; 40 mg/kg), were also studied for comparison purposes. Two hours after treatment, AMZ exposure produced large, dose-related increases in PREP amplitudes. Exposure to CDM produced similar changes. Neither compound changed FEP amplitudes. Body temperatures were reduced and evoked potential peak latencies were increased by both compounds. The latency increases were probably a secondary consequence of hypothermia. In the second experiment, PREPs were recorded before and 2, 24, 48, and 72 hr after treatment with AMZ (100 mg/kg). The time course of changes was biphasic in nature, with increases in amplitudes (N1P1, P1N2, and N2P3) 2 hr after treatment followed by subsequent depression in amplitude (P2N3) at 48 hr. Recovery occurred by 72 hr after treatment. The third experiment examined the effects of three daily treatments with either vehicle or 50 or 100 mg/kg AMZ. Body weights and body temperatures showed dose-related reductions which progressed with each additional treatment and recovered partially by 6 days after cessation of treatment. The PREPs of AMZ-treated rats agains showed biphasic changes, with N1P1 and P1N2 amplitudes significantly increased on each day of treatment and 1-2 days following the third treatment. Amplitude P2N3 showed an initial increase on the first 2 days of treatment, followed by subsequent, progressive amplitude reductions. In summary, AMZ produced two phases of change in visual evoked potentials. The first phase was characterized by large increases in PREP amplitudes without increasing FEP amplitudes in the same rats. The second phase was characterized by suppression of PREP P2N3 amplitude. Short-term repeated exposure produced signs of accumulating intoxication including progressive loss of body weight, lowered body temperature, and prolonged duration of evoked potential changes. PMID- 3040502 TI - Evaluation of host resistance and immunity in mice exposed to the carbamate pesticide aldicarb. AB - Adult female Swiss-Webster and B6C3F1 mice received distilled water only or water containing 0.1, 1.0, 10, 100, or 1000 ppb of aldicarb daily for 34 days. The target concentration of aldicarb present in the 10 ppb dosing solution was analytically verified on a daily basis as was its stability over a 48-hr period. To develop an immune profile of this compound, functional parameters measured after exposure included resistance to infectious viral challenge; quantitation of splenic antibody-forming cells to sheep erythrocytes and circulating serum antibody levels; splenic lymphocyte blastogenesis to T- and B-cell mitogens; and mixed-lymphocyte culture response. To supplement the functional assays, complete blood counts, differential leukocyte counts, and body and relative organ weights were measured. In addition, gross and histopathologic examinations of tissues relevant to the immune system were performed. The absence of significant effects on any of these parameters suggests that aldicarb at environmentally relevant exposure concentrations is not immunotoxic in rodents. PMID- 3040503 TI - [Adrenergic receptors in physiopathology and clinical medicine: current role and research prospectives]. PMID- 3040504 TI - Protein kinase C in the regulation of smooth muscle contraction. AB - The cellular and molecular mechanisms underlying smooth muscle contraction are reviewed in the light of recent studies of smooth muscle ultrastructure and of the role of polyphosphoinositide turnover and protein kinase C function in smooth muscle contraction. A new model of smooth muscle contraction is proposed that differs radically from accepted views, particularly the latch bridge hypothesis, in terms of both Ca2+ messenger function and the molecular events underlying this process. A coordinate fibrillar domain model of contraction is proposed in which the initial and sustained phases of contraction are mediated by different cellular and molecular events. The initial phase of response is mediated by a rise in [Ca2+]c and the resulting calmodulin-dependent activation of both myosin light chain kinase and the dissociation of caldesmon from the actin-caldesmon tropomyosin-myosin fibrillar domain. These events lead to an interaction between actin and the phosphorylated light chains of myosin just as in previous models. However, this initial phase is followed by a sustained phase in which a rise in [Ca2+]sm stimulates the plasma membrane-associated, Ca2+-sensitive form of protein kinase C that results in the phosphorylation of both structural and regulatory components of the filamin-actin-desmin fibrillar domain. These events underlie the tonic phase of contraction. PMID- 3040505 TI - Leukotrienes in health and disease. AB - The leukotrienes (LTs) are 5-lipoxygenase metabolites of arachidonic acid. The synthesis and release of LTs have been demonstrated in many cells and organs, and LTs are considered to be normal products of continuous metabolism of arachidonic acid. However, although evidence in favor of a critical role for LTs in regulation of physiological functions is still scarce, a growing body of evidence suggests a role for LTs in mediation of several pathophysiological processes such as generalized or local immune reactions, inflammation, asthma, shock, and trauma. LTs have been shown to have potent actions on many essential organs and systems, including the cardiovascular system (heart, blood vessels, microcirculation), the pulmonary system (lung, airways), the central nervous system (neural, glial, and vascular elements), the gastrointestinal tract, and the immune system. In these organs the effects of LTs are mediated by specific LT receptors. Identification of LTs and characterization of their regional and systemic pathological effects, together with characterization of their receptors and elucidation of their structure-activity relationships, are fundamental to developing LT antagonists or synthesis inhibitors that might prevent or reverse LT-dependent reactions. Preliminary reports have already shown that such pharmacological agents ameliorate some aspects of disease processes in experimental animals as well as in humans. In this brief review we intend to highlight the evidence that implicates LTs in normal physiological functions as well as in disease processes. PMID- 3040506 TI - Percutaneous creation of a hepaticojejunostomy. AB - A procedure is described whereby a direct anastomosis may be created percutaneously between the biliary system and a Roux-en-Y limb; in effect a percutaneous hepaticojejunostomy. Although it will have limited applications, it represents a further extension of percutaneous techniques in the biliary system for complex problems. PMID- 3040507 TI - Cowden's disease: a case report and literature review. AB - Cowden's disease, or multiple hamartoma syndrome, is an uncommon condition with characteristic mucocutaneous lesions associated with abnormalities of the breast, thyroid, and gastrointestinal tract. We describe a 51-year-old man with hyperplastic polyposis of the entire alimentary tract as the most prominent feature of this disease. We also present a review of 85 cases of this entity as reported in the English medical literature, and summarize the pertinent findings. PMID- 3040508 TI - A study on the cell kinetics of the canine gastric mucosa by the cytofluorometric method: an evaluation of chemically induced gastric cancer. AB - The cell kinetic alteration in the background mucosa of canine gastric cancer induced by N-ethyl-N'-nitro-N-nitrosoguanidine (ENNG) was evaluated by cytofluorometry in which the rate of S and G2 + M phase cell in gastric mucosal cells could be calculated, and a triphasic alteration was demonstrated; an initial reduction phase, an increase phase and a plateau phase with a high value. The initial reduction phase was caused by non-specific toxicity of ENNG as observed in drug induced gastric mucosal lesions, and subsequent increase and plateau phases originated from the action of ENNG itself to activate the mucosal turn-over and from histological changes in the background mucosa such as regenerative hyperplastic change after mucosal erosion and atrophic changes, sometimes including intestinal metaplastic change. Further, in comparison to carcinogenesis in chemically induced gastric cancer with and without a surfactant (Tween 60), it was suggested that one of the promotion effects of Tween 60 was closely related with activation of the mucosal turn-over. PMID- 3040510 TI - Estrogen receptors in hepatocellular carcinoma: is endocrine therapy for hepatocellular carcinoma likely to be effective? AB - Specimens of human liver obtained at the time of operation were assayed for cytosolic estrogen receptors by the binding assay and enzyme immunoassay (EIA). Mean estrogen receptor contents determined by the binding assay were 17.8 fmol/mg protein in non-cirrhotic liver, 7.1 in cirrhotic liver, and 0.7 in hepatocellular carcinoma, by EIA the contents were 12.1, 5.9, and 0.8 fmol/mg protein, respectively. There were significant differences among the three groups. In particular, hepatocellular carcinoma specimens contained very little or no detectable amounts of estrogen receptors in either assay. The correlation between the estrogen receptor content determined by the binding assay and that determined by EIA was significant (r = 0.822, p less than 0.001). It is suggested that the estrogen receptor content decreases with the development of liver cirrhosis and hepatocellular carcinoma and that antiestrogen endocrine therapy for hepatocellular carcinoma may be ineffective. PMID- 3040511 TI - Ultrastructural localization of type IV collagen, laminin and prolyl hydroxylase in biliary epithelial cells of rat liver following ligation of the common bile duct. AB - Type IV collagen and laminin are major components of basement membrane (BM), whereas prolyl hydroxylase (PH) is a key enzyme in the hydroxylation of proline to hydroxyproline in collagen synthesis. In order to elucidate the exact mechanism of the formation of BM, immune electron microscopic observation of type IV collagen, laminin and PH was made in rat liver with marked proliferation of bile ducts following ligation of the common bile duct. Extracellular localization of type IV collagen was found in the BM of bile ducts and blood vessels and in the space of Disse in both normal rat liver and the liver of rats undergoing operation. Type IV collagen was localized in lamina rara and lamina densa. Laminin was codistributed with type IV collagen in BM but rarely in the space of Disse even in the liver of rats undergoing operation. Immunostaining of laminin was diffusely spread in lamina densa, but sparsely in lamina rara. Though no reaction products of type IV collagen and laminin were detected in the cytoplasm of normal biliary epithelial cells, they were found in rough endoplasmic reticulum (rER) and the vesicles close to the basal surfaces of the plasma membrane of the proliferating biliary epithelial cells. No evident localization of these components in Golgi apparatus was found. PH was found in rER of the biliary epithelial cells, hepatocytes, endothelial cells of vessels, fibroblasts and perisinusoidal cells except for Kupffer cells in normal rat liver. More intense and diffuse staining of PH was observed in rER in the proliferating biliary epithelial cells of the liver of rats undergoing operation in concomitance with the evident localization of type IV collagen in this organelle. These findings suggest that the major components of BM, such as type IV collagen and laminin in the proliferating biliary epithelial cells, are produced in rER and secreted by vesicles to the basal extracellular spaces, thus forming new BM in these circumstances. PMID- 3040509 TI - Natural killer activity in patients with chronic hepatitis treated with OK432, interferon, adenine arabinoside and glycyrrhizin. AB - Natural killer (NK) activity in the peripheral blood of patients with chronic liver disease was measured using 51Cr labeled K562 cells as target cells. NK activity was elevated but not significantly in patients with chronic hepatitis compared with healthy controls and significantly lower in the patients with hepatocellular carcinoma. The activity decreased in the order of chronic hepatitis, liver cirrhosis and hepatocellular carcinoma. Although the level of NK activity in patients with chronic hepatitis did not correlate with the level of alanine aminotransferase (ALT), it tended to be elevated in association with elevation of ALT in patients treated with OK432, interferon-beta, glycyrrhizin or adenine arabinoside. In chronic liver disease, phytohemagglutinin (PHA) skin test showed a positive correlation with NK activity. In all patients who were treated with the immunopotentiator, OK432, and whose HBeAg became negative, NK activity was elevated during the treatment. These results suggest that the NK activity in peripheral blood may be related to hepatocytic injury even if this is not the effector mechanism of the injury. PMID- 3040512 TI - Efficacy of commercial condoms in the prevention of hepatitis B virus infection. AB - Tips of synthetic and natural condoms were filled with serum samples containing either hepatitis B virus, herpes simplex virus, or cytomegalovirus, then fit over an 8-in. mechanical vibrator and inserted vibrating into sterile bath solutions for 30 min. Phosphorus 32-labeled hepatitis B and cytomegalovirus molecular probes and viral culture techniques for herpes simplex and cytomegalovirus were used to determine whether leakage of virus had occurred into the surrounding bath solutions. Natural condoms allowed leakage of hepatitis B virus but not herpes simplex virus or cytomegalovirus, whereas synthetic condoms prevented leakage of all viruses. These results suggest that natural condoms might not be effective in preventing sexually transmitted hepatitis B virus infection. PMID- 3040513 TI - Neuropathic intestinal pseudoobstruction. PMID- 3040515 TI - Effects of an airabrasive polishing system on restored surfaces. PMID- 3040514 TI - [The menstrual cycle after hormonal growth inhibition in girls]. AB - Up to 1979 growth was inhibited with hormones in 43 girls in puberty with constitutional gigantism. The average reduction in size as compared to the calculated final body height was 7.5 cm, treatment having been started at a skeletal age of 12.3 years and with a treatment duration of 1.4 years. In over 90% of the cases, the first spontaneous menstruation after termination of therapy occurred within three months. During the first two years, cycles were likewise regular in 90% of the cases, with a duration of 4 +/- 1 weeks. Ovarian function in adolescence was thus found not to be impaired, but rather to be more stable as compared to the total population. It may be concluded from this that the preconditions for future fertility can be regarded as favorable. PMID- 3040516 TI - An unusual case of dental erosion caused by nitric acid. PMID- 3040517 TI - Diuretic action and immunological cross-reactivity of corticotropin and locust diuretic hormone. AB - A functional similarity and immunological cross-reactivity between adrenocorticotrophic hormone (ACTH) and a locust diuretic hormone (DH) is reported. The functional similarity is expressed in that ACTH mimics DH by stimulating fluid secretion and cyclic AMP (cAMP) secretion in locust Malpighian tubules. Desacetyl-alpha-melanocyte-stimulating hormone is active to a lesser degree but no other POMC-derived peptide tested was found to follow suit. Immunological cross-reactivity is shown by a positive response of HPLC-purified DH with a specific ACTH radioimmunoassay as well as a significant reduction in DH activity (fluid secretion) after incubations with ACTH antiserum. However, ACTH and DH are different peptides since they do not share common separation characteristics on HPLC and ACTH does not induce a high excess secretion of cAMP by the tubule cell as does DH. PMID- 3040518 TI - Effects of cortisol and growth hormone replacement on osmoregulation in hypophysectomized coho salmon (Oncorhynchus kisutch). AB - Both cortisol and GH were able to reverse partially the effects of hypophysectomy on coho salmon (Oncorhynchus kisutch) as judged by improved seawater (SW) tolerance after long-term treatment; however, neither hormone significantly restored gill Na+, Na+,K+-ATPase activity. In the first experiment, gill Na+,K+ ATPase activity remained low in hypophysectomized (Hx) coho treated with cortisol (15 micrograms/g body wt, suspended in cocoa butter) for 96 hr (48 hr in fresh water followed by 48 hr in seawater). In addition, plasma sodium levels were higher in the cortisol-treated Hx animals compared with those in untreated controls. In the second experiment, treatment with cortisol-filled Silastic capsules and implants of bovine GH (mixed with cholesterol in a ratio calculated to yield a dose of 1.5 microgram/g body wt/week) for 12 days also failed to alter gill Na+,K+-ATPase activity, but did reduce plasma sodium levels in animals transferred to SW for 48 hr. These results suggest that these hormones may be involved in the development of hypoosmoregulatory ability in coho salmon. However, the failure of either hormone to restore gill Na+,K+-ATPase activity suggests that a combination of these hormones and/or an additional hormone(s) acting in a synchronized fashion may be required for full effectiveness. PMID- 3040519 TI - Hormonal regulation of hepatic glycogenolysis in the toad, Xenopus laevis, is mediated by cyclic AMP and not Ca2+. AB - Hepatic glycogenolysis and glycogen phosphorylase a activity were stimulated by arginine vasotocin (AVT) in liver pieces from Xenopus laevis cultured in vitro. In each case, the EC50 was about l nM. The increased rate of glycogenolysis brought about by either AVT or adrenaline was maintained for at least 6 hr and was unchanged when Ca2+ salts were omitted from the medium or when 2.5 mM EGTA was added. Neither the Ca2+ ionophore, A23187, nor the Ca2+ channel blocker, verapamil, had any effect on the rate of glycogenolysis in the presence or the absence of either hormone. Tissue cyclic AMP levels were unchanged by addition of AVT alone but were doubled in the presence of AVT plus either of the phosphodiesterase inhibitors, isobutylmethylxanthine or RO20-1724. These findings suggest that hormones regulating hepatic glycogenolysis in X. laevis use cyclic AMP, and not Ca2+, as an intracellular messenger. We would argue that cytosolic Ca2+ may not have become involved in regulation of hepatic glycogenolysis until after the ancestors of present day amphibians separated from those of present day mammals. PMID- 3040520 TI - Thermodynamic analysis of rat brain opioid mu-receptor-ligand interaction. AB - Opioid mu-receptors are membrane bound receptors. The mechanism by which they transduce their biological effect into the inner compartment of the postsynaptic cell is still not fully understood. The present study was attempted to the measurement of changes of the thermodynamic parameters of the receptor- agonist/antagonist interaction. We have set up the binding assays of a mu receptor agonist (3H-dihydromorphine) as well as an antagonist (3H-naloxone). The saturation isotherms of both ligands have been assayed at various temperatures and from the resulting KD values the standard changes of Gibbs energy, enthalpy and entropy have been calculated. While the binding of the mu-receptor agonist 3H dihydromorphine appears to be entropy driven (delta S0 = 230 J mol-1 K-1) and endothermic (delta H0 = 19 kJ mol-1), the binding of the mu-receptor antagonist 3H-naloxone is apparently driven by a decrease of standard enthalpy (delta H0 = 27 kJ mol-1; i.e. the reaction is exothermic) and is also characterized by an increase of standard entropy (delta S0 = 76 J mol-1 K-1). The maximal number of 3H-naloxone binding sites has to be determined by incubation at 0-4 degrees C. The present data to not support the view that opioid mu-receptors transduce their biological signal through the adenylatecyclase system by a mechanism similar to beta-adrenergically stimulated adenylatecyclase. PMID- 3040521 TI - Effect of a single neonatal treatment with steroid hormone or steroid-like molecules on myocardial ouabain binding in the adult rat. AB - A single neonatal treatment of rats with vitamin D3, gibberellin, allylestrenol or diethylstilbestrol (DES) influenced the ouabain binding capacity of myocardial Na, K-dependent ATP-ase. Of the active molecules tested, vitamin D3, DES and gibberellin had appreciable impact on myocardial ouabain receptors, enhancing and depressing their activity, respectively. The thymic dexamethasone and uterine estrogen receptors did not alter their binding capacity in response to neonatal exposure to vitamin D3 or gibberellin. PMID- 3040522 TI - Changes in vesicular membrane ESR spin label parameters upon isotope solvent substitution. PMID- 3040523 TI - The cloning and mapping of ADR6, a gene required for sporulation and for expression of the alcohol dehydrogenase II isozyme from Saccharomyces cerevisiae. AB - The alcohol dehydrogenase II (ADH2) gene of the yeast, Saccharomyces cerevisiae, is not transcribed during growth on fermentable carbon sources such as glucose. Growth of yeast cells in a medium containing only nonfermentable carbon sources leads to a marked increase or derepression of ADH2 expression. The recessive mutation, adr6-1, leads to an inability to fully derepress ADH2 expression and to an inability to sporulate. The ADR6 gene product appears to act directly or indirectly on ADH2 sequences 3' to or including the presumptive TATAA box. The upstream activating sequence (UAS) located 5' to the TATAA box is not required for the Adr6- phenotype. Here, we describe the isolation of a recombinant plasmid containing the wild-type ADR6 gene. ADR6 codes for a 4.4-kb RNA which is present during growth both on glucose and on nonfermentable carbon sources. Disruption of the ADR6 transcription unit led to viable cells with decreased ADHII activity and an inability to sporulate. This indicates that both phenotypes result from mutations within a single gene and that the adr6-1 allele was representative of mutations at this locus. The ADR6 gene mapped to the left arm of chromosome XVI at a site 18 centimorgans from the centromere. PMID- 3040524 TI - Analysis of the promoter of the ninaE opsin gene in Drosophila melanogaster. AB - We have analyzed the cis-acting regulatory sequences of the ninaE gene. This gene encodes the major Drosophila melanogaster opsin, the protein component of the primary chromophore of photo-receptor cells R1-R6 of the adult eye. DNA fragments containing the start point of transcription of the ninaE gene were fused to either the Escherichia coli chloramphenicol acetyltransferase or lacZ (beta galactosidase) gene and introduced into the Drosophila germline by P-element mediated transformation. Expression of the E. coli genes was then used to assay the ability of various sequences from the ninaE gene to confer the ninaE pattern of expression. Fragments containing between 2.8 kb and 215 bp of the sequences upstream of the start of transcription plus the first 67 bp of the untranslated leader were able to direct nearly wild-type expression. We have identified three separable control regions in the ninaE promoter. The first, which has the properties of an enhancer element, is located between nucleotides -501 and -219. The removal of this sequence had little effect on promoter function; this sequence appears to be redundant. However, it appears to be able to substitute for the second control region which is located between nucleotides -215 and -162, and which also affects the level of output from this promoter. Removal of these two control regions resulted in a 30-fold decrease in expression; however tissue specificity was not affected. The third control region, located downstream from nucleotide -120, appears to be absolutely necessary for promoter function in the absence of the first two regulatory sequences. Examination of larvae containing fusion genes expressing beta-galactosidase suggests that the ninaE gene is also expressed in a subset of cells in the larval photoreceptor organ. PMID- 3040525 TI - [Transfer of "artificial transposons" constructed on the basis of insertion element IS1]. AB - Terminal inverted repeats of the insertion element IS1 were synthesized chemically and plasmids containing these sequences flanking kanamycin-resistance gene in different combinations were constructed. Further incorporation of a whole sized copy of the IS1 into such plasmids caused in some cases the autonomous transfer of Km-resistance from plasmid to bacteriophage lambda DNA. The transposition of the Km-resistance gene was only observed in those cases when the gene was enclosed between IS1 copy and one of the terminal repeats. The data obtained are discussed with regard to the evolution of bacterial transposons. PMID- 3040526 TI - [Lysogenic conversion caused by phage phi 80. III. The mapping of the conversion gene and additional characterization of the phenomenon]. AB - The cor gene specifies lysogenic conversion caused by the lambdoid phage phi 80. The cor gene product inhibits tonA function in infected and lysogenic cells. The cells harboring pBR322 plasmid with the cloned cor gene of phi 80 became resistant to the phages T1 and phi 80 (TonA phenotype). The cor gene was mapped between 24 and 13 genes on the phi 80 phage genetic map. It is not essential for phage lytic growth. Its presence in lysogens leads up to accumulation of tonA mutants in a cell population. PMID- 3040527 TI - [Genetic similarity of various orthopedic anomalies]. AB - Genetic-mathematical analyses have proved that some orthopaedic anomalies are interconnected, have genetic communion, one of its components being the general dysplasia of connective tissue. PMID- 3040528 TI - [Creation of a clone panel of fox x Chinese hamster somatic cell hybrids and chromosome mapping of genes for LDHA, LDHB, GPI, ESD, G6PD, HPRT, alpha-GALA in the silver fox]. AB - A clone panel of fox-hamster somatic cell hybrids which can be used for fox gene mapping was set up. Analysis of patterns of chromosome-enzyme segregation made it possible to assign gene GPI to chromosome 1, LDHA to chromosome 11, LDHB to chromosome 8, ESD to chromosome 6 and G6PD, HPRT, alpha-GALA to chromosome X. PMID- 3040529 TI - Characterization of the Cephalosporium acremonium ribosomal RNA genes. AB - To investigate the organization of the ribosomal RNA genes in Cephalosporium acremonium, we cloned the whole r X DNA repeat in pBR322 and pNEO plasmids. Both the cloned and the genomic r X DNA fragments were characterized by restriction mapping. The r X DNA repeat unit was found to be 8.0 kb long and there was no significant heterogeneity among the individual repeats. PMID- 3040530 TI - Contingent replication assay (CRA) procedure for rapid isolation of enhancers. AB - A rapid procedure for the isolation of functional enhancer sequences consists of the construction of a shotgun DNA library in SV40-based plasmid shuttle vectors which depend on an enhancer for replication, the replication in monkey (CVI) cells of those vectors into which an enhancer sequence was inserted, the selective cleavage of unreplicated vectors by DpnI and the recovery of the replicated vectors by transfection into Escherichia coli. We describe conditions for the fusion of protoplasts to CVI cells, under which conditions the probability of only one type of plasmid entering a cell is increased and thus complementation and rescue of enhancer-less plasmids are decreased. The effectiveness of the procedure is demonstrated by the recovery of enhancers from bovine papillomavirus and Moloney murine sarcoma virus. PMID- 3040531 TI - Rapid identification of hybridization probes for chromosomal walking. AB - The presence of repeated elements in restriction fragments used as hybridization probes for chromosomal walking poses a major obstacle to the success of this gene cloning strategy. This report describes a simple and rapid means of identifying restriction fragments devoid of repeated sequences and therefore useful as hybridization probes for chromosomal walking. Restriction fragments derived from a genomic DNA clone are Southern blotted and hybridized to nick-translated total genomic [32P]DNA. Fragments of the genomic clone that contain high abundance sequences (i.e., repeated elements) hybridize strongly to their nick-translated counterparts, which, due to their high copy number, comprise a significant proportion of the total genomic DNA probe. Conversely, fragments containing single-copy or low-abundance sequences do not hybridize, as their nick-translated counterparts are poorly represented in the total genomic DNA probe. These latter fragments, by virtue of their low-abundance sequences, are well suited as probes for chromosomal walking. Ensuring the absence of repeated elements in restriction fragments prior to their purification and utilization as chromosomal walking probes results in marked savings of time, effort and materials. PMID- 3040532 TI - Expression of a major bovine rotavirus neutralisation antigen (VP7c) in Escherichia coli. AB - Sequences from genomic RNA segment 8 of the United Kingdom tissue-culture (t.c.) adapted bovine rotavirus encoding a major viral neutralisation antigen VP7c have been expressed in Escherichia coli. Expression under the regulated control of the bacteriophage lambda pR promoter was as a C-terminal extension to E. coli beta galactosidase (beta Gal). Following temperature induction, high levels of the fusion protein were synthesised and accumulated in induced cells, making up 5% 15% of total bacterial cell protein after 2 h of induction. Immunisation of sero negative rabbits and mice with gel-purified fusion-protein raised antibodies, which gave specific immunofluorescence with virus-infected cells and were able to immunoprecipitate proteins of the VP7 complex from such cells. Hyperimmune sera also gave a virus-type-specific reaction in a solid-phase enzyme-linked immunoabsorbant assay and neutralised virus infectivity in standard plaque reduction assays. PMID- 3040533 TI - [Fibrogenic activity of superhigh-siliceous zeolites]. PMID- 3040534 TI - [Concentration of various mineral elements in the placental tissue of women engaged in the production of mineral fertilizers]. PMID- 3040535 TI - Regulation of oxygen radical release from murine peritoneal macrophages by pharmacologic doses of PGE2. AB - The ability of pharmacologic doses of PGE2 to alter the release of superoxide (O2 ) and hydrogen peroxide (H2O2) from elicited peritoneal macrophages (M theta) was studied. Twice-daily administration of 200 or 100 micrograms of PGE2 to mice during accumulation of peritoneal M theta resulted in a significant reduction in M theta recovery and in the triggered release of H2O2, but not O2-. Cultivation of elicited M theta from normal mice with concentrations of PGE2 in excess of 10( 7) M for 24-48 h resulted in a significant reduction in the triggered release of H2O2, but not O2-. Cultivation for shorter periods of time or with lower concentrations of PGE2 failed to alter H2O2 release. This effect of PGE2 was reproduced by the phosphodiesterase inhibitor theophylline. The ability of PGE2 to inhibit H2O2 release in the presence of normal production of O2- was not prevented by the addition of superoxide dismutase. Cultivation of peritoneal M theta with 10(-5) M PGE2 for 48 h failed to increase intracellular catalase, although increased H2O2 scavenger activity was demonstrated. The inhibition of extracellular release of H2O2, but not O2-, by pharmacologic doses of PGE2 may be one mechanism for the anti-inflammatory action of this compound. PMID- 3040536 TI - Photolysis of nitrosamines and nitrosamides at neutral pH: a spin-trap study. AB - A model system has been used to study the types of radicals formed on denitrosation of N-nitroso compounds. Free radicals were formed at room temperature (22 degrees-23 degrees C) and neutral pH by photolytic cleavage of N nitroso bonds and were partially characterized following their addition to the spin traps 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) and N-tert-butyl-alpha-phenyl nitrone (PBN). Carbon-centered radical adducts were obtained during nitrosamine photolysis and nitrogen-centered radical adducts during nitrosamide photolysis. Since both the nitrosamines and nitrosamides initially form nitrogen-centered radicals on photolysis, a secondary reaction or rearrangement must occur after initial N-nitroso bond cleavage in the nitrosamines. Mechanisms are proposed to account for these results. PMID- 3040537 TI - The ability of scavengers to distinguish OH. production in the iron-catalyzed Haber-Weiss reaction: comparison of four assays for OH. AB - Kinetic analysis has been used to access how well scavenger inhibition can characterize the reactivity of oxidants produced in the iron-catalyzed reaction of H2O2 with xanthine oxidase-derived O2-.. Formate oxidation to CO2, deoxyribose oxidation, benzoate hydroxylation, and ethylene production from alpha-keto-gamma methiolbutyric acid (KMB) were measured. With Fe(EDTA) as catalyst, inhibition by most scavengers was quantitatively as expected for OH. involvement. Exceptions were urate and thiourea, which inhibited excessively and appeared to scavenge intermediates of the detection reactions. With nonchelated iron, there was minimal formate oxidation, but benzoate, KMB, and deoxyribose gave, respectively, 17%, 25%, and approximately the same product yield as with Fe(EDTA). Deoxyribose oxidation was not inhibited by some scavengers and excessively inhibited by others. However, scavengers that did not inhibit deoxyribose oxidation did inhibit with KMB and benzoate, and differences in scavenger effects in the presence and absence of EDTA in these assays were relatively minor. The results with formate and deoxyribose, but not KMB and benzoate, can therefore exclude free OH. as a significant oxidant product of the nonchelated iron-catalyzed Haber Weiss reaction. It is proposed that the different patterns of scavenger inhibition arise in the different assays because scavengers can react with intermediates in the detection reactions, all of which are multistep chains. Thus, inhibition may not signify OH. involvement, and similarities with inhibition expected for OH. my be fortuitous. PMID- 3040538 TI - Free radical modification of low-density lipoprotein: mechanisms and biological consequences. AB - Low-density lipoprotein readily undergoes lipid peroxidation that is accompanied by apoprotein fragmentation. Oxidized forms of low-density lipoprotein show altered biological behavior, including changes in receptor recognition and cytotoxicity to cells in culture. In this review, free radical mechanisms and the biological consequences of low-density lipoprotein modification are discussed. PMID- 3040539 TI - Uptake and distribution of cis-unsaturated fatty acids and their effect on free radical generation in normal and tumor cells in vitro. AB - We have previously shown that cis-unsaturated fatty acids (c-UFAs) possess a selective tumoricidal action that can be blocked by antioxidants. This property of c-UFAs might be due to various factors, including increased uptake, unusual distribution, or an ability to alter free radical generation in tumor but not normal cells. 14C-labelled linoleic acid (LA) uptake was almost the same in normal and tumor cells, whereas that of 14C-labelled arachidonic acid (AA) and 14C-labelled eicosapentaenoic acid (EPA) in tumor cells was substantially less than in normal cells. Tumor cells incorporate major portions of the fatty acids in the ether lipid and phospholipid fractions, whereas normal cells incorporate the fatty acids primarily in the phospholipid fraction. LA, AA, and EPA augmented nitroblue tetrazolium reduction, an indication of free radical generation, selectively in the tumor cells. These results suggest that there are significant differences between normal and tumor cells in fatty acid uptake and distribution, and in the ability of fatty acids to generate free radicals. PMID- 3040540 TI - Comparison of urea space, deuterium oxide space and body composition in growing pigs. AB - Urea and deuterium oxide (D2O) space were compared by simultaneous infusion into pigs weighing approximately 55 or 90 kg. Urea was cleared from the plasma pool more rapidly than D2O and appeared to equilibrate at a relative concentration which was lower than that of D2O. Consequently, urea and D2O space values were closest when extrapolated to zero time values. Correlations between urea space and D2O space were highest at 15 minutes post infusion (R2 = .75) or between urea space at 15 minutes and D2O at equilibrium (R2 = .86). Results of urea and D2O space measurements were compared with water, lipid and protein content of the carcass. Urea space at 15 minutes and D2O space at 35 minutes most closely approximated total body water while D2O space at 15 minutes was nearly equivalent to empty body water. Overall, D2O space at equilibrium had the highest correlations with carcass values of water, lipid and protein and appears to be preferable to urea space for estimating carcass composition in pigs. PMID- 3040541 TI - Small intestinal response to 'elemental' and 'complete' liquid feeds in the rat: effect of dietary bulk. AB - The effect of oral isocaloric feeding on small intestinal structure and function was studied in the rat. The liquid 'elemental' enteral feed Vivonex HN, the liquid 'complete' feed Ensure and the same liquid complete feed with 9% bulk Enrich were compared with solid chow containing 21% bulk (normal rat chow), all given for four weeks. Weight gain was significantly less in the group fed Vivonex HN than that of any other groups. The bulkless Vivonex HN and Ensure increased proximal jejunal mass compared to Enrich with 9% bulk or to normal rat chow. Jejunal mucosal DNA and protein levels also tended to be higher in Ensure and Vivonex HN fed animals, as was jejunal sugar absorption. In the terminal ileum, however, total weight was decreased by both elemental and complete feeds with and without bulk, but particularly by the elemental diet. Bulkless feeds therefore increase jejunal and reduce terminal ileal mass. The striking atrophy of the terminal ileum produced by the elemental diet may be important for its efficacy in treating inflammatory bowel disease. PMID- 3040542 TI - Vagal cholinergic control of gastric alkaline secretion in normal subjects and duodenal ulcer patients. AB - Gastric alkaline secretion was determined in ranitidine treated healthy subjects and duodenal ulcer (DU) patients using gastric perfusion aspiration system and back titration of gastric perfusate to original pH 6.0. Basal alkaline secretion showed periodic fluctuations reaching peaks at phase III of the migrating motor complex (MMC) in the stomach. Mean basal alkaline secretion in healthy normals and DU patients averaged 1120 +/- 124 and 880 +/- 72 mumol/h, respectively and no correlation was found between basal and maximally stimulated gastric acid and alkaline secretion. Modified sham feeding in normal subjects and in DU patients increased this secretion to the peaks of about 28 and 36% of the maximal alkaline response to intragastric application of 16,16 dimethyl-PGE2 in these subjects. Vagotomy did not affect significantly basal alkaline secretion but prevented the rise in alkaline secretion induced by modified sham feeding. Atropine (5-20 micrograms/kg) decreased dose dependently basal, and prevented the modified sham feeding induced alkaline secretion, while pirenzepine (5-20 micrograms/kg) had little influence on basal, and did not affect the modified sham feeding induced, alkaline secretion. This study shows that basal gastric alkaline secretion fluctuates in phase with gastric motor activity, and is similar in normal and DU patients. Vagal stimulation strongly increases alkaline secretion, the effect being abolished by vagotomy and atropine, but not by pirenzepine, suggesting the involvement of M2 rather than M1 subtypes of muscarinic receptors in this stimulation. PMID- 3040543 TI - Activation of peripheral blood and intestinal lamina propria lymphocytes in Crohn's disease. In vivo state of activation and in vitro response to stimulation as defined by the expression of early activation antigens. AB - In the present study the state of activation of either peripheral blood and intestinal lamina propria mononuclear cells in Crohn's disease was defined by investigating the expression of early activation antigens (namely the 4F2 antigen, the transferrin receptor and the interleukin-2 receptor). The expression of 4F2 and T9 antigens was greatly increased--in the peripheral blood and in the intestinal lamina propria whereas the proportion of interleukin-2 receptor bearing cells was much less pronounced. The counts of early activation antigens bearing cells in the lamina propria were quite comparable with those of the autologous peripheral cells. In the peripheral blood counts of 4F2 and T9 positive cells were very high in patients with active Crohn's disease but patients with quiescent disease also had a significantly raised proportion of 4F2 and T9 bearing cells. Only in those patients with no evidence of macroscopic disease (namely those resected without recurrence) the counts of early activation antigens bearing cells were within the normal range. The in vitro mitogen induced expression of early activation antigens on either peripheral and intestinal mononuclear cells of patients with Crohn's disease proved to be both quantitatively and qualitatively similar to that of the controls showing the full expression of 4F2, transferrin receptor, and interleukin-2 receptor. While demonstrating that in Crohn's disease there was no intrinsic defect of generation and expression of growth factors receptors by peripheral and intestinal lymphocytes, these results showed that there was a divergence in the expression of early activation antigens in vivo and in vitro. This would indicate that in Crohn's disease there is an in vivo increased population of preactivated rather than fully activated lymphocytes consisting of 4F2 and T9 bearing cells. The high proportion of these cells in the peripheral blood and in the intestine suggests that a chronic immune activation is present in these patients outside as well as within the affected bowel. PMID- 3040545 TI - Detection of human papillomavirus DNA in advanced epithelial ovarian carcinoma. AB - Tissue specimens from 10 out of 12 patients with advanced epithelial ovarian adenocarcinoma contained DNA of human papillomavirus type 6 (HPV-6). HPV DNA was identified by in situ hybridization at high stringency using biotin-labeled DNA probes. Nonneoplastic tissue specimens from other pelvic sites of the same patients were also examined. None showed evidence of HPV DNA. The meaning of these findings in relation to epithelial ovarian carcinoma is discussed. PMID- 3040546 TI - Induction of early myeloperoxidase in acute unclassified leukemia. PMID- 3040544 TI - Morphological and cell kinetic effects of dietary manipulation during colorectal carcinogenesis. AB - The effect of dietary manipulation of fat and fibre on the structural and cell kinetic characteristics of colonic mucosa was studied before and during experimental carcinogenesis in 232 male Albino Swiss rats. Carcinogen treated animals were given 12 weekly injections of azoxymethane (10 mg/kg/week). The animals were divided between four dietary groups (1) high fat, high fibre, (2) low fat, high fibre, (3) high fat, low fibre and (4) low fat, low fibre. Pathological and cell kinetic information together with details of certain faecal characteristics was collected when the animals were killed 4, 20, and 28 weeks after starting their experimental diet. Tumour induction was significantly influenced by diet. The highest risk of colorectal tumour development was found in groups fed diet 3: high fat, low fibre (p less than 0.03). In contrast, diet 2: low fat, high fibre was associated with the lowest risk. The proportion of histologically proven colonic tumours occurring in each dietary group was: diet 1 10.9%, diet 2-3.6%, diet 3-63.7%, diet 4-21.8%. Scanning electron microscopic (SEM) studies done on selected samples indicated both dietary and azoxymethane related alterations in crypt unit integrity. The most marked surface architectural changes were seen in carcinogen treated animals maintained on diet 3 (high fat, low fibre). Stathmokinetic analysis revealed considerable intergroup variability. Both fat and fibre produced significant effects, principally during the preneoplastic phase of carcinogenesis. Faster proliferative activity tended to be found in animals at low risk of tumour induction (diet 2), slower proliferation being more characteristic of animals at high risk (p less than 0.05). The findings suggest that both topographical and cell kinetic parameters have an important relationship with promoting and protecting dietary factors during the development of colorectal cancer. PMID- 3040547 TI - Cytomegalovirus hyperimmunoglobulin and substitution with blood products from antibody-negative donors. A pilot study in bone marrow transplant recipients. PMID- 3040548 TI - Prophylactic application of an anti-cytomegalovirus hyperimmunoglobulin in allogeneic bone marrow transplant recipients. PMID- 3040549 TI - Adrenergic receptors in inner and outer layers of human myometrium near term: characterization of beta-adrenergic receptor sites by [125I]-iodocyanopindolol binding. AB - The beta-adrenergic receptors present in inner and outer layers of human myometrium near term were characterized using the radioiodinated antagonist iodocyanopindolol (ICYP). In both layers ICYP binding is saturable, rapidly reversible, stereoselective, and appears to occur in a single class of sites with a KD of 30 pmol/l. Adrenergic agonists compete for ICYP binding sites with an order of potency consistent with beta 2-adrenergic potencies: isoproterenol greater than epinephrine much greater than norepinephrine. Studies in which CGP 20712 A, a beta 1-adrenergic antagonist, competes for ICYP binding sites in human myometrium reveal that at least 65% of the beta-receptors present are beta 2 subtype, whatever the layer considered. At the 35th week of pregnancy, the density of beta-adrenergic receptors in the inner layer (15.2 fmol/mg of protein) is about 50% higher than in the outer layer. At term, the densities of beta adrenergic receptors are reduced and exhibit the same values for both layers (5 fmol/mg of protein). These results indicate that the beta-adrenergic receptors in the two layers diminish during pregnancy and reach, at term, an equal and low level of density. PMID- 3040550 TI - Immunologic status, granulocyte chemotaxis and antibodies to HTLVIII/LAV in haemophiliacs treated with clotting factor concentrates. PMID- 3040551 TI - Molecular study of the Philadelphia translocation in chronic myelogenous leukemia in different stages of disease. PMID- 3040552 TI - [Juvenile postnasal angiofibroma]. PMID- 3040553 TI - [Carcinoma of the left breast, malignant melanoma]. PMID- 3040554 TI - [Effect of lead and cadmium ions on the ATPase activity of the human erythrocyte membrane]. PMID- 3040555 TI - [Studies of phosphorylation in rat mast cells (supplement 2). Diphosphoinositide kinase in rat mast cell granules]. AB - Rat mast cells were purified by a Percoll gradient, and the granules were isolated from the sonicated mast cells on a Percoll gradient. The granules were shown to contain a diphosphoinositide kinase which catalyzes the formation of triphosphoinositide (TPI) from diphosphoinositide (DPI). The enzyme requires ATP and Mg2+ for the activity. TPI formation is almost completely dependent on MgCl2 or MnCl2, and maximal response is observed at 20 mM or 1 mM, respectively. The Km for ATP is 3 microM. TPI formation in the granules is dependent on the reaction time. The maximal response is seen at 23 degrees C. NaCl, KCl, Na2HPO4 and KH2PO4 have no effect on the activity. One hundred microM adenosine, AMP, ADP, and 10 microM cyclic AMP inhibit the kinase activity. PMID- 3040556 TI - Tn1000 insertional mutagenesis of cloned repressor gene of the phage L: plasmid oligomerization in the presence of F'lac. AB - An ampicillin resistance plasmid carrying the cloned repressor gene cII of the L phage (Salmonella typhimurium) was conducted by F'lac into an F- recipient. Two types of plasmids were isolated from Apr transconjugants. The majority of plasmids were dimers with one copy of Tn1000 inserted, the minority being monomers with one copy of Tn1000. This proportion remained unaltered when we used the F'lac strain transformed with a monomeric form of the recombinant plasmid as a donor. An extensive oligomerization of pBR322-originating plasmids was proved in the presence of F'lac; its presumable relationship to transposition-related processes is suggested. PMID- 3040557 TI - [Duplex sonography of the carotid artery following neck dissection. Initial experiences in tumor after care]. PMID- 3040558 TI - [Objective evaluation of cataract development under treatment with cytochrome C, sodium succinate, adenosine, nicotinamide and sorbitol]. PMID- 3040559 TI - Emerging patterns in transitional care. PMID- 3040560 TI - Insulin-sensitive phosphodiesterase in fat cells from human subcutaneous adipose tissue. PMID- 3040561 TI - Effect of alpha-1 adrenoceptor blockade on plasma levels of atrial natriuretic peptide during dynamic exercise in normal man. PMID- 3040563 TI - Mucoid carcinomas of breast. Typing: exocrine, endocrine and amphicrine. PMID- 3040562 TI - Defective diurnal changes of food intake, plasma glucose and insulin in rats with a transplantable islet cell tumour. AB - Subcutaneous implantation of small fragments of a radiation-induced transplantable rat insulinoma into the subscapular region of 16- to 17-week-old male NEDH rats resulted, over a 22-day period, in the progressive development of marked hyperinsulinaemia and severe hypoglycaemia, despite a compensatory increase in food intake. Diurnal changes were examined at 3-hourly intervals for 24 h in control rats and tumour-bearing rats at 20-21 days after transplantation. The control animals exhibited distinct diurnal changes of food intake, glucose and insulin concentrations. Food intake was greatest between 17.00 and 23.00 h; plasma insulin was greatest between 20.00 and 23.00 h, and plasma glucose was raised at 20.00, 02.00 and 05.00 h, compared with the other times. In contrast, insulinoma-bearing rats displayed no diurnal changes other than a small decrease in food intake between 05.00 and 11.00 h. Plasma glucose and insulin concentrations were significantly different from control rats at all times, and food intake was significantly increased between 23.00 and 17.00 h. These observations demonstrate that the transplantable insulinoma not only causes hyperinsulinaemia and hypoglycaemia but results in hyperphagia and defective diurnal changes of food intake, plasma glucose and insulin concentrations. Interruption of nutrient intake by withdrawal of food for 6 h exacerbated the hypoglycaemia of insulinoma-bearing rats leading to coma. PMID- 3040564 TI - Human lung tumours: does intermediate filament co-expression correlate with other morphological or immunocytochemical features? AB - Co-expression of intermediate filaments is being increasingly reported for many human tumours including carcinoma of the lung. However, it is unclear whether such findings are unusual or restricted to a group of highly atypical tumours. In the present study the pattern of co-expression of intermediate filaments in 94 human lung tumours has been correlated with light and electron microscopical features which are thought to be atypical for particular tumour types. These same aberrant patterns of intermediate filament co-expression have also been compared with the proliferative rate of these tumours as determined by labelling with the monoclonal antibody Ki67. The results of this study have shown that the aberrant expression of intermediate filaments is not a feature unique to a group of highly unusual tumours but is found throughout the spectrum of lung cancer. The implications of these findings for the use of anti-intermediate filament antibodies in pulmonary pathology are discussed with suggestions for future directions which might be taken in this field. PMID- 3040566 TI - Correlation between immunohistochemically determined oestrogen receptor content, using monoclonal antibodies, and qualitative and quantitative tissue features in ductal breast cancer. AB - Previous studies have shown that oestrogen receptor content in breast cancer was correlated with qualitative and also, more strongly, with quantitative nuclear features in tissue sections. However, even with the better reproducible quantitative microscopical assessments, the variance in the correlation with oestrogen receptor was considerable. This might be due to the implicit problems of oestrogen receptor determination with the biochemical assay. Therefore, receptor content was studied using monoclonal antibodies in 50 consecutive invasive ductal breast cancers. Oestrogen receptor status was compared with qualitative features and with the mean and standard deviation of the nuclear area, morphometrically evaluated on immunostained and adjacent haematoxylin and eosin stained sections. In agreement with earlier observations, nearly all tumours with prominent elastosis were oestrogen receptor positive; but a minority of negative cases also showed elastosis. The correlation between the other qualitative features and receptor status was weak. A significant inverse correlation (P less than 0.001) existed between the receptor status and the mean and standard deviation of the nuclear area. Even with the highly reproducible morphometrical analysis, correlation between nuclear oestrogen receptor content and quantitative nuclear features was relatively weak. This might indicate that receptor status and nuclear morphometric features reflect different biological characteristics of breast cancers. PMID- 3040565 TI - Genotypic analysis of large cell lymphomas which express the Ki-1 antigen. AB - The monoclonal antibody Ki-1 reacts with Reed-Sternberg cells in Hodgkin's disease and with the tumour cells in a minority of large cell non-Hodgkin's lymphomas. This study describes the results of immunophenotypic and DNA analysis in 30 cases of non-Hodgkin's lymphoma, all of which expressed the Ki-1 antigen. The genotypic analysis has been undertaken using both immunoglobulin and T-cell receptor gene probes. Sixteen cases were shown by this method to be of monoclonal T-cell origin, six of B-cell origin, while in eight cases there was no evidence of either T- or B-cell lineage. This confirms previous immunohistological data indicating that non-Hodgkin's lymphomas which express the Ki-1 antigen may be of either T-cell or B-cell origin. PMID- 3040567 TI - Phyllodes tumour of the male breast. PMID- 3040568 TI - [The construction of a Chinese mitochondrial DNA library]. PMID- 3040569 TI - [A comparative study of sensitivity of the PAP, double-bridge PAP and ABC methods with four antibodies in a moist chamber and on a hotplate]. PMID- 3040570 TI - [A study on CMV shedding in 247 cases of breast milk]. PMID- 3040571 TI - [Chromium content of the hair of patients with acute cerebrovascular diseases]. PMID- 3040572 TI - [Primary malignant fibrous histiocytoma of bone. Clinico-pathologic analysis of 45 cases]. PMID- 3040573 TI - Identification of human papillomavirus types in male urethral condylomata acuminata by in situ hybridization. AB - An in situ hybridization technique was applied under stringent conditions to paraffin sections of urethral condylomata from male patients to determine the presence of DNA sequences of human papillomavirus (HPV) types 6, 11, 16, and 18. The material consisted of 15 classical condylomata acuminata, two flat condylomata, and five recurrent lesions. HPV DNA sequences could be identified in all 15 condylomata acuminata; in 13 lesions, two types of viral DNA were observed (types 6 and 11 in 12, types 6 and 18 in one). In the remaining two condylomata acuminata, only HPV type 11 was present. One of the two flat condylomata was negative with all the probes, and one was borderline-positive for HPV 6. Four of five recurrent lesions contained the same types of viral DNA as the primary lesions, albeit with slight differences in the intensity of viral expression. One lesion was negative with all probes. We conclude that urethral condylomata in males contain the same types of HPV as seen in other anogenital lesions of both sexes and that infection with two viral types is common. In situ hybridization with HPV DNA probes is applicable to archival material and therefore may prove to be of value in future epidemiologic studies comparing lesions in sexual partners. The determination of viral type may have therapeutic implications. PMID- 3040574 TI - Solid glomus tumor of the pterygoid fossa: a lesion mimicking an epithelial neoplasm of low-grade malignancy. AB - An example of a solid glomus tumor occurring in the pterygoid fossa is described. Despite its epithelial appearance on light microscopy, the tumor was of connective tissue origin. The diagnosis of glomus tumor was based on characteristic cytologic, ultrastructural, and immunohistochemical features, but the unusual site of the tumor made the diagnosis difficult. PMID- 3040575 TI - Cytoplasmic cytomegalovirus inclusions in human bile duct epithelia. PMID- 3040576 TI - Lack of association and linkage between HLA and familial polyposis coli. AB - We investigated possible association of and linkage between HLA and familial polyposis coli (FPC). In 182 individuals from 66 pedigrees of FPC and 108 individuals from a normal population, HLA-A, -B, and -C antigens were determined. When the frequencies of HLA antigens in 66 unrelated patients and in normal controls were compared, no association of FPC with HLA was observed. For the linkage analysis, HLA haplotypes of 17 affected sib pairs were investigated by the affected sib pair method. The number of pairs which shared two, one, and no haplotypes identical by descent was not significantly different from the number expected with random occurrence (P greater than 0.95). Finally, seven families were analyzed using Morton's sequential test. A maximum lod score of -0.056 at a recombination fraction of 0.4, and a lod of -3.089 at a recombination fraction of 0.05 were obtained. Therefore, there is neither an association of nor linkage between FPC and HLA. PMID- 3040577 TI - A small deletion in the Duchenne/Becker muscular dystrophy locus--a functionally important region? AB - A DNA deletion in a patient with Becker muscular dystrophy (BMD) has been delineated by restriction endonuclease mapping. The deletion is unusually small, removing six kilobases (kb) of DNA distal to pERT 87-1 (DXS164). This region has previously been shown to contain an exon of a candidate gene which, when defective, causes Duchenne muscular dystrophy (DMD) or Becker muscular dystrophy. Removal of this exon and surrounding DNA is apparently sufficient, in this case, to cause a BMD phenotype. The occurrence of this deletion in DXS164 would appear to confirm that this region is part of the BMD locus. Many DMD patients have deletions in and around this region, adding further evidence for the allelic nature of the two disorders. This fortuitous deletion may identify a functionally important domain of the protein product in terms of the severity of phenotype manifested. PMID- 3040578 TI - Self-poisoning and therapeutic intoxication with lithium. AB - Of 68 admissions for lithium overdose over 16 years, 25 were due to therapeutic intoxication and 43 to deliberate self-poisoning. Three patients with therapeutic intoxication had acute diabetes insipidus with hypernatraemia. One of them had acute renal failure requiring dialysis, prolonged Parkinsonism and generalised myopathy. Twenty-two patients with therapeutic intoxication had peak serum lithium concentrations above the therapeutic range. In contrast, of 22 self poisoned patients with peak serum lithium concentrations above the therapeutic range only 3 developed toxicity. The mean admission plasma urea concentration in patients with therapeutic intoxication was higher than in self-poisoned patients and the mean admission plasma bicarbonate concentration was lower. The mean serum lithium half-life in 8 patients with therapeutic intoxication was considerably longer than in 5 self-poisoned patients. Renal lithium clearance is enhanced by increased sodium excretion and we recommend that lithium toxicity be treated with saline diuresis and frusemide if fluid retention occurs. Haemodialysis is mandatory when renal failure is present, and may be indicated when serum lithium concentrations are very high or rising rapidly. PMID- 3040579 TI - A novel approach for the production of monoclonal antibodies using infectious vaccinia virus recombinants. AB - We describe a novel approach of producing monoclonal antibodies (MABs) to one specific protein of a virus or other agent consisting of several proteins, without the use of purified antigen in either the immunization or screening phase of the procedure. This method has general application in the production of MABs when the antigen cannot be obtained in a pure form, but the gene is available. We illustrate this application by producing MAB specific to the nucleocapsid protein (N) of vesicular stomatitis virus serotype Indiana (VSV-IN) from BALB/c mice immunized with an infectious vaccinia virus recombinant vector (v38) that expresses the N gene of VSV-IN. This novel method of immunization obviates the need for initial purification of the protein antigen and injection of adjuvants with the isolated protein as is done in traditional MAB production. PMID- 3040581 TI - Infrared study on the protonation of Schiff base with tyrosine: a model study for bacteriorhodopsin chromophore. PMID- 3040580 TI - Production of monoclonal antibodies specific to theophylline-treated lymphocytes. AB - The T11 molecule is reported to play a key role in T lymphocytes activation. Moreover, theophylline is known to modify the functional properties of T lymphocytes probably inducing early changes in T11 molecule during T cell activation. Aim of our work was to clarify the effect on T lymphocyte surface after in vitro treatment with theophylline. In this paper, we describe the production and the functional properties of several monoclonal antibodies obtained by immunizing Balb/c mice with theophylline treated cells. Some of the monoclonal antibodies reacted only with the theophylline-treated lymphocytes and showed a promitogenic activity, enhancing the expression of T activated cell antigen. These monoclonal antibodies seem to demonstrate the existence of a membrane molecule which appears on lymphocytes surface after a trigger signal occurring in the early stages of T cell activation likely related to the T11 dependent alternative pathway. PMID- 3040582 TI - [Endocrine rhythms in patients with rheumatoid arthritis: possible neurohormonal influence on immunological reactions]. AB - Recent studies are reporting on an influence of ACTH and prolactin in physiological dosages on the intensity of the immune response in the animal model of adjuvant arthritis. Therefore, we studied the circadian secretion patterns of ACTH, cortisol and prolactin by measurements throughout a 24-hour cycle in two hour intervals in patients with rheumatoid arthritis with different inflammatory activity of the disease. We only investigated patients who never before were treated with corticosteroids and drugs like gold, d-penicillamine or immunosuppressive drugs. The circadian secretion patterns of cortisol and ACTH were disturbed in most patients. High inflammatory activity was accompanied by severe disturbance of the circadian rhythm, whereas patients with low inflammatory activity showed a nearly normal secretion pattern. Also the measurements of prolactin showed a tendency of loss of circadian rhythm in relation to the inflammatory activity of the disease. According to the found changes of endocrine regulation of ACTH, cortisol and prolactin, an influence of mediators of inflammation on hypothalamic centers, analogous to endogenous pyrogen in fever should be discussed. PMID- 3040583 TI - Stable expression of a selectable myeloproliferative sarcoma virus in murine T lymphocyte and monocyte cell lines. AB - We have investigated whether a retroviral vector based on the myeloproliferative sarcoma virus (MPSV) can be expressed in murine T cells and macrophages. This vector (neoR MPSV) carries the dominant selection marker for neomycin resistance (neoR) and the mos oncogene. The murine T cell line BW5147 and the monocytic cell line P388D1 were either transfected with neoR MPSV DNA or infected with neoR MPSV virus. From both lines, neoR cell clones could be established by retroviral infection, but not by calcium-phosphate precipitation-mediated DNA transfection. The efficiency of infection could be increased 60- to 200-fold upon cocultivation of target cells with irradiated neoR MPSV virus-producing cells. All neoR clones showed neoR MPSV specific sequences as revealed by dot blot and Southern blot analysis. The integration and expression of neoR MPSV was stable over a period of now more than 4 months, even in the absence of selection for neomycin resistance. Northern blot analysis showed that neoR clones express full length neoR MPSV. Further, clones of both T cell and monocyte origin were capable to produce infectious virus particles as revealed by focus formation on fibroblasts and conversion of neomycin sensitive fibroblasts to a neomycin resistant phenotype. PMID- 3040584 TI - Polyclonal activation of human peripheral blood lymphocytes by bacterial porins and defined porin fragments. AB - Bacterial porins were isolated from Escherichia coli B and Salmonella typhimurium S 1135. The proteins were cleaved either by cyanogen bromide treatment or by enzymatic digestion into a variety of small fragments, and the compounds were characterized by SDS-polyacrylamide gel electrophoresis. Both the porins and the porin fragments constituted potent mitogens for human peripheral blood lymphocytes, comparable to the human B-lymphocyte activator pokeweed mitogen. In the cultures, B-lymphocytes were stimulated into immunoglobulin production, as measured by ELISA. In all experiments, the activity of the mitogens extracted from S. typhimurium was superior to that of the compound isolated from E. coli B. The well-defined porins constitute valuable tools for investigating the molecular mechanism of human lymphocyte activation. PMID- 3040585 TI - Differences in H-2 recombinant mice in the beta-naphthoflavone inducibility of the mixed function monooxygenase, P1-450. AB - The influence of the major histocompatibility complex (H-2 in mouse) on induction of cytochrome P-450-dependent monooxygenase (P1-450) by the prototype polyaromatic hydrocarbon (PAH), beta-naphthoflavone, was investigated in C57BL/10 Sn (B10) recombinant congenic mice. The cytosolic Ah-receptor level, as measured by specific binding with [3H]-2,3,7,8-tetrachlorodibenzo-p-dioxin, was significantly lower in B10.A and B10.A(5R) than in either B10, B10.BR, or B10.A(2R), suggesting that the D region of H-2 influences Ah-receptor levels. The responsiveness to beta-naphthoflavone, as determined by increased catalytic activity toward benzo(a)pyrene and 7-ethoxycoumarin, was considerably lower in B10, B10.A, and B10.A(5R) than in B10.BR and somewhat lower than in B10.A(2R) or B10.A(4R) mice. The lower PAH responsiveness in B10.A and B10.A(5R) correlated with their lower Ah-receptor levels while that in B10 appeared to reflect a K-A region influence on PAH responsiveness that was not due to changed Ah-receptor levels. Thus, we conclude that more than one H-2 locus may influence PAH responsiveness, and by different mechanisms. PMID- 3040586 TI - Erythrocyte ghost Na+,K+-ATPase and blood pressure. AB - To examine the relationship between body mass index, blood pressure, and the Na+,K+-adenosine triphosphatase (ATPase) system, we measured the erythrocyte ghost Na+,K+-ATPase and the erythrocyte Na+ concentration in 120 blacks and 127 whites (136 males and 111 females). Blacks showed a 13.9% higher erythrocyte Na+ (7.63 +/- 0.19 vs 6.70 +/- 0.11 [SEM] mEq/L; p = 0.0001) and a 16.1% lower erythrocyte ghost Na+,K+-ATPase activity (140.3 +/- 4.2 vs 167.3 +/- 4.7 nmol inorganic phosphate/mg protein/hr; p = 0.0002) than whites. Male subjects demonstrated a 6.4% higher erythrocyte Na+ (7.35 +/- 0.17 vs 6.91 +/- 0.14 mEq/L; p = 0.043) and an 11.5% lower Na+,K+-ATPase activity (145.7 +/- 3.7 vs 164.7 +/- 5.5 nmol inorganic phosphate/mg protein/hr; p = 0.0015) than female subjects. Significant (p less than 0.001) negative correlations were identified for the systolic, diastolic, and mean blood pressure levels and the erythrocyte ghost Na+,K+-ATPase. These findings were complemented by positive correlations for the blood pressure levels and erythrocyte Na+ concentrations. The body mass index was negatively correlated with erythrocyte ghost Na+,K+-ATPase and it accounted for 6.7%, 5.6%, and 6.1% of the variabilities in the systolic, diastolic, and mean blood pressure levels, respectively. Variabilities of 1.4% systolic, 12.3% diastolic, and 11.1% in mean arterial pressure were attributable to the erythrocyte ghost Na+,K+-ATPase activity. Provided that findings in erythrocytes also reflect the relative status of the vascular smooth muscle cell Na+,K+ ATPase, the predisposition of black, male, and obese persons to hypertension may relate, among other factors, to a lower activity of this enzyme system, which results in an increased vascular tone. PMID- 3040587 TI - Evaluation with an iuc::Tn10 mutant of the role of aerobactin production in the virulence of Shigella flexneri. AB - To evaluate the role of aerobactin production in the virulence of Shigella flexneri, a iuc::Tn10 insertion mutant was obtained from strain M90T, a serotype 5 isolate. This mutant was tested for its ability to invade and kill HeLa cells in monolayers, to elicit keratoconjunctivitis in guinea pigs and to infect ligated segments of rabbit ileal loops. Although this mutant did not grow in iron depleted media, its ability to grow intracellulalry and eventually kill HeLa cells was unchanged from that of the wild-type strain. On the other hand, an inoculum-dependent effect was observed in the Sereny test, as well as in the rabbit ligated ileal loop model, which was monitored for fluid production and for both gross and microscopical alterations of the mucosa. Transduction of the mutation within a noninvasive plasmidless derivative of the parental strain did not alter growth within the intestinal lumen. We conclude that aerobactin production most probably provides invasive strains with a selective advantage for growth within tissues when located in extracellular compartments. PMID- 3040589 TI - Oxidative killing of Aspergillus fumigatus proceeds by parallel myeloperoxidase dependent and -independent pathways. AB - The relative importance of several oxygen intermediates in fungicidal action against opsonized Aspergillus fumigatus conidia was investigated with monocytes from normal volunteers and patients with either chronic granulomatous disease or myeloperoxidase (MPO) deficiency. Results from experiments in which catalase, taurine, mannitol, or glucose-glucose oxidase were added to these phagocytes indicated that the MPO-hydrogen peroxide-halide system and an MPO-independent oxidative system exerted comparable conidiacidal activity. These findings offer a plausible explanation for the susceptibility of patients with chronic granulomatous disease to invasive Aspergillus infections; their phagocytes fail to generate hydrogen peroxide, a substrate necessary for both systems. Patients with MPO deficiency are not known to be predisposed to invasive aspergillosis, suggesting that an MPO-independent oxidative system may provide an alternative mechanism for the oxidative killing of Aspergillus spp. PMID- 3040588 TI - Nucleotide sequence of the leukotoxin genes of Pasteurella haemolytica A1. AB - A 4.4-kilobase-pair DNA fragment coding for the leukotoxin of Pasteurella haemolytica A1 has been isolated, and its nucleotide sequence has been determined. Two open reading frames, designated lktC and lktA, coding for proteins of 19.8 and 101.9 kilodaltons, respectively, were identified. Expression of the two genes in minicell-labeling experiments resulted in the production of the predicted proteins LKTC and LKTA. By using an antiserum against the soluble antigens of P. haemolytica A1 in Western blot (immunoblot) analysis of total cellular proteins from the Escherichia coli clones, LKTA was identified as an additional antigenic protein. Results from subcloning of the DNA fragment suggested that expression from both lktC and lktA is required for leukotoxin activity, indicating that the leukotoxin of P. haemolytica A1 is encoded by two genes. A comparison of the organization and the DNA sequence of the leukotoxin genes with those of the E. coli alpha-hemolysin genes showed a significant degree of homology between the two loci. This analysis suggested that the leukotoxin genes of P. haemolytica A1 and the E. coli alpha-hemolysin genes may have evolved from a common ancestor and that the two toxins may share similar activities or functional domains or both. PMID- 3040590 TI - Bacterial antigens induce collagenase and prostaglandin E2 synthesis in human gingival fibroblasts through a primary effect on circulating mononuclear cells. AB - Our previous work suggests that one mechanism through which connective tissue breakdown might occur in periodontal diseases is the production of metalloproteinases, including collagenase, by gingival fibroblasts. In this study we investigated whether highly purified preparations of lipopolysaccharide (LPS) and lipoteichoic acid (LTA) from a number of putative periodontal pathogens could induce monolayer cultures of human gingival fibroblasts to synthesize collagenase and prostaglandin E2. Using both biochemical assays and immunocytochemical techniques, we found that cells synthesized only very small amounts of collagenase in direct response to LPS or LTA (0.1 to 20.0 micrograms/ml). At the highest dose of both antigens, prostaglandin E2 production was increased. We then studied whether LPS and LTA could signal collagenase production by interacting primarily with a different cell type. Our results show that LPS and LTA were each able to stimulate cultures of human blood mononuclear cells (greater than 95% monocytes) to release collagenase-inducing cytokines. By indirect immunocytochemistry, we found that a large proportion of human gingival fibroblasts was activated to produce collagenase by supernatants from LPS- and LTA-stimulated mononuclear cells, whereas gingival fibroblasts cultured with supernatants from unstimulated mononuclear cells were not. Furthermore, in a population of activated fibroblasts we demonstrated, using a double-labeling technique, that some cells made collagenase and the specific tissue inhibitor of metalloproteinases (TIMP) simultaneously. As yet, the collagenase-inducing signals remain poorly characterized but the interleukins-1 and tumor necrosis factors seem likely candidates. PMID- 3040592 TI - Localization of stx, a determinant essential for high-level production of shiga toxin by Shigella dysenteriae serotype 1, near pyrF and generation of stx transposon mutants. AB - Hfr strains of Shigella dysenteriae serotype 1 were constructed by transient integration of an RP4 plasmid derivative carrying transposon Tn501 into the Shigella chromosome through Tn501-mediated cointegration. The Hfr strains were mated with Escherichia coli K-12 recipients carrying various auxotrophic markers, and E. coli recombinants which had received prototrophic Shigella genes were selected. Some of the E. coli transconjugants produced high levels of a cytotoxin which was neutralized by both polyclonal and monoclonal anti-Shiga toxin sera. The determinant for Shiga toxin production, designated stx, was first transferred to E. coli K-12 and then mapped by Hfr crosses to the trp-pyrF region located at 30 min on the E. coli chromosome. Bacteriophage P1-mediated transduction analysis of stx gave the following gene order: trp-pyrF-stx. The level of Shiga toxin production in E. coli Stx+ transconjugants and transductants was as high as that of the parental S. dysenteriae 1 strain. Stx- mutants of an Stx+ E. coli transductant were generated by random in vivo insertion mutagenesis with a Tn10 derivative transposon, Tn-mini-kan, followed by P1 cotransduction of the kanamycin resistance and PyrF+ markers into a pyrF Stx+ E. coli K-12 recipient. One stx::Tn-mini-kan transposon mutation was transferred by P1 transduction from this E. coli Stx- mutant to an E. coli K-12 Hfr strain and in turn transferred by conjugation to the original S. dysenteriae 1 strain plus two others. All kanamycin-resistant recombinants of S. dysenteriae 1 had lost their ability to produce high levels of Shiga toxin. A gene that specifies high-level Shiga toxin production is thus located near pyrF on the chromosome of S. dysenteriae 1. Stx- mutants of S. dysenteriae 1 exhibited full virulence in the Sereny test. PMID- 3040591 TI - Molecular cloning and characterization of the glucosyltransferase C gene (gtfC) from Streptococcus mutans LM7. AB - A glucosyltransferase (GTF) gene, designated gtfC, was cloned from Streptococcus mutans LM7. Its gene product was detected by screening a bacteriophage lambda library with rabbit antiserum raised against S. mutans LM7 extracellular proteins. DNA isolated from the immunopositive recombinant phage revealed two S. mutans chromosomal EcoRI fragment inserts, 8.1 and 4.7 kilobase pairs in size. Escherichia coli minicell analyses revealed the approximate position and direction of transcription of the gtfC gene. The gene product was determined to be a polypeptide of about 150 kilodaltons which synthesized a water-soluble glucan. Restriction endonuclease mapping and DNA hybridization indicated a repeated region of DNA corresponding to a portion of the coding region of gtfC immediately downstream from the intact gtfC locus on the chromosome. A 300-base pair gtfC-specific probe showed that the gene and the putative duplicated sequence were present in S. mutans serotypes c, e, and f, but not in other related oral streptococci which had GTF activity. In addition, the gtfC determinant displayed homology to sequences corresponding to the carboxy-terminal coding region of a gene (gtfB) encoding a GTF activity that synthesized water insoluble glucans. These data suggest that at least one class of GTF genes may be present in multiple copies in S. mutans and, further, that GTF genes may contain conserved sequences internal to their coding regions. PMID- 3040593 TI - Outer membrane permeability of Acinetobacter calcoaceticus mediates susceptibility to rat polymorphonuclear leukocyte granule contents. AB - Growth of Acinetobacter calcoaceticus on specific alkanes altered the outer membrane permeability of the organism, as indicated by a change in sensitivity to the antibiotic actinomycin D. As the carbon length of the alkane energy source decreased, outer membrane permeability and susceptibility to actinomycin D increased. Concomitant with the increase in outer membrane permeability, A. calcoaceticus became more susceptible to the oxygen-independent antimicrobial activity of extracted contents from rat polymorphonuclear leukocyte granules. Individual fractions of granule extract possessed no antimicrobial activity against A. calcoaceticus. The alkane-induced change in outer membrane permeability was not associated with alterations of lipopolysaccharide O antigen. An outer membrane permeability mechanism, independent of changes in lipopolysaccharide content, mediating susceptibility to the oxygen-independent antimicrobial activity of rat polymorphonuclear leukocyte granule contents is suggested. PMID- 3040595 TI - Transposon mutagenesis of group B streptococcus beta-hemolysin biosynthesis. AB - Beta-hemolysin production by group B streptococci (GBS) is speculated to be a major virulence factor of the organism. A virulent, beta-hemolytic group B streptococcus strain was mutagenized with the self-conjugative transposon Tn916 to derive isogenic strains with mutations only in the gene(s) responsible for beta-hemolysin biosynthesis. There was no significant difference between the virulence of the parent strain and that of the mutant strains in a neonatal rat sepsis model. PMID- 3040596 TI - Toluene diisocyanate respiratory reactions. I. Reassessment of the problem. AB - A series of workers with exposure to and respiratory symptoms from toluene diisocyanate (TDI) was compared with a group of exposed, asymptomatic workers and with normal controls using immunologic and clinical evaluations. Improved understanding of TDI-induced respiratory disease may best be achieved by categorizing the symptomatic workers into at least 3 and possibly 4 groups: immunologic TDI asthma; pre-existing chronic respiratory disease exacerbated or unaffected by TDI; asthma coincidental with TDI exposure, and possibly TDI respiratory disease secondary to toxic or inflammatory mechanisms. PMID- 3040597 TI - Prognostic implications of expression of the cellular genes myc, fos, Ha-ras and Ki-ras in colon carcinoma. AB - Messenger RNA levels of the c-fos, c-myc, c-Ha-ras and c-Ki-ras genes were studied in 39 tissue samples obtained from 17 patients undergoing surgery for colon carcinoma and other colon diseases. DNA extracted from the same samples was studied by Southern analysis. The tissues were tumors and grossly normal mucosa from each case and in some instances benign polyps and metastases. Our results indicate: (1) that 50% of cases studied show an increase in expression of at least one of the oncogenes studied; (2) that over-expression is not random, some cases over-expressing several of the genes studied; (3) that the expression pattern of the oncogenes studied varies between primary tumor and metastases; (4) that amplification is a rare event, being limited to one instance in which c-myc was amplified in a metastasis; (5) that cases which exhibit high levels of mRNA in one or more genes studied correlate with biologically aggressive tumors; and (6) that "non-expressors" are at higher risk for local recurrence based on correlations with mucin histochemistry. PMID- 3040594 TI - Evidence for activation of a respiratory burst in the interaction of human neutrophils with Mycobacterium tuberculosis. AB - We examined the capacity of human neutrophils to develop a respiratory burst, as monitored by superoxide release, in response to interaction with Mycobacterium tuberculosis. Serum-opsonized, heat-killed mycobacteria induced significant release of superoxide from neutrophils after 30 min of exposure, with a maximum release of 34 +/- 1.7 nmol/30 min per 5 X 10(6) neutrophils occurring with a mycobacterium/neutrophil ratio of 40:1. Similar levels of superoxide release were induced by live mycobacteria. Neutrophil superoxide production was reduced significantly with exposure to unopsonized organisms or by substitution of heat inactivated serum for opsonization. Mycobacterial components including culture filtrate, purified protein derivative, and the cell wall polysaccharide arabinogalactan failed to induce significant release of superoxide from neutrophils. Transmission electron microscopy demonstrated that more than 90% of the neutrophils had ingested heat-killed mycobacteria concomitant with the development of respiratory burst activity. These data suggest that the presumed failure of neutrophil killing of mycobacteria cannot be attributed to a lack of phagocytosis or respiratory burst activation. PMID- 3040598 TI - Subcellular concentrations of estrone, estradiol, androstenedione and 17 beta hydroxysteroid dehydrogenase (17-beta-OH-SDH) activity in malignant and non malignant human breast tissues. AB - Total and subcellular (cytosol and nuclear) concentrations of estrone (E1), estradiol (E2), and androstenedione were determined in non-malignant (n = 61) and malignant (n = 65) human breast tissues obtained from post-menopausal women. The 17 beta-hydroxysteroid dehydrogenase (17 beta-OH-SDH) activity was determined in 800g supernatant fraction. Total estrogens, E1 and E2 levels and 17 beta-OH-SDH activity were significantly (p less than 0.005, 0.0005, 0.001, respectively) higher in malignant than in non-malignant breast tissues. We failed to observe significant changes in subcellular steroid concentrations or enzyme activity associated with patients' obesity or tumor estrogen receptor status. When the steroid levels were analyzed in relation to clinical staging of the disease, nuclear contents of estradiol were significantly higher (p less than 0.005) in Stage-IV patients than in those with less advanced disease (Stages I to III). 17 beta-OH-SDH activity was significantly (p less than 0.001) lower in patients with advanced disease than in those with relatively less advanced (Stages I to III) disease and was positively correlated with tissue concentration of androstenedione. Our present data indicate that differential intracellular metabolism of steroid hormones may have some influence on availability of estradiol at nuclear sites. In postmenopausal women, local interconversion of estrogens may provide sufficient estrogenic stimulus to enhance the growth and progression of breast tumors. PMID- 3040599 TI - Antibody response against the Epstein-Barr virus-coded nuclear antigen2 (EBNA2) in different groups of individuals. AB - Specific antibody responses against the 2 major subcomponents of EBNA, EBNA1 and EBNA2 were evaluated, in order to study whether this serological study was beneficial compared to classical EBV serology. During this investigation, 491 sera, obtained from blood donors and patients with Burkitt's lymphoma (BL), nasopharyngeal carcinoma (NPC), infectious mononucleosis (IM), Hodgkin's disease, renal transplantation, rheumatoid arthritis and Human Immunodeficiency Virus (HIV) infection, were tested. While the anti-EBNA1 response followed the classical anti-EBNA/Raji response (99% of anti-EBNA/Raji-positive sera also recognize EBNA1), the anti-EBNA2 response was much less frequent and did not correlate with either anti-EBNA/Raji or anti-EA antibodies. In a control population, 8% of individuals had antiEBNA2 antibodies at titers greater than or equal to 10. The percentage was 45% in NPC and 38% in EBV-associated BL; thus, although not detected in all patients with EBV-associated tumors, anti-EBNA2 serology might be a useful marker in BL and NPC. No antibody was detected in the early course of IM, but in rheumatoid arthritis and in HIV-infected patients, the percentage of positive individuals reached 54 and 68, respectively. Seroconversion to EBNA2 was noted in a few cases, including renal transplant recipients, AIDS patients, and complicated IM. This suggests that in these situations, EBNA 2 serology might represent a useful marker related to modulation of the immune status or EBV reactivation. PMID- 3040600 TI - Different behaviour of normal and neoplastic cells cultured in vitro in the presence of catalase and superoxide dismutase. AB - Chicken embryo fibroblasts and hepatocytes were studied in the presence of catalase and superoxide dismutase in order to establish whether these enzymes had the capacity to favour cell multiplication as previously shown for in vitro tumour ascites cells (ATP C+). The results indicate that, unlike ATP C+ cells, both fibroblasts and hepatocytes are inhibited in their multiplication by superoxide dismutase. Similar effects are exerted on hepatocytes by catalase, whereas the multiplication of fibroblasts is favoured by high doses of this enzyme. Enzyme determinations revealed high levels of catalase and superoxide dismutase in hepatocytes, whereas both enzymes were poor in fibroblasts and ATP C+. PMID- 3040601 TI - Down-regulation of the EBV-encoded membrane protein (LMP) in Burkitt lymphomas. AB - A radioimmunoassay (RIA) has been developed and used to determine the expression of LMP-a membrane protein encoded by the LT3 region of the Epstein-Barr virus (EBV) genome-in cell lines of various origins. The RIA was highly sensitive, specific and reproducible. All EBV-negative cell lines were LMP-negative and 18 of 21 EBV-carrying cells were LMP-positive. LMP concentrations varied widely, ranging approximately from less than 4 ng up to 650 ng/mg protein. In several instances comparisons were made between lymphoblastoid (LCLs) and Burkitt lymphoma (BL) cell lines (EBV-positive or EBV-converted sublines of originally EBV-negative BL) originating from the same patient. In all such cases LMP and LMP specific mRNA levels were higher in the LCLs. Most of the LMP was found in the cytosol fraction, yet this fraction was negative in immunoblotting tests. However, antiserum preincubated with the cytosol lost its ability to react in immunoblotting with membrane LMP, indicating that the 2 LMP forms (membrane and cytosol) are completely cross-reactive. PMID- 3040602 TI - MHC-controlled susceptibility to C3H-MTV-induced mouse mammary tumors is predominantly systemic rather than local. AB - The major histocompatibility complex (MHC) is one of the genetic factors involved in the control of susceptibility to MTV-induced mammary tumors in mice. Previously, genetic differences in mammary tumor susceptibility were shown to be mainly due to differences in susceptibility of the mammary gland itself. Such experiments, however, were carried out using inbred strains which differ in many genes. Therefore, the conclusion drawn from such experiments is not necessarily applicable to every gene involved. We tested whether in the strain C57BL/10 (B10, resistant) and the H-2 congeneic strain B10.A(5R) (5R, susceptible), which differ only at the MHC, the MHC-controlled difference in tumor susceptibility indeed resides in the mammary gland tissue itself, or is caused by systemic factors. Mammary glands of infant B10 and 5R mice were transplanted into mammary fat pads of C3H-MTV-infected mammectomized (B10 X 5R)F1 females. The hosts received a hypophyseal isograft under the kidney capsule or were subjected to force-breeding to provide hormonal stimulation necessary for mammary tumor development. Control groups of C3H-MTV-infected B10, 5R and (B10 X 5R)F1 females and (B10 X 5R)F1 females without C3H-MTV were also subjected to the 2 types of hormonal stimulation. There was no difference in susceptibility between the 5R and B10 mammary glands transplanted into C3H-MTV-infected F1 hybrid hosts. On the other hand, C3H-MTV-infected 5R and (B10 X 5R)F1 females were significantly more susceptible than the C3H-MTV-infected B10 females, when either of the 2 methods of hormonal stimulation was used. This indicates that, in the strains used, MHC controlled susceptibility to C3H-MTV-induced mammary tumorigenesis is predominantly or exclusively controlled by systemic factors rather than by factors residing in the target tissue. PMID- 3040604 TI - Activation of endogenous MMTV proviruses in murine mammary cancer induced by chemical carcinogen. AB - A study was undertaken to determine whether activation of expression of silent endogenous mouse mammary tumor virus (MMTV) proviruses may occur during tumor induction by a chemical carcinogen. A series of transplantable mammary tumors induced in BALB/c mice by treatment with dimethylbenz(alpha)anthracene (DMBA), pituitary isograft, or both was examined. The results obtained suggest that chemical carcinogens may induce mammary tumors through more than one pathway. Two of 9 tumor lines produced virus-specific products at levels above those observed during the course of normal mammary gland development. One tumor contained high levels of MMTV-specific envelope [3.8 kilobase (kb)] and genomic length (8.9 kb) RNAs. This tumor expressed core- and envelope-related proteins detectable by immunoblotting (including p28, gp52, and gp36), displayed an acquired provirus with a restriction map different from those of described exogenous MMTV strains, and contained abundant virus particles. The other tumor that expressed high levels of MMTV gene products contained envelope-specific (3.8 kb) and long terminal-repeat-specific (1.6 kb) messages but no full-length RNA. It exhibited an aberrant 39 kDa, envelope-related protein, but no virus particles. Methylation data implicated the usually silent endogenous Mtv-8 provirus as the source of the abnormal envelope protein. None of the tumors expressed RNA from the putative mammary oncogenes, int-1 or int-2. We propose that chemical carcinogens may activate different cellular genes by mutation and that, in a subset of DMBA induced mammary tumors, the target genes include endogenous MMTV proviruses that are normally not expressed. The effect on provirus expression varies from tumor to tumor, but is stable over passage of a given tumor. MMTV may be of etiological importance in the genesis of those DMBA-induced tumors which contain high levels of MMTV-specific products, but its action in the BALB/c system is not mediated through enhanced expression of the int-1 or int-2 preferred integration regions. PMID- 3040603 TI - Stable transfection of a human lymphoma line by sub-genomic fragments of Epstein Barr virus DNA to measure humoral and cellular immunity to the corresponding proteins. AB - An Epstein-Barr virus (EBV)-negative human lymphoid B-cell line, DG75, was stably transfected with recombinant selection vectors that carry a subfragment of the BamHI WYH region (nucleotides 44664 to 50628), the BamHI K fragment, or a subfragment of the EcoRI D region (nucleotides 166614 to 170149) of B95-8 EBV DNA. These fragments contain the coding exons for the EBV-determined nuclear antigens EBNA2 and EBNA1, and the membrane antigen LMP, respectively. Antigen expression of the cells was detected by immunofluorescence. EBNA2 was expressed in 80-100% of the transfected cells, in contrast to EBNA1 which was expressed in only 25%, and LMP in only about 5% of the cells. Humoral antibody responses were measured by immunofluorescence and compared to cellular immunity as determined by the leukocyte migration inhibition (LMI) technique. Extracts from transfected cell lines expressing EBNA1, EBNA2 or LMP elicited an LMI response with cells from healthy EBV-seropositive individuals whereas the extract from the parental DG75 cell line did not. The results demonstrate the value of stably transfected cell lines expressing a defined EBV antigen for the monospecific analysis of host responses to the EBV-encoded antigen complex in growth-transformed cells. PMID- 3040605 TI - Long term use of enalapril in the treatment of patients with congestive heart failure. AB - Fifty-five patients of both sexes, aged 39-72, suffering from congestive heart failure of varying aetiology were admitted to a double blind study in which enalapril given in a dose of 5-10 mg twice daily or placebo was added to existing medication. A significant increase in exercise performance (P less than 0.004) was seen in the group treated with enalapril, an improvement which was still evident after 24 weeks of therapy. No significant difference was seen between the mean ejection fractions but subjective assessment suggested improvement in the enalapril-treated group. PMID- 3040606 TI - Nifedipine antagonizes alpha-adrenoceptor mediated splanchnic and systemic vasoconstriction in man. AB - Aim of the study was to investigate whether nifedipine interacts with alpha1 mediated splanchnic and systemic vasoconstriction in man. The effects of nifedipine on basal and phenylephrine (PE)-induced hemodynamic changes were studied in 6 normotensive men, using the hepatic venous catheter technique (indocyanine-green) and the thermodilution method. After a baseline period, nifedipine was given by constant infusion (0.5 microgram/kg/min) and, after equilibration, PE was infused at a constant rate in a dose sequence of 1, 2, 3, and 4 micrograms/kg/min, until the systolic blood pressure was raised by 30 mmHg. Control trials without nifedipine (saline) were also performed in each subject. At basal conditions nifedipine decreased systemic blood pressure and total systemic vascular resistance (TSVR) as well as total pulmonary vascular resistance (TPVR), and increased cardiac output (CO). PE provoked a dose dependent increase in systemic blood pressure, TSVR and TPVR, and decreased CO. These PE-effects were clearly attenuated by nifedipine. In the splanchnic vascular bed nifedipine increased estimated splanchnic blood flow (ESBF) and decreased splanchnic vascular resistance (SVR) at basal conditions. The PE induced decrease in ESBF and increase in SVR were inhibited or attenuated (depending on PE-dosage) by nifedipine. We conclude that nifedipine counteracts alpha1-adrenoceptor mediated systemic and splanchnic vasoconstriction and therefore, might also be useful in the treatment of intestinal vasospasm. PMID- 3040607 TI - Effects exerted by RU 41740 on oxidative metabolism and migration of rat polymorphonuclear leukocytes collected after induction of one acute non specific inflammatory reaction. AB - The activity of RU 41740, a glycoprotein extract from Klebsiella pneumoniae, endowed with immuno-modulating properties, has been investigated on polymorphonuclear (PMN) leukocyte functions. This report deals with the effect of RU 41740 on oxidative metabolism (assessed by chemiluminescence, 02 consumption and O2- production) and on chemotaxis and random migration (using agarose and Boyden chamber techniques). PMNs were collected from the rat pleural cavity after induction of one acute inflammatory reaction (pleurisy induced by injection of a suspension of calcium pyrophosphate crystals). Experiments were performed in parallel after in vivo treatment or incubation in vitro. RU 41740 enhanced PMN oxidative metabolism and inhibited PMN chemotaxis while random migration was only affected using agarose assay at high concentration. This effect on PMN migration was observed with at least two attractants. These observations have been obtained either after incubation in vitro or administration in vivo. The minimal effective dose was 1 mg/kg in vivo and 0.1 microgram/ml in vitro. These data suggest that RU 41740 acts directly on PMN membrane receptors. PMID- 3040608 TI - Restoration of Fc receptor-mediated desensitization of superoxide generation in human PMN by PMA. AB - The superoxide (O2-) generating activity of human polymorphonuclear leukocytes (PMN) was suppressed more than 50% compared with control cells by preincubation of the cells with soluble immune complexes (Fc receptor-mediated desensitization). The suppression was observed when wheat germ agglutinin (WGA) or N-formyl-L-methionyl-L-leucyl-phenylalanine (fMLP) was used as stimuli for O2- generation. However, the desensitized cells could produce as much O2- as control cells when they were stimulated with phorbol myristate acetate (PMA). The desensitized cells were pre-exposed to a small dose of PMA for a short period (2 3 min). By this procedure, the activity of the cells for WGA or fMLP induced O2- generation was almost completely restored. Similarly, the inhibitory effect of 3' deazaadenosing on WGA, fMLP or immune complexes (IC) induced O2- generation was abolished by the short-term pre-exposure of the cells to a small dose of PMA. PMID- 3040609 TI - Cyclic AMP and cyclic GMP phosphodiesterase activities in Hodgkin's disease lymphocytes. AB - Cyclic nucleotide phosphodiesterase (PDE) activities were studied in peripheral blood monocyte-depleted lymphocytes and enriched T-lymphocyte suspensions from thirteen patients with previously untreated Hodgkin's disease (HD) and fourteen age and sex matched healthy volunteers. Monocyte-depleted lymphocytes from HD patients showed PDE-activities which were two times higher than in their normal counterpart cells. The mean cAMP-PDE activity present in enriched HD T-lymphocyte suspensions was four times higher than in control T-lymphocytes, and the mean cGMP-PDE associated with HD T-lymphocytes was three times higher than in the controls. The hydrolytic activities present in both monocyte-depleted and T lymphocyte enriched cells suspensions remained unchanged in absence or in the presence of calmodulin and calcium. Since depressed cAMP and cGMP resting levels have been observed in HD lymphocytes and lymphocyte subpopulations, our results suggest that the elevated PDE activities are, at least in part, responsible for the alterations in lymphocyte cyclic nucleotide levels. PMID- 3040610 TI - Inhibition of angiotensin-converting enzyme by angiotensin I analogue peptide inhibitors. A kinetic study. AB - Angiotensin I analogues with a phosphonic acid group replacing the C-terminal carboxyl group were shown to be competitive inhibitors of angiotensin-converting enzyme. This new class of inhibitors was used to study the binding requirements of the angiotensin I-like ligands to the enzyme's active site. These studies indicate that angiotensin-converting enzyme recognizes at least five amino acid residues at the C-terminus of the peptide. The effect of pH on the binding of the most potent inhibitor peptide was compared to Captopril. The two inhibitors showed similar Ki-pH profiles despite their structural differences. Chloride enhanced the binding of the peptide inhibitor at both pH 9.0 and pH 6.5. At pH 9.0 the inhibitor peptide and the anion bind randomly to the enzyme, while at pH 6.5 the mechanism is ordered. In the latter case, the anion binds first to the enzyme. PMID- 3040611 TI - Failure to detect a DNA repair-related defect in the transfection of ataxia telangiectasia cells by enzymatically restricted plasmid. AB - We have transfected two SV40-transformed human fibroblast cell lines with plasmids in which double-strand breaks have been introduced by restriction enzymes, within or near the selected gene. Restriction of pSV2gpt with KpnI, as previously shown by Cox et al. (1986), reduced the frequency of transfection more in the ionizing radiation-sensitive ataxia-telangiectasia line AT5BIVA than in the resistant line MRC5V1. When the related plasmid pSV2neo was restricted with SmaI, the reduction in transfection was less in the ataxia-telangiectasia than in the normal cell line. Under our conditions, the apparent defect in transfection of AT5BIVA by pSV2gpt appears to be a result of the unusual sensitivity of the repair-deficient recipient to the selective agent. Loss of potential transfectants is exacerbated when transient gene expression is reduced by restriction of the plasmid. We suggest that a reduction in yield of transfectants with restricted plasmid in ataxia-telangiectasia cells cannot readily be used as evidence of a defect in DNA repair. Our results are also relevant to standard transfection experiments, since they emphasize the importance of optimizing selection when transient expression may be reduced, to ensure that potential transfectants are not killed by the selection regime. PMID- 3040612 TI - Opioid neuronal denervation in Gilles de la Tourette syndrome. AB - Increased striatal dopaminergic functions with heightened postsynaptic receptor sensitivity has been proposed to underlie the major clinical symptoms of Tourette's syndrome (TS). The beneficial response of the majority of TS patients to haloperidol supports the hyperdopaminergic pathophysiological concept of TS. However, in 5 recently encountered TS patients, haloperidol failed to ameliorate self-injurious behavior (SIB) while the opiate antagonist, naloxone, attenuated SIB, implicating deranged endorphinergic mechanisms in the pathophysiology of this disorder. Brain damage is commonly associated with partial neuronal denervation, denervation supersensitivity and neuronal habituation (Cannon's Law). While the motor tics of TS possibly reflect neuronal denervation of striatal dopaminergic neurons. SIB may represent opioid denervation with alterations in opioid receptor sensitivity possibly involving striato-limbic hypothalamic circuits. The effect of naloxone on SIB in TS could thus be explained on the basis of a modulatory effect of this drug on opioid receptor sensitivity. PMID- 3040613 TI - [Function of non-parenchymal cells of the liver]. PMID- 3040614 TI - [Liver cirrhosis as a risk factor in hepatocellular cancer]. PMID- 3040615 TI - The dichotomy between herpes simplex virus type 1-induced ocular pathology and systemic immunity. AB - Herpes simplex virus Type 1 (HSV-1) was injected into the mouse eye by the intravitreal (into the vitreous chamber; VC) or translimbal (across the limbus into the anterior chamber; TxL) route. These routes were compared for ocular pathology and systemic immunity. After VC inoculation, the virus-injected eye developed a viral infection, with the majority of opposite, uninjected eyes remaining intact and free of virus over the 4 week period of observation. In contrast, following translimbal inoculation, the entire virus-injected eye developed infection and inflammation together with subsequent chorioretinitis in the opposite uninjected eye. Systemic immunity induced by VC or TxL virus inoculation was similar to the effects of anterior chamber (AC) inoculation of the same dose of HSV-1: T cell-mediated DTH responses were suppressed while levels of anti-HSV neutralizing antibody were enhanced, compared to subcutaneously primed positive control mice. These findings demonstrate that HSV 1-induced ocular pathology does not necessarily correlate directly with systemic immunity. PMID- 3040616 TI - An immunogenetic analysis of resistance to herpes simplex virus retinitis in inbred strains of mice. AB - Specific inbred strains of mice have been shown to vary considerably in their resistance and susceptibility to herpes simplex virus (HSV) infection. We injected 2 X 10(5) plaque forming units (PFU) of the KOS strain of HSV-1 intracamerally into one eye of BALB/c, C57Bl/6, and F1 (BALB/c X C57Bl/6) mice. HSV-1 antigens were localized in frozen sections of enucleated eyes at 10 to 14 days post-inoculation. Injected eyes of BALB/c mice showed an anterior uveitis with HSV-1 antigens in the anterior segment and an intact retina free of HSV antigens. The retina of the contralateral uninjected eye was necrotic and contained HSV-1 antigens. In both C57Bl/6 and F1 mice, HSV antigens were limited to anterior segment structures in the injected eye, whereas, in contrast to BALB/c mice, the contralateral retina appeared histologically normal and contained no viral antigens. The C57Bl/6 and F1 strains remained relatively resistant to retinal infection even if pretreated with up to 800 Rads of irradiation. The retinas of normal or sublethally irradiated C57Bl/6 and F1, but not BALB/c strains, were also resistant to intravitreal injection of HSV. These results suggest that resistance to HSV retinitis is a dominantly inherited trait, which depends only partly upon immunologic factors and may be heavily influenced by the inherent ability of host cells from different murine strains to support a productive viral infection. PMID- 3040617 TI - Modification of relaxation of lipid protons by molecular oxygen and nitroxides. AB - Measurement of proton 1/T1 in model lipids from 0.00023 to 1.3 Tesla, at 5 degrees C and 37 degrees C, shows that both oxygen and nitroxides effectively enhance the relaxation rates of the protons of lipids. Equilibration with 1 atm of oxygen has a sixfold greater effect on lipid protons than on water protons, primarily because of the high lipid solubility of oxygen. The maximum relative relaxivity of oxygen for lipid protons occurs at 0.12-0.8 Tesla. Lipid soluble nitroxides have a four- to eightfold greater effect on lipid protons than on water protons. This high relaxivity, combined with the high lipid solubility possible with nitroxides, could lead to significant contrast in vivo in lipid environments. PMID- 3040618 TI - Proton relaxation in experimental clots varies with method of preparation. PMID- 3040619 TI - Antibodies to Epstein-Barr virus in patients with nasopharyngeal carcinoma in Northern Ireland. PMID- 3040620 TI - Cytomegalovirus mononucleosis--a report of 70 cases in a community hospital. AB - Seventy consecutive cases of cytomegalovirus mononucleosis (CMV-MN) were examined retrospectively from their clinical, epidemiological and serological aspects. The mononucleosis syndrome was complicated by various specific organ involvement in 15 (21%) of the patients. Serial serum samples were examined for specific CMV IgG, IgA and IgM antibodies by either the immunoperoxidase or enzyme-linked immunoassay techniques, or both. Specific anti-CMV IgM antibodies were found in 57 patients whose blood samples were collected for the first time at the second week of illness. In seven patients, CMV infection was diagnosed by a rise of specific IgG antibodies and the presence of specific IgA antibodies. These seven patients were generally older (mean age 68 years); four of them had a malignant disease and probably represent cases of recurrent CMV infection. An association between malignancy and recurrent CMV infection in the older age-group is suggested. Prompt diagnosis of CMV infection may exclude CMV-MN from the list of fevers of unknown origin, thus avoiding laborious patient workups. PMID- 3040622 TI - Endogenous digoxin-like material is of adrenal origin. PMID- 3040621 TI - Nonclassical congenital adrenal hyperplasia. PMID- 3040623 TI - Acute pancreatitis associated with rising cytomegalovirus titer. PMID- 3040624 TI - Salivary duct carcinoma--a clinicopathologic study of 12 cases. AB - Salivary duct carcinoma (SDC), a recently defined malignant tumor usually of major salivary glands, has probably been included in the group of adenocarcinomas, NOS. As yet, only a few descriptions of its clinical behavior have appeared. We have found 12 cases of SDC treated at our institution since 1970 and have reviewed their presentation and course. Despite total parotidectomy in most cases and radiotherapy in all, most patients have succumbed to their tumors, six with distant metastases. SDC appears to be a highly malignant tumor requiring aggressive combined therapy for locoregional control. The high incidence of systemic spread indicates a need for effective chemotherapy on an adjuvant basis. PMID- 3040625 TI - Glomus tumor: diagnosis and management. PMID- 3040626 TI - Base of tongue salivary gland tumors. AB - Salivary tumors of the base of the tongue are encountered infrequently. A retrospective review of medical records from 1955 to 1985 was undertaken to determine the incidence of occurrence and to assess the outcome of the therapy provided. One hundred seventy-eight minor salivary gland tumors of the oral cavity and oropharynx were identified, of which 22 (13%) were located in the tongue base. All were malignant. The most common histologic type was mucoepidermoid carcinoma (10 patients), followed by adenocarcinoma (6 patients), and adenoid cystic carcinoma (6 patients). Thirteen patients were available for a mean follow-up of 5 years. Treatment was most often a combination of wide resection combined with postoperative radiation therapy. Ten patients (77%) are alive, one with persistent disease 8 years after diagnosis. Three patients died within 2 years of diagnosis, one with intercurrent disease. Improved control of disease in this series, when compared to previously reported series, is attributed to adequate surgical therapy and adjuvant radiotherapy. The deaths in our series occurred in patients who were unable to proceed with the recommended therapy. These unusual lesions require aggressive multimodality treatment for improved survival. PMID- 3040627 TI - The present state and perspectives of micronucleus assay in radiation protection. A review. AB - The occurrence of micronuclei proved to be a sensitive biological indicator of clastogenic effects of many chemical and physical agents. The possibility of using the micronucleus technique in radiation protection as an alternative to the traditional chromosomal analysis has recently been followed with increasing interest. This review outlines the main biological and practical aspects of the micronucleus assay and discusses its potential to serve as a rapid and reliable measure of radiation overexposures and hypersensitivities. PMID- 3040628 TI - The influence of Sn(IV) on the mean size of 99mTc(Sn)MDP constituents at neutral pH. AB - The effect of addition of Sn(IV), Mg(II) and Ca(II) on the mean size of the 99mTc containing constituents of a 99mTc(Sn)MDP reaction mixture was determined by a gel chromatographic batch procedure that does not disturb the equilibria in the mixture. PMID- 3040629 TI - Spectrophotometric analysis of irradiated spices. AB - Seven different spices (thyme, cinnamon, coriander, caraway, pimento, paprika, black pepper) were treated by gamma radiation at an absorbed dose of 10 kGy, and the effect on chemical quality was determined. The effects of this dose were assessed by spectrophotometric analysis of some water-soluble constituents of spices (carbohydrates; carbonyl compounds) and on the content of water-insoluble steam-volatile oils. The colour of paprika and the content of piperine in pepper held in different packaging materials were measured in unirradiated and irradiated samples as a function of storage time. In all cases irradiation does not bring about any distinct qualitative or quantitative chemical changes based on spectrophotometric analysis of spice extracts. PMID- 3040630 TI - Comparison of different chromatographic quality-control procedures to determine the radiochemical purity of common 99mTc-diagnostic agents. AB - Simple and rapid chromatographic systems were used to test the radiochemical purity of eleven 99mTc-diagnostic agents. Those systems were selected which gave no oxidation or other by-products. The tests were done with equipment normally available in Nuclear Medical Departments. PMID- 3040631 TI - Fast high-yield labelling and quality control of [123I]- and [131I]MIBG. PMID- 3040632 TI - Radiosynthesis of a selective dopamine D-1 receptor antagonist: R(+)-7-chloro-8 hydroxy-3-[11C]methyl-1-phenyl-2,3,4,5-tetrahydro-1H-3 -benzazepine ([11C]SCH 23390). AB - Carbon-11 labeled SCH 23390, a selective dopamine D-1 receptor antagonist (Fig. 1b), was prepared by N-alkylation of the nor-methyl precursor with [11C]iodomethane. The product was purified by semi-preparative HPLC and shown to be radiochemically pure at the end of synthesis. The synthesis was completed in approximately 22 min with an average radiochemical yield of 28% (based on [11C]iodomethane) and an average specific activity of approximately 1950 microCi/mumol calculated at the end of synthesis. PMID- 3040633 TI - A method for direct quantification of the amount of DTPA in 111In monoclonal antibody preparations. AB - The bicyclic anhydride of DTPA-1-14C (BADTPA-1-14C) was synthesized and reacted with an antibody to human melanoma associated antigen (MAA) and with one to human class II major histocompatibility complex antigen (HLA-DR). DTPA-1-C incorporation per mole of anti MAA at molar ratios of 1:1 to 200:1 ranged from 0.5 to 25, while immunoreactivity ranged from 49 to 9%. With antibody to HLA-DR, results were similar. Anti MAA, but not anti HLA-DR, demonstrated polymerization upon conjugation. BADTPA-1-14C provides a convenient and accurate method for measuring the amount of DTPA in monoclonal antibody preparations and its effect on immunoreactivity. PMID- 3040634 TI - Production of gallium-66, a short-lived, positron emitting radionuclide. AB - The excitation functions for the production of 66Ga and some other radionuclides produced by the 4He-particle bombardment of a natural copper target have been measured up to 31 MeV incident energy. The radiochemical separation of 66Ga from the irradiated copper target as well as from the other radionuclides formed is described. The yield of 66Ga at 18 MeV incident energy is 708 microCi/microAh at end of bombardment and the contamination of 67Ga is 0.25%. The intended use of the 66Ga produced is to label monoclonal antibodies for PET scanning. PMID- 3040635 TI - Boronation of antibodies with mercaptoundecahydro-closo-dodecaborate(2-) anion for potential use in boron neutron capture therapy. AB - The anionic polyhedral borane derivative, mercaptoundecahydro-closo dodecaborate(2-), has been evaluated as a boronating agent for antibodies. The objective of these studies was the selective delivery of boron to neoplasms for neutron capture therapy. Incubation of a large excess of this anion with the polyclonal antibody antithymocyte globulin (ATG) resulted in the incorporation of 9-13 mol of the anion per mol of antibody. The extent of boron incorporation into the protein was measured either by tritium-labeled B12H11SH2- or by direct boron determination with neutron activation analysis. The nature of the covalent linkage of the anion to the antibody appeared to involve the formation of a new disulfide bond by a thiol-disulfide exchange. The number of boron atoms incorporated into antibodies by this method appeared to be inadequate for neutron capture therapy. However, such boronated antibodies may have potential for the detection of molecules of biologic interest by means of electron energy loss spectroscopy. PMID- 3040636 TI - Measurements of induced radioactivity in electron- and photon-irradiated beef. AB - Samples of beef were irradiated with electrons of approx. 10- and 13.5-MeV energies or with 60Co gamma-ray photons (1.17 and 1.33 MeV). Induced radioactivity was measured with a gamma-ray spectrometer, consisting of a Ge(Li) detector and a multichannel analyzer. No induced radioactivity could be detected in the photon-irradiated samples; also for 10-MeV electrons the activity was below the detection limit. The irradiation by 13.5-MeV electrons, however, resulted in measurable radioactivity and the amount of 13N-activity was in agreement with previously calculated values. These measurements confirm previous conclusions that irradiation of food with electrons at 10 MeV or even at 13.5 MeV does not constitute any health risk due to radioactivity. Part of the induced radioactivity from 13.5-MeV electron irradiation is due to neutron capture, and the results suggest that several neutron sources contribute to the measured radioactivity. PMID- 3040638 TI - Raman spectroscopic evidence for Tc-oxo cores in Tc-HEDP complexes. PMID- 3040637 TI - 131I labeled diphosphonates for the palliative treatment of bone metastases--IV. Syntheses of benzylidenediphosphonates and their organ distribution in rats. AB - Palliative treatment of bone pain induced by disseminated bone metastases can be performed with osteotropic, beta(-)-emitting radionuclides. Newly developed 131I labeled benzylidenediphosphonic acid (BDP) derivatives show osteotropic characteristics which suggest that they might possibly be useful radiopharmaceuticals for that purpose. Six BDP derivatives were synthesized with H, OH or NH2 in the 4- and alpha-position. Syntheses were performed by the formal addition of 2 mol of phosphorous acid in the presence of PBr3 to 1 mol of the respective benzonitrile. Transformation of the 4-methoxy and 4-nitro substituents, which were stable during the diphosphonate formation, to 4-HO and 4 NH2 was achieved by hydrolytic ether cleavage in boiling HBr and catalytic hydrogenation with Pd/C, respectively. Transformation of the alpha-amino to alpha hydroxy group was achieved by the action of NaNO2 in HCl. 4 Hydroxybenzylidenediphosphonic acid (9) was formed unexpectedly during the reaction of 4-hydroxybenzoic acid with H3PO3/PBr3. The addition of 2 mol of phosphorous acid to the benzoic acid was accompanied by an additional reduction at the alpha-carbon. The new BDP derivatives were analyzed by HPLC, NMR and elemental analysis. After labeling with 131I the BDP derivatives were tested in female Sprague-Dawley rats to obtain organ uptake and kinetic data. The various substituents showed an influence on the bone affinity and the uptake in other organs. Among the BDP derivatives tested alpha-amino-(3-[131I]iodo-4 hydroxybenzylidene)diphosphonate (4a) showed the best biological characteristics. PMID- 3040639 TI - Preparation of thin self-supporting samples of liquids for external beam PIXE analyses: application to human cerebrospinal fluid. PMID- 3040640 TI - Accurate determination of 14CO2 by expulsion from blood. PMID- 3040641 TI - A nutritional analysis of food provided to Royal Naval personnel at sea. AB - The aim of this study was to determine the nutritional content of the foods provided to, and the confectionary purchased by, personnel serving at sea in British warships. Data were collected from the stores accounts of six ships over a period of 32,354 man-victualling-days. Analysis was carried out at the Nuffield Laboratories of Comparative Medicine. The food from the ships' gallies provided mean daily intakes of 3750 kcal, of which 42 per cent were derived from fats (17 per cent from saturated fats) and 9 per cent from added sugars. In taking account of confectionary purchased from NAAFI sources the total energy intake was raised to 4200 kcal of which 40 per cent were derived from fats (15 per cent saturated fats) and 12 per cent from added sugars. The results do not compare favourably with the recommendations of the reports of expert committees. PMID- 3040643 TI - [Acute viral infections in association with idiopathic peripheral facial paralysis]. AB - The possible association of some viral infections with the onset of Bell's palsy was examined in a study of 29 patients. The results were compared with a sex- and age-matched control group. The number of probable recent viral infections, as judged by a fourfold increase in antibody titers or the presence of specific IgM antibodies, differed statistically from that found in the control group. In seven patients with Bell's palsy the enzyme-linked-immunosorbent assay (ELISA) indicated an acute viral infection (herpes simplex 4; varicella zoster 2; cytomegalovirus 1). All these infections were due to viruses belonging to the herpesvirus group. Clinical evidence of herpesvirus infection was found in three cases (Herpetic eruption). The aetiological relationship between the virological findings and Bell's palsy is discussed. Reactivated herpes simplex virus and transient demyelination of the facial nerve could be one cause of an idiopathic facial palsy. PMID- 3040642 TI - Light and electron microscopical immunocytochemistry of 5'-nucleotidase in rat cerebellum. AB - 5'-Nucleotidase in nervous tissue has so far not been localised at the ultrastructural level using immunocytochemical techniques. We have now applied monoclonal antibodies and a polyclonal antiserum raised against this ecto-enzyme and describe the distribution of 5'-nucleotidase antigenicity in rat cerebellum both at the light and electron microscopic levels. Within all cerebellar layers, 5'-nucleotidase immunoreactivity was found on plasma membranes of glial elements, i.e. Bergmann glial cell processes crossing the molecular layer, astrocytic end feet around blood vessels and glial cell extensions surrounding single Purkinje cells. In the granular layer, 5'-nucleotidase immunoreactivity was present on glial membranes interposed between granule cells. Neuronal cells or processes were devoid of immunoreactivity. The immunocytochemical results were compared with conventional 5'-nucleotidase histochemistry. Both techniques showed the same ecto-localisation of the enzyme and favour the view of 5'-nucleotidase being predominantly situated at glial plasma membranes. PMID- 3040644 TI - Differential antigen presentation by heat-treated peripheral blood mononuclear cells and Epstein-Barr virus-transformed lymphoblastoid cell lines (EBV-LCL): heated EBV-LCL present alloantigen and soluble antigen but are deficient in the stimulation of autologous EBV-LCL primed T cells. AB - Heat-treated PBM (1 hr at 45 degrees C) cannot present soluble Candida albicans antigens (CAN) or stimulate in the mixed lymphocyte culture (MLC) reaction. This is despite their continued expression of serologically defined class II MHC antigens. In contrast, heat-treated EBV-LCL present soluble CAN and stimulate allogeneic T cells in the MLC. Heated EBV-LCL stimulate strong secondary responses from allogeneic alloprimed T-cell lines in the primed lymphocyte test (PLT), while heated PBM stimulate only weak secondary allogeneic responses. To test whether this difference was due to a subtle difference in the thermal stability of the functional expression of class II MHC antigens on PBM and EBV LCL cells, the EBV-LCL cells were heated for 1 hr at temperatures from 45 degrees C to 60 degrees C. Even after treatment at 60 degrees C, the heated EBV-LCL strongly stimulated alloreactive T cells in MLC and PLT reactions. Heated EBV-LCL are not nonspecifically mitogenic, as they do not stimulate autologous T-cell lines primed to alloantigens. However, the weak response of alloprimed T-cell lines to heated allogeneic PBM can be augmented by coculturing with autologous heated EBV-LCL, suggesting heated EBV-LCL maintain a metabolic activity necessary for allogeneic stimulation that is deficient in heated PBM. While heated EBV-LCL stimulate allogeneic alloprimed T-cell lines, they no longer stimulate autologous EBV-LCL primed T-cell lines; irradiated EBV-LCL stimulate both strongly. This suggests the involvement of a heat labile antigenic or metabolic factor in the T cell recognition of autologous but not allogenic EBV-LCL. PMID- 3040645 TI - Paget's disease of the nipple without detectable breast tumor: conservative management with radiation therapy. AB - Between 1960 and 1984, 20 selected patients with Paget's disease of the breast confined to the nipple were treated conservatively with radiotherapy alone (17/20 pts) or limited surgery and radiotherapy (3/20 pts). Median follow-up was 7.5 years. No patients died of breast disease. Three patients had recurrence in the treated breast, and were treated by mastectomy. All recurrences were located in the nipple or areola and were all Paget's disease, without associated intraductal or invasive carcinoma. No axillary node recurrences occurred. The actuarial 7 year probability of living free of disease with breast preserved was 81%. Among the 15 patients who had a minimum follow-up of 3 years, without recurrence, 12 (80%) had a good cosmetic result. These results suggest that radiation therapy could be an effective alternative to radical surgery in the treatment of patients with Paget's disease of the nipple without concomitant breast tumor. PMID- 3040647 TI - WR 2721 modification of type II cell and endothelial cell function in mouse lung after single doses of radiation. AB - The ability of WR 2721 to protect endothelial cells and Type II cells in mouse lung after single doses of X rays was studied using specific assays of cell function to assess damage. The whole thorax of mice was exposed to a range of single doses of X rays either alone or 30 minutes after an i.p. injection of 400 mg/kg of WR 2721. Endothelial cell function was assayed by angiotensin converting enzyme (ACE) and Type II cell function by phosphatidylcholine and total protein present in lavage fluid 28 days after radiation. Similar protection factors (PFs) were found for the functional activity of both cell types, 1.2 and 1.24 for ACE and phosphatidylcholine respectively. These values were somewhat less than the PF of 1.37 for lethality from pneumonitis 7 to 9 months after irradiation for this mouse strain. The lack of a clear difference between the PFs for the functional activity of these two cell types suggests that neither the endothelial cell nor the Type II cell can be accepted or excluded as the target cell for radiation pneumonitis in lung. PMID- 3040646 TI - Functional responses of the pulmonary endothelium to thoracic irradiation in rats: differential modification by D-penicillamine. AB - Male rats were sacrificed 2 or 6 months after a range of single doses of gamma rays (0-30 Gy) to the right hemithorax. Half of each dose group consumed control feed continuously after irradiation, and half consumed feed containing the collagen antagonist D-penicillamine (10 mg/rat/day). Four markers of pulmonary endothelial function were monitored: angiotensin converting enzyme (ACE) activity, plasminogen activator (PLA) activity, and prostacyclin (PGI2) and thromboxane (TXA2) production. Bronchoalveolar lavage (BAL) fluid also was obtained from the right lung, and was analyzed for macrophage number, and PGI2 and TXA2 concentration. Right lung ACE and PLA activities decreased linearly with increasing dose at both 2 and 6 months postirradiation, and penicillamine had no significant effect on either response. In contrast, PGI2 and TXA2 production by the right lung increased linearly with increasing radiation dose at both autopsy times. Penicillamine significantly ameliorated the increase in PGI2 production at 2 months, and the increase in TXA2 production at both 2 and 6 months postirradiation. Penicillamine dose-reduction factors (DRF) for PGI2 and TXA2 production were 1.3-1.4, and the response curve slope ratios were 1.7-2.5 (p less than 0.05). Penicillamine also ameliorated the dose-dependent increase in TXA2 concentration in the BAL fluid at 2 months. These data indicate that the four "markers" of radiation-induced pulmonary endothelial dysfunction do not respond identically to penicillamine dose-modification. Of the four markers, TXA2 production exhibits the most significant and widespread penicillamine sparing. TXA2 is a potent vasoconstrictor, promoter of platelet aggregation, and mediator of inflammation, and partial prevention of the radiation-induced hyperproduction of this eicosanoid may account in part for penicillamine's therapeutic action in this model. PMID- 3040648 TI - Immune suppression in healthy carriers of adult T-cell leukemia retrovirus (HTLV I): impairment of T-cell control of Epstein-Barr virus-infected B-cells. AB - We have examined the activities of Epstein-Barr virus (EBV)-specific memory T cells of carriers of adult T-cell leukemia (ATL) associated retrovirus (HTLV-I) and non-carriers in an ATL-endemic area, all of whom were EBV-seropositive, by assay of EBV-induced B-cell focus regression. The result showed that immune suppression, as represented by lowering of the regression, was present in 18 (29%) out of 63 healthy HTLV-I carriers, in contrast to none of 63 matched control persons (healthy non-carriers). This finding suggests that a suppression of cell-mediated immunity is induced by a persistent infection with ATL retrovirus. PMID- 3040649 TI - Identification of transforming genes as hst in DNA samples from two human hepatocellular carcinomas. AB - In a DNA-mediated transfection assay using NIH3T3 cells, the DNAs from two out of twelve human hepatocellular carcinomas gave transformants. The transforming gene was identified as hst, which was originally found in DNAs from stomach cancers and a noncancerous portion of stomach mucosa, the putative product being a growth factor. Transcripts were not detected in the original HCC 1 and HCC2 upon Northern blot analysis. The possible mechanisms involved in the hst gene activation are discussed. PMID- 3040650 TI - A possible new member of tyrosine kinase family, human frt sequence, is highly conserved in vertebrates and located on human chromosome 13. AB - We have isolated a human genomic DNA (designated human frt) cross-hybridizing with the v-ros oncogene of UR2 sarcoma virus. Sequencing analysis of this fragment revealed that this sequence contains a 123-base-pair exon-like structure surrounded by consensus sequences of splice acceptor and donor sites. The deduced amino acid sequence of this stretch was highly homologous to a portion of a tyrosine kinase domain of src family. The human frt sequence was found to be conserved in a wide variety of vertebrates. By using a human-mouse hybridoma panel, human frt was located on chromosome 13, while human c-ros-1 was located on chromosome 6. PMID- 3040651 TI - The effect of 3-methylcholanthrene on postlanosterol cholesterol biosynthetic pathways in mouse skin. AB - Repeated topical application of 3-methylcholanthrene to the backs of BALB/c mice lowered the tissue levels of lathosterol and provitamin D3, intermediates in one of the cholesterol biosynthetic pathways (the delta 7 pathway), though the activities of lathosterol 5-desaturase and provitamin D3 7-reductase were similar to those of control animals. These results seemed to indicate that carcinogen treatment exerted a depressive effect at some earlier step(s) than lathosterol synthesis. However, the content of cholesterol in mouse skin was not lowered in these animals, suggesting that another biosynthetic pathway might be activated. When diazacholesterol, which is known to inhibit the conversion of desmosterol to cholesterol, was administered together with the carcinogen, a marked accumulation of desmosterol was observed compared to animals given only diazacholesterol. Since desmosterol is an intermediate in the pathway in which the delta 24 double bond is reduced at the final step (the delta 24 pathway), this seemed to suggest that the delta 24 pathway was activated by carcinogen treatment. PMID- 3040652 TI - Activation of N-ras gene in a rat hepatocellular carcinoma induced by dibutylnitrosamine and butylated hydroxytoluene. AB - DNA samples from eighteen rat hepatocellular carcinomas, including those induced by oral administration of dibutylnitrosamine (DBN) with butylated hydroxytoluene (BHT), or DBN with butylated hydroxyanisole (BHA), have been tested for the presence of transforming activity by transfection assay with NIH3T3 cells. Of the eighteen samples, only one from a tumor induced by DBN and BHT gave transformants. the activated oncogene was identified as rat N-ras by Southern blot analysis. PMID- 3040653 TI - Site-specific hypomethylation of the c-myc oncogene in human hepatocellular carcinoma. AB - In order to examine the effect of alteration in methylation of the c-myc gene on hepatocarcinogenesis, the extent of methylation of the c-myc gene was examined in 24 tissues of hepatocellular carcinoma (HCC), 24 adjacent non-tumor liver tissues from the same patients and 16 control liver tissues by the use of restriction endonucleases. The following results were obtained. (1) The c-myc gene from HCC tissue tended to be hypomethylated in comparison with that in non-tumor liver tissue from the same patient. (2) The c-myc gene from non-tumor liver tissue was hypomethylated to various degrees in comparison with that in control liver tissues. (3) The CCGG site in the third exon of the c-myc gene tended to be more extensively hypomethylated in HCC tissues than in non-tumor and control liver tissues. These results suggest that hypomethylation of the c-myc gene may occur to various degrees before the appearance of HCC, and may be associated with hepatocarcinogenesis. Moreover, the hypomethylation in the third exon of the c myc gene is probably important for the development of HCC. PMID- 3040654 TI - Appraising the instantaneous secretory rates of luteinizing hormone and testosterone in response to selective mu opiate receptor blockade in late pubertal boys. AB - The pulsatile properties of gonadotropin and testosterone release were examined before and after chronic mu opiate receptor blockade with naltrexone, 50 mg every other day, in four normal boys in late puberty (ages 14 8/12 to 15 1/12 years). The nature of spontaneous secretory events was appraised for immunoactive LH and testosterone in blood withdrawn every 20 minutes for 24 hours, using a novel, discrete deconvolution algorithm to estimate apparent instantaneous secretory rates. The application of this methodology revealed that the frequency of discrete LH instantaneous secretory rates increased after mu opiate receptor blockade (P = 0.011). More strikingly, all parameters of testosterone secretory events responded significantly to mu opiate receptor blockade, including increases in mean estimated secretory rate (+47%, P = 0.02), testosterone pulse frequency (+ 64%, P less than 0.001) and amplitude (+ 20%, P = 0.027). Correspondingly, decreases in testosterone interpulse secretory intervals (-35%, P = 0.001), secretory pulse duration (-19%, P = 0.042) and interpulse valley duration (-35%, P = 0.006) also were noted. There was a prominent diurnal rhythm in testosterone secretion with maximal values in the morning and late evening, and marked reductions in the afternoon, sometimes to prepubertal levels. This variation in the testosterone secretory profile paralleled that of LH. In response to naltrexone, the FSH concentration series showed a significant increase in the mean FSH concentration (+ 18%) P = 0.003) and mean peak amplitude (+ 15%, P = 0.002). These data provide indirect evidence of functional coupling of the opiate system with the hypothalamic GnRH pulse generator.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3040655 TI - Positive endocochlear potential: mechanism of production by marginal cells of stria vascularis. AB - The positive endocochlear potential (EP+) and high K+ concentration of the endolymph in the scala media of the mammalian cochlea are unusual. They have long been assumed to be due to a putative K-pump in the luminal membrane of the marginal cells of the stria vascularis, which were believed to have a negative internal potential. We show that the cell potential is more positive than the EP+, and that the ion pump is conventional Na,K-ATPase, probably in the basolateral membrane. The latter was determined from experiments in which the ionic environment of the strial cells was controlled by perfusion of the perilymphatic space of the cochlea, in the absence of vascular circulation. While the usual EP+ was maintained by normal perfusate, replacement of Na+ by choline resulted in a negative EP, showing that Na,K-ATPase is necessary for the production of EP+. Elimination of K+ as well as Na+ from the perfusate did not change the value of the negative EP, showing that no K-ATPase is involved. PMID- 3040656 TI - Activity of fiber-degrading microorganisms in the pig large intestine. AB - The large intestine is comparable to the rumen fermentation in many aspects; however, it is understood less well. Fiber in the form of cellulose and hemicellulose is one of the major substrates fermented in the large intestine. Various studies suggest that the pig can utilize fiber for growth, and up to 30% of its maintenance energy may be derived from volatile fatty acids produced in the large intestine. The total number of microorganisms in the pig large intestine do not change when a high fiber diet such as 50 or 80% alfalfa meal is fed. However, the fiber-degrading organisms increase and obviously replace others. The increase in fibrolytic bacteria normally coincides with an increase in enzyme activity (cellulase and xylanase), indicating that diet can be used to enhance fibrolytic activity. This is true for growing pigs and adult animals. The cellulolytic organisms in the pig, Bacteroides succinogenes and Ruminococcus flavefaciens, are similar to those in the rumen and are present at comparable numbers. This partly explains why adult pigs can maintain themselves by merely grazing on forage in pastures. Assuming other conditions are met, there is a significant potential for fiber degradation in the pig large intestine. Whether various genotypes such as the genetically selected obese and lean pigs have different abilities to degrade fiber is unknown. More work is required to understand the interaction of the fibrolytic organisms with the other organisms present in the large intestine, similar to that which has been done in the rumen, as well as the microbe-host interaction. PMID- 3040657 TI - Thoughts on fiber utilization in swine. AB - There is continued incentive for the development of alternative feed resources for use in swine production. The availability of distillery by-products may be expected to increase as the use of corn and cereal grains for ethanol production expands. The acceptability of distillery by-products, milling by-products, forages and other fibrous feeds as energy sources for swine depends on such factors as cell wall content of the plant, degree of microbial fermentation in the large intestine, and extent of absorption and utilization of the volatile fatty acids produced. Physiological effects of dietary fiber, including effects on gastric emptying, rate of transit of digesta, gut motility, digestive secretions, and absorption and utilization of breakdown products need quantification. Limited evidence suggests that there are genetic differences in the response of pigs to dietary fiber and in their ability to utilize it as an energy source. Recombinant DNA technology offers the possibility of cloning cellulase genes from microorganisms for application in swine feeding programs. The extent to which biotechnology will be applied in swine feeding will depend ultimately on the economic incentive for developmental effort and on unknown biological limitations of the pig and its gastrointestinal microbial ecosystem. PMID- 3040658 TI - Effects of bifonazole, fluconazole, itraconazole, and terbinafine on the chemiluminescence response of immune cells. AB - The luminol-enhanced chemiluminescence (CL) assay was used to examine the effects of antifungal agents tested at concentrations above and below therapeutically achievable levels on the CL response of mouse spleen cells. Reduction in the CL response of phagocytic cells may be indicative of an inhibition of the cellular immune response. Concomitantly, an increase in the CL response of phagocytic cells may indicate an enhancement of the immune capacity of these cells. The effects of four antifungal agents, bifonazole, fluconazole (UK-49,858), itraconazole, and terbinafine were studied. Changes in the CL response were assessed in terms of peak intensity, time to peak intensity, and area under the intensity-time curve compared with appropriate diluent controls for each drug. Both bifonazole and itraconazole caused significant reduction in peak CL intensity only at the highest level assayed (20 mg/l). Fluconazole had no significant effect on the CL response of mouse spleen cells at levels up to 20 mg/l, inclusive. Although terbinafine had no significant effect on peak CL intensity, it did cause a significant decrease in time to peak response at levels above 5 mg/l. This decrease in time to peak response may be indicative of an enhancement in the immune capacity of the mouse spleen cells; the clinical significance of this observation remains to be determined. PMID- 3040659 TI - Normal alveolar epithelial lining fluid contains high levels of glutathione. AB - The epithelial cells on the alveolar surface of the human lower respiratory tract are vulnerable to toxic oxidants derived from inhaled pollutants or inflammatory cells. Although these lung cells have intracellular antioxidants, these defenses may be insufficient to protect the epithelial surface against oxidants present at the alveolar surface. This study demonstrates that the epithelial lining fluid (ELF) of the lower respiratory tract contains large amounts of the sulfhydryl containing antioxidant glutathione (GSH). The total glutathione (the reduced form GSH and the disulfide GSSG) concentration of normal ELF was 140-fold higher than that in plasma of the same individuals, and 96% of the glutathione in ELF was in the reduced form. Compared with nonsmokers, cigarette smokers had 80% higher levels of ELF total glutathione, 98% of which was in the reduced form. Studies of cultured lung epithelial cells and fibroblasts demonstrated that these concentrations of reduced glutathione were sufficient to protect these cells against the burden of H2O2 in the range released by alveolar macrophages removed from the lower respiratory tract of nonsmokers and smokers, respectively, suggesting that the glutathione present in the alveolar ELF of normal individuals likely contributes to the protective screen against oxidants in the extracellular milieu of the lower respiratory tract. PMID- 3040660 TI - cAMP levels from endogenous and exogenous arachidonic acid in isolated dog lung. AB - We infused exogenous arachidonic acid (AA) into salt-perfused isolated dog lungs. This led to elevations in adenosine 3',5'-cyclic monophosphate (cAMP) which were from conversion of the AA to cyclooxygenase products. The maximal levels of cAMP occurred at far less than maximal levels of cyclooxygenase products. Next, we infused A 23187 to release endogenous pulmonary AA. This led to elevations in cAMP that were from conversion of this endogenous AA to cyclooxygenase products. The level of these products was far less than maximal levels from exogenous AA. However, maximal levels of cAMP from conversion of endogenous AA were similar to maximal levels of cAMP from conversion of exogenous AA. We conclude that maximal levels of pulmonary cAMP from endogenous or exogenous AA are from conversion of the AA to far less than maximal levels of pulmonary cyclooxygenase products. This indicates that levels of cAMP rather than levels of cyclooxygenase products are a potential rate-limiting step in cAMP-linked pulmonary actions of such products from pulmonary conversion of endogenous or exogenous AA. PMID- 3040661 TI - Liposome-mediated augmentation of catalase in alveolar type II cells protects against H2O2 injury. AB - Oxidant injury to the alveolar epithelium can be mediated by exposure to oxidant gases such as O2 at high concentrations and O3, inflammatory cell-derived reactive O2 species, and the intracellular metabolism of xenobiotics such as paraquat. An in vitro model of alveolar epithelial oxidant injury was developed based on exposure of cultured rat type II pneumocytes to superoxide and hydrogen peroxide (H2O2) enzymatically generated in the culture medium. Cytotoxicity was assessed by the release of lactate dehydrogenase (LDH) into the culture medium, which was a more reliable indicator of damage than release of 51Cr by prelabeled cells. Incubation of cells for 6-8 h with xanthine plus xanthine oxidase and glucose plus glucose oxidase induced the release of greater than 50% of total intracellular LDH. Oxidant exposure also resulted in significant detachment of cells from culture dishes. Modulation of oxidant damage was accomplished using liposomes as vectors for the delivery of catalase. Treatment of cells with catalase liposomes for 2 h resulted in augmentation of cellular catalase specific activities up to 631% of controls. Catalase was partitioned into intracellular and surface-associated compartments in catalase liposome-treated cells. Partial and complete protection against oxidant injury, induced by xanthine plus xanthine oxidase and glucose plus glucose oxidase, respectively, was achieved by pretreatment of cells with catalase liposomes. LDH release during oxidant exposure was inversely related to augmentation of cellular catalase activities. Catalase liposome-treated cells also exhibited an enhanced ability to scavenge enzymatically generated H2O2 from the culture medium. These observations suggest a useful approach to modulation of alveolar injury induced by reactive O2 species. PMID- 3040662 TI - Cloning and nucleotide sequence of the penicillinase antirepressor gene penJ of Bacillus licheniformis. AB - The penicillinase antirepressor gene, penJ, of Bacillus licheniformis ATCC 9945a was cloned in Escherichia coli by using pMB9 as a vector plasmid. The penicillinase gene, penP, its repressor gene, penI, and penJ were encoded on the cloned 5.2-kilobase HindIII fragment of the recombinant plasmid pTTE71. The penJ open reading frame was composed of 1,803 bases and 601 amino acid residues (molecular weight, 68,388). A Shine-Dalgarno sequence was found 7 bases upstream from the translation start site. Since this sequence was located in the 3' terminal region of the penI gene, penJ might be transcribed together with penI as a polycistronic mRNA from the penI promoter. Frameshift mutations of penJ were constructed in vitro from pTTE71, and the penJ mutant gene was introduced into B. licheniformis by chromosomal recombination. The transformant B. licheniformis U173 (penP+ penI+ penJ) turned out to be uninducible for penicillinase production, whereas the wild-type strain (penP+ penI+ penJ+) was inducible. Only when these three genes (penP, penI, and PenJ) were simultaneously subcloned in Bacillus subtilis did the plasmid carrier exhibit inducible penicillinase production, as did wild-type B. licheniformis. It was concluded that penJ is involved in the penicillinase induction. The regulation of penP expression by penI and penJ is discussed. PMID- 3040663 TI - A second regulatory gene, blaR1, encoding a potential penicillin-binding protein required for induction of beta-lactamase in Bacillus licheniformis. AB - A second regulatory locus (blaR1) required for the induction of beta-lactamase synthesis in Bacillus licheniformis 749 was cloned and sequenced. The gene was located on a 5.2-kilobase-pair SphI DNA fragment which also contained the beta lactamase (blaP) and repressor (blaI) genes. Bacillus subtilis BD224 carrying these three genes synthesized beta-lactamase on exposure to cephalosporin C, whereas Escherichia coli HB101 carrying the genes did not show any detectable induction of the enzyme. An open reading frame of 1,803 bases was identified as the blaR1 gene by subcloning and DNA sequencing. The gene started 2 bases downstream of the termination codon of bla1 and was preceded by a putative Shine Dalgarno sequence (AAGGA) with a spacing of 5 bases. The deduced blaR1 product (601 amino acids) had a molecular weight of 68,425. Five transmembrane regions were predicted from the hydrophobicity profile. The region around Phe-Ala-Pro-Ala Ser-Thr-Tyr-Lys (amino acids 398 to 405), which appeared to be located outside the membrane, was homologous to the binding regions of penicillin-binding proteins, including the beta-lactamases. The segment of 22 amino acids from 400 to 421 showed more than 70% homology to the penicillin-binding region of PBP 2 of E. coli. The blaR1 gene encodes a potential penicillin receptor which is required for the induction of beta-lactamase in B. licheniformis 749. PMID- 3040664 TI - Diversity of Chlamydia trachomatis major outer membrane protein genes. AB - Genomic DNA libraries were constructed for Chlamydia trachomatis serovars B and C by using BamHI fragments, and recombinants that contained the major outer membrane protein (omp1) gene for each serovar were identified and sequenced. Comparisons between these gene sequences and the gene from serovar L2 demonstrated fewer base pair differences between serovars L2 and B than between L2 and C; this finding is consistent with the serologic and antigenic relationships among these serovars. The translated amino acid sequence for the major outer membrane proteins (MOMPs) contained the same number of amino acids for serovars L2 and B, whereas the serovar C MOMP contained three additional amino acids. The antigenic diversity of the chlamydial MOMP was reflected in four sequence-variable domains, and two of these domains were candidates for the putative type-specific antigenic determinant. The molecular basis of omp1 gene diversity among C. trachomatis serovars was observed to be clustered nucleotide substitutions for closely related serovars and insertions or deletions for distantly related serovars. PMID- 3040665 TI - Cloning and characterization of dnaA(Cs), a mutation which leads to overinitiation of DNA replication in Escherichia coli K-12. AB - The product of the dnaA gene is essential for the initiation of chromosomal DNA replication in Escherichia coli K-12. A cold-sensitive mutation, dnaA(Cs), was originally isolated as a putative intragenic suppressor of the temperature sensitivity of a dnaA46 mutant (G. Kellenberger-Gujer, A. J. Podhajska, and L. Caro, Mol. Gen. Genet. 162:9-16, 1978). The cold sensitivity of the dnaA(Cs) mutant was attributed to a loss of replication control resulting in overinitiation of DNA replication. We cloned and sequenced the dnaA gene from the dnaA(Cs) mutant and showed that it contains three point mutations in addition to the original dnaA46(Ts) mutation. The dnaA(Cs) mutation was dominant to the wild type allele. Overproduction of the DnaA(Cs) protein blocked cell growth. In contrast, overproduction of wild-type DnaA protein reduced the growth rate of cells but did not stop cell growth. Thus, the effect of elevated levels of the DnaA(Cs) protein was quite different from that of the wild-type protein under the same conditions. PMID- 3040666 TI - Sequence determination and comparison of the exfoliative toxin A and toxin B genes from Staphylococcus aureus. AB - The DNA encoding the exfoliative toxin A gene (eta) of Staphylococcus aureus was cloned into bacteriophage lambda gt11 and subsequently into plasmid pLI50 on a 1,391-base-pair DNA fragment of the chromosome. Exfoliative toxin A is expressed in the Escherichia coli genetic background, is similar in length to the toxin purified from culture medium, and is biologically active in an animal assay. The nucleotide sequence of the DNA fragment containing the gene was determined. The protein deduced from the nucleotide sequence is a polypeptide of 280 amino acids. The mature protein is 242 amino acids. The DNA sequence of the exfoliative toxin B gene was also determined. Corrections indicate that the amino acid sequence of exfoliative toxin B is in accord with chemical sequence data. PMID- 3040667 TI - Nucleotide sequence of the epidermolytic toxin A gene of Staphylococcus aureus. AB - The nucleotide sequence of the eta gene, which codes for the epidermolytic toxin serotype A of Staphylococcus aureus TC16, is reported. The coding sequence of 840 nucleotides specifies a protein which, when secreted, has a predicted molecular weight of 26,950. The sequence of eta and the deduced amino acid sequence of the toxin have been compared with those of epidermolytic toxin serotype B. The coding sequences have 52% identical residues, and the polypeptides have 40% identical residues. Amino acid residues have been conserved in the areas of the proteins which correspond to major hydrophobic domains, whereas the regions likely to specify antigenic determinants occur in hydrophilic sequences that have diverged. The level of expression of epidermolytic toxin A in S. aureus 8325-4 was shown to be dependent on the integrity of a regulatory gene called agr. PMID- 3040668 TI - Nucleotide sequence of the Salmonella typhimurium metR gene and the metR-metE control region. AB - The nucleotide sequence of the Salmonella typhimurium metR gene and the metR-metE control region is presented. The metR gene codes for a polypeptide of 276 amino acids with a calculated Mr of 30,991. The metR gene product produced in a minicell system was found to migrate with an apparent Mr of 34,000. The transcription start sites for the metR and metE genes were determined by mung bean nuclease mapping. The metR and metE genes are divergently transcribed, with only 25 base pairs separating the transcription start sites. The overlapping nature of the metR and metE promoters suggests that there may be common regulatory signals for the two genes. PMID- 3040669 TI - Structural and genetic analyses of a par locus that regulates plasmid partition in Bacillus subtilis. AB - The Bacillus plasmid pLS11 partitions faithfully during cell division. Using a partition-deficient plasmid vector, we randomly cloned DNA fragments of plasmid pLS11 and identified the locus that regulates plasmid partition (par) by cis complementation in Bacillus subtilis. The cloned par gene conferred upon the vector plasmid a high degree of segregational stability. The par locus was mapped to a 167-base-pair segment on pLS11, and its nucleotide sequence was determined. The cloned par fragment regulated the partition of several different Bacillus replicons, and it only functioned in cis; it did not contain the replication function nor elevate the plasmid copy number in B. subtilis. The expression of par was orientation specific with respect to the replication origin on the same plasmid. We propose that the pLS11-derived par functions as a single-stranded site that interacts with other components involved in plasmid partition during cell division. PMID- 3040670 TI - Comparison of dnaA nucleotide sequences of Escherichia coli, Salmonella typhimurium, and Serratia marcescens. AB - The dnaA genes of Salmonella typhimurium and Serratia marcescens, which complemented the temperature-sensitive dnaA46 mutation of Escherichia coli, were cloned and sequenced. They were very homologous to the dnaA gene of E. coli. The 63 N-terminal amino acids and the 333 C-terminal amino acids of the corresponding DnaA proteins were identical. The region in between, corresponding to 71 amino acids in E. coli, exhibited a number of changes. This variable region coincided with a nonhomologous region found in the comparison of E. coli dnaA and Bacillus subtilis "dnaA" genes. The regions upstream of the genes were also homologous. The ribosome-binding area, one of the promoters, the DnaA protein-binding site, and many GATC sites (Dam methyltransferase-recognition sequence) were conserved in these three enteric bacteria. PMID- 3040671 TI - Analysis of transfer genes and gene products within the traB-traC region of the Escherichia coli fertility factor, F. AB - A series of plasmids that carry overlapping segments of F DNA encoding the genes in the traB-traC interval was constructed, and a restriction enzyme map of the region was derived. Plasmids carrying deletions that had been introduced at an HpaI site within this interval were also isolated. The ability of these plasmids to complement transfer of F lac plasmids carrying mutations in traB, traV, and traW, and traC was analyzed. The protein products of the plasmids were labeled in UV-irradiated cells and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. These analyses showed that the product of traV is a polypeptide that migrates with an apparent molecular weight of 21,000. It was not detected when [35S]methionine was used to label plasmid products, but was readily detected in 14C-amino acid labeling experiments. A 21,500-dalton product appeared to stem from the region assigned to traP. A 9,000-dalton product was found to stem from a locus, named traR, that is located between traV and traC. No traW activity could be detected from the region of tra DNA examined. Our data also indicated that traC is located in a more promoter-proximal position than suggested on earlier maps. The plasmids constructed are expected to be useful in studies designed to identify the specific functions of the traB, -P, V, -R, and -C products. PMID- 3040672 TI - Comparative organization of nitrogen fixation-specific genes from Azotobacter vinelandii and Klebsiella pneumoniae: DNA sequence of the nifUSV genes. AB - In the facultative anaerobe Klebsiella pneumoniae 17 nitrogen fixation-specific genes (nif genes) have been identified. Homologs to 12 of these genes have now been isolated from the aerobic diazotroph Azotobacter vinelandii. Comparative studies have indicated that these diverse microorganisms share striking similarities in the genetic organization of their nif genes and in the primary structure of their individual nif gene products. In this study the complete nucleotide sequence of the nifUSV gene clusters from both K. pneumoniae and A. vinelandii were determined. These genes are identically organized on their respective genomes, and the individual genes and their products exhibit a high degree of interspecies sequence homology. PMID- 3040674 TI - Involvement of FtsZ protein in shift-up-induced division delay in Escherichia coli. AB - A nutritional shift-up from glucose minimal medium to LB broth was previously shown to cause a division delay of about 20 min in synchronized cultures of Escherichia coli, and a similar delay was observed after a nutritional pulse (a shift-up followed rapidly by a return to glucose minimal medium). Using synchronized cultures, we show here that the pulse-induced division delay does not require protein synthesis during the period in LB broth, suggesting that a nonprotein signal is generated by the shift-up and transmitted to the cell division machinery. The cell division protein FtsZ, target of the SOS-associated division inhibitor SfiA (or SulA), seems to be involved in the postshift division delay. Mutants in which the FtsZ-SfiA interaction is reduced, either sfiA (loss of SfiA) or ftsZ(SfiB) (modification of FtsZ), have a 50- to 60-min division delay after a shift-up. Furthermore, after a nutritional pulse, the ftsZ(SfiB) mutant had only a 10- to 16-min delay. These results suggest that the FtsZ protein is the target element of the cell division machinery to which the shift up signal is transmitted. PMID- 3040673 TI - Inhibition of exogenous 3-deoxy-D-manno-octulosonate incorporation into lipid A precursor of toluene-treated Salmonella typhimurium cells. AB - Analogs of 3-deoxy-D-manno-octulosonate (KDO) were designed to inhibit CTP:CMP KDO cytidylyltransferase (CMP-KDO synthetase). Since these analogs lacked whole cell antibacterial activity, a permeabilized-cell method was developed to measure intracellular compound activity directly. The method employed a mutant of Salmonella typhimurium defective in KDO-8-phosphate synthetase (kdsA), which accumulated lipid A precursor at 42 degrees C. Cells permeabilized with 1% toluene were used to evaluate inhibitor effect on [3H]KDO incorporation into preformed lipid A precursor. KDO incorporation proceeded through the enzymes CMP KDO synthetase and CMP-KDO:lipid A KDO transferase. Optimum KDO incorporation occurred between pH 8 and 9 and required CTP, prior lipid A precursor accumulation, and a functional kdsB gene product, CMP-KDO synthetase. The apparent Km for KDO in this coupled system at pH 7.6 was 1.38 mM. The reaction products isolated and characterized contained 1 and 2 KDO residues per lipid A precursor molecule. Several KDO analogs produced concentration-related reductions of KDO incorporation in toluenized cells with 50% inhibitory concentrations comparable to those obtained in purified CMP-KDO synthetase systems. Two compounds, 8-amino-2-deoxy-KDO (A-60478) and 8-aminomethyl-2-deoxy-KDO (A-60821), competitively inhibited KDO incorporation, displaying Kis of 4.2 microM for A 60478 and 2.5 microM for A-60821. These data indicated that the inactivity of the KDO analogs on intact bacteria was the result of poor permeation into cells rather than intracellular inactivation. PMID- 3040676 TI - Cloning of the gene for the surface array protein of Aeromonas salmonicida and evidence linking loss of expression with genetic deletion. AB - A gene bank of DNA from the fish pathogenic bacterium Aeromonas salmonicida was constructed in the bacteriophage lambda gt11. Phage lambda gt11/10G, a recombinant carrying a 4.0-kilobase fragment of A. salmonicida DNA, was found to express the surface array protein (A protein) in Escherichia coli. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the protein expressed from the cloned gene had a subunit molecular weight of 49,000, which was identical to that of subunits in the native assembled A layer. Genomic Southern analysis showed that the gene coding for this predominant cellular protein was in a single copy on the chromosome and was conserved among a wide range of A. salmonicida strains with different phenotypic characteristics and isolated from diverse geographic locations, fish species, and means of pathogenesis. Results of genomic blotting experiments also showed that loss of expression of the A layer resulting from growth at 30 degrees C was accompanied by genetic rearrangement in which N terminal sequences of the gene for A protein were lost by deletion. PMID- 3040675 TI - Identification of polypeptides encoded by an Escherichia coli locus (hflA) that governs the lysis-lysogeny decision of bacteriophage lambda. AB - We report the cloning of the Escherichia coli hflA locus, which governs stability of phage lambda cII protein and which has been proposed to encode or regulate a cII-specific protease. The hflA locus was cloned on an 18-kilobase DNA fragment by selecting for plasmids that carry the neighboring purA gene. The boundaries of hflA were delimited by analysis of deletions and insertions constructed in vitro and by use of transposon Tn1000. Maxicell analysis of the proteins encoded by the hflA-containing fragment shows that hflA consists of at least two nonoverlapping genes, hflC and hflK, encoding polypeptides of 37,000 (C) and 46,000 (K) daltons. We observe that insertions into one gene eliminate the corresponding polypeptide and greatly reduce synthesis of the other. We suggest that these two polypeptides (K and C) interact to form a multimeric complex and that free subunits are unstable. We have constructed two types of fusions between hflA and lacZ. One is an hflC-lacZ protein fusion constructed in vitro; the other is an hfl-lacZ operon fusion in which a Mu dX(Apr lac) has inserted into the hflK gene. We have used the operon fusion to infer the direction of transcription of the hflK gene- toward hflC and in the same direction as hflC. Last, we describe evidence that hflA contains an additional gene, hflX, encoding a 50,000-dalton polypeptide. PMID- 3040677 TI - Expression of a cloned Bacillus thuringiensis crystal protein gene in Escherichia coli. AB - The expression in Escherichia coli of a cloned crystal protein gene from Bacillus thuringiensis was investigated through the use of fusions of the crystal protein gene promoter to beta-galactosidase and catechol oxidase genes. Analysis of deletion and insertion derivatives of the crystal protein gene promoter showed that a region of B. thuringiensis DNA located between 87 and 258 base pairs upstream from the transcription initiation site caused reduced transcription from this promoter. Insertion of Tn5 145 base pairs upstream from the transcription initiation site resulted in overproduction of the crystal protein. S1 nuclease mapping experiments failed to detect transcription from an outwardly directed promoter in Tn5, indicating that the overproduction resulted from the disruption or repositioning of the transcription-suppressing region. PMID- 3040678 TI - Effects of temperature-sensitive variants of the Bacillus subtilis dnaB gene on the replication of a low-copy-number plasmid. AB - The dnaB gene of Bacillus subtilis is involved in the initiation of DNA replication and also in the binding of the chromosomal origin to the bacterial membrane. We studied the effect of temperature-sensitive dnaB mutants (dnaB1 and dnaB19) on the replication and on the DNA-membrane binding of the plasmid pKW1, which was derived from the low-copy-number plasmid pBS2. In the dnaB19 mutant, pKW1 was not able to replicate at the restrictive temperature. In the dnaB1 mutant, however, the dimeric form of pKW1 DNA was preferentially produced as the restrictive temperature, but the replication of the monomeric form was totally blocked. We also examined the effects of the dnaB(Ts) gene on the DNA-membrane binding of both the double-stranded and single-stranded DNA from pKW1. The single stranded DNA from pKW1 was prepared from the DNA of the phage M13 mp19, which contained the origin of replication of pKW1. In the dnaB1 mutant, pKW1 DNA in both the double-stranded and single-stranded form was released from the membrane at the restrictive temperature. On the other hand, in the dnaB19 mutant, only double-stranded DNA, and not single-stranded DNA, was released from the membrane at the restrictive temperature. These results suggest that the product of the dnaB gene has at least two domains which influence the replication of DNA and the binding of DNA to the cell membrane in separate ways. PMID- 3040679 TI - Molecular characterization of the Escherichia coli enterobactin cistron entF and coupled expression of entF and the fes gene. AB - The Escherichia coli entF gene, which encodes the serine-activating enzyme involved in enterobactin synthesis, has been localized to a 4.7-kilobase-pair DNA fragment inserted in the vector pBR328. This recombinant molecule, pITS32, restored the ability of an entF mutant to grow on low-iron medium and to produce enterobactin. Examination of its translation products by minicell and electrophoretic analyses revealed a protein of approximately 160,000 daltons, which we identified as the EntF protein. A small DNA segment from pITS32 containing the translational start site for entF allowed the low constitutive expression of beta-galactosidase when cloned (pITS301) upstream of the lacZ structural gene in the vector pMC1403. In contrast, a clone (pITS312) containing the identical entF-lacZ fusion and a larger region upstream of entF including the entire fes gene and extending into the fepA gene (whose transcription is in the opposite direction relative to entF) expressed beta-galactosidase in high yet inducible amounts in response to fluctuations in the metabolic iron concentration. Transposon insertion mutations in the fes gene but not an insertion near the 5' region of fepA in pITS312 reduced this high inducible expression to the low constitutive level seen for pITS301. These observations are most readily explained by the presence of a regulatory region located upstream of fes which mediates the iron-regulated expression of a transcript that includes the fes and entF genes. PMID- 3040680 TI - Cluster of genes controlling synthesis and activation of 2,3-dihydroxybenzoic acid in production of enterobactin in Escherichia coli. AB - The Escherichia coli gene cluster encoding enzymatic activities responsible for the synthesis and activation of 2,3-dihydroxybenzoic acid in the formation of the catechol siderophore enterobactin was localized to a 4.2-kilobase chromosomal DNA fragment. Analysis of various subclones and transposon insertion mutations confirmed the previously suggested gene order as entEBG(AC) and provided evidence to suggest that these genes are organized as three independent transcriptional units, composed of entE, entBG, and entAC, with the entBG mRNA transcribed in a clockwise direction. Plasmid-specific protein expression in E. coli minicells identified EntE and EntB as 58,000- and 32,500-dalton proteins, respectively, while no protein corresponding to EntG was detected. The EntA and EntC enzymatic activities could not be separated by genetic or molecular studies. A small DNA fragment encoding both activities expressed a single 26,000-dalton polypeptide, suggesting that this protein is a multifunctional enzyme catalyzing two nonsequential reactions in the biosynthetic pathway. A protein of approximately 15,000 daltons appears to be encoded by the chromosomal region adjacent to the entAC gene, but no known function in enterobactin biosynthesis or transport can yet be ascribed to this polypeptide. PMID- 3040681 TI - Plasmid transfer in Streptomyces lividans: identification of a kil-kor system associated with the transfer region of pIJ101. AB - The 8.9-kilobase Streptomyces plasmid pIJ101 is self-transmissible at high frequency into recipient strains. By genetic analysis of the transfer region of the plasmid, we identified six plasmid-encoded loci involved in gene transfer and the associated pocking phenomenon. Two loci, kilA and kilB, could not be cloned into Streptomyces lividans on a minimal pIJ101-based replicon unless suitable kil override (kor) genes were present, either in cis or in trans. korA could control the lethal effects of both kilA and kilB, whereas korB could control only the effects of kilB. KilB mutants were defective in their pocking reaction; kilA mutants produced no visible pocks whatsoever. Mutations in two other loci, tra and spd, produced no pocks and defective pocks, respectively. These results suggest that kilA, kilB, tra, and spd are intimately involved in plasmid transfer and that the actions of kilA and kilB are regulated by the products of the korA and korB genes. PMID- 3040682 TI - Developmental expression of three proteins from the first gene of the RNA polymerase sigma 43 operon of Bacillus subtilis. AB - The first gene of the Bacillus subtilis RNA polymerase sigma 43 operon, P23, has a protein-coding capacity of 23,000 daltons. Sequence analysis revealed three potential translational initiation sites within the same reading frame, which could encode proteins of 23,000 (P23), 19,000 (P19), and 9,000 (P9) daltons, respectively. An internal promoter (P3), which is expressed only during the sporulation stage, is located between the second and the third translational start sites. By protein fusion to the Escherichia coli beta-galactosidase gene, we showed that all three translational initiation sites of the P23 gene are used in vivo in both E. coli and B. subtilis, and regulation for differential expression of the three proteins during the development of B. subtilis is coupled to the transcriptional promoter switching mechanism. The physiological function of these multiple gene products is unknown and is currently under investigation. PMID- 3040683 TI - Cloning and characterization of the Escherichia coli K-12 alanine-valine transaminase (avtA) gene. AB - avtA, which encodes the alanine-valine transaminase, transaminase C, was cloned in vivo with high- and low-copy-number mini-Mu cloning vectors. The phenotype conferred by the cloned avtA+ gene usually depended upon the plasmid copy number; most high-copy-number avtA+ plasmids permitted isoleucine-requiring ilvE strains to grow in the absence of isoleucine (multicopy suppression), while low-copy number avtA+ plasmids did not. avtA was mapped to a 1.25-kilobase segment by comparison of the restriction maps of 24 independent mini-Mu plasmids and then by gamma-delta (Tn1000) mutagenesis of a pBR322-avtA+ plasmid. The direction of transcription of avtA on the cloned fragment was determined with fusions to a promoterless lac gene. PMID- 3040684 TI - Molecular cloning and sequencing of the hemD gene of Escherichia coli K-12 and preliminary data on the Uro operon. AB - DNA of plasmid pSAS1002TH (F' ilv+ hemD+ hemC+ cya+) was used to clone the hemD gene of Escherichia coli K-12. Due to poor transformability of the heme-deficient mutants, the restriction fragments of the F' plasmid were first cloned into a mobilizable derivative of pBR322, pSAS1211LP, which was then mobilized into a hemD recA mutant (E. coli SASX419AN). One recombinant plasmid, carrying a HindIII fragment of about 5 kilobases (kb), was shown to complement the hemD mutant and also a cya mutant of E. coli K-12, as well as a hemC mutant of Salmonella typhimurium LT2. Further subcloning of the insert enabled us to locate the hemD gene to a BamHI-PstI fragment (approximately 2.3 kb) which also carried the hemC gene. The hemD gene occupies a region close to the PstI end, since the deletion of a 0.6-kb fragment from this end resulted in loss of the ability to complement the hemD mutation. The use of the promoter-probe vector pK01 and the results of complementation showed that the hemD gene was transcribed under physiological conditions from the same promoter as the hemC gene, the direction of transcription being hemC-hemD. This allows us to define a new polycistronic operon of E. coli K-12, for which we propose the designation Uro operon. Sequencing of the hemD gene showed the presence of an open reading frame (ORF) of 738 nucleotides which could code for a protein with a molecular weight of 27,766, which should correspond to the hemD protein; the ORF starts with the last nucleotide of the hemC gene, the two genes having different reading frames. An ORF of at least 480 base pairs follows the hemD gene after a few nucleotides. The corresponding gene X, the function of which is unknown, might represent a third member of the Uro operon. PMID- 3040685 TI - Sequence analysis of the gtfB gene from Streptococcus mutans. AB - The nucleotide sequence of the gtfB gene from Streptococcus mutans GS-5, coding for glucosyltransferase I activity, was determined. The gene codes for a strongly hydrophilic protein with a molecular size of 165,800 daltons. The deduced amino acid sequence revealed a typical gram-positive bacterial signal sequence at the NH2 terminus of the protein and 3.5 direct repeating units (each containing 65 amino acids) at the COOH terminus. Nucleotide sequencing of the region immediately downstream from the gtfB gene revealed the presence of a putative gene coding for an extracellular protein. This open reading frame is partially homologous to the gtfB gene. PMID- 3040686 TI - Nucleotide sequence of a glucosyltransferase gene from Streptococcus sobrinus MFe28. AB - The complete nucleotide sequence was determined for the Streptococcus sobrinus MFe28 gtfI gene, which encodes a glucosyltransferase that produces an insoluble glucan product. A single open reading frame encodes a mature glucosyltransferase protein of 1,559 amino acids (Mr, 172,983) and a signal peptide of 38 amino acids. In the C-terminal one-third of the protein there are six repeating units containing 35 amino acids of partial homology and two repeating units containing 48 amino acids of complete homology. The functional role of these repeating units remains to be determined, although truncated forms of glucosyltransferase containing only the first two repeating units of partial homology maintained glucosyltransferase activity and the ability to bind glucan. Regions of homology with alpha-amylase and glycogen phosphorylase were identified in the glucosyltransferase protein and may represent regions involved in functionally similar domains. PMID- 3040687 TI - Cloning of the debranching-enzyme gene from Thermoanaerobium brockii into Escherichia coli and Bacillus subtilis. AB - The gene for an enzyme with single or dual specificity on complex carbohydrates has been transferred from its native host (Thermoanaerobium brockii), a thermophilic anaerobe, into Escherichia coli and Bacillus subtilis. Most of the gene coding region is in a 2.2-kilobase PstI fragment that is common to the E. coli and B. subtilis chimeric vectors pCPC902 and pCPC903, respectively. Although the T. brockii debranching enzyme secreted from B. subtilis was unglycosylated and had less thermostability, more enzyme was secreted from B. subtilis (0.80 to 1.0 U/ml) than from T. brockii (0.23 U/ml). E. coli did not export any measurable enzyme. From the fermentation broth of B. subtilis containing pCPC903, three active species of the debranching enzyme were separated; two species are possibly protease digestion products of the larger protein (105,000 molecular weight). Whereas the enzyme can cleave all of the alpha-1----6 glucosidic linkages (and none of the alpha-1----4 bonds) in pullulan, it hydrolyzed mostly alpha-1----4 and very few of the alpha-1----6 linkages in starch. Upon hydrolysis of pullulan by the enzyme, only maltotriose was produced, while starch was digested to various-sized oligomers. PMID- 3040688 TI - Characterization of a bacteriophage that carries the genes for production of Shiga-like toxin 1 in Escherichia coli. AB - The Shiga-like toxin 1-converting bacteriophage H-19B was recently shown to carry the structural genes for the toxin and was shown to have DNA sequence homology with phage lambda. We present evidence that the linear genome of bacteriophage H 19B has cohesive termini which become covalently associated during prophage integration. Integration occurs through a site on a 4-kilobase-pair EcoRI fragment located near the center of the bacteriophage chromosome. The relationship between bacteriophages H-19B and lambda was examined by Southern hybridization. Homologous regions were mapped on the respective chromosomes which corresponded to the regions of the J gene, the int-xis area, and the O and P genes of phage lambda. The H-19B tox genes were mapped to the right of the O and P gene homology, which was far away from the phage attachment site. We concluded that H-19B is a lambdoid bacteriophage. Unlike other toxin-converting bacteriophages, the toxin genes were not located adjacent to the phage attachment site. It appeared that the Shiga-like toxin 1 genes were not picked up by a simple imprecise prophage excision. H-19B could, however, have acquired chromosomally located toxin genes by a series of events involving deletion and duplication followed by aberrant excision. PMID- 3040689 TI - Nucleotide sequence and promoter mapping of the Escherichia coli Shiga-like toxin operon of bacteriophage H-19B. AB - We determined the nucleotide sequence of the Shiga-like toxin-1 (SLT-1) genes carried by the toxin-converting bacteriophage H-19B. Two open reading frames were identified; these were separated by 12 base pairs and encoded proteins of 315 (A subunit) and 89 (B subunit) amino acids. The predicted protein subunits had N terminal hydrophobic signal sequences of 22 and 20 amino acids, respectively. The predicted amino acid sequence of the B subunit was identical to that of the B subunit of Shiga toxin. The A chain of ricin was found to be significantly related to the predicted A1 fragment of the SLT-1 A subunit. S1 nuclease protection experiments showed that the two cistrons formed a single transcriptional unit, with the A subunit being proximal to the promoter. A probable promoter was identified by primer extension, and transcription was found to increase dramatically under conditions of iron starvation. A 21-base-pair sequence with dyad symmetry was found in the region of the SLT-1 -10 sequence, which was found to be 68% homologous to a region of dyad symmetry found in the 35 region of the promoter of the iucA gene on plasmid ColV-K30, which specifies the 74,000-dalton ferric-aerobactin receptor protein. Betley et al. (M. Betley, V. Miller, and J. Mekalanos, Annu. Rev. Microbiol. 40:577-605, 1986) have recently summarized evidence suggesting that the slt operon is under the control of the fur regulatory system. The area of dyad symmetry found in both promoters may represent a regulatory site. A rho-independent terminator sequence was found 230 base pairs downstream from the B cistron stop codon. PMID- 3040690 TI - Cloning of the ADPglucose pyrophosphorylase (glgC) and glycogen synthase (glgA) structural genes from Salmonella typhimurium LT2. AB - The structural genes of ADPglucose pyrophosphorylase (glgC) and glycogen synthase (glgA) from Salmonella typhimurium LT2 were cloned on a 5.8-kilobase-pair insert in the SalI site of pBR322. A single strand specific radioactive probe containing the N terminus of the Escherichia coli K-12 glgC gene in M13mp8 was used to hybridize against a S. typhimurium genomic library in lambda 1059. DNA from a plaque showing a positive hybridization signal was isolated, subcloned into pBR322, and transformed into E. coli K-12 RR1 and E. coli G6MD3 (a mutant with a deletion of the glg genes). Transformants were stained with iodine for the presence of glycogen. E. coli K-12 RR1 transformants stained dark brown, whereas G6MD3 transformants stained greenish yellow, and they both were shown to contain a 5.8-kilobase-pair insert in the SalI site of pBR322, designated pPL301. Enzyme assays of E. coli K-12 G6MD3 harboring pPL301 restored ADPglucose pyrophosphorylase and glycogen synthase activities. The specific activities of ADPglucose pyrophosphorylase and glycogen synthase in E. coli K-12 RR1(pPL301) were increased 6- to 7-fold and 13- to 15-fold, respectively. Immunological and kinetic studies showed that the expressed ADPglucose pyrophosphorylase activity in transformed E. coli K-12 G6MD3 cells was very similar to that of the wild-type enzyme. PMID- 3040692 TI - Characterization of the Erwinia carotovora pelB gene and its product pectate lyase. AB - The pelB gene encodes pectate lyase B, one of three pectate lyases identified in Erwinia carotovora EC. Pectate lyase B was purified from Escherichia coli containing the pelB gene on a recombinant plasmid. The activity of the protein was optimal at a pH of 8.3. The amino acid composition, N-terminal amino acid sequence, and C-terminal peptide sequence were determined and compared with the polypeptide sequence deduced from the DNA sequence of pelB. Purified pectate lyase B started at amino acid 23 of the predicted sequence, suggesting that a 22 amino-acid leader peptide had been removed. Pectate lyase B of E. carotovora EC and pectate lyase B of E. chrysanthemi EC16 contain 352 and 353 amino acids, respectively (N. T. Keen, S. Tanaki, W. Belser, D. Dahlbeck, and B. Staskawicz, J. Bacteriol. 168:595-606, 1986). The two proteins are 72% homologous on the basis of DNA sequence data, and 75% of the amino acids are identical. PMID- 3040691 TI - Biosynthesis of bacterial glycogen: primary structure of Salmonella typhimurium ADPglucose synthetase as deduced from the nucleotide sequence of the glgC gene. AB - The nucleotide sequence of a 1.4-kilobase-pair fragment containing the Salmonella typhimurium LT2 glgC gene coding for ADPglucose synthetase was determined. The glgC structural gene contains 1,293 base pairs, having a coding capacity of 431 amino acids. The amino acid sequence deduced from the nucleotide sequence shows that the molecular weight of ADPglucose synthetase is 45,580. Previous results of the total amino acid composition analysis and amino acid sequencing (M. Lehmann and J. Preiss, J. Bacteriol. 143:120-127, 1980) of the first 27 amino acids from the N terminus agree with that deduced from nucleotide sequencing data. Comparison of the Escherichia coli K-12 and S. typhimurium LT2 ADPglucose synthetase shows that there is 80% homology in their nucleotide sequence and 90% homology in their deduced amino acid sequence. Moreover, the amino acid residues of the putative allosteric sites for the physiological activator fructose bisphosphate (amino acid residue 39) and inhibitor AMP (amino acid residue 114) are identical between the two enzymes. There is also extensive homology in the putative ADPglucose binding site. In both E. coli K-12 and S. typhimurium LT2, the first base of the translational start ATG of glgA overlaps with the third base TAA stop codon of the glgC gene. PMID- 3040693 TI - Identification of the DNA sequence required for transposition immunity of the gamma delta sequence. AB - A plasmid containing the transposon gamma delta sequence was immune to further insertion of gamma delta (transposition immunity). Plasmids carrying a fragment containing either 0.2 kilobase pairs of the gamma end or 0.4 kilobase pairs of the delta end of the gamma delta sequence were immune, and other parts of the gamma delta sequence did not confer immunity. The terminal 38-base-pair (bp) sequence of the delta end of the gamma delta was sufficient to confer immunity, the 38-bp sequence of the gamma end conferred only moderate immunity, and the terminal 35-bp sequence, which was completely identical at both the gamma and delta ends, was insufficient to confer immunity. PMID- 3040694 TI - Efficient transformation of phytopathogenic strains of Xanthomonas species. AB - Efficient transformation and conjugation systems for use in phytopathogenic strains of Xanthomonas species were developed with a dual-function plasmid vector, pBXC12, which was constructed from a newly isolated Xanthomonas citri plasmid, pXCL6, and pBR328. By using this system, pBR328 could also transform the same strains of Xanthomonas. The systems make useful cloning vectors for the study of genes involved in the plant pathogenesis of this species. PMID- 3040695 TI - Physical map of the Rhodobacter sphaeroides bacteriophage phi RsG1 genome and location of the prophage on the host chromosome. AB - We constructed a physical map of the 50-kilobase-pair (kb) DNA of the temperate Rhodobacter sphaeroides bacteriophage phi RsG1, with the relative positions of the cleavage sites for the nine restriction endonucleases KpnI, HindIII, XbaI, ClaI, BclI, EcoRV, EcoRI, BglII, and BamHI indicated. Using biotinylated phi RsG1 DNA as a probe in hybridization studies, we detected homologies with virus DNA and fragments of restriction endonuclease-digested host chromosomal DNA but not with plasmid DNA. This indicates that the prophage is integrated into the host chromosome. In addition, the use of specific probes such as the 10.4-kb BglII A fragment and the 2.65-kb BamHI H fragment allowed the determination of the position of phage attachment site (attP). PMID- 3040696 TI - Reaction centers from Rhodopseudomonas sphaeroides in reconstituted phospholipid vesicles. I. Structural studies. AB - Reaction centers (RCs) from Rhodopseudomonas sphaeroides were reconstituted into asolectin vesicles by cosonication. Equilibrium centrifugation on sucrose gradients showed that the vesicles were homogeneous in density (i.e., lipid-to protein ratio) when reconstituted at a molar lipid-to-protein ratio between 500 to 1000. At lower ratios, a considerable fraction of RCs was not incorporated into closed vesicles, while at higher ratios, an increasing population of liposomes was protein-free. The average vesicle size decreased with increasing lipid-to-protein ratio, exhibiting considerable size heterogeneity within a sample. The average diameter of the largest and smallest population of vesicles, reconstituted at a molar lipid-to-protein ratio of 560, was 1200 and 400 nm, respectively. The orientation of reconstituted RCs with respect to the plane of the membrane was determined from the flash-induced rereduction kinetics of the special-pair bacteriochlorophyll dimer in the presence of reduced cytochrome c. The predominant orientation of RCs was such that the cytochrome c binding sites faced the external medium. The net orientation of RCs in reconstituted vesicles decreased with vesicle size and was strongly influenced by the ionic strength during reconstitution. PMID- 3040697 TI - Reaction centers from Rhodopseudomonas sphaeroides in reconstituted phospholipid vesicles. II. Light-induced proton translocation. AB - Unidirectional light-dependent proton translocation was demonstrated in a suspension of reconstituted reaction center (RC) vesicles supplemented with cytochrome c and 2,3-dimethoxy-5-methyl-1,4-benzoquinone (UQ0), a lipid- and water-soluble quinone. Proton translocation was detected only at alkaline pH. The pH dependence can be accounted for by the slow redox reaction between the reduced quinone (UQ0H2) and oxidized cytochrome c. This conclusion is based on (i) the pH dependence of partial reactions of the reconstituted proton translocation cycle, measured either optically or electrometrically and (ii) titration studies with cytochrome c and UQ0. At 250 and 25 microM UQ0 and cytochrome c, respectively, maximal proton translocation was observed at pH 9.6. This pH optimum can be extended to a more acidic pH by increasing the concentration of the soluble redox mediators in the reconstituted cyclic electron transfer chain. At the alkaline side of the pH optimum, proton translocation appears to be limited by electron transfer from the endogenous primary to the secondary quinone within the RCs. The light intensity limits the reconstituted proton pump at the optimal pH. The results are discussed in the context of a reaction scheme for the cyclic redox reactions and the associated proton translocation events. PMID- 3040698 TI - The phosphate-pyrophosphate exchange and hydrolytic reactions of the membrane bound pyrophosphatase of Rhodospirillum rubrum: effects of pH and divalent cations. AB - The relation that exist between the Pi-PPi exchange reaction and pyrophosphate hydrolysis by the membrane-bound pyrophosphatase of chromatophores of Rhodospirillum rubrum was studied. The two reactions have a markedly different requirement for pH. The optimal pH for hydrolysis was 6.5 Mg2+ or Pi for the enzyme; Mn2+ and Co2+ support the Pi-PPi exchange reaction partially (50%), but the reaction is slower than with Mg2+; other divalent cations like Zn2+ or Ca2+ do not support the exchange reaction. In the hydrolytic reaction, Zn2+, at low concentration, substitutes for Mg2+ as substrate, and Co2+ also substitutes in limited amount (50%). Other cations (Ca2+, Cu2+, Fe2+, etc.) do not act as substrates in complex with PPi. The Zn2+ at high concentrations inhibited the hydrolytic reaction, probably due to uncomplexed free Zn2+. In the presence of high concentration of substrate for the hydrolysis (Mg-PPi) the divalent cations are inhibitory in the following order: Zn2+ greater than Mn2+ greater than Ca2+ greater than or equal to Co2+ greater than Fe2+ greater than Cu2+ greater than Mg2+. The data in this work suggest that H+ and divalent cations in their free form induced changes in the kinetic properties of the enzyme. PMID- 3040700 TI - Interaction of horse cytochrome c with the photosynthetic reaction center of Rhodospirillum rubrum. AB - Mitochondrial cytochrome c (horse), which is a very efficient electron donor to bacterial photosynthetic reaction centers in vitro, binds to the reaction center of Rhodospirillum rubrum with an approximate dissociation constant of 0.3-0.5 microM at pH 8.2 and low ionic strength. The binding site for the reaction center is on the frontside of cytochrome c which is the side with the exposed heme edge, as revealed by differential chemical acetylation of lysines of free and reaction center-bound cytochrome c. In contrast, bacterial cytochrome c2 was found previously to bind to the detergent-solubilized reaction center through its backside, i.e., the side opposite to the heme cleft [Rieder, R., Wiemken, V., Bachofen, R., and Bosshard, H. R. (1985). Biochem. Biophys. Res. Commun. 128, 120 126]. Binding of mitochondrial cytochrome c but not of mitochondrial cytochrome c2 is strongly inhibited by low concentrations of poly-L-lysine. The results are difficult to reconcile with the existence of an electron transfer site on the backside of cytochrome c2. PMID- 3040699 TI - (Na+ + K+)-ATPase: on the number of the ATP sites of the functional unit. AB - Questions concerning the number of the ATP sites of the functional unit of (Na+ + K+)-ATPase (i.e., the sodium pump) have been at the center of the controversies on the mechanisms of the catalytic and transport functions of the enzyme. When the available data pertaining to the number of these sites are examined without any assumptions regarding the reaction mechanism, it is evident that although some relevant observations may be explained either by a single site or by multiple ATP sites, the remaining data dictate the existence of multiple sites on the functional unit. Also, while from much of the data it is clear that the multiple sites of the unit enzyme represent the interacting catalytic sites of an oligomer, it is not possible to rule out the existence of a distinct regulatory site for ATP in addition to the interacting catalytic sites. Regardless of the ultimate fate of the regulatory site, any realistic approach to the resolution of the kinetic mechanism of the sodium pump should include the consideration of the established site-site interactions of the oligomer. PMID- 3040701 TI - Interactions of the catabolite activator protein (CAP) at the galactose and lactose promoters of Escherichia coli probed by hydroxyl radical footprinting. The second CAP molecule which binds at gal and the one CAP at lac may act to stimulate transcription in the same way. AB - The catabolite activator protein (CAP) binding sites of the Escherichia coli galactose and lactose operons were probed by hydroxyl radical footprinting. This method reveals each base that is protected by the bound protein. The patterns of protection seen for the primary CAP sites at gal and lac were virtually identical. In the presence of RNA polymerase the footprint of the second CAP molecule at gal was found to be very similar to those at the other two sites. This upstream site in gal align's perfectly with the lac CAP site with respect to the start of P1 transcription. Replacing most of the gal second CAP site DNA with heterologous sequences did not abolish binding although it became noticeably weaker. In vitro transcription studies of this hybrid gal promoter DNA further demonstrated the reduced affinity of the second CAP. These results are consistent with molecular models proposed for specific CAP binding and suggest that the second CAP at gal may be responsible for overall stimulation of transcription at this operon. Thus, in spite of differences in stoichiometry, the mechanisms of activation by CAP at gal and lac may be quite similar. PMID- 3040702 TI - Calcium-independent phosphoinositide breakdown in rat basophilic leukemia cells. Evidence for an early rise in inositol 1,4,5-trisphosphate which precedes the rise in other inositol phosphates and in cytoplasmic calcium. AB - Aggregation of the receptor with high affinity for immunoglobulin E (IgE) in rat basophilic leukemia cells leads to a calcium-dependent and a calcium-independent hydrolysis of phosphoinositides. The increase in the levels of inositol phosphates induced in the absence of calcium is only 25% of that observed with 1 mM Ca2+. The inositol phosphates reach a new steady state level 2 min after stimulation in EGTA, whereas with calcium they continue to increase up to 15 min. A similar response is observed when the receptors are aggregated due to the interaction of bound IgE with antigen or with anit-IgE, or by the binding of IgE cross-linked chemically. The antigen-mediated response is inhibited by hapten and disruption of such antigen-antibody aggregates late after stimulation leads to a rapid decline in the levels of the inositol phosphates to basal values. Separation of the inositol phosphates by Dowex columns shows that there is a fast rise in inositol trisphosphate which peaks at 15 s and slowly declines to a lower plateau within 2 min. Analysis by high pressure liquid chromatography reveals a 5 fold increase in the levels of inositol 1,4,5-trisphosphate in less than 10 s after stimulation, which precedes any major change in the other inositol phosphates. Aggregation of the receptor in the absence of external calcium induces a transient increase in cytoplasmic calcium which reaches a maximum of approximately 25 nM over basal levels after activation. The onset of the rise in Ca2+ lags after the initial rise in the inositol 1,4,5-trisphosphate. PMID- 3040703 TI - The kinetics of phosphoinositide hydrolysis in rat basophilic leukemia (RBL-2H3) cells varies with the type of IgE receptor cross-linking agent used. AB - We have re-examined, by high pressure liquid chromatographic (HPLC) procedures, the hydrolysis of 3H-labeled inositol phospholipids in rat basophilic leukemia (RBL-2H3) cells. Previous studies showed no clear correlation between the release of any particular inositol metabolite and the calcium signal in these cells. Paradoxically no responses were observed when the cells were stimulated with the antigen, aggregated ovalbumin, in the absence of external Ca2+. We report here that in the absence of external Ca2+ aggregation of the IgE receptor by agents other than aggregated ovalbumin causes the release of small amounts of [3H]inositol phosphates and a small increase in levels of cytosol Ca2+ (approximately 25 nM). The response, however, varied with the type of stimulant used. Within seconds after addition of 24 mol of dinitrophenol conjugated with 1 mol of bovine serum albumin to cells primed with dinitrophenol-specific IgE there was a small burst in release of [3H]inositol 1,4,5-triphosphate, [3H]inositol 1,3,4,5-tetrakisphosphate, and [3H]inositol 1,3,4-trisphosphate which was followed by a gradual rise in inositol 1,3,4-trisphosphate, inositol bisphosphate, and inositol monophosphate. Eventually, all inositol phosphates reached different steady state levels which were maintained for at least 40 min. In contrast, the initial response to oligomeric IgE, which aggregates receptors at a relatively slow rate, was muted although the subsequent development of the response was the same. The levels of inositol pentakisphosphate and hexakisphosphate remained unchanged. These and other studies with cell extracts support the conclusion that inositol 1,4,5-trisphosphate, a putative messenger for release of intracellular Ca2+, was converted to inositol 1,3,4,5 tetrakisphosphate and thence to inositol 1,3,4-trisphosphate. Both trisphosphate metabolites were dephosphorylated in sequential fashion by phosphatase enzymes in the cytosolic and membrane fractions. However, the appearance of several isomers of inositol monophosphates and bisphosphates suggested that degradation proceeded through multiple pathways in the intact cell. PMID- 3040704 TI - Multiple lipid interactions of the Sendai virus fusogenic protein. AB - The membrane topology of the envelope of Sendai virus was investigated using various radioactive photoactivable hydrophobic reagents: 3-(trifluoromethyl)-3-(m [125I]iodophenyl)diazirine and the two phospholipid analogues, 1-palmitoyl-2-(2 azido-4-nitro)benzoyl-sn -glycero-3- phospho[3H]choline and 1-myristoyl-2,12 amino-(4-N-3-nitro-1-azidophenyl)dodecanoyl-sn-glycero- 3-phospho[14C]choline. The hemagglutinin-neuraminidase glycoprotein and the fusogenic (F) glycoprotein were labeled by all three probes, confirming that these proteins are integral components of the viral envelope. The labeled F glycoprotein, composed of the two subunits F1 and F2, was cleaved in situ with trypsin to yield two fragments, F32 (32 kDa) and F19 (19 kDa). F2 was not labeled by any of the probes, suggesting an external location; whereas F19 was labeled by all probes and hence contains the portion of the F glycoprotein which traverses the viral envelope. Fragment F32 reacted both with 3-(trifluoromethyl)-3-(m-[125I]iodophenyl)diazirine and with 1 palmitoyl-2-(2-azido-4-nitro)benzoyl-sn-glycero-3-phospho[3H]choline, but not with 1-myristoyl-2,12-amino-(4-N-3-nitro-1-azidophenyl)dodecanoyl-sn-glycero- 3- phospho[14C]choline. This result opens the possibility that the F glycoprotein is formed by a loop-like structure having multiple interactions with viral lipids. PMID- 3040705 TI - F protein-F protein interaction within the Sendai virus identified by native bonding or chemical cross-linking. AB - The spatial arrangement of the F protein spike in the Sendai virus was studied after purifying the protein and reconstituting it in lipid vesicles (Sechoy, O., Philippot, J. R., and Bienvenue, A. (1986) Biochim. Biophys. Acta 857, 1-12). The different components of the F protein spikes were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under various conditions of treatment, i.e. at different temperatures and sodium dodecyl sulfate concentrations, using different detergents for F protein solubilization (Triton X-100 and octyl glucoside), by fast protein liquid chromatography analysis, and by chemical cross linking between subunits with bifunctional agents such as dimethyl adipimidate and dithiobis(succinimidyl propionate). The F protein spike appeared to be a structurally stable complex, composed of a noncovalent association of four homooligomers, each consisting of two peptides, F1 and F2, linked by a disulfide bond. Octyl glucoside and Triton X-100 solubilized the F protein, preserving the tetramer, which is probably the native form. Using chemical cross-linking, a covalent bond was formed between two monomers. We hypothesize that the tetrameric form of the F protein in its native form (spike) consists of two identical dimers that can be chemically cross-linked in a stable complex. PMID- 3040706 TI - Characterization of vasoactive intestinal peptide receptors by a photoaffinity label. Site-specific modification of vasoactive intestinal peptide by derivatization of the receptor-bound peptide. AB - The biological effects of vasoactive intestinal peptide (VIP) are mediated by binding to a membrane-bound receptor. Probes designed to trap this receptor by binding to it in a covalent way may suffer from a greatly reduced affinity. We report here, for the VIP receptor, the use of a photoaffinity probe obtained by derivatization of receptor-bound VIP with para-azidophenylglyoxal. This method protected the parts of the molecule essential for receptor binding. The VIP derivative thus obtained became covalently linked when irradiated. In the dark, however, it exhibited normal VIP-like behavior and retained its biological activity. This derivatization method might be generally applicable when hormone analogues have to be prepared without loss of receptor affinity. Receptor characterization studies on liver plasma membranes showed the presence of high- and low-affinity binding sites with KD = 0.1 and 5 nM, respectively. Treatment of membranes with dithiothreitol causes loss of high-affinity binding. The high affinity site, trapped by the photoaffinity probe, resolved into two molecular mass forms, 50 and 200-250 kDa. Reduction of the receptor-probe complex left the 50-kDa form intact, whereas the amount of the 200-250-kDa form greatly diminished. We demonstrate the importance of the presence of disulfide bonds in one of the molecular forms involved in high-affinity binding. PMID- 3040707 TI - 5-Iminodaunomycin. An anthracycline with unique properties. AB - 5-Iminodaunomycin forms a 3:1 complex with Fe(III) at pH 7.4. Drug-metal complex formation is associated with a marked decline in absorbance at 548 and 593 nm and the appearance of a broad band above 625 nm. The 5-iminodaunomycin-Fe(III) complex reacts with hydrogen peroxide to yield .OH radicals. This reaction is at a maximum at a drug/iron ratio of 2:1, and the yield is far less than that obtained with the doxorubicin-iron complex. In contrast to the results with doxorubicin, the production of .OH declines markedly at high 5 iminodaunomycin/iron ratios. There is a close parallel between the formation of hydroxyl radicals and the ability of the 5-iminodaunomycin complex to nick supercoiled SV40 DNA. The suppression of both .OH and DNA damage at high 5 iminodaunomycin:iron ratios is the result of several factors. 1) The presence of DNA stimulates .OH production from the doxorubicin complex, but not 5 iminodaunomycin; 2) doxorubicin reduces its chelated Fe(III) to Fe(II), but 5 iminodaunomycin does not; 3) 5-iminodaunomycin forms such a stable drug-metal complex that solvent water and, therefore, presumably H2O2, has diminished access to the chelated iron. The affinity of 5-iminodaunomycin is such that it can quantitatively abstract iron from doxorubicin. As a result, 5-iminodaunomycin is an effective competitive inhibitor of .OH radical formation by the doxorubicin iron complex. PMID- 3040708 TI - Proton hyperfine resonance assignments in cyanide-ligated cytochrome c peroxidase using the nuclear Overhauser effect. AB - The development of the proton nuclear Overhauser effect (NOE) for hyperfine shifted resonances of cyanide-ligated cytochrome c peroxidase (Saccharomyces cerevisiae) has been studied. In the pre-steady state regime, the major effects are due to primary NOEs to nearest neighbor protons. This has been used to advantage in making assignments of all of the remaining unassigned, resolved, downfield hyperfine shifted resonances. This work also determined the relative orientation of the heme pyrrole II substituents which is the cis configuration with the 4 alpha-vinyl proton pointing away from the 3CH3. In addition to heme protons, resonances of histidine 175, threonine 180, and histidine 52 have been assigned. These results indicate some structural rearrangement of the distal amino acids accompanying ligation. PMID- 3040709 TI - Phytic acid. A natural antioxidant. AB - The catalysis by iron of radical formation and subsequent oxidative damage has been well documented. Although many iron-chelating agents potentiate reactive oxygen formation and lipid peroxidation, phytic acid (abundant in edible legumes, cereals, and seeds) forms an iron chelate which greatly accelerates Fe2+-mediated oxygen reduction yet blocks iron-driven hydroxyl radical generation and suppresses lipid peroxidation. Furthermore, high concentrations of phytic acid prevent browning and putrefaction of various fruits and vegetables by inhibiting polyphenol oxidase. These observations indicate an important antioxidant function for phytate in seeds during dormancy and suggest that phytate may be a substitute for presently employed preservatives, many of which pose potential health hazards. PMID- 3040710 TI - Cyclic AMP inhibits both nicotine-induced actin disassembly and catecholamine secretion from bovine adrenal chromaffin cells. AB - As part of our studies on the functional role of the cytoskeleton in exocytosis we have reported (Cheek, T.R., and Burgoyne, R.D. (1986) FEBS Lett. 207, 110-114) that a calcium-independent transient disassembly of cortical actin filaments occurs on activation of the chromaffin cell nicotinic receptor but not when the cell is exposed to 55 mM K+. In order to determine whether this actin disassembly is required, in conjunction with a rise in intracellular Ca2+, to elicit a maximum secretory response from these cells, we have examined the relationship between actin disassembly, the elevation in intracellular Ca2+, and secretion in detail. The results show that the dose dependence of nicotine-induced secretion and actin disassembly are essentially identical with maximal effects at a dose of nicotine that produced a submaximal rise in intracellular Ca2+. Intracellular cAMP, elevated by three independent means, did not inhibit 55 mM K+-induced secretion but inhibited nicotine-induced secretion. Forskolin inhibited actin disassembly while not affecting the rise in intracellular Ca2+. These results demonstrate that a close inter-relationship exists between the secretory response and actin disassembly and provide further evidence suggesting that actin disassembly could be required in addition to the rise in intracellular Ca2+ in order to elicit a maximal secretory response in chromaffin cells. In addition, the results point to a role for cAMP in the regulation of stimulus-induced actin disassembly. PMID- 3040711 TI - Recognition of 5' and 3' splice site sequences in pre-mRNA studied with a filter binding technique. AB - A nuclear extract from HeLa cells was fractionated by DEAE-Sepharose chromatography. The obtained fractions were assayed for binding to an RNA transcript carrying a splice site sequence of 9-16 nucleotides by a filter binding technique. The U1 RNA-rich small nuclear ribonucleoprotein (snRNP) fractions, which showed binding activities for both 5' and 3' splice site RNAs, were studied for the sequence specificity of their binding. Results indicate that the U1-rich snRNP fraction can recognize both 5' and 3' splice site sequences. The U1 RNP, which was highly purified from the snRNP fractions, bound to at least some 5' splice site sequences, but not to a consensus 3' splice site sequence. Therefore, purified U1 RNP can directly recognize a 5' splice site, but not a 3' splice site. The binding activity for the 5' splice sites was lost either by digestion with micrococcal nuclease or by digestion of the 5' end of U1 RNA with RNase H and a complementary oligodeoxynucleotide, indicating the involvement of U1 RNA. Involvement of a protein moiety as well in this binding was suggested by the loss of binding activity upon heating at 60 degrees C. The binding activity to a 3' splice site sequence was not sensitive to digestion by micrococcal nuclease and was removed by protein A-coupled anti-Sm antibody. This activity was found in sucrose gradient fractions of about 8 S. PMID- 3040713 TI - Rat liver 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase. Identification of essential sulfhydryl residues in the primary sequence of the enzyme. AB - The kinase and sugar phosphate exchange reactions of rat liver 6-phosphofructo-2 kinase/fructose-2,6-bisphosphatase were inactivated by treatment with 5'-p fluorosulfonylbenzoyladenosine or 8-azido-ATP, but activity could be restored by the addition of dithiothreitol. This inactivation was accompanied by incorporation of 5'-p-sulfonylbenzoyl[8-14C]adenosine into the enzyme that was not released by the addition of dithiothreitol. The lack of effect of ATP analogs on the ATP/ADP exchange or on bisphosphatase activity and reversal of their effects on the kinase and sugar phosphate reactions by dithiothreitol suggest that 1) they reacted with sulfhydryl groups important for sugar phosphate binding in the kinase reaction, and 2) the inactivation of the kinase by these analogs involves a specific reaction that is not related to their general mechanism of attacking nucleotide-binding sites. In addition, alkylation of the enzymes' sulfhydryls with iodoacetamide prevented inactivation by 5'-p fluorosulfonylbenzoyladenosine, suggesting that the same thiols were involved. o Iodosobenzoate inactivated the kinase and sugar phosphate exchange; the inactivation was reversed by dithiothreitol; but there was no effect on the bisphosphatase or nucleotide exchange, indicating that oxidation occurred at the same sulfhydryl that are associated with sugar phosphate binding. ATP or ADP, but not fructose 6-phosphate, protected these groups from modification by 5'-p fluorosulfonylbenzoyladenosine, 8-azido-ATP, and o-iodosobenzoate. ATP also induced dramatic changes in the circular dichroism spectrum of the enzyme, suggesting that adenine nucleotide protection of thiol groups resulted from changes in enzyme secondary structure. Analysis of cyanogen bromide fragments of 14C-carboxamidomethylated enzyme showed that all radioactivity was associated with cysteinyl residues in a single cyanogen bromide fragment. Three of these cysteinyl residues are clustered in a 38-residue region, which probably plays a role in maintaining the conformation of the kinase sugar phosphate-binding site. PMID- 3040714 TI - Channeling of alpha-D-glucose 6-phosphate in tumoral islet cells exposed to D galactose. AB - In human erythrocytes, in which the fractional turnover rate of glucose 6 phosphate is rather low, menadione increases to almost the same relative extent the oxidation of D-[U-14C]glucose and D-[U-14C]galactose. However, in pancreatic tumoral islet cells (RINm5F line), in which the fractional turnover rate of glucose 6-phosphate is considerably higher, menadione increases the oxidation of D-[1-14C]glucose but not that of D-[1-14C]galactose. These results suggest that alpha-D-glucose 6-phosphate generated from exogenous D-galactose is channeled preferentially into the glycolytic rather than pentose phosphate pathway. Such was no more the case, however, when the RINm5F cells were exposed simultaneously to both D-glucose and D-galactose. PMID- 3040712 TI - The anomalous properties of the glutamate dehydrogenase-NADPH-alpha-ketoglutarate complex are not ascribable to a carbonyl addition reaction. AB - The glutamate dehydrogenase-NADPH-alpha-ketoglutarate complex, an active intermediate on the reaction pathway has a number of unusual properties: 1) it is the only blue-shifted natural complex of this enzyme; 2) it has an anomalously slow rate of dissociation; 3) its off-rate shows a substantial pH-independent D2O solvent isotope effect not exhibited by any other ternary complex of this enzyme; and 4) it has an unusually large enthalpy of interaction parameter. These properties must be ascribable to at least one of the two possibilities conferred on the complex by the presence of the alpha-carbonyl group of alpha ketoglutarate; the ability to engage in carbonyl addition reactions; and/or the ability to form a specific hydrogen bond. Oxalylglycine, a competitive inhibitor of alpha-ketoglutarate in this enzyme-catalyzed reaction, provides a means of discriminating between these two modes of action. The structure of oxalylglycine provides a dicarboxylic compound which has the same intercarboxylate proton distance and has a carbonyl group in a position spatially analogous to that of alpha-ketoglutarate. Its carbonyl group, however, is that of an amide group and cannot, therefore, engage in carbonyl addition reactions, but can hydrogen bond. Therefore, any effects observed with both oxalylglycine and alpha-ketoglutarate must be ascribed to formation of specific alpha-carbonyl hydrogen bonding, whereas any effects observed with alpha-ketoglutarate alone must be due to an alpha-carbonyl addition reaction. We have used this logic to test the source of the four phenomena listed above. In each case, oxalylglycine and alpha ketoglutarate showed the same effect. Therefore, we conclude that all four phenomena are in fact due to the formation of a specific alpha-carbonyl hydrogen bond and that the specific carbonyl addition reaction between alpha-ketoglutarate and an enzyme lysine group, postulated in one proposed catalytic mechanism, does not occur. PMID- 3040715 TI - The acceleration of Na+,K+-ATPase activity by ATP and ATP analogues. AB - Since Na+,K+-ATPase (EC 3.6.1.3) of pig kidney modified with a fluorescent sulfhydryl reagent, N-[p-(2-benzimidazolyl) phenyl]maleimide, at Cys-964 of the alpha-chain showed ATP-dependent, reversible, and dynamic fluorescence changes (Nagai, M., Taniguchi, K., Kangawa, K., Matsuo, S., Nakamura, S., and Iida, S. (1986) J. Biol. Chem. 261, 13197-13202), we studied the conformational change during Na+,K+-ATPase reaction using the modified enzyme. The addition of K+ to the enzyme increased the fluorescence intensity to 2% in the presence of 160 mM Na+ and 3 mM Mg2+ (K0.5 = 16.4 mM). Addition of low concentrations of ATP immediately increased the intensity to 3.2% (K0.5 less than 0.1 microM) to accumulate fully K+-bound enzyme in the presence of 43 mM K+ with Na+ and Mg2+, but further addition of higher concentrations of ATP diminished the increase (K0.5 = 120 microM). After exhaustion of ATP, the fluorescence intensity decreased to -0.4% (K0.5 = 0.3 microM) and -2% (K0.5 = 20 microM), respectively, in the presence of low and high concentrations of ADP produced from ATP. High concentrations of ATP accelerated Na+,K+-ATPase activity with a simultaneous increase in the amount of ADP-sensitive phosphoenzyme irrespective of the modification. Adenylyl imidodiphosphate and ADP accelerated Na+,K+-ATPase activity in the presence of 2.7 microM ATP by decreasing the extent of the fluorescence without affecting the amount of phosphoenzyme, irrespective of the modification. These data suggest that Na+,K+-ATPase activity was accelerated due to the acceleration of the breakdown of K+-bound enzyme by high concentrations of ATP and ATP analogues. PMID- 3040716 TI - Isolation and characterization of two types of cDNA for mitochondrial adenylate kinase and their expression in Escherichia coli. AB - Two cDNA clones for mitochondrial adenylate kinase were isolated from a cDNA library of bovine liver poly(A)+ RNA by using synthetic oligodeoxynucleotides as probes. The clone containing a 0.9-kilobase insert had the reading frame for a 241-residue protein (AK2A), while the other clone containing a 1.6-kilobase insert had the frame for a 234-residue protein (AK2B). Nucleotide sequences of these two clones were the same in the 5' portion up to the coding sequence for the 233rd residue, but different in the remaining 3' portions. The reported amino acid sequence of mitochondrial adenylate kinase from bovine heart corresponded to AK2A. Neither AK2A nor AK2B had a cleavable NH2-terminal presequence as that found in other imported mitochondrial proteins. RNA blot analysis of poly(A)+ RNAs from bovine liver and heart revealed three species of mRNA with approximate sizes of 0.9, 1.4, and 1.7 kilobases. The 1.7- and 1.4-kilobase species were specific for AK2B, whereas the 0.9-kilobase species was specific for AK2A. In the liver, the 1.7-kilobase mRNA was more abundant, whereas in the heart the 0.9 kilobase mRNA was predominant. The 1.4-kilobase mRNA was present only in the heart. The AK2A- and AK2B-coding sequences were expressed in Escherichia coli cells under the control of trc promoter. Both the products reverted the temperature-sensitive phenotype of the adenylate kinase mutant of E. coli. PMID- 3040717 TI - Primary structure and comparative sequence analysis of an insect apolipoprotein. Apolipophorin-III from Manduca sexta. AB - The amino acid sequence of an insect apolipoprotein, apolipophorin-III from Manduca sexta, was determined by a combination of cDNA and protein sequencing. The mature hemolymph protein consists of 166 amino acids. The cDNA also encodes for an amino-terminal extension of 23 amino acids which is not represented in the mature hemolymph protein. The existence of a precursor protein was confirmed by in vitro translation of fat body mRNA. Computer-assisted comparative sequence analysis revealed the following points: 1) the protein is composed of tandemly repeating tetradecapeptide units with a high potential for forming amphiphilic helical structures. Compared to mammalian apolipoproteins the repeat units in the insect apolipoprotein show considerable length variability; 2) the sequence has a striking resemblance to several human apolipoproteins including apoE, AIV, AI, and CI. However, the homology seems to be entirely functional since, although the insect and mammalian apoproteins contain very similar types of amino acid residues, the actual degree of sequence identity is quite low. Whether the mammalian and insect apoproteins are derived from a common ancestral amphiphilic helix forming, lipid-binding protein, or arose by convergent evolution can not be determined at present. This represents the first complete amino acid sequence for an insect apolipoprotein. PMID- 3040718 TI - Mutagenesis affecting the carboxyl terminus of the biotinyl subunit of transcarboxylase. Effects on biotination. AB - Biotin is added to biotin-containing enzymes as a post-translational modification catalyzed by holoenzyme synthetase. This reaction is fairly general in that synthetase from one organism will modify enzymes from heterologous sources. This suggests that the polypeptides share some structural characteristic(s) that define(s) them as biotin enzymes. We have reported previously that when the gene coding for the 1.3 S biotinyl subunit of transcarboxylase is expressed in Escherichia coli, the polypeptide produced is biotinated by the cellular synthetase. Using in vitro mutagenesis of this gene, we have begun to define the primary structure involved in the enzymatic addition of biotin to a lysine residue. We show here that the carboxyl terminus of the 1.3 S subunit is critical in biotination. Mutations affecting the COOH-terminal residue do not influence the modification, but elimination of the hydrophobic side chain of the penultimate residue abolishes biotin addition. PMID- 3040719 TI - Two asialoglycoprotein receptor polypeptides in human hepatoma cells. AB - Two cDNA clones isolated from a HepG2 lambda gt11 library encode the classical asialoglycoprotein receptor, H1, as well as a homologous membrane glycoprotein, H2 (Spiess, M., and Lodish, H.F. (1985) Proc. Natl. Acad. Sci. U.S.A. 82, 6465 6469). To study the relationship of H2 to H1 and its possible role in receptor mediated endocytosis of desialyated glycoproteins, we generated anti-peptide antibodies that are specific for each polypeptide. As judged by metabolic labeling of HepG2 cells and specific immunoadsorption, the biosynthesis of H2 is similar to H1 (Schwartz, A.L., and Rup, D. (1983) J. Biol. Chem. 258, 11249 11255); H2 is synthesized as a 43,000-dalton precursor polypeptide containing high mannose-type oligosaccharides, that is processed to a 50,000-dalton mature glycoprotein containing complex-type oligosaccharides. Both H1 and H2 have a half life of approximately 12 h. Trypsin and neuraminidase digestion of intact cells at 4 and 12 degrees C was used to determine that, at steady state, 50-60 percent of both H1 and H2 are on the cell surface. Furthermore, all of the H2 molecules were digested by extracellular neuraminidase in 1 h at 37 degrees C, indicating that all gain access to the plasma membrane. Both H1 and H2 were purified to homogeneity when Triton X-100-solubilized membrane proteins from [35S]cysteine labeled cells were subjected to affinity chromatography on galactose-agarose. Since we cannot detect a complex between mature H1 and H2, H2 must be a galactose binding protein. Both quantitative immunoprecipitation of each polypeptide from HepG2 cells and the recovery of purified H1 and H2 from galactose-agarose affinity chromatography indicate that there is 5-6 times more H1 relative to H2. That H2 is a minor species, compared to H1, might explain why it was not observed until a specific antibody was utilized. PMID- 3040720 TI - Translation initiation at an ACG triplet in mammalian cells. AB - The initiator AUC of the mouse dihydrofolate reductase gene (dhfr) was converted to ACG by site-directed mutagenesis and assayed for expression in cultured monkey cells using an SV40 recombinant called SVGT5dhfr26m2. Synthesis of apparently full-length dihydrofolate reductase (DHFR) protein was significantly reduced compared to wild-type, but not entirely abolished, suggesting that the ACG triplet was being utilized for translation initiation. In addition, a truncated form of DHFR was produced, apparently by initiation at the next in-frame AUG downstream. This result was confirmed in vitro. Transcripts of the dhfr sequence were produced by SP6 RNA polymerase in the presence of m7GpppG and translated in vitro using reticulocyte lysates and wheat germ extracts. The results paralleled those observed in vivo. Synthesis of full-length DHFR was reduced, but not eliminated, and a new species was produced by initiation at an internal site. Amino acid sequence analysis of the products of in vitro translation demonstrated that translation does indeed initiate at the ACG triplet and that it initiates with methionine. Additional mutations were introduced which altered the sequence context of the ACG triplet. Mutation of the translation initiation consensus sequence by substitution of the A residue at position -3, or of the G at +4 resulted in a significant decrease in initiation at the ACG and an increase in the level of the internal initiation product. Thus, translation initiation at a non-AUG triplet depends on a favorable sequence context. PMID- 3040721 TI - Agents that increase cAMP accumulation block endothelial c-sis induction by thrombin and transforming growth factor-beta. AB - Endothelial cells express the product of the c-sis gene, which encodes the B chain of platelet-derived growth factor (PDGF). Through local production of growth factors such as PDGF in vascular sites, endothelial cells may stimulate proliferation of adjacent cells through a paracrine mechanism. Previously, we have shown that the expression of c-sis mRNA and release of growth factor activity by human renal endothelial cells is induced by thrombin. We now show that another agent of possible importance in mediating proliferation of cells adjacent to the endothelial cell layer, transforming growth factor-beta (TGF beta), also induced c-sis expression in these cells. In addition, we have studied the effect of agents that increase intracellular cAMP levels upon the induction of endothelial cell c-sis mRNA. The adrenergic agonists isoproterenol and norepinephrine blocked the elevation of cellular c-sis mRNA accompanying exposure to either thrombin or TGF-beta. This effect was mediated through beta-adrenergic receptors, since propranolol but not phentolamine reversed the inhibition. Forskolin, a direct activator of adenylate cyclase, also blocked induction of c sis mRNA by thrombin and TGF-beta and inhibited the release of PDGF activity into the media of these cells. Basal, as well as stimulated c-sis mRNA levels were attenuated by these agents that increase cellular cAMP levels. These data suggest that increased cAMP production inhibits the expression of c-sis encoded mitogens by endothelial cells, and that c-sis expression is subject to bidirectional regulation in these cells. PMID- 3040722 TI - Identification of glutathione S-transferase Yb1 mRNA as the androgen-repressed mRNA by cDNA cloning and sequence analysis. AB - Androgens, while stimulating the growth of the rat ventral prostate, can also repress the levels of a limited number of mRNAs. The cDNA for one of the androgen repressed mRNAs has been identified by nucleotide sequence analysis as coding for the glutathione S-transferase Yb1 subunit. The prostate cDNA is 1071 nucleotides long, and only 2 or 4 bases of this sequence do not match the two published sequences of the cDNA for the Yb1 subunit of rat liver glutathione S-transferase. The amino acids in the protein encoded by the prostate cDNA matched completely with that for one of the liver cDNAs and differ with the other cDNA only in two of 218 amino acids. The identification of the androgen-repressed mRNA as a glutathione S-transferase subunit may indicate that some of the cellular actions of the enzyme may be important in the control of androgen-dependent growth of the prostate. Since Yb forms of the transferases have been colocalized with uridylic acid-rich small nuclear RNAs at interchromatinic regions of the cell nucleus, autoregulation of prostate growth by androgens may be carried out through the modulation of RNA production or processing in this target organ. PMID- 3040723 TI - Preferential repair of N-acetoxy-acetylaminofluorene lesions in the nuclease hypersensitive region of simian virus 40. AB - We exposed simian virus 40-infected CV-1 monkey cells to the carcinogen N-acetoxy acetylaminofluorene and monitored the removal of lesions from cellular DNA and from various regions of viral DNA. Exposure to 5.5 microM 3H-labeled carcinogen produced 20-80 adducts per 10(6) bases in cellular DNA in different experiments. The initial adduct concentration in viral DNA was always approximately half that in cellular DNA. At various times after treatment, cellular and viral DNA, and restriction fragments of viral DNA, were purified and examined for adduct density. Independent of the initial adduct concentration three rates of repair were observed. Cellular DNA was repaired at the lowest rate. Viral DNA was repaired about 50% more rapidly than was cellular DNA isolated from the same carcinogen-treated monolayers. Within the viral DNA a 366-base pair region containing the major nuclease-hypersensitive site was repaired at twice the rate of the rest of the viral genome. This region contains regulatory sequences that govern the initiation of DNA replication and viral gene expression. As reported previously this region was initially modified 1.71 +/- 0.20-fold higher than expected from its guanine content. Selective repair diminished the extent of hypermodification of this region by 6.0 +/- 2.1% per hour, partly compensating for the higher initial level of adducts. PMID- 3040724 TI - Characterization of nonheme iron and reaction mechanism of bromoperoxidase in Corallina pilulifera. AB - The properties of the nonheme iron of bromoperoxidase from Corallina pilulifera were studied. The enzyme lost its activity when reduced with formamidine-sulfinic acid and recovered it when oxidized by air. Incubation of the enzyme with ferric or ferrous ion-chelating agents indicated that its nonheme iron was ferric. Analyses of circular dichroism and proton NMR spectra suggested that the ferric ion tightly bound to cysteine, histidine, or tyrosine residues of the enzyme. The enzyme catalyzed Br--dependent catalase reactions to yield 1 mol of O2 from 2 mol of H2O2. No O2 evolution was observed when bromination reaction of monochlorodimedone occurred. From these results, together with previous knowledge of this enzyme, it was concluded that it activated bromide anion (Br-) to bromonium cation (Br+) using one molecule of H2O2, and this Br+OH- formed at the active site then decomposed another H2O2 to yield O2 in the absence of halogen acceptors (substrate). When substrate was present in the reaction mixture, it and H2O2 competitively reacted with the reaction intermediate (Br+OH-) to give brominated products. PMID- 3040725 TI - The reaction of superoxide, formate radical, and hydrated electron with transferrin and its model compound, Fe(III)-ethylenediamine-N,N'-bis[2-(2 hydroxyphenyl)acetic acid] as studied by pulse radiolysis. AB - Transferrin and the transferrin model compound Fe(III)-EHPG (Fe(III) ethylenediamine-N,N'-bis[2-(2-hydroxyphenyl)acetic acid] were found not to react with superoxide, as pulse radiolysis and kinetic spectroscopy revealed no transient species and no bleaching of the 465-nm absorption. However, transferrin was found to react with the formate radical, CO-.2, and the hydrated electron, e aq, with second-order rate constants of 3.8 X 10(8) and 1.1 X 10(10) M-1 S-1, respectively. These reactions produced a transient species (lambda max = 420 nm) which subsequently decayed by a second-order process. However, no reduction of the Fe(III) in transferrin was detected. Fe(III)-EHPG was also found to react with CO-.2 and e-aq, k = 7.3 X 10(6) and 1.1 X 10(9) M-1 S-1, respectively. The reactions of CO-.2 and e-aq with Fe(III)-EHPG resulted in no transient species but rather in reduction of the iron. These results are consistent with the inability of transferrin and Fe(III)-EHPG to catalyze the Haber-Weiss reaction. PMID- 3040726 TI - alpha 1-Antitrypsin nullGranite Falls, a nonexpressing alpha 1-antitrypsin gene associated with a frameshift to stop mutation in a coding exon. AB - alpha 1-Antitrypsin (alpha 1-AT) deficiency is a hereditary disorder associated with serum alpha 1-AT levels less than 35% of normal. There are two categories of alpha 1-AT genes that cause this state: the deficient alleles, in which alpha 1 AT is present in serum but in low levels, and the null alleles, in which no alpha 1-AT in serum can be attributed to the gene. The present study defines the molecular basis for the alpha 1-AT gene nullGranite Falls, identified and cloned from genomic DNA of an individual with severe alpha 1-AT deficiency and emphysema resulting from the heterozygous inheritance of the nullGranite Falls and Z alpha 1-AT genes. Sequencing of the 5'-flanking region, all five coding exons, and all exon-intron junctions of nullGranite Falls demonstrated it was identical with the common normal M1(Ala213) alpha 1-AT gene, except for two bases: a single deletion in the codon for amino acid Tyr160 of the mature protein and a single base substitution 168 base pairs 5' to exon I. Although no role for the promoter region mutation could be assigned, the coding exon deletion [Tyr(TAC)----(TA-)] resulted in a frameshift causing a stop coding to be formed approximately 44% from the N terminus of the precursor protein. Using oligonucleotide probes to evaluate the family of the index case demonstrated the deletion--- frameshift/stop mutation was inherited in an autosomal co-dominant fashion. Thus, although the molecular basis for alpha 1-AT deficiency of the alpha 1-AT null haplotype such as nullGranite Falls is very different from the molecular basis of the more common deficient haplotypes such as Z, the phenotypic consequences of the two genes are similar; i.e. severe alpha 1-AT deficiency and an association of a high risk for the development of emphysema. PMID- 3040727 TI - The onset of the respiratory burst in human neutrophils. Real-time studies of H2O2 formation reveal a rapid agonist-induced transduction process. AB - A real-time study of the initiation of the respiratory burst in human neutrophils was made. The cells were stimulated with fMet-Leu-Phe (fMLP) C5a, platelet activating factor, leukotriene B4, phorbol myristate acetate (PMA), or ionomycin, and H2O2 production was determined by chemiluminescence. Identical average onset times (2.4 s) and closely comparable values for the apparent first-order rate constant (kapp) for the induction of NADPH-oxidase activity (0.21-0.29 s-1) were obtained following stimulation with fMLP, C5a, platelet-activating factor, or leukotriene B4, suggesting that different agonists act through a common transduction sequence. Much longer onset times and lower kapp values were obtained upon stimulation with PMA or ionomycin. Pretreatment with PMA consistently shortened the onset time of the neutrophil's responses to agonists by about 1 s. When H2O2 production was initiated with PMA, a subsequent stimulation with the agonist fMLP elicited an immediate response (onset time less than 0.2 s) which preceded further changes in fura-2-detected [Ca2+]i. The results are consistent with a mechanism in which agonist signals appear to be transduced by two sequences acting in concert--a rate-limiting one liberating Ca2+ and diacylglycerol and turning on the Ca2+/phospholipid-dependent enzyme protein kinase C, and an essentially instantaneous one which does not appear to require further changes in cytosolic Ca2+. PMID- 3040728 TI - Analysis of ping-pong reaction mechanisms by positional isotope exchange. Application to galactose-1-phosphate uridyltransferase. AB - A new positional isotope exchange method has been developed that can be used for the analysis of enzyme-catalyzed reactions which have ping-pong kinetic mechanisms. The technique can be used to measure the relative rates of ligand dissociation from enzyme-product complexes. Enzyme is incubated with the labeled substrate and an excess of the corresponding unlabeled product. The partitioning of the enzyme-product complex back toward free enzyme is determined from the rate of positional isotope exchange within the original labeled substrate. The partitioning of the enzyme-product complex forward toward free enzyme is determined from the rate of formation of totally unlabeled substrate. It has been shown that the ratio of the two rates provides a lower limit for the release of product from the enzyme-product complex. The technique has been applied to the reaction catalyzed by galactose-1-phosphate uridyltransferase. The lower limit for the release of glucose 1-phosphate from the uridyl-enzyme relative to the maximal velocity of the reverse reaction was determined to be 3.4 +/- 0.5. PMID- 3040729 TI - Cloning, expression, and nucleotide sequence of a gene encoding a second thioredoxin from Corynebacterium nephridii. AB - A gene encoding thioredoxin in Corynebacterium nephridii was cloned in Escherichia coli by complementation of a thioredoxin mutant. Transformants that appeared to complement were analyzed for the presence of thioredoxin by the coupled assay using methionine sulfoxide reductase. Of 18 transformants, four contained high levels of thioredoxin activity. Transformants containing plasmids pLCN2 and pLCN4 were unable to support replication of T7 phage, in spite of their thioredoxin activities, and were studied in more detail. The plasmid pLCN2 contains a 1.85-kilobase Sau3AI insert, whereas pLCN4 contains a 10-kilobase TaqI insert. These plasmids complement all phenotypes of a thioredoxin-deficient strain except for replication of T7 phage. The nucleotide sequence of a 620-base pair HinfI fragment encoding thioredoxin derived from either plasmid indicated that the protein derived from this DNA is different from the thioredoxin of C. nephridii previously reported (Meng, M., and Hogenkamp, H.P.C. (1981) J. Biol. Chem. 256, 9174-9182). The amino acid sequence predicted from the nucleotide sequence shows a high degree of homology with other procaryotic thioredoxins. However, the new thioredoxin contains the tetrapeptide -Cys-Ala-Pro-Cys- at the active site and a third half-cystine residue in the carboxyl-terminal domain of the protein. The molecular weight of this thioredoxin, determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, is smaller than that estimated from the DNA sequence, suggesting that processing may have occurred. PMID- 3040730 TI - Characterization of inositol trisphosphate receptor binding in brain. Regulation by pH and calcium. AB - Inositol 1,4,5-trisphosphate is an intracellular second messenger, produced upon stimulation of the phosphoinositide system, capable of mobilizing calcium from intracellular stores. We have recently identified high levels of specific binding sites for inositol 1,4,5-trisphosphate in brain membranes (Worley, P. F., Baraban, J. M., Colvin, J. S., and Snyder, S. H. (1987) Nature 325, 159-161) and have now further characterized these sites. In cerebellar membranes, inositol 1,4,5-trisphosphate binding sites are abundant (20 pmol/mg protein) and display high affinity and selectivity for inositol 1,4,5-trisphosphate (KD approximately equal to 40 nM), whereas other inositol phosphates such as inositol 1,3,4,5 tetrakisphosphate (Ki approximately equal to 10 microM) and inositol 1,4 bisphosphate (Ki approximately equal to 10 microM) exhibit much lower affinity for this site. Submicromolar concentrations of calcium strongly inhibit inositol 1,4,5-trisphosphate binding (IC50 approximately equal to 300 nM). A sharp increase in binding occurs at slightly alkaline pH. These results suggest that actions of inositol 1,4,5-trisphosphate are regulated by physiological alterations in intracellular pH and calcium concentrations. PMID- 3040731 TI - The nitric oxide complex of ferrous soybean lipoxygenase-1. Substrate, pH, and ethanol effects on the active-site iron. AB - Soybean lipoxygenase is a non-heme iron enzyme that catalyzes the hydroperoxidation of linoleic acid by dioxygen. Exposure of ferrous lipoxygenase to nitric oxide yields a species displaying an electron paramagnetic resonance spectrum characteristic of a nearly axial S = 3/2 electronic spin system arising from the ferrous-nitrosyl complex. That spectrum is pH-sensitive, reflecting changes in the environment of the metal ion between pH 7 and 11. Addition of ethanol abolishes the effects of pH in a saturable fashion, resulting in a spectrum similar to that seen at pH 7. Exchange of lipoxygenase into H2(17)O leads to no significant line broadening in the low field portion of the spectrum, suggesting no coordination of water. The ferrous enzyme displays greater affinity for NO at pH 9 (where the enzyme is most active) than at pH 7. The binding of linoleic acid is competitive with that of NO at pH 9, but not at pH 7. These results are interpreted in terms of a model including only one iron site for exogenous ligands and an otherwise relatively stable iron coordination environment. PMID- 3040732 TI - Cyclic AMP responsiveness of human gonadotropin-alpha gene transcription is directed by a repeated 18-base pair enhancer. Alpha-promoter receptivity to the enhancer confers cell-preferential expression. AB - The human chorionic gonadotropin-alpha (CG-alpha) gene is transcriptionally activated by cAMP. Sequencing the CG-alpha 5'-flanking region identified two copies of a palindrome, 5'-TGACGTCA-3', homologous to sequences in other cAMP responsive genes. The two palindromes are contained within two identical 18-base pair (bp) sequences arranged as adjacent direct repeats. One or two synthetic copies of the 18-bp sequences were inserted into plasmids containing either the CG-alpha promoter or the SV40 promoter directing transcription of the chloramphenicol acetyltransferase gene. The 36-bp (double) element markedly enhanced chloramphenicol acetyltransferase activity in placental choriocarcinoma (JEG-3) cells when inserted in either orientation both 5' to the cap site or 3' of the coding sequence, thus defining it as an enhancer. Moreover, 8-br-cAMP stimulated the enhancer activity 30-40-fold. A single 18-bp element also stimulated chloramphenicol acetyltransferase activity, although 5-fold less than the double element, but still imparted a 35-fold transcriptional cAMP responsivity. The enhancer activates its homologous promoter much more efficiently than the SV40 promoter in JEG-3 cells. The alpha-promoter is not nearly as receptive to activation by the enhancer in baby hamster kidney fibroblasts, whereas the more modest enhancement of the SV40 promoter is less cell-specific. These studies suggest that the interaction of a 36-bp enhancer like element with the homologous promoter represents part of the mechanism of cell-specific expression of the CG-alpha gene and that the enhancer is co localized with a highly effective cAMP-response element. PMID- 3040733 TI - Nucleotide sequence of a member of the napin storage protein family from Brassica napus. AB - We have begun the molecular characterization of genes encoding napin, the 1.7 S embryo-specific storage protein of Brassica napus. Genomic Southern blot analysis indicates that napin is encoded by a multigene family comprised of a minimum of 16 genes. Two DNA fragments containing single napin genes have been recovered from B. napus genomic libraries. We have determined the nucleotide sequence of one member of the napin gene family, gNa. The gene has a simple structure lacking introns and containing the canonical features expected for genes transcribed by RNA polymerase II. The site of the initiation of transcription was determined to be 37 base pairs upstream of the initiation codon by S1 and primer extension analyses. A gene-specific hybridization probe from the 3' non-translated portion of gNa was used to demonstrate transcription of gNa. PMID- 3040734 TI - The hisT-purF region of the Escherichia coli K-12 chromosome. Identification of additional genes of the hisT and purF operons. AB - A 9.7-kilobase pair segment of the Escherichia coli chromosome spanning the hisT and purF loci has been characterized. Six genes were identified in this region by complete DNA sequence analysis, in vivo expression in maxicells, and RNA transcript analysis. S1 nuclease analysis has demonstrated that some of these genes are part of the hisT or purF operons. Two of the newly identified genes, dedA and dedB, were localized immediately downstream of hisT in the hisT operon. Two other genes, denoted dedC and dedD, have been localized between the hisT and purF operons. The other two genes, dedE and dedF flank the purF gene. dedE has been previously described as the first gene in the purF operon (Makaroff, C.A., and Zalkin, H. (1985) J. Biol. Chem. 260, 10378-10387). dedF was localized downstream from purF and is part of the purF operon. In addition, dedF is homologous to the ubiX gene of Salmonella typhimurium. Adjacent to dedF is the E. coli homologue of the S. typhimurium argT locus encoding the lysine/arginine/ornithine-binding protein. All of the genes in this region of the chromosome were found to be transcribed in a counter-clockwise direction on the E. coli map which revises the direction of purF transcription. PMID- 3040735 TI - Polyphosphoinositide micelles and polyphosphoinositide-containing vesicles dissociate endogenous gelsolin-actin complexes and promote actin assembly from the fast-growing end of actin filaments blocked by gelsolin. AB - The Ca2+-activated actin-binding protein gelsolin regulates actin filament length by severing preformed filaments and by binding actin monomers, stabilizing nuclei for their assembly into filaments. Gelsolin binds to phosphatidylinositol 4,5 bisphosphate (PIP2), with consequent inhibition of its filament severing activity and dissociation of EGTA-resistant complexes made with rabbit macrophage or human plasma gelsolin and rabbit muscle actin. This study provides evidence for an interaction of gelsolin with phosphatidylinositol monophosphate (PIP) as well as PIP2 and further describes their effects on gelsolin's function. Both phosphoinositides completely dissociate EGTA-insensitive rabbit macrophage cytoplasmic gelsolin-actin complexes and inhibit gelsolin's severing activity. The magnitude of inhibition depends strongly on the physical state of the phosphoinositides, being maximal in preparations that contain small micelles of either purified PIP or PIP2. Aggregation of PIP or PIP2 micelles by divalent cations or insufficient sonication or their incorporation into vesicles containing other phospholipids decreases but does not eliminate the inhibitory properties of the polyphosphoinositides. The presence of gelsolin partly inhibits the divalent cation-induced aggregation of PIP2 micelles. PIP2 in combination with EGTA inactivates gelsolin molecules that block the fast-growing end of actin filaments, thereby accelerating actin polymerization. Regulation of gelsolin by the intracellular messengers Ca2+ and polyphosphoinositides allows for the formation of several different gelsolin-actin intermediates with distinct functional properties that may be involved in changes in the state of cytoplasmic actin following cell stimulation. PMID- 3040736 TI - Mitochondrial and cytoplasmic fumarases in Saccharomyces cerevisiae are encoded by a single nuclear gene FUM1. AB - Respiratory defective pet mutants of Saccharomyces cerevisiae assigned to complementation group G5 are deficient in fumarase. A representative mutant from this complementation group was used to clone a nuclear gene (FUM1) whose sequence encodes a protein homologous to bacterial fumarase. Based on the primary structure homology and the elevated levels of fumarase in transformants harboring FUM1 on a multicopy plasmid, this gene is concluded to code for yeast fumarase. In wild type yeast, fumarase is detected in both mitochondria and the soluble postribosomal protein fraction. Several lines of evidence indicate that the two compartmentally distinct fumarases are isoenzyme products of FUM1. Mutations in FUM1 simultaneously abolish both activities. Transformation of a fumarase mutant with a plasmid containing FUM1 leads to increased fumarase activity in mitochondria and in the postribosomal supernatant fraction. Transformation of the same mutant with a plasmid construct in which the region of FUM1 coding for the amino-terminal 17 amino acids of fumarase is deleted results in a preferential increase of nonmitochondrial fumarase. Northern and S1 nuclease analysis of fumarase transcripts in wild type yeast and in a mutant transformed with FUM1 on an episomal plasmid indicate that the gene is transcribed from multiple start sites, some of which are located inside the coding sequence. The major transcript presumed to code for mitochondrial fumarase has a 5'-untranslated leader of 185 nucleotides. The most abundant shorter transcripts have 5' termini from 57 to 68 nucleotides downstream of the first ATG; their translation products lacking the amino-terminal mitochondrial import signal are proposed to target fumarase to the cytoplasm. PMID- 3040737 TI - Identification and characterization of the dihydropyridine-binding subunit of the skeletal muscle dihydropyridine receptor. AB - Photoaffinity labeling of isolated triads and purified dihydropyridine receptor with [3H]azidopine and (+)-[3H]PN200-110 has been used to identify and characterize the dihydropyridine-binding subunit of the 1,4-dihydropyridine receptor of rabbit skeletal muscle. The 1,4-dihydropyridine receptor purified from rabbit skeletal muscle triads contains four protein subunits of 175,000, 170,000, 52,000, and 32,000 Da (Leung, A., Imagawa, T., and Campbell, K. P. (1987) J. Biol. Chem. 262, 7943-7946). Photoaffinity labeling of isolated triads with [3H]azidopine resulted in specific and covalent incorporation of [3H]azidopine into only the 170,000-Da subunit of the dihydropyridine receptor and not into the 175,000-Da glycoprotein subunit of the receptor. The [3H]azidopine-labeled 170,000-Da subunit was separated from the 175,000-Da glycoprotein subunit by sequential elution from a wheat germ agglutinin-Sepharose column with 1% sodium dodecyl sulfate followed by 200 mM N-acetylglucosamine. Photoaffinity labeling of purified dihydropyridine receptor with [3H]azidopine or (+)-[3H]PN200-110 also resulted in the specific and covalent incorporation of either ligand into only the 170,000-Da subunit. Therefore, our results show that the dihydropyridine-binding subunit of the skeletal muscle 1,4-dihydropyridine receptor is the 170,000-Da subunit and not the 175,000-Da glycoprotein subunit. PMID- 3040738 TI - The function of superoxide dismutase during the enzymatic formation of the free radical of ribonucleotide reductase. AB - An enzyme system from Escherichia coli activates an inactive form of ribonucleotide reductase by transforming a tyrosine residue of the enzyme into a cationic free radical. The process requires NAD(P)H, a flavin, dithiothreitol, and oxygen and at least three proteins. After purification to near homogeneity two of the proteins were identified as superoxide dismutase and NAD(P)H:flavin oxidoreductase (Fontecave, M., Eliasson, R., and Reichard, P. (1987) J. Biol. Chem. 262, 12325-12331). The nature of the third protein, provisionally named Fraction b, is unknown. The flavin reductase is believed to reduce the ferric iron center of the ribonucleotide reductase as a prerequisite for radical generation. Here we demonstrate that the flavin reductase under aerobic conditions generates superoxide anions which inactivate ribonucleotide reductase. Superoxide dismutase protects the enzyme or a sensitive intermediate formed during the generation of the tyrosyl radical from the harmful effects of superoxide. Hydrogen peroxide, formed by superoxide dismutase, is also harmful. In this case, catalase present in Fraction b might protect the system. Fraction b has, however, an additional unknown function in the overall process of radical generation. PMID- 3040739 TI - Primary structure of the Escherichia coli folC gene and its folylpolyglutamate synthetase-dihydrofolate synthetase product and regulation of expression by an upstream gene. AB - The nucleotide sequence of the gene for folylpoly-gamma-glutamate synthetase dihydrofolate synthetase (folC) has been determined. The deduced amino acid sequence codes for a protein of Mr 45,380 and contains regions with homology to the A and B regions of ATP-binding sites. The folC gene is adjacent to a gene located 70 base pairs upstream of the initiation codon for folC. The nucleotide sequence of this upstream gene was also determined. Deletion of the upstream gene sequences from recombinant plasmids containing the folC gene results in about a 100-fold decrease in plasmid-dependent folylpolyglutamate synthetase activity, suggesting that the major promoter for folC is 5' to the upstream gene. The upstream gene codes for a protein of Mr 33,346, which is expressed in maxicells and amplified in cells containing the upstream gene in recombinant pUC8 plasmids. Expression of the upstream gene in maxicells was greater than that of folC, as determined by the intensity of 35S-labeled proteins after sodium dodecyl sulfate gel electrophoresis. A region of dyad symmetry exists between the coding sequences of the two genes which may code for a transcription termination signal and be responsible for the attenuation of the expression of the folC gene relative to the upstream gene. The folC gene is located about 1 kilobase upstream of the purF gene region at 50 min on the Escherichia coli map. The function of the upstream gene product is unknown. It contains sequences with homology to metal-binding domains in nucleic acid-binding proteins. A new purification procedure for obtaining large quantities of folylpolyglutamate synthetase dihydrofolate synthetase is described. PMID- 3040740 TI - Induction of proline-rich proteins in hamster salivary glands by isoproterenol treatment and an unusual growth inhibition by tannins. AB - Treatment of hamsters with the beta-agonist isoproterenol caused a dramatic increase in a series of unusual proteins in the parotid and submandibular glands. These proteins are acid soluble and they contain high amounts (mol%) of glutamate plus glutamine (30-35), proline (23-30), and glycine (12-25). Three proteins (HP45, HP43a, and HP43b) were isolated from trichloroacetic acid extracts of parotid glands of isoproterenol-treated hamsters. The basic protein (HP45) was not retained by DEAE-cellulose and did not contain phosphate or carbohydrate. Two acidic proteins (HP43a and HP43b) had the same apparent molecular weight on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, but these were separated by DEAE-cellulose chromatography. HP43a and HP43b contained 4.3 and 5.7 phosphate residues/mol of protein, respectively. Levels of mRNAs encoding this series of proteins showed striking increases following isoproterenol treatment as determined by cell-free translations and Northern analysis. Feeding tannins to rats and mice mimicks the effects of isoproterenol treatment on the parotid gland (Mehansho, H., Hagerman, A., Clements, S., Butler, L., Rogler, J., and Carlson, D.M. (1983) Proc. Natl. Acad. Sci. U.S.A. 80, 3948-3952; Mehansho, H., Clements, S., Sheares, B. T., Smith, S., and Carlson, D. M. (1985) J. Biol. Chem. 260, 4418 4423]. However, hamsters on a high tannin diet (2%) did not respond like rats and mice and instead displayed an unusual growth inhibition. Weanling hamsters maintained on a 2% tannin diet initially lost weight for 3 days and then failed to gain weight for up to 6 months when kept on this diet. Essentially a normal growth rate was observed when the tannin-fed hamsters were switched to a normal diet. PMID- 3040741 TI - Purification of the alpha 2-adrenergic receptor from porcine brain using a yohimbine-agarose affinity matrix. AB - A procedure has been developed for purification of the porcine brain alpha 2 adrenergic receptor to homogeneity. alpha 2-Adrenergic receptors were solubilized from porcine brain particulate preparations using sequential extraction into sodium cholate- and digitonin-containing buffers. The alpha 2-adrenergic receptors in the digitonin extract were identified using the alpha 2-adrenergic selective antagonist, [3H]yohimbine, and demonstrated the same specificity for interaction with adrenergic ligands as did the receptors in particulate preparations. Extraction into digitonin-containing buffers eliminated the modulation of receptor-agonist interactions by guanine nucleotides, but not by monovalent cations. A novel affinity resin, yohimbine-agarose, was synthesized and used for purification of alpha 2-adrenergic receptors. Using two sequential yohimbine-agarose affinity chromatography steps, digitonin-solubilized alpha 2 adrenergic receptors from porcine brain cortex were purified to homogeneity as assessed by radioiodination and silver stain analysis of these preparations on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The purified alpha 2 adrenergic receptor has an approximate Mr = 65,000, as determined by photolabeling of the adrenergic ligand-binding subunit. The yohimbine-agarose affinity resin should be useful for purifying quantities of receptor sufficient for studies of receptor structure and function. PMID- 3040743 TI - Structure-activity studies on synthetic peptides inhibiting herpes simplex virus ribonucleotide reductase. AB - Herpes simplex virus type 1 and type 2 (HSV-1 and HSV-2) ribonucleotide reductase is formed by the association of two nonidentical subunits. A peptide corresponding to the COOH terminus of the subunit 2, Tyr-Ala-Gly-Ala-Val-Val-Asn Asp-Leu (H2-(7-15)), has been shown to completely inhibit the reductase activity (IC50 = 36 microM) without affecting the host isoenzyme. In order to study the relationship between chemical requirements and inhibitory potencies, a series of peptides, including fragments and analogs of H2-(7-15), were synthesized. The minimum active core can be assigned to the Val-Val-Asn-Asp-Leu sequence (IC50 = 760 microM). N alpha-Extended peptides, such as Ser-Thr-Ser-Tyr-Ala-Gly-Ala-Val Val-Asn-Asp-Leu (H2-(4-15)) and Glu-Cys-Arg-Ser-Thr-Ser-Tyr-Ala-Gly-Ala-Val-Val Asn-Asp-Leu (H2-(1-15) ), respectively, have inhibitory potencies 2.1- and 1.4 fold greater than the nonapeptide H2-(7-15). N alpha-Deamination or acetylation of H2-(7-15) increases its potency by 1.8- and 3.0-fold, respectively, whereas amidation of the alpha-carboxylic function diminishes its activity by 3.2-fold. These results indicate that the alpha-amino group is not essential for maximum potency but suggest that a free carboxylic function is required. Substitution of Tyr7 or Ala8 by their respective D-isomer leads to a decrease of potency, suggesting that a specific conformation of the NH2-terminal portion is required to have a maximum activity. Monosubstitution in positions 11, 13, 14, and 15, by L-alanine completely abolishes activity stressing the importance of each amino acid residue contained in the minimum active core. Finally, nonapeptides corresponding to the COOH-terminal portion of the subunit 2 of Epstein-Barr and varicella-zoster virus ribonucleotide reductases also inhibit the HSV-1 reductase activity. The varicella-zoster virus nonapeptide is 4.0 times more potent than H2 (7-15), whereas the Epstein-Barr virus nonapeptide is 3.1 times less potent. These results should help us to design a new generation of potent inhibitors of herpes virus ribonucleotide reductases. PMID- 3040744 TI - Free radical intermediates in the reaction of pyruvate:ferredoxin oxidoreductase in Tritrichomonas foetus hydrogenosomes. AB - Aerobic incubations of the Tritrichomonas foetus hydrogenosomal fraction containing pyruvate, CoA, and the spin trap 5,5-dimethyl-1-pyrroline N-oxide (DMPO) gave spectra of two radical adducts. One was a carbon-centered radical adduct of DMPO. This radical was centered at C-3 of pyruvate as determined in experiments using [13C]pyruvate. The other radical detected was identified as the CoA radical adduct of DMPO by comparison with an adduct obtained by incubating CoA with DMPO, H2O2 and horseradish peroxidase. Deletion of CoA led to an increased stability of the carbon-centered radical adduct of DMPO, disappearance of the thiyl radical adduct of DMPO, and appearance of a hydroxyl radical adduct of DMPO. Superoxide dismutase suppressed the appearance of the DMPO-hydroxyl radical adduct but did not have any inhibitory effect on the appearance of the other adducts. Catalase had no significant effect on any of the adducts. Addition of pyruvate to these hydrogenosomal preparations stimulated oxygen consumption. Addition of CoA led to a further increase in the rate of O2 uptake but had no effect in the absence of pyruvate. The formation of two substrate free radicals as intermediates in the generation of acetyl-CoA represents a novel mechanism for this enzymatic reaction and indicates that the pyruvate:ferredoxin oxidoreductase from T. foetus differs significantly from the pyridine nucleotide-dependent pyruvate dehydrogenase complex of other eukaryotic cells in its catalytic mechanism. PMID- 3040742 TI - Regulation of porcine brain alpha 2-adrenergic receptors by Na+,H+ and inhibitors of Na+/H+ exchange. AB - Previous reports from this laboratory have demonstrated that alpha 2-adrenergic receptors accelerate Na+/H+ exchange in NG108-15 neuroblastoma X glioma cells and evoke platelet secretion via a pathway involving Na+/H+ exchange. The present studies were designed to examine whether agents that interact with Na+/H+ antiporters also might influence alpha 2-adrenergic receptor-ligand interactions. We observed that Na+ decreases receptor affinity for the agonists epinephrine, norepinephrine, and UK14304 and slightly increases receptor affinity for the antagonists yohimbine and idazoxan in digitonin-solubilized preparations from porcine brain cortex. Increases in [H+] also decrease receptor affinity for agonists and cause either a slight increase or no change in receptor affinity for antagonists. Amiloride analogs accelerate the rate of [3H] yohimbine dissociation from digitonin-solubilized receptors with a relative effectiveness that parallels their ability to block Na+/H+ exchange in other systems. Interestingly, these modulatory effects of Na+,H+ and 5-amino-substituted analogs of amiloride are retained in homogeneous preparations of the alpha 2-adrenergic receptor, suggesting that the allosteric-binding sites for these agents are on the receptor binding protein itself. PMID- 3040745 TI - Stereochemical requirements for substrate specificity of LTB4 20-hydroxylase. AB - LTB4 20-hydroxylase (P-450LTB) is the cytochrome P-450 in the microsomes of human polymorphonuclear leukocytes that catalyzes the omega-oxidation of leukotriene B4 (LTB4) to 20-OH LTB4. The activity of P-450LTB for LTB4 compared to isomers and analogs of LTB4 at a concentration of 0.3 microM revealed a preference of P 450LTB for both the triene bond configuration of LTB4 and for the chirality of the 5S and 12R hydroxyl groups. 15S-Hydroxyeicosatetraenoic acid, 8(R/S), 15S dihydroxy-5-cis-9,11,13-trans-eicosatetraenoic acid, 8R,15S-dihydroxy-5,13-cis 9,11-trans-eicosatetraenoic acid, and 5S,15S-dihydroxy-6,13-trans-8,11-cis eicosatetraenoic acid were each not subject to omega-oxidation, indicating a negative effect of the presence of a 15-hydroxyl group on substrate recognition. At a concentration of 1.5 microM, 12R- and 12S-hydroxyeicosatetraenoic acid were converted to their respective 20-OH derivatives at rates that were 34.2 +/- 11.6% (mean +/- S.D., n = 3) and 3.5 +/- 4.3% (mean +/- S.D., n = 4), respectively, of that of LTB4 to 20-OH LTB4, further indicating that P-450LTB can distinguish the chirality of the 12-hydroxyl group. The lower Km of LTB4 (2.0 microM), as compared to those of its 6-trans-12-epi isomer (3.8 microM) and 5-epi-LTB4 (6.6 microM) confirmed the preference of P-450LTB for the specific triene bond structure of LTB4 and its preference for the chirality of the hydroxyl groups of LTB4 within this structurally related class of molecules. At equal 1.5-microM concentrations, LTB4 completely inhibited the omega-oxidation of all other substrates and partially suppressed that of leukotriene B5, consistent with the lower Km of LTB4 and indicating that P-450LTB catalyzed the omega-oxidation of all substrates. Thus, P-450LTB is a novel cytochrome P-450 of human polymorphonuclear leukocytes with substrate recognition determined by the triene bond configuration and the chirality of the hydroxyl groups. PMID- 3040746 TI - Prosthetic group content and ligand-binding properties of spinach nitrite reductase. AB - Chemical analysis of the ferredoxin-dependent native form (Mr = 85,000) of spinach nitrite reductase has demonstrated a siroheme content that approaches 2 mol of siroheme/mol of enzyme. A widely studied modified (Mr = 61,000) form of nitrite reductase, that has lost much of the native enzyme's ability to use ferredoxin as an electron donor, contains approximately 1 mol of siroheme/mol of enzyme. Quantitation of the high spin ferri-siroheme EPR signals and of nitrite binding sites of the two preparations confirmed that the native enzyme's siroheme content is approximately twice that of the modified enzyme. Plots of nitrite and cyanide binding to the native enzyme versus ligand concentration are sigmoidal, with Hill coefficients of 1.6-1.8 and 2.3-2.8, respectively. Plots of enzyme activity versus nitrite concentration for the native enzyme are sigmoidal with a Hill coefficient of 2.4. Cyanide inhibition of enzymatic activity was shown to be not competitive. Addition of cyanide to the native enzyme resulted in a diminution of the high spin ferri-siroheme EPR signal and produced EPR signals with g values of 2.71, 2.33, and 1.49 due to low spin ferri-siroheme. PMID- 3040748 TI - Spin trap determination of free radical burst kinetics in stimulated neutrophils. AB - Electron spin resonance spin-trapping methods were used to investigate the free radical production kinetics of neutrophils stimulated with phorbol myristate acetate (PMA) and opsonized zymosan (OPZ). Using the spin trap 5,5-dimethyl-1 pyrroline-N-oxide, the principle spin adduct observed is DMPO-OH (trapped hydroxyl radical). The DMPO-OH ESR signal amplitude was observed to decay exponentially. In such cases a simple method may be used to analyze the raw kinetics amplitude data to yield true production rate and net production data. The method, pitfalls, and self-consistency criteria are illustrated with PMA and OPZ-stimulated neutrophils at 25 and 37 degrees C under varying oxygen tensions, and with noise-free simulated data. The simulations demonstrate that rate results are relatively insensitive to the precise choice of decay time constant, tc, while net production results are very sensitive to the choice of tc used to analyze the raw data. OPZ (0.6-2.4 mg/ml) yields a strong, sharp neutrophil burst which peaks in 2 min or less while PMA yields a slower burst which peaks in 3.4 14 min for PMA concentrations of 500-50 ng/ml, respectively. Increased oxygen tension during the PMA experiments increased the spin adduct lifetime. The methods presented are applicable to other cell systems or spin adducts which exhibit first order decay. PMID- 3040747 TI - Mechanism of energy coupling to entry and exit of neutral and branched chain amino acids in membrane vesicles of Streptococcus cremoris. AB - The energetics of neutral and branched chain amino acid transport by membrane vesicles from Streptococcus cremoris have been studied with a novel model system in which beef heart mitochondrial cytochrome c oxidase functions as a proton motive force (delta p) generating system. In the presence of reduced cytochrome c, a large delta p was generated with a maximum value at pH 6.0. Apparent H+/amino acid stoichiometries (napp) have been determined at external pH values between 5.5 and 8.0 from the steady state levels of accumulation and the delta p. For L-leucine napp (0.8) was nearly independent of the pH. For L-alanine and L serine napp decreased from 0.9-1.0 at pH 5.5 to 0-0.2 at pH 8.0. The napp for the different amino acids decreased with increasing external amino acid concentration. At pH 6.0, first order rate constants for amino acid exit (kex) under steady state conditions for L-leucine, L-alanine, and L-serine were 1.1 1.3, 0.084, and 0.053 min-1, respectively. From the pH dependence of kex it is concluded that amino acid exit in steady state is the sum of two processes, pH dependent carrier-mediated amino acid exit and pH-independent passive diffusion (external leak). The first order rate constant for passive diffusion increased with increasing hydrophobicity of the side chain of the amino acids. As a result of these processes the kinetic steady state attained is less than the amino acid accumulation ratio predicted by thermodynamic equilibrium. The napp determined from the steady state accumulation represents, therefore, a lower limit. It is concluded that the mechanistic stoichiometry (n) for L-leucine, L-alanine, and L serine transport most likely equals 1. PMID- 3040749 TI - Differential inactivation of inotropic and toxic digitalis receptors in ischemic dog heart. Molecular basis of the deleterious effects of digitalis. AB - When applied to ischemic hearts digitalis exhibits depressed inotropic effect and increased toxicity. The molecular basis of these effects was investigated at the level of the digitalis receptors characterized by Na,K-ATPase assays and [3H]ouabain-binding measurements. In sarcolemma obtained from dog hearts rendered ischemic for 15, 30, and 60 min (left anterior descending), two populations (high and low affinity) of digitalis receptors were detected. The apparent affinity (KD, 300 nM) and the binding capacity of the low-affinity sites (responsible for toxicity) remained constant and similar to those found in normal hearts. The KD value of the high-affinity sites, "responsible for inotropy," remained unchanged (2 nM), but the site number sharply decreased (up to 90%). These inotropic sites that account for 66% of the total binding in normals are gradually inactivated, as the duration of ischemia increases. This inactivation would occur in situ since it was detectable in homogenates and was not depressed by the isolation procedure per se. The loss of function of the inotropic sites and the increased contribution of the low-affinity toxic sites represent the setting of a new distribution of the digitalis receptors in the ischemic heart before reperfusion is instituted. This constitutes the molecular basis of the deleterious pharmacological effects observed with digitalis. PMID- 3040750 TI - Possible involvement of a GTP-binding protein, the substrate of islet-activating protein, in receptor-mediated signaling responsible for cell proliferation. AB - Serum-induced DNA synthesis, as measured by increases in [3H]thymidine incorporation, in Swiss mouse 3T3 fibroblasts was markedly inhibited by exposure of the cells to islet-activating protein (IAP), pertussis toxin. The inhibition was well correlated with the toxin-induced ADP-ribosylation of a membrane GTP binding protein with Mr = 41,000. The IAP-induced inhibition of cell growth was characterized by the following two features. First, the inhibition was selective to certain growth factors. DNA synthesis in 3T3 cells was supported by a combination of one of the competence factors and a progression factor such as insulin or epidermal growth factor. IAP was inhibitory when thrombin, fibroblast growth factor, prostaglandin F2 alpha, or phosphatidic acid was employed as a competence factor, but was not inhibitory when DNA synthesis was induced by combined addition of cholera toxin or phorbol ester with insulin. Second, IAP induced inhibition was still observed when the toxin was added to cell culture 1 6 h later than the addition of the IAP-sensitive competence factors, which triggered rapid cellular responses such as adenylate cyclase inhibition, releases of inositol trisphosphate and arachidonic acid, and 45Ca influx within several minutes (Murayama, T., and Ui, M. (1985) J. Biol. Chem. 260, 7226-7233; Murayama, T., and Ui, M. (1987) J. Biol. Chem. 262, 5522-5529). Thus, IAP substrate GTP binding protein(s) appears to be involved in the duration of rapid signals or the occurrence of new slow signals which are responsible for growth factor-induced cell proliferation. The site of the involvement may be proximal to protein phosphorylation by phorbol ester-activated and cAMP-dependent kinases. PMID- 3040751 TI - Purification and properties of an extracellular collagenolytic protease produced by the human oral bacterium Bacillus cereus (strain Soc 67). AB - The major collagenolytic proteinase present in the culture filtrate of Bacillus cereus (strain Soc 67, isolated from the human oral cavity) has been purified to homogeneity by a procedure that comprised concentration of ultrafiltered growth medium on a Millipore PTTK00005 membrane, precipitation with ammonium sulfate, gel permeation chromatography, chromatofocusing, fast protein liquid chromatography on an anion-exchange column, and finally fast protein liquid chromatography on a gel column. The enzyme hydrolyzed, with decreasing rates, phenylazobenzyloxy-carbonyl-L-Pro-L-Leu Gly-L-Pro-D-Arg (PZ-PLGPA), furylacrylolyl-L-Leu-Gly-L-Pro-L-Ala, and furylacryloyl-L-Phe-Gly-Gly, while furylacryloyl-Gly-L-Leu-NH2 was not hydrolyzed. The enzyme degraded soluble and insoluble collagens, Azocoll and gelatin. Bradykinin was hydrolyzed at a high rate at the Phe-Ser bond. The enzyme was sensitive to pyrophosphate, L-cysteine, and L-histidine and could be totally inactivated in the presence of metal chelators. The enzyme contains 1 mol of Zn/mol and the hydrolysis of PZ-PLGPA is slightly increased by Ca2+. The enzyme is readily inhibited by heavy metal cations, but Cu2+ and Ni2+ affected the catalysis in opposite ways: increasing levels of Cu2+ decreased the affinity of the enzyme for PZ-PLGPA, whereas Ni2+ had no effect. The effect of Cu2+ also depended on the pH and type of buffer used. Detailed chemical modification experiments suggested that the active site of the enzyme contains at least 1 tyrosyl and 1 lysyl residue, and 1 carboxyl group. The enzyme was not sensitive to sulfhydryl reagents and thiols did not activate the enzyme. The modification studies were unable to reveal active histidyl residues. The ability of the enzyme to hydrolyze PZ-PLGPA, furylacryloyl L-Leu-Gly-L-Pro-L-Ala, furylacryloyl-L-Phe-Gly-Gly, and various collagenous materials, its inactivity toward furylacryloyl-Gly-L-Leu-NH2, and the results from the chemical modification studies suggest that the B. cereus (Soc 67) collagenolytic enzyme can be regarded as a true collagenase which resembles the Clostridium histolyticum collagenase(s). PMID- 3040752 TI - Transport of the vesicular stomatitis glycoprotein to trans Golgi membranes in a cell-free system. AB - Terminal steps in the transport of the vesicular stomatitis virus glycoprotein (G protein) in the Golgi stack have been reconstituted in a cell-free system. Incorporation of sialic acid into the oligosaccharide chains of G protein was used to monitor transport into the trans Golgi compartment. Transport-coupled sialylation required cytosol, ATP, an N-ethylmaleimide-sensitive factor extractable from Golgi membranes, and long chain acyl coenzyme A. The G protein receiving sialic acid in the cell-free system begins its in vitro transport bearing galactose residues acquired in vivo. Earlier reports (Balch, W. E., Dunphy, W. G., Braell, W. A., and Rothman, J. E. (1984a) Cell 39, 405-416) documented that transport of G protein into the medial (GlcNAc Transferase containing) compartment is reconstituted under the same conditions. On the basis of the results reported here, it now appears that a more complete set of transport operations of the Golgi stack may be simultaneously reconstituted. PMID- 3040753 TI - Purification and characterization of two immunologically distinct phosphoinositide-specific phospholipases C from bovine brain. AB - We previously reported (Ryu, S. H., Cho, K. S., Lee, K. Y., Suh, P. G., and Rhee, S. G. (1986) Biochem. Biophys. Res. Commun. 141, 137-144) that cytosolic fractions of bovine brain contain two phosphoinositide-specific phospholipase C (PLC), PLC-I and PLC-II. In this paper purification procedures and properties of these two forms of enzyme are presented. The two enzymes exhibit similar substrate specificity. Both PLC-I and PLC-II catalyze the hydrolysis of phosphatidylinositol (PI), phosphatidylinositol-4-phosphate (PIP), and phosphatidylinositol-4,5-bisphosphate (PIP2). Yet, they respond differently to activators such as Ca2+ and nucleotides and to inhibitory divalent metal ions such as Hg2+ and Cd2+. In addition, they are immunologically distinct as evidenced by the fact that monoclonal antibodies directed against either enzyme do not cross-react with the other. Their activities are Ca2+ concentration dependent. PIP and PIP2 are better substrates than PI for both PLC-I and PLC-II when the concentration of Ca2+ is in the micromolar range. Study of the effect of nucleotides, such as GTP, guanosine 5'-(3-O-thio)triphosphate, guanyl-5'-yl imidodiphosphate, and ATP, on the activities of both isozymes with PIP2 as substrate revealed that (i) in the absence of Ca2+, PLC-I activity is enhanced by 400% by either GTP or ATP. In the presence of Ca2+ (a condition in which PLC-I exhibits much higher activity), the activation factor by nucleotides is diminished to approximately 140%. (ii) without Ca2+, PLC-II activity is too low to measure with or without added nucleotides. The effect of nucleotides on PLC-II activity is trivial in the presence of Ca2+. In addition, studies on the effect of metal ions on PI hydrolysis showed that the activities of both PLC-I and PLC II are not affected by 50 microM of Mg2+, Mn2+, Ca2+, or Ni2+. However, Hg2+, Zn2+, and Cu2+ inhibited both PLC-I and PLC-II, with PLC-II exhibiting much higher sensitivity to these metal ions than PLC-I. For example, the value of I0.5 for Hg2+ inhibition is 0.2 microM for PLC-II and 1 microM for PLC-I. Cd2+ selectively inhibits PLC-II with a I0.5 value of 5 microM. Most of these metal ions' inhibition can be overcome by either dithiothreitol or EDTA. PMID- 3040754 TI - Purification of a phosphoinositide-specific phospholipase C from bovine brain. AB - A soluble phosphoinositide-specific phospholipase C (PLC) was purified 58,000 fold from bovine brain. The enzyme, one of six distinct PLC activities detected in brain, accounted for approximately 15% of the soluble phosphatidylinositol-4,5 bisphosphate-phospholipase C (PIP2-PLC) activity in this tissue. The purification scheme included hydrophobic chromatography on phenyl-Sepharose and affinity chromatography on phosphatidylinositol-Sepharose (PI-Sepharose). The enzyme was specifically eluted from the PI-Sepharose with PI, calcium, and detergent. The purified PLC had an estimated molecular weight of 88,000 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and behaved as a monomeric protein during sedimentation on glycerol gradients. The enzyme required calcium for activity, exhibited a pH optimum of 6.5, and cleaved only phosphoinositides. The rates of PIP2 and phosphatidyl-4-monophosphate hydrolysis exceeded the rate of PI hydrolysis under all conditions tested. These properties are consistent with a potential role for this PLC in the early events involved in cellular calcium mobilization. PMID- 3040755 TI - Effects of gentamicin on sphingomyelinase activity in cultured human renal proximal tubular cells. AB - We have previously shown that cultured human proximal tubular cells (PT) incubated with gentamicin contain numerous "myeloid bodies." This morphological change was accompanied by the storage of phosphatidylcholine and sphingomyelin. In order to delineate the biochemical mechanisms responsible for the accumulation of sphingomyelin in cells incubated with gentamicin, we pursued detailed studies on the activity of sphingomyelinase. Characterization studies on sphingomyelinase revealed that this enzyme has a bimodal pH optima in PT cells. Optimum activity was observed at pH 5.6 (designated as acid sphingomyelinase, A-SMase) and at pH 7.4 (designated as neutral sphingomyelinase, N-SMase). The activity of both the enzymes increased proportionately in control cells as a function of days of incubation. The activity of A-SMase was 16% lower in cells incubated with gentamicin as compared to control. The most striking observation was a gradual decline in the activity of N-SMase in cells incubated with gentamicin. Thus, following 21 days of incubation of cells with 0.3 mM gentamicin, the N-SMase was 2.7-fold lower than control cells. Mg2+ stimulated and Triton X-100 inhibited the activity of N-SMase. Whereas Mg2+ had no effects, Triton X-100 stimulated the activity of the A-SMase in PT cells. Moreover, A-SMase was relatively more heat resistant than the N-SMase. The Km values for sphingomyelin using A-SMase in control cells and cells incubated with gentamicin were 0.07 X and 0.016 X 10(-7) M, respectively, whereas the Km values for sphingomyelin using N-SMase in control cells and cells incubated with gentamicin were 1.8 X and 1.5 X 10(-7) M, respectively. These findings suggest that gentamicin exerts a competitive inhibition of the A-SMase in PT cells. In contrast, gentamicin exerts a noncompetitive inhibition of the N-SMase in PT cells. Subcellular fractionation studies revealed that A-SMase was exclusively localized in the "lysosome-rich" fraction, whereas most, if not all, the N-SMase was localized in the microsomal fraction and "plasma-membrane"-rich fraction in cultured PT cells. Cells incubated with gentamicin for 21 days contained 25% lower activity of A-SMase associated with the lysosomal fraction as compared to control. In contrast, N SMase activity in the microsomal and plasma membrane fraction was one-half as compared to control. We conclude that gentamicin-mediated decrease in sphingomyelinase activity may be responsible for the storage of sphingomyelin in cultured human PT cells.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3040756 TI - Cholesterol flux between cells and high density lipoprotein. Lack of relationship to specific binding of the lipoprotein to the cell surface. AB - The bidirectional flux of unesterified cholesterol between cells and high density lipoprotein (HDL) was studied in relationship to the binding of HDL to cells. At 100 micrograms at HDL protein/ml, the rate constant for cholesterol efflux from rat Fu5AH hepatoma cells is 3 X 10(-3)/min (t1/2 for efflux of 3.9 h), whereas efflux from GM3468 human fibroblasts is 0.075/4 h (equivalent to a t1/2 for efflux of 37 h). The relatively slow efflux of cholesterol from fibroblasts in comparison to rat hepatoma cells was observed previously with micellar and vesicular phospholipid-containing acceptors, which promote efflux by a mechanism involving the diffusion of cholesterol in the aqueous phase between the plasma membrane and the acceptor particles. When plotted against the logarithm of HDL concentration, the isotherms for efflux are centered at 300 and 100 micrograms of HDL protein/ml with the hepatoma cells and fibroblasts, respectively. These concentrations are 8-150 times greater than the corresponding values for Kd of specific HDL binding (2 and 12 micrograms of protein/ml, for hepatoma cells and fibroblasts, respectively). The treatment of HDL with tetranitromethane reduces the lipoprotein's affinity for specific cell-surface binding sites by 80-90%. However, at HDL concentrations of 5-60 micrograms of protein/ml, this treatment does not significantly inhibit cholesterol efflux from hepatoma cells, and inhibits efflux from fibroblasts an average of about 15%. Over the same range of concentrations, nitration alters influx by amounts less than 30% in the two cell types. These effects on flux do not parallel the reduced affinity of nitrated HDL for specific cell-surface binding sites. In summary, the present results do not support the concept that cholesterol transfer is facilitated by the specific cell surface binding of HDL, but are consistent with the aqueous diffusion model of cholesterol transfer between cells and lipoproteins. PMID- 3040757 TI - Evidence for inherent differences in the system A carrier from normal and transformed liver tissue. Differential inactivation and substrate protection in membrane vesicles and reconstituted proteoliposomes. AB - Plasma membrane vesicles isolated from intact rat liver (normal hepatocyte) or cultured rat H4 hepatoma cells retain Na+-dependent uptake of 2-aminoisobutyric acid mediated by System A. The carrier was inactivated in normal liver membrane vesicles by either N-ethylmaleimide (NEM) or p-chloromercuribenzene sulfonate (PCMBS). The concentrations required to produce half-maximal inhibition were approximately 370 and 110 microM for NEM and PCMBS, respectively. In contrast, transport of System A in H4 hepatoma membrane vesicles was sensitive to PCMBS (K 1/2 = 180 microM), yet totally unaffected by NEM at concentrations up to 5 mM. Substrate-dependent protection from PCMBS activation was observed for the System A activity in H4 hepatoma membranes, but not in vesicles from normal hepatocytes. Subsequent inactivation of the substrate-protected carrier by sulfhydryl-specific reagents, added following the removal of the protective amino acid, suggests that one or more cysteine residues become less reactive in the presence of System A substrates. Treatment of solubilized membrane proteins with NEM prior to reconstitution into artificial proteoliposomes showed that the selective inactivation by NEM of the carrier in normal liver membranes is not dependent on the lipid environment or on the integrity of the plasma membrane. The results support the hypothesis that there are inherent differences in the System A carriers that are present in normal and transformed liver tissue. PMID- 3040758 TI - Chemical modification of the bifunctional regulatory protein of maize leaf pyruvate,orthophosphate dikinase. Evidence for two distinct active sites. AB - The active site(s) of the bifunctional regulatory protein of pyruvate,orthophosphate dikinase catalyze(s) the Pi-dependent activation (dephosphorylation) and ADP-dependent inactivation (phosphorylation) of maize leaf dikinase. The chemical modification studies of the regulatory protein active sites presented in this paper are interpreted as showing the two sites to be physically distinct. Pyridoxal 5'-phosphate and 2-nitro-5-thiocyanatobenzoate (NTCB) selectively inhibit the dikinase activating site, which is protected by the nonprotein substrate, Pi. Phenylglyoxal blocks both the activation and inactivation sites; the former is protected selectively by Pi and the latter by both the nonprotein substrate, ADP, and Pi. The Pi that protects the inactivation site is distinct from the activation substrate. Inhibition studies show Pi to be a parabolic competitive inhibitor of the ADP-dependent inactivation of dikinase, implying that besides substrate Pi, a second phosphate also binds to the regulatory protein. The above chemical modifications are not mutually exclusive; neither NTCB, 5,5'-dithiobis-(2-nitrobenzoate), nor pyridoxal 5'-phosphate blocks subsequent modification of the activation site by phenylglyoxal. Similarly, prior modification with NTCB does not affect modification by pyridoxal 5'-phosphate. PMID- 3040759 TI - Priming of neutrophils and macrophages for enhanced release of superoxide anion by the calcium ionophore ionomycin. Implications for regulation of the respiratory burst. AB - Phagocytic cells can be primed for enhanced stimulated release of superoxide anion (O2-) by exposure to a variety of biologic agents, including gamma interferon and lipopolysaccharide. We examined the role of calcium ion in this priming, using the calcium ionophore ionomycin. Preincubation with ionomycin, 1 to 10 nM, primed human neutrophils to release up to 7-fold more O2- during stimulation with 1 microM formyl-methionyl-leucyl-phenylalanine (f-Met-Leu-Phe). With 160 nM phorbol myristate acetate as stimulus, ionomycin caused a doubling of O2- production in mouse peritoneal macrophages. Incubation of phagocytes with ionomycin at priming concentrations did not directly stimulate O2- release. Priming of neutrophils occurred in 1-2 min and was associated with a marked reduction in the lag time for O2- release after f-Met-Leu-Phe stimulation and with an increase in the rate of O2- production. Kinetic analysis of NADPH dependent O2(-)-producing activity in sonicates of resting human neutrophils incubated with sodium dodecyl sulfate suggested that modification of the enzyme responsible for the respiratory burst was not responsible for priming. Priming of neutrophils with ionomycin had no apparent effect on either the activity or subcellular distribution of protein kinase C. The effect of ionomycin on the cytosolic free calcium concentration ([Ca2+]c) was assessed in neutrophils using the calcium-sensitive fluorescent dye fura-2. Ionomycin at priming concentrations caused an approximate doubling of the base-line [Ca2+]c. When neutrophils were exposed to various concentrations of ionomycin, a parallel rise in [Ca2+]c and priming was observed. A rise in [Ca2+]c of approximately 0.8 microM caused half maximal priming. These results suggest that an increase in [Ca2+]c is not sufficient to initiate release of O2-, but they support the concept that Ca2+ can serve as a second messenger in this event. PMID- 3040760 TI - Autoxidation of oxymyoglobin. An overall stoichiometry including subsequent side reactions. AB - Oxymyoglobin (MbO2) is oxidized easily to metmyoglobin (metMb) with generation of the superoxide anion, which can be converted by the spontaneous dismutation into H2O2, this being also a potent oxidant of MbO2. In the presence of sodium azide in stoichiometric amounts, however, the rate of autoxidation of MbO2 increased rapidly with increasing concentration of the anion, but soon reached a saturating level, the extent of which was about twice that of the normal autoxidation in buffer alone. Quantitative analysis has revealed that this enhancement is not due to the nucleophilic displacement of O2- from MbO2 by the anion (Satoh, Y., and Shikama, K. (1981) J. Biol. Chem. 256, 10272-10275), but is due to the additional oxidation of MbO2 by H2O2 freed from the metMb being occupied by the anion at the sixth coordination position. Based on these novel results and stoichiometric considerations, it is possible to propose a new view that H2O2 produced from O2- can be eliminated or decomposed mostly, if not completely, by the metMb resulting from the normal autoxidation reaction of MbO2, presumably via the formation of the ferryl species. PMID- 3040761 TI - Identification of glycoprotein Ib beta as one of the major proteins phosphorylated during exposure of intact platelets to agents that activate cyclic AMP-dependent protein kinase. AB - Platelet function is inhibited by prostaglandin E1, prostaglandin I2, or forskolin, agents that increase the intracellular concentration of cyclic AMP. The inhibition appears to result from cyclic AMP-stimulated phosphorylation of specific intracellular proteins. One of the major increases in phosphorylation occurs in a polypeptide of Mr = 24,000 (P24). In this study, an effort was made to identify P24. Platelets prelabeled with [32P]phosphate were incubated with prostaglandin E1, prostaglandin I2, or forskolin. Proteins that became phosphorylated were detected by autoradiography of sodium dodecyl sulfate polyacrylamide gels. Several lines of evidence indicated that P24 was the beta subunit of the plasma membrane glycoprotein (GP) Ib, a glycoprotein that is essential for the adhesion of platelets to damaged subendothelium, for the rapid response of platelets to thrombin, and for the attachment of the membrane skeleton to the cytoplasmic face of the plasma membrane. P24 co-migrated with GP Ib beta on reduced gels (Mr = 24,000) and also on nonreduced gels (when GP Ib beta is disulfide-linked to GP Ib alpha and migrates with Mr = 170,000). Like GP Ib beta, P24 was associated with actin filaments in Triton X-100 lysates. Like GP Ib beta, it was selectively associated with filaments of the membrane skeleton and was released from filaments when the Ca2+-dependent protease was active. Antibodies against GP Ib immunoprecipitated P24 from platelet lysates. Finally, exposure of Bernard-Soulier platelets (which lack GP Ib) to prostaglandin E1 resulted in phosphorylation of other polypeptides, but not of P24. These studies show that P24, one of the major polypeptides phosphorylated when platelets are exposed to agents that inhibit platelet function by increasing the concentration of cyclic AMP, is the beta-subunit of GP Ib. PMID- 3040762 TI - Active site sequence of hepatic fructose-2,6-bisphosphatase. Homology in primary structure with phosphoglycerate mutase. AB - The reaction mechanism of rat hepatic fructose-2,6-bisphosphatase involves the formation of a phosphohistidine intermediate. In order to determine the sequence around the active site histidine, the enzyme was incubated with [2-32P]fructose 2,6-bisphosphate, denatured, and treated with trypsin or endoproteinase Lys-C. The resultant labeled 32P-phosphopeptides were purified by gel filtration, anion exchange chromatography, and reverse phase high pressure liquid chromatography. The sequence of the tryptic peptide was determined to be HGESELNLR, while the partial sequence of the endoproteinase Lys-C peptide was IFDVGTRYMVNRVQDHVQSRTAYYLMNIHVTPRSIYLRHGESEL. The active site sequence was compared with the active site sequence of other enzymes that catalyze phospho group transfer via a phosphohistidine intermediate. Active site sequences of phosphoglycerate mutase and bisphosphoglycerate synthase were highly homologous with the active site of fructose-2,6-bisphosphatase implying a structural similarity and a common evolutionary origin. PMID- 3040763 TI - Monoclonal antibodies specific for the tau subunit of the DNA polymerase III holoenzyme of Escherichia coli. Use to demonstrate that tau is the product of the dnaZX gene and that both it and gamma, the dnaZ gene product, are integral components of the same enzyme assembly. AB - We have established two murine hybridoma cell lines that secrete monoclonal antibodies directed against the tau subunit of the DNA polymerase III holoenzyme of Escherichia coli. Both antibodies have been purified and identified to be of the IgG1 class. Competition assays indicate that they bind to two distinct portions of the tau subunit. These antibodies have been used to demonstrate that tau is an integral part of all DNA polymerase III holoenzyme assemblies and that tau is the product of the dnaZX gene. Both of the antibodies react only with tau, not with gamma, the other protein product of the dnaZX gene. Immunoprecipitation studies demonstrated that tau is contained within the same enzyme assemblies as gamma (dnaZ protein). This observation is discussed in the light of the DNA polymerase III holoenzyme functioning as an asymmetric dimer, capable of coordinating leading with lagging strand replication. PMID- 3040764 TI - Possible involvement of lumichrome in the binding of storage protein to its receptor in Sarcophaga peregrina. AB - Previously, a receptor molecule on the surface of fat body cells of Sarcophaga peregrina larvae that is involved in the uptake of storage protein from the hemolymph was shown to be a 120-kDa protein (Ueno, K., and Natori, S. (1984) J. Biol. Chem. 259, 12107-12111). This paper reports evidence that lumichrome (7,8 dimethylalloxazine) may be present in the binding site of the receptor and mediate the binding of storage protein and receptor. A stoichiometric amount of lumichrome was shown to bind to the storage protein under conditions in which the latter bound to its receptor. PMID- 3040765 TI - Decrease in glucokinase and glucose-6-phosphatase and increase in hexokinase in putative preneoplastic lesions of rat liver. AB - Preneoplastic liver lesions were produced in female Wistar rats by oral administration of 2-acetylaminofluorene for 165 days succeeded by a carcinogen free standard diet up to 420 days. During the treatment numerous altered hepatic foci (AHF) and hyperplastic nodules (HN) were detected histochemically by a focal decrease or lack of adenosine-5-triphosphatase and glucose-6-phosphatase (G-6 Pase) activities. In addition, the immunohistochemically demonstrable amount of L type pyruvate kinase was clearly reduced. The histochemically demonstrated decrease of G-6-Pase was substantiated by microbiochemical determination of the enzyme activity in microdissected material. Moreover, during the experimental period a continuous decrease in glucokinase and an increase in hexokinase was detected microbiochemically within AHF and HN. These alterations indicate a shift in the carbohydrate metabolism from gluconeogenesis to glucose utilization and pentose-phosphate-pathway for biosynthesis of nucleic acids. Beside other oncofetal markers, HK may be used as indicator of the early stages of liver carcinogenesis. PMID- 3040766 TI - Hepatocellular carcinoma and hepatitis B virus infection: molecular evidence for monoclonal origin and expansion of malignantly transformed hepatocytes. AB - The clonality of tumor cells was studied in a patient with metastasizing hepatocellular carcinoma (HCC). Using hepatitis B virus (HBV) DNA as a genetic marker, the pattern of integration of viral DNA into the tumor cell genome was determined by Southern blot analyses of DNAs extracted from different HCC lesions in the liver and both lungs. All tumor tissues examined were found to have viral DNA integrated into the same site(s) of the cellular genome. This finding provides direct molecular evidence for a monoclonal origin and expansion of malignantly transformed hepatocytes during tumor growth and metastasis. This characteristic is similar to other human cancers associated with viral infections, such as adult T-cell leukemia, Burkitt's lymphoma, or cervical cancer, and is important for our understanding of viral oncogenesis in man. PMID- 3040767 TI - TNM stage, immunohistology, syntactic structure analysis and survival in patients with small cell anaplastic carcinoma of the lung. AB - TNM stage, immunostaining with various monoclonal and polyclonal antibodies, analysis of distance of neighboring cells, remission rates, and survival were analyzed in 60 patients suffering from small cell anaplastic carcinoma of the lung. The majority of patients showed advanced tumors at the time of admission to hospital (T2, T3 stage). Distant metastases prior to chemotherapy were detected in 34 patients. Partial remissions lasting 2-4 months were observed in 38 patients, and complete remission was documented in 7 patients. The remission rate was independent of cell type but dependent on the stage of the tumor. Some 30 patients showed positive staining with an antibody recognizing epitopes detectable on carcinoembryonic antigen, whereas 60% of the tumors were positive to a polyclonal neuron-specific enolase antibody. Tissue polypeptide antigen was found to stain positively in 5 cases only. Some 14 patients with negative staining against the monoclonal antibody BMA 406/14 showed prolonged survival compared to patients with positive staining (P less than 0.05). Patients suffering from tumors with smaller distances between neighboring cells had worse prognoses compared to patients with larger distances (P less than 0.01). Survival of patients was found to be indistinguishable if cohorts were grouped according to T stage, N stage, or existence of distant metastases. Ten patients who underwent surgical treatment of tumors did not show prolonged survival compared to 50 patients treated by combined chemotherapy only. PMID- 3040768 TI - Nipple involvement and multicentricity in breast cancer. A study on whole organ sections. AB - Our study examined 166 patients with breast cancer with a mean age of 63 years. Each patient underwent mastectomy with the organ being investigated by histological giant sections and additional small sections from the nipple. Nipple involvement was found in 64 cases (38%). Multifocal carcinoma occurred in 76 patients. Further multicentric carcinomatous foci (36 cases) demonstrated a significant increase in affected nipples. Additional atypical ductal or lobular hyperplasia was observed in 53 cases and showed involvement in 34. Nine carcinomas of ductal origin were combined with lobular carcinoma in situ, all cases proved to have carcinomatous changes in the nipple. It is concluded that apart from the well-known influence of advanced tumor stages and tumor localization, nipple involvement correlates with multicentricity and multifocality of breast cancer as a disease of the whole organ. PMID- 3040769 TI - Identification and characterization of a mouse cell mutant defective in the intracellular transport of glycoproteins. AB - We have isolated a mutant line of mouse L cells, termed gro29, in which the growth of herpes simplex virus (HSV) and vesicular stomatitis virus (VSV) is defective. The block occurs late in the infectious cycle of both viruses. We demonstrate that HSV and VSV enter gro29 cells normally, negotiate the early stages of infection, yet are impaired at a late stage of virus maturation. During VSV infection of the mutant cell line, intracellular transport of its glycoprotein (G protein) is slowed. Pulse-chase experiments showed that oligosaccharide processing is impeded, and immunofluorescence localization revealed an accumulation of G protein in a juxtanuclear region that contains the Golgi complex. We conclude that export of newly made glycoproteins is defective in gro29 cells, and speculate that this defect may reflect a lesion in the glycoprotein transport apparatus. PMID- 3040770 TI - In vitro mutagenesis of trypsinogen: role of the amino terminus in intracellular protein targeting to secretory granules. AB - The mouse anterior pituitary tumor cell line, AtT-20, targets secretory proteins into two distinct intracellular pathways. When the DNA that encodes trypsinogen is introduced into AtT-20 cells, the protein is sorted into the regulated secretory pathway as efficiently as the endogenous peptide hormone ACTH. In this study we have used double-label immunoelectron microscopy to demonstrate that trypsinogen colocalizes in the same secretory granules as ACTH. In vitro mutagenesis was used to test whether the information for targeting trypsinogen to the secretory granules resides at the amino (NH2) terminus of the protein. Mutations were made in the DNA that encodes trypsinogen, and the mutant proteins were expressed in AtT-20 cells to determine whether intracellular targeting could be altered. Replacing the trypsinogen signal peptide with that of the kappa immunoglobulin light chain, a constitutively secreted protein, does not alter targeting to the regulated secretory pathway. In addition, deletion of the NH2 terminal "pro" sequence of trypsinogen has virtually no effect on protein targeting. However, this deletion does affect the signal peptidase cleavage site, and as a result the enzymatic activity of the truncated trypsin protein is abolished. We conclude that neither the signal peptide nor the 12 NH2-terminal amino acids of trypsinogen are essential for sorting to the regulated secretory pathway of AtT-20 cells. PMID- 3040771 TI - Reversible binding of actin to gelsolin and profilin in human platelet extracts. AB - This paper documents the reversible appearance of high-affinity complexes of profilin and gelsolin with actin in extracts of platelets undergoing activation and actin assembly. Sepharose beads coupled to either monoclonal anti-gelsolin antibodies or to polyproline were used to extract gelsolin and profilin, respectively, from EGTA-containing platelet extracts and determine the proportion of these molecules bound to actin with sufficient affinity to withstand dilution (high-affinity complexes). Resting platelets (incubated for 30 min at 37 degrees C after gel filtration) contained nearly no high-affinity actin/gelsolin or actin/profilin complexes. Thrombin, within seconds, caused quantitative conversion of platelet profilin and gelsolin to high-affinity complexes with actin, but these complexes were not present 5 min after stimulation. The calcium dependent actin filament-severing activity of platelet extracts, a function of free gelsolin, fell in concert with the formation of EGTA-stable actin/gelsolin complexes, and rose when the adsorption experiments indicated that free gelsolin was restored. The dissociation of high-affinity complexes was temporally correlated with the accumulation of actin in the Triton-insoluble cytoskeleton. PMID- 3040773 TI - Complete nucleotide sequence and deduced polypeptide sequence of a nonmuscle myosin heavy chain gene from Acanthamoeba: evidence of a hinge in the rodlike tail. AB - We have completely sequenced a gene encoding the heavy chain of myosin II, a nonmuscle myosin from the soil ameba Acanthamoeba castellanii. The gene spans 6 kb, is split by three small introns, and encodes a 1,509-residue heavy chain polypeptide. The positions of the three introns are largely conserved relative to characterized vertebrate and invertebrate muscle myosin genes. The deduced myosin II globular head amino acid sequence shows a high degree of similarity with the globular head sequences of the rat embryonic skeletal muscle and nematode unc 54 muscle myosins. By contrast, there is no unique way to align the deduced myosin II rod amino acid sequence with the rod sequence of these muscle myosins. Nevertheless, the periodicities of hydrophobic and charged residues in the myosin II rod sequence, which dictate the coiled-coil structure of the rod and its associations within the myosin filament, are very similar to those of the muscle myosins. We conclude that this ameba nonmuscle myosin shares with the muscle myosins of vertebrates and invertebrates an ancestral heavy chain gene. The low level of direct sequence similarity between the rod sequences of myosin II and muscle myosins probably reflects a general tolerance for residue changes in the rod domain (as long as the periodicities of hydrophobic and charged residues are largely maintained), the relative evolutionary "ages" of these myosins, and specific differences between the filament properties of myosin II and muscle myosins. Finally, sequence analysis and electron microscopy reveal the presence within the myosin II rodlike tail of a well-defined hinge region where sharp bending can occur. We speculate that this hinge may play a key role in mediating the effect of heavy chain phosphorylation on enzymatic activity. PMID- 3040772 TI - Changes in the expression of alpha-fodrin during embryonic development of Xenopus laevis. AB - Fodrin (nonerythroid spectrin) and its associated proteins have been previously implicated in the establishment of specialized membrane-cytoskeletal domains in differentiating cells. Using antiserum which is monospecific for the alpha subunit of fodrin, we demonstrate that alpha-fodrin is present in oocytes and adult tissues of Xenopus laevis. Analyses of the de novo synthesis of alpha fodrin during embryonic development reveal that alpha-fodrin is synthesized in oocytes, but not during early development. To investigate the level of control of alpha-fodrin expression, we isolated two cDNA clones for oocyte alpha-fodrin. The oocyte cDNA clones were identified as encoding portions of alpha-fodrin based on DNA sequence analysis and on the comparison of the predicted amino acid sequence of the cDNAs with the known sequence of human erythrocyte alpha-spectrin. The Xenopus alpha-fodrin cDNAs hybridize to a transcript of approximately 9 kb on RNA blots, and probably to a single gene type on genomic DNA blots. Both RNA blot analyses and S1 nuclease protection assays with the Xenopus alpha-fodrin cDNAs demonstrate that the observed decline in the de novo synthesis of alpha-fodrin polypeptides is controlled by a dramatic decrease in the abundance of alpha fodrin transcripts after fertilization. In contrast, levels of actin transcripts do not decrease during this period. Inasmuch as steady-state levels of alpha fodrin transcripts rise by the neurula stage of development, these results suggest that the synthesis of alpha-fodrin polypeptides during embryonic development of Xenopus is regulated, rather than constitutive, and that the primary level of control is the steady-state abundance of mRNA. PMID- 3040774 TI - Mouse ovarian granulosa cells produce urokinase-type plasminogen activator, whereas the corresponding rat cells produce tissue-type plasminogen activator. AB - It is well established that rat ovarian granulosa cells produce tissue plasminogen activator (tPA). The synthesis and secretion of the enzyme are induced by gonadotropins, and correlate well with the time of follicular rupture in vivo. We have found that in contrast, mouse granulosa cells produce a different form of plasminogen activator, the urokinase-type (uPA). As with tPA synthesis in the rat, uPA production by mouse granulosa cells is induced by gonadotropins, dibutyryl cAMP, and prostaglandin E2. However, dexamethasone, a drug which has no effect on tPA synthesis in rat cells inhibits uPA synthesis in the mouse. Results of these determinations made in cell culture were corroborated by examining follicular fluid, which is secreted in vivo predominantly by granulosa cells, from stimulated rat and mouse ovarian follicles. Rat follicular fluid contained only tPA, and mouse follicular fluid only uPA, indicating that in vivo, granulosa cells from the two species are secreting different enzymes. The difference in the type of plasminogen activator produced by the rat and mouse granulosa cells was confirmed at the messenger RNA level. After hormone stimulation, only tPA mRNA was present in rat cells, whereas only uPA mRNA was found in mouse cells. Furthermore, the regulation of uPA levels in mouse cells occurs via transient modulation of steady-state levels of mRNA, a pattern similar to that seen with tPA in rat cells. PMID- 3040776 TI - Fibronectin stimulates growth but not follicle-stimulating hormone-dependent differentiation of rat granulosa cells in vitro. AB - Since fibronectin is a secretory product of immature rat granulosa cells in culture and may contribute to the follicular microenvironment in vivo, we have studied the effects of this adhesion factor on follicle-stimulating hormone (FSH) dependent differentiation in short-term (2-3-day) cultures and on growth and protein synthesis in long-term (12-day) cultures. In comparison with cells plated on tissue culture plastic, those plated on an optimal fibronectin-coated substratum showed much greater cell spreading. There were no short-term effects of this morphological change on FSH-stimulation of cyclic AMP production, apparent activities of aromatase or cholesterol side-chain cleavage enzymes, or acquisition of luteinizing hormone (LH) responsiveness in cultured cells. However, progesterone metabolism to 20 alpha-hydroxypregnan-4-en-3-one was increased. Only cultures on fibronectin showed increases between days 3 and 9 in protein (2.5-fold) and DNA (1.4-fold) contents. Cells cultured on fibronectin also showed greater uptake and incorporation of [3H]leucine in comparison with cells cultured on plastic. FSH treatment caused cell aggregation and rounding and delayed the increase in protein content of cells cultured on fibronectin. The results presented demonstrate that the principal direct effect of fibronectin mediated adhesion on rat granulosa cells is to enhance cell maintenance and growth, while having no generalized action on FSH-dependent differentiation. PMID- 3040775 TI - Multiple carbohydrate receptors on lymphocytes revealed by adhesion to immobilized polysaccharides. AB - Phosphomannan polysaccharides and fucoidan, a polymer of fucose 4-sulfate, have been demonstrated to inhibit adhesion of lymphocytes to tissue sections that contain high endothelial venules (Stoolman, L. M., T. S. Tenforde, and S. D. Rosen, 1984, J. Cell Biol., 99:1535-1540). We have investigated the potential cell surface carbohydrate receptors involved by quantitating adhesion of rat cervical lymph node lymphocytes to purified polysaccharides immobilized on otherwise inert polyacrylamide gels. One-sixth of the lymphocytes adhered specifically to surfaces derivatized with PPME (a phosphomannan polysaccharide prepared from Hansenula holstii yeast), whereas up to half of the cells adhered to surfaces derivatized with fucoidan. Several lines of evidence demonstrated that two distinct receptors were involved. Adhesion to PPME-derivatized gels was labile at 37 degrees C (decreasing to background levels within 120 min) whereas adhesion to fucoidan-derivatized gels was stable. Soluble PPME and other phosphomannans blocked adhesion only to PPME-derivatized gels; fucoidan and a structurally related fucan blocked adhesion to fucoidan-derivatized gels. Other highly charged anionic polysaccharides, such as heparin, did not block adhesion to either polysaccharide-derivatized gel. Adhesion to PPME-derivatized gels was dependent on divalent cations, whereas that to fucoidan-derivatized gels was not. The PPME-adherent lymphocytes were shown to be a subpopulation of the fucoidan adhesive lymphocytes which contained both saccharide receptors. These data reveal that at least two distinct carbohydrate receptors can be found on peripheral lymphocytes. PMID- 3040777 TI - Dexamethasone inhibits feedback regulation of the mitogenic activity of tumor necrosis factor, interleukin-1, and epidermal growth factor in human fibroblasts. AB - Tumor necrosis factor (TNF), interleukin-1 (IL-1), and epidermal growth factor (EGF) were mitogenic for human diploid FS-4 fibroblasts. Dexamethasone amplified the growth-stimulating action of all three agents. Amplification of the growth stimulating action was maximal when dexamethasone was added along with TNF or EGF; no amplification was seen if the addition of dexamethasone was delayed for more than 3 hr. Prolonged simultaneous treatment with TNF and EGF resulted in less growth stimulation than treatment with EGF alone. Dexamethasone abolished this apparent antagonistic interaction between TNF and EGF. Dexamethasone also inhibited the antiviral action of TNF against encephalomyocarditis (EMC) virus in FS-4 cells. TNF and IL-1 increased the steady state level of interferon (IFN) beta 2 mRNA but failed to induce detectable levels of IFN-beta 1 mRNA in FS-4 cells. Dexamethasone inhibited the increase of IFN-beta 2 mRNA levels by IL-1 or TNF. Inhibition of IFN-beta synthesis is likely to be responsible for the inhibition of the TNF-induced antiviral state by dexamethasone. Since IFNs suppress cell growth, inhibition of endogenous IFN-beta synthesis may also be responsible for the amplification by dexamethasone of the growth-stimulating action of TNF and IL-1. Amplification of the mitogenic action of EGF by dexamethasone appears to be mediated by different mechanism. PMID- 3040778 TI - The effect of factors released from the tumor-transformed cells on DNA synthesis, mitosis, and cellular enlargement in 3T3 fibroblasts. AB - Quiescent serum-starved 3T3 cells can be stimulated to initiate DNA synthesis after addition of conditioned media from spontaneously tumor-transformed 3T3 cells (3T6-cells) or from SV-40-transformed 3T3 cells (SV-3T3 cells). The conditioned media were found to stimulate both the chromosome cycle (i.e., DNA synthesis and cell division) and the growth cycle (i.e., cellular enlargement). Furthermore, addition of conditioned media to quiescent 3T3 cells increased the activity of HMG CoA reductase--an enzyme previously proposed to exercise some control on cell proliferation in 3T3 cells (Larsson and Zetterberg: J. Cell. Physiol. 129:99-102, 1986. The increased activity of HMG CoA reductase after treatment with tumor cell conditioned media was correlated to the stimulatory effects on DNA synthesis. By treating 3T3 cells stimulated to resume proliferation by addition of conditioned media with mevinolin (a competitive inhibitor of HMG CoA reductase) the activity of HMG CoA reductase as well as the DNA synthesis and cell division were efficiently inhibited. In contrast, HMG CoA activity was not coupled to the cellular enlargement. Therefore, it is proposed that one set of factors present in tumor cell conditioned media preferentially stimulates the chromosome cycle by increasing the HMG-CoA reductase activity, whereas another set of factors is responsible for growth in cell size. Both types of factors are required for balanced growth. PMID- 3040779 TI - Both protein kinase C and calcium mediate activation of the Na+/H+ antiporter in Chinese hamster embryo fibroblasts. AB - Chinese hamster embryo fibroblast cells (CHEF/18) possess a plasma membrane associated, amiloride-sensitive Na+/H+ antiporter that affects intracellular pH (pHi) and is activated by growth factor addition. Our results using 14C-benzoic acid distribution indicate that both epidermal growth factor (EGF) and thrombin are capable of causing rapid rises in the pHi of CHEF/18 cells. The maximal shift induced by these factors is 0.20 to 0.25 pH units above the basal unstimulated level. Distinctive differences were observed between the modes of action of these two growth factors. Sequential additions revealed that the rise in pHi due to EGF was additive with that caused by diacylglycerols (DAG), while that of thrombin was not. Furthermore, exposure of cells to the phorbol ester PMA for a prolonged period of time in order to down-regulate protein kinase C (pkC), or treatment with the pkC inhibitor H-7, abolished the pHi response to thrombin but not to EGF. In contrast, incubation of cells in nominally calcium-free medium or with the calmodulin antagonists W-7 or trifluoperazine (TFP) decreased only the ability of EGF to cause changes in pHi. These data suggest that there are two distinct mechanisms for activation of the Na+/H+ antiporter in CHEF/18 fibroblast cells and thus provide an example of the use of alternative modes for the modulation of intracellular processes. PMID- 3040780 TI - The glioma cell-derived neurite promoting activity protein is functionally and immunologically related to human protease nexin-I. AB - Protease nexin-I (PN-I, Mr approximately 43,000) is representative of a newly described class of cell-secreted protease inhibitors. PN-I has been purified to apparent homogeneity, partially sequenced, and monospecific antibodies have been raised against it. PN-I is a potent inhibitor of urokinase, thrombin, plasmin, and trypsin. In addition, cells have specific receptors that mediate the uptake of covalently linked complexes formed between PN-I and its protease substrates. In the present studies, we have investigated the relationship between human PN-I and a protease inhibitor derived from C6 glioma cells in culture that has neurite promoting activity. On the basis of co-purification on heparin-Sepharose, identical molecular weight, antibody cross-reactivity, and receptor cross reactivity, we conclude that PN-I and the glioma-cell-derived inhibitor are equivalent molecules. PMID- 3040781 TI - Serum-activated T51B rat liver cells transiently accumulate cyclic AMP-dependent protein kinases on their surfaces during the G1 phase. AB - Confluent T51B rat liver epithelial cells promptly began accumulating cyclic AMP binding sites on their surfaces when they were stimulated from quiescence by serum growth factors in medium containing 1.8 mM Ca2+, but they began losing the accumulated binding sites shortly before initiating DNA replication. When the medium contained only 0.02 mM Ca2+, the cells still accumulated surface cyclic AMP-binding sites, but they did not initiate DNA replication and tended to continue accumulating the binding sites. The cyclic AMP-binding sites were eliminated completely by treating intact cells for 5 minutes with 0.005% trypsin (which did not damage the cells), and cyclic AMP caused them to be released from intact, undamaged cells into the medium. The binding sites also comigrated electrophoretically with purified regulatory subunits of type I cyclic AMP dependent protein kinase, and to a lesser extent the regulatory subunit of type II cyclic AMP-dependent protein kinase. Therefore, it is likely that a transient accumulation of cyclic AMP-dependent protein kinases on the outer surface of the plasma membrane is part of the T51B rat liver cell's prereplicate program. PMID- 3040782 TI - Human iliac artery endothelial cells express both genes encoding the chains of platelet-derived growth factor (PDGF) and synthesize PDGF-like mitogen. AB - In human umbilical vein and bovine aortic endothelial cells in culture c-sis gene expression and secretion of platelet-derived growth factor (PDGF) has been previously demonstrated. We now report the presence of PDGF-1 and PDGF-2/sis mRNA transcripts in primary cultures of human iliac artery endothelial cells (HIA-EC). Concomitantly, these cells synthesize and secrete PDGF-like proteins identified by direct immunoprecipitation with specific PDGF antiserum. The PDGF proteins secreted by HIA-EC have molecular weights of 31 and 35 kd under nonreducing conditions. Upon reduction these proteins are converted to the monomeric 15- and 16-kd forms. Conditioned media derived from HIA-EC stimulated the incorporation of 3H-thymidine by 3T3 cells and competed with 125I-PDGF for its binding to 3T3 cell membrane receptors. The biologic activity was stable to heating at 100 degrees C for 10 min and sensitive to reducing agents, properties similar to those of authentic PDGF. Production of PDGF-like mitogen by the human arterial endothelial cells may play an important role in the paracrine modulation of arterial wall regeneration following vascular injury. PMID- 3040783 TI - [Esophageal Abrikosov's tumor, preoperative diagnosis with echoendoscopy]. AB - Echo-endoscopic investigation provided the preoperative diagnosis in a case of oesophageal Abrikossoff's tumor. Characteristics of this rare tumor are described, and difficulties of etiologic diagnosis prior to histology underlined, using conventional exploratory examinations. Minimal surgery should be attempted for this exceptionally malignant tumor. PMID- 3040784 TI - [Spontaneous hemoperitoneum caused by rupture of a hepatocarcinoma]. PMID- 3040785 TI - Paraneoplastic syndromes. AB - As the lengthy but nonetheless incomplete review suggests, paraneoplastic syndromes are protean in their manifestations and, for the most part, poorly understood. Indeed, some of the more common abnormalities in cancer patients that might be considered paraneoplastic--such as anorexia-cachexia syndrome or unexplained fever--have not been discussed because they are so poorly understood. Most of the syndromes reviewed are either clearly paraneoplastic or strongly associated with cancer. Their clinical importance does not lie in the number of patients affected; it is a small minority. Instead, the syndromes may occasionally be helpful in the diagnosis of cancer or in monitoring response to cancer therapy. They may also be confused with the effects of metastatic disease. In some patients, amelioration of the syndromes can reverse the patient's dominant symptoms and thus provide significant clinical palliation. In a more general context, studies of etiologic mechanisms in paraneoplastic syndromes may offer insights into a variety of unexplained abnormalities in cancer patients. The best-understood syndromes result from tumor production of biologically active substances or, to a lesser extent, from autoimmune phenomena. These would appear to be probable mechanisms in many recognized paraneoplastic syndromes of uncertain etiology and perhaps in some heretofore unrecognized paraneoplastic syndromes. Finally, paraneoplastic syndromes could also hold clues to the neoplastic process. Better understanding of the ways in which tumors regulate remote effects--such as release of TGFs--may ultimately enhance our knowledge of tumor growth itself. PMID- 3040786 TI - Toward a national AIDS prevention strategy. PMID- 3040787 TI - Back pain and a nonproductive cough. PMID- 3040788 TI - Gene transplants into germ cells. PMID- 3040789 TI - Argentation liquid chromatography of polynuclear aromatic hydrocarbons on a silver(I)-loaded mercaptopropyl silica gel stationary phase. PMID- 3040790 TI - Liquid chromatographic retention behaviour in the separation of anti-epileptic drugs on phenyl-, diphenyl-, and triphenyl-bonded silicas and glasses. AB - The retention and selectivity behaviour of some anti-epileptic drugs were studied by high-performance liquid chromatography on 21 kinds of phenyl-modified porous glasses and silicas, prepared from solutions of phenyldimethylchlorosilane, diphenylmethylchlorosilane or triphenylchlorosilane in xylene, and from various kinds of glass or silica with various mean pore diameters and/or specific surface areas. From elemental analysis data for carbon, the maximum number of bonded phenyl surface groups per gram (mean pore diameter 15 nm, specific surface area 217 m2/g, pore volume 0.85 ml/g) in phenyl-, diphenyl-, and triphenyl-bonded gels was calculated to be 0.313, 0.159, and 0.112 X 10(21), respectively. Using various acetonitrile-0.01 M potassium dihydrogen phosphate mixtures as eluents, the anti-epileptic drugs were separated on all the gels studied, but with different degrees of resolution. With increase in specific surface area on the glasses or silicas, the k' values of three anti-epileptic drugs increased. The selectivity for the separation of carbamazepine and diphenylhydantoin is discussed and explained by the pi-pi interaction between solutes and stationary phases. It has been shown that diphenyl and triphenyl phases are more suitable stationary phases for the selective separation of anti-epileptic drugs than monophenyl phases. PMID- 3040791 TI - Gradient liquid chromatographic method for the simultaneous determination of sweeteners, preservatives and colours in soft drinks. PMID- 3040792 TI - Zonal high-performance affinity chromatography as a tool for protein interaction studies with special reference to the lipase-colipase complex. AB - The technique of zonal high-performance affinity chromatography applied to the lipase-colipase system (lipase B as eluted acceptor and colipase as silica-bonded ligand) gave qualitatively the same results as conventional affinity chromatography. The elution volume of the acceptor increases with decreasing load introduced at constant volume into the column of ligand-bonded silica. This led to the use of a mathematical treatment for calculating the dissociation constant (KD) of the lipase-colipase complex. The influence of some physical and chemical chromatographic parameters was studied. Increasing temperature and flow-rate reduced the affinity of lipase for colipase, whereas it was only slightly modified by increasing the ionic strength. The KD value was minimal and equal to 0.1 X 10(-6) M at pH 4.7 and 0.38 X 10(-6) M at pH 6.5, after correction for the flow-rate. The latter value is similar to that obtained by more conventional techniques. The absence of some marked KD modifications by ionic strength and the value of delta S for the complex association obtained by temperature studies suggest the intervention of mixed hydrophobic-ionic interactions in the formation of the lipase-colipase complex. Their respective importances are discussed. PMID- 3040793 TI - Retention behaviour of phenylthiohydantoin amino acids in micro high-performance liquid chromatography with octadecyl bonded glasses and silicas. PMID- 3040794 TI - Pharmacognostical studies of Tabernaemontana species. XX. Ion-pair droplet counter-current chromatography of indole alkaloids from suspension cultures. PMID- 3040795 TI - Chromatographic separation of DNA restriction fragments. AB - Reversed-phase liquid chromatography (RPC-5) was the first chromatographic technique to be successfully applied to DNA restriction fragments. Size-exclusion high-performance liquid chromatography (HPLC) has the advantage of wide variability in buffer conditions but is restricted to an upper size limit at 800 base pairs. Anion-exchange HPLC is most versatile and may best be scaled up for the preparation of milligram amounts of specific DNA fragments. The preparation of the sample before and after chromatography is described. Because several applications of DNA fragments will be complemented by RNA fragments, their preparation and purification is included. Disposable small-scale columns are available for processing many samples. Future medical applications of HPLC of DNA fragments concern medical microbiology, human genetics and forensic medicine. PMID- 3040796 TI - Adrenocortical function in acquired immunodeficiency syndrome. AB - Clinical features of adrenal steroid deficiency occur in patients with the acquired immunodeficiency syndrome (AIDS). To determine the frequency of aberrations in peripheral steroid levels in patients with AIDS and AIDS-related complex (ARC) we measured morning recumbent plasma cortisol, deoxycorticosterone, 18-hydroxydeoxycorticosterone (18-OHDOC), corticosterone, aldosterone, and 18 hydroxycorticosterone concentrations before and after administration of 0.25 mg ACTH (Cosyntropin) in 74 randomly selected hospitalized patients with AIDS and 19 patients with ARC. Basal (0800 h) cortisol levels in the AIDS patients were significantly higher (P less than 0.01) than those in normal subjects, while other ACTH-dependent steroids of the 17-deoxypathway, deoxycorticosterone, corticosterone, and 18-OHDOC, were normal. These latter steroids increased subnormally in response to ACTH in patients with either AIDS (P less than 0.001) or ARC (P less than 0.005), but in ARC patients plasma 18-OHDOC levels were significantly higher than in those with AIDS (P less than 0.001). Supraphysiological doses of ACTH were then administered for 3 consecutive days to 14 patients with AIDS and 9 with ARC, which confirmed and amplified the subnormal responses of these steroids in AIDS. The mean plasma cortisol response was reduced on the third day only in AIDS patients, whereas in the ARC patients the steroid responses were normal. Angiotensin III infusion and postural stimulation increased plasma aldosterone and 18-hydroxycorticosterone levels in AIDS and ARC patients. Defective stimulation of 18-OHDOC alone or in combination with defective stimulation of other 17-deoxysteroids can be a harbinger of subsequent impaired adrenal capacity in AIDS. PMID- 3040797 TI - Normal suppressive T cell function of Epstein-Barr virus-induced B cell activation in Graves' disease. AB - Several studies have demonstrated abnormalities of T cell regulation of Epstein Barr virus (EBV)-induced B cell activation in systemic autoimmune diseases, such as rheumatoid arthritis, systemic lupus erythematosus, and systemic sclerosis. To investigate whether this abnormality is a common feature of other autoimmune diseases, we studied 10 EBV-immune normal subjects and 22 EBV-immune patients with Graves' disease (GD); 11 had newly diagnosed hyperthyroidism, and 11 had received carbimazole treatment for hyperthyroidism for at least 6 months. Peripheral B lymphocytes infected with EBV were cultured for 20 days in the presence or absence of autologous T cells at different ratios. Immunoglobulins M and G secretion into the supernatants was determined using an enzyme-linked immunosorbent assay. The extent of suppression when T cells were added, as measured by a suppression ratio, was not significantly different in normal subjects and newly diagnosed GD patients (0.65 vs. 0.63 on the 16th day and 0.77 vs. 0.72 on the 20th day of culture, respectively). In carbimazole-treated patients, the appearance of functional suppressor T cells was delayed slightly, but the overall suppression ratios on the 16th and 20th days were normal. Thus, a T cell regulation abnormality of EBV-induced B cell activation could not be demonstrated in patients with untreated hyperthyroid GD, suggesting that the autoimmune reactivity in such patients is probably dependent upon a specific thyroid suppression defect rather than a generalized suppression defect. PMID- 3040798 TI - Comparison of methods for detection of serum antibody to murine rotavirus. AB - Mice are frequently used as animal models for the study of rotaviral infections. Since natural infection is common in laboratory mice, it is important that rotaviral studies, as well as other studies utilizing suckling mice, employ animals of known immune status to murine rotavirus. A variety of homologous and heterologous enzyme immunoassay systems and an immunofluorescence technique were thus compared to determine the immunoassay that is most effective at detecting adult mice seropositive for rotaviral antibody. It was determined that a homologous enzyme immunoassay inhibition technique utilizing murine rotavirus derived reagents was the most efficient serologic assay evaluated. A serologic response was consistently detected by this assay by 5 days after experimental rotaviral inoculation of adult mice. A homologous antibody-binding enzyme immunoassay, a heterologous inhibition enzyme immunoassay utilizing antigenically related simian rotavirus (SA-11) reagents, and an immunofluorescence technique utilizing Nebraska calf diarrhea virus antigens were found to be less sensitive for detecting serum antibody to murine rotavirus. PMID- 3040799 TI - Subclass restriction of human enterovirus antibodies. AB - We studied antibodies to enteroviruses in four groups of serum specimens: those from healthy adults, cord blood specimens, serum specimens known to contain immunoglobulin M (IgM) to coxsackie B (CB) viruses by radioimmunoassay, and serum specimens from children with symptomatic enteroviral infections. Enzyme-linked immunosorbent assays (ELISAs) were developed to detect the IgG class- and subclass (IgG1, IgG2, IgG3, and IgG4)-specific responses to CB3. The CB3 virus ELISA was not type specific. There was very poor correlation between CB3 virus neutralizing titer and IgG anti-CB3 virus ELISA results, indicating that antibodies to heterologous picornaviruses cross-react with CB3 virus in the assay. All serum specimens tested except one were IgG positive for CB3 virus. All 32 cord serum specimens were positive for IgG1 and IgG3. No enterovirus-specific IgG2 or IgG4 was detected in any serum specimen tested. Most serum specimens from the IgM-positive group, healthy adults, and children with enterovirus infections were positive for IgG1 and IgG3. Class and subclass antibody titers remained constant over time. IgG antibodies to enteroviruses appear to be restricted to the IgG1 and IgG3 subclasses. This pattern is similar to results obtained by other investigators evaluating IgG subclass antibodies to protein antigens. PMID- 3040800 TI - Impact of cell culture sensitivity and virus concentration on rapid detection of herpes simplex virus by cytopathic effects and immunoperoxidase staining. AB - Tremendous interest has been generated in the commercial kits now available that incorporate herpes simplex virus isolation in cell culture with immunoperoxidase staining for viral antigen detection. Most studies comparing commercial kits with conventional cell culture techniques have found the kits to be less sensitive. However, different cell cultures were used for the two methods. In this study, mink lung, rabbit kidney, MRC-5, and Vero cells were compared for reisolation of herpes simplex virus from clinical specimens in which viral infectivity titers were concurrently determined. When specimens contained high titers of infectious virus, the cell system used made little difference and all specimens were detected by immunoperoxidase staining at 48 h postinoculation. However, when specimens contained low concentrations of virus, the differences in sensitivity between cell systems became apparent in rapidity of detection and overall isolation rate. Mink lung and rabbit kidney cells were both more sensitive than MRC-5 cells; Vero cells were significantly less sensitive than the other cells tested. The application of immunoperoxidase staining shortened the time to virus detection and lessened, but did not eliminate, the differences between the cell systems. Cytopathic effects alone in the most sensitive cell system equaled or exceeded immunoperoxidase staining applied in less-sensitive cell cultures. PMID- 3040801 TI - Improvement of serological diagnosis of neonatal cytomegalovirus infection by simultaneously testing for specific immunoglobulins E and M by antibody-capture enzyme-linked immunosorbent assay. AB - This study describes the results of testing 92 serum samples, including 10 umbilical cord serum samples, from 38 cytomegalovirus (CMV)-infected neonates for CMV-specific immunoglobulins E (IgE) and M by antibody-capture enzyme-linked immunosorbent assays with enzyme-labeled CMV antigen. All infants excreted CMV in the urine. It was demonstrated that the CMV IgE test was more sensitive than the CMV IgM test in diagnosing CMV infection in neonates by serology. Thus, the sensitivity for the IgE test was 82%, whereas for the IgM test it was only 66%. Furthermore, the CMV-specific IgE response, expressed as absorbance, was higher than the specific IgM response in 91% of the 76 sera which contained antibodies of either one or both immunoglobulin classes. Forty-six control sera, including 18 umbilical cord sera, from 46 neonates from whom CMV was not isolated were also tested. Most sera were negative. Three infants, however, had CMV antibodies of one or both classes, indicating infection. The level of total serum IgE was controlled in 24 of the sera from the CMV-infected neonates, but in none of the cases was the level elevated. No correlation was found between the reactivity of the antibodies in the two immunoglobulin classes and the level of CMV in urine. PMID- 3040802 TI - Isolation and analysis of restriction endonuclease digestive patterns of chromosomal DNA from Mycobacterium paratuberculosis and other Mycobacterium species. AB - A relatively rapid and efficient method for the extraction of chromosomal DNA from Mycobacterium paratuberculosis and other mycobacteria was developed. Approximately 25 to 50 micrograms of DNA could be extracted from 100 mg (wet weight) of cells, which was sufficient to perform several restriction endonuclease analyses from a single preparation. The DNA from five Mycobacterium species, including four strains of M. paratuberculosis and four strains of M. avium, was analyzed by this method. Digestion with the restriction endonucleases BstEII and PstI yielded the most definitive restriction patterns. For some strains, the restriction endonuclease analysis results were in agreement with the current identification of these organisms. The two strains of M. avium serotype 2 had identical fragment patterns. Similarly, the two strains of M. avium complex serotype 6 had identical fragment patterns. The three mycobactin-dependent M. paratuberculosis strains were very similar, whereas the mycobactin-independent M. paratuberculosis strain was more similar to the M. avium serotype 2 strains. Although many more cultures would need to be evaluated to determine correct groupings, the results of this study demonstrated the potential of restriction enzyme analysis for the differentiation of slowly growing mycobacteria. PMID- 3040804 TI - Use of somatosensory evoked potentials to evaluate the peripheral nervous system. AB - Somatosensory evoked potentials (SEPs) are being used increasingly to evaluate peripheral somatosensory pathways. They have been used in patients with plexus lesions but may provide misleading information when multiple lesions are present, demonstrating only the electrophysiologic consequences of the most distal lesion that is present. Ulnar SEPs can be abnormal in neurogenic thoracic outlet syndrome, whereas they are normal in the nonneurogenic variety. SEPs to nerve trunk stimulation are generally not helpful in patients with isolated radiculopathies, and SEPs elicited by dermatomal or cutaneous nerve stimulation have provided conflicting results. SEPs may be important in evaluating conduction along inaccessible proximal segments of limb nerves. Their value in Guillain Barre syndrome, in which pathology may be predominantly proximal, is unclear. In evaluating peripheral neuropathies, SEPs can sometimes be useful when peripheral sensory nerve action potentials are unobtainable; sometimes, however, SEPs provide misleading information concerning conduction velocity. The SEP findings should not be relied on to determine whether sensory loss is organic or nonorganic, although they are one factor to consider in making this determination. PMID- 3040803 TI - Evaluation of a direct fluorescein-conjugated monoclonal antibody for detection of cytomegalovirus in centrifugation culture. AB - A fluorescein-conjugated murine monoclonal antibody (MAb) reactive with cytomegalovirus (CMV) was evaluated for the detection of CMV in centrifugation culture. Of 188 specimens, 90 were positive for CMV in centrifugation culture. The fluorescein-conjugated MAb detected CMV in 86 of 90 (95%) specimens at 16 h postinoculation, and 88 of 90 (98%) were positive at 36 h. The fluorescein conjugated MAb can be used in a direct immunofluorescence assay that can be completed in 15 min following cover slip fixation. Use of this antibody in centrifugation culture provides a convenient and rapid assay for the identification of CMV. PMID- 3040805 TI - Ursodeoxycholate stimulates Na+-H+ exchange in rat liver basolateral plasma membrane vesicles. AB - Na+:H+ and Cl-:HCO3- exchange are localized, respectively, to basolateral (blLPM) and canalicular (cLPM) rat liver plasma membranes. To determine whether these exchangers play a role in bile formation, we examined the effect of a choleretic agent, ursodeoxycholate (UDCA), on these exchange mechanisms. 22Na (1 mM) and 36Cl (5 mM) uptake was determined using outwardly directed H+ and HCO3- gradients, respectively. Preincubation of blLPM vesicles with UDCA (0-500 microM) resulted in a concentration-dependent increase in initial rates of amiloride sensitive pH-driven Na+ uptake, with a maximal effect at 200 microM. UDCA (200 microM) increased Vmax from 23 +/- 2 (control) to 37 +/- 7 nmol/min per mg protein; apparent Km for Na+ was unchanged. Preincubation with tauroursodeoxycholate (200 microM), taurocholate (10-200 microM) or cholate, chenodeoxycholate, or deoxycholate (200 microM) had no effect on pH-driven Na+ uptake. UDCA (200 microM) had no effect on either membrane lipid fluidity, assessed by steady-state fluorescence polarization using the probes 1,6-diphenyl 1,3,5-hexatriene, 12-(9-anthroyloxy) stearic acid, and 2-(9-anthroyloxy) stearic acid (2-AS), or Na+,K+-ATPase activity in blLPM vesicles. In cLPM vesicles, UDCA (0-500 microM) had no stimulatory effect on initial rates of HCO3(-)-driven Cl- uptake. Enhanced basolateral Na+:H+ exchange activity, leading to intracellular HCO3- concentrations above equilibrium, may account for the bicarbonate-rich choleresis after UDCA infusion. PMID- 3040806 TI - Mechanism of increased alpha adrenergic vasoconstriction in human essential hypertension. AB - Multiple components of vascular alpha adrenergic responsiveness were investigated in twenty-four men with mild hypertension and eighteen age- and weight-matched normotensive controls. Arterial plasma norepinephrine (paNE), an index of sympathetic drive, was increased in hypertensives compared to normotensives (mean +/- SE), 199 +/- 24 vs. 134 +/- 11 pg/ml, P less than 0.02. The effective concentration of intra-arterial (iaNE) increasing forearm vascular resistance (FAVR) 30% (NE-EC30, an index of vascular alpha-receptor sensitivity) was similar in normotensives and hypertensives, 9 +/- 1 vs. 13 +/- 3 ng/100 ml per min, respectively, P greater than 0.3. The phentolamine induced reduction in FAVR, an index of vascular alpha-tone, was greater in hypertensives, -21.3 +/- 1.8 vs. normotensives, -14.9 +/- 1.2 U, P less than 0.02. We interpret these data as evidence for normal vascular alpha-receptor sensitivity to norepinephrine in mild hypertensives. Consequently, the increased sympathetic drive in mild hypertensives explains the elevated vascular alpha-tone. Although vascular alpha receptor sensitivity to iaNE was normal, the FAVR responses at high doses (reactivity) were greater in hypertensives to regional infusion of both NE and angiotensin II. This "nonspecific" enhancement of vascular reactivity is probably explained by structural vascular changes in hypertensives. PMID- 3040807 TI - Opioid-mediated suppression of interferon-gamma production by cultured peripheral blood mononuclear cells. AB - Mounting evidence suggests that opiate addiction and stress are associated with impaired cell-mediated immunity. We tested the hypothesis that morphine and the endogenous opioid beta-endorphin (beta-END), a pituitary peptide released in increased concentrations during stress, can suppress the production of the key macrophage-activating lymphokine interferon-gamma (IFN-gamma) by cultured human peripheral blood mononuclear cells (PBMNC). Using a radioimmunoassay to measure IFN-gamma, we found that exposure of PBMNC to biologically relevant concentrations of both opioids significantly inhibited IFN-gamma generation by cells stimulated with concanavalin A and varicella zoster virus. Studies of the mechanism of suppression revealed (a) a classical opioid receptor is involved (suppression was antagonized by naloxone and was specific for the NH2 terminus of beta-END), (b) monocytes are the primary target cell for opioids (monocyte depleted lymphocyte preparations showed little suppression), and (c) reactive oxygen intermediates (ROI) and prostaglandin E2 are important mediators (scavengers of ROI and indomethacin eliminated the suppression). Based on these findings we suggest that opioid-triggered release of inhibitory monocyte metabolites may play a role in the immunodeficiency associated with narcotic addiction and stress. PMID- 3040809 TI - Angiotensin-converting enzyme labeled with [3H]captopril. Tissue localizations and changes in different models of hypertension in the rat. AB - In vitro autoradiography with [3H]captopril was used to localize and quantitate angiotensin-converting enzyme (ACE) in various tissues in two-kidney, one-clip (2K-1C) hypertension, one-kidney, one-clip (1K-1C) hypertension, desoxycorticosterone acetate (DOCA)-salt hypertension, and a normotensive control group. There were no significant differences in mean systolic blood pressure among the hypertensive groups. Plasma renin activity (PRA) was highest in the 2K 1C group (6.20 +/- 2.17 ng/ml per h), intermediate in the 1K-1C group (2.19 +/- 0.62 ng/ml per h) and control group (3.20 +/- 0.53 ng/ml per h), and lowest in the DOCA-salt group (0.07 +/- 0.06 ng/ml per h). In the lungs, aorta, mesenteric arteries, and adrenal medulla, ACE labeling was highest in the 2K-1C group, intermediate in the 1K-1C and control groups, and lowest in the DOCA-salt group. ACE levels in these tissues correlated positively with PRA. In the kidney, anterior pituitary, testis, and choroid plexus of the brain, ACE levels correlated negatively with PRA, with lowest ACE levels in the 2K-1C group and highest levels in the DOCA-salt group. In the epididymis, posterior pituitary, and other regions of the brain, ACE levels did not differ significantly among the groups. PMID- 3040808 TI - Vasodilatory actions of alpha-human atrial natriuretic peptide and high Ca2+ effects in normal man. AB - To study vascular actions of synthetic alpha-human atrial natriuretic polypeptide (alpha hANP) in man, forearm blood flow (FBF) was measured by strain-gauge plethysmograph during the continuous infusion of 100 ng/min alpha hANP dissolved in 5% dextrose into the brachial artery in healthy subjects. alpha hANP increased FBF, with the concomitant increase in ipsilateral limb venous plasma concentrations of alpha hANP. Overall, there was a significant linear correlation between the decrements of ipsilateral forearm vascular resistance (FVR) during infusions of alpha hANP and initial FVR levels (r = -0.883, P less than 0.01). Moreover, alpha hANP, at the stepwise increasing doses of 20, 100, and 500 ng/min, increased FBF in a dose-related fashion: alpha hANP elicits a concentration-dependent vasodilation of forearm vascular beds. Concomitantly, infusions of alpha hANP caused a dose-dependent increase in ipsilateral limb venous plasma cyclic guanosine monophosphate (cyclic GMP). Overall, there were direct correlations of FBF either to ipsilateral venous plasma alpha hANP (r = 0.724, P less than 0.01) or to cyclic GMP concentrations (r = 0.637, P less than 0.01). Subsequently, isoosmolar CaCl2 solution was infused into the same brachial artery at a rate of 0.09 meq/min, and then, with a 2.5 +/- 0.2-mg/dl increase in ipsilateral venous serum calcium concentrations the incremental responses of both FBF and plasma cyclic GMP to alpha hANP were severely blunted. There was also a significant positive linear correlation between FBF and venous plasma cyclic GMP during infusions of alpha hANP with the simultaneous administration of CaCl2 (r = 0.807, P less than 0.01). Finally, the addition of CaCl2 infusion did not change the slope of the regression line of the FBF-plasma cyclic GMP relationship during infusions of alpha hANP. Evidence presented suggests that alpha hANP acts directly on the forearm vascular beds in man, eliciting its vascular relaxant effect, possibly by increasing cellular levels of cyclic GMP. Moreover, modest elevations of serum calcium inhibit the alpha hANP-dependent vasodilation, possibly through the suppression of cyclic GMP activation. PMID- 3040810 TI - Chronic glucocorticoid therapy amplifies glomerular injury in rats with renal ablation. AB - Functional and/or structural measurements were performed in eight groups of Munich-Wistar rats after five-sixths nephrectomy. Groups 1 and 5 received no therapy. Groups 2 and 6 received daily doses of methylprednisolone (MP). Groups 3 and 7 received MP plus the angiotensin I converting enzyme inhibitor (CEI), benzazepril. Groups 4 and 8 received CEI alone. Groups 1 through 4 underwent micropuncture study 2 wk after renal ablation. Untreated group 1 rats exhibited systemic hypertension and elevation of the single nephron glomerular filtration rate due to glomerular capillary hyperperfusion and hypertension. Administration of MP in group 2 resulted in comparable systemic hypertension, with further elevation of the single nephron glomerular filtration rate due to even higher values for glomerular perfusion and hydraulic pressure. Concurrent treatment with CEI in groups 3 and 4 controlled systemic and glomerular hypertension despite equivalent renal ablation and, in group 3, comparable doses of MP. Groups 5 through 8 were followed for 12 wk. Untreated group 1 rats demonstrated continued systemic hypertension, progressive proteinuria, and eventual glomerular sclerosis. Addition of MP in group 6 dramatically accelerated the development of proteinuria and glomerular sclerosis, while CEI (groups 7 and 8) afforded striking protection against disease progression. Thus, potent vasodilator glucocorticoids may amplify hemodynamically mediated glomerular injury, whereas control of systemic and glomerular hypertension prevents this undesirable consequence of chronic steroid therapy. PMID- 3040811 TI - Application of chromosomal restriction endonuclease digest analysis for use as typing method for Clostridium difficile. AB - The usefulness of restriction endonuclease analysis of chromosomal DNA as a typing method for Clostridium difficile was tested. Over four months all faecal samples were routinely cultured for C difficile. DNA of all isolated strains was isolated and tested with the restriction endonuclease Hind III. The patterns obtained after electrophoresis in agarose gels seemed to be strain specific. Antibiotic susceptibility profiles agreed with the results of the restriction endonuclease analysis, though they were much less discriminating. Analysis of the results indicated that restriction endonuclease analysis is a suitable typing method for C difficile, which may be very valuable in epidemiological studies where a highly discriminating typing method is needed. PMID- 3040812 TI - Use of hybridot assay to screen for BK and JC polyomaviruses in non immunosuppressed patients. AB - Urine samples from 50 patients attending a genitourinary outpatient clinic and from 13 renal allograft recipients were investigated for evidence of infection with human BK and JC polyomaviruses using cytology and a new DNA hybridot assay. Forty four per cent of samples from the renal allograft recipients were positive by cytology and 75% by DNA hybridisation, indicating that hybridot assay is more sensitive than cytological screening. BK and JC viral DNA was found in 20% of the patients attending the genitourinary clinic, showing infection with BK virus and JC virus in a group of patients with clinical conditions not normally associated with immunological deficiency-a finding that has not been reported before. PMID- 3040813 TI - Ultracytochemical distribution of ouabain-sensitive, K+-dependent, p nitrophenylphosphatase in the synaptic layers of goldfish retina. AB - Ouabain-sensitive, K+-dependent p-nitrophenylphosphatase (K+-pNPPase) activity, which represents the second dephosphorylation step of Na+,K+-ATPase, was localized histochemically at the light and electron microscopical levels in the goldfish retina. K+-pNPPase staining was most intense in the outer and inner plexiform layers and less intense over the photoreceptor inner segments. K+ pNPPase staining was observed on the membranes of horizontal cell dendrites and presynaptic membrane of all cone pedicles but only rarely over rod spherules. Bipolar cell dendrites in the outer plexiform layer were not stained for K+ pNPPase. In the inner plexiform layer (IPL), K+-pNPPase staining was observed at 90% of the bipolar cell ribbon synapses but only at 40% of amacrine cell synapses. The proportion of K+-pNPPase staining at amacrine cell synapses increased from 26 to 49% as one progressed from the outer to inner layers of the IPL, while staining at bipolar cell synapses showed no such trend. Only 16% of the amacrine synapses onto mixed, rod-cone (mb) bipolar cell synaptic terminals were positive for K+-pNPPase. We suggest that the differential distribution of K+ pNPPase staining at retinal synapses can be explained, in part, by the ionic conductances gated at the postsynaptic sites. In addition, the presence of K+ pNPPase on lateral horizontal cell dendrites in cone pedicles is consistent with the hypothesis that the sodium pump is involved in the release of GABA at feedback synapses from horizontal cells to cone photoreceptors. PMID- 3040814 TI - Dependence of cytochrome oxidase activity in the rat lateral geniculate nucleus on retinal innervation. AB - Patterns of cytochrome oxidase (CO) activity were examined histochemically in the dorsal lateral geniculate nucleus (LGNd) and retina of pigmented rats. CO staining was not uniform and was distributed in a pattern similar to that of retinal afferents. Portions of the LGNd receiving an exclusively crossed projection were moderately reactive whereas regions receiving an uncrossed or overlapping crossed and uncrossed projection were darkly reactive. The dependence of oxidative metabolic activity in the LGNd on retinal innervation was verified in animals with unilateral enucleation. In adults, chronic monocular enucleation led to a decrease in CO staining in portions of the LGNd deprived of retinal input; in animals enucleated at birth, normal patterns of CO reactivity failed to develop and both LGNds had a more uniform pattern of moderate CO staining. Most neurons in the ganglion cell layer of the retina were moderately reactive for CO. However, there were approximately 3,000 darkly reactive cells, most of which appear to be ganglion cells. The darkly reactive cells were more numerous in the peripheral temporal retina. The laminar pattern of CO staining in the retina was similar to that described previously for carnivores and primates. The most reactive laminae were the inner and outer plexiform layers and the photoreceptor inner segments. Within the inner plexiform layer, sublamina a was more darkly stained than sublamina b. These results suggest that the physiological properties of crossed and uncrossed visual pathways in rats are functionally dissimilar at the level of both the retina and the LGNd. PMID- 3040815 TI - The influence of food on the pharmacokinetics of itraconazole in patients with superficial fungal infection. AB - Itraconazole in a new, orally active triazole, which is related to ketoconazole and which is active against dermatophytes and Candida and Cryptococcus organisms. It is more potent than ketoconazole, is better absorbed with food, and does not suppress testosterone or cortisol synthesis. In previous studies, daily 200 mg doses of itraconazole for 5 days cleared 80% of cases of pityriasis versicolor, and 90% of dermatophyte infections were cleared by daily 100 mg doses administered for 1 month. Twenty patients with superficial dermatophyte, Candida albicans, and pityriasis versicolor infections were treated with doses of 50 or 100 mg per day; in the first group of 10 patients, the drug was administered before breakfast and the other 10 patients were given their dose with breakfast. Itraconazole concentration was determined by high-performance liquid chromatography 3 hours after administration of first and last doses. Our findings showed that although treatment with 50-mg doses led to poorer results and lower plasma levels than 100-mg doses, taking the drug with breakfast gave much better results than taking it before. PMID- 3040816 TI - Verrucous carcinoma of the skin associated with syringadenoma papilliferum: a case report. AB - We report the case of a 67-year-old man with a granular, cauliflower-pink lesion on the skin of the thigh. Histopathological study showed a typical verrucous carcinoma associated with a syringadenoma papilliferum. To our knowledge this association has not been previously reported. PMID- 3040817 TI - Regulation of hormone-stimulated cyclic AMP accumulation in intact human mononuclear leukocytes by blood plasma. AB - Several hormones, including catecholamines, histamine, and prostaglandin E1, regulate the function of human mononuclear leukocytes (MNL) by stimulating the accumulation of cAMP. Isoproterenol-stimulated cAMP accumulation in MNL isolated and washed at 4 degrees is five times greater than in cells prepared at ambient temperature. The current study was aimed at understanding this difference. cAMP accumulation in MNL prepared at ambient temperature could not be increased by chilling the cells for 4 hours. Warming MNL prepared at 4 degrees for 30 min, however, reduced later isoproterenol-, histamine-, and PGE1-stimulated cAMP accumulation by 65-85% without altering forskolin-stimulated cAMP accumulation and without altering cellular viability or ATP content. In broken cell preparations, there was no difference in either adenylate cyclase or phosphodiesterase activities, and no difference in the binding of isoproterenol to the beta-adrenergic receptors. The reduction in isoproterenol-stimulated cAMP accumulation in warmed intact cells was reversed when the MNL were incubated with autologous leukocyte-depleted blood or with plasma. These data suggest the presence of one or more factors in plasma that enhances hormone-stimulated adenylate cyclase activity in intact MNL. PMID- 3040818 TI - Insulin stimulation of cyclic AMP phosphodiesterase is independent from the G protein pathways involved in adenylate cyclase regulation. AB - The intact rat adipocyte was used to investigate the possibility of common intermediates in the insulin stimulation of cyclic AMP phosphodiesterase and the beta-adrenergic/adenosine regulation of adenylate cyclase. A five minute incubation of the isolated adipocytes with insulin produced a 50-100% increase in the phosphodiesterase activity found in the particulate fraction of homogenates. The insulin stimulation was not impaired by the presence of either agonist or antagonists of the inhibitory adenosine receptor which acts on adenylate cyclase. Phosphodiesterase activation by insulin was also observable above the level of stimulation produced by the beta-adrenergic agent isoproterenol and forskolin. The validity of the enzyme activity measurements was supported by measurements of the hormonal actions on cyclic AMP levels within the cells. Possible crossover between the adenylate cyclase and phosphodiesterase regulation systems at a post receptor site was investigated using adipocytes exposed to bacterial toxins specific for the modification of guanine nucleotide binding proteins. Both cholera toxin, which irreversibly activates Gs and pertussis toxin which inactivates Gi caused some stimulation of the phosphodiesterase activity and suppressed activation by isoproterenol, but neither toxin prevented the insulin stimulation of cyclic AMP phosphodiesterase. These results suggest, while common components may participate in the beta-adrenergic stimulation of both adenylate cyclase and phosphodiesterase, the mechanism of insulin activation of the phosphodiesterase does not involve the components of adenylate cyclase regulation. PMID- 3040819 TI - Calmodulin sensitive phosphodiesterase of porcine cerebral cortex: kinetic behavior, calmodulin activation, and stability. AB - The calmodulin sensitive phosphodiesterase of porcine cerebral cortex was characterized in terms of kinetic behavior, calmodulin activation, and stability. This enzyme displayed non-Michaelis-Menten kinetics in the presence or absence of calmodulin. The apparent affinity for cyclic GMP was higher than that for cyclic AMP but at saturating levels of substrate, this enzyme catalyzed the hydrolysis of cyclic AMP at a greater rate than it did cyclic GMP. The affinity of this enzyme for calmodulin was about 20-fold lower than usually reported. The apparent loss of phosphodiesterase activity after storage was found to be due to a strong association with container surfaces and could be prevented or reversed by the presence of 0.1% Triton X-100. PMID- 3040820 TI - Polyamine stimulation of protein phosphatase-2A from rat liver using a non protein phosphoester substrate. AB - The polyamines, spermine and spermidine, activate a high molecular weight form of phosphorylase a phosphatase isolated from rat liver. This broad specificity protein phosphatase (type 2A) was partially purified, using both protein and non protein phosphoester substrates. Spermine and spermidine activated isolated protein phosphatase-2A1 (apparent Mr 210,000) approximately 2-fold, when p nitrophenyl phosphate (PNPP) was used as substrate. Freeze-thawing, which activated the phosphatase activity against a variety of phosphoprotein substrates, also increased the extent of stimulation of PNPP phosphatase activity by spermine (8 to 9-fold with Ka of 93 microM) and spermidine (6 to 7-fold with Ka 280 microM). Kinetic analysis indicated that the activation of phosphatase by polyamines was accomplished by an increase in Vmax of the enzyme, by a mechanism independent of that achieved by other cations. The data indicate that polyamines, at physiological concentrations, can activate a form of protein phosphatase widely distributed in mammalian tissues, and thereby influence cellular protein phosphorylation. PMID- 3040821 TI - The effect of parathyroid hormone (1-34) on cyclic AMP level, ornithine decarboxylase activity, and glycosaminoglycan synthesis of chondrocytes from mandibular condylar cartilage, nasal septal cartilage, and spheno-occipital synchondrosis in culture. AB - Previously, we reported methods for isolating chondrocytes from the craniofacial complex and their culture in vitro. The response of these chondrocyte cultures to bovine parathyroid hormone (1-34) (PTH) has now been investigated. PTH stimulated glycosaminoglycan (GAG) synthesis, a characteristic of the cartilage phenotype in cultured chondrocytes isolated from mandibular condylar cartilage (MCC), nasal septal cartilage (NSC), and spheno-occipital synchondrosis (SOS). These stimulations of GAG synthesis by PTH were dose-dependent. PTH also increased accumulation of cyclic AMP (cAMP) and the activity of ornithine decarboxylase (ODC), a rate-limiting enzyme in polyamine biosynthesis. However, PTH did not stimulate DNA synthesis. The increases in the cAMP level, ODC activity, and GAG synthesis after addition of PTH (10(-7) mol/L) were greatest in MCC-chondrocytes and least in NSC-chondrocytes. The difference in the responses to PTH of these three types of chondrocytes may reflect differences of the characteristics of these cells in vivo. PMID- 3040822 TI - Thermal diffusivity of composite restorative materials. AB - The substantial increases in the filler volume fraction of the current generation of composite resins, and the incorporation of radiopacifying heavy elements in many of these fillers, constitute significant changes which may affect thermal transport properties. Thermal diffusivity has been determined for 21 of these composite materials recommended for anterior and posterior applications. For radiopaque hybrid and for microfine composites, there was, however, only a gradual trend to increased thermal diffusivity with increasing volume fraction of inorganic filler. The diffusivity values were not greatly in excess of the level observed for dentin. Nevertheless, a small group of materials, incorporating substantial amounts of quartz or silicon nitride filler particles, exhibited high rates of thermal diffusion, up to three times the level exhibited by dentin. PMID- 3040823 TI - Ontogeny and senescence of salivary immunity. AB - The objective of the present study was to evaluate the capacity for secretory immune responses throughout life. This was done by measuring, by single radial immunodiffusion, the concentrations of IgA and IgA1 subclass in saliva samples of subjects who ranged in age from two months to 91 years. The presence of salivary IgA antibodies to two nearly ubiquitous mucosal antigens, Streptococcus mutans glucosyltransferase (GTF) and killed poliovirus (Types 1, 2, and 3), was measured in an enzyme-linked immunosorbent assay in this population. Whole saliva from 2-5 month-old infants contained significantly less IgA than did parotid saliva of any adult group. Also, a significantly higher proportion of the total salivary IgA was IgA1 in infants' saliva than was found in parotid saliva of adults. Salivary IgA and IgA1 subclass levels in parotid saliva of young and old (70-91 years) adults did not differ. Salivary IgA antibody levels to GTF were negligible in most saliva samples of children less than five years old, while 40% of children older than one year had detectable levels of salivary antibody to poliovirus (PV). This differences between response to GTF and PV antigens may reflect differences in antigenic challenge. Parotid saliva of the oldest group (70-91 years) had narrowly distributed and uniformly low levels of IgA antibody to both antigens. Since their IgA immunoglobulin levels were the same as in younger adults, the low antibody levels in this oldest group may be associated with changes in the number or function of T or B lymphocytes or antigen-processing cells, and/or may result from diminished levels of challenge with these antigens. PMID- 3040824 TI - Antimicrobial factors in saliva: ontogeny and relation to oral health. AB - Antimicrobial agents (antibody and non-antibody) present in human saliva protect oral tissues by a variety of mechanisms, such as prevention of bacterial adhesion, agglutination of micro-organisms, and inhibition of multiplication and metabolism. However, studies in which the concentrations of various salivary antimicrobial agents have been correlated to the presence and severity of oral diseases--of dental caries, in particular--have produced controversial data, and it seems evident, also on the basis of the present study, that no single salivary antimicrobial factor (except flow rate) affects oral health to a significant degree. In the present study, we report the levels of some selected salivary antimicrobial agents in predentate and dentate human infants, with a comparison to the levels found in young adults' saliva. Salivary lysozyme, peroxidase, and hypothiocyanite concentrations were already at the adult level at the time when the primary teeth erupt, whereas immunoglobulin (IgA, IgG, and IgM), lactoferrin, myeloperoxidase, and thiocyanate concentrations were significantly lower in children than in adults. Dentate children had more IgG, thiocyanate, and protein in whole saliva than did predentate children. PMID- 3040825 TI - Effects of variations in pH and hypothiocyanite concentrations on S. mutans glucose metabolism. AB - Hypothiocyanous acid (HOSCN) and hypothiocyanite (OSCN-) were generated by the antibody-independent salivary peroxidase (SP) system. The metabolism of Streptococcus mutans NCTC 10449 was examined by uniformly labeled glucose incorporation studies. We found that the SP-system causes a pH-dependent inhibition of 14C-labeled glucose uptake, and that the effects of HOSCN/OSCN- are bacteriostatic. The results also showed that, at low pH, bacteria required more time to recover fully from HOSCN/OSCN- inhibition. When control experiments were performed in the absence of HOSCN/OSCN-, but the pH was varied, we found a positive correlation between pH and the rate of 14C-glucose incorporation. The results also showed that pH did not affect the maximum incorporation of 14C glucose, demonstrating that S. mutans can adapt to pH changes in the environment. Based on the data obtained, we postulate that the antibody-independent SP system plays an important role in the regulation of the metabolism of oral streptococci. PMID- 3040826 TI - Interference of IgA protease with the effect of secretory IgA on adherence of oral streptococci to saliva-coated hydroxyapatite. AB - It has previously been shown that secretory immunoglobulin A (S-IgA) influences the sorption of oral streptococci to hydroxyapatite as well as to cell surfaces. The present experiments demonstrate that bacterial IgA proteases, which cleave S IgA in the hinge region, are capable of interfering with this mechanism. This result was obtained with an IgA1 specific protease from Haemophilus influenzae and with a protease from Clostridium ramosum that cleaves IgA1 as well as IgA2 of A2m(1) allotype. The modulation of S-IgA-mediated effects by IgA proteases were studied by means of an in vitro method which permits quantitative determination of the sorption of radiolabeled oral bacteria to hydroxyapatite beads. Other authors have suggested that IgA protease-mediated effects may be explained by a strongly reduced antigen-binding capacity of released Fab alpha fragments. Here we present evidence that streptococci, after exposure to specific S-IgA and IgA protease, are coated with Fab alpha fragments. PMID- 3040827 TI - Regulatory aspects of N-linked glycoproteins. AB - Activation of beta-adrenoreceptors in rat parotid acinar cells leads to copious exocrine protein secretion. Additionally, beta-adrenergic stimulation dramatically increases specific secretory protein synthesis and enhances N-linked glycosylation of secretory glycoproteins. Recently, efforts have been directed toward understanding the mechanisms underlying these biosynthetic events. We have been particularly interested in the receptor-mediated regulation of glycosylation. In this report, we evaluate available mechanistic information from the rat parotid gland and present initial data examining the ability of various regulatory agents to modulate N-linked glycosylation in enzymatically-dispersed cell aggregates from surgical specimens of human parotid glands. We conclude that glycosylation of human parotid N-linked glycoproteins may be regulated by extracellular signaling similar to that operative in the rat parotid gland. PMID- 3040828 TI - The effects of increased intraluminal pressure on rat submandibular gland morphology. PMID- 3040829 TI - Role of protein phosphorylation in rat salivary gland exocytosis. PMID- 3040830 TI - Effect of cholesterol feeding on membrane fluidity, (Na+ + K+)ATPase, adenylate cyclase, [3H]-ouabain-, and [3H]-dihydroalprenolol-binding in rat submandibular salivary glands. PMID- 3040831 TI - Initial characterization of a neutral metalloproteinase, active on native 3/4 collagen fragments, synthesized by ROS 17/2.8 osteoblastic cells, periodontal fibroblasts, and identified in gingival crevicular fluid. AB - Analysis of collagenolytic activity in gingival crevicular fluid (GCF) has revealed the presence of an enzyme capable of fragmenting native 3/4- and 1/4 collagen cleavage products generated by collagenase. An enzyme with similar activity was also identified in media conditioned by fibroblasts from rat periodontal ligament and gingiva, and by rat osteoblastic cells (ROS 17/2.8, 17/2A, 17/2B). In culture, the enzyme was secreted in a latent form that could be activated by organomercurials. For further characterization of this novel enzyme (MMP-V), the osteoblast proteinase was partially purified. ROS 17/2.8 conditioned medium was harvested daily and the 25%-60% sat. ammonium sulfate fraction chromatographed on an AcA 54 gel filtration column. Latent forms of MMP-V (apparent Mr approximately 54 k) and collagenase (Mr approximately 54 k) were resolved from gelatinase (Mr approximately 76 k) and two collagenase inhibitors (Mr approximately 62 k, approximately 36 k). Activated MMP-V degraded native 3/4 collagen fragments from collagen types I and II in a step-wise manner and was active on denatured collagen. MMP-V showed a divalent cation requirement, was active at neutral pH, and was inhibited by collagenase inhibitor and fetal bovine serum, but not by serine, thiol, or carboxyl proteinase inhibitors. These properties indicate that MMP-V is a member of the matrix-degrading, neutral metalloproteinase family of enzymes which include collagenase, gelatinase, stromelysin, and telopeptidase. The enzyme may function in the degradation of collagen fibrils by cleaving proteinase-resistant 3/4-collagen fragments that are stabilized by association with neighboring collagen molecules. PMID- 3040832 TI - A non-antibacterial chemically-modified tetracycline inhibits mammalian collagenase activity. AB - Tetracyclines (including the semi-synthetic analogues, minocycline and doxycycline) are considered useful adjuncts in periodontal therapy because they suppress Gram-negative periodontopathogens. Recently, these antibiotics were found to inhibit mammalian collagenase activity, a property which may also be of therapeutic value. It has been suggested that the anti-collagenase properties of the tetracyclines are independent of their antibiotic efficacy. To advance this hypothesis further, we chemically converted tetracycline hydrochloride to its non antimicrobial analogue, de-dimethylaminotetracycline. This chemically-modified tetracycline (CMT), although no longer an effective antibiotic, was found to inhibit the in vitro activity of collagenase from partially purified extracts of human rheumatoid synovial tissue and rachitic rat epiphysis. In a preliminary in vivo study, pathologically-excessive collagenase in skin and gingiva was induced by rendering adult male rats diabetic, and the oral administration of CMT to these rats significantly reduced the excessive collagenase activity in both tissues. Moreover, CMT administration did not affect the severe hyperglycemia in these rats but did prevent, at least in part, the diabetes-induced loss of body weight, skin weight, and skin collagen mass; these effects suggest a lack of toxicity in this animal model. A proposed clinical advantage of CMT over conventional tetracyclines, in the treatment of diseases characterized by excessive collagenolytic activity, is the lack of development of antibiotic resistant micro-organisms during prolonged use. However, the consideration of clinical trials to support this hypothesis must await further laboratory and extensive toxicity tests. PMID- 3040833 TI - Effect of acid type on kinetics and mechanism of dental enamel demineralization. AB - The influence of acid type (pKa effects) of weak organic acid buffers on dissolution kinetics of dental enamel was critically examined for rigorous testing of the behavioral validity of the physical model of Patel et al. (1987). Quantitative evaluation of this model indicated that monitoring initial dissolution rates was a viable approach to critical testing of the model. Initial dissolution rates were determined in 0.1 mol/L acetate (pKa = 4.77), benzoate (pKa = 4.20), and salicylate (pKa = 2.98) buffers (pH = 4.50, mu = 0.50), with ground bovine enamel blocks of known surface area mounted in a rotating disk apparatus. The Levich theory was used to study dependence of dissolution rates on stirring rates in these buffers. The experimental data were analyzed by the physical model which includes pKa effects, complexation of the buffer anion with the other ions, surface kinetics, simultaneous diffusion and equilibrium of all species in enamel pores, diffusion layer thickness, and bulk solution composition. The KIAP (formula: see text) governing the dissolution reaction and the surface resistance factor were deduced from the model. Dissolution kinetics was also followed in these buffers in the presence of calcium or phosphate common ions. In effect, by conducting both the stirring rate studies and common ion experiments, we derived the driving force function independently by these two techniques. The results obtained in this study were consistent with the model, indicating that pKa effects on the dissolution of dental enamel can be accounted for quantitatively by the model, and it was found that weak acids do not influence either the apparent solubility or the surface reaction process of bovine dental enamel. PMID- 3040834 TI - Sodium absorption. PMID- 3040836 TI - Intravenous desensitization to beta-lactam antibiotics. AB - Patients allergic to penicillin (PCN) often require treatment with beta-lactam antibiotics for life-threatening bacterial infections. In this article, we review our experience with rapid intravenous desensitization for patients who gave a history of PCN allergy and who had hypersensitivity demonstrated by skin tests. Skin testing was performed with both prick and intradermal techniques and with the recommended antibiotic as well as PCN G, penicilloyl polylysine, and a minor determinant mixture. Patients were transferred to the intensive care unit, and desensitization was performed with a buret technique that required minimal preparation and was easily applied to any antibiotic. Fifteen desensitizations in 12 patients were associated with no immediate reactions. One patient developed a delayed reaction consisting of a pruritic rash and angioedema. A second patient developed a more serious delayed serum sickness-like illness with fever, rash, eosinophilia, abnormal liver function tests, and urinary abnormalities. These reactions did not necessitate stopping the antibiotic, although the latter patient required corticosteroids to suppress his symptoms. Rapid intravenous desensitization is a rapid, safe, and effective technique for patients demonstrating hypersensitivity to beta-lactam antibiotics who require therapy with these medications. PMID- 3040835 TI - Prednisone inhibits late asthmatic reactions and airway inflammation induced by toluene diisocyanate in sensitized subjects. AB - To determine the importance of airway inflammation for late asthmatic reactions induced by toluene diisocyanate (TDI), we investigated whether prednisone prevented them [corrected] by modifying the associated airway inflammatory reaction. We measured FEV1 before and at regular intervals after exposure to TDI and performed bronchoalveolar lavage at 8 hours after TDI in two groups of subjects with previously documented late asthmatic reactions, in one group, after no treatment, and in the other group, after treatment with prednisone (50 mg/day for 4 days). After no treatment, each subject developed a late asthmatic reaction, an increase in airway responsiveness, polymorphonuclear leukocytosis, and increased albumin in bronchoalveolar lavage. By contrast, after treatment with prednisone, no subject developed a late asthmatic reaction or an increase in airway responsiveness, and the number of leukocytes and the concentration of albumin were normal in bronchoalveolar lavage. These results suggest that late asthmatic reactions induced by TDI may be caused by airway inflammation and that prednisone may block them [corrected] by inhibiting the inflammatory reaction of the airway induced by TDI in sensitized subjects. PMID- 3040837 TI - Regulation of the beta-receptor-adenylate cyclase system in lymphocytes of allergic patients with asthma: possible role for protein kinase C in allergen induced nonspecific refractoriness of adenylate cyclase. AB - Allergen challenge of allergic patients with asthma caused various changes in the beta-receptor-adenylate cyclase system of lymphocyte membranes from these patients. These changes included uncoupling and down regulation of beta adrenergic receptors and nonspecific refractoriness of adenylate cyclase, as demonstrated by reduced responses to isoproterenol (beta 2), histamine (H2), 5' guanylylimidodiphosphate, and sodium fluoride. Since these changes could be due to desensitization by enhanced plasma levels of catecholamines and/or histamine during the allergic response, we explored the effects of these agonists on the beta-receptor-adenylate cyclase system in vitro with normal lymphocytes. In addition, we assessed the effect of the tumor-promoting phorbol ester, phorbol 12 myristate 13-acetate (PMA), on this system, since phorbol esters have been demonstrated to modulate several receptor systems, presumably via activation of protein kinase C. That both the agonists and PMA may cause refractoriness of lymphocyte adenylate cyclase was demonstrated, but, however, by apparently different mechanisms. The agonists isoproterenol and histamine induced only a specific desensitization of the homologous responses, whereas PMA-induced refractoriness was nonspecific in nature. Radioligand-binding studies demonstrated that both uncoupling and down regulation contributed to the isoproterenol-induced beta-adrenergic hyporesponsiveness, whereas beta-adrenergic receptor uncoupling but not beta-adrenergic receptor down regulation was involved in PMA-induced desensitization. Histamine had no effect on the beta-adrenergic system at all. The data suggest that the agonist-induced changes in the adenylate cyclase system are specifically located at the receptors, whereas PMA-induced refractoriness can be explained by alterations distal to the receptors, presumably at the stimulatory guanine nucleotide regulatory protein. Thus, enhanced levels of catecholamines or histamine could be involved in the development of receptor-specific changes in the lymphocyte adenylate cyclase system of allergic patients with asthma. However, they are unlikely to cause the nonspecific changes distal to the receptors. The latter changes could be induced by physiologic activation of protein kinase C during the allergic response by a still unknown stimulus, possibly via the receptor-mediated turnover of phosphatidylinositol 4,5-diphosphate. PMID- 3040838 TI - Dietary fiber: comments on interpreting recent research. PMID- 3040839 TI - Dietary fiber: classification, chemical analyses, and food sources. AB - Dietary fiber's role in the prevention and treatment of constipation has long been known, but now fiber is touted as a cure for many of the ills in Western countries. Although some data exist to relate dietary fiber intake to certain diseases, lack of agreement on what dietary fiber is and how it should be measured make interpreting the data difficult. Further, not all dietary fiber is created equal. Water-soluble fibers, such as pectin and gums, have little effect on stool weight and hence are not appropriate treatment for patients with constipation. Water-insoluble fibers, such as cellulose and hemicellulose, are most effective in aiding laxation but may also limit absorption of minerals and possibly vitamins. Wheat bran is a good source of hemicellulose; vegetables supply cellulose to the diet. Most agencies are recommending a doubling or tripling of dietary fiber intake. Typical recommendations are set at 25 to 50 grams of dietary fiber daily. Different analytical methods for dietary fiber yield conflicting fiber values, and dietary fiber values do not exist for many foods, making fiber recommendations controversial and difficult to achieve. Fiber in the diet should ideally be increased by the consumption of unrefined breads and cereals and more fruits and vegetables. Vegetarians routinely consume 40 to 50 gm dietary fiber daily without ill effect. Fiber supplements may be appropriate for some patients, but the composition of the fiber should be known and be appropriate for the disease being treated. Before fiber supplements are marketed, clinical trials should be conducted to support the use of the supplements in the prevention and treatment of disease.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3040840 TI - The role of dietary fiber in gastrointestinal disease. AB - The most consistent benefit of consumption of adequate dietary fiber is regular laxation; this effect alone justifies inclusion of fiber in the diet, in view of the enormous expenditure on drugs for digestive diseases. Dietary fiber has proved effective in decreasing symptoms of diverticular disease, Crohn's disease, and hemorrhoids in a limited number of small clinical studies. Fiber may also reduce the incidence of gallstone formation. Fiber is currently being touted as protection against colon cancer. However, the epidemiological and experimental data do not provide convincing evidence that fiber alone is a major determinant of risk for colon cancer. Furthermore, the data from international comparisons indicating that fiber is protective against colon cancer can be used in a similar simplistic manner to suggest that fiber may be a risk factor for stomach cancer. This should not dissuade individuals from obtaining adequate fiber from a wide variety of foods but should caution them against consumption of excessive amounts of fiber from a single source or from dietary supplements. PMID- 3040841 TI - Dietary fiber in the reduction of colon cancer risk. AB - The evidence for an inverse association between a diet of foods high in fiber and colon cancer risk is reviewed in the context of the epidemiological criteria for causality. Five criteria are assessed: consistency of the association, strength of the association, specificity of the hypothesis, temporal relationship of the association, and coherence of the association. Forty epidemiological studies, described in 55 original reports, are analyzed in terms of an association between fiber intake and colon cancer. This evaluation clearly suggests a relationship between colon cancer and diet low in fiber. The epidemiological studies focus on dietary patterns in which fiber usually occurs as a complex mixture with other foods. At present, information on the chemistry and function of various types of fiber as well as the mechanisms of cancer inhibition still is quite limited. As dietary fiber may interact with or be linked to other dietary factors, the impact of total diet and dietary interactions should be considered in studies of colon cancer risk and in dietary counseling. PMID- 3040842 TI - Dietary fiber and diabetes: a comprehensive review and practical application. AB - Diabetes takes an enormous human and monetary toll each year. Current treatment too often revolves around insulin and drug therapy, neglecting diet and exercise. A comprehensive review was undertaken to assess and summarize the effects of dietary fiber on diabetes. Fiber supplement studies with guar, wheat bran, and apple fiber produced mixed results; some studies reported lowered fasting glucose and cholesterol values and less glycosuria. However, many available fiber supplements cause side effects. High-carbohydrate, high-fiber diets providing 55% to 60% of energy as carbohydrates, 15% to 20% as protein, and 20% to 25% as fat and including 50 gm or more fiber daily hold the most potential for long-term use. These diets reduce insulin requirements, improve glycemic control, lower fasting serum cholesterol and triglyceride values, and promote weight loss. Studies show good long-term adherence with these diets. Dietitians assume primary responsibility for educating individuals on the benefits and use of high carbohydrate, high-fiber diets. Diets must be individualized, with special modifications for obesity, hyperlipidemia, or physiological states such as pregnancy and lactation. Widespread use of high-fiber diets will ultimately improve metabolic control and decrease health care costs for thousands of diabetic and non-diabetic individuals. PMID- 3040844 TI - The effect of lead on chemical- and viral-induced tumor production in mice. AB - Female Swiss mice were exposed to lead in the drinking water at concentrations ranging from 0 to 1000 ppm for 105 or 280 day periods of time. The effect of lead on urethan-induced pulmonary adenoma formation was evaluated in the 105 day study. Urethan-induced sleeping times observed following ip injection of urethan (1.5 mg/g) after 3 weeks of lead exposure were not altered by lead indicating that lead did not affect the rate of urethan elimination. Pulmonary adenoma formation was evaluated 84 days later. Lead exposure did not affect the number of tumors produced, nor did it alter the mean tumor diameter in the lead treatment groups. This suggests that the immunosuppressive activity of lead does not enhance urethan-induced adenoma formation. In the 280 day study, the incidence of spontaneous murine lymphocytic leukemia was evaluated. Leukemia was observed in all treatment groups. Mortality was greater in the lead-exposed mice. Mice exposed to 50 or 1000 ppm lead had 41.6% and 58.3% more deaths associated with the virus. The median survival time was also reduced in the lead-exposed mice. It appears that the immunosuppressive effects of lead allow for increased expression of the murine lymphocytic leukemia virus. PMID- 3040843 TI - Interrelationship between hemolysis and lipid peroxidation of human erythrocytes induced by silicic acid and silicate dusts. AB - Silicic acid and silicate dusts (slate dust and chrysotile asbestos) cause hemolysis of erythrocytes in vitro. The peroxidation of polyunsaturated fatty acids (PUFA) of erythrocyte membrane lipids is also enhanced by incubating the erythrocytes with silicic acid and silicate dusts in in vitro. Hemolysis of erythrocytes elicited by silicic acid and silicate dusts is inhibited significantly by polyvinyl-pyrrolidone and dipalmitoyl lecithin (DPL). These agents, however, have no effect on silicic acid and silicate dust induced peroxidation of erythrocyte membrane lipids. On the other hand, peroxidation of erythrocyte membrane lipids, induced by silicic acid and silicate dusts, is inhibited almost completely by adding superoxide dismutase and catalase to the incubation system, whilst the hemolysis of erythrocytes induced by silicic acid and silicate dusts is unaffected by these agents. Similarly the lysis of erythrocytes, induced by silicic acid and silicate dusts, proceeds at a much faster rate than silicic acid and silicate dust induced lipid peroxidation. These results indicate that silicic acid and silicate dust induced hemolysis and lipid peroxidation represent two independent processes. PMID- 3040845 TI - Effects of in vivo administration of detergents on the hepatic microsomal cytochrome P-450 system in rat. AB - The influence of in vivo administration of detergents on the hepatic microsomal cytochrome P-450 system was studied in rat. Male Wistar rats were administered detergents, Emulgen 913 (50 mg or 100 mg kg-1 of body weight (B.W.], or sodium dodecylsulfate (SDS, 25 mg or 50 mg kg-1 of B.W.) intraperitoneally once a day for 3 days. Cytochrome P-450 content in liver microsomes was significantly decreased to 85% and 73% of control by the administration of 50 mg or 100 mg Emulgen, respectively, but the microsomal protein concentration was not changed by these administrations. The content of cytochrome P-450 was also reduced to 76% and 70% of control by the administration of 25 mg or 50 mg SDS/kg of B.W., respectively. The total hydroxylation activity (the sum of omega- and (omega-1) hydroxylase activity) of laurate almost paralleled the decrease in cytochrome P 450 in detergent-treated rats. However, the omega/omega-1-hydroxylation ratio was not changed. These results suggest that the administration of these detergents lowered the level of cytochrome P-450 species catalyzing omega- and (omega-1) hydroxylation of laurate to a similar extent. On the other hand, aminopyrine N- and p-nitroanisole O-demethylation activities in Emulgen 913-treated rats was decreased while those in SDS-treated rats did not change, though the content of cytochrome P-450 was decreased by both administrations. Thus, it was demonstrated that the livers of rats responded to exogenous detergents in different manners. PMID- 3040846 TI - The combined effect of Pb2+ and Mn2+ on monoamine uptake and Na+, K+-ATPase in striatal synaptosomes. AB - Rat striatal synaptosomes (P2-fraction) were subjected to lipoperoxidation by the addition of 120 microM Fe2+ and 200 microM ascorbic acid. This preparation (pretreated synaptosomes) was used to investigate the interaction of Pb2+ and Mn2+ on the uptake of tritiated catecholamines, Na+, K+-ATPase activity and malondialdehyde (MDA) formation in order to understand the mechanism of enhanced neurotoxicity by concurrent exposure to these metals. The combination of Pb2+ and Mn2+ (25 microM + 100 microM, respectively) produced a significant increase in the uptake of 3H-Dopamine only in the untreated synaptosomes. No significant effect was noted on the uptake of 3H-Norepinephrine in either pretreated or untreated synaptosomes. However, the combination of Pb2+ and Mn2+ produced a pronounced decrease in the activity of Na+, K+-ATPase, but the magnitude of the change was the sum of the individual metal effects. Metal interaction did not produce any significant change in the formation of MDA compared to the control (without addition of metals). These results indicate that Pb2+ and Mn2+ interaction may produce inhibition in the activity of transport ATPase in both the preparation of synaptosomes, with more pronounced effect of synaptosomes subjected to lipoperoxidation and these changes may be responsible for the disruption in the physiology of nerve impulse transmission. PMID- 3040847 TI - Enhancement of central transmission to sympathetic preganglionic neurons by phosphodiesterase inhibitors and its prevention by clonidine. AB - The effects of 3 phosphodiesterase inhibitors, aminophylline, isobutylmethylxanthine (IBMX), and RO 20-1724, were tested on descending intraspinal and spinal reflex transmission to sympathetic preganglionic neurons in unanesthetized spinal cats. Sympathetic discharges, recorded from upper thoracic preganglionic white rami, were evoked by stimulation (0.1 Hz) of descending excitatory pathways in the cervical dorsolateral funiculus (intraspinal) or of adjacent intercostal nerves (spinal reflex). Each phosphodiesterase rapidly and markedly enhanced transmission through intraspinal pathways but only slowly and modestly enhanced transmission through spinal reflex pathways. Pretreatment with a methyltyrosine-reserpine combination, chlorpromazine, or prazosin markedly restricted the enhancement of intraspinal transmission by IBMX to levels typically produced on spinal reflex pathways. Clonidine markedly depressed transmission through both pathways and prevented enhancement by the phosphodiesterase inhibitors. Yohimbine or tolazoline antagonized the depressant effects of clonidine and restored the ability of the phosphodiesterase inhibitors to enhance transmission. Somatic spinal reflexes were not affected by the phosphodiesterase inhibitors. The results suggest that descending norepinephrine pathways to sympathetic preganglionic neurons activate adenylate cyclase to generate cyclic AMP which increases neuronal excitability, possibly by phosphorylating membrane proteins. Clonidine appears to depress neuronal excitability by inhibiting adenylate cyclase through activation of alpha 2-adrenergic receptors. PMID- 3040848 TI - Linear Percoll gradient centrifugation of rat anterior pituitary cells. A simple method for prolactin cell enrichment. AB - Prolactin (PRL) cells were purified from nulliparous normal female adult Wistar rat pituitary cell suspensions by linear Percoll density gradient centrifugation, a procedure yielding single cells. Lactotrophs were found in two different layers, the first containing 70% PRL cells in the density range 1.055 to 1.065 g/ml, the second with 28% PRL cells in the range 1.070 to 1.080 g/ml. Both cell fractions contained more than 90% viable cells with an intact ultrastructure. The physiological integrity of the 70% enriched PRL cells was assessed by their basal PRL secretion, their secretory response to TRH and dopamine, and their cAMP production in a basal situation and after incubation with dopamine. PMID- 3040849 TI - Ultradian plasma corticotropin and cortisol rhythms: time-series analyses. AB - 24-h plasma patterns of ACTH and cortisol were established in 6 subjects who had standard meals and in 4 subjects under continuous enteral nutrition. Temporal relationships between both hormones were analyzed. The individual assay data were subjected to time-series analyses to identify the periods of the oscillations in the plasma levels. The spontaneously occurring cortisol peaks were preceded by increases in ACTH levels, but z-score transformations clearly revealed that ACTH and cortisol were not quantitatively linked throughout the day. Individual subjects' power spectra gave evidence of a predominant periodicity in the oscillations of both hormones. These periodicities varied between individuals. They were 55-140 min for ACTH and 95-180 min for cortisol, indicating that, on occasion, a single cortisol peak may be initiated by two ACTH peaks. Cross spectral analysis of the individual data gave coherence spectra with a large peak which accounted for a substantial concordance of their period length, and cross variance spectra showed a consistent phase relationship between both hormones. These time series analyses applied to the data further support the concept that ACTH is the stimulating factor of cortisol release under basic conditions. PMID- 3040852 TI - Effect of aging on hepatic carbohydrate metabolism in septic rats. AB - Aged individuals have diminished resistance to severe sepsis and septic shock. Past work with animals suggested that an important determinant of survival was the ability of the liver to supply glucose. In this study, young adult (3 to 4 months) and old (24 months) Fischer 344 rats were fasted and subjected to cecal incisions producing a rapidly lethal peritonitis. We then determined gluconeogenic intermediates in the liver. In the old rats with peritonitis, hexosemonophosphates (HMP) increased 50% relative to control liver, whereas in the young animals with peritonitis, the substrate decreased 50%. The accumulation of HMP in the old rat liver cells indicates a failure to dephosphorylate glucose 6-phosphate (G6P). This increase in HMP is associated with a decline in hepatic glucose-6-phosphatase (G6Pase), the final enzyme in the gluconeogenic pathway, and is reflected in a significant reduction in serum glucose in old Fischer 344 rats when compared to young Fischer rats. PMID- 3040850 TI - Treatment of pituitary macroadenomas secreting PRL, HGH or ACTH with long-acting bromocriptine. AB - Long-acting im bromocriptine was administered to 7 patients with pituitary macroadenomas (4 acromegalics, 1 Nelson's syndrome and 2 prolactinomas), with good tolerance except during the first 24 h. During a 42-day period hormonal, CT scan and visual field variations were followed. In acromegalics HGH decrease was not evident, except in some isolated sample. In Nelson's syndrome ACTH showed a 94% fall on day 14, even though a spontaneous oscillation cannot be ruled out, and recovery took place from day 21 on. PRL remained undetectable in both. In prolactinomas, PRL suffered a great decrease (91.8% and 96.3% on days 21 and 28 respectively) and remained well below its initial values up to the end of the study, in spite of partial recovery. In these 2 patients CT-scan evidenced shrinkage of tumor mass, which was not observed in the remaining 5 cases. Visual fields did not improve in the 2 cases initially affected (Nelson's syndrome and 1 prolactinoma). Long-acting bromocriptine seems to have the same therapeutic uses of the oral form with the possible advantage of a better tolerance of full initial doses. PMID- 3040851 TI - Corticotropin releasing hormone stimulation test: diagnostic aspects in Cushing's syndrome. AB - The effect of exogenous ovine Corticotropin-Releasing Hormone (oCRH) on plasma ACTH and cortisol levels was investigated in 10 normal volunteers and in 37 patients with Cushing's syndrome (26 with pituitary-dependent disease, 5 with an adrenal adenoma, 2 with an adrenal carcinoma and 4 with bilateral nodular hyperplasia). In all normal subjects and in patients with Cushing's disease, oCRH 100 micrograms as a bolus produced an increase in both plasma ACTH and cortisol. The peak of ACTH occurred after 15-30 min, while plasma cortisol showed highest levels between 30 and 60 min after oCRH administration. The hormonal response in Cushing's disease showed great variability with a clear hyperresponsiveness at least in 6 out of 26 patients with Cushing's disease. A slight and delayed response occurred in 3 cases of bilateral nodular adrenal hyperplasia, while a fourth case showed hyperresponsiveness similar to that found in pituitary dependent Cushing's disease. No response was observed in patients with an adrenal tumor. Eleven patients with Cushing's disease were tested before and 1 month after pituitary microadenomectomy. After surgery basal cortisol levels were reduced in 10 and became unresponsive or less responsive to oCRH. ACTH patterns were variable with a normal response only in few cases. Although this test seems of limited value in the diagnosis of hypercortisolism, it is a useful tool to differentiate some types of Cushing's syndrome (adrenal tumor from pituitary dependent Cushing's disease). Variable patterns of response in cases with bilateral nodular adrenal hyperplasia limit the usefulness of this test in recognizing this rare form of hypercortisolism. PMID- 3040853 TI - [Adenoid cystic carcinoma or cylindroma of the cervix uteri. Review of the literature apropos of a personal case]. AB - The authors report on a case of cylindroma of the cervix combined with a carcinoma in situ. Reviewing the literature which helped them, they studied the anatomo-pathological characteristics as well as the histogenesis and the therapy for cylindromas, which are more often found in other parts of the body. In particular, they are found in the salivary glands. There the origin is still uncertain and the evolution from the same stage is worse than in epidermoid carcinoma of the cervix. PMID- 3040854 TI - Antigen-induced proliferation and immunoglobulin A secretion by a human-human hybridoma. AB - The capacity of membrane immunoglobulin A (IgA)-bearing B cells to respond to specific antigen in the absence of T cell influences has not been defined. A human-human hybridoma, constructed from an Epstein-Barr virus transformed tonsil B cell that secreted IgA anti-phosphorycholine (PC) and a human plasmacytoma cell, was utilized to examine this issue. The cloned hybridoma expressed membrane IgA and secreted IgA specific for PC. Stimulation of the hybridoma cells with PC conjugated to Sepharose beads (PC-Sepharose) but not glycine-conjugated Sepharose resulted in an increase in DNA synthesis. Affinity purified goat anti-human IgA bound to Sepharose also augmented DNA synthesis. Soluble PC did not increase DNA synthesis and inhibited the increase in DNA synthesis resulting from PC Sepharose. IgA secretion was augmented in response to PC-Sepharose, as demonstrated by an increase in the number of Ig-secreting cells detected by a reverse hemolytic plaque assay and by quantitation of the IgA secreted per cell by enzyme-linked immunosorbent assay. Mitogen-stimulated T cell supernatants increased IgA secretion of the hybridoma cells but did not cause synergistic stimulation of the cells in the presence of PC-Sepharose. These data indicate that Sepharose-bound antigen was sufficient to induce proliferation and augment IgA secretion by this membrane IgA anti-PC-bearing hybridoma. The results suggest that cross-linking of membrane IgA by specific antigen may be a sufficient stimulus for proliferation and differentiation of B cells at this stage of maturation. PMID- 3040855 TI - A urine inhibitor of interleukin 1 activity affects both interleukin 1 alpha and 1 beta but not tumor necrosis factor alpha. AB - Urine from monocytic leukemia and other febrile patients contains an inhibitor of interleukin 1 (IL-1), as measured by prostaglandin E2 and collagenase production by human fibroblasts and synovial cells. With the use of recombinant IL-1, the IL 1 inhibitor was partially purified by using ammonium sulfate precipitation, anion exchange, and gel filtration chromatographies. IL-1 inhibitory activity elutes with an 18,000 to 25,000 apparent molecular size. The same fractions also inhibit IL-1 assayed by the proliferation of murine thymocytes and human fibroblasts. Both forms of human recombinant IL-1, IL-1 alpha and IL-1 beta, which show only 26% homology, but nevertheless bind to the same receptor, are affected by this natural inhibitor to the same extent. In contrast, human recombinant tumor necrosis factor, which shares some of the biologic activities of IL-1, is not inhibited by the urinary IL-1 inhibitor. This study shows that the various biologic activities of both forms of human recombinant IL-1 are inhibited by a partially purified natural urine-derived factor. PMID- 3040856 TI - Direct activation of human resting T cells by IL 2: the role of an IL 2 receptor distinct from the Tac protein. AB - High concentrations of interleukin 2 (IL 2) were shown to produce a delayed but pronounced proliferation of purified resting T cells in the apparent absence of other activation signals. Because these stimulatory effects of IL 2 occurred in the absence of detectable Tac+ cells, the possibility that IL 2 might be initially interacting with an IL 2 binding protein distinct from the Tac protein was studied. Chemical cross-linking studies with 125I-IL 2 revealed the presence of an IL 2 binding protein distinct from the Tac protein on the surface of these unstimulated T cells. This second IL 2 receptor has an estimated molecular size of 70,000 daltons, lacks reactivity with the anti-Tac antibody, and appears to be identical to the p70 protein recently proposed as a component of the high affinity IL 2 receptor. Scatchard analysis of IL 2 binding assays performed with the unactivated T cells revealed approximately 600 to 700 p70 sites per cell and an apparent Kd of 340 pM. These data indicate that the p70 protein present on resting T cells binds IL 2 with an intermediate affinity compared with the previously recognized high and low affinity forms of the receptor and may account for the high concentration of IL 2 needed to induce resting T cell proliferation. To investigate the early biologic consequences of IL 2 binding to the p70 protein, potential changes in the expression of genes involved in T cell activation were examined. Northern blotting revealed the rapid induction of c myc, c-myb, and Tac mRNA after stimulation of resting T cells with a high concentration of IL 2. The anti-Tac antibody did not inhibit IL 2 induced expression of these genes, suggesting that the p70 protein rather than the Tac antigen or the high affinity IL 2 receptor complex mediated this signal. However, in contrast to these early activation events, the anti-Tac antibody significantly inhibited IL 2 induced T cell proliferation. This finding implicates the high affinity form of the IL 2 receptor in the proliferative response of the IL 2 activated T cells. Thus these data support a two step model for the induction of resting T cell proliferation by high doses of IL 2 involving the initial generation of an activation or "competence" signal through the p70 protein and a subsequent proliferation or "progression" signal through the high affinity form of the receptor. PMID- 3040857 TI - Interleukin 2 activates a receptor-associated protein kinase. AB - The interleukin 2 (IL 2) receptor complex has been shown to consist of at least two IL 2 binding molecules, one of 55 to 57 kd (gp57Tac) and one of 75 to 78 kd apparent m.w. The data presented here indicate that a protein of m.w. 78,000 (pp78) co-immunoprecipitates with gp57Tac when a monoclonal antibody against gp57Tac is used. The 78 kd molecule is phosphorylated in vitro within the immune complex only in the presence of exogenously added IL 2, whereas the 57 kd molecule is phosphorylated equally in the presence or absence of IL 2. Phosphorylation in vitro of pp78 was demonstrated in extracts of human peripheral blood T cells (PBL-T) and the human T cell line Jurkat, but not in extracts of the human macrophage line U937 or the murine T cell line 2.8.2. Metabolic phosphorylation in intact cells reflects results observed in vitro; both pp78 and gp57Tac are phosphorylated in PBL-T and Jurkat, but not in U937. These data demonstrate that the IL 2 receptor complex contains an IL 2 responsive protein kinase activity and may signal the cell through a phosphorylation event. PMID- 3040858 TI - Regulation of HLA class II antigen expression: intracellular signaling molecules responsible for the regulation by IFN-gamma and cross-linking of Fc receptors in HL-60 cells. AB - The cross-linking of Fc receptors (FcR) on HL-60 cells inhibited the ability of recombinant IFN-gamma to induce HLA class II antigens. This appeared to be correlated with intracellular mRNA level. HL-60 lacked detectable HLA class II mRNA. IFN-gamma led to appearance of these transcripts, which were canceled by the cross-linking of FcR. Therefore, experiments were designed to investigate the intracellular signaling molecules regulating the appearance of HLA class II molecules or transcripts. The expression of HLA class II antigen induced by IFN gamma was blocked by a calmodulin antagonist, W-7, but not by a protein kinase C (PKC) inhibitor, H-7. Furthermore, a direct activator of PKC, phorbol myristate acetate, was not able to induce the HLA class II antigen expression. These results suggest that IFN-gamma induces HLA class II antigens on HL-60 cells via a calcium-calmodulin pathway and not via a PKC pathway. Calmodulin is activated by a transient rise in the cytosolic free calcium. In fact, the measurement of calcium influx into HL-60 cells showed that a remarkable and time-dependent calcium accumulation was caused by IFN-gamma, and that depletion of Ca2+ from culture medium resulted in failure of IFN-gamma to induce class II antigen expression. Furthermore, calcium ionophore, A23187, by itself induced HLA class II antigen expression. These results suggest that IFN-gamma stimulates calcium influx and activates the calmodulin branch of the calcium messenger system, resulting in the induction of class II antigen expression on HL-60 cells. On the other hand, cross-linking of FcR elicited the accumulation of intracellular cAMP, which appeared to suppress the IFN-gamma-induced calcium influx, resulting in annulling HLA class II antigen-inducing activity of IFN-gamma. These intracellular events of HL-60 regulate the expression of HLA class II transcripts and molecules. PMID- 3040859 TI - Polypeptide fragments of horse cytochrome c activate a small subset of secondary B lymphocytes primed against the native protein. AB - Horse cytochrome c (cyt c) and two large, overlapping cyanogen bromide-cleaved fragments (1-80 and 66-104), together encompassing the entire length of the polypeptide chain, were examined for their abilities to stimulate into antibody production individual secondary B lymphocytes primed against the intact protein. T cell help was provided against the carrier protein, hemocyanin, to which cyt c and its peptides were conjugated by using glutaraldehyde. All the B cells activated by both of the fragments elicited antibodies that reacted with intact cyt c in enzyme-linked immunosorbent assay, whereas only a fraction of the antibodies elicited by the intact protein reacted with the peptides. However, in general, antibodies reactive with the polypeptide fragments, whether elicited by the intact protein or by the fragments, could not be effectively inhibited from binding plate-bound cyt c in enzyme-linked immunosorbent assay in the presence of soluble native cyt c. This indicates that these antibodies are specific for denatured forms of cyt c that apparently arise during the chemical coupling of cyt c to carrier molecules for immunization and/or during emulsification of the immunogen in adjuvant. Whereas, at most, 5% of the secondary B cells specific for native cyt c could be activated by the 1-80 fragment, even fewer were activated by the 66-104 fragment. Therefore, it is unlikely that smaller peptides which fail to assume native conformation would be effective. Antibodies elicited in vivo in a primary response to the 1-80 fragment also failed to bind native cyt c. These results suggest that linear peptides intended to mimic epitopes on globular proteins, and which have not been engineered to adopt native conformation, will not be very effective either as primary or as secondary vaccines for B cell activation. PMID- 3040860 TI - Trace levels of bacterial lipopolysaccharide prevent interferon-gamma or tumor necrosis factor-alpha from enhancing mouse peritoneal macrophage respiratory burst capacity. AB - Preexposure of resident mouse peritoneal macrophages for 1 hr to traces of bacterial lipopolysaccharide (LPS) (less than or equal to 1 ng/ml) rendered the cells refractory to activation by recombinant interferon-gamma (rIFN gamma) or recombinant tumor necrosis factor-alpha (rTNF alpha), as evaluated by release of H2O2 upon stimulation with phorbol myristate acetate. Inhibition persisted for at least 4 days. Fifty percent inhibition of activation mediated by rIFN gamma followed 1 hr exposure to 10 pg/ml LPS. Fifty percent inhibition of activation mediated by rTNF alpha was achieved with 1 hr exposure to 1 pg/ml LPS. Such low levels LPS exposures (concentration X time) are far below those reported for many other actions of LPS on host cells. Inhibition was partially prevented by the cyclooxygenase inhibitors indomethacin, ibuprofen, and acetylsalicylic acid. Exogenous prostaglandins PGE1 and PGE2, and the 3',5'-cyclic adenosine monophosphate analog dibutyryl cyclic adenosine monophosphate (cAMP), mimicked the inhibitory effect of LPS in a dose-dependent manner, consistent with the hypothesis that formation of endogenous cyclooxygenase products in response to LPS may elevate intracellular cAMP and that the latter may mediate the observed inhibition. In addition, neutralizing antibody against IFN alpha and IFN beta selectively prevented LPS inhibition of activation mediated by rIFN gamma, but not by rTNF alpha. This suggests that IFN alpha and/or IFN beta induced by LPS also contributed to inhibition of activation by rIFN gamma. Thus, release of LPS may afford microorganisms a means by which to interfere with immunologically mediated enhancement of the respiratory burst-dependent antimicrobial capacity of macrophages. PMID- 3040861 TI - Correlation between the lipopolysaccharide response of mice and the capacity of mouse peritoneal cells to transfer an antiviral state. Role of endogenous interferon. AB - Freshly harvested mouse peritoneal cells, from normal lipopolysaccharide (LPS) responsive (Lpsn) mice, were capable of transferring an antiviral state (to vesicular stomatitis virus) to "in vitro aged" mouse macrophages permissive for viral replication. The transfer of the antiviral state was completely abrogated by addition of antibody to interferon (IFN)-alpha/beta in the co-culture medium. In contrast, even large numbers of donor peritoneal cells from LPS-hyporesponsive (Lpsd) C3H/HeJ and C57BL/10ScCR mice did not transfer an antiviral state to target cells. Although peritoneal macrophages from Lpsd mice did not transfer an antiviral state to target cells, they were nevertheless found to be in an antiviral state when first placed in culture. Injection of mice with antibody to mouse IFN-alpha/beta rendered peritoneal macrophages from both Lpsn and Lpsd mice permissive for vesicular stomatitis virus. The decay of this initial antiviral state in peritoneal macrophages during in vitro culture was far more rapid for Lpsd mice than for normal mice. Addition of antibody to mouse IFN-alpha/beta markedly enhanced the in vitro decay of the antiviral state of peritoneal macrophages. Treatment of total peritoneal cells from Lpsn mice with LPS resulted in IFN production, whereas IFN was not detected in the cellfree medium of LPS treated peritoneal cells from Lpsd C3H/HeJ and C57BL/10ScCR mice. Genetic studies with F1 hybrids between Lpsn and Lpsd mice and with Lpsn and Lpsd recombinant inbred strains revealed a striking correlation between the capacity of peritoneal cells to transfer an antiviral state and their capacity to produce IFN after stimulation with LPS, suggesting that closely linked, if not identical, genes are in some way involved in the transfer of antiviral state as well as in the LPS response by peritoneal cells of normal mice. PMID- 3040862 TI - In vitro human antibody production to the Haemophilus influenzae type b capsular polysaccharide. AB - In vitro production of human antibody to the Haemophilus influenzae type b capsular polysaccharide (PRP) and to tetanus toxoid (TT) and diphtheria toxoid was measured in culture supernatants of peripheral blood mononuclear cells and by enumeration of antibody secreting cells (AbSC) in an enzyme-linked immunosorbent plaquing assay. Normal adult peripheral blood mononuclear cells stimulated with Epstein-Barr virus secreted anti-PRP antibody with a frequency of 1/552 to 1/1190 relative to total Ig secreting cells; the frequency of AbSC to tetanus toxoid (TT) was 7.5 times higher (p less than 0.05). These frequencies did not change significantly after in vivo immunization, although the isotype distribution shifted toward increased IgG for TT and increased IgG and IgA for PRP. At 8 days postimmunization, spontaneous AbSC to PRP and TT were detected; frequencies for total anti-TT AbSC again being higher than anti-PRP, but there were significantly more IgA plaques among anti-PRP AbSC. Spontaneous AbSC were suppressed in culture by pokeweed mitogen and enhanced by cyclosporine. Three wk after in vivo immunization with PRP and TT, in vitro stimulation with pokeweed mitogen, Staphylococcus aureus Cowan 1 bacteria, or antigen induced anti-TT but not anti PRP in vitro antibody secretion, although Epstein-Barr virus induced both. These data suggest that PRP, a polysaccharide, and TT, a protein, differ in their requirements for in vitro activation with antigen and mitogens. PMID- 3040863 TI - Effects of anti-proliferative cyclic AMP on interleukin 2-stimulated gene expression. AB - Elevation of intracellular cyclic adenosine 3':5' monophosphate (cAMP) inhibits interleukin 2 (IL-2)-stimulated proliferation of a murine cytotoxic cell clone, CT6. The effects of antiproliferative dosages of stable cAMP-derivative, 8 bromoadenosine 3':5'-monophosphate (8-Br-cAMP), on steady state mRNA expression stimulated by IL-2 was examined. IL-2 stimulated mRNA accumulation of three nuclear proto-oncogenes c-fos, c-myc, and c-myb. 8-Br-cAMP alone stimulated c fos, c-myb, and IL-2 receptor mRNA accumulation as determined by Northern blot analysis. 8-Br-cAMP, however, markedly inhibited c-myc expression stimulated by IL-2. Furthermore, although c-fos and IL-2 receptor mRNA expression was potentiated by 8-Br-cAMP, suppression of protein synthesis was seen. We show that antiproliferative cAMP stimulates similar mRNA expression as does IL-2, with the exception of c-myc. Although a comparative stimulation of steady state mRNA accumulation of some genes occurs, cAMP may profoundly effect protein synthesis. cAMP, therefore, acts on multiple targets involved in the macromolecular events stimulated by IL-2. PMID- 3040864 TI - Biochemical characterization and biosynthesis of the Ki-1 antigen in Hodgkin derived and virus-transformed human B and T lymphoid cell lines. AB - The Hodgkin-associated Ki-1 antigen was analyzed in different cell lines. In Hodgkin analogous L428 cells, biosynthetically labeled with radioactive amino acids, the Ki-1 antibody precipitated three glycoproteins with 90, 105, and 120 kDa, respectively. Surface-labeling revealed that the two larger components were membrane-associated forms of the Ki-1 antigen, although the 90-kDa molecule was shown in pulse-chase experiments to be the precursor of the 105- and 120-kDa forms. All three forms of the Ki-1 antigen possess a tunicamycin-sensitive 6-kDa N-linked carbohydrate moiety. O-Linked oligosaccharides could not be detected. Thus, the differences in m.w. are probably not due to glycosylation. The ionophore monensin prevented the appearance of the membrane-associated molecules, which demonstrated that they are assembled between the transcompartment of the Golgi complex and their insertion into the cell membrane. The 90-kDa precursor molecule cannot be generated by disulfide reduction from the two larger forms. After internal labeling with P-32, only the 105- and 120-kDa bands became visible, indicating that the Ki-1 molecule is phosphorylated after its processing into the two larger membrane-associated forms. Analysis of the Ki-1 antigens from other cell lines demonstrated that after external labeling of two other Hodgkin derived cell lines, six Epstein-Barr virus lymphoblastoid cell lines and one human T leukemia virus I-positive T cell line, both the 105- and the 120-kDa membrane molecules could be detected, regardless of the presence or type of virus integrated. PMID- 3040865 TI - Detection of ecto-5'-nucleotidase on rat B and T lymphocyte subpopulations using a monoclonal antibody in rosetting reactions. AB - A mouse monoclonal antibody to rat 5'-nucleotidase (5N 4-2 McAb) was used in the direct anti-determinant rosetting reaction (DARR) to demonstrate the ecto-5' nucleotidase molecule in preparations of rat lymphocytes. Results indicated that 35.5 +/- 7.5% of peripheral blood lymphocytes (PBL), 37.3 +/- 4.8% of lymph node cells (LN) and 37.0 +/- 8.5% of spleen lymphocytes expressed the 5N 4-2 antigen. Depletion studies and mixed rosetting reactions (MRR) showed that the 5N 4-2 antigen was mainly expressed on rat T lymphocytes rather than on B lymphocytes: In fact 59.6 +/- 3.2% (in PBL), 76.5 +/- 0.6% (in LN) and 67.1 +/- 1.3% (in spleen) of T lymphocytes exhibited the 5N 4-2 antigen compared to only 26.5 +/- 2.6% (in PBL), 34.0 +/- 2.1% (in LN) and 46.1 +/- 12.0% (in spleen) of B lymphocytes. As expected a strong association was found between the expression of 5N 4-2 antigen and 5'-nucleotidase enzyme activity on lymphocytes. Both 5N 4-2 positive cells and enzyme activity were preferentially exhibited in the T lymphocyte subpopulation, and 92% of the enzyme activity was observed in a 5N 4-2 antigen positive subpopulation. PMID- 3040866 TI - The isolation and characterization of enriched microvascular endothelial cells from mouse adipose tissue. The induction of class II molecules of the major histocompatibility complex (MHC) by interferon-gamma (IFN-gamma). AB - Collagenase digestion and selective filtration have been used to establish primary cultures from mouse omentum of cells enriched for microvascular endothelium. In culture these cells exhibit a cobblestone morphology characteristic of endothelial cells, metabolize an acetylated low-density lipoprotein, and exhibit trace levels of angiotensin-converting enzyme activity. In primary cultures, the cells are negative for class II molecules (Ia) of the major histocompatibility complex (MHC) but can be induced to express these molecules by exposure to a supernatant from concanavalin A (ConA)-treated rat spleen cells or by recombinant interferon-gamma (rIFN-gamma). This procedure can provide a readily available source of enriched endothelial cells which can be used to understand the interactions between these cells and cells of the immune system. PMID- 3040867 TI - Two phenotypically distinct T cells are involved in ultraviolet-irradiated urocanic acid-induced suppression of the efferent delayed-type hypersensitivity response to herpes simplex virus, type 1 in vivo. AB - When UVB-irradiated urocanic acid, the putative photoreceptor/mediator for UVB suppression, is administered to mice it induces a dose-dependent suppression of the delayed-type hypersensitivity response to herpes simplex virus, type 1 (HSV 1), of similar magnitude to that induced by UV irradiation of mice. In this study, the efferent suppression of delayed-type hypersensitivity by UV-irradiated urocanic acid is demonstrated to be due to 2 phenotypically distinct T cells, (Thy1+, L3T4-, Ly2+) and (Thy1+, L3T4+, Ly2-). The suppression is specific for HSV-1. This situation parallels the generation of 2 distinct T-suppressor cells for HSV-1 by UV irradiation of mice and provides further evidence for the involvement of urocanic acid in the generation of UVB suppression. PMID- 3040868 TI - Effect of 9-(1,3-dihydroxy-2-propoxymethyl)guanine and recombinant human beta interferon alone and in combination on simian varicella virus infection in monkeys. AB - Treatment of viral infections with combinations of antiviral agents may permit administration of reduced doses of either or both drugs. Lowered doses may reduce associated toxicity. Intravenous administration of substantial doses of either human recombinant beta interferon (rHuIFN-beta) or 9-(1,3-dihydroxy-2 propoxymethyl)guanine (DHPG) prevents development of simian varicella virus infection in African green monkeys. Daily doses of 2 X 10(6) U of rHuIFN-beta/kg inhibited clinical disease in monkeys inoculated with simian varicella virus, and doses of DHPG between 20 and 60 mg/kg per day were necessary for similar antiviral effects. Intravenous administration of combinations of rHuIFN-beta and DHPG permitted an approximately 100-fold reduction in the effective dose of rHuIFN-beta and a 10-fold reduction in the effective dose of DHPG. Analysis of data relating to viremia by using the method of the median-effect principle showed the combination of rHuIFN-beta and DHPG was strongly synergistic in treatment of this infection. PMID- 3040870 TI - Efficacy of cytomegalovirus immunoglobulin in marrow transplant recipients with cytomegalovirus pneumonia. PMID- 3040869 TI - Antibody response to virus-encoded proteins after cytomegalovirus mononucleosis. AB - We determined serial IgG antibody responses to cytomegalovirus (CMV)-encoded proteins in sera collected over a one-year interval from 14 subjects with CMV mononucleosis. Antigens from infected human fibroblasts included three components: cytoplasmic, nuclear, and high-speed pellet. Antibody was detected by radioimmunoprecipitation followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Twenty to 21 bands were observed with the cytoplasmic component, whereas 10 and 9, respectively, were seen with the nuclear and high-speed pellet antigens. The most intense reactions occurred with the higher-molecular-mass proteins (50-215 kDa) by using the cytoplasmic and high-speed antigens and with the more rapidly migrating proteins (less than 50 kDa) by using the nuclear antigen. The precipitin responses increased for three months or more after onset of symptoms with the nuclear and high-speed pellet antigens but peaked within one to two months with the cytoplasmic antigen. PMID- 3040871 TI - Infection of vervet monkey bone marrow stromal cells with simian cytomegalovirus. PMID- 3040872 TI - [A case report of primary malignant fibrous histiocytoma of the lung with metastasis to the stomach]. PMID- 3040874 TI - Intraovarian markers of follicular and oocyte maturation. AB - The use of ovulation induction for multiple follicular growth in in vitro fertilization (IVF) has introduced the problem of follicular asynchrony. As a consequence of the asynchrony, the parameters most commonly used by IVF groups to assess follicular and oocyte quality within those follicles are not sufficiently sensitive or specific. Thus, each follicle must be considered separately, and specific markers of follicular and/or oocyte maturation must be sought from within the follicle. In this review we analyze previous reports of potential markers of follicular and oocyte maturation. In regards to the follicular fluid constituents, the level of estradiol in follicular fluid correlates with fertilization and pregnancy in stimulated cycles. Other steroids are only helpful when specific stimulation protocols are used. The level of some follicular proteins such as alpha-1-antitrypsin and fibrinogen also correlates with fertilization and pregnancy outcome. Cyclic AMP levels in follicular fluid are significantly reduced in follicles leading to conception. Regulators of oocyte maturation, such as the Oocyte Maturation Inhibitor (OMI) or the Meiosis Inducing Substance (MIS) have also been correlated with IVF outcome, but their exact structure remains still unknown. In addition, other sophisticated parameters, such as chemotactic activity of human leukocytes, or simple methods, such as the presence of intrafollicular echoes, have also been used as successful markers in predicting IVF outcome. PMID- 3040873 TI - Organization of the human class I major histocompatibility complex genes. PMID- 3040875 TI - Mixed familial polyposis syndromes. AB - Three cases of polyposis consisting of adenomatous and hamartomatous elements are described. The literature on mixed polyposis syndromes is reviewed. PMID- 3040876 TI - Chemotactic factor binding and functional capacity: a comparison between human granulocytes and differentiated HL-60 cells. AB - In the presence of dimethyl sulfoxide (DMSO), the leukemic promyelocytic cell line HL-60 will differentiate into mature polymorphonuclear granulocytes. In the present report, we compare chemotactic factor binding and function in HL-60 cells with that of normal human granulocytes using the chemotactic peptide N formylmethionyl-leucyl-phenylalanine (FMLP) as ligand. The cellular response measured as CL was changed as a result of storage or conditioning of normal peripheral blood cells. With these cells, a conditioning procedure at room temperature resulted in a pronounced increase in the CL response. The increase of the CL response was probably a result of increased expression of cryptic receptors, since the changes of the oxidative response to FMLP was accompanied by increased binding of the peptide to the cell surface. Scatchard analysis revealed that the increased binding was due to an increased number of receptors. In differentiated HL-60 cells, conditioning neither led to increased production of oxidative metabolites, nor to any increased binding of the peptide. The data thus indicate that many FMLP receptors could reside in a cryptic site that is not accessible to extracellular ligands, and that conditioning results in an increased exposure of these receptors, followed by an increased oxidative response to the ligand in normal cells but not in mature HL-60 cells. PMID- 3040877 TI - Arachidonic and eicosapentaenoic acid metabolism in bovine neutrophils and platelets: effect of calcium ionophore. AB - Substitution of dietary fatty acids has potential for altering the inflammatory response. The purpose of the present study was to define the metabolites of arachidonic acid (AA) and eicosapentaenoic acid (EPA) secreted by bovine peripheral blood neutrophils and platelets. High performance liquid chromatography was used to characterize cyclooxygenase and lipoxygenase metabolites secreted in response to the calcium ionophore A23187. Cells were prelabelled with 3H-AA or 3H-EPA prior to challenge with the calcium ionophore. Bovine neutrophils secreted leukotriene B4 (LTB4) and 5-hydroxyeicosatetraenoic acid (5-HETE) as the major metabolites of AA, as well as the corresponding leukotriene B5 (LTB5) and 5-hydroxyeicosapentaenoic acid (5-HEPE) metabolites of EPA. Peptidoleukotrienes derived from 3H-AA or 3H-EPA were not detected under these conditions. The major tritiated metabolites secreted from bovine platelets were: thromboxane A2, measured as the stable metabolite thromboxane B2 (TXB2); hydroxyheptadecatrienoic acid (HHT) and 12-HETE derived from 3H-AA; and the omega 3 analogs TXB3 and 12-HEPE, derived from 3H-EPA. Preferred substrate specificities existed amongst the AA- and EPA-derived metabolites for the intermediary enzymes involved in the arachidonic acid cascade. These findings support the hypothesis that substitution of membrane-bound AA by EPA has potential for modulation of the host inflammatory response following cellular phospholipid mobilization. PMID- 3040878 TI - Platelet-activating factor stimulates metabolism of phosphoinositides via phospholipase A2 in primary cultured rat hepatocytes. AB - Addition of platelet-activating factor (PAF) to cells doubly labeled with [14C]glycerol plus [3H]arachidonic acid resulted in a transient decrease of [14C]glycerol-labeled phosphatidylinositol (PI) and a transient increase of [14C]glycerol-labeled lysophosphatidylinositol (LPI). [3H]Arachidonate-labeled PI, on the other hand, decreased in a time-dependent manner. The radioactivity in phosphatidylethanolamine, phosphatidylcholine, sphingomyelin, and phosphatidylserine did not change significantly. The 3H/14C ratio decreased in PI in a time-dependent manner, suggesting the involvement of a phospholipase A2 activity. Although PAF also induced a gradual increase of diacylglycerol (DG), the increase of [14C]glycerol-labeled DG paralleled the loss of triacyl [14C]glycerol and the 3H/14C ratio of DG was 16 times smaller than that of PI. Thus, DG seemed not to be derived from PI. In myo- [3H]inositol-prelabeled cells, PAF induced a transient decrease of [3H]phosphatidylinositol-4,5-bis-phosphate (TPI) and [3H]phosphatidylinositol-4-phosphate (DPI) at 1 min. PAF stimulation of cultured hepatocytes prelabeled with 32Pi induced a transient decrease of [32P]polyphosphoinositides at 20 sec to 1 min. [32P]LPI appeared within 10 sec after stimulation and paralleled the loss of [32P]PI. [3H]Inositol triphosphate, [3H]inositol diphosphate, and [3H]inositol phosphate, which increased in a time dependent manner upon stimulation with adrenaline, did not accumulate with the stimulation due to PAF. These observations indicate that PAF causes degradation of inositol phospholipids via phospholipase A2 and induces a subsequent resynthesis of these phospholipids. PMID- 3040879 TI - Transmission of AIDS by insects? PMID- 3040880 TI - Simultaneous bilateral Wilms' tumors. A long-term survivor with histologic maturation. PMID- 3040881 TI - Vasopressin receptors influencing the secretion of ACTH by the rat adenohypophysis. AB - The effects of selective agonists and antagonists of type 1 (V1) and type 2 (V2) vasopressin receptors on the secretion of ACTH in vitro by segments of adenohypophysial tissue and in vivo in rats pretreated with pentobarbitone and chlorpromazine were studied in the presence and absence of the 41 amino acid containing peptide, corticotrophin-releasing factor-41 (CRF-41). The non selective vasopressin receptor agonist, arginine vasopressin (AVP) and the V1 receptor agonist, felypressin caused dose-related increases in ACTH release in vivo and in vitro but the V2-receptor agonist, desmopressin was only weakly active in this respect. Their actions in vitro were antagonized competitively by the V1-receptor antagonist, d(C2H5)2-AVP, but were unaffected by the V2-receptor antagonist, d(CH2)5-D-Iso2-Thr4-AVP. Arginine vasopressin, felypressin and desmopressins in concentrations considerably lower than those necessary to elicit directly the release of ACTH, potentiated, in a dose-related manner, the activity of CRF-41 in vitro. The potentiating effects were not antagonized by the V2 receptor antagonist or by low concentrations of the V1-receptor antagonist. At a higher concentration, the V1-receptor antagonist reduced, but did not abolish, the potentiating effects of AVP and its analogues. However, at this concentration, it also exhibited weak intrinsic activity and, like the agonists, potentiated the response to CRF-41. The results suggest that the direct effect of AVP on ACTH release is mediated by V1-like receptors. The vasopressin receptors involved in the potentiation of CRF-41 activity appear to be different. PMID- 3040882 TI - Injection of antibodies into the nucleus of amphibian oocytes: an experimental means of interfering with gene expression in the living cell. PMID- 3040883 TI - Amino acids at the site of V kappa-J kappa recombination not encoded by germline sequences. AB - Murine V kappa-J kappa recombination is characterized by a maintenance of size at the site of recombination and the use of nucleic acids found only in germline sequences. This is in contrast to heavy chain VH-D-JH assembly where random nucleotides are added at the recombination sites to produce considerable size variation, even though the heptamer/nonomer recombination sequences are identical in both kappa and heavy chain genes. We have examined the origin of an unusual amino acid, Ile, found at the site of V kappa-J kappa recombination in antigalactan antibodies, by sequence analysis of the corresponding rearranged and germline genes. Results indicate that the Ile codon can be generated by use of a single nucleotide 3' of the V kappa segment in combination with the second and third nucleotides of the first codon of J kappa 5 or J kappa 4. However, several antigalactan antibodies express Ile in combination with J kappa 2. An Ile codon cannot be generated by recombination in any reading frame between germline V kappa and J kappa 2 segments. These results suggest that the origin of the Ile codon in lines using J kappa 2 may represent a novel even in murine light chain assembly, possibly similar to the de novo addition of nucleotides observed in heavy chain gene recombination. PMID- 3040884 TI - Cytolytic T lymphocyte recognition of the murine cytomegalovirus nonstructural immediate-early protein pp89 expressed by recombinant vaccinia virus. AB - The murine immediate-early (IE) protein pp89 is a nonstructural virus-encoded phosphoprotein residing in the nucleus of infected cells, where it acts as transcriptional activator. Frequency analysis has shown that in BALB/c mice the majority of virus-specific CTL recognize IE antigens. The present study was performed to assess whether pp89 causes membrane antigen expression detected by IE-specific CTL. Site-directed mutagenesis has been used to delete the introns from gene ieI, encoding pp89, for subsequent integration of the continuous coding sequence into the vaccinia virus genome. After infection with the vaccinia recombinant, the authentic pp89 was expressed in cells that became susceptible to lysis by an IE-specific CTL clone. Priming of mice with the vaccinia recombinant sensitized polyclonal CTL that recognized MCMV-infected cells and transfected cells expressing pp89. Thus, a herpesviral IE polypeptide with essential function in viral transcriptional regulation can also serve as a dominant antigen for the specific CTL response of the host. PMID- 3040885 TI - Transient rearrangements of the T cell antigen receptor alpha locus in early thymocytes. AB - The dull Ly-1 double-negative (Ly-1dull, Lyt-2-, L3T4-) subpopulation appears to be the major precursor group of T lymphocytes in the thymus. In examining the status of the alpha, beta, and gamma chain genes for T cell receptors (TCR) in this population of cells and hybridomas made from them, we find that all of these loci appear to begin DNA rearrangements in a nearly simultaneous fashion. In the case of the gamma genes, these involve V gamma----J gamma C gamma gene rearrangements; with the beta chain genes, both D beta----J beta C beta rearrangement and V beta----D beta J beta C beta rearrangements are evident; and in the case of the alpha locus, assayed in part by pulsed-field gel electrophoresis, they take the form of a novel series of rearrangements occurring 80 kb or more 5' to the C alpha gene. These alpha locus rearrangements are well away from any of the J alpha gene segments found in cDNA clones to date and are deleted in most mature thymocytes and functional T cell lines. Therefore they appear to represent a distinct class of rearrangement that occurs before V alpha- --J alpha joining. These distinctions between the character of the TCR gene rearrangements in these cells represent useful markers in further distinguishing different stages of T cell differentiation within this compartment of early T cells. In addition, the unexpected discovery of clonal rearrangements so far away from any of the expressed J alpha gene segments, and at a stage where there is little or no stable C alpha RNA present, has interesting implications for the hierarchy of TCR gene expression. PMID- 3040887 TI - [The angiographic appearance of hepatocellular carcinoma: differences between small and large tumors]. PMID- 3040888 TI - Bleeding jejunal varices in a cirrhotic patient with hepatocellular carcinoma. PMID- 3040886 TI - Effect of tumor necrosis factor alpha on mitogen-activated human B cells. AB - In this study we demonstrate that the monocyte/macrophage product, tumor necrosis factor alpha (TNF-alpha), has significant in vitro effects of B cell function. It costimulated with anti-mu in the induction of B cell DNA synthesis, and it prolonged the DNA synthesis initiated in B cell cultures stimulated with the human B cell mitogen, Staphylococcus aureus Cowan strain I (SAC). The addition of either IL-1 or IFN-gamma to TNF-alpha resulted in a substantial further increase in DNA synthesis. The addition of TNF-alpha to IL-2, a known inducer of SAC activated B cell Ig secretion, resulted in a twofold enhancement in the amount of IL-2 stimulated B cell Ig secretion. Receptor binding studies with 125I-TNF-alpha demonstrate a marked increase in TNF-alpha binding sites after B cell activation (approximately 6,000 sites per cell, with an apparent Kd of 2.0 X 10(-10) M). Thus, TNF-alpha may be an important factor in human B cell function and is likely to interact with other T cell and monocyte derived cytokines in the regulation of human B cell proliferation and Ig production. PMID- 3040889 TI - The arcABC operon required for fermentative growth of Pseudomonas aeruginosa on arginine: Tn5-751-assisted cloning and localization of structural genes. AB - Pseudomonas aeruginosa is able to utilize L-arginine as the energy source for growth under anaerobic, nitrate-free conditions. Mutations in the chromosomal arcABC gene cluster specifying the inducible arginine deiminase pathway enzymes abolish fermentative growth on arginine. From two different arc::Tn5-751 insertion mutants of P. aeruginosa recombinant plasmids have been derived which carry a resistance marker of transposon Tn5-751 plus flanking parts of the arc region. These recombinant plasmids served to reconstruct in vitro the functional arcABC cluster on a 5.6 kb fragment, which was inserted into the broad-host-range vector pKT240. In P. aeruginosa this 5.6 kb segment complemented arcABC mutations in trans and contained the control region necessary in cis for arc enzyme induction by oxygen limitation and arginine. The results of subcloning experiments and transcriptional lacZ fusions, the polarity of transposon insertions and the effect of external promoters led to the conclusion that the structural genes arcA (for arginine deiminase), arcB (for catabolic ornithine carbamoyltransferase) and arcC (for carbamate kinase) are contiguous and transcribed in the same direction. Thus, the arcABC cluster appears to have the characteristics of an operon. In Escherichia coli the cloned arcABC genes were expressed at low, non-inducible levels; strong vector promoters enhanced arc expression up to 100-fold. This indicates that transcriptional initiation at the arc promoter(s) is poor in E. coli. PMID- 3040890 TI - Methylation of spore DNA in Bacillus coagulans strain 26. AB - The modification status of DNA throughout the life cycle of Bacillus coagulans strain 26 was analysed by restriction analysis with methylation-sensitive enzymes. A significant fraction of the GATC sequences (dam target) in spore DNA contain N6-methyladenine, a modification that is lacking during the vegetative phase. From the modulation of the modification pattern of GATC sites, the existence of a de novo methylase may be inferred. Spore DNA was more sensitive than vegetative cell DNA to BamHI, HpaI, SalI and XhoI, indicating that the sites for these enzymes are modified during the vegetative growth phase. PMID- 3040891 TI - Restriction mapping and close relationship of the DNA of Streptomyces erythraeus phages 121 and SE-5. AB - The biological properties and genome structure of two actinophages, 121 and SE-5, infecting Streptomyces erythraeus were characterized. They had the same host range (limited to S. erythraeus) and similar DNA G + C contents (around 60 mol %). Restriction maps of their genomes also showed many similarities. The close relationship between the two phages was confirmed by DNA hybridization experiments: large parts of their genomes were homologous, except for a segment in the middle of the map, where no hybridization was detected. PMID- 3040892 TI - Inhibition of restriction in Streptomyces clavuligerus by heat treatment. AB - Inefficient transformation of Streptomyces clavuligerus protoplasts by DNA from the plasmid pIJ702, isolated from S. lividans, was attributed to restriction in view of the observation that efficient transformation was observed using modified pIJ702 (isolated from S. clavuligerus). The restriction system could be partially inhibited by treating protoplasts at 45 degrees C prior to transformation. This treatment increased the transformation frequencies of pIJ702 DNA by 100-fold and was used to introduce other plasmids into S. clavuligerus. PMID- 3040893 TI - Factors enhancing genetic transformation of intact yeast cells modify cell wall porosity. AB - Genetic transformation of intact cells of Saccharomyces cerevisiae, achieved by incubating the cells with plasmid DNA in the presence of PEG, could be enhanced, not only by pretreatment of the cells with Li+ and 2-mercaptoethanol, as has been reported previously, but also by pretreatment with proteolytic enzymes. The efficiency of transformation with 2-mercaptoethanol rose dramatically when the pretreated cells had been handled in osmotically stabilized media. Following all the pretreatments the cells became leaky for nucleic acids as detected by the presence of endogenous RNAs in the medium. The pretreatments evidently facilitated the passage of transforming DNA across the cell wall. PMID- 3040894 TI - Mutations affecting gluconate catabolism in Escherichia coli. Genetic mapping of the locus for the thermosensitive gluconokinase. AB - An Escherichia coli strain unable to use gluconate was isolated by spontaneous curing of lambda cI857 s7 xis6 b515 b519, lambda cI857 s7 delta(A-att) dargI valS lysogens. Two lesions, linked to asd and pyrB markers, respectively, were necessary to produce this phenotype. The asd-linked mutation gnt-17, of regulatory type, seems to affect the expression of the major system of gluconate utilization (min 75) as well as that of 6-phosphogluconate dehydratase (gene edd, min 41), the first enzyme of the Entner-Doudoroff pathway. A closely linked suppressor of gnt-17 causes constitutivity of these activities; this suppressor resembles gntR, which is also in the asd region. Hence, it is possible that gnt 17 is a super-repressing allele of gntR, rather than a positive controlling element. Lesion gnt-17 alone does not prevent the utilization of gluconate; for this, the mutation gnt-18 at 96.9 min is also necessary. This mutation abolishes the thermosensitive gluconokinase activity and thus eliminates the subsidiary ability to catabolize gluconate. Accordingly, gnt-18 seems to be allelic with gntV, the locus postulated as being in the pyrB region specifying the thermosensitive gluconokinase. PMID- 3040895 TI - Virus-specific serum IgG, IgM, and IgA antibodies in cytomegalovirus mononucleosis patients as determined by immunoblotting technique. AB - The immune response to individual human cytomegalovirus (CMV) structural polypeptides was studied in paired sera from 15 adult CMV mononucleosis (CMV-MN) patients and healthy controls by immunoblotting technique (IB). IgM and IgG antibodies to at least 11 structural polypeptides with molecular weights of 28K, 49K, 55K, 57K, 66-70K, 82K, 87K, 110K, 150K, 205K, and 235K were detected in the patients' sera in the serum sample obtained in the acute phase of the disease. IgA antibodies to polypeptides with molecular weights of 66-70K, 82K, 110K, and 150K were also detected in these sera. In healthy seropositive adults, IgG antibodies with the same molecular weight polypeptides, excluding the 205K and 235K polypeptides, were detected as in convalescent CMV-MN patients. A prominent reactivity of IgM and IgA antibodies to the 66-70K and 150K polypeptides was noted in the acute sera from all the CMV-MN patients examined, but not in a number of late convalescent sera. The potential implications of these findings in the development of specific serological tests are discussed. PMID- 3040896 TI - Production and characterisation of a human monoclonal antibody to cytomegalovirus and its use in an early nuclear fluorescence assay. AB - A human monoclonal antibody to cytomegalovirus (CMV) was produced by transforming peripheral blood mononuclear cells of a patient with recent CMV infection. It is directed against a late antigen located in the nucleus of CMV infected fibroblasts at 24-72 hours postinfection and immuneprecipitates 65K and 48K proteins from 35S-labelled CMV infected cells. Results of its use in an early nuclear fluorescence assay for rapid diagnosis are presented. PMID- 3040897 TI - Effect of polymorphonuclear depletion on experimental Argentine hemorrhagic fever in guinea pigs. AB - The role that polymorphonuclear leukocytes (PMN) may play in Argentine hemorrhagic fever (AHF), an endemo-epidemic disease caused by Junin virus (JV), was investigated in experimentally infected guinea pigs depleted of PMN by means of specific antiserum. In leucopenic animals the evolution of the infection with a highly pathogenic strain of JV was more severe, with earlier mortality and higher virus yields in blood and viscera. The pathological study showed similar lesions in both the control and PMN-depleted animals with the exception of the lung, which showed the pathological picture of the human "pulmonary distress syndrome of the adult" in nontreated guinea pigs and appeared histologically unaltered in the PMN-depleted animals. On the basis of these results it is suggested that in AHF, PMN play a dual role. In the first stage of infection they display a defensive antiviral action, but later on they participate in the pathogenesis of tissue damage. PMID- 3040898 TI - Immunization of monkeys with varicella-zoster virus glycoprotein antigens and their response to challenge with simian varicella virus. AB - African green monkeys (Cercopithecus aethiops) were immunized with three intramuscular injections of gpI, gpII, or gpIII glycoprotein antigens of varicella-zoster virus (VZV). Antibody responses to VZV were determined by enzyme linked immunosorbent assay (ELISA) and to simian varicella virus (SVV) by immunofluorescence and by serum neutralization assays. Two weeks following the third immunization with VZV glycoproteins, the monkeys were challenged by inoculation of SVV. Antibodies to gpII or gpIII partially prevented infection by SVV, while the presence of antibodies to gpI was ineffective in preventing disease induced by SVV challenge. Factors affecting the immunogenicity of these antigens in this model are discussed. PMID- 3040900 TI - Absence of HTLV I from New Zealand. AB - Sera from 1,943 individuals from Auckland, New Zealand, were tested for the presence of serum antibodies to human T cell lymphotropic virus I (HTLV I), mainly with an enzyme-linked immunosorbent assay (ELISA) with cell extracts as target antigen. The individuals tested were blood donors and mostly Caucasian, but included indigenous Maoris and representatives of several groups of Pacific islanders now resident in New Zealand. Also included were 37 patients with various hematological malignancies, including seven with T cell leukemias. Although 1% of samples were positive by ELISA, none of these were confirmed as positives by Western blotting. On the basis of these results we consider that it is unlikely that HTLV I infection occurs in Auckland; however, we cannot exclude the possibility that pockets of virus infection may occur in other parts of New Zealand or the South Pacific. PMID- 3040899 TI - First report of an epidemic of diarrhoea in human neonates involving the new rotavirus and biological characteristics of the epidemic virus strain (KMB/R85). AB - An outbreak of diarrhoea in neonates occurred at the nurseries of the Department of Obstetrics of Zhao Tong Regional Hospital, Yunnan Province, from the middle of August to the end of November, 1985. Fifty-one percent of children were affected 2-8 days after birth. The clinical symptoms were mild; patients mainly had diarrhoea and did not vomit. Rotaviruses were detected in 66.7% by RNA PAGE and in 72.7% by EM. The virus strain designated as KMB/R85 had a typical morphology, which was indistinguishable from that of infantile rotaviruses by EM. The viral RNA genome was composed of 11 segments. The buoyant density in CsCl was 1.377 g/cm3. The KMB/R85 strain possessed a hemagglutinin for rhesus monkey erythrocytes. By ELISA, IEM, and HAI, it was found that KMB/R85 strain did not possess the common group antigen shared by group A rotaviruses and was antigenically similar to the Chinese adult diarrhoea rotavirus (serogroup B). PMID- 3040901 TI - Leucocyte hepatitis B virus DNA in acute and chronic hepatitis B virus infection. AB - In the present study we have investigated 53 patients with a spectrum of acute and chronic hepatitis B virus (HBV) infection for the presence of leucocyte HBV DNA with the aid of molecular techniques. HBV-DNA was detected in peripheral blood mononuclear cells of 31 of 45 (69%) of chronic HBsAg carriers and 2 of 8 (25%) patients with acute hepatitis B. Although HBV-DNA was detected more frequently in leucocytes from those HBsAg carriers seropositive for HBeAg (79%), 50% of those with anti-HBe in serum had leucocytes positive for HBV-DNA independent of the presence of serum HBV-DNA. Examination of various leucocyte subpopulations showed the presence of HBV-DNA in polymorphonuclear leucocytes as well as T- and non-T-enriched mononuclear cell fractions. The HBV-DNA identified was predominantly 3.2-kilobase (kb), while higher molecular weight sequences were rarely detected, and lower molecular weight sequences indicative of active viral replication were not observed. These data indicate that although leucocytes do not actively support viral replication, they frequently harbour 3.2-kb HBV-DNA and may act as a reservoir for infection and, more importantly, since leucocytes contaminate several body secretions, may be involved in virus transmission. PMID- 3040902 TI - Regional distribution of high-affinity gamma-[3H]hydroxybutyrate binding sites as determined by quantitative autoradiography. AB - The distribution of high-affinity binding sites for gamma-[3H]hydroxybutyrate in coronal sections of rat brain was studied by quantitative autoradiographic techniques. Binding sites for this naturally occurring substance, which may possibly have a neurotransmitter role, are concentrated in some restricted areas of the brain, particularly in the limbic system. The hippocampus (especially field CA1 of Ammon's horn, at 292 fmol/mg of tissue), septum (72 fmol/mg of tissue), and cortex (frontal, 113 fmol/mg of tissue; parietal, 103 fmol/mg of tissue; cingulate, 114 fmol/mg of tissue; and entorhinal, 134 fmol/mg of tissue) show pronounced labeling with gamma-[3H]hydroxybutyrate. Binding is much lower in caudatus-putamen (50 fmol/mg of tissue), thalamus, and hypothalamus. Caudal parts of the brain (cerebellum, pons, and medulla) are practically devoid of binding sites. These results strongly support a functional role of endogenous gamma hydroxybutyrate in particularly restricted areas of the rat brain. PMID- 3040903 TI - Isolation and partial characterization of a 56,000-dalton phosphoprotein phosphatase from the blood-brain barrier. AB - A 56,000-dalton protein with inherent phosphoprotein phosphatase activity was isolated from porcine brain capillaries. The enzyme is not activated by divalent metal ions but strongly inhibited by zinc ions. As phosphatase inhibitor 2 readily inhibits the enzymatic activity, the protein can be classified as a type I phosphatase. The protein is stable toward protease treatment. Limited digestion with trypsin does not convert the enzyme into an active form of lower molecular weight. The physical and enzymatical properties of the phosphatase exhibit considerable similarities to those of another 56,000-dalton phosphatase derived from rabbit reticulocytes. PMID- 3040904 TI - A Ca2+-dependent protein kinase activity associated with serotonin binding protein. AB - The endogenous phosphorylation of serotonin binding protein (SBP), a soluble protein found in central and peripheral serotonergic neurons, inhibits the binding of 5-hydroxytryptamine (5-HT, serotonin). A protein kinase activity that copurifies with SBP (SBP-kinase) was partially characterized and compared with calcium/calmodulin-dependent protein kinase II (CAM-PK II). SBP itself is not the enzyme since heating destroyed the protein kinase activity without affecting the capacity of the protein to bind [3H]5-HT. SBP-kinase and CAM-PK II kinase shared the following characteristics: (1) size of the subunits; (2) autophosphorylation in a Ca2+-dependent manner; and (3) affinity for Ca2+. In addition, both forms of protein kinase phosphorylated microtubule-associated proteins well and did not phosphorylate myosin, phosphorylase b, and casein. Phorbol esters or diacylglycerol had no effect on either of the protein kinases. However, substantial differences between SBP-kinase and CAM-PK II were observed: (1) CAM enhanced CAM-PK II activity, but had no effect on SBP-kinase; (2) synapsin I was an excellent substrate for CAM-PK II, but not for SBP-kinase; (3) 5-HT inhibited both the autophosphorylation of SBP-kinase and the phosphorylation of SBP, but had no effect on CAM-PK II. These data indicate that SBP-kinase is different from CAM-PK II. Phosphopeptide maps of SBP and SBP-kinase generated by digestion with S. aureus V8 protease are consistent with the conclusion that these proteins are distinct molecular entities. It is suggested that phosphorylation of SBP may regulate the transport of 5-HT within neurons. PMID- 3040905 TI - Calcium/calmodulin-dependent protein kinase II in squid synaptosomes. AB - The Ca2+/calmodulin (CaM)-dependent protein kinase II system in squid nervous tissue was investigated. The Ca2+/CaM-dependent protein kinase II was found to be very active in the synaptosome preparation from optic lobe, where it was associated with the high-speed particulate fraction. Incubation of the synaptosomal homogenate with calcium, calmodulin, magnesium, and ATP resulted in partial and reversible conversion of the Ca2+/CaM-dependent protein kinase II from its calcium-dependent form to a calcium-independent species. The magnitude of this conversion reaction could be increased by inclusion of the protein phosphatase inhibitor NaF or by substitution of adenosine 5'-O-(3 thiotriphosphate) for ATP. When [gamma-32P]ATP was used, proteins of 54 and 58 kilodaltons (kDa) as well as proteins greater than 100 kDa were rapidly 32P labeled in a calcium-dependent manner. Major 125I-CaM binding proteins in the synaptosome membrane fraction were 38 and 54 kDa. The Ca2+/CaM-dependent protein kinase II was purified from the squid synaptosome and was shown to consist of 54- and 58-60-kDa subunits. The purified kinase, like Ca2+/CaM-dependent protein kinase II from rat brain, catalyzed autophosphorylation associated with formation of the calcium-independent form. These studies, characterizing the Ca2+/CaM dependent protein kinase II in squid neural tissue, are supportive of the putative role of this kinase in regulating calcium-dependent synaptic functions. PMID- 3040906 TI - Subcellular fractionation of the longitudinal smooth muscle/myenteric plexus of dog ileum: dissociation of the distribution of two plasma membrane marker enzymes. AB - The distribution of plasma membrane markers, the sodium pump [evaluated as ouabain-sensitive, potassium-stimulated p-nitrophenyl phosphatase (K+-pNPPase)], [3H]saxitoxin binding, and 5'-AMPase, was studied in the subcellular fractions prepared from the homogenates of the longitudinal smooth muscle/myenteric plexus of dog ileum. The K+-pNPPase activity and [3H]-saxitoxin binding were found to be predominantly associated with the synaptosomal fraction as indicated by the high level of these activities in the crude synaptosomal fraction and by the copurification of K+-pNPPase and [3H]saxitoxin binding, but not 5'-AMPase, with several synaptosomal markers during the fractionation of the crude synaptosomal fraction on density gradients. In contrast to the K+-pNPPase activity and [3H]saxitoxin binding, the 5'-AMPase activity was found to be concentrated in the microsomal pellet. Further fractionation of microsomes on density gradient resulted in copurification of 5'-AMPase but not K+-pNPPase or [3H]saxitoxin binding, with other smooth muscle plasma membrane-bound enzymes, such as high affinity Ca2+-ATPase, Mg2+-ATPase, and Ca2+-ATPase. It was concluded that in the longitudinal smooth muscle/myenteric plexus, the sodium pump activity is present in higher density in the neuronal plasma membranes whereas 5'-AMPase activity is concentrated in the smooth muscle plasma membranes. PMID- 3040907 TI - In vitro protein phosphorylation in head preparations from normal and mutant Drosophila melanogaster. AB - We have characterized protein phosphorylation in vitro in subcellular fractions from Drosophila melanogaster heads. Optimal conditions for the incorporation of 32P into proteins, and its dependence on ATP, divalent cations, and cyclic nucleotides have been determined, as well as the effect of inhibitors of ATPase, protein phosphatase, and protein kinase on protein phosphorylation. Among these inhibitors, Zn2+ was found to affect the incorporation of 32P into specific bands and p-hydroxymercuribenzoate was found to be most suited for freezing the activity of both kinases and phosphatases. Cyclic AMP-dependent protein kinase (cAMP-dPK) activity was present in both supernatant (S2) and particulate (P2) fractions, with the majority (60-85%, depending on the homogenization medium) being associated with S2, as determined by phosphorylation of exogenous synapsin I. cAMP-dPK catalyzed the phosphorylation of at least 18 endogenous polypeptides in S2 and at least 10 endogenous polypeptides in P2. These proteins could be classified on the basis of the extent of stimulation of phosphorylation by cyclic nucleotides, dependence on cyclic nucleotide concentration, and rate of phosphorylation. A phosphoprotein of 51 kilodaltons (pp51) was a major component of the S2 and P2 fractions and displayed properties expected from the regulatory subunit of the cAMP-dPK, R-II. A phosphoprotein doublet of approximately 37 kilodaltons (pp37) was stimulated to the largest extent by cAMP in the P2 and S2 fractions. The phosphorylation of several proteins in both fractions was significantly lowered by the mammalian Walsh inhibitor of cAMP-dPK, whereas in some cases the stimulation of phosphorylation of the same proteins by exogeneous cAMP was relatively small. Phosphoproteins from two learning mutants known to be deficient in cAMP metabolism, dnc and rut, were analyzed for their extent of phosphorylation in the presence of a stable cAMP analogue; no significant differences from normal were detected, suggesting that the genetic defect in cAMP metabolism is not accompanied by constituent abnormalities in phosphorylated substrates in the adult fly, and that the physiological defects in these mutants result from aberrations in the interaction of the cAMP cascade with normal substrates. The majority of Ca2+/calmodulin kinase activity (80-90%, depending on the homogenization procedure) was associated with S2, as revealed by phosphorylation of exogenous synapsin I. Two endogenous substrates for this kinase in P2 had molecular masses of approximately 45 and 87 kilodaltons. At least 11 substrates for the Ca2+/calmodulin-dependent kinase were detected in S2.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3040908 TI - Solubilization of kainic acid binding sites from rat brain. AB - Kainic acid binding sites were solubilized from rat brain using a combination of Triton X-100 and digitonin. The highest percentage of solubilized binding sites (45%) was obtained by treating brain membranes with 1% Triton-X-100 and 0.2% digitonin in 0.5 M potassium phosphate containing 20% glycerol. The solubilized binding sites were stable and amenable to analysis by gel filtration and lectin affinity chromatography. Computer assisted analyses demonstrated that the solubilized sites displayed high- and low-affinity binding constants similar to the membrane-bound sites. Competition experiments further supported the pharmacological similarities of the solubilized and membrane-bound sites. Gel filtration chromatography of the solubilized binding site indicated that the detergent-bound complex had a Stokes radius of 82.7 A. The [3H]kainic acid binding site appears to be glycosylated based on its capability to bind to lectins. The lectin, wheatgerm agglutinin, proved to be a potentially useful tool for characterization because the solubilized binding sites were bound and eluted in relatively high yield. PMID- 3040909 TI - Mechanism of kainate toxicity to cerebellar neurons in vitro is analogous to reperfusion tissue injury. AB - The neuroexcitotoxin kainate has been used as a selective lesioning agent to model the etiology of a number of neurodegenerative disorders. Although excitotoxins cause susceptible neurons to undergo prolonged or repeated depolarization, the proximate metabolic pathology responsible for neuronal necrosis has remained elusive. We report here that kainate-induced death of cerebellar neurons in culture is prevented by inhibiting the enzyme xanthine oxidase, a cellular source of cytotoxic superoxide radicals (O2-.). Moreover, neurons are also protected from excitotoxin-induced death by the addition to the culture medium of either superoxide dismutase or mannitol, which scavenge superoxide and hydroxyl radicals, respectively, or serine protease inhibitor, which forestalls formation of xanthine oxidase. These findings indicate that excitotoxin-induced neuronal degeneration is mediated by superoxide radicals generated by xanthine oxidase, a mechanism partially analogous to that proposed for tissue damage seen upon reperfusion of ischemic tissues. PMID- 3040910 TI - Characteristics of partially purified nerve growth factor receptor. AB - Receptors for the nerve growth factor protein (NGF) have been isolated from three cell types [embryonic chicken sensory neurons (dorsal root sensory ganglia; DRG), rat pheochromocytoma (PC12) and human neuroblastoma (LAN-1) cells] and have been shown to be similar with respect to equilibrium dissociation constants. The present results demonstrate that there are multiple molecular weight species for NGF receptors from DRG neurons and PC12 cells. NGF receptors can be isolated from DRG as four different molecular species of 228, 187, 125, and 112 kilodaltons, and PC12 cells as three molecular species of 203, 118, and 107 kilodaltons. The NGF receptors isolated from DRG show different pH-binding profiles for high- and low-affinity binding. High-affinity binding displays a bell-shaped pH profile with maximum binding between pH 7.0 and 7.9, whereas low-affinity binding is constant between pH 5.0 and 9.1, with a twofold greater binding at pH 3.6. At 22 degrees C, the association rate constant was found to be 9.5 +/- 1.0 X 10(6) M-1 s-1. Two dissociation rate constants were observed. The fast dissociating receptor has a dissociation rate constant of 3.0 +/- 1.5 X 10(-2) s-1, whereas the slow dissociating receptor constant was 2.4 +/- 1.0 X 10(-4) s-1. The equilibrium dissociation constants calculated from the ratio of dissociation to association rate constants are 2.5 X 109-11) M for the high-affinity receptor (type I) and 3.2 X 10(-9) M for the low-affinity receptor (type II). These values are the same as those determined by equilibrium experiments on the isolated receptors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3040911 TI - Inactivation and reactivation of the multifunctional calmodulin-dependent protein kinase from brain by autophosphorylation and dephosphorylation: involvement of protein phosphatases from brain. AB - The multifunctional calmodulin-dependent protein kinase (calmodulin-kinase) from rat brain was autophosphorylated in a Ca2+- and calmodulin-dependent manner. The activity of the autophosphorylated enzyme was independent of Ca2+ and calmodulin. Calmodulin-kinase was dephosphorylated by protein phosphatase C from bovine brain, which is the catalytic subunits of protein phosphatases 1 and 2A. The holoenzyme of protein phosphatase 2A was also involved in the dephosphorylation of the enzyme. The autophosphorylated sites of calmodulin-kinase were universally dephosphorylated by protein phosphatase C. Calmodulin-kinase was inactivated and reactivated by autophosphorylation and dephosphorylation, respectively. Furthermore, the regulation of calmodulin-kinase by autophosphorylation and dephosphorylation was observed using calmodulin-kinase from canine heart. These results suggest that the activity of calmodulin-kinase is regulated by autophosphorylation and dephosphorylation, and that the regulation is the universal phenomenon for many other calmodulin-kinases in various tissues. PMID- 3040912 TI - Stimulation of inositol phosphate production by neurotensin in neuroblastoma N1E115 cells: implication of GTP-binding proteins and relationship with the cyclic GMP response. AB - The association of neurotensin to its receptor in differentiated neuroblastoma N1E115 cells led to a fast and transitory increase of the intracellular concentration in inositol triphosphate and inositol biphosphate, followed by a slower and more stable increase inositol monophosphate. The action of inositol 1,4,5-triphosphate on digitonin-permeabilized N1E115 cells resulted in a stimulation of cyclic GMP levels that mimicked that induced by neurotensin. Therefore, the cyclic GMP stimulation is probably a consequence of the initial inositol triphosphate formation triggered by neurotensin. Fluoroaluminate ions and pertussis toxin had the capacity to modulate positively and negatively, respectively, the formation of inositol triphosphate induced by neurotensin, indicating that GTP-binding proteins are involved in the regulation of inositol phosphate levels by neurotensin receptors. PMID- 3040914 TI - Chronic relapsing inflammatory polyneuropathy complicating sicca syndrome. PMID- 3040913 TI - Generalised peripheral nerve dysfunction in acromegaly: a study by conventional and novel neurophysiological techniques. AB - Twenty four patients with clinical, radiological and biochemical evidence of acromegaly were investigated by a number of independent neurophysiological tests. Two-thirds of the patients showed evidence of generalised peripheral nerve dysfunction. A significant correlation was found between total exchangeable body sodium, an indicator of disease activity, and the severity of the neuropathy. The generalised peripheral nerve abnormality was found to occur independently of the associated carbohydrate intolerance human growth hormone levels and other endocrinological dysfunction in this disorder. PMID- 3040915 TI - Peripheral neuropathy of dietary riboflavin deficiency in chickens. AB - A strain of rapidly growing meat-type chickens was fed a diet deficient in riboflavin from 1-40 days of age. Diminished growth rate, progressive gait abnormality and reluctance to move were noted beginning on day 8. Neurologic abnormalities were related to peripheral neuropathy characterized by Schwann cell hypertrophy and degeneration with cytoplasmic lipid droplets' and segmental demyelination. Lesions were initially detected on day 10, and in concert with clinical signs became more profound between days 14 and 21. Sequestration of myelin debris within Schwann cells was common. Other features of the neuropathy included the presence of endoneurial edema and axonal degeneration involving small numbers of fibers. Remyelination of peripheral nerve fibers in birds on the deficient diet was occasionally seen on day 10, became progressively more prominent, and was marked by day 37. There was an associated, variable but incomplete, clinical improvement evident in later stages of the study. Liver concentrations of riboflavin in deficient birds were significantly reduced on day 13 but not on day 26. This neuropathy may be related to diminished tissue levels of the riboflavin-based coenzymes flavin-adenine dinucleotide (FAD) and flavin mononucleotide (FMN) leading to reduced cellular energy levels and profoundly affecting Schwann cells at some critical point in growth. PMID- 3040916 TI - Diagnosis of progressive multifocal leukoencephalopathy by brain biopsy with biotin labeled DNA:DNA in situ hybridization. AB - DNA:DNA in situ hybridization using a cloned JC virus (JCV) DNA probe labeled with biotin confirmed the presence of JCV DNA in formalin-fixed, paraffin embedded brain biopsies from four cases of progressive multifocal leukoencephalopathy (PML). Only small pieces of tissue were available in each case. Detection of the JC DNA:DNA hybrids was carried out by affinity cytochemistry. JCV DNA was identified predominantly in the nuclei of interfascicular oligodendrocytes in demyelinated areas of the biopsies. JC virus was isolated from one case, and the diagnosis of PML was substantiated in all cases by electron microscopic identification or immunocytochemical labeling of JC viral antigen. In situ hybridization using a biotin labeled JCV DNA probe is a specific, sensitive and convenient method for confirming the diagnosis of PML in suspected cases evaluated by brain biopsy. PMID- 3040917 TI - The distribution of (Na+ + K+)ATPase is continuous along the axolemma of unensheathed axons from spinal roots of 'dystrophic' mice. AB - (Na+ + K+)ATPase-like immunoreactivity along the axolemma of sensory and motor neurons and the plasmalemma of Schwann cells from spinal roots of dystrophic mice (129 ReJ Dy/Dy) was determined using polyclonal antibodies specific for guinea pig renal (Na+ + K+)ATPase (GP-17), along with polyclonal (439-2) and monoclonal (9A5) antibodies specific for rat renal (Na+ + K+)ATPase. In normal and dystrophic mice, (Na+ + K+)ATPase-like immunoreactivity was observed along the axolemma at nodes of Ranvier using GP-17 and 439-2, each of which binds to isozymes of (Na+ + K+)ATPase composed of the alpha and alpha + forms of the catalytic subunit. Staining was not seen along the nodal axolemma with 9A5, a preparation that binds to the alpha form of the catalytic subunit. The terminal processes and microvilli of Schwann cells were stained using all three antibody probes. The axolemma of unensheathed axons in dystrophic mice was continuously and uniformly labelled with GP-17 and 439-2, but not 9A5. Concentrations of (Na+ + K+)ATPase-like immunoreactivity along Schwann cell processes were observed most often in areas adjacent to unensheathed axolemma. At heminodes, staining abruptly decreased along Schwann cell processes in areas that were separated from the unensheathed axolemma by other intervening Schwann cell processes. It was concluded from these data that in dystrophic mice (Na+ + K+)ATPase is uniformly distributed along unensheathed portions of axons without evidence of detectable focal concentrations of the enzyme, and that the catalytic subunit of (Na+ + K+)ATPase along unensheathed axons is distinct from the alpha form found in Schwann cells and other organs. In addition, (Na+ + K+)ATPase is concentrated along the plasmalemma of Schwann cells in regions of close apposition to axolemmal areas associated with large ionic fluxes. PMID- 3040918 TI - Chemotherapy for adenocarcinoma of the lung (WHO III): A randomized study of vindesine versus lomustine, cyclophosphamide, and methotrexate versus all four drugs. AB - Two hundred seventy-nine patients with previously untreated nonresectable adenocarcinoma of the lung (ACL) entered a prospective randomized trial, comparing vindesine (VDS) to a combination of lomustine (CCNU), cyclophosphamide (CTX), and methotrexate (MTX), and to a regimen including all four drugs. Response assessment was possible in 218 patients, while 259 were evaluable for survival. Response rates were similar (22%, 23%, and 27%, respectively) as were median durations of response (15 weeks overall) and survival (29 weeks overall). Patients with dose-limiting toxicity had significantly higher response rate and longer survival than patients without toxicity. The major toxicity was peripheral neuropathy with VDS treatment and myelosuppression with the other two regimens. The VDS single-agent activity in ACL was confirmed, but addition of VDS to the three-drug regimen did not increase activity. Future studies of VDS in combination with other active agents, and comparison to a matched control group on supportive care, are indicated. PMID- 3040919 TI - Eight drugs in one day chemotherapy for brain tumors: experience in 107 children and rationale for preradiation chemotherapy. AB - The development of a new multidrug chemotherapy regimen for primary brain tumors was based upon the cellular heterogeneity within individual tumors, the Goldie Coldman and Price-Goldie-Hill hypotheses, and known agonistic effects of certain drug combinations and sequences. Eight drugs (vincristine [VCR], hydroxyurea, procarbazine, CCNU, cisplatin, cytosine arabinoside [Ara-C] high-dose methylprednisolone, and either cyclophosphamide or dacarbazine) were administered within 12 hours in an attempt to minimize myelosuppression. Courses were repeated at 2- to 4-week intervals. The regimen was devised to include lipid and water soluble drugs, polar and nonpolar agents, phase-specific and cell-cycle independent agents, and antineoplastics with different mechanisms of action. More than 330 courses of the regimen were administered to 107 children with brain tumors whose tumor had recurred or had been incompletely resected at diagnosis. Tumor response according to protocol-specified criteria and independent review was evaluable in 78% of the patients. After just two cycles of chemotherapy and within a 4- to 6-week interval, 50% had an objective tumor response including 15.5% who had a complete response (CR). Nephrotoxicity and high-frequency hearing losses were noted after three to five courses of therapy in approximately half of the patients. Transfusions with red cells or platelets and use of antibiotics for fever and neutropenia were required in 10% to 25% of patients. The regimen appears satisfactory for preradiation chemotherapy in newly diagnosed patients with residual tumor after surgery, but it must be compared with standard therapeutic approaches in prospective controlled trials before its relative value can be established. PMID- 3040921 TI - Klinefelter's syndrome associated with mediastinal germ cell neoplasms. AB - Several case reports have suggested an association of primary mediastinal germ cell tumor (PMGCT) and Klinefelter's syndrome (KS). In an effort to confirm this association, 22 patients with mediastinal germ cell tumors had chromosome studies performed in a prospective fashion. Five patients (22%) had karyotypic or pathologic evidence of KS. All of the patients with KS had germ cell tumors of the nonseminomatous subtype and were relatively young (median age, 15 years). The literature confirms the findings of a young median age (18 years), nonseminomatous subtype, and mediastinal location of the germ cell neoplasm. We conclude that patients with KS are predisposed to the development of mediastinal nonseminomatous germ cell cancers. PMID- 3040920 TI - Platinum analogue combination chemotherapy: cisplatin and carboplatin--a phase I trial with pharmacokinetic assessment of the effect of cisplatin administration on carboplatin excretion. AB - Cisplatin (NSC 119875) and carboplatin (NSC 241240) are platinum (II) analogues with very different spectra of toxicity. Cisplatin dose is limited by nausea and vomiting, renal dysfunction, and dose-related peripheral neuropathy, whereas carboplatin is myelosuppressive. There are also clinical and laboratory data that suggest that these drugs may not be completely cross-resistant. Therefore, the following phase I trial of combination therapy with cisplatin and carboplatin was undertaken. Since carboplatin toxicity is enhanced in the presence of renal impairment, carboplatin excretion was also evaluated in selected patients at the maximum tolerated dose. Thirty-three patients received 50 mg/m2 cisplatin and doses of carboplatin between 160 mg/m2 and 400 mg/m2. Sequential 20-minute infusions of carboplatin and then cisplatin were able to be administered at the standard doses of carboplatin (320 and 400 mg/m2) with thrombocytopenia to the degree expected if carboplatin alone had been given. However, 280 mg/m2 carboplatin followed by 25 mg/m2 cisplatin/d X 3 caused unexpectedly severe thrombocytopenia in seven of eight patients (median platelet nadir 45,000/microL; range, 12 to 321,000/microL; nadir was less than 90,000 in seven of eight patients). In three patients treated with 280 mg/m2 carboplatin plus 25 mg/m2/d X 3 cisplatin, pharmacokinetics of carboplatin were compared during consecutive monthly cycles without and with cisplatin. Modestly increased areas under the curve (AUC) for carboplatin (15% and 35%) with cisplatin were seen in the two patients who experienced more pronounced platelet suppression with combination therapy. No other limiting or unusual toxicity was seen with this combination. Responses, primarily in "platinum responsive" tumors, were seen. The combination of cisplatin plus carboplatin is feasible and merits further study. PMID- 3040922 TI - Effects of patient management guidelines on physician practice patterns: the Community Hospital Oncology Program experience. PMID- 3040923 TI - Canadian multicenter randomized trial comparing sequential and alternating administration of two non-cross-resistant chemotherapy combinations in patients with limited small-cell carcinoma of the lung. AB - In order to assess the effect of scheduling of chemotherapy on the outcome of patients with limited small-cell lung cancer (SCLC), the Clinical Trials Group of the National Cancer Institute of Canada carried out a randomized trial comparing the alternation of cyclophosphamide, Adriamycin (Adria Laboratories, Columbus, OH; doxorubicin) and vincristine (CAV) with etoposide (VP-16) and cisplatin for six cycles to the administration of these two combinations in a sequential fashion (three cycles of CAV followed by three of VP-16/cisplatin). Three hundred eligible patients were enrolled on the trial from September 1981 to October 1984. All responding patients were also treated after completion of chemotherapy with thoracic irradiation in randomly allocated doses of 2,000 and 3,750 cGy. The complete response (CR) rate to chemotherapy was slightly, but not significantly, higher on the alternating arm (52% v 44%, P = .20). However, there was no difference in disease-free or overall survival on the alternating and sequential arms, respectively (47.3 weeks v 45.1 weeks, P = .26; 61.7 weeks v 59.5 weeks, P = .56). Data on the effect of radiotherapy dose on survival are not yet mature, but it does not appear the results of this portion of the trial will alter the interpretation of the chemotherapy comparison. Patient characteristics favorably influencing survival were female sex, good performance status, younger age, and absence of supraclavicular node involvement. Two interpretations of these and other results in SCLC are suggested: (1) the difference between the schedules used is too small for the predictions of the Goldie-Coldman model to be realized in a trial of this size, or (2) VP-16/cisplatin is actually a superior regimen and any schedule that exposes patients to these drugs early in treatment will produce improved results. PMID- 3040924 TI - Molecular cloning of a 2',3'-cyclic nucleotide 3'-phosphodiesterase: mRNAs with different 5' ends encode the same set of proteins in nervous and lymphoid tissues. AB - Antibodies raised to a mixture of the 46 and 48 kDa rat CNS 2',3'-cyclic nucleotide 3-phosphodiesterases (CNPs) recognized apparently identical proteins in peripheral nervous system (PNS), thymus, and circulating blood lymphocytes. These antibodies were used to identify, in a rat brain phage lambda gt11 expression library, cDNA clones encoding beta-galactosidase-CNP fusion proteins, some of which showed CNP activity. In RNA blots, the subcloned CNP cDNA inserts hybridized to mRNAs of approximately 2400 and approximately 2800 nucleotides (nts), and to a approximately 2500 nt mRNA from thymus. Several nonexpressing CNP cDNAs were identified by plaque hybridization, and the mRNA transcribed in vitro from one of these cDNAs (pCNP7) encoded a complete 46 kDa CNP polypeptide. Examination of the deduced amino acid sequence revealed an apparent homology to cAMP binding sites in several other proteins. A 373 bp segment from the 5' end of this pCNP7 hybridized only to the 2800 nt nervous system mRNAs, thus revealing that not all CNP mRNAs share the same 5'-ends. Genomic DNA blots probed with CNP cDNAs suggest that there is a single gene which can be alternatively spliced to produce the various mRNA transcripts in the nervous and lymphoid tissues. PMID- 3040925 TI - N-methyl-D-aspartate receptor-induced, inherent oscillatory activity in neurons active during fictive locomotion in the lamprey. AB - Bath application of N-methyl-aspartate induces fictive locomotor activity in the isolated spinal cord preparation of the lamprey, as well as TTX-resistant membrane potential oscillations in many individual neurons. This inherent oscillatory activity is shown to depend on a specific activation of N-methyl-D aspartate (NMDA) receptors. This activation initiates voltage-dependent, magnesium-requiring membrane potential bistability, presumably due to a development of a region of negative slope conductance in the current-voltage relation of the neuron. When sodium ions were removed from the bathing solution, oscillations disappeared, and the membrane potential was maintained at a hyperpolarized level, suggesting that the depolarizing current during the oscillatory cycle is mainly carried by sodium ions. Replacing Ca2+ with Ba2+ also leads to a cessation of oscillatory activity, with the membrane potential remaining at the more depolarized level. This indicates an involvement of a Ca2+ dependent K+ current during the repolarization phase. These findings, together with the voltage dependence, can account for the main characteristics of the NMDA receptor-induced, TTX-resistant membrane potential oscillations. This oscillatory behavior has been demonstrated in motoneurons and in several interneurons including CC interneurons but has not been found in edge cells, dorsal cells, or lateral interneurons. The possibility that inherent oscillatory membrane properties may contribute to the activity pattern during fictive locomotion was investigated in experiments with intracellular current injection in the absence of TTX. The stimulation effects obtained required the presence of magnesium ions and were analogous to the stimulation effects seen during oscillations after TTX blockade. Together with similarities in, for instance, frequency and amplitude between the locomotor oscillatory activity and the TTX-resistant oscillations, the results are compatible with an involvement of inherent, oscillatory membrane properties during fictive locomotion in the lamprey spinal cord. PMID- 3040926 TI - Hypophysectomy increases vasoactive intestinal peptide-stimulated cyclic AMP generation in the hippocampus of the rat. AB - In investigating the feedback effects of circulating hormones on the brain, we showed previously that adrenalectomy (ADX) increases vasoactive intestinal peptide (VIP)-stimulated cAMP generation in slices from rat hippocampus, a brain structure with high levels of glucocorticoid receptors. This effect is reversed by replacement with glucocorticoids such as dexamethasone (DEX) and corticosterone (CORT). Here we report that, like ADX, hypophysectomy (HYPOX) also elevates VIP-stimulated cAMP generation, compared with sham-operated controls. Moreover, like glucocorticoid replacement, administration of ACTH to HYPOX rats causes a decrease in cAMP stimulation by VIP. Furthermore, ACTH had no effect when given to HYPOX + ADX rats, indicating that the effects of ACTH require the presence of adrenal steroid secretion. However, we find that ACTH may have a permissive role in this glucocorticoid effect because, in the absence of the pituitary, DEX treatment does not decrease VIP-stimulated cAMP levels in the hippocampus. In addition, hippocampal beta-adrenergic-stimulated cAMP accumulation was not suppressed by DEX treatment of HYPOX rats, which again is different from the effect of DEX treatment on ADX animals. These results are discussed in terms of possible synergism between pituitary hormones and steroid hormones in exerting feedback actions on brain function. PMID- 3040927 TI - Octopamine- and cyclic AMP-stimulated phosphorylation of a protein in Limulus ventral and lateral eyes. AB - The biogenic amine octopamine (OCT) fulfills most of the criteria as a neurotransmitter of efferent fibers that project to lateral and ventral eyes of the horseshoe crab, Limulus polyphemus. OCT is synthesized by and released from the efferent fibers, and OCT mimics many of the effects of endogenous efferent activity. OCT stimulates an increase in intracellular adenosine 3',5' monophosphate (cAMP) in both ventral and lateral eyes, and many of the physiological effects of OCT in these eyes appear to be mediated via cAMP dependent mechanisms. Here we show that OCT, acting apparently through an OCT specific receptor, stimulates the increased phosphorylation of a protein with an apparent molecular weight of 122 kDa in both ventral and lateral eyes. This protein is also phosphorylated in response to 8-bromo cAMP and forskolin, suggesting that its phosphorylation involves activation of a cAMP-dependent protein kinase. We present evidence that the 122 kDa protein may be widely distributed in the Limulus visual system but that its phosphorylation in intact tissue in response to OCT, or agents acting through cAMP, may be restricted to portions containing photoreceptor cell bodies. The 122 kDa protein is quantitatively a major cellular protein in the photoreceptor cell body enriched portions of the ventral eye, its isoelectric point is between pH 6.2 and 6.4, and it is associated with both cell membranes and the cytoplasm. The function of this protein is not yet known. It may be important in mediating one or more of the effects of octopamine on Limulus vision. PMID- 3040928 TI - Effects of an anxiogenic benzodiazepine receptor ligand on motor activity and dopamine release in nucleus accumbens and striatum in the rat. AB - The effects of the anxiogenic beta-carboline FG 7142 (N-methyl-beta-carboline-3 carboxylate) on motor activity and dopamine release in nucleus accumbens and striatum were measured in the rat. Changes in extracellular homovanillic acid (HVA) concentration, monitored by computer-controlled linear sweep voltammetry with carbon-paste electrodes, were used as an index of changes in dopamine release. An intraperitoneal injection of FG 7142 was followed by an inhibition of the nocturnal rise in motor activity and in dopamine release in nucleus accumbens, but not striatum. Two days after the drug injection, dopamine release in nucleus accumbens returned to control level and then increased on days 3-6 after the injection; there was no delayed change in motor activity or in striatal dopamine release. In parallel experiments using ex vivo changes in the ratio of 3,4-dihydroxyphenylacetic acid (DOPAC) to dopamine as an index of changes in dopamine turnover, a similar early depression and delayed increase of dopamine turnover in nucleus accumbens, with no change in striatum, was found after an intraperitoneal injection of FG 7142. Regression analysis of motor activity versus dopamine release showed a decrease in correlation between these 2 parameters for nucleus accumbens but not striatum after FG 7142 injection. These results suggest that the inverse benzodiazepine receptor agonist FG 7142 has a biphasic effect on dopamine release from mesolimbic neurons and support the hypothesis that dopamine release in nucleus accumbens and striatum has a modulatory effect on the control of motor activity but does not play a determining role in the regulation of movement. PMID- 3040929 TI - Diagnosis of pericardial effusions from routine gated blood-pool imaging. AB - Gated blood-pool scintigraphy (GBPS) is often obtained as the initial test to evaluate symptoms suggestive of left ventricular dysfunction. Since large pericardial effusions may also cause such symptoms, the ability to recognize them on routine GBPS is of clinical importance. Characteristic features of the "halo" sign surrounding the cardiac blood pool were developed, based on the GBPS of patients with known pericardial effusions. These criteria were then applied blindly to 154 consecutive patients who underwent both GBPS and echocardiography. All five patients with large effusions (approximately greater than 500 ml) were correctly identified by GBPS (sensitivity 100%); for patients with moderate effusions (approximately 150-500 ml), the sensitivity was only 33% (3/9). There were three false positives (specificity 98%). We conclude that large pericardial effusions can be identified with high sensitivity and specificity on routine GBPS. Although echocardiography remains the method of choice for the diagnosis of effusions, inspection for characteristics suggesting their presence on GBPS should be part of routine interpretations. PMID- 3040930 TI - Rival conceptions in doctoral education in nursing and their outcomes: an update. AB - The need for two distinct types of doctoral programs in nursing was historically well conceived and well articulated. The growing literature on the doctoral education in nursing, however, shows that the implementation of the concept has not produced the intended results, that is the coexistence of academic and professional doctoral programs in nursing has not resulted in the differentiated career paths for the graduates. Instead, the competing aims of doctoral education in nursing, paved the way to varying doctoral titles, and weakened the claims of professional doctoral programs as preparing for advanced clinical practice. More significantly, however, this development is threatening the future existence of professional doctoral programs in nursing. PMID- 3040931 TI - Role-taking skills and perceived similarity in baccalaureate nursing students. AB - The importance of exercising effective role-taking skills as a nursing care provider is described. Twenty-six baccalaureate nursing students provided role taking responses to four real life events. Students also reported whether or not they had ever experienced similar events and to what degree they perceived themselves as similar to the individuals in the events on a number of personal characteristics. Role taking was compared across the situations. Having experienced a similar life event or perceiving similarity to the main character did not appear to enhance role-taking responses. The results suggest that it may not be necessary to have had a life experience or background similar to that of the client's in order to be perceived as a nurse who is caring and understanding. Implications for nursing students are discussed. PMID- 3040932 TI - Perceived role conflict, role ambiguity, and job satisfaction among nurse educators. PMID- 3040933 TI - Health policy in nursing curriculum. AB - This article presents the findings of a study on how health policy is integrated into the nursing curriculum at both the graduate and undergraduate levels, with particular emphasis on type of coursework, faculty preparation, and teaching strategies. The sample for the study included 210 randomly selected schools of nursing: 150 with undergraduate programs only and 60 with both graduate and undergraduate programs. The findings indicate that health policy has not yet received optimal placement in the curriculum at either the undergraduate or graduate level of nursing preparation. Recommendations are made based upon the findings of the study, a review of the literature, and the collective experience of the authors. PMID- 3040934 TI - A wellness clinic for older persons: a positive experience for nursing students and older persons. PMID- 3040935 TI - Let's take the "i" out of failure. PMID- 3040936 TI - The clinical application of mental health concepts in the integrated curriculum. PMID- 3040937 TI - Preparation for NCLEX-RN: an innovative course. National Council Licensure Examination. AB - In summary, a course with a content review format is frustrating to both students and faculty because an attempt is made to cover all nursing clinical disciplines in a short time period. The preparation for NCLEX-RN course format was changed from a primarily content review to a test-taking approach. The student is given a Decision-Making Model for eliminating response choices for questions to which they do not already know the answer. This was found to be a positive change for both students and faculty. This innovative approach to the NCLEX-RN review would be beneficial for other state board review courses. PMID- 3040938 TI - Part-time status in university teaching: an ambiguous state. PMID- 3040939 TI - Occupational health nursing as a component of baccalaureate nursing education. AB - Much can be gained by the nursing profession and by society by having well prepared, professional occupational health nurses. Furthermore, conceptual knowledge about occupational health will better prepare the non-specialized nurse for her or his nursing practice. Lastly, knowledge about the hazards and risks that may accompany one's occupation will increase the nurse's awareness of the hazards that she/he may face in her/his own occupational setting. For these reasons the incorporation of occupational health content in the baccalaureate nursing programs would be a singular achievement for the nursing profession. It is clear that work-related injuries and illnesses pose a major challenge to today's health care providers. Most occupational health problems are preventable. It is fitting and indeed imperative that members of the nursing profession be leaders in a concerted effort to reduce the many risks in the workplace. The U.S. Public Health Service (1983) states that it is their goal that "by 1990 at least 70% of primary health care providers should routinely elicit occupational health exposures as part of the patient (health) history and should know how to interpret the information in an understandable manner." It is only through the introduction of important occupational health concepts in baccalaureate schools of education that this goal will be accomplished. Knowledge about occupational health nursing is fragmented. Furthermore, an occupational health curriculum, when it does exist, is varied in its content and its emphasis. this stems in part from a dearth of knowledge and experience by educators in the field of occupational health.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3040940 TI - Interdisciplinary health team care: nursing education in a rural health setting. PMID- 3040941 TI - Estimation of the potential digestibility and rate of degradation of water insoluble dietary fiber in the pig cecum with a modified nylon bag technique. AB - Potential digestibility and rate of degradation of water-insoluble material (WIM) prepared from rutabaga, wheat bran and digesta recovered from the terminal ileum of pigs fed bran- or rutabaga-based diets were measured in the cecum of surgically modified pigs. WIM samples recovered from the cecum after fixed incubation times were analyzed and the rate of total disappearance of organic matter, cellulose, uronic acid and noncellulosic neutral sugars was calculated. Maximum degradation of each WIM occurred within 48 h after incubation and was for rutabaga and its ileal digesta 90 and 70%, respectively, and for bran and bran ileal digesta 55 and 45%, respectively. Cellulose and uronic acid in bran samples did not appear to be degraded, material disappearance being attributable to the loss of noncellulosic neutral sugars. Estimation of potential organic matter digestibility showed rutabaga to be 93%, rutabaga digesta 79%, bran 56% and bran digesta 43% digestible. Fractional rates of digestion in rutabaga samples were significantly faster than in corresponding bran samples. When related to the role of fiber in nutrition the results obtained suggest that cereal fiber would be only slightly modified during gut transit, unlike vegetable fiber in which microbial digestion would probably result in the total destruction of the fiber cell wall matrix. PMID- 3040942 TI - The spectrum of cytomegalovirus infection following human heart-lung transplantation. AB - Data were analyzed from 19 long-term survivors of cardiopulmonary transplantation in this institution, including nine patients with normal pulmonary function and 10 recipients with posttransplant obliterative bronchiolitis. In all cases, donor cytomegalovirus titers (IgG), preoperative recipient titers (IgG), and serial postoperative recipient titers (IgM, IgG, and complement fixation) were available. In addition, surveillance cytomegalovirus cultures and pulmonary function tests were obtained prospectively after surgery in all 19 patients. A total of 12 patients developed active cytomegalovirus infection (serologic conversion confirmed by positive cultures) after transplantation, six of whom subsequently developed obliterative bronchiolitis. However, infection was clinically associated with pulmonary deterioration in only four of these patients, three of whom had cytomegalovirus pneumonitis. With the exception of obliterative bronchiolitis, no other permanent sequelae of cytomegalovirus infection were evident in this small group. Progressive obliterative bronchiolitis was also seen in four of the seven recipients who had no evidence of cytomegalovirus infection at any time. Although viral causes have been associated with obliterative bronchiolitis, the current data suggest that cytomegalovirus infection in the absence of pneumonitis does not appear to be a significant risk factor for obliterative bronchiolitis in cardiopulmonary transplant recipients. A larger group of patients will be required to ultimately establish the role of cytomegalovirus infection in this setting. PMID- 3040943 TI - Fatal cytomegalovirus infection and coronary arterial thromboses after heart transplantation: a case report. AB - A 36-year-old woman died because of disseminated cytomegalovirus infection after a heart allograft transplantation was performed. Autopsy revealed that widespread coronary arterial thromboses were associated with cytomegalovirus infection of endothelial cells. Its possible pathogenetic mechanism and significance were discussed. PMID- 3040944 TI - Successful treatment with trisodium phosphonoformate for primary cytomegalovirus infection after heart transplantation. AB - Primary cytomegalovirus infection in the patient who is receiving immunosuppression therapy is associated with a high morbidity and mortality. We report a patient who developed primary cytomegalovirus infection 37 days after heart transplantation with a rapidly deteriorating course. Treatment with the new antiviral drug trisodium phosphonoformate (Foscarnet-Astra) was initiated as a lifesaving measure with rapid, dramatic improvement in the patient's condition and subsequent recovery. PMID- 3040945 TI - Ridge augmentation using solid and porous hydroxylapatite particles with and without autogenous bone or plaster. AB - Edentulous areas of dog jaws were augmented with solid or porous particles of hydroxylapatite (HA) alone, or combined with either finely crushed autogenous bone or plaster of paris. At the end of the experiment (24 weeks), the augmented ridges were firm and stable and covered with healthy mucosa. The ridges augmented with only porous particles of HA demonstrated a greater amount of bone ingrowth compared with the solid, dense particles. The new bone formation occurred in those parts of the implants adjacent to the underlying alveolar bone. The addition of autogenous bone to the HA particles did not enhance bony deposition, and none of the autogenous bone chips survived for 24 weeks. The amount of new bone in the ridges augmented with plaster of paris and HA was similar to the other groups, and the plaster did not interfere with healing. There was evidence of resorption of the underlying cortical bone in many of the specimens. PMID- 3040946 TI - Saliva composition and caries development during protein deficiency and beta receptor stimulation or inhibition. AB - We tested the hypothesis that a low protein diet (5%) would change the conditions for synthesis and release of protein from the salivary secretory cells and increase caries development. In addition, we tested whether the simultaneous use of a beta-adrenoceptor agonist or antagonist had an additive effect. After an experimental period of nine weeks the animals fed the 5% protein diet had lower body weights, saliva secretion rates and total protein secreted per minute than a control group fed a 20% protein diet. There were greater numbers of cariogenic streptococci on the teeth and the caries scores were higher in the rats fed the low protein diet than in the rats in the control group. The beta-receptor agonist, isoproterenol, given to rats fed the 20% protein diet, caused a reduction in protein concentration and amylase activity in saliva and a slight increase in caries development. Propranolol had no effect on either saliva composition or caries development. PMID- 3040947 TI - Expression of oncogenes in human tumours with special reference to the head and neck region. AB - A major recent advance in cancer research has been in the field of oncogenes. Oncogenes are genes with a proven cancer association and which appear to be particularly implicated in cellular regulation and proliferation. The oncogenic potential of specific cellular genes has now been recognised and this has influenced current thinking concerning the initiation of carcinogenesis. The specific role of an oncogene is still incompletely understood but research with one particular oncogene (ras) has demonstrated that it can be involved in more than one stage of multi-step carcinogenesis. New techniques are being developed and evaluated to determine the expression of specific oncogenes in normal and neoplastic tissues, with a view to using them in future diagnostic immuno histopathological methods. This review describes the concept of oncogenes and discusses their role in the development of neoplasia. The results of the expression of various oncogenes in human malignancies with special reference to the head and neck regions are discussed. Finally, the future prospects of this research field are examined their and its possible implications in cancer therapy. PMID- 3040948 TI - Nucleic acid probes in the study of latent viral disease. AB - Despite the apparent complexities of vocabulary and techniques involved in nucleic acid hybridization, these methods should become important new weapons in the pathologist's armoury. The particular strengths lie in the analysis of genes whose protein products defy detection either because they are absent (due, for example, to cellular controls as found in viral latency), to a point mutation, or to gene deletion (as is often the case with cellular oncogenes). However, the mere application of this technology will not solve all diagnostic problems. One must be aware, particularly in applying in situ hybridization to a new system, of the artefactual binding of the probes, and suitable control and duplicate experiments should be performed. In addition, it is vital to verify the identity of the probes and the specificity of the reaction by filter hybridization. The latter procedure may seem unwieldy, but is, in fact, no more complex than the recommended procedures for immunohistochemistry, as discussed by Matthews (this symposium). Sadly, for both hybridization and immunological detection, operator inexperience has often prevented such checks of specificity and as a result many spurious results populate the scientific literature, for example a recent controversy over the detection of Human T cell Lymphotrophic Virus Type 1 in multiple sclerosis patients. With careful application, however, these techniques will permit the detection of viral molecules under conditions where traditional electron microscopy/histology have failed, and may reveal possible viral aetiologies for a range of hitherto non-viral diseases. PMID- 3040949 TI - The evaluation of a biphasic calcium phosphate ceramic for use in grafting long bone diaphyseal defects. AB - The effectiveness of a sintered hydroxyapatite-tricalcium phosphate (HA-TCP) ceramic in bridging large diaphyseal defects in the canine ulna was studied. One hundred percent morselized HA-TCP, a 50:50 mixture of morselized HA-TCP, and autogenous cancellous bone, and 100% autogenous cancellous bone were used to bridge 2.5-cm defects in the left ulnae of three groups of six dogs each. At 24 weeks the ulnae were explanted and studied by radiography, microradiography, mechanical testing, and histology. Pure HA/TCP was not osteoinductive, and four of six ulnae in this group progressed to a fibrous nonunion. The HA/TCP cancellous bone mixture and pure cancellous bone were approximately equal in effect, leading to good callus formation at 4 weeks and strong bony union by 24 weeks, with no evidence of bioincompatibility. Morselized HA/TCP promises to be useful as a graft extender when mixed with autogenous cancellous bone. PMID- 3040950 TI - Breast carcinoma with stromal multinucleated giant cells. PMID- 3040951 TI - Response of mouse lung to crocidolite asbestos. 2. Pulmonary fibrosis after long fibres. AB - To determine the cellular and fibrogenic responses of the lung to long asbestos fibres, mice were instilled intratracheally with 0.1 mg of a sample of long crocidolite fibres. Animals were killed at intervals to 20 weeks with 3H thymidine injected one h before death. Following bronchoalveolar lavage, an increase in polymorph neutrophils (PMN) and alveolar macrophages (AM) was found during the first week, accompanied by elevated glucosaminidase and alveolar protein levels. Although the PMN number dropped, some were always recovered by lavage to 20 weeks. Early multifocal necrosis of bronchiolar epithelium was followed by a large increase in labelling of epithelial cells and underlying fibroblasts. Epithelial overgrowth of luminal long fibres and inflammatory exudates was followed by giant cell and granuloma formation in the interstitium. After four weeks collagen levels were significantly increased and fibrosis was seen in these peribronchiolar locations. A few small fibres were observed in AM but no evidence of fibrosis was seen in alveolar walls. These findings suggest that injury to bronchial and bronchiolar epithelium allows long fibres to reach the interstitium where subsequent macrophage-fibroblast interactions result in a severe fibrotic reaction that resembles the bronchiolar component of human asbestosis. PMID- 3040952 TI - Demonstration of human papillomavirus types in paraffin processed tissue from human ano-genital lesions by in-situ DNA hybridisation. AB - A sensitive in situ hybridization technique for the demonstration of human papillomavirus (HPV) employing a biotin-streptavidin polyalkaline phosphatase complex has been successfully applied to formalin-fixed, paraffin processed tissue obtained from a selected series of patients with ano-genital lesions. Benign condylomata from males and females showed the presence of HPV 6 and 11. Two cases of vulval intraepithelial neoplasia showed HPV 16. Four cases of squamous carcinoma of the anal canal also showed HPV 16 in the tumour or in the adjacent pre-invasive neoplastic epithelium. A case of malignant transformation in a cervical condyloma was associated with HPV 6 and 11. This technique permits the retrospective evaluation of routinely processed material thus widening the investigative spectrum for HPV. PMID- 3040954 TI - Wilms tumor in three patients with Bloom syndrome. PMID- 3040953 TI - Response of mouse lung to crocidolite asbestos. 1. Minimal fibrotic reaction to short fibres. AB - To determine the relationship between the development of pulmonary fibrosis and the size of deposited asbestos, we prepared a pure sample of short crocidolite fibres and instilled 0.5 mg of 0.1 mg to the lungs of mice. Animals were killed up to 20 weeks later with 3H thymidine injected 1 h before death. By bronchoalveolar lavage, there was a rapid transient increase in polymorph neutrophils (PMN) and in glucosaminidase levels; alveolar macrophage (AM) numbers were elevated in the 0.5 mg group for eight weeks. Most fibres were phagocytized by AM, many of which were heavily laden and cleared from the lung over the 20 week period. Some fibres were seen in type 1 epithelial cells, frequently associated with cell injury. From cell kinetic studies, a very brief proliferative response was seen in bronchiolar epithelial and Type 2 alveolar epithelial cells. A greater response was seen in interstitial fibroblasts which showed increased labelling up to two weeks after 0.5 mg asbestos. However no granulomas were seen and very little fibrosis was found by morphology or by biochemistry at any time after 0.5 mg; no fibrosis was seen after instilling 0.1 mg. The results show that a high dose of exclusively short asbestos fibres produces minimal lung injury and fibrosis in spite of long standing macrophage fibre interaction in the alveoli. PMID- 3040955 TI - Reevaluation, using marker enzymes, of the ability of saponin and ammonium chloride to free Plasmodium from infected erythrocytes. AB - Saponin and ammonium chloride lysis have been applied for some time to the separation of erythrocyte membranes from malarial-infected erythrocytes, allowing easy isolation of the parasites. We present a reevaluation of the use of saponin and ammonium chloride as tools for isolating Plasmodium (knowlesi or falciparum) parasites. Acetylcholine esterase (EC 3.1.1.7) was used as an erythrocyte membrane marker and CDP-choline: 1,2-diacylglycerol cholinephosphotransferase (EC 2.7.8.2) as a parasite membrane marker to monitor fractionation by these agents. Both saponin and ammonium chloride produced hemolysis of uninfected and infected erythrocytes, but failed to separate host erythrocyte membrane from the parasite, regardless of its stage. Thus, saponin and ammonium chloride can be used to isolate whole infected erythrocytes, depleted of hemoglobin, by selective disruption of uninfected cells. PMID- 3040956 TI - Nature of collagenolytic enzyme and inhibitor activities in crevicular fluid from healthy and inflamed periodontal tissues of beagle dogs. PMID- 3040957 TI - The biocompatibility of hydroxyapatite implanted in the human periodontium. PMID- 3040958 TI - Collagenolytic activity of black-pigmented Bacteroides species. PMID- 3040959 TI - Clinical evaluation of porous and nonporous hydroxyapatite in the treatment of human periodontal bony defects. AB - The purpose of this study was to investigate the effectiveness of a synthetic nonporous hydroxyapatite graft material (OrthoMatrix HA-500), a porous replamineform hydroxyapatite graft material (Interpore 200), and a debrided control with respect to defect fill. Twelve adult patients having periodontitis and three similar angular osseous defects as verified by radiographic analysis and clinical probe depths greater than or equal to 5 mm were selected. Clinical parameters gathered prior to surgical intervention and at identified postoperative visits included plaque index, probing depth, and standardized radiographic examination. Customized acrylic stents were used as fixed reference guides for the insertion of endodontic silver points to the base of each defect during both initial and 6-month reentry surgeries. A total of 36 defects were randomly assigned to one of three treatment modalities such that 12 defects received OrthoMatrix HA-500, 12 received Interpore 200, and 12 served as debrided controls. Comparison of nonporous hydroxyapatite, porous replamineform hydroxyapatite, and debrided control treatment modalities revealed a statistically significant improvement (P less than 0.05) in the mean per cent of defect resolved in only those sites treated with nonporous hydroxyapatite. Similar positive trends were seen in the sites treated with nonporous hydroxyapatite for mean reentry defect depth and mean defect fill although these were not statistically significant (P greater than 0.05). No other significant differences were noted. PMID- 3040960 TI - In vitro effect of cinnamic aldehyde, a main component of Cinnamomi Cortex, on human platelet aggregation and arachidonic acid metabolism. AB - The in vitro effect of cinnamic aldehyde, a main component of Cinnamomi Cortex, on platelet aggregation and arachidonic acid (AA) metabolism in human platelets was studied. Cinnamic aldehyde reduced platelet aggregation of both platelet rich plasma and washed platelets, dose-dependently. This compound also decreased the formation of the metabolites of AA such as thromboxane B2 (TXB2), 12-hydroxy heptadecatrienoic acid and 12-hydroxyeicosatetraenoic acid in collagen-stimulated washed platelets. The conversion of exogenous [14C]AA to cyclooxygenase metabolites or 12-lipoxygenase metabolite was not altered significantly by the addition of cinnamic aldehyde. On the other hand, collagen-induced release of [14C]AA and its metabolites from washed platelets prelabeled with [14C]AA was markedly reduced by the addition of cinnamic aldehyde. These results suggested that cinnamic aldehyde suppressed the release of AA from platelet membrane phospholipids and then reduced the formation of thromboxane A2. This inhibitory effect of cinnamic aldehyde on AA release and TXB2 formation may contribute to reduced platelet aggregation. PMID- 3040961 TI - Effect of nourseothricin (streptothricin) on the outer membrane of sensitive and resistant Escherichia coli strains. AB - Nourseothricin (streptothricin) causes disturbances (perforations) in the outer membrane of sensitive E. coli strains allowing lysozyme and deoxycholate, but not the periplasmic alkaline phosphatase to penetrate. EDTA slightly increases, but Mg++ ions slightly decrease this effect. The cell walls of three from four nourseothricin-resistant strains do not become permeable under these conditions, but remain sensitive against TRIS/EDTA. Nourseothricin is supposed to pass the outer membrane of sensitive bacteria via some kind of "self-promoting" pathway. This way can (but need not) be blocked in resistant strains. PMID- 3040962 TI - Regulation of intracellular pH in reticulospinal neurones of the lamprey, Petromyzon marinus. AB - 1. The regulation of intracellular pH (pHi) in lamprey reticulospinal neurones was investigated with pH-sensitive micro-electrodes based on a neutral carrier liquid membrane. Experiments were performed using an in vitro brain-stem preparation. 2. In HEPES-buffered solutions, extracellular pH (pHo) was consistently more acidic than the pH of the bathing solution (pHb). In HCO3(-) buffered solutions, the brain was also relatively acidic, but the brain pH gradient was smaller. 3. In HEPES- and HCO3(-)-buffered solutions, mean pHi was 7.40-7.50. This range was too high to be explained by a passive distribution of H+, OH- or HCO3-. 4. In nominally HCO3(-)-free, HEPES-buffered solution, cells were acid loaded by addition and subsequent withdrawal of NH4+ from the superfusate. pHi recovered from acid loading by an energy-dependent process in 10 20 min. Recovery from acid loading in HEPES-buffered solutions was blocked by exposure to amiloride. 5. Removal of extracellular Na+ caused a slow, accelerating fall of pHi. Return of Na+ to the bath caused an immediate reversal of this acidification, followed by a slow recovery of pHi. Measurement with Na+ sensitive micro-electrodes during acid loading showed a rapid rise in the intracellular Na+ activity [( Na+]i). 6. Following acid loading, transition from HEPES- to HCO3(-)-buffered solutions caused an increase in the acid extrusion rate of at least 48%. The effect of these solution changes was dependent on pHo. After blocking pHi recovery with amiloride, transition from HEPES- to HCO3(-) buffered Ringer plus amiloride produced a slow recovery of pHi. 7. Recovery from acid loading in HCO3(-)-buffered solutions was inhibited 65% by the anion transport blocker DIDS (4,4'-diisothiocyanostilbene-2,2'-disulphonic acid). Recovery from acid loading after incubation in Cl(-)-free solution was slower than recovery after replenishment of Cl-. 8. It is concluded that in HCO3(-)-free solutions, pHi regulation is accomplished by a Na-H exchange mechanism. In the presence of extracellular HCO3- an additional mechanism can operate to extrude intracellular acid. PMID- 3040963 TI - Organization of synaptic transmission in the mammalian solitary complex, studied in vitro. AB - 1. Synaptic transmission and neuronal morphology were studied in the nucleus tractus solitarius and in the dorsal vagal motor nucleus (solitary complex), in coronal brain-stem slices of rat or cat, superfused in vitro. 2. Electrical stimulation of afferent fibres of the solitary tract evoked two different types of post-synaptic response recorded intracellularly in different solitary complex neurones. Labelling with horseradish peroxidase showed that these two sorts of orthodromically evoked responses were correlated with different post-synaptic neuronal morphologies. 3. The majority of recorded neurones (n = 93) showed a prolonged reduction in excitability following the initial solitary-tract-evoked excitatory post-synaptic potential (e.p.s.p.). A smaller number of neurones (n = 53) showed a prolonged increase in excitability following solitary tract stimulation. In no case did the solitary tract stimulation induce a burst of action potentials at high frequency. 4. The time-to-peak and the half-width of the initial solitary-tract-evoked e.p.s.p. were shorter in neurones with prolonged increased excitability than in those with prolonged reduced excitability. In neurones with prolonged reduced excitability, this e.p.s.p. was followed by a hyperpolarization lasting 60-100 ms. The latency of this inhibitory post-synaptic potential (i.p.s.p.) was 3-5 ms longer than that of the initial e.p.s.p. and its reversal potential was 10 mV more negative than the reversal potential of the response measured following application of gamma-aminobutyric acid or glycine. In neurones with prolonged increased excitability, at a membrane potential of -40 to -50 mV, the initial solitary tract e.p.s.p. was followed by a prolonged depolarization lasting 100-400 ms. 5. Background synaptic activity was high in neurones with prolonged increased excitability, consisting of unitary e.p.s.p.s with an amplitude of more than 0.8 mV. This activity was increased for a period of 300-800 ms following solitary tract stimulation. Spontaneous excitatory potentials of more than 0.5 mV were not seen in neurones with prolonged reduced excitability. In these neurones, after intracellular injection of choride ions, reversed unitary i.p.s.p.s formed a background activity which was increased following stimulation of the solitary tract. 6. Neurones with prolonged reduced excitability were found in the medial, ventral and ventrolateral part of the nucleus tractus solitarius and in the dorsal vagal motor nucleus where they were identified by their antidromic response to stimulation ventral and lateral to the tractus solitarius.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3040964 TI - Cyclic adenosine monophosphate differently affects the response of mouse pancreatic beta-cells to various amino acids. AB - 1. The membrane potential of mouse beta-cells was measured in parallel with 86Rb+ efflux and insulin release from mouse islets during stimulation by three types of amino acids and modulation of their effects by glucose and cyclic adenosine monophosphate (cyclic AMP) (forskolin being used to activate the adenylate cyclase). 2. In the absence of glucose, alanine and arginine accelerated 86Rb+ efflux, whereas leucine decreased it. They all depolarized the beta-cell membrane and slightly increased insulin release. Forskolin had little effect on 86Rb+ efflux, consistently potentiated insulin release but induced electrical activity only in the presence of leucine. 3. The effects of the three amino acids on 86Rb+ efflux and beta-cell membrane potential were not qualitatively altered by a non stimulatory concentration of glucose (3 mM). However, the release of insulin induced by leucine alone or with forskolin was markedly amplified, in contrast to that of alanine or arginine, which was inhibited. 4. In the presence of a threshold concentration of glucose (7 mM), the three amino acids accelerated 86Rb+ efflux and depolarized the beta-cell membrane. With alanine and arginine, spike activity was transiently observed and coincided with a short-lived increase in insulin release. With leucine, slow waves with superimposed bursts of spikes occurred and were accompanied by a sustained release of insulin. Forskolin alone also triggered slow waves and bursts of spikes, and increased insulin release. Both effects were larger in the presence of arginine, but not in the presence of alanine. Forskolin considerably increased the electrical and secretory effects of leucine. 5. A higher concentration of glucose (10 mM) induced slow waves with bursts of spikes in all cells and stimulated insulin release. Alanine, arginine and leucine increased 86Rb+ efflux, electrical activity and insulin release. However, the changes produced by the three amino acids displayed different time course, amplitude and characteristics. Forskolin potentiated insulin release and electrical activity induced by glucose alone. These effects were not augmented by alanine, but markedly amplified by arginine or leucine. 6. Several conclusions can be drawn from this study. The three types of amino acids depolarize the beta cell membrane by different mechanisms and produce distinct patterns of electrical activity. Slow waves with bursts of spikes occur only if a decrease in K+ permeability contributes to the depolarization.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3040965 TI - Control of cell volume and ion transport by beta-adrenergic catecholamines in erythrocytes of rainbow trout, Salmo gairdneri. AB - 1. Trout red cells suspended in an isotonic medium containing beta-adrenergic catecholamines or adenosine 3',5'-phosphate (cyclic AMP) enlarge rapidly to reach a new steady-state volume which is maintained as long as hormone is present. The volume response is not changed by inhibition of the Na+-K+ pump with ouabain. The new steady-state volume was shown to result from a dynamic equilibrium involving the simultaneous functioning of two regulatory processes induced by hormone: a volume increase response that causes cells to enlarge by gaining Na+ and a volume decrease response that causes cells to shrink by losing K+. 2. As previously described, the volume increase response due to NaCl entry, is mediated by the activation by cyclic AMP of a Na+-H+ antiport operating in parallel to Cl(-)-OH- exchanges. In addition, it is shown in this paper that the Na+ uptake is a discontinuous, oscillatory process and that NaCl entry continues for several hours, i.e. as long as hormone is present. 3. The volume decrease response involves a passive, Cl(-)-dependent K+ loss. Na+ cannot use this pathway. The response is blocked by replacement of Cl- by NO3-, by loop diuretics (furosemide, bumetanide) but also by inhibitors of the anion exchanger (4,4' diisothiocyanostilbene-2,2'-disulphonic acid (DIDS), niflumic acid). The activation of this ouabain-insensitive, Cl(-)-dependent K+ transport system is not directly triggered by cyclic AMP. It involves an all-or-none type of switching phenomenon which occurs when the cells swell to a certain volume. Thus it is a regulatory response to the increase in cell volume induced by stimulation of the Na+-H+ exchange by cyclic AMP. Inactivation is also volume dependent: when the cell size approaches the initial size the pathway shuts off. Thus the controlling mechanism of the K+ pathway acts like a reversible on-off switch that operates around a given volume. Ca2+ was not found to be involved in this control. Cyclic AMP is not necessary to keep the activated K+ pathway open but it could be one of the factors involved in the activating process. 4. There are several lines of evidence indicating that in trout red cells the volume decrease and the volume increase responses may not be brought about by the same transport mechanism operating in different modes. The movements of Na+, K+ and Cl- account for the water movements during volume increase and decrease. Thus movements of other solutes such as amino acids need not be considered. PMID- 3040967 TI - Chronic block of the cervical trunk increases synaptic efficacy in the superior and stellate ganglia of the guinea-pig. AB - 1. The effects of chronic conduction block with tetrodotoxin (TTX) of the cervical sympathetic trunk on synapses in the superior and stellate ganglia were examined in vitro with intracellular recording techniques. 2. The mean maximum amplitude of excitatory post-synaptic potentials (e.p.s.p.s) evoked in superior cervical ganglion neurones by stimulation of the cervical sympathetic trunk was increased significantly after 2 or 4 days of block. The electrical properties of the ganglion cells were not appreciably changed by the period of inactivity. 3. Chronic conduction block for 4 days also resulted in a significant increase in the amplitude of the e.p.s.p.s. evoked in stellate ganglion cells by active collaterals of the blocked fibres. 4. The number of steps in the synaptic response elicited in individual stellate ganglion neurones by graded stimulation of the ansa subclavia and the subclavian trunk (which join to form the cervical sympathetic trunk) was increased after 4 days of conduction block. 5. These results show that elimination of impulse activity in the cervical sympathetic trunk increased synaptic efficacy at the disused synapses in the superior cervical ganglion and at the active synapses in the stellate ganglion. In the latter case, collaterals of the blocked fibres apparently formed new synaptic connexions on stellate ganglion cells. PMID- 3040968 TI - Multiple receptor sites for a molluscan peptide (FMRFamide) and related peptides of Helix. AB - 1. The membrane actions of some tetrapeptide amides and heptapeptide amides chemically related to the molluscan neuropeptides Phe-Met-Arg-Phe-NH2 (FMRFamide) and p-Glu-Asp-Pro-Phe-Leu-Arg-Phe-NH2 (pQDPFLRFamide) were tested and compared on identified neurones of Helix aspersa. 2. The C-terminal sequence -Phe-NH2 was an important requirement for each of the four different actions studied: slow increase in K conductance (gK), fast increase in gK, increase in Na conductance (gNa) and decrease in gK. 3. The response of some neurones involved a combination of such actions. 4. The tetrapeptide amides FMRFamide, Phe-Leu-Arg-Phe-NH2 (FLRFamide) and Phe-Ile-Arg-Phe-NH2 (FIRFamide) were more potent than the heptapeptide amides at producing the slow increase in gK and also produced the increase in gNa, not seen at all with the heptapeptide amides. 5. The heptapeptide amides induced the fast increase in gK which was not observed with the tetrapeptide amides. 6. Evidence is presented that each of the tetrapeptide amides tested acts on the same receptor type which mediates the increase in gNa and on which the heptapeptides were inactive. 7. The results are interpreted in terms of multiple receptors each of which however appears to require the C terminal sequence Phe-Met(or Leu)-Arg-Phe-NH2, as has been shown for some molluscan muscle preparations which react to these peptide amides (Price & Greenberg, 1980). PMID- 3040969 TI - Preventive durapatite ridge augmentation for esthetic fixed prosthodontics. AB - A method of preventive ridge augmentation using durapatite granules to prevent residual ridge collapse in the region of fixed partial denture pontics has been described. PMID- 3040970 TI - The effect of surface roughness of crown preparations on retention of cemented castings. PMID- 3040966 TI - The effect of splanchnic nerve stimulation on adrenocortical activity in conscious calves. AB - 1. Right adrenal and various cardiovascular responses to stimulation of the peripheral end of the right splanchnic nerve have been investigated in the presence and absence of exogenous adrenocorticotrophin, ACTH1-24, (5 ng min-1 kg 1). The adrenal-clamp technique was employed in conscious calves in which the pituitary stalk had been cauterized 3-4 days previously. 2. The I.V. infusion of ACTH1-24 increased mean plasma ACTH concentration by about 1200 pg/ml and mean right adrenal cortisol output by about 500 ng min-1 kg-1. Stimulation of the peripheral end of the right splanchnic nerve at 4 Hz for 10 min produced a further rise in cortisol output, amounting to about 400 ng min-1 kg-1 (P less than 0.01). These changes in output were reflected accurately by changes in peripheral plasma cortisol concentration. 3. Closely similar amounts of adrenaline were released in response to splanchnic nerve stimulation in the presence and absence of exogenous ACTH. In the presence of ACTH the average mean output of noradrenaline (58 +/- 2 ng min-1 kg-1) was significantly less than that of adrenaline (102 +/- 4 ng min-1 kg-1; P less than 0.001), whereas the corresponding values were not significantly different in the absence of ACTH. 4. These results also confirm the fact that the fall in adrenal vascular resistance which occurs during splanchnic nerve stimulation is substantially reduced by ACTH, as is the rise in met5-enkephalin output. 5. It is concluded that the splanchnic innervation is capable of enhancing the secretion of adrenal glucocorticoids in response to ACTH under physiological conditions in the conscious calf. PMID- 3040971 TI - Hydroxyapatite-coated subperiosteal dental implants: design rationale and clinical experience. PMID- 3040972 TI - Nicotinic acetylcholine receptors on a cholinergic nerve terminal in the cockroach, Periplaneta americana. AB - Intracellular microelectrode recording and ionophoretic application of carbamylcholine (CCh) were used to compare the cholinergic sensitivity of postsynaptic dendrites of an identified neurone with that of an identified presynaptic cholinergic axon. The axon of the lateral filiform hair sensory neurone (LFHSN) in the first-instar cockroach Periplaneta americana was found to be as sensitive to CCh as the dendritic regions of giant interneurone 3 (GI 3). The CCh response of both neurones was unaffected by replacing Ca2+ with Mg2+, confirming that the ACh receptors are present on the neurones under test. The CCh response of both neurones was mimicked by ionophoretic application of nicotine. The responses were blocked by 10(-5) M mecamylamine and 10(-6) M d-tubocurarine and were not affected by muscarinic antagonists, suggesting that the ACh receptors present on GI 3 and LFHSN are predominantly nicotinic. The muscarinic agonist oxotremorine and the antagonists atropine and quinuclidinyl benzilate had no modulatory effect on LFHSN-GI 3 synaptic transmission. The latency of the LFHSN response to CCh was consistent with the hypothesis that ACh receptors are situated on the main axon/terminal within the neuropil of the ganglion. It has previously been shown that this region of the axon does not form output synapses (Blagburn et al. 1985a). This indirect evidence indicates that presynaptic or extrasynaptic ACh receptors are present in the membrane of a cholinergic axon. LFHSN was depolarized by synaptically-released ACh after normal or evoked spike bursts, suggesting that the nicotinic ACh receptors act as autoreceptors. However, it was not possible to obtain direct evidence to support the hypothesis that these receptors modulate ACh release. PMID- 3040973 TI - Biochemical and immunochemical analysis of avian beta 1 and mammalian beta 2 adrenergic receptors. AB - We have studied the molecular properties of avian beta 1-adrenergic receptor and human beta 2-adrenergic receptor. The turkey erythrocytes beta 1-receptor has been solubilized in active form by digitonin and has been purified to homogeneity by affinity chromatography followed by electroelution from polyacrylamide gel. The photoactivable ligand, iodocyanopindololdiazirine, labels specifically a major 45 kDa and minor 55 kDa polypeptide in turkey erythrocytes, whereas in A431, it labels two polypeptides of molecular weights 65 kDa and 55 kDa. Both types of receptors are N- and possibly O-glycosylated but the turkey beta 1 receptor has only complex carbohydrates whereas the human beta 2 receptor has in addition oligo mannosidic polysaccharidic moiety. Polyclonal and monoclonal antibodies were raised against the beta 1- and beta 2-adrenergic receptors. Polyclonal antibodies were found to mimic beta-adrenergic agonists by stimulating adenylate cyclase upon binding to the receptors. The monoclonal antibodies precipitated both intact and affinity labeled receptors which they also revealed on immunoblots. PMID- 3040974 TI - Purification, biosynthesis and regulation of membrane bound receptors. September 1986, Cap d'Agde, France. Proceedings. PMID- 3040975 TI - Subunit structure and purification of opioid receptors. AB - Considerable evidence indicates the existence of multiple types of opioid receptors. The three major types have been named mu, delta and kappa. The earlier evidence was based on pharmacological as well as membrane binding experiments. This paper will emphasize more recent studies using solubilized opioid binding sites. Several laboratories, including our own, have succeeded in separating kappa receptors from other types. A similar separation of mu from delta receptors has not yet been achieved. By crosslinking experiments with 125I- human beta endorphin we have been able to provide strong evidence for differences in molecular size between the major binding components of mu (65K) and delta (53K) receptors. It is not yet established whether the difference resides in the protein or carbohydrate portion of these glycoproteins. These results suggest that the three major types of opioid receptors represent distinct molecular entities. An active opioid binding protein solubilized from bovine striatal membranes has been purified to apparent homogeneity. The major purification steps involve affinity chromatography and lectin chromatography on immobilized wheat germ agglutinin. The purified material gave a single band of molecular weight 65K Da on SDS-PAGE. Its specific activity for opioid binding was ca. 13,000 pmol/mg protein and its properties are those of a component of the mu receptor. PMID- 3040976 TI - Functional properties and molecular structure of central and peripheral neurotensin receptors. AB - Membranes prepared from mammalian brain or intestine contain two types of specific binding sites for neurotensin that differ by their affinity and by their sensitivity to sodium ions, GTP, and the antihistamine drug levocabastine. Only the high affinity sites are present in cell cultures and in soluble extracts of CHAPS-treated membranes. These sites represent functional neurotensin receptors coupled to GTP-binding proteins that regulate intracellular levels of cAMP, cGMP and inositol phosphates in neuroblastoma N1E115 cells. The molecular weight of neurotensin receptors in cells and membrane preparations of various origin is about 110,000. PMID- 3040977 TI - The role of beta, gamma-subunits of guanine nucleotide binding proteins in control of a reconstituted signal transmission chain containing purified components of the adenylate cyclase system. AB - The properties of a reconstituted signal transmission chain using purified beta 1 adrenoceptor (R), G-protein subunits (G) and adenylate cyclase (C) in lipid vesicles are described. This assay system was used to test beta, gamma-subunits of different origin with respect to their effects on R X G and R X G X C coupling and on the functional properties of GS alpha and Gi alpha. The findings reported here point to large differences in the efficacy of beta, gamma-subunits from different sources assessed by deactivation of [ALF4]-activated rabbit liver GS and pertussis toxin-catalyzed ADP-ribosylation of bovine neutrophil G alpha. This is explained by differences in the interaction domains of the interacting subunits. Furthermore, the sensitivity of R X G and R X G X C coupling to inhibition by beta, gamma-subunits was greater than the effects of beta, gamma subunits on hormonally activated GTPase activity of GS. One of the consequences of differential inhibition of R X G X C coupling is an amplified response of the signal transmission chain to hormonal activation. This is in agreement with observations reported by Cerione et al. PMID- 3040978 TI - Deactivation of photoactivated rhodopsin by rhodopsin-kinase and arrestin. AB - Photoactivated rhodopsin (R) catalyses, by repetitively interacting with many copies of a guanosine nucleotide binding protein (transducin), the amplified binding of GTP to transducin molecules which then activate cyclic GMP phosphodiesterase. Electrophysiologists recently have shown that cyclic GMP keeps ion channels in the plasma membrane of the rod outer segment open in darkness, and that light-induced hydrolysis of cyclic GMP leads to closure of the channels and therefore to hyperpolarization of the rod cell. Photoactivated rhodopsin interacts not only with transducin, but with two more proteins: a protein kinase that specifically phosphorylates R (in contrast to dark-adapted rhodopsin) at multiple sites; and an abundant soluble protein of 48 KDal (called 48 K-protein, S-antigen, or arrestin) that specifically binds to phosphorylated R. Phosphorylation partially suppresses the ability of R to catalyze transducin mediated phosphodiesterase activation even in the absence of arrestin. Binding of arrestin to the phosphorylated R potentiates this inhibitory effect, most probably because arrestin competes with transducin for binding on the phosphorylated R. Phosphorylation, in conjunction with arrestin binding, therefore appears to be a mechanism that terminates the active state of the receptor, R. PMID- 3040979 TI - Metabolism of adrenergic receptors and adenylate cyclase. AB - The alpha 1 and beta-adrenergic receptor metabolism was studied at cell confluency in BC3H1 and C6 glioma cells. After their irreversible blockade with phenoxybenzamine and a bromoacetyl derivative of pindolol (Br-AAM-pindolol) respectively the receptor reappearance allows to determine a half life of 23 hours for the alpha 1-adrenergic receptor in BC3H1 and a quasi absence of beta adrenergic receptor metabolism in C6 glioma cells at confluency. In contrast, beta-adrenergic receptor is rapidly synthesized during cell division. This metabolic stability of beta-adrenergic receptor at confluency was also observed in BC3H1 cells using the heavy isotope labeling of the beta-adrenergic receptor (half life of 8 days). This stability was also confirmed by the observation that at confluency in C6 glioma cells, beta adrenergic receptors reappeared at the cell surface after a complete down-regulation. In parallel with the study of the half life of adrenergic receptors, we determined in BC3H1 the half life of the forskolin stimulated catalytic unit of the adenylate cyclase using heavy isotope labeling method. In heavy amino-acid medium the apparent sedimentation coefficients of the adenylate cyclase increased from 7.4 +/- 0.04S (n = 36) to 8.4 +/- 0.03S (n = 13). This increase was due to the synthesis of new heavy molecule since it was blocked by cycloheximide. The analysis of the kinetic of synthesis of heavy molecules allowed to calculate a half life of 36 hours. The comparison between the half life of several regulatory membrane proteins in BC3H1 indicate that each of them has a specific metabolism. PMID- 3040980 TI - Regulation of the protein kinase activity of the human insulin receptor. AB - The insulin receptor is a hormone-dependent protein tyrosine kinase that belongs to the family of tyrosine kinases associated with growth factor receptors and oncogene products. The activity of the insulin receptor kinase is regulated by the phosphorylation state of specific domains of the protein. Phosphorylation of the receptor on tyrosine residues activates its kinase activity whereas phosphorylation on serine and/or threonine residues inhibits it. In this review, we discuss the evidence that supports a role of the kinase activity of the receptor in the molecular mechanism of insulin action. PMID- 3040981 TI - Progress towards the understanding of the GABAA receptor structure. AB - The GABA receptor of mammalian brain is a ligand-gated channel protein with allosteric binding sites for the benzodiazepines and barbiturate drugs. The receptor is an acidic oligomeric membrane glycoprotein and it has been purified to homogeneity from bovine cerebral cortex, bovine cerebellum and rat cerebral cortex by benzodiazepine affinity chromatography. In each case, extraction and purification with the zwitterionic detergent CHAPS and exogenous phospholipid has demonstrated the coexistence of GABA, benzodiazepine and cage convulsant ligand binding sites on a single protein complex; in addition the allosteric interactions between these sites are preserved in the isolated protein. The receptor has a heterologous structure that is conserved at the subunit level between the aforementioned mammalian species and brain regions. SDS-PAGE has shown that the receptor consists of two subunits, alpha (Mr 53000) and beta (Mr 57000) present in equal stoichiometry. A model consistent with the determination of the molecular weight of the native protein, i.e., Mr 230,000, is that of a tetramer alpha 2 beta 2. [3H]Flunitrazepam and [3H]muscimol have been employed as photoaffinity labels to map the benzodiazepine and GABA binding polypeptides respectively. Polyclonal and monoclonal antibodies have been raised to the native bovine GABAA receptor and these have been employed for the further characterisation of the receptor protein. PMID- 3040982 TI - Adaptive responsiveness of some central receptors to the denervation of heterologous afferent fibers: functional significance and possible behavioral consequences. AB - As shown in the rat brain, the development of the classical "denervation supersensitivity" of receptors does not always occur despite the complete destruction of the corresponding afferent fibers. This is due to the concomitant destruction of heterologous fibers which directly or indirectly contribute to the regulation of the denervated receptors. In this review, on the basis of data obtained in rats with lesions of ascending DA neurons, three examples of hetero regulation of receptors will be provided. In the prefrontal cortex, the concomitant destruction of afferent noradrenergic fibers (NA) prevents the development of the denervation supersensitivity of DA receptors of the D1 type. In the nucleus accumbens, the development of the denervation supersensitivity of D1 receptors seems to be dependent on the state of activity of the cortico nucleus accumbens glutamatergic projection. Finally, in cortico-limbic structures, the prolonged interruption of DA transmission leads to an increased density of 125I-neurotensin binding sites. These hetero-regulations reflect the existence of possible functional relationships between the two types of neuronal populations under investigation. As an example, it will be shown that some dysfunctions seen following the degeneration of the mesocortico-prefrontal and mesolimbic DA neurons disappear following the combined destruction of ascending NA neurons. PMID- 3040983 TI - Association of [3H]-imipramine and [3H]-paroxetine binding with the 5HT transporter in brain and platelets: relevance to studies in depression. AB - [3H]-Imipramine and [3H]-paroxetine label with high affinity a site which is associated with the serotonergic transporter in brain and platelets. The pharmacological profile of inhibition by drugs of [3H]-imipramine and [3H] paroxetine binding is highly correlated with the potency of the drugs to inhibit the uptake of 5HT. Denervation of serotonergic neurons by electrolytic lesions or with 5,7-dihydroxytryptamine produces marked decreases in the density of [3H] imipramine as well as [3H]-paroxetine binding. Dissociation kinetic experiments support the view that the substrate recognition site for 5HT is different from the modulatory site which is labelled by [3H]-imipramine or [3H]-paroxetine. The existence of an endogenous ligand acting on the [3H]-imipramine recognition site to modulate the 5HT transporter was proposed by several laboratories. [3H] Imipramine binding in platelets appears to be a biological marker in depression. Studies carried out in several laboratories report a significant decrease in the Bmax of platelet [3H]-imipramine binding without changes in Kd, when severely depressed untreated patients are compared with healthy volunteers matched for age and sex. The Bmax of platelet [3H]-imipramine binding appears to be a state dependent biological marker in depression. It is tempting to speculate that the endocoid of the [3H]-imipramine recognition site may play a role in the pathogenesis of depression. PMID- 3040984 TI - Partial purification and pharmacology of peripheral-type benzodiazepine receptors. AB - This report describes the results obtained with a new photoaffinity ligand for the "peripheral-type" benzodiazepine binding site (PBS), using a digitonin solubilized preparation from rat heart or adrenals. The specific binding activity of the solubilized adrenal preparation is higher than 50 pmol/mg protein, with binding properties and pharmacological specificity identical to the membrane bound PBS. The apparent molecular weight of the solubilized PBS, determined by gel filtration is 215 KDa. The photoaffinity ligand (PK 14105) is a nitrophenyl derivative of PK 11195, which attaches covalently and specifically to all the PBS when cardiac membranes are irradiated with this compound under ultraviolet light. After photolabelling with [3H]PK 14105 and solubilization in SDS of heart or adrenal membranes, gel electrophoresis indicates the existence of a single protein band whose molecular weight (18 KDa) is unaltered by incubation with sulphydryl-reducing or protein cross-linking agents. This molecule seems to be a low molecular weight, acidic protein. Diethylpyrocarbonate decreases partially (60%) the binding of [3H]PK 11195 without affecting [3H] RO5-4864 binding, which implies a vital histidine residue in the binding domain of [3H]-PK 11195. Treatment with phospholipase A2 or mellitin, a stimulant of endogenous PLA2, led to a selective loss of [3H] RO5-4864 binding with no change in the binding of [3H]PK 11195. Such differences between a benzodiazepine ligand and an isoquinoline ligand suggest that these compounds may induce, on binding, different conformational changes in the PBS, which is compatible with the hypothesis that RO5-4864 and PK 11195 may be an agonist and an antagonist respectively at the PBS. PMID- 3040985 TI - Thyrotrophin releasing hormone--5-hydroxytryptamine interactions in the brain studied using chronic immunization and chemical lesioning techniques. AB - This study investigates the effects of chemically lesioning 5-hydroxytryptamine (5HT) neurones and chronic passive immunization of central thyrotrophin releasing hormone (TRH) on 5HT and TRH mediated behavioural responses. 5HT lesions produced by 5,7-dihydroxytryptamine (5,7-DHT) enhanced the behavioural response produced by the 5HT receptor agonist 5-methoxy-N,N-dimethyltryptamine (5-MEODMT) while decreasing the locomotor hyperactivity observed following administration of the TRH analogue CG 3509 but having no effect on the reversal of pentobarbitone sleep time produced by CG 3509. Chronic intracerebroventricular infusion of the purified TRH antibody markedly increased the length of pentobarbitone-induced sleep-time while enhancing the effects of CG 3509 both on locomotor activity and pentobarbitone-induced sleep. TRH antibody infusion also increased the response produced by 5-MEODMT. The results indicate that chronic passive immunization of central TRH induces changes in TRH receptor responsiveness and that there is a functional interaction between TRH and 5HT neuronal systems. PMID- 3040986 TI - Receptor plasticity in the human brain: some autoradiographic studies. AB - Receptor modifications in human postmortem material were studied by quantitative autoradiography. Alterations of several neurotransmitter receptors in neurodegenerative diseases such as senile dementia and Huntington's chorea, in lesions of specific brain pathways, like the visual pathway or after drug treatments, were examined. In all these situations alterions of the density or localization of receptors were seen using autoradiography. The results suggest that several mechanisms of receptor adaptation operate in the human brain. These mechanisms include: compensatory changes in receptor density as a consequence of cell loss, in some cases preceding the neuropathological changes; differential alterations in receptors depending on their location in a given pathway, for example in the visual pathway or selective homologous or heterologous modification of receptors after drug treatment. PMID- 3040987 TI - Immunochemical studies of the muscarinic acetylcholine receptor. AB - Muscarinic receptors have been purified from calf forebrain plasma cell membranes by affinity chromatography on a dexetimide-agarose gel. SDS-PAGE analysis showed a single 70 kDa band. Monoclonal antibodies have been prepared against these affinity purified 70 kDa protein(s). One antibody, M-35, immunoprecipitated up to 80% of digitonin-solubilized muscarinic receptors. M-35 had agonist-like effects on guinea-pig myometrium: it increased the intracellular cyclic GMP content, decreased prostaglandin-induced cyclic AMP accumulation and caused muscle contractions. The two first effects were inhibited by atropine. M-35 was used to visualize muscarinic receptors at the surface of human fibroblastic cells. In the particular cell line used, the receptors have a low affinity for pirenzepine, were negatively coupled to adenylate cyclase and mediated increase in the phosphatidyl-inositol breakdown. PMID- 3040988 TI - Stimulation of collagenase production in human synovial fibroblast cultures by poly (I). poly (C). AB - Poly (I). poly (C) was found to induce collagenase secretion in both "normal" and "rheumatoid" synovial fibroblast cultures. Induction was time dependent, was maximal at 20-50 micrograms/ml poly (I). poly (C) and was dependent on de novo synthesis. Induction was prevented by hydrocortisone (0.1 microgram/ml) but inhibition of prostaglandin E production by diclofenac (0.2 microgram/ml) or ibuprofen (0.1 microgram/ml) did not affect collagenase production. Collagenase induction was accompanied by stimulation of hyaluronic acid and prostaglandin E production. This in vitro system may represent a model for the study of pathological secretory activities in the inflamed joint. PMID- 3040989 TI - Impotence, carpal tunnel syndrome and peripheral neuropathy as presenting symptoms in progressive systemic sclerosis. PMID- 3040990 TI - Regulation of cartilage remodeling by IL-1: evidence for autocrine synthesis of IL-1 by chondrocytes. AB - Evidence is presented for the synthesis of Interleukin-1 (IL-1) by joint synovial tissue and chondrocytes. Purified preparations of mouse and human recombinant forms of this factor stimulate the synthesis of a secretory protease by cartilage. The IL-1 stimulated chondrocyte protease is capable of converting latent collagenase to its active form. Other proteases such as trypsin and the mercurial aminophenyl mercuric acetate will not activate collagenase in the absence of this protease. Evidence is presented showing that chondrocytes synthesize IL-1 suggesting autocrine control of cartilage matrix turnover. PMID- 3040991 TI - The role of proteases in the pathogenesis of osteoarthritis. AB - It is proposed that the cartilage contains enzymes which are responsible for the degradation of the principle components of the matrix, the proteoglycan and collagen. Measurement of acid, lysosomal bound proteases, or neutral proteases shows increases in proportion to the severity of the disease. Collagenase activity also increases in human osteoarthritic cartilage in the same manner. In an experimental model of osteoarthritis, induced by mechanical factors, enzymatic activity also increased. By inhibiting the enzyme activity with chelators, the arthritic process could be slowed. PMID- 3040992 TI - Chondrocyte-mediated breakdown of cartilage. AB - Chondrocyte-mediated degradation of cartilage was studied using bovine nasal cartilage discs cultured for up to 8 days in the presence or absence of chemically defined agents. Breakdown, assessed quantitatively as proteoglycan released into culture medium and qualitatively by gel filtration of (medium and cartilage) products, was potently stimulated by highly purified interleukin-1 (IL 1), bacterial lipopolysaccharides, and prostaglandin F2 alpha. IL-1 action was abolished by anti-IL-1 antibodies but unaffected by hydrocortisone. LPS stimulated breakdown was reversibly inhibited by glucocorticoid hormones, indomethacin, and cAMP. In addition to their usefulness in probing chondrocyte degradative pathways, several of these agents may be of pathogenetic and clinical significance. PMID- 3040993 TI - Collagen degradation by inflammatory phagocytes. AB - Inflammatory phagocytes are frequently found in the osteoarthritic joint. Current models of the pathogenesis of osteoarthritis (OA) would indicate that proteolytic modification of the cartilage extracellular matrix is important in the development of irreversible joint destruction. Neutrophils and macrophages contain collagenolytic proteinases with different substrate specificity and which are capable of degrading native collagens which are present in articular cartilage. The biochemical nature of these proteinases is discussed. Inflammatory phagocytes may contribute to the process of joint destruction in OA. PMID- 3040994 TI - Regulation of collagenase gene expression in synovial cells. AB - The metalloproteinase collagenase that is synthesized and secreted by synovial cells is responsible for the large amount of connective tissue destruction seen in rheumatoid arthritis. We have used a model system of cultured rabbit synovial fibroblasts to better understand mechanisms controlling both the induction and suppression of collagenase synthesis. Induction of collagenase requires an increase in collagenase mRNA and concomitantly, suppression of collagenase synthesis by retinoids and glucocorticoids is accompanied by a decrease in collagenase mRNA. Another metalloproteinase, activator, which is responsible for the activation of latent procollagenase and which has gelatinolytic ability, is coordinately regulated with collagenase. We conclude that there may exist a family of metalloproteinases that is important in the modulation of connective tissues and that is coordinately regulated at the level of mRNA. PMID- 3040995 TI - Adenomatous polyposis arising after the age of 53. PMID- 3040996 TI - Malignant fibrous histiocytoma causing fatal ileal perforation. PMID- 3040997 TI - Bilateral Achilles tendon rupture simulating peripheral neuropathy: unusual complication of steroid therapy. PMID- 3040998 TI - Synthesis and antiviral activity of 9-[4-hydroxy-3-(hydroxymethyl)but-1 yl]purines. AB - Alkylation of 2-amino-6-chloropurine with 5-(2-bromoethyl)-2,2-dimethyl-1,3 dioxane (5) provided 2-amino-6-chloro-9-[2,(2,2-dimethyl-1,3-dioxan-5 yl)ethyl]purine (6) in high yield. This aminochloropurine 6 was readily converted to the antiviral acyclonucleoside 9-[4-hydroxy-3-(hydroxymethyl)but-1-yl]guanine (1) and to its 6-chloro (10), 6-thio (11), 6-alkoxy (12-17), 6-amino (20), and 6 deoxy (21) purine analogues. The guanine derivative 1 was converted to its xanthine analogue 9. Similarly, alkylation of 6-chloropurine with 5 provided a route to 8, the hypoxanthine analogue of 1. Of these 9-substituted purines, the guanine derivative 1 showed the highest activity against herpes simplex virus types 1 and 2 in cell cultures, and in some tests it was more active than acyclovir, with no evidence of toxicity for the cells. A series of monoesters (30 33) and diesters (24-27, 29) of 1 were prepared, and some of these also showed antiherpes virus activity in cell cultures, the most active ester being the dihexanoate 27. PMID- 3040999 TI - Aldosterone antagonists. 2. Synthesis and biological activities of 11,12 dehydropregnane derivatives. AB - Several steroid derivatives having the delta 11-pregnane skeleton with a 17-gamma spirolactone function were synthesized to evaluate their antialdosterone activity and to elucidate the relation between their binding affinity to mineralocorticoid receptor (MR) and their mineralo- and/or antimineralocorticoid activity. Although many of the synthesized compounds showed strong binding affinity for the MR and aldosterone agonist activity, 3-(17 beta-hydroxy-3-oxoandrosta-1,4,6,11-tetraen 17 alpha-yl)propionic acid gamma-lactone exhibited good aldosterone antagonist activity in an in vivo assay. Its in vivo antiandrogenic activity was also found to be relatively weak. PMID- 3041000 TI - Synthesis and evaluation of melphalan-containing N,N-dialkylenkephalin analogues as irreversible antagonists of the delta opioid receptor. AB - N,N-Dialkylated leucine enkephalin analogues containing melphalan (Mel) in place of Phe4 were synthesized as potentially irreversible antagonists of the delta opioid receptor. These compounds, along with the corresponding Phe4 peptides, were tested for both agonist and antagonist activity in the GPI and MVD smooth muscle preparations. All but two of the eight compounds showed antagonist activity at 1 microM against [D-Ala2,D-Leu5]enkephalin (DADLE) in the MVD when tested under reversible conditions; in all cases the Mel4 peptide had lower activity against DADLE than did the corresponding Phe4 peptide. At higher concentrations (10 microM) the two active Mel4 analogues, (benzyl)2Tyr-Gly-Gly Mel-Leu (2a) and (allyl)2Tyr-Aib-Aib-Mel-Leu (3a), both showed weak irreversible antagonism at the delta receptor. Compound 2a was a selective irreversible delta opioid antagonist while 3a was an irreversible antagonist at both the mu and delta opioid receptors. PMID- 3041001 TI - Erythrocyte lysis by monocytes: investigations on the mechanism and role of the target cell hydrogen peroxide catabolizing pathways. AB - The erythrocyte (RBC) lysis by human monocytes incubated with opsonized zymosan particles (OPZ), was inhibited by catalase, chloride-free medium, azide and hypochlorous acid (HOCl) scavengers (taurine, alanine). These findings suggest the requirement for the HOCl-generating myeloperoxidase-hydrogen peroxide chloride system (MPO-H2O2-Cl- system). The HOCl-dependent lysis was increased by inhibiting RBC catalase with aminotriazole (AT). Conversely, the inhibition of RBC glutathione cycle with 1,3-bis (2-chloroethyl)-1-nitrosourea (BCNU) had no detectable effect. Moreover, the recovery of H2O2 and HOCl from OPZ-triggered monocytes was reduced by the presence of RBCs through a process almost completely preventable by pulsing RBCs with AT but not with BCNU. Thus, it appears that RBC targets protect themselves by consuming, primarily via catalase, significant amounts of monocyte-derived H2O2 with a consequent impairment of the HOCl generation. The results suggest a potential role of target cells in modulating the cytolysin production by monocytes. PMID- 3041002 TI - Fast charge translocations associated with partial reactions of the Na,K-pump: I. Current and voltage transients after photochemical release of ATP. AB - Nonstationary electric currents are described which are generated by the Na,K pump. Flat membrane sheets 0.2-1 micron in diameter containing a high density of oriented Na,K-ATPase molecules are bound to a planar lipid bilayer acting as a capacitive electrode. In the aqueous phase adjacent to the bound membrane sheets, ATP is released within milliseconds from an inactive, photolabile precursor ("caged" ATP) by an intense flash of light. After the ATP-concentration jump, transient current and voltage signals can be recorded in the external circuit corresponding to a translocation of positive charge across the pump protein from the cytoplasmic to the extracellular side. These electrical signals which can be suppressed by inhibitors of the Na,K-ATPase require the presence of Na+ but not of K+ in the aqueous medium. The intrinsic pump current Ip(t) can be evaluated from the recorded current signal, using estimated values of the circuit parameters of the compound membrane system. Ip(t) exhibits a biphasic behavior with a fast rising period, followed by a slower decline towards a small quasi stationary current. The time constant of the rising phase of Ip(t) is found to depend on the rate of photochemical ATP release. Further information on the microscopic origin of the current transient can be obtained by double-flash experiments and by chymotrypsin modification of the protein. These and other experiments indicate that the observed charge-translocation is associated with early events in the normal transport cycle. After activation by ATP, the pump goes through the first steps of the cycle and then enters a long-lived state from which return to the initial state is slow. PMID- 3041004 TI - Difference in capacities for virion-to-virion fusion of young and aged HVJ (Sendai virus): a model of membrane fusion. AB - Young and aged HVJ virions differ structurally and morphologically due to changes that occur during aging in vitro or in ovo. Young virions soon after their budding off are rod-shaped, rigid and relatively uniform in size, whereas virions that have aged in vitro after their formation are round, nonrigid and variable in size. These changes during aging seem to be due to the variation of M protein, a "skeletal" protein that is associated with both the envelope membrane proteins and nucleocapsid strands in the virions. The capacities for virion-to-virion fusion of young and aged virions were compared to clarify the relation between the membrane fusion and membrane-associating skeletal proteins. On treatment with polyethylene glycol (PEG), aged virions readily fused, forming large virion vesicles, but young virions were resistant to fusion. Further, aged virions fused even on incubation at 37 degrees C without the fusogen. Thus the capacity for virion-to-virion fusion evidently increases during aging of virions. This result suggests that skeletal proteins associating with the biological membrane are important for preventing membrane fusion, and that virion-to-virion fusion is a good model system for use in studies on the mechanism of membrane fusion. PMID- 3041003 TI - Fast charge translocations associated with partial reactions of the Na,K-pump: II. Microscopic analysis of transient currents. AB - Nonstationary pump currents which have been observed in K+-free Na+ media after activation of the Na,K-ATPase by an ATP-concentration jump (see the preceding paper) are analyzed on the basis of microscopic reaction models. It is shown that the behavior of the current signal at short times is governed by electrically silent reactions preceding phosphorylation of the protein; accordingly, the main information on charge-translocating processes is contained in the declining phase of the pump current. The experimental results support the Albers-Post reaction scheme of the Na,K-pump, in which the translocation of Na+ precedes translocation of K+. The transient pump current is represented as the sum of contributions of the individual transitions in the reaction cycle. Each term in the sum is the product of a net transition rate times a "dielectric coefficient" describing the amount of charge translocated in a given reaction step. Charge translocation may result from the motion of ion-binding sites in the course of conformational changes, as well as from movement of ions in access channels connecting the binding sites to the aqueous media. A likely interpretation of the observed nonstationary currents consists in the assumption that the principal electrogenic step is the E1-P/P-E2 conformational transition of the protein, followed by a release of Na+ to the extracellular side. This conclusion is supported by kinetic data from the literature, as well as on the finding that chymotrypsin treatment which is known to block the E1-P/P-E2 transition abolishes the current transient. By numerical simulation of the Albers-Post reaction cycle, the proposed mechanism of charge translocation has been shown to reproduce the experimentally observed time behavior of pump currents. PMID- 3041005 TI - The organization and sequence of the genes for ATP synthase subunits in the cyanobacterium Synechococcus 6301. Support for an endosymbiotic origin of chloroplasts. AB - The nucleotide sequence has been determined of two regions of DNA cloned from the cyanobacterium Synechococcus 6301. The larger, 8890 base-pairs in length, contains a cluster of seven genes for subunits of ATP synthase. The order of the genes is a:c:b':b:delta:alpha:gamma, b' being a duplicated and diverged form of b. As in the Escherichia coli unc operon, the a gene is preceded by a gene for a small hydrophobic and basic protein. The hydrophobic profile of the potential gene product suggests that its secondary structure is similar to the uncI protein. The smaller DNA fragment, 4737 base-pairs in length, is separated from the larger by at least 15 X 10(3) base-pairs of DNA. It contains a cluster of two genes encoding ATP synthase subunits beta and epsilon. Both clusters of ATP synthase genes are preceded by sequences resembling the -10 (Pribnow) box of E. coli promoters and are followed by sequences able to form stable stem-loop structures that might serve to terminate transcription. These features and the small intergenic non-coding sequences suggest that the clusters are operons, for which the names atp1 and atp2 are proposed. The order of genes within the two clusters is very similar to the gene order in the E. coli unc operon. However, it is most closely related to the arrangement of genes for ATP synthetase subunits a:c:b:alpha and beta:epsilon in two clusters in pea chloroplast DNA. This close relationship between chloroplasts and the cyanobacterium is also evident from comparisons of the sequences of ATP synthase subunits; the Synechococcus proteins are much more closely related to chloroplast homologues than to those in other bacteria or in mitochondria. It is further supported by the cyanobacterial b and b' proteins which, in common with their chloroplast counterpart, subunit I, have extra amino-terminal extensions relative to the E. coli b protein. This extension is known to be removed by post-translational processing in the chloroplast, but its function is obscure. It also seems likely that the cyanobacterial and chloroplast ATP synthases have important similarities in subunit composition. For example, the presence of two related genes, b and b', in the cyanobacterium suggests that its ATP synthase is a complex of nine polypeptides, and that it may have single copies of related b and b' proteins rather than two copies of identical b subunits as found in the E. coli enzyme.4+off PMID- 3041006 TI - Initiation of bacteriophage P22 DNA packaging series. Analysis of a mutant that alters the DNA target specificity of the packaging apparatus. AB - Bacteriophage P22 is thought to package its double-stranded DNA chromosome from concatemeric replicating DNA in a "processive" sequential fashion. According to this model, during the initial packaging event in such a series the packaging apparatus recognizes a nucleotide sequence, called pac, on the DNA, and then condenses DNA within the coat protein shell unidirectionally from that point. DNA ends are generated near the pac site before or during the condensation reaction. The opposite end of the mature chromosome is created by a cut made in the DNA after a complete chromosome is condensed within the phage head. Subsequent packaging events on that concatemeric DNA begin at the end generated by the headful cut of the previous event and proceed in the same direction as the previous event. We report here the identification of a consensus nucleotide sequence for the pac site, and present evidence that supports the idea that the gene 3 protein is a central participant in this recognition event. In addition, we tentatively locate the portion of the gene 3 protein that contacts the pac site during the initiation of packaging. PMID- 3041007 TI - Structure of ubiquitin refined at 1.8 A resolution. AB - The crystal structure of human erythrocytic ubiquitin has been refined at 1.8 A resolution using a restrained least-squares procedure. The crystallographic R factor for the final model is 0.176. Bond lengths and bond angles in the molecule have root-mean-square deviations from ideal values of 0.016 A and 1.5 degrees, respectively. A total of 58 water molecules per molecule of ubiquitin are included in the final model. The last four residues in the molecule appear to have partial occupancy or large thermal motion. The overall structure of ubiquitin is extremely compact and tightly hydrogen-bonded; approximately 87% of the polypeptide chain is involved in hydrogen-bonded secondary structure. Prominent secondary structural features include three and one-half turns of alpha helix, a short piece of 3(10)-helix, a mixed beta-sheet that contains five strands, and seven reverse turns. There is a marked hydrophobic core formed between the beta-sheet and alpha-helix. The molecule features a number of unusual secondary structural features, including a parallel G1 beta-bulge, two reverse Asx turns, and a symmetrical hydrogen-bonding region that involves the two helices and two of the reverse turns. PMID- 3041008 TI - Sodium imbalance as a cause of calcium overload in post-hypoxic reoxygenation injury. AB - In hypoxic-reoxygenation injury, Ca2+ overload is preceded by disturbed Na+ balance, with low activity of the Na+ pump during hypoxia and during reoxygenation. Failure to correct Na+ content rapidly upon reoxygenation might lead to Ca2+ overload by Na+-Ca2+ exchange. This possibility was tested in energy replete myocardium by perfusing with low K+ (0.6 mM) medium to inhibit the Na+ pump throughout a two-stage procedure with low Ca2+ (0.15 mM) in the perfusate, so that Na+ loading occurred prior to excess Ca2+ uptake, as is the case in hypoxia, then with normal Ca2+ (1.3 mM) to allow Ca2+ uptake, as occurs in reoxygenation after hypoxia. Twenty minutes of Na+-loading (stage a) produced cell Na+ and tissue K+ levels similar to those after 40 min hypoxia. In stage b, hearts rapidly developed Ca2+ overload (12.6 +/- 0.90 microns/g dry wt), low ATP (4.8 +/- 0.8 microns/g dry wt), and creatine kinase release (peak 3.5 +/- 1.2 U/min/g dry wt). These values were comparable to those occurring with reoxygenation after 40 min hypoxia (Ca2+ 10.1 +/- 1.09 microns/g dry wt, ATP 6.3 +/- 0.8 microns/g dry wt, creatine kinase peak 2.1 +/- 0.5 U/min/g dry wt). Contractile failure at high resting tension occurred in both groups. In contrast, hearts recovered well from a period of Na+ pump inhibition which was only temporary. This suggests that Na+-Ca2+ exchange could account for Ca2+ overload in reoxygenation injury on the basis of Na+ pump depression developing during hypoxia and sustained in reoxygenation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3041009 TI - Heart Na,K-ATPase activity in cardiomyopathic hamsters as estimated from K dependent 3-O-MFPase activity in crude homogenates. AB - The hamster hereditary cardiomyopathy provides a unique model for the study of membrane abnormalities during chronic congestive heart failure. It is associated with intracellular calcium accumulation, mitochondrial calcification and cell necrosis. Previous studies have shown a decrease in Na,K-ATPase activity purified from ventricle sarcolemma. The present study demonstrates a decrease in K dependent 3-O-methylfluorescein phosphatase (3-O-MFPase) activity from 1.93 to 1.30 mumol/g wet wt. or 33% in crude homogenates from the left ventricle of 7 months-old cardiomyopathic hamsters as compared to control animals. This represents an equivalent decrease in Na, K-ATPase activity. The values are several times higher than previously published for membrane fractions of myocardium from the hamster. Concomitantly, there was an increase in intracellular Na-concentration of the myocardium of 42% whereas the K concentration was unchanged. The decrease in Na,K-pump concentration may be of importance for the increase in intracellular sodium and ensuing calcifying necrosis observed in the myocardium of cardiomyopathic hamsters. It is emphasized that quantification of the Na,K-ATPase or Na,K-pump should preferably be performed using crude homogenates. PMID- 3041011 TI - The sequence of the 3.3-kilobase repetitive element from Dipodomys ordii suggests a mechanism for its amplification and interspersion. AB - DNA from the kangaroo rat, Dipodomys ordii, contains a 3.3-kb, highly repeated sequence that is interspersed throughout the genome in small tandem clusters. One 3.3-kb unit has been cloned into pBR322 and the nucleotide sequence determined. The clone used was shown to be representative of the bulk of such sequences found in the genomic DNA. The sequence contains 10 homologous subunits each ca. 260 bp in length. Comparison of these to one another yielded a 258-bp consensus sequence containing a 35-bp terminal inverted repeat. Two unique stretches also occur. One of these contains a region that could serve as a promoter for RNA polymerase III; the other contains a sequence related to the ARS sequences of yeast. It is proposed that an ancestral sequence similar to the consensus sequence was amplified to 10 or more units, and that, subsequently, two other sequences were inserted. The properties of these insertions may have led to the dispersal of the sequence throughout the genome. PMID- 3041010 TI - Ubiquitin genes as a paradigm of concerted evolution of tandem repeats. AB - Ubiquitin is remarkable for its ubiquitous distribution and its extreme protein sequence conservation. Ubiquitin genes comprise direct repeats of the ubiquitin coding unit with no spacers. The nucleotide sequences of several ubiquitin repeats from each of humans, chicken, Xenopus, Drosophila, barley, and yeast have recently been determined. By analysis of these data we show that ubiquitin is evolving more slowly than any other known protein, and that this (together with its gene organization) contributes to an ideal situation for the occurrence of concerted evolution of tandem repeats. By contrast, there is little evidence of between-cluster concerted evolution. We deduce that in ubiquitin genes, concerted evolution involves both unequal crossover and gene conversion, and that the average time since two repeated units within the polyubiquitin locus most recently shared a common ancestor is approximately 38 million years (Myr) in mammals, but perhaps only 11 Myr in Drosophila. The extreme conservatism of ubiquitin evolution also allows the inference that certain synonymous serine codons differing at the first two positions were probably mutated at single steps. PMID- 3041012 TI - Cocaine dependence treatment on an inpatient detoxification unit. AB - A psychoanalytic-developmental treatment model includes individual and group psychotherapy which enables compulsive freebase cocaine smokers to articulate explanations for their drug dependence. Relapse prevention education includes recommendations for aftercare treatment. Psychosocial history and DSM-III diagnostic data (N = 31) reveals histories of dysfunctional family dynamics, high rates of depressive disorders, and personality disorders. A case history illustrates application of our treatment model and problems in currently available post-detox treatment. Residential therapeutic communities (TCs) need to offer variable program lengths and specialized crack residences in therapeutic milieus with trained clinical staff. TC programs should offer inpatient psychotherapy, family therapy, and provide direct entrance into program affiliated outpatient services. Recommendations for outpatient services include adjuncts to established anonymous self-help group networks in order to reduce changes of relapse. PMID- 3041013 TI - Sonography of ovarian fibromas/thecomas. AB - The sonographic findings in 14 patients with ovarian fibromas/thecomas are described. A broad spectrum of sonographic features are presented and include hypoechoic mass with posterior shadowing (two cases); anechoic mass with good through transmission (with septations, two cases; without septations, four cases), echogenic mass with well-defined posterior wall (three cases); calcified mass (two cases); mixed echogenicity mass (one case). The pattern of a hypoechoic adnexal mass with acoustic shadowing should still suggest a fibroma/thecoma, but in most cases the appearance is nonspecific. PMID- 3041014 TI - Successful infection of the common marmoset (Callithrix jacchus) with human varicella-zoster virus. AB - The common marmoset, Callithrix jacchus, can be infected with human varicella zoster virus (VZV), both wild-type strain KMcC and attenuated vaccine strain Oka/Merck. Infection was accomplished with either whole-cell-associated or cell extract VZV by combined oral-nasal-conjunctival application and was characterized by substantial and persistent anti-VZV antibody responses. The infectivity of VZV for marmosets was destroyed by treatment of inocula with heat or UV light. Diluted inocula with as few as 40 PFU/ml were infectious for marmosets. The lungs were demonstrated to be a major site of viral replication; both the presence of viral antigens and signs of pneumonia were demonstrated in lung tissues. Four serial passages of VZV KMcC were carried out in C. jacchus by a process of in vitro isolation and culturing of VZV from infected lung tissue and reapplication of the cultured isolates to fresh animals. The isolated viruses were identified as VZV both serologically and by restriction endonuclease analyses. The C. jacchus infectivity model should prove useful for determining the efficacy of subunit and live recombinant VZV vaccines as well as for the study of zoster. PMID- 3041015 TI - Analysis of pseudorabies virus glycoprotein gIII localization and modification by using novel infectious viral mutants carrying unique EcoRI sites. AB - We have constructed two pseudorabies virus (PRV) mutants, each with a unique EcoRI restriction site in the nonessential gIII envelope glycoprotein gene. Since no natural PRV isolate has been reported to contain EcoRI sites, the isolation and single-step growth curve analysis of these mutants established that PRV can carry such a site with little ill effect in tissue culture. Virus carrying these defined mutations produced novel gIII proteins that enabled us to begin functional assignment of protein localization information within the gIII gene. Specifically, one viral mutant contained an in-frame synthetic EcoRI linker sequence that was flanked on one side by the first one-third of the gIII gene and on the other side by the last one-third of the gene. The resulting protein lacked the middle one-third of the parental species, including five of eight putative N linked glycosylation signals, but was still glycosylated and found in enveloped virions; it was not secreted into the medium. A second viral mutant contained an in-frame synthetic EcoRI linker sequence that additionally specified a nonsense codon at position 158, producing a gIII protein that was glycosylated and secreted into the medium; the fragment was not found in enveloped virions. By endoglycosidase and pulse-chase analyses, we established a precursor-product relationship between the various forms of gIII expressed in the parental and mutant strains, and perhaps determined certain features of the gIII protein that are required for its efficient export within the cell. PMID- 3041016 TI - Effects of position and orientation of the 72-base-pair-repeat transcriptional enhancer on replication from the simian virus 40 core origin. AB - A number of recent studies have reported that in papovaviruses such as simian virus 40 (SV40) and polyomavirus, the replication of the viral DNA in vivo is activated by the viral transcriptional enhancer or promoter sequences. Both viral and cellular transcriptional enhancers are well known for their ability to activate transcription in a position- and orientation-independent manner. In the present study, we investigated the effect of the position and orientation of the SV40 72-base-pair (bp) repeat enhancer on its replication activation function. We constructed plasmids containing one copy each of the SV40 core origin and enhancer placed in either order and orientation and at different distances from each other. We assayed the replication efficiencies of these plasmids in the presence of an internal control plasmid in COS-1 monkey kidney cells producing the SV40 T antigen required for replication. We found that the 72-bp repeat was capable of activating replication equally well in either orientation when placed 8 or 9 bp from the core origin. The activation of replication was totally abolished, and replication efficiencies in most instances were found to be lower than that obtained with the core origin alone, when the 72-bp repeat was separated from the core origin by distances of 99 bp or more. This was in direct contrast to the situation with polyomavirus, in which activation of replication by the homologous enhancer or by the SV40 72-bp repeat enhancer is known to be position independent. We also found that when the SV40 core origin and the 72-bp repeat enhancer were adjacent to each other, efficient activation of replication was obtained only if the end of the core origin containing the 17-bp A + T block was linked with the enhancer. In the other orientation of the core origin, activation of replication was either diminished or abolished. Hypotheses such as alteration of chromatin structure by the enhancer and interaction between trans acting factors binding to the enhancer and the core origin mediating the activation effect are discussed. PMID- 3041017 TI - Mutants of the Rous sarcoma virus envelope glycoprotein that lack the transmembrane anchor and cytoplasmic domains: analysis of intracellular transport and assembly into virions. AB - The envelope glycoprotein complex of Rous sarcoma virus consists of a knoblike, receptor-binding gp85 polypeptide that is linked through disulfide bonds to a membrane-spanning gp37 spike. We used oligonucleotide-directed mutagenesis to assess the role of the hydrophobic transmembrane region and hydrophilic cytoplasmic domain of gp37 in intracellular transport and assembly into virions. Early termination codons were introduced on either side of the hydrophobic transmembrane region, and the mutated env genes were expressed from the late promoter of simian virus 40. This resulted in the synthesis of glycoprotein complexes composed of a normal gp85 and a truncated gp37 molecule that lacked the cytoplasmic domain alone or both the cytoplasmic and transmembrane domains. The biosynthesis and intracellular transport of the truncated proteins were not significantly different from those of the wild-type glycoproteins, suggesting that any protein signals for biosynthesis and intracellular transport of this viral glycoprotein complex must reside in its extracellular domain. The glycoprotein complex lacking the cytoplasmic domain of gp37 is stably expressed on the cell surface in a manner similar to that of the wild type. In contrast, the complex lacking both the transmembrane and cytoplasmic domains is secreted as a soluble molecule into the media. It can be concluded, therefore, that the transmembrane domain alone is essential for anchoring the RSV env complex in the cell membrane and that the cytoplasmic domain is not required for anchor function. Insertion of the mutated genes into an infectious proviral genome allowed us to assess the ability of the truncated gene products to be assembled into virions and to determine whether such virions were infectious. Viral genomes encoding the secreted glycoprotein were noninfectious, whereas those encoding a glycoprotein complex lacking only the cytoplasmic domain of gp37 were infectious. Virions produced from these mutant-infected cells contained normal levels of glycoprotein. The cytoplasmic tail of gp37 is thus not required for the assembly of envelope glycoproteins into virions. It is unlikely, therefore, that this region of gp37 interacts with viral core proteins during the selective incorporation of viral glycoproteins into the viral envelope. PMID- 3041018 TI - Sequences of herpes simplex virus type 1 that inhibit formation of stable TK+ transformants. AB - We have identified two regions of the herpes simplex virus type 1 (HSV-1) genome that inhibit DNA-mediated transformation of thymidine kinase-less L (Ltk-) cells by the cloned HSV-1 tk gene. When plasmids containing the EcoRI fragments EK or JK were mixed at 30 fmol/ml with the tk gene and transfected into Ltk- cells, the frequency of transformation was inhibited 80 to more than 90% relative to the control. Of the remaining 10 EcoRI fragments of the HSV-1 genome, 8 were inactive and 2 were weakly active. A 6.1-kilobase PstI subclone between 0.743 and 0.782 map units was isolated from pEK. This clone, pEK-P3P4, exhibited antitransformation activity toward HSV-1 tk and also the bacterial genes gpt and neo. pEK-P3P4 contains the alpha 27 gene, and restriction endonuclease inactivation and subcloning studies established that alpha 27 alone did not inhibit transformation. However, alpha 27 plus sequences both upstream and downstream of alpha 27 did inhibit transformation. In addition, alpha 0 or alpha 4 could substitute for alpha 27 in effecting antitransformation with these sequences. Therefore, an alpha gene and two additional loci in pEK-P3P4 are required for antitransformation. A second antitransforming locus in the reiterated sequences common to EK and JK and distinct from those in pEK-P3P4 was also identified but not characterized in detail. How antitransformation may be an expression of regulation of viral and host cell gene expression is discussed. PMID- 3041019 TI - Primer-dependent synthesis of covalently linked dimeric RNA molecules by poliovirus replicase. AB - Poliovirus-specific RNA-dependent RNA polymerase (replicase, 3Dpol) was purified from HeLa cells infected with poliovirus. The purified enzyme preparation contained two proteins of apparent molecular weights 63,000 and 35,000. The 63,000-Mr polypeptide was virus-specific RNA-dependent RNA polymerase, and the 35,000-Mr polypeptide was of host origin. Both polypeptides copurified through five column chromatographic steps. The purified enzyme preparation catalyzed synthesis of covalently linked dimeric RNA products from a poliovirion RNA template. This reaction was absolutely dependent on added oligo(U) primer, and the dimeric product appeared to be made of both plus- and minus-strand RNA molecules. Experiments with 5' [32P]oligo(U) primer and all four unlabeled nucleotides suggest that the viral replicase elongates the primer, copying the poliovirion RNA template (plus strand), and the newly synthesized minus strand snaps back on itself to generate a template-primer structure which is elongated by the replicase to form covalently linked dimeric RNA molecules. Kinetic studies showed that a partially purified preparation of poliovirus replicase contains a nuclease which can cleave the covalently linked dimeric RNA molecules, generating template-length RNA products. PMID- 3041020 TI - Adaptor plasmids simplify the insertion of foreign DNA into helper-independent retroviral vectors. AB - We have previously described several helper independent vector constructions (S. Hughes and E. Kosik, Virology 136:89-99, 1984; J. Sorge and S. H. Hughes, J. Mol. Appl. Genet. 1:547-599, 1982; J. Sorge, B. Ricci, and S. Hughes, J. Virol. 48:667 675, 1983), all of which derive from Rous sarcoma virus. In this report we describe three improvements in the earlier constructions. First, the vectors have been restructured as proviruses, which considerably improves the efficiency of virus production following acute transfection. Second, a series of miniplasmids have been developed, which we call adaptors, and these miniplasmids can be used to convert virtually any DNA segment into a ClaI fragment suitable for insertion into the retroviral (or other) vectors. Adaptors have been developed that supply regions of functional significance, including a splice acceptor and an initiator ATG. Finally, the region of env defining subgroup specificity, A in the original vectors, has been substituted by the corresponding regions of subgroup B and D viruses, giving vectors with additional subgroup specificities and increased host ranges. PMID- 3041021 TI - The mouse mammary tumor virus long terminal repeat directs expression in epithelial and lymphoid cells of different tissues in transgenic mice. AB - A series of transgenic mice was developed that contained the simian virus 40 early region genes under the transcriptional control of the mouse mammary tumor virus long terminal repeat, including the promoter and glucocorticoid response elements. These mice all expressed the transgene in the epithelial cells of a number of different organs, such as lungs, kidneys, and prostate, salivary, and mammary glands, and in Leydig and lymphoid cells. Transcription of the chimeric gene was inducible by glucocorticoids, either after transfection into tissue culture cells or in cells cultured from animals carrying the transgene. Many, but not all, tissues which expressed the simian virus 40 sequences, as determined immunologically and by RNA analysis, developed into tumors, although they showed premalignant features. Since the mouse mammary tumor virus long terminal repeat is expressed in a number of different cell types when inherited through the germ line, the lactating mammary gland-specific transcription of endogenous proviruses must require other factors or sequences to achieve this specificity. PMID- 3041023 TI - Point mutation in the S gene of hepatitis B virus for a d/y or w/r subtypic change in two blood donors carrying a surface antigen of compound subtype adyr or adwr. AB - Genomes of hepatitis B virus (HBV) were cloned from the plasma of a blood donor who carried subviral particles of three distinct subtypes in the following proportions: adr, 25%; ayr, 63%; and adyr, 12%. HBV DNA clones were classified into two groups based on a difference at only one nucleotide in the S gene. Two clones had A as nucleotide 365 that formed part of the codon for lysine as amino acid residue 122 and produced a surface antigen of subtype adr in transfected NIH 3T3 cells. The remaining four clones had G determining the codon for arginine and produced a surface antigen of subtype ayr in transfected cells. Similarly, HBV genomes were cloned from the plasma of an individual who carried subviral particles of subtypes adr (71%) and adwr (29%). Two clones had T and A as nucleotides 476 and 479, respectively. The other seven clones had C and G as the respective nucleotides. Based on a comparison with previously reported HBV genomes of various subtypes, the mutation of nucleotide 479, forming part of the codon for lysine or arginine as amino acid residue 160, was deduced to determine the w or r subtype, respectively. When NIH 3T3 cells were transfected separately with the genome of subtype adw or adr, derived from plasma containing a surface antigen of subtype adwr, and then cocultured, they produced subviral particles of either subtype adw or adr. When cells were transfected with the genomes of subtypes adw and adr simultaneously, however, subviral particles were produced that possessed w and r determinants on the selfsame particles. These results attributed the d/y or w/r subtypic change to a point mutation in the S gene and favored coinfection of hepatocytes with an HBV genome and its mutant as the mechanism of compound subtypes. PMID- 3041022 TI - Nucleotide sequence of a complete mouse intracisternal A-particle genome: relationship to known aspects of particle assembly and function. AB - The 7,095-nucleotide sequence of a mouse genomic intracisternal A-particle (IAP) element, MIA14, is reported. MIA14 is known to be colinear with IAP 35S RNA and to contain functional long terminal repeats. Its internal genetic organization was determined by comparisons with a homologous Syrian hamster element and the related retroviruses simian retrovirus 1 (simian type D) and Rous sarcoma virus (avian type C). MIA14 contains a gag-protease open reading frame of 827 codons and a pol region of 867 codons entered by a frame shift of -1. The env region of 1,100 base pairs has multiple stop codons in all reading frames, consistent with the failure thus far to detect IAP-related glycosylated envelope components. RNA transcribed in vitro from a cDNA clone containing a closely homologous gag protease open reading frame was translated in a cell-free system. The main product was a 73-kilodalton polypeptide immunoprecipitable with antiserum against the authentic IAP gag-related structural protein p73. Rather than ending at the gag-protease boundary, p73 appears to contain 7 to 8 kilodaltons of peptide encoded by the protease domain, a peculiarity possibly related to the observed impairment of normal protein processing in IAPs. The N-terminal 217 codons of gag are unique to murine IAPs and may have been contributed by recombination with a cellular gene. The mouse-specific region of gag encodes a hydrophobic signal peptide with an atypical cleavage site. Delayed cleavage of this peptide could result in anchoring of newly synthesized p73 to the endoplasmic reticulum membrane and restriction of particle assembly to this site. PMID- 3041024 TI - Hepatitis A virus cDNA and its RNA transcripts are infectious in cell culture. AB - A full-length cDNA copy of an attenuated, cell culture-adapted hepatitis A virus (HAV HM-175/7 MK-5) genome was constructed in the PstI site of plasmid vector pBR322. Transfection of monkey kidney cells with this plasmid failed to induce the production of hepatitis A virus (HAV). The HAV cDNA was excised from pBR322 and inserted, without the oligo(dG) X oligo(dC) tails, into an RNA transcription vector to yield plasmid pHAV/7. Transfection of monkey kidney cells with pHAV/7 DNA induced HAV infection. Transfection with RNA transcripts produced in vitro from pHAV/7 yielded about 10-fold more HAV than did transfection with pHAV/7 DNA. Marmosets inoculated with transfection-derived virus developed anti-HAV antibodies and had liver enzyme patterns that closely resembled the liver enzyme patterns seen in animals inoculated with virus from a comparable level of cell culture passage. Infectious RNA transcripts from HAV cDNA should be useful for studying the molecular basis of cell culture adaptation and attenuation as well as for studying specific viral functions. PMID- 3041025 TI - Isolation of a herpes simplex virus cDNA encoding the DNA repair enzyme uracil DNA glycosylase. AB - Activity of the DNA repair enzyme uracil-DNA glycosylase has been shown to increase in herpes simplex virus type 2 (HSV-2)-infected cells. When mRNA derived from either HSV-1- or HSV-2-infected HeLa S3 cells was translated in an in vitro translation system, significant uracil-DNA glycosylase activity could be detected in the lysate. This activity was specific for the removal of uracil from DNA. Lysates from in vitro translation of mRNA derived from uninfected HeLa cells did not contain measurable glycosylase activity. A cDNA library was constructed with mRNA derived from HSV-2-infected cells 10 h postinfection. Pooled isolates from this library were used in hybrid-arrest and in vitro translation reactions to isolate a uracil-DNA glycosylase-specific cDNA. In vitro translation of hybrid selected RNA, by using this cDNA, produced glycosylase activity in the lysate. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of radiolabeled products from this translation reaction showed a protein component with a molecular weight of 39,000. This is consistent with the molecular weight determinations of the purified glycosylase enzyme derived from either uninfected or HSV-infected HeLa cells. Northern (RNA blot) analysis of HSV-derived RNA, by using the glycosylase cDNA as a probe, revealed five overlapping transcripts of 3.4, 2.8, 2.4, 1.7, and 1.0 kilobases. Southern analysis indicated that the DNA sequence encoding the HSV-specific uracil-DNA glycosylase was located between 0.065 and 0.08 map units on the prototypic arrangement of the HSV genome. PMID- 3041026 TI - Overlapping sets of viral RNAs reflect the array of polypeptides in the EcoRI J and N fragments (map positions 81.2 to 85.0) of the Autographa californica nuclear polyhedrosis virus genome. AB - In several parts of the Autographa californica nuclear polyhedrosis virus (AcNPV) genome, nested sets of overlapping RNAs with common 3' or 5' termini have been recognized. In the present report, the pattern of viral transcription and the arrangement of viral gene products in the region of 81.2 to 85.0 map units were investigated. In this segment of the AcNPV genome, at least nine size classes of viral RNA were identified which ranged in size from 1.3 kilobases (kb) to 4.6 kb and exhibited common 3' termini. The detailed restriction map and the nucleotide sequence of this part of the AcNPV genome were determined. Computer analyses revealed several open reading frames (ORFs) on the rightward-transcribed strand with potential TATA and CAAT signals preceding many of the potential ORFs and the 5' termini of some of the mapped RNAs. The leftward-transcribed strand was devoid of major ORFs. The presumptive polypeptides encoded by the larger ORFs ranged in size from 11.3 to 55.6 kilodaltons (kDa). The amino acid sequence of the presumptive polypeptide encoded by ORF3, a 33.6-kDa molecule, exhibited an unusual, clustered 16-fold repeat of the dipeptide arginine-serine in a protein that showed an overall preponderance of basic amino acids. The results of in vitro translation experiments with hybrid-selected RNAs homologous to internal subfragments of the 81.2- to 85.0-map-unit region yielded polypeptides of approximately 28, 34 to 36, and 48 to 50 kDa, which were close in size to the lengths of the major ORFs derived from the nucleotide sequence. The localizations of individual size classes of RNAs in the 81.2- to 85.0-map-unit region of the viral genome were determined precisely at the 3' and 5' termini by S1 protection analyses. Within a sequence of eight nucleotides, all RNAs had the same 3' terminus, which lay close to multiple polyadenylation signals. The initiation sites of the nine different RNA size classes were precisely mapped. As the cap sites of the smaller RNAs (less than 1.8 kb) were determined by S1 protection analyses, a multitude of RNA initiation sites became apparent. It was also shown that the different RNA size classes in the 81.2- to 85.0-map-unit region were detectable as early as 2 h and at least until 36 to 48 h after infection. In unselected cytoplasmic RNA, the size classes of viral RNAs specific for the EcoRI J fragment were detectable early as well as late after infection, although at early times the larger RNAs were detectable in smaller amounts.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3041027 TI - Transforming and mitogenic effects of avian leukemia virus E26 on chicken hematopoietic cells and fibroblasts, respectively, correlate with level of expression of the provirus. AB - We have investigated the effect of E26, an avian leukemia retrovirus, on the growth properties of chicken embryo fibroblasts (CEFs). E26-infected CEFs were not transformed, according to several transformation parameters, but exhibited an activated growth in vitro. They started to grow without latency in serum supplemented medium, maintained long-term growth in regular or low-serum medium, and could grow when seeded at low cell density in low-serum medium. We compared the integration and the level of expression of the proviral DNA in E26-infected CEFs and E26-transformed hematopoietic cells. An average of two provirus copies were found in each kind of cells. However, whereas high contents of both viral mRNA and E26-specific protein products were found in transformed hematopoietic cells, we detected only low amounts of viral mRNA and no E26 protein in infected CEFs. These data show that the level of expression of the E26 provirus is lower in CEFs than in hematopoietic cells. They suggest that transformation efficiency of the virus depends on its level of expression. PMID- 3041028 TI - Induction of simian acquired immune deficiency syndrome (SAIDS) with a molecular clone of a type D SAIDS retrovirus. AB - We have isolated a molecular clone of the full-length integrated provirus of simian acquired immune deficiency syndrome retrovirus serotype 1 (SRV-1) from a fatal case of simian acquired immune deficiency syndrome in a juvenile rhesus macaque. An integrated SRV-1 provirus was cloned, sequenced, and found to contain four large open reading frames encoding gag-precursor protein, protease, polymerase, and envelope. The proviral clone was transfected into D17 canine osteosarcoma cells and found to produce infectious virus. A comparison of the sequences of this clone with a noninfectious clone showed 20 differences, resulting in 10 amino acid changes. Also, a cluster of exchanges, short insertions, and deletions in the 5' leader sequences resulted in extension of the tRNA(Lys) primer-binding site from 14 to 19 nucleotides. Virus isolated from transfected cells was shown to be infectious and pathogenic, resulting in disease that followed the same time course and mortality as disease induced by uncloned, in vitro cultivated virus isolated from diseased animals. These results unequivocally show that a type D retrovirus (SRV-1) causes a fatal immunosuppressive syndrome in rhesus monkeys. PMID- 3041029 TI - Transforming potential of a myc-containing variant of feline leukemia virus in vitro in early-passage feline cells. AB - We studied a naturally occurring variant of feline leukemia virus (FeLV) in which the oncogene myc has substituted for a portion of the viral structural genes (myc FeLV). myc-FeLV was rescued by replication in the presence of FeLV as helper, and its biological activity was examined in early-passage feline cells in vitro. Infection of leukocytes from peripheral blood, spleen, or thymus, or of kitten fibroblasts did not immortalize these cells or alter them morphologically. Northern blot (RNA blot) analysis of virion RNA prepared from the supernatant of infected cells demonstrated the 8.2-kilobase genome of FeLV, but did not demonstrate the 5.0-kilobase genome of myc-FeLV. Apparently, the myc-FeLV genome was lost in the absence of the selective pressure of transformation. In contrast, infection of embryonic fibroblasts with myc-FeLV(FeLV) rendered these cells capable of greatly increased, if not infinite, proliferative potential. The cells were morphologically altered compared with controls and were only loosely adherent to the substrate. The cells failed to proliferate in semisolid medium and did not form tumors when inoculated subcutaneously into athymic mice. Blot analyses demonstrated the presence and expression of integrated proviral DNAs of both FeLV and myc-FeLV in these cells. They appear, then, to represent cells partially transformed by infection with myc-FeLV(FeLV). The action of feline v myc in early-passage cells in vitro was compared to that of avian v-myc. PMID- 3041030 TI - Diverse wild mouse origins of xenotropic, mink cell focus-forming, and two types of ecotropic proviral genes. AB - We analyzed wild mouse DNAs for the number and type of proviral genes related to the env sequences of various murine leukemia viruses (MuLVs). Only Mus species closely related to laboratory mice carried these retroviral sequences, and the different subclasses of viral env genes tended to be restricted to specific taxonomic groups. Only Mus musculus molossinus carried proviral genes which cross reacted with the inbred mouse ecotropic MuLV env gene. The ecotropic viral env sequence associated with the Fv-4 resistance gene was found in the Asian mice M. musculus molossinus and Mus musculus castaneus and in California mice from Lake Casitas (LC). Both M. musculus castaneus and LC mice carried many additional Fv-4 env-related proviruses, two of which are common to both mouse populations, which suggests that these mice share a recent common ancestry. Xenotropic and mink cell focus-forming (MCF) virus env sequences were more widely dispersed in wild mice than the ecotropic viral env genes, which suggests that nonecotropic MuLVs were integrated into the Mus germ line at an earlier date. Xenotropic MuLVs represented the major component of MuLV env-reactive genes in Asian and eastern European mice classified as M. musculus molossinus, M. musculus castaneus, and Mus musculus musculus, whereas Mus musculus domesticus from western Europe, the Mediterranean, and North America contained almost exclusively MCF virus env copies. M. musculus musculus mice from central Europe trapped near the M. musculus domesticus/M. musculus musculus hybrid zone carried multiple copies of both types of env genes. LC mice also carried both xenotropic and MCF viral env genes, which is consistent with the above conclusion that they represent natural hybrids of M. musculus domesticus and M. musculus castaneus. PMID- 3041031 TI - Isolation and characterization of temperature-sensitive mutants of adenovirus type 7. AB - Fifty temperature-sensitive mutants, which replicate at 32 degrees C but not at 39.5 degrees C, were isolated after mutagenesis of the vaccine strain of adenovirus type 7 with hydroxylamine (mutation frequency of 9.0%) or nitrous acid (mutation frequency of 3.8%). Intratypic complementation analyses separated 46 of these mutants into seven groups. Intertypic complementation tests with temperature-sensitive mutants of adenovirus type 5 showed that the mutant in complementation group A failed to complement H5ts125 (a DNA-binding protein mutant), that mutants in group B and C did not complement adenovirus type 5 hexon mutants, and that none of the mutants was defective in fiber production. Further phenotypic characterization showed that at the nonpermissive temperature the mutant in group A failed to make immunologically reactive DNA-binding protein, mutants in groups B and C were defective in transport of trimeric hexons to the nucleus, mutants in groups D, E, and F assembled empty capsids, and mutants in group G assembled DNA-containing capsids as well as empty capsids. The mutants of the complementation groups were physically mapped by marker rescue, and the mutations were localized between the following map coordinates: groups B and C between 50.4 and 60.2 map units (m.u.), groups D and E between 29.6 and 36.7 m.u., and group G between 36.7 and 42.0 m.u. or 44.0 and 47.0 m.u. The mutant in group A proved to be a double mutant. PMID- 3041032 TI - A recombinant plasmid from which an infectious adeno-associated virus genome can be excised in vitro and its use to study viral replication. AB - A recombinant plasmid carrying an infectious adeno-associated viral genome was constructed that differs in several key respects from previously described recombinants. First, the vector is pEMBL8(+), which allows isolation of viral plus and minus strands. Second, the inserted viral sequences contain two XbaI cleavage sites that flank the viral coding domain. These inserts do not affect replication of the virus, and they allow nonviral sequences to be easily inserted between the cis-acting terminal repeats of adeno-associated virus. Third, the viral genome is flanked by PvuII cleavage sites that allow the entire, infectious viral chromosome to be excised from plasmid sequences in vitro. Viral DNA was replicated more efficiently within adenovirus-infected 293 cells if it was excised from the vector with PvuII before transfection. Presumably, the increased efficiency reflects bypass of the excision step which must normally precede replication when a recombinant plasmid enters the nucleus. The ability to bypass the excision step was exploited to search for a viral function required specifically for excision of the viral genome from the integrated state. None of the mutants tested identified a gene product required for excision that was not also essential for replication. The ability to produce pure populations of viral plus and minus strands was used to demonstrate that both strands are infectious. PMID- 3041033 TI - CD8-positive T lymphocytes specific for murine cytomegalovirus immediate-early antigens mediate protective immunity. AB - We have shown in a murine model system for acute, lethal cytomegalovirus (CMV) disease in the immunocompromised natural host that control of virus multiplication in tissues, protection from virus-caused tissue destruction, and survival are mediated by virus-specific CD8+ CD4-T lymphocytes. Protection from a lethal course of disease did not result in a rapid establishment of virus latency, but led to a long-lasting, persistent state of infection. The CD8- CD4+ subset of T lymphocytes was not effective by itself in controlling murine CMV (MCMV) multiplication in tissue or essential for the protective function of the CD8+ CD4- effector cells. The antiviral efficacy of the purified CD8+ CD4- subset was not impaired by preincubation with fibroblasts that presented viral structural antigens, but was significantly reduced after depletion of effector cells specific for the nonstructural immediate-early antigens of MCMV, which are specified by the first among a multitude of viral genes expressed during MCMV replication in permissive cells. Thus, MCMV disease provides the first example of a role for nonstructural herpesvirus immediate-early antigens in protective immunity. PMID- 3041034 TI - Epstein-Barr virus gene expression in P3HR1-superinfected Raji cells. AB - The pattern of Epstein-Barr virus (EBV) RNAs expressed in Raji cells superinfected with P3HR1 EBV was examined. RNAs whose expression was of an immediate-early type (resistant to treatment of the cells with anisomycin) were identified. These RNAs, encoding the EBV reading frames BZLF1 and BRLF1, were probably expressed from defective virus within the P3HR1 preparation, and some of them were responsible for the induction of the EBV productive cycle in the Raji cells. The structures of the B95-8 RNAs equivalent to the anisomycin-resistant RNAs were determined. The RNA encoding the BZLF1 reading frame contained two splices which extended and modified the reading frame from that previously described. PMID- 3041035 TI - Homotypic and heterotypic exclusion of vesicular stomatitis virus replication by high levels of recombinant polymerase protein L. AB - The recombinant polymerase protein L of vesicular stomatitis virus (VSV) expressed in COS cells is able to transcribe and replicate the viral genome, resulting in complementation of temperature-sensitive polymerase mutants of VSV at the restrictive temperature (M. Schubert, G. G. Harmison, C. D. Richardson, and E. Meier, Proc. Natl. Acad. Sci. USA 82:7984-7988, 1985). Here we report that the efficiency of complementation is dependent on the level of L protein expression. Unexpectedly, only cells expressing low levels of recombinant L protein efficiently complemented tsL gene mutants, whereas cells with high levels of L protein did not. In fact, in all cells with high levels of L protein expression, which at 40 h posttransfection represented almost the total number of transfected cells, viral replication not only of the temperature-sensitive mutant but also of wild-type VSV was excluded. The inhibition of VSV appeared to occur at an early stage of the infectious cycle, and wild-type virus of the same serotype (Indiana) as the recombinant L protein as well as wild-type virus of a different serotype (New Jersey) was affected. Measles virus, on the other hand, was not arrested in cells with high levels of recombinant L protein, demonstrating that these cells were still capable of supporting a viral infection. The expression of high levels of only the amino-terminal half of the L protein from a recombinant mutant L gene that contains a small out-of-frame deletion in the middle of the L gene did not inhibit a VSV infection. Since the level of amplification for both L- and truncated L-encoding vectors is similar, we conclude that the arrest of VSV was caused by high levels of functional full length L protein itself and not by high levels of vector-encoded L mRNA or other vector products or by side effects of vector amplification. These data strongly support the idea that the highly conserved gene order of nonsegmented negative strand viruses and the sequential and attenuated mode of transcription are important regulatory elements which balance the intracellular concentration of viral proteins. They both assure that the L gene is the last and the least frequently transcribed gene, giving rise to low levels of L protein necessary for efficient replication. PMID- 3041037 TI - Molecular genetic analysis of a vaccinia virus gene with an essential role in DNA replication. AB - We have identified a gene encoded by vaccinia virus which is essential for DNA replication. The gene, located in the HindIII D fragment of the viral genome, is transcribed early after infection into two transcripts of 3.0 and 3.7 kilobases which share a 3' terminus. The lesions of three temperature-sensitive DNA replication mutants with defects in this gene have been localized by marker rescue with progressively smaller DNA fragments. We have determined by hybrid selection that the gene encodes an 82-kilodalton protein. An antibody has been prepared against this polypeptide and used to quantitate expression of the protein after infection with wild-type virus or with a viral mutant whose lesion maps within this gene. The temporal pattern of expression in the mutant is unaffected, but the product encoded by the mutant is significantly more thermolabile than the wild-type protein. PMID- 3041039 TI - Processing determinants required for in vitro cleavage of the poliovirus P1 precursor to capsid proteins. AB - We generated defined alterations in poliovirus protein-processing substrates and assayed the effects of these alterations with an in vitro expression system. A complete cDNA copy of the poliovirus genome was inserted into a bacteriophage T7 transcription vector. Using this expression template, we produced RNA transcripts containing defined regions of the poliovirus capsid precursor polypeptide (P1) and RNA transcripts containing mutations in the P1 and P2 regions. In vitro translation of P1-derived transcripts allowed us to characterize the 3C-mediated cleavage of P1 to capsid proteins. We demonstrated that, for either posttranslational or cotranslational cleavage at any of the Q-G amino acid pairs within P1, almost the entire P1 precursor is required. We also demonstrated that minimal sequences 3' to the 2A coding sequence are required to generate active 2A proteinase in vitro and that two specific four-amino-acid insertions in protein 2C do not alter 2A- or 3C-mediated processing of the poliovirus polyprotein. In addition, we demonstrated that substantial deletion of P1 sequences does not alter 2A-mediated cleavage of the Y-G site at the P1-P2 junction. These results allowed us to compare the P1 sequences required for 2A- versus 3C-mediated processing of the capsid precursor, and we discuss these results in the context of the three-dimensional structure of the capsid proteins. PMID- 3041036 TI - A cytomegalovirus protein with properties of herpes simplex virus ICP8: partial purification of the polypeptide and map position of the gene. AB - We demonstrated the presence of a single-stranded DNA-binding protein in human cytomegalovirus (CMV)-infected cells with properties analogous to those of herpes simplex virus (HSV) ICP8. Using monoclonal antibody specific for the CMV protein, we analyzed its fluorescence pattern and time of synthesis, mapped the gene encoding it by using a lambda gt11 library of CMV DNA fragments, and monitored its purification by phosphocellulose and DNA-Sepharose chromatography. In all characteristics we examined, the CMV protein behaved analogously to HSV ICP8. Our results are consistent with a functional role of CMV ICP8 in viral replication that is similar to that of HSV ICP8 and with the evolutionary conservation of the gene of interest in two divergent herpesviruses. PMID- 3041038 TI - Dissection of immediate-early gene promoters from herpes simplex virus: sequences that respond to the virus transcriptional activators. AB - The immediate-early promoters of herpes simplex virus give rise to the first series of transcripts after infection. These promoters are composed of compound sequence elements that govern basal level and regulated transcription. The response of three core (truncated) promoters from the herpes simplex virus type 1 IE-4, IE-0, and IE-27 genes to a battery of virus-encoded trans-acting proteins was examined in a short-term transient expression assay system. The results of this study reveal (i) a role for a sequence, 5'---GGGGG---3', flanked by 3 to 5 base pairs of symmetry (the G box), which is present in the upstream region of all immediate-early gene promoters, (ii) a requirement for the consensus sequence protected by ICP4 for autoregulation by this immediate-early gene product, and (iii) an alternative, sequence-independent mechanism for the augmentation of alpha gene expression by the virion-associated transcriptional activator Vmw65, now designated as TIF. PMID- 3041040 TI - Missense mutations in the VP1 gene of simian virus 40 that compensate for defects caused by deletions in the viral agnogene. AB - Simian virus 40 mutants lacking sequences in the late leader region are viable but produce smaller plaques than does wild-type virus. Within three passages at low multiplicities of infection, virus stocks of several such mutants accumulated variants that synthesized an altered form of the major virion protein, VP1, having a slightly faster mobility in sodium dodecyl sulfate-polyacrylamide gels than did the wild-type protein. Because these variants overgrew the original virus stocks, we consider them to be second-site revertants. By construction and characterization of a series of recombinants, the second-site mutations were shown to map to at least two different regions of the VP1 gene. Nucleotide sequence analysis indicated that single-amino-acid changes were responsible for the rapid mobility of VP1. When combined in cis with either a wild-type or mutant leader region, these VP1 mutations sped up by 10 to 20 h the time course of accumulation of infectious progeny but not of viral DNA or VP1. LP1, the protein encoded by the agnogene, was shown previously to be necessary for the efficient transport of the virion proteins to the nucleus or for their efficient assembly with viral minichromosomes. The VP1 missense mutations reported here compensate for the lack of LP1 by facilitating this process. On the basis of these findings and findings reported previously by us and others, we hypothesize that LP1 facilitates the formation of infectious particles by inhibiting the polymerization of VP1 molecules until the time they interact with viral minichromosomes; the VP1 mutations reported here compensate for the loss of LP1 by lessening the potential of VP1 for self-polymerization. PMID- 3041041 TI - Proteolytic processing of foot-and-mouth disease virus polyproteins expressed in a cell-free system from clone-derived transcripts. AB - All picornaviral genes are expressed as a single, large polyprotein, which is proteolytically processed into the system produces functional proteins, including viral protease 3C, which plays a major role in processing the precursor proteins. To study the function of the two putative proteases 3C and leader (L) in processing, we constructed several cDNA plasmids encoding various regions of the FMDV type A12 genome. These plasmids, containing FMDV cDNA segments under the control of the T7 promoter, were transcribed in vitro by using T7 RNA polymerase and then translated in rabbit reticulocyte lysates. The expressed FMDV gene products were identified by immunoprecipitation with specific antisera and analyzed by gel electrophoresis. The results demonstrate the following: (i) the leader protein, L, is processed from the structural protein precursor, P1, in the absence of any P2 or P3 region proteins; (ii) protein 2A remains associated with the structural protein precursor, P1, rather than the precursor, P2; (iii) the processing of the P1-2A/P2 junction is not catalyzed by 3C or L; (iv) the proteolytic processing of polyproteins from the structural P1 region (except VP4/VP2) and the nonstructural P2 and P3 region is catalyzed by 3C. PMID- 3041042 TI - Frequent segregation of more-defective variants from a Rous sarcoma virus packaging mutant, TK15. AB - TK15, a mutant derived from a temperature-sensitive mutant of Rous sarcoma virus (tsNY68), has extremely low infectivity although it has intact viral genes. Previous analyses of the virus and virus-induced transformants showed that the mutant has a defect in packaging of its own genomic RNA, possibly owing to a deletion near the 5' end. Another striking feature of TK15 is that it induces various types of virus-nonproducing (NP) transformants, 15c(-), at high frequency. In this work, the mechanisms of frequent segregation of NP cells were examined by molecular cloning of TK15-derived proviruses from NP cell clones and their sequence analysis. The structure of the major type of provirus, found in about half of the NP cell clones, was colinear with src subgenomic mRNA and was suggested to be due to infection with virions containing subgenomic mRNA in place of genomic RNA. Other types of proviruses present in 15c(-) cells appeared to contain cellular sequences of various lengths replacing various parts of viral sequences. The mechanism for the generation of these proviruses is discussed in relation to the nature of the packaging mutant. PMID- 3041043 TI - Identification and characterization of the human cytomegalovirus immediate-early region 2 gene that stimulates gene expression from an inducible promoter. AB - The human cytomegalovirus (HCMV) XbaI E cloned DNA fragment of approximately 20 kilobases can complement an adenovirus mutant (dl312) defective in the E1a viral gene product (D. J. Spector and M. J. Tevethia, Virology 151:329-338, 1986). This viral DNA fragment contains three immediate-early (IE) genes between 0.709 and 0.751 map units (M. F. Stinski, D. R. Thomsen, R. M. Stenberg, and L. C. Goldstein, J. Virol. 46:1-14, 1983). Two of the IE genes, IE1 and IE2, were isolated and tested for a role in regulating viral gene expression. Since HCMV early and late promoters require additional characterization, the chloramphenicol acetyl transferase (cat) gene, driven by the adenovirus E2 promoter, was used as an indicator of gene expression. cat expression from this heterologous viral promoter was shown to be stimulated by HCMV at early times after infection. The IE1 gene product did not function independently in activating this promoter. The IE2 gene products could independently stimulate the expression of a plasmid of a plasmid when the cat gene was placed downstream of the inducible E2 promoter (E2CAT). Five proteins of different sizes have been predicted to originate from IE2, depending on mRNA splicing. The protein products specified by the IE2 gene were characterized with an antibody to a synthetic peptide according to the open reading frame of exon 2. Three of the five proteins are encoded by exon 2. Three viral proteins of 82, 54, and 28 kilodaltons (kDa) were detected. The exons contained in the region designated as IE2a have open reading frames that could code for two of the smaller proteins of 27 and 30 kDa. This region, when driven by the HCMV enhancer, could independently stimulate gene expression from E2CAT to a high level. A plasmid with the HCMV enhancer upstream of exons, that could code for the HCMV IE2 proteins of 48 and 51 kDa, as well as 27- and 30-kDa proteins, also stimulated E2CAT expression but at a lower level. The activity of this plasmid was augmented by the IE1 gene product, despite the fact that the latter gene product alone was inactive. It is proposed that the HCMV IE region 2 gene products are involved in the regulation of viral or host cell promoters either independently or in combination with other HCMV IE proteins. PMID- 3041044 TI - In vitro recombinants of ground squirrel and woodchuck hepatitis viral DNAs produce infectious virus in squirrels. AB - Hepatitis B viruses of humans, woodchucks, ground squirrels, and ducks are similar biochemically but differ with respect to host range and pathogenicity. To pursue the genetic basis of these properties in the absence of a cell culture system for virus growth, we exploited the demonstrated infectivity of cloned hepatitis B virus DNA in whole animals. We constructed several recombinant molecules in vitro between cloned infectious genomes of woodchuck hepatitis virus (WHV) and ground squirrel hepatitis virus (GSHV) and assayed the recombinants for infectivity after intrahepatic injection in ground squirrels, which support growth of GSHV but not WHV. Two of the recombinants molecules initiated productive infection; in one recombinant genome, 76% of the coding region for the major surface glycoprotein of GSHV and for the overlapping portion of the presumptive gene for DNA polymerase was replaced by WHV DNA; in the other, 29% of the same coding domain was replaced by WHV DNA. These findings demonstrate the feasibility of generating viable recombinants of hepatitis B viruses from different animal species and suggest that the major host range determinants are not encoded within the surface antigen gene of these viruses. PMID- 3041045 TI - Retroviral vector gene expression in F9 embryonal carcinoma cells. AB - When F9 embryonal carcinoma (EC) cells are infected with retroviral vectors, the efficiency of expression of selectable genes is considerably lower than that in mouse fibroblasts infected with the same retroviral vectors. In this study, several retroviral vectors with regulatory sequences placed immediately 5' to a selectable gene were constructed, packaged, and used to infect mouse fibroblasts and F9 EC cells. With selection as an assay, there was a hierarchy of relative expression in F9 cells compared with that in mouse fibroblasts. These internally placed regulatory sequences are the source of the mRNAs detected in F9 EC cells, while both retroviral long-terminal-repeat promoters and internal promoters are the source of steady-state mRNAs in mouse fibroblasts. This effect was observable with both the internally placed herpes simplex virus thymidine kinase promoter and the Moloney murine leukemia virus promoter. PMID- 3041046 TI - Important role of the long terminal repeat of the helper Moloney murine leukemia virus in Abelson virus-induced lymphoma. AB - The helper virus has been shown to play a critical role in the development of lymphoma induced by the defective Abelson murine leukemia virus (A-MuLV). Indeed, A-MuLV pseudotyped with some viruses, such as the Moloney MuLV, has been shown to be highly lymphogenic, whereas A-MuLV pseudotyped with other viruses, such as the BALB/c endogenous N-tropic MuLV, has been shown to be devoid of lymphogenic potential (N. Rosenberg and D. Baltimore, J. Exp. Med. 147:1126-1141, 1978; C. D. Scher, J. Exp. Med. 147: 1044-1053, 1978). To map the viral DNA sequences encoding the determinant of the lymphogenic potential of Moloney MuLV when complexed with A-MuLV, we constructed chimeric helper viral DNA genomes in vitro between parental cloned infectious viral DNA genomes from Moloney MuLV and from BALB/c endogenous N-tropic MuLV. Chimeric helper MuLVs, recovered after transfection of NIH 3T3 cells were used to rescue A-MuLV, and the pseudotypes were inoculated into newborn NIH Swiss, CD-1, and SWR/J mice to test their lymphogenic potential. We found that a 0.44-kilobase-pair PstI-KpnI long terminal repeat-containing fragment from the Moloney MuLV was sufficient to confer some, but not complete, lymphogenic potential to a chimeric virus (p7M2) in NIH Swiss and SWR/J mice, but not in CD-1 mice. The addition of the 3'-end env sequences (comprising the carboxy terminus of gp70 and all p15E) to the U3 long terminal repeat sequences restored the full lymphogenic potential of the Moloney MuLV. Our data indicate that the 3'-end sequences of the helper Moloney MuLV are somehow involved in the development of lymphoma induced by A-MuLV. The same sequences have previously been found to harbor the determinant of leukemogenicity and of disease specificity of Moloney MuLV when inoculated alone. PMID- 3041048 TI - Novobiocin and coumermycin A1 inhibit viral replication and the reactivation of herpes simplex virus type 1 from the trigeminal ganglia of latently infected mice. AB - Herpes simplex virus type 1 was reactivated from the trigeminal ganglia of latently infected mice in a quantitative and time-dependent manner. Novobiocin and coumermycin A1 reversibly inhibited the reactivation of herpes simplex virus type 1. They did not inhibit viral replication in permissive cells (CV-1) but did inhibit replication in cells of neuronal origin (C1300) and acutely infected trigeminal ganglia. PMID- 3041047 TI - v-myb does not prevent the expression of c-myb in avian erythroblasts. AB - The v-myb oncogene of avian myeloblastosis virus transforms myeloid cells exclusively, both in vivo and in vitro. The c-myb proto-oncogene from which v-myb arose is expressed at relatively high levels in immature hematopoietic cells of the lymphoid, erythroid, and myeloid lineages but not in myeloblasts transformed by v-myb. This finding suggested that the nuclear v-myb gene product p48v-myb might act directly to inhibit the normal expression of the c-myb gene. I have therefore used a selectable avian retroviral vector to express p48v-myb in avian erythroblasts which normally express high levels of the c-myb gene product p75c myb. The results demonstrate that p48v-myb and p75c-myb can be coexpressed in the nuclei of cloned cells. Therefore, p48v-myb does not invariably prevent the expression of p75c-myb. PMID- 3041049 TI - Integration of human papillomavirus type 16 DNA sequences: a possible early event in the progression of genital tumors. AB - The keratinocyte line SK-v harbors only integrated human papillomavirus type 16 (HPV 16) DNA sequences, although it originated from vulvar Bowenoid papules predominantly containing multiple copies of free HPV 16 genomes. We have cloned a fragment of cell DNA that contains the integrated HPV 16 DNA sequences and have shown that integration interrupts the HPV 16 genome in open reading frames E2 and L2 and creates a deletion of 813 base pairs. This allows the expression of open reading frames E6 and E7, as actually substantiated by Northern (RNA) blot analysis of SK-v RNAs with subgenomic HPV 16 RNA probes. Using a unique flanking cellular DNA sequence as the probe, we have shown that the integration of HPV 16 sequences had already occurred in the premalignant lesions from which the SK-v cell line was derived. PMID- 3041050 TI - Biomolecular analysis of a defective nontransforming Epstein-Barr virus (EBV) from a patient with chronic active EBV infection. AB - A virus recovered from the saliva of a child with chronic active Epstein-Barr virus (EBV) infection for 8 years was shown to induce EBV early antigen (EBV-EA) in Raji cells and to be expressed into EBV-EA in fresh EBV-negative peripheral blood leukocytes. However, it did not replicate its DNA. Oropharyngeal epithelial cells scraped from recurrent mouth lesions were similarly positive for EBV-EA. DNA extracted from these cells and digested with BamHI contained a 6-kilobase pair fragment homologous to BamHI fragment V and B1 EBV DNA probes. Furthermore, Southern blots of the BamHI and EcoRI digests of the DNA extracted from the cell lines of the patient (transformed with EBV strain B95-8) and of her mother (spontaneous) revealed, in addition to the expected BamHI G, H, H2, and B1 fragments used as probes, additional shorter ones of a presumably endogenous defective virus. PMID- 3041051 TI - Both the rightward and the leftward open reading frames within the BamHI M DNA fragment of Epstein-Barr virus act as trans-activators of gene expression. AB - The BamHI M DNA fragment of Epstein-Barr virus was shown to activate transcription of the cotransfected chloramphenicol acetyltransferase gene under the control of the simian virus 40 early promoter. Both the BamHI-BglII and the HindIII-BamHI subfragments of the BamHI M fragment, corresponding to the rightward reading frame BMRF1 and the leftward reading frame BMLF1, respectively, had the ability to activate transcription from the simian virus 40 promoter. The trans-activating function was well correlated with the expression of nuclear early antigens, which suggests that early antigens encoded by BMRF1 and BMLF1 are responsible for trans-activation and possibly play a role in regulated expression of virus genomes. PMID- 3041052 TI - Expression of the precore region of an avian hepatitis B virus is not required for viral replication. AB - The core-antigen-coding region of all hepadnaviruses is preceded by a short, in phase open reading frame termed precore whose expression can give rise to core antigen-related polypeptides. To explore the functional significance of precore expression in vivo, we introduced a frameshift mutation into this region of the duck hepatitis B virus (DHBV) genome and examined the phenotype of this mutant DNA by intrahepatic inoculation into newborn ducklings. Animals receiving mutant DNA developed DHBV infection, as judged by the presence in hepatocytes of characteristic viral replicative intermediates; molecular cloning and DNA sequencing confirmed that the original mutation was present in the progeny genomes. Infection could be efficiently transmitted to susceptible ducklings by percutaneous inoculation with serum from mutant-infected animals, indicating that infectious progeny virus was generated. These findings indicate that expression of the precore region of DHBV is not essential for genomic replication, core particle morphogenesis, or intrahepatic viral spread. PMID- 3041053 TI - Studies on the origin-specific DNA-binding domain of simian virus 40 large T antigen. AB - The origin-specific DNA-binding domain of simian virus 40 large T antigen was analyzed, and its C-terminal boundary was found to be at or before amino acid 259. This does not include the zinc finger structural motif located at amino acids 302 to 320 (J. M. Berg, Science 232:485-486, 1986). Interestingly, N terminal fragments of 266 and 272 amino acids and larger displayed dramatically reduced origin-binding activity. In addition, the specific DNA-binding properties of truncated proteins purified from both bacterial and mammalian sources were compared. Truncated T antigens from mammalian cells bound specific DNA fragments more efficiently than did their bacterial counterparts. These results implicate posttranslational modification with a role in regulating the DNA-binding activity of large T antigen. PMID- 3041055 TI - Mapping of the gene coding for Epstein-Barr virus-determined nuclear antigen EBNA3 and its transient overexpression in a human cell line by using an adenovirus expression vector. AB - The open reading frame which lies within the Epstein-Barr virus (EBV) T2 cDNA isolated by Bodescot et al. (M. Bodescot, O. Brison, and M. Perricaudet, Nucleic Acids Res. 14:2611-2620, 1986) was inserted into a eucaryotic expression vector containing a strong adenovirus promoter. The T2 cDNA contains viral genomic sequences from the short BLRF3 open reading frame fused to the adjacent BERF1 long open reading frame. After transfection of human cells, the recombinant plasmid directed the expression of a 140-kilodalton protein. The expressed protein had the same molecular weight, subcellular localization, and immunological characteristics as the EBV-determined nuclear antigen EBNA3, which is made in lymphocytes latently infected with EBV. Immunoprecipitation of extracts of transfected cells labeled with [32P]phosphoric acid showed that the EBNA3 protein is phosphorylated. PMID- 3041054 TI - Specific immune serum to the Epstein-Barr virus DNA polymerase. AB - Epstein-Barr virus (EBV) DNA polymerase was released from phorbol ester-treated tamarin (Saguinus oedipus) cells (B95-8) and prepared for use as an antigen by sequential column chromatography with DEAE-Sephadex A-25, DEAE-cellulose, phosphocellulose, and single-stranded DNA cellulose. Proteins from single stranded DNA cellulose with DNA polymerase activity in 100 mM ammonium sulfate were mixed with complete Freund adjuvant and injected intradermally into rats and rabbits. Immune sera that were screened for specific antibody by indirect immunofluorescence procedures reacted with approximately 3% of the cells in EBV producer cultures (B95-8 and P3HR-1) but not with EBV genome-negative cells (BJAB). In functional enzyme assays, immune sera or the immunoglobulin fraction inhibited the activity of purified EBV DNA polymerase 90%. Inhibition of enzyme activity was not affected by absorption of immune sera with insoluble matrices of proteins prepared with tamarin and human cells which lacked the EBV genome. Cellular DNA polymerase alpha was not inhibited by immune sera to the EBV enzyme. PMID- 3041056 TI - Persistent rotavirus infection in mice with severe combined immunodeficiency. AB - Rotaviruses are important pathogens of human infants and the infants of many animal species. The disease produced by these viruses can be described as an acute, self-limiting diarrheal disease, with virus replication localized to the differentiated epithelial enterocytes of the small intestine. Immunologically normal infants shed virus for approximately 5 to 12 days after the onset of infection. Recently, it has been shown that rotavirus can produce a chronic infection in severely immunocompromised children, with virus shedding and intermittent diarrhea lasting from 6 weeks to 2 years (G. A. Losonsky, J. P. Johnson, J. A. Winkelstein, and R. H. Yolken, J. Clin. Invest. 76:2362-2367, 1985; F. T. Saulsbury, J. A. Winkelstein, and R. H. Yolken, J. Pediatr. 97:61-65, 1980). These findings point to an important role for the immune system in recovery from the disease. The study described here examined the outcome of murine rotavirus infection in mice with severe combined B- and T-cell immunodeficiency (SCID) and in immunologically normal seronegative BALB/c mice. Persistent rotavirus infection was established in all mice with SCID which had been inoculated orally as pups. Low levels of virus replication and constant fecal virus shedding characterized the chronic infection. This is the first report of a persistent rotavirus infection in an animal model. PMID- 3041057 TI - Enterovesical fistula secondary to mucinous adenocarcinoma of appendix. AB - We report an unusual case of an enterovesical fistula secondary to adenocarcinoma of the appendix. PMID- 3041058 TI - Leads from the MMWR. Enterovirus surveillance--United States, 1987. PMID- 3041059 TI - Colorado tick fever or ehrlichiosis. PMID- 3041061 TI - The CDC and abortion in HIV-positive women. PMID- 3041060 TI - Leads from the MMWR. Revision of the CDC surveillance case definition for acquired immunodeficiency syndrome. PMID- 3041062 TI - Adenoma of the liver and carcinoid appendix. PMID- 3041063 TI - Is an AIDS vaccine already available? PMID- 3041064 TI - Abnormal response of adrenocorticotropin to corticotropin releasing factor in spontaneously hypertensive rats. AB - The pituitary-adrenocortical response to ovine corticotropin-releasing factor (CRF) was investigated in spontaneously hypertensive rats (SHR) and normotensive Wistar Kyoto rats (WKY) anesthetized with pentobarbital. After intravenous administration of various doses of CRF (0.1, 1.0 and 10 micrograms), the plasma levels of ACTH were significantly increased in both groups. ACTH increments after various doses of CRF were dose-dependent in WKY. ACTH increments after injections of 0.1, 1.0, and 10 micrograms of CRF in SHR were 123.2 +/- 36.5 (SE), 123.0 +/- 52.2 and 60.3 +/- 48.5 pg/ml at 5 min, and 386.3 +/- 73.8, 243.4 +/- 86.6 and 220.0 +/- 31.4 pg/ml at 15 min, respectively. The ACTH increments in SHR at 15 min after administration of 0.1 microgram of CRF were higher (p less than 0.02) than those in WKY. However, ACTH increments in SHR after administration of 10 micrograms of CRF were lower than those in WKY at 5 min (p less than 0.05) and 15 min. These data suggest that the plasma ACTH responses to various doses of CRF represent a dose-unrelated plateau response in SHR. The plasma levels of corticosterone after administration of various doses of CRF did not change significantly and there were no significant differences between the two strains. The results of these experiments suggest that SHR have an abnormal response of the pituitary-adrenocortical axis to CRF, and the abnormal response may be attributable to desensitization of the pituitary to CRF. PMID- 3041065 TI - Inhibitory action on alpha-human atrial natriuretic polypeptide on vascular adrenergic neurotransmission is attenuated in spontaneously hypertensive rats. AB - The purpose of the present study is twofold, firstly to investigate the effects of alpha-human atrial natriuretic polypeptide (alpha-hANP) on norepinephrine overflow from sympathetic nerve endings, and secondly to compare vascular responsiveness in perfused mesenteric preparations in spontaneously hypertensive rats (SHR, Okamoto and Aoki, 7-9 weeks old) and a cohort of Wistar Kyoto rats (WKY). In preliminary studies using normotensive Wistar rats, the pressor responses to electrical nerve stimulation or exogenous norepinephrine application were inhibited by alpha-hANP. Norepinephrine overflow was also suppressed by alpha-hANP, during nerve stimulation. The pressor responses and norepinephrine overflow during nerve stimulation were significantly greater in SHR than in WKY rats. The inhibitory effect of alpha-hANP on these responses was reduced in SHR. These results indicate that alpha-hANP could affect both pre- and post-synaptic sites of the resistance vessels. Further, the reduced inhibition of pressor responses and norepinephrine overflow by alpha-hANP in SHR suggests an insufficient regulation of adrenergic transmission by alpha-hANP in hypertension. PMID- 3041066 TI - [Combination chemotherapy concurrent with small-dose radiation therapy of small cell carcinoma of the lung]. AB - Forty consecutive patients with small cell carcinoma of the lung were treated with chemotherapy, radiotherapy or both. Of 34 patients treated with chemotherapy, 24 were treated with combination chemotherapy, containing cyclophosphamide vincristine methotrexate and procarbazine, concurrent with small dose radiation therapy (500cGy/5 fraction) as a chemosensitizer (COMPrt). The response rate to this regimen was 81% (29% complete) and the 2 year survival rate was 28.6%. These results have been superior to other regimens and the toxicity was not see to be any higher. After completion of COMPrt regimen, 10 patients were treated with intrathoracic radiation therapy (average dose 3000cGy) and 3 received surgical treatment. Radiation therapy improved the 2-year survival rate (42.2%) when compared with those patients who received no radiation therapy (18.2%). Three patients received surgical treatment were considered to be disease free for 23, 17, and 9 months respectively, after induction of chemotherapy. PMID- 3041067 TI - [Two cases of surgically treated triple cancer]. AB - Two cases of surgically treated triple cancer are reported. Case 1: Three tumors, one in the lung, one in the stomach, and one in the colon of a 74-year-old man were diagnosed preoperatively. First a subtotal gastrectomy and transverse colectomy were performed and, second 27 days later, a lobectomy of the lower left lung. Pathologically, each tumor proved to be a primary sion. Case 2: A 70-year old woman, treated surgically for left breast cancer 5 years ago, was found to have anemia. Gastric endoscopy showed a cancer in the antrum. A subtotal gastrectomy was performed. Intraoperatively another tumor in the cecum was found and a right hemicolectomy was also performed. Pathologically each tumor was a primary lesion. PMID- 3041068 TI - [Progressive multifocal leukoencephalopathy in a child with acute lymphocytic leukemia]. PMID- 3041070 TI - [Cyclic AMP]. PMID- 3041069 TI - [Platelet activating factor]. PMID- 3041071 TI - [Acute gastric mucosal lesion (AGML) in patients receiving hepatic artery infusion chemotherapy]. PMID- 3041072 TI - [Prognosis of small hepatocellular carcinoma in relations to the treatment. Study of 100 patients]. PMID- 3041073 TI - [Enhancement of LAK activity with recombinant gamma-IFN in patients with hepatocellular carcinoma]. PMID- 3041074 TI - [A study on the immunologic effects in workers exposed to low levels of toluene diisocyanate (TDI)]. AB - Forty-three workers exposed to low levels of toluene diisocyanate (TDI) during the process of producing polyurethane forms were examined immunologically for IgG, A, M, and E and serum enzyme activities such as serum angiotensin converting enzyme (SACE), serum lysozyme (SLZM) and glycylproline dipeptidyl aminopeptidase (GP-DAP). Air concentration of TDI was annually measured in various places of work during the past five years from 1979 to 1983. The results obtained in the present study were as follows. 1. The air concentration of TDI at all places of work was below the permissible concentration level of 0.02 ppm throughout the study period. 2. Subjective symptoms and abnormal findings on chest X-ray considered directly related to TDI exposure were not observed. 3. No remarkable abnormal findings in blood cell counts and in serum biochemical studies could be seen in any of the workers. 4. The serum IgG levels in workers directly exposed to TDI were significantly higher (p less than 0.05) than those in workers indirectly exposed to TDI and in non-exposed workers. 5. In the study of serum enzymatic activity, SLZM activity in workers exposed directly to TDI was significantly higher (p less than 0.01) than those in workers indirectly exposed to TDI and in non-exposed workers. PMID- 3041075 TI - [Auditory brainstem responses and electroencephalographic findings in patients with occupational vibration disease]. AB - To evaluate the function of the central nervous system of patients with occupational vibration disease, electroencephalograms and auditory brainstem responses (ABR) were recorded in 20 male subjects with occupational vibration disease whose age ranged from 46 to 67 years (mean 57.4 yr). All the subjects had operated chain saws from 10 to 25 yr (mean 15.2 yr) and had frequently complained of many subjective symptoms induced by central nervous system disturbances such as headache, head heaviness, tinnitus, vertigo, and insomnia, which corresponded to stage 3 in the diagnostic criteria of Andreeva-Galanina. Twenty-six healthy men whose age ranged from 40 to 67 yr (mean 53.0 yr) were selected as controls. Electroencephalograms were recorded with a 12-channel electroencephalograph, using unipolar and bipolar leads. Auditory brainstem responses were recorded by signal averaging technique using 100 microseconds alternating clicks. The stimuli were presented at 70 dB above threshold (SL) with a rate of 10 per second. The following results were obtained. 1. By electroencephalography, the incidence of diffuse alpha pattern, slow alpha wave and drowsy pattern was 32%, 32%, and 42%, respectively. 2. Click thresholds in the patients were obviously higher than those in the healthy controls. 3. The occurrence rate of wave II of ABR in the patients (61.8%) was significantly lower than that in the healthy controls (85.0%) (p less than 0.05). 4. The interpeak latencies I-V and III-V of ABR in the patients were significantly longer than those in the healthy controls.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3041076 TI - [Autonomic nervous function in patients with vibration syndrome--an investigation of the R-R interval on the ECG and microvibration on the thenar eminence]. PMID- 3041077 TI - Effects of arildone on the immunogenicity of formalin-inactivated polioviruses. AB - Concentrated and purified Sabin and virulent strains of poliovirus types 1, 2 and 3 were inactivated with formalin at 37 C. By addition of 5.4 microM arildone, an antiviral agent, to the virus suspension, the stability of D antigen increased in both Sabin and virulent strains of all types, especially in virulent type 1 Mahoney strain. The drug had neither any inhibitory nor enhancing effect on the formalin inactivation. When antibody response was compared in guinea pigs, Sabin strains inactivated in the absence of arildone were less immunogenic against homotypic virulent strains than inactivated vaccine prepared from virulent strains. On the other hand, Sabin strains inactivated in the presence of arildone were equally immunogenic. These results indicate that it is possible to prepare from Sabin strains a potent and safe inactivated vaccine having an immunogenicity comparable to that prepared from virulent strains. PMID- 3041078 TI - The handy and simple apparatus for the quantitative determination of hydrogen cyanide in blood. PMID- 3041079 TI - [Evaluation of single photon emission computed tomography using Ga-67 for scintigraphy of hepatocellular carcinoma]. PMID- 3041080 TI - Familial hyper-angiotensin converting enzyme (ACE)-emia: increased production of ACE by monocyte-macrophage. AB - We report here a familial clustering of elevated serum angiotensin converting enzyme (ACE) levels. The patient was a 58-year-old Japanese female. She had been in excellent health until the age of 45, when she noticed a decrease in visual acuity of her left eye. Despite intensive therapy under the diagnosis of occulusion of the central retinal vein, she lost her visual acuity at the age of 45. Thereafter, she has been in excellent health. The only abnormality found in this case has been a markedly elevated level of serum ACE (625 n mol/min/ml; normal range; 22-40 n mol/min/ml of serum). Her blood pressure was within normal limits (140/80 mmHg). There was no evidence for the diagnosis of sarcoidosis, Gaucher's disease, leprosy, hyperthyroidism, diabetic retinopathy, or liver disease. One of her two sisters also showed a marked increase in serum ACE activity (303 n mol/min/ml), and remarkably high levels of serum ACE (276 and 294 n mol/min/ml) were demonstrated in both of two sons of this sister. All the members of this family have been in excellent health. The serum ACE activity was activated by chloride and cobalt ions, and inhibited by EDTA, captopril and rabbit antiserum to purified human plasma ACE. Thus, our study showed a familial clustering of "hyper-ACE-emia", and the disorder appears to have been inherited as an autosomal dominant trait. PMID- 3041081 TI - The clinicopathological significance of changes of ganglioside patterns in the cirrhotic liver: a study of 11 cases. AB - Unique ganglioside components including GM2 and spots No. 1 and 2 designated tentatively were increased in content in some cases of cirrhotic liver as well as in all cases of hepatocellular carcinoma (HCC). Tissues of cirrhotic liver in which these components were increased were examined histologically. These components were increased in cirrhotic liver showing many foci of dysplastic nodules and in cirrhotic nodules at the matrix of multicentric HCC. These findings, thus, indicate that the unique ganglioside components are probably related to the histological findings of the cirrhotic liver. PMID- 3041082 TI - A clinical evaluation of transcatheter arterial embolization on hepatocellular carcinoma. AB - TAE was performed in 49 cases of HCC for clinical assessment of the usefulness. This procedure was aimed to subside intraperitoneal hemorrhage in 8 cases among them and to provide a possible antitumor effect in the remaining 41 cases. The prognosis in the 8 cases of intraperitoneal hemorrhage was 54.9 +/- 41.1 days including one case survived as long as 116 days. The prognosis in the remaining 41 cases was 12.6 +/- 8.3 months in group A, 7.8 +/- 6.1 months in group B and 1.6 +/- 1.3 months in group C according to Child's classification, while it was 2.8 +/- 2.5 months in 1st branch occluded, 9.4 +/- 6.1 months in IInd branch occluded and 19.2 +/- 6.7 months in IIIrd branch occluded group according to the portal vein occlusion due to tumor thrombi suggesting that a more prolonged survival was attained with more favorable degree of Child's classification and less affected portal embolization. The cumulative survival time (by Kaplan Meier's methods) was 6 months in 89% of the cases examined, 1 year in 59%, 2 years in 34% and 3 years or more in 11%, indicating significantly higher survival as compared to our TAI group. Angiographic re-opening of tumor vessel within 3 months was observed in 46.7%. TAE on HCC was useful both for the purpose of antitumor effect and of hemostasis. The degree of Child's classification and severity of portal occlusion at the initial examination may closely relate to the prognosis. Thus, angiography should be repeated within 3 months following the first TAE at least. PMID- 3041083 TI - Biochemical characterization of familial amyloidotic polyneuropathy in various districts of Japan. AB - The purpose of this study is to disclose the molecular basis of type I familial amyloidotic polyneuropathy (FAP) in Japan. Amyloid fibril protein of type I FAP consists of a variant transthyretin (also called prealbumin) with one amino acid substitution of methionine-for-valine at position 30. This variant transthyretin is present in the serum as a precursor protein of amyloid. A radioimmunoassay (RIA) has been established to detect the variant transthyretin. All the 94 patients with FAP who originate from various districts in Japan have the variant transthyretin, but any one of 78 healthy adults of families with FAP do not have it. Half of the symptom-free children of FAP patients have the variant transthyretin even before clinical manifestations appear. The RIA is widely applicable for early diagnosis. The methionine-for-valine substitution is due to a base change from guanine to adenine at the first letter of the valine codon at position 30. Type I FAP in Japan is considered to be a molecular disorder of transthyretin. Since the age of onset ranges from twenties to forties, genetic counseling is recommended to prevent the transmission of this intractable disorder to the next generation. PMID- 3041084 TI - Cases with familiar amyloid neuropathy starting of the upper limbs and having hepatic disorder. AB - Here we are reporting two cases consisting of a male patient and his elder sister from Kagashima in Gifu City and both suffering from polyneuropathy of dissociation type and skin amyloidosis. In the former, the presence of amyloid was demonstrated not only in the skin, but also in the stomach, liver and gums. He was also diagnosed suffering from chronic hepatitis of inactive type. He responded to DMSO and Cepharanthin. In the patients, no urinary Bence Jones protein nor blood M component was detected and the amyloid exhibited resistance to potassium permanganate treatment. The neuropathy of the patients were slightly different from that of the Portuguese type which starts on lower extremities as well as those conventionally have been reported in Japan. PMID- 3041085 TI - Involvement of endogenous noradrenaline release in methylene blue-induced contraction of isolated rabbit aorta. AB - The vasocontractile response to methylene blue (Meb) was investigated in isolated rabbit aorta. Meb (1-100 microM) induced a slowly developing contraction after a long latency in rabbit aortic strip. The maximal contraction was obtained by 50 microM Meb, which corresponded to 1 microM noradrenaline (NA)-induced contraction. Once the maximal contraction was induced by Meb, the strip completely lost the contractile response to a further application of Meb. The usual NA-induced contraction, however, could be observed in such a Meb insensitive aortic strip. Meb-induced contractions were not affected by atropine, diphenhydramine, methysergide, indomethacin, nordihydroguaiaretic acid and removal of endothelial cells from the aortic strip, but they were abolished by prazosin. In aortic strips from rabbits pretreated with reserpine (3.0 mg/kg, i.m.) for a day, Meb failed to induce contraction. Meb evoked the [3H] release from [3H]NA-preloaded aortic strips. In high performance liquid chromatographic analysis, a considerable amount of NA was found in the bathing fluid of the aortic strip in the presence of Meb. In addition, Meb pretreatment inhibited [3H]NA uptake by the aortic strip and abolished the contractile response to an electrical stimulation of adrenergic nerve terminals. Although Meb decreased the basal level of cyclic GMP in the aortic strip, Meb-induced [3H]NA release from the aortic strip was not affected by 8-bromo cyclic GMP. These results suggest that Meb-induced contraction of rabbit aorta is due to the release of endogenous NA from its storage pools of intramural adrenergic nerves through an independent mechanism of its cyclic GMP lowering effect. In addition, incubation of aortic strips with Meb resulted in depleting the storage NA and blocking the nerve function, suggesting that Meb might be useful for a pharmacological tool as an adrenergic neuron blocking agent in vitro. PMID- 3041086 TI - A possible mechanism of protection by polyamines against gastric damage induced by acidified ethanol in rats: polyamine protection may depend on its antiperoxidative properties. AB - The protective mechanism of polyamines against acidified ethanol-induced gastric damage was studied. Their oral administration prevented the formation of gastric mucosal lesions induced by 90% ethanol in 150 mM HCl in a dose-dependent manner, with the order of the protective potency being spermine greater than spermidine greater than putrescine. The acidified ethanol-induced lesions were accompanied by a concomitant increase in gastric mucosal lipid peroxide levels, but spermine in a protective dose could prevent the increment of lipid peroxides. Polyamines, in a concentration-dependent fashion, inhibited the reduction of nitroblue tetrazolium by superoxide anion radicals generated in vitro in the xanthine xanthine oxidase system and the lipid peroxidation in vitro induced by ferrous ion in the porcine gastric mucosal homogenate. The order of the superoxide scavenging potency and the inhibitory potency of iron-induced lipid peroxidation by polyamines corresponded to the order to the protective potency against acidified ethanol-induced gastric lesions. The present results suggest that cytoprotection by polyamines may be responsible for their antiperoxidative activities. PMID- 3041087 TI - Malignant cystosarcoma phyllodes with lymph node metastasis--a case report. AB - A case of malignant cystosarcoma phyllodes with metastasis in the interpectoral lymph node (Rotter's) is presented in this paper. To the author's best knowledge, this is the first case in Japan on this disease, with lymph node metastasis. Although the surgical management of the disease has not yet been standardized, radical or modified radical mastectomy is thought to be the appropriate procedure. PMID- 3041088 TI - [Experimental studies on hepatic circulation: effects of drugs on blood flows in liver tissue and portal vein of the cats]. AB - The effects of some drugs on the hepatic circulation were examined by thermoelectrical and electromagnetic methods under pentobarbital-anesthesia in normal and CCl4-pretreated cats. The following results were obtained. 1. Both adrenergic alpha and beta receptor functions were involved in the regulation of the hepatic circulation of normal cats. 2. Three calcium blockers (nifedipine, nicardipine, diltiazem) had different potencies in increasing the hepatic blood flow of normal cats. 3. The isolated veins including portal vein showed the regional difference in the responsiveness to calcium blockers. 4. In CCl4 pretreated cats, adrenergic alpha receptor function was dominant in the control of hepatic circulation and diltiazem raised portal venous pressure. PMID- 3041089 TI - Circulating immune complex levels in cows with enzootic bovine leukosis. PMID- 3041090 TI - Demonstration of Aujeszky's disease virus antigen in naturally infected cattle by immunoperoxidase method. PMID- 3041091 TI - An ultrastructural observation of esophageal lesion in bovine papular stomatitis. PMID- 3041092 TI - Effect of feline leukemia virus on the development of plaque forming cells in feline peripheral blood lymphocytes. PMID- 3041093 TI - Butyrate-induced cytoarchitectural reorganization of Mallory body-containing rat hepatic tumor cells. AB - Diethylnitrosamine (CAS: 55-18-5)-transformed 72/22 rat hepatic tumor cells undergo marked cytoarchitectural changes during exposure to sodium butyrate in vitro. Butyrate treatment of this cell line resulted in an increased cell size, volume, and protein content and in structural reorganization within both the intermediate filament and microfilament networks resulting in the generation of a more normal appearing hepatocytic phenotype. Induced changes in the microfilament system involved the accumulation of F-actin at the cellular margins in the form of a peripheral band and in the development of an extensive, predominantly centralized network of thickened cytoplasmic filament bundles. Such butyrate induced changes in hepatic tumor cellular morphology and microfilament organization were reflected in a 26-51% increase in the amount of cytoskeletal associated actin in 72/22 cells, as determined by flow cytofluorimetry of permeabilized intact cells or by scanning densitometry of the electrophoretically separated, detergent-resistant cytoskeletal protein fraction, respectively. It is unlikely that this increase in cellular microfilament content was due to a direct effect of butyrate on actin polymerization per se since butyrate (in final concentrations equal to that used in culture) did not alter either actin monomer polymer transitions or the nucleation reaction in a defined in vitro polymerization assay. The available data suggest that butyrate may regulate the synthesis or modulate the actin-binding capacity of microfilament-associating proteins in cultured cells. Butyrate-induced "normalization" of 72/22 cytoarchitecture was previously shown to be reflected in a reduction or loss in the expression of specific growth traits characteristic of the transformed phenotype. The experimental reversal of defined cytoarchitectural abnormalities and transformed growth characteristics of 72/22 cells by butyrate provided an in vitro model to elucidate both particular cytoskeletal events associated with epithelial cell transformation and the mechanism of action of apparent differentiation-inducing agents, such as sodium butyrate, on responsive tumor cells. PMID- 3041094 TI - Enhancement of tumorigenicity with morphological progression in a BALB/c preneoplastic outgrowth line. AB - Four newly established mammary hyperplastic outgrowth lines were analyzed for their tumorigenic, morphological, and ovarian hormone-dependent growth properties in BALB/cMed mice. The mammary outgrowth lines were designated DIM-1, DIM-2, DIM 3, and DIM-4 to indicate their origin from the mammary cell line COMMA-D. DIM-1, DIM-2, and DIM-3 were classical hyperplastic alveolar outgrowth lines that possessed high tumor-producing capabilities and rapidly evolved by transplant generation (TG) 6 into ovarian hormone-independent populations. DIM-3 was also characterized by extensive formation of dilated (cystic) alveoli. This characteristic did not correlate either positively or negatively with the tumor producing capabilities of these lines. DIM-4 was a ductal outgrowth line that exhibited a progression in several biological properties. This line progressed morphologically from a ductal outgrowth to a mixed outgrowth of small ducts (ductules) with scattered alveoli and then in subsequent passages to a uniform alveolar outgrowth. Concurrent with these morphological changes, the tumor producing capabilities of DIM-4 increased from low [40% with the time for 50% of the transplants to produce mammary tumors (TE50) greater than 12 mo] to high (71% with a TE50 of 7.1 mo). The ductal outgrowth line (TG 4) was totally dependent on ovarian hormones for growth; however, later passages (TG 8) of DIM-4 were only partially ovarian hormone dependent for growth and ovarian hormone independent for maintenance of alveolar morphology. In addition, the tumorigenic response to 7,12-dimethylbenz[a]anthracene (CAS: 57-97-6) was low in the ductal stage (TG 4) and high in the alveolar stage (TG 7). The results demonstrate an enhancement of tumorigenicity with morphological and biological progression in a preneoplastic mammary cell population and support the hypothesis that the cells at high risk for tumorigenesis in the BALB/c mammary gland appear to be the alveolar cell type. PMID- 3041095 TI - Origin of the medulloblastoma experimentally induced by human polyomavirus JC. AB - The origin of the medulloblastoma induced by JC virus (JCV) in golden hamsters was investigated by the in situ hybridization method. After inoculation of JCV into newborn hamsters, a few migrating cells in the cerebellar molecular layer as well as several cells in the internal granular layer hybridized with an antisense mRNA probe of JCV T-antigen on the 10th day. The number of cells positive for this probe decreased on the 15th and 20th days. Moreover, an incipient medulloblastoma consisting of many cells positive for T-antigen mRNA was noticed in the cerebellar internal granular layer 30 days after inoculation. About 6 months post inoculation, 95% of the animals had succumbed to medulloblastoma. Therefore, the origin of the medulloblastoma seemed to be the cells in the cerebellar external granular layer that were infected by JCV, that migrated normally through the molecular layer to the internal granular layer, and that began to proliferate to become medulloblastoma. When 5-day-old hamsters were inoculated, a few cells positive for T-antigen mRNA were detected in the cerebellum within a month. In the long-term observation, the ratio of medulloblastoma induction decreased as the age of the animals at inoculation increased. These results support the idea that JCV infects and transforms the cells in the cerebellar external granular layer, because this layer appears only transiently in newborn hamsters. PMID- 3041096 TI - Robert F. Pitts memorial lecture. H+ secretion in renal cortical tubules: kinetic aspects. PMID- 3041097 TI - Reduction of proteinuria by angiotensin converting enzyme inhibition. AB - The effects of the angiotensin converting enzyme (ACE) inhibitor lisinopril on blood pressure, proteinuria and renal hemodynamics were evaluated in 13 patients with renal disease of different origin. A comparison was made with the effects of conventional antihypertensive therapy. Both drug regimens significantly lowered blood pressure, while only after 12 weeks of treatment with lisinopril, blood pressure was significantly lower than during conventional therapy. Lisinopril reduced proteinuria (by 61 +/- 40%), whereas conventional therapy had no significant effect on protein excretion. During the first eight weeks of treatment with lisinopril, there was a comparable degree of blood pressure reduction with both treatment regimens, whereas urinary protein loss was significantly less during ACE inhibition. There was only a nearly-significant positive correlation between the fall in proteinuria during lisinopril and the concomitant decrease in mean arterial pressure. Glomerular filtration rate decreased from 26.3 +/- 11.6 to 20.6 +/- 9.4 ml/min during treatment with lisinopril. This decrease was not correlated with the fall in proteinuria. A significant positive correlation existed between the fall in urinary protein excretion and both the decrease in overall renal vascular resistance, and the fall in filtration fraction. Although blood pressure lowering by itself could contribute to the antiproteinuric effect of lisinopril, our results suggest that this effect of ACE inhibition is also due to efferent (postglomerular) vasodilation. We conclude that the ACE inhibitor lisinopril effectively reduces blood pressure and proteinuria in renal disease. The latter effect is not only the result of a lower blood pressure, but is probably also due to a fall in intraglomerular capillary pressure. PMID- 3041098 TI - Cellular events in vasopressin action. PMID- 3041099 TI - Reversal of toxic and non-toxic effects of digoxin by digoxin-specific Fab fragments in isolated human ventricular myocardium. AB - The time course of the reversal of toxic and nontoxic effects of digoxin by digoxin-specific antibody fragments (Fab) was measured in isolated human ventricular myocardium. A concentration of 2 X 10(-6) mol/l digoxin was used to produce positive inotropy followed by mechanical signs of toxicity. After addition of a 1.5-fold higher molar concentration of digoxin-specific Fab, signs of toxicity disappeared within 30 min and digoxin-induced force of contraction decayed with a monoexponential time course with a half-life of 52 min. This rate of decay was almost identical to that observed for the dissociation of the digoxin-(Na+ + K+)-ATPase complex in human heart cell membranes. It is concluded that digoxin-specific Fab are capable of completely removing digoxin from its binding sites, the maximal rate of removal of digitalis glycosides from the (Na+ + K+)-ATPase is limited by the dissociation rate constant, and there is a close correlation between the degree of binding of digitalis glycosides to the (Na+ + K+)-ATPase and the increase in force of contraction. PMID- 3041101 TI - [Grieg's cephalopolysyndactylia syndrome in one family]. PMID- 3041100 TI - [Urinary steroid profile in Cushing syndrome and in tumors of the adrenal cortex]. AB - The analysis of 24-h excretion profiles of urinary steroids in 18 patients suffering from Cushing's syndrome or adrenocortical tumors revealed typical patterns when compared to 37 healthy control persons, 24 patients with obesity, and 6 patients with hirsutism. The validation of eight criteria--increased excretion of free cortisol, 6 beta-hydroxycortisol, 20 alpha-dihydrocortisol, 11 beta-hydroxyandrosterone, and 3 beta-hydroxy-5-en steroids, decreased ratio of tetrahydrocortisone (THE) to tetrahydrocortisol (THF), and increased ratios of THF to allotetrahydrocortisol (a-THF) and metabolites of androgens (AM) to metabolites of cortisol (CM)--afforded reliable detection of disorders in steroid biosynthesis. The analysis of urinary steroid profiles can therefore be recommended as a screening procedure in patients with clinical symptoms of disorders in steroid production and/or metabolism. PMID- 3041102 TI - Comparative O2-. responses of lung macrophages and blood phagocytic cells in the rat. Possible relevance to IgA immune complex induced lung injury. AB - IgA immune complex-induced lung injury in the rat is oxygen radical mediated and partially complement-dependent but develops fully after neutrophil depletion. The extent to which monocytes, lung interstitial macrophages, and alveolar macrophages may be involved in the development of lung injury in this model is unclear. To further understand the pathogenesis of IgA lung injury, we have examined the capacity of phagocytic cells isolated from different anatomic compartments of the lung to produce toxic oxygen-derived metabolites. [3H]Thymidine pulse labeling and autoradiography as well as in vivo phagocytosis studies were used to distinguish macrophages isolated from the alveolar and interstitial compartments. Lung interstitial macrophages were characterized ultrastructurally, cytochemically, and functionally. Interstitial macrophages were relatively uniform in size, had blunt pseudopodia, and contained almost no intracytoplasmic lamellar inclusions compared to alveolar macrophages. Similar to monocytes and alveolar macrophages, interstitial macrophages contained nonspecific esterase activity and exhibited the capacity to phagocytize latex and opsonized zymosan particles. Lung interstitial and alveolar macrophages incubated with IgA immune complexes, IgG immune complexes, or phorbol ester (PMA) produced similar amounts of O2-. in a dose-dependent manner. In contrast, peripheral blood neutrophils responded to IgG immune complexes and PMA but not to IgA immune complexes. Monocytes produced a small amount of O2-. in response to PMA but almost no O2-. in response to IgA or IgG immune complexes. These data are consistent with recent in vivo studies which indicate that IgA immune complex lung injury is neutrophil independent. The data provide direct in vitro evidence that lung interstitial and alveolar macrophages produce O2-. following incubation with PMA, IgA, or IgG immune complexes and may therefore contribute to the development of oxygen radical mediated lung injury. PMID- 3041103 TI - A simple enzymic digestion procedure of intact tissue samples in pharmacokinetic drug analysis. AB - A simple procedure is described for the enzymic digestion of intact solid tissue samples. The samples are digested in an enzyme suspension containing collagenase and subtilisin Carlsberg without additional grinding or homogenizing. Bacterial growth during digestion is reduced by addition of an antibiotic solution. The resulting digests contain only very small tissue fragments. The total digestion is carried out at physiological conditions of temperature and pH, which makes the procedure suitable for inclusion in extraction procedures in a broad pH range. Since a minimum of sample manipulation is required, the procedure can be appropriately included in pharmacokinetic studies in which large series of samples are involved. PMID- 3041104 TI - Surface receptor mobilization in complement-mediated neutrophil activation: characterization and effects of methylprednisolone. AB - Complement-mediated neutrophil activation (CMNA) is an important host defense mechanism that is essential for effective neutrophil (PMN) proinflammatory activity. It has also been implicated as a pathogenic mechanism contributing toward the development of adult respiratory distress syndrome. Utilizing zymosan activated serum pretreatment as an in vitro model for CMNA, we characterized the effects of CMNA on PMN superoxide (SO) production and N-formyl-methionyl-leucyl phenylalanine (FMLP) receptor status. CMNA was associated with a 132 +/- 38% increase in FMLP-induced SO generation and a 110 +/- 30% increase in FMLP receptor expression. Methylprednisolone pretreatment prevented both the FMLP receptor mobilization and the SO priming effects of CMNA. These data support a concept that FMLP receptor mobilization is an important element in the PMN activation process. In addition, blocking this phenomenon may have clinical significance in attempts to modulate the potential damaging effects of the increased PMN oxidative metabolism associated with CMNA. PMID- 3041106 TI - The effects of in vivo antibiotics on neutrophil (PMN) activity in rabbits with peritonitis. AB - Antibiotics play an important role in helping the host fight infection; however, the direct cellular effect of antibiotics on polymorphonuclear cells remains undefined. Adherence, chemotaxis, phagocytosis, and superoxide anion production are important steps in the cascade of events initiated by the polymorphonucleocyte in bacterial killing. Previous studies have shown inhibition as well as stimulation of neutrophil antibacterial therapy by antibiotics. Peritoneal and blood polymorphonuclear neutrophils (PMN) respond differently to peritonitis and to external agents. The purpose of this study was to investigate the effects of in vivo clindamycin and netilmicin on infected rabbit peritoneal and blood polymorphonuclear adhesiveness, phagocytosis, chemotaxis, and superoxide anion production. Peritoneal and blood PMNs were obtained from rabbits which had undergone appendiceal devascularization 18 hr earlier: antibiotics were administered intramuscularly 1 hr prior to appendectomy and every 8 hr postoperatively for 5 days; these PMNs were compared to infected rabbits which did not receive antibiotics. Clindamycin and netilmicin in vivo cause significant inhibition of phagocytosis, peritoneal adhesiveness, and, when used in combination, blood adhesiveness and peritoneal superoxide anion production. No effects were seen on chemotaxis. Based on this data we conclude that antibiotics, while vitally important in fighting infections, may in and of themselves be agents of immunosuppression at the cellular level. PMID- 3041105 TI - Influence of inhibitors of ATP catabolism on myocardial recovery after ischemia. AB - The loss of the catabolic products of adenosine triphosphate in the form of purine nucleosides and oxypurines during ischemia and subsequent reperfusion may limit adenine nucleotide regeneration. This study compared the effects of infusion of inhibitors of the major reactions involved in the degradation of adenosine triphosphate to inosine on the postischemic recovery of high energy phosphate and myocardial function. Isolated rat hearts were made totally ischemic after a 5-min infusion of p1,p5-diadenosine pentaphosphate, alpha, beta-methylene adenosine diphosphate, nitrobenzyl-6-thioinosine, or erythro-9-(2-hydroxy-3 nonyl) adenine, which are inhibitors of adenylate kinase, 5'-nucleotidase, adenosine translocase, and adenosine deaminase, respectively. Following 30 min of ischemia, only hearts infused with alpha, beta-methylene adenosine diphosphate recovered significantly better ventricular function than did the control (P less than 0.05), but all hearts had increased adenosine triphosphate and creatine phosphate regeneration (P less than 0.05). The formation and washout of greater than 30% of the total adenine pool metabolites were not prevented by any drug. Nevertheless all manipulations of adenine metabolism resulted in recruitment of high energy phosphate during preischemic infusion which may have potential benefits in elective ischemic arrest. PMID- 3041107 TI - The effect of dexamethasone in vivo on blood and peritoneal neutrophils (PMN) in rabbits with peritonitis. AB - Neutrophils play an essential role in the host's defense against infection. Our previous studies have shown that blood and peritoneal neutrophils (PMN) have different basal activities and responses to infection. We also demonstrated that peritonitis produces divergent changes in the cellular function of PMN both in the blood and in the peritoneal fluid. Steroids are well documented to cause immunosuppression both clinically and, more variably, at the cellular level. Understanding the mechanism of steroid-induced immunosuppression in surgical infection may impart insight on the management of this condition. Using a model of surgical peritonitis, we studied the effects of immunosuppression on rabbit blood and peritoneal PMN. Blood and peritoneal PMNs were harvested after the development of fibrinopurulent peritonitis. Rabbits were divided into two groups: immunosuppressed and control. Immunosuppression was accomplished by intramuscular injection of dexamethasone (2 mg/kg) for 10 days preoperatively and 10 days postoperatively. Purified PMNs were studied for phagocytosis, adhesiveness, superoxide anion production and chemotaxis from both groups. Survival was computed from the number of days the rabbit survived after the operation up to a total of 10 at which time they were sacrificed. Immunosuppression with dexamethasone resulted in inhibition of peritoneal phagocytosis and peritoneal adhesiveness; there were no changes in blood adhesiveness nor blood phagocytosis. Also, there was no significant change in superoxide anion production nor in chemotaxis. Survival of the rabbits was significantly reduced when treated with dexamethasone.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3041108 TI - Synthesis, biologic activity, and protein binding characteristics of a new vitamin D analog, 22-hydroxyvitamin D3. AB - We synthesized a novel vitamin D analog, 22-hydroxyvitamin D3 9 and tested its biologic activity (and antivitamin properties) in vivo in vitamin D-deficient rats, and in vitro in the chick embryonic duodenum. We examined its ability to bind to the sterol carrier protein, vitamin D binding protein and the chick intestinal cytosol receptor for 1,25-dihydroxyvitamin D3. The new vitamin 9 was synthesized from 3 beta-hydroxy-22,23-dinorcholenic acid 1 in 12 steps. The vitamin 9 displayed no vitamin D agonist activity in the intestine or in bone in vivo and did not block the activity of vitamin D3 or 25-hydroxyvitamin D3. It was a weak vitamin D3 agonist in the chick embryonal duodenum in vitro. It did not antagonize the activity of 1,25-dihydroxyvitamin D3. Vitamin 9 bound to the chick intestinal cytosol receptor with low affinity. 22-Hydroxyvitamin D3 and various vitamin D sterols were bound to vitamin D binding protein in the following order: 25-hydroxyvitamin D3. (24R)-24,25-dihydroxyvitamin D3, and (25S)-25,26 dihydroxyvitamin D3 greater than 22-hydroxyvitamin D3 greater than 11 alpha hydroxyvitamin D3 greater than 1,25-dihydroxyvitamin D3 greater than vitamin D3. We conclude that the introduction of a hydroxyl group at C-22 in the side chain of the vitamin D3 molecule decreases its biological activity. PMID- 3041109 TI - Factors influencing production of 5(E)-19-nor-10-keto-vitamin D3 by rumen bacteria. AB - Mixed populations of rumen bacteria or Clostridium hastiforme (a rumen isolate) catalyzed the oxidation of vitamin D3 to 5(E)-19-nor-10-keto-vitamin D3. The reaction depended upon small amounts of O2 (less than 0.1% dissolved O2); when O2 was available, supernatant obtained from heat-killed mixed cultures also produced 5(E)-19-nor-10-keto-vitamin D3. Results obtained by ultrafiltration indicated that at least two heat-stable factors of bacterial origin were involved. Lower rates of the same oxidation were observed when O2 was introduced to solutions containing vitamins D3 and L-cysteine. Oxygen radicals are known to be produced in such solutions and the involvement of such radicals in the D3 oxidation is probable since production in cysteine solutions was inhibited by superoxide dismutase and catalase. PMID- 3041111 TI - The multicomponent analysis of estrogens in urine by ion exchange chromatography and GC-MS--I. Quantitation of estrogens after initial hydrolysis of conjugates. AB - A method for the multicomponent analysis of estrogens in urine after initial hydrolysis of the conjugates is described. Following protection of the carbonyl functions by ethoximation, estrogen conjugates were extracted on Sep-Pak C18 cartridges and purified on the acetate form of DEAE-Sephadex. The samples were subsequently hydrolysed by Helix pomatia juice and the hydrolysate was purified on the acetate form of QAE-Sephadex. Estrogens with vicinal cis-hydroxyls and diphenolic compounds were fractionated on the borate and bicarbonate form of QAE Sephadex, respectively. Neutral steroids were removed by the free base form of DEAE-Sephadex after which estrogens were separated into two groups using Lipidex 5000 in a straight phase system. Following trimethylsilyl ether derivatization estrogens were analysed by selected ion monitoring (SIM). The method allows the quantitation of all the important estrogen metabolites including catechol estrogens. It is precise, accurate and sensitive permitting the quantitation of estrogens in urine of males and non-pregnant females. PMID- 3041110 TI - The effect of ACTH therapy on serum dehydroepiandrosterone, androstenedione, testosterone and 5 alpha-dihydrotestosterone in infants. AB - Serum concentrations of dehydroepiandrosterone (DHEA), androstenedione, testosterone, 5 alpha-dihydrotestosterone and cortisol were measured in 10 infants (age 5-22 months) before, during and after 6-weeks of ACTH therapy for infantile spasms. During therapy, their mean DHEA concentrations increased 2.3 fold, androstenedione 12.3-fold, testosterone 2.7-fold, 5 alpha-dihydrotesterone 2.5-fold and cortisol 2.9-fold compared to pre-therapy values. Serum dehydroepiandrosterone sulphate (DHEA-S) concentrations were also increased during ACTH therapy above the normal prepubertal range. Three days after the cessation of ACTH treatment, all androgens had returned to the pre-therapy level. We conclude: At least in pharmacologic doses ACTH alone stimulates adrenal androgen secretion in infants, excluding the necessity of a separate adrenal androgen stimulating hormone. PMID- 3041112 TI - Proton NMR of human breast tumors: correlation with clinical prognostic parameters. AB - Proton NMR spectroscopy and imaging of human breast tissue have provided new methods in studying breast carcinomas. Continuous wave proton NMR spectroscopy in this study is able to discriminate breast carcinomas from normal breast tissue on the basis of the integrated area under the water and lipid peaks, width at half height of the water peak, and chemical shift of the lipid peak. In addition, the NMR parameters were correlated with the following clinical and pathologic prognostic indices: TNM tumor stage, nuclear grade, and estrogen receptor status (ER). Width at half height of the lipid peak (1/2 delta lipid) correlated with tumor content and ER. Studies using higher resolution proton or phosphorus NMR spectra may separate signals that can correlate with biological information on breast neoplasms useful to the clinician. Chemical shift of the lipid peak may be used to sharpen contrast on MRI of breast tumors. PMID- 3041113 TI - Primary liver cancer: pattern of metastasis. AB - The causes of death and patterns of metastasis of 43 patients with primary liver cancer were studied and discussed in comparison with reports in the literature. Data from 6 recent reports of 1,673 patients showed that the 3 leading causes of death from hepatoma were liver failure (34%), bleeding (30%), and advanced cancer (24%). Distribution of sites of metastasis in 1,497 patients with hepatoma from 7 reports showed that the 3 leading sites were: lung (median 44%), portal vein (35%), and portal lymph node(s) (27%). The pattern of metastasis of hepatoma arising from cirrhotic liver is somewhat different from that of the noncirrhotic liver; the former are more likely to involve the portal vein, whereas the latter involves more regional lymph node(s). Compared to hepatoma, cholangiocarcinoma is less likely to metastasize to the lung or portal vein, but more likely to involve lymph node(s), peritoneum, and bone and/or marrow. PMID- 3041115 TI - Benign mixed tumor (pleomorphic adenoma) of the breast: ultrastructural study and review of the literature. AB - A patient is presented with a benign mixed tumor (pleomorphic adenoma) of the breast. There are 11 well-documented cases of this rare breast neoplasm. It is histologically and ultrastructurally identical to that seen in the salivary gland and follows a similar benign course. A central role of the ductal myoepithelial cell is proposed for the histogenesis of this tumor. PMID- 3041114 TI - Spontaneous penetration of dura mater and bone by glioblastoma multiforme. AB - Three patients are reported whose cranial dura mater and bones were penetrated by intracranial glioblastomas in the absence of previous craniotomy or radiotherapy. The gliomatous nature of the tumors was confirmed by localization of cytoplasmic glial fibrillary acidic protein (GFAP) of the neoplastic cells. Review of the literature disclosed only 15 glioblastomas, including the three cases, spontaneously penetrating the cranial dura mater and bones. These patients ranged in age from 3.5 to 70 years with an average age of 40 years. The male/female ratio was 5/8. Five glioblastomas were in the temporal lobes, three were in the frontal lobes, three were in the frontotemporal regions, two were in the occipital lobes, one was in the frontoparietal region, and one was in the temporoparietooccipital region. Six glioblastomas also had spontaneous distant metastases. In the absence of previous craniotomy and radiotherapy, rapid growth of the glioblastomas may promote such spontaneous penetration into the cranial dura mater and bones. PMID- 3041117 TI - Use of tension measurements to delineate the mode of action of vasodilators. AB - Direct experimental procedures to delineate the modes of inhibition of drugs on vascular smooth muscle contractility are described. In isolated rabbit aortic strips, high concentrations of KCl and norepinephrine induce sustained contractions that depend upon Ca2+ influx mediated by selective activation of voltage-dependent and receptor-linked Ca2+ channels, respectively. In the absence of external Ca2+, norepinephrine also induces a transient contraction that is dependent upon release of cellular bound Ca2+. The contractile responses are differentially susceptible to vasodilators acting on different mechanisms. The Ca2+ channel blockers selectively inhibit KCl-induced contraction. The selective inhibitors of norepinephrine-induced contraction include nitro compounds, atrial natriuretic peptide, acetylcholine and other stimulants of endothelium-derived relaxing factor, beta-adrenergic agonists, forskolin, adenosine, and metabolic inhibitors. The nonselective inhibitors of these contractions include the inhibitors of contractile filaments, such as calmodulin inhibitors, and the inhibitors with multiple sites of action, such as papaverine. Although the inhibitors of Ca2+ release from storage site, such as ryanodine, may not inhibit these contractions, these inhibitors inhibit the norepinephrine-induced transient contraction in Ca2+-free solution. Thus, primary evaluation (screening) of drugs affecting vascular smooth muscle contraction can be performed by analyzing their effects on contractile responses of isolated rabbit aorta. Furthermore, methods to define more detailed sites of action of drugs are also described. PMID- 3041116 TI - Somatostatinoma: a case report and review of the literature. AB - Somatostatinomas are rare endocrine tumors that were first described in 1977. In addition to the present case report, there have been 31 cases reported in the literature. We have reviewed the literature to integrate the symptoms, physical findings, diagnostic tests, treatment, and length of survival of these patients. Although the symptoms that occurred in the majority of cases were those that are seen in most patients with intra-abdominal neoplasms, symptoms relating to the presence of excess circulating somatostatin--diabetes, maldigestion, and cholelithiasis--were frequently seen. Physical findings and the results of diagnostic tests were usually nonspecific. The majority of the patients underwent radical surgical procedures (Whipple procedure or pancreatic resection). The pancreas was the most frequent site of involvement (21/31 cases), but primaries in the duodenum, ampulla of Vater, cystic duct, and jejunum have been described as well. Metastases were most frequently seen in the liver and lymph nodes. Chemotherapeutic agents were administered to 10 patients, usually as adjuvant therapy, and appear to be useful in treating recurrent and metastatic disease. The one-year survival of these patients is 48%, which is better than that for patients with carcinoma of the pancreas or biliary tree. Therefore, it is important that the diagnosis of somatostinoma be made so that the patient may be treated accordingly and followed by serial somatostatin levels for evidence of metastasis or recurrent disease. PMID- 3041118 TI - An in vivo cumulative dose-response assay of the myocardial beta-adrenoceptor system. AB - This paper describes an in vivo computer-based cumulative dose-response assay of the myocardial beta-adrenoceptor system. The technique involves measuring the ability of isoproterenol, a beta-adrenergic agonist, administered through a jugular catheter to increase heart rate in rats. The computer system monitors heart rate, provides real-time graphics of incoming data and a detailed graphic review of responses following testing, coordinates drug injections by the experimenter, performs a nonlinear line analysis to determine the dose required to produce 50% maximal responding (ED50) following the data review, and generates a diskette and printer report at the completion of testing. Data are presented and discussed on how well this system meets the assumptions underlying the cumulative dose-response methodology. The results of a study on the effect of acute footshock on the myocardial beta-adrenoceptor system as measured by this technique is also presented and discussed. This technique permits chronic studies of an important myocardial receptor system and allows receptor changes to be tracked within individual subjects. PMID- 3041119 TI - Examination of cardiac alpha-adrenoceptors from pharmacological responses and radioligand binding. Comparison of rat and guinea pig tissues. AB - The object of this study was to determine suitable experimental conditions for the pharmacological evaluation of cardiac alpha-adrenoceptors. Atrial and ventricular preparations of the guinea pig and rat were employed, and the alpha adrenoceptor responsiveness was compared with the binding of the alpha adrenoceptor radioligand [3H]prazosin in membranes prepared from these cardiac regions. The experimental variables examined were the pacing frequency, bath temperature, choice of agonist, and cardiac region. In guinea pig atria the optimum alpha-adrenoceptor-mediated positive inotropic response to phenylephrine was at 32 degrees C and a pacing frequency of 1 Hz. A comparison of phenylephrine with methoxamine showed that the former yielded biphasic concentration-response curves in guinea pig left atria; the lower portion was alpha-adrenoceptor mediated and the upper, more substantial portion, was beta mediated. Methoxamine produced monophasic curves due entirely to alpha-adrenoceptor stimulation and was therefore used for comparisons between rat and guinea pig tissues. Of the guinea pig tissues, only the left atrium produced positive inotropic responses. Negative chronotropy was obtained with right atria and negative inotropy with ventricular strips and papillary muscles. The rat tissues all responded with positive responses, the largest maximum being in the left atrium. Binding data showed a larger number of alpha-adrenoceptors in the rat tissues, of which the ventricles had the greatest number. The lack of response of guinea pig ventricular tissues was therefore reflected in the low binding. From this study, the most appropriate model for characterizing cardiac alpha-adrenoceptors is therefore the rat left atria at 32 degrees C and paced at 1 Hz with methoxamine as the agonist. PMID- 3041120 TI - Inhibition of neutrophil myeloperoxidase activity by selected tissues. AB - Myeloperoxidase, a polymorphonuclear leukocyte-specific enzyme, has been used previously to quantify the number of polymorphonuclear leukocytes in tissue. When this method was employed in an attempt to measure polymorphonuclear leukocyte numbers in infected kidneys, it was found that myeloperoxidase could not be demonstrated, although significant numbers of neutrophils were present in the pyelonephritic lesions. Further studies were carried out to determine the effect of other tissues on free and cell-bound exogenous myeloperoxidase. We have shown that while skin had little effect on enzyme levels, liver and spleen totally destroyed myeloperoxidase activity within 30 sec. Cardiac and striated muscle had an intermediate effect. When intact neutrophils were added to fresh cardiac tissue 72.5% myeloperoxidase activity was destroyed during the enzyme solubilization procedure. These findings indicate that the technique can only be used for the quantification of polymorphonuclear leukocytes in selected tissues and that appropriate controls are essential. Previous studies in which myeloperoxidase levels have been used to estimate polymorphonuclear leukocyte numbers in cardiac tissue will need reevaluation. PMID- 3041121 TI - Preparation of single smooth muscle cells from guinea pig taenia coli by combinations of purified collagenase and papain. AB - Procedures for isolation of single smooth muscle cells from taenia coli of guinea pigs have been developed. The preparation was performed with a combination of highly purified collagenase prepared by Amano Pharmaceutical Co. (Japan) and papain obtained from Sigma Chemical Co. (Type III). This combination resulted in very high yield of the single cells (39.2 +/- 4.5 X 10(3) cells/mg tissue wet wt) and less cell debris. In the ordinary procedure, commercially available collagenase preparations contaminated with various peptidases have been used. With these enzyme preparations, however, the yield of single cells was dependent on the batch of the preparations, and a large amount of cell debris was contaminated. Combination of the highly purified collagenase and papain resulted in higher yields constantly. Cells, isolated with these enzymes in a medium consisting of 140 mM KCl, 1.0 mM MgCl2, 4.2 mM Hepes, and 5.6 mM glucose (pH 7.4), were spindle shaped. The length of the cells was 185.9 +/- 5.2 micron (n = 90) and the diameter was approximately 12.6 micron. The diameter was not dependent on the cell length. More than 80% of the single cells were viable when examined by trypan blue exclusion technique. Under the depolarized condition, cells remained viable longer because of lower energy consumption, and these cells were contracted by Ca dose dependently. The dose-response relationship was similar to that obtained with intact tissue. Because the cells are constantly available with higher yield, the preparation might be applicable for biochemical research such as ion flux. Details of cell properties under the physiological conditions are under investigation. PMID- 3041122 TI - Effect of gossypol acetic acid on the epididymis: histochemical and scanning electron microscope studies. AB - Rats were treated orally with gossypol acetic acid at 30 or 10 mg/kg daily, 6 days a week, for 8, 12, 14 or 16 weeks. At the end of each treatment regimen, treated rats and an equal number of control rats were killed for histological and histochemical studies. From 8 weeks onward, as a result of the treatment, the tubular lumen of the corpus epididymides became narrowed with thickened pseudostratified epithelium and there was a reduction in the amount of spermatozoa. There was an increase in esterase, alkaline phosphatase, acid phosphatase and ATPase activity. These changes increased in intensity with the duration of treatment. Scanning electron microscopic examinations of the corpus epididymides of rats treated for 16 weeks, compared with those of controls, revealed similar changes, namely, narrowing of the tubular lumen, thickening of the pseudostratified epithelium and reduction in the number of spermatozoa. PMID- 3041123 TI - Effects of gossypol acetic acid on rat luteal cells in vitro. AB - The action of gossypol acetic acid (GAA) on 125I-hCG binding, gonadotropin stimulated cAMP accumulation and progesterone production was investigated utilizing rat ovaries. Incubation of luteal cells for 3 h with increasing concentration of GAA caused a significant inhibition of gonadotropin-stimulated steroidogenesis. The inhibitory effect of GAA was concentration dependent. GAA at concentrations of 10-30 micrograms/ml reduced cAMP formation in response to hCG. It was shown that the activity of adenylate cyclase of luteal cells was inhibited by 10 micrograms/ml GAA. GAA at a concentration of 30 micrograms/ml was found to have an inhibitory effect on 8Br-cAMP-stimulated progesterone production. GAA did not affect 125I-hCG binding to LH receptor on the luteal cell surface. These results suggest that in luteal cells GAA inhibits steroidogenesis at the step of gonadotropin-stimulated cAMP formation. Adenylate cyclase of luteal cells was inhibited. PMID- 3041124 TI - Postoperative T1 N0 non-small cell lung cancer. Squamous versus nonsquamous recurrences. The Lung Cancer Study Group. AB - The disease-free, postresection, 2 year survival rate of patients with T1 N0 non small cell lung cancer surgically/pathologically staged by the Lung Cancer Study Group is about 82%. This study of the rate of cancer recurrence and histopathologic types is based on 572 eligible patients who submitted to complete resection of T1 N0 lung cancer and rigorous, systematic mediastinal lymph node sampling. The initial pathologic interpretation and staging were reviewed by pathologists of the Lung Cancer Study Group Pathology Reference Center to assure uniformity. Review was completed for 82% of patients included in this report. Postoperative cancer recurrence was observed in 107 of the 572 eligible patients. Histopathologic classification was squamous carcinoma (226 patients) or nonsquamous, non-small cell carcinoma (346 patients). Cancer recurrences are more frequent and recurrence rates are higher in the patients with nonsquamous cancer. There is a greater probability of first recurrence in the brain in the nonsquamous carcinoma group. This study substantiates the observation that lung cancer recurrences are histopathologically dependent in the T1 N0 subgroup of Stage I non-small cell lung cancer, with higher rates occurring among patients with nonsquamous carcinoma. PMID- 3041125 TI - Malignant fibrous histiocytoma of left atrium. AB - A case is presented of malignant fibrous histiocytoma arising adjacent to a mitral Carpentier-Edwards bioprosthesis placed 6 years previously. PMID- 3041126 TI - Intracellular cAMP regulates the cytotoxicity of recombinant tumor necrosis factor for L cells in vitro. AB - Tumor necrosis factor (TNF) destroys certain tumor cells in vitro. Using the classical TNF-sensitive cell line L929 and its resistant derivative, we show here that the sensitivity of cells to TNF depends in large measure on the intracellular cAMP concentration. An experimental increase of the intra-cellular cAMP concentration can substantially enhance the sensitivity of L-cells to TNF. This increase in TNF sensitivity is seen only in TNF-sensitive cells but not in TNF-resistant cells. An increase in intracellular cAMP levels has been obtained by adding cell permeable dibutyryl cAMP to the culture medium or by culturing L929 cells in the presence of reagents which cause the elevation of intracellular cAMP concentrations. PMID- 3041127 TI - Beta-adrenergic receptor properties of a pulmonary alveolar type II cell preparation from the adult rat. AB - The pulmonary alveolar type II cell synthesizes and secretes phosphatidylcholine (PC), a major component of surfactant, above basal level in response to beta adrenergic stimulation. The investigation of the specific receptor which mediates these events was the topic of this study. Freshly isolated type II cells from adult rats were disrupted in a French pressure cell, and crude particulate fractions were recovered and used in assays for binding of the radioligand (-)-3 [125I]-iodocyanopindolol. The receptor had high affinity for beta-adrenergic agents, and specific binding to the receptor was saturable and reversible. The KD value obtained by kinetic means (19.6 pM) was in close agreement with that obtained by Scatchard (21.5 pM) and Hill (21.3 pM) analyses of steady-state binding data. The Scatchard correlation coefficients and Hill plot coefficients were close to 1, indicative of a single class of binding sites which displays no cooperativity. The specificity for catecholamine agonists and stereoselectivity observed were appropriate for a beta-adrenergic receptor. Use of selective drugs identified the presence of both beta 1- and beta 2-adrenergic receptor subtypes (1:3, respectively) on this cell type. PMID- 3041128 TI - Relaxant effects of forskolin on guinea pig tracheal smooth muscle. AB - We investigated the relaxant effects of forskolin, a diterpene derivative isolated from the roots of Coleus forskohlii, on guinea pig airway smooth muscle by measuring the isometric tension of tracheal smooth muscle in vitro and transcutaneous Po2 during the histamine inhalation test (HIT) in vivo. Forskolin (10(-9)-10(-5) M) caused dose-dependent relaxant effects on resting tone and on leukotriene C4 (10(-7) M)-, leukotriene D4 (10(-7) M)-, and carbachol (3 X 10(-6) M)-induced contraction of tracheal smooth muscle. Moreover, with propranolol pretreatment the relaxant effect of forskolin on tracheal smooth muscle did not change, whereas with the same pretreatment the relaxant effect of isoproterenol diminished. Forskolin (10(-8)-10(-6) M) raised tissue cyclic AMP levels dose dependently in tracheal smooth muscle (6.7-359.9 pmol/mg protein). Forskolin (1 mg/kg) administered subcutaneously raised the respiratory threshold of (RT histamine in the HIT. The determination of the RT-histamine by measuring tcPo2 was possible without anesthesia. These results suggest that forskolin relaxes airway smooth muscle in guinea pigs in vitro and in vivo by raising tissue cyclic AMP levels and that its actions are independent of beta-adrenoceptors. PMID- 3041129 TI - Cell surface oligosaccharide modulation during differentiation. II. Membrane mobility of oligosaccharide lectin conjugates. AB - The quantitative, population doubling level (PDL) dependent changes in cell surface oligosaccharides on IMR-90 cells, were investigated from the perspective of membrane mobility of the lectin-oligosaccharide conjugates. Concanavalin-A (CON-A), wheat germ agglutinin (WGA), Ricinus communis agglutinin (RCA-120), and Dolichos biflorus agglutinin (DBA) were all observed to cluster, cap, and endocytose in cultured human diploid fibroblasts (IMR-90). Quantitative photometry at 37 degrees C over defined periods of time indicated that as the IMR 90s approached cellular senescence a specific lectin-dependent inability to either endocytose or process the capped complex occurred. The development of a biotin/avidin/enzyme amplification assay permitted the assignment of the accumulating signal to the internal compartment. Kinetic data indicate that there are at least three separate (and separable) mechanisms for the PDL related changes in lectin binding. Data for the CON-A complex indicates that at least two classes of functional complexes are present. Regression analysis of the kinetic data for the RCA-120 complex indicates a similar membrane clearance for the IMR 90s at all population doubling levels (PDL), suggesting that the quantitative differences observed earlier were due to simple quantitative reductions in the RCA complexing molecules. Data for WGA mobility on the membrane indicates that they are both changes in the number and mobility status of the complexes. These results indicate that the quantitative changes in lectin binding observed previously as IMR-90 cells approach senescence are correlated with alterations in membrane mobility patterns of the lectin oligosaccharide conjugates. PMID- 3041130 TI - Control of low Km cyclic adenosine 3',5'-monophosphate-phosphodiesterase in prostate and heart of adult and aged rats. AB - The activity of low Km cAMP-phosphodiesterase (PDE) was determined in extracts of prostate and heart of adult (10-12 months old) and aged (32-35 months old) Sprague-Dawley rats; the enzyme's response to endogenous inhibitors extracted from the two organs was analyzed by kinetic studies. Different mechanisms of inhibition, non-competitive in prostate and competitive in heart extracts, indicate organ-specificity of the inhibitor. The decreased PDE activity in organ extracts of aged rats and its continued sensitivity to the endogenous inhibitor suggest an age-associated impairment of the tissues' ability to terminate cAMP mediated signals. PMID- 3041131 TI - [Dermatomyositis and hepatic carcinoma]. PMID- 3041132 TI - Angiotensin-converting enzyme inhibitors. AB - In summary, ACE inhibitors are effective in reducing blood pressure as initial therapy in some hypertensive patients and in combination with diuretics and other agents in virtually all hypertensives. ACE inhibitors are uniquely advantageous because of their favorable hemodynamic effects, the lack of adverse metabolic effects, and their ability to prevent or blunt undesirable effects of diuretic therapy. Their safety in large numbers of hypertensives has been consistently demonstrated. The minor nature of most side effects and the rarity of life threatening side effects of ACE inhibitors is reassuring. Clinical experience has provided information about patients likely to be at high risk for side effects with ACE inhibitors enabling avoidance of the drugs, or the use of small doses and careful scrutiny in such individuals. The development of this new class of drugs permits safe and effective blood pressure control with potential enhancement of the sense of well being and quality of life to a degree never before encountered. PMID- 3041133 TI - Autoreceptors in the central nervous system. PMID- 3041134 TI - Stereochemistry in the analysis of drug-action. Part II. PMID- 3041135 TI - [Demonstration and organizational structure of the DNA of human papillomaviruses in laryngeal and hypopharyngeal carcinomas]. AB - Thirty biopsy specimens from various histological types of human carcinomas of the larynx and hypopharynx were analysed for the presence of human papillomavirus (HPV) DNA: DNA from the individual specimens were tested for the presence of homologous sequences to HPV genotypes 1, 2, 4, 8, 9, 10, 11, 13, 16 and 18. One squamous cell carcinoma of the hypopharynx (postcricoideal area) contained multiple copies of DNA hybridizing under stringent conditions with HPV 16 DNA. The latter DNA has been found to be frequently associated with human genital cancer. HPV 16 DNA was found mostly episomally as oligomeric circles of 7.9 kbp size, and as larger rear-ranged circular molecules. Integration of the viral DNA in the host cell DNA seems quite likely. Integration and rearrangement of viral DNA into cellular DNA may play a role in the induction and maintenance of the transformed state. The presence of sequences reacting under semistringent conditions with HPV DNA was observed in two additional biopsy specimens of this study. This could suggest that additional laryngeal cancers are associated with papilloma virus infections. PMID- 3041136 TI - [Myeloblastic myoma of the tongue]. AB - Usually the myeloblastoma of the tongue (Abrikossoff's tumour) is a benign tumour. Two cases are reported that differ from those described in the literature in respect of findings, course and histology. Particular reference is made to the malignant nature of this tumour. PMID- 3041137 TI - Carboxypeptidase N (kininase I) activity in blood and synovial fluid from patients with arthritis. AB - Carboxypeptidase N (CPN, kininase I) and kininase II (angiotensin converting enzyme) activities were measured simultaneously in blood plasma and synovial fluid in patients suffering from rheumatoid arthritis (RA), psoriatic arthritis (PA) and osteoarthritis (OA) and in the plasma of normal volunteers. CPN levels (defined as the rate of hydrolysis of furylacryloyl-Ala-Lys) in blood were modestly increased and correlated with erythrocyte sedimentation rate in RA and PA. Based on the hydrolysis of synthetic substrates, CPN activity was much higher than kininase II activity in synovial fluid (SF). SF kininase activities were always inferior to the blood levels in all patients and were correlated with the logarithm of SF leukocyte counts, an indicator of the intensity of inflammation. In addition, CPN and albumin levels in SF were highly correlated when expressed as a percent of the plasma concentrations. Biochemical properties of CPN in crude SF confirmed its similarity to blood CPN. Polymorphonuclear leukocytes derived from inflammatory SF did not release CPN. It is concluded that kininases diffuse from the blood into SF through increased vascular permeability and that CPN could be a major metabolic pathway for kinins in this form of exudate. CPN leads to the formation of des-Arg kinins, selective agonists of the B1 receptors for kinins. PMID- 3041138 TI - Insulin receptor tyrosine kinase activity is unaltered in ob/ob and db/db mouse skeletal muscle membranes. AB - Insulin binding and insulin receptor tyrosine kinase activity were examined in two rodent models with genetic insulin resistance using partially-purified skeletal muscle membrane preparations. Insulin binding activity was decreased about 50% in both 12-week (219 +/- 184 vs 1255 +/- 158 fmoles/mg, p less than 0.01) and 24-week old (2120 +/- 60 vs 1081 +/- 60 fmoles/mg, p less than 0.01) ob/ob mice. In contrast, insulin binding to membrane derived from 24-week old db/db mice was not significantly different from lean controls (1371 +/- 212 vs 1253 +/- 247 fmoles/mg). Insulin-associated tyrosine kinase activity of membranes from ob/ob skeletal muscle was decreased, compared to its normal lean littermate, when compared on a per mg of protein basis in both 12-week (37 +/- 3 vs 21 +/- 3 pmoles/min/mg, p less than 0.05) and 24-week old (71 +/- 5 vs 37 +/- 6 pmoles/min/mg, p less than 0.01) mice. However, no significant differences in kinase activities were observed when the data were normalized and compared on a per fmole of insulin-binding activity basis for the 12-week (12 +/- 1 vs 11 +/- 2) and 24-week (27 +/- 2 vs 20 +/- 3) age groups. Insulin receptor tyrosine kinase activity of db/db skeletal muscle membranes was not different than its normal lean littermate whether expressed on a protein (34 +/- 7 vs 30 +/- 3) or fmole of insulin-binding activity (21 +/- 4 vs 18 +/- 4) basis. These data suggest that insulin receptor tyrosine kinase is not associated with the insulin resistance observed in ob/ob and db/db mice and demonstrate differences in receptor regulation between both animal models. PMID- 3041139 TI - Microinjection of oxytocin into limbic-mesolimbic brain structures disrupts heroin self-administration behavior: a receptor-mediated event? AB - The systemic injection of oxytocin (OXT) decreases the self-administration of heroin in heroin-tolerant rats. Since OXT-ergic binding sites are present in limbic and mesolimbic brain regions, the effects of intracerebral microinjections of OXT were investigated. In heroin-tolerant rats, the microinjection of OXT (2 ng) into the anterodorsal part of the nucleus accumbens or into the ventral hippocampus disrupted the self-administration of heroin. The effect of intrahippocampal microinjections lasted longer than that of intraaccumbens injections. The administration of N alpha-acetyl-(2-0-methyltyrosine)-oxytocin (ACME-OXT), an inhibitor of oxytocin receptors, prevented the disruptive effect of intrahippocampal OXT injections on heroin self-administration. It is concluded that limbic-mesolimbic brain structures have an essential role in the expression of the disruptive action of OXT on heroin self-administration. It appears that OXT-ergic binding sites mediate the effects of OXT. PMID- 3041140 TI - Characterization of GABAA receptor-mediated 36chloride uptake in rat brain synaptoneurosomes. AB - gamma-Aminobutyric acid (GABA) receptor-mediated 36chloride (36Cl-) uptake was measured in synaptoneurosomes from rat brain. GABA and GABA agonists stimulated 36Cl- uptake in a concentration-dependent manner with the following order of potency: Muscimol greater than GABA greater than piperidine-4-sulfonic acid (P4S)greater than 4,5,6,7-tetrahydroisoxazolo-[5,4-c]pyridin-3-ol (THIP) = 3 aminopropanesulfonic acid (3APS) much greater than taurine. Both P4S and 3APS behaved as partial agonists, while the GABAB agonist, baclofen, was ineffective. The response to muscimol was inhibited by bicuculline and picrotoxin in a mixed competitive/non-competitive manner. Other inhibitors of GABA receptor-opened channels or non-neuronal anion channels such as penicillin, picrate, furosemide and disulfonic acid stilbenes also inhibited the response to muscimol. A regional variation in muscimol-stimulated 36Cl- uptake was observed; the largest responses were observed in the cerebral cortex, cerebellum and hippocampus, moderate responses were obtained in the striatum and hypothalamus and the smallest response was observed in the pons-medulla. GABA receptor-mediated 36Cl- uptake was also dependent on the anion present in the media. The muscimol response varied in media containing the following anions: Br- greater than Cl- greater than or equal to NO3- greater than I- greater than or equal to SCN- much greater than C3H5OO- greater than or equal to ClO4- greater than F-, consistent with the relative anion permeability through GABA receptor-gated anion channels and the enhancement of convulsant binding to the GABA receptor-gated Cl- channel. PMID- 3041141 TI - Reversal of the effects of centrally-administered morphine on colonic motility in dogs by the benzodiazepine receptor antagonist RO 15-1788. AB - The effects of intravenous (i.v.) and intracerebroventricular (i.c.v.) administration of morphine on jejunal and colonic motility were investigated in conscious dogs chronically prepared with strain gage transducers and compared to those of i.c.v. DAGO, a highly selective opiate mu agonist. Morphine i.v. (100 micrograms/kg) and i.c.v. (10 micrograms/kg) administered 3 hrs after a meal stimulated colonic motility for 3-5 hrs and induced a phase 3 on the jejunum, which appeared after a 15-60 min delay following i.c.v. administration. These effects were reproduced by DAGO administration at doses of 2 micrograms/kg i.v. and 0.2 micrograms/kg i.c.v. The effects of i.v., but not those of i.c.v., morphine and DAGO on jejunal and colonic motility were blocked by a previous administration of naloxone (100 micrograms/kg i.v.). The colonic stimulation but not the jejunal phase 3 induced by i.c.v. morphine and DAGO were blocked by RO 15 1788 (1 mg/kg i.v.), a selective benzodiazepine antagonist. The colonic stimulation induced by i.v. morphine or DAGO was not modify by i.v. RO 15-1788. It is concluded that i.c.v. administration of mu agonist involved benzodiazepine but not opiate receptors to stimulate colonic motility in dogs. PMID- 3041142 TI - Enhanced binding of morphine and nalorphine to opioid delta receptor by glucuronate and sulfate conjugations at the 6-position. AB - Effect of the modification of morphine and nalorphine by glucuronate and sulfate conjugations at the 3- and 6-positions on the binding to opioid receptors was examined in a particulate fraction of rat brain. Competing potencies of both drugs against [3H]morphine and [3H]leucine enkephalin bindings were extremely decreased by either glucuronate or sulfate conjugation at the 3-position. On the other hand, the potencies of morphine and nalorphine against [3H]leucine enkephalin binding were considerably enhanced by the conjugations at the 6 position, whereas the potencies against [3H]morphine binding were decreased. These altered interactions of the conjugates at the 6-position with the two ligands were attributed to their enhanced binding to delta-receptor and reduced binding to mu-receptor by Hill plot and modified Scatchard analysis. Resulted comparable and simultaneous interactions with mu- and delta- receptors were assumed to be a cause of the enhanced mu-receptor-directed analgesia of morphine and elevated same receptor-directed antagonistic effect of nalorphine, which have been found previously in our laboratory. PMID- 3041144 TI - The benzodiazepine receptor inverse agonists FG 7142 and RO 15-4513 both reverse some of the behavioral effects of ethanol in a holeboard test. AB - The intrinsic effect of the benzodiazepine receptor inverse agonists RO 15-4513 and FG 7142 on the behavior of mice in a holeboard were investigated. Both drugs caused dose-related decreases in exploratory head-dipping. The highest dose of FG 7142 (40 mg/kg) also reduced locomotor activity. RO 15-4513 (1.5 and 3.0 mg/kg) and FG 7142 (10 and 20 mg/kg) reversed the reductions in the number of head-dips caused by ethanol (2 g/kg). The higher doses of these two drugs also partially reversed the locomotor stimulant action of ethanol. Animals that received ethanol in combination with either inverse agonist spent less time head-dipping than vehicle-treated controls. These data indicate that FG 7142 and RO 15-4513 can reverse, at least in part, some of the behavioral effects of ethanol. Neither drug significantly altered blood alcohol concentrations suggesting that the antagonism does not result from pharmacokinetic changes. PMID- 3041143 TI - Modulation of human neuroblastoma transplanted into nude mice by endogenous opioid systems. AB - The role of endogenous opioid systems (endogenous opioids and opioid receptors) in human cancer was explored using an opioid antagonist paradigm and neuroblastoma cells (SK-N-MC) transplanted into nude mice. Mice inoculated with 2.5 X 10(6) neuroblastoma cells received daily injections of either 0.1 or 10 mg/kg naltrexone (=0.1 and 10 NTX groups) which blocked the opioid receptor for 6 8 hr/day or the entire 24 hr/day, respectively, or sterile water. The latency for appearance of a measurable tumor (5 mm diameter) in the 0.1 NTX group was 27% longer than controls (11 days), and the first death in this group occurred 33% later than controls (day 27). Mice inoculated with tumor cells in the 10 NTX group had an acceleration (18%) in the latency of tumor appearance and, 2 weeks after cell inoculation, 70% of the mice in this group had tumors, in contrast to 10% of the controls. At the termination of the experiment (day 45), only 33% of the 10 NTX group were alive, in contrast to 90% of the controls. Receptor binding assays using DAGO, DADLE, or EKC revealed specific saturable binding only for DADLE and EKC. NTX administration resulted in a 148-186% increase in density for both binding sites, but no changes in binding affinity. Measures of opioid levels showed that tumor tissue levels of both beta-endorphin and methionine-enkephalin were elevated 2.5 to 6.5 fold from control values in both NTX groups, whereas plasma beta-endorphin was subnormal by 4 to 6 fold. These results indicate that endogenous opioid systems regulate human neuro-oncogenesis, with opioids being active inhibitors of growth. Opioid antagonists up-regulate receptors and increase tissue levels of endogenous opioids and, under conditions in which the opioid antagonist is short-acting (e.g., 0.1 NTX), can have an exaggerated antitumor effect during the interval when the antagonist is no longer present. PMID- 3041145 TI - Evidence for the direct intervention of angiotensin II in the release of cortisol in teleost fishes. AB - The steroidogenic response to angiotensin II (AII) has been studied in freshwater trout, using a perifusion technique applied to the "head kidneys". AII used alone had no effect on cortisol release. When associated with forskolin or ACTH, it enhanced the stimulation response to these agents. This potentiation was not related (at least directly) to extracellular and intracellular calcium while arachidonate metabolism remained a probable intermediate in the expression of AII action. Experiments using quinacrine and indomethacin suggested that prostaglandin synthesis is involved to mediate AII effect at a step subsequent to cyclic AMP production. These data provide direct evidence that the major components regulating corticosteroid production in teleost fishes are ACTH and AII and that they operate synergistically. PMID- 3041146 TI - Altered tail-skin temperature responsiveness in streptozotocin-induced diabetic rats. AB - We evaluated the tail-skin temperature response to administration of several doses of isoproterenol in streptozotocin-induced diabetic rats after 48 h and after 4 weeks of diabetes. Blood glucose concentrations were significantly increased over controls 48 hours after administration of streptozotocin (65 mg/kg, i.v.) and remained elevated to a similar degree in the 4-week group. Basal rectal temperature and tail-skin temperature (TST) were not different between controls and the diabetic groups and were not affected by administration of saline. However, administration of isoproterenol (25 micrograms/kg, s.c.) caused a significant rise in TST in the control group, but not in the rats diabetic for 4 weeks. A similar but exaggerated response was observed in the controls after subcutaneous administration of 40 micrograms/kg and 100 micrograms/kg of isoproterenol. The TST response in the 4-week diabetic rats still remained negligible with the two higher doses of isoproterenol. When the data were summarized as area under the TST curve, a dose-dependent increase was observed in the control groups and a significant absence of response was observed in the 4 week group. The rats studied 48 h after streptozotocin injection had a similar TST response to the control group after administration of 40 micrograms/kg of isoproterenol. Colonic temperatures did not significantly change between the two groups in any of the studies, although the colonic temperatures tended to rise in the control groups following administration of isoproterenol. We conclude from this study that the absence of a tail-skin temperature response in rats diabetic for 4 weeks results from either a reduced beta-adrenergic receptor mediated response or an altered neural thermoregulatory reflex response, or both. These changes are probably not due to streptozotocin treatment or increases in blood glucose. PMID- 3041147 TI - The effect of muscarinic and beta-adrenergic blockade on cysteamine-induced gastrin secretion by the isolated perfused rat stomach. AB - Cysteamine-induced duodenal ulceration in rats is accompanied by increased circulating gastrin. Although cysteamine appears to exert a direct action on the gastrin cell some groups have provided evidence for an involvement of the autonomic nervous system. The current experiments were performed to determine whether beta-adrenergic or cholinergic (muscarinic) pathways are involved in the acute effect of cysteamine on gastrin secretion in the isolated perfused rat stomach. Cysteamine (1 mM) increased gastrin (IRG) secretion to a maximum ranging between 100% and 192% above basal. A cysteamine concentration of 5mM resulted in peak levels ranging between 150% and 1050% above basal. Addition of atropine or propranalol did not influence the responses obtained. The present results, therefore, do not support a role for either cholinergic or beta-adrenergic pathways in cysteamine-induced gastrin release at the level of the stomach and suggest that in vivo such autonomic effects are mediated extrinsically. PMID- 3041148 TI - Breakdown of phosphoinositides in airway smooth muscle: lack of influence of anti asthmatic drugs. AB - Hydrolysis of membrane inositol phospholipids during agonist-induced contraction in bronchial smooth muscle leads to formation of inositol phosphates. Inositol phosphates are associated with intracellular Ca++ mobilization, which in smooth muscle leads to contraction. We have investigated the effects of inhibitors of the contraction, theophylline, isoproterenol (isoprenaline), and verapamil, on contraction due to carbachol and histamine in bovine airway smooth muscle, and on the formation of inositol phosphates in the same preparation. Since phospholipase C and A2 are involved in the formation of inositol phosphates, we have also studied the effect of inhibitors of phospholipases, dexamethasone and mepacrine, on the accumulation of inositol phosphates. Theophylline, isoproterenol and verapamil elicited a concentration-dependent relaxation of pre-contracted smooth muscle, with the following order of potency: Isoproterenol greater than verapamil greater than theophylline. The relaxant effect was more effective on histamine than on carbachol-induced contraction and depended on the initial airway tone. However, neither theophylline, isoproterenol or verapamil, nor dexamethasone or mepacrine changed the basal level of inositol phosphates or affected the rise due to agonists. We conclude that the smooth muscle effects of theophylline, isoproterenol, verapamil, dexamethasone and mepacrine are not mediated by interference with membrane phosphoinositide breakdown. PMID- 3041149 TI - Effects of GABA on gastric acid secretion and ulcer formation in rats. AB - The effects of gamma-aminobutyric acid (GABA), bicuculline and baclofen, orally and intraperitoneally (i.p.) administered, on the development of stress and pyloric ligation-induced gastric ulcers, were studied in rats. GABA, but not baclofen, significantly reduced the frequency and severity of both models as assessed by ulcer index, incidence and number of ulcers/animal. Gastric protection was dose-related in both experimental models and was completely antagonized by pretreatment with bicuculline methiodide, that blocks peripheral, but not central GABA receptors. All GABA effects were observed after oral and i.p. administration, but inhibition of gastric lesions was greater by the last route. Furthermore, GABA did not affect the acidity or the volume of gastric secretion in pylorus-ligated rats. Consequently its antiulcer activity appears to be mediated by factors unrelated to gastric acid secretion. Since the entry of GABA across blood-brain barrier is greatly restricted it may be concluded that stimulation of peripheral GABA receptors is primarily involved in its antiulcer action. PMID- 3041151 TI - Temperature dependence of proton relaxation times in vitro. AB - Accurate measurement of tissue relaxation characteristics is dependent on many factors, including field strength and temperature. The purpose of this study was to evaluate the relationship between sample temperature, viscosity and proton spin-lattice relaxation time (T1) and spin-spin relaxation time (T2). A review of two basic models of relaxation the simple molecular motion model and the fast exchange two state model is given with reference to their thermal dependencies. The temperature dependence for both T1 and T2 was studied on a 0.15 Tesla whole body magnetic resonance imager. Thirteen samples comprising both simple and complex materials were investigated by using a standard spin-echo (SE) technique and a modified Carr-Purcell-Meiboom-Gill (CPMG) multi-echo sequence. A simple linear relationship between T1 and temperature was observed for all samples over the range of 20 degrees C to 50 degrees C. There is an inverse relationship between viscosity and T1 and T2. A quantity called the temperature dependence coefficient (TDC) is introduced and defined as the percent rate of change of the proton relaxation time referenced to a specific temperature. The large TDC found for T1 values, e.g. 2.37%/degrees C for CuSO4 solutions and 3.59%/degrees C for light vegetable oils at 22 degrees C, indicates that a temperature correction should be made when comparing in-vivo and in-vitro T1 times. The T2 temperature dependence is relatively small. PMID- 3041150 TI - Differences of superoxide production in blood leukocytes stimulated with thymol between human and non-human primates. AB - Thymol induced superoxide production (O2-) by blood leukocytes was examined in various primates including man. Leukocytes of chimpanzee and hamadryas baboon cells showed only 35% of the maximal O2- production rate obtained in human cells, and those of the Japanese monkey and orang-utan failed to respond. In contrast, when cells were stimulated with 12-O-tetradecanoyl phorbol acetate, no significant difference in the O2- production rate was observed between human and monkey cells except for chimpanzee. These results showed that human leukocytes are the most sensitive to thymol among the primates tested. The responsiveness of non-human primate leukocytes could be classified into two types, African type(chimpanzee and baboon) and Asian-type(orang-utan and macaque). PMID- 3041153 TI - [Working Conference on Problems of Proton Therapy. Leningrad, October 14-18, 1986. Proceedings]. PMID- 3041152 TI - Methodology for the measurement and analysis of relaxation times in proton imaging. AB - Measurements of proton T1 and T2 were performed on GdCl3 solutions (20 less than T2 less than 500 msec, 90 less than T1 less than 1000 msec) on large-bore NMR imaging systems operating at 1.0T and 1.5T. CPMG multi-echo (ME), multiple saturation recovery (MSR) and modified fast inversion recovery (MFIR) pulse sequences as well as a sequence that combines and interleaves T1 and T2 weighted data acquisition (which we call "multiple saturation-recovery multiple-echo" (MSRME) were used. The relaxation data are compared to those obtained on a small bore NMR spectrometer operated at 1.5T. T1 and T2 values for the solutions were found to be the same within 10% for the two fields. Reproducibility of measurements of T1, T2 and the unnormalized spin density of the solutions was better than 5%. Systematic errors, amenable to correction through calibration, are noted in the imager T1 and T2 values. T1 and T2 values for some typical neural tissues at 1.5T and body tissue at 1.0T for human volunteers were obtained and are tabulated. PMID- 3041154 TI - [The medical proton complex]. PMID- 3041156 TI - [Proton stereotaxic therapy of cerebral arteriovenous aneurysms]. AB - Stereotaxic proton beam therapy using narrow beams (5-10 mm at 50% of isodose) at the energy of 100 MeV in single irradiation and the delivery of absorbed doses, the maximum dosage being 40-80 Gy, was provided to 99 patients with arteriovenous brain aneurysms. An analysis of the therapeutic results based on angiography data was performed in 50 patients with arteriovenous malformations with the volume up to 10 cm3. A positive effect was obtained in 31 of 50 patients. PMID- 3041158 TI - [3-unit complex for proton therapy]. AB - Since 1969 proton beam therapy of patients with different types of diseases using the ITEP synchrotron proton beam with the energy up to 200 MeV has been conducted in a number of Moscow medical centers. These irradiations employing a specially formed beam are used on a routine basis and are performed in parallel with the program of physical research. Two additional channels as well as two new procedure rooms have been in operation since 1982. A special setup of equipment including clinical dosimetry devices, equipment for patients' irradiation (4 special units), and computer-controlled systems have been installed at the facility. By the present time over 1300 patients have been irradiated. The authors describe physical and dosimetric equipment and irradiation techniques. A summary table containing data on the patients is provided. PMID- 3041155 TI - [Proton therapy: clinico-methodological aspects, treatment results]. AB - In the period of 1975-1986, 457 patients (81 with breast and prostatic cancer, 24 with endocrine ophthalmopathy, 20 with diabetic angioretinopathy, 206 with pituitary adenomas, 115 with arteriovenous and 6 with arterial brain malformations, and 6 with epilepsy) were given proton beam therapy at the Central Research Roentgenoradiology Institute using the 1000 MeV synchrocyclotron of the Leningrad Institute for Nuclear Physics. A prolonged remission was noted in 75 90% of the patients with pituitary adenomas. Angiography showed complete exclusion of aneurysm from the blood flow 2 yrs. after irradiation almost in 50% of the patients with arteriovenous malformations. A high efficacy and safety of the method was shown. PMID- 3041157 TI - [Development of a 2-unit medical complex for proton therapy]. PMID- 3041159 TI - [Proton therapy in clinical neurosurgery]. AB - The use of proton beam irradiation in neurosurgical clinical practice helps to find a solution to the problem of the treatment of inoperable malformations employing this method in some cases as an alternative to surgical intervention. Hypophyseal tumors, tumors of the cavernous sinus, arteriosinusal anastomoses in the cavernous sinus area, and arteriovenous malformations are irradiated at the N. N. Burdenko Institute of Neurosurgery, USSR, AMS using the medical proton beam of the Institute of Theoretical and Experimental Physics. Methods of "piercing" irradiation and methods with the use of Bragg's peak are being developed. At present over 200 patients with hormonally active hypophyseal tumors, 30 patients with tumors of the cavernous sinus, 23 patients with deep seated arteriovenous malformations, and 10 patients with spontaneous arteriosinusal anastomoses in the cavernous sinus area have been irradiated. This method seems to hold promise in neurosurgical practice. Extension of the range of clinical applications of proton beam therapy in neurosurgery requires the development of various techniques of irradiation, all possible approaches to various targets on the basis of contemporary methods of computerized diagnosis, topometry and irradiation design. PMID- 3041160 TI - [Results of treatment of Itsenko-Cushing disease using proton irradiation of the hypophysis]. AB - The application of protons (heavy charged particles) for irradiation of the pituitary body has some essential advantages over other types of teletherapy: insignificant destruction of surrounding tissues, a possibility of a single therapeutic session as well as the absence of post-radiation effects. Proton beam therapy was given to 98 patients with Icenko-Cushing disease aged 15 to 40. Mild cases were treated by proton beam irradiation only while severe cases were managed using proton beam therapy combined with unilateral adrenalectomy or oral administration of ortho-para-DDD. Catamnesis duration varied from 3 to 5 years. In most cases the exposure dose was 80-90 Gy (50-110 Gy). The procedure was well tolerated by all the patients. A dynamic multipolar converting method with 15-20 entrance poles in the left temporal area was employed (with the beam energy of 200 MeV). Stabilization of the course of disease and some clinical improvement were observed in most of the patients 3-4 months after proton beam therapy. In 6 36 months after irradiation 90% of the patients showed normal biochemical indices and the absence of any clinical signs of the disease. Three-five years after treatment, 94% of the patients were in steady remission. A 24-hour ACTH and cortisol normalization pattern was observed in all the patients in parallel with an increase in GH and gonadotropin secretion and a decrease in the prolactin level. Sex function renewal appeared to be the first clinical sign of remission. Young patients aged 15 to 20 with a short duration of disease reached remission faster than patients of the other age groups. Thus the results of proton beam therapy of 98 patients with Icenko-Cushing disease after a follow-up of 3-5 years showed a high efficacy of this type of treatment. The method can be used alone or in combination with unilateral adrenalectomy as well as with oral administration of ortho-para-DDD. PMID- 3041161 TI - [Proton irradiation of the hypophysis and gamma therapy of multiple bone metastases in the complex treatment of breast cancer]. AB - The paper is concerned with the results of multimodality therapy of 190 patients with generalized breast cancer with the prevalence of bone metastasis. Irradiation of the hypophysis by narrow proton beams at a dose of 100-210 Gy and gamma-beam therapy of all affected skeletal zones at a dose of 20-24 Gy in 5-6 fractions were employed in multimodality therapy. The total 50% survival was 42 mos. PMID- 3041162 TI - [Effect of proton irradiation of Tolosa-Hunt syndrome]. AB - The authors have analysed a case of Tolosa-Hunt syndrome with a persistent recurring course and a gradual development of resistance to steroid therapy. Computerized tomography revealed a high density zone in the affected cavernous sinus of the patient. Differential diagnosis between a tumor and Tolosa-Hunt syndrome was made. The patient received 3 fractions of proton beam irradiation by the "piercing" method (the beam being 15 mm in diameter) at a dose of 60 Gy. After irradiation computerized tomography showed complete recovery of disturbed functions and disappearance of the focus. Four-year remission was observed. In our experience, it was the first case of the use of radiation therapy in Tolosa Hunt syndrome. PMID- 3041163 TI - [Proton irradiation of spontaneous arterio-sinus anastomoses in the cavernous sinus region]. AB - Altogether 10 patients with spontaneous arteriosinusal anastomoses in the cavernous sinus area have been irradiated at the N. N. Burdenko Institute of Neurosurgery, USSR AMS. since 1983 using the synchrotron of the Institute of Theoretical and Experimental Physics. A "piercing" method with a proton beam of 12 mm in diameter (in one case 10 mm) was employed. Nine patients had unilateral anastomoses with the blood supply from branches of the internal and/or external carotid arteries, and one patient had a bilateral anastomosis. Irradiation was given in 2 fractions, in 2-3 days, the maximum total dose was 50-60 Gy. Regression of ophthalmological symptoms was noted 2-3 months after irradiation. Convalescence was noted in 8 patients, a follow-up period in 2 patients was insufficient. Of 7 patients examined by angiography complete thrombosis of the anastomosis was noted in 4, considerable reduction of the blood flow was noted in 3. PMID- 3041165 TI - [Initial experience with the use of the proton beam at the Institute of Theoretical and Experimental Physics to treat prostatic cancer]. AB - The proton beam of the 157 MeV synchrotron of the Institute of Theoretical and Experimental Physics was modified for irradiation of prostatic cancer patients. Patient's topometric preparation and method of transperineal irradiation were described. A method of radiotherapy with proton beam boost was applied to 17 patients with different clinical stages of disease. After 6-20 months of the follow-up all the patients have been alive without clinical signs of tumor. In most of the patients radiation reactions of the critical organs were regarded as mild ones. Chronic and long-term radiation injuries were not detected in the patients who lived over 1 year. PMID- 3041164 TI - [Experience in the treatment of eye tumors using a narrow medical proton beam]. AB - This paper presents the effects of proton beam therapy of 175 patients with tumors of the eye. Irradiation was performed with a narrow medical proton beam (Institute of Theoretical and Experimental Physics) with the energy of 70 MeV (Bragg's peak). The total dose was 6000-7000 rad, irradiation frequency was 5-6 sessions every other day. Irradiation was used to treat intraocular and orbital melanomas, melanomas an cancer lesions of the eyelid and conjunctiva. According to the WHO classification, tumors fell under the T2-T3N0M0 stage. The follow-up periods varied from 1 to 10 years (an average of 3.4 years). Complete tumor regression was observed in 45% of the cases, partial regression in 39%, and the absence of the effect in 16%. Twenty-two patients (12%) underwent enucleation due to progressive tumor growth (16 patients) or postradiation complications (6 patients). Hematogenous metastases within a period of 2-5 years following irradiation were noted in 4% of the cases. The results obtained indicated a high efficacy of proton beam therapy for large ocular tumors. The mortality rate from metastases following irradiation was lower than that following enucleation. PMID- 3041166 TI - [Topographic and dosimetric considerations in different methods of proton irradiation]. AB - Two techniques are employed for proton irradiation: one--to pass a proton beam through a target, the other one--to stop in it. The advantage of the first technique is a very small angular divergence of the beam penetrating tissues or, consequently, a very high lateral edge field gradient. The second technique has two additional advantages: the absence of radiation lesions behind the target and an increase in the stopping power (dose) at the end of the beam range localized in the target. The authors present their considerations concerning the applicability of each technique and certain characteristic problems of the second technique. Dose field deformations as a result of topometric uncertainties are also estimated. A method of dose design aimed at minimizing the above mentioned effect is proposed. PMID- 3041167 TI - [Spiral comb filter]. AB - The paper is concerned with some considerations on ridge filters used for proton beam therapy. Such filters are able to transform the beam energy spectrum and the corresponding Bragg curve defining proton beam depth dose distribution. Some difficulties in the manufacture of ridge filters are discussed. The authors describe a ridge filter design which is simple and fit for continuous production. An account of a calculation method and production process is given. PMID- 3041169 TI - [Method of rotation-scanning proton irradiation of esophageal cancer and outlook for its application to the irradiation of tumors at other sites]. PMID- 3041168 TI - [6-unit clinico-physical complex]. AB - The article is devoted to the results of application of high energy proton beams, negative pi-mesons and neutrons in biomedical and clinical investigations carried out at the 680 MeV phasotron (the JINR Laboratory of Nuclear Problems) before its conversion. A six-compartment clinico-physical facility is described. It has been constructed at the converted phasotron in order to continue and expand investigations for application of heavy nuclear particles in radiation therapy. The first physical and dosimetric characteristics of medical proton beams produced in the converted accelerator and guided to chambers 1 and 2 of the new clinico-physical facility are discussed. The future of radiobiological and clinical investigations with high energy proton, negative pion and neutron beams of this facility is also discussed. PMID- 3041170 TI - [Fractionated proton radiotherapy]. AB - Investigations in proton beam therapy of cancer patients have been initiated at the Cyclotron Laboratory, Harvard University, Cambridge, USA, since 1974 using a proton beam with the energy of 160 MeV for fractionated irradiation of uveal melanoma (899 cases), chordoma and chondrosarcoma of the base of the skull (96), sarcoma of the soft tissues and bones (79), prostatic cancer, head and neck tumors, etc. (altogether 1331 patients had been irradiated by June, 1986). To stop a beam in the target computer-assisted three-dimensional design and heterogeneity calculations were performed; computed compensatory boles were produced. Proton beam therapy is used alone or in combination with proton beam irradiation, routine radiotherapy. Areas of particular interest are ocular melanoma, chordoma and chondrosarcoma of the base of the skull, paraspinal sarcomas. Investigations in the field of proton beam therapy of malignant meningioma, metastases to the paraaortic lymph nodes hold promise. PMID- 3041171 TI - [Neurochemical and clinical effects of heterocyclic antidepressants]. PMID- 3041172 TI - [Reproducibility of histologic grading in carcinoma of the breast]. PMID- 3041174 TI - [Molecular cloning of human and animal rotaviruses]. PMID- 3041175 TI - The simian virus 40 large T antigen. A lot packed into a little. AB - Simian virus 40 (SV40) large T antigen is an 81,000 Mr, multifunctional viral phosphoprotein. Certain of its functions are essential to the viral replication process in monkey cells. Others, as yet undefined in biochemical terms, contribute to its neoplastic transforming activity. This review focuses on the known structure-function relationships of this protein and reflects upon the potential significance of its transforming properties in the natural host species of this virus. PMID- 3041173 TI - Molecular cloning of rotavirus genome. PMID- 3041176 TI - Increased leucocyte Na-K ATPase in obesity: reversal following weight loss. AB - Ouabain-sensitive 86Rb influx and [3H] ouabain binding capacity were investigated in the leucocytes of 17 obese patients and 15 control subjects. Both were significantly increased in the obese when compared with controls. Following dietary restriction and a 4% to 5% weight reduction in the obese over 2 weeks, [3H] ouabain binding and ouabain-sensitive 86Rb influx (a model for K+ influx) decreased to levels similar to those in controls. This shows that the number of Na-K ATPase sites on leucocyte membranes of the obese are significantly increased and that this is associated with accelerated 86Rb transport. Since both of these indices decreased following 4% to 5% reduction in body weight while the patients were still obese, increased Na-K ATPase is neither a marker of nor cardinal to the pathogenesis of obesity. We conclude that (1) increase in Na-K ATPase units and 86Rb influx are not characteristic of obesity itself and (2) dietary restriction over the short-term with limited weight reduction restores Na-K ATPase units and 86Rb influx to normal. PMID- 3041177 TI - Catabolism and turnover of collagens: collagenases. PMID- 3041178 TI - Molecular cloning and gene structure of elastins. PMID- 3041179 TI - Isolation and characterization of laminin receptors. PMID- 3041180 TI - Recent developments in posttranslational modification: intracellular processing. PMID- 3041182 TI - A molecular study of Salmonella strains identified from two food-poisoning outbreaks. AB - A molecular epidemiology study was carried out on Salmonella mbandaka and Salmonella corvallis strains identified from two food-poisoning outbreaks which occurred in August 1985 in Pistoia and in October 1985 in Sant'Ilario d'Enza (RE). All the Salmonella mbandaka strains were plasmid-free; all the epidemic Salmonella corvallis strains, in contrast to the non epidemic isolates, carried a small plasmid of approximately 2 MDa molecular weight. Restriction enzyme cleavage pattern analysis revealed that the plasmids of the epidemic strains were closely related. PMID- 3041181 TI - Preliminary characterization of an inhibitory activity of fetal bovine serum on the infectivity of rotavirus strain SA-11. AB - In the present work we studied non antibody inhibiting activity present in fetal bovine serum and active to Rotavirus infectivity and growth in cell cultures. This inhibitor was revealed by an in vitro neutralization test and characterized by gel filtration and chemical and enzymatic treatments. Furthermore, commercial preparations of bovine serum proteins were tested for inhibitory activity. Our results show that serum inhibition is partially resistant to trypsin and neuraminidase treatments but completely destroyed by KIO4. A similar activity was observed in a commercial serum bovine fraction containing predominantly alpha globulins. These results seem to indicate that glycoproteins, and their glucidic components are the molecules predominantly involved in serum inhibition towards Rotavirus infectivity. PMID- 3041184 TI - Comparison of five cell lines for the propagation of bovine viral diarrhea and infectious bovine rhinotracheitis viruses. AB - A comparative study was carried out to determine the susceptibility of 5 different cell types to bovine viral diarrhea (BVD) and infectious bovine rhinotracheitis (IBR) viruses. The cell systems used were swine testicle (ST), mink lung (ML), bovine turbinate (BT), porcine kidney (PK15) and equine dermal (ED) cells. For BVD virus, the titers obtained on day 8 post-infection were 10(1.13), 10(3.25), 10(4.13), 10(0.00) and 10(0.00), in ST, ML, BT, PK15 and ED cells, respectively. For the IBR virus, the titers obtained were 10(2.63), 10(8.5), 10(8.38), 10(3.00), and 10(4.63) in ST, ML, BT, PK15 and ED cells, respectively. This indicates, therefore, that BT and ML cells are optimal for the propagation of both BVD and IBR viruses and that ST, PK15, and ED cells are not very sensitive to these viruses. PMID- 3041183 TI - Antibodies to papillomavirus genus-antigens in women with genital warts. AB - Thirty-three sera from women symptom-free for papillomatous disease and 27 sera from women with flat or exophytic genital warts were examined for the presence of IgG and IgM antibodies to papillomavirus (PV) genus-antigens. For this purpose, sera were challenged with genus-antigens extracted from both human and bovine purified virions of PVs, in a micro solid-phase enzyme-linked immunosorbent assay (ELISA). The assay performed with PV genus-antigens of human origin showed that in women with genital warts, IgG antibodies were present in a percentage of 70.37% and IgM antibodies in a percentage of 40.74%; in apparently uninfected women, IgG and IgM antibodies were present in a percentage of 54.54% and 24.24% respectively. When sera were challenged with PV genus-antigens of bovine origin for IgG antibody class, positivity was 70.37% in women with genital disease and 45.45% in symptom-free women. IgG and IgM antibody response in women with and without papillomatous genital lesions is discussed. PMID- 3041185 TI - Pyridoxine neuropathy. AB - A case of sensory neuropathy in a young woman due to long-term ingestion of pyridoxine, with subsequent recovery, is described. Pyridoxine neuropathy may occur after the long-term ingestion of doses as low as 200 mg a day. Because of its widespread use in the community, both the general public and the medical community need to be aware of this recently described complication of megavitamin therapy. PMID- 3041186 TI - [In vitro effects of potassium superoxide: possible implications for technicians at photography laboratories]. PMID- 3041187 TI - Synthetic diamonds as in vivo radiation detectors. AB - Synthetic diamonds with controlled amounts of impurity atoms can be manufactured so that, as thermoluminescent dosimeters, they can be made to have sensitivities at least as good as presently available commercial thermoluminescent dosimeters. They also exhibit, for radiations normally found in therapy situations, a linearity of response that extends from less than 0.01 Gy (1 rad) to over 10 Gy (1000 rad). Their physical size and form, crystals which can have volumes of less than 1 mm3, make them ideal candidates for in vivo monitoring of radiation fields, particularly electron fields where high-resolution measurements are essential for accurate isodose line determinations. Aspects of dose response from gamma-ray beams in relation to the type and concentrations of the impurity atoms within the diamond are discussed, and some experimental values for gamma, x-ray, and electron beams are presented. PMID- 3041189 TI - Sintered hydroxyapatite for a percutaneous device and its clinical application. AB - The sintered hydroxyapatite was designed to be utilized as a percutaneous device. The device was percutaneously fixed through the back skin of mongrel dogs. The tissue reaction around the implants was examined histologically. At 1 month the sintered hydroxyapatite was closely contacted with the skin tissue and downgrowth of epidermis was not observed. The long-term implantation ranging from 3 to 17 months showed that the depth of epidermis downgrowth was limited to only 1 mm. The amino acid composition of fibrous capsule formed around the hydroxyapatite showed close resemblance to vertebrate periosteum. It was confirmed that the sintered hydroxyapatite had good compatibility and long-term stability with skin tissue. PMID- 3041188 TI - Continuous three-dimensional radiation dosimetry in tissue-equivalent phantoms using electron paramagnetic resonance in L-alpha-alanine. AB - A new tissue-equivalent phantom material has been developed which also acts as a dosimeter. The new phantom material has a similar elemental composition to that of soft tissue and has a density 1.1 g/cm3. The phantom has an agar-gel base, and contains crystallized L-alpha-alanine which traps radiation-induced free radicals. Samples from the phantom were analyzed by an electron paramagnetic resonance (EPR) spectrometer and the intensity of the EPR signal was related to the absorbed dose. When calibrated, the phantom material acts as a dosimeter, with applications in radiation therapy. PMID- 3041190 TI - Effects of hypertonicity on cAMP production in cultured renal epithelial cells (LLC-PK1). AB - The activation of adenylate cyclase by vasopressin in the renal medulla takes place in a hypertonic environment whose osmolality fluctuates widely under varying physiologic conditions. We utilized the cultured renal epithelial cell line, LLC-PK1 as a model system to study the effects of hypertonic electrolyte and nonelectrolyte solutes on the vasopressin-adenylate cyclase interaction. These cells do not produce prostaglandins, thus permitting separate evaluation of the direct effects of hypertonic solutes on the adenylate cyclase response. In intact cells, 40-400 mM NaCl and 600 mM sucrose and mannitol increased basal and vasopressin-sensitive cAMP production 2 to 4-fold. Urea in a concentration range of 300-1,200 mM blunted the stimulatory effect of hypertonic NaCl in intact cells. In order to distinguish direct effects of solutes on the adenylate cyclase response, from effects related to hypertonic cell shrinkage, the influence of these same electrolyte and nonelectrolyte solutes on adenylate cyclase activity in membrane particulate fractions was also examined. Increasing NaCl in the concentration range of 25-100 mM increased basal and vasopressin-sensitive adenylate cyclase. This effect was not specific to sodium, since similar degrees of stimulation were seen with the addition of KCl. Addition of higher concentrations of NaCl, sucrose, and mannitol directly in the adenylate cyclase assay were inhibitory. These findings suggested that the stimulatory effect of hypertonicity in the intact cells was not due to a direct effect of these solutes on the enzyme, but rather to hypertonic cell shrinkage.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3041191 TI - Effects of trimethyltin on granule cells excitability in the in vitro rat dentate gyrus. AB - The effects of trimethyltin (TMT) on passive properties and synaptic activity of dentate granule cell (GC) have been investigated in hippocampal slices in vitro. Intracellular recordings from GC indicated that TMT (1 and 10 microM) increased input resistance from 34.1 +/- 3.6 Mohms to 45.6 +/- 4.1 and 64.7 +/- 14.7 Mohms, respectively, 15 min after its application. This was accompanied by a 10-20 mV depolarization. A decrease in IPSP amplitude was also observed, but developed with longer delays (2-4 hr) following TMT exposure. Extracellular recording from the GC layer during paired pulse stimulation of the perforant path showed a decrease in the ratio of the amplitude of the first to the second population spikes (at an interpulse interval of 9 msec), from 1.8 +/- 0.14 to 0.8 +/- 0.08 (p less than 0.05). The amplitude of the first (conditioning) pulse remained unchanged, suggesting that TMT produced a specific decrease of inhibitory efficacy. These results add evidence to the hypothesis that TMT neurotoxicity is mediated by a decrease of inhibitory synaptic functions. PMID- 3041192 TI - Delta-9-tetrahydrocannabinol during pregnancy in the rat: I. Differential effects on maternal nutrition, embryotoxicity, and growth in the offspring. AB - Either 15 or 50 mg/kg of delta-9-tetrahydrocannabinol (THC) in sesame oil was administered by gastric intubation to gravid rats during the last two weeks of gestation. A pair-fed control group was administered the vehicle alone and allowed to eat and drink only the amount consumed by the 50 mg/kg group on the same gestation days. A nontreated control group was left undisturbed during pregnancy. All treated and control litters were fostered at birth to untreated dams. Among the dams receiving 50 mg/kg of THC, food and water intake was initially reduced to 75-80% of nontreated controls but then recovered over 3-4 days to approximately a 15-20% reduction until term. Compared with the nontreated dams, both THC dose-level groups and pair-fed control dams gained significantly less body weight from conception to term. Offspring mortality did not differ between the nontreated and pair-fed controls but was significantly higher among both dose-level THC exposed groups. In addition, there was a dose-related increase in the sex-ratio of live male to female offspring as well as significant effects on rate of growth for both sexes. The results are discussed with respect to published animal and clinical studies of cannabinoid exposure during pregnancy. PMID- 3041193 TI - Delta-9-tetrahydrocannabinol during pregnancy in the rat: II. Effects on ontogeny of locomotor activity and nipple attachment in the offspring. AB - Either 15 or 50 mg/kg delta-9-tetrahydrocannabinol (THC) in sesame oil was administered by gastric intubation to gravid rats during the last two weeks of gestation. A pair-fed control group was administered the vehicle alone and allowed to eat and drink only the amount consumed by the 50 mg/kg group on the same gestation days. A nontreated control group was left undisturbed during pregnancy. All treated and control litters were fostered at birth to untreated dams. Intact litters from the two THC treated and the two control groups were tested at 3-day intervals from birth to 32 days of age for differences in locomotor activity. In addition, pups were tested for nipple attachment on days 2, 5, 8, 11, and 14 of age. There were no differences in locomotor activity among any of the groups although activity level varied during development. Pups from dams exposed to 50 mg/kg of THC, as well as the pair-fed controls, displayed significantly longer latencies to attach to a nipple. These results suggest that the impaired nipple attachment observed among the high-dose offspring was not a primary effect of THC, but rather was secondary to the significant reduction of food and water intake among the dams. The behavioral findings are discussed with respect to other animal and clinical reports of prenatal cannabinoid exposure. PMID- 3041194 TI - Perinatal diazepam exposure: behavioral and neurochemical consequences. AB - In the present study the effects of perinatal diazepam (DZP) exposure on behavior and benzodiazepine binding site characteristics in offspring were investigated. Pregnant F344 rats were treated during the last trimester of gestation with vehicle or diazepam (3 mg/kg, 5 mg/kg, or 10 mg/kg). Lactating dams were similarly treated on postnatal days 1-7. Both prenatal and postnatal exposure to diazepam resulted in significant effects on the acquisition and extinction of active avoidance behavior measured postweaning. The number of trials to extinction of one-way active avoidance behavior was significantly greater in offspring exposed gestationally to 3 mg/kg and 10 mg/kg diazepam and postnatally to 10 mg/kg diazepam. The mean response latencies for all diazepam treated groups were significantly shorter than those of the vehicle treated rats during the first 15 trials under extinction conditions. In contrast, neither gestational nor postnatal diazepam exposure significantly affected either acquisition or retention of a passive avoidance task. In addition, the binding affinity between the benzodiazepine type I selective ligand, CL218,872, and cortical membranes, as well as the degree to which GABA potentiated 3H-flunitrazepam were significantly altered by perinatal diazepam exposure. No treatment altered the approximate number of benzodiazepine binding sites in either the cortex, hippocampus, or cerebellum. The results of this study further support diazepam as a behavioral teratogen and give new evidence for neurochemical effects following gestational as well as postnatal diazepam exposure. PMID- 3041195 TI - [Correlation between dental pulp pain, blood levels of ACTH, cortisol and beta endorphin and the analgesic efficacy of acetylsalicylic acid]. PMID- 3041196 TI - Histamine receptors in the brain. AB - In mammalian brain, neuronal histamine is likely to act as a neurotransmitter and is recognized by the two classes of histamine receptors (H1 and H2) previously characterized in peripheral organs. Cerebral H1 receptors can be selectively labeled by a tritiated antagonist mepyramine, in particulate fractions or in the living animal. Cerebral H1 receptors mediate the glycogen hydrolysis and the breakdown of inositol phospholipids elicited by the amine. They are indirectly involved in the histamine-mediated accumulation of cyclic AMP. All these biochemical responses mediated by H1 receptors are calcium-dependent. H2 receptors are coupled to an adenylate cyclase. In addition, a novel class of histamine receptors (H3) are presynaptic autoreceptors and modulate the release of neuronal histamine. PMID- 3041197 TI - [Non-invasive tissue oxygen monitoring by near-infrared spectroscopy]. AB - Biological tissues are relatively transparent to light in near infrared region. Brain oxygen metabolism was non-invasively studied by the simultaneous measurement of hemoglobin (Hb) oxygenation and cytochrome oxidase (Cyt. aa3) redox state in the rat brain using near infrared transmission spectrophotometer. Wavelength pairs at 700-805 nm and 830-805 nm were used for the measurements of Hb oxygenation level and of the redox state of cyt. oxidase in situ. Change in the brain blood volume was also monitored by the absorption change at 805 nm. Our estimation of the oxygenation level of brain Hb revealed that present optical measurement picks up mainly that of venous blood. Sharp reduction of the oxidase occurred at the oxygen concentration below 15% in the inspired gas. But above 30% O2, about 85% of the cyt. oxidase was in the oxidized state which was almost constant up to 100% O2. Applications of our techniques for the studies of brain metabolism in acute hemorrhage and in Ca++-antagonist administration were also presented. This non-invasive optical method for the study of human brain metabolism is concluded to be a promissing technique. PMID- 3041198 TI - [Mixed ductal and acinar cell cancer of the pancreas head; report of a case]. AB - A 63 year old Japanese man was admitted in Feb. 1983, with his chief complaint of upper abdominal pain. Physical examination showed only resistance in the right hypochondrium on palpation, but no icteric conjunctiva and skin. A large global tumor of the pancreas head was visualised as a hyperechoic mass with irregularly shaped cystic cavity in ultrasonography, as a hypervascular mass with lucent area in celiac arteriography, and as a mass lesion with low density area in body computerized tomography. Cancer cells were histologically confirmed on specimens taken by fine needle aspiration biopsy under ultrasonic guidance. Cancer of 6.5 X 6.0 X 4.0 cm in size was resected by pancreaticoduodenectomy. Four months after operation, two liver metastatic nodules were resected by right hepatic lobectomy. Histologically, tumor was composed of two characteristic patterns, acinar cell cancer and duct cell cancer, which were confirmed by immunohistochemical techniques. The patient is doing well 3 years and 3 months postoperatively without evidence of recurrent cancer. To our best knowledge, this case is the seventh of mixed ductal and acinar cancer in the world, but the previous 6 cases were reported on autopsy specimens. PMID- 3041199 TI - Increase in the levels of activity of polyadenylic acid-metabolizing enzymes following phytohaemagglutinin stimulation of human lymphocytes. AB - Increased levels of soluble activity of all three enzymes involved in polyadenylic acid metabolism were measured in PHA-stimulated versus normal lymphocytes. Poly(A)-polymerase and poly(A)-exonuclease values increased significantly (from 25.7 +/- 4.2 (S.E.M.) to 53.5 +/- 10.6 (S.E.M.), and from 334.6 +/- 33.2 (S.E.M.) to 653.2 +/- 53.4 (S.E.M.) respectively), while a moderate increase was observed in poly(A)-endonuclease (from 299.2 +/- 33.8 (S.E.M.) to 403.0 +/- 77.1 (S.E.M.). The above differences persisted after two fractionations of the crude cell extracts by ion exchange chromatography and molecular sieving, and could not be attributed to the competitive action of all three enzymes in the untreated extracts. Fractionation of the extracts of resting and stimulated cells on Sephadex G-75 revealed two molecular forms of poly(A) polymerase activity. PMID- 3041200 TI - Long term regulation of glycogen metabolizing enzymes by insulin in H4 hepatoma cells. AB - Insulin alone at concentrations of less than 1 to 5 uU/ml increased the enzyme activities of glycogen synthase, synthase phosphatase, phosphorylase, and phosphorylase phosphatase in hepatoma H4 cells in culture in the presence and absence of serum. Increase in total and active forms of glycogen synthase and phosphorylase were observed. Cycloheximide blocked the action of insulin on glycogen synthase, glycogen synthase phosphatase and phosphorylase phosphatase. The enzymes with the exception of glycogen synthase phosphatase were expressed with greater hormonal sensitivity in the absence as compared to the presence of serum in terms of hormone concentration required and or time of onset. These results demonstrate that these glycogen metabolizing enzymes are under long term control by insulin, with glycogen synthase being the most sensitive of the enzymes studied to the action of the hormone. PMID- 3041201 TI - [Composite transposons Tn5 and Tn10 in Escherichia coli K12: precise excision and recombination]. AB - The number of exconjugants having the transposon Tn5 excised precisely during the crosses of the Escherichia coli proA::Tn5 donor with the recipients F- rec+ or F- recA441 (tif) was 20-30 times higher for the crosses involving the latter recipient. The high recombinogenic activity is characteristic of the tif recipient. Precise excision from a tandem duplication is more efficient than from nonduplicated region of the genome. It is four orders higher, if a transposon is localized in an arm of a duplication. The effect is recA-dependent. The presented data permit us to suggest the participation of RecA protein (its synaptic function) in the formation of the intermediate "stem-loop" structure. The latter is predicted by the three mechanisms of transposon excision: "slippage", "correctional" and "recombinational". The latter two mechanisms were formulated in the paper. The experimental proof of the postexcision transposition presented in the paper, is a good support to the version of "recombinational" excision. PMID- 3041202 TI - [Properties of transposon Tn2555 carrying the genes for sucrose utilization]. AB - Tn2555, a new transposon coding for genes of sucrose utilization was studied. Tn2555 was shown to integrate into the plasmids RP4 and R6K, phage P1CmClr100 and Escherichia coli K12 chromosome. Tn2555 frequency of transposition to RP4 and R6K DNA is (2-5) X 10(-7) in Rec+-strain, (3-6) X 10(-8) in Rec--strain. Tn2555 integration site in phage P1CmClr100 Sac+-derivative studied has been localised within the C-segment of P1 DNA. In three independent cases of Tn2555 transposition to the chromosome the transposon was found to be integrated in the region between 29 and 32 min of Escherichia coli K12 linkage map. The restriction endonuclease analysis of seven independent isolates of RP4::Tn2555 has shown the grouping of Tn2555 integration sites in the Tn1 region of RP4. Frequent rearrangements occurring within Tn2555 have been revealed by the analysis. However, an invertible DNA segment of about 6-7 kb was preserved in all transposon structures. PMID- 3041203 TI - [Expression of human leukocyte interferon A gene in Saccharomyces cerevisiae under the control of regulatory yeast elements PHO5, GAL1 and GAL10]. AB - A chromosomal gene for human leucocyte interferon A is expressed in Saccharomyces cerevisiae yeasts due to interaction of 5'-nontranslating region of the cloned interferon gene with the regulatory elements of yeast genes PHO5, GAL1 and GAL10. Regulated systhesis of interferon was obtained in all cases. The level of interferon genes expression in case using GAL1 and GAL10 genes regulatory elements (5 X 10(5) and 5 X 10(6) u X l-1) correlated with the distances to their promoters. The highest yield of interferon (10(8) u X l-1) was obtained when the PHO5 gene regulatory elements were used. PMID- 3041204 TI - Chromosomal aberrations induced by the restriction endonucleases EcoR I, Pst I, Sal I and Bam HI in CHO cells. AB - 4 widely used cohesive end-producing restriction endonucleases (REs), EcoR I, Pst I, Sal I and Bam HI were tested in CHO cells for their aberration-inducing effects. It was demonstrated that all these REs significantly increased the frequencies of aberrant cells, the aberration frequencies per cell and the aberration frequencies per chromosome. The effects of REs on chromosomal aberrations are similar to ionizing radiation, but more minutes and interchange figures are observed. Polyploid cells are more susceptible to RE treatment, an interesting finding which may be explained by the mechanisms leading to the formation of polyploid cells. PMID- 3041205 TI - RecA-independent mutagenesis in Escherichia coli may be subject to glucose repression. AB - The frameshift mutagen 9-aminoacridine (9AA) causes DNA damage via a recA+ independent mechanism in Escherichia coli. In this study we have exposed E. coli cells carrying the lacZ19124 frameshift marker to 9AA in defined minimal media, washed them, and plated to score for Lac+ revertants. Our results show that 9AA induced reversion to Lac+ occurs in the absence of any exogenous carbon source and when cells are plated on media which do not allow much, if any, cell replication prior to expression of the revertant phenotype. When glycerol (1% w/v) was added to the liquid treatment medium, the number of Lac+ E. coli revertants was similar to that obtained when no carbon source was present. By contrast the addition of glucose (1% w/v) during the mutagenesis treatment caused a significant decrease in the number of revertants. Further experiments indicate that the repressing effects of glucose may be due to a reduction in cAMP concentration, since 9AA mutagenesis was abolished in a cya strain in which no adenylate cyclase is produced. These results are consistent with (but do not prove) the notion that at least one part of the process leading to 9AA mutagenesis is subject to catabolite repression. PMID- 3041206 TI - Repair of benzo[a]pyrene-initiated DNA damage in human cells requires activation of DNA polymerase alpha. AB - Normal human fibroblasts treated with r-7,t-8-dihydroxy-t-9,10-epoxy-7,8,9,10 tetrahydrobenzo[a]pyrene (BPDE) yielded DNA polymerase alpha with elevated levels of activity, incorporated [3H]thymidine as a function of unscheduled DNA synthesis, and exhibited restoration of normal DNA-strand length as a function of unscheduled DNA synthesis. Lipoprotein-deficient fibroblasts treated with BPDE did not show elevated levels of DNA polymerase alpha activity, exhibited minimal [3H]thymidine incorporation, and had fragmented DNA after 24 h of repair in the absence of lipoprotein or phosphatidylinositol supplementation. When DNA polymerase beta activity was inhibited, cells with normal lipoprotein uptake exhibited [3H]thymidine incorporation into BPDE-damaged DNA but did not show an increase in DNA-strand length. DNA polymerase alpha activity and [3H]thymidine incorporation in lipoprotein-deficient fibroblasts increased to normal levels when the cells were permeabilized and low-density lipoproteins or phosphatidylinositol were introduced into the cells. DNA polymerase alpha isolated from normal human fibroblasts, but not from lipoprotein-deficient fibroblasts, showed increased specific activity after the cells were treated with BPDE. When BPDE-treated lipoprotein-deficient fibroblasts were permeabilized and 32P-ATP was introduced into the cells along with lipoproteins, 32P-labeled DNA polymerase alpha with significantly increased specific activity was isolated from the cells. These data suggest that treatment of human fibroblasts with BPDE initiates unscheduled DNA synthesis, as a function of DNA excision repair, which is correlated with increased activity of DNA polymerase alpha, and that increased DNA polymerase alpha activity may be correlated with phosphorylation of the enzyme in a reaction that is stimulated by low-density lipoprotein or by the lipoprotein component, phosphatidylinositol. PMID- 3041207 TI - Somatosensory evoked response evaluation of cervical spondylytic myelopathy. AB - There were 13 patients with cervical spondylytic myelopathy (CSM) evaluated. All had extradural defects with distortion of the cervical cord and partial or complete obstruction of myelographic dye. Posterior tibial scalp (SSEPs) were absent or delayed in all 13, whereas median SSEPs were abnormal in 9. Median SSEP abnormalties were limited to those with cord lesions at C5-C6 or above. There were eight patients with associated radiculopathies confirmed by electromyography. No meaningful differences in SSEPs were noted between those with or without root injury. Surgery confirmed the level of cord injury in 10. CSM is common with a high morbidity. The results indicate SSEPs using leg stimulation can be a sensitive indicator of cord injury in these patients and that the more commonly used arm stimulation is of value primarily for localizing the level of the myelopathy. PMID- 3041208 TI - The neurophysiologic investigation of small fiber neuropathies. AB - We have applied our technique for the measurement of thermal thresholds to 25 patients referred with symptoms and signs of small fiber peripheral neuropathy in whom conventional electrophysiological indices were individually within the range of normal values for our laboratory. Vibration threshold determinations were also within normal range. Significant abnormalities of thermal thresholds were noted in all patients. The results indicate that the technique provides an accurate, easily performed and reproducible index of function in small A delta and C groups of nerve fibers. PMID- 3041209 TI - Hypogravity-induced atrophy of rat soleus and extensor digitorum longus muscles. AB - Prolonged exposure of humans to hypogravity causes weakening of their skeletal muscles. This problem was studied in rats exposed to hypogravity for 7 days aboard Spacelab 3. Hindlimb muscles were harvested 12-16 hours postflight for histochemical, biochemical, and ultrastructural analyses. The majority of the soleus and extensor digitorum longus fibers exhibited simple cell shrinkage. However, approximately 1% of the fibers in flight soleus muscles appeared necrotic. Flight muscle fibers showed increased glycogen, lower subsarcolemmal staining for mitochondrial enzymes, and fewer subsarcolemmal mitochondria. During atrophy, myofibrils were eroded by multiple focal losses of myofilaments; lysosomal autophagy was not evident. Tripeptidylaminopeptidase and calcium activated protease activities of flight soleus fibers were significantly increased, implying a role in myofibril breakdown. Simple fiber atrophy appears to account for muscle weakening during spaceflight, but fiber necrosis is also a contributing factor. PMID- 3041213 TI - Cloning of a Schistosoma japonicum gene encoding a major immunogen recognized by hyperinfected rabbits. AB - A library of randomly sheared Schistosoma japonicum genomic DNA fragments was constructed in the bacteriophage expression vector lambda gt11. A portion of the library was screened with sera collected from rabbits 8 weeks after they were infected with 1000 cercariae. Four clones whose recombinant gene products react with the rabbit sera were purified to homogeneity. Clone SjIR-12A was chosen for detailed study because of its very intense reaction with the rabbit sera. SjIR 12A was found to encode part of a 70 kDa protein (Sj70) that is present in both soluble egg antigen (SEA) and soluble worm antigen preparations (SWAP). Western blot analysis suggests that Sj70 is the only SWAP component that is strongly immunoreactive with the rabbit sera. Rabbit antibodies that react with the SjIR 12A fusion protein were immunoaffinity purified and used to localize immunoreactive product to the nervous tissue of male and female adult worms, the dorsal and lateral tegument of male adult worms, and in eggs to the miracidial tegument and the area between the eggshell and miracidium. Southern hybridization analysis suggests there are approximately four copies of the Sj70 gene per haploid genome. PMID- 3041210 TI - The 5' flanking sequence of a Trypanosoma brucei variable surface glycoprotein gene. AB - The mechanism controlling transcription at several telomeric expression sites for variable surface glycoprotein (VSG) genes in Trypanosoma brucei is unknown. Most VSG genes in expression sites have a region 5' of the gene lacking restriction enzyme sites. This 'barren region' is involved in recombination events which replace the VSG gene with a copy of a different, non-telomeric, VSG gene leading to a switch in VSG expression. Alterations in the barren region have been considered as possible modulators of expression of the adjacent VSG gene in other switching events where no gene replacement occurs. The expressed copy of the ILTat 1.3 VSG gene remains in its expression site, on a 160 kilobase (kb) chromosome, in trypanosomes not expressing the ILTat 1.3 VSG. Here we report the complete sequence of the barren region adjacent to this gene, determined both from trypanosomes expressing the gene and from those that are not. The sequence is identical whether or not the ILTat 1.3 VSG gene is expressed. This confirms that alterations in the barren region are not involved in modulation of expression of the gene, as suggested by restriction enzyme mapping. Sequence data from the 5' flanking region of a second telomeric gene copy on an 80 kb minichromosome, and from the ILTat 1.3 expression site after replacement of the ILTat 1.3 gene by another gene from a minichromosome, provide evidence that telomeric VSG genes on minichromosomes are also flanked by long repeat arrays, and that these arrays are involved in inter-telomeric gene replacements as well as replacements by non-telomeric genes. PMID- 3041211 TI - Telomeric motifs are present in a highly repetitive element in the Plasmodium berghei genome. AB - Using as probes the subfragments of the telomeric sequence previously cloned by us from Plasmodium berghei DNA, we identified and cloned a 2.3 kb repeat, largely overlapping the original telomeric insert. Restriction mapping indicated that cloned inserts (2.3 kb in length) represented circularly permutated versions of a rather well conserved repeated element, at least in part organized in tandem. The 2.3 kb repeat family with a copy number of about 300 occupies about 4% of the whole genome. The copies are unevenly distributed among the chromosome-sized molecules revealed by pulsed field gradient electrophoresis. Complete sequence determination of the 2.3 kb element revealed that telomere-related motifs are present with a characteristic pattern in a set of tandem repeats, 27 bp long. The perfect conservation of these motifs as well as the pattern of chromosomal distribution suggest that we are dealing with a specialised structure subject to selective mechanisms of amplification and maintenance. PMID- 3041212 TI - Purification and nuclear localization of a type I topoisomerase from Crithidia fasciculata. AB - A type I topoisomerase has been purified to near homogeneity from the trypanosomatid Crithidia fasciculata. The topoisomerase consists of a single 79 kDa polypeptide. The enzyme does not require divalent cations but is stimulated 10-20 fold by the presence of MgCl2. ATP does not affect enzyme activity, while Berenil, N-ethylmaleimide and ethidium bromide are inhibitory. Immunoblots show that the 79 kDa polypeptide is the most prevalent form of the enzyme in extracts of freshly lysed cells and is immunogenically conserved among a variety of trypanosomes. The topoisomerase was localized to the cell nucleus by double antibody immunofluorescence. PMID- 3041214 TI - Nitroimidazole and oxygen derived radicals detected by electron spin resonance in hydrogenosomal and cytosolic fractions from Trichomonas vaginalis. AB - Hydrogenosome-enriched fractions from Trichomonas vaginalis reduce a number of nitroimidazole derivatives to their respective electron spin resonance-detectable nitro-anion radicals. In the presence of of oxygen and 5,5-dimethyl-1-pyrroline-N oxide (DMPO) a superoxide spin trapped adduct of DMPO was formed; the rate determining step was the prior formation of the nitro-anion radical. Oxygen derived radicals were detected with cytosolic fractions from a metronidazole resistant isolate (CDC-85) when incubated with NADH or NADPH as respiratory substrate. The requirement for superoxide dismutase and catalase to completely abolish formation of these signals suggests contributions from both superoxide and peroxide. No oxygen-derived radicals were observed with cytosolic fractions from a metronidazole-susceptible strain (C1-NIH). PMID- 3041215 TI - Sequences of two kinetoplast minicircle DNAs of Trypanosoma (Nannomonas) congolense. AB - Random minicircle DNA molecules were released from isolated kinetoplast network DNA of Trypanosoma congolense by BamHI digestion and cloned into plasmid pUC19. The sequences of two cloned minicircles (958 bp and 964 bp) were determined. Both minicircles contain the 13 bp sequence, 5'-GGGGTTGGTGTAA-3', thought to be the replication origin of minicircles in other trypanosomatids. The two minicircles have extensive homology in the 120 bp preceeding, and the 20 bp following, this 13-mer but only scattered homology elsewhere. Both possess tandem repeats downstream of the 13-mer. Comparison of these minicircles with minicircle sequences from other trypanosomatids reveals that they have the same general sequence organization as the others although only the 13-mer and its flanking regions are homologous. PMID- 3041216 TI - Current concepts: immunology. Neutrophils in human diseases. PMID- 3041217 TI - High prevalence of papillomavirus-associated penile intraepithelial neoplasia in sexual partners of women with cervical intraepithelial neoplasia. AB - To determine whether neoplastic cervical lesions in women are associated with papillomavirus infections in their sexual partners, we used a colposcope to examine male sexual partners of women with cervical flat condyloma (294 cases) or cervical intraepithelial neoplasia (186 cases), before and after 5 percent acetic acid was applied to the penis and the anogenital area. Condylomata acuminata, papules, and macules were observed in 309 of the 480 men (64.4 percent). In 204 of them (42.5 percent), macules or slightly elevated papules were detected only after application of acetic acid. Condylomata acuminata or lesions showing histologic features of condyloma were found in 121 partners (41.2 percent) of women with condyloma, but in only 10 partners (5.4 percent) of women with cervical intraepithelial neoplasia. Penile lesions showing histologic features of intraepithelial neoplasia were found in 61 partners (32.8 percent) of women with cervical intraepithelial neoplasia, but in only 4 partners (1.4 percent) of women with flat condyloma. Thirty-six (60 percent) of the 60 macules or papules tested contained papillomavirus DNA sequences. Human papillomavirus types 16 and 33 were almost exclusively found in penile intraepithelial neoplasia. Type 6, type 11, and the recently recognized type 42 were found in lesions showing features of condyloma or minimal histologic changes. As yet uncharacterized papillomaviruses were found in 15 percent of the specimens. These data support the concept that cervical carcinomas and precancerous lesions in women may be associated with genital papillomavirus infection in their male sexual partners. PMID- 3041218 TI - Mutational activation of the K-ras oncogene. A possible pathogenetic factor in adenocarcinoma of the lung. AB - To define the role of cellular oncogenes in human cancers, we studied the prevalence of mutational activation of ras oncogenes in untreated non-small-cell lung cancer. Genomic DNA was extracted from 39 tumor specimens obtained by thoracotomy and was examined for activating point mutations in codons 12, 13, and 61 of the H-ras, K-ras, and N-ras genes. A novel, highly sensitive assay based on oligonucleotide hybridization following an in vitro amplification step was employed. The K-ras gene was found to be activated by point mutations in codon 12 in 5 of 10 adenocarcinomas. Two of these tumors were less than 2 cm in size and had not metastasized. No ras gene mutations were observed in 15 squamous-cell carcinomas, 10 large-cell carcinomas, 1 carcinoid, 2 metastatic adenocarcinomas from primary tumors outside the lung, and 1 small-cell carcinoma. An approximately 20-fold amplification of the unmutated K-ras gene was observed in a tumor that proved to be a solitary lung metastasis of a rectal carcinoma. We conclude that mutational K-ras activation may be an important early event in the pathogenesis of adenocarcinoma of the lung but that amplification of ras genes or mutational activation of H-ras or N-ras does not play a major part in non-small cell lung cancer. PMID- 3041219 TI - New AIDS control recommendations. PMID- 3041220 TI - Signal recognition. Two receptors act sequentially. PMID- 3041221 TI - Erythroid-specific transcription of the chicken histone H5 gene is directed by a 3' enhancer. AB - The expression of a variety of vertebrate genes is transcriptionally regulated through tissue-specific cellular enhancer elements. An erythroid-specific enhancer sequence has recently been identified 3' to (downstream of) the chicken adult beta-globin gene. Here we report the identification of a second erythroid specific enhancer sequence, which is 3' to the chicken histone H5 gene. The similarity of these two enhancer elements, with respect both to function and location relative to the target gene, implies some functional conservation in their evolution and in the mechanism by which they affect erythroid gene transcription. PMID- 3041223 TI - Neurobiology: molecules underlying memory. PMID- 3041222 TI - A signal sequence receptor in the endoplasmic reticulum membrane. AB - Protein translocation across the endoplasmic reticulum (ER) membrane is triggered at several stages by information contained in the signal sequence. Initially, the signal sequence of a nascent secretory protein upon emergence from the ribosome is recognized by a polypeptide of relative molecular mass 54,000 (Mr54K) which is part of the signal recognition particle (SRP). Binding of SRP may induce a site specific elongation arrest of translation in vitro. Attachment of the arrested translation complex to the ER membrane is mediated by the SRP-receptor (docking protein) and is accompanied by displacement of the SRP from both the ribosome and the signal sequence. We have investigated the fate of the signal sequence following the disengagement of SRP and its receptor by a crosslinking approach. We report here that the signal sequence of nascent preprolactin, after its release from the SRP, interacts with a newly discovered component, a signal sequence receptor (SSR), which is an integral, glycosylated protein of the rough ER membrane (Mr approximately 35K). PMID- 3041224 TI - Phosphotransferase sequence homology. PMID- 3041225 TI - A molecular mechanism for long-term sensitization in Aplysia. AB - Sensitization of the gill- and siphon-withdrawal reflex in Aplysia is thought to result from a set of molecular processes with different time courses: short-term sensitization is explained by cyclic AMP-dependent modulation of ion-channel function in sensory neurons lasting minutes; memory that endures for hours or longer, by the expression and distribution within the neurons of new gene products. Because gene induction and axonal transport are relatively slow, there may also be a need for a distinct form of intermediate memory to bridge the short and long-term processes. We now report that a protocol producing long-term sensitization results in a decrease in the amount of regulatory subunits of the cAMP-dependent protein kinase in animals 24 h after training, with no effect on the catalytic subunit. The loss appears to be post-translational. Because a decrease in the ratio of regulatory to catalytic subunits would result in elevated kinase activity after cAMP has returned to its unstimulated concentration in sensory cells, it could be the molecular mechanism of intermediate memory. PMID- 3041226 TI - The expression of hybrid HIV:Ty virus-like particles in yeast. AB - The yeast retrotransposon, Ty, encodes a set of proteins that are assembled into virus-like particles, Ty-VLPs (refs 1, 2). These proteins include Ty-VLP structural proteins, a protease that mediates cleavage of primary translation products and a reverse transcriptase. The major structural components of Ty-VLPs are proteolytic products of the primary translation product, p1 (ref. 3). We have recently shown that protein p1 alone can form Ty-VLPs (ref. 3). Here we demonstrate that p1 fusion proteins, comprising most of p1 and part of human immunodeficiency virus (HIV) protein gp120, form hybrid HIV:Ty-VLPs. These hybrid particles provide a rapid means of preparing and evaluating HIV antigens for a variety of immunological purposes. PMID- 3041227 TI - An intronless gene encoding a potential member of the family of receptors coupled to guanine nucleotide regulatory proteins. AB - Plasma membrane receptors for hormones, drugs, neurotransmitters and sensory stimuli are coupled to guanine nucleotide regulatory proteins. Recent cloning of the genes and/or cDNAs for several of these receptors including the visual pigment rhodopsin, the adenylate-cyclase stimulatory beta-adrenergic receptor and two subtypes of muscarinic cholinergic receptors has suggested that these are homologous proteins with several conserved structural and functional features. Whereas the rhodopsin gene consists of five exons interrupted by four introns, surprisingly the human and hamster beta-adrenergic receptor genes contain no introns in either their coding or untranslated sequences. We have cloned and sequenced a DNA fragment in the human genome which cross-hybridizes with a full length beta 2-adrenergic receptor probe at reduced stringency. Like the beta 2 adrenergic receptor this gene appears to be intronless, containing an uninterrupted long open reading frame which encodes a putative protein with all the expected structural features of a G-protein-coupled receptor. PMID- 3041228 TI - Association of DNA-bound progesterone receptors. AB - Steroid hormone-receptor complexes regulate the transcription of specific genes. Recent studies of high-affinity interactions between the receptors and discrete regions of DNA, together with gene-transfer experiments, have led to the precise mapping of hormone regulatory elements. Nothing is known, however, about the mechanisms whereby DNA-bound receptors modulate gene transcription. At the start of transcription in prokaryotes two oligomeric molecules of several regulatory proteins must bind to two specific DNA sites and interact with one another to regulate the binding of RNA polymerase to DNA. Using electron microscopy to observe progesterone receptor binding to regulatory regions of uteroglobin and mouse mammary tumour virus genes, we demonstrate a similar binding between receptor oligomers at two DNA sites. DNA loops are formed when the hormone regulatory elements are at a distance from one another. Thus, in common with certain prokaryotic systems, protein-protein interactions may be important in steroid hormone regulation of gene transcription. PMID- 3041229 TI - Site-directed mutation affecting polyomavirus capsid self-assembly in vitro. AB - Nonequivalent bonding of identical protein subunits occurs in the polyomavirus capsid were identical pentameric capsomeres occupy both hexavalent and pentavalent positions in the icosahedral surface lattice. The polyomavirus major capsid protein VP1, purified after expression of the recombinant gene in Escherichia coli, has been isolated as capsomeres that self-assemble into capsid like structures in vitro. The ability to switch bonding specificity in different symmetry environments therefore must be intrinsic to the VP1 molecule. In vitro self-assembly provides an assay for VP1 mutations affecting capsomere and capsid formation. We report here that a directed mutation in the VP1 expression vector, leading to a protein truncated at the carboxy terminus, results in a mutant VP1 that forms capsomeres, but not capsids, in the in vitro assembly assay. The carboxy terminus of VP1 therefore appears to be involved in the specific bonding responsible for the non-equivalent association of capsomeres. PMID- 3041230 TI - Antibacterial agents specifically inhibiting lipopolysaccharide synthesis. AB - The spread of antibiotic resistance in Gram-negative bacteria has sustained a continuing search for new agents with antibacterial activity against this important class of bacterial pathogen. Because the biosynthesis of lipopolysaccharide (LPS) is unique to Gram-negative bacteria and required by them for growth and virulence, attempts have been made to discover or design antibacterial agents acting at this site; however, no such agents have so far been developed. We now present definitive experimental data documenting design of the first member of the class of antibacterial compounds which specifically inhibit LPS synthesis. The target enzyme is 3-deoxy-D-manno-octulosonate cytidylytransferase (CMP-KDO synthetase), a cytoplasmic enzyme which activates 3 deoxy-D-manno-octulosonate (KDO) for incorporation into LPS. A specific inhibitor of CMP-KDO synthetase, alpha-C-(1,5-anhydro-7-amino-2,7-dideoxy-D-manno heptopyranosyl)-carboxy late was designed using results of our studies of the purified enzyme. LPS synthesis ceased and lipid A precursor accumulated, causing growth stasis and perturbation of outer membrane structure and function, following delivery of the inhibitor to the intracellular target by a peptide carrier. Antibacterial action required an intact oligopeptide permease system and specific intracellular aminopeptidase activity to release inhibitor from the peptide prodrug. PMID- 3041231 TI - Variable stoichiometry of proton pumping by the mitochondrial respiratory chain. AB - The proton/oxygen stoichiometry and the mechanism of the proton pumping respiratory complexes in the mitochondrial respiratory chain have been central issues in bioenergetics for several years. Recently, a number of disagreements about stoichiometry have been resolved, and H+/O ratios of 6 (refs 1-3) or perhaps 8 (refs 4 and 5) for succinate oxidation are now accepted. Suggestions that the stoichiometry in mitochondria is intrinsically variable ('slip' in the pumps) have been made but the evidence has been neither strong nor direct. We now show by direct measurement in steady-state conditions that the H+/O ratio (measured as the charge/O ratio) for isolated mitochondria respiring on succinate varies from 6 at low membrane potential to 2.5-3 at membrane potentials of about 170 mV. PMID- 3041232 TI - Discrimination between antibodies to HIV and to related retroviruses using site directed serology. AB - Human immunodeficiency virus (HIV) strains can be separated into two types: HIV and HIV-related West African viruses. Site-directed serology using synthetic peptides offers possibilities for the determination of type-specific antibodies. A 22-amino-acid peptide with the sequence Ala-Ile-Glu-Lys-Tyr-Leu-Glu-Asp-Gln-Ala Gln-Leu-Asn-Ala-Trp-Cys-Ala-Phe-Arg-Gln - Val-Cys representing a conserved region of the transmembranous protein of simian T-cell lymphotropic virus-type III (STLV III; related to West African HIV) was used as antigen in an enzyme-linked immunosorbent assay (ELISA). In parallel, tests were performed with a pair of previously described peptides, including the homologous region of the glycoprotein (gp) 41 of the HIV strain HTLV-IIIB. In tests with three groups of 20 sera it was shown that the different peptide ELISAs allowed a categorical distinction of antibodies to the two types of HIV. Tests using peptide antigens may provide excellent opportunities for large-scale testing for type-specific antibodies against HIV. The tests are simple, sensitive and specific and are readily standardized. PMID- 3041234 TI - Enzyme identity maintained. PMID- 3041233 TI - Functionally distinct subsites on a class II major histocompatibility complex molecule. AB - Mature T lymphocytes are activated by recognition of the combination of foreign protein antigen and membrane products of the major histocompatibility complex (MHC). Studies of peptide antigen binding to detergent-solubilized class II MHC molecules (Ia) have established that peptide-Ia interaction occurs in the absence of the T-cell receptor and varies according to allele-specific features of Ia molecules. The residues of immunogenic peptides thus contribute to two largely independent functions--the control of association with Ia molecules and the determination of the specificity of T-cell receptor binding. Two analogous and potentially independent functional sites have been postulated for Ia molecules--a region that controls binding to peptides and a region that interacts with T-cell receptors. Here we present evidence from functional analysis of recombinant class II molecules that these two postulated functional regions of Ia molecules do exist and can be independently manipulated, consistent with our recent demonstration of the segmental nature of Ia molecule structure-function relationships. PMID- 3041235 TI - First Conference on DNA Topoisomerases in Cancer Chemotherapy. New York, N.Y., November 19-20, 1986. PMID- 3041236 TI - DNA topoisomerase II as a potential factor in drug resistance of human malignancies. AB - The stabilization of the cleavable complex between DNA topoisomerase II and DNA by adriamycin (ADR), as well as by other topoisomerase II-targeted drugs, is an essential step in a process associated with drug cytotoxicity. Unlike many other cell types, ADR does not produce DNA cleavage in the lymphocytes of chronic lymphocytic leukemia (CLL). The CLL lymphocytes have been identified as quiescent cells with an extremely low level of topoisomerase II. The low level of this enzyme could constitute a basis for a new mechanism of drug resistance operating not only in CLL, but perhaps in any slow growing cancer with a large population of quiescent cells. Other factors contributing to drug resistance could include changes in enzyme regulation or processing of the cleavable complex, or the presence of a "mutant" enzyme which renders cancer cells unresponsive to topoisomerase II-targeted drugs. Suggested strategies in drug development, aimed at the topoisomerase II-related drug resistance, could include 1) the selection of topoisomerase I as an alternative target for cancer chemotherapy, 2) the development of ADR analogs which, unlike ADR, stabilize the topoisomerase II-DNA complex with high efficiency, and 3) the search for agents enhancing the SOS-like repair response, presumably triggered by DNA topoisomerase-targeted drugs. PMID- 3041237 TI - Metabolic activation of N-acylanthracyclines precedes their interaction with DNA topoisomerase II. AB - The N-acylanthracyclines AD32 (N-trifluoroacetyladriamycin-14-valerate) and AD143 (N-trifluoroacetyladriamycin-14-O-hemiadipate) are analogs of Adriamycin (ADR) undergoing clinical or advanced pre-clinical screening. Their principal metabolites, following the cleavage of the 14-acyl side-chain, are N trifluoroacetyladriamycin (AD41) and its reduced form N trifluoroacetyladriamycinol (AD92). Both these compounds are biologically active and detectable in treated patients, laboratory animals, and in tissue culture cells. Unlike ADR, AD32, as well as AD143 and metabolites, show no detectable binding to double-strand DNA. Their effects on DNA have been previously investigated in vivo and in vitro using the alkaline filter-elution assay. It has been shown that all of the compounds cause approximately equivalent amounts of protein-associated DNA breaks (PAB) and DNA-protein crosslinks in a mouse lymphoma and in tissue-culture leukemia cells. In order to establish whether the induction of PAB by the drugs requires DNA topoisomerase II mediation, cleavage mapping analysis was done with tested compounds using purified human topoisomerase II. DNA fragmentation was significantly enhanced in the presence of the enzyme and either AD41 or AD92. In contrast, no fragmentation enhancement was observed in the presence of the parental drugs AD32 or AD143. The results strongly suggest that metabolic activation of N-acylanthracyclines by nonspecific esterases is a prerequisite for their interaction with DNA topoisomerase II and for stabilization of the cleavable complex. PMID- 3041238 TI - Protein-linked DNA strand breaks produced by etoposide and teniposide in mouse L1210 and human VA-13 and HT-29 cell lines: relationship to cytotoxicity. AB - The two demethylepipodophyllotoxin glycosides, teniposide (VM-26) and etoposide (VP-16), have previously been reported to interact with DNA topoisomerase II by stabilizing a topoisomerase II-DNA covalent intermediate. This study examined the protein-associated aspect of the topoisomerase II-DNA-epipodophyllotoxin lesion. We found that in mouse (L1210) and human (VA-13 and HT-29) log-phase cell cultures, all DNA strand breaks produced by VP-16 or VM-26 were protein associated. We found also that these protein-associated breaks occurred with a frequency which correlated with cytotoxicity in all three cell lines. For all three cell lines and for both compounds the regression lines were similar. Therefore, for a given class of topoisomerase II inhibitors, it may be possible to generate a characteristic line from which DNA-protein crosslink frequency predicts cytotoxicity. PMID- 3041239 TI - Structure-activity relationships of podophyllin congeners that inhibit topoisomerase II. AB - Various analogs of etoposide have been studied and compared in different tests in order to identify which tests best correlate with antitumor activity. These tests included DNA breakage assays using standard alkaline elution procedures as a means of studying topoisomerase II inhibition in intact cells, cytotoxicity studies in naturally sensitive and resistant human carcinoma cell lines, in vitro assays of the effect of the different congeners on topoisomerase II activity, and a preliminary evaluation of the ability of etoposide and teniposide to induce resistance. As in previous studies, a direct correlation was seen between double strand DNA breakage and cytotoxicity but not between single strand DNA breakage and cytotoxicity. Analogs with blocked 4'-hydroxyl groups were poor antitumor agents but were still capable of inhibiting topoisomerase II as evidenced by the production of DNA breaks. However, this DNA breakage was qualitatively different from that produced by VP16. None of the analogs were able to overcome either naive or acquired drug resistance. The dihydroxy analog of VP16, a possible bioactivated analog, was much less potent and possibly less stable than VP16. A model is proposed for the inhibition of topoisomerase II by demethylepipodophyllotoxins that may explain the relationship between double strand DNA breakage and cytotoxicity. PMID- 3041240 TI - Transcription of a Rhizobium leguminosarum biovar phaseoli gene needed for melanin synthesis is activated by nifA of Rhizobium and Klebsiella pneumoniae. AB - The Rhizobium leguminosarum biovar phaseoli symbiotic plasmid pRP2JI carries a gene, melA, specifying the enzyme tyrosinase, which is responsible for the production of the pigment melanin in these bacteria. Transcription of melA is activated by the nifA gene of Rhizobium and, when the cloned melA gene is transferred to Escherichia coli, melA is expressed if the recipients contain nifA gene of Klebsiella pneumoniae. This nifA-dependent activation was temperature sensitive and required the ntrA gene. The cloned nifA gene of K. pneumoniae, when transferred to a nifA mutant of Rhizobium phaseoli biovar phaseoli, corrected the Mel- but not the Fix- phenotype. nifA of R. leguminosarum biovar phaseoli activated melA at higher levels in cells grown in low concentrations of oxygen. Also, nifA of R. leguminosarum biovar phaseoli activated nifH of K. pneumoniae in Escherichia coli cells grown in low-oxygen concentrations. PMID- 3041242 TI - Repression of the aroF promoter by the TyrR repressor in Escherichia coli K-12: role of the 'upstream' operator site. AB - Three sequences are required for complete repression of the aroF promoter by the TyrR repressor protein. Two of these operator sites lie adjacent to each other and overlap the -35 region of the aroF promoter while the third lies about 70 base pairs upstream of the promoter. An aroF-cat (chloramphenicol acetyltransferase) gene fusion has been used to assay the effect of DNA insertions that alter the distance between the two promoter-proximal and the third, distal, operator sites on the repression of the aroF promoter in vivo. The distal site contributes to the repression of the promoter up to a distance of about 400 base pairs and its effect is not dependent on its separation from the first and second sites by an integral number of turns of the DNA helix. PMID- 3041241 TI - Catabolite control of the elevation of PGK mRNA levels by heat shock in Saccharomyces cerevisiae. AB - Heat shock enhances the very high level of transcription of the phosphoglycerate kinase (PGK) gene in fermentative cultures of Saccharomyces cerevisiae. This response of PGK mRNA levels was not found on gluconeogenic carbon sources, and could be switched on or off subject to availability of fermentable carbon source. The addition of glucose to yeast growing on glycerol resulted in acquisition, within 30-60 min, of the ability to elevate PGK mRNA levels after heat shock. In addition, in aerobic cultures growing on glucose the exhaustion of the medium glucose coincided with a loss of the heat-shock effect on PGK mRNA and a switch over to slower growth by aerobic respiration. Levels of hsp26 mRNA were analysed during these experiments. Contrasting with this requirement for fermentable catabolite for manifestation of a heat-shock response of PGK mRNA levels, the PGK enzyme was not synthesized at a greater level in heat-shocked fermentative than in gluconeogenic cultures. PGK is one of only a few proteins made efficiently after mild heat shock of yeast. Thus, heat-stress-induced elevation of PGK mRNA levels does not appreciably increase PGK synthesis during exposure to high temperatures and so its role may be to assist cells repressed in mitochondrial function during recovery following a heat shock. PMID- 3041243 TI - Covalent modification of ribulose 1,5-bisphosphate carboxylase/oxygenase in Rhodomicrobium vannielii. AB - The most abundant phosphorus-containing polypeptide in the purple non-sulphur bacterium Rhodomic-robium vannielii has been identified by a combination of immunoprecipitation and sucrose density gradient centrifugation as the large subunit of ribulose 1,5-bisphosphate carboxylase/oxygenase. The covalent modification of the large subunit involves the phosphorylation of one or more tyrosine residues and appears to occur prior to assembly of the large subunit into the mature enzyme. In addition, the phosphorylated form of the large subunit was found to exist in at least two distinct protein complexes of Mr 410,000 and 440,000. PMID- 3041244 TI - Acute ethanol poisoning and the ethanol withdrawal syndrome. AB - Ethanol, a highly lipid-soluble compound, appears to exert its effects through interactions with the cell membrane. Cell membrane alterations indirectly affect the functioning of membrane-associated proteins, which function as channels, carriers, enzymes and receptors. For example, studies suggest that ethanol exerts an effect upon the gamma-aminobutyric acid (GABA)-benzodiazepine-chloride ionophore receptor complex, thereby accounting for the biochemical and clinical similarities between ethanol, benzodiazepines and barbiturates. The patient with acute ethanol poisoning may present with symptoms ranging from slurred speech, ataxia and incoordination to coma, potentially resulting in respiratory depression and death. At blood alcohol concentrations of greater than 250 mg% (250 mg% = 250 mg/dl = 2.5 g/L = 0.250%), the patient is usually at risk of coma. Children and alcohol-naive adults may experience severe toxicity at blood alcohol concentrations less than 100 mg%, whereas alcoholics may demonstrate significant impairment only at concentrations greater than 300 mg%. Upon presentation of a patient suspected of acute ethanol poisoning, cardiovascular and respiratory stabilisation should be assured. Thiamine (vitamin B1) and then dextrose should be administered, and the blood alcohol concentration measured. Subsequent to stabilisation, alternative aetiologies for the signs and symptoms observed should be considered. There are presently no agents available for clinical use that will reverse the acute effects of ethanol. Treatment consists of supportive care and close observation until the blood alcohol concentration decreases to a non-toxic level. In the non-dependent adult, ethanol is metabolised at the rate of approximately 15 mg%/hour. Haemodialysis may be considered in cases of a severely ill child or comatose adult. Follow-up may include referral for counselling for alcohol abuse, suicide attempts, or parental neglect (in children). The ethanol withdrawal syndrome may be observed in the ethanol-dependent patient within 8 hours of the last drink, with blood alcohol concentrations in excess of 200 mg%. Symptoms consist of tremor, nausea and vomiting, increased blood pressure and heart rate, paroxysmal sweats, depression, and anxiety. Alterations in the GABA benzodiazepine-chloride receptor complex, noradrenergic overactivity, and hypothalamic-pituitary-adrenal axis stimulation are suggested explanations for withdrawal symptomatology.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3041245 TI - Hepatotoxicity of methotrexate in rheumatic diseases. AB - Methotrexate-induced hepatotoxicity is well recognised in the treatment of leukaemia, psoriasis and rheumatoid arthritis. The pathological lesions are non specific, consisting of fatty change, nuclear pleomorphism, hepatocyte necrosis, portal chronic inflammatory infiltrate, fibrosis and cirrhosis. The mechanism of liver injury is poorly understood; intracellular accumulation of methotrexate polyglutamate and consequent folate depletion are suspected to play a role. Early studies in psoriasis clearly established a relationship of the hepatic injury with the frequency of methotrexate administration. With weekly low dose therapy, however, consensus is lacking regarding the incidence of hepatotoxicity because studies have had disparate control groups, used variable dosage regimens and often failed to document pre-existing liver disease or categorised patients at risk, i.e. elderly patients, alcoholics and obese diabetics. Moreover, current methods of assessing the degree of hepatic injury are subjective, relying on interpretation by an experienced histopathologist. Preliminary evidence suggests less frequent and less severe hepatotoxicity occurs in patients with rheumatoid arthritis, probably as a result of lower methotrexate doses and better patient selection. Nevertheless, until the risk of serious liver disease is better defined it is recommended that patients have a pretreatment liver biopsy, a follow-up biopsy after a cumulative dose of 1500 mg, and then biopsies approximately every 2 years in the absence of other evidence of liver disease or risk factors. PMID- 3041247 TI - Menorrhagia and menopause: a historical review. AB - Excessive premenopausal uterine bleeding, whether an exaggeration of catamenial loss or more severe haemorrhage, has until lately been part of traditional climacteric symptomatology, yet it is no longer so. This, and a parallel article, attempt to find out the reasons for the lapse of this symptom. The present paper concentrates on the literature of the 18-19th century, generously quoting selected sources, in an effort to define the context in which this symptom became so prominent, the explanations offered by and the approach of contemporaries to it. The Classical ideas concerning the menses and their cessation, briefly outlined, were challenged during the 18th century by a few individuals who attempted to ascertain the facts for themselves and establish what was the actual course of nature. They were helped in this by the presence of differing cultural subgroups with completely different climacteric experiences. The conviction then arose that medications given upper class women were iatrogenically responsible for the complications they experienced. When menorrhagia continued the lifestyle of these ladies was blamed. Despite all corrections, however, menorrhagia persisted. This review then examines, with varying detail, some of the writings of these two centuries, offering some glimpses into a literature otherwise not easily accessible. PMID- 3041246 TI - Pharmaceutical excipients. Adverse effects associated with 'inactive' ingredients in drug products (Part II). PMID- 3041250 TI - Psychosocial aspects of cardiac transplantation. PMID- 3041249 TI - Bronchodilator therapy in cystic fibrosis: for better or worse? AB - As one approach to the management of the respiratory manifestations of cystic fibrosis, bronchodilators have been used to try to alleviate symptomatology. Several bronchodilators have been tested with varying degrees of success depending on the parameters measured, the method of patient selection for trials and the time of the study. In some instances the use of bronchodilators has proven detrimental to lung function. In general, it appears that bronchodilator therapy may be beneficial or hazardous and must be evaluated on an individual patient basis. Repeated assessments of respiratory function are necessary and the patient's management must be frequently re-evaluated. PMID- 3041248 TI - Continuous oestrogen-progestogen treatment and bone metabolism in post-menopausal women. AB - The suggestion that norethisterone acetate (NETA) stimulates bone formation prompted us to investigate calcium and bone metabolism in post-menopausal women treated with a continuous combination of oestrogen and NETA. The study included 49 healthy post-menopausal women. After an initial examination the women were randomly allocated to 2 yr of treatment with either hormones or placebo; 43 women completed the study. Within the first year of treatment the hormone group showed a slight, but significant, increase in bone mineral content in the forearms (single photon absorptiometry) and in the total skeleton (dual photon absorptiometry). After this initial increase the bone mass remained constant throughout the trial. Bone mineral density of the spine (dual photon absorptiometry) showed a significant increase of more than 5% in the hormone group. There were significant decreases of 4-7% in bone mass in the placebo group over 2 yr. Biochemical estimates of bone turnover (plasma bone Gla protein, serum alkaline phosphatase, fasting urinary calcium and fasting urinary hydroxyproline) fell significantly in the hormone group, but remained unchanged in the placebo group. We conclude that continuous administration of NETA in combination with oestrogen provides effective prophylaxis against post-menopausal bone loss, although it does not produce a persistent positive calcium balance in early post menopausal women. PMID- 3041251 TI - Psychological adjustment after cardiac transplantation. AB - Cardiac transplantation is viable therapeutic alternative for patients with end stage heart disease, which offers a favourable short- and medium-term prognosis. The survival has improved from 20% of patients who survived at one year after transplantation in the 1960s to the present figures of 80%-85% of patients who are alive at one year, and 50%-70% of patients who are alive at five years, after transplantation. Therefore, it seems timely to focus attention on the psychological well-being of cardiac-transplant recipients. The medical literature is scant in regard to the psychiatric and the psychosocial impact of cardiac transplantation on recipients, and a systematic and prospective study of the psychosocial adaptation of recipients is lacking. Since 1984, we have been studying the emotional impact of cardiac transplantation on recipients and their families. This article presents the results for a group of recipients who have been assessed before transplantation, then followed-up at discharge from hospital and at four, eight and 12 months after transplantation. The study attempted to quantitate the recipients' anxiety, depression, body image and subjective quality of life by way of standardized self-assessment questionnaires. The recipients' satisfaction with relationships or their marital situation also was reported, as were their degree of rehabilitation at 12 months and their attitudes to various aspects of treatment after the transplantation. Before the transplantation, 53% of patients reported an increase in anxiety and 34% of patients recorded scores that indicated mild-to-moderate levels of depression. Thirty-seven per cent of patients showed a deterioration in the quality of their lives and 34% of patients had a negative body image. After the transplantation, significant improvements occurred in all parameters, which were maintained at follow-up. PMID- 3041252 TI - Oxygen therapy. PMID- 3041253 TI - [The antimycotic ketoconazole. Indications in prostate disease, hirsutism, precocious puberty and Cushing syndrome?]. PMID- 3041254 TI - [Before or after eating? Drug interactions with food]. PMID- 3041255 TI - [Stomach and duodenal ulcer. Pathogenesis and conservative therapy]. PMID- 3041256 TI - [Cytostatic therapy of melanoma: systemic or regional? Pharmacologic principles, results up to now and current developments]. PMID- 3041257 TI - [Physiology and pathophysiology of stomach emptying. Principles, methods of study and therapy]. PMID- 3041258 TI - [Reflections on the place of polygraphic recordings in the study of hypnotic drugs]. AB - Polygraphic recordings are important for studying a new hypnotic molecule. Night recordings are used in the assessment of the objective hypnotic effect over short and long-term periods. They show sleep architecture modifications caused by the hypnotic drug and quantify rebound insomnia following abrupt withdrawal. The comparative usefulness of sleep laboratory recordings and ambulatory monitoring is discussed. Polygraphic recordings also allow to evaluate diurnal residual effect upon vigilance. Twenty-four-hour recordings as well as multiple sleep latency test (MSLT) are used. PMID- 3041259 TI - The role of water in cell architecture. AB - The role of water in biochemical and cellular events is ignored by most workers. However, much recent research has pointed to the importance of physical processes of the cell, which focus attention on such straight forward, elementary questions as position and relationship in space of cell components. In this communication these questions are examined in terms of a new model of water structure. A radically new feature of this model is that water clusters have long-term rather than flickering existence and are as large as the macromolecular components of the cell. These properties allow the clusters and other components to pack together spacially so giving rise to integrated, large-scale, subcellular structures. The intimate participation of water in these structures would explain the fragility of the cytoplasmic organization. PMID- 3041260 TI - Processing of the luteinizing hormone-releasing hormone precursor in the preoptic area and hypothalamus of the rat. AB - The LHRH precursor is known to contain the decapeptide and a 56 amino acid peptide termed gonadotropin-releasing hormone-associated peptide (GAP). The purpose of our study was to characterize the proLHRH and its processed products from the cell body and fiber region and from the nerve terminal region of LHRH neurons. The median eminence (ME) and a tissue block containing the preoptic area and hypothalamus (POH) were dissected separately. Tissues were homogenized and peptides were separated according to mol wt. Three different LHRH antisera bound to one immunoreactive (IR) substance which eluted at approximately 1200 mol wt. Subsequently, this material coeluted with synthetic LHRH on a reversed-phase column as a single peak. There was approximately 1.6-fold more LHRH-like IR in the ME than in the POH. The four different GAP antisera recognized multiple mol wt forms of GAP-like IR at approximately 16,000 to 14,000, 8,200, 6,500, 3,500, and 2,800 mol wt. There were more of the high mol wt materials and less of the 6500 and lower mol wt materials in the POH than in the ME. The most abundant species in both regions was the 6500 mol wt form. This IR substance coeluted with synthetic rat GAP1-56 on a reversed-phase column as a single peak. These experiments demonstrate 1) that multiple IR forms of the LHRH prohormone exist in the POH of the rat and 2) that nerve terminals of the LHRH neurons contain LHRH, GAP1-56, and some lower mol wt GAP-like substances. These results provide the first information concerning the processing scheme for the LHRH prohormone in the rat brain. PMID- 3041261 TI - In memoriam J.S.L. Browne, M.D., Ph.D. PMID- 3041262 TI - Multihormonal regulation of insulin-like growth factor-I-related protein in MCF-7 human breast cancer cells. AB - MCF-7 human breast cancer cells have been studied for hormonal regulation of secretion of an insulin growth factor-I (IGF-I)-related growth factor. 17 beta Estradiol, which is required for tumorigenesis of the cell line in the nude mouse and which stimulates proliferation in vitro, was able to significantly induce IGF I secretion at 10(-13) M, with maximal induction at 10(-11) M. Under optimal conditions IGF-I could be induced 4-fold after 4 days. Demonstration of estrogenic stimulations required removal of phenol red, a weak estrogen, from the cell culture medium. In addition to estrogen, insulin, epidermal growth factor, and transforming growth factor alpha induce both cellular proliferation and IGF-I secretion, while growth inhibitory antiestrogens, transforming growth factor beta, and glucocorticoids have the opposite effect. In each case, modulations in IGF-I secretion preceeded effects on cellular proliferation. IGF-I was not regulated by human GH, basic fibroblast growth factor, platelet-derived growth factor, or PRL, none of which affected proliferation rate. Thus, regulation of IGF-I secretion in human breast cancer is controlled by different hormones from those previously reported in human fibroblasts. Regulation of IGF-I by neither estrogen nor antiestrogen was associated with changes in steady-state mRNA levels; thus regulation may occur at a step beyond mRNA. We conclude that IGF-I production is tightly coupled to growth regulation by estrogens, antiestrogens, and other hormones and may contribute to autocrine and/or paracrine growth regulation by these agents in breast cancer. PMID- 3041264 TI - Current concepts in treating elderly patients: a decade of advances. PMID- 3041263 TI - In memoriam: Edward H. Rynearson, M.D. PMID- 3041265 TI - Heart failure in the elderly. PMID- 3041266 TI - Anxiety and the cardiovascular system. PMID- 3041267 TI - Special rheumatologic problems of older people. PMID- 3041268 TI - Avoiding adverse drug reactions in the elderly. PMID- 3041269 TI - Surveillance for colonic polyps. PMID- 3041270 TI - Antimutagenic effect of volatile decomposition products from thermally oxidized linoleate. AB - The antimutagenic effect of volatile decomposition products from thermally oxidized linoleate on mutagenesis by UV irradiation was investigated in Escherichia coli B/r WP2. When added to an agar medium, these products greatly reduced the number of Trp+ revertants. The same antimutagenic effect was observed by acrolein, 2-hexenal, 2-heptenal, 2-nonenal and 2,4-decadienal; these unsaturated aldehydes were components of volatile products. PMID- 3041271 TI - Overproduction of single-stranded DNA-binding protein increases UV-induced mutagenesis in Escherichia coli. AB - UV-induced mutagenesis was investigated in the uvrB strain and its isogenic counterpart overproducing the single-stranded DNA-binding protein (SSB). It was demonstrated that overproduction of SSB significantly increases the frequency of mutation. Our results indicate that such an increase might be due to certain abnormalities in induction of the SOS response (untimely and prolonged activation of the RecA protein). PMID- 3041272 TI - Mutagenicity testing of seminal fluid: seminal fluid increases the mutagenicity of the precursor mutagen benzo[a]pyrene in the presence of S9 mix. AB - Small amounts of seminal fluid strongly enhanced the mutagenicity of the precursor mutagen benzo[a]pyrene (BP) in the Salmonella/microsome test. This previously unreported effect was found only in the presence of S9 mix for metabolic activation. The increase far exceeded the additive effect expected from experiments where seminal fluid and BP were tested separately with S9 mix. Testing of the direct-acting mutagen 4-nitro-o-phenylene-diamine (NPD) together with seminal fluid resulted in a lower mutagenic activity than that of NPD alone. Seminal fluid had a bactericidal effect on the Salmonella bacteria, thus only volumes up to 40 microliter could be used per plate. The mutagenic effect of only seminal fluid and S9 mix was slightly increased over controls in a standard Ames test, but was equal to the spontaneous mutation rate with a preincubation test modified according to Kado and coworkers. There were no significant differences between seminal plasma from smokers and non-smokers in any experimental series. Seminal fluid concentrated 20-fold by extraction with the mutagen-removing adsorbant Mutasorb did not have any enhancing effect on the mutagenicity of BP, nor did it exhibit any mutagenic activity in itself with or without S9 mix. PMID- 3041273 TI - Butylated hydroxyanisole, butylated hydroxytoluene and tert.-butylhydroquinone are not mutagenic in the Salmonella/microsome assay using new tester strains. AB - The phenolic antioxidants butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT) and tert.-butylhydroquinone (TBHQ) were reassessed for mutagenic activity using the recently developed Salmonella tester strains TA97, TA102 and TA104, and in addition TA100. None of the phenolic antioxidants showed mutagenic activity, either with or without metabolic activation. At doses of 100 micrograms/plate and higher all 3 phenolic antioxidants exhibited toxic effects. A modification of the assay using the preincubation procedure with strain TA104 did not affect mutation frequencies. Combinations of BHA and BHT, tested to detect possible synergistic effects, did not exert mutagenic activity. PMID- 3041274 TI - The mutagenic and clastogenic activity of tobacco smoke. AB - Employing the Salmonella/microsome mutagenicity assay it was established that the mutagenic effect of tobacco smoke (TS) (240 cm3 in a 16-l glass chamber, at 1 min or 5 min exposure time) in S. typhimurium TA98 depended on the type of S9 mix used. Addition of S9 mix obtained from the liver of 3-methylcholanthrene- or Aroclor-1254-pretreated rats but not from the liver of phenobarbital-pretreated or untreated rats was required to demonstrate the mutagenic activity of TS. One might suggest that polycyclic aromatic hydrocarbons were involved in TS-induced mutagenesis in S. typhimurium TA98. In addition, treatment of BDF1 mice with TS (600 cm3 TS in a 14-l glass chamber, 2-6 exposures of 30 min each with a 1-min interval between them during which a total change of the air was made) caused an up to 3.5-fold increase of the number of micronucleated polychromatic erythrocytes (PCE) in mouse bone marrow detected 24 h after the TS exposure. Furthermore, a stable 2-5-fold elevation of the number of micronucleated normochromatic erythrocytes (NCE) was detected in the peripheral blood of mice treated daily (2 x 30 min) with TS, starting 48 h after the first TS exposure. The application of the micronucleus test in mouse peripheral blood, a more convenient and useful approach for detecting the chronic clastogenic activity of TS, allowed us to establish the cumulative genotoxic effect of TS in mice. PMID- 3041275 TI - Schwann cell-conditioned medium supports neurite outgrowth and survival of spinal cord neurons in culture. AB - The effect of Schwann cell-conditioned medium (SCM) on the development in vitro of spinal cord neurons was studied. Spinal cord neurons from 18-day-old rat embryos were cultured in serum-free conditioned medium obtained from confluent rat Schwann cells. In cultures fed SCM, the cells developed typical neuronal morphology and were identified by indirect immunofluorescence using a monoclonal antibody to neurofilament protein. SCM stimulated neurite outgrowth and supported survival of spinal cord neurons. Preliminary characterization suggests that the neurotrophic factor in SCM appears to be a protein with a molecular weight greater than 8000 daltons. PMID- 3041276 TI - Purification and characterization of proteases secreted by transforming schistosomula of Schistosoma mansoni. AB - Schistosomula of Schistosoma mansoni which are mechanically transformed at 4 degrees C and are then incubated at 37 degrees C in defined medium spontaneously secrete two proteases, a major one of 28 kDa and a minor one of 60 kDa. These were purified by ion exchange chromatography on DEAE-cellulose and gel filtration on Ultrogel AcA 54 with yields of 33% and 29%, respectively. Both appeared as single bands by silver staining following sodium dodecyl sulphate-polyacrylamide gel electrophoresis analysis. The 28 kDa protease is a glycoprotein that has a pI of 11 or higher and an optimal activity around pH 9.0. It cleaves casein, gelatin and human C3 and C3b. It is metal-ion independent and is inhibited by diisopropyl fluorophosphate, phenylmethanesulfonyl fluoride, soy-bean trypsin inhibitor, alpha 1 antitrypsin, Zn2+ ions, sodium dodecyl sulphate and normal human serum. The 60 kDa protease is a glycoprotein with a pI of 9.2. It can also cleave casein and gelatin and its activity is inhibited by phenylmethanesulfonyl fluoride but not by diisopropyl fluorophosphate or sodium dodecyl sulphate. We suggest that these proteases may play a role during cercarial penetration of the skin and in shedding of the cercarial glycocalyx. PMID- 3041277 TI - Cryptic disposition of antigenic parasite proteins in plasma membranes of erythrocytes infected with Plasmodium chabaudi. AB - Plasma membranes of Plasmodium chabaudi-infected erythrocytes contain seven major neoproteins with apparent molecular masses of 154, 145, 90, 72, 67, 52, and 33 kDa, respectively. These neoproteins, with the exception of the two larger ones, can be metabolically labelled with [14C]isoleucine. The seven neoproteins are antigenic as revealed by Western blotting using hyperimmune sera obtained from two different mouse strains. None of the parasite proteins is accessible from the outside in intact P. chabaudi-infected erythrocytes as determined by lactoperoxidase-mediated radioiodination, indirect immune fluorescence microscopy, or post-embedding immunoelectron microscopy. These methods, however, identify parasite proteins in host cell plasma membranes when the latter are artificially changed either during isolation or by methanol fixation. We conclude therefore that parasitic proteins are cryptically arranged in intact host cell plasma membranes of P. chaubaudi-infected erythrocytes. PMID- 3041278 TI - Oxidant defense enzymes of Plasmodium falciparum. AB - We have measured and characterized three oxidant defense enzymes in early and late intraerythrocytic stages of the human malarial parasite, Plasmodium falciparum. Isolated early intraerythrocytic stages contain catalase (24.1 mumol min-1 (mg protein)-1) and superoxide dismutase (SOD; 6.3 units (mg protein)-1) but little or no glutathione peroxidase (GPX; less than 2 mumol min-1 (mg protein)-1). Isolated late intraerythrocytic stages of P. falciparum contain slightly less catalase (17.0 mumol min-1 (mg protein)-1) but significantly more GPX (7.7 mumol min-1 (mg protein)-1) and SOD (25.1 units (mg protein)-1). P. falciparum, like P. berghei, probably acquires most of its SOD from its host, since parasite-associated SOD is predominantly cyanide-sensitive, and has the same pI as host SOD. Unlike P. berghei, however, late stages of P. falciparum contain an additional SOD isozyme which is not cyanide-sensitive and may represent an endogenous enzyme. Parasites grown in red cells that have been partially depleted of SOD are more sensitive to exogenously generated superoxide, suggesting some dependence of the parasite on host SOD. PMID- 3041279 TI - Clinical and biochemical manifestations of depression. Relation to the neurobiology of stress (2) AB - Thousands of studies have been conducted of the functioning of the many neurotransmitter systems in order to explore the biologic basis of major depressive disorder. Instead of reviewing this literature exhaustively, we have attempted to propose a model that accommodates the clinical observation that chronic stress early in life in vulnerable persons predisposes them to major depression with contemporary observations of the potential consequences of repeated central nervous system exposure to effectors of the stress response. This model accords with current clinical judgment that major depression is best treated with a combination of psychopharmacologic agents and psychotherapy. Accordingly, whereas psychopharmacologic intervention may be required to resolve an active episode of major depression and to prevent recurrences, psychotherapy may be equally important to lessen the burden of stress imposed by intense inner conflict and counterproductive defenses. PMID- 3041280 TI - Hoechst wins battle. PMID- 3041281 TI - New genome money. PMID- 3041283 TI - Deactivation of macrophages by transforming growth factor-beta. AB - Macrophage activation--enhanced capacity to kill, in a cell that otherwise mostly scavenges--is essential for host survival from infection and contributes to containment of tumours. Both microbes and tumour cells, therefore, may be under pressure to inhibit or reverse the activation of macrophages. This reasoning led to the demonstration of macrophage deactivating factors from both microbes and tumour cells. In some circumstances the host itself probably requires the ability to deactivate macrophages. Macrophages are essential to the healing of wounds and repair of tissues damaged by inflammation. Yet the cytotoxic products of the activated macrophages can damage endothelium, fibroblasts, smooth muscle and parenchymal cells (reviewed in ref. 6). Thus, after an inflammatory site has been sterilized, the impact of macrophage activation on the host might shift from benefit to detriment. These concepts led us to search for macrophage deactivating effects among polypeptide growth factors that regulate angiogenesis, fibrogenesis and other aspects of tissue repair. Among 11 such factors, two proteins that are 71% similar proved to be potent macrophage deactivators: these are transforming growth factor-beta 1 (TGF-beta 1) and TGF-beta 2. PMID- 3041282 TI - Spliceosomal RNA U6 is remarkably conserved from yeast to mammals. AB - The small nuclear RNA U6 and its gene have been isolated from yeast. In striking contrast to other yeast spliceosomal RNAs, U6 is very similar in size, sequence and structure to its mammalian homologue. The single-copy gene is essential. These properties suggest a central role in pre-mRNA processing. An extensive base pairing interaction with U4 snRNA is described; the destabilization of the U4/U6 complex seen during splicing thus requires a large conformational change. PMID- 3041284 TI - [Chimeras in biologic embryology]. AB - Chimeras produced from amphibian, mammalian, and especially avian embryos have provided important insights into vertebrate development. Important contributions have led to new concepts in understanding the development of, for example, the nervous system, the vascular system, and the skeletal muscles. The migration of cells is particularly accessible in chimeras. More important results are to be expected from chimeras in the future, especially by combining this approach with other state-of-the-art techniques. PMID- 3041285 TI - Liver transplantation. Initiation of the first program in North Carolina. PMID- 3041287 TI - Richard Gatling and his gun. PMID- 3041286 TI - "Stuart factor" (coagulation factor X). A North Carolina saga. PMID- 3041288 TI - [Corticosteroids]. PMID- 3041289 TI - [Forms of pericarditis; symptoms, diagnosis and therapy]. PMID- 3041290 TI - [Experiences with amebic liver abscess at the Harbor Hospital 1978-1986]. PMID- 3041291 TI - [Lung embolism from a diagnostic perspective]. PMID- 3041293 TI - Adult dominant polycystic kidney disease--clinical problems. PMID- 3041292 TI - [Sources of thalamocortical afferentation of the area of forelimb representation in the first somatosensory zone of the cerebral cortex in the cat]. AB - Retrogradely labelled thalamic neurons--the sources of afferentation in the focus of maximal activity evoked by stimulation of n. radialis--were studied in adult cats by means of microiontophoretic injections of horseradish peroxidase into the first somatosensory zone of the cerebral cortex. The labelled cells were predominantly found in the thalamic nucleus ventralis posterolateralis; some cells (a minor part) were also found in the nucleus ventralis posteromedialis. The labelled neurons in these nuclei were distributed in groups whose composition according to many morphological characteristics was heterogeneous. PMID- 3041294 TI - Biochemical and serological characteristics of children with membranous nephropathy due to hepatitis B virus infection: correlation with hepatitis B e antigen, hepatitis B DNA and hepatitis D. AB - Fourteen children with biopsy-proven membranous nephropathy associated with hepatitis B virus (HBV-MN) were evaluated biochemically and serologically and compared to 45 children with idiopathic nephrotic syndrome (INS). The mean ages of the two groups were similar (4.9 +/- 1.6 vs. 4.6 +/- 2.6 years). Serum albumin levels were similar in both groups, but serum cholesterol was significantly reduced in children with HBV-MN compared to INS. Serum C3 was also significantly depressed in children with HBV-MN compared to INS, but no differences in C4 levels were noted. Serum alanine transaminase as well as aspartate transaminase concentrations were significantly elevated in children with HBV-MN compared to those with INS, suggesting the presence of chronic hepatitis in children with HBV MN. Hepatitis B surface and e antigens were present in serum of all children with HBV-MN, but only 54% had circulating HBV-DNA particles demonstrable in their serum. Serum C3 levels were higher in children with HBV-MN and circulating HBV DNA, compared to those without circulating HBV-DNA. No other serological or biochemical differences occurred between these two groups. Glomerular deposition of IgG and C3 occurred in 91% of children with HBV-MN; but IgM deposition appeared to occur more frequently and with greater intensity in those children positive for circulating HBV-DNA. Antibody to delta antigen was negative in all children with HBV-MN. We conclude that biochemical and serological differences can be identified between HBV-MN and INS.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3041295 TI - Intermediary metabolism and glycemic control in insulin-dependent diabetic uremic patients treated by continuous peritoneal dialysis. AB - The effect on metabolic control and on intermediate metabolism of continuous ambulatory peritoneal dialysis (CAPD) was evaluated in 6 insulin-dependent diabetic uremic patients treated by CAPD, in 6 nondiabetic uremic patients in CAPD and in 6 normal subjects. During the study, 4 dialysis exchanges with 1.36 g/dl dextrose concentration were performed daily; regular insulin was added to the bags in diabetic patients. Our data show a well-controlled mean blood glucose in CAPD diabetic patients by intraperitoneal insulin administration as well as higher insulinemic levels in comparison with those of normal subjects. Plasma lactate and serum glycerol levels were higher and butyrate levels were lower reflecting a continuous ketogenesis inhibition. PMID- 3041296 TI - C5 component: immunopathological index for the activity of IgA nephropathy. PMID- 3041297 TI - Usefulness of serum neopterin in renal transplantation considering the neopterin/creatinine ratio. PMID- 3041298 TI - [Glioblastoma]. PMID- 3041299 TI - [Benign osteoblastoma of the parietal bones]. AB - Benign osteoblastoma accounts for less than 1% of the primary bone tumors and the calvarial lesion is extremely rare. There are only 12 reported cases in the literature as far as we could collect. We have presented a case of benign osteoblastoma originated from the parietal bone. This 9-year-old boy struck his head on the parietal region and noticed the bulging at the same site. A month later he visited to Fukuoka University Hospital because of persistent bulging of the same site. Neurological examinations were normal. Plain skull roentgenogram on Towne view showed radiolucent area in the midline of parietal bones with irregular margins and in tangential view the outer table revealed thinning and expanding. Coronal CT scan demonstrated same abnormality and intact inner table. The tumor was located within the dipole in the bilateral parietal bones and was removed by a simple curettage. The microscopic feature showed numerous osteoblasts, osteoblastic rimming and many multinucleated giant cells. These indicated a typical benign osteoblastoma. He has been doing well one year after the operation without any evidence of recurrence. PMID- 3041300 TI - [Malignant thymoma with intracranial metastases]. AB - Malignant thymomas usually proliferate invasively and rarely metastasize to other organs. Since the metastases occur predominantly to the liver and kidneys, there have been only 16 cases with metastatic spreads of malignant thymomas to the central nervous system reported in the literature. A 59-year-old man was admitted with complaints of dizziness and vomiting. Three years and three months ago, he had been operated upon for mediastinal tumor, which was diagnosed as a predominantly lymphocytic type thymoma, and then followed by irradiation therapy of 3800 rads. The size of the tumor decreased markedly after the irradiation. Nine months after the operation, he complained again of dizziness and vomiting. Computed tomography scans showed a tumorous lesion in the right cerebellar hemisphere, which was thought to be a metastasis from the thymoma. He received radiotherapy of 4000 rads directed to the intracranial metastatic tumor, with the reduction of the tumor size and the relief of symptoms. On the present admission, he had cerebellar signs and symptoms. Neuroradiologically, there was a hypervascular tumor in the left cerebellar hemisphere and a hypovascular one in the right. At operation, a vascular and solid tumor with small necrotic areas were found in the left cerebellum, and a tumor with large liquefied clot within it in the right cerebellum. Pathologically, bilateral cerebellar tumors were confirmed as the metastases from the thymoma in the epithelial type. PMID- 3041301 TI - [Cerebellar ganglioglioma: a case report]. AB - A case of cerebellar ganglioglioma is reported. A 22-year-old female was admitted to the Kurume University Hospital on August 19, 1985, suffering from headache, vomiting and gait disturbance. On admission, neurological examination revealed staggering gait and the right cerebellar ataxia showing dysmetria and dysdiadochokinesis. Mild choked disc in the right fundus was also noted. Plain CT scan showed the low-density area involving the right cerebellar hemisphere and the part of the vermis with internal hydrocephalus. Enhanced CT scan showed the high-density area adjacent to the low-density area suggesting a mural nodule. A vertebral angiogram in the arterial phase showed an expansive lesion in the posterior fossa and the tumor stain, which was also visualized in the venous phase. An emergency suboccipital craniectomy was then performed. With opening the tense dura mater, the cyst formation was noted and 30 ml of xanthochromic fluid was then aspirated. A well demarcated mural nodule was noted in the lateral wall of the cyst. The nodule was then extirpated in toto. The hypertrophy of the cerebellar cortex was not observed. Histologically, the tumor was consisted of nerve fiber, glial fiber and neuronal cells. Its architecture was differed distinctly from that of hamartomatous diffuse hypertrophy of the cerebellar cortex (Lhermitte-Duclos' disease). Immunohistochemically, the neuronal cells revealed positive staining for NSE and S-100, and the glial cells displayed positive staining for GFAP, S-100. The authors reviewed previously reported eleven cases of cerebellar ganglioglioma including the present case. These results showed that cerebellar gangliogliomas have some characteristic clinical features among general intracranial gangliogliomas.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3041302 TI - [Multiple arteriovenous malformations of left parietal lobe and left cerebellar hemisphere with symptomatic trigeminal neuralgia: a case report]. AB - The authors described a case of multiple arteriovenous malformations of the left parietal lobe and the left cerebellar hemisphere, and presented a review of literature. A 42-year-old right-handed man was admitted to our Dept. of Neurosurgery on October 20, 1983, with left facial pain and occipitalgia. He had an episode of subarachnoid hemorrhage ten years ago, however, its etiology was not clear at neurological examination. This brief episode of pain began on the left side of his face about 9 years ago and has been gradually increasing. Although he has been treated with trigeminal nerve blocks several times, relapses were almost always evident within 6 months after those nerve blocks. On admission, there were no abnormal findings at neurological and physical examination. A CT scan with contrast medium infusion revealed two small AVMs on the left parietal region and on the left cerebellar hemisphere. A four-vessels cerebral angiogram confirmed the presence of two AVMs and fenestration of the left vertebral artery. The angiogram also revealed that his severe left trigeminal neuralgia had been caused by elongation of the anterior pontine segment of left SCA which was the main feeder of the cerebellar AVM. Two stages of surgical operations were carried out at two months interval. The first operation was total removal of the cerebellar AVM and microvascular decompression for the left trigeminal nerve. The second was total removal of the left parietal AVM. The postoperative course was uneventful, and angiographically the AVMs completely disappeared. The patient was discharged without any neurological deficits on March 11, 1984.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3041303 TI - [A comparison between stereotaxic operation and conservative therapy for thalamic hemorrhage]. AB - The subjects studied in this report included 17 surgical cases which underwent evacuation of hematoma by means of BRW CT stereotaxic system approximately 14 days after its onset, and 11 non-operated cases. The average age was 65.5 years for the operated and 68.7 years for the non-operated groups. The neurological grades on admission were Grade III or above (slight or mild disturbance of consciousness). The evaluation of the cases was made according to (1) laterality of the hematoma on the left or right, (2) neurological grading, (3) maximum anterio-posterior, lateral diameters, and maximum depth of the hematoma, (4) CT classification (Kanaya, 1981) and ADL (5 grades) at 4.5 months after onset. No correlation was found between ADL and the laterality of the lesion in both groups. As for neurological grading I and ADL on admission, 6/8 cases in the operated and 3/8 in the nonoperated groups recovered to ADL grade 2 or above and 2 bed-ridden cases were included in the latter group. In regard to the extent of the hematoma, the mean ADL was 1.83 in the operated group. While 2.75 in the non operated group where the hematoma was 25 mm or less in its maximum anterio posterior diameter. On CT, there existed a significant difference in ADL at IIb and 4/5 of the operated and 1/4 of the non-operated groups appeared to be ADL 2 or above. Assessment of the activities (mental change, willingness) as well as the muscle strength was performed within 3 days after surgery in operated group and 88.2% and 35.3% improvements were observed, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3041304 TI - The brain in iodine deficiency. AB - Global descriptive, epidemiological studies have established the relation of iodine deficiency to endemic cretinism which, in its fully developed form, is characterized by mental deficiency, deaf mutism and spastic diplegia. However, a second less common variant--myxedematous or hypothyroid cretinism--is characterized by severe hypothyroidism with dwarfism. Mixed forms occur. It has been shown that both conditions can be prevented by correction of the iodine deficiency before pregnancy. Cretinism and development--now termed iodine deficiency disorders (IDD). A number of recently developed animal models establish the effect of severe iodine deficiency on brain development. These include the rat, the marmoset monkey and the sheep. These models are all characterized by the production of severe maternal and fetal hypothyroidism which is associated with effects on the maturation of the cerebral cortex and cerebellum. There was a reduced brain weight with a reduced number of cells as indicated by reduced DNA, a greater density of cells in the cerebral cortex and reduced cell acquisition in the cerebellum. Studies of the mechanisms involved have been carried out in the sheep. The findings reveal significant, though less severe, effects of fetal thyroidectomy (late gestation) and a significant effect of maternal thyroidectomy on brain development in mid-gestation. A combination of maternal and fetal thyroidectomy has similar but more severe effects than iodine deficiency. In the light of current knowledge of the embryology of the brain it is suggested that the critical time for the effect of iodine deficiency is the mid-trimester (14-18 weeks) when the neurons of the cerebral cortex and basal ganglia are formed and could be damaged by the effect of iodine deficiency on maternal thyroid function. There is now recent evidence indicating transfer of maternal thyroxine across the placental barrier early in pregnancy. In this way, neurological cretinism might be produced. Impaired fetal thyroid function would follow in the third trimester and augment the effect of reduced maternal thyroid function. Impaired fetal thyroid function alone could produce the hypothyroid form of cretinism. Further experimental studies, particularly into the postnatal period, are required to substantiate these suggestions. Apart from this, further study of the effects of iodine deficiency on brain development at the subcellular and cellular levels are likely to be most productive. PMID- 3041305 TI - Maintenance of estrous cycle in female rats with anterior or posterior deafferentation of the suprachiasmatic nucleus. AB - The aim of the present study was to determine which of the anterior or posterior efferents of the suprachiasmatic nucleus (SCN) predominantly act for the maintenance of estrous cycle in female rats. A small knife (0.8 mm in radius) was used for anterior (ASD), posterior (PSD) and complete suprachiasmatic deafferentations (CSD). The rats subjected to CSD were divided into two groups, the first group bearing lesions on the medial preoptic nucleus (MPN; group CSD MPN-L) and the other group bearing no lesions on the MPN (group CSD). For reference, anterior hypothalamic deafferentation (AHD) was carried out using a large knife (2.0 mm in radius). All rats with AHD or CSD-MPN-L and some of CSD rats showed persistent estrus, but all animals with ASD or PSD resumed regular estrous cycles after transient irregular periods. Ovaries of AHD, CSD and CSD-MPN L rats contained very few, if any, corpora lutea, while the number of corpora lutea in ASD or PSD rats was not significantly different from that of sham operated controls. Immunohistochemical examination revealed that in AHD rats LHRH immunoreactivity in the median eminence was markedly reduced as compared with that of sham-operated controls. In contrast, in ASD, PSD, CSD and CSD-MPN-L rats, the LHRH immunoreactivity in the median eminence was similar to that of sham operated controls.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3041307 TI - Cumulative indexes. Volumes 11-21. 1982-1987. PMID- 3041306 TI - Relation of the ventromedial nuclei of the hypothalamus to the regulation of renin secretion. AB - During an investigation of the role of the mediobasal hypothalamus in the regulation of renin secretion from the kidneys, we found that lesions of the ventromedial nuclei prevented the increase in plasma renin activity produced by p chloroamphetamine. In the present study, we tested the effects of bilateral electrolytic lesions of the ventromedial nuclei on the increase in plasma renin activity produced in sham-operated rats by immobilization, head-up tilt under inactin anesthesia, and a low-sodium diet. Ventromedial lesions reduced or abolished the plasma renin activity increase to all three stimuli without any change in plasma angiotensinogen. The plasma renin concentration responses to immobilization and a low-sodium diet were also reduced. All these stimuli probably exert their effect by way of the sympathetic nervous system. The data support the hypothesis that the ventromedial nuclei or neural fibers passing through them are important in the renin response to diverse stimuli that act by way of sympathetic discharge. PMID- 3041308 TI - Why don't Broca's aphasics cue themselves? An investigation of phonemic cueing and tip of the tongue information. AB - This study investigates whether Broca's aphasics have the information processing abilities necessary to generate and use their own phonemic cues. Twenty patients were studied; ten benefited from phonemic cues given by the therapist. Phonemic cues were most effective with the patients whose naming was most severely impaired. Six patients could indicate the initial letter of words which they could not produce; three of these patients had no knowledge of any relationship between orthography and phonology, so information about the initial letter must be orthographic and not phonological. Only two patients had any success in giving the sounds of written letters. None of the 20 patients had all three abilities needed to use their own cues: giving the first letter of the name, sounding the letter, and utilizing a phonemic cue. The possibility of relearning letter-to sound correspondences is considered. PMID- 3041309 TI - In vivo voltammetry--present electrodes and methods. PMID- 3041310 TI - Dopaminergic regulation of striatal proenkephalin mRNA and prodynorphin mRNA: contrasting effects of D1 and D2 antagonists. AB - In situ hybridization was used to measure the levels of proenkephalin mRNA and prodynorphin mRNA in regions of rat striatum and nucleus accumbens. Chronic administration of haloperidol (2.4 mg/kg/day for 7 days) increased the levels of proenkephalin mRNA in both striatum and nucleus accumbens. In contrast, the levels of prodynorphin mRNA were not significantly affected in any region. Chronic administration of the D1 antagonist SCH 23390 (2.4 mg/kg/day for 7 days) decreased the striatal content of proenkephalin mRNA. A similar effect was seen in the lateral nucleus accumbens. The levels of prodynorphin mRNA were unaffected by SCH 23390 treatment in all regions examined. These results suggest that there is no major tonic dopaminergic regulation of prodynorphin synthesis in the basal ganglia. However, it appears that there is a tonic suppression, via D2 receptors, and a tonic enhancement, via D1 receptors, of proenkephalin synthesis, in the striatum and nucleus accumbens. PMID- 3041311 TI - Long-term culture of human fetal spinal cord neurons: morphological, immunocytochemical and electrophysiological characteristics. AB - Cultures were prepared from ventral spinal cord tissue from 8-11-week gestational human fetuses and grown for a period of up to 6 months. These cultures were studied by morphological, immunocytochemical and intracellular electrophysiological techniques. From 2 weeks in vitro and onward, small bipolar cells were found in outgrowths of spinal cord explants and were identified as neurons by positive immunoreactions with an antibody specific for neurofilament protein. In addition, a large population of glial fibrillary acidic protein positive astrocytes and a smaller number of galactocerebroside-positive oligodendrocytes were recognized in these cultures. The development of synaptic terminals was also studied by electron microscopy. The first appearance of synaptic terminal was found in a 3-week culture and was an axo-dendritic synapse. During the next 2 months, there was a steady increase in number and structural maturation of synaptic profiles. In addition to axo-dendritic synapses, which were most common, axo-somatic and axo-axonic synapses were demonstrated. After 3 months in culture, the occurrence of large neurons possessing the characteristic features of mature neurons was also noted. Although the occurrence of oligodendrocytes in these cultures was confirmed, no myelination of axons was demonstrated by electron microscopy. Intracellular recordings were obtained from the cultured spinal cord cells, and these cells were identified clearly as neurons by their action potential responses to depolarizing current pulses. The average input resistance of these neurons was 31 M omega with resting membrane potential of -52 +/- 2.3 mV. PMID- 3041312 TI - Clinical significance of types of cerebellar amyloid plaques in human spongiform encephalopathies. AB - We report three patients with both spongiform encephalopathy and cerebellar amyloid plaques; one showed kuru-like plaques and was diagnosed as having Creutzfeldt-Jakob disease (CJD), and two had multicentric plaques and were diagnosed as having Gerstmann-Straussler-Scheinker disease (GSSD). Evaluation of these cases and review of others previously reported suggests a clinicopathologic correlation between type of cerebellar plaque and neurologic clinical course. CJD patients who showed kuru-like plaques generally had disease with early onset (average age, 49.1 years) and long duration (average, 34 months), as compared with CJD patients without kuru-like plaques. GSSD patients usually had multicentric cerebellar plaques, and cases were usually familial, had early age of onset (average, 42.7 years), and were of long duration (average, 73 months). Myoclonus was infrequent in GSSD patients and pathologically spongiform change was minimal; spinal tract degeneration was common. PMID- 3041313 TI - Glucose metabolism alterations in Friedreich's ataxia. AB - We have characterized the abnormalities of glucose metabolism associated with Friedreich's ataxia (FA) by studying plasma glucose, insulin, growth hormone (GH), and glucagon before and after an oral glucose tolerance test (OGTT), an IV glucose load, and an IV arginine load, in 21 patients and in controls. Twelve patients were normotolerant (NT) to glucose, five glucose-intolerant (IT), and four diabetic (DM). Insulin secretion of IT patients was increased and delayed during OGTT. Interestingly, the insulin release during arginine load was significantly decreased in NT and IT as well as in DM patients. The GH response to OGTT was altered in IT patients. Plasma glucagon after an arginine load was significantly higher in patients than in controls. The results indicate that FA is associated with insulin resistance, beta-cell deficiency, and type I diabetes. These alterations might be genetically linked or metabolically related to the primary defect in FA. Their interplay or independent effects are responsible for abnormalities of glucose metabolism in FA. PMID- 3041314 TI - [Retroperitoneal tumors. Problems of diagnosis and therapy]. PMID- 3041315 TI - [Current status of intrathoracic goiter. A review of 58 cases]. PMID- 3041316 TI - [Splenic cysts in childhood. Presentation of 4 cases and review of the literature]. PMID- 3041318 TI - [Clinical experience with nimesulide in gynecologic inflammatory pathology]. PMID- 3041317 TI - [Colorectal cancer and cholelithiasis. Our critical review]. PMID- 3041319 TI - [Calcium antagonists in obstetrics]. PMID- 3041320 TI - Maximal activation of the human diaphragm but not inspiratory intercostal muscles during static inspiratory efforts. AB - It is widely held that transdiaphragmatic pressure is a reliable index of the extent of central activation of the diaphragm but the maximal voluntary transdiaphragmatic pressure is lower during inspiratory than expulsive efforts. To determine whether the diaphragm is fully activated during the two manoeuvres supramaximal stimuli were delivered to both phrenic nerves during maximal efforts. No discernible twitch was evoked during 30-55% of attempted maximal efforts with either voluntary manoeuvre. Thus the difference in maximal transdiaphragmatic pressure between the manoeuvres must reflect changes in chest wall geometry or mechanics rather than in the phrenic motor outflow. Inspiratory intercostal muscle activity was consistently submaximal during maximal inspiratory efforts. PMID- 3041321 TI - A Veterans Administration dental service. PMID- 3041322 TI - [The role of the external carotid artery in the blood supply of the eye from the anatomical and clinical viewpoint]. PMID- 3041324 TI - [Effectiveness of ultrasonic echography in the examination of patients before vitrectomy]. PMID- 3041323 TI - [Results of the surgical treatment of eye injuries complicated by hemophthalmos, using thrombin]. PMID- 3041325 TI - [Biometric parameters of the eyes in young children with congenital glaucoma and their dynamics in long-term observation]. PMID- 3041326 TI - [Clinical aspects of hypophyseal metastases]. AB - Metastatic tumors within the pituitary gland are rare. The incidence--reported by series of autopsies--differs from 1.8-12% of metastatic tumors. Most of the primary tumors are breast cancers, followed by cancers of the gastro-intestinal system. Only 20% of these metastases are clinically diagnosed. Diabetes insipidus is the main symptom, correlating with the greater incidence of metastatic lesions in the posterior lobe. The unusual features of a reported case are discussed. In this case the dysfunction of cranial nerves passing the sella turcica led to the diagnosis. Therapy with Flutamid initiated a remission. Survival of 18 months is remarkably long. PMID- 3041327 TI - [Randomized double-blind study of the analgesic effect of caerulein and morphine in chronic tumor pain]. AB - Caerulein (CRL) (5 micrograms i.m.) and morphine (10 mg i.m.) have been tested for their analgesic activity in a double blind randomized study in a total of 36 patients with medium to severe tumor pains. A decrease of more than 20 mm on a visual analogue scale (VAS) was taken as a criterion for successful therapy. This was the case in 67% of the patients treated with morphine and 50% of those treated with CRL. This difference is statistically not significant, but CRL has significantly fewer side-effects than morphine. The present data do not permit a definitive judgement on the value of CRL in the treatment of tumor pains. Further studies with more patients and different doses and administration routes are warranted. PMID- 3041328 TI - [CT kinetics of intratumor liposome deposits]. AB - CT follow-up studies of liposome-entrapped metrizamide after intraneoplastic injection into neurogenic s.c. rat tumors were performed. By closely resembling clinical examination conditions, the experimental design has proven suitable in determining the in vivo kinetics of these interstitial liposome deposits. When compared to free metrizamide which may be considered an analogue of water-soluble chemotherapeutics, the encapsulation of metrizamide in liposomes resulted in a retarded decline of the contrast enhancement. Diffusion of liposomes could not be detected and the X-ray attenuation values measured within the liposome deposits continuously decreased with time for both types of liposomes. In the case of multilamellar vesicles, this significantly corresponded to a zero order kinetics with a mean halflife of 300 h. An initial increment in the X-ray attenuation of the liposome deposits might be due to the interstitial absorption of the water component of the liposome-dispersion. Because of the pronounced retardation effect of multilamellar liposomes resulting in a 140-fold prolongation of the interstitial retention time of metrizamide and due to their release kinetics these vesicles may be an appropriate carrier system for a local interstitial chemotherapy modality. Small unilamellar vesicles having an interstitial half life of 14 h may be used as a faster component of a composed therapy system. PMID- 3041330 TI - Proof: an elusive ideal. PMID- 3041329 TI - [Pathophysiology of hyperprolactinemia in breast cancer]. AB - To characterize the prolactin secretion in human breast cancer, plasma prolactin levels were measured in 514 patients with breast cancer in long term follow-up studies. In hyperprolactinemic patients suppression and stimulation tests were performed and the 24-h secretion profile was recorded. Tissue extracts and sera of hyperprolactinemic breast cancer patients were incubated with cultured pituitary cells in vitro to detect a prolactin releasing activity in these specimens. 44% of breast cancer patients developed hyperprolactinemia in the course of the disease. In 35% of measurements hyperprolactinemia was induced by non tumor related causes, e.g. prolactin-stimulating drugs, surgery, uremia, prolactinoma. Excluding such influences on the prolactin level, hyperprolactinemia over 1,000 mU/l was almost only found in patients with progressive metastatic disease. In these patients hyperprolactinemia was associated with tumor load, but not correlated to BSR, CEA or prognostic factors. Hyperprolactinemia in breast cancer was not of paraneoplastic origin. No prolactin-releasing activity was detected in tumor tissue and sera of hyperprolactinemic breast cancer patients. PMID- 3041332 TI - Corneal decompensation following acute angle-closure glaucoma. AB - Ten eyes of nine patients with endothelial dystrophy requiring iridectomy for acute angle-closure glaucoma developed corneal edema sufficient to require penetrating keratoplasty. Because of the proximity of iris to cornea, simultaneous lens extraction was carried out to prevent malignant glaucoma. Penetrating keratoplasty using techniques presented here provided improved visual acuity and control of glaucoma in all cases. PMID- 3041331 TI - Tarsoconjunctival grafts for upper eyelid cicatricial entropion. AB - Many surgical procedures have been described for correction of cicatricial entropion of the upper eyelid. However, many of them fail to address the altered anatomy responsible for cicatricial entropion, which may lead to excessive scarring, eyelid margin malposition, or blepharoptosis. Tarsoconjunctival grafts provide a strong and permanent buttress to correct the scarring of the posterior eyelid margin that characterizes cicatricial entropion. They provide a smooth mucosal surface to interface with the corneal tear film. Depending on the circumstances, we use free ipsilateral, free contralateral, or sliding "bucket handle" tarsoconjunctival grafting. Mucosal grafts are rarely needed. Blepharoptosis is avoided by conservative dissection in the supratarsal space, sparing most of the attachments of the levator aponeurosis. PMID- 3041333 TI - Corneal traction suture. PMID- 3041334 TI - Disparate diameter technique. PMID- 3041335 TI - Treatment of oro-facial herpes simplex infections with acyclovir: a review. AB - The treatment of herpes simplex virus (HSV) infections in the past has been largely unsuccessful. Introduction of the drug acyclovir has been a positive development. Acyclovir has been extensively studied in the treatment of a a variety of HSV infections in immunocompromised patients and in otherwise healthy patients. The results have shown it to effectively inhibit HSV replication but to have no effect in preventing or eliminating the latent state of the virus. It has been shown to be very effective in certain instances and not so effective in others. PMID- 3041336 TI - Performance of a hydroxypropyl cellulose film former in normal and ulcerated oral mucosa. AB - Although oral ulcers are commonly encountered in clinical dental and medical practice, current therapeutic options with respect to pain relief are limited. This study evaluated the mucosal binding characteristics and the pain relief and protection properties of Zilactin, a hydroxypropyl cellulose film former. In 12 healthy volunteers, the mean duration of mucosal adherence of Zilactin was 3.92 hours, which was significantly longer than that of a widely available topical preparation (1.38 hours, p = 0.0001). When evaluated in 20 subjects with recurrent aphthous ulcerations, Zilactin demonstrated the ability to significantly decrease ulcer pain over a 4-hour period when compared with the subjects' individual standardized level of initial discomfort, which was used as an internal control for all subsequent steps. Challenge with an irritating citrus beverage was made before and after application of the medication. A significant difference in premedication and postmedication sensitivity was shown (p = 0.0001), indicating that the film provides protection, as well as pain relief. As a result of its unique and tenacious film-forming characteristics, this agent is exceptionally effective in the treatment of oral mucosal ulcerations. Greater awareness on the part of physicians and dentists may lead to the development of additional applications. PMID- 3041337 TI - Eosinophilic granuloma. AB - Eosinophilic granuloma is said to be the most benign disorder of the triad commonly known as histiocytosis X. There has been, and still is, confusion about the terminology describing this entity. This article reviews the literature on histiocytosis X, with particular emphasis on eosinophilic granuloma and its oral manifestations. The case of a 22-month-old male child with an eosinophilic granuloma of the right mandible is presented. The diagnostic and treatment methods are reviewed and discussed. During the 4 years that the case has been followed there has been progressive healing of the lesion, but other manifestations of the disease process and of the surgical treatment have become evident. The need for close dental as well as medical follow-up in cases involving the dental structures is stressed. PMID- 3041338 TI - Lingual metastasis of alveolar soft-part sarcoma. AB - A patient with lingual metastasis of alveolar soft-part sarcoma, which originated in the brachioradial muscle, is presented and the literature, is reviewed. PMID- 3041339 TI - The flare-up phenomenon in endodontics: a clinical perspective and review. AB - The acute endodontic cellulitis exacerbation, which can be potentially fatal, is a definitive entity in endodontic flare-ups. Aerobic microbes, particularly streptococci, are the predominant causative microbes isolated. There was a noticeable absence of obligate anaerobes. This is significant for the selection of an antibiotic for therapy. Treatment parameters were presented. An endodontic cellulitis exacerbation is most unlikely with obligate anaerobes. An endodontic flare-up perspective was attempted with some clinical parameters. The proponents of routine one-visit endodontic treatment with prophylactic drugs to prevent cellulitis exacerbations do not appear to offer any advantage to the more traditional approaches to endodontic treatment of the patient, which may be more beneficial. PMID- 3041340 TI - Multiple idiopathic external root resorption. A case report. AB - An unusual case of multiple idiopathic external apical root resorption affecting all four quadrants, in a patient with a history of narcotic intravenous drug addiction and liver disease, is reported. The literature relating to a systemic etiology for external root resorption is briefly reviewed. PMID- 3041342 TI - Periostitis ossificans versus Garre's osteomyelitis. Part I. What did Garre really say? AB - In 1893, C. Garre published an article dealing with the manifestations of acute osteomyelitis. Since then, his name has been associated with diseases such as Garre's osteomyelitis, chronic sclerosing osteomyelitis, and periostitis ossificans, among others. Scrutiny of a translated version of the original article reveals that Garre was not responsible for the description of the disease that now bears his name. PMID- 3041341 TI - Film-holding instruments for intraoral subtraction radiography. AB - Clinical application of digital subtraction radiography is limited by the reproducibility in the orientation of the x-ray source, image receptor, and object. In this study, eight dental intraoral film alignment instruments (including five replicates of each) were tested for accuracy in repositioning over a period of 6 months. Each instrument was made by adhering one of six impression materials (including acrylics, compounds, and elastics) onto the bite blocks of commercially available alignment instruments. The dimensional accuracy and reproducibility of the orientation of the x-ray source with respect to the object were determined over time for each instrument by measuring the horizontal and vertical angulation change in the position of a buccally placed marker on a dried mandible with a lingually placed film grid. Nine measurements were made during the 6-month test period. The most reproducible instrument was the combination of Regisil, an elastic impression material, and a Rinn XCP bite block. This combination yielded a mean absolute horizontal angulation error of 1.34 degrees +/- 0.63 and a mean absolute vertical angulation error of 2.04 degrees +/- 0.82, yielding a total angulation error of 2.44 degrees +/- 1.16. This was within the acceptable range of accuracy needed to produce diagnostically useful information when digital subtraction radiography is used. PMID- 3041343 TI - [40 years of Austrian Nursing Federation 1948 to 1988]. PMID- 3041344 TI - Cloning and expression of chicken p54c-ets cDNAs: the first p54c-ets coding exon is located into the 40.0 kbp genomic domain unrelated to v-ets. AB - We have isolated cDNA clones of chicken c-ets mRNA the longest of which, designated pCk E54A, contained approximately 2.0 kb of a c-ets mRNA species. Nucleotide sequencing of this clone revealed a single long open reading frame, extending from the first ATG codon (nucleotide +1) to a TGA termination codon at nucleotide 1324. The predicted translation product contains 441 amino acid residues and its molecular weight is 48 kd. Expression in COS-1 cells of this clone resulted in the synthesis of polypeptides immunologically indistinguishable from the authentic p54c-ets after one-dimensional gel electrophoresis. Comparison of the nucleotide sequence of this cDNA to that of v-ets of avian acute leukemia virus E26 showed that both sequences are almost colinear with the exception of five point mutations but present striking differences in their 5' and 3' parts. 79 nucleotides downstream of the first ATG codon in c-ets cDNA are not found in the 5' part of v-ets where they are replaced by 223 different nucleotides. The 3' parts of v-ets and the coding region of the chicken c-ets cDNAs are also different: the last 13 codons of the cDNA are replaced by 16 different codons in v-ets. Thus our results precisely define the structural differences between the ets encoded domain of E26 viral transforming protein (P135 gag-myb-ets) and the normal cellular protein p54c-ets expressed at high levels in chicken thymocytes and bursal lymphocytes. They also suggest the possibility of alternative splicing of different 5' exons to a common set of 3' exons. PMID- 3041345 TI - Differential developmental expression of cellular yes and cellular src proteins in cerebellum. AB - To identify the specific areas of the brain that express c-yes and c-src proteins, we examined chicken brains dissected from two-week-old birds using an immune complex kinase assay and an immune blot analysis. Highest levels of both proto-oncogene proteins were found in the cerebellum, whereas other parts of the brain, including telencephalon, diencephalon, mesencephalon and spinal cord, showed three- to six-times lower levels. Relatively low levels of the two proteins were detected in pineal body and pituitary. When the cerebellum was further dissected into three layers, molecular, granular and fibrous, both the c yes and c-src proteins were found to be concentrated in the molecular layer and, to a lesser degree, also in the granular layer. In cerebellum and in chicken embryo fibroblasts the c-yes protein was predominantly associated with the membrane fraction, and in chicken embryo fibroblasts c-yes was labeled with radioactive myristic acid. Adult cerebellum showed a two- to three-fold increase in the c-yes protein level over that detected in embryonal cerebellum. Conversely, c-src expression in the embryonic cerebellum was relatively high and it was diminished in the adult cerebellum. Differential developmental expression of c-yes and c-src proteins in cerebellum suggests that these proteins fulfill different functions or different aspects of the same function. PMID- 3041346 TI - Complementary DNA clones of chicken proto-oncogene c-ets: sequence divergence from the viral oncogene v-ets. AB - The avian acute leukemia virus E26 induces erythroblastosis and myeloblastosis in chickens. The oncogene of this virus includes sequences derived from the cellular gene designated c-ets, which is normally expressed in lymphoid cells and whose product is a protein of apparent molecular weight ca. 54,000 daltons. Complementary DNA clones representing the major transcript of the chicken c-ets proto-oncogene were isolated from a spleen cell library. Sequence analysis of the cDNA revealed that it contains an open reading frame encoding a polypeptide of 441 amino acids with a molecular weight of 49,932 daltons. This open reading frame can be transcribed and translated in vitro into a 50 kd protein that is specifically immunoprecipitated with antiserum to the v-ets oncogene product. Within the central region of homology between c-ets and v-ets, there are only 5 nucleotide substitutions resulting in 4 amino acid changes. However, coding sequences at the 5' and 3' ends of the v-ets oncogene and the chicken c-ets cDNA differ from one another. These changes may be responsible for the differential functions of c-ets and v-ets in cells of different hematopoietic lineages and may account for the pathogenic properties of the v-ets oncogene. PMID- 3041347 TI - Post-translational alterations of the tyrosine kinase p56lck in response to activators of protein kinase C. AB - We have found in different human cells of lymphoid and non-lymphoid origin that the 56 kilodalton (kDa) lck protein is rapidly converted to a product migrating at approximately 60 kDa (designated p60lck) in response to the phorbol ester 4 alpha-phorbol 12 beta-myristate (PMA) as well as the diacylglycerol analogue 1,2 dioctanoyl-sn-glycerol (diC8). This conversion is associated with an increase in serine phosphorylation within the amino-terminal 18 kDa portion of the lck protein. The serine phosphorylation modification and diminished electrophoretic mobility of the lck protein appear to be completely reversible within 60 min following treatment with diC8. The changes in p56lck phosphorylation and gel mobility in response to activators of protein kinase C are also associated with a small but reproducible decrease in the ability of the lck protein to be phosphorylated in immune complex kinase assays. While these alterations of the lck gene product may play an important role in antigen-mediated activation of T lymphocytes, we demonstrated that they can also be induced independently of T cell activation suggesting that they are not necessarily implicative of this process. PMID- 3041348 TI - Imaging modalities for the study of the paranasal sinuses and nasopharynx. AB - Disease entities affecting the paranasal sinuses and nasopharynx are now evaluated primarily by computed tomography (CT) and magnetic resonance imaging (MRI). Cortical bone is best seen by CT, particularly with thin sections and high resolution bone mode. Bone marrow and various soft-tissue structures are best evaluated by MRI. This article is a demonstration of the normal anatomy of the area as depicted by both CT and MRI. PMID- 3041349 TI - Inflammatory diseases of the paranasal sinuses and mucoceles. AB - Inflammatory diseases of the paranasal sinuses are commonly encountered. In addition to the clinical evaluation, paranasal sinus films are often included for the diagnosis and follow-up. Complications of such infections and mucoceles are best evaluated by CT and MRI. PMID- 3041350 TI - Tumors of the paranasal sinuses. AB - Benign and malignant tumors, including polyps, represent conditions that occur frequently enough to be considered in the differential diagnosis of sinus disease. The locations and extent of these lesions (especially malignant tumors) should be evaluated by CT and MRI. This is especially indicated when malignant lesions extend outside the confines of the paranasal sinuses into adjacent areas, such as the orbit, intracranial cavity, and parapharnygeal space. PMID- 3041351 TI - Traumatic injuries of the paranasal sinuses. AB - High-resolution imaging, based on CT, has become the expected standard of imaging in severe sinofacial trauma. Imaging must include both bone and soft-tissue detail. Intracranial complications such as hematoma, contusion, and dural tear must be noted and followed appropriately. Early orbital assessment must be included to allow surgical decompression of hematoma or fracture reduction before irreversible changes to the visual pathway occur. Clinical assessment and initial, limited plain films offer an invaluable overview to set up priorities in resuscitation and subsequent direction for more detailed assessment by higher resolution imaging. Complex facial fractures in noncontiguous structures are increasingly noted with high-velocity trauma. Open communication between clinician and radiologist should prevent only partial assessment of the true extent of involvement. Increasing use of CT (and possibly MRI in the near future) to follow persisting post-reduction complications (Fig. 14)--whether altered position of bone grafts or implants, ocular motility disorders and enophthalmos, or sinus obliteration or ablation--has resulted in the further need for the clinician and radiologist to understand each other's capabilities, in order to offer the patient maximum benefit from his or her imaging referral. PMID- 3041352 TI - Imaging modalities for the evaluation of neck pathology. AB - CT and MRI are the primary imaging modalities used to evaluate patients with neck pathology. The lack of good surface coils is a major limitation in the use of MRI in the evaluation of the portion of the neck between the hyoid bone and the thoracic inlet. Most radiologists prefer to use CT for the evaluation of pathology in the infrahyoid portion of the neck. PMID- 3041353 TI - Assessment of salivary gland pathology. AB - At this institution, inflammatory problems and calculous disease are evaluated by plain films or sialography. CT can be used for stone evaluation but is not usually necessary. Although CT has been the mainstay of tumor evaluation, magnetic resonance imaging has several very definite advantages and is likely to supercede CT as the procedure of choice. PMID- 3041355 TI - Immunoglobulin therapy of neonatal group B streptococcal infections: an overview. AB - Group B streptococci (GBS) are a major cause of sepsis and meningitis in newborn babies. Neonatal GBS infections are often rapidly progressive, suggesting that the immunity to GBS is deficient. Studies have shown that opsonic antibody is required for efficient phagocytosis and killing of GBS, and neonatal GBS infections have been associated with diminished levels of anti-GBS antibody. Intravenous immunoglobulin (IVIG) has been shown to provide protective immunity in experimental GBS infection models. However, lot to lot variation in opsonic antibody levels occurs in standard IVIG preparations. Recently hyperimmune anti GBS IVIG has been prepared with high levels of opsonic and protective antibody to GBS. Hyperimmune IVIG preparations will allow physicians to give higher quantities of specific anti-GBS antibody without having to administer large fluid volumes or large amounts of nonspecific immunoglobulin. In addition specific immunoglobulin preparations will ensure that the IVIG contains reliable antibacterial activity. Although human studies are limited they suggest that IVIG therapy in neonates may be safe and effective in treating neonatal sepsis. However, further studies are necessary to determine the role of IVIG in preventing or treating neonatal infections. PMID- 3041354 TI - Respiratory syncytial virus infections and intravenous gamma-globulins. AB - The respiratory syncytial virus (RSV) is a common cause of bronchiolitis and pneumonia in infants and young children. Throughout the world annual RSV epidemics result in numerous hospitalizations, substantial morbidity and some mortality. Until the recent introduction of ribavirin only supportive therapy has been available for treating these infections. The development of animal models of RSV infection and the observation that some lots of immunoglobulin prepared for intravenous administration contained substantial RSV-neutralization antibody titers, prompted a series of studies examining the safety and efficacy of immunoglobulin prepared for intravenous administration in the prophylaxis and treatment of RSV infections. This discussion will review our published, or soon to be published, studies on the use of Sandoglobulin for both immunoprophylaxis and immunotherapy of RSV infections in cotton rats. It will summarize studies utilizing both parenteral and topical (tracheal) Sandoglobulin therapy for RSV infections in owl monkeys. Finally the results of a small double blind trial of parenteral albumin or Sandoglobulin in the therapy of RSV bronchiolitis and/or pneumonia in hospitalized children will be reviewed. The data show that immunoprophylaxis and immunotherapy of RSV infections in laboratory animals was well-tolerated, was safe and induced highly significant reductions in RSV shedding from the lower respiratory tract. Further, immunotherapy of RSV infections in children was also well-tolerated, induced no short or long term evidence of toxicity or injury and caused significant improvements in oxygenation and reductions in RSV shed from the respiratory tract.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3041356 TI - Immunoglobulin subclass deficiency. AB - IgG subclass deficiency was first noted in 1968. Subnormal levels of one or two, occasionally three IgG subclasses may be relatively common. It has not been determined, however, at what level below the normal range the IgG subclass deficiency is of clinical relevance. It remains important to clarify this point because certain subclass deficiencies may be without relevance of their own. Because patients with decreases of various IgG subclasses often present with a number of diseases, the low immunoglobulin levels may signify the presence of other abnormalities of more biologic significance. IgG subclass deficiency has been noted in about 25% of patients with well-defined food allergy and in patients with asthma, diabetes mellitus, Henoch-Schonlein's purpura, Bechterew's disease, intractable epilepsy of childhood, Friedreich's ataxia and autoimmune cytopenias. Most commonly they have increased frequency of infections especially in the respiratory tract, including sinusitis, otitis media and bronchopneumonia, but also osteomyelitis, meningitis, septicemia and various skin infections. Low levels of various subclasses have been noted in connection with other immunodeficiencies such as ataxia-telangiectasia. In common variable immunodeficiency there is an obvious imbalance in the IgG subclasses. Furthermore IgG subclass deficiency can be seen in relatives of patients with common variable immunodeficiency and in IgA deficiency. They also occur in relatives of patients with systemic lupus erythematosus, diabetes mellitus type 1 and C2 deficiency. In a few cases of subclass deficiency gene deletions have been shown. Subnormal levels of IgG subclasses make a remarkable change in sex distribution around puberty from 3/1 in boys and girls to the reverse sex ratio among adults.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3041357 TI - Infections and immunodeficiency in bone marrow transplantation. AB - After allogeneic bone marrow transplantation certain patterns of infectious complications emerge that follow the clinical course, are correlated to the immunobiology of transplantation and are almost predictable in their character and expression. The preparative regimen, designed to generate complete aplasia, will be associated with severe and sometimes life-threatening bacterial infections, predominantly with Gram-negative organisms derived from bowel flora, but also Gram-positive skin saprophytes. In this early aplastic phase, life threatening viral infections are less common, consisting mainly of herpes simplex and possibly Epstein-Barr stomatitis and BK papovavirus cystitis. Systemic infections with invasive filamentous fungi are rare and are seen only when the induced aplasia is markedly prolonged. Once early marrow recovery has been achieved, systemic infections will generally disappear unless acute graft-vs. host disease develops. This complication, which will lead to the breakdown of natural barriers such as skin and gastrointestinal epithelium and the marked impairment of all systemic defense mechanisms, can cause polymicrobial infections as well as set the stage for life-threatening viral infections. Such opportunistic viral infections, leading to either interstitial pneumonia or hemorrhagic gastroenteritis, are the major threat in the early recovery phase after engraftment has taken place. Usually caused by cytomegalovirus and rotavirus, respectively, these infections are the primary expression of the severe combined immunodeficiency post transplant, statistically associated with the presence of acute graft-vs.-host disease and amenable to immunologic manipulations. With the recovery of cellular and humoral immune function derived from transplanted donor lymphoid cells, the third phase of infectious complications is reached, covering 3 months to 2 years post grafting.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3041358 TI - High dose intravenous immunoglobulin therapy in burn patients: pharmacokinetics and effects on microbial opsonization and phagocytosis. AB - Depressed serum immunoglobulin levels following severe burns may lead to subsequent infectious complications following such injuries. In a randomized study we administered multiple doses of Sandoglobulin (500 mg/kg) or albumin intravenously to patients with severe burn injuries and closely monitored serum IgG levels. Patients who received IgG therapy had earlier return of normal serum IgG levels compared to control patients; however, control patients attained normal IgG levels during the second postburn week. Serum half-lives of IgG following infusions were remarkably short (means, 47 hours for infusions within 3 days of injury and 154 hours for infusions in the third postburn week); Sandoglobulin has been reported to have approximately a 21-day half-life in normal individuals. We also measured the opsonic capacity of postburn serum, using fluorescein-labeled microbes and flow cytometry; we identified postburn opsonic defects with certain of the organisms as late as 15 days postinjury, even though serum IgG levels had normalized. These defects were corrected by the in vitro addition of Sandoglobulin to the incubation mixture. PMID- 3041359 TI - Antibody deficiency syndromes and novel immunodeficiencies. AB - Antibody deficiency syndromes can be quantitative or qualitative. The major categories of antibody deficiency syndrome are: (1) X-linked agammaglobulinemia, involving the maturation arrest in the development of the B cells; (2) transient agammaglobulinemia, which affects both sexes is often associated with defective T helper function for immunoglobulin production; (3) acquired agammaglobulinemia, a heterogeneous disorder caused by a primary B cell defect, absence of B cells, presence of B cells but with an activation defect, failure of helper factor production by T cells or increase in suppressor cells; (4) IgG2 and IgG3 subclass deficiencies, causing significant and recurrent infections and, with IgG2, a significant impairment of the ability to respond to carbohydrate antigens such as Haemophilus influenzae, pneumococcus and meningococcus; (5) qualitative antibody deficiency syndrome in the response to carbohydrate antigens, presenting as recurrent infection and involving the inability of the patient to respond to immunization with polysaccharide antigens such as Haemophilus influenzae type b; (6) antibody deficiency states associated with T cell dysfunction. Impairment of T cell function, which is required for B cell activation, presents often as antibody deficiency syndrome. In these cases, total gamma-globulin level is normal, but the quality of the antibody is very poor.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3041360 TI - Human immunodeficiency virus infection in children: nature of immunodeficiency, clinical spectrum and management. AB - The causative agent of acquired immunodeficiency syndrome is a retrovirus, human T lymphotropic virus type III/lymphadenopathy-associated virus, now known as human immunodeficiency virus (HIV). Infection of children with HIV results in a wide spectrum of clinical manifestations, ranging from asymptomatic to symptomatic, with the severest disease forms including neurologic deterioration, opportunistic infections and malignancy. This virus infects preferentially T cells bearing the CD4 receptors and also seems to exhibit preference for the central nervous system. The predominant route of infection in children is transplacental, and most affected children are infected at the time of birth. For women who give birth to infants with congenital infection with HIV, the main risk factor is intravenous drug abuse; a smaller percentage of these women acquire the infection via sexual contact and a few are infected via blood transfusions. Estimates for the incidence of transmission of the virus from an infected mother to her offspring vary from about 20 to 70%. Infection in most children and adults is documented by serologic testing, inasmuch as almost all infected people are HIV antibody-positive. Mothers of congenitally affected children are always HIV antibody-positive and also frequently have immune abnormalities. Women who give birth to infected children may, however, be asymptomatic in 50% of instances or more. Because antibodies to HIV are predominantly of the IgG class, they cross the placenta. All infants born to infected women therefore acquire passively transferred antibodies to HIV irrespective of whether or not the infants are infected with the virus itself. These passively transferred antibodies may sometimes persist for as long as 15 months. Thus in infants and children under 15 months of age in the absence of symptoms, the only definitive way to establish diagnosis is by viral isolation or viral antigen detection. Clinically the HIV infected children can be divided into two groups, symptomatic and asymptomatic. Among the symptomatic group the main diagnostic specific features are: (1) opportunistic infection, e.g. with Pneumocystis carinii pneumonia; (2) interstitial pneumonitis with respiratory distress resulting from lymphocytic interstitial pneumonitis; (3) microcephaly and other neurologic abnormalities; (4) recurrent bacterial infections.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3041361 TI - Changes in life-style after liver transplantation. AB - Sixty-five pediatric patients who received liver transplants between May 1981 and May 1984 were observed for as many as 5 years and examined for changes in life style. Children were less frequently hospitalized, spent less time hospitalized, required fewer medications, and generally had excellent liver and renal function after hepatic transplantation as compared with their pretransplantation status. Most children were in age-appropriate and standard school classes or were only 1 year behind. Cognitive abilities remained unchanged. Children improved in gross motor function and patients' behavior significantly improved according to parents' perceptions. Enuresis was more prevalent, however, than in the population of children who had not received liver transplants. Parental divorce rates were no greater than those reported for other families with chronically ill children. Overall, objective changes in life-style as well as parents' perceptions of behavior of children appear to be improved after liver transplantation. PMID- 3041362 TI - Toxoplasmosis: maternal and pediatric findings in 23,000 pregnancies. AB - An analysis of the antibody titers to toxoplasmosis for 22,845 pregnant women in the Collaborative Perinatal Project was conducted in relation to clinical and laboratory findings in the mothers and children through 7 years of age. More than 900 observations were considered for each mother and child. The major findings were in the children and included a predicted doubling in the frequency of deafness among children born to women with antibody to toxoplasmosis, a predicted 60% increase in microcephaly, and a 30% increase in low IQ (less than 70) in association with the presence of high maternal antibody titer (256 to 512) to toxoplasma. A serologically defined high-risk group of mothers was identified on the basis of high indirect hemagglutination antibody levels or seroconversions and increased IgM toxoplasma antibody levels (indirect fluorescent antibody greater than or equal to 32, enzyme-linked immunosorbent assay greater than or equal to 0.7). Of the 15 pregnancies in this group, two children had congenital toxoplasmosis and three were stillborn. PMID- 3041363 TI - Intrapartum asphyxia and cerebral palsy. AB - Signs of presumed hypoxia/asphyxia of the fetus are not uncommon and can be detected during labor, in the delivery room, and during the early neonatal period. Virtually no single sign or symptom has sufficient correlation to enable prediction of later cerebral palsy with a reasonable degree of medical certainty. To attribute cerebral palsy to prior asphyxia with reasonable certainty, there must be evidence that a substantial hypoxic injury occurred and that a sequence of events ensued which would prove the clinical impact of that hypoxic insult. Few cases of cerebral palsy meet these criteria. PMID- 3041364 TI - Anencephalic infants as organ donors. AB - Transplantation technology has been refined in recent years and now offers hope to pediatric patients with a variety of end stage disease processes. The lack of available donors for the smallest potential organ recipients has led to the suggestion that anencephalic infants be used as organ donors. This suggested policy is contrary to current law and raises fundamental ethical issues relating to the definition of death and the treatment of the dying. The technical issues in the potential organ supply from this source are described and the opposing ethical positions developing in this debate are discussed. PMID- 3041365 TI - Harvesting organs for transplantation from dying anencephalic infants. PMID- 3041366 TI - Contributions to the history of psychology: XLVII. Ignatius Loyola and behavior therapy. PMID- 3041367 TI - Dependence upon high-energy phosphates of the effects of inorganic phosphate on contractile properties in chemically skinned rat cardiac fibres. AB - The effects of inorganic phosphate (Pi) on mechanical properties of Triton X100 treated ventricular fibres have been studied in different substrate conditions. In the presence of both MgATP and phosphocreatine, increasing concentrations of Pi progressively decreased maximal active force, up to 50-60% at 20 mM Pi. The reduction in stiffness was slightly less. These effects appeared nearly independent of the diameter of the preparations. 20 mM Pi decreased Ca sensitivity of the myofilaments and increased the Hill coefficient of the tension/pCa relationship; furthermore, the time constant of tension recovery was decreased from 12.9 to 8.9 ms suggesting that the cycling rate of cross-bridges was increased in the presence of Pi. When MgATP was regenerated by the myofilament bound creatine kinase in the presence of phosphocreatine, Pi was less efficient in decreasing the maximal tension and it weakened the relaxing effect of MgATP upon rigor tension. These effects are related to the inhibition of creatine kinase by Pi. The effects of Pi on maximal force and kinetics of contraction were antagonized by the effects of a decrease in phosphocreatine. The results are discussed in terms of the antagonistic role of Pi increase and phosphocreatine decrease upon contractile properties of myofilaments during hypoxia in heart muscle. PMID- 3041368 TI - Photocleavage of DNA and photofootprinting of E. coli RNA polymerase bound to promoter DNA by azido-9-acridinylamines. AB - The long-wavelength ultraviolet (lambda approximately 420 nm) radiation induced reaction between 6-azido-2-methoxy-9-acridinylamines and supercoiled plasmid DNA results in single strand scissions and formation of covalent adducts (ratio approximately 1:10). By treating azidoacridine-photomodified DNA with piperidine at 90 degrees C, additional strand scissions are observed in a complex sequence dependent manner with an overall preference for T greater than or equal to G greater than C much greater than A. The resulting DNA fragments migrate as 5' phosphates in polyacrylamide gels. Photofootprinting of the binding site of RNA polymerase on promoter DNA is demonstrated with an azido-9-acridinylamino octamethylene-9-aminoacridine. Similar experiments using 9-amino-6-azido-2 methoxyacridine indicate that this reagent recognizes changes in the DNA conformation induced by RNA polymerase binding, in relation to open complex formation. PMID- 3041370 TI - High efficiency transformation of E. coli by high voltage electroporation. AB - E. coli can be transformed to extremely high efficiencies by subjecting a mixture of cells and DNA to brief but intense electrical fields of exponential decay waveform (electroporation). We have obtained 10(9) to 10(10) transformants/micrograms with strains LE392 and DH5 alpha, and plasmids pUC18 and pBR329. The process is highly dependent on two characteristics of the electrical pulse: the electric field strength and the pulse length (RC time constant). The frequency of transformation is a linear function of the DNA concentration over at least six orders of magnitude; and the efficiency of transformation is a function of the cell concentration. Most of the surviving cells are competent with up to 80% transformed at high DNA concentration. The mechanism does not appear to include binding of the DNA to the cells prior to entry. Possible mechanisms are discussed and a simple procedure for the practical use of this technique is presented. PMID- 3041369 TI - Genome organization of the killer plasmid pGK12 from Kluyveromyces lactis. AB - We have determined the entire sequence of the plasmid K2 from Kluyveromyces lactis which is involved in the maintenance of both killer plasmids in the cell. K2 shares many of the characteristics of the smaller killer plasmid K1: high A+T content (74.7%) and very compact genomic organization. K2 contains ten open reading frames. Some of them overlap on different strands and some on the same strand. Northern blotting of K2 transcripts shows that at least eight ORFs are transcribed. Analysis of the predicted aminoacid sequence of ORF2 from K2 reveals homology with the aminoacid sequence of ORF 1 from K1 and with several viral DNA polymerases. The sequence of K2 from Saccaromyces cerevisiae F102-2 was also determined. Only one nucleotide difference was found between the K2 sequence from the two yeasts. This mutation does not change the genome organization of the plasmid and has only minimal effect on the structure of the encoded proteins. PMID- 3041371 TI - Escherichia coli RNase D: sequencing of the rnd structural gene and purification of the overexpressed protein. AB - We have determined the nucleotide sequence of a 1.4-kb-pair fragment of the E. coli chromosome that carries the complete rnd gene encoding RNase D, a putative tRNA processing enzyme. The coding region of rnd extends for a total of 1128 nucleotides beginning at an initiator UUG codon and terminating at a UAA codon, and encodes a 375-amino acid polypeptide of 42,679 daltons, consistent with the known size of RNase D. A rapid purification procedure was developed for isolation of RNase D from strains overexpressing the enzyme. The N-terminal sequence and the amino acid composition of the homogenous protein were in excellent agreement with those derived from the sequence of the rnd gene. PMID- 3041372 TI - Effects of macromolecular crowding on the association of E. coli ribosomal particles. AB - The equilibrium for the binding reaction between the 30 S and 50 S ribosomal subunits of E. coli is shifted towards formation of 70 S ribosomes in the presence of a variety of polymers. The polymers also increase a further interaction between 70 S particles to form the 100 S dimer. The requirement for relatively high concentrations of non-specific polymers indicates that the shifts in equilibria arise from excluded volume effects. Analysis using scaled particle theory is consistent with this mechanism. The effects of high concentrations of polymers on the interactions between ribosomal species may make important changes in the function of ribosomes under the crowded conditions which occur in vivo. PMID- 3041373 TI - Identification and sequence of gene dicB: translation of the division inhibitor from an in-phase internal start. AB - The dicA1 mutation, located in the replication termination region of Escherichia coli at 34.9 min, confers a temperature-sensitive, division defective phenotype to its hosts. Previous analysis had suggested that dicA codes for a repressor of a nearby division inhibition gene dicB. We show now that gene dicB is part of a complex operon. Five open reading frames (ORFs 1 to 5) preceeded by a promoter sensitive to dicA repression are found within a 1500 bp segment, and are organized into two clusters separated by a long untranslated region. Evidence for expression of these ORFs was obtained from in vitro or in vivo translation of plasmid-coded genes. IPTG-dependent cell filamentation was obtained when either the entire or the C-terminal part of the fourth ORF was placed under control of the lac promoter. In both cases, a 7 KD protein corresponding to translation from an in-frame ATG of ORF4 (dicB) was made. We propose that this C-terminal protein is the division inhibitor synthesized in dicA1 mutants. PMID- 3041374 TI - Close association of a DNA replication origin and an ARS element on chromosome III of the yeast, Saccharomyces cerevisiae. AB - Two dimensional gel electrophoretic techniques were used to locate all functional DNA replication origins in a 22.5 kb stretch of yeast chromosome III. Only one origin was detected, and that origin is located within several hundred bp of an ARS element. PMID- 3041376 TI - Rapid, large-scale purification and characterization of 'Ada protein' (O6 methylguanine-DNA methyltransferase) of E. coli. AB - The E. coli Ada protein (O6-methylguanine-DNA methyltransferase) has been purified using a high-level expression vector with a yield of about 3 mg per liter of E. coli culture. The 39-kDa protein has an extinction coefficient (E280 nm (1%)) of 5.3. Its isoelectric point of 7.1 is lower than that predicted from the amino acid content. The homogeneous Ada protein is fully active as a methyl acceptor from O6-methylguanine in DNA. Its reaction with O6-methylguanine in a synthetic DNA has a second-order rate constant of 1.1 x 10(9) M-1 min-1 at O degree C. Both the native form and the protein methylated at Cys-69 are monomeric. The CD spectrum suggests a low alpha-helical content and the radius of gyration of 23 A indicates a compact, globular shape. The middle region of the protein is sensitive to a variety of proteases, including an endogenous activity in E. coli, suggesting that the protein is composed of N-terminal and C-terminal domains connected by a hinge region. E. coli B has a higher level of this protease than does K12. PMID- 3041375 TI - Influence of fd gene 2-protein and the viral replication origin on the compatibility of pfd-plasmids. AB - Plasmids with the replication origin of bacteriophage fd, the pfd-plasmids, were investigated for compatibility in E. coli cells expressing fd gene 2-protein. This was measured by transformation of Ca-treated cells with and without a residing pfd-plasmid. When the two plasmids contained the complete intergenic region of bacteriophage fd, they were fully compatible in contrast to the situation in which at least one plasmid had a shortened origin for viral strand replication. This incompatibility effect was partially compensated for by a pfd plasmid with a short origin and with the fd gene 2. The fd replication origin on a colEl plasmid did not affect compatibility in polA+ cells indicating its idling in the presence of the colEl origin. It can be concluded that a short replication origin requires high amounts of gene 2-protein in contrast to the long origin. Accumulation of replication intermediates severely interferes with host cell metabolism. PMID- 3041377 TI - Replication of single-stranded DNA templates by primase-polymerase complexes of the yeast, Saccharomyces cerevisiae. AB - A partially purified primase-polymerase complex from the yeast, Saccharomyces cerevisiae, was capable of replicating a single stranded circular phage DNA into a replicative form with high efficiency. The primase-polymerase complex exhibited primase activity and polymerase activity on singly primed circular ssDNA as well as on gapped DNA. In addition, it was able to replicate an unprimed, single stranded, circular phage DNA through a coupled primase-polymerase action. On Biogel A-O.5m filtration the primase-polymerase activities appeared in the void volume, demonstrating a mass of greater than 500 kilodaltons. Primase and various primase-polymerase complexes synthesized unique primers on single stranded DNA templates and the size distribution of primers was dependent on the structure of the DNA and the nature of the primase-polymerase assembly. PMID- 3041378 TI - The fidelity of base selection by the polymerase subunit of DNA polymerase III holoenzyme. AB - In common with other DNA polymerases, DNA polymerase III holoenzyme of E. coli selects the biologically correct base pair with remarkable accuracy. DNA polymerase III is particularly useful for mechanistic studies because the polymerase and editing activities reside on separate subunits. To investigate the biochemical mechanism for base insertion fidelity, we have used a gel electrophoresis assay to measure kinetic parameters for the incorporation of correct and incorrect nucleotides by the polymerase (alpha) subunit of DNA polymerase III. As judged by this assay, base selection contributes a factor of roughly 10(4)-10(5) to the overall fidelity of genome duplication. The accuracy of base selection is determined mainly by the differential KM of the enzyme for correct vs. incorrect deoxynucleoside triphosphate. The misinsertion of G opposite template A is relatively efficient, comparable to that found for G opposite T. Based on a variety of other work, the G:A pair may require a special correction mechanism, possibly because of a syn-anti pairing approximating Watson Crick geometry. We suggest that precise recognition of the equivalent geometry of the Watson-Crick base pairs may be the most critical feature for base selection. PMID- 3041380 TI - Chimeric multispecific DNA methyltransferases with novel combinations of target recognition. AB - DNA target recognizing domains of different multispecific DNA-cytosine methyltransferases can be rearranged through engineering of the corresponding genes to generate enzymes with novel combinations of target recognition. PMID- 3041379 TI - RepA protein- and oriR-dependent initiation of R1 plasmid replication: identification of a rho-dependent transcription terminator required for cis action of repA protein. AB - Initiation of R1 plasmid replication is dependent on cis-acting repA protein and the 188 base-pair (bp) sequence, oriR. RepA protein synthesized in vitro preferentially activates oriR in cis, regardless of the orientation and location of oriR on the template DNA. RepA protein is not reusable after it activates oriR in the cis-position. Cis-action of repA protein is also dependent on the presence of CIS, a 170 bp sequence, between repA and oriR. CIS contains a rho dependent transcription terminator of the repA transcript, deletion of which results in decrease in transformation efficiency and rapid loss of plasmid in the absence of selection. The significance of transcription termination events in replication was indicated by decreased replication activity in vivo caused by premature termination of the repA transcript between repA and CIS. A model which may account for the role of CIS in mediating the cis-action of the repA protein is presented. PMID- 3041381 TI - The nursing role in radiation oncology: symptom management of acute and chronic reactions. PMID- 3041382 TI - Pregnancy and parenthood after treatment for breast cancer. PMID- 3041384 TI - Buspirone: an update on a unique anxiolytic agent. AB - Buspirone (Buspar) is a azaspirodecanedione anxiolytic agent. Its mechanism of action is extremely complex, but current investigations indicate that its main neuropharmacologic effects are mediated by the 5-HT1A receptors. Other neuroreceptor systems could be involved, as buspirone displays some affinity for DA2 autoreceptors and 5-HT2 receptors. It has been proposed that inhibition of synthesis and release of serotonin result through the combined interactions of neuroreceptors and secondary messenger systems. This action leads to inhibition of the firing rate of 5-HT-containing neurons in the dorsal raphe. From this novel profile, that differs from that of the benzodiazepines, buspirone lacks anticonvulsant and muscle-relaxant properties, and causes only minimal sedation. The drug is rapidly absorbed after oral administration, with a mean bioavailability of 3.9%. After a single oral dose, the mean elimination half-life is 2.1 hours. Buspirone is mainly bound to albumin and alpha 1-acid glycoprotein. It is metabolized to an active metabolite 1-(2-pyrimidinyl) piperazine (1-PP). The mean elimination half-life of 1-PP is 6.1 hours. Buspirone is indicated in the treatment of generalized anxiety disorders. Its efficacy is comparable to the benzodiazepines. Its use in depression and panic disorders requires further investigation. When combined with alcohol or given alone, psychomotor impairment was not detected. Abuse, dependence, and withdrawal symptoms have not been reported. The frequency of adverse effects is low, and the most common effects are headaches, dizziness, nervousness, and lightheadness. Buspirone should be added to drug formularies and could represent a significant addition in psychopharmacology. PMID- 3041383 TI - Nursing care of AIDS patients participating in a Phase I/II trial of recombinant human granulocyte-macrophage colony stimulating factor. PMID- 3041385 TI - DDAVP in the treatment of bleeding disorders. AB - Hemophilia A and von Willebrand's disease are hereditary disorders associated with qualitative and quantitative abnormalities of clotting factor VIII. A major clinical feature is excessive or abnormal bleeding often necessitating the use of transfusions of pooled blood products to achieve hemostasis. Exposure to blood products places the recipient at risk for infection by the hepatitis B virus or the human immunodeficiency virus. A synthetic analog of arginine vasopressin, 1 desamino-8-D-arginine vasopressin, has been shown to increase the plasma levels of factor VIII coagulant activity and von Willebrand's factor, and clinically to improve abnormal bleeding, obviating the need to use blood products. PMID- 3041386 TI - The association between immunodeficiency and congenital heart disease. AB - The predilection of children with congenital heart disease (CHD) to infection may be explained in part by an underlying immunodeficiency disorder. Some 13 syndromes in which immunodeficiency and CHD may coexist have been reported in the medical literature. In addition, immunoglobulin and T-cell deficiencies have been found in nonsyndromal patients with CHD. The diagnosis of immunodeficiency should be entertained in such children, as early recognition of an immunodeficiency disorder can result in improved antimicrobial and immunological management. PMID- 3041387 TI - Cardiovascular drugs in children: angiotensin-converting enzyme inhibitors. AB - Angiotensin-converting enzyme inhibitors are potent vasodilators acting by inhibition of production of the vasoconstrictor angiotensin II. In adults, they are used for treatment of systemic hypertension and congestive heart failure and investigated for treatment of primary pulmonary hypertension. In infants and children, saralasin and captopril were found to be useful in treatment of systemic arterial hypertension, especially when associated with high plasma renin activity. Captopril has failed in the treatment of congestive heart failure associated with complex congenital heart diseases and in most cases of primary pulmonary hypertension. It has a clear beneficial effect in coarctation of the aorta and may have such an effect in endomyocardial diseases and ventricular septal defect. In adults, serious side effects have limited the use of captopril. New converting enzyme inhibitors, devoid of a sulfhydryl group, are expected to have a better safety profile. PMID- 3041388 TI - Immediate pigment-darkening reaction. AB - The immediate pigment-darkening reaction (IPD) is a gray-brown discoloration of the skin induced mainly by long-wave ultraviolet (UVA) radiation. The threshold value of IPD can be used as an important parameter in a series of light tests. The principle mechanism of the reaction is a photo-oxidative process causing darkening of preformed melanin and the production of free radicals. It has also been suggested that the reaction involves micromorphological changes in the melanocytes and a changed distribution pattern of melanosomes in the keratinocytes. This concept has recently been questioned by independent investigators. The physiological function of IPD still remains largely unknown. No photoprotective effect of the reaction has been observed. It has been suggested that IPD might have been of value during human evolution. PMID- 3041389 TI - Transpedicular segmental fixation: description of a new procedure. AB - The treatment of unstable fractures of thoracolumbar vertebrae is a controversial issue in the orthopedic community. The various methods employed for operative stabilization of these injuries have to date been found to have major disadvantages. A new therapeutic principle and a new device for stable internal fixation are presented. PMID- 3041390 TI - The use of cement forms to control methylmethacrylate in immediate posterior stabilization of the cervical spine. A technical note. AB - Under certain clinical conditions, posterior stabilization of the cervical spine, supplemented with polymethylmethacrylate (PMMA), is an accepted method of achieving stability. A technique is described in which "cement forms" are used to limit the spread of PMMA and to allow bone graft placement in the lateral paraspinous gutters. PMID- 3041391 TI - The musculoskeletal manifestations of progeria. A literature review. AB - Although a superficial similarity exists between the musculoskeletal disorders associated with natural aging and those of progeria, an in-depth analysis reveals profound differences in the pathophysiology between the two processes. The protean manifestations of progeria can best be explained on the basis of the vascular changes found at autopsy. A disorder of the vascular endothelium may predispose progeric vessels to atherosclerotic changes. The unique musculoskeletal manifestations of progeria arise from the effects of premature atherosclerosis on the vascularized connective tissues. PMID- 3041392 TI - A rationale for the surgical treatment of bunions. PMID- 3041393 TI - Congenital hemangiopericytoma: report of a case. AB - A congenital hemangiopericytoma of the lower lip, first detected by prenatal ultrasound, was only partially resected and involuted over the next 20 months. PMID- 3041394 TI - Thrombosis of the umbilical cord: analysis of 52 cases and literature review. AB - Fifty-two cases of umbilical cord thrombosis from 3 patients populations are analyzed and compared with 68 cases from the literature. The incidence of cord thrombosis is approximately 1/1300 deliveries, 1/1000 perinatal autopsies, and 1/250 high-risk gestations. There is a slight male predominance. Umbilical vein thrombosis occurs more frequently than thrombosis of one or both umbilical arteries, but poor fetal outcome is more likely with arterial thrombosis. The mechanism of fetal death when only one umbilical artery is thrombosed is illustrated and discussed. The strong association between cord thrombosis and perinatal morbidity and mortality is not noted among prospective cases but, when present, is related to additional umbilical cord abnormalities, obstetrical complications, or systemic fetal conditions that are the likely cause of both the thrombosis and the poor fetal outcome. The pathogenetic relationship between cord thrombosis and these associated conditions is discussed, and it is concluded that cord thrombosis is a marker of both the severity of these conditions and the likelihood of poor fetal outcome. PMID- 3041395 TI - Cardiovascular pathology in osteogenesis imperfecta type IIA with a review of the literature. AB - Lethal perinatal osteogenesis imperfecta (OI Type II) is a biochemically diverse collagen disorder characterized by short, crumpled long bones, beaded ribs, blue sclerae and thin, fragile skin. Cardiovascular abnormalities are rarely described. Using morphometry and light and electron (SEM and TEM) microscopy, we analyzed the hearts and great vessels from 2 fetuses with OI Type IIA and compared the findings with age-matched controls. The heart weights and atrioventricular valve (AVV) circumferences were reduced in OI. The chordae tendineae were short and fragile; both the AVVs and the chordae tendineae were hypercellular. TEM showed relatively little organized collagen in the chordae tendineae of OI fetuses. Furthermore, quantitative evaluation of collagen fibril size revealed a decrease in the cross-sectional diameter. There was also a marked decrease in the adventitial and intramural collagen of the intramyocardial arteries and great vessels in OI. Our study reports, for the first time, specific lesions in the cardiovascular systems of patients with OI Type II and reviews the cardiovascular pathology in other forms of OI. PMID- 3041397 TI - Endoscopic therapy for gastrointestinal bleeding. AB - The use of endoscopic techniques to control gastrointestinal bleeding is an exciting addition to the therapeutic armamentarium. Alternatives include lasers, bipolar probes, heater probes, and sclerosing agents. The lack of statistically significant data should be viewed cautiously, because it is difficult to devise controlled trials for gastrointestinal bleeding that incorporate enough patients to avoid a type II statistical error. However, a healthy degree of skepticism is necessary to prevent the use of highly remunerative techniques of questionable benefit to the patient. PMID- 3041396 TI - Esophageal candidiasis. Managing an increasingly prevalent infection. AB - Esophageal candidiasis is an opportunistic infection that is being recognized increasingly often in certain patients, including those who have a neoplastic disease, are undergoing protracted antibiotic therapy, or hae acquired immunodeficiency syndrome (AIDS). Impaired cell-mediated immunity may predispose the patient to esophageal mucosal colonization, whereas chemotherapy-induced granulocytopenia may predispose to disseminated candidiasis. Esophageal candidiasis should be suspected in susceptible patients with complaints of substernal odynophagia or dysphagia. The diagnosis is confirmed by endoscopically directed mucosal biopsy. Esophagitis from other causes (eg. herpes simplex virus, cytomegalovirus, or bacterial infection) may develop concomitantly with esophageal candidiasis. Treatment is determined by the clinical and immune status of the patient. Amphotericin B (Fungizone) is administered to immunocompromised patients at risk for disseminated or deeply invasive candidiasis and is indicated in nongranulocytopenic patients whose symptoms prevent reliable administration of oral antifungal agents. Ketoconazole (Nizoral) may be administered to clinically stable nongranulocytopenic patients with esophageal candidiasis limited to the mucosa. Patients with AIDS and a history of esophageal candidiasis usually benefit from long-term suppression with an oral antifungal agent. PMID- 3041398 TI - Interactions between calcium channel blockers and non-cardiovascular drugs: interactions with anticancer drugs. PMID- 3041399 TI - Cholecystokinin (CCK)-33 stimulates insulin secretion from the perfused rat pancreas: studies on the structure-activity relationship. AB - Cholecystokinin (CCK)-33 is known to stimulate insulin secretion. Presently, using the perfused rat pancreas, we have characterized the active site in the CCK 33 molecule that is responsible for this effect by the use of different CCK fragments. We found that CCK-33, CCK-8 and CCK-7 (1 nM) all significantly stimulated insulin secretion in the presence of 4.4 mM or 6.7 mM glucose. However, CCK-7 was much less potent than the longer forms. In contrast, CCK-4, CCK-6 and CCK-33 (1-21) had no effect on insulin secretion. We conclude that the shortest CCK-form that stimulates insulin secretion at 1 nM is the C-terminal heptapeptide CCK-7. However, CCK-8 is much more potent than CCK-7 in this respect. PMID- 3041400 TI - [Guidelines on diagnostic bronchoalveolar lavage]. PMID- 3041401 TI - [Platelet activating factor and its possible role in bronchial asthma]. PMID- 3041402 TI - Single umbilical artery: prenatal findings. AB - In 450 patients with pregnancy at high risk for fetal malformation and/or intrauterine growth retardation, the umbilical cord was investigated sonographically for the presence of a single umbilical artery. A single umbilical artery was diagnosed in four fetuses between 23 and 33 weeks of gestation and suspected in two. Three cases were overlooked at sonography. All seven surviving fetuses had growth retardation at delivery and four also showed severe malformations. Whenever a single umbilical artery is found at sonography, further work-up is required to rule out associated anomalies, intrauterine growth retardation, or chromosomal abnormality. PMID- 3041403 TI - Immunoblot detection of enzyme proteins of peroxisomal beta-oxidation in fibroblasts, amniocytes, and chorionic villous cells. Possible marker for prenatal diagnosis of Zellweger's syndrome. AB - The amounts of enzyme proteins of peroxisomal beta-oxidation in fibroblasts and chorionic villous cells from infants with Zellweger syndrome and in fibroblasts, amniocytes, and chorionic villi from healthy controls were measured by immunoblot analysis. Immunoreactive proteins of peroxisomal acyl-CoA oxidase and 3-ketoacyl CoA thiolase were absent in fibroblasts and chorionic villous cells from the patients, yet these enzyme proteins were present in fibroblasts, cultured amniocytes, and chorionic villi from the normal controls. These results show that immunoblot analysis of peroxisomal beta-oxidation enzymes in amniocytes and chorionic villous cells is of potential value for the prenatal detection of Zellweger syndrome. PMID- 3041404 TI - Spontaneous resolution of a cystic hygroma in a fetus with Turner syndrome. AB - The prenatal detection of a cystic hygroma (CH) in a fetus with a 45,X karyotype is described. The cystic hygroma underwent spontaneous resolution and a healthy baby with Turner syndrome was subsequently born. The implications for genetic counselling are discussed. PMID- 3041406 TI - Regulatory aspects of placental iron transfer--a comparative study. AB - Receptor-mediated endocytosis is generally assumed to be the process by which the haemochorial placenta takes up iron from transferrin. The involvement of an additional nonendocytic process cannot, however, be excluded. It appears from a study of iron transport mechanisms and of the maturation of the transfer process that placental ferritin is involved in the transfer of iron from mother to fetus. The metabolic relationship between the ferritin pool and the placental transfer pool remains to be elucidated. There is no evidence for short-term regulation of placental transfer capacity in response to changes in the maternal iron supply or to changes in the trophoblastic iron content. This cannot yet be said of fetal feedback control of placental iron uptake because the experiments performed so far do not permit conclusions on this point. The capacity for iron uptake and transfer seems to increase in accordance with the ontogenetically determined placental growth pattern. This does not exclude long-range adaptive modifications of the transfer capacity in response to early maternal or fetal disturbances. The results obtained from studying placental maturation suggest that a possible long term regulatory interaction between growing placenta and fetus may occur. Clinical evidence so far is inconclusive. The relatively moderate reductions in the fetal iron stores which are generally associated with severe iron-deficient pregnancies might be seen as an argument in favour of long-term placental control. The marked impact of pregnancy on maternal iron metabolism in rodents, as compared to other mammals, is possibly met by means of direct placental control of mucosal iron uptake. In primates, mucosal iron uptake during pregnancy seems to be governed by factors related to systemic iron deficiency only. PMID- 3041405 TI - Immunohistochemical studies of fetal trophoblast and maternal decidua in hydatidiform mole and choriocarcinoma. AB - Immunohistochemical techniques have been used to investigate the expression of fetal trophoblast antigens and the maternal leucocytic response in molar pregnancy and choriocarcinoma. The antigenic phenotype of morphologically defined trophoblast populations in complete, partial and invasive moles was analogous to that in normal pregnancy. All trophoblast phenotypes described in normal pregnancy were also identified in choriocarcinoma, suggesting that extensive differentiation into heterogeneous subgroups occurs in malignant trophoblast. The maternal leucocytic infiltrate in molar pregnancy consisted of T lymphocytes and class II MHC-positive macrophages. CD2-positive, CD3-negative lymphocytes were identified in molar decidua but not in uterine tissue in choriocarcinoma. Similarly, endometrial granulocytes were present in molar decidua but not in choriocarcinoma; these cells were associated with decidualization rather than with fetal trophoblast. PMID- 3041407 TI - Site-directed mutations altering methyl-accepting residues of a sensory transducer protein. AB - The Trg protein is one of a family of transducer proteins that mediate chemotactic response in Escherichia coli. Transducers are methyl-accepting proteins that gain or lose methyl esters on specific glutamyl residues during sensory adaptation. In this study, the significance of multiple sites of methylation on transducer proteins was addressed by using oligonucleotide directed, site-specific mutagenesis to substitute an alanyl residue at each of the five methyl-accepting sites in Trg. The resulting collection of five mutations, each inactivating a single site, was analyzed for effects on covalent modification at the remaining sites on Trg and for the ability of the altered proteins to mediate sensory adaptation. Most of the alanyl substitutions had substantial biochemical effects, enhancing or reducing methyl-accepting activity of other sites, including one case of activation of a site not methylated in wild type protein. Analysis of the altered proteins provided explanations for many features of the complex pattern of electrophoretic forms exhibited by Trg. The mutant proteins were less efficient than normal Trg in mediating adaptation. Correlation of biochemical and behavioral data indicated that reduction in the number of methyl-accepting sites on the transducer lengthened the time required to reach an adapted state. PMID- 3041408 TI - Discrete-time random walks on diagrams (graphs) with cycles. AB - After a review of the diagram method for continuous-time random walks on graphs with cycles, the method is extended to discrete-time random walks. The basic theorems carry over formally from continuous time to discrete time. Three problems in tennis probabilities are used to illustrate random walks on discrete time diagrams with cycles. PMID- 3041409 TI - In vivo "photofootprint" changes at sequences between the yeast GAL1 upstream activating sequence and "TATA" element require activated GAL4 protein but not a functional TATA element. AB - Transcription of the yeast GAL1 and GAL10 genes is induced by growth on galactose. Using the technique of photofootprinting in vivo, we previously documented equivalent transcription-dependent footprints within the putative "TATA" elements of both genes. To explore the functional significance of these observations, we created a 3-base-pair substitution mutation within the GAL1 promoter TATA element, which disrupted the ATATAA consensus sequence but left intact the photomodification targets. The mutation reduced galactose-induced RNA levels by a factor of 100. The mutant promoter no longer displayed the characteristic TATA sequence footprint, supporting the hypothesis that transcription activation involves the binding of a TATA box factor. We also observed a collection of transcription-correlated alterations in the modification pattern at sites between the UASG and the GAL1 TATA element, within sequences that are not required for inducible transcription. These patterns, characteristic of the induced wild-type GAL1 gene, were still galactose inducible with the TATA mutant GAl1 promoter, despite the low level of transcription from this promoter. We conclude that the GAL4-dependent protein/DNA structure responsible for the altered pattern within nonessential sequences is therefore not strictly coupled to an active TATA element or to high levels of expression. Nonetheless, the patterns probably reflect a stable protein-dependent structure that accompanies assembly of the transcription initiation complex. PMID- 3041410 TI - Alternative DNA loops regulate the arabinose operon in Escherichia coli. AB - The araCBAD regulatory region of Escherichia coli contains two divergently oriented promoters and three sites to which AraC, the regulatory protein of the operon, can bind. This paper presents the results of in vivo dimethyl sulfate "footprinting" experiments to monitor occupancy of the three AraC sites and measurements of activity of the two promoters. These measurements were made both in the absence of the inducer arabinose and at various times after arabinose addition to growing cells containing the wild-type ara regulatory region or the regulatory region containing various deletions and point mutations. The data lead to the conclusion that two different DNA loops can form in the ara regulatory region. These loops are generated by AraC protein molecules binding to two different DNA sites and binding to each other. One of these loops predominates in the absence of arabinose and plays a major role in repressing activity of one of the promoters. Upon the addition of arabinose the amount of the first loop type, the repression loop, decreases and the amount of a second loop increases. Formation of this second loop precludes the counterproductive formation of the repression loop. PMID- 3041411 TI - Relationship between endo- and exopeptidases in a processing enzyme system: activation of an endoprotease by the aminopeptidase B-like activity in somatostatin-28 convertase. AB - The somatostatin-28 convertase activity involved in vitro in the processing of somatostatin-28 into the neuropeptides somatostatin-28-(1-12) and somatostatin-14 is composed of an endoprotease and a basic aminopeptidase. We report herein on the purification to apparent homogeneity of these two constituents and on their functional interrelationship. In particular we observed that after various physicochemical treatments, the 90-kDa endoprotease activity was recovered both at this molecular mass and as a 45-kDa entity. Moreover, the production of [Arg 2,Lys-1]somatostatin-14 from somatostatin-28 by the action of the endoprotease was activated in a cooperative manner by the aminopeptidase B-like enzyme. A 10 fold activation occurred when the exopeptidase was inhibited by 6.5 mM diisopropyl fluorophosphate and allowed the determination of a half-maximal activation constant (K1/2) of approximately equal to 13 nM. These observations strongly suggest that both enzymes act in a concerted manner in vitro and that they may form a complex in vivo. PMID- 3041412 TI - Crosstalk between bacterial chemotaxis signal transduction proteins and regulators of transcription of the Ntr regulon: evidence that nitrogen assimilation and chemotaxis are controlled by a common phosphotransfer mechanism. AB - We demonstrate by using purified bacterial components that the protein kinases that regulate chemotaxis and transcription of nitrogen-regulated genes, CheA and NRII, respectively, have cross-specificities: CheA can phosphorylate the Ntr transcription factor NRI and thereby activate transcription from the nitrogen regulated glnA promoter, and NRII can phosphorylate CheY. In addition, we find that a high intracellular concentration of a highly active mutant form of NRII can suppress the smooth-swimming phenotype of a cheA mutant. These results argue strongly that sensory transduction in the Ntr and Che systems involves a common protein phosphotransfer mechanism. PMID- 3041413 TI - Antibody to sigma 32 cross-reacts with DnaK: association of DnaK protein with Escherichia coli RNA polymerase. AB - A polyclonal antibody to sigma 32, the heat shock sigma factor, has been used to show the presence of low levels of sigma 32 in Escherichia coli RNA polymerase preparations (E sigma 70), which explains the observed in vitro activity of E sigma 70 towards heat shock genes. The sigma 32 antibody cross-reacts with DnaK, and DnaK has been found associated with purified preparations of both E sigma 70 and the heat shock RNA polymerase, E sigma 32. PMID- 3041414 TI - Inhibition of human immunodeficiency virus replication by antisense oligodeoxynucleotides. AB - Twenty different target sites within human immunodeficiency virus (HIV) RNA were selected for studies of inhibition of HIV replication by antisense oligonucleotides. Target sites were selected based on their potential capacity to block recognition functions during viral replication. Antisense oligomers complementary to sites within or near the sequence repeated at the ends of retrovirus RNA (R region) and to certain splice sites were most effective. The effect of antisense oligomer length on inhibiting virus replication was also investigated, and preliminary toxicity studies in mice show that these compounds are toxic only at high levels. The results indicate potential usefulness for these oligomers in the treatment of patients with acquired immunodeficiency syndrome (AIDS) and AIDS-related complex either alone or in combination with other drugs. PMID- 3041416 TI - Expression of streptococcal M protein in mammalian cells. AB - The M protein encoded by group A streptococci is a cell-wall polypeptide that has the property of enabling these organisms to evade the phagocytic cells of the human host. Therefore, the M protein plays a major role in the pathogenesis of streptococcal diseases. As an initial step toward the use of this protein as a target antigen for the production of protective anti-streptococcal immunity, a live vaccinia virus recombinant containing the M-protein gene has been constructed (VV:M6 delta). The bacterial M-protein DNA sequence is stable within this genetic context and is actively transcribed by viral RNA polymerase. Furthermore, high levels of immunoreactive M protein were detected in vivo when the VV:M6 delta recombinant was used to infect mammalian cells in culture. Thus, in addition to providing a powerful approach for dissecting the immunodominant domains of the M protein, the VV:M6 delta recombinant appears to be an excellent candidate vaccine for animal trials. PMID- 3041417 TI - [Active oxygen species. Significance for disease and health]. PMID- 3041415 TI - Cinnamyl-alcohol dehydrogenase, a molecular marker specific for lignin synthesis: cDNA cloning and mRNA induction by fungal elicitor. AB - Cinnamyl-alcohol dehydrogenase (CAD; EC 1.1.1.195) catalyzes the final step in a branch of phenylpropanoid synthesis specific for production of lignin monomers. We have isolated a full-length cDNA clone encoding CAD, as a molecular marker specific for lignification, by immunoscreening a lambda gt11 library containing cDNAs complementary to mRNA from elicitor-treated cell cultures of bean (Phaseolus vulgaris L.). The clone comprises a single long open reading frame of 1767 base pairs, 31 base pairs of 5' leader, and 152 base pairs of 3' untranslated sequence. The deduced 65-kDa CAD polypeptide has several features that are strongly conserved in alcohol dehydrogenases. Addition of fungal elicitor to cell cultures stimulates CAD transcription, which leads to a remarkably rapid, but transient, accumulation of CAD mRNA, with no detectable lag and maximal levels after 1.5 hr. Southern blot analysis of bean genomic DNA indicates that elicitor-induced CAD is encoded by a single gene. The regulatory significance of the rapid activation of this CAD gene and the possible existence of a second, divergent CAD gene involved in lignification during xylogenesis are discussed. PMID- 3041419 TI - [The genesis of clinical nursing in Poland in the period between the wars]. PMID- 3041418 TI - Influence of hyperthermia, pH and culturing conditions on the electrical parameters of Chinese hamster V79 cells. AB - Conductivity measurements in the frequency range 10 kHz-100 MHz have been performed on Chinese hamster fibroblasts V79 exposed to hyperthermia in different experimental conditions. The membrane electrical conductivity is decreased when cells are heated for 1 h at 44 degrees C or 3 h at 43 degrees C at pH 7.25. This effect has not been observed on cell samples following heating for 1 h at 43 degrees C at pH 7.25. However, if the pH of the culture medium is adjusted to 6.50 or 6.0 before hyperthermic exposure, a substantial decrease of membrane conductivity is observed. In cells maintained in partially spent medium before heating, a 3 h treatment at 43 degrees C at pH 7.25 does not produce any effect on membrane conductivity. The results of all these experiments seem to indicate that some alteration of the energy-driven mechanisms involved in active transport across the plasma membranes may be responsible for the observed effects. PMID- 3041420 TI - [Halina Antonowicz]. PMID- 3041421 TI - [Price list for midwives from 160 years ago]. PMID- 3041422 TI - [Outline of the history of the midwife profession to the 20th century (I)]. PMID- 3041423 TI - [The first midwife regulations from 1827]. PMID- 3041424 TI - [40th anniversary of the Mieczyslaw Michalowicz Medical School No. 1]. PMID- 3041425 TI - [Outline of the history of the midwife profession to the 20th century]. PMID- 3041426 TI - [The requirements set for the midwife candidate 170 years ago]. PMID- 3041427 TI - Maqua (therapeutic burn) as an indicator of underlying disease. AB - The origin and nature of the maqua (the Arabic therapeutic burn) is presented together with our clinical experience of patients previously treated by this traditional method. Maquas are small deep burns inflicted in areas either in proximity to a diseased organ or in points related traditionally to the original basic problem. These relationships may be rooted in historical ties between old Arab medicine and traditional Oriental, antique Egyptian, and Greco-Roman medicines. Maquas alone only rarely present a threat to the patient, but in many cases they may serve as an indicator of the original underlying disease. This and other folklore treatment modalities, together with the healers themselves, should be acknowledged by us, as markers for health problems or maybe for potential healing methods and doctor-patient relationships. PMID- 3041429 TI - A simple, rapid technique for skin grafting using an adhesive transparent dressing. PMID- 3041430 TI - Primary care as the middle ground for psychiatric epidemiology. PMID- 3041428 TI - The antifibrinolytic activity of sulfamylon solution. AB - Sulfamylon (mafenide) solution, a potent experimental topical antimicrobial, is used in our burn unit to treat burn wounds both before and after skin grafting. The importance of fibrin to early graft adherence prompted this in vitro study of the effect of Sulfamylon upon fibrin clot. Assessing fibrinolysis by in vitro proteolysis of [125I] fibrin monomers, Sulfamylon, at relevant clinical concentrations, produced dose-related inhibition of streptokinase-mediated fibrinolysis. In addition, Sulfamylon had no intrinsic fibrinolytic activity. This antifibrinolytic property of Sulfamylon solution may, in vivo, protect against early graft loss when used on burn wounds and other potentially contaminated graft sites. PMID- 3041432 TI - Schizophrenia before 1800? The case of the Revd George Trosse. AB - Trosse's account of his life has been described as one of the strangest pieces of realism in the English language. It gives a detailed description of the author's personal experience of three episodes of psychotic disorder during the years 1656 7. The disorder has been considered an early instance of schizophrenia, but evidence is presented here to suggest that a more plausible diagnosis was alcoholic psychosis and affective disorder. PMID- 3041431 TI - Expressed emotion: current status. PMID- 3041433 TI - Inter-task consistency: an integrative re-evaluation. PMID- 3041434 TI - The G. W. Harris lecture. Steroid control of brain and pituitary function. PMID- 3041435 TI - [Dependence of the radiosensitivity of yeast cells on the LET of radiations. Experiments on haploid cells]. AB - The previously developed model was used to study the dependence of radiosensitivity (D0(-1) of Saccharomyces cerevisiae (the wild type and radiosensitive mutant) on linear energy transfer (LET) of ionizing radiation. D0( 1) (L) of haploid yeasts was shown to be associated, to a certain extent, with the capacity of radiation damages repair. As to the wild-type cells, the above function was represented by a curve showing a maximum, while a descending curve was characteristic of the radiosensitive mutant cells deficient in radiation damages repair. The influence of the repair processes on cell radiosensitivity decreased with increasing LET. PMID- 3041436 TI - [Regeneration of the sciatic nerve in the progeny of female rabbits irradiated prior to mating]. AB - A study was made of the sciatic nerve regeneration after smooth cutting thereof in 198 young chinchillas whose mothers were subjected to whole-body single irradiation with doses of 0.05, 0.15, 0.5 or 1.5 Gy one week before mating. The statistically significant data obtained may be used in estimating a risk from ionizing radiation and ensuring the radiation security standards. PMID- 3041438 TI - [Cytogenetic research on micropopulations of the root vole (Microtus oeconomus Pall.) living under various radioecological habitat conditions]. AB - Percentage of aneuploid cells and that of cell with chromosome aberrations in Microtus oeconomus Pall. living in areas with the enhanced radiation background (the dose of external radiation is 50 times and that of internal irradiation of bone marrow by incorporated 226Ra 10 times higher than the controls) exceeds significantly (p less than 0.05) the control values. It is concluded that mutations occur in the experimental animals more frequently than in the controls. PMID- 3041437 TI - [Late sequelae of exposure to ionizing radiation with various LET's. Effect of dose rate and fractionation on changes in the life span of rats]. AB - In experiments with 2120 albino mongrel rats their life span was followed up after the effect of various types of radiation (for instance, gamma-neutron radiation of 0.9 MeV and gamma- and X-rays) at different exposure schedules (that is, whole-body irradiation with doses from LD0/30 to LD100/30 and fractionated at 24 and 72 hour intervals and dose--rates varying from 0.00042 Gy/min to 1.02 Gy/min). The type of radiation, the dose--rate, single and cumulative doses, the number of fractions and the interval between them were estimated with respect to their contribution to life span shortening. PMID- 3041440 TI - [Postradiation life span of mice in relation to the modifying action of leukemic cells administered]. AB - Leukemic cells administered to X-irradiated mice modify the radiobiological effect to a degree that depends on the radiation dose, bone marrow transplantation, the quantity and quality of leukemic cells, and the time of their administration. PMID- 3041439 TI - [Effect of millimeter-range radiation on the effectiveness of bone marrow transplantation]. AB - The results presented in this report show that while bone marrow of intact donors transplanted to irradiated (10 Gy) recipient mice produces a 2.5-fold increase in the life span of animals, bone marrow of donors exposed to EHF increases it by 35 times. PMID- 3041441 TI - [Action of alpha-tocopherol on the enzymes of the electron transport chains in the liver microsomes of irradiated rats]. AB - Five days following single whole-body gamma irradiation of rats (8.5 Gy) the rate of NADPH and NADH oxidation, the activity of NADPH-cytochrome P-450 and NADH cytochrome b5 reductases, and the content of cytochromes P-450 and b5 were found to decrease. The intragastric administration of alpha-tocopherol (100 mg/kg, two times a day) produced a normalizing effect. PMID- 3041442 TI - [Functional scintigraphy of the esophagus: value in preoperative diagnosis]. PMID- 3041443 TI - [Cerebral radionuclide angiography--standard procedure and automatic evaluation]. PMID- 3041445 TI - [Parametric imaging in the evaluation of regional cerebral blood circulation]. PMID- 3041444 TI - [Diagnosis of carotid stenosis using radionuclide angiography and ultrasonic tomography]. PMID- 3041446 TI - [Changes in kidney transplant perfusion at the onset of a rejection crisis]. PMID- 3041447 TI - [Fine-needle biopsy of the kidneys]. AB - The methods, indications, results, and complications in fine-needle puncture of the kidneys are discussed, with particular attention to cysts that have an atypical appearance in tomographic procedures. The problems of percutaneous cyst treatment and the possibilities of abscess therapy by fine-needle puncture are presented. The limited indications of using this method in solid kidneys tumors are defined, and antegrade pyelography and the differentiation of perirenal processes are discussed. PMID- 3041448 TI - [Sonography of pancreatic carcinoma. Sonomorphology, diagnostic accuracy and tumor staging]. AB - Ultrasonography was performed in 125 consecutive patients with clinically suspected carcinoma of the pancreas. The most important diagnostic criteria were reflectivity and echo amplitude, which were locally decreased in 71% (50/70) and increased in 27% (19/70) of the cancers. The echogenicity and reflectivity did not differ in four endocrine active tumors and adenocarcinomas. The diagnostic accuracy was 83% (101/122); in 3 patients ultrasonography failed to provide adequate information. Dilation of the biliary tree was found in 68% (43/50) of the carcinomas and dilation of the Wirsungian duct in 52% (32/58). There was some difficulty in differentiating between pancreatic carcinoma and chronic pancreatitis because the echogenicity is similar for both and inflammatory and neoplastic pancreatic processes are also present in both diseases. Although only 2-3 cm in diameter, 87% (7/8) of the carcinomas were already in the T3 stage. Angiography to evaluate resectability is unnecessary only when vascular encasement, extensive infiltration of peripancreatic fat, or tumor thrombus has been demonstrated sonographically. PMID- 3041449 TI - Food-associated intoxicants. AB - The association of toxic substances with human foods has long been recognized. While intrinsic compounds appear during storage as a result of spoilage by chemical processes or by contamination with micro-organisms. In the numerous stages of food production from source to table there are many opportunities for contamination. This article reviews the wide spectrum of food-associated toxicants, outlining the mechanisms by which these substances reach the food products. To illustrate the diversity of these mechanisms, some notable examples of mass contamination of food are quoted. The presence of toxic substances in human food is, and will continue to be, a challenge for toxicologists, and a source of concern for the public, for industry, and for the scientific community. PMID- 3041450 TI - Sensitivity to overfeeding: the Quebec experiment with identical twins. AB - The role of the genotype in the response to short-term overfeeding was assessed by submitted six pairs of male monozygotic twins to a 4.2 MJ (1000 kcal) per day energy intake surplus for a period of 22 consecutive days. Individual differences in fat mass and fat-free mass gains were observed in response to overfeeding but they were not randomly distributed. Indeed, the within-pair resemblance in the response was striking when compared to the heterogeneity found among the pairs in adiposity and fat-free mass gains. The intrapair resemblance in the response to overfeeding as assessed by the intraclass coefficient computed with the individual changes, reached 0.88 for total fat mass and 0.76 for fat-free mass. A similar trend for a genetically determined pattern of adaptation to overfeeding was observed for resting metabolic rate (intraclass = 0.63), thermic effect of a meal (intraclass = 0.62), and energy cost of submaximal exercise (intraclass = 0.78) when the data were analysed in terms of changes in oxygen uptake. On the other hand, no major alterations in glucose and insulin response to a glucose load or a test meal, in cardio-pulmonary adaptation to submaximal exercise and in maximal exercise tolerance were found with overfeeding. In contrast, the response of suprailiac fat cell lipolysis (intraclass of about 0.7) and heparin releasable adipose tissue lipoprotein lipase (intraclass - 0.82) varied among individuals but was highly homogeneous within genotypes. Similarly, a genotype-overfeeding interaction effect was seen for serum triglycerides (intraclass = 0.69), HDL cholesterol (intraclass = 0.85), and the HDL-cholesterol to total cholesterol ratio (intraclass = 0.82). Multiple correlation analyses suggest that much of the variance in the response of fat mass (R = 0.65) and fat-free mass (R = 0.81) is accounted for by alterations in the energy expenditure components assessed in the study. If one takes into account the measurement errors always present in such complex studies and the fact that only a limited fraction of the energy expenditure of activity was considered by design, one can conclude that the genotype determines to a large extent the response variation to short-term overfeeding. In particular, the genotype-overfeeding interaction effect for body composition changes seems to be mediated by the various energy expenditure components, themselves characterized by significant genotype-overfeeding interaction effects.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3041452 TI - Brain development in dwarf mice. PMID- 3041451 TI - Relationship between carbohydrate sweeteners and oral diseases. AB - Dental caries and periodontal disease are wide-spread oral illnesses whose etiology is intimately associated with the consumption of carbohydrate sweeteners. Since dental caries is multifactorial in nature, it is reasonable to assume that there is no single method, usable in the foreseeable future, which would lead to its eradication. Consequently, any new caries preventive measures must function coherently with those currently in use, so that the concerted action of all methods would lead to the best cariostatic effect. Strict restriction of sucrose intake without suggestion of alternatives is not realistic. This fact has given rise to attempts to replace sucrose, especially in products consumed between meals, with sweeteners that are less cariogenic. Human clinical trials and several animal experiments have shown promising clinical results obtained by replacing sucrose with certain sugar alcohols (polyols). Among the sugar alcohols, the best results so far have been obtained with xylitol, which is chemically a pentitol containing five carbon atoms. Chewing gums containing xylitol have been shown to be strong instruments against caries in caries-active age-groups and in high-risk subjects. More research is needed to assess the ability of mixtures of xylitol with sorbitol, palatinit, maltitol, other sugar alcohols, and intense sweeteners to prevent oral plaque diseases. Although thorough clinical trials on the relationship between carbohydrate sweeteners and periodontal diseases have not been performed, the available data indicate that dietary polyols may have a restricted dampening effect on periodontal and gingival inflammations. PMID- 3041453 TI - Purinergic mechanisms in epilepsy. PMID- 3041454 TI - [Introduction to biological calorimetry]. PMID- 3041455 TI - [Time-resolved calorimetry of intermediate steps of enzyme reactions]. PMID- 3041456 TI - [Calorimetry of the ligand binding to proteins]. PMID- 3041458 TI - [Advances in thermodynamic analysis of heat capacity function]. PMID- 3041457 TI - [Thermally induced transitions of biopolymers]. PMID- 3041459 TI - [Heat production of the cell as a parameter for cell function]. PMID- 3041460 TI - [Allergy and its regulation]. PMID- 3041461 TI - [Mechanism of drug resistance in cancer and circumvention of resistance]. PMID- 3041462 TI - [The psychology of femininity. A critical review]. PMID- 3041463 TI - [From the archives of psychoanalysis. By Lillian Rotter. On the psychology of female sexuality (1932)]. PMID- 3041464 TI - [Difficulties in the philosophical reception of Freud]. PMID- 3041465 TI - [Edgar Allan Poe: detection and poet (with reference to Marie Bonaparte)]. PMID- 3041466 TI - [Freud: the nature of man and the social aspects of nature]. PMID- 3041467 TI - Electroconvulsive therapy: a clinical discussion. PMID- 3041468 TI - [Chromatin-level analysis of regulation of gene expression]. PMID- 3041469 TI - [Thrombosis of the inferior vena cava in Behcet's disease. Ultrasonic study]. PMID- 3041470 TI - [Polycystic liver. Clinico-radiological considerations on nine cases in the same family]. PMID- 3041471 TI - [Atypical echographic image of hepatic miliary tuberculosis]. PMID- 3041472 TI - [Jaundice caused by lithiasis in infants. Diagnosis and ultrasonic monitoring]. PMID- 3041473 TI - [Cystic dilatation of the common bile duct. Difficult preoperative diagnosis. Apropos of a case]. PMID- 3041474 TI - [Torsion of the epiploon. Description of a case]. PMID- 3041475 TI - [Alessandro Vallebona (1899-1987)]. PMID- 3041476 TI - [Diagnostic imaging and interventional radiology in abdominal abscess formations]. AB - Abdominal abscesses as a complication of laparotomic surgery have a high mortality rate. The authors reviewed the diagnostic and therapeutic procedures of 36 patients who developed intra-abdominal abscesses after surgical treatment for abdominal neoplasias. The first-step diagnostic procedures (plain film of the abdomen and chest, CT and US) showed a sensibility of 78%. In 25/36 patients (69.5%) two interventional radiology procedures were performed: fine needle aspiration and catheter drainage of the abscess. In 16% of patients fine needle aspiration led to a complete evacuation of the abscess cavity and guaranteed the recovery. In 84% of cases a drainage catheter was positioned into the cavity and left indwelling. This case review is aimed at stressing how plain film of the abdomen is still a diagnostic procedure with high sensibility and specificity for this pathology, even though it is currently considered as a second-choice diagnostic step--US and CT being assessed as the methodologies of choice. The latter techniques can both provide a more accurate imaging when interventional radiology procedures are to be performed. PMID- 3041477 TI - [Ultrasonic-guided fine-needle biopsy of osteolytic lesions]. AB - Percutaneous biopsy of lytic lesions of the bone, in the past pertaining to orthopedic surgeons, has now become a part of interventional radiology. Fluoroscopic guidance has simplified its execution. US does not, under normal conditions, allow an accurate examination of the skeleton; on the other hand, when the bone tissue is replaced by soft tissue, US can easily demonstrate the presence of a tumefaction, caused by a lytic lesion, and determine its characteristics. Eleven patients with lytic lesions (recognized on plain film) underwent US. The lesions were demonstrated. In such cases, biopsy is essential to define their benign or malignant nature, in the latter case if primitive or metastatic, and their histology as well. US is a simple guide to the biopsy of noncorticalized lytic lesions, and it offers consistent advantages over fluoroscopic guidance--i.e. it is easier to perform, it requires no X-ray exposure, it provides with three-dimensional images, the guide is performed in real time, and a choice of the most appropriate (non-necrotic) areas is possible. PMID- 3041478 TI - [Imaging of enthesopathies of the ligamentum patellae in athletes. Echography and computerized telethermography]. AB - The authors examine the diagnostic value of two non-invasive techniques--i.e. ultrasound and computerized teletermography--in the diagnosis of patellar enthesopathies. Such a pathology is quite frequent in young athletes, at various stages. The patients were grouped in two categories, according to their age when the trauma occurred, and to the peculiarities of both enthesopathic localization and imaging. The use of both techniques, either one prevailing over the other according to the different evolutive phases of enthesopathies, provides useful information as to detecting and determining the disease. Finally, the authors recommend a combined use of these techniques, especially in young athletes, possibly avoiding conventional X-rays. PMID- 3041479 TI - [Digital subtraction radiology in the study of mobile structures of the larynx]. AB - Digital subtraction radiography (DSR) was applied to the study of the larynx in 11 healthy subjects and 15 pathological cases. The method, consisting in the subtraction of images obtained at rest and during phonation or respiratory phases, allowed a clear definition of the normal moving structures--i.e. vocal cords, false cords, pyriform sinuses, thyroid cartilage. Moreover, several pathological conditions could be demonstrated. DSR asserts thus itself as a suitable technique in the functional evaluation of the glottis. PMID- 3041480 TI - [Echography and computerized tomography in the diagnosis of complex abdominal lesions]. AB - Complex abdominal lesions include a variety of pathologies, such as septated, infected, and hemorrhagic cysts, abscesses, tumors, and fluid collections of different etiology. These lesions present diagnostic difficulties with both Ultrasonography (US) and Computed Tomography (CT), since findings may not be present or, when present, are not specific. Keeping these limitations in mind, we evaluated 105 patients (111 abdominal lesions) with both US and CT in order to compare their adequacy in predicting the nature of the lesion. On the basis of US and CT results, complex abdominal lesions were divided in four classes: class I includes 43 cases in which both examinations gave the same contribution to the definition of the nature of the lesion, class II (14 lesions, mainly sepimentated cysts), in which US was superior to CT, class III includes 45 cases in which CT was superior to US, mainly in case of hemorrhagic cysts, abscesses, fluid collections and, less frequently, cystic tumors; class IV includes 9 cases in which US and CT results were complementary, which allowed the nature of the lesion to be defined. In conclusion, US and CT enable the identification and the characterization of complex abdominal lesions; an association of the two investigations enhances their diagnostic value. As a rule, CT is superior when the content of the lesion is either gas or hemorrhage, and in the definition of its peripheral wall. US is always superior in assessing septa, and sometimes even vegetations. PMID- 3041482 TI - [Echography in renal colic]. AB - To determine the role of sonography (US) in patients with renal colic, 40 patients were examined by means of US, plain abdominal film (PF), and intravenous pyelography (ivp). US sensitivity was 92.3% in diagnosing hydronephrosis and 75% in detecting calculi. Small calculi were correctly identified, irrespective of their chemical composition. It must be stressed how US, as compared to ivp, proved unsatisfactory in such cases as difficult visualization of the middle portion of the ureter, unsuccessful identification of acute obstructions without hydronephrosis (although the patient's hydratation may be useful in this respect), poor functional information (although there was a correlation between renal hyper-echogenicity and obstructive nephrogram). US is safe and easy to perform, and is suggested for the initial evaluation of patients with renal colic, together with PF, and as an alternative to ivp. Moreover, US is the ideal technique in the follow-up of these patients. Therefore, ivp should be performed in case of differing clinical and sonographic findings, when the calculus is not ejected within the expected time, and when surgery or lithotripsy are foreseen. PMID- 3041481 TI - [Importance of biological findings in the ultrasonic diagnosis of neoplastic biliary obstructions]. AB - The authors' purpose is to demonstrate the possibility of improving US reliability in the diagnosis of neoplastic obstructions of the bile ducts, basing their study on the hematic alkaline phosphatase level (AP), which is an earlier sign of obstruction than high bilirubin values. All 368 patients observed had AP levels above the threshold of 270 IU/l. The 34 patients with neoplastic obstruction (including 13 without jaundice) had more than twice the normal level of AP, and presented with at least one dilated bile duct in the biliary tree. Coronal scans of the main bile duct are fundamental in the diagnosis of the level of obstruction. It seems thus possible to affirm that US diagnosis of the biliary obstruction, together with high AP values (more than twice the normal), provides with reliable information as to the neoplastic nature of the biliary obstruction, even if jaundice is not present. PMID- 3041483 TI - The history of lupus erythematosus. From Hippocrates to Osler. AB - Hippocrates (460-375 BC) was the first to describe cutaneous ulcers under the heading of herpes esthiomenos. From what we can tell, Herbernus of Tours was the first to apply the term lupus to a skin disease in 916 AD. Following this, a number of terms including lupus, noli me tangere, and herpes esthiomenos were used to describe cutaneous ulcers. Willan (1757-1812) expanded the classification of skin diseases using the term herpes for vesicular diseases and lupus for destructive and ulcerative diseases of the face. The first clear description of lupus erythematosus was by Biett and was reported by his student Cazenave under the term erythema centrifugum in 1833. In 1846 Hebra, under the name of Seborrhea Congestiva described disc-shaped patches and introduced the butterfly simile for the malar rash. In 1851 Cazenave renamed erythema centrifugum, calling it lupus erythematosus and gave a classic description of discoid lupus erythematosus. In 1872 Kaposi subdivided lupus into the discoid and systemic forms and introduced the concept of systemic disease with a potentially fatal outcome. Hutchinson alluded to the photosensitive nature of the rash and may have provided the earliest description of what is now called annular subacute cutaneous lupus. In 1894 Payne used quinine in the treatment of patients with LE and postulated the presence of a vascular disturbance. In 1902, Sequira and Balean published a large series of patients with discoid and systemic LE and provided clinical and pathologic details of a young woman who died of glomerulonephritis. In 1904, Jadassohn published an exhaustive review of discoid and systemic LE, including clinical features and pathologic findings. Between 1895 and 1904 Sir William Osler published 29 cases of what was termed the erythema group of diseases. Perhaps his major contribution was to show that skin diseases could be accompanied by a variety of systemic manifestations. In retrospect most of his patients suffered from diseases other than SLE and it was only in his 1904 paper that two cases with SLE were described. He did not acknowledge this diagnosis in his cases and we share the viewpoint that his contribution to the study of SLE has been overemphasized. PMID- 3041485 TI - Cardiopulmonary manifestations of systemic lupus erythematous. AB - The cardiac and pulmonary manifestations of SLE are numerous. While the pleura and pericardium are most commonly affected, the myocardium and lung parenchyma can be the targets of life-threatening complications. Secondary processes (in particular, infection) must always be ruled out before attributing a cardiopulmonary manifestation to lupus. PMID- 3041484 TI - Renal disease in systemic lupus erythematosus. AB - Lupus nephritis is the archetype of diseases caused by the deposition of immune complexes. The characteristics which render only a portion of circulating immune complexes nephritogenic are not well delineated, but cat-ionic charge of antigen and/or antibody may contribute. Lupus nephritis is characterized by extreme diversity of clinical manifestations and pathologic features. Scrupulous monitoring of urinary sediment and renal function tests is necessary to identify renal involvement in a phase which is amenable to therapeutic intervention. Evaluation of renal biopsies also assists in development of indications for therapy. The pathology of lupus nephritis is classified primarily as mesangial, focal proliferative, diffuse proliferative, and membranous nephropathy. Indexes of activity and of chronicity provide a succinet description of the balance of reversible and irreversible disease. The activity index incorporates glomerular hypercellularity, karyorrhexis/fibrinoid necrosis, leucocyte exudation, hyaline thrombi, cellular crescents, and interstitial inflammation. The chronicity index incorporates glomerular sclerosis, fibrous crescents, tubular atrophy, and interstitial fibrosis. Delineation of the activity and chronicity indexes also facilitates the assessment of prognosis and the ordering of indications for therapy. PMID- 3041486 TI - Neuropsychiatric lupus. AB - Neuropsychiatric abnormalities in patients with systemic lupus have been recognized for more than a century. Although the prognosis of lupus has improved, involvement of the nervous system continues to be a major feature, with some abnormality recognized in 50 to 66 per cent of lupus patients. Diagnosis and therapy of neurologic disease remain the most difficult clinical challenges in the management of SLE. PMID- 3041487 TI - Epidemiology of systemic lupus erythematosus. AB - A major international study, properly designed and executed, could establish whether the population differences that seem to exist in prevalences of SLE are real and whether these differences, eg, in black and Chinese groups residing in different parts of the world, depend upon varied genetic constitution or whether they reflect environmental influences. The noninfectious environmental influences that should be considered are toxic and dietary ones. PMID- 3041488 TI - Pregnancy in systemic lupus erythematosus. AB - Issues concerning contraception, fertility, and pregnancy usually arise during a typical lupus patient's disease course. Pregnancy superimposed on established lupus may alter the course of the disease, and, conversely, lupus may affect the natural history of pregnancy. Two recently described autoantibody markers, anti SSA (Ro) and anticardiolipin, have provided new insights concerning fetal risks in these patients. Furthermore, they should lead to improved understanding of mechanisms of tissue injury and to new ideas about therapeutic interventions and/or prevention of pregnancy complications. PMID- 3041489 TI - Treatment. Corticosteroids and anti-inflammatory drugs. AB - Nonsteroidal anti-inflammatory drugs and corticosteroids are important elements in the therapeutic armamentarium for patients with systemic lupus. The choice of NSAID needs to be individualized, but with optimal usage NSAIDs can often be used to manage symptoms previously treated with corticosteroids. For serious disease manifestations, corticosteroids are the cornerstone of therapy. Maximization of clinical response and avoidance of side effects continue to be important management goals. Different dosage regimens, such as intravenous methylprednisolone pulse therapy, and adjunctive agents, such as cyclophosphamide, are of continued interest in severe and potentially life threatening disease. In addition, new nonsteroidal anti-inflammatory strategies including omega-3 series eicosanoids and new steroidal strategies including deflazacort and anti-glucocorticoids hold promise for continued improvement in the treatment on systemic lupus. PMID- 3041490 TI - Treatment. Disease-modifying therapies. AB - Many patients with systemic lupus can be treated effectively with antimalarials and nonsteroidal anti-inflammatory drugs without ever having to take systemic corticosteroids at all or for any significant length of time. On the other hand, some patients with life-threatening disease, active major organ disease, or intolerable corticosteroid toxicities should be given immunosuppressive drugs, plasmapheresis, and/or other therapies in addition to corticosteroids early in their disease course, before permanent, end-organ damage occurs and before the predictable serious and debilitating toxicities of prolonged, daily high-dose corticosteroids develop. Just as lupus patients now routinely undergo detailed serologic testing, it is conceivable that, in the future, routine determination of human leukocyte antigen (HLA) haplotypes45 and T-cell subsets88a will help define, at disease onset, those patients who are destined to have severe disease. Perhaps this knowledge, combined with a better understanding of the exact mechanisms of action of these disease-modifying therapies, will allow a more rational approach to the treatment of SLE. PMID- 3041492 TI - Outcome and prognosis in systemic lupus erythematosus. AB - We have attempted to assess the factors associated with prognosis in SLE, and to document the temporal changes in outcome, related not only to improvements in survival but to the emergence of an increased prevalence or morbidity, related to disease manifestations, complications of treatment, and co-morbid conditions. PMID- 3041491 TI - Pathogenesis of systemic lupus erythematosus. AB - SLE is a syndrome characterized by diverse clinical signs and symptoms, including rash, serositis, nephritis, central nervous system disease, thrombocytopenia, and leukopenia. These and other manifestations are present in patients with immune abnormalities indicative of B cell hyperactivity: hypergammaglobulinemia, increased amounts of antibodies reactive with self-determinants (characteristically including nuclear antigens), and increased numbers of circulating antibody-producing cells. It is this immune hyperactivity, inclusive of anti-self responses, which groups the patients with diverse clinical manifestations into a single syndrome called lupus. PMID- 3041493 TI - Musculoskeletal manifestations of systemic lupus erythematosus. AB - The musculoskeletal system is involved in nearly all patients with SLE. Transient arthralgias or arthritis are common and, in some patients, there is a progression to rheumatoid-like nonerosive hand deformities (Jaccoud's syndrome). The major disabling type of joint disease in lupus is articular osteonecrosis, often induced by high-dose corticosteroids. Rare forms of musculoskeletal manifestations of lupus include spontaneous tendon rupture, crystalline arthropathies, subcutaneous calcifications, and inflammatory myopathy. PMID- 3041495 TI - [Acquired immunodeficiency syndrome (AIDS) and public health]. PMID- 3041494 TI - [Efficiency of cefmetazole and cefoxitin in the treatment of sepsis caused by gram-negative bacteria]. PMID- 3041497 TI - [Neonatal pulmonary pathology]. AB - The authors study the detailed symptomatology of the respiratory distress syndrome in the new born, and review the principal pulmonary pathologies encountered in the neonatal period. For each one the physiology, epidemiology and the clinical and paraclinical aspects were studied as well as those factors concerning the evolution and prognosis. Therapy was also discussed and for certain disorders the future prospects in this field. PMID- 3041496 TI - [Physiology and physiopathology in the development and maturation of the antenatal lung]. AB - The term "pulmonary maturation" is reserved for the process of surfactant production. The term "pulmonary development" covers the overall phenomena of the growth of pulmonary structures and their differentiation. Pulmonary development is divided schematically into five stages: the embryonic stage up to the fifth to the seventh week of gestation, then the four stages of the foetal period. These four stages are: 1) The pseudo-glandular stage up to the seventeenth week during which the bronchi and the extra-acinar vessels are formed; 2) The canalicular stage characterised by the birth of the acinus, the differentiation of the cells and the beginning of surfactant synthesis to the end of this stage; 3) The saccular and alveolar stages. The saccular stage begins between the 24th and 26th week of gestation and at this stage the foetus is viable but exposed to the risk of neonatal respiratory distress on account of lack of surfactant. The alveoli are formed before the end of pregnancy. The factors controlling development and pulmonary maturation are still poorly understood. Cellular, particularly epithelial and mesenchymal interactions appear to play a major role in all the stages of development. Hormonal interactions play a determining role in pulmonary maturation. The effects on development and/or pulmonary maturation of pharmacological agents, hormones administered to the mother (e.g. opiates, glucocorticoids, aminophylline, beta-sympathomimetics, indomethacin and tobacco...) and maternal pathology are described as far as possible within the limits of our current knowledge. This information comes above all from experimental studies, thus the conclusions obtained are difficult to apply to newborn humans. PMID- 3041498 TI - [Pregnancy and the respiratory function]. AB - The respiratory function of pregnant women is changed for more than one reason. There is a mechanical effect due to the increase in the uterine volume and the elevation of the diaphragm. However, there are only modest functional consequences, because the pulmonary volumes are only little changed, with the exception of a reduction in the functional residual capacity. Bronchial permeability is unaltered due to the balancing of constrictors (mechanical, hypocapnia) and dilators (hormonal influences). As regards pulmonary haemodynamics, the hypovolaemia of the pregnant woman has no repercussions on pulmonary vascular pressure. Gas exchange and alveolar-capillary diffusion are normal or even improved as a consequence of chronic hyperventilation. The latter is the most important functional change, sometimes expressed as a sensation of dyspnoea. The origin of this hyperventilation relates to the diminution of the threshold of sensibility in the respiratory centres to CO2 due to the effect of raised progesterone levels. PMID- 3041499 TI - [Respiratory problems related to general anesthesia and artificial respiration in the pregnant woman and during childbirth]. AB - General anaesthesia in pregnancy is still responsible for a significant morbidity and mortality. The most common and most serious complications are respiratory secondary to changes induced by pregnancy. These are dominated by hypoxia during difficult intubation and inhalation of gastric contents. Their incidence could be largely reduced by the extensive use of regional local anaesthesia. PMID- 3041500 TI - [Chronic respiratory diseases and their decompensation during pregnancy (asthma excluded)]. AB - There are relatively few observations on the influence of pregnancy in chronic respiratory illness, excluding asthma and interstitial pneumonia. Chronic airflow obstruction only presents at a relatively advanced age, past the menopause. Chronic respiratory failure due to restrictive lung disorders appears well tolerated as long as the vital capacity is greater than one litre. Mucoviscidosis has been the object of more detailed studies since the management of this disorder now raises hopes of survival compatible with pregnancy. PMID- 3041501 TI - [Interstitial pathology and pregnancy]. AB - Pregnancy is an important factor in the outcome of interstitial pneumonias. The outcome for sarcoidosis and rheumatoid disease is favourable as a rule, and these disorders have little influence on the progress of the disease. Idiopathic pulmonary fibrosis or fibrosis complicating scleroderma or dermatopolymyositis requires careful management considering the frequent complications at foetal level. During systemic disorders, recourse to corticosteroid-cyclophosphamide regime is possible from the second trimester. Drug induced pneumonias remain rare, do not influence pregnancy and most often only require the arrest of the incriminated drug. PMID- 3041503 TI - [Antibiotics during pregnancy and breast feeding: consequences for the treatment of respiratory infections]. AB - Respiratory infections are the second most frequent cause for antibiotic prescriptions during pregnancy, after genito-urinary infections. Overall, antibiotics are relatively innocuous. The following should be avoided: the tetracyclines, cotrimoxazole, chloramphenicol, metronidazole and the quinolones. Aminoglycosides should be administered controlling the plasma level. The beta lactones (principally the penicillins) and the macrolides sold in France, are without any danger. Nevertheless, the rise in distribution volume and the overall physiological changes which accompany the developing pregnancy, particularly in the third trimester lead to a diminution in the serum concentration of these antibiotics and imply an adaptation, often a doubling, of the therapeutic dose administered. From the epidemiological data concerning the organisms involved in the respiratory infections, nearly the totality of extra-hospital infections may be cured by macrolides or penicillins (ampicillin). During more serious infections (nosocomial, or the immuno-depressed) the maternal prognosis should take precedence, adjusting the antibiotic to the organism, before the toxic risk to the child. All the antibiotics are excreted in the mother milk, but in very small quantities; they are generally destroyed in the digestive tract of the child so that the risk of any secondary effect during lactation is minimal. PMID- 3041502 TI - [Tuberculosis and pregnancy]. AB - Tuberculosis is neither more frequent, nor more serious in pregnant than non pregnant women. The risks for the child are threefold: a doubling in the mortality level on account of the illness in the mother if she is not treated; a risk of toxicity linked to the anti-tuberculous drugs and a risk of tuberculous infection at birth. Isoniazid (INH) and ethambutol have a weak toxicity. These two antibiotics can be prescribed during pregnancy (after confirming the absence of Vitamin B6 deficiency in the mother). Rifampicin is teratogenic in high doses in animals, but epidemiological studies do not reveal any notable risk in man. For prudence it is only prescribed in the first trimester of pregnancy, in confirmed cases of tuberculosis. The data on the teratogenicity of pyrazinamide is insufficient and it should not be used in pregnancy. Thus the treatment of tuberculosis in pregnancy will be rifampicin + isoniazid + ethambutol (the ethambutol being stopped after two months and the isoniazid and rifampicin after 9 months of treatment). At the moment of confinement, if maternal tuberculosis is confirmed bacteriologically at the time of microscopy, chemoprophylaxis will be started in the new born with isoniazid, in a dose of 5 mg/kg until the mother is bacteriologically negative on microscopic examination, the new born should then be vaccinated with BCG. If the treatment of the mother is correctly prescribed and followed breast-feeding is possible and no isolation of either mother or child is necessary. The amount of antibiotic that passes in the mothers milk is minimal and such specific nourishment should not be dispensed with if the treatment is necessary. PMID- 3041504 TI - [Growth and aging of lung: special reference to pulmonary diseases]. PMID- 3041505 TI - [Persistent atrial standstill]. PMID- 3041506 TI - [Cardiac preservation: current status and unsolved problems]. PMID- 3041507 TI - [Bronchial asthma and psychogenic factor]. PMID- 3041508 TI - Hyperplasia and carcinoma of the endometrium. AB - Published criteria for classifying endometrial hyperplasias and their differential diagnosis from well-differentiated endometrioid-type adenocarcinoma are reviewed, and the author's own interpretations and recommendations are presented. Also discussed are the subjects of misdiagnosis of hyperplasia and carcinoma, premalignant significance of hyperplasia, and prognostic significance of hyperplasia in patients with carcinoma patients. PMID- 3041510 TI - Pure mesenchymal neoplasms of the uterine corpus: selected problems. AB - The first part of this review includes a discussion of some issues in decision analysis as it applies to uterine mesenchymal tumors, as well as a discussion of some of the morphologic features and histochemical techniques that may be used to determine whether a mesenchymal tumor is demonstrating smooth muscle or endometrial stromal differentiation. Also included in the first part are caveats that the pathologist should observe before interpreting mesenchymal tumors of the uterus. The second part of the paper is devoted to morphologic diagnostic criteria and differential diagnosis. PMID- 3041509 TI - Squamous differentiation in carcinoma of the endometrium: a critical appraisal of adenoacanthoma and adenosquamous carcinoma. AB - Although squamous differentiation within endometrial carcinomas has long been recognized by pathologists, its biologic significance has been the subject of continued debate. While some authors have found a worsened prognosis for women who have tumors with squamous elements, others have reported the prognosis to be better than conventional endometrial adenocarcinomas. Persisting confusion and disagreement about the use of the terms adenoacanthoma and adenosquamous carcinoma have complicated the issue. In this article we review the literature on the pathogenesis of squamous differentiation in the endometrium and discuss the histologic features, prevalence, and biologic behavior of adenocarcinoma with squamous differentiation. We conclude that keratin is a constituent of normal and neoplastic endometrial epithelial cells, and that overt squamous differentiation occurs by a mechanism that is currently unknown. Squamous differentiation is present in about 25% of endometrial adenocarcinomas, a frequency that appears to have been constant for the past 50 years. The squamous component of endometrial carcinomas may histologically appear benign, malignant, or indeterminant, and in the majority of instances closely parallels the differentiation of the glandular component. Endometrial adenocarcinomas with malignant-appearing squamous elements usually have poorly differentiated glandular components and have a prognosis identical to that of poorly differentiated adenocarcinoma without squamous differentiation. Endometrial adenocarcinomas with benign-appearing squamous elements are usually associated with well-differentiated glandular components and have a prognosis identical to that of typical well-differentiated adenocarcinoma. Some endometrial adenocarcinomas contain foci of squamous differentiation that appear neither clearly benign nor malignant. These often are associated with moderately differentiated glandular components; the prognosis for these women is not yet clearly defined. At the present time we are unable to attribute any prognostic significance to the presence of squamous differentiation in endometrial carcinomas. Because of the confusion and semantic arguments that revolve about the use of the terms adenoacanthoma and adenosquamous carcinomas, we recommend that those terms be abandoned and be replaced by the single term adenocarcinoma with squamous differentiation. As for any other endometrial tumor, the pathologist should provide information on histological grade, depth of myometrial invasion, and presence of vascular involvement or spread to the cervix in order to guide the gynecologist in determining appropriate therapy. PMID- 3041512 TI - Invasive procedures in the diagnosis of pneumonia. AB - The etiology of respiratory infections can be elusive despite the recent advances in diagnostic technology. Thereby, the clinician needs a systematic approach for a definitive early diagnosis. This review presents the pros and cons of various invasive procedures in order to select the most appropriate diagnostic method. Expectorated sputum, the important initial step in community-acquired pneumonia, is unreliable in complex pneumonias, mainly because of colonization of the oropharynx. Even though blood cultures are frequently negative, they provide a precise diagnosis and should be obtained in undiagnosed pneumonias. Transtracheal needle aspiration has few false-negative results. However, its use has decreased because of the high frequency of false-positive cultures and risk of serious complications. Bronchoscopy provides direct access to both the bronchi and parenchyma for visualization and sampling. Plugged telescoping catheter brush, used safely in different clinical settings, has good sensitivity in identifying the pathogen, but the specificity varies with the underlying status of the patient. Because of upper airway contamination, bronchial washings are only slightly better than expectorated sputum. A newer technique, transbronchial needle aspiration, is, thus far, no improvement over the plugged telescoping catheter brush. In the immunosuppressed patient, bronchoalveolar lavage has excellent diagnostic accuracy for opportunistic infections. The accuracy increases with the addition of transbronchial biopsy. Transthoracic needle aspiration gives decisive information with low false-positive results. With the ultrathin needle, the complications decrease. Renewed interest in thoracoscopy guided biopsy has demonstrated a high diagnostic accuracy with low complication rate. One procedure, a combination, or improvements of these procedures may reduce the need for open lung biopsy. Nevertheless, an open lung biopsy furnishes the best specimen for making a histological and microbiological diagnosis, although controversy exists regarding any improvement in survival rates. PMID- 3041511 TI - Using the microbiology laboratory in the diagnosis of pneumonia. AB - Pneumonia/influenza is one of the top ten leading causes of mortality in the United States each year. The identification of the etiologic agent responsible for lower respiratory tract infection plays an important role in the proper management of this clinical problem. The specimens submitted for evaluation are obtained in diverse ways and include expectorated sputum, material from transtracheal and bronchoscopic procedures, pleural fluid and lung aspirates, and biopsy of actual lung tissue. Processing of material can include stained smears, aerobic and anaerobic cultures, and special processing techniques for fungal, viral, Pneumocystis carinii, Legionella, mycobacterial, and mycoplasma identification. Modifications of smear preparation techniques and application of the new DNA probe technology are providing the opportunity for rapid microbiologic testing of clinical specimens with increased sensitivity and specificity, often obviating the need for invasive diagnostic procedures. Laboratory methodology is continually undergoing technological change, and optimal care of the patient with pneumonia requires close cooperation between the attending physician and the clinical laboratory. PMID- 3041513 TI - The serodiagnosis of nonpneumococcal bacterial pneumonia. AB - The diagnosis of pneumonia is often difficult when only staining and culture of clinical specimens are used. Testing for antigens or antibody in serum is an alternative method. Many different tests have been described or are currently being used for detection of bacterial pathogens that cause pneumonia, and many more are being developed. It is often difficult to decide which tests to use or how to interpret their results. Frequently, the sensitivity and specificity of each test is not well characterized. A practical approach to serodiagnosis of bacterial pneumonias other than pneumococcal pneumonia is presented. In addition to the sensitivity, specificity, and interpretation of standard serologic tests, the proper role of newer serologic techniques is discussed for each bacterial agent. PMID- 3041514 TI - Diagnosis of pneumococcal pneumonia. AB - Pneumococcal pneumonia presents peculiar problems to the diagnostician. It is at once the most common form of community-acquired bacterial pneumonia and simultaneously the most difficult to document microbiologically. Bacteremia, empyema, meningitis, or septic arthritis due to S pneumoniae unmistakably verifies this bacterium as the cause of a coexistent pneumonia; this coexistence fortunately occurs infrequently. The diagnostic dilemma arises in the less sick patient. While recognizing the common presence of pneumococci in the oropharynx of healthy individuals, we give undue credence to S pneumoniae cultured from sputum obtained by expectoration. At the same time, pneumococci are frequently not found in cultures of sputum obtained from patients with confirmed bacteremic disease. More invasive techniques (transtracheal aspiration, protected bronchoscopic catheter, lung needle aspiration) are too complex, dangerous, or both for routine use. Attempts to detect pneumococcal antigen in blood, sputum, or urine by modern immunologic techniques give promise of avoiding the problems of either contamination or lack of bacteriologic growth. However, they have not yet been evaluated in sufficiently large groups with pneumonia of independently determined bacterial etiology to calculate test sensitivity and specificity. At the present time then, the careful clinician will use all the epidemiologic and clinical evidence at hand, including a careful Gram's stain and culturing of sputum, blood, and other sources, to arrive at the most likely etiology. The probabilities must be weighed in light of the imprecision of current laboratory confirmation and modified by clinical course. Choice of antimicrobial therapy still favor penicillin for patients with community pneumonia severe enough to warrant hospitalization, despite ominous trends in multiple resistance of S pneumoniae. PMID- 3041515 TI - Diagnosis of nosocomial pneumonia. AB - Nosocomial pneumonia occurs in 0.6% of hospitalized patients. The usual causative agents are gram-negative bacilli, Staphylococcus aureus, Streptococcus pneumoniae, and anaerobic bacteria. In immunocompromised hosts, the differential diagnosis also includes fungi, mycobacteria, viruses, Nocardia, and Pneumocystis carinii. Important risk factors for the development of nosocomial pneumonia include prolonged mechanical ventilation, thoracic or upper abdominal surgery, altered mental status, underlying immunosuppression, chronic obstructive pulmonary disease, and the use of antacids or histamine type 2 blockers. Colonization of the oropharynx and tracheal secretions with gram-negative aerobic bacteria is common in hospitalized patients with or without pneumonia. The diagnosis of nosocomial pneumonia is usually based on the clinical features of dyspnea, cough, fever, purulent sputum production, new pulmonary infiltrates, hypoxemia, and leukocytosis. However, the clinician must recognize that the presence of these features is neither sensitive nor specific in the diagnosis of nosocomial pneumonia. Microbiologic diagnosis is also difficult because blood cultures are usually negative, and cultures of tracheal secretions, although usually sensitive, are not specific. Invasive procedures may prove useful, but most have yet to be studied in large groups of patients with nosocomial pneumonia. PMID- 3041516 TI - Diagnosis of pulmonary fungal infections. AB - The diagnosis of pulmonary fungal infections is reviewed. An overview of the major endemic and opportunistic fungal infections is followed by a brief discussion of the available diagnostic tools. These include: (1) direct microscopic examination of secretions or body fluids; (2) histopathologic examination of tissue biopsies; (3) cultures of secretions, body fluids, or ground-up tissue; and (4) immunological tests ("fungal serologies"). In the final section, a current diagnostic approach is given that identifies the diagnostic tests more likely to be helpful for each infection. PMID- 3041517 TI - Parasitic pneumonia. AB - With few exceptions, the parasitic pneumonias most commonly encountered in the Western Hemisphere are diseases of compromised hosts; patients with AIDS are at particular risk. Pneumocystis carinii pneumonia occurs eventually in 80% of AIDS patients; bronchoalveolar lavage is quite sensitive in establishing this diagnosis. Toxoplasma gondii pneumonia, seen most often in the patient with AIDS, is characterized by multisystem involvement. Strongyloides stercoralis infection is endemic in the Southeastern United States. Pulmonary strongyloidiasis is seen in patients receiving glucocorticoids or chemotherapy, and in patients with AIDS or other causes of T cell dysfunction. Larvae may be seen on Gram's stains or wet mounts of sputum. Ascaris and hookworm infections may present with pulmonary infiltrates and eosinophilia during the larval migration phase. Dirofilaria, Paragonimus, and Entamoeba histolytica involvement of the lung are less common and require a good epidemiologic history and clinical suspicion for diagnosis. PMID- 3041518 TI - The role of plain films, CT, tomography, ultrasound, and percutaneous needle aspiration in the diagnosis of inflammatory lung disease. AB - Traditionally, plain film chest radiography has been the mainstay in the roentgenographic evaluation of infectious disease in the chest. Plain film tomography has augmented the chest radiograph in the detection of cavitation and in evaluation of the bronchial tree. Newer imaging modalities including ultrasound, which evaluates the pleural space, and computerized tomography (CT), which examines the lung parenchyma, mediastinum, pleura, and chest wall, have further aided in diagnosis. Finally, percutaneous aspiration of the lung or pleural space under radiologic guidance now allows specific diagnoses to be made more frequently and with fewer complications. PMID- 3041519 TI - [Insulin therapy optimized]. PMID- 3041521 TI - [Current findings in thyroid autoimmunity]. PMID- 3041520 TI - [Computerized tomography-guided thoracic puncture in the diagnosis of pulmonary nodular lesions. Apropos of 5 year's experience]. PMID- 3041522 TI - [Pregnancy and thyroid diseases]. PMID- 3041523 TI - [Thyroid-hormone resistance syndromes]. PMID- 3041524 TI - [Marfan disease and cardiovascular pathology]. PMID- 3041525 TI - [Cutaneous metastases of visceral neoplasms. Review of the literature apropos of 2 anatomo-clinical reports]. PMID- 3041526 TI - [Cardiological monitoring of patients treated with doxorubicin]. PMID- 3041527 TI - [Urinary lithiasis in children. Etiology, examination and urological treatment]. PMID- 3041530 TI - [From Harvey to Furchgott and de Bold]. PMID- 3041531 TI - [The vascular endothelium as mediator and target organ of cardiovascular diseases]. PMID- 3041532 TI - [The renin-angiotensin system in hypertension and heart insufficiency: current aspects in the use of ACE inhibitors]. PMID- 3041528 TI - Iron status in athletes. An update. AB - As more studies are done on the iron status of athletes, the significance of apparent iron deficiency remains controversial. Do observed changes in iron status in athletes indicate an actual iron deficiency or a physiological response to exercise? Iron replacement would clearly be indicated if an iron deficiency was present but would not be necessary or effective if the observed changes were simply a physiological response. There is agreement that serum ferritin and haemoglobin decrease with some exercise conditions and that some indicators of haemolysis, such as serum haptoglobin and bilirubin, change in response to exercise. Expansion of plasma volume and the shift of iron storage from bone marrow to the liver could support the claim that the apparent reduced iron status parameters occurring with exercise are misleading. Countering this concept are studies in athletes which demonstrate dietary iron intake deficiencies and blood loss in the gastrointestinal and urinary tract. Iron deficiency is common in the general population, particularly in women. Therefore, continued monitoring of iron status in athletes appears justified in the face of present knowledge. Replacement therapy, when iron deficiency is apparent, is recommended. PMID- 3041533 TI - [Silent myocardial ischemia]. PMID- 3041529 TI - Blood pressure behaviour during physical activity. AB - Aerobic exercise is currently being recommended in addition to pharmacological therapy for lowering blood pressure levels in hypertensive patients, i.e. in subjects whose resting blood pressure levels exceed 145/90 mm Hg. On the other hand competitive sports are generally contraindicated in hypertensives, who are thought to be at increased risk of morbidity or mortality from their blood pressure levels. The present knowledge of blood pressure behaviour during isotonic physical activity is almost wholly based on the results obtained by means of the ergometric tests. Several maximal and submaximal exercise protocols have been introduced, but none has proved to be superior for diagnostic purposes. There is general agreement that the systolic blood pressure increase determined by isotonic exercise usually ranges from 50 to 70 mm Hg in both normotensive or hypertensive subjects. Diastolic blood pressure shows only minor changes in the normotensives, while in the hypertensives it tends to substantially increase because of their inability to adequately reduce their peripheral resistance. This mechanism may also explain the delay shown by the hypertensives in reaching pre exercise blood pressure values during the recovery. On average diastolic blood pressure increases to a greater extent during bicycle ergometry than during treadmill, while no differences in exertional systolic blood pressure have been observed between the 2 tests. The results of several studies indicate that the blood pressure response to isotonic exercise is a marker for detection of hypertension earlier in the course of the disease, while resting blood pressure is still normal. According to some authors it is also of value in predicting future hypertension in individuals with borderline pressure levels. There are no conclusive data on the effect of training on blood pressure response to exercise. The majority of the published studies report small exertional pressure reductions after conditioning, which would merely reflect the reduction in resting blood pressure. Vasodilation greatly influences the exercise-induced rise in blood pressure; in fact the exertional pressor increase is blunted when the test is preceded by an adequate warm-up session. Isometric effort is thought to be contraindicated in hypertensive subjects, as it causes a pronounced increase not only of systolic but also of diastolic pressure. Mean blood pressure is, however, increased to the same extent by isotonic and isometric exercise, even though minor discrepancies have been reported by some authors.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3041535 TI - [Congenital diaphragmatic hernia]. PMID- 3041534 TI - [Long-term therapy of type II and type IV hyperlipoproteinemia using beclobrate]. PMID- 3041537 TI - [On the track of oxygen]. PMID- 3041536 TI - [Serratia marcescens pneumonia: a rare postoperative complication in cancer patients. Apropos of a case]. PMID- 3041538 TI - Nocturnal oscillations in plasma renin activity and REM-NREM sleep cycles in humans: a common regulatory mechanism? AB - To establish the strength of the relationship between the nocturnal oscillations in plasma renin activity (PRA) and the sleep stage patterns, 42 PRA profiles from blood collected at 10-min intervals and the concomitant polygraphic sleep recordings were analyzed. In all cases, PRA curves exactly reflected the pattern of sleep stage distribution. When sleep cycles were complete, PRA levels oscillated at a regular 100-min period, with a strong spectral density. Declining PRA levels always coincided with REM sleep phases and increasing levels with NREM sleep phases. More precisely, peak levels corresponded to the transition from deep sleep stages toward lighter ones. The start of the rises in PRA generally marked the transition from REM sleep to stage 2. For incomplete sleep cycles, PRA curves reflected all disturbances and irregularities in the sleep structure. Spontaneous and provoked awakenings blunted the rise in PRA normally associated with NREM sleep, which indicates that disturbing sleep modifies the renin release from the kidneys. These results suggest that a common mechanism within the central nervous system controls both PRA oscillations and the REM-NREM sleep alternation. PMID- 3041539 TI - [Vulvar intra-epithelial neoplasms. Development of ideas and facts]. AB - The authors report their personal experience concerning the recent increase in the frequency of VINs and demonstrate how the mean age of the patients has decreased. They advocate "vulvoscopy" and guided biopsy for screening. VIN I and II may be treated with local chemotherapy and/or laser vaporisation. VIN III must be treated surgically (superficial tailored resection). PMID- 3041540 TI - [Management of a pregnant woman with Streptococcus group B]. AB - The relative rarity (1 to 5 cases for 1,000 births) of neonatal infections secondary to B Streptococcus, the epidemiological characteristics of this germ, especially the unstable vaginal carriage, make it difficult to select a therapeutic approach. Systematic screening of B Streptococcus and the treatment of all carriers or only of high-risk patients, present several practical problems, are complex to implement but the cost/benefit ratio seems however acceptable. Prophylactic intrapartum antibiotic treatment of known carriers of B Streptococcus does not seem debatable any longer, at least the treatment of those presenting other risk factors: premature delivery, premature rupture of the membranes, fever occurring during delivery. Today, the best prophylaxis of neonatal infections seems to be the intrapartum antibiotic treatment (ampicillin) resulting in a spectacular decrease of the frequency of neonatal contamination. PMID- 3041541 TI - [Active management of labor in Dublin: what is it 15 years later?]. AB - Every obstetrician, in his readings, must have come across a report of article by K. O'Driscoll, and could not remain indifferent (surprise, incredulity, even anger...) in front of astonishing results: 5% of caesarean sections, 5 to 6% of forceps deliveries, combined with a recruiting worthy of any large university center and creditable neonatal results which are accurately analysed. In view of the difficulty and understanding of the basis of "active management of labor", advocated by this team and described as the main reason of its success, the author tried to understand, while staying in this department, the highlights of its daily obstetrical practice. In this detailed report of the activities of this irish obstetrical team, the author wanted to put the results back in their context and broadly outline the organization of this department and the management of labor, in order to evaluate the significance of the figures reported in numerous international publications and too often not easy to analyse for uninformed readers. PMID- 3041543 TI - [Importance of serologic detection of syphilis at the time of blood donation (prevention of transfusion syphilis)]. PMID- 3041542 TI - [Probable association of HLA-DR5 with bullous pemphigoid]. AB - HLA typing was performed in 35 French Caucasoids with bullous pemphigoid and compared with 160 healthy controls. 47 HLA antigens were characterized by a lymphocytotoxicity micromethod. Analysis of the results only reveals one statistically significant difference: an increased incidence of HLA-DR5, which reaches 51.43% in patients versus 22.42% in controls, with P = 0.0007 and Pc = 0.0329. Several bullous dermatosis are associated with various HLA-DR antigens. These data suggest a direct role of HLA-DR molecules in the constitution of these autoimmune disease. An abnormal expression of DR products on some skin cells membrane would permit the presentation of a non self peptide, accumulated in skin cells, to helper T lymphocytes. An heteroimmunization against the non self peptide could lead to lesion of self cells. This peptide perhaps derives from food protein. PMID- 3041544 TI - [Prevention of transfusion malaria]. PMID- 3041546 TI - Reactivity of tumor cells in malignant effusions with a panel of monoclonal and polyclonal antibodies. AB - A panel of 13 mouse monoclonal antibodies (mAb) and 1 rabbit polyclonal antibody was tested for reactivity with tumor cells in 26 effusions obtained from patients with carcinoma of ovary, breast, or mesothelioma, using an immunoperoxidase staining reaction. Specific staining of tumor cells but not reactive mesothelial cells was demonstrated with some of the mAb in the panel. In 4 of 26 effusions no evidence of malignancy was obtained after routine cytological staining, but this was reversed on the basis of immunoperoxidase staining of tumor cells with the mAb in the panel. Serial effusions were evaluated in 4 patients during the course of chemotherapy, allowing an assessment of effect of therapy on the antigenic characteristics of the tumor cells. In another 4 patients, the results of immunoperoxidase staining of effusions were compared with those obtained by applying the same antibody panel to solid tumor nodules. There was a tendency to develop changes in the pattern of reactivity during therapy, and the pattern of reactivity was more restricted in tumor nodules than in effusions. One of the mAb in our panel (2G3) was consistently shown to produce strong staining of a high proportion of tumor cells in effusions and tumor nodules from patients with ovarian or breast cancer and may be of value in immunocytochemical diagnosis and therapy of epithelial malignancies. PMID- 3041547 TI - The carcinoembryonic antigen gene family: structure, expression and evolution. AB - The molecular cloning of carcinoembryonic antigen (CEA) and several cross reacting antigens reveals a basic domain structure for the whole family, which shows structural similarities to the immunoglobulin superfamily. The CEA family consists of approximately 10 genes which are localized in two clusters on chromosome 19. So far, mRNA species for five of these genes have been identified which show tissue variability in their transcriptional activity. Expression of some of these genes in heterologous systems has been achieved, allowing the localization of some epitopes. The characterization of a CEA gene family in the rat and a comparison with its human counterpart has been utilized in the development of an evolutionary model. PMID- 3041545 TI - A clinical and biochemical evaluation of etretinate in rheumatoid arthritis. AB - Etretinate (Tigason; Roche), which is effective in the treatment of psoriatic arthritis has immunomodulating activity in vivo. We have therefore assessed this drug in an open clinical and biochemical assessment of 24 weeks duration in rheumatoid arthritis. The treatment dose was 1.0 mg/kg/day for the first 4 weeks reducing to 0.5 mg/kg/day thereafter. There was a modest clinical improvement though this only reached statistical significance for joint circumference at 12 and 16 weeks (P less than 0.05). Biochemical improvement only reached levels of statistical significance for IgM at week 16 (P less than 0.01). Eight out of 15 patients had discontinued the drug because of side-effects by week 12 and only three out of 15 patients showed individual improvement by week 24. Some biochemical parameters (ESR) worsened. These results suggest only modest clinical efficacy and use of the drug in rheumatoid arthritis is likely to be curtailed by unacceptable side-effects. The improvement in biochemical variables that occurs when the drug is used in psoriatic arthritis does not occur in rheumatoid arthritis. PMID- 3041548 TI - [Bibliography of the scientific publications of the staff of the Bialystok Medical School 1981-1982]. PMID- 3041549 TI - Special issue devoted to the 25th anniversary of the Revue Roumaine de Medecine, Serie de Medecine Interne. PMID- 3041550 TI - Hyperlipoproteinemia, hemostatic variables and thromboatherosclerosis. AB - A concept is gradually being established that certain forms of hyperlipoproteinemia are apt to be accompanied by disturbances of the hemostatic balance favouring fibrin deposition and the incorporation of microthrombi into the arterial wall. Author's clinical and laboratory observations emphasized that endogenous hypertriglyceridemia is often accompanied by raised plasma levels of fibrin stabilizing factor XIII, fibronectin and fibrinolytic inhibitors rendering the fibrin clots more resistant to fibrinolysis and more readily attached to the subendothelial layers of the vessel wall. Increased activity of vitamin K dependent clotting factors partially counterbalanced by a rather high level of antithrombin III were also detected. Since endogenous hypertriglyceridemia is characterized by an accelerated synthesis and turnover of lipoproteins it may be suggested that this process somehow stimulates the hepatic production of the above-mentioned hemostatic variables. The subsequent alteration of the hemostatic balance is compatible with the localized character and the slow progression of thromboatherosclerosis. PMID- 3041551 TI - Coronary atherosclerosis: where are we now? AB - An attempt is made to present and summarize recent results in atherosclerosis research which may aid to a better understanding of atherogenesis, progression of atherosclerotic lesions and occurrence of myocardial ischemia. Based on selected data from the available literature and the authors' experience, the review focuses the attention on: the atherogenic role of hemodynamic stresses; the onset and fate of early coronary atherosclerotic lesions; the development of lesions of possible clinical significance; the concept of "critical stenosis"; the view that atherosclerosis is a hyperplastic and/or neoplastic disease. PMID- 3041552 TI - Advances in neurological therapy (a review). PMID- 3041553 TI - Immunopathology and humoral autoimmunity in chronic active hepatitis. A critical review. PMID- 3041554 TI - A criterion for completeness of vagotomy based on basal and vagally stimulated gastric acid secretion after esophagectomy or proximal gastric vagotomy. AB - The variation in basal acid secretion was determined in 10 patients after resection of the esophagus, an operation resulting in a total transection of all vagal nerves to the abdomen. After recording basal acid secretion over a 3-h period, a 15-min modified sham feeding procedure was performed, and the acid output was studied for an additional hour. The mean basal acid output +2 SD was 0.27 mmol/15 min. The difference between the highest and lowest recorded 15-min output--that is, the oscillation of basal acid output--was calculated for each patient. The mean oscillation of basal acid output +2 SD was 0.58 mmol/15 min. Vagal stimulation accomplished by sham feeding produced no significant increase in acid output above this level. The variation in basal acid secretion was also investigated in 20 duodenal ulcer patients after proximal gastric vagotomy. These patients were insulin-negative and remained asymptomatic during a 7- to 10-year follow-up study. Shortly after the vagotomy, measurement of basal acid secretion over 3 h showed a mean basal acid output +2 SD of 0.58 mmol/15 min. The mean oscillation of basal acid output +2 SD was 0.66 mmol/15 min. On the basis of the oscillation in basal acid secretion after complete vagotomy we propose a new criterion for completeness of vagotomy, namely a response to physiologic vagal stimulation which does not exceed the lowest basal level by more than 0.6 mmol/15 min. PMID- 3041555 TI - Clinical efficacy of endoscopic injections of OK-432 in the treatment of gastric cancer. AB - A total of 48 patients with gastric cancer were randomly assigned to receive either endoscopic injections of OK-432 plus systemic treatment with intravenously administered 5-fluorouracil and intradermally injected OK-432 (group A) or systemic therapy alone (group B). Morphologic improvement occurred in 7 of the 22 patients in group A. In one patient the tumor mass disappeared completely. In group B morphologic improvement occurred in only 2 of the 26 patients. The survival rate throughout the 24-month period was significantly (P less than 0.01) higher in group A than in group B patients. Thus, local administration of OK-432 by endoscopic injection is effective and is recommended for the treatment of patients with advanced gastric cancer. PMID- 3041556 TI - Glucagon and insulin for the treatment of hepatic failure in dimethylnitrosamine intoxicated rats. AB - When rats received dimethylnitrosamine every 24 h until death, plus hormone treatment after the first 24 h, the survival was enhanced between 100 and 140 h compared with the control rats, with attenuated derangements of prothrombin time and serum albumin levels at 120 h. In rats given a single dose of dimethylnitrosamine, hepatic DNA synthesis peaked at 48 h. The synthesis was increased after hormone treatment when started immediately, but not when delayed for 24 h. Hormone treatment for 3 days starting 24 h after a single dose of dimethylnitrosamine produced a rapid normalization of decreased hepatic protein content on day 9, although hepatic DNA content was not affected. These results suggest that this treatment is effective for hepatic failure, and the promotion of restoration of liver function is a contributing factor to its effect, in addition to the stimulation of hepatocyte proliferation. PMID- 3041557 TI - Pancreatic polypeptide response to insulin in duodenal ulcer. Different levels in accordance with ulcer activity and its response to treatment. AB - Pancreatic polypeptide is said to be a marker of vagal tone in duodenal ulcer. To determine whether pancreatic polypeptide levels are related to the course of duodenal ulcer, we studied acid and pancreatic polypeptide responses to insulin in 80 patients with duodenal ulcer disease: 40 with unoperated duodenal ulcer and 40 with proximal vagotomy. Data were analysed in accordance with the presence of an ulcer (active disease) and, when present, in accordance with the ulcer healing on medical treatment (cimetidine, 1 g/day for 4 weeks). In both groups acid and pancreatic polypeptide responses to hypoglycaemia were slightly correlated (r = 0.38) (p less than 0.05). The basal pancreatic polypeptide level was higher in patients with active disease than in those with inactive disease, who had a basal level similar to that of normal subjects of the same age range. Like the insulin stimulated acid secretion, the pancreatic polypeptide response to insulin hypoglycaemia was higher in patients with active disease than in those with inactive disease (p less than 0.05): 26.1 +/- 3.9 versus 20.1 +/- 4 nmol/l/120 min, respectively, in unoperated patients and 34.8 +/- 2.2 versus 24.3 +/- 2.5 nmol/l/120 min, respectively, after proximal vagotomy. In active disease the pancreatic polypeptide response to insulin hypoglycaemia was higher in subjects whose ulcer did not heal further after cimetidine therapy than in those whose ulcer did. These data suggest that the pancreatic polypeptide response to insulin is an indicator of duodenal ulcer activity and is related to the treatment efficacy. These relationships are partly mediated by increased vagal tone. PMID- 3041559 TI - Cell-mediated immunity to Plasmodium falciparum infection: evidence against the involvement of cytotoxic lymphocytes. AB - Blood mononuclear cells (PBMC) recognizing soluble malaria antigens (SPag) are present in the peripheral blood of individuals clinically immune to malaria, and they proliferate after exposure to such antigens. To test whether these cells have effector activity against Plasmodium falciparum, we stimulated PBMC from malaria-immune donors by SPag and purified protein derivative (PPD) in culture for 7 days. The PBMC were then co-incubated with P. falciparum for 48 h, and parasitaemia was determined by microscopy. Parasite growth was only significantly impaired after incubation with PBMC stimulated by either SPag or PPD in the presence of immune serum. Studies on subpopulations of PBMC indicated that the inhibitory cells resided among the adherent cell fraction. Furthermore we tested PBMC for cytotoxic activity against P. falciparum-infected autologous or heterologous erythrocytes. Experiments were done both in the absence and the presence of immune serum. Neither fresh PBMC nor PBMC activated by SPag or PPD for 7 days prior to assay were cytotoxic, indicating that cytotoxic T cells, natural killer (NK) cells, and K cells did not possess cytotoxic activity directed against parasitized erythrocytes. These data support the hypothesis that activated monocytes are the most important effector cells in the peripheral blood of malaria immune individuals. PMID- 3041558 TI - Omeprazole or ranitidine in the treatment of reflux esophagitis. Results of a double-blind, randomized, Scandinavian multicenter study. AB - One hundred and fifty-two patients with endoscopically verified erosive and/or ulcerative esophagitis entered a double-blind, randomized study comparing 20 mg omeprazole given once daily and ranitidine 150 mg twice daily. The efficacy and safety of 4 to 8 weeks' treatment were studied. Macroscopic healing of esophagitis was defined as complete epithelialization of all esophageal erosive and/or ulcerative lesions. One hundred and forty-four patients completed the first 4 weeks of treatment in accordance with the protocol. The healing rate was 67% in the omeprazole group and 31% in the ranitidine group (p less than 0.0001). The corresponding figures after 8 weeks' treatment were 85% and 50%, respectively (p less than 0.0001). The higher healing rate for omeprazole was also accompanied by a significantly faster and more substantial improvement in reflux symptoms. In the patient's own overall evaluation of symptoms, these had resolved in 51% of the omeprazole-treated patients already at the end of the 1st week of treatment, compared with 27% of those given ranitidine (p = 0.009). Both omeprazole and ranitidine were well tolerated, and there were no adverse events or clinically significant changes in the laboratory values attributable to the trial medication. PMID- 3041560 TI - Characteristics of soluble tumour-derived proteins that inhibit natural killer activity. AB - We have previously shown that various benign and malignant natural killer (NK) resistant monolayer cells inhibit endogenous human NK activity, probably by reducing the secretion of cytotoxic factors from the effector cells. The nature of the molecules responsible for the inhibition has been unclear. In this study we show that phosphate-buffered saline (PBS) extracts of ovarian cystadenocarcinoma tissue and normal uterine smooth muscle strongly inhibit NK activity. Fractionation of tumour extracts by gel chromatography revealed major inhibitory activity in the Mr range 160,000-180,000, and other weaker inhibiting activities in the Mr ranges 50,000-70,000 and 20,000. The active material of Mr range 160,000-180,000 was adsorbed on anion exchange chromatography column at neutral pH and physiologic NaCl concentration, and it was eluted by 0.31-0.34 M NaCl. The inhibitory molecule was sensitive to proteolysis. No relation of this compound to immunoglobulins or trypsin and urokinase inhibitors was detected. The unfractionated extract inhibited NK activity apparently by the same mechanism as the monolayer target cells, i.e. by reducing the secretory capacity of effector cells. The data strongly suggest that the NK-inhibiting compounds described in this work are involved in the inactivation of NK cells by intact monolayer cells. PMID- 3041561 TI - Beta-haemolytic group A, B, C and G streptococcal septicaemia: a clinical study. AB - 87 beta-haemolytic streptococcal septicaemias in adult patients during 1979-86 in a university hospital were reviewed. 25% were caused by group A streptococcus, 17% by group B, 14% by group C and 44% by group G streptococcus. 67% of the septicaemias due to group B streptococcus were nosocomial, whereas the group A, C or G septicaemias were in most cases community-acquired. Alcoholism was the most common underlying disease in group A (32%) and malignancy in group G streptococcal septicaemias (45%). The most common origin and focus of infection in group A, C and G streptococcal septicaemias was the skin. The total mortality in beta-haemolytic streptococcal septicaemias was 20%, higher in septicaemias caused by group A (32%) and group B (33%) than by group C (17%) and group G (8%) streptococci. Nevertheless, there were more patients in group G streptococcal septicaemias with severe underlying diseases than in other groups of beta haemolytic streptococci. The present data seem to indicate that a septicaemia due to group G is a more benign disease than a septicaemia due to group A streptococcus. PMID- 3041562 TI - Erythema nodosum and conjunctivitis triggered by enteritis due to Salmonella typhimurium. AB - We describe a patient, a 59-year-old woman, who developed erythema nodosum and conjunctivitis after enteritis due to Salmonella typhimurium. The patient was HLA B27 negative and had circulating immune complexes, as determined by the conglutinin-binding enzyme immunoassay. Immunopathogenesis of the extraintestinal findings is discussed. PMID- 3041563 TI - Acyclovir crystalluria. AB - Acyclovir crystals in the urine of a 28-year-old woman are described. The patient received intravenous acyclovir for herpes encephalitis. The crystalluria was accompanied by impairment of the renal tubules as evidenced by increased urinary beta-2-microglobulin excretion. Consecutive series of 60 patients on oral acyclovir and 20 patients on intravenous acyclovir were studied in order to define the frequency of acyclovir crystalluria. None of the patients studied had crystalluria, and none elevated serum creatinine levels. It is suggested that acyclovir crystalluria might be an early sign of acyclovir crystal nephropathy. PMID- 3041564 TI - Intrathecal application of drugs for muscle hypertonia. AB - The literature regarding the intrathecal use of morphine, baclofen, and midazolam to treat spasticity is reviewed. Nine patients with significant spasticity due to different etiologies were treated. Morphine and midazolam decreased spasticity but did not change the patient's functional status. Baclofen improved patient status, but was associated with significant CNS depression in two cases. PMID- 3041565 TI - The concept of inner cerebral trauma. AB - The pattern of "inner cerebral trauma" is a morphological and functional pathological correlate of biomechanical conditions at the event of severe closed craniocerebral injury of acceleration-type, if the traumatizing forces act in the direction of the longest diameter of the skull. The lesions are characteristically localized in the "centro-axial" regions of the brain involving most frequently: corpus callosum, septum pellucidum, fornix, tela chorioidea, peri- and para-ventricular zone, infundibulobasal region and cingulum; this pattern also includes lesions of the hippocampal area, upper brainstem, pontocerebellar complex, and parasagittal areas of the cerebrum. This pattern includes lesions, which are from the onset "primary irreversible" as well as "primary reversible" lesions, which spread apart from the "epicentres" of the primary irreversible damage and are in principle more diffuse but still within the basic pattern of the main predilection. This is the basis of the process of "traumatic cerebral disease". The final size and scope of the pathological process of the "traumatic cerebral disease" depend on a number of secondary factors, which may allow the normal process of healing to keep the tissue changes within the initial scope of "primary irreversible damage", or may enhance a progressive process of turning the "reversible" traumatic damage into the irreversible lesions. PMID- 3041566 TI - Behavioural abnormalities in head injured patients. AB - This paper reviews the psychological deficits found after head injury. The deficits are very broadly of two types involving cognitive and behaviour disturbances. The main features of the two types of disturbances are described; and the natural history, prediction, functional impact, and rehabilitation of the disturbances are reviewed. Key features of good clinical evaluation of patients are identified. PMID- 3041567 TI - Head injuries and restorative neurology. PMID- 3041569 TI - The dento-gingival junction as seen with light microscopy and scanning electron microscopy. AB - The purpose of this paper is to review the anatomical relationship of the Dento Gingival Junction as seen in the human dentition. The junction is described under light microscopy and then reviewed as seen in the SEM with the author's unpublished findings. The authors' material was derived from extracted human teeth with remaining marginal gingival tissue. The specimens were fixed with 2% glutaraldehyde in 0.15M sodium cacodylate buffer (pH 7.2) for 24 h. The specimens were then washed and freeze-fractured in Freon 113 using liquid nitrogen. Afterwards they were processed by freeze-drying or CPD methods, coated with gold, and placed in the scanning electron microscope (SEM) for viewing. These specimens demonstrated the presence of numerous Sharpey's fibers at the cemental surface. A large number of fibrils intermingled with the fibers to produce a dense mass of tissue. Junctional epithelium, with the adjacent homogeneous dental cuticle was demonstrated. Plaque deposits on the tooth surface extended to a cell-free zone. Morphological detail viewed with SEM and light microscopy are compared. PMID- 3041568 TI - Bioapplication of colloidal gold in microbiological immunocytochemistry. AB - Microbiological organisms are an ubiquitous group of animals encompassing bacteria, viruses, protozoa, algae and fungi. They are adapted for survival in many diverse habitats and exert a profound effect on man and his environment. Colloidal gold electron immunocytochemistry is a useful technique for studying these organisms and may be applied in several ways. The post-embedding technique is used to detect internal antigens, whilst the pre-embedding technique is employed for the detection of external antigens. In contrast the immuno-negative stain technique is applied to detect antigens on structures such as viruses or bacterial pili which may be dried down onto an electron microscope grid and immunolabelled in situ. In addition the immunoreplica technique allows the examination of cell surfaces for the appearance of antigens. Together these techniques have yielded valuable information concerning microbiological organisms. PMID- 3041570 TI - The role of scanning electron microscopy in periodontal research. AB - During recent years a great amount of research has led to a better understanding of the etiology, pathogenesis and pattern of progression of periodontal diseases. Scanning electron microscopy (SEM) has contributed to this improvement, mainly with respect to histology of periodontal tissues, the description of the morphology and distribution of bacteria on the exposed root surface, analysis of the host-parasite interactions on the gingival pocket wall, and morphological evaluation of root treatment. This review deals with all these topics. Unusual types of SEM research are also described and discussed. Uncommon sample preparation techniques for SEM in periodontal research are described. SEM in periodontal research should be of great application in the near future. Cathodoluminescence, back-scattered emission and immunolabelling techniques will be formidable tools in this field of dentistry. PMID- 3041571 TI - A review of methodology and quantification in dental microwear analysis. AB - Dental microwear analysis is a method of inferring oral events (primarily food processing and aspects of masticatory biomechanics) from microscopic abrasion patterns retained on the enamel surfaces of teeth. Although some qualitative pattern differences may be easily distinguishable, most of the significant results produced thus far have derived from quantified studies of SEM images of occlusal enamel. It often goes unnoticed by readers of microwear reports who are not themselves specialists that microwear analysis is essentially a statistical method, not a visual one. In this review of current techniques and methods, several problems in current approaches are detailed. It is noted that feature definition can have significant effects on ultimate pattern differentiation. Sampling bias is also a major concern, as most microwear studies are carried out on samples which are very small. Compounding this are the effects of magnification level choices, and the effects of SEM instrumentation on feature visibility. Finally, the interpretation of pattern differences requires careful attention to comparisons of within-group and between-group variability. PMID- 3041572 TI - A review of dental microwear and diet in modern mammals. AB - Recent work has shown that microscopic wear patterns on teeth may yield insights into variations in diet and tooth use in modern and prehistoric mammals. This paper presents a review of dental microwear and diet in modern mammals, plus a discussion of topics for further research. To date, incisor and molar microwear have been examined, although there are far fewer studies of the former. Facilitated by the use of high-resolution casts and scanning electron microscopy, analyses have ranged from: qualitative to quantitative, low magnification to high magnification, and experimental studies to comparative studies of museum collections. Results are encouraging and may lead to further insights into a variety of topics including food processing and dental microstructure. PMID- 3041573 TI - Overview of physical studies of bulk water in biopolymers. AB - The Symposium of which this paper is a part, "The State Of Water In The Cell", examines evidence which indicates that the bulk water of the cell has physical properties that are different from those of free water in a dilute aqueous solution. The heterogeneity of environments of water within cells makes direct application of physical studies difficult. However, the major physiologic effect of an ordered state of bulk cell water is solute exclusion, and this is also demonstrated by simple polymer solutions. Such systems have provided an opportunity to show that such water has motional correlation times within an order of magnitude of those of free water, and to model other unique properties of bulk cell water. PMID- 3041575 TI - Clinical applications of scanning electron microscopy and energy dispersive X-ray analysis in dermatology--an up-date. AB - Dermatological papers comprising scanning electron microscopy (SEM) and energy dispersive X-ray (EDX) analysis data published 1983 through 1986 in international journals are reviewed, as an update to our 1984 paper on Clinical applications of scanning electron microscopy and X-ray microanalysis in dermatology. The present paper not only deals with a review of recent publications in this area but also presents the application of microincineration to hair and cryosectioned freeze dried skin specimens. Examples of the increased contrast obtained in hair cross sections are presented and a discussion on the feasibility of microincineration at analysis of hair and skin cross sections is given. Particle probe analysis (EDX: energy dispersive X-ray analysis and PMP: proton microprobe analysis) as applied to hair and skin samples are presented with stress put on the proton probe analysis. The complementarity of EDX and PMP is demonstrated and future applications are suggested. PMID- 3041574 TI - Solute exclusion by polymer and protein-dominated water: correlation with results of nuclear magnetic resonance (NMR) and calorimetric studies and their significance for the understanding of the physical state of water in living cells. AB - According to the polarized multilayer (PM) theory of cell water proteins with their backbones fully extended and their NHCO groups directly exposed to bulk water, polarize water in multilayers. Experimental testing of the theory led to a new understanding of the uniqueness of gelatin, due to its permanently maintained fully extended conformation and its ability to polarize the bulk phase water in multilayers with reduced solubilities for solutes in a size dependent manner ("size rule"). Other models which behave like gelatin are urea-denatured proteins, synthetic polymers like polyethylene oxide (PEO), and polyvinylpyrrolidine (PVP), but not native proteins. NMR studies showed that the majority of water molecules dominated by these polymers does indeed suffer rotational (and translational) motional restriction as predicted by the PM theory. In conjunction with ultra-high frequency dielectric studies but particularly quasielastic neutron scattering of both model systems (e.g., PEO) and living cells (i.e., brine shrimp cysts and frog muscle), this finding offers confirmation of the PM theory of living cell water and model systems. Studies of the freezing point depression showed that the presence of as much as 50% of native proteins had no effect on the freezing point of water while inclusion of gelatin, PEO, etc., caused concentration-dependent lowering of the freezing temperature. These findings demonstrate the key role of polarized water in the phenomena of freezing point depression and the unusual ice forms seen in living cells. PMID- 3041576 TI - Arterial elastin as seen with scanning electron microscopy: a review. AB - All large arteries contain elastin, collagen, and muscle which can be seen with light microscopy and transmission electron microscopy. Elastin forms an internal elastic lamina (IEL) in all arteries, but also forms multiple fenestrated sheets in the media of the aorta and other large arteries. The fenestrations in the media are larger than those in the IEL. The adventitial elastin is more fibrous and often contains tubular elastin surrounding vasa vasorum when prepared by removing all non-elastin by placing the aorta in 0.1 N NaOH at 70-75 degrees C for five hours. The fenestrations are larger near branches and in an experimentally created poststenotic dilatation. Atherosclerosis appears associated with both new elastin formation in early atherosclerosis and elastolysis in late disease. PMID- 3041577 TI - [Nuclear medicine: current aspects and future]. AB - A survey of the range and progress of nuclear medicine procedures is presented and the accuracy of the modalities is described. The priority in nuclear medicine diagnosis has shifted from proof of the non-functioning nodule to localization of biochemical lesions with the aid of new bioindicators and single photon and positron emission computer tomography. Diagnosis has extended to subcellular regions with the introduction of immunoscintigraphy as well as enzyme and receptor scintigraphy. This is a new dimension to which Swiss nuclear medicine specialists have contributed a great deal. PMID- 3041578 TI - [Optimal possibilities in pediatric radiology]. AB - The feasibility of compliance with the WHO guidelines on safe and economical use of pediatric radiology is discussed. The easiest way of reducing risks and costs is to dispense with unnecessary investigations; this is possible without harm in many diagnostic fields. Replacement of more invasive techniques, especially by ultrasonography, has made an outstanding contribution to pediatric radiology. PMID- 3041579 TI - [Echography of superficial structures]. AB - Sonography is nowadays a routine procedure when gallstones or a tumor of an abdominal organ are suspected. More recently with advance in technology it has become possible to image superficial structures. Modern high resolution realtime machines show soft tissue structure and movements. Duplex machines show simultaneously tissue structure and blood flow by gated Doppler. The most recent machines show coloured blood-flow. The superficial structures studied by sonography are: the abdomen in pediatrics, the brain and medulla under the age of 6 months, the eye, the breast, the testes, the small parts of the neck and the extremities, non-ossified skeleton. Imaging of these structures is quite well known. Diagnostic value of simultaneous blood-flow studies has to be explored. Miniaturization of ultrasound transducers allows applications during surgery and endoscopy. Practical use of these procedures has to be defined. Because of all that work in progress ultrasound is now in touch with most fields of somatic medicine. Basic knowledge of ultrasound has to be integrated in the training of the future doctors to the same extent as radiology PMID- 3041580 TI - [Whole body magnetic resonance imaging]. AB - Magnetic resonance imaging (MRI) has a growing number of applications in the whole body area. Its role cannot be properly considered without comparing it to the diagnostic capabilities and availability of sonography and the high standard of newest-generation CT. MRI is an accepted diagnostic modality for diseases involving the pelvis and the musculoskeletal system. It is equal or superior to other modalities in examination of the neck, the cardiovascular system, the upper abdomen and the retroperitoneum. PMID- 3041581 TI - [Medical radiology--today and tomorrow]. AB - Medical radiology comprises diagnostic radiology, nuclear medicine and radio oncology. Digital radiography, sonography, computed tomography, magnetic resonance and percutaneous techniques mark the present development of radiodiagnosis. Current advances in nuclear medicine are emission computed tomographic techniques and biochemical radiopharmaceuticals. Computerized planning, combination therapy and particular types of ionizing radiation are the main ongoing developments in radiation oncology. Teaching and research have a decisive bearing on the future of medical radiology. PMID- 3041582 TI - [Clinical applications of magnetic resonance spectroscopy]. AB - In this overview the physics of magnetic resonance spectroscopy (MRS) are briefly discussed. The biochemical pathways which can be investigated by MRS in physiological and pathological states are presented. The potential clinical applications of this method to the musculoskeletal system, heart, brain, liver and kidney are examined. PMID- 3041583 TI - [75 years of the Swiss Roentgen Society. A look backward and forward]. PMID- 3041584 TI - [The Doppler palm test]. AB - In 384 hands the patency of the arterial arch was investigated by means of both the Allen test and the Doppler palmar arch test as developed by the authors. While the Allen test is purely subjective and gives a high proportion of false results especially in shocked patients, the Doppler palmar arch test is based on two objective criteria: 1. Demonstration of reversed flow in the distal section of the radial artery on proximal compression. 2. Demonstration of pulsatile flow in the radial thumb artery even on compression of the radial artery. The Doppler palmar arch test eliminates all negative aspects of the Allen test and is therefore definitely more reliable than the latter. It is easy to perform for investigators having little experience with Doppler sonography. The time consumed is not greater than with the Allen test. PMID- 3041585 TI - [The value of ultrasound Doppler pulse curve parameters in the assessment of stenotic arterial diseases]. AB - By means of a biophysical model of the pulse wave propagation in arterial vessels the impact of haemodynamic changes on the Doppler pulse curve was studied. The variation of the haemodynamic parameters of the model gives information about the diagnostic meaning of the pulse curve parameters in respect of stenotic alterations. PMID- 3041586 TI - [Sonographic assessment of vascular bypasses in vitro]. AB - 13 bypasses and 8 patches were examined by ultrasound in vitro with a 3.5 MHz and a 10 MHz probe. Whereas only 6 bypasses were assessable by sonography, all patches could be demonstrated in vitro. In these cases, sonography could be accomplished with either of the two frequencies used. The 10 MHz probe was superior for imaging structural details. Immediate postoperative sonography of such grafts, which cannot be demonstrated accurately in vitro, is not reliable. PMID- 3041587 TI - [Ultrasound imaging of the bile ducts and assessment of gallbladder kinetics in healthy newborn infants]. AB - The gallbladder and the biliary tract of 85 neonates were sonographically examined, in the fasting state. The gallbladder was detectable in all neonates and showed a measurable contraction. Two types of biliary contraction could be observed: a concentric contraction of length, depth and width (concentric type) or a contraction of depth and width (collapse type). The grade of postprandial contraction was independent of the level of serum bilirubin. Furthermore, sonographic criteria were established allowing the differentiation of the bile ducts and their discrimination from hepatic arteries. PMID- 3041588 TI - [Computerized sonographic imaging of tongue motility using pseudo-3 dimensional reconstruction]. AB - A new method for analysis and documentation of tongue movements was used in 24 healthy volunteers. This method enables direct digitalization of the information obtained from sequences of B-Mode pictures. Besides of allowing to further digital evaluation of the findings, it offers the facility of "pseudo 3D" reconstruction of the tongue movements occurring on swallowing. The method appears to be of particular value for therapy control in neurological and phoniatric patients. PMID- 3041590 TI - Mrs Isabella Elder, LL.D., pioneer and philanthropist. AB - Mrs Isabella Elder contributed much during her lifetime to further the education of women of all ages and of all levels of attainment. She gifted North Park House to Queen Margaret College in Glasgow as a home in 1884. With her encouragement a medical school for women in Glasgow was opened in Queen Margaret College in 1890. The standard was high and when women were admitted to the Universities in 1892, the first Queen Margaret medical students were able to graduate in 1894. She pioneered a system of teaching home economics in Govan, assisted the development of a district nursing service and built the Elder Cottage Hospital. PMID- 3041589 TI - [Documentation of scanning array position in sonography of the extremities]. AB - A simple system is presented for the documentation of probe position in real time sonography of extremities. Manufacturers of equipment are requested to offer frequently required text lines in a menu option. A programme of this kind is presented for use in orthopaedics. PMID- 3041591 TI - Intervacuole exchange in the yeast zygote: a new pathway in organelle communication. AB - A new pathway of vesicle traffic between organelles has been identified. The vacuoles (lysosomes) of Saccharomyces cerevisiae zygotes rapidly exchange their contents at a specific point in the cell cycle. With the use of fluorescence microscopy, "tracks" were observed that connect the original parental vacuoles to the newly forming bud vacuoles. These observations suggest that vacuole-derived vesicles rapidly move along the tracks in both directions, equilibrating vacuole contents. This rapid vesicle movement may be responsible for vacuole formation in newly developing cells. PMID- 3041592 TI - Escherichia coli aspartate transcarbamylase: the relation between structure and function. AB - The x-ray structures of the allosteric enzyme aspartate transcarbamylase from Escherichia coli have been solved and refined for both allosteric forms. The T form was determined in the presence of the heterotropic inhibitor cytidine triphosphate, CTP, while the R form was determined in the presence of the bisubstrate analog N-phosphonacetyl-L-aspartate. These two x-ray structures provide the starting point for an understanding of how allosteric enzymes are able to control the rates of metabolic pathways. Insights into the mechanisms of both catalysis and homotropic cooperativity have been obtained by using site directed mutagenesis to probe residues thought to be critical to the function of the enzyme based on these x-ray structures. PMID- 3041593 TI - Breast cancer-associated pS2 protein: synthesis and secretion by normal stomach mucosa. AB - The human pS2 gene is specifically expressed under estrogen transcriptional control in a subclass of estrogen receptor-containing human breast cancer cells. The pS2 gene encodes an 84-amino acid protein that is secreted after signal peptide cleavage. The distribution of pS2 protein in normal human tissues was studied with antibodies to pS2; pS2 was specifically expressed and secreted by mucosa cells of the normal stomach antrum and body of both female and male individuals. Moreover, no estrogen receptor could be detected in these cells, indicating that pS2 gene expression is estrogen-independent in the stomach. The function of the pS2 protein in the gastrointestinal tract is unknown. However, the pS2 protein is similar in sequence to a porcine pancreatic protein that has been shown to inhibit gastrointestinal motility and gastric secretion. PMID- 3041595 TI - [Effects of systemic antibiotic therapy on the pathogen spectrum and development of resistance in chronic osteomyelitis]. PMID- 3041596 TI - [Early results of meniscal sutures in the so-called degeneration zone. Clinical aspects, arthrography and meniscus computerized tomography]. PMID- 3041594 TI - Wound macrophages express TGF-alpha and other growth factors in vivo: analysis by mRNA phenotyping. AB - The presence of macrophages is required for the regeneration of many cell types during wound healing. Macrophages have been reported to express a wide range of mitogenic factors and cytokines, but none of these factors has been shown in vivo to sustain all the wound-healing processes. It has been suggested that transforming growth factor-alpha (TGF-alpha) may mediate angiogenesis, epidermal regrowth, and formation of granulation tissue in vivo. Macrophages isolated from a wound site, and not exposed to cell culture conditions, expressed messenger RNA transcripts for TGF-alpha, TGF-beta, platelet-derived growth factor A-chain, and insulin-like growth factor-1. The expression of these transcripts was determined by a novel method for RNA analysis in which low numbers of mouse macrophages were isolated from wound cylinders, their RNA was purified and reverse-transcribed, and the complementary DNA was amplified in a polymerase chain reaction primed with growth factor sequence-specific primers. This single-cell RNA phenotyping procedure is rapid and has the potential for quantification, and mRNA transcripts from a single cell or a few cells can be unambiguously demonstrated, with the simultaneous analysis of several mRNA species. Macrophages from wounds expressed TGF-alpha antigen, and wound fluids contained TGF-alpha. Elicited macrophages in culture also expressed TGF-alpha transcripts and polypeptide in a time-dependent manner after stimulation with modified low-density lipoproteins and lipopolysaccharide endotoxin, which are characteristic of the activators found in injured tissues. PMID- 3041597 TI - [Fixation of a cruciate ligament replacement. Experimental evaluation of mechanical stability in cadaver knees]. PMID- 3041598 TI - The role of chemotherapy in diffuse aggressive lymphomas. AB - Diffuse aggressive lymphomas are curable by combination chemotherapy in advanced stages, with 30% to 60% of all treated patients alive and disease free 2 to 5 years from the end of treatment, depending on the program used. The fact that combination chemotherapy can cure this disease has led, since 1975, to several new directions in therapy such as (1) the development of a variety of new combination chemotherapy programs, using older drugs more aggressively, adding additional drugs to each program, or manipulating the scheduling of drugs; (2) the close scrutiny of study results to identify risk factors that influence the ability to attain a durable complete remission; and (3) the use of combination chemotherapy in early stage disease instead of, or with, radiotherapy (RT). The recently developed aggressive programs have been reported to produce results superior to the original studies, but because important prognostic factors have only recently been identified, there remains some uncertainty about the significance of the reported differences between studies. Also, dose intensity, a poorly controlled treatment variable in all studies, has recently been identified as a significant factor affecting outcome. Schedule variation, while reported to exert some independent effect on outcome, appears to influence results by its effect on dose intensity. Regardless of the uncertainty over the ideal program for patients with advanced diffuse aggressive lymphoma, the results in early stage disease have uniformly favored the use of combination chemotherapy as the primary treatment, with or without subsequent RT, depending on the quality of the response to drugs, reaffirming the invariable inverse relationship between cell number and curability by chemotherapy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3041599 TI - Results of recent salvage chemotherapy regimens for lymphoma and Hodgkin's disease. AB - From 1981 to 1983, 208 patients with recurrent or refractory lymphoma were treated with methylglyoxal-bis-guanylhydrazone (methyl-GAG), ifosfamide, methotrexate, etoposide (MIME). The complete remission (CR) rate was 24% and CR plus partial remission (PR) was 60%. Response was higher for aggressive than for indolent cell types. Median duration of CRs was 16.5 months and median survival of all patients was 9 months. In view of the in vitro synergism between cisplatin and high-dose cytarabine, we recently designed the DHAP regimen, which consists of cisplatin, 100 mg/m2 IV over 24 hours; cytarabine, 2 g/m2 IV over two hours every 12 hours for two doses; and dexamethasone, 40 mg orally daily for 4 days. There were 28 of 90 (31%) CRs and 22 of 90 (24%) PRs. Median duration of CR is 15 months; median survival of all patients is 6 months. The major toxicities have been infection (31%) and moderate to severe renal dysfunction (20%). In contrast to MIME, response rates did not differ significantly between aggressive and indolent cell types. A high-dose regimen (CBV) consisting of cyclophosphamide, 6 g/m2; carmustine, 300 mg/m2; and etoposide, 250 mg/m2 daily for 3 days followed by autologous bone marrow transplant (ABMT) has been successfully used to treat 62 patients with Hodgkin's disease recurrent or refractory after mechlorethamine, vincristine, procarbazine, prednisone, doxorubicin, bleomycin, vinblastine, dacarbazine (MOPP/ABVD)-like regimens. A CR rate (47%) has been observed; 83% of these CRs remain alive and free of disease with a median follow-up of 19 months. This regimen appears to have curative potential in 40% of all cases and in 60% of cases treated after the first or second relapse.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3041600 TI - Treatment strategies for Hodgkin's disease. AB - Over the past 2 decades, treatment of Hodgkin's disease has evolved considerably through innovations in the management of various stages. The impact of various treatments on the 5-, 10-, and 15-year results is being balanced against delayed morbidity, such as organ damage and second malignancies, produced by the intensity of therapy or the prolonged delivery of given drugs. The results of clinical trials performed during the past decade have allowed us to reconsider the various prognostic variables that can be used in the treatment strategy. The major unfavorable prognostic factors are represented by bulky disease, multiple extranodal sites, systemic B symptoms, age greater than 60 years, lymphocyte depleted histology, male sex, and progressive disease during chemotherapy. In patients with early disease after surgical staging, the aim of current therapy is to provide a high cure rate within a short period and with limited morbidity. In patients with advanced Hodgkin's disease, the treatment strategy is to achieve durable complete remission in most cases through effective, full-dose, multidrug regimens at the expense of acceptable morbidity. Subtotal or total nodal radiotherapy (RT) induces a 10-year cure rate ranging from 70% to 85% in stages I and II with no bulky lymphoma. In patients with bulky disease and all three systemic symptoms, comparable results can be achieved with primary chemotherapy followed by RT. Currently, stages IIIA and IIIB disease are often managed with combined treatment modalities, although comparable results can be obtained with intensive chemotherapy alone.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3041602 TI - The development of surfactant replacement therapy and the various types of replacement surfactants. AB - This overview has been intended to ease the reader who is not familiar with the surfactant replacement field into this area. The discussion of various surfactants should help in the clarification of terminology use by different authors and provides a perspective for the following articles, which deal more completely with many of the issues raised in this article. PMID- 3041601 TI - Autologous bone marrow transplantation in the treatment of malignant lymphoma and Hodgkin's disease. AB - High-dose chemotherapy, both with and without radiotherapy, was pioneered in the treatment of acute leukemia in relapse with allogeneic transplantation, exploiting the steep dose-response curve characteristic of some hematologic neoplasms. Extension to the malignant lymphomas was strengthened by early success in Burkitt's lymphoma and in syngeneic transplantation for lymphoma. The optimal regimen (BACT [carmustine, cytarabine, cyclophosphamide, 6-thioguanine]) and setting are still under investigation for the various grades of lymphomas. The early published experience demonstrated a low ultimate "cure" rate when transplantation was performed in advanced, bulky, and refractory disease. A survey of published reports up to 1986 showed only 16 of 112 long-term, disease free survivors when autologous bone marrow transplantation (ABMT) was performed in refractory relapse as opposed to 33 of 53 for patients transplanted in second or subsequent remission or in first partial remission. Refractoriness to conventional-dose chemotherapy (no response or progressive disease) cannot be salvaged in the majority of cases. Bone marrow involvement complicates the use of ABMT and may require in vitro elimination with monoclonal antibodies/complement or cytotoxic chemicals. The Dana-Farber Cancer Institute experience shows that the former in vitro treatment does not inhibit bone marrow grafting. When selection criteria for ABMT are applied in drug-sensitive relapse, 50% to 60% long-term, disease-free survival may be expected. Definition of poor prognostic factors in large cell lymphoma may identify patients for ABMT as consolidation of first remission. The issue of marrow purging is unsettled at the present time.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3041603 TI - Biophysical behavior of lung surfactant: implications for respiratory physiology and pathophysiology. AB - The major emphasis of this article has been the complex, multicomponent system of surfactants that are required for proper pulmonary mechanics and function in the mammalian lung. Although LS was discovered over 30 years ago, and soon after was linked directly with neonatal RDS, it has taken a significant time for researchers to develop a fundamental understanding of the pulmonary surfactant system, and its actions and roles in respiratory physiology. Nonetheless, knowledge about LS has increased greatly over the past decade, and it is now clear that exogenous surfactant replacement therapy for infants with RDS provides a substantial clinical advantage for these patients. Indeed, the therapy is life saving in many very small premature infants, and as experience accrues, and therapy is optimized, this advance is clearly a major step forward in neonatology. Perhaps the most prominent theme that has been presented throughout the discussion here is that pulmonary surfactant research must take advantage of interdisciplinary descriptions and cross-correlations for accurate and rapid progress. One positive feature of prior work on lung surfactant replacement and RDS is that its difficulty has forced investigators toward a level of understanding that is sound enough to extend LS research into related fields, such as lung injury and ARDS. These areas have their own complications, including a much more diverse pathology and injury progressions than found with neonatal RDS. In fact, if defining the role of lung surfactant in ARDS (and developing replacement therapy for it) had been the goal of investigators before considering neonatal RDS, it is difficult to imagine a positive outcome. The situation now, however, is one where it is realistic to think of recognizing when and how LS effects will occur in different ARDS lung injuries, so that surfactant replacement will have the best opportunity to help mitigate their progressive pathology. In dealing with ARDS, it is well to remember that there are a variety of complicating factors, since lung injuries vary with animal age and species, and according to the level and duration of exposure. For example, in the hyperoxic injury described in the previous section there was clearly significant LS involvement. However, had animals been subjected to a lower level of oxygen (eg, 60%), even for a comparatively long time (eg, 21 days), the entire pathologic pattern would have been altered, as demonstrated, for example, by Holm et al.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3041604 TI - Structure and function of pulmonary surfactant proteins. AB - In summary, the isolation and characterization of three surfactant proteins have considerably changed our understanding of the nature of pulmonary surfactant and its metabolism. The isolation of the cDNAs and genes encoding the proteins and the elucidation of their structure now makes possible the generation of surfactant proteins for further study and for therapy of surfactant deficient states. Artificial surfactant consisting of appropriately modified human proteins can be produced by recombinant DNA technology. Mixed with appropriate synthetic phospholipids, these proteins may be useful for the treatment of infants with hyaline membrane disease. Significant progress has been made in identifying and characterizing these three more abundant proteins; however, many questions regarding the functions of numerous other proteins present in surfactant (whether produced by Type II cells, Type I cell, Clara cell, or others) remain unanswered. Knowledge of the factors controlling developmental expression of surfactant phospholipids and protein as well as the molecular basis of the interactions among the surfactant proteins and phospholipids may lead to new strategies for therapy of hyaline membrane disease. Major questions regarding the site and nature of posttranslational modification of the surfactant proteins, their sites of assembly with lipids, and their precise roles in the metabolism of surfactant in vitro remain to be clarified. It is hoped that answers to these questions will facilitate the identification and design of appropriate therapy of infants and adults with pulmonary disease associated with surfactant deficiency. PMID- 3041605 TI - Immunologic consequences of exogenous surfactant administration. AB - In conclusion, we have shown that human surfactant is immunogenic and that circulating surfactant-antisurfactant immune complexes are detectable in the plasma from infants and in adults with RDS. We found these immune complexes regardless of whether exogenous surfactant was used in the individual treatment regimen. These immune complexes do not yet seem to cause disease in the short term. Long-term effects, if any, are unknown. Indications for surfactant replacement therapy in neonatal RDS are clear. Trials of exogenous surfactant are just beginning in adult RDS, and potential immunogenicity will be of even greater concern in these patients. In all such situations, potential for side effects must be balanced against therapeutic efficacy and the gravity of the disease. Our data indicate that surfactants, particularly heterologous surfactants, are potent immunogens. One cannot assume that using homologous or heterologous surfactants in patients with RDS will always be immunologically innocuous. Nonetheless, based on present data, moderately long-term follow-up (2 to 4 years), we are encouraged by our observation that no selective adverse effects attributable to human surfactant have been recognized, yet mortality from RDS in infants less than 30 weeks has been nearly cut in half. PMID- 3041606 TI - Metabolism of endogenous surfactant and exogenous surfactants for replacement therapy. PMID- 3041607 TI - Surfactant for the treatment of respiratory distress syndrome: selected clinical issues. PMID- 3041608 TI - The effect of surfactant replacement therapy on conditions associated with respiratory distress syndrome: patent ductus arteriosus, intraventricular hemorrhage, and bronchopulmonary dysplasia. AB - In assessing the current experience with SRT, both considering its effects on respiratory function and on associated problems, there are strong suggestions that this therapy has the potential to have a major impact on the morbidity and mortality of premature infants. The studies to date have not disclosed any serious adverse reactions, and there are strong trends toward improvements in short-term respiratory function and in long-term outcomes. In particular, SRT does not seem to adversely effect cardiopulmonary circulatory status and worsen the effects of the patent ductus arteriosus. There are no adverse effects on intraventricular hemorrhage and some possibility that it may lead to a reduction in hemorrhages. The use of SRT will likely have its biggest effect on reducing the incidence and severity of bronchopulmonary dysplasia. PMID- 3041609 TI - The use of C-reactive protein in the diagnosis of renal allograft rejection. PMID- 3041610 TI - Role of renal ultrasonography (RUS) and micturating cystourethrogram (MCU) in the assessment of vesico-ureteric reflux (VUR) in children and infants with urinary tract infection (UTI). PMID- 3041611 TI - Nasal CPAP therapy in the obstructive sleep apnoea syndrome. PMID- 3041613 TI - Manganese exposure and biological indicators. PMID- 3041612 TI - Fosfomycin. PMID- 3041615 TI - [40th anniversary of the World Health Organization]. PMID- 3041614 TI - Tuberculosis in patients undergoing dialysis--a reappraisal. PMID- 3041616 TI - [Organization of a referral service for urban populations (scientific review)]. PMID- 3041617 TI - [An important stage of cooperation in the field of medicine]. PMID- 3041618 TI - [The Bacteriological Institute of the Kharkhov Medical Society as a base for the training of medical personnel]. PMID- 3041619 TI - [From the history of the control of alcoholism]. PMID- 3041620 TI - Colony hybridization to identify mammalian cells containing amplified, transfected, or expressed sequences. AB - The identification of variant cell clones has played an important role in the elucidation of various biochemical pathways. Such clones are typically selected by altering the medium or the growth conditions to give a selective advantage to the desired variant. The availability of cloned DNA probes has made it possible to identify bacterial colonies or viral plaques containing desired sequences, even in the absence of selection. We describe here a new method for the identification and isolation of mammalian cells containing any sequence for which a probe exists. Mammalian cell clones are first replicated onto nylon cloth. The original clones and the replicas are grown for a few days to enlarge the clones. All the clones on the original dish are transferred in situ to a filter and processed for hybridization. By altering the processing, either DNA or RNA sequences in the clones can be identified. When a clone of interest is located by hybridization, the cells can be isolated from the nylon replicas. We expect this method to be of value in the isolation of clones containing amplified, transfected, or mutated genes, as well as those exhibiting altered gene expression. PMID- 3041621 TI - Isolation and characterization of Chinese hamster cell line resistant to monofunctional alkylating agents. AB - A clonal derivative of a Chinese hamster Don D-6 cell line resistant to methyl methane sulfonate (MMS) has been isolated following mutagenesis by ethyl methane sulfonate (EMS). The clone, designated as MMSr-1, exhibited high resistance to killing by the monofunctional alkylating agents MMS and EMS. This characteristic had not been acquired by a transient adaptation to the alkylating agents but was found to be a stable heritable trait. MMSr-1 was more sensitive to high-molecular weight chemicals, such as colchicine and puromycin, than Don D-6. Both MMSr-1 and its parental cells showed the same ability to take up radioactive MMS. The resistance of MMSr-1 appears not to be due to altered uptake of MMS. The resistance was accompanied by low chromosomal aberration and sister chromatid exchange (SCE) induction but not by mutability. Protein synthesis inhibitors such as cycloheximide and puromycin reduced the resistance to the same level as that in Don D-6. SCE induction by MMS in this clone was not antagonized by the protein synthesis inhibitors, whereas mutagenesis was reversed to the normal parental cell level by these inhibitors. Aphidicolin, a DNA-synthesis inhibitor, exhibited no such effects. These results suggest that MMSr-1 might have modified repair capacity, which can be normalized by treatment with the protein-synthesis inhibitors, for lethal DNA damage by monofunctional alkylating agents, and that SCE formation by the alkylating agents is closely correlated with chromosomal aberration and cell lethality. PMID- 3041623 TI - Improving conventional insulin regimens. PMID- 3041622 TI - Sequence specificity of mutations induced by benzo[a]pyrene-7,8-diol-9,10-epoxide at endogenous aprt gene in CHO cells. AB - We have determined the spectrum of mutations induced by +/--trans-7,8-dihydroxy 9,10-epoxy-7,8,9,10-tetrahydrobenzo[a] pyrene (BPDE) at the endogenous aprt locus in an hemizigous Chinese hamster ovary cell line exposed to 0.7 microM BPDE. Southern analysis of 59 independent mutants revealed no major genomic alterations, indicating that gene inactivation was the result of a point mutation. This conclusion was confirmed by the cloning and sequencing of 21 of these mutants. The predominant mutation, the G:CT----T:A transversion, comprised 62% of the spectrum, but other base pair substitutions and frameshifts were recovered. An examination of the target sequences for BPDE mutation revealed that mutations were localized within runs of G:C base pairs. However, approximately half of these G:C runs involved a particular sequence--a run of guanines flanked by adenine residues. Of seven such sites within the coding sequence of aprt, mutations were clustered within five of them. This class of sequence occurs at codon 61 of the human C-Ha-ras 1 protooncogene and may account for the selective activation of this codon by BPDE. PMID- 3041625 TI - The relevance of left ventricular bands. AB - Left ventricular bands (LVBs) were first described almost a century ago but were largely ignored until their 'rediscovery' by echocardiography. Renewed interest in LVBs and the demonstration of their association with clinical abnormalities have resulted in attempts to establish their causal relationship with various phenomena, especially the vibratory systolic murmur and premature ventricular contractions. Published articles on LVBs are reviewed with specific reference to nomenclature, embryological development, histological features, prevalence, demonstration in vivo and at autopsy, and alleged clinical importance. Current views on LVBs are evaluated and future research directions are suggested. PMID- 3041624 TI - Techniques for the diagnosis of malaria. AB - Malaria is endemic in large parts of the world and is subject to control by health authorities. The microscopic examination of many thousands of blood films for parasites, an essential component of surveillance and control schemes, is a tedious and time-consuming task. In recent years research has been directed at alternative methods to improve the mass diagnosis of malaria. These include immunoassays for the detection of antigen and the use of genetic probes. The techniques at present available are reviewed and some of the merits and disadvantages of the various tests discussed. PMID- 3041626 TI - Primary arteritis of the aorta and its branches. AB - Primary arteritis of the aorta and its branches is a single clinicopathological entity affecting one or more segments of the aorta and resulting in a variety of symptom complexes. The prevalence is not known, but it is an important cause of renovascular hypertension in the black population of South Africa. Recently published articles on primary arteritis of the aorta and its branches are reviewed, and an attempt is made to draw conclusions regarding its cause, clinical features and treatment. PMID- 3041628 TI - The AIDS test. PMID- 3041627 TI - Intertrochanteric fractures in an elderly white South African population. AB - Intertrochanteric fractures of the hip are a common problem in any elderly population; 168 patients over 50 years of age were reviewed 13 or more months after surgery. The male to female ratio was 1:5. The 12-month mortality rate was 30.95% and slightly less than 50% of patients walked normally. Prognostic factors which may assist in patient management are identified. PMID- 3041629 TI - A comparison of an enzyme immunoassay and cell culture for detection of Chlamydia trachomatis in genito-urinary specimens. AB - Chlamydia trachomatis infection is easy to treat but laborious to diagnose by culture. An antigen-detecting enzyme immunoassay (EIA; Chlamydiazyme, Abbott Laboratories, North Chicago, IL), suitable for testing many samples, was compared with the conventional iodine-stained, one-passage culture. A total of 471 duplicate samples from 218 men (urethra) and 128 women (urethra and cervix) attending a sexually transmitted disease clinic were examined by both tests. The overall sensitivity, specificity, and predictive value of positive and negative results were 80.3%, 96.7%, 86.4%, and 95.0%, respectively. No difference between male and female patients was observed. A remarkable difference between sensitivities of female urethral (60.0%) and cervical (95.5%) samples was found. This difference is clearly important in cases of women with urethral infection only (19% in our study) and points to the need for further improvement of this EIA. PMID- 3041631 TI - Detection of IgG and IgA antibodies to Chlamydia trachomatis in sera of patients with chlamydial infections: use of immunoblotting and immunoperoxidase assays. AB - The immune response to individual structural polypeptides of Chlamydia trachomatis was studied in 75 sera from symptomatic and asymptomatic women with culture-proved genital infections and from apparently healthy women who were culture-negative for C. trachomatis. The immunoblotting technique and the single serovar (L2) inclusion immunoperoxidase assays were used for measurement of the various antibodies. Antibodies to 18 structural polypeptides, ranging in molecular weight from 29 to 204 Kdaltons, were detected by the immunoblotting technique in sera from seropositive women. The immunoperoxidase assay showed that sera with high titers of IgG and IgA antibodies to C. trachomatis reacted with more polypeptides than did sera with low titers in this test. Antibodies to the 60- and 62-Kdalton polypeptides were detected in almost all sera positive for IgG and IgA antibodies, irrespective of chlamydial shedding. About 40% of sera with high IgG and IgA titers reacted with 39-, 57-, 64-, 72-, 86-, 105-, 155-, and 204 Kdalton polypeptides. The prevalence of IgA antibodies to C. trachomatis was higher among women with culture-proved chlamydial infections than among apparently healthy controls. PMID- 3041632 TI - [Apropos of adoption. Paris region]. PMID- 3041630 TI - Comparison of cefpimizole with cefotaxime and penicillin G for treatment of uncomplicated gonorrhea. AB - One hundred sixteen patients (92 men and 24 women) with suspected uncomplicated gonorrhea were randomized in a double-blind manner to receive intramuscular treatment with 1.0 g of cefpimizole, 1.0 g of cefotaxime, or 4.8 x 10(6) units of aqueous procaine penicillin G (APPG) with 1 g of oral probenecid. Seventeen percent were nonassessable (cultures negative, co-existing syphilis, etc.). Infection sites in 96 assessable patients were urethra (78), cervix (17), pharynx (two), and rectum (two). Of 52 patients treated with cefpimizole, 46 (88%) were bacteriologically cured, as compared with 100% (24 of 24) treated with APPG (P = 0.18) and 90% (18 of 20) treated with cefotaxime (P greater than 0.20). On a weight basis the in-vitro activity of cefpimizole against Neisseria gonorrhoeae was similar to that of APPG. Pain at the injection site was reported by 52% of patients treated with cefpimizole as compared with 27% of those given cefotaxime (P = 0.008) and 17% of those given APPG (P = 0.002). No major organ toxicity was found with cefpimizole, cefotaxime, or APPG. Thus, for acute uncomplicated gonorrhea cefpimizole is similar in efficacy to cefotaxime and APPG but has a higher frequency of pain at the injection site. PMID- 3041633 TI - The importance of arteriographic interpretation in occlusion or pseudo-occlusion of the carotid artery. AB - Carotid arteriography can be misleading in that roentgenographic occlusion of the internal carotid artery (ICA) may be suggested when the artery is actually anatomically patent. The distinction of occlusion versus pseudo-occlusion is crucial in recommending proper treatment. Aroused by the misinterpretation of two arteriograms by our radiology departments, a review of 780 arteriograms done on 780 patients during a three year period was begun. Of these, eight (1.0 per cent) symptomatic patients had conflicting arteriographic reports, with the radiologists reporting complete occlusion and the authors describing pseudo occlusion. Established arteriographic criteria for pseudo-occlusion of the ICA are emphasized. These eight patients underwent carotid arterial exploration and all were found to have patent ICA. After successful carotid endarterectomy, seven of the eight patients have remained asymptomatic and have had no hemodynamically significant stenosis on noninvasive testing in the follow-up period, ranging from two to 38 months. Therefore, in evaluating patients with carotid territory symptoms, knowledge of the established arteriographic criteria for ICA pseudo occlusion should alert the physician to the possibility of ICA patency. If this is suspected, the patient should undergo exploration of the carotid artery with subsequent endarterectomy if patency is demonstrated. PMID- 3041634 TI - Primary tuberculous enteritis. AB - Because of the large influx of immigrants from the Third World into this country, primary enteric tuberculosis, although rare, can still be found in all sections of the United States. The fact that it may mimic the more common neoplastic and granulomatous diseases, coupled with the relative paucity of instances, increases the likelihood of a misdiagnosis. Diagnosis may be further complicated by a false negative Mantoux test (purified protein derivative skin test) and lack of evidence of tuberculosis elsewhere in the patient. Adequate treatment consists of appropriate chemotherapeutic agents and the judicious use of surgical resection. Given appropriate treatment, the outcome in most patients should be favorable. PMID- 3041635 TI - Role of intraoperative fibrinolytic therapy in acute arterial occlusion. AB - Nineteen patients with acute onset of ischemia affecting the lower extremities were studied from January 1985 to March 1987. Patients with preoperative Doppler and angiographic studies consistent with arterial occlusions subsequently underwent a thromboembolectomy using a Fogarty catheter. All patients were given a bolus injection of 5,000 units of heparin intravenously at the start of the surgical procedure. In all patients studied, a clot was retrieved on the first pass, but after two additional passes, total distal blood flow was not shown to be restored on angiogram. Intraoperative angiograms showed distal emboli. All patients underwent intraoperative fibrinolytic therapy by local bolus infusion. Streptokinase, ranging from 50,000 to 200,000 units, was administered in 50,000 unit injections in ten to 15 minute intervals. Repeat attempts at thromboembolectomy with the Fogarty catheter resulted in an additional clot retrieved in all 19 patients with intraoperative angiographic, Doppler and clinical improvement. No perioperative or postoperative complications were observed, including anaphylactic reactions, uncontrollable bleeding or amputation. Four patients had nonacute femoropopliteal bypass operations within the next six months. Intraoperative fibrinolytic therapy can be a safe and effective adjunct in acute arterial embolic occlusion requiring balloon catheter thromboembolectomy. PMID- 3041636 TI - Endotoxin and pulmonary cell injury. AB - The physiopathologic similarity between adult respiratory distress syndrome (ARDS) secondary to sepsis and endotoxin-induced pulmonary abnormalities has provided extensive descriptive information confirming bacterial endotoxin as a factor initiating the heterogeneous pulmonary changes in ARDS. The present studies have used an established in vitro model for pulmonary cell injury to examine bacterial endotoxin 1, as a direct cytotoxic agent on the two major alveolar cell types, pulmonary endothelium and epithelium; 2, as a stimulant of neutrophil-mediated pulmonary cell injury, and 3, to examine effector mechanisms of cell-mediated damage by studying the potential effectiveness of antioxidants and antiproteolytic agents in the inhibition of this process. Endotoxin direct toxicity and stimulation of neutrophil-mediated pulmonary cell injury was observed in both pulmonary cell populations in systems free of activated serum complement. Endothelial cells were observed to be more susceptible to both the direct effect of endotoxin and to neutrophil-mediated injury when compared with epithelial cell derived monolayers. The addition of an antiprotease (soybean trypsin inhibitor [STI]) was superior to antioxidants (catalase, superoxide dismutase) in reducing the neutrophil-mediated endothelial toxicity (stimulated 51CR per cent release) observed. A 92 per cent degree of protection was observed with the highest dose of STI (5 milligrams per milliliter) used. Proteases released by activated neutrophils on endotoxin stimulation appear to be the predominant toxic species responsible for endothelial injury in this system. PMID- 3041638 TI - Orbital surgery: repair of the frontal fossa by "en bloc" removal and self replacement of the orbital roof. Technical note. AB - The orbital roof may be repaired with implants or duplication of the frontal bone flap. A simple and safe method is presented in which a flap of orbital roof is "en bloc" removed and replaced to reconstruct the normal bony anatomy of the frontal fossa. PMID- 3041637 TI - Giant peripheral aneurysm of the posterior inferior cerebellar artery treated with excision and end-to-end anastomosis. AB - A patient with a giant aneurysm arising from the tonsillomedullary segment of the posterior inferior cerebellar artery (PICA) presented with clinical and computed tomography findings suggestive of spontaneous cerebellar hemorrhage. Magnetic resonance imaging led to arteriography and the correct diagnosis. Lack of a clippable neck on the aneurysm and its location proximal to the choroidal point prompted treatment by excision of the aneurysm and end-to-end anastomosis of the PICA. No neurological deficit resulted from the procedure. PMID- 3041639 TI - [The role of postoperative radiotherapy in the treatment of hypernephroid carcinoma]. AB - The following results can be derived from the clinical and radiobiological literature as well as from the authors's own experiences with 91 patients analyzed retrospectively and evaluated with respect to their risk factors. 1. A postoperative irradiation is very probably sensible in patients with tumor perforation through the renal capsule (T3/T4) and with venous or lymph node manifestations. 2. The operation method is essential, above all the transabdominal technique and the quality of lymph dissection. 3. Prospective studies are necessary in order to clear up the role of radiotherapy in patients with lymph node manifestations and the other two risk factors mentioned in item 1. 4. Further studies on the tumor biology of the hypernephroid carcinoma are necessary in order to gain prognostic criteria allowing a pretherapeutic sensitivity recognition. 5. The effect of a supplementary radiotherapy can be only that of local recurrence prevention in case of a locally advanced tumor and the absence of demonstrable remote metastases. This seems justified, because the local recurrence rate in our own patients was only 3.3%. PMID- 3041640 TI - Reversal of abnormal glucose production after pancreatic resection by pancreatic polypeptide administration in man. AB - Pancreatic polypeptide (PP) deficiency has been associated with impaired hepatic sensitivity to insulin and pancreatogenic diabetes in chronic pancreatitis. Since pancreatic resection might also result in PP deficiency, hepatic responses to insulin infusion (0.25 mU/kg/min) were determined by the euglycemic glucose clamp technique in 10 patients who had previously undergone pancreatic resection for trauma and in eight healthy control subjects. Six resection patients (RES-PP) demonstrated deficient PP levels, with a mean increase of plasma immunoreactive PP of 20 +/- 7 pg/ml above basal rate after a test meal compared with 232 +/- 82 pg/ml in control subjects (p less than 0.01) and 353 +/- 133 pg/ml in four other patients undergoing resection with normal levels of immunoreactive PP (RES + PP) (p less than 0.03). Three identical insulin infusion studies were performed in each subject, the second of which was performed during the final 2 hours of an 8 hour infusion of bovine PP (2.0 pmol/kg/min). Whereas hepatic glucose production (HGP) in control subjects fell 74% +/- 4% from a basal rate of 2.0 +/- 0.1 mg/kg/min to a 60- to 120-minute value of 0.5 +/- 0.1 mg/kg/min during insulin infusion, HGP was suppressed only 58% +/- 5% in RES-PP subjects, from 1.9 +/- 0.1 to 0.8 +/- 0.1 mg/kg/min (p less than 0.05 vs controls). Intravenous infusion of PP corrected the hepatic resistance to insulin seen in the PP-deficient group. During PP infusion, HGP was suppressed 74% +/- 5% in RES-PP subjects, from 2.1 +/ 0.2 to 0.5 +/- 0.1 mg/kg/min (p less than 0.04 compared with initial study). PP infusion produced no significant change in glucose metabolism in control and RES + PP subjects. Overall glucose disposal rates were not altered by PP infusion in any group. These findings support a role of PP as a glucoregulatory hormone and suggest that PP deficiency may serve as a reversible pathophysiologic factor in the abnormal glucose metabolism seen after pancreatic resection. PMID- 3041641 TI - Inhibition of tumor high-energy phosphate metabolism by insulin combined with rhodamine 123. AB - The purpose of this study was to determine whether the energy metabolism of an experimental rodent sarcoma was selectively depressed by the combination of inhibition of glycolysis and respiration. In vivo phosphorus-31 nuclear magnetic resonance spectroscopy was used to monitor the response of tumor or brain high energy phosphate compounds to insulin hypoglycemia, rhodamine 123, or both agents in fasting rats with subcutaneous methylcholanthrene-induced sarcomas. Insulin or rhodamine 123 alone produced a similar 50% to 60% reduction in tumor adenosine triphosphate (ATP) concentration compared with controls injected with saline solution (p less than 0.05, one-way analysis of variance [ANOVA]). The combination of insulin plus rhodamine 123 resulted in a 90% reduction of tumor ATP concentration, which was significantly different from the effect of either agent alone (p less than 0.05, one-way ANOVA). Brain phosphocreatine and ATP concentrations were unchanged by these agents. Administration of dimethyl sulfoxide (DMSO)/glycerol, the vehicle for rhodamine, produced a 35% reduction of tumor ATP, which was similar to the effect of insulin alone but significantly different from rhodamine. The combination of DMSO/glycerol plus insulin hypoglycemia resulted in a 70% reduction in tumor ATP, which was significantly elevated compared with the combination of rhodamine plus insulin. Glucose deprivation induced by insulin, and combined with the inhibition of oxidative phosphorylation, produces an additive depression of tumor energetics. The drug vehicle DMSO/glycerol significantly depresses tumor energy metabolism, presumably because of its DMSO component, which may explain the previously reported antineoplastic efficacy of this solvent. Combinations of inhibitors directed at different points of tumor metabolism produced an enhanced depression of tumor energetics, whereas host tissue was protected. PMID- 3041643 TI - Small-bowel bacterial overgrowth and systemic immunosuppression in experimental peritonitis. AB - The effect of intraperitoneal infection on small-bowel flora and on systemic immunity was studied in a rat model, with use of the delayed-type hypersensitivity (DTH) response to keyhole limpet hemocyanin (KLH) as a measure of global immunologic integrity. Twenty-four hours after the induction of peritonitis by cecal ligation and puncture, concentrations of Escherichia coli in the proximal gastrointestinal tract increased from fewer than 10(3) colony forming units (CFU)/ml to more than 10(9) CFU/ml, and the DTH response decreased from 10.0 +/- 0.2 to 2.1 +/- 0.4 mm. To assess the contribution of this altered luminal flora to the observed suppression of DTH scores, cecal ligation without puncture was performed in a group of animals whose endogenous flora had been suppressed by administration of oral neomycin. Oral administration of live antibiotic-resistant E. coli to the study animals resulted in significant DTH depression compared with controls given saline solution (2.7 +/- 0.4 vs 4.4 +/- 0.4 mm, p less than 0.005), even though the gastrointestinal tract was anatomically intact. Similar depression was seen if the challenge was limited to the small bowel as a result of the prior performance of an ileostomy and occurred in the absence of significant systemic or portal levels of viable bacteria. The results suggest that gut endotoxin plays a role in the immunosuppression associated with peritonitis. PMID- 3041642 TI - Fatty acid intake and Kupffer cell function: fish oil alters eicosanoid and monokine production to endotoxin stimulation. AB - Diets high in n-3 fatty acids appear to have an anti-inflammatory effect, which is thought to be due to decreased macrophage prostaglandin (PG) and thromboxane (Tx) production after incorporation of these fatty acids into cell membrane phospholipids. The effect of n-3 fatty acids incorporation on macrophage monokine release in response to septic stimuli is not well established. Kupffer cells, the fixed macrophages of the liver, were obtained from rats fed diets with fat sources derived from corn oil (CO, control), fish oil (FO, high in n-3 fatty acids), or safflower oil (SO, high in n-6 fatty acids) for 2 or 6 weeks. After exposure to bacterial lipopolysaccharide, Kupffer cells from rats fed FO for 2 or 6 weeks produced less PG and Tx than Kupffer cells from rats fed CO or SO. After 2 weeks of defined diets, interleukin-1 (IL-1) and tumor necrosis factor release were not affected by dietary fat source. In contrast, after 6 weeks of feeding, Kupffer cells from both the FO and the SO groups released less IL-1 and tumor necrosis factor when triggered by lipopolysaccharide than Kupffer's cells from animals fed the control diet that contained CO. These data suggest that altered monokine release from macrophages may contribute to the anti-inflammatory effect of diets high in n-3 fatty acids. Also shown in our results is that prolonged changes in membrane phospholipid content induced by dietary fat source can influence not only PG and Tx production but monokine release as well. PMID- 3041644 TI - Pancreas transplantation in nonuremic, type I diabetic recipients. AB - Between July 1978 and January 1988, 111 of 210 pancreas transplants were in nonuremic, nonkidney (NUNK) recipients in whom complications of diabetes were judged more serious than the potential side effects of antirejection therapy. In all NUNK cases, the 1-year patient survival rate (PSR) and graft survival rate (GSR) were 90% and 39%. Since November 1984, 1-year PSR and GSR for 62 NUNK recipients of pancreas transplants alone (PTA) were 93% and 48%, compared to 89% and 37% for 28 pancreas transplants after (PAK) and 89% and 73% for 20 simultaneous (SPK) with a kidney transplant. The 1-year GSR for 1984-88 technically successful (TS) PTA cases (n = 47) was 63%, versus 75% for PAK (n = 13) and 86% for SPK (n = 17) cases. The 1-year GSRs, by technique and source for 1984-88 PTA cases, were 58% for bladder-drained cadaver (n = 30), 51% for enteric drained related (n = 32), and 29% for enteric-drained cadaver (n = 17) donor transplants, and 75% (n = 24), 77% (n = 16), and 38% (n = 13) for the corresponding TS cases. Of NUNK recipients, 35 have had grafts function for more than 1 year and all but 3 were treated with cyclosporine (CsA). In the CsA treated recipients, serum creatinine increased and creatinine clearances decreased by a magnitude of 40% during the first 6 months but thereafter remained stable in all but 2 patients. In the patients with functioning grafts studied at 1 year, retinopathy remained stable in 59% and progressed in 41% of the eyes (n = 39). Motor nerve conduction velocities were significantly increased and muscle action potentials remained stable in 24 patients with functioning grafts studied at 1 year, while in 14 patients in whom transplants failed there was a significant decrease in evoked muscle action potentials. A sustained euglycemic, insulin-independent state can be established in nonuremic, nonkidney diabetic recipients of pancreas transplants alone, with a beneficial effect on neuropathy, lack of an immediate benefit on advanced retinopathy, and an effect on nephropathy compounded by the influence of CsA on renal function. PMID- 3041645 TI - Adherence of granulocytes to nylon fibers. Evidence for a plasma granulocyte adherence factor. PMID- 3041647 TI - A tribute to Robert Maxwell founder and publisher of Pergamon Press. PMID- 3041646 TI - Determination of the enzymatic activity of brinase and of the brinase inhibitor capacity by the azocollagen assay. PMID- 3041648 TI - Cellular Ca2+ homeostasis and Ca2+-mediated cell processes as critical targets for toxicant action: conceptual and methodological pitfalls. AB - Because of the central role of the calcium messenger system in diverse functions of tissues, organs, and cells, Ca2+ homeostasis and function may prove to be critical cellular and molecular targets for a diverse range of toxicants. Experimental proof of these targets as a specific site of toxicant action is challenging and technically difficult as a result of the complexity of Ca2+ homeostatic and Ca2+-mediated processes. However, as the investigation of normal physiological control of Ca2+ and function will continue to be an active and productive area of basic research for several years to come, it is anticipated that these insights will be increasingly applied to the understanding of the mechanisms of action of toxic agents. PMID- 3041650 TI - Anticoagulant therapy in cerebrovascular disease: review and meta-analysis. AB - Sixteen acceptably randomized studies of anticoagulant therapy after cerebral or retinal ischemia or infarction are reviewed and the results among 1,046 anticoagulated patients and 1,071 controls are analyzed. The following conclusions are derived. 1) Anticoagulant therapy has not been shown to be better than control management after transient ischemia or nonprogressing ischemic stroke; this is true whether the control management was deliberately ineffectual treatment (generally studies completed in 1974 or earlier) or platelet antiaggregant therapy (pooled results of three recent studies). 2) Although a study done 30 years ago demonstrated no benefit, a recent study showed benefit from anticoagulant therapy in patients who had had cerebral emboli of cardiac origin; additional controlled data are needed. 3) There is evidence that patients with thrombosis in evolution might benefit from anticoagulant therapy; additional controlled data are needed. PMID- 3041649 TI - Transcranial Doppler ultrasonography of carotid-basilar collateral circulation in subclavian steal. AB - The combination of a carotid-basilar and a vertebro-vertebral collateral circulation was verified directly in a patient with a complete subclavian steal by means of transcranial Doppler ultrasonography. The patient showed permanently reversed blood flow in the basilar artery. The subclavian steal influenced the hemodynamics of the circle of Willis at rest and during functional tests of the collaterally supplied arm. Our investigation provides the first direct experimental evidence of increased blood flow velocity in the carotid artery after decompressing the collaterally supplied arm. PMID- 3041651 TI - Blood glucose and stroke. PMID- 3041652 TI - Atrial fibrillation and stroke: new ideas, persisting dilemmas. PMID- 3041653 TI - Clinical and hemodynamic significance of innominate artery lesions evaluated by ultrasonography and digital angiography. AB - To determine the hemodynamic and clinical consequences of an atherosclerotic obstructive lesion of the innominate artery on the cerebral circulation, 20 patients with an innominate artery lesion underwent neurologic examination and ultrasonic duplex scanning before and after right arm exercise. The patients were divided into two groups: Group 1, 12 patients with 40-80% stenosis and Group 2, eight patients with 80-100% stenosis. A significant difference between the groups was noted in both the hemodynamic and clinical manifestations. All 12 Group 1 patients compensated for the increased demand for blood of the right arm through the innominate artery itself, and only one showed symptoms of vertebrobasilar insufficiency associated with right arm exercise. In all eight Group 2 patients, compensation through the innominate artery failed; six (75%) showed symptoms of vertebrobasilar insufficiency after exercise. Dynamic duplex scanning is well suited to investigate stenotic lesions of the innominate artery, the effects of arm exercise on the development of cerebral symptoms, and the source of blood flow to the arm. Dynamic duplex scanning proved to be useful in selecting patients who may be candidates for direct arterial surgery. PMID- 3041654 TI - Duplex scanner study of carotid artery dissection following surgical treatment of aortic dissection type A. AB - In patients suffering from aortic dissection, persistent perfusion of the false lumen distal to the implanted graft is frequent. Postoperative follow-up examinations of the carotid arteries of these patients were performed by duplex scanner and correlated with clinical symptoms. Thirty-nine patients who survived the surgical treatment of acute type A aortic dissection had duplex sonography of both common carotid arteries after an average postoperative follow-up of 53 months. In 21 cases a composite graft and in 18 cases a supracoronary prosthetic vascular graft were implanted. No sign of residual dissection of the common carotid arteries was seen in 23 patients; in nine there was a dissection of both common carotid arteries, and seven patients had a unilateral carotid dissection (five right, two left). There were nine symptomatic patients with the following symptoms: transient ischemic attack (four), amaurosis fugax (four), stroke with incomplete recovery (one). Two symptomatic patients had a corresponding dissection. The generally good prognosis of all these patients suggests a conservative nonoperative treatment. PMID- 3041655 TI - Use of the caprinised strain of rinderpest virus for potency testing an attenuated cell culture rinderpest vaccine. AB - The caprinised strain of rinderpest virus was inoculated into goats to produce a challenge stock. These goats were kept with control animals (goats, sheep, calves). In this trial the caprinised strain was shown to have a mild pathogenicity for goats and it spread to one of two contact goats but not from goats to other species. The caprinised strain was then tested on cattle where a febrile reaction was observed. The caprinised strain also did not spread between cattle. The cattle vaccinated with a freeze-dried vaccine produced from the attenuated Kabete RBKO strain on bovine kidney cells were then challenged with the caprinised strain with good results. PMID- 3041656 TI - Prevalence of piroplasmosis in equines in the Colombian province of Cordoba. AB - Eighty-two equine sera from 13 farms in northern Colombia were examined for antibodies to Babesia caballi and B. equi using the complement fixation (CF) and the indirect fluorescent antibody (IFA) test. Seroreactors to both piroplasms were present on all farms. The IFA test indicated a prevalence of 90% for B. caballi and 94% for B. equi. The CF test detected antibodies to B. caballi in 41% and to B. equi in 65% of the animals. The prevalence of seroreactors in different age groups revealed a significant decline in CF antibodies to B. caballi in animals older than three years. IFA titres for both Babesia spp. gradually declined with increasing age of the animals but were still present in most animals of the oldest age group (over nine years old). Anocentor nitens was found on all farms whereas Amblyomma cajennese was found only on two farms located on the coast. PMID- 3041657 TI - Immunohistochemical study of ten cases of argyrophilic carcinoma (carcinoid) of the breast. AB - The significance of argyrophilia in human breast cancer is still a controversial issue. We tested immunohistochemically 10 cases of argyrophilic carcinomas of the breast and found evidence of immunoreactivity with neuroendocrine markers: chromogranin, NSE, gastrin, insulin and bombesin. Argyrophilia was demonstrated in breast cancers of the usual types and was found to be related to the secretory activity of neoplastic cells. Unfortunately, no adequate follow-up data are available to clarify the natural history of argyrophilic breast cancer. A clinical treatment different from that of conventional breast cancer is not at present justified. PMID- 3041658 TI - Fine aspiration versus fine cutting needle, and comparison between smear cytology, inclusion cytology and microhistology in abdominal lesions. AB - Two hundred patients underwent ultrasound guided percutaneous fine needle biopsy of focal solid abdominal lesions using 22 gauge aspiration and cutting needles. The material obtained by aspiration needle was treated by smear cytology and by inclusion cytology, and that obtained by cutting needle by microhistology. The retrieval rate was 89% for aspiration needle (smear cytology = 89%, inclusion cytology = 83.5%) and was 83% for cutting needle; the combined diagnostic accuracy was 98%. The typing accuracy was 76% for smear cytology, and was 84% for inclusion cytology and microhistology. We conclude that: 1. to obtain the highest retrieval rate (98%) both aspiration and cutting needles are necessary, because the aspiration needle is more likely to secure necrotic or soft tissue, and the cutting needle, fibrous or hard tissue; 2. histologic treatment of the samples yields a higher typing accuracy: 84% vs 76%; however, smear cytology remains essential because it permits a much faster evaluation of the adequacy of the sample and because it may avoid histologic methods in 76% of cases; 3. the smear cytology + microhistology combination seems to be the best solution (retrieval rate = 97.5%), but the costs are much higher because the cutting needle is somewhat expensive. The best solution would be to use the combination smear + inclusion cytology (retrieval rate = 89%) and to reserve the cutting needle for when aspiration needle material proves to be inadequate. PMID- 3041659 TI - Comparative study of two immunocytochemical techniques in the electron microscopical detection of alpha-1-antitrypsin in routinely processed liver biopsies. AB - Two indirect immunocytochemical techniques using different markers, namely peroxidase and gold, were applied to ultrathin sections of liver biopsies to detect alpha-1-antitrypsin. The blocks used were taken from the routine electron microscopy files and had been processed optimally for maximum ultrastructural preservation. The immunogold technique provided the best method for localizing alpha-1-antitrypsin and was associated with ultrastructural preservation equivalent to that seen in routinely processed liver biopsies. The procedure may be a useful adjunct to understanding the pathogenesis of alpha-1-antitrypsin deficiency and in recognizing further variants of this disorder. PMID- 3041660 TI - [Malignant hemopathies in adults and bone marrow graft]. PMID- 3041661 TI - [Allogeneic bone marrow transplantation and myeloid leukemia in the chronic and accelerated stages]. PMID- 3041663 TI - Renal allotransplantation using cadaveric or living related donors: long-term results. AB - During the period of 1968 through 1975 renal allografts were performed using cadaveric or living related donors. At that time the typing as it is known today was unavailable. Despite this serious limitation 8 long-term survivors living from eight to more than fourteen years postrenal allotransplantation are presented. All had normal renal function and are without detectable serious side effects of long-term immunosuppression. PMID- 3041662 TI - [The periodic medical examination]. PMID- 3041664 TI - Adolescent males who insert genitourinary foreign bodies: is psychiatric referral required? AB - Although much is known about the urologic aspects of genitourinary foreign bodies, the psychiatric profile of the adolescent male who inserts a foreign body into his urethra is not available. Six cases from a urologic service are presented, and the pertinent literature is reviewed. Psychiatric consultation for such selfinserters will allow the development of such a profile. An appropriate management plan can then be formulated. PMID- 3041665 TI - Bilateral ureteral obstruction secondary to endometriosis. AB - Endometriotic ureteral obstruction is an uncommon but serious event often unrecognized until hydronephrotic renal atrophy has occurred. A case of bilateral endometriotic ureteral obstruction treated with danazol (Danocrine) is reported, and the literature is reviewed. PMID- 3041666 TI - Primary localized amyloidosis of ureter. AB - Primary localized amyloidosis of the ureter is very rare, and only 22 cases have been reported in the world literature. We report the twenty-third case along with a review of the relevant literature. Due to its radiologic resemblance to malignancy, many cases were treated by nephroureterectomy in the past. The case being reported here was successfully treated by local excision and ureteroneocystostomy. PMID- 3041667 TI - Correlation of radionuclide and ultrasound studies with biopsy findings for diagnosis of renal transplant rejection. AB - The diagnosis of early renal transplant rejection is of the utmost importance to the transplant recipient. Unfortunately, such a diagnosis is often extremely difficult to make. In an attempt to clarify this issue we retrospectively evaluated 35 patients with the presenting diagnosis of rejection for the correlation of comparable radionuclide (RN) and ultrasound (US) examinations with biopsy findings. In 21 patients with heavy interstitial mononuclear cell infiltration, 22 of 23 serial RN studies within forty-eight hours of biopsy were positive for rejection. Only 3 of 14 comparable US studies were positive for rejection. When examinations performed within approximately fourteen days were evaluated, 7 of 11 RN studies were positive for rejection, while 2 of 9 comparable US studies were positive for rejection. However, in 14 patients with mild or no interstitial cellular infiltration, only 6 of 13 RN studies were positive, while all 4 US examinations were negative. In the group evaluated at approximately two weeks, 2 of 6 RN studies were positive, while 0 of 5 US studies were positive. We conclude that the serial RN study is more sensitive than US examination for the diagnosis of acute rejection. US, however, proved valuable in the identification of transplant complications (i.e., fluid collections, ascites, and hydronephrosis). PMID- 3041668 TI - Evaluation of erectile impotence. PMID- 3041669 TI - [Immunoenzyme, turbidimetric and affinity chromatographic methods of determining the fibronectin level in human plasma (comparability of results and effect of conditions and duration of blood storage)]. AB - Immunoenzymatic, turbidimetric and affinity chromatography methods used for estimation of fibronectin concentration exhibited the similar results but the turbidimetric procedure was more simple and less time-consuming. Concentration of fibronectin was not decreased in blood maintained in polymer container "Gemacon 500" within 48 hrs both at 4 degrees and 20 degrees, while about 40 = 50% of fibronectin was lost after storage of blood in glass flasks under similar conditions. PMID- 3041670 TI - [Regulation of the activity of the oxoglutarate dehydrogenase complex of animal origin (review)]. AB - Recent data on regulation of the multi-enzyme oxoglutarate dehydrogenase complex from pigeon breast muscle, porcine heart, bovine adrenal glands and kidney are reviewed. The most characteristic property of the complex consists in activation of the trigger oxoglutarate dehydrogenase component by and ATP. Action of these agents is more pronounced at low concentrations of 2-oxoglutarate and is directed towards alteration of the substrate half-saturation value SO.5. The adrenal oxoglutarate dehydrogenase complex exhibits positive cooperativity of allosteric ADP-binding sites which improved its sensitivity to variations in the effector concentration. The oxoglutarate dehydrogenase complex appears to be not only an important functional component but also is a regulatory unit of the Krebs cycle. PMID- 3041671 TI - [Age-related characteristics of insulin regulation of the physico-chemical properties of liver membranes]. AB - Alterations in physico-chemical properties of rat liver plasmatic membranes induced by insulin were shown to be age-dependent The experiments showed also that high doses of the hormone impaired distinctly the membranes in old animals. The relationship between structural alterations in plasmatic membranes and the rate of lipid peroxidation was observed. PMID- 3041673 TI - [The biorhythms of renal activity in healthy persons (review of the literature)]. PMID- 3041672 TI - [Structural aspects of the transport function of serum albumin (review)]. PMID- 3041674 TI - [Side effects of drugs in elderly persons as an indication of the changes in pharmacokinetics and pharmacodynamics]. PMID- 3041675 TI - [Echography in the diagnosis of diseases of the thyroid gland]. PMID- 3041676 TI - [The use of etaden in patients with peptic ulcer]. PMID- 3041677 TI - [The mechanism of the action of trental]. PMID- 3041678 TI - [Treatment of patients with pulmonary hypertension caused by chronic nonspecific lung diseases (review of the literature)]. PMID- 3041680 TI - [The weakness of individual psychologic dream theory]. AB - This article undertakes a critical evaluation of Adlerian dream theory. The main weakness of the theory is found to be its lack of an inherent instance of truth that shows the dreamer the way to a better and more feasible life style. Contemporary Adlerians' treatment of the master's dream dogmas and their practical use in psychotherapy are described. There seems to be a convergence movement of today's practical application methods of the dream in all psychotherapeutic schools. Adlerian dream interpretation in the original sense intended by Adler is practised nowhere by psychotherapists today and seems largely antiquated. PMID- 3041679 TI - [Experiences with kidney transplantation in elderly patients]. AB - Despite a significant increase in the number of elderly patients with end stage renal disease, these patients still represent a minority of renal transplant recipients in many countries. Roughly the recipients in the present study were older than 50 years of age. Infection was a more common complication in these patients than in the younger patients. However, the incidence of cerebrovascular and cardiovascular complications was found to be lower than expected. Patient graft survival in 110 renal transplants in 106 patients aged 50 years or older were 87% and 76% respectively. These results suggest that cadaveric renal transplantation represents a relatively safe form of therapy also for older patients. PMID- 3041683 TI - Aminopyrine metabolism in primary monolayer cultures of diabetic rat hepatocytes. AB - 1. A new support system has been used which provides long-term maintenance of rat liver parenchymal cells in monolayer cultures. The cells, maintained on collagen gel/polychlorinated vinylidene film, expressed aminopyrine metabolizing activity for up to 5 days. This culture system was utilized to study the metabolism of aminopyrine in the liver cells isolated from normal, alloxan- and streptozotocin diabetic rats. 2. Aminopyrine was metabolized at a slower rate in both types of cultured diabetic rat hepatocytes than in cultured normal rat hepatocytes, as judged from higher levels of the unchanged drug in the culture medium. 3. The formation of the metabolites 4-monoaminoantipyrine, 4-acetylaminoantipyrine and 4 formylaminoantipyrine decreased in the cultured diabetic rat hepatocytes, while that of 4-aminoantipyrine was at the same levels as controls. In contrast, 3 hydroxymethyl-2-methyl-4-dimethylamino-1-phenyl-3-pyrazolin-5-on e (AM-CH2OH) formation in the cultured diabetic rat hepatocytes increased over control value. These findings agree with in vivo results which have been reported by the authors. 4. The increase in AM-CH2OH was prevented by insulin in a dose-dependent manner. However, insulin did not affect the formation of other metabolites. These findings indicate that the amount of cytochrome P-450 isozyme involved in the oxidation of 3-methyl group may be regulated by insulin. 5. The present results, indicate that this primary culture system is a useful tool for the study of drug metabolism in diabetes. PMID- 3041684 TI - [Possibilities and trends of pharmacotherapy of cardiac arrhythmias in childhood and adolescence. 1: General principles, clinical aspects and indications]. PMID- 3041685 TI - [Overtime work and ischemic heart disease]. PMID- 3041682 TI - Cefotaxime i.v. versus oral neomycin-erythromycin for prophylaxis of infections after colorectal operations. PMID- 3041681 TI - Microsurgical free fibular bone transfer: a technique for reconstruction of large skeletal defects following resection of high-grade malignant tumors. PMID- 3041686 TI - [Passive smoking in controversy]. PMID- 3041687 TI - [First evidence of the biological effect of radium rays--a further contribution of Otto Walkhoff]. PMID- 3041688 TI - [Peculiarities of theophylline therapy in childhood]. AB - Profound knowledge of the pharmacokinetics of theophylline is a prerequisite to successful therapy. In children the clearance of theophylline is considerably greater than in adults. The required higher dosage in relation to body weight results in more pronounced oscillations of drug levels in serum. Investigations in 280 children in the age of 1 to 18 years with bronchial asthma, treated with theophylline compounds, confirm the necessity of individual dosage and surveillance of the drug level in serum. Instead of the determination of the drug level in serum, its measurement in saliva is sufficiently reliable and less molesting. Checks of drug level are also a valuable educational measure to improve patients' compliance with drug intake. The effectivity of asthma therapy in children could be decisively improved by an adequate formulation of a slow release preparation of theophylline. PMID- 3041689 TI - [Lung function findings in workers in the rubber industry]. AB - Chronic obstructive lung disease is common in workers in the rubber industry. Because of the close relation between hyperreactivity of the airways (HR) and the annual decline in FEV1 we measured HR in 89 healthy workers of a big rubber plant. Three groups were formed (control, latex production, rubber production). The prevalence of HR was that of a healthy general population (PC 20 FEV1 less than 0.2% Methacholin in 3 cases). In the lower Methacholin range a separation between all groups was possible. PMID- 3041690 TI - [Initial status of the medical management of pediatric and adolescent type I diabetic patients]. AB - Based upon a large number of type I diabetics, representative of diabetes in childhood and adolescence of the GDR and taking into consideration communication in the literature, the present position is commented on autoimmune pathogenesis, immune intervention, frequency of complications, optimizing of therapy and on social-medical aspects of the diabetic child in school. Besides first degree diabetes heredity, degree of HLA identity with the index case, disturbances of glucose tolerance and insulin secretion, auto-immunological phenomena directed against islet cells have to be considered without doubt as risk factors for diabetes. But absolutely reliable markers of an imminent diabetes onset do not exist. The frequency of somatic retardations and diabetes-specific complications is low under multiple insulin injection regime, used in 87 per cent. However, their occurrence alone should stimulate to remove existing differences between clinical achievable metabolic compensation (good: 60%; bad: 2.5%) and longterm metabolic control (good: 40%; bad: 22%). In order to realize this aim, medical care is orientated mainly on qualification of patients and their parents to self adaption of the insulin dose regime in dependence on values of home-monitoring. PMID- 3041691 TI - [Russia-Halle interaction in medicine of the 18th century. I: The medical organizers Johann Deodat Blumentrost (1676-1756) and Laurentius Blumentrost (1692 1755)]. AB - In the Russian medicinal and scientific organization of the early 18th century decisively promoted by Czar Peter I such personages as the physicians Johann Deodat Blumentrost and Laurentius Blumentrost have obtained important key positions. Last not least their work was characterized by inspirations which they received at the university of Halle the destination of many of their compartriots in the Petrine and Postpetrine era. At the Petersburg Academy of Sciences which began its work in 1725 Laurentius Blumentrost was its first president. PMID- 3041692 TI - [The renal artery and uterine circulation in normal and toxemic pregnancies]. AB - The renal and uterine arteries of 52 patients with normal course of pregnancy and 12 patients with EPH gestosis were examined by Doppler sonography between the 35th and 38th weeks of gestation post menstruationem. Doppler curves of renal arteries were recorded in a further 31 nonpregnant subjects. A significant difference in arterial vessel resistance was found between patients with normal course of pregnancy and those with EPH gestosis. There was also a significant difference in the circulation parameters in the renal vessels between patients with normal courses of pregnancy and the nonpregnant controls. However, there were no differences in this vessel segment between the patients with EPH gestosis and the nonpregnant controls. Among patients with EPH gestosis the renal increase in resistance was relatively more pronounced than the uterine increase. In addition to the calculated flow parameters there were also marked differences in the Doppler curves of these two groups. There is evidently a considerable increase in renal circulation in normal pregnancies. This is in agreement with findings described in the literature, obtained by invasive methods. On the other hand, a clear limitation of renal and uterine resistance was seen in the patients with EPH gestosis. This ties in with the well-known morphologic changes in the kidneys in cases of EPH gestosis. A larger patient sample will have to be studied to determine whether the renal vascular changes described here may represent an early sign of EPH gestosis. If this is confirmed it would make sense to include Doppler sonography of the renal vessels in routine diagnostic procedure. PMID- 3041694 TI - [Sampling of the chorionic villi: transcervical fine-needle and endoscopic aspiration]. AB - In 63 cases chorionic villi sampling has been performed under complete anesthesia just before legal abortion; nine of them were endoscopic transcervical and 54 transcervical by means of a catheter under direct ultrasonic control. 81% offered useful chorionic villi, 17.4% only decidual material. The last 21 aspirations by means of a catheter between the 8th and the 12th week of gestation (p.m.) caused no problem at all. From the first punction on useful chorionic material has been obtained. PMID- 3041693 TI - [Improved diagnosis of extrauterine pregnancy by endosonography]. AB - Between July 1987 and January 1988, 44 patients with a tentative diagnosis of ectopic pregnancy underwent sonographic examination by means of vaginal probe at the Department of Gynecology and Obstetrics of the RWTH Aachen. The sonographic findings, all of which were confirmed by subsequent clinical and or surgical clarification, were as follows: an ectopic pregnancy was diganosed in 16 cases, an early intrauterine pregnancy in seven, an intrauterine abortion in seven, and in one case a uterine malformation-a dermoid cyst and a functional cyst. In 11 cases sonographic examination showed the interior genital region to be normal, with no sign of pregnancy. In the 16 ectopic pregnancies diagnosed, it was possible in 13 cases to visualize the pregnancy directly by sonography, including the amniotic sac, and to make measurements. In one case a normally developed ectopic pregnancy with living embryo was seen at the end of the seventh week of gestation post menstruationem. In the remaining three cases the diagnosis was established on the basis of an empty cavum uteri associated with a slightly enlarged uterus and demonstration of fluid in the pouch of Douglas. In two cases the ectopic pregnancy was correctly localized by "feeling" with the intravaginal probe to establish the cause of circumscribed pain. In three case the tentative diagnosis of an ectopic pregnancy made on the basis of sonographic findings was not confirmed by subsequent clarification procedures. The results described show that in most cases ectopic pregnancies can be demonstrated directly by sonography using an intravaginal probe.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3041695 TI - [Ultrasound echographic imaging and measurement of the amniotic cavity and yolk sac in early pregnancy: comparative study of intact and disordered pregnancies]. AB - The chorionic cavity, vertex-breech length, yolk sac, and amnion were systematically demonstrated and measured in 50 early pregnancies, i.e., between the end of the sixth and the tenth week of gestation post menstruationem (p.m.). This was done in 23 cases with clinically and sonographically intact pregnancies without symptoms of abortion, in 12 cases with living embryos with symptoms of abortion, and in 15 cases of retained miscarriage. The thin amniotic membrane is sonographically demonstrated as a narrow, sharply defined reflected band in the chorionic cavity. In addition to direct demonstration of the amniotic membrane, the amniotic cavity can be demonstrated by a density difference in the echogenicity of the chorionic and amniotic fluids. While there are delicate, homogenously distributed inner echoes in the chorionic cavity, the amniotic cavity is empty save for the embryo structure. Sonographically, therefore, the amniotic cavity stands out as a spherical structure within the chorionic cavity. In all pregnancies with a living embryo it was possible to demonstrate the yolk sac sonographically as a sharply defined ring structure in the chorionic cavity. In 10 of the 15 cases of retained miscarriage only a rudimentary remnant of the yolk sac could be detected. The development of the amniotic and chorionic cavities and vertex-breech length was constant in the pregnancies with living embryos, the amnion developing synchronously with the vertex-breech length.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3041697 TI - [Ultrasonic diagnosis of congenital uterine abnormalities]. AB - 1-2% of women has abnormal uterine development due to nonunification of the Mullerian ducts in the embryonal period. At the RWTH Aachen, in the department of gynaecology and obstetrics, between January and June 1987, we had searched systematically for maldevelopment of the uterus in 2299 echosonografies. In 13 cases we found maldevelopment of internal genital; 5 of these cases were diagnosed by an echosonografic routine-examination. The echografic criteria of the different grades of uterine malformations have been determined, systematized and discussed in relation to the symptoms. The most frequent malformations as uterus subseptus, uterus septus, uterus bicornis and uterus duplex are subject of a detailed discussion. This work demonstrates that echosonografic is a very efficient instrument to diagnose uterine malformations and gives us a very exact anatomic interpretation of malformations. PMID- 3041696 TI - [Non-invasive diagnosis of pre- and postpartum urination disorders]. AB - Postpartal disorders of urinary discharge due to birth trauma are frequently the cause of rising infections of the urinary tract during the puerperium. In diagnosis, noninvasive methods should be preferred. Bladder-emptying function was checked by means of uroflowmetry and sonographic measurement of residual urine in 83 patients at the Department of Gynecology and Obstetrics at the RWTH, Aachen, in the third trimester and post partum. A considerable restriction of urine flow was found both prepartally as well as on the third day post partum. In contrast, there was practically no disturbance of bladder emptying on the sixth day post partum. This function is restored more quickly in cases of cesarean section than in cases of vaginal delivery. As early as the third day post partum no further influence of subpartal analgesia could be demonstrated. The findings failed to show any difference between the group with vaginal/surgical delivery and that with spontaneous birth. Generous use of these noninvasive diagnostic methods to identify postpartal micturition disorders can be recommended. PMID- 3041699 TI - [The suprachiasmatic nucleus of the hypothalamus as a regulator of the circadian system of mammals]. PMID- 3041698 TI - [Organization of the motor representation of the facial musculature in the neocortex of various orders of mammal]. PMID- 3041700 TI - [Heat and mass exchange of the lungs with the environment]. PMID- 3041701 TI - Coping with OBRA. PMID- 3041702 TI - Remembering Leonard Weyl. PMID- 3041703 TI - [Various problems of the development of the theory of evolution in the light of the integral function of a dialectic materialism method]. PMID- 3041704 TI - [The organization of higher cortical functions in man in various forms of reinforcement]. PMID- 3041705 TI - [Bases for the selection of individuals for relatively labile physiological traits exemplified by body heat resistance]. PMID- 3041706 TI - [Effect of immunization with Escherichia coli J5 on the course of experimental pyelonephritis in rats]. AB - Among the germs of urinary tract infections is a great heterogenicity. Therefore, in search of vaccine candidates of pyelonephritis such germs must be checked the antigens of which to be occurred as much in other pyelonephritis germs as possible. E. coli J5 as a re-mutante of E. coli O111 has a core-antigen which is found in the most enterobacteria. We have examined whether a pyelonephritis in rats was prevented by prophylactic use of a E. coli J5 vaccine and if the course of the disease was influenced by vaccine treatment of animals with pyelonephritis, respectively. Neither a pyelonephritis was prevented nor an existing infection was influenced favourable by the vaccine of E. coli J5. PMID- 3041707 TI - [Chronic circulatory disorders of the digestive organs of the abdominal cavity. Causes, clinical aspects and treatment]. AB - Chronic arterial insufficiency of blood supply to digestive organs in the abdominal region is a specific manifestation of circulatory diseases. It is rare but may cause substantial danger to the patient. Reference is made in this paper to international literature as well as to cases involved in vascular surgery at the Berlin School of Medicine (Charite) for an account of present knowledge on aetiology, pathogenesis, symptoms, diagnosis, and therapy of chronic disorders of abdominal blood supply. Techniques of vascular repair represent an appropriate therapeutic concept, accurate diagnosis and critical indication provided. PMID- 3041709 TI - [Indications and diagnostic criteria for early ventilation of trauma patients]. AB - Mechanical ventilation is the potential therapeutic approach to traumatic brain lesion and acute adult respiratory distress syndrome (ARDS) in the wake of severe injury in an accident. Aggravating cerebral symptoms, such as non-targeted defence reactions in coma, hemiplegia, synergism of extension, convulsions, pontine respiratory disorders, and intracerebral pressure beyond 30 Torr are diagnostic criteria for immediate mechanical ventilation of patients with brain trauma. The same action is indicated for cases of ARDS exhibiting, on top of the typical constellation of causes, hypoxia below 60 Torr paO2 and vital capacity below 15 ml/kg body weight or respiratory rates in excess of 30/min. PMID- 3041708 TI - [Initial clinical experiences with heart transplantation in East Germany]. AB - An account is given in this paper of first experience from orthotopic heart transplantation. Six of eight patients with transplants have so far survived, with two of these having been in very good general condition for more than a year. All surgical indications were derived from myocardial dilatation. A triple combination of cyclosporine A, prednisone, and azathioprine was used for immunosuppression. PMID- 3041711 TI - [Clinical experiences in the use of "autogenized" spongiosa]. AB - Autogenic spongiosa is of the highest biological valence. The principle of autogenisation of allogenic spongiosa is described in this paper. Reference is made to quantitative and qualitative advantages of the procedure. PMID- 3041710 TI - [Treatment of unstable thoracolumbar spinal injuries--determination of its present status and a preliminary report on transpedicular stabilization]. AB - Follow-up checks were made on 19 patients with instabile injuries of the thoracolumbar spine. Poor late results were recorded from eleven of them who had undergone conservative or inadequate surgical treatment. Axial malposition of severe clinical disorders were among the findings. Transpedicular osteosynthesis of instable fractures of thoracic or lumbar regions of the spine has been practised by the authors since 1986, with plate fixation of only two vertebral segments. Pathoneurological disorders are handled by laminectomy or hemilaminectomy followed by H-shaped interspine bone grafting. Posterior rim fragments, burst off the spine, are either reduced or removed. Intraoperative perimyelography is possible. Therapeutic exercises are initiated immediately after the operation. After wound healing, the patient is allowed to get out of bed, supported by a plaster corset. Eleven patients have so far undergone the above surgical treatment. Four of them have resumed their original jobs, whereas three patients with paraplegia have been transferred to a rehabilitation centre. PMID- 3041713 TI - [Sexuality in the aged]. AB - Sexuality in older age group persons, important for human self reliance, is often underestimated or ignored. But the possibility to be orgastic can maintain up to the postmenopausal lifetime of a woman, depending on a fulfilled partnership. Somatic troubles, connected with the estrogen deficiency-syndrome, can be treated with oral or local estrogens. PMID- 3041712 TI - [Ligament replacement-plasty in chronic ruptures of the collateral ligament of the finger joints using the ligamentum retinaculare obliquum]. PMID- 3041715 TI - Changes in indices of ventricular filling, size and function during graded levels of lower body negative pressure: comparison between normal and transplanted hearts. PMID- 3041714 TI - Echocardiographic assessment of the effect of therapy on left ventricular function in congestive heart failure. PMID- 3041717 TI - [Royal Belgian Society of Surgery. List of members]. PMID- 3041716 TI - Evaluation of captopril effect in congestive cardiomyopathy by a computerized noninvasive method. PMID- 3041718 TI - Neopterin: an useful biochemical marker in the monitoring of allogeneic bone marrow transplantation. PMID- 3041719 TI - Endoscopic brushing cytology and biopsy in the diagnosis of upper gastrointestinal tract lesions. A study of 350 cases. AB - Direct-vision endoscopic examination conducted on 4,000 patients for persistent upper gastrointestinal (GI) complaints over a period of five years revealed 350 visible lesions that were subjected to brushing cytology and biopsy. Cytologic examination of brushing smears from all 350 cases showed malignant cells in 67 (19.14%), cells suggesting benign polypoid neoplasms in 4 (1.14%), ulcerative and reparative features with attendant atypias in 186 (53.14%), inflammatory findings in 91 (26%) and false-negative findings in 2 cases (0.57%). Only 259 (74%) of the visible lesions were also subjected to endoscopic biopsy. Of the 67 patients with positive cytology, 52 were judged positive on the biopsy specimen; the 2 false negative cytologic reports were confirmed as positive by biopsy. In four patients with gastric ulcers, malignant cells were seen along with gastric repair cells. This study indicates that brushing cytology is very useful in detecting benign ulcerative lesions with their atypias, a feature that could be useful in monitoring and controlling lesions in high-risk groups of patients, such as in India. In this study, endoscopic brushing cytology gave a better diagnostic yield than did endoscopic tissue biopsy. However, the two techniques are complementary for the diagnosis of upper GI malignancies. PMID- 3041720 TI - The role of brushing cytology in the diagnosis of gastric malignancy. AB - The results of endoscopic biopsy and brushing cytology in 234 consecutive patients with established histologic diagnoses of discrete gastric lesions were analyzed. A histopathologic diagnosis of malignancy, established by independent means, was made in 74 patients. Brushing cytology was positive for malignancy in 63, a diagnostic sensitivity of 85%. Endoscopic biopsy was positive in 64, a diagnostic sensitivity of 86%. The sensitivity for combined cytology and biopsy was 91%, which was not significantly greater than for biopsy alone (P = .6). Cytology yielded false-positive results in 5 of 160 patients (3.1%) with confirmed benign disease. There were no false-positive biopsy reports. Although both brushing cytology and biopsy have high diagnostic sensitivities, based on the findings of this study, the routine addition of cytology to biopsy in the endoscopic evaluation of gastric lesions is not recommended. Cytology could be reserved for situations in which difficulty is encountered in obtaining adequate tissue for histologic examination and for cases with a high suspicion of malignancy that have yielded negative biopsies. PMID- 3041721 TI - Malignant ascites of serous papillary ovarian adenocarcinoma. An immunocytochemical study of the tumor cells. AB - In 17 malignant peritoneal effusions due to papillary serous adenocarcinoma of the ovary, the reaction patterns of the tumor cells to monoclonal antibodies (MAbs) against surface antigens were studied and compared with the reaction patterns of mesothelial cells in the same effusions. The following surface markers were used with the adhesive slide method: epithelial membrane antigen (EMA), human epithelium-specific cell surface antigen (HEA-125), human endothelial antigen (BMA-120), carcinoembryonic antigen (CEA 3-13), an antibody against natural killer cells and cytotoxic cells (BMA-070), granulocyte antigen (Leu M1) and leukocyte antigen of class I (HLA-1). In all cases, from 30% to 95% of the tumor cells reacted with EMA and HEA-125. Tumor cells showed a positive staining with CEA 3-13 in only five cases. In all cases, from 75% to 95% of the tumor cells reacted positively with BMA-120. The reactivity of a few mesothelial cells with EMA and of all mesothelial cells with BMA-120 did not interfere with the identification of positive tumor cells since the reaction patterns were different. Interestingly, our study demonstrated that BMA-070, an MAb identifying natural killer cells and cytotoxic cells, is also a most useful tumor marker. The same was found to be true for Leu M1, an MAb originally thought to react only with granulocytes. The tumor cells showed a partial or total loss of the expression of HLA-1 reactivity. Since all cases were immunocytochemically positive for tumor cells while conventional cytology was positive in only 13 of the cases, the immunocytochemical analysis of malignant peritoneal effusions due to papillary serous adenocarcinoma of the ovary seems able to improve the cytologic diagnosis of the fluids. PMID- 3041722 TI - Liesegang-like rings in fine needle aspirates of renal/perirenal hemorrhagic cysts. AB - Periodic structures with equally spaced radial striations, identified as Liesegang-like rings, were encountered in fine needle aspirates of two patients' hemorrhagic renal/perirenal cysts. The patients, one 60 and the other 39 years old, had acute right-flank pain; both underwent nephrectomy. The ring structures ranged in size from 8 microns to 200 microns in diameter and had regularly striated double walls. Histochemical and immunoperoxidase tests for iron, calcium, mucopolysaccharides, amyloid, keratin and hemoglobin performed on the tissue sections of the resected specimens gave negative results. With electron microscopy, the ring structures of one of the cases displayed a fine fibrillary composition. Since these Liesegang-like structures may be mistaken for parasites or necrotic tissue, pathologists should be aware of them. To our knowledge, this is the first report of Liesegang-like rings in the cytology literature. PMID- 3041723 TI - Primary malignant pleural tumors (mesotheliomas) presenting as localized masses. Fine needle aspiration cytologic findings, clinical and radiologic features and review of the literature. AB - The cytologic and histologic appearances of four localized primary malignant pleural tumors occurring in three patients subjected to fine needle aspiration are presented. In one case, the radiographic mass was the only manifestation of what was found to be a diffuse epithelial mesothelioma at surgery. Two other patients had localized primary sarcomatous mesotheliomas, of low grade and high grade, respectively. In the latter patient, a second metachronous but identical tumor appeared on the contralateral side during follow-up. The clinical findings, radiologic features and cytologic differential diagnosis of both the epithelial and the sarcomatous variants are discussed, together with a review of the literature pertinent to localized primary malignant pleural tumors. PMID- 3041724 TI - Diagnosis of metastatic adamantinoma of the tibia by pulmonary brushing cytology. AB - A case of adamantinoma of the tibia metastatic to the lung is reported in which the metastatic lesion was initially diagnosed by pulmonary brushing cytology. The cytologic features, including clusters of small cells with either prominent nucleoli or spindle-shaped hyperchromatic nuclei, appear to be distinctive enough to differentiate this lesion from other metastatic malignant tumors of the lung. PMID- 3041725 TI - A simple method for the preparation of paraffin-embedded cell blocks from fine needle aspirates, effusions and brushings. AB - A simple method for the preparation of paraffin-embedded cell blocks from cytologic specimens obtained by fine needle aspiration, by brushing or from effusions is described. The cells are fixed in suspension in 50% ethanol for one hour and pelleted by centrifugation in a 50-mL plastic tube. The fixative is removed, and the pellet is suspended in 3 mL of acetone for dehydration for ten minutes and thereafter repelleted. The acetone is then removed, and the cell pellet is dried at 60 degrees C for one hour. Melted paraffin is added onto the dry warmed cell mass and allowed to solidify at room temperature. A conical paraffin block with the cells in the top is obtained and can be handled as a routine tissue block. PMID- 3041726 TI - Value of the filter imprint technique in the cytologic study of ocular lesions. PMID- 3041727 TI - Comparative plasma thyroglobulin measurements with three non-cross-reactive monoclonal antibodies in metastatic thyroid cancer patients. AB - In 12 normal individuals and 25 patients with metastases of differentiated thyroid cancer, plasma thyroglobulin (Tg) concentrations were measured simultaneously with three immunoradiometric assays. Each of the three systems used a different, non-cross-reactive monoclonal Tg antibody as second antibody. In general, there was a very close correlation between the results of the three different systems in the cross-sectional study of the 25 cancer patients as well as in longitudinal follow-up studies in selected patients. The monoclonal antibody Tg 40 produced values about 30% higher than the two other systems. The difference was, however, not statistically significant owing to the large scatter. The monoclonal antibody Tg 13 appeared to be very sensitive to interference with thyroglobulin autoantibodies. In conclusion, the monoclonal antibodies against the three epitopes tested produced very similar Tg values in normal individuals and 25 patients with metastatic thyroid cancer; however, before more is known about epitope specificity of Tg autoantibodies and heterogeneity of tumour Tg, monoclonal antibodies should be used for routine measurements only with caution. PMID- 3041728 TI - Influence of growth factors on an insulin-producing cell line (RINm5F). AB - The influence of IGF-I, insulin and epidermal growth factor (EGF) as well as glucose as control was studied on both growth and function of RINm5F cells, an insulin-producing cell line derived from rat insulinoma tissue. [3H]thymidine incorporation and DNA content (fluorometrically) were measured for estimating cell growth, insulin release and biosynthesis [( 3H]leucine incorporation) as parameters for cell function. There was a significant increase in both [3H] thymidine incorporation and DNA content into RIMm5F cells by glucose already at very low concentrations. IGF-I and high concentrations of insulin too were able to stimulate cell growth (insulin also in additional experiments with isolated rat islets). EGF, however, was without growth effect on this cell line (in contrast to previous own results in isolated islets). Insulin secretion and biosynthesis were also increased by very low glucose concentrations. IGF-I and EGF were ineffective regarding these functional parameters in RINm5F cells. Besides for glucose, our results demonstrate a role for IGF-I and insulin, but not for EGF, in regulating growth of these cells. PMID- 3041730 TI - Current management of traumatic chylothorax. AB - The development of a traumatic chylothorax is an uncommon but serious clinical entity. Two cases of traumatic chylothorax are reported. The anatomy and physiology of the thoracic duct and the etiology, diagnosis and management of traumatic chylothorax are discussed. PMID- 3041729 TI - Further evidence that preoptic anterior hypothalamic GABAergic neurons are part of the GnRH pulse and surge generator. AB - The in vivo release rates of GABA from the preoptic/anterior hypothalamic area (PO/AH) of ovariectomized rats were assessed by means of a focal perfusion cannula system. Seven days after surgery all animals received a sc silastic capsule implant containing either estradiol-17 beta (E2) or corn oil, and they were perfused 3 days later. Perfusate fractions were sampled at 5-min intervals and blood was collected every 10 min over a period of 5 h. In ovariectomized animals PO/AH GABA release was pulsatile without any diurnal rhythm. Prior to frequency analysis by means of the pulsar-programme, LH and GABA values were z transformed. Significant LH peaks of all examined ovariectomized rats were superimposed and GABA data were arranged accordingly. It became evident that LH episodes are preceded by a significant reduction of preoptic anterior hypothalamic GABA release. The secretion patterns of GABA and LH were profoundly affected by E2 replacement. During early noon when LH levels were low, constantly elevated hypothalamic GABA release rates were observed in E2-substituted rats in comparison to ovariectomized rats. GABA release rates fell significantly during the E2-induced LH surge. Our previous demonstration of the existence of a large number of estrogen-respective GABAergic neurons in the PO/AH is suggestive of these neurons changing their activity in response to estrogen treatment. We conclude that these estrogen-respective GABAergic neurons are involved in the generation of GnRH pulses as well as in the generation of the so-called positive feedback effect of E2 on LH release.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3041731 TI - The De Hemptinne inhaler. AB - The De Hemptinne ether inhaler was presented to the Royal Academy of Medicine of Belgium in February, 1847 by Auguste De Hemptinne, a pharmacist. This article outlines the details of construction and use of this device. A comparison to contemporary inhalers is also made. PMID- 3041733 TI - Carl-Axel Hamberger 1908-1988. PMID- 3041732 TI - Hemodilution and autotransfusion. A blood sparing and safety program. PMID- 3041734 TI - Cochlear morphology from Wurzburg (1951) to Turin (1987): old and new aspects. AB - A review is given of the methods employed in human temporal bone pathology, from the camera lucida drawings of the last century to the sophisticated techniques of today. PMID- 3041735 TI - Immunohistochemical investigation of the human inner ear. Limitations and prospects. AB - The application of highly sensitive and specific immunohistochemical methods in routine light microscopy has been rewarded by a great many new observations. The essential underlying antigen-antibody reaction has allowed the recognition and identification of some unknown or unidentified components of the cell. Successful results are best achieved by means of reliable fixation and the use of frozen sections. Frozen sections of the human cochlea are, however, impaired by unavoidable damage to the tissues to be studied. The temporal bone has to be decalcified, resulting in a reduction of the range of suitable methods and an interference with a reliable interpretation of the results. The preservation of the specific antigenicity of the tissues is of paramount importance, allowing the detection of the investigated antigen by the antibody applied. The investigation of different fixatives and various methods of decalcification in tissues possessing some familiar and readily identifiable antigens has confirmed the immunohistochemical suitability of properly fixed and adequately decalcified human temporal bones. The present demonstration deals with some of the principal technical procedures and includes examples of their application for the study of the human inner ear. The goal of our research is the development of reliable immunohistochemical methods of fixation and decalcification to be employed in the study of specimens from patients with Meniere's disease, sudden deafness, progressive loss of hearing, genetic sensorineural syndromes, etc. This will lead to the extension of our knowledge of some unrecognized causes of sensorineural hearing loss. PMID- 3041736 TI - Light and electron microscopical changes seen in acute tubular necrosis in renal allografts. AB - Seven renal biopsy specimens taken from three renal-allografted patients clinically suspected of having acute tubular necrosis were examined by light microscopy and five of these specimens were also examined by electron microscopy. The common findings in these three patients were as follows: The tubular lumina in all parts were dilated and the tubular epithelia were flat. Large vacuoles were occasionally observed within the tubular epithelial cell cytoplasm. These alterations were prominent in proximal convoluted tubules. Electron microscopically, the microvilli of the brush border of proximal convoluted tubules were loose and relatively short. The basal infoldings of proximal convoluted tubules were reduced or had disappeared. It could not be confirmed whether the large intracytoplasmic vacuoles apparent by light microscopic observation were intracytoplasmic or widened lateral intercellular spaces upon electron microscopy. In the most markedly damaged allograft of the three, a grayish hematoxylinophilic substance, which corresponded to autophagosomes with electron-dense round bodies upon electron microscopy, was often observed in the tubular epithelia. In addition, immature or regenerative tubular epithelia were observed. Most of these alterations were similar to those of acute ischemic change seen in non-renal transplantation. PMID- 3041737 TI - [Resolution of enantiomers and diastereoisomers by HPLC and its in pharmaceutica analysis]. PMID- 3041739 TI - Microbial ecology of the terrestrial subsurface. AB - We have presented a current view of the microbial ecology of the terrestrial subsurface by considering primarily the ecology of shallow aquifer sediments. The properties of the aquifer sediments and groundwater determine their ability to support microbial life and control the abundance and activities of microorganisms. Pore size, nutrient limitations, availability of electron acceptors, and large surface area for attachment all may have major effects on microbial abundance and activities in aquifer material. Microorganisms are the predominant forms of life in the subsurface. They will be found wherever enough space, nutrients, and water are available for them to live. Environmental factors such as pH, temperature, hydrostatic pressure, and dissolved salts also may influence subsurface microbial populations, but these factors do not exhibit great extremes in shallow water table aquifers, and thus only in very deep formations might they limit diversity or preclude the existence of microorganisms. Although the presence and activity of microorganisms in most subsurface environments are predictable, only recently have subsurface microbial populations in shallow subsurface zones been characterized. Aseptic sampling methods have been employed and microbiological and biochemical methods have been adapted to determine the types, abundance, and metabolic activities of microorganisms in subsurface material. Bacteria dominate, but eukaryotic microorganisms also are present. Vertical profile studies of a shallow aquifer in Oklahoma showed that active microbial biomass declined with depth to the unsaturated zone, but was variable in saturated sediments. Such a distribution of active biomass may be common in shallow aquifers. Studies on the lateral distribution of microorganisms in shallow and deep aquifers suggest that microorganisms are transported or migrate over fairly long distances in aquifer sediments. Surficial aquifers may be colonized by vertical or lateral transport and migration of surface microorganisms from recharge areas, but microorganisms could also have colonized when sediments were originally deposited. The biological and physical mechanisms controlling the migration of microorganisms in aquifers are not well understood. The function of shallow aquifers was considered with regard to nutritional ecology. Most pristine aquifers are oligotrophic. Heterotrophic life in these unique ecosystems is supported by secondary organic compounds that filter down from the soil above. The quantity and quality of organic nutrients depend on the age of water and rate of recharge of the aquifer.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3041738 TI - Activity and stability of recombinant human superoxide dismutase in buffer solutions and hypothermic perfusates. AB - The stability of recombinant human superoxide dismutase (r-hSOD) in buffer solutions was studied in solutions at various pH and temperatures. Additionally, we studied the effects of incubation with proteases, serum and two types of hypothermic perfusates. R-hSOD was stable in the pH range of 6-11 and at temperatures up to 80 degrees C for 30 min. R-hSOD activity was not affected by incubation with trypsin, aminopeptidase M or serum for 2 h. R-hSOD activity determined at various temperatures (4-37 degrees C) did not vary remarkably. R hSOD in hypothermic perfusates was stable at 4-37 degrees C for 24 h. PMID- 3041740 TI - Applications and mode of action of formaldehyde condensate biocides. PMID- 3041741 TI - Recovery of bioproducts in China: a general review. PMID- 3041742 TI - AIDS and HIV CNS disease: a neuropsychiatric disorder. PMID- 3041743 TI - Neuropsychological manifestations and predictors of HIV disease in vulnerable persons. PMID- 3041744 TI - Neuroimmunomodulation by opiates and other drugs of abuse: relationship to HIV infection and AIDS. PMID- 3041745 TI - Ethanol-associated immunosuppression. PMID- 3041746 TI - Psychoimmunology and AIDS. PMID- 3041747 TI - Immunomodulation by classical conditioning. PMID- 3041748 TI - Stress-associated immune suppression and acquired immune deficiency syndrome (AIDS). PMID- 3041750 TI - Behavioral risk factors and host vulnerability. PMID- 3041749 TI - Major life changes, chronic stress, and immunity. PMID- 3041751 TI - Legal and ethical issues in the neuropsychiatric research in AIDS. PMID- 3041752 TI - raf protooncogene expression in neural and immune tissues. PMID- 3041753 TI - Synthetic peptides in the study of the interaction of rabies virus and the acetylcholine receptor. AB - The neurotropism of some viruses may be explained in part by the attachment of these viruses to host cell receptors that are present on or even largely restricted to neurons. Rabies virus is an RNA virus that, after a period of replication in muscle, gains access to the central nervous system, where it selectively infects certain neuronal populations. The nicotinic acetylcholine receptor occurs in high density at the neuromuscular junction and is present in the central nervous system. Although several different cell surface constituents may act as attachment determinants for rabies, direct binding of radioactively labeled virus to affinity-purified acetylcholine receptor has been demonstrated. Binding of virus to the receptor was saturable and inhibited by up to 50% by alpha-bungarotoxin, a snake venom neurotoxin that binds at or near the acetylcholine binding site on the receptor. The molecular basis for the virus receptor interaction may lie in an amino acid sequence similarity between the snake venom neurotoxins and a segment of the rabies virus glycoprotein. Two peptides (10 and 13 residues) of the rabies virus glycoprotein and homologous bungarotoxin peptides were synthesized and tested for ability to compete with labeled alpha-bungarotoxin for binding to the acetylcholine receptor. The peptides were found to compete with toxin binding with affinities comparable to those of the cholinergic ligands d-tubocurarine and nicotine. These findings indicate that a segment of the rabies virus glycoprotein interacts with the acetylcholine receptor at or near the acetylcholine binding site of the receptor. The similarity between the virus glycoprotein and the neurotoxin was further evidenced by the cross reaction of antibody raised against the virus 10-mer with the bungarotoxin 10-mer. Binding of rabies virus to the acetylcholine receptor or to other neuronal bungarotoxin-binding proteins may be related to the neurotropism of this virus. In addition, knowledge of both the region of the virus involved in binding and the binding domain on the receptor may be helpful in developing new strategies for treatment, especially for viruses that infect the central nervous system or evade the immune response through genetic drift. These strategies include development of antiviral agents that cross the blood brain barrier and inhibit viral binding and the utilization as immunogens the regions of viruses, such as their binding domains, that are highly conserved among different strains. PMID- 3041754 TI - Voodoo death, the stress response, and AIDS. PMID- 3041755 TI - Concept and classification of dystonia. PMID- 3041756 TI - Clinicopathological correlation in symptomatic dystonia. PMID- 3041757 TI - Primary dystonias: a review of the pathology and suggestions for new directions of study. PMID- 3041758 TI - Investigation of dystonia. PMID- 3041759 TI - Hereditary progressive dystonia with marked diurnal fluctuation--consideration on its pathophysiology based on the characteristics of clinical and polysomnographical findings. PMID- 3041761 TI - Dopamine agonists and antagonists in the treatment of idiopathic dystonia. PMID- 3041760 TI - Dopa-responsive dystonia. AB - DRD is a distinctive clinical entity and an unexpectedly common subgroup of torsion dystonia. Diurnal fluctuation is often but not always present and does not reliably distinguish the disorder from ITD. DRD must be considered in the differential diagnosis of the child or adolescent presenting with a dystonic gait disorder, diplegic cerebral palsy, sporadic spastic paraplegia, ataxic syndromes, and juvenile parkinsonism. The response to L-DOPA is so dramatic and occurs so quickly that a diagnostic therapeutic trial should be undertaken in all patients presenting with these syndromes. PMID- 3041762 TI - Efficacy of dopamine agonists in dystonia. PMID- 3041763 TI - Blepharospasm and oromandibular-laryngeal-cervical dystonia: a controlled trial of botulinum A toxin therapy. PMID- 3041764 TI - An early description of dystonia: translation of Schwalbe's thesis and information on his life. PMID- 3041765 TI - Idiopathic torsion dystonia as described by Barraquer-Roviralta. PMID- 3041766 TI - Art of dystonia. PMID- 3041767 TI - Secondary dystonia. PMID- 3041768 TI - [Velopalatine disorders. An extensive review. 2]. PMID- 3041769 TI - Diagnosis of vasculogenic impotence. Comparing investigations by Doppler sonography and angiography. PMID- 3041770 TI - [Value of Doppler studies following intracavernous injection of papaverine hydrochloride in the diagnosis of impotence of arterial origin]. PMID- 3041772 TI - [Congenital curvatures of the penis]. PMID- 3041771 TI - Pudendal arteriography. PMID- 3041773 TI - [Priapism: personal approach to the problem in Kinshasa]. PMID- 3041775 TI - [A case of 2,8-dihydroxyadenine stones with a partial deficiency of adenine phosphoribosyltransferase]. AB - We report a case of 2, 8-dihydroxyadenine (2, 8-DHA) urolithiasis. A 39-year-old female was referred to our hospital with the complaint of right flank pain. An X ray examination showed right hydronephrosis. On May 1986, right percutaneous nephrolithotripsy was performed. Infrared spectroanalysis revealed 2, 8-DHA and calcium phosphate mixed calculus. The adenine phosphoribosyltransferase activity in erythrocytes was partially deficient. Since the operation, 300 mg/day of allopurinol has been administered, and there have been no signs of recurrence. PMID- 3041774 TI - [A case of renal malignant lymphoma]. AB - Malignant lymphoma affects any organ of the body, but is rarely found in a urological organ. We found a case of renal malignant lymphoma. A 45-year-old male, who had been operated on for primary hepatic malignant lymphoma 9 months previously, was admitted to our clinic, complaining of high fever. X-ray and ultrasonographic examinations suggested a metastatic tumor in the right kidney. Right nephrectomy was performed, and pathohistological examination revealed primary hepatic malignant lymphoma in the right kidney. PMID- 3041776 TI - [A case report of milk of calcium in a renal calyceal diverticulum]. AB - A 24-year-old male with epigastralgia was admitted to our Hospital because a kidney, ureter, bladder X-ray (KUB) revealed a right renal stone. The upright view revealed a crescent-shaped density with horizontal fluid level in the right kidney. With the patient supine the density was round. On retrograde pyelography the stone-like shadow was not enhanced. Since there were no symptoms nor evidence of renal disease other than the density within the right kidney, no treatment for the kidney was given. The patient is being followed in the urological clinic. PMID- 3041777 TI - [Hydronephrosis presenting as the first sign of malignant lymphoma of the small intestine: report of a case]. AB - A 45-year-old man was admitted to hospital on November 26, 1985 with the chief complaint of left hypochondrial pain. Excretory and retrograde pyelography revealed left hydronephrosis due to extrinsic obstruction of ureter. Computerized tomography and angiography revealed that a tumor of the small intestine was the cause of ureteral obstruction. In addition to the presence of a tumor, a fistula in the small intestine was disclosed on the upper gastrointestinal series. During the operation, a large mass which involved several organs was identified without mobility. The sophisticated operation was composed of wide resection of small intestine including the tumor, left hemicolectomy, left nephroureterectomy, splenectomy, partial pancreatectomy, duodeno-ileostomy and transverse sigmoidostomy was done on December 19, 1985. Pathological diagnosis was malignant lymphoma, diffuse small cell type infiltrating ureter, kidney and perirenal connective tissue. Because of poor postoperative course systemic chemotherapy was not performed and he died of disseminated intravascular coagulation on April 2, 1986. PMID- 3041778 TI - [Leiomyoma of the urinary bladder associated with right vesicoureteral reflux]. AB - A 32-year-old female consulted our hospital with the complaint of recurrent urinary tract infections, especially acute pyelonephritis. Cystoscopy revealed a wide-based tumor covered with a normal epithelium at internal meatus and right slight opened orifice, the contraction of which was slightly weak. Excretory urography showed almost normal nephrogram, pyelogram and ureterogram, and voiding cystography revealed right vesicoureteral reflux with grade IIb. Under general anesthesia, tumor resection of the bladder and right ureterovesiconeostomy were carried out. Pathologically the tumor was diagnosed leiomyoma. Right vesicoureteral reflux was speculated to have occurred secondarily to leiomyoma of the urinary bladder. PMID- 3041779 TI - [A case of pheochromocytoma of the urinary bladder]. AB - A case of pheochromocytoma of the urinary bladder is reported. A 17-year-old male was admitted to our hospital because of gross hematuria and urinary retention on Nov. 10, 1985. There was no history of hypertension. Intravenous pyelography and cystogram demonstrated a filling defect in the bladder. Transabdominal sonography disclosed a solid mass in the anterior wall and bladder neck. Cystoscopic examination revealed a non-papillary and broad based tumor in the anterior wall and bladder neck. Partial resection of the bladder wall was performed under the diagnosis of bladder tumor. The histological diagnosis was pheochromocytoma of the urinary bladder. PMID- 3041781 TI - [A case of urinary retention due to a female paraurethral leiomyoma]. AB - A case of urinary retention due to a female paraurethral leiomyoma is reported. The patient was a 67-year-old female who visited our clinic complaining of urinary retention. Urinary sediment and infravenous pyelography revealed no abnormal findings. An oval-shaped mass was palpated at the anterior vaginal wall. The tumor was 7 X 6 cm in size, elastic hard in consistency, irregular surfaced and well movable. Computed tomography revealed a calcified tumor in the urethrovaginal septum extending over all the urethral length from the bladder neck to the urethral meatus. Cystourethroscopic examination revealed a bladder neck elevation. Transvaginal needle biopsy was performed and the pathological diagnosis was benign leiomyoma. Transvaginal removal of the tumor was performed under the diagnosis of female paraurethral leiomyoma. The tumor was easily dissected from the vaginal wall but was adhered rather firmly to the urethra. The origin of the tumor could not be identified. Postoperatively, urination was improved. Female paraurethral leiomyoma is rare but must be considered as a cause of urinary retention. PMID- 3041780 TI - [A case of vesicosigmoidal fistula]. AB - The clinical course of a case of vesicosigmoidal fistula is presented. The patient, a 76-year-old woman, became aware of terminal micturition pain and pollakisuria in February, 1985. She was first treated under the diagnosis of cystitis to be relieved of her subjective symptoms, although there was no improvement of pyuria. She also began to feel lower abdominal pain on April 3, 1985. After various examinations including intravenous pyelography, enteroclysis and cystoscopy the diagnosis of vesicosigmoidal fistula originating from sigmoid diverticulitis was established. Careful observation at operation revealed remarkable adhesion among the sigmoid colon, bladder, uterus and left ovary. The sigmoid colon, was resected followed by end-to-end anastomosis. Because of considerably extensive inflammatory changes over the mucosal membrane of the bladder, the hole of fistula on the vesical wall was simply closed from outside of the bladder without performing partial cystectomy. Histological examination only demonstrated non-specific inflammatory changes without evidence of malignancy. She had a favorable progress postoperatively. PMID- 3041782 TI - [A case of adenomatous polyps with prostatic type epithelium]. AB - A 41-year-old male presented with gross hematuria and was found to have a polypoid lesion of the prostatic urethra. This proved to be an adenomatous polyp with prostatic type epithelium. Transurethral resection was performed on August 3, 1985. He had no evidence of recurrence following the operation. Previously reported cases of relevance are briefly discussed. PMID- 3041783 TI - [Lipogranuloma with marked eosinophile infiltration in male genitalia]. AB - A characteristic lipogranuloma in the male genitalia is presented. Histopathological examination revealed a granuloma resembling sclerosing lipogranuloma with marked eosinophile infiltration. Only 11 cases with such a histopathological feature have been reported. Including this case all cases have similar localization, shape and clinical course. The genesis of these granulomas is discussed. PMID- 3041784 TI - [A case of intrascrotal leiomyoma originating from tunica vaginalis]. AB - A case of intrascrotal leiomyoma originating from tunica vaginalis is reported. The patient was a 63-year-old male who complained of painless tumor of the right scrotal contents. The tumor was surgically removed easily. The histological examination of the tumor revealed leiomyoma. Seven cases of intrascrotal leiomyoma reported in Japan were reviewed. PMID- 3041786 TI - The annual oration for 1888. Medical Association of the State of Alabama. By Benjamin James Baldwin. PMID- 3041785 TI - [Clinical effects of terodiline hydrochloride on urinary frequency and sense of residual urine--a double blind clinical trial using flavoxate hydrochloride as a control]. AB - A double blind clinical trial was performed as a multicenter study to determine the usefulness of terodiline hydrochloride (HCl), an anticholinergic and calcium antagonistic agent, for urinary frequency or sense of residual urine in patients with psychogenic diseases, chronic prostatitis or chronic cystitis. Either 24 mg of terodiline HCl a day or 600 mg of flavoxate HCl a day was given for 4 weeks. One hundred and ninety-nine patients completed the test. The final global improvement rating was 70% in patients given terodiline HCl and 48% in patients given flavoxate HCl. The difference was statistically significant (p less than 0.01). Diurnal and nocturnal urinary frequency and urinary incontinence were less in patients given terodiline HCl than in patients given flavoxate HCl (p less than 0.01). No difference was noted between the two agents in relieving sense of residual urine. Compared with the control period, the average urinary frequency decreased 2.0 times a day in patients given terodiline HCl and 0.7 times in patients given flavoxate HCl. The difference was statistically significant (p less than 0.01). Adverse effects were observed in 12% of the patients given terodiline HCl and in 16% of the patients given flavoxate HCl. They included thirst, difficult urination, constipation, slight increase of serum GOT, GPT or alkaline phosphatase, and so forth. They disappeared with discontinued use of the agent. The global utility rating was 68% in patients given terodiline HCl and 45% in patients given flavoxate HCl, the difference being significant (p less than 0.01). These results indicate that terodiline HCl is useful for the treatment of urinary symptoms in patients with psychogenic diseases, chronic prostatitis or chronic cystitis. PMID- 3041787 TI - [A monoclonal antibody against peripheral human blood monocytes (RoMo-1)]. AB - The monoclonal IgG2a-antibody RoMo-1 binds to peripheral human blood monocytes but does not react with other blood cells, tissue macrophages, HL-60, K 562 or U 937. It seems possible, that the antibody defines an antigen expressed on the premonocytic and monocyte stages of cells from the mononuclear phagocytic system only. The antibody is cytotoxic. PMID- 3041788 TI - Effects of long-term combined dosing with nicardipine and propranolol on coronary hemodynamics, myocardial metabolism, and exercise tolerance in patients with angina pectoris: comparison with monotherapy. AB - To determine whether the association of nicardipine with propranolol had additive effects on myocardial metabolism, 16 patients with angina pectoris were studied invasively before and after 1 month of therapy with a combination of nicardipine and propranolol and compared to a group of 42 patients treated with nicardipine (n = 17) or propranolol (n = 25) alone. When data were compared at a fixed heart rate (atrial pacing), mean blood pressure was reduced with combined treatment from 96 +/- 19 to 76 +/- 13 mm Hg (p less than 0.003). Myocardial oxygen uptake and coronary sinus flow decreased significantly from 20 +/- 9 to 14 +/- 6 ml/min (p less than 0.015) and from 152 to 111 ml/min (p less than 0.05), respectively. The arterio-coronary sinus difference in oxygen content also decreased (13.3 to 12.5 ml/dl; p less than 0.0025), suggesting an improved balance between oxygen supply and demand. Such changes in coronary blood flow and myocardial oxygen uptake were not observed in the group of patients assigned to monotherapy. Lactate uptake rose and the abnormal glutamine production, which worsened with propranolol monotherapy, improved with nicardipine and propranolol (-2.0 to -1.4 mumol/min; p less than 0.05 vs propranolol). The superiority of nicardipine and propranolol over propranolol monotherapy was maintained during a pacing stress test. Thus the combination of nicardipine with a beta blocker had greater oxygen sparing effects and restored aerobic metabolism better than either drug alone, allowing optimal use of the coronary reserve. PMID- 3041789 TI - Effects of acute intrathoracic pressure changes on left ventricular geometry and filling. AB - Acute changes in intrathoracic pressure (ITP) affect left ventricular (LV) function. It has been suggested that this functional impairment could be the result of an alteration in LV filling caused by a reduction in LV compliance induced by the rearrangement of biventricular geometry that occurs under these conditions. Therefore, to evaluate the effects of an acute increase or decrease in ITP on LV geometry and filling, we used two-dimensional and Doppler echocardiography to study 25 normal volunteers both during the Muller maneuver (acute decrease in ITP induced by a forced inspiration against a closed airway) and during continuous positive airway pressure breathing. During both maneuvers LV geometry was altered as demonstrated by the significant increase in the normalized curvature radius of the interventricular septum and the unchanged curvature radius of the LV free wall. LV filling was altered during both maneuvers as demonstrated by significant decreases in early peak flow velocity, early-to-late peak flow velocity ratio, and early deceleration rate. Thus, during maneuvers that acutely decrease or increase ITP, alterations in LV geometry occur. These acute distortions of LV geometry may be one of the mechanisms responsible for alterations in LV filling. PMID- 3041790 TI - The diagnostic and prognostic value of the treadmill exercise test in the evaluation of chest pain, in patients with recent myocardial infarction, and in asymptomatic individuals. PMID- 3041791 TI - An impression of Alice Hamilton--physician, industrial hygienist, crusader and friend. PMID- 3041792 TI - Gross and histologic anatomy of total anomalous pulmonary venous connections. AB - Among 49 heart specimens with total anomalous pulmonary venous connection (TAPVC), obstruction to pulmonary venous flow was present in all 13 cases with TAPVC below the diaphragm and in 53% of 36 cases with TAPVC above the diaphragm. Obstruction was produced by extrinsic pressures on the vein, intrinsic narrowing of the vein, or both. The histology of the narrowed veins was extremely variable, ranging from atrophy of the vein wall to hypertrophy of intima, media-adventitia, or both. Balloon dilation of narrowed veins was performed in 3 cases, without clinical or anatomic evidence of success. PMID- 3041793 TI - Verapamil versus acebutolol for syndrome X. PMID- 3041795 TI - Sutures and forces: a review. AB - This review gives a description of the biologic significance of craniofacial sutures with respect to growth and to growth corrections. Sutural growth and its regulation are discussed briefly. Morphogenesis of sutures, sutural morphology, both microscopic and macroscopic, the structure and function of the sutural periosteum and secondary cartilages, and the biochemical composition of sutures are described. Furthermore, in vivo and in vitro experiments, including transplantation experiments, are discussed. The relationship between extrinsic mechanical forces and the resulting tissue responses in sutures is given special attention. The present article describes the state of our knowledge on the interaction between sutures and forces, and indicates problems that need to be investigated. PMID- 3041794 TI - Successful management of acute myocarditis with biventricular assist devices and cardiac transplantation. PMID- 3041796 TI - Clinical management of the acrylic splint Herbst appliance. AB - This article describes one variation in Herbst appliance design--the acrylic splint Herbst. Topics discussed include early fixed appliance treatment before Herbst therapy, impression taking, bite registration, and evaluation of appliance fabrication. The specifics of appliance delivery, including bonding of the maxillary appliance when indicated, are also discussed as are the techniques of appliance advancement and removal. PMID- 3041797 TI - Techniques for improving orthodontic results in the treatment of missing maxillary lateral incisors. A case report with literature review. AB - We have presented a case report of the orthodontic treatment of a patient with congenitally missing lateral incisors and with skeletal and dental deep bite. Information was provided on methods to improve clinical orthodontic results. Different sections deal with (1) various diagnostic criteria for the best treatment approach, (2) variations in maxillary arch wire manipulation, (3) biomechanical considerations, (4) modifications of clinical crown by special procedures so that the canines resemble and function as lateral incisors, (5) functional considerations of the occlusion, and (6) details in finishing. Particular emphasis was placed on the use of a special design of closing loops for simultaneous space closure and intrusion of the anterior teeth. This article has discussed the principles of incisor and canine intrusion, and has demonstrated that the closing loops described are capable of intruding incisors with minimal side effects on the posterior teeth. It has also demonstrated how control of the mechanical variables dramatically increased the efficiency and effectiveness of intra- and extraoral forces in the treatment of malocclusion. PMID- 3041798 TI - Not all appliances are recreated equal... PMID- 3041799 TI - Linear C3 deposits on the tubular basement membrane in renal allograft biopsies. AB - Linear and granular tubular basement membrane (TBM) deposits of C3 occur in renal allograft biopsies, but their significance is unknown. We retrospectively analyzed the predictive importance of C3 deposits in 88 biopsied transplant patients with allograft dysfunction. All patients were followed for greater than or equal to 2 years from biopsy. Patients were divided into three groups: group I: no C3 deposits, 47 patients; group II: granular deposits of C3, 28 patients; and group III: linear TBM deposits, 13 patients. The incidence of acute and chronic rejection was not different. In group III, 12 grafts were lost by 5 years (92%), and the remaining patient has chronic rejection. Group III survival was significantly less than groups I and II (Kaplan-Meier curves), P = 0.02, but graft survival in groups I and II were similar. There was no association of anti TBM antibody deposits with C3, and the mechanism of deposition is unknown. We conclude that the presence of linear C3 deposits along the TBM in the setting of allograft dysfunction is associated with decreased allograft survival. PMID- 3041800 TI - The clinical spectrum of renal disease associated with human immunodeficiency virus. AB - A nephrology consultation was called on 100 adult patients of 1,635 (6.1%) patients with human immunodeficiency virus (HIV) infection seen between 1982 and 1987 at the University of Miami/Jackson Memorial Medical Center. Renal disease was observed in all groups of patients with a risk factor for HIV infection with a lesser incidence, however, among homosexuals. Intravenous drug (IVD) use and possibly race appear to be important factors in the development of renal complications. Renal disease was the dominant clinical feature in eight asymptomatic HIV carriers and in 34 patients with AIDS-related complex (ARC) who had not developed the opportunistic infections and/or malignancies associated with acquired immunodeficiency syndrome (AIDS). Ninety-one percent of consultations were requested for evaluation of proteinuria and/or renal failure. Nephrotic range proteinuria, in excess of 3 g/24 h, was present in 52 patients, and was less prevalent in homosexuals than in other groups at risk. Renal failure (serum creatinine greater than or equal to 5 mg/dL), initially present in 32 patients, eventually developed in 69 and improved in only 18 of them. A renal biopsy, obtained for work-up of nephrotic syndrome (22 patients) or renal insufficiency (3 patients), uncovered a picture of focal and segmental glomerulosclerosis in all 25 instances. Overall, 76 patients are dead, seven are lost to follow-up, and 17 are alive, of whom eight (four HIV carriers, two patients with ARC, and two with AIDS) are on maintenance hemodialysis with a mean survival time of 217 days. PMID- 3041801 TI - Recurrent IgA nephropathy in living-related donor transplantation: recurrence or transmission of familial disease? AB - We describe a patient who developed terminal renal failure of two HLA-identical renal allografts due to crescentic IgA nephropathy. The first graft contained IgA deposits at the time of donation, suggesting that transmission of IgA deposits may have contributed to the nephritis of the first allograft. The second graft was free of IgA deposits at the time of donation, but the recipient developed a similar, rapidly progressive nephritis. This case points up the malignant potential of IgA nephropathy and the complex nature of transplant planning for patients with end-stage renal disease (ESRD) secondary to IgA nephropathy. Living related donor (LRD) transplants seem to be associated with a higher rate of recurrence than cadaveric grafts. This higher rate may partly reflect the inadvertent transmission of subclinical IgA deposits from donor to recipient and a genetic susceptibility of certain HLA types (specifically B35 and DR4) to recurrent disease. Cadaveric transplants may be preferable in the setting of high risk HLA types or familial patterns of IgA nephropathy. PMID- 3041802 TI - Platelet activation and prostacyclin supporting capacity in the loin pain hematuria syndrome. AB - The loin pain hematuria syndrome has been characterized as a constellation of severe recurrent flank pain and hematuria, occurring predominantly in young women. We studied a 17-year-old woman who had recurrent right flank pain, gross hematuria, and fever, without evidence of urinary tract infection. Her physical exam was remarkable for right costovertebral angle tenderness and a normal BP. Her urinalysis showed blood and protein but her creatinine clearance and 24-hour urinary calcium excretion were normal. A kidney biopsy was remarkable for arteriolar subintimal fibrous thickening and fibrin deposition, but no glomerulonephritis. Her peripheral hemostasis evaluation was normal except for circulating platelet aggregates and elevated fibrinopeptide A levels. On two occasions, her serum was unable to normally support prostacyclin (PGI2) production by cultured human umbilical endothelial cells, as measured by radioimmunoassay (RIA) of its stable metabolite 6-keto-PGF alpha. Blood samples from the right renal vein and inferior vena cava revealed a selective elevation of fibrinopeptide A in the right renal venous effluent. The presence of circulating platelet aggregates and elevated levels of fibrinopeptide A (a cleavage product of fibrin) suggests that platelet activation and fibrin deposition may play a role in the pathogenesis of this disorder. The inability of her serum to normally support the production of the potent antiplatelet and antithrombotic substance, PGI2, could represent a primary renovascular endothelial cell defect. PMID- 3041803 TI - Recurrence of disease following renal transplantation. AB - The diagnosis of recurrent renal disease after transplantation is dependent on an accurate and complete diagnosis of the initial cause of renal failure and a similar determination of the cause of graft failure. To be classified as recurrent, the disease in the renal graft must be identical to that seen in the native kidneys. Recurrence of disease accounts for less than 2% of all graft failures, but the overall incidence of recurrent disease is probably 5 to 10 times more common. The most frequent cause of recurrent disease is glomerulonephritis, which was first recognized to recur soon after renal transplantation was introduced. It was then recognized that a variety of metabolic disorders would recur, but it has taken 25 years of experience for a clear picture to emerge of recurrence in most conditions. No initial cause of renal failure poses a contraindication to at least one attempt at transplantation, although with Fabry's disease and oxalosis, a special assessment of the risks for the individual recipient is warranted. In some patients, experience has shown the need for a delay in the commitment to transplantation (eg, in those with anti-glomerular basement membrane [GBM] antibody glomerulonephritis or Henoch Schonlein purpura), the need for the choice of a particular immunosuppressive regimen (eg, in hemolytic uremic syndrome [HUS]), the need for avoidance of primary nonfunction (eg, in oxalosis), and the desirability of avoiding live kidney donation (eg, in heterozygote donors in Fabry's disease, high-risk recipients with focal glomerulosclerosis, and in recipients with HUS). Probably all types of glomerulonephritis recur, but with great variation in frequency and severity. In some forms of glomerulonephritis, recurrence may be frequent and definite on histopathological criteria but may only have a minor clinical expression (eg, dense deposit disease, anti-GBM antibody glomerulonephritis, IgA nephropathy), but in others, recurrence is less predictable yet it is clearly associated with premature graft failure (eg, focal glomerulosclerosis, membranous nephropathy). A common theme emerging is that where the initial glomerulonephritis is aggressive and causes kidney failure over a short time, recurrence is more likely, and when present, it will lead to graft failure with an increased frequency. Clinical manifestations, the frequency of recurrence, and the prognosis of the graft are now identified for most conditions. Unexpected observations have included the rarity of recurrent systemic lupus erythematosus (SLE), the immediate return of heavy proteinuria in focal glomerulosclerosis, and the predictable return of dense deposit disease.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3041805 TI - Unresolved issues in the future of pharmacy. AB - The effect of the major forces shaping pharmacy's future--technology, economics, and social values--is discussed. Pharmacy can best respond to these forces by expanding the informational component of pharmacy practice and by returning to its fundamental relationship with society--that is, by accepting responsibility for drug-use control. To accomplish this, pharmacy must go through a process of occupational reconstruction and self-renewal. Technological forces shaping pharmacy practice include computers and robotics, communications, and therapeutics. Regarding economic changes, medical-care decisions are shifting away from individuals toward "third parties." This factor, combined with output based payment systems, may lead to a consolidation of service providers. Marketing pharmaceutical services and organizing pharmacy's internal structure will become important. Some of the social forces affecting pharmacy's future include the aging population, the weakening of professional authority, and pharmacy's public image. Just as society goes through the "information revolution," pharmacy will need to undergo a "reprofessionalization." Pharmacy's societal function should involve "pharmaceutical care," a concept that includes both drug-product control and clinical pharmacy services. Pharmacy should return to its preindustrial origins in valued, complex, specific, and committed public service if it wishes a happy postindustrial future. PMID- 3041804 TI - The 6's and 17's of developmental mutants near the major histocompatibility complex: the mouse t-complex does not have a human equivalent. PMID- 3041806 TI - Pharmacy policies in the Arizona prepaid Medicaid program. PMID- 3041807 TI - Home testing products for self-monitoring. AB - Currently available newer home testing products and some that may be available in the near future are described. Although not diagnostic, home testing products may be used by consumers to recognize medical conditions early so that a physician may be consulted. Common conditions or situations that can be monitored or tested include blood glucose concentrations, pregnancy, ovulation, urinary-tract infection, the presence of fecal occult blood, and asthma. To minimize the possibility of false-positive and false-negative results and to ensure that consumers understand how to use the products correctly, pharmacists should discuss the tests and their instructions with users beforehand. When home testing products are used according to instructions, most provide accurate results (95% 99%). Home testing products will continue to be developed, using monoclonal antibodies as their base of detection. Pharmacists can play an important role in ensuring that consumers use these products correctly. PMID- 3041808 TI - Factors affecting sensitivity and specificity of a diagnostic test: the exercise thallium scintigram. AB - Technical and methodological factors might affect the reported accuracies of diagnostic tests. To assess their influence on the accuracy of exercise thallium scintigraphy, the medical literature (1977 to 1986) was non-selectively searched and meta-analysis was applied to the 56 publications thus retrieved. These were analyzed for year of publication, sex and mean age of patients, percentage of patients with angina pectoris, percentage of patients with prior myocardial infarction, percentage of patients taking beta-blocking medications, and for angiographic referral (workup) bias, blinding of tests, and technical factors. The percentage of patients with myocardial infarction had the highest correlation with sensitivity (0.45, p = 0.0007). Only the inclusion of subjects with prior infarction and the percentage of men in the study group were independently and significantly (p less than 0.05) related to test sensitivity. Both the presence of workup bias and publication year adversely affected specificity (p less than 0.05). Of these two factors, publication year had the strongest association by stepwise linear regression. This analysis suggests that the reported sensitivity of thallium scintigraphy is higher and the specificity lower than that expected in clinical practice because of the presence of workup bias and the inappropriate inclusion of post-infarct patients. PMID- 3041809 TI - Single kidney function: effect of acute protein and water loading on microalbuminuria. AB - The hyperfiltration induced by an acute response to an oral protein and water load was investigated to ascertain whether it can modify the urinary albumin excretion (UAE) in the microalbuminuric range by further increasing the glomerular filter permeability. To this end, six patients with a single kidney selected as having microalbuminuria on a regular diet without the clinical or laboratory data of overt renal disease and eight healthy subjects received a short-term protein and water load (150 g of meat-derived protein and 1 liter of water). In patients with one kidney, mean basal UAE values were significantly higher than in control subjects (p less than 0.006), whereas endogenous creatinine clearance values were only slightly lower (p greater than 0.05). One hour after the protein and water load, an abrupt increase in microalbuminuria levels was found in patients with one kidney and mean UAE values were significantly higher than in control subjects (p less than 0.002), whereas mean creatinine clearance values were significantly lower in patients than in control subjects (p less than 0.01). High UAE (p less than 0.002) and low creatinine clearance (p less than 0.002) values were maintained over the following four hours in patients with one kidney. These data suggest that in the single kidney with reduced renal functional reserve, an oral protein and water load magnifies the pre-existing loss of glomerular permselective properties due to chronic hyperfiltration as manifested by a further increase in microalbuminuria. PMID- 3041810 TI - Goodpasture's syndrome: recurrence after a five-year remission. Case report and review of the literature. AB - Herein is reported the case of a man who has had a recurrence of Goodpasture's syndrome following a five-year remission. The patient presented initially in 1977 at the age of 28 with Goodpasture's syndrome manifested by pulmonary hemorrhage without clinical evidence of renal disease, and positive antiglomerular basement membrane antibody. Following treatment with corticosteroids, remission occurred and the serum antiglomerular basement membrane antibody became negative. In 1983, he experienced a relapse with the reappearance of serum antiglomerular basement membrane antibody, the development of severe life-threatening intrapulmonary hemorrhage, and hematuria. This case illustrates that life-threatening relapse may occur in Goodpasture's syndrome despite a prolonged remission and the disappearance of detectable antiglomerular basement membrane antibody in the circulation. PMID- 3041811 TI - Acute paraplegia: a presenting manifestation of aortic dissection. AB - Two patients who presented with acute paralysis of the lower extremities as an initial manifestation of aortic dissection are described. The first patient had transient chest pain followed by flaccid paralysis of her lower extremities and severe back pain. In the second patient, sudden paralysis of both legs developed without pain of any sort. The paraplegia completely resolved in a few minutes; however, chest and back pain later ensued. Both patients had a proximal (type I or A) aortic dissection. The first patient's entrance tear in the aortic intima was just above the aortic valve with antegrade propagation, whereas in the second patient, the entrance tear was at the aortic isthmus, with both antegrade and retrograde dissection. Acute cardiac tamponade resulted in sudden deterioration and death in both patients, before any therapeutic intervention could be entertained. PMID- 3041812 TI - Histochemical demonstration of iron but not aluminum in a case of dialysis associated osteomalacia. AB - A patient undergoing hemodialysis is described in whom osteomalacia developed despite protracted treatment with calcitriol. Appropriately stained biopsy sections exhibited iron at all marrow-osteoid interfaces and a small fraction of trabecular mineralization fronts. Aluminum, the metal usually associated with osteomalacia in patients undergoing hemodialysis, was not histochemically demonstrable, even though spectrophotometrically measured bone aluminum content was substantial. These observations suggest two interpretations: iron may have caused osteomalacia through effects on bone cells and at mineralization fronts; alternatively, aluminum may have caused osteomalacia while remaining histochemically undetectable. It is possible that both metals exerted toxic effects simultaneously. PMID- 3041813 TI - Factitious dermatosis masquerading as recurrent herpes zoster. AB - A 35-year-old nurse's aide presented with monthly episodes, during her menses, of self-induced cutaneous lesions intended to simulate recurrent herpes zoster. Features of the clinical presentation that prompted the correct diagnosis are discussed. PMID- 3041814 TI - Poor survival of patients with idiopathic cardiomyopathy considered too well for transplantation. PMID- 3041815 TI - Controlled trials of double beta-lactam therapy with cefoperazone plus piperacillin in febrile granulocytopenic patients. AB - The efficacy and safety of double beta-lactam therapy with cefoperazone plus piperacillin in febrile granulocytopenic patients were compared with moxalactam plus piperacillin, ceftazidime plus piperacillin, and imipenem alone in two separate clinical trials. All patients also received prophylactic vitamin K. When National Committee for Clinical Laboratory Standards breakpoints for susceptibility were used, a greater proportion of pretherapy isolates of gram negative aerobic bacilli and gram-positive organisms were found to be susceptible to cefoperazone (94 percent) and imipenem (91 percent) than to moxalactam (84 percent), ceftazidime (85 percent), or piperacillin (85 percent). In trial I, the overall response rates for documented or possible infections were 78 percent (76 of 97 patients) for cefoperazone/piperacillin and 80 percent (72 of 90 patients) for moxalactam/piperacillin. In trial II, the overall response rates were 86 percent (25 of 29 patients) for cefoperazone/piperacillin, 74 percent (20 of 27 patients) for ceftazidime/piperacillin, and 72 percent (21 of 29 patients) for imipenem alone. There was no nephrotoxicity or hemorrhage related to the study drugs. Diarrhea was more frequent with each of the double beta-lactam regimens, whereas nausea and seizures were more common with imipenem given at a dosage of 1.0 g intravenously every six hours. Seizures occurred in three of 29 imipenem treated patients but in none of 243 patients treated with the double beta-lactam regimens (p less than 0.001). These results suggest that cefoperazone plus piperacillin provides adequate coverage for most common bacterial pathogens and is safe and effective therapy for febrile granulocytopenic patients. PMID- 3041816 TI - Cefoperazone plus tobramycin versus ticarcillin plus tobramycin in febrile granulocytopenic cancer patients. AB - Cefoperazone plus tobramycin was compared in a prospective, randomized trial with our standard regimen of ticarcillin plus tobramycin as empiric therapy of fever in granulocytopenic patients with cancer. Patients who received cefoperazone were also given vitamin K (5 mg orally twice a week). Of 39 microbiologically and clinically documented infections treated with ticarcillin plus tobramycin, 28 (72 percent) showed improvement. Of 27 microbiologically and clinically documented infections treated with cefoperazone plus tobramycin, 21 (78 percent) showed improvement. The overall response rates were similar (40 of 53, or 74 percent, for ticarcillin plus tobramycin versus 38 of 48, or 79 percent for cefoperazone plus tobramycin). There was no difference in response between groups according to site of infection. Serious side effects were minimal with both regimens. There were no enterococcal superinfections in patients receiving cefoperazone. These results suggest that the overall efficacy and toxicity of study regimens are similar. PMID- 3041817 TI - Cefoperazone plus piperacillin versus mezlocillin plus tobramycin as empiric therapy for febrile episodes in neutropenic patients. AB - The double beta-lactam combination of cefoperazone plus piperacillin was compared with an aminoglycoside-containing regimen of mezlocillin plus tobramycin in a prospective, randomized trial of empiric therapy for febrile neutropenic patients (neutrophils no more than 1,000/mm3). Thirty febrile episodes were treated with cefoperazone plus piperacillin and mezlocillin plus tobramycin, respectively. There was no significant difference between the two groups with respect to age, sex, pretherapy neutrophil count, and mean duration of therapy. The majority of patients had neutrophil counts of no more than 200/mm3 at the initiation of therapy. Only microbiologically and clinically documented infections were evaluated for efficacy. The cefoperazone plus piperacillin regimen appeared to have a comparable response rate with the mezlocillin plus tobramycin regimen (20 of 24 patients [83 percent] versus 16 of 23 patients [70 percent]). Gram-positive micro-organisms were seen predominantly in this study, with the cefoperazone plus piperacillin regimen achieving a bacteriologic response in 84 percent, as opposed to 60 percent for those organisms treated with the mezlocillin plus tobramycin regimen. Neither regimen was totally effective against coagulase-negative staphylococci. Eight superinfections occurred in the cefoperazone plus piperacillin arm, whereas 11 superinfections occurred in the mezlocillin plus tobramycin arm. Although fungal superinfections were most common, the number of gram-positive superinfections in the mezlocillin plus tobramycin arm exceeded those seen in the cefoperazone plus piperacillin arm. The incidence of antibiotic related side effects was similar in the two groups. Hypokalemia was most frequently seen. Both skin rashes and nephrotoxicity were more common with mezlocillin plus tobramycin. Cefoperazone plus piperacillin was found to be effective empiric therapy in febrile neutropenic patients. This double beta lactam combination may be particularly useful for patients who have or are at high risk for the development of renal insufficiency. PMID- 3041819 TI - Survey of antibiotic susceptibility among gram-negative bacilli at a cancer hospital. AB - A survey was conducted of the susceptibility of gram-negative bacilli to selected broad-spectrum antibiotics. The organisms were isolated from all patient specimens submitted to the routine microbiology laboratory during two three-month periods. Overall, the least resistance was observed against imipenem and ciprofloxacin. Considering all of the gram-negative bacilli, differences in susceptibilities to the other antibiotics (aztreonam, cefoperazone, ceftazidime, piperacillin) were minimal. Significant increases in resistance to some antibiotics occurred during the latter period. PMID- 3041818 TI - Cefoperazone versus ceftazidime monotherapy of nosocomial pneumonia. AB - Cefoperazone and ceftazidime monotherapy were compared in a randomized, prospective evaluation of patients with nosocomial pneumonia. These antibiotics were equally effective, with an overall successful treatment rate of 45 of 62 (73 percent) for cefoperazone-treated patients and 50 of 63 (79 percent) for ceftazidime-treated patients (p = 0.41). There was no difference in the incidence of side effects (including hypoprothrombinemia), superinfections, or colonization of the oropharynx with yeast, enterococcus, Staphylococcus aureus, or resistant gram-negative bacilli. When antibiotic administration, and laboratory costs are considered, cefoperazone is less expensive than ceftazidime. Both cefoperazone and ceftazidime are effective therapy for nosocomial pneumonia. PMID- 3041820 TI - Cefoperazone in the treatment of infections in cancer patients. AB - Cefoperazone appears to be one of the cephalosporins with the most promise in granulocytopenic patients, regardless of whether it is used in combination or as monotherapy. It has the broad spectrum of antimicrobial activity necessary to inhibit the organisms commonly encountered in patients with cancer, as well as the ability to achieve high serum levels for sustained periods of time. Administration of cefoperazone should provide effective therapy in the overall management of most infections encountered in patients with neoplastic diseases. PMID- 3041821 TI - Multicenter comparison of once-daily Uniphyl tablets administered in the morning or evening with baseline twice-daily theophylline therapy in patients with nocturnal asthma. AB - Ninety-six patients with reversible airways disease and a history of nocturnal asthma completed a four-week open-label study that compared the effectiveness of Uniphyl tablets (The Purdue Frederick Company, Norwalk, Connecticut) administered once daily in the morning or the evening with twice-daily baseline theophylline therapy. All patients transferred easily from their previous twice-daily theophylline regimen to once-daily Uniphyl therapy. Predose serum theophylline levels and pulmonary function values were similar for the twice-daily and once daily theophylline regimens. However, both the morning and evening once-daily Uniphyl regimens were judged by the investigators to control the asthmatic symptoms of more patients than did the prestudy twice-daily theophylline. In addition, patient acceptance of the two once-daily Uniphyl regimens was significantly (p less than 0.01) greater than that of the previous twice-daily theophylline therapy. During the fourth week of the study, the evening Uniphyl dosing schedule resulted in significantly (p less than 0.01) higher home monitored morning peak expiratory flow rates and fewer nighttime awakenings than the pre-study twice-daily theophylline regimen. The morning regimen was not associated with such an improvement. The results of the study suggest that a once daily evening dosage regimen with Uniphyl tablets provides greater control of nocturnal asthma. PMID- 3041822 TI - Is a uniform round-the-clock theophylline blood level necessary for optimal asthma therapy in the adolescent patient? AB - Twenty-one patients, 12 to 18 years of age, with nocturnal asthma controlled with sustained-release theophylline administered twice daily, were enrolled in a 10 week, double-blind, two-way crossover study that compared Theo-Dur tablets administered twice daily with an equivalent daily dose of Uniphyl tablets administered once daily at bedtime. Seventeen patients completed the study. The mean morning theophylline serum level obtained with Uniphyl tablets was significantly higher than that obtained with Theo-Dur tablets (13.1 versus 9.6 micrograms/ml, p = 0.02). The mean evening serum level was significantly lower with Uniphyl tablets (6.3 versus 10.1 micrograms/ml, p = 0.003). Despite these differences in serum concentrations, morning and evening pulmonary function test values (forced expiratory volume in one second and peak expiratory flow rate) and symptom scores were nearly identical for the two preparations, as was the supplemental use of aerosol bronchodilators. Once-daily dosing with Uniphyl tablets may benefit adolescent patients with nocturnal asthma by increasing compliance and providing better asthma control. In addition, the lower daytime theophylline levels produced by this preparation may also reduce long-term adverse effects on behavior and cognition. PMID- 3041823 TI - Circadian rhythms. AB - All forms of life, from the simplest cells to the most complex organisms, show periodicity in some of their biologic activities and functions. The most common of these rhythmic events are those that we refer to as "circadian" (circa, around; dias, day). In humans, the caliber of both the upper and lower airways shows circadian fluctuation that is amplified in disease states. The caliber of the airways of the tracheobronchial tree decreases at night and increases during the day. In asthmatic persons, the nocturnal decrease is amplified, causing peak dyspnea, wheezing, cough, sneezing, rhinorrhea, and nasal stuffiness to occur between 2:00 and 6:00 A.M. The most effective pharmacologic strategies for the treatment of these symptoms appear to be those timed to provide maximal medication between these hours, when it is needed the most. PMID- 3041824 TI - Improved control of asthma in the office setting. A large-scale study of once daily evening doses of theophylline. AB - A large-scale, multi-investigator open evaluation compared a once-daily regimen of controlled-release theophylline (Uniphyl tablets) with previous twice- or thrice-daily methylxanthine regimens. Three hundred asthmatic patients, 78 percent prone to nocturnal episodes during prior therapy, completed the investigation. Eighty-two percent of the patients were treated for moderate or severe disease. After a one-week evaluation of baseline theophylline therapy (with adjunctive medication), the patients substituted evening doses of the once daily drug in approximate milligram-for-milligram equivalent doses. Concomitant medications were allowed as before. Nighttime and morning asthma control improved significantly without deterioration in the evening, and without increased side effects. Once-daily therapy resulted in markedly fewer night awakenings involving inhaler use (p less than 0.01), and near 60 percent reductions in the number of patients with nighttime or early morning exacerbations (p less than 0.01). Control of morning chest tightness, wheeze, and dyspnea improved significantly (p less than 0.01), and patients' as well as investigators' global evaluations favored once-daily treatment (p less than 0.01). Morning peak expiratory flow rates improved both at home (p less than 0.01) and at the office (p = 0.05). The forced expiratory volume in one second at the office increased modestly in the entire group. It is concluded that Uniphyl is effective and well tolerated when administered in once-daily evening doses. PMID- 3041825 TI - Biologic rhythms and medicine. AB - Medical practice and research in the United States and other countries are pervaded by the traditional concepts of homeostasis. As a result, the time of day or year when a diagnostic or therapeutic procedure is performed or a drug is administered is not considered important. However, during the last three decades, the science of chronobiology has revealed the importance to medicine of the rhythms that exist in the biologic functions and processes of humans. This article reviews the basic concepts of human chronobiology and discusses the rhythm dependence of certain disease states and the pharmacodynamics of medications frequently prescribed for their treatment. Special reference is made to the nocturnal exacerbation of asthma and the use of chronobiologic principles in its treatment. PMID- 3041826 TI - Safety and efficacy of once-daily Uniphyl tablets compared with twice-daily Theo Dur tablets in elderly patients with chronic airflow obstruction. AB - Oxygen desaturation and subclinical coronary artery disease may predispose older patients with chronic airflow obstruction to cardiac arrhythmias, especially when high concentrations of theophylline are present in the blood. To assess the safety and efficacy of Uniphyl tablets, an oral theophylline preparation formulated for once-daily dosing, in elderly patients with chronic airflow obstruction, we conducted a randomized, three-phase, double-blind crossover study comparing evening dosing with Uniphyl tablets, Theo-Dur tablets administered twice daily, and placebo. The patients in the study were scheduled to receive each treatment for two weeks. Each day, symptoms, side effects, peak expiratory flow rates, and use of metered-dose inhalers were recorded. Near the end of each phase, serum theophylline concentrations were measured every two hours between 8:00 A.M. and 8:00 P.M. on two consecutive days. The patients underwent ambulatory Holter monitoring during the final 48 hours of each phase. Twelve patients completed the active-drug phases of the study, but seven of the 12 were removed from the placebo phase because of increasing symptom severity. The difference between the number of patients completing the active-drug and placebo phases was statistically significant (p less than 0.001). Treatment with Uniphyl tablets resulted in a significantly (p less than 0.05) greater increase in peak expiratory flow rate than Theo-Dur tablet therapy, and both active drugs increased peak expiratory flow rate more than placebo. Circadian variation in peak expiratory flow rate was seen during the placebo and Theo-Dur tablet phases but not during the Uniphyl tablet phase. Symptoms and side effects were similar during the two active-drug phases. Cardiac ectopy was observed in most of the patients, but it was not significantly greater during the theophylline phases than during the placebo phase. Furthermore, ectopic activity was not directly related to the times of maximal serum theophylline concentration. PMID- 3041827 TI - Inflammatory mechanisms and nocturnal asthma. AB - The results of recent research strongly suggest that airway inflammation, which may increase at night as a result of circadian troughs in blood epinephrine and cortisol concentrations, underlies the bronchial hyperresponsiveness that is almost certainly a major contributor to the pathogenesis of nocturnal asthma. This article reviews what is known about the nature and complex interactions of the inflammatory cells and mediators that may be involved in asthma, with particular emphasis on nocturnal asthma. The roles of platelet-activating factor antagonists, corticosteroids, and theophylline in suppressing this response also are discussed. PMID- 3041828 TI - Hypertension in cyclosporine-treated renal transplant recipients is sodium dependent. AB - PURPOSE: Physicians increasingly prescribe cyclosporine as an immunosuppressive agent for both organ-transplant and non-organ-transplant recipients. Investigators have reported a high incidence of drug-induced hypertension even when clinical nephrotoxicity was not present. We wanted to determine the reason. PATIENTS AND METHODS: A comparison was made of hypertension in 15 cyclosporine treated transplant recipients with that in a similar group of 15 azathioprine treated transplant recipients. RESULTS: Hypertension in the cyclosporine group responded differently from that seen in the azathioprine group and from previously described forms of post-transplantation hypertension. Hypertensive cyclosporine-treated patients show a sodium acquisitive renal state that responds to sodium restriction. Unlike rat models, which suggest cyclosporine-induced stimulation of the renin-angiotensin system, or previous forms of post-transplant hypertension in humans, plasma renin levels were not elevated and blood pressure did not respond to a test dose of captopril. CONCLUSION: Hypertension in cyclosporine-treated patients is an iatrogenic form of hypertension that may be associated with an early, subtle, renal defect in sodium excretion, a genesis of hypertension that is consistent with Guyton's view of essential hypertension. PMID- 3041829 TI - Recombinant interferon alpha-2a for treatment of herpes zoster in immunosuppressed patients with cancer. AB - PURPOSE: Acyclovir and high doses of intramuscular leukocyte interferon have been shown to prevent dissemination of herpes zoster in cancer patients with localized herpes zoster. With the availability of recombinant interferon, we decided to conduct a multicenter, placebo-controlled, double-blind trial of intramuscular recombinant interferon alpha-2a to assess its efficacy and safety in the treatment of localized herpes zoster in immunosuppressed patients with cancer. PATIENTS AND METHODS: Immunosuppressed cancer patients with localized herpes zoster were randomly assigned to receive placebo, 36 X 10(6) units of recombinant interferon alpha-2a per day, or 68 X 10(6) units of recombinant interferon alpha 2a per day. Due to frequent adverse effects, the 68 X 10(6) unit dose of interferon was discontinued prior to conclusion of the trial. RESULTS: Dissemination of herpes zoster occurred in 14 of the 24 patients (58 percent) who received placebo but in only four of 24 recipients (17 percent) of 36 X 10(6) units of interferon per day (p = 0.003). Adverse effects (fever, chills, headaches, gastrointestinal irritability, fatigue, and myalgias) were more common or severe in interferon-treated patients. CONCLUSION: These results suggest that interferon modifies the severity of herpes zoster in immunosuppressed patients with cancer but is associated with frequent side effects. PMID- 3041832 TI - Southern Society for Clinical Investigation and its future viability. PMID- 3041831 TI - Presentation of the Founder's Medal of the Southern Society for Clinical Investigation to Dr. Gerald S. Berenson. PMID- 3041830 TI - Prospective study of lower respiratory tract infections in an extended-care nursing home program: potential role of oral ciprofloxacin. AB - PURPOSE: Infections of the lower respiratory tract pose an important problem in nursing homes. Despite the magnitude of this problem, few, if any, antibiotic studies have been targeted specifically at nursing home-acquired bronchopulmonary infections. Following the establishment of a teaching Extended-Care Nursing Home Program, which facilitated the early diagnosis and therapy of bronchopulmonary infections, a comparative trial of oral ciprofloxacin and intramuscular cefamandole was initiated in elderly patients with lower respiratory tract infections. In addition to assessing the relative efficacy and safety of ciprofloxacin and cefamandole, our goals were to identify problems and pitfalls associated with conducting clinical research in this nursing home setting, evaluate selected clinical and laboratory features of lower respiratory tract infection in this patient population, and measure outcomes in all study groups. PATIENTS AND METHODS: During a 20-month period, 40 patients with pneumonia and 20 patients with acute bronchitis were enrolled in this randomized study. Sixty three patients with pneumonia who were ineligible for the randomized study were also followed prospectively. The mean age of the 111 participants (123 cases) was 80.8 years; all patients had at least one chronic medical condition. RESULTS: Although Streptococcus pneumoniae was the single most common isolate, gram negative bacteria were cultured from 81 percent of the cases that yielded pathogens from a satisfactory sputum specimen. The in-hospital mortality rate was strikingly low (6.5 percent), and a large majority of patients in all study groups were discharged safely back to their nursing homes well within the Diagnosis-Related Group length of stay. CONCLUSION: Ciprofloxacin appeared to be as safe and effective as cefamandole in this nursing home program; however, additional studies are needed to determine its role in the treatment of elderly patients with bronchopulmonary infections. PMID- 3041833 TI - Evolving concepts in the treatment of acute myocardial infarction. AB - Recent studies in patients with transmural acute myocardial infarction have demonstrated that intravenous thrombolytic therapy with streptokinase or tissue plasminogen activator improves left ventricular function and reduces mortality. To accomplish this, these agents must be infused early, ie, within 3 to 4 hours of the onset of chest pain; later administration of the agents exerts no significant beneficial effect. Tissue plasminogen activator appears to be the most effective and safest of the available thrombolytic agents: its intravenous administration is followed by coronary reperfusion in about 70% of patients, and its use is not associated with allergic reactions, a systemic fibrinolytic state, or a prolonged fibrinolytic effect. Once reperfusion has been established with an intravenous thrombolytic agent, intravenous heparin is given for several days, followed by oral aspirin to prevent reocclusion. Since many of these patients have a residual high-grade coronary artery stenosis in the infarct-related artery, mechanical alleviation of the residual stenosis with angioplasty or bypass surgery is an attractive therapy 2 to 4 days after reperfusion, and preliminary data indicate that elective coronary angioplasty 3 days after thrombolytic therapy is beneficial. However, further studies are needed to assess more definitively the use of such an aggressive therapeutic strategy. PMID- 3041834 TI - Antley-Bixler syndrome from a prognostic perspective: report of a case and review of the literature. AB - The Antley-Bixler syndrome (ABS) is characterized by craniosynostosis, radiohumeral synostosis, and femoral bowing. Other findings include a trapezoid shaped head, deformed ears, severe midface hypoplasia, choanal atresia or stenosis, and long bone fractures. Most ABS cases have died in the first months of life from respiratory complications. The poor prognosis in this condition makes counseling difficult and early termination of pregnancy a consideration. The medical and surgical management information presented here can be used as a guide for counseling parents in the future. We report on a new patient with ABS who now at age 3 yr, has been followed by the medical staff of Riley Children's Hospital since birth. She has had successful medical and surgical management. Although the multisynostoses seen in this disorder is undoubtedly related to the soft tissue malformations such as choanal stenosis and midface hypoplasia, the cause remains unknown. The literature is also reviewed in this condition. PMID- 3041835 TI - Idiopathic multicentric osteolysis: report of two new cases and a review of the literature. AB - Idiopathic multicentric osteolysis is a rare skeletal disorder, usually presenting in early childhood with a clinical picture mimicking juvenile rheumatoid arthritis. Progressive destruction of the carpal and tarsal bones usually occurs and other bones may also be involved. Chronic renal failure is a frequent component of this syndrome. Mental retardation and minor facial abnormalities have been noted in some patients. We report on 2 unrelated, sporadic cases, one with facial anomalies and the other with nephropathy. Our second patient is the first black child to be diagnosed with this disease. The mode of presentation, differential diagnosis, and natural history of this disorder are briefly reviewed. PMID- 3041836 TI - The important 3:00 am blood glucose. PMID- 3041837 TI - Quick! Teach this patient about insulin. PMID- 3041839 TI - Corneal metastatic calcification in Werner's syndrome. AB - We examined twin sisters with a clinical picture typical of Werner's syndrome. Both had undergone bilateral cataract extraction, one at 39 and one at 36 years of age, and had subsequently developed bilateral corneal metastatic calcification within a period of one to two years. In one twin, this keratopathy was associated with hypercalcemia. Each of the twins underwent penetrating keratoplasty in one eye, which was complicated by recurrence of metastatic calcification in a previously normal and clear corneal graft. PMID- 3041838 TI - Echographic characteristics of benign orbital schwannomas (neurilemomas). AB - We examined two patients with orbital schwannomas (neurilemomas). The echographic findings, including a sharply outlined capsule, a well-defined central cystic space within the tumor with very low internal reflectivity surrounded by smaller cysts with variable reflectivity, slight or no compressibility, and blood flow, should help to differentiate these benign tumors from other orbital lesions. Histologic examination showed a combination of Antoni type A (dense and cellular) and Antoni type B (loose, edematous, or necrotic) patterns. PMID- 3041840 TI - A molecule resembling fibroblast growth factor in aqueous humor. PMID- 3041841 TI - Recurrent nanophthalmic uveal effusion syndrome following laser trabeculoplasty. PMID- 3041842 TI - Recurrent lacrimal abscess caused by Eikenella corrodens. PMID- 3041844 TI - The diagnosis of depression and the DSMs. AB - The classification of depression in DSM-III and DSM-III-R is radically changed from that of DSM-I and DSM-II. To understand the many changes, this paper explores early diagnostic systems, newer research studies, DSM-I, DSM-II, DSM III, and DSM-III-R. A conclusion is reached that DSM-III and DSM-III-R offer both advantages and disadvantages to DSM-I and DSM-II. These are detailed in the paper. PMID- 3041843 TI - Mast cell activation by group A streptococcal polysaccharide in the rat and its role in experimental arthritis. AB - Acute edematous responses were induced in Sprague-Dawley rats by the intravenous injection of group-specific polysaccharide (PS) isolated from group A streptococci. Thirty minutes after the intravenous injection of PS there was marked degranulation of subcutaneous and periarticular mast cells in all 4 feet, carbon particle labeling of adjacent venules, and an 8-fold increase in Evans blue dye content of the extremities. This acute reaction to PS was completely blocked by pretreatment with compound 48/80, but the polyarticular relapsing arthritis following the systemic injection of an arthropathic dose of streptococcal cell wall fragments containing large, covalently bound peptidoglycan-polysaccharide (PG-PS) was not blocked. PMID- 3041845 TI - Diagnosis of hypochondriasis: a promenade through the psychiatric nosology. AB - A new approach to the diagnosis of hypochondriasis is proposed. The hypochondriacal syndrome is first defined as a cluster of features that can be found in various mental disorders, and its differential diagnosis is then presented. Hypochondriasis is demonstrated to arise from the hypochondriacal syndrome as a distinct psychiatric entity that may for the sake of classification either belong to Anxiety Disorders and/or reflect an underlying depression, or it may be diagnosed in conjunction with one of the personality disorders or with major depression. PMID- 3041846 TI - Cognitive analysis of multiple personality disorder. AB - Multiple personality disorder is not rare, and it can be treated using the principles of cognitive therapy. Noncognitive techniques are also required. The purpose of this paper is to define the basic cognitive map of multiple personality disorder, one which recurs in the majority of cases. Multiple personality patients commonly make the classical cognitive errors such as selective abstraction and dichotomization, but they also have a set of schemata and cognitions derived from their abusive childhoods that are specific for the disorder. PMID- 3041847 TI - Narcissistic personality disorder: clinical features. AB - The author discusses the DSM-III-R diagnostic criteria of narcissistic personality disorder in the context of Kernberg's and Kohut's observations and theorizing. He also describes other clinical manifestations of this disorder often mentioned in the literature but not included in DSM-III-R, and attempts to integrate these features into a comprehensive clinical description that distinguishes (1) features that are invariably present either emanating from the grandiose self or defensively protecting or covering up the grandiose self, and (2) features that become manifest episodically in the setting of narcissistic involvements with other people. PMID- 3041848 TI - A tribute to Dr. Franz M. Enzinger. PMID- 3041849 TI - Lymphocyte predominant Hodgkin's disease nodular subtype with coexistent "large cell lymphoma". Histological progression or composite malignancy? AB - Nodular lymphocyte predominant (NLP) Hodgkin's disease (NLP HD) has been recently suggested to be of B-cell derivation on the basis of phenotypic and morphologic findings. Consistent with this view, there are sporadic case reports of coexisting NLP HD and large cell lymphoma (LCL). We describe a compilation of seven unique cases of NLP HD selected from the NIH case consultation files in which lymphohistiocytic mononuclear variant cells (L&H) became clustered into increasingly large aggregates. In areas of the same tumor mass, large confluent sheets of these cells resembled LCL. In contrast to what would be expected for LCL, all patients had localized disease clinically, and six of seven achieved long-term disease-free survival following radiation therapy or chemotherapy (two cases) for HD. None of the patients developed disseminated LCL. Also notable was a high frequency of axillary lymph node involvement (five of seven) and the high rate of occurrence in blacks (six of seven). Immunophenotypic (in two cases) and molecular genetic analysis (in one case) was suggestive of B-cell derivation for the proliferating cells. These findings also raise the possibility that some so called large cell lymphomas may actually represent histologically progressed NLP HD, and that such cases might be associated with a favorable prognosis comparable to that seen in NLP HD. PMID- 3041850 TI - Occult tumor cells in the lymph nodes of patients with pathological stage I malignant melanoma. An immunohistological study. AB - We examined 2,227 lymph nodes from 100 patients with clinical Stage I cutaneous melanoma for the presence of microscopic deposits of tumor. On examination of hematoxylin-and-eosin-stained sections, none had melanoma. Sixteen nodes from 14 patients had melanoma detectable by an antiserum to S-100 protein in a peroxidase antiperoxidase (PAP) assay. The melanomatous nature of these cells was confirmed by their reaction with the melanoma-directed monoclonal antibody NKl/C3. The incidence of occult nodal metastases was highest in patients with deeply invasive and micrometrically thick primary tumors. The incidence of occult melanoma was not increased where additional serial sections were cut and semiserial sections examined. Pitfalls in the identification of occult melanoma cells (OMC) include S 100 protein-positive interdigitating dendritic cells, capsular nevus cells, a minority of sinus "macrophages," and the Schwann cells of node-associated nerves. Thus, we conclude that the incidence of early melanoma metastases in the regional lymph nodes of patients with clinical Stage I melanoma is greater than has previously been appreciated on the basis of assessment of routine hematoxylin-and eosin-stained sections. Six of the 14 patients with OMC died of melanoma (41%), as compared to only 18 of 86 patients without OMC (21%; 0.10 greater than P greater than 0.05). PMID- 3041851 TI - Renin-producing ovarian tumor. A case report with immunohistochemical and electron-microscopic study. AB - A 39-year-old woman presented with arterial hypertension. Examination of the patient revealed elevated plasma renin activity, hyperaldosteronemia, hypokalemia, and a pelvic mass. Subsequently, an 11-cm right ovarian tumor mass with histologic features of an unusual stromal cell tumor was resected. Immunohistochemical studies demonstrated renin production by tumor cells. Organelles resembling mature renin granules were identified by electron microscopy. Although blood pressure normalized after the initial surgery, the hypertension resumed with later recurrence of the tumor. We believe the tumor originated from renin-secreting ovarian stromal cells, possibly granulosa cells. PMID- 3041852 TI - Hyperreactive malarial splenomegaly in Venezuela. AB - A cross-sectional seroepidemiological survey seeking hyperreactive malarial splenomegaly was carried out in isolated Yanomami hamlets in Amazonas Territory in Venezuela. All 110 inhabitants greater than 1 year of age were evaluated clinically and 98 were studied immunologically. The spleen index for individuals greater than 10 years of age was 44%. Only 3 patients had Plasmodium spp. on thick blood smears. All had serological evidence of infection with Plasmodium falciparum and P. vivax. Twenty-three patients were considered to show hyperreactive malarial splenomegaly. Clinical manifestations of the syndrome did not differ from those described in other parts of the world. PMID- 3041853 TI - Ultrastructural study of the effects of chloroquine and verapamil on Plasmodium falciparum. AB - Verapamil, a calcium antagonist, has recently been shown to reverse chloroquine resistance in malarial parasites in vitro. We report the first ultrastructural morphological changes associated with this phenomenon using chloroquine-sensitive and -resistant clones of Plasmodium falciparum. While the administration of 6.3 x 10(-8) M chloroquine had little morphological effect on the chloroquine-resistant strain, the combination of chloroquine and verapamil resulted in typical chloroquine-related food vacuolar swelling with increased amounts of granular matrix. Secondary morphological changes included degeneration of nuclei, mitochondria, and other organelles. These effects appeared similar to those in the chloroquine-sensitive strain of P. falciparum treated with chloroquine alone or with the chloroquine/verapamil combination. Furthermore mild food vacuolar changes were seen in a small number of parasites (from both chloroquine-sensitive and -resistant groups) exposed to high concentrations (1 x 10(-4) M) of verapamil alone. PMID- 3041854 TI - Patterns of pigment accumulation in Plasmodium falciparum trophozoites in peripheral blood samples. AB - Ninety-five samples of peripheral blood from patients with Plasmodium falciparum malaria in southwest Saudi Arabia were examined by Giemsa staining and darkfield microscopy under flow condition. Eighty-four samples contained trophozoites (ring forms) only and 11 samples contained gametocytes and trophozoites. Two patterns of pigmentation were observed in the trophozoite-containing samples: 48 (57%) contained trophozoites in which no pigment could be detected, 32 (38%) contained trophozoites with clearly detectable pigment, and 4 (5%) contained both pigmented and nonpigmented forms. Trophozoite pigmentation did not correlate with percent parasitemia or age or sex of the patients. These results indicate that microscopically observable pigment accumulation in trophozoites of P. falciparum is not required during the asexual multiplication cycle. Pigment accumulation may be triggered later in infection, perhaps as a feature of the differentiation process leading to the formation of gametocytes. PMID- 3041855 TI - Monoclonal and polyclonal antibodies both block and enhance transmission of human Plasmodium vivax malaria. AB - Antibodies against gametes of the malarial parasite inhibit the development of the parasite in the mosquito and curtail the transmission of malaria. We now report that a monoclonal antibody against gametes of the human malaria pathogen Plasmodium vivax and antibodies induced during natural infections of P. vivax in humans which suppress infectivity of the parasites to the vector at high concentrations can, at lower concentrations, have the opposite effect and enhance the level of malaria infection in the mosquitoes. Infectivity enhancing effects of up to 12-fold were demonstrated when a transmission blocking monoclonal antibody and immune human sera were diluted, in some undiluted immune human sera, and in the sera of vivax malaria patients during convalescence after drug cure. PMID- 3041856 TI - Human cerebral malaria. AB - Possible factors contributing to the development of cerebral malaria were discussed based on pathological changes in Burmese patients who died of cerebral malaria. Blockage of cerebral capillaries by Plasmodium falciparum infected erythrocytes appeared to be the principal cause of cerebral malaria. From electron microscopic results, it was concluded that knobs on infected erythrocytes acted as focal junctions which mediated adhesion to endothelial cells. The knobs are, therefore, important contributors to the blockage of the capillary lumen and ensuing pathological changes in cerebral tissues. Host cell molecules such as OKM5 and thrombospondin may function as endothelial cell surface receptors for the attachment of knobs of P. falciparum infected erythrocytes. Immunological events might also play a role in the pathogenesis of cerebral malaria. This was suggested by the presence of IgG, IgM, P. falciparum antigens, and knob proteins in the cerebral capillaries of the people with cerebral malaria. It will be important to assess the candidate malaria vaccines now in development not only for their efficacy in reducing parasitemia but for effects they may have on the sequestration of infected erythrocytes in the brain. PMID- 3041857 TI - High salt lysates: a simple method to store blood samples without refrigeration for subsequent use with DNA probes. AB - Blood specimens to be tested for the presence of Plasmodium falciparum using specific DNA probes can be stored as high salt lysates (HSL) without refrigeration. The lysates are prepared from 100 microliter blood samples by a simple 3-step procedure using 2 volumes of H2O to lyse the erythrocytes (step I), 1 volume of a detergent/EDTA mix to lyse the parasites (step II), followed by the addition of 1 volume cesium trifluoroacetate (CsTFA) (step III). The parasite DNA was found to be undegraded, as shown by the unaltered pattern of repetitive sequences obtained after storage of up to 1 month at 37 degrees C, due to the inhibition of DNA degrading enzymes by the cesium salt. The bulk of protein can be removed from the samples by a 1-step precipitation. The addition of 0.3 volumes of a mixture of ethanol: chloroform: isoamyl alcohol (2.5:1:0.04 v/v) precipitates greater than 90% of the proteins from the lysates, leaving greater than 86% of the parasite DNA in the supernatant. The reduced protein content of the samples, when applied to solid supports, results in an increased signal: background ratio on autoradiograms. PMID- 3041858 TI - IgE antibody production and cutaneous anaphylactic reactions in rats infected with Clonorchis sinensis. AB - Rats were infected orally with 50 or 100 metacercariae of Clonorchis sinensis. Anti-Clonorchis IgE antibody, determined by passive cutaneous anaphylaxis, in the serum of infected rats appeared 30-40 days after infection and persisted for at least 120 days. Total amount of IgE fixed on the mast cells, detected by reverse passive cutaneous anaphylaxis with anti-rat IgE, was significantly more in the infected rats than in uninfected rats. Worm burden at 290 days post-infection did not correlate to anti-Clonorchis IgE antibody titers, reverse passive cutaneous anaphylaxis titers, or anti-Clonorchis anaphylactic antibody on the mast cells, determined by active cutaneous anaphylaxis. Active cutaneous anaphylaxis titers interrelated to reverse passive cutaneous anaphylaxis titers, suggesting a correlation between the amount of anti-Clonorchis anaphylactic antibodies and total IgE on the mast cells. Sensitivity of passive cutaneous anaphylaxis with anti-BSA IgE antibody was significantly suppressed in Clonorchis infected rats. Moreover, anti-BSA IgE titers in the infected rats were inversely related to reverse passive cutaneous anaphylaxis titers, indicating that increase of IgE antibody on the mast cells by Clonorchis infection interfered with sensitization of anti-BSA IgE antibody. PMID- 3041859 TI - Foods as a source of enteropathogens causing childhood diarrhea in Thailand. AB - Foods obtained in markets in Bangkok were cultured for bacterial enteric pathogens and examined for their similarity to strains isolated from children under 5 years of age in Bangkok in 1986. Salmonella was isolated from 17%, Campylobacter from 12%, and enterotoxigenic Escherichia coli (ETEC) from 3% of 510 foods examined. Campylobacter was isolated from 13.5%, ETEC from 13%, and Salmonella from 12% of 1,230 children under 5 years of age with diarrhea. Eighty eight percent of children infected with Salmonella were infected with serotypes isolated from foods of animal origin. Six percent of children with Salmonella were infected with the same serotype containing plasmids with identical endonuclease restriction patterns as isolates from food. Eighty-seven percent of children with Campylobacter were infected with the same serotypes and biotypes found in food of animal origin. Thirty-one percent of heat-labile enterotoxin (LT) producing ETEC from foods containing genes coding for LT II, but LT II ETEC was not isolated from children. Twenty-one percent of ETEC isolated from foods vs. 53% isolated from children were resistant to 2 or more antibiotics (P less than 0.01). Salmonella and Campylobacter, but not ETEC, isolated from foods were similar to strains isolated from children. Foods of animal origin are an important source of Salmonella and Campylobacter in Thailand. PMID- 3041860 TI - Aortobrachiocephalic reconstruction. AB - Aortobrachiocephalic reconstruction was studied in 26 patients. Nine had suffered a previous stroke, and seven had residual deficity. A previous carotid endarterectomy or carotid-to-subclavian bypass had been performed in eight patients. The ascending aorta was the proximal anastomotic site in all cases. Bypass grafting to a single distal site was performed in 11 patients (42 percent), to 2 distal sites in 14 patients (54 percent) and to 3 distal sites in 1 patient (4 percent). The carotid artery, innominate arteries, or both were involved in all reconstructions. Concomitant carotid or subclavian endarterectomy was performed in 14 patients, 1 of whom also had a coronary bypass at the same time. Two patients (7 percent) died in the postoperative period. Twenty-three of 24 survivors (96 percent) had relief of symptoms after operation. Only one patient had worsening of symptoms postoperatively, which was secondary to intracerebral hemorrhage. Results of this study indicate that direct revascularization of the arch branches can be carried out with minimal morbidity and mortality. Relief of presenting symptoms is to be expected, and long-term results suggest that the operation is durable. PMID- 3041861 TI - Origin of deep vein thrombi in an ambulatory population. AB - Eighty symptomatic ambulatory outpatients with acute deep vein thrombosis were evaluated with ascending contrast venography and ultrasonic imaging to determine the origin and distribution of thrombosis and to analyze clinical risk factors. Isolated calf vein thrombosis was present in 42.5 percent of the limbs, combined calf and proximal deep vein thrombosis in 47.5 percent, and isolated proximal thrombosis in 10 percent of the limbs. Discontinuity of thrombus was present in 55 percent, whereas 45 percent exhibited a continuous column of thrombus. The results of this study indicate that in the ambulatory outpatient population, acute deep vein thrombosis begins segmentally in the calf and proximal vessels and frequently coalesces into a continuous column of thrombus over several days. We believe that all cases of acute deep vein thrombosis should be treated and patients with evidence of previous acute deep vein thrombosis should be closely monitored for possible recurrences. PMID- 3041862 TI - Quantification of human spiral ganglion cells by serial section reconstruction and segmental density estimates. AB - Spiral ganglion cell populations were determined for six normal adults using two methods, 1) graphic reconstruction to calculate total segmental spiral ganglion counts, and 2) measurement of spiral ganglion cell density for given cochlear segments. Such data can be used to evaluate possible loss of spiral ganglion cell populations in pathologic human specimens. The method of segmental density calculation will be particularly useful when serial sections of the entire spiral ganglion are not available. PMID- 3041863 TI - Applications of hyperbaric oxygen for the otolaryngologist--head and neck surgeon. AB - In the last 10 years, the use of hyperbaric oxygen (HBO) in clinical medicine has increased dramatically. Despite its controversial beginnings, it has won acceptance as a treatment for specific diseases in both medical and surgical patients. The purpose of this article is to familiarize the otolaryngologist-head and neck surgeon with the currently accepted applications of HBO and to present some promising investigational uses of this therapy. PMID- 3041864 TI - Experimental otitis media with effusion induced by lipopolysaccharide from Klebsiella pneumoniae: mucociliary pathology of the middle ear. AB - We inoculated 100 micrograms/ml of lipopolysaccharide (LPS) from Klebsiella pneumoniae into the tympanic cavity of guinea pigs and examined the mucociliary pathology in the middle ear. Serous effusion was observed in the tympanic cavity of every animal on the first, third, and seventh day following the procedure, but the volume of the effusion had decreased to 0.2 ml on day 7. By that time, the ciliary activity in the opening to the eustachian tube within the middle ear had recovered to some extent, but in the middle ear distal to the opening no recovery was apparent. Our results show that cilia close to the eustachian tube play a more significant role in middle ear clearance than those in the middle ear distal to the tube. Compared with our previous study using 10 micron/ml of LPS, this study also demonstrates that inoculations with a higher concentration of LPS induces longer-term middle ear effusions. PMID- 3041865 TI - Plasma insulin response to oral glucose load in Meniere's disease. PMID- 3041867 TI - [Prostaglandins as mediators of immunity. The importance of prostaglandins in human reproduction]. PMID- 3041866 TI - [Diagnosis and echographic control of the treatment of uterine myoma in girls and young women]. PMID- 3041868 TI - [The complex diagnosis of ovulation]. PMID- 3041869 TI - [Fibrocystic mastopathy (its pathogenesis, clinical picture and therapy)]. PMID- 3041870 TI - History of medicine in Alaska. Robert Bocock Wilkins, M.D. PMID- 3041871 TI - Smallpox decimates the Tlingit (1787). PMID- 3041872 TI - Early Alaska mental health. PMID- 3041873 TI - Overtransfusion as a possible cause of split skin graft loss. AB - A child who underwent burns surgery received excessive transfusion of red blood cells during operation and subsequently suffered severe skin graft and donor site loss. The possible causes are discussed and hyperviscosity is suggested to be the most probable. PMID- 3041874 TI - [Hemodynamics in donor plasmapheresis]. AB - Several studies have demonstrated that preoperative withdrawal and storage of autologous plasma as fresh frozen plasma is effective in blood conservation. For that purpose patients with elective surgery (orthopaedic surgery, open heart surgery, neurosurgery and others) have to undergo donor plasmapheresis without staying in the hospital. Depending upon the need the procedure can be performed several times preoperatively, taking about 900 ml in a normal weighting subject at once. The collection of autologous plasma should be finished at least 14 days before surgery. In order to investigate the haemodynamic effects of donor plasmapheresis 30 patients scheduled for coronary bypass surgery were devided into two groups. 15 patients underwent plasmapheresis (10 ml plasma/kgbw) by one needle-technique using a Haemonetics seperator (PCS) after premedication but before onset of anaesthesia. Blood withdrawal was performed with 0.5 ml/kgbw x min. Another 15 patients, serving as control had no plasma withdrawal and were measured at identical times as the other group. Both groups had an identical fluid replacement with 500 ml Ringer's solution during the investigation period. Plasma withdrawn was not substituted by colloidal solution (simulating the situation when plasmapheresis is performed at the outpatient). Haemodynamic measurements (both groups) included heart rate, arterial blood pressure, right- and left-atrial pressure, systemic- and pulmonary-vascular resistance and cardiac output. There were no relevant effects of plasmapheresis on haemodynamic function during and after the investigation period in that patients: neither heart rate, blood pressure or vascular resistance changed significantly nor did pre- and afterload or cardiac index. No differences to the group without plasmapheresis could be observed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3041875 TI - [Anniversary calendar of theoretical and experimental morphology 1988]. PMID- 3041877 TI - Hereditary angioedema: clinical and biochemical heterogeneity. AB - Hereditary angioedema is a well defined entity for which we now have effective therapy. It is a condition however that can have different clinical presentations which require individualized therapeutic approaches. The recent advances into the molecular biology of this condition should provide further insights for its effective management. PMID- 3041876 TI - Laminar flow air cleaner bed attachment: a controlled trial. AB - Thirteen house dust mite-allergic young asthmatics were entered into a double blind, crossover clinical trial to compare "in use" with "non-use" of a laminar flow air cleaner bed attachment. The study design encouraged maintenance of symptom control by adjustment of the dose of medications used. There was a significant reduction in the amount of medications required by the patients during the trial period that the air cleaner was in use. Histamine airway responsiveness was significantly decreased for a group of eight of the subjects tested at the end of a 3-month open trial. PMID- 3041878 TI - Chronic cough in a 61-year-old female. PMID- 3041879 TI - Solid-phase enzyme immunoassay for anti-mite IgE and IgG antibodies in allergic patients. AB - Solid-phase enzyme immunoassays for the measurement of anti-mite IgE and IgG antibodies in the sera of mite-allergic patients were developed. In both assays, microtiter plates coated with the crude extract of Dermatophagoides farinae were used. The solid-phase enzyme immunoassay detected mite-specific IgE and IgG antibodies in sera of 18 and 15 out of 20 allergic patients, respectively. The correlation coefficient between the two antibody titers in patients' sera was 0.74, which was statistically significant (P less than .01). Both of the assays require no special facilities for radioactive materials and are considered to be useful in practice for the diagnosis of allergy to house dust mites by combining the results of the two assays. PMID- 3041880 TI - Strains of Escherichia coli associated with urogenital disease in dogs and cats. AB - Selected strains of Escherichia coli associated with urogenital disease in dogs and cats were evaluated for 3 virulence factors associated with human uropathogenic strains. Urogenital strains of E coli from dogs and cats had high prevalence of alpha hemolysin and were clustered in 5 to 10 somatic serogroups, attributes also shared by human uropathogenic strains. However, the canine and feline urogenital strains failed to have increased prevalence of mannose resistant hemagglutination, as has been reported for human uropathogenic strains. PMID- 3041881 TI - Clinical and serologic evaluations of induced Borrelia burgdorferi infection in dogs. AB - Adult Beagles were used to evaluate clinical signs and serologic response after inoculation with, or exposure to, Borrelia burgdorferi. An indirect immunofluorescent assay (IFA) and 2 ELISA were used to monitor the serologic response to B burgdorferi. Feeding infected ticks on 4 dogs (group 1) failed to cause seroconversion, and SC inoculation with 500 organisms caused minimal seroconversion in 2 of 4 dogs (group 2). At 56 days, approximately 3.01 X 10(8) B burgdorferi organisms were injected IV into group-1 dogs, and intraperitoneally into group-2 dogs. A control group of 4 dogs (group 3) had noninfected ticks feed on them, and then were given IV injection of physiologic saline solution. Increases in immunoglobulin M (IgM) titers were detected in 2 of 4 group-2 dogs approximately 7 days after the initial exposure. These titers returned to negligible values 20 days later. Immunoglobulin G titers increased approximately 10 days after the initial exposure and were mildly increased 56 days later, when dogs were exposed a second time. Both the IV and intraperitoneal injections (second exposures) resulted in increased IgM titers, which in both groups eventually returned to preexposure values after approximately 2 months. Immunoglobulin G titers increased within a week after the second exposure, and in 3 dogs monitored for 8 months, returned to negligible values after the 8-month period. One control dog had a slightly increased IgG titer 24 days after the second inoculation. The possibility of urine transmission is suggested. Clinical status, hemograms, serum biochemical profiles, ECG and results of urinalyses remained normal throughout the study.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3041882 TI - Production and characterization of monoclonal antibodies to bovine beta 2 microglobulin. AB - In an attempt to isolate monoclonal antibodies specific for bovine lymphocytes, spleen cells from mice immunized with bovine lymphocytes were fused with the mouse myeloma cell line SP-2/0. The resulting hybridoma cell lines were tested for reactivity with bovine lymphocytes, polymorphonuclear neutrophils, RBC, gamma globulin, kappa-casein, beta-casein, alpha-S1-casein, and beta 2-microglobulin (beta 2m) and with beta 2m from rabbits, goats, and human beings. None of the clones secreted anti-bovine lymphocyte-specific antibody. However, 4 secreted monoclonal antibodies to bovine beta 2m. They also reacted with beta 2m from rabbit, goat, and human being. One monoclonal antibody also was found to be reactive with bovine immunoglobulin. Monoclonal antibodies to beta 2m could serve as a tool to (1) explore the homology of the beta 2m molecule among various species, (2) examine the relationship of beta 2 m with the constant region of the immunoglobulin molecule, (3) quantitate bovine beta 2m in various body fluids and major histocompatibility antigens on cell surfaces, (4) help characterize those antigens in cattle, and (5) be used for tissue typing of those antigens. PMID- 3041884 TI - Histologic features of the healing of bone graft donor sites in dogs. AB - Healing of cancellous bone graft donor sites in the proximal tibial metaphysis of 12 healthy adult dogs was studied histologically. Cancellous bone was curetted from the metaphysis of the proximal end of the tibia, via a 1-cm diameter circular opening in the medial cortex. A hematoma and fibrovascular tissue filled the bone defect at 2 weeks. At 4 and 8 weeks, endosteal callus, composed initially of cartilage and woven bone and later of lamellar bone, filled the marrow cavity. At 12 weeks, the normal structural arrangement of lamellar bone and hematopoietic marrow was reestablished in the marrow cavity. The medial cortex defect was filled only with lamellar trabecular bone. It was concluded that, in adult dogs, a second cancellous bone graft could be collected from the proximal portion of the tibial metaphysis 12 weeks or more after an initial collection. PMID- 3041883 TI - Biomechanical properties of canine cortical bone allografts: effects of preparation and storage. AB - The effects of various preparation and storage procedures and of different storage times on structural properties of canine cortical bone allografts were determined by evaluation of the compressive load to failure of a whole diaphyseal segment, the ability of a screw to resist being pulled from a cortical segment, and the torque required to strip the threads of a screw hole in a cortical segment. Preparation and storage procedures evaluated were sterile collection and storage at -20 C; ethylene oxide sterilization and storage at room temperature (22 C); chemical sterilization (methanol and chloroform, then iodoacetic acid) and storage at -20 C; and chemical sterilization, partial decalcification, and storage at -20 C. Storage times were 1, 16, and 32 weeks for each procedure. After 1 week of storage, aseptically collected frozen bone and ethylene oxide sterilized bone had an increase, compared with matched controls, in load to failure in compression, but pullout load or screw-stripping torque did not change. Chemically sterilized bone had not changed after 1 week of storage, whereas chemically sterilized and partially decalcified bone had a 40% to 60% decrease in compressive load to failure, pullout load, and screw-stripping torque. Chemically sterilized and partially decalcified bone remained weak after 16 and 32 weeks of storage. Significant structural alterations were not detected in aseptically collected bone after 16 or 32 weeks of storage. Ethylene oxide sterilized bone had a reduced pullout load after 32 weeks of storage. Chemically sterilized bone had significantly reduced compressive load to failure and pullout load after 16 and 32 weeks of storage.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3041885 TI - [Neurologic aspects of Ito's hypomelanosis]. AB - Hypomelanosis of Ito (incontinentia pigmenti achromians) is a neurocutaneous syndrome consisting of bizarre, patterned, macular hypopigmentation over variable portions of the body surface. Associated defects in other systems occur, commonly in the central nervous system, in a significant percentage of affected individuals. Six children affected by hypomelanosis of Ito are presented. Similarities and differences between hypomelanosis and incontinentia pigmenti and systematized achromic nevus are discussed. PMID- 3041887 TI - [Anterior sacral meningocele]. PMID- 3041886 TI - [Acquired immunodeficiency syndrome in childhood. Apropos of a case: management, complications and necropsy findings]. PMID- 3041888 TI - [Cholestasis as the first sign of cystic fibrosis of the pancreas]. PMID- 3041889 TI - [Cervical teratoma in a newborn infant. Prenatal diagnosis. Surgical treatment. Posterior evaluation]. PMID- 3041890 TI - [Holoprosencephaly in a child of a diabetic mother]. PMID- 3041891 TI - [Castleman's disease in a 2-year-old child]. PMID- 3041892 TI - The case of the disappearing and reappearing intraocular lens. AB - A 75-year-old woman who underwent implantation of an anterior-chamber intraocular lens was noted to have the lens disappear from the anterior chamber four days after surgery and reappear on the eleventh postoperative day. Ultrasound documented the position of the lens as apparently lying posterior to the sclera and anterior to the ciliary body. Extrusion through a cyclodialysis cleft superonasally was the apparent mechanism of exit and reentry. Pseudophakic dislocation is becoming increasingly common. PMID- 3041893 TI - Third outbreak of trichinellosis caused by consumption of horse meat in Italy. AB - The clinical and epidemiological aspects of the largest Italian outbreak of human trichinellosis which occurred in Northern Italy in August 1986 are reported. About 300 people who ate raw horse meat were involved. The clinical and biological aspects of 161 patients were studied. None died. The causal parasite, isolated from a patient, was identified through isoenzymatic typing as Trichinella nelsoni. Fever, oedema, myalgia and headache were the most common signs and symptoms. The Fluorescent Antibody Test (FAT) proved positive for 96 percent of the patients showing clinical manifestations. Essential laboratory data are reported. PMID- 3041894 TI - [The rabbit, experimental host of Pneumocystis carinii]. AB - Pneumocystis carinii can be collected from experimentally immunodepressed rats. However, development of experimental pneumocystosis in this host requires corticoid-treatment during 8 to 12 weeks while P. carinii can be obtained from immunodepressed rabbits after 2 to 4 weeks. In 400 to 650 g body weight-white rabbits normally fed, two immunodepression protocols were used: (a) daily subcutaneous injections of hydrocortisone acetate (10 mg/kg body weight) and G penicillin (25,000 IU)-streptomycin (3 mg); (b) continuous oral administration of prednisolone (20 mg/l/day) and amoxicillin (25 mg/l/day) both diluted in drinking water. Between 55 and 78% of rabbits treated by any immunodepression protocol died within 5-20 days after a terminal important diarrhoea. Seventy and seventy seven per cent of the young rabbits respectively treated by protocols (a) or (b), showed the presence of Pneumocystis by the direct microscopic examination, but all of them were found positive after lung concentration. Fifty-five per cent of the older rabbits (950-1,400 g) were positive by pulmonary smear examination when treated with protocol (a). Pneumocystis obtained were good antigens for indirect immunofluorescence and immunoblot assays using human and rabbit sera. PMID- 3041895 TI - [Infantile acropustulosis]. PMID- 3041896 TI - [Alopecia areata in children]. PMID- 3041898 TI - [Dysplastic nevus syndrome]. PMID- 3041897 TI - [Self-limiting Hashimoto-Pritzker histiocytosis. Review of the literature apropos of 2 case reports]. PMID- 3041899 TI - Pathogenesis and biology of anoplocephaline cestodes of domestic animals. PMID- 3041900 TI - [Epidemiology of diarrhea caused by Escherichia coli and rotavirus in calves and lambs in Morocco]. AB - An epidemiological survey on E coli and rotavirus associated diarrheas in one to twenty five days old calves and lambs was made in three regions: Rabat-Kenitra, Marrakech and Agadir. Isolated E coli K99 stains have been studied of a biochemical, serotypical (O antigen) and antibiotypical point of view. The identification of rotavirus was made by ELISA test. Persistence of K99 antigen and heat stable toxin A was examined after a conservation of 5 weeks at - 18 degrees C. The frequency of E coli K99 or rotavirus associated diarrheas is respectively 26.9% and 29.7% in calf, 10% and 30% in lamb. This incidence considerably decreases from the 20th day in calf and from the 11th day in lamb. It must be observed that 34.8% of cases of diarrheas in calf and 55% in lamb cannot be ascribed to investigated agents. Only 12 out of 42 E coli K99 strains belong to serogroups O101, O8 and O9. Preservation of strains to - 18 degrees C comes with the loss of K99 antigen. These strains are not toxinogens. Among the strains having kept this antigen, 29% are toxinogens. Surveyings of antibiotics resistance was discussed. PMID- 3041901 TI - When and how should we measure plasma ammonia? AB - Hyperammonaemia is associated with a high morbidity and mortality. It is important to diagnose as it is often treatable and perhaps, most importantly, may be genetically determined. Measurement of plasma ammonia is rarely necessary in adult medicine. In paediatrics, and particularly in the neonatal period, it is an important investigation in the diagnosis and often in the subsequent management of several inherited metabolic disorders. Patients with these disorders, particularly neonates, will deteriorate over a period of hours and investigation cannot wait. In these situations a plasma ammonia together with other investigations for metabolic disorders must be available urgently and are sometimes necessary out of normal laboratory hours. Interpretation must take into account the age and maturity of the child as well as the clinical state and results of other investigations. If treatment is initiated, frequent monitoring of plasma ammonia may be required. Plasma ammonia can be reliably and conveniently measured using a specific ion-selective electrode system or an automated enzyme method. Screening using a microdiffusion method is not a satisfactory alternative to a quantitative assay. Plasma ammonia is no longer solely the province of the specialised paediatric laboratory, but should be part of the repertoire of every laboratory supporting neonatal or paediatric units. The threshold for accepting requests should be lower than at present if we are to prevent misdiagnoses. PMID- 3041902 TI - The clinical value of ionised calcium assays. PMID- 3041903 TI - Essential considerations in the provision of near-patient testing facilities. AB - (1) Near-patient testing (NPT) is both practicable and in some situations desirable. (2) Like all new technologies its apparent simplicity often belies its complexity and masks the need for attention to detail in order to achieve optimum effects and avoid disasters. (3) Though technically unskilled individuals are capable of using it, they must undergo training in the elements of safety, sample collection, quality control, quantitation and documentation before being authorized to provide analytical services for patients. (4) Health Authorities should be encouraged to adopt a policy of integration of NPT and clinical laboratory services in order to reduce unplanned use and abuse of NPT facilities which is not only wasteful and divisive, but also dangerous. This has recently been emphasized in the United Kingdom by the release of a Hazard Notice (HN (Hazard) (87) 13) by the Department of Health and Social Security. The experienced clinical biochemist will, by virtue of training and experience, usually be the most suitable person in a Health District to advise on many of the issues involved in NPT. It should therefore, be the responsibility of laboratory staff to work with management, clinical, and nursing staff to: choose appropriate sites for the various levels of service to be provided to meet a clinical need; select the equipment and reagents to be employed; provide training in the use of the apparatus, quality control and safety; authorize accredited users and oversee the quality assurance programme. (5) Laboratory staff will need to involve themselves closely in the financial implications of NPT both for the laboratory's benefit and that of clinical practice within the hospital and/or community as a whole. PMID- 3041904 TI - An enhanced chemiluminescence enzyme immunoassay for serum oestradiol. AB - Oestradiol in serum was determined with a simple enhanced chemiluminescent enzyme immunoassay. The assay is based on oestradiol labelled with horseradish peroxidase and the IgG fraction of an oestradiol antiserum coated on a black polystyrene microtitre plate. The enzyme activity of bound label was determined using a p-hydroxycinnamic acid-enhanced chemiluminescent reaction. The assay was sensitive (1.8 fmol/well), precise (intra- and inter-assay CV 4-10% and 8-12%, respectively for sample concentration in the range 122-1330 pmol/L) and showed good agreement with conventional radioimmunoassays (r = 0.99). PMID- 3041905 TI - An automated solid-phase 17 alpha-hydroxyprogesterone ELISA method using a microtitre plate. AB - We report an automated ELISA method for the measurement of 17 alpha hydroxyprogesterone in plasma samples. A rabbit antiserum raised against 17 alpha hydroxyprogesterone-3-(O-carboxymethyl)oxime-bovine serum albumin is used for the assay and a homologous competitor 17 alpha-hydroxyprogesterone-3-(O carboxymethyl)oxime-bovine thyroglobulin is coated onto a microtitre plate. A goat anti-rabbit IgG-horse radish peroxidase is used as a probe for this solid phase assay. The assay exhibits good sensitivity, precision and accuracy. The method is used routinely for the management of patients with congenital adrenal hyperplasia. PMID- 3041906 TI - [Gastro-esophageal reflux. Surgical treatment]. PMID- 3041908 TI - [The 48,XXXX syndrome: study of psychomotor development from birth to 11 years of age and review of the literature]. AB - A child with four X chromosomes is described. This case and the literature review allow to underline the mental retardation and some other "major" but inconstant signs that are extremely helpful for the early clinical diagnosis. They are hypertelorism, epicanthal fold and genital anomalies. The mental evolution is assessed on an eleven year period. The bad results concern particularly the child's use of language and the complicated works. They become worse with time. The additional X chromosomes Mary Lyon inactivation, perhaps incomplete, is discussed because its determinism. Enzymatic measuring out is our approach to this problem. PMID- 3041907 TI - [Ability of cholestyramine to bind Escherichia coli and Vibrio cholerae toxins]. AB - Cholestyramine treatment of new born's infectious diarrhea has been shown to be effective. The study of in vitro binding of bacterial toxins from Vibrio Cholerae and from three strains of Escherichia coli suggest an ionic adsorption of the four toxins to this anion-exchange resin. This immediate binding is effective at the pH of intestinal fluid, therefore the protection of the enterocytes from the biological action of the toxins result of the toxins sequestering effect of the Cholestyramine. PMID- 3041909 TI - [Mechanism of renal allograft rejection]. AB - Renal allograft rejection, in an immunocompetent subject, results from a cellular and humoral interaction. Lymphokines, which are immunity mediators (particularly interleukin 1 and 2, and gamma interferon) modulate these different cellular populations, among which T lymphocytes play a major role. The current knowledge of rejection mechanisms allows us to evaluate the principal targets of immunosuppressive drugs used to treat allograft rejection. PMID- 3041910 TI - [Organ procurement. The Clermont-Ferrand experience]. AB - The authors report the experience of a new kidney collection centre organised at Clermont-Ferrand, which rapidly became involved in the collection of multiple organs. The difficulties encountered during the collection of organs from the first 30 cadavres are presented: 19 kidneys were not grafted, sometimes for anatomical reasons, but especially because of surgical reasons (during the following period, this number fell to 3 for a series of 21 cadavres). 13 hearts and 2 livers were also removed. In conclusion, the authors stress that such organ collection centres can only be created when transplantations are also performed, because of inevitable psychological reasons affecting both the health care team as well as the population of the region. PMID- 3041911 TI - Protein phosphorylation and the respiratory burst. AB - The exposure of 32P-loaded neutrophils to any of a variety of activating agents induces changes in the levels of phosphorylation of a large number of phosphoproteins. The uptake of phosphate by one set of phosphoproteins in particular, a family whose members migrate at Mr 48K with near neutral pI values, appears to be closely related to the activation of the respiratory burst oxidase, the O2--producing enzyme of phagocytes that is responsible for the generation of microbicidal oxidants by these cells. Evidence for the relationship between the phosphorylation of these proteins and the activation of the respiratory burst oxidase has been furnished by kinetic studies as well as by studies on protein phosphorylation in neutrophils from patients with chronic granulomatous disease, a group of inherited disorders affecting this oxidase. The details of this relationship are obscure, although the evidence suggests that these phosphoproteins act in substoichiometric amounts with respect to the oxidase. PMID- 3041913 TI - Subcellular distribution and properties of carbonyl reductase in guinea pig lung. AB - On subcellular fractionation, carbonyl reductase (EC 1.1.1.184) activity in guinea pig lung was found in the mitochondrial, microsomal, and cytosolic fractions; the specific activity in the mitochondrial fraction was more than five times higher than those in the microsomal and cytosolic fractions. Further separation of the mitochondrial fraction on a sucrose gradient revealed that about half of the reductase activity is localized in mitochondria and one-third in a peroxidase-rich fraction. Although carbonyl reductase in both the mitochondrial and microsomal fractions was solubilized effectively by mixing with 1% Triton X-100 and 1 M KCl, the enzyme activity in the mitochondrial fraction was more highly enhanced by the solubilization than was that in the microsomal fraction. Carbonyl reductases were purified to homogeneity from the mitochondrial, microsomal, and cytosolic fractions. The three enzymes were almost identical in catalytic, structural, and immunological properties. Carbonyl reductase, synthesized in a rabbit reticulocyte lysate cell-free system, was apparently the same in molecular size as the subunit of the mature enzyme purified from cytosol. These results indicate that the same enzyme species is localized in the three different subcellular compartments of lung. PMID- 3041912 TI - Biochemical and immunological demonstration of prostaglandin D2, E2, and F2 alpha formation from prostaglandin H2 by various rat glutathione S-transferase isozymes. AB - Glutathione S-transferase isozymes purified from normal rat liver (1-1, 1-2, 2-2, 3-3, 3-4, and 4-4), liver with hyperplastic nodules (7-7), brain (Yn1Yn1), and testis (Yn1Yn2) all had prostaglandin H2-converting activity. The prostaglandin H2 E-isomerase activity was high in 1-1 (1400 nmol/min/mg protein), 1-2 (1170), and 2-2 (420), moderate in 3-3, 3-4, 4-4, Yn1Yn1, and Yn1Yn2 (52-100), and weak but significant in 7-7 (33). The prostaglandin H2 D-isomerase activity was relatively high in 1-1 (170) and 1-2 (200), moderate in 2-2 (60) and Yn1Yn2 (43), and weak but marked in 3-3 (16), 4-4 (16), and 7-7 (14). The prostaglandin H2 F reductase activity was remarkable in 1-1 (1250), 1-2 (920), and 2-2 (390), and weakly detected in 3-3 (24), 4-4 (28), and 7-7 (14). Glutathione was absolutely required for these prostaglandin H2-converting reactions, and its stoichiometric consumption was associated with F-reductase activity but not E- and D-isomerase activities. The Km values for glutathione and prostaglandin H2 were about 200 and 10-40 microM, respectively. By immunoabsorption analyses with various antibodies specific for each isozyme, we examined its contribution to the formation of prostaglandins D2, E2, and F2 alpha from prostaglandin H2 in 100,000g supernatants of rat liver, kidney, and testis. In the liver, about 90% of the F reductase activity (9.8 nmol/min/mg protein) was shown to be catalyzed by the 1-2 group of isozymes. The E-isomerase activity (16.5) was catalyzed about 60 and 40% by the 1-2 and 3-4 groups, respectively; and the D-isomerase activity (3.7) was catalyzed by the 1-2 group (50%) and the 3-4 group and Yn1Yn2 (15-25%). In the kidney, the E-isomerase activity (9.4) was catalyzed by 1-1, 1-2 (40%), 2-2, 3-4 group, and 7-7 (10-20%). The F-reductase activity (3.3) was mostly catalyzed by the 1-2 group (75%). In the testis, the E-isomerase activity (3.9) was catalyzed by the 1-2 group (20-30%), the 3-4 group, and Yn1Yn2 (30-60%). PMID- 3041914 TI - Incorporation of myristate and palmitate into the sheep reticulocyte transferrin receptor: evidence for identical sites of labeling. AB - The ability of sheep reticulocytes and plasma membranes isolated from them to incorporate fatty acids into the transferrin receptor has been examined using both [3H]palmitate and [3H]myristate. Both fatty acids, when incorporated into the transferrin receptor, can be released by treating the protein with 1 M hydroxylamine at pH 7.0. After treatment of the 3H-acylated receptor with borohydride, an 3H-labeled alcohol is released, suggesting that the receptor bound fatty acid is in thioester linkage. With both [3H]myristate and [3H]palmitate, Cleveland maps from immunoprecipitates of the transferrin receptor labeled in intact cells and isolated membranes show that identical peptides are labeled. No evidence was obtained for qualitatively different labeling with the two fatty acids. In intact reticulocytes, incorporation of [3H]palmitate into the transferrin receptor is approximately 3.5 times greater than the incorporation of [3H]myristate from equivalent concentrations of the labeled fatty acids. However, in isolated reticulocyte plasma membranes, there is much less difference between palmitate and myristate incorporation (with ATP) or between their acyl-CoA derivatives. The reason for the discrepancy between cells and membranes is unknown but may be due to the presence in intact cells of more than one enzyme for activating the fatty acids. Acylation of the receptor in isolated plasma membranes is fourfold greater with the CoA derivatives than with the free fatty acids. The fatty acid activating enzyme(s) as well as the acyltransferase(s) appear to be membrane bound in reticulocytes. PMID- 3041915 TI - Vancomycin-induced linear IgA bullous dermatosis. PMID- 3041917 TI - Sinus histiocytosis with massive lymphadenopathy. Current status and future directions. PMID- 3041916 TI - Autosomal recessive transmission of neuroectodermal syndrome. PMID- 3041918 TI - Atypical pigmented penile macules. AB - We describe three patients with pigmentary lesions of the penis that simulated malignant melanoma. The pathologic features were unimpressive, although one lesion may have represented a melanoma in evolution. The biologic potential of such lesions and their appropriate therapy remain to be defined. PMID- 3041919 TI - [Anatomy and development of the testicular venous system]. PMID- 3041920 TI - [Primary and secondary megaureter. Therapeutic considerations]. PMID- 3041921 TI - [Penile curvature without hypospadias: surgical treatment]. PMID- 3041922 TI - [Fibrous pseudotumor of the spermatic cord. Morphological and immunohistochemical study of a case and review of the literature]. PMID- 3041923 TI - Immunisation of patients with rheumatoid arthritis and systemic lupus erythematosus. PMID- 3041924 TI - A double blind multicentre study of OM-8980 and auranofin in rheumatoid arthritis. AB - The therapeutic efficacy of the immunomodulator OM-8980 in rheumatoid arthritis was compared with that of auranofin, an oral gold salt, in a double blind, randomised multicentre study lasting six months. Seventy patients were treated with auranofin and 75 with OM-8980. The patients of both groups improved significantly at three and six months for all the clinical parameters observed: Ritchie index, number of swollen joints, morning stiffness, pain, grip strength, intake of non-steroidal anti-inflammatory drugs, and erythrocyte sedimentation rate. No serious side effects were observed in either group. The patients receiving auranofin had more adverse reactions, mainly affecting the gastrointestinal system. PMID- 3041925 TI - The development of new immunotherapies for the treatment of cancer using interleukin-2. A review. AB - Recent increases in knowledge of cellular immunology, combined with developments in biotechnology, have provided new opportunities for the development of immunotherapies for the treatment of cancer in humans. One approach to therapy is that of adoptive immunotherapy, that is, the transfer to the tumor bearing host of lymphoid cells with antitumor reactivity that can mediate antitumor responses. Several lymphocyte subpopulations have now been identified that may be suitable for use in adoptive immunotherapy. Resting lymphocytes incubated in interleukin-2 (IL-2) give rise to lymphokine activated killer (LAK) cells that can lyse malignant cells, but not normal cells. Clinical studies in patients with advanced cancer have revealed that treatment with high dose IL-2 alone or in combination with LAK cells can mediate the complete or partial regression of cancer in selected patients. Other approaches are currently undergoing investigation, including the adoptive transfer of tumor infiltrating lymphocytes, which, in animal models, have antitumor reactivity 50-100 times more potent than do LAK cells. Other new approaches to immunotherapy include the use of combination of lymphokines, such as the use of tumor necrosis factor or alpha interferon in conjunction with IL-2. The availability of recombinant lymphokines that provide large amounts of biologically active materials can hopefully lead to the development of effective new therapies for cancer in humans. PMID- 3041926 TI - Relationship between the diagnosis, preoperative evaluation, and prognosis after orthotopic liver transplantation. AB - The purpose of this study was to identify which of the biochemical, immunological, or functional parameters derived before surgery as part of a systemic evaluation were helpful in predicting the frequency of rejection episodes, the chance of survival, and the cause risk of death (should death occur) of patients after orthotopic liver transplantation (OLTx). Ninety-eight adult patients who had an extensive preoperative protocol evaluation were studied before OLTx. The biochemical parameters assessed were albumin, prothrombin time, bilirubin, and ICG clearance. The immunologic parameters assessed included total lymphocytes, T3 cells, T4 cells, T8 cells, and the T4/T8 ratio. The degree of histocompatibility antigen (HLA) matching between the donor and the recipient was also evaluated in 80 of the 98 patients studied. Most postoperative deaths occurred within 12 weeks of the procedure (24%; 24 of 98 patients); 13 patients (13%) died within the first 6 postoperative weeks, of either bacterial or fungal sepsis. An additional 14 patients (14%) died after the initial 6 postoperative weeks due, primarily of an acquired viral and/or protozoan infection (p less than 0.01). During the first 6 weeks, survival was better for patients with cholestatic liver disease (ChLD, 93%, n = 45) and miscellaneous liver diseases (MISC, 100%, n = 10) than it was for those with parenchymal liver diseases (PLD, 77%, n = 43). Although albumin, prothrombin time, T4/T8 ratios, and per cent T8 cells were statistically different in patients with PLD as compared with those with ChLD, these parameters, as well as the per cent T4 cells, serum bilirubin level, per cent retention of ICG at 15 minutes, and the plasma ICG disappearance rate were not found to be of substantial help in predicting patient survival or non-survival. Moreover, neither the degree of HLA matching nor the number of rejection episodes differed between surviving and nonsurviving patients. The results of this study suggest that patients with PLD are at increased risk of early postoperative death after OLTx because of bacterial and/or fungal sepsis, as compared with patients operated upon for ChLD. Better pre-, intra-, and postoperative predictors of risk of death and complications are needed to reduce the early mortality observed after orthotopic liver transplantation. PMID- 3041927 TI - A successful cardiac transplantation. PMID- 3041929 TI - Cutaneous leishmaniasis in Jordan: biochemical identification of human and Psammomys obesus isolates as Leishmania major. AB - As part of a series of epidemiological and ecological studies of leishmaniasis in Jordan, we have made functional studies of four isolates from human lesions and from ear lesions of three field-collected Psammomys obesus. Primary isolates were subcultured, frozen stabilates prepared and BALB/c mouse infectivity experiments initiated. Each mouse was inoculated with 4-8 x 10(4) promastigotes into a hind footpad. Quantitative evaluation of the footpads showed enlargement three to four weeks postinoculation. Amastigotes were readily identified in smears from footpad lesions and promastigotes in culture. At 47 days, liver and spleen samples grew out promastigotes. Biochemical characterization of these seven isolates was made by isozyme analysis using cellulose acetate membrane electrophoresis of fructokinase, phosphoglucose isomerase, phosphoglucomutase, aspartate aminotransferase, malate dehydrogenase, glucose-6-phosphate dehydrogenase and 6 phosphogluconate dehydrogenase. Reference isolates used for comparison were Leishmania major, L. tropica minor, L. donovani, L. aethiopica and L. m. mexicana. All seven Jordan isolates showed enzyme electromorphs identical to L. major, confirming our ecological/epidemiological studies that P. obesus is a major reservoir for human cutaneous leishmaniasis in Jordan. PMID- 3041928 TI - Decreased malaria morbidity in the Tharu people compared to sympatric populations in Nepal. AB - The Terai region of Nepal has been known to be heavily malarious since remote times, and it has, therefore, been regarded as uninhabitable by most Nepalese people. The Tharu people, who have been living in the Terai for centuries, were reputed to have an innate resistance to malaria. Following successful control of malaria by the Nepal Malaria Eradication Organization (NMEO), a large and heterogeneous non-Tharu population now inhabits the Terai along with Tharus. By analysing NMEO records, we have found that the prevalence of cases of residual malaria is nearly seven times lower among Tharus compared to sympatric non Tharus. This difference applies to Plasmodium vivax, which is now much more common, and to Plasmodium falciparum. We suggest that the basis for resistance to malaria in the Tharu people is a genetic factor yet to be identified. PMID- 3041930 TI - Primary cardiac echinococcosis: report of two cases with review of the literature. AB - Two case reports of Saudi patients with primary cardiac hydatid cysts are presented. In the first case, a multilocular cyst was located in the wall of the left ventricle, whereas in the second case a cyst was located in the pericardial sac and another cyst in the left ventricular wall. The diagnosis was based on a history of animal contact, full clinical examination, serological tests and the use of plain radiography, including conventional tomogram, two-dimensional echocardiography, computed tomography and thallium perfusion isotope scan. This study has indicated that non-invasive radiological methods are sufficient to diagnose cardiac echinococcosis and could provide the same information as, or even more than if invasive techniques were used. Further screening of the two patients showed no involvement of other organs by hydatid cysts. PMID- 3041931 TI - Filariasis in the Igwun River Basin, Nigeria: an epidemiological and clinical study with a note on the vectors. AB - In a cross-sectional, epidemiological and parasitological study of human filariasis, 845 individuals were examined in settlements along the Igwun Basin, Imo State, Nigeria. Four different filarial nematode species were identified. Two hundred and fifty-six (30.3%) of the individuals examined were positive for Onchocerca volvulus, 113 (13.4%) for Mansonella perstans, 76 (9.6%) for Wuchereria bancrofti and 77 (9.1%) for Loa loa. Microfilarial rates increased with age of individuals and showed a tendency towards higher prevalence rates in males than in females. The intensity of O. volvulus infection was high, with the highest microfilarial density of 44 mf mg-1 snip which occurred in the 40-49-year old individuals. In W. bancrofti and L. loa infections, infections of over 1000 mf 20 ml-1 blood were recorded in 15.8% and 19.5% of individuals, respectively. Observed clinical signs were associated with inflammatory, lympho-obstructive and ocular manifestations. In M. perstans infections all clinical cases were inflammatory. In W. bancrofti, 44.4% of clinical cases were inflammatory, and lympho-obstructive manifestations consisted of 23.8% chyluria, 12.7% hydrocele and 19.1% elephantiasis. In L. loa infections all clinical cases were inflammatory with indications of Calabar swellings. In O. volvulus infections 23.5% of clinical cases were inflammatory, while 76.5% showed ocular manifestations. The absence of blindness despite high O. volvulus infection rates was remarkable. The presence of potential insect vectors and the occurrence of clinical signs are indications of active transmissions. PMID- 3041932 TI - Transmission dynamics and estimates of malaria vectorial capacity for Anopheles balabacensis and An. flavirostris (Diptera: Culicidae) on Banggi island, Sabah, Malaysia. AB - Holoendemic malaria transmission in two small isolated forest communities and a coastal village was studied by (1) all night human bait collections of Anopheles species from inside and outside houses and (2) buffalo-biting and CDC light trapping catches during March and November 1984. During the same period thick and thin blood films were collected from the human population, and spleen rates were determined in children from two to nine years of age. Using both the immunoradiometric assay (IRMA) and the dissection technique, more sporozoite positive infections were detected in An. balabacensis and An. flavirostris in November than in March. IRMA confirmed the presence of Plasmodium falciparum sporozoites. An average of 76.2% of the An. balabacensis population lived long enough to have reached a point where infectivity with P. falciparum was possible in November. Although fewer than five adult females bit humans per night at any time, a resident could theoretically have received more than 160 infective bites in one year. A high frequency of feeding on humans, coupled with increased anopheline life expectancy, contributed to high estimates of falciparum malaria vectorial capacity (number of infections distributed per case per day); for An. balabacensis (1.44-7.44 in March and 9.97-19.7 in November) and for An. flavirostris (0.19-5.14 in March and 6.27-15.8 in November). These high values may explain the increased malaria parasite rates obtained from at least two forest communities. Correlation between actual and calculated rates of gametocytaemia was poorest in Kapitangan due to inadequate sampling of the human population. In Banggi island, malaria is stable and holoendemic, and the population enjoys a high degree of immunity. PMID- 3041933 TI - Bridge to cardiac transplantation. PMID- 3041934 TI - Bridging to cardiac transplantation with pulsatile ventricular assist devices. AB - As cardiac transplantation becomes more commonplace in the treatment of end-stage heart failure and as suitable donors become less available, an increasing number of patients will require mechanical circulatory assistance to bridge to transplantation. Since 1982, refractory hemodynamic instability requiring placement of pulsatile ventricular assist devices (VADs) has developed in 11 candidates for transplantation aged 24 to 54 years (mean, 39.6 years). A pneumatic Pierce-Donachy pump was used in 9 patients and an electrical Novacor pump in 2. The cause of the cardiomyopathy was ischemic in 6, postpartum in 2, idiopathic in 2, and doxorubicin hydrochloride toxicity in 1. Seven patients required left ventricular support (LVAD); 4 required biventricular mechanical support (BVAD). Duration of support ranged from 8 hours to 91 days with flows ranging from 4.1 to 8.5 L/min (mean, 5.5 L/min). Although hemodynamic stability was achieved in all 11 patients, contraindications to transplantation developed in 5 patients during VAD support (renal failure in 4, sepsis in 3, disseminated intravascular coagulopathy in 1). The remaining 6 patients (4 with an LVAD, 2 with a BVAD) remained good candidates for transplantation despite major complications in 5 (mediastinal bleeding in 3, driveline infection in 3, development of preformed antibodies in 2, small embolic stroke caused by device malfunction in 1). The 3 patients who were supported the longest (24, 75, and 91 days) were ambulatory while awaiting a donor heart. All 6 patients underwent successful transplantation after 8 hours to 91 days (mean, 24 days) of support. Other than one sternal wound infection, there were no major complications after transplantation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3041935 TI - Clinical evaluation of pressure-controlled intermittent coronary sinus occlusion: randomized trial during coronary artery surgery. AB - Pressure-controlled intermittent coronary sinus occlusion (PICSO) was evaluated in a randomized trial in 30 patients undergoing bypass surgery. PICSO was applied for one hour during early reperfusion. Myocardial function was determined from short-axis cross-sectional views of intraoperative two-dimensional echocardiography. Changes of sectional and segmental wall motion during extracorporeal circulation were analyzed. Although sectional wall motion did not change significantly, hypokinetic segments were preserved better in PICSO-treated patients than in controls (-1.3 +/- 2.4 versus -9.1 +/- 2.6 delta% fractional area change; p less than 0.04). Although not significant, the same trend was found for normal and severely hypokinetic segments. Cumulative enzyme release was related to coronary sinus occluded pressure (r = 0.94; p less than 0.006), indicating washout of metabolites during PICSO. Three months after operation, functional classification was similarly favorable in both groups. Long-term effects of PICSO cannot be predicted because PICSO was applied only during early reperfusion. We conclude that PICSO is a safe procedure and that its short-term beneficial effects on myocardial function suggest a preservation of myocardial viability. PMID- 3041936 TI - Remission of mild to moderate hypertension after treatment with carteolol, a beta adrenoceptor blocker with intrinsic sympathomimetic activity. AB - Previous studies have indicated that some hypertensive patients, following a period of effective treatment with certain antihypertensive drugs, may experience prolonged normotension after drug withdrawal. We have studied the ability of carteolol, a nonselective beta-adrenoceptor antagonist with intrinsic sympathomimetic activity, to produce such remissions of hypertension. Thirty-four patients whose diastolic blood pressure was controlled at 90 mm Hg or less with carteolol monotherapy (2.5 to 5.0 mg/d for an average of 328 days) were randomized to a nine-month, double-blind, placebo-controlled drug-withdrawal trial. Those patients randomized to continue carteolol therapy had initially responded to carteolol treatment with reduction in blood pressure from 151 +/- 4/99 +/- 2 to 132 +/- 4/80 +/- 2 mm Hg. Those randomized to treatment with placebo had initially responded with blood pressure reductions from 154 +/- 4/97 +/- 2 to 137 +/- 4/81 +/- 2 mm Hg. Changes in mean systolic and diastolic blood pressure (mm Hg +/- SEM) from baseline during carteolol therapy to the final visit at nine months were not different for patients receiving placebo (13 +/- 5/6 +/- 4 mm Hg, recumbent; 11 +/- 6/4 mm Hg, standing) or carteolol (11 +/- 5/7 +/- 3 mm Hg, recumbent; 12 +/- 6/7 +/- 3 mm Hg, standing). The final mean recumbent diastolic blood pressure (86.9 mm Hg) was the same in both groups. Prolonged normotension may follow a period of carteolol treatment, again suggesting the potential importance of periodic withdrawal of antihypertensive medication. PMID- 3041938 TI - Selective screening for abdominal aortic aneurysms with physical examination and ultrasound. AB - Abdominal aortic aneurysm (AAA) is an important cause of preventable death in older persons. Persistently high rupture mortality rates indicate that these deaths can be prevented only by early detection and treatment of AAA. In an effort to develop an effective and efficient program of AAA detection, we selectively screened a high-risk population. Men aged 60 to 75 years with hypertension and/or coronary artery disease were randomly selected from a general medicine clinic and screened with physical examination and ultrasound. Eighteen previously unsuspected aneurysms, 3.6 to 5.9 cm in size (mean, 4.4 cm), were detected in 201 patients, for a prevalence of 9% (95% confidence interval: 4.7% to 13.3%). The specificity and positive predictive value of ultrasound were each 100%. Abdominal palpation detected only half of these aneurysms, but missed none in patients with an abdominal girth less than 100 cm (n = 6). This degree of sensitivity did not occur with "routine" examinations and requires that the examination be directed specifically toward AAA detection. We conclude that undiagnosed AAAs are common in this large subgroup of the clinic population, that ultrasound is an excellent screening test for AAAs, and that physical examination may be adequate for screening thin patients. We recommend that every two or three years persons over the age of 50 years undergo careful abdominal palpation aimed at detecting AAAs, as part of the periodic health examination. We further recommend that obese older men at high risk for AAA have at least one-time screening with abdominal ultrasound, regardless of findings on physical examination. PMID- 3041937 TI - Improved metabolic control in insulin-dependent diabetes mellitus with insulin and tolazamide. AB - The influence of sulfonylurea drugs in enhancing the effect of endogenous insulin is well documented. Furthermore, combination therapy with sulfonylurea and insulin is effective in the treatment of type II diabetes mellitus. Therefore, to assess the efficacy of this type of combination therapy in type I diabetes, we conducted a double-blind clinical trial with tolazamide and insulin in 15 subjects with type I diabetes. The diagnosis of type I diabetes was confirmed by previous episodes of diabetic ketoacidosis and undetectable C-peptide levels in serum samples from blood drawn from patients two hours after breakfast. During the study protocol, placebo or tolazamide was randomly added to insulin and the combination therapy was continued for three months. In the placebo group, levels of fasting plasma glucose (FPG) and hemoglobin A1c (HbA1c) did not alter significantly at the end of the study period. However, in the tolazamide group, levels of FPG and HbA1c markedly improved after administration of tolazamide (FPG levels before therapy, 10.8 +/- 0.9 mmol/L [mean +/- SEM]; after therapy, 6.7 +/- 0.4 mmol/L; HbA1c levels before therapy, 10.9% +/- 0.6%; after therapy, 9.6% +/- 0.5%). Therefore, adjuvant therapy with tolazamide and insulin may be beneficial in achieving adequate metabolic control in type I diabetes mellitus. PMID- 3041939 TI - Noninvasive evaluation of the extracranial carotid arteries in patients with cerebrovascular events and atrial fibrillations. AB - Noninvasive carotid artery testing was performed in 73 patients with nonvalvular atrial fibrillation who were referred because of symptoms or signs of cerebrovascular disease. Thromboembolism related to atrial fibrillation without valvular heart disease was the probable source of cerebral ischemia in 25 (80%) of 31 patients with stroke and coexisting atherosclerotic disease at the carotid artery bifurcation in six (20%). Nonvalvular atrial fibrillation was the probable source of symptoms in nine (70%) of 13 of patients with transient cerebral ischemia, while coexisting carotid artery disease was present in four (30%). Nonvalvular atrial fibrillation accounted for the symptoms in four of five patients with amaurosis fugax, with atherosclerotic carotid artery disease present in one. The remaining 24 patients had nonhemispheric symptoms of cerebrovascular disease, including vertebrobasilar insufficiency, dizziness, and syncope, and only one had a carotid lesion. A significantly higher proportion of patients with focal hemispheric symptoms had coexisting carotid disease than patients with nonfocal symptoms had, suggesting that atherosclerotic cerebrovascular disease contributes to stroke in patients with nonvalvular atrial fibrillation. Noninvasive carotid artery testing may be helpful in identifying atherosclerotic lesions at the carotid artery bifurcation in patients with atrial fibrillation and cerebrovascular disease, because different therapeutic modalities may be appropriate when two potential sources of cerebral ischemia are present. PMID- 3041940 TI - The effect of intravenous magnesium therapy on serum and urine levels of potassium, calcium, and sodium in patients with ischemic heart disease, with and without acute myocardial infarction. AB - Serum concentrations of magnesium, potassium, calcium, and sodium were determined on admission of 224 patients to the hospital and after 2, 4, and 6 days in hospital; all were admitted to the hospital with suspected acute myocardial infarction (AMI). On admission, the patients were randomly allocated to 48 hours of treatment with magnesium intravenously or placebo. One hundred twenty-three patients had AMI (of whom 53 [43%] were treated with magnesium) and 101 had their suspected AMI disproven (of whom 51 [50%] were treated with magnesium). In a supplementary study, serum and urine levels of magnesium, potassium, calcium, and sodium, together with serum levels of parathyroid hormone, were determined before and after intravenous magnesium treatment in six patients with AMI and six patients with ischemic heart disease but without AMI. In both studies, magnesium therapy was associated with significant alterations in extracellular ion homeostasis. Serum concentrations of potassium decreased during the initial days of hospitalization in the patients treated with placebo, but increased slightly in the patients treated with magnesium infusions. These increments in the serum concentrations of magnesium and potassium correlated significantly. The increase in the serum concentration of potassium after magnesium infusions was due to a reduced renal potassium excretion level (from 71.3 to 49.4 mmol/24 h), indicating the existence of a divalent-monovalent cation exchange mechanism in the nephron. This hypothesis was supported by the observation that renal sodium excretion likewise decreased after magnesium infusions (from 83.2 to 59.2 mmol/24 h). Serum concentration of calcium decreased significantly after magnesium treatment (from 2.35 mmol/L on admission to 2.15 mmol/L after 24 hours in the hospital) in the AMI group, in contrast to the placebo-treated patients, where no significant fluctuations in serum concentration of calcium were detected during the initial six days. This decrease in serum concentration of calcium was due to a marked increase in renal calcium excretion (from 3.43 mmol/24 h before to 6.59 mmol/24 h after magnesium infusion). A correlation between increments in serum magnesium concentration and decrements in serum calcium concentration was detected. No change in serum levels of parathyroid hormone was found before and after magnesium infusions. Both serum and urine levels of magnesium significantly increased after magnesium treatment to levels above the upper normal limits (serum magnesium concentration increased from 0.81 to 1.21 mmol/L, urine magnesium excretion levels from 3.57 to 16.57 mmol/24 h for both serum and urine changes.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3041942 TI - The Optic Neuritis Treatment Trial. PMID- 3041941 TI - Use of animals in biomedical research. Historical role of the American Medical Association and the American physician. AB - From the introduction of the "Gallinger-DC" bill in 1896 to the passage of the Laboratory Animal Welfare Act in 1966, organized medicine and the American physician have been active in promoting the humane and appropriate use of research animals and explaining to the public and legislators the importance of research using animals to medical progress. The role of organized medicine and science in events leading to passage of federal legislation is discussed. Past efforts of the American Medical Association and the American physician have been critical in numerous successful efforts at the local, state, and national level to prevent the passage of laws which restricted animal use for health research and impeded medical progress. This article demonstrates that current initiatives by physicians to preserve biomedical research are a reaffirmation of their traditional role. PMID- 3041943 TI - Cutaneous malignant melanoma in survivors of heritable retinoblastoma. AB - Six survivors of bilateral retinoblastoma developed cutaneous malignant melanoma 20 to 51 years (average, 31 years) after initial therapy for the ocular tumor. Five patients received radiation therapy to the orbital area. In two patients the cutaneous malignant melanoma developed in the field of irradiation. Two patients developed multiple cutaneous melanomas and are thought to have the dysplastic nevus syndrome. At this writing two patients are alive and well after wide resection of their skin tumors. A review of the literature suggests that cutaneous malignant melanoma accounts for about 7% of second malignant neoplasms in survivors of heritable retinoblastoma. PMID- 3041944 TI - Apraclonidine. A one-week dose-response study. AB - We performed a double-masked, cross-over, dose-response study of apraclonidine hydrochloride (formerly known as ALO 2145) in 20 patients with elevated intraocular pressure (IOP). We administered three concentrations of apraclonidine (0.125%, 0.25%, 0.5%) and vehicle alone bilaterally every 12 hours for one week. Patients were examined 2, 5, and 8 hours after the initial dose, and then on day 2 and day 8. We studied IOP, pupillary diameter, interpalpebral fissure width, blood pressure, and pulse. There was a two-week washout period after each one week session. All concentrations of apraclonidine significantly lowered IOP. The 0.5% and 0.25% concentrations had equal maximal effects, lowering IOP in each patient by an average of 27% relative to vehicle alone. This corresponded to a mean decrease in IOP of 8.7 mm Hg, from a baseline of 24.9 mm Hg to 16.2 mm Hg. The 0.5% and 0.25% concentrations were significantly more effective than the 0.125% concentration at two and eight hours. Mean interpalpebral fissure width increased in a dose-dependent fashion; the pupillary effect was minimal. Blood pressure and pulse were unchanged. Thirty percent of subjects reported transient dry nose or dry mouth. These symptoms may be dose-dependent. PMID- 3041945 TI - Vasoactive intestinal polypeptide and the innervation of the human lacrimal gland. AB - Vasoactive intestinal polypeptide (VIP) is a biologically active neuropeptide found in both the peripheral and the central nervous systems. Previous studies have shown that VIP-like immunoreactive nerves are present in the uveal tissues of the human eye. The distribution of VIP-like immunoreactivity of the human lacrimal gland and sphenopalatine ganglion was studied. A lacy network of VIP like immunoreactive nerve fibers was found in the lacrimal gland and was predominantly located along the basilar surface of the acinar epithelium and in the interstitial connective tissue of the gland. This pattern of innervation was nearly identical to the distribution of cholinesterase-positive fibers in human lacrimal glands. The VIP-like immunoreactive cell bodies were found throughout the sphenopalatine ganglion obtained at autopsy. The distribution of VIP-like immunoreactive nerves in the human lacrimal gland and sphenopalatine ganglion was generally similar to that described in mammalian and avian systems, although some differences were noted. Vasoactive intestinal polypeptide may represent an important cotransmitter or neuromodulator for the facial parasympathetic nerves that supply the eye and the lacrimal gland. PMID- 3041946 TI - [Abnormalities of the petrous bone and the adjacent base of the skull]. PMID- 3041947 TI - [Surgery of the petrous bone (middle ear). Tumors and pseudotumors]. PMID- 3041948 TI - [Tumors and pseudotumors of the petrous bone and adjacent base of the skull. Neurosurgical reference]. PMID- 3041949 TI - [Threatening inflammations of the petrous bone and adjacent base of the skull]. PMID- 3041950 TI - [Injuries and fractures of the petrous bone and adjacent base of the skull]. PMID- 3041951 TI - A Clostridium botulinum type B vaccine for prevention of shaker foal syndrome. AB - A toxoid was prepared from type B toxin of Clostridium botulinum by treatment with 0.6% formalin for 6 weeks. The toxoid was adsorbed to aluminium hydroxide and this vaccine was evaluated for safety in guinea pigs, mice and horses, and for immunogenicity in guinea pigs and horses. Neutralising antitoxin was demonstrated in adult horses receiving two 2 ml subcutaneous doses 6 weeks apart, and in a foal which suckled its vaccinated dam. Another vaccinated mare and the passively immunised foal were protected against subcutaneous injection of 1600 and 2000 mouse lethal doses of toxin per kg respectively. PMID- 3041952 TI - University of Queensland School of Veterinary Science--looking both ways. PMID- 3041953 TI - Comparison of direct electron microscopy and enzyme immunoassay for the detection of rotaviruses in calves, lambs, piglets and foals. AB - Direct electron microscopy (EM) and enzyme-immunoassay (rotazyme) results for the detection of rotaviruses in 346 enteric specimens from calves, lambs, piglets and foals were compared. The rotazyme test was at least 3 times more sensitive than direct EM in diagnosing infection. Rotavirus antigen was demonstrated by rotazyme in 22% of 280 scour samples and in 27% of 66 samples from non-scouring animals. There was an association between diarrhoea and higher amounts of rotavirus antigen. This prevalence of rotaviruses detected in animals with diarrhoea highlights the significant involvement of other pathogens identified in the study including Eimeria, Cryptosporidia, Escherichia coli, Campylobacter, and other viruses. PMID- 3041954 TI - A Johne's disease survey and comparison of diagnostic tests. PMID- 3041955 TI - The influence of physical and environmental variables on the in vitro attachment of Salmonella typhimurium to the ceca of chickens. AB - This investigation was designed to study the effect of exposure time, pH, age of bird, and native intestinal microflora on the in vitro attachment of Salmonella typhimurium to the ceca of chickens. Ceca were surgically removed from chickens immediately after euthanasia, and the interiors were exposed to S. typhimurium as intact ceca with contents, intact ceca rinsed free of contents, or inverted rinsed ceca. Attachment of S. typhimurium was slightly higher in washed than in unwashed ceca. Neither pH nor age of chicks affected attachment of the organism to ceca. There was no difference in attachment of salmonellae to inverted washed ceca after 1, 2, 3, 5, and 10 min exposure, but a one log difference was noted between 10 min and 30 sec. PMID- 3041956 TI - Lesions of right heart failure and ascites in broiler chickens. AB - A naturally occurring cardiomyopathy in broiler chickens from a single Ontario farm was studied in order to define the morphologic changes. Gross and histologic features of affected birds were compared with those in age-matched control penmates. Body weight and weight and volume of individual cardiac chambers were measured. Histologic sections of 18 different tissues were examined, and lesions observed were scored subjectively. Affected birds were stunted and had marked right ventricular dilation and hypertrophy, atrial hypertrophy, ascites, pulmonary congestion and edema, and hepatic capsular fibrosis. Microscopic changes in the heart of affected birds were mild and did not suggest a specific cause of heart failure. Lungs had marked hypertrophy of parabronchial smooth muscle and collapse and apparent loss of associated air capillaries. Other histologic changes observed were thought to be the result of passive congestion of viscera caused by right heart failure and chronic debility. Although the specific etiology of this condition could not be determined, it was felt that this syndrome was unlikely to have been the result of any of the commonly recognized causes of congestive heart failure and ascites in broilers. PMID- 3041957 TI - Evaluation of the specificity and sensitivity of two commercial enzyme-linked immunosorbent assay kits, the serum plate agglutination test, and the hemagglutination-inhibition test for antibodies formed in response to Mycoplasma gallisepticum. AB - Two commercial enzyme-linked immunosorbent assay (ELISA) kits, seven serum plate agglutination (SPA) antigens, and the hemagglutination-inhibition (HI) test for antibodies to Mycoplasma gallisepticum (MG) were compared for sensitivity and specificity using known MG-positive and MG-negative sera from leghorn chickens. All SPA antigens proved to be highly sensitive when testing MG-positive sera. Laboratory-prepared SPA antigens yielded fewer positive reactions when testing MG negative sera than commercial SPA antigens. Both MG ELISA kits showed high rates of positive reactions when testing sera from birds given commercial M. synoviae bacterin, fowl coryza (Haemophilus paragallinarum) bacterin, inactivated infectious bursal disease virus vaccine, and to a lesser extent fowl cholera (Pasteurella multocida) bacterin. Immunization with Frey's medium with 12% swine serum-in-oil or Staphylococcus aureus-in-oil resulted in sera which yielded numerous positive ELISA reactions. During the first 1 to 3 weeks, antibodies induced by experimental infection with MG were better detected by the SPA test than by the ELISAs and the HI test, thus confirming the SPA test's importance in Mycoplasma diagnostic serology. The HI test can serve to confirm positive SPA results. PMID- 3041959 TI - DSM-III-R: too much too soon? AB - Some of the more important changes carried out in the recently published revision of the DSM-III classification (DSM-III-R) are described, and an assessment is made on whether a revision of this nature was necessary and whether it achieved its aims. Since a large number of disorders have been modified in one way or another and since empirical evidence for many of the changes is lacking, the author concludes that DSM-III-R, rather than illuminating the field, is likely to cause confusion and frustration in the minds of clinicians, researchers and administrators, and that in fact DSM-III-R is a new classification, a DSM-IV in disguise. PMID- 3041958 TI - Effect of anticoccidial and antimicrobial feed additives on prevention of Salmonella colonization of chicks treated with anaerobic cultures of chicken feces. AB - Chicks were treated orally on the day of hatch with either fresh or frozen competitive exclusion (CE) cultures (native gut microflora). Chicks were fed either unmedicated feed or one of five commercial broiler starter rations or nine experimental feed mixtures containing varying amounts and combinations of anticoccidial and antimicrobial medicaments. After 2 days, they were challenged with approximately 10(6) colony-forming units of a nalidixic-acid-resistant strain of Salmonella typhimurium. Six days later, chicks were sacrificed and ceca were analyzed for S. typhimurium. Colonization of 2-day-old chicks was prevented or at least greatly reduced in most instances by treatment of chicks with a CE culture, but the efficacy of CE broth cultures stored at -70 C diminished over time. Not all CE cultures tested gave equal protection against Salmonella colonization, and CE cultures were more susceptible to some feed additives than others. Of the commercial or experimental feed tested, only the feed containing the combination of nicarbazin and bacitracin interfered with the protective effect of the CE culture. PMID- 3041960 TI - Psychosocial causes of duodenal ulcer. AB - The evidence that duodenal ulcer is a psychosomatic disorder is reviewed. A variety of both experimental stressors and differing states of emotional arousal (anxiety and anger in particular) are associated with increased acid and pepsin secretion. Furthermore, chronic life stressors predispose to duodenal ulcer, presumably by way of symptomatic anxiety or depression, while poor social support may comprise another risk factor. Subjects with high levels of trait anxiety or neuroticism are more prone to stress-induced anxiety and its physiological consequences. There is minimal acceptable evidence to support more specific personality-related theories, including Type A behaviour, alexithymia or the earlier psycho-analytic theories of Alexander. Nonetheless, it is concluded that Alexander's inclusion of duodenal ulcer as one of a handful of classic psychosomatic disorders has been confirmed. Since some 40% of duodenal ulcer patients relapse in the first year after medical treatment, there may be some role for psychological treatments (especially in the domain of stress/anxiety management) for those patients who relapse and have co-existing high state or trait anxiety or exposure to chronic stressors. PMID- 3041961 TI - Proteoglycans of the human intervertebral disc. Electrophoretic heterogeneity of the aggregating proteoglycans of the nucleus pulposus. AB - Nuclei pulposi were dissected from lumbar discs of radiologically normal human spines of cadavers aged 17, 20 and 21 years. Proteoglycans were extracted with 4 M guanidine hydrochloride (dissociative conditions) with proteinase inhibitors and isolated as A1 fractions by associative density-gradient centrifugation. Aggregating and non-aggregating proteoglycans were separated by Sepharose 2B chromatography. Both aggregating and non-aggregating proteoglycans contained a keratan sulphate-rich region as isolated by chondroitinase/trypsin/chymotrypsin digestion and Sepharose CL-6B chromatography. Agarose/acrylamide-gel electrophoresis of individual fractions of a Bio-Gel A-50m dissociative-column separation of the aggregating proteoglycans revealed two, well-separated bands: S and F, the slower and faster migrating bands respectively. The non-aggregating proteoglycan fractions were eluted under associative conditions (0.5 M-sodium acetate, pH 6.8) and migrated as a single band in the electrophoretic system. The gel-electrophoretic heterogeneity of the aggregating proteoglycans was still evident after hydroxylamine fragmentation and removal of the hyaluronate-binding portion of the molecule. Dissociative density-gradient centrifugation of the aggregating proteoglycans partially separated the Band-S proteoglycans from the Band-F population. Subsequent dissociative chromatography of the high-buoyant density Band F proteoglycans permitted discrimination of this band into two gel electrophoresis-distinguishable populations (Bands F-1 and F-2). Enzyme-linked immunosorbent assays with a monoclonal antibody that recognized keratan sulphate demonstrated that the D1 fraction containing the Band F-1 proteoglycans was enriched in keratan sulphate compared with the total aggregating or non aggregating pool of proteoglycans. The proteoglycans of young adult nucleus pulposus could then be ascribed to one of four structurally and/or electrophoretically distinct populations: (1) the non-aggregating population, which comprised about 70% of the total extractable proteoglycans; (2) the aggregating pool, comprising: (a) Band F-1 proteoglycans, which had a relatively large hydrodynamic size, uronate/protein weight ratio, were enriched in keratan sulphate and had a high buoyant density; (b) Band S proteoglycans, which migrated slower in agarose/acrylamide gels, had a smaller hydrodynamic size, lower buoyant density and a lower uronate/protein ratio than the Band F-1 population; (c) Band F-2 proteoglycans, which were lower in buoyant density, smaller in hydrodynamic size and slightly faster in electrophoretic mobility than the Band F-1 proteoglycans. PMID- 3041962 TI - Regulation of inositol 1,4,5-trisphosphate metabolism in insulin-secreting RINm5F cells. AB - Factors underlying the transience of inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] accumulation following muscarinic stimulation of RINm5F cells were examined. Transience was not due to a protein kinase C-mediated stimulation of Ins(1,4,5)P3 dephosphorylation, since pretreatment of cells with tetradecanoyl-phorbol acetate (TPA) did not alter the rate of this conversion. However, preincubation with TPA did inhibit carbamoylcholine-stimulated Ins(1,4,5)P3 formation. In permeabilized cells, the conversion of Ins(1,4,5)P3 to inositol 1,3,4,5-tetrakisphosphate [Ins(1,3,4,5)P4] was slightly enhanced in the presence of TPA or cyclic AMP, but much more markedly by raising the Ca2+ concentration from 10(-7) M to 10(-6) or 10(-5) M. In intact cells the most rapid rate of accumulation of Ins(1,4,5)P3 and Ins(1,3,4,5)P4 occurred in the first 2 s following stimulation, whereas the levels of inositol 1,4-bisphosphate were not increased until after 5 s. This suggests that Ins(1,4,5)P3 kinase is chiefly responsible for the early disposal of Ins(1,4,5)P3 following cellular stimulation. The results are consistent with the proposal that the transient accumulation of Ins(1,4,5)P3 is due both to its enhanced metabolism via the Ca2+-calmodulin-sensitive Ins(1,4,5)P3 kinase, as well as a down-regulation of phosphatidylinositol 4,5-bisphosphate hydrolysis. PMID- 3041963 TI - Purification and characterization of diacetyl-reducing enzymes from Staphylococcus aureus. AB - A method was developed to purify diacetyl-reducing enzymes from Staphylococcus aureus. Two enzymes capable of catalysing diacetyl reduction were isolated, neither of which turned out to be a specific diacetyl reductase. One of them is a lactate dehydrogenase similar to the one from Staphylococcus epidermidis, which accepts diacetyl, although poorly. The other one uses as coenzyme beta-NAD and reduces uncharged alpha-dicarbonyls with more than three carbon atoms (especially the alpha-diketones diacetyl and pentane-2,3-dione), producing the L(+) form of the corresponding alpha-hydroxycarbonyls. This enzyme has an Mr of 68,000 and is, most probably, a monomer. Its optimum pH is 6.0. Its shows a high affinity for NADH and a rather low one for diacetyl, which, at least in vitro, does not seem to be as good a substrate as pentane-2,3-dione. We propose for it the systematic name L-alpha-hydroxyketone:NAD+ oxidoreductase and the recommended name of alpha diketone reductase (NAD). We also suggest that the diacetyl reductase entry in the I.U.B. classification be suppressed. PMID- 3041965 TI - A tissue-specific increase in lipogenesis in rat brown adipose tissue in hypothyroidism. AB - 1. Rats were made hypothyroid by feeding them with propylthiouracil together with a low-iodine diet for 4 weeks. 2. [U-14C]Glucose conversion into fatty acids was substantially enhanced in brown adipocytes isolated from hypothyroid rats. Incorporation of 3H2O into fatty acids in vivo was enhanced in hypothyroidism in interscapular brown fat, but not in epididymal white fat or in liver. Hypothyroidism increased the activities of fatty acid synthase and ATP citrate lyase in brown, but not in white, adipocytes. 3. Glycolytic flux in brown adipocytes, quantified by [3-3H]glucose detritiation, was increased by hypothyroidism. This change was accompanied by increased maximum activity of phosphofructokinase. In white adipocytes a large increase in phosphofructokinase maximum activity was observed in hypothyroidism, but this change was accompanied by only small increases in the rate of glucose detritiation by incubated cells. It is suggested that in the brown adipocyte the overall conversion of glucose into fatty acids is enhanced in thyroid deficiency, but that this change is muted in the white adipocyte, possibly because of limitation of glucose transport. 4. Fatty acid synthesis in brown adipocytes from hypothyroid animals was considerably less sensitive to inhibition by exogenous fatty acids than is the process in cells from euthyroid animals. Consequently, the effect of hypothyroidism to enhance lipogenesis is amplified in the presence of physiological concentrations of fatty acid. PMID- 3041964 TI - Cycloheximide decreases glucose transporters in rat adipocyte plasma membranes without affecting insulin-stimulated glucose transport. AB - This study examines the relationship between insulin-stimulated glucose transport and insulin-induced translocation of glucose transporters in isolated rat adipocytes. Adipose cells were incubated with or without cycloheximide, a potent inhibitor of protein synthesis, for 60 min and then for an additional 30 min with or without insulin. After the incubation we measured 3-O-methylglucose transport in the adipose cells, and subcellular membrane fractions were prepared. The numbers of glucose transporters in the various membrane fractions were determined by the cytochalasin B binding assay. Basal and insulin-stimulated 3-O methylglucose uptakes were not affected by cycloheximide. Furthermore, cycloheximide affected neither Vmax. nor Km of insulin-stimulated 3-O methylglucose transport. In contrast, the number of glucose transporters in plasma membranes derived from cells preincubated with cycloheximide and insulin was markedly decreased compared with those from cells incubated with insulin alone (10.5 +/- 0.8 and 22.2 +/- 1.8 pmol/mg of protein respectively; P less than 0.005). The number of glucose transporters in cells incubated with cycloheximide alone was not significantly different compared with control cells. SDS/polyacrylamide-gel-electrophoretic analysis of [3H]cytochalasin-B photolabelled plasma-membrane fractions revealed that cycloheximide decreases the amount of labelled glucose transporters in insulin-stimulated membranes. However, the apparent molecular mass of the protein was not changed by cycloheximide treatment. The effect of cycloheximide on the two-dimensional electrophoretic profile of the glucose transporter in insulin-stimulated low-density microsomal membranes revealed a decrease in the pI-6.4 glucose-transporter isoform, whereas the insulin-translocatable isoform (pI 5.6) was decreased. Thus the observed discrepancy between insulin-stimulated glucose transport and insulin-induced translocation of glucose transporters strongly suggests that a still unknown protein-synthesis-dependent mechanism is involved in insulin activation of glucose transport. PMID- 3041968 TI - Bone marrow therapy with monoclonal antibodies. PMID- 3041966 TI - A protein-sialyl polymer complex involved in colominic acid biosynthesis. Effect of tunicamycin. AB - A protein-NeuAc complex involved in colominic acid biosynthesis has been identified in membrane preparations of Escherichia coli K-235. This compound had an Mr (estimated by SDS/polyacrylamide-gel electrophoresis and autoradiography) of about 100,000 and played the role of an 'initiator' or 'primer' (endogenous acceptor) in the synthesis of the whole polymer. Incubations of E. coli membranes with CMP-[14C]NeuAc (CMP-N-[14C]acetylneuraminic acid) pointed to the existence of a protein fraction (primer acceptor) that linked residues of sialic acid (N acetylneuraminic acid, NeuAc) up to a maximal size, later releasing them as low Mr sialyl polymers (LMrS, Mr less than 10,000). In the presence of colominic acid (final acceptor) the radioactivity linked to the protein quickly decreased, appearing stoichiometrically bound to the whole polysaccharide. When membrane preparations were previously digested with Streptomyces proteinase or de activated by heating (80 degrees C, 10 min), no incorporation of labelled NeuAc into trichloroacetic acid-insoluble material was detected. These results suggested that colominic acid molecules are synthesized while they are bound to a proteinaceous acceptor that is subsequently excised in the presence of colominic acid, generating the native protein. The antibiotic tunicamycin inhibited the biosynthesis of colominic acid, affecting the synthesis of this protein-(NeuAc)n intermediate. All these results are described here for the first time. PMID- 3041967 TI - Hormonal regulation of rat foetal lipogenesis in brown-adipocyte primary cultures. AB - Insulin stimulates lipogenesis by 100% for 5 h by a covalent modulation of acetyl CoA carboxylase, and by 200% for 24 h by increasing malic enzyme and fatty acid synthase enzymic activities in brown-adipocyte primary cultures. At short times, noradrenaline and isoprenaline decrease lipogenesis. However, phenylephrine and glucagon have no effect. At long times, dexamethasone inhibits lipogenesis. This effect is precluded in the presence of insulin. Progesterone and tri iodothyronine, alone or in the presence of insulin, produce a stimulation of the rates of lipogenesis. PMID- 3041969 TI - Effect of nutritional imbalances on cytochrome P-450 isozymes in rat liver. AB - Male Sprague-Dawley rats were fed for six weeks either a control diet containing 22% casein (C) and 5% fat (F) or a low-protein diet (6% C, 5% F) or high-lipid diet (30% C, 30% F). A group of rats received a control diet containing 50 ppm of Phenoclor DP6. Three major forms of cytochrome P-450, UT 50, BP 3a and MC 2 were purified from livers of DP6-fed rats and only two forms, UT 50 and PB 3a, were purified from control and dietary groups. The amino acid composition and the catalytic activities towards all substrates tested were only significantly modified in the purified UT 50 P-450 isozyme from rats fed the low-protein diet. The N-terminal sequence analysis shows that cytochrome P-450 UT 50 (from control group) and UT 501 (from low-protein group) are two distinct proteins. PMID- 3041970 TI - Long-lasting hypotensive and antihypertensive effects of a new 1,5 benzothiazepine calcium antagonist in hypertensive rats and renal hypertensive dogs. AB - The hypotensive effect of a new 1,5-benzothiazepine derivative, TA-3090 ((+) (2S,3S)-3-acetoxy-8-chloro-5-(2-(dimethylamino)ethyl)-2, 3-dihydro-2-(4 methoxyphenyl)-1,5-benzothiazepin-4-(5H)-one maleate), was studied in various models of experimental hypertension. In conscious spontaneously hypertensive rats (SHR), TA-3090 at a dose of 3 mg/kg p.o. or more caused significant hypotension. The effect was long-lasting, being observed for more than 8 h and 24 h at doses of 30 and 100 mg/kg p.o., respectively. Heart rate was not increased with doses up to 30 mg/kg p.o. TA-3090 like diltiazem, showed more potent hypotensive effect in SHR than in Wistar Kyoto rats (WKY). The enhancement of the hypotensive effect in SHR was more evident with TA-3090 and diltiazem than with 1,4-dihydropyridine calcium antagonists (1,4-DHPs). The enhanced hypotensive effect of TA-3090 was also observed in deoxycorticosterone acetate (DOCA)-salt hypertensive rats in which the minimum hypotensive dose of TA-3090 was 1 mg/kg p.o. In SHR, reflex tachycardia induced by TA-3090 was smaller than those induced by 1,4-DHPs, while diltiazem decreased the heart rate. In WKY, all calcium antagonists except diltiazem at a low dose (30 mg/kg p.o.) increased the heart rate. Development of hypertension in young SHR was significantly suppressed by chronic treatment with TA-3090 at a daily dose of 10 mg/kg p.o. or more. The antihypertensive effect of TA-3090 was also observed by administration in the diet in matured SHR. In addition, TA-3090 exhibited a dose-related natriuretic but not kaliuretic effect in SHR at doses slightly higher than the hypotensive ones.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3041971 TI - Immunopharmacological studies of new 3-benzoyl-4-mercaptobutyric acids. Immunomodulating effects. AB - A number of D-penicillamine (PA) derivatives (3-benzoyl-4-mercaptobutyric acids) having acetylthio groups on an alpha or beta position of a carboxylic acid, were synthesized and examined for their immunological effects compared with PA. New PA derivatives suppressed adjuvant-induced arthritis (AA) in SD rats and enhanced AA in Lewis rats like PA. Suppressive effects of 2-acetylthiomethyl-3-(4-methyl benzoyl)propionic acid (compound II-3) on AA in SD rats was most potent among PA derivatives and PA. II-3 enhanced type II collagen-induced arthritis in rats more effectively than PA, and it slightly prolonged the survival time of NZBXNZW hybrid (BWF1) mice. Hemolytic plaque forming cells in the spleen cells of BDF1 and aged Balb/c mice were potentiated but those of BWF1 were suppressed by both compounds. In in vitro experiments, both compounds enhanced lymphocyte transformation. On the contrary, II-3 had no effect on the acute inflammatory response, delayed type hypersensitivity and IgE antibody response. The abnormal release of lysosomal enzymes from the peritoneal macrophages of aged MRL/l mice were suppressed by both compounds. These results suggest that II-3 is an immunomodulator like PA but more effective than PA. II-3 may be clinically effective for rheumatoid arthritis. PMID- 3041973 TI - [Therapy of common cold with a homeopathic combination preparation in comparison with acetylsalicylic acid. A controlled, randomized double-blind study]. AB - A clinical test was carried out on 170 West German army soldiers suffering from common cold. The test was conducted on a monocentric, randomized, non-sequential, and inter-individual basis; the research workers were kept blind on the identity of the medication. The purpose of testing was to compare the effectiveness of a combination homeopathic preparation (Gripp-Heel) with that of acetylsalicylic acid. On the 4th and 10th treatment days, no significant difference was determined with respect to changes in clinical findings, subjectively assessed complaints, or length of time the patients were unable to work. Thus the two preparations possess comparative effectiveness in the treatment of the common cold. PMID- 3041974 TI - The value of women's work. PMID- 3041972 TI - Comparison of midazolam and oxazepam as hypnotics in elderly hospitalized patients. A double-blind clinical trial. AB - The efficacy and safety of 15 mg midazolam versus 15 mg oxazepam were compared in a double-blind clinical trial in 61 aged hospitalized patients (mean age 82.5, range 69 to 96), suffering from moderate to severe insomnia. The study covered 8 nights, i.e., 1 placebo night followed by 5 treatment nights and 2 further placebo nights. Both drugs were equally effective with respect to total sleep time, number of nocturnal awakenings, quality of sleep, condition on awakening, patients' assessment and dreams. However, sleep latency was significantly shorter in the midazolam group, which was also rated significantly more favorably by the medical staff. Side effects were mild and similar in both groups. No rebound or carry-over effects were noted. Midazolam can be considered an effective and safe sleep inducer in aged patients. PMID- 3041975 TI - ASHA women: education and jobs. PMID- 3041976 TI - Where have all the young men gone? PMID- 3041977 TI - Women's issues: an annotated bibliography. PMID- 3041978 TI - Cochlear neurobiology: revolutionary developments. PMID- 3041979 TI - Time management in clinical supervision. A descriptive study of time allocation. PMID- 3041981 TI - Serum cholesterol variations in individual patients. AB - Intra-individual variations of serum cholesterol levels were examined retrospectively over a period of 1-12 years in 77 office patients. The findings confirm for patients the variations published in the literature on volunteers. There was apparently more than one population, one with small variances and another with much greater variances. Because of the large variances in many patients, a physician should obtain several independent measures before attempting to evaluate any therapeutic program. PMID- 3041980 TI - Cell cycle-dependent inhibition of DNA synthesis by prostaglandin I2 in cultured rabbit aortic smooth muscle cells. AB - The role of prostaglandin I2 (PGI2) in the control of DNA synthesis during the cell cycle was investigated in cultured rabbit aortic smooth muscle cells (SMC). SMC at confluency in the G0 state reached the S phase about 16 h after stimulation with serum, as judged by measurement of [3H]thymidine incorporation into DNA (DNA synthesis). Cyclooxygenase inhibitors such as indomethacin and aspirin enhanced DNA synthesis, suggesting that endogenously synthesized prostaglandins inhibit DNA synthesis. Added PGE1 or PGE2 had little effect on DNA synthesis. PGI2 inhibited DNA synthesis only when added from 10 to 16 h after stimulation of SMC in the G0 state with serum. Addition of CS-570, a stable PGI2 analogue, inhibited DNA synthesis at any time after serum stimulation. The endogenous syntheses of PGI2 and DNA were negatively correlated. These results suggest that PGI2 inhibits DNA synthesis by acting on the progression stage of the G1 state. PMID- 3041982 TI - Extracoronary atherosclerosis in familial hypercholesterolemia. AB - Sixty-two patients (31 males, 31 females) with familial hypercholesterolemia (FH) underwent a vascular examination by Doppler ultrasound. The ankle/arm systolic pressure index was determined, and serum lipoproteins were analyzed. Eight of 124 legs examined (6.5%) showed an ankle/arm pressure index less than 0.95, suggesting flow reducing stenosis. Common carotid, internal carotid, and iliac arteries were evaluated by echo Doppler examination with spectral analysis. Forty five of the 372 arteries examined (12.1%) had detectable abnormalities at echo Doppler examination. Iliac and internal carotid artery lesions were significantly (P less than 0.01) more frequent among FH patients than in a control group (30 men, 20 women) comparable for sex and age. The mean age of the patients with heterozygous FH and detectable arterial lesions was 45.3 years and that of those without lesions 30.7 years (P less than 0.05). When 14 patients with heterozygous FH and arterial lesions were compared to another 14 without lesions and matched for age and gender, it was found that patients with lesions had on average lower concentrations of HDL-cholesterol, and that 10 of 14 cases were actual smokers. PMID- 3041983 TI - [Breast carcinomas and the extracellular matrix]. AB - This paper underlines the interrelations between tumoral cells and the extra cellular matrix in breast cancers with possible applications for the diagnosis and the prognosis. In mammary carcinomas, the first step of tumoral invasion is characterized by the loss of basement membrane components, particularly type IV collagen and laminin. Immunohistochemical detection of these disruptions of basement membrane is easy and useful for the diagnosis of "in situ" or microinvasive carcinomas. Laminin seems also to facilitate the adhesion of cancer cells to type IV collagen, and the dosage of its fragment P1 in the blood serum may be a good marker for the follow up of the patients. Stromal reaction involves many intricate macromolecules of the extra-cellular matrix. Types I and III collagens are often present in non colloid carcinomas. Rate, turn over of elastin and its prognostic value are still debated. Elastosis is related to well differentiated carcinomas and the presence of estrogen receptors. The stroma of the colloid form of breast cancer is rich in proteoglycans. Malignant and stromal cells, through the intermediary of cytokines, can synthesize these macromolecules. Hyaluronic acid and chondroitin sulfate are abundant in mammary carcinomas and form a favorable substrate for the growth and the migration of malignant cells. However, proteases decrease and limit their action. The presence of fibronectin, principally in the stroma, is difficult to interpret but fibronectin seems to play a role in tumoral retraction. PMID- 3041985 TI - [Optimizing anesthesia ventilation. 2. Anesthesia ventilation with intermittent positive pressure ventilation]. PMID- 3041984 TI - [Pathological anatomy of adenocarcinoma of the prostate in 1988]. AB - After going through gross and microscopic features of prostatic adenocarcinoma, the author stresses upon cytological diagnosis on fine-needle (Franzen) aspiration biopsies. Histological or cytological diagnosis may be difficult. And prostatic dysplasia is an especially difficult subject. There are too many definitions of Stage A1 adeno-carcinoma of the prostate. One would be enough! Histoprognosis only has a statistical value. The Gleason classification is the one we would choose. Any other system could be used as a second classification. PSA (prostate specific antigen) is the best marker for the prostate and for prostatic adenocarcinoma. The idea of an "endocrine prostate" is a new contribution for a better histoprognosis. Carcinoid tumor is only one extreme part of the subject. New methods are reviewed, which will probably make diagnosis and prognosis much easier. PMID- 3041986 TI - [General review: classification of urticaria and angioedema]. AB - The authors propose a classification of urticarias and angio-oedemas according to two criteria, physiopathological and etiological. In the first group they identify three subgroups; immunological, non-immunological and idiopathological. In the second group they classify the facts into 12 sub-groups, which are detailed. PMID- 3041988 TI - [Eczema and hyper-IgE. Effect of cephalosporins]. AB - Atopic dermatitis is a disease in which staphylococcal cutaneous superinfection and immune disorders, especially hyper IgE, are closely related. Cephalosporins are very effective on exacerbations of superinfections; among them, Cefadroxil diminishes immunoglobulins E in vivo. In vitro, this antibiotic promotes from the macrophage the PgE2 synthesis to which the T suppression is classically sensitive. In DNP-OVA sensitized rats which produce anti DNP IgE, the secondary response is inhibited by Cefadroxil. From clinical to research experiments, the hypothesis is the effect of cephalosporins, and especially Cefadroxil, on IgE synthesis. PMID- 3041987 TI - [The G.E.R.U. retrospective study of 400 cases of urticaria]. AB - The authors report on a study with computerised results of 400 urticaria and/or angioedema cases, which were explored in single dermatology center (Pr. J. SAYAG) by the members of GERU. The study was managed with a computerised file and was processed with Sesame software. The figures achieved are related to semilogic and paraclinical data and to etiological factors. PMID- 3041989 TI - Comparison of reaction rates in trypsin-catalysed transamidation of porcine insulin and its B29-arginine analogue. AB - [B29-Arginine]porcine insulin was prepared from des-(B23-30)-insulin and synthetic octapeptide with the aid of trypsin. Comparison of reaction rates in trypsin-catalysed transamidation of this compound and porcine insulin with threonine ether ester showed that this reaction is determined only by conformational effects and structural features of amino acids leaving from and entering into B30, not by the structure and the pKa value of the basic amino acid in B29. PMID- 3041990 TI - Continuous flow peptide synthesis on aminopolyoxyethylene-polystyrene graft copolymer using Fmoc-strategy. AB - An insoluble graft copolymer consisting of the covalently bound polyoxyethylene to cross-linked polystyrene (HO-POE-PS) was prepared by anionic polymerization of ethylene oxide on the resin. The copolymer was then converted to the corresponding amino-polymer (H2N-POE-PS) and the latter was employed as the solid carrier for peptide synthesis. Although HO-POE-PS has successfully been employed as a carrier for peptide synthesis by the standard shaking procedure using t butoxycarbonyl-amino acids, now we deemed it of interest to test its suitability for the continuous flow synthesis. Thus, the C-terminal octapeptide of the porcine insulin B chain (B23-30) was prepared by this procedure using a photolabile anchoring group and fluoren-9-ylmethoxycarbonyl-amino acids. All the reactions were carried out in a continuous flow manner in a steel column under pressure using a high-performance liquid chromatography (HPLC) system. At the end of the synthesis, a sample of the protected peptide was cleaved from the support by photolysis. For the cleavage of another sample, an aqueous solution of sodium carbonate was employed. The protected peptide was purified on silica gel and Sephadex-LH 20. All the protecting groups of a sample of the octapeptide were removed with piperidine/dimethylformamide and trifluoroacetic acid and the deblocked peptide was purified by ion-exchange chromatography. The free peptide was shown to be homogeneous by thin-layer chromatography, HPLC, and electrophoresis. The identify of the free octapeptide was confirmed by amino-acid analysis, 13C-nuclear magnetic resonance measurement and field-desorption mass spectrometry. The peptide was also shown to be free of racemization. PMID- 3041991 TI - [An immunoenzyme method of diagnosing familial hypercholesterolemia]. AB - A new modification of enzyme immunoassay: enzyme-linked-immunoreceptor assay (ELIRA)--was used to study the activity of LDL-receptors on cultured fibroblasts from 10 patients with elevated plasma cholesterol levels, IHD, accelerated atherosclerosis and xanthomatosis. Four patients were found to have heterozygous form of familial hypercholesterolemia. We have also shown that the results of ELIRA were quantitatively similar to the data obtained by traditional radioisotopic method. This indicates that simple, rapid, inexpensive ELIRA can be used for diagnosis of FH. PMID- 3041992 TI - [Inactive renin: its activation, biochemical properties and role in the physiological effects of the renin-angiotensin system]. PMID- 3041993 TI - [Calcium antagonists: the mechanism of their action and aspects of their clinical use in cardiac insufficiency]. PMID- 3041994 TI - [Severe and malignant arterial hypertension: the optimal treatment]. AB - 54 patients were treated with a combination of capoten and diuretics for 40 to 250 days. Some patients had been subjected to the "nitroprusside test" prior to medication. The relation between the clinical state, the perilimbal and eye fundus microcirculation and degree of BP decrease were studied using fluorescent angiography. An adequate (for this group of patients) effect was observed when blood pressure decreased by 25-30% as compared to the initial level. For this capoten, up to 150 mg per day, is sufficient. However, when the BP during the "nitroprusside test" and capoten "overdosage" fell by 35-40% or more as compared to the basal level, undesirable changes were observed. The angioarchitecture deteriorated considerably due to a significant increase in the number of non perfused vessels and vast ischemic zones as compared to the initial state. PMID- 3041995 TI - [Effect of captopril on insulin secretion and blood glucose in patients with arterial hypertension]. AB - In 16 hypertensive patients levels of glucose, immunoreactive insulin and C peptide were studied during oral glucose tolerance test before and 10 days after treatment with captopril. The analysis of the results obtained showed that captopril had no effect on carbohydrate metabolism which is advantageous for treatment of hypertensive patients. PMID- 3041996 TI - Cardiovascular effects of neuropeptide Y. AB - Neuropeptide Y (NPY) is present in the brain, the adrenal medulla, and peripheral sympathetic nerves. This peptide is released together with catecholamines during sympathoadrenal activation. It possesses direct vasoconstrictor properties that are not dependent on simultaneous adrenergic activation. Moreover, it potentiates the vascular effect of several stimulatory substances and may contribute to the modulation of blood pressure responsiveness under a number of circumstances. NPY may also be indirectly involved in the control of blood pressure through regulating the release of hormones with well-established actions on the cardiovascular system. PMID- 3041997 TI - Diabetic nephropathy. The basis for dietary and converting enzyme inhibitor therapy. AB - Once initiated, renal disease progresses in most patients to end-stage over a matter of months or years. This progressive consumption of renal mass seems to result from maladaptations to the initial insult by the remaining kidney. This review stresses the hemodynamic underpinnings of this progression of renal disease. The evidence is reviewed that indicates that loss of renal mass from surgical reduction in kidney tissue or from diabetes mellitus results in increased blood flow, filtration, and glomerular pressure in the remaining nephrons. Prevention of this glomerular hypertension by reduction in dietary protein or by the addition of converting enzyme inhibitors affords protection. The clinical implications of these observations are reviewed. PMID- 3041998 TI - Electrophysiologic evaluation of the facial nerve in Bell's palsy. A review. AB - Facial nerve paralysis is the most common mononeuropathy and idiopathic facial paralysis (Bell's palsy) the most common seventh nerve disease electromyographers may be asked to evaluate. The electrophysiologic method of choice to assess the facial nerve is side-to-side evoked amplitude comparison with the affected side expressed as a percentage of the nonaffected side. This examination should be performed on days 3, 5, 7, 9, 11 and 13 after onset of paralysis. If the percentage of surviving axons falls below 10% within the first 14 days, an incomplete recovery is suggested. Electromyography may assist in prognosticating a functional return, determining neural conduction across the site of injury and following reinervation in the recovery period. The persistence or early return of an absent R1 component of the blink reflex may qualitatively suggest a satisfactory functional outcome in facial paralysis. Supramaximally exciting the facial nerve at the stylomastoid foramen and comparing the clinical response on the affected and nonaffected side, maximum stimulation test, can also predict eventual seventh nerve return. Observing a minimal twitch, utilizing the nerve excitability test or measuring the facial nerve latency have yielded poor correlations with functional return and are of limited usefulness in the prognostication of acute facial palsies. Trigeminal somatosensory evoked potentials can be employed to evaluate the status of the trigeminal nerve as approximately 50% of patients with Bell's palsy also have lesions involving the fifth nerve. Side-to-side amplitude comparison and electromyography are the two most valuable electrophysiologic methods of assessing facial nerve functioning. PMID- 3041999 TI - Threshold perimetry and the diagnosis of glaucoma. AB - Visual field examinations are one of the commonest, presumably most sensitive practical indices of optic nerve dysfunction in glaucoma. Given the difficulties in training and retaining good technicians, and presumed vagaries of the Goldmann type isopter examination technique, interest has turned to computer-controlled automated suprathreshold and threshold static perimetry. Despite the enthusiasm for these approaches, we lack rigorous tests of their value, criteria of abnormality and progressive deterioration, and even normal standards. These important issues are now being addressed. In the interim ophthalmologists must be cautious in their use and interpretation. PMID- 3042001 TI - Pseudosarcomatous lesions and 'borderline' tumors of soft tissues. Historical review. PMID- 3042000 TI - Avoidable blindness. AB - The vast majority of the world's 42 million blind are needlessly impaired. Epidemiologic studies are providing important insights into the cause of cataracts and provision of surgical services; ecologic approaches to the control of trachomatous corneal scarring; treatment and prevention of onchocerciasis; and early diagnosis and treatment of xerophthalmia among others. Continued research and application of existing knowledge can have a dramatic impact on the sight and lives of millions of people. PMID- 3042002 TI - Extracardiac rhabdomyoma: An immunocytochemical study and review of the literature. AB - Two cases of 'adult' rhabdomyoma and 2 of 'fetal' rhabdomyoma have been immunologically studied using several antisera specific of skeletal muscle fibers proteins. This paper also deals with a review of the literature on these rare benign skeletal muscle proliferations. It is concluded that 'adult' rhabdomyoma contains fetal myosin and therefore the term 'adult' does not seem immunologically appropriate any longer. In addition the tumoral cells show the same level of differentiation as seen in neonatal skeletal muscle. Therefore it seems that the definition of neonatal rhabdomyoma is more appropriate for these benign muscular proliferations. The 2 cases of fetal rhabdomyoma appear phenotypically similar to normally developing myoblasts and contain fetal myosin. Therefore it seems that the definition 'fetal' is here appropriate for what appears to be a disorganized proliferation of otherwise normal fetal skeletal muscle fibers. The differential diagnosis between these latter tumors and rhabdomyosarcomas has been discussed. PMID- 3042003 TI - Cartilaginous tumours of soft tissue. AB - Tumours of soft tissue may manifest cartilaginous differentiation as a primary phenomenon (as in chondromas or extraskeletal myxoid and mesenchymal chondrosarcomas) or as a secondary metaplastic feature (most notably in extraskeletal osteosarcomas or malignant nerve sheath tumours). The literature regarding primary cartilaginous tumours is reviewed and their differential diagnosis discussed. Soft tissue chondromas are noteworthy for their tendency to show significant nuclear pleomorphism. Extraskeletal chondrosarcoma is much rarer than its osseous counterpart, accounting for only 1-2% of soft tissue sarcomas. Virtually all such sarcomas can be classified into the myxoid or mesenchymal subtypes, of which myxoid carries a better prognosis. Soft tissue neoplasms which may show focal cartilaginous metaplasia are covered more briefly. PMID- 3042004 TI - [Characteristics of ischemic cerebrovascular disease in Japanese--a review]. PMID- 3042005 TI - [Intravenous hydrocortisone in large doses in the treatment of delayed ischemic neurological deficits following subarachnoid hemorrhage--results of a multi center controlled double-blind clinical study]. AB - Glucocorticoid in high dose has known to reduce vascular sensitivity to various vasoconstrictive stimuli. It inhibits phospholipase to reduce production of prostaglandins. It stabilizes the cell membrane and prevents cerebral edema. All these pharmacological effects indicate possible usefulness of this drug for the treatment of cerebral ischemia due to vasospasm. Based on these theoretical backgrounds a multi-center controlled double blind clinical study was carried out. The patient who showed manifestations of delayed cerebral ischemia due to vasospasm were selected for this study. As soon as the clinical manifestation appeared, the patient was given either 3 grams of hydrocortisone intravenously in a 60 ml solution or the placebo. The administration was repeated 6 times with interval of 12 hours. The patients were allowed to be treated according to the independent protocol of each institute except for the maximum daily use of glucocorticoid. The effect of the therapy was evaluated at 4th, 7th day and 1 month after the initiation. The study involved 52 institutes and a total of 140 patients, seventy-one patients who received hydrocortisone (group A) and 69 patients who received placebo (group P) was analysed. There were no significant differences in background data between both groups. In patients with grade I, II or III on admission, the favorable effects of hydrocortisone were demonstrated on changes in neurological findings. In group A, there were significantly more cases of improvement at 4th day for orientation about place and person. At 1 month or on discharge group A showed significant improvement almost in every aspect of neurological findings including mental, speech and motor functions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3042007 TI - Badges of the dental profession. BDA East of Scotland Branch. PMID- 3042006 TI - New materials, bonding treatments and changes in restorative practice. PMID- 3042008 TI - The structure of fucosylated blood group substances in fetal rat skin. The combined use of monoclonal antibodies and glycosyl hydrolases. AB - Cell surface carbohydrates are thought to be important during embryonic development. We have studied a group of defined cell surface carbohydrates during the differentiation of fetal rat skin by the use of immunocytochemical techniques. We used monoclonal antibodies to the LeX and human blood group H type II haptens. The structure of the molecules carrying these haptens has been inferred by the sensitivity of the immune reactions to digestion by glycosyl hydrolases. The primitive ectoderm expressed the LeX hapten. The differentiating epidermis carries the H type II antigen, but this was only detected following cleavage of a terminally modified portion of the carbohydrate containing linear poly-N-acetyl lactosamine chains. These carbohydrates on epidermal cells are carried N linked to glycoproteins. PMID- 3042009 TI - Effect of UVB therapy and a coal tar bath on short contact dithranol treatment for psoriasis. AB - The effect of UVB therapy after short contact dithranol therapy (SCDT) was examined in 53 patients with psoriasis. After dithranol application, patients were randomly allocated to receive either a tar bath and UVB therapy (27 patients) or an emulsifying ointment bath only (26 patients). Forty eight patients completed the study; 16 of the 21 dithranol only patients cleared in a mean of 19.5 days compared with 20 of the 27 dithranol + UVB patients who cleared in a mean of 20.3 days. These differences were not significant. Similarly there was no significant difference in response between the dithranol only and dithranol UVB group as measured by the change in plaque thickness measured using Harpenden calipers, and the severity of psoriasis assessed by the extent of the rash, the degree of scaling, redness and induration. Thirteen of the 16 dithranol only patients who cleared relapsed in a mean of 10.6 weeks compared with 14 of the 20 dithranol UVB patients who relapsed after a mean of 18.9 weeks (P less than 0.05). Comparison of pre- and post-treatment full blood count, biochemical screen and urinalysis showed no evidence of systemic toxicity due to SCDT. This study shows that UVB therapy does not improve the clearance of psoriasis in SCDT, but does significantly postpone relapse. PMID- 3042010 TI - Objective assessments of port wine stains: response to temperature change. AB - Three objective measurements were made in 11 patients with facial port wine stains and in five controls. Laser Doppler flowmetry, reflectance spectrophotometry and skin surface temperature were all significantly elevated in the port wine stains compared with the normal contralateral cheeks and control skin (P less than 0.05). Changes in skin blood flow and temperature after local heating and cooling of the skin were similar at all sites assessed and were not associated with significant changes in reflectance spectrophotometry, the parameter which correlated most closely with skin colour. PMID- 3042012 TI - Defining dependence. AB - The need to define dependence arises out of accumulating evidence that it contributes to depression-proneness. Its accurate definition is therefore a prerequisite for the development of valid methods of measuring it. Because dependence is more a feature of childhood than of adulthood, a dependent person may usefully be viewed as an adult behaving as though he were a child. Dependence is best understood therefore in terms of those developmental deficiencies from which it results. These are failure to separate successfully from the principal parent figure and from the family as a whole, failure to establish a secure personal identity, failure to acquire a general feeling of competence and a realistic assessment of self-worth and failure to feel deserving of the status of adult and to feel on equal terms with other adults. Consequently, adult dependence is characterized by the need to stay close to others, the inclination to be primarily the recipient in interpersonal transactions and the tendency to relate to others from a position of inferiority and humility. The dependent person receives from others a borrowed identity, guidance and direction, compensation for those areas in which he is incompetent and, most important of all, acceptance, approval and affirmation of worth. PMID- 3042011 TI - Treatment of mycosis fungoides with recombinant interferon-alpha 2a2 alone and in combination with etretinate. AB - Eleven patients with mycosis fungoides (MF) were treated with recombinant alpha interferon (rIFN-alpha 2a2) in combination with etretinate (seven patients) or alone. One patient, who also received etretinate, went into complete remission and remained without signs of MF after 18 months. Six patients experienced partial remission; one of these was treated with rIFN-alpha 2a alone and was clinically in complete remission, but had still a pleomorphic skin infiltrate. Two patients were non-evaluable, and two stopped therapy due to progressive disease. Five patients discontinued therapy due to side-effects although three had partial remission of their disease. Only four patients received 12 months therapy. The study shows that rIFN-alpha 2a in combination with etretinate or alone can induce remission of MF. PMID- 3042013 TI - Women doctors for women patients? AB - This review concentrates on two main areas: (i) the attitudes of women patients towards physicians and (ii) the attitudes of physicians towards female patients. Women are important users of the health care services. There is female to male ratio of 3.8:3.0 for the average number of consultations in general practice per person per year. Women are prescribed more drugs than men; in 1974 66 per cent of women and 54 per cent of men had at least one drug prescribed. Physician gender is thought to have an effect through the physician-patient relationship and its outcomes. Patients perceive male and female doctors differently. Women patients believe women doctors to have the good qualities of both male and female physicians, e.g. assertiveness and initiative but also tenderness and nurturance. There is some evidence that women doctors give their female patients more surgery time than is allocated to them by male doctors. In the specialty areas, preference for a female gynaecologist is stated more often than a preference for a female doctor in any other specialty. The attitudes of physicians of both sexes towards female patients is seen by some as negative; however, physicians as a group are more 'feminist' than control groups. Some women GPs are seeing up to 85 per cent female patients. PMID- 3042014 TI - Video display terminals and pregnancy. A review. PMID- 3042015 TI - Suspected ectopic pregnancy: ultrasound findings and hCG levels assessed by an immunofluorometric assay. AB - One hundred suspected ectopic pregnancies were assessed by ultrasound on the basis of the following criteria: (A) viable intrauterine fetus, intrauterine pregnancy is certain; (B) intrauterine double sac or eccentric ring, intrauterine pregnancy is probable; (C) empty uterus or central ring but no adnexal mass or cul-de-sac fluid, ectopic pregnancy is possible; (D) empty uterus or central ring and an adnexal mass or cul-de-sac fluid, ectopic pregnancy is probable; (E) viable ectopic fetus, ectopic pregnancy is certain. Serum human chorionic gonadotrophin (S-hCG) was detected by an immunofluorometric assay (sensitivity 0.2 i.u./l, cut-off level 10 i.u./l). All the 51 patients in groups A and B had an intrauterine pregnancy. Normal gestational sacs were found also at S-hCG levels of less than 3600 i.u./l, the lowest level being 894 i.u./l. Ectopic pregnancy was confirmed in 29 of the 30 women in groups D and E. In the 19 women categorized into group C serial hCG assay and repeated sonography diagnosed ectopic pregnancy in 12 and miscarriage of an intrauterine pregnancy in the other seven. Ectopic pregnancy was always found when no gestational sac was seen by sonography and the hCG level was greater than 1000 i.u./l. PMID- 3042016 TI - Patterns of growth of uterine leiomyomas during pregnancy. A prospective longitudinal study. AB - In a prospective study 32 leiomyomas (fibroids) in 29 pregnant women were examined with ultrasound every 3-8 weeks. Each patient had between 3 and 6 scans (mean 4.4) during the course of pregnancy, and 13 patients had a final scan at 6 weeks postpartum. An individual growth curve was established for each tumour and the patterns of growth were analysed. No increase in size during the pregnancy was observed in 25 fibroids (78%). Only 7 (22%) increased in size but by no more than 25% of the initial volume. At 6 weeks postpartum the size of the fibroids did not differ significantly from the size during pregnancy. PMID- 3042017 TI - Adenomatoid tumour of uterus. Case report. PMID- 3042018 TI - Mucinous cystadenocarcinoma in pregnancy. Case report. PMID- 3042020 TI - Commentary: The safety of diagnostic ultrasound. PMID- 3042019 TI - Recovery of the intrauterine contraceptive device from the sigmoid colon. Three case reports. PMID- 3042021 TI - Cytotoxic T lymphocyte mediated cytolysis. PMID- 3042022 TI - Structure and dynamics of a detergent-solubilized membrane protein: measurement of amide hydrogen exchange rates in M13 coat protein by 1H NMR spectroscopy. AB - The coat protein of bacteriophage M13 is inserted into the inner membrane of Escherichia coli where it exists as an integral membrane protein during the reproductive cycle of the phage. The protein sequence consists of a highly hydrophobic 19-residue central segment flanked by an acidic 20-residue N-terminus and a basic 11-residue C-terminus. We have measured backbone amide hydrogen exchange of the protein solubilized in perdeuteriated sodium dodecyl sulfate using 1H nuclear magnetic resonance (NMR) spectroscopy. Direct proton exchange out measurements in D2O at 24 degrees C were used to follow the exchange of the slowest amides in the protein. Multiple exponential fitting of the exchange data showed that these amides (29 +/- 3 at pH 4.5) exchanged in two kinetic sets with exchange rates [(1.2 +/- 0.4) x 10(-4) s-1 and (4.1 +/- 1.2) x 10(-7) s-1] that differed by more than 100-fold, the slower kinetic set being retarded 10(5)-fold relative to poly(DL-alanine). The exchange rate constant for the slowest set of amides exhibited an unusual pD dependence, being proportional to [OD-]1/2. It is shown that this is an artifact of the multiple exponential fitting of the data, and a new method of presentation of exchange data as a function of pD is introduced. Steady-state saturation-transfer techniques were also used to measure exchange. These methods showed that 15-20 amides in the protein are very stable at 55 degrees C and that about 30 amides have exchange rates retarded by at least 10(5)-fold at 24 degrees C. Saturation-transfer studies also showed that the pH dependence of exchange in the hydrophilic termini was unusual. This is explained as being due to long-range electrostatic effects arising both from the protein itself and also from the anionic detergent molecules. Hydrogen exchange studies on the products of proteinase K digestion of the protein localized the slowly exchanging amides to the hydrophobic core of the protein. Relaxation [Henry, G.D., Weiner, J.H., & Sykes, B.D. (1986) Biochemistry 25, 590-598] and solid state NMR experiments [Leo, G.C., Colnago, L.A., Valentine, K.G., & Opella, S.J. (1987) Biochemistry 26, 854-862] have previously shown that the majority of the protein backbone is rigid on the picosecond to microsecond time scale, except for the extreme ends of the molecule which are mobile.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3042024 TI - Expression of active rat DNA polymerase beta in Escherichia coli. AB - A recombinant plasmid for expression of rat DNA polymerase beta was constructed in a plasmid/phage chimeric vector, pUC118, by an oligonucleotide-directed mutagenesis technique. The insert contained a 1005 bp coding sequence for the whole rat DNA polymerase beta. The recombinant plasmid was designed to use the regulatory sequence of Escherichia coli lac operon and the initiation ATG codon for beta-galactosidase as those for DNA polymerase beta. The recombinant clone, JMp beta 5, obtained by transfection of E. coli JM109 with the plasmid, produced high levels of DNA polymerase activity and a 40-kDa polypeptide that were not detected in JM109 cell extract. Inducing this recombinant E. coli with isopropyl beta-thiogalactopyranoside (IPTG) yielded amounts of 40-kDa polypeptide as high as 19.3% of total protein. Another recombinant clone, JMp beta 2-1, which was constructed by an oligonucleotide-directed mutagenesis to use the second ATG codon for the initiation codon, thus deleting the first 17 amino acid residues from the amino terminus, produced neither high DNA polymerase activity nor the 40 kDa polypeptide. The evidence suggests that this amino-terminal structure is important for stability of this enzyme in E. coli. The DNA polymerase was purified to homogeneity from the IPTG-induced JMp beta 5 cells by fewer steps than the procedure for purification of DNA polymerase beta from animal cells. The properties of this enzyme in activity, chromatographic behavior, size, antigenicity, and also lack of associated nuclease activity were indistinguishable from those of DNA polymerase beta purified from rat cells, indicating the identity of the overproduced DNA polymerase in the JMp beta 5 and the rat DNA polymerase beta. PMID- 3042023 TI - Amino acid sequence of bovine cardiac troponin I. AB - Troponin I (TnI) is the inhibitory subunit of troponin, the thin filament regulatory complex which confers calcium sensitivity to striated muscle actomyosin ATPase activity. We have determined the amino acid sequence of TnI from adult bovine cardiac muscle. This protein is a single polypeptide chain of 211 amino acids with an acetylated amino terminus, a calculated molecular weight of 23,975, and a net charge of +17 at neutral pH. There was no evidence for heterogeneity of the sequence. Comparison with other available TnI sequences shows an amino-terminal extension of 27-33 residues which is present in cardiac but not skeletal TnI. The remainder of the polypeptide is common to both cardiac and skeletal TnI. In the amino-terminal half of the common polypeptide, only 29% of the residues are invariant in all sequences. The carboxyl-terminal half (residues 124-210) is much more highly conserved, with 66% invariant residues. Bovine cardiac TnI and rabbit cardiac TnI are very similar in sequence: only 12 of 26 residues are identical in the amino-terminal segments, but the remaining residues of the proteins are 97% identical. PMID- 3042025 TI - Furosemide and Ca2+ affect 86Rb+ efflux from pancreatic beta-cells by different mechanisms. AB - The interaction between furosemide, calcium and D-glucose on the 86Rb+ efflux from beta-cell-rich mouse pancreatic islets was investigated in a perifusion system with high temporal resolution. Raising the glucose concentration from 4 to 20 mM induced an initial decrease in 86Rb+ efflux, which was followed by a steep increase and then a secondary decrease. Removal of extracellular calcium increased the 86Rb+ efflux at 4 mM D-glucose but reduced it at 20 mM. The initial biphasic changes in 86Rb+ efflux induced by 20 mM D-glucose were inhibited by calcium deficiency. Furosemide (100 microM) reduced the 86Rb+ efflux rate both at 4 and 20 mM D-glucose and the magnitudes appeared to be similar at either glucose concentration. Furosemide (100 microM) reduced the glucose-induced (10 mM) 45Ca+ uptake but did not affect the basal (3 mM D-glucose) 45Ca+ uptake. However, the ability of furosemide (100 microM) to reduce the 86Rb+ efflux at a high glucose concentration (20 mM) was independent of extracellular calcium. The inhibitory effects of furosemide and calcium deficiency on the 86Rb+ efflux rate appeared to be additive. It is concluded that the effect of furosemide on 86Rb+ efflux is not secondary to reduced calcium uptake and that the effects of furosemide and calcium deficiency are mediated by different mechanisms. The effect of furosemide is compatible with inhibition of loop diuretic-sensitive co-transport of Na+, K+ and Cl- and the effect of calcium deficiency with reduced activity of calcium regulated potassium channels. PMID- 3042026 TI - The apical and basal plasma membranes of the human placental syncytiotrophoblast contain different erythrocyte membrane protein isoforms. Evidence for placental forms of band 3 and spectrin. AB - Using immunochemical techniques, we identified forms of erythrocyte membrane proteins in apical and basal plasma membranes of human placental trophoblast. A wheat germ agglutinin-binding intrinsic protein was present in the microvillous (maternal facing) but not the basal (fetal facing) membrane of the syncytiotrophoblast epithelium. Conversely, erythrocyte-related proteins of the basal membrane included two intrinsic membrane proteins, a 95,000 Mr band 3 isoform and a form of spectrin. These four proteins were all absent from the microvillous membrane. The basal membrane spectrin isoform was also present in basal membrane skeletons. A 70,000 Mr polypeptide which reacted with antibodies to band 3 was present in both microvillous and basal plasma membranes. Therefore, certain isoforms of red cell membrane proteins are polarized between the two surfaces of the human placental syncytiotrophoblast. We propose that the localization of spectrin to the basal membrane is related to the less bundled organization of microfilaments at this membrane compared with that of the microvillous membrane. The band 3 isoforms are candidates for participation in maternofetal anion transport. PMID- 3042027 TI - Synergistic binding of glucose and aluminium ATP to hexokinase from Saccharomyces cerevisiae. AB - The binding of glucose, AlATP and AlADP to the monomeric and dimeric forms of the native yeast hexokinase PII isoenzyme and to the proteolytically modified SII monomeric form was monitored at pH 6.7 by the concomitant quenching of intrinsic protein fluorescence. No fluorescence changes were observed when free enzyme was mixed with AlATP at concentrations up to 7500 microM. In the presence of saturating concentrations of glucose, the maximal quenching of fluorescence induced by AlATP was between 1.5 and 3.5% depending on species, and the average value of [L]0.5, the concentration of ligand at half-saturation, over all monomeric species was 0.9 +/- 0.4 microM. The presence of saturating concentrations of AlATP diminished [L]0.5 for glucose binding by between 260- and 670-fold for hexokinase PII and SII monomers, respectively (dependent on the ionic strength), and by almost 4000-fold for PII dimer. The data demonstrate extremely strong synergistic interactions in the binding of glucose and AlATP to yeast hexokinase, arising as a consequence of conformational changes in the free enzyme induced by glucose and in enzyme-glucose complex induced by AlATP. The synergistic interactions of glucose and AlATP are related to their kinetic synergism and to the ability of AlATP to act as a powerful inhibitor of the hexokinase reaction. PMID- 3042028 TI - Lipid and lipoprotein metabolism in Hep G2 cells. AB - Lipid composition, lipid synthesis and lipoprotein secretion by the Hep G2 cell line have been studied with substrate and insulin supplied under different conditions. The lipid composition of Hep G2 cells was close to that of normal human liver, except for a higher content in sphingomyelin (P less than 0.005) and a lower phosphatidylcholine/sphingomyelin ratio. Most of the [14C]triacylglycerols secreted into the medium were recovered by ultracentrifugation at densities of 1.006 to 1.020 g/ml. The main apolipoproteins secreted were apo B-100 and apo A-I. Hep G2 mRNA synthesized in vitro the pro apolipoproteins A-I and E. Triacylglycerol secretion was 7.38 +/- 1.04 micrograms/mg cell protein per 20 h with 5.5 mM glucose in the medium and increased linearly with glucose concentration. Oleic acid (1 mM) increased the incorporation of [3H]glycerol into the medium and cell triacylglycerols by 251 and 899%, with a concomitant increment in cell triacylglycerols and cholesterol ester. Insulin (1 mU or 7 pmol/ml) inhibited triacylglycerol secretion and [35S]methionine incorporation into secreted protein by 47 and 28%, respectively, with a corresponding increase in the cells. Preincubation of cells with 2.5-10 mM mevalonolactone decreased the incorporation of [14C]acetate into cholesterol 6.2 fold, indicating an inhibitory effect on HMG-CoA reductase. It is concluded that in spite of some differences between Hep G2 and normal human hepatocytes, this line offers an alternative and reliable model for studies on liver lipid metabolism. PMID- 3042029 TI - Purification and characterization of a novel cytotoxic substance from cell-free extract of Streptococcus pyogenes. AB - A cytotoxic substance designated as streptococcal cytotoxic protein (SCP) was isolated from a cell-free extract of the Su strain of Streptococcus pyogenes possessing cytotoxic and antitumor activity. SCP was purified with a series of column chromatography and preparative PAGE to give a homogeneous single band as revealed by PAGE analysis. The purified SCP has a molecular mass of 165 kDa, composed of four 43 kDa subunits, and its pI is 4.3. SCP was sensitive to proteinases and was labile to heat and at acidic or alkaline pH. SCP showed inhibitory effects on the [3H]thymidine, [3H]uridine and [3H]leucine uptakes and on the growth of cells, and released 51Cr from cells when the protein was added to the cultures of Ehrlich ascites carcinoma (EAC), mouse mammary tumor (MM-2), leukemia (L-1210) and NIH-3T3 mammalian cells in vitro. SCP also showed an antitumor effect on EAC or MM-2 tumor-bearing mice but not on L-1210 tumor bearing mice in vivo. PMID- 3042030 TI - The ultrastructure of A, B and D pancreatic cells in normal and in diabetic ducks. AB - Ultrastructurally and immunocytochemically identified A, B and D cells are highly concentrated in the splenic bulb of the duck pancreas. Ultrastructural features of normal A, B and D cells are similar in the duck and in other species so far studied. However, normal D cells present a striking characteristic, i.e. apical accumulation of dense bodies, which seem to derive from multivesicular bodies and are probably involved in a catabolic regulatory process. Subtotal pancreatectomy in the duck, leaving the splenic bulb and inducing transient diabetes, produces strong secretory stimulation of A and B cells, as indicated by the development of the rough endoplasmic reticulum and the Golgi apparatus and transient degranulation, more marked in B cells. Numerous B cells with degenerative aspects, observed after 12 days, seem to be exhausted following prolonged hyperstimulation: this could explain why diabetes reappears in some cases. In contrast, in D cells, functional inhibition after surgery is suggested by a dramatic increase in the number and size of the dense bodies, associated with a marked decrease in secretory vesicle storage. The morphological data correlate well with the previously reported evolution of plasma and pancreatic hormone concentration after surgery, and suggest that the normal inhibitory control of glucagon and insulin secretion by the local release of somatostatin might be reduced or suppressed during transient diabetes in subtotally depancreatized ducks. PMID- 3042031 TI - Insulin, somatomedin-C, human chorionic gonadotropin, and forskolin enhance glucose oxidation by granulosa cells. AB - The long exposure times required to observe stimulatory effects of insulin on steroidogenesis and protein synthesis in granulosa cells suggested that these effects might be secondary to stimulation of another metabolic process. The present studies examined the effects of insulin, the insulin-like growth factor somatomedin-C (Sm-C), human chorionic gonadotropin (hCG), and forskolin, a compound that activates adenylyl cyclase independently of a receptor, on glucose metabolism. Granulosa cells from preovulatory porcine ovarian follicles were incubated at 37 degrees C in Dulbecco's phosphate-buffered saline supplemented with bovine serum albumin, vitamins, amino acids, and glucose (0.01-20 mM). Cells were incubated with [14C]glucose for up to 23 h with or without a prior 20-h preincubation. Oxidation of glucose, assessed by quantitation of 14CO2 produced, was dependent on time and concentration of glucose. Optimal glucose concentrations for glucose oxidation were 3 mM in the absence or presence of insulin and correlated well with the measured glucose concentrations in follicular fluid (3 mM). After a 20-h preincubation in the absence or presence of insulin (1 microM), the rates of CO2 production were 10.6 and 21.6 pmol/micrograms DNA/h for control and insulin-treated cells, respectively. Insulin had an EC50 of 164 nM. Sm-C and hCG were more potent stimulators than insulin with EC50s of 768 pM and 161 pM, respectively. The greater sensitivity of granulosa cells to Sm-C than to insulin supports the concept that insulin exerts its effect via reactivity with the Sm-C receptor. The effect of hCG may have been mediated by cyclic adenosine 3',5'-monophosphate (cAMP), since forskolin also enhanced 14CO2 production.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3042032 TI - Purification of the guinea pig sperm PH-20 antigen and detection of a site specific endoproteolytic activity in sperm preparations that cleaves PH-20 into two disulfide-linked fragments. AB - Previous work has indicated that the guinea pig sperm membrane protein, PH-20, functions in sperm-egg adhesion and that its surface expression is regulated by the acrosome reaction. The PH-20 protein was purified by monoclonal antibody affinity chromatography. Sixty-seven to one hundred percent of the PH-20 antigenic activity present in an octylglucoside (OG) extract of sperm was recovered in the purified protein. From 10(10) sperm, approximately 0.4 mg of PH 20 protein was obtained, which was about 0.24% of the total protein in the OG extract. The purified protein retained the ability to bind the three anti-PH-20 monoclonal antibodies we have isolated. Silver staining of purified PH-20 on overloaded sodium dodecyl sulfate (SDS) gels allowed the estimate that silver stainable contaminants were present at a level of one part in 2000. The purified PH-20 protein exists in three forms separable on SDS-polyacrylamide gel electrophoresis: a major form with a molecular mass of 64 kDa, a minor form of 56 kDa, and an endoproteolytically cleaved form composed of two disulfide-linked fragments of 41-48 kDa and 27 kDa. Cleveland digests of the 64 kDa and 56 kDa polypeptides indicated that they were structurally related. A proportion of the 64 kDa polypeptide in each purified preparation had undergone endoproteolysis at a specific site, so that it was cleaved into the two disulfide-linked fragments, 41-48 kDa and 27 kDa. It is speculated that the site-specific endoproteolysis of PH-20 may occur during the acrosome reaction and have biological significance. PMID- 3042033 TI - On the means whereby mammals achieve increased functional durability of their dentitions, with special reference to limiting factors. PMID- 3042034 TI - Operation of the purine nucleotide cycle in animal tissues. PMID- 3042035 TI - [Immunochemical identification of beta2-globulin in metastases of ovarian tumors]. AB - Antisera were obtained upon immunization of rabbits with the extracts obtained from metastatic tissues of primary ovarian carcinoma into the omentum. Eight antigens were found, which were termed "ovarian-metastatic antigens" (OMA). OMA 1 7 showed cross-reactions with the antigens of one of the normal organs of the adult man: kidneys, spleen, brain. OMA-8 was not identified in the internal organ tissues of the adults and fetuses. Using immunodiffusion method, OMA-8 was revealed in the tissues of metastases of primary ovarian cancer into the omentum in 55% of cases (3-180 mg/l), in 50% of cases it was detected in primary ovarian carcinomas (3-50 mg/l) and in mature placenta (38-40 weeks) (6-12 mg/l). OMA-8 was detected in the chorion (8-20 weeks) and in the amniotic fluid at all periods at the maximum sensitivity of immunodiffusion method (1-2 mg/l). OMA-8 is a beta 2-globulin of protein nature with NW 35 kD, containing alpha- and beta-subunits with MW 18 and 19.5 kD. No carbohydrates, lipids and ferrum were determined in OMA-8. Its physico-chemical and antigenic properties differ from those of the described carcinoembryonic and placental proteins. PMID- 3042036 TI - [Immunohistochemical study of carcinoembryonic antigen (CEA) in normal and embryonic human tissues using a CEA-specific oncoprecipitin, krustacin]. AB - Using immunoperoxidase technique, antibodies to CEA (Ab) were compared to a glycoprotein krustacin (Kr) extracted from Pagurus prideauxii, which has an ability to precipitate specifically CEA. It was found that Kr and Ab reacted in a similar manner with embryonic and normal gastrointestinal tissues, revealing practically identical localization of the antigen in the tissues and cells. It was possible, however, to note some quantitative and qualitative differences in the distribution of antigen, which showed that Kr and Ab reacted with different determinants of the CEA molecule. PMID- 3042038 TI - Cellular interactions. PMID- 3042037 TI - [Effect of a deficiency in the mononuclear phagocyte system and the administration of yeast polysaccharide on the development of a heterotopic source of hematopoiesis]. AB - Mononuclear phagocyte system (MPS) deficiency was induced by repeated peritoneal lavage in (C57Bl x CBA) F1 mice. The animals were then used as donors or recipients in heterotopic bone marrow transplantation. Yeast polysaccharide (YP) produced by Cryptococcus luteolus strain 228 was injected weekly (25 mg/kg) during 30 days after bone marrow transplantation under the kidney capsule. Bone marrow transplantation from MPC-deficient mice to intact mice 30 days later resulted in no variations from the control in cellularity and ossicle weight. YP produced an increase in cellularity, but not in ossicle weight. In the opposite experimental scheme (transplantation from intact mice to MPS-deficient mice) an increase in both cellularity and weight was not noticed. YP injections in this case resulted in the reduction of heterotopic organ size to the control level. Possible mechanisms of this phenomenon are discussed. PMID- 3042039 TI - Immunoglobulin E levels following allogeneic, autologous, and syngeneic bone marrow transplantation: an indirect association between hyperproduction and acute graft-v-host disease in allogeneic BMT. AB - Markedly elevated serum IgE levels have been noted following allogeneic bone marrow transplantation (BMT) and have been correlated with graft-v-host disease (GVHD) in several studies. To investigate this phenomenon, we measured serum IgE levels in 387 allogeneic, 143 autologous, and 21 syngeneic BMT recipients before and at intervals after BMT. As a population, allogeneic BMT recipients displayed a biphasic elevation in IgE levels, with peak levels occurring either early (days 15 to 19) or late (days 80 to 89) posttransplant. Only in individuals in whom peak levels occurred early did IgE level correlate with liver disease, histological changes, and overall clinical stage of GVHD. The association of IgE elevation and GVHD does not appear to be direct since recipients of syngeneic (monozygotic twin) grafts had the highest incidence of IgE hyperresponsiveness as well as the highest absolute IgE levels. Similarly, 22 recipients of autologous marrow not treated with 4-hydroperoxycyclophosphamide had elevated IgE levels comparable to those seen in allogeneic graft recipients. We hypothesize that augmented IgE synthesis and its subsequent resolution is the natural consequence of immune reconstitution in the presence of potentially reaginic agents such as antibiotics and infectious agents. As such, IgE hyperresponsiveness in syngeneic graft recipients may reflect the maturational sequence of IgE regulatory elements in the absence of interference by GVHD, GVHD therapy, or minor histocompatibility disparities. The cell populations required for IgE response (T cells, B cells, and antigen-presenting cells) may be reconstituted in advance of the regulatory elements that limit IgE production in healthy subjects. Although this temporal relationship does not appear to hold in allogeneic BMT, the balance between positive and negative factors, which determines the rates of IgE synthesis and catabolism, may be altered by GVHD, infection, and liver dysfunction acting alone or in combination. PMID- 3042040 TI - Intravascular hemolysis and renal insufficiency after bone marrow transplantation. AB - Renal disease has not been considered a major late complication of bone marrow transplantation. Of 31 evaluable pediatric patients undergoing allogeneic or autologous bone marrow transplantation for neuroblastoma or acute lymphoblastic leukemia, 14 developed a hemolytic anemia, microscopic hematuria, and renal insufficiency at a median of 5 months (range, 3 to 7 months) posttransplant. Renal biopsies were performed in two patients at the onset of kidney disease and showed mesangiolysis with intraglomerular capillary aneurysm formation, mesangial proliferation, and focal thickening and splitting of the glomerular basement membranes. The clinical presentation, time to onset of renal disease, and biopsy material are consistent with a diagnosis of radiation nephritis, a previously uncommon finding in this patient group. The high incidence of this syndrome in the current report may have been due to the combination of intensive chemotherapy and total-body irradiation in the conditioning regimens. PMID- 3042041 TI - Prednisone and azathioprine compared with prednisone and placebo for treatment of chronic graft-v-host disease: prognostic influence of prolonged thrombocytopenia after allogeneic marrow transplantation. AB - We conducted a randomized, double-blind comparison of prednisone and placebo (group I) v prednisone and azathioprine (1.5 mg/kg/day) (group II) as early treatment of extensive chronic graft-v-host disease (GVHD). Patients with platelet counts less than 100,000/microL were placed into therapy with prednisone alone (group III). All three groups received identical doses of prednisone (1 mg/kg every other day) and one double-strength trimethoprim-sulfamethoxazole (TMP SMX) tablet twice daily. Between January 1980 and December 1983, 179 previously untreated patients were enrolled and 164 were evaluable. Patients randomized to group I (n = 63) and group II (n = 63) were well matched for prognostic factors; those placed into group III (n = 38) had more frequent acute GVHD and progressive onset of chronic GVHD. Median duration of therapy was 2 years. Complications included diabetes (5%), aseptic necrosis (5%) and infection. For groups I, II, and III, the respective incidence of infection was disseminated varicella, 11%, 24%, 34%; bacteremia, 6%, 11%, 34%; and interstitial pneumonia, 5%, 14%, 18%. Recurrent malignancy was the most frequent cause of death and did not differ significantly across the groups. Nonrelapse mortality, however, did differ: 21% in group I, 40% in group II, and 58% in group III (I v II, P = .003; I v III, P = .001). Forty patients in group I, 30 in group II, and 10 in group III survive with a minimum follow-up of 3.8 years. Karnofsky performance scores for 68 survivors are 90% to 100%, scores for seven survivors are 70% to 89% and scores for five survivors are less than 70%. Actuarial survival at 5 years after transplant is 61% in group I, 47% in group II, and 26% in group III (I v II, P = .03; I v III, P = .0001). Treatment with prednisone alone results in fewer infections and better survival than prednisone and azathioprine in standard-risk chronic GVHD. Treatment with prednisone alone is less effective in high-risk patients with thrombocytopenia, and other strategies are required. PMID- 3042043 TI - Granulocyte-macrophage colony-stimulating factor enhances selective effector functions of tissue-derived macrophages. AB - Granulocyte-macrophage colony-stimulating factor (GM-CSF) is produced by a variety of cells at sites of exposure to antigens. GM-CSF has a stimulatory effect on a number of neutrophil functions, but the effect on macrophage function is less clear. We investigated the effect of purified murine recombinant GM-CSF on murine peritoneal macrophage oxidative metabolism, Fc-dependent phagocytosis, anti-Toxoplasma activity, and expression of class II major histocompatibility antigen (Iad). GM-CSF significantly increased phorbol myristate acetate- and zymosan-elicited H2O2 release by resident and thioglycollate-elicited macrophages after 48 hours in vitro. The effect of recombinant GM-CSF was blocked by polyclonal anti-GM-CSF antibody and was not altered by lipopolysaccharide (0.01 to 1.0 microgram/mL). GM-CSF also stimulated Fc-dependent phagocytosis by peritoneal macrophages, although the stimulation of resident macrophages (1.4 fold) was less dramatic than that of thioglycollate-elicited cells (2.1-fold). GM CSF (at doses up to 100 U/mL) had no effect on macrophage anti-Toxoplasma activity or on expression of Iad. In addition to stimulating macrophage growth, GM-CSF selectively promotes the functional capacity of tissue-derived macrophages. PMID- 3042042 TI - Alternating-day cyclosporine and prednisone for treatment of high-risk chronic graft-v-host disease. AB - Therapy of chronic graft-v-host disease (GVHD) has been unsatisfactory in patients with platelet counts less than 100,000/microL. Survival at 5 years after marrow transplant is only 26% in such patients treated with trimethoprim sulfamethoxazole (TMP-SMX) and every other day with prednisone. Since October 1982, 61 patients with high-risk extensive chronic GVHD were treated with a new alternating-day regimen of prednisone (1 mg/kg every other day) and oral cyclosporine (6 mg/kg every 12 hours every other day) with one double-strength TMP-SMX tablet twice daily. Forty patients (group I) received primary treatment of thrombocytopenic chronic GVHD (median platelet count 35 [range 7 to 87] x 10(3)/microL). Twenty-one patients (group II) received salvage treatment after failing initial prednisone +/- azathioprine. Twenty-one patients in group I and 15 in group II survive with a minimum of 2 years and a median of 3.7 years follow up. At 4 years after transplant, actuarial survival is 51% (group I) and 67% (group II). Causes of death included interstitial pneumonia (six), relapse (five), GVHD without infection (five), infection (four), organ failure (three), and hemorrhage (two). Mortality increased with the progressive type onset of chronic GVHD and treatment failure. Toxicity included hypertension (13), nephrotoxicity (nine), nausea (seven), aseptic necrosis (five), neurologic abnormalities (four), and diabetes (three). Median cyclosporine levels at four and 36 hours were 296 and 64 ng/mL, respectively. Four patients required permanent discontinuation of cyclosporine, but none required renal dialysis. Karnofsky performance scores for 25 survivors are 90% to 100%, scores for six survivors are 70% to 89%, and scores for five survivors are less than 70%. Alternating-day cyclosporine and prednisone has acceptable toxicity and appears to improve survival in patients with high-risk chronic GVHD. PMID- 3042044 TI - Bronchiolitis obliterans in bone marrow transplantation and its relationship to chronic graft-v-host disease and low serum IgG. AB - The records of 549 bone marrow transplant (BMT) patients at The Johns Hopkins Oncology Center during a 9-year period were reviewed to determine the incidence of bronchiolitis obliterans (BrOb). Seven patients had BrOb. All seven died, and BrOb was a contributing cause of death in six patients. Only recipients of allogeneic BMT were at risk for developing BrOb (2% incidence). Three cases were incidentally discovered at autopsy in patients who died less than 120 days after BMT from ventilatory failure owing to interstitial pneumonitis. Four cases were patients who died greater than 120 days after BMT. Of this latter group, all had overt chronic graft-v-host disease (CGVHD). Among 120 day survivors of allogeneic BMT, 6% of those with CGVHD developed BrOb as compared with none of those without CGVHD (P = .008). Five percent of patients with reduced IgG levels at day 120 developed BrOb as compared with none of those with normal IgG (P = .04). The incidence of BrOb in 120-day survivors was 14% (4 of 29) in patients with both CGVHD and decreased serum IgG, whereas patients with CGVHD only (0 of 25), those with decreased IgG levels only (0 of 53), and those with no CGVHD and normal IgG levels (0 of 70) did not develop BrOb. PMID- 3042045 TI - Granulocyte-macrophage colony-stimulating factor induces the expression of the CD11b surface adhesion molecule on human granulocytes in vivo. AB - The CD11b (Mol) molecule is a member of a family of surface glycoproteins that are essential for adhesion-dependent granulocyte functions. Brief exposure of granulocytes to human granulocyte-macrophage colony-stimulating factor (GM-CSF) in vitro increases the surface expression of CD11b and increases granulocyte adhesiveness. To assess the possible in vivo significance of these observations we studied the effect of GM-CSF on CD11b, CD11a (LFA-1), and CD11c (gp 150, 95) expression on granulocytes from nine adult patients with sarcoma who were receiving GM-CSF as part of a phase I trial. GM-CSF was administered as a continuous infusion at a dose of 32 or 64 micrograms/kg/d. Granulocyte CD11b, CD11a, and CD11c expression was determined by indirect immunofluorescence staining of whole blood, thereby minimizing in vitro manipulation. A transient leukopenia developed within 15 minutes of initiation of GM-CSF treatment that was associated with a marked increase in the surface antigen density of CD11b. A mean 1.7-fold increase (P = .001) in the percentage of CD11b-positive granulocytes and a mean 2.1-fold increase (P = .002) in CD11b surface antigen density was noted after 12 hours of treatment. No change in CD11a or CD11c expression was observed over the first 12 hours. The level of CD11b expression was followed in six patients for up to 5 days of treatment with GM-CSF. Compared with the 12-hour value, three of six patients showed a subsequent decrease in CD11b expression, two remained constant, and one showed a continued increase in CD11b surface density. Fluorescence-activated cell sorting of granulocytes into high- and low density CD11b-positive groups revealed a preponderance of immature myeloid forms in the low-density CD11b fraction, which suggests that the late decrease in CD11b expression in some patients may be related to a greater proportion of circulating immature myeloid forms in the peripheral blood. This study suggests that GM-CSF administered as a continuous infusion rapidly upregulates the expression of granulocyte CD11b in vivo. The influence of this phenomenon on in vivo granulocyte aggregation may be clinically relevant with regard to the toxicity of GM-CSF and deserves further investigation. PMID- 3042046 TI - Phase I/II study of recombinant human granulocyte-macrophage colony-stimulating factor in aplastic anemia and myelodysplastic syndrome. AB - We performed a phase I/II study of the administration of recombinant human granulocyte-macrophage colony-stimulating factor (GM-CSF) to patients with aplastic anemia or myelodysplastic syndrome. Doses ranging from 15 to 480 micrograms/m2 were administered as a one-hour or four-hour intravenous infusion daily for 7 days or as a 12-hour infusion for 14 days. Temporary improvements were seen in granulocyte counts, monocyte counts, and reticulocyte counts in six of eight patients with aplastic anemia and five of seven patients with myelodysplastic syndromes. The patients with myelodysplastic syndromes had larger increases in granulocyte, monocyte, and reticulocyte counts than did those with aplastic anemia, and they also had increases in the numbers of eosinophils (two of seven), immature myeloid cells (two of seven), and myeloblasts (two of seven) that were not observed in patients with aplastic anemia. There was no reduction in erythrocyte transfusion requirements, and no effect was observed on platelet counts. There was only minimal toxicity consisting of transient low-back discomfort, anorexia, myalgias/arthralgias, and low-grade fever. Our data suggest that GM-CSF is well tolerated and is more likely to result in elevations of blood counts in patients with myelodysplasia than in patients with aplastic anemia, but the role of GM-CSF therapy in these disorders remains to be determined. PMID- 3042047 TI - Synthesis and characterization of an antihuman T-lymphocyte saporin immunotoxin (OKT1-SAP) with in vivo stability into nonhuman primates. AB - The authors conjugated, by a disulphide bond, the antihuman T-lymphocyte (CD5) monoclonal antibody (MoAb) OKT1 to the saporin-6 (SAP) ribosome-inactivating protein of the plant Saponaria officinalis. The resulting OKT1-SAP immunotoxin bound to CD5-expressing target cells and under standard culture conditions specifically suppressed mitogen-induced-T-lymphocyte DNA and protein synthesis in a dose-related manner. T-lymphocyte killing was achieved by five-minute exposure of the target cells to OKT1-SAP. The concentration inhibiting 50% (IC50) of T lymphocyte DNA synthesis was 0.32 nmol/L. The potency of OKT1-SAP was moderately enhanced by amantadine (IC50 0.08 nmol/L) but not by ammonium chloride or chloroquine. Whole blood components did not interfere with the efficacy of OKT1 SAP, as in vitro treatment of fresh whole blood resulted in effective elimination of clonable peripheral blood T-lymphocytes assessed by a limiting dilution assay. Because these characteristics of T-lymphocyte killing by OKT1-SAP (ie, rapidity of action, potency also without potentiators) and lack of inhibition by whole blood components may be relevant for the use of an immunotoxin as a therapeutic agent in humans, the authors evaluated the stability in vivo and the circulatory clearance of OKT1-SAP in cynomolgus monkeys. Following a single intravenous (IV) injection of nontoxic dosages (0.16 to 1.3 mg/kg), an initial rapid decline (t1/2 alpha = 1.0 to 4.1 hours) was followed by a long-lasting slower decrease (t1/2 beta = 11.6 to 20.6 hours) of OKT1-SAP plasma concentrations as detected by double-antibody solid phase enzyme-linked immunosorbent assay (ELISA) assay. Not only did OKT1-SAP remain intact immunologically but it also retained its biological activity, as measured by the ability of plasma samples from monkeys given immunotoxin to inhibit DNA synthesis in human T-lymphocytes. Taken together the findings presented in this article indicate the feasibility of using OKT1-SAP as a therapeutic tool and provide information that will facilitate the rational use of immunotoxins as a treatment modality in humans. PMID- 3042048 TI - Correlated flow cytometric analysis of H-ras p21 and nuclear DNA in multiple myeloma. AB - Correlated analysis of the H-ras oncogenes product (p21) and of nuclear DNA content was performed by flow cytometry (FCM) in patients with DNA-aneuploid multiple myeloma (MM). Bone marrow cells from normal donors and MM patients in remission served as controls. Seventy-four percent of 23 patients with active MM had higher p21 fluorescence in aneuploid tumor cells than were observed in normal donor or myeloma remission bone marrows; 39% of the 23 patients also showed high H-ras p21 expression in diploid cells. There was an inverse relationship between p21 levels and the presence of trisomy 11; especially high p21 levels were noted in patient without trisomy 11. The frequent elevation of p21 protein in aneuploid plasma cells suggests the involvement of the H-ras oncogene in the pathophysiology of MM, which is further supported by a shorter survival among patients with high p21 levels. PMID- 3042049 TI - Interleukin-6 enhances growth factor-dependent proliferation of the blast cells of acute myeloblastic leukemia. AB - The effects of recombinant interleukin-6 (IL-6) on the proliferation of blast precursors present in the peripheral blood of patients with acute myeloblastic leukemia (AML) was investigated. IL-6 had little effect by itself; however, it synergized with granulocyte macrophage colony-stimulating factor (GM-CSF) and interleukin-3 (IL-3) in the stimulation of AML blast colony formation. Responsiveness of blast progenitors to IL-6 was heterogeneous. On normal bone marrow cells the same synergy was observed on granulocyte and monocyte precursors (GM-CFC), while there was no significant effect on erythroid and multipotential precursors. PMID- 3042050 TI - Use of recombinant human granulocyte-macrophage colony-stimulating factor in autologous marrow transplantation for lymphoid malignancies. AB - The use of recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) following autologous marrow transplantation for lymphoid malignancies was explored in a phase I/II dose escalation study. rhGM-CSF given as a 2-hour infusion daily for 14 days was well tolerated at doses up to 240 micrograms/m2/day. When compared with 86 disease-matched and treatment-matched historical controls, patients receiving greater than or equal to 60 micrograms/m2/day rhGM-CSF recovered neutrophil and platelet counts more rapidly, had fewer days with fever, and were discharged from the hospital sooner. PMID- 3042051 TI - Gene products which play a role in cancer invasion and metastasis. AB - Invasion requires a number of distinct tumor cell interactions with host tissue, beginning with attachment to the matrix, followed by hydrolysis of matrix material and locomotion. Gene products which may be involved in these steps are discussed here. Laminin receptors and integrins have roles in the adhesion phase, while certain collagenases are prominent among the matrix-degrading enzymes. Autocrine motility factors, distinct from growth factors, appear to be involved in tumor cell locomotion. Finally, certain oncogenes, particularly of the ras family, are closely related with metastatic potential. A detailed understanding of the molecular biology of invasion and metastasis could ultimately lead to specific means of interfering with or even reversing these malignant processes. PMID- 3042052 TI - Effects of human breast fibroblasts on growth and 17 beta-estradiol dehydrogenase activity of MCF-7 cells in culture. AB - The effect of conditioned medium from human breast fibroblasts on growth and 17 beta-estradiol dehydrogenase (E2DH) activity of the MCF-7 human breast cancer cell line has been investigated. Fibroblasts were derived from normal and tumorous (benign and malignant) breast tissue. Conditioned medium from normal derived fibroblasts inhibited the growth of MCF-7 cells, the effect being statistically significant by day 3 of culture. In contrast, conditioned medium from benign and malignant derived fibroblasts significantly enhanced the growth of MCF-7 cells by day 9 of culture. When added to MCF-7 cells for three days, conditioned medium from all three types of fibroblasts increased E2DH activity in the reductive direction (estrone (E1)----estradiol (E2] 2-3 fold. There was little or no effect on the oxidative direction (E2----E1). After 12 days, enzyme activity in the reductive direction was still markedly increased, the effect being greatest in conditioned medium from fibroblasts derived from malignant breast tissue, and least in conditioned medium from fibroblasts derived from benign breast tumors. These results demonstrate that human breast fibroblasts may have paracrine influences on neighboring epithelial cells in vivo. PMID- 3042053 TI - Adjuvant trial for stage II receptor-positive breast cancer: CMF vs. CMF + tamoxifen in a single centre. AB - The purpose of a randomized trial achieved in a single centre (Fondation Bergonie, Bordeaux, France) was to compare chemotherapy alone (intravenous CMF) versus chemotherapy and hormonotherapy (CMF plus tamoxifen-30 mg per day during 2 years), for patients with stage II breast carcinoma and positive values of estrogen and/or progesterone receptor (EPR) (greater than 10 and greater than 15 fmoles mg protein-1 respectively). Three hundred and thirty four women treated by surgery +/- radiotherapy are included in this trial from 06.01.81 to 12.31.84. No patient is lost for follow-up. Eight are excluded. Three hundred and twenty six patients are evaluable with a 38 month median follow-up. For EPR assay, the dextran charcoal micromethod was used in the same centre. The two groups are identical as far as age, hormonal status, TNM, EPR values, and histological features are concerned. Analysis of results shows a significant improvement of relapse free survival (p = 0.018) and also overall survival (p = 0.04) for the CMF+ tamoxifen group. PMID- 3042055 TI - Human megakaryocytopoiesis: in vitro regulation and characterization of megakaryocytic precursor cells by differentiation markers. AB - Understanding of human megakaryopoiesis has been improved by in vitro culture techniques, characterization of platelet proteins as differentiation markers and megakaryocyte purification. A continuum of megakaryocytic cells ranging from the CFU-MK (cell capable of proliferation) to the platelet has been demonstrated. CFU MK is a heterogeneous population of cells which do not express platelet proteins other than platelet GP IIb and IIIa which may be present in low concentration. The main platelet proteins are synthesized later during differentiation of a 2N cell, just before the polyploidization process. Several homogeneous growth factors GM-CSF, interleukin 3 and erythropoietin are able to sustain human megakaryocyte colony formation. However, no specific MK-CSF has yet been purified. Purification of factors active on late stages of differentiation has proved difficult when using in vivo techniques to test their activity. However, recently such a factor has been purified to homogeneity by testing its action on the biosynthesis of PF4 by megakaryocytes. A negative regulation of megakaryopoiesis has been suggested by the inhibition of MK-colony formation by platelet products especially TGF-beta. A better understanding of human megakaryopoiesis may be of importance as it may allow modifications of platelet production in a clinical setting. PMID- 3042056 TI - Heparin-induced thrombocytopenia. AB - Thrombocytopenia is a common adverse effect of heparin therapy which may occur with bovine or porcine heparin when it is given either intravenously or subcutaneously. There are two main clinical types: a common, mild thrombocytopenia of early onset, probably due to the platelet proaggregating effect of heparin itself and a severe delayed-onset thrombocytopenia caused by an immune mechanism. Patients with mild thrombocytopenia due to heparin are usually asymptomatic. However, patients with severe thrombocytopenia may develop thromboembolic complications which often result in disastrous sequelae such as limb gangrene necessitating amputation or death. The thromboembolic complications may be attributed to an IgG heparin-dependent platelet antibody which induces thromboxane production and platelet aggregation. The diagnosis of heparin-induced thrombocytopenia is made clinically and may be confirmed by the demonstration of the heparin-dependent antibody in vitro by platelet aggregometry or [14C] serotonin release. In patients with delayed-onset severe thrombocytopenia, heparin should be stopped and warfarin commenced. Antiplatelet drugs and/or dextran may also be added. Recently low molecular weight heparin has been used with some success. Conversely, heparin may be continued in patients with asymptomatic mild thrombocytopenia provided the patients' condition and the platelet counts are closely monitored. PMID- 3042058 TI - Management of patients with shock and sepsis. AB - The pathophysiology of shock associated with sepsis in complex, but granulocyte activation and production of toxic oxygen radicals, is of major importance in producing endothelial cell injury. Multiple organ failure including a reversible cardiomyopathy with impairment of left ventricular ejection fraction, are known factors complicating, and in the latter, perpetuating, shock. When treating the severely shocked patient, a systolic pressure of 100 mmHg and a PaO2 of greater than 8 kPa (60 mmHg) should be aimed for. Non-response to oxygenation, respiratory support, volume, and metabolic control, may be an indication for insertion of a thermodilution catheter into the right heart, so that cardiac output can be measured and oxygen delivery maintained above 10 mmol/kg/min. Where, by manipulation of respiratory indices and inotropic support, achievement of this level is not possible, the prognosis is grave. Antibiotic therapy is discussed, therapy being instituted if at all possible once the haemodynamic state has been improved. PMID- 3042057 TI - Myeloid cell lines: tools for studying differentiation of normal and abnormal hematopoietic cells. AB - Acute myeloid leukemia is caused by one or several transforming events which usually result in a block of myeloid precursor cell maturation. Human cell lines can serve as model for hematopoietic cells arrested at different stages of myeloid differentiation. These homogeneous populations help to dissect the cellular and molecular events involved in leukemogenesis, such as proto-oncogene activation. Furthermore, the efficacy and mechanism of action of compounds inducing differentiation of leukemic cells can be studied. Finally, these lines can permit the analysis of proliferation and differentiation of normal myeloid precursor cells. PMID- 3042054 TI - Progress in understanding breast cancer: epidemiological and biological interactions. AB - Little progress has been made recently in our understanding of the epidemiology of breast cancer. While results from epidemiologic studies regarding reproductive factors remain fairly reproducible from one study to another, other associations such as that between breast cancer risk and dietary fat intake, although biologically plausible, are not consistently found in direct study of humans, while yet other associations, which appear less plausible biologically, become stronger (such as the increased risk associated with modest levels of alcohol consumption). In this paper we attempt to review the epidemiology and biology of breast cancer jointly and describe possible mechanisms of breast cancer induction, the cellular composition of the breast, the epidemiology of breast cancer, and salient biological features, and attempt to reconcile the biology and epidemiology. It becomes obvious that future progress depends on better biological thinking by epidemiologists, and vice-versa. Areas of further research are suggested and discussed, concluding that the ability to measure diet with greater precision could have an important role to play in clarifying our understanding of breast cancer. PMID- 3042059 TI - Management of the patient with large cell lymphoma. AB - Lymphomas are neoplasms of B-lymphocytic (75% of cases) or T-lymphocytic (20% of cases) origin. It is now appreciated that the various sub-types of disease represent counterparts of the normal pathways of lymphoid differentiation and transformation, from resting cells to fully transformed and activated lymphoid cells. The large cell lymphomas represent a group of neoplasms, formerly grouped together and termed 'histiocytic lymphoma' which represent transformed lymphocytes, of T- or B-cell derivation. Within the Working Formulation for Clinical Usage, a recently proposed translational tool, these lymphomas fall within the 'Intermediate' or 'High-grade' disease categories. The initial evaluation of a patient with large-cell lymphoma must include a formal staging procedure, in order to ascertain the precise extent of disease, so that appropriate therapeutic decisions may be made. This evaluation is also useful for defining all sites of lymphomatous disease, so that the results of any given therapeutic intervention can be accurately assessed. For the patient with localized, stage I disease, one option would be to take the patient to formal staging laparotomy, in order to attempt to prove the presence of pathologic Stage I disease. In this instance, wide-field radiotherapy may be employed, with the chance of achieving long-term, disease-free survival of approximately 70%. Alternatively, the patient may be staged less aggressively, and given combination chemotherapy, which is also effective in low-stage disease. In patients with Stage II, III or IV large-cell lymphoma, multi-agent chemotherapy is the expected treatment modality, although some controversy currently exists regarding the optimal regimen(s).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3042060 TI - Molecular diagnosis of haematological neoplasms. AB - DNA analysis has become of practical value in the diagnosis and classification of leukaemias and lymphomas. This is exemplified by the study of lymphoproliferative disorders using immunoglobulin and T-cell receptor gene probes for the determination of clonality and cell lineage. Chromosomal analysis with DNA probes is now a useful complementary approach to cytogenetics. For example, the study of particular lymphomas or chronic myelogenous leukaemia with DNA probes hybridising to specific chromosomal breakpoints allows the detection of chromosomal translocations at a genomic level. Chromosomal loss in neoplastic cells can be detected by DNA probes in individual heterozygous for particular restriction fragment length polymorphisms, most efficiently by locus-specific hypervariable region probles. These techniques will enable progress to be made in the understanding of the biology of remission and disease progression in haematological malignancies. PMID- 3042061 TI - Vascular endothelium, haemostasis and thrombosis. AB - The vascular endothelium consists of a monolayer of cells. Integrity of the endothelium is essential for maintenance of blood fluidity as the subendothelium is composed of structures which rapidly activate platelets and coagulation. Recent research indicates that the endothelium is involved in other processes particularly vasoactivity, immune reactions and inflammatory processes. Much of the information has come from experiments using cells in culture and there has been considerable progress in defining synthesis and release of endothelial products involved. Of particular interest is the evidence for endothelial coagulant activity and exposure of binding sites for coagulation proteins, with the simultaneous suppression of antithrombotic activities, all of which can be brought about by inflammatory mediators. Despite the rapid progress in cell biology, clinical applications have tended to lag behind and there are still few aspects of endothelial cell metabolism and pathophysiology which can be precisely defined in patients with disorders in which the endothelium is without doubt actively involved. PMID- 3042062 TI - Long term warfarin treatment in artery disease. AB - The long-term use of oral anticoagulants like warfarin in artery disease has long been controversial. Possible aims of treatment include the primary or secondary prevention of systemic embolism, preventing recurrence after myocardial infarction or the progression of transient cerebral ischemia to a complete stroke, and the prevention of artery graft occlusion. The value of long-term anticoagulation is generally accepted in the few situations where, as in patients with mechanical heart valve prostheses, mitral valve disease and atrial fibrillation, or idiopathic dilated cardiomyopathy, the risk of arterial thromboembolism without anticoagulation is known to be high and there is good evidence that anticoagulants are effective, so the benefit:risk balance strongly favours their use. In many settings, however, it is hard to justify long-term warfarin treatment as the benefit:risk balance remains unknown; either because the risk of thromboembolism without anticoagulation remains to be clearly defined (as in the case of patients with 'lone' atrial fibrillation), or because possible benefits have not been well documented (as after transient cerebral ischemia or peripheral vascular surgery). Finally, there is the difficult problem of estimating the benefit from long-term anticoagulation after myocardial infarction. It seems that warfarin can reduce the likelihood of non-fatal reinfarction with relative safety in highly selected patients, but whether it reduces mortality, and how its effect compares with that of other, more recent, therapies aimed at preventing reinfarction, remains unknown. PMID- 3042063 TI - Immunocytochemical localisation of GABA in endocrine cells of the rat entero pancreatic system. AB - The occurrence of GABA-containing cells in the rat entero-pancreatic system was investigated by using anti-GABA-glutaraldehyde antibodies at the light and electron microscope level. In the pancreas, the B cells showed intense immunoreactivity, contrary to non-B and exocrine cells. Moreover, post-embedding immunogold staining was localised mostly in mitochondria, close to rough endoplasmic reticulum and in the nucleus. The insulin granules appeared nonsignificantly stained, which suggests the lack of cosecretion of GABA together with insulin. In the duodenum, GABA immunoreactivity was detected in certain endocrine cell types, suggesting a possible interaction with this amino acid. The well established GABAergic innervation in the enteric system was also confirmed by immunolabelling. PMID- 3042064 TI - Proceedings of the XIVth annual meeting of the European Cooperative Group for Bone Marrow Transplantation. 10-13 April 1988, Chamonix, France. Abstracts. PMID- 3042065 TI - Borderline pathology in post-incest female adolescents. Diagnostic and theoretical considerations. PMID- 3042066 TI - The psychology of combining dynamic psychotherapy and alcoholics anonymous. PMID- 3042067 TI - Pathogenesis of ankylosing spondylitis and rheumatoid arthritis. Proceedings of the second international symposium. 14-15 April 1987, London. Dedicated to Dr. D. C. O. James. PMID- 3042068 TI - Pathogenesis of ankylosing spondylitis: the state of the art. PMID- 3042069 TI - Historical aspects of the aetiology of rheumatoid arthritis. AB - Since the first description of the disease in the early part of the nineteenth century, the aetiology of rheumatoid arthritis has been associated with the interacting concepts of diathesis or internal factors and environment or external factors. PMID- 3042070 TI - Pathophysiology of chronic synovitis. PMID- 3042071 TI - Molecular biology of the HLA-B27 locus. AB - The molecular biology of the HLA-B27 locus is reviewed. The HLA-B27 gene itself does not differ between healthy individuals and ankylosing spondylitis patients. Several unique features of the HLA-B27 molecule have been identified and one epitope was proposed to cross-react with bacterial proteins. PMID- 3042073 TI - Pathogenetic factors in rheumatoid synovitis. PMID- 3042074 TI - The Middlesex Hospital prospective study of early rheumatoid disease. AB - A review of findings obtained from a follow-up study of rheumatoid arthritis implicated a positive rheumatoid factor test as the most useful indicator of subsequent joint erosion. PMID- 3042072 TI - Induced expression of class II transplantation antigens in the cartilage-pannus junction in RA: chronic synovitis as a model system for aberrant T-lymphocyte activation. AB - An induced expression of class II transplantation antigens is seen on synovial cells in human arthritides such as rheumatoid arthritis (RA) and ankylosing spondylitis (AS) as well as in some experimental animal models for arthritis, i.e. in collagen arthritis in mice and rats and in adjuvant arthritis in rats. In this paper, additional original data are presented concerning the expression of HLA-DR, DP, and DQ antigens on macrophage-like and fibroblast-like cells within the cartilage-pannus junction in RA. The functional implications of this induced synovial class II antigen expression is discussed against the background of both in vitro experiments on human synovial cells and in vivo experiments in the animal arthritis models. PMID- 3042075 TI - Lymphocyte response to enterobacterial biostructures in seronegative spondarthritis: specific T-cell mediated immunity or non-specific polyclonal B cell activation? AB - The proliferative response, as measured by thymidine uptake and the B-cell differentiation response, as measured by Ig secretion, have been studied in peripheral blood mononuclear cells obtained from patients with ankylosing spondylitis, Yersinia reactive arthritis, Klebsiella infections and healthy controls, following stimulation with enterobacterial biostructures. PMID- 3042076 TI - Molecular mimicry: fact or fiction? AB - Molecular studies suggest that peptide sequence similarities are present between HLA-B27 and bacteria, but the pathological mechanisms in spondyloarthropathies remain unclear. PMID- 3042077 TI - Klebsiella antibodies in ankylosing spondylitis and Proteus antibodies in rheumatoid arthritis. AB - Antibodies to Klebsiella, but not to other bacteria, have been shown to be present in patients with active ankylosing spondylitis (AS) by seven different techniques. Antibodies to Proteus, but not to E. coli or Klebsiella, have been shown to be present in patients with active rheumatoid arthritis (RA) by three different techniques. It is suggested AS and RA are forms of reactive arthritis, to Klebsiella and Proteus respectively, probably mediated by cross-reactivity to HLA antigens. PMID- 3042078 TI - Antibodies to Klebsiella and Proteus microorganisms in ankylosing spondylitis and rheumatoid arthritis patients measured by ELISA. AB - Antibodies to Klebsiella oxytoca and Proteus mirabilis in 21 active ankylosing spondylitis (AS), 13 active rheumatoid arthritis (RA), 19 inactive RA patients and 18 healthy controls were measured using enzyme-linked immunosorbent assay (ELISA). Elevated anti-Klebsiella antibodies were demonstrated in active AS patients compared to active RA (p less than 0.01), inactive RA patients (p less than 0.001) and controls (p less than 0.005). When measuring antibodies to Proteus, active RA patients showed higher levels of antibodies compared to active AS patients (p less than 0.005) and healthy controls (p less than 0.05). Further investigations are required to assess the role of Klebsiella and Proteus microorganisms in AS and RA respectively. PMID- 3042079 TI - Antibodies to Proteus in rheumatoid arthritis. AB - Increased levels of Proteus antibodies were found in patients with rheumatoid arthritis and coeliac disease, when compared to normal controls or patients with SLE and sarcoidosis. PMID- 3042080 TI - Ileocolonoscopic findings in seronegative spondylarthropathies. AB - Ileocolonoscopy with biopsy of caecum, ileocaecal valve and terminal ileum were performed on 232 patients with seronegative spondylarthropathies and on 65 control patients. Inflammatory gut lesions were found in 65% of the patients with reactive arthritis (ReA) and in 57% of the patients with ankylosing spondylitis (AS), especially in those with peripheral arthritis. The controls had a normal gut. This finding would suggest that exogenous factors causing inflammation of the gut lead to a disturbed permeability of the gut wall or to a deficient local immunological defence mechanism permitting antigens to enter the circulation, inducing the joint and tendon inflammation. Support for this hypothesis was provided by the results of a repeat ileocolonoscopy, disclosing a strong association between the presence of gut inflammation on biopsy and the persistence of joint inflammation. Patients presenting some of the histological lesions found on biopsy (especially active chronic lesions) and patients with proven Crohn's disease were found to share a genetic marker (HLA-BW62). This would suggest that some of the patients with seronegative spondylarthropathies suffer from a subclinical form of Crohn's disease of which the joint symptoms are the unique clinical manifestation. PMID- 3042081 TI - Urolithiasis in Jordanian children. A report of 52 cases. AB - Fifty-two children with urinary calculi seen between 1975 and 1986 were reviewed. Males dominated the series. The age distribution ranged from 10 months to 14 years (mean 7.2 years); 71% presented after school age. Most patients had upper tract stones. The main presenting symptoms were abdominal pain, infection and haematuria. The causative factors or co-factors were infection, malformations and urodynamic abnormalities. Metabolic disorders were rare. Calcium oxalate and uric acid stones were found most often. Surgical management was required in 88% of patients and only 3.8% had a recurrence. Presenting symptoms are variable and so a high index of suspicion is required for diagnosis. PMID- 3042082 TI - Combined free autologous bladder mucosa/skin tube for urethral reconstruction: an update. AB - A combined free autologous bladder mucosa/skin graft was used to reconstruct the urethra in 23 patients. Although the complication rate remains high (61%), the results represent a significant improvement over the use of complete mucosal tubes to the tip of the penis. The technique is recommended for those patients with failed hypospadias surgery, where sufficient skin is not available for urethral reconstruction. PMID- 3042084 TI - Caliceal-peritoneal fistula complicating percutaneous biopsy of a transplant kidney. PMID- 3042083 TI - Prostacyclin in the treatment of painful Peyronie's disease. AB - The effect of a prostacyclin infusion was studied in 5 men with persistent pain associated with Peyronie's disease. Marked side effects were associated with the infusion, which was of limited benefit. PMID- 3042085 TI - Immunosuppression in animals. PMID- 3042086 TI - Lipid storage myopathy associated with low acyl-CoA dehydrogenase activities. AB - A man with a painful proximal myopathy had excess lipid deposition in skeletal muscle, excretion of dicarboxylic acids in urine and low acyl-CoA dehydrogenase activities in skeletal muscle mitochondria. In addition he had little immunoreactive short-chain and medium-chain acyl-CoA dehydrogenase enzyme protein compared with normal controls. Following treatment with riboflavin there was considerable improvement in his clinical condition which was confirmed by further biochemical and morphological investigations. PMID- 3042087 TI - Widespread distribution of the c-src gene product in nerve cells and axon terminals in the adult rat brain. AB - The regional and cellular distribution of the proto-oncogene product pp60c-src, a member of the family of membrane-associated tyrosine-specific protein kinases, was analysed in adult rat brain. High-resolution SDS-PAGE allowed analysis of both the 'fibroblast' 60-kDa form and a variant, 61-kDa neuron-specific form of the c-src gene product which is encoded by an alternately processed c-src mRNA. Studies of microdissected brain regions showed that all CNS regions contained both forms of the enzyme, the 61-kDa form predominating in most regions with high content of gray matter and high density of synapses. Lesion-induced degenerations of specific neuronal elements in the basal ganglia decreased the level of both forms of the c-src gene product both in regions where cell bodies had been lesioned and in regions where nerve terminals had degenerated. The 61-kDa form of the enzyme appeared somewhat more sensitive to the effects caused by these lesions than the 60-kDa form. These results indicate that, within the mature mammalian brain, both cell body regions and nerve terminals of many, and possibly all, nerve cells contain both forms of the c-src gene product, the 61-kDa form being most highly enriched in the nerve cells. These results suggest that the enzyme may be involved in pleiotropic functions, including signal transduction in nerve terminals. PMID- 3042088 TI - An immunocytochemical and biochemical study of the microtubule-associated protein MAP-2 during post-lesion dendritic remodeling in the central nervous system of adult rats. AB - A monoclonal antibody against the microtubule-associated protein MAP-2 was used to examine the fate of this molecule during post-lesion dendritic remodeling in the hippocampus and septum of adult rats. Qualitative and quantitative immunocytochemical analyses were carried out in the dentate gyrus after unilateral destruction of the entorhinal cortex (EC). An increase in MAP-2 immunoreactivity was detected in dendritic processes located in the outer 2/3 of the ipsilateral molecular layer (ML) 2 days after the lesion. whereas dendritic staining decreased considerably in the inner 1/3 of the same ML. The increase of staining was also detected 4, 6 and 8 days after the lesion; it was accompanied by an increase in the immunoreactivity in the inner 1/3 of the ML. After that period, a progressive decrease in anti-MAP-2 staining toward control levels was detected along the whole extent of the ipsilateral ML. This was concurrent with alterations in dendritic orientation, and a decrease in stained dendrites in the inner 1/3 of the ML. By 30 days post-lesion anti-MAP-2 staining was almost identical to that of the contralateral ML, although the alterations in dendritic morphology were still present in the ipsilateral ML. Changes in MAP-2 levels were also evaluated by densitometry of Western blots or dot immunobinding of hippocampal extracts obtained at different post-lesion intervals. The results obtained revealed a pattern of change in MAP-2 levels identical to that observed with the immunohistochemical stain. A similar, immunocytochemical and biochemical, analysis conducted in the lateral septal nucleus after unilateral transection of the fimbria showed no changes in the distribution and/or content of MAP-2 at any post-lesion interval analyzed (2, 10 and 20 days post-lesion). The present observations show that post-lesion dendritic remodeling is concurrent with modifications in the levels and distribution of MAP-2. These modifications suggest that the dendritic cytoskeleton is dynamically changing in response to perturbation of the synaptic environment. In addition, our results indicate that these changes may only occur in those neurons which have the capability to remodel their post-synaptic surface in response to deafferentation. PMID- 3042089 TI - Identification in rodents and other species of an mRNA homologous to the human beta-amyloid precursor. AB - The isolation and sequencing of the core peptide (beta-amyloid) found in the plaques of patients with Alzheimer's disease has allowed the identification of a cDNA for the precursor protein. Using a human cDNA clone for this beta-amyloid material, we have identified an homologous mRNA (3.8 kb) in brain tissue obtained from 8 additional species. We have also determined its distribution in 7 brain regions and 12 organs obtained from rodents. A prominent, second mRNA species (2.2 kb) has been identified in rat non-neuronal tissues. The beta-amyloid gene is amply expressed in the brain of all vertebrates tested and in most rodent organs, indicating that it encodes a highly conserved and ubiquitous protein. PMID- 3042090 TI - Response properties of fibers in the hamster superior laryngeal nerve. AB - The purpose of the present investigation was to record electrophysiological responses from single fibers in the hamster superior laryngeal nerve (SLN) that were responsive to chemical stimulation of the larynx. Twenty chemical solutions, commonly used in studies of mammalian gustatory physiology, were applied to taste buds on and around the epiglottis. These stimuli were dissolved in physiological saline. Responses were the number of impulses elicited over a 15-s period following stimulus onset, above or below the background activity elicited by a previous rinse with saline. Unlike fibers in the hamster chorda tympani or glossopharyngeal nerves, SLN units were not easily classifiable into response types. Excitatory stimuli were primarily acids and bitter-tasting stimuli, with the order of their effectiveness being urea much greater than tartaric acid greater than HCl greater than KCl greater than citric acid greater than caffeine greater than quinine hydrochloride greater than acetic acid. The sweet-tasting stimuli and most salts other than KCl were primarily inhibitory, with the order of inhibitory effectiveness being CaCl2 greater than sucrose greater than fructose greater than LiCl greater than NaNO3 greater than Li2SO4 greater than NaCl. A hierarchical cluster analysis of fibers yielded no distinct clusters, yet differing sensitivities across the fibers were suggested. SLN fibers are highly responsive to sour and bitter stimuli, although they are not sensitive to fine differences in taste quality, as are fibers in other gustatory nerves. PMID- 3042091 TI - Apparent involvement of protein kinase C in the central glucoregulatory action of insulin. AB - We have studied the possible involvement of the calcium- and phospholipid/diacylglycerol-dependent enzyme, protein kinase C (PKC) in mediating insulin action in the central nervous system (CNS) by testing the effect of direct activation or blockade of the CNS PKC system on the plasma glucose responses to central insulin injection in mice. Insulin (0.1-1 microgram), injected into the CNS, produced rapid transient hypoglycemia. This effect appeared to involve interaction of insulin with specific receptors, since insulin analogs exhibiting diminished receptor binding affinity and peripheral bioactivity compared to the native hormone were much less active (i.e., insulin much greater than acetyl 3 insulin greater than proinsulin greater than IGF-I) or not active at all (i.e., insulin chain A and chain B). Central injection of the specific PKC activator, 12-O-tetradecanoylphorbol-13-acetate (TPA) (0.01-0.5 microgram), but not the inactive TPA analog, 4-alpha-phorbol or the unstable synthetic diacylglycerol analog, 1-oleoyl-2-acetyl-sn-glycerol (OAG), significantly enhanced the hypoglycemic response to co-administered insulin (0.5 microgram) or the insulin derivative, acetyl 3 insulin (2.5 micrograms). Central TPA had no effect on basal glucose levels. Furthermore, central administration of the selective PKC blockers, polymyxin B (PMB, 1-25 micrograms) or 1-beta galactosylsphingosine (psychosine, 0.5-10 micrograms) but not their respective inactive analogs, polymyxin E and sphingomyelin, strongly inhibited the hypoglycemic response to insulin (1 microgram) or acetyl 3 insulin (5 micrograms). PMB and psychosine, injected alone had no effect on basal glucose levels.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3042092 TI - Cerebroventricular dilation in spontaneously hypertensive rats (SHRs) is not attenuated by reduction of blood pressure. AB - In previous studies, we found that spontaneously hypertensive rats (Okamoto-Aoki SHRs) suffer progressive postnatal dilation of the brain ventricles. In the present study we examined intracerebroventricular pressure and blood pressure as possible mechanisms of ventricular dilation in SHRs. We found that intracerebroventricular pressure was not elevated in SHRs. The role of blood pressure was examined in SHRs treated chronically with the antihypertensive drug, captopril, beginning in utero, and in renal hypertensive Sprague-Dawley rats (SDs). Although our experimental treatments produced significant changes in mean arterial pressures, they did not alter brain ventricular size: SDs with experimental hypertension had normal-sized brain ventricles and SHRs with pharmacologically reduced blood pressure had enlarged ventricles. These results suggest that neither increased intraventricular pressure nor high blood pressure is the sole cause of hydrocephalus in SHRs. PMID- 3042093 TI - Ultrastructural evidence suggests variations in biosynthesis and processing within LH-RH neurons as a function of ovariectomy in rats. AB - We have demonstrated that populations of luteinizing hormone-releasing hormone (LH-RH)-immunopositive neuronal perikarya change following gonadectomy of male and female rats in a sex-dependent manner related to rises in plasma luteinizing hormone (LH). In this study we characterize the ultrastructural state of organelles involved in protein synthesis, primarily within perikarya of rostral preoptic area LH-RH neurons surrounding the organum vasculosum of the lamina terminalis (OVLT). One day following ovariectomy little rough endoplasmic reticulum (RER) was evident, however, the cisternae were heavily laden with ribosomes and numerous polysomes were present free in the cytoplasm. Six days and 3 weeks post-ovariectomy the cisternae of RER were progressively more abundant and dilated; the multiple Golgi apparati, located in close proximity to the RER, were composed of many lamelae and extensive associated vesicles. We propose that increased pools of messenger ribonucleic acid (mRNA) are generated by 1 day post ovariectomy prior to increased synthesis of precursor, 6 days and 3 weeks post ovariectomy. Axodendritic synaptic profiles in the neuropil surrounding LH-RH perikarya increased in number in the rostral medial preoptic area and lateral anterior hypothalamic area. We conclude that removal of gonadal steroids results in greater biosynthetic activity in LH-RH neurons, and suggest that the enhanced biosynthesis is related to increases in afferent activity. PMID- 3042094 TI - Inhibition of stress-induced hyperglycemia by tail pinching or intraventricular enkephalin administration in the rat. AB - The tail pinch (t-p) method added to a basal restraint stress produced inhibition of the stress-induced hyperglycemia, an effect that was neutralized with intrathecal anesthesia but not with intracerebroventricular (i.c.v.) naloxone (50, 100, 1000 ng/100 g) or with intraperitoneal naloxone injections (0.1-0.3 mg/100 g). A similar negative result was obtained with i.c.v. administration of 500 and 1000 ng/100 g of beta-endorphin. In contrast, a single i.c.v. injection of 1000 ng/100 g of Met-enkephalin reproduced the t-p inhibitory effect. The latter was not elicited by i.c.v. FK 33824, an enkephalin analogue, a result that supports the specific participation of the delta-opioid receptors. The results obtained with central alpha-adrenoceptor antagonists and central noradrenergic chemical destruction, or central alpha-adrenoceptor agonists, support the production of a reinforcement of the alpha-adrenoceptor stress stimulation by the t-p procedure, probably through noradrenaline and enkephalin mediation. PMID- 3042096 TI - Vasoactive intestinal polypeptide (VIP)-like immunoreactivity in the suprachiasmatic nucleus of the perinatal rat. AB - Vasoactive intestinal polypeptide (VIP)-like immunoreactivity was examined in the suprachiasmatic nucleus (SCN) of the perinatal rat using the unlabeled antibody enzyme method of Sternberger. Our results showed that VIP-like immunoreactivity was present in cells and fibers of the SCN prior to birth. Immunoreactive neurons at this time appeared morphologically immature, and their distribution differed somewhat from that seen in the adult SCN. PMID- 3042095 TI - Light-microscopic immunolocalization of the growth- and plasticity-associated protein GAP-43 in the developing rat brain. AB - Growth-associated protein-43 (GAP-43) is a developmentally regulated, fast axonally transported phosphoprotein whose synthesis and transport are enhanced during periods of growth and synaptic terminal formation. GAP-43 is a substrate of protein kinase C and is identical to protein F1, a phosphoprotein which is regulated during long-term potentiation in the hippocampus. In order to characterize the cellular localization of GAP-43, we have raised a specific antiserum against it, and used this as a probe to show that GAP-43 is neuron specific, and is localized to growing neuronal processes in developing rat brain, and to presynaptic terminals in both the peripheral and central nervous system. In the mature CNS, GAP-43 immunoreactivity is present in most neuropil areas, but is especially dense in the molecular layers of the cerebellum, neocortex, and the hippocampus, structures known to exhibit synaptic plasticity. Its localization, together with biochemical data concerning the dynamics of its synthesis and its identity as protein F1, suggest that GAP-43 may be involved in axon growth in the developing nervous system, and in some aspect of synaptic plasticity in the mature CNS. These data also suggest that axon growth and synaptic plasticity in the brain may be regulated by a common mechanism, both involving the protein kinase C-mediated phosphorylation of GAP-43. PMID- 3042098 TI - [Has immunotherapy a place in insulin-dependent diabetes of children?]. PMID- 3042097 TI - Estrogen and insulin synergism in neurite growth enhancement in vitro: mediation of steroid effects by interactions with growth factors? AB - Addition of estradiol to organotypic cultures of the fetal murine hypothalamus, preoptic area and cerebral cortex has been shown to elicit a striking enhancement of neurite growth which appears restricted to estrogen receptor-containing explant regions. The mechanisms underlying this response are unknown. An important question is whether the neurite enhancement which follows exposure to estradiol is due directly to the interaction of estrogen with the cell that was stimulated (the receptor-containing cell) or whether intermediate steps involving the possible interaction of estrogen and the endogenous polypeptide neurite promoting growth factors or their receptors may play an important role. Recent findings in the cultures suggest that the effect of estrogen on neurite growth may involve synergistic interactions between estradiol and insulin-related peptides and may be important in regulating estrogen-responsive neurite growth in the central nervous system. Concurrent addition of estradiol and high levels of insulin (10 micrograms/ml or 50 micrograms/ml) to cultures of the olfactory bulb, hypothalamus, preoptic area and cerebral cortex of the fetal rat and mouse results in a dramatic acceleration and increase of neurite outgrowth which appears localized to estrogen receptor-containing explant regions. The supraphysiological concentrations of insulin required to elicit this response suggest that the factor(s) involved is unlikely to be insulin per se. Insulin may activate the receptor of different but closely related molecules such as the insulin-like growth factors (IGF)-I or -II to which it exhibits a relatively low affinity. Interactions between hormones and endogenous growth factors have been implicated in the modulation or mediation of an increasing number of endocrine dependent, differentiative processes in vivo and in vitro.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3042100 TI - [Epidemiology and prevention of keratitis in rice harvesters]. PMID- 3042099 TI - [Sympathetic renal nerves and genetic hypertension in the rat]. PMID- 3042101 TI - [Use of deltamethrin for the insecticidal treatment of cereal and leguminous seeds after harvesting and of green coffee beans]. PMID- 3042102 TI - [Use of pirimiphos-methyl for the treatment of cereals after harvest]. PMID- 3042103 TI - [Viennese psychiatry between Freud and Wagner von Jauregg]. PMID- 3042104 TI - [Impact of weak neonatal estrogenization of the male rat on his adult sexual behavior. Medical considerations]. PMID- 3042105 TI - [Arterial hypertension and diabetes mellitus: value of converting enzyme inhibitors]. PMID- 3042107 TI - [Problems of mothers and children in French-speaking countries of Africa]. PMID- 3042108 TI - [The hospital in French-speaking tropical Africa]. PMID- 3042106 TI - [Universal treatment of ophthalmic migraine]. PMID- 3042109 TI - [Eulogy of Marcel David (1898-1986)]. PMID- 3042110 TI - [The decline of the regional medical press]. PMID- 3042111 TI - [Painless myocardial ischemia: what prognosis? what attitude?]. PMID- 3042112 TI - [Epidemiology and vital prognosis of perforated ulcers of the duodenal bulb]. PMID- 3042113 TI - The yeast aminopeptidase Y. AB - A metal-dependent aminopeptidase (EC 3.4.11.-), designated APase Y, has been purified to homogeneity by conventional methods. The enzyme is composed of a single polypeptide chain with molecular mass of 102 kilodaltons, estimated by sodium dodecyl sulphate - polyacrylamide gel electrophoresis, with a blocked N terminal amino acid. It possesses neither endopeptidase nor carboxypeptidase activity and is strongly inhibited by metal-chelating agents, Zn2+, and the protein inhibitor from Neurospora crassa. APase Y is insensitive to Cl anions, S- S reducing reagents, serine protease inhibitors, and the peptidase inhibitor benzamidine. Co2+, Hg2+, and p-chloromercuribenzoate can activate the enzyme up to 22, 20, and 55%, respectively. The holoenzyme is resistant to yeast endopeptidases A, B, and Y, whereas the apoenzyme (obtained after treatment with chelators) is susceptible to the serine endopeptidases B and Y. The enzyme catalyzes hydrolysis of most L peptides possessing free alpha-amino (or imino) group by stepwise removal of N-terminal residue. Peptides with L-leucine at the N terminus are cleaved preferentially. The enzyme is unable to catalyze hydrolysis of X--Pro type peptide bonds, and inefficiently hydrolyzes bonds between Asp--X and Glu--X. L-leucine p-nitroanilide hydrolyzes optimally at pH 8.2 with a Km value of 1 mM. The purified enzyme is stable during storage in 0.05 M phosphate buffer, pH 6.7, containing 40-50% glycerol, at -20 degrees C. PMID- 3042114 TI - Suppression by the ColV,I-K94 plasmid of inhibitor sensitivities in ompA mutants of Escherichia coli. AB - A series of ompA mutants derived from Escherichia coli K12 strains showed increased sensitivity (compared with the ompA+ parents) to aminoglycoside antibiotics and to other cationic agents including polymyxin B. One tested mutant also showed increased sensitivity to nafcillin and fusidic acid, but not to the hydrophilic ampicillin. All these inhibitor sensitivities in the ompA mutants were suppressed by ColV, I-K94 and by certain other ColV plasmids, but not by any of the other tested large plasmids. Suppression correlated with the production of the VmpA protein, but transfer and colicin components were not needed for suppression. Further comparison of the ompA and vmpA genes and their products was made and it indicated that there is little if any homology between the genes, that the synthesis of their products is regulated by quite different mechanisms, and that regions of these gene products exposed at the cell surface show different susceptibility to protease attack after denaturation. PMID- 3042116 TI - Transduction of Escherichia coli in soil. AB - Bacteriophage P1-mediated generalized transduction of Escherichia coli K-12 was assessed in nonsterile soil. Auxotrophic recipient cells (thr- leu- thi- rpsL) were incubated in a sandy and a silty clay loam soil, and the transducing phage lysates from prototrophic strains carrying transposon 10(Tn10) in either purE or aroL regions were added. At intervals, the bacterial populations derived from the soils were plated on selective-differential media to enumerate prototrophic (thr+, leu+, or Tcr) transductants. Of 100 bacterial isolates obtained on the selective-differential media, 58 (14 thr+; 11, leu+; 33 Tcr) were confirmed E. coli transductants. The frequency of transduction in soil was ca. 10(-6). These data demonstrate the potential use of bacteriophage P1 to genetically manipulate E. coli in situ. PMID- 3042115 TI - Physiological regulation of protease activity in Streptomyces peucetius. AB - Streptomyces peucetius ATCC 29050, a producer of anthracycline antineoplastic agents, was investigated for the expression of intracellular and extracellular azocaseinase activities as a function of growth and medium conditions. When cultures were grown in either nitrate-containing defined medium or glucose-yeast extract complex medium, the intracellular proteolytic activity was greatest during early to mid stationary phase, whereas the extracellular proteolytic activity was produced in late stationary phase. All of the proteolytic activity detected against azocasein was of a serine-type protease activity. These late occurring proteases may have some function in cellular turnover associated with secondary metabolism and (or) morphogenesis. PMID- 3042117 TI - An examination of HIV antibody testing. PMID- 3042119 TI - Norfloxacin: a new quinolone. Committee on Antimicrobial Agents, Canadian Infectious Disease Society. PMID- 3042120 TI - Low vision services: a model for sequential intervention and rehabilitation. PMID- 3042121 TI - Smoking prevention for Ontario school children: we know what works, now let's make it happen. PMID- 3042118 TI - [Hypothesis of the changes in the frequency of asthma]. AB - Review of the literature reveals sporadic but true increases in the prevalence and death rates for asthma over the past 25 years. Although changes in hereditary, allergic and environmental factors must be considered as possible causes of this increase, its suddenness points to risk factors that change rapidly. Such is the case with atmospheric pollutants, notably nitrogen dioxide and particles, both of which have been shown to have adverse effects on the tracheobronchial tree. These pollutants tend to concentrate inside homes, especially since the early 1980s, when the energy crisis brought about changes in the home environment. PMID- 3042122 TI - Sex differences in cigarette smoking in Canada, 1900-1978: a reconstructed cohort study. PMID- 3042124 TI - Bone marrow transplantation in children. Nursing management of late effects. PMID- 3042123 TI - Mortality attributable to tobacco use in Canada. PMID- 3042125 TI - Evaluations of death education in nursing. A critical review. PMID- 3042126 TI - Double-blind placebo-controlled trial with flunarizine in therapy-resistant epileptic patients. AB - The anticonvulsant efficacy and side-effect liability of flunarizine (15 mg/day) was investigated in a randomized, double-blind, placebo-controlled, crossover design in 30 outpatients with drug-resistant complex partial seizures. Flunarizine or placebo was added to the preexisting medication and each patient was followed up for 10 months. At the end of the study data from 22 patients were available for evaluation. In patients taking first flunarizine and then placebo, plasma levels of flunarizine were still detectable at the end of the 4 months' placebo phase. In the group of 13 patients starting therapy with placebo, a significant seizure frequency reduction was observed during the flunarizine period in 11 patients, whereas one patient showed no change and seizure frequency increased in another patient. Two patients had a 50% reduction in seizure frequency. Flunarizine was well tolerated and few side effects were noted. PMID- 3042127 TI - Loco-regional recurrent melanoma: II. Non-systemic treatments (1964-1979). PMID- 3042128 TI - Loco-regional primary and recurrent melanoma: III. Update of natural history and non-systemic treatments (1980-1987). PMID- 3042129 TI - Paraplatin (carboplatin): new advances in cancer therapy. Update on clinical experiences. Proceedings of a satellite symposium. Madrid, Spain, 2 November 1987. PMID- 3042130 TI - Simultaneous treatment with carboplatin (NSC-241-240) and radiotherapy in advanced unresectable squamous cell carcinomas of the head and neck. PMID- 3042132 TI - Neurosurgical complications of pediatric orthotopic liver transplantation. AB - Liver transplantation is the only definitive treatment of end-stage liver disease. The University of Nebraska began its hepatic transplantation program in July 1985. Since that time, 43 children and 48 adults have undergone orthotopic liver transplantation (OLTx) with survival rates to date of 79.1% and 79.2%, respectively. Eight children developed complications of neurosurgical interest (18.6% incidence). Hemorrhagic complications were the most frequent. Neurosurgical salvage was achieved in five patients, but delayed complications of the transplant caused the death of two of these children. Two survivors are functioning well at home and in kindergarten, one child is doing well but is still hospitalized, and one child is vegetative. Aggressive management of life threatening CNS problems is thus appropriate in this population. The authors review the pathophysiology of these complications, as well as potential pitfalls in their management. PMID- 3042133 TI - An attempt to prevent the problem of shunt-tube migration. AB - Five children are presented with complications related to migration of peritoneal shunt tubes. Anchoring the distal end of the peritoneal tube to the peritoneum was found to be a satisfactory answer to the problem in 13 children. This simple method does not add much to the operation time and has prevented shunt-tube migration in the group studied. PMID- 3042131 TI - Hydromyelia: a critical review. AB - The author reviews current concepts involving the etiology, pathogenesis, and treatment of hydromyelia. A critical evaluation of current surgical approaches in relation to the type of hydromyelia present is discussed. PMID- 3042134 TI - Ventricular shunt functioning despite extraventricular location of the catheter tip as revealed by computed tomography. AB - Two cases of infantile hydrocephalus are reported, in which the ventricular shunt proved to function despite the catheter tip being located outside the ventricular system, as revealed by computed tomography. This condition could be erroneously diagnosed as arrested hydrocephalus or proximal shunt malfunction, with a resultant potential risk of inadequately removing the catheter. PMID- 3042135 TI - Modified method for prophylaxis against unishunt system complications with presentation of total intraventricular migration of unisystem ventriculoperitoneal shunt. AB - This report is a case of shunted hydrocephalus in which the Unishunt system was placed without the clips (lock and slip). Upward migration of the whole system into the lateral ventricle was discovered 1 month after surgery. The shunt was reinserted, placing the proximal "lock-clip" under the cranial flap and the distal "slip-clip" subcutaneously at the abdominal area. The skin over the slip clip ulcerated, so it was repositioned and fixed deep in the abdominal muscle, extraperitoneally. Different factors caused the migration: negative sucking intraventricular pressure; pushing, positive intra-abdominal pressure; the course of the subcutaneous tract of the tube not being straight; incorrect fixation of the ends of the system; faulty handling of the baby. Choosing the type of shunt, its pressure opening, antisiphon device, uni- or multi-component, and following a straight line between the ventricular and abdominal inlets are all important factors that guard against migration and assure fixation. The modified fixation method using clips is recommended as a routine technique, especially in emaciated babies, to guard against skin ulceration. PMID- 3042137 TI - [Acute ventricular tachycardia]. PMID- 3042138 TI - [Preliminary analysis of an expert system for bloodless cardiologic diagnosis]. PMID- 3042139 TI - [Physiopathologic and clinical significance of 24-hour behavior of arterial pressure and cardiac frequency in recent heart transplants]. PMID- 3042140 TI - Special issue: Tribute to Julius Axelrod on the occasion of his 75th birthday. PMID- 3042136 TI - Spinal meningeal malformations in children (without meningoceles or meningomyeloceles). AB - Multiple meningeal malformations are described: anterior or lateral meningoceles, extradural meningeal cysts, and intradural arachnoid cysts. All diverticulae appear to be extensions of the subarachnoid space, producing symptoms early or later. It is impossible to unify all these lesions because they cause multiple pathological conditions, depending upon the anatomical form or level, other systemic malformations, spinal abnormalities, or associated familial diseases. Surgical treatment requires complete evaluation of each anatomical aspect before procedure. PMID- 3042141 TI - Molecular mechanisms of exocytosis: the adrenal chromaffin cell as a model system. AB - 1. The release of neurotransmitters, hormones, and enzymes involves exquisitely regulated events which ultimately result in the fusion of the secretory vesicle with the cell's plasma membrane, releasing the vesicle contents into the extracellular space. 2. The biochemical and cellular mechanisms mediating exocytosis have been extensively studied in a model system of primary cultured adrenal chromaffin cells. 3. This paper briefly reviews current understanding, and directions of future studies in exocytosis using this model system. PMID- 3042142 TI - Phenol sulfotransferase inheritance. AB - 1. Phenol sulfotransferase (PST) catalyzes the sulfate conjugation of many phenolic and catechol neurotransmitters. Human tissues contain both thermostable (TS) and thermolabile (TL) forms of PST that differ in their substrate specificities, inhibitor sensitivities, physical properties, and regulation. 2. Individual variations in the levels of activity of both TS and TL PST in the human platelet are strongly influenced by inheritance. 3. Individual differences in the level of platelet TS PST activity are correlated with individual variations in the activity of this form of the enzyme in human cerebral cortex, liver, and intestinal mucosa. 4. There are also individual familial differences in the thermal stability of TS PST in the platelet. These differences are correlated with individual variations in the thermal stability of TS PST in cerebral cortex, liver, and intestinal mucosa. 5. Individual variations in the thermal stability of TS PST in hepatic tissue are associated with the presence of one or both of a pair of TS PST isozymes that can be separated by ion-exchange chromatography and that differ in their thermal stabilities. 6. This series of observations suggests that a structural gene polymorphism may be one mechanism by which inheritance controls TS PST in humans. The isozymes of TS PST in liver may represent the products of alternative alleles for this polymorphism, alleles that might control the structure of TS PST in many human tissues. PMID- 3042144 TI - Neurotransmitter enzyme and receptor regulation: a look back. AB - 1. This report is a tribute to Dr. Julius Axelrod, in whose laboratory the author worked as a research associate from 1971-1974. 2. Work on the regulation of dopamine beta-hydroxylase and phenylethanolamine N-methyl transferase is reviewed, particularly studies showing how transynaptic and hormonal factors regulate enzyme synthesis and degradation respectively. 3. Work taking place in the author's laboratory is reviewed with respect to the influence Dr. Axelrod had on the author and his subsequent work. PMID- 3042143 TI - Nerve growth factor: cellular localization and regulation of synthesis. AB - 1. The role of nerve growth factor (NGF) as a retrograde messenger between peripheral target tissues and innervating sympathetic and neural crest-derived sensory neurons is supported by the observations that (a) the interruption of retrograde axonal transport has the same effects as the neutralization of endogenous NGF by anti-NGF antibodies and (b) the close correlation between the density of innervation by fibers of NGF-responsive neurons and the levels of NGF and mRNANGF in their target organs. 2. In situ hybridization experiments have demonstrated that a great variety of cells in the projection field or NGF responsive neurons is synthesizing NGF, among them epithelial cells, smooth muscle cells, fibroblasts, and Schwann cells. 3. The temporal correlation between the growth of trigeminal sensory fibers into the whisker pad of the mouse and the commencement of NGF synthesis initially suggested a causal relationship between these two events. However, in chick embryos rendered aneural by prior removal of the neural tube or the neural crest, it was shown that the onset of NGF synthesis in the periphery is independent of neurons, and is controlled by an endogenous "clock" whose regulatory mechanism remains to be established. 4. A comparison between NGF synthesis in the nonneuronal cells of the newborn rat sciatic nerve and that in the adult sciatic nerve after lesion provided evidence for the important regulatory role played by a secretory product of activated macrophages. The identity of this product is currently under investigation. PMID- 3042145 TI - Protein-carboxyl methylation in adrenal medullary cells. AB - 1. The protein-carboxyl methylating system has been studied in adrenal medullary cells either using disrupted cell components or with intact cells. Whereas the enzyme protein-carboxyl methylase (PCM) is cytosolic, the majority of its substrates is on or within chromaffin granules. With intact granules, methylation of surface proteins results in solubilization of membrane proteins. 2. Membrane PCM substrates have been identified as two proteins with apparent molecular weights of 55,000 and 32,000. Among the substrates located inside the granules, the chromogranins are excellent substrates, while dopamine beta-hydroxylase is poorly methylated. 3. Under physiological conditions, stimulation of the splanchnic nerve results in an increase in adrenal medullary protein-methyl ester formation as well as in an augmented methanol production. With adrenal medullary cells in culture, carboxyl-methylated chromogranin A is detected in mature chromaffin granules between 3 and 6 hr after labeling. Methylated chromogranins are secreted concomitantly with catecholamines following cholinergic stimulation. 4. These data coupled with those of Chelsky et al. (J. Biol. Chem. 262:4303-4309, 1987) on lamin B suggest that PCM methylates residues other than D-aspartyl and L isoaspartyl in proteins. They further suggest that methylation may occur on nascent peptide chains before they are injected into the rough endoplasmic reticulum. PMID- 3042146 TI - [Thomayer's medical cultural history]. PMID- 3042147 TI - [The last Czech ennoblement]. PMID- 3042148 TI - [The importance of determining immunoglobulins, C3 and C4 complement fractions, carcinoembryonic antigen and beta 2 microglobulin in patients with pleural effusion]. PMID- 3042149 TI - [25 years of the Second Surgical Clinic of the Medical School Hospital in Olomouc]. PMID- 3042150 TI - Post-lumbar-puncture headache: the significance of body posture. A controlled study of 300 patients. AB - In this single-blind, randomized study of post-lumbar-puncture headache (PPH) in 300 neurologic inpatients the significance of body posture after lumbar puncture (LP) was evaluated. Immediate mobilization was compared with bed rest for 6 h (3 h prone followed by 3 h supine posture). Contrary to the widely held belief, this investigation did not show significant differences between recumbent and ambulant patients as to frequency of PPH in the total material (39% versus 35%) or when men (31% versus 29%) and women (48% versus 41%) were evaluated separately. Headache associated with nausea was significantly more frequent in the recumbent than in the ambulant patients both in the total material (23% versus 13%) and in women (35% versus 16%). Thus, immediate mobilization seems to be preferable after LP. PMID- 3042151 TI - Duchenne muscular dystrophy: deficiency of dystrophin at the muscle cell surface. AB - Dystrophin is the altered gene product in Duchenne muscular dystrophy (DMD). We used polyclonal antibodies against dystrophin to immunohistochemically localize the protein in human muscle. In normal individuals and in patients with myopathies other than DMD, dystrophin was localized to the sarcolemma of the fibers. The protein was absent or markedly deficient in DMD. The sarcolemmal localization of dystrophin is consistent with other evidence that there are structural and functional abnormalities of muscle surface membranes in DMD. PMID- 3042152 TI - Time of replication of yeast centromeres and telomeres. AB - The time of replication of centromeres and telomeres of the yeast S. cerevisiae was determined by performing Meselson-Stahl experiments with synchronized cells. The nine centromeres examined become hybrid in density early in S phase, eliminating the possibility that a delay in the replication of centromeres until mitosis is responsible for sister chromatid adherence and proper chromosome segregation at anaphase. The conserved sequence element Y', present at most telomeres, replicates late in S phase, as do the unique sequences adjacent to five specific telomeres. The early and late replication times of these structural elements may be either essential for their proper function or a consequence of some architectural feature of the chromosome. PMID- 3042153 TI - Core sequence of two separable terminus sites of the R6K plasmid that exhibit polar inhibition of replication is a 20 bp inverted repeat. AB - The DNA replication terminus (terR) of the R6K plasmid located on a 216 bp Alul fragment (Alu216) can block progress of the DNA replication fork. We previously developed an electrophoresis assay that allows detection of terminus activity on any DNA fragment cloned in the pUC vector. For precise identification of terR, we tested Alu216, its subfragments, and synthetic oligonucleotides by this assay. We found terR to be composed of a pair of separable sites, terR1 and terR2, each of which can block the DNA replication fork traveling in a specific but not the opposite direction. Both terR sites were composed of 22 nucleotides containing the repeated 20 bp sequence 5'-TAGTTACAACAC(A or T) CAA(G or T) AGA-3', located 73 bp apart in the inverted position of Alu216. A DNA homology search suggested that the R6K plasmid and the E. coli chromosome share a common termination system. PMID- 3042154 TI - The mechanics of winding and unwinding helices in recombination: torsional stress associated with strand transfer promoted by RecA protein. AB - Homologous recombination usually involves the production of heteroduplex DNA, DNA containing strands contributed from two different duplexes. RecA protein of E. coli can promote the formation of heteroduplex DNA in vitro by the exchange of DNA strands between two helical structures, duplex DNA and a helical recA nucleoprotein filament containing a single strand of DNA. Complete unwinding of the parental duplex and the rewinding of one strand with a new complement requires rotation of the helical structures about one another, or about their respective longitudinal axes. The observations described here demonstrate an association of torsional stress with strand exchange, and suggest that exchange is accomplished principally by concomitant rotation of duplex DNA and the recA nucleoprotein filament, each about its longitudinal axis. PMID- 3042155 TI - Photosensitization of leukemic cells and normal bone marrow cells by 514 nm laser light and effects of laser light on migration inhibition and lymphokine response. AB - In a model for ex-vivo purging of bone marrow grafts, leukemic cells and normal bone marrow cells were treated with merocyanine 540 and exposed to 514 nm laser light. With this treatment, 99.9999% of leukemic cells were killed while 55% of the normal bone marrow cells survived. The deleterious effects of laser light alone in the absence of photosensitizer were not observed as determined by cell viability, cell migration, and response of target cells to human migration inhibition factor. These results indicate that laser light induced photodynamic therapy can be useful for ex-vivo autologous bone marrow purging without regard to the deleterious effects of laser light alone. PMID- 3042156 TI - Head and spine injuries in the young athlete. AB - Recent studies have shown a decrease in the mortality rates from head and neck injuries, especially in American football. This has resulted because of rule changes and their enforcement, equipment modifications, improved coaching and training techniques, and educational programs for coaches, trainers, and team physicians on the early recognition of head and neck injuries. However, morbidity data is not as complete, particularly as it applies to concussion, the most frequent type of head injury in contact sports. Questions on this condition that still need to be answered before a sound medical disposition can be made are the possible cumulative damage from repeated concussions, and whether one concussion renders a player more susceptible to a second. Presently, decisions on when to allow a football player to return to a game or participate in future contests are arbitrary and based primarily on the experience of the team physician. Data on the incidence, mechanisms, and prognosis of transient spinal cord signs and symptoms, such as spinal cord concussion and the central core symptoms, is also incomplete. What is the long-term prognosis for players who suffer frequent "burners"? Certainly, further studies are essential before these questions can be answered. Thus, the pioneer work of Richard Schneider needs to be continued. PMID- 3042157 TI - Shoulder and elbow injuries in the young athlete. AB - The skeletally immature athlete sustains upper extremity injuries unique to the epiphyseal plate, articular cartilage, musculotendinous units, and specific to the sport itself. Specific shoulder and elbow conditions can be predicted based on the biomechanics of the sport and age of the patient. In the young athlete, recognition of these unique injury patterns with early activity modification and treatment can prevent permanent deformity and functional disability. PMID- 3042158 TI - Hand and wrist injuries in children. AB - Hand and wrist injuries in children have steadily increased over the past decade as athletic participation by both boys and girls at an earlier age has been encouraged by parents and coaches. This article will cover the spectrum of hand and wrist injuries in children including ligamentous, tendinous, and bony injuries, as well as the increasing incidence of stress-related injuries in the upper extremities. PMID- 3042159 TI - Hip and pelvic injuries in the young athlete. AB - Hip and pelvic injuries are relatively rare in the young athlete. Contusions and musculotendinous sprains are the most common injuries about the hip and pelvis. Apophyseal avulsion fractures and stress fractures are the most frequently encountered skeletal injuries. Each of these entities can be successfully treated with guided physical therapy following conservative management with rest, anti inflammatory medications, and ice massage until the patient is pain free. Epiphyseal, diaphyseal, or pathologic fractures are rare entities that are secondary to violent trauma. These injuries are severe and often require operative intervention. Femoral neck fractures have a high rate of complications from avascular necrosis, nonunion, or malunion. Pelvic fractures have frequent associated genitourinary, abdominal, neurologic, and musculoskeletal injuries. Pathologic fractures are most commonly secondary to benign lesions, such as unicameral bone cysts, and less likely owing to malignancy. Finally, in children with hip pain during athletic activities, even with antecedent trauma, the sports clinician must screen for slipped capital femoral epiphysis, Perthes' disease, congenital subluxation of the hip, toxic synovitis, systemic neoplasia, or infectious process. PMID- 3042160 TI - The young athlete's knee: recent advances. AB - The understanding and treatment of the young athlete's knee has improved and continues to do so as arthroscopy and good long-term follow-up studies guide the process. In the management of epiphyseal fractures about the knee the presence of associated ligament injury and the high likelihood of a growth plate injury is now well documented. In the management of ligament injuries and internal derangements, arthroscopy has profoundly changed diagnosis and treatment. Pathology can be precisely identified and the appropriate treatment initiated to preserve an athletic career and possibly prevent degenerative arthritis in adulthood. Lastly, patellofemoral pain continues to be a difficult problem to treat, but a more scientific approach is evolving. This has been facilitated by a better understanding of the varied etiologies for anterior knee pain, and a trend away from classifying all such conditions as chondromalacia. PMID- 3042161 TI - Overuse injuries in the young athlete. AB - Overuse injuries are now well known to sports enthusiasts at any age or level of competition. The seeming explosion of overuse stress fractures of lower extremity bones in high-profile professional basketball players has brought about widespread media attention and a better understanding of the phenomenon of "overuse syndrome" by the public. However, the spectrum of overuse injuries in the child or adolescent athlete has only recently been recognized. These injuries can range from the permanent disability of osteochondritis dissecans of the elbow to the completely nonspecific "growing pains" of active youngsters. PMID- 3042162 TI - Acute soft-tissue injuries in the young athlete. AB - Although the incidence of soft-tissue injury is high in the young athlete, one must be constantly aware of the tendency toward epiphyseal and apophyseal injuries in individuals with open growth plates. After the diagnosis of a soft tissue injury (sprain, strain, or contusion) has been made, treatment must include an initial 24- to 48-hour period of RICE. Appropriate splinting may be required. Rehabilitation then proceeds aggressively, with early restoration of strength, flexibility, and joint range of motion. Prior to return to full athletic activity, the athlete should meet the criteria outlined in this article. Protective taping or bracing may be necessary upon return to full activity. The treatment of soft-tissue injuries should start with prophylaxis. All predisposing factors to the development of injury should be sought on preparticipation physical examinations and corrected prior to allowing the young athlete to compete. Using the program described as a guide to treating the injured athlete should result in early return to function with low recurrence rates of injury. PMID- 3042163 TI - Emergencies in sports: the young athlete. AB - Accidents are the leading cause of death in children over the age of 1 year. Owing to the prevalence of injury, especially sports related, the attending physician should always be alert for the plausibility of serious medical emergencies in the young athlete. PMID- 3042164 TI - Musculoskeletal assessment and training: the young athlete. AB - The primary purpose of a preparticipation physical examination is to identify risk factors that may predispose an athlete to physical and/or psychological injury. Inclusion of a physiological assessment complements the more traditional preparticipation health examination by contributing valuable information toward the specific physical strengths and weaknesses of the young athlete. This information when combined with the orthopedic and medical components of a preparticipation examination can dramatically enhance the safety of sports participation. PMID- 3042165 TI - Isolation and characterization of Saccharomyces cerevisiae mutants resistant to T 2 toxin. AB - T-2 toxin, a trichothecene mycotoxin, inhibits the growth of Saccharomyces cerevisiae. We have isolated nine spontaneous S. cerevisiae mutants resistant to this toxin. The mutants were distinguished from the wild type according to their degree of resistance to T-2 toxin on media with dextrose or glycerol as the carbon source. Generation time, mutation stability and level of cross-resistance to roridin A, another trichothecene, were determined for each mutant. The T-2 toxin resistant mutants were further characterized by subsequent tests involving cross-resistance and collateral sensitivity to chlorampenicol, neomycin, paromomycin, ethidium bromide and thiolutin. Mutants have been placed into three subgroups and the mechanism of T-2 toxin resistance in each group has been postulated. Mutant HK1 is the first S. cerevisiae isolate resistant to roridin A. One particular isolate, mutant HK11, carries a single recessive nuclear mutation. This mutation was termed ttt (for T-2 toxin resistant). PMID- 3042166 TI - MAL64c is a global regulator of alpha-glucoside fermentation: identification of a new gene involved in melezitose fermentation. AB - Maltase constitutive mutants at the MAL6 locus have been mapped to the newly identified regulatory gene MAL64c. We show here that MAL64c has in addition pleiotropic effects on sugar fermentation: MAL64c strains constitutively synthesize an alpha-methylglucosidase and can complement a new gene, MTP1, for the fermentation of melezitose and alpha-methylglucoside. MTP1, maps near MAL1, and either encodes a permease which transports melezitose, alpha-methylglucoside, and maltose or regulates the activity of such a permease. This work shows that MAL64c, a trans-acting regulatory gene, is a global regulatory gene affecting several different pathways of alpha-glucoside metabolism. PMID- 3042167 TI - Induced recombination and reversion of the CDC8 gene in relation to the cell cycle of yeast. AB - The induction of mitotic recombination in the CDC8 locus was studied in a diploid strain heteroallelic for cdc8 mutations (cdc8-1/cdc8-3); mitotic reversion was studied in strains cdc8-1/cdc8-1 and cdc8-3/cdc8-3. Conversion and reversion did not occur in those cells blocked at the S stage of the cell cycle by exposure to a nonpermissive temperature. In stationary phase cells irradiated just prior to exposure to temperature stress, the induction of recombinants was rather low and the induction of revertants was minimal. Conversely, a significant induction of cdc+ occurred in logarithmic phase cells subjected to the same treatment. Irradiation of synchronously dividing cultures revealed that intragenic recombination occurs at all three stages of the cell cycle-G1, S and G2. It was also found that UV-induced gene reversion can occur during the S and G2 stages, but not during the G1 stage of the cell cycle. PMID- 3042168 TI - Yeast cell viability under conditions of high temperature and ethanol concentrations depends on the mitochondrial genome. AB - Wine yeasts manifest simultaneously a high tolerance to ethanol, thermotolerance, and a high resistance to the mutagenic effects of ethanol on the mitochondrial genome. The transfer of mitochondria from these strains to laboratory yeasts demonstrate that this genome influences the above parameters, since thermotolerance, ethanol-growth tolerance, and the frequency of rho- mutants were either totally or partially modified in the laboratory recipient strain. When the death rate and the rate of formation of rho- mutants were measured under extreme conditions of inhibitory ethanol concentrations and high temperature, a perfect correlation was found between these parameters, and both of them were dependent on the strain of mitochondrial genome. Thus, the transfer of wine yeast mitochondria leads to a lower death rate, and a simultaneous increase in thermotolerance and ethanol tolerance in the recipient strain. These results demonstrate the role that viability plays under conditions of high temperatures and high ethanol concentrations. The greater stability of the rho+ phenotype shown by the wine yeast mitochondrial genome may be responsible for the increased viability conferred by these mitochondria. PMID- 3042171 TI - [How did children learn to walk in earlier times?]. PMID- 3042170 TI - [The importance of strict control of blood glucose at the onset and during the remission stage in newly diagnosed diabetic children]. PMID- 3042172 TI - [Prenatal diagnosis]. PMID- 3042169 TI - Behaviour of a replicating mitochondrial DNA sequence from Aspergillus amstelodami in Saccharomyces cerevisiae and Aspergillus nidulans. AB - An amplified sequence of mitochondrial DNA from a ragged (rgd) mutant of Aspergillus amstelodami has been shown to exist in multimeric circular form, suggesting that it is excised from the genome and can exist independently of it. This sequence has replicative (ARS) activity in Saccharomyces cerevisiae, and a subfragment responsible for this activity has been identified and sequenced. A homologous sequence from Aspergillus nidulans mtDNA also has ARS activity in S. cerevisiae. Both A. amstelodami and A. nidulans ARS elements have been incorporated into the integrative transformation vector pDJB1 and the derived vectors used to transform A. nidulans. Inclusion of the A. nidulans ARS element enhanced the transformation frequency 5-fold relative to pDJB1. No increase in transformation frequency was evident with the ARS element from A. amstelodami. The stability of transformants was variable but in comparison to pDJB1, ARS containing plasmids were mitotically unstable in A. nidulans. Although plasmid DNAs could be rescued in Escherichia coli from undigested transformant DNA, no freely replicating plasmids were detected by Southern hybridisation. PMID- 3042173 TI - [Pathomorphological and ultrastructural study on the rejection of human renal allografts]. PMID- 3042175 TI - [DNA hybridization in diagnostic pathology]. PMID- 3042174 TI - [Immunofluorescent distribution of basement membrane components and intermediate filament proteins in 10 cases of renal cell carcinoma]. PMID- 3042176 TI - [Aedes aegypti (L.): importance of its bioecology in the transmission of dengue and other arboviruses. II. Bibliography]. PMID- 3042177 TI - [Rift Valley Fever and phleboviroses in the Central African Republic]. AB - During 1984 and 1985, six Rift Valley Fever virus strains (RVF) were isolated in Central African Republic, among them five from human samples. Three strains were isolated in 1985 at the end of the rainy season, from sera of patients dead with severe jaundice with haemorrhagic syndrome, what could evoke a little outbreak. At this occasion, these RVF strains and the other strains of phleboviruses previously isolated in CAR, were antigenically compared by Immuno-fluorescent assay (IFA) and Complement Fixation Test (CF), using mice immuno ascitic fluids prepared against each strain. A lot of cross reactions were noted between the different strains, but all the RVF strains seem to have the same antigenic outline. PMID- 3042178 TI - [Chemoresistance of Plasmodium falciparum in Kinshasha. In vivo tests and chloroquine absorption]. AB - With the test in vivo and the bleeding dosage of chloroquine, authors report the rate of resistance of chloroquine to Plasmodium falciparum. 2.3% of Plasmodium falciparum in Kinshasa are resistant to chloroquine. PMID- 3042179 TI - [In vitro activity of various antimalarials (chloroquine, amodiaquine, quinine and mefloquine) against 32 isolates of Plasmodium falciparum in French Guiana]. AB - The in vitro susceptibility of various Plasmodium falciparum isolates obtained from patients infected in French Guiana was tested in a semi-microtest, using four antimalarial drugs. 90.6% of 32 isolates were resistant to chloroquine, 40% to amodiaquine and 17% to quinine. All the 17 isolates were susceptible to mefloquine. PMID- 3042180 TI - [Aedes aegypti (L.): importance of its bioecology in the transmission of dengue and other arboviruses. I]. AB - The bioecological parameters which are of special importance in the epidemiology of Dengue, Yellow Fever, and other arboviruses are discussed. Three levels are retained: the nature of Aedes aegypti-man contacts, the susceptibility of the mosquito to the pathogen and multiplication of the latter, and the transmission. The trophic preferences, the density variations, the daily survival rate, the egg diapause, and man influences are the main vector-dependent ecological factors. Temperature and genetical nature of viral and mosquito strains are particularly important in susceptibility and multiplication studies. Efficacy of the oral transmission is also temperature-dependent and mainly genetically determined. The true natural role of transovarial transmission is not yet well understood. Thus, the breaking up and/or prevention of epidemics would be possible only with a thorough knowledge of the relation between the above biological factors and the epidemiological situation. A list is provided of the naturally or experimentally Aedes aegypti transmitted arboviruses (103), protozoans (5) and filaria (20). PMID- 3042182 TI - Atypical malignant hyperthermia with persistent hyperkalaemia during renal transplantation. AB - A 35-year-old 110 kg male developed marked hyperkalaemia, hyponatraemia, hypercapnia and hyperthermia during living-related renal transplantation under anaesthesia with oxygen-nitrous oxide, isoflurane and muscle relaxation with atracurium. This is the first report of successfully treated malignant hyperthermia triggered by isoflurane during renal transplantation with early appearance and persistent (to 12 hours after surgery) electrolyte abnormalities. PMID- 3042184 TI - System for application of PEEP. PMID- 3042185 TI - Further characterization of a model of chronic endotoxemia in the rat: adenine nucleotide content in liver. AB - Male Sprague-Dawley rats (320-380 g) were treated with Escherichia coli endotoxin (ET) either acutely (3 and 4.5 mg/100 g b.w.,) or chronically (0.1 mg/100 g b.w./24 h, i.v. from subcutaneously implanted osmotic pumps). Control rats received only sterile saline. At 6 h after injection and after 30, 54, 126, and 198 h of continuous ET infusion, the animals were sacrificed, and adenine nucleotide content in liver was assayed. Acute ET treatment with either dose produced a slight decrease in ATP content (15% and 11% with 3 mg and 4.5 mg ET/100 g b.w., respectively) associated with an increase of ADP and AMP content and a fall of the energy charge (by 9.2% and 6.4%, with the two ET doses, respectively). Chronic ET treatment did not affect either of the measured parameters. However, in both saline- and ET-infused rats a decrease of ATP content paralleled by an increased level of ADP and AMP and by a fall of energy charge were seen when compared to acutely saline-injected rats. Such changes can be attributed to the surgical trauma associated with pump implantation. It is concluded that the energy status of the liver during endotoxemia cannot account for metabolic disturbances associated with this pathophysiological state. PMID- 3042183 TI - Acute epiglottitis in adults. PMID- 3042181 TI - Sickle cell states and the anaesthetist. PMID- 3042186 TI - Beneficial effects of the peptidoleukotriene receptor antagonist, SK&F 104353, on the responses to experimental endotoxemia in the conscious rat. AB - The purpose of this study was to examine the effects of the peptidoleukotriene receptor antagonist, SK&F 104353, on the responses to endotoxin in conscious male Sprague-Dawley rats. Administration of Salmonella enteritidis endotoxin (30 mg/kg i.v.; LD90) resulted in a decrease in the number of circulating platelets, leukopenia, an increase in hematocrit, and 0% survival at 24 hr. Pretreatment with SK&F 104353 (1 mg/kg, i.v. bolus followed by 3 mg/kg/hr, i.v. infusion for 6 hr) 5 min before injection of endotoxin produced steady state plasma drug levels of 1.6 micrograms/ml in naive animals and levels of approximately 3.4 micrograms/ml in endotoxemic animals (P less than 0.05). SK&F 104353 significantly attenuated the endotoxin-induced thrombocytopenia (P less than 0.05) but had no effect on either the endotoxin-induced early leukopenia or late leukocytosis. Additionally, SK&F 104353 significantly reduced the endotoxin induced hemoconcentration (P less than 0.05) and improved survival to 30% at 48 hr (P less than 0.05). A higher dose of SK&F 104353 (2 mg/kg, i.v. bolus followed by 10 mg/kg/hr, i.v. infusion for 6 hr) did not produce any further benefit. These data indicate clearly the pathophysiologic role of peptidoleukotrienes in endotoxemia and suggest SK&F 104353 could be useful for ameliorating some of the deleterious sequelae associated with this condition. PMID- 3042187 TI - The role of complement in glucan-induced protection against septic shock. AB - Previous studies from our laboratory have shown that glucan will significantly enhance survival, decrease bacteremia, maintain reticuloendothelial function, and reduce histopathology in a murine model of gram-negative septic shock [1]. The present study was undertaken to evaluate the role of complement in glucan enhanced protection against septic shock. AKR/J mice, which are congenitally C5 deficient, and ICR/HSD mice that were complement-depleted by treatment with purified cobra venom factor (CVF), were injected IP with glucan (50 mg/kg) on days 5 and 3 prior to IP challenge with 1 X 10(8) E. coli. Survival data indicated that glucan (p less than 0.05) increased survival in both C5-deficient and complement-depleted mice. Glucan prophylaxis resulted in a neutrophilic leukocytosis 8 h following E. coli challenge. However, glucan did not alter bone marrow proliferation. We conclude that, 1) glucan's protective effect on survival is not dependent on complement, 2) complement is not required for glucan-induced neutrophilic leukocytosis in this model, and 3) glucan does not enhance bone marrow proliferation in complement-deficient mice. PMID- 3042188 TI - Energy metabolism during chronic endotoxin infusion in the rat, with special reference to free fatty acid turnover. AB - We have studied various aspects of whole-body energy metabolism during chronic endotoxin infusion in the rat. In particular, we studied free fatty acid kinetics using bolus injections of 14C-palmitate. The general response to 7 days of chronic endotoxin infusion could be divided into an initial 3- to 4-day period of illness with loss of appetite, followed by rapid recovery. Using saline-infused, pair-fed controls, the following observations were made throughout the 7-day period of endotoxin infusion: 1) Plasma free fatty acids, glycerol and 3 hydroxybutyrate concentrations were reduced; 2) plasma glucose concentrations were elevated; 3) free fatty acid production rates were unaffected; 4) metabolic clearance of free fatty acids was unaffected on day 1 but was significantly greater on days 3 and 7; 5) haematocrits and plasma volumes were elevated. The results suggest that the observed changes in circulating free fatty acid concentrations are more likely to be a consequence of haemodynamic alterations than of metabolic alterations. PMID- 3042189 TI - Confirmation and certainty in toxicology screening. AB - Confirmation of presumptive positive urine drug screens, necessary to minimize the reporting of false-positive results, can be costly and time-consuming. The predictive value model can be used to select the confirming tests and to calculate the confidence of the result. The predictive value of a test result is the probability, based on the sensitivity and specificity of the test, that the result is a true positive or a true negative. The predictive value model applied to toxicology screening tests for drugs of abuse showed that prevalence, in addition to sensitivity and specificity, was the factor controlling the confidence level of a result. For example, the predictive value of a positive result for a screening test that has a sensitivity of 99% and a specificity of 99%, applied to screening in a population with a prevalence of 1% is 0.50; for a prevalence of 10%, it is 0.92. Confirmation with a second, chemically independent, test of equal sensitivity and specificity increases the predictive value to 0.99. PMID- 3042190 TI - A computerized classification technique for screening for the presence of breath biomarkers in lung cancer. AB - A simple computer-based screening technique has been developed for classifying human expired air components into 16 chemical classes, based on empirical formulas. The sort procedure was developed to simplify the screening of the composition of expired air samples by sorting all components into chemical classes and classifying components at the greater than 75% and greater than 90% occurrence levels. Both occurrence-rate components are then evaluated as diagnostic markers in a discriminant function model for their ability to detect lung cancer. Of the 386 components detected in the gas chromatography/mass spectrometry (GC/MS) data files, 45 components were present at the greater than 75% occurrence level and 28 components at the greater than 90% occurrence level. Thus, this preliminary sort routine, performed by using a simple macro program installed into a standard personal-computer spread-sheet, greatly reduces the amount of data required for statistical treatment. Such a sort routine can also be applied as easily to other complex GC/MS data files for the purpose of data reduction. PMID- 3042192 TI - Effect of heat inactivation on results of HIV antibody detection by Western blot assay. PMID- 3042191 TI - Sensitive, rapid procedure for time-resolved immunofluorometry of lutropin. AB - In this new immunofluorometric method for quantification of lutropin in serum, the "sandwich" principle is combined with time-resolved fluorescence measurements, with the europium chelate 4,7-bis(chlorosulfophenyl)-1,10 phenanthroline-2,9-dicarboxylic acid (BCPDA) used as label. A monoclonal antibody to the alpha-subunit of lutropin is adsorbed onto the walls of white-opaque microtiter wells to form the solid-phase capture antibody, and a biotin-labeled soluble monoclonal antibody is used for antigen quantification. The detection system is completed with streptavidin, which has been linked to a protein bulking agent labeled with multiple BCPDA residues. In the presence of excess europium, the fluorescence of the final complex attached to captured lutropin molecules is measured on the dried solid phasse with an automated time-resolved fluorometer. The assay can be performed as a rapid (less than 60 min incubation) or regular (150 min incubation) procedure. The rapid assay is well-suited for routine daily monitoring of increasing or ovulatory lutropin concentrations; the regular assay, with its greater sensitivity (0.5 int. unit/L), is a practical procedure for lutropin measurements in hyposecretory states. The assay measures up to 240 int. units/L, and results compare well with those by a commercially available radioimmunoassay, an immunoradiometric assay, and another time-resolved immunofluorometric procedure. PMID- 3042193 TI - Dietary treatment of hyperlipidemia. PMID- 3042194 TI - Cardiovascular risk factors in children and early prevention of heart disease. AB - Clinical and anatomical observations from the Bogalusa Heart Study over a 15-year span provide data on cardiovascular risk factors and the early natural history of arteriosclerosis. These studies established that: cardiovascular risk can be predicted in early life; interrelationships of risk factors in children are similar to those observed in adults; and concentrations of serum lipoproteins change during sexual maturation. Strategies involving the general population and high-risk groups should be considered to help reduce atherosclerosis and coronary artery disease. Active modification of risk factors in the general populace would include using such methods as screening, education, and mass-media campaigns. Educational promotion ("Heart Smart") has already been implemented in the New Orleans area, focusing on using the total school environment to teach cardiovascular health and to encourage children to adopt desirable lifestyles. Inclusion of cardiovascular health education into general educational studies of children should be a major objective of the future. PMID- 3042195 TI - Drug therapy of hypercholesterolemia. AB - The rationale for drug treatment of patients with hypercholesterolemia is that sustained reductions in the plasma concentrations of atherogenic lipoproteins will retard the development of atherosclerosis. Drug therapy should be initiated only after the exclusion of secondary factors and after an adequate trial of dietary therapy has failed to lower plasma cholesterol to a satisfactory level. The bile-acid sequestrants (cholestyramine and colestipol), nicotinic acid, and lovastatin are the most effective drugs for use in patients with primary hypercholesterolemia; these agents reduce total and LDL cholesterol concentrations by 15-35%. For patients with high concentrations of LDL cholesterol who concurrently have hypertriglyceridemia, nicotinic acid is the drug of choice, and bile-acid sequestrants are contraindicated. Combined therapy with drugs that have different mechanisms of action can be effectively utilized in the treatment of patients with severe hypercholesterolemia; combinations involving lovastatin, nicotinic acid, and bile-acid sequestrants are the most effective. PMID- 3042196 TI - Profiles of apolipoproteins and apolipoprotein B-containing lipoprotein particles in dyslipoproteinemias. PMID- 3042197 TI - The pathogenesis of atherosclerosis. PMID- 3042198 TI - Apolipoproteins and lipoproteins in human plasma: an overview. PMID- 3042199 TI - The epidemiology of atherosclerosis in 1987: unraveling a common-source epidemic. AB - Epidemiologists studying the risk factors of coronary heart disease, as indicated by a heart attack, often presume that these risk factors combine in a susceptible host to increase the risk of the heart attack. The "biological" basis for the interaction of the key risk factors is often not considered in the analysis. A more rational model for the study of clinical heart disease, stroke, and peripheral vascular disease would be to separate the epidemiology of atherosclerosis from that of the clinical event. In the past, this has been extremely difficult because of the absence of techniques for the measurement of atherosclerosis in vivo, especially in well-defined populations. However, in recent years, greater emphasis on the quantification of atherosclerosis and its relationship to specific risk factors has improved our ability to study the underlying pathology that is atherosclerosis. Atherosclerosis is an example of a common-source epidemic. The environmental agent is the intake of cholesterol and saturated fat. The interaction of specific dietary factors and genetic determinants is the key to the evolution of atherosclerosis. There are marked variations in the evolution of the disease in different vascular beds as well as among individuals exposed to similar environmental factors. In the future, we will probably be able to study atherosclerosis as a continuous variable in populations and to relate the rate of progression, the extent of disease at any point in time, and topical distribution of atherosclerosis within individual vascular beds to specific genetic and environmental determinants. PMID- 3042200 TI - Cholesterol and risk of coronary heart disease and mortality in men. AB - Extensive data implicate cholesterol in the atherosclerotic process responsible for coronary disease. Of the atherosclerotic disease outcomes, serum cholesterol is most strongly related to coronary disease. A significant relationship of serum cholesterol to all clinical manifestations of coronary heart disease has been demonstrated in the Framingham Study, after adjusting for coexistent risk factors. Cholesterol and blood pressure exert similar influences on the occurrence of coronary heart disease. Risk of coronary heart disease associated with serum cholesterol is continuous, graded, and strong, with ideal values for cholesterol probably in the 130-190 mg/dL range. The impact of serum cholesterol diminishes with advancing age, but the predictive value of cholesterol is restored when fractionated into its atherogenic LDL and protective HDL components. The predictive value of total cholesterol in serum at all concentrations, including values less than 200 mg/dL, can be enhanced by taking HDL cholesterol into account. The total/HDL cholesterol ratio is a practical, efficient means for evaluating the joint effect of the two-way cholesterol traffic. Other cardiovascular risk factors such as blood pressure, glucose, cigarette smoking, fibrinogen, and left ventricular hypertrophy markedly influence the risk associated with measured concentrations of serum cholesterol. In correcting hypertension or diabetes, lipid values are an important consideration in determining the urgency, type, and efficacy of treatment used. In contrast to coronary mortality, rates of overall mortality show a quadratic relationship to total cholesterol in serum, with excessive mortality at concentrations greater than 160 mg/dL. PMID- 3042201 TI - Cholesterol, lipoproteins, and coronary heart disease in women. AB - In the United States, coronary heart disease is the major cause of death and disability in women and in men. Despite this, little is known about the risk factors, including cholesterol and lipoprotein concentrations, for coronary disease in women. In this paper we review the determinants of cholesterol and lipoprotein concentrations in women, assess whether values for total cholesterol and lipoproteins (HDL and LDL) are associated with the occurrence of coronary heart disease in women, and evaluate the evidence that suggests that modifying the concentrations of lipids in women is associated with changing the risk of coronary disease. Besides genetic determinants, dietary cholesterol, dietary fat, total caloric intake, alcohol consumption, cigarette smoking, and physical activity are known to influence concentrations of lipids in women. Some of the strongest determinants of cholesterol and lipoprotein concentrations in women are sex hormones, including estrogen and progestin. Exogenous use of both of these hormones markedly influences HDL and LDL cholesterol; additional evidence suggests that endogenous sex hormones also influence lipid and lipoprotein concentrations. The few studies that have examined the association of total cholesterol with coronary heart disease occurrence and mortality in women have consistently shown that (a) women have much lower rates of coronary heart disease than men at the same values for cholesterol, and (b) clearly elevated risk for coronary heart disease in women is evident only at relatively high values of total cholesterol (i.e., greater than 260 mg/dL). There also appears to be an age effect, with total cholesterol concentrations being more predictive in older than in younger women. PMID- 3042202 TI - HyperapoB: a pleiotropic phenotype characterized by dense low-density lipoproteins and associated with coronary artery disease. AB - HyperapoB, a lipoprotein phenotype characterized by increased numbers of small, dense, low-density lipoproteins (LDL), is strongly associated with coronary artery disease (CAD). Patients with hyperapoB may be normolipidemic, hypertriglyceridemic, or, when the number of LDL particles increases sufficiently, hypercholesterolemic. Concentrations of high-density lipoprotein (HDL) and its major apolipoprotein, apo A-1, are often low in plasma of patients with hyperapoB. The increased number of dense LDL in hyperapoB is due to increased LDL synthesis, secondary to increased synthesis of very-low-density lipoproteins (VLDL) and apo B. HyperapoB may be a dominant trait, although the existence of a common recessive allele at a very high frequency has not been excluded. The expression of hyperapoB appears delayed, but the phenotype is commonly found in children referred to specialty lipid clinics because of a family history of premature CAD. Published data suggest a biochemical, genetic, and metabolic relationship between hyperapoB, familial combined hyperlipidemia, and the dense LDL subclass patterns described in Mormon families. The biochemical and genetic basis for the overproduction of VLDL apo B is under further study, both molecular investigations of the apo B gene and studies of free fatty acid, triglyceride, and HDL metabolism. PMID- 3042203 TI - Lipoprotein subspecies and risk of coronary disease. AB - This review summarizes physical and chemical properties of major subspecies of very-low-, low-, intermediate, and high-density lipoproteins. Hypotheses regarding the metabolic origins of these subspecies and evidence for their associations with risk of coronary artery disease are presented. PMID- 3042204 TI - Reliability of lipid, lipoprotein, and apolipoprotein measurements. AB - The National Heart, Lung, and Blood Institute national awareness program on cholesterol and heart disease has placed new demands on laboratorians to utilize and perform more reliable measurements of lipids, lipoproteins, and apolipoproteins. The general public's awareness and the clinicians' concerns about the reliability of laboratory testing make it paramount that the analytical problems and issues are identified and solutions are provided to increase the current state of reliability of the measurement of these blood constituents. To accomplish this, the initial step is to assess the current state of reliability of lipid, lipoprotein, and apolipoprotein measurements in the clinical laboratories. Accuracy and precision of measurements of total cholesterol, triglycerides, high-density lipoprotein cholesterol, and apolipoproteins A-I and B are extensively discussed, and general as well as some specific recommendations are provided for some of the apparent problems. PMID- 3042205 TI - Genetics and abnormalities in metabolism of lipoproteins. PMID- 3042206 TI - Standardization of lipid, lipoprotein, and apolipoprotein measurements. AB - Accurate laboratory measurement of serum cholesterol has become a national public health priority. National proficiency testing surveys indicate that laboratory inaccuracy in cholesterol testing is more of a problem than precision. Like precision, accuracy is a function of multiple pre-analytical and analytical sources of variation. Controlling pre-analytical sources of variation helps minimize such sources of variation as intraperson biological, behavioral, and clinical differences, and variations caused by sample collection, handling, and shipping. Standardization of analytical sources of variation helps to achieve and maintain desirable analytical performance, accurate reporting, and correct interpretation of a reported cholesterol result. The intraperson total variation in lipoproteins and their constituents is of primary interest when one is interpreting a single result or a series of results from a single person. The mean of multiple specimens from the same person is required if one is to obtain an accurate value for intraperson total cholesterol and minimize pre-analytical sources of variation. Standardizing analytical sources of variation in some instrument systems requires standardizing results by using "fresh" patients' specimens. PMID- 3042207 TI - Thyroid cancer. PMID- 3042208 TI - Fat graft myringoplasty--a prospective trial. AB - A prospective trial to determine the success rate of the fat graft myringoplasty technique is reported. A success rate of 76% overall was attained at review 1 year postoperatively with increased success for smaller perforations of the tympanic membrane. PMID- 3042209 TI - Rheumatoid arthritis: otorhinolaryngological manifestations. PMID- 3042210 TI - Tolerance of engrafted donor T cells following bone marrow transplantation for severe combined immunodeficiency. AB - Patients transplanted for the treatment of severe combined immunodeficiency (SCID) frequently develop a unique state of split lymphoid chimerism. Such patients have T cells of donor origin, and non-T cells which are predominantly or exclusively of host origin. We have studied the reactivity of engrafted donor T cells to host and/or donor antigens in 12 patients transplanted for SCID, focusing on the characteristics of the tolerance to host and/or donor MHC antigens observed in nine of these patients who were recipients of T-cell depleted, haploidentical parental bone marrow. In both proliferative and cytolytic assays, engrafted, donor-derived T cells were shown to be selectively nonreactive to histoincompatible host cells. This tolerance could not be ascribed to cells with suppressive activity in the engrafted T-cell population. T cells from a subset of patients, however, exhibited proliferative but not cytolytic reactivity to donor peripheral blood mononuclear cells. The responding cells were shown to be donor-derived CD3+ cells and were predominantly reactive to B-cell fractions from the donor. Two patients who received transplants from each parent in sequence engrafted T cells from one parent and had non-T cells of host, paternal, and maternal origin. The engrafted T cells proliferated weakly to B cells from the other parent, but were tolerant in cytolytic assays. Donor anti donor reactivity was seen only in haploidentical split chimeras who had not been treated with cytotoxic drugs prior to T-cell engraftment. This proliferative reactivity toward donor may be due to an absence of donor derived Ia+ antigen presenting cells resident in the thymus of SCID patients at the time when the T cell repertoire is being shaped. PMID- 3042211 TI - The relationship between macrophages and C5b-9 complement complexes in human atherosclerosis. AB - The relationship between macrophages and the terminal C5b-9 complement complexes was investigated in human arteries affected with atherosclerosis by using monoclonal antibodies and indirect immunoperoxidase, immunogold silver staining, and double-labeling immunohistochemical techniques. Macrophages were found in all the atherosclerotic arteries as immunoreactive deposits with a nucleus, considered as intact cells, or without a nucleus, considered as cell remnants. The double-labeling technique shows C5b-9 deposits partially colocalized on the intact macrophages or on the cell debris of macrophage origin. These data suggest that C5b-9 complement complex may be formed on activated or dying macrophages with subsequent promotion of inflammatory events and progression of the atherosclerotic lesions. PMID- 3042212 TI - Western blot analysis of serum antibody reactivity with human melanoma cell antigens in alopecia areata and vitiligo. AB - Antibody reactivity to melanocyte-derived cells was investigated in patients with alopecia areata or totalis by use of Western blot analysis of detergent solubilized membrane antigens of a human melanoma cell line, M14. Reactivity was detected in the sera of 9 of 27 alopecia areata or totalis patients, 8 of 13 vitiligo patients, and 6 of 24 normal control subjects. Significant differences between patient and control sera were found in the number and distribution of antibody specificities detected. In vitiligo sera, there was an increased prevalence of reactivity to a melanoma antigen of 52,000 mol wt. In contrast, the predominant specificities in alopecia areata sera were for antigens of 74,500 and 70,800 mol wt, and the majority of positive sera were from patients with total hair loss. These findings suggest that autoreactivity to pigmented cells occurs in certain patients with alopecia areata or totalis. PMID- 3042213 TI - Characterization of the molecules involved in thymocyte thymic epithelial cell adhesion. AB - Cell to cell adhesion is important for mechanisms of cellular recognition, growth, and differentiation. The identification of molecules involved in these interactions is necessary in order to understand the molecular basis of these processes. We have previously described the development of two different thymic epithelial cell lines (TECS and TECL). Using an assay with radiolabeled thymocytes we found that thymocytes can adhere specifically to these thymic epithelial cells. This adhesion is trypsin sensitive, suggesting that involvement of specific cell surface proteins. In the present study we further characterize and begin identification of the molecules involved in this interaction. We found that thymocyte binding to thymic epithelial cells requires the presence of Ca2+ and Mg2+ and is mediated by a molecule greater than 10,000 MW. Also, we identified several antibodies which inhibit the adhesion of thymocytes to TECS. The membrane nature of the molecule mediating this interaction was confirmed by the ability of thymocyte membranes to block the inhibitory antibodies. PMID- 3042214 TI - Suppression of glomerular IgA deposition in ddY mice through periodic injections of lipopolysaccharides. AB - We investigated the role of lipopolysaccharides (LPS) in glomerular IgA deposition of ddY mice. The incidence of IgA deposition was lower in the LPS injected mice at 10 and 13 months of age as compared with that of their age matched controls (10 months, LPS 20% vs control 60%; 13 months, LPS 14% vs control 100%). Serum levels of IgA were lower in the LPS-injected mice than in the control mice. There were no appreciable differences in the percentage value of Thy-1+ cells, the ratio of Lyt-1+/Lyt-2+ cells, mitogenic responses, or IL-1 activities between the LPS-injected and the control mice. On the other hand, the IgA responses of spleen and Peyer's patch cells in the LPS-injected mice were lower than those observed in the control mice. These results suggest that the persistent use of LPS directly suppresses gut-associated lymphoreticular tissue (GALT), and that the lower IgA response in GALT induced by LPS causes suppression of glomerular IgA deposition in ddY mice. PMID- 3042215 TI - Metabolic effects of continuous subcutaneous insulin infusion: evidence that a rise and fall of portal vein insulin concentration with each major meal facilitates post-absorptive glycemic control. AB - Eighteen lean adult volunteers with insulin-requiring diabetes mellitus attempted to achieve normoglycemia using continuous subcutaneous insulin infusion (CSII) or conventional insulin therapy (CIT) in a randomized crossover trial of 68 +/- 2.5 weeks (mean +/- SEM) duration. As reported (Diabetes Care 8: 447-55, 1985) the group with absent to low beta-cell function (C-peptide negative, n = 11) attained mean post-absorptive normoglycemia only during CSII vs CIT (p less than 0.05). Only following CSII was this without change in post-absorptive serum triglyceride concentrations (-4 +/- 5.6 vs 12 +/- 4.7 mg/dl; -0.04 +/- 0.6 vs 0.14 +/- 0.05 mM, p less than 0.05) or body weight (0.01 +/- 0.02 vs 0.05 +/- 0.01 kg/week, p less than 0.05). In the group with glucagon stimulated serum C-peptide 100-400 pmol/L (C-peptide positive) responses to CSII or CIT were equal. As total daily insulin dosage (0.05 +/- 0.04 U/kg/day) was the same under all conditions, to explain the efficacy of CSII, glucoregulatory hormone responses were examined. Pre- and post-test breakfast serum free immunoreactive insulin and plasma glucagon concentrations were essentially unaffected by C-peptide or treatment status. Erythrocyte 125I-insulin binding was decreased in the C-peptide negative group only during CSII (8.6 +/- 0.5 vs 10.1 +/- 0.7%, p less than 0.005); C peptide positive group receptor binding was consistently low (8.2 +/- 0.8, 8.4 +/ 0.9%). During CIT using intermediate-acting insulin post-lunch peripheral venous insulin failed to rise (p less than 0.05), but in the C-peptide positive group, on the basis of C-peptide responses to breakfast an undetected rise and fall of portal venous insulin was assumed to coincide with each meal. Thus, only during CIT in the C-peptide negative group, which received on average 6.4/wk/subject fewer pre-meal regular insulin boluses (p less than 0.01), was the frequency of meal-related change in portal insulinemia decreased. Consistent meal-related fluctuations in portal insulinemia inherent in CSII hepatocytes sensitized by a post-receptor mechanism to the suppressive effects of insulin on glucose output and thus were indirectly responsible for the observed improvement in glycemic control and lipid metabolism in the C-peptide negative group. PMID- 3042216 TI - Fate of grainhandlers with bronchial hyperreactivity. AB - Bronchial reactivity to methacholine and to grain dust were determined in a group of grainhandlers whose spirometry was being studied prospectively. Six workers showed specific bronchial responses to grain dust (indicative of occupational asthma), 21 did not show reactivity to grain dust but had bronchial hyperreactivity to methacholine and 40 had neither. Those with hyperreactivity were older, had lower FEV1 at initial examination and were more likely to experience a significant decline in FEV1 over a single working shift. Over 6 years of follow-up, those with hyperreactivity were not more likely to leave work and although they showed a more rapid decline in lung function over the period, because of the small number of individuals studied the difference between those with and without bronchial hyperreactivity was not significant. Among those with hyperreactivity to methacholine, there was a significant positive association between change in FEV1 over a single workshift and change in FEV1 over six years of followup, not seen in those without hyperreactivity to methacholine. Moreover, reactivity to methacholine was a stable observation over 6 years in the majority of studied individuals. We conclude that in the presence of bronchial hyperreactivity to methacholine, change in FEV1 over a single workshift more precisely predicts the trend in FEV1 over time in grainhandlers. PMID- 3042217 TI - Molecular analysis of genetic disorders. PMID- 3042218 TI - The role of cordocentesis in fetal diagnosis. PMID- 3042219 TI - Chorionic villus sampling. PMID- 3042220 TI - Genetic amniocentesis at 14 weeks or less. AB - The recent clinical trials indicate that EA is feasible. The use of ancillary tools, such as ultrasound, requires only a slight modification of the previously established techniques for MA. The amniotic fluid can be obtained and cultured. Combining the reported populations sampled, the procedural and cytogenetic failure rate were 2.0% and 0.3% respectively. The necessary information for prenatal diagnosis in the situations listed previously can be obtained. The one exception to this would be the patient at increased risk for NTD. As noted above, diagnostic levels for AFAFP have been established beginning at 13 weeks gestation. Therefore, amniocentesis before 13 weeks gestation should not be considered an option for these patients. The majority of the findings in these "pilot" studies have been promising, but the issue regarding the safety of EA has not been answered. Harrison et al. theorized that the function of the physiology hydramnios was to "provide a distention growth stimulus to the uterus ... and conversely to share in the maintenance of uterine inhibition." The amniotic fluid is constantly being replaced, having a complete turnover approximately every 3 hours. Does the reduction in volume, for even a short period of time, change the "distensive forces" enough to increase the pregnancy loss rate? Later publications have associated MA with increased rates of congenital orthopedic anomalies and neonatal pulmonary compromise. The true existence of these complications continues to be debated in the literature. Two of the authors have indicated that none of the reported neonates exposed to EA have had such anomalies.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3042221 TI - Magnetic resonance imaging in prenatal diagnosis. PMID- 3042222 TI - The role of the newer antimicrobial agents in obstetrics and gynecology. AB - The new antibiotics include interesting compounds--extended-spectrum cephalosporins and penicillins, combinations of older antibiotics plus beta lactamase inhibitors, and new classes such as the monobactams and fluoroquinolones. In addition to extended spectrums, some of these compounds offer more favorable kinetics, less toxicity, or decreased cost. Several general conclusions might help place this array of new antibiotics in a useful clinical perspective. The newer antibiotics discussed here are no more effective than what is currently used. However, several have very good in-vitro activity against pelvic pathogens and are reasonable single-agent therapy in mild to moderate postpartum and postoperative infections. These antibiotics include cefoxitin, cefotetan, and piperacillin. Although moxalactam has good activity, its use is limited by concerns regarding bleeding disorders. Cefotaxime and cefoperazone have somewhat less favorable spectra, especially against anaerobes, yet in limited clinical trials are as effective as those cited above. Penicillin/beta lactamase inhibitor combinations are currently being evaluated and appear to be reasonable choices. Although imipenem has an excellent in-vitro spectrum, it should probably be reserved for resistant cases. Aztreonam offers an alternative to gentamicin. However, in view of its greater cost, its use should be limited to patients in whom renal toxicity is a concern. For serious infections, combination therapy with clindamycin and gentamicin is our preference. For pelvic inflammatory disease, none of the agents is recommended as sole therapy due to the lack of coverage for chlamydia and frequent suboptimal coverage for anaerobes. Many of these agents have been effective for prophylaxis, but none has been shown to be superior to the older, less expensive agents such as cefazolin. Although many are effective in single doses, it is also likely that cefazolin is equally effective as a single dose. In addition, while most of the newer antibiotics are resistant to beta-lactamases themselves, they may induce their formation. This may ultimately result in limitations to the use of the relatively inexpensive prophylactic antibiotics. PMID- 3042223 TI - Treatment of Chlamydia, Mycoplasma, and group B streptococcal infections. PMID- 3042224 TI - The treatment of gonorrhea, syphilis, chancroid, lymphogranuloma venereum, and granuloma inguinale. AB - The "classic" venereal diseases are a diverse group of infections caused by established pathogens. Their epidemiologies are unique and relatively well understood. Likewise, the clinical presentations, although diverse, are well defined, allowing them to be readily suspected or diagnosed. Once the clinical diagnosis is considered, laboratory support methods are available to confirm the impression. This results in the provision of specific directed therapy that is curative. The spread of disease, however, must be considered, and the sexual partners must be identified, evaluated, and, where applicable, treated to prevent spread. The reinforcement of preventative methods is also an important aspect of controlling these infections. PMID- 3042225 TI - Use of prophylactic antibiotics in obstetrics and gynecology. PMID- 3042226 TI - The treatment of vaginitis: Trichomonas, yeast, and bacterial vaginosis. PMID- 3042227 TI - Treatment of urinary tract infections during pregnancy. AB - Urinary tract infections complicate 7-10% of pregnancies. Early detection and treatment of bacteriuria prevents most cases of pyelonephritis and its associated maternal and neonatal morbidity. The high risk for recurrence of bacteriuria and pyelonephritis mandate close monitoring of previously infected women and may require chronic suppressive therapy through the remainder of pregnancy. Acute cystitis is a distinct syndrome with a low recurrence rate and rarely associated with pyelonephritis. PMID- 3042228 TI - Treatment of postcesarean endomyometritis. AB - Improved understanding of the microbiology of postcesarean endometritis has dramatically changed the approach to its antibiotic therapy. Initial therapy should include broad-spectrum anaerobic coverage, including against all Bacteroides species, as well as gram-positive and gram-negative aerobic coverage. Moreover, ideally initial therapy should also include coverage of Chlamydia trachomatis. Furthermore, although the use of antibiotic prophylaxis for high risk patients undergoing cesarean section has significantly decreased their incidence of febrile morbidity, one must remember that prophylactic antibiotics have important bacteriologic effects that may limit the efficacy of monotherapy for the treatment of endometritis in prophylaxis failures. PMID- 3042229 TI - Current status of functional lower extremity surgery in adult spastic patients. AB - Ten percent of upper motor neuron spastic patients are candidates for functional surgery. If the main cause of the disability is abnormal muscle tone and not sensation, balance, cognition, or perceptual problems, good results from tendon lengthening or transfers are to be expected. The preoperative work-up should consist of examination, trial orthoses, nerve blocks, and most importantly dynamic electromyography. Releases or neurectomies are generally done in nonfunctional patients. Phenol blocks are a temporizing procedure during the phase of neurologic recovery. The patients all tolerated the anesthesia well, and the surgical risks are the same as in other lower extremity procedures. PMID- 3042230 TI - Management of the spastic upper extremity in the neurologically impaired adult. AB - Spasticity that interferes with upper extremity function is common in adults following stroke, brain injury, or anoxia. During the period of neurologic recovery definitive surgical procedures are avoided. Techniques to temporarily reduce spasticity include the implantation of a MicroPort reservoir and catheter for repeated branchial plexus blocks and phenol nerve blocks, which provide longer lasting relief of noxious muscle tone. Percutaneous blocks of the musculocutaneous and recurrent median nerves and motor point blocks of the pectoralis major, the brachioradialis, and forearm flexor muscles are easily performed at bedside. The motor branch of the ulnar nerve can be injected surgically with phenol to diminish intrinsic spasticity. When neurologic recovery has plateaued, hand placement can be improved in many patients following proximal release of the brachioradialis muscle and lengthening of the biceps and branchialis tendons. Hand function is enhanced by fractional lengthening of spastic wrist and finger flexors. Intrinsic spasticity must be addressed at the same time by phenol block or intrinic release. When extensor function is lacking, a tenodesis of the wrist extensors is helpful. The thumb-in-palm deformity requires proximal release of the thenar muscles as well as lengthening of the flexor pollicis longus. Contracture releases in the nonfunctional arm improve hygiene and ease care. PMID- 3042231 TI - Comparison between animal and human studies of skeletal muscle adaptation to chronic stimulation. AB - Functional electrical stimulation (FES) has recently emerged as a clinical tool for treatment of neuromuscular disorders. Chronic muscle stimulation, however, has long been used by basic scientists studying the details of the muscular adaptation process. Biochemical, morphological, and functional changes occur in skeletal muscle secondary to chronic stimulation. Chronic stimulation (12-24 hours per day for six weeks) results in a well-defined progression of changes in which a "fast" muscle becomes a typical "slow" muscle with a large decrease in force-generating capacity. On the other hand, clinical studies of FES have demonstrated muscle strengthening following treatment. An attempt is made to reconcile the results obtained in the two fields. PMID- 3042232 TI - The ultrastructure of the organic phase associated with the inorganic substance in calcified tissues. AB - An organic phase is closely associated with the mineral substance is all calcified matrices, where it can be demonstrated as crystal-bound proteins by biochemical methods and as crystal ghosts by electron microscopy. Interest in crystal ghosts derives chiefly from the observation that they have the same shape, arrangement, and orientation as inorganic crystallites, which suggests they may have a role in their formation. Histochemically, crystal ghosts of epiphyseal cartilage react with colloidal iron (pH 2.0), acidic phosphotungstic acid, ruthenium red, and a number of cations including calcium, barium, magnesium, lanthanum, strontium, and terbium chloride. Their reactivity is removed by methylation and only incompletely restored by saponification. Moreover, the crystal ghosts located at the periphery of the calcified areas contain vic-glycol groups, as shown by their reactivity with periodic acid-silver nitrate and periodic acid-thiosemicarbazide-osmium. All these reactions show that the crystal ghosts of epiphyseal cartilage contain acidic, probably sulfate groups and, at least initially, vic-glycol groups. Their reactivity decreases as the calcification process is completed. Although the available data are not sufficient to allow a full understanding of the nature and function of these structures, they seem to play an important role in calcification. The hypothesis is presented that crystal ghosts are preformed in calcifying matrices and are activated by the unmasking of the reactive groups in their polymeric molecule; the unmasked groups then link up with inorganic ions in such a way to form organic-inorganic structures the inorganic ions of which are arranged in an apatitelike configuration and the filamentlike shape of which is the same as that of the polymeric molecule. PMID- 3042233 TI - Functional neuromuscular stimulation neuroprostheses for the tetraplegic hand. AB - Functional neuromuscular stimulation (FNS) of the C5 and C6 tetraplegic upper extremity has been shown to be a valid clinical tool for restoring controlled movement in the paralyzed hand. The current clinical system consists of a shoulder position transducer controlling an external microprocessor-based stimulator, which excites paralyzed muscle via the peripheral nerve using percutaneous leads or a multichannel, implantable stimulator. Tendon transfer surgery of paralyzed but innervated muscle may augment the neurologically deficient upper extremity by allowing the substitution of stronger motors or the addition of new motors where flaccid paralysis (dennervation) eliminates the usual muscle from a grasp pattern. Sensory feedback in the form of machine state and cognitive information can be provided to the normally innervated C5 dermatome skin by subcutaneous electrodes. C5- and C6-level tetraplegics using FNS can independently perform single-hand manipulative tasks at a level similar to that of subjects with intact C7 roots, although they lack the elbow control. PMID- 3042234 TI - A clinical and histologic analysis of failed fresh osteochondral allografts. AB - Fresh small osteochondral allografts were implanted in 108 patients. A clinical and histologic analysis was undertaken in 18 patients whose grafts failed and were excised. Twelve of these patients had evidence of viable hyaline cartilage despite adverse conditions. If careful attention is given to patient selection, age, appropriate timing for correction of alignment, and meticulous technique associated with graft positioning, size, and thickness, then the incidence of clinical failure should be minimized. PMID- 3042235 TI - Preface to "A treatise on the nature of club-foot and analogous distortions; including their treatment both with and without surgical operation. Illustrated by a series of cases and numerous practical instructions". By W. J. Little, 1839. PMID- 3042236 TI - The effects of dimethyl sulfoxide on posttraumatic limb swelling and joint stiffness. A review and an experimental study in rabbits. AB - Dimethyl sulfoxide (DMSO) is an inorganic compound with many interesting in vitro properties, including the ability to scavenge oxygen-free radicals. DMSO has been used to treat a variety of clinical conditions, especially musculoskeletal trauma, but valid data regarding its effectiveness are lacking. This paper reviews the pharmacology of DMSO and reports on its effectiveness in reducing posttraumatic limb swelling and ankle joint stiffness in a rabbit hind limb model. The left and right hind limbs of the test and control animals were instrumented and fractured identically. DMSO was applied daily to the skin of only one limb in the test animals. DMSO reduced postinjury ankle stiffness in both ankles of the test rabbits by 41% but had no effect on limb swelling compared to control rabbits. Postulated mechanisms of decreased joint stiffness include oxygen-free radical scavenging and inhibition of fibroblast proliferation. PMID- 3042237 TI - Spasticity and contracture. Physiologic aspects of formation. AB - Disruption of the upper motor neuron inhibitory pathways by stroke, brain trauma, or spinal cord injury leads to muscle spasticity. Spasticity is characterized by increased muscle tone, hyperactive reflexes, and possible clonus or rigidity. The increased muscle tone may result in loss of joint motion, leading to contractures. Treatment of established contractures is difficult. Prevention of contractures by joint mobilization is emphasized as a goal in the management of patients with spasticity. PMID- 3042238 TI - [2 cases of Creutzfeldt-Jacob disease with bilateral vocal cord paralysis]. PMID- 3042239 TI - Unexpected demonstration of superior vena caval obstruction in third trimester lung imaging. AB - A large focus of liver activity was seen after the injection of Tc-99m macroaggregated albumin (MAA) in a pregnant patient thought to have pulmonary embolism (PE). The correct diagnosis of superior vena caval (SVC) obstruction led to an immediate cesarean section, CT of the chest and abdomen, open lung biopsy of a mediastinal mass, and radiation therapy for the lymphoma with pericardial invasion, all within 24 hours of presentation. The next day, an in vitro labeled Tc-99m red blood cell (RBC) angiogram was performed, which documented collateral flow from the SVC obstruction to the abdomen and filling of the right ventricle via the liver, presumably through reopened channels of the umbilical vein. PMID- 3042241 TI - Brain death: confirmation by radionuclide cerebral angiography. AB - Dynamic radionuclide cerebral angiography was performed in 14 patients with suspected brain death. In 10 of 14 patients, no intracranial arterial perfusion was demonstrable, thus confirming brain death. In four patients, faint venous activity was seen in the sagittal sinus only. All these patients also eventually died. Radionuclide cerebral angiography provides a simple and noninvasive means to confirm brain death in critically ill patients maintained on life support systems particularly when an electroencephalogram and four vessel contrast angiography may be either impractical or equivocal. PMID- 3042240 TI - Renal scintigraphic findings in a case of transplant renal vein thrombosis. AB - Renal vein thrombosis is a rare complication of renal transplantation. The scintigraphic features of a documented case of renal vein thrombosis studied with Tc-99m DTPA and the differential diagnosis relevant to these findings are discussed. PMID- 3042242 TI - Detection of abdominal hemangiomatosis during technetium-99m DTPA renal imaging. PMID- 3042243 TI - Pharmacokinetics of calcium antagonists under development. AB - Calcium antagonist drugs under clinical development are of the Type I (verapamil, diltiazem-like) and Type II (nifedipine-like) classes. Tiapamil, the only Type I drug currently available, is a high clearance, widely distributed drug which undergoes extensive presystemic elimination. Pharmacokinetically it is quite similar to verapamil; however, it does have increased biliary excretion and decreased binding to plasma proteins. Eight Type II (dihydropyridine) drugs are reviewed. Seven of these drugs (felodipine, isradipine, nicardipine, nilvadipine, nimodipine, nisoldipine and nitrendipine) are pharmacokinetically similar to nifedipine, with high clearance, extensive distribution, and significant presystemic elimination. Amlodipine has lower clearance, even greater peripheral distribution, and greatly decreased presystemic elimination. Three of the 8 dihydropyridines have been reported to have plasma protein binding greater than 90%. Unlike nifedipine, each dihydropyridine drug under development has an asymmetric centre; therefore each in fact is a racemic mixture. Human pharmacokinetic and pharmacodynamic data have not been reported for any of the racemates. Each of the drugs has been studied in patients with hepatic and renal disease. Predictably, patients with severe hepatic disease have decreased presystemic clearance and, in some cases decreased clearance after intravenous administration of the dihydropyridines, although renal failure has little influence on their pharmacokinetics. Unfortunately, disease-drug interaction studies of this class of drugs do not generally report plasma protein binding. The effect of age on the disposition of 2 of the dihydropyridines has been reported; however, only for nicardipine can a conclusion be drawn, namely that volume of distribution may increase with age and clearance may remain unchanged. A variety of potential drug-drug interactions have been evaluated, most commonly the effect of these drugs on cardiac glycoside disposition and effect, and the effect of cimetidine on the disposition of dihydropyridines. Tiapamil, like verapamil, impairs digoxin clearance significantly. Among the dihydropyridines, although minor pharmacokinetic effects have in some cases been reported, the magnitude of the interactions suggest they have limited clinical importance. From drugs currently under development, it is clear that a large number of calcium antagonists will soon be introduced into clinical use. Only 1 of the newer drugs, amlodipine, has significant pharmacokinetic differences from the agents currently in use, although possible pharmacodynamic differences among the drugs have been suggested.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3042247 TI - Ecological breastfeeding and child spacing. AB - Until the 2nd and 3rd decades of this century, breastfeeding was essential for infant survival. In that period, spacing of children was generally about 2 years. Later, improved modified cow's milk preparations became commercially available and were well tolerated by most infants. As a result, near cessation of ecological breastfeeding occurred toward the middle of the century. The decline in ecological breastfeeding was associated with early postpartum ovulation and shortened child-spacing of about 1 year. The endocrinology of breastfeeding is now known in considerable detail. Prolactin is secreted promptly in response to nipple stimulation and is a reliable marker of the endocrine alterations occurring postpartum. Success of lactation in suppression of ovulation was found to occur when infants sucked frequently and when only small amounts of selected foods were introduced gradually after the infants were about 6 months of age. PMID- 3042248 TI - Splenic infarction at low altitude in a child with hemoglobin S-C disease. AB - We describe a 15-year-old black boy with hemoglobin S-C disease living in Atlanta (altitude 1,034 ft), with no prior history of aircraft or mountain travel, who developed splenic infarction. The clinical picture was characterized by severe left upper quadrant abdominal pain, fever, splenomegaly, and hematologic and scintigraphic evidence of functional asplenia. The diagnosis was suggested by liver/spleen scintigraphy and further confirmed by ultrasonography and computerized tomography (CT) of the spleen. Treatment consisted of analgesics, intravenous fluids, and short-term antibiotic therapy. The child recovered without sequelae. PMID- 3042246 TI - Influence of the glomerular filtration rate on renal clearance of ceftazidime in cystic fibrosis. AB - The renal handling of ceftazidime was studied in 8 patients with cystic fibrosis and 10 healthy controls. The renal clearance of ceftazidime (CLRcz) was measured after an intravenous single dose and during low and high plasma concentration steady-state infusions. The glomerular filtration rate (GFR) was simultaneously estimated by inulin clearance (CL inul). The average CLRcz (mean +/- SD) was higher in cystic fibrosis patients (125 +/- 20 ml/min/1.73 m2) than in healthy controls (100 +/- 9 ml/min/1.73 m2) [p less than 0.005]. Also CL inul (mean +/- SD) was increased in cystic fibrosis patients (132 +/- 30 ml/min/1.73 m2) compared with healthy controls (103 +/- 8 ml/min/1.73 m2) [p less than 0.02]. The mean renal clearance ratios of ceftazidime to inulin were close to unity after both the single dose and low and high dose steady-state infusions both in cystic fibrosis patients and in controls. These findings suggest that the glomerular filtration rate is the principal determinant of the elimination rate of ceftazidime. However, in all cystic fibrosis patients with a CL inul exceeding 125 ml/min/1.73 m2 the clearance ratio was below unity, indicating tubular reabsorption of ceftazidime occurs in these individuals. The results demonstrate a higher but also more variable GFR in cystic fibrosis patients (74 to 174 ml/min/1.73 m2), resulting in increased and accordingly variable ability to eliminate ceftazidime in cystic fibrosis. However, these pharmacokinetic changes are not large enough to call for special dosage considerations for ceftazidime in cystic fibrosis. PMID- 3042249 TI - Group B streptococcal lymphadenitis in a child with AIDS. PMID- 3042245 TI - Ranitidine versus cimetidine. A comparison of their potential to cause clinically important drug interactions. AB - The literature on H2-antagonist drug interactions is now extensive. The whole subject is so complicated as to make things difficult for the potential prescriber. However, it is possible to reduce most of the information contained in the literature to a few simple messages. Firstly, H2-antagonists bind to cytochrome P450 and may inhibit the metabolism of drugs eliminated by the mixed function oxygenase system. In this respect, cimetidine has a marked effect which, in most studies, has reached statistical significance. Ranitidine, on the other hand, has a much weaker effect which, even if demonstrable, is statistically non significant. The potential benefit of ranitidine, however, has to be weighed against the relative costs of the 2 drugs. Secondly, H2-antagonists inhibit gastric acid production and may alter the rate of gastric emptying, and hence the rate of drug absorption. They may also have some effect on portal vein and hepatic artery flow. However, these effects are small and probably not clinically relevant. Thirdly, pharmacodynamic effects of H2-antagonist-drug interactions are difficult to demonstrate in planned studies, and although they are reported from time to time, adverse reactions of consequence are relatively uncommon. Fourthly, the prescriber needs to be aware that cimetidine may produce higher plasma concentrations of some drugs which have a fairly narrow therapeutic range, and this may be clinically important. Examples of drugs for which it may be undesirable to inadvertently increase plasma concentrations include warfarin, theophylline and phenytoin. Finally, for most drugs metabolised by the liver, the risk of an important interaction is small. However, if such an interaction is noted it may be helpful to refer to the other reported cases, and a number of references are included here. PMID- 3042244 TI - Clinical pharmacokinetics of doxorubicin. AB - Doxorubicin (adriamycin) has a very wide antitumour spectrum, compared with other anticancer drugs; however, except for Hodgkin's disease, it is not associated with curative chemotherapy. Doxorubicin has been in clinical use for more than 2 decades, and only recently has it been recognised that the cytotoxic effect is produced at the cellular level by multiple mechanisms which have not yet been conclusively identified. Key factors are a combination of doxorubicin-induced free radical formation due to metabolic activation, deleterious actions at the level of the membrane, and drug-intercalation into DNA. Multiple aspects of the clinical pharmacokinetics of this drug have been described. Wide interpatient variations in plasma pharmacokinetics have been noted, but without firm relation to clinical outcome. An apparent volume of distribution of approximately 25 L/kg points to extensive uptake by tissues. Up to several weeks after administration, significant concentrations of doxorubicin have been found in haematopoietic cells and in several other tissues. The maximum cellular doxorubicin concentrations reached in vivo remain significantly below those at which all clonogenic leukaemic cells are killed in vitro. Doxorubicin has been administered as frequent (weekly) low doses, single high doses, and as a continuous infusion. The optimal schedule with respect to tumour cytotoxicity and dose-limiting side effects such as myelosuppression or cardiotoxicity, has never been investigated in a prospective, randomised manner. Clinical trials large enough to study optimal, and possibly individualised, doxorubicin chemotherapy need to be performed. This review summarises pharmacological and pharmacodynamic data of doxorubicin, and discusses these in relation to possible improvement of its therapeutic index. Furthermore, drug interactions, dose-response relationships, mechanisms of action, multidrug resistance, and treatment scheduling are discussed in the perspective of the development of novel treatment strategies. PMID- 3042250 TI - Renin-angiotensin system, blood pressure homeostasis and renal function in galactosamine-induced fulminant hepatic failure in the guinea pig. AB - Arterial blood pressure, renal function and plasma concentrations of renin and renin substrate (angiotensinogen) were investigated in guinea pigs subjected to galactosamine-induced (1 g/kg i.v.) liver cell necrosis. Blood pressure declined continuously by 50% during a follow-up period of 72 h which was associated with a decrease in diuresis and natriuresis to 36 and 31%, respectively. Simultaneously, plasma renin concentration increased 30-fold indicating marked reduction of renal perfusion, while plasma renin substrate concentration fell to 6% of the baseline level. There was microscopic evidence of oligemic circulatory renal damage characterized by acute proximal tubular necrosis with concomitant tubular dilatation. Short-term infusion of homologous renin substrate-enriched plasma, derived from nephrectomized animals, was followed by marked increase in mean arterial blood pressure from 34 +/- 9 to 77 +/- 7 mm Hg accompanied by marked diuresis and natriuresis. Renin substrate depletion following galactosamine induced fulminant liver failure may represent impaired hepatic biosynthesis as well as increased renin substrate consumption due to excessive renin secretion. Angiotensinogen repletion has a beneficial effect on both renal function and blood pressure probably due to marked generation of the potent vasoconstrictor angiotensin II which consequently inhibits renin secretion. These observations strongly support the suggestion that the renin-angiotensin system is of major importance to cardiovascular homeostasis in acute liver failure. PMID- 3042251 TI - The natural history of brittle diabetes. AB - The natural history of brittle diabetes is unknown. We have followed up 13 patients with disabling brittle diabetes unresponsive to continuous subcutaneous insulin infusion (CSII) for 3-6 yr. All were young, C-peptide deficient females. One patient has died (of hypoglycaemia). In the others, disruption of life has generally lessened, but only one patient is currently considered metabolically stable. Insulin treatment regimens have included long-term intravenous insulin infusions and intraperitoneal insulin, but all but four have now reverted to subcutaneous injections. Eleven patients intermittently required greater than 200 U/day of insulin and two have needed greater than 1,000 U/day. Insulin dosages have fallen significantly during follow-up (from 6.8 +/- 3.1 to 1.4 +/- 0.3 U/kg/day). Diabetic complications, initially present in only 2 cases (1 cataract, 1 proliferative retinopathy), have now developed in 5 others (2 background retinopathy, 1 proliferative retinopathy, 1 mixed peripheral neuropathy and 1 intermittent proteinuria). Psychosocial disturbance and non-compliance were common. We conclude that brittleness generally seems to improve, which probably explains the scarcity of older brittle patients. However, these patients are at considerable risk from diabetes, its complications and its treatment. PMID- 3042254 TI - Evidence for a critical role of diet in the development of insulin-dependent diabetes mellitus. AB - The role of diet in human insulin-dependent diabetes mellitus (IDDM) has not been properly examined, mainly due to a lack of reliable markers to identify prospective diabetics and the difficulties in obtaining accurate and representative dietary information. Nonetheless, there is some circumstantial evidence suggesting a role for diet in human IDDM. The validity of this relationship in humans must await a sufficiently large, well designed prospective study or the discovery of better markers for diabetes predisposition. The recent availability of the spontaneously diabetic BB rat has permitted prospective studies under controlled conditions which indicate diet, particularly dietary proteins, such as wheat gluten or cow's milk proteins may be prerequisites for maximum expression of the insulin-dependent syndrome in these animals. The early suckling and/or post-weaning period appears to be important and may be the crucial time when these proteins or portions of them pass the gastrointestinal barrier and initiate a process, possibly immunological, which results in destruction of the beta cells of the pancreas. PMID- 3042252 TI - Soluble and Lente human insulin mixtures in normal man. AB - The interaction between soluble (Actrapid HM) and Lente (Monotard HM) human insulin preparations was examined in normal subjects. Incremental plasma insulin levels were determined following the subcutaneous administration of 6 U of soluble insulin admixed with 0.14 ml soluble insulin diluting medium, or 0.14 ml Lente diluting medium and injected either immediately or 5 min after preparation. For comparison separate but simultaneous injections of soluble (6 U) and Lente insulin (14 U) were administered subcutaneously. All injection volumes were identical. The incremental insulin levels were significantly greater between 60 and 90 min (p less than 0.05-0.01) following the administration of soluble insulin admixed with its own medium compared to the soluble insulin/Lente medium admixtures injected immediately and 5 min after preparation. There was no difference, however, in the plasma insulin profiles between the two soluble insulin/Lente medium schedules. The separate simultaneous administration of soluble and Lente insulin resulted in significantly higher plasma insulin levels at 30 and 90 min (p less than 0.05) when compared to the admixture injected immediately after preparation and from 30-120 min (p less than 0.05-0.01) compared to the admixture injected after 5 min. The two soluble/Lente insulin admixtures achieved similar plasma insulin profiles. Therefore when soluble human insulin is admixed with Lente human insulin and administered by subcutaneous injection immediately after admixing there is a significant reduction in the plasma insulin levels during the first 90 min in contrast to when the two preparations are given simultaneously by separate injection. Delaying the injection of the admixture for 5 min results in significantly lower insulin levels up to 120 min. These differences are reflected in the hypoglycaemic responses observed. PMID- 3042253 TI - Clinical response to feeding a high polyunsaturated fat diet in normal and diabetic rats. AB - Dietary fat composition can influence membrane composition in a variety of cell types. Changes in dietary fat can alter the structure and function of lymphocyte membranes, liver plasma membranes, brain synaptosomes and cardiac mitochondria. In diabetics, uptake of glucose is enhanced due to a greater passive permeability of the intestine to glucose. This enhanced absorption from the intestine can be partially corrected by insulin administration and islet cell transplantation. In clinical practice, however, a more commonly used therapeutic modality for diabetics is diet modification. In this study, when diabetic rats were fed a high fat diet, high or low saturated to polyunsaturated fatty acid, there was an improvement in the plasma and urine glucose. This improved clinical response was also observed in the glycosylated hemoglobin response in the animals fed the high polyunsaturated to saturated fatty acid diet. The clinical improvement observed after feeding with these diets, however, was not seen during the oral and intravenous glucose tolerance tests. This study shows that dietary alteration can improve the clinical response in diabetic animals. PMID- 3042255 TI - Proinsulin, insulin, and C-peptide in cystic fibrosis after an oral glucose tolerance test. AB - The beta-cell response to an oral glucose load was studied in 22 patients with cystic fibrosis (CF) by means of insulin, C-peptide and proinsulin, and the results compared with those from 20 healthy sex and age matched controls. According to WHO criteria two had diabetes mellitus, eight had impaired glucose tolerance and twelve had normal glucose tolerance. All patients showed lower insulin and C-peptide levels than the controls at 30 minutes. However the insulin and C-peptide responses were sustained so that the areas under the curves were comparable between controls, CF patients with normal glucose tolerance, and CF patients with impaired glucose tolerance. By contrast, the area under the proinsulin curve was significantly higher in the CF patients with impaired glucose tolerance compared with both controls (p less than 0.05) and with the CF normal glucose tolerance group (p less than 0.02). In the CF patients with impaired glucose tolerance the proinsulin level was significantly elevated at 120 minutes (median 50 pmol/l) and at 180 minutes (29 pmol/l) as compared with the controls (24 and 19 pmol/l p less than 0.01) and with the CF patients with normal glucose tolerance (21 and 16 pmol/l p less than 0.01). These data confirm that impaired glucose tolerance and diabetes is frequent in cystic fibrosis. The elevated proinsulin immunoreactive material in CF patients with impaired glucose tolerance may partly compensate for the relative insufficient insulin response found in these patients. PMID- 3042256 TI - Basolateral binding and uptake of 125-I-insulin in proximal tubular cells after peritubular extraction in the avian kidney. AB - Several studies have suggested the existence of a renal peritubular extraction of insulin. So far, however, no conclusive evidence, as to whether insulin enters the proximal tubular cell following peritubular extraction, has been presented. In the present study we injected iodine labelled porcine insulin into the renal portal system on hens prepared according to a modified Sperber technique and followed cellular handling of the extracted 125-I-insulin using electron microscope autoradiography at 1 and 7 min after injection. The results showed that at 1 min after injection, peritubular extraction of 125-I-insulin accounted for as much as 30% of total proximal tubular accumulation of grains. Of these grains about 40% were located over basal vesicles, lysosomers or other cell organelles with the remaining 60% located in the intercellular space, probably bound to the basolateral membrane. At 7 min after injection the distribution of grains subsequent to peritubular extraction of 125-I-insulin was unchanged. In contrast, very few grains were located over distal tubules at 1 or 7 min. Thus, the results demonstrate a sizable peritubular extraction of 125-I-insulin in the avian kidney with subsequent uptake into the proximal tubular cells of about one third of the extracted insulin. PMID- 3042257 TI - [Rapid magnetic resonance tomography sequences--theoretical principles and clinical imaging characteristics]. AB - This paper is concerned with fast MR-imaging, realised by several "fast sequences", introduced during the last few years into clinical routine diagnostics. Two of the most important fast sequences are FLASH and FISP--their physical and technical working as well as their relevant clinical imaging properties are described. PMID- 3042258 TI - [Diagnosis of a perforating aortic aneurysm as a late complication following enlarging aortoplasty using central venous digital subtraction angiography]. AB - 11 years after patch aortoplasty due to coarctation of the aorta a 33 year-old patient experienced two spontaneous haemoptyses. A centralvenous digital subtraction angiography demonstrated a large aneurysm of the proximal descending aorta penetrating the left upper lobe of the lung. The successful emergency surgical treatment consisted of implantation of a dacron-prosthesis. PMID- 3042259 TI - [Ultrasound findings in patients with AIDS]. AB - The ultrasonographic findings of 43 patients with AIDS and ARC were analyzed. In 63% an enlarged liver, in 66% an enlarged spleen, partially with focal lesions, and in 21% enlarged abdominal lymph nodes were diagnosed. The typical parenchymal lesions of the kidney (focal segmental glomerulosclerosis) were not observed. Abdominal ultrasound is the first diagnostic procedure to perform in patients with AIDS and ARC with the suspicion of abdominal pathology. With additional thin needle puncture of the lesions a histological verification of the pathologic findings is possible. PMID- 3042260 TI - [Duplex sonographic findings following kidney transplantation-- possible value in the diagnosis of graft rejection]. AB - 59 prospective duplex ultrasound studies have been performed on 30 patients after renal transplant to assess the feasibility of detection of acute rejection. The results were compared with clinical, biochemical and histopathologic findings. Real time ultrasound was not a reliable method in predicting acute rejection. Doppler sonography, however, characteristically showed a diminished diastolic blood flow in all cases of acute rejection. Acute tubular necrosis and cyclosporine toxicity were not provable, neither by real time ultrasound nor by doppler sonography. Duplex ultrasound combines the advantages of both methods in detecting perirenal fluid collections, hydronephrosis and acute transplant rejection. PMID- 3042261 TI - The expression of S-100 protein and neuron-specific enolase in meningiomas. AB - The distribution of S-100 protein and neuron-specific enolase (NSE) was examined in 19 meningiomas using the peroxidase-antiperoxidase (PAP) immunohistochemical technique. Positive cytoplasmic staining for NSE was observed in 16 tumors, and comparable staining for S-100 protein was observed in eight tumors. The finding of S-100 protein and NSE immunoreactivity in fibroblastic, meningiotheliomatous, and transitional meningiomas raises the possibility that these morphologically distinct neoplasms derive from a common pluripotential cell capable of differentiation along diverse paths. PMID- 3042263 TI - Circulating immune complexes in malignant melanoma. PMID- 3042262 TI - The metastatic phenotype. PMID- 3042264 TI - Detection of malignant melanoma with monoclonal antibodies. AB - Eleven murine monoclonal antibodies (MoAbs) were isolated that defined unique membrane antigens expressed on human melanoma cells but not detectable on human lymphoid cells by radioimmunometric assays. Five of these MoAbs each identified a separate melanoma cell surface antigen as shown by distinctly different in vitro MoAb binding patterns to a diverse panel of tumor cell lines. One of these 5 monoclonals, MoAb 34.1, reacted specifically with 9/11 melanoma lines and 0/28 other human tumor or lymphoid cell lines. The other 4 MoAbs reacted strongly with melanomas, but also bound to 1 or more non-melanoma lines. The remaining 6 MoAbs defined three distinct regions of a single melanoma cell membrane protein with a molecular weight of 125 kiloDaltons (kD) as shown by antibody crossblocking and gel electrophoresis. A sensitive radioimmunoassay developed with MoAbs to 2 epitopes of this 125 kD protein detected up to 500-fold higher levels of this antigen in extracts of melanoma cells compared to autologous lymphoid cells. The 125 kD antigen also was detected by indirect immunoperoxidase assays with the MoAbs on biopsied tumors in histologic tissue sections of 5/11 metastatic melanomas and 1/11 carcinomas but was found on some normal endothelium and smooth muscle. Another monoclonal, MoAb 705, reacted more broadly with tumor cells in 10/14 biopsied melanomas and 10/11 carcinomas, but also was reactive with basal epidermis and normal fibroblasts. By contrast, MoAb 34.1 bound specifically to tumor cells of 7/11 biopsied metastatic melanomas, but bound 0/10 carcinomas and few normal tissues except for some macrophages. Thus, MoAb 34.1 was the most specific diagnostic reagent for immunohistologic detection of melanoma. The 250 kD antigen defined by MoAb 34.1 is similar to a high molecular weight proteoglycan reported to be an excellent tumor marker for human melanomas. The results of these studies show that murine monoclonal antibodies can be used as sensitive reagents for radioimmunoassays and immunohistology of malignant melanoma. PMID- 3042266 TI - Medically unnecessary denials in Medicare reimbursement. PMID- 3042265 TI - Medicare changes that will affect your practice. PMID- 3042267 TI - Clinical use of systemic antifungal agents. AB - The chemistry, pharmacology, pharmacokinetics, clinical uses, adverse effects, and drug interactions of amphotericin B, flucytosine, ketoconazole, and miconazole are reviewed. Amphotericin B, a heptaene compound with poor water solubility, disrupts the fungal cell wall by binding to ergosterol. Ketoconazole and miconazole, imidazole derivatives, are poorly water soluble and inhibit the synthesis of ergosterol. Flucytosine is a readily water-soluble, fluorinated pyrimidine agent that may be metabolized to fluorouracil. The pharmacokinetics of amphotericin B is unique and has not yet been clearly defined. After oral administration, absorption of flucytosine from the gastrointestinal tract is rapid and nearly complete. In adults, oral administration of ketoconazole produces peak concentrations of drug one to two hours after the dose. Miconazole is administered only intravenously and distributes well into most tissues. Amphotericin B remains the drug of choice for most systemic mycoses. Dosing of amphotericin B is often empiric and patient specific. Flucytosine is rarely used alone; the combination of flucytosine and amphotericin B exerts synergistic killing of many fungi. Ketoconazole is effective for treating many chronic fungal infections. Miconazole is seldom used because of the availability of agents that are equally effective, less toxic, or both. Nephrotoxicity can occur with amphotericin B therapy, while flucytosine is associated with gastrointestinal and hematologic toxicities. Ketoconazole is much less toxic than any of the other agents, while miconazole has a high incidence of adverse effects. In addition to the need for more effective and less toxic agents, research is needed to clearly define the pharmacokinetics and pharmacodynamics of currently available antifungal drugs. PMID- 3042268 TI - Effect of pentoxifylline and its metabolites on three theophylline assays. PMID- 3042269 TI - Accuracy and time requirements for use of three rapid theophylline assay methods. PMID- 3042270 TI - Thrombolysis for acute myocardial infarction: does a greater incidence of reperfusion justify preferential use of an agent? PMID- 3042271 TI - Thrombolysis for acute myocardial infarction: is proof of a reduction in mortality critical to drug selection? PMID- 3042272 TI - Prostacyclin and regional coronary blood flow in experimental myocardial infarction. AB - Infusion of prostacyclin (PGI2) has been reported to affect infarct size and myocardial blood flow favourably in various animal models of myocardial ischaemia. Recent data suggest that a similar effect of PGI2 may occur also in humans with acute myocardial infarction. We addressed the hypothesis that PGI2 redistributes myocardial blood flow following coronary ligation, and that this effect favours perfusion of myocardium at risk and thereby limits infarct size. Following ligation of a distal branch of the left coronary artery in anaesthetized dogs, PGI2 (2-4 ng/kg/min) was infused for 72 h. Regional myocardial blood flow was assessed immediately after the coronary ligation and at the end of the drug infusion, by injection of 57Co- and 113Sn-labelled microspheres, respectively. Coronary ligation reduced regional coronary blood flow by 40-70%. During the subsequent 72 h the blood flow increased, being at the end of the period 50-70% of the flow in the non-ischaemic myocardium. PGI2 did not affect the spontaneous improvement of regional myocardial blood flow, as assessed at the end of the infusion. PGI2 also failed to affect infarct size, either when expressed in relation to total left ventricular mass, or in relation to area at risk. We conclude that PGI2, when infused immediately after coronary ligation in dogs in a clinically relevant dose, neither affects regional myocardial blood flow in the ischaemic regions, nor the size of the myocardial infarction. PMID- 3042274 TI - People and garbage are not the same: issues in contracting for public mental health services. PMID- 3042273 TI - Cardiovascular effects of positive end-expiratory pressure during acute left ventricular failure in dogs. AB - Haemodynamic and metabolic effects of ventilation with positive end-expiratory pressure (PEEP) were studied in closed-chest dogs anaesthetized with sodium pentobarbital during normal cardiac function and during acute left ventricular (LV) failure. LV failure was induced by embolizing the left coronary bed with 50 micron plastic microspheres causing a marked depression of LV function. End expiratory pressure was set to 0, 5, 10 and 15 cm H2O both before and after coronary embolization. During normal cardiac function PEEP above 5 cm H2O depressed cardiac output (CO) significantly. However, following coronary embolization after which LV function was seriously impaired, CO was maintained as PEEP was applied. This is attributed to reduced sensitivity to the LV pre-load reductions induced by PEEP during LV failure. PEEP reduced MBF both during normal and impaired LV function. This did not result in ischaemic myocardial metabolism assessed by lactate extraction either in normal hearts or following coronary embolization. The reduced MBF was, however, associated with reduced MVO2 both during normal cardiac function and during LV failure. It is suggested that the reduced MBF is mainly due to reduced myocardial oxygen demand probably caused by reduced LV wall tension during PEEP ventilation. PMID- 3042276 TI - Summary of presidency. PMID- 3042275 TI - Buprenorphine. PMID- 3042277 TI - Effects of flunixin meglumine in dogs following experimentally induced endotoxemia. AB - Twelve dogs were randomly divided into three groups. Group 1 dogs were given Escherichia coli endotoxin and then treated with flunixin meglumine. Group 2 dogs were given endotoxin as group 1, but untreated. Group 3 dogs were given flunixin meglumine alone. The dogs were monitored clinically and urine and serum samples were collected at regular intervals for 72 hours. All surviving dogs were humanely killed after 72 hours and examined for gross and histologic lesions. Group 1 dogs all survived 72 hours, but showed prerenal azotemia, hepatocellular damage, hemorrhagic enteritis, and numerous gastric ulcerations. Three of the four dogs in group 2 died before 72 hours. Group 2 dogs showed many of the same chemical and hemodynamic changes as group 1. They had severe hemorrhage into the intestinal lumen; however, there were no gastric ulcerations. Group 3 dogs all survived and showed little physical or hematologic change. The study suggested the following: 1) flunixin meglumine was an effective drug in ameliorating the fatal effects of canine endotoxemia, 2) the effects of endotoxin in combination with flunixin meglumine, at 1.1 mg/kg body weight, caused gastric ulcerations, and 3) in normal dogs flunixin meglumine at 1.1 mg/kg body weight did not cause severe side effects or gross lesions. PMID- 3042278 TI - Neoplastic angioendotheliomatosis in a dog: an angiotropic lymphoma. AB - Neoplastic angioendotheliomatosis was diagnosed in a nine-year-old, intact male German Shorthaired Pointer exhibiting progressive neurological signs over a six month period. At necropsy, there was multifocal asymmetric, hemorrhagic necrosis within the cerebral hemispheres, centrum semiovale, caudate nucleus, and internal capsule. Histologically, there was extensive intravascular proliferation of pleomorphic mononuclear cells within the brain and multiple parenchymatous organs. Transmission electron microscopy demonstrated the absence of intercellular junctions between neoplastic cells. These cells were not attached to the vascular lining endothelium; Weibel-Palade bodies and a basement membrane were lacking. By indirect immunofluorescence, positive cytoplasmic staining of intravascular neoplastic cells for IgG was demonstrated. Peroxidase antiperoxidase technique for Factor VIII related antigen was negative. As in man, this rare neoplastic disorder appears to be a lymphoma, apparently of the B cell line. PMID- 3042279 TI - Transscleral YAG laser photocoagulation for uncontrollable glaucoma in corneal patients. AB - Control of intraocular pressure (IOP) in patients with severe alkaline burns is one of the most difficult problems facing the corneal specialist. Currently, when medical therapy cannot control intraocular pressure, the usual procedure of choice is cyclocryotherapy. This procedure, however, can be complicated by phthisis, retinal detachment, or macular edema. We have used transscleral YAG laser cyclophotocoagulation (TSYLC) to control the IOP in a patient with severe glaucoma after an alkali burn. This patient had previously had an unsuccessful cyclocryotherapy. After the TSYLC procedure, he suffered no complications and his IOP was normal. Our experience with this patient indicated that the TSYLC procedure might be an effective alternate to cyclocryotherapy, especially in corneal patients with uncontrollable inflammatory glaucoma. PMID- 3042280 TI - Modified relaxing incision technique for postkeratoplasty astigmatism. AB - A modified relaxing incision technique for postkeratoplasty astigmatism is described in this article. Following the initial, standard relaxing incision techniques, a planned spreading and recutting of the initial relaxing incision was performed at 1-3-week intervals following the initial surgery. The wound may be deepened and lengthened if more effect is needed. The number of spreading and recutting procedures is also dependent on the effect required. This additional procedure may be performed for late regression of the desired effect. The endpoint for the procedure is corneal astigmatism that will allow either spectacle or contact lens correction, depending on the patient's visual needs. Corrections of greater than 10 D of astigmatism are possible. The technique allows for evaluation of the corneal astigmatism with no sutures in place. No complications of the recutting and spreading procedure have been noted. There have been no microperforations or macroperforations requiring suturing, no infections, and no graft rejections following the procedures. Seven cases using the modified relaxing incision technique are described. PMID- 3042281 TI - Surgical correction of postoperative astigmatism. AB - The photokeratoscope has increased the understanding of the aspheric nature of the cornea as well as a better understanding of normal corneal topography. This has significantly affected the development of newer and more predictable models of surgical astigmatic correction. Relaxing incisions effectively flatten the steeper meridian an equivalent amount as they steepen the flatter meridian. The net change in spherical equivalent is therefore negligible. Poor predictability is the major limitation of relaxing incisions. Wedge resection can correct large degrees of postkeratoplasty astigmatism. Resection of 0.10 mm of tissue results in approximately 2 diopters of astigmatic correction. Prolonged postoperative rehabilitation and induced irregular astigmatism are limitations of the procedure. Transverse incisions flatten the steeper meridian an equivalent amount as they steepen the flatter meridian. Semiradial incisions result in two times the amount of flattening in the meridian of the incision compared to the meridian 90 degrees away. Combination of transverse incisions with semiradial incisions describes the trapezoidal astigmatic keratotomy. This procedure may correct from 5.5 to 11.0 diopters depending upon the age of the patient. The use of the surgical keratometer is helpful in assessing a proper endpoint during surgical correction of astigmatism. PMID- 3042282 TI - Donor failure after corneal transplantation surgery. AB - Donor failure refers to graft edema present within the first 24 h after penetrating keratoplasty that persists in spite of maximal medical therapy. We reviewed the case histories of 1,351 penetrating keratoplasties. Of these, 17 (1.2%) were considered donor failures. Five cases were histopathologically consistent with Fuchs' dystrophy. Seven cases demonstrated mechanical stripping of the endothelium. Five cases, all from the same eye bank, revealed an absence of posterior stromal keratocytes. Storage medium at the eye bank of origin was found to have an alkaline pH. Careful preoperative evaluation of donor corneas is emphasized, but if an apparent "epidemic" of donor failure occurs, a thorough investigation of the eye bank methodology should be initiated. PMID- 3042283 TI - Growth factors and corneal epithelium. AB - Regeneration of corneal epithelium following injury is essential for visual rehabilitation. A limited number of approaches are available for treating patients who fail to heal epithelial injuries adequately. The presence of specific receptors for epidermal growth factor (EGF) on epithelial cells suggests that this potent mitogen may play a role in normal epithelial wound healing. Topical application of biosynthetic human EGF significantly accelerated epithelial regeneration in primates following epikeratophakia surgery. Epidermal growth factor alone and with fibronectin accelerated epithelial regeneration of rabbits following mild alkali burns. Since prolonged exposure of cells to EGF is necessary to induce mitosis, the dynamics of EGF in the eye and with various lenses was studied. When applied in methylcellulose-based eye drops, 90% of the EGF was lost from tear film within 10 min, while a small amount (10%) remained associated with conjuctival tissue. Soft contact lenses or epikeratophakia lenticles took up substantial amounts of EGF (50 micrograms) and released 85% of the EGF within 24 h, with a half-life of 4 h in vitro. Epidermal growth factor did not diffuse through corneas or lenticles and did not promote epithelial downgrowth along sutures in primate corneas. These results suggest that biosynthetic growth factors may be useful in the treatment of some epithelial injuries. PMID- 3042284 TI - Prehospital thrombolytic treatment of acute myocardial infarction with anisoylated plasminogen streptokinase activator complex. AB - In cooperation with a group of general practitioners (GP), we investigated the possible risk and benefit of prehospital initiation of thrombolytic therapy in acute myocardial infarction (AMI) with anisoylated plasminogen streptokinase activator complex (APSAC) at the patient's home. During a 14-month period, 58 patients with suspected AMI were evaluated by their GP using a protocol with strict inclusion and exclusion criteria. The GP alerted a special mobile intervention team which administered APSAC at home in 13 of the 19 patients. Coronary reperfusion was achieved in ten of these 13 patients. Apart from short and easily treated episodes of bradycardia and/or hypotension after the injection of the thrombolytic drug in four of 13 patients, no major adverse events were noted in the early treatment period. The estimated time gain by treating the patient at home instead of starting the treatment in the coronary care unit was 46 +/- 14 min. Therefore, at-home initiation of thrombolytic treatment seems feasible, fast, and safe. PMID- 3042285 TI - Prospective study of catheter-related infection during prolonged arterial catheterization. AB - Ninety-six arterial catheters from 75 different anatomical sites in 56 surgical ICU patients were studied prospectively to determine the rate of catheter-related infection associated with prolonged arterial catheterization (defined as greater than 96 h). Every 96 h, all catheters were semiquantitatively (SQ) cultured and the percutaneous entry site was swab cultured. Sites were used indefinitely by exchanging the catheters over a guide-wire every 96 h as long as arterial monitoring was necessary and SQ cultures remained negative (less than or equal to 15 colonies). No sites used less than 96 h developed skin colonization, while 14/51 (27%) sites used greater than 96 h developed positive swab cultures. No SQ cultures were positive in sites with negative swab cultures (p less than .001). Catheter-related infection (a positive SQ culture) developed in 4/42 (9.5%) radial or femoral sites compared to 4/9 (44%) axillary sites used greater than 96 h (p less than .01). It is concluded that arterial catheter-related infection develops in less than 10% of radial or femoral sites used greater than 96 h, and 90% of radial and femoral sites may be used safely for prolonged periods if skin colonization at the percutaneous sites is controlled and SQ catheter cultures remain negative. Skin site swab cultures may be useful for determining when arterial catheters should be removed and SQ cultured. PMID- 3042287 TI - Some infant ventilators do not limit peak inspiratory pressure reliably during active expiration. AB - In order to minimize barotrauma in newborn infants with respiratory failure, peak inspiratory pressures should not exceed those required for adequate gas exchange. We examined whether four commonly used pressure-limited, constant flow ventilators limit pressure reliably during simulated active expiration against the inspiratory stroke of the ventilator. Three machines of each type were tested at 13 different expiratory flow rates (2 to 14 L/min). Flow-dependent pressure overshoot above a dialed pressure limit of 20 cm H2O was observed in all machines. However, the magnitude differed significantly between ventilators from different manufacturers (p = .0009). Pressure overshoot above 20 cm H2O was consistently lowest in the Healthdyne (0.8 cm H2O at 2 L/min, 3.6 cm H2O at 14 L/min) and highest in the Bourns BP200 (3.0 cm H2O at 2 L/min, 15.4 cm H2O at 14 L/min). We conclude that peak inspiratory pressure overshoots on pressure-limited ventilators occur during asynchronous expiration. This shortcoming may contribute to barotrauma in newborn infants who "fight" positive-pressure ventilation. PMID- 3042286 TI - Effect of scavengers of oxygen-derived free radicals on mortality in endotoxin challenged mice. AB - Oxygen-derived free radicals have been implicated as mediators of cellular injury in several model systems. Recently, a role for free radicals has been proposed in the mortality associated with Gram-negative bacterial sepsis. To determine if pretreatment with free radical scavengers can prevent endotoxin-induced mortality, mice rendered sensitive to endotoxin with actinomycin D were treated with either superoxide dismutase (SOD), N-acetylcysteine (NAC) or saline and were then challenged with a dose of endotoxin calculated to cause a mortality of greater than 80%. Mortality was assessed at 12-h intervals after challenge. Increased survival was seen in the SOD-treated group compared to the control group (p less than or equal to .05). In contrast, survival in mice treated with NAC, another potential scavenger, was not significantly different from the control group. These results support the hypothesis that superoxide and hydroxyl radicals contribute to mortality in Gram-negative bacterial sepsis. PMID- 3042288 TI - Gastric ulcers: role of oxygen radicals. PMID- 3042289 TI - In vivo and in vitro deposition of immunoglobulin aggregates in the mouse eye. AB - The possible role of specific mechanisms involved in the adherence process of immune aggregates to tissue components of the mouse eye was investigated in an experimental animal model. Passive intravenous administration of immunoglobulin aggregates to mice, resulted in the localisation of these aggregates in various organs, including the eye. In the eye a strong localisation was seen in the episcleral capillary plexus, whereas only a weak deposition was seen in the iris, ciliary body and choroid. No deposits were seen in the retina. To investigate the role of specific receptors for immune complexes in the eye, in vitro experiments were performed, whereby immunoglobulin aggregates were layered on cryostat sections of the mouse eye. These in vitro studies also showed an adherence of immune aggregates to the episcleral capillary region, but furthermore a deposition on mast cells scattered throughout the extra-ocular muscles. The in vitro binding of immunoglobulin aggregates to the episcleral capillary plexus could be inhibited by high concentrations of Fc fragments and monomeric IgG, but not with Fab fragments or 0.5 M NaCl. The in vitro adherence of aggregates to mast cells could not be blocked by the inhibitors employed in our study and could therefore be distinguished from the interaction of aggregates with the episcleral capillary plexus. These results indicate that the ocular deposition of immunoglobulin aggregates in the episcleral capillary plexus of the mouse eye is (immune) specific and mediated by Fc receptors. PMID- 3042290 TI - Recognition and management of inflammatory bowel disease in children and adolescents. AB - Until such time as an etiology and specific therapy can be established, it is critical for the pediatrician to recognize the early clinical manifestations of IBD and to initiate appropriate therapy. Most affected children require consultative care with a gastroenterologist comfortable with the unique requirements of the child with IBD. It is our hope that early diagnosis and treatment will not only reduce the frequency and severity of complications, but also allow minimal interference in the child's quality of life. PMID- 3042291 TI - Interventional computed tomography. AB - Due to the development and refinement of computed tomography (CT), sonography, and interventional techniques, the field of interventional radiology has seen tremendous growth in recent years. In particular, the precise anatomic detail provided by CT and sonography has allowed percutaneous biopsies and abscess drainages to be performed safely and effectively. Percutaneous biopsies are now becoming the most common interventional radiographic procedures in many institutions. The usual indications for a biopsy are to determine the etiology of a mass, neoplasm, or inflammation, and to determine whether masses in known oncologic patients represent scarring or residual viable tumor. Accuracy rates for most percutaneous CT-directed biopsies are well over 90%, and the complication rate is very low. CT-directed percutaneous abscess drainages are also safe and effective and, in most cases, will be preferable to surgical drainage. The initial indications for percutaneous drainage (single, unilocular fluid collections) have been greatly expanded to include multiloculated collections, interloop abscesses, periappendiceal abscesses, and even percutaneous cholecystotomies. Biopsy and drainage procedures, together with their accuracy rates, indications and complications, are reviewed in this monograph. PMID- 3042292 TI - Successful hepatic preservation using pulsatile perfusion and allopurinol. PMID- 3042293 TI - Acute liver failure after subtotal hepatectomy in the rat: failure of testosterone pretreatment and hepatocyte transplantation to improve survival. PMID- 3042294 TI - The use of positive end expiratory pressure to alter systemic-pulmonary shunt flow using conventional and high frequency jet ventilators. PMID- 3042296 TI - [Development and clinical evaluation of a new denture adhesive]. PMID- 3042297 TI - [Experimental study on the comparison of iliac and mandibular grafts in the reconstruction of mandibular defects]. PMID- 3042298 TI - [Examination of cervical segments of the carotid and vertebral arteries by color coded continuous real-time Doppler echoflowmeter]. PMID- 3042295 TI - Menstrually related mood disorder in developmentally disabled adolescents: review and current status. PMID- 3042299 TI - Practical considerations for the selection and use of optical filters in flow cytometry. AB - The selection of proper optical filters for various excitation and emission requirements is critical in flow cytometry. Problems which arise in the selection and utilization of optical filters, and the solutions to these problems, are the subject of this article. PMID- 3042300 TI - Identification of cell subpopulations by dual-color surface immunofluorescence using biotinylated and unlabeled monoclonal antibodies. AB - A new method of dual-color immunofluorescence is presented for analysis of surface antigen distribution among heterogeneous cell suspensions. It involves flow cytometric analysis of cells stained with a biotinylated first monoclonal antibody and/or with an unlabeled second monoclonal antibody. After addition of streptavidin-phycoerythrin and/or fluoresceinated goat antimouse immunoglobulin antibody, single-cell fluorescence intensities are measured and biparametric graphic representations are obtained, allowing one to determine the percentage of cells stained by each of the monoclonal antibodies or both. The validity of the method was assessed on human peripheral blood mononuclear cells by using three sets of two monoclonal antibodies: CD8 and CD5, CD3 and CD4, CD11 and HLA-DR. The results showed that dual staining did not induce significant quenching or competition between pairs of antibodies. The procedure is simple and sensitive. It requires only minute amounts of monoclonal antibodies. It is readily applicable to the screening of hybridoma supernatants and to the characterization of new antibodies to cell surface antigens with respect to well-defined markers. PMID- 3042301 TI - The Whitehead hemorrhoidectomy. An unjustly maligned procedure. AB - The Whitehead technique of hemorrhoidectomy has developed a reputation as an undesirable procedure since its description in 1882. The chief criticisms have been considerable blood loss, disturbance of continence, formation of an ectropion, and poor healing of the mucocutaneous junction followed by stricture formation. Five hundred fifty-six patients underwent a modified Whitehead hemorrhoidectomy, performed by one author (C.E.C.), between 1963 and 1983. Seventy-two of these patients had unclaimed follow-up letters, leaving 484 patients available for review. Four hundred forty of these patients were followed for over three years. Postoperative complications included fistula or abscess in 1.1 percent, flap loss in 6.9 percent, and a nonhealing wound in one patient. There were no recurrences and there was no ectropion formation, or "Whitehead deformity." Mortality was zero and total morbidity, including 7.2 percent flap detachment, was 12.2%. A modified Whitehead technique has become the authors' procedure of choice for circumferential prolapsing and bleeding hemorrhoids and mucosa. PMID- 3042302 TI - Evaluation of endorectal ultrasound for the assessment of wall invasion of rectal cancer. Report of a case. AB - To accurately assess the depth of cancer invasion, endorectal ultrasound was performed using a radial scanner (Aloka, 7.5 MHz) in 145 patients with rectal cancer. High-resolution ultrasound clearly depicted five- or seven-layer echographic structures in the normal rectal wall, and demonstrated cancer as a hypoechoic lesion. These layer structures provided an important feature in determining the depth of cancer invasion. Rectal cancers of 122 patients were examined thoroughly by endorectal ultrasound. In 95 of these patients (77.9 percent), ultrasonic assessment of the depth of cancer invasion as classified in four groups was correct, corresponding accurately to the microscopic findings. Ultrasonography overestimated the depth of cancer invasion in 21 patients, however, and underestimated it in six patients. This study indicated that a cause of the overestimation was inflammatory cell infiltration around cancer, and that one possible cause of underestimation was microscopically minimal invasion of cancer. Although there are certain limitations of endorectal ultrasound, this ultrasound technique will provide valuable information to determine the preoperative staging of rectal cancer. PMID- 3042303 TI - Reconstruction of the anus, rectovaginal septum, and distal part of the vagina after postirradiation necrosis. Report of a unique case. AB - Successful repair of postirradiation total loss of the anal sphincters, rectovaginal septum, and distal part of the vagina is reported. Gracilis muscle flap was used as a substitute sphincter. Part of the muscle was "wrapped-up" in a split skin graft. To the authors' knowledge, this is the first report on new application of gracilis muscle and split skin graft in perineal reconstruction. PMID- 3042304 TI - Colorectal cancer. The bases for a comprehensive follow-up. AB - The purpose of this article was to review the effectiveness of follow-up in patients with colorectal cancer submitted to curative treatment. A comprehensive follow-up involves rational initial management of the primary tumor, knowledge of prognostic factors, selection of the patient to be followed, determination of the time for follow-up, use of the most appropriate tests for early diagnosis of recurrence, and eventual curative treatment. The updated answers to all these questions are given through an extensive review of the world literature and confronted with the authors' experience of eight years of follow-up in a series of 170 colorectal cancer patients treated for cure. Although the future might be more promising, past world experience suggests only a few patients could be saved. It is concluded that there is no place for incomplete and disperse screening tests, and only comprehensive, intensive, and very well-coordinated follow-up programs should be undertaken if better results are hoped to be achieved. PMID- 3042305 TI - Lester R. Dragstedt 1893-1975. Chronic ulcerative colitis. A summary of evidence implicating Bacterium necrophorum as an etiologic agent. AB - Lester Dragstedt was born in Anaconda, Montana, the son of Swedish immigrant parents. His entire college and professional education took place at the University of Chicago, where he received a B.S. degree in 1915, a master's degree in physiology in 1916, a Ph.D. in physiology in 1920, and the M.D. degree (from Rush) in 1921. His first academic appointment was as a physiologist at the State University of Iowa. In 1925 Dragstedt was recruited by Dallas B. Phemister to help design the new University Hospital research facilities on the campus of the University of Chicago. Following completion of this responsibility Phemister appointed Dragstedt as Associate Professor of Surgery, stating, "I can teach surgery to a physiologist; I am interested in teaching physiology to surgeons." In 1947 Dragstedt succeeded Phemister as chairman, a post he occupied until his retirement in 1959. Dragstedt was regarded as a skilled clinician as well as a dexterous and artistic surgeon. But he was particularly recognized for his contributions as physiologist-surgeon in the treatment of diseases of the pancreas, parathyroids, and especially diseases of the stomach. In 1943, he performed a transthoracic vagotomy on a patient with a duodenal ulcer who refused to accept the standard operation, subtotal gastrectomy. A lesser known but classical work of Dragstedt and his coworkers is reproduced here for this series. Dragstedt was the originator of the skin-grafted ileostomy in the treatment of ulcerative colitis. The author describes a complete "take" of the split-thickness graft in four patients, although he observed that the "resulting ileostomy looked somewhat like a penis." One can only surmise about the psychologic disability that would be produced. The stoma could, however, be fitted with an appliance that would minimize the risk of abdominal wall digestion. When reading the article and understanding the experimental studies proposing the possible causative organism of ulcerative colitis, one is impressed by Dragstedt's creative thinking. Dragstedt's renown as a basic scientist was illustrated by his election to the National Academy of Sciences. Following his Chicago retirement he became again a full-time physiologist with appointments as research professor in both the department of surgery and the department of physiology at the University of Florida College of Medicine. Active until the end, he died at his summer home on Elk Lake, Michigan on July 16, 1975. PMID- 3042306 TI - Hyperglycemia after intense exercise in IDDM subjects during continuous subcutaneous insulin infusion. AB - Exercise is conventionally considered a modality for improvement of glycemia in diabetes. We have found that a short period of intense exercise (80% VO2max) in normal lean subjects produces sustained postexercise hyperglycemia 20% above basal with a corresponding 100% increase in plasma insulin. In people with insulin-dependent diabetes mellitus (IDDM) incapable of this insulin response, it was predicted that postexercise hyperglycemia would be of greater magnitude and/or duration. To investigate this possibility, the effects of the same intense exercise (80% VO2max) were studied in 8 IDDM subjects (2 on 2 occasions) in the postabsorptive state with continuous subcutaneous (abdominal) insulin infusion (CSII). When the preexercise plasma glucose was normal (n = 6, 86 +/- 4 mg/dl), there ensued a postexercise hyperglycemia to 127 +/- 7 mg/dl (P less than .001) sustained for 2 h postexhaustion. Plasma free immunoreactive insulin (IRI) was 1.43 +/- 0.12 ng/ml before exercise and did not change postexercise. When mean preexercise plasma glucose was 149 +/- 9 mg/dl (n = 4), it rose progressively throughout the 2 h of recovery to 229 +/- 28 mg/dl (P less than .025). A small but statistically significant decrease in free IRI occurred during the last 80 min of recovery. Hyperglycemia in the diabetic subjects was not explained by abnormal or differing responses of glucagon or catecholamines. Thus, with intense exercise, diabetic control deteriorates rather than improves. Therefore, different therapeutic strategies may be required for intense compared with moderate exercise in IDDM patients. PMID- 3042307 TI - Glucagon-stimulated and postprandial plasma C-peptide values as measures of insulin secretory capacity. AB - Basal, postprandial (2 h after breakfast), and glucagon-stimulated plasma C peptide concentrations were determined in a group of 36 adult diabetic patients. Basal and postprandial C-peptide values were measured on consecutive days to estimate the degree of variation of C-peptide secretion. In a subgroup of 15 diabetic patients treated chronically with diet and oral hypoglycemic agents (sulfonylureas or a combination of sulfonylureas and metformin), we studied whether administration of sulfonylureas immediately before breakfast had any effect on postprandial C-peptide values. Absolute differences between two consecutive fasting C-peptide concentrations in insulin-requiring patients were less than 0.1 nM in all but 1 patient, in whom the difference was 0.18 nM. In subjects treated with oral hypoglycemic agents the median difference was 0.12 nM (range 0-0.38 nM). Absolute differences between two consecutive postprandial C peptide concentrations were all less than 0.1 nM in insulin-requiring patients. No significant difference was found between postprandial C-peptide concentrations with or without preceding administration of oral hypoglycemic agents (medians 1.35 and 1.30 nM, respectively). Glucagon-stimulated C-peptide concentrations were somewhat higher than the postprandial values. However, equal discrimination between insulin-requiring and non-insulin-requiring diabetic patients was found by measuring postprandial or glucagon-stimulated C-peptide concentrations. PMID- 3042308 TI - Comparison of plasma glucose and insulin responses to mixed meals of high-, intermediate-, and low-glycemic potential. AB - Although plasma glucose and insulin responses have been shown to vary considerably when either normal subjects or patients with non-insulin-dependent diabetes mellitus (NIDDM) consume different carbohydrate-rich foods, it has been difficult to demonstrate this phenomenon when the same foods have been incorporated into a single mixed meal. To pursue this issue further, plasma glucose and insulin concentrations were determined at hourly intervals from 0800 to 2100 h in NIDDM patients in response to three meals (breakfast, lunch, and dinner) calculated to be of low-, intermediate-, and high-glycemic potency. The total integrated glucose response (mean +/- SE) during the day the low-glycemic meals were ingested was approximately 7% lower (2500 +/- 246 mg.dl-1.h-1) than on the days patients ate either the intermediate- (2701 +/- 280 mg.dl-1.h-1) or high (2718 +/- 311 mg.dl-1.h-1) glycemic meals. When these data were analyzed by meal, it became apparent that the plasma glucose response to breakfast and dinner were essentially identical after consumption of the meals of either low-, intermediate-, or high-glycemic potency. Thus, the modest attenuation of the day long glycemic response on the day patients ate the low-glycemic meal was due to a reduction in plasma glucose concentration after lunch. The day-long plasma insulin responses to the meals of different glycemic potency were qualitatively similar.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3042309 TI - Dietary carbohydrate, a Big Mac, and insulin requirements in type I diabetes. AB - Using the artificial beta-cell (Biostator), we determined the insulin requirements in five nonobese type I (insulin-dependent) diabetic subjects who received isocaloric 40 and 60% mixed-carbohydrate diets in a crossover randomized fashion for 4 days, each day consisting of four equal meals. This was followed on day 5 by a "Big Mac Attack" lunch consisting of a Big Mac, french fries, and milk shake. Insulin requirements to maintain normoglycemia were calculated for each 24 h period and for the 2 h after each meal. The mean 24-h insulin requirements to maintain normoglycemia was greater for the 60% carbohydrate diet than the 40% diet. Although the four meals were of equal size, in all patients the insulin required to cover breakfast greater than lunch greater than dinner greater than or equal to snack. Expressed as milliunits per kilocalorie, the amount of insulin to cover breakfast was greater for the 60% (P less than .05) than the 40% carbohydrate diet and greater for breakfast than the other meals (P less than .01). Insulin requirements for the Big Mac (43% carbohydrate) were 58% greater than for the 40% carbohydrate diet, even after correction for caloric differences. In summary, 1) increasing dietary carbohydrate from 40 to 60% results in an increased insulin requirement for meals only; 2) insulin requirements are greater in the morning than in the evening, even when meal size is constant; and 3) very large meals with high fat and carbohydrate content result in a major increase in insulin requirement. These data indicate that diet has an important impact on insulin requirements in diabetes. PMID- 3042311 TI - Measurement of pulse reappearance time in diagnosis of peripheral vascular disease in diabetes. AB - Forty-eight patients (20 diabetic, 28 nondiabetic) with angiographically confirmed peripheral vascular disease (PVD) were examined to discover whether the measurement of pulse reappearance time (PRT) during reactive hyperemia is a more useful method than the measurement of peripheral systolic blood pressure (ankle pressure index; API) for making a specific diagnosis of PVD. Specific diagnosis refers to the degree and localization of occlusive atherosclerosis determined by Doppler ultrasound techniques for both measurements. We found that PRT and API both provided accurate qualitative proof of a peripheral blood flow deficit in diabetic and nondiabetic subjects. However, in relation to the angiographically defined degree and localization of sclerotic lesions, there were significant differences. The sclerotic degree of occlusive PVD in diabetic subjects was correlated with the results of the PRT (P less than .001), whereas the API was not (P greater than .05). The occlusion localization could only be distinguished by PRT measurements in both diabetic and nondiabetic subjects. Compared with control subjects (4.1 s) the half-maximum PRT of blood flow velocity was delayed in stenotic PVD to 5.7 s, in occlusive PVD of the upper leg to 14.3 s, in occlusive PVD of the lower leg to 29.6 s, and in multilevel disease to 45.0 s (P less than .0005 vs. control). The results show that Doppler sonographic measurement of the peripheral systolic blood pressure is only useful for an overall diagnosis of PVD in diabetic subjects, whereas PRT measurement, by quantifying the degree and localization of sclerotic lesions, can be used additionally either to confirm or to specify this diagnosis. PMID- 3042310 TI - Effects of BAYm 1099, new alpha-glucosidase inhibitor, on acute metabolic responses and metabolic control in NIDDM over 1 mo. AB - To examine the clinical role of BAYm 1099, 15 diet-treated non-insulin-dependent diabetic (NIDDM) subjects were randomized to start drug (50 mg 3 times/day) or placebo after a 4-wk run-in period in a double-blind crossover study. Treatment periods (4 wk) were separated by a 2-wk washout period. During the last week of each treatment period, three test meals (TMs) were administered: 60 g starch (TM1), 25 g sucrose (TM2), and combined 60 g starch and 25 g sucrose (TM3). Twelve subjects completed the study. The peak postprandial blood glucose, lactate, and pyruvate levels (means +/- SE) were significantly lower with active drug after all test meals, particularly TM2 (11.3 +/- 1.0 vs. 14.3 +/- 1.4 mM, P less than .001; 1.53 +/- 0.20 vs. 2.48 +/- 0.17 mM, P less than .001; and 105.1 +/- 17.6 vs. 147.6 +/- 11.1 microM, P less than) less than .001; and 105.1 +/- 17.6 vs. 147.6 +/- 11.1 microM, P less than .05, respectively. Peak blood glucose levels were significantly delayed. However, fasting blood glucose, HbA1, fructosamine, and cholesterol did not change during active treatment (10.0 +/- 1.0 vs. 9.9 +/- 1.0 mM, 10.0 +/- 0.7 vs. 9.4 +/- 0.7%, 2.44 +/- 0.10 vs. 2.37 +/- 0.07 mmol/100 g protein, and 6.7 +/- 0.3 vs. 6.5 +/- 0.3 mM, P NS). Flatulence and diarrhea were severe in 2 subjects, requiring termination of study. Thus, in NIDDM, BAYm 1099 was effective in diminishing and delaying postprandial excursions of blood glucose, lactate, and pyruvate after high- and low-sucrose meals, but overall metabolic control remained unchanged.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3042312 TI - Suspected allergy to insulin preparations. PMID- 3042313 TI - Medical plastics and insulin-pump treatment. PMID- 3042314 TI - Use of NovoPen in diabetic patients on CAPD. PMID- 3042315 TI - Impact of SMBG on control of diabetes as measured by HbA1. 3-yr survey of a juvenile IDDM clinic. AB - Three hundred twelve diabetic children and adolescents were seen in our diabetic clinic and instructed to test their capillary blood glucose (CBG) twice daily and to use an algorithm to adjust their short-acting insulin. Of this group, 219 youngsters had a full 3-yr period of observation. At each clinic visit, blood was obtained for fasting blood glucose and HbA1 and, once a year, cholesterol and triglycerides were also measured. Patient and parent accuracy in measuring CBG was found to be adequate. The changes over time in HbA1 were nondifferential across age and sex, and there was no difference in the level of HbA1 between age and sex groups, the number of tests reported to have been done by the patients, the number of injections of insulin per day, or the serum cholesterol. There was a significant relationship between the HbA1 and the fasting blood glucose (P less than .001) measured by the laboratory as well as with the serum triglyceride (P less than .01). The failure to improve diabetic control, despite measures that would have been expected to do so, was believed to relate more to a lack of compliance than to a flaw in the therapeutic approach. It was interesting to note that the adolescent patients in the study were in no worse control than the younger children in the group. Although better technical skills are available today to manage diabetes, the psychosocial approach to patient motivation requires improvement. PMID- 3042316 TI - Salivary insulin in normal and type I diabetic subjects. AB - We studied the relationship of salivary insulin to serum insulin concentrations in normal subjects and type I (insulin-dependent) diabetic patients to test the hypothesis that salivary insulin might be a simple measure of insulinemia in diabetes. In 8 nondiabetic subjects, salivary insulin levels increased after an oral glucose load but with a delay in peak concentrations of approximately 45 min in comparison with serum insulin levels. There was a significant correlation (r = .810, P less than .01) between mean serum insulin and the salivary insulin 30 min later. In 12 type I diabetic patients, day profiles of saliva and serum insulin were obtained during usual insulin treatment, diet, and physical activity. In serum, the mean (+/- SE) percentage of bound insulin was 58.8 +/- 5.2%, and in saliva it was 45 +/- 3.5%. The mean ratio of salivary to serum free insulin throughout the day was 1:1.6. Although there was a significant correlation (r = .913, P less than .001) between mean serum free insulin for all patients and the corresponding mean free salivary insulin, several individual profiles showed marked discrepancies between the timing and magnitude of insulin changes in the two compartments. We would not, therefore, recommend salivary insulin concentrations as a reliable index of insulinemia in individuals with type I diabetes. PMID- 3042317 TI - Improvement of glucose tolerance in NIDDM by clofibrate. Randomized double-blind study. AB - A randomized double-blind study was performed to examine the effect of clofibrate on glucose tolerance in subjects with non-insulin-dependent diabetes mellitus (NIDDM). Clofibrate (1.5 g/day) or placebo was administered to 70 patients and an oral glucose tolerance test (OGTT) was performed before and 12 wk after treatment. Blood glucose levels were significantly improved in clofibrate-treated groups at all time points during OGTT, whereas there was no change in insulin levels. Improvement of fasting glucose levels required 8 wk of clofibrate treatment. Insulin binding to erythrocytes demonstrated no significant change in the clofibrate-treated subjects. These results suggest that clofibrate improves glucose tolerance in NIDDM subjects without a change in insulin receptors and that clofibrate increases insulin sensitivity through an unknown postreceptor mechanism. PMID- 3042318 TI - DNA polymorphism of the insulin gene, diabetes, and severe obesity. PMID- 3042320 TI - Efficient secretion and processing of heterologous proteins in Saccharomyces cerevisiae is mediated solely by the pre-segment of alpha-factor precursor. AB - A novel processing site was identified in fusions of the alpha-factor precursor of Saccharomyces cerevisiae following its 19 amino-terminal residues (pre segment). Fusions of the pre-segment to heterologous proteins, including aminoglycoside phosphotransferase (APH) and human granulocyte-macrophage colony stimulating factor (hGM-CSF), were as efficiently secreted and processed as corresponding pre-pro fusions. Pre- and pre-pro fusions to hGM-CSF were identically N- and O-glycosylated. While pre-pro fusions to interleukin-1 beta were not cleaved, pre-fusions were correctly processed during secretion. The high secretion efficiency of pre-fusions suggests that the pro-segment of the alpha factor precursor is not required for efficient secretion and processing of protein fusions. PMID- 3042319 TI - Identification and characterization of cryptic polyadenylation sites in the 3' region of a pea ribulose-1,5-bisphosphate carboxylase small subunit gene. AB - The polyadenylation signal of a pea ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit (rbcS) gene has been studied using in vitro mutagenesis and Ti plasmid-mediated transformation of tobacco. Analysis of a mutant that is lacking sequences upstream from -6 (relative to the "normal" site of polyadenylation of RNAs from the rbcS-E9 gene) reveals a number of alternate polyadenylation sites located downstream from the normal poly(A) site. Transcripts carrying these sequences end at any one of at least seven sites between 20 and 300 bases downstream from the normal site. These sites are seen in populations of transgenic plant cells, and also in independent transgenic plants. PMID- 3042322 TI - For Harold E. Henkes--editor-in-chief for Documenta Ophthalmologica 1970-1986. PMID- 3042321 TI - Henkes and the physicist or 40 years of interaction. PMID- 3042323 TI - Retinal dystrophy and macular coloboma. AB - Seven cases of retinal dystrophy associated with bilateral macular colobomata are presented. Two separate entities were found. The first is a congenital onset pigmentary retinopathy similar in electrophysiologic findings and symptoms to typical Leber's congenital amaurosis; the second appears to be a form of pregressive cone-rod dystrophy with pigmentary retinopathy. Review of the pertinant literature and clinical evidence suggest that both conditions are distinct entities inherited in the autosomal recessive manner. PMID- 3042324 TI - Rod densitometry in night blindness: a review and two puzzling cases. Rod densitometry in night blindness. AB - Since the non-invasive technque of retinal densitometry became available in 1955, rhodopsin kinetics could be studied in vivo. It was obvious that with this new tool investigators focussed attention on the aetiology of night blindness in various diseases. A brief review about the clinical developments in the past two decades is given. Also three case-reports are presented, which suggest that in some cases of congenital stationary night blindness (CSNB) the night blindness might arise from the absence of rhodopsin. This is contrary to the standing opinion and present problems regarding the integrity of the retina. PMID- 3042326 TI - Insulin therapy for NIDDM. Edited papers based on a workshop. Geneva, Switzerland, 17 October 1986. PMID- 3042325 TI - Blindness and the eye in mythology and religion as represented on postage stamps. PMID- 3042327 TI - Should non-insulin-dependent diabetics with neuropathy be treated with insulin? PMID- 3042328 TI - Should NIDDM with microvascular disease be treated with insulin? PMID- 3042329 TI - Insulin therapy in the elderly type 2 diabetic patient. PMID- 3042330 TI - When to start an NIDDM patient on insulin. Which insulin programme could be recommended? PMID- 3042331 TI - Diagnosis and clinical aspects of secondary failure of oral antidiabetic agents. PMID- 3042332 TI - Insulin injections: mistakes and errors made by patients and/or health care providers. PMID- 3042333 TI - Secondary failure of oral antidiabetic and dietetic therapy in non-insulin dependent diabetes mellitus. Remission through short sessions of continuous intravenous insulin infusion. AB - The infusion of periodic intravenous insulin for the equilibrium of the diabetic state is proposed for cases of non-insulin-dependent diabetes mellitus (NIDDM) that have become resistant to oral treatment. In order to re-establish the efficacy of oral antidiabetic treatment, 37 patients with NIDDM presenting secondary failure to diet and oral antidiabetic therapy were subjected to sessions of continuous intravenous insulin infusion, resulting in transitory normal blood sugars. With a diminution of symptoms, an increase in the efficacy of oral treatment was noted in 18 cases (48.6%), allowing the continuation of treatment without disturbance of the equilibrium over periods of 6 and 12 months. This improvement is not concurrent with the rise in glucagon-stimulated insulin secretion as evidenced by C-peptiduria and basal C-peptidemia. An improvement in insulin sensitivity (not investigated in this study) might explain this beneficial effect. Periodic intravenous infusions of insulin, based on the diabetic equilibrium, are proposed for the treatment of NIDDM patients that have become resistant to oral therapy. PMID- 3042334 TI - Identification and treatment of patients receiving 'abusive' insulin therapy. PMID- 3042335 TI - Atherogenicity of insulin. PMID- 3042337 TI - Hypoglycemia--mechanisms and prevention in NIDDM treatment with insulin. PMID- 3042336 TI - Insulin resistance in non-insulin-dependent diabetes mellitus (NIDDM). PMID- 3042338 TI - Facts about combination therapy in NIDDM: insulin + oral antidiabetic agents. AB - Recent recommendations in textbooks suggest the use of the combination of insulin treatment with oral antidiabetic agents in NIDDM, especially in secondary failures and insulin resistance with exogenous insulin. This new view in treatment policy is based on literature data in C-peptide positive IDDM patients. Better metabolic control with lower exogenous insulin dosages could also be obtained in NIDDM. If no medical contraindication for drug treatment exists (liver or renal insufficiency, drug interactions, etc.), a combination therapy trial can be considered as an intermediate step between oral antidiabetic agents alone and insulin as monotherapy. Long-term maintenance of endogenous secretion and limited peripheral hyperinsulinism can be considered as potential benefits of this combination therapy. PMID- 3042340 TI - Pathogenesis of NIDDM. PMID- 3042339 TI - The combined treatment with insulin and sulfonylurea in non-insulin-dependent diabetic patients with secondary failure. Rationale and guidelines. PMID- 3042341 TI - Recent status of insulin treatment of Japanese NIDDM. PMID- 3042342 TI - Fetal outcome in gestational diabetes with elevated amniotic fluid insulin levels. Dietary versus insulin treatment. AB - Of 228 women with gestational diabetes between 28 and 32 gestational weeks, 195 had a normal amniotic fluid insulin level (4.8 +/- 3.6 microU/ml) while 33 (14.5%) had an elevated level (23.1 +/- 10 microU/ml). Women with a normal amniotic fluid insulin level were treated by diet alone. Fourteen of the women with an elevated level were treated by diet alone; 19 received insulin treatment additionally. The fetal outcome of patients with a normal amniotic fluid insulin level and dietary therapy and of those with an elevated level and insulin treatment was similar to that of metabolically healthy women. The newborns of gestational diabetics with elevated amniotic fluid insulin treated by diet alone showed a significantly higher incidence of neonatal hyperinsulinism, hypoglycemia, hyperbilirubinemia, high birth weight, respiratory distress syndrome and hypocalcemia. While 2/14 (14%) of the neonates in the dietary group had fatal respiratory distress syndrome, there were no deaths in the group with elevated amniotic fluid insulin and insulin treatment. The data demonstrate that in gestational diabetics with normal amniotic fluid insulin (low-risk group), dietary therapy is sufficient while insulin therapy is required to ensure healthy offspring in patients with elevated amniotic insulin (high-risk group). PMID- 3042343 TI - A comparison of cellular actions between gliclazide and a hypoglycaemic peptide fragment of human growth hormone (hGH 6-13). AB - The mechanisms of hypoglycaemic action of a 'second-generation' sulphonylurea, gliclazide, and a synthetic human growth hormone fragment, hGH 6-13 (Leu-Ser-Arg Leu-Phe-Asp-Asn-Ala), were compared at the cellular level in rats. Both compounds were shown to be hypoglycaemic in vivo although their molecular structures were totally different. Gliclazide was markedly insulinotropic, as are all hypoglycaemic sulphonylureas, whereas hGH 6-13 had no visible effect on basal levels of plasma insulin. However, in vitro studies with isolated pancreatic islets revealed that hGH 6-13 significantly augmented insulin secretion in the presence of exogenous glucose. One other major difference was that gliclazide had no direct effect on insulin receptor function while the synthetic hGH 6-13 increased the binding of insulin to specific receptors on isolated cells. Results suggested that the human growth hormone fragment hGH 6-13 could be a potential anti-diabetes drug with the ability to potentiate circulating insulin action and to achieve blood glucose normalisation. PMID- 3042345 TI - Effect of chloroquine on biosynthesis, release and degradation of insulin in isolated islets of rat pancreas. AB - Insulin has been reported to degrade inside the islets and islet lysosomal proteases have been thought to take part. As chloroquine is regarded as a potent lysosomotropic agent, an attempt has been made to see whether chloroquine has an influence on intrainsular degradation of insulin. After preculture of collagenase isolated rat islets at 11 mM glucose together with [3H]leucine for 3 days for labelling newly synthesized insulin, islets were cultured for 1 day at 2.2 or 22 mM glucose with or without 0.02 mM chloroquine. Afterwards, radioactivity was measured in the proinsulin/insulin fraction. For control, the influence of chloroquine during 3-h incubation of both freshly isolated and precultured islets was also studied. During the 1-day culture at 2.2 mM glucose, prelabelled insulin was degraded significantly and addition of chloroquine did not alter the amount of insulin degraded. At 22 mM glucose, no significant amount of insulin had been degraded. During the 3-h incubation of freshly isolated as well as precultured islets, chloroquine was found to inhibit significantly glucose-induced biosynthesis of insulin. Glucose-induced release of insulin, however, was not influenced by chloroquine. It is concluded that chloroquine does not influence glucose-induced release or intra-insular degradation of insulin, but it interferes with the biosynthesis of insulin. PMID- 3042346 TI - Laxatives: replacing danthron. PMID- 3042344 TI - Non-insulin-dependent diabetic patients (NIDDMs) do not demonstrate the dawn phenomenon at presentation. AB - A dawn rise of plasma glucose (PG) and/or insulin, the 'dawn phenomenon', has been commonly reported in treated diabetic patients and normal subjects. To evaluate the effect of treatment on this phenomenon in non-insulin-dependent diabetics (NIDDMs), PG, C peptide, immunoreactive insulin (IRI), growth hormone (GH), cortisol, epinephrine, and norepinephrine were measured hourly between 24.00 and 09.00 h in 17 newly diagnosed untreated NIDDMs (group 1). The study was repeated in 11 patients after a year of treatment (group 2). The PG levels did not change significantly at any time from 03.00 to 08.00 h in group 1 but increased continuously from 6.7 +/- 0.5 mmol/l at 04.00 h to 7.8 +/- 0.5 mmol/l at 08.00 h (P less than 0.01) in group 2. IRI and C peptide decreased significantly after 07.00 h in both groups. GH and catecholamine changes were similar in group 1 and group 2. Cortisol levels showed a nadir at 02.00 h and a peak after 07.00 h in both groups. Our results demonstrate no dawn rise of mean PG, IRI and C peptide in newly diagnosed untreated NIDDMs but a significant rise of PG in the early morning period in NIDDMs after a year of treatment with diet alone and diet plus sulphonylureas. Therefore other factors such as treatment and/or duration of the diabetes may play an important role in the pathogenesis of the dawn phenomenon. PMID- 3042347 TI - [EDRF--the endogenous nitrate vasodilator]. PMID- 3042348 TI - [Therapeutic plasma exchange in the hemolytic-uremic syndrome]. PMID- 3042349 TI - [Insulin: a cardiovascular risk factor?]. PMID- 3042350 TI - Procainamide induces a transitory impairment of B cell mitogenesis in beagle dogs. AB - Beagle dogs (3 to 6 years old) were treated with 100-150 mg procainamide HC1/kg/day. After 2, 5, and 9 months of treatment, peripheral blood lymphocytes were isolated and stimulated with pokeweed mitogen. The data demonstrated a suppression of mitogenesis only at 2 and 5 months after procainamide treatment. The lymphocytes from dogs treated for 9 months had a normal response to pokeweed mitogen. At no time during this experiment were any significant levels of serum antinuclear antibodies detected nor was any change in the number of cycling lymphocytes apparent in the experimental versus control groups. The resting membrane potential of both control and experimental groups was similar and pokeweed mitogen depolarized the cells from both groups. PMID- 3042352 TI - Hypertension and diabetes. Clinical problems. AB - The choice of an appropriate antihypertensive agent and the hazards of postural hypotension are common problems faced in the treatment of diabetic hypertensive patients. The results of 3 studies addressing these problems are described in this report. In the first study, indoramin, an alpha-blocking agent, was administered to patients with non-insulin-dependent diabetes and mild to moderate hypertension. Blood pressure control was achieved in 57% of patients with mild, and in none with moderate hypertension. The blood glucose and insulin responses to an oral 50g glucose loading, as well as the blood concentrations of HbA1 did not change during therapy. Seven patients were excluded because of side effects. In 4 of them postural hypotension was observed. In the second study, the effects of angiotensin-converting enzyme (ACE) inhibitors, administered to patients with non-insulin-dependent diabetes and mild to moderate hypertension, were evaluated. Blood pressure control was achieved in 78% of the patients on captopril (n = 14) and in 74% of patients on enalapril therapy (n = 23). Symptomatic postural hypotension (n = 2) and hyperkalaemia (n = 2) were observed with both drugs. Significant reductions in 24-hour urinary protein or albumin excretion were detected in 12 patients on enalapril therapy. No changes in 2-hour postprandial blood glucose and HbA1 levels were observed during therapy with ACE inhibitors. In the third study, dopaminergic antagonist agents were evaluated in diabetic patients with orthostatic hypotension. In 7 patients metoclopramide (20mg intravenously) reduced the fall in mean arterial pressure induced by upright tilt.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3042353 TI - Responders and non-responders to antihypertensive treatment. AB - Responders to antihypertensive treatment have been defined as any patient with a fall in mean arterial pressure, however small; any patient with a fall in mean arterial pressure of 5%, 10% or greater; and any patient achieving a goal blood pressure, usually a diastolic pressure less than 90 mm Hg. Non-responders are normally defined as those who do not fit into the category of a responder. Blood pressure, however, varies considerably during follow-up, and diastolic pressure has a within-subject standard deviation of 8 mm Hg. Blood pressure varies according to environmental factors such as temperature, diet and stress, and tends to drift downwards during follow-up as the patient becomes accustomed to having blood pressure taken, to the place of measurement and to the observer. In a statistical sense, responders may be defined as the 2.5% with a fall in DBP greater than 16 mm Hg, yet in clinical studies of a low sodium diet reports have indicated that 50% have experienced an increase in pressure and 50% a fall (responders). Although the characteristics of those with a fall in pressure are of interest and should be compared with those who do not respond, a 50:50 division is unlikely to help. Methods of identifying true responders and non responders are discussed in this brief review, along with the errors that may arise from a misclassification and problems of conducting further trials of treatment on the non-responders. PMID- 3042355 TI - Double-blind comparison of urapidil and prazosin in the treatment of patients with essential hypertension. AB - A multicentre double-blind comparative study of urapidil and prazosin was performed in 412 outpatients with mild to moderate essential hypertension. 222 patients were treated by monotherapy (107 on urapidil and 115 on prazosin), and 190 in combination with thiazide diuretics (99 on urapidil and 91 on prazosin). After a 4-week control period on placebo, the patients were randomised for treatment with twice daily urapidil (total of 30-120 mg/day) or 3 times daily prazosin (total of 1.5 to 6 mg/day) for 12 weeks. The results indicated that: (1) In monotherapy, blood pressure decreased significantly in both treatment groups 2 weeks after the start of treatment, and a further decrease in blood pressure was observed in both groups thereafter. The rates of responders (final mean blood pressure reductions of 13 mm Hg or more) were similar: 64.1% in the urapidil group and 64.0% in the prazosin group. Heart rate did not change in the 2 groups at any stage. Side effects were observed in 15.9% of the patients in the urapidil group and in 11.3% of the prazosin group (NS). When an overall utility rate was determined on the basis of antihypertensive effect, side effects and laboratory findings, the treatment was judged 'useful or better' in 58.9% of the patients in the urapidil group and 60.2% of the prazosin group. The difference was again not significant. (2) In combination therapy, a significant decrease in blood pressure was also observed 2 weeks after the start of treatment. The responder rates were 66.7% in the urapidil group and 65.0% in the prazosin group (NS).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3042354 TI - Efficacy of urapidil in the management of essential hypertension. A comparison with captopril. AB - In a multicentre study by general practitioners, the antihypertensive efficacy of urapidil, a postsynaptic alpha-blocker with central action, was compared with that of the angiotensin-converting enzyme inhibitor captopril. The study was of a double-blind, randomised parallel-group design. Following a 2-week washout and placebo phase, 295 essential hypertensives (WHO I/II) were treated for 12 weeks with either urapidil or captopril, initially with urapidil 60 mg twice daily or captopril 25mg twice daily, with the possibility of adjusting the dose according to blood pressure response after 2 weeks of treatment [diastolic blood pressure (DBP) 90 mm Hg or less: reduction of urapidil to 30 mg twice daily or captopril to 12.5mg twice daily; DBP 91 to 99 mm Hg: dose unchanged; DBP 100 mm Hg or more: dose increased to urapidil 90 mg twice daily or captopril 50 mg twice daily]. Blood pressure values at the end of the 12-week treatment period dropped significantly in the urapidil group (n = 142; all dosages) from 175/103 mm Hg to 154/89 mm Hg (p less than 0.001) and in the captopril group (n = 153; all dosages) blood pressure decreased from 175/103 mm Hg to 154/90 mm Hg (p less than 0.001), corresponding to 62% and 58% urapidil and captopril responder rates (DBP less than or equal to 90 mm Hg), respectively. The responder rate at 12 weeks under urapidil therapy was 30% for the initial, 17% for the lower and 15% for the higher dose; the respective values for captopril were 28, 16 and 14%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3042351 TI - Serum lipoproteins during treatment with antihypertensive drugs. AB - Hypertension and certain alterations in serum lipoproteins such as a decrease in high density lipoprotein-cholesterol (HDL-C), an increase in low density lipoprotein-cholesterol (LDL-C) and perhaps also elevated triglycerides (Tg), are complementary coronary risk factors. Moreover, it has become evident that several of the drugs used for standard antihypertensive therapy may also interact with lipoprotein metabolism. The following has been observed after 1 to 12 months of treatment. Various diuretics can significantly increase LDL-C and/or very LDL-C and total C/HDL-C ratio, while HDL-C is often largely unchanged; Tg also are often elevated. LDL-C increased in diuretic-treated men and in chlorthalidone treated postmenopausal women but not in chlorthalidone-treated premenopausal women. The latter may be protected from this side effect. Drug dosages were usually high in these studies. Indapamide, given at a dose of 2.5 mg/day, seems to exert no relevant effect on the lipoproteins. It is not established whether this difference is related to the nature of the drugs or the doses used. There is little doubt that the dose of chlorthalidone used was greater than that required for a full antihypertensive effect of this drug. Several beta-blockers given as monotherapy induce significant increases in Tg and a tendency for decreases in HDL-C. These changes are most prominent on non-selective beta 1+2-blockers without partial intrinsic sympathomimetic activity (ISA), less pronounced on highly selective beta 1-blockers without ISA, and even more discrete or absent on beta-blockers with distinct ISA. Other sympatholytics such as reserpine, methyldopa, debrisoquine, urapidil, clonidine, labetalol, or postsynaptic alpha blockers (prazosin, trimazosin, doxazosin etc.) did not affect or, postsynaptic alpha-blockers in particular, sometimes even slightly decreased Tg or LDL-C and very LDL-C values. During combination therapy, diuretic-induced increases in LDL C were at short term prevented or reversed by the concomitant administration of certain beta-blockers, but not by sympatholytics such as reserpine, methyldopa or clonidine. With combined diuretic-prazosin treatment, a tendency for slightly higher HDL-C was reported. Angiotensin converting enzyme inhibitors (captopril, enalapril) and calcium channel blockers (verapamil, nifedipine, nitrendipine, diltiazem) seem to be largely devoid of undesirable effects on serum lipoproteins. Monotherapy with the potent direct vasodilator carprazidil improved blood pressure and significantly increased HDL-C. Whether and to what extent the observed variations in lipoproteins may persist beyond 1 year of treatment is as yet unclear.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3042356 TI - Comparison of urapidil and atenolol in hypertension. AB - The antihypertensive efficacy of urapidil was compared with that of atenolol in a double-blind randomised parallel-group 10-week study. 44 patients with essential hypertension entered the study, and 43 with a supine DBP of 100 to 125 mm Hg after a 2-week placebo period were randomised either to urapidil (n = 22; 60 to 120 mg/day) or atenolol (n = 21; 50 to 100 mg/day) for 8 weeks. Blood pressure, heart rate, electrocardiogram (supine and standing) and side effects were recorded every week, and medical examinations and laboratory tests were done at the end of the placebo and active treatments. 36 patients completed the trial (17 on urapidil and 19 on atenolol). Patients' characteristics were similar in the two groups, and there were no differences between them in blood pressure and heart rate at the end of placebo period. Both drugs produced a significant fall in supine and standing blood pressure; urapidil decreased supine blood pressure from 164/109 to 150/96 (p less than 0.001), and atenolol decreased it from 167/111 to 146/94 mm Hg by week 8 (p less than 0.001). There were no differences in the blood pressure-reducing effects of the two drugs. There was no change in heart rate in the urapidil group, but a marked reduction occurred in the atenolol group, with significant differences from placebo and urapidil (p less than 0.001). Side effects were reported in 32% of the urapidil group and in 29% of the atenolol group; however, these were mild and transient in all but two patients (urapidil = 1, atenolol = 1) who were withdrawn.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3042357 TI - Guidelines for the treatment of mild hypertension. AB - Mild hypertension is defined as a state in which diastolic blood pressures ranging from 90 to 104 mm Hg are persistently observed. Systolic blood pressure is increasingly being recognised as a risk factor in its own right. Strictly speaking, the lessons learnt from the prospective trials in mild hypertension apply mainly to the stratum of mild diastolic hypertension. In this review, randomised placebo-controlled blind trials in uncomplicated mild hypertension (USPHS, ANBPS, MRC trials) will serve as the main data base from which to derive practical guidelines, in terms of benefits and risks, of early treatment. PMID- 3042358 TI - Urapidil in the treatment of hypertension. AB - Urapidil is an alpha 1-adrenoceptor antagonist which also has a central antihypertensive effect, the mechanism of which has yet to be conclusively defined. A number of open and comparative studies have produced evidence for the efficacy and safety of urapidil. A study recently completed by the author produced a dose-dependent antihypertensive effect of urapidil which, however, failed to achieve statistical significance, probably due to a large variance of the data and an unexpectedly large placebo effect. Adverse reactions are those expected from an alpha 1-blocker, particularly dizziness, as well as nausea and fatigue. Urapidil is potentially an important new antihypertensive agent; further variable dose and combination (with other antihypertensive agents) studies would help further define its therapeutic niche. PMID- 3042359 TI - Human pharmacology of urapidil. AB - Urapidil is a phenylpiperazine-substituted uracil derivative used in hypertension. It is rapidly absorbed when given by mouth. Peak blood concentrations of the slow release capsule occur 4 to 6 hours after administration. Oral bioavailability is 78% (range 72 to 84%) and distribution half-life and terminal half-life are about 35 minutes and 3 hours, respectively. Plasma clearance is 12 L/h and renal clearance 1.8 L/h. Seventeen percent appears in urine as the parent compound within 24 hours of dosing. There is extensive hepatic metabolism to the parahydroxylated (34% in urine). N-demethylated (4% in urine) and O-demethylated (3% in urine) products. Elimination is not saturable at usual clinical doses. The major action of urapidil is post-synaptic alpha 1 adrenergic blockade, with a minor degree of beta 1-adrenergic blockade and a centrally mediated reduction in sympathetic outflow which has an as yet unidentified basis. PMID- 3042361 TI - Haemodynamic effects of urapidil in arterial hypertension and congestive heart failure. AB - An elevated total peripheral resistance is the haemodynamic hallmark of both arterial hypertension and congestive heart failure. In essential hypertension it is the main pathogenetic abnormality, whereas in congestive heart failure it is the result of vasoconstriction serving to compensate for the fall in cardiac output and arterial pressure. Any drug that lowers total peripheral resistance can, therefore, potentially favourably influence established hypertension as well as unload the left ventricle in congestive failure. In essential hypertension, urapidil lowers arterial pressure acutely by a fall in total peripheral resistance with a small compensatory increase in cardiac output. Concomitantly, renal and splanchnic blood flow increase and the resistance in these vascular beds falls. In congestive heart failure, the acute administration of urapidil increases cardiac index and lowers mean pulmonary artery wedge pressure and systemic vascular resistance by about 30%. At the same time, a mild fall in mean arterial pressure is observed. Long term non-invasive studies document that these acute haemodynamic effects remain preserved in both arterial hypertension and congestive heart failure. Urapidil seems, therefore, to be a promising agent for the treatment of haemodynamic disorders that are characterised by an elevation of total peripheral resistance, such as established essential hypertension and congestive heart failure. PMID- 3042362 TI - The development and progression of chronic renal disease. Can it be prevented or attenuated? AB - Many forces contribute to the immutable progressive deterioration of renal diseases. This review focuses on the pernicious haemodynamic response of the kidney to an initial loss of mass. Afferent arteriolar dilatation, coupled with relative or absolute efferent arteriolar constriction, causes the hydrostatic pressure in the intervening glomerular capillaries to increase. While sustaining the glomerular filtration rate, the glomerular hypertension may ultimately scar and destroy the kidney. Experimental studies in animals persuasively argue that the high glomerular capillary pressures do indeed contribute to progressive renal damage. Whether this observation is translatable into human renal diseases is the subject of ongoing clinical investigation. The role of dietary protein restriction and converting enzyme inhibitors in reducing this glomerular hypertension and in potentially attenuating the progression of a wide range of renal diseases is also discussed. PMID- 3042363 TI - Mechanisms of complete baroreceptor resetting in hypertension. AB - It is a universally accepted phenomenon that vascular baroreceptors reset to operate at higher pressure levels in hypertension. A rapid or acute resetting can be demonstrated within the first 5 to 15 minutes after arterial pressure has been elevated. However, this resetting is only partial because the increase in pressure threshold for baroreceptor activation represents a fraction (20 to 40%) of the total pressure increase. This acute resetting changes little within the first hours of hypertension. To achieve complete resetting however, that is when the pressure threshold increase equals the total pressure increase, blood pressure needs to be maintained at an elevated level for 48 hours in the rat. Studies of aortic calibre behaviour performed in freely moving rats demonstrated a striking direct relationship between the time taken for the diastolic calibre to reach maximal dilation and the time taken for complete resetting of the aortic baroreceptors. It was also observed that during transient pressure increases the displacement of the diastolic calibre is much greater than the increase in pulsation, indicating that under physiological conditions sustained distension of the diastolic calibre is an important factor in aortic baroreceptor distortion. The data suggest that complete resetting of the baroreceptors in hypertension occurs when the increased stress on the arterial wall is matched by a proportional permanent increase in diastolic calibre. PMID- 3042364 TI - An analysis of staging systems for carcinoma of the maxillary sinus. PMID- 3042366 TI - Assessment of a simple method for enhancing bond strength between porcelain and a nickel-base alloy. PMID- 3042365 TI - Shigella boydii and Shigella sonnei: serotype and drug susceptibility patterns in Addis Ababa, Ethiopia (1974-85). PMID- 3042367 TI - Vertical displacement of the mucosa in bounded edentulous ridges with different elastomeric impression materials and techniques. PMID- 3042368 TI - Rationalisation of bridge design. PMID- 3042360 TI - Drug treatment of hypertension. AB - There are several first choices for the treatment of mild and moderate hypertension. The selection of a drug may be influenced by concomitant pathology, with positive indications for particular drugs, e.g. coexistent angina, indicating use of a beta-receptor blocking drug or calcium antagonist; fluid retention indicating a diuretic; or contraindication e.g. asthma, and beta adrenoceptor blocking drugs. beta-Adrenoceptor blocking drugs have the advantage of a long history and of possibly being cardioprotective following myocardial infarction, but they have not yet been established as primary preventive agents in hypertensive patients. The alpha-receptor blocking drugs have the advantage of favourably affecting lipid profile and blood pressure. Therefore, there may be advantages in the use of combined alpha- and beta-blockade. The diuretics, which have the advantage of being inexpensive, are widely used but long term metabolic effects, particularly hypokalaemia, cause concern. This is correctable by co administration of a potassium sparing diuretic and often preventable by using low doses of the diuretic. Diet may be important as hypokalaemia appears to be less of a problem where potassium intake is high. Experience with calcium antagonists is widening but the use of converting enzyme inhibitors is more limited, and some physicians are less ready to use them as first choice in mild hypertension at present. Drugs like methyldopa, clonidine, the adrenergic neurone inhibitory drugs are now used more as reserve agents. More severe cases of hypertension may require drugs from 2 of the 3 major groups: beta-blocking drugs, vasodilators and diuretics. In some cases, drugs from each of these 3 groups will be required. PMID- 3042369 TI - Differentiation of fetal rat somatotropes in vitro: effects of cortisol, 3,5,3' triiodothyronine, and glucagon, a light microscopic and radioimmunological study. AB - Cortisol stimulates somatotrope differentiation in vitro. T3 and/or glucagon may also be involved. Fetal rat pituitary primordia were explanted at 14 days gestation and cultured for 7 days in medium supplemented with cortisol (50-500 nM), and either T3 (0.67 nM) or glucagon (0.5 nM). Also, to determine the time of first appearance of the somatotropes, explants were cultured 4, 5, or 6 days with cortisol alone. Immunoreactive somatotropes were detected by immunohistochemistry, and their size and number were estimated for each medium. GH was measured by RIA in explants and media. Immunoreactive somatotropes first appear at 18-19 days gestation. Their size and number depend on cortisol concentration: no cells at 50 nM, a few small ones at 100 nM, and many large ones at 250-500 nM. This progression was reflected by RIA of GH in explants and media, although small quantities were detected with 50 nM. The effect of T3 was only visible with a low dose of cortisol. With 100 nM cortisol, it increased the size and number of cells. Differentiation was also triggered with 50 nM cortisol plus T3. RIA detected significantly higher GH content and secretion after T3 stimulation. The decreases in number, size, and GH secretion and content elicited by glucagon were not significant, probably due to the high variability. Both techniques used provide similar information on somatotrope differentiation: stimulation by cortisol alone, or alternatively by a synergistic action between cortisol and T3. PMID- 3042370 TI - Pancreastatin: molecular and immunocytochemical characterization of a novel peptide in porcine and human tissues. AB - Pancreastatin, a novel 49-amino acid peptide isolated from porcine pancreas, shows over 70% sequence homology to the central part of bovine and human chromogranin-A. Using an N-terminal and C-terminal synthetic peptide, we developed two sensitive and specific RIAs for the detection of pancreastatin-like immunoreactivity (PLI) in porcine and human tissue extracts. PLI was present throughout the gastrointestinal tract and in most endocrine and neuronal tissues. Highest concentrations were measured in the pituitary, adrenal gland, and pancreas (1200-4000 pmol/g), similar to the distribution of chromogranin-A. PLI was also detected in human endocrine tumors, with large quantities in some carcinoids (up to 14 nmol/g). HPLC revealed that extracts from porcine pituitary and pancreas contained small pancreastatin-like peptides, whereas in adrenal medulla large chromogranin-A-like molecular forms predominated. Human endocrine tumors showed a different pattern, with intermediate forms distinct from chromogranin-A and pancreastatin. Biochemical analysis was confirmed by immunocytochemistry localizing PLI in pancreatic islets, adrenal medulla, pituitary, duodenum, and human endocrine tumors. Pancreastatin is present in a variety of gastrointestinal, endocrine, and neuronal tissues and may represent a novel peptide of unknown physiological function, derived from chromogranin-A by proteolytic cleavage. PMID- 3042371 TI - Insulin and a putative insulin metabolic mediator fraction from liver and muscle stimulate p33 messenger ribonucleic acid accumulation by apparently different mechanisms. AB - We have previously shown that in rat H4 hepatoma cells insulin enhances the nuclear transcription of p33 mRNA in a dose- and time-dependent manner, with no alteration in mRNA half-time (t1/2). Presumably, this effect is mediated by the cell surface receptor. In this report, we have investigated the effect of putative insulin mediator fractions which act to control metabolic events on p33 mRNA accumulation in these cells. Initial experiments originally demonstrated an insulin-like effect of an added putative metabolic fraction to enhance p33 mRNA concentrations. However, when the fetal calf serum supply was changed, the effect of insulin remained, but that of added mediator was no longer observed. After a series of experimental approaches designed to alter the permeability of the cell membrane, it was found that in the presence of increased Ca2+, the effect of mediator could again be observed. The present data demonstrate that the partially purified cAMP-dependent protein kinase/adenylate cyclase inhibitory putative mediator fractions from liver and muscle enhance p33 mRNA accumulation in intact H4 hepatoma cells by a mechanism that is differentiated from that of insulin. The action of the putative mediator is inhibited by cycloheximide, while the action of insulin itself is not. These results suggest that insulin may control nuclear transcription by multiple signaling mechanisms. Alternatively, the added putative metabolic mediator may not enter the cell in the presence of cycloheximide or is inactive as such within the cell and must first be converted to an active species by a step requiring protein synthesis. PMID- 3042372 TI - Effects of the amino-terminal portion of human growth hormone on glucose clearance and metabolism in normal, diabetic, hypophysectomized, and diabetic hypophysectomized rats. AB - A naturally occurring pituitary peptide, human (h) GH-(1-43) potentiates insulin action. The present study has compared the effects of acute (30-60 min) and chronic (3-6 days) injections of synthetic hGH-(1-43), hGH, and insulin in normal, diabetic, hypophysectomized, and diabetic-hypophysectomized rats. Male rats (150-250 g) received injections of saline, insulin (50-200 mU), hGH (200 micrograms), or hGH-(1-43) (200-400 micrograms) with or without insulin. Hormone and glucose were injected simultaneously for glucose tolerance tests. Basal and insulin-stimulated [U-14C]glucose oxidation to 14CO2 in adipose tissue were measured in vitro after in vivo treatments; insulin release by isolated pancreatic islets was determined in vitro. Acute injections of hGH-(1-43) with insulin dramatically increased glucose clearance in diabetic (P less than 0.05) and hypophysectomized (P less than 0.01) rats. In diabetic-hypophysectomized rats acute injections of hGH-(1-43) significantly lowered the elevated basal blood glucose level (P less than 0.025) and stimulated [U-14C]glucose oxidation to 14CO2 in adipose tissue (P less than 0.05); it did not increase the glucose clearance rate during glucose administration. Chronic treatment of diabetic rats with hGH-(1-43) did not lower the elevated blood glucose level significantly, but it stimulated [U-14C]glucose oxidation to 14CO2 in adipose tissue; the oxidation was further stimulated by treatment with insulin. Chronic injections of hGH-(1 43) slightly lowered blood glucose levels in hypophysectomized rats (P less than 0.025) despite a diminished release in vitro of insulin from pancreatic islets (P less than 0.05). Therefore, these experiments show hGH-(1-43) to be an insulin potentiator that increases insulin-stimulated glucose clearance and glucose oxidation without an increase in insulin secretion, and they suggest that the peptide may have a physiological role in regulating carbohydrate metabolism. PMID- 3042373 TI - Nutrient and hormonal regulation of the threshold of glucose-stimulated insulin secretion in isolated rat pancreases. AB - This study demonstrates that the threshold of glucose-stimulated insulin secretion can be regulated in vivo by long term hormonal and nutrient modifications. The sensitivity of the pancreatic B-cell to glucose stimulation was determined by examining the pattern of insulin release from pancreases perfused with linear glucose gradients. Male rats infused with ovine PRL for 4 days and rats receiving five hourly injections of glucose had a lower threshold and enhanced rates of insulin release at all stimulatory glucose concentrations. Infusion of bGH for 4 days was without effect on glucose gradient-stimulated insulin release. Fasting the rats for 48 h resulted in an elevation of the threshold and a substantial reduction in the extent of insulin release. To determine possible processes involved in these long term modifications of the threshold of glucose-stimulated insulin secretion, the in vitro effect of potentiators of insulin release was examined. Forskolin, glucagon, cholecystokinin, and carbamylcholine were able to lower the threshold and increase the extent of insulin release. This suggests that the long term regulation of insulin secretion may modulate processes controlling cAMP concentrations and the hydrolysis of phosphoinositides in pancreatic B-cells. Also, the proposed incretin gastric inhibitory polypeptide was capable of lowering the threshold and increasing insulin secretion at stimulatory glucose concentrations. The consequences of a decreased threshold is a markedly enhanced insulin secretion at normal serum glucose concentrations. PMID- 3042374 TI - Changes in cellular levels of messenger ribonucleic acid encoding gonadotropin releasing hormone in the anterior hypothalamus of female rats during the estrous cycle. AB - Cellular levels of messenger RNA encoding GnRH were measured using quantitative in situ hybridization in coronal sections through the area of the organum vasculosum of the lamina terminalis of female rats examined at various times of the 4-day estrous cycle. GnRH mRNA levels were high on the morning of diestrus day 1, but declined throughout the day of diestrus day 2 to a nadir on the morning of proestrus. Although GnRH message levels were lowest on the morning of proestrus, they rose nearly two-fold by 1900h that evening and remained high during the day of estrus. These data support the hypothesis that GnRH synthesis is coupled to GnRH release, and indicate that GnRH biosynthesis is not stimulated on the morning of proestrus in preparation for the ovulatory surge release of GnRH and LH in the afternoon. PMID- 3042375 TI - Androgen abuse by athletes. PMID- 3042376 TI - Reproductive endocrinology of the mink (Mustela vison). PMID- 3042377 TI - The lung at work. PMID- 3042378 TI - Dietary requirements and athletic performance of horses. AB - There is no clear evidence that the chronic requirement for any non-energy yielding nutrient rises in proportion as the energy requirement increases with hard work. The need for protein, and probably that for calcium, remain a function of bodyweight daily. Some proportionality with energy may exist for certain nutrients, although the evidence has not been adduced. For example, because of an increase in both the proportion and amount of propionic acid in the volatile fatty acids of caecal contents, the tissue requirement for vitamin B12 may rise with an increase in the rate of energy metabolism. Exercise influences appetite and therefore voluntary intake, and consequently the daily intake of nutrients. Although that intake is not just a function of dietary bulk and weight, it is necessary to increase energy concentration of diets to achieve an adequate chronic intake of energy where work intensity and energy expenditure are considerable. Acute nutrient requirements paint a different picture from chronic requirements. An increase in total feed intake, or the density of that feed, would neither satisfy these requirements nor be a desirable means of doing so. The acute needs of water, electrolytes and soluble carbohydrates should be met by dosing when the need arises. The timing of the consumption of energy yielding substrates relative to that of exercise may be critical to performance. An inevitable postprandial consequence of a meal of starch or protein by the resting horse, is an increase in the activity of plasma insulin. This increase decreases blood glucose, depriving muscles of a critical substrate, but the assertion has not been resolved by experiment in horses. Experiments are required to ascertain the optimum feeding regime during the 24 h preceding extreme exertion. Whereas exhaustion in sprint work is largely a function of elevated blood lactate concentration, that of extended work is a consequence of a decline in glycogen reserves and losses of body fluid and electrolytes. Glycogen loading is of benefit to many long distance human athletes, but no advantage has yet been established for this practice in horses, and without modification it could render them subject to laminitis and endotoxaemia. Nevertheless supplementation of horses with water, glucose and electrolytes during work may benefit their endurance. The provision of 5 litres water every 2 h with 30 g salt, or twice as much of mixed electrolytes, and 15 g sucrose or glucose, is recommended for a 500 kg horse during periods of extreme sweating.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3042379 TI - Angular limb deformities in foals. PMID- 3042380 TI - Intervention programmes for work-related neck and upper limb disorders: strategies and evaluation. PMID- 3042381 TI - Site-specific recombination in bacteriophage Mu: characterization of binding sites for the DNA invertase Gin. AB - Site-specific DNA inversion in phage Mu is catalysed by the phage-encoded DNA invertase Gin and a host factor FIS. We demonstrate that purified Gin protein binds specifically to 34-bp sequences that flank the G segment as inverted repeats. Each inverted repeat (IR) contains two binding sites for Gin which have to be arranged in a specific configuration to constitute a recombinogenic site. While one of these sites is bound when present alone, the other site is bound only in conjunction with the first one, suggesting cooperative binding. In addition to the sites within the IR, Gin binds with lower affinity to AT-rich sequences adjacent to the IR. We demonstrate that these sites do not participate in the inversion reaction. The IR itself can be shortened to 25 bp without effect on inversion frequency. Using gel mobility shift experiments on circular permuted fragments containing the IR we show that Gin bends DNA upon binding. We discuss the possibility that DNA bending is related to the formation of a productive synaptic complex. PMID- 3042382 TI - The DNA invertase Gin of phage Mu: formation of a covalent complex with DNA via a phosphoserine at amino acid position 9. AB - The DNA invertase Gin encoded by bacteriophage Mu catalyses efficient site specific recombination between inverted repeat sequences (IR) in vivo and in vitro in the presence of the host factor FIS and the recombinational enhancer. We demonstrate that Gin alone is able to introduce single strand breaks into duplex DNA fragments which contain the IR sequence. Strand cleavage is site-specific and can occur on either strand within the IR. Cleaved molecules contain Gin covalently attached to DNA. The covalent complex is formed through linkage of Gin to the 5' DNA phosphate at the site of the break via a phosphoserine. Extensive site-directed mutational analysis showed that all mutants altered at serine position 9 were completely recombination deficient in vivo and in vitro. The mutant proteins bind to DNA but lack topoisomerase activity and are unable to introduce nicks. This holds true even for a conservative amino acid substitution at position 9. We conclude that serine at position 9 is part of the catalytic domain of Gin. The intriguing finding that the DNA invertase Gin has the same catalytic center as the DNA resolvases that promote deletions without recombinational enhancer and host factor FIS is discussed. PMID- 3042384 TI - Tissue-specific expression of a vimentin--desmin hybrid gene in transgenic mice. AB - We have introduced a hybrid gene, pVVim2, composed of the 5' region of the hamster vimentin gene encoding the head and rod domain of vimentin and the 3' region of the hamster desmin gene encoding the tail domain of desmin, into the germ line of mice by pronuclear injection. RNA and protein analysis of mice transgenic for this construct showed that the pVVim2 gene was expressed at high levels in a developmental and tissue-specific manner. This indicates that the vimentin-derived segment of the fusion gene contains all the regulatory elements required for vimentin-specific expression. Immunohistochemical staining of fibroblast cultures derived from the transgenic mice with antibodies specific for vimentin and desmin demonstrated that the pVVim2 protein is assembled into filaments that co-localize with the endogenous vimentin filaments. The expression of pVVim2 protein in mesenchymal cells does not interfere with the function of vimentin in these cells. PMID- 3042383 TI - Divergent mRNA transcription in the chloroplast psbB operon. AB - The genes psbB, psbH, petB and petD for the components in photosystem II and the cytochrome b6/f complex are clustered and co-transcribed in liverwort Marchantia polymorpha chloroplasts. On the opposite DNA strand in the spacer region between the genes psbB and psbH, we deduced an open reading frame consisting of 43 sense codons, and designated it as the ORF43 gene. The ORF43 gene was actively transcribed in liverwort chloroplasts. The ORF43 transcripts were entirely complementary to a part of the primary transcripts of the psbB operon. Heterogeneous Northern hybridization showed that the mRNA transcripts for the ORF43 gene increased with greening in pea seedlings. This is the first demonstration of divergent overlapping transcription in chloroplasts. PMID- 3042387 TI - Diagnosis of pituitary tumors. AB - In the last two decades, significant strides have been made in the diagnosis and management of pituitary tumors. The identification, isolation, and characterization of the anterior pituitary hormones, the developments of sensitive and specific radioimmunoassays, and the significant advances in neuroradiologic, neurosurgical, and pathologic techniques have led to increasing recognition of pituitary tumors and their clinical syndromes. This article will review the current diagnostic approaches relating to prolactin, growth hormone-, thyrotropin-, and gonadotropin/alpha glycoprotein subunit-secretory, as well as nonsecretory, pituitary tumors. PMID- 3042385 TI - A carboxyl-terminal cysteine residue is required for palmitic acid binding and biological activity of the ras-related yeast YPT1 protein. AB - The Saccharomyces cerevisiae YPT1 gene codes for a ras-like, guanine nucleotide binding protein which is essential for cell viability. The functional significance of two consecutive cysteines at the very carboxyl-terminal end of this protein and in ypt homologues of other eukaryotic species was examined. YPT1 gene mutations were generated that either led to substitutions by serine or the deletion of one or both C-terminal cysteines. The consequences of the mutations were checked in cells after replacing the wild type with the mutant genes. It was found that as long as one of the cysteines was retained, the protein was fully functional. The YPT1 protein could be labelled with [3H]palmitic acid that appeared to be bound in an ester linkage. The wild-type protein was evenly distributed between soluble and membrane-associated proteins, the palmitoylated form was predominantly in the crude membrane fraction. The mutant protein lacking the C-terminal cysteines was not palmitoylated and was exclusively found in the soluble fraction. The extension by three residues, -Val-Leu-Ser, generating a ras typical C-terminal end, did not interfere with the mutant YPT1 protein's function although it resulted in a reduced labelling with palmitic acid. PMID- 3042388 TI - Disorders of antidiuretic hormone. AB - Disorders of thirst and vasopressin secretion present clinically in one of three ways: as hypotonic polyuria (DI), as hypodipsic hyponatremia, and as hyponatremia. In evaluating a patient with DI, the major challenge is to differentiate between primary polydipsia and neurogenic and nephrogenic DI. This is best accomplished through a series of steps that start with simple clinical observation, and progress, as necessary, to more complicated diagnostic procedures (Fig. 1). If the diagnosis is not clear from the clinical setting and the patient's history, the first step is to measure plasma osmolality and sodium under conditions of ad libitum fluid intake. If the results are clearly above the upper limit of normal range, primary polydipsia is excluded and the work-up can proceed directly to administration of vasopressin or DDAVP and/or a measurement of plasma vasopressin levels to differentiate between neurogenic and nephrogenic DI. If basal plasma osmolality and sodium fall within normal range, the standard dehydration test should be performed. If urine osmolality does not increase above that of plasma despite evident dehydration, primary polydipsia is excluded and the effect of vasopressin or DDAVP on urine osmolality should be examined to differentiate between neurogenic and nephrogenic DI. If administration of antidiuretic hormone increases urine osmolality by more than 50 per cent, the patient has severe neurogenic DI. If the increase in urine osmolality is less than 50 per cent, the patient has nephrogenic DI. In patients who do not concentrate urine above that of plasma in response to dehydration, the best approach is to measure plasma vasopressin, osmolality, and sodium after the latter have been increased above normal range by dehydration and/or infusion of hypertonic saline. When these results are plotted on a suitable nomogram (Fig. 2), neurogenic DI can be clearly diagnosed from the relative deficiency of vasopressin. In patients with normal vasopressin levels, primary polydipsia can be differentiated from nephrogenic DI by examining the relationship of urine osmolality to plasma vasopressin (Fig. 3), obtained during dehydration and/or graded vasopressin infusion. In evaluating a patient with sustained hypernatremia, it is only necessary to assess thirst, which can be done by a simple bedside observation. In a patient without obvious neurologic or cognitive impairment, absence of thirst in the face of plasma osmolality above 305 mosm/kg (plasma sodium above 150 mEq/L) is diagnostic for hypodipsic hypernatremia. In a patient who presents with hyponatremia, the main objective is to differentiate between hyper-, hypo-, and euvolemic (SIADH) types PMID- 3042386 TI - A gene which encodes a predicted protein kinase can restore some functions of the ras gene in fission yeast. AB - The ras1- mutation of the fission yeast Schizosaccharomyces pombe interferes with sexual differentiation by preventing conjugation and causing inefficient sporulation. From a gene library, we have isolated a gene, byr1+, which when in high copy number restores efficient sporulation to ras1- strains. byr1+ encodes a putative 340-amino acid protein product, the sequence of which strongly suggests that it functions as a protein kinase. Gene disruption experiments show that loss of byr1+ function does not interfere with mitotic growth but it completely prevents both conjugation and sporulation. byr1 is thus another important gene in the sexual differentiation pathway and we believe that at least part of ras1 function is to act directly or indirectly through byr1 to modulate protein phosphorylation. PMID- 3042389 TI - Evaluation of the infertile couple. AB - The evaluation of the infertile couple is usually a lengthy investigation in which all possible etiologic factors in both partners have to be considered. Optimal and cost-effective investigation requires adequate recognition of significant historical data and physical findings. Males without stigmata of endocrinopathies or general medical illnesses require an analysis of their semen as the minimum initial step of evaluation. Those suspected of deficient androgen production and/or action and those with abnormal sperm counts, motility, and/or morphology need assessment of their serum concentrations of selected reproductive hormones. When these initial investigations are negative and there are no demonstrable etiologic female factors underlying the state of infertility, specialized sperm function and sperm allergy testing needs to be performed. The initial investigation of the female partner is best served by assessing the frequency of ovulation and adequacy of corpus luteum function. Women without ovulatory defects should be assessed for the presence of the hostile cervical mucus and structural anomalies of the reproductive tract. Investigations of patients with menstrual dysfunctions should be based upon the presence or absence of hirsutism, changes in body weight, and evidence of other endocrinopathies or medical illnesses. Following the identification and normalization of causes of anovulation, further work-up of patients who remain infertile is similar to those with regular menstrual cycles. The diagnosis of idiopathic infertility is essentially by exclusion of all other causes. Algorithms for the diagnostic evaluation of most infertile couples are provided. PMID- 3042390 TI - Disorders of gonadal differentiation and congenital adrenal hyperplasia. AB - Ambiguity of the external genitalia may be based on disordered steroid biosynthesis or may arise from fundamental genetic and chromosomal abnormalities that produce failure of gonadogenesis. This article first discusses congenital adrenal hyperplasia and then considers the diverse abnormalities that cause disordered gonadal differentiation. PMID- 3042391 TI - Primary aldosteronism. Diagnostic evaluation. AB - The regulation of mineralocorticoid secretion and the pathophysiology of primary aldosteronism are reviewed. For conceptual and practical purposes, the diagnostic evaluation of primary aldosteronism is discussed as two series of studies. The first series involves the studies necessary to confirm the diagnosis. The second series of studies guides the therapeutic approach by distinguishing unilateral from bilateral adrenal disease. PMID- 3042392 TI - Diagnostic evaluation of pheochromocytoma. AB - Progress in the diagnostic evaluation of pheochromocytoma has taken place in biochemical studies and localization techniques. Measurement of fractionated catecholamines and their metabolites has been subjected to sensitivity and specificity assessment. Magnetic resonance imaging and isotopic imaging have led to much better localization of extra-adrenal tumors. Flow cytometry of the DNA of the tumor cells very likely indicates the malignant nature of the tumor. PMID- 3042393 TI - Carcinoid syndrome and disorders of systemic mast-cell activation including systemic mastocytosis. AB - Diagnostic approaches, clinical characteristics, and (briefly) therapy of the carcinoid syndrome and disorders of systemic mast cell activation are outlined. Mediators responsible for the humoral manifestations of the two syndromes are discussed. Because some of the clinical features of both disorders are similar (in particular, flushing), specific attention is given to the clinical differentiation of these disorders. PMID- 3042394 TI - Nucleotide and thioredoxin specificity of the manganese ribonucleotide reductase from Brevibacterium ammoniagenes. AB - The manganese-containing ribonucleotide reductase previously identified in gram positive bacteria has been purified and its nucleotide specificity and other requirements were determined. The enzyme isolated from Brevibacterium ammoniagenes is a ribonucleoside-diphosphate reductase which, in the presence of allosteric effectors, reduces all four common substrates at comparable rates; very little activity is observed in the absence of effector nucleotides. Ribonucleoside triphosphates are reduced at 20% the rate of the diphosphates. Cytidine and uridine nucleotide reduction is specifically stimulated by ATP and dATP, adenylate reduction by dGTP, and guanosine nucleotide reduction by dTTP. Unlike the iron-containing ribonucleotide reductase systems, high concentrations of dATP do not inhibit substrate reduction. The new bacterial enzyme tolerates high salt concentrations (up to 250 mM ionic strength) and does not require divalent metal ions for activity in vitro. The presence of thioredoxin has been demonstrated in heat- and acid-treated protein extracts of B. ammoniagenes and the protein was purified to homogeneity. It is very similar to the thioredoxins isolated from other organisms in relative molecular mass (12,000), isoelectric point (4.3) and enzyme-activating properties. In the presence of 0.3 mM dithiothreitol, the bacterial thioredoxin can serve as hydrogen donor for B. ammoniagenes ribonucleotide reductase in vitro, indicating the presence of a functional ribonucleotide reductase-thioredoxin system in these bacteria. The properties described in this and in our preceding paper in this journal [Eur. J. Biochem. 170, 603-611 (1988)] suggest that the B. ammoniagenes ribonucleotide reductase is intermediate in structure and specificity between the deoxyadenosylcobalamin-dependent and the iron-containing enzyme classes and that it is adapted to the specific requirements of deoxyribonucleotide synthesis in this organism. PMID- 3042395 TI - Dissociation of the sodium-ion-translocating oxaloacetate decarboxylase of Klebsiella pneumoniae and reconstitution of the active complex from the isolated subunits. AB - Oxaloacetate decarboxylase was reconstituted from the purified alpha subunit and a Triton X-100 extract of bacterial membranes devoid of this protein. Upon freezing of oxaloacetate decarboxylase in salt solutions, the enzyme was split into subunits and the catalytic activity was abolished. The catalytically active decarboxylase complex was reconstituted by decreasing the salt concentration of the dissociated sample. The conditions for the inactivation were critical for an optimum recovery of catalytically active enzyme during reconstitution, and modest dissociating conditions generally improved the yield of the reconstitutively active decarboxylase. The dissociated enzyme has been separated by chromatography on avidin-Sepharose into two fractions: fraction I, that was not retained on the column, consisted of the beta + gamma subunits, and fraction II consisted of the biotin-containing alpha subunit. Oxaloacetate decarboxylase was reconstituted from a mixture of the isolated alpha and beta + gamma subunits. The Na+ transport activity was recovered, if a mixture of subunits alpha and beta + gamma was incorporated into liposomes, or by a sequential reconstitution, starting with the formation of proteoliposomes with the integral membrane proteins beta + gamma and completed by an attachment of the peripheral subunit alpha. PMID- 3042396 TI - A protein covalently bound to ColE1 DNA. AB - ColE1 DNA was isolated from Escherichia coli as a relaxation complex of supercoiled DNA and proteins. Treatment of the complex with either protein denaturing agents (SDS, phenol etc.) or proteolytic enzymes converted the supercoiled DNA to an open-circular form (relaxation). The relaxation complex was separately labelled in vivo with [3H]Leu or [14C]Leu, [35S]Met or (32P)phosphate and extensively purified. Complete hydrolysis of the relaxed complex with DNase I and P1 nuclease produced a 36-kDa protein which, we believe, is covalently bound to ColE1 DNA. On the other hand, the relaxed complex was treated with tosylphenylalanylchloromethane-treated-trypsin and the DNA-peptide(s) produced was (were) isolated and digested with the nucleases as above. The resulting nucleotidylpeptide(s) was (were) isolated by DEAE-Sephadex chromatography. The only 5'-dCMP was released from the nucleotidylpeptide(s) by snake venom phosphodiesterase treatment. O-Phosphoserine was found in acid hydrolysates of the DNA-peptide(s). We suggest that in the relaxation event the 36-kDa protein becomes covalently linked to ColE1 DNA via a phosphodiester bond between dC and the serine residue. PMID- 3042397 TI - Import of proteins into mitochondria: a multi-step process. AB - Translocation of precursor proteins from the cytosol into mitochondria is a multi step process. The generation of translocation intermediates, i.e. the reversible accumulation of precursors at distinct stages of their import pathway into mitochondria ('translocation arrest'), has allowed the experimental characterization of distinct functional steps of protein import. These steps include: ATP-dependent unfolding of precursors; specific recognition of precursors by distinct receptors on the mitochondrial surface; interaction of precursors; specific recognition of precursors by distinct receptors on the mitochondrial surface; interaction of precursors with a general insertion protein ('GIP') in the outer mitochondrial membrane; membrane-potential-dependent translocation into the inner membrane at contact sites between both membranes; proteolytic processing of precursors; and intramitochondrial sorting of precursors via the matrix space ('conservative sorting'). The functional characteristics unveiled by studying mitochondrial protein import appear to be of general interest for investigations on intracellular protein sorting. PMID- 3042398 TI - Effect of nifedipine on arrhythmias in the acute phase of myocardial infarction. AB - In a double-blind placebo-controlled trial to study the effect of nifedipine on ventricular arrhythmias among patients with acute myocardial infarction, 434 patients with suspected myocardial infarction were randomized within 6 h from the onset of chest pain to treatment with nifedipine (p = 217) or placebo (p = 217). During the 48-h treatment period, a 10-mg capsule containing active drug or placebo was administered sublingually every 4 h for 24 h, then orally every 4 h for the next 24 h. Acute myocardial infarction was confirmed in 295 patients (146 in the nifedipine group and 149 in the placebo group). Twenty-four hour ECG tape analysis during 1-5 h from onset of chest pain showed that there was no significant difference in the number of patients with ventricular ectopics, ventricular couplets, ventricular tachycardia (3-9 beats), self terminating or sustained ventricular tachycardia between the two treatment groups. Also during the greater than 5-24 h from onset of chest pain, the numbers of patients with ventricular ectopics, multifocal, bigeminal or couplets, self-terminating ventricular tachycardia or sustained ventricular tachycardia did not differ significantly. However, there was a significant reduction in the number of patients with short runs of ventricular tachycardia (3-9 beats) in the nifedipine treated group. There was no significant difference among patients with ventricular fibrillation between the two treatment groups. PMID- 3042400 TI - Coronary artery spasm in a transplant patient. AB - The mechanism of coronary spasm is poorly understood. We report a case of coronary artery spasm in a cardiac transplant patient. Pharmacologic testing indicated denervation of the patient's heart. Therefore coronary artery spasm may occur despite cardiac denervation. This case demonstrates that intact autonomic nervous system and cardiac innervation are not essential for coronary artery spasm. PMID- 3042399 TI - Detection of myocarditis during the first year after discovery of a dilated cardiomyopathy by endomyocardial biopsy and gallium-67 myocardial scintigraphy: prospective multicentre French study of 91 patients. AB - The purpose of this study was to assess the frequency of inflammatory lesions in the myocardium of subjects with dilated cardiomyopathy and to determine if there was any correlation between the results of two methods of evaluation, one (endomyocardial biopsy) invasive and the other (gallium-67 scintigraphy) noninvasive. Of 115 subjects recruited in seven centres, 91 met the inclusion criteria (left ventricular dilatation greater than or equal to 100 ml m-2 and ejection fraction less than 55% with normal coronary arteriography) and had endomyocardial biopsy (mean five specimens) and Ga-67 myocardial scintigraphy after several days. Scanning was considered doubtful 19 times and positive 13 times. The histologic count of mononuclear cells in the myocardial interstitium in 20 fields was greater than 5 cells field-1 in only four cases. No correlation was found between the two methods. Subjectivity in the choice of the criterion of positivity of Ga-67 scintigraphy and difficulties in identifying lymphocytes upon pathological examination were the major problems encountered. Despite limitations, both techniques suggest that cellular infiltrates are minimal and quite infrequent in dilated cardiomyopathy. PMID- 3042401 TI - May non-invasive methods replace catheterization in quantification of aortic stenosis? PMID- 3042402 TI - Selection of a prosthesis for aortic valve replacement. PMID- 3042403 TI - Ventricular arrhythmias, syncope and sudden death in aortic stenosis. PMID- 3042404 TI - The natural history of aortic valve stenosis. AB - Despite different aetiologies, acquired aortic stenosis is a self-maintaining, slowly progressive process with good long-term prognosis. In 142 patients with mild stenosis, there was clinical progression within 10 years of the initial diagnosis in only 12% of patients. Twenty-five years after the diagnosis had been established, the severity of aortic stenosis was clinically unchanged in 38%, while 25% of patients had moderate stenosis and 35% had undergone valve replacement. Progression of moderate aortic stenosis was more rapid: the average time interval between the manifestation of moderate aortic stenosis and surgery was 13.4 years. Age at the onset of initial symptoms was related to aetiology: 39 +/- 18 years with rheumatic aortic stenoses, 48 +/- 6 years in patients with bicuspid valves who had no history of rheumatic fever, infective endocarditis or myocarditis, and 66 +/- 12 years in degenerative, calcific stenoses of tricuspid aortic valves. Patients with haemodynamically severe stenosis who had refused the recommended operation (n = 55) had an overall poor prognosis: mean survival averaged 23 +/- 5 months and the five-year probability of survival was 18 +/- 7%. All these patients died within 12 years of observation. Mean survival after the occurrence of angina pectoris was 45 +/- 13 months, after syncope 27 +/- 15 months, and after first occurrence of left heart failure 11 +/- 10 months. PMID- 3042406 TI - Valvular aortic stenosis and coronary atherosclerosis: pathophysiology and clinical consequences. AB - New methods of investigation (ECG, echocardiography, angiography, histology) allow a better understanding of the pathophysiology of valvular aortic stenosis (VAS) associated with coronary atherosclerosis. The progressive decrease in valvular aortic area modifies the coronary blood flow and leads to myocardial ventricular hypertrophy. These two mechanisms worsen left ventricular function. A significant atherosclerotic stenosis on a large coronary artery creates a considerable reduction of the available coronary blood flow. This reduction is permanent: present at rest, it is obviously increased during exercise. The study of the relationship between the severity of the VAS and the myocardial hypertrophy (MH) is of great interest. It seems that in VAS with coronary artery disease, different situations exist: (i) when the hypertrophy is severe (left ventricular mass greater than 180 g m-2), the angina pectoris is more attributable to the VAS than to the coronary lesions. Thus the removal of the aortic outflow obstruction is the most essential therapy; (ii) when the hypertrophy is less severe (left ventricular mass less than 180 g m-2), surgical treatment of the valvular lesion and myocardial revascularization are justified. PMID- 3042405 TI - Valvular aortic stenosis and asymmetric septal hypertrophy: diagnostic considerations and clinical and therapeutic implications. PMID- 3042407 TI - Coronary artery bypass graft surgery: late survival and follow-up. PMID- 3042408 TI - Ventricular arrhythmias in normal individuals and athletes. PMID- 3042409 TI - Exercise-induced arrhythmia. PMID- 3042410 TI - The implications of antiarrhythmic therapy in aviators. PMID- 3042411 TI - Implanted devices and aviation. PMID- 3042413 TI - Thromboembolic disease. AB - (1) Thromboembolic disease is a rare condition in physically fit subjects and is, therefore, likely to occur only rarely in aircrew. (2) The clinical diagnosis of both pulmonary embolism and deep venous thrombosis is unreliable. As the occurrence of these conditions has serious implications so far as the future licensing of airmen is concerned, it is of paramount importance to establish the diagnosis beyond doubt. (3) The chance of recurrence after an acute pulmonary embolus is low, particularly in those subjects in whom a definite acute and transient predisposing factor such as recent surgery or trauma can be identified. (4) When there is no definite predisposing factor to an acute pulmonary embolism, careful assessment is needed. The possibility that the embolus was the first sign of some serious sytemic illness, such as malignancy, should be considered and the possibility of rare clotting abnormalities sought. (5) Patients with subacute pulmonary embolism should be regarded as for those with acute pulmonary embolism and no obvious risk factor. A large proportion of these patients will have no predisposing factor and will require long-term oral anticoagulants as part of their routine therapy. Fortunately this condition is very rare. (6) Patients with repeated deep venous thrombosis, usually associated with chronic damage to the venous system of the leg, also may require long-term oral anticoagulants and their condition is likely to be exacerbated by long periods of inactivity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3042412 TI - Prolongation of the QT interval as an indicator of risk of a cardiac event. AB - The thesis (stated or implied) that acquired QT interval (or QTc) prolongation reflects physiological disturbance similar to those found in the congential syndromes has been assumed to be of central importance in many studies of QT prolongation in the clinical setting. There is no evidence to support this assumption. It is suggested that the use of rate correction of the measured QT interval has obscured our understanding of repolarization abnormalities. Although a QT interval corrected for heart rate may reflect a relationship between the duration of repolarization and heart rate in a single value (e.g. QTc) the corrected value clearly embodies complex influences other than those directly due to rate and for which no 'correction' is, as yet, possible. PMID- 3042415 TI - Obliterative and restrictive cardiomyopathies. PMID- 3042414 TI - Long-term morbidity and mortality after aortic and mitral valve replacement with tissue valves and certification to fly. PMID- 3042416 TI - Sarcoidosis of the heart. AB - Patients who have bilateral hilar lymphadenopathy (BHL) alone need annual follow up with chest X-ray, ECG, 24-h monitoring, exercise testing, lung function and possible thallium 201 scanning. Provided these tests are normal or negative, there is no evidence of other organ or parenchymal lung involvement and the extent of BHL is stable or regressing, then restriction to multicrew operation should be required until regression has occurred or a period of two years has elapsed. Although a patient with generalized or systemic sarcoidosis and no apparent cardiac involvement should be allowed to drive a car, certification to fly should be dependent on physical well being and restricted to multicrew operation. With our present inability to diagnose cardiac involvement in sarcoidosis with complete sensitivity, this restriction should be indefinite as cardiac involvement in sarcoidosis may only become apparent many years after the diagnosis of systemic sarcoidosis has been made. Patients with known cardiac involvement in sarcoidosis should also be refused a driving licence unless they are under regular and close supervision for possible arrhythmic complications. Permanent pacing may be needed. Patients with known cardiac sarcoid should be refused certification to fly. PMID- 3042417 TI - Post-viral pericarditis. PMID- 3042418 TI - Non-pharmacological antihypertensive therapy. PMID- 3042419 TI - Newer anti-hypertensive agents and their use by aircrew. PMID- 3042421 TI - Peripheral vascular disease as a risk factor for ischaemic heart disease. PMID- 3042420 TI - Evaluation of the cerebral effects of antihypertensive medication. PMID- 3042422 TI - Ischaemic heart disease. Critical review of the usefulness of non-invasive investigation of asymptomatic individuals. PMID- 3042423 TI - Risk stratification after myocardial infarction with and without coronary artery bypass grafting. PMID- 3042424 TI - The relationship between Ga-67 accumulation and cell cycle in malignant tumor cells in vitro. AB - Previously we reported that the deposition of 67Ga into malignant tumor may be a sensitive index of proliferative activity in tumor cells. For the purpose of elucidation of this hypothesis, we investigated the relationship between the accumulation of 67Ga into malignant tumor cells and the cell cycle in vitro. We discovered that the uptake of both 67Ga and 59Fe into synchronized mouse tumor cells reaches a peak at the G2 stage which precedes cellular proliferation. Both iron and transferrin are specifically required by cells in culture for cell division, and the fact that 67Ga and 59Fe uptake into tumor cells peaks at the G2 stage of the cell cycle suggests that there is a correlation between 67Ga uptake and the rate of cellular proliferation in malignant tumor cells. PMID- 3042425 TI - Transplantation of isolated hepatocytes into the pancreas. AB - In previous research into hepatocyte transplantation (HTX) the spleen was the preferred acceptor organ for isolated donor hepatocytes. In this study the pancreas was tested as an acceptor organ for HTX. HTX into the pancreas or spleen was performed by injection of 10(7) isolated hepatocytes into the parenchyma of these organs. Intrapancreatic hepatocytes showed good viability 3 months after syngenic HTX as assessed by histological and immunocytochemical parameters. Definite proof of sustained metabolic activity of normal hepatocytes, 3 months after transplantation into the pancreas of congenitally jaundiced rats, was obtained by demonstration of bilirubin conjugates in bile of the recipients: 4.0% of total biliary bilirubin was conjugated. Intrasplenic HTX, however, was more effective and resulted in a conjugated fraction of 17.7% of total biliary bilirubin (p less than 0.001). Reduction of total plasma bilirubin was significant with both methods, but more pronounced in intra-splenic HTX. Bile drainage from the hepatocellular transplant via the pancreatic excretory system into the gut was not observed: conjugated bilirubins were not found in pancreatic juice of HTX-treated jaundiced rats. Intrapancreatic HTX did not adversely affect the host rat; evidence of pancreatitis or diabetes was not found. It is concluded that the pancreas is a suitable acceptor organ for HTX. However, intrapancreatic HTX appears to be less effective than intrasplenic HTX in the treatment of enzyme deficiency disease. PMID- 3042427 TI - Experimental model of heterotopic cardiac transplantation for evaluation of graft viability and function. AB - An experimental model of heterotopic intrathoracic cardiac transplantation making possible an evaluation of graft viability and function has been studied. The technique requires only two anastomoses in the normothermic state without cardiopulmonary bypass. In this model, the ascending aorta of the recipient animal may be occluded completely, enabling an increasing preload on the graft. In case of adequate myocardial preservation of the cardiac graft, the aorta may be completely occluded and the recipient systemic circulation can be sustained by the graft. However, in case of inadequate graft myocardial function, or in case of size mismatch, the recipient heart alone contributes to the circulation. This technique appears quite useful because of its simplicity, the elimination of the need for extracorporeal circulation while allowing a functional evaluation following cardiac graft preservation. PMID- 3042426 TI - A computer-assisted device for the intraoperative CT-correlated localization of brain tumors. AB - In a series of 72 open brain tumor operations the 32P (radiophosphorus) test proved to be valuable for the removal of visually not detectable tumor residues. As during resection a topographical orientation in the depth of tumor cavities was nearly impossible, a computerized measuring device was developed which enables the surgeon to locate the tumor boundaries by simultaneous comparison with preoperative CT scans. For 32P-beta emission measurements a miniature semiconductor probe can be attached to the tip of the four-axis digitizing arm allowing the evaluation of radicality by topographic distinction between tumor ramifications and normal brain tissue. The device was tested during 3 craniotomies and the perspectives of computerized stereotaxy are discussed. PMID- 3042428 TI - Indications of chlormethiazole as a protective agent in experimental endotoxemia. AB - Chlormethiazole, which is derived from the thiazole moiety of thiamine, possesses sedative, hypnotic and anticonvulsant properties. This anesthetic agent was compared with ketamine in a porcine model of endotoxemia to evaluate effects on cardiovascular and pulmonary function, oxygen delivery and survival. Continuous 6 hour intravenous infusion of Escherichia coli endotoxin caused pronounced pulmonary and cardiovascular derangement and decreases in oxygen delivery and pH in 13 pigs given ketamine anesthesia. Eight of thirteen pigs survived the observation period. Contrastingly, 10 pigs given chlormethiazole anesthesia and endotoxin showed a significantly attenuated response. Thus, the increases in mean pulmonary arterial pressure, venous admixture and extravascular lung water were significantly lower and the decreases in cardiac output, oxygen delivery and pH were significantly modified by chlormethiazole. All 10 pigs survived the observation period. Chlormethiazole may increase the clearance of endotoxin and thus ameliorate the endotoxin response. Although extrapolating from animal data requires great caution, these data may favor the use of chlormethiazole in septic states requiring surgical intervention and anesthesia and as sedation in critically ill septic patients in our intensive care units. PMID- 3042429 TI - Changes in granulocyte chemiluminescence and plasma fibronectin concentrations following major blunt trauma. AB - Chemiluminescence activity of granulocytes in phagocytosis of zymosan and Escherichia coli and their responses to chemoattractant N-formylmethionyl-leucyl phenylalanine (FMLP) were evaluated in 13 major blunt trauma patients (Injury Severity Score 31 +/- 6) and their plasma fibronectin concentrations were measured. Chemiluminescence responses to zymosan and E. coli were at control levels immediately after injury and a week thereafter, but responses to FMLP were increased compared to the controls (p less than 0.05). Plasma fibronectin concentrations were depressed on the day after trauma (p less than 0.001) but increased to control values over 1 week. The changes had no correlation with the patients' recovery. PMID- 3042430 TI - Long-term effects of guar gum and microcrystalline cellulose on glycaemic control and serum lipids in type 2 diabetes. AB - The effects of guar gum (GG) and microcrystalline cellulose (MC) on metabolic control and serum lipids were compared in a double-blind, cross-over trial in 18 poorly controlled Type 2 diabetic patients. There were two 12 week treatment periods separated by a 4 week wash-out period. A significant reduction in fasting BG was found after 6 weeks treatment with GG, but the initial level was regained after further 6 weeks, at the end of the treatment period. No statistically significant change in fasting BG was observed with MC. Serum cholesterol was lowered by 10% during GG treatment. Microcrystalline cellulose had no effect on serum lipids. The results suggest, that during 12 weeks supplementation with guar gum, the improvement in glycemic control was not sustained, but that it might reduce the risk of macrovascular disease in diabetic patients. PMID- 3042431 TI - Identification of individual tumor necrosis factor/cachectin-producing cells after lipopolysaccharide induction. AB - A method to detect single tumor necrosis factor alpha (TNF alpha)-producing cells, based on immunofluorescence staining, is presented. We have developed a rapid and simple procedure to permeabilize blood leukocytes for antibodies without causing cell aggregation. Using monoclonal as well as polyclonal TNF alpha-specific antibodies cytoplasmic TNF alpha could be detected in lipopolysaccharide (LPS)-stimulated monocytes in identical ways. In addition, one out of 15 tested fresh blood samples from healthy blood donors contained occasional TNF alpha-producing cells. We found a well-defined staining of the intracellular TNF alpha with a local, polar accumulation in a juxtanuclear position of the cell. This finding most probably indicated the presence of the monokine in the Golgi organelle because the sequential staining for TNF alpha and the Golgi zone by specific antibodies coincided. Thus, we believe the TNF alpha accumulation at this site reflects the production rather than cellular uptake of the cytokine, since the incubation of blood leukocytes with recombinant TNF alpha did not lead to any detectable staining. By performing two-color staining of cell surface antigens and cytoplasmic TNF alpha of LPS-stimulated blood leukocytes we could demonstrate that monocytes exclusively contributed to the TNF alpha synthesis. At the peak of the response, which occurred 2-3 h after LPS exposure, 50-60% of the blood monocytes produced TNF alpha. We noted a rapid decline in the number of TNF alpha-producing cells already 6 h after initiation of these cultures. PMID- 3042432 TI - Evidence for diabetogenic action of bumetanide in mice. AB - The effect of bumetanide on carbohydrate metabolism was studied in mice. Intraperitoneal injection of 50 or 100 mg bumetanide/kg body weight resulted in an acute and transient hyperglycaemia. Pretreatment with 240 mg probenecid/kg body weight reduced the diuretic effect but potentiated the hyperglycaemic effect of bumetanide (50 mg/kg body weight). The glucose tolerance was impaired, and there was an elevated serum glucose and glucose/insulin ratio 2 h after a single injection of bumetanide (100 mg/kg body weight). It is suggested that bumetanide has an acute effect on carbohydrate metabolism in mice that is not secondary to diuresis and that the reduced glucose tolerance may, at least in part, be due to a reduced capacity to secrete insulin. PMID- 3042433 TI - The use of mathematical models in the epidemiological study of infectious diseases and in the design of mass immunization programmes. PMID- 3042435 TI - Antigenic analysis of Haemophilus ducreyi by Western blotting. AB - Twenty-one strains of Haemophilus ducreyi were analysed by Western blotting using two antisera produced in mice. Common antigens of molecular weights 58, 46, 41, 28, 22 and 16 kDa were detected in all the strains. The antigens were protein in nature, since they could not be detected in whole-cell lysates which had been treated with proteinase K. The H. ducreyi strains showed antigenic cross reactivity with strains of H. influenzae and H. parainfluenzae, but showed minimal or no cross-reactivity with seven other species of bacteria. PMID- 3042434 TI - Serratia marcescens infection associated with early abortion in cows and buffaloes. AB - Serratia marcescens was isolated in pure culture from cases of septic abortion in 4 cows on one farm and 10 buffaloes on two other farms. A reddish vaginal discharge was observed after abortion in all animals and in the internal organs of the aborted fetuses. All but two of the isolates produced prodigiosin, and two of the isolates from buffaloes were atypical in that they fermented raffinose. O serological, bacteriophage and bacteriocin typing revealed four different strains. All cows were infected by the same strain, and this strain was also isolated from the semen of a breeding bull on the same farm. In another farm a strain of serotype O 14 was isolated from 6 of 10 buffaloes, and two other distinct strains were isolated from the remainder. The strain from the cattle was sensitive to gentamicin and so were two of the buffalo isolates. The infected cows were treated with intra-uterine gentamicin and the organism disappeared from cervical mucus after 3 days. Each animal after abortion showed a raised titre of agglutinating antibody to their respective isolate. A survey of 1172 healthy buffaloes and cattle gave an incidence of 1.8% with raised titres towards S. marcescens. PMID- 3042436 TI - Serological tests in leprosy. The sensitivity, specificity and predictive value of ELISA tests based on phenolic glycolipid antigens, and the implications for their use in epidemiological studies. AB - This paper examines the sensitivity and specificity of two ELISA assays for IgM antibodies to Mycobacterium leprae, one employing natural phenolic glycolipid and the other employing a synthetic disaccharide glycoconjugate as antigen. Estimates of sensitivity and specificity are derived, based on a panel of sera from leprosy cases in Malawi and various non-leprosy controls from the UK. Though both assays were able to identify a high proportion of multibacillary patients, neither was able to detect a high proportion of paucibacillary patients without considerable loss of specificity. The implications of the inverse relationship between sensitivity and specificity are discussed with reference to the predictive value of such tests in such areas as Malawi, where the large majority of cases are paucibacillary. PMID- 3042437 TI - Carriage of Neisseria meningitidis: investigations in a military establishment. AB - A prevalence study of personnel on a Royal Naval Air Station revealed that 23.0% of 2479 personnel were carrying a meningococcus. Selected groups of personnel were subsequently swabbed monthly for a year. We have shown that it is only by repeated swabbing and the use of optimal methods including enrichment media that one can have a hope of identifying the 'true' carriage rate. A presumed virulent strain of Neisseria meningitidis B15 P1.16 was repeatedly isolated from three personnel who remained well, as did their colleagues both at their work place and socially. The study served to emphasize our lack of knowledge of the virulence factors associated with N. meningitidis. PMID- 3042438 TI - The use of alkalinity and incubation at 9 degrees C for improved recovery of Yersinia spp. from faeces. AB - Recovery of Yersinia enterocolitica and related strains from faecal samples enriched in 1% buffered peptone water (pH 7.2) and incubated at 4 degrees C for 7 21 days was compared with recovery from 1% peptone water buffered to pH 8.0 and incubated over the temperature range 4-26 degrees C. Best recovery was obtained by use of the alkaline medium incubated at 9 degrees C. Greatest recovery was obtained after incubation for 10-14 days, but most strains (greater than 75%) were recovered after 1 week. PMID- 3042439 TI - Identification of encapsulated and non-encapsulated Yersinia pestis by immunofluorescence tests using polyclonal and monoclonal antibodies. AB - Rabbit polyclonal hyperimmune antibodies to Yersinia pestis, and a mouse monoclonal antibody against the capsular antigen fraction 1 (F1) were compared in immunofluorescence (IF) tests. Fluorescent antibody conjugates were prepared from polyclonal antisera to four F1 positive Y. pestis strains; the conjugated antibody to strain A1122 gave the strongest IF staining of F1 positive and F1 negative Y. pestis strains. Indirect assays were rejected in favour of direct assays utilizing polyclonal and monoclonal reagents because the increased background staining reduced the effective contrast of bacterial visualisation. Polyclonal conjugates gave fairly homogeneous staining of Y. pestis bacterial populations, but in monoclonal assays a skew distribution of fluorescence intensity was observed, the majority of bacteria being poorly stained. The proportion of cells stained well by the monoclonal sufficed for easy identification of Y. pestis of the F1 positive phenotype however, and staining was not affected by washing the bacteria or treating them with formaldehyde. Y. pestis strains of the F1 positive genotype reacted with the monoclonal if bacteria were grown at 37 degrees C but not if the growth temperature was reduced to 25 degrees C thus preventing capsule production. The polyclonal conjugate reacted with bacteria of these strains that had been grown at either temperature. Strains of F1 negative genotype grown at either temperature reacted with the polyclonal conjugate but not with the monoclonal. Cross reactions between the polyclonal reagents and Y. enterocolitica biovar 2, serovar O 8 could not be removed by selective absorption; however, the monoclonal antibody gave no cross reaction. The F1 phenotypic status of bacterial preparations was verified by ELISA measurement of the fraction 1 antigen concentration. Antigen levels for F1 positive and F1 negative phenotypes differed by about three logs for suspensions of Y. pestis harvested from solid media. The polyclonal and monoclonal direct IF tests applied to spleen and blood smears of laboratory mice infected with Y. pestis were able to differentiate between lethal infection with an F1 positive strain carrying all four classical virulence determinants, an F1 positive vaccine strain, and an F1 negative strain. PMID- 3042440 TI - Salmonella typhimurium phage type 141 infections in Sheffield during 1984 and 1985: association with hens' eggs. AB - Food poisoning due to Salmonella typhimurium phage type 141 was unusual in the Sheffield area before 1984. The sudden increase in incidence of this phage type during 1984 and 1985, and its causative role in several small outbreaks in this period have been investigated. Epidemiological and laboratory investigations suggested that hens' eggs were the most likely source of S. typhimurium phage type 141. PMID- 3042441 TI - First recognized community outbreak of haemorrhagic colitis due to verotoxin producing Escherichia coli O 157.H7 in the UK. AB - The first recognized outbreak of haemorrhagic colitis due to Escherichia coli O 157.H7 in the United Kingdom affected at least 24 persons living in East Anglia over a 2-week period. The illnesses were characterized by severe abdominal pain and bloody diarrhoea of short duration. Eleven patients were admitted to hospital and there was one death. Patients were mainly adult women who had not eaten out of the home in the 2 weeks before onset. Unlike previously reported outbreaks hamburgers were not the vehicle of infection, and a case-control study suggested that handling vegetables, and particularly potatoes, was the important risk factor. PMID- 3042442 TI - Granulocyte-macrophage colony-stimulating factor regulation in murine T cells and its relation to cyclosporin A. AB - The coordinated expression of the hematopoietic growth factors, interleukin 3 (IL3) and granulocyte-macrophage colony-stimulating factor (GM-CSF) in activated T cells suggests common synthetic pathways. However, cyclosporin A (CsA) appears to differentially effect the synthesis of these two lymphokines. When the supernatants from the concanavalin A-stimulated thymoma EL-4 or the T-cell hybrid 2B4 were assayed on the GM-CSF/IL3-dependent PT-18 and on the IL3-dependent DA-1 cell lines, IL3 activity could not be detected following CsA treatment, but substantial growth activity, which was identified as GM-CSF, was observed on the PT-18 cell line. CsA did not affect the kinetics of GM-CSF release, but inhibited the release of IL3 over a period of 40 h after the cells were stimulated and treated with CsA. In addition, CsA could be added up to 1 h after stimulation of the cells without affecting GM-CSF activity but inhibiting completely the IL3 activity. To substantiate the inability of CsA to inhibit GM-CSF activity, GM-CSF gene expression was evaluated. By Northern analysis GM-CSF mRNA was not inhibited by doses of CsA up to 1 microgram/ml. The data reveal that CsA can dissociate between the production of IL3 and GM-CSF, suggesting that these two CSFs are regulated by different mechanisms. PMID- 3042443 TI - Transfusion of bone marrow red cells during bone marrow harvests. AB - Large volumes of bone marrow may be required for certain types of autologous bone marrow transplants. The present study was done to determine whether red cells obtained during a bone marrow harvest would be useful in reducing homologous transfusion requirements. A group of patients receiving standard transfusion support during the harvest (group 1) was compared to a group that received processed bone marrow red cells (PBMRBC) (group 2). Using the Cobe 2991 cell processor, 90% of the harvested bone marrow red cells were extracted and transfused during the procedure. Group 2 received a median of 1500 ml of blood processed from the bone marrow or 413 ml (volume of marrow processed x hematocrit) of red cells. Infusion of the PBMRBC reduced the homologous transfusion requirement from 6.5 units to 3.0 units (p = 0.02). In addition, group 1 had a 20% decrease in hematocrit following transfusion compared to the pre-harvest hematocrit, as opposed to an 8% decrease in group 2 (p = 0.02). This study indicates that PBMRBC can reduce the homologous transfusion requirements during an autologous bone marrow harvest. PMID- 3042444 TI - Characterization of natural killer cells cultured from human bone marrow cells. AB - Human bone marrow (BM) cells, depleted of nylon wool-adherent cells, T cells, and natural killer (NK) cells, were cultured in medium containing recombinant interleukin 2 (rIL2). After 21 or 24 days in culture, numerous lymphoid cells with multiple azurophilic granules and a morphology similar to large granular lymphocytes (LGL) were found. Two-color analysis of surface phenotype showed many of these cells to be NKH1-positive and a limited number of cells had other NK markers such as CD16, CD2, or CD8. The CD3 antigen was not coexpressed with NKH1. The cultured BM cells were cytotoxic for K562, Daudi, and Raji cell lines. The NKH1+, CD2-, CD3-, CD16- cells were sorted and, in addition to having the LGL morphology, were found to be cytotoxic for K562 cells (NK [K562]). The generation of NK(K562) activity was significantly suppressed by 5-bromodeoxyuridine plus ultraviolet light treatment, indicating that DNA synthesis is required. These experiments suggest that the described culture conditions allow differentiation of progenitor cells, into immature, but functionally active, NK cells. PMID- 3042446 TI - Risk of HIV infection on patients and staff of two dialysis centers: seroepidemiological findings and prevention trends. AB - In view of the present world-wide diffusion of HIV, we evaluated the possible presence of persons infected by HIV or suffering from AIDS among the patients and staff of two Dialysis Centers. In the past these centers have been found to be at a high risk for HBV infection. The results of this seroepidemiological study, though rather reassuring for the time being, have led the authors to propose specific prevention rules, owing to the nature of activities in these medical structures and in light of the fact that the presence of HIV is beginning to be reported in other such centers. PMID- 3042447 TI - Acute intestinal infections in Europe. A review of reported cases. AB - Acute intestinal infections still constitute one of the leading causes of death in the world and a major cause of morbidity in Europe. Unfortunately, most European countries do not have an information system oriented towards acute intestinal infection surveillance. The present work is an analysis of cases of typhoid fever, Salmonellosis, Shigellosis, foodborne infections, and acute gastroenteritis reported from January 1980 to December 1985. Data from 24 of the 33 countries belonging to WHO Regional Office for Europe (EURO-WHO) were considered. Data for 1980 and 1981 were taken from World Health Organization statistics, while data for 1982-85 were taken from bulletins sent to EURO-WHO and WHOCCHDS by individual countries. Data was entered in an IBM 4341 computer system and a data base was organized using a general purpose inquiry language (IBM's APLDI). Rates per 100,000 were calculated using as a denominator 1983 WHO official population figures. The quality of the data is a great problem, as the only available sources of information are official national reports which underestimate the actual incidence of diseases. For typhoid fever, Mediterranean countries show much higher incidences than the rest of Europe, although a decreasing trend can be seen for all European countries. We estimate that the data for typhoid fever, Salmonellosis and Shigellosis are reliable, while reports of foodborne infections and acute gastroenteritis represent only a very small percentage of the actual number of cases. PMID- 3042445 TI - The hemolysin of Escherichia coli. AB - Many strains of E. coli elaborate a hemolysin which is responsible for the zone of beta-hemolysis surrounding bacterial colonies on blood agar. The significance of this cytolysin as a determinant of bacterial pathogenicity has been established in animal models with the use of genetically engineered, isogenic bacterial strains. An analogous role in human infections has been inferred from the high association of hemolysin production with disease. Studies at a molecular genetical level have defined 4 genes that are required for the synthesis, post translational modification and secretion of the hemolysin. The structural gene hlyA encodes for a 107-110,000 polypeptide, which must be modified in an unknown manner to its active form by the product of the neighboring hlyC gene. Genes hlyB and hlyD encode for proteins that export the molecule to the extracellular medium. The signal for secretion is contained in the C-terminal portion of the toxin molecule. The secreted hemolysin attacks plasma membranes of target mammalian cells by inserting as a monomer into the bilayer and generating hydrophilic transmembrane pores of approximately 2 nm effective diameter. The pores display a marked selectivity for cations over anions and pore-opening is dependent on the presence of a correct transmembrane potential. Binding to a membrane target does not require the presence of a specific receptor, and pores may be generated in planar lipid membranes consisting solely of phosphatidylcholine. Pore formation in nucleated cells can trigger secondary reactions such as stimulation of arachidonate metabolism with release of lipid mediators, probably initiated by passive influx of extracellular Ca2+.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3042448 TI - Aspects of imported malaria at a district general hospital in non-endemic Kuwait, Arabian Gulf. AB - There is no indigenous mosquito-borne transmission of malaria in Kuwait. However, in a five year period at a district general hospital, the number of laboratory diagnosed cases of malaria increased annually from 25 to 84, a rise of 336%. Except for two induced infections, all were imported, mainly from the Indian subcontinent. Plasmodium vivax was responsible for 87.29% of the cases; P.falciparum (12.05%), a mixed infection of P.vivax and P.falciparum (0.33%) and a case of P.ovale (0.33%) were also identified. Rapid preparation of acetone fixed, Giemsa-stained thick blood films, a heightened awareness of the infection, examination of multiple samples of blood from patients and the general resurgence of malaria in endemic areas were some of the factors responsible for the high number of cases diagnosed. Most patients were young males and presented with clinical malaria due to P.vivax between May and October each year, an apparent seasonal peak. However, many were already resident in the country for a variable period. Patients with P.falciparum though, presented clinically within two weeks of arrival in the country. Parasite densities were calculated to monitor the progress of treatment and identify quickly any possible chloroquine-resistant P.falciparum strains. A policy of active prophylaxis is suggested to stem the tide of imported malaria. PMID- 3042449 TI - The Dienes effect as an epidemiological tool in a paraplegic unit. AB - The authors study the Dienes effect in 136 P.mirabilis and 2 P.vulgaris strains, isolated from different infection or colonization sites, in 27 spinal cord injured patients, admitted to a Paraplegic Unit, during a 9 month period. It is shown that 7 cross-infection and 18 cross-contamination episodes occurred, affecting 13 of 27 patients, which indicates the great spreading capacity of these microorganisms; 97% of the results obtained were observed once again after 6 months' storage of strains. In addition, in 7 of these strains their resistance to several antibiotics was cured by treating them with acriflavine salts, demonstrating that the Dienes effect persists even when resistance to antibiotics is modified; the authors conclude that the Dienes effect is an accurate stable epidemiological tool to identify cross-infections and its origins, and to facilitate the interruption of the chain of infection. PMID- 3042452 TI - The spleen and pooling of blood cells. PMID- 3042450 TI - Chlamydia specific IgG and IgA antibodies in women with obstructive infertility as determined by immunoblotting and immunoperoxidase assays. AB - The prevalence rate of IgG and IgA antibodies to Chlamydia was analyzed in 50 women with laparoscopy-verified tubal infertility and in 50 age-matched control women by single serovar (L2) inclusion immunoperoxidase assay (IPA) and by immunoblotting technique (IB). Women with tubal infertility had significantly (p less than 0.001) elevated IPA Chlamydia IgG antibody titer greater than or equal to 128 and greater than or equal to 256 than controls (64% vs 16%. Odds ratios = 9.3 and 50% vs 10%, Odds ratio = 9 respectively). The prevalence rate of IPA IgA antibody titer (greater than or equal to 16) to Chlamydia was also significantly higher (p less than 0.001) in women with tubal infertility than controls (48% vs 8%, Odds ratio = 10.6). Antibodies to at least 19 chlamydial structural polypeptides ranging in molecular weight from 30 kD to 204 kD, were detected by the IB technique in the IPA seropositive sera. Antibodies to 57-60 kD were detectable in almost all the IPA IgG and IgA seropositive sera. The prevalence rate of IgG antibody to 57 kD-60 kD was significantly higher in women with obstructive infertility than healthy woman (84% vs. 56% p less than 0.01; Odds ratio = 3.8). More significantly, higher differences to 57-60 kD polypeptide were found in the case of IgA between the infertile women and controls (52% vs. 10%, p less than 0.001; Odds ratio = 9.7). The significance of IPA and IB technique for screening of infertile women is discussed. PMID- 3042451 TI - Comparison among enterotoxigenic strains of Escherichia coli isolated in Italy and Somalia. AB - Nine strains of ETEC isolated in Italy have been compared with 13 isolates from Somalia with respect to toxin production, serotype and antimicrobial resistance pattern. None of the strains isolated from Italy belonged to any serotype or serogroups found among the strains from Somalia. Remarkable differences between the two groups of isolates were also observed with regard to the susceptibility to antimicrobials and the presence of R-plasmids. These findings suggest that ETEC strains isolated in Italy are not related to the strains widespread in Somalia and, generally, in developing countries. PMID- 3042453 TI - Multiple myeloma in central and northern Norway 1981-1982: a follow-up study of a randomized clinical trial of 5-drug combination therapy versus standard therapy. AB - In a randomized study of 92 previously untreated patients with multiple myeloma, the intention was to document the possible beneficial effect of combination chemotherapy including vincristine, carmustine, alkylating agents and prednisone, as compared to conventional therapy with melphalan and prednisone. Major prognostic factors did not differ significantly between the treatment groups. With the 2-drug therapy and 5-drug combination therapy, 48 and 54% of the patients achieved remission, respectively. Median survival for patients treated with the 2-drug regimen and 5-drug regimen was 29 and 33.5 months, respectively. No significant difference was found between the survival curves for stage III patients treated with the two regimens. After 12 months, patients who had achieved remission were randomized to have treatment discontinued or to have maintenance treatment. The numbers of relapses, remission duration and survival of the two groups were similar. PMID- 3042454 TI - Stop making sense: or Regulation at the level of termination in eukaryotic protein synthesis. AB - An increasing number of examples of translational regulation at the level of termination has been recently reported in eukaryotes. This paper reviews our present knowledge on this topic and proposes an understanding of these regulations by relating the study of viral gene expression to a comprehensive view of the mechanisms and components of the translational process. PMID- 3042455 TI - Tyrosine sulfation is not the last modification of entactin before its secretion from 3T3-L1 adipocytes. AB - Tyrosine sulfation of entactin was studied by labeling of 3T3-L1 adipocytes with [35S]methionine or H2 35SO4 in the presence or absence of tunicamycin or monensin. Four precursors (EN1-4) at different steps of modification were detected in addition to mature entactin. Under normal conditions, EN2 and mature entactin were intracellular species, and the latter was sulfated and secreted. Inhibition of co-translational transfer of N-linked oligosaccharides by tunicamycin produced EN1 and EN3 as intracellular species, and EN3 was sulfated and secreted. Interruption of protein transport from medial to trans (distal) Golgi cisternae by monensin, and consequent blockage of terminal glycosylation caused intracellular accumulation of EN4. EN4 was sulfated and of different size compared to mature entactin. These facts suggested that tyrosine sulfation of entactin occurs in medial Golgi cisternae and is not the last modification before its secretion. Our results appeared inconsistent with recent observations by Baeuerle and Huttner [(1987) J. Cell Biol. 105, 2655-2664] that tyrosine sulfation of IgM occurred within the trans (distal) Golgi cisternae as the last modification before its exit from the Golgi complex. PMID- 3042456 TI - Identification of a cysteine residue as the binding site for the dipyrromethane cofactor at the active site of Escherichia coli porphobilinogen deaminase. AB - The dipyrromethane cofactor of Escherichia coli porphobilinogen deaminase was specifically labelled with 13C by growth of the bacteria in the presence of 5 amino[5-13C]levulinic acid. Using 13C-NMR spectroscopy, the structure of the cofactor was confirmed as a dipyrromethane made up of two linked pyrrole rings each derived from porphobilinogen. The chemical shift data indicate that one of the pyrrole rings of the cofactor is covalently linked to the deaminase enzyme through a cysteine residue. Evidence from protein chemistry studies suggest that cysteine-242 is the covalent binding site for the cofactor. PMID- 3042457 TI - Complete amino acid sequence of protein B. AB - The complete amino acid sequence of protein B (= CAMP factor) of Streptococcus agalactiae has been determined. The sequence data were obtained mainly by manual sequencing of peptides derived from digestion with lysyl-peptidase, clostripain and Staphylococcus aureus protease and by solid phase sequencing of cyanogen bromide fragments. The protein contains 226 amino acids and has an Mr of 25,263. The sequence was compared with sequences of other Fc-binding proteins and partial sequence homology was found between protein B and the Fc-binding region of protein A. PMID- 3042458 TI - The amino-terminal sequence of the Xenopus laevis mitochondrial SSB is homologous to that of the Escherichia coli protein. AB - Two closely related forms of the single-stranded DNA binding protein purified from Xenopus laevis oocytes mitochondria have been identified. Their amino terminal sequences exhibit homology with the Escherichia coli SSB protein. PMID- 3042459 TI - Protein chemical characterization of Mucor pusillus aspartic proteinase. Amino acid sequence homology with the other aspartic proteinases, disulfide bond arrangement and site of carbohydrate attachment. AB - The amino acid sequence of Mucor pusillus aspartic proteinase was determined by analysis of fragments obtained from cleavage of the enzyme by CNBr and limited tryptic digestion. The proteinase is a single polypeptide chain protein containing 361 amino acid residues, cross-linked by two disulfide bonds. A sugar moiety composed of two GlcNAc residues and four neutral sugar residues is asparagine-linked to the chain. The sequence of M. pusillus proteinase is highly homologous with the M. miehei proteinase (83% identity). The homology with other aspartic proteinases is low (22-24%) and indicates that the Mucor proteinases diverged at an early evolutionary phase. The most conservative regions of the molecule are those involved in catalysis and forming the binding cleft and the core region of the molecule. PMID- 3042460 TI - An improved purification of lactose permease. AB - The integral membrane protein lactose permease of Escherichia coli was purified to homogeneity in detergent micelles. No other proteins could be detected in the final product. The molar ratio of lactose permease to lipid was less than 0.2 in detergent solution, thus the preparation was essentially lipid-free. All molecules were functionally active as shown by substrate binding. PMID- 3042461 TI - Efficient production of native, biologically active human cystatin C by Escherichia coli. AB - A cDNA encoding the mature human cysteine proteinase inhibitor cystatin C was fused to the coding sequence for the Escherichia coli outer membrane protein A signal peptide, and the recombinant gene was expressed in E. coli under the control of the lambda PR promoter, an optimized Shine-Dalgarno sequence and the lambda cI 857 repressor. When induced at 42 degrees C, such cells expressed large amounts of recombinant cystatin C. The recombinant protein was isolated in high yield and characterized. All physicochemical properties investigated, including the positions of disulfide bonds, indicated that the E. coli derived cystatin C was identical to cystatin C isolated from human biological fluids, except that the proline residue in position three was not hydroxylated. The recombinant protein displayed full biological activity against papain, cathepsin B and dipeptidyl peptidase I. PMID- 3042462 TI - Involvement of proteasomes (multicatalytic proteinase) in ATP-dependent proteolysis in rat reticulocyte extracts. AB - The role of proteasomes, particles with latent multicatalytic proteinase, in ATP dependent proteolysis in rat reticulocyte extracts was examined. Removal of proteasomes from the extracts by immunoprecipitation caused almost complete inhibition of ATP-dependent degradation of [3H]methylcasein, without affecting ATP-dependent proteolysis. Peptide fragments of [3H]casein, obtained by cyanogen bromide cleavage, were rapidly degraded in an ATP-independent fashion and this activity was not affected by removal of the proteasomes. These results suggest that proteasomes are involved in ATP-dependent proteolysis in the extracts and that they catalyze the initial cleavage of large proteins. PMID- 3042463 TI - Crystal structure of a complex between thermitase from Thermoactinomyces vulgaris and the leech inhibitor eglin. AB - Thermitase, the thermostable alkaline protease from Thermoactinomyces vulgaris, has been crystallised in a 1:1 complex with eglin, the inhibitor from the medical leech. Two large crystals were grown, with cell dimensions of a = 49.3 A, b = 67.3 A, c = 90.5 A and space group P2(1)2(1)2(1). The crystals are relatively tightly packed with Vm = 2.1 A3/Da. Three-dimensional data to 1.9 A have been recorded from one of these crystals. The orientation and position of the complex in the unit cell have been established using the subtilisin Carlsberg-eglin structure as a model. The structure of the complex is being refined by restrained least-squares. The present crystallographic R factor (= sigma parallel Fo - Fc parallel/sigma/Fo parallel) is 26% at 2.5 A resolution. PMID- 3042464 TI - Analysis of guanine nucleotide bound to ras protein in PC12 cells. AB - The ras gene product (p21) specifically binds GDP or GTP. In analogy with the reaction mechanism of other GTP-binding proteins, only the GTP-bound conformation is believed to be the biologically active one. Previously, we reported that not only oncogenic p21(Val-12) but also proto-oncogenic p21(Gly-12) could induce morphological differentiation in rat pheochromocytoma PC12 cells when microinjected in the complexed form with GTP gamma S [(1987) Mol. Cell. Biol. 7, 4553-4556]. In the present report we transformed PC12 cells with the oncogenic ras gene placed under the metallothionein I promoter. It was found that the transformed cells, when induced with Cd2+, differentiated in the absence of NGF. Then we analyzed the guanine nucleotide bound to p21 in the intact PC12 cells. It was found that conditionally induced p21(Val-12) was mostly present in the GTP bound form, whereas the endogenous p21(Gly-12) was in the GDP-bound form. These results indicate again that p21.GTP induces the morphological differentiation of PC12 cells. PMID- 3042465 TI - Colchicine acts as a progression factor to initiate DNA synthesis in quiescent Balb/c 3T3 cells. AB - In quiescent Balb/c 3T3 cells, competence factors such as 12-O tetradecanoylphorbol-13-acetate (TPA) and platelet-derived growth factor (PDGF) synergize with progression factors such as insulin to initiate DNA synthesis. In this study, we found that colchicine, a microtubule-disrupting agent, acted synergistically with TPA, but not with insulin, to induce the maximal stimulation of DNA synthesis. Colchicine also synergized with PDGF in the presence of epidermal growth factor to elicit nearly the optimal induction of DNA synthesis. Moreover, it acted synergistically with fibroblast growth factor, another competence factor. These results suggest that colchicine acts as a progression factor like insulin in quiescent Balb/c 3T3 cells. PMID- 3042466 TI - rRNA-protein neighbourhood in Escherichia coli 70 S ribosomes and 70 S initiation complex. Probing by bifunctional Pt(II)-containing reagent. AB - The cleavable homobifunctional reagent dichloro[N,N,N',N'-tetrakis(2-aminoethyl) 1,6-hexamethylenediamminedi platinum (II)] dichloride was used for studying rRNA protein cross-links in free 35S-labelled 70 S ribosomes and within initiation complex ribosome.AUGU6.fMet-tRNA(fMet). It was shown that the sets of proteins cross-linked to 16 S and 23 S rRNA in free 70 S ribosomes and in 70 S initiation complex do not differ significantly. The authors are the first to demonstrate most of the 23 S rRNA-protein cross-links and some 16 S rRNA-protein cross-links, in particular those with L7/L12 protein. PMID- 3042467 TI - Different phosphorylated forms of an insulin-sensitive glycosylphosphatidylinositol from rat hepatocytes. AB - Labeling with [3H]galactose was employed to isolate a glycosylphosphatidylinositol from rat hepatocytes which might be involved in the action of insulin. The polar head group of this glycosylphosphatidylinositol was generated by phosphodiesterase hydrolysis with a phosphatidylinositol-specific phospholipase C from Bacillus cereus. By Dowex AG1 x 8 chromatography the polar head group could be separated into three radioactive peaks eluting at 100 mM (peak I), 200 mM (peak II) and 500 mM (peak III) ammonium formate, respectively. Peak III was the most active as an inhibitor of the cAMP-dependent protein kinase. Treatment of peak III with alkaline phosphatase markedly reduced its activity on cAMP-dependent protein kinase. When peaks I, II or III were treated with alkaline phosphatase and analyzed again by Dowex AG1 x 8 chromatography, the radioactivity eluted with the aqueous fraction. The above results indicate that the polar head group of the insulin-sensitive glycosylphosphatidylinositol from rat hepatocytes exists in three different phosphorylated forms and that the biological activity of this molecule depends on its phosphorylation state. PMID- 3042469 TI - A new handheld air impulse tonometer. AB - A new type of non-contact tonometer is described. The instrument is handheld and incorporates a system which automatically generates the measuring pulse of air when the alignment is correct. A brief description of the design and the results of clinical trials are presented. These establish a good correlation with paired Goldmann readings (correlation coefficient, r = 0.88 to 0.95, standard deviation, S.D. 1.56 to 2.66 mm Hg). PMID- 3042468 TI - The muscarinic receptor subtype in mouse pancreatic B-cells. AB - Isolated mouse islets were used to identify the muscarinic receptor subtype present in pancreatic B-cells. We thus compared the inhibitory potencies of atropine (non-specific), of pirenzepine (specific for M1 receptors) and of compound AF-DX 116 (specific for cardiac M2 receptors) on acetylcholine-induced insulin release, 86Rb+ efflux and 45Ca2+ efflux. The three antagonists inhibited all effects of acetylcholine, but EC50 values were markedly different: atropine = 1.5-5 nM, pirenzepine = 0.6-1.7 microM and AF-DX 116 = 1.7-11 microM. The results did not suggest that the various effects of ACh could result from the activation of different subtypes of receptors. It is concluded that muscarinic receptors of pancreatic B-cells belong to an M2 subtype distinct from the cardiac M2 receptors. PMID- 3042470 TI - A comparative evaluation of timolol maleate and pilocarpine in the treatment of chronic open angle glaucoma. AB - Ninety-two eyes with newly-diagnosed chronic open angle glaucoma (COAG) were treated in a randomised prospective trial with either timolol or pilocarpine. Their visual field survival was monitored on a 3-monthly basis over 2 years using both Goldmann and Friedmann perimetry. Concomitant tonometric data was derived by applanation. Fields were assessed and quantified using algorithms designed to give the greatest sensitivity for glaucomatous field loss. Microcomputer programmes specifically designed for this purpose were used in the data collection and subsequent analysis. PMID- 3042471 TI - [Bioactivity and heterogeneity of human growth hormone in urine from acromegalic patients]. AB - Bioactivity of hGH in urine from five acromegalic patients was determined by Nb2 rat lymphoma bioassay (Nb2BA) and IM-9 receptor modulation assay (RMA). One urine sample (case 1) was concentrated by dialysis and lyophilization. Bioactivity by IM-9 RMA showed close correlations with immunoactivity by RIA in serial dilutions (RMA/RIA; 0.66-1.77). The estimates of bioactivity by Nb2BA were higher in all diluted samples except undiluted one than those of immunoactivity. Four samples were concentrated by affinity chromatography, dialysis and lyophilization. The ratio of RMA and RIA estimates was between 0.81-1.24, and that of Nb2BA and RIA estimates was between 0.46-2.62. Heterogeneity of urinary hGH was analyzed by gelfiltration using TSK-G-3000SW column by high performance liquid chromatography (HPLC) and by sensitive sandwich enzyme immunoassay. In addition to the main peak of 22K molecular weight (more than 97%), very small peaks of 40-60K (big hGH) and 150K (big-big hGH) were detected in chromatographic profile. Urinary hGH from acromegalic patients was bioactive in IM-9 RMA and Nb2BA. The predominant form of urinary hGH from acromegalic patients was little hGH, but big and big-big hGH were detected by sensitive sandwich enzyme immunoassay. PMID- 3042472 TI - [Kinetic studies of insulin degrading enzyme in cultured human lymphocytes]. AB - After insulin binds to its receptors, insulin-receptor complexes are internalized by absorptive endocytosis, and then insulin seems to be degraded in the intracellular sites. Although the degradation of insulin has been extensively studied, the sites and enzymes of intracellular degradation have still not been appeared. In order to clarify this problem, following experiments were performed. The effects on insulin degradation of the various inhibitors and anti-IDE serum were investigated in isolated rat hepatocytes and Bri-7 human cultured lymphocytes. N-ethylmaleimide and bacitracin, inhibitors which inhibit the activity of insulin-degrading enzyme (IDE), and anti-IDE serum were decreased insulin degradation in Bri-7 cells which does not contain lysosomal pathway. IDE mainly exists in the cytosol fraction, but also on the surface of various cell types. The kinetic changes of insulin receptors and cell surface IDE was determined in Bri-7 cells after preincubation with or without insulin. The concentration of cell surface IDE was determined by immunoenzymatic labeling method using anti-IDE serum. Bri-7 cells were preincubated with 10(-6) M insulin for 30 min to 18 h. Fifty percent of the receptors were lost at 6 h after the preincubation, and level of the receptors achieved a steady state at 18 h. Although the loss of surface IDE was slightly slower than that of receptors, the curves were essentially parallel to each other. Thus, the loss of cell surface receptors and IDE was directly related to the preincubation time. Furthermore, the recovery of decreased surface receptors and IDE was needed for 36 and 72 h, respectively. The alpha-subunit of insulin receptor (135 K) and IDE (110 K) were assessed by cross-linking of 125I-insulin to the plasma membrane and the cytosol of Bri-7 cells, respectively. Cell surface insulin receptor was decreased, whereas cytosolic IDE was increased in Bri-7 cells incubated with insulin. Thus, it is likely that both cell surface and cytosolic IDE, acting either individually or in concert, constitute a physiological mechanism by which the cellular response to insulin is terminated. These results suggest that IDE may play a major role in insulin degradation in the intact cell. Moreover, one possible mechanism of intracellular insulin degradation is that cell surface IDE may be internalized with the insulin receptor complex and may degrade insulin during the intracellular process. PMID- 3042473 TI - The effect of aging cellular mechanisms on tooth movement. AB - In summary, there appears to be indirect evidence that the dentoalveolar system undergoes a number of changes with age. These changes have the potential of affecting the biologic response of these tissues to the forces of tooth movement. They should not, however, create a barrier toward the successful completion of orthodontic therapy in most cases. PMID- 3042474 TI - Periodontal preparation of the adult patient prior to orthodontics. AB - Evaluation of periodontal status and proposed orthodontic therapy must be made prior to initiating tooth movement. Teeth with a compromised periodontium can be treated. Prior to initiation of orthodontic therapy, it may be necessary to perform periodontal surgery for pocket elimination or reduction, to perform mucogingival surgery, or to correct gross occlusal discrepancies. PMID- 3042475 TI - Oral surgeons' considerations in surgical orthodontic treatment. AB - The orthognathic surgeon seldom has to consider further growth and development of the adult jaws. There are, however, limitations in the adult of certain orthodontic procedures that are effective in the young; rapid palatal expansion, for example. The surgeon and the orthodontist must be aware of other procedures that may be substituted. The adult patient has social, economic, and psychological demands that differ from the young. These may mandate a reversal of the traditional staging of orthodontics first, surgery to follow. Instead, consideration can be given to doing only that orthodontics needed to permit surgery, then surgery to correct the skeletal problems, followed by whatever orthodontics are necessary or desired. Various symptoms of MPD are present in most patients with jaw abnormalities and malrelationships. There must be an awareness that trying to provide relief without correcting the structural problem is treating only symptoms. There also must be realization, especially on the part of the patient, that correcting the jaw deformity does not necessarily mean that symptoms will be gone or, if gone, will not recur. Above all, the orthodontic and surgical team should strive to provide the patient with the maximum function compatible with the appearance the patient desires and do it with the least required amount of surgery and orthodontics. PMID- 3042476 TI - Orthodontic considerations in adult surgical orthodontic cases. AB - The orthodontic considerations in orthognathic cases are based on a detailed understanding of the malocclusion, particularly a differential diagnosis of its facial, skeletal, and dental components, along with a recognition of the limitations of current orthodontic therapy. The goal is to develop a treatment plan that produces the optimal in esthetics, function, and stability while addressing the patient's chief complaint. Attention must be paid to the need to integrate the orthodontic component of treatment with the general dental and surgical phases of the patient's care. PMID- 3042477 TI - Orthodontics and temporomandibular disorders. AB - The presumed relationship between occlusal disharmonies and temporomandibular disorders has been the cornerstone of traditional orthodontic thinking about these disorders. Current research, however, indicates that temporomandibular problems are actually medical orthopedic diseases or dysfunctions that have little to do with occlusal morphology or maxillomandibular relationships. Therefore, orthodontists must discard some of their traditional beliefs and practices, replacing them with modern concepts of musculoskeletal pain and dysfunction, in order to provide appropriate care for patients suffering from temporomandibular disorders. PMID- 3042478 TI - Uses of implants in orthodontics. AB - In recent years, endosseous implants have become practical and reliable adjuncts for dental rehabilitation. That same technology can and will be used in orthodontics to provide anchorage for tooth and bone movement. As anchors for dental movement, osseointegrated implants particularly are useful in patients who are partially edentulous; the implants subsequently can be used as bridge abutments. As anchors for bone movement, implants will be used to modify facial growth. Although implants have not yet been used for this purpose in children, there are no unsolvable technological problems. PMID- 3042479 TI - Treatment of adults with lingual orthodontic appliances. AB - With the advent of lingual orthodontic treatment, an alternative became available to the adult patient who preferred to avoid the unesthetic appearance of conventional orthodontic appliances. The newer brackets and archwires described in this article, in combination with the proven technique developed by the author and others, has made lingual orthodontic treatment a practical reality. The appliance has been shown to be as effective as labial counterparts in correcting all types of malocclusions. New laboratory and indirect bonding techniques have eliminated the need for intricate wire bending and have reduced patient chair time and overall treatment time. Because of the premature introduction of early lingual appliances, many dental practitioners mistakenly believe that lingual treatment is less effective than labial treatment. As more examples of successful treatment are seen, dental practitioners will be more apt to refer patients to orthodontists proficient in this technique. Many graduate orthodontic programs now are teaching this technique to their residents. About 3000 patients currently are starting treatment with lingual appliances each year. This represents only about 1 per cent of adult patients. It is projected that this slowly will climb to about 10 per cent of adult orthodontic treatment over the next 5 years. The increased cost of this treatment, coupled with the resistance on the part of many orthodontists to learn the new technique, seem to be the limiting factors. PMID- 3042480 TI - [Epidermal lectin binding in psoriasis vulgaris. I. Galactosyl glycoconjugates]. PMID- 3042482 TI - Cutaneous malakoplakia. Report of a case. AB - A case of cutaneous malakoplakia in a 44-year-old women under immunosuppressive therapy after renal transplantation for nephrosclerosis is reported. A nodular lesion with inflammatory signs appeared on her right buttock. Histopathology showed a dermal infiltrate with histiocytes containing Michaelis-Gutmann bodies. Electron microscopy showed Michaelis-Gutmann bodies with concentric laminar structure ('target' appearance). Tissue culture revealed Escherichia coli growth. Lesions recurred on surrounding skin and left buttock after excision. Clofazimine treatment resulted in total remission. PMID- 3042481 TI - Familial Becker's nevus. AB - The authors describe 4 patients with Becker's nevus (BN) in two families. Another 4 patients of the german literature are also presented. Together with the first two published cases of BN, familial BN is now known in 10 patients. All three clinical types of BN could be noticed. PMID- 3042483 TI - Oxytocin increases arginine-induced A and B cell secretion in normal man and in diabetic subjects. AB - In previous in Vitro and in Vivo studies oxytocin was shown to stimulate A and B cell secretion. In the present study we show that oxytocin is also able to increase arginine-induced glucagon and insulin secretion in healthy human beings. Similar results were obtained in both insulin-dependent (type-1) and non-insulin dependent (type-2) diabetic subjects. PMID- 3042484 TI - Feasibility of continuous subcutaneous insulin infusion in young diabetic patients. AB - Seven diabetic children (age: 8-12 y.) participated in a study comparing CSII and conventional treatment (CT) under the same monitoring and supervision for a year. The aim of the study was to explore the feasibility, the risks and the results on the glycemic control of CSII. Mean HbAlC fell from 9.3 +/- 2.1% to 7.3 +/- 1.4% (p less than 0.001) after the first month of CSII, remaining stable thereafter and with marginally lower, daily insulin requirements. There was a trend for HbAlc to increase, (HbAlC = 8.6 +/- 1.8%) under conventional treatment. The individual means of premeal capillary blood glucose concentrations were lower under CSII than under CT, but exhibited large variability under both treatments. Symptomatic hypoglycemia was as frequent under CSII as under CT. Ketonuria occurred frequently after the night basal infusion, but was of moderate clinical relevance and mainly due to technical problems. All seven children completed the trial; 4 of them chose to return to pump treatment. No psychologically adverse effects were noted. They all liked the flexibility of lifestyle afforded by CSII, and were generally compliant with the guidelines of this regimen. The pump itself was a valuable educational tool for the children and their families. However, CSII was recognized as individually demanding. This study demonstrates the feasibility of CSII in prepubertal children, and its impact on the glycemic control. The risks of this form of intensified treatment were limited by the strict monitoring and medical supervision provided by a specialized team. PMID- 3042485 TI - Effect of acarbose on glucose and insulin response to sucrose load in reactive hypoglycemia. AB - The aim of the present study was to evaluate the acute effect of acarbose, an alpha-glycosidase inhibitor, on glycemic and insulin response to an oral sucrose load in sixteen patients suffering from idiopathic reactive hypoglycemia. In each subject, two oral sucrose tolerance tests (45 g/m2 body surface) were performed with either placebo or acarbose in a double-blind random order. Compared with placebo, acarbose dramatically flattened the blood glucose curve with values significantly lower from 30 to 90 min and higher from 150 to 240 min. Moreover the magnitude of both glucose peak and nadir, and rate of blood glucose rise and fall were significantly reduced by acarbose as was the hypoglycemic index. Plasma insulin levels from 30 to 120 min, insulin peak and area under the insulin curve were significantly lower after ingestion of acarbose. Thus, in reducing the early rise in blood glucose and insulin after sucrose ingestion, acarbose prevents the occurrence of reactive hypoglycemia; the present study confirms that this drug may be of value in the treatment of reactive hypoglycemia. PMID- 3042486 TI - Purinergic receptors involved in the stimulation of insulin and glucagon secretion. AB - The study of purinergic receptors in the endocrine pancreas is a recent field of investigations. Until recently, ATP has been considered only as an intracellular fuel by most authors. Nevertheless for some years evidence has been obtained that extracellular ATP and/or ADP could act on the B cell membrane to increase insulin secretion. Furthermore it was shown that adenosine could act on the A cell surface to increase glucagon secretion. In both cases the involvement of purinoceptors could be demonstrated using structural analogues which are metabolically stable and not taken up by cells. Also the effects of activators of purinoceptors on B and A cells were suppressed by antagonists. So, it was possible to establish that the purinoceptors involved in the stimulation of insulin and glucagon secretion were different. The B cell is endowed with P2 purinoceptors (for ATP and/or ADP) and the A cell possess P1 purinoceptors (for adenosine). It is of interest to note that B and A cells, which secrete hormones with opposite roles, anabolic and catabolic respectively, have two different types of purinoceptors involved in stimulatory process. The purinergic modulation of B and A cells appears to be of physiological relevance. PMID- 3042488 TI - Remission of insulin dependence induced by continuous subcutaneous insulin infusion (CSII) in type I, recent onset diabetics: role of beta cell recovery and lymphocyte subsets distribution. PMID- 3042487 TI - [Adaptation to sports by insulin-treated diabetics]. AB - Performing muscular exercise regularly is generally recommended to diabetics; indeed, exercise increases muscle insulin sensitivity, helps fighting overweight and, at least partly, tends to correct plasma lipids abnormalities, thus contributing to limit the development of atherosclerosis. Moreover, the practice of sport is beneficial from a psychological point of view, because, thanks to it, diabetic patients can match, even surpass, "the others" and overcome what they often consider as a disability. However, diabetes--especially type 1, insulin dependent, diabetes--deeply modifies the metabolic adaptations to muscular exercise; consequently, exercise must be performed only in good metabolic control conditions, for avoiding a worsening of ketonaemia. In adequately controlled diabetics, muscular exercise can be beneficial by reducing blood glucose levels; it can also lead to hypoglycaemia occurring during or after the exercise bout. In order to reduce the risk of exercise-induced hypoglycaemia, diabetics have to know how to modify three essential parameters of their treatment: (1) increase carbohydrate intake before, during or after exercise; (2) reduce the dose of the insulin acting during exercise, and this in relation to the usual doses and to exercise intensity; (3) under some circumstances, modify the site of insulin injection according to the type of exercise performed. Taking into account these parameters, some general rules can be assessed, which are to be adapted to every particular situation; the use of home blood glucose monitoring before and after exercise is not only useful but sometimes mandatory.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3042489 TI - Insulin and C-peptide responses to oral glucose load in hypothyroid patients. PMID- 3042490 TI - Metabolic and psychological evolution of insulin dependent diabetic patients during a 6 months insulin-pen treatment. AB - The aim of the study is to assess whether an insulin pen-treatment (NovopenR) could be of interest in 10 type I insulin dependent diabetic patients (C-peptide: 0.04 +/- 0.01 pmol/ml, mean +/- SEM), with metabolic and psychological parameters being together taken into account. The daily insulin doses were comparable during the previous treatment with conventional syringes (2-3 daily injections of ActrapidR and MonotardR) and the pen therapy: 0.65 +/- 0.05 vs 0.68 +/- 0.04 U/kg b.w. The metabolic control assessed by HbA1 levels was unchanged before and after 6 months pen treatment: 10.7 +/- 0.7 vs 10.9 +/- 0.6%, respectively. However, fructosamine increased from 2.89 +/- 0.26 to 3.92 +/- 0.20 mmol/l (p less than 0.01) during pen treatment. The psychological objective variables showed no significant changes after pen treatment. In contrast, the staff ratings about the patients attitude toward illness and the spouse evaluation of subjective well being increased from 40.2 +/- 8.4 to 49.5 +/- 7.8 (p = 0.03) and 34.8 +/- 23.4 to 51.1 +/- 21.1 (p = 0.04), respectively. In conclusion, in a limited group of patients, a multiple injection regimen by pen treatment did not lead to an improved metabolic control. However, subjective psychological tests showed that some aspects of well-being tended to improve. PMID- 3042491 TI - Metformin potentiates the antigluconeogenic action of insulin. AB - The interaction of metformin and insulin in the control of hepatic gluconeogenesis was examined in isolated hepatocytes from 48h starved rats using lactate (10(-2) mol/l) with pyruvate (10(-3) mol/l) as substrate. During 1 h incubations in the absence of added insulin, 10(-2) and 10(-3) mol/l metformin reduced gluconeogenesis by 65% and 59% respectively, but lower concentrations of metformin were not effective. Insulin alone (10(-6)-10(-8) mol/l) reduced (37% 16%) gluconeogenesis. The effect of insulin was enhanced (further reductions of 11%-24%) by 5 x 10(-4) mol/l metformin, although this concentration of metformin was not effective in the absence of insulin. At lower insulin concentrations (10( 9) and 10(-10) mol/l) which did not significantly affect gluconeogenesis alone, 5 x 10(-4) mol/l metformin decreased gluconeogenesis (32% and 28% respectively). At 10(-10) mol/l insulin, metformin concentrations of 10(-3)-10(-7) mol/l decreased gluconeogenesis by 60-20%. The results suggest that therapeutic concentrations of metformin may act synergistically with physiological concentrations of insulin to suppress hepatic gluconeogenesis. PMID- 3042494 TI - 'Controlled trial of physical therapy' at Johns Hopkins. PMID- 3042492 TI - Levels of insulin, corticosterone, T3, T4 and insulin sensitivity in fat and lean chickens. AB - The late generations of fat (FL) and lean (LL) chickens were compared. In the fasted state, plasma glucose was lower in FL chickens, whereas insulin, T3, T4 and corticosterone were unchanged. In the fed state, plasma insulin and T4 were increased in FL chickens whereas glucose, T3 and corticosterone were unchanged. During ad libitum refeeding both plasma glucose and insulin, and to a lesser degree T3, were lower and T4 higher in FL chickens whereas corticosterone remained unchanged between lines. The low glucose and insulin levels observed in FL during refeeding, which were not observed in the F4 generation, were also found after force-feeding. In contrast, during oral glucose tolerance test, as in previous generations, a better glucose tolerance, higher plasma insulin and slightly higher free fatty acid levels were found in FL chickens. The hypoglycemic effect of exogenous insulin was very similar (and poor) in both lines in the fed state and was higher in FL than in LL chickens in the fasted state. From the present and previous studies, a change in the glucose-insulin balance and possibly, in T3, could account for the differences in fattening of both lines. PMID- 3042493 TI - Antiprogestins. PMID- 3042495 TI - The effect of blood transfusion on cerebral blood-flow in preterm infants: a Doppler study. AB - Stable infants with anaemia needing a transfusion with adult red blood-cells were studied to elucidate changes in brain blood-flow velocity. Within 24 hours and at five to six days following transfusion a substantial mean flow velocity reduction was observed. Haemodynamic factors contributing to the reduction were an increase in cerebrovascular resistance and an increase in whole blood viscosity, as reflected by a raised pulsatility index. Transfusion with adult red blood-cells causes an elevation in haemoglobin concentration, thereby increasing the total oxygen-carrying capacity of arterial blood; however, this lowers the concentration of fetal haemoglobin which possesses a higher affinity for oxygen. Since cerebral oxygen transport is equal to the product of cerebral blood-flow and arterial oxygen content, this finding suggests the existence of a homeostatic mechanism for cerebral oxygen transport. The actual amount of cerebral oxygen transport was found to increase progressively as the percentage of fetal haemoglobin rose above 30 per cent. At higher fetal haemoglobin levels, appropriate elevations in cerebral blood-flow occurred, causing an increase in the supply of oxygen to the brain. PMID- 3042497 TI - Are complications of pregnancy and birth causes of schizophrenia? PMID- 3042496 TI - Cranial ultrasonography and the prediction of cerebral palsy in infants weighing less than or equal to 1200 grams at birth. AB - The role of serial cranial ultrasonography in the prediction of cerebral palsy was examined in 116 surviving infants with birthweights less than or equal to 1200 g. All underwent serial real-time sonographic examinations of the brain on days one, five and 21, then monthly, until term corrected age. Intraventricular hemorrhage (IVH) was diagnosed in 48 infants, and three had periventricular leukomalacia. Of the 116 infants, 31 had ultrasound abnormalities at term. At 12 to 18 months corrected age 12 infants had cerebral palsy and 38 were classified as suspect; the other 66 were normal. There was a clear association between risk group, based on sonographic findings at term, and outcome. Infants with IVH whose cranial ultrasounds failed to become normal by term corrected age were at higher risk for cerebral palsy than those with normal examinations at term, regardless of the severity of IVH. Thus an abnormal ultrasound at term corrected age was highly predictive of cerebral palsy, especially among survivors of IVH. It remained the best predictor of cerebral palsy, even when other perinatal and neonatal variables were considered. In contrast, duration of mechanical ventilation, rather than sonographic findings, was the best predictor of suspect neuromotor status. PMID- 3042498 TI - Selective dorsal rhizotomy for spastic cerebral palsy. PMID- 3042499 TI - The classification of children with reading difficulties. PMID- 3042500 TI - Subsequent management of children with febrile convulsions. PMID- 3042501 TI - Scintigraphic study of duodenogastric reflux. Value of a computerized image subtraction method. AB - Duodenogastric reflux (DGR) could be implicated in several esophageal and gastric diseases. Establishing its pathophysiological role however is difficult because of the problems in the demonstration and quantification of DGR episodes. The aim of this study was to improve a scintigraphic method of detection and quantification of DGR episodes during the postprandial period in man. The study was carried out in 14 control subjects (7 males and 7 females, median age = 25 years, range: 22-35 years). As scintigraphic recording was continuous during 150 min, all DGR episodes were revealed. In order to improve visual detection of DGR episodes, images were treated by a computerized image subtraction method. The visual detection limit of DGR episodes determined by comparison to test images was 0.6 p. 100 of the dose injected intravenously or 17 microCi. A DGR episode was demonstrated in one of the 14 control subjects. The quantity of refluxed liquid was estimated, in this case, at 30 microCi, and the duration of the reflux greater than 2 min. Continuous scintigraphic recording in association with a computer based technique of image subtraction seems to improve scintigraphic performance in the study of DGR episodes under pathological conditions. PMID- 3042502 TI - [Resection of the rectum with colo-anal anastomosis in the treatment of the cancer of the rectum]. PMID- 3042503 TI - [Peptides of the bombesin family: from their physiology to potential therapeutic applications]. PMID- 3042504 TI - [Study of insulin resistance in cirrhosis by the hyperinsulin euglycemia clamp]. AB - To explore the insulin resistance state in liver cirrhosis and to assess the respective role of insulin sensitivity and cellular metabolism alterations, an euglycemic glucose clamp was performed in 12 cirrhotic patients and 6 healthy volunteers with 3 successive hyperinsulinemic periods of 90 mn (infusions of 7, 20 and Iu/h). An artificial beta-cell model allowed to quantify the amount of glucose needed to keep glycemia at 4.70 mmol/l. In 7 cirrhotic patients, insulin secretion was tested by an intravenous glucose tolerance test. The dose-response curves showed a significant (p less than 0.01) decrease of the theoretical maximal metabolic clearance rate of glucose (capacity). The insulin concentration corresponding to the half-maximal response (ED50) did not differ between cirrhotic and control subjects because of a very important dispersion of individual values in cirrhotics. The ED50 value was lower than the values of control subjects in five cirrhotic patients and normal or enhanced in seven other patients; the former showed the most reduced values for capacity, and the latter a more marked hyperinsulinemia during the intravenous glucose tolerance test. The two subgroups of patients did not differ for clinical or biological parameters of cirrhosis, or for glucose tolerance. These results 1) show a constant and marked impairment of glucose metabolism capacity in liver cirrhosis, 2) and suggest that the insulin resistance inconstantly observed in this state is a consequence of chronic hyperinsulinemia. PMID- 3042505 TI - [Perendoscopic biliary manometry]. PMID- 3042506 TI - Eighty years ago: the beginnings of population genetics. PMID- 3042507 TI - A fourth Escherichia coli gene system with the potential to evolve beta-glucoside utilization. AB - Escherichia coli K12 is being used to study the potential for adaptive evolution that is present in the genome of a single organism. Wild-type E. coli K12 do not utilize any of the beta-glucoside sugars arbutin, salicin or cellobiose. It has been shown that mutations at three cryptic loci allow utilization of these sugars. Mutations in the bgl operon allow inducible growth on arbutin and salicin while cel mutations allow constitutive utilization of cellobiose as well as arbutin and salicin. Mutations in a third cryptic locus, arbT, allow the transport of arbutin. A salicin+ arbutin+ cellobiose+ mutant has been isolated from a strain which is deleted for the both the bgl and cel operons. Because the mutant utilized salicin and cellobiose as well as arbutin, it is unlikely it is the result of a mutation in arbT. A second step mutant exhibited enhanced growth on salicin and a third step mutant showed better growth on cellobiose. A fourfold level of induction in response to arbutin and a twofold level of induction in response to salicin was observed when these mutants were assayed on the artificial substrate p-nitrophenyl-beta-D-glucoside. Although growth on cellobiose minimal medium can be detected after prolonged periods of time, these strains are severely inhibited by cellobiose in liquid medium. This system has been cloned and does not hybridize to either bgl or cel specific probes. We have designated this gene system the sac locus. The sac locus is a fourth set of genes with the potential for evolving to provide beta-glucoside utilization. PMID- 3042508 TI - Isolation and characterization of mutants which show an oversecretion phenotype in Saccharomyces cerevisiae. AB - We have isolated mutants responsible for an oversecretion phenotype in Saccharomyces cerevisiae, using a promoter of SUC2 and the gene coding for alpha amylase from mouse as a marker of secretion. These mutations defined two complementation groups, designated as ose1 (over secretion) and rgr1 (resistant to glucose repression). The ose1 mutant produced an oversecretion of amylase by 12- to 15-fold under derepressing conditions; however, the amylase mRNA was present at nearly the same amount as it was in the parent cells. No expression of the amylase gene was detected under repressing conditions. The rgr1 mutant oversecreted amylase by 11- to 13-fold under repressing conditions by 15- to 18 fold under derepressing conditions. The rgr1 mutant showed pleiotropic effects on the following cellular functions: (1) resistance to glucose repression, (2) temperature-sensitive lethality, (3) sporulation deficieny in homozygous diploid cells, and (4) abnormal cell morphology. The rgr1 mutation was not allelic with ssn6 and cyc9, and failed to suppress snf1. PMID- 3042509 TI - Coincident recombination during mitosis in saccharomyces: distance-dependent and independent components. AB - In mitosis, coincident recombination events between widely separated markers occur more frequently than expected for two independent acts. Several different mechanisms have been proposed to account for this phenomenon. It has been argued that coincident recombination could be due to either an extensive region of heteroduplex DNA or some other distance-dependent mechanism. Alternately, it has been suggested that at least some is due to subpopulations of cells which undergo recombination at very high frequencies. The purpose of these experiments is to evaluate the possible contribution of distance-dependent and distance-independent components. By comparing the coincident recombination frequencies for markers on the same homolog as well as pairs of unlinked sites, we show that there is a strong distance-dependent component for at least 8.8-35-kbp, depending on the type of recombination event (conversion or intrachromosomal exchange). For larger distances separating sites, a distance-independent mechanism(s) results in higher than expected frequencies. PMID- 3042511 TI - 'Catastrophic' coverage: good, but we can do better. PMID- 3042510 TI - High-level expression of the cloned ada gene of Escherichia coli by deletion of its regulatory sequence. AB - The Ada protein, a methyltransferase for repair of several alkyl adducts in DNA, was expressed in its native form at a high level in Escherichia coli from a pUC9 recombinant plasmid carrying ada gene from which the sequence controlling the Ada induction was deleted. The regulatory sequence appears to act as a terminator of transcription initiated from the lac promoter of the vector. However, deletion of the regulatory sequence resulted in elimination of ada induction by alkylating agents, providing confirmation of its role in activation of ada expression. PMID- 3042512 TI - Home health care: what's available and what Medicare pays for. PMID- 3042513 TI - Practical management of catheter-associated UTIs. AB - In the long-term catheterized elderly, damage to the epithelial wall is primarily responsible for bladder infections, due to the presence of the catheter as a foreign body and its frequent manipulations. The presence of bacteriuria, a classic sign of infection in the non-catheterized patient, is of little diagnostic importance in the long-term catheterized patient, since it is both permanent and inevitable. Routine preventive measures--eg, frequent catheter changes, prophylactic antibiotics--are of no value. Other measures are worth considering and, if instituted, will reduce the frequency of severe complications from long-term catheterization and improve quality of life. PMID- 3042514 TI - Practical overview of sexual function and advancing age. AB - Sexuality remains a vital aspect of human life well into advanced age. Previous sexual history, opportunity, and overall health status determine whether the sexual activity option is exercised. Normal aging is associated with some physiological changes in genital function. If elderly patients seek help for sexual problems, therapeutic interventions focus on elimination or alleviation of physical and pharmacological interferences, as well as on optimal adaptation to physiologically altered responses. PMID- 3042516 TI - [Friedrich Nietzsche--a forerunner of psychoanalysis?]. PMID- 3042515 TI - Use oral hypoglycemics with caution...but keep caution in context. PMID- 3042517 TI - [The discussion on euthanasia in France and Switzerland before World War II]. PMID- 3042519 TI - [Medical management, health conditions and human experimentations at the Ravensbruck female concentration camp]. PMID- 3042518 TI - [The health poem of Burkhard von Horneck (1522)]. PMID- 3042521 TI - The development of sulfonamides (1932-1938) as a focal point in the history of chemotherapy. PMID- 3042520 TI - Another minority's role in medicine: the Armenians. PMID- 3042522 TI - [Julien Barry and the "neurosecretory synapses" (1954-1973)]. PMID- 3042523 TI - Monkey trabecular meshwork cells in culture: growth, morphologic, and biochemical characteristics. AB - We established tissue cultures of trabecular meshwork cells from cynomolgus monkey eyes. The cultures were initiated within 4 h of enucleation on Falcon Primaria flasks. Using medium containing 10% fetal bovine serum and 5% calf serum, trabecular meshwork cells could be grown for up to eight passages without additional growth factors. The growth pattern and cell morphology were distinct from those seen in fibroblastic or endothelial cultures derived from neighboring tissues. Ultrastructurally, our cells showed the characteristics of trabecular meshwork cells, exhibiting prominent basement membranes, intercellular junctions, pinocytotic vesicles, microvillous projections, and branched cell extensions. These cells were grown mostly as monolayers. However, they also appeared to form multi-layered arrays in densely confluent areas when plated at a high density. The extracellular matrix material was surrounded by cells and cell processes, simulating in vivo trabecular beam formation. Radiolabeling experiments demonstrated that our trabecular meshwork cells had the capacity to produce collagen. These results indicated that our cultured cells retain many in vivo characteristics and may be used for various biologic studies of trabecular meshwork. PMID- 3042524 TI - Reproducibility of computerized pallor measurements obtained with the Rodenstock Disk Analyzer. AB - Computerized pallor measurements of the optic nervehead were performed by global analysis with the Rodenstock Disk Analyzer. Total intervideographic and intravideographic variability was calculated. The coefficients of variation ranged from 8.3% to 20.08% for intervideographic variability of different pallor histogram values. The intravideographic variability for pallor histogram values ranged from 0.82% to 2.94%. PMID- 3042525 TI - Increase of corneal graft survival by use of topically immunosuppressive agents in rabbits. AB - Corneal graft survival in rabbits was significantly prolonged by topical treatment of the recipient eye with cyclophosphamide (0.1% in sterile isotonic NaCl) and methotrexate (0.1% in arachis oil) applied four times daily for 4 weeks. A control group of animals was treated with dexamethasone in the same way. The mean graft survival time was 52 +/- 18.5 days in the group treated with dexamethasone and 80 +/- 18.2 and 73 +/- 23.5, respectively, in the groups treated with cyclophosphamide and methotrexate. Cyclophosphamide and methotrexate applied topically produced no systemic side effects, and only one mild and transient local side effect in the form of hyperemia of the bulbar and palpebral conjunctiva. These two immunosuppressive drugs were more effective than the corticosteroid currently in clinical use, i.e., dexamethasone. PMID- 3042526 TI - [Use of liquid chromatography in hygienic studies]. PMID- 3042527 TI - [Hygienic aspects of the study of carcinogenic N-nitroso compounds]. PMID- 3042528 TI - [Heightened individual sensitivity to noise and methods of its detection]. PMID- 3042529 TI - [Current aspects of the water supply activity of WHO]. PMID- 3042530 TI - [Complex study of the state of health of the population in regions using pesticides extensively]. PMID- 3042531 TI - Opioid-serotoninergic dysregulation in the pathophysiology of Tourette's syndrome. AB - In the following communication the major neurotransmitter systems postulated to be involved in the pathophysiology of Tourette's syndrome are discussed. Malfunctions of the opioid and serotoninergic systems are discussed regarding their role in the pathophysiology of the neuropsychological, behavioural and other manifestations of the disease. PMID- 3042532 TI - [Bern Breast Symposium. Festschrift for Max Berger. 22 September 1987, Bern. Proceedings]. PMID- 3042533 TI - [Problems in the occurrence of radiologically detected, nonpalpable breast changes]. PMID- 3042534 TI - [Surgical therapy of breast cancer]. PMID- 3042536 TI - [Adjuvant treatment of breast cancer]. PMID- 3042535 TI - [Precancerous conditions of the breast]. PMID- 3042537 TI - [Questions on hormone and chemotherapy in metastatic breast cancer]. PMID- 3042538 TI - [Radio-oncologic therapy of breast cancer]. PMID- 3042539 TI - [Histopathology of the chorionic villi in the first trimester of pregnancy]. PMID- 3042540 TI - [Diagnosis on chorionic villi tissue in the 1st trimester--rapid karyotyping in the 2d and 3d trimesters]. PMID- 3042541 TI - [Biochemical diagnosis of fetal cells]. PMID- 3042542 TI - [Assessing the fetal movement pattern in disorders of the nervous system]. PMID- 3042543 TI - [Sampling of fetal blood]. PMID- 3042544 TI - [Prenatal treatment of fetal heart diseases]. PMID- 3042545 TI - [Intrauterine therapy of the 21-hydroxylase defect (congenital adrenogenital syndrome)]. PMID- 3042546 TI - [Neurosurgical therapy in congenital disorders of the central nervous system]. PMID- 3042547 TI - [Surgical therapy of prenatally detected abdominal abnormalities]. PMID- 3042548 TI - [Congenital thoracic abnormalities and abnormalities of the intrathoracic organs]. PMID- 3042549 TI - [Ethical considerations in prenatal diagnosis]. PMID- 3042550 TI - [Legal aspects of the diagnosis and therapy of intrauterine fetal disorders]. PMID- 3042551 TI - [Pain therapy in gynecologic malignancies]. PMID- 3042552 TI - [Genetic counseling and indications for prenatal diagnosis]. PMID- 3042553 TI - [Rate of spontaneous abortion in early pregnancy]. PMID- 3042554 TI - [Cytogenetic aspects of chorionic villi biopsy]. PMID- 3042555 TI - Placental protein 21. Localization in human placenta and concentrations in the body fluids of men and nonpregnant and pregnant women. AB - The immunohistochemical localization of placental protein 21 (PP21) was marked in the syncytial brush border and basal membrane during the 1st and 2nd trimesters of pregnancy and also in the chorionic epithelial brush border and basement membrane at term. A weaker stain was found in the cell membranes of amniotic epithelial and chorionic trophoblast cells. Neither heparin nor changes in temperature significantly influenced PP21 concentration. Relatively high serum PP21 concentrations were measured during the follicular and luteal phases in healthy nonpregnant women and in healthy men whose seminal plasma also showed a high PP21 concentration. Serum PP21 levels in normal pregnancy rose from a median of 29.1 ng/ml at 6-7 weeks of gestation to 82.0 ng/ml at 36-37 weeks of gestation. Although maternal urine showed low PP21 levels during pregnancy, amniotic fluid PP21 levels were higher at 7-21 weeks of gestation than at term. Cord blood sera showed almost the same PP21 concentration as maternal sera, but retroplacental blood showed much higher levels. Maternal serum PP21 levels in hydatidiform mole patients did not differ from the normal pregnancy range, although their molar vesicular fluids contained higher PP21 concentrations. These results suggest an extraplacental source for PP21. PMID- 3042556 TI - Succenturiate placenta diagnosed by ultrasound. AB - The succenturiate placenta is a morphological abnormality, the antenatal recognition of which is important as vessels connecting the main placenta with the succenturiate placenta may rupture during labor and fetal death may ensue. In addition, retention of the placental material may lead to postpartum hemorrhage. We treated 5 patients with a succenturiate placenta and the antenatal ultrasonograms and related discussions are presented herein. PMID- 3042557 TI - [Clinical significance of zinc]. PMID- 3042558 TI - [Depression and suicide in physical diseases]. PMID- 3042559 TI - [Oral premedication before ENT interventions with local anesthesia. Effect profile of midazolam, flunitrazepam and morphine sulfate in comparison with placebo]. PMID- 3042560 TI - [Acute treatment of duodenal ulcer. Results of an open multicenter study with famotidine nocte]. PMID- 3042562 TI - [Vasodilator therapy in chronic heart failure]. PMID- 3042561 TI - [Psychotropic drugs in pregnancy and breast feeding. A review of the literature]. PMID- 3042563 TI - [Beta 2 microglobulin. 1: Physiologic significance, behavior in kidney diseases and chronic hemodialysis]. PMID- 3042564 TI - [Beta 2 microglobulin. 2: Dialysis-associated amyloidosis]. PMID- 3042565 TI - [Disorders of sexual function caused by drugs]. PMID- 3042566 TI - [Changes in the venous system of the leg in pregnancy. Diagnosis and use of noninvasive methods]. PMID- 3042567 TI - [Spontaneous remissions in psychoneurotic diseases. Significance for the theory and practice of psychotherapy]. PMID- 3042568 TI - Blood binding and tissue uptake of drugs. Recent advances and perspectives. AB - The free drug hypothesis, which states that only the unbound moiety of drug in blood is available for tissue diffusion, is discussed according to recent investigations. In some experimental conditions, it must be assumed that part of the protein-bound drug in plasma is extracted during a single passage through the organ studied. The mechanisms underlying these observations are not unequivocal and remain hypothetical. In the liver, high-affinity binding sites for serum albumin have been demonstrated, and they would explain the high extraction by liver of endogenous and exogenous compounds. However, these experiments measure the unidirectional transfer of a drug from the vascular to the extravascular space in non-steady-state conditions. Hence, in steady-state conditions, the free drug hypothesis cannot be ruled out because it is supported by numerous pharmacokinetic studies. PMID- 3042569 TI - Drug pharmacokinetics in the obese. AB - In the obese, modifications in body constitution (higher percentage of fat and lower percentage of lean tissue and water) can affect drug distribution in the tissues. For slightly liposoluble molecules (e.g., digoxin, antipyrine), the equilibrium distribution volume (V), total and per kilogram weight, is significantly less than that of control subjects. With lipophilic drugs (e.g., barbiturates, benzodiazepines), this parameter is significantly increased, explaining the prolongation of the plasma elimination half-life. For drugs that are almost equally soluble in water and oil (methyl xanthines, aminoglycosides), the V is slightly increased in the obese. The other main factors involved in drug diffusion in the tissues are binding to plasma and tissue proteins, and regional blood flow. In the obese the binding of drugs to albumin does not seem to be altered. A marked increase in plasma alpha-glycoprotein acid and in propranolol binding has been reported in some studies; this has not been corroborated by other authors. Although the cardiac output and total blood volume are increased in the obese, the blood flow per gram of fat is less than in nonobese subjects. This could limit diffusion in the tissues of some lipophilic drugs. Studies on hepatic clearance of drugs are not available in the obese, but hepatic histological alterations have been described. In most publications concerning drugs with biotransformation as the principal elimination route, the total plasma clearance is not reduced. Up to the present, there are no reports of any impairment involving renal elimination of drugs in the obese. Dose-adjustment of hydrophilic drugs is assessed according to the ideal weight of the individual obese subject; with lipophilic drugs the loading dose can be fixed according to the total weight; calculation of the maintenance dose depends on possible changes in the total clearance. PMID- 3042570 TI - [Pathophysiologic principles of ventricular late potentials--mechanisms]. AB - The majority of sustained ventricular tachycardias are caused by a reentry mechanism. Conditions prerequisite to a reentry circuit are electrophysiological changes, namely conduction delay and block which are incurred by virtue of changes in the myocardial tissue due to scarring, ischemia or fatty infiltration and result in an abnormal action potential, increased intracellular resistance and reduction in intercellular coupling (intercalated discs). Additionally, the principle of anisotropy, that is, the varying velocity of conduction parallel and perpendicular to the fiber axis (increased axial resistance) can also be responsible for circling movement. A further factor predisposing to ventricular tachyarrhythmias is the heterogeneity of repolarization. Delayed and inhomogeneous spread of conduction in morphologically altered tissue can be detected invasively as delayed and fragmented electrograms during sinus rhythm and induced ventricular tachycardia and noninvasively as high-frequency signals which begin in the terminal portion of the QRS complex and can extend far into the ST segment. In a study on our service, electrograms from post-infarct patients with sustained ventricular tachycardia were compared with those without ventricular arrhythmias. In patients with ventricular tachycardia there was markedly delayed activation (greater than 100 ms after begin of the QRS complex, mean 137 ms) over the infarct area. In patients with ventricular arrhythmias, three electrogram types could be differentiated: 1. a delayed singular deflection at the end of the QRS complex (late potential); 2. a diminished amplitude of the electrogram as well as fragmentation into many single deflections for a duration of more than 100 ms; 3. two deflections clearly distinguishable from each other. All three electrogram types represent a different extent of delayed conduction. With one singular late potential there is homogeneous conduction delay throughout the entire region of tissue under the electrode; with fragmentation there is desynchronization of the spread of conduction; with double potentials there are structures of two different velocities of conduction next to or above each other. In almost all patients with ventricular tachycardias fragmented electrograms could be registered in a circumscribed region, in more than one-half there were double potentials. The areas from which abnormal electrograms could be registered were significantly larger in patients with ventricular tachycardias than in those without tachycardia.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3042571 TI - [Methods for the detection of ventricular late potentials. High amplification ECG, signal averaging technic, frequency analysis and intracardiac mapping]. AB - Circumscribed areas of injured myocardium which lead to late ventricular depolarization represent the pathologic-anatomic substrate for reentry mechanisms potentially capable of propagating ventricular tachycardia at the ventricular level. If the myocardial area from which delayed ventricular depolarization and, consequently, late potentials eminate, exceeds a critical minimal size, documentation of such signals can not only be achieved with direct endocardial mapping or catheter mapping but also by means of special high-resolution ECG techniques from the body surface. Since high amplification of the conventional ECG results in registration of noise signals in amplitude of up to 50 microV, late potentials with their amplitudes at the body surface ranging from 5 to a maximum of 20 microV, can only be discriminated after substantial enhancement of the signal-to-noise ratio. The noise arises from no less than three sources: physiologic noise, for example, from muscle activity; electronic noise from amplifiers and background noise of 50 or 60 Hz, respectively. To improve the signal-to-noise ratio, currently three methods are employed: sequential or temporal signal averaging, spatial signal averaging and fast Fourier transformation analysis of the frequency spectrum of the highly-amplified ECG. Temporal signal averaging has the purpose of smoothing randomly-occurring background noise and, at a specified point in time of the ECG cycle, to sum the signal incurred. The effectivity of this technique, however, is subject to certain conditions: the signal to be registered and the background noise must be independent from each other, the noise must be stationary and show normal random distribution, the signal of interest must be periodic and/or coupled with a fixed interval to a point in the ECG cycle which can be used as a trigger. The quality of the averaged signal is dependent on trigger stability. There are three approaches to trigger processing: voltage threshold determination, slope detection and the pattern matching technique, the accuracy, reliability and time consumption of which increases in the order listed. A trigger stability of +/- 0.5 ms is necessary to detect ventricular late potentials with sufficient sensitivity and without meaningful deformation or attenuation of their form and temporal extent. Intercurrently, a number of commercially-acquirable signal averaging computers have been made available which differ with respect to registration and analysis.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3042572 TI - [Ventricular late potentials in acute myocardial infarct]. AB - Ventricular late potentials are regarded as an expression of delayed impulse conduction in an area of myocardial ischemia and, accordingly, indicative of a preformed reentry circuit. Late potentials can be detected in chronic, stable coronary artery disease and their presence correlates closely with impairment of ventricular function and with the probability of future occurrence of tachyarrhythmic events or sudden cardiac death. While repetitive ventricular arrhythmias in the chronic stage of coronary artery disease result almost invariably from circling intraventricular wavefronts, tachyarrhythmias associated with acute myocardial infarction appear attributable to differing pathomechanisms. According to experimental studies, in acute myocardial infarction, three phases of arrhythmogenesis can be differentiated: phase 1 encompasses the first hours after vessel occlusion which generally corresponds with the prehospital phase. Due to the difference in potential of up to 25 mV between ischemic and nonischemic cardiac muscle areas, an injury current is called into existence which leads to depolarization of normal cardiac muscle tissue. The ectopic impulses so precipitated, the conduction of which is supported by the functional inhomogeneity of the infarcted region, are capable of initiating reentry tachycardia. During phase 2, a few hours to days after the ischemic event, only the subendocardial Purkinje fibers in the infarcted region exhibit focal arrhythmogenicity. In contrast to the working myocardial cells, the latter survive due to their immediate proximity to the cardiac chamber and show, ischemia-induced, a propensity to high-frequency impulse formation in terms of abnormal automaticity. Similar to the experimental findings, the cause of the frequently-observed ventricular arrhythmias in the early hospital phase appears predominantly attributable to a focal arrhythmia mechanism. During phase 3, several days to weeks after the acute myocardial ischemic event, reentry mechanisms again are in the foreground in which the electrophysiologic changes in the Purkinje fibers, in terms of increasing desynchronization, together with conduction barriers arising through the infarct scar, pave the way for reentry phenomenon. After abrupt restoration of patency of a previously occluded vessel the very frequent "reperfusion arrhythmias" are also attributable primarily to reentry mechanisms due to inhomogeneous improvement of the conduction properties in the region of the reperfused myocardium. Ventricular late potentials can be registered both invasively by means of epi- or endocardial leads as well as noninvasively from the body surface.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3042573 TI - Prognostic significance of ventricular late potentials in the postmyocardial infarction period. AB - Ventricular late potentials in patients after myocardial infarction can be assumed to herald an increased risk of future sudden cardiac death or symptomatic sustained ventricular tachycardia. This holds particularly true for patients studied early after recent myocardial infarction whereas patients assessed later in the subsequent course have a substantially lesser incidence of arrhythmic events, probably due to intercurrent death of those at high risk. Of prognostic importance appears not only the presence but also the duration of late potentials. A meaningful role is also assumed by the extent of left ventricular functional impairment (EF less than 40%). However, in consideration of the complex mechanisms that can lead to sudden cardiac death, no single method predicts with high sensitivity the occurrence of a ventricular tachyarrhythmic event. Sudden cardiac death can be incurred on the basis of chronic electrophysiological abnormalities as a consequence of regional slow conduction in the border zone of a previous myocardial infarction precipitated by trigger factors such as spontaneous ventricular arrhythmias. Sudden cardiac death or symptomatic sustained ventricular tachycardia can also occur due to sudden and transient changes in the electrophysiological properties of the myocardium due to ischemia. Whether the combination of late potentials with clinical parameters such as ventricular arrhythmias detected in the ambulatory ECG and those induced with programmed electrical stimulation will lead to more accurate identification of patients at risk prerequisites further elucidation. Currently available literature indicates that in patients with late potentials, ventricular tachycardias can be induced more frequently by programmed electrical stimulation and that the combination of both phenomena confers a particularly high risk. PMID- 3042574 TI - Effects of non-pharmacological interventions on ventricular late potentials. AB - The purpose of the paper is to review the presently available information of the effects of non-pharmacological interventions for the control of drug-refractory ventricular tachyarrhythmias on non-invasively recorded ventricular late potentials. During recent years, non-pharmacological interventions have evolved as an alternative form of treatment to control drug-refractory ventricular tachyarrhythmias. The effects of these non-pharmacological measures on ventricular late potentials are poorly understood. Successful surgical control of ventricular tachycardia often normalizes the signal averaged ECG and may eliminate delayed potentials. Thus, this non-invasive test may be useful in assessing surgical efficacy in subgroups of patients with ventricular tachycardia. However, the clinical value of late potentials in assessing surgical efficacy in the individual case may be limited as the sensitivity and specificity of the loss of late potentials after antitachycardia surgery are low. In addition, the effects of transvenous catheter ablation on ventricular late potentials will be reviewed. The available information suggests that this intervention has little effects on the presence or absence of late potentials. Thus, non-invasive recording of late potentials seems not to be helpful in predicting the acute and long-term efficacy of catheter ablation. In conclusion, changing of the parameters in the signal-averaged QRS complex after antitachycardia surgery may be useful in predicting the efficacy of surgical interventions for drug-refractory ventricular tachycardias in subgroups of patients. However, the sensitivity and predictive accuracy of this test are low, thus limiting its clinical usefulness. PMID- 3042575 TI - The significance of high-density lipoproteins (HDL) in the clearance of intravenously administered bacterial lipopolysaccharides (LPS) in mice. AB - The direct immunofluorescence technique was used to study the presence of high density lipoproteins (HDL) in the liver, spleen and kidney of mice before and after intravenous administration of purified lipopolysaccharides (LPS) from Escherichia coli O 75, Salmonella abortus equi and Salmonella minnesota R 595, as well as free lipid A. Untreated mice had small granules of HDL in the hepatocellular cytoplasm, which appeared more pronounced after oral administration of fat. After intravenous administration of LPS, hepatocellular HDL decreased continuously and was no longer visible 1 hour after injection of LPS or lipid A. Five to ten minutes after administration, smooth-form LPS were located in sinusoidal cells, and rough form LPS and lipid A were found in both parenchymal and nonparenchymal liver cells. All toxins were demonstrated in both cell populations 1-4 hours after injection. The spleen was free of HDL, while there was a strong uptake of LPS into the cells of the reticulo-endothelial system. The kidney of experimental mice had small HDL granules in the epithelial cells of the proximal tubules, with a tendency to move to the lumen 1 hour after LPS injection, while there was a transient granular deposition of LPS in the glomeruli. The results suggest that the early uptake of circulating LPS by cells of the reticulo-histiocytic system in liver and spleen, as well as by hepatocytes, is not mediated by HDL. However, HDL located in hepatocytes or present in the circulation of experimental mice seem to be eliminated through the bile and the urine, induced by LPS. PMID- 3042576 TI - Cisapride versus ranitidine in the treatment of reflux esophagitis. AB - The healing effect of the prokinetic drug cisapride (10 mg q.i.d.) on esophageal lesions, and its therapeutic control of gastroesophageal reflux symptoms were compared with the effects of the H2-antagonist ranitidine (150 mg b.i.d. + placebo b.i.d.) in a double-blind trial. In each group, 28 patients with Savary Miller Grade I or II esophagitis were treated for 6 or 12 weeks. At the end of treatment, follow-up endoscopy showed that mucosal lesions were absent in 89% of the cisapride patients and in 79% of the ranitidine patients. In addition, 86% and 82% of the patients in the cisapride and the ranitidine group, respectively, had no, or only mild, reflux symptoms. Minor side effects were experienced in both groups. From these data, cisapride appears to be as effective as ranitidine in controlling reflux symptoms and in promoting the healing of mucosal lesions in milder forms of reflux esophagitis. PMID- 3042577 TI - High serum levels of DUPAN2 antigen and CA19-9 in pancreatic cancer: correlation with immunocytochemical localization of antigens in cancer cells. AB - We determined the concentration of serum DUPAN2 and CA19-9 in 43 pancreatic cancer patients by enzyme immunoassay, and compared the staining patterns of the antigens in the tissues of pancreatic cancer in order to clarify the mechanism of the elevation of serum DUPAN2 and CA19-9 levels in the patients. In 26 patients (60%), the serum DUPAN2 concentration was higher than 300 U/ml. This positive rate was not so high as that of CA19-9. However, seven out of the twelve CA19-9 negative patients were DUPAN2-positive. Using a combined assay of serum DUPAN2 and CA19-9, the diagnostic sensitivity for pancreatic cancer was 88% (38/43). Immunocytochemically, both DUPAN2 and CA19-9 were restricted to the apical surface and/or supranuclear cytoplasm in the normal pancreatic duct epithelia. In cancerous glands, however, the two were found over the entire surface and cytoplasm of the cancer cells--losing the polar distribution pattern of the antigen--and in the surrounding stroma adjacent to the cancer cells. DUPAN2 was detected in 47 (89%) out of 53 adenocarcinomas of the pancreas, and CA19-9 in 44 cases (83%). Cases with high serum antigen levels tended to display high proportions of stromal staining in the cancer tissues. These findings suggest that shedding of the antigen into the stroma adjacent to the malignant glands is one of the major mechanisms in the elevation of high serum DUPAN2 and CA19-9 levels. PMID- 3042578 TI - Insulin modulates early-phase noradrenaline response to glucose ingestion in humans. AB - To clarify the relationship between the early-phase insulin response and the early-phase noradrenaline (NA) response to glucose ingestion in humans, serum NA, adrenaline, immunoreactive insulin (IRI), C-peptide immunoreactivity, potassium, nonesterified fatty acid and plasma glucose levels were measured in 8 non diabetics and 10 diabetics without autonomic disturbance after oral 75 g glucose load. Following results were obtained: 1) In non-diabetics, the maximal NA response was observed at 30 min after glucose ingestion, but in diabetics, mean serum NA levels remained unchanged. The effect of glucose ingestion on the NA response was significantly different between non-diabetics and diabetics by the repeated measurements analysis of variance (F ratio = 5.72, P less than 0.05). 2) In total group (n = 18), at early-phase after glucose ingestion (at 30 min), positive correlation was found between dIRI level and dNA level (r = 0.52, P less than 0.05), between dIRI level and %dNA level (r = 0.56, P less than 0.05), between dIRI/dglucose ratio (insulinogenic index) and dNA level (r = 0.70, P less than 0.01). 3) In four diabetics, NA responses to glucose ingestion were studied again after mild energy restriction for 2 wk. In three of them, both early-phase IRI response and early-phase NA response to glucose ingestion improved after diet therapy, but in the remainder, early-phase NA response to glucose ingestion remained unchanged in accordance with sustained impaired early-phase insulin response to glucose ingestion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3042580 TI - Increased renin release by exogenous prostaglandin E1 in liver cirrhosis with and without ascites. AB - To evaluate the sensitivity of the renin-angiotensin-aldosterone system in patients with liver cirrhosis, prostaglandin E1 was intravenously administered at the rate of 50 micrograms/hour for two hours to the 11 control subjects and 11 patients with liver cirrhosis (6 compensated and 5 decompensated). Basal plasma renin activity (PRA) in decompensated patients was significantly higher than those in control and compensated cirrhotics (P less than 0.01). Basal plasma aldosterone was also higher in decompensated than in control and compensated patients, but without significance. PGE1 had no virtual effect on PRA in control, but stimulated PRA in liver cirrhotics, in which statistical significance was only observed in decompensated (basal vs. one hour after PGE1: 2.4 +/- 0.9 ng/ml/min (mean +/- SE) vs. 6.9 +/- 2.1: P less than 0.025). The rate of renin release was significantly higher in compensated than in decompensated (327 +/- 50% vs. 143 +/- 26: P less than 0.05). Though PGE1 also increased plasma aldosterone in liver cirrhotics, statistical change was not seen. Fractional excretion of urinary sodium after PGE1 increased significantly in control (P less than 0.025), but not in liver cirrhotics. These results indicate that the renin angiotensin-aldosterone system is easily activated by PGE1 in patients with liver cirrhosis and further suggest that the sensitivity of this system in compensated is more augmented than in decompensated patients. PMID- 3042579 TI - Pancreatic polypeptide response to secretin in obesity: effects of glucose intolerance. AB - Pancreatic polypeptide (PP) may function as a regulator of satiety. Its secretion is impaired in certain animal models of obesity and the administration of PP may improve the hyperphagia and hyperinsulinism seen in these animals. In obese humans, decreased, normal or increased, basal and stimulated concentrations of PP in plasma have been reported. However the advent of diabetes confounds the picture since PP levels in diabetes are generally raised. We have therefore examined the PP responses to intravenous secretin, a known PP secretagogue, in 23 obese subjects, 12 with normal and 11 with abnormal glucose tolerance, and compared the results with those in 23 age and sex-matched healthy controls. The mean maximum PP level in obese subjects with normal glucose tolerance (98 +/- 13 pg/ml) was significantly less than that in normal subjects (218 +/- 23 pg/ml) but in obese subjects with abnormal glucose tolerance, it was significantly greater (578 +/- 115 pg/ml). Within each of the 3 study groups taken separately, PP response to secretin was not correlated with glucose or insulin levels, or with the degree of obesity. Thus, obesity per se appears to be associated with impaired PP responses, which may be masked by abnormalities in glucose tolerance. PMID- 3042581 TI - Ethanol-induced dissociation between whole pancreatic blood flow and islet blood flow in the rat. PMID- 3042582 TI - Immunogenotyping in Hodgkin's disease. AB - This review presents and discusses the immunogenotypic findings in 112 cases of Hodgkin's disease (HD) and eight Hodgkin's cell lines. Clonal rearrangements of the T cell receptor gamma and beta chain, as well as immunoglobulin heavy and light chain genes, are detected in the majority of nodular sclerosis and lymphocytic depletion subtypes. Together with the recent immunophenotypic data, these findings are in favour of the view that HD is a disease of activated lymphoid cells. Further investigations will be necessary to characterize the morphology and immunophenotype of the clonally rearranged cell population which seems not to be confined to the Sternberg-Reed and Hodgkin cell in every case. Prospective clinical studies including the genotype of HD cases have to be done in order to address the question of whether or not distinctive immunogenotypic profiles correlate with the clinical course of this lymphoproliferative disorder. PMID- 3042583 TI - Prognostic factors in decision-making in the clinical management of Hodgkin's disease. AB - At the beginning of this conference, Dr Ford asked whether or not Hodgkin's disease was a malignancy. Most physicians would agree that, regardless of whether this question can be answered on a molecular level, Hodgkin's disease certainly behaves like a malignancy, and clinically fits criteria necessary to call it one. It grows without control locally, comprising vital organs. It 'metastasizes', infiltrates organs, and causes organ dysfunction. Patients afflicted with the disease have a shortened life span without proper therapy, as demonstrated by DeVita in his original publication describing the benefits of MOPP chemotherapy (nitrogen mustard, vincristine, procarbazine, prednisone) (DeVita et al., 1970). However, it can progress very slowly, with 50 per cent of untreated patients with stage III or IV disease dead at one year, but 10 to 15 per cent alive at five years. Therefore, in its clinical behaviour, Hodgkin's disease certainly qualifies as a malignant disorder, despite the fact that we do not have the molecular means of calling it one. PMID- 3042584 TI - A review and interpretation of cytogenetic abnormalities identified in Hodgkin's disease. AB - The nature of the cell which gives origin to Hodgkin's disease remains unclear after years of intensive investigation. Cytogenetic data, which are very scarce in Hodgkin's disease, could contribute information which might help elucidate this problem. Review of our own data and others shows that the most frequent abnormalities in this disorder involve chromosomes 14q in 35 per cent of cases, 11q in 32 per cent, 6q in 32 per cent, and 8q in 18 per cent. The most common breakpoint in chromosome 14 occurred at 14q32 where the IgH chain genes reside thus suggesting that in these cases the cell of origin might be a B lymphocyte. The 11q and 6q structural abnormality have also been frequently seen in lymphoid disorders such as ALL and large cell lymphoma. Of interest also is the fact that in a certain type of childhood pre-B cell acute lymphoblastic leukemia which shows cytogenetic abnormalities on 11q23, aberrant myeloid and monocytic markers are seen. This suggests that in Hodgkin's disease a similar phenomenon might occur which could help to explain why the Reed-Sternberg cell expresses myeloid monocytic antigens such as Leu M-1. PMID- 3042585 TI - Advances in understanding the impact of stressful life events on health. AB - Throughout history, philosophers have debated the interaction of mind and body, but only in the last decade have psychiatrists and other health professionals begun to scrutinize how stressful life events may precipitate or contribute to the onset of illness. While traumatic experiences can profoundly affect physical and psychological well-being, their impact may be mediated by factors such as body chemistry and personality traits, which may predispose an individual to greater resilience or greater vulnerability to life stresses. This review covers research exploring the relationship between life stresses and illness, particularly the eating disorders and myocardial infarction, and describes efforts to develop objective instruments to assess the degree of stress associated with specific life events, a task known as life-events scaling. Avenues for future research are recommended. PMID- 3042586 TI - Inpatient and outpatient psychiatric services: substitutes or complements? AB - Patients at risk for psychiatric hospitalization make only limited use of outpatient services, although clinical research has demonstrated that outpatient treatments and home care can be as effective as inpatient psychiatric treatment in treating certain mental health problems. The substitution of ambulatory services for inpatient care has in part been limited because insurers have restricted outpatient mental health benefits to control utilization. The authors critically review evidence from the economic and clinical literatures to determine the extent to which outpatient psychiatric treatment substitutes or complements inpatient treatment. To encourage clinically effective substitution of outpatient for inpatient services, further research is needed to identify the patient populations for whom substitution is possible and the treatment and environmental conditions in which it is most likely to be successful. PMID- 3042587 TI - Electroconvulsive therapy for the elderly: a review. AB - Treatment of psychiatric disorders among the elderly is complicated by such factors as high incidence of medical illness and changes in drug metabolism; electroconvulsive therapy (ECT) is considered a reasonable treatment alternative for the elderly for several psychiatric syndromes. The authors review indications, complications, and precautions related to ECT for older patients. The primary indication is major depression; about 80 percent of elderly patients respond favorably. ECT appears less effective for depression secondary to dementia or somatization disorder. Although ECT is relatively safe for the elderly, up to one-third may experience a complication that interferes with treatment. Careful pre-ECT medical evaluation is essential, with special attention to cardiovascular factors and to concurrent medications. PMID- 3042588 TI - Counseling for HIV testing. AB - Recommendations are presented for counseling individuals who wish to know if they have been infected by the human immunodeficiency virus (HIV). Pretest counseling includes explaining the sensitivity and meaning of HIV antibody tests and the limits of confidentiality, assessing the patient's potential strengths, vulnerabilities, coping capacity, and supportive resources, and mutually deciding if the test is advisable. Populations that have been identified as at risk for HIV infection are listed, and four case vignettes that illustrate the complexity of the decision to be tested are provided. Posttest counseling includes notification of test results, reducing the immediate distress of seropositive patients, educating about how HIV is and is not spread, explaining methods to prevent transmission, advising the patient about who should be told the test results, and arranging follow-up care. PMID- 3042589 TI - Recent developments in brief psychotherapy. AB - The clinical effectiveness and many of the therapeutic mechanisms of brief psychotherapy have been validated through research studies in recent years, as indicated in this review of developments in the psychodynamic and interpersonal brief therapies. However, the literature shows considerable variation in methods for establishing time limits and in exclusion criteria. In a major shift in therapeutic practice, interpersonal rather than intrapsychic concepts are used to identify the focus of therapy, and systematic techniques for defining the therapeutic focus have been developed. The therapeutic alliance and its relation to outcome also have received substantial attention. Trends in service delivery clearly indicate increasing emphasis on brief psychotherapy, which must be conceptualized as a specific modality, not just as long-term therapy compressed. PMID- 3042590 TI - Characteristics of male spouse abusers consistent with personality disorders. AB - Researchers and clinicians have tended to minimize the role of psychological characteristics of male spouse abusers, which has restricted the selection of potentially appropriate treatment interventions. A review of clinical literature on psychological characteristics of male batterers, including some studies incorporating psychometric testing, suggests that many of the characteristics of batterers are consistent with DSM-III criteria for personality disorders. Several authors have defined subtypes of batterers, which can be associated with specific types of personality disorders. Treatment programs for male spouse abusers should address the specific problems presented by patients with personality disorders, including alcohol-abusing batterers, a particularly difficult group to treat. PMID- 3042592 TI - Hospitals dispute Medicare rules for liability care. PMID- 3042591 TI - Huge Medicare files to back outcomes research. PMID- 3042594 TI - HCFA will test revised Medicare cost reports. PMID- 3042596 TI - Health bill may help check drug misuse by elderly. PMID- 3042593 TI - Limitations of mortality data confirmed: studies. PMID- 3042595 TI - Pressured GPOs put hospitals in driver's seat. PMID- 3042597 TI - Morphologic studies of lymphocyte nuclei in follicular and diffuse mixed small- and large-cell (lymphocytic-histiocytic) lymphoma. AB - Twelve examples of mixed small- and large-cell lymphoma (eight follicular, one follicular and diffuse, and three diffuse) were investigated morphometrically using plastic-embedded tissue in order to study nuclear characteristics of lymphocyte populations in this form of non-Hodgkin's lymphoma (NHL) and to test morphologic bases for current NHL classification systems. This study illustrates that there are many inaccuracies, illusions, and misconceptions in the morphologic criteria currently used to classify mixed small- and large-cell lymphoma. A principal finding was that lymphocyte nuclear profiles in mixed-cell lymphomas tend to be smaller in size (P less than .005) and more irregular in shape (P = .0001) than the morphologically similar counterparts in germinal centers of lymph nodes with reactive hyperplasia. Intercase comparison of mixed small- and large-cell lymphomas revealed a considerable range of mean nuclear area values, some of which were within the size range of normal, small lymphocytes. At the magnifications used for morphometric assessment, a high proportion of lymphocyte nuclear profiles had shallow invaginations, but only a limited number of profiles (4% to 14%) had deep (cleaved) indentations. Contrary to current definitions for this subtype of NHL, lymphocytes with "small" nuclei had the same proportion of the nuclear diameter occupied by nuclear invaginations as lymphocytes with "large" nuclei and, in fact, mean nuclear invagination depth was shallower in "small" nuclei than in "large" nuclei. Furthermore, regardless of whether it is nuclear area or shape that is evaluated, lymphocytes in mixed cell lymphoma do not separate into two populations of small-cleaved and large noncleaved cells. Morphometry reveals that only four of the 12 examples of mixed small- and large-cell lymphoma had a proportion of the lymphocytes in the size range of fully transformed germinal center lymphocytes that exceeded 25%, and none of the cases approached 50% even though the population of lymphocyte nuclei appearing "transformed," and therefore "large," ranged from 28% to 57%. Such results indicate that the large, noncleaved and cleaved component, as seen in histologic sections of mixed small- and large-cell lymphoma, do not have nuclei of uniform size and many, in fact, are not actually large. The morphometric findings indicate reasons for the poor observer reproducibility in classifying this subtype of NHL. PMID- 3042598 TI - The caliber persistent artery of the stomach: a unifying approach to gastric aneurysm, Dieulafoy's lesion, and submucosal arterial malformation. AB - The caliber persistent artery of the stomach is the most dangerous form of gastrorrhagias; the overall lethality rate is 60.5%. A literature review aimed at completeness, a study of the hitherto largest case material (24 cases), and a comparative analysis of the bleeding and normal gastric arteries gave the following results: (1) the walls of the pathologic arteries are of normal structure; (2) as submucous arteries, they are of normal diameter; (3) they are attached to the mucosa by virtue of Wanke's musculoelastic mantle; (4) at the level of the muscularis mucosae, they are definitely oversized; (5) in the area of the linkage of the artery to the mucosa, a vulnerable mucosal spot is created; (6) the artery is accompanied by a vein of similar caliber; and (7) perforation of the vein takes place before that of the artery. In addition to surgical pathology and pathogenesis, the pitfalls of its diagnosis and treatment are also discussed. Caliber persistent artery of the stomach is much more common than the literature indicates, necessitating an increased awareness of the condition. PMID- 3042599 TI - A recombinant X chromosome in a short statured girl resulting from a maternal pericentric inversion. AB - A 7 3/4-year-old girl with short stature was found to have a recombinant (X),dup q chromosome resulting from an apparently unique pericentric inversion (X)(p11.2q26) present in her mother and maternal grandmother. The recombinant X chromosome was shown to be late replicating and the inversion X chromosome to be randomly inactivated. This appears to be only the eighth report (7 female, 1 male) of a recombinant resulting from an X pericentric inversion despite all diagnosed females having mild clinical abnormalities. Reasons for the rarity of such recombinant X chromosomes in man are examined. PMID- 3042600 TI - Oncogenes and cancer prognosis. PMID- 3042601 TI - The importance of dose intensity in the outcome of chemotherapy. PMID- 3042602 TI - Chemotherapy of urothelial tract cancer: Memorial Sloan-Kettering Cancer Center experience. PMID- 3042603 TI - Preservation of sexual function in the surgical treatment of prostatic cancer--an anatomic surgical approach. PMID- 3042605 TI - Immunotherapy of patients with advanced cancer using interleukin-2 alone or in combination with lymphokine activated killer cells. PMID- 3042604 TI - New reconstructive techniques in cancer surgery: the use of free and musculocutaneous flaps. PMID- 3042606 TI - Progress in the adjuvant therapy of large bowel cancer. PMID- 3042607 TI - Recent advances in the therapy of chronic myelogenous leukemia. PMID- 3042608 TI - Molecular biology of multidrug resistance in human cells. PMID- 3042610 TI - Gastrin-releasing peptide and other autocrine growth factors in lung cancer: pathogenetic and treatment implications. PMID- 3042609 TI - The use of hematopoietic growth and differentiation factors for bone marrow stimulation. PMID- 3042611 TI - Genetics and the etiology of solid tumors. PMID- 3042612 TI - [Pathogenesis of glomerulonephritis following streptococcal infection]. AB - Group A streptococci are responsible for the induction of serious pyoderma and pharyngitis in humans. The nonsuppurative sequelae "acute rheumatic fever" (ARF) and "acute poststreptococcal glomerulonephritis" (APGN) do very often occur with variable attack rates in epidemics with these microorganisms. At present, immune complex formation in the blood circulation system, in situ complex formation within the glomeruli, antibody cross-reactions between beta-hemolytic group A streptococci and human kidney tissue as well as tissue damage independent of specific antibodies are intensively discussed to explain the pathogenesis of acute poststreptococcal glomerulonephritis. However, the pathophysiological course of APGN has not yet been precisely identified. PMID- 3042614 TI - Stimulation of human and murine adherent cells by bacterial lipoprotein and synthetic lipopeptide analogues. AB - Lipoprotein from the outer membrane of Escherichia coli and its synthetically prepared N-terminal lipopeptide segments Pam3Cys-Ser-Ser-Asn-Ala and Pam3Cys-Ser, as well as lipoprotein from other Enterobacteriaceae, constitute potent polyclonal B lymphocyte activators. Here, we demonstrate that these compounds were also able to stimulate human and murine leukocytes: in murine macrophages, we could show the induction of interleukin 1 release by the mitogens, as measured in the thymocyte proliferation assay. Moreover, murine peritoneal exudate cells were stimulated to secrete prostaglandins E2 (PGE2) and F2 alpha (PGF2 alpha). The effect of Pam3Cys-Ser on the murine macrophage cell line P388D1 was also tested: the compound induced an increase in proliferation, as measured by a thymidine incorporation assay. In addition, the cell line could be induced to release IL 1 into the supernatant. Correspondingly, induction of IL 1 release could also be demonstrated in human mononuclear cells. Our results demonstrate that the two novel synthetic lipopeptides are potent stimulators for human monocytes and murine macrophages. These findings may be important for the elucidation of the role of these bacterial surface components in the course of bacterial infections. PMID- 3042613 TI - Biphasic radiation-sensitivity of the transplantation immunity for rejection of an allogeneic ascites tumor. AB - Transplantation immunity for second-set rejection of an allogeneic ascites tumor was induced by sensitizing mice with H-2-identical allogeneic spleen cells, and radiation-sensitivity of this immunity was studied. The immunity was not severely affected by 400 rads whole-body X-ray irradiation given at one day before the initial antigenic stimulation. In contrast, it was totally inactivated by 300-400 rads irradiation that was given one day before tumor challenge. An adoptive cell transfer experiment showed that alloreactive memory cells responsible for the immunity were unexpectedly highly radiosensitive. The immunity (memory), however, became resistant to 400 rads irradiation soon (one day) after challenge with the allogeneic tumor, and was resistant to 1500 rads for rejection of the tumor that was challenged at the second time when the initially challenged tumor was rejected. Corresponding to these observations, cells for allospecific CTL responses in peritoneal cavity of mice showed corresponding biphasic radiation sensitivity. PMID- 3042615 TI - A unique surface antigen on intraepithelial lymphocytes in the mouse. AB - This paper reports the discovery in the mouse of a new antigen found almost exclusively on the surface of lymphocytes residing in or immediately adjacent to the gut epithelium. The antigen was expressed by Lyt-2+ and L3T4+ cells but not by B cells or plasma cells and was present on almost all intraepithelial lymphocytes (IELs) in the gut. Only a very small proportion of cells in other lymphoid compartments expressed the antigen. Stimulation of IELs or other lymphocytes in vitro caused a decline in expression. Immunoprecipitation experiments showed the new antigen to be a molecular complex comprising two non covalently linked chains (175 kDa and 136 kDa) and minor components (27 kDa and 25 kDa). The function of this complex is unknown but its structure has certain features in common with that of cell adhesion molecules and extracellular matrix receptors of the 'integrin' supergene family. PMID- 3042616 TI - Immunodominance in T lymphocyte recognition. PMID- 3042617 TI - Effect of carbon and nitrogen on exoprotease synthesis in batch cultures of Bacillus licheniformis NCIM 2042. PMID- 3042618 TI - Influence of in vitro administered immune complexes on serum levels of complement and circulating immune complexes in Mycobacterium leprae infected mice. PMID- 3042619 TI - Escherichia coli that cause diarrhea. PMID- 3042620 TI - Neonatal enterovirus infection. PMID- 3042621 TI - Haemophilus influenzae type b vaccine. PMID- 3042623 TI - Treatment of tuberculosis in newborn infants and their mothers. PMID- 3042624 TI - Current status--meningococcal vaccine. PMID- 3042622 TI - Neonatal septicemia. PMID- 3042625 TI - Liposomes as drug delivery system in the treatment of infectious diseases. Potential applications and clinical experience. AB - Liposomes are microscopic, closed lipid vesicles able to entrap hydrophilic as well as lipophilic compounds. They constitute a versatile drug delivery system. When injected by the intravenous route, liposomes are taken up by macrophages in the liver and in the spleen. Investigation of several animal models of infections has shown that liposome-entrapped anti-infectious drugs are active against infections due to facultative intracellular bacteria, parasites such as Leishmania, viruses such as the one causing Rift valley fever. Liposomes of different lipid compositions, structures and sizes were used for intravenous administration of anti-infectious drugs without inducing toxicity in the tested animals. Clinical experience was obtained with two different liposomal preparations of amphotericin B in the treatment of systemic fungal diseases in cancer patients; these preparations were shown to be effective and very well tolerated. PMID- 3042626 TI - Treatment and prevention of pertussis by antimicrobial agents (Part II). AB - Suitable antimicrobials given during the catarrhal stage of whooping cough can attenuate the course of the disease. The efficacy of antibiotics administered prophylactically during the incubation period remains controversial but appears to be beneficial. Currently, erythromycin given for two weeks is the antibiotic of choice for pertussis. No treatment failures were observed with erythromycin estolate. Erythromycin ethylsuccinate and stearate must be given at high dosages (50-60 mg/kg/day) in order to achieve sufficient concentrations in the respiratory secretions. With ampicillin and amoxicillin treatment failures have been observed. The role of josamycin and co-trimoxazole in pertussis remains open. PMID- 3042628 TI - Osteomyelitis caused by Rhodococcus equi in a renal transplant recipient. AB - We report the first case of osteomyelitis due to Rhodococcus equi, which occurred in a renal transplant patient. Infection with this organism is rare and usually causes a distinct clinical syndrome resembling pulmonary tuberculosis. We investigated by time-kill curve analysis various antimicrobial combinations for in vitro efficacy. The literature is briefly reviewed, and aspects of diagnosis and therapy are discussed. PMID- 3042627 TI - Ofloxacin in the treatment of urinary tract infection in renal transplant recipients. AB - Forty-two renal transplant recipients suffering from complicated urinary tract infection (UTI) were treated orally with ofloxacin. Patients received an immunosuppressive treatment consisting of ciclosporin A and prednisolone (n = 39) or azathioprine and prednisolone (n = 3). Patients were routinely controlled for laboratory parameters before, during and after ofloxacin administration with special emphasis on nephrotoxicity and interaction of ofloxacin with the concomitantly administered immunosuppressives. Results show that ofloxacin does not impair renal graft function or interact with ciclosporin A or azathioprine even under long-term administration. It is effective and safe in the treatment of UTI in the renal transplant recipient. PMID- 3042630 TI - [Characteristics of nutrient utilization in cancer patients]. AB - The anabolic utilization of nutrients can be modified or impaired due to the influence of a malignant tumor in animals and in humans. Tumor-associated malnutrition as well as the side-effects of several antineoplastic regimens may induce mucosal atrophy, and by this the intestinal absorption of nutrients becomes impaired. Intensified nutrition therapy by oral, enteral or parenteral access is able to maintain or improve the nutritional status of tumor patients. The daily need of substrates can be enhanced as a consequence of the tumor induced stimulation of proteolysis, gluconeogenesis, hepatic protein synthesis, as well as of an impaired protein metabolism. In the last years, a lot of antimetabolites have been tested for antineoplastic treatment. So far, there is no drug of this kind known, which reduces tumor growth without harm for the tumor bearing host. PMID- 3042629 TI - Evaluation of protein metabolism in clinical practice. AB - The application of stable isotopes creates further possibilities for our understanding of the metabolism. New concepts especially for non-invasive diagnostic procedures could be developed. An important step in our research program was the performing of a 1-year experiment on a volunteer. On the basis of a 10-pool model we received a lot of informations. Based on this knowledge we developed 2 simplified methods for calculating whole body protein turnover. Knowing the problems with whole body protein calculations we intensified our intentions for determining the protein enrichment in organs and isolated cells (hepatocytes), estimating at the same time the precursor pool in the cells. Of special importance was an extensive study, wherein all those 15N-estimations were performed we are able to do up to now. We calculated whole body protein, but especially we studied the enrichment of the cellular protein fractions in hepatocytes, of plasmaproteins, and of the intracellular precursor pool. This study is the base for further tracer investigations. PMID- 3042631 TI - Digitalis and renal failure. PMID- 3042632 TI - Hypertonic hemodiafiltration. PMID- 3042633 TI - Regulation of pneumatic total artificial heart function: a review. PMID- 3042634 TI - Plasma exchange for the management of cyclosporin A-induced hypertriglyceridemia. AB - The authors report a case of hypertriglyceridemia complicating the course of a patient receiving cyclosporin A after bone marrow transplantation. When the patient was seen at the hemapheresis unit the clinical picture was characterized by headache, increasing visual and neurological disturbances. Plasma triglyceride level was 3215 mg/dl. Two plasma exchange sessions reduced triglycerides to 486 mg/dl and halted the disease progression. This may represent the first plasma exchange treatment of cyclosporin A-induced hypertriglyceridemia. PMID- 3042635 TI - Differential immunoreactivity and Ca2+-dependent degradation of vimentin in human fibroblasts and fibrosarcoma cells. AB - Immunostaining of normal human fibroblasts with a monoclonal antibody (MAb) (V22AC12) revealed typical cytoplasmic arrays of vimentin filaments in both mitotic and interphase cells. In human A8387 fibrosarcoma cells and SV40-virus transformed human fibroblasts, the same antibody showed positivity only in mitotic cells and in interphase cells only after treatment of the fixed cells with alkaline phosphatase. Upon immunoblotting with the MAb, an Mr 57,000 vimentin polypeptide was seen in normal fibroblasts. In fibrosarcoma cells the same polypeptide was revealed by this antibody only after treatment with alkaline phosphatase. The Mr 57,000 vimentin polypeptide was a major cytoskeletal protein in both fibroblasts and fibrosarcoma cells. Inclusion of Ca2+ into the cytoskeleton extraction medium brought about a somewhat increased degradation of vimentin in fibroblasts. In fibrosarcoma cells, such treatment caused a quantitative disappearance of the Mr 57,000 protein with a concomitant appearance of 3 distinct, low-molecular-weight degradation products in the detergent-soluble fraction. Another Ca2+-induced change in the polypeptide profile of fibrosarcoma cells was the disappearance of the Mr 240,000 non-erythroid alpha-spectrin and the concomitant appearance of a prominent Mr 140,000 degradation product. Inclusion of proteolysis inhibitors in the Ca2+-supplemented extraction medium inhibited degradation of both vimentin and alpha-spectrin polypeptides. The results suggest differences in the composition of the cytoskeletons of normal fibroblasts and fibrosarcoma cells, manifested in the differential Ca2+ susceptibility of vimentin and non-erythroid alpha-spectrin. Results with MAb V22AC12 suggest that differential phosphorylation of vimentin could account for at least part of this difference. PMID- 3042636 TI - Blood vessels and human essential hypertension. PMID- 3042637 TI - Body temperature in acute myocardial infarction and its relation to early intervention with metoprolol. AB - In a subsample of 223 patients participating in a double-blind trial with metoprolol in suspected acute myocardial infarction, body temperature during the first 5 days in hospital was recorded. Patients developing infarction had a mean temperature of 37.3 degrees C compared with 36.8 degrees C for those with no infarction (P less than 0.001). A positive association was observed between enzyme-estimated infarct size and body temperature (P less than 0.001). Patients given metoprolol had a mean temperature of 37.0 degrees C as compared with 37.2 degrees C in those given placebo (P = 0.03). The most marked difference between metoprolol and placebo was observed among those treated very early. We conclude that early treatment with metoprolol in suspected acute myocardial infarction appears to lower body temperature during the following days. This might reflect limitation of the infarct size. PMID- 3042638 TI - Immunohistochemical characterisation of Aschoff nodules and endomyocardial inflammatory infiltrates in left atrial appendages from patients with chronic rheumatic heart disease. AB - Fifty left atrial appendages collected fresh during closed mitral valvotomy in patients with chronic rheumatic heart disease, were analysed to determine the frequency of Aschoff nodules and characteristics of mononuclear inflammatory infiltration. Fifty-six percent of specimens demonstrated Aschoff nodules with no clinical or laboratory evidence of acute rheumatic activity in the patients undergoing surgery. Endomyocardial infiltration contained predominantly T cells and occasionally B cells. The relative proportions of T helper-inducer, T suppressor-cytotoxic lymphocytes and macrophages were 45.1 +/- 7.6, 23.5 +/- 4.8 and 29.3 +/- 9.6%, respectively. Frequent presence of Aschoff nodules and heavy mononuclear infiltrates in chronic rheumatic heart disease suggests a possibility of subclinical ongoing carditis. PMID- 3042640 TI - Effect of verapamil on basal and glucagon-dependent splanchnic glucose metabolism and insulin secretion in man. AB - To determine the effect of verapamil on basal and glucagon-induced changes in splanchnic glucose metabolism and insulin secretion, six healthy men were studied during a primed-continuous infusion of verapamil (2.5 micrograms/kg/min). After a basal period of 30 min, glucagon was infused at a rate of 12 ng/kg/min for an additional 60 min. Splanchnic exchange of glucose, lactate, alanine and beta hydroxy-butyrate, as well as splanchnic C-peptide release, reflecting insulin production rate, were determined by means of the liver vein catheter technique. Intraindividual control trials without verapamil were performed. Basal and glucagon-stimulated arterial glucose and insulin concentrations as well as insulin production rate were not significantly altered by verapamil, which also had no influence on basal splanchnic glucose output or on the glucagon-induced initial (0-30 min) increment in splanchnic glucose output. However, the gradual decline in splanchnic glucose output was delayed at 30-60 min (verapamil: 55.85 +/- 6.31 control: 39.73 +/- 5.6 mmol/30 min; mmol/30 min, mean, s.e.m., p less than 0.02). Splanchnic lactate-alanine- and beta-hydroxy-butyrate exchange were also unchanged by verapamil. We conclude that verapamil attenuates glucagon's evanescent effect on splanchnic glucose output without interfering with the exchange of gluconeogenic precursors or beta-hydroxy-butyrate or insulin production rate. PMID- 3042639 TI - Ribavirin influence on theophylline plasma levels in adult and children. AB - The pharmacokinetics of ribavirin and aminophylline (theophylline ethylenediamine) after oral administration in healthy adults and in children suffering from viral infections of the respiratory tract, with superimposition of bronchial asthma or asthmatic syndrome, was studied. By the prompt-release formulation of aminophylline the hematic peak is reached within the first 30 min (9.03 micrograms/ml) from intake, whereas by the sustained-release formulation the peak is reached only within the 5th hour (8.5 micrograms/ml). After administration of ribavirin no interferences with the clearance of theophylline both in adults and children were noted. PMID- 3042641 TI - Propofol, the newest induction agent of anesthesia. AB - Propofol is a rapidly acting intravenous anesthetic agent which has many advantageous kinetic properties explaining its usefulness by bolus dose for induction of anesthesia or for administration by continuous intravenous infusion. It is rapidly distributed in the body with a half-life of only around 2 min and has an efficient hepatic and extrahepatic clearance (total body clearance may exceed liver blood flow). Premedication has little effect on the already good induction characteristics of propofol. The incidence of cardiorespiratory depression appears to be higher than that of other induction agents, but, on the other hand, the absence of tachycardiac response prevents the increase in cardiac oxygen demands. In patients with cardiac disease, especially after high or repeated doses, propofol may be more depressant to the cardiovascular system than thiopentone resulting in imbalance of regional myocardial oxygen demand and supply. Recovery from propofol is rapid and clear-headed with almost no hangover effect. This makes it very suitable for out-patient anesthesia and for cardioversion. However, even with the new emulsion formulation of the drug, pain on injection is still a problem. With regard to a longer lasting combination anesthesia propofol remains an alternative to older induction agents. When given as a continuous intravenous infusion for total intravenous anesthesia or for sedation in intensive care unit propofol has shown little accumulation. Its clinical effects are predictable, consistent and recovery is rapid, independent of the dose given. Propofol has proved to be a useful induction agent regardless of the age of patients, but in the elderly there appears to exist a marked sensitivity to it. Up to now there is no evidence that propofol in emulsion drug form can produce allergic or anaphylactoid reaction more often than other induction agents in use and no severe hematological nor visceral toxicity have been reported. In the present situation, when althesin is not marketed any more due to a high frequency of anaphylactoid reactions and etomidate will have a limited use in clinical practice because of its blocking effect on adrenocortical function, propofol offers an important alternative anesthetic agent to thiopentone. PMID- 3042642 TI - Folklore therapeutic indigenous plants in periodontal disorders in India (review, experimental and clinical approach). AB - Though a number of plants and their parts are used for dental ailments among population in rural and urban areas of developing countries, in India however, the most common house-hold, road-side plants are mango (Mangifera indica), neem (Azadirachta indica; Melia azadirachta), ocimum (Ocimum basilicum), tea-dust (Camellia sinensis) and uncommonly murayya, i.e., currey leaf (Murayya koenigi) [Chopra et al. 1958, Kirtikar and Basu 1935, Nadakarni 1954, Satyavati 1984]. The leaves of these plants are folded and brushed (massage with teadust) against the teeth. Therefore, the present study is restricted only to the fleshy leaf extracts [Jindal et al. 1975] (except tea) of these plants inspite of certain limitations in the methodology and arbitrations in the microbial identification and isolation in the light of recent advances in folk dentistry. The investigation was carried out in two parts: 1) Experimental study: The efficacy of various dentifrices (commonly available in the market) and the potentiating effect of the leaf extract (LE) of the aforesaid indigenous plants when amalgamated with the tooth-paste against pathogens, were investigated. Further, the protection afforded by the said plant extracts (PE) over the conventional allopathic medicines on the human plaque cultures and gram negative bacteria from patients were studied. 2) Clinical study: The therapeutic effects of the said PE (individually) on clinical application among severely infected patients were examined. PMID- 3042643 TI - Ketoprofen vs etofenamate in a controlled double-blind study: evidence of topical effectiveness in soft tissue rheumatic pain. AB - The efficacy and safety of topical application of ketoprofen gel and etofenamate gel were studied in a controlled double-blind clinical trial. Thirty-six patients affected by inflammation of tendons, sheaths and bursae entered the study, and were treated for seven days. The parameters studied were: pain scale, Ritchie index, stiffness, pain on active and passive movement and functional capacity. The results showed that ketoprofen gel and etofenamate gel were able to induce remission of the inflammatory symptoms in soft tissue rheumatic pain. No side effects were detected in either group. PMID- 3042644 TI - Randomized double-blind study of flunarizine versus placebo in patients with chronic cerebrovascular disorders. AB - Chronic cerebrovascular disorders (CCVD), as defined by the 1980 Ad Hoc Committee in Paris, constitute both clinically and pathogenetically an extremely complex entity, characterized by a protean symptom pattern. The effects of a daily dose of 10 mg flunarizine orally on CCVD have been evaluated with a neuropsychological methodology during a three-month treatment period in a randomized double-blind study compared with a placebo. The results confirmed the effectiveness of the drug in the improvement of neurological, amnesic, attentive and behavioural symptoms, without evident side-effects even after a long-term treatment. PMID- 3042645 TI - "By the crowd they have been broken, by the crowd they shall be healed": the advent of group psychotherapy. PMID- 3042646 TI - Support groups for youth with the AIDS virus. PMID- 3042647 TI - The retrospective evaluation of competency to stand trial. PMID- 3042648 TI - The measurement of atherosclerotic peripheral arterial disease in epidemiological surveys. AB - In cardiovascular surveys, the WHO questionnaire on intermittent claudication has traditionally been used as a measure of atherosclerotic peripheral arterial disease of the lower limbs. But the questionnaire is of limited value because atherosclerotic disease is frequently asymptomatic. Research on the validity and reliability of the questionnaire and the simpler non-invasive tests of peripheral arterial disease are reviewed. These tests include measurement of the ankle brachial systolic pressure ratio, a treadmill exercise test, a reactive hyperaemia test, and assessment of toe-pulse reappearance time. The performance of these non-invasive tests is such that they should be used, in conjunction with the intermittent claudication questionnaire, in epidemiological studies of atherosclerotic peripheral arterial disease of the lower limbs. PMID- 3042649 TI - Excess risk of sickness and disease in bus drivers: a review and synthesis of epidemiological studies. AB - In an extensive search of available literature, 22 epidemiological studies that have examined health risks of bus drivers were identified. These studies focus on three main disease categories: (1) cardiovascular disease, including hypertension, (2) gastrointestinal illnesses, including peptic ulcer and digestive problems, and (3) musculoskeletal problems including back and neck pain. The studies consistently report that bus drivers have higher raes of mortality, morbidity, and absence due to illness when compared to employees from a wide range of other occupational groups. Increased disease rates have been found for drivers regardless of the use of different research methodologies, measurement techniques and comparison groups. When evaluating the impact of bias on the estimates of risk, it appears likely that findings are conservative: strong systematic selective factors have probably favoured the elimination of those in poorer health both at the time of entry into and exit from the job of bus driving and other sources of bias have most likely caused underestimations of risk. Nevertheless, there remain questions that need careful assessment before firm conclusions can be made about whether increased disease rates result from driving a bus. Such questions, coupled with the consistent findings of heightened risk of disease, make urban bus drivers an appropriate and promising occupational group in which to study further the potential adverse effects of the work environment on employee health. PMID- 3042650 TI - Cancer case-control studies with other cancers as controls. AB - Theoretical considerations concerning the use of other cancer patients as controls in cancer case-control studies are reviewed. Selection bias may be a problem in that some other cancers may be caused by the exposure under study biasing the odds ratio towards unity. Such bias is noted to be greatest with low prevalence exposures associated with high attributable risks for other cancers. However, it may be possible to identify selection bias with other cancer controls using census or other general population data. In addition, using other cancers as controls has important advantages with regard to recall and interviewer bias, which may be of unknown magnitude and direction when using general population controls. A further disadvantage of general population controls is that separate selection of decreased controls should usually be made for deceased cases, whereas a mixture of live and deceased controls can be expected when selecting other cancer patients as controls. Since there are also logistical and cost advantages in using other cancer patients as controls, this study design is likely to be used increasingly in the future, particularly in cancer registry settings. PMID- 3042651 TI - Evaluation and synthesis of health effects studies of communities surrounding arsenic producing industries. AB - Epidemiological studies designed to detect lung cancer risk and other health effects in communities surrounding arsenic-producing copper smelters were reviewed. The studies were about evenly divided in finding deleterious and 'beneficial' effects of arsenic. All of the studies had insufficient statistical power to detect the small increases in risk that may occur. Even the most powerful studies were not designed to detect relative risks less than about 1.2 and the majority of the studies had little power to detect risks under 2.0. Confidence intervals for the relative risks from these studies were not very useful in putting an upper bound on adverse effects of arsenic. Sources of bias and other difficulties with community health studies are also discussed. We argue that these studies may be a good and economical first investigation but, due to a lack of power, null findings do not rule out the possibility of excess risks that may be significant from a public health viewpoint. PMID- 3042652 TI - Human pituitary growth hormone (hGH) and Creutzfeldt-Jakob disease: results of an epidemiological survey in France, 1986. AB - An epidemiological inquiry has been done in France after the notification in the USA and England of four cases of Creutzfeldt-Jakob disease in patients previously treated with hGH. Between 1959, when hGH treatment in France was started, and August 1985, the date the survey began, 1698 patients were registered for treatment. Current information (less than three months old) was obtained for 1620 patients (95.4%). Death was reported in 31 patients, but none could be related to Creutzfeldt-Jakob or similar disease. Pathological events were observed in 213 living patients (13.1%). Among them, four were diseases classified as possibly related to a viral infection. The first case had acute lymphoid leukaemia; the second case had polyradiculoneuritis associated with hepatitis. In both cases the disease resolved completely. Two other patients had acute encephalitis which started less than two years after the onset of treatment and which resolved spontaneously. Even though the acute evolution and the spontaneous clinical recovery are not consistent with Creutzfeldt-Jakob disease, a relationship with hGH therapy could not be completely excluded. Finally, five treated children had later malignancies which raises the question of the long-term secondary effects of hGH upon cellular proliferation. PMID- 3042653 TI - Immunogenicity and safety of measles vaccine in ill African children. AB - A concurrent prospective study was conducted in Rwanda to compare the immunogenicity and safety of live, attenuated measles vaccine in ill and well children. Five hundred and eighteen children aged 8 to 19 months were selected from children attending the acute care and immunization services of two clinics. Two hundred and sixty-seven ill children and 251 well children were enrolled and examined. Serological tests were performed on blood samples obtained before and 40 days after measles immunization. Among the 208 ill children and 215 well children who were seronegative at baseline and had unequivocal follow-up serological results, seroconversion rates were 81% and 80%, respectively. Side effects were modest and were equally frequent in the two study groups (15.4% among ill children versus 15.1% among well children). These results support a change in measles immunization policy in developing countries with respect to immunization of children with acute illnesses. Such a change would make a great contribution to decreasing the enormous burden of measles in the developing world through increased immunization coverage. PMID- 3042655 TI - Notes on Sigmund Freud's 'Analysis Terminable and Interminable'. AB - The author suggests that 'Analysis terminable and interminable' should be considered in the context of its particular background of political and scientific history as well as of Freud's personal biographical circumstances. It should not be considered only as a technical discussion. The author looks at Freud's own statements about the paper, together with those of contemporary (Fenichel) and later commentators. PMID- 3042656 TI - An historical, biographical, literary, and clinical consideration of Freud's 'Analysis Terminable and Interminable' on its fiftieth birthday. AB - Freud's paper is re-examined and placed in biographical and historical context, Moses and monotheism (1934-1938) is treated as a parallel text. Freud's ironic scepticism is viewed against the background of Thomas Mann's 1936 idealization of psychoanalysis. Freud was a sick and dying man in 1937 with every reason for personal pessimism. The political setting was the triumph of fascism in Austria and the impending seizure of Austria by Hitler. Freud fantasized the protection of psychoanalysis by the Roman Catholic Church. The paper is a polemic against the theories of Ferenczi, Adler, and Rank. Contemporaneous defence of prophylactic analysis by Fenichel is reviewed. Freud's political judgement of the moral equivalence of Social Democracy and fascism is critiqued. Freud's injunction to periodic re-analysis and his vision of analysis as 'unending' is supported. The possibility of intergenerational prophylaxis is presented. PMID- 3042657 TI - Development terminable and interminable. II. Recent psychoanalytic theory and therapeutic considerations. AB - This is the second part of a paper previously published in the Journal. It reviews how the formulations proposed in Part 1 are not only consistent with interdisciplinary research findings from developmental psychobiology and from infant observations, but are also consistent with a range of clinical theories of psychoanalysis. The basic motives of infancy and the motivational structures arising from early relationship development are the subject of increasing attention in psychoanalytic treatment. Aspects of the working alliance, both in the beginning and termination phases include considerations of emotional availability, the use of positive emotions, as well as the relationship 'match' between patient and analyst. These may determine outcome and are deemed worthy of further inquiry. PMID- 3042654 TI - One psychoanalysis or many? AB - This paper describes the historic development of psychoanalysis, the singular product of the genius of one man, Sigmund Freud, and which he made such strenuous efforts through his lifetime to maintain as a unified enterprise, defining out dissidents (like Adler, Jung, etc.), into what has become in the almost half century since his death a science and a discipline characterized by an increasing diversity or pluralism, of theoretical perspectives, of linguistic and thought conventions, and of distinctive regional, cultural, and language emphases. I discuss both the understanding of this theoretical diversity and what, in the face of it, holds us together as common adherents of a shared psychoanalytic science and profession. My thesis is that what unites us is our shared focus on the clinical interactions in our consulting rooms, the phenomena encompassed by the 'present unconscious' (the Sandlers) or the 'clinical theory' (George Klein). When we look beyond that to an explanatory structure within which to conceptualize the genetic-developmental process, normal and abnormal mental functioning, psychopathology and its reversal (cure) i.e. the realm of the 'past unconscious' or the 'general theory', we posit our various theoretical, linguistic or thought perspectives, i.e.--at this stage of our historical development--our metaphors or our various explanatory symbolisms. Some implications of this viewing of our various theoretical perspectives in psychoanalysis (ego psychological, object-relational, self-psychological, Kleinian, Bionian, Lacanian, etc.) as but metaphoric expressions, are reviewed. PMID- 3042659 TI - The epidemiology of sports-related ocular trauma. PMID- 3042660 TI - Ocular injury due to light toxicity. PMID- 3042661 TI - Molecular genetics of Duchenne and Becker muscular dystrophy. PMID- 3042658 TI - Inhibition of prolactin release by luteinizing hormone-releasing hormone in Tourette's syndrome. PMID- 3042662 TI - Neurotoxin-binding site on the acetylcholine receptor. PMID- 3042663 TI - Pathobiology of neuronal storage disease. PMID- 3042664 TI - Thalamic amnesia: clinical and experimental aspects. PMID- 3042665 TI - Critical notes on the specificity of drugs in the study of metabolism and functions of brain monoamines. PMID- 3042666 TI - Retinal transplants and optic nerve bridges: possible strategies for visual recovery as a result of trauma or disease. AB - From the review of the current literature it is quite evident that some exciting prospects are on the horizon which will help to better explain the development and functioning of the visual system. In addition, the new technology of CNS tissue grafting coupled to other newly emerging technologies (i.e., microsurgical, microinjection, and micromanipulative techniques coupled with our knowledge of immunosuppressive methods) will allow for a realistic approach in exploring possible strategies for visual recovery as a result of trauma or disease within the near future. One specific area of research that hopefully will emerge from this new body of knowledge comes from the realization that at the present time there is no effective therapy for practically all types of hereditary retinal degenerative disorders in man. It would seem most appropriate to take advantage of the new neuronal transplantation technology mentioned in this article and the availability of hereditary retinal degeneration models in the hope of developing new methods for a therapeutic approach to this problem. Such an approach could involve replacing the abnormal, absent, and/or lost host retinal cells with tissue from healthy donors by means of a grafting technique with the goal of arresting and/or reversing the disease process. Of course, this is but one example of the many challenges in this area of research which increasingly appear to be within our grasp. PMID- 3042667 TI - Schizophrenia: instability in norepinephrine, serotonin, and gamma-aminobutyric acid systems. PMID- 3042670 TI - [Definition and diagnosis of alcoholism]. PMID- 3042668 TI - Changes of clinical biochemical values after bilateral nephrectomy and allotransplantation of the kidney in pigs. AB - In a series of 10 healthy pigs, the normal ranges of the following biochemical parameters in blood serum were assessed: creatinine, urea, mineralogram (Na, K, Cl), uric acid, bilirubin, total protein, cholesterol, blood lipid level, thymol turbidity reaction and activities of alkaline phosphatase, alpha-amylase and transaminases. The changes of these values were followed up in two pigs after bilateral nephrectomy and in 18 pigs subjected to orthotopic allotransplantation of the kidneys. The examination confirmed that creatinine and urea are the most important values with a prognostic impact. However, other biochemical values signalize also renal dysfunction. In the mineralogram, values of Na and K ions are important indicators. Deterioration of renal functions was associated as a rule with a decline of alpha-amylase and alkaline phosphatase. The other examinations did not display any marked changes during renal damage. PMID- 3042671 TI - [Psychological changes in alcohol abuse]. PMID- 3042669 TI - Lymphocyte subpopulations in mesangial IgA glomerulonephritis. AB - Lymphocyte subpopulations in thirty patients with IgA glomerulonephritis and in twenty-four healthy persons were identified using specific monoclonal antibodies. Decreased OKT11+ cells (37.83 +/- 13.34, p less than 0.01) without any changes of OKT4+ cells, but increased OKT8+ cells (25.89 +/- 6.70, p less than 0.01) in patients with IgA glomerulonephritis, as compared to a control group, were found. The OKT4+:OKT8+ ratio was decreased in the patients (2.18 vs. 2.47 for the control group). The monocytes were increased (19.26 +/- 6.63 vs. 2.47 +/- 3.28). No statistically significant correlations between abnormal T cells or monocytes and blood pressure, serum creatinine, erythrocyte sedimentation rate and serum levels of IgA were found. It is concluded that no clear-cut correlation could be demonstrated between in vitro and clinical findings. PMID- 3042672 TI - [Metabolism of alcohol. Significance and metabolic effects]. PMID- 3042673 TI - [Epidemiology and prevention of alcoholism in West Germany]. PMID- 3042675 TI - [Induction and increase of cardiac arrhythmia by anti-arrhythmia agents]. PMID- 3042676 TI - [Faulty use of antibiotics and the development of resistance]. PMID- 3042674 TI - [Organ damage caused by alcohol]. PMID- 3042677 TI - [HELLP (hemolysis, elevated liver enzymes, low platelet count) syndrome as a special form of severe EPH gestosis]. PMID- 3042678 TI - [Fever with vesicular exanthema and pharyngitis in a 20-year-old patient (Mycoplasma pneumoniae associated Stevens-Johnson syndrome)]. PMID- 3042680 TI - [Living with AIDS. Position and perspectives of AIDS control in West Germany--a memorandum of the German AIDS Service]. PMID- 3042679 TI - [Hemolysis, methemoglobinemia and renal failure in a 30-year-old patient]. PMID- 3042681 TI - [Insulin receptor defects as a cause of disease]. PMID- 3042682 TI - [Disorders of receptor function as a pathogenetic principle in myasthenia gravis]. PMID- 3042683 TI - [Pathogenetic significance of receptors in bronchial asthma]. PMID- 3042684 TI - Radioimmunodiagnosis through imaging. Prospects for tumor imaging using labeled monoclonal antibodies. PMID- 3042685 TI - Lesion detectability in ultrasonic computed tomography of symptomatic breast patients. AB - From 95 subjects imaged with both speed of sound and attenuation ultrasonic computed tomography (UCT), analyses were performed on 40 cases for which unequivocal clinical diagnoses were available for correlation. This paper describes the UCT image characteristics and addresses the hypothesis that carcinomas and other lesions can be detected and localized by means of simple visual criteria or lesion characteristics that are quantitative relative to those of other breast tissues in the same patient. The most useful within-patient criterion was selection of the solid mass with the highest speed of sound in either breast (12 of 12 carcinomas). Architectural asymmetry between breasts in the three types of images was a significant contributing factor in visual image interpretation in seven of the eight cancer patients in whom there were comparable images of both breasts. Solid masses were discriminated by attenuation coefficient and pulse echo criteria. Our results did not substantiate the hypothesis that the average speed of sound throughout the cancer containing breast would be higher than in the contralateral breast. These results are better than might be expected from pulse echo imaging alone on this population. However, clinical implementation probably should be deferred until the technique is made more convenient and less expensive, or more accurate with a greater promise for diagnosis of minimal cancers. PMID- 3042686 TI - Tolerability of hypertonic and isotonic contrast media injected intravenously. A comparative study in the dog. AB - We examined the use of isotonic and hypertonic contrast media injected intravenously in the dog from the standpoint of cardiovascular tolerance after right atrial injections performed at 2.56 and 5.12 g I/second. The parameters measured were lead II of the electrocardiograph, heart rate, pulmonary and abdominal arterial pressure, and aortic flow. Three contrast media, ioxitalamate, ioxaglate, and iopamidol (two ionic and one nonionic), were compared, either concentrated (32% iodine) or dilute and isotonic with plasma (ioxaglate 160 mg I/mL and iopamidol 128 mg I/mL). At an injection rate of 5.12 g I/second, iopamidol-128 showed lower electrophysiologic tolerability and caused a higher increase in aortic flow than ioxitalamate 160 or ioxaglate 160. These effects may explain the lower radiographic efficacy observed with iopamidol-128 in previous digital subtraction angiography studies. PMID- 3042687 TI - Analysis of the methylation pattern of c-Ha-ras oncogene in human prostatic cancer. AB - To determine the methylation pattern of c-Ha-ras oncogene in prostatic tissue and to identify possible changes of methylation associated with cancer, high molecular weight DNA was extracted from 7 normal and 6 carcinomatous human prostates. Analysis of the samples was performed by cleaving DNA with the restriction endonucleases Msp I, Hpa II and Cfo I, and by Southern hybridizing the DNA digests with the 32P-labelled c-Ha-ras (pT24-C3) probe. Several discrete fragments were obtained with Hpa II and Cfo I digestion while the Msp I pattern showed fewer and smaller bands. Therefore, c-Ha-ras appears to be partially methylated. While a considerable polymorphism of the sequence 5'-CCGG-3' was observed at several Msp I sites in all cases, no significant differences could be evidenced in the methylation patterns of normal and neoplastic prostatic DNA samples extracted and purified from each patient. PMID- 3042688 TI - Effect of the triterpenoid fraction of Centella asiatica on macromolecules of the connective matrix in human skin fibroblast cultures. AB - The mechanism of action of the total triterpenoid fraction extracted from Centella Asiatica (TTFCA) was evaluated using human skin fibroblasts cultures as the experimental system. In particular its influence on the biosynthesis of collagen, fibronectin and proteoglycans was considered. The presence of TTFCA (25 micrograms/ml) does not seem to affect cell proliferation, total protein synthesis or the biosynthesis of proteoglycans in a significant way. A statistically important increase was observed in the percentage of collagen and, as revealed by immunofluorescence measurements, in cell layer fibronectin. This effect on collagen and fibronectin may help to explain the action of TTFCA in promoting wound healing, and suggests an interesting working hypothesis for its action on basal endothelia. PMID- 3042689 TI - Interstitial irradiation therapy of supratentorial gliomas by stereotaxic technique. Long term results. AB - We report the long term results of interstitial irradiation therapy in patients suffering from malignant supratentorial gliomas. The radioisotopes implanted by stereotaxic technique were Au198 grains in solid tumors and Y90 colloidal solution in a cystic tumor. The therapy was always well tolerated. Minimum survival time after implantation was 9 months, maximum exceeded 52 months. In all cases interstitial irradiation therapy solved or drastically diminished the severe secondary epilepsy from which all the patients suffered. PMID- 3042690 TI - Complicated migraine: case report. AB - The occurrence of long lasting focal neurological deficit as a complication of migraine is well known. A high incidence of C.T. scan abnormalities have been seen in subjects affected by severe complicated migraine; in some cases such lesions had the aspect of cerebral infarction. In this report we will relate case of a 29 year old woman with complicated migraine and multifocal suffering within the area of the hind brain circulation. The hypodense area we found with C.T. in the left cerebellar hemisphere and the reversibility of this lesion could support the hypothesis of focal edema in our case. PMID- 3042691 TI - Autosomal dominant carpal tunnel syndrome in a Karaite family. AB - A clinical examination and a history of 64 members belonging to four generations of a Karaite kindred revealed 11 patients with carpal tunnel syndrome (CTS). Bilateral involvement and early age of onset are salient features. The mutation may be traced in four generations. Hereditary primary CTS may be more prevalent than hitherto suspected. PMID- 3042692 TI - Cerebrohepatorenal syndrome of Zellweger: a peroxisomal deficiency disorder. Case report and review. AB - The cerebrohepatorenal syndrome of Zellweger (CHRS) is a rare, fatal disorder in newborn infants. Recent research points to a primary absence of tissue peroxisomes, with resulting biochemical defects, as the basic pathology of the condition. We report on an infant with classic neurological and dysmorphic features of CHRS. Increased serum levels of pipecolic acid, bile acid precursors and very-long-chain fatty acids (VLCFA) together with total histological absence of liver peroxisomes confirmed the diagnosis. To our knowledge, this is the first case in Israel to be fully documented by biochemical and ultrastructural techniques. A high index of suspicion is essential among clinicians if further cases are not to be overlooked. PMID- 3042694 TI - Use of total lymphoid irradiation (TLI) for the treatment of autoimmune disorders. PMID- 3042693 TI - Autoantibodies against nuclear ribonucleoprotein (RNP) complexes. PMID- 3042695 TI - Osseointegrated jawbone implants for permanent dental and maxillofacial reconstruction. PMID- 3042696 TI - Migraine headaches, migraine equivalents and anti-smooth muscle antibodies. PMID- 3042697 TI - Aspartame and headache. PMID- 3042699 TI - [Motility disorders of the esophagus in progressive systemic scleroderma. Pathophysiology, diagnosis and therapy]. AB - Gastrointestinal manifestations of collagen diseases are frequent. In progressive systemic sclerosis esophageal involvement is found in 60% of cases and is thus the main gastrointestinal complication. Atrophy of the smooth muscle and fibrotic degeneration of the distal esophagus result in progressive motility disorders which may cause severe reflux esophagitis with typical consequences, such as stenosis and strictures. Manometry and cinematography are basic diagnostic procedures. Esophagoscopy and long-term pH-monitoring are most useful for evaluating the degree of esophageal involvement. The severity of sclerodermatous motility disorders should be classified according to a modification of the Garrett scale, which is particularly recommended for determining the further prognosis and therapeutic approach. Esophageal involvement of grades I and II should be treated conservatively, whereas grade III is a clear indication for surgical therapy. The original Nissen type of fundoplication or distal gastric resection with Roux-en-Y anastonosis are the methods of choice. PMID- 3042698 TI - [Disseminated superficial actinic porokeratosis with Bowen's disease]. AB - While there have been several reports about the development of malignant neoplasms in porokeratosis of Mibelli and porokeratosis plantaris, palmaris et disseminata, only one case of squamous cell carcinoma in disseminated superficial actinic porokeratosis (DSAP) has been documented. A review is presented of the variants of porokeratosis that have so far been described in the literature as well as the differential diagnosis, genetic aspects and therapy of DSAP. The first case of DSAP with Bowen's disease is presented. PMID- 3042700 TI - [Gemmangioma. Case report and review of the literature]. AB - A benign gemmangioma with a diameter of 1 cm was found on the mons pubis of a 56 year-old woman. Gemmangiomas are rare tumours that occur in different locations. They have their origin in pluripotentional angioblastic cells and develop typical immature capillaries in a mesenchymal tumour stroma with few cells. The great variability and uncertain nature of the tumours are caused by the pluripotential capacity of the tumour cells. A review is presented of the cases reported in the literature since the first description by Orsos in 1932. PMID- 3042701 TI - [The Bloch era as reflected in the moulages collection at the Zurich Dermatological University Clinic 1917-1933]. AB - The collection of moulages at the Department of Dermatology, Zurich University Hospital, is well preserved and still offers the possibility of following the scientific interests of it's founder, Bruno Bloch. These are presented here together with a short biography of Bloch. PMID- 3042702 TI - [Comments on the paper by R. Rudlinger and D. Neumann-Haefelin: Cytomegaloviruses and Kaposi sarcoma]. PMID- 3042703 TI - [Antimycotic susceptibility testing of yeasts important in dermato-venereology: methods, results and clinical relevance]. AB - During the last decades many different methods have been devised for testing the antimicrobial susceptibility of yeasts in vitro. These methods range from agar diffusion and agar dilution to macro- and micro-broth-dilution methods. Although the ideal method has not yet been found, interest is currently focussed on micro dilution tests. Yeasts that cause disease in man are not generally susceptible to the most frequently used antimycotics. This is especially true of 5 fluorocytosine, but strains resistant to ketoconazole, nystatin and amphotericin B have also been found. So far, many of the data obtained from in vitro studies of the antimicrobial susceptibility of yeasts have been found not to be closely correlated with clinical outcome. Most recently this situation has changed greatly, however, and this seems to be due to a large extent to new test procedures. PMID- 3042705 TI - [Congenital telangiectatic erythema (Bloom syndrome)]. AB - "Congenital telangiectatic erythema" (Bloom's syndrome) is very rare; it is linked to a group of hereditary diseases and a common feature is cellular hypersensitivity to a variety of physical or chemical agents. The mode of transmission is autosomal-recessive. Characteristic criteria for Bloom syndrome are: consanguinity of the parents; androtropia; low birth weight, short stature (proportional); and persistent telangiectatic erythema in sun-exposed areas, sometimes with blistering. The syndrome is also associated with a facultative lack of antibodies. Of great importance is the high leukaemia morbidity among individuals with this syndrome; chromosomal aberrations and breakages play a significant role. Histological changes comprise an increase in dilated vessels in the upper dermis and damage and loss of elastic fibers. We give a review of Bloom's syndrome and present a case report. PMID- 3042704 TI - [Acquired epidermolysis bullosa. A clinico-pathologic study]. AB - Epidermolysis bullosa acquisita is an immunologically mediated mechano-bullous dermatosis presenting with a variety of clinical appearances. In mild cases the skin lesions, serous blisters, erosions and scars are restricted to the extremities. Scarring alopecia, dystrophy and even loss of nails as well as extensive erosions of mucous membranes may complicate severe cases. The characteristic histopathological feature is a subepidermal blister, which is located within the dermis by electron microscopy. Linear deposits of IgG and C3 along the basement membrane zone have been found on the dermal side of the lamina densa by immunoelectron microscopy. This close anatomical relationship with anchoring fibrils strongly suggests a functional disturbance of these anatomical structures, leading to dermolytic blistering. Treatment with sulphones, in combination with cortico-steroids in the more severe cases, has resulted in long term improvement in two of our three patients. PMID- 3042707 TI - [Comments on the paper by K. Bork: Psoriasis and bullous pemphigoid]. PMID- 3042706 TI - [Spontaneous manifestation and regression of a Kaposi's sarcoma under cyclosporin A therapy]. AB - A case of Kaposi sarcoma is reported in a 40-year-old Turk 3 months after a kidney transplantation under immunosuppression with cyclosporin A and methylprednisolone. After reduction of immunosuppression, there was complete regression of the sarcoma and the kidney transplant functioned correctly. PMID- 3042708 TI - [Early detection of malignant melanoma of the skin]. PMID- 3042710 TI - [Amputation surgery. General aspects and principles]. PMID- 3042709 TI - Access to ambulatory care for poor persons. AB - Studies conducted during the 1970s have reported conflicting results concerning whether differences in use of physician services between poor and nonpoor persons have been eliminated. Using a sample of 92,737 persons from the 1982 National Health Interview Survey and a refined analytic method, this study reexamines utilization of ambulatory care services by persons above and below the poverty level. Before adjusting for health status, no differences were apparent in rates of physician contacts for persons above and below the poverty level. After adjusting for health status, persons below poverty were shown to have significantly fewer physician contacts than persons above poverty. Multivariate analysis revealed that Medicaid coverage can effectively counter the depressing effects of poverty on use of physician services. However, only 34 percent of the noninstitutionalized population below poverty level had Medicaid coverage in 1982. Public policy implications concerning Medicaid eligibility criteria are discussed. PMID- 3042712 TI - [Disarticulation of the knee and the Gritti amputation]. PMID- 3042711 TI - [Amputation at mid-thigh]. PMID- 3042713 TI - [Ruptures and perforations of the thoracic and abdominal esophagus. Incidence and treatment]. PMID- 3042714 TI - [Surgical ulcer prevention in transplantation surgery]. PMID- 3042717 TI - Interzonal and intrazonal heterogeneities in the renin status of the preglomerular arterioles in five species. AB - In five species (mouse, rat, rabbit, rhesus monkey and man) the renin status of the preglomerular arterioles was examined using two immunohistochemical methods: the measurement of the renin-positive portion of the vessels, reflecting the respective number of granulated cells, and the semiquantitative assessment of the renin concentration in the juxtaglomerular epithelioid cells with antibody dilution series. The main objective of the study was to compare the interzonal with the intrazonal internephron heterogeneities, i.e. the differences between the average renin status of the preglomerular arterioles in the superficial, intermediate and juxtamedullar cortex with the differences between the renin status of the individual afferent arterioles in one and the same cortex region. In contrast to small interzonal heterogeneities, substantial intrazonal differences in the renin status of the corresponding nephrons were found. PMID- 3042716 TI - In situ immunocytochemical staining of cell colonies growing in plasma clot. AB - The procedures involving the growth of cell colonies in semi-solid media, such as methyl cellulose or agar, provide a score of colony-forming-units (CFUs) by means of morphology, and allow the application of cytochemistry. However, a better characterization of the growing cells by employing monoclonal antibodies is impaired by the medium itself. Plasma clot is a possible alternative, allowing immunofluorescence as well as immunoenzymatic techniques. We have developed a staining procedure which can be performed using both peroxidase- or alkaline phosphatase-conjugated reagents; the colonies, growing in plasma clot, can be stained in situ, without transferring the cells. In this paper we report on the study of six different cell lines stained by immunocytochemical techniques with appropriate monoclonal antibodies. PMID- 3042718 TI - What do we know today about welding-fume effects on the respiratory system? PMID- 3042719 TI - Radiosurgery for arteriovenous malformations of the brain using a standard linear accelerator: rationale and technique. AB - We have recently initiated a program for irradiating small, unresectable arteriovenous malformations (AVM's) in the brain. The treatments are delivered using a modified and carefully calibrated 6 MV linac. We are using high, single doses (15 to 25 Gy) with a goal of sclerosing the vessels and preventing hemorrhages. This technique, radiosurgery, is somewhat controversial in the radiotherapy community. Since the treatment is given in a single sitting, rather than in the more conventional pattern of multiple small daily fractions, there is some concern about late radiation damage to the normal brain tissue. However an extensive review of the literature leads us to the conclusion that if a technique is used that keeps the volume irradiated to high dose small, radiosurgery is a safe and efficacious treatment for small (less than 2.5 cm) AVM's. To decrease the risk of necrosis of normal brain tissue, it is important to confine the high dose region as tightly as possible to the target volume. Precise target localization and patient immobilization is achieved using a stereotactic head frame which is used during angiography, CT scanning, and during the radiation treatment. This minimizes the margin of safety that must be added to the target volume for errors in localization and set-up. The treatment is delivered using multiple noncoplanar arcs, with small, sharp edged X ray beams, and with the center of the AVM at isocenter. This produces a rapid dropoff of dose beyond the target volume. Early results in our first few patients are encouraging. PMID- 3042715 TI - Vitamin D--soltriol the heliogenic steroid hormone: somatotrophic activator and modulator. Discoveries from histochemical studies lead to new concepts. AB - Evidence from autoradiographic studies with 3H 1,25(OH)2 vitamin D3 (soltriol) about its many sites of nuclear binding and multiple actions suggests that the traditional view of "vitamin D and calcium" is too limited and requires modification. A new concept has been developed which proposes that the skin derived hormone of sunshine, soltriol, is a somatotrophic activator and modulator that affects all vital systems. Regulation of calcium homeostasis is only one of its many actions. Target tissues for soltriol include not only bone, intestine and kidney, but also brain, spinal cord, pituitary, thyroid, endocrine pancreas, adrenal medulla, enteroendocrine cells, thymus, and male and female reproductive organs. Accordingly, actions of soltriol involve effects on autonomic and endocrine regulation with changes in tissue and blood hormone levels, innervation of skeletal muscle, immune and stress response, digestion, blood formation, fertility, pregnancy and lactation, general energy metabolism, mental processes and mood, and others. The skin-mediated transduction of short-wave sunlight induces a purposeful modulation of growth, reproduction and other biological activities in tune with the conditions of the sun cycle and season. Synthesis and actions of vitamin D3-soltriol are dependent not only on the amount of sunlight, but also on the availability of precursor in the skin and access of sunlight, the rate of hydroxylation in liver and kidney, and the modulation of these events by the endocrine status, in particular growth and reproduction. A concept of a five level control of soltriol synthesis is proposed, in which the hydroxylation steps provide for a sensitive tuning. Relationships between the heliogenic skin-derived hormonal system and the helioprivic pineal-derived hormonal system are recognized and a comprehensive concept of the "endocrinology of sunlight and darkness" is pointed out. PMID- 3042720 TI - From manual to 3-D computerized treatment planning for 125-I stereotactic brain implants. AB - Aspects of planning for the treatment of high grade primary or recurrent brain tumors with stereotactically placed catheters afterloaded with high activity 125 I seeds are discussed. At our institution, planning has evolved from a simple manual process, which assumed geometric symmetry, through a more advanced manual process, that took advantage of certain mechanical properties of the stereotactic frame used, into a sophisticated, computerized planning approach that includes optimization of the source distribution and 3-D displays. Use of the simple manual method is limited to the rare situations where target volumes are quite regular in shape. The advanced manual method provides some customization for irregularly shaped volumes, but is slow and tedious to implement. The interactive, computerized approach permits identification of target volumes directly on CT slices, reconstructions in arbitrary planes, and optimization of catheter placement, source separation along each catheter, and selection of source strengths from an available inventory. A multi-format display feature which includes a probe's eye view perspective is provided to aid in planning. Integral dose-volume histograms for the target volume point out the advantages in using sophisticated, 3-D, computerized planning systems for these implants. PMID- 3042721 TI - Ensuring local control in meningiomas. PMID- 3042722 TI - On history and heritage: Ralston Paterson. PMID- 3042723 TI - "R.P."--Professor Ralston Paterson. PMID- 3042724 TI - Sandor Ferenczi (1873-1933)--the father of the empathic-interpersonal approach. Part Two: Evolving technique, final contributions and legacy. PMID- 3042725 TI - Bullitt, Freud, and Woodrow Wilson. PMID- 3042726 TI - Psychoanalysis and substance abuse: toward a more effective approach. PMID- 3042727 TI - An economic evaluation of various treatments for contagious foot rot in sheep, using decision analysis. AB - Two decision analysis models were constructed to identify the cost-effectiveness of treatment and/or prevention regimens for ovine foot rot through a systematic evaluation approach of 2 consecutive phases. The first model evaluated the cost effectiveness of examining the sheep for foot lesions and the use of 1 of 32 treatment regimens when lesions were present. The second model evaluated the 6 most cost-effective treatment regimens from the first model with or without a preventive measure (in this case, vaccination) to determine the most cost effective approach to preventing/treating foot rot in a single year. Three prevalence levels of foot rot were used. In addition, 3 levels of reduced production attributable to foot rot were used. Threshold analysis, a form of sensitivity analysis, was used to evaluate allowable variations in the success rates and cost of treatments/prevention where the outcome of the models remains unchanged. Of the 32 treatment regimens tested in this model, the use of 10% zinc sulfate applied to the feet of sheep with foot rot was the most cost-effective regimen, regardless of the prevalence levels of the disease or its impact on productivity. In the second model, the use of vaccination with paring of the feet, as a preventive measure, followed by a treatment of the sheep that were still infected, was less cost-effective than to treat only the sheep with foot lesions and disregard prevention. This was true regardless of the prevalence levels of the disease or its impact on productivity. PMID- 3042728 TI - Severe hypoglycemia attributable to surreptitious injection of insulin in a mare. AB - A mare with signs of hypoglycemia had high serum insulin concentrations before it was euthanatized. High pressure liquid chromatography revealed that the insulin in the mare's blood was of commercial origin. Surreptitious insulin injection has been suspected as the cause of several suspicious deaths of insured horses. The use of high-pressure liquid chromatography should help put an end to this practice. PMID- 3042729 TI - Changes in testicular morphology in boars actively immunized against gonadotropin hormone-releasing hormone. AB - Alterations in testicular morphology were studied in boars actively immunized against gonadotropin hormone releasing hormone (GnRH). Ten boars were divided equally into two experimental groups (five GnRH-immunized, and five controls). Antibody production was achieved by conjugating GnRH to human serum globulin (hSG). The GnRH-hSG conjugate was emulsified in complete Freund's adjuvant, and administered to boars at 12 weeks of age. Boars were given a booster in incomplete Freund's adjuvant on week 18 and 20. The presence of high antibody titers to GnRH caused luteinizing hormone and testosterone to decline to nondetectable levels. Morphometric examination showed a reduction in percentage volume in Leydig cells/unit testis, seminiferous tubule diameter and seminiferous epithelial height, and an increase in non-Leydig cell interstitial tissue in GnRH immunized boars compared with controls. Histologic evaluation displayed severe damage of the seminiferous epithelium, absence of spermatids, incomplete cell associations, disruption of Sertoli cells, formation of multinucleated giant cells, and a striking reduction in size and cytoplasmic structures of Leydig cells in GnRH-immunized animals. These results demonstrate the potent inhibitory effects of GnRH immunoneutralization on the boar reproductive system. PMID- 3042730 TI - Effect of repeated washing on sperm-bound immunoglobulin G. AB - The effect of sperm washing on the stability of sperm-bound immunoglobulin G (IgG) antibody derived from plasma from four patients and also IgG bound in vivo on the spermatozoa of four other men was quantitatively evaluated. In the first series of experiments, human spermatozoa were incubated with an IgG antibody containing plasma and subjected to 18 cycles of sperm washing. In the second set of experiments, spermatozoa from men positive for sperm-bound IgG were subjected to four cycles of sperm washing. The amount of residual antibody bound to a constant number of spermatozoa was quantitated by a radiolabeled antiglobulin assay during and following the washing procedures. There was no significant loss of sperm-bound antibody due to the washing procedures. The results of these studies undermine the utility of sperm washing as an effective treatment of antibody-mediated infertility in men. PMID- 3042731 TI - Structure of a phosphorylated derivative of oleandomycin, obtained by reaction of oleandomycin with an extract of an erythromycin-resistant strain of Escherichia coli. PMID- 3042732 TI - Intravenous administration of ionophores in ruminants: effects on metabolism independent of the rumen. AB - The effect of i.v. administration of ionophores on metabolism in ruminants was investigated in two experiments. In Exp. 1, four Angus heifers were assigned randomly to receive i.v. monensin (18 mg, n = 2) or vehicle (control, n = 2). Samples were collected from indwelling vena cava cannulas from -60 to 240 min. Concentrations of K, Mg (P less than .05) and P (P less than .10) were lower and glucose (GLU) and free fatty acids (FFA) were higher (P less than .05) in monensin-treated than in control heifers. Serum insulin (INS) initially declined and subsequently increased (P less than .05) following monensin administration. A second experiment was conducted to determine the effect of a higher dose of monensin and the effect of lasalocid on minerals and metabolites. Angus (n = 3) and Hereford (n = 3) steers were randomly assigned to treatments in two 3 x 3 latin square designs. Treatments were i.v. administration of monensin, lasalocid or vehicle (ethanol) administered on three consecutive days. Administration of monensin, but not vehicle or lasalocid, resulted in ataxia, hypernea, polyuria and anorexia for approximately 2 h. Plasma concentrations of K, P and Mg were suppressed (P less than .05) by monensin, but not by vehicle or lasalocid administration. The decrease in K was preceded by a transient increase in K 15 min after administering monensin. Concentrations of GLU and FFA increased (P less than .05) following monensin administration. Concentrations of INS were lower from 60 to 120 min and greater at 180 and 240 min compared with -60 to 0 min from monensin administration (P less than .05). These results provide first evidence of an effect of monensin on metabolism in ruminants independent of alterations in ruminal microbial metabolism. PMID- 3042733 TI - Influence of carnitine supplementation on muscle substrate and carnitine metabolism during exercise. AB - We examined 1) the effect of L-carnitine supplementation on free fatty acid (FFA) utilization during exercise and 2) exercise-induced alterations in plasma levels and skeletal muscle exchange of carnitine. Seven moderately trained human male subjects serving as their own controls participated in two bicycle exercise sessions (120 min, 50% of VO2max). The second exercise was preceded by 5 days of oral carnitine supplementation (CS; 5 g daily). Despite a doubling of plasma carnitine levels, with CS, there were no effects on exercise-induced changes in arterial levels and turnover of FFA, the relation between leg FFA inflow and FFA uptake, or the leg exchange of other substrates. Heart rate during exercise after CS decreased 7-8%, but O2 uptake was unchanged. Exercise before CS induced a fall from 33.4 +/- 1.6 to 30.8 +/- 1.0 (SE) mumol/l in free plasma carnitine despite a release (2.5 +/- 0.9 mumol/min) from the leg. Simultaneously, acylated plasma carnitine rose from 5.0 +/- 1.0 to 14.2 +/- 1.4 mumol/l, with no evidence of leg release. Consequently, total plasma carnitine increased. We concluded that in healthy subjects CS does not influence muscle substrate utilization either at rest or during prolonged exercise and that free carnitine released from muscle during exercise is presumably acylated in the liver and released to plasma. PMID- 3042734 TI - Indomethacin, but not dazoxiben, reduced lung fluid filtration after E. coli infusion. AB - Goats were divided into three groups and given infusions of live Escherichia coli bacteria. Group I received no treatment, group II was treated with indomethacin (a cyclooxygenase inhibitor), and group III with dazoxiben (a thromboxane synthase inhibitor). Double indicator-dilution extravascular lung water (EVLW) in group I was significantly different from the treated groups. There was an early increase in EVLW in group I and group III but not in group II animals. At 6 h EVLW's in group I, group II, and group III were 100, 45, and 30% above base line, respectively. Lymph flow (QL) and lymph-to-plasma protein ratio (L/P) was not statistically different between groups. Estimated total fluid filtration [QL + d(EVLW)/dt] in group I and III was markedly elevated between 0 and 1.5-2 h after E. coli infusion. Cardiac output (QT) decreased to 40% of base line in group I, and it decreased slightly in group II because of the indomethacin but did not decrease after E. coli. QT decreased in group III but recovered more rapidly than group I. Mean pulmonary arterial pressure increased more rapidly in group I and reached a higher peak than either treated group. At 6 h these groups had similar pulmonary arterial and pulmonary arterial wedge pressures. We conclude that 1) indomethacin but not dazoxiben blocks the early increase in total fluid filtration after bacterial infusion, 2) dazoxiben does not prevent the increased endothelial permeability resulting from infusion of live bacteria, and 3) indomethacin may somewhat ameliorate the endothelial permeability change.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3042735 TI - Airway reactivity to methacholine in nonatopic asymptomatic adults. AB - We studied 50 nonsmoking volunteers, ages 18-35 yr, with no past or present history or physical examination findings of asthma, rhinitis, allergic disease, or recent respiratory infections, to evaluate the usefulness of the methacholine bronchoprovocation challenge (MBPC) as a screening test for asthma. All were skin test-negative to 29 aeroallergens and had base-line pulmonary function values greater than 80% predicted. Fourteen (28%) subjects had a drop in forced expiratory volume in 1 s (FEV1) of 20% or greater at a provocative dose (PD20FEV1) less than or equal to 225 breath units. Moreover, when these subjects were compared with 21 asymptomatic allergic asthmatics, there was significant overlap between the two groups in concentration of methacholine causing this decline in FEV1. A positive MBPC at methacholine concentrations less than or equal to 5 mg/ml was not diagnostic of asthma, and a negative MBPC at methacholine concentrations greater than or equal to 10 mg/ml did not rule out asthma. These data strongly suggest that MBPC should not be used as the sole factor for the diagnosis of clinically significant asthma. A positive MBPC is one indication of the presence of airway hyperresponsiveness and thus is only one of many factors that must be considered in the diagnosis of asthma. PMID- 3042737 TI - Reflex mechanisms for changes in renal nerve activity during positive end expiratory pressure. AB - This study was designed to investigate the interaction between carotid sinus baroreceptors and cardiopulmonary receptors in the reflex control of renal nerve activity (RNA) during positive end-expiratory pressure (PEEP) in anesthetized dogs. PEEP at two different levels (10 and 20 cmH2O) was applied to the following groups: animals with neuraxis intact (I group, n = 12); vagal and aortic nerve denervated animals with carotid sinus nerves intact (V group, n = 6); carotid sinus denervated animals with vagal and aortic nerves intact (SD group, n = 6); and carotid sinus denervated animals also having severed vagal and aortic nerves (SAV group, n = 12). Mean blood pressure (MBP), central venous pressure, and mean airway pressure were also simultaneously measured. In the I group, no significant alterations in RNA occurred during PEEP at both levels, even when MBP fell significantly. Although the drop in MBP in the SD group was similar to that in the I group, RNA decreased significantly 10 s after intervention at both PEEP levels, followed by a recovery of RNA toward the control level. In contrast, a significant increase in RNA, which continued until the end of PEEP, appeared in the V group immediately after each intervention. In the SAV group, RNA responses to PEEP, which were observed in the other groups, were abolished. These results provide evidence that during PEEP, renal nerve activity is modified by an interaction between carotid sinus baroreceptors and cardiopulmonary receptors; excitatory effects occur via carotid sinus nerves and inhibitory effects occur via vagal afferents. PMID- 3042736 TI - Influenza infection causes airway hyperresponsiveness by decreasing enkephalinase. AB - Ferret tracheal segments were infected with human influenza virus A/Taiwan/86 (H1N1) in vitro. After 4 days, the smooth muscle contractile responses to acetylcholine and to substance P were measured. The response to substance P was markedly accentuated, with a threefold increase in force of contraction at a substance P concentration of 10(-5) M, the highest concentration tested. In contrast, the response to acetylcholine was not affected by viral infection. Histological examination of tissues revealed extensive epithelial desquamation. Activity of enkephalinase (neutral metallo-endopeptidase, EC.3.4.24.11), an enzyme that degrades substance P, was decreased by 50% in infected tissues. Inhibiting enkephalinase activity by pretreating with thiorphan (10(-5) M) increased the response to substance P to the same final level in both infected and control tissues. Inhibiting other substance P-degrading enzymes including kininase II (angiotensin-converting enzyme), serine proteases, and aminopeptidases did not affect the response to substance P. Inhibiting cyclooxygenase and lipoxygenase activity using indomethacin and BW 755c did not affect hyperresponsiveness to substance P. Pretreating tissues with antagonists of alpha-adrenoceptors, beta-adrenoceptors, and H1 histamine receptors (phentolamine 10(-5) M, propranolol 5 X 10(-6) M, and pyrilamine 10(-5) M, respectively) had no effect on substance P-induced contraction. These results demonstrate that infection of ferret airway tissues with influenza virus increases the contractile response of airway smooth muscle to substance P. This effect is caused by decreased enkephalinase activity in infected tissues. PMID- 3042738 TI - Surfactant therapy of newborn rabbits impairs lung macrophage bactericidal activity. AB - Because in vitro studies indicate that pulmonary alveolar macrophages (PAM's) filled with phospholipid vesicles have depressed microbicidal capacity, we tested the intrapulmonary bactericidal activity of newborn PAM's after surfactant treatment. Term newborn rabbits received intratracheally either homologous surfactant or one of two artificial phospholipid vesicle preparations followed by pulmonary aerosol infection with group B streptococci (GBS). Four hours after lung infection, phagocytic killing of GBS was reduced by 70-90% in animals treated with the homologous and one of the artificial surfactants compared with untreated animals or animals that received intrapulmonary injections of the surfactant vehicle (P less than 0.02). The other artificial phospholipid preparation decreased intrapulmonary inactivation of GBS by 30-40% compared with the controls. The phospholipid vesicles in the three preparations were avidly ingested and processed by newborn PAM's. The diminished in vivo killing of GBS was not attributed to decreased viability or phagocytic behavior of the PAM's toward GBS. The bactericidal defect that was evident in the newborn PAM's appeared related to the uptake of large phospholipid vesicles in the preparations rather than to the phospholipid content of the surfactants themselves. When in vitro conditions that stimulated the alveolar environment were used, the natural surfactant preparation promoted GBS proliferation, whereas the artificial preparations did not. Our findings indicate that surfactant administration reduces the bactericidal activity of neonatal PAM's. We conclude that additional investigations are needed to ascertain the effect of surfactant replacement therapy on lost defenses of the lung. PMID- 3042739 TI - Effect of the exercise-induced increase in glucocorticoids on endurance in the rat. AB - To investigate the effect of the increase in glucocorticoids during exercise on endurance, rats were either sham operated (SO) or adrenalectomized. All adrenalectomized rats were given a subcutaneously implanted corticosterone pellet at the time of adrenalectomy. Adrenalectomized rats were injected with corticosterone (ADX Cort) or corn oil (ADX) 5 min before exercise. Rats were killed at rest or after running on a treadmill (21 m/min, 15% grade) until exhaustion. SO rats ran 138 +/- 6 min compared with 114 +/- 9 min for ADX Cort and 89 +/- 8 min for ADX. All differences in run times were significant (P less than 0.05). Corticosterone levels were similar in exhausted SO and ADX Cort groups. ADX exhausted rats had corticosterone levels similar to resting values in SO and ADX rats. Inhibition of the rise in glucocorticoids during exercise had no effect on liver glycogen, liver adenosine 3',5'-cyclic monophosphate, plasma insulin, blood glucose, lactate, glycerol, or 3-hydroxybutyrate, plasma norepinephrine, or red quadriceps and soleus glycogen. Plasma free fatty acids were significantly depressed at exhaustion in ADX rats compared with SO. These data show that glucocorticoids exert effects within the time frame of a prolonged exercise bout and play a role in increasing endurance. PMID- 3042741 TI - Ventilatory failure during loaded breathing: the role of central neural drive. AB - Minute ventilation (VE), arterial blood gases, diaphragmatic electromyogram (EMG) activity, centroid frequency (Fc) and peak inspiratory airway pressures (Paw) were measured in five unanesthetized tracheostomized infant monkeys during various intensities of inspiratory resistive loaded breathing (IRL) until either 1) ventilatory failure occurred (failed trial) or 2) normocapnia was sustained for 1 h (successful trial). During successful trials VE and arterial PCO2 (PaCO2) were sustained at base-line levels, and an increase in peak integrated diaphragmatic EMG activity and peak inspiratory Paw occurred. In contrast, during ventilatory failure runs, VE decreased and PaCO2 rose compared with their respective base-line values. The fall in VE occurred secondary to a significant decline in breathing frequency. Tidal volume was sustained at base-line levels during all trials (both successful and failed groups). Inspiratory Paw's and peak moving time average EMG were sustained at elevated levels during ventilatory failure runs, suggesting that the respiratory muscles did not fail as pressure generators. Furthermore, the EMG Fc did not change from base line during either successful or failed trials. These data suggest that peripheral muscle fatigue did not occur, although in the absence of a more direct test of muscle performance, i.e., a force-frequency curve, we cannot rule out the possibility that a component of peripheral failure contributed to our results. Ventilatory failure during severe IRL in the infant monkey was most clearly associated with an alteration in the respiratory center timing mechanism, i.e., such failure was a function of a decline in respiratory frequency. PMID- 3042740 TI - Relationships among airway reactivity, pupillary alpha-adrenergic and cholinergic responsiveness, and age. AB - Healthy adult volunteers (n = 122), who denied personal history of lung disease or family history of cystic fibrosis or asthma, took no interfering medications, and had forced expiratory volume in 1 s greater than or equal to 80% predicted, underwent methacholine challenge and pupillary reactivity testing. Pupil diameter measured in dark and light test conditions declined with age (Pearson's r = -0.54 and -0.36). Pupillary alpha-adrenergic responsiveness (expressed as the concentration of phenylephrine required to dilate the pupil 1 mm) was significantly correlated with age. Older subjects required lower concentrations for dilation and therefore were more sensitive to phenylephrine. Pupillary cholinergic responsiveness (the concentration of carbachol required for 1-mm constriction) was not significantly correlated with age. Therefore the significantly smaller baseline pupil size in the elderly cannot be explained by failure of alpha-adrenergic receptor responses or by increased pupillary cholinergic responsiveness. We found no significant correlation of methacholine bronchial reactivity with age. In addition, there was no relation between airway reactivity and pupillary alpha-adrenergic or cholinergic responsiveness in this sample of healthy adults. These findings, taken with others in the literature, suggest that the contribution of alpha-adrenergic and cholinergic responsiveness to nonspecific airway reactivity in healthy persons is small, if it exists at all, and that there is no significant change in airway reactivity with age in healthy adults. PMID- 3042742 TI - Involvement of sodium retention hormones during rehydration in humans. AB - We investigated the relation between involuntary dehydration and the mechanisms affecting Na+ retention in the body, focusing on the renin-angiotensin aldosterone system. Six adult males were dehydrated to 2.3% of their body weight by an exercise-heat regimen, followed by rehydration (180 min) with tap water (H2O-R) or 0.45% NaCl solution (Na-R). We measured plasma renin activity (PRA) and aldosterone levels (PA) before dehydration (control), after dehydration, and at 60, 120, and 180 min of rehydration. During the 3-h rehydration period, subjects, restored 51% of the water lost during H2O-R and 71% during Na-R (P less than 0.05). Plasma volume was reduced by an average of 4.5% after dehydration. After 180 min of rehydration, plasma volume restoration during Na-R was to 174% of that lost, and during H2O-R it was to 78% of that lost. We found significant correlations between the change in plasma volume and PRA (r = -0.70, P less than 0.001) and between PRA and PA (r = 0.71, P less than 0.001). In both recovery conditions, PRA increased significantly after dehydration (P less than 0.05) and decreased almost to the control level by 180 min of rehydration, at which time the plasma volume deficit was restored. The change in PA paralleled that in PRA. The rate of sodium excretion was correlated with PA levels in both groups (r = 0.58, P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3042743 TI - Endotoxin causes neutrophil-independent oxidative stress in rats. AB - Endotoxin-induced oxidative stress is investigated in rats by measuring changes in plasma and lung tissue levels of glutathione disulfide (GSSG) using a modified enzymatic assay that allows simultaneous measurement of up to 80 samples. Salmonella enteritidis endotoxin (2 and 20 mg/kg) acutely increased both plasma reduced glutathione and GSSG with a rise in the ratio of GSSG to total glutathione. This increase in GSSG was enhanced by pretreatment with 1,3-bis(2 chloroethyl)1-nitrosourea (BCNU), an inhibitor of the glutathione reductase enzyme. However, there was no significant arteriovenous difference in plasma GSSG across the lung, and lung tissue GSSG did not increase after endotoxin treatment. The increase in plasma GSSG was not blocked by vinblastine-induced neutropenia and could not be reproduced by incubating rat blood in vitro with endotoxin. Receptor antagonists of platelet-activating factor (PAF), at a dose that previously inhibited endotoxin-induced lung injury, attenuated the endotoxin induced increase in plasma GSSG. We conclude that endotoxin causes neutrophil independent oxidative stress in rats, which may be enhanced by the action of platelet-activating factor. PMID- 3042744 TI - Interrupter resistance elucidated by alveolar pressure measurement in open-chest normal dogs. AB - The interrupter method for measuring respiratory system resistance involves rapidly interrupting flow at the mouth while measuring the pressure just distal to the point of interruption. The pressure signal observed invariably exhibits two distinct phases. The first phase is a very rapid jump, designated delta Pinit, which occurs immediately on interruption of flow. The second phase is designated delta Pdif and is a further pressure change in the same direction as delta Pinit but evolving over several seconds. The physiological interpretations of delta Pinit and delta Pdif have been somewhat unclear. Delta Pinit has been taken to equal the pressure drop across the pulmonary airways, possibly with a contribution from the tissues of the respiratory system. Delta Pdif can arise, in principle, from two sources: gas redistribution throughout the lung after interruption of flow and stress recovery within the tissues. To resolve these issues we performed interruption experiments on anesthetized paralyzed, tracheotomized, open-chest normal dogs during passive expiration while measuring alveolar pressures at three sites with alveolar capsules. We found that, in the absence of the chest wall, delta Pinit reflects only the resistance of the airways and that delta Pdif can be ascribed almost entirely to the stress recovery properties of lung tissues. PMID- 3042745 TI - Cutaneous laser-Doppler flowmetry: influence of underlying muscle blood flow. AB - To find whether the measurement of skin blood flow (SkBF) by laser-Doppler flowmetry (LDF) is influenced by blood flow to underlying skeletal muscle, five subjects performed mild forearm exercise to induce a metabolic hyperemia in muscle in both forearms. This exercise consisted of alternative opening and closing of both hands at a frequency of approximately 1/s for a duration of 3 min. This exercise was performed twice by each subject. Forearm blood flow (FBF) by plethysmography increased from 2.64 +/- 0.49 (rest) to 31.11 +/- 9.95 ml.100 ml-1.min-1 (immediately after exercise) (P less than 0.001). No statistically significant postexercise increase was observed in LDF measured on the dorsal (110 +/- 21 to 105 +/- 21 mV) or ventral surface (266 +/- 113 to 246 +/- 77 mV) of the forearm. LDF measured from the chest also showed no significant change, indicating that the exercise was too mild to have reflex effects on SkBF. Moreover, the slope of the logarithmic linear regression and the half-time for recovery during the postexercise period for FBF were not reflected in LDF measurements from any of the three sites. We conclude that LDF measured from the skin surface is not influenced by blood flow to underlying skeletal muscle. PMID- 3042746 TI - The Barthel ADL Index: a standard measure of physical disability? AB - There is no agreed single measure of physical disability for use either clinically or in research. It is argued that acceptance of a single standard measure of activities of daily living (ADL) might increase awareness of disability, improve clinical management of disabled patients, and might even increase acceptance of published research. The Barthel ADL Index is proposed as the standard index for clinical and research purposes. Its validity, reliability, sensitivity, and utility are discussed. The Barthel Index is as good as any other single simple index, and should be adopted as the standard against which future indices are compared. The temptation to use variations on the standard Barthel Index should be resisted. PMID- 3042747 TI - Conditional impairment of cell division and altered lethality in hipA mutants of Escherichia coli K-12. AB - Mutations in hipA, a gene of Escherichia coli K-12, greatly reduce the lethality of selective inhibition of peptidoglycan synthesis. These mutations have also been found to reduce the lethality that accompanies either selective inhibition of DNA synthesis or heat shock of strains defective in htpR. In addition, the mutant alleles of hipA are responsible for a reversible cold-sensitive block in cell division and synthesis of macromolecules, particularly peptidoglycan. Recombination between the chromosome of hipA mutants and plasmids containing noncomplementing fragments of hipA+ revealed that the mutations responsible for both cold sensitivity and reduced lethality were probably identical and, in any case, lay within the first 360 base pairs of the coding region of hipA, probably within the first 50 base pairs. We suggest that the pleiotropic effects of mutations in hipA reflect the involvement of this gene in cell division. PMID- 3042748 TI - Role of uhp genes in expression of the Escherichia coli sugar-phosphate transport system. AB - The uhpABCT locus of Escherichia coli is responsible for expression of the sugar phosphate transport system and its induction by external glucose 6-phosphate. Expression of uhpT-lacZ fusions depended on the function of uhpA, uhpB, and uhpC but not of uhpT. A plasmid carrying only uhpT conferred transport activity in a host strain deleted for the uhp region. Thus, uhpT encodes the polypeptide required for transport function, and the other three uhp genes regulate uhpT transcription. The presence of uhpA at elevated copy number resulted in a substantial increase in uhpT expression. This elevated expression was only about 50% of the level seen in induced haploid cells, and no further increase occurred after addition of inducer. Activation by multicopy uhpA was not affected by the status of uhpC but was decreased in the absence of uhpB, suggesting a role for UhpB in directly activating UhpA. Transcription of uhpA, monitored by expression of a uhpA-lacZ fusion, was not affected by either inducer or the presence of the wild-type uhpA allele. The presence of multiple copies of the uhpT promoter region reduced uhpT expression in strains with uhpA in single copy number but not in those with multiple copies, consistent with competition for the activator. Amino acid sequence comparisons showed that UhpA was homologous to a family of bacterial regulatory proteins, some of which act as transcriptional activators (OmpR, PhoB, NtrC, and DctD). The C-terminal portion of UhpB displayed matches to the corresponding portions of another family of proteins (EnvZ, PhoMR, NtrB, and DctB) that participate in regulation of gene expression in response to environmental factors. PMID- 3042749 TI - Nucleotide sequence and transcription start point of the phosphoglycerate transporter gene of Salmonella typhimurium. AB - We identified the phosphoglycerate transporter gene of Salmonella typhimurium and its polypeptide product and determined the nucleotide sequence of the gene. The predicted translation product was a protein of 406 amino acid residues and was extremely hydrophobic, a feature that is consistent with its role in membrane transport. Hydropathy analysis suggested that there are eight transmembrane segments of at least 20 amino acid residues for the protein. The transcription start point was mapped to lie at position -44 relative to the putative translational initiation start point. Comparison of PgtP with UhpT and GlpT, the membrane-bound proteins involved in the transport of hexose-6-phosphate and glycerol-3-phosphate, respectively, revealed a very high degree of amino acid sequence similarity among them, reflecting not only similar structures and functions among these polypeptides but also a common evolutionary origin for them. PMID- 3042750 TI - Structure and organization of the pel genes from Erwinia chrysanthemi EC16. AB - The pelA and pelC genes from Erwinia chrysanthemi EC16 were sequenced and overexpressed in Escherichia coli cells. These genes and two others from the same strain that were characterized previously encode catalytically related pectate lyase proteins that are involved with the maceration and soft-rotting of plant tissue. The pel genes of strain EC16 were organized as two loosely linked clusters, with two structurally homologous genes in each. The pelA/E cluster also contained the remains of an additional pel gene, the 5' portion of which had been removed by a prior deletion event. Each of the four functional pel genes but not the deleted one contained an efficient rho-independent transcriptional terminator after the translational stop. These and other data indicate that the pel genes are all independently regulated despite their structural homology and tandem clustered organization. Two of the genes, pelA and pelE, encoded proteins that differed greatly in their isoelectric points and ability to macerate plant tissue. A recombinant gene constructed with the 5' portion of pelE and the 3' portion of pelA yielded a chimeric protein with high pectate lyase activity but relatively low maceration activity. This result raised the possibility that the poor maceration ability of the pelA gene product may involve other properties in addition to its low isoelectric point. PMID- 3042753 TI - Mode of peptidoglycan synthesis in Salmonella typhimurium: single-strand insertion. AB - The synthesis of peptidoglycan by Salmonella typhimurium at the molecular level has been analyzed by studying the pattern of insertion of newly synthesized strands into the preexisting cell wall. We have measured the acceptor-donor radioactivity ratio during short labeling periods, and we found values between 0 and 0.2. This is less than the ratio observed by Burman and Park (Proc. Natl. Acad. Sci. USA, 81:1844-1848) for peptidoglycan synthesis in Escherichia coli. We propose that insertion of new strands occurs as single strands. PMID- 3042752 TI - Purification and properties of inosine-guanosine phosphorylase from Escherichia coli K-12. AB - A xanthosine-inducible enzyme, inosine-guanosine phosphorylase, has been partially purified from a strain of Escherichia coli K-12 lacking the deo-encoded purine nucleoside phosphorylase. Inosine-guanosine phosphorylase had a particle weight of 180 kilodaltons and was rapidly inactivated by p chloromercuriphenylsulfonic acid (p-CMB). The enzyme was not protected from inactivation by inosine (Ino), 2'-deoxyinosine (dIno), hypoxanthine (Hyp), Pi, or alpha-D-ribose-1-phosphate (Rib-1-P). Incubating the inactive enzyme with dithiothreitol restored the catalytic activity. Reaction with p-CMB did not affect the particle weight. Inosine-guanosine phosphorylase was more sensitive to thermal inactivation than purine nucleoside phosphorylase. The half-life determined at 45 degrees C between pH 5 and 8 was 5 to 9 min. Phosphate (20 mM) stabilized the enzyme to thermal inactivation, while Ino (1 mM), dIno (1 mM), xanthosine (Xao) (1 mM), Rib-1-P (2 mM), or Hyp (0.05 mM) had no effect. However, Hyp at 1 mM did stabilize the enzyme. In addition, the combination of Pi (20 mM) and Hyp (0.05 mM) stabilized this enzyme to a greater extent than did Pi alone. Apparent activation energies of 11.5 kcal/mol and 7.9 kcal/mol were determined in the phosphorolytic and synthetic direction, respectively. The pH dependence of Ino cleavage or synthesis did not vary between 6 and 8. The substrate specificity, listed in decreasing order of efficiency (V/Km), was: 2' deoxyguanosine, dIno, guanosine, Xao, Ino, 5'-dIno, and 2',3'-dideoxyinosine. Inosine-guanosine phosphorylase differed from the deo operon-encoded purine nucleoside phosphorylase in that neither adenosine, 2'-deoxyadenosine, nor hypoxanthine arabinoside were substrates or potent inhibitors. Moreover, the E. coli inosine-guanosine phosphorylase was antigenically distinct from the purine nucleoside phosphorylase since it did not react with any of 14 monoclonal antisera or a polyvalent antiserum raised against deo-encoded purine nucleoside phosphorylase. PMID- 3042751 TI - In vivo studies of repair of 2-aminopurine in Escherichia coli. AB - The repair of the base analog 2-aminopurine has been studied in vivo by using a temperature-sensitive mutant of the cloned mutH gene of Escherichia coli. Our results suggest that the lethal event in killing of dam mutants by 2-aminopurine does not result simply from incorporation of 2-aminopurine into the DNA and its subsequent repair. Furthermore, a 10-fold increase in the level of 2-aminopurine incorporated into the DNA of a dam mutH double mutant has little effect on the mutation frequency of this strain. An alternative mechanism for the mutagenicity of 2-aminopurine in E. coli is proposed. PMID- 3042754 TI - The ndvA gene product of Rhizobium meliloti is required for beta-(1----2)glucan production and has homology to the ATP-binding export protein HlyB. AB - The ndvA locus of Rhizobium meliloti is homologous to and can substitute for the chvA locus of Agrobacterium tumefaciens. We have previously shown that an ndvA mutant exhibited reduced motility and formed small, white, empty nodules on alfalfa roots. Here we show that this ndvA mutant is defective in the production of the cyclic extracellular polysaccharide beta-(1----2)glucan, even though a 235,000-dalton protein intermediate, known to be involved in the synthesis of this molecule, is present and active in vitro. The DNA sequence of the ndvA locus revealed a single large open reading frame encoding a 67,100-dalton protein that was homologous to a number of bacterial ATP-binding transport proteins. The greatest degree of relatedness was seen with Escherichia coli HlyB, a protein involved in the export of hemolysin, and with the mdr gene product of mammalian cells, which is also homologous to HlyB and thought to be involved in export. Based on the overall symbiotic phenotype of ndvA mutants, the extensive homology between NdvA and HlyB, the fact that ndvA mutants retained an active 235,000 dalton membrane intermediate, and the absence of extracellular beta-(1--- 2)glucan, we propose that NdvA is involved in export of beta-(1----2)glucan from the cell and that this process is fundamentally important for normal alfalfa nodule development. PMID- 3042755 TI - Accumulation of trehalose by Escherichia coli K-12 at high osmotic pressure depends on the presence of amber suppressors. AB - When grown at high osmotic pressure, some strains of Escherichia coli K-12 synthesized substantial levels of free sugar and accumulated proline if it was present in the growth medium. The sugar was identified as trehalose by chemical reactivity, gas-liquid chromatography, and nuclear magnetic resonance spectroscopy. Strains of E. coli K-12 could be divided into two major classes with respect to osmoregulation. Those of class A showed a large increase in trehalose levels with increasing medium osmolarity and also accumulated proline from the medium, whereas those in class B showed no accumulation of trehalose or proline. Most class A strains carried suppressor mutations which arose during their derivation from the wild type, whereas the osmodefective strains of class B were suppressor free. When amber suppressor mutations at the supD, supE, or supF loci were introduced into such sup0 osmodefective strains, they became osmotolerant and gained the ability to accumulate trehalose in response to elevated medium osmolarity. It appears that the original K-12 strain of E. coli carries an amber mutation in a gene affecting osmoregulation. Mutants lacking ADP glucose synthetase (glgC) accumulated trehalose normally, whereas mutants lacking UDP-glucose synthetase (galU) did not make trehalose and grew poorly in medium of high osmolarity. Trehalose synthesis was repressed by exogenous glycine betaine but not by proline. PMID- 3042756 TI - Pausing of flagellar rotation is a component of bacterial motility and chemotaxis. AB - When bacterial cells are tethered to glass by their flagella, many of them spin. On the basis of experiments with tethered cells it has generally been thought that the motor which drives the flagellum is a two-state device, existing in either a counterclockwise or a clockwise state. Here we show that a third state of the motor is that of pausing, the duration and frequency of which are affected by chemotactic stimuli. We have recorded on video tape the rotation of tethered Escherichia coli and Salmonella typhimurium cells and analyzed the recordings frame by frame and in slow motion. Most wild-type cells paused intermittently. The addition of repellents caused an increase in the frequency and duration of the pauses. The addition of attractants sharply reduced the number of pauses. A chemotaxis mutant which lacks a large part of the chemotaxis machinery owing to a deletion of the genes from cheA to cheZ did not pause at all and did not respond to repellents by pausing. A tumbly mutant of S. typhimurium responded to repellents by smooth swimming and to attractants by tumbling. When tethered, these cells exhibited a normal rotational response but an inverse pausing response to chemotactic stimuli: the frequency of pauses decreased in response to repellents and increased in response to attractants. It is suggested that (i) pausing is an integral part of bacterial motility and chemotaxis, (ii) pausing is independent of the direction of flagellar rotation, and (iii) pausing may be one of the causes of tumbling. PMID- 3042757 TI - The product of the F plasmid transfer operon gene, traF, is a periplasmic protein. AB - The products of clones carrying the F plasmid transfer operon gene, traF, were analyzed. Proteins expressed in maxicells were labeled with [35S]methionine and examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. Clones carrying the wild-type traF gene expressed two polypeptide products that were not products of clones containing the traF13 amber mutation. These migrated with apparent molecular weights (Ma) of 27,000 and 25,000. A pulse-chase experiment suggested that the larger product was a precursor of the smaller one. In the presence of ethanol, the Ma-27,000 polypeptide accumulated and the Ma-25,000 product was not expressed. These results indicated that the traF protein undergoes proteolytic processing associated with export. Cell fractionation experiments further indicated that the greatest concentration of the mature (Ma 25,000) TraF protein was located in the periplasm. The DNA sequence of traF and the position of the transition mutation in traF13 DNA were also determined. Sequence analysis suggested that traF would be expressed as a 247-amino-acid, Mr-28,006 polypeptide. The 19 amino acids at the amino terminus of this polypeptide appear to constitute a typical membrane leader peptide, while the remainder of the molecule (Mr 25,942) is predicted to be primarily hydrophilic in character. PMID- 3042759 TI - Physiological regulation of Paracoccus denitrificans methanol dehydrogenase synthesis and activity. AB - An enzyme-linked immunosorbent assay and a whole-cell activity assay were developed which allowed detection of methanol dehydrogenase (MDH) of Paracoccus denitrificans with increased sensitivity. By these methods, it was shown that MDH was not induced by its natural substrate, methanol. Relief from a catabolite repression-like mechanism seemed responsible for low-level MDH synthesis, while product induction was the hypothesized mechanism for synthesis of high amounts of MDH. In the latter process, formaldehyde may play an important role as effector. For a variety of culture conditions, inconsistencies were observed in the relation between amounts of MDH protein synthesized and enzyme activities measured in vitro. Regulation of pyrrolo-quinoline-quinone biosynthesis or a modulation of its incorporation and stability in MDH may constitute an overriding mechanism to ensure a correct tuning between metabolic rates of methanol consumption and the required methanol oxidation rates. PMID- 3042758 TI - Unequal distribution of penicillin-binding proteins among inner membrane vesicles of Escherichia coli. AB - Escherichia coli penicillin-binding proteins (PBPs) were associated only with inner membrane vesicles when separated on 30 to 65% or 19 to 49% (wt/wt) sucrose gradients. Fractionation of vesicles through the low-density gradient revealed at least two classes of PBP-inner membrane associations. The first class consisted of PBPs 1 through 4, and the second class consisted of PBPs 5 through 8. These classes were distinguished by the density of vesicles with which they were associated; class 1 PBPs migrated with vesicles of higher density than did class 2 PBPs. Such combinations suggest that PBPs are nonrandomly distributed within the inner membrane, implying potential functional relationships among the PBPs themselves and with particular membrane domains. In addition, in cell lysates and in vesicle fractions, a 60,000-dalton aztreonam-insensitive PBP or protein fragment was observed which could potentially be confused with PBP3. PMID- 3042760 TI - Penicillin-binding proteins of bdellovibrios. AB - We examined the predacious gram-negative bacterium Bdellovibrio bacteriovorous 109J and free-living strains 109J-A1 and 109J-KA1 derived therefrom for penicillin-binding proteins (PBPs). We compared their PBPs with those of the host bacterium, Escherichia coli, and with those of a facultatively predacious bdellovibrio, B. stolpii UKi2, grown axenically. The multiple PBPs of the 109J strains and of UKi2 differed from each other and from those of E. coli, which suggests that screening for PBPs may be a convenient way to determine to what extent the bdellovibrios may represent a diverse group of organisms. A method for labeling furazlocillin and cefaperizone with iodine-125 is also described. PMID- 3042761 TI - Is efficiency of suppressor tRNAs controlled at the level of ribosomal proofreading in vivo? AB - Ribosomal rpsD mutations did not stimulate nonsense suppressor tRNAs in a general manner according to their increased ribosomal ambiguity and decreased proofreading efficiency. Streptomycin, which stimulates error production by blocking proofreading in vitro, did not increase efficiency of suppressor tRNAs in strains with normal or streptomycin-resistant (rpsL) ribosomes. It did so only in combination with one rpsL mutation which is associated with streptomycin pseudodependence. PMID- 3042762 TI - Germination of Saccharomyces cerevisiae ascospores without trehalose mobilization as revealed by in vivo 13C nuclear magnetic resonance spectroscopy. AB - Saccharomyces cerevisiae ascospores germinate in the presence of acetate without any detectable trehalose degradation, as revealed by high-resolution nuclear magnetic resonance spectroscopy and by a standard colorimetric assay. The results presented here substantiate the hypothesis that in S. cerevisiae trehalose supplies energy during dormancy of the spores and not during the germination process. PMID- 3042763 TI - Direct visualization of RecA protein binding to and unwinding duplex DNA following the D-loop cycle. AB - The RecA protein of Escherichia coli will promote the plectonemic joining of a linear single-stranded DNA molecule with a homologous supertwisted double stranded (ds) DNA molecule. As shown by others, this reaction is characterized by a single cycle of joint formation and dissociation, termed the D-loop cycle. The released DNA products appear by electron microscopy to be topologically identical to the reactant DNAs, yet a second cycle of joining is not observed. This implies that either the RecA protein-single-stranded DNA filament or the dsDNA must be altered during the pairing reaction such that further joint formation is inhibited. Shibata et al. (Shibata, T., DasGupta, C., Cunningham, R. P., Williams, J. K. G., Osber, L., and Radding, C. M. (1982) J. Biol. Chem. 256, 7565 7572) proposed that the dsDNA was inactivated due to the binding of RecA protein following the D-loop cycle, but were unable to describe the structure of the putative RecA-protein-dsDNA complex. Here we have extended those studies to show that if fresh dsDNA is added to a reaction mixture following completion of the D loop cycle, joint formation is stimulated, but only with the freshly added dsDNA. Following completion of the D-loop cycle, a labile RecA protein-dsDNA complex, in which the dsDNA is partially unwound can be preserved by glutaraldehyde fixation and visualized by electron microscopy. This result provides direct evidence that the block to a second cycle of joining is due to the presence of RecA protein remaining bound to the released dsDNA. PMID- 3042764 TI - Lung, heart, and kidney express high levels of mRNA for the vitamin K-dependent matrix Gla protein. Implications for the possible functions of matrix Gla protein and for the tissue distribution of the gamma-carboxylase. AB - We have used cDNA probes for two small vitamin K-dependent bone matrix proteins, bone Gla protein (BGP) and matrix Gla protein (MGP), to evaluate the possibility that either of these proteins might be synthesized by the various soft tissues previously shown to have gamma-carboxylase activity. BGP mRNA was found in bone but not in any of the soft tissues tested, a result which reinforces the view that plasma BGP is a specific marker for bone metabolism. In contrast, MGP mRNA was found in all rat tissues examined. Lung and heart have 10-fold higher levels of MGP mRNA than bone, and kidney has a 5-fold higher level. Despite the high levels of MGP mRNA in heart and kidney, these tissues contain 40-500-fold lower concentrations of MGP protein than bone. Immunofluorescence was used to identify cells that contain MGP in kidney, lung, heart, and spleen. In each tissue, MGP was found in discrete tissue-specific cell types. In most of the soft tissues tested, MGP is the first well characterized substrate for the vitamin K-dependent carboxylase found to be synthesized. The exceptionally broad tissue distribution for MGP synthesis demonstrates that the function of MGP is not specific to connective tissues, and the low levels of MGP antigen in soft tissues with high MGP mRNA levels indicate that MGP is unlikely to act solely by virtue of its accumulation in an extracellular matrix. PMID- 3042765 TI - Escherichia coli transcription termination protein rho has three hydrolytic sites for ATP. AB - We have determined that 3 mol of ATP or other adenine nucleotide can bind to Escherichia coli transcription termination protein rho, in the presence or absence of the RNA cofactor that is required for activation of rho's ATPase activity. Isotope trap experiments show that the three molecules of ATP bound/rho hexamer in the absence of RNA are hydrolyzed upon addition of RNA and are therefore correctly and productively bound at active sites. These results imply that rho acts as a trimer of dimers and that either the ATPase active sites are at the interface between head-to-head protein monomers, or that ATP binding induces asymmetry among rho subunits and results in the formation of functional dimers within the hexamer. We show that ATP is efficiently hydrolyzed by rho only upon RNA binding. We have measured KD values for ATP, ADP, and Pi binding to rho and have constructed a minimal kinetic mechanism for ATP hydrolysis by the enzyme. PMID- 3042766 TI - Enhancement of insulin binding to rat white adipocytes at 15 degrees C by 5,5' dithiobis-(2-nitrobenzoic acid). Independence of the reagent's sulfhydryl group reactivity. AB - Insulin binding to isolated rat white adipocytes at 15 degrees C, a temperature at which cellular degradation of insulin is negligible, has been found to be described by the Two-step Binding Model: R + I in equilibrium RI in equilibrium R'I (Lipkin, E. W., Teller, D. C., and de Haen, C. (1986) J. Biol. Chem. 261, 1702-1711). RI is the initially formed complex between the receptor, R, and insulin, I, and R'I is the complex in an altered state or cellular location. Here the possibility was examined that R'I results from disulfide exchange between the receptor and insulin, an exchange proposed by Clark and Harrison (Clarke, S., and Harrison, L. C. (1986) J. Biol. Chem. 257, 12239-12244) to occur at 37 degrees C. A number of sulfhydryl reagents representing various chemical reactivities did not affect insulin binding. The exception was 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB), which enhanced the number of insulin-binding sites up to 2-fold with no effect on the equilibrium constant. The data suggested that this enhancement was due to activation of cryptic binding sites pre-existing on the cell surface, possibly by increasing the valency of the receptor from 1 to 2. Insulin binding was also enhanced by structural congeners of DTNB devoid of sulfhydryl reactivity, the simplest one being benzoic acid. It was concluded that the effects were not related to modification of sulfhydryl groups, that modification of sulfhydryl groups on the receptor either did not take place or was without effect on binding, and finally, that disulfide exchange between insulin and the receptor was an unlikely explanation for the formation of R'I. Also, since it is possible to show insulin action at 15 degrees C, contrary to the proposal by Clark and Harrison (Clark, S., and Harrison, L. C. (1983) J. Biol. Chem. 258, 11434-11437), disulfide exchange does not appear to be necessary for signal transmission by the occupied receptor. PMID- 3042767 TI - Biochemical characterization of a factor produced by trypomastigotes of Trypanosoma cruzi that accelerates the decay of complement C3 convertases. AB - Infective- and vertebrate-stage trypomastigotes of Trypanosoma cruzi resist serum killing by the alternative complement pathway, whereas noninfective vector-stage epimastigotes, from which trypomastigotes derive, are serum-sensitive. This form of developmental preadaption is commonly observed in protozoan parasites, but its mechanisms are poorly understood. We have demonstrated previously that trypomastigotes spontaneously shed molecules which interfere with formation and accelerate the intrinsic decay of complement C3 convertases, a finding which may explain the evasion of complement lysis by trypomastigotes. We now describe the partial purification and characterization of the T. cruzi C3 convertase inhibitor from the supernatant of culture metacyclic and tissue culture trypomastigotes. Decay-accelerating activity for both classical and alternative pathway C3 convertases copurifies on anion-exchange fast protein liquid chromatography and chromatofocusing with 35S-labeled molecules of 87-93 kDa, pI 5.6-5.8. The labeled components are destroyed by papain and retained on concanavalin A-Sepharose, procedures which remove functional decay-accelerating activity from the supernatant. The 87-93-kDa components are immunoprecipitated by sera from patients chronically infected with T. cruzi, but not by antisera to any known regulatory proteins of the human complement cascade. Lytic activity for tissue culture trypomastigotes in chagasic sera is associated with antibody reactivity against the 87-93-kDa 35S-labeled components and with inhibition of decay accelerating activity. The T. cruzi factor is the first developmentally regulated microbial complement inhibitor to be biochemically characterized. PMID- 3042768 TI - Primary structure of a Plasmodium falciparum malaria antigen located at the merozoite surface and within the parasitophorous vacuole. AB - DNA encoding an antigen of 101,000 apparent molecular weight from the human malaria parasite Plasmodium falciparum was cloned and sequenced. Genomic DNA from the Camp strain covering the complete coding region along with cDNA from the FCR3 strain covering 81% of the coding region were obtained. The cloned DNA specified a full-length protein of 743 amino acids which included two tandemly repeated regions, one near the amino terminus containing eight hexapeptide repeats of sequence TVNDEDED, and the second near the carboxyl terminus containing primarily KE and KEE repeats. The latter repeated region is encoded by a 174-base stretch of mRNA containing only a single pyrimidine. Except for a putative leader sequence located at the amino terminus of the protein, the protein is hydrophilic and highly charged with a calculated isoelectric point of 5.6. Sequences from the Camp and FCR3 strains are very close and are also nearly identical to the partial cDNA sequence of the acidic basic repeated antigen (ABRA) protein from the FC27 strain (Stahl, H.D., Bianco, A.E., Crewther, R.F., Anders, R.F., Kyne, A.P., Coppel, R. L., Mitchell, G.F., Kemp, D.J., and Brown, G.V. (1986) Mol. Biol. Med. 3, 351-368). ABRA was previously shown to be located at the merozoite surface and in the parasitophorous vacuole. Because of its location and because it becomes complexed to merozoites when schizonts rupture in the presence of immune serum, ABRA is a candidate component of a malaria vaccine. PMID- 3042769 TI - Reduced extracellular pH reversibly inhibits oligomerization, intracellular transport, and processing of the influenza hemagglutinin in infected Madin-Darby canine kidney cells. AB - Incubation of Madin-Darby canine kidney cells infected with influenza virus in medium of pH 5.8-6.0 blocks transport of newly synthesized hemagglutinin and processing of the hemagglutinin oligosaccharides to a form resistant to endo H digestion. Upon restoration of the culture medium to pH 7.4, arrested hemagglutinin is processed and then appears on the cell surface, indicating that exclusively transport and not oligosaccharide processing is inhibited. Based upon kinetic data and localization of blocked hemagglutinin by immunofluorescence, the point of inhibition appears to be a discrete step in transport located in a pre Golgi compartment. This conclusion is supported by the observation that trimerization of hemagglutinin, which is believed to occur in the endoplasmic reticulum, is also inhibited by acidic medium. PMID- 3042770 TI - Sequence and domain structure of yeast pyruvate carboxylase. AB - The nucleotide sequence of the yeast pyruvate carboxylase gene has been determined from a cloned fragment of yeast genomic DNA. The deduced translation product codes for a polypeptide of 1178 amino acids, having a calculated molecular weight of 130,100. The protein shows strong sequence homology to specific regions of other biotin carboxylases, lipoamide transferases, and carbamyl phosphate synthetases. The homologous regions suggest the presence of three subsites in the enzyme: a biotin attachment site, a keto acid-binding site, and an ATP-binding site. Partial proteolysis with a variety of proteases under nondenaturing conditions indicates the presence of structural domains corresponding to these subsites. PMID- 3042771 TI - Structural characterization of Escherichia coli phosphatidylserine decarboxylase. AB - Phosphatidylserine decarboxylase of Escherichia coli is one of a small group of pyruvoyl-dependent enzymes (Satre, M., and Kennedy, E.P. (1978) J. Biol. Chem. 253, 479-483). The DNA sequence of the structural gene (psd) and partial protein sequence studies demonstrate that the enzyme contains two nonidentical subunits, alpha (Mr = 7,332) and beta (Mr = 28,579), which are derived from a single proenzyme. These two subunits are blocked at their respective amino termini. Reduction of the enzyme with NaCNBH3 in the presence of radiolabeled phosphatidylserine resulted in association of the label with the alpha subunit. Similar reduction in the presence of ammonium ions exposed a new amino terminus for the alpha subunit beginning with alanine. Therefore, the pyruvate prosthetic group is in amide linkage to the amino terminus of the alpha subunit. The amino terminus of the beta subunit was determined to be formylmethionine. The carboxyl terminus of the beta subunit was determined to be glycine as predicted by the DNA sequence. Comparison of the DNA sequence and protein sequence information revealed that the decarboxylase is made as a proenzyme (Mr = 35,893), and the predicted amino acid at the position of the pyruvate within the open reading frame of the proenzyme is serine. Therefore, as with other pyruvoyl-dependent decarboxylases, the prosthetic group is derived from serine through a post translational cleavage of a proenzyme. PMID- 3042772 TI - Effects of Escherichia coli secB mutations on pre-maltose binding protein conformation and export kinetics. AB - Mutations affecting the secB gene of Escherichia coli cause a defect in protein export. This report presents the demonstration that the secB mutations caused a defect in co-translational processing of maltose binding protein (MBP). A significant amount of post-translational processing of pre-MBP occurred within 1 min after termination of pulse labeling; at later time points only a small amount of additional processing occurred. The conformation of the intracellular precursor form of MBP was examined in a secB::Tn5 mutant, using protease sensitivity (Randall, L. L., and Hardy, S. J. S. (1986) Cell 46, 921-928) as the assay. In contrast to the isogenic wild type strain, a population of pre-MBP that had folded into a protease resistant conformation was detected in the secB mutant. In addition, sublethal doses of chloramphenicol did not significantly affect protein export in the secB::Tn5 mutant and the secB::Tn5 mutation did not lead to defects in membrane energization. PMID- 3042773 TI - Chemical composition of the yeast ascospore wall. The second outer layer consists of chitosan. AB - In a preceding paper (Briza, P., Winkler, G., Kalchhauser, H., and Breitenbach, M. (1986) J. Biol. Chem. 261, 4288-4294), we reported the presence of dityrosine in the outer layers of yeast ascospore walls. Both outer layers seen in electron micrographs of yeast ascospore walls are sporulation-specific. Here we show that the second of these two outer layers consists of chitosan. In intact spores, it is shielded from staining with primulin by the outermost layer. However, in purified spore walls, the second layer is brightly stained by primulin, and hydrolysates of such preparations contain about 10% glucosamine relative to spore wall dry weight. The spore wall material staining with primulin is resistant to chitinase, but readily degraded by treatment with HNO2. Acetylation prior to HNO2 treatment completely prevents its degradation. A partial acid hydrolysate of spore walls contains predominantly soluble poly-beta-(1,4)-glucosamine as determined by 13C NMR spectroscopy. By these criteria, the glucosamine polymer of yeast ascospore walls is chitosan. As spore walls treated with alkali lack the inner layers but contain chitosan and as chitosan is not exposed at the surface of the spore, we conclude that it is localized in the second outer layer of the spore wall. PMID- 3042774 TI - Inhibitor-induced enzyme activation in organic solvents. AB - The enzymatic activity of the protease subtilisin in anhydrous organic solvents can be dramatically increased by pretreating the enzyme before it is placed in the nonaqueous medium. For instance, lyophilization of subtilisin from aqueous solution containing competitive inhibitors (followed by their removal) created an enzyme which was up to 100 times more active than the enzyme lyophilized in the absence of such ligands. This phenomenon of ligand-induced "enzyme memory" also extends to the stability, affinity, and substrate specificity of subtilisin in organic solvents. PMID- 3042775 TI - Role of cysteine residues in glutathione synthetase from Escherichia coli B. Chemical modification and oligonucleotide site-directed mutagenesis. AB - Escherichia coli B glutathione synthetase is composed of four identical subunits; each subunit contains 4 cysteine residues (Cys-122, -195, -222, and -289). We constructed seven different mutant enzymes containing 3, 2, or no cysteine residues/subunit by replacement of cysteine codons with those of alanine in the gsh II gene using site-directed mutagenesis. Three mutant enzymes, Ala289, Ala222/289, Cys-free (Ala122/195/222/289), in which cysteine at residue 289 was replaced with alanine, were not inactivated by 5,5'-dithiobis(2-nitrobenzoate) (DTNB), while the other four mutants retaining Cys-289 were inactivated at the wild-type rate. From these selective inactivations of mutant enzymes by DTNB, the sulfhydryl group modified by DTNB was unambiguously identified as Cys-289. In this way, Cys-289 was found to be also a target of modification with 2 nitrothiocyanobenzoate and N-ethylmaleimide, while Cys-195 was of p chloromercuribenzoate. These results suggest that both Cys-195 and Cys-289 were not essential for the activity of the glutathione synthetase, but chemical modification of either one of the two sulfhydryl groups resulted in complete loss of the activity. Replacement of Cys-122 to Ala-122 enhanced the reactivity of Cys 289 with sulfhydryl reagents. PMID- 3042776 TI - Purification and properties of ferrochelatase from the yeast Saccharomyces cerevisiae. Evidence for a precursor form of the protein. AB - Ferrochelatase was purified to homogeneity from yeast mitochondrial membranes and found to be a 40-kDa polypeptide with a pI at 6.3. Fatty acids were absolutely necessary to measure the activity in vitro. The Michaelis constants for protoporphyrin IX (9 x 10(-8) M), ferrous iron (1.6 x 10(-7) M), and zinc (9 x 10(-6) M) were determined on purified enzyme preparations in the presence of dithiothreitol. However, the Km for zinc was lower when measured in the absence of dithiothreitol (K-m(Zn2+) = 2.5 x 10(-7) M, Km(protoporphyrin) unchanged). The maximum velocities of the enzyme were 35,000 nmol of heme/h/mg of protein and 27,000 nmol of zinc-protoporphyrin/h/mg of protein. Antibodies against yeast ferrochelatase were raised in rabbits and used in studies on the biogenesis of the enzyme. Ferrochelatase is synthesized as a higher molecular weight precursor (Mr = 44,000) that is very rapidly matured in vivo to the Mr = 40,000 membrane bound form. This precursor form of ferrochelatase was immunoprecipitated from in vitro translation (in a rabbit reticulocyte lysate system) of total yeast RNAs. The antibodies were used to characterize two yeast mutant strains deficient in ferrochelatase activity as being devoid of immunodetectable protein in vivo and ferrochelatase mRNA in vitro translation product. The N-terminal amino acid sequence of the purified protein has been established and was found to be frayed. PMID- 3042777 TI - Biosynthesis of diphthamide in Saccharomyces cerevisiae. Partial purification and characterization of a specific S-adenosylmethionine:elongation factor 2 methyltransferase. AB - The inactivation of elongation factor 2 (EF-2) by diphtheria toxin requires the presence of a post-translationally modified histidine residue in EF-2. This residue, diphthamide, has the structure 2-[3-carboxyamido-3 (trimethylammonio)propyl]histidine. The present work was undertaken to study the pathway of diphthamide biosynthesis using diphtheria toxin-resistant yeast mutants (Chen. J.-Y., Bodley, J. W., and Livingston, D. M. (1985) Mol. Cell. Biol. 5, 3357-3360) which are defective in diphthamide formation. We demonstrate here that one of these mutants (dph5) contains a toxin-resistant form of EF-2 which can be converted in vitro to a toxin-sensitive form through the action of an enzyme present in other yeast strains. Both this toxin-resistant EF-2 and its modifying enzyme have been partially purified and evidence is presented that the modifying enzyme is a specific S-adenosylmethionine:EF-2 methyltransferase. In vitro complementation to diphtheria toxin sensitivity required S adenosylmethionine, and when partially purified components were incubated with [methyl-3H]S-adenosylmethionine, label was incorporated specifically into EF-2. Hydrolysis of labeled EF-2 yielded diphthine (the unamidated form of diphthamide) and a single chromatographically separable labeling intermediate. We conclude that the S-adenosylmethionine:EF-2 methyltransferase adds at least the last two of the three methyl groups present in diphthine and that this modification is sufficient to create diphtheria toxin sensitivity. Evidence is also presented for the existence of an ATP-dependent amidating enzyme which catalyzes the final step in the biosynthesis of diphthamide in EF-2. PMID- 3042778 TI - SEC7 encodes an unusual, high molecular weight protein required for membrane traffic from the yeast Golgi apparatus. AB - Saccharomyces cerevisiae with mutations at the sec7 locus are pleiotropically deficient in protein transport within the Golgi apparatus and proliferate a large array of Golgi cisternae at a restrictive growth temperature (37 degrees C). The SEC7 gene and its product (Sec7p) have been evaluated by molecular cloning and sequence analysis. Two genes that allow sec7 mutant cells to grow at 37 degrees C are represented in wild-type yeast DNA libraries. A single copy of the authentic SEC7 gene permits growth of mutant cells, whereas the other gene suppresses growth deficiency only when expressed from a multicopy plasmid. The SEC7 gene is contained on a 8.4-kilobase pair SphI restriction fragment, portions of which hybridize to a single 6-kilobase pair mRNA. The gene is essential for yeast vegetative growth. DNA sequence analysis of this region detects a single open reading frame with the potential to encode a 2008-amino acid-long hydrophilic protein of 230 kDa. Putative Sec7p contains an unusual, highly charged acidic domain of 125 amino acids with 29% glutamate, 18% aspartate, and 21% serine. Within this region, stretches of 14 consecutive glutamate residues and 13 consecutive glutamates/aspartates are predicted. This domain in Sec7p may serve a structural role to interact with lipids or proteins on the cytoplasmic surface of the Golgi apparatus. PMID- 3042780 TI - Biochemical properties of Ha-ras encoded p21 mutants and mechanism of the autophosphorylation reaction. AB - Kinetic studies performed on p21H guanine nucleotide complexes with and without Mg2+ show that point mutations at positions 12, 59, and 61 each have a different effect on the rate of nucleotide dissociation. Double mutants with a combination of these amino acid substitutions reveal that the effects of each mutation on these kinetics are interactive (nonadditive) for positions 12 and 59 and approximately additive for the positions 12 and 61. The magnitude and direction of the effects seen are dependent on the nature of the nucleotide and whether or not the complexes contain Mg2+. All the mutants have reduced GTPase activity. It is also shown that the autophosphorylation reaction velocity is of first order with respect to the protein concentration and that this reaction is an intramolecular one, which takes place as a side reaction of the GTPase reaction. The autophosphorylation is not reversible under the experimental conditions. The covalently bound phosphate does not decrease the nucleotide-binding ability of the protein nor does it change the relative affinity of the protein for GTP versus GDP. The results are discussed in terms of the structural model and function of p21H. PMID- 3042779 TI - Sequence of the lon gene in Escherichia coli. A heat-shock gene which encodes the ATP-dependent protease La. AB - To learn more about the mechanism and regulation of the ATP-dependent protease La in Escherichia coli, the lon gene was completely sequenced using the dideoxy method on fragments generated by Bal31 digestion. The predicted amino acid composition based on the DNA sequence agreed well with the composition of the acid-hydrolyzed protease. The predicted NH2-terminal amino acid sequence, tryptophan content, and the carboxyl terminus also agreed with experimental data. However, the molecular weight of 87,000 (783 amino acids) calculated from the DNA sequence was lower than prior estimates. The tetrameric enzyme contains four binding sites for ATP, a DNA-binding domain, a proteolytic site, and a regulatory site that binds unfolded polypeptides. An ATP-binding pocket exists on each subunit as shown by consensus sequences and elements of secondary structure resembling those on other nucleotide-binding proteins (e.g. adenylate kinase, RecA). For this purpose, improved consensus patterns for identifying ATP-binding domains were developed. Computer-assisted comparisons, however, failed to demonstrate any regions homologous to sequences in other polypeptides including proteases or DNA-binding proteins. This enzyme also contains an unusual highly acidic domain surrounded by very basic sequences. Protease La is the first ATP dependent protease sequenced and seems to represent a new type of enzyme. The promoter sequence was similar to consensus sequences for other heat-shock promoters. Using site-directed mutagenesis, alterations were introduced into the putative promoter sequence. Mutations upstream of -35 had little effect, but alterations immediately upstream of -10 lowered basal transcription of a lon-lacZ operon fusion and reduced its response to inducers of the heat-shock response. PMID- 3042781 TI - Analogs of diaminopimelic acid as inhibitors of meso-diaminopimelate dehydrogenase and LL-diaminopimelate epimerase. AB - Analogs 1-8 of diaminopimelic acid (DAP) were synthesized and tested for inhibition of purified meso-DAP D-dehydrogenase from Bacillus sphaericus and of LL-DAP epimerase from Escherichia coli. The dehydrogenase was assayed by monitoring NADPH formation spectrophotometrically at 340 nm. N-Hydroxy DAP 4, N amino DAP 5, and 4-methylene DAP 6 are substrates of the dehydrogenase with relative rates exceeding those of the meso isomers of the thia analogs 1ab, 2ab, and 3ab. DAP epimerase was assayed by coupling the epimerization of LL-DAP to DL DAP (Km = 0.26 mM) with the dehydrogenase-catalyzed oxidation of DL-DAP by NADP. Lanthionine isomers 1ab and 1c were stronger inhibitors of the epimerase (Ki = 0.18 mM, Ki' = 0.67 mM, and Ki = 0.42 mM, respectively) than the corresponding meso-sulfoxide 2ab or the meso-sulfone 3ab. Other isomers of 2 and 3, as well as compounds 7 and 8, showed no epimerase inhibition. N-Hydroxy DAP 4 was the most potent competitive inhibitor (Ki = 0.0056 mM) of the epimerase, whereas N-amino DAP 5 is weaker (Ki = 2.9 mM) and 4-methylene DAP 6 is a noncompetitive inhibitor (Ki' = 0.95 mM). Although none of the analogs tested showed time-dependent inactivation of either enzyme, compounds 4, 5, 6, and 7 display substantial antibacterial activities. Possible mechanisms of epimerase inhibition and significance of the DAP pathway as a target for antibiotics are discussed. PMID- 3042783 TI - Inhibition of sterol synthesis by delta 5-sterols in a sterol auxotroph of yeast defective in oxidosqualene cyclase and cytochrome P-450. AB - Synthesis of ergosterol is demonstrated in the GL7 mutant of Saccharomyces cerevisiae. This sterol auxotroph has been thought to lack the ability to synthesize sterols due both to the absence of 2,3-oxidosqualene cyclase and to a heme deficiency eliminating cytochrome P-450 which is required in demethylation at C-14. However, when the medium sterol was 5 alpha-cholestan-3 beta-ol, 5 alpha cholest-8(14)-en-3 beta-ol, or 24 beta-methyl-5 alpha-cholest-8(14)-en-3 beta-ol, sterol synthesis was found to proceed yielding 1-3 fg/cell of ergosterol (24 beta methylcholesta-5,7,22E-trien-3 beta-ol). Ergosterol was identified by mass spectroscopy, gas and high performance liquid chromatography, ultraviolet spectroscopy, and radioactive labeling from [3H]acetate. Except for some cholest 5-en-3 beta-ol (cholesterol) which was derived from the 5 alpha-cholestan-3 beta ol, the stanol and the two 8(14)-stenols were not significantly metabolized confirming the absence of an isomerase for migration of the double bond from C 8(14) to C-7. Drastic reduction of ergosterol synthesis to not more than 0.06 fg/cell was observed when the medium sterol either had a double bond at C-5, as in the case of cholesterol, or could be metabolized to a sterol with such a bond. Thus, both 5 alpha-cholest-8(9)-en-3 beta-ol and 5 alpha-cholest-7-en-3 beta-ol (lathosterol) were converted to cholesta-5,7-dien-3 beta-ol (7 dehydrocholesterol), and the presence of the latter dienol depressed the level of ergosterol. The most attractive of the possible explanations for our observations is the assumption of two genetic compartments for synthesis of sterols, one of which has and one of which has not been affected by the two mutations. The ability, despite the mutations, to synthesize small amounts of ergosterol which could act to regulate the cell cycle may also explain why this mutant can grow aerobically with cholesterol (acting in the bulk membrane role) as the sole exogenous sterol. PMID- 3042782 TI - Interaction of a folded chromosome-associated protein with single-stranded DNA binding protein of Escherichia coli, identified by affinity chromatography. AB - A single-stranded DNA-binding protein (SSB) affinity column was prepared by optimizing the coupling of Escherichia coli single-stranded DNA-binding protein to Affi-Gel 10. The bound SSB retained its ability to specifically bind single stranded DNA. When nuclease-treated cell extracts were incubated with the SSB beads overnight at 4 degrees C, a major protein of Mr = 25,000 was bound. At shorter incubation times, two additional proteins of Mr = 32,000 and 36,000 were also detected. In the absence of nuclease treatment, eight additional proteins ranging from Mr = 14,000 to 160,000 also bound to the affinity column. The major Mr = 25,000 protein has been shown to be a folded chromosome-associated protein. Its binding to SSB is strongly enhanced by the addition of DNA polymerase III or DNA polymerase III holoenzyme. PMID- 3042784 TI - Ca2+ and pH responses to sequential additions of mitogens in single 3T3 fibroblasts: correlations with DNA synthesis. AB - The progression of Swiss 3T3 fibroblasts from the quiescent state (G0) through G1 to DNA synthesis in S phase generally requires the synergistic action of two mitogens. The main aim of this study was to compare systematically the early Ca2+ and pH responses in quiescent cells to all of the pair combinations of eight mitogens (bombesin, platelet-derived growth factor, vasopressin, prostaglandin F2 alpha, epidermal growth factor, 12-O-tetradecanoyl phorbol-13-acetate, insulin, 8 bromo-cAMP) with their subsequent effects on DNA synthesis. Each of the mitogens which caused inositol phosphate accumulation (bombesin, platelet-derived growth factor, vasopressin, prostaglandin F2 alpha) also activated Ca2+- and phospholipid-dependent protein kinase (protein kinase C) and generated both the Ca2+ and pH responses, although epidermal growth factor also generated the ionic responses without causing release of inositol phosphates or activation of protein kinase C. For sequential mitogen additions the ionic signals were measured in single cells as well as in cell populations to avoid ambiguities due to heterogeneity in the responses of the cells to the various mitogens. The modulating effects of the mitogens on the [Ca2+]i responses to subsequent mitogen additions varied widely, but detailed comparisons showed that the pattern of blocking effects could not be attributed solely to the effect of the first mitogen causing either maximal breakdown of phosphatidylinositol 4,5-bisphosphate or complete depletion of the intracellular Ca2+ pool or activation of protein kinase C. From these analyses it was concluded that the requirement for two mitogens for effective DNA synthesis could not be attributed to the summation to a critical threshold of either the ionic signals or phosphatidylinositol 4,5 bisphosphate breakdown, and that these responses are insufficient by themselves to cause the cells to progress to DNA synthesis in S phase. PMID- 3042785 TI - Novel feature of metabolism of low density lipoprotein receptor in a mouse macrophage-like cell line, J774.1. AB - Biosynthesis, processing, and degradation of low density lipoprotein (LDL) receptors were studied in a mouse macrophage-like cell line, J774.1, by immunoprecipitation and immunoblotting with an antibody directed against the COOH terminal 14 amino acids of the LDL receptor. The molecular weight of the mature LDL receptor of J774.1 cells maintained in RPMI medium was 140,000 under nonreducing condition and 160,000 under reducing condition in sodium dodecyl sulfate-polyacrylamide gels. These sizes are 10,000-15,000 daltons larger than those of the receptor in other mouse fibroblastic cells or P388 leucocyte. However, when J774.1 cells were cultured in Dulbecco's modified Eagle's medium, the molecular weight of the mouse cell lines, 123,000 under nonreducing condition and 153,000 under reducing condition. The larger LDL receptor molecules produced by J774.1 cells cultured in RPMI were insensitive to the treatment with end-alpha N-acetylgalactosaminidase (O-glycanase), suggesting that aberrant serine/threonine-linked (O-linked) glycosylation might account for the apparent large size. Pulse-chase experiments revealed that the rate of processing of the LDL receptor from precursor to mature form in J774.1 was similar to that in other mouse cell lines, but the rate of degradation was much faster: half-life of the LDL receptor of J774.1 was about 2 h. No significant difference in biological function or lifetime was observed between the normal and the larger LDL receptor. This novel character of molecular size and lifetime of the LDL receptor in J774.1 is discussed in relation to altered maturation and/or modification during receptor biosynthesis. PMID- 3042786 TI - Asparaginase II of Saccharomyces cerevisiae. Characterization of the ASP3 gene. AB - Purified preparations of asparaginase II of Saccharomyces cerevisiae exhibit two protein bands upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Cloning and sequencing of the ASP3 gene, and partial amino acid sequencing as asparaginase II, imply that both bands are encoded by ASP3 but have different N termini. Northern blot analysis using the cloned ASP3 gene as a probe indicates that nitrogen catabolite repression of asparaginase II is achieved by alteration in mRNA levels. Deletion of sequences greater than 600 base pairs upstream from the initiation AUG codon results in an altered response to certain nitrogen sources in strains containing the truncated gene. PMID- 3042787 TI - Nucleotide sequence of the human placental alkaline phosphatase gene. Evolution of the 5' flanking region by deletion/substitution. AB - Three closely related alkaline phosphatase (ALP) genes reside on the long arm of chromosome 2 in man. One of these genes (the placental ALP-1) encodes the classic heat-stable placental alkaline phosphatase. Another gene (the placental ALP-2) is closely related to the placental ALP-1 and may encode the so-called placental ALP like enzyme of the testis and thymus. The third member of this gene family (the intestinal ALP gene) encodes the intestinal alkaline phosphatase. The expression of the placental ALP-1 and intestinal ALP genes is highly tissue-specific in spite of nearly 90% sequence similarity within their exons. To help determine the basis for this tissue specificity, the nucleotide sequence of the placental ALP-1 gene and some of its 5' flanking region has been determined and analyzed by comparison with placental ALP-2 and intestinal ALP gene sequences. The placental ALP-1 gene transcription unit has 4087 bases between the major cap site and the most distal of several reported 3' ends. The protein coding region is divided by 10 short introns varying in size from 74 to 241 nucleotides. Three of these introns bisect regions of the gene that encode residues conserved between the active site of the Escherichia coli enzyme and the human placental ALP. This result suggests that the human alkaline phosphatase genes have evolved in an intron-independent fashion. A comparison of the placental ALP-1 5' flanking sequence (up to -540) with the analogous sequence of the intestinal ALP gene revealed several deletion/substitutions which could be important in determining the tissue-specific expression of these genes. PMID- 3042788 TI - Ketopantoic acid reductase of Pseudomonas maltophilia 845. Purification, characterization, and role in pantothenate biosynthesis. AB - Ketopantoic acid reductase (EC 1.1.1.169), an enzyme that catalyzes the formation of D-(-)-pantoic acid from ketopantoic acid, was purified 6,000-fold to apparent homogeneity with a 35% overall recovery from Pseudomonas maltophilia 845 and then crystallized. The relative molecular mass of the native enzyme, as estimated by the sedimentation equilibrium method, is 87,000 +/- 5,000, and the subunit molecular mass is 30,500. The enzyme shows high specificity for ketopantoic acid as a substrate (Km = 400 microM, Vm = 1,310 units/mg of protein) and NADPH as a coenzyme (Km = 31.8 microM). Only 2-keto-3-hydroxyisovalerate (Km = 8.55 mM, Vm = 35.8 units/mg) was reduced among a variety of other carbonyl compounds tested. The reaction is reversible (Km for D-(-)-pantoic acid = 52.1 mM), although the reaction equilibrium greatly favors the direction of D-(-)-pantoic acid formation. That the enzyme is responsible for the synthesis of D-(-)-pantoic acid necessary for the biosynthesis of pantothenic acid in P. maltophilia 845 is indicated by the observations that only this enzyme is missing in D-(-)-pantoate (or pantothenate)-requiring mutants derived from P. maltophilia 845 among several enzymes (i.e. ketopantoyl lactone reductase (EC 1.1.1.168) and acetohydroxy acid isomeroreductase (EC 1.1.1.86], which may be concerned in the formation of D-(-) pantoic acid, assayed, whereas it is present in substantial amounts in the parent strain and in spontaneous revertants of the mutants. PMID- 3042789 TI - Import of the malate dehydrogenase precursor by mitochondria. Cleavage within leader peptide by matrix protease leads to formation of intermediate-sized form. AB - The mitochondrial matrix enzyme malate dehydrogenase (MDH) is synthesized on cytoplasmic polysomes as a larger precursor (pMDH) with an NH2-terminal leader peptide of 24 amino acids. Import of in vitro synthesized MDH into mitochondria results in formation of the mature-sized subunit. We report here that the conversion of pMDH to mMDH occurs via two distinct cleavage events within the leader peptide. First, pMDH is cleaved to an intermediate form (iMDH) of MDH. Conversion of the precursor to the intermediate form is catalyzed by a protease localized to the mitochondrial matrix. The cleavage of pMDH to iMDH involves the removal of 15 amino acids from the NH2 terminus of the pMDH leader peptide. The iMDH is subsequently cleaved, also by a matrix protease, to mature MDH in a reaction which is O-phenanthroline-sensitive. Cleavage to iMDH and to mature MDH occurs prior to completion of translocation of the MDH polypeptide chain into the mitochondrial matrix. PMID- 3042790 TI - Expression of phosphoprotein p19 in brain, testis, and neuroendocrine tumor cells. Developmental regulation in rat brain. AB - We have recently purified from bovine brain a 19-kDa protein, p19, that was previously shown to undergo hormonally regulated phosphorylation in several neuroendocrine tumor cells. We now report the tissue distribution of p19, studied by immunoblotting. Using a rabbit antiserum, which binds both to the unphosphorylated form and to the two predominant phosphoforms of p19, we show that the protein is present in brain and testis but not in a variety of other mammalian tissues. High levels of p19 are also present in several cultured tumor cells expressing neuroendocrine properties. In addition, p19 was detected in HL60 promyelocytic leukemia and in Friend erythroleukemia cells, but not in several other cell lines. In rat brain, we show that the level of p19 is maximal on the first postnatal day and declines within the first 2 weeks of life to a low plateau that persists into adulthood. The concentration of translatable p19 mRNA also decreases postnatally in rat brain, suggesting that the developmental regulation of the expression of p19 occurs, at least in part, at a pretranslational level. The broad species cross-reactivity of the p19 antibody suggests that the gene encoding p19 has been highly conserved during mammalian evolution. Based on the pattern of expression of this protein, we propose that p19 plays a role in the development of neurons and neuroendocrine cell types. PMID- 3042791 TI - Cortical bone repair. The relationship of the lacunar-canalicular system and intercellular gap junctions to the repair process. AB - Repair of cortical bone was studied in 2.4-millimeter-diameter mid-diaphyseal femoral and tibial defects in young New Zealand White rabbits using light microscopy, transmission electron microscopy, and histomorphometry. The initial source of repair tissue is the marrow. Vessels grow into the defect, accompanied by undifferentiated mesenchymal cells. Woven bone is synthesized initially at the periphery of the defect on pre-existing cortex. Differentiating mesenchymal osteoblasts surround themselves with osteoid in a woven conformation. Once a scaffold has formed, surface osteoblasts align themselves in a regular array on the woven matrix surface and synthesize osteoid in a lamellar conformation. The long axes of the repair vessels, lamellae, and osteocyte lacunae are perpendicular to the long axis of the bone. Polarized-light microscopy showed maintenance of this pattern at six, eight, and twelve weeks, even when the defect was filled with lamellar bone. Remodeling is performed slowly by osteoclast cutting cones over a period of several months. The lacunar-canalicular system is clearly demonstrated in plastic-embedded, toluidine blue-stained sections. A canaliculus passes into or away from a lacuna every 1.9 micrometers over the entire osteocyte perimeter. Undifferentiated mesenchymal cells have no processes, as seen by transmission electron microscopy, but soon sprout a florid array of processes as differentiation to early mesenchymal osteoblasts proceeds. Osteoblast and osteocyte cell processes are packed with intermediate filaments that are continuous with those in the cell bodies. Intercellular gap junctions are seen between surface osteoblasts, between osteoblasts and underlying osteocytes, and between osteocyte cell processes in the canaliculi. PMID- 3042792 TI - Operative treatment of congenital idiopathic club foot. PMID- 3042793 TI - Treatment of ununited fractures of the scaphoid by iliac bone grafts and Kirschner-wire fixation. AB - Of 151 ununited fractures of the scaphoid that were treated with iliac bone grafts and Kirschner-wire fixation through a volar approach, all but four (97 per cent) healed in an average of seventeen weeks, Three of the four failures resulted from obvious technical errors. Neither the preoperative existence of necrosis of the proximal fragment nor the location of the fracture affected the results. When there was mild radiocarpal arthritis preoperatively, it did not progress postoperatively; if there was moderate radiocarpal arthritis preoperatively, progression seldom was seen if a radial styloidectomy was done. Displaced and unstable ununited fractures healed even if the deformity was not corrected completely. The principal benefit of the procedure was relief of pain rather than an increase either in motion of the wrist or in strength of grip. PMID- 3042794 TI - Stab wounds involving the brachial plexus. A review of operated cases. AB - Of 64 patients with stab wounds involving the brachial plexus operated on by one surgeon, 18 were followed up in detail, with a view to reviewing operative techniques, results and the lessons to be learned. Primary nerve grafting produced better results than end-to-end repair, even in fresh cases. The recognition of nerve compression by a false aneurysm is important, since in these cases, vascular repair alone may not give recovery and neurolysis may be necessary. The overall results of operation were good; lesions of C5 and C6 roots recovered better than those of more distal roots. PMID- 3042795 TI - Honour volume on the occasion of the 80th birthday of Edgar Lederer. PMID- 3042796 TI - High-performance liquid chromatography in protein sequence determinations. AB - The use of reversed-phase high-performance liquid chromatography (RP-HPLC) for the determination of protein sequences is reported. Topics considered include the peptide separation of endoprotease digestion mixtures, the application of HPLC peptide mapping as an efficient system to check the accuracy of an assumed protein sequence, obtained indirectly by DNA sequencing and the use of HPLC for amino acid analysis in the Edman sequence strategy. The use of RP-HPLC for an unconventional sequence strategy is demonstrated; HPLC exopeptidase mapping appears to be particularly useful as a future technique for small terminal sequence analysis. Finally, the coupling of HPLC with fast atom bombardment mass spectrometry is discussed. PMID- 3042797 TI - Synthetic peptide substrates as models to study a pro-ocytocin/neurophysin converting enzyme. AB - The selectivity and mechanism of processing at paired basic amino acids in hormone precursors was studied on several analogues of the (1-20)-aminoterminal domain of the ocytocin/neurophysin precursor in a cleavage assay by an endoprotease partially purified from bovine pituitary secretory granules. Peptide analogues with amino acid substitutions in, and around, the basic doublet were synthesized and used as substrates. The data obtained demonstrate the strict requirement of the processing enzyme for basic amino acids in tandem within a possibly preferred conformation which may be highly conserved in the aminoterminal domain of this hormone precursor. PMID- 3042798 TI - Biomedical applications of chromatographic fraction containing trehalose dimycolate in squalane emulsion. AB - Trehalose dimycolate extracted from mycobacteria is a potent immunomodulator. Incorporation of trehalose dimycolate in a squalane-in-water emulsion leads predominantly to the formation of vesicular structures, which are observable by electron microscopy. The interaction between vesicles of trehalose dimycolate and the immunocompetent cells results in an enhancement of the host defence mechanisms and induction of non-specific resistance against viral, parasitic, and bacterial pathogens and certain tumors. A brief review of the pertinent observations is presented. PMID- 3042799 TI - Investigation of the lipids of saprophytic mycobacteria in the U.S.S.R. AB - Three trends in the investigations of the specific lipids of rhodococci and related microorganisms are reflected: isolation and structural determination of new complex lipids, elucidation of the role of specific lipids in the cells and application of lipid composition for diagnostic studies of rhodococci and related organisms. Two groups of peptidolipids, differing in chromatographic mobility and peptide chain structure, have been found in the cells of Rhodococcus erythropolis. The compounds within each group differ in acyl moieties. Three peptidolipids of the polar group include glucose, so they are peptidoglycolipids. Glycolipids are represented by trehalose derivates. Trehalose dimycolate (cord factor) is dominant. Data concerning the role of lipids in typical features of rhodococci as the lipophilic cell wall, the ability to up-take a hydrophobic substrate and the resistance to influences from outside are given. Examples of application of mycolic acid composition in taxonomy of rhodococci and morphologically similar bacteria are given. PMID- 3042800 TI - Concerning hydrolysis of mycolate esters, of phthiocerol dimycocerosates and of related mycobacterial lipids: an anecdotal account. AB - We review the experimental difficulties that have been encountered in hydrolyzing mycolic acid esters, their beta-O-substituted analogues, permethylated cord factor, phthiocerol dimycocerosates and similar mycobacterial lipids. Hydrolysis of the beta-O-substituted methyl mycolates is invariably sluggish and is accompanied by considerable beta-elimination to generate mycolenoic acids. Historical evidence for this often undesirable side reaction is presented. Improvements in methodology are described in which hydrolysis is promoted and beta-elimination is minimized. The reaction systems developed were found applicable to hydrolysis of the quite inert phthiocerol dimycocerosates. Permethylation of the recovered phthiocerol provides an excellent derivative for mass spectrometric analysis to define its complete structure. PMID- 3042801 TI - The vitamin K dependent reaction. AB - The vitamin K's are 2-methyl-1,4-naphthoquinones. The vitamin is required for the post-translational gamma-carboxylation of glutamyl residues in precursor polypeptides. The vitamin K step in this carboxylation, however, requires not the quinone but the hydroquinone plus oxygen. Thus, the vitamin K-dependent step is a "mixed function" oxidation requiring a reducing compound plus molecular oxygen to provide a form of oxidant (e.g., a free radical, a hydroperoxide) capable of abstracting a particular, slightly labile hydrogen from a glutamyl residue, leaving this position free to accept a carbon dioxide molecule. This oxidation appears similar to that of other mixed function oxidants such as cytochrome P450 plus oxygen, ascorbic acid (with traces of ferrous iron) plus oxygen, ferrous iron plus oxygen, and a number of other systems which function in a wide variety of oxidation. Inhibition by spin-trapping agents suggests a free radical step in the vitamin K hydroquinone-dependent reaction, similar to other mixed function oxidations. PMID- 3042802 TI - [In honor of Prof. Edgar Lederer: chromatography memoirs 1950-1960]. PMID- 3042803 TI - [Relationships between physics and medicine]. PMID- 3042804 TI - [Clinical aspect of atrial fibrillation]. PMID- 3042805 TI - Neurobehavioral effects of central nervous system prophylactic treatment of cancer in children. AB - This article reviews 41 studies of the effects of prophylactic CNS treatment on the neurobehavioral development of children with cancer. This research is classified according to studies of (a) children in treatment; (b) long-term survivors; and (c) longitudinal follow-ups of children from the time of diagnosis. Studies vary considerably in design, sample, and outcome variables, so firm conclusions regarding the morbidity of CNS prophylaxis are not currently possible. However, the studies do suggest that CNS prophylaxis does impair cognitive development, particularly when cranial radiation therapy is part of the treatment. There is also evidence of greater impairment in younger children and some suggestion of more frequent impairment of non-language skills relative to language skills. The possible relationships among age, radiation, and non language cognitive skills may be linked to disruption of white matter CNS structures apparent on autopsy and cerebral tomography following treatment. PMID- 3042806 TI - Collagen measured in primary cultures of normal rat hepatocytes derives from lipocytes within the monolayer. AB - The cellular origin of hepatic collagen is under active investigation. Several recent studies using cells in primary culture suggest that hepatocytes are the source of much of the collagen in normal rat liver. In view of other data indicating that lipocytes produce substantial amounts of this protein, we have reexamined collagen biosynthesis in hepatocyte cultures that have been carefully characterized with respect to the presence of lipocytes. We find that routinely prepared hepatocyte isolates contain, by number, approximately 10% lipocytes. Lipocytes in early culture are difficult to visualize by phase-contrast microscopy but after 4 d proliferate and eventually replace the parenchymal cells. The size of the lipocyte subpopulation in these cultures correlates positively with collagen production. Similarly, removal of lipocytes by further processing of the initial hepatocyte isolate significantly reduces collagen production. Moreover, the only cells within hepatocyte cultures that display type I collagen by immunohistochemistry are lipocytes. We conclude that lipocytes are the principal source of collagen in primary hepatocyte cultures. The findings indicate also that these cells are the previously described "fibroblast" that appear in relatively long-term hepatocyte cultures. PMID- 3042807 TI - Release of carcinoembryonic antigen from human colon cancer cells by phosphatidylinositol-specific phospholipase C. AB - Carcinoembryonic antigen (CEA) is released from colon cancer cells into the circulation where it is monitored clinically as an indicator of the recurrence or progression of cancer. We have studied the mechanism of CEA membrane attachment and release using the human colonic adenocarcinoma cell line LS-174T, specimens of human colon cancers, and serum from colon cancer patients. CEA release by cells in vitro and in vivo is associated with the conversion of CEA from a membrane-bound, hydrophobic molecule to a soluble, hydrophilic form with no apparent decrease in molecular mass. When LS-174T cell membranes were incubated with various buffers, proteases, and phospholipases, the only agents that released CEA and converted it to the hydrophilic form were preparations of phosphatidylinositol-specific phospholipase C (PI-PLC). Both [3H]ethanolamine and [3H]palmitate could be incorporated metabolically into CEA but only palmitate was released by treatment with PI-PLC, consistent with the presence of a glycosyl phosphatidylinositol linkage. PI-PLC treatment also release significant quantities of CEA from living monolayers and from seven human colon cancer specimens. These experiments suggest that cellular CEA is anchored to membranes by a covalent linkage to a membrane phosphatidylinositol molecule, and that an endogenous phospholipase may be important for releasing CEA in vitro and in vivo. PMID- 3042808 TI - Demonstration by in situ hybridization of the zonal modulation of rat liver cytochrome P-450b and P-450e gene expression after phenobarbital. AB - The various physiological processes that constitute liver function are compartmentalized within the hepatic acinus. The molecular mechanisms modulating the development and maintenance of this hepatocyte heterogeneity have not been defined. The objective of this study was to determine whether transcriptional or posttranscriptional zonal modulation of cytochromes P-450b,e gene expression was responsible for the heterogeneous induction of the P-450 proteins, which is observed after phenobarbital (PB) administration. The exact localization in liver tissue of hepatocytes responding to PB with induction of either P-450b,e mRNA or proteins was established by in situ hybridization and by immunofluorescence, respectively. As demonstrated by quantitative assessment of autoradiographs of approximately 20 hepatocytes located between a terminal portal venule and a hepatic venule, PB induced the P-450b,e mRNA up to sixfold in the 12-15 hepatocytes located closer to the hepatic venules (zones 2 and 3). In contrast, there was only a twofold induction in the 4-6 hepatocytes surrounding the terminal portal venules (zone 1). Quantitative immunofluorescence using an MAb showed that the acinar distribution of PB-induced P-450b,e proteins was similar to that of the mRNA. This combined approach indicated that, most likely, an increased rate of transcription of cytochromes P-450b,e genes in hepatocytes of zones 2 and 3 concomitantly, with a relative lack of activation, or repression, of these genes in hepatocytes of zone 1, were responsible for the heterogeneous phenotype observed after PB administration. Therefore, modulation of gene expression among hepatocytes of the liver acinus is one mechanism by which the functional heterogeneity of hepatocytes is attained. Experiments in which the induction of cytochromes P-450b,e genes was studied after administration of either PB or para-hydroxyphenobarbital, a main hepatic metabolite of PB, suggested that the species involved in the inductive process is the parent PB molecule rather than para-hydroxyphenobarbital. PMID- 3042809 TI - Synovial osteochondromatosis of the temporomandibular joint. An historical review with presentation of 3 cases. AB - Loose bodies in joints have long held the fascination of surgeons and their recognition clearly enjoys a most distinguished antiquity (Pare, 1558; Haller, 1764; Barwell, 1876; Halstead, 1895) despite the rarity of their occurrence. This paper presents an historical review of the subject in conjunction with a report of three cases operated upon by one of the authors (JdeBN). The results have been critically reviewed by pathologists with a catholicity of experience in the field. The condition is singularly uncommon in the jaw joint, and must rank with synovial cyst and para-articular chondroma as an unusual cause of a swelling of firstly the temporomandibular joint and secondly the parotid gland. Earlier workers recognized the value of comparative pathology to illustrate the nature of a genus of tumours and it is educative to read the following: "A good physiological type for the loose cartilaginous bodies which infest joints is furnished by the temporomandibular joint of the skate. A recess communicating with this articular cavity usually contains a collection of smooth cartilaginous bodies, in contour and size like melon seeds." Bland-Sutton (1907). PMID- 3042810 TI - Resiliency: research and practical implications for pediatricians. PMID- 3042811 TI - Detection of high-risk groups and individuals for periodontal diseases. Systemic predisposition and markers of general health. AB - The evidence for systemic predisposition to periodontal diseases is reviewed in relation to cellular and humoral immunity, drug therapy, diet and nutrition and stress. It is concluded that, apart from defects of polymorphonuclear leukocytes (PMN) and Ehlers-Danlos Syndrome, little firm evidence exists for other diseases, though insulin-dependent diabetes and acquired immune deficiency syndrome (AIDS) may accelerate and/or potentiate the damage of existing disease. The precise role of drugs, diet and nutrition and stress remain to be elucidated, but recent advances in these areas offer the prospect of assessing risk using carefully controlled studies. PMID- 3042812 TI - A comparison of 4 methods of data presentation for lysosomal enzyme activity in gingival crevicular fluid. AB - In previous studies, we have emphasized the importance of considering the methods used for analysis of gingival crevicular fluid (GCF). This study evaluated 4 different approaches for data presentation of lysosomal enzyme activity in GCF. GCF was collected from patients displaying at least 2 mm of clinical attachment loss at a minimum of 3 sites in the mouth (DA), and patients who did not display clinical attachment loss of 2 mm or more at any site in the mouth (DI), during a 3-month interval following entry into a longitudinal trial. GCF was collected by the timed intrasulcular placement of precut filter paper strips. 16 to 28 individual GCF samples were collected from each patient. The lysosomal enzymes studied were B-glucuronidase (BG) and arylsulfatase. The mean values for the DA and DI groups at baseline and 3 months are reported. The results indicate that when the data is expressed as total enzyme activity (unit activity) per 30-s collection (UA) or UA x GCF volume (microliter) per mm of probing depth, the DA group demonstrated significantly greater mean values than the DI group at baseline and 3 months. In contrast, when the data was expressed as concentration (UA/microliter), or UA per mm of probing depth, differences between the DA and DI groups were observed only at the 3-month evaluation. The difficulty in using concentration when reporting GCF lysosomal enzyme activity is emphsized by comparison of the data from the DA group and the high and low enzyme activity subsets of the DI group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3042813 TI - Combined antibiotic (metronidazole) and mechanical treatment effects on the subgingival bacterial flora of sites with recurrent periodontal disease. AB - 5 patients in maintenance, 1-3 years after periodontal therapy who showed sites with reinfected pockets and bleeding despite regular recall visits were selected. Darkfield microscopy from 3 sites in each patient showed an average of 41% spirochetes and 21% motile rods. Probing depths ranged from 7 to 9 mm and loss of clinical attachment from 6 to 13 mm in these sites. The patients were given 3 times 250 mg/day of metronidazole (Flagyl) for 10 days. Darkfield microscopy and microbiological cultures of the subgingival plaque were performed twice prior to the study, at the end of the medication and after 3 weeks, 3 and 6 months. The plaque and gingival indices, probing depth and loss of clinical attachment were recorded. During the medication and at 3 and 6 months, the teeth were scaled and root planed. The samples were obtained with 3 paper points and cultured anaerobically in the glove box on non-selective and selective media and representative bacterial colonies identified by aerobic growth, gram stain and rapid biochemical tests. Presumptive pathogenic micro-organisms including Bacteroides were identified. The % of spirochetes, motile rods and non-motile bacteria were enumerated by darkfield microscopy. The clinical results show that administration of metronidazole and repeated root planing significantly decreased gingival inflammation, probing depth and loss of clinical attachment in reinfected sites. After treatment, these sites harbored significantly less spirochetes and more non-motile bacteria, while motile rods tended to return to baseline levels with time. The combined antibiotic and mechanical therapy resulted in a statistically significant decrease of gram-negative rods, Fusobacteria and Bacteroides gingivalis over 6 months.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3042814 TI - Treatment of intraosseous periodontal defects with a combined adjunctive therapy of citric acid conditioning, bone grafting, and placement of collagenous membranes. AB - A total of 25 proximal, intraosseous periodontal defects were treated in 21 adult patients. A 3-prong adjunctive, regenerative treatment approach was used. The treatment included (1) citric acid conditioning of the root surfaces, (2) grafting of particles of decalcified, freeze-dried homologous bone, and (3) placement of freeze-dried, homologous dura mater sheets between the replaced surgical flaps and the tooth surfaces. The results, as evaluated by probing attachment and probing bone level measurements, during 1 year of observation, demonstrated limited improvements of the treated defects. The limited results were similar to previous observations in our clinics following treatment of intraosseous defects using different treatment modalities. It appears that new treatment approaches need to be sought to accomplish clinically significant and predictable regeneration in proximal, intraosseous periodontal defects. PMID- 3042815 TI - Methotrexate in rheumatoid arthritis. PMID- 3042816 TI - Methotrexate in psoriasis: revised guidelines. PMID- 3042817 TI - Lipomembranous changes in chronic panniculitis. AB - The clinical and histopathologic findings in 13 patients with lipomembranous changes in the subcutaneous adipose tissue as part of the inflammatory reaction are presented. Nine patients had clinical evidence of vascular disease and four had clinical evidence of connective tissue disease. Histopathologic evidence of endarteritis obliterans, venous stasis, and hemorrhage was present in more than half the patients, and the clinical lesion of liposclerosis was frequently present. These findings suggest that the histologic changes of lipomembranous panniculitis may be the result of an inflammatory reaction in patients who have the liposclerosis of venous insufficiency with connective tissue disease or previous leg ischemia or both. PMID- 3042818 TI - Dermatologic therapy: December 1986 to December 1987. AB - In this article I review significant therapeutic advances reported in the English literature from December 1986 to December 1987. Readers should review the original articles in toto before attempting any new experimental or controversial therapy summarized. PMID- 3042819 TI - Pemphigoid-like bullous eruption related to ibuprofen. AB - We report two patients who had a bullous eruption limited to the lower extremities when first seen by us. Clinically the eruption resembled limited bullous pemphigoid. In both patients the onset of the eruption was associated with ibuprofen therapy, and lesions promptly resolved after discontinuing the ibuprofen. This type of presentation has not been reported previously as an adverse effect of ibuprofen. Bullous reactions to nonsteroidal anti-inflammatory agents are briefly reviewed. PMID- 3042820 TI - Dermal, subcutaneous, and tendon xanthomas: diagnostic markers for specific lipoprotein disorders. AB - Many patients with lipoprotein disorders are at increased risk for the development of premature atherosclerosis and, less commonly, other disorders that cause systemic morbidity. In some of these patients, xanthomas also develop and provide cutaneous markers for the lipoprotein disorder. As advances in molecular biology refine our understanding of lipoprotein metabolism, it has become increasingly clear that several types of xanthomas are associated with specific disease states. This article presents a differential diagnosis of xanthomas that incorporates contemporary thinking about lipoprotein disorders and focuses on the relationship between abnormalities in lipoprotein metabolism, content, or structure and the development of specific xanthomas. PMID- 3042821 TI - An analysis of mental health research with American Indian youth. AB - A comprehensive search and review of literature, documents, publications and other material written since 1970, relevant to American Indian mental health research, was completed in order to generate a list of research and of training needs. A content analysis of this literature enabled a synthesis which describes important biopsychosocial issues faced by American Indian communities, gaps in past and current research efforts, specific problems in past research, and recommendations in each of these areas with regards to future research possibilities and needs. The portion of the analysis presented in this paper deals with American Indian infants, preschoolers, children and adolescents. In order of presentation, the specific issue domains dealt with include: otitis media, fetal alcohol syndrome, abuse and neglect, failure-to thrive/autism/enuresis, which are examined together (early development) in terms of research gaps; and neurosensory disorders/developmental disabilities/handicapping conditions/school-related problems, foster care and adoption, self-concept/identity, conduct disorders/delinquency, drug and alcohol use, and suicide and depression, which are examined (school-age children and adolescents) in relation to research gaps and needs. PMID- 3042822 TI - Cowside antibiotic residue testing. AB - Detectable concentrations of antibiotic residues in milk supplies are illegal. They interfere with manufacturing of some dairy products, may cause hypersensitivity or resistance to drug therapy in humans, and are perceived by consumers as undesirable. Antibiotics are used for intramammary, intramuscular, oral, or reproductive therapy to counter acute or subacute diseases. Residues can result for many reasons, including poor records of treatment, failure to observe recommended label withdrawal time, prolonged drug clearance, treated animal identification problems, products not used according to label directions, lack of advice on withdrawal period, and others. Several effective and useful on-farm residue screening tests are available and should be used to monitor cows treated with an antibiotic for any reason. Herds that use drugs without label directions or that do not follow label directions should find these tests valuable. Producers must understand how to interpret the results of these tests. Also, dairy farmers must recognize if the selected test method is capable of detecting those antibiotics in use and what are to be monitored. PMID- 3042825 TI - Dental school adaptation for a paraplegic dental student. PMID- 3042824 TI - Attachment of Bacteroides gingivalis to collagenous substrata. AB - The ability of Bacteroides gingivalis 381 to attach to hydroxyapatite (HA) beads, treated with either human type I or type IV collagen, or to particles of bovine bone collagen was studied. All preparations were blocked with human albumin prior to being incubated with 3H-thymidine-labeled B. gingivalis 381 cells. The presence of collagen on HA surfaces (C-HA) significantly promoted attachment of the organism. HA treated with Type IV collagen bound B. gingivalis cells more effectively than did HA treated with type I collagen. Attachment of two additional strains of B. gingivalis to HA was also promoted by collagen. Binding to type I or type IV C-HA occurred rapidly, and equilibrium was attained within 45 min. B. gingivalis 381 cells also bound to particles of bovine bone collagen, and this appeared to be biphasic. Heating the bacteria abolished their ability to bind to C-HA. Attachment of B. gingivalis 381 cells to HA treated with type I collagen was strongly inhibited by the presence of soluble type I or type IV collagen, or gelatin, but not by the presence of human albumin, salivary proline rich protein 1, or saliva. Human serum, fibronectin, fibrinogen, certain protease inhibitors, and some peptides were also inhibitory. 3H-fibronectin bound to bovine bone collagen particles and blocked the attachment of 14C-B. gingivalis cells. Mild trypsin treatment of the fibronectin-collagen complex restored its ability to promote 14C-B. gingivalis attachment concomitant with the loss of 3H fibronectin. We suggest that elevated levels of proteases in the gingival sulcus, such as are associated with poor oral hygiene and gingivitis, might remove fibronectin and expose collagen molecules in the basement membrane, thereby promoting the attachment of B. gingivalis cells and facilitating their invasion into gingival tissues. PMID- 3042823 TI - Correlations between gingival crevicular fluid enzymes and the subgingival microflora. AB - Bacteroides gingivalis is a Gram-negative micro-organism implicated in the pathogenesis of adult periodontitis and producing relatively large amounts of specific enzymes. In the present study, subgingival samples taken from adults with moderate periodontitis were examined for the presence and relative amounts of enzymatic activity toward certain substrates. Enzyme levels were then correlated with clinical periodontal indices and microbiological analysis of subgingival plaque, including darkfield microscopy for bacterial morphotypes and immunofluorescence microscopy for B. gingivalis and Bacteroides intermedius. The results of this study indicate a significant positive correlation between levels of enzyme capable of degrading N-benzoyl-D,L-arginine-beta-naphthylamide hydrochloride, and subgingival B. gingivalis (r = 0.55). There was a much lower correlation coefficient between this enzyme activity and subgingival B. intermedius (r = 0.26). Statistically significant (p less than 0.01) positive correlations were also demonstrated between total bacterial cell counts and levels of enzymatic activity against N-benzoyl-D,L-arginine-beta-naphthylamide hydrochloride (r = 0.76), N-carbobenzoxy-glycyl-glycyl-L-arginine-beta naphthylamide hydrochloride (r = 0.72), and glycyl-L-proline-4-methoxy-beta naphthylamide hydrochloride (r = 0.72), and glycyl-L-proline-4-methoxy-beta naphthylamide hydrochloride (r = 0.69). There were significant differences in the levels of these three enzymatic activities between sites exhibiting various degrees of clinical severity of gingival inflammation and harboring various proportions of B. gingivalis. The data from this study indicate that measurement of specific enzymatic activities in subgingival samples can be useful in the diagnosis of B. gingivalis-associated periodontitis. PMID- 3042827 TI - Prognostic assessment of the acute complications of bone marrow transplantation requiring intensive therapy. AB - Patients with bone marrow transplant may present with acute, life-threatening complications which frequently (40% of our cases) require intensive care unit treatment and result in an increased mortality (76% in this series). In an attempt to reach a more objective prognostic assessment, we have analyzed those factors related to the worst outcome in the 25 patients with bone marrow transplant admitted into our intensive care unit. Respiratory failure was the most frequent complication (72%), with an 83% mortality. Graft-versus-host disease and neutropenia led to a greater number of infectious complications with a poor outcome. Failure of more than three organ systems, septic shock and mechanical ventilation were statistically associated with mortality (p less than 0.05), and all patients who required mechanical ventilation for more than seven days or needed intensive therapy for more than 10 days died. The presence of septic shock, multisystem failure and severe neutropenia on admission should be considered as initial indicators of a poor prognosis. More than 7 days of mechanical ventilation and an intensive care unit stay of more than 10 days could be critical points in the reassessment of the intensity and prolongation of treatment. PMID- 3042826 TI - Pressure-volume curves, static compliances and gas exchange in hyaline membrane disease during conventional mechanical and high-frequency ventilation. AB - Eight premature infants with hyaline membrane disease needing artificial ventilation were studied at a mean age of 26.5 h. After a preparative phase they were randomly assigned either first to conventional mechanical ventilation (CMV; delivered by a Siemens Servo 900 C), followed by high-frequency ventilation (HFV; delivered by Percussionaire VDR 1 at 10 Hz) or vice versa, each period lasting 4 h. At the end of each period, arterial blood gases, lung volumes and alveolar pressures (Palv) during CMV or HFV and pressure-volume (P-V) curves of the total respiratory system were determined. Expiratory volumes were measured spirometrically, Palv by the clamping method, and the P-V curve was constructed by the syringe method. Single point static compliance at end-inspiration was higher during HFV (0.40 +/- 0.10 vs. 0.32 +/- 0.08 ml/cmH2O.kg-1; p = 0.02), whereas at end-expiration no difference was noted. Two points static compliances were also better during HFV than during CMV (0.32 +/- 0.08 vs. 0.24 +/- 0.06 ml/cmH2O.kg-1; p = 0.01). Static compliances derived from the steepest part of the inflation limb of the P-V curve were 0.55 +/- 0.12 after CMV and 0.50 +/- 0.12 ml/cmH2O.kg-1 after HFV (n.s.). Compared to CMV, HFV resulted in similar oxygenation and CO2-elimination at equal mean lung volumes, but at significantly lower mean Palv. It is concluded that recruitment of lung volume is achieved with less static recoil pressure by HFV. These findings are explained by differences in inspiration allowing more time for volume recruitment during HFV. PMID- 3042828 TI - Ventilatory stimulus suppression in intrinsic PEEP patients. PMID- 3042830 TI - Changes of right ventricular function with positive end-expiratory pressure (PEEP) in man. AB - The side effects of positive pressure ventilation on cardiovascular function are well known. However, in most clinical studies its influence on left ventricular function was examined. In the present study right ventricular (RV) performance was studied in 13 patients undergoing coronary artery bypass grafting during mechanical ventilation with and without positive end-expiratory pressure (PEEP). In the majority of patients (10/13), PEEP caused a decrease in RV end-diastolic volume (by 18%) whereas RV ejection fraction did not change. In the remaining 3 patients, end-diastolic and end-systolic RV volumes increased by 25% and 50% respectively and ejection fraction decreased by 29%. These results suggest that PEEP can affect RV function in two different ways: first, in the majority of patients studied, PEEP exerted an unloading effect on the RV whereas second, in 3 patients RV dilatation and a decrease in ejection fraction was observed. PMID- 3042831 TI - Right ventricular ejection fraction measurement in moderate acute respiratory failure (ARF). Effects of PEEP. AB - Eight patients mechanically ventilated for acute respiratory failure were submitted to increasing levels of PEEP, from 0 to 15 cm H2O. Right ventricular ejection fraction (RVEF) and end-diastolic volume index (RVEDVI) were measured using the fast response thermistor Swan-Ganz catheter. PEEP induced a linear decrease of cardiac index, while the pulmonary artery pressure increased. In three patients (group A) with a RVEDVI larger than 120 ml at ZEEP, RVEF decreased and RVEDVI increased with PEEP. In the other five patients (RVEDVI less than 120 ml, group B), RVEF was unchanged and RVEDVI decreased at PEEP 15 cm H2O. This study suggest that RV changes induced by PEEP are probably a function of the initial RVEF and RVEDVI. PMID- 3042829 TI - The right ventricle and critical illness: a review of anatomy, physiology, and clinical evaluation of its function. AB - This paper reviews right ventricular anatomy and physiology in the critically ill patient. The role of right ventricular function during acute pulmonary artery hypertension and the effect of acute myocardial injury upon right ventricular performance are examined. Clinical methods of assessing right ventricular function at the bedside in acutely ill patients are critically reviewed. PMID- 3042832 TI - The influence of catecholamines on right ventricular function in septic shock. AB - Catecholamines play an important role in the treatment of septic shock. Not much is known about their effects on right ventricular function. In this paper the available data on the effects of different catecholamines on right ventricular dysfunction complicating septic shock are reviewed. PMID- 3042833 TI - Right ventricular function and cardiac surgery. PMID- 3042834 TI - Methodological considerations in the evaluation of the convergence of psychiatric diagnoses and parent-informant checklists. AB - Psychiatric diagnoses and objective parent checklists are alternative ways to describe child adjustment problems. There has recently been interest in evaluating the degree of agreement or convergence between these sources of information. This paper addresses three issues neglected by researchers in this area. The appropriateness of the use of indices of sensitivity and specificity to describe the correspondence of diagnoses and checklist scores is questioned. Implications of failure to consider the reliability of diagnoses in interpreting diagnosis-checklist agreement are discussed. Also, possible parameters of diagnosis-checklist agreement that should be identified by researchers are reviewed. Suggestions for improving research in this area are offered. PMID- 3042836 TI - Endothelium-derived relaxing factor. AB - This article reviews what is known of endothelium-derived relaxing factor and its possible physiologic and pathophysiologic roles. This relaxing factor is now thought to be nitric oxide or a ready source of it. It acts as an endogenous nitrovasodilator, stimulating soluble guanylate cyclase to increase cyclic guanosine monophosphate (GMP) levels in vascular smooth muscle and platelets, with consequent relaxant and anti-aggregatory effects (predominantly when stimulated through receptor-operated channels). Its actions are thus synergistic with those of cyclic adenosine monophosphate (AMP)-mediated stimulation (for example, adenosine, prostacyclin). Endothelium-derived relaxing factor is unstable and is thought to act only very locally in vivo. Its release is continuous in the basal state and is stimulated by a number of neuropeptides and by agents released during platelet activation and thrombosis--with large differences in activity among different vessels. Endothelium-derived relaxing factor activity is also flow related, thereby coordinating vasomotor behavior in an intact vascular tree in response to changes in flow. Endothelium-derived relaxing factor activity is reduced in several pathologic states, including atherosclerosis. PMID- 3042835 TI - Intravenous recombinant tissue plasminogen activator (rt-PA) and urokinase in acute myocardial infarction: results of the German Activator Urokinase Study (GAUS). AB - The effects of recombinant tissue plasminogen activator (rt-PA) and urokinase on patency and early reocclusion of infarct-related coronary arteries were investigated in a single blind, randomized multicenter trial in 246 patients with acute myocardial infarction of less than 6 h duration. Both 70 mg of single chain rt-PA with an initial bolus of 10 mg and 3 million units of urokinase with an initial bolus of 1.5 million units were given intravenously over 90 min. The first angiographic study at the end of the infusion revealed a patent infarct related artery (Thrombolysis in Myocardial Infarction trial [TIMI] grade 2 or 3) in 69.4% of 121 patients given rt-PA versus 65.8% of 117 patients given urokinase (p = NS). Among patients treated within 3 h from symptom onset a patent infarct related artery was found in 63.9% of 72 patients given rt-PA versus 70% of 70 patients given urokinase (p = NS). There were five cardiac deaths in each group and one fatal intracranial hemorrhage in the rt-PA group. The in-hospital reinfarction rate was 8.9% versus 13.2% for patients treated with rt-PA and urokinase, respectively. There was no difference in left ventricular function at baseline and follow-up catheterization studies. Both drugs were well tolerated and there was no significant difference in cardiovascular or bleeding complications between the two groups. It is concluded that rt-PA and urokinase in the dosages used provide similar efficacy and safety in the treatment of acute myocardial infarction. Reocclusion during the first 24 h may be less frequent after urokinase treatment. PMID- 3042837 TI - Analysis of bronchoalveolar lavage in allergic bronchopulmonary aspergillosis: divergent responses of antigen-specific antibodies and total IgE. AB - Bronchoalveolar lavage (BAL) was performed in eight patients with allergic bronchopulmonary aspergillosis (ABPA) at a time when chest roentgeongraphy did not reveal an infiltrate, and respiratory status was stable. BAL was tolerated well by all patients with only one patient experiencing mild wheezing. BAL fluid recovery averaged 40%, and total cells/lavage were 22.3 x 10(6) (range 3.5 to 49.5 x 10(6)). Cell viability, as determined by trypan blue exclusion, averaged 48% (range 34% to 60%). Mean values for cellular elements were macrophages, 62%; epithelial cells, 12%; lymphocytes, 16%; neutrophils (PMN), 4%; and eosinophils, 6%. Isotypic antibodies to Aspergillus fumigatus (Af) in BAL and serum were detected by an amplified indirect ELISA. Antibodies to Af in BAL expressed as optical density/albumin (milligrams per milliliter) were compared to BAL from six nonatopic patients. IgE-Af and IgA-Af in BAL were elevated in patients with ABPA compared with six nonatopic patients. The ratios of Ig-Af in BAL to peripheral blood in patients with ABPA were 48 (range 18 to 75) for IgE-Af, 96 (range 37 to 159) for IgA-Af, and 0.94 (range 0.24 to 1.40) for IgG-Af, suggesting local production of IgE-Af and IgA-Af in the bronchoalveolar compartment. Total serum IgE correlated directly with IgE-Af in BAL (rs = 0.67; p less than 0.02). However, the ratio of total BAL IgE/albumin divided by total serum IgE/albumin was 0.93 +/- 0.94, suggesting that the bronchoalveolar compartment is not the source of the significant elevations in total serum IgE in ABPA. PMID- 3042838 TI - Immunologic response to Faenia rectivirgula (Micropolyspora faeni) in a dairy farm family. AB - In the present study, cellular and humoral responses to Faenia rectivirgula antigens were evaluated in seven subjects, members of a family who lived and worked on a dairy farm. Four subjects had clinical features of hypersensitivity pneumonitis after exposure to moldy hay. The other three subjects had no clinical disease in spite of similar exposure. Although serum precipitins were found in most subjects, a biotin-avidin-linked immunosorbent assay revealed high levels of F. rectivirgula-specific antibodies only in the symptomatic subjects. In addition, numerous precipitin arcs were present in the sera of the symptomatic but not the asymptomatic subjects by antigen-antibody crossed immunoelectrophoresis. No clear distinction between symptomatic and asymptomatic subjects could be made on the basis of lymphocyte phenotype studies, and antigen induced lymphocyte transformation was not detected in any subjects. The results indicate that F. rectivirgula-specific antibody levels as detected by biotin avidin-linked immunosorbent assay and by the presence of precipitin arcs in crossed immunoelectrophoresis may differentiate symptomatic and asymptomatic farmers. PMID- 3042839 TI - Soybean flour asthma: detection of allergens by immunoblotting. AB - A 43-year-old woman developed asthma 6 years after beginning work in a food processing plant in which soybean flour was used as a protein extender. Symptoms of sneezing, coughing, and wheezing would begin within minutes of exposure to soybean flour and resolve 2 hours after exposure ceased. Skin tests were positive to a soy extract prepared from the flour. Airway hyperreactivity was confirmed by a positive bronchial challenge to methacholine. Bronchial challenge with soybean flour produced an immediate increase in specific airway resistance from 5.0 to 22.7 L. cm of H2O/L/sec. There was no response to challenge with lactose. The patient's allergic response to soy-flour extract was further characterized by several immunologic methods. IgE binding to soy-flour protein by direct RAST was 5.98 times that of a normal control serum. The soy-flour extract was separated by dodecyl sulfate-polyacrylamide gel electrophoresis. Twenty-four protein bands were detected in the crude soy-flour extract. After immunoblotting and subsequent autoradiography, nine proteins with molecular weights ranging from 54,500 to 14,875 were found. Cross-reactivity studies with other legumes demonstrated apparent immunologic identity between a component in green pea extract and a soybean protein with a molecular weight of 17,000. The clinical significance of this cross-reactivity is not known. We conclude that in this case of occupational asthma to soybean flour, multiple allergens were involved. Immunoblotting may be useful in identifying the allergens involved in occupational asthma. PMID- 3042840 TI - Inhaled corticosteroids--their present and future role in the management of asthma. PMID- 3042841 TI - High-dose naloxone in older normal subjects: implications for Alzheimer's disease. AB - The behavioral and cognitive effects of naloxone HCl, in doses of 5 micrograms/kg, 0.1 mg/kg, and 2.0 mg/kg administered as an IV bolus, were assessed in a double-blind, placebo-controlled, randomized study of eight normal subjects ranging in age from 44 to 74 years (mean 63). Naloxone produced mild behavioral effects with slight cognitive impairment after the 2.0 mg/kg dose only. The threshold, dose dependency, characteristics, and magnitude of these behavioral effects were similar to what has previously been reported in young normal subjects, but markedly different from those observed in patients with dementia of the Alzheimer type (DAT) matched in age to the current study sample. These data suggest that the metabolic fate of naloxone is not substantially affected by age within the range studied. The findings of this study provide further support for a role for endogenous opiate systems in the modulation of behavior and cognition, and suggest that the unusual behavioral sensitivity of patients with DAT to naloxone cannot be accounted for by the effect of age. PMID- 3042842 TI - Self-rated depression scales and screening for major depression in the older hospitalized patient with medical illness. AB - Until now, no self-rated depression scale had been validated as a screening measure for major depression in the older patient hospitalized with medical illness. The present report establishes the validity of two brief, easily administered depression screening tests, the Geriatric Depression Scale (GDS) and the Brief Carroll Depression Rating Scale (BCDRS), in this population. Structured psychiatric interviews were performed and self-rated depression measures administered to 128 men, aged 70 and over, consecutively admitted to medical and neurological services of a VA hospital. The GDS and BCDRS were both shown to have high sensitivity and specificity for detecting major depression in this setting. Optimal cut-off scores determined by the receiver operating curve characteristics of these tests were 11 for the GDS and 6 for the BCDRS. At a cutoff score of 11, the GDS had a sensitivity of 92%, a specificity of 89%, and a negative predictive value of 99%; lowering the break point to 8 did not increase sensitivity. At a cutoff score of 6, the BCDRS achieved a 100% sensitivity, 93% specificity, and 100% negative predictive value. Whether clinicians decide to implement either of these depression screens in their practice will depend to a large degree on the importance ascribed to the detection of these disorders and on attitudes toward the benefits of treatment. PMID- 3042843 TI - Clinical correlates of bacteremia in a Veterans Administration extended care facility. AB - Little is known about bacteremia in long-term care facilities. We have conducted a retrospective study during a 12-month period analyzing the clinical correlates of bacteremia in 533 chronically institutionalized, predominantly male patients, with an average age of 69 years. Thirty-four men had forty-two bacteremic illnesses during this period. The incidence rate was 0.30 episodes per 1000 patient care days, and the mortality rate was 21%. The urinary tract was the most frequently identified tissue source (56%), followed by respiratory tract (7%) and skin (7%). Providencia stuartii was the most common gram-negative organism, while Staphylococcus aureus, Streptococcus pneumoniae and enterococcus were the frequent gram-positive microbes. Gram-negative bacteremia accounted for 63% of the episodes (15% mortality rate), and gram-positive bacteremia accounted for 27% (18% mortality rate); 10% of the bacteremias were polymicrobial (25% mortality rate). Most of the isolated organisms were sensitive to available antimicrobial agents. The leading risk factor for bacteremia was an indwelling urinary catheter (odds ratio 39, 95% confidence limits 16 to 97). Patients with urinary catheters at the beginning of the study constituted only 5% of the population, but accounted for 40% of the gram-negative bacteremias during the year of observation. PMID- 3042844 TI - An evaluation of increased mortality rates in Wisconsin nursing homes. AB - We reviewed mortality data from 80 nonprofit and government-owned skilled nursing facilities (SNFs) to evaluate previously reported increases in deaths occurring in Wisconsin nursing homes since 1983. Comparing nursing home mortality data for 1982 and 1985, we found a 16.6% increase in overall nursing home mortality rates. The increased mortality rates occurred in the sample SNFs regardless of ownership, Medicare certification, bed size, metropolitan area and hospital affiliation. There were two explanations for the increased mortality rates. First, the number of residents dying within 30 days after nursing home admission increased 59%. The majority of these short-lived residents had been discharged from hospitals indicating a transfer of terminally ill patients into nursing homes just prior to death. Second, there was a 27% increase in the mortality rate of residents living in the nursing home for 1 to 5 years suggesting that the population had become sicker between 1982 and 1985. These data reflect both the impact of Medicares Prospective Payment System (PPS) on the study nursing homes and an increase in the severity of illness of Wisconsin's nursing home population between 1982 and 1985. The findings document an increased role for nursing homes in caring for more acutely ill patients since the passage of the PPS, and have implications for nursing home reimbursement policies and quality of care. PMID- 3042845 TI - Acute care in chronic care settings. PMID- 3042846 TI - Elder abuse: ten years later. PMID- 3042847 TI - New instrumentation in optometric practice. PMID- 3042848 TI - Cavernous hemangioma of the retina: a neuro-oculocutaneous phakomatosis. AB - A case of retinal cavernous hemangioma is presented. It is now recognized as a phakomatosis with neuro-oculocutaneous manifestations. PMID- 3042850 TI - Comments on correspondence between Henry Cohen and Frank R. Hartman. PMID- 3042849 TI - The consummate teacher of optometry: Professor E. R. Tennant. PMID- 3042852 TI - Foreign body reactions. PMID- 3042853 TI - Diabetes and healing: a review of the literature. AB - Blood sugar control is an important factor for healing in the diabetic surgical patient. Careful assessment preoperatively of all patients for glucose is essential. A surgical zone of serum glucose should be established for diabetics, and this zone should be maintained throughout the procedure and postoperative period. Blood sugar control should be maintained by the patient throughout the healing phase, as poor control may interfere with the healing process. PMID- 3042851 TI - Interpretation of microbial bioassays using kinetic parameters. AB - A method analogous to that used for enzyme kinetics was applied to estimate maximum growth rates and oxygen depletion rates as measures of microbial toxicity. Selected toxicants including phenol, formalin and CuSO4 were tested against an axenic culture of E. coli and a mixed culture derived from soil to determine reproducibility of the test procedures. Optical density was used as a measure of growth over time. Oxygen uptake over a short term (less than 20 min.) and over extended time (greater than 4 hrs.) was also monitored. Microbial growth rate constants (g) and oxygen depletion rate constants (k) were calculated and the EC50 values for toxicity were taken as one half the maximum growth rate or oxygen depletion rates, respectively. The method proved a reasonable and reliable measure of toxicity. PMID- 3042854 TI - Susceptibility testing today: myth, reality, and new direction. PMID- 3042855 TI - Flash sterilization. PMID- 3042856 TI - Introduction of a plasmid encoding the OHIO-1 beta-lactamase to an intermediate care ward by patient transfer. AB - A prospective study of 147 intermediate care ward (ICW) patients for acquisition of gentamicin-resistant Enterobacteriaceae (GRE) was carried out. Fifty (34%) were colonized or infected with one or more strains of GRE. Fifteen of these patients and one nurse were colonized with 22 strains (including ten species) of GRE bearing identical 60 kb plasmids encoding a novel beta-lactamase determinant, OHIO-1 and ANT(2"). Analysis of the time course of colonized patients on the ICW revealed one probable episode of cross-transmission. Five colonized patients had been residing in the ICW from one to four months prior to study initiation. Eight patients were admitted to the ICW from other hospital areas already colonized and one additional patient acquired colonization on the ICW from an unknown source. Thus, eight of ten patients admitted to the ICW during the prospective study were already colonized on admission to the ICW. To control this level of colonization it would therefore be necessary to direct efforts at limiting admission of colonized patients or attempting to eliminate the strain from persistently colonized patients, rather than trying to limit transmission within the ward. PMID- 3042858 TI - Sleepiness and motor vehicle accidents. PMID- 3042857 TI - Clinical pharmacology of antibiotics. Sulbactam/ampicillin. AB - Sulbactam/ampicillin appears to be effective in the treatment of gynecologic and intra-abdominal infections, and infections of skin and skin structures. However, until more data is available from well-controlled, comparative studies, it is difficult to determine the most appropriate place of sulbactam/ampicillin in the therapy of these and other infections. Sulbactam/ampicillin is similar to other agents in the prophylaxis of infectious complications secondary to gastrointestinal and gynecologic surgery, and may have a role in surgical prophylaxis as it is well tolerated by most patients and is cost-effective. Indeed, the discovery of sulbactam provides us with yet another useful compound. Additional study is clearly needed so as to best use sulbactam to its fullest advantage. PMID- 3042859 TI - The influence of physical disease and its treatment on driving performance. PMID- 3042860 TI - The influence of psychiatric disease and its treatment on driving performance. PMID- 3042861 TI - Different distribution of ELI-perikarya in rat mesencephalon using various injection sites for colchicine. AB - In the present work, using the indirect peroxidase-labelled antibody technique, we made a comparative study about the distribution of Met-enkephalin-like immunoreactive (ELI)-cell bodies in rat mesencephalon; comparing animals that received colchicine into the lateral ventricle and animals that got the colchicine injected into different cerebral nuclei. We discuss the possibility that the local intracerebral injections are more effective than the intraventricular ones and we show that the results depend on the injection place. So, we think that there are possible met-enkephalinergic connections between the injected region and their corresponding immunoreactive mesencephalic nuclei. PMID- 3042862 TI - Homosexuality, suicide, and parasuicide in Australia. PMID- 3042863 TI - Homosexuality and mental health: a cross-cultural review. PMID- 3042865 TI - Special issues in the etiologies and treatments of sexual problems among gay men. AB - Some of the special circumstances and issues involved in both the causes and treatments of sexual problems among gay men are discussed. Reported frequencies of specific dysfunctions are discussed, and desire, arousal, and orgasmic phases are used to describe the special problems gay men have. These categories are also used to explore some of the probable contributing factors and specific treatment issues and suggestions that have been found useful. Since there is some indication that inhibited ejaculation is a somewhat frequent problem among gay men and has been more difficult than some dysfunctions to treat, emphasis has been placed on possible causes and treatments that have been helpful. A category of sexually inexperienced gay men is also discussed. PMID- 3042864 TI - Homosexuals in Eastern Europe: mental health and psychotherapy issues. PMID- 3042866 TI - Psychopathology, homosexuality, and homophobia. PMID- 3042867 TI - Theoretical considerations in psychotherapy with gay men and lesbians. AB - This paper examines certain issues relevant to psychotherapy with gay men and lesbians. The roles of general factors in psychotherapy in relation to homosexuality, including the theoretical orientation of the therapist, the nature of the presenting problem, the mode of therapy, and the personal characteristics of the therapist, are discussed. Special issues of concern to gay men and lesbians, specifically the question of pathology, the amount of information the therapist has about homosexuality, the sexual orientation of the therapist, and the unique problems of gay men and lesbians, are reviewed. Finally, an overview of the role of homophobia in psychotherapy is presented, and the importance of further exploration in this area is discussed. PMID- 3042868 TI - A simplified automated procedure for generation of human lymphokine-activated killer cells for use in clinical trials. AB - The administration of lymphokine-activated killer (LAK) cells along with interleukin-2 (IL-2) can mediate regression of tumors in selected patients. A closed automated system utilizing commercial blood cell separators has been developed for washing and Ficoll-Hypaque (FH) separation of lymphocytes, for lymphocyte culture in polyolefin bags, and for concentration of LAK cells out of culture prior to infusion. We now demonstrate that preparation of LAK cells can be simplified by elimination of the FH sedimentation step. Patient leukapheresis was performed using Fenwal CS-3000 blood cell separators, with a mean cellular yield per procedure of 54 X 10(9) WBC (95% lymphocytes), 184 X 10(9) RBC, and 306 X 10(9) platelets (n = 22). These cells were then washed in the same apheresis kit with a counter-current flow of saline, thereby eliminating 85% of platelets while retaining 88% of WBC. Aliquots of the washed cells were separated on FH gradients in 50 ml centrifuge tubes, and both FH-separated and washed-only cells were cultured at 3 X 10(6)/ml with 1500 U/ml IL-2 in polyolefin bags. Cytotoxicities of 22 preparations of LAK cells from 14 patients were evaluated in 4 h 51Cr release assays. Cells that were washed-only averaged 47, 35, and 9 lytic units/10(6) cells against K562, Daudi, and fresh tumor, while FH separated cells averaged 46, 33, and 6 lytic units/10(6) cells respectively. Cellular recoveries using the wash-only technique were 25% greater than when using FH sedimentation. Omission of FH separation saves time and expense in preparation and provides greater numbers of LAK cells for use in adoptive immunotherapy. PMID- 3042869 TI - Determination of lymphocyte subpopulations by enzyme immunoassay. Comparison with conventional fluorescence microscopy. AB - An enzyme immunoassay for measuring lymphocyte subpopulations was evaluated and the results compared with those obtained with conventional fluorescence microscopy, using two different panels of antibodies. The enzyme immunoassay is a photometric method which expresses the results using a standard curve with known amounts of cells. The method was reproducible and accurate. The intra-assay variation for the standard curve ranged from 3.2 to 5.7% and the interassay variation from 9.5 to 13.8%. The intra-assay variation for clinical samples ranged from 3.3 to 8.2%. Results obtained with the enzyme immunoassay and with conventional fluorescence microscopy showed a significant correlation (P less than 0.05) for all the subclasses of lymphocytes tested using two different panels of monoclonal antibodies. We conclude that the choice of method should be related to the particular needs and manpower in the laboratory. PMID- 3042870 TI - Time-resolved fluorescence using a europium chelate of 4,7-bis (chlorosulfophenyl)-1,10-phenanthroline-2,9-dicarboxylic acid (BCPDA). Labeling procedures and applications in immunoassays. AB - We describe optimal conditions for protein labeling with a new fluorescent probe, 4,7-bis-(chlorosulfophenyl)-1,10-phenanthroline-2,9-dicarboxylic acid (BCPDA). The labeled proteins are suitable for time-resolved fluorometric applications because BCPDA forms fluorescent complexes with Eu3+ which exhibit very long fluorescence lifetimes. Labeling parameters such as the organic solvent used, pH, protein concentration, BCPDA excess and incubation times, were optimized accordingly. Excess BCPDA was removed by gel filtration and labeled proteins were characterized by absorbance and fluorescence measurements. The effect of labeling on the biological (binding) activity of the proteins streptavidin, avidin, monoclonal and polyclonal antibodies was also studied. It is shown that the labeled antibodies can be used for time-resolved fluoroimmunoassay applications. PMID- 3042871 TI - Immunization with rough mutants of Salmonella minnesota: protective activity of IgM and IgG antibody to the R595 (Re chemotype) mutant. AB - We evaluated the immunoglobulin class responsible for the protective activity in serum obtained from humans and rabbits after immunization with the R595 (Re chemotype) mutant of Salmonella minnesota. Whole serum obtained before immunization and the IgG and IgM fractions failed to protect mice against lethal challenge with viable Klebsiella pneumoniae or Morganella morganii or with Salmonella typhi lipopolysaccharide (LPS). The protective activity of postimmunization serum resided solely in IgM antibody, whereas IgG antibody exhibited no protective activity. IgM antibody to the Re mutant was protective against bacterial challenge with both test strains of bacteria and S. typhi LPS. IgM antibody, at approximately the same concentration present in postimmunization serum, increased the LD50 of K. pneumoniae from less than 8.0 x 10(2) to greater than 2.0 x 10(4). These findings indicate that commercially prepared human IgG with high titers of antibody to antigens of the core portion of LPS would have little clinical utility. PMID- 3042872 TI - Immunization with rough mutants of Salmonella minnesota: initial studies in human subjects. AB - Vaccines prepared from unheated and boiled, acetone-precipitated Salmonella minnesota R595 (Re chemotype mutant) were administered subcutaneously to 122 healthy volunteers. Titers of antibody to Re lipopolysaccharide, the basal core structure of endotoxin, as measured by indirect hemagglutination, rose in a dose responsive fashion after immunization. Febrile reactions, usually mild, occurred after 7% of injections with the highest doses (2.0 and 3.0 x 10(10) organisms), and mild local soreness and tenderness were noted after approximately one-third of injections. Passive immunization of mice with sera from immunized subjects demonstrated that protective activity against challenge with both heterologous, viable gram-negative bacilli and endotoxin developed. Although measuring serum protective activity, developing a serological assay that correlates with protective activity, and potential vaccine toxicity remain problems, immunization of humans with the Re mutant results in serum protective activity against endotoxin and viable bacilli and has the potential for clinical usefulness. PMID- 3042873 TI - Steroids are out in the treatment of cerebral malaria: what's next? PMID- 3042874 TI - High-dose dexamethasone in quinine-treated patients with cerebral malaria: a double-blind, placebo-controlled trial. AB - We compared placebo and dexamethasone (initial dose, 3 mg/kg; total, 11.4 mg/kg per 48 h) in a double-blind trial involving 10 stuporous and 28 comatose patients with cerebral malaria. Patients were 18 mo to 42 y of age (geometric mean, 10.2 y), and the 19 patients in each group were comparable on admission. All patients received intravenous quinine therapy. Four patients (21%) in each group died. There were no significant differences between the placebo- and dexamethasone treated groups in time until patients became afebrile (median, 51 vs. 19 h), the level of consciousness became normal (mean, 80 vs. 83 h), or parasitemia was cleared (mean, 2.1 vs. 3.4 d) or in the incidence of complications. Coma or hyperparasitemia (greater than or equal to 5% of erythrocytes parasitized) at the time of admission and hypoglycemia at any time during hospitalization were significantly correlated with a fatal outcome, which was not improved by using dexamethasone. We conclude that high-dose dexamethasone is not indicated for treating cerebral malaria. PMID- 3042876 TI - Cross-protection by B subunit-whole cell cholera vaccine against diarrhea associated with heat-labile toxin-producing enterotoxigenic Escherichia coli: results of a large-scale field trial. AB - The B subunit (BS) of cholera toxin and that of the heat-labile enterotoxin (LT) of enterotoxigenic Escherichia coli (ETEC) are antigenically similar. We therefore assessed whether a combined cholera toxin BS/whole-cell (BS-WC) oral vaccine against cholera conferred cross-protection against LT-producing ETEC (LT ETEC) diarrhea in a randomized, double-blind field trial among rural Bangladeshi children and women. The 24,770 persons who ingested two or more doses of BS-WC vaccine were compared with 24,842 controls who took two or more doses of killed whole-cell (WC) oral cholera vaccine. Sixty-seven percent fewer episodes of LT ETEC diarrhea were noted in the BS-WC group than in the WC group during short term (three-month) follow-up (P less than .01), but no reduction was evident during the ensuing nine months. Short-term protection was particularly notable against LT-ETEC diarrhea causing life-threatening dehydration (protective efficacy, 86%; P less than .05). PMID- 3042875 TI - Comparative pharmacokinetics and pharmacodynamics of amikacin and ceftazidime in tricuspid and aortic vegetations in experimental Pseudomonas endocarditis. AB - A factor in the higher medical cure rates for endocarditis in the right as opposed to the left side of the heart in humans may be a difference in antimicrobial pharmacokinetics within vegetations. Rabbits with combined tricuspid and aortic endocarditis due to Pseudomonas aeruginosa received single intravenous doses of either ceftazidime (50 mg/kg) or amikacin (15 or 40 mg/kg). For each antibiotic regimen, areas under the time-concentration curves and percent vegetation penetrances were significantly greater for tricuspid than aortic vegetations (P less than .001). Time-concentration curves for aortic vegetations paralleled those for plasma; curves for the tricuspid vegetations resembled those for subcutaneous fibrin clots. The times above the minimum bactericidal concentration for tricuspid vegetations were significantly longer than those achieved within aortic vegetations for ceftazidime (P less than .01) and amikacin at 15 mg/kg (P less than .001). Antimicrobial pharmacokinetics and pharmacodynamics may be more favorable within tricuspid than aortic vegetations; this difference may, in part, explain more salutary outcomes in bacterial endocarditis involving the right side of the heart. PMID- 3042877 TI - Lectin-like attachment sites on murine pulmonary alveolar macrophages bind Aspergillus fumigatus conidia. AB - Murine pulmonary alveolar macrophages bound Aspergillus fumigatus conidia in vitro at 4 C and 37 C in the absence of serum or opsonins. This attachment was dependent on calcium and was sensitive to mild trypsinization and paraformaldehyde pretreatment of the macrophage membrane. Chitotriose, N acetylglucosamine, D-mannose, alpha-methyl-mannoside, and L-fucose, but not D galactose, were effective inhibitors of conidial binding. This pattern of reduction of conidial binding was consistent with that for the mannosylfucosyl receptor. In addition, conidial binding may be mediated by another lectin on the macrophage membrane, one that recognizes chitin components, because N acetylglucosamine and chitotriose exhibited greater inhibition than expected for the mannosyl-fucosyl lectin. PMID- 3042878 TI - Effect of immunosuppression and amphotericin B on Aspergillus antigenemia in an experimental model. AB - A modified enzyme-linked immunosorbent assay (ELISA) for crude carbohydrate antigen was used to evaluate the kinetics of aspergillus antigenemia and to determine the effect of therapy on circulating antigen levels in an experimental model. The ELISA was rapid, simple to perform, and able to detect less than 10 ng of antigen/mL of serum. The model was also used to evaluate the effect of temporary and persistent immunosuppression on experimental disease. Antigen levels rose progressively in untreated control rabbits; all 15 animals had significant antigenemia. Treated animals had markedly reduced antigen levels, but nine of 13 rabbits had detectable antigen after 72 h of therapy, a result that correlated with persistent disease. Therapy begun 24 h after challenge in temporarily immunosuppressed animals was more likely to sterilize tissues than was therapy begun 48 h after challenge. Therapy in persistently immunosuppressed rabbits was less effective and may require improved antifungal regimens to be successful. PMID- 3042881 TI - Penicillin tolerance in group A streptococci. PMID- 3042879 TI - Acute measles in patients with and without neurological involvement: distribution of measles virus antigen and RNA. AB - Using peroxidase immunohistochemistry and in situ hybridization to localize viral antigen and RNA, we studied autopsy tissues from 20 cases of acute fatal human measles (including seven patients with acute encephalomyelitis) and peripheral blood mononuclear cells from 16 patients with acute, nonfatal measles. In immunologically normal patients, virus was detected in five of nine who died five days or less after the onset of rash but in none of 11 who died later. Virus was localized to epithelial cells of lung, gut, bile duct, bladder, and skin and to lymphoid organs. Neither viral antigen nor RNA was detected in brain sections from 14 patients, including seven with acute encephalomyelitis and four with virus identified in other tissues, a finding supporting an indirect pathogenesis of post-measles encephalomyelitis. These data show that measles virus replicates in cells previously not recognized to be involved (capillary endothelium of lymph node and thymus, Hassall's corpuscles, and hepatic duct epithelium) and that invasion of the brain parenchyma during acute measles is uncommon. PMID- 3042882 TI - Human melioidosis and biologic false-positive reactions in unrelated serological tests. PMID- 3042880 TI - Fibronectin and age-limited susceptibility to type III, group B Streptococcus. PMID- 3042883 TI - Q fever in patients with hepatitis and pneumonia: results of laboratory-based surveillance in the United States. PMID- 3042884 TI - Prophylactically administered Retrovir in health care workers potentially exposed to the human immunodeficiency virus. PMID- 3042885 TI - Fibrous component of yeast mitochondria. AB - An intramitochondrial fibrous component (IMF) was consistently detected by electron microscopy in all eight strains of yeast examined, either respiratory competent or deficient, and including two species, Saccharomyces cerevisiae and S. uvarum. IMF was always found assembled into a layer 20 nm in thickness. In respiratory-deficient mutants, multiple IMF layers roll up concentrically to give rise to cylindrical inclusion bodies which attain several micrometers in length. In normal respiring yeasts, IMF occurs in close association with cristae to elaborate composite structures in which multiple parallel IMF layers are sandwiched between a pair of cristae. Evidence is presented to demonstrate the extramitochondrial origin of IMF. PMID- 3042886 TI - Structure and location of tellurium deposited in Escherichia coli cells harbouring tellurite resistance plasmids. AB - The plasmids RP4Ter and pHH1508a, which belong to the P and HII incompatibility groups, respectively, confer resistance to potassium tellurite (K2TeO3) on Escherichia coli. The genes for tellurite resistance were cloned from each plasmid onto the vector pUC8 to create pDT1366 and pD1364, respectively. Unstained, unfixed bacteria carrying these plasmids contained black intracellular deposits when grown on media containing tellurite. Thin sections of these bacteria fixed with glutaraldehyde were prepared and examined by electron microscopy. The black deposits were located inside the cell and were frequently associated with the inner membrane of the bacterium. Bacteria containing pDT1366 or pDT1364, and therefore a higher gene dosage of the Ter determinant, contained more black deposits, but had a decreased resistance, as measured by the minimum inhibitory concentration using the agar dilution method. Using the technique of electron spectroscopic imaging, the black intracellular deposits were shown to contain predominantly reduced metallic tellurium, and significant amounts of oxygen or carbon, thereby confirming earlier results using X-ray diffraction analysis of whole cells. PMID- 3042887 TI - Electron spectroscopic imaging of DNA. AB - Electron spectroscopic imaging (ESI) was carried out using a fixed beam electron microscope equipped with a parallel electron energy filter to form micrographs of purified plasmid DNA without the use of heavy metal stains and shadows. Inelastically scattered electrons that have ionized the phosphorus LII,III shell electrons were used to form phosphorus distribution maps of DNA and deoxyribonucleoprotein complexes. Signal-to-noise values of the net phosphorus content over single DNA molecules compared to two and four interwound DNA strands directly reflect the known stoichiometric levels of phosphorus content, illustrating that ESI can be used to determine the relative levels of nucleic acid in nucleoprotein complexes. An initial attempt to characterize nucleosomal and transcriptionally active chromatin from Saccharomyces cerevisiae with this technique reveals three distinct ultrastructural classes of the basic chromatin fiber. PMID- 3042888 TI - [Structures of pyruvate kinase isozyme genes and regulation of their expressions]. PMID- 3042890 TI - [Vitamin C and cell growth: use of L-ascorbic acid 2-phosphate in culture for construction of three-dimensional structure from isolated cells]. PMID- 3042889 TI - [Molecular and cellular biology of lymphokines produced by antigen activated T cells]. PMID- 3042891 TI - [Intercellular fate of nitrate reductase in higher plants--degradation and inactivation mechanisms]. PMID- 3042892 TI - [Structure and function of lipopolysaccharides from genus Rickettsia]. PMID- 3042893 TI - [Transfection of mitochondrial genomes of higher animals]. PMID- 3042894 TI - [Recent view on the last step of the biosynthetic pathway of tetrahydrobiopterin]. PMID- 3042895 TI - [Pectinases and their genes of genus Erwinia]. PMID- 3042896 TI - [A case of hypoglycemic coma associated with myoclonus, periodic synchronous discharges and progressive cerebral atrophy resembling Creutzfeld-Jakob disease]. PMID- 3042897 TI - [A case of mineralocorticoid-resistant renal hyperkalemia without sodium wasting (type II pseudohypoaldosteronism)]. PMID- 3042899 TI - [A case of marfanoid body habitus associated with an excessive hyperextensibility of the skin--an unclassified case in inherited connective tissue diseases]. PMID- 3042898 TI - [A case of Liddle's syndrome with familial occurrence]. PMID- 3042900 TI - [A case of migraine with transient monocular blindness, hemiparesis and 6 Hz positive spikes on electroencephalogram]. PMID- 3042901 TI - [A case of malignant thymoma associated with cardiac tamponade]. PMID- 3042903 TI - [Histopathological study on pulpal response to restorative composite resins and their ingredients]. PMID- 3042902 TI - [Some problems on dental alloys]. PMID- 3042904 TI - Comparison of catalytic activity and mass concentration of serum creatine kinase MB isoenzyme in the detection of coronary reperfusion in acute myocardial infarction after therapeutic thrombolysis. AB - Serum creatinine kinase MB isoenzyme time-activity curves are useful for the assessment of coronary reperfusion after acute myocardial infarction. The purpose of this study was to compare serum creatine kinase MB catalytic activity with mass concentration for the determination of coronary reflow after therapeutic thrombolysis. Creatine kinase MB mass was determined immunoenzymometrically. Creatinine kinase MB catalytic activity concentration was determined by electrophoresis. Serum was collected every 4 hours for 96 hours in two groups of myocardial infarction patients: A (n = 10), urokinase induced reperfusion; B (n = 10), conventional therapy without urokinase. Peaks of mass and activity occurred at similar times in groups A and B. Both were significantly earlier in the urokinase treated patients. The maximal rate of increase of creatine kinase MB (based on either mass or catalytic activity) was threefold greater in the urokinase group. There are no important differences between the behaviour of creatine kinase measured as catalytic concentration or as mass concentration. Mass concentration is therefore equally useful as an indicator of coronary reperfusion. PMID- 3042905 TI - Assessing obstetric risk. A review of obstetric risk-scoring systems. AB - The primary purpose of formal risk assessment in obstetrics is the prevention and consequent reduction of perinatal morbidity and mortality through early identification and intervention. Obstetric risk scoring quantifies identified risk factors according to their relative contribution to adverse perinatal outcomes and aggregates individual factor scores. A review of existing scoring methods reveals consistently low positive predictive values and more accurate prediction when the assessment occurs closer to the time of actual delivery. While numerous scoring systems exist in the literature, few are convenient in practice, and none appear to assess effectively the dynamic character of pregnancy. PMID- 3042907 TI - The adequacy of treating depression. PMID- 3042906 TI - Bacterial vaginosis. AB - Bacterial vaginosis (nonspecific vaginitis) is a polymicrobial, superficial vaginal infection caused by an increase in anaerobic organisms and a concomitant decrease in lactobacilli. Gardnerella vaginalis, once thought to be the sole etiologic agent, is probably one of several endogenous members of the vaginal flora that overgrow in women with bacterial vaginosis. Whether the growth of anaerobes or a primary decrease in lactobacilli is the initial pathogenic event remains unclear. Epidemiological studies have revealed that current or previous infections caused by Trichomonas organisms, increased sexual activity, and intrauterine device use are risk factors for this condition. Studies have indicated that bacterial vaginosis, previously thought to be a benign illness, is associated with some morbidity in pregnant women. Symptoms remain unreliable in the diagnosis of bacterial vaginosis. Diagnostic efficacy is best achieved by utilizing clinical signs. Assessment of cure is best accomplished by Gram stain, not clinical criteria. Metronidazole, 500 mg orally for seven days, remains the treatment of choice; however, a 2-g single dose of metronidazole represents a reasonable alternative if cost and compliance issues predominate in a clinical situation. Although a recent study supports the contention that treatment of the male sexual partner of women with bacterial vaginosis is effective, a general recommendation cannot be made with confidence on the issue of sexual partner treatment until other supporting work is done. PMID- 3042908 TI - The case for and concerns about continuous dopamine stimulation in Parkinson's disease. AB - Delivery of levodopa or other dopaminergic agents to the striatum is a critical determinant of the clinical response and may account for many of the fluctations in response ("on-off"). This should be manageable with innovative delivery systems for the drugs. However further studies are required to determine if continuous dopaminergic stimulation will be complicated by increased levels of drug metabolites or down regulation of dopaminergic neurotransmission. PMID- 3042909 TI - Psychiatric side effects during the treatment of Parkinson's disease. AB - Dopaminergic agents including both levodopa and direct-acting agonists induce a variety psychiatric side-effects of which psychosis is the most significant. When this occurs early in the course of treatment, there is usually a history of prior psychotic illness. Chronic treatment can, however, elicit psychosis in individuals without such a history. The possible pathogenesis of this is reviewed. PMID- 3042910 TI - Applications of new drug delivery technologies to Parkinson's disease and dopaminergic agents. AB - Recent advances in drug delivery technology are creating novel therapeutic approaches to the treatment of Parkinson's disease with levodopa and dopamine agonists. This article reviews those technologies which can be applied to Parkinson's disease, both for targetting the central nervous system with drugs, as well as for matching the appropriate rate controlled delivery with therapeutic needs. In particular, the possibility exists for eliminating erratic highs and lows of drug delivery to the brain, and to substitute rate controlled, constant drug delivery. Clinical investigations now in progress suggest that new technologies which deliver constant dopaminergic stimulation to patients with Parkinson's disease may not only eliminate the unpredictable swings in therapeutic efficacy in Parkinson patients with the "on/off" effect, but may even have a role in the future in preventing such fluctuations from developing in patients chronically treated with dopaminergic therapies. PMID- 3042912 TI - Pathogenetic studies of motor fluctuations in Parkinson's disease. AB - Pharmacokinetic and pharmacodynamic mechanisms for levodopa have been studied in relation to the pathogenesis of the motor fluctuations which complicate advanced Parkinson's disease. Since levodopa clearance from the general circulation was found to be similar in patients with wearing-off or on-off phenomena and those with a stable response to levodopa, peripheral pharmacokinetic factors are unlikely to be involved. Efficacy half-time for levodopa, on the other hand, was significantly reduced in patients with mainly wearing-off fluctuations in comparison to those manifesting a stable response to oral levodopa; individuals with predominantly on-off phenomenon had an even more extreme reduction in the duration of the antiparkinsonian action of levodopa. Conversion from oral to intravenous levodopa treatment immediately stabilized plasma levodopa levels in both the wearing-off and on-off groups; motor variability also decreased, especially in those with wearing-off phenomenon. During 11 days of continuous intravenous levodopa therapy, additional reductions in motor fluctuations occurred in both groups, but at a significantly faster rate in patients with wearing-off than in those with on-off responses. These results suggest that wearing-off phenomenon may arise as a consequence of the degeneration of dopamine terminals due to natural disease progression with a resultant inability to buffer variations in levodopa availability; on-off phenomenon, may reflect additional postsynaptic dopamine receptor dysregulation, possibly in response to the resultant, nonphysiologic fluctuations in synaptic dopamine. PMID- 3042911 TI - Continuous dopaminergic stimulation: state of the art and outlook. PMID- 3042913 TI - A combined regimen of subcutaneous lisuride and oral Madopar HBS in Parkinson's disease. AB - At the first stage of a pilot study involving 14 parkinsonians with motor fluctuations, treatment with standard Madopar was substituted by a sustained release form, Madopar HBS, which attenuated fluctuations in patients with end-of dose impairment, but achieved only moderate improvement in patients with on-off phenomena. In a second phase of the trial, 4 parkinsonians exhibiting the most severe fluctuations of mobility and the poorest response to Madopar HBS (Hydrodynamically Balanced System) were selected for treatment with a combined regimen utilizing subcutaneous lisuride infusions as the additional component. The sequence of the trial was as follows: 1. standard Madopar, 2. Madopar HBS, 3. standard Madopar combined with lisuride infusions and 4. Madopar HBS combined with lisuride infusions. Steady improvement was observed along the lines of this schedule, but the best results were obtained when Madopar HBS was combined with lisuride infusions. Subsequently motor fluctuations were less marked or disappeared, early-morning Parkinson symptoms decreased and dystonia was not recorded any longer. Even better results could be accomplished in an extended trial attempting to establish the best dosage ratio of the combination, possibly admitting increased dosage. The tolerance of the combined regimen was excellent, except in one patient who transiently exhibited delusions and postural hypotension. The combination of sustained-release Madopar and continuous infusions of the dopaminergic agonist lisuride seems to prove a more physiological and effective regimen for the treatment of severe motor fluctuations. PMID- 3042915 TI - The thalamic syndrome of Dejerine and Roussy. PMID- 3042914 TI - Brown-Sequard's description of spontaneous cerebellar haemorrhage. PMID- 3042916 TI - Stabilisation of the infected spine. AB - A metal stabilising rod was inserted at the time of surgical drainage of the abscess in six patients with spinal instability secondary to vertebral osteomyelitis. The procedure enabled early postoperative mobilisation to be undertaken in five patients. All cases obtained immediate relief of pain. One case of delayed wound healing occurred but not late recurrent infections were encountered after discontinuing antibiotic therapy. The technique was employed in cases of pyogenic as well as tuberculous spinal osteomyelitis. PMID- 3042917 TI - St Paul and temporal lobe epilepsy. PMID- 3042919 TI - Schwannoma of the medulla oblongata. Case report. AB - A case of intraparenchymal schwannoma of the medulla oblongata is presented. The radiographic and pathological characteristics of this rare tumor are discussed, and the world literature regarding intracerebral intraparenchymal schwannomas is reviewed. The current etiological theories of these intra-axial nerve-sheath tumors are reviewed. The importance of early identification and differentiation of these potentially curable tumors from malignant glial tumors is emphasized. PMID- 3042918 TI - Corticotectal circuit in the cat: a functional analysis of the lateral geniculate nucleus layers of origin. AB - 1. The dorsal lateral geniculate nucleus (LGN) of the cat is a major thalamic relay between the retina and several visual cortical areas. These cortical areas in turn project to the superior colliculus (SC). The aim of the present experiment was to determine which LGN layers provide a necessary input to the corticotectal circuit. 2. Individual layers of the LGN were reversibly inactivated by microinjection of cobalt chloride during recording of visual responses in the retinotopically corresponding part of the superior colliculus. 3. For cells driven through the contralateral eye, inactivation of layer A or the medial interlaminar nucleus (MIN) had little effect on visual responsiveness in the superior colliculus. In contrast, inactivation of layer C abolished visual responses at one-quarter of the SC recording sites, reduced responses at another quarter, and left half of the recording sites unaffected. 4. For cells driven through the ipsilateral eye, inactivation of layer C1 or the MIN had no effect. Inactivation of layer A1 uniformly reduced visual responses in the superior colliculus and usually abolished them entirely. 5. These results are compatible with previous work showing that cortical input to the SC originates from Y-cells. They indicate that two of the five Y-cell containing layers (A1 and C) provide major inputs to the corticotectal circuit. The results suggest that layer A1 is functionally allied to layer C as well as to layer A. PMID- 3042921 TI - Limiting artifact in CT stereotaxic periventricular procedures. Technical note. AB - One potential source of computerized tomography image artifact during stereotaxic data acquisition is scatter from standard steel-tipped fixation pins as supplied for use with the Brown-Roberts-Wells stereotaxic headframe. In some cases, this can hinder selection of periventricular targets. Replacement of the steel-tipped pins with aluminum-tipped pins, which produce less artifact, helps to facilitate stereotaxic intervention. PMID- 3042920 TI - Infection-related spontaneous atlantoaxial dislocation in an adult. Case report. AB - This paper reports the third described case of infection-related atlantoaxial subluxation in an adult. Like most of the similar cases seen in the pediatric literature, this case was associated with a parapharyngeal beta-hemolytic streptococcal abscess. Based upon this experience, the authors advocate intravenous antibiotic therapy and 1) immediate reduction followed by application of a halo brace; 2) immobilization in a halo brace for at least 3 months; and 3) a C1-2 wiring and fusion procedure for patients who fail this trial of conservative therapy. PMID- 3042922 TI - Nimodipine therapy in poor-grade aneurysm patients. PMID- 3042923 TI - Renal scans in pregnant transplant patients. AB - This study demonstrates the normal technetium-99m diethylenetriaminepentaacetic acid ([99mTc]DTPA) renal scan in pregnant patients with transplanted kidneys. Five pregnant renal transplant patients had seven [99mTc]DTPA renal studies to assess allograft perfusion and function. All scans showed the uteroplacental complex. The bladder was always compressed and distorted. The transplanted kidney was frequently rotated to a more vertical position. In all patients allograft flow and function were maintained. There was calyceal retention on all studies and ureteral retention activity in three of five patients. Using the MIRD formalism, the total radiation absorbed dose to the fetus was calculated to be 271 mrad. This radiation exposure is well within NRCP limits for the fetus of radiation workers and an acceptable low risk in the management of these high risk obstetric patients. PMID- 3042924 TI - Semiquantitative in vitro binding assay of indium-111-labeled monoclonal antibodies to human cancer and normal tissues. AB - We have developed a simple in vitro method for the semiquantitative assessment of the radiolabeled antibody binding to cancer and normal tissues. Indium-111 labeled F(ab')2 fragments of 17-1A and 19-9 monoclonal antibodies with well characterized specificity for gastrointestinal cancer demonstrated similar binding properties between cultured cancer cells and membrane fractions of homogenates prepared from tumor tissues. All of the 17 colon cancer specimens and seven (64%) of 11 gastric cancer specimens obtained by surgery showed positive binding with 17-1A. Specific binding of 19-9 was observed in 9 (53%) colon cancers and 4 (36%) gastric cancers. However, some normal colon tissues were also positive with 111In-labeled 17-1A. Relative levels of CA 19-9 antigen expression, determined by the binding with radiolabeled antibodies, correlated with percent positive cells determined by the immunohistochemical assays. Furthermore, membrane fractions could be cryopreserved without losing antibody-binding activity. These results indicate that this assay can be used for testing the immunoreactivity of radiolabeled anti-tumor antibodies and in vitro binding properties to cancer and normal tissues. PMID- 3042925 TI - The expanding role of MIBG in clinical medicine. PMID- 3042926 TI - Reduced saphenous vein prostacyclin production in the absence of endothelial detachment: a marker of functional damage by high potassium cardioplegic solutions to saphenous vein bypass grafts. PMID- 3042927 TI - Developing trauma care systems: the trauma nurse coordinator. AB - Emergency medical services/trauma care systems offer unique opportunities for nurse administrators in community hospitals and medical centers to encourage a professional practice model for trauma care in the department of nursing. The first article in this series (May 1988) provided an overview of trauma care and the planning and operational functions of nursing departments in trauma care facilities. In this article, the authors acknowledge trauma nursing as a specialty and discuss the role and functions of the trauma nurse coordinator in organizing and providing trauma care. PMID- 3042928 TI - JONA's semiannual directory of consultants to nursing administration. PMID- 3042929 TI - An overview of sexually transmissible diseases in the perinatal period. PMID- 3042930 TI - Prolonged pregnancy and the biophysical profile. A birthing center perspective. PMID- 3042931 TI - The cafeteria diet as a tool for studies of thermogenesis. AB - The cafeteria diet involves feeding experimental animals a choice of palatable human food items to stimulate energy intake, and has been used extensively to study diet-induced thermogenesis. In a recent commentary Moore has argued that this feeding regime is inappropriate for such studies because the nutrient composition cannot be controlled, many of the effects seen are due to protein or nutrient deficiency and accurate measurements of energy intake are difficult to achieve. We argue that all of these criticisms can be overcome by careful use of the feeding regime and well-controlled experiments. Gross nutrient composition of cafeteria diets can be modified over a wide range, and such studies demonstrate that the effects of protein deficiency can be clearly dissociated from those of hyperphagia. There is no experimental evidence for nutritional deficiency in cafeteria-fed animals even over very long periods of time. Furthermore, the alternatives suggested by Moore, i.e., presenting sucrose solutions to drink or high fat diets, suffer the same drawbacks of altered and often uncontrolled nutrient intake and yet produce little or no increase in energy intake. Criticism of the cafeteria diet is not justified simply because of its misuse by nutritionally naive experimenters. The value and validity of this feeding regime is further supported by the enormous impact it has made on our understanding of energy balance regulation and thermogenesis. PMID- 3042934 TI - [Bleeding of sinusectomy for chronic sinusitis]. PMID- 3042932 TI - Planning with elderly outpatients for contingencies of severe illness: a survey and clinical trial. AB - The authors examined whether elderly patients would report positive or adverse emotional effects after their doctor, during a routine clinic visit, asked them to begin planning for future serious illness. Seventy-four patients, 65 years old or older, who were followed at a university hospital medical clinic were randomly allocated to an intervention or a control group. The intervention was a detailed discussion with the patient's physician of the patient's wishes about decision making and life support therapy in the event of extreme or incapacitating illness. A blinded interviewer then asked all consenting patients how they felt about the physician, the clinic visit, and their medical care. Intervention-group patients were questioned about their reactions to the physician and the discussion. Four important findings emerged: 1) Some emotional uncertainty was created when doctors raised these questions unexpectedly: one patient became visibly upset during the discussion, and three who gave consent to be interviewed afterward said that the discussion had made them wonder about their health. Nonetheless, all patients who received the intervention and completed the study were pleased that their doctor had asked. 2) Only 44% of all consenting patients reported having discussed these issues previously; only one had done so with a doctor. 3) 97% of patients who responded wanted to be kept informed by the doctor about their medical situations in times of serious illness. 4) Patients' replies to specific questions about life-sustaining therapy in the event of their own severe illnesses were quite variable. During routine clinic visits doctors can encourage most elderly patients to begin specific planning for potential severe illnesses. PMID- 3042936 TI - Digestive and absorptive phase anomalies associated with the exocrine pancreatic insufficiency of cystic fibrosis. AB - The pancreas has an enormous reserve capacity, and significant malabsorption usually signals complete absence of exocrine function. However, there is evidence that acid lipases of nonpancreatic origin play an important compensatory role. Complete duodenal hydrolysis of fat requires a series of complex interdependent physicochemical events involving pancreatic lipase, colipase, phospholipase A2, and bile salts in an environment where the pH must be close to neutrality. Lipolytic products must then be shuttled through the unstirred water layer to the surface of the microvillus membrane by ionized bile salts, which must be present in sufficient concentrations to form micelles. In pancreatic insufficiency, there is not only a defective lipolytic phase but also an impaired micellar phase. The output of bile salts is decreased because of increased fecal loss. Furthermore, a significant percentage of bile salts precipitate because the duodenum is acidic and there is a large predominance of glycine conjugates. Although much less work has been done on the absorptive phase of patients with pancreatic insufficiency, there is tentative evidence that defective phospholipid absorption, essential fatty acid deficiency, and protein malnutrition could impair the absorptive phase, particularly chylomicron formation. Although significant advances have been made in our understanding of factors responsible for malabsorption associated with pancreatic insufficiency, much remains to be done for the further delineation of defects. It is hoped that this will lead to further refinements of enzyme preparations and to new strategies of intervention. PMID- 3042935 TI - [A case of progressive bilateral hearing loss due to Paget's disease]. PMID- 3042933 TI - The severity model of chronic headache. PMID- 3042937 TI - The impact of nutrition in cystic fibrosis: a review. AB - A high energy intake, compensating for malabsorption, and the energy cost of lung disease, lung infections, and the underlying metabolic abnormality, is required to ensure normal growth in patients with cystic fibrosis. This goal can be readily achieved by adherence to a high quantity, normally balanced diet (with 40% of the energy as triglycerides of long-chain fatty acids). In contrast, this goal cannot be reached in the majority of patients adhering to low-fat and thus low-energy-containing diets, which almost inevitably lead to malnutrition with wasting and stunting. The prevention of malnutrition may well have considerably enhanced the prognosis of patients at one clinic. Further work is needed to define the interrelationship of nutrition and lung disease, and to define the appropriate nutrient requirements induced by the lung disease per se, recurrent infections, and the underlying disease process. PMID- 3042938 TI - Efficacy of pancreatic enzyme supplementation in children with cystic fibrosis: comparison of two preparations by random crossover study and a retrospective study of the same patients at two different ages. AB - Two studies were performed in children with cystic fibrosis (CF) to compare the efficacy and tolerability of pancreatic enzymes prepared as enteric-coated microspheres with that of a conventional enzyme preparation. The parameters evaluated included the following: fecal fat excretion, coefficient of fat absorption, daily caloric intake, percent of diet as fats, proteins and carbohydrates, increase in height and weight, frequency and consistency of stools, palatability of the preparation, and patient compliance. The first study was an open randomized crossover trial of an enteric-coated microsphere preparation (Pancrease) versus conventional pancreatin given alone or with cimetidine. With Pancrease compared to conventional pancreatin, a significant improvement was observed in all the digestive parameters in addition to better patient compliance. In the second study, the response of a group of CF patients given Pancrease for at least 3 months (3-67 months) was retrospectively compared with the response to conventional pancreatin, which had been given to the same patients from 10 months to 8 years earlier. In comparison to conventional pancreatin, Pancrease provided better digestive efficacy and greater increases in the growth rate of teenage patients. With Pancrease, the number of daily dosage units is decreased even when fat intake is increased. No adverse reactions were seen with either of the enzyme preparations used in these studies. PMID- 3042939 TI - Efficacy of pancrease: crossover comparative study versus eurobiol in cystic fibrosis. AB - An open crossover study in patients with documented cystic fibrosis (CF) compared the efficacy and acceptability of Pancrease (Cilag, PC), consisting of pH sensitive, enteric-coated microspheres, with those of Eurobiol, consisting of lyophilized total pancreas. Eleven patients, age 6-17 years, were hospitalized for study. A hypercaloric, normal fat diet as well as previous antibiotic, vitamin, and other CF therapy were maintained during the 2-week study period. During the first study week, eight patients were given four divided doses of either Pancrease, 8 capsules daily, and two patients were given Eurobiol, 2 bottles daily. One patient received six Pancrease capsules or 1.5 bottles of Eurobiol daily. Patients were switched to the alternate drug for the second study week. Stools were collected for analysis over the final 3 days of each week. Steatorrhea (mean +/- 1 SD) with Pancrease was 11.8 +/- 6.6 g/24 h and with Eurobiol was 17.9 +/- 11.7 g/24 h. The coefficient of lipid absorption was 74.1 +/- 13.3% for Pancrease and 58.6 +/- 22.5% for Eurobiol (p less than 0.02). The decrease in steatorrhea and increase in lipid absorption with Pancrease compared to Eurobiol were 33.9% and 26.5%, respectively. All patients considered the acceptability of Pancrease superior to that of Eurobiol. No adverse effects were seen with either drug. PMID- 3042940 TI - Nutritional management and pancreatic enzyme therapy in cystic fibrosis patients: state of the art in 1987 and projections into the future. PMID- 3042941 TI - Nutritional requirements in cystic fibrosis: a review. AB - Food intake is often low in cystic fibrosis (CF), although the patient usually needs more than the standard recommended daily allowance (RDA). Clinics giving food supplementation from an early age report improved survival and nutritional status. Nutritional improvement has been facilitated by improved forms of pancreatin. An additional calorie intake in CF is required to compensate for losses due to malabsorption and to allow for catch-up growth when necessary. With advanced pulmonary disease there are additional requirements for infection and increased work of breathing. There is also evidence for an increased basal metabolic activity in CF, perhaps related to the fundamental intracellular biochemical disorder. Together these factors add to a daily need for 120%-150% RDA for optimum growth and homeostasis. PMID- 3042942 TI - Historic landmarks of perinatal medicine in obstetrics. PMID- 3042943 TI - Longitudinal follow-up of 100 patients at risk of intrauterine growth retardation: comparison of diagnosis in two periods. AB - The concept of low birth weight includes two different entities: prematurity and intrauterine growth retardation. Both of them are major public health problems, because they increase perinatal morbidity and mortality Early diagnosis of IUGR leads to adequate decisions, making possible a reduction in perinatal morbidity and mortality. In order to make an early diagnosis of IUGR, clinical methods have proven to be insufficient. Ultrasonography is an important aid to this diagnosis, introducing the measurement of fetal diameters and perimeters. This study, designed to compare both methods, was carried out in at the Antoine Beclere Hospital, Clamart, France. Data processing was done in CLAP-PAHO/WHO. In the first period, retrospective analyses of 116 clinical histories with IUGR were performed. In this study, only fetal diameters were used and the accuracy of clinical and ultrasonographic diagnosis was evaluated. In the second period a prospective longitudinal follow-up study of 100 pregnant women at risk of developing IUGR was carried out. Clinical diagnosis was also evaluated, and compared to the ultrasonographic approach. The parameters used were the fetal diameters and perimeters (head and abdominal perimeters, and their relationship). Sensitivity, specificity and predictive values of the ultrasonic parameters were calculated (table I). The clinical and ultrasonographic diagnosis of both periods were compared with the purpose to analyze the effect of the measurement of fetal perimeters in the diagnostic accuracy. Newborns of the 100 patients in the prospective study were classified into two groups according to birth weight.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3042944 TI - Prenatal ultrasonographic findings in extralobar subdiaphragmatic lung sequestration--a case report. AB - We present the prenatal ultrasonographic findings of an echo-dense solid mass above the normal left kidney and below the diaphragm. This mass was diagnosed as extralobar lung sequestration (ELS) at laparotomy of the child. ELS (or accessory lungs) is a rare congenital abnormality defined as a lung segment outside a normal lung, usually localized in the left lower thorax. A subdiaphragmatic localization is rare. The prognosis depends upon both the presence (as found in over 50% of reported cases) and severity of additional congenital anomalies. PMID- 3042945 TI - Roberts syndrome: antenatal ultrasound--a case report. AB - There is obvious confusion and overlap involving these various syndromes, i.e., Roberts syndrome, SC phocomelia, and TAR syndrome. However, the patient reported here seemed to exhibit the features of shortened upper extremities, clefting, and microbrachycephaly characteristic of Roberts syndrome. Antenatal detection of Roberts syndrome may be important in that early neonatal death can be expected. Other similar syndromes may have much longer survival. Cytogenetic abnormalities may be detected as well. It appears that routine ultrasonographic assessment of humerus length in addition to femur length may yield certain diagnoses that otherwise would be missed. The parents can be counselled concerning antenatal management including the possibility of not performing a cesarean section for some abnormal presentations. PMID- 3042946 TI - Remembering your parents: reflections on the retrospective method. AB - In the present paper, I critique the use of the retrospective method when it is used as a proxy for actual longitudinal data on personality development. Studies on the constructive nature of memory cast strong doubts about the meaning of retrospective data. There are good reasons, both theoretical and empirical, to distrust the accuracy of such recall concerning parenting, whether recalled by parents, children, or siblings. Instead of using the method as a shortcut to developmental data, studies examining individual differences in accuracy and distortion and the factors that moderate them may inform us of the various meanings of retrospective data. PMID- 3042947 TI - [Promoting effect of the chymotrypsin inhibitor FK-448 on the intestinal absorption of insulin in rats]. PMID- 3042948 TI - [Acquired immunodeficiency syndrome (AIDS) and human immunodeficiency virus (HIV), and chemotherapeutic approaches to AIDS]. PMID- 3042949 TI - [Current status and possible future of antitumor drugs. With special reference to pharmacology of Oriental medicine]. PMID- 3042951 TI - Naviculocuneiform coalition. PMID- 3042950 TI - [Regulation of synthesis of proteins and nucleic acids by polyamines]. PMID- 3042953 TI - Pseudo axial length increase after silicone lens implantation as determined by ultrasonic scans. AB - Repeat A-scans of 45 patients who had silicone intraocular lens (IOL) implantation indicated an apparent increase in the axial length of the eyes. The average increase of 1.045 mm over the original axial length was noted when using the pseudophakic mode for the axial length determination. The actual velocity of sound in eyes with a silicone IOL is 1,486 M/sec compared to 1,548 M/sec in an eye with a polymethylmethacrylate implant. This finding is clinically significant when one is comparing the axial length of two eyes, one of which is pseudophakic. A 1 mm difference in axial length can cause serious doubts about the validity of one or the other of the scans if the type of implant is not known. The velocity of RMX-3 silicone was determined to be 1,049 M/sec. PMID- 3042952 TI - Imipenem/cilastatin: its use in the treatment of foot infections in the compromised host. PMID- 3042955 TI - Modified temporary keratoprosthesis in the triple procedure: a new surgical technique. AB - We present a new surgical technique for use in the triple procedure. After trephining the cornea, we sutured a temporary keratoprosthesis made of silicone rubber into the corneal bed. The prosthesis provided a clear view with excellent depth of focus. By creating an artificial anterior chamber, the operative procedure resembled a normal extracapsular cataract extraction procedure. The keratoprosthesis made complete aspiration of all cortical material easier. PMID- 3042954 TI - Age-related macular degeneration and its management. AB - Age-related macular degeneration is a major cause of vision loss in older individuals. The clinical picture and pathogenesis of age-related macular degeneration is reviewed. Present treatment modalities for subretinal vascularization in exudative-type macular degeneration and their limitations are discussed. The role of low-vision aids in providing magnification for reading vision is described. The role of intraocular lens implantation is discussed, as well as the newly developed intraocular lens which, with the addition of a plus lens, functions as a Galilean telescope to provide magnification for near vision. PMID- 3042956 TI - Closed vitrectomy technique during penetrating keratoplasty. PMID- 3042957 TI - Fibronectin and insulin for corneal surface defect phenomenon. PMID- 3042958 TI - Enhancement of the therapeutic effect of cephalosporins in experimental endocarditis by altering their pharmacokinetics with diclofenac. AB - We studied the effect of a nonsteroidal anti-inflammatory drug, diclofenac, in rabbits on the kinetics of three cephalosporins: cefotiam, cefmenoxime and ceftriaxone, and compared the antibacterial effect of these antibiotics, given alone or with diclofenac, in experimental endocarditis. Diclofenac significantly increased (P less than .05) the area under the curve in tissue-cage fluid of ceftriaxone and cefotiam-treated animals, and the terminal half-life of ceftriaxone in their sera (3.45 +/- 0.4 vs. 2.8 +/- 0.5 hr). Diclofenac reduced urinary excretion of cefotiam only. Cefmenoxime pharmacokinetics remained unchanged by diclofenac. The alteration of ceftriaxone kinetics appeared to be due to nonrenal mechanisms and could suggest reduction of biliary excretion. In Escherichia coli endocarditis, diclofenac enhanced the concentration (P less than .05) of cefotiam (23 +/- 16 vs. 8.9 +/- 5 micrograms/g) and ceftriaxone (13.2 +/- 3 vs. 8.5 +/- 4 micrograms/g) in infected vegetations, but not that of cefmenoxime. The antibacterial effect of ceftriaxone increased with diclofenac (5.5 +/- 1 vs. 7.2 +/- 1 log10 colony forming unit/g of vegetation). In vitro, neither protein binding to rabbit serum proteins nor intrinsic activity on the E. coli strain of each antibiotic was modified by diclofenac. These results suggest that anti-inflammatory drugs could increase antibiotic efficacy by altering their pharmacokinetics. The renal and nonrenal site of interaction may be involved for drugs belonging to the same class. Results obtained in tissue-cage fluid were predictive of the interference at the infected site. PMID- 3042959 TI - Centric relation and the treatment position in rehabilitating occlusions: a physiologic approach. Part I: Developing an optimum mandibular posture. PMID- 3042961 TI - Evaluation of shear strength at the cement-endodontic post interface. AB - The strength characteristics of the cement-implant interface were evaluated for smooth-tapered, threaded, and porous-surfaced endodontic implants with the use of different cements. Specifically, tensile and torsional shear strengths were measured for zinc phosphate, polycarboxylate, glass-ionomer, silicophosphate, and AH-26 cements. The results indicated superior shear strength characteristics for threaded endodontic implants on axial loading. However, this strength was diminished when torsional forces were applied. Porous-surfaced endodontic implants showed strong resistance to both axial and torsional loading. PMID- 3042962 TI - All-porcelain anterior fixed partial denture: a preliminary report. PMID- 3042960 TI - Long-term monitoring of microleakage of dental cements by radiochemical diffusion. AB - Radioactive 14C sucrose was found to be an ideal marker for microleakage because it did not penetrate tooth tissue, dental cement, or mounting resin. The main finding is that the adhesive cements--the glass-ionomer and polycarboxylate--are significantly more effective at preventing microleakage than are the traditional phosphate cements--silicate and zinc phosphate. The differences can be as high as two orders of magnitude. The adhesive cements provide almost perfect and reliable seals. By contrast, the nonadhesive cements are erratic sealants with most of the restorations leaking. PMID- 3042963 TI - Porous alumina for free-standing implants. Part I: Implant design and in vivo animal studies. PMID- 3042964 TI - Report of the Committee on Scientific Investigation of the American Academy of Restorative Dentistry. PMID- 3042966 TI - Simplified technique for boxing irreversible hydrocolloid impressions. PMID- 3042965 TI - Soft surface cast for mounting processed dentures. PMID- 3042967 TI - Fixed partial denture design and construction for missing mandibular molars by using an osseointegrated implant for an abutment. AB - This article presented a technique in the design and construction of a fixed partial denture by using a Synthes type E, ITI osseointegrated implant as a distal abutment in the mandibular arch. The technique may be altered to accommodate other forms of implants and uses methods and materials familiar to most dentists. PMID- 3042969 TI - Overdentures with metal occlusion to maintain occlusal vertical dimension and prevent denture fracture. PMID- 3042968 TI - Distribution of occlusal forces along unilateral posterior two-unit cantilever segments in cross-arch fixed partial dentures. AB - Axially directed occlusal forces over unilateral posterior two-unit cantilever segments of cross-arch fixed partial dentures were measured during natural functioning by using built-in transducers, one in each cantilever unit. The mean local maximal occluding and maximal chewing forces were significantly larger over the first (124 N and 64 N) than over the second (21 N and 29 N) cantilever unit. The average intraindividual ratio between the forces over the first and second cantilever unit amounted to 12:1 for maximal occlusion and 3:1 for maximal chewing. Despite the smaller mean total maximal chewing (92 N) than mean total maximal occluding cantilever force (145 N), the resulting axially directed mean bending moments in the joint between the distal abutment crown and the cantilever segment did not differ significantly. This is explained by the larger mean maximal chewing (29 N) than mean maximal occluding (21 N) force over the second cantilever unit. This demonstrated that not only the magnitude of occluding and chewing forces over cantilever segments but also their distribution along the cantilevers is of importance for the magnitude of functional stress created in cantilever fixed prosthesis. PMID- 3042970 TI - Using a syringe to make void-free casts from elastomeric impressions. AB - A syringe can be used to place dental stone into the deepest part of an impression, filling it from the bottom up, expressing air as the impression fills. PMID- 3042971 TI - Internally weighted mandibular dentures. AB - This procedure permits acrylic resin to almost completely surround the metal that may be used to add weight to a mandibular denture. PMID- 3042972 TI - Effect of chilled monomer on working time and transverse strength of three autopolymerizing acrylic resins. AB - Samples of three commercially available poly(methyl-methacrylate) autopolymerizing resins were made by using room temperature and chilled monomer. Working and setting times and transverse strengths were determined. The following conclusions can be drawn: 1. Working and setting times increased approximately 2 to 4 minutes when chilled monomer was used. This could be advantageous in certain situations. 2. Transverse strength decreased approximately 17% with the use of chilled monomer. The clinical significance of this decrease is not known. PMID- 3042973 TI - A technique for preparation of lingual rest seats in light-cured composite. PMID- 3042974 TI - Bonded composite rests: a fabrication method. PMID- 3042975 TI - A bifurcated ceramometal crown. PMID- 3042976 TI - Tooth preparation designs for resin-bonded fixed partial dentures related to enamel thickness. AB - Some basic principles for preparation of teeth to receive resin-bonded retainers is presented. Proper preparation of tooth surfaces will enhance retention and reduce overcontouring of the completed prosthesis. PMID- 3042977 TI - Management of Class III furcally involved abutments for fixed prosthodontic restorations. AB - Class III furca-involved molars provide formidable challenges, but the literature suggests that such teeth can be successfully treated. Under specific circumstances described in this article, reasonable longevity may be expected when advanced furcal involvement of abutments has been treated with (1) open-flap root debridement, (2) root separation with metal furcal replacement, (3) root resection as a conventional FPD or splint abutment, or (4) root resection as a vertical stop for a fixed cantilever prosthesis. Well-controlled maintenance therapy is essential for success. PMID- 3042978 TI - An evaluation of four techniques for condensation of three opaque porcelains. AB - Opaque porcelain applied with the brush and hand vibration techniques resulted in smaller and fewer entrapped air bubbles within the fired opaque. PMID- 3042979 TI - Enamel reduction and the bond strength of resin-bonded retainers. AB - The authors investigated the shear bond strength of etched enamel retainers bonded to enamel reduced by different amounts. The depth of reduction in this study had no influence on bond strength. PMID- 3042980 TI - A comparative study of the retentive capacity of dental cementing agents. AB - Although in vitro tests do not always duplicate in vivo conditions, they can offer comparative values. An in vitro test was done to evaluate the tensile strength of three types of cementing agents for complete crowns. PMID- 3042981 TI - Shear strength of composite bonded to laser-pretreated dentin. AB - As research progresses, laser energy moves closer to acceptable usefulness. Laser application to prepare dentin creates a more retentive surface for composite bonding. PMID- 3042982 TI - Effect of variation in powder-to-liquid ratio of zinc phosphate cement on the retention of posts. AB - This investigation revealed that lowering the powder/liquid ratio of zinc phosphate cement reduced the retention of posts. PMID- 3042983 TI - Veterans Administration Cooperative Studies Project No. 147. Part V: Interrater results of precementation evaluations of metal-ceramic crowns cast from alternative alloys. PMID- 3042984 TI - Reproducibility of the vertical dimension of occlusion with an improved measuring gauge. AB - An improved gauge using an eyeglass frame, the TOM gauge, was devised. The reproducibility of the record of vertical dimension with this gauge was evaluated through repeated measurements on subjects having a definite centric stop with the natural dentition. Because of the stabilization provided by the frame and the reference point on the apex nasi, the TOM gauge showed excellent reproducibility of the record compared with the conventional gauges. The TOM gauge can be expected to significantly reduce the risk of errors in measuring the vertical dimension of occlusion especially in complete denture fabrication. PMID- 3042985 TI - Model system for the in vitro testing of a synthetic histidine peptide against Candida species grown directly on the denture surface of patients with denture stomatitis. AB - The denture surface provides a nidus for the growth of microbial species that act to initiate, aggravate, and maintain clinical disease. The present investigation describes the development of a model system for the testing of the effectiveness of agents against these microbial species inhabiting the denture surface. It was observed through in vitro growth patterns that the model permitted the testing of representative samples of the microbial flora. Poly-L-histidine was observed to inhibit both Candida albicans and C. glabrata from growing from the denture surface into nutrient broth. Scanning electron microscopy of control and treated denture disks revealed that poly-L-histidine had either eliminated most microbial flora from the denture surface or had effected a noticeable distortion of those Candida blastospores still present on the surface. From microbiologic studies, it appeared that poly-L-histidine had inflicted direct but not lethal damage to the still-attached distorted blastospores because the latter were still able to promote growth in agent-free broth. The antifungal effects of poly-L-histidine were observed to be dependent on the concentration of the polypeptide. The data obtained were consistent for all of the patients regardless of their denture stomatitis classification. PMID- 3042986 TI - A multicenter report on osseointegrated oral implants. PMID- 3042987 TI - Implant prostheses for convertibility, stress control, esthetics, and hygiene. AB - A method of connecting "fixed partial denture" prostheses to osseointegrated implant fixtures has been described. The advantages of this system of restoration for partially and fully edentulous mouths are that it is more effective in addressing the problems of (1) stress-control on abutments, (2) a back-up system for abutment failures, (3) esthetics, and (4) control of bacterial plaques around abutments. To accomplish this procedure, the application of convertible periodontal prosthesis techniques with modifications to some existing implant systems is undertaken. The disadvantages of this method seem insignificant when one considers the complexities and risks involved with the present array of implant prosthesis alternatives. Some patients and dentists might consider the necessity of the prosthesis being detachable as one disadvantage. In reality, the prosthesis can be used as a fixed restoration until the patient has fully adapted to the new proprioception and appearance. A large percentage of patients feel uncomfortable with the word "removable" because it immediately creates a perception of unsightly metallic clasp display, palatal coverage, tongue interference, and negative body image. The use of the term "detachable" coupled with the doctor's offer to perform this task for the patient "whenever necessary" will usually relieve the patient's anxiety and allow the treatment to proceed. Once neuromuscular and esthetic adaptation have occurred and the patient has accepted the prosthesis, daily detaching and home-care hygiene by the patient will follow without incident. Esthetic improvement is obvious (Fig. 3). PMID- 3042988 TI - Two unique clinical applications using acid-etched restorations. AB - Two "Maryland bridges" replacing anterior teeth fused to the facial surface of the abutments are described. PMID- 3042989 TI - An evaluation of temporomandibular joint radiographs. PMID- 3042990 TI - An approach to the neurology of aggression. AB - Aggressive behavior is controlled at multiple anatomical levels within the human brain. To illustrate hierarchical neural controls over aggression, we compare and contrast the roles of the hypothalamus, amygdaloid complex, and orbital prefrontal cortex in terms of their distinctive sensory inputs, effector channels, and principles of integration as deduced from observations in animals and man. We illustrate characteristic syndromes of human aggression resulting from hypothalamic, temporolimbic, or frontal cortical lesions. The application of this perspective to research on criminal violence is discussed. PMID- 3042991 TI - Blood pressure reactivity in children. AB - This paper reviews the relevant pediatric literature about blood pressure (BP) hyperreactivity to stress as a possible precursor of hypertension and coronary heart disease. Two prospective studies of children indicate that BP hyperreactivity predicts later development of hypertension. Several studies have identified correlates of BP reactivity. Race, obesity, Type A and family history of hypertension appear to be associated with BP reactivity in children. There appear to be both genetic and environmental influences, but relatively few psychosocial variables have been studied. There are significant methodological concerns in terms of defining the characteristics of the stressors and the stability and generalizability of responses to laboratory stressors. BP reactivity in children is a potentially important area of inquiry that has been understudied. PMID- 3042992 TI - Effectiveness of defenses: a significant predictor of cortisol excretion under stress. AB - Although stress theories assert that psychological characteristics influence illness through their effects on physiological reactions to psychosocial stimuli, it has been difficult to demonstrate substantial associations to support this contention. Effectiveness of defense (ED) is a clinical assessment based on emotional reaction to stress, disruption of physiological and social functioning, and the ability to mobilize additional defenses to deal with acute, superimposed stress. By convention, high scores on these assessments define ineffective defenses. In 6 of 7 samples studied to date, a significant ED-cortisol correlation was obtained (combined significance, p less than 0.00006). The ED cortisol correlation averaged r = 0.41 for the seven samples and evidence from two studies suggests that ED is particularly important during high stress. These findings indicate that ineffective defenses are associated with higher cortisol excretion and establish ED as a topic worthy of study in connection with psychosocial stress. Future research issues include determining the importance of individual components of the overall ED rating and ascertaining whether ED has other physiological correlates in addition to cortisol. PMID- 3042993 TI - Positive perimenstrual changes: toward a new perspective on the menstrual cycle. AB - Although many women report negative symptoms, the perimenstrual phase also is associated with enhanced mood and performance among some women. However, research on perimenstrual concomitants reflects a sterotypic negative bias that does not encompass the complexity of the phenomena. This paper tries to redress that balance by documenting the prevalence of positive perimenstrual changes. Overall, about 5-15% of women experience increased excitement, energy, and well-being in the perimenstrual phase. Many women also report increased activity, heightened sexuality, and improved performance on certain types of tasks during the perimenstrual phase. Future research should examine why some women report positive perimenstrual changes, the extent to which individual variations in hormone levels can account for differences in women's perimenstrual experience, and how much women differ in their responsiveness to changing hormone levels. The influence of menstrual-related beliefs and expectations on the changes a woman reports also needs to be clarified. PMID- 3042994 TI - Symptoms of insulin-induced hypoglycemia in normal subjects. AB - In order to better characterize the psychobiology of hypoglycemic symptom production, six normal subjects had physiological/biochemical and symptom ratings at 20, 30 and 40 min after six different doses of intravenous regular insulin (0.0, 0.5, 1.0, 2.0, 3.0 and 4.0 units, in a Latin-square design); subjects also indicated after each dose whether they believed they had received an active or an inactive substance. Choice response switched from inactive to active substance when plasma glucose fell to the mid to high 50s mg/dl (i.e. whole blood glucose of approximately 50), and plasma epinephrine levels rose to between 100 and 200 pg/ml. Adrenergic symptoms and 'weakness' were most strongly associated with choice; other symptom variables had weaker associations. Symptoms were more strongly correlated with epinephrine than glucose levels, but anxiety was not strongly correlated with the epinephrine increases. PMID- 3042995 TI - Somatic presentations of psychiatric illness in primary care setting. AB - We have tried to describe somatisation, not as a disease, but as a common and important human mechanism involving both doctor and patient. It is the single most common reason why psychiatric illness goes undetected in general medical settings, and it often occurs in conjunction with physical disease processes. The association with dysphoric affect has been recognised at least since George Cheyne 250 years ago, and the reason for this is that both anxiety and depression serve to amplify pains. However, it seems likely that somatisation can occur in the absence of dysphoria. Once it has been established, it is easy to see how it continues: it secures advantages from spouse, family and employers; and it tends to be encouraged by doctors--who differentially reward somatic symptoms. But why does it occur in the first place? We have argued that it seems to have three functions: First, it allows people who are unsympathetic to psychological illness, or who live in cultures where mental illness is stigmatised, to nonetheless occupy the sick-role while psychologically unwell. Second, it is blame-avoiding: instead of being responsible for the mayhem, one is cast in the role of the suffering victim. Finally, by reducing blame, it appears to save patients from being as depressed as they might otherwise have been. PMID- 3042996 TI - [Castleman's disease. X-ray computed tomographic aspect. Apropos of 2 mediastinal forms and a multicentric form]. AB - A multicentric form of Castleman's disease is described, this being a rare affection for which CT image characteristics have not, to our knowledge, been reported. Two cases of localized and one case of multicentric Castleman's disease are reported, and differential characters of these two forms with common histology: angio-follicular lymphoid hyperplasia, outlined. The localized form is found mainly in the mediastinum in young patients and follows a favorable course with recovery after exeresis. The diffuse form develops in later life and presents with severe systemic signs and a marked biological inflammatory syndrome Glands and viscera are affected and its course is grave, with mostly fatal relapses. Angiography suggests diagnosis when images show hypervascular lesions in lymph glands. CT scan imaging with contrast is very suggestive when glandular lesions are present that take up contrast strongly. This appearance should always raise the possibility of Castleman's disease. PMID- 3042997 TI - [Pulmonary mesenchymatous chondrosarcoma in children. Report of 2 cases and review of the literature]. AB - Pulmonary mesenchymatous chondrosarcoma was revealed in two 3-year-old boys by hemi-thorax opacities on standard images and a heterogeneous mass on CT scanning. A favorable course was obtained after chemotherapy and total excision. These are the first two cases of pulmonary localizations of mesenchymatous chondrosarcoma reported. PMID- 3042998 TI - [Calcified masses of the kidney. Apropos of 58 cases]. AB - Case reports were analyzed of patients with calcified renal masses observed in the department since 1968. Of the 65 radiologic reports reviewed, 7 were rejected since the course since diagnosis was unknown. Of the 58 case reports studied, 34 were of masses of certain diagnosis, 12 undetermined, 7 of masses in polycystic kidneys, 3 in tuberculous kidneys and 3 probably calcified hematomas. Analysis involved only those masses of proven diagnosis. Results confirmed the absence of specificity in favor of the cyst of peripheral character of calcifications: 33% of these masses were cancers. The existence of tissue calcification is synonymous of a solid mass, nearly always malignant (92% of cases). For peripherally calcified masses, arteriography was not sufficient to affirm benign nature of lesions, most of these masses having a particularly poorly vascularized or even avascular appearance. In these cases angiotensin was of special interest. Ultrasound imaging proved to be a reliable and perfectly sensitive examination. The presence of calcifications rarely interfered with study of tumoral contents. CT scan imaging and puncture biopsy were also perfectly sensitive and reliable examinations. Because of the high frequency of cancers in masses with peripheral calcification, all these masses should be surgically explored or at least punctured. Although a "benign" CT scan image appears sufficient to affirm the benign nature, this still requires more ample confirmation. PMID- 3042999 TI - [Tuberculous calcifications of the bladder wall in children]. AB - A case of tuberculous urinary bladder calcifications in a 13 year old boy is reported. The lesion is rare in adults and no case is published in the literature as occurring in children. PMID- 3043000 TI - Carcinoma of the vulva then and now. The 1987 ISSVD presidential address. PMID- 3043001 TI - Ultrasound does not diagnose ureteric obstruction. PMID- 3043002 TI - Testing the retrovirus hypothesis of manic depression and schizophrenia with molecular genetic techniques. AB - Crow's viral hypothesis of schizophrenia proposes that psychosis may be the result of mutagenesis caused by viral integration or transposition in human genomic DNA. Molecular genetic techniques can be used to systematically investigate this hypothesis. In a study of genomic lymphocyte DNA unexpected DNA polymorphisms which were probably insertions and deletions were found in specific human genomic retroviral (proviral) related sequences. However these changes were found exclusively in normal Icelandic individuals and are probably of evolutionary origin. The extent to which human retroviral insertion and deletion has taken place and the mobility of such sequences will help in understanding their evolutionary origin and might provide a source of polymorphic marker sequences that could be used in genetic linkage studies of disease. PMID- 3043003 TI - Anaesthesia and Wolff-Parkinson-White syndrome during infancy: a review. PMID- 3043005 TI - Total number of neurons and glial cells in human brain nuclei estimated by the disector and the fractionator. AB - Unbiased estimates of the total number of neurons and glial cells from central regions of grey matter in human brains are obtained using the disector principle in modifications which are unaffected by the histological processing of paraffin embedded tissue. Section thickness does not enter into the estimator and need not be known. An analysis of the contributions from all sampling levels to the variance of the estimator of total cell number has been used to design an optimimal sampling scheme. Using this method, a precise and unbiased estimate of the total number of neurons in a defined brain region can be obtained in less than 1 h. PMID- 3043004 TI - Benjamin Collins Brodie 1783-1862. PMID- 3043006 TI - Salmonella meningitis in infancy. AB - The mortality and morbidity of salmonella meningitis in infancy is high. For infants and especially those younger than 6 months of age with salmonellosis, close observation is needed because of increased incidence of salmonella meningitis. A minimum duration of five weeks of susceptible antibiotic therapy may be indicated to prevent relapse or reinfection of salmonella meningitis. The role of a carrier in relapse needs to be evaluated. PMID- 3043007 TI - Antimicrobial activity of isoquinoline alkaloids and their N-oxide derivatives. PMID- 3043008 TI - Unusual complications of Salmonella enteriditis group D infection. AB - A 25-year-old Syrian presented complaining mainly of fever, night sweats and nausea. He had 3 days earlier mild abdominal cramps and short-lived diarrhoea. On admission, he developed signs of deep vein thrombosis and blood and stool cultures showed Salmonella enteriditis infection. The patient was started on chloramphenicol and later showed acute abdominal signs. Laparotomy revealed intestinal perforation on the lower ileum. The case together with the experience in this hospital and elsewhere of Salmonella enteriditis infections are discussed, showing that two complications shown in this case are common for Salmonella typhi and paratyphi infections but are unusual for other Salmonella infections. PMID- 3043009 TI - Response of Kampuchean strains of Plasmodium falciparum to antimalarials: in-vivo assessment of quinine and quinine plus tetracycline; multiple drug resistance in vitro. AB - Forty-three patients were enlisted in the in-vivo test for sensitivity of Kampuchean strains of Plasmodium falciparum to quinine. The mean parasite density count on day 0 was 32,398 asexual parasites per microliter of blood. With a dosage of 1.5 g quinine base daily for 10 days the average parasite clearance time was 5.6 days, and the average duration of fever 3.4 days. The in-vivo test was evaluated at 7 and 10 days after the start of therapy. After 7 days only 16 patients were parasite negative by microscopic examination (S); 20 patients had an RI recrudescence, four patients responded at the RII level and three at the RIII level. Evaluating the in-vivo test at 10 days, the number of patients parasite negative increased to 18, the number of those with an RI level of resistance increased to 21, two patients gave an RII response and two had an RIII level of resistance. Twenty-two adult males were included in an in-vivo test of the sensitivity of P. falciparum to quinine plus tetracycline. The course of treatment was: quinine 3 x 500 mg daily for 10 days, tetracycline 3 x 500 mg for 7 days. Parasite density counts on day 0 averaged 11,393 asexual parasites per microliter of blood. The average parasite clearance time was 5.9 days, and the average duration of fever was 3.8 days. After 7 days of treatment, 81.8% of patients were parasite negative, while one patient had a recrudescence after 17 days (RI). Three infections were resistant at the RII level. By prolongation of the observations to day 10, the parasitaemia was cleared in all patients, i.e. all infections were sensitive to quinine plus tetracycline. Thirty-four patients with falciparum malaria were screened for in-vitro resistance to chloroquine, mefloquine and quinine using the WHO standard micro-test.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3043011 TI - Throat carriage of Streptococcus pneumoniae and Streptococcus pyogenes among infants and children in Zambia. PMID- 3043010 TI - Comparison of haemagglutination test, enzyme-linked immunosorbent assay and indirect immunofluorescence antibody test for determination of rubella immune status. AB - Comparative evaluations of immune status for rubella are described for the enzyme linked immunosorbent assay (ELISA), indirect immunofluorescence (IFA) and two standard haemagglutination inhibition tests (HAI kaolin; HAI heparin-MnCl2). In general, a reasonably good correlation was obtained between the level of rubella antibodies measured by the HAI (kaolin) test and by ELISA, but an appreciable proportion (15%) of ELISA positive specimens were encountered among HAI (kaolin) negative sera. All of these HAI negative, ELISA positive sera, except one were found to be positive for rubella antibodies by IFA. Neutralization test performed on this serum positive by ELISA only, confirmed the presence of protective rubella antibodies. Of all the tests evaluated ELISA appeared unequivocally to be the most sensitive test followed closely by IFA. The standard HAI (heparin-MnCl2) was more suitable than the HAI (kaolin), for the determination of immune status. Further, no linear relationship between single rubella virus HAI and ELISA values was observed. PMID- 3043012 TI - Vitamin E and human nutrition. PMID- 3043013 TI - The biomechanics of javelin throwing: a review. AB - In this paper, the scientific literature and that on the sports sciences relevant to javelin throwing is critically reviewed. This is particularly timely because of the change in the specification of the javelin for the men's event, which was introduced by the IAAF in 1986. A full discussion of the aerodynamics of the javelin is presented with due consideration of the change in pitching moment characteristics that the rules change had brought about. The uses and limitations of current computer programs for simulating javelin flight, in order to estimate optimal release parameters, are profiled. Consideration is also given to the effects of wind velocity, air density, javelin weight and the flutter and spin of the javelin on its flight. The review further considers the optimization of release parameters, drawing upon computer simulations and field-based data. The effects of release speed, release height, release angle, release angle of attack and release pitch rate are assessed. The javelin throwing technique is discussed in relation to cinematographically derived data, including an evaluation of experimental procedures. The importance to successful performance of the grip, the run-up and transition phases, the cross over and delivery strides are each reviewed. Finally, some prognoses as to the direction of future research into this complex throwing skill are offered. PMID- 3043014 TI - Echocardiographic dimensions in trained and untrained 12-year-old boys and girls. AB - The purpose of this study was to compare echocardiographically measured left ventricular (LV) dimensions of 85 trained 11-12-year-old athletes with 106 untrained children matched for skeletal age and fat-free mass. Training status for each group applied to the 3 years prior to the measurements. It was found that 12 min and 100 m runs demonstrated the superior athletic ability of the trained children, but there were no significant differences in LV internal diameters at diastole and systole, in LV posterior wall thickness, or in LV end diastolic volume and LV mass. These data indicate that little difference occurs in LV size between moderately trained and untrained 11-12-year olds or between boys and girls matched for fat-free mass and skeletal age. It is also evident that consistent but moderate training during late pre-adolescence has little effect on LV development. PMID- 3043015 TI - Sonographic estimation of amniotic fluid volume. Subjective assessment versus pocket measurements. AB - Amniotic fluid volume (AFV) estimation is an important part of routine obstetric sonography. Despite the clinical importance placed upon excessive or diminished AFV in pregnancy, there is little uniformity in the way it is estimated sonographically. We compared AFV estimations obtained using two commonly employed sonographic methods "subjective" visual assessment and amniotic fluid pocket measurements. Estimates obtained using both methods correlated closely. In addition, there was excellent intraobserver and interobserver agreement among estimates obtained using subjective criteria. This supports the belief that experienced observers tend to agree on the sonographic appearance of normal, excessive or decreased AFV. Methods for AFV estimation and potential pitfalls are discussed. PMID- 3043016 TI - Posterior urethral obstruction. Prenatal sonographic findings and clinical outcome in fourteen cases. AB - Fourteen cases of fetal urethral obstruction were reviewed retrospectively. The purpose of this study was to emphasize the following: 1) prenatal sonographic findings: 2) clinical outcome: and 3) associated congenital anomalies. Decreased amniotic fluid volume complicated 12 pregnancies (86%). A dilated posterior urethra was identified in nine fetuses (64%) and an enlarged bladder in 13 (93%). Evaluation of the fetal kidneys revealed hydronephrosis in 81%, increased parenchymal echogenicity in 73% and macroscopic renal cysts in 15%. There were seven live births, but only two neonates survived beyond 5 weeks. Pulmonary hypoplasia contributed to the five postnatal deaths. Associated congenital anomalies were noted at autopsy in six cases. PMID- 3043017 TI - Transvaginal ultrasound directed oocyte collection for in vitro fertilization: successes and complications. AB - From December 1985 to April 1986, 458 patients underwent 600 transvaginal ultrasound-directed oocyte pickups (TVOPU) in preparation for in vitro fertilization and embryo transfer at The Royal North Shore Hospital of Sydney. Collections (552) were performed under light general anesthetic and, more recently, 48 were performed with light sedation. At least one oocyte was collected in 98.7% of all TVOPUs. A total of 3117 follicles (greater than 12 mm) were aspirated and 2576 oocytes were collected. Ninety-two clinical pregnancies were established after embryo transfer in 474 cycles. Major operative morbidity was low, with three pelvic abscesses and three pelvic hematomas reported. The procedure was well accepted by patients, with only one expressing a preference for the laparoscopic method. TVOPU is now the preferred method of oocyte pickup in our unit. PMID- 3043018 TI - Vaginal sonography in ectopic pregnancy. A prospective evaluation. AB - Among 404 first-trimester pregnancies examined with vaginal sonography in a prospective study, there were 21 ectopic gestations. Considering only the initial scans, the endometrial canal showed a linear echo surrounded by an echogenic zone in 18 cases, but in three cases the uterine cavity demonstrated a small echo-free area representing blood. Free fluid within the cul-de-sac was seen in 17 patients. An adnexal tumor representing the extrauterine gestation, was detected in 19 cases. Fifteen of these masses exhibited a thick-walled ring characteristic of a gestational sac with a viable embryo in five cases and a yolk sac in one. Other cystic adnexal masses, such as corpus luteum cysts, seen in 14 of the 21 patients were not confused with the ectopic pregnancy. A correct tentative diagnosis of ectopic gestation was made in 18 patients (86%) after the initial scan and in 20 cases (95%) including four controls. There was one false-positive suspicion of ectopic gestation in a patient who actually had a spontaneous abortion. Interpretation of the sonographic image should generally be done in correlation with laboratory and clinical data. The results of the study indicate that vaginal sonography is a valuable diagnostic procedure in the evaluation for ectopic pregnancy. PMID- 3043019 TI - Sonography of the low transverse incision, cesarean section: a prospective study. AB - Using sonography, the uterine incision site was prospectively studied in 36 asymptomatic patients, two days after cesarean section. The findings were compared with those seen in 21 symptomatic, postcesarean patients. In the asymptomatic patients, the incision site was visualized as an oval symmetric region of distinct echogenicity interposed between the lower uterine segment and the posterior wall of the urinary bladder. In eight of the 36 asymptomatic patients, a small (less than 1.5 cm) round hypoechoic mass was present in or adjacent to the uterine incision and distinct from the normal incision. These probably represented insignificant hematomas. Of the 21 symptomatic patients, 17 had either a subfascial hematoma, a bladder-flap hematoma, or endometritis. Two were sonographically normal, and one showed a hematoma in the paracolic gutter. In the remaining patient, there was a 5-mm asymmetrically placed hypoechoic mass representing an insignificant hematoma. Significant bladder-flap hematomas were characteristically round, greater than 2 cm masses asymmetrically placed in or adjacent to the uterine incision. Using sonography, the normal appearance of the lower uterine incision can be distinguished from significant hematomas. PMID- 3043020 TI - Decreasing size of a congenital cystic adenomatoid malformation in utero. PMID- 3043021 TI - Mirizzi syndrome with common septum: ultrasound and computed tomography findings. PMID- 3043022 TI - Acardiac twin. Report of Doppler sonography. PMID- 3043023 TI - Growth factor production by Creutzfeldt-Jakob disease cell lines. AB - Creutzfeldt-Jakob disease (CJD), a progressive dementia of humans, is caused by an infectious agent that is closely related to the scrapie agent of sheep. Although the molecular nature of these "unconventional" agents is still a matter of speculation and controversy, even less is known concerning the mechanism(s) of their effects on the central nervous system. To gain insight into the cellular effects of these agents, we have examined a series of cell lines derived directly from CJD-infected hamster brain or produced from nontransformed rodent lines by exposure to CJD infectious fractions in vitro. These cell lines appear transformed by a variety of criteria and secrete growth factors into the culture medium. All CJD lines produce a factor that is like alpha-transforming growth factor (alpha-TGF). Conditioned medium from these CJD lines also stimulates the synthesis of glial fibrillary acidic protein in normal astrocytic cells in vitro. This effect is mimicked by purified alpha-TGF and platelet-derived growth factors. Further study of CJD-induced growth factor production may elucidate fundamental properties of these unconventional agents. PMID- 3043025 TI - Leads from the MMWR. Shigella dysenteriae type 1 in tourists to Cancun, Mexico. PMID- 3043024 TI - Multiple early transcripts and splicing of the Autographa californica nuclear polyhedrosis virus IE-1 gene. AB - The immediate-early IE-1 gene of Autographa californica nuclear polyhedrosis virus was cloned, and its nucleotide sequence was determined. Sequence analysis indicated that this gene would encode a protein of 582 amino acids with a predicted molecular weight of 66,822. Analysis of IE-1 gene expression during baculovirus infection identified two transcripts. One, 1.9 kilobases (kb), was expressed at constant steady-state levels throughout infection, whereas the other, 2.1 kb, was expressed only early in infection. Analysis of IE-1 cDNA clones demonstrated that the 2.1-kb transcript contained the entire 1.9-kb transcript (exon 1) plus an additional 5' end (exon 0). Genomic Southern analysis placed the exon 0 sequences on the EcoRI B fragment, 4 kilobase pairs upstream of exon 1. Sequencing of the upstream region identified an open reading frame whose 5' end was identical to the exon 0 sequences in the cDNAs. Examination of the genomic DNA sequences around the exon-exon junction revealed sequences similar to published consensus splice acceptor and donor sequences. This is the first example of splicing of any viral transcript during baculovirus infection. PMID- 3043026 TI - A piece of my mind. Commander. PMID- 3043027 TI - Allied health education and accreditation. PMID- 3043028 TI - Medical schools in the United States. PMID- 3043029 TI - Medical schools in Canada. PMID- 3043030 TI - The future of family practice. Implications of the changing environment of medicine. Council on Long Range Planning and Development in cooperation with the American Academy of Family Physicians. AB - The Council on Long Range Planning and Development of the American Medical Association has identified trends in the environment of medicine and the implications of these trends for specific medical specialties. This report considers the evolution of family practice as a specialty and its role in the future of health care delivery. As a specialty established less than 20 years ago, family practice has successfully surmounted several obstacles to achieve recognition within the medical community and among the public. However, the Council has identified new challenges and opportunities facing this specialty. In particular, the areas of graduate medical education, reimbursement, professional liability, and several health-related societal and ethical issues will pose challenges for and place constraints on family physicians. Family practice will encounter a number of opportunities in the evolving environment of medicine, due in part to demographic trends in the population and the growth in managed care. The Council concludes that, despite the challenges, the increasing demand for the services of family physicians has positive implications for the future of this specialty. PMID- 3043031 TI - Articular oxalate crystals and the taxonomy of gout. PMID- 3043032 TI - [Efficacy and safety of cefbuperazone in severe infections complicating hematologic diseases Hanshin Infection Study Group]. AB - Cefbuperazone (CBPZ) was administered to patients with severe infections complicating hematologic diseases to assess its efficacy and safety under such clinical conditions. Primary diseases in this series of 78 cases included; acute leukemia in 41 cases, chronic leukemia in 6 cases, other leukemia in 9 cases, malignant lymphoma in 13 cases, multiple myeloma in 3 cases, aplastic anemia in 5 cases and 1 other case. Types of infection included sepsis; proven or suspected, in 59 cases, pulmonary infection in 8 cases, upper respiratory infection in 5 cases, and other cases. CBPZ was infused by an intravenous drip method at a dosage of 4-8 g daily. Patients' ages ranged from 14 to 85 years. Clinical response to the CBPZ regimen was excellent in 24 cases, good in 22 cases, fair in 2 cases, and poor in 30 cases. Thus the overall efficacy rate (percentage of cases showing an excellent or good response) was 59.0%. Efficacy rates for individual types of infection were: documented sepsis 16.7%, suspected sepsis 58.5%, lower respiratory infection 62.5%, and upper respiratory infection 100%. CBPZ also proved to be effective in 61.0% of cases with a neutrophil count of less than 500/mm3 prior to therapy. Side effects encountered were diarrhea in 1 case, gastric discomfort in 1 case and hepatic dysfunction in 5 cases. These side effects, however, were not dose-related, and none were serious. These results indicate that CBPZ has a high therapeutic efficacy even in patient with compromised immunodefenses. PMID- 3043034 TI - [Aztreonam]. PMID- 3043033 TI - [Clinical studies on effect of cefbuperazone on surgical infections]. AB - We conducted clinical studies on the efficacy and safety of cefbuperazone (CBPZ) on surgical infections with the following results. 1) In evaluable 32 patients, CBPZ was evaluated to be clinically effective in 23 (71.9%) and the efficacy rate was better in those who were administered 4 g/day of CBPZ (87.5%, 14/16) than in those who were given 2 g/day (57.1%, 8/14). 2) Antibacterial activity of CBPZ was evaluated in 41 isolated bacterial strains. Pathogen eradication rate by bacterial species was 75.6% (31/41). CBPZ exerted excellent antibacterial effects on Escherichia coli (100%, 8/8) and anaerobic bacteria such as Bacteroides (88.9%, 8/9). Resistant bacteria to CBPZ were Enterococcus faecalis and Pseudomonas aeruginosa. 3) No serious side effects were noted in any of the 34 patients who entered in this study. Abnormal laboratory test results were noted in 2 patients (5.9%) and they were transient elevation of transaminases and alkaline phosphatase. From the results shown above, CBPZ appears to be a highly useful antibiotic for the treatment of surgical infection. PMID- 3043035 TI - [Laboratory and clinical studies on cefotetan in respiratory tract infections]. AB - Seventy five patients with respiratory infections, including 40 cases of acute pneumonia, 33 cases of secondary infection after chronic pulmonary diseases and 2 cases of pulmonary abscess, were treated with cefotetan (CTT, Yamatetan) by drip infusion in order to evaluate its clinical efficacy. The overall rate of effectiveness was 83.8%. CTT was examined comparatively with other beta-lactam antibiotics for antibacterial activity on clinically isolated strains of 3 major respiratory pathogens including Haemophilus influenzae, Branhamella catarrhalis and Streptococcus pneumoniae. Minimum inhibitory concentrations (MIC's) of CTT on H. influenzae were less than 3.13 micrograms/ml regardless of the production of beta-lactamase by these organisms. As to B. catarrhalis, CTT also exerted an antibacterial activity enough to control the proliferation of all the strains at a level of 1.56 micrograms/ml. Against S. pneumoniae, on the other hand, CTT exhibited the lowest activity of all the drugs tested but still showed MIC's of 3.13 micrograms/ml or less. Drip infusion of CTT at a dose of 2 g brought about an average maximum blood concentration of 342 +/- 25.7 micrograms/ml and an average half-life in blood of 2.48 +/- 0.41 hours Maximum sputum concentration of the drug, however, was variable among the cases tested, ranging from 0.40 to 1.80 micrograms/ml. Side effects of the drug were observed in 5 cases or 6.7%. Four of them had some allergic symptoms; i.e., pyrexia and eruption. One patient was especially diagnosed as possible drug-induced interstitial pneumonia during the treatment with the drug. The diagnosis was confirmed by transbronchial lung biopsy and lymphocyte blastogenesis by CTT in vitro. As to abnormal laboratory findings, blood transaminases were elevated during drug administration in 13 cases or 17.3%, but were reduced back to the normal level after the withdrawal of the drug. PMID- 3043037 TI - [A histologically-verified triple cancer--report of a rare case involving a primary brain tumor]. AB - Histologically-verified triple cancers that include a malignant brain tumor are rare. According to the Japan Autopsy Annuals, only 8 cases since 1958 have been so far documented. A case combining a malignant melanoma, a medulloblastoma, and a thyroid cancer is herein presented, along with a review of the literature. In March, 1983, a 27-year-old female who, 7 years prior to admission, had had malignant melanoma on the right hand removed, complained of headaches and vertigo. A CT scan revealed a right cerebellar mass lesions, which subsequently proved to medulloblastoma. Three years later, a struma was found and subtotal thyroidectomy revealed a papillary adenocarcinoma. PMID- 3043036 TI - [A study on the bactericidal action of aspoxicillin against Escherichia coli]. AB - In an attempt to clarify the role of a side chain, N4-methyl-D-asparagine, of aspoxicillin (ASPC) in the antibacterial action, we examined the bactericidal activity of dehydroxyaspoxicillin (AB-ASPC) and its affinity for the penicillin binding proteins (PBPs) of Escherichia coli using piperacillin (PIPC), mezlocillin (MZPC) and apalcillin (APPC) as the reference penicillins. ASPC and AB-ASPC showed high bactericidal activities against E. coli K-12 even when a large inoculum size (2 x 10(8) CFU/ml) was used. The observation of these cultures with a phase contrast microscope revealed that E. coli cells lysed after the formation of spheroplast-like or bulged structures. On the other hand, PIPC, MZPC and APPC converted the cells to long filaments, but did not show lytic action in the range of the concentrations used. These morphological changes were also observed with a scanning electron microscope. Superior bacteriolytic activities of ASPC and AB-ASPC were further shown by measuring 'triggering' autolytic activity by the penicillins. The release of labeled murein from E. coli chi 1776 after the exposure to ASPC or AB-ASPC was clearly greater than those caused by the reference penicillins. ASPC showed affinity for PBPs of E. coli K 12, 1A, 1Bs, 2 and 3, and its affinity pattern resembled the one obtained with ampicillin (ABPC). AB-ASPC behaved in a fashion similar to ASPC, although its affinities for PBP 1A and 1Bs were lower and that for PBP 3 was slightly higher. These observations suggest that the highest bactericidal activity of ASPC against E. coli with lysis among the acyl-ureidopenicillins tested is due to N4-methyl-D asparagine in the side chain of ASPC. PMID- 3043038 TI - [Two cases of renal cancer detected by ultrasonic scan during mass screening and subsequent MRI]. AB - Reported herein are the cases of two woman, aged 47 and 69, respectively, who were each found to have a right renal carcinoma, both carcinomas discovered incidentally by an abdominal ultrasonic scan during a mass screening, with no other signs or symptoms of renal cancer being indicated. Following clinical diagnostic verification, using X-ray CT, and an MRI, radical nephrectomies were performed. In each instance, the pathohistological diagnosis was renal cell carcinoma, pT2N0M0V0. The clinical imagings, including the MRI, are presented, and the diagnostic techniques used in this imaging and the screening modality in cases of asymptomatic renal cancer are discussed. PMID- 3043039 TI - [Pulse methylprednisolone therapy in children with idiopathic thrombocytopenic purpura]. PMID- 3043040 TI - [Sequential examination of anti-human immunodeficiency virus (HIV) antibodies by Western blot analysis and immunity in hemophiliacs]. PMID- 3043041 TI - [A study of B-triple V (behenoyl-ara C, etoposide, vincristine, vinblastine) in acute leukemia]. PMID- 3043042 TI - [Acute mixed leukemia (AMixL) with both myeloid and lymphoid phenotypes: selection of treatment for AMixL]. PMID- 3043043 TI - [A case of malignant lymphoma occurring subsequent to Evans' syndrome]. PMID- 3043044 TI - [Endothelial injury as an initiating factor: role of endothelial cells and arterial endothelial cells in atherosclerosis]. PMID- 3043045 TI - [Pathogenesis of atherosclerosis and blood platelets: pathogenesis of atherosclerosis and etiologic role of platelet TXA2 synthesis]. PMID- 3043046 TI - [Regression and therapy of atherosclerosis: trends in the development of new therapeutic agents for arteriosclerosis in Japan and foreign countries]. PMID- 3043047 TI - [Bacterial flora of the large intestine]. PMID- 3043048 TI - [Structure of receptors and expression of anti-receptor antibodies. a. Insulin receptor antibodies]. PMID- 3043049 TI - [Clinical study of autoimmune diseases: recent trends--with special reference to progress in immunological tests and therapeutic methods. Organ-specific autoimmune diseases. 2) Type I and insulin resistance B type diabetes]. PMID- 3043050 TI - [Prostaglandin J2--anti-tumor activity and mechanism of action]. PMID- 3043051 TI - [Realities and therapy of radiation injuries resulting from the accident at the Chernobyl Nuclear Power Plant]. PMID- 3043052 TI - [Interruption of cell proliferation and the TGF-beta regulatory protein]. PMID- 3043053 TI - [Advance in apolipoprotein study]. PMID- 3043054 TI - [Apolipoprotein E and atherosclerosis--lipid metabolism in foam cell]. PMID- 3043055 TI - [Laboratory testing on cerebrospinal fluid-cytodiagnosis, tumor markers and alkaline phosphatase-binding immunoglobulin]. PMID- 3043056 TI - [Analysis of peripheral blood lymphocyte subsets--concerning a routine laboratory test]. PMID- 3043057 TI - [Determination of adrenal steroids by fluoroimmunoassays]. PMID- 3043058 TI - [Application of chemiluminescent analysis to the immunoassay detection system]. PMID- 3043059 TI - [Bacterial infection]. PMID- 3043060 TI - [Detection of treponema pallidum specific antibodies by high performance liquid chromatography and it's clinical evaluation]. PMID- 3043061 TI - [Quantitative bacteriological analysis of expectorated sputum by spiral system]. PMID- 3043062 TI - [Influence of media and proteinase (CAPP) on the growth of Candida albicans: mechanism of changes in pH in media]. PMID- 3043063 TI - [Cell kinetics in human anagen hair and hair follicles studied with bromodeoxyuridine (BrdU) and anti-BrdU monoclonal antibody]. PMID- 3043065 TI - [Antinuclear antibodies (ANA) in healthy subjects]. PMID- 3043066 TI - [Studies on immunoglobulins in pancreatic juice from patients with pancreatitis]. PMID- 3043064 TI - [Application of ultrasonic diagnostic equipment in dermatology--measurement of skin thickness]. PMID- 3043067 TI - Antinephritic effect of prostaglandin E1 on serum sickness nephritis in rats (5). Effect of PGE1 on disposal of heat-aggregated bovine serum albumin in the glomerulus. AB - In the present study, we investigated whether prostaglandin E1 (PGE1) could accelerate the disposal of heat-aggregated BSA (a-BSA) in the glomerulus by mesangial cells and/or resident mesangial cells. ICR mice were injected i.v. with 90 mg/100 g B.W. of a-BSA 3 times at 4-hr intervals. Kidneys were isolated at various times after the first injection of a-BSA. The location of a-BSA in the glomerulus was then detected by immunohistochemical staining and immunofluorescence. A-BSA was detected in the mesangium and along capillary walls by both techniques. The amount of glomerular a-BSA increased with time, attaining a peak about 12 to 14 hr after the first administration of a-BSA and then disappeared by 36 hr after. The mice injected with a-BSA 3 times received 150, 200, 300 and 400 micrograms/mouse of PGE1, s.c., and 200 and 400 micrograms/mouse of PGE2 or PGF2 alpha, s.c., at 12 hr; and their kidneys were isolated at 16 hr. The mice with 300 and 400 micrograms/mouse of PGE1 had 34.3% and 37.6% less a-BSA than the control mice, respectively. Additionally, the mice with 400 micrograms/mouse of PGE2 had 63.2% less a-BSA than the control mice. However, PGF2 alpha failed to reduce glomerular a-BSA to a level less than that of the control. In conclusion, we confirmed that PGE1 accelerates breakup of macromolecules by mesangial cells and/or resident mesangial cells in the glomerulus. PMID- 3043068 TI - Experience with 247 living related donor nephrectomy cases at a single institution in Japan. AB - There is currently much concern over the morbidity and mortality of donors undergoing nephrectomy for living related renal transplants. Between April, 1970 and July, 1986, 247 cases of living related renal transplants were performed at the Second Department of Surgery, Kyoto Prefectural University of Medicine. The average age of the donors was 50.3 +/- 9.7 years, 81 per cent of the donors being parents of the recipients. Minor abnormalities which did not affect the donors suitability were found in 71 cases. Nephrectomies were performed extraperitoneally in all cases. Peri-operative complications, including wound complications in 13 cases, urinary infection in 12 cases and pulmonary complications and arrhythmia in 4 cases, were considered to be minor in nature. A variety of renal function tests, carried out two weeks after nephrectomy revealed normal levels, although they had become slightly worse than those estimated pre operatively. Long-term sequelae in the follow-up period from 18 months to 16 years and 2 months, was studied on 124 donors who answered questionnaires. Currently, there are 5 late deaths, none of which are directly related to the nephrectomy. Of the 124 donors, 85.5 per cent stated that there had been no change in their physical states following surgery. Pain or a feeling of discomfort at the wound site was reported by 10 donors (8.1 per cent) and hypertension was observed only in 3 (2.4 per cent). No major complication directly related to the donor nephrectomy was found, except for one case of incisional hernia.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3043069 TI - Histologic and hemodynamic investigation on canine lung allografts. AB - Histologic and hemodynamic investigations were performed on 26 mongrel dogs that underwent left lung allotransplantation. All dogs received cyclosporin A and azathioprine as immunosuppressants and were divided into two groups. In the 10 Group I dogs, no preservation was performed and the animals were followed until death. The 16 Group II dogs were subdivided into two groups; Group II-A dogs received the transplantation after 6 hours preservation and Group II-B dogs received it after no preservation. All dogs in Group II were sacrificed within 14 days following the transplantation. Seven of the dogs in Group I survived for over 90 days. The major histologic findings of Group I were pneumonitis, fibrotic interstitial changes and pleural thickening. Atypical pneumocytes were observed in three dogs, however structural changes of the bronchioles, suggesting obliterative bronchiolitis, were obscure. In Group II, 75 per cent of the dogs demonstrated the histologic features of rejection and early rejection was seen in a few dogs. Under electron microscopy, vascular wall damage was indicated by swollen endothelial cytoplasm and disrupted basement membranes. Small lymphocytes accumulating around the vessels showed lymphoblastic figures with rich intracytoplasmic organellae. The mean pulmonary arterial pressure measured by right pulmonary artery (PA) occlusion was elevated just after the transplantation in Group II, but decreased on the 7th day. The mean PA was again elevated on the 14th day in those dogs in which diffuse mononuclear cell cuffing was demonstrated. We consider that the PA-occlusion test can be used for determining the degree of rejection. PMID- 3043072 TI - Segmental autotransplantation of the pancreas after total pancreatectomy for advanced periampullary carcinoma--a case report. AB - A case is reported here in which segmental autotransplantation of the pancreas was performed after total pancreatectomy for advanced periampullary carcinoma in an attempt to preserve pancreatic endocrine function. The postoperative course was uneventful. The requirement of insulin decreased after the operation and the daily profile examination, done 5 months following surgery, showed a permissive fluctuation of blood sugar levels without insulin injection. Thus, segmental autotransplantation of the pancreas offers a method of preserving pancreatic endocrine function after total pancreatectomy for periampullary carcinoma in selected patients. PMID- 3043070 TI - Congenital mesenteric arterio-portal fistula: report of a case. AB - A male patient with an arterio-portal fistula resulting from a mesenteric arteriovenous malformation, who developed portal hypertension and liver cirrhosis, is presented herein. The malformation was considered to be congenital in origin and its location made any ablative surgical procedure impossible. Such alternative treatments as ligation of the afferent arteries, followed by transarterial embolization were therefore given, but both were unsuccessful. We also present a review of the literatures of mesenteric arteriovenous fistula. Radical surgical approach for this rare entity is proposed. The case reported here as related to mesenteric arteriovenous communications of congenital origin is the seventh such case published, and the first which was ever found to be located in the trunk of the superior mesenteric artery. PMID- 3043071 TI - Pulmonary varices: a case report and review of the literature. AB - A case of a 47-year-old woman with pulmonary varix is reported herein. Saccular dilatation of the inferior pulmonary vein resembled a pulmonary perihilar mass which could not be palpated at the time of thoracotomy. Aneurysmal dilatation of the pulmonary vein, otherwise known as pulmonary varix, is rare. Only 71 such cases, including 17 cases in Japan, have been reported. Pulmonary varices may be classified into three types, namely: saccular type, tortuous type and confluent type. Most of the varices seen in patients with valvular disease have been of the confluent type (62 per cent), however tortuous type varices have also been seen in some cases (19 per cent). Pulmonary venous hypertension may be one of the major causes of confluent type pulmonary varices as regression of pulmonary varices after mitral valve replacement has been reported. None of the saccular type cases, however, were accompanied by valvular disease. This indicates that local factors may also be an important cause of saccular type varices. PMID- 3043073 TI - [Tricuspid annuloplasty with polytetrafluoroethylene sheet]. PMID- 3043074 TI - [Infections due to Mycobacterium simiae, Mycobacterium asiaticum and Mycobacterium shimoidei]. PMID- 3043075 TI - Insulin release from pancreatic islets: effects of CRF and excess PTH. AB - Insulin secretion may be impaired in chronic renal failure (CRF) and available data suggest that this abnormality may be related to the state of secondary hyperparathyroidism of renal failure. We directly measured insulin release from isolated islets of Langerhans obtained from normal rats, CRF-control and CRF-PTX (parathyroidectomized) rats, and parathyroid hormone (PTH)-treated animals. Both early and total glucose-induced insulin release from islets of CRF-control were markedly and significantly (P less than 0.01) lower than from islets of normal rats. Insulin release from islets of CRF-PTX rats was significantly (P less than 0.01) higher than that from islets of CRF-control rats, and not different from insulin release from islets of normal rats. Forskolin and IBMX, which cause a rise in cAMP, significantly stimulated glucose-induced insulin release from islets of normal, CRF-control and CRF-PTX rats, but the increments from baseline were not significantly different between the three groups. Both early and total insulin release from islets obtained from PTH-treated rats with normal renal function were markedly and significantly (P less than 0.01) lower than values obtained from normal rats. Calcium contents of the pancreas of CRF-control and PTH-treated rats were significantly (P less than 0.01) higher than that in pancreas of normal rats and CRF-PTX animals, and values in the latter two groups of animals were not significantly different. The results show that: 1) CRF impairs insulin release from pancreatic islets; 2) this abnormality is reversed by prior parathyroidectomy; and 3) hyperparathyroidism induced by PTH-treatment in normal rats impairs insulin release from pancreatic islets.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3043076 TI - The anatomy of the renin-secreting cell in adult polycystic kidney disease. AB - We used a human renin antiserum and an immunoperoxidase method to investigate the distribution of renin-containing cells in 19 adult polycystic kidneys: 9 autopsy and 10 nephrectomy cases. These cells were present in residual normal kidney, in scarred renal parenchyma and in area of fibrous tissue. They were situated mainly in juxtaglomerular apparatuses (JGAs) and in the walls of arteries and arterioles. A semi-quantitative analysis showed hyperplasia of JGA renin containing cells in the untreated autopsy cases. In both groups there was an abnormal distribution of renin-containing cells; only 50% were located in the JGAs the remainder were mainly in the walls of small arteries. These were often thin, attenuated vessels in the walls of cysts. Cells containing renin were also found isolated in fibrous tissue separate from the arterial tree. This abnormal location of the renin-containing cells suggests that they may respond to different stimuli than those in normal kidneys, and their abnormal distribution could affect both the intrarenal action of renin and also its release into the blood. PMID- 3043077 TI - Controlled plasma exchange trial in acute renal failure due to multiple myeloma. AB - We studied 29 patients affected by acute renal failure due to multiple myeloma with Bence-Jones proteinuria greater than 1 g/day to evaluate the effectiveness of plasma exchange in the treatment of severe myeloma nephropathy. Renal failure was severe enough to require dialysis in 24 cases, while the remaining 5 patients showed serum creatinine levels greater than 5 mg/dl. The patients were randomly allocated to Group I (15 patients undergoing plasma exchange together with corticosteroids, cytotoxic drug, hemodialysis only when needed) or to Group II (14 patients undergoing peritoneal dialysis together with corticosteroids and cytotoxic drug). In Group I Bence-Jones proteinuria decreased dramatically (P less than 0.01) with a significant increase in urine output (P less than 0.001), while Group II presented a slight reduction in Bence-Jones proteinuria without a significant increase in daily diuresis. Thirteen out the 15 Group I patients recovered renal function reaching serum creatinine levels less than or equal to 2.5 mg/dl in most cases. Only two patients in Group II improved renal failure well enough to stop dialysis. The one-year survival rate was significantly higher in Group I (66%) than in Group II (28%, P less than 0.01). We conclude that plasma exchange associated to chemotherapy rapidly removes large amounts of light chains, improves both renal function and long-term survival expectancies. PMID- 3043078 TI - [Skin expansion in the restorative surgery of burns]. PMID- 3043079 TI - [Effect of cryogenic action on the proteinase activity in patients with keloid and hypertrophic scars]. PMID- 3043080 TI - [Use of immobilized proteases in preparing burn wounds for plastic closure]. PMID- 3043081 TI - [Physical methods of treating burn patients]. PMID- 3043082 TI - [The problem of detoxifying the body today]. PMID- 3043083 TI - [Use of a UHF field to accelerate the epithelialization of donor wounds in burn patients]. PMID- 3043084 TI - [A method for assessing the sorptive capacity of sorbents for microorganisms]. PMID- 3043085 TI - [Immediate and late results of suturing perforated ulcers of the stomach and duodenum]. PMID- 3043087 TI - [Selection of the operation in difficult-to-remove gastric and duodenal ulcer complicated by hemorrhage]. PMID- 3043086 TI - [The complex treatment of acute appendicitis]. PMID- 3043088 TI - [The role of endoscopy in the treatment of ulcer-related gastrointestinal hemorrhage]. PMID- 3043089 TI - [Acute post-traumatic cholecystitis]. PMID- 3043090 TI - [A suture-glue method of treating an appendicular stump]. PMID- 3043091 TI - [Adaptive "figure eight"-shaped knotted removable sutures in obese patients]. PMID- 3043093 TI - [Etiology and pathogenesis of gastritis following operations on the stomach (review of the literature)]. PMID- 3043092 TI - [A method of suturing a bleeding gastric vessel]. PMID- 3043094 TI - [Postoperative peritonitis (review of the literature)]. PMID- 3043095 TI - [Intraoperative hemodynamics during transplantation of a kidney from a living related donor]. PMID- 3043096 TI - [Reaction of tissue to metallic staples]. PMID- 3043097 TI - [Development and current status of silicone oil surgery]. AB - On the basis of a series of 500 patients the authors describe the development of silicone oil surgery, the surgical technique, and present-day indications for it, as well as the results that can be achieved with this procedure. The postoperative problems (cataract, glaucoma, and keratopathy) are analyzed and guidelines presented for reducing the complication rate. PMID- 3043099 TI - [Pilot through 125 years of ophthalmologic science and practice. Rudolf Tiel (1894-1967)]. PMID- 3043098 TI - [Bullous dermatosis with conjunctival and corneal involvement: differential diagnostic scope and explanation of the "overlap syndrome" concept]. AB - Chronic blister-forming dermatosis can lead to conjunctival and corneal involvement. Taking one such case as an example, the authors show that while disorders of this kind can be classified as a form of bullous dermatosis, the differential diagnostic classification is not unequivocal, regardless of the examination method adopted. The term "overlay syndrome" has been introduced into the dermatologic literature to cover clinical pictures of this kind. PMID- 3043101 TI - [Lesions of the posterior visual pathways: clinical presentation and mechanisms of functional restoration]. AB - The present paper begins with a brief discussion of general clinical features found in patients with lesions of the posterior visual pathways. The author goes on to discuss various mechanisms of restitution of function in the central nervous system, including activation of neural pathways subserved by spared neurons, metabolic modulations, and neural plasticity. PMID- 3043100 TI - [Hemianopsias in cortical and subcortical lesions of the visual system: what is the difference?]. AB - The neuronal correlate of perception in the visual area remains unimpaired in lesions of the optic tracts or the radiatio optica. Hemianopsias due to lesions of these pathways are perceived immediately. On the contrary, hemianopsias due to a lesion of the visual cortex are not perceived or are only indirectly realized. It has not been established if a lesion of the primary visual area alone is sufficient to make the patient unaware of a hemianopsia or if additional impairments of the peristriate visual areas are required. Data processing within the cortical visual projection chain makes it likely, however, that a lesion of the striate cortex is enough. Under physiological conditions an uniformly illuminated field is represented in the neuronal activity by the borders of the field. Neurons within the field are not activated. The higher visual cortical areas can so not discriminate if an input missing from V1 is a result of the physiological processing or of a lesion. Scotomas are therefore not perceived. By analogy this argument should also hold for a hemianopic "scotoma". PMID- 3043103 TI - [Irradiation of choroid melanoma: justifiable therapy or medical experiment]. AB - An analysis of published results of irradiation of choroidal melanomas shows that only two reports contain justified statistics. All other statistics are based on insufficiently long follow-up periods, and do not cover all patients irradiated so that artifacts of selection occur which cannot be checked. Arguments are adduced which show that the authors of the two relevant reports mentioned rightly changed their previously positive attitude towards irradiation of these tumors. With the exception of three circumscribed categories of patients, irradiation of choroidal melanomas is still an unjustifiable medical experiment. PMID- 3043104 TI - [Severe hypernatremia in acquired disorder of thirst and vasopressin regulation]. AB - Three patients with severe adipsic hypernatremia (greater than 171 mmol/l) are presented. In two of them, hypernatremia occurred after the operation of a ruptured aneurysm of the A. communicans anterior, in one patient the cause of the disease remained obscure. Despite high plasma osmolality, all patients had low or undetectable plasma vasopressin levels, even throughout hypertonic saline infusion. Urine concentrating ability was partially maintained, suggesting activation of alternative extra- and intrarenal concentrating mechanisms or increased renal sensitivity to low vasopressin concentration. Nonosmolar stimulation (insulin-induced hypoglycemia) did increase vasopressin concentration only subnormally in the two patients tested. This finding might be due to an extended and complex dysfunction of the anterior hypothalamus rather than to a circumscribed defect of the osmoreceptor/thirst center or the supraoptic nuclei. PMID- 3043102 TI - [Homonymous hemianopsia in brain tumors]. AB - Homonymous hemianopsia may be caused by tumors in the region of the optic tract, the lateral geniculate body, the optic radiation, and the visual cortex. Tumors are responsible for about two-thirds of the temporal lesions and about one-half to one-third of the parietal and occipital lesions. With brain tumors a chronological sequence of two groups of signs and symptoms is the rule: first the focal symptoms corresponding to the tumor lesion in a defined cerebral area, later the distant effects of the increasing volume of the tumor, which lead to the general signs of increased intracranial pressure. The different types of homonymous hemianopia in tumor lesions along the suprachiasmatic pathway (optic tract, lateral geniculate body, temporal lobe, parietal lobe, occipital lobe) are described and discussed. The general neurological signs and symptoms are briefly reviewed. Demonstration of important cerebral diagnostic examination methods (plain X-ray, electroencephalogram, computer tomogram, nuclear magnetic resonance, angiography). Differential diagnosis of brain tumors (hematomas, abscesses, granulomas, parasites etc.). PMID- 3043105 TI - New model systems for hepatitis B virus research. PMID- 3043106 TI - Pulmonary removal of circulating endotoxin results in acute lung injury in sheep. AB - Endotoxemia has often been associated with the development of the adult respiratory distress syndrome. Our previous studies have shown that sheep, a popular animal model for adult respiratory distress syndrome, have abundant resident pulmonary intravascular macrophages that rapidly remove inorganic particles and live bacteria from the circulating blood. In this study, we examine the fate of circulating endotoxin in sheep and correlated the site of uptake with early morphologic evidence of tissue injury. Mature sheep and rats received intravenous 125I-labeled lipopolysaccharide (LPS). The dose was 0.8 microgram/kg in sheep and 17.0 micrograms/kg in rats. 125I-LPS clearance from the blood was assayed by gamma-counting of blood samples drawn over 1 hour and was correlated with circulating leukocyte numbers. The distribution of 125I-LPS was determined by gamma-counting of samples of major organs and tissues at time of necropsy. Lungs and liver were examined morphologically in both species. The half-life of circulating 125I-LPS was 2.38 minutes in sheep, and 12.39 minutes in rats. The endotoxin content of the lungs after injection was 77.58% of the total recovered dose in sheep, but only 2.02% in rats. Neutrophil margination occurred in the lungs of both species. In sheep, almost 25 minutes elapsed before peripheral neutrophil numbers decreased by 50%, much longer than the time required for LPS sequestration in the lungs. Rapid LPS uptake by the sheep lungs was associated with early (10-minute) ultrastructural changes including signs of pulmonary intravascular macrophage activation and microvascular congestion. By 60 minutes, many capillaries were occluded with neutrophils, platelets, and fibrin. There was interstitial edema, and endothelial cells showed evidence of severe injury. We conclude that in contrast to the rat, the sheep clears circulating endotoxin more rapidly and preferentially sequesters it in the lungs. Subsequent release of mediators by activated pulmonary intravascular macrophages may then lead to influx of other inflammatory cells and cascading injury. PMID- 3043107 TI - Effect of bacterial sepsis on gluconeogenic capacity in the rat. AB - Since sepsis places increased demands on the host for energy and on other substrates for tissue repair and host defense, hepatic gluconeogenesis is critical for the host's adaptation to sepsis. Substrate-stimulated gluconeogenesis (i.e., gluconeogenic capacity) was assessed by the alanine load method in mannoheptulose-pretreated rats made septic by cecal ligation after laparotomy, as well as by cecal ligation and puncture after laparotomy. Fasted rats subjected to laparotomy only (sham-ligated) and fasted, nonoperated rats (controls) were investigated simultaneously. Following an overnight (-18 to 0 hr) fast, nonoperated animals converted 17.9 +/- 1.5% of [14C]alanine to [14C]glucose. Continued fasting in nonoperated animals resulted in enhanced (P less than 0.05) gluconeogenic capacity (6 hr = 27.2 +/- 3.0%; 24 hr = 26.2 +/- 1.9%; and 48 hr = 28.5 +/- 2.6%) relative to Time 0. Laparotomy alone (sham ligation) delayed the fasting-induced increase (P less than 0.05) in gluconeogenesis capacity (6 hr = 21.1 +/- 1.2%; 24 hr = 18.5 +/- 1.3%; 48 hr = 27.8 +/- 1.0%) relative to Time 0. In contrast, postoperative sepsis produced a sustained depression (P less than 0.05) of gluconeogenic capacity relative to nonoperated sham-ligated controls at 48 hr (cecal ligation, 18.4 +/- 1.4%; and cecal ligation and puncture, 18.8 +/- 1.2%). Thus, (1) fasting enhances hepatic gluconeogenic capacity; (2) surgical trauma transiently blunts the gluconeogenic response to fasting; and (3) sepsis undermines the gluconeogenic response to fasting. PMID- 3043109 TI - Enhancement of growth of aerobic, anaerobic, and facultative bacteria in mixed infections with anaerobic and facultative gram-positive cocci. AB - The potential for mutual enhancement of growth of seven strains of anaerobic and facultative gram-positive cocci (AFGPC), seven aerobic and facultative organisms, and two Bacteroides spp. commonly isolated with AFGPC in mixed infection was evaluated. Enhancement was studied by measuring the relative increase in the colony-forming units of the two bacterial components inducing subcutaneous abscesses in mice. Of the 56 combinations, AFGPC were enhanced in 6 and inhibited in 1. The aerobic and facultative bacteria were enhanced in 32 of the 42 combinations and depressed in none. The Bacteroides spp. were enhanced in 12 of the 14 combinations and suppressed in none. The organisms uniformly enhanced by AFGPC were Group A streptococci, Pseudomonas aeruginosa, and Bacteroides fragilis (all seven instances), followed by Escherichia coli (six of seven instances), Bacteroides asaccharolyticus and Klebsiella pneumoniae (five instances each), Staphylococcus aureus (four instances), and Group D streptococci (three instances). It is apparent that in mixed infection with AFGPC the rate of growth of Bacteroides spp. and facultative and aerobic bacteria is enhanced much more than the rate of growth of AFGPC. PMID- 3043108 TI - Oral administered nonabsorbable antibiotics prevent endotoxemia in primates following intestinal ischemia. AB - Plasma lipopolysaccharide (LPS) concentrations have been found to increase during a temporary occlusion of the superior mesenteric artery (SMA). We have attempted to show, by a prophylactic oral administration of a nonabsorbable antibiotic to monkeys subjected to an SMA occlusion shock, that the increased LPS is intestinal in origin. A total of eight monkeys were subjected to a temporary occlusion of the SMA. Four monkeys received prophylactic oral administration of a nonabsorbable antibiotic, while the rest acted as controls. The plasma LPS concentrations before occlusion in the control and the kanamycin group were 0.069 +/- 0.006 and 0.092 +/- 0.005 ng/ml, respectively. At the end of the 1-hr occlusion period the plasma LPS concentration in the controls increased to 0.09 +/- 0.009 ng/ml (P less than 0.1) and peaked to 0.378 +/- 0.103 ng/ml (P less than .001) within 20 min of reperfusion. Thereafter, the plasma LPS concentration returned slowly to baseline. In the kanamycin group the plasma LPS concentration remained at baseline throughout both the occlusion and reperfusion periods. These data suggest that the origin of the increased plasma LPS concentration seen following temporary occlusion of the SMA is from the gut, and is information of possible importance in patients about to undergo intestinal surgery. PMID- 3043110 TI - A glycolipid precursor of bacterial lipopolysaccharide (lipid X) lacks activity against endothelial cells in vitro and is not toxic in vivo. AB - Lipid X (2,3-diacylglucosamine-1-phosphate) accumulates in mutants of Escherichia coli incapable of assembling the disaccharide backbone of lipid A, the principle endotoxic moiety of bacterial lipopolysaccharide (LPS). We compared the effects of lipid X on cultured bovine aortic endothelial cell (BEC) viability and prostacyclin (PGI2) release with those of lipid A and LPS. At 10(-5) M, both LPS and lipid A produced significant BEC cytotoxicity (percentage cytotoxicity 69 +/- 4 for LPS and 51 +/- 11 for lipid A) and induced a variable but consistent increase in the release of PGI2 (11- to 73-fold increase for LPS and 4- to 6-fold increase for lipid A). Lipid X, in contrast, was not toxic and did not induce PGI2 release at 10(-4) M. Pretreatment and coincubation of BEC with lipid X, at a concentration 100 times greater than LPS, failed to prevent LPS-mediated cytotoxicity. Intravenous infusion of lipid X in goats had no effect except for a modest elevation in the pulmonary artery pressure during the period of infusion. Moreover, pretreatment of goats with lipid X (70 micrograms/kg) did not block the effects of a subsequent infusion of LPS (5 micrograms/kg). These data suggest that a fatty acid-substituted disaccharide is the minimal molecular requirement for the numerous effects in vivo and activity in vitro induced by LPS. Furthermore, these effects are not prevented by pretreatment with a monosaccharide precursor of lipopolysaccharide, lipid X, at a dose 10- to 100 fold greater than that of LPS. PMID- 3043111 TI - A new nitrosourea derivative for the treatment of chronic myelogenous leukemia. AB - A new water-soluble nitrosourea derivative, methyl 6-[3-(2-chloroethyl)-3 nitrosoureido]-6-deoxy-alpha-D-glucopyranoside (MCNU), was found to be useful for the treatment of chronic myelogenous leukemia (CML) in the chronic phase. To compare the efficacy of MCNU with that of busulfan, patients were randomized. In the 40 patients administered MCNU, the median time to the achievement of a complete remission (CR) was 50 days. This value was shorter than that observed in 37 patients administered busulfan (126 days, p less than 0.05). There were no differences in the rate of CR achieved, mortality, median time to the onset of blast crisis (BC), BC rate, or survival rate during the observation period. The overall incidence of side effects was higher for MCNU (31%) than for busulfan (15%), but the symptoms were mild, transient and tolerable for most patients. These results suggest that MCNU is a safe and valuable addition to the therapeutic repertoire for the control of CML. PMID- 3043112 TI - A simple modular tissue bath developed for 'in vitro' studies of the isolated spinal cord. AB - A tissue bath for studies of the mammalian spinal cord in vitro is described. The modular construction of the bath permits considerable flexibility for accommodating different tissue preparations. The individual components of the bath are easily constructed and the design permits multiple compartment baths to be simply formed. The design principle on which the bath is based may have much wider application than that described. PMID- 3043113 TI - A low-priced computer-assisted densitometer and analysis software for quantitative autoradiography. AB - A computer-assisted densitometer which consists of a darkroom enlarger, a black and-white exposure meter, an amplifier, an analog-to-digital converter, and a microcomputer with a monitor and a graphic printer can be utilized for the quantitative analysis of autoradiograms. QUANTAR, a computer program written in BASIC, records the density measurements and stores the data in a file that can be easily retrieved by commercially available spreadsheet software. A spreadsheet template was designed to convert the digital readings into concentration of ligand in tissue. A variety of spreadsheet templates can be created to analyze data for a standard curve, a Scatchard plot, or a binding competition curve. This system, which is easy to assemble, may be useful to the frugal investigator with a modest equipment budget. PMID- 3043114 TI - Recombinant interleukin-2 limits the replication of Mycobacterium lepraemurium and Mycobacterium bovis BCG in mice. AB - BALB/c mice were infected with Mycobacterium lepraemurium (MLM) in the foot pad or with M. bovis BCG intravenously with 5 x 10(7) bacilli. Recombinant interleukin-2 (IL-2) was injected intraperitoneally as a single dose (20,000 u), single course of 5 injections (400 u each) or 6 monthly courses starting 3 days or 60 days after the MLM infection. BCG infected mice received a single dose (1000 u) or 5 daily injections of 100 or 1000 u each. IL-2 significantly reduced the total bacterial counts in the footpad, lymph node and liver of MLM infected mice (50-85%) by 6 months and viable counts in the spleen (30-50%) by 60 days after BCG infection. The courses of IL-2 started at 60 days were more effective than at 3 days after MLM infection (P less than 0.05-0.001) and in the case of BCG, 100 u of IL-2 was better than 1000 u (P less than 0.05-0.01). These results indicate that IL-2 limits mycobacterial infections in mice, and raise the question of its possible use in humans. PMID- 3043115 TI - Towards the morphology of medical information systems. AB - A review of research and development in the field of medical information systems shows that, on the whole, technical hardware and specific software problems have been the central consideration. Only recently has it been acknowledged that it is necessary to include the formerly neglected specific functional and organizational aspects. In this respect, morphological approaches enable new perspectives to be opened up for the analysis, design and classification of medical information systems. PMID- 3043116 TI - Cognition activators. PMID- 3043117 TI - Synthesis and in vitro activity of the penem antibiotics. PMID- 3043118 TI - [Ultrasonic diagnosis of the salivary glands]. AB - Sonography of the salivary glands is a valuable diagnostic tool in acute and chronic sialadenitis, sialadenoses, lymphadenopathies and tumours lesions. A new examination technique is presented: ultrasound-guided fine needle biopsy. We report on our experience in 308 patients with parotid and in 47 patients with submaxillary gland diseases examined between 1982 and 1987. The diagnostic accuracy and limitations of sonography are discussed compared with sialography, computed tomography and magnetic resonance imaging. PMID- 3043120 TI - [Angiodynography: a new imaging procedure in the ENT field]. AB - A new diagnostic imaging is presented which demonstrates the blood flow in addition to the usual ultrasonic image. Motion pictures show both the direction of the blood flow in different colours and the speed in variable intensity of the colour. Thus vessels of only 0.5 mm diameter may be identified. In otorhinolaryngology, the vascular supply of space-occupying lesions can be determined quantitatively in a noninvasive way. Lymphomas may be differentiated from vascular tumours and malformations. Preoperatively, vessels adjacent to the tumour may be identified. This method is particularly useful in the evaluation of thyroid and salivary gland disorders. PMID- 3043119 TI - [Differential echographic diagnosis of salivary gland tumors]. AB - 76 patients with tumorous swellings of the salivary glands were examined by means of standardized A-scan and B-scan sonography. B-scan echography allowed a differentiation between benign and malignant tumours in all cases. Analysis of the various A-scan criteria (internal structure, reflectivity, borders, consistency and sound attenuation) provided a pathognomonic combination of these criteria for each lesion, enabling further histological diagnosis. PMID- 3043121 TI - [Arterial and venous digital subtraction angiography. A current study technic for otorhinolaryngology]. AB - Digital subtraction angiography (DSA) is a rather new radiographic technique for imaging different vascular alterations, particularly in the head and neck area, and can replace angiography in most cases. Nevertheless, routine application of DSA - intravenously or intraarterially - seems to present a problem to many clinicists. In the present paper, therefore, we try to introduce these important techniques by means of several examples from patients who have undergone these procedures to demonstrate the actual indications of DSA for oto-rhino laryngological diseases. PMID- 3043122 TI - [Reconstruction of the lower third of the face--microsurgical monobloc transfer with vascularized bone]. AB - The authors present two microsurgical options of the reconstruction of the lower third of the face. They outline a systematisation of indications and techniques and emphasise versatility and flexibility of free flaps compared to pedicled flaps, and describe the advantages of the vascularisation and the techniques of the iliac crest osteocutaneous flap and the parascapular cutaneous-latissimus dorsi osteomyocutaneous flap which was developed by Nassif. PMID- 3043123 TI - [The history of tracheotomy]. AB - Tracheotomy is one of the oldest operations in medicine. The development of tracheotomy from antiquity to our time is reported. After the beginnings of the operation in Greek and Roman medicine, the operation was called laryngotomy or bronchotomy. It was Heister who coined the name "tracheotomy". Tracheotomy is still an important operation even after more than 2000 years. In recent years there has been a change from simple tracheotomy to plastic tracheostomy. PMID- 3043124 TI - [Morphometry of the larynx in horizontal sections. Normal data for the quantitative evaluation of current imaging technics]. AB - Laryngeal anomalies as a predisposing factor for pathogenesis of vocal disturbances have been discussed for a long time. Laryngeal structures can be made visible in vivo, and structural details be measured, only since the development of new imaging techniques, mainly of CT. As adequate descriptions have not been published in literature unto now, we measured horizontal sections of laryngeal specimens to collect reference data for a quantitative analysis of CT images. An evaluation technique was therefore developed, which, by clear definition of the planes and points of measurements, yields reproducible results. Direct measurement of the thyroid angles can be abandoned, since these can be easily calculated from the known distances. PMID- 3043125 TI - [Auxiliary motor innervation of the laryngeal muscles via the internal branch of the superior laryngeal nerve]. AB - According to our present knowledge of the neuromuscular innervation of the intrinsic laryngeal muscles, the cricothyroid muscle is innervated by the external branch of the superior laryngeal nerve (NLS), whereas all other remaining muscles get their supply from the inferior laryngeal (recurrent) nerve. Mainly in the phoniatric literature, however, opinions differ concerning an additional motoric laryngeal innervation. In human larynges, excised for large unilateral carcinoma, horseradish peroxidase (HRP) was injected into the internal branch of the NLS. Anterograde labelling of axons was demonstrated histochemically. In adjacent sections of the different muscles, end plates and axons were stained histochemically with silver impregnation and acetylcholinesterase. Evidence is presented of motor innervation of the internal branch of the NLS in some laryngeal muscles. With retrograde HRP-tracing in sheep, motoneurons were detected in the nucleus ambiguus, although the recurrent nerve and the external branch had been divided and excised. Thus, histologically an additional neuromuscular supply via the internal branch of the NLS is demonstrated. PMID- 3043126 TI - [Congenital cysts of the larynx]. AB - Based on our own observation of one case of congenital laryngeal cyst of the newborn and a review of 48 cases presented in literature, this congenital anomaly is described. The clinical picture is demonstrated and the genesis of these cysts is discussed. PMID- 3043127 TI - [Sonography of the pancreas]. AB - Sonography was the first non-invasive method that allowed reproducible demonstration of the pancreas. The organ is located in the anterior pararenal space, it is 12-15 cm long. The pancreatic head is in direct contact with the duodenum, the body is located behind the posterial wall of the stomach and the pancreatic tail has topographic relation to the left kidney and to the hilum of the spleen. The main guiding vessel in the transverse scan is the splenic vein. The main value of sonography in acute pancreatitis is description of the course of the inflammatory process and delineation of a biliary etiology. The sensitivity in the demonstration of pseudocysts larger than 2-3 cm is about 90%. In chronic pancreatitis sonography often demonstrates a normal organ. In advanced cases an irregular echopattern with echogenic zones and irregular outer contours are seen. Endoscopic sonography is particularly valuable in demonstrating abnormalities of the pancreatic duct. Dilatation of its tributaries and irregularities of its wall are sharply depicted. 80% of pancreatic carcinomas are echopoor and carcinomas between 2-3 cm in size can be diagnosed by sonography in a few patients. However, the overall prognosis of pancreatic carcinoma has remained unchanged in the last years. PMID- 3043128 TI - [Treatment of duodenal and prepyloric ulcers with an antacid and cimetidine either alone or in combination]. AB - In an open randomized study the effectiveness and the acceptance of treatment with a cytoprotective antacidum (226 mval acid-neutralisation-capacity per day's dose) have been examined in 60 patients with clinical and endoscopical secured prepyloric and duodenal ulcera compared with Cimetidine (2x 400 mg) and an initial combination in the first week of treatment. The healing rates depend clearly on the size of ulcus at the beginning of treatment. Ulcera smaller than 8 mm heal in 3 1/2 to 4 weeks up to 71% with the Antacidum, up to 56% to 83% with Cimetidine. For larger ulcera the healing rates are clearly poorer (antacida 20%, Cimetidine 33-67%). Initial combination treatment of the antacidum with Cimetidine for 1 week and the further treatment with the antacidum shows a healing rate of 100% for ulcera smaller than 8 mm, for larger ones a rate of 75%. Therefore the initial combination treatment seems to be useful especially for ulcera duodeni and prepyloric ulcera larger than 8 mm. PMID- 3043130 TI - [Hemangioma of the liver: often an incidental finding]. AB - Hemangioma is the most frequent benign liver tumor with an incidence of 0.4-7.3%. The tumors are typically small, asymptomatic and detected accidentally by ultrasound or CT. Hemangiomas extending a diameter of 2-3 cm can be proved by dynamic contrast CT or scintigraphy with 99-m Tc erythrocytes. Hemangiomas with a diameter less than 2 cm are a diagnostic dilemma. In case of a typical ultrasonic picture (small white tumor) in an asymptomatic patient, a consequent ultrasonic follow-up is indicated. It is yet unclear whether MRI will be an additional diagnostic tool to identify small hemangiomas. PMID- 3043129 TI - [Chemotherapy of advanced pancreatic cancer with 5-fluorouracil, doxorubicin and high-dose methotrexate]. AB - The aim of the present phase II clinical trial was to investigate the therapeutic efficacy and tolerance of a combination chemotherapeutic protocol consisting of 4 weekly intervals of 5-fluorouracil, doxorubicin and high-dose methotrexate (FAMeth-regimen) in patients with advanced measurable pancreatic cancer. After a median treatment duration of 4 (2-12) months, one complete and one partial response were achieved in the 13 evaluable patients. Two additional patients had evidence of objective tumour regression, although response was less than 50% of pretreatment tumor measurements. Stable disease was noted in 3 patients, and the tumour progressed in 6. The median survival of all evaluable patients from start of therapy is 7 (2-17) months. Side-effects associated with FAMeth-chemotherapy were generally mild and reversible and primarily included gastrointestinal symptoms (38%) and leukopenia (62%). There was 1 treatment related death due to pancytopenia and sepsis. Our preliminary data suggest some antitumour activity of the regimen against pancreatic cancer, although final assessment of therapeutic results must await accrural of additional patients. PMID- 3043131 TI - Membrane effects of ethanol: bulk lipid versus lipid domains. AB - It has been well-established that ethanol fluidizes the bulk lipid of membranes and that this effect may alter cell function and be involved in ethanol sensitivity and tolerance. This hypothesis has been supported in several studies, however, there is also a considerable amount of data that do not support such an explanation, e.g., direct effect of ethanol on proteins, other membrane acting drugs, temperature effects, effects of ethanol on aged membranes and inconsistent effects of chronic ethanol consumption on lipid content. This review examined the bulk membrane fluidization hypothesis in light of those data and proposed a modification of the bulk membrane hypothesis that is based on recent data that show that ethanol and other alcohols have a specific effect on the structural properties of different membrane domains. This specific effect of ethanol is discussed within the context of how changes in fluidity of domains may alter membrane function. PMID- 3043132 TI - Behavioural effects of N-methyl-D-aspartate in the anterodorsal striatum of the rat. AB - N-methyl-D-aspartate (NMDA) (0.5 and 1 microgram/0.5 microliter) bilaterally injected into the anterodorsal striatum of rats reduced locomotion, sniffing, rearing and feeding upon presentation of palatable food. Consequently, the number of all behavioural bouts exhibited was reduced and the duration of akinetic phases was prolonged. These results are discussed in connection with previous findings showing that the NMDA receptor blocker DL-2-amino-5-phosphonovaleric acid (AP-5) injected at the same site - produced opposite effects: AP-5 enhanced locomotion, rearing, sniffing as well as the total number of behavioural bouts exhibited. PMID- 3043134 TI - [An integrated computerized anesthesia record: indexing with a terminology dictionary]. PMID- 3043133 TI - [Postoperative pain relief with various epidural narcotics: demerol, butorphanol, nalbuphine, and morphine]. PMID- 3043135 TI - [Clinical application of pulse oximeter]. PMID- 3043136 TI - [Reliability of clinical studies in the diagnosis of malaria fever in West African endemic areas]. AB - We have studied 930 febrile cases in a savanna area of Burkina Faso. We have diagnosed malarial fevers according to the only clinical data in 333 cases and it was confirmed in 154 cases by parasitological study; for 597 cases a non malaria reason was given according to the only clinical data and it was confirmed in 507 cases by biological studies. On the whole the error rate was of 28% on the clinical examination alone and this proportion was equal or superior whatever the parasitological threshold was. The clinical examination is not allow by it self to diagnose a malaria fever case. PMID- 3043137 TI - [Malaria in the Republic of Djibouti. Strategy for control using a biological antilarval campaign: indigenous larvivorous fishes (Aphanius dispar) and bacterial toxins]. AB - The authors take stock of the present situation of malaria in the Republic of Djibouti which, after several decades of silence, seems to have been reintroduced at the beginning of the seventies. Actually it is hypo-endemic malaria with Plasmodium falciparum of which the only vector seems to be Anopheles arabiensis, gambiae complex. The specificity of the larvae nests allows a control strategy based on the only treatment of larvae sites by biological control: larvivorous fishes (Aphanius Dispar) and in addition the pin-point use of bacterial toxins as a complementary measure. The first results obtained in the rural zones around the capital are encouraging and permit to envisage the extensions of such a strategy to the whole of the territory of the Republic. PMID- 3043138 TI - [Current strategies in the campaign against Plasmodium falciparum malaria in an area resistant to 4-aminoquinolines]. AB - Confronted with the extension of chemoresistance of P. falciparum and the gradual renunciation of the fight against the vectors, it is important nowadays to manage better the means of fighting in the zones where chloroquine resistance exists. If preventive treatment of chills by 4-aminoquinolines is still able to realize prevention of mortality, treatment of all confirmed chills by P. falciparum has to combine sulfadoxine-pyrimethamine with quinine or amodiaquine. PMID- 3043139 TI - [Response of Plasmodium falciparum to quinine in a hospital environment in a chloroquine-resistant region. Bujumbura, Republic of Burundi]. AB - Plasmodium falciparum's response to quinine (7.5 g of base in 5 days) was studied in vivo in 58 adult patients hospitalized in 1985--47% presented a pernicious malaria fever three deaths were registered during treatment, two of them attributed to late hospitalization, and the third patient was struck down by a sudden acute hepatitis. Three days after treatment started 89 of the patients did not present any asexual forms. At day 7, all patients responded favourably to the treatment. Parasitemia lowered very quickly whatever the route of administration had been. Quinine had no effect on production of gametocytes during the first four days. Clinical improvement is rapidly observed and the side effects of quinine disappeared when stopping the treatment. Despite the presence in the region of an important resistance of Plasmodium falciparum to chloroquine (80% specimens), one can conclude to a satisfying susceptibility of this parasite to quinine, provided posology and divided doses are respected. PMID- 3043140 TI - [The role of surgery in acute epididymitis]. AB - One third of the acute epididymitis (A.E.) admitted in the Military Hospital in Marseilles have been operated, i.e. 33 cases; out of them the last 12 cases have been systematically monitored by ultrasonography. Twice out of three times, clinical picture suggested a possible twisting that led to an emergency surgical exploration. Interest of ultrasonography of the scrotum is underlined as far as diagnosis is concerned but also in monitoring A.E. Indeed, despite medical treatment (antibiotics + anti-inflammatory drugs), it is possible that some A.E. are evolving either to abscess or to their "vascular" form; The ultimate form of the acute epididymo-orchitis is necrosis of the testis. This is due to, or amplified by, vascular compression linked up with edema. Surgical intervention has therefore to investigate by inguinal path the entire spermatic cord and the epididymis-testis system, and to apply the "decompression technique" requested. PMID- 3043141 TI - Widespread distribution of deletions of the bgl operon in natural isolates of Escherichia coli. AB - A deletion that includes the bgl (beta-glucoside utilization) operon of Escherichia coli was originally detected in several rarely occurring natural isolates that utilize cellobiose. Here I show that bgl deletions are present in 95% of the Cel+ isolates obtained from diverse sources. They are also present in 29% of the Cel- strains in two different collections of natural isolates of E. coli. At least three versions of bgl deletions are present in E. coli populations. In the most common version approximately 8 kb of DNA around the bgl region of E. coli K12 is replaced by a specific 6.5-kb DNA fragment. In another version a deletion of similar length is not replaced by the same sequence. A third version involves deletion of approximately 14 kb without the replacement fragment being present. The distribution of these deletions suggests that the version 1 deletion occurred very early in the history of E coli. It also appears likely that there is selection for bgl deletions in Cel+ strains of E. coli. The presence of the version 1 deletion within distantly related phylogenetic groups of E. coli provides evidence for recombination within natural populations of E coli. PMID- 3043142 TI - [Genetic problems in occupational medicine]. AB - The investigation has been aimed at the genetically conditioned increased sensitivity to occupational hazards. Special attention should be paid to population's genetic differentiation at setting up hygienic standards and interpretation of results of biological absorption tests. As problems related to genetically conditioned increased sensitivity to hazards are particularly significant in the countries with the population's considerable biochemical differentiation or considerable rate of the so called biochemical deficits, therefore the problem gets more intense as more immigrants from Asiatic and African countries are employed in European industries. In the authors' opinion, correctly arranged pre-employment studies should reduce the number of poisonings in chemical industry. PMID- 3043143 TI - Relationship between changes in glucose production and gluconeogenesis during mild hypoglycemia in humans. AB - We measured 14C-alanine conversion to 14C-glucose (an index of gluconeogenesis) and glucose production in six healthy volunteers during low-dose insulin infusion (0.3 mU/kg.min for four hours). Insulin rose from 7 +/- 2 to 20 +/- 2 microU/mL, and plasma glucose fell to a plateau of 65 to 70 mg/dL after 60 minutes. Glucagon and catecholamines increased after 60 minutes, whereas C-peptide decreased immediately. Glucose production decreased transiently by 43% and then returned to baseline after 45 minutes. In contrast, 14C-alanine conversion to 14C-glucose was unchanged for 120 minutes, but then rose twofold above baseline by 240 minutes. Our data suggest that early recovery of glucose production during mild hyperinsulinemia occurs independent of changes in gluconeogenesis. However, gluconeogenesis plays an increasingly more important role in maintaining glucose production when mild hypoglycemia is prolonged. PMID- 3043145 TI - Decreased response to catecholamines in the newborn: effect on glucose kinetics in the lamb. AB - Epinephrine, a catecholamine with both alpha (alpha) and beta (beta) adrenergic effects, may produce hyperglycemia in adults by increasing glucose production and decreasing glucose clearance. To document the degree of sensitivity and characterize maturation of neonatal glucose control, glucose kinetics were measured in 14 term newborn lambs (weight 4.5 +/- 0.3 kg [mean +/- SEM] and aged 4.1 +/- 0.4 days). Following infusion of 0.9% NaCl at 0.06 mg.kg-1 min-1 plus 100 microCi/kg D[6-3H] glucose by prime plus constant infusion, rate of production (Ra) and glucose clearance were measured during administration of epinephrine. The responses in the newborns were compared with those in six adult sheep. Under conditions of epinephrine infusion, the plasma glucose concentration in the newborn lambs increased to 129 +/- 18 mg/dL (50 ng.kg-1 min-1 epinephrine), P less than .0001, and 253 +/- 8 mg/dL (500 ng.kg-1 min-1 epinephrine), P less than .0001, compared with 95 +/- 8 mg/dL (controls, no epinephrine). Comparable values for Ra were 6.5 +/- 1.6 mg.kg-1 min-1 (50 ng.kg-1 min-1 epinephrine), P = NS, and 18.5 +/- 3.0 mg.kg-1 min-1 (500 ng.kg-1 min-1 epinephrine), P less than .0001, compared with 5.3 +/- 0.5 mg.kg-1 min-1 (controls, no epinephrine).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3043144 TI - Actions of insulin and insulinlike growth factors I and II in cultured microvessel endothelial cells from bovine adipose tissue. AB - Endothelial cells were cultured from bovine adipose microvessels, pulmonary arteries, and aortas. The effects of insulin, IGF-I, and IGF-II (MSA) on glucose uptake, neutral amino acid (AIB) uptake, and thymidine incorporation into DNA by the endothelial cells were determined. Each hormone markedly stimulated all three processes in the microvessel endothelial cells but had no effect on the larger vessel endothelial cells. In the microvessel cells the monoclonal antiinsulin receptor antibody, Ab 47-9, was observed to specifically inhibit insulin binding in the bovine microvessel cells without having intrinsic activity on the three biologic processes that were stimulated by insulin and the IGFs. When insulin binding was first inhibited by Ab 47-9, dose-response curves for insulin were markedly shifted to the right for glucose uptake, AIB uptake, and thymidine incorporation into DNA. Similar antibody treatment had no effect on dose-response curves of IGF stimulation of any of the three processes. These data further extend the biologic actions of insulin and the IGFs in cultured microvessel endothelial cells. They also suggest that certain functions mutually stimulated by both insulin and the IGFs, ie, glucose uptake, AIB uptake, and thymidine incorporation into DNA, are substantially mediated through homologous receptors. PMID- 3043146 TI - Bed-rest-induced insulin resistance occurs primarily in muscle. AB - Treatment of trauma victims and patients with severe illness may contribute to their metabolic derangements by severely restricting physical activity. We sought to quantitate the impact of absolute bed rest alone on insulin regulation of glucose metabolism in six healthy subjects. Six to seven days of strict bed rest resulted in moderate deterioration in oral glucose tolerance and increased both fasting plasma insulin concentration and the insulin response to an oral glucose challenge by more than 40%. Euglycemic insulin clamp studies demonstrated the development of resistance to insulin's stimulation of whole-body glucose utilization. This change was characterized by a rightward shift of the insulin dose-response curve (insulin concentration at which 50% of maximal stimulation occurred was 45 +/- 3 (SE) microU/mL in the base line period and 78 +/- 8 microU/mL after seven days of bed rest, P less than .01) with little alteration in the maximal response in the rate of glucose uptake (baseline 15.4 +/- 1.4 mg/kg.min and bed rest 14.0 +/- 1.3 mg/kg.min). In contrast to the shift of sensitivity of whole-body glucose utilization to insulin, suppression of hepatic glucose output by insulin was unchanged by seven days of bed rest. Insulin binding to circulating mononuclear cells was not changed by bed rest. These studies demonstrate that the limited physical activity dictated by bed rest for as little as seven days is associated with substantial resistance to insulin's effects on glucose metabolism. Further, the data suggest that these effects occur primarily in skeletal muscle with little change in insulin action on the liver. PMID- 3043147 TI - A synthetic medium for Saccharomyces cerevisiae. AB - Growth of Saccharomyces cerevisiae strain Castelli 20 has been observed in a minimal synthetic medium using different glucose concentrations and without biotin, during the first 30 hrs of its development at 28 degrees C. The yeast's growth was observed spectrophotometrically at 675 nm reading the Optical Density every hour. The minimal medium of Sheperd et al. (1980), with glucose 15 g/L and biotin was modified: the vitamin was eliminated and the concentration of glucose was gradually reduced down to 0.5 g/L. At 5 g/L of glucose concentration and without vitamins the strain grew very well. From our results up to 30 hrs biotin has no influence on the yeast's growth. The medium would be useful to study Saccharomyces cerevisiae physiology during the early period of its development. PMID- 3043148 TI - Computer-assisted learning in British medical schools. AB - A questionnaire survey of British medical schools requested information about school policy and resources and about departmental use of programs and computing hardware. Few schools were sufficiently involved in computer-assisted learning (CAL) to have a clear policy, nor were resources generally available for CAL development. However, at departmental level there was active development in many schools, particularly in preclinical departments. Hardware preference was for BBC micros, but IBM compatibles appear to be growing in popularity. Because of the huge development costs of CAL, collaboration between schools is seen as essential while financial resources are scarce. PMID- 3043149 TI - Collation of student results in practical class experiments in physiology, using a BBC ECONET computer network. AB - Funding was provided to Queen's University by the Department of Education and by the Industrial Development Board for Northern Ireland to provide microcomputers for undergraduate use. An allocation from the grant to the Department of Physiology enabled the purchase of 20 BBC Master 128 microcomputers with monitors used as student work-stations connected together by an ECONET network with a file server using a dual floppy disc drive, two printer servers and two demonstrator stations. A BASIC program was written to analyse the students' practical class measurements which they entered manually at their work-station keyboards. Class results were presented to the students in the form of frequency distribution histograms or X/Y graphs. Program modifications to suit different practicals can be made relatively easily. The time taken to analyse data has been shortened. It is easy for the students to get immediate comparison of their own results with those of the rest of the class--particularly advantageous if the student's own experiment did not work. The class can be divided into groups to study different variations of the experiment and provide the data from each group to the whole class. The students' opinions on whether the equipment had (1) improved the teaching of physiology and (2) provided helpful preparation for the use of computers in medical practice were assessed by a questionnaire which showed that a clear majority felt these aims had been fulfilled. PMID- 3043150 TI - Coeliac disease: an analysis of aetiological possibilities and re-evaluation of the enzymopathic hypothesis. AB - Although it is well established that components of wheat gluten and structurally related cereal proteins produce intestinal damage in Coeliac disease (CD), the primary defect which confers upon the host susceptibility to these dietary substances remains a mystery. To date, three main hypotheses have been framed to explain this susceptibility, but none has yet been proven. Diagrammatic representation of these aetiologic hypotheses facilitates their analysis and illustrates the potential importance of a relatively ignored possibility, i.e., that there may be a defect in the intraluminal phase of the in vivo processing of dietary gluten. We suggest that future work should be directed towards investigating the potential role of abnormalities of this phase in the aetiology of CD. PMID- 3043151 TI - Therapeutic effects of organic germanium. AB - Germanium is present in all living plant and animal matter in micro-trace quantities. Its therapeutic attributes include immuno-enhancement, oxygen enrichment, free radical scavenging, analgesia and heavy metal detoxification. Toxicological studies document Germanium's rapid absorption and elimination from the body, and its safety. Clinical trials and use in private practices for more than a decade have demonstrated Germanium's efficacy in treating a wide range of serious afflictions, including cancer, arthritis and senile osteoporosis. Germanium's anti-viral and immunological properties, including the induction of interferon, macrophages, T-suppressor cells and augmentation of natural killer cell activity, suggest its possible efficacy in treating and/or preventing AIDS. PMID- 3043152 TI - [Diabetes mellitus and the islands of Langerhans]. PMID- 3043153 TI - Ursodiol for dissolving cholesterol gallstones. PMID- 3043156 TI - [Course of septic peritonitis in patients with and without a spleen]. PMID- 3043154 TI - Two algorithms for the three-dimensional reconstruction of tomograms. AB - Three-dimensional (3-D) surface reconstructions provide a method to view complex anatomy contained in a set of computed tomography (CT), magnetic resonance imaging (MRI), or single photon emission computed tomography tomograms. Existing methods of 3-D display generate images based on the distance from an imaginary observation point to a patch on the surface and on the surface normal of the patch. We believe that the normalized gradient of the original values in the CT or MRI tomograms provides a better estimate for the surface normal and hence results in higher quality 3-D images. Then two algorithms that generate 3-D surface models are presented. The new methods use polygon and point primitives to interface with computer-aided design equipment. Finally, several 3-D images of both bony and soft tissue show the skull, spine, internal air cavities of the head and abdomen, and the abdominal aorta in detail. PMID- 3043155 TI - Verification of total body photon irradiation dosimetry techniques. AB - A method of verifying the dosimetry of patients undergoing total body irradiation (TBI) with photon beams having energies from cobalt-60 to 25 MV is presented. A simple set of spot checks at the TBI axis has been used to verify data used for TBI dosimetry. Calculations to verify dose delivered to TBI patients are done in the same manner as those irradiated at standard treatment distances. A simple method of effective field size determination for various anatomical locations in a typical adult is presented. Measurements in an Alderson phantom with thermoluminescent dosimeters and an ion chamber at several anatomical locations indicate that this calculational method can predict the dose along the patient axis to within 4% for 60Co and 18-MV photon beams, provided the dosimetry data are appropriate (as determined by the spot checks). Results of intercomparisons of TBI beam calibration, off-axis and depth-dose data at various institutions visited by the Radiological Physics Center are also presented. PMID- 3043157 TI - [Clinical aspects of magnesium metabolism]. PMID- 3043158 TI - [Ethambutol side effects on the eye]. PMID- 3043160 TI - [Effect of bismuth subsalicylate versus cimetidine on Campylobacter pylori, ulcer healing and rate of recurrence]. PMID- 3043159 TI - [Metabolic control in labile type I diabetes with conventional insulin therapy, basal bolus insulin therapy using pens and continuous subcutaneous insulin infusion with the pump]. PMID- 3043162 TI - [Asbestosis and the lung. I]. PMID- 3043163 TI - AIDS resources. PMID- 3043161 TI - [Late cardiorespiratory sequelae following chemo- and radiotherapy]. PMID- 3043165 TI - [Recent trends in plastic surgery]. PMID- 3043164 TI - A comparison of fentanyl and morphine use in neonates (continuing education credit). PMID- 3043167 TI - [Evaluation of the clinical usefulness of ultrasonography in making a preoperative diagnosis of thyroid carcinoma]. AB - Preoperative ultrasonographic findings of thyroid carcinoma were analysed in terms of their correlation with pathological features on the cut surface of surgically removed specimens, and the results were found to facilitate correct diagnosis. For this study, 414 patients with thyroid tumors, consisting of 172 with carcinomas and 242 with benign tumors, were examined by a 7.5 MHz high resolutional real-time ultrasonography. Sonographic criteria for diagnosing thyroid carcinoma were newly proposed in this study. They were divided into major and minor criteria: The former were findings frequently observed in carcinoma, and the latter were findings observed in benign tumors but, rarely, in carcinoma also. By applying these criteria in thyroid patients, papillary carcinoma was diagnosed more correctly than follicular carcinoma, and solid carcinoma was more easily diagnosed than cystic carcinoma. Microcarcinomas, less than 10mm in maximum diameter, were frequently diagnosed by ultrasonography; 20 out of 36 were detected by these criteria. Intraglandular dissemination was also detected in 22 out of 87. From these studies, it was clear that the accuracy of ultrasonographic diagnosis for microcarcinoma and intraglandular dissemination was closely related to the size of the tumor. The diagnostic limitation was a maximum tumor diameter of 5mm. The superiority of ultrasonography to other diagnostic methods was also discussed. PMID- 3043166 TI - [Experimental study on glucose turnover and amino acid metabolism after total pancreatectomy]. AB - Glucose turnover and plasma amino acid profile were measured in total pancreatectomized dogs to clarify the mechanism of hyperaminoacidemia after total pancreatectomy. Twenty-one male mongrel dogs underwent total gastrectomy and pancreatectomy and were divided into two groups. I(-) group (n = 11) was treated without insulin and I(+) group (n = 10) received insulin postoperatively. Another 15 dogs of S group received only splenectomy. Glucose appearance rate (Ra), plasma amino acid profile and blood sugar level (BS) were measured before operation, and on the 1st and 3rd postoperative days. Significantly higher Ra and BS were observed in I(-) group than in S group postoperatively and either decreased or unaltered by the glucagon infusion. On the other hand, Ra and BS in I(+) group were maintained at the same levels as S group and increased significantly by the glucagon infusion. Hyperaminoacidemia was observed after total pancreatectomy regardless of insulin treatment. Branched chain amino acids (BCAA) were significantly elevated in I(-) group but not in I(+) group. Glucagon infusion decreased almost all amino acids levels except BCAA in both groups. These results suggest that mechanism of hyperaminoacidemia observed in totally pancreatectomized dogs is the reduced amino acids uptake by the liver related to glucagon deficiency. PMID- 3043169 TI - [Echographic diagnosis for deep vein thrombosis of the lower extremities]. AB - Echographic examination of the femoral vein was carried out in two positions: first in supine position and then in standing position, in order to make a diagnosis of deep vein thrombosis (DVT) of the lower extremities. The distance between the wall images of the femoral vein was measured on the line passing through the center of the femoral artery and crossing the common tangent line of the femoral artery and vein at right angle, and the ratio of this distance in supine position to that in standing position was named FEMORAL VEIN DISTENSIBILITY INDEX (FVDI). The FVDI was 2.83 +/- 0.83 in control group consisting of 14 healthy subjects, and 1.26 +/- 0.18 in DVT group consisting of 21 patients with 23 legs. There was a statistically significant difference between the two groups (p less than 0.001). No statistical difference of the FVDI was, however, noted between the control group and the diseased group consisting of 19 contralateral, symptom-free legs. And no relation was confirmed between FVDI and venographical findings, namely site and extension of thrombotic occlusion. By introducing the FVDI, which reflects functional hemodynamics of the femoral vein, the echographic diagnosis for DVT has become easier and more accurate, presenting a diagnostic accuracy of 90.6% in this study. PMID- 3043170 TI - [Nonpenetrating brachiocephalic arterial injury]. AB - The injury of brachiocephalic artery is uncommon in the blunt trauma and sometimes it was accompanied by other organ damages. We reported a case and reviewed 47 cases in literature. A 28-year-old woman was transferred to our hospital with the blunt chest trauma and the cerebral infarction due to the traffic accident. The cineangiogram showed complete obstruction at the middle portion of brachiocephalic artery and the subclavian steal-carotid recovery phenomenon. Thirty days after trauma, the operation was performed under the monitoring of right superficial temporal arterial pressure. The vessel with intimal defect was plugged by a clot and it was replaced with a Gore-Tex graft during simple occlusion of the right carotid, subclavian and brachiocephalic arteries without any monitoring pressure change. The postoperative course was uneventful and the symptom improved. The review decided that the main cause of the injury is traffic accidents and the cineangiogram is important to find the arterial damage and the monitoring of superficial temporal arterial pressure is helpful to avoid the brain ischemia during operation. PMID- 3043168 TI - [Pulmonary flow-resistance relation in allografts ten days after single lung transplantation in dogs]. AB - In order to evaluate the applicability of single lung transplantation as a treatment of pulmonary hypertension, I investigated the relationships between the pulmonary flow and its resistance in allografts ten days after single lung transplantation in dogs. Fourteen dogs underwent single lung transplantation. All dogs received azathioprine (50 mg; given orally) and methylprednisolone sodium succinate (125 mg; given intravenously) every day after operation. Eight dogs survived up to ten days after operation. In the survived recipients and the five healthy dogs, pulmonary flow-resistance relations were investigated. Pulmonary flow was exactly regulated by the pump system, which drained from both cavae and returned to right atrium via azygos vein. The pulmonary artery of native lung or right lung was clamped prior to the measurement. Pulmonary vascular resistance was recorded with the increase of pulmonary flow at the range from 0.3 L/min. to 2.0 L/min. by 0.1 L/min. Lung water was measured by means of Wood's method. Radiographical and pathological examinations were appended. The five allografts received complete studies (successful group) were twice lung water of the healthy lungs (control group) (successful group; 95.6 +/- 16.7 g, control group; 47.8 +/- 7.5 g), and chest roentgenograms of successful group showed mild or moderate consolidation. The other allo-grafts (unsuccessful group), including ones died before investigations, were four times lung water of control group (unsuccessful group; 211.0 +/- 89.6 g), and the chest roentgenograms showed severe consolidation. But any difference was not found in any pulmonary flow between the pulmonary resistance in successful group and in control group. These results show that the allografts, which is under about twice lung water of normal lung, maintains enough pulmonary vascular function, and I concluded that single lung transplantation can be an effective treatment for pulmonary hypertension. PMID- 3043171 TI - [Twenty-four-hour liver preservation using artificial blood substitutes]. PMID- 3043172 TI - Tripartite streptokinase gene fusion vectors for gram-positive and gram-negative procaryotes. AB - A specific 1,596 bp HincII fragment ('skc) from the chromosome of Streptococcus equisimilis contains an active streptokinase (SK) gene (skc) lacking, in addition to the expression signals, codons 1 through 39 of wild-type skc but retaining the remainder of the skc coding sequence together with the transcription terminator. Using this fragment as an indicator gene, we constructed two types of vectors which in appropriate hosts resulted in the synthesis of SK fusion proteins after insertional activation of 'skc. The first type are open reading frame (ORF) vectors in which 'skc was inserted into pUC18 out of frame with respect to lacZ', thus conferring an SK-negative phenotype. Any DNA fragments representing ORFs inserted between the lacZ' expression signals and 'skc such that the skc reading frame was restored resulted in the production of tripartite proteins which exhibited SK activity. The second type of vector, which functioned in both gram positive and gram-negative bacteria, used the streptococcal speA expression and secretion signals in front of the ORF to activate 'skc insertionally. Using a large fragment from the chymosin gene as the target sequence, the usefulness of these vectors for studying foreign gene expression in streptococci as well as Escherichia coli was demonstrated. PMID- 3043174 TI - Effect of DNA sequence changes on UV mutability of a purine anticodon triplet of glyU cloned on M13 phage. AB - Mutant forms of the glyU (glycyl tRNA) gene cloned in M13mp8 were subjected to uninduced targeted UV mutagenesis; i.e. phage particles were irradiated and used to infect unirradiated umuC+ or irradiated umuC mutant cells. The irradiated phage carried GAG at the anticodon triplet and transitions to GAA were scored. The uninduced targeted mutation rate was reduced by altering the sequence of the gene in the vicinity of the target purine (Pu) residue. In particular a triplet of pyrimidines (PyPyPy) 5' to the target G was changed to PyPuPy in order to prevent formation of cyclcobutane and 6-4 pyrimidine dimers close to the target. On this basis we suggest a mechanism for one type of uninduced regionally targeted UV mutagenesis. PMID- 3043173 TI - An asparaginase of Aspergillus nidulans is subject to oxygen repression in addition to nitrogen metabolite repression. AB - Of five amidohydrolase activities subject to nitrogen metabolite repression in Aspergillus nidulans, L-asparaginase shows clearest evidence of also being subject to repression by atmospheric oxygen. Such oxygen repressibility is only evident under nitrogen metabolite derepressed conditions. Asparaginase levels are also considerably elevated by areA300, an altered function allele of the positive acting wide domain regulatory gene areA mediating nitrogen metabolite repression and are drastically reduced by loss of function mutations in areA. A. nidulans has two L-asparaginase enzymes and it has been shown by the use of appropriate mutants that these regulatory effects are exerted on the expression of that specified by the ahrA gene but probably not that specified by the apnA gene. PMID- 3043175 TI - Isolation and characterization of a variant myoblast cell line that is temperature sensitive for differentiation. AB - A new variant rat myogenic cell line, ts485, was isolated by subcloning the cell line ts3b2 (H. T. Nguyen, R. M. Medford, and B. Nadal-Ginard, Cell 34:281-293, 1983). Unlike the progenitor cell line, ts485 was thermosensitive for differentiation. Experiments with conditioned medium suggested that diffusible extracellular factors were not involved in dictating the differential phenotypes of ts485 cells cultured at the permissive and nonpermissive temperatures. Temperature shift experiments performed on cultures of ts485 cells indicated that the temperature-sensitive lesion was in a factor active during the growth phase and required to trigger a cascade of events leading to terminal differentiation. PMID- 3043176 TI - RNA11 protein is associated with the yeast spliceosome and is localized in the periphery of the cell nucleus. AB - The yeast rna mutations (rna2 through rna10/11) are a set of temperature sensitive mutations that result in the accumulation of pre-mRNAs at the nonpermissive temperature. Most of the yeast RNA gene products are involved in and essential for mRNA splicing in vitro, suggesting that they code for components of the splicing machinery. We tested this proposal by using an in vitro-synthesized RNA11 protein to complement the temperature-sensitive defect of the rna11 extract. During the in vitro complementation, the input RNA11 protein was associated with the 40S spliceosome and a 30S complex, suggesting that the RNA11 protein is indeed a component of the spliceosome. The formation of the RNA11-associated 30S complex did not require any exogenous RNA substrate, suggesting that this 30S particle is likely to be a preassembled complex involved in splicing. The RNA11-specific antibody inhibited the mRNA splicing in vitro, confirming the essential role of the RNA11 protein in mRNA splicing. Finally, using the anti-RNA11 antibody, we localized the RNA11 protein to the periphery of the yeast nucleus. PMID- 3043177 TI - Effect of limited homology on gene conversion in a Saccharomyces cerevisiae plasmid recombination system. AB - Plasmids containing heteroallelic copies of the Saccharomyces cerevisiae HIS3 gene undergo intramolecular gene conversion in mitotically dividing S. cerevisiae cells. We have used this plasmid system to determine the minimum amount of homology required for gene conversion, to examine how conversion tract lengths are affected by limited homology, and to analyze the role of flanking DNA sequences on the pattern of exchange. Plasmids with homologous sequences greater than 2 kilobases have mitotic exchange rates as high as 2 x 10(-3) events per cell per generation. As the homology is reduced, the exchange rate decreases dramatically. A plasmid with 26 base pairs (bp) of homology undergoes gene conversion at a rate of approximately 1 x 10(-10) events per cell per generation. These studies have also shown that an 8-bp insertion mutation 13 bp from a border between homologous and nonhomologous sequences undergoes conversion, but that a similar 8-bp insertion 5 bp from a border does not. Examination of independent conversion events which occurred in plasmids with heteroallelic copies of the HIS3 gene shows that markers within 280 bp of a border between homologous and nonhomologous sequences undergo conversion less frequently than the same markers within a more extensive homologous sequence. Thus, proximity to a border between homologous and nonhomologous sequences shortens the conversion tract length. PMID- 3043179 TI - Transcriptional modulation of transin gene expression by epidermal growth factor and transforming growth factor beta. AB - Transin is a transformation-associated gene which is expressed constitutively in rat fibroblasts transformed by a variety of oncogenes and in malignant mouse skin carcinomas but not benign papillomas or normal skin. It has been demonstrated that, in nontransformed Rat-1 cells, transin RNA expression is modulated positively by epidermal growth factor (EGF) and negatively by transforming growth factor beta (TGF-beta); other peptide growth factors were found to have no effect on transin expression. Results presented here indicate that both protein synthesis and continuous occupancy of the EGF receptor by EGF were required for sustained induction of transin RNA. Treatment with TGF-beta inhibited the ability of EGF to induce transin, whether assayed at the transcriptional level by nuclear run-on analysis or at the level of transin RNA accumulation by Northern (RNA) blot analysis of cellular RNA. TGF-beta both blocked initial induction of transin transcription by EGF and halted established production of transin transcripts during prolonged treatment. These results suggest that TGF-beta acts at the transcriptional level to antagonize EGF-mediated induction of transin gene expression. PMID- 3043178 TI - Relationship among guanine nucleotide exchange, GTP hydrolysis, and transforming potential of mutated ras proteins. AB - The effect of a series of mutations on the transforming potential of normal human rasH has been compared with their effects on GTPase and guanine nucleotide exchange rates of p21. The mutation Val-146 resulted in partial activation of transforming potential which could be attributed to a greater than 1,000-fold increased rate of nucleotide exchange in the absence of an effect on GTPase. In contrast, the more modest enhancement of exchange rate (approximately 100-fold) which resulted from the mutation Met-14 did not affect biological activity. The partially activating mutation Thr-59 was found to result in both a 5-fold reduction in GTPase and a 10-fold increase in nucleotide exchange. However, the nontransforming mutant Ile-59 displayed a comparable decrease in GTPase without an effect on nucleotide exchange. The activating effect of the Thr-59 mutation may thus represent a combined effect of reduced GTPase and increased exchange. Similarly, the strongly activating mutation Leu-61 resulted in a fivefold increase in nucleotide exchange in addition to decreased GTPase, whereas weakly activating mutations at position 61 (Trp and Pro) resulted only in decreased GTPase without affecting nucleotide exchange rates. Finally, combining the two mutations Met-14 and Ile-59, which alone had no effect on biological activity, yielded a double mutant with a 20-fold increased transforming potential, demonstrating a synergistic effect of these two mutations. Overall, these results indicate that large increases in nucleotide exchange can activate ras transforming potential in the absence of decreased GTPase and that relatively modest increases in nucleotide exchange can act synergistically with decreased GTPase to contribute to ras activation. PMID- 3043180 TI - c-myc and c-myb protein degradation: effect of metabolic inhibitors and heat shock. AB - The proteins encoded by both viral and cellular forms of the c-myc oncogene have been previously demonstrated to have exceptionally short in vivo half-lives. In this paper we report a comparative study on the parameters affecting turnover of nuclear oncoproteins c-myc, c-myb, and the rapidly metabolized cytoplasmic enzyme ornithine decarboxylase. The degradation of all three proteins required metabolic energy, did not result in production of cleavage intermediates, and did not involve lysosomes or ubiquitin. A five- to eightfold increase in the half-life of c-myc proteins, and a twofold increase in the half-life of c-myb proteins was detected after heat-shock treatment at 46 degrees C. In contrast, heat shock had no effect on the turnover of ornithine decarboxylase. Heat shock also had the effect of increasing the rate of c-myc protein synthesis twofold, whereas c-myb protein synthesis was decreased nearly fourfold. The increased stability and synthesis of c-myc proteins led to an overall increase in the total level of c myc proteins in response to heat-shock treatment. Furthermore, treatments which reduced c-myc and c-myb protein turnover, such as heat shock and exposure to inhibitors of metabolic energy production, resulted in reduced detergent solubility of both proteins. The recovery from heat shock, as measured by increased turnover and solubility, was energy dependent and considerably more rapid in thermotolerant cells. PMID- 3043181 TI - Mutational analysis of centromere DNA from chromosome VI of Saccharomyces cerevisiae. AB - Saccharomyces cerevisiae centromeres have a characteristic 120-base-pair region consisting of three distinct centromere DNA sequence elements (CDEI, CDEII, and CDEIII). We have generated a series of 26 CEN mutations in vitro (including 22 point mutations, 3 insertions, and 1 deletion) and tested their effects on mitotic chromosome segregation by using a new vector system. The yeast transformation vector pYCF5 was constructed to introduce wild-type and mutant CEN DNAs onto large, linear chromosome fragments which are mitotically stable and nonessential. Six point mutations in CDEI show increased rates of chromosome loss events per cell division of 2- to 10-fold. Twenty mutations in CDEIII exhibit chromosome loss rates that vary from wild type (10(-4)) to nonfunctional (greater than 10(-1)). These results directly identify nucleotides within CDEI and CDEIII that are required for the specification of a functional centromere and show that the degree of conservation of an individual base does not necessarily reflect its importance in mitotic CEN function. PMID- 3043182 TI - Identification of a Ty1 regulatory sequence responsive to STE7 and STE12. AB - Ty1 activation of gene expression observed in haploid cell types of Saccharomyces cerevisiae requires the STE7 and STE12 gene products. An activator sequence within Ty1 that is responsive to these two regulators has been defined. Complex formation between a factor in whole-cell extracts and the DNA regulatory element showed the same dependence on the STE7 and STE12 gene products as did reporter gene expression. Base pair substitutions within the binding site abolished the ability to form the factor-DNA complex and to activate gene expression. The correlation between complex formation and reporter gene expression indicates that factor binding to the cis-acting element is essential for gene activation. Because the predicted protein for the STE7 gene product is homologous to protein kinases, we suggest that protein phosphorylation may directly or indirectly regulate formation of this DNA-protein complex. PMID- 3043183 TI - Autocatalytic activities of intron 5 of the cob gene of yeast mitochondria. AB - The terminal intron of the mitochondrial cob gene of Saccharomyces cerevisiae can undergo autocatalytic splicing in vitro. Efficient splicing of this intron required a high concentration of monovalent ion (1 M). We found that at a high salt concentration this intron was very active and performed many of the reactions described for other group I introns. The rate of the splicing reaction was dependent on the choice of the monovalent ion; the reaction intermediate, the intron-3' exon molecule, accumulated in NH4Cl but not in KCl. In addition, the intron was more reactive in KCl, accumulating in two different circular forms: one cyclized at the 5' intron boundary and the other at 236 nucleotides from the 5' end. These circular forms were able to undergo the opening and recyclization reactions previously described for the Tetrahymena rRNA intron. Cleavage of the 5' exon-intron boundary by the addition of GTP did not require the 3' terminus of the intron and the downstream exon. An anomalous guanosine addition at the 3' exon and at the middle of the intron was also detected. Hence, this intron, which requires a functional protein to splice in vivo, demonstrated a full spectrum of characteristic reactions in the absence of proteins. PMID- 3043184 TI - Identification of the sites of action for regulatory genes controlling the amdS gene of Aspergillus nidulans. AB - The amdS gene of Aspergillus nidulans, which encodes an acetamidase enzyme, is positively regulated by the trans-acting genes amdR, facB, amdA, and areA. Sequence changes in several cis-acting mutations in the 5' region of the gene which specifically affect amdS regulation were determined. The amdI9 mutation, which results in increased facB-dependent acetate induction, is due to a single base change at base pair -210 relative to the start point of translation. The amdI93 mutation, which abolishes amdR-dependent omega-amino acid induction, is a deletion of base pairs -181 to -151. The amdI66 mutation, which causes increased gene activation in strains carrying amdA regulatory gene mutations, is a duplication of base pairs -107 to -90. Transformation of A. nidulans can generate transformants containing multiple integrated copies of plasmid sequences. When these plasmids carry a potential binding site for a regulatory gene product, growth on substrates whose catabolism requires genes activated by that regulatory gene can be reduced, apparently because of titration of the regulatory gene product. Introduction of 5' amdS sequences via cotransformation into strains of various genotypes was used to localize sequences apparently involved in binding of the products of the amdR, amdA, and facB genes. The position of these sequences is in agreement with the positions of the specific cis-acting mutations. Consistent with these results, a transformant of A. nidulans derived from a plasmid deleted for sequences upstream from -111 was found to have lost amdR- and facB-mediated control but was still regulated by the amdA gene. In addition, amdS expression in this transformant was still dependent on the areA gene. PMID- 3043185 TI - Systematic binding analysis of the insulin gene transcription control region: insulin and immunoglobulin enhancers utilize similar transactivators. AB - The 5' regulatory region (-345 to +1) of the rat insulin I gene (Ins-I) was examined for binding to cellular factors with short oligodeoxynucleotide probes. Over 40 binding species were detected. The binding profiles were specific for each cell type studied. We characterized the factors binding two elements crucial for enhancer activity (the Nir and Far boxes) which bear sequence similarity to the microE1, microE2, and microE3 elements of the immunoglobulin heavy-chain enhancer. The Nir box binds three cellular factors that display preferential affinities for microE1, microE2, or microE3, and the Far box binds two factors related to microE2 or microE3. The insulin gene enhancer was mutated at the Nir box element to reflect the sequences of microE1, microE2, or microE3. Ins-microE2 was fully active, Ins-microE3 was partially active, and Ins-microE1 was inactive. Thus, factors similar or identical to nuclear factor NF-microE1, NF-microE2, or NF-microE3 may play a role in the activity of the insulin gene enhancer. PMID- 3043187 TI - Molecular tools for inactivating a yeast enzyme in vivo. AB - As part of an effort to develop a new means of inducibly inactivating cellular proteins in vivo, three monoclonal antibodies which neutralize yeast alcohol dehydrogenase (ADH) activity were isolated and characterized with respect to criteria important for the inactivation strategy. The significance of these criteria is considered, and a general means of generating appropriate antibodies is suggested. All three antibodies described here were specific for ADH I; they did not recognize the closely related isozyme ADH II in a plate-binding assay and did not immunoprecipitate molecules other than ADH from a Saccharomyces cerevisiae extract. Neutralization occurred in a yeast extract and, for two antibodies, was blocked by high concentrations of the coenzyme NAD+. This finding suggests that the antibodies may block enzyme activity by stabilizing an inactive form of ADH lacking bound NAD+. These results provide a foundation for the use of these antibodies to inactivate ADH in vivo. PMID- 3043186 TI - Sp1, a CAAT-binding factor, and the adenovirus major late promoter transcription factor interact with functional regions of the gamma-fibrinogen promoter. AB - To study the factors which influence the coordinately and developmentally regulated expression of the three adjacent fibrinogen genes, we have defined the functional regions of the gamma-fibrinogen promoter and the proteins which bind to them. Using a series of 5' and internal deletion mutations, we found that sequences between 88 and 43 base pairs (bp) upstream of the gamma-fibrinogen transcription initiation site functioned in cis to direct properly initiated mRNA accumulation in transfected hepatocytes. The efficient function of these sequences was highly distance dependent, since transcriptional activity decreased by 92% when they were moved 32 bp upstream of the TATA box. We demonstrated that two known and one putative transcriptional factors interacted with this 47-bp sequence. The transcription factor Sp1 interacted with sequences between -51 and 46 as demonstrated by protection from DNase I digestion with the purified protein. Directly adjacent to the Sp1 site, between nucleotides -66 and -53, there was a sequence which bound a CAAT-binding factor. Finally, sequences just 5' to the CAAT factor-binding site interacted with the adenovirus major late transcriptional factor as previously demonstrated. Internal deletion mutations which disrupt these interactions diminished the activity of the promoter in vivo. One consequence of the interaction of these proteins is that a bend is placed in the DNA at or near their sites of interaction. PMID- 3043189 TI - Third form of the precursor to the major merozoite surface antigens of Plasmodium falciparum. AB - The precursor to the major merozoite surface antigens of Plasmodium falciparum appears to be encoded by two distinctly different (dimorphic) alleles able to undergo limited recombination. We analyzed 18 previously uncharacterized P. falciparum isolates to test the dimorphic model. All but one, a Thailand isolate, conformed to the dimorphic model, and this isolate conformed to the dimorphic model in all but variable block 2. Sequence analysis revealed that block 2 of isolate CSL2 was a third form. Hence, the dimorphic model is not strictly correct. Recombination between alleles was found only within two conserved blocks near the 5' end of the gene. PMID- 3043188 TI - B-raf, a new member of the raf family, is activated by DNA rearrangement. AB - Complementary DNA clones of a putative transforming gene were isolated from NIH 3T3 cells transformed with human Ewing sarcoma DNA. The gene was termed B-raf because it is related to but distinct from c-raf and A-raf. It appears that substitution in the amino-terminal portion of the normal B-raf protein confers transforming activity to the gene. PMID- 3043191 TI - Molecular cloning of the cDNA for a mutant mouse ribonucleotide reductase M1 that produces a dominant mutator phenotype in mammalian cells. AB - Mammalian ribonucleotide reductase is regulated by the binding of dATP and other nucleotide effectors to allosteric sites on subunit M1. Using mRNA from a mutant mouse T-lymphoma (S49) cell line, we have isolated a cDNA which encodes an altered, dATP feedback-resistant subunit M1. The mutant cDNA contains a single point mutation (a G-to-A transition) at codon 57, converting aspartic acid to asparagine. Proof that this mutation is responsible for the phenotype of dATP feedback resistance is provided by the following evidence. (i) The mutation was detected only in mutant S49 cells containing dATP feedback-resistant ribonucleotide reductase and not in wild-type or other mutant S49 cells. (ii) Transfection of Chinese hamster ovary cells with an expression plasmid containing the mutant M1 cDNA resulted in the production of dATP feedback-resistant ribonucleotide reductase. Transfected CHO cells expressing the mutant M1 cDNA exhibited a 15- to 25-fold increase in the frequency of spontaneous mutation to 6 thioguanine resistance, confirming that dATP feedback-resistant ribonucleotide reductase produces a mutator phenotype in mammalian cells. The availability of a cDNA which encodes dATP feedback-resistant subunit M1 thus provides a means of manipulating by transfection the frequency of spontaneous mutation in mammalian cells. PMID- 3043190 TI - LEU3 of Saccharomyces cerevisiae activates multiple genes for branched-chain amino acid biosynthesis by binding to a common decanucleotide core sequence. AB - LEU3 of Saccharomyces cerevisiae encodes an 886-amino-acid polypeptide that regulates transcription of a group of genes involved in leucine biosynthesis and has been shown to bind specifically to a 114-base-pair DNA fragment of the LEU2 upstream region (P. Friden and P. Schimmel, Mol. Cell. Biol. 7:2707-2717, 1987). We show here that, in addition to LEU2, LEU3 binds in vitro to sequences in the promoter regions of LEU1, LEU4, ILV2, and, by inference, ILV5. The largely conserved decanucleotide core sequence shared by the binding sites in these genes is CCGGNNCCGG. Methylation interference footprinting experiments show that LEU3 makes symmetrical contacts with the conserved bases that lie in the major groove. Synthetic oligonucleotides (19 to 29 base pairs) which contain the core decanucleotide and flanking sequences of LEU1, LEU2, LEU4, and ILV2 have individually been placed upstream of a LEU3-insensitive test promoter. The expression of each construction is activated by LEU3, although the degree of activation varies considerably according to the specific oligonucleotide which is introduced. A promoter construction with substitutions in the core sequence remains LEU3 insensitive, however. One of the oligonucleotides (based on a LEU2 sequence) was also tested and shown to confer leucine-sensitive expression on the test promoter. The results demonstrate that only a short sequence element is necessary for LEU3-dependent promoter binding and activation and provide direct evidence for an expanded repertoire of genes that are activated by LEU3. PMID- 3043192 TI - Effect of basic and nonbasic amino acid substitutions on transport induced by simian virus 40 T-antigen synthetic peptide nuclear transport signals. AB - A previous study demonstrated the ability of a synthetic peptide homologous to the simian virus 40 T-antigen nuclear transport signal to induce the nuclear transport of carrier proteins and the dependence of peptide-induced transport on a positive charge at the lysine corresponding to amino acid 128 of T antigen. In this investigation synthetic peptides were utilized to examine the effect on transport of amino acid substitutions within the T-antigen nuclear transport signal. Nuclear transport was evaluated by immunofluorescence after microinjection of protein-peptide conjugates into the cytoplasm of mammalian cells. Substitution of other basic amino acids at position 128 revealed a hierarchy for nuclear transport. The rate of nuclear transport was most rapid when a lysine was at position 128 followed in descending order by arginine, D lysine, ornithine, and p-aminophenylalanine. Peptide-induced nuclear transport was dependent upon a positively charged amino acid at positions 128 and 129, since substitutions of neutral asparagines at these positions abolished transport. However, partial transport was observed with the peptide having an asparagine at position 128 when a high number of peptides were conjugated to the carrier protein. PMID- 3043193 TI - Diversity among beta-tubulins: a carboxy-terminal domain of yeast beta-tubulin is not essential in vivo. AB - Sequences of genes for beta-tubulins from many different organisms demonstrate that they encode highly conserved proteins but that these proteins diverge considerably at their carboxyl termini. The patterns of interspecies conservation of this diversity suggest that it may have functional significance. We have taken advantage of the properties of Saccharomyces cerevisiae to test this hypothesis in vivo. The sole beta-tubulin gene of this species is one of the most divergent of all beta-tubulins and encodes 12 amino acids which extend past the end of most other beta-tubulin molecules. We have constructed strains in which the only beta tubulin gene is an allele lacking these 12 codons. We show here that this carboxy terminal extension is not essential. The absence of these 12 amino acids had no effect on a number of microtubule-dependent functions, such as mitotic and meiotic division and mating. It did confer dominant supersensitivity to a microtubule-depolymerizing drug. PMID- 3043194 TI - ACE1 regulates expression of the Saccharomyces cerevisiae metallothionein gene. AB - Copper resistance in Saccharomyces cerevisiae is mediated, in large part, by the CUP1 locus, which encodes a low-molecular-weight, cysteine-rich metal-binding protein. Expression of the CUP1 gene is regulated at the level of transcriptional induction in response to high environmental copper levels. This report describes the isolation of a yeast mutant, ace1-1, which is defective in the activation of CUP1 expression upon exposure to exogenous copper. The ace1-1 mutation is recessive and lies in a genetic element that encodes a trans-acting CUP1 regulatory factor. The wild-type ACE1 gene was isolated by in vivo complementation and restores copper inducibility of CUP1 expression and copper resistance to the otherwise copper-sensitive ace1-1 mutant. Linkage analysis and gene deletion experiments verified that this gene represents the authentic ACE1 locus. ACE1 maps to the left arm of chromosome VII, 9 centimorgans centromere distal to lys5. The ACE1 gene appears to play a direct or indirect positive role in activation of CUP1 expression in response to elevated copper concentrations. PMID- 3043195 TI - Bent DNA functions as a replication enhancer in Saccharomyces cerevisiae. AB - Previous studies have demonstrated that bent DNA is a conserved property of Saccharomyces cerevisiae autonomously replicating sequences (ARSs). Here we showed that bending elements are contained within ARS subdomains identified by others as replication enhancers. To provide a direct test for the function of this unusual structure, we analyzed the ARS activity of plasmids that contained synthetic bent DNA substituted for the natural bending element in yeast ARS1. The results demonstrated that deletion of the natural bending locus impaired ARS activity which was restored to a near wild-type level with synthetic bent DNA. Since the only obvious common features of the natural and synthetic bending elements are the sequence patterns that give rise to DNA bending, the results suggest that the bent structure per se is crucial for ARS function. PMID- 3043197 TI - TRK1 encodes a plasma membrane protein required for high-affinity potassium transport in Saccharomyces cerevisiae. AB - We identified a 180-kilodalton plasma membrane protein in Saccharomyces cerevisiae required for high-affinity transport (uptake) of potassium. The gene that encodes this putative potassium transporter (TRK1) was cloned by its ability to relieve the potassium transport defect in trk1 cells. TRK1 encodes a protein 1,235 amino acids long that contains 12 potential membrane-spanning domains. Our results demonstrate the physical and functional independence of the yeast potassium and proton transport systems. TRK1 is nonessential in S. cerevisiae and maps to a locus unlinked to PMA1, the gene that encodes the plasma membrane ATPase. Haploid cells that contain a null allele of TRK1 (trk1 delta) rely on a low-affinity transporter for potassium uptake and, under certain conditions, exhibit energy-dependent loss of potassium, directly exposing the activity of a transporter responsible for the efflux of this ion. PMID- 3043196 TI - Recessive genetic deregulation abrogates c-myc suppression by interferon and is implicated in oncogenesis. AB - In a previous study we demonstrated that many hematopoietic tumor cells are resistant to the inhibitory effects that interferon exerts on c-myc mRNA expression without losing other receptor-mediated intracellular responses (M. Einat, D. Resnitzky, and A. Kimchi, Nature [London] 313:597-600). We report here that this partial resistance was overridden in two independent stable somatic cell hybrids prepared by fusion between sensitive and resistant cells. The c-myc mRNA transcribed from the active allele of the resistant parent cell was reduced by interferon within the context of the cell hybrid. It was therefore concluded that changes in the cis-acting sequences of c-myc were not involved in this type of relaxed regulation and that resistance resulted rather from inactivation or loss of postreceptor elements which operate in trans. The growth-stimulating effect that this genetic deregulation might have on cells was tested in experimental systems of cell differentiation in which an autocrine interferon is produced. For that purpose we isolated variant clones of M1 myeloid cells which were partially resistant to alpha and beta interferons and tested their growth behavior during in vitro-induced differentiation. The resistant clones displayed higher proliferative activity on days 2 and 3 of differentiation than did the sensitive clones, which stopped proliferating. The loss of c-myc responses to the self-produced interferon disrupted the normal cessation of growth during differentiation and therefore might lead cells along the pathway of neoplasia. PMID- 3043198 TI - cis-acting translational effects of the 5' noncoding region of c-myc mRNA. AB - We have previously shown that the 5' noncoding region of mouse c-myc mRNA has a negative effect on translational efficiency in a rabbit reticulocyte lysate (A. Darveau, J. Pelletier, and N. Sonenberg, Proc. Natl. Acad. Sci. USA 82:2315-2319, 1985). We wanted to localize and characterize the inhibitory translational element(s) in the mRNA and to study its effect in other in vitro and in vivo systems. Here we report that the restrictive element is confined to a 240 nucleotide sequence of the 5' noncoding region of mouse c-myc mRNA and that this sequence acts in cis to inhibit the translation of a heterologous mRNA. In addition, we report that the cis-inhibitory effect is also exhibited in microinjected Xenopus oocytes and wheat-germ extracts but not in HeLa cell extracts. Transfection of corresponding plasmid DNA constructs into several established cell lines did not produce the cis-inhibitory effect. A model to explain these results is presented. PMID- 3043199 TI - Processing, secretion, and biological properties of a novel growth factor of the fibroblast growth factor family with oncogenic potential. AB - We recently reported that the protein encoded in a novel human oncogene isolated from Kaposi sarcoma DNA was a growth factor with significant homology to basic and acidic fibroblast growth factors (FGFs). To study the properties of this growth factor (referred to as K-FGF) and the mechanism by which the K-fgf oncogene transforms cells, we have studied the production and processing of K-FGF in COS-1 cells transfected with a plasmid encoding the K-fgf cDNA. The results show that, unlike basic and acidic FGFs, the K-FGF protein is cleaved after a signal peptide, glycosylated, and efficiently secreted as a mature protein of 176 or 175 amino acids. Inhibition of glycosylation impaired secretion, and the stability of the secreted K-FGF was greatly enhanced by the presence of heparin in the cultured medium. We have used the conditioned medium from transfected COS 1 cells to test K-FGF biological activity. Similar to basic FGF, the K-FGF protein was mitogenic for fibroblasts and endothelial cells and induced the growth of NIH 3T3 mouse cells in serum-free medium. Accordingly, K-fgf transformed NIH 3T3 cells grew in serum-free medium, consistent with an autocrine mechanism of growth. We have also expressed the protein encoded in the K-fgf protooncogene in COS-1 cells, and it was indistinguishable in its molecular weight, glycosylation, secretion, and biological activity from K-FGF. Taken together, these results suggest that the mechanism of activation of this oncogene is due to overexpression rather than to mutations in the coding sequences. PMID- 3043200 TI - Genetic selection for mutations that reduce or abolish ribosomal recognition of the HIS4 translational initiator region. AB - A unique genetic selection was devised at the HIS4 locus to address the mechanism of translation initiation in Saccharomyces cerevisiae and to probe sequence requirements at the normal translational initiator region that might participate in ribosomal recognition of the AUG start codon. The first AUG codon at the 5' end of the HIS4 message serves as the start site for translation, and the -3 and +4 nucleotide positions flanking this AUG (AXXAUGG) correspond to a eucaryotic consensus start region. Despite this similarity, direct selection for mutations that reduce or abolish ribosomal recognition of this region does not provide any insight into the functional nature of flanking nucleotides. The only mutations identified that affected recognition of this region were alterations in the AUG start codon. Among 150 spontaneous isolates, 26 were shown to contain mutations in the AUG start codon, including all +1 changes (CUG, GUG, and UUG), all +3 changes (AUA, AUC, and AUU), and one +2 change (ACG). These seven mutations of the AUG start codon, as well as AAG and AGG constructed in vitro, were assayed for their ability to support HIS4 expression. No codon other than AUG is physiologically relevant to translation initiation at HIS4 as determined by growth tests and quantitated in his4-lacZ fusion strains. These data and analysis of other his4 alleles are consistent with a mechanism of initiation at HIS4 as proposed in the scanning model whereby the first AUG codon nearest the 5' end of the message serves as the start site for translation and points to the AUG codon in S. cerevisiae as an important component for ribosomal recognition of the initiator region. PMID- 3043201 TI - Mutational analysis of the HIS4 translational initiator region in Saccharomyces cerevisiae. AB - We have mutated various features of the 5' noncoding region of the HIS4 mRNA in light of established Saccharomyces cerevisiae and mammalian consensus translational initiator regions. Our analysis indicates that insertion mutations that introduce G + C-rich sequences in the leader, particularly those that result in stable stem-loop structures in the 5' noncoding region of the HIS4 message, severely affect translation initiation. Mutations that alter the length of the HIS4 leader from 115 to 39 nucleotides had no effect on expression, and sequence context changes both 5' and 3' to the HIS4 AUG start codon resulted in no more than a twofold decrease of expression. Changing the normal context at HIS4 5' AAUAAUGG-3' to the optimal sequence context proposed for mammalian initiator regions 5'-CACCAUGG-3' did not result in stimulation of HIS4 expression. These studies, in conjunction with comparative and genetic studies in S. cerevisiae, support a general mechanism of initiation of protein synthesis as proposed by the ribosomal scanning model. PMID- 3043202 TI - Invariant phosphorylation of the Saccharomyces cerevisiae Cdc28 protein kinase. AB - The phosphorylation level of the Saccharomyces cerevisiae Cdc28 protein remained invariant under conditions that resulted in cell cycle arrest in the G1 phase and loss of Cdc28-specific protein kinase activity when the activity was assayed in vitro. These results are in contrast to the proposed regulation of the homologous Cdc2 protein kinase of Schizosaccharomyces pombe. PMID- 3043204 TI - [Alternative nutrition of children. Its advantages and risks]. AB - Food faddism is a growing scenery. Since children are also involved in these unusual food habits of their parents, the pediatrician is faced with new nutritional problems. The consequence may be failure to thrive in infancy and childhood noticed mainly in families with strictly vegetarian food habits. Moreover the pediatrician should know the possible sequelae of all the other forms of food faddism. Only a careful nutritional history paralleling the usual medical history may then uncover the origin of a chronic failure to thrive. First and foremost infants after weaning are at special risk in respect to protein, calcium and vitamin deficiencies. PMID- 3043203 TI - A new RAS mutation that suppresses the CDC25 gene requirement for growth of Saccharomyces cerevisiae. AB - In the yeast Saccharomyces cerevisiae, the activation of adenylate cyclase requires the products of the RAS genes and of CDC25. We isolated several dominant extragenic suppressors of the yeast cdc25 mutation. They did not suppress a thermosensitive allele of the adenylate cyclase gene (CDC35). One of these suppressors was a mutated RAS2 gene in which the transition C/G----T/A at position 455 resulted in replacement of threonine 152 by isoleucine in the protein. The same mutation in a v-Ha-ras gene reduces the affinity of p21 for guanine nucleotides (L.A. Feig, B. Pan, T.M. Roberts, and G.M. Cooper, Proc. Natl. Acad. Sci. USA 83:4607-4611, 1986). These results support a model in which the CDC25 gene product is the GDP-GTP exchange factor regulating the activity of the RAS gene product. PMID- 3043205 TI - [Breast feeding as "alternative" nutrition? An orientation aid in the field spanning ideology and current status]. AB - Recommending breast feeding supports a trend "back to nature". However, there are believers in the unbiased goodness of nature, who claim effects of mothers milk, e.g. protection from the evils of civilization, that are not confirmed by current scientific knowledge. As nature is not uniquely good (it feeds the donkey and the lion), and as biology is not perfect, breast feeding does not guarantee an uneventful infancy and childhood. There is even a pathology of breast feeding that the pediatrician should be familiar with.- The proven benefits, claimed effects, and some risks of breast feeding are reviewed on the basis of research results, and conclusions are drawn to make breast feeding as uneventful an experience as possible. Potential fields of research are being identified. PMID- 3043206 TI - [Ultrasonic assessment of the development of the uterus in childhood]. AB - In 150 girls from the newborn period up to the 18. year the size of the uterus was measured by sonography. In all children the length, width, depth and volume of the uterus as well as its shape and other signs of development were determined. In the newborn period and in the following first months of life an estrogen stimulated uterus with the shape of a drop could be shown. The size of the uterus decreased till the end of the first year. The length of the uterus in the neonatal period was 4.0 +/- 0.5 cm, the volume was 3.6 +/- 1.9 cm3. In the following rest phase up to the age of 8 years the length of the uterus was 2.8 +/ 0.4 cm and the volume was 1.2 +/- 0.4 cm3. The shape of the uterus in this age group was tubular. One to two years before secondary pubertal signs could be shown, sonography already demonstrated the beginning of pubertal uterus growth. In the postpubertal period the uterus had the typical pear shape with a corpus/cervix-relation of 2:1. In the postpubertal period the length was 6.8 +/- 1 cm with a volume of 33 +/- 22 cm3. The knowledge of the normal developmental dates of the uterus in children is essential for the diagnosis of malformations and tumours of the pelvis, but also for the diagnosis of distorted puberty such as pubertas praecox, pubertas tarda, amenorrhea and growth retardation. PMID- 3043207 TI - [Elastase-alpha 1-proteinase inhibitor in diseases of the neonatal period]. AB - Elastase, a neutral protease stored in the azurophilic granules of neutrophils, is immediately released during the process of phagocytosis and rapidly bound and inactivated by alpha 1-proteinase inhibitor. This complex (E-alpha 1-PI) is highly stable and can be identified by ELISA-technique. In our study 95% of all infants with neonatal septicemia and/or meningitis had significantly increased plasma levels of E-alpha 1-PI at the time of diagnosis (n = 37). After initiation of therapy normalization of E-alpha 1-PI levels was observed in all neonates who recovered from infection. These data suggest that E-alpha 1-PI is a sensitive and rapidly responsive indicator of neonatal septicemia. In addition E-alpha 1-PI may be helpful in monitoring the course of the disease. However, the specificity of E alpha 1-PI is rather low: in patients with local infections and inflammatory processes such as neonatal pneumonia, enterocolitis and meconium aspiration E alpha 1-PI levels were also shown to be increased. In contrast, all patients with hyaline membrane disease had E-alpha 1-PI levels within the normal range. PMID- 3043208 TI - [Rothmund syndrome or Thomson syndrome. An analysis of the literature exemplified by a personal case]. AB - We report on a now 16 month old child, born with multiple bone defects. A characteristic poikiloderma leading to the diagnosis appeared after six months. A review of the literature is given and the clinical heterogeneity of the Rothmund Thomson-Syndrome is discussed. It is suggested, that the combination of the described malformations represents a Thomson Syndrome. PMID- 3043209 TI - [Mediastinal teratoma in a newborn infant]. AB - A newborn with mediastinal teratoma developed tachypnea and cyanosis within the first hours of life. Chest x-ray revealed a subtotal opacification of the left hemithorax, which at first was suspected to be caused by a cardiomegaly. Ultrasound examination demonstrated a large mass in the left hemithorax with multiple echo-free areas. This finding was suspicious for a mediastinal teratoma. The tumor which was originating from the mediastinum was apple-sized and could be resected totally by thoracotomy. Histology revealed a cystic teratoma covered with normal thymus tissue. PMID- 3043210 TI - [A principle of maximum topological similarity in molecular systematics]. AB - The paper deals with the problem of phylogenetic reconstruction on the basis of comparative analysis of features. Main attention is paid to comparison and classification of the biopolymer sequences. Different approaches to this task are critically reviewed. The novel principle of construction of tree-like classification schemes permitting subsequent evolutionary analysis is proposed. It concentrates on reconstruction of the tree with a topologic structure that is most close to topologic features, imprinted in the source distance matrix. Realization of this approach was made possible by development of the special formalism, enabling evaluation and comparison of topologic features of distance matrices and trees. PMID- 3043211 TI - [Effect of phospholipase on cation-induced transmembrane transport of DNA in Escherichia coli]. AB - Incubation of bacterial cells in 0.1 M CaCl2 at 0 degrees C considerably increases the amount of phospholipids susceptible to action of a specific enzyme of phospholipid metabolism phospholipase C (hydrolysis to diacylglycerides). In process of incubation in CaCl2 solutions at 0 degrees C the expressed activity of an endogenous enzyme phospholipase A has been registered in cellular samples. Binding of the enzyme by the cells under conditions unfavourable for phospholipids hydrolysis (0 degrees C) suppresses strongly and reversibly cellular ability to DNA transformation without affecting cellular survival. As calculated, the enzyme molecules cover about 10% of cellular surface while inhibiting 90% of transmembrane transfer. The obtained data are considered to be a solid argument supporting the important role of the membrane phospholipids in the mechanism of cation-induced DNA transfer into the cell. PMID- 3043212 TI - [Algorithms for constructing phylogenetic trees of maximum topological similarity]. AB - The paper concerns the practical realization of the maximum topologic similarity principle for phylogenetic reconstruction. This novel principle is described in the accompanying paper. Two algorithms that were embodied in the computer program allow one to find out the unique tree in case when source data admit the existence of such tree. In case if numerous parallel mutations make such precise realization impossible, algorithms allow one to obtain approximations to the maximum topologic similarity trees with a high computation efficiency. Examples illustrating use of these algorithms, as well as discussion of biological consistency of the novel concept are presented. PMID- 3043213 TI - [Structural organization of the genome of paramyxoviruses]. AB - The review summarizes the recent papers on the studies of primary structure of genome of a number of paramyxoviruses from the three genera of a family. The cited data demonstrate that despite the common principles of the genetic material arrangement shared by paramyxoviruses, they are variable in the genome, the primary structure of intragenic region, as well as the strategy of coding for some proteins. The data on the arrangement of the genetic material is discussed as useful as a criterion for classification of single stranded viruses with unsegmented genome. PMID- 3043214 TI - Molecular genetics in basic myology: a rapidly evolving perspective. AB - Myology has greatly benefited from the recent unification of concepts in molecular, cellular, and developmental biology. The interplay between intrinsic and extrinsic factors in determining the physiologic characteristics of individual myofibers has emerged as an important theme. Of special note is the manner in which the study of contractile protein gene structure and expression has contributed to our understanding of the development and ultimate plasticity of the contractile apparatus. As mechanistic models of normal myogenesis achieve increasing sophistication, the opportunities for understanding the pathogenesis of progressive muscle disfunction improve. In this article we review recent progress in basic myology which will be of interest to clinicians studying the heritable neuromuscular disorders. PMID- 3043215 TI - Molecular genetics in muscular dystrophy research: revolutionary progress. AB - The contribution of "reverse genetic" strategies to neuromuscular disease research is evident in the progression of breakthroughs that have recently culminated in the cloning of the Duchenne muscular dystrophy (DMD) cDNA. The resultant improvements in our understanding of the genetic basis of Becker muscular dystrophy (BMD) and DMD serve as models for similar investigation of other heritable disorders. These genetic advances have outpaced concurrent work on the molecular pathogenesis of the dystrophic process, with the curious result that inferences about the DMD protein's amino acid sequence have preceded any information about its function or intracellular localization. In recognition that this foundation sets the stage for the rapid elucidation of the disease's pathogenesis, we review the experimental basis of such advances, with reference to relevant progress in basic myology, pathology, and molecular biology. We conclude with a view towards the ultimate clinical implications of these experimental breakthroughs. PMID- 3043216 TI - Altered blood rheology in the pathogenesis of diabetic and other neuropathies. AB - Although a substantial literature confirms the abnormal flow properties of diabetic blood, only in a few papers has the vasculitis of diabetic neuropathy been considered to have a hemorheological cause. It is proposed that the pathogenesis of nerve lesions involves an interaction between the specialized nerve vascular system and focal ischemic lesions resulting from rheologically induced stasis. The proposition is extended into other conditions with abnormal blood rheology such as hypothyroidism, uremia, dysglobulinemia, polyarteritis nodosa, and lepromatous leprosy. It is concluded that the treatment of such polyneuropathies should include an agent which would improve the flow properties of the blood. PMID- 3043217 TI - EMG computerized analysis of localized fatigue in Duchenne muscular dystrophy. AB - EMG power spectra obtained during a sustained isometric contraction were analyzed in a group of 10 children affected by Duchenne muscular dystrophy (DMD) and compared with those obtained in a control group of 5 normal children. In myopathic subjects the isometric contraction caused an increase of the total power, a progressive increase of power of the lower frequencies, a decrease of that of the higher frequencies, and a shift downward of the median frequency. In normal children an increase of the total power without a significant median frequency shift was noted. The modifications observed in DMD children were explained by a decrement of the firing rate of the more damaged fast twitch motor units. This decrease was probably induced by a relative predominance of activity of the slow twitch motor units, which are less damaged by the pathological process. PMID- 3043218 TI - Kidney function in experimental systemic candidosis of mice. PMID- 3043219 TI - Clinical characteristics correlated with different fungi causing vulvovaginal mycosis. PMID- 3043220 TI - Regulation of the direct fungicidal action of miconazole against Candida albicans ATCC 26790. PMID- 3043221 TI - Humoral hypercalcemia of cancer. Identification of a novel parathyroid hormone like peptide. PMID- 3043224 TI - Information services in medical mycology. AB - The need for information services in medical mycology is discussed. The information service provided by the Review of Medical and Veterinary Mycology is described and compared with other information services. PMID- 3043223 TI - Adherence of Candida species to fibrin clots in vitro. AB - The adherence of six Candida species to fibrin clots was studied using a simple, in vitro technique. Yeast suspensions were incubated with fibrin clots and the number of adherent organisms quantified as follows: after washing, the clots were subjected to vortex mixing and the number of CFU's which subsequently grew on Sabourauds medium were counted. Adhesion was directly proportional to the concentration of Candida species in the suspension (r = 0.99 p less than 0.001). C. albicans and C. tropicalis exhibited marked adherence whereas C. krusei, C. gulliermondi and C. glabrata adhered less readily. C. parapsilosis was intermediate in its ability to adhere. PMID- 3043222 TI - Effect of Candida albicans cell wall components on the adhesion of the fungus to human and murine vaginal mucosa. AB - In this study, cell walls from Candida albicans were separated and chitin was isolated from these cell walls. A chitin soluble extract (CSE) prepared from the chitin inhibited in vitro adhesion of C. albicans to human epithelial vaginal cells (VEC), and blocked in vivo attachment to murine vaginal mucosa, thereby preventing candidal infection in these animals. These findings suggest that the CSE acts as an adhesin-like substance. Fractionation of CSE yielded two fractions: FI and FII, of which only FI exhibited inhibitory activity. Chemical analysis of CSE and its two fractions revealed that CSE contains over 70% of proteins, most of which were found in the non-active fraction. In addition, 3% of amino-sugars were found in the FI active fraction. Lipids were also detected in the unfractionated CSE and in both fractions. Experiments to further characterize the component(s) in the CSE inhibiting the attachment of C. albicans are in progress in our laboratory. PMID- 3043225 TI - Cryptococcus neoformans: a central nervous system isolate from an AIDS patient that is rhinotropic in a normal mouse model. AB - A strain of Cryptococcus neoformans that was isolated from the cerebrospinal fluid of a human diagnosed as having acquired immunodeficiency syndrome (AIDS), and that produced cutaneous lesions in experimentally infected, normal mice is described. Although no unusual cutaneous manifestations were noted in the patient's records, this isolate of C. neoformans proved to be dermotropic when injected intravenously into CD-1 mice. The LD50 at 28 days post infection ranged from 3.6-7.5 X 10(5) cells per mouse, and in vitro growth rate studies demonstrated that this isolate grew well at 35 degrees C and at 37 degrees C, but did not grow at 40 degrees C and higher. This isolate was rhinotropic producing large granulomatous lesions in the nasal tissues. Other cutaneous tissues affected were the periocular tissues, ears, feet and tail, although the granulomas were nodular in structure and less necrotic than the nasal lesions. The brain, lungs, liver, kidneys and spleen also were culture positive for C. neoformans. Histopathologically, each affected tissue examined had large densities of yeast cells and a chronic, granulomatous host response. Animals surviving the infection appeared to develop a commensal-type relationship with the infective yeast. This is the first report of an isolate of C. neoformans from an AIDS patient that has caused cutaneous manifestations in an animal model. The model described in this report may be useful for elucidating pathogenic mechanisms of cryptococcosis, particularly cutaneous manifestations of the disease. PMID- 3043226 TI - T-cell antigen receptor genes and T-cell recognition. AB - The four distinct T-cell antigen receptor polypeptides (alpha, beta, gamma, delta) form two different heterodimers (alpha:beta and gamma:delta) that are very similar to immunoglobulins in primary sequence, gene organization and modes of rearrangement. Whereas antibodies have both soluble and membrane forms that can bind to antigens alone, T-cell receptors exist only on cell surfaces and recognize antigen fragments only when they are embedded in major histocompatibility complex (MHC) molecules. Patterns of diversity in T-cell receptor genes together with structural features of immunoglobulin and MHC molecules suggest a model for how this recognition might occur. This view of T cell recognition has implications for how the receptors might be selected in the thymus and how they (and immunoglobulins) may have arisen during evolution. PMID- 3043227 TI - The helical repeat of double-stranded DNA varies as a function of catenation and supercoiling. AB - DNA in the cell is intertwined at several levels: one polynucleotide strand wraps helically around its complement and the double helix is in turn coiled in space. The higher-order intertwining most often takes the form of supercoiling of the helix axis, but can also be observed as the wrapping of one DNA duplex around another, as in catenation. We have investigated the relationship between intertwining at these three levels, the double helix, supercoiling, and catenation, using an approach that relies on comparative measurements of DNA linking numbers by gel electrophoresis. The method determines both the handedness of DNA catenanes and the change in helical repeat that accompanies catenation induced supercoiling. For multiply-linked catenated rings of 3.5 kilobase pairs (kb), we conclude that the double helix unwinds by two-thirds of a turn for every right-handed supercoil involved in linking the two circles. Altering the geometry of the catenanes by linking rings of dissimilar size changes the effect of catenation on helical and superhelical parameters. Our experiments used intact DNA rings, but we note that linear DNA molecules, by virtue of their subdivision into closed loops or domains in vivo, can intertwine in the same ways. PMID- 3043228 TI - Internal sequences that distinguish yeast from metazoan U2 snRNA are unnecessary for pre-mRNA splicing. AB - U2 small nuclear RNA is a highly conserved component of the eukaryotic cell nucleus involved in splicing messenger RNA precursors. In the yeast Saccharomyces cerevisiae, U2 RNA interacts with the intron by RNA-RNA pairing between the conserved branchpoint sequence UACUAAC and conserved nucleotides near the 5' end of U2 (ref. 4). Metazoan U2 RNA is less than 200 nucleotides in length, but yeast U2 RNA is 1,175 nucleotides long. The 5' 110 nucleotides of yeast U2 are homologous to the 5' 100 nucleotides of metazoan U2 (ref. 6), and the very 3' end of yeast U2 bears a weak structural resemblance to features near the 3' end of metazoan U2. Internal sequences of yeast U2 share primary sequence homology with metazoan U4, U5 and U6 small nuclear RNA (ref. 6), and have regions of complementarity with yeast U1 (ref. 7). We have investigated the importance of the internal U2 sequences by their deletion. Yeast cells carrying a U2 allele lacking 958 nucleotides of internal U2 sequence produce a U2 small nuclear RNA similar in size to that found in other organisms. Cells carrying only the U2 deletion grow normally, have normal levels of spliced mRNA and do not accumulate unspliced precursor mRNA. We conclude that the internal sequences of yeast U2 carry no essential function. The extra RNA may have a non-essential function in efficient ribonucleoprotein assembly or RNA stability. Variation in amount of RNA in homologous structural RNAs has precedence in ribosomal RNA and RNaseP. PMID- 3043229 TI - Increased virulence of Yersinia pseudotuberculosis by two independent mutations. AB - A chromosomally encoded protein, which mediates invasion into HeLa cells was recently identified in Yersinia pseudotuberculosis. The role of this protein (invasin) in the virulence process was not, however, investigated. We show that mutation of the invasin gene in Y. pseudotuberculosis abolishes the ability of the bacteria to invade HeLa cells. When mice were challenged by intraperitoneal injection both the mutant and the wild-type strain produced infections of similar virulence but mutant showed a slower rate of infection after oral challenge. A double mutant, carrying an additional mutation in the gene coding for the Yop1 protein, was also constructed. The double mutant was significantly more virulent than either the wild-type or the corresponding single mutants. Y. pestis, in contrast to Y. pseudotuberculosis lacks the ability to express either invasin or Yop1, sequence analysis of the yopA gene from both Y. pestis and Y. pseudotuberculosis shows that the yopA gene of Y. pestis contains a point mutation leading to a reading-frame shift. When the yopA+ gene was introduced into Y. pestis the virulence of this strain was reduced. These results may provide insight into the rise and fall of plague epidemics caused by Y. pestis. PMID- 3043232 TI - US looking for short cuts to speed drug approval. PMID- 3043230 TI - Identification of a protein encoded by the vpu gene of HIV-1. AB - Human immunodeficiency virus 1 (HIV-1) is the aetiological agent of AIDS. The virus establishes lytic, latent and non-cytopathic productive infection in cells in culture. The complexity of virus-host cell interaction is reflected in the complex organization of the viral genome. In addition to the genes that encode the virion capsid and envelope proteins and the enzymes required for proviral synthesis and integration common to all retroviruses, HIV-1 is known to encode at least four additional proteins that regulate virus replication, the tat, art, sor and 3' orf proteins, as well as a protein of unknown function from the open reading frame called R. Close examination of the nucleic acid sequences of the genomes of multiple HIV isolates raised the possibility that the virus encodes a previously undetected additional protein. Here we report that HIV-1 encodes a ninth protein and that antibodies to this protein are detected in the sera of people infected with HIV-1. This protein distinguishes HIV-1 isolates from the other human and simian immunodeficiency viruses (HIV-2 and SIV) that do not have the capacity to encode a similar protein. PMID- 3043231 TI - The function and structure of the metal coordination sites within the glucocorticoid receptor DNA binding domain. AB - The glucocorticoid receptor enhances or represses transcription by binding to specific DNA sequences termed glucocorticoid response elements, or GREs. Studies of cloned glucocorticoid receptors reveal that the protein is organized as functional domains, in an arrangement that appears to be common among members of the steroid receptor family. A segment near the centre of the gene specifies DNA binding activity in vitro and contains two sequence motifs similar to 'zinc fingers' found in Xenopus transcription factor IIIA (TFIIIA). Such sequence motifs have been identified in nucleic acid binding proteins from a wide range of organisms. Steroid receptor protein fingers are proposed to bind zinc through two pairs of conserved cysteine residues. We report here that a protein of relative molecular mass 19,000 (Mr = 19 K) encompassing the DNA-binding domain of the glucocorticoid receptor that has been overexpressed in Escherichia coli and purified to homogeneity reversibly ligates two Zn(II) or Cd(II) ions. We show that metal ions are required for specific DNA binding and proper folding. Using EXAFS (extended X-ray absorption fine structure) and visible light spectroscopies, we find that each Zn atom is coordinated in a tetrahedral arrangement by four cysteines. PMID- 3043233 TI - Tests for new AIDS treatment begin in three clinics. PMID- 3043235 TI - [Digoxin]. PMID- 3043234 TI - The endocrinology of sunlight and darkness. Complementary roles of vitamin D and pineal hormones. AB - Information from autoradiographic studies and follow-up indicates that 1,25(OH)2 vitamin D3 (soltriol) is a somatotrophic activator and modulator, regulated by the amount of sunshine and the endocrine status of the individual, with the purpose of promoting development, reproduction, and maintenance of life. Regulation of calcium homeostasis is only one of its many functions. A close link to the pineal hormone system is apparent. Evidence supports the new concept that the skin-derived hormone of sunlight and the pineal hormone(s) of darkness are messengers with comprehensive actions on endocrine, autonomic, sensory, skeletal, and motor functions. In a complementary fashion, both hormone systems appear to correlate biological activities with the daily and seasonal changes of our solar environment. PMID- 3043236 TI - [The connection between depression and dementia in the elderly]. PMID- 3043237 TI - [Pleuropulmonary disorders in rheumatoid arthritis]. PMID- 3043238 TI - Spontaneous acute lymphoblastic leukemia in Sprague-Dawley rats. I. Immunogenetic analysis of four individual leukemias. AB - The antigenic phenotyping of four individual spontaneous ALLs of SD strain revealed practically complete concordance of RT1 class I expression with PBL of SD strain. RT5 antigen as well as class II and SIg markers were not proven. T cell markers defined by MoAbs OX7 (Thy 1.1) and W3/13 (T cell common) were detected on all four ALL tested. PMID- 3043239 TI - [Syndrome of painful muscle fasciculations. Case report and differential diagnostic review]. AB - The case of a patient with spontaneous painful muscular activity is described. We diagnosed the muscular pain fasciculation syndrome. Biopsy of the sural nerve and electrophysiological examination provided evidence of the neurogenic nature of the disturbance showing a remarkable axonal degeneration. In addition, when immunohistochemical studies were performed, the detection of immunoglobulins and complement components suggested an immunological process in the etiology of the disease. The patient made a good response to carbamazepine therapy. This effect is found in various syndromes of abnormal continuous muscle activity. Clinical, neurophysiological and morphological features of this case are discussed in relation of pathogenesis, possible drug effects and related diseases. PMID- 3043240 TI - [Moyamoya syndrome]. PMID- 3043242 TI - [Polyneuropathy and myopathy in oxalosis]. PMID- 3043241 TI - [Ventricular tachycardias of the torsade de pointes type in transtentorial herniation]. PMID- 3043243 TI - Solitary intracranial extracerebral glioma. AB - The authors present the case of a 65-year-old woman with a solitary extracerebral glioma. The tumour originated from the falx in the left fronto-parietal region near the paracentral lobule. Because it was well delineated and was completely outside the brain it was thought at operation to be a meningioma or a metastasis. Histologically the tumour could be classified as an oligodendroglioma (WHO grade II). Intracranial extracerebral gliomas so far described are most frequently located in the vicinity of the sylvian fissure. Involvement of the dura has only been observed in three cases. This case is to the best of our knowledge the only one in which the tumour was situated in the longitudinal fissure having originated from the falx. Extracerebral gliomas are thought to arise from heterotropic nests of glial cells in the leptomeninges. PMID- 3043244 TI - [Treatment of atypical post-traumatic and postoperative facial neuralgias by chronic stimulation. Apropos of 2 cases, with review of the literature]. AB - This article describes the case of two patients suffering from deafferentation pain after surgery or traumatic lesions in the area of the peripheric trigeminal branches on the Gasserian ganglion. Chronic electrical stimulation by an electrode implanted in the Gasserian ganglion led to a good permanent result in both cases, within a follow-up period of one year and one year and a half, respectively. The pre-operative test was carried out percutaneously via the foramen ovale, the definitive implantation by surgery with subtemporal access. This method can only be used when at least part of the ganglion cells of the Gasserian ganglion are intact. According to identical observations by other authors, it is mostly adequate for surgical and traumatic trigeminal lesions, whereas for pain due to herpes zoster, the stimulation of specific thalamus nuclei is a much better method. PMID- 3043246 TI - [Papilloma of the choroid plexus of the lateral ventricle without generalized hydrocephalus]. AB - A benign papilloma of the choroid plexus of the left lateral ventricle is found in a five-month old infant. An important dilatation of the isolated occipital horn is observed, without generalized enlargement of the ventricles. The clinical signs are limited to a moderate macrocephaly with deviation of head and eyes to the left. The absence of global hydrocephalus is rather exceptional in such cases, where overproduction of cerebrospinal fluid, in an amount proportional to the tumoral mass, is generally admitted. Nevertheless, the literature offers no absolute proof of the role of oversecretion alone in the origin of hydrocephalus, whilst an eventual obstruction of the circulatory pathways and resorption areas of the C.S.F. must be kept in mind. The absence of hydrocephalus, in the present case, suggests that the subarachnoid spaces and arachnoid villi remained patent. PMID- 3043245 TI - [Intracerebral schwannoma. Apropos of a case]. AB - The authors report a case of intracerebral Schwannoma discovered in a 13 year old girl who had shown symptoms of increased intracranial pressure since four years and half evolving progressively to aggravation. CT scan images and operating appearance were uncharacteristic. Histological diagnosis of the tumor was only established at pathological examination. The tumor recurred 15 months after in spite of its complete extirpation. The characteristics of this extremely rare localization of Schwannoma (14 cases published in the literature) are pin-pointed by the authors. PMID- 3043247 TI - [Recurrences of supratentorial meningiomas. Apropos of 17 cases]. AB - 17 cases of recurrent meningiomas observed during a 10 years period are reviewed. Meningiomas with a second localisation different from the first were excluded from the study. The tumor recurrence occurred mostly during the first five years after the surgical procedure. One case with femoral and pulmonary metastasis is described. Several factors may play a role in the genesis of the recurrence: age, sex, the tumor localisation, CT scan, the importance of the tumor ablation, the histology. In almost all cases a radiotherapy was carried out after the second surgical procedure. This retrospective study suggests the need for multifactorial prospective analysis with clinical, paraclinical and fundamental factors. Such a study would be useful to precise the natural history of meningiomas. PMID- 3043248 TI - [Anterior sacral meningocele. Apropos of 11 cases]. AB - The authors report 11 cases of anterior sacral meningocele, cystic mass connected with lumbar sac through a sacral bony defect: 2 children and 9 adults. Clinic presentations are analysed. Examination including, X ray, ultrasonography, scanner is discussed. The authors believe that these lesions must be treated surgically because a risk of rupture in the rectum with important meningitis exists. There is also a septic risk if an error of diagnosis ends at a puncture. The transacral approach is easy and it gives the best results. PMID- 3043249 TI - [Stereotaxic guidance in open surgery of the brain]. AB - The authors describe the technique used to precisely locate, before surgical ablation, small superficial or deep seated intracerebral lesions. When they are superficial, a burrhole is placed above the lesional site, that is reconstructed by superimposing the CT image onto a plain film radiogram taken during stereotaxy. Deep seated lesions are localized by inserting a guide under stereotaxy, prior to open surgery, which will lead the neurosurgeon without error from the cortex to the lesion. 37 observations are reported where this technique lead to a greatly simplified intervention with minimal cortical damage. PMID- 3043251 TI - Improvement of IVDSA of the brachiocephalic arteries using a non-ionic contrast medium. AB - The introduction of IVDSA provides a safer method of angiographic investigation of the aortic arch and brachiocephalic arteries than conventional angiography in patients with cerebro-vascular disease. The quality of visualization however decreases. Instead of changing over to IADSA we find it worthwile to try to improve IVDSA in different ways. One of them is represented in this prospective double blind cross-over study in which we have investigated the effects of using an isotonic non-ionic contrast medium, iohexol (omnipaque), in comparison to meglumin-sodium-diatrizoate (urografin) which is widely used. Iohexol causes less severe side-effects, less severe artifacts and a better image quality. A statistical relation between parameters could not be established. A lag-effect exists for side-effects. The necessity of using low osmolar contrast media is discussed. PMID- 3043253 TI - Invariant structure in locomotion. AB - Biological systems are hypothesized to control behavior with reference to invariants, because this would allow the variable but robust accomplishment of tasks observed in biological behaviors. Invariants for legged locomotion are specified. Combined with observed properties of locomotion, they lead to predictions of forms of control for legged locomotion. PMID- 3043252 TI - Towards an advisor for MRI. AB - This paper discusses the role of a computer advisor in MR image acquisition, interpretation and the diagnosis of cerebral disease. The development of an image and statistical database for use in providing advice is described. PMID- 3043254 TI - MK-801 is neuroprotective in gerbils when administered during the post-ischaemic period. AB - The neuroprotective effects of the non-competitive N-methyl-D-aspartate receptor antagonist (+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine maleate (MK-801) have been evaluated in the gerbil hippocampus when the drug was administered i.p. at various times during and after a 5 min period of transient forebrain ischaemia, induced by bilateral common carotid artery occlusion. A single dose of 1, 3 or 10 mg/kg of MK-801 gave significant protection of hippocampal CA1 and CA2 pyramidal neurons when administered during the occlusion and up to 24 h following the period of ischaemia. A dose of 0.3 mg/kg was effective when administered during the occlusion period but gave no protection at 30 min or 2 h post-ischaemia. Experiments in which MK-801 was administered in repeated doses indicated that significant protection was achieved with 1 mg/kg of MK-801 repeated post-ischaemically and with 1 mg/kg MK-801 supplemented with repeated doses of 0.3 mg/kg of MK-801. However 0.3 mg/kg of MK-801 followed by repeated doses of 0.03 mg/kg administered post-ischaemically was not neuroprotective. These results indicate that MK-801 can protect hippocampal neurons from ischaemia-induced neuronal degeneration when it is administered up to 24 h after the insult. These data provide further evidence that therapeutic intervention in the post-ischaemic period can successfully prevent neurodegenerative events, and that the delayed degeneration of hippocampal neurons following an ischaemic insult occurs by an N-methyl-D-aspartate receptor mediated process. PMID- 3043250 TI - Regulation of protein kinase C activity by various lipids. AB - Protein kinase C has recently attracted considerable attention because of its importance in the control of cell division, cell differentiation, and signal transduction across the cell membrane. The activity of this enzyme is altered by several lipids such as diacylglycerol, free fatty acids, lipoxins, gangliosides, and sulfatides. These lipids may interact with protein kinase C either directly or through calcium ions and produce their regulatory effect (activation or inhibition) on the activities of the enzymes phosphorylated by this kinase. These processes widen our perspective of the regulation of intercellular and intracellular communication. PMID- 3043256 TI - Nerve reconstruction. PMID- 3043255 TI - The free parascapular flap. AB - The parascapular flap is a new, reliable and thin flap with a constant anatomy of the vascular pedicle. The flap can be taken with the lateral border of the scapula or used in combination with the latissimus dorsi muscle. Primary closure of the donor site can be achieved with a flap size up to 30 X 15 cm. The various applications of the parascapular flap are discussed and the results of four patients are presented. PMID- 3043257 TI - Developmental psychopathology: a nine-cell map of the territory. PMID- 3043258 TI - [Defibrotide in the prevention of deep venous thrombosis in general surgery. Preliminary results of a multicenter study]. AB - In an open multicenter comparative study aimed at the evaluation of the efficacy of defibrotide in the prophylaxis of postsurgical deep vein thromboses (DVT) an ad interim evaluation has been made on 2626 patients thus far enrolled. 1323 had received defibrotide (200 mg q.i.d. by IV route from day -1 to day +7th postoperative), 941 calcium heparin (5000 IU b.i.d. or t.i.d. by SC route from day 0 to day +7 postoperative) and 362 other treatments (antiaggregating agents, placebo or no therapy). This group has not been included in the final evaluation, due to its limited size. The diagnosis of DVT or pulmonary embolism (PE) was made according to clinical routinary criteria. The incidence of DTV has been 15/1323 (1.13%) in the defibrotide group and 21/941 (2.23%) in the heparin group (chi square, p = 0.056) while the cases of suspected or ascertained PE have been respectively 3/1323 (0.22%) and 10/941 (1.06%) (p = 0.02). The incidence of adverse effects with defibrotide was less than 1%; occasional cases of increased serum transaminase levels were seen with heparin. These preliminary results supports the effectiveness of defibrotide in the prevention of post-surgery DVT, its effects being similar or more prominent than those of calcium heparin, currently regarded as the standard medication. PMID- 3043260 TI - [Cerebral infarct]. PMID- 3043259 TI - [A sulfadiazine-tetroxoprim combination (co-tetroxazine) in the treatment of the acute exacerbation of chronic bronchitis]. AB - After considering the bacterial flora which is most common in relapses in patients with bronchitis, 40 patients with chronic bronchitis have been treated with tetroxoprim a recently synthetized benzyl pyrimidine associated with sulfadiazine. One 350 mg tablet was administered every 12 hours for different periods, from 7 to 14 days. This study has shown how tetroxoprim has a wide antibacterial range, how it is well tolerated and extremely powerful in treating relapses of chronic infections in bronchi. PMID- 3043261 TI - [Ischemic cerebral vasculopathy in young adults on an angiodysplastic base]. PMID- 3043262 TI - [Cerebrovascular pathology. Comparison of high resolution echography and digital angiography]. PMID- 3043264 TI - [Correlations of computerized tomography aspects and risk factors in patients with RIA (reversible ischemic attack) and stroke. Preliminary results of a multicenter study conducted in Molise]. PMID- 3043263 TI - [B-mode echography in carotid pathology: correlation with morphology]. PMID- 3043265 TI - [Medical therapy of cerebral ischemias]. PMID- 3043266 TI - [Surgical prospectives of acute cerebral ischemia. Present and future]. PMID- 3043267 TI - [Italian Acute Stroke Study: hemodilution + drug. Presentation of the protocol]. PMID- 3043268 TI - [Ticlopidine in the prevention of cerebral ischemia. Experience with a geriatric case load]. PMID- 3043269 TI - [A case of complete thrombosis of the superior sagittal sinus with favorable outcome. Clinical aspects and therapy]. PMID- 3043271 TI - [High-resolution echotomography in the study of supra-aortic branches. Our experience]. PMID- 3043270 TI - [99m-Tc-HM-PAO in the study of cerebral ischemia. Comparison with Doppler echography, digital angiography and computerized axial tomography]. PMID- 3043272 TI - External quality assessment for clinical microbiology in Norway 1986-1987. AB - During the period 1986-1987 eight external quality assessment tests for clinical microbiology were performed, each consisting of four simulated clinical specimens. The results are reported, evaluated and some problem areas discussed. PMID- 3043273 TI - Clinical correlates and diagnostics in carotid vascular disease. PMID- 3043274 TI - Defibrillation flambe. PMID- 3043275 TI - Fever detector strip criticized, defended. PMID- 3043276 TI - Health and disease in Greenland (Kalaallit Nunaat). PMID- 3043277 TI - Cinchona and its alkaloids: 350 years. PMID- 3043279 TI - Advances in the biology and carcinogenesis of basal cell carcinoma. PMID- 3043278 TI - The anencephalic fetus and newborn as organ donors. PMID- 3043280 TI - A view of life and death from a choir loft. PMID- 3043281 TI - On the history of schizophrenia. Evidence of its existence before 1800. PMID- 3043282 TI - Form, fit, and location of the margins of full crowns. PMID- 3043283 TI - A review of the problem of the occlusal vertical dimension of complete dentures. PMID- 3043284 TI - The management of oesophageal varices today. PMID- 3043285 TI - Historical aspects of cervical cancer in New Zealand: concepts and treatment up to 1941. PMID- 3043286 TI - Vesicovaginal fistula revisited. AB - Vesicovaginal fistulas remain common and serious problems for women in West Africa. Thirty-six surgical repairs done during visiting professorships by American gynecologists between the years 1978-1987 are documented. The overall success rate was 70%. The management of large (greater than 4 cm) obstetric fistulas was especially difficult until the Latzko technique was abandoned for the technique of wide mobilization of vaginal flaps over the fistula site. This technique reduced the failure rate from 75% through 1986 to approximately 25% in 1987. Ongoing controversies and basic principles of surgical techniques in fistula repair are reviewed. PMID- 3043287 TI - Current status of genitourinary fistula. AB - From 1970-1985, 303 women with genitourinary fistulas were seen at the Mayo Clinic. The fistula formed after treatment for benign conditions in 74% of the patients and malignant conditions in 14%; in 12%, we were unable to establish the nature of the condition. Gynecologic surgery was responsible for 82% of the fistulas, obstetric procedures for 8%, various forms of irradiation for 6%, and trauma or fulguration for 4%. In the nonirradiated patient, the ideal time for operative repair was eight to 12 weeks after fistula formation or failed repair. With ureterovaginal fistulas, the patient's general condition and the degree of obstruction of the ureter influenced the time and method of repair. We used a vaginal approach for urethral fistulas and an abdominal one for ureteral repairs. Because of difficulty with adequate exposure and the proximity of the ureter, an abdominal approach was used in 20% of the patients with vesicovaginal fistulas; the remaining 80% were approached vaginally, regardless of size, number, or history of previous repairs. Ninety-two percent of the urethrovaginal fistulas were corrected on the first attempt; the four failures were managed successfully at the second attempt. Ninety-eight percent of the vesicovaginal fistulas were corrected on the first attempt when approached vaginally, and all were managed successfully when approached abdominally, regardless of the number, size, or previous operative attempts. PMID- 3043288 TI - The management of persistent clear pelvic cysts diagnosed by ultrasonography. AB - Between December 1981 and June 1986, 52 women examined by ultrasonography were found to have clear pelvic cysts more than 5 cm in average diameter, without septa or solid areas. The cysts were persistent and did not respond to contraceptive pills for four to eight weeks. In all cases, there were histologic or cytologic results from surgical specimens or fluid aspirated from the cysts. In 15 women, the cysts were aspirated and revealed no malignant cells. Thirty seven women underwent laparotomy, of whom 11 (29%) were found to have benign ovarian neoplasms (nine cystadenomas and two dermoid cysts). Because benign ovarian neoplasms are potentially malignant, and because a large number was found in our study, we recommend removal of the cysts by excision rather than aspiration. PMID- 3043289 TI - Clinical trial of naltrexone in premenstrual syndrome. AB - Twenty women with the diagnosis of premenstrual syndrome (PMS) participated in a double-blind, placebo-controlled, crossover study to evaluate the efficacy of naltrexone, an oral opiate antagonist. This study was designed to test the hypothesis that inhibition of opiate withdrawal would aid in the treatment of PMS. The subjects received either placebo or naltrexone from days 9-18 of the cycle for three consecutive cycles. The mean scores of the three day-25 Menstrual Distress Questionnaires of 16 patients on naltrexone were compared with the mean scores of the same patients on placebo. Scores were at least ten points lower on naltrexone in 11 patients and at least ten points higher on naltrexone in two patients. Score changes of less than ten points were noted in the other three patients. The mean scores dropped 28 points on naltrexone (P = .016). The general acceptability of naltrexone was good, with side effects including nausea, decreased appetite, and dizziness. These results suggest that naltrexone alleviates many PMS symptoms and may be an effective treatment for this syndrome. PMID- 3043290 TI - Effect of nicotine on human fetal blood flow. AB - Immediate maternal and fetal cardiovascular responses to different doses of nicotine and carbon monoxide were studied in 24 pregnant smokers. A noninvasive pulsed Doppler ultrasound technique was used for measuring fetal blood flow in the descending thoracic aorta, the intra-abdominal part of the umbilical vein, and in the umbilical artery. Maternal plasma concentrations of nicotine, carbon monoxide, and catecholamines were measured. Maternal heart rate, blood pressure, fetal heart rate, and fetal aortic and umbilical vein blood flow increased, while pulsatility indices of the fetal aortic and umbilical artery blood velocity waveforms decreased, with increasing maternal nicotine levels; all were unaffected by carbon monoxide. Catecholamine levels remained unaffected. These results seem to confirm that the maternal nicotine intake due to smoking has an immediate, dose-dependent effect on fetal blood flow. PMID- 3043291 TI - Pregnancy during residency: II. Obstetric complications. AB - Forty-three percent of women experiencing a pregnancy during a residency education program report medical complications. Analysis of questionnaires from 1197 respondents to a survey of 2000 female physicians indicates that the actual rate of medical and obstetric complications is no different from that in the general population except for the incidence of pregnancy-induced hypertension (12%). Although the incidence was similar for the three specialties studied (obstetrics-gynecology, psychiatry, and surgery), it is higher than that reported in the general population (5%). This may reflect an older maternal age. PMID- 3043293 TI - Pregnancy outcome in women requiring chronic hemodialysis. AB - Chronic renal failure is strongly associated with poor pregnancy outcome. Women dependent upon hemodialysis before conception rarely achieve a successful live birth. Pregnancies in three women requiring chronic hemodialysis before conception and throughout gestation resulted in three live births, of whom one survived the neonatal period. The risks of dialysis in pregnancy (hemodialysis and peritoneal dialysis) are significant to both the mother and fetus. Polyhydramnios appears to be a major complication in women dependent upon chronic hemodialysis. PMID- 3043292 TI - Pregnancy post-Stevens-Johnson syndrome: case report and review of the literature. AB - A pregnancy complicated by vaginal stenosis six years after diagnosis of Stevens Johnson syndrome is described. The pathologic changes in the vagina have not previously been reported. The basic pathology of severe mucosal erythema multiforme was present, as well as ectasia of the superficial capillaries and small venules. The effect of the vaginal scars on the mode of delivery is discussed. PMID- 3043294 TI - Successful twin pregnancy after renal transplant maintained on cyclosporine A immunosuppression. AB - Cyclosporine A has recently been reported to be an effective immunosuppressant agent for use in renal allograft recipients. Questions have been raised regarding its effects during pregnancy, in light of an increased life span and return of fertility in renal transplant patients. A preterm delivery is reported in a cadaveric renal allograft recipient chronically immunosuppressed with cyclosporine A and methylprednisolone. Dizygotic twins were delivered at 35 weeks' gestation, weighing 2452 and 2386 g. Maternal cyclosporine A levels were determined weekly by whole blood radioimmunoassay, with little increase in requirement found before delivery. No indication of maternal renal compromise was apparent, as evidenced by stable weekly creatinine clearance studies. Cyclosporine A, at the doses used, was passed transplacentally, with cord blood values of 34 and 57% of the maternal cyclosporine A level found at delivery. No adverse effects were noted at birth in the average for gestational age neonates, nor at nine-month follow-up evaluation. Given careful monitoring, cyclosporine A may be an effective immunosuppressant agent for use in pregnancies complicated by renal transplantation. PMID- 3043295 TI - Fetal cloacal anomalies: prenatal sonographic findings and differential diagnosis. AB - Serial sonographic findings are presented in three cases of cloacal anomalies. The following sonographic signs were noted: transient fetal ascites, bicystic intra-abdominal structure arising from the fetal pelvis, and oligohydramnios and impaired interval growth. This sequence of sonographic findings may help clarify the natural history of the developing cloacal anomaly: In the early state of its formation, urine enters the abdominal cavity via the fallopian tubes, causing ascites; later, chronic urinary and meconium irritation of tubal mucosa may cause tubal obstruction, which in turn may lead to the formation of genitourinary tract distention, hydronephrosis, and oligohydramnios. Timely diagnosis of cloacal anomalies improves the outcome of pregnancies (delivery in the tertiary care center, early operative correction). PMID- 3043296 TI - Early prenatal diagnosis of familial lipomyelomeningocele. AB - This report describes and illustrates the early prenatal diagnosis of a familial recurrence of congenital lipomyelomeningocele. Diagnosis was made from longitudinal and transverse ultrasound views of the fetal spine at 17 weeks' gestation showing a fourth echogenic area in the lumbosacral dorsal midline. Early diagnosis and resection of this often harmless-appearing, rare teratomatous tumor of the spinal cord is necessary to prevent irreversible neurologic damage during childhood. Familial recurrence has not previously been documented. This case emphasizes that reproductive counseling after the birth of an infant with a rare malformation must always include the possibility of recurrence. The antenatal sonographic and neonatal appearance is presented. PMID- 3043297 TI - Antenatal diagnosis of Pena-Shokeir syndrome (type I) with ultrasonography and magnetic resonance imaging. AB - A case of Pena-Shokeir syndrome type I was diagnosed prenatally with ultrasonography and magnetic resonance imaging (MRI) in a woman with a possible previous occurrence. Initial ultrasonographic examination at 18.5 weeks' gestation demonstrated an unusual appearance of the fetal spine in an otherwise unremarkable fetus. However, subsequent sonographic examinations at 26 and 28.5 weeks demonstrated polyhydramnios and multiple skeletal, brain, and facial abnormalities. Magnetic resonance imaging, performed to further evaluate the fetal brain, confirmed the sonographic findings. However, MRI was not useful in further differentiating the diagnosis. A 1024-g, premature male fetus was delivered at 30 weeks' gestation and died within 30 minutes of delivery. The fetus had multiple congenital anomalies consistent with Pena-Shokeir syndrome type I. PMID- 3043298 TI - Obstetric management of a fetus with nonlethal osteogenesis imperfecta. AB - Osteogenesis imperfecta is the common term for a heterogeneous group of heritable disorders of connective tissue with lethal and nonlethal forms. Prenatal diagnosis presents difficult medical and ethical issues. Of concern are the following: 1) the certainty of diagnosis, 2) the severity of disease, 3) the prognosis for survival and ambulation, and 4) the appropriate mode of delivery. A case of nonlethal fetal osteogenesis imperfecta managed by vaginal delivery is discussed. PMID- 3043299 TI - Sonographic diagnosis of a pregnancy with a diffuse hydatidiform mole and coexistent 46,XX fetus: a case report. AB - Placental molar change with a coexistent live fetus is an unusual entity, particularly when diagnosed in the second trimester of pregnancy. In this case report, the sonographic findings of an abnormally enlarged, diffuse molar placenta with a normal living fetus in the second trimester prompted karyotype analysis. Although triploidy was anticipated, a normal 46,XX chromosomal complement was identified. Histopathology of the placenta after delivery confirmed the rare syndrome of diploid partial mole. Antenatal management of this unusual pregnancy complication is addressed. PMID- 3043300 TI - Fetal bradycardia associated with maternal hypothermia. AB - A case of fetal bradycardia associated with severe maternal hypothermia (92.9F) is reported. Until maternal temperature was corrected, the baseline fetal heart rate (FHR) remained between 90-110 beats per minute without other evidence of fetal distress and despite normal maternal blood pressure and pulse. With rewarming, the FHR gradually returned to normal. Upon return of maternal hypothermia, fetal bradycardia recurred, again responding only to rewarming. This evidence suggests that low maternal temperature alone may lead to alterations of FHR. PMID- 3043302 TI - The conquest of cesarean section-related infections: a progress report. AB - More than a century ago, Robert P. Harris demonstrated convincingly that death from infection after cesarean section could be reduced significantly by operating early, rather than after several days of labor, by using aseptic surgical technique, and by closing the uterine incision. For the most part, his advice was ignored and the mortality rate remained high, except in hospitals with well organized and well-controlled obstetric services. Although the incidence of total infections has been decreased by the use of prophylactic antibiotics, too many serious infections and maternal deaths still occur. These can be reduced by proper management of labor, by recognizing the need for cesarean section early, by using alternative methods for delivery when appropriate, by meticulous surgical technique, and by selective use of prophylactic antibiotics. These changes are not likely to occur unless care of obstetric patients is assumed by experienced obstetricians who are prepared to recognize and correct abnormal labor early and to perform instrumental extraction and vaginal breech deliveries rather than cesarean section in carefully selected patients. PMID- 3043301 TI - Abdominal pregnancy after in vitro fertilization and embryo transfer. AB - Ectopic pregnancy continues to be a major complication of in vitro fertilization (IVF) and embryo transfer. We report the first abdominal pregnancy occurring after this therapeutic approach. The patient, a 35-year-old female, presented a frozen pelvis with a history of severe endometriosis and a left salpingectomy. After the transfer of four concepti in her second IVF/embryo transfer attempt, she became pregnant. Unfortunately, ultrasound evaluation five weeks later showed an ectopic pregnancy in the cul-de-sac. During laparotomy, it was noticed that implantation had taken place near the mesentery of the sigmoid and rectosigmoid. A right cornual tubal ligation was performed. Although the benefit of IVF/embryo transfer far outweighs the risk of an ectopic pregnancy, it is imperative that physicians who care for patients after IVF/embryo transfer be fully aware of the possibility of this complication in this high-risk population. PMID- 3043303 TI - Gastrointestinal complications in gynecologic surgery: a review for the general gynecologist. AB - A working familiarity with the management of common perioperative gastrointestinal complications is required for all general gynecologists. Thermal gastrointestinal injury requires resection of the damaged portion of bowel unless the injury involves only the bowel serosa and is less than 0.5 cm in diameter. Small intraoperative lacerations of the intestine can be closed primarily, whereas larger lacerations often require resection. Some degree of postoperative ileus may be expected, but prolonged ileus requires nasogastric suctioning while excluding bowel obstruction, peritonitis, or electrolyte imbalance. Small-bowel obstruction, most likely to be caused by postoperative adhesions, can often be treated successfully by gastrointestinal intubation. Steps required in the initial management of an enterocutaneous fistula include institution of parenteral nutritional supplementation and antibiotics, skin protection, and investigative studies of the fistula. Preventive measures may be used at the time of any surgical procedure to reduce the incidence of many of these complications. PMID- 3043304 TI - Wolfram's syndrome and HLA. AB - A Sicilian family with three siblings affected by Wolfram's syndrome (Ws) is reported. HLA typing was performed in eight individuals from this family through three generations. Two of the three patients were HLA DR2 positive. The results suggest that the gene for Ws is not linked to the HLA region on chromosome 6, but located on some other chromosome, and that the allele HLA DR2 might predispose to the mutation responsible for Ws. PMID- 3043305 TI - Early oral changes following bone marrow transplantation. AB - This study assessed and analyzed the early oral changes following chemoradiotherapy and bone marrow transplantation. The most notable changes involved mucosal color (white and red), atrophy, vascularity, ulceration, increased salivary viscosity and xerostomia, and the patients' subjective complaints of dryness and oral pain. The ventral tongue, buccal and labial mucosa, and marginal gingiva manifested the most notable changes, while the palate was least affected. The overall trend was for the oral changes to begin slightly before transplantation, to worsen over the first 2 weeks after transplantation, and then to resolve progressively over the remainder of the study period. These oral changes appear to result from a number of insults, including the conditioning chemoradiotherapy, posttransplant immunosuppressive chemotherapy, xerostomia, local trauma, oral infections (especially those caused by HSV), and possibly acute GVHD. Oral HSV infection and/or acute GVHD should especially be considered if the oral status markedly worsens 21 days or more after transplant. PMID- 3043306 TI - Arteriovenous malformation of the maxillary sinus: an unusual clinical presentation. AB - Arteriovenous (AV) malformation is a rare vascular lesion of the jaws with a benign clinical presentation. As a result, AV malformation may be excluded from a presurgical differential diagnosis, with catastrophic results. Review of the pertinent literature showed no documentation of AV malformation in the maxillary sinus, and therefore we present the case of a 43-year-old white man with a history of blunt trauma to the paranasal region, whose differential diagnosis included a vascular lesion. PMID- 3043307 TI - Ingestion of a unilateral removable partial denture causing serious complications. AB - A case of ingestion of a unilateral removable partial denture is presented. Routine endoscopic retrieval was unsuccessful, and the patient required surgical removal of the prosthesis. Subsequent hospitalization, complications, and recovery from an apparently simple accident are discussed. PMID- 3043308 TI - Transplantation and implantation procedures after surgical treatment of ameloblastoma. AB - Attention is called to the fact that in cases of large ameloblastomas systematic, careful treatment is essential. This implies proper reconstruction of the jaw and restoration of the occlusion after adequate extirpation of the tumor. A case involving a large mandibular ameloblastoma is reported. It was successfully treated, step by step, by tumor extirpation, jaw reconstruction with rib transplants and installation of titanium implants (Biotes) in the rib transplants, anterior vestibuloplasty with skin graft, and, finally, an overdenture. Carefully planned coordination of well-documented surgical and prosthetic procedures has achieved a functionally and esthetically successful result. PMID- 3043309 TI - Myasthenia gravis as a cause of facial pain. AB - A case of myasthenia gravis is described. The diagnosis was confirmed electromyographically and immunologically; the presenting features were those of myofascial pain. Attention is drawn to the association of myofascial pain with other systemic diseases and stresses the need to look critically at the patient for evidence of a systemic origin for myofascial pain. PMID- 3043310 TI - Ameloblastic fibrosarcoma. AB - Ameloblastic fibrosarcoma is an extremely rare tumor. To date only 43 cases have been reported in the literature. An additional case of ameloblastic fibrosarcoma is presented; the clinical features, histologic characteristics, treatment, and the relevance of the presence of dental hard tissue are discussed. PMID- 3043311 TI - [Extra-articular elbow fractures]. AB - Only some fractures of the humerus-shaft or the shaft of radius and ulna can be called extra-articular fractures of the elbow. These fractures occur mostly in the still growing bone or in bones with abnormal structure. The treatment consists in adults in open reduction and osteosynthesis, in adolescents and children percutaneous methods of fixation can be recommended. Conservative and early functional treatment are only indicated in very few cases. Lesions of nerves and vessels are relatively common. PMID- 3043313 TI - [Conservative treatment of elbow dislocations]. AB - Ligamentous injuries following dislocations of the elbow joint are a primary indication for nonsurgical treatment. The principles of closed reduction are longitudinal traction of the forearm, followed by flexion. The period of immobilization in a plaster cast should be decided on an individual basis. For elbows that show no tendency to redislocate, immobilization for no more than 1 week should suffice. For elbows with a tendency to redislocate 2-3 weeks' immobilization is recommended. Radiological follow-up shows a high percentage of degenerative changes, i.e. subchondral sclerosis, osteophytosis, periarticular calcification, irregularities in the epicondyles. In addition to slight degenerative changes, there are few subjective complaints and functional impairments. Now evidence has been found to recommend primary surgical treatment of ligamentous injuries associated with dislocation of the elbow. PMID- 3043312 TI - [Indications for operation in elbow dislocation]. AB - This paper briefly presents the anatomy of the elbow joint, its pathophysiology, the various types of dislocation of the elbow joint and concomitant injuries, the appropriate diagnostic procedures and the indications for surgery and the operative technique applied for correction of dislocation. The only urgent indications for operative treatment are elbow joint dislocation with concomitant bone injuries, persistent instability or luxation position, open injuries and vessel and/or nerve injuries. For purely ligamentous lesions combined with relative loss of stability an operative procedure does not seem necessary. The operative technique applied for the treatment of habitual or recurrent dislocation is also described. PMID- 3043314 TI - [Surgical differential therapy of fracture of the radius head]. AB - The different types of radial head fractures require specific and varying methods of treatment. Between 1982 and 1985, follow-up evaluation of 92 out of 110 patients treated for such fractures at our facility could be performed. The most frequently seen fractures: nondisplaced marginal and marginal sector fractures, as well as fractures of the radial neck, for example, can be treated conservatively. Follow-up results of such treatment were mostly good to very good. The minimal depression injury is also well suited for conservative therapy. Opened reduction and internal fixation is indicated only in cases of extreme depression of the radial head. Displaced marginal, marginal sector or neck fractures exhibiting a tilting of more than 20 degrees require osteosynthesis. The limitations of internal fixation are encountered in reconstruction attempts of comminuted radial head fractures. Due to the poor results obtained after such attempts, internal fixation of such injuries cannot be recommended. Comminuted fractures and neck fractures with an extensive comminuted region as well as displacement of the radial neck more than 20 degrees, require resection, if active elbow movement is not to be expected within 3 weeks. If resection of the radial head cannot be avoided, it should be performed within the first 5 weeks post-injury. Results following early resection are markedly better than those achieved with late resection. PMID- 3043315 TI - [Monteggia injuries]. AB - Sixty-three recent Monteggia lesions are reported, which were all treated operatively. In 52 patients, the ulna was stabilized by a compression plate. Dislocation of the radial head was reduced by a closed procedure in 24 patients and 38 patients an open operation was performed (including fractures, sutures of the annular ligament, and resections of the radial head). After 38 months, the results were better in comparison to series using conservative or medullary pinning (88% good and very good). Poor results must be expected if the operation is delayed for 2 or more weeks, if the radial head has to be resected, or in posterior Monteggia lesions. The late reconstruction of a neglected Monteggia lesion by means of an osteotomy of the ulna shaft and plate fixation is described in a child. PMID- 3043316 TI - [Experiences with the preventive use of aminophylline (Euphylline) in apnea and periodic respiration in infants]. AB - Sleep apnea and periodic breathing in infants are the result of a mild hypoxia and they are the requirement for SIDS. As a results of the modern medicine each 4.-5. death is the cause of SIDS during the first year of life. The positive effect of theophylline on apnea of prematurity is known since 1973. We have given theophylline (Euphyllin) from 1979 till 1986 to 198 premature infants with a birth weight below 2000 g for apnea prophylaxis orally in a doses of 9 mg/kg body weight per day distributed in 3 to 4 doses for a period of 1 to 4 months. Since then primary apnea in premature infants haven't any importance. Beside the home monitoring we give theophylline for SIDS. In 1986 and 1987 1041 healthy term newborn infants received theophylline over a period of 6-8 weeks and more then 300 newborns until 6 months and more. There were no deaths from these infants form the cause of SIDS. Since 1987 all newborn infants with a pathological hypoxia-test as a screening test for the risk of SIDS received Euphyllin until normalization. With this method we have reduced evidently the death rate of SIDS in our district. PMID- 3043317 TI - Insulin regulatory effects on purine- and pyrimidine metabolism in alloxan diabetic rat liver. AB - Aim of this study was to elucidate insulin regulatory effects on purine and pyrimidine metabolism. Livers of alloxan diabetic and insulin treated rats were freeze clamped and nucleotide pools measured using HPLC techniques. Activities of key enzymes of de novo and salvage pathways were analyzed with radioassays. In diabetic liver nucleotide triphosphate pools were reduced between 46 and 75% of controls, nucleotide monophosphate concentrations increased. Activities of de novo biosynthetic enzymes amidophosphoribosyltransferase, FGAM synthase, IMP dehydrogenase, GMP synthase, carbamoylphosphate synthase II were curtailed by 16 61%, those of salvage enzymes hypoxanthine-guanine-phosphoribosyltransferase, adenine-phosphoribosyltransferase, thymidine kinase also decreased to 31-58%. Insulin treatment for 2 and 7 days normalized nucleotide pools, activities of key enzymes of de novo and salvage pathways were increased between 2.4 and 4.1 fold compared to diabetic untreated. Activation of nucleic acid metabolism by insulin can be explained by the requirement for high energy phosphates of certain anabolic key enzymes in carbohydrate and lipid metabolism. Impaired synthesis in insulin deficiency of end products of guanylate and pyrimidine pathway required as substrates for a variety of enzymes synthesising membrane structures throw new light on the pathogenesis of some late complications of diabetic disease. PMID- 3043318 TI - Neuroendocrine evaluation of central opiate activity in primary headache disorders. AB - The evaluation of central opiate activity could be of clinical value in the diagnosis and treatment of pain syndromes. The current approach via direct measurement of endogenous opioid peptides in cerebrospinal fluid (CSF) is not devoid of side effects and cannot be used in every-day practice. As an alternative to this method, we have studied the neuroendocrine response of plasma LH to an i.v. naloxone injection in 39 headache sufferers from different diagnostic subgroups, and in 12 age- and sex-matched healthy volunteers. Patients (19 females and 20 males) were affected by common migraine (CM, 11 cases), migraine with interparoxysmal headache (MIH, 9), classical migraine (CIM, 9), and chronic cluster headache (CH, 10). Headache lasted 3-36 years. Prior to naloxone challenge (4 mg i.v.), LH pulsatility was evaluated for 1 h. The next morning, the pituitary response to LH-RH (10 micrograms i.v.) was tested in 20 patients. Plasma LH was measured by RIA in every sample. The response to the tests was evaluated as secretion area of plasma LH minus the mean basal value. Controls (497.5 +/- 85.5 mIU/ml x 120 min), ClM (357.8 +/- 78.9) and CH (450.5 +/- 70.4) patients showed similar results, while in cases of CM (155.3 +/- 71.7, P less than 0.05) and MIH (104.1 +/- 53.7, P less than 0.01) the LH secretion after naloxone injection was significantly blunted. On the contrary, the response of LH to LH-RH was similar in controls and patient groups, thus excluding pituitary dysfunctions in this response.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3043319 TI - Special issue dedicated to Arthur J. Helfet, M.D. PMID- 3043320 TI - Arthur J. Helfet. PMID- 3043321 TI - Osteoarthritis. AB - Osteoarthritis (OA) is one of the most common chronic diseases and the most frequent cause of rheumatic complaints in the older population. While still regarded primarily as an expression of a degenerative disease of cartilage accompanying the aging process, newer concepts in many areas have emerged. In particular, these concepts relate to cellular interactions among joint components, biochemistry and ultrastructure of cartilage, biomechanics, pharmacology and antiarthritic drugs, surgical approaches to the repair of arthritic joints, and population studies. However, it is still very difficult at this time to put together a coherent picture of the evolution of OA, and this profoundly limits our understanding of the disease process. In population studies we often rely on physical inspection of joints or complaints of pain, both of which may be of limited value despite extensive damage to the articular cartilage. This makes us dependent on x-rays for case finding, and these are costly and difficult to do. Longitudinal studies are not feasible because we are talking about a survey that should extend for several decades. We have limited access to samples of joint components for biochemical studies in the 20 to 40 year-old population, although surgical intervention for menisceal injury should offer an opportunity to observe conditions where the evolution of OA is often accelerated. This discussion, written by an internist, is an attempt to survey aspects of the present state of our knowledge, albeit incomplete, of OA involving the joints of the extremities. It is hoped that orthopaedic surgeons in particular will be stimulated to explore further the evolution of OA, an area to which they have contributed so significantly. PMID- 3043323 TI - A vignette of personal experiences in World War II. PMID- 3043322 TI - Chondrosarcoma of the small bones of the hand: case report and review of the literature. AB - An unusually rare case of chondrosarcoma arising from an osteochondroma of a finger is presented. Chondrosarcomas of the hand in general are rare and almost always central in origin. Malignant degeneration of previously known enchondromas can occur and account for some but not all cases of chondrosarcoma of the hand. Histologic interpretation of cartilagenous lesions of the hand is difficult, and clinical behavior and radiographic appearance and appearance at surgery are often more reliable indicators of malignancy. Clinical suspicion should be aroused in an older individual with a previously quiescent lesion that becomes larger and/or more symptomatic. The clinical course of these tumors is slow and metastasis is late and usually to the lungs. Treatment of choice is amputation or ray resection of the affected part. Prognosis is good if metastasis has not occurred. PMID- 3043324 TI - A portrait of Dr. Arthur J. Helfet. PMID- 3043325 TI - The placement and treatment of thoracolumbar spine fractures. An algorithmic approach. AB - Advances in imaging techniques and surgical instrumentation systems have made the selection of appropriate treatment for thoracolumbar fractures difficult. A review of the literature and personal cases have led to the development of a series of algorithms for patient care. Fracture patterns with recommended therapeutic approaches are presented and discussed. PMID- 3043326 TI - Volkmann's ischemic contracture. A case report. AB - A case is presented illustrating some of the basic principles in the development and repair of Volkmann's ischemic contracture following a supracondylar fracture. The etiology, initial signs, prevention, and possible treatments of the contracture are discussed. Early fasciotomy is the best treatment for impending contracture, while neurolysis with infarct excision, a flexor pronator slide, and tendon transfer can return much function following established contracture of the forearm. PMID- 3043327 TI - Interactions between malaria parasites infecting the same vertebrate host. AB - Several species of malarial protozoans commonly parasitize the same host population and often the same individual host. This paper reviews the evidence for interactions among such host-sharing parasites. Field studies measuring the cross-sectional prevalence of malarial species often record fewer mixed infections than expected by chance, suggesting that one parasite has excluded another or suppressed its parasitaemia to undetectable levels. Prevalences may vary reciprocally between seasons, with one species increasing in prevalence while another decreases, despite parallel increases in the transmission rates of both, again suggesting suppression of one species by another. However, longitudinal studies of individual hosts indicate that malarial parasites may also favourably affect the host environment for each other, perhaps due to their depressive effect on the immune system: this is shown by the recrudescence of a latent malarial species immediately before or after the parasitic wave of another species. The suppression hypothesis is supported by data derived from the simultaneous inoculation of two Plasmodium species into laboratory animals; many studies have shown that one or both species are suppressed. This may be mediated by competition for host cells or nutrients, or by heterologous immunity. However, the suppressed species rebounds after the other species has abated, and may show a prolonged infection. Experimental evidence that one species can facilitate the recrudescence of another is minimal, but this may reflect the paucity of investigations of this phenomenon. Laboratory studies show only minor cross resistance between host-sharing species, which is consistent with the hypothesis that their co-occurrence has led to antigenic divergence or that species showing strong heterologous resistance cannot co-exist in the same host population. Such complementarity occurs not only with the host immune response but also with many other life-history characteristics of host-sharing parasites, such as host cell preference. I conclude that malarial species have been important in each other's evolution, particularly in the tropics where multi-species complexes are common. PMID- 3043328 TI - [Interdisciplinary meeting on anti-infectious chemotherapy. 3 December 1987, Paris. Proceedings. 1]. PMID- 3043329 TI - [In vitro bactericidal effect of cefotetan-aminoside combinations]. AB - Bactericidal activity of cefotetan-gentamicin combinations was studied on 6 bacterial strains: S. aureus, K. pneumoniae, E. cloacae, S. marcescens, P. vulgaris, P. stuarti. Time kille curves technic was performed with final concentrations of cefotetan: 4 at 32 mg/l, and of gentamicin: 0.25; 0.5; 1; 2; 8 mg/l and with an 10(6) CFU/ml inoculum. Cefotetan at 4 mg/l was not able to obtain a 0.01% (percentage of survivors) bactericidal activity before 24 h. The combination cefotetan-gentamicin (0.25 to 2 mg/l according to the strains) were bactericidal (0.01% of survivors) before 24 h: 1 to 6 h according to the strains, more rapidly than with gentamicin alone at the same concentration. This more rapid bactericidal activity obtained by cefotetan-gentamicin combination seems to indicate this combination in the treatment of severe infections in immunocompromised patients. PMID- 3043330 TI - [Titration curves (ph gradient electrophoresis) of SHV-1 and SHV-2 beta lactamases and a new type]. AB - The molecular structures of the SHV-1 (p 453) and SHV-2 (pBP 60-1) beta lactamases and of a new enzyme, a SHV-2 like extended broad-spectrum beta lactamase (86-4), were compared by analysis of their titration curves (pH gradient electrophoresis). The titration curves of SHV-1 and SHV-2, which have the same isoelectric points (pI = 7.7). were completely superimposable for the whole of the pH gradient (pH 3.5-10), indicating a close homology between the two proteins, with perhaps the substitution of several amino acids by ones having the same charge. The curves of SHV-1 (pI = 7.7) and the new SHV-2-like enzyme (pI = 6.98) indicated that a basic residue in SHV-1 has been replaced by an acidic residue in the new SHV-2-like enzyme. These results show that, like SHV-2, the new beta-lactamase is a variant of SHV-1, and that the structural differences are probably limited to a very small number of amino acid residues. Nevertheless, this new beta-lactamase (SHV-3) may have arisen directly from SHV-1, indirectly via SHV-2, or even from another beta-lactamase. PMID- 3043331 TI - [In vitro antibacterial activity of a new macrolide: miokamycin. Comparison with erythromycin and josamycin]. AB - Minimal inhibitory concentrations (MIC) of miokamycin (M) were evaluated by agar dilution, comparatively with erythromycin (E) and josamycin (J) for 81 bacterial strains chosen as a function of susceptibility and resistance to Macrolides Lincosamides-Streptogramins group (MLS). On Gram positive cocci, mode MIC of E, J, and M for strains sensitive to MLS were respectively (micrograms/ml): Staphylococci: 0.25; 1; 2-Streptococci and Pneumococci: 0.016; 0.03-0.12; 0.06 0.25-Enterococci: 0.5; 0.5-1; 1-2. Activity of the three antibiotics was similar against Staphylococci resistant to lincomycin and streptogramin A, those resistant to streptogramins A and B, on Staphylococci and Streptococci only resistant to lincosamides by inactivation. M, as J, was active on coagulase negative (Staphylococci resistant to E by inactivation and on MLBB inducible Staphylococci; on these strains, M is a resistance non-inducible macrolide. Activity of E, J and M was reduced on MLSB inducible Streptococci. The three antibiotics were inactive on Staphylococci Streptococci and Enterococci MLSB resistant constitutive. On Haemophilus, E (2-8 micrograms/ml) was more active than J (2-16) and M (8-32). Thus, M, as J, was shown to be among macrolide antibiotics of resistance non-inducing type on MLSB inducible resistant Staphylococci; its activity was slightly inferior to that of J, but very similar to that of spiramycin, other macrolide of this group. PMID- 3043332 TI - [An expert system as an aid to the validation of results of the antibiogram. Feasibility study based on the example of Staphylococcus aureus]. AB - A model of expert system using Prolog language was developed, to verify the coherence of the results of the antibiotic sensitivity test. Biological knowledge was formalized in three different ways: a credibility coefficient based on epidemiological data is assigned to known observed resistance; co-existent resistances are described with lists of "implicit" resistances, reflecting phenotypes commonly observed within some antibiotic groups; every single or "implicit" resistance is connected to a "gregarius" status, expressing the plasmidic nature of resistance. Applied to Staphylococcus aureus, the expert system is able to detect the inconsistencies of the antibiotic sensitivity test and to identify required knowledges. It therefore allows phenotypic interpretation of results. PMID- 3043333 TI - [Detection of heterogeneous Staphylococci by the Autobac system using a nafcillin disk]. AB - Detection of Methicillin Resistant Staphylococcus aureus (MRSA) is still a problem for automated system users. Forty-nine strains were studied--two kinds of tests were performed: the first by using a 1.0 microgram nafcillin disk with the Autobac system, the second by testing an oxacillin disk by diffusion on Mueller Hinton agar base complemented with natrium chloride incubated at 37 degrees C. Between the two tests, we observed a concordance for 87.8% of the strains. Six strains showed a discrepancy with the reference method. But for five of them, the Light Scattering Index (LSI) showed a decreasing sensibility near for the cut off. Whereas, only one strain is considered as susceptible. The 1.0 micrograms nafcillin disk allow better results than the 5.0 micrograms oxacillin disk in the automated system, Autobac. PMID- 3043334 TI - [In vivo bactericidal activity of an oxacillin-netilmicin combination against Staphylococcus aureus. Influence of the rhythm of netilmicin administration]. AB - The bactericidal activity of two regimens of netilmicin (8 mg/kg/day) given intravenously once a day (od) or thrice daily (tid) both alone and in combination with oxacillin (200 mg/kg/day) was compared using a model of fibrin clots infected with a strain of Staphylococcus aureus (10(7) CFU/g) sensitive to methicillin and netilmicin (clinical isolate) and implanted subcutaneously in rabbit. This study shows that: 1) Netilmicin given alone as both single and divided doses results in early bacterial killing but does not exert a bactericidal effect after 24 hours because of a significant late increase of the number of bacterial. 2) The netilmicin-oxacillin combinations are more bactericidal at 1 h, 2 h and 24 h than oxacillin alone (P less than 0.001). 3) The oxacillin-netilmicin combination appears to be better for bacterial killing when netilmicin is given thrice daily (P less than 0.001). It is hard to draw a clinical inference from such an experimental study but it seems that 8-hour divided doses intervals should be better for administration of netilmicin than single daily dose during the acute period of staphylococcal infections. PMID- 3043335 TI - [Sensitivity to antibiotics of 64 strains of Stomatococcus mucilaginosus isolated in human clinical cases. Demonstration of erythromycin resistance]. AB - Susceptibility to fifteen antibiotics of 64 stains of Stomatococcus mucilaginosus isolated from bronchial secretions was tested by susceptibility antibiotics diffusion and MIC method. Sixteen phenotypic profiles were determined by MIC. Some of these profiles and the spontaneous loss of resistance to erythromycin indicate plasmidic origin of this resistance. PMID- 3043336 TI - [Value of bead-type cassettes for antibiograms of Streptococci in Autobac units]. AB - Antimicrobial susceptibility testing of Streptococci in automated units (Autobac) is not always satisfactory: aggregates formed spontaneously by Streptococci in the medium used can reduce the reliability of tests by interfering with correct reading, and, in many cases, bacterial growth is very slow, necessitating long periods for analysis (up to 6 hours). Owing to the fact that use of "bead type" cassettes for antifungal susceptibility testing in the Autobac improved results, and in particular prevented the formation of aggregates, we decided to adopt the same system for tests of streptococci. Each compartment in these cassettes contains glass microbeads which prevent aggregate formation thanks to an agitation mechanism. Rapid sedimentation of these beads allows normal reading with the Autobac. A comparative study of 66 strains of streptococci with various antibiotics was performed. Three techniques were used: classical diffusion technique, susceptibility testing with the standard cassette in the Autobac, and test with the bead type cassette in the Autobac. Parameters evaluated included the effective duration of each test, and comparison of the results (Autobac with and without beads, diffusion method). Results are detailed and discussed. PMID- 3043337 TI - [Kinetics of ofloxacin-amoxicillin and ofloxacin-clavulanic acid combinations against pathogenic bacteria of the respiratory tract]. AB - Ofloxacin exhibit a good activity against the pathogen bacteria of the respiratory tract, except for S. pneumoniae. Its use is interesting because it has a good diffusion into respiratory tissues. However, the combination with an another antibiotic was necessary to spread the activity spectrum and to prevent the resistant variants. By the new killing curve checkerboard method, these 2 combinations are studied against 8 strains: 4 H. influenzae (2 beta-lactamase producers), 2 S. pneumoniae and 2 K. pneumoniae, at 0, 2, 4, 6 and 24 hours, a viable bacteria counting is executed by a microdilution method and is subcultured with a Steers-type multiple inoculator. With K. pneumoniae, ofloxacin has a dose dependent bactericidal activity which is maximum at 1 mg/l, While augmentin has a time-dependent activity. In the combination the synergy is rare. With H. influenzae, the results are the same; the bactericidal activity is less rapid but it is better than the ones with amoxicillin and clavulanic acid. With S. pneumoniae, the 2 antibiotics have the same activity. No antagonism is observed. And the antibiotic which has a better bactericidal activity determine the viable bacteria count. PMID- 3043338 TI - [Activity of 9 beta-lactam antibiotics combined with clavulanic acid or sulbactam against the strains of broad-spectrum beta-lactamase (CTX-1) producing Enterobacteriaceae isolated at the Henri Mondor Hospital]. AB - Minimal inhibitory concentrations (MICs) of seven cephalosporins: cefotaxime (CTX), ceftriaxone (CRO), ceftazidime (CAZ), latamoxef (MOX), cefoxitin (FOX), cefotetan (CTT) and CM 40876 (CM), of aztreonam (ATM) and imipenem (IPM) were evaluated by agar dilution with and without 5 mg/l of clavulanic acid (AC) or sulbactam (SB) for 28 strains isolated in 1986 (15 K. pneumoniae, 3. K. oxytoca, 4 E. coli, 4 E. cloacae, 1 E. aerogenes and 1 C. freundii). Comparatively to MICs of sensitive strains and to those of cured variants, MICs of these strains were very increased for CTX, CRO, ATM (mode MIC: 1 mg/l), and CAZ (2); weakly increased for MOX and CTT (0.25), and identical for IMP (0.12-0.25), CM (0.06) and FOX (2-4), except for Enterobacter and Citrobacter (64). Association with AC or SB did not modify MICs of FOX, CM and IMP. For the other antibiotics, MICs were reduced by addition of AC: Klebsiella: 5 log2 for CTX and CRO, 4 for CAZ and ATM, 2 for MOX and CTT; E. coli: 4 log2 for CTX and ATM, 3 for CRO and CAZ, 1 for MOX and CTT; Enterobacter and Citrobacter 2 log2 for CTX, CRO, CAZ and ATM, 1 for MOX and CTT. With SB, decrease of MICs was two to for fold lesser than with AC. AC, and less efficiently SB, restored activity of CTX, CRO, CAZ and ATM on CTX-1 producing Enterobacteriaceae, particularly Klebsiella and E. coli. It was the same for MOX and CTT, weakly affected by this resistance. AC and SB had not effect on FOX, CM and IPM which remained active on these strains. PMID- 3043339 TI - [Current status of aminoglycoside resistance in hospital Enterobacteriaceae]. AB - We studied the susceptibility of Enterobacteriaceae to four aminoglycosides (gentamicin, tobramycin, netilmicin et amikacin). We determined their phenotypes of resistance by taking into account both the critic concentrations of the CFA (french committee for antibiogram) and the MIC of the main susceptible population of each species. The most frequent phenotypes were GTNt, TNtA and GTNtA. Amikacin resistance including phenotypes were essentially found in Klebsiella and Serratia (35% and 53% of the strains, respectively); with respect to amikacin, the phenotype expression may be insufficient to exceed the sensitive critic concentration of the CFA. Amikacin resistant strains were isolated from chronically infected patients with devices, such as urinary catheters or tracheal cannula. These results suggest a strains or plasmids outbreak. PMID- 3043340 TI - [Sensitivity to clavulanic acid of Escherichia coli isolated at the Henri Mondor hospital in 1986 according to phenotypes of beta-lactam antibiotic resistance]. AB - Susceptibility to Augmentin (AUG) was studied as a function of resistance phenotypes toward amoxicillin (AMX), carbenicillin (CAR), cephalothin (CFT) and cefotaxime (CTX) for 1817 strains of Escherichia coli isolated at Henri-Mondor hospital during 1986. For strains susceptible to the 4 beta-lactams (phenotype SSSS: 66%), the median zone diameter observed with AUG was 26.4 mm. It was slightly inferior (22.1) for acquired - penicillinase producing strains (phenotype RRSS: 21.5%), but zone diameters were greater than or equal to 21 mm for over than 75% of these strains. Strains of phenotype RRIS (6%), probably high penicillinase-producers, were generally intermediate to AUG (median zone diameter: 17.8), showing partial inhibition of enzyme by clavulanic acid. Cephalosporinase hyperproducers mutants (phenotype RSRS: 4%) and strains with a RRRS phenotype (2.5%), probably penicillinase and cephalosporinase producers, were resistant to AUG (median zone diameters: 15.6 and 12.6 mm). Among rarely observed phenotypes, some were readily integrated to major phenotypes: RSIS, ISRS and ISIS to phenotype RSRS; RIRS to phenotype RRRS; IISS to phenotype RRSS; apparently "aberrants" phenotypes (ISSS, RSSS, SSIS, SSRS) required interpretation of results as a function of observed zone diameters and at time a verification of tests. This study confirms the in vitro activity of AUG on penicillinase producing E. coli; in addition simultaneous reading of zone diameters observed with AMX, CAR, CFT and CTX, permitted to infer the probable mechanism of resistance to beta-lactams and therefore to correct possible discrepancies observed in antibiogram results. PMID- 3043341 TI - [A mathematical study of the mortality curve of Escherichia coli exposed to aminoglycosides]. AB - The time killing curves of three strains of E. coli (CIP54117, ATCC25922 and ATCC29194) exposed to kanamycin, amikacin, netilmicin and dibekacin are decreasing exponentials. The absolute value of the killing rate m is related to the antibiotic concentration through a Michaelis-Henry equation. A maximum killing rate m max and an affinity constant Kc between bacteria and antibiotic can thus be estimated. m max is mainly strain dependent. On the contrary, Kc is related to the antibiotic. Kc is much higher (i.e. the affinity is much lower) for kanamycin and amikacin than for netilmicin or dibekacin. The dose-effect curve is a saturation curve. Increasing doses of antibiotic do not increase the mortality proportionally. PMID- 3043342 TI - [Inhibition of bacterial adhesion of uropathogenic Escherichia coli strains by the urine of patients treated with nitroxoline]. AB - The present study evaluates the effects of sub-inhibitory concentrations of nitroxoline, (oxyquinoline derivative) widely used in the treatment of uncomplicated, urinary tract infections, on the adherence of uropathogenic strains of Escherichia coli. These bacterial strains showed mannose sensitive and/or mannose resistant hemagglutinating activity (HA). In the presence of nitroxoline and at sub-MIC concentrations, inhibition of adherence is 90% (MIC/4), 87% (MIC/8), and 70% (MIC/16), whatever HA's are expressed by the E. coli strains. The inhibitory effect on adherence is also observed in the urine after oral administration of 400 mg of nitroxoline. The concentrations of nitroxoline in the urine are determined by microbiological assay (anti-bacterial activity) and by physico-chemical assay (total nitroxoline and free nitroxoline). The percentages of inhibition are related to the concentrations of free and conjugated nitroxoline. For a 1/16 dilution of urine, the inhibitory effect is 70% and 87% respectively 1 h 30 and 2 h 30 after oral administration of nitroxoline. After 5 h, a similar inhibitory effect is observed for a 1/2 dilution of urine. These results justify the performance of a clinical trial on the prophylaxis of recurrent urinary tract infections by nitroxoline. PMID- 3043343 TI - [Action of imipenem on Enterobacter cloacae]. AB - Most of Enterobacter cloacae strains produce chromosomally determined class I beta-lactamases when they are exposed to beta-lactams. Imipenem is a strong inducer of these enzymes but is poorly affected by them. We compared the effect of imipenem on inducible, non-inducible and stably derepressed strains of E. cloacae using the killing curve system. With antibiotic concentrations of 0.5 mg/l or more, an intense dose-dependent bactericidal effect was observed within 4 to 6 hours. However the bactericidal activity was incomplete. With an inoculum as low as 10(5) bacteria/ml a regrowth was observed after 24 hours. Among the surviving bacteria imipenem had the same bactericidal kinetic than on the parental strain. Similar results were observed with inducible, non-inducible and stably derepressed strains. PMID- 3043344 TI - [In vivo study of the sensitivity of Treponema pallidum to ofloxacin]. AB - Treponema pallidum has not been yet cultivated. Hence any in vitro investigation is excluded, and it is owing to the experimental animal model, the rabbit, that we have studied the susceptibility of that germ to ofloxacin. This quinolone, owing to its pharmacokinetic and therapeutic properties, can specially be indicated in the treatment of Sexually Transmitted Diseases. Thus, its appeared to be of the utmost importance to know if the suggested schedule of treatment for STD, might not be susceptible to modify the course of a co-existing incubating syphilis by either delaying or inhibiting the apparition of the clinical features of primary syphilis. This study was undertaken at the incubation period, in syphilitic rabbits, using kinetic data obtained in man, after a given dosage of ofloxacin. Results were appraised upon converging data: lesions, bacteriology, and serology of the tested lot compared with two control batches of infected rabbits, the first one being untreated, the other having received the reference antibiotic treatment. From the data obtained and in the experimental settled conditions where this study was done, it results that ofloxacin has no effect on the course of the experimental syphilitic infection. PMID- 3043345 TI - [In vitro activity of an ofloxacin-metronidazole combination against anaerobic bacteria. Kinetics of the action of metronidazole against Bacteroides fragilis]. AB - The activity of metronidazole combined with ofloxacin was investigated by the checkerboard method in liquid medium against 60 obligate anaerobes. The bacteriostatic effect of the combination was assessed by calculating the FIC index. Two metronidazole resistant strains of Propionibacterium acnes (MIC greater than 32 mg/l) were inhibited by 0.125 mg/l of this former antibiotic in presence of an ofloxacin concentration equal to half the value of the MIC. On the 58 other anaerobic strains, the combination of metronidazole plus ofloxacin had an additive bacteriostatic effect (30 strains) or a synergistic effect (26 strains). No antagonism was noted with any strain. For selected anaerobic or mixed infections the combination of metronidazole and ofloxacin may be useful. Killing curves demonstrated that, under good anaerobic conditions, metronidazole acted rapidly against Bacteroides fragilis. At a concentration of 8 mg/l, a decrease of log10 of the population was observed after 2 hours. After a contact ranging from 1 to 8 hours, depending of the investigated strains, a bactericidal effect was observed with metronidazole. PMID- 3043346 TI - [Rickettsiaceae infections and fluoroquinolones]. AB - Twenty nine patients with Rickettsiosis (Tick born fever, n: 22, Q fever, n: 7) has been treated with fluoroquinolones. These compounds was used alone n: 25 (Pefloxacin, n: 15; Ofloxacin, n: 10) or associated n: 4 (Pefloxacin + Rifampin). The general efficacy was excellent. Common Tick born fever and pulmonary forms of Q fever was fast cured. Severe Tick born fever (neurologic or polyvisceral forms, n: 5) and Q fever with long term hyperthermia, n: 4 cas always cured without complications but the time necessary to obtain apyrexia was often longer (6 cases). PMID- 3043348 TI - [Pharmacokinetics of piperacillin during continuous ambulatory peritoneal dialysis]. AB - We studied the kinetics of piperacillin in patients under continuous ambulatory peritoneal dialysis. Piperacillin 2 g was injected intravenously in 6 patients whereas 1 g was given intraperitoneally either a single dose in 3 patients without infection or the same dose every six-hours in 4 patients with peritonitis. Piperacillin was assayed by HPLC. After intravenous administration, the mean plasma piperacillin concentration was 3.1 +/- 5.6 mg/l at 12 h, the mean plasma t1/2 at 2.43 +/- 0.84 h, the volume of distribution at 20.4 +/- 6.3 l and the peritoneal clearance at 0.19 +/- 0.04 ml/min. After iterative intraperitoneal administration, serum and dialysate concentrations of piperacillin were above the minimum inhibitory concentration for susceptible pathogens without antibiotic accumulation. Peritoneal absorption was higher during peritonitis (83.4 +/- 4.8%) than without peritonitis (67.8 +/- 8.5%). Piperacillin 2 g IV every 8 hours or 1 g IP every 6 hours seemed to be the appropriate regimen in patients with chronic renal failure on CAPD. PMID- 3043347 TI - [Evaluation of the biliary excretion of a ticarcillin-clavulanic acid combination in man]. AB - The association of a beta-lactamase inhibitor, clavulanic acid (CA) (0.2 g) to ticarcillin (TIC) (3 g) enhances the activity of the latter on resistant strains. The aim of the present study was to assess their biliary elimination in man. Serum, urine and bile concentrations of TIC and CA were measured in biological samples collected in 10 cholecystectomized patients provided with a T-tube, during 12 hours after the IV administration of 3.2 g of claventin. Concerning TIC, a mean biliary peak of 177 +/- 49 (SEM) micrograms/ml was reached during the 2nd hour; the total biliary output (0-12 h) (AB) was 8.8 +/- 2.6 mg (0.28% of the administered dose), the hepato-biliary clearance CL HB 0.34 ml/min and the biological half-life, TB 1/2 1.2 h. The mean biliary peak of CA was 2.7 +/- 0.5 micrograms/ml and occurred during the first hour. AB amounted to 98.5 +/- 34.7 micrograms (0.04% of dose), CLHB to 0.10 ml/min and TB 1/2 1.2 h. In per operatively sampled serum, choledochal bile, gallbladder bile and gall-bladder wall, the following concentrations were measured 1 hour after the IV administration of 3.2 g of Claventin. TIC: 105 +/- 11; 386 +/- 66; 72 +/- 20 micrograms/ml and 36 +/- 11 micrograms/g. CA: 3.6 +/- 0.7; 5.9 +/- 1.5; 0.3 +/- 0.3 micrograms/ml and 0.1 +/- 0.1 micrograms/g. The biliary pharmacokinetic profiles allow to favorably consider the prophylactic use of Claventin in the surgery of the biliary tract as well as its therapeutic administration in biliary tract infections. PMID- 3043349 TI - [Do imipenem and cilastatin interfere with the determination of serum creatinine?]. AB - Some cephalosporin antibiotics increase the concentration of serum creatinine. This phenomenon could be due to the interference on the method of dosage, in particularly when the Jaffe reaction is used. This interaction could be present with imipenem, an antibiotic of thienamycins group having a chemical structure similar to that of cephalosporins, (associated with cilastatin in the tienam), in which we find the structure common to creatinine (Formula: see text) considered to be responsible of this interference. Our work indicates that with serum concentration of 100 mg/l, superior to those obtained in therapeutic, neither imipenem nor cilastatin interact on the creatinine dosage using either the Jaffe reaction of the enzymatic technic. PMID- 3043350 TI - [Pharmacokinetics of cefatrizine administered in repeated doses]. AB - Twelve healthy volunteers received cefatrizine orally at doses equal to 500 mg every 12 h for 5 days. Cefatrizine was assayed by high performance liquid chromatography in plasma and urines collected after the first and/or the last administration. Cefatrizine absorption was rapid; its peak plasma level was reached at time 1.79 +/- 0.07 h following the first dose, it was equal to 7.37 +/ 0.31 micrograms.ml-1. Its apparent elimination half-life was equal to 1.50 +/- 0.05 h, it explains the lack of accumulation with time during multiple administrations every 12 hours. Comparisons between peak plasma concentration and area under curves following the first and last dosing showed significant (p less than 0.01) but weak (close to 15%) reduction of these 2 parameters with time which could be explained by a slight reduction of cefatrizine absorption with time. In conclusion, cefatrizine does not accumulate when administered repeatedly at a dose equal to 500 mg every 12 h in young adult, and its pharmacokinetics is virtually linear with time. PMID- 3043351 TI - [Use of imipenem-cilastatin in neonatal septicemias caused by gram-negative bacilli multiresistant to beta-lactam antibiotics]. AB - Seven neonates with septicemia due to Gram negative bacteria resistant to beta lactam received imipenem-cilastatin therapy. Bacteria isolated were Enterobacter cloacae [3], Enterobacter aerogenes [1], Klebsiella pneumoniae [1], Serratia marcescens [1], Pseudomonas fluorescens [1]. The MICs of imipenem were lower 1 microgram/ml. In 3 children septicemia occurred during previous antimicrobial chemotherapy. 3 IV 60 mg/kg doses of imipenem with amikacin (15 mg kg/d) were administered every day. For five children blood cultures were negative after 48 hours of treatment. E. aerogenes septicemia required pefloxacin because blood cultures remained positive (d5) despite an increased dosage (90 mg/kg/d). All children were cured and imipenem-cilastatin was not responsible for any complication. Those results demonstrate the efficacy of imipenem in the treatment of septicemia in newborns due to multiresistant Gram negative bacteria. PMID- 3043352 TI - [Aztreonam treatment of severe infections caused by gram-negative aerobic bacilli]. AB - Twenty nine patients of an intensive care unit (9 women and 20 men), aged 63.9 +/ 15.8 years, with a mean body weight of 62.5 +/- 11.8 kg were treated during 9.4 +/- 2.1 days by aztreonam (2 x 1 g/24 h) administered by short infusion (30 min) for a severe infection due to a Gram-negative bacilli. The primary (n = 25) or nosocomial (n = 4) infection sites were a peritonitis (14), a septicaemia (6), a cholecystitis (6), a pyelonephritis (5), a cholangitis (2), a subphrenic abscess (1) or a pneumonia (2). The isolated Gram-negative bacilli were all susceptible to aztreonam, their MIC being less than or equal to 0.5 micrograms/ml, except for a Pseudomonas aeruginosa (MIC = 4 micrograms/ml). Aztreonam was administered as a single therapy to 7 patients and in association with metronidazole (18) and/or penicillin G (14) to 22 patients; in fact, anaerobes were isolated in ten patients. The mean serum concentrations of aztreonam, as measured by HPLC, before and after the 7th administration respectively were 83.2 +/- 17.5 and 6.1 +/- 5.5 micrograms/ml for peak and through levels. The treatment of the 29 infections was a success in all the cases. No complication occurred due to the presence of Gram positive cocci (n = 4) in the first bacteriological sample, or due to the emergence (n = 12) of Gram positive cocci, except for one case of sepsis of the abdominal wall by Staphylococcus aureus. Aztreonam (2 x 1 g/24 h) may be a suitable alternative for the treatment of severe infections of intensive care units, mostly due to Gram-negative bacilli. PMID- 3043353 TI - [Experimental Enterobacter cloacae endocarditis treated with gentamicin. Predictive value of the in vitro bactericidal rate]. AB - The predictive value of in vitro time-kill curve was tested on an Enterobacter cloacae endocarditis experimental model. The antibiotic studied was gentamicin. Despite a similar MIC, 2 Enterobacter cloacae strains exhibited very different time-kill curves in vitro. This difference was found being predictive of efficacy on the in vivo model, 24 hours after a single injection of gentamicin. PMID- 3043354 TI - [Multicenter study of the clinical efficacy and tolerance of roxithromycin compared to erythromycin ethylsuccinate in lower respiratory tract infections]. AB - In a double blind, randomised investigation in 193 hospitalized patients, with low respiratory infections, roxithromycin (150 mg bd) and erythromycin ethylsuccinate (1 g bd) were compared. Assessment of safety was made in 183 patients and clinical response in 155 patients. The mean duration of treatment was 11 days in both groups. Clinical effectiveness was 82% (67/82) for roxithromycin and 77% (56/73) for erythromycin ethylsuccinate. Roxithromycin appears to have a good effectiveness and to be effective as erythromycin ethylsuccinate. The safety profile is satisfactory in both groups. PMID- 3043356 TI - [Treatment of peritonitis in continuous ambulatory peritoneal dialysis with intraperitoneal ceftriaxone]. AB - In 16 patients under CAPD, 29 cases of bacterial peritonitis were observed, with clinical manifestations in 23. The mean cell count in peritoneal dialysis fluid was 5608/mm3 with 4991/mm3 polymorphonuclear, Leukocytes Causative pathogens were Staphylococcus in 14 cases, Streptococcus in 6, Bacillus in one, Enterobacteria in 5, Pseudomonas aeruginosa in 1 and Moraxella in 1. Three cultures were negative. First choice treatment was a daily intraperitoneal injection of 1 g of ceftriaxone. 79.3% of patients recovered within 5 days. Failure were due to a Methicillin-resistant Staphylococcus epidermidis in one case, a Streptococcus faecalis in two cases, and a Staphylococcus aureus in three observations, which two were responsible of abscess round catheter peritoneal. Mean ceftriaxone concentrations 24 hours after the intraperitoneal injection were 50.6 mg/l (range: 3.3-141 mg/l) in serum and 58.1 mg/l (range: 4.3-180 mg/l) in dialysate. These concentrations are greater than most of ceftriaxone's MICs for susceptible bacteria, a finding that confirm the value of treatment with a single daily intraperitoneal injection of ceftriaxone. PMID- 3043355 TI - [Multicenter study of the clinical efficacy and tolerance of roxithromicin compared to doxycycline in lower respiratory infections]. AB - In a double-blind, randomised investigation in 305 in-patients, roxithromycin and doxycycline were compared in two groups of patients with low respiratory tract infections. Three hundred cases were analyzable for safety and 276 for clinical efficacy. Mean duration of treatment was 9 days in both groups. Clinical effectiveness was 83% (112/135) for roxithromycin (150 mg bd) and 84% (118/141) for doxycycline (200 mg once daily), the difference not being statistically significant. A 90% clinical response rate was obtained with roxithromycin in pneumoniae. Safety and tolerance were good and comparable in both groups. PMID- 3043358 TI - [Use of ceftazidime combined with netilmicin in the treatment of febrile episodes occurring after bone marrow transplantation in children]. AB - Thirteen episodes of fever in bone marrow transplantation recipients (23 months to 11 years old children) were treated by ceftazidime (100-200 mg/kg/j) and netilmicin (7 mg/kg/j). Vancomycin was added at the 24th hour in 10 cases of persistent fever. 6 presumed agents of infection were isolated before antibiotic treatment: blood cultures (streptococci 2, staphylococcus 1, proteus 1), fecal sample (E. coli 1), urine (E. coli 1). Modifications of aerobic fecal flora were studied under this treatment. E. coli, staphylococci and enterococci were the mainly strains isolated. There were no third generation cephalosporins resistant Gram-negative bacteria. High level resistance to aminoglycosides was observed in enterococcal strains, isolated during and after treatment. Ceftazidime-netilmicin (+/- vancomycin) was an effective and safe combination for the management of febrile neutropenic episodes. PMID- 3043357 TI - [Our experience with ofloxacin in the treatment of osteoarticular infections]. AB - Twenty patients with osteoarticular infections, fourteen post-arthroplasty and six with osteitis, were treated with ofloxacin, usually in combination. Sixteen staphylococcus strains including eight aureus and eight coagulase negative (modal MIC and MBC: 0.5 micrograms/ml), three Escherichia coli (modal MIC and MBC: 0.06 micrograms/ml) and one Peptococcus (MIC: 0.25, MBC: 0.5) were isolated. Treatment was given at a mean dose of 9.81 +/- 2.46 mg/kg for a mean duration of 100 days. The serum concentration of ofloxacin was measured at 3.73 +/- 2.13 micrograms/ml for a dosage of 8.23 +/- 0.94 mg/kg (25 assays) and 7.42 +/- 4 micrograms/ml for 11.46 +/- 1.3 mg/kg (23 assays). Bacteriological control was carried out nineteen times; in one case of staphylococcal osteitis, a relapse occurred on the 43rd day of treatment when the strain isolated was resistant to ofloxacin. Three patients presented adverse effects: two cases of bone and muscle pain and one cutaneous allergic reaction: treatment was withdrawn after two restarts. The antibacterial action, the good tolerance and the easy administration of ofloxacin make it a useful antibiotic in the treatment of osteoarticular infections, where a dosage of 8 mg/kg appears to be necessary, particularly in infections due to staphylococci. PMID- 3043359 TI - [Randomized trial of empirical antibiotic therapy in febrile episodes after bone marrow transplantation. Comparison of an aminoglycoside-beta lactam (tobramycin ticarcillin) combination with 2 beta-lactam antibiotics (ticarcillin-latamoxef)]. AB - From February 1986 to July 1987, 87 patients who underwent an autologous or allogeneic bone marrow transplantation were randomized to receive ticarpen tobramycin or ticarpen moxalactam for their first febrile episode. Forty received ticarpen tobramycin and 47 ticarpen moxalactam. The two groups were similar according to age, sex, conditioning regimen, underlaying pathology and duration of granulocytopenia. We observed 58.6% of fever unknown origin and 37% of bacteremia. A similar number of clinical and bacteriological successes occurred in the two groups. Hepatic and renal toxicities were similar in the two groups. Vancomycin was widely used in these patients without particular justification. We conclude that the use of two beta lactam is a possible antibiotherapy in marrow transplantation where renal function should be preserved from additional toxicity. PMID- 3043360 TI - [Treatment with a piperacillin and netilmicin combination of patients with agranulocytosis]. AB - Sixty infections episodes in granulocytopenic patients have been treated in first line with a piperacillin and netilmicin combination. Treatment has been successful in 78% bacterial infections. So, this antibiotic combination appears as a very effective therapy of infection in neutropenic patients. PMID- 3043362 TI - [Use of a rinsing solution for contact lenses in the prevention of amebic keratitis]. AB - Recent reports incriminating Acanthamoeba, a small free-living amoeba, wide spread in environmental soils and waters, in acanthamoebic keratitis cases wearing soft contact lenses, drew attention to cleaning solutions for contact lenses. The purpose of this report is to discuss the amoebicidal action of a rinsing solution containing 0.001% thimerosal. A. castellanii cysts and trophozoites were incubated with the rinsing solution. Amoeba saline was used as control solution. After 1 h to 96 h of contact time, the survival was studied by growing of trophozoites and excystment of cysts on cultures. The results indicate that trophozoites were destroyed by the rinsing solution after 6 hours, whereas for cysts a negative culture was obtained after 72 h to 96 h. The amoebicidal and cysticidal action of this solution containing mercury corroborates our preceding experience on the effects of mercury derivates on amoebae. The use of amoebicidal solutions in contactology could inhibit and even destroy amoebae on contaminated lenses preventing severe keratitis. PMID- 3043361 TI - [Comparative action of 8 azole derivatives against Candida albicans: fungistatic action and cytologic study by scanning electron microscopy]. AB - The authors compared the in vitro antifungal activity of eight imidazole derivatives (clotrimazole, econazole, isoconazole, ketoconazole, miconazole, oxiconazole, terconazole, tioconazole) against 42 strains of Candida albicans by the agar dilution method using casitone medium. The geometric (G) mean MIC values, the MIC 90 and the MIC 50 values and the corresponding standard deviations of each antifungal agent were determined. The G-MIC values were found to be in the range of 0.008-0.390 micrograms ml-1. The effects of these eight antifungal agents on the ultrastructure of C. albicans yeast cells and spheroplasts were studied by scanning electron microscopy (SEM). The results showed a good correlation between the lesions observed and the structure of the imidazole derivatives tested. On the basis of the SEM results, the compounds could be divided into three groups: (1) ketoconazole and terconazole; (2) econazole, isoconazole, miconazole, oxiconazole and tioconazole; (3) clotrimazole. PMID- 3043363 TI - [Comparative study of 3 methods of evaluation of antiseptic products and disinfectants in quality control]. AB - For choosing a protocol of determination of bactericidal activity of antiseptic and disinfectant products, we have compared a method using an MS2 Abbott system and the classic methods: membrane filtration and dilution-method by transfer loops. The bactericidal-activity of 12 antiseptic solutions are determined. The results shown that there was no significative difference between the automated system and AFNOR specifications. The dilution-method by transfer loops gave higher bactericidal concentrations than the two other methods. PMID- 3043364 TI - [Health care in the schools. I and II]. PMID- 3043365 TI - Immunosuppressive effects and infections associated with corticosteroid therapy. PMID- 3043366 TI - Bartholin's gland abscess in infancy. PMID- 3043367 TI - Primary immunization of infants with an acellular pertussis vaccine in a double blind randomized clinical trial. AB - The rate of adverse reactions and the immunogenicity of a two-component acellular pertussis vaccine as compared with a plain whole-cell vaccine and a placebo were evaluated for primary immunization in 319 6-month-old infants in a double-blind randomized clinical trial. The acellular vaccine produced few and mild systemic and local reactions. Fever (greater than or equal to 38 degrees C) occurred in 6% to 8% of acellular vaccinees as opposed to 25% to 37% of whole-cell vaccinees. Redness (greater than or equal to 1 cm) appeared in 2% to 13% of the acellular vaccine and 24% to 32% of the whole-cell vaccine recipients. Antibody response to pertussis toxin measured in a neutralization test was obtained in 97% to 100% of the infants receiving either two or three doses of the acellular vaccine as compared to 59% after three doses of whole-cell vaccine. PMID- 3043369 TI - Preventive care use by school-aged children: differences by socioeconomic status. AB - Use of ambulatory care services by children from low-income families has increased substantially since the early 1960s. However, in few studies have attempts been made to disaggregate physician visits according to type (eg, preventive upsilon diagnosis and treatment). In this study, receipt of preventive care (including physical, vision, and dental examinations), based on a sample of 16,838 children aged 5 to 16 years from the 1982 National Health Interview Survey, was examined. The results indicate that children in families with incomes below the poverty level, especially those without Medicaid insurance, are much less likely to receive routine preventive care on a timely basis. Poor school aged children with Medicaid are much more likely to receive timely preventive care than their counterparts without Medicaid coverage. The effectiveness of preventive care for children is discussed and suggestions for improving access to routine preventive care are presented. PMID- 3043368 TI - Predictive value of early continuous electroencephalogram monitoring in ventilated preterm infants with intraventricular hemorrhage. AB - The contribution of early continuous four-channel EEG monitoring to the evaluation of intraventricular hemorrhage in acutely ill preterm infants mechanically ventilated for acute respiratory distress was assessed in a prospective study of 54 infants of less than 34 weeks' gestation. Early abnormal EEG results correlated significantly with later outcome. They often preceded ultrasound evidence of hemorrhage and provided prognostically significant functional correlation with the grade of hemorrhage. Continuous EEG monitoring allows collection of significant data with minimal interference and could contribute to clinical management of high-risk preterm infants. PMID- 3043370 TI - Motor performance in hyperactive children treated with imipramine. AB - The effects of the tricyclic antidepressant imipramine were evaluated in a study of 9 children with Attention Deficit-Hyperactivity Disorder. The study was double blind, placebo-controlled, with three drug conditions, low, medium, and high doses. The focus was on neuropsychological drug effects. Imipramine exerted negative dose-response effects on motor performance (motor speed, motor pursuit), while it improved hyperactive behavior and attention and raised the heart rate slightly. PMID- 3043371 TI - Effects of relaxation response training on attentional deficits in schizophrenics. AB - Four groups of six male patients each (12 = schizophrenic; 12 = antisocial personality) volunteered. One group from each diagnostic category were instructed in the Relaxation Response while the other two groups were instructed with a placebo exercise. After training, subjects were exposed to an attentional task involving manual responding to visual stimuli. Analyses of several dependent measures including mean correct responses yielded no significant group differences. PMID- 3043372 TI - Contributions to the history of psychology: XLVIII. Ancient Greek roots of the assumptions of modern clinical psychology. AB - This paper is an account of studies of the linguistic transformation that took place in ancient Greece between the eighth and fourth centuries B.C., searching for factors which contribute to the shift in how humans perceived themselves. The group or force-field consciousness of the men of the Iliad and the linguistic factors which allowed "individuality" to emerge by the time of Plato is explored. The account relates the emergence of the notion of "madness" to the development of the individual and asks whether madness is an artifact of individuality and explores the relationship of these developments to our present underlying assumption of a duality in human nature composed of the rational and the irrational. PMID- 3043373 TI - Moral reasoning and ethical practice in nursing: an integrative review. PMID- 3043374 TI - Sequence-directed bends of DNA helix axis at the upstream activation sites of alpha-cell-specific genes in yeast. AB - The MF alpha 1 gene of Saccharomyces cerevisiae, a major structural gene for mating pheromone alpha factor, is an alpha-cell-specific gene whose expression is regulated by the mating-type locus, MAT. Two upstream activation sites (UASMF alpha 1s), which are binding sites for an activator protein, MAT alpha 1, mediate alpha-cell-specific expression of this gene. We show here that DNA fragments containing the UASMF alpha 1 region exhibited anomalous slow electrophoretic mobilities on gels at lower temperature, but not at higher temperature, that is characteristic of bent DNA. We confirmed the sequence-directed bend at the UASMF alpha 1 region by employing a circular permutation analysis and a DNA cyclization assay. Deletion analyses revealed the existence of two bends in this region, each of which overlaps each UASMF alpha 1 element. The two bends were almost in phase; they lie in a nearly same plane with a same direction. We also show the existence of sequence-directed bend at the UAS region of the STE3 gene, another alpha specific gene in S. cerevisiae. These bent DNAs may be involved in transcriptional regulation of this set of genes. PMID- 3043376 TI - Genetic hypervariability of telomere-related sequences is associated with meiosis in Plasmodium falciparum. AB - Sequences related to those near chromosome telomeres in the human malaria parasite, Plasmodium falciparum, were extremely unstable during a genetic cross between two different clonal genotypes. Many progeny of the heterologous cross displayed telomere-homologous restriction fragments found in neither parent. A significant number of the new fragments resulted from rearrangements at chromosome-internal locations which were bounded by more complex tracts of DNA sequence. The same instability was not seen to arise during an inbreeding cross, nor during mitotic replication of parasites. Thus, a form of genetic hypervariability results from molecular events which occur during meiotic reduction and is apparent only in a cross between heterologous strains of parasite. Since other sequences were entirely stable under the same conditions, it appears that chromosome-internal blocks of telomeric sequences in the P. falciparum genome may designate conditionally unstable chromosomal domains. We discuss some potential implications of these findings for the population biology of P. falciparum. PMID- 3043375 TI - Identification of a protein factor binding to the 5'-flanking region of a tRNA gene and being involved in modulation of tRNA gene transcription in vivo in Saccharomyces cerevisiae. AB - Control mechanisms of tRNA gene transcription were studied in vivo in Saccharomyces cerevisiae. In order to be able to monitor in vivo transcription products of an individual tRNA gene, a 'tester gene' was used which is readily transcribed in vivo in yeast but does not cross-hybridize with any cellular yeast tRNA. A series of insertion mutants were constructed, modifying thereby the immediate and further distant 5'-flanking region of the 'tester tRNA gene'. Small linker molecules of different length and different sequence were inserted at positions -3 and -56 on the non-coding strand. Resulting tRNA gene variants were transformed into yeast cells and in vivo synthesized products were monitored by primer extension analysis. From the experimental data we suggest that a few essential nucleotides within the flanking region are able to determine the in vivo transcription activity of the 'tester tRNA gene'. Our results are rationalized on a biochemical level by protein binding assays: At least one protein binds to the 5'-flanking region of the 'tester tRNA gene' and different protein complexes are sequestered on active or less active tRNA gene variants. PMID- 3043377 TI - Reconstruction of the yeast 2 micron plasmid partitioning mechanism. AB - The effect of the yeast 2 micron circle encoded REP1 and REP2 gene products on plasmid partitioning and copy number control was analyzed by removing the open reading frames from their normal sequence context and transcriptional control regions and directing their expression using heterologous promoters in [cir0] host strains. Both the REP1 and REP2 gene products are directly required at appropriate levels of expression to reconstitute the 2 microns circle partitioning system in conjunction with REP3 and the origin of replication. The level of expression of REP2 appears to be critical to re-establishing proper partitioning and may also play a role in monitoring and thereby regulating the plasmid copy number. Constitutive expression of the REP1 and REP2 open reading frames using heterologous expression signals can be used to reconstruct efficient plasmid partitioning even in the absence of FLP-mediated plasmid amplification and a functional D open reading frame. PMID- 3043378 TI - Mono- through hexanucleotide composition of the sense strand of yeast DNA: a Markov chain analysis. AB - Here we compare several methods for predicting oligonucleotide frequencies in 392 kb of yeast DNA. As in previous work on E. coli, a relatively simple equation based on tetranucleotide frequencies can be used in predicting the frequencies of longer oligonucleotides. For example, the mean of observed/expected abundances of 4,096 hexamers was 1.00 with a sample standard deviation of .18. This simple predictor arises by considering each base on the sense strand of yeast to depend only on the three bases 5' to it (a 3rd order Markov chain) and is more accurate in estimating oligonucleotide frequencies than other statistical methods examined. This equation is useful in predicting restriction enzyme fragment sizes, selecting restriction enzymes that cut preferentially in coding vs noncoding regions, and in constructing detailed physical maps of whole genomes. When ranked highest to lowest abundance, the observed frequencies of oligomers of a given length (up to 6 bases) are closely tracked by the predicted abundances of a 3rd or 4th order Markov chain. These ordered abundance curves have a power curve shape with a broad linear range with a sharp break at the top end of the curve. There is also a strong disparity between the most and least abundant oligomer with for example a 79-fold variation between the most and least abundant hexamer. The curves reveal a strong dependence of oligomer frequencies on base composition. Unlike E. Coli, there is no sharp downturn at the low end of the curves and hence, no class of oligomers rare relative to other oligomers of the same length. PMID- 3043379 TI - The N49-N65 base-pair could play an important role in adaptation between tRNAs and aminoacyl-tRNA synthetases. PMID- 3043380 TI - The primary structure of human adenovirus type 12 protease. PMID- 3043381 TI - Nucleotide sequence of the HU-1 gene of Salmonella typhimurium. PMID- 3043382 TI - Nucleotide and deduced amino acid sequences of the Klebsiella pneumoniae nifK gene coding for the beta-subunit of nitrogenase MoFe protein. PMID- 3043383 TI - Capillary filtration in idiopathic cyclic edema--effects of Daflon 500 mg. PMID- 3043384 TI - 40 years of the NHS. Life from the inside. Interview by Jane Feinmann. PMID- 3043385 TI - 40 years of the NHS. We don't do that sort of thing here. PMID- 3043386 TI - RCM Supplement. Sound judgement. PMID- 3043387 TI - Calcium and colon cancer: a review. AB - The role of dietary calcium as a protective factor in the etiology of colon cancer is reviewed by examining data from ecological and analytical epidemiological studies. Biological evidence that explains the mechanisms whereby calcium intake could alter risk of developing colon cancer is also presented. The data reviewed here in general support the hypothesis that dietary calcium is linked to colon cancer in a protective manner, and that it may be one component in the etiology of colon cancer which alters an individual's risk of developing the disease. PMID- 3043388 TI - [Identification of the causative agent of allergic alveolitis in a selected group of patients by using microbiological and immunological methods]. PMID- 3043389 TI - [Allergy to insect venoms]. PMID- 3043390 TI - Mechanism of hypoxic pulmonary hypertension. PMID- 3043391 TI - [Lysosomal enzymes and their natural inhibitors in the serum and bronchoalveolar lavage fluid in chronic bronchitis and pneumonia]. PMID- 3043392 TI - [The physiological role of L-carnitine in the human body: causes and effects of its deficiency]. PMID- 3043393 TI - [Pituitary-testicular axis in patients with transplanted kidney treated with cyclosporin with prednisone or azathioprine with prednisone]. PMID- 3043395 TI - [Cellular engineering--a new trend in clinical medicine]. PMID- 3043397 TI - [Medical genetic engineering or molecular biology used in the diagnosis and treatment of disease]. PMID- 3043398 TI - [Adoptive immunotherapy of neoplasms]. PMID- 3043396 TI - [Allogeneic transplantation of HLA-compatible bone marrow. Initial results]. PMID- 3043394 TI - [Endourology: progress in the treatment of calculi of the upper urinary tract]. PMID- 3043399 TI - [Transplantation of epidermis grown in vitro]. PMID- 3043400 TI - [Clinical evaluation of fendiline as a coronary drug]. PMID- 3043401 TI - [Pharmacological treatment of primary degenerative dementia]. PMID- 3043402 TI - [Bleeding time: critical review of methods and interpretation]. PMID- 3043403 TI - [Biology and clinical aspects of primary myelodysplastic syndromes]. PMID- 3043404 TI - [Is arteriosclerosis reversible]. PMID- 3043405 TI - [Effect of the treatment with captopril and verapamil on left-ventricular function in patients with primary arterial hypertension]. PMID- 3043406 TI - [Clinical usefulness of the analysis of beta 2 microglobulin]. PMID- 3043407 TI - Three dimensional imaging in orthopedics: state of the art 1988. PMID- 3043408 TI - Simultaneous dorsal dislocation of the interphalangeal joints in a finger. Case report and review of the literature. AB - A case report and review of the English literature on the simultaneous dorsal dislocation of the interphalangeal joints of the finger are presented. This injury usually occurs on the ulnar side of the hand of young male athletes. It is easily reduced by a closed technique, although slight limitation of motion and joint swelling may persist. PMID- 3043409 TI - Effects of kanamycin administration to poultry on the proliferation of drug resistant Salmonella. AB - Four experiments were conducted to examine the relationship between antibiotic administration to poultry and the in vivo proliferation of Salmonellae. The frequency of isolation of drug-resistant transconjugant S. arizonae from the livers of chicks inoculated per os with multiply drug-resistant Escherichia coli and drug-sensitive S. arizonae was directly related to the concentration of kanamycin administered to the chicks in their drinking water. Kanamycin administration was also associated with a significant (P less than .05) increase in the frequency of isolation of drug-resistant transconjugant S. typhimurium from the intestines and livers of poults inoculated with drug-sensitive S. typhimurium and multiply drug-resistant E. coli. Kanamycin administration significantly reduced the spread of drug-sensitive S. typhimurium to the livers of poults inoculated only with that strain. These experiments demonstrate that antibiotic administration to poultry can enhance the proliferation of drug resistant Salmonella. PMID- 3043410 TI - Effects of kanamycin administration to poultry on the interspecies transmission of drug-resistant Salmonella. AB - Three experiments were conducted to assess the relationship between antibiotic administration to poultry and the transmission of drug-resistant Salmonella through a simulated food chain. Poults were inoculated per os with either multiply drug-resistant Escherichia coli and drug-sensitive S. typhimurium or with multiply drug-resistant S. typhimurium. One-half of the poults inoculated with drug-sensitive S. typhimurium and all poults given drug-resistant S. typhimurium received kanamycin in their drinking water. Liver tissue from these poults was incorporated into diets fed to rats, half of which were treated with kanamycin. Antibiotic administration to the poults was associated with a significantly (P less than .05) higher frequency of transmission of drug resistant S. typhimurium to the rats through the simulated food chain. S. typhimurium was isolated only from rats treated with kanamycin. The highest frequency of isolation of drug-resistant S. typhimurium from rats (40%) was observed in kanamycin-treated rats fed a diet containing liver from kanamycin treated poults. Results of this experiment indicate that antibiotic administration can increase the frequency of transmission of drug-resistant Salmonella through the food chain. PMID- 3043411 TI - [Corticoid-induced osteoporosis]. PMID- 3043412 TI - [Pleural mesothelioma with coincidental second cancer. Documentation of 7 cases and review of the literature]. PMID- 3043413 TI - [Deafness from the viewpoint of the child and adolescent psychiatrist. 1: Medical aspects, epidemiology and development psychology aspects]. PMID- 3043414 TI - First trimester chorionic villus sampling versus mid-trimester genetic amniocentesis--preliminary results of a controlled prospective trial. AB - This controlled prospective study assesses the relative risks of first trimester chorionic villus sampling (CVS) versus mid-trimester gentic amniocentesis (GA). CVS subjects and amniocentesis controls were comparable with regard to several confounding variables which might influence the risk of pregnancy loss including maternal age, smoking, alcohol consumption, gestational age at study entry, and history of vaginal bleeding or poor prior reproductive outcome. The most common indication for prenatal diagnosis was advanced maternal age (n = 511). In this subgroup, spontaneous abortion (less than 24 weeks) occurred in 2.9 per cent of CVS subjects versus 4.3 per cent of amniocentesis controls. The sum of spontaneous and therapeutic abortions (less than 24 weeks) was identical (5.3 per cent) in both groups. Therefore, intervention in the CVS group (i.e., therapeutic abortion for cytogenetic abnormalities) did not influence the observed risk of pregnancy loss. Overall perinatal mortality rates were also similar in both groups. No significant differences were identified for a number of pregnancy outcome parameters including 5 min Apgar score, birth weight, body length, head circumference, gestational age at delivery, preterm delivery, fetal growth retardation, congenital malformations, and neonatal complications. Preliminary results of this controlled prospective study suggest that chorionic villus sampling carries a low and acceptable risk. PMID- 3043415 TI - Prenatal diagnosis of cystic fibrosis: ultrasonographic appearance of meconium ileus in the fetus. AB - Four of ten fetuses carrying a risk of 1:4 for cystic fibrosis were found to have low levels of microvillar enzymes in the amniotic fluid obtained between 17 and 18 weeks' gestational age. On sonography performed prior to the amniocentesis, three fetuses showed enlarged bowel loops. At autopsy, meconium ileus was detected. Enlarged bowel loops are a sign which has not been described previously so early in pregnancies. PMID- 3043416 TI - [Critical moments in the formal pathogenesis of the rejection reaction in the orthotopic heart transplant with prognostic significance]. PMID- 3043417 TI - [Schiller's disease and death from the pathologico-anatomic and clinical viewpoints]. PMID- 3043419 TI - Sporozoite immunity and vaccine development. PMID- 3043418 TI - Mortality patterns among hypertensives by reported level of caffeine consumption. AB - The effect of caffeine consumption on mortality was evaluated in a historical cohort study of 10,064 diagnosed hypertensive individuals participating in the Hypertension Detection and Follow-up Program from 1973 to 1979. Total caffeine intake level from beverages (coffee and tea) and certain medications, was estimated at the 1-year visit. No evidence was found supporting an association between increased level of caffeine consumption and increased all-cause mortality or cardiovascular disease mortality during the following 4 years. Cigarette smoking was significantly associated with mortality; the association being more pronounced among non- and low-caffeine consumers for all-cause mortality and among non-caffeine consumers for all cardiovascular mortality except cerebrovascular mortality. PMID- 3043420 TI - Biology and immunology of sporozoite invasion of liver cells and exoerythrocytic development of malaria parasites. PMID- 3043421 TI - T cell functions in Plasmodium falciparum and other malarias. PMID- 3043422 TI - Interactions between chemotherapy and immunity to malaria. PMID- 3043423 TI - Invasion of erythrocytes by malaria parasites: erythrocyte ligands and parasite receptors. PMID- 3043424 TI - The precursor to major merozoite surface antigens: structure and role in immunity. PMID- 3043425 TI - Malarial proteins at the membrane of Plasmodium falciparum-infected erythrocytes and their involvement in cytoadherence to endothelial cells. PMID- 3043426 TI - Platelet-activating factor in graft rejection. PMID- 3043427 TI - Ether lipids in the therapy of cancer. PMID- 3043428 TI - PAF-acether and eosinophils. PMID- 3043429 TI - Role of PAF-acether in the vascular events of inflammation. PMID- 3043430 TI - Proinflammatory activity of platelet-activating factor: pharmacological modulation and cellular involvement. PMID- 3043431 TI - PAF-acether and kidney pathology. PMID- 3043432 TI - Platelet-activating factor and shock. AB - The aim of this chapter was to highlight the major components of PAF actions which lead to a state of shock, i.e. inadequate perfusion of essential organs which if sustained over a critical period of time, leads to irreversible damage in essential organs and eventually death. The heart, the pulmonary vessels and the microcirculation seem to be the primary target organs to PAF-induced hypotension. The effects of PAF on the pulmonary airways in some species (bronchoconstriction) might lead to hypoxemia and further exacerbate organ function. Thrombocytopenia, leukopenia and activation of the complement system are also important in PAF-induced shock by promoting thrombi formation and generation of multiple secondary mediators (e.g. histamine kinins, TXA2, leukotrienes, oxygen radicals). Identification of PAF production during specific or generalized pathophysiological processes is a critical step to implicate this vasoactive lipid in disease processes. So far, only limited information has been derived from studies involving immune responses (anaphylaxis) or bacterial endotoxins. Yet, the growing number of selective and potent PAF antagonists provide important information on the potential role of PAF in shock states. Such evidence, summarized in table I, is of great importance in designing new therapeutic strategies to a highly complex and lethal disease such as septicemia. However, the data summarized in table I clearly show that little is known on the mechanism of action of the various PAF antagonists. It is also important to note that PAF-induced shock and death can be prevented by drugs which are not necessarily PAF antagonists. For example, dexamethasone is extremely efficient in preventing PAF-induced shock and death in the mouse [24, 39] and thyrotropin releasing hormone in the guinea pig [15]. Therefore, it is conceivable that pathological conditions in which PAF might play a fundamental role might be reversed by pharmacological interventions which activate physiological mechanisms which can overcome and reverse the pathological processes activated by PAF. In conclusion, PAF is a powerful vasoactive lipid which can produce severe derangements in essential biological functions which can lead to death. The role of PAF in pathological processes in vivo is well supported in conditions such as anaphylaxis and endotoxemia. Yet, direct proof for PAF production in other shock states, such as multiple trauma, ischemia, inflammation and hemorrhage, is still missing. Furthermore, it is important to keep in mind that in shock, trauma or inflammation, multiple mediators in addition to PAF are formed.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3043433 TI - Chemistry of PAF antagonists. PMID- 3043434 TI - Formation and biological effects of fecapentaenes. PMID- 3043436 TI - Pharmacology of PAF antagonists. PMID- 3043435 TI - Biological effects of alkylglycerols. PMID- 3043437 TI - PAF in cellular immunology. PMID- 3043438 TI - Effects of PAF on neutrophils and mononuclear phagocytes. PMID- 3043440 TI - Regulation of calcium uptake/efflux from the islet-cell endoplasmic reticulum with regard to the secretion of insulin. AB - Figure 8 summarizes some of the processes that may impact on the secretion of insulin by regulating Ca2+ handling by the beta-cell endoplasmic reticulum. A role for calmodulin in controlling the rate of Ca2+ efflux is indicated by both the ability of the calmodulin antagonist, W7 to stimulate Ca2+ efflux and by the ability of exogenous calmodulin to antagonize Ca2+ efflux in response to inositol trisphosphate (IP3). The impact of calmodulin on this system may be to serve as link in the feedback control of cellular Ca2+. In addition to IP3, a second messenger that may link signal transduction to the release of Ca2+ is the guanine nucleotide, GTP. GTP stimulates the efflux of Ca2+ from the endoplasmic reticulum through a mechanism distinct from IP3. It will be important to determine whether extracellular glucose concentration, or other modifiers of secretion, acutely regulate the GTP concentrations in the beta-cell and to assess if this function may be altered with a decrease in beta-cell function. A variety of evidence indicates that metabolism of glucose by the beta-cell somehow plays a major role in the cellular control of insulin secretion (Hedeskov, 1980). An important link in this process may be the direct effects of glucose 6-phosphate on the handling of Ca2+ by the endoplasmic reticulum. Glucose 6-phosphate is able to increase the active uptake of Ca2+ by these membranes and also to specifically inhibit Ca2+ efflux produced by the IP3. Concentrations of glucose 6-phosphate needed to achieve these effects are likely achieved under physiological conditions (Aschroft et al., 1970). It is also easy to imagine that in diabetes when the islets are chronically exposed to high glucose that, as a result of the content of the high Km glucokinase in the islet (Meglasson and Matschinsky, 1986), higher concentrations of glucose 6-phosphate may be achieved. Under these conditions glucose 6-phosphate may contribute to the islet-cell pathology by interfering with the acute control of Ca2+ handling by the endoplasmic reticulum. PMID- 3043439 TI - Pancreatic islet defects in NIDDM. AB - While circulating insulin levels may at times appear to be normal or even elevated in patients with NIDDM, depending on the control group for comparison and the glucose level at which subjects are studied, a profound impairment of pancreatic B-cell function is characteristic of NIDDM and contributes to the hyperglycemia in this condition (Halter, et al., 1985). Table I lists some of the defects of pancreatic islet function that have been observed in NIDDM. PMID- 3043442 TI - Critique of current therapies of noninsulin-dependent diabetes mellitus. AB - In summary, there are a number of therapeutic modalities, including life-style modifications and pharmacological intervention, available and useful in the treatment of NIDDM patients. Nonetheless, therapeutic success in the management of NIDDM remains highly dependent upon patient motivation, cooperation, and ability to comply on a regular basis with measures that are inconvenient and of modest efficacy in reversing the physiologic abnormalities of the NIDDM state. There remains considerable need for therapeutic tools of greater efficacy and of greater ease of application than the ones we currently have. PMID- 3043441 TI - Non-beta-cell islet abnormalities in noninsulin-dependent diabetes mellitus. AB - Although there are abnormalities in the function of pancreatic alpha, delta, and PP cells in NIDDM, only in the case of the glucagon-secreting alpha cells is there sufficient evidence to indicate that these abnormalities may be metabolically important. But the cause of abnormal glucagon secretion remains to be established. Studies of delta-cell secretion have been hampered by the inability to determine the source of circulating somatostatin-like immunoreactivity and the failure to distinguish between the potential molecular species being measured. Pancreatic polypeptide remains a hormone in search of a metabolic function; the main use of its measurement may be in the study of parasympathetic nervous system function in NIDDM. PMID- 3043443 TI - The hypoglycemic effect of ciglitazone in obese, hyperglycemic animal models. PMID- 3043444 TI - A review of the effects of ciglitazone on the pancreatic islets of obese, hyperglycemic mice. PMID- 3043446 TI - Possible post-receptor defects in NIDDM: second messengers. PMID- 3043445 TI - The sympatho-adrenal system and NIDDM. PMID- 3043447 TI - The role of brown adipose tissue in the development of the obese-hyperglycemic syndrome in mice. PMID- 3043448 TI - Bone marrow transplantation for poor prognosis neuroblastoma. AB - These studies suggest that intensive chemoradiotherapy (eg., VAMP-TBI), if given relatively soon after diagnosis and before development of progressive disease, improves long-term survival for patients with advanced neuroblastoma. Although this particular therapy is not useful for those who already have developed progressive disease, other intensive regimens may warrant testing in this latter group of patients. Our current study is testing aggressive induction chemotherapy, ex vivo purging of autologous marrow, and VAMP-TBI followed by BMT. Because of the increasing risk of progressive disease with time after diagnosis, we recommend beginning the BMT phase by 20 weeks after diagnosis. This should result in 85% of newly diagnosed patients entering the BMT phase without progressive disease; and, with fewer toxic deaths, 90% should survive the BMT phase. Thus, approximately 70% of patients could be disease-free survivors 8-9 months after diagnosis. Because a significant number of patients still develop progressive disease during induction or after BMT, efforts are being made to improve induction and pretransplant therapies and to identify prognostic factors. If modifications of the current regimens do not decrease the rate of progressive disease, it may be necessary to develop additional or different therapy for patients identified to be at high risk. Hopefully, new strategies will further increase the percentage of patients with tumor-free survival. PMID- 3043449 TI - Bone marrow transplantation (BMT) for advanced neuroblastoma (NBL): a multicenter POG pilot study. PMID- 3043450 TI - Very long delay to engraftment after ABMT in neuroblastoma patients and effect of CD8 monoclonal antibody in vivo therapy. PMID- 3043451 TI - In vitro treatment of autologous bone marrow for neuroblastoma patients with anti GD2 monoclonal antibody and human complement: a pilot study. PMID- 3043452 TI - Molecular analysis and clinical significance of N-myc amplification and chromosome 1p monosomy in human neuroblastomas. PMID- 3043453 TI - Phase II studies of combinations of drugs with high dose carboplatin in neuroblastoma (800 mg/m2 to 1 g 250/m2): a report from the LMCE group. PMID- 3043454 TI - Immunotherapy with GD2 specific monoclonal antibodies. PMID- 3043456 TI - The origin and evolution of atmospheric oxygen. PMID- 3043455 TI - The therapeutic use of I-131 meta-iodobenzylguanidine (mIBG) in neuroblastoma: a phase II study in 12 patients. PMID- 3043457 TI - New concepts of the molecular pathogenesis arising from hypoxia. PMID- 3043459 TI - A possible radical mechanism for the functioning of flavoprotein phenolic hydroxylases. PMID- 3043458 TI - Structural determinants of the oxygen reactivity of different classes of flavoproteins. PMID- 3043460 TI - Involvement of oxo-bridged binuclear iron centers in oxygen transport, oxygen reduction, and oxygenation. AB - The occurrence of an oxo-bridged binuclear iron site is well-established for the oxygen transport protein, hemerythrin, and strongly implicated in ribonucleotide reductase, purple acid phosphatase, ferritin, and methane monooxygenase. Key identifying characteristics are an antiferromagnetic interaction between the two iron atoms, an Fe-O-Fe vibrational mode in the resonance Raman spectrum, and an S = 1/2 EPR signal upon one-electron reduction. In hemerythrin the oxo bridge serves as a hydrogen bond acceptor which stabilizes the bound hydroperoxide. In ribonucleotide reductase both the binuclear iron center and a protein tyrosine undergo oxidation in the presence of molecular oxygen, whereas in methane monooxygenase a binuclear iron moiety may activate O2 for substrate oxygenation. PMID- 3043461 TI - An exercise in nostalgia on the theme of David Keilin. PMID- 3043462 TI - My personal encomium to Professor David Keilin. PMID- 3043464 TI - David Keilin and the Molteno Institute family. PMID- 3043463 TI - Evolution of blue copper proteins. AB - The evolutionary relationships of blue copper proteins are reviewed. Five homologous families of small blue proteins are recognized. Despite differences in length their peptide chains can all be accommodated into the eight-stranded fold of plastocyanin with some adjustments at three of the loops and the two termini. The C-termini of the blue oxidases ceruloplasmin and Neurospora laccase also fit into this fold and they are suggested to be homologous to the small blue proteins. The alignment of their amino acid sequences suggest some of the histidines to be binding active site copper. A superposition of the structures of poplar plastocyanin and bovine Cu-Zn superoxide dismutase (SOD) showed that 68 out of 99 alpha-carbons in plastocyanin overlapped with corresponding atoms in SOD with a rms distance of 2.99 A. In addition three of the histidine residues that were proposed to be copper-binding in laccase and ceruloplasmin aligned with ligands to the Cu-Zn pair in a SOD. Thus also SOD might be related to the blue proteins. PMID- 3043465 TI - 'Integrity of structure' and 'latent life'--the respiratory chain, Claude Bernard, and David Keilin. PMID- 3043466 TI - My days with David Keilin and doing science in China. PMID- 3043467 TI - The paradox of oxygen: thermodynamics versus toxicity. PMID- 3043469 TI - NMR imaging in theory and in practice. PMID- 3043468 TI - Prostaglandins and the release of insulin. A review and a proposal. AB - The effect of prostaglandins or the inhibition of their synthesis on the release of insulin is controversial. Dispute exists because there are apparently disparate experimental results. When the following factors are considered, however, much of the disparity is eliminated: 1) the experimental setting--in vitro or in vivo; 2) the experimental model--animal or human; 3) the experimental additive--the type of nonsteroidal antiinflammatory drug or specific prostaglandin; 4) the relationship of insulin levels to insulin secretion, degradation, and the observed hypoglycemic response. On the basis of such considerations the following conclusions are advanced. 1) From animal studies in vitro it appears that prostaglandins can directly augment insulin release. 2) Results from animal experiments in vivo, however, suggest that systemic prostaglandin administration diminishes insulin release. 3) No human studies have been performed in vitro which examine the insulin secretory response of pancreatic tissue to prostaglandins. 4) Prostaglandins reduce stimulated insulin levels in normal human subjects and in those with diabetes mellitus. Whether insulin secretion is reduced, or clearance is increased, is unknown. 5) Finally, the critical experiment remains to be done, that is the simultaneous examination of insulin, C-peptide, and glucose kinetics during an infusion of a prostaglandin. PMID- 3043470 TI - [Phlebology consultation services open to physicians]. PMID- 3043471 TI - [Contribution of phlebography in the surgical treatment of chronic venous insufficiency of the lower extremities]. AB - In which circumstances does a surgeon request phlebograms and which ones? What is the contribution of phlebography regarding therapeutic indications and surgical technique. PMID- 3043472 TI - [Therapeutic possibilities for the diabetic and ischemic foot]. AB - The prognosis and treatment of a diabetic foot depend on the extent of the lesions. If infected, these lesions are always mixed, secondary to aerobic and anaerobic bacteroides. The antibiotic treatment must be well adjusted and applied for a long enough time. It is absolutely necessary, for the treatment to be effective, to know the level of the ischemia in order to decide whether a lesion or an amputation scar will heal without revascularization. The author discusses various non-invasive examinations. Today, distal revascularization if possible due to new surgical techniques. But, one must emphasize that the patency of an adequate by-pass is definitely longer than the survival of a diabetic patient, in general. A accurate diagnosis and a multiudiscipline treatment will therefore improve the quality of the survival of the patient. PMID- 3043473 TI - [Medical imaging in phlebology]. PMID- 3043474 TI - [X-ray computed tomography in venous pathology]. AB - After presenting the tomodensitometric semiology of venous thromboses in various territories, the authors define its indications. It is essentially in deep territories, head, chest and abdomen that the CT-Scan is used to study and diagnose unrecognized thromboses as well as determine the venous repercussion of various neighboring ailments. The respective place of the CT-Scan, ultrasonography and various phlebographies is analyzed. PMID- 3043475 TI - [Ultrasonics and directional Doppler. Study of superficial phlebitis of the lower extremities]. AB - The objective of the study was to evaluate the usefulness of ultrasonic methods: ultrasonography and Doppler C.W. in the diagnosis of superficial varicose phlebitis. Especially, measurement of the venous pressure (V.P.) at the posterior tibial (P.T.) and the greater saphenous vein (G.S.V.) in clino and orthostatism, have proved to be useful. In ten cases, one patient with a varicose in the thigh, was also found to have a femoral vein thrombosis secondary to a deep thrombus entering the junction. In conclusion, these non-invasive methods are advisable, especially in patients presenting a varicose phlebitis at the thigh. PMID- 3043476 TI - [Ultrasonics and deep vein thrombosis. Ilio-caval level and the lower extremities]. AB - All the important abdominal veins and limbs vein can be examined. The veins, in cross-section are rounded in repletion, (maximum normal vein caliber has reached when erect) almost flat in a state of vacuous-ness. The lumen normally echofree is limited by a wall thinner and less echogenic than the wall of adjacent artery. The parietal motions are rythmed by breathing. Venous blood flow can become echogenic, with weak echoes, as snow storm within the lumen. These phenomena are often visible in venous confluent and within the lumen below occlusion where there is sludge. In supine position deep abdominal veins, limbs veins until popliteal veins are easily seen. Below the popliteal fossa in prone position the veins are nearly empty and not visible beyond pathological circumstances. Objective studies demonstrated the inaccuracy of clinical diagnosis of deep venous thrombosis, echotomography (coupled with doppler) is among the non invasive methods the most interesting, permitting to recognize venous occlusion complete or incomplete by clot but also compression by tumour or ganglion. The compression under the probe collapse the normal vein, but if there is clot inside, the compression become incomplete or impossible. The caliber of the vein is dilated also in supine position. The richest of venous clot in red cells in comparison with arterial thrombus make it more and earlier echogenic and more especially as the investigation is performed with high frequency probe. In the same way if the clot is floating its motions are put in evidence. Echotomography make usually difference between clot and neoplastic thrombus which is again more echogenic and also have special location. Echotomography permit to follow evolution ot venous thrombosis under treatment. The wall vein lesions after thrombosis are analysed showing thickening, destruction of the cups, dilatation of some veins while others are still obstructed. PMID- 3043477 TI - [Symptomatology and diagnosis of the diabetic foot]. AB - In the diabetic foot, the vascular component was recently considered as the most important in its pathogenesis. The contribution of the neuropathy was under evaluated, most of the time. However, in the last few years, the neuropathic modifications rank first and the micro- and macro-angiopathies are considered more and more as complicating components. Along with sensory and motor neuropathies, the so-called neuropathic edema is an important symptom. This edema, hot most of the time, is caused by a paralysis of the sympathetic nerve fibres, causing an increased Arterio-venous "shunt", at the expense of the capillary nutritive blood circulation. Orthostasis components and he possible presence of venous insufficiency aggravate he edema. The influence of cardiac components must be excluded. Besides, the likelihood of a venous thrombosis is not increased in diabetic patients. A multi-disciplinary approach is advocated in the diabetic foot. PMID- 3043478 TI - [Phlebography in chronic venous insufficiency of the lower extremities. Technic and value of different tests]. AB - Remainder of the various phlebographic procedures in chronic venous insufficiency of the lower extremities: peripheral phlebography, popliteal phlebography, femoral phlebography, varicography. The techniques and the informations they provide are presented for each one of these examinations. PMID- 3043479 TI - [Consultation services in phlebology open to physicians]. PMID- 3043481 TI - [Echotomography study of the sapheno-femoral venous confluence]. PMID- 3043480 TI - [Buerger's disease. Clinical and therapeutic review. Apropos of a case]. AB - In reference to a typical case of Buerger's disease, the clinical, arteriographic, histological signs as well as diagnostic criteria are restated. The pathogenesis is still unknown; a genetic predisposition and an auto-immune process could be involved. The treatment, presently symptomatic, cannot arrest the evolution which is often mutilating, when the patient continues to smoke. An early diagnosis is possible in the presence of superficial phlebitis or Raynaud's phenomenon. PMID- 3043482 TI - Effect of aldosterone on 86Rb fluxes in cultured kidney cells (A6). AB - This study was designed to evaluate the relative contributions of hormone induced changes in active and passive K+ transport in an epithelial cell line in continuous culture derived from toad kidney (A6) using 86Rb as a tracer for measuring unidirectional K+ fluxes. The effects of 24 h exposure to aldosterone (A) and aldosterone plus insulin (A+I) on unidirectional K+ fluxes were evaluated under short-circuited conditions and under open circuit conditions. In epithelia exposed to A, a small but significant amount of active K+ secretion was found, although it was not significantly greater than in control epithelia. The bidirectional fluxes in both A and A+I treated epithelia, under short-circuited conditions, increased by a similar amount over control values indicating an increase in apparent permeability of passive transepithelial K+ transport. Under open circuit conditions, A stimulated net K+ transport by about 5-fold over controls. The increase in K+ secretion produced by A under open circuit conditions could be explained by the combined effects of an increase in transepithelial K+ permeability and an increase in the transepithelial electrical potential difference (PD). The presence of I produced no additional effects to that of A on K+ transport under the conditions used in this study. It is concluded that the substantial increase in K+ secretion induced in A6 cells by 24 h exposure to A is primarily passive in nature. It is possible that the changes in both PD and transepithelial K+ permeability, which can account for the observed increase in K+ secretion, are secondary to the stimulation of active Na+ transport. PMID- 3043483 TI - Sixty-first president of APS Aubrey E. Taylor. PMID- 3043484 TI - Effects of chronic infusion of lipopolysaccharide on food intake and body temperature of the rat. AB - Unrestrained male Sprague-Dawley rats were infused for seven days with a low (2.45 micrograms/hr) or high (9.81 micrograms/hr) concentration of E. coli lipopolysaccharide (LPS). Compared to control (saline-infused) rats, food intake in the LPS-infused rats remained depressed for the entire infusion period. Despite this long-term suppression of food intake, fever was observed only during the daytime hours for the first two days of infusion. No significant increase in nighttime body temperature was observed. These data indicate that although tolerance to LPS occurred in rats with regard to its fever-inducing effect, tolerance with respect to its anorexigenic action did not occur. PMID- 3043486 TI - Medicinal plant research: 1953-1987. PMID- 3043485 TI - The antimalarial activity of Spathodea campanulata stem bark extract on Plasmodium berghei berghei in mice. PMID- 3043487 TI - Topical analgesia for the cutting of split-skin grafts: a multicenter comparison of two doses of a lidocaine/prilocaine cream. AB - The topical analgesic effect of two doses of a local anesthetic cream (EMLA, Astra) in the harvesting of split-skin grafts was compared in a double-blind multicenter trial. A standardized area of 200 cm2 at the donor site of 78 patients was randomly treated with 30 or 60 gm of cream 2 to 5 hours before the operation. There was no difference in pain between the groups (p = 0.53). In each group, 92 percent of the patients rated the pain as either none or slight, and 8 percent rated it as either moderate or severe. In conclusion, with the application times of 2 to 5 hours used in this trial, a dose of 15 gm EMLA cream per 100 cm2 is sufficient to provide analgesia for the cutting of split-skin grafts. PMID- 3043488 TI - Prevention of the teardrop areola following the inferior pedicle technique of breast reduction. AB - Elevation of the inferior portion of the areola with placement of dermal traction sutures will reduce the risk of an inverted teardrop areola deformity following inferior pedicle reduction mammaplasty. The long-term results in creation of a circular areola have been uniformly successful. PMID- 3043489 TI - Mr. Job van Meekeren (1611-1666) and surgery of the hand. AB - Job van Meekeren of Amsterdam was a surgeon, respected by outstanding contemporary medical doctors for his knowledge of medical literature and his skills, who made a definite link between anatomy and surgery. He showed a great interest in hand surgery, and interesting is a demonstration of flexor tendon repairs on corpses by one of his pupils. It is still a great joy to read his book today, which also gives a good representation of the state of the art of surgery in the seventeenth century in Amsterdam (Fig. 11). Names and addresses of patients are fully mentioned, so even today we know exactly where they lived and where the events took place. On the other hand, we also know quite well what the surgeons and doctors looked like through the efforts of many excellent painters who depicted anatomy lessons. In Amsterdam, barber-surgeons' guilds were very eager to sit for group paintings, centered around the teaching medical doctor (Table I). The painter Aert Pietersz in 1603 painted Dr. Sebastiaan Egberts surrounded by 29 surgeons, and in 1619, Dr. Egberts was painted once more, this time with five learning surgeons, by Thomas de Keyzer. Nicolaes Eliasz, named Pickenoy, painted Dr. Johan Fonteyn in 1625, and Rembrandt is well known for the Anatomy Lesson of Dr. Tulp (1632) and Dr. Deyman (1656). It is peculiar that a portrait of van Meekeren could not be traced. PMID- 3043490 TI - Gastroscopy: a primary diagnostic procedure. AB - EGD, using 1986 models of either the fiberoptic gastroscope or the videoscope, is a safe and accurate procedure that can be performed by any physician trained in the technique of endoscope passage. It may be performed at large medical centers or small rural hospitals, outpatient clinics, or even private offices. Patients themselves have indicated preference for endoscopic evaluation over the double contrast barium meal after they have experienced both procedures. The short time of procedure, its accuracy, safety, and its relative lack of discomfort to the patient lend it readily to being an initial component in the primary evaluation of symptoms of abdominal distress, gastrointestinal bleeding, dysphagia, esophageal reflux, persistent vomiting, and odynophagia. It is essential in the evaluation of complications of esophageal reflux and the evaluation of abnormal radiological findings in the upper gastrointestinal tract. It should never be overlooked in evaluating the patient with iron deficiency anemia of unknown etiology. Economic pressures have already moved EGD from the surgery wards to endoscopy labs and to the outpatient setting. These same forces will project more physicians into the role of the diagnostic endoscopist and the patient will benefit by decreased medical costs, quicker diagnosis and treatment, and enhanced continuity of care. PMID- 3043491 TI - Diverticular disease of the colon. AB - Diverticular disease of the colon has been called a "deficiency disease of the Western Civilization" and has become increasingly more common in industrialized countries during this century. Diets low in fiber predispose the patient to the development of this condition, and adding fiber to the diet is effective in prevention and treatment. Most patients with the condition are asymptomatic, but many patients experience pain and bowel disturbance along with other symptoms. Complications include infection, bowel obstruction and bleeding, and they may be life threatening. Medical and/or surgical management are indicated. PMID- 3043492 TI - Bacterial diseases of the colon. AB - Acute diarrhea of bacterial origin is discussed for seven enteric pathogens, and specific antimicrobial therapy based on positive identification is stressed. Selection of patients with self-limited disease not requiring antimicrobial therapy is emphasized in order to avoid costly laboratory tests. The role of daycare facilities in the spread of enteric pathogens in this country is discussed. This article includes a review of newer methods for treating infants and children with oral rehydration and rapid refeeding. PMID- 3043493 TI - Reflux esophagitis. Diagnosis, pathophysiology, and management. AB - Reflux esophagitis is a common disorder in which esophageal inflammation is caused by the reflux of gastric contents. The diagnostic approach includes documentation that reflux is present, that the patient's symptoms are caused by the reflux, and that esophageal mucosal damage has occurred. Therapy is guided by the current multifactorial pathophysiology model, which includes efficacy of the antireflux mechanism, volume of gastric fluid, potency of refluxed material, esophageal clearance, and tissue resistance factors. Although recurrences are common, treatment with life style changes supplemented with combinations of liquid antacids, H2 blockers, sucralfate, bethanechol, and metoclopramide is usually effective. PMID- 3043494 TI - Cholecystitis and cholelithiasis. AB - Cholelithiasis and cholecystitis, with their complications, remain major health problems in the United States. At this time, cholecystectomy is the treatment of choice for all patients with symptomatic gallstones and those with acute cholecystitis, except those who are too ill to undergo surgery. Present therapeutic options may be summarized as follows: Asymptomatic patients and those with flatulence and dyspepsia who have gallstones should be observed. Those who have symptoms of biliary pain, gallstone-induced pancreatitis, or common duct stones should have corrective surgery. Those who refuse surgery or who aren't surgical candidates might be treated with dissolution therapy. Dissolution of gallstones with chemical agents and extracorporeal shock-wave lithotripsy show some promise. We need a better understanding of the etiology and formation of gallstones to address the disease from a preventive standpoint and reduce the incidence of cholelithiasis and cholecystitis, and their complications. PMID- 3043495 TI - Viral hepatitis. AB - Knowledge of the etiologies and pathogenesis of viral hepatitis has exploded in the last two decades. Accurate serologic methods for identification of the causative agents have lead to an understanding of the importance of viral hepatitis in the pathogenesis of acute and chronic hepatitis, cirrhosis, and hepatocellular carcinoma. Although treatment of infection remains a challenge, accurate methods of diagnosis and prevention of infection are now at the physician's disposal. Successful control of these debilitating infections will depend upon the proper utilization of existing methods of prophylaxis. PMID- 3043496 TI - Gastroesophageal varices. Management of hemorrhage in the cirrhotic patient. AB - The appropriate management of the cirrhotic patient with bleeding gastroesophageal varices presents a continuing challenge for the primary care physician. These patients demand rapid assessment and skillful resuscitation. Confirmation of the suspected diagnosis should be sought by endoscopy. Endoscopic sclerotherapy currently is the treatment of choice for acute variceal hemorrhage. Unfortunately, despite prevention of recurrent hemorrhage, neither sclerotherapy nor portosystemic shunting has effected any significant alteration in the poor long-term survival of these patients. Newer approaches such as the use of propranolol offer some hope but doubtless will be hampered by the failure to alter the underlying liver disease. PMID- 3043497 TI - Acute pancreatitis. AB - Acute pancreatitis is most often secondary to prolonged excessive alcohol intake or biliary tract disease. The diagnosis is based on a combination of physical, laboratory, and radiologic findings. Differentiation from intra-abdominal processes that require surgical intervention is important. Treatment involves restoration of intravascular volume, correction of hypoxemia and metabolic derangements, and resting the gastrointestinal tract. Prognostic indicators are useful in identifying severe cases that may benefit from more aggressive monitoring, peritoneal lavage, antibiotic therapy, and surgical intervention. The recovery period may be complicated by sequellae of pancreatic necrosis and by sepsis. PMID- 3043498 TI - Upper gastrointestinal bleeding. AB - Bleeding in the upper gastrointestinal tract is a problem that confronts all physicians involved in primary patient care. It continues as one of the most common acute medical management problems, leads to substantial morbidity and mortality, and presents both diagnostic and therapeutic challenges to the physician. The purpose of this paper is to provide an overview of the common causes of upper gastrointestinal bleeding and to discuss initial assessment, diagnosis, management, and, where possible, prevention. PMID- 3043499 TI - Viral gastroenteritis. AB - As our ability to control many of the common infectious diseases has increased, attention has turned toward the less common or less severe infections. It is clear that worldwide, significant numbers of the cases of gastroenteritis in both adults and children are caused by viruses. Many of these viruses now are quite well understood and their control appears to be on the horizon. Many other etiologic agents are just being identified and will present a challenge to researchers and practitioners alike. PMID- 3043500 TI - Screening for colorectal cancer. Issues for primary care physicians. AB - Fecal occult blood testing, proctosigmoidoscopy, and digital rectal examination have been recommended as screening tests for colorectal cancer in asymptomatic people. This article evaluates the advisability of recommendations by applying accepted principles of screening to the case of colorectal cancer. PMID- 3043501 TI - Procedural skills in flexible sigmoidoscopy and colonoscopy for the family physician. AB - Procedural skills in flexible sigmoidoscopy and colonoscopy are widely accepted as diagnostic and therapeutic tools that assist physicians as they attempt to understand the biological and behavioral elements of undifferentiated gastrointestinal illness. Data support the ability of family physicians to perform and teach some of these endoscopic skills. Although one major contribution of these procedures is in the prevention of premature colorectal cancer mortality, other important patient care benefits are emerging for the office-based physician. Breadth of care enhances our most important therapeutic tool--the doctor/patient relationship. Unnecessary referral contributes to fragmentation and depersonalization of the health care system. Family physicians are urged to examine and acquire new procedural skills appropriate for their community. The process of privileges is discussed. PMID- 3043502 TI - Irritable bowel syndrome. Toward a biopsychosocial systems understanding. AB - The symptoms of irritable bowel syndrome (IBS) are usually a subset of a broader problem that meets DSM-III criteria for depression, anxiety disorder, somatization disorder, or adjustment disorder. A biopsychosocial perspective that addresses multigenerational family patterns of anxiety, depression, and somatization of stress suggests guidelines for understanding and treating patients with IBS symptoms. Effective treatment focuses primarily on helping patients cope with emotional disorders and psychosocial stressors, and secondarily on direct symptom relief. Psychotherapy is a valuable adjunct to medical treatment. The medications most likely to yield lasting benefits are the antidepressants. PMID- 3043503 TI - [Theodor Ziehen as psychiatrist and neurologist]. AB - With respect to other aspects of his extensive scientific work the essential features of the psychiatric and neurological contributions of Theodor Ziehen (1862-1950) are analyzed. PMID- 3043504 TI - [The practical value of the dexamethasone test in psychiatry]. PMID- 3043505 TI - [Various physiological parameters as predictive factors in the pharmacotherapy of endogenous depression]. PMID- 3043506 TI - [Clinical predictive factors in the pharmacotherapy of endogenous depression]. PMID- 3043508 TI - When does lucid dreaming become transpersonal experience? PMID- 3043507 TI - [The usefulness of projective methods in the diagnosis of schizophrenia]. PMID- 3043510 TI - Association for the Study of Dreams. International Dream Conference IV. June 1-7, 1987, Arlington, Virginia, U.S.A. PMID- 3043509 TI - Recent advances in child and adolescent psychiatry. PMID- 3043511 TI - Butterflies and bug hunters: reality and dreams, dreams and reality. PMID- 3043512 TI - From modernism to post-modernism: some implications for a depth psychology of dreams. PMID- 3043513 TI - The morning after: a pragmatist's approach to dreams. PMID- 3043514 TI - Dreams during language learning: when and how is the new language integrated. PMID- 3043515 TI - Metaphors of communication in the dreams of deaf people. PMID- 3043516 TI - Manifest content in dreams of Gussi and U.S. females: social and sexual interactions, achievement and fortune. PMID- 3043517 TI - Dream work with the mentally retarded. PMID- 3043519 TI - Olfactory stimuli and their effects on REM dreams. PMID- 3043518 TI - Characteristics of nightmare subjects and their nightmares. PMID- 3043520 TI - Group mythology and group development in dream sharing groups. PMID- 3043521 TI - [Social networks in schizophrenic disorders--a review]. AB - In the fifties the "social network"-concept was developed in order to improve the analysis of the structure of different social groups; later on, its emphasis has shifted to the study of "personal networks", that deal with the structure of the social relations of a "focal individual". In the latter sense the term "social network" will be used in this paper. Social networks are discussed in terms of morphological characteristics such as the size or the number of clusters as well as in terms of interactional characteristics such as the number of social contacts of the focal person. Studies concerning the social networks of schizophrenics that have been carried out so far showed the following results: In comparison with the social networks of the mentally healthy those of schizophrenic patients are markedly smaller in size and contain a smaller number of clusters. The proportion of family members in the social networks of schizophrenics is higher than in those of the mentally healthy. Social relations of the latter generally show a more complex structure than those of the schizophrenics. The social networks of schizophrenics patients with multiple admissions are generally smaller than those of first admission schizophrenic patients who also have more extrafamilial contacts. -The last section of this paper discusses methodical problems of the existing studies that deal with the social networks of schizophrenic patients; some suggestions are made concerning the application of the network-concept to psychiatric therapy. PMID- 3043522 TI - [The value of the social service unit in ambulatory gerontopsychiatric management]. AB - Care of aged mentally diseased patients is one of the subjects whose reform had been underlined as particularly urgent in the 1975 report on the state of psychiatry in the Federal Republic of Germany. Whereas in psychiatric hospitals there is now evidence of a growing trend to reduced hospital stay and to readmission after a period of release, no change seems to be in sight in respect of caring for the aged on an outpatient basis. The financial losses suffered by public health care organisations for aged mental outpatients have resulted in housing an increasing number of mentally ill persons of old age in homes for the aged, instead of caring for them as psychiatric outpatients. Since more and more aged persons will be represented among the population in future, it is more than ever before imperative to arrive at suggestions for a feasible solution. The article reviews the present situations (including acceptance of existing institutions by the aged) and explains the importance of public social care units in outpatient care of elderly people. The assessment was made by means of an official questionnaire operation in the Land of Baden-Wurttemberg. Efforts are suggested in respect of revised basis rules of organisation and staff to improve public outpatient gerontopsychiatric care. PMID- 3043524 TI - The child victim as witness in sexual abuse proceedings. AB - The psychiatrist working with a child witness in a sexual abuse case is at an intrinsic disadvantage. Most of the activities take place in a legal system where the psychiatrist may feel like a stranger in a strange land. Further, a major area of concern to the courts is cognitive development and memory, which unlike phenomenology, dynamics, and therapeutics, are somewhat remote from the psychiatrist's core expertise. It is not surprising, then, that the stress associated with the role of child advocate or expert witness often produces anxiety for the psychiatrist or may result in a frank avoidance of forensic tasks. This paper is intended to militate against such a reaction by familiarizing the psychiatrist with the salient issues of the sexually abused child as a witness. PMID- 3043523 TI - [The depression ward--a review of the current status]. AB - New therapeutic concepts are usually developed and realised step by step. Over the past years changes occurred mainly in two ways. There is a trend to reduce the size of the psychiatric hospital as such and/or to section off certain areas by introducing specialised units. From the experience with the so called "Affective Disorder Units" or "Mood Clinics" developed the concept of a Depression Unit, first introduced in Europe for research purposes at the Psychiatric University Hospital in Basle in 1968. In Germany the first Depression Unit was established at the PLK Weissenau in 1976. Similar units followed over the years at the PLK Reichenau, Weinsberg, the BKH Gunzburg, the Landesklinik Nordschwarzwald Hirsau/Calw and the Rheinische Landesklinik Bedburg-Hau, other hospitals are planning them. Existing units are described in terms of their structural organisation, staffing situation and the service they provide. The concept of the Depression Unit according to criteria laid down agreed upon in the literature is summed up, and emotional atmosphere, structural organisation, activation, and individual therapeutic measures, discussed. Advantages are mentioned such as better understanding of patients, allowing them to let go and be cared for, aimed regression, the effects of being taken along, improved communication, and a way of dealing with suicidal thoughts and depressive behavior that reduces anxiety. Disadvantages such as the danger of spoiling patients or inducing a sense of resignation both in patients and staff are discussed. PMID- 3043525 TI - Artifactual and genuine relationships of lateral difference scores to overall accuracy in studies of laterality. PMID- 3043526 TI - Excuses: their effective role in the negotiation of reality. PMID- 3043527 TI - Self-defeating behavior patterns among normal individuals: review and analysis of common self-destructive tendencies. PMID- 3043528 TI - Relapse after recovery from unipolar depression: a critical review. PMID- 3043529 TI - Psychosocial functioning and depression: distinguishing among antecedents, concomitants, and consequences. PMID- 3043530 TI - Is visual imagery really visual? Overlooked evidence from neuropsychology. PMID- 3043532 TI - Cross-validation of modal responses on the Children's Form of the Rosenzweig Picture-Frustration Study. PMID- 3043531 TI - Psychophysiological aspects of caffeine consumption. PMID- 3043534 TI - [The depth of depth psychology and the history of the individual]. PMID- 3043533 TI - Effects of methylphenidate on stimulus evaluation and response processes: evidence from performance and event-related potentials. PMID- 3043535 TI - [Dreams of children]. PMID- 3043537 TI - Phenomena at the advancing ice-liquid interface: solutes, particles and biological cells. AB - Ice formation in aqueous solutions and suspensions involves a number of significant changes and processes in the residual liquid. The resulting effects were described concerning the redistribution of dissolved salts, the behaviour of gaseous solutes and bubble formation, the rejection and entrapment of second phase particles. This set of conditions is also experienced by biological cells subjected to freezing. The influences of ice formation in that respect and their relevance for cryopreservation were considered as well. A model of transient heat conduction and solute diffusion with a planar ice front, propagating through a system of finite length was found to be in good agreement with measured salt concentration profiles. The spacing of the subsequently developing columnar solidification pattern was of the same order of magnitude as the pertubation wavelengths predicted from the stability criterion. Non-planar solidification of binary salt solutions was described by a pure heat transfer model under the assumption of local thermodynamic equilibrium. The rejection of gaseous solutes and the resulting gas concentration profile ahead of a planar ice front has been estimated by means of a test bubble method, yielding a distribution coefficient of 0.05 for oxygen. The nucleation of gas bubbles has been observed to occur at slightly less than 20-fold supersaturation. The subsequent radial growth of the bubbles obeys a square-root time dependence as expected from a diffusion controlled model until the still expanding bubbles become engulfed by the advancing ice-liquid interface. The maximum bubble radii decrease for increasing ice front velocities. The transition between repulsion and entrapment of spherical latex particles by an advancing planar ice-front has been characterized by a critical value of the velocity of the solidification interface. The critical velocity is inversely proportional to the particle radius as suggested by models assuming an undisturbed ice front. The increase of the critical velocity for increasing thermal gradients shows good agreement with a theoretically predicted square-root type of dependence. Critical velocities have also been measured for yeast and red blood cells. The effect of freezing on biological cells has been analyzed for human lymphocytes and erythrocytes. The reduction of cell volume observed during non-planar freezing agrees reasonably well with shrinkage curves calculated from a water transport model. The probability of intracellular ice formation has been characterized by threshold cooling rates above which the amount of water remaining within the cell is sufficient for crystallization.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3043536 TI - Cryo-electron microscopy of vitrified specimens. PMID- 3043539 TI - A description of hydatidiform mole with ultrasound case studies. PMID- 3043538 TI - A cytogenetic comparison of the responses of mouse and human peripheral blood lymphocytes to 60Co gamma radiation. AB - Experiments were conducted to compare the chromosome damaging effects of 60Co gamma radiation on mouse and human peripheral blood lymphocytes (PBLs). Either whole blood or isolated and pelleted mononuclear leucocytes (MNLs) were irradiated with a 60Co unit to yield exposures of 1, 2, 3, or 4 Gy. In addition, mice were whole-body irradiated in vivo with the same doses so that an in vitro in vivo comparison could be made. The results indicate that mouse PBLs irradiated in whole blood, whether in vivo or in vitro, respond similarly to 60Co gamma rays as measured by dicentric chromosome formation. In addition, mouse and human PBLs showed a similar radiosensitivity, but because the mouse PBL data were best fitted to an exponential function and the human PBL data to a quadratic function, direct comparisons were difficult to make. Pelleted MNLs from mice were much less sensitive to the clastogenic effects of gamma radiation than whole blood. This is believed to be due to hypoxic conditions that developed during irradiation and transport. Human PBLs did not show a marked difference whether irradiated in whole blood or as pelleted MNLs in tissue culture medium. PMID- 3043540 TI - Ephemeralization. PMID- 3043541 TI - Double-contrast upper gastrointestinal examination: technique and interpretation. PMID- 3043542 TI - Percutaneous aspiration and drainage of hydatid cysts in the liver. AB - Percutaneous aspiration of hydatid cysts of the liver was performed in 13 patients, and subsequent percutaneous drainage was performed in three of the 13. Aspiration was performed with ultrasound or computed tomographic guidance with 22 gauge to 19-gauge needles. Analysis of the aspirated specimen established the diagnosis of hydatid cysts in nine of the 13 patients. Fragments of the laminated membrane were seen in seven cases, scolices in two cases, and hooklets in two cases. In the four cases with negative results on aspiration, the diagnosis was established with surgical findings in one case and unequivocal immunologic results in three cases. In two patients, a mild allergic reaction with temporary pruritus was observed. In three patients, percutaneous drainage was performed with a 5-F to 8.3-F catheter, and sterilization of the cyst was achieved by injection of a scolicidal agent. No complications occurred at the time of drainage, and no recurrences developed during 6 months to 1 year after drainage. PMID- 3043543 TI - Transrectal US of the seminal vesicles and ejaculatory ducts: clinical correlation. AB - Transrectal ultrasonography (US) provides excellent anatomic detail of pathologic changes in the seminal vesicles and ejaculatory ducts. Fifty-two patients with US findings of seminal vesicle dilatation or cysts, ejaculatory duct cysts, or seminal vesicle or ejaculatory duct calculi were given questionnaires concerning a broad spectrum of genito-urinary symptoms. Compared with age-matched controls with normal US findings, patients with calculi in the seminal vesicles or ejaculatory ducts had a significantly increased prevalence of hematospermia and ejaculatory pain (P less than .01), and patients with cystic dilatation of the seminal vesicles were more likely to have perineal pain. Large midline cysts containing calculi or debris were symptomatic and probably represent mullerian duct remnants. Small cysts of the ejaculatory ducts were asymptomatic. Transrectal US may provide clinical insight into the causes of significant genitourinary symptoms that may previously have been ascribed to chronic nonbacterial prostatitis or have been considered to be idiopathic. PMID- 3043544 TI - US characteristics of frozen prostate. AB - Cryosurgery has previously been used successfully to treat prostatic carcinoma. Inability to monitor the freezing led to local complications that limited the use of this modality. In this animal study, monitoring of the freezing process was accomplished with real-time ultrasound. The margin of the frozen tissue appeared as a hyperechoic rim with posterior acoustic shadowing. The thawed cryolesions showed markedly decreased echogenicity compared with normal unfrozen prostate. These characteristic changes should allow safer and more efficacious application of prostatic cryosurgery. PMID- 3043545 TI - Transvaginal and transabdominal sonography: prospective comparison. AB - Transvaginal (TV) and transabdominal (TA) sonography were compared in a prospective study. A total of 230 examinations (126 pelvic, 104 pregnancy) were performed on 215 patients, ranging in age from 14 to 80 years. The improved anatomic detail on TV scans yielded new information in 138 (60%) examinations and better visualization of pelvic structures in 51 (22%) examinations. There was no important difference in diagnostic information provided by the two imaging modalities in 36 (16%) cases, and TV images were worse in five (2%). The clinical diagnosis was altered on the basis of TV sonographic findings in 54 (24%) cases and confirmed with certainty in 166 (72%). Diagnostic problems posed by TA scanning were not resolved by TV scanning in ten (4%) instances. Statistical analysis indicated that TV scanning was significantly better than TA scanning in the visualization of gestational sac contents (P less than .005), detection of fetal heart motion (P less than .001), and evaluation of the endometrial canal in the retroverted or retroflexed uterus (P less than .001). TV scanning was significantly better than TA scanning in visualization of the ovaries in patients with uterine leiomyomas (P less than .005) but not significantly better in peri- and postmenopausal patients (P greater than .05). PMID- 3043546 TI - Bone marrow imaging. PMID- 3043547 TI - Popliteal venous aneurysm. AB - Two new cases of popliteal venous aneurysm, confirmed with findings from venography, are added to seven previously reported cases revealed in the authors' search of the English-language literature. This rare anomaly usually shows as recurrent pulmonary emboli in patients with no underlying predisposition to deep venous thrombosis. Physical examination is usually not helpful in the diagnosis. Results of combined real-time and Doppler ultrasound should indicate the diagnosis, but venography is necessary for confirmation and further anatomic detail. Surgical treatment has been fraught with complications. Eight patients, including these two new cases, have undergone surgery, and none have had a recurrence of pulmonary embolism following surgery. PMID- 3043548 TI - The bird's nest inferior vena cava filter: progress report. AB - The bird's nest inferior vena cava filter, in clinical trial since 1982, has been placed in 568 patients at risk for pulmonary embolism. Of the 481 patients in whom the filter had been in place for 6 months or more, 440 were followed up clinically. The prevalence of clinically suspected recurrent pulmonary thromboembolism was 2.7% (12 patients) and that of inferior vena cava filter occlusion was 2.9% (13 patients). With the initial filter design, filter migration occurred in five patients. No migrations have occurred in the 147 patients treated with the filter after its modification to improve the anchoring system for greater stability. The bird's nest filter has proved safe and effective in the prevention of pulmonary embolism. PMID- 3043549 TI - Primary thyroid lymphoma: evaluation with CT. AB - The appearance of primary thyroid lymphoma on computed tomographic (CT) scans and clinical data for 15 patients were analyzed. The CT appearances were classified into three types: 12 patients (80%) had a solitary nodule (type 1), two (13%) had multiple nodules (type 2), and one (7%) had a diffuse goiter (type 3). In 47% of cases, both lobes were involved. The tumors had a strong tendency to compress (80%) or infiltrate (53%) the surrounding structures. The frequency of calcification (7%) or necrosis (7%) was low. Most patients (87%) had a rapidly enlarging thyroid mass, and six (40%) complained of symptoms from obstruction. All patients had coexistent Hashimoto thyroiditis. In 13 of 15 patients (87%), a highly probable diagnosis of thyroid lymphomas was determined with CT and clinical findings. A staging workup with CT and clinical findings confirmed at biopsy will allow appropriate therapy and may lead to improved prognosis for patients with this condition. PMID- 3043550 TI - US-guided biopsy of neck masses in postoperative management of patients with thyroid cancer. AB - High-frequency (10-MHz) sonography demonstrated a cervical mass or lymphadenopathy, or both, during postoperative follow-up of 52 patients who had undergone surgery for thyroid cancer. Percutaneous biopsy with ultrasonographic (US) guidance was performed in all 52 masses, 44 of which were nonpalpable. Malignant cells were obtained in 29 biopsies, and the results of 20 biopsies were negative, yielding benign lymphocytes only. Results in three biopsies were nondiagnostic due to hypocellular specimens. Therefore, 94% of biopsy results (49) of 52) were confidently assigned as either positive (56%) or negative (38%) for malignancy. There were no complications. High-frequency sonography can demonstrate clinically occult thyroid bed tumor recurrence and lymph node metastases. US-guided biopsy is an accurate and safe technique to confirm or exclude malignancy in patients at high risk of recurrence of thyroid cancer. PMID- 3043551 TI - Neonatal ovarian cysts: sonographic-pathologic correlation. AB - The authors reviewed the prenatal (11 infants) and postnatal (17 infants) sonograms and the clinical, surgical, and pathologic findings in 17 infants with an ovarian cyst to determine the sonographic features and natural history of neonatal ovarian cysts. An uncomplicated cyst (nontwisted, nonhemorrhagic) was completely anechoic and the cyst wall was imperceptible with sonography (five cases). A twisted or hemorrhagic cyst was cystic with a fluid-debris level, cystic with a retracting clot, septated with or without internal echoes, or solid (12 cases). These complicated cysts contained liquid and/or organized hematoma. Eleven of the 12 complicated cysts had a thin, highly echogenic wall. Cyst torsion commonly occurred in utero and could be diagnosed on prenatal sonograms by a typical sonographic appearance (eight cases). All of these infants were asymptomatic after birth. Four infants with hemorrhagic or twisted cysts were symptomatic. All cysts except one that resolved spontaneously were treated surgically, including three twisted cysts that showed no change in size over a 1 8-month interval. All of the cysts were of follicular origin. PMID- 3043552 TI - Robert Maxwell. A giant of the 20th century. Commemoration of Robert Maxwell, his 65th birthday and the 40th anniversary of Pergamon Press. PMID- 3043553 TI - Epinephrine in mammalian brain. AB - 1. Epinephrine is widely distributed in brains of various species throughout phylogeny but maintains its localization to hypothalamus and brainstem/medulla in all species studied. 2. A general decrease in brain epinephrine content is observed phylogenetically beyond fishes with wide variation within species. 3. The cellular localization of epinephrine forming enzyme is dissociated from epinephrine stores in hypothalamus where epinephrine appears to be primarily a hormone. 4. Three proposed functional pools of epinephrine are described. Synthesis of a hormonal pool and a second, perhaps nonfunctional, pool co-stored in noradrenergic terminals in the forebrain occurs extraneuronally and is probably inhibited acutely in the presence of high corticosteroids due to inhibition of uptake 2. Synthesis of epinephrine in the neuronal pool found primarily in the medulla may be enhanced due to increased PNMT activity in the presence of elevated corticosteroids. 5. Phylogenetic and pharmacological data suggest that epinephrine may play an important role in tonic regulation of the level of arousal, reward and sensitivity to environmental stimuli in mammals. PMID- 3043554 TI - Some aspects of stress and depression. AB - 1. The role of stress in depressive illness is discussed together with utility of the "learned helplessness" model and some neuropharmacological correlates of uncontrollable shock. 2. Similarities and differences between chronic antidepressant treatment and chronic stress treatment regimes are reviewed. 3. Finally the role of adaptive process in stress on antidepressant treatments is discussed. PMID- 3043555 TI - The effect of ceruletide on tardive dyskinesia: a double-blind placebo-controlled study. AB - 1. In a double-blind placebo-controlled study of 37 patients with tardive dyskinesia, the therapeutic effect of ceruletide was evaluated. 2. The patients were assigned at random to two groups that received either intramuscular injections of 0.8 micrograms/kg of ceruletide or placebo once weekly for 4 weeks. Conventional neuroleptic medication was not changed 3 weeks prior to and throughout the study period. Tardive dyskinesia was assessed using the Abnormal Involuntary Movement Scale over an 8-week period. 3. Ceruletide had a more pronounced effect on TD than the placebo however, because of the limited number of subjects examined, the difference between the two groups was not significant. Ceruletide was more effective than placebo in patients under 60 years of age (p less than 0.05) and whose antipsychotic medication was mainly butyrophenones. 4. No serious side effect was noted. 5. The findings suggested that ceruletide therapeutically benefits patients with tardive dyskinesia. PMID- 3043556 TI - [Clonal proliferation of hematopoietic stem cells and its relationship with leukemia]. PMID- 3043557 TI - [Biochemistry of manic-depressive disorder (I)--Aminergic transmission]. PMID- 3043558 TI - [Folding process of proteins--unfolding and refolding of the immunoglobulin fragments]. PMID- 3043559 TI - [Raman spectroscopic study of age- and cataract-related structural changes in the lens proteins of intact lenses]. PMID- 3043560 TI - [Maternal mRNAs that code for natural inducers of mesodermal structures in amphibian embryos]. PMID- 3043561 TI - [Host response against tumor in self defense mechanisms]. PMID- 3043562 TI - [Transcriptional regulation of MHC multigene family]. PMID- 3043563 TI - [Evaluation of monoclonal antibody to tumor-associated carbohydrate antigen for diagnosis of human cancer]. PMID- 3043564 TI - [Surgical treatment of cancer of the eyelids]. PMID- 3043565 TI - [Effect of ultraviolet irradiation on calcium-phosphate metabolism and vitamin D and 25-hydroxycholecalciferol levels in patients with chronic diseases of the digestive system]. PMID- 3043566 TI - [Mental disorders and suicide]. PMID- 3043567 TI - [The neutrophil system in alcoholics]. PMID- 3043568 TI - [Non-surgical methods of treatment of diseases of the anus and rectum]. PMID- 3043569 TI - Dosimetric considerations on multiple arc stereotaxic radiotherapy. AB - The aim of this paper is to present the physical and dosimetrical features of the stereotaxic radiosurgical method already published by the authors. This method concentrates the dose into the stereotaxic target volume, placed at the isocenter of a 4 MV X-ray beam. Computer calculations showed that: 9-17 arcs provide almost spherical dose distributions; the optimal photon beam quality is about 4-6 MV. The angle between adjacent rotation planes is 20-40 degrees and the arcs are 100 160 degrees wide. In this way the dose to healthy tissue is minimized. The dose distribution was experimentally verified both by ionometric and photodensitometric methods. The procedure for dose calculation at isocenter of fields as small as required by the radiotherapist, has been investigated. PMID- 3043570 TI - Effects of relaxation with guided imagery on surgical stress and wound healing. AB - The purpose of this experiment was to determine the effects of an audiotape series employing Relaxation with Guided Imagery (RGI) on the psychophysiologic stress response and wound healing in surgical patients. Twenty-four patients undergoing cholecystectomy were randomly assigned to either RGI or control (quiet period) conditions and measured against three indexes of recovery: state anxiety, urinary cortisol levels, and wound inflammatory responses. An analysis of variance for repeated measures revealed that the RGI group demonstrated significantly less state anxiety, lower cortisol levels one day following surgery, and less surgical wound erythema than the control group. Thus, the RGI tapes demonstrated stress-relieving outcomes closely associated with healing. PMID- 3043572 TI - The assessment of mood and affect in developmentally disabled children and adolescents: the Emotional Disorders Rating Scale. AB - There is an unmet need for a reliable method of evaluating disorders of mood and affect in developmentally disabled children and adolescents. Such a measure is required for both accurate diagnosis and treatment monitoring in this population. An extensive review of existing assessment techniques confirms that: (a) current techniques for the evaluation of emotional disorders in cognitively normal individuals are inappropriate for most children with developmental disabilities; and (b) current instruments designed for the assessment of developmentally disabled children pay inadequate attention to affective symptoms. In this paper, the preliminary version of a new instrument, the "Emotional Disorders Rating Scale for Developmental Disabilities" (EDRS-DD), designed to evaluate mood and affect in children and adolescents with developmental disabilities, is presented. A pilot study indicates that interrater agreement is good. PMID- 3043571 TI - [Clinical effectiveness and systemic toxicity of various mixtures of prilocaine and bupivacaine in axillary plexus block]. AB - The presently existing local anesthetics (LA) do not guarantee a rapid onset and simultaneously a long duration of action. The combination of a medium-long acting LA with bupivacaine, a long-acting LA with slow onset, could be means to achieve these aims. Prilocaine was chosen as the medium-long acting LA because it has the lowest toxicity of this group and for pharmacological reasons. METHODS. In a prospective, controlled double-blind study 100 patients scheduled for axillary block for elective surgical procedures of the hand or wrist were randomly assigned to five groups. Twenty patients in each group received either 40 ml prilocaine 1.5%; 40 ml bupivacaine 0.375%; 20 ml prilocaine 1% + 20 ml bupivacaine 0.5%; 20 ml prilocaine 2% + 20 ml bupivacaine 0.5%; or 20 ml prilocaine 2% + 20 ml bupivacaine 0.375%. The LA mixtures were freshly mixed 15 min prior to the axillary block. The blocks were performed using an immobile, short-beveled needle by anesthesiologists who were familiar with this technique. Analgesia was classified using the pin-prick method with 0 = no analgesia, 1 = analgesia, 2 = anesthesia. Motor blockade was classified with 0 = no motor block, 1 = paresis, 2 = paralysis. The following nerves were analyzed: ulnar, radial, median, musculocutaneous, and medial antebrachial. In 6 patients of each group plasma levels of the LA were measured by gas chromatography and methemoglobinemia was determined. Statistical analysis of the data was performed using the Student t-test and chi-square test on a level of significance of P less than 0.05. Results. All surgical procedures could be performed as planned in regional anesthesia. Twenty minutes after injection of the LA only 15% of the blocks were sufficient in the bupivacaine group, while in the other four groups 40%-50% of the blocks were complete (P less than 0.05). The degree of analgesia was deeper in the groups with 2% prilocaine and prilocaine alone than in the group with 1% prilocaine. Forty minutes after injection there were no significant differences between the groups. Motor blockade after 20 min was significantly lower in the bupivacaine group than in the prilocaine group (P less than 0.05). After 4 h all three prilocaine-bupivacaine mixtures showed a significantly more pronounced analgesia of the median nerve than the prilocaine group (P less than 0.02 0.001).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3043573 TI - Reasons for job loss: a review of employment termination studies of mentally retarded workers. AB - The current emphasis on employment preparation for mentally retarded people requires that more attention be focused on job retention. Knowledge of the reasons why mentally retarded workers were fired from their jobs could contribute to that effort. This article reviews studies over a 35 year period that have examined job loss in mentally retarded workers. First, reasons for job loss that relate to workers' vocational competence were separated from other reasons for job loss, such as economic and family factors. Then, reasons relating to vocational competence were classified into four categories: job responsibility, task-production skills, task-related social skills, and personal-social skills. Reasons for job loss among mentally retarded workers are broadly distributed across both social and nonsocial domains, including job responsibility factors such as attendance and punctuality, task-production factors such as inadequate quality and rate of work, task-related social skill deficits such as not following instructions and responding inappropriately to criticism, and personal social inadequacies such as inappropriate dress, bizarre and aggressive behavior, and inadequate or offensive verbal repertoires. The results of the review are discussed in relation to training issues for job retention of mentally retarded workers. PMID- 3043574 TI - [Value of 2-dimensional echocardiography in the evaluation of cerebral ischemia pathology in young adults]. PMID- 3043575 TI - [Renal function in multiple myeloma. Critical analysis of its prognostic value]. PMID- 3043577 TI - [Critical evaluation of the means of diagnosing cancer of the lung]. PMID- 3043578 TI - [Algorithms in spontaneous pneumothorax]. PMID- 3043576 TI - [A young woman with antecedents of nephropathy and changes in hepatic biochemistry]. PMID- 3043579 TI - [Beta 2 microglobulin and systemic lupus erythematosus]. PMID- 3043580 TI - [Eikenella corrodens empyema]. PMID- 3043581 TI - [Serum and urinary beta-2 microglobulin in cirrhotic patients]. PMID- 3043582 TI - The value of preoperative radiotherapy in esophageal cancer: results of a study by the EORTC. PMID- 3043583 TI - Interim analysis of a double-blind phase-III clinical trial of adjuvant levamisole versus control in resectable Dukes-C colon cancer: a study of the EORTC Gastrointestinal Tract Cancer Cooperative Group. PMID- 3043584 TI - The Northwest of England Rectal Cancer Trial. PMID- 3043585 TI - Five-year results of a prospective and randomized study: experience with combined radiotherapy and surgery of primary rectal carcinoma. PMID- 3043586 TI - The true role of external-beam irradiation in the initial treatment of cancer of the rectum. PMID- 3043587 TI - Combined-treatment approaches in the management of rectal cancer. PMID- 3043589 TI - Current therapeutic concepts in management of carcinoma of the anal canal. PMID- 3043590 TI - Approach of the treatment of colorectal liver metastases. PMID- 3043588 TI - Preoperative radiotherapy and radical surgery as combined treatment in rectal cancer. PMID- 3043591 TI - Controlled clinical trial for the treatment of patients with inoperable esophageal carcinoma: a study of the EORTC Gastrointestinal Tract Cancer Cooperative Group. PMID- 3043592 TI - 5-Methyltetrahydrofolic acid (MFH4): an effective folate for the treatment of advanced colorectal cancer with 5-FU. PMID- 3043593 TI - Intraoperative radiotherapy in carcinoma of the stomach and pancreas. PMID- 3043594 TI - Intraperitoneal chemotherapy and immunotherapy. AB - 1. The predictive model is validated empirically; a kinetic advantage for i.p. administration of certain antineoplastic drugs exist. 2. The concentrations achieved in the peritoneum are orders of magnitude greater than those found in the plasma, so rate-limiting systemic toxicities may be avoided. 3. The aspirin principle--if one is good, two are better--may not be applicable unless we can utilize this advantage to improve survival. PMID- 3043596 TI - The effect of adjuvant chemotherapy in gastric carcinoma is dependent on tumor histology: 5-year results of a prospective randomized trial. PMID- 3043595 TI - Phase-III clinical trial of adjuvant FAM2 (5-FU, adriamycin and mitomycin C) vs control in resectable gastric cancer: a study of the EORTC Gastrointestinal Tract Cancer Cooperative Group. PMID- 3043597 TI - Combined 5-fluorouracil and radiation therapy following resection of locally advanced gastric carcinoma: a 5-years follow-up. PMID- 3043598 TI - The value of a multidisciplinary approach in the management of gastric cancer. PMID- 3043599 TI - Biliary duct cancer: therapeutic nihilism or prospect. PMID- 3043600 TI - Treatment of pancreatic carcinoma: therapeutic nihilism? PMID- 3043601 TI - Adjuvant portal infusion chemotherapy in colorectal cancer. PMID- 3043603 TI - Effect of pH changes on the killing of Staphylococcus aureus and other mastitis pathogens by bovine neutrophil granule extracts. AB - A partly purified extract of granules from bovine neutrophils was used to investigate killing of mastitis pathogens by the non-oxidative killing system of the neutrophil. Eight strains of Staphylococcus aureus, two of Escherichia coli and two of Streptococcus uberis were used. Different strains of bacteria had different sensitivities to killing by the extract, three strains of staphylococci being particularly resistant. Whereas E coli were killed most effectively at pH 6.0, little or no killing of S aureus took place below pH 6.5 and, although S uberis were also killed better above neutral pH, significant killing also took place at acidic pH levels. Resistance to killing by the extract was not significantly correlated with that by intact neutrophils. PMID- 3043602 TI - [Need for social psychiatric services--a management model in scientific evaluation]. AB - Social-psychiatric services have for a long time been a focus in the controversial discussion about the further development of outpatient mental health care. Presented is the Baden-Wurttemberg Social Psychiatric Services model, which is oriented towards outpatient social welfare provision of persons with chronic mental illness, complementing the medical-psychiatric treatment provided by office-practice psychiatrists. Evaluation of the Social Psychiatric Service for its first year of full operation, 1984, has shown that this type of service delivery organization is capable, above all, of reaching persons with "recently" chronic mental illness, who are serviced in the framework of community mental health care systems mainly on an outpatient basis. Provision of care, i.e. of social welfare and counselling measures, may in this group constitute an efficient contribution towards their social rehabilitation. Effective functioning of this organizational model presupposes sufficiently well-established cooperation with other facilities and services involved in service provision, which, as the Baden-Wurttemberg trial has shown, can in actual fact be ensured. PMID- 3043604 TI - Applications of a cannulated extrusion needle during vitreoretinal microsurgery. AB - A specially designed cannulated extrusion needle facilitates the internal drainage of subretinal fluid during vitreoretinal microsurgery. Case histories demonstrate the use of this instrument in the management of complex retinal detachments including eyes with preretinal and subretinal hemorrhage, proliferative vitreoretinopathy with open peripheral retinal breaks, retinal detachment with giant retinal tear, and combined traction-rhegmatogenous detachments due to diabetic retinopathy. This method of internal drainage appears to have specific advantages over standard techniques by allowing the removal of subretinal hemorrhage, the utilization of pre-existing peripheral breaks for internal drainage, and the complete reattachment of shallow posterior pole retinal detachments to allow laser endophotocoagulation. PMID- 3043606 TI - [Glaxo Laboratories: 20 years of antibiotic therapy. The antibiotic, the germ and the patient]. PMID- 3043607 TI - Economic issues in the care of the elderly cancer patient. PMID- 3043605 TI - Lung clearance of 99mTc-DTPA aerosol in conscious sheep. AB - This study was initiated to determine the rate and characteristics of 99mTc-DTPA clearance from the whole lung in a group of 9 sheep. Submicronic aerosol droplets were delivered to unsedated sheep held in a sling-like frame. Best fits for clearance curves to single- and biexponentials were calculated. The monoexponential T50 for the aerosol clearance was 293 min +/- 74 SD. Background correction was found to have a minimal effect (approx. 10%). Biexponential fitting only marginally improved correlations in the 8 healthy sheep, but in one additional animal with clinical evidence of pneumonia, clearance was faster and biexponential fitting was substantially better. These clearance values in conscious sheep are longer than previous findings with 99mTc-DTPA in anesthetized sheep. There appears to be a wide variation of radioaerosol 99mTc-DTPA lung clearances among different species, sheep exhibiting a comparatively prolonged clearance profile. PMID- 3043608 TI - Cerebrospinal fluid analysis in the dog: methodology and interpretation. PMID- 3043609 TI - The cytologic examination of synovial fluid. PMID- 3043610 TI - The cytologic examination of body cavity fluids. PMID- 3043611 TI - The cytologic diagnosis of mesenchymal tumors. PMID- 3043612 TI - Advances in the cytologic diagnosis of canine lymphoma. PMID- 3043613 TI - [Bullosis in diabetics. Apropos of a new case]. AB - Idiopathic bullae occurring in diabetics constitute an uncommon condition which may reveal the diabetes, announce its decompensation or represent a mere epiphenomenon. The authors report the case of a woman who experienced two episodes of idiopathic bullae: the first one disclosed the diabetes, the second one heralded its decompensation. Both episodes were preceded by painful paraesthesia. The bullae contained a clear and serous fluid and the separation was intradermal. Direct immunofluorescence with anti-fibrin serum only showed fluorescence in the lumen of the bulla and in subjacent dermal areas. Electron microscopy demonstrated that the tissue separation was at the base of the stratum corneum. Other clinical cases and pathogenetic theories found in the literature are reported. In contrast with other bullous diseases occurring in diabetics and often of poor prognosis, idiopathic bullae is a benign condition requiring no more than a symptomatic treatment. PMID- 3043615 TI - [Involvement of the digestive tract in diabetes: manifestations, treatment and pathogenic hypotheses]. PMID- 3043614 TI - [Diaphragmatic paralysis disclosed by acute respiratory failure. Apropos of a case of amyotrophic lateral sclerosis and review of the literature]. AB - A case of acute respiratory failure revealing amyotrophic lateral sclerosis is reported. The other neurological diseases with diaphragmatic paralysis are recalled. PMID- 3043616 TI - [Clinical approach to eating behavior disorders in adults]. PMID- 3043617 TI - [Tuberculosis in the context of acquired immunodeficiency syndrome]. PMID- 3043618 TI - [Necrotizing form of Horton's disease. A case with quadruple mucous localizations]. PMID- 3043619 TI - [Pharmacological and molecular aspects of the regulation of eating behavior. With special reference to the role of catecholamines and effects of amphetamine]. AB - Among the numerous endogenous substances involved in the regulation of feeding behaviours, the catecholamines are in the front rank. The numerous studies devoted to this aspect of catecholamines emphasize the importance and complexity of their intervention. Depending on the cerebral structures on which they act and on whether noradrenaline or dopamine are concerned, orexigenic or anorexigenic effects have been described. Alpha-2 and beta adrenergic receptors as well as D1 and D2 dopaminergic receptors participate in these effects. Amphetamine, which is an indirect catecholaminergic agonist, mobilizes neuronal catecholamines and fosters their various effects. Moreover, it exercises direct effects by its association with sites borne by glycaemia-sensitive neurons. This target seems to be common to a wide variety of anorectic agents. They are thought to reproduce on this hypothalamic "glucostat" the effect of a high blood glucose level, thus triggering off signals of satiety. In this unifying hypothesis, the diverse pharmacological profiles these agents are known to possess would result from associated properties. PMID- 3043620 TI - [Health assessment by means of physical activity and nutrition]. AB - Physical activity and nutrition are the most important factors in preserving and/or improving health conditions. The relationship between exercise and nutrition is focused on two different directions: one is the direction toward health promoting, the other is for performance of a long heavy muscular work. For preserving and/or promoting health conditions, energy balance between intake and expenditure must be taken into account. The amount of glycogen stored in muscles is the essential factor to promote a long distance performance of working muscles. Topics that influence the energy balance and glycogen loading used by endurance athletes to improve performance are reviewed. PMID- 3043622 TI - Instrumental variables in the evaluation of diagnostic test procedures when the true disease state is unknown. AB - We explore the estimation of sensitivity and specificity of diagnostic tests when the true disease state is unknown. Instrumental variables which subdivide the patient population are used. A logistic model, relating these instrumental variables to the (unknown) true disease state is proposed. It is shown that this procedure allows the goodness-of-fit to the resulting model to be tested. PMID- 3043621 TI - The hidden effect of time. AB - It is customary to regard datasets as homogeneous with respect to the order of collection of the measurements. Examples are given in which this assumption is breached. Hidden time trends have implications for the design of studies, their analysis and interpretation. It is suggested that, if the order of observations is known, a plot by time should be performed, perhaps using a cusum. PMID- 3043623 TI - Variance estimation for epidemiologic effect estimates under misclassification. AB - This paper presents basic methods for construction of variance estimators for epidemiologic effect estimates when adjusting for misclassification. Methods are described for differential and non-differential misclassification, and for external and internal estimates of classification rates. The methods take account of sampling variability in both the observed data and the estimated classification rates. The methods are illustrated in a case-control analysis of the association of antibiotic use during pregnancy with sudden infant death syndrome. PMID- 3043624 TI - [Precursors of megakaryopoiesis in man. Current findings]. PMID- 3043625 TI - [Renal grafts at the Free University of Brussels after 25 years]. PMID- 3043626 TI - [Surgical treatment of pleural mesothelioma]. AB - Mesothelioma which is the primary tumor of the pleura has recently been subject to many researches because of its poor prognosis and the difficulties met in radical therapy. Between 1974-1986 total 46 patients with pleural mesothelioma were treated surgically at our center. Finally it was concluded that only the cases in stage I and with epithelial malignant pleural mesothelioma should be subjected to radical surgery. The remaining patients should be treated with conservative methods. PMID- 3043627 TI - [Sarcoidosis and autoimmunity. Apropos of 2 cases]. AB - Several immunologic abnormalities exist in patients with active sarcoidosis. Impairment of in vivo and in vitro cellular immunity is well known. Humoral immunity disorders are characterized by increase in polyclonal serum immunoglobulin, existence of circulating immune complexes and auto-antibodies. In the serum, these abnormalities are nearly the same than disorders observed during auto-immune diseases. We report 2 cases of patients where active sarcoidosis was associated with auto-immune disease. Then we discuss the relationship which could exist between the 2 types of diseases. The first patient was a woman (aged 66 years). She presented an active sarcoidosis and clinical symptoms of sclerodermia. The second patient, a man (aged 24 years), presented active sarcoidosis diagnosed 4 years ago. He had never received any treatment. Thrombopenic purpura occurred suddenly and needed steroid therapy. Antibodies to platelets were found in the serum. These associations raise some questions: 1) are they only fortuitous? such cases are rare but their frequency is perhaps underestimated; 2) could any common abnormalities of the immune response explain the emergence of the 2 diseases? 3) could one of the diseases favour emergence of the other? This hypothesis seems possible only if sarcoidosis is the initial disease. Eventually these associations also raise difficult therapeutic problems and must incite to explore precisely the immunity of patients with active sarcoidosis. PMID- 3043629 TI - [Thoracic metastases of ependymoma. Apropos of a case and review of the literature]. AB - A 27-year old woman with occipital ependymoma treated by surgery and radiotherapy for multiple recurrences developed a right lymphocytic pleural effusion. After 3 inconclusive needle biopsies of the pleura, pleurectomy was performed. Pathological examination with a specific immunohistochemical marker provided a diagnosis of pleuro-pulmonary metastases from the ependymoma. This rare case of extraneuraxial metastasis from a cerebral ependymoma is discussed in the light of published data. Its originality lies in that a positive diagnosis of pleural metastases was obtained with an immunohistochemical marker, the gliofibrillary antiprotein serum (anti-GFAP) specific of glial tissue. PMID- 3043628 TI - [Occupational bronchopulmonary pathology caused by woodworking: diagnostic approach]. AB - Woodworkers have a varied breathing pathology, with involvement of the upper airways mainly by ethmoidal cancers and rhinitis, tracheo-bronchitis with asthma, hypersensitivity pneumonitis due to wood moulds. The physiopathological mechanisms are numerous and often intricate: physical mechanism due to the irritant effect of wood dust, toxic mechanism caused by chemical substances which are present in wood dust or come from wood preservation products, immunological and allergic mechanism in which different allergens are concerned. Breathing pathology is the subject of 2 tables (No. 47 and 36) among those concerning occupational diseases. Lung function tests play an important role in the diagnosis of these diseases, with their 3 main objectives: search for airway obstruction, evaluation of bronchial hyperreactivity by means of non-specific challenge tests, and specific challenge tests performed in a laboratory or on the work site. PMID- 3043630 TI - [Physiological basis for the analysis of fetal heart rate]. PMID- 3043631 TI - [Neurologic manifestations in systemic lupus erythematosus]. PMID- 3043632 TI - [Indications and contraindications of peridural anesthesia in obstetrics. Review of the literature]. AB - With the help of an extensive review of the french and foreign literature, and personal statistics concerning 500 peridural anesthesias (PDA), the authors analyze the indications and contraindications of this type of anesthesia, in 1987. It is possible to differentiate well accepted indications: caesarean section, inducement, labor test, dynamic dystocia, delivery of fragile fetus, desire of the patient; definite contraindications: patient's refusal, coagulation disorders, emergency situations, some cardiopathies; debatable indications: breech delivery, scarred uterus, twin delivery, maternal medical problem, where each particular case must be evaluated. In his study, the authors demonstrate that indications tend to become broader and contraindications to become more infrequent. They stress a necessary co-operation between obstetrician and anesthetist to make this decision. PMID- 3043633 TI - [A new case of hydatidiform mole occurring on one of the ova of a twin pregnancy]. AB - The authors report here a new case of hydatid mole in one of the eggs of a mole pregnancy. 48 other cases have been reported in 13 publications. The birth of a viable, well-developed fetus is described in 13 instances (27%), fetal death in utero in 22 instances (46%), birth of a non-viable fetus 11 times (23%) and malignant degeneration twice (4%). Medical interruption of the pregnancy is indicated for the slightest sign of fetal anomaly or maternal complication. Prostaglandins may be used for that purpose. However, it is possible to let such a pregnancy follow its course. PMID- 3043634 TI - [Estrogen substitution using a transdermal system]. PMID- 3043635 TI - [Computerized tomography diagnosis of traumatic spinal changes: instability criteria, biomechanical fracture classification and possibilities for errors]. AB - This is a review of CT criteria in the evaluation of potential instability, giving a simple guideline to fracture classification according to biomechanics and a survey of imaging pitfalls, as well as a diagnostic regimen of radiological imaging for acute trauma and posttraumatic states. PMID- 3043636 TI - [Oncogenes and their role in human tumors]. PMID- 3043637 TI - [New biomodulator treatment, treatment with interferon alpha in hematology and treatment using interleukin 2]. PMID- 3043639 TI - Anorexia nervosa (AN). PMID- 3043638 TI - [Thrombolysis at home]. PMID- 3043640 TI - Kaposi's sarcoma. Then and now. Nodular lesion of palate as the only manifestation of the disease in a 70 year old heterosexual woman. AB - Kaposi's sarcoma is a transmissible disease, possibly associated with a virus, characterized by multiple pigmented lesions. Its epidemiology has dramatically changed in the last 4-5 years and has been the focus of considerable attention and controversy. PMID- 3043641 TI - [Maxillofacial manifestations of Cowden's disease. Apropos of 2 cases]. AB - Two new cases of Cowden's disease observed in two young sisters are reported. The diagnosis was based on the clinical observation of oral mucosa lesions (gingival hyperplasia, papillomatosis, scrotal tongue). In one case, visceral manifestations in childhood were observed. Cowden's disease is a familial affection (autosomal dominant trait) characterized by association of oral mucosa and skin lesions and of multiple hamartomas involving glandular tissues. Clinical and familial history investigations are justified by the high risk of malignant tumors (breast carcinoma). PMID- 3043642 TI - Verrucous carcinoma of the oral cavity. AB - The clinical course and morphologic (light and ultrastructural) characteristics of a case of verrucous carcinoma of the lower gingival mucosa are presented. Clinical course of the patient was unfavorable following surgery (supposedly incomplete) and radiotherapy (6,000 R) of the tumor. The therapeutic factors that may influence an unfavourable prognosis are commented upon, especially in regard to a slow-growing neoplasm with a generally good course. PMID- 3043643 TI - [Oral nevocellular nevus]. AB - This is a clinical report of a nevus cell nevus arising on the palatal mucosa. The incidence, the etiology, the differential diagnosis and the treatment of this mucosal pigmented lesion are reviewed. PMID- 3043645 TI - The effect of acoustic trauma on the tectorial membrane, stereocilia, and hearing sensitivity: possible mechanisms underlying damage, recovery, and protection. AB - The aim of the present investigation was to determine: 1) the relationship between changes in auditory sensitivity and alterations in stereocilia micromechanics and tectorial membrane morphology after acoustic overstimulation; 2) the rate of growth of a threshold shift in stereocilia following in vitro overstimulation; and 3) if the damaging effects of noise trauma can be reduced by pre-exposure to a low level acoustic stimulus. After exposure to a 1.0 kHz pure tone signal at 105 dB SPL for 72 hours, the threshold of the auditory brainstem response was broadly elevated by approximately 40-50 dB; the inner hair cell stereocilia became less stiff; and morphological alterations were observed in the middle zone of the tectorial membrane. The location of both the stereocilia and tectorial membrane alterations corresponded to the region of the cochlea demonstrating a threshold shift. Following a recovery period from overstimulation, the auditory brainstem response showed some improvement yet a 25 dB threshold shift remained. At this time, swelling of the afferent dendrites beneath the inner hair cells was observed together with scattered outer hair cell loss. Also, the inner hair cell stereocilia regained their normal stiffness characteristics. The in vitro experiments demonstrated that overstimulation reduced the stiffness of the inner and outer hair cell stereocilia bundles. A threshold shift increased systematically with exposure duration and intensity. After 6 minutes of overstimulation, the threshold shift exhibited a plateau whose magnitude was dependent upon the exposure intensity. Stereocilia micromechanics were shown to be dependent on the metabolism of the hair cell. The pre-treatment to a low level acoustic stimulus (81 dB SPL) prior to exposure to a stimulus known to yield a permanent threshold shift resulted in a 20 dB reduction in the threshold shift relative to the group not pre-exposed as well as complete recovery from the threshold shift after 2 months. PMID- 3043644 TI - [Sonography in diseases of the breast]. PMID- 3043646 TI - The OMGE acute abdominal pain survey. Progress report, 1986. AB - By 1986 the central analysis team of this on-going multinational survey had received a total of 10,682 cases for analysis and had accepted 10,320. In all, some 26 centres in 17 countries, involving over 200 doctors, had participated in this survey. A common protocol was used for data collection; around 98% of all possible data was recorded (using precirculated definitions) and analysed via a computer-aided system in Leeds, England. The construction and format of a series of computer-aided decision-support and teaching programs has been described in an earlier (1982) report. These programs are currently available/in use in 10 countries. The present report concentrates upon an update of current material collated for the survey, some demographic trends, and special subreports (as with IBD survey) concerning acute abdominal pain in children and elderly patients, together with some preliminary data on the value of leucocyte count in patients with suspected appendicitis. PMID- 3043648 TI - Do the stigmata of recent haemorrhage have additional prognostic value in patients with bleeding duodenal ulcer? AB - To assess the additional prognostic value of endoscopic stigmata of recent haemorrhage (SRH) in addition to clinical data, a pilot study was conducted on 207 duodenal ulcer patients from the OMGE survey. The incidence of SRH was compared in 145 patients who settled and 62 who re-bled. Only active bleeding emerged as significantly commoner in the latter group. Two computer-assisted predictions of further bleeding (before and after addition of the stigmata to a clinical database) were compared with the final outcome. Little improvement, in terms of prognosis for further bleeding, was obtained when SRH data were added. Similar findings were observed in a smaller series from Marburg, FRG. PMID- 3043647 TI - Acute abdominal pain in children. AB - This preliminary communication describes the initial results of a further special study investigating the disease spectrum and clinical presentation in a total of 1080 children admitted to hospital with acute abdominal pain (677 from the Children's Hospital, Sheffield, England, and the remaining 403 from hospitals in Paris, Oslo, Copenhagen, and Deventer). The disease spectrum in children differs radically from that in adults, well over 90% of cases being due to either acute appendicitis or non-specific abdominal pain (NSAP). The clinical presentation of both appendicitis and NSAP was found to differ from that in older age groups. These findings imply clearly that the use of the existing OMGE database for computer-aided diagnosis--using data drawn from cases of all ages--may not be optimal in children. A fresh database (using only children's data) was therefore created and tested. Its sensitivity for appendicitis was equivalent to that of inexperienced clinicians (79.6% versus 77.3%). The computer's specificity (over 70%) was higher than that of clinicians (52.7%). The findings also re-emphasise the value of disciplined data collection, and the implications for teaching are discussed. PMID- 3043649 TI - Use of a multinational survey to provide clinical guidelines for upper gastrointestinal bleeding in developing countries. AB - The OMGE multinational survey of patients with upper gastrointestinal bleeding demonstrated that it was possible to predict patient outcome, using a computer and a database of information from many centres. It remained to be seen whether this database could be used in specific remote areas of the world. To answer this question, two areas have been studied, Sikkim and China. In Sikkim, when the computer-aided prognostic system was used, 69% of patients put into a high-risk group for rebleeding actually did so; and 54% died. By contrast, only 2% of patients placed into a low-risk group for rebleeding did so. As there is little computer technology in Sikkim, a simplified scoring system was developed which gave the same predictive accuracy as the computer system. In China there was a slightly lower accuracy with both systems. Hence a new database and scoring system were created, using only information from Chinese patients. This database improved the results. From the studies it is suggested that these types of systems can be of value to patients in remote areas by targeting patients at high risk rebleeding or dying, so that the scarce resources available can be best used. PMID- 3043651 TI - Downhill esophageal varices caused by benign giant lymphoma. Case report and review of downhill varices cases in Japan. AB - Incidentally detected "downhill" varices of the entire esophagus in a 26-year-old man were found to be due to a benign giant lymphoma in the posterior mediastinum. The superior vena cava was not obstructed. Following resection of the tumor the varices disappeared. Substantial blood drainage to the esophageal veins was assumed to be the causal mechanism. PMID- 3043650 TI - Cryptococcus neoformans antibody levels in patients with AIDS. AB - Anti-Cryptococcus neoformans capsular polysaccharide (CPS) antibodies were measured by ELISA in patients with AIDS related complex or AIDS without a known history of cryptococcosis and in heterosexual healthy controls. Total and IgG anti-CPS antibody activity was rarely detected in patients, with mean levels lower than in controls, whereas IgM antibody activity was similar in the 3 groups. Since both humoral and cellular immunity appear to be of great importance during cryptococcosis, the inability of AIDS patients to synthetize specific IgG antibodies could impair an alternative host defence mechanism to cellular immunity. PMID- 3043652 TI - Current concepts in the assessment of effects of metals in chronic low-level exposures--considerations of experimental and epidemiological evidence. AB - Human exposure to metals and their compounds may give rise to a number of adverse effects. Of particular interest are such effects that can be elicitated by chronic low-level exposure, since such exposures may occur in human populations both in the general environment and occupationally. The adverse effect that occurs at the lowest exposure has been termed the "critical effect" since it is critical in relation to preventive action aiming at limiting exposures to a safe level. When reviewing the present evidence on the toxicology of metals it becomes evident that "critical effects" may be related to various organ systems, disease categories and biochemical mechanisms of damage. A category of effects, which is of particular interest in relation to critical effects, is hypersensitivity (e.g. with dermal or respiratory manifestations). However conceptual recognition of this category of effects is as yet incomplete. Other important considerations are whether effects display threshold- or non-threshold type dose-response relationships, the latter category obviously being more likely to display a low incidence response at low dose exposure. In this context mutagenic, carcinogenic, developmental and reproductive toxicity of metals are of particular interest. Information on these types of effects compiled during previous meetings of the scientific committee on the toxicilogy of metals and recently updated, indicates that some compounds of arsenic, beryllium, cadmium, chromium and nickel may be regarded as carcinogenic for humans. Available evidence in experimental systems as well as in humans concerning effects of metallic compounds on male and female reproductive systems, congenital teratogenicity and perinatal toxicity as well as effects on the developing central nervous system indicates that such effects should be considered further in relation to critical effects for arsenic, cadmium, lead, mercury and their compounds. PMID- 3043653 TI - Analytical procedures and clinical reference materials in monitoring human exposures to trace metals with special reference to Cr, Pb and T1. AB - This paper presents some special guidelines concerning sample collection and handling of clinical specimens for Cr, Pb and T1 analyses. The analytical methods used are briefly discussed. The available clinical reference materials (certified or not) for these three elements are reviewed and future developments are suggested. PMID- 3043654 TI - Environmental exposure to thallium. AB - This paper reviews recent data and findings concerning the sources of thallium in the environment and its current environmental levels and exposures, particularly with respect to human exposure. Special emphasis is given to some recent cases of environmental thallium pollution that were recognized around several cement factories in the Federal Republic of Germany. Contaminated food grown in these areas gave rise to a significantly increased thallium exposure of the population living there. Recent studies show that the thallium level in human urine normally is below 1 micrograms/g creatinine and that the normal concentration range of thallium in human air is approximately 5-10 ng/g. These data allow to recognize even minor undue exposure to thallium in the work and general environment. PMID- 3043655 TI - Review of occupational epidemiology of chromium chemicals and respiratory cancer. AB - Several epidemiologic studies have investigated the association between cancer risk and employment in chromium producing and using industries. Strong and consistent associations have been found between employment in the primary chemical producing industry and the risk for respiratory cancer. Workers employed in chromate pigment production and possibly spray painters of chromate pigment paints appear to be at excess risk of respiratory cancer. Chrome platers may also be at excess risk, although the evidence is limited. A few studies indicate that chromium alloy welding may also be an exposure source of concern. Some studies of ferrochromium alloy workers have shown an excess risk for respiratory cancer, although the risk may in part be due to concomitant exposures. The evidence indicates that the hexavalent form of chromium is the primary agent of chromium carcinogenesis. Solubility and other characteristics of chromium compounds may also play a role in determining risk. PMID- 3043656 TI - Chromium carcinogenicity; a review of experimental animal data. AB - Since 1932 exposure to various hexavalent chromium compounds has been known to constitute a cancer hazard to industrial workers. A number of animal studies have been performed and chromates have been administered by inhalation, intratracheally, by intravenous and subcutaneous injection, intraperitoneally and orally. The human epidemiological studies have provided convincing evidence that zinc chromate is a potent carcinogen and there is some evidence that calcium chromate and chromium trioxide also constitute a cancer hazard in humans. The human studies have not selectively confirmed carcinogenic potency for other chromium compounds. The animal studies confirm the carcinogenic potency of calcium chromate and zinc potassium chromate and present strong evidence that chromates of lead and strontium are carcinogenic in animals. The significance of the water solubility for the carcinogenic potency of the chromates has not been clarified by animal studies. PMID- 3043657 TI - Review of genetic effects and mechanisms of action of chromium compounds. AB - When similar experimental conditions are adopted to test Cr(VI) and Cr(III) genotoxicity, only the former gives positive results, and Cr(III) is scored as genetically inert. Since Cr(III) seems to be the main final genotoxic agent inside the cell, the observed difference in genetic activity of the two oxidation states can be referred to the easy transport of Cr(VI) across the plasma membrane, as opposed to the lack of membrane carriers for Cr(III). If the experimental design is adjusted to Cr(III) properties, Cr(III) appears (mildly) genotoxic. The accepted inactivity of Cr(III) on whole cells is thus questionable and the environmental impact of Cr(III) accumulation is worth reconsidering. PMID- 3043658 TI - Metabolic reduction of chromium as a threshold mechanism limiting its in vivo activity. AB - Various mechanisms tend to reduce hexavalent chromium (Cr(VI] in the organism, both outside and inside target cells. A reducing activity has been demonstrated in the blood (mainly in erythrocytes), in secretions of the alimentary tract (saliva, gastric juice) and in the lumen of terminal airways (epithelial-lining fluid and alveolar macrophages). Preparations of several types of cells from various animal species--including human liver, lung parenchyma and bronchial tree cells--are capable of metabolically reducing Cr(VI) to a variable extent. This process can be ascribed to different cytoplasmic components, e.g. to mitochondria, microsomal fractions and especially to cytosolic fractions, where reduction is partly due to electron donors (e.g. glutathione) and mainly to NADPH dependent enzyme activities, such as DT-diaphorase. Reduction is not only selectively stimulated by enzyme inducers but also, in rat lung cells, by the repeated intra-tracheal administration of Cr(VI) itself. All these reducing reactions are interpreted as detoxifying mechanisms, which constitute a threshold phenomenon in chromium carcinogenesis. PMID- 3043659 TI - Toxicokinetic aspects of thallium poisoning. Methods of treatment by toxin elimination. AB - The elimination techniques which are used in the treatment of thallium poisoning are evaluated in toxicokinetic perspective. Prussian Blue therapy and forced diuresis are effective in the treatment of acute as well as subacute thallium intoxications. Both elimination techniques can be applied without danger from neurological side effects. Generally, the combined use of these techniques is recommended. Since obstipation as well as renal impairment are common in thallium poisoning, extracorporal elimination techniques are of additional importance. In particular, charcoal haemoperfusion has proven to be successful in the elimination of thallium from the body, especially in the early phase of the intoxication. PMID- 3043660 TI - Haematological effects of lead. PMID- 3043661 TI - Isolation of dinitropyrene in emission gas from a municipal incinerator and its formation by a photochemical reaction. AB - In a previous paper, direct-acting mutagenicity was reported in emission gas from incomplete municipal incineration using Salmonella typhimurium TA-98 and TA-100. This paper reports the detection of dinitropyrene (DNP) as a direct-acting mutagen using nitrogen selective gas chromatography. The gas-phase photochemical reaction of pyrene with nitrogen dioxide was examined in a quartz vessel with various reaction times and temperatures. 1-Nitropyrene (1-NP) was readily formed from pyrene in the absence of light irradiation and at low temperature, but DNP was not formed under similar conditions. DNP formation was catalyzed by nitric acid. The formation of DNP is dependent on light irradiation, temperature and HNO3 as catalyst. In a combustion source these factors affect the formation of DNP. PMID- 3043663 TI - Marijuana test: no ibuprofen interference. PMID- 3043662 TI - Signal transduction and transcriptional regulation by glucocorticoid receptor LexA fusion proteins. AB - The glucocorticoid receptor regulates transcriptional initiation upon binding to its cognate hormone. A series of fusion genes was constructed to examine the mechanism of hormone-regulated transcriptional enhancement. The DNA binding domain of the bacterial LexA repressor was fused to receptor derivatives lacking the region that is necessary and sufficient for specific DNA binding and transcriptional enhancement at glucocorticoid response elements (GRE's). The resultant hybrid proteins activated transcription from promoters linked to the lex operator. Enhancement still required hormone binding by the hybrid receptor regardless of the exact positioning of the LexA binding domain within the protein. Thus, the unliganded hormone binding domain of the receptor acts as a strong but reversible inhibitor of receptor activity in a manner that is independent of the means by which the receptor recognizes DNA. The results also show directly that the receptor contains at least one "enhancement domain" other than that overlapping the GRE binding region; the second domain, enh2, occupies a region near the receptor amino terminus. PMID- 3043664 TI - Sexual responses are--almost--all in the brain. PMID- 3043666 TI - Characterization of a helical protein designed from first principles. AB - The question of how the primary amino acid sequence of a protein determines its three-dimensional structure is still unanswered. One approach to this problem involves the de novo design of model peptides and proteins that should adopt desired three-dimensional structures. A systematic approach was aimed at the design of a four-helix bundle protein. The gene encoding the designed protein was synthesized and the protein was expressed in Escherichia coli and purified to homogeneity. The protein was shown to be monomeric, highly helical, and very stable to denaturation by guanidine hydrochloride (GuHCl). Thus a globular protein has been designed that is capable of adopting a stable, folded structure in aqueous solution. PMID- 3043665 TI - Mammalian glucocorticoid receptor derivatives enhance transcription in yeast. AB - In mammalian cells, the glucocorticoid receptor binds specifically to glucocorticoid response element (GRE) DNA sequences and enhances transcription from linked promoters. It is shown here that derivatives of the glucocorticoid receptor also enhance transcription when expressed in yeast. Receptor-mediated enhancement in yeast was observed in fusions of GRE sequences to the yeast cytochrome c1 (CYC1) promoter; the CYC1 upstream activator sequences were not essential, since enhancement was observed in fusions of GREs to mutant CYC1 promoters retaining only the TATA region and transcription startpoints. It is concluded that the receptor operates by a common, highly conserved mechanism in yeast and mammalian cells. PMID- 3043668 TI - [Nitrous oxide anesthesia as a risk factor in arthroscopy of the knee joint using gas insufflation]. PMID- 3043667 TI - [Current immunological and clinical aspects of post-traumatic sepsis. A status report]. PMID- 3043669 TI - Long-term complications of cisplatin-based chemotherapy for testis cancer. PMID- 3043670 TI - Gonadotrophin releasing hormone analogues for prostatic cancer: an overview. PMID- 3043671 TI - Non-hormone chemotherapy for prostate cancer: principles of treatment and application to the testing of new drugs. AB - Prostate cancer, one of the commonest malignancies in western society, remains a major challenge in management. Although the typical patient is elderly and may not withstand aggressive approaches to treatment, increasing numbers of our population survive to old age, while remaining healthy and active. As prostate cancer is a disease of old age, it is therefore likely that there will be an increased requirement to combat this disease in a fit patient population. Although hormonal manipulation provides effective first-line treatment for 70% 80% of patients with metastatic disease, the majority of these ultimately relapse. Cytotoxic chemotherapy has not provided a panacea for relapsed, hormone resistant prostate cancer. Despite the significant subjective and objective responses that can be achieved by the use of single agents, the median survival of patients with hormone-resistant disease remains less than 12 months. The use of combination cytotoxic regimens has not altered this. New approaches, perhaps including the development of new cytotoxic agents, innovative uses of established drugs, or the application of the biological response modifiers will be required before this problem is resolved. Until then, we must not be satisfied with inadequate indices of success. The reporting of response rates and of survival statistics drawn from the small group of responding patients is no true indicator of success. Until truly effective treatment is available, we must learn to define more useful indicators of patient benefit, to be more effective in palliating the symptoms of this disease, and to be more critical of the limitations of our progress. PMID- 3043672 TI - Regulation of the production and function of granulocytes and monocytes. AB - The bone marrow responds to infection by rapidly producing mature granulocytes and monocytes from a small pool of committed progenitor cells under the influence of a heterogeneous family of glycoproteins termed colony-stimulating factors (CSFs). There are at least four major CSFs (IL-3, GM-, G-, and M-CSF) which are structurally distinct but have a great deal of functional overlap. The humoral signals which regulate the production of CSFs are generated at peripheral sites of infection through the activation of local macrophages and T lymphocytes by the invading pathogen. The monokines IL-1 and TNF are most likely the major humoral factors involved in stimulating CSF secretion by accessory cells in peripheral tissue sites as well as in the bone marrow microenvironment, although the functions of CSFs produced in these two organ compartments is distinct. CSFs secreted by bone marrow endothelial cells and fibroblasts serve to stimulate the proliferation and differentiation of myeloid progenitor cells, while CSFs present in areas of local infection are most likely involved in the activation of mature myeloid cell function. The dual ability of CSFs to both regulate bone marrow proliferation and to stimulate mature myeloid cell function represents a novel mechanism for producing a coordinated host response to infection. PMID- 3043673 TI - Human immunodeficiency virus: genetic structure and function. PMID- 3043674 TI - HIV infection: diagnosis and epidemiology. PMID- 3043675 TI - Hematologic manifestations of the human immune deficiency virus (HIV). PMID- 3043676 TI - Immunologic thrombocytopenic purpura in HIV-seropositive homosexuals, narcotic addicts and hemophiliacs. PMID- 3043677 TI - Infectious mononucleosis. PMID- 3043679 TI - Enteropathogenicity of E. coli. PMID- 3043678 TI - Detection of labile enterotoxin of E. coli by the Biken assay and the GM1-ELISA. PMID- 3043680 TI - Update on management of hypertension in pregnancy--medical aspects. PMID- 3043681 TI - Prenatal ultrasound estimation of foetal weight. PMID- 3043682 TI - Cardiomyopathies of infancy. AB - Cardiomyopathies in infancy constitute an ill-defined group of conditions. Recently, however, a number of specific disorders of metabolism and mitochondrial abnormalities have been described. Idiopathic or primary cardiomyopathies remain poorly defined. Endocardial fibroelastosis, the most prominent member in this group, probably represents a reaction of the endocardium to various insults. The different types of cardiomyopathy (CM) present a similar clinical picture and require the expertise of a group of specialists for proper diagnosis. Therapy is mostly supportive, as only a few cases of CM are amenable to specific treatment. Orthopic cardiac transplantation may become a therapeutic option in the future for infants unresponsive to current therapy. PMID- 3043683 TI - Parkinson's disease: recent advances in therapy. PMID- 3043685 TI - Complications of gastrostomy. AB - Despite its simplicity, gastrostomy has been associated with a relatively high incidence of complications. We have described two unusual complications: inadvertent duodenostomy, resulting in gastric outlet obstruction, and the migration of a gastrostomy feeding tube into the midportion of the jejunum, resulting in obstruction of the small bowel. PMID- 3043684 TI - Virgil Sydenstricker: special reference to niacin deficiency encephalopathy. AB - Virgil Sydenstricker was a member of a notable American family which included authoress Pearl S. Buck and the eminent epidemiologist Edgar Sydenstricker. Dr. Sydenstricker's contributions in the fields of hematology and nutritional disease are legion. His landmark work in sickle cell anemia characterized a definite symptom complex with specific hematologic findings and inheritance pattern. He wrote on the complications of malnutrition and attempted to delineate the specific effects of individual nutritional factors. Dr. Sydenstricker and his associate H. M. Cleckley first described the syndrome of niacin deficiency encephalopathy. Today, the syndrome is still occasionally reported. Niacin deficiency should be considered when unexplained acute confusional states or neurologic deficits occur in the setting of malnutrition, antituberculous drug use, or chronic partial nutritional deficiency with acute increase in metabolic demand. PMID- 3043687 TI - Blood culture positivity patterns in bacteremia due to Mycobacterium avium intracellulare. PMID- 3043686 TI - Common-source outbreak of trichinosis associated with eating raw home-butchered pork. AB - Four patients had trichinosis after consuming raw home-butchered pork. The patients had fever, myalgias, periorbital edema, and conjunctivitis. All of the patients had nausea, vomiting, or diarrhea (corresponding to the intestinal phase of the infection) seven to ten days before the onset of fever and myalgias. Laboratory findings included eosinophilia, elevated serum CPK and aldolase values, and seroconversion of Trichinella serology one month after onset of myalgias. The patients were treated with mebendazole and prednisone and recovered uneventfully. PMID- 3043689 TI - Ultrasonography in the diagnosis of appendicitis. PMID- 3043688 TI - Crohn's disease beginning during pregnancy. AB - I have reported the 11th case of Crohn's disease initially symptomatic during pregnancy. The complication rate is very high in this rare clinical syndrome. Of the ten previously described patients, five had surgical intervention during pregnancy. Seven of twelve infants and two of the 11 patients ultimately died. The prognosis is probably no better now than it was when the entity was initially described 40 years ago. PMID- 3043690 TI - Diagnostic imaging in children with urinary tract infection. PMID- 3043691 TI - Suppurative thrombophlebitis: a new look at a continuing problem. AB - Despite knowledge of its existence and despite precautionary measures, suppurative thrombophlebitis remains a significant problem in critical care, with a reported incidence of approximately 4% and associated mortality as high as 83%. In a retrospective study, we identified additional risk factors such as emergency department or field placement of catheters and administration of potassium boluses. New guidelines for surveillance and prevention have proved successful. PMID- 3043692 TI - Treatment of otitis media with cefuroxime axetil. AB - Cefuroxime axetil and cefaclor were compared for efficacy in the treatment of acute otitis media with effusion. Sixty-four pediatric outpatients had tympanocentesis for culture, and then were randomized to a ten-day course of treatment with cefuroxime axetil or cefaclor. Streptococcus pneumoniae and Haemophilus influenzae were isolated from 25 (39%) and 23 (36%) patients, respectively. Treatment was beneficial in 26 (90%) of the patients who received cefuroxime axetil, and in 16 (76%) of the cefaclor-treated patients. Treatment failed in five (24%) of the cefaclor-treated patients, and in only three (10%) patients who received cefuroxime axetil. Haemophilus influenzae was the initial causative pathogen in a disproportionate number of treatment failures. This study demonstrates the efficacy of cefuroxime axetil in the treatment of otitis media. PMID- 3043693 TI - Influence of cross-education training in postoperative hand therapy. AB - Early studies have shown an effect of cross-education training in normal volunteers and in patients with neuromuscular disorders. In this prospective study, I evaluated application of this principle after upper extremity surgery and immobilization. A detailed therapy program to the contralateral unoperated extremity was initiated in one half of the patients. The other half of the patients had only routine postoperative instructions for rest, elevation, and protective splinting. Range of motion and strength measurements were done preoperatively and at intervals up to three months after operation. Analysis of results shows little difference in range of motion measurements in either group. However, return of strength in the extremity operated on was significantly augmented, up to 150% of the control group, in the patients who received cross education training. Application of this principle to rehabilitation after injury to specific functional units may increase recovery. PMID- 3043695 TI - Applications of biotechnological methods to studies of protozoan parasites. AB - Biotechnology applies and extends the concepts and techniques of molecular biology. An overview of the applications and potential uses of the technology is presented for selected protozoan parasites. The areas reviewed include the characterization of protozoa, the production of their antigens, and the uses of hybridomas for studies of the antigens and host responses. In addition to the traditional methods of classification, parasitic protozoa are identified and characterized according to stable molecular markers. Peptidemes, zymodemes, antigens, schizodemes, and chromosomal complements define isolates and correlate with biological activities of the parasites. While antigens are typically extracted from parasites obtained from infected hosts or grown in vitro, they may be produced with in vitro translation systems, recombinant procedures with bacteria, or more recently developed techniques such as co-transformation and electroporation with eucaryotic cells. Monoclonal antibodies produced by B-cell hybridomas are used to identify parasites, antigens, epitopes, and the locations and functions of specific antigens. T-cell hybridomas may provide insights into cell-mediated immunity and interactions with the parasites. PMID- 3043694 TI - Intrathoracic lymphadenopathy in postprimary tuberculosis. AB - In the past, hilar or mediastinal lymphadenopathy was considered by many to be a feature of only the primary or first infection with Mycobacterium tuberculosis, and to exclude the diagnosis of reactivation or postprimary tuberculosis. In a series of 56 adult patients with documented postprimary disease due to M tuberculosis, we found hilar or mediastinal lymphadenopathy in three cases (5%). Although intrathoracic lymphadenopathy was more common in primary tuberculosis, we do not believe that intrathoracic lymphadenopathy is as specific for primary tuberculosis, particularly in the adult, as was once thought. For this reason, we believe that the roentgenographic demonstration of intrathoracic lymphadenopathy should not be used as a definitive research or clinical criterion for primary tuberculosis in an adult. PMID- 3043696 TI - Immunodiagnosis of clonorchiasis by enzyme-linked immunosorbent assay. AB - The present study applied the indirect enzyme-linked immunosorbent assay (ELISA) technique in the immunodiagnosis of clonorchiasis. Antigen used in this study was extracted from adult worms of Clonorchis sinensis obtained from cats. 132 patients with clonorchiasis, 100 healthy persons and 14 patients with other parasitic infections were studied. Mean O.D. ratio with standard deviation of clonorchiasis was 1.41 +/- 0.21 with 0.95 +/- 0.13 of healthy persons. Results revealed 90.2% to 95.5% of sensitivity and 84% to 99% specificity dependent on the two cut off values of O.D. ratio, i.e. 1.10 and 1.20. Antibody titers derived from O.D. ratio highly correlated with direct titration (Y = 0.0303 +/- 1.1766 X, r = 0.8945). Cross reactions of other parasite infections to clonorchiasis were observed in patients with angiostrongyliasis and schistosomiasis. PMID- 3043697 TI - Nematode isoenzyme studies and cytogenetics. AB - The current information on isoenzyme studies of nematode parasites was reviewed. The genetic heterogeneity as reviewed by these studies was highlighted. Application of isoenzyme studies and the role of biotechnological techniques in isoenzyme studies was discussed, and the status of cytogenetic studies on nematode parasites was presented. PMID- 3043698 TI - Development of immunoradiometric assay for detection of Plasmodium falciparum antigen in blood using monoclonal antibody. AB - Specific monoclonal antibody (MAB) F2W22C1) produced by hybridoma technology against blood stage common antigens shared by almost all isolates of P. falciparum was selected. The 125I-labelled prepared from this MAB (125I-MIgG) was used as probe for detection of low grade P. falciparum infection. Recently, a two site monoclonal antibody sandwich immunoradiometric assay (Mab-IRMA) has been developed. The assay showed good correlation with parasitaemia with the ability to detect as few as 2.4 parasites/10(8) erythrocytes. Two surveys were made in people living in a malaria endemic area during transmission and non-transmission season to detect the antigen of Plasmodium falciparum by using the Mab-IRMA. In the first survey involving 101 people, the IRMA positive rate was 56.4% and then significantly declined to 16.5% during the second survey involving 79 people of the same group. The parasitological positive rates were likewise decreased from 11.9% to 1.3% (p = 0.015) during these two seasons. IRMA positive rates were significantly higher than the corresponding parasitological positive rates (p less than 0.0001 and less than 0.002 for the first and the second surveys respectively). Regression analysis showed that IRMA activities were linearly correlated with the parasite counts by microscopic examinations (r = 0.629, p = 0.022). IRMA was specific for P. falciparum since all 30 healthy controls and six of seven vivax malaria cases were negative. IRMA has potential for use to monitor the malaria control and to assess the efficacy of the future field trial of malaria vaccine. PMID- 3043699 TI - Biotechnological methods on the study of nematodes. AB - During the past several years, a great many advances have been made in obtaining a better understanding of the biology of nematode parasites. Much of the information obtained has been with the use of tools developed in the expanding field of biotechnology, or more specifically, the use of biochemistry, genetics or molecular biology. Advances are being made on speciation of parasites, their function and pathogenesis. Some nematode parasites indigenous to Southeast Asia have been studied, but much more should be done. There is a large group of parasites in Southeast Asia that could be studied. In the past, emphasis has been on human nematodes, but there are many interesting parasites of lower animals that should be investigated. Furthermore, future parasitologists must first be trained in basic parasitology and then be exposed to new technology in order to apply these tools to further our knowledge of round worms. PMID- 3043700 TI - Application of biotechnology methods to the study of cestodes. AB - The advent of biotechnology has invigorated research on the control of cestode diseases, especially cysticercosis infections in man and animals. The utilization of hybridoma technology to produce antigen-specific monoclonal antibodies has resulted in great strides towards obtaining pure antigens relevant for immunodiagnostic purposes and for research on vaccines. However, the isolation and identification of antigens is only the initial step in the development of such reagents. Production of antigens in quantities sufficient for research/development and commercialization is hampered by the scarcity of viable parasite material for extraction. Expectations are that this problem can be surmounted by application of recombinant DNA methods to produce cloned genes for antigen expression in cultured microorganisms or cells. Remarkable progress has been made recently in isolating and cloning genes from several important cestode species and antigens have been expressed in vitro with genes cloned from Taenia taeniaformis and T. hydatigena. Although these early efforts have not as yet resulted in practical antigen production, the prospects for doing so appear good. The complex epidemiology of cestode diseases is another research subject that has benefited from the successful application of the tools of biotechnology. For example, the greater precision in typing biological variants afforded by DNA analysis has led to important revisions of the understanding of hydatid disease. DNA probes are now available for Echinococcus spp, which are effective for typing isolates. These probes may also find use as reagents for distinguishing eggs of Echinococcus from other taeniid eggs, a serious difficulty for field investigations.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3043702 TI - Application of biotechnology to the study of molluscan hosts of animal parasites. AB - The intimate and complex relationship that exists between man and mollusc makes the group an ideal target for the application of biotechnology. The paper discusses and reviews the application of both the conventional and new molecular techniques to the study of the molluscan hosts of animal parasites. The remarkable advances in molluscan biology which have occurred during the past two decades as a consequence of conventional technological methods are noted and applications pertinent to problems in Southeast Asia are cited. Applications of new biotechniques, including those of molecular biology, to problems of the host parasite interaction are discussed. Examples of how the new biotechnology may resolve present and future problems in applied malacology are presented. PMID- 3043701 TI - Biotechnology research on cestodes in the Philippines, Malaysia, and Indonesia. AB - There are essentially no reports on the use of modern biotechnological methods on the study of cestode parasites in the Philippines, Indonesia or Malaysia. The only recent reports of cestode studies in these countries have been on reports of new species in animals and on prevalence rates of cestode parasites in humans; Taenia solium and cysticercosis, Taenia saginata and Hymenolepis nana, etc. Reports on the use of biotechnology has emanated from outside the area on cestodes of humans and animals, and some of these methods could be used to study cestodes in this part of the world. PMID- 3043703 TI - Application of biotechnology methods to the study of trematodes. AB - Advances in biotechnology over the past decade offer renewed hope for solutions to trematode parasitoses in Thailand. Trematode diseases have remained the same, but the tools (1) to exploit the innate ability of cells to replicate and produce biological products upon demand, (2) to manipulate the genetic makeup of an organism, (3) and to biologically or synthetically manufacture peptides have provided scientists with new reagents for diagnosing, treating, preventing and controlling trematode diseases. Although recent applications have been focused on schistosomiasis, they have potential application to other trematode diseases (paragonimiasis, clonorchiasis, opisthorchiasis, fasciolopsiasis, and fascioliasis) endemic to Thailand. The optimism which new technology has generated must be tempered with realism and continued support for basic parasitology. Progress is dependent on a combination of approaches involving techniques at the molecular, cellular, organism and population levels. The new tools must be used in conjunction with old knowledge. Most of all there must be a rational strategy for the use of products of new technology. Defined vaccines alone will not be the answer, even if they become a reality. They must be applied in a manner that optimizes their effectiveness. The silver bullets may be out there, but they are still in a roughly molded stage. They must be finished, loaded in an appropriate epidemiological gun, and fired by an expert at specific targets. PMID- 3043704 TI - Adaptation of biotechnology methods on the study of arthropods. AB - Biotechnological advances are rapidly being applied to the study of arthropods and arthropod-born diseases. Improved tools are available for species identification, study of species complexes, detection and identification of pathogens in vectors and detection and characterization of insecticide resistance. These techniques have their basis in hybridoma, DNA probe and biochemical assay technologies. Problem areas in assay development are discussed. PMID- 3043706 TI - [Methods of examining the biliary tract in acute cholecystitis in elderly patients]. PMID- 3043705 TI - Investigation of malaria sporozoites by immunoradiometric assay. AB - Two-site immunoradiometric assay (IRMA) (Zavala et al., 1982) using monoclonal antibodies to P. falciparum and P. vivax was applied to detect sporozoites in laboratory-maintained An. dirus and also mosquitoes collected from endemic areas of malaria in Thailand. Study in P. falciparum infected mosquitoes revealed that the circumsporozoite (CS) antigen was first found in the abdominal portion on day 10 post-infection, while it could be observed in the salivary glands from day 15 onwards. The head-thorax portion of wild-caught mosquitoes were investigated by IRMA compared with the dissection technique. The results showed that none of the mosquitoes collected from Phrae was positive for malaria. The mosquitoes collected from Chantaburi showed 4 out of 1243 An. dirus that were positive for P. falciparum by IRMA, with sporozoites ranging from 207 to 3875. Among 3123 An. minimus collected from Kanchanaburi, 3 were positive by IRMA, 2 for P. falciparum and one P. vivax with sporozoites found in head-thorax portion were 1880, 2380 and 1026 respectively. Not a single sporozoite was found in the mosquitoes collected from these areas by the dissection technique. However 7 out of 1219 An. minimus from Kanchanaburi were found to possess undeveloped oocysts in the stomach wall. It is evident that the IRMA is efficient, convenient and suitable for the investigation of sporozoites in this region. The application of this technique in further epidemiological study of malaria is in progress. PMID- 3043708 TI - [I.A. Kassirskii and his contribution to Soviet medicine (on his 90th birthday)]. PMID- 3043707 TI - [Status of lipid metabolism in diabetes mellitus type 1 depending upon the degree of compensation of carbohydrate metabolism]. PMID- 3043709 TI - [Insulin as a regulator of hydrogen ion homeostasis]. PMID- 3043710 TI - [Status of the prostacyclin-thromboxane A2 system in patients with ischemic heart disease during induced myocardial ischemia]. PMID- 3043711 TI - [Restoration of intestinal continuity in patients with colostomy]. PMID- 3043712 TI - [Primary plastic surgery of flexor tendons and nerves in their combined injuries in proximal segments of the fingers]. PMID- 3043713 TI - [Hemodynamics of the lesser circulation in lung diseases]. PMID- 3043714 TI - [Physical training as a method of non-pharmacological therapy of hypertension]. PMID- 3043715 TI - [Pharmacodynamics of cordipine in hypertension]. PMID- 3043716 TI - [The renin-angiotensin-aldosterone system in chronic diseases of the liver]. PMID- 3043717 TI - Quantitative assessment of human proteinases as agents for chemonucleolysis. AB - A rabbit model system is described. It allows accurate measurement of the dose dependent loss of glycosaminoglycan from the nucleus pulposus of lumbar intervertebral discs after injection of a proteinase. At the dose equivalent to that of chymopapain used in human chemonucleolysis, two human serine proteinases, cathepsin G and chymotrypsin, were as effective as chymopapain in removing up to 80% of the glycosaminoglycan from the disc. A cysteine proteinase, cathepsin B released no more than 45% of glycosaminoglycan. X-ray films clearly showed narrowing of the disc space when 30-40% of glycosaminoglycan was removed. The degradation of the nucleus pulposus was seen histologically as loss of toluidine blue metachromasia. PMID- 3043718 TI - Histologic and morphometric observations on vertebral bone of aging sand rats. AB - The trabecular bone of the vertebrae of 30 male and 30 female sand rats (Psammomys obesus) aged 13 to 33 months was examined histologically and morphometrically. The usual age-linked decline of bone mass failed to occur in females and was statistically not significant in males. A few changes with age were noted at the cellular level. Sex differences were statistically significant only in animals living into the third year of life, males having a smaller bone mass than females. This abnormality of the vertebral spongiosa is attributed to pathologic local stresses caused by the numerous instances of disc degeneration and herniation. Differences in size, location, and age of the herniations are thought to account for the wide fluctuations in the individual bone parameters examined. Whereas the spine of the sand rat provides an excellent model for the study of spondylosis, it is unsuited as a model for age-linked osteoporosis. PMID- 3043720 TI - Spinal cord compression: an exceptional complication of spinal osteoporosis. PMID- 3043721 TI - Proteus infection of the spine. PMID- 3043719 TI - Laminar removal and replacement: a technique for the removal of epidural tumor. PMID- 3043722 TI - [Diagnosis of placenta praevia using ultrasound]. PMID- 3043723 TI - [Arthroscopic surgery]. PMID- 3043724 TI - Diabetes mellitus, hypertension and insulin. PMID- 3043725 TI - An introduction to clinical epidemiology. AB - Clinical epidemiology is defined and its influence on methods of diagnosing and treating patients assessed. A plea is made for the wider application of such methods. PMID- 3043726 TI - Pseudoxanthoma elasticum in South African black patients. A report on 7 cases. AB - Pseudoxanthoma elasticum in 7 South African black patients is reported. All demonstrated cutaneous lesions of varying severity and extent. Gastro-intestinal bleeding requiring surgical intervention was found in 1 patient. PMID- 3043727 TI - Pancreatic transplantation and ductovesical drainage. PMID- 3043729 TI - The petroleum industry. PMID- 3043730 TI - Occupational medicine in the petroleum industry: an historical perspective. AB - The development of occupational health in the petroleum industry has been reviewed from the early years of the 20th century through the 1970s. Approximately 50 years after its chaotic beginning, the industry began to realize its obligations to its employees and undertook activities that were already standard practice in a number of other business enterprises. After a belated start and slow progress for several decades, by mid-century practically all of the major oil companies were involved in the principal activities of modern occupational health. Significant advances had been made in applying preventive measures in a variety of environments throughout the world. Over the following 30 years the level of clinical practices and the associated professional disciplines of industrial hygiene, toxicology and epidemiology demonstrated the industry's leadership to its peers as well as to society in general. PMID- 3043731 TI - Occupational medicine in the eighties: decade of change. AB - This chapter focuses on current medical practices in the petroleum industry as they reflect the social, managerial, technological and regulatory changes of the present decade. Modern health policies and medical organizations are reviewed, with emphasis on health evaluation programs, health information distribution and clinical services. The author concludes with commentary about apparent future trends. PMID- 3043728 TI - Actraphane and Lentard insulin for once daily insulin supplementation. PMID- 3043732 TI - Petroleum: its composition, analysis and processing. AB - The task of the occupational physician can be greatly assisted by a knowledge of the manufacturing processes in which workers are engaged, the nature of the products that these processes produce, and the raw materials upon which they depend. Because this is particularly true of the petroleum industry, this chapter is devoted to a challenging overview of petroleum composition, analysis and refining technology. PMID- 3043733 TI - Occupational exposures to potentially hazardous agents in the petroleum industry. AB - This chapter has been created to acquaint the reader with occupational exposures that are more common in, and somewhat unique to, the petroleum industry. Both highly toxic materials capable of causing acute illness or even death following short-term exposure, and chemical and physical agents that pose risk of chronic and irreversible damage to health during prolonged exposure are addressed. PMID- 3043734 TI - Toxicology of petroleum hydrocarbons. AB - Before discussing the toxicologic effects of petroleum hydrocarbons, the author reviews the considerable resources available today to investigators in this area. He then covers the toxicology of the major classes of petroleum hydrocarbons, including aliphatic, alicyclic and aromatic compounds. PMID- 3043735 TI - Epidemiologic studies of the petroleum industry. AB - This paper reviews the epidemiologic studies of petroleum workers published in the English language, focusing on research pertaining to the petroleum industry, rather than the broader petrochemical industry. Types of epidemiologic studies are described, then overall cause-specific mortality and morbidity among workers. PMID- 3043736 TI - The carcinogenic potential of selected petroleum-derived products. AB - In this chapter the authors examine the toxicologic and epidemiologic literature for a broad range of petroleum-derived products in order to assess the carcinogenic potential of these compounds. Types of evidence used, classes of compounds, and qualitative assessments of carcinogenicity are presented. PMID- 3043737 TI - A review of the non-neoplastic kidney effects of hydrocarbon exposure in humans. AB - This review has demonstrated that there is a considerable amount of information in the medical literature concerning hydrocarbon-associated kidney effects. The existing data lends itself to a variety of divergent interpretations. Ravnskov has stated that "glomerulonephritis should be recognized legally as an occupational disease," yet there is no mention of hydrocarbon exposure in the differential diagnosis of glomerulonephritis in two standard American textbooks of internal medicine. Two recently published textbooks of occupational medicine state without reservation that "studies have linked hydrocarbon exposure to glomerulonephritis" and base this conclusion on the previously cited studies of Beirne and Brennan, Zimmerman, and Ravnskov. Based on this review, the following conclusions have been reasonably substantiated: 1. Massive exposure to petroleum distillates on rare occasions may cause acute renal failure due to tubular necrosis. This appears to be a reversible lesion which, depending on the level of exposure, the medical care and support available, and pre-existing renal function, may be without chronic sequelae. 2. Case reports linking Goodpasture's syndrome and other types of glomerulonephritis to hydrocarbon exposure are based on circumstantial evidence and cannot be used to establish a causal association. 3. The evidence from the eight case-control studies of hydrocarbon exposure and glomerulonephritis is inconclusive. Six of the eight published case-control studies show a positive association between hydrocarbon exposure and glomerulonephritis, but four of the six studies have methodologic flaws that could explain the observed effect. The findings in the one positive study that is methodologically acceptable were not replicated in a subsequent study utilizing a similar design. 4. Studies of hydrocarbon-exposed occupational cohorts have generally revealed a lower than expected risk of death from renal causes. As with most historical cohorts, the specific exposures, intensities and durations of exposure have been poorly defined. Effects of mortality that may occur among highly exposed subsets of these occupational cohorts may be diluted by a relatively large proportion of workers with minimal or no exposure to the class of hydrocarbons in question. 5. Studies of renal biochemistry and renal function effects have been uniformly negative in groups of workers from several industries with relatively high exposures of long duration to a variety of hydrocarbon solvents. The statistically significant differences in proteinuria and cell excretion observed in one of the studies should not be confused with clinical significance.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3043738 TI - Benzene toxicity. AB - The proper control of benzene in the workplace and the general environment has been a subject of relatively intense interest for at least the past decade. The author reviews the toxicology of benzene, its hematologic effects in the human, and methods of measurement of benzene exposure in the workplace. PMID- 3043739 TI - Health-related research at the American Petroleum Institute. AB - On behalf of its oil-company members, API has studied the effects of the materials to which people may be exposed and aspects of the effects of the total work environment on employees' health. This chapter sets out the mechanisms by which the research is conducted, illustrates the type of research performed, and describes the availability of the results to those interested in the scientific community. PMID- 3043740 TI - Posterior scleritis. AB - Posterior scleritis must be considered in the differential diagnosis of many ocular conditions, including angle closure glaucoma, choroidal folds, optic disk edema, circumscribed fundus mass, choroidal detachment, and exudative retinal detachment. Because it is rare, a high index of suspicion is necessary. Anterior scleritis, pain, or a history of collagen-vascular disease, when present, help to alert the clinician to the correct diagnosis. Posterior scleritis affects women more often than men, but annular ciliochoroidal effusion and choroidal folds are more common in men. Exudative macular detachment and a circumscribed fundus mass are more common in women. This paper reviews the world literature on posterior scleritis and describes findings in a series of 43 patients seen at Wills Eye Hospital. It stresses the clinical features and ancillary diagnostic tests that help to establish the diagnosis. PMID- 3043741 TI - Galactose and cataract. AB - Galactosemia is a disorder caused by a deficiency of any one of three possible enzymes involved in the metabolism of galactose: galactokinase, transferase or epimerase. Any single deficient enzyme can result in cataract through the accumulation of galactitol in the lens. The ophthalmologist may play an important role in this disease, since early recognition of cataract development followed by the initiation of a galactose-free diet may lead to clearing of lenticular opacities. The clinical and laboratory findings that distinguish the three enzyme deficiency disorders of galactosemia are discussed. The biochemical genetics of each enzyme also are reviewed, along with the recent evidence linking heterozygous galactokinase deficiency and presenile cataract. PMID- 3043744 TI - History of drug abuse treatment in Texas. PMID- 3043742 TI - Radiography of the eye and orbit: a historical overview. AB - The history of radiography and orbital imaging begins in 1895 with Wilhelm Roentgen's discovery of x-rays. Over the next three quarters of a century, radiographic pioneers like Dr. William Sweet, who developed the Sweet method, and Dr. George E. Pfahler, who made the first successful pictures of a brain tumor, helped to bring radiography into the 20th century. With each new radiologic innovation producing a forward surge followed by a period of refinement, new methods were invented and utilized to their diagnostic limits. But perhaps none of the radiologic innovations of this century--the Coolidge tube, the Potter Bucky diaphragm, tomography and angiography-will have more impact than computed tomography and magnetic resonance imaging. PMID- 3043743 TI - Absele in bone hemostasis--a clinical and experimental investigation. AB - Control of bleeding from bone tissue may sometimes be difficult to maintain. Existing methods of haemorrhage control with standard bone wax can interfere with subsequent healing. This paper presents clinical and experimental results of the use of a new absorbable bone sealant, Absele a paste made from stabilised fibrin and soluble collagen. PMID- 3043745 TI - [Clearance mechanisms of the airways]. PMID- 3043747 TI - [An appraisal of bronchial cancer]. PMID- 3043746 TI - [Auscultation of the lung]. PMID- 3043748 TI - [Pleural effusion of unclear origin: practical diagnostic procedure]. PMID- 3043749 TI - [Diagnosis of lung emphysema: conventional methods and computerized tomography]. PMID- 3043750 TI - [Inhalation therapy]. PMID- 3043751 TI - [Influenza and pneumococcal vaccination in patients with chronic obstructive lung diseases: uses and risks]. PMID- 3043752 TI - [High altitude pulmonary edema: epidemiology, clinical aspects, pathogenesis and therapy]. PMID- 3043753 TI - Pulmonary function after bone marrow transplantation for chronic myeloid leukaemia. AB - Pulmonary function was measured before and at intervals after treatment in 44 patients who received a bone marrow transplant for chronic myeloid leukaemia in the chronic phase. All patients were treated with cytotoxic drugs, total body irradiation, and post-graft immunosuppression. Thirty four patients surviving for 12 months were followed at three monthly intervals and 16 patients for 24 months. Fifteen patients received unmanipulated donor marrow cells and 29 patients received donor marrow cells depleted of lymphocytes ex vivo with the monoclonal antibody Campath-1. The 21 patients treated early in this study received 10 Gy of total body irradiation whereas the 23 patients treated more recently, who were all T lymphocyte depleted, received 12 Gy. Pretransplant lung function for the group was normal and was similar in survivors (n = 34) and nonsurvivors (n = 10), and in smokers (n = 8) and non-smokers (n = 36). (Carbon monoxide transfer factor -TLCO) was under 75% of predicted normal in nine patients before transplantation. TLCO, carbon monoxide transfer coefficient (KCO), FEV1, and vital capacity (VC) values were lower 6 and 12 months after bone marrow transplant than initially. The greatest decline was in TLCO, from an initial value of 89% to 66% at 6 and 70% at 12 months. The 16 longer term survivors showed significant recovery of function between 6 and 24 months after bone marrow transplant for TLCO, KCO, and VC, the increase ranging from 6.3% to 7.3% predicted. Airflow obstruction (FEV1/VC ratio less than 70%) developed in one patient. The major factors associated with deterioration in pulmonary function at 6 and 12 months after transplantation in the 34 survivors (stepwise multiple regression analysis) were (a) transplantation with T cell depleted donor marrow (p less than 0.005) and higher total body irradiation dose (p less than 0.02) with a fall in KCO and an increase in the FEV1/VC ratio; (b) chronic graft versus host disease with a fall in VC (p less than 0.01); and less fall in KCO (p less than 0.01); and (c) acute graft versus host disease with a fall in FEV1 (p less than 0.01). It is considered that most patients who survive the short term risks of bone marrow transplant have only minor long term impairment of pulmonary function. PMID- 3043755 TI - [Drug-induced hemolytic anemia. Results of consensus conferences]. PMID- 3043754 TI - Intrathoracic infections due to Eikenella corrodens. PMID- 3043756 TI - [Resistance to amphotericin B and genotypic resistance to 5-fluorocytosine of serotypes A and B of Candida albicans]. PMID- 3043757 TI - [Effects of alprazolam on arousal, memory and induced stress in the healthy volunteer]. PMID- 3043758 TI - [Evaluation of the antihypertensive effect of delayed-action nicardipine with 24 hour ambulatory blood pressure monitoring]. PMID- 3043759 TI - Expression of polymorphic B-cell antigens on human kidneys. AB - We have examined the expression on a panel of 22 human kidneys of polymorphic B cell determinants recognized by mouse monoclonal antibodies. Monoclonal antibodies from a mouse immunized with an antigenic preparation from a DR4 positive B-cell line reacted preferentially with kidneys from DR4 positive donors (p less than 0.005), and the pattern of reactivity with kidney tissues was similar to that of antibodies to monomorphic determinants of class II. However, these antibodies did not show clear specificity for DR4 on lymphocytes in standard serological analyses. These results provide evidence for the expression of polymorphic class II determinants on human kidneys. Reasons for the differences in the apparent specificities of the monoclonal antibodies when tested on kidney sections and lymphocytes are discussed. PMID- 3043760 TI - HLA-DR expression by adrenocortical cells of the zona reticularis: structural and allotypic characterization. AB - We previously reported that human adrenocortical cells in the zona reticularis of normal glands express antigenic determinants recognized by HLA-DR monoclonal antibodies (MoAbs). In the present study, it is shown that these adrenocortical HLA-DR determinants are present on glycoproteins having similar structural characteristics, regarding subunit composition and molecular weight, to those of HLA-DR molecules present on immunocomponent cells. Furthermore, adrenocortical HLA-DR molecules include serologically-defined genetically-appropriate allotypic specificities, detectable by immunostainings with both HLA-DR human alloantisera and MoAbs against polymorphic HLA-DR determinants. The finding of a normal expression of HLA-DR antigens by these highly differentiated and biosynthetically active endocrine cells gives support to the notion that MHC class II molecules may perform biological functions in addition to those related to the immune response. PMID- 3043761 TI - Morphometric analysis of Rhodnius prolixus Stal (Hemiptera:Reduviidae) midgut cells during blood digestion. AB - Post-feeding ultrastructural modifications to the midgut cells of Rhodnius prolixus are quantified using morphometry. Changes in relative and absolute volumes and/or surface areas are demonstrated for the whole cells, nuclei, mitochondria, rough endoplasmic reticulum, lysosomes, Golgi apparatus, storage vesicles, glycogen, microvilli, and basal labyrinth, before and during blood digestion. These parameters are separately determined for cells from each of the three midgut regions, and are correlated against previously published cycles of digestive enzyme activities. The results support the proposed division of the midgut of R. prolixus into three functional regions: the anterior midgut or crop is the site of water transport immediately after feeding, and of lipid and glycogen storage. No protein digestion occurs in this region. The anterior intestine is the site of most proteinase synthesis and secretion, although limited absorption and nutrient storage also occurs. The posterior intestine is responsible for some secretory activity, but is also implicated as the most important region for absorption of digested nutrients and for carbohydrate absorption and storage. PMID- 3043764 TI - Induced abortion in sub-Saharan Africa: what we do and do not know. AB - The first step in addressing the growing public health problem of abortion in sub Saharan Africa is to gain a better understanding of the problem and its complexities. Abortion behavior is inextricably connected with issues of women's roles and opportunities, and until the various dimensions of abortion behavior and its socioeconomic context are understood, governments will have difficulty addressing the problem effectively. In addition, abortion needs to be studied within the broader framework of reproductive health. In a continent where fertility is highly valued and infertility prevalent, the interaction between abortion, practice of contraception, and fears of infertility must be fully understood if we are to have any hope of reducing the numbers of unwanted pregnancies and the morbidity and mortality caused by induced abortion. PMID- 3043766 TI - Biogenesis and intracellular transport of intestinal brush border membrane hydrolases. Use of antibody probes and tissue culture. PMID- 3043762 TI - Effect of certain trace elements on the mutagenicity of N-methyl-N'-nitro-N nitrosoguanidine. AB - Tests have been carried out to detect inhibitory activity of various trace elements on mutagenesis induced by the carcinogen N-methyl-N'-nitro-N nitrosoguanidine in Salmonella typhimurium strain TA100. Selenium has been found to be most active in this regard while copper has displayed moderate inhibitory ability. The action of selenium is mediated through an interaction resulting in rapid deactivation of the carcinogen. PMID- 3043763 TI - The effects of improved child survival on family planning practice and fertility. AB - The relationship between improvements in child survival, family planning, and fertility is viewed here as the outcome of a process of family building that evolves through distinct phases as the mortality transition progresses. The speed with which family building strategies evolve from "family building by fate" to "family building by design" and from "insurance" to "replacement" as child survival improves depends on the pattern (by age and causes of death) of mortality decline and the sociocultural context. While child survival improvements will not lead to compensatory declines in fertility when fate or replacement behavior govern family building, more than compensatory fertility declines can result when families shift to family building by design, which, in its initial phases, is manifested by so-called insurance behavior. A literature review supports these hypotheses and identifies family planning availability as a critical additional factor. These results provide strong support for an integrated approach to the delivery of health and family planning services. PMID- 3043765 TI - Expression of the ABH, Lewis, and related antigens on the glycoproteins of the human jejunal brush border. PMID- 3043767 TI - Immunoaffinity purification of membrane fractions from mammalian cells. PMID- 3043768 TI - The use of antibodies for analysis of the secretory and endocytic paths of eukaryotic cells. PMID- 3043769 TI - Monoclonal antibodies in investigations on astrocytes. PMID- 3043770 TI - Current molecular approaches to experimental thyroid autoimmunity. PMID- 3043771 TI - The immunochemistry of some blood group antigens. Relation to erythrocyte membrane structure and to hemagglutination. PMID- 3043773 TI - Nightingale's environmental theory--a 20th century reality. PMID- 3043772 TI - Biochemistry and pathophysiology of the molecular forms of cholinesterases. PMID- 3043774 TI - Sinusoidal lining cell damage: the critical injury in cold preservation of liver allografts in the rat. AB - We have previously defined viability limits in a rat transplantation model. All liver allografts stored in a simple preservation solution (NaCl 0.9%, CaCl2 2 mM) at 4 degrees C for 4 hr or at 37 degrees C for 1 hr were viable upon transplantation, but all those stored at 4 degrees C for 8 hr or at 37 degrees C for 2 hr were nonviable. Only cold-preserved, nonviable livers showed increased vascular resistance, platelet trapping and an initially low, but then high, rise in aspartate transaminase (AST) upon reperfusion, all suggesting injury to the microcirculation, with secondary injury to the hepatocyte. In the present study, we investigated the morphological changes that occur in livers stored for the defined critical times, using light and electron microscopy after perfusion fixation. Accurate and reproducible identification of specimens as belonging to viable or nonviable and warm- or cold-preserved could be made in this way. Preservation in the cold first resulted in reversible changes consisting of cellular swelling, alterations of intracellular organelles, and partial denudation of the sinusoidal lining (cold-preserved viable group). Later, under conditions of nonviable cold preservation, detachment of cell bodies of sinusoidal lining cells with nuclear changes and almost complete denudation of the sinusoidal lining was observed. Endothelial cells of larger vessels were only injured mildly. In contrast, under conditions of warm preservation, changes involving mitochondria and later nuclei were found in hepatocytes, and blebbing was more extensive. Endothelial cells were spared relatively. We also examined livers stored in isotonic citrate solution at 4 degrees C for 8 hr and 16 hr, the critical times determined for this solution in another model of rat liver transplantation. The findings were very similar to storage in saline with respect to the changes in the sinusoidal lining cells after cold preservation for the two critical times. The results provide convincing evidence of a qualitative difference between warm and cold preservation injury, with relatively selective damage to hepatocytes or sinusoidal lining cells, respectively. Endothelial damage represents the primary event, resulting in the loss of organ viability following hypothermic storage. Thus morphology may serve as a useful viability marker after preservation. PMID- 3043775 TI - Successful 72-hour cold storage of dog kidneys with UW solution. AB - Effects of three cold-storage solutions on kidney function in dogs were examined with the isolated perfused (IPK) kidney model and the autotransplant model. EuroCollins' (EC) solution, phosphate-buffered sucrose solution, and a new solution developed at the University of Wisconsin (UW) were studied. Kidneys were cold-stored for 48 hr or 72 hr. With the IPK model, cold storage for 48 hr or 72 hr in each of the three solutions caused creatinine clearance to decrease by 80% 90%. More protein was excreted by kidneys stored for 48 hr in PBS solution than by kidneys stored in EC or UW solution; protein excretion after 72 hr of storage was similar for kidneys stored in EC or UW solution. Sodium reabsorption decreased after 48 hr or 72 hr of storage, but was higher in kidneys stored in UW solution (83% and 56%, respectively) than in EC solution (52% and 22%, respectively). With the autotransplant model, 40% of the kidneys were viable after 48-hr storage in PBS solution, but 80% viable when stored in EC solution and 100% were viable when stored in UW solution. All kidneys were viable when stored for 72 hr in UW solution; none were viable when stored for 72 hr in EC solution. These results suggest that UW solution effectively preserves kidneys for 72 hr. We previously reported successful 72-hr pancreas preservation. Recently UW solution was able to preserve canine livers for 30 hr. Thus, this single solution appears to be effective for preserving all intraabdominal organs and may simplify cold storage of organs for transplantation. PMID- 3043776 TI - Experimental cardiac allograft survival across major histocompatibility complex barriers in the rhesus monkey following T lymphocyte-depleted autologous marrow transplantation. I. In vitro T lymphocyte depletion studies. AB - We have developed a rhesus monkey model consisting of myeloablative total-body irradiation and T lymphocyte-depleted autologous bone marrow transplantation followed by MHC-mismatched heterotopic cardiac allograft implantation that has provided an opportunity to study the role of marrow T cells in cardiac allograft rejection. In order to assess quantitatively the effects of low numbers of residual marrow T cells following depletion, methods to deplete rhesus marrow extensively and to detect residual T cells following depletion at levels below the sensitivity of standard assays have been developed. A rhesus marrow limiting dilution assay has been developed that quantifies less than 1 T cell in 10(5) marrow cells and is superior to traditional detection methods by at least 3 logs. In a direct comparison of four T cell depletion methods, effective depletion has been achieved with complement-mediated cytotoxicity (C'MC), erythrocyte rosetting, and counterflow centrifugal elutriation (CCE), the latter with a simplified single-flow rate protocol. Median marrow T cell depletions of 2.1, 1.1, and 3.1 logs, and total nucleated cell losses of 40%, 61%, and 42% respectively, have been observed. A reported use of ricin A-chain-like toxins for the enhancement of C'MC was of low efficacy with rhesus peripheral blood T cell targets. CCE followed by C'MC has resulted in a median 4.8 logs depletion with residual marrow T cell contents less than 0.001%. Thus, C'MC, E-rosetting, and particularly CCE are effective methods of T cell depletion--and, when used in combination, extensively eliminate marrow T cells. A rhesus marrow limiting dilution assay detects residual T cells at these low levels. These techniques provide a basis for the quantitative study of the role of T cells in organ graft rejection following T lymphocyte-depleted autologous marrow transplantation. PMID- 3043777 TI - Recipient preparation for bone marrow transplantation. I. Efficacy of total-body irradiation and busulfan. AB - The efficacy of total body irradiation and busulfan were studied in recipient preparation for bone marrow transplantation. Male C57BL/6 (B6) mice were prepared for BMT with fractionated TBI or busulfan given in 4 equal doses over 3 days. Both TBI and busulfan are potent stem-cell killers. Both agents resulted in an exponential decrease in CFUs survival with increasing dose down to a 1 x 10(-4) CFUs survival. There appeared to be no break in the curve for either agent. Extrapolated fractional CFUs survival, as related to equivalent donor marrow engraftment or to equivalent 30-day survival without marrow transplantation, appeared lower for TBI as compared to busulfan. This may be due to the effect of busulfan and TBI on different stem-cell populations. Erythroid engraftment was tested after transplanting H-2 compatible LP marrow cells into treated B6 recipients. Greater than 80% of animals demonstrated complete engraftment with 3.4 mg busulfan or 1640 cGy TBI. At these 2 doses, the rate of recovery of donor marrow cellularity and CFUs content in treated recipients were identical for both preparative agents such that a selective effect on the host hematopoietic microenvironment harmful to engraftment was not seen. Complete engraftment in 100% of busulfan-prepared animals could not be achieved as such doses resulted in severe and fatal pulmonary vascular injury at 7-12 weeks posttransplant. PMID- 3043778 TI - Instability of neural xenografts placed in neonatal rat brains. AB - Embryonic mouse retinae placed in neonatal rat brains differentiate normally, form appropriate connections with the host brain, and may survive for longer than 1 year. However, such grafts are susceptible to rejection, either spontaneously or after challenge. Advanced spontaneous rejection of the transplant was identified in about 10% of the animals. In addition, two circumstances have been defined in which mouse retinal grafts can be subsequently induced to undergo rejection. One is following placement of a mouse skin graft on the flank of a rat that has received a retinal transplant in the brain, and the other is following removal of a host eye. After each of these procedures, the neural grafts become infiltrated with lymphocytes and undergo degeneration. It is proposed that this system may provide a useful approach not only for studying the immunology and genetics of neural transplantation, but also for examining the circumstances that precipitate the degenerative events associated with certain autoimmune diseases of the central nervous system. PMID- 3043780 TI - Autoimmunity after allogeneic bone marrow transplantation. A study of 53 long term-surviving patients. AB - Various autoantibodies were screened in 53 long-term survivors after allogeneic bone marrow transplantation. Among them, 40 displayed chronic graft-versus-host disease, with clinical features reminiscent of collagen diseases, especially scleroderma, Sjogren's syndrome, and autoimmune hepatitis. Antinuclear, anti smooth muscle, antimitochondria, anti-liver kidney microsome, and antiepidermal antibodies were found at a frequency of 62.2%, 49.0%, 11.3%, 5.6%, and 11.3%, respectively. The screening for native anti-DNA, anti-extractable nuclear antigen, anticentromere, and anti-salivary gland duct antibodies was negative. The presence or absence of acute GVHD made no difference in the frequency of autoantibodies. No correlation between cutaneous hepatic involvement, sicca syndrome, scleroderma status, and autoantibodies could be established. Despite clinical features mimicking collagen vascular diseases, the biological autoimmune profile of GVHD was different. The precise role of autoimmunity in chronic GVHD remains to be defined. PMID- 3043779 TI - Early function as the principal correlate of graft survival. A multivariate analysis of 200 cadaveric renal transplants treated with a protocol incorporating antilymphocyte globulin and cyclosporine. AB - We examined the factors determining graft survival in 200 consecutive cadaveric renal transplants managed on a quadruple-therapy protocol: Minnesota antilymphoblast globulin, cyclosporine, azathioprine, and low-dose prednisone. Perioperative central venous pressure monitoring and volume expansion were emphasized. To avoid CsA nephrotoxicity in the early posttransplant period, patients were treated with ALG until renal function was established (a mean of 7 days). Therapeutic CsA levels were achieved before ALG was discontinued. Azathioprine was used to supplement CsA in patients with nephrotoxicity or rejection. Twelve-month graft survival was 85% (first transplants 86%, retransplants 79%), with patient survival of 95%. ALG was not associated with excessive clinical cytomegalovirus infections, which occurred in 5% of patients, or with malignancy. When 3 technical failures were excluded, an analysis of numerous factors in the pretransplant and peritransplant period revealed that the strongest correlate of one-year graft survival was early renal function. Grafts with delayed function (DF) had 75% survival, compared with 91% for grafts with good early function (EF). A multivariate analysis confirmed this association: the relative risk of graft loss was increased 2.86 times for DF compared with EF. The mechanism of the deleterious effect of DF was apparently multifactorial: the DF group, by definition, contained all the kidneys that never functioned, but some risk also persisted in kidneys that achieved function. One reason for this may be that DF kidneys that achieved function had higher mean serum creatinine values at 1 month: elevated serum creatinine values at 1 month were strongly associated with increased risk of graft loss regardless of initial function. There was also a higher number of rejection episodes diagnosed in the DF group. These observations suggest that early renal function is a major determinant of graft outcome and should be a target for efforts to further improve renal graft survival. PMID- 3043781 TI - A simple and reliable way of differentiating acute rejection from cyclosporine nephrotoxicity in renal transplantation. PMID- 3043782 TI - Norfloxacin-cyclosporine interaction. PMID- 3043783 TI - Application of a new synthetic absorbable cuff material to vascular anastomosis in liver grafting. PMID- 3043784 TI - Liver transplantation following severe liver trauma. PMID- 3043785 TI - The effect of donor-specific and third-party transfusions on graft survival in haplomatch renal transplants. PMID- 3043786 TI - Short-course total-lymphoid irradiation combined with total-body irradiation to facilitate engraftment of T cell-depleted marrow across a major histocompatibility barrier in mice. PMID- 3043787 TI - Protection from graft-versus-host disease in fully allogeneic chimeras by prior administration of T cell-depleted syngeneic bone marrow. PMID- 3043788 TI - The influence of transfusions from unrelated DLA-matched or mismatched donors upon marrow grafts between DLA-identical canine littermates. PMID- 3043789 TI - Orthotopic liver transplantation managed exclusively with autologous blood. PMID- 3043790 TI - Use of UW solution for kidney transplantation. PMID- 3043791 TI - Syngeneic graft-versus-host disease induced by cyclosporine--a reappraisal. AB - Cyclosporine-induced graft-versus-host disease has been described in lethally irradiated rats and mice following syngeneic bone marrow reconstitution. To further study this apparently CsA-restricted phenomenon, we followed reported protocols by administering CsA orally at 50 or 100 mg/kg/day to irradiated, syngeneically reconstituted C57B1/6 mice. No clinical evidence of GVHD was observed for more than 8 weeks after CsA discontinuation. Ear biopsies and circulating immunoglobulin levels 2-4 weeks after stopping CsA failed to demonstrate histological or serological evidence of GVHD, respectively, compared with mice allogeneically reconstituted with Balb/c marrow. To further follow a previous report, CsA 50 mg/kg/day orally or 10 mg/kg/day intraperitoneally was given to normal C57B1/6 mice prior to using their spleen or bone marrow cells for reconstitution of lethally irradiated syngeneic mice. Clinical monitoring and histological examination of recipients 2-5 weeks after reconstitution again failed to confirm GVHD. Thus our results were uniformly negative in attempting to reproduce syngeneic GVHD in mice. Existing data on rats and humans are reviewed, showing that syngeneic or autologous GVHD is not CsA-restricted and that the syndrome could be equated to the chronic form of GVHD found in rats and patients after allogeneic bone marrow transplantation. PMID- 3043792 TI - Cyclosporine-induced pseudo-graft-versus-host disease in the early post cyclosporine period. AB - Cyclosporine-induced pseudo-graft-versus-host disease (CIPGVHD) in syngeneic or autologous rat marrow chimeras has clinical and histologic features closely resembling classic graft-versus-host disease in the allogeneic chimera. We describe here the pathology and immunopathology of the usual target tissues in CIPGVHD developing in the first week following CsA (early group). The findings are constrasted to the CIPGVHD developing during the second week post-CsA (later group). Six of 9 rats in the early group had acute-type CIPGVHD in the tongue, skin, liver, intestines, and mainstem bronchi. In general, the lymphocytic infiltrates in these tissues were in intimate contact with injured epithelial cells. The intestines had multiple apoptotic lesions. Class II antigen was prominent in the tongue mucosa, but only patchy expression was evident in 2 skin biopsies. All of the lymphocytes infiltrating the mucosa were CD8+(OX-8)/CD4 (W3/25) T cells (OX-19+). Most of the lymphocytes in the lamina propria expressed CD4 as well as CD8 markers, suggesting coexpression. In the later group, 6 of 7 rats had chronic-type CIPGVHD (1 with acute and chronic) while 1 rat had no GVHD (P = .02, Fisher's exact test compared with the early group). These animals had features characteristic of established chronic GVHD in the skin, tongue, liver, intestines, and salivary glands. Fibrosis of the dermis and lamina propria was prominent in the skin and tongue. Submucosal fibrosis was increased in the small intestine. The salivary glands had an interstitial infiltrate and fibrosis with loss of ducts and glands. Class II antigen was prominent in the epidermis of the tongue and skin of all rats. The number of lymphocytes infiltrating the mucosa of the tongue was considerably smaller than seen in the early group. More than 90% of these cells were T cells, as detected by OX-19, and expressed both CD4 and CD8 markers. While most lamina propria lymphocytes expressed the CD4 antigen, there were significantly fewer CD8+ cells, consistent with increased numbers of CD8 /CD4+ helper-phenotype cells. The observations indicate that immediately post CsA, the CIPGVHD is primarily acute, with epithelial infiltrates of CD8+/CD4- T cells and lamina propria infiltrates that include double-labeled cells consistent with immature thymocytes. There is a rapid transition to established chronic-type CIPGVHD by the second week. The residual mucosal infiltrate is now dominated by double-labeled T cells or thymocytes while the lamina propria infiltrate has more mature helper-phenotype T cells. Induced RT1.B/D antigen could be important in the pathogenesis of the peripheral tissue manifestations. PMID- 3043793 TI - The induction of tolerance by cyclosporine. PMID- 3043794 TI - Areas for further experimentation to elucidate the immunosuppressive activity of cyclosporine. PMID- 3043796 TI - Experimental transplantation and cyclosporine. PMID- 3043795 TI - Cyclophilin, a primary molecular target for cyclosporine. Structural and functional implications. AB - An understanding of the mechanism of action of cyclosporine requires the identification and functional characterization of its molecular target or targets in the cell. Our laboratory has presented evidence that cyclophilin (CYP), a low molecular-weight (Mr 17,737) basic protein, is the primary cytosolic receptor for CsA. The high affinity of CYP for CsA (Kd 30 nM) and specificity for immunosuppressive cyclosporine analogs implicate CYP as a pivotal regulator of T cell and B cell activation. CYP exists in at least two isoforms, is abundant (0.05% to 0.4% of total protein) in the cytosol, and is ubiquitous in cells and tissues of eukaryotic organisms. Its amino acid sequence is highly conserved and there is strong evidence that CYP is a member of a new multigene family. These features suggest that one or more CYP isoforms must play a crucial role in lymphocyte activation and perhaps a multifunctional role in cellular physiology. The ability of CsA to suppress expression of several lymphokine and proto oncogene products via a transcriptional control mechanism suggests that CYP may function at some level in a signaling pathway linking membrane receptor stimulation to gene regulatory elements in lymphocytes, and possibly nonlymphoid cell types as well. PMID- 3043797 TI - A comparison of cyclosporine binding by cyclophilin and calmodulin and the identification of a novel 45 KD cyclosporine-binding phosphoprotein in Jurkat cells. AB - Cyclosporine mediates its immunosuppressive effect by preventing the synthesis of lymphokine mRNA during the process of T lymphocyte activation. Although the detailed molecular mechanism by which CsA achieves this effect is unknown, two proteins have been identified as putative intracellular CsA-receptor proteins. One of these, calmodulin, is an important Ca++-binding protein and enzyme cofactor and the other, cyclophilin, is a novel protein that is reported to have protein kinase activity. In this study the CsA-binding capacity of both these proteins has been assessed using CsA-coated ELISA plates and CsA-affinity gel matrices. CsA binding was shown by cyclophilin whereas no CsA-calmodulin binding could be detected under identical conditions. However, it was not possible to demonstrate any cyclophilin-associated protein kinase activity. Jurkat cells were probed for the presence of CsA-binding proteins using the CsA-affinity gel matrix; a 17 KD protein, most probably cyclophilin, was identified as the major CsA-binding protein. In addition, a previously unidentified CsA-binding 45 KD phosphoprotein was precipitated from 32P-labeled Jurkat cells. These results would support cyclophilin as the major, if not only, intracellular receptor protein for CsA. However, the relationship between binding of CsA to cyclophilin and/or the 45 KD phosphoprotein and the immunosuppressive effects of CsA is still unknown. PMID- 3043798 TI - T cell activation in the presence of cyclosporine in three in vivo allograft models. AB - The effect of cyclosporine on a systemic graft-versus-host reaction, cardiac allograft rejection, and a local host-versus-graft reaction in the rat were examined in detail. Therapeutic levels of CsA did not inhibit the early stages of lymphocyte activation but did prevent maturation of the immune response to full effector function--viz., graft rejection or clinical GVH disease. In all three models the phenotype changes in T cells associated with the early stages of activation--i.e., induction of receptors for interleukin 2 (IL-R), induction of MHC class II expression, and coexpression of CD4 and CD8 glycoproteins--were not inhibited by CsA. In the GVH and HVG reactions lymphocyte activation proceeded as far as DNA synthesis. In the systemic GVH model animals showed no sign of GVHD for as long as CsA was administered, but withdrawal of the drug resulted in accelerated lethal GVHD. PMID- 3043799 TI - Regulation of gene expression in lectin-stimulated or lymphokine-stimulated T lymphocytes. Effects of cyclosporine. AB - The effects of cyclosporine were examined on gene expression induced in T lymphocytes by mitogenic lectins and interleukin 2 (IL-2). Used at concentrations that inhibited proliferation of human peripheral blood lymphocytes by approximately 90%, CsA suppressed, to different extents, the phytohemagglutinin stimulated expression of various genes, with levels of mRNAs for IL-2 being inhibited by approximately 100%, c-myc and N-ras by approximately 80%, and c-fos and IL-2 receptors by approximately 50%. Comparisons of the actions of CsA on gene expression in a cloned murine T cell (L2), stimulated with concanavalin A or IL-2, demonstrated that CsA specifically blocked the accumulation of mRNAs for the c-myc and p53 protooncogenes when induced by Con A, but not when induced by IL-2. Taken together, these findings indicate that several pathways can control the expression of a particular gene, and suggest that CsA interferes with only some of these regulatory pathways of gene expression in T lymphocytes. PMID- 3043800 TI - Cyclosporine--relationship of side effects to mode of action. AB - Although cyclosporine has high specificity for the immune system, immunosuppressive therapy with CsA is often complicated by nephrotoxicity. The main morphologic targets of CsA nephrotoxicity include the tubular epithelial and endothelial cells. These cells were investigated in vitro. CsA caused a dose- and time-dependent inhibition of cell growth, vacuolization and fatty change in adherent cells, detachment, and cell death. Inhibition of 3H-TdR incorporation in cells of both tubular epithelial and endothelial origin occurred between 3 microM and 10 microM. Electron microscopy studies revealed cellular swelling, dilatation of the endoplasmic reticulum, and the presence of lipid droplets and giant mitochondria. The content of the main CsA-binding protein, cyclophilin, in these cell-lines was 5-10 micrograms/mg protein and did not differ in various cell lines, including T cells. Immunohistochemistry using rabbit anticyclophilin antibody revealed diffuse distribution of cyclophilin in the cytosol, nuclear membrane, and nucleolus. Whereas lymphoid cell functions are inhibited at 10-100 nM, CsA had no effect on tubular epithelial and endothelial cells at these concentrations. At concentrations of 3-10 microM, CsA caused growth inhibition and cytotoxicity on cells of lymphoid and nonlymphoid origin. Present evidence shows little, if any, relationship of side-effects to the mode of action of CsA. PMID- 3043801 TI - IVth International Workshop on Transplant Aspiration Cytology. October 7-9, 1987, Barcelona (Spain). Proceedings. PMID- 3043802 TI - Consecutive selective aspiration cytology of renal cortex and renal medulla in kidney transplants. PMID- 3043803 TI - Evaluation of needle-core biopsy washings for monitoring rejection in human renal allografts. PMID- 3043804 TI - Identification of virus DNA in kidney transplants by fine-needle aspiration biopsy. PMID- 3043806 TI - Correlation between fine-needle aspiration biopsy and renal biopsy in renal transplantation. PMID- 3043805 TI - Glomeruli in kidney transplants collected by fine-needle aspiration biopsy. PMID- 3043807 TI - Correlation of fine-needle aspirate biopsies with core biopsies after renal transplantation. PMID- 3043808 TI - Comparison of fine-needle aspiration biopsy and tru-cut biopsy performed under ultrasound guidance. PMID- 3043809 TI - The value of needle renal allograft biopsy. PMID- 3043811 TI - Intrarenal inflammation during rejection crises in renal allografts: comparison between fine-needle aspiration cytology and renal biopsy. PMID- 3043810 TI - Relative value of interstitial infiltrates in acute rejection diagnosis after triple-drug immunosuppression: a retrospective morphologic analysis. PMID- 3043812 TI - Diagnostic and prognostic value of HLA-DR expression in fine-needle aspiration cytology in renal grafts immunosuppressed with cyclosporine. PMID- 3043813 TI - Blood eosinophilia and major histocompatibility complex II expression in renal allograft rejection. PMID- 3043814 TI - Major histocompatibility complex class II expression in canine kidney transplantation. PMID- 3043815 TI - HLA-DR antigen expression on tubular and endothelial cells from fine-needle aspiration cytology. PMID- 3043816 TI - HLA-DR expression in fine-needle aspiration biopsy cell samples as a marker of rejection in kidney grafts. PMID- 3043817 TI - Brazilian experience with fine-needle aspiration cytology in kidney transplantation. PMID- 3043819 TI - Aspiration cytology in late cellular rejection. PMID- 3043818 TI - Monitoring of kidney grafts by fine-needle aspiration biopsy. PMID- 3043820 TI - Renal fine-needle aspiration cytology in transplanted children. PMID- 3043821 TI - Statistical and computer evaluation of the cytologic monitoring of kidney graft recipients in the immediate posttransplantation period. PMID- 3043822 TI - A novel use of fine-needle aspiration biopsy: documentation of acute pyelonephritis of the transplanted kidney. PMID- 3043823 TI - Morphologic features in liver transplantation. PMID- 3043824 TI - Aspiration cytology of liver transplants. PMID- 3043825 TI - The total corrected increment in the diagnosis of hepatic dysfunction after orthotopic liver transplantation. PMID- 3043826 TI - Cholestatic syndrome due to preservation injury after liver transplantation. PMID- 3043827 TI - Longitudinal study of major histocompatibility complex antigen expression on hepatocytes in fine-needle aspiration biopsies from human liver grafts. AB - Expression of MHC class II antigens on hepatocytes is part of the natural history of liver grafts. The expression appears to be enhanced by rejection episodes but not during infection and seems to persist on higher levels in chronic rejection and multigraft recipients. Intense expression of class I antigens is a nonspecific feature of the posttransplant situation. PMID- 3043828 TI - Aspiration cytology of liver allografts: monitoring of hepatocyte major histocompatibility complex-DR expression increases accuracy of diagnosis of rejection episodes. PMID- 3043829 TI - Increased hepatocyte cyclosporine A deposits during liver allograft rejection. PMID- 3043830 TI - Transplant aspiration cytology for diagnosis of liver allograft rejection. PMID- 3043831 TI - Liver transplant aspiration cytology is useful for monitoring steroid treatment of rejection. PMID- 3043832 TI - A clear distinction between "immune activation of rejection" and "no immune activation" in liver transplant aspiration cytology. PMID- 3043833 TI - Is the pancreas rejected independently of the kidney after combined pancreatic renal transplantation? PMID- 3043834 TI - Generation of fibroblast and endothelial cell lines from kidney allograft fine needle aspiration biopsy samples. PMID- 3043836 TI - Postischemic injury and acute tubular necrosis: diagnostic and prognostic value of fine-needle aspirates. PMID- 3043835 TI - Cell culture of fine-needle aspirates and tru-cut biopsies. PMID- 3043837 TI - Can incremental scoring of fine-needle aspirates predict histopathologic renal allograft rejection? PMID- 3043838 TI - Analysis of the increment calculation in renal transplant aspiration biopsies. PMID- 3043839 TI - Development of a nomogram to interpret transplant aspiration cytology findings. PMID- 3043840 TI - Practical and technical aspects of fine-needle aspiration cytology. PMID- 3043842 TI - Isolation of islets of Langerhans from the adult pig pancreas. PMID- 3043841 TI - Long-term skin allograft survival by combined therapy with suboptimal dose of cyclosporine and ribavirin. PMID- 3043843 TI - Repair of transplant renal artery stenosis by hypogastric to renal artery fistula. PMID- 3043844 TI - Blood transfusions and HLA compatibility in first cadaveric kidney transplants treated with cyclosporine A. AB - One-year graft survival of 54 first cadaveric kidney transplants that received immunosuppressive treatment with CsA was analyzed with respect to the number of random pretransplant blood transfusions and the HLA class 1 and class 2 matching. Overall graft survival at 1 year was 80.7%. Patients with 3 to 20 pretransplant transfusions had a survival of 93.7% compared with 66.7% in those with less than three or more than 20 transfusions. All kidneys transplanted with two or less HLA A + B mismatches survived at 1 year. With three mismatched antigens survival was of 88.9%. This value was reduced to 66.7% for four incompatibilities. A similar situation was found for HLA-DR matching since all kidneys with full compatibility survived at 1 year compared with 90.9% and 66.7% for one and two mismatches, respectively. HLA-B and HLA-DR exhibited an additive effect since again all grafts with two or less mismatches survived, whereas in the group with three different antigens this figure was 90% and only 1 of the three kidneys with completely different antigens survived. PMID- 3043845 TI - Renal transplantation during pregnancy. PMID- 3043846 TI - International Consensus Development Conference on Anencephalic Donors. January 5, 1987, London, Ontario, Canada. Proceedings. PMID- 3043847 TI - Brain death in the infant and what constitutes life. PMID- 3043848 TI - Donor organs from human anencephalics: a salutory resource for infant heart transplantation. PMID- 3043850 TI - The anencephalic organ donor: affect, analysis, and ethics. PMID- 3043849 TI - Ethical issues in the use of anencephalic infants as a source of organs and tissues for transplantation. PMID- 3043851 TI - Anencephaly--the potential for survival. PMID- 3043852 TI - A self-limiting midline cerebellar syndrome. PMID- 3043853 TI - Urinary bladder stones. Their occurrence in children in South-East Asia. PMID- 3043854 TI - Single layer closure of abdominal laparotomy wounds. PMID- 3043856 TI - A study on chloroquine resistant falciparum malaria in a rural population in Ghana. PMID- 3043855 TI - A model for Plasmodium falciparum sporozoite challenge and very early therapy of parasitaemia for efficacy studies of sporozoite vaccines. AB - Plasmodium falciparum parasitaemias were induced in four non-immune volunteers by the bites of mosquitoes infected from cultured gametocytes. Radical cure was accomplished before three of the four volunteers developed clinical malaria. Despite very low peak levels of parasitaemia, the plasmodium was recultured from the blood of all volunteers. This volunteer model of early detection of parasitaemia and prompt treatment will contribute to the safe and practical efficacy testing of sporozoite vaccines, thus facilitating the selection of candidate vaccines for large scale definitive field trials. PMID- 3043857 TI - Lack of effect of continuous wave ultrasound exposure on in vivo Chinese hamster cheek pouch epithelial mitotic index. AB - Chinese hamster cheek pouches were everted and the epithelia exposed in vivo for 1 min to continuous wave 1.07 MHz ultrasound at intensities ranging from 0.5 to 10.0 W cm-2. Since the mitotic index of the hamster epithelium exhibited a circadian rhythm, the times of sonication and tissue sampling were arranged to allow for detection of a division delay as evidenced by a change in mitotic index in cells insonated in the early S or late S/early G2 phases of the cell cycle. There was no demonstrated effect of the ultrasound exposures on mitotic index. PMID- 3043858 TI - [Injuries of the medial collateral ligament and anterior cruciate ligament of the knee joint and its surgical-functional treatment using the Lemaire technic]. AB - The operative-functional treatment of ligamentous tears of the knee joint avoids additional damage due to immobilisation and shortens the rehabilitation period. The basic idea and details of the Lemaire operations are explained. With an average follow up time of 3.5 years, 113 ligamentous plasties mainly for recent tears of the medial collateral and anterior cruciate ligament have been clinically and radiologically assessed by an independent surgeon. Excellent and good results were obtained in 80 to 90% of the patients. The lateral plasty for torn anterior cruciate ligaments prevents successfully antero-lateral subluxation and thus protects the menisci from tear and the cartilage from degenerative alteration. No obvious increase of torn menisci and degenerative signs could be detected in our series of evaluated knees. The Lemaire procedure for old and recent tears of the medial collateral and the anterior cruciate ligament carries little operative risk and inconvenience for the patient and is indicated in any age group, for the young patient as well as for the older but still active person. We recommend the Lemaire operation as an alternative method in treating ligamentous injuries of the knee joint. PMID- 3043859 TI - [Clinical problems in fracture of the scaphoid bone]. AB - The following specific characteristics of injury cause the problems that occur in scaphoid bone fractures: 1. Mechanisms of accident: There are only a few typical circumstances that cause the injury: as there are false winding of a shrankshaft handle or a fall on the extended hand. In all the other cases the accident is neither realized by the injured person himself or by the physician. As a consequence the possibility of injury of the scaphoid bone is not taken into consideration. 2. SYMPTOMS: In a stable fracture the symptoms do not appear directly but may be clearly delayed or they may disappear very quickly after a short period of intensity, so that the accident is no longer taken any notice of. 3. Radiodiagnostics: Because of the anatomic position of the scaphoid bone any X raying from a lateral view is difficult because the scaphoid is covered by other carpal bones. In case of an undisplaced fracture you often only see very fine fracture lines, which are only to be seen from a direct orthograde view. It is necessary to X-ray the carpus in different positions. 4. THERAPY: If there is the slightest chance of a scaphoid bone fracture an intensive therapy must be directly started. An immobilisation for a too short period may lead to a delayed union and end in a pseudarthrosis. This is also the case if you start the therapy too late. Then the fracture line is already filled up by fibrous tissue. The latest period of time to begin a conservative treatment is six to eight weeks after the day of the accident.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3043860 TI - [Conservative treatment of fractures of the scaphoid bone--indications]. AB - Healing-up of the bone is achieved by immobilization in 96 to 98% of all recent scaphoid fractures. The average immobilization period is eleven weeks. Different methods of immobilization are used. The majority of authors apply an upper arm plaster cast including the thumb for a period of six weeks and then a forearm plaster cast. Even fractures showing a delayed healingup of the bone can be cured in more than two thirds of the cases by prolonged immobilization. In case of dislocated and not reducible fractures as well as fractures with great fragment diastases, conservative treatment with a fist plaster cast should only be performed exceptionally, for example if there is an increased anesthesia risk. In case of negative X-ray findings (four planes) and only clinical suspicion of scaphoid fracture, an initial fracture therapy is recommended with repeated X-ray examination 14 days later. Most of the fractures which have not healed up are vertical oblique fractures within the proximal third, fractures with great diastases, or fractures which have not been immobilized long enough. PMID- 3043862 TI - Dilatation of ureteral orifice for ureterorenoscopy. AB - The history of endoscopic ureteral dilatation began before the turn of the century, and many of those older methods are still in use with the aid of updated instruments. The recent development of controlled hydraulic dilatation promises to make ureteroscopy a one-step procedure. PMID- 3043863 TI - Flexible ureterorenoscopy. AB - The clinical applications of flexible ureterorenoscopes have fallen behind those of their rigid counterparts. Flexible endoscopes that possess a working channel and can also be employed for treatment have at last been developed. This instrument is particularly effective in the treatment of stones in the upper urinary tract, as it can easily follow mobile upper ureteral stones and calculi in the middle or lower renal calices, which are not as accessible with a rigid endoscope. It is important to remember not to exert too much force when inserting the instrument into the ureter, renal pelvis, or renal calices, and, if one remains calm, should the visual field become temporarily obscured, there should be little danger of damaging these structures. This method will probably be increasingly employed, not only for the treatment of stones in the upper urinary tract, but also for the removal of foreign bodies and the diagnosis and treatment of small pathologic conditions or lesions such as carcinoma in situ in which the roentgenologic presentation is unclear. For this expansion of instrument use to be ensured, it is necessary to improve the resolution of the fiberoptic endoscope image and increase the instrument's angulation capabilities, increase the range of accessories that can be employed with it, and also increase familiarity with the instruments on the part of urologists. PMID- 3043861 TI - [Fracture of the scaphoid bone--surgical treatment, indications, method and results]. AB - In fractures of the scaphoid bone, conservative treatment leads to an osseous consolidation in 95% of all cases. A primary surgical treatment seems justified in case of instability by torsion, tilting, or shifting of the fragments, associated fractures of the same arm, as well as de Quervain dislocation fracture. The different surgical possibilities are described and the complications are compiled in a special study. The question if a recent fracture of the scaphoid bone should be managed by conservative or surgical treatment ist not justified considering the rate of 14.5% of complications, i.e. formation of pseudarthroses, observed after surgery of recent fractures of the scaphoid bone. PMID- 3043864 TI - Ultrasonic lithotripsy in the ureter. AB - With the increasing range of indications for extracorporeal shock-wave lithotripsy, "easy" cases of ureteroscopic stone removal have vanished. Clinical experience with ureterolitholapaxy has proved the method safe, effective, and reliable in the hands of the experienced urologist. PMID- 3043865 TI - Second-generation shock-wave lithotripters. Variations, indications, and efficacy in the treatment of ureteral calculi. AB - Second-generation lithotripters offer significant advances in the technology and delivery of high-energy shock waves. The final position that ultrasonographically guided machines will earn in the spectrum of lithotripters can be determined only as more clinical series are reported. Nonetheless, their compromised ability to handle ureteral calculi casts a shadow on the acquisition of such instruments by facilities that cannot afford to own and operate both x-ray and sonographically guided lithotripters. PMID- 3043866 TI - Percutaneous antegrade approach to ureteral calculi. AB - Using the percutaneous techniques described here, antegrade ureteral calculus extraction has been successful in more than 90 per cent of reported cases. Thus, stones in the proximal two thirds of the ureter that cannot be managed with standard retrograde techniques alone or in combination with ESWL or that are associated with large renal calculi can be removed with low morbidity by percutaneous methods and endourologic techniques. PMID- 3043867 TI - Unusual applications of ureteroscopy. AB - Six cases of combined percutaneous and ureteroscopic opening or reanastomosis of completely occluded ureters are described. All patients had previously had nephrostomy tubes inserted to salvage their kidneys. PMID- 3043868 TI - Ureteral stents. Indications, variations, and complications. AB - Indwelling ureteral stents offer the urologist an enormous arsenal against a host of urologic diseases. No stent is ideal, and as such it is incumbent on the surgeon to be familiar with the various indications for usage, selection, modes of insertion, and potential for complications. With such information, the surgeon will optimize the efficacy and safety of this device in the care of patients. PMID- 3043869 TI - Bioeffects of extracorporeal shock-wave lithotripsy. Strategy for research and treatment. AB - Available information on the bioeffects of ESWL is insufficient to characterize the tissue injury induced by shock waves. The cellular mechanisms have not been elucidated, nor are there enough data to establish objective criteria for treatment. PMID- 3043871 TI - Acute and chronic urinary infection: present concepts and controversies. AB - The current approaches to diagnosing the various renal inflammatory diseases are briefly reviewed. Terminology is varyingly used by different individuals. Our current understanding of these disease processes is discussed in an effort to review the current status of both the mundane and the esoteric infections of the kidney. PMID- 3043870 TI - Management of ureteroscopic ureteral injuries. AB - Ureteroscopy will continue to have a place in the management of renal stone disease and is invaluable to evaluate other types of ureteral lesions. Although ureteroscopy can be performed safely in most cases, it is invasive and there is a potential for ureteral injury. The well-trained urologic surgeon should be familiar with techniques to minimize the incidence of complications of ureteroscopy. However, faced with a ureteral complication, no matter the severity, the surgeon should have the ingenuity and expertise to restore anatomic integrity to the renal unit. PMID- 3043872 TI - New and old contrast agents: pharmacology, tissue opacification, and excretory urography. AB - In equivalent doses for intravenous urograms conventional ionic contrast agents give iodine concentrations in the urine of approximately 30 mg iodine/ml, nonionic contrast media provide approximately 50 mg iodine/ml, and the ionic dimer Hexabrix approximately 70 mg iodine/mL. These new low osmolality, contrast media provide significantly higher urinary iodine concentrations than conventional ionic contrast media, provide better diagnostic quality excretory urograms, better patient tolerance, and fewer adverse side-effects and serious reactions. These new low osmolality, contrast media have significant advantages in intravenous urography in both safety and efficacy when compared to conventional higher osmolality contrast media. PMID- 3043873 TI - Renal tuberculosis. AB - Imaging of renal tuberculosis is reviewed, with the observation that early changes are best seen with urography/pyelography and chronic changes are best evaluated with ultrasound/computed tomography. Imaging procedures are of limited value until a proven bacteriologic diagnosis is made. PMID- 3043874 TI - Renewed role of nuclear medicine in renovascular hypertension. AB - The role of nuclear medicine in the differential diagnosis of renovascular hypertension has been subject to considerable disagreement. The renogram has several distinct advantages as a diagnostic test, among which are the following: it is safe, easily performed, and has a high sensitivity for the differentiation of normal renal function from abnormal renal function. The use of the captopril renogram is based on the observation that renal function appears to be maintained in renovascular hypertension through a component of the renin-angiotensin system. Captopril renography will greatly improve the efficacy of the radionuclide renogram in the differential diagnosis of renovascular hypertension. PMID- 3043876 TI - New and old contrast agents: physiology and nephrotoxicity. AB - Newer contrast agents have recently been developed to improve patient safety and imaging quality. This survey examines renal handling of contrast media, mechanisms of contrast enhancement and questions related to renal toxicity. Methods to minimize contrast-medium-induced acute renal failure are suggested. The introduction of low osmolality contrast media promises reduced patient risk and improved renal enhancement. PMID- 3043875 TI - Urinary extracorporeal shock wave lithotripsy: equipment, techniques, and overview. AB - Second generation urinary lithotriptors are characterized by extensive technical alterations and significant equipment improvement in the functional, logistical, and medical aspects of shock wave lithotripsy (SWL). These newer devices feature a water bath-free environment, a reduced anesthesia requirement, improved imaging, functional uses in addition to lithotripsy, or combinations thereof. Shock wave generation by spark gap, electromagnetic, piezoelectric and microexplosive techniques are related to their peak energy, frequency, and total energy capabilities which impacts on both anesthesia needs and the length and number of treatment sessions required to pulverize calculi. A master table summarizes the types of SW energy, coupling, imaging systems, patient transport, functional features, cost, and treatment effectiveness of 12 worldwide lithotriptors in various stages of investigative and clinical trials as monitored by the Food and Drug Administration (FDA) of America. PMID- 3043877 TI - [Chemoprevention in superficial bladder cancer 1988]. PMID- 3043878 TI - [Urinary diversion following radical cystectomy: ileal conduit or ileal replacement bladder?]. AB - Among those patients who have undergone radical cystoprostatectomy in the Department of Urology at the Municipal Hospital of Kassel (FRG) since 1985, 57 received a continent ileal bladder anastomosed to the membranous urethra. In a retrospective study intra- and postoperative complications, late complications and prognosis of the tumor disease of these patients were compared with those who had an ileum conduit from 1983 to 1988 (n = 44). The intra- and postoperative complications were nearly identical. During the further postoperative course, patients with a continent ileal bladder may present with strictures at the anastomosis of the ileal bladder and urethra, incomplete voiding with residual urine, secretion of water by the mucosa, and metabolic disturbances because of absorption of substances usually eliminated with the urine. The latter decreases with time as the glandular epithelium of the ileal mucosa changes into mucigenous cells with villous atrophy. The danger of local tumor recurrence and metastasis is the same in patients with an ileal bladder and an ileal conduit. PMID- 3043879 TI - [Selective microsurgical therapy in vascular-induced erectile impotence]. AB - Patients with erectile dysfunction should be subjected to a complex and complete diagnostic procedure, including selective pharmaco-phalloangiography and dynamic pharmacocavernosonography only when SKAT has failed and after the exclusion of neurological disorders. Patients being offered these invasive procedures should be highly motivated and willing to undergo surgical correction of the vascular origin of their erectile impotence. If peripheral vascular occlusive disease is found affecting the pelvic arteries, the best operative technique must be selected. In our opinion, this decision should be made intraoperatively after visualization of the dissected dorsal penile vessels. Use of the Doppler probe can be extremely helpful during the operation. Intraoperative findings on the degree of arteriosclerosis and arterial flow can differ from those allowed by preoperative diagnostic procedures. Penile erectile function was restored in seven of nine patients after penile revascularization performed under microsurgical conditions. We used the operative methods described by Virag and Hauri, applying our own modification of the Hauri procedure in two patients. With careful selection of patients and methods it should be possible to resolve individual patients' problems with peripheral occlusion, even though the exact hemodynamic pathology remains obscure. PMID- 3043880 TI - Pathogenesis of recurrent urinary tract infection: use of understanding as therapy. AB - Understanding bacterial adherence and explaining it to patients is essential in the management of recurrent urinary tract infection. Bowel flora, usually Escherichia coli, colonize first the vaginal introitus, then the urethral mucosa, and they subsequently ascend to the bladder. The essential step is bacterial adhesion to receptor sites on uroepithelial cells. Women who suffer recurrent infections have more receptive cells than the cells of other women. Long-term antimicrobial therapy reduces susceptibility to reinfection. Bringing patients into an understanding of their infectious process and into a partnership in the alleviation of their suffering is a powerful therapeutic tool. PMID- 3043881 TI - Protocol for diagnosis of urinary tract infection: reconsidering the criterion for significant bacteriuria. AB - The traditional criterion of 10(5) colony-forming units (CFU) per milliliter of urine to diagnose urinary tract infection was based on studies of pregnant and nonpregnant women with asymptomatic bacteriuria or acute pyelonephritis. Recent studies of symptomatic women revealed that urine cultures in approximately one third of those with confirmed urinary tract infections grew only 10(2) to 10(4) CFU/mL. The major causes of acute dysuria among such women are urinary tract infection, sexually transmitted disease, and vaginitis. In most instances, it is possible to make the diagnosis based on clinical features. The major features of urinary tract infection are internal dysuria; frequency, urgency, and voiding of small volumes; abrupt onset; suprapubic pain; presence of pyuria. Presence of hematuria which occurs in about 50 percent of patients strongly suggests bacterial cystitis. Three to seven days of empiric antimicrobial therapy is indicated for these patients, with selection of a first-line antimicrobial agent that offers efficacy against Escherichia coli or Staphylococcus saprophyticus; reasonable cost; few side effects. Ampicillin is not recommended. Indications for culture include uncertain clinical features; history of previous infection within the past three weeks; duration of symptoms of more than seven days; recent hospitalization or catheterization; pregnancy; diabetes. To maximize the sensitivity and specificity of the urine culture in acutely symptomatic women, it is necessary to request the laboratory to report 10(2) to 10(4) CFU/mL. PMID- 3043883 TI - [Evaluation of the condition of an allotransplanted kidney during the treatment with Sandimmune]. AB - Instrumental-functional examinations (echography, rheography, electrothermometry) were performed in 22 patients with the allotransplanted kidney treated by Sandimmune and in 20 patients treated by standard immunodepression (Azathioprine and corticosteroids). An analysis of the examinations performed has shown reliable differences between the complications which involved the transplant size, cortical layer, skin-temperature gradient, the degree of the venous wave. PMID- 3043882 TI - [Stable metal osteosynthesis in ununited fractures and pseudarthroses of the tibia]. AB - The plunge nailing osteosynthesis with the Kuntscher nail through the tibia tuberosity and with the Bogdanov nail through the medial malleolus provide stable fixation of the fragments. When combined with decortication of the bone diaphysis and autoplasty with translocated bone transplants the method gives consolidation in ununited fractures and false joints of the tibia within 17-21 weeks. The method was used in operations of 20 patients. The follow-up observation of 18 patients lasted longer than a year. All of them resumed their working capacity. PMID- 3043884 TI - [Treatment of closed injuries of the urethra in pelvic trauma]. PMID- 3043885 TI - [Endoscopy of the small intestine]. PMID- 3043887 TI - [Peripheral hemodynamics in patients with arteriosclerosis obliterans and arteritis studied by ultrasonic dopplerography]. PMID- 3043886 TI - [Ultrasonic examination in cholecystitis and biliourolithiasis]. AB - Results of the ultrasonic examination (USE) in 511 patients with cholecystitis and biliurolithiasis are described. The authors point to possible use of USE on urgent indications for differential diagnosis of acute cholecystitis. The diagnostic signs of biliary and urinary stones are divided into the absolute and relative ones. PMID- 3043889 TI - Treatment of calf E coli enterotoxaemia. PMID- 3043888 TI - Tuberculosis in imported red deer (Cervus elaphus). AB - An outbreak of tuberculosis due to Mycobacterium bovis in farmed red deer imported from an eastern European country is described. Twenty-six of the 106 deer examined at autopsy were found to be infected and 19 had visible lesions of tuberculosis. Single comparative intradermal tuberculin tests on 51 deer showed that the test had a specificity of 61.3 per cent and a sensitivity of 80 per cent relative to subsequent biological and cultural tests on tissues taken at autopsy. Three hundred and seventy eight culled fallow and sika deer which had been running in a park in contact with some of the infected animals were found to be free of tuberculosis. PMID- 3043890 TI - Embryo transfer: its uses and recent developments. AB - The technique of embryo transfer is now routinely employed in several species and with several objectives. In laboratory animals the technique is used mainly in investigations of reproductive biology, whereas in human beings it is used to overcome specific forms of impaired fertility. Because embryo transfer combined with superovulation of the donor can significantly increase the female reproductive rate its greatest application to date has been in farm animals, in which it is widely used in both research and commercial production. Within the past few years there have been many advances in the techniques used in farm animals, particularly in the area of embryo manipulation. The supply of embryos can now be increased by repeated superovulation and by embryo bisection and there has been significant progress in in vitro fertilisation technology. Deep freeze ( 196 degrees C) storage of embryos is now routine and may be combined with direct one-step thawing and removal of cryoprotectant. This technique allows the routine non-surgical transfer of embryos in the field. There is also evidence that a routine non-traumatic procedure for sexing embryos may soon be developed. Identical multiplets or clones have been produced by the microdissection of embryos and more recently by the transfer of nuclei from embryos into unfertilized oocytes. Transgenic animals have been produced by the microinjection of recombinant DNA into one of the pronuclei of single-cell unfertilized ova. PMID- 3043891 TI - The role of bandaging in the management of open wounds. PMID- 3043892 TI - [A case of ultrasonic diagnosis of pseudomyxoma of the peritoneum]. PMID- 3043893 TI - [Use of echography in ambulatory examination conducted in a general hospital]. PMID- 3043895 TI - Bilateral synchronous germ cell carcinomas. Case report, review of literature. PMID- 3043894 TI - Chemical crosslinking of bacteriophage phi 6 nucleocapsid proteins. AB - phi 6 is a lipid-containing dsRNA bacteriophage of Pseudomonas syringae. Its nucleocapsid (NC) has common features with Reoviridae core particles. We report here the crosslinking of phi 6 NC proteins with cleavable 12-A span chemical crosslinker, dithiobis(succinimidyl propionate). The crosslinked complexes were analyzed in two-dimensional polyacrylamide gels or by using monoclonal antibodies to uncleaved protein complexes in one-dimensional protein gels. The NC surface protein (P8) forms a series of multimeric homopolymers. The phi 6 lytic enzyme, protein P5, is associated with P8 on the NC surface. The interior NC proteins P1 and P4, associated with the virus polymerase activity, are also in contact with the P8 shell. A P1 + P4 complex is also formed. Only one of the NC proteins (P7) did not easily form complexes with the other NC proteins. These results indicate a very closely packed P8 surface lattice with specific contacts to the internal NC proteins. PMID- 3043896 TI - Prostacyclin production following in vitro mixing of normal with hemolytic-uremic syndrome serum. AB - Serum from patients with the hemolytic-uremic syndrome (HUS) usually has a diminished ability to support the production of prostacyclin (prostaglandin [PG] I(2)). An impaired ability to produce this potent antiaggregatory substance could account for the thrombotic microangiopathy that is characteristic of the syndrome. We did in vitro mixing experiments to determine if adding normal serum in various concentrations would improve the ability of HUS serum to support PGI(2) production when incubated with cultured human endothelial cells. Mixing normal with HUS serum in a 1:2, 1:3, and 1:6 ratio generally enhanced the PGI(2) supporting capacity of the HUS serum. Moreover, adding normal serum yielded a mixture whose supporting capacity was between the normal and the HUS serum's value, and the PGI(2)-supporting capacity could be predicted by calculating the weighted average value of the components of the mixture. There was a strong correlation between the calculated (predicted) and the actual experimental values (r = .95, P<.001). PMID- 3043898 TI - Complications of gastroesophageal reflux. AB - An edited summary of an Interdepartmental Conference arranged by the Department of Medicine of the UCLA School of Medicine, Los Angeles. The Director of Conferences is William M. Pardridge, MD, Professor of Medicine. Several specialists have recently recognized that gastrointestinal reflux causes complications resulting in significant disease. It causes discomfort, indigestion, esophagitis, Barrett's esophagus, and carcinoma of the esophagus. Pediatricians attribute many early pulmonary problems, and even some sudden deaths in infants, to the reflux of gastric contents. Otolaryngologists now recognize that many cases of nonbacterial, nonspecific pharyngitis and laryngitis are due to the reflux of gastrc acid secretions. Contact granuloma and cancer of the larynx may, in some instances, be secondary to nocturnal reflux. Thoracic surgeons and pulmonologists believe chronic tracheobronchitis and some cases of pulmonary disease are attributable to recurrent bathing of the respiratory epithelium by aspirated gastric contents. An awareness of the many complications of gastrointestinal reflux should lead to a multidisciplined attack on the factors responsible for these diseases. PMID- 3043899 TI - Stratifying risk after a myocardial infarction. AB - These discussions are selected from the weekly staff conferences in the Department of Medicine, University of California, San Francisco. Taken from transcriptions, they are prepared by Drs Homer A. Boushey, Professor of Medicine, and David G. Warnock, Associate Professor of Medicine, under the direction of Dr Lloyd H. Smith, Jr, Professor of Medicine and Associate Dean in the School of Medicine. Requests for reprints should be sent to the Department of Medicine, University of California, San Francisco, School of Medicine, San Francisco, CA 94143. PMID- 3043897 TI - Modern neurosyphilis--a critical analysis. AB - Neurosyphilis remains a source of perplexity for today's physicians. Controversies exist over the interpretation of serologic tests, cerebrospinal fluid (CSF) abnormalities, diagnostic criteria, and treatment regimens. Its occurrence with human immunodeficiency virus (HIV) infection has raised fears of its recrudescence. A critical analysis of the evidence behind these viewpoints leads to several conclusions: the CSF VDRL is the most appropriate diagnostic test; pleocytosis is the only reliable CSF measure of disease activity; commonly accepted diagnostic criteria do not exclude nonsyphilitic disease; and treatment requires the prolonged use of parenteral penicillin, but no superior regimen has been found. Most data do not currently support the view that concurrent HIV infection produces accelerated or resistant neurosyphilis. PMID- 3043900 TI - [Complications of the treatment of hydrocephalus in children by ventriculoperitoneal shunt]. PMID- 3043901 TI - [The usefulness of bacteriological examination of pharyngeal smears]. PMID- 3043902 TI - [Epithelial neoplasms of the external auditory meatus]. PMID- 3043903 TI - [Anti-inflammatory factors in human milk]. PMID- 3043904 TI - [Antibiotics and drugs active against abdominal infections in newborn infants]. PMID- 3043905 TI - [Current status of studies on the epidemiology of epilepsy]. PMID- 3043906 TI - [Epidemiological basis of the prevention of disease]. PMID- 3043907 TI - [General practitioner Marian Jozef Skowronski, author of the first manual on herbal medicine for children (1900-1979)]. PMID- 3043908 TI - [New endoscopic methods in the diagnosis and treatment of diseases of the digestive tract]. PMID- 3043909 TI - [Magnetic resonance tomography as a localization method in hyperparathyroidism]. AB - Magnetic Resonance Imaging (MRT) was performed in 36 consecutive patients with hyperparathyroidism. MR tomograms of 31 patients were evaluated and compared with the results of operation and histology (n = 29). In the remaining 5 patients MR examination was not completed, due to claustrophobia or motion artefacts. MR examinations were performed in 2 superconductive magnets (0.5 and 1.5 Tesla). A surface coil with following spinecho sequences was used: SE: TR/TE: 550-700/15 30; 2000/22-100. All patients were subjected to additional sonography. Out of 28 parathyroid adenomas 25 were identified on MR tomograms (sensitivity: 73%, specificity: 90%). However, only 2 out of 6 hyperplastic parathyroid glands were localized on MR tomograms. Lesions missed on MR tomograms measured 15 mm and less in diameter. It is characteristic that parathyroid adenomas showed isointense MR signal to the thyroid (SE 550/30 and hyperintense MR signal to fat (SE 2000/100). Different signal intensities of the adenomas were observed in 25% of the cases. MR imaging is a valuable diagnostic method for preoperative localisation of parathyroid adenomas. We think that MR imaging should be performed when sonography and subtraction scintigraphy are not able to identify a suspected adenoma in the same location. PMID- 3043910 TI - [3-dimensional scintigraphy of the brain: single photon emission computerized tomography]. AB - Both IMP and HMPAO are lipophilic substances with high blood brain barrier permeability and sufficiently long constant retention in the brain parenchyma to enable SPECT to be carried out with a computer controlled Gamma camera. Both tracers are almost completely cleared from the blood during a single passage through the brain. Hence, the regional distribution of activity in the brain is proportional to the regional cerebral blood flow. Computerized acquisition and data processing result in 16 mm thick tomograms at a spatial resolution of 12 mm, which enable clinicians to detect and characterize the pathogenesis and the development of cerebral disorders in terms of altered function. At present, SPECT can be used in daily clinical routine (final SPECT images are available 30 min after the investigation) in cerebrovascular diseases and, because of the coupling of metabolism to cerebral blood flow, in disturbances of cerebral blood flow due to pathological changes of brain metabolism, i.e. in epilepsy and in degenerative disorders. The forthcoming application of physiologically highly specific tracers (e.g. receptor ligands) as a means of studying the distribution of receptors and drugs in the brain will further enlarge the eventual role of SPECT in clinical management. PMID- 3043911 TI - [Is type I diabetes a virus-induced disease?]. AB - The conditions are critically reviewed under which diseases might be aetiologically related to infection by a certain virus. Such a causal correlation has to obey Koch's postulates, but may be very difficult to prove in the case of a "hit and run" process. This will be exemplified in the case of insulin dependent diabetes mellitus (IDDM). Observations in humans and the results of experiments on laboratory animals are reported, whereby the Coxsackie B and mumps viruses are of particular interest. Furthermore, the mechanisms by which viruses may produce autoimmune diseases are discussed, including virological and immunological aspects. The hypothesis of "molecular mimicry" by Oldstone is quoted as a unifying one, allowing the combination of both aspects. His main assumption is oligopeptide homology between certain virus proteins and some cell proteins and some examples are given. In the presence of such homologies the immune system is first stimulated by the parasitic virus protein, but somewhat later this reaction switches against the host structures, causing continuing cellular damage with the development of autoimmune disease. It is concluded that Koch's postulates in such cases have to be supplemented by assaying for amino acid homologies in viruses and certain cell types. PMID- 3043912 TI - [Conventional or functional insulin therapy?]. AB - The main difference between "conventional" and "functional" insulin replacement is that the former requires meals to be taken at set times throughout the day to avoid hypoglycaemic insulin reactions, while the latter separates insulin replacement in the basal ("fasting") state from that required with food intake. Such strategy reverses conventional insulin treatment (namely balancing the action of administered insulin by a fixed dietary intake), by substitution with a functional control of hyperglycaemia on the basis of tailored insulin doses. To this end blood glucose self control and systemic blood glucose correction are a must during functional insulin substitution, but not necessarily so during conventional insulin therapy. From this it is apparent that "conventional" and "functional" insulin therapy refer to different strategies, both of which may be intensified by more strict rules, although the term "intensified" remains without any conceptional meaning per se. However, whatever the therapeutic recommendation, the attending physician has to be aware that he must appropriately inform and train the insulin-deficient patient (a) on how to deal with a proposed treatment schedule, and (b) to the point that he fully understands the available therapeutic possibilities and the difference in their quality. Experience has shown that a majority of informed patients opt for functional therapy. PMID- 3043913 TI - [Principles of immunotherapy in insulin-dependent diabetes mellitus]. AB - There is now convincing evidence that insulin-dependent (Type 1) diabetes mellitus is an immunological disease. This is derived from observations of a genetically determined predisposition, an association of recent-onset diabetes with viral infections, an inflammatory cell infiltrate affecting the islets of Langerhans, autoantibodies against islet cells, insulin, insulin receptors, and other organ-specific or non-organ-specific antigens, as well as abnormalities of cell-mediated immunity and of non-antigen specific mediators. Finally, recurrent diabetes in cases of pancreas transplantation in monozygotic twins discordant for insulin-dependent diabetes underlines the influence of an autoimmune insulitis. The current concept of the aetiopathogenesis of most cases of insulin-dependent diabetes is that in genetically susceptible individuals any form of damage to beta cells by viral, toxic, dietary or other environmental factors may initiate beta cell destruction and/or aberrant antigen expression, followed by a self perpetuating, mostly cell-mediated autoimmune destruction of the insulin producing cells. Successful immunoprevention in autoimmune diabetes models, on the basis of these recent concepts, led to the assumption that immunotherapy by means of immunomodulative or immunosuppressive drugs might be a possible tool in the treatment of patients with recent onset insulin-dependent (Type 1) diabetes mellitus. PMID- 3043914 TI - [The status of managing the type I diabetic patient in Austria. Organization]. AB - During the Annual Meeting of the Austrian Diabetes Association various diabetic centres presented their success and failure rates with respect to metabolic control and prevention of late complications in diabetic patients on a country wide basis. The analysis revealed that only 30% of all type I diabetic patients are adequately controlled. Intensive education in diabetes self-management, capable of leading to optimal metabolic control in up to 50% of the instructed patients, is however available only to a small minority of the diabetic patients to date. Despite the enormous improvement which has been achieved in the management of pregnant diabetic women, intensive specialized care is often commenced far too late. In rural areas, in particular, even conventional therapy is not fully implemented and late complications are, thus, inevitable. PMID- 3043915 TI - [Acute and long-term psychological problems following hypoglycemias]. AB - 1. Detection of hypoglycaemic episodes is of psychiatric importance with respect to acute therapy, long-term treatment, which includes consideration of changes in specific behaviour patterns, and also forensic questions. 2. All possible therapeutic efforts must be focussed on the avoidance of repeated hypoglycaemic episodes to ensure the prevention of organic psychosyndromes (dementia). 3. The therapeutic proceedings should consider methods of behavioural medicine and behavioural modification too. Such techniques may amplify the traditional medical aspects, especially stressing the significance of compliance, illness-related information, family variables, diet, and bodily exercise (according to Epstein). Methods of behavioural medicine have shown that specific techniques should be applied with care and take individual variability and conditions into account according to single subject methodology. Uncritical application of such techniques is useless. PMID- 3043916 TI - [Results of cyclosporin A therapy in the early phase of type I diabetes mellitus]. AB - A number of findings concerning the pathogenesis of insulin-dependent diabetes mellitus have shown that an autoimmune process is responsible for the destruction of the beta-cell mass, and that a major part of this process has already occurred during the prediabetic phase of the disease. Various immunosuppressive intervention trials have, thus, recently been performed. Remission rates of between 30% and 50% in the Canadian cyclosporin A (CyA) pilot study prompted two placebo-controlled double-blind studies applying this medication. In the French CyA trial 122 patients were followed up for 9 months. 37% of those on high-dose CyA (whole blood levels greater than 300 ng/ml) achieved total remission, compared with 16.7% of those on low-dose CyA (blood level less than 300 ng/ml) and 5% of the placebo group. The Canadian-European trial included 188 patients, of whom 42 were treated in the Viennese centre. Diabetes had been diagnosed in these 42 patients not more than 6 weeks previously, and the duration of their symptoms did not exceed 14 weeks. Whole blood CyA levels ranged from 400 to 800 ng/ml. In relation to short duration of symptoms and early commencement of treatment up to 10 times higher total remission rates were found in the CyA group as compared with the placebo group. In both studies similar side effects were seen. Apart from the cosmetic side effects (hypertrichosis, gingival hyperplasia) a decrease of 20% in creatinine clearance and an increase of 20% in plasma creatinine level seemed to be of clinical importance.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3043917 TI - [Disorders of erectile potency in diabetes mellitus]. AB - The incidence of erectile dysfunction in patients with long-term diabetes mellitus can be as high as 50%. Diabetic microangiopathy is regarded as the most important pathogenic factor. In this review of a group of 210 impotent patients evaluated and treated at our centre we report on the examination data (angiopathy, neuropathy, psychogenic factors) in 36 patients with diabetes in comparison with the corresponding findings in 169 non-diabetic patients. In 5 patients erectile dysfunction had actually preceded the clinical manifestations of diabetes mellitus. Autoinjection therapy was started in 62% of all the diabetic patients, since this is effective, minimally invasive and, therefore, applicable to a large group of patients. This form of therapy was accepted by 90% of our patients' partners, which is in accordance to the reports in the literature. However, treatment had to be interrupted in 2 out of the 22 patients, owing to lack of cooperation on the part of the sexual partner. No complications were attributable to autoinjection therapy. PMID- 3043918 TI - [Serum bile acids in diffuse liver parenchymal damage and their relation to ultrasound diagnosis]. AB - 40 patients with sonographically detected diffuse liver diseases of different severity and 8 controls with normal ultrasound of the liver were evaluated for fasting concentrations of serum bile acids (SBA) using an enzymatic method. Differences of SBA levels were significant among 4 groups classified as to severity by ultrasonography; SBA levels above 4.6 mumol/l were specific for the more severe grades of liver disease. Measurement of SBA and the evaluation of echographic patterns may be helpful in the decision to perform a liver biopsy. PMID- 3043919 TI - [Tumor markers. Their current diagnostic status]. AB - Many tumor markers have been described in the hope of finding a blood test for cancer, and some have found their way into widespread but indiscriminate clinical use. Classically, a marker is synthesized by the tumor and released into the circulation, but it may be produced by normal tissue in response to invasion by cancer cells. Several characteristics define the ideal tumor marker: It should be produced by the tumor cell and be readily detectable in body fluids, not present in health or benign disease, reflect the bulk of the disease and correlate with the results of anticancer therapy. No marker described to date meets all of these criteria. Application of monoclonal antibodies to detect tumor markers promises to greatly expand the number of diagnostic tests available. Monoclonal antibodies have found to provide a degree of specificity not possible using conventional heterogeneous population of antibodies. Antibodies to tissue specific markers are being used to label tissue sections by immune labelling techniques such as immunofluorescence and immunoperoxidase. Identification of a particular marker in the tumor cells of a metastatic lesion may aid in determining the primary site of a tumor. Furthermore radiolabelled antibodies have been used to localize antigen bearing tumors. Some occult neoplasms have been localized by this procedure, however the practical usefulness remains to be determined. There is still a need for new markers, specific for certain tumoral tissues. Perhaps by aid of monoclonal antibodies it will be possible to identify molecules having a functional significance for tumor-growth (e.g. oncogene products). Such markers may circulate in ultra small amounts, but may still have significance for the monitoring of patients. The continuing improvements of tumor-associated markers may help to approach in cancer treatment and diagnosis. PMID- 3043920 TI - [Therapy of advanced prostatic cancer]. AB - Prostate cancer is a polyclonal tumor. There are about 70 bis 75% responders after hormone therapy among previously untreated patients in EORTC trials. Therefore 25 to 30% of the tumors are resistant to androgen. Surgical and hormonal castration show similar results. After progression under hormone therapy response can be reached with antiandrogenic therapy in 30 to 38%. All this leads to the conclusion that it is necessary to base treatment on tumor adaptation and selection. Cell clones insensitive to androgen therefore need cytostatic chemotherapy. It is not yet proofed whether polychemotherapy really shows better results than monotherapy does. Following the thesis of a polyclonal tumor, therapy of advanced prostate cancer must be based on hormone therapy and cytostatic chemotherapy. Prostate specific antigen is a marker for an early recognition of progression. PMID- 3043922 TI - [Tocolysis in the first half of pregnancy]. AB - This study includes a total of 60 gravidae with a pregnancy between 8 and 17 weeks of gestation and the signs of threatened abortion (contractions and bleeding). The sonographic examination had shown in 14 patients no gestational sac in the uterus; in 6 patients a deformed gestational sac; in 5 patients a fetus without signs of vitality. In all these patients curettage was performed. In 32 gravidae a therapy seems to be successful: in terms of bedrest and the administration of 0.33 mcg/min Hexoprenaline as a continuous infusion. In only 3 patients an abortion occurred before the end of the 20th week of gestation. In 29 gravidae pregnancy continued undisturbed. PMID- 3043921 TI - [Serology in HIV infection: comparison of indirect immunofluorescence, Western blot and enzyme immunoassay]. AB - 145.990 sera obtained from AIDS-risks groups, hospitalized patients, blood donors etc. were tested for the presence of antibodies against the AIDS-Virus (HIV:Human Immunodeficiency Virus). All sera were submitted to ELISA screening. Sera with positive and questionable results were submitted to two independent confirmatory tests (Western Blot and immunofluorescence). In case of discordant confirmatory tests sera were additionally tested with the Abbott anti-HIV envelope/anti HIV core ELISA. The results of the present study demonstrate: 1. HIV serology has been definitely improved during the last year due to the development of high quality reagents. 2. Provided skilled and trained personnel the combined use of HIV confirmatory tests (Western Blot and immunofluorescence) does not give false negative or false positive results. 3. The results of both confirmatory assays may not always be interpretable ("problem sera"); this occurs more often with the Western Blot technique than with immunofluorescence. 4. Discrepant results in confirmatory tests necessitate the evaluation of many additional tests as possible. Such persons or patient should be further controlled serologically and clinically. 5. Checking the "problem sera" for antibodies against HTLV I gave only one single positive result in a Japanese. PMID- 3043923 TI - [Primary diagnosis in acute adnexitis]. AB - Though acute salpingitis is the most frequent gynecological illness of a young woman, it occurs nevertheless less frequently than it is diagnosed. The reason for this is to be found above all in the fact, that the criteria of the findings are judged too superficially. If simple anamnestic, clinical or biochemical parameters such as the characteristics and duration of the pain, the findings of gynecological examination, BSR and considerations about differential diagnosis were rated more precisely, considerably fewer cases would have to be assigned to laparoscopy and the elucidation of the indistinct abdominal pains could be started more efficiently. PMID- 3043927 TI - [Possibilities and trends in pharmacotherapy of cardiac arrhythmias in childhood and adolescence. 2: Anti-arrhythmia agents and anti-arrhythmia active drugs in the present and future]. PMID- 3043924 TI - Cocaine: analysis, pharmacokinetics, and metabolic disposition. AB - The ability to measure concentrations of cocaine in body fluids can contribute substantially to any investigation of cocaine's pharmacological effects. Design of research which involves the administration of cocaine must take into account current knowledge regarding the drug's pharmacokinetics. Cocaine's very rapid elimination from the body should be considered in attempting to understand patterns of cocaine abuse, and such phenomena as bingeing and acute tolerance. Accurate analysis of cocaine and/or its metabolites is essential to the diagnosis and evaluation of cocaine use whether for medical or forensic purposes. Appropriate selection of methods for analysis of cocaine depends upon the intended purpose of the assay, and correct interpretation of the data obtained upon knowledge of cocaine's kinetics and metabolic disposition. PMID- 3043925 TI - Evolving conceptualizations of cocaine dependence. AB - Cocaine was considered incapable of producing dependence in 1980 but was proclaimed the "drug of greatest national public health concern" by 1984. Clinical consensus in 1980 held that cocaine did not produce a withdrawal syndrome, but recent clinical investigations demonstrate that cocaine produces unique abuse and withdrawal patterns that differ from other major abused drugs. Evolving pre-clinical research over the past two decades now suggests that chronic cocaine abuse produces neurophysiological alterations in specific central nervous system systems that regulate the capacity to experience pleasure. These evolving clinical and pre-clinical constructs have led to applications of promising experimental pharmacological treatments for cocaine abuse. PMID- 3043928 TI - [Primary biliary cirrhosis]. PMID- 3043926 TI - Cardiac effects of cocaine: a review. AB - Over the past 15 years, there has been a dramatic increase in the abuse of purified cocaine preparations throughout the industrialized world. The potential lethality of the drug is now recognized, and a growing series of case reports indicate that cardiotoxicity may be an important factor in the morbidity and mortality associated with the drug. Acute myocardial infarction is a demonstrated risk both in subjects with and without pre-existent coronary artery disease. The arrhythmogenic potential of cocaine is less clear and appears to have been overemphasized, although several documented cases of ventricular arrhythmia following cocaine use have been reported. Cardiomyopathy and myocarditis associated with cocaine have also been described, but an etiologic relationship is presently inferential. Based upon presumed pathophysiologic mechanisms, beta adrenergic blocking agents are recommended for arrhythmias, and calcium channel blocking agents and/or nitrates for ischemic syndromes related to cocaine. It is emphasized that these recommendations are based upon a paucity of relevant clinical studies, and controlled clinical trials to establish their efficacy have not been performed. PMID- 3043929 TI - [Current aspects in the treatment of depression]. PMID- 3043930 TI - [100 years of bacteriology--history of the discovery of brucellosis. 1: Uncovering the etiology of Malta fever by the British military surgeon David Bruce and the Mediterranean Fever Commission]. PMID- 3043931 TI - [Possibilities and trends of pharmacotherapy of cardiac arrhythmias in childhood and adolescence. 3: Determinative criteria, therapeutic concept and selection of drugs]. PMID- 3043932 TI - [In memory of Hermann Zondek (1887-1979)]. PMID- 3043933 TI - [Current significance of infectious diseases in childhood]. PMID- 3043934 TI - [Changes in the pathogens causing pneumonia]. PMID- 3043935 TI - [Mechanisms of chronic hepatitis in viral hepatitis B. Hypotheses as rational principles in immune modulating and virostatic therapy of chronic hepatitis B]. PMID- 3043936 TI - [Current epidemiologic, clinical and preventive aspects of hepatitis A]. PMID- 3043937 TI - [Gastrointestinal manifestations of acquired immunodeficiency syndrome (AIDS)]. PMID- 3043938 TI - [Lung diseases in AIDS]. PMID- 3043939 TI - [Is the bone marrow the only source of endogenous odor components? A contribution on immunopsychology]. PMID- 3043941 TI - [Epidemiology and strategy for the control of chronic respiratory diseases]. AB - The conditions of tertiary prevention of chronic bronchitis are determined by prevalence, number of cases of inability to work, time of disease periods and age depending mortality. An age-specific increase could be proved for all parameters. The critical age-line is about the 50th year of age. 75% of the bronchitis patients under medical care are older than 50 years of age. Measure of care ought to be concentrated on the time before the 50th year of age and on cases of risk. By an earlier beginning of medical care it is possible to increase the efficiency of tertiary prevention. PMID- 3043940 TI - [Differential diagnosis of obstruction--clinical aspects of obstructive lung diseases]. AB - The term obstructive lung disease is applied to a group of diseases of different nosology and pathogenesis with the common feature of a characteristic disturbance of the respiratory function consisting in increased resistance of air flow with dyspnoea as the leading clinical symptom. With regard to a differentiated therapy, it is necessary to distinguish precisely between the entities chronic bronchitis, bronchial asthma, emphysema. Meticulous anamnesis, careful clinical examination, basic lung function tests, x-ray examination, electrocardiographic and laboratory examinations, and eventually bronchologic and allergologic methods are sufficient in outpatient and general internal hospital care for a reliable classification and differentiation from a broad spectrum of causes of obstructive symptomatology located outside from the airways. PMID- 3043943 TI - [Patients with chimerism--immunologic detection of bone marrow acceptance]. AB - The establishment of the ABO chimerism (two populations of erythrocytes deviating in ABO antigens) following a bone marrow transplantation represents a simple rapid method for the proof of the transplant acceptance. A remarkable case was described in which in a patient with hypoplasia of the bone marrow in the peripheral blood there exist two deviating in ABO antigens populations of erythrocytes (AB blood corpuscles of the patient and B erythrocytes of the donor) still 4 1/2 years after the transplantation of the bone marrow. This fact speaks for the simultaneous existence of the own bone marrow as well as of the bone marrow of the donor in the patient. PMID- 3043942 TI - [Russia-Halle interrelations in medicine of the 18th century. II. Halle medical delegations in Czar Peter's Russia and later]. AB - In the 18th century for the purpose of amelioration of the medical care by the authorities responsible for the medicinal organization in Russia aimed methods of delegation were brought about. Requests of such kind were sent to the medical faculties of renowned universities. In such requests the University of Halle played a significant role. For the 18th century more than 40 candidates for a Halle doctorate can be authenticated who took Russian services for a certain time or for ever and there obtained important positions during their stay. PMID- 3043944 TI - Effect of omeprazole on ethanol oxidation and aniline hydroxylation in rat hepatic microsomes. AB - Omeprazole, a substituted benzimidazole, is a potent gastric acid antisecretory drug, which inhibits the hepatic oxidative drug metabolism in vitro and in vivo. The effect of omeprazole on the microsomal ethanol oxidizing system (MEOS) and, since ethanol-induced cytochrome P-450 reveals a high activity for aniline hydroxylation, on aniline hydroxylase (AH) has been investigated in rat liver microsomes. Omeprazole inhibits microsomal AH activity significantly in a dose dependent manner, while this was not the case for MEOS activity. These data give indirect evidence that the microsomal metabolism of both ethanol and aniline is mediated by different isoenzymes of cytochrome P-450 and that omeprazole exhibits a different affinity to both compounds. Therefore, it must be emphasized that drug interactions with omeprazole have to be tested experimentally in each individual case, since it is impossible to predict such interactions solely on the knowledge of the drug's metabolic pathway. PMID- 3043946 TI - [Localization of the freezing front in tissue by the ultrasound-A picture technic in cryotherapy]. AB - The knowledge of the depth of infiltration of the freezing front in the treated tissue represented an essential demand for an optimal cryotherapy (sprayfreezing and contactfreezing). The hitherto used invasive measuring methods are connected with a tissue traumatisation and therefore only applicable in a limited way. On that account the non-invasive ultrasound-A-picture-technique was applied for defining the freezing front extension. Its qualification was represented in vitro (gelatin), in living biological tissue (muscle of rabbit), and on the human eye (cryotherapy of the primary glaucoma). The use of the sonography in the cryotherapy can be recommended for other special medical branches too. PMID- 3043945 TI - [ERCP: which contrast medium is suitable?]. AB - To evaluate, wether a new non-ionic contrast medium decreases the complication rate of endoscopic retrograde cholangiopancreaticography (ERCP), we performed a prospective randomized study in 46 indoor patients with suspected pancreatic or bile duct related disease. The low-osmolar low-viscosity non-ionic Iopromid (Ultravist, n = 15), the low-viscosity high-osmolar Ioglicinate (Rayvist, n = 18), and the conventional dissociable high-viscosity Ioxaglinate (Heaxbrix, n = 13), each presenting a iodine content of 300-320 mg/ml were compared. All three contrast solutions gave excellent imaging of pancreatic and bile ducts. No complications, particularly no pancreatitis were observed. Hexabrix caused significant elevations of gamma-GT from 126 U/l to 178 U/l and mof lipase from 144 U/l to 418 U/l (p less than 0.01), respectively. Following Rayvist or Ultravist injections, no significant changes of the leucocytes, SGOT, SGPT, gamma GT, AP, lipase and amylase were observed. We conclude that ERCP performed by skilled investigators is a low risk procedure. Selection of suitable contrast media may diminish hepatotoxic and pancreatotoxic side effects. According to our results, we recommend low-viscosity contrast media (Rayvist, Ultravist). The presumed benefit of the non-ionic solution (Ultravist) could not be demonstrated. PMID- 3043947 TI - [Experimental transplantation of the capsula fibrosa of patients with hepatic echinococcosis to white mice. Analysis of relapses after various surgical procedures]. AB - Systematic histological investigations concerning the existence of protoscolices in the fibrotic capsula and in the liver parenchyma of operated patients were carried out by means of the immuno-fluorescence. A protoscolex in the fibrotic capsula was detectable histologically only once. The theory of Napalkov about the relapses of the disease by protoscolices invaded in the fibrotic capsula and surrounded tissue cannot be corroborated after the transplantation of fibrotic capsula in white mice before and after sterilisation by Scolicid and the other investigations. A method for sterilisation of the fibrotic cavity and the fibrotic capsula is presented. In that way difficult traumatising operations with high lethality like pericystectomy and resection of the liver are avoidable. PMID- 3043948 TI - [Neurologic complications of AIDS]. AB - Clinical symptoms of the central and peripheral nervous system occur in about 40% of patients wit HIV infection. At autopsy, CNS lesions can be demonstrated in even higher percentages. Primary sequelae of HIV infection--either due to direct viral effects or the immunopathologic response of the human host--are acute aseptic meningitis or mengingo-encephalitis, HIV encephalopathy, myelopathy, neuropathy, and myositis. Secondary consequences of immunodeficiency in AIDS are opportunistic infections with other viruses, bacteria, fungi, and protozoa, e.g. CMV, HSV and HZV encephalitis, mycobacterial CNS infections, neurosyphilis, cryptococcal meningitis, and last but not least cerebral toxoplasmosis. The main secondary malignoma of the CNS is lymphoma. Together these disorders form a complex spectrum of central and peripheral neurological symptoms. PMID- 3043950 TI - [Pyoderma gangraenosum with special reference to immunologic and hematologic parameters]. AB - Clinically, the diagnosis "pyoderma gangraenosum" can easily be made on the basis of typical skin features. It has been agreed, so far, that numerous immune deficiencies may play a key role in the pathogenesis of this disease. On reviewing the recent case reports, we observe a change regarding the associated diseases in more than 50%: Whereas earlier reports refer to ulcerative colitis as the most frequent concomitant disease, we now find increasing evidence of hematologic problems in association with pyoderm gangraenosum. PMID- 3043949 TI - [Once daily administration of isoconazole as a cream, solution and spray: comparative studies of patients with dermatomycoses]. AB - In 3 controlled multi-center studies, 372 patients suffering from dermatomycoses were treated with isoconazole as cream, solution, and spray either once or twice daily. Neither with regard to the duration of healing nor the healing rates were observed statistically significant differences between the two modes of treatment. As to the forms of preparation, there were no differences of efficacy either. At the end of treatment, the cure rates amounted to 90-94%. Follow-up examinations carried out two weeks later gave no indications of relapses. PMID- 3043951 TI - [In situ hybridization--an expansion of dermatologic histopathology]. AB - By means of in-situ-hybridization, we are able to detect specifically the mRNAs of defined proteins at a cellular level in frozen sections. This technique not only makes it possible to characterize the biosynthesis of defined proteins at a pretranslational level but can also identify activated cells in their histological surroundings. We employed this technique in order to detect collagen mRNAs in fibrotic diseases of the skin. Using cDNA clones for collagen type I, we proved that in early stages of scleroderma the fibrosis starts in the deep dermis and the subcutaneous fatty tissue. In addition, the regulation of the collagen synthesis seems to be disturbed at a pretranslational level and accompanied by dramatically increased levels of collagen mRNA in fibroblasts. This technique offers objective parameters for collagen mRNA and may be especially useful to estimate the activity of fibrotic diseases in individual patients as well as during therapeutical trials. PMID- 3043952 TI - [Parapsoriasis en plaques. Characterization of the cellular infiltrate using monoclonal antibodies]. AB - We report on a 65-year old woman suffering from parapsoriasis with large plaques. The cellular infiltrate was analysed by means of monoclonal antibodies and an immunoperoxidase technique. Our findings proved that the majority of the cellular infiltrate in the upper dermis is composed of helper-inducer T-cells (Leu 4+, Leu 3a+). A notable number of Leu-2a-reactive suppressor-cytotoxic T-cells associated with hydropic degeneration of the basal cell layer were found within the dermal infiltrate and the dermoepidermal interface. Large numbers of Leu-6-reactive dendritic Langerhans' cells were noted in the lower layers of the epidermis, in particular in areas where the epidermis showed focal exocytosis of lymphoid cells. Langerhans' cells, in contrast, were present in the upper portions of epidermis lacking exocytic T-cells. HLA-DR antigens were expressed on Langerhans' cells as well as on nearly all T-lymphocytes. Positive intercellular staining for HLA-DR antigens were only found in localized areas of the epidermis. Small numbers of both Leu-11B-reactive natural killer cells and cells expressing interleukin-2 receptors were seen within the basal cell layer and at the dermo epidermal junction. Our findings suggest that a cell-mediated immune response which is directed against antigens expressed by keratinocytes may play an important role in the pathogenesis of parapsoriasis en plaques. PMID- 3043953 TI - [Lichen ruber ulcerosus. Differentiation from Graham-Little syndrome. Pathogenesis and therapy]. AB - A 61-year-old female patient having suffered for several years from typical Graham-Little's syndrome developed lichen ulcerosus after a period of 9 years. We discuss the relationship between these clinical manifestations with special reference to the pathogenesis and distribution of skin lesions in lichen rubber ulcerosus. Among the numerous therapeutic approaches, treatment with chloroquine or dapsone as well as skin grafting seem to be most promising. PMID- 3043954 TI - [Circumscribed bullous pemphigoid (Brunsting-Perry) with positive Nikolski phenomenon]. AB - We present a typical localized bullous pemphigoid (Brunsting-Perry) associated with positive Nikolski sign. To the best of our knowledge, this is the first report on positive Nikolski sign in localized bullous pemphidoid. PMID- 3043955 TI - [Serodiagnosis of Lyme borreliosis]. AB - There are various serodiagnostic tests available for the detection of antibodies against Borrelia burgdorferi. An indirect hemagglutination assay, which can detect both IgM and IgG antibodies, was developed for antibody screening. Regarding the confirmation and differentiation of IgM and IgG, we use the indirect immunofluorescence assay (most specific when performed with sera previously absorbed with Treponema phagedenis) as well as the ELISA test. The detection of significantly elevated antibody titers depends on the stage of the disease. In erythema migrans, only 20-50% of the patients are seropositive (with a prevalence of IgM); in neuroborreliosis, the figures amount to 70-90% of the patients (prevalence of IgM in early stages and IgG in advanced stages); and 90 100% of the cases with acrodermatitis and arthritis show elevated titers of IgG (IgM antibodies usually are not detectable in late manifestations). The clinical manifestations of the disease is serologically marked by IgM antibodies or--in neuroborreliosis--by intrathecal production or antibodies which can be detected on account of an increased CSF/serum index. PMID- 3043956 TI - [Recent possibilities for the treatment of osteoporosis in the aged]. AB - Treatment of idiopathic osteoporosis in the elderly presupposes exact radiological diagnosis, the exclusion of a primary illness as the cause of the pathological process and exact differential diagnosis from other metabolic osteopathies. We consider possible means of prevention of the immobilization of old people, and appropriate hormonal substitution in cases of previous illnesses which coincide with a disturbance of the gonadal function, as important prophylactic measures. In the case of manifest osteoporosis, if possible, an assignment of the disease to a manifestation with high or low bone turnover should be made, by means of biochemical adjuvants. In high bone turnover, the substitution of sex hormones or the administration of calcitonin is indicated, particularly if symptoms of pain are distinct. In osteoporosis with low bone turnover, fluoride in long-term therapy is the preferred medication. The latest studies show that a combination of fluoride and active vitamin D metabolites is preferable to monotherapy. All therapy for this disease, independent of the age of the patient, should be supported by isometric exercises, analgesics and appropriate dietary measures. Orthopaedic supporting measures should be applied only if conservative measures in acute vertebral fractures fail. PMID- 3043957 TI - [New clinico-chemical values in chronic joint diseases. Granulocyte elastase and UDP-D-xylose:proteoglycan core protein-beta-D-xylosyltransferase of chondrocytes]. PMID- 3043958 TI - [Enzyme immunoassay for 17-hydroxyprogesterone in serum and saliva and from blood microfilter paper and microtitration plates]. PMID- 3043960 TI - [Experiences with intra-arterial digital subtraction angiography in arteriography of the hand]. PMID- 3043959 TI - [Relation of echogenicity to the angle of incidence on musculature and tendon tissue]. AB - Echogenic structures of the locomotor system are demonstrated with many or few echoes, depending on the angle at which ultrasound waves strike them. This causes a graduation in the demonstration of arcuate tendons in different planes, from dense to sparse echoes (the former in the case of portions perpendicular to the angle of incidence of the ultrasound waves, the latter in the case of portions which the waves strike at a slanting angle). This phenomenon occurs with arcuate tendons and is known as the "deflection phenomenon." It must be taken into account when assessing structural changes in tendons in order to avoid misinterpretations. Similar conditions are found in the muscles. They are important in those groups of muscles which pass over major joints with changed courses or manifest pronounced pinnation with different courses of their septa. In the musculature, failure to take this phenomenon into account causes changes in echogenicity to be misinterpreted as ruptures or partial ruptures. PMID- 3043961 TI - Leriche syndrome; pre-operative assessment using digital subtraction angiography. PMID- 3043962 TI - Arteriomegaly: the value of intravenous digital subtraction angiography. PMID- 3043963 TI - Quality assessment and assurance in primary health care. PMID- 3043964 TI - [Presentation of the document "Model type of a plan for prevention at the canton level," drafted by a study group of the Swiss Society for Social and Preventive Medicine]. AB - This article presents the preliminary work of a group charged to develop a model for a prevention plan on the level of the cantons. The document should advise them as to how to develop a comprehensive prevention plan. This work has been stimulated by the reaction of the Conference of Cantonal Health Ministers to the "Concept 1986" of the Swiss Society for Social and Preventive Medicine. PMID- 3043965 TI - [Expert opinion on biologic stains. Determination of status, future trends]. AB - In this paper an attempt is made to critically review the literature, with special emphasis on bloodstain analysis. One essential aim is the integration of this field into casework. Three basic components in skillful assessment of stains are described: (1) analysis of stain morphology, (2) discriminating and attributing analyses, (3) individualization. Regarding the first, the analysis of stain morphology is based upon the extensive experimental literature published since 1895--mainly in continental Europe. Since 1971 there have also been publications in the American literature. The large family of stain forms and their dependency on multiple variables are described, especially regarding the modes of formation, the energy of impact, and the physical properties of the substrate. The essential elements for reconstruction of the crime are described. The areas of application are arranged in case groups. Since in case work the stain pattern is complicated by many artifacts and overlaps, forensic pathologists are considered the ideal experts for the analysis of bloodstain patterns, as they have a profound knowledge of the type and sequence of injuries. If this is not the case, the forensic pathologist should at least be integrated into the investigating team. In practical application, the stain form is not always adequately analysed. The education and training of pathologists should be improved to achieve this standard. Analysis of the stain morphology and a subsequent selection of stains are also essential prerequisites for meaningful further investigations. By the use of discriminating and attributing analyses, one can as a rule arrive at a definite answer by using only one test. This is true for basic questions such as the identification of blood type, as well as proof of exclusion. One can distinguish between traditional methods, the new field of immunochemistry and rarely used methods. Immunochemistry has permitted success in recent years in determination of the blood group from hair. It is recommended that reference laboratories be established for training in these rare methods. Individualization analyses are subdivided into two large fields: non-DNA individualization and DNA individualization. It is postulated that in the future stain laboratory both areas will coexist. In non-DNA individualization, essential progress has been made. The detection of protein polymorphisms by blotting and subsequent visualization by antibody-linked enzyme/substrate reactions has led to a considerable increase in sensitivity and specificity.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3043966 TI - [Morphologic studies in criminal justice--requirements and reality]. AB - A critical analysis is given of the investigation and examination of fatalities for the public prosecution office and for the courts. Rationalistic simplifications in legal quarters, changes in legal procedures and bureaucracy have had negative effects on the field of forensic medicine. It is questionable whether forensic science today can fulfil the scientific demands with regard to a complete and thorough explanation of the cause of death. The reduced interest in expert opinions concerning the pattern of injuries that can help in the reconstruction of the act or in judging the intention and guilt of the perpetrator is a striking and alarming development in criminal justice. Medical examinations in the somatic field are no longer used enough in forensic cases; with regard to the administration of justice, the role of the forensic expert is limited to the postmortem findings and the cause of death. On the other hand, there is increased interest in the expert opinion of psychologists, psychiatrists and specialists in the field of drugs and alcohol. The fact that the number of autopsies and histological as well as toxicological examinations ordered by the public prosecutor is rather small has serious and negative implications for establishing the truth. Such orders are merely dependent on the circumstance of death and the situation in which the body was found. However, the external circumstances can be misleading or they can be manipulated. That is why some crimes are not revealed. In the Federal Republic of Germany, if there is no suspicion of external violence permission is granted for corpses to be buried without the cause of death being clarified by autopsy. However, the fact that the opinions of forensic specialists differ regarding this development in the administration of justice must also be criticized. There is an increasing tendency for investigations to be carried out only if they are ordered by the prosecutor and for questions to be answered only if they are asked officially. Even our colleagues are influenced too much by external circumstances and consider an autopsy unnecessary if the cause of death seems to be clear (for instance in cases of hanging, drowning or bleeding to death). Second autopsies have shown that the thoroughness and the quality of the first autopsy often leave much to be desired. It is, for instance, unforgivable if organs are described without being examined. Special interest is afforded to the necessity for the significance of histological examinations.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3043968 TI - [One-step urethroplasty--indications, results and surgical peculiarities]. AB - In a representative patient group the long-term results were examined after one stage urethroplasty with a subcutaneous pedicle skin flap. The used modification in which an intact strip of epithelium is buried shows good to satisfactory results in 80% of the cases, but in more than 60% a diverticulum of the neourethra was found. Therefore, in 25% of all urethroplasties a secondary resection of the diverticulum was necessary. First experiences with the use of a skin patch to reconstruct the urethra have shown that formation of diverticula can be prevented by this technique. The pre-, intra- and postoperative measures (epilation, secretion drainage, temporary urinary diversion) which are necessary to achieve good functional results are presented in detail. PMID- 3043967 TI - [Significance of functional and nutritive pulmonary circulation for vital reactions in the form of embolisms]. AB - The functional and nutritive circulation in the lungs is connected by anastomoses between the pulmonary and bronchial arteries. The anastomoses have the structure of blocked arteries from which arteriovenous anastomoses proceed to the peribronchial plexus. The pulmonary artery is provided with a flow impulse by the anastomoses, and oxygen-containing blood is admixed with the venous blood, thus forming an "aortalization" in the lungs. By diverting the bloodstream, venous blood can reach the bronchial artery. The peculiarities of the lung circulation are important for vital reactions in the form of macro- and microembolisms. Macroembolisms prove the functionality of the system if branches of the pulmonary artery are closed before the arteries are blocked. A hemorrhagic infarction either arises or does not arise, and the hemorrhagic infarction cannot exceed a certain limit. A microembolism is over and above the anastomoses. If the microembolism is greater, pressure in the arteria pulmonaris can cause blood from the pulmonary artery to overflow into the bronchial artery. Because arteriovenous anastomoses arise from the blocked arteries, microemboli can now reach the systemic circulation. Thus, the system described can explain the passage of microemboli into the systemic circulation, avoiding the capillaries of the lungs; on the other hand, larger microembolisms can prove the functionality of the system. PMID- 3043969 TI - [Pathogen spectrum of non-gonorrhea urethritis]. AB - By cultural investigations in 333 males with non-gonorrhoical urethritis (NGU) and 158 control probands it was demonstrated that Chlamydia trachomatis is the main germ of this disease also in the GDR with a frequency of 40% of the cases. A chlamydia-negative NGU was caused in 15% by Ureaplasma urealyticum in a number of greater than or equal to 10(4) germs/ml urine and in 5% by Trichomonas vaginalis, respectively. Candida albicans occurred significant more frequently in the patient group (21%) than in the control group (4%). Other optional pathogenic bacterias were the cause of NGU in single cases. The diagnosis and therapy of NGU should be considered this germ spectrum and the sexual partner also. PMID- 3043970 TI - [Our surgical heritage. In memory of Prof. Dr. Wilhelm Muller, Professor Ordinarius for surgery 1901-1928 in Rostock]. PMID- 3043971 TI - [Perioperative risk from the viewpoint of the surgeon. Analysis and clinical use]. AB - General definitions are proposed in this paper for the terms of risk and risk factor. Risks are then analysed, with reference to the literature, on the basis of the defined terminology. Possible approaches are then demonstrated for clinical translation of risk analyses into appropriate therapeutic concepts, with reference being made to a study of oesophageal carcinoma. The benefit has been persuasively reflected in reduced lethality. PMID- 3043974 TI - [History of medicine. Prof. Dr. Walter Hohlweg on his 85th birthday]. PMID- 3043973 TI - [Comparison of the progesterone test and uterus sonography as screening procedures in the detection of patients at risk of endometrial cancer]. AB - The increasing incidence of endometrial cancer requires more attention to early detection. Postmenopausal women without symptoms were examined by progesterone challenge test (96 diabetics, 111 without diabetes) and by sonography (44 diabetics, 74 without diabetes) in order to recognize proliferation of the endometrium. The application of progesterone induced a bleeding in 4% of the women. The diagnostic curettage performed 4-6 weeks after this test revealed almost always atrophic endometrium. We found a good correspondence between abnormalities of the uterine cavity detected by sonography and the results of the pathological examination after notice pathological changes of the endometrium. PMID- 3043972 TI - [Value of colon sonography within the scope of surgical diagnosis]. AB - The sensitivity, specificity, and predictable potential of colon sonography were determined by comparison between sonographic, colonoscopic, radiological, and surgically secured findings. Also compared were sonographic colon findings obtained from unprepared patients (n = 74) with those obtained from patients prepared by means of orthograde mannitol irrigations (n = 53). No major differences were found to exist between both groups. Indications for colon sonography in the context of surgical diagnosis are outlines on the basis of the above results. PMID- 3043975 TI - [Clinical and ultrasonic diagnosis of hematometra following curettage]. AB - By hands of a case report we describe our diagnostic management in a patient suffering from hematometra 3 months after legal abortion. Sonography plays an important role in such cases to detect and explain the symptoms "metrorrhagy and uterine cramps monthly connected with small inflammatory signs". For curing we did a probing of the uterine cavity to eliminate the residual blood coagulas. 3 months after curing the hematometra the patient was clinically and sonographically without bodily ailments. Hypotheses about etiology and treatment of this rare condition are reviewed. PMID- 3043977 TI - [Breast milk jaundice--a harmless symptom or a reason for stopping breast feeding?]. AB - Increased interest in breast-feeding focuses attention on an undesirable side effect--the breast milk jaundice of the newborn. In this review a complication is given on contemporary knowledge with respect to types of breast milk jaundice, etiology, diagnostic and preventive strategies. Especially the early and late onset types of breast milk jaundice are distinguished. Though of the jaundice aggravating effect of some human milks, there is no reason to avoid breast feeding. Improved hospital nursing policies and careful information may help to encourage the mothers to continue in breast-feeding successfully. PMID- 3043976 TI - [Nutrition in pregnancy]. AB - Review about during pregnancy. Nutrition of a pregnant woman has to cover the maternal need of energy and additionally the stuff for the fetus as preconditions for its growth. In the trimester an extra intake of 15 grams protein is necessary, but no one of fat and carbohydrates. An increase of 400-500 mg Calcium, 10 mg fernum and vitamins A, B1, B2 and C is very important. Smoking, alcohol and drug abuse may be injurious to the fetus. Passive smoking of the mothers, therefore automatically of the fetus, too, is harmful for the fetus. Our recommendations for nutrition of pregnant women demonstrate, that feeding during pregnancy is simple. PMID- 3043978 TI - [Non-immunologic fetal ascites--case report]. AB - In this paper 4 cases with fetal nonimmunologic ascites are reported. Etiology from ascites and outcome of pregnancies are discussed. To elucidate the cause of ascites may be important for further pregnancies. PMID- 3043979 TI - [Twin pregnancy with fetal hydrocephalus]. AB - There is a case report on twin pregnancy with fetal hydrocephalus, recognized in the 24th week of gestation. Course of pregnancy and management during labour are described. The peculiarity of our observation is for the constellation mother- healthy fetus--fetus with hydrocephalus. Taking care for the pregnant woman and the healthy fetus are most important. PMID- 3043980 TI - [Congenital hypoplastic left ventricle--a case report]. AB - A case of congenital hypoplasia of left heart ventricle is presented with regard to its clinical and pathophysiological aspects. A detailed prenatal ultrasonic or echocardiographic diagnosis seems to be absolutely necessary. Each sonographer should at least be able to advise of any suspicion of fetal heart abnormalities. PMID- 3043981 TI - Improved computer-assisted reading of identification and shortened MIC data for reporting on urine specimens at a Berlin university hospital. AB - Results of bacteriological analyses of urine specimens from patients of Steglitz University Hospital at Berlin Free University are transmitted directly from the laboratory reading station to a mini-computer by means of an input tablet and a menu-driven programme. First, an identification of each specimen is entered on reception and printed out to form a working list. After culturing, the microorganisms are inoculated into microdilution plates for susceptibility testing and differentiation of Enterobacteriaceae. At the reading station, the urine data acquisition programme stores the observed CFU/ml values as well as up to 6 pathogens for each specimen. For each pathogen susceptibilities to 30 antibiotics are tested and reported for up to 4 levels of sensitivity. Enterobacteriaceae are speciated largely according to the results of 17 biochemical tests. User-programme interaction can be monitored on an LCD. Two correction facilities are provided: (1) within the current specimen, or (2) by recalling earlier specimens. Antibiotic usage patterns and species coding may be modified without reprogramming by editing the configuration files for (1) usage of the menu, (2) species coding, and (3) substrate-species combinations. The results are printed on self-adhesive labels, which are returned to the senders of specimens on the conventional request forms. PMID- 3043982 TI - Recombinant plasmid DNA variation of Clostridium oncolyticum--model experiments of cancerostatic gene transfer. AB - The specific germinating capacity of spores of Clostridium oncolyticum in tumours in vivo, and various reports on concomitant partial oncolysis (tumour lysis) have prompted us to propose a concept of equipping C. oncolyticum with genes of other organisms producing cancerostatics. As an example we used Colicin E3, whose structural genes lies in the E. coli plasmid pCo1E3-CA38. After in vitro recombination of restrictase EcoRI-fragmented pCo1E3-CA38 DNA with an uncharacterized plasmid fraction from C. concolyticum a plasmid-free strain of C. oncolyticum was infected with recombinant DNA. A special microbiological selection system allows the identification of C. oncolyticum clones with Colicin E3-similar formation of cleared haloes. PMID- 3043983 TI - Renal pathology and spleen cell chemiluminescence of mice infected with a wild type and a low-virulence mutant of Candida albicans. AB - Pathogenicity and virulence factors were studied for a wild-type strain of Candida albicans (MY 1044) and an auxotrophic, temperature-sensitive mutant strain (MY 1049) that was derived by ultraviolet irradiation. The mutant was a temperature-sensitive, serine auxotroph. Renal pathology and chemiluminescence of spleen cells from infected mice were assessed in an attempt to identify virulence factors. Renal damage was evident following intravenous infection with either strain, although the mutant appeared to be less invasive; MY 1044 produced characteristic miliary, subcapsular lesions, while the mutant (MY 1049) produced large granulomas. Spleen cells from each infected group were stimulated in vitro with either phytohemagglutinin, concanavalin A or opsonized yeast cells to measure the respiratory burst using a luminol-dependent chemiluminescence assay. The highest chemiluminescence responses correlated with severe renal damage (uremia) and not with yeast virulence. No differences in chemiluminescence were observed among spleen cells from mice infected with either strain when renal pathology was minimal. PMID- 3043984 TI - [Meningitis as a complication of anterior-sacral meningocele]. AB - A 19-year-old young man was admitted with a serious clinical picture of an infected anterior sacral meningocele. Escherichia coli was identified as the pathogenic organism in the spinal putrid fluid. A catheter percutaneously introduced in the hernial sac served as an external drainage of the fluid. At the same time a systemic antibiotic treatment was carried out. After the sanation, the opening of the meningocele was surgically closed in the interval transdurally from the dorsal direction. For this fibrinous adhesive was used in addition to the suture. After six months the hernial sac closure could be demonstrated in the check-up myelo- and computer tomogram. PMID- 3043985 TI - Cerebral abscesses secondary to otorhinolaryngological infections. A study of 386 cases. AB - A study is presented, of adjacent cerebral abscesses secondary to otorhinolaryngological infections. A total of 386 patients were included. According to frequency, and in relation to the adjacent septic focus the cerebral abscesses were classified as: otogenic--334 cases (86.5%); rhinogenic--47 cases (12.2%); tonsillary--5 cases (1.3%). These abscesses had similar characteristics: they occurred with the highest frequency in the second and the third decade of life, had a higher incidence in males, the otorhinolaryngologic infection had a chronic course with frequent recidives, propagation of the infection from the original focus occurred by continuity or through the venous system, and the clinical picture was marked by symptoms of local and general infection, by meningeal manifestation, and frequently by altered consciousness (somnolence, confusion). Surgical treatment, next to other therapeutic measures (administration of antibiotics, anti-inflammatory drugs, and intensive care) was aimed at improving the vital prognosis. The postoperative mortality was 23.8%. It was also aimed at increasing the functional recovery rate. Neuropsychical sequellae were noted in 54.9% of all cases. After surgery for the cerebral abscess attempts were made at removing the septic focus, preventing any additional risk of cerebral seeding. PMID- 3043986 TI - Some ultrasonic values of carotid arteries. AB - Examinations of both common carotid arteries in men and women divided into three age categories (up to 30 years, from 30 to 50 years, over 50 years) were performed by ultrasonographic B-scan and Doppler technique. The average value of the maximal systolic speed of blood flow was in men 100.55 cm.s-1, in women 99.10 cm.s-1. The average speed of blood flow in men as well as women at the age up to 30 years was 110 cm.s-1; at the age from 30 to 50 years 102 cm.s-1 (-7.2%); at the age over 50 years 86 cm.s-1 (-21.8%). The average diameter D of the right common carotid artery in men was 6.37 mm, of the left 7.0 mm; in women 5.75 mm of the right and 6.0 mm of the left. PMID- 3043987 TI - Rapid healing of diabetic foot ulcers with meticulous blood glucose control. AB - Fifteen diabetic patients, with neuropathic food ulcers refractory to conventional treatment, were found to be poorly balanced and were put on meticulous regimens; some on continuous subcutaneous insulin infusion and others on split mixed doses. Once diabetes was controlled, the wound healed rapidly in 11 of the patients within 4 to 13 weeks. In 4 patients amputation was necessary. The outcome was better in patients with good peripheral pulses. We suggest that tight control of diabetes promotes healing of diabetic foot lesions. PMID- 3043989 TI - Clinical experience in pancreas transplantation in Lyon: long-term survival of duct injected pancreatic grafts. AB - Ninety-seven pancreatic grafts in 92 insulin-dependent diabetic patients were performed during the last 11 years. Eighty-three of these grafts were carried out after neoprene duct injection, the other patients underwent pancreato-duodenal transplantation. In 80 cases, a double pancreas and kidney graft was performed. Five different immunosuppressive protocols were subsequently applied. Actuarial survival of patients and pancreata was 75.1% and 47%, after one year and 54.6% and 22.1%, respectively, 4 years after transplantation. Slightly better results were observed in double pancreas and kidney transplantation. The survival of both patients and pancreas improved when the most recent immunosuppressive protocols including cyclosporin A and only small doses of steroids were applied. The main causes of loss of the pancreatic graft were rejection, vascular thrombosis and death of the patient with functioning organ. Metabolic studies showed good insulin secretion with normal or impaired glucose tolerance as well as good short and half-term glycemic control. Whole pancreas grafts with enteric diversion yielded prompter and higher insulin secretion but the incidence of surgical complications was increased. In comparison to the data recorded at 6 months after pancreas transplantation, 5 patients of our series with still functioning organ showed an equally satisfactory and unchanged glycemic control after more than 4 years from surgery. In these patients, the previously high insulinemic values decreased to normal levels. However, 3 of these patients showed a decrease in post-prandial peaks as confirmed also by OGTT. However, mean blood glucose level was not altered. In our series the suppression of exocrine pancreatic secretion by neoprene duct injection did not appear to represent a relevant cause of decrease in endocrine function. The results obtained do not yet allow us to draw definite conclusions as to the efficacy of pancreas transplantation in the treatment of degenerative complications in diabetic patients. PMID- 3043988 TI - Management of diabetics with the use of a microprocessor: comparison of insulin treatments based on blood and urine glucose levels. AB - The insulin treatment of 8 insulin-dependent diabetics was controlled with a microprocessor (Better Control Medical Computer, BCMC, Inc., Toronto, Canada) with information derived from blood or first voided urine glucose concentrations assessed by reagent strips four times a day, before the three main meals and bedtime snack. The microprocessor recommends modification of the insulin doses so as to reach a pre-prandial blood glucose value of 110 mg/dl or a urine glucose concentration of 0.1 g/dl. During the first two weeks self-management was uniformly applied by the patients, based on their blood glucose concentration. Subsequently, it was continued by the patients who were divided into two groups, one using the blood, the other the urine glucose concentrations, each for three weeks, alternately. During microprocessor treatment the patients' mean blood glucose profiles decreased from 152 +/- 37 mg/dl to 126 +/- 28 mg/dl. No difference was found between treatments based on blood or urine glucose concentrations concerning either the mean blood glucose profiles or the number of hypoglycemic episodes in the presence of an average glucose threshold and good renal function. The first voided urine glucose concentration and mean and maximal blood glucose values obtained at the time of urine filtration were closely correlated (r = 0.82 and 0.86, p less than 0.001). PMID- 3043991 TI - [Syncopes in patients of advanced age]. AB - This review covers the various etiologies and therapeutic possibilities of syncopes in the elderly. The importance of normal blood pressure and the danger of hypotension are stressed: The frequently occurring vascular stenoses may become especially dangerous if pre-stenotic pressure drops as a consequence of orthostatic dysregulation or of inadequate treatment of hypertension. Orthostatic dysregulation--a common phenomenon in old age--is possibly due to a diminished activity of the baroreceptor reflexes or to the pooling of blood in the venous system and/or to the diminution of blood volume by therapeutic procedures (diuretics) or by bed rest. The cautious use of antihypertonics, of diuretics and of psychopharmacological drugs in the correctly adjusted dose is essential in the management of these pathophysiological mechanisms in the elderly. The orthostatic hypotension--a condition that has to be treated in the old aged--can be managed by supportive treatment such as education of baroreceptor reflexes, compression of the lower extremities, and correction of electrolyte and water disturbances. Vasoconstrictors have to be used additionally with great care to adjust the correct dose of the combination of dihydroergotamine and sympathomimetics individually. PMID- 3043990 TI - Evaluation of B-cell secretion and peripheral insulin resistance during pregnancy and after delivery in gestational diabetes mellitus with obesity. AB - Nine pregnant women with gestational diabetes mellitus (GDM) were studied. Six normal pregnant women and six normal nonpregnant women were evaluated as control groups. All the women underwent oral glucose tolerance test and glucose clamp during the third trimester of pregnancy and two months after delivery. During OGTT, glucose, C-peptide and insulin plasma levels were determined. C-peptide and insulin values in the late phase of OGTT were higher during pregnancy than after delivery in both groups. In gestational diabetic women, the M-value in the second steady-state during glucose clamp was lower than in controls, both during pregnancy and after delivery. Nevertheless, in both groups the M-value during pregnancy was lower than after delivery. Moreover, in gestational diabetic women there was an inverse correlation between M-value in the second steady-state and ponderal excess index after delivery. In conclusion, the impaired peripheral glucose utilization and the pancreatic pattern of gestational diabetic women compared to normal suggested altered B-cell secretion response, increased peripheral resistance and overweight to be the main changes in GDM. PMID- 3043992 TI - [Follow-up in patients with syncope]. AB - The prognosis of patients with syncopes depends on their age, underlying illness and the available possibilities of treatment. Patients over 70 years and those with syncope caused by cardiovascular disease have a particularly unfavourable prognosis. If the cause of syncope has been clearly diagnosed, 40 to 100% of the patients can be treated effectively. In 40% of all patients with syncope, the cause remains uncertain. In a follow-up period of 2 years, the cause of syncope could be discovered in a further 12% of patients by examinations on an outpatient basis. Of the patients for whom the reason of syncope was unexplained, 75% remained free from syncopes throughout this 2 year observation period, while 8% suffered from further attacks. In some of these patients it is possible to find a probable diagnosis by combining their symptoms. PMID- 3043993 TI - [Instrumental diagnosis of peripheral arterial occlusive disease]. AB - The increasing frequency of peripheral occlusive arterial disease in the general population emphasizes the need of suitable methods for early detection of this disease. This overview summarizes the methods of current use of angiologic investigation. Their sensitivity and practicability are critically discussed. Mechanical and electronic oscillography are simple screening methods and allow a slight localization of arterial occlusive disease. Exercise tests increase the sensitivity of oscillographic methods. Continuous wave Doppler-ultrasound can be used for measuring peripheral arterial pressure, directional or pulsed Doppler ultrasound are needed for the determination of flow direction and for registration of flow-pressure curves. Multichannel pulsed Doppler-ultrasound is needed for quantitative flow registration in determined arteries. Detailed high quality pictures of the arterial structures can be obtained by high resolution B mode scanners-while information on flow properties is given by frequency-analysis of Doppler signals achieved in duplex scans or color-coded scans. Plethysmography is still the standard method for quantitative flow determination in upper and lower extremities, suitable for the evaluation of the vascular capacity of the examined member. Capillaroscopy gives a direct view in the situation of microcirculation, which is very valuable for the management of diabetic microangiopathy and collagen diseases. Thermography offers the opportunity of registration of impressive imaging of functional vascular reactions. PMID- 3043994 TI - A double-blind placebo-controlled trial of indobufen in the prophylaxis of migraine. AB - In a double-blind randomized trial of 42 patients with classic or common migraine, indobufen, an antiplatelet drug that inhibits platelet cyclo-oxygenase, was compared with placebo in the prevention of migraine. The duration of treatment was 3 months, and the efficacy was assessed on the basis of the following variables: frequency and duration of attacks, headache index (intensity x frequency); 35 completed the investigation. Indobufen at an oral dose of 200 mg b.i.d. reduced all the variables considered, where placebo did not. The drug was generally well tolerated. The findings of the preliminary trial suggest that indobufen might be an useful alternative in the prophylaxis of migraine. PMID- 3043995 TI - Amaurosis fugax: clinical, Doppler and angiographic findings. AB - Clinical, Doppler and angiographic findings are described in 53 consecutive patients who presented with amaurosis fugax (AF) in a total of 57 eyes; 4 had non simultaneous attacks in both eyes. Atherosclerotic lesions were detected on Doppler or angiographic (conventional arteriography and/or intravenous digital subtraction angiography) examination in 36 (63%) of the relevant precerebral internal carotid arteries (ICA) in 34 patients. Nineteen (53%) of these lesions caused a diameter reduction of more than 75%. Patient age was the most important factor in predicting the presence of relevant carotid occlusive disease, all 36 lesions being found in patients over 50 years of age. Two unusual cases of AF are described; in one AF was caused by stenosis of the ipsilateral ophthalmic artery, and in another by occlusion of the brachiocephalic artery with a steal syndrome from the right common carotid artery (CCA) to the right subclavian artery. PMID- 3043996 TI - Descriptive epidemiology of Creutzfeldt-Jakob disease in Finland. AB - In 1974-84 30 patients died with a diagnosis of Creutzfeldt-Jakob disease (CJD) in Finland. Sixteen of the patients were pathologically confirmed and 14 were probable cases; 6 were familial (20%). One further familial patient was alive at the end of 1984. The median age of the 30 patients at death was 59.5 years (range 46-73 years). The familial patients were significantly younger than sporadic cases (median and range 49.5 and 46-57 versus 61.5 and 51-73; P less than 0.01). Only 5 of the 24 sporadic patients were men (male to female ratio 1:3.8). The annual number of new cases as well as the death rate increased in the late 1970's (annual death rate 0.57 per million in 1974-84 and 0.91 in 1979-84). This probably reflects the growing awareness of CJD among neurologists, rather than a real increase of the incidence and death rate. Annual age-specific death rate per 1 million population in 1974-84 reached a peak value of 2.57 in the age group of 60-64 years. In sporadic CJD the age-specific death rate of women was higher than that of men in all age groups. A chronic medical condition, precedent or concomitant with CJD, was seen in 15 patients. PMID- 3043997 TI - Risk factors and genetic background for Alzheimer's disease. AB - Analytic epidemiology has contributed significantly to the generation and testing of hypotheses of the causes of AD. Several case-control studies have indicated risk factors related to the genetic hypothesis, such as: the presence of cases of either AD or Down's syndrome in other family members and the advanced age of the mother at subject's birth. In this respect recent molecular biology studies on DNA from patients affected by the familial form of AD, have demonstrated a genetic polymorphism localized on chromosome 21. On the same chromosome, the gene coding for beta-amyloid has been also recently localized. Immune, viral and toxic factors, thought to cause AD, have been also investigated in case-control studies, none of them has been found consistently associated with the disease, with the only exception of the head trauma. On the other hand most case-control studies have been carried out in younger cases and no large studies are yet available for late onset patients. PMID- 3043999 TI - HIV infection in the Nordic countries. PMID- 3044000 TI - Methods and results of epilepsy surgery. PMID- 3043998 TI - Potential pharmacotherapy of Alzheimer disease. A comparison of various forms of physostigmine administration. AB - This paper reviews clinical trials with physostigmine administered to Alzheimer patients using three different routes of administration: oral, i.v. and intracerebroventricular (i.c.v.). It compares results obtained with three different routes by the authors as well as by other authors. Particular emphasis is given to a novel type of physostigmine administration, the i.c.v. route. Advantages and disadvantages, as well as side effects of each route are presented and discussed. PMID- 3044001 TI - Occlusal contact wear of prosthodontic materials. An in vivo study. AB - The wear of gold, porcelain, and heat- and light-cured resins in occlusal contact with resin and porcelain teeth has been studied in vivo. Both weighing of removable segments of fixed partial prostheses and a replica technique allowing scanning electron microscope observations of the worn surfaces were used. The results showed that all materials had the greatest loss of substance when the opposing teeth were of porcelain. The heat-cured, unfilled resin was the least wear-resistant material, followed by light-cured resin, porcelain, and gold. The heat-cured resin showed a combined tribochemical and fatigue type of wear. The light-cured resin and porcelain showed mainly a fatigue type of wear, whereas gold showed a combined abrasive and fatigue type of wear. PMID- 3044002 TI - [Cytologic exam with fine needle aspiration in neoplastic pathology of the parathyroid gland]. PMID- 3044004 TI - New aspects of the theory of microcirculation. AB - New aspects and details of the mechanism of the microcirculation in the tissues are discussed in the paper. Particular attention is payed to the role of the blood pressure amplitude for the filtration and resorption processes through the capillary wall. A new model of the microcirculation is discribed in which the interstitial space is considered as a functional unit determining the intensity of the transcapillary exchange processes. The components of the hydraulic interstitial pressure are characterized as well as the process of formation and transport of limphatic fluid. The proposed model is a base for the better understanding of the theory of microcirculation. PMID- 3044003 TI - [Malignant neoplasms of the minor salivary glands]. PMID- 3044005 TI - Psychiatric Munchausen's syndrome. Literature review with case reports. AB - The present study reviews the literature on Munchausen's syndrome. In a psychiatric hospital, out of a total of 775 admissions under the age of 65 in 1986, four patients were diagnosed as having Munchausen's syndrome (0.5%). The underlying pathology of these cases is discussed and suggestions for management and further research made. PMID- 3044006 TI - Post-diction of suicide in a group of depressive patients. AB - Prediction of suicides was made on the basis of 99 psychiatric patients included in a study of anti-depressive therapy during 1961-63. These patients were followed up until 31 December 1984. Eight of them committed suicide. The prediction was based on two test methods, the Serial Colour-Word Test (CWT), which registers style of adaptation to a conflicting situation, and the Meta Contrast Technique (MCT), where incongruous or threatening stimulation is introduced by degrees into neutral pictures and various defense misrepresentations of the threat are spotted. Both tests could predict suicide, but MCT was most successful. The sensitivity of the test was 0.80 and the specificity 0.75. The main risk symptom appeared to be depressive retardation together with lack of functional defensive structures. PMID- 3044007 TI - Sedative effects of maprotiline and amitriptyline. AB - By means of critical flicker fusion measurements and subjective estimation of drowsiness, experiments with 12 healthy subjects over periods of 8 h showed that maprotiline caused less than half the amount of sedation compared to amitriptyline, both drugs given in single doses of 50 mg. The experiments were blind, randomized and placebo controlled. Implications for clinical use are discussed. PMID- 3044008 TI - Thyroid hormones in the treatment of affective disorders. AB - The experience of using thyroid hormones in affective disorders is summarized. This includes: 1) Using thyroid hormones alone in depression; 2) their combined use with tricyclic antidepressants; 3) addition of thyroid hormones to nontricyclic antidepressants; 4) the use of thyroid stimulating hormone; and 5) thyrotropin releasing hormone in depression. Suggested mechanisms of action are discussed. A special attention is paid to the place of thyroid hormones in the treatment of rapid cycling affective disorder. PMID- 3044009 TI - Locus ceruleus morphometry in aging and schizophrenia. AB - The locus ceruleus (LC), a pigmented brainstem nucleus rich in noradrenergic neurons, has been proposed to be involved in the pathophysiology of aging and schizophrenia. We undertook a quantitative neuropathological study of the LC in these two conditions. A computing planimeter was employed to count the total number of neurons and measure the volume of the LC, neuronal cross-sectional area, and percent of neuronal area occupied by neuromelanin in the brains of 39 subjects; 13 "normative", 15 leucotomized schizophrenic (most had died in the preneuroleptic era), and 11 leucotomized non-schizophrenic control patients, ranging in age from 11 to 94 years. There was a significant inverse correlation between age and total number of LC neurons, neuronal size, and LC volume, and a significant positive correlation between age and the percentage of neuronal area occupied by neuromelanin. Although schizophrenics did not differ significantly from control groups on any of the parameters of LC morphology, there was a trend for reduced LC volume in schizophrenic brains. Also, the LC of leucotomized patients tended to have increased neuromelanin content and slightly increased cell counts compared to normals, although the importance of this finding is not clear. PMID- 3044010 TI - Investigations of the prevalence of psychiatric disorders. AB - More than thirty studies of the prevalence of psychiatric disorders have been reported from Western countries in the last thirty years. Significant methodological advances have occurred, including sophistication of sampling techniques, improvements in caseness definitions and identification of cases, and clear definitions and specification of rules for diagnosis. The introduction of standardized diagnostic instruments for use by trained lay interviewers is expected to facilitate studies of psychiatric epidemiology. The methods used in each of the studies are briefly described. PMID- 3044011 TI - Boys who became offenders. A follow-up study of 2203 boys tested with projective methods. AB - Future offenders have more ratings than controls on variables indicating psychopathology. The characteristics which most consistently and significantly discriminate between preoffenders and controls, and which also turned out to be the best predictors of future criminality, are those which previously have been observed in subjects with serious mental disorders. The results indicate that the high degree of psychopathology in subsequent offenders is associated with distortion of body image and with poor reality orientation, i.e. that the most prominent characteristics of future offenders are related to a weakness of the ego. Future offenders seem to have a sufficiently developed ego to handle common tasks. However, their ego fails when they are confronted with unfamiliar and frustrating situations. The inability of subsequent offenders to complete a task properly, with failure on difficult and provocative tasks, shows that they have not developed adequate defence mechanisms and that they have a low tolerance for frustration. They are emotionally immature, instable and impulsive. In short, they are dominated by id impulses. There is no indication of gender confusion (homosexuality) in subsequent offenders. However, there are marked differences between the drawings by cases and by controls regarding characteristics related to sexuality. These findings are interpreted as an exaggeration of masculinity and an abnormal preoccupation with sexuality combined with anxiety for the opposite sex. It is possible that severe criminality, contrary to milder forms of lawbreaking, is associated with elevated self-esteem and extroversion. There are indications that ultimate offenders experience human beings (others or themselves) as deviant, monstrous and inhuman, features which indicate that distorted self-concept and distorted self-perception are characteristics of preoffenders. The findings regarding aggression are inconsistent, just as they were in prior projects. The Draw-A-Person Test (DAP) and Colour Slide Test (CST) findings indicate that preoffenders are more aggressive and destructive than controls, whereas the Hand Test (HT) shows neither more aggression nor more acting-out tendencies in future offenders than in controls. The literature offers conflicting findings regarding depression. The CST results indicate that depression is more common in preoffenders than in controls.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3044012 TI - International Symposium on GnRH Analogues in Cancer and Human Reproduction. Geneva, Switzerland, 18-21 February 1988. Abstracts. PMID- 3044014 TI - Symbolic trepanations of skulls from the Middle Ages (IXth-Xth century) in Bulgaria. AB - Symbolic trepanations (ST) of skulls (IXth-Xth century) found in north-eastern Bulgaria near the West coast of the Black Sea were Studied. It was established that trepanations were performed on subjects well over the age of thirty and cover 41% from the total material which is the highest frequency documented so far. Tracks of the ST are localized most often on and round the sagittal suture, bregma and lambda. Their number ranges from 1 to 9 per cranium with a diameter of 3-4 to 20-28 mm and a depth of 1 to 6 mm. They are usually round and encircled with a prominent ridge. In this article we try to explain the reasons for the symbolic trepanations as well as the link between the traditions of the ancient Turkish nations and Chinese medicine. PMID- 3044013 TI - Cerebral angiography under stereotactic conditions. Technical note. AB - A simple system for cerebral angiography under stereotactic conditions using standard components is described. X-ray tubes and power generator belong to the usual equipment of the stereotactic operating room. A film changer and an injection pump were integrated into the stereotactic arrangement. Angiographic documents with reduced X-ray magnification were obtained. For topographical orientation 5 films per seriogram were sufficient. The system also allows stereoscopic viewing by performing an additional oblique sequence after the head ring is rotated exactly 6 degrees. The combination of the necessary diagnostic procedures, as CT scan, angiography, ventriculography, serial biopsy within a single three coordinate reference system offers considerable advantages for brain tumour patients. PMID- 3044016 TI - Suicide and the rural adolescent. AB - The rural family, with its particular stressors, is increasingly vulnerable to overwhelming crises. Adolescent suicide, although rare, may result from or add to that stress. The intent of this study was to identify and examine specific stressors with which a rural adolescent must deal, the coping mechanisms utilized, and when these fail, how it leads to suicidal behavior. Therapeutic modalities available to the troubled adolescent were examined and additional services suggested. PMID- 3044015 TI - The cranial nerves as living morphotic entities--Friedrich Arnold's Icones nervorum capitis, Heidelberg 1834. AB - Friedrich Arnold's neuroanatomical treatise Icones nervorum capitis published in its first edition in Heidelberg 1834 ranks scientifically and iconographically among the most brilliant works of 19th century anatomical literature. Consonant with Arnold's conceptions the lithographed engravings depict the cranial nerves as living, morphotic entities comprising their topographical origin and periphery in a distinctness and beauty never been seen before. PMID- 3044017 TI - Adolescent suicide: character traits of high-risk teenagers. AB - Adolescent suicide rates are increasing and are currently higher than ever recorded. Thus, it has become essential for health professionals, counselors, and parents to become familiar with characteristics of the high-risk teenager. This article examines personality traits and life circumstances which place an adolescent at higher risk for suicide. Among the variables examined are: depression, acute suicidal behavior, poor family relationships, alcohol and drug use, recent loss, failure in school, and other characteristics. As adolescents pass through difficult life stages successfully, teenage suicides will decrease. Professional helpers and parents must be able to recognize the signs which are discussed and take an active role in prevention and/or intervention. PMID- 3044018 TI - Optical anisotropy decay studies of the dynamic structure of myosin filaments. AB - We applied flash-induced absorption and phosphorescence anisotropy decay methods to the study of rotational motions of myosin heads in solution), myofibrils and muscle fibers); myosin heads were selectively labeled with a triplet probe EMI (5 eosinylmaleimide). EMI-labeled subfragment 1 (S1) showed a single exponential decay of anisotropy over two decades; the analysis indicated that if S1 is modeled as a prolate ellipsoid of revolution, the major axis was 16-17 nm and the minor axis 4.7-4.5 nm. The decay curve of myosin filaments could be simulated by double-exponentials-plus-constant approximation. The data could be analyzed by a double-cone model), in which we assumed that a head part (S1), wobbles in the first cone and a rod portion next to the head also wobbles in the second cone. The semiangle of each cone was estimated to be 35 and 48 degrees, respectively. We found that myosin heads in myofibrils under relaxing conditions extensively rotated as in myosin filaments in solution. When the spacing between thick and thin filaments was artificially reduced by the increase of osmotic pressure with the addition of polyvinylpyrrolidone (PVP), restriction of the angular range of the rotational motion was observed. Under rigor conditions no motion was observed in a 10 microsecond time scale, indicating that the heads were immobilized by binding to thin filaments. Preliminary results on the rotational motions of myosin heads in muscle fibers are also reported. PMID- 3044020 TI - Assessment and management of the cancer patient with depression. PMID- 3044019 TI - Helix-coil melting in rigor and activated cross-bridges of skeletal muscle. AB - The studies described in this paper focus on the structural stability of the S-2 segment of the myosin cross-bridge in rigor, relaxed and activated muscle. Enzyme probe observations of myofibrils of rabbit psoas muscle in rigor reveal that the alpha-helical LMM/HMM hinge domain of S-2 undergoes substantial local melting near physiological temperatures when the S-2 portion of the cross-bridge is detached from the thick filament surface. This process is strongly suppressed under ionic conditions where the cross-bridge is bound to the filament backbone. Activation of glycerinated fiber bundles results in a dramatic increase (approximately 100 fold compared to rigor and relaxed fibers) in the rate of chymotryptic cleavage in the hinge domain consistent with an increase in local melting at several sites encompassing this region. Comparative plots of the apparent rate-constant for cleavage within the S-2 hinge and the isometric force generated by active fibers versus [MgATP] give similar profiles suggesting a close coupling between this conformational transition and contractile force. This interpretation appears to be in accord with recent laser T-jump experiments of rigor ("bridges up") and activated psoas muscle fibers which also suggest coupling between melting in S-2 and force generation. PMID- 3044021 TI - The role of concrete services in cancer care. PMID- 3044022 TI - The effects of cancer therapies on the central nervous system. PMID- 3044023 TI - Cancer's impact on caregivers. PMID- 3044024 TI - Neuropsychiatric evaluation and treatment of delirium in cancer patients. PMID- 3044025 TI - Understanding denial in cancer patients. PMID- 3044026 TI - Psychotherapy with cancer patients. PMID- 3044027 TI - The application of behavior therapy in oncology. PMID- 3044028 TI - Family issues in cancer care. PMID- 3044029 TI - [Pediatrics in Berlin and its historical contribution to health care of children and adolescents]. PMID- 3044030 TI - Enhanced antibacterial resistance in neutropenic mice treated with human recombinant interleukin-1 beta. AB - Human Recombinant IL-1 was investigated for its ability to increase non-specific resistance to Staphylococcus aureus in neutropenic mice. Mice, rendered neutropenic with cyclophosphamide and then administered IL-1 intraperitoneally, demonstrated enhanced resistance to subsequent challenge with S. aureus as measured by increased survival and bacterial clearance. No protective effects were observed with heat inactivated IL-1. Efficacy was observed only when both IL 1 and the subsequent bacterial challenge were administered into the same site. Despite the observed protective effects, animals treated with IL-1 did not have increased numbers of blood leukocytes or peritoneal phagocytes prior to infection or at the times coincident with bacterial clearance. Based upon these observations, enhanced activity of resident macrophages may be responsible for the protective effects observed in IL-1 treated mice. PMID- 3044031 TI - Action of a novel drug (Zy 16039) on mucus secretion in the ferret isolated trachea in vitro. AB - 1. The effect of 4-H-2-carboxamido-4-phenyl-thieno-[3,2c]-[1]-benzopyran (Zy 16039) was examined on the smooth muscle contraction, mucus secretion and albumin transudation in the ferret whole trachea in vitro. 2. Zy 16039 (0.1-20 microM) produced a concentration-dependent relaxation of the ferret trachea contracted by methacholine (1 microM) and phenylephrine (10 microM). The relaxations were about 20% of the full contractions. 3. Zy 16039 has no effect on the resting (zero) output of mucus in the ferret trachea. Methacholine-induced mucus secretion was significantly inhibited by Zy 16039, whereas phenylephrine-induced secretion was significantly increased. 4. Methacholine-induced secretion of lysozyme, a marker of serous cell secretion, was inhibited by Zy 16039 both with regard to output and concentration of lysozyme. In contrast, Zy 16039 significantly increased the output of lysozyme due to phenylephrine, with no effect on concentration. 5. Zy 16039 had no significant effect on the rate of output of fluorescent albumin through the tracheal wall. However the concentration of albumin in the mucus samples was changed because of the effect of Zy 16039 on mucus secretion induced by methacholine and phenylephrine. 6. We conclude that Zy 16039 relaxes airway smooth muscle, and either promotes or inhibits mucus secretion depending on its source. It has qualitatively similar actions to vasoactive intestinal peptide. PMID- 3044032 TI - [Changes in the human corneal endothelial cell junction in vivo]. PMID- 3044033 TI - Barrett's esophagus: a radiologic diagnosis? PMID- 3044034 TI - Duplex sonography of hepatic artery thrombosis after liver transplantation. AB - When hepatic artery thrombosis occurs after liver transplantation, another transplantation is required to ensure the patient's survival. Because of the importance of establishing this diagnosis, we reviewed the results of duplex sonography in 37 angiographically or surgically proved cases of hepatic artery thrombosis after liver transplantation. There were 20 children and 17 adults. Ten of the 20 children had angiographically documented hepatopetal arterial collaterals. Such collaterals were not seen in the adult patients. This subset of patients was evaluated separately to determine if intrahepatic arterial blood flow reestablished by collaterals after hepatic artery thrombosis was a cause of false-negative Doppler studies. Thirty-four (92%) of the 37 cases of hepatic artery thrombosis were correctly identified by Doppler. A Doppler pulse was not identified in any of the children with arterial collaterals. We conclude that duplex sonography is sensitive in detecting hepatic artery thrombosis after liver transplantation. Furthermore, the presence of blood flow in hepatopetal arterial collaterals does not cause false-negative examinations. PMID- 3044035 TI - CT- or sonography-guided biopsy of the liver in the presence of ascites: frequency of complications. AB - The presence of ascites has been considered a contraindication to percutaneous biopsy of the liver. To determine the validity of this assumption, we performed percutaneous biopsies of the liver under CT or sonographic guidance in 28 patients who had ascites and in 28 patients who did not have ascites and compared the complication rates in the two groups. Twenty-two patients (79%) in the group with ascites and 19 patients (68%) in the group without ascites had biopsies to determine the cause or extent of chronic liver disease. The remainder were oncologic patients who had biopsies to determine the cause of a focal hepatic mass. The complication rate in the patients who had ascites (32%) was less than that in the patients who did not have ascites (43%) (the difference did not reach statistical significance, p less than .30). In the ascites group, complications included transient hypotension (five patients), a mild-to-moderate fall in hematocrit (three patients), and a small leak of ascites from the biopsy site (one patient). In the control group, minor complications included transient hypotension (three patients), a mild-to-moderate fall in hematocrit (seven patients), and a small subcapsular hematoma (one patient). One major complication occurred in the control group: a patient required a blood transfusion because of the fall in his hematocrit. We conclude that the complication rate in liver biopsies guided by CT or sonography in the presence of ascites is not higher than similar biopsies done in the absence of ascites. Ascites should not be considered a contraindication for performing such biopsies. PMID- 3044036 TI - CT diagnosis of renal angiomyolipoma: the importance of detecting small amounts of fat. AB - Six patients were reviewed who had renal angiomyolipoma (1.2-4.0 cm) in which only minimal amounts of fat were evident on CT. The fat content of the lesion was appreciated because tissue attenuation measurements of small areas of low attenuation within the tumors were performed and because thin-section (5-mm) and nonenhanced CT scans were used. The fat content of the lesions could be identified on 10-mm sections in three cases but only on 5-mm sections in three others. In two cases, fat was seen only on the nonenhanced 5-mm thin sections. Careful sampling of low-density regions within the mass must be performed because a single region of interest over the entire tumor will produce an average attenuation in the soft-tissue range. The use of 5-mm thin sections and thin, nonenhanced CT sections increases spatial and density resolution and decreases susceptibility to partial-volume effects. In a correlative study, no areas of fat were detected in a review of 100 well-circumscribed (4.0 cm or smaller) renal cell carcinomas. Detecting the existence of fat in a renal lesion will establish the diagnosis of angiomyolipoma and is the only radiologic finding that can differentiate it from renal cell carcinoma. Thus, unnecessary surgery will be avoided in these cases. PMID- 3044037 TI - Fat-filled postoperative renal cortical defects: sonographic and CT appearance. AB - Surgical filling of renal cortical wedge resection defects with vascularized retroperitoneal fat resulted in postoperative sonographic and CT appearances that simulated focal renal masses in four patients. Correct identification of this abnormality is important in order to avoid unnecessary further evaluation to exclude renal neoplasm. PMID- 3044039 TI - Urine leaks after renal transplantation: value of percutaneous pyelography and drainage for diagnosis and treatment. AB - We reviewed our experience with 12 renal transplant patients who had urine leaks to compare the accuracies of sonography and nuclear renography with that of antegrade pyelography in establishing the diagnosis. The leak was proved by surgery in 11 of the 12 cases. We also determined the role of diverting percutaneous nephrostomy drainage in the treatment of such leaks. The diagnosis was established by sonography in eight (67%) of the 12 patients. Nuclear renography, performed in nine patients, showed decreased renal function but showed the leak in only three (33%) of the nine cases. Antegrade pyelography, performed in all 12 patients, showed leakage in 10 (83%). In the other two patients, follow-up nephrostograms done within 24 hr showed leaks near the ureterovesical anastomotic site. Seven of 11 patients who were managed with a combination of percutaneous nephrostomy drainage and surgical reconstruction were treated successfully (i.e., a functioning graft was retained); however, only one patient was managed successfully by percutaneous methods alone. Antegrade pyelography is more accurate than sonography and nuclear renography in the detection of urine leakage after renal transplantation. However, percutaneous nephrostomy drainage appears useful only as an adjunct to surgery for treatment of this complication. PMID- 3044038 TI - Urinary obstruction in renal transplants: diagnosis by antegrade pyelography and results of percutaneous treatment. AB - We reviewed our experience with 51 renal transplants to evaluate the accuracy of antegrade pyelography as compared with that of sonography and nuclear renography in the diagnosis of transplant obstruction. Also, the results of percutaneous treatment were analyzed in 44 of these patients. Obstruction was clinically suspected in all of the patients (increased serum creatinine levels and decreased urine output). Antegrade pyelography showed obstruction in 44 (86%) of the 51 patients, and nephrostomy drainage catheters were inserted. Sonography showed pyelocaliectasis in all 49 cases in which it was performed; in 42, the pyelocaliectasis was due to obstruction (14% false-positive rate). Nuclear renography showed obstruction in only six (18%) of 33 cases in which it was performed; all six cases proved to be obstructed (0% false-positive rate and 82% false-negative rate). Twenty-two (50%) of the 44 patients treated with nephrostomy drainage were managed successfully without surgical intervention; seven of these 22 required balloon dilation of ureteric strictures in addition to catheter decompression of the collecting system. The average duration of catheterization required for successful percutaneous treatment was 35 days. This experience suggests that antegrade pyelography has a definite role in the workup of patients suspected of having renal transplant obstruction. The percutaneous access permits successful catheter drainage. Compared with antegrade pyelography, sonography is reasonably accurate in determining the presence of urinary obstruction, although false-positive diagnoses are found in a substantial number of patients. Nuclear renography is not, however, a useful indicator of obstruction owing to its high false-negative rate. Percutaneous treatment of urinary obstruction in transplantation patients proves successful in approximately 50% of cases. PMID- 3044040 TI - Evaluation of lopamidol and diatrizoate in excretory urography: a double-blind clinical study. AB - Image opacification, patients' tolerance, and clinical and laboratory findings were evaluated in patients having excretory urography at three centers. In a double-blind, parallel study, iopamidol was compared with diatrizoate (50-ml dose), and in an open-label trial, the administration of a 100-ml dose of iopamidol was evaluated. In the double-blind study, a total of 84 patients received 50 ml of either iopamidol or diatrizoate. In the open-label study, another 42 patients received a 100-ml dose of iopamidol. Image opacification scores after the administration of the 50-ml doses showed better opacification with iopamidol than with diatrizoate in the renal calices (p less than .05) and in the composite kidney (p less than .05). Opacification scores were higher for 100-ml doses of iopamidol than for 50-ml doses in all anatomic regions as well as in the composite kidney (p = .0001). Patients' tolerance to iopamidol was significantly better than their tolerance to diatrizoate (p less than .025). Investigators observed adverse drug reactions in a total of 10 patients. In the double-blind study, one of 43 patients had transient bradycardia after the administration of iopamidol. In the same study, four of 41 patients who received diatrizoate had five minor adverse drug reactions. With 100-ml doses of iopamidol, five of 42 patients had adverse reactions. No adverse side effects required therapy in either study. There were no significant changes in vital signs or laboratory values after drug administration. The results of this study show that iopamidol is a suitable agent for excretory urography at doses of 50 and 100 ml. Patients report fewer unpleasant side effects with iopamidol than with diatrizoate. Overall image quality was better with iopamidol than with diatrizoate. Overall evaluation of drug performance was better with iopamidol than with diatrizoate. PMID- 3044041 TI - Intramural Teflon injection of the ureter for treatment of vesicoureteral reflux: sonographic appearance. AB - Endoscopic injection of Teflon paste under the submucosal portion of the ureter in the bladder is a new treatment for vesicoureteral reflux. Twenty-one children at The Montreal Children's Hospital and at St. Justine Hospital were treated with this technique over a 2-year period. In 15 children, abdominal sonography was performed 1-5 days after Teflon paste injections. Follow-up sonograms performed 5 weeks to 1 year after the procedure are available in nine children. Sonographically, the Teflon paste at the injection site appears as a hyperechoic focus within the bladder wall with distal shadowing seen postoperatively and on follow-up examinations. This finding occurred in 22 (88%) of 25 treated ureters. The location of Teflon paste after injection as viewed via cystoscopy is correlated with the sonographic appearance. This preliminary report suggests that sonography is useful in determining the location and size of the Teflon mass, in evaluating surrounding soft tissues at various time intervals after injection, and in assessing possible complications such as obstruction. PMID- 3044042 TI - Prenatal diagnosis of anencephaly: spectrum of sonographic appearances and distinction from the amniotic band syndrome. AB - To document the characteristic sonographic abnormalities of anencephaly and to identify potentially confusing sonographic features, we reviewed 20 cases of anencephaly. All of these cases were diagnosed prenatally with sonography after 14 menstrual weeks in patients who were seen at our institution between 1984 and 1988. In all cases, the correct diagnosis was made on the prenatal sonograms and was confirmed pathologically. The sonographic diagnosis was primarily based on the absence of brain and calvarium superior to the orbits on coronal views of the fetal head. This typical appearance was altered by the presence of echogenic tissue superior to the orbits in nine (45%) of 20 cases. Pathologically, the tissue corresponded to angiomatous stroma (area cerebrovasculosa) and appeared quite sizable on sonograms in four fetuses (20%). It may appear solid or mixed solid and cystic. In one fetus, it appeared brainlike. Despite this appearance, the sonologist should not be dissuaded from the diagnosis of classic anencephaly. Hydramnios occurred in seven (35%) of 20 patients, and oligohydramnios occurred in none of the patients. Anencephaly may be distinguished from the cranial defects associated with the amniotic band syndrome (amputation defects that occur as the sequelae of amniotic disruption) on the basis of the symmetry of the cranial defects (100% of anencephalic fetuses in this series) and the absence of limb, body wall, and spinal abnormalities that typically accompany the amniotic band syndrome. Although there may be minor variations in the sonographic appearance of the cranial defect of anencephalic fetuses (i.e., much or little angiomatous stroma), we conclude that this anomaly can be accurately detected and diagnosed on fetal sonograms obtained after 14 weeks menstrual age and distinguished from the amniotic band syndrome. PMID- 3044043 TI - Nonspecificity of the "to-and-fro" sign vs the "whirlwind" sign specific to pseudoaneurysms. PMID- 3044045 TI - Pneumomediastinum and pneumopericardium after cocaine abuse. PMID- 3044044 TI - The effervescent gallbladder. PMID- 3044046 TI - Small intestine enema. PMID- 3044047 TI - Faculty development: meeting the needs of postsecondary educators of deaf students. PMID- 3044048 TI - The CHARGE association: implications for teachers. PMID- 3044049 TI - Malaria: chemoprophylaxis and therapy. AB - Malaria should be considered in a patient with unexplained fever and a history of travel to an endemic area. Aggressive therapy must be started if Plasmodium falciparum infection is a possibility. Travelers must be educated about mosquito bite protection and appropriate chemoprophylaxis. Travelers can, however, acquire malaria despite chemoprophylaxis, and symptoms may appear up to one year after the trip. PMID- 3044051 TI - Pressure support ventilation. AB - The management of patients with respiratory failure can be challenging. A relatively new technique may be useful in freeing these patients from mechanical ventilation. With a spontaneously taken breath, positive pressure is provided by a ventilator equipped to detect small changes in airway pressure. Pressure support ventilation may mean the difference between an independent life and ventilator dependency. PMID- 3044050 TI - Sarcoidosis. AB - The classic chest radiograph, showing bilateral hilar and right paratracheal adenopathy, is found in half of sarcoidosis patients. Pulmonary infiltrates are a major cause of morbidity and mortality. Extrapulmonary disease may be found in the skin, eyes, liver and heart, and in the nervous, musculoskeletal and other systems. Despite myriad pathologic and biochemical abnormalities, the typical patient is asymptomatic. Most patients with symptomatic sarcoidosis benefit from steroid therapy; in some, however, the disease progresses inexorably. PMID- 3044052 TI - Avoidable nondietary risk factors for cancer. AB - Smoking cessation has the greatest potential for reducing cancer mortality. Moderate physical activity may prevent colon cancer by modifying bowel peristalsis patterns and may prevent breast cancer by modifying ovulatory frequency. Radiation therapy and the use of certain drugs may increase the risk of some cancers. Liver cancer may be the most preventable major cancer. PMID- 3044053 TI - Tic disorders of childhood. AB - Tic disorders may be classified as simple tics, chronic motor tics or Tourette syndrome, the most severe of the three types. Tourette syndrome is rather uncommon and is often misdiagnosed. Tic disorders of childhood probably have an organic etiology and a genetic component. An increased incidence of obsessive compulsive behavior, attention deficit disorder and other abnormalities have been reported in Tourette patients. Haloperidol is often useful in controlling severe tic symptoms, while stimulant drugs may worsen the symptoms. PMID- 3044054 TI - Avulsion fractures involving the cruciate ligaments. AB - The evaluation of a child with an acute knee injury is difficult. The injury may be an avulsion fracture involving either of the cruciate ligaments; a common cause is a cycling accident. The amount of fragment displacement dictates the management. Minimal displacement can be treated with immobilization in flexion. Larger displacement requires internal fixation, which can be accomplished arthroscopically. Rehabilitation must be early and aggressive. PMID- 3044055 TI - Supernumerary nipples. AB - Supernumerary nipples are common anomalies, and their significance is usually limited to cosmetic concerns. However, a high index of suspicion should be maintained during physical examinations, because any disease process that involves anatomically normal breasts may affect aberrantly located breasts or nipples as well. These anomalies may be associated with several systemic disorders, particularly urinary tract abnormalities. PMID- 3044056 TI - Amphotericin B nephrotoxicity. AB - Nephrotoxicity becomes apparent days to months after the institution of amphotericin B therapy. It is characterized by azotemia, decreased renal plasma flow, decreased glomerular filtration rate, tubular defects, nephrocalcinosis, and diffuse, nonspecific histologic changes. Management consists of minimizing exposure to other nephrotoxins and ensuring adequate hydration. PMID- 3044057 TI - Prostaglandin E2 induction of labor. PMID- 3044058 TI - ACE inhibitors as initial therapy for hypertension [correction]. PMID- 3044059 TI - Current therapeutic principles in the acute management of severe congestive heart failure. AB - The phrase "heart failure" encompasses many clinical entities. The therapeutic principles determining the treatment of these entities vary according to the etiology of congestive heart failure (CHF), the existing hemodynamics, and the mode of action of different drugs. In acute CHF due to myocardial ischemia, intracellular acidosis and the accumulation of phosphate may be the initial underlying causes of contractile failure while, minutes later, lack of high energy compounds may be an important contributory factor. The cause of contractile failure in chronic syndromes is less well understood. There is evidence for the desensitization of beta receptors on the cell surface but the precise location of the defect is unclear. The receptors may be down-regulated but, in addition, abnormalities have been reported in several other parts of the contractile pathway including the contractile proteins and the sarcoplasmic reticulum. Deficiency of cyclic adenosine monophosphate has also been suggested as a mechanism of contractile failure. In both acute and chronic CHF, there is a redistribution of blood flow to the body organs. Of particular significance is the reduction of blood flow to the kidneys, and a reversal of this defect is one of the major therapeutic objectives. Positive inotropic drugs, vasodilators and drugs altering relaxation of the heart, have been evaluated in the treatment of CHF. Pure inotropic drugs can cause tachycardia, ischemia and "metabolic exhaustion" of the myocardium. The most advantageous profile for an "inotropic" drug in many patients with CHF would be a drug combining systemic vasodilatation, renal vasodilatation, increased relaxation of the myocardium only a mild positive inotropic effect and no chronotropic effect. PMID- 3044060 TI - Clinical development of dopexamine hydrochloride (Dopacard) and an overview of its hemodynamic effects. AB - A clinical development program was initiated to identify the hemodynamic profile of activity of dopexamine hydrochloride. Studies in healthy subjects confirmed the cardiovascular activity of dopexamine hydrochloride and demonstrated its potency. Doses of 0.5 to 8 micrograms/kg/min doubled cardiac output without change in mean blood pressure, although pulse pressure did widen. In patients with stable chronic cardiac failure (mainly New York Heart Association class III), dopexamine hydrochloride (4 micrograms/kg/min) increased stroke volume 47 +/- 9%, cardiac index 64 +/- 10% and heart rate 11.7 +/- 3%. Systemic vascular resistance decreased by 42 +/- 5%. At higher doses, cardiac index increased further, but chronotropic effects became more prominent. Subsequent studies in patients recovering from cardiac surgery and studies of longer duration in patients with chronic congestive heart failure have demonstrated a similar hemodynamic profile that is sustained throughout the infusion. The renal vasodilator effects have been confirmed in both healthy subjects and patients. With all catecholamines the balance of chronotropic to inotropic or vasodilator effect is critical. Dopexamine hydrochloride (1 to 4 micrograms/kg/min) increases cardiac index by over 40% at clinically insignificant (10%) increases of heart rate. PMID- 3044061 TI - Mechanisms of supraventricular tachycardia. AB - Programmed electrical stimulation of the heart in combination with intracardiac recordings has contributed a wealth of new information on the mechanisms and pathways of supraventricular tachycardia in humans. This knowledge has resulted in better treatment approaches to these patients. Questions still remain, however, about the mechanisms of atrial flutter and fibrillation and of some types of atrial tachycardia. The location of the circuit in paroxysmal atrioventricular nodal tachycardia also continues to puzzle investigators. The use of refined mapping techniques during cardiac surgery should provide answers to these questions in the near future. PMID- 3044062 TI - Resting metabolic rate and postprandial thermogenesis in vegetarians and nonvegetarians. AB - Resting metabolic rate (RMR), thermic effect of a meal (TEM), and associated hormonal changes were studied in vegetarians and nonvegetarians. RMR was established by indirect calorimetry in 12 male vegetarians (VEG) and 11 nonvegetarians (NVEG) of similar body fat and aerobic fitness. Subjects ingested a liquid meal and TEM was measured for 180 min postprandially. Plasma concentrations of glucose, insulin and thyroid hormones (T3 and T4) were determined before and after meal ingestion. Absolute RMR was comparable between VEG and NVEG. However, TEM was lower (p less than 0.01) in VEG (55.8 +/- 3.3 kcal/180 min) vs NVEG (76.4 +/- 3.6). Plasma levels of glucose and insulin were similar between the two groups whereas plasma T3 was slightly but nonsignificantly lower in vegetarians. A vegetarian diet may decrease the postprandial thermic response; this does not support the supposition that an elevated TEM is a factor contributing to the lower body weight in vegetarians than in omnivores. PMID- 3044063 TI - Coagulation defects in cyclosporine A treated allogeneic bone marrow transplant patients. AB - CSA toxicity includes renal impairment, microangiopathic hemolytic anemia (MAHA), thrombocytopenia (T), and consumptive coagulopathy (CC). We report five BMT patients who developed CSA-associated hematological toxicity. All were conditioned with Ara-C, Cyclophosphamide, Methylprednisolone, TBI, and in two cases busulfan. IV CSA was started the day after marrow infusion and, when practicable, changed to the enteral route. Five patients developed MAHA and T resulting in significantly increased transfusion requirements. All patients had renal impairment and red cell fragmentation. In all patients fragmentation was noted before renal impairment. All developed disproportionate increases in BUN relative to serum creatinine consistent with decreased renal perfusion. Hypertension followed renal impairment in four cases and occurred at the same time as the renal impairment in one case. Two developed CC, prolongation in APTT, and marked decreases in plasma fibrinogen. All patients improved on reduction of the CSA dose. BMT recipients receiving CSA at variable doses may develop evidence of a TTP-like syndrome and/or CC. PMID- 3044064 TI - Medical-legal note: Clarence Borel revisited. PMID- 3044065 TI - Asbestos exposure and gastrointestinal malignancy review and meta-analysis. AB - The epidemiologic literature linking asbestos exposure with gastrointestinal malignancy is reviewed. Problems in comparing studies are discussed, appropriate strategies for comparison are developed, and study results are pooled using a model which accounts for both intrastudy and interstudy variability. Stratification of cohorts by dose reveals that significant asbestos exposure, as indicated by a lung cancer standardized mortality ratio (SMR) of at least 200, is associated with an elevated gastrointestinal cancer of SMR for five of six points examined [corrected]. PMID- 3044066 TI - Preventing percutaneous absorption of industrial chemicals: the "skin" denotation. AB - Percutaneous absorption has received comparatively little attention in occupational health, although this route of entry has repeatedly caused occupation-related intoxications. In practice, the evaluation of skin penetration rates is far from simple. Much evidence has been obtained from studies of chemicals used for cosmetics and topical therapeutics, but the information available on compounds encountered in occupational health is limited. The data obtained from experimental studies have confirmed that the concentration, type of vehicle, skin area, skin condition, and extent of occlusion are important factors in determining the degree of percutaneous absorption, but no general model has been developed. Also, too little is known about the basic chemical properties governing the rate of penetration. Thus, prediction is difficult and bound to be rather inaccurate. Current preventive practice follows the procedure used by ACGIH and is mainly based on a "skin" denotation in official listings of chemicals to which exposure limits have been allocated. The number of substances and groups of chemicals which have received skin denotation in 17 selected countries varies between 24 and 179 and a total of 275 are listed as a skin hazard in one or more countries; ACGIH lists 143. Thus, the denotation practice varies. As an unfortunate result of these discrepancies and the dichotomy of skin denotation, the absence of skin denotation may erroneously indicate that efforts to protect the skin are unnecessary. Thus, an evaluation of skin penetration potentials should be incorporated in occupational health practice as a supplement to the official denotations. PMID- 3044067 TI - Clinical benefits and mechanisms of a sustained response to intermittent insulin therapy in type 2 diabetic patients with secondary drug failure. AB - To test the hypothesis that short-term insulin therapy may induce long-lasting metabolic improvements in patients with type 2 diabetes resistant to oral therapy, 19 patients were studied before and four weeks after insulin therapy, and again four weeks after resumption of oral medication. The mechanisms associated with changes of glycemic control after discontinuation of insulin therapy were also evaluated. During insulin therapy, blood glucose levels (228 +/ 13 versus 123 +/- 18 mg/dl, p less than 0.001) and the basal glucose production rate (p less than 0.001) decreased, and the insulin secretory response to glucagon at a standardized glucose level, insulin action in vivo, and insulin binding and action in vitro in fat cells improved significantly. During the post insulin oral therapy, blood glucose levels increased (194 +/- 11 mg/dl, p less than 0.001) but remained below pre-insulin treatment values (p less than 0.01). The mean daily glucose concentration after post-insulin oral therapy correlated with the initial pre-insulin therapy glucose concentration (r = 0.83, p less than 0.001). The improved rate of in vivo glucose disposal and the enhanced insulin secretory response persisted during oral therapy whereas the basal glucose production rate returned to its pre-insulin therapy value. It is concluded that patients with type 2 diabetes in whom oral therapy fails show favorable responses to insulin therapy. After discontinuation of insulin therapy, blood glucose concentrations tend to return to their individual initial values. Therefore, most of these patients require long-term insulin therapy. The mechanism behind the change of glycemic control after cessation of insulin therapy seems to be an increase in the basal glucose production rate rather than deterioration of extrahepatic insulin action or the insulin secretory response. PMID- 3044069 TI - Kaposi's sarcoma: the most common tumor after renal transplantation in Saudi Arabia. AB - Between September 1975 and November 1986, 263 renal transplant recipients at the King Faisal Specialist Hospital and Research Center were followed; 82 procedures were done by the authors using live related donors. Among the 263 patients, 14 cases of Kaposi's sarcoma were identified, an incidence of 5.3 percent compared with an incidence of 0.4 percent in renal transplant recipients from Western countries. In addition, two more patients had other types of tumors. Thus, Kaposi's sarcoma represents 87.5 percent of tumors in the King Faisal Hospital renal transplant population, in contrast to 3.7 percent in the Cincinnati Transplant Tumor Registry. The mean period between transplantation and diagnosis of Kaposi's sarcoma was 12.5 months (range, one to 37 months). Eleven patients were Saudis and three were other Arab nationals. Seven of the 11 Saudi patients were from the southwestern region of the country. Cytomegalovirus titers were not elevated in six of 10 patients. Results of tests for human immunodeficiency virus were negative in seven of eight patients. HLA-A2 antigen frequency was significantly increased in the King Faisal Hospital renal transplant patients with Kaposi's sarcoma as compared with a control population (83.3 percent versus 43.6 percent, p value = 0.006 [P = 0.06 with Bonferroni adjustment]), and increased, though nonsignificantly, compared with the live related kidney transplant recipients without Kaposi's sarcoma (83.3 percent versus 49.4 percent, p value = 0.058 [P = 0.58 with Bonferroni adjustment]), suggesting a genetic predisposition to Kaposi's sarcoma in these patients. PMID- 3044070 TI - Evaluating individualized medical decision analysis. PMID- 3044068 TI - Large variations of sucrose in constant carbohydrate diets in type II diabetes. AB - Several studies show that sucrose does not aggravate hyperglycemia in type II diabetes mellitus, but sucrose is still restricted in dietary recommendations. Since sucrose in high carbohydrate diets elevates fasting triglyceride levels, the effects of sucrose were evaluated in diets with fixed carbohydrate levels. Eighteen diabetic volunteers receiving no medication were given weight maintenance diets with 50 percent carbohydrate, 35 percent fat, 15 percent protein, and 120 g of sucrose for 10 days as inpatients. They were then randomly assigned diets of similar composition with either 220 g of sucrose (high sucrose diet) or less than 3 g of sucrose daily (complex carbohydrate [CHO] diet) for one additional month. There were no differences in fasting, one-, two-, and three hour post-lunch serum glucose levels; 24-hour glycosuria; glycohemoglobin levels; fasting and postprandial serum triglyceride levels, or fasting high-density lipoprotein-cholesterol levels. Twelve patients with preexisting higher triglyceridemia had similar trends, but the postprandial triglyceride levels were lower in the high sucrose diet group of this subset (p less than 0.05 in the third week). Postprandial serum insulin levels declined in the second week on the complex CHO diet. More than 75-fold difference in sucrose intake with constant carbohydrate and fat did not affect glycemic or triglyceridemic control in type II diabetic patients. The reported high sucrose-carbohydrate-induced rise in fasting triglyceridemia was not present when a diet high in sucrose was given without changing total carbohydrate. PMID- 3044071 TI - A review of the ethical and legal aspects of terminating medical care. PMID- 3044072 TI - De novo focal glomerulosclerosis after kidney transplantation. AB - A transplanted kidney in a patient developed focal and segmental glomerulosclerosis, associated with severe systemic hypertension, proteinuria, progressive azotemia, and allograft hypertrophy. A pediatric kidney with two main arteries was used, and occlusion of the artery supplying the upper pole resulted in infarction of this portion of the allograft. Because other known factors predisposing to focal sclerosis were absent, it is postulated that renal hemodynamic changes associated with reduction in functioning renal mass, attended by striking stimuli for renal hypertrophy, resulted in progressive damage. The implications of these concepts are discussed in relation to the progression of renal diseases. PMID- 3044073 TI - Autoantibodies to the insulin receptor as a cause of autoimmune hypoglycemia in systemic lupus erythematosus. AB - A 52-year-old black woman presented with clinical features of systemic lupus erythematosus (SLE) and severe fasting hypoglycemia. Hypoglycemia was secondary to autoantibodies to the insulin receptor that were detected in the patient's serum. There were no anti-insulin antibodies, and other causes of hypoglycemia were excluded. Treatment with high-dose glucocorticoids resulted in restoration of euglycemia associated with resolution of circulating anti-receptor antibodies and parallel improvement in clinical and laboratory features of SLE. This case is compared with other cases of autoimmune hypoglycemia due to anti-receptor antibodies. PMID- 3044074 TI - Anti-thymocyte globulin overdosage--risk or potential benefit? PMID- 3044075 TI - Episodic twice-daily treatment for recurrent genital herpes. AB - A new dosage regimen of orally administered acyclovir, 800 mg twice daily for five days, for the treatment of recurrent genital herpes was compared with the standard dosage of 200 mg given five times per day. A double-blind study of 157 patients was used to evaluate the safety and efficacy of both regimens. The new regimen was well tolerated, more convenient, and as effective as the standard dosage. In male patients, the new regimen may be more effective than the standard regimen of 200 mg given five times per day to treat lesions that are already present. PMID- 3044076 TI - Prolonged continuous versus intermittent oral acyclovir treatment in normal adults with frequently recurring genital herpes simplex virus infection. AB - In Year 1 of this two-year trial, patients with six or more genital herpes recurrences in the past year received suppressive treatment with either acyclovir, 400 mg, or placebo, orally twice daily for one year, and physician documented recurrences were treated with open-labeled acyclovir, 200 mg, orally five times per day for five days (acute treatment). In Year 2, patients received open-labeled acyclovir treatment either with daily suppressive therapy or intermittent acute therapy. Among 683 patients who completed two years of treatment, 348 received continuous suppressive treatment for two years, 276 received acute treatment in Year 1 and suppressive treatment in Year 2, 24 received suppressive treatment in Year 1 and acute treatment in Year 2, and 35 received acute treatment for two years. Patient groups receiving intermittent acute acyclovir treatment experienced means of 7.0 to 12.6 recurrences/year during treatment as compared with 1.4 to 1.9 recurrences/year among groups receiving continuous suppressive treatment. No patients who received acute treatment remained recurrence-free for two years as compared with 29 percent of patients receiving continuous acyclovir suppression for two years. There was no evidence of cumulative toxicity detected by clinical, hematologic, or blood chemistry evaluations performed monthly in Year 1 and quarterly in Year 2. Suppressive oral acyclovir therapy remained effective and well-tolerated in this two-year trial. PMID- 3044078 TI - Natural history and therapy of chronic hepatitis B virus infection. AB - The realization that spontaneous loss of viral replication in chronic hepatitis B virus carriers was associated with remission, by whatever criteria, led directly to the search for agents or combinations of agents capable of abbreviating the duration of viral replication. Of those assessed to date, only alpha-interferons have gained wide acceptance as single-agent therapy. Treatment must be for three months on an alternate-day basis; on this regimen, at least one third of patients will develop clinical and biochemical evidence of an exacerbation of hepatitis followed by rapid clearance from serum of all markers of viral replication, and in some cases, also of hepatitis B surface antigen. A large proportion of carriers remains insensitive to alpha-interferons, however, and studies based on the combination of interferons with other agents that have proved less effective, but with different modes of action, are in progress and appear promising. PMID- 3044079 TI - Treatment of chronic type B hepatitis in Southeast Asia. AB - In Southeast Asia, 15 to 20 percent of the population are hepatitis B surface antigen carriers. The majority of these carriers have chronic hepatitis and would progress to cirrhosis or hepatocellular carcinoma at an annual incidence of 2 percent and 1 percent, respectively. Previous studies from Southeast Asia suggested that immunosuppressive therapy could be harmful, or at best of no value, and antiviral treatment with vidarabine, picibanil, or even interferon was also unsatisfactory. Currently, a randomized controlled trial of human lymphoblastoid interferon, with or without prednisolone pretreatment, versus placebo in patients with hepatitis B core antigen in the liver and hepatitis B e antigen in the serum is being conducted. Forty-five patients (29 receiving interferon, 16 receiving placebo) have been entered in the trial for at least two months. Actuarial analysis shows that the response to interferon therapy was better than that to placebo. Although flu-like symptoms, hair loss, and body weight loss were seen, no side effect requiring specific treatment has been encountered. These preliminary results suggest that human lymphoblastoid interferon is effective and safe in Oriental patients. PMID- 3044077 TI - Human lymphoblastoid interferon for the treatment of chronic hepatitis B. A randomized controlled trial. AB - The aim of this study was to demonstrate whether interferon alone could affect the course of chronic hepatitis B. A total of 66 patients have so far been randomly assigned to receive six months of interferon therapy or no therapy; three patients in the interferon group and two in the control group have withdrawn from the study. Loss of hepatitis B virus-DNA and hepatitis B e antigen was significantly higher in the patients receiving interferon than in the control group; a significant loss of hepatitis B surface antigen was observed only in patients who received interferon. The time to alanine aminotransferase normalization was significantly shorter in patients receiving interferon than in the control group. These data encourage the use of interferon in treatment of chronic hepatitis B. PMID- 3044080 TI - Long-term follow-up of antiviral combination therapy in chronic hepatitis B. AB - For patients with chronic hepatitis B e (HBe)-positive hepatitis, long-term results of pilot studies with lymphoblastoid interferon-alpha, acyclovir, or a combination, and of a randomized controlled trial of interferon/desciclovir combination therapy are presented. HBe seroconversion was observed in more than 40 percent of patients treated with combination therapy, 30 percent with interferon therapy, 18 percent with acyclovir, and 0 percent with no treatment. HBe reactivation occurred in two patients with cirrhosis. Hepatitis B surface seroconversion followed HBe seroconversion in 11 to 30 percent of treated patients. HBe seroconversion was significantly related to initial low levels of viral replication and to transient aminotransferase elevation during the second half of the interferon treatment of 16 weeks. Clinical improvement and persistent normalization of aspartate aminotransferase was observed in all patients with HBe seroconversion. Conversion to a state of virus latency (HBe negative) mostly occurred after therapy, suggesting that the specific immunologic host response had been brought about by the suppression of virus replication through antiviral agents. Recommendations for selection of patients for antiviral combination therapy are made on the basis of these long-term results. PMID- 3044081 TI - Interferons in the treatment of genital human papillomavirus infections. AB - Three major classes of interferons have been identified (alpha, beta, and gamma). All three have been tested in clinical trials in condylomata acuminata, or genital warts, with positive results. Administration by topical, intralesional, intramuscular, and subcutaneous routes results in regression of human papillomavirus genital disease. Significant reduction in measurable lesions occurs in some patients within days of initiation of therapy. Responses appear to be both time and dose dependent. Although disease resolution is highly variable from patient to patient, approximately 75 to 80 percent of all persons show clear clinical benefit at low doses. Biologic side effects of interferons are tolerated well at these doses and occur following systemic or local administration of interferon. In general, the interferons are emerging as active and safe therapeutic agents for genital human papillomavirus infections. This study reviews in detail the series of clinical trials conducted with one of these agents, interferon alpha n1. Results of four small and two major controlled trials in refractory genital warts have proved that this interferon provides significant clinical benefit for the majority of subjects with severe disease. Current studies show that it can be combined safely and effectively with other conventional treatment modalities, such as laser or podophyllin. PMID- 3044082 TI - Spectrum of antiviral activity and mechanism of action of zidovudine. An overview. AB - Zidovudine is a potent in vitro inhibitor of human immunodeficiency virus (HIV) with varying efficacy against other retroviruses. With the exception of Epstein Barr virus, all non-retroviruses tested so far have been insensitive to inhibition by zidovudine. In vivo, efficacy of zidovudine was demonstrated against Rauscher murine leukemia virus and feline leukemia virus. In both experimental models, infections completely resolved in animals when the drug was administered soon after infection. These results suggest that prompt initiation of zidovudine therapy, following a known exposure to HIV, should be considered. Mechanism studies show that zidovudine is phosphorylated to the monophosphate and diphosphate derivatives by the host cell cytosolic thymidine kinase and thymidylate kinase, respectively. The identity of the enzyme that phosphorylates zidovudine diphosphate is not known, but is believed to be the cellular nucleoside diphosphate kinase. The triphosphate of zidovudine appears to be the active form of the drug. Zidovudine triphosphate competes well with thymidine 5' triphosphate for binding to the HIV reverse transcriptase and also functions as an alternative substrate. Incorporation of zidovudine monophosphate results in chain termination. However, it is not clear which mechanism, chain termination or competition with thymidine 5'-triphosphate, or a combination of both, is responsible for the inhibition of HIV replication. PMID- 3044083 TI - Antiviral drug development for the treatment of human immunodeficiency virus infections. An overview. AB - During 1987, a new era in the approach to treatment of severe human immunodeficiency virus (HIV)-I infections was entered. The increased availability of zidovudine and its licensure in many countries have resulted in its widespread use. With these developments have come a multitude of new questions. How long will clinical benefit be sustained? Will zidovudine be useful in asymptomatic HIV carriers and in other HIV-related conditions? How can the diversion of zidovudine to unproven indications be averted so that properly controlled clinical trials are conducted? Can better treatment regimens be developed to enhance the therapeutic/toxic ratio of zidovudine? What are the proper roles of governments in assuring patient access to zidovudine? How should newer anti-HIV drugs be compared with zidovudine? Will combinations of zidovudine and other active agents be useful? To address many of these issues in the United States, the Federal Government has established an AIDS Clinical Trials Group (ACTG) program. Over 3,000 patients have been enrolled in AIDS Clinical Trials Group trials, which span the spectrum from asymptomatic to advanced HIV-I infections. In addition, drug development and screening programs have been established to evaluate new compounds and regimens. Although control of AIDS will be a long and arduous journey, an important beginning has been made. PMID- 3044084 TI - Preclinical toxicology of zidovudine. An overview. AB - The toxicologic potential of zidovudine (azidothymidine) has been extensively investigated in several species. In rats and mice, the median lethal dose was greater than 750 mg/kg intravenously and greater than 3,000 mg/kg orally. In subacute intravenous toxicity studies, no significant toxicologic alterations were seen in rats or dogs. In cynomolgus monkeys, which as in humans rapidly and extensively glucuronidate zidovudine, a reversible, dose-related, macrocytic anemia was seen in animals given 35, 100, or 300 mg/kg per day for three or six months. In three-month and six-month oral toxicity studies in rats, treatment related alterations consisted of a mild increase in glucose level in the blood in female rats in both studies and a reversible, slight-to-mild macrocytic anemia in the six-month study. There was no evidence of teratogenicity in rats or rabbits given the drug during gestation. Results for zidovudine were negative in a bacterial mutagenicity assay, but the drug was weakly mutagenic at concentrations of 1,000 to 5,000 micrograms/ml in mammalian cells. Zidovudine caused chromosomal aberrations in cultured human lymphocytes at concentrations of 3 micrograms/ml and higher and had positive results in a cell transformation assay at concentrations of 0.5 micrograms/ml and higher. No bone marrow chromosomal alterations were noted in a cytogenetics study in rats given zidovudine at several intravenous dose levels up to 300 mg/kg. PMID- 3044085 TI - Acquired immune deficiency syndrome in children. Current problems and therapeutic considerations. AB - Acquired immune deficiency syndrome (AIDS) in children has until recently been under-reported, since the initial Centers for Disease Control definition of AIDS was restrictive. The case definition has now been revised. Most children with AIDS acquired their infection perinatally and have a parent with established AIDS related complex or AIDS or belong to a high-risk group. Prior to March 1985, children also acquired human immunodeficiency virus from a contaminated blood product transfusion or from factor replacement for hemophilia. In the United States, AIDS in children occurs predominantly in cities with large populations of intravenous drug users. There are a number of differences between the clinical manifestations of human immunodeficiency virus infection in children compared with adults. For example, recurrent bacterial infection is more common in children, perhaps reflecting the abnormal B cell function that occurs relatively early in the disease course. Certain opportunistic infections (e.g., toxoplasmosis, cryptococcal meningitis) are less common in children than adults. Lymphocytic interstitial pneumonia does not occur in adults but is found in 30 to 50 percent of children. On the other hand, Kaposi's sarcoma and other malignancies are less common in children. Treatment has consisted largely of general supportive care in hospital or at home; this is dependent on the availability and utilization of resources and financial support. However, as anti retroviral therapy becomes available, studies in children have been initiated. It is hoped that in the future it may be possible to prevent the disease; in the meantime, earlier diagnosis and better therapy are important goals. PMID- 3044087 TI - Results of continued monitoring of participants in the placebo-controlled trial of zidovudine for serious human immunodeficiency virus infection. AB - Zidovudine (AZT, 3'-azido-3'-deoxythymidine) was shown in a controlled trial to decrease the incidence of mortality, reduce the frequency of opportunistic infections, and provide other clinical benefits to patients with acquired immune deficiency syndrome (AIDS) and advanced AIDS-related complex. Two hundred twenty nine patients who participated in the double-blind placebo-controlled trial of zidovudine were enrolled in an open-label study. As of August 31, 1987, and interim analysis indicated that patients receiving zidovudine continued to derive benefits from therapy. Survival rates of zidovudine-treated patients were higher than those that might have been expected from previous experience with similar patients. Patients continued to experience episodes of opportunistic infections; however, these infections were either of decreased severity or were more responsive to conventional therapy. The increase in median CD4 cell counts that occurred initially was followed by a gradual decline to near baseline values. Hematologic toxicities continued to be the major laboratory abnormality associated with drug administration; however, new or more frequent toxicity was not observed with more prolonged therapy. Progressive bone marrow suppression did not appear to be associated with prolonged administration. Overall, patients originally enrolled in the double-blind trial continued to receive clinical benefit from zidovudine therapy. PMID- 3044086 TI - Double-blind, placebo-controlled trial comparing long-term suppressive with short term oral acyclovir therapy for management of recurrent genital herpes. AB - A total of 156 patients with frequently recurring genital herpes were enrolled in a randomized, double-blind, one-year trial comparing long-term suppressive and intermittent oral acyclovir therapy with placebo in the management of recurrent genital herpes. Subjects received either suppressive treatment with acyclovir, 400 mg twice daily for one year, and 200 mg five times per day for five days, for short-term treatment of recurrences; intermittent treatment with placebo, twice daily for one year, and oral acyclovir, 200 mg five times per day for five days, for short-term treatment of recurrences; or treatment with placebo, twice daily for one year, and five times per day for five days for short-term treatment of recurrences. Median time to first recurrence was 250 days for the suppressive group, 28 days for the intermittent group, and 23 days for the placebo group (p = 0.001). The median number of days per month with active disease was 0.32 for the suppressive group, 4.18 for the intermittent group, and 4.72 for the placebo group (p less than 0.001), with a median recurrence rate per 30-day treatment period of 0.08 recurrences/patient in the suppressive group, 0.70 in the intermittent group, and 0.74 in the placebo group (p less than 0.001). Median duration of recurrence was 5.0 days in the suppressive group compared with 6.0 days in the combined intermittent acyclovir plus placebo group (p = 0.001), and was reduced from 7.0 to 6.0 days when intermittent acyclovir treatment was compared with placebo treatment (p = 0.05). Daily administration of oral acyclovir for one year is superior to intermittent or placebo treatment in the management of patients with frequently recurring genital herpes. PMID- 3044088 TI - Long-term suppression with oral acyclovir of recurrent herpes simplex virus infections in otherwise healthy patients. A European multicenter study. AB - In a multicenter study, the efficacy of oral acyclovir 800 mg daily in preventing the development of recurrent herpes simplex virus lesions in 379 patients was assessed. Medication given in two or four divided doses daily for one year was highly effective and well tolerated. A high therapeutic index of acyclovir was maintained throughout the period of study. PMID- 3044090 TI - Management of non-genital herpes simplex virus infections in immunocompetent patients. AB - Non-genital herpes simplex virus in immunocompetent hosts causes a variety of primary infections--gingivostomatitis, keratoconjunctivitis, herpetic whitlow, and encephalomyelitis. Recurrent infections with orolabialis are very common, but are usually mild and self-limiting. Cutaneous complications of herpes simplex virus infections include eczema herpeticum and erytherma multiforme. Systemic treatment with acyclovir is indicated in encephalomyelitis, progressive eczema herpeticum, and frequent severe erythema multiforme. Chronic, suppressive acyclovir treatment may be helpful in severe recurrent infections or those complicated by erythema multiforme/dissemination. Many primary and recurrent infections can be treated with simple topical therapy to control secondary infection. There is no evidence that systemic treatment affects viral latency or recurrent infections following discontinuation of treatment. PMID- 3044089 TI - Suppression of recurrent genital herpes by single daily dosages of acyclovir. AB - Patients with frequent recurrences of genital herpes were treated with oral acyclovir tablets, 800 mg, or placebo once daily for two years. Confirmed recurrences for all patients were treated with acyclovir capsules, 200 mg orally five times per day, for five days. Of 46 patients enrolled, 18 of 22 acyclovir recipients and 14 of 24 placebo recipients completed two years of study. Patients receiving acyclovir experienced a mean of 0.184 recurrences/month compared with a mean of 0.977 recurrences/month for patients receiving placebo (p less than 0.0001). A total of 28 percent of acyclovir recipients and no placebo recipients remained free of recurrences for two years. The low rate of recurrences in the acyclovir group remained consistent throughout the study. No serious clinical or laboratory abnormalities associated with acyclovir were observed. The 800-mg acyclovir tablet given daily was well-tolerated and effective for two years in the management of recurrent genital herpes. PMID- 3044091 TI - Treatment of herpes labialis with acyclovir. Review of three clinical trials. AB - Three trials with acyclovir in the treatment of recurrent herpes labialis in the same patient population are reviewed. In the first trial, oral acyclovir capsules, five times per day for five days, were shown to be of significant benefit for some parameters of cutaneous resolution in three successive episodes. Topical formulations were evaluated in two separate trials; 5 percent acyclovir in a modified aqueous cream base exhibited favorable trends, but 5 percent acyclovir in a polyethylene base was ineffective. A high dose of oral acyclovir may offer the most effective method of control of recurrent herpes labialis. PMID- 3044092 TI - Preventive therapy of herpes labialis associated with trigeminal surgery. AB - Acyclovir was shown to limit herpes simplex reactivation in a controlled trial to prevent herpes labialis after surgical intervention for trigeminal neuralgia. Of 14 patients receiving acyclovir, unambiguous herpes labialis developed in only one, compared with 12 of 16 in the placebo group. PMID- 3044093 TI - Treatment of eczema herpeticum with oral acyclovir. AB - Clinical efficacy and safety of oral acyclovir for the treatment of eczema herpeticum were evaluated in 69 patients in a multicenter, double-blind placebo controlled trial. Patients were randomly assigned to receive either acyclovir, 200 mg five times per day for five days, or placebo. There were 32 evaluable patients in the acyclovir group and 28 in the placebo group. Clinical efficacy was assessed as very effective, effective, moderately effective, or not effective by using a numerical scoring for various parameters, such as lesion stage, pain, and general improvement. The efficacy rate was 81.3 percent in the acyclovir group and 42.9 percent in the placebo group (p less than 0.01), and significant differences were observed in general improvement and disappearance of ulcer. Results for duration of pain and erosion favored the acyclovir group, but did not reach statistical significance. Mild adverse reactions were documented in two patients receiving acyclovir and in six patients receiving placebo. Oral acyclovir seems to be a well-tolerated and effective therapy for eczema herpeticum. PMID- 3044094 TI - Management of mucocutaneous herpes simplex virus infections in immunocompromised patients. AB - Herpes simplex virus (HSV) infections are a significant cause of morbidity among immunocompromised patients, and in some instances these infections may be a primary cause of death. The overwhelming majority of these infections are caused by reactivation of the virus. The natural history of reactivated HSV infections in immunocompromised patients has been well described, and these infections occur predictably in particular patient populations. Antiviral therapy has been shown to be effective in treating or preventing HSV infections. Randomized, controlled, double-blind studies have demonstrated that acyclovir is the most effective compound currently available for treatment or prevention of HSV infections. PMID- 3044096 TI - Varicella zoster virus infections in immunocompromised hosts. A review of the natural history and management. AB - Varicella is relatively mild in otherwise normal children, in whom new lesions form for a mean of four days after onset and heal 50 percent of their lesions in eight days. New lesions form in most immunocompromised children for longer than five days and those not treated with antiviral drugs have a 28 percent incidence of pneumonitis and a 7 percent mortality rate. Untreated immunocompromised adults with herpes zoster shed virus for longer (7.0 days) than otherwise normal adults (5.3 days). Herpes zoster is much more likely to disseminate cutaneously in immunocompromised than in immunocompetent hosts. Visceral dissemination, which is a rare event in immunocompetent patients, occurred in 8 percent of prospectively followed untreated immunocompromised hosts with herpes zoster. Acyclovir has been found to be superior to vidarabine for treatment of both chickenpox and herpes zoster. Whether or not steroids should be used to treat herpes zoster remains controversial. Concerns about the use of intravenous acyclovir include the side effects of renal and central nervous system dysfunction and the possibility of emergence of resistant viral strains. None of these concerns has proved to be an impediment to successful treatment of immunocompromised patients. The major future challenge is to find an optimal way to treat varicella zoster virus infections with oral formulations of acyclovir or its congeners. PMID- 3044095 TI - Herpes simplex virus infections of the central nervous system. A review. AB - Herpes simplex virus (HSV) infections of the central nervous system are a significant cause of mortality and morbidity. The introduction of antiviral therapy has improved the outcome for patients with life-threatening disease. Neonatal HSV infection is usually acquired at the time of delivery by contact of the fetus with infected maternal genital secretions resulting in disease that can be localized to the skin, eye, and mouth, and can lead to encephalitis or become disseminated. A total of 291 babies with neonatal HSV infection have been evaluated over a period of 14 years with mortality and morbidity rates determined at one year. Vidarabine therapy decreased the incidence of mortality and improved morbidity rates; however, further improvement in mortality rates with acyclovir therapy has not been apparent. No significant clinical toxicity appeared in either treatment group. In order to improve outcome, earlier intervention and prophylactic strategies must be developed. For patients with herpes simplex encephalitis, acyclovir therapy is superior to vidarabine therapy for biopsy proven disease. When outcome is compared for 136 vidarabine- and 46 acyclovir treated, biopsy-proven patients, mortality rates are decreased to 20 percent with acyclovir, and approximately 40 percent of survivors are evaluated as normal at one year after therapy. Despite better outcome with antiviral therapy for the treatment of biopsy-proven herpes simplex encephalitis, further improvement is required. PMID- 3044097 TI - Management of varicella zoster infections in immunocompetent hosts. AB - Varicella in otherwise healthy children usually requires no antiviral treatment. In severe cases, however, such as are seen in neonates and adults, treatment must be given. Anecdotal evidence suggests the efficacy of intravenous acyclovir in such patients. Herpes zoster in immunocompetent patients may be severe enough to warrant antiviral therapy, particularly in elderly patients. Both idoxuridine and acyclovir have been investigated in placebo-controlled double-blind studies. Due to its low toxicity, ease of administration, and the possibility of systemic administration, acyclovir has largely replaced older antivirals in the management of herpes zoster in the normal host. Recent studies have shown the efficacy of oral acyclovir. In addition, oral acyclovir may prevent the ocular complications of ophthalmic zoster. When acyclovir is given, it should be administered as early as possible, preferably no later than four days after the onset of the rash. The combination of acyclovir and prednisolone for the prevention of post-herpetic neuralgia has not proved effective. PMID- 3044098 TI - Efficacy of oral acyclovir treatment of acute herpes zoster. AB - Oral acyclovir, 800 mg five times per day for seven days, was compared with placebo in a randomized, double-blind trial conducted at three centers in the United Kingdom. The study group consisted of 364 elderly immunocompetent patients with herpes zoster who were entered within 72 hours of the onset of rash. Acyclovir significantly reduced the times to last new lesion formation (p less than 0.01), loss of vesicles (p less than 0.01), and full crusting (p = 0.03). No significant hastening of rash healing was seen in those who started therapy later than 48 hours after the onset of rash. There was also a significant reduction pain during treatment with acyclovir (p = 0.02). Acyclovir produced no effects on the frequency or severity of post-herpetic neuralgia. No clinically important adverse effects of acyclovir were reported. PMID- 3044099 TI - Therapy of herpes zoster with oral acyclovir. AB - Oral acyclovir therapy for herpes zoster has been studied in double-blind, placebo-controlled trials of two dosages, 400 mg and 800 mg five times per day for 10 days. Compared with placebo recipients, recipients of the high-dosage acyclovir experienced a significantly shortened period of viral shedding, significantly accelerated time to 50 percent scabbing, significantly accelerated time to 50 percent healing, and after two days of therapy, significantly less frequent formation of new lesions. The duration and severity of acute pain were less in acyclovir recipients, with differences in pain severity achieving statistical significance (p = 0.03) between Days 3 and 10 and correlating with the treatment differences in new lesion formation. In studies of the 400 mg five times per day dose schedule, differences between acyclovir and placebo recipients were not significant. In a six-month follow-up of recipients in the higher dosage study, the acyclovir recipients experienced less post-zoster pain than placebo recipients; differences in the prevalence of pain were most significant for the presence of a persistent pain in the first three months of follow-up. Oral acyclovir at these dosages appears to be free of adverse reactions. In summary, oral acyclovir at a dosage of 800 mg five times per day for 10 days for treatment of acute herpes zoster is superior to 400 mg five times per day and favorably alters the course of the disease. PMID- 3044100 TI - Reduction of the ocular complications of herpes zoster ophthalmicus by oral acyclovir. AB - Herpes zoster ophthalmicus (HZO) is a unique form of zoster dermatitis associated with a high rate of ocular complications that tend to be chronic and may cause vision loss. The ocular complications are highly varied, with keratitis and uveitis being more persistent sequellae of HZO. Oral acyclovir treatment of acute HZO reduces the incidence of the more common ocular complications, including keratitis and uveitis. Although patients treated early in the course of this disease experience a greater clinical response, treatment as late as seven days after onset of cutaneous lesions confers a beneficial prophylactic effect with respect to the ocular complications of HZO. PMID- 3044101 TI - Treatment of chickenpox in immunocompromised children. AB - In Hungary since 1981, 98 immunocompromised children have been treated with intravenous acyclovir for varicella zoster virus infections. They were treated in an open study or a double-blind study. Results of both are discussed briefly. Overall, five of 74 patients with varicella died, whereas all 24 patients with herpes zoster recovered. Treatment failures occurred in patients in whom treatment started late and in those with severe lymphocytopenia. In some children, varicella zoster virus-specific antibodies failed to develop by the end of treatment, and a proportion of these suffered recurrent episodes of varicella. PMID- 3044102 TI - Current therapy of varicella zoster virus infection in immunocompromised patients. A comparison of acyclovir and vidarabine. AB - Both acyclovir and vidarabine are effective treatment for varicella zoster virus (VZV) infection in immunosuppressed patients. To determine which is preferable, therapy with these two agents was compared in a prospective, randomized trial. A total of 22 immunocompromised patients undergoing treatment for hematologic malignancies and presenting with VZV infection within 72 hours of the onset of rash were randomly assigned to receive intravenous acyclovir or vidarabine; 11 patients were randomly assigned to each treatment group. Acyclovir was significantly more effective than vidarabine in preventing complications of VZV infection, and treatment failures requiring a change to the alternate therapy occurred only among those treated with vidarabine. As compared with vidarabine, acyclovir shortened the median period during which results of viral culture specimens were positive and new lesions formed. Acyclovir also shortened the median interval until the first decrease in pain, the crusting of all lesions, and the complete healing of lesions. Acyclovir is more effective than vidarabine in the treatment of VZV infection in severely immunocompromised patients and should be considered the treatment of choice in such cases. PMID- 3044103 TI - Prevention of herpes zoster in patients by long-term oral acyclovir after allogeneic bone marrow transplantation. AB - Following allogeneic bone marrow transplantation for leukemia, herpes zoster infections that are potentially severe with a high risk of dissemination develop in 30 to 50 percent of patients. Intravenous acyclovir is an effective treatment for established zoster in immunocompromised persons. Oral acyclovir has relatively low bioavailability, which has made the value of this route of administration for the treatment or prophylaxis of herpes zoster infections uncertain. In this trial, 82 patients undergoing allogeneic bone marrow transplantation for leukemia were randomly assigned to receive either intravenous acyclovir for 23 days followed by oral acyclovir for six months, or matched placebos; the random groups were well-matched in all clinical characteristics. During the six-month period of acyclovir/placebo administration, no patient receiving acyclovir developed herpes zoster, whereas six patients receiving placebo did so (p = 0.006). During the six-month follow-up, there were six cases of zoster in the treatment arm of the study and two cases in the placebo arm. Herpes zoster was not restricted to those patients who had positive evidence of antibody before transplant. This study shows that oral acyclovir is capable of preventing zoster infection during its period of administration; once the drug treatment is stopped, infections occur. In selected patients, the use of long term oral acyclovir may be of value in preventing zoster infections during the time of greatest immunosuppression. PMID- 3044104 TI - Respiratory complications of cardiac transplantation. AB - The authors evaluated all respiratory complications of cardiac transplantation in a 10-year study of 94 consecutive recipients. Mean follow-up time was 20 +/- 17 months. The initial 20 patients were treated with azathioprine and prednisone, while the subsequent 74 patients received cyclosporine and prednisone. In the azathioprine group, respiratory infections accounted for 24 of 60 (40%) infections. Two-thirds of the respiratory infections occurred in the first 3 postoperative months and were generally localized processes (focal pneumonitis, nodule(s), abscess, or empyema). Gram-positive and gram-negative bacteria (8/30) and aspergillus (8/30) were the predominant pathogens. Respiratory failure occurred in 29% of infectious episodes. In the cyclosporine group, there were significantly fewer respiratory infections. There was also a reduction in the number of nonrespiratory infections; hence, the percentage of total infections due to respiratory causes, 26 of 50 (52%), was not significantly different. In contrast, however, nearly two-thirds of the respiratory infections in cyclosporine-treated patients occurred after the first 3 postoperative months, and were usually diffuse processes. Despite diffuse disease, respiratory failure was observed with similar frequency (19%). Pneumocystis carinii (9/31) and cytomegalovirus (CMV) (7/31) were the predominant pathogens. CMV pneumonitis tended to occur earlier than that due to P. carinii (2.9 +/- 1.9 mo vs. 9.8 +/- 11.2 mo, respectively), but there was considerable overlap. In comparison with infectious processes, there were 50% fewer noninfectious respiratory complications in both groups. These were primarily pleural (46%) or thromboembolic (18%) disorders. Four of five pulmonary emboli occurred in patients with intercurrent cardiorespiratory illness, and were detected only at autopsy. The authors conclude that respiratory infections account for one-half of all infections observed in cardiac transplant recipients, despite the reduced infection rate associated with the use of cyclosporine. Furthermore, respiratory infections in cyclosporine-treated patients exhibit different clinical and etiologic features than those seen in azathioprine-treated patients. Finally, occult thromboemboli may be difficult to recognize in cardiac transplant recipients because of the high incidence of coexisting cardiorespiratory disease. PMID- 3044105 TI - Oral terbutaline augments cardiac performance in chronic obstructive pulmonary disease. AB - In previous research, we have demonstrated that parenterally administered terbutaline can augment resting cardiac function in patients with chronic obstructive pulmonary disease (COPD). Because the oral form of terbutaline is more widely utilized, a double-blind, randomized, crossover, placebo-controlled trial of the cardiopulmonary effects of oral terbutaline was conducted in ten patients with COPD. Right and left ventricular ejection fractions (RVEF and LVEF) were determined by first pass radionuclide angiography. There were no differences in spirometry and hemodynamic measurements between treatment and placebo days. Following 5 mg of oral terbutaline, there was a small but statistically significant increase in forced expiratory volume in 1 second and in heart rate, but no significant change in forced vital capacity or blood pressure. LVEF improved significantly with terbutaline both at rest (62% +/- 6% vs. 67% +/- 9%, mean +/- SD) and during submaximal steady state exercise (61% +/- 5% vs. 67% +/- 10%). RVEF improved significantly at rest (64% +/- 6% vs. 69% +/- 5%), but not during submaximal steady state exercise (65% +/- 6% vs. 68% +/- 7%). Thus, oral terbutaline produces significant improvement in biventricular systolic pump performance at rest, and increases left ventricular ejection fraction during submaximal exercise in patients with moderate to severe COPD. PMID- 3044106 TI - Cholesterol gallstone disease: the current status of nonsurgical therapy. AB - Gallstone disease is a common disease that appears to be related to a Western diet. The underlying pathogenesis is a subtle alteration in the liver such that excessive cholesterol is extracted from the liver cell by bile acids undergoing an enterohepatic recirculation. Gallstone disease progresses through well-defined stages, beginning with a bile supersaturated with cholesterol and proceeding to crystal formation, stone growth, and finally symptoms caused by impaction of a stone in either the cystic duct or the common bile duct. The natural history is that most stones never cause symptoms. Stones that cause symptoms have been present for an average of 12 years. The treatment of truly asymptomatic stones should be observation. Ultrasonography of the right upper quadrant is the gold standard for the diagnosis of stones in the gallbladder. Endoscopic retrograde cholangiopancreatography (ERCP) is the gold standard for the diagnosis of stones in the common bile duct. Oral cholecystogram (OCG) helps select patients who have noncalcified, floating stones that may be dissolved with bile acids or methyl tertiary butyl ether (MTBE). Therapy with chenodiol has been a disappointment because of a low complete response rate. The ideal candidate for attempted dissolution with chenodiol would be a thin woman with hypercholesterolemia and a small number of symptomatic, small, floating, radiolucent gallstones. Ursodeoxycholic acid (Urso), when it is available, will have all of the attributes of chenodiol and virtually none of the side effects. Rapid dissolution of gallstones with MTBE shows great promise of being a generally available means of dissolving gallstones. Extracorporeal shock wave lithotripsy also shows promise, but its general availability may be limited by the cost of the equipment needed. As of now, the treatment of choice for symptomatic gallstones remains cholecystectomy, unless there is a compelling reason not to operate. PMID- 3044107 TI - Steroid receptors, cellular kinetics, and lymph node status as prognostic factors in breast cancer. AB - Steroid receptor status, cellular kinetics, abnormal proto-oncogene presence, and lymph node metastases all have been shown to provide prognostic information in breast cancer. The factors guide the choice of therapy and predict the course of the disease. Both disease-free survival and overall survival are predicted by these variables. Steroid receptors are the most reliable predictor of hormonal responsiveness. Lymph node involvement is crucial in determining the extent of the disease and the need for adjuvant therapy. Cellular kinetics and abnormal proto-oncogene presence predict tumor aggressiveness. Together these prognostic factors provide considerable information to the clinician. PMID- 3044108 TI - Clinical significance of elevated mean arterial pressure in the second trimester. AB - Several reports suggested using the mean arterial blood pressure during the second trimester to predict the future development of preeclampsia. The value of a second-trimester mean arterial blood pressure greater than or equal to 90 mm Hg was reviewed in 39,876 reported cases of preeclampsia and 207 cases of eclampsia. The sensitivity ranged from 0% to 92% and the specificity varied from 53% to 97%. The predictive value of a positive test ranged from 0% to 43% and the predictive value of a negative test ranged from 76% to 98%. The predictive value of a positive test was not greatly higher than the incidence of hypertension in the whole population studied in the majority of the reports. There was a strong association of higher second-trimester mean arterial blood pressure in nulliparous women with eclampsia who had transient hypertension in later gestations and ultimately developed chronic hypertension. The data suggest that one sign of future chronic hypertension, that is, transient hypertension, is often predicted by high second-trimester mean arterial blood pressure, which may have the same significance. We think that if increased second-trimester mean arterial blood pressure levels predict anything, it is transient hypertension rather than preeclampsia-eclampsia. PMID- 3044109 TI - The sperm penetration assay: can it discriminate between fertile and infertile men? AB - The sperm penetration assay with zona-free hamster ova is widely used to evaluate male infertility. Despite a growing body of literature about this assay, its results remain difficult to interpret. To evaluate the clinical usefulness of this test, we reviewed the world's literature about this assay and analyzed the test's performance. Its sensitivity ranges from 0.00 to 1.00 and specificity ranges from 0.95 to 1.00 for diagnosing male infertility. For predicting in vitro fertilization failures, its sensitivity varies from 0.00 to 0.78 and specificity ranges from 0.51 to 1.00. The reproducibility of this assay is not clear, and there is little standardization of methods between laboratories. Until the validity and reproducibility of the sperm penetration assay have been established, this expensive test should probably not be used to evaluate infertile couples. PMID- 3044110 TI - Sodium excretion in normal and hypertensive pregnancy: a prospective study. AB - One hundred fifty-eight intravenous saline solution infusions (3 mmol Na per kilogram body weight) were performed in (1) normal primigravid women during the second and third trimesters and post partum, after 1 week of either a high, low, or ad libitum salt intake; (2) normotensive primigravid women during midpregnancy who later developed pregnancy-induced hypertension, and (3) seven proteinuric and seven nonproteinuric primigravid women with ad libitum salt intake who had established pregnancy-induced hypertension. Sodium excretion did not differ significantly between pregnancy and after pregnancy despite marked differences in plasma renin activity, aldosterone concentration, volume, and glomerular filtration rate. Sodium excretion after saline solution loading varied according to prestudy sodium intake and was reduced between the second and third trimesters, independent of dietary salt intake. Those destined to develop pregnancy-induced hypertension had sodium excretion similar to that of continuously normotensive subjects during the second trimester, but those with established proteinuric pregnancy-induced hypertension had the lowest plasma volume, plasma aldosterone concentration, and plasma renin activity and retained sodium to the same degree as salt-deplete women with normotension. These results demonstrate that the balance of sodium regulatory factors is similar between pregnancy and post partum, that prestudy salt intake and stage of gestation can alter the natriuretic response to saline solution loading, and that normal pregnant women retain more administered sodium in late pregnancy than in midpregnancy despite further increases in plasma volume and no alterations to blood pressure or glomerular filtration rate. Those with established proteinuric pregnancy-induced hypertension retain sodium avidly without stimulation of plasma renin activity or plasma aldosterone concentration, findings not apparent during midpregnancy in those who later developed this disorder. PMID- 3044111 TI - Intramuscular versus intravenous ritodrine hydrochloride for preterm labor management. AB - A prospective, interinstitutional comparative trial was undertaken to examine the efficacy, safety, and pharmacodynamics of different administration routes of ritodrine hydrochloride for the management of preterm labor. Forty-five subjects between 20 and 36 weeks' gestation received either intravenous (n = 24) or intramuscular (n = 21) therapy. Successful tocolysis occurred in 14 of 21 (67%) patients in the group treated intramuscularly and in 16 of 24 (67%) patients in the group treated intravenously. A greater mean dose (8.6 versus 3.3 mg/hour) and a higher mean serum concentration (38.9 versus 24.7 micrograms/ml) were needed to achieve successful tocolysis in the intravenous group as compared with the intramuscular group. Patients who did not respond to tocolytic therapy in both groups had levels of ritodrine in the blood either equivalent to or greater than those of subjects who were successfully treated. Analysis of ritodrine levels in the successfully treated intramuscular group demonstrated significant differences in blood levels depending on muscle group used. These differences can be at least partially attributed to higher mean doses administered to patients receiving vastus lateralis injections as compared with those receiving gluteal muscle injections. The results suggest that intramuscular administration of ritodrine is an efficacious and safe route of drug delivery. Additional studies are needed to better define dose-response curves for the intramuscular administration of ritodrine hydrochloride. PMID- 3044112 TI - Early diagnosis of pregnancy failure with transvaginal ultrasound. AB - Vaginal ultrasound was used to assess a consecutive series of 353 patients to determine the minimum mean gestation sac diameter at which a failed pregnancy could be diagnosed. Fetal heart movements could be demonstrated in all pregnancies with living fetuses when the mean sac diameter exceeded 1.2 cm. Vaginal ultrasound is superior to both static and transabdominal real-time ultrasound in the diagnosis of early pregnancy failure. PMID- 3044113 TI - De novo clinical hypothyroidism in pregnancies complicated by type I diabetes, subclinical hypothyroidism, and proteinuria: a new syndrome. AB - Fifty-one women with type I diabetes who had normal thyroxine values before becoming pregnant were evaluated. Abnormalities of thyroid tests other than thyroxine were encountered in 26 women, of whom 8 developed a low serum thyroxine level, an elevated thyroid-stimulating hormone level, and a low insulin requirement in the second trimester subsequent to an increase in 24-hour urinary protein excretion to greater than 4 gm/24 hr. Thyroid replacement led to an increase in insulin requirement to levels appropriate for gestational age. It is concluded that the woman with type I diabetes who develops proteinuria greater than 4 gm/24 hr during gestation is at risk for the development of de novo hypothyroidism during pregnancy, evidenced by a low serum thyroxine level, an elevated thyroid-stimulating hormone level, and a drop in insulin requirement. PMID- 3044114 TI - An association between hyperinsulinemia and hypertension during the third trimester of pregnancy. AB - We studied 43 women in their third trimester of pregnancy whose fetuses were at significant risk of intrauterine growth retardation. To define metabolic subtypes for intrauterine growth retardation, a 100 gm glucose load was administered after an overnight fast. Twenty-seven women were normotensive and 16 had hypertension. The glucose tolerance of the hypertensive group was essentially the same as that of the normotensive group. However, 8 of the 16 women with hypertension had marked hyperinsulinemia in response to an oral glucose load. Of the five women with hypertension who gave birth to offspring of low birth weight, three had hyperinsulinemia. PMID- 3044115 TI - Fetal lung maturity in diabetic pregnancies: relation among amniotic fluid insulin, prolactin, and lecithin. AB - Insulin, prolactin, and lecithin phosphorus levels were measured in 97, 62, and 44 amniotic fluid samples from third trimester normal, gestational diabetic, and insulin-dependent diabetic patients, respectively. There was no difference in lecithin phosphorus concentration (index of fetal lung maturity) among the three groups. The amniotic fluid insulin level was significantly higher in insulin dependent diabetic patients, whereas there was no difference in amniotic fluid prolactin levels among the groups. Correlations of amniotic fluid prolactin levels with both lecithin phosphorus and insulin levels were not statistically significant in any of the groups. This is probably because amniotic fluid prolactin is decidual, rather than fetal, in origin. Even though amniotic fluid insulin levels, which reflect fetal levels, were significantly higher in insulin dependent diabetic patients, there was no difference in the amniotic fluid lecithin phosphorus concentration in diabetic pregnancies compared with that in normal pregnancies. Moreover, there was a positive, and not a negative, correlation between amniotic fluid insulin and amniotic fluid lecithin phosphorus levels in diabetic pregnancies. These results do not support the theory that fetal hyperinsulinemia results in delayed pulmonic maturation in diabetic pregnancies. PMID- 3044116 TI - Human leukocyte antigen compatibility in a couple with idiopathic recurrent hydramnios. AB - Recurrent hydramnios of unknown cause is a very rare obstetric complication. Only three cases have been reported. We report a case of idiopathic recurrent hydramnios associated with human leukocyte antigen compatibility between the patient and her husband. PMID- 3044117 TI - The prenatal sonographic diagnosis of lethal multiple pterygium syndrome: a heritable cause of recurrent abortion. AB - Presumably, a large number of recurrent abortions are caused by lethal recessive syndromes whose diagnosis depends either on a known family history or on the identification of characteristic fetal phenotypic features on pathologic examination. Because these conditions are rare, family histories are seldom helpful, and nondirected postmortem examinations on degenerating samples are seldom enlightening. Serial ultrasonography beginning early in pregnancy may provide important information in the evaluation of recurrent abortion caused by lethal recessive disorders. Reported is the accurate prenatal sonographic diagnosis of lethal multiple pterygium syndrome in a patient with a history of recurrent abortions. This syndrome is characterized by multiple limb contractures with pterygia, facial clefting, intracranial abnormalities, cystic hygroma, progressive fetal edema, and fetal death by midgestation. Inheritance may be X linked recessive. Lethal multiple pterygium syndrome should be considered in patients with a history of recurrent midtrimester losses. PMID- 3044118 TI - Management of heart transplant recipients: guidelines for the obstetrician gynecologist. AB - As the number and survival time of heart transplant recipients continue to increase, their quality of life, including sexuality and childbearing, have become important issues. Reproduction is possible for both male and female patients after the transplant. Counseling for contraception when sterilization is not desired must take into account the increased risk of infection and genital carcinoma associated with immunosuppressant drug therapy. Teratogenicity has not been reported either with traditional immunosuppressive agents (prednisone, azathioprine) or with cyclosporine. Osteoporosis prophylaxis is particularly important in the female heart transplant recipient, because the chronic use of prednisone increases this risk. Guidelines are provided to counsel patients in these areas. PMID- 3044120 TI - Immunocytochemical localization of angiotensin II immunoreactivity and demonstration of angiotensin II binding in the rat ovary. AB - The cellular localization of angiotensin II immunoreactivity and the presence of angiotensin II binding in rat ovaries were studied. Angiotensin II immunohistochemical staining was demonstrated throughout the corpora lutea of gonadotropin-stimulated immature rats and pseudopregnant adult rats, as well as in some stromal and thecal cells surrounding large antral follicles. No immunostaining was observed in granulosa cells of preantral or antral rat follicles or in ovaries from unstimulated immature rats. With in vitro autoradiography, specific, saralasin-suppressible 125I-angiotensin II binding was demonstrated in normal cycling rat ovaries: diestrus greater than proestrus greater than estrus. The combined findings of angiotensin II immunostaining in ovarian follicles and corpora lutea and of cycle-related angiotensin II binding support the hypothesis of a functional role for the ovarian renin-angiotensin system. PMID- 3044119 TI - Vitamin B12 R-binder localization in the human uterus: an immunohistochemical study. AB - The localization of vitamin B12 R-binder in the uterus was studied by use of an immunoperoxidase technique. Positive staining by anti-R-binder antiserum was observed in the columnar epithelium of the endocervix (18/18 cases) and in the surface epithelium of the endometrium (8/21 cases). Staining was usually seen in the apical portion of the epithelium; cytoplasmic staining in the endocervical columnar epithelium was intense. The secretory products in the endocervical glands showed positive staining. The endometrial glandular epithelium did not stain (0/24 cases). Metaplastic squamous epithelium of the endocervix showed positive staining (3/18 cases). The native squamous epithelium as well as the stromal components of the cervix, endometrium, and myometrium were negative for R binder. This study shows that R-binder is localized in the uterus, especially in the endocervical glands. The R-binder in the endocervix may have antimicrobial activity in the uterus as in other organs, such as the intestines and mammary glands. PMID- 3044121 TI - Ultrasound determination of nuchal cord in breech presentation. PMID- 3044122 TI - Is hygiene enough? Circumcision as a possible strategy to prevent neonatal group B streptococcal disease. PMID- 3044124 TI - Meredith Walter Morgan--a salute. PMID- 3044123 TI - Festschrift in honor of Meredith W. Morgan. PMID- 3044125 TI - Effect of vision therapy on stereophotogrammetric profiling--a controlled clinical trial. AB - A single, masked controlled clinical trial showed that vision therapy improved both the accuracy and repeatability of stereophotogrammetric performance on the profiling task with experienced, visually normal observers. The average fixation disparity measured under working conditions decreased and stereoscopic acuity as measured with a Howard-Dolman apparatus increased. The data suggest that vision therapy is most helpful for those profiling situations in which disparity stimuli are plentiful and stereopsis is the dominant depth cue. PMID- 3044127 TI - Presbyopia in light of accommodation. AB - The "dual, indirect, active" mechanisms of accommodation proposed by Helmholtz is reviewed. New supporting evidence shows that the insertion of the ciliary muscle is onto the span fibrils of the zonule in the interciliary process regions of the ciliary body. Thus, the peripheral zonule is the elastic antagonist of a unified ciliary muscle. A biomechanical model illustrating accommodation is put forward that makes consistent and explicit the force and length changes that the six mechanical component elements undergo. The Hess-Gullstrand lenticular theory of presbyopia is compatible with this model. PMID- 3044126 TI - Optometry of Daza de Valdes (1591-ca. 1636) AB - The 1623 Spanish book entitled Uso de los Antoios by Benito Daza de Valdes has been translated into English and is presently available in manuscript form at the International Library, Archives, and Museum of Optometry. The first of its three parts deals with the nature and properties of the eye and its refractive conditions. The second discusses contemporary spectacle lenses and frames in technological and utilitarian detail. The third and major part in the style of Platonian dialogue describes a variety of clinical optometric problems and their practical solutions. The procedural similarities to today's mode of optometric analysis is remarkably evident. PMID- 3044128 TI - Pupil dependency of bifocal contact lenses. AB - The advantages and the problems associated with use of alternating vision and simultaneous vision bifocal contact lenses are analyzed. The changes in pupil diameter associated with changes in illumination and in the distance of the task make the selection of the best bifocal design complex. PMID- 3044129 TI - The medical and social science basis for a national infant care leave policy. AB - In a review of work in psychology, psychiatry, neonatology, and sociology, the birth or adoption of an infant emerges as a sensitive period in which parents need time for role adaptation. Evidence points to the need for job-protected leave without financial loss so that parents can meet the material and emotional needs of their family in general, and the developmental needs of their new infant in particular, during the early postpartum period. PMID- 3044130 TI - Race, class, and adolescent pregnancy: an ecological analysis. AB - The U.S. has one of the highest rates of adolescent pregnancy and childbearing in the industrialized world--and the highest rates in the U.S. are found among low income black adolescents. This paper addresses the problem via a four-part theoretical framework based on an ecological developmental model. Variables that contribute to adolescent pregnancy in the black community are examined at the individual, family, sociocultural, and social structural levels. The potential utility of this framework is discussed, and suggestions are offered for research and programmatic intervention. PMID- 3044131 TI - Sex differences in children's response to parental divorce: 1. Research methodology and postdivorce family forms. AB - This two-part review documents inconsistencies in the empirical basis for the hypothesis that boys are more negatively affected than are girls by parental divorce. An attempt is made to move beyond a global hypothesis to one specifying circumstances for the pattern. Part 1 asks whether methodological adequacy or postdivorce family forms in the studies help explain the discrepancy across studies that examine sex differences in children's response to parental divorce. Part 2, reviewing other possible sources for the discrepancy, will appear in a forthcoming issue of this Journal. PMID- 3044133 TI - Cochlear implants in children: nonmedical considerations. PMID- 3044132 TI - Child custody mediation: an interdisciplinary synthesis. AB - The high divorce rate and increasing number of children affected by divorce have paralleled major changes both in divorce law and in the ways in which social conflicts are resolved. Principles of contemporary law and conflict resolution are joined with the findings of research on child development in families of divorce in a conceptual synthesis relevant to the practice of child custody mediation. PMID- 3044134 TI - Effects of English admixture and geographic distance on anthropometric variation and genetic structure in 19th-century Ireland. AB - The analysis of anthropometric data often allows investigation of patterns of genetic structure in historical populations. This paper focuses on interpopulational anthropometric variation in seven populations in Ireland using data collected in the 1890s. The seven populations were located within a 120-km range along the west coast of Ireland and include islands and mainland isolates. Two of the populations (the Aran Islands and Inishbofin) have a known history of English admixture in earlier centuries. Ten anthropometric measures (head length, breadth, and height; nose length and breadth; bizygomatic and bigonial breadth; stature; hand length; and forearm length) on 259 adult Irish males were analyzed following age adjustment. Discriminant and canonical variates analysis were used to determine the degree and pattern of among-group variation. Mahalanobis' distance measure, D2, was computed between each pair of populations and compared to distance measures based on geographic distance and English admixture (a binary measure indicating whether either of a pair of populations had historical indications of admixture). In addition, surname frequencies were used to construct distance measures based on random isonymy. Correlations were computed between distance measures, and their probabilities were derived using the Mantel matrix permutation method. English admixture has the greatest effect on anthropometric variation among these populations, followed by geographic distance. The correlation between anthropometric distance and geographic distance is not significant (r = -0.081, P = .590), but the correlation of admixture and anthropometric distance is significant (r = 0.829, P = .047). When the two admixed populations are removed from the analysis the correlation between geographic and anthropometric distance becomes significant (r = 0.718, P = .025). Isonymy distance shows a significant correlation with geographic distance (r = 0.425, P = .046) but not with admixture distance (r = -0.052, P = .524). The fact that anthropometrics show past patterns of gene flow and surnames do not reflects the greater impact of stochastic processes on surnames, along with the continued extinction of surnames. This study shows that 1) anthropometrics can be extremely useful in assessing population structure and history, 2) differential gene flow into populations can have a major impact on local genetic structure, and 3) microevolutionary processes can have different effects on biological characters and surnames. PMID- 3044135 TI - Hyperostosis frontalis interna: a Nubian case. AB - The aim of this article is to present evidence of hyperostosis frontalis interna in a 40-year-old female recovered from a Meroitic cemetery (ca. 300 A.D.) in Sudanese Nubia. A review of the literature concerning the Morgagni-Stewart-Morel (MSM) syndrome suggests that the changes in the skull fragment are consistent with this diagnosis. This case is the earliest example of the condition so far reported, and therefore, in archaeological time and space, this is a disease not only of modern civilization, but also of antiquity. Current endocrinological reports suggest that the hyperostosis is the hallmark of a generalized disorder of bone metabolism, with increased androgens, prolactin, and somatotropins. Hyperostosis frontalis interna is the central feature of a syndrome first described over 200 years ago by the early pathologist Giovanni Batistta Morgagni, professor of anatomy at Padua (1719). He found thickening of the internal tables of the frontal bones in association with virilism and obesity. Stewart (1928) and Morel (1929) independently added several neuropsychiatric problems to this complex and questioned the possibility of an endocrine basis for the syndrome. PMID- 3044137 TI - Effect of thyroid hormones on oxidative and nonoxidative glucose metabolism in humans. AB - The glucoregulatory function of thyroid hormones was investigated in six healthy subjects before and after 14 day 3,5,3',5'-tetraiodothyronine (T4) treatment (300 micrograms/day) using a sequential clamp protocol for 5 h at euglycemia (0-2 h) and hyperglycemia (165 mg/dl, 2-5 h) and different insulin infusion rates (1.0 for 0-3.5 h and 6.5 mU.kg-1.min-1, for 3.5-5 h). T4 treatment increased basal energy expenditure (+8%), glucose disposal (+31%), and oxidation (+87%) but decreased nonoxidative glucose metabolism (-30%) and was without effect on lipid oxidation. During the euglycemic clamp, T4 treatment enhanced insulin-induced glucose disposal (+16%), glucose oxidation (+34%), and inhibition of lipid oxidation (-66 vs. -40%); nonoxidative glucose metabolism was stimulated to a similar extent before and after T4. During hyperglycemia, 3,5,3'-triiodothyronine (T3) did not affect glucose disposal but increased carbohydrate-induced lipogenesis at both insulin infusion rates. We conclude that T4 treatment promotes glucose disposal and oxidation, T3 decreases noninsulin-mediated glucose storage but does not antagonize insulin action. PMID- 3044136 TI - Effect of hormones on growth and function of cultured canine tracheal epithelial cells. AB - Insulin (INS), endothelial cell growth supplement (ECGS), transferrin (TF), cholera toxin (CT), hydrocortisone (HC), triiodothyronine (T3), and epidermal growth factor (EGF) were systematically examined for their effects on proliferation, ion transport activity, and morphological differentiation of canine tracheal epithelial (CTE) cells in culture. INS, ECGS, TF, and CT increase proliferation of CTE cells cultured on plastic Petri dishes but exhibit no acute effect on the bioelectric activity of freshly excised CTE. CT increases amiloride insensitive transepithelial ion transport across both freshly excised canine trachea and CTE cultures. EGF has no effect on proliferation of CTE cells on plastic but induces hyperplasia of CTE cells on collagen matrices. EGF does not alter basal transepithelial ion transport across cultures but increases cellular responsiveness to beta-adrenergic stimulation. HC and T3 have no effect on proliferation or transepithelial bioelectric properties of CTE cells but improve morphological differentiation yielding cultures of complex epithelia containing cuboidal ciliated and nonciliated cells. These results demonstrate that growth and function of cultured CTE cells are affected by specific growth factors. PMID- 3044138 TI - Pancreatic islet discrimination of hexose anomers. II. Transient computer simulation. AB - We have previously modeled pancreatic islet glycolysis under idealized steady state conditions where the input is a pure hexose anomer and there is no mutarotation and reproduced the known preference for the alpha-anomers of glucose and mannose as substrates. This model is here extended to simulate real experiments, where the hexoses mutarotate and measurements may be taken over time. The behavior of our model system agrees with available experimental data. The hexose diphosphate activators of phosphofructokinase, whose effect was seen as not important in the preceding steady-state analysis, are found here to have a modest (approximately 10-15%) effect on its flux. The previous conclusion that the anomeric preference of the glycolytic pathway follows from that of glucokinase continues to hold in the real experimental situation. PMID- 3044139 TI - Hormonal and metabolic response to recombinant human tumor necrosis factor in rat: in vitro and in vivo. AB - Tumor necrosis factor (TNF; cachectin) has been implicated as a mediator of the toxic manifestations of overwhelming bacterial infection as well as the chronic catabolic state of cancer cachexia. We have examined the acute metabolic and hormonal response after administration of recombinant human TNF in the rat. TNF given by intraperitoneal injection produced dose- and time-related increases in hepatic amino acid uptake, decreases in serum trace metal concentrations, and a pattern of endocrine hormone alterations characteristic of the acute phase response to tissue injury. In vitro zinc transport studies by rat hepatocytes cultured in the presence of TNF alone, or in combination with recombinant human interleukin 1, another mediator of the acute phase response, demonstrated that neither monokine was capable of directly stimulating zinc transport into cells. These findings suggest that TNF may function as an endogenous mediator of the early metabolic response to sepsis and that the trace metal changes induced by TNF in vivo may occur through a secondary mechanism. PMID- 3044140 TI - Regulatory signals for intestinal amino acid transporters and peptidases. AB - Dietary protein ultimately regulates many processes involved in protein digestion, but it is often unclear whether proteins themselves, peptides, or amino acids (AAs) are the proximate regulatory signal. Hence we compared several processes involved in protein digestion in mice adapted to one of three rations, identical except for containing 54% of either casein, a partial hydrolysate of casein, or a free AA mixture simulating a complete hydrolysate of casein. We measured brush-border uptakes of seven AAs that variously serve as substrates for four AA transporters, and brush-border and cytosolic activities of four peptidases. The three rations yielded essentially the same AA uptake rates. Peptidase activities tended to be lower on the AA ration than on the protein ration. In other studies, all three rations yielded the same rates of brush border peptide uptake; protein is only modestly more effective than AAs at inducing synthesis of pancreatic proteases; and, depending on the animal species, protein is either much less or much more effective than AAs at stimulating release of cholecystokinin and hence of pancreatic enzymes. Thus the regulators of each process involved in protein digestion are not necessarily that process's substrate. We call attention to other cases in which the functional significance of regulatory signals remains to be understood. PMID- 3044142 TI - Hyperresponsiveness of arterial rings induced by renal prehypertensive rabbit plasma. AB - Vascular rings from normal rabbit renal arteries, when bathed in processed plasma from 3-day renal artery stenosis (RAS) rabbits, had greater contractile responses to norepinephrine than did matched rings bathed in processed plasma from sham operated rabbits. This vascular hyperresponsiveness of the rings produced by 3 day RAS plasma was abolished by the angiotensin II (ANG II) antagonist [Sar1, Ile8] ANG II but not by [Sar1, Ala8] ANG II at a dose that completely blocked the contractile responses of the rings to ANG II. The addition of [Sar1, Ala8]-ANG II to rings bathed in normal rabbit plasma did not alter the contractile responses to norepinephrine, indicating a lack of agonistic action by this ANG II analogue. These studies demonstrated that the ANG II receptors involved in the hormonally mediated vascular hyperresponsiveness in 3-day RAS rabbits are contained in the tissues that comprise these renal arterial rings. PMID- 3044141 TI - Protein digestion in human and rat small intestine: role of new neutral endopeptidases. AB - Two new phosphoramidon-insensitive neutral endopeptidases were identified and partially characterized in the brush-border membrane of rat and human intestine using N-CBZ-L-Ala-L-Arg-L-Arg-4-methoxy-beta-naphthylamide (Z-Ala-Arg-Arg-MNA) and azocasein or alpha-casein as substrates. Activities in the brush-border membrane of both rat and human intestine were maximum at neutral to alkaline pH, were inhibited by metal chelating and thiol reagents, and were insensitive to phosphoramidon. The results also indicate that these endopeptidases are distinct from pancreatic proteases. The biochemical properties of the enzyme hydrolyzing Z Ala-Arg-Arg-MNA were shown to be different from that hydrolyzing azocasein or alpha-casein. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration revealed that several native intact protein substrates were rapidly degraded to small molecular weight peptides and amino acids when incubated with rat or human brush-border membrane preparations. During in vivo intestinal perfusion in rats, 11% of the total administered alpha-casein was hydrolyzed and absorbed by the intestine. The results suggest that phosphoramidon-insensitive endopeptidases in the intestinal brush-border membrane may be of nutritional and physiological importance in protein digestion. PMID- 3044144 TI - Angiotensin-related sodium appetite and thirst in cattle. AB - Cows depleted of Na by loss of saliva from a parotid fistula for 46 h had an avid appetite for Na solution. They drank 21.0 +/- 1.6 liter of 0.3 M NaHCO3-NaCl solution during 2 h of access but little or no water during that time. Solutions of angiotensin II or captopril were infused for 3 h intravenously or into a lateral ventricle (intracerebroventricular) beginning 1 h before access to Na solution. Intravenous angiotensin II increased Na intake (to 26.8 +/- 2.9 liter, P less than 0.01) but did not alter water intake. Intracerebroventricular angiotensin II increased water intake but did not alter Na intake. Intravenous captopril reduced Na intake (to 11.0 +/- 2.1 liter, P less than 0.001) and concurrent intravenous angiotensin II prevented the reduction but concurrent intracerebroventricular angiotensin II did not. Intracerebroventricular captopril did not alter Na or water intake. Intravenous captopril reduced to zero the water intake during the hour before Na access, and concurrent intravenous angiotensin II prevented that reduction also. The dipsogenic action of intracerebroventricular angiotensin II was potentiated by intravenous captopril. The results of these experiments suggest that if angiotensin II receptors involved in the mechanism regulating Na appetite are in the brain, they are accessible only from the blood, e.g., in circumventricular organs. Thirst was inhibited by reduction of angiotensin II in blood but was stimulated only by angiotensin II acting inside the blood-brain barrier. PMID- 3044143 TI - Penetration of peripheral glucose and insulin into cerebrospinal fluid in rats. AB - In this study the penetration of plasma insulin and glucose into the cerebrospinal fluid (CSF) was investigated. Rats were implanted with cannulas in the cisterna magna and into the left and right jugular veins. Freely moving rats were intravenously infused during 4 h with either glucose solution (10 mg/min) or saline. Before, during, and after the infusions, simultaneous blood and CSF samples were taken. Infusion of glucose led to an immediate rise of both plasma glucose and insulin. Although CSF glucose followed plasma glucose within 10 min, CSF insulin was unchanged until 40 min. After termination of the glucose infusion, levels of all substances returned to base line within 10 min. Twenty four-hour food deprivation resulted in a significant decrease of plasma glucose, plasma insulin, CSF glucose, and CSF insulin. At the onset of eating after deprivation, an increase of plasma glucose and insulin occurred within 10 min, whereas CSF glucose was delayed between 10 and 40 min, after which ad libitum values were attained or surpassed. CSF insulin always remained below ad libitum levels. It is concluded that 1) glucose and insulin penetrate into the CSF and 2) CSF insulin and glucose can fulfill a putative feedback in homeostatic control of food intake and body weight. PMID- 3044145 TI - Thirst in Brattleboro rats. AB - Thirst mechanisms in Brattleboro rats are activated because of a deficiency in circulating vasopressin. Plasma osmolality, renin, and angiotensin II (ANG II) are increased. We measured the responsiveness of Brattleboro rats and appropriate control strains to cellular and extracellular thirst stimuli taking the spontaneous base-line water intake into account. Brattleboro rats drank more in response to intraperitoneal hypertonic NaCl than controls, but when their fluid losses were prevented by nephrectomy they did not overdrink. Despite low urinary concentration, Brattleboro rats excreted the sodium load at least as rapidly as the controls. Brattleboro rats drank after intracranial injection of renin, renin substrate, and ANG I and II. The dose-response curves were similar to controls, although the Nottingham Long-Evans control strain drank significantly less in response to some doses of the peptides. Intracranial captopril inhibited renin- and ANG I-induced but not ANG II-induced drinking. Isoproterenol reduced spontaneous drinking of Brattleboro rats but increased drinking in controls. However, when urinary losses were prevented by ureteric ligation, isoproterenol caused markedly greater water intake in Brattleboro rats than in controls. Subcutaneous captopril in moderate, thirst-enhancing doses also caused a larger increase in water intake in Brattleboro rats than in controls. Therefore the renin-angiotensin system of Brattleboro rats is more responsive to renin dependent thirst challenges than that of normal controls. PMID- 3044146 TI - Effect of teleost GnRH on reinitiation of oocyte meiosis in goldfish, in vitro. AB - The effects of native and an agonist analogue of teleost gonadotropin-releasing hormone (GnRH) [(D-Arg6, Trp7, Leu8, Pro9-NHEt)-GnRH; tGnRH-A] on the hormone induced meiotic maturation of goldfish oocytes were investigated. Incubation of follicle-enclosed goldfish oocytes with either carp gonadotropin (GtH) or meiogenic progestogens, including 17 alpha-hydroxyprogesterone (HP) and 17 alpha,20 beta-dihydroxyprogesterone (DHP), reinitiated oocyte meiosis in vitro, as indicated by germinal vesicle dissolution (GVD). Addition of tGnRH-A (10(-7) M) significantly attenuated the GVD response to GtH, HP, and DHP in a dose related fashion; tGnRH-A reduced the magnitude of progestogen-induced GVD response without significantly affecting the sensitivity of the oocytes to the meiogenic steroids. Similarly, administration of native teleost GnRH (tGnRH) significantly attenuated GtH-induced GVD response, although the 50% maximal response of the native peptide was significantly higher than that of tGnRH-A (380.9 +/- 125 and 2.72 +/- 0.94 nM, respectively). The present results provide the first demonstration of an inhibitory action of a GnRH agonist on the process of progestogen-induced oocyte meiosis and support the hypothesis that GnRH or a GnRH-like peptide might play a modulating role in the control of oocyte meiosis at the ovarian level in goldfish. PMID- 3044148 TI - Effect of starvation and food intake on sympathetic activity. AB - These studies have examined the effect of fasting and nutrient loads on sympathetic firing rate in three groups of rats that develop widely divergent degrees of obesity when eating a high-fat diet. Starvation of Sprague-Dawley rats for 24 or 48 h was associated with a decrease in basal sympathetic activity of nearly 25% in the first 24 h and of slightly greater than 30% in 48 h. This decline in sympathetic activity paralleled the loss of body weight and reduction in adipose tissue mass. After starvation for 48 h, Osborne-Mendel rats, which readily develop obesity when eating a high-fat diet, showed a greater decrease in basal sympathetic activity than did the diet-resistant S 5B/P1 rats. A single liquid 36-kcal intragastric meal was associated with an acute 30% increase in sympathetic firing rate in the overnight-fasted Sprague-Dawley rats. The values 3 h after the meal had returned halfway to normal, and by 6 h they were more than 85% of the way to normal. An intravenous injection of glucose produced a greater rise in sympathetic activity in diet-resistant S 5B/P1 rats than in the diet sensitive Osborne-Mendel rats. These data are consistent with the hypotheses that sympathetic activity is positively related to nutrient status, that it varies between strains of rats, and that it can be acutely increased by an intragastric meal or by intravenous glucose. PMID- 3044147 TI - Hormonal and electrolyte responses of conscious sheep to 96 h of hypoxia. AB - Hypoxia alters the relationship of aldosterone secretion to plasma renin activity. The potential role plasma electrolytes play in this modification is not clear. This study analyzed the interrelationships among renin, aldosterone, vasopressin (ADH), and plasma electrolytes during 96 h of normobaric hypoxia. Eight ewes were exposed, in discrete experiments, to hypocapnic hypoxia [arterial O2 tension (PaO2) 37-42 mmHg, arterial CO2 tension (PaCO2) 26-28 mmHg] and eucapnic hypoxia (PaO2 40-43 mmHg, PaCO2 28-31 mmHg) by N2 dilution in an environmental chamber. Urine output (24 h) was measured, and arterial plasma samples were collected during the normoxic control period and at 24-h intervals of hypoxia. Plasma Na+, K+, renin, and ADH levels did not change from the normoxic values during either hypocapnic or eucapnic hypoxia. However, urinary aldosterone excretion [critical significance (alpha) less than 0.046] and K+ excretion (alpha less than 0.046) decreased markedly during each type of hypoxia. All sheep developed a pronounced negative K+ balance by 96 h of hypoxia. These data suggest that plasma K+ concentration is preserved by movement of K+ out of the intracellular compartment; this change in K+ distribution may inhibit aldosterone secretion during hypoxia. PMID- 3044149 TI - The epidemiology of S. sonnei and S. flexneri in Pima County, Arizona: an exploratory study. AB - Investigation of 189 cases of shigellosis reported to the Pima County, Arizona Health Department in 1986 revealed that 23 per cent of cases could be attributed to travel to Mexico, and 10 per cent to day care attendees and their household contacts. No source of infection or high-risk activity could be demonstrated for 43 per cent of the cases. Households in which S. flexneri occurred were more likely to be characterized by crowded living situations and to have no known source of infection. PMID- 3044150 TI - Resistance to innovation: the case of the community health center. PMID- 3044151 TI - Human malaria transmission studies in the Anopheles punctulatus complex in Papua New Guinea: sporozoite rates, inoculation rates, and sporozoite densities. AB - Malaria sporozoite rates and inoculation rates were measured over periods up to 25 months in the different anopheline species biting humans in 13 villages in Madang Province, Papua New Guinea. Analysis of three members of the Anopheles punctulatus complex, 68,458 An. farauti, 36,779 An. koliensis, and 11,667 An. punctulatus caught in landing catches was made using monoclonal antibody based ELISAs to detect sporozoites of Plasmodium falciparum and P. vivax. Sporozoite rates ranged from 0%-5.5% in An. farauti, 0.2%-3.8% in An. koliensis, and 0%-3.3% in An. punctulatus. In addition, over 3,000 An. longirostris were analyzed and sporozoites were not detected in this species. No significant differences were observed between the three vector species in the densities of P. falciparum sporozoites (geometric mean 2,320). However, the geometric mean P. vivax sporozoite density was significantly higher in An. punctulatus (350) than in either An. koliensis (160) or An. farauti (150). An. koliensis was less susceptible to infections of P. falciparum or P. vivax than either An. farauti or An. punctulatus. Variations in average sporozoite and inoculation rates were found among different villages, despite their close geographic proximity. Sporozoite and inoculation rates varied greatly within a village over time, but malaria transmission was perennial with a higher transmission during the wet season by An. koliensis and An. punctulatus. PMID- 3044152 TI - Use of avidin-biotin-glucose oxidase complex to detect antimalarial antibody in serum by light microscopy. AB - An immunohistochemical assay was developed combining an avidin-biotin-glucose oxidase complex procedure (ABC-GO) with light microscopy to detect specific antibody against Plasmodium falciparum. Thin blood films were prepared from culture material of P. falciparum and fixed with acetone. Antibody was detected by successive incubations with test serum, biotinylated goat antihuman antibody, avidin-biotin-glucose oxidase complex, and glucose oxidase substrate. In the presence of reactive serum, a blue precipitate formed on the parasites and could be visually observed with a 40x objective. Sera from patients with single infections for P. vivax or P. ovale were unreactive. No cross-reactivity was observed with sera from patients with rheumatoid arthritis, filariasis, amebiasis, schistosomiasis, dengue, scrub typhus, leptospirosis, or toxoplasmosis. The sensitivity of ABC-GO is comparable to that of the indirect fluorescent antibody test. PMID- 3044153 TI - Natural antibodies against three distinct and defined antigens of Plasmodium falciparum in residents of a mesoendemic area in Gabon. AB - The magnitude of the antibody response to three distinct and defined antigens of Plasmodium falciparum was assessed in 144 inhabitants of the Kassa district (Haut Ogooue Province, Gabon), a region where malaria is mesoendemic. Antibodies against a polypeptide consisting of 40 (Asn-Ala-Asn-Pro) repeats of P. falciparum circumsporozoite protein [(NANP)40] were detected by ELISA. Antibodies against the fusion peptide 31.1, which consists of the N-terminal portion of the 190-200 kDa glycoprotein, were also detected by ELISA. Antibodies against ring-infected erythrocyte surface antigens (RESA), mainly the P. falciparum 155 kDa antigen (Pf 155), were detected by IFA on glutaraldehyde-fixed P. falciparum infected red blood cells (IRBC). In addition, a standard IFA employing air-dried P. falciparum IRBC was used to detect antibodies against intraerythrocytic asexual forms. Parasitemia and spleen enlargement were also recorded. The frequency of sera positive for specific antibodies increased with age for all the antigens tested. Plateau antibody levels were reached at different ages for the different antigens. Individual antibody responses varied in titer to the different antigens. Subjects with parasite-negative thick smears showed higher titers of anti-31.1 as well as an increased frequency of anti-RESA antibodies compared to subjects having positive smears. No differences in the titer and in the prevalence of anti-(NANP)40 antibodies were found between these groups. The results suggest that the antibody response against asexual blood stage antigens, especially anti-RESA and anti-31.1, may play a role in controlling parasitemia. PMID- 3044154 TI - X-ray microanalysis of Plasmodium falciparum and infected red blood cells: effects of qinghaosu and chloroquine on potassium, sodium, and phosphorus composition. AB - Cryosections of human red blood cells infected by Plasmodium falciparum were analyzed by energy dispersive x-ray microanalysis to determine the elemental composition of the parasites and their red cell hosts separately. The effects of two antimalarial drugs, qinghaosu and chloroquine, on potassium, sodium, and phosphorus concentrations were studied. Malarial infection causes a decrease in potassium concentration and an increase in sodium concentration in the host red cells. The drastic change in the cation composition, however, occurs only in red cells infected by late stage parasites (late trophozoite and schizont). Red cells infected by early stage parasites (ring stage) show only small changes in sodium concentration. Furthermore, the noninfected red cells in parasitized cultures show no difference in composition from those of normal red cells. Treatment of the parasitized cultures with qinghaosu (10(-6) M) or chloroquine (10(-6) M) for 8 hr causes phosphorus concentration of both early and late parasites to decrease. An 8 hr treatment with qinghaosu also produces a reduction in potassium and an increase in sodium concentrations in early and late parasites. In contrast, 8 hr treatment with chloroquine only causes a change in the sodium and potassium concentrations of the late stage parasites and does not affect the early stage parasites. PMID- 3044155 TI - Patterns of in vitro resistance to chloroquine, quinine, and mefloquine of Plasmodium falciparum in Cameroon, 1985-1986. AB - The drug sensitivity of 246 Plasmodium falciparum isolates was studied in vitro in five areas of Cameroon at the end of 1985. Results demonstrate that parasites resistant either to chloroquine, quinine, or mefloquine, or to two of these drugs, were prevalent in four of the areas investigated, but the drug response pattern varies widely from one area to another. The recent explosive emergence of chloroquine resistance in the south of the country, where both prevalences and levels are very high (up to 86%), contrasts with only moderate levels of resistance in the north. This may be related to differences in transmission by mosquitoes between Sahel and forest areas. Quinine resistance was observed in 24% of the isolates studied in vitro and was frequently associated with chloroquine resistance. The presence of isolates responding poorly to mefloquine, observed mainly in northern Cameroon, suggests that resistance may occur in areas where the drug has never been used. PMID- 3044156 TI - Analysis and preparation of Bartonella bacilliformis antigens. AB - Twenty-four antigens of Bartonella bacilliformis, a bacterium which causes bartonellosis in residents of high altitude valleys of the Andes, were identified by immunoblot and immunoprecipitation using rabbit anti-Bartonella sera as well as sera of patients. The antigens were designated according to their relative molecular mass which ranged from 16 to 160 kDa. Twelve antigens were detected by antibodies in sera of bartonellosis patients using immunoblot, of which six antigens were detected by immunoprecipitation. Antigens 25, 46, 65, 75, 99, and 160 were identified as probable cell wall antigens. Antigens 50, 65, and 75 detected long-persisting antibodies. Crude Bartonella antigen applied to ELISA reacted with anti-Chlamydia psittaci antibody as well as with antibody of unknown identity in human sera, whereas immunoblot and immunoprecipitation with Triton soluble antigens revealed Bartonella-specific results. Seven Bartonella antigens were prepared by high performance liquid chromatography of which one antigen (48 kDa) reacted Bartonella-specific when applied to ELISA. It was concluded that specificity of antibody determination with crude Bartonella antigen should be confirmed by either immunoblot or immunoprecipitation. PMID- 3044157 TI - Serological patterns and specificity in American cutaneous leishmaniasis. AB - Sera from diverse clinical forms of American cutaneous leishmaniasis show remarkably different patterns of reactivity to Leishmania mexicana and L. braziliensis after absorption with these two species of Leishmania. The enhanced species specificity of absorbed sera was confirmed by reactions with well characterized reference strains. The use of pairs of enriched polyclonal sera from mucocutaneous and diffuse cutaneous leishmaniasis permits the preliminary classification of isolates in the L. mexicana or L. braziliensis complexes by a simple, inexpensive, and quantitative immunoassay. PMID- 3044158 TI - [Therapeutic propositions in pains of the Pancoast and Tobias syndrome: review of the literature]. PMID- 3044160 TI - The treatment of acute polyradiculoneuritis with respiratory paralysis. AB - This paper reports 504 cases admitted with acute polyradiculoneuritis (AP) to Beijing Children's Hospital from 1975 through 1984. 343 of the 504 cases (68.1%) with AP had respiratory paralysis and in 198/504 (39.3%), tracheotomy was performed. In this study, none of the patients received steroids. We attempted to assess the grades of respiratory paralysis and established the criteria for tracheotomy. We also must perform the following procedures: 1) Artificial ventilation must be used correctly, 2) The secretion should be aspirated regularly in order to maintain a clear airway, 3) Sterilization must be strictly conducted to prevent cross infection. As a result, of 504 cases with AP 9 died, the mortality rate being 1.79%. The study suggests tracheotomy should be performed early and it is one of the most important procedures in the treatment of AP with respiratory paralysis. PMID- 3044159 TI - The "lost autoregulation hypothesis" and brain lesions in the newborn--an update. AB - Autoregulation of cerebral blood flow is an essential homeostatic mechanism which is easily disturbed during the stresses in the birth process. In its absence, moderate hypotension, a frequent complication in neonatal asphyxia, may induce cerebral ischemia, the arterial watershed zones in periventricular regions being particularly prone. The initial ischemia in perinatal stress produces functional disturbance (EEG depression), which is not readily reversible as the circulation improves. Hypotension, cerebral hypoperfusion and EEG depression precedes severe periventricular hemorrhage, which seems to be triggered by fluctuations in arterial blood pressure and flow, with fluctuations in the transmural pressure gradient across the capillary wall. The penetration of the hemorrhaging into surrounding brain tissue is related to the magnitude of the preceding ischemic insult. PMID- 3044161 TI - Paternal alcohol exposure affects offspring behavior but not body or organ weights in mice. AB - Male mice consumed liquid alcohol diets containing 20, 10, or 0% ethanol-derived calories (EDC) for 56-61 days. Animals fed the 10 and 0% EDC diets were pair-fed to 20% EDC animals. A nontreated ad libitum group was included to assess the effects of pair-feeding. After treatment, males were bred to nontreated females. Litter size, birthweight, bodyweight at 21 or 55 days of age, and organ weights except for thymus were not affected by paternal alcohol ingestion. There was a dose-related decrease in activity at 20 and 24 days of age in an activity chamber and a dose-related decrease in serum testosterone levels at 55 days of age in male offspring sired by alcohol-consuming males. Offspring sired by males consuming the 20% EDC diet also required fewer trials to learn a passive avoidance task and had longer latencies to reach the choice point in a T maze. PMID- 3044162 TI - Altered acylation of erythrocyte phospholipids in alcoholism. AB - The composition and metabolism of erythrocyte lipids were studied in 10 chronic alcoholic patients within 48 hr after discontinuation of alcohol intake and in 10 nonalcoholic control subjects. Chronic alcoholism produced no change in contents of cholesterol, total phospholipids, and proportions of phosphatidylcholine and phosphatidylethanolamine in erythrocyte phospholipids. The mean values of the rates of acylation of phosphatidylcholine and phosphatidylethanolamine with oleic acid were increased respectively by 59% (p less than 0.001) and 38% (p less than 0.05) as compared with the controls. There was no correlation between acylation rates and mean cellular volumes. Increases in acylation rates normalized over several weeks after alcohol withdrawal and were not related to a direct effect of alcohol on the intact erythrocyte, suggesting that these alterations result from ethanol-induced changes in the membrane during erythrocyte formation. The increased rates of acylation might modify the remodeling of the lipid matrix and thereby the membrane function. PMID- 3044164 TI - Alcohol use and depression symptoms among female nursing students. AB - A sample of 286 female nursing students responded to questions concerning drinking practices and symptoms of depression. After controlling for the possibly confounding effects of nursing student experiences (aspects of burnout such as depersonalization and social isolation) and the sociodemographic characteristics of the students, increased quantity of alcohol consumed per drinking occasion was found to be associated with increased symptoms of depression in the sober state. PMID- 3044163 TI - Use of an objective clinical scale in the assessment and management of alcohol withdrawal in a large general hospital. AB - A modified version of the Clinical Institute Withdrawal Assessment Scale (CIWA) was used in the management of alcohol withdrawal in a general hospital. Patients who developed seizures or confusion were noted to score higher on the scale, even before these complications, than patients who remained uncomplicated (21.7 +/- 1.2 compared to 15.6 +/- 0.55). When the score was used as a guide for treatment, it was found that patients scoring greater than 15 were at significantly increased risk of severe alcohol withdrawal if they remained untreated (RR, 3.72; 95% confidence interval, 2.85-4.85). The higher the score the greater this relative risk. Some patients however, still suffered complicated withdrawals although their scores were low or they were apparently adequately treated. It is concluded that the use of an objective clinical scale of alcohol withdrawal is valuable in a general hospital to identify those patients in early withdrawal who need sedation to avoid complication. There will however, be a small group of patients whose clinical course will be difficult to predict and further work is needed to determine the reasons for this. PMID- 3044165 TI - Event-related brain potentials in individuals at high and low risk for developing alcoholism: failure to replicate. AB - Event-related brain potentials (ERPs) were used to compare young men having a positive paternal family history for alcoholism (FHP) with carefully matched control subjects having no family history for alcoholism (FHN). The P300 ERP component was obtained from all subjects (n = 10/group) with a complex auditory paradigm before and on two occasions after they received a placebo drink which they were told might contain alcohol. The procedures employed replicated those of a previous study in which FHP subjects showed diminished P300 potentials compared to FHN subjects under the placebo as well as ethanol consumption conditions--a finding which raised the possibility that the P300 ERP component might be a biological marker for subjects at high risk to develop alcoholism. No differences between the family history groups were obtained for the P300 or any other ERP component using the replication procedures. Both groups demonstrated a decrease in P300 amplitude across trial blocks in a similar fashion suggesting that habituation effects may have diminished the ERP response. PMID- 3044166 TI - Impact of depressive symptomatology on alcohol problems in women. AB - The association between the amount of depression manifest at time of an alcoholic woman's admission to treatment, the course of alcoholism and its consequences, and early psychosocial vulnerabilities was examined in a sample of women in treatment for alcoholism (n = 301). Two indices of depression, one reflecting low self-esteem and the other current mood were constructed by factor analysis of interview items measuring depression (drawn from Center for Epidemiological Studies Depression scale, Diagnostic Interview Scale, and Personal Attributes Questionnaire). Both depression indices were associated with earlier age of onset and earlier loss of control. Binge drinking was associated with current mood, while daily/weekend drinking was associated with low self-esteem. Both measures were associated with the consequences of alcoholism. An examination of alcoholic women's background revealed that there are some influences in the early family environment that are associated with increased levels of depression in alcoholic women. PMID- 3044167 TI - Manipulation of alcohol preference in C57BL/6J mice by episodes of food poisoning. AB - Individual differences in the amount of alcohol consumed in a choice situation are found in highly inbred C57BL/6J mice. The extent to which environmental stress can modify alcohol preference was studied by coupling acute episodes of poisoning with restricted fluid availability, and recovery with free choice of drinking fluids. Addition of actinomycin D, a mycotoxin, to ordinary chow during 2-day periods produced acute episodes of nonlethal food poisoning from which the mice recovered rapidly. Consumption of a 10% alcohol solution and of water was recorded for several weeks before poisoning and for several weeks after the last episode. By varying drinking fluids available to the mice during the episodes of poisoning, long-lasting changes in alcohol preference were produced. When 10% alcohol was the sole drinking fluid available during poisoning, preference for the alcohol solution was abolished. When water was the sole fluid during poisoning, alcohol preference was increased above the already high levels established in the baseline and above a control group that was restricted to water during the treatment periods but was not poisoned. This increased alcohol preference was due to a nearly complete suppression of water intake in the posttreatment period; there was no significant increase in amount of alcohol consumed. The greatest individual differences in subsequent alcohol preference were found in the group of mice which continued to have free choice of alcohol and water during episodes of poisoning. The variety of responses to the same treatment show how environmental influences outside the experimenter's control may account for the variability found in voluntary alcohol consumption among genetically homogeneous mice. PMID- 3044168 TI - Effects of ethanol on mature offspring of mice given ethanol during pregnancy. AB - Male offspring of mice maintained on isocaloric liquid diets containing either 20% ethanol or sucrose derived calories during pregnancy were tested on their ability to discriminate different ethanol doses as adults. They were trained to lever-press for a food reward on each of two levers in an operant chamber, and were then maintained on an FR20 reinforcement schedule. After response rates stabilized, ethanol discrimination training was initiated by reinforcing only responses made on the drug appropriate-lever after i.p. injections of ethanol (1.0 g/kg) or water. After learning to discriminate the 1.0 g dose, the animals' ability to discriminate doses of 0.25, 0.50, 0.75, 1.0, and 1.5 g/kg was assessed by determining the percentage of responses made on the drug lever during a 2-min test period. Compared to sucrose controls, mice exposed to alcohol in utero learned the lever response more slowly and were less responsive to injected ethanol as evidenced by a reduced effect of the drug on response rates and by a reduction in their ability to discriminate the presence of injected ethanol. The results indicate that prenatal ethanol exposure can have long term consequences which reduce the effects of ethanol in fully mature animals. PMID- 3044169 TI - The Preoccupation with Alcohol Scale: development and validation. AB - The purpose of the present study was to develop a psychometrically sound measure of preoccupation with alcohol in a group of young, heavy drinking men and to examine some correlates of this drinking style. A brief Preoccupation with Alcohol Scale (PAS) was derived that exhibited substantial internal consistency. Data from a sample of the 67 males indicated that higher PAS scores were significantly related to alcohol consumption and frequency of intoxication within many different contextual circumstances. The PAS was also related to alcohol problems after controlling for average daily alcohol consumption and frequency of intoxication, suggesting that scores on the PAS are related to, but not redundant with, level of alcohol consumption. Results from a cross-validation sample provided additional support for these findings. The authors suggest that a preoccupation with alcohol may be a cognitive-behavioral risk factor for the development of severe alcohol problems and alcoholism. PMID- 3044170 TI - Heavy drinking and regular psychoactive drug use among gynecological outpatients. AB - Self-reported patterns of alcohol and drug use were examined in gynecological outpatients (n = 1967). Seventeen percent were heavy drinkers, 14% regularly used psychoactive drugs with the potential for nonmedical use, and over 3% both drank heavily and regularly used psychoactive drugs. Significantly higher alcohol intakes (mean usual number of drinks per occasion was 3.6 compared to 3.1, p less than 0.01) and more heavy drinkers (26% compared to 19%, based on quantity frequency, p less than 0.01) were associated with regular psychoactive drug use. However, the relationship between other measures of alcohol use and regular psychoactive drug use varied as a function of age, marital status, and employment status. Regular psychoactive drug use was significantly elevated among outpatients who were in their fifties, retired/other (not housewives, employed, or looking for work, e.g. students or disabled), not married, or white. The risk of regular psychoactive drug use was more than 1.5 times higher among heavy drinkers than among nonheavy drinkers. It is estimated that there are over 35 million gynecologic office visits per year; therefore, increased case-finding in this population has the potential to reach large numbers of women at risk who otherwise might not be identified. PMID- 3044171 TI - Roles of hormonal and nutritional factors in the regulation of rat liver alcohol dehydrogenase activity and ethanol elimination rate in vivo. AB - Fasting reduced the liver alcohol dehydrogenase (ADH) activity by 51% (p less than 0.001). Insulin, within 2 hr, increased the ADH activity found in fasted animals by 28% (p less than 0.02). Insulin administration failed to stimulate the reduced ADH activity in diabetic rats. However, ADH activity in the diabetic-fed rats decreased by 52-54% (p less than 0.001) compared to normal-fed rats regardless of whether they were meal-fed or refed the normal chow. Glucagon blocked by 15% (p less than 0.02) the increase in ADH activity associated with refeeding. Furthermore, insulin caused a marginal stimulation of ethanol elimination rate (EER) when administered to fasted rats. All these results imply that insulin and glucagon may not be the only determining factors in the control of liver ADH activity associated with fasting and refeeding. Meal-feeding or refeeding a high carbohydrate fat-free diet compared to the normal chow-diet caused 29% (p less than 0.001) and 36% (p less than 0.05) decreases in ADH activity, respectively. Concomitant decreases in EER caused by high carbohydrate fat-free diet feeding were also observed under identical conditions. These results raise the possibility that the amount and the type of carbohydrate may be crucial in the regulation of ADH and EER. Alternatively, the presence of fat may be important in maintaining the normal level of ADH and EER. PMID- 3044172 TI - Role of alcohol-induced hypothermia in mediating the teratogenic effects of alcohol in C57BL/6J mice. AB - The purpose of this study was to determine the role of alcohol-induced maternal hypothermia in the teratogenic actions of alcohol. C57BL/6J mice were administered an acute dose of alcohol (5.8 g/kg orally) or isocaloric sucrose on day 10 of gestation. One half of each group was placed for 6 hr in an incubator set at 32 degrees C and the other half was housed in the incubator at room temperature (22 degrees C). As expected, acute prenatal alcohol exposure at this time of gestation was associated with decreased birth weight and an increase in limb and kidney malformations. The significant alcohol x environmental temperature interaction on these dependent variables indicated that the teratogenic insult was not attenuated, but was in fact even greater for the 32 degrees C/alcohol group. An absence of a main effect of environmental temperature indicated that the 32 degrees C environment, per se, was not teratogenic. Thus, maternal hypothermia is probably not an etiological factor in animal models of fetal alcohol syndrome. Moreover, antagonism of alcohol-induced maternal hypothermia exacerbates the teratogenic actions of alcohol observed at room temperature. PMID- 3044173 TI - Neonatal and maternal hair zinc levels in a nonhuman primate model of the fetal alcohol syndrome. AB - The role of zinc deficiency in the etiology of ethanol-associated fetotoxicity was assessed by measurement of maternal and newborn hair zinc content in a nonhuman primate model of the fetal alcohol syndrome. The model best approximates the human situation for length of gestation and type of placentation, coupled with the ability to control for nutritional factors. All mothers received 110% of their minimum daily caloric requirements as a balanced, nutritionally complete diet, including a minimum of 3.5 mg zinc per day. Over a 2-year period, maternal hair samples from 17 pregnancies (using 12 females) were obtained at term. There were 16 live, full-term neonates (nine ethanol exposed and seven control) from whom samples were taken within 1 hr of birth. The ethanol-exposed infant monkeys had a significantly higher incidence of craniofacial dysmorphology and developmental delay compared to the controls. There was no difference in hair zinc levels between ethanol-exposed and control animals for either the mothers or the newborns. Neonatal levels were, however, consistently higher than corresponding maternal. Although the findings cannot exclude transient or early gestational zinc deficiency as a factor, they provide further evidence that ethanol (and/or its metabolites) is the proximate toxin in the type of fetal injury seen in the fetal alcohol syndrome. PMID- 3044174 TI - Changes in erythrocyte enzyme activities during erythrocyte aging in alcoholism. AB - Aldehyde dehydrogenase, glucose-6-phosphate dehydrogenase, and pyruvate kinase activities were determined in erythrocytes of various ages, separated by Percoll gradient centrifugation, in 13 alcoholic patients and eight control subjects. The total erythrocyte activities of all three enzymes were not affected by alcoholism, however, the youngest cells of alcoholics had a decreased aldehyde dehydrogenase activity, while both glucose-6-phosphate dehydrogenase and pyruvate kinase activities were increased. The depression of aldehyde dehydrogenase activity not only persisted, but became more marked after 2 weeks of abstinence, while the enhanced activities of the two other enzymes returned to normal. These observations suggest that chronic alcohol ingestion suppresses aldehyde dehydrogenase in the bone marrow, while it enhances other erythrocytic enzymes. PMID- 3044175 TI - Carbohydrate-deficient transferrin, a marker for chronic alcohol consumption in different ethnic populations. AB - Serum levels of carbohydrate-deficient transferrin (CDT) were determined in a racially mixed population of 107 alcoholics, 18 healthy, nonalcoholic control subjects, 62 abstinent alcoholics, and in 64 Caucasian patients with various nonalcoholic liver diseases. The upper limit of normal CDT levels was 80 mg/liter (2 SD above the mean). CDT values exceeding this level were found in more than 80% of Black, Puerto Rican, and Caucasian alcoholics who had consumed greater than or equal to 50 g of alcohol/day for 1 month or longer prior to testing. Puerto Rican alcoholics had higher CDT values than the Black and Caucasian ethnic groups; however, these differences were significant only when compared to the Black population. Of 64 patients with nonalcoholic liver diseases, one individual with chronic active hepatitis (CAH) with an alcohol consumption of 20 g/day, and 10 of 26 subjects with primary biliary cirrhoses (PBC), who claimed to consume either no or only occasional moderate amounts of alcohol, had CDT levels ranging from 81 to 144 mg/liter. Seven of these individuals were in advanced stages of PBC. Total transferrin levels were variable and not significantly different in all subject groups studied. CDT/total transferrin ratios were increased in most patients with abnormal amounts of CDT, and there was a significant correlation between these ratios and CDT levels in all study groups. Serum enzyme parameters as well as red blood cell mean corpuscular volumes did not correlate with CDT values.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3044176 TI - Time course and reversibility of ethanol's suppressive effects on axon sprouting in the dentate gyrus of the adult rat. AB - Ethanol was administered chronically to adult rats in a liquid diet for 14 days preceding and for 5, 7, 8, 9, or 10 days following the unilateral destruction of the entorhinal cortex. Control groups received a diet of lab chow and water and were sacrificed at comparable survival times. An additional experimental group was given ethanol until 9 days after the lesion, then switched to lab chow and water and sacrificed 1 day later. Coronal sections through the dorsal hippocampal formation were stained and analyzed histochemically for the localization of acetylcholinesterase (AChE). Quantitative measurements of the histochemical patterns in the molecular layer of the dentate gyrus were obtained. Ethanol exposure inhibited the withdrawal of the acetylcholinesterase-stained septohippocampal fibers and limited the typical lesion-induced expansion of the pale-staining commissural/associational zone in the molecular layer of the denervated dentate gyrus. However, abstinence from ethanol for just 24 h released the inhibitory effect on the acetylcholinesterase-staining fibers, resulting in a significant expansion of the commissural/associational zone. PMID- 3044177 TI - Effect of prenatal exposure to ethanol on the development of cerebral cortex: I. Neuronal generation. AB - Prenatal exposure to ethanol causes profound disruptions in the development of the cerebral cortex. Therefore, the effect of in utero ethanol exposure on the generation of neurons was determined. Pregnant rats were fed a liquid diet in which ethanol constituted 37.5% of the total caloric content (Et) or pair-fed an isocaloric control diet (Ct) from gestational day (GD) 6 to the day of birth. The time of origin of cortical neurons was determined in the mature pups of females injected with [3H]thymidine on one day during the period from GD 10 to the day of birth. The brains were processed by standard autoradiographic techniques. Ethanol exposure produced multiple defects in neuronal ontogeny. The period of generation was 1-2 days later for Et-treated rats than for rats exposed prenatally to either control diet. Moreover, the generation period was 1-2 days longer in Et-treated rats. The numbers of neurons generated on a specific day was altered; from GD 12 19 significantly fewer neurons were generated in Et-treated rats than in Ct treated rats, whereas after GD 19 more neurons were born. The distribution of neurons generated on a specific day was disrupted; most notable was the distribution of late-generated neurons in deep cortex of Et-treated rats rather than in superficial cortex as they are in controls. Cortical neurons in Et treated rats tended to be smaller than in Ct-treated rats, particularly early generated neurons in deep cortex. The late-generated neurons in Et-treated rats were of similar size to those in Ct-treated rats despite their abnormal position in deep cortex. Neurons in Ct-treated rats tended to be rounder than those in Et treated rats which were more polarized in the radial orientation. A proliferation index, which was based on the amount of autoradiographic signal over each labeled neuron, indicated that an additional, late surge in proliferative activity occurred in Et-treated rats on GD 20-21. The amount of [3H]thymidine incorporated each day was determined by biochemical analyses. In Ct-treated rats, incorporation increased to a maximum on GD 17 and decreased thereafter. In Et treated rats, there were two maxima, the first on GD 18 and the second on GD 20. These data fully support the findings of the autoradiographic analyses. The present data show that neuronal generation is profoundly affected by ethanol. Such disturbances result from ethanol-induced abnormalities in neuronal proliferation and migration. PMID- 3044179 TI - Alcohol and aldehyde dehydrogenase isoenzymes in Sioux North American Indians. PMID- 3044178 TI - Transferrin phenotype and level of carbohydrate-deficient transferrin in healthy individuals. AB - Elevated concentrations of carbohydrate-deficient components of transferrin (CDT) in serum may be used as a sensitive and specific marker of regular, high alcohol consumption. When determined by a new, simplified assay, CDT values are nearly normally distributed in low- or non-alcohol-consuming control populations. The importance of transferrin phenotype for this normal variation was analyzed in 100 healthy, European men and women with no or negligible alcohol intake. No significant relation was found between phenotype and CDT value in this population. The three rare B-variants found had low CDT levels, and one subject, examined outside the study, with a rare D-variant indicated that D-variants may result in false-positive CDT values. Moreover, women tended to have somewhat higher values than men, in whom CDT levels were weakly correlated with age. Other as yet undefined biological factors are clearly responsible for the major part of the normal variation of CDT values in nonalcoholic individuals. PMID- 3044180 TI - [Histogenesis and oncogenes of breast cancer]. PMID- 3044181 TI - [Nalbuphine in comparison with piritramid and placebo in postoperative pain therapy following intubation anesthesia with halothane. Side effects and effectiveness]. AB - The aim of the study was a comparison of the side-effects and efficacy of nalbuphine, piritramide, and placebo in patients during recovery from halothane anesthesia. METHODS: Neurosurgical (vertebral surgery) and otolaryngological patients (surgery of face and neck) were operated under halothane anesthesia. Postoperatively 20 patients received 20 mg nalbuphine, 21 patients 15 mg piritramide, and 19 patients 0.9% NaCl for pain therapy in a randomized and double-blind manner. Respiratory function was monitored by blood gas analysis, hemodynamic function by noninvasive measurements. The analgetic and sedative effects were estimated by the patients (visual analog scale) and the investigator (4-point scale). If the treatment was ineffective, the study was interrupted and a known analgesic was prescribed. RESULTS: The noninvasively measured hemodynamic parameters were unchanged. On the other hand, in the nalbuphine group mean arterial pCO2 increased significantly (max. 55.4 mmHg after 20 min), over the piritramide group (max. 51.2 mmHg before treatment) and the placebo group (max. 55.1 mmHg before treatment). Drowsiness, in 8 patients in each of the treatment groups and 3 patients in the placebo group, was the most frequent side-effect. After nalbuphine the pain threshold was significantly higher than after treatment with piritramide and placebo. The study was interrupted because of inefficacy in no patients from the nalbuphine group, 2 patients from the piritramide group, and 6 patients from the placebo group. There were no differences in the sedative effects. CONCLUSIONS: Nalbuphine seems to have better analgesic effects then piritramide. Both cause no hemodynamic alterations.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3044182 TI - [Experiences with parenteral administration of diltiazem in coronary surgery patients]. AB - Diltiazem is a calcium channel blocker whose effects lie between those of the two other important calcium antagonists nifedipine and verapamil. In addition to vasodilation, it has a negative dromotropic effect with prolongation of the A-V interval. In animal experiments and human investigations, diltiazem improves the function of ischemic myocardium due to a direct dilating effect on coronary vessels. The purpose of the present study was to investigate the hemodynamic effects of diltiazem in patients before and during coronary revascularization. METHODS. The study included 60 consenting male patients with coronary heart disease. Twenty premedicated patients randomly received 0.3 mg/kg diltiazem or placebo within 3 min before induction of anesthesia. Hemodynamic measurements (arterial pressure, heart rate, mean pulmonary arterial pressure, pulmonary capillary pressure, right atrial pressure and cardiac output) were taken during the following 21 min. Before cannulation of the great vessels for institution of extracorporeal circulation (ECC), 20 other patients received 0.014 mg diltiazem or placebo/kg per min over 20 min. In addition to the above mentioned hemodynamic measurements, left ventricular parameters (LVP, LVEDP, dp/dt) were directly registered, and 5 min after the end of ECC the measurements were repeated with the same preload as before the ECC. Twenty additional patients received 0.014 mg diltiazem or placebo/kg per min within 21 min during ECC observing arterial perfusion pressure and oxygenator volume. RESULTS. Pre- and intraoperatively diltiazem caused a decrease in mean arterial pressure; cardiac index increased only during the preoperative investigation period (Tables 1, 2), whereas stroke volume index increased pre- and intraoperatively; heart rate decreased in all patients as well as dp/dt (Fig. 1).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3044183 TI - The avidin-biotin complex in bioanalytical applications. PMID- 3044184 TI - The seed stock concept and quality control for cell lines. AB - Procedures for the characterization of cell culture lines are reviewed. It is recommended that seed stocks be developed and that major quality-control efforts be applied first to those holdings, with essential tests repeated on cultures from working or distribution stocks prepared subsequently. Steps are taken throughout to ensure absence of microbial contaminants. Mycoplasma are especially troublesome. Investigators need also to be aware of potential endogenous viruses which may be hazardous or interfere with interpretation of results. Cell line species and absolute identities are established by fluorescent antibody staining, by determination of isoenzyme profiles, and by cytogenetic analyses. Additional characterizations will vary with cell type but may include tests for specific products, cytoskeletal proteins, tissue specific antigens, or immunoglobulins. Credentials of lines added to a collection plus representative cultures can be returned to the originators for concurrence before distribution. Advisory committees may be established as circumstance dictates to provide additional expertise regarding quality control for the specific collection being developed. PMID- 3044185 TI - Formation and characterization of precise eucaryotic transcription complexes using a semisynthetic DNA template and specific oligoribonucleotide primers. AB - An artificial template of defined sequence which supports specific in vitro initiation and elongation by yeast RNA polymerase II has been constructed. This template is a pBR322 derivative which contains a synthetic oligonucleotide inserted into the BamHI cloning site. The sequence of this oligonucleotide is such that when the plasmid is restricted with SacI the two ends obtained are identical. The addition of an oligodeoxycytidylate chain to the 3' hydroxy termini produces a DNA template, (poly dC-p+22), with the sequence: 3'(C)nTCGA GAGTCTCCTA. . . . The underlined position denotes the beginning of the duplex region. When initiation is primed with the diribonucleotide GpC the predicted sequence of the transcript obtained is: 5'GCUCUCAGAGGAU. . . . Kinetic and product analyses indicate that a ternary complex containing a precise length of transcript can be produced which is subsequently resistant to heparin inactivation. Initiation can also be directed to a specific position dictated by a tri or tetraribonucleotide primer. PMID- 3044186 TI - A malachite green procedure for orthophosphate determination and its use in alkaline phosphatase-based enzyme immunoassay. AB - An improved procedure for phosphate determination based on a highly colored complex of phosphomolybdate and malachite green is described. All necessary reagents are combined in one concentrated solution, making the assay sensitive and convenient. The procedure is based on the finding that the dye is easily soluble and stable in the presence of 6 N acid. The addition of Tween 20 is required to stabilize the dye-phosphomolybdate complex at phosphate concentrations above 10 microM. The time of color development at 25 degrees C is about 3 min. The procedure was adopted to measure alkaline phosphate activity in heterogeneous enzyme immunoassay with rho-nitrophenyl phosphate and pyrophosphate as substrates. In both cases, a 4-fold increase in sensitivity in terms of absorbance readings was obtained compared to the standard method based on rho nitrophenol measurement. In visual analysis, the gain in sensitivity was as high as 20-fold, due to contrast color change (yellow to greenish blue). PMID- 3044187 TI - Laser-excited time-resolved solid-phase fluoroimmunoassays with the new europium chelate 4,7-bis(chlorosulfophenyl)-1,10-phenanthroline-2,9-dicarboxylic acid as label. PMID- 3044188 TI - Circulatory effects of weaning from mechanical ventilation: the importance of transdiaphragmatic pressure. PMID- 3044189 TI - Acute left ventricular dysfunction during unsuccessful weaning from mechanical ventilation. AB - The authors studied the hemodynamic effects of rapidly weaning from mechanical ventilation (MV) 15 patients with severe chronic obstructive pulmonary disease (COPD) and cardiovascular disease who were recovering from acute cardiopulmonary decompensation. In each patient, 10 min of spontaneous ventilation (SV) with supplemental oxygen resulted in reducing the mean esophageal pressure (X +/- SD, + 5 +/- 3 to -2 +/- 2.5 mmHg, P less than .01) and increasing cardiac index (CI) 3.2 +/- 0.9 to 4.3 +/- 1.3 1/min/M2, P less than .001), systemic blood pressure (BP 77 +/- 12 to 90 +/- 11 mmHg, P less than .001), heart rate (HR 97 +/- 12 to 112 +/- 16 beats/min, P less than .001), and, most importantly, transmural pulmonary artery occlusion pressure markedly increased (PAOPtm 8 +/- 5 to 25 +/- 13 mmHg, P less than .001), mandating a reinstitution of MV. In four patients with left ventricular (LV) catheters, the PAOP correlated with the LV end diastolic pressure during both MV and SV. Gated blood pool imaging showed SV increased the LV end-diastolic volume index (65 +/- 24 to 83 +/- 32/M2, P less than .002) with LV ejection fraction unchanged. Patients were treated for a mean of 10 days with diuretics, resulting in a reduction of blood volume (4.55 +/- 0.9 1 to 3.56 +/- 0.55 1) and body weight (-5 kg, P less than .001). Subsequently, nine of the 15 patients were weaned successfully from mechanical ventilation with unchanged PAOP. PMID- 3044190 TI - The introduction of hedonal: a Russian contribution to intravenous anesthesia. PMID- 3044191 TI - [Effect of artificial circulation with pulsatile and non-pulsatile flow on indicators of carbohydrate metabolism in patients with acquired heart defects]. PMID- 3044192 TI - [Effect of long-term artificial ventilation of the lungs with positive end expiratory pressure on blood circulation in phthisio-surgical patients]. PMID- 3044193 TI - [Multiple organ failure syndrome]. PMID- 3044195 TI - Use and abuse of blood cultures: program to limit use. AB - Many health care providers order an unnecessary number of blood cultures. We studied the ordering habits in our medical intensive care unit and recommended a protocol for appropriate ordering. Implementation of the protocol resulted in a significant reduction in the number of episodes of suspected sepsis, from 39% of patient discharges during the baseline period to 16% during the study period (p = 0.008). The mean number of blood cultures decreased significantly from 1.2 per patient discharge to 0.3 (p = 0.003). The number of episodes when more than four blood cultures were ordered decreased from 7 to 0, resulting in an annual cost savings estimated at $8025. The net benefit was reversed, however, when the protocol ceased to be actively implemented. We concluded that the appropriate ordering of blood cultures can be effected by establishing a protocol that is actively and continuously implemented. PMID- 3044194 TI - [Rupture of the right external iliac artery by retroperitoneal migration of a total hip prosthesis]. PMID- 3044196 TI - Traumatic aneurysms of cerebral vessels: a case study and review of the literature. AB - Traumatic cerebral aneurysms (TCAs) are rare: few more than 100 cases are recorded in the world literature. TCAs are located predominantly in the supratentorial compartment and are classified as either "true" or "false." A true aneurysm is a dilation resulting from partial disruption of the arterial wall. A false aneurysm results from a full-thickness tear, with the scar from the brain tissue or an organized hematoma acting as the aneurysmal wall. The authors present a case of a false traumatic aneurysm of the pericallosal artery, which was discovered in a young patient fourteen months after a self-inflicted gunshot wound to his head. The aneurysm was an incidental finding on a CT scan performed for the investigation of his late posttraumatic seizures. Its presence was confirmed by angiography. The interval between the traumatic episode and the diagnosis of a TCA usually ranges from a few hours to a few weeks, for most are discovered by angiography performed in the acute or subacute stage of a head injury. The long interval between the injury and the diagnosis in our case is exceptional. A previous CT scan done four months after the injury did not reveal the aneurysm, which strongly suggests a protracted growth of the aneurysmal sac long after the trauma. PMID- 3044198 TI - Issues in the selection of an antiarrhythmic agent. AB - The approach to patients with cardiac arrhythmias involves four steps: diagnosis, determination of whether therapy is needed, choice of appropriate treatment, and evaluation of efficacy of therapy. Factors affecting the decision to treat a patient include the presence and type of symptoms associated with the tachyarrhythmia, as well as the frequency with which the arrhythmia occurs. Antiarrhythmic drugs are usually recommended as first-line therapy although nonpharmacologic approaches are preferable in certain specific situations. There are minimal data for most arrhythmias that aid the physician in the selection of a specific drug for a given patient, and many agents will often be equally effective. Most often drugs are selected that have the least potential to produce harmful side effects to the patient; thus, choice of therapy must be individualized. Newer agents are effective but their antiarrhythmic and side effect profiles will not be understood fully until they have been used widely in diverse patient populations. Regardless of which drug is prescribed, invasive or noninvasive methods should be employed to evaluate therapeutic efficacy. PMID- 3044197 TI - Recurrent giant hypogastric artery aneurysms--a case report. AB - Aneurysms of the hypogastric artery are rare occurrences that are frequently asymptomatic until the time of rupture. When signs and symptoms are present, a pulsatile pelvic mass, frequently detected by rectal or vaginal examination, may produce compression symptoms with urologic, gastrointestinal, and neurologic manifestations. In addition to classical invasive methods of detection such as angiography, newer noninvasive imaging techniques, including ultrasonography, computerized tomography, and magnetic resonance imaging, may be employed to establish the diagnosis. Proximal ligation of the hypogastric artery is the usual method of treatment. A case of bilateral, giant hypogastric artery aneurysms with successful surgical management is reported. PMID- 3044199 TI - Clinical use of sustained-release procainamide in treatment of cardiac arrhythmias. AB - Although procainamide has been available, and used, in the treatment of atrial and ventricular arrhythmias for approximately thirty years, its short half-life has made it less than optimal for long-term arrhythmia management in ambulatory patients. Currently available sustained-release preparations provide the same therapeutic benefit with a more tolerable dosage schedule. The pharmacology of procainamide and its basic electrophysiologic properties, dosage schedules, and toxicity are discussed. Finally, the current indications for its use are reviewed. PMID- 3044200 TI - The role of newer antiarrhythmic drugs in the management of patients with ventricular arrhythmias. AB - The purpose of this review is to discuss the role of mexiletine, tocainide, flecainide, and amiodarone in the treatment of patients with symptomatic ventricular ectopy or sustained ventricular tachycardia (VT) and fibrillation (VF). The pharmacologic properties, electrophysiologic effects, clinical efficacy, adverse reactions, and costs of these new agents are compared with those of procainamide, quinidine, and disopyramide. With the exception of more convenient dosing intervals, these new agents do not afford novel or advantageous pharmacologic properties. The incidences and severity of adverse reactions are similar to those reported with conventional antiarrhythmic drugs. Results of clinical studies demonstrate that these newer drugs are comparable to procainamide, quinidine, and disopyramide in suppressing ventricular ectopy. Accordingly, the choice of an antiarrhythmic agent for patients with symptomatic ventricular ectopy can be based on the side-effect profile. The propensity for developing certain adverse effects can be defined if patients are subgrouped by age and left ventricular function. The adverse effects of amiodarone preclude its use in patients manifesting only ventricular ectopy. Treatment of patients with sustained VT/VF is more complex. Mexiletine and tocainide are generally not effective. The efficacy of flecainide is similar to that of procainamide, quinidine, and disopyramide. The negative inotropic effects of flecainide and disopyramide preclude their use in patients with severe ventricular dysfunction. Amiodarone is the most effective of these drugs for preventing recurrences of sustained VT/VF. Its use should, however, be restricted to patients refractory or intolerant to other antiarrhythmic drugs. PMID- 3044202 TI - [Association between immunity to collagen and the pathology of extracellular matrices]. AB - In a first part, the data of immunogenicity of collagen type II and type IV in physiopathology of extracellular matrices as cartilage and basement membrane, are reviewed. However, in the second part, this conclusion will be examined by basing on the knowledge of the nature of specific sites of these collagens and the variable or non specific humoral immune response towards these collagens in human diseases. PMID- 3044203 TI - [Determination of total serum triiodothyronine in hyperthyroidism: comparison of fluorescence polarization and immunoenzymology]. AB - We evaluated the performances of the Abbott fluorescence polarization assay (FPIA) utilizing the TDx system for human total triiodothyronine (T3) in hyperthyroidism. We compared the results with an immunoenzymometric assay (IEA) (Enzymum Test T3 Boehringer-Mannheim). Greatest attention was focused on the diagnosis of hyperthyroidism because detection of subclinical hyperthyroidism is important. The repeatability of the Abbott fluorescence polarization assay was satisfying (m = 8.07 +/- 0.37 nmol.l-1, CV = 4.59%). The reproducibility was tested with Abbott control sera: m = 4.58 +/- 0.53 nmol.l-1 and CV = 11.5 per cent for level M; m = 7.95 +/- 0.66 nmol.l-1 and CV = 8.23 per cent for level H; m = 2.38 +/- 0.39 nmol.l-1 and CV = 16.5 for level L. The correlation of results of the Abbott assay with those of the Boehringer assay was good for samples from hyperthyroid patients. Values for hyperthyroid and euthyroid subjects were resolved slightly better with the Abbott FPIA than with Boehringer IEA. The Abbott total T3 fluorescence polarization assay may have an additional role to play in monitoring thyroid function in patients under iodine treatment (amiodarone) to eliminate a secondary hyperthyroidism. PMID- 3044201 TI - Mechanisms contributing to arrhythmogenesis during early myocardial ischemia, subsequent reperfusion, and chronic infarction. PMID- 3044204 TI - [The so-called "contact" system of plasma: consequences of the activation of the Hageman factor]. AB - The contact system of plasma includes 4 zymogens, Hageman factor, pre-kallikrein, factor XI and plasminogen, and a cofactor, the high molecular weight kininogen (HMWK). The activation of the contact system leads to the production of bradykinin, blood coagulation, fibrinolysis, complement activation and neutrophil stimulation. Consequently these phenomena generate a lot of pro-inflammatory factors which contribute to the local and systemic inflammatory processes. PMID- 3044205 TI - Fibrin-specific thrombolytic agents. AB - The mammalian fibrinolytic system comprises a proenzyme, plasminogen, which can be converted to the active enzyme plasmin, which will degrade fibrin. Plasminogen activation is mediated by plasminogen activators which are classified as either tissue-type plasminogen activator (t-PA) or urokinase-type plasminogen activator (u-PA). t-PA and single-chain u-PA (scu-PA) induce clot-specific thrombolysis, however via entirely different mechanisms. t-PA is relatively inactive in the absence of fibrin, but fibrin strikingly enhances the activation rate of plasminogen by t-PA. This is explained by an increased activity of fibrin-bound t PA for plasminogen and not by alteration of the catalytic efficiency of the enzyme. scu-PA has a high affinity for plasminogen but, however, does not activate plasminogen in plasma in the absence of a fibrin clot, due to competitive inhibition. Fibrin-specific thrombolysis appears to be due to the fact that fibrin reverses the competitive inhibition, but this does not seem to occur via specific binding of scu-PA to fibrin. The thrombolytic efficacy and fibrin-specificity of natural and recombinant t-PA has been demonstrated in animal models of pulmonary embolism, venous thrombosis and coronary artery thrombosis. In all these studies thrombolysis and relative fibrinogen sparing effect of t-PA was recently confirmed in several multicenter clinical trials in patients with acute myocardial infarction. Specific thrombolysis by scu-PA has also been demonstrated in animal models of pulmonary embolism, venous thrombosis and coronary artery thrombosis. PMID- 3044206 TI - [Treatment of fulminant falciparum malaria with erythrapheresis]. AB - Ten days after his return from Cameroon, a twenty-six year old Frenchman, serving on voluntary service overseas, presented with fulminant falciparum malaria: shock, altered consciousness, haemolytic anaemia, threatening disseminated coagulation (platelets less than 150 X 10(-6).l-1; prothrombin time and Stuart factor less than 50%; fibrinogen less than 1.5 g.l-1). In spite of quinine therapy, parasitaemia increased from 4 to 35% within 24 h. Using an Haemonetics V50, the exchange of one and a half red blood cell masses was carried out with 17 red blood cell packs. Calcium gluconate was used to prevent the hypocalcaemia induced by the anticoagulant solution. The patient's platelets and plasma were completely reinjected. The result was very satisfactory. This kind of exchange, well tolerated clinically and biologically, would seem better than the classical exchange transfusion. When 10% of the red blood cells are infected by Plasmodium falciparum, it is necessary to exchange from one and a half to two blood masses. Lesser exchanges are always associated with important relapses and quinine therapy must be carried on during and after the exchange. Restricting this exchange only to red blood cells enabled the patient to benefit from his own coagulation factors, antibodies and platelets, and consequently to reduce the number of blood donors involved. However, metabolites (especially bilirubin and circulating immune complexes) were not eliminated. Partial plasmapheresis may be associated with erythropheresis using human albumin as plasma substitute. This technique needs to be assessed, in order to optimize immediate efficiency and post-transfusion infectious risk. PMID- 3044207 TI - [Bronchial stenosis of vascular origin in pulmonary arterial hypertension]. AB - A case is reported of bronchial stenosis due to a vascular cause in a patient with chronic obstructive lung disease, cor pulmonale and pulmonary arterial hypertension. This led to right lower lobe atelectasis and acute respiratory failure (pHa 7.24; PaCO2 85 mmHg; PaO2 44 mmHg) with important right-to-left shunting. This diagnosis was only suggested on day 7 by fibreoptic bronchoscopy and confirmed a week later by tomography and digital angiography. Nifedipine, used to reduce the pulmonary arterial hypertension, increased the cardiac index (31.min-1.m-2 to 3.3.1.min-1.m-2) and oxygen transport (488 ml.min-1.m-2 to 554 ml.min-1.m-2), despite increasing the shunt effect (Qs/QT: 26% to 31%). This and the antiinflammatory action of methylprednisolone were probably responsible for the favourable outcome. PMID- 3044209 TI - Intragastric balloons: an unfulfilled promise. PMID- 3044208 TI - Nonsteroidal anti-inflammatory drugs: gastropathy, deaths, and medical practice. PMID- 3044210 TI - Hospital-associated infection from leeches. PMID- 3044211 TI - The medicinal leech. A page from the annelids of internal medicine. AB - Leeches have been used in health care since ancient times by physician and layman alike. As just one of several methods of bloodletting, the leech became the focus of a science that included such subjects as indications, modes of attachment, complications, and relative contraindications. The popularity of leeching has varied immensely over the years. In the 19th century, this annelid saw its numbers decimated because of protean medicinal indications. The leech lost its hold on the practice of medicine in the early 20th century. Recently, the use of leeches has resurged in both the lay and the scientific communities. PMID- 3044212 TI - Adults with cyanotic congenital heart disease: hematologic management. AB - Hematologic management of adults with cyanotic congenital heart disease has received little recent attention. The lack of practical therapeutic guidelines prompted us to consolidate our observations on 124 cyanotic adults for general physicians, cardiologists, and hematologists who care for these patients. Specific attention focused on regulation of erythrocyte mass and concepts of compensated and decompensated erythrocytosis, symptoms of deficient tissue oxygen transport, hyperviscosity and iron deficiency, the potential relation between elevated hematocrit levels and brain injury, hemostasis, urate metabolism, and renal function. Cerebral infarction was not seen in any patient. Phlebotomy is best reserved for treatment of symptomatic hyperviscosity. Iron therapy is indicated for symptomatic iron deficient erythropoiesis. Abnormal hemostatic mechanisms are the rule. Antithrombotic medications have little or no role in treatment. Hyperuricemia is the result of abnormal renal uric acid excretion not urate overproduction, and serves as a marker of abnormal renal function. Drugs that promote urate excretion are the preferred maintenance treatment in symptomatic hyperuricemic patients. PMID- 3044213 TI - The care of elderly patients with cardiovascular disease. AB - Cardiovascular disease is a major clinical problem in the elderly, with coronary heart disease the most frequent cause of death and with hypertension present in as many as 50% of these patients. The cardiovascular manifestations of aging must be differentiated from those due to disease. There are clinical manifestations and responses to therapy in the elderly that differ from those in younger patients. The extent of diagnostic and therapeutic procedures undertaken should be based on the patient's physiologic age, the presence and severity of concomitant diseases, mental status and cognitive ability, and the patient's expectations from medical care. Preventive approaches are also warranted. PMID- 3044214 TI - Toward a romantic science: the work of Oliver Sacks. PMID- 3044216 TI - Privatization of health care: a U.S. perspective. PMID- 3044217 TI - Protection of human subjects of biomedical research in the United States. A contrast with recent experience in the United Kingdom. AB - In the U.S., the development of extensive regulations for the protection of human subjects of research began in the 1960s and continued through the 1970s. The substance of these regulations reflects the American social and political climate of the time. There is a focus on rights--e.g., to be left alone, to be self determining--reflected in elaborate requirements to assure the validity and documentation of informed consent. There is also a focus on systems of disinterested review and monitoring procedures to assure uniform adherence to the requirements of the regulations. To the extent that the U.S. has developed extensive regulations in this field, it may be viewed as more advanced than the U.K. And yet, it is apparent that there remain on both sides of the Atlantic very difficult and similar problems regarding the definition of responsible research. Such problems are illustrated by consideration of current controversies about the ethical justification of RCTs. There are some features of the U.S. regulatory system that I can commend to the attention of other nations as they develop policies for the protection of human research subjects. For example, a uniform requirement for informed consent and committee review appears to be responsive to some problems currently encountered in the conduct of RCTs in the U.K. A note of caution is in order, however. Some features of our regulatory policy and practices are excessively inflexible, wasteful of human resources, and occasionally counterproductive. PMID- 3044218 TI - Medical research and the human subject. Problems of consent and control in the UK experience. PMID- 3044219 TI - A tale of two cultures; do we differ? PMID- 3044220 TI - Biomedical ethics: an Anglo-American dialogue. A personal view of future trends. PMID- 3044221 TI - Two branches from one stem. PMID- 3044222 TI - Assisted human reproduction and embryonic surgery: the ethical issues. PMID- 3044223 TI - Medical ethics in its American context. An historical survey. AB - Until the 1950's, moral aspects of clinical practice were handled in the USA within the medical profession. Over the last 30 years, these issues have become subjects for public debate, and have changed the public perception of medicine, in four steps. In the 1950's, moral theologians questioned the implications of medical technology at the edges of life. In the late '60s and '70s, these theologians were joined by political activists, whose zeal provoked a counter reaction from physicians. In the late '70s and early '80s, the debate became largely theoretical; but in the late '80s it is once again "clinical", though respecting the rights of patients, their families, and other nonphysicians to participate in the relevant moral decisions. In part, these four steps reflect the special feature of American social history in the last 30 years; but in part they also had counterparts in Britain and elsewhere. Either way, the monopoly control over the ethics of medical practice exercised by doctors before the 1950s is unlikely to return. PMID- 3044224 TI - A computer-assisted instruction course to teach visual-field interpretation. AB - A computer-assisted instruction course for visual-field interpretation is described. A multiple-choice, question-and-answer drill was constructed using the VAX/VMS minicomputer and commercially available educational software. Medical and paramedical personnel can use this course to further their knowledge of visual field description, the location of lesions which lead to visual-field defects, and the underlying processes that cause some of these defects. The advantages of using a particular computer terminal because of its graphic capabilities and screen layout are described. Other uses of computer-assisted instruction in ophthalmology are discussed and encouraged. PMID- 3044226 TI - Antiglaucoma therapy during pregnancy--Part II. PMID- 3044225 TI - Keratoplasty in children. AB - One hundred sixty-two eyes of children less than 14 years of age with corneal disease underwent keratoplasty over a period of eight years. The cases were divided into two groups and different subgroups, and the results were compared. The results of the recent group of cases after introduction of microsurgery were definitely better. However, being a referral center, the cases came to us at a late stage with complications and required larger grafts. Interestingly, graft transparency has a direct correlation with the size of the graft and type of corneal disease. Lastly, there was a marked reduction in frequency of therapeutic keratoplasty later, a result of the availability of various drugs. PMID- 3044227 TI - Dynamic amaurosis fugax secondary to compression of vertebral artery. AB - Transient unilateral loss of vision can be a manifestation of ipsilateral carotid disease; whereas transient visual impairment affecting both eyes simultaneously suggests vertebral basilar-artery insufficiency. We present such a case where the symptoms were related to head position. Congenital anomalies of the vertebral arteries were found on four-vessel cervical angiography. PMID- 3044228 TI - A-scan ultrasonography in the diagnosis of orbital dermoid cysts. AB - Eleven patients with anterior and deep orbital masses had A-scan ultrasonography. The ultrasonograms were all similar and demonstrated a dermoid-like pattern of low internal reflectivity and a high-rising, double-peaked posterior surface spike. Although the postoperative pathologic findings in seven cases confirmed the clinical diagnosis of dermoid cyst, other lesions, such as epidermoid cyst, sebaceous cyst, Graves's myopathy, and Ewing's sarcoma were found to show a similar ultrasonographic pattern. This stresses the importance of computed tomography in the diagnosis of orbital tumors. PMID- 3044229 TI - [Did Napoleon suffer from sleep apnea syndrome?]. AB - Napoleon would sleep very little. He frequently woke up during night and worked. Brief sleeping time in day repaired his fatigue. He had also a short and thick neck. In the last fourth of his life he progressively suffered from obesity, daily involuntary sleepiness and his intellectual capabilities undoubtedly decreased. Our experience of 48 cases of sleep apnea syndrome diagnosed by mean of polysomnography allow no to think that Napoleon suffered from this disease. Historical consequences of this pathology is discussed. PMID- 3044230 TI - Functional disorder of eustachian tube in experimental otitis media with effusion following inoculation of bacterial endotoxin. AB - A 10-micrograms/mL solution of lipopolysaccharide derived from Klebsiella pneumoniae was inoculated into the middle ears of guinea pigs. The animals were killed painlessly on the first, third, or seventh day after inoculation, and the mucosal samples from the bony portion of the eustachian tube were examined for ciliary activity and epithelial morphology. On the first and third days, when middle ear effusions were present, deterioration of ciliary activity and morphologic changes in the mucociliary system were observed. On the seventh day, when middle ear effusions were absent, the ciliary activity had recovered to normal. Our data show that endotoxin extracted from K pneumoniae can produce otitis media with effusion and that dysfunction of cilia caused by endotoxin is a factor responsible for the manifestation of otitis media. PMID- 3044231 TI - [Neurocutaneous Degos disease]. PMID- 3044232 TI - [Contraception in the woman with lupus erythematosus]. PMID- 3044233 TI - [Gingival hypertrophy]. PMID- 3044234 TI - [Cyclosporin in dermatology]. PMID- 3044235 TI - Cerebral ischaemia and surgical practice. AB - This review article examines the principles underlying the control of cerebral blood flow, the consequences of brain ischaemia and subsequent reperfusion and discusses the impact of brain ischaemia in a number of clinical situations of surgical importance. PMID- 3044237 TI - Bacterial colonisation of leg ulcers and its effect on the success rate of skin grafting. AB - There is little information on the immediate and long-term results of skin grafting to chronic lower limb ulcers. Our experience in their management had led us to analyse, retrospectively, the results of split thickness skin grafts applied to lower limb ulcers in 88 consecutive patients. Graft take has been related to bacterial growth from ulcer swabs taken on admission, preoperatively and postoperatively. Follow-up was for a median of 18 months. Initial graft take varied from 20% to 100% (median 85%). Bacterial flora grown from the ulcer swabs varied with the duration of the ulcer and the treatment. Analysis by bacterial type has shown that Staphylococcus aurcus and Pseudomonas significantly reduced skin graft healing. Overall, 90% of these ulcers had healed with a median of 6 weeks' in-patient treatment. Examination of the swab results from the 8 ulcers that were slow to heal postoperatively and the 8 ulcers that recurred 6 days to 8 months after discharge from hospital revealed that 15 out of 16 (94%) grew S. aurcus; none had Pseudomonas isolated from them. After eighteen months 8% of these ulcers remain active. Aetiology appears important as this figure is 6% for limbs affected by venous disease only, 13.3% for limbs with arterial disease only and 13.6% for limbs with both venous and arterial disease. PMID- 3044236 TI - The effect of dopamine on renal function during aortic cross clamping. AB - Eighteen male patients undergoing elective surgical reconstruction of the abdominal aorta were divided into two groups. Patients in Group I (nine) were given dopamine intravenously, in a dose of 2 micrograms/kg/min, during the first half of the period of cross-clamping, whilst those in Group II received dopamine during the second half. Each patient acted as his own control and for each, three periods were examined, namely: pre-clamp, clamping with dopamine and clamping without dopamine. Dopamine infusion during aortic clamping caused a significant rise in sodium output (P less than 0.01), potassium output (P less than 0.05), creatinine clearance (P less than 0.05) and urine output (P less than 0.05). We conclude that dopamine infusion during aortic clamping helps to protect the kidney from any deleterious effect of clamping. PMID- 3044238 TI - Comparative evaluation of general, epidural and spinal anaesthesia for extracorporeal shockwave lithotripsy. AB - The results of a prospective randomised evaluation of general anaesthesia (GA), epidural anaesthesia (EA) and spinal anaesthesia (SA) for extracorporeal shockwave lithotripsy are presented. GA provided speed and reliability but resulted in a high incidence of postoperative nausea, vomiting and sore throat. Both regional techniques conferred the advantages of an awake, cooperative patient, but EA required a longer preparation time than SA and more supplementary treatment with fentanyl or midazolam. A major drawback associated with the use of SA was a 42% incidence of postspinal headache. All three techniques were associated with hypotension on placement in the hoisl; bath immersion resulted in significant rises in blood pressure in the EA and SA groups and a more variable (overall non-significant) response in the GA group. PMID- 3044240 TI - Digital subtraction cholangiography: a new technique for visualising the common bile duct during cholecystectomy. AB - Operative cholangiography is for most surgeons a routine part of every cholecystectomy. Computerised digital subtraction angiography was adapted for operative cholangiography using a portable machine. After cannulation of the cystic duct the background image was subtracted before injecting contrast. Only the contrast within the bile duct appears on the monitor and resolution is high. A permanent record was made on 10 X 10 cm spot films. Eighteen pre-exploratory cholangiograms were performed using this method. In 12 no stones were demonstrated on digital subtraction cholangiography (DSC), nor were there clinical indications of common bile duct stones. These patients underwent cholecystectomy only. Stones were demonstrated on DSC in 3 patients and all had stones at exploration of the common bile duct (CBD). Three patients had no stones demonstrated on DSC but were explored on clinical grounds. No stones were found. Postoperative T-tube cholangiograms confirmed the absence of stones in 5 patients. A retained stone was present in one patient who had not had a postexploratory examination at operation and was not related to the use of this cholangiographic technique. DSC combines the benefits of image intensification and still radiography and has been accurate in both predicting and excluding common bile duct stones. PMID- 3044239 TI - Splanchnic ischaemia and multiple organ failure in the critically ill. AB - The development of a safe and simple tonometric method of monitoring the adequacy of gastrointestinal mucosal perfusion has provided a new perspective of splanchnic ischaemia in the critically ill. It would appear that splanchnic hypoxia, identified by the presence of intramucosal acidosis in the gut, may be one of the most consistent and earliest indications of impaired tissue perfusion in the critically ill and be causally related to the development of sepsis and multiple organ failure. PMID- 3044241 TI - Early experience with B mode ultrasound mapping of the long saphenous vein prior to femorodistal bypass. AB - B mode ultrasound was used to assess and map the long saphenous vein in 20 limbs prior to femorodistal bypass. The assessment was compared with operative findings. Eighteen of 19 adequate veins and 8 of 9 anatomical abnormalities or major divisions were correctly identified. B mode ultrasound allows accurate marking of the vein, facilitating dissection, alerts the surgeon to possible difficulties and is an ideal non-invasive technique for preoperative assessment of the long saphenous vein. PMID- 3044243 TI - [Multiple proximal coronaro-pulmonary fistulae. Review of the literature apropos of a new case]. AB - The authors report a new case of multiple proximal coronaro-pulmonary fistula between right coronary arteries, anterior interventricular artery and the trunk of the pulmonary artery, in a 64 year-old female patient with chest pain and a continuous murmur located in the third left intercostal space. The coronary steal is demonstrated by a myocardial scintigraphy during stress with return to normal after surgical ligation. A review of the literature enabled to find 33 cases of this major congenital anomaly of the coronary arteries, defined as an abnormal communication between at least two main coronary vessels and the trunk of the pulmonary artery. This results in a left-right shunt, usually minor without any repercussions on the right cavities and pulmonary pressures. The entire clinical, electrocardiographic, radiological, sonographic, scintigraphic, haemodynamic and angiographic picture is reported for these 33 cases. A physiopathological discussion is proposed. The course of this disease is usually favorable (only one case of myocardial infarction was published, without cardiac failure. Osler's endocarditis or sudden death); this seems to authorize simple monitoring as a logical therapeutic approach except when a myocardial ischemia secondary to coronary steal is demonstrated, imposing a surgical correction. PMID- 3044242 TI - Bupivacaine squirting. PMID- 3044244 TI - [Cerebral embolism of cardiac origin]. AB - Embolisms of cardiac origin represent approximately 15 p. cent of the causes of cerebral vascular accidents. This article considers the difficult diagnostic and therapeutic problems raised by cerebral embolisms. First: arguments permitting to suspect this mechanism in the absence of an obvious cardiac cause or in the presence of a high incidence heart disease but low embolic potential. The main etiologies are then reviewed emphasizing the evolution of their respective frequencies, then a program of additional investigative examinations is suggested. Finally the indication of anticoagulants is discussed according to neurological and cardiologic criteria. PMID- 3044245 TI - [Invasive and non-invasive methods for the diagnosis of tachycardia]. AB - Surface ECG permits, in most cases, to determine the ventricular or supraventricular origin of a tachycardia with wide complexes, to individualize VTs with aspects of Left Buble Branch and left axis, to visualize epsilon waves in some chronic VTs. The late potentials detected by the sum-mean method seem to be well correlated with the occurrence of ventricular tachycardia, especially after myocardial infarction. The area of origin of a VT may be determined quite precisely by cartography and topographic stimulation. The predictive value of the provocation methods (stress test, transesophageal stimulation, endocardiac stimulation) is discussed. Some arrhythmias may be discovered on ultrasonic cardiography and the exact mechanism of some reciprocal permanent junctional tachycardias is well demonstrated with endocardiac stimulation and fulguration. Nuclear magnetic resonance offers a new possibility of etiological diagnosis in ventricular tachycardias with left delay. PMID- 3044247 TI - [Acquired systemo-pulmonary fistula after pleurectomy responsible for a continuous thoracic murmur]. AB - Systemo-pulmonary fistulae are rare. The case of a 27 year-old man, hospitalized for exploration of a continuous thoracic murmur, is reported. A right pleurectomy had been performed 2 years previously because of a recurrent spontaneous pneumothorax, and no murmur was present at that time. Angiography showed a systemo-pulmonary fistula with the right internal mammary artery and branches of the right axillary artery as afferent vessels, and the right pulmonary arteries and veins as efferent vessels. Blood gases measurements demonstrated a left-right shunt. The acquired nature of the fistula was suspected because of the history of right pleurectomy and the acquired nature of the murmur. There was no indication for surgery because of the complexity of the fistula and the absence of symptoms. PMID- 3044246 TI - [Fulguration of foci of atrial tachycardia in the adult]. AB - Fulguration of the site generator of arrhythmia was attempted in three 31, 33 and 55 year-old patients presenting ectopic atrial tachycardias resistant to various antiarrhythmic medications. Episodes of atrial fibrillation and flutter were also documented in two of them. Two patients had a surgically corrected congenital cardiopathy:interatrial communication or pulmonary stenosis. Mapping of the endocardial emergence point was carried out with electrodes placed 1 cm apart, demonstrating the high atrial origin of the tachycardia, near the right atrium. The auriculogram preceded the earliest possible ectopic P wave by 20 to 70 ms; its multiphasic and prolonged morphology, suggested a local intra-atrial conduction disorder in the three cases. Cathodic shocks were delivered at this site without complications with cumulative energies of 720 J, 480 J and 320 J, respectively. The fulguration was ineffective and revealed other arrhythmic sites in two patients. Only patient n1 has been asymptomatic since 24 months under a treatment with Sotalol 160 mg which had been previously ineffective. PMID- 3044248 TI - [European pioneers of modern cardiology]. PMID- 3044249 TI - [Cardiac Doppler ultrasonography in the diagnosis and surveillance of chronic aortic insufficiency]. AB - Chronic aortic insufficiency is, since a few years, a topic of renewed interest because of the development of non-invasive monitoring methods: sonocardiography, cardiac Doppler. The problem raised is that of deciding the most opportune time for surgery in asymptomatic chronic AI, the course of which may quietly evolve towards irreversible myocardial lesions. The severity of AI may be determined by numerous sonocardiographic and Doppler criteria. In sonocardiography, three types of parameters have especially been studied: ventricular expansion (telediastolic, telesystolic diameter), the systolic function (percentage of diameter decrease), volume/mass ratio (R/h). To be valid, these measurements are a precious guide for patient's monitoring and therapeutic approach. The more recent Doppler criteria must be validated on large series and evaluated according to the context or a pathological association; they all have limitations and cannot be interpreted separately. PMID- 3044250 TI - Minerals, trace elements and cardiovascular disease. An overview. PMID- 3044251 TI - More exercise for the diabetics? Physical activity and prevention of ischaemic heart disease. AB - Epidemiologic studies suggest that a high level of physical activity would protect against coronary heart disease. There is accumulating evidence that habitual physical training may reduce some risk factors of atherosclerosis in both non-diabetic and in diabetic subjects. In this article the relationship of exercise to cardiovascular risk factors in diabetic patients is reviewed. PMID- 3044252 TI - Immunology--directions for the future. AB - This brief introduction highlights how far immunology has moved as an independent discipline. From its birth as a branch of medical microbiology, it now pervades virtually the whole of medicine and many of its subspecialties. The steps along this pathway are summarised, and the main challenges for the future outlined. PMID- 3044253 TI - Colony stimulating factors and hemopoiesis. AB - Work in the past twenty years has led to the discovery of multiple families of hemopoietic growth factors that regulate the production and functional activity of the various subsets of blood cells. For granulocyte-monocyte populations, four such regulators (the colony stimulating factors GM-CSF, G-CSF, M-CSF and Multi CSF) have been shown to interact to regulate these populations. Each has been purified, cDNA's for each have been cloned and mass-produced recombinant CSF's are becoming available for clinical use. Initial trials indicate that the CSF's will be valuable agents in stimulating hemopoiesis either following chemotherapy or marrow transplants or in stimulating resistance to life-threatening infections. CSF's play an important role in the development of myeloid leukemia but curiously can suppress myeloid leukemic populations because of their differentiation-inducing actions. However, it is still unclear whether the CSF's will prove of value in the management of particular myeloid leukemias. PMID- 3044255 TI - Immunoparasitology: its contribution to development of new parasite vaccines. AB - Immunoparasitology--the study of the immunology of host-parasite relationships- can post some notable research successes over the past decade. Progress towards prophylactic molecularly-defined vaccines against human parasitic diseases such as falciparum malaria, schistosomiasis mansoni and cutaneous leishmaniasis, as well as economically-important veterinary parasites, has been good. However, new vaccines are not coming as easily as might be hoped mainly because of several deficiencies in knowledge on the immunology of host-parasite relationships and the unknown relevance of well-characterized model systems to real-life parasitic diseases. In some models, the immunology of resistance and the immunology of disease are understood in broad outline. The availability of isolated antigens and their epitopes has improved quantitation of host immune responses to various life cycle stages of parasites and enabled vaccination efficacy or diagnostic potential to be assessed. One of several major challenges facing the immunoparasitologist interested in vaccine development is overcoming genetically based unresponsiveness to "oligoepitope", defined-antigen vaccines particularly at the level of helper TH and cytotoxic (Tc) T cells. PMID- 3044256 TI - Efficacy of hepatitis B immunisation. AB - Hepatitis B virus (HBV) infection is endemic to Singapore, the carrier rate being highest among the Chinese. It is the cause of significant morbidity and mortality. The prevention of this condition is therefore an important objective. Plasma derived vaccines have been shown to be effective in the prevention of perinatal as well as horizontal spread of HBV. However, the high cost of production together with deep-seated, though unfounded fears of the possibility of transmission of other viruses has limited its use. The advent of recombinant yeast vaccine is therefore an important event since the unfounded fears can now be laid to rest, and the cost of immunisation should eventually decrease as production increases. The efficacy of recombinant yeast vaccine has been found to be comparable to that of plasma derived vaccine. PMID- 3044254 TI - Modern approaches to vaccine development with special reference to the needs of developing countries. AB - Vaccination has proved to be one of the most effective public health measures to control infectious diseases. The eradication of smallpox by world-wide vaccination represents one of mankind's greatest achievements. Despite the availability of vaccines to control many diseases, they are generally under-used in many developing and some developed countries. However, there are many diseases for which current vaccines are inadequate or vaccines cannot be prepared using conventional approaches. This article describes the new approaches which are now available and are being used extensively to develop new vaccines against viral, bacterial and parasitic diseases. Success has already been achieved in a few cases and the prospect for others is encouraging. In addition, progress is being made to develop vaccines to control human fertility as this development is seen to complement the control of infectious diseases. PMID- 3044257 TI - Clinical evaluation of a yeast recombinant hepatitis B vaccine in healthy hospital staff in Singapore. AB - A yeast recombinant hepatitis B vaccine (subtype adw), derived from purification of HBsAg particles, expressed in the yeast Saccharomyces cerevisiae which contained the gene for HBsAg, was evaluated in 31 healthy adult hospital staff members in Singapore. Each subject received a 10 mcg dose of vaccine intramuscularly at 0, 1 and 6 months. One month after the first two injections of vaccine the seroconversion rate (defined as greater than or equal to 2.7 mIU/ml) was 90%. Two months after the third injection 100% of participants had anti-HBs levels higher than 2.7 mIU/ml and 97% had titers of anti-HBs greater than 10 mIU/ml. The geometric mean titer (GMT) of anti-HBs levels at 2, 3, 6, 8 and 12 months were 21.9, 38.6, 57.6, 1253.4 and 354.1 mIU/ml, respectively. All clinical complaints were mild and transient. They consisted of mild soreness at the injection site, transient fever and headache. There was no correlation between the presence of antibodies to S. cerevisiae with any allergic manifestations. The vaccine was safe and immunogenic for staff exposed to an infection risk and should now be widely used in the extension of hepatitis B immunization programs. PMID- 3044258 TI - Autoantibodies: the search for origins, target autoantigens and pathogenicity. AB - The origins of autoantibodies remain largely unknown. Burnet suggested that self reactive clones of B lymphocytes are deleted during ontogeny and that 'forbidden' clones of autoantibody-producing B lymphocytes arise as a consequence of somatic mutation of the variable regions of immunoglobulin genes. The alternative view, which is gaining increasing support, is that autoreactive B cells are present in all normal individuals but are held in check by control mechanisms, e.g. by the idiotypic network of Jerne. Autoantibodies segregate with clusters of autoimmune diseases and are useful diagnostic markers of these diseases. The autoantigens targeted by these autoantibodies are largely unknown. The precise molecular characteristics of these autoantigens are currently under active investigation by immunochemical and recombinant DNA methods. Precise definition of these autoepitopes should lead to sensitive immunoassays and studies directed at determining the pathogenicity of the autoantigens. The new knowledge gained in this way may lead to successful restoration of tolerance to self antigens. PMID- 3044259 TI - Autoantibodies in systemic lupus erythematosus. AB - Systemic lupus erythematosus (SLE) is a multisystem autoimmune disease in which the predominant autoantibodies are antinuclear antibodies (ANA) reactive with DNA and histones, and antibodies reactive with the non-histone extractable nuclear antigens (ENA), Sm and Ro. Racial differences demonstrable in predisposition to SLE are also evident in the prevalence of autoantibodies, the frequency of anti Sm and anti-Ro being 2-4 times higher in Asians with SLE than in Caucasians with SLE. Autoantibodies have also played a role in the classification of lupus and the recognition of multisystem autoimmune diseases that fail to meet the classical criteria for the identification of patients with SLE but appear to be variants of lupus. Anti-Ro is a diagnostic marker for subacute cutaneous lupus, anti-(U1)RNP a marker for mixed connective tissue disease, antibodies to phospholipids a marker for the syndrome comprising stroke, fetal wastage, thromboembolism and thrombocytopenia and antibodies to histones a marker for lupus induced by the drugs hydralazine and procainamide. There is still no unaminity on whether these antibodies play an integral role in the disease process or whether they are "epiphenomena". The challenge for research in the 1980s is the understanding of the relationship of these antibodies to the pathogenesis of SLE. PMID- 3044260 TI - Monoclonal antibodies and the lymphoid malignancies. AB - A comprehensive classification of the lymphoid malignancies with correlation between histology, clinical disease and with prognostic value has proved difficult to formulate, partly because of the heterogeneous nature of these neoplasms, inadequate definition of non neoplastic cells in the admixture of neoplastic cells and also as a result of the paucity of special techniques to supplement light microscopy in the study. The introduction of monoclonal antibodies against human leucocyte antigens for diagnosis in the lymphoid malignancies using the immunofluorescence or immunoperoxidase technique has provided a basic working immunological formulation and greatly advanced understanding of the biology of these diseases and the maturational staging of normal B and T lymphocytes. This review briefly looks into recent advances and presents a simplified approach to understanding this rapidly advancing field especially for clinicians. PMID- 3044261 TI - Immunologic mechanism in contact allergy--a review. AB - The last decade saw rapid advances in the understanding of the immunology of contact allergy, a cutaneous delayed hypersensitivity reaction. Of great importance was the recognition of the role of epidermal Langerhans cells as antigen-presenting cells to T lymphocytes. The necessity of an intact lymphatic system to mount contact allergy was demonstrated. The identification of lymphokines, mediators and modulators of inflammatory response in contact allergy, enable us to understand the histological and clinical manifestation of allergic contact dermatitis. Based on the discovery of the immunologic pathways of contact allergy successful attempts have been achieved to prevent and treat allergic contact dermatitis by blocking or modifying its pathways. This paper reviews recent development in the immunologic mechanism of contact allergy. PMID- 3044262 TI - Gamete intrafallopian transfer (GIFT): an alternate method of conception in couples with unexplained infertility, endometriosis and oligo-asthenospermia. AB - 138 treatment cycles of GIFT were carried out in couples with mainly unexplained infertility, endometriosis and male factor. The overall pregnancy rate was 24.6%, abortion 29.2%, multiple pregnancy 20.6% and ectopic pregnancy 0%. Four oocytes seem to be the optimal number of oocytes to be transferred. Patients with male factor infertility fared the worse in comparison with the other groups. Successful in-vitro fertilization of excess eggs did not correlate with a positive outcome of the GIFT procedure. PMID- 3044263 TI - Immunology in Malaysia--a review. AB - Immunology is a discipline that traverses all branches of clinical medicine. Thus since about ten years ago major hospitals in Malaysia established routine clinical immunology services particularly in the diagnosis of autoimmune/connective tissue disorders. More recently these laboratories have ventured into basic research in Dengue Haemorrhagic Fever, Leukaemia Immunology, Nasopharyngeal Cancer and Leprosy. The rationale for these projects together with early results from them are discussed. PMID- 3044264 TI - Common variable immunodeficiency--a case report and review. AB - Common Variable Immunodeficiency is a rare primary immunodeficiency presenting usually in young adults with repeated sinopulmonary infections as a result of profound hypogammaglobulinemia. A clinical report of the first documented patient in this region is presented along with a brief review of the recent advances made especially in understanding pathogenesis and management of patients with this disorder. PMID- 3044265 TI - Hepatitis B virus (HBV)--the challenges ahead. PMID- 3044266 TI - Assessment for rehabilitation after stroke. AB - There is some evidence that the rehabilitation of patients who have suffered a stroke may be more successful if conducted in a specialist unit, especially if started soon after the stroke. Departments of Geriatric Medicine are often unable to accept all invitations to take over the rehabilitation of elderly patients who have suffered a stroke. As specialist rehabilitation resources are scarce, it is necessary to be able to distinguish the patients whose outcome will be improved by specialist rehabilitation from those whose recovery (whether good or bad) will not be significantly influenced. Patients with an Edinburgh prognostic score of 5.6 four weeks after the stroke are very likely always to require total nursing care even after specialist rehabilitation. A Guy's Hospital prognostic score of 10 or more seems to indicate that functional independence will be recovered even without specialist rehabilitation. Patients with an Edinburgh prognostic score of 3-4 within the first 4 weeks after the stroke should not be denied specialist rehabilitation; there is evidence, admittedly weak, that it is for them that the benefits may be greatest. A major study is now required to validate the Edinburgh and Guy's prognostic scores prospectively and to test whether it is valid to use them for the triage of elderly patients for specialist rehabilitation. In addition, it is possible that a specialist unit would be better able to help patients with specific 'barriers' to rehabilitation. Adequate tools for the identification and objective assessment of these barriers are essential if this possibility is to be tested. PMID- 3044267 TI - Mitral valve prolapse--a review. AB - Mitral valve prolapse is one of the more common cardiac conditions in our community. The prevalence is difficult to assess due to differences in diagnostic criteria and population selection. The prevalence in the community has been estimated to be from 5 to 15%. In females there is a decline in prevalence with increasing age which is not observed in males. 30% of subjects with mitral valve prolapse have relatives with the same condition. Strahan et al suggested that mitral valve prolapse was inherited as an autosomal dominant trend with its manifestations modified by age and other factors. PMID- 3044268 TI - Biomarkers of xenobiotic exposures. AB - Direct measurement of xenobiotic (foreign) chemicals is not always feasible as an exposure assessment,--owing to rapid metabolism, sequestration into fatty tissues, or lack of suitable assay methods. Furthermore, suspect exposures often involve complex mixtures of organics. In these circumstances, indirect biomarkers of exposure can be most helpful. This paper reviews four urinary parameters that hold promise as biomarkers of exposure in occupational and environmental settings: glucaric acid (end-product of the glucuronidation pathway), thioethers (end-product of glutathione reaction with electrophilic or alkylating agents), porphyrin pattern (altered with disruption in heme biosynthesis), and the Ames mutagenicity test. PMID- 3044269 TI - Involvement of hormones in the swift increase in alcohol metabolism. AB - The purpose of this study was to determine the time course of changes in blood levels of various hormones in C57BL/6J mice during exposure to ethanol vapor. Groups of adult male mice were given 2.0 g per kg ethanol intraperitoneally or as continuous vapor for four hours and rates of ethanol elimination were measured. In parallel, blood samples were collected at timed intervals over 5.5 hours during and following exposure to ethanol. Blood levels of epinephrine, norepinephrine, corticosterone, and glucagon were elevated two- to four-fold during ethanol treatment and declined to basal values within one hour following termination of treatment. Elevated blood levels of epinephrine, norepinephrine and corticosterone were highly correlated with higher rates of ethanol elimination (r = 0.80, 0.78, and 0.72, respectively). In contrast, thyroxine and insulin levels were not affected by ethanol. These findings are consistent with the idea that acute administration of ethanol causes the release of glycogenolytic hormones which in turn increase rates of ethanol metabolism. PMID- 3044270 TI - [Medullary cancer of the thyroid: morphological and immunohistochemical definitions. The pathologist's role in 1987]. AB - The morphological and secretories aspects of medullary thyroid carcinomas are reviewed. Mixed follicular and medullary carcinomas are discussed. Criteria for discrimination between sporadic and hereditary forms are mentioned with special reference on C cell hyperplasia. After a brief report on what is known about normal human and animal C cell, a short histogenetic discussion is debated. PMID- 3044271 TI - [Diagnostic circumstances, therapeutic measures and long-term surveillance of medullary cancers of the thyroid]. AB - Diagnosis of medullary thyroid carcinoma is often delayed as a consequence of its rarity and its recent discovery. Fine needle aspiration biopsies of all the cold thyroid nodules and plasma calcitonin (CT) measurement in case of suggestive clinical features allow an accurate pre-operative diagnosis and the best therapeutic conditions. A cervico-mediastinal check-up, a screening for a Multiple Endocrine Neoplasia type 2a or 2b, the removal of all tumoral cervico thoracic tissue by total thyroidectomy, a careful node excision and the pathological examination by a skillful pathologist using immunohistochemistry are the main therapeutic methods. Additional treatment such as external radiotherapy, chemotherapy and radiopharmaceutics are indicated when surgery is incomplete, when the tumor is inoperable and when there are extensive distant metastases. Basal and pentagastrin stimulated CT is the best tool in post surgical follow-up. Disappearance may indicate a long remission, even a cure. Persistence of an abnormal level necessitates the search for secreting tumoral site with non invasive methods such as ultrasonography, computed tomography and scintigraphy, avoiding blind aggressive treatments. In case of recurrence or distant metastases, surgical excision has to be considered first. PMID- 3044272 TI - Birth weights in three Norwegian cities, 1860-1984. Secular trends and influencing factors. AB - Data from birth records from three maternity hospitals in Norway have been used to study the trend in birth weight in this country from 1860-1984. The investigation is based on a sample of 200-300 records taken at random from 2 to 5 years around every 10th year, from each of the three maternity hospitals- amounting to a total sample of 9152 women. Besides describing the trend in birth weight, I have analysed the different factors influencing birth weight--using multivariate linear regression methods. The results show that the mean birth weight has changed remarkably little throughout this period of 120 years. The total increase has only been just below 200 g. From Montreal, Ward and Ward (1984) have reported a decrease in mean birth weight of about 430 g during the second half of the last century. The Norwegian data indicates a simultaneous fall in mean birth weight of about 70 g. This fall is, however, found mainly among unmarried women. Thus, the great decline in birth weight which is found in Montreal, is not apparent in our material from Norway, despite the fact that Oslo was industrialized at approximately the same time as Montreal. Of the various independent variables used in the multiple regression analyses, only the following variables appear to be of any importance: the year when the birth took place, the mother's menarcheal age, her marital status, the sex of the child and the parity number. In this material, I have found a linearly increasing birth weight from birth number 2 up to at least parity 8, and a larger increase from parity 1 to 2. The birth weight shows a significant increase in the case of married women as opposed to unmarried. This effect is most marked before 1900, indicating that the social conditions gradually became less unequal after the turn of the century. The birth weights have a decreasing tendency with higher ages of menarche. This tendency is constant throughout the whole period of time. Boys appear to be about 112 g heavier than girls, this difference being largely the same throughout the investigated period. PMID- 3044273 TI - [Comparison of methods for evaluating the nephrotoxicity of cisplatin]. AB - The BUN, serum creatinine, creatinine clearance and the urinary excretion of leucine aminopeptidase (LAP), alkaline phosphatase (ALP), beta 2-microglobulin (beta-m), and N-acetyl-beta-glucosaminidase (NAG), were measured in 21 gynecological cancer patients treated with CAPF (CPA + ADM + CDDP + 5-FU) to evaluate the sensitivity of these indices to renal tubular damage. After receiving CDDP almost all patients displayed an increase in excretion of beta-m but no urinary enzyme activities. However, NAG index (NAG activity/urinary creatinine) rose markedly in all patients. We concluded that NAG index is a valuable method in providing sensitive indices for detecting renal tubular damage caused by CDDP. PMID- 3044274 TI - [An experimental study on SRCA (subrenal capsule assay)--using immunosuppressive maneuvers]. AB - Six-day SRCA using normal mice developed by Bogden et al. is one of the most promising methods for in vivo chemosensitivity tests. However, this method has a problem on the influence of the host reaction during six days. Therefore, we examined the tumour growth kinetics, host reaction and expression of antitumor effect on three immunosuppressive maneuvers; cyclophosphamide (EX), cyclosporin A (CSA), and total body irradiation (TBI). The tumour diameter increased until day 16 in EX and CSA-treated groups and day 10 in TBI-treated group, but the results of the histological examination showed that tumour cells were preserved in tissue on day 14 in CSA-treated group and day 6 in EX and TBI-treated groups. These results were supported by flow cytometrical analysis. The autoradiogram using 3H TdR showed that labelling index of the tumour cells was not affected by these immunosuppressive maneuvers. From the investigation of the antitumour activity of adriamycin and mitomycin C, it was suggested that the 12-day assay was suitable if nude mice were used in SRCA, and six-day assay, if EX-treated normal mice were used. In CSA-treated group, toxicity of anticancer drugs was manifested than usual. PMID- 3044275 TI - [Phase II study of carboplatin in ovarian cancer]. AB - A phase II study of carboplatin was conducted in ovarian cancer by a cooperative study group consisting of 22 institutions nationwide. The overall response rate of 50 evaluable cases was 38.0%. The response rate in cases with no prior chemotherapy was 54.2% and that with prior chemotherapy was 23.1%. In addition, the response rate was 26.3% in cases that received prior CDDP-based chemotherapy. Histologically, the response rate was high in serous cystadenocarcinoma and endometrioid adenocarcinoma. Hematologic toxicities, particularly leukopenia and thrombocytopenia, were frequently observed, but well tolerated, while renal and neurologic toxicities were rare. These results suggested that carboplatin is useful in the treatment of ovarian cancer. PMID- 3044276 TI - [Phase III study of carboplatin in ovarian cancer]. AB - A phase III study of carboplatin vs cisplatin in CAP regimen for ovarian cancer was conducted by a cooperative study group consisting of 22 institutions nationwide. The response rate for 23 cases allocated to carboplatin regimen group was 34.8% and 42.1% for 19 cases allocated to cisplatin regimen group. No significant difference was observed between these two regimens in response rates, and no significant differences were noted between the two regimens in each of the clinical toxicities and abnormalities in laboratory findings. Since the carboplatin regimen group did not require hydration, or the use diuretics or antiemetics, these results suggested that carboplatin is more useful in the treatment of ovarian cancer than cisplatin. PMID- 3044277 TI - [Phase II study of carboplatin in genitourinary cancer. Urological Cooperative Study Group on Carboplatin]. AB - Carboplatin was evaluated in a phase II study involving 109 patients with genitourinary cancer. A total of 21 cases of advanced testicular tumor, 38 of transitional cell carcinoma (TCC) of the urinary tract and 25 of prostatic cancer were evaluable for response. The response rate in testicular tumors was 48%, with 70% in seminomas and 27% in nonseminomas. Three responses were observed in patients previously treated with cisplatin. In TCC, the response rate was 18%. No response was observed in prostatic cancer. Carboplatin was well tolerated with no significant renal impairment and ototoxicity detected. Nausea and vomiting were experienced by 50% of patients but the severity was low. Severe myelosuppression, thrombocytopenia and leukopenia were observed. In conclusion, carboplatin has demonstrated activity in both testicular tumors and TCC of the urinary tract and is worthy of further study, especially in combination with other active drugs. PMID- 3044278 TI - [Intensive 1-(4-amino-2-methyl-5-pyrimidinyl) methyl-3-(2-chloroethyl)-3 nitrosourea hydrochloride (ACNU) and cryopreserved autologous bone marrow transplantation]. PMID- 3044279 TI - Ferdinando Serri festschrift. PMID- 3044281 TI - Immunopathologic studies in pityriasis lichenoides. AB - Skin biopsy specimens from five patients with pityriasis lichenoides et varioliformis acuta and from six patients with pityriasis lichenoides chronica were studied by direct immunofluorescence and by an immunoperoxidase technique using a panel of monoclonal antibodies. The dermal inflammatory infiltrate was composed of T cells, macrophages, and a small proportion of CD1a+ cells, mostly perivascular. CD8+ cells (cytotoxic/suppressor phenotype) predominated in the epidermis according to the degree of epidermal necroses, whereas CD4+ cells (helper/inducer phenotype) were superior in number among dermal T cells. A few B cells and Leu7+ cells were detected in only a small proportion of lesions. The results obtained confirm that the two conditions are variants of a single disease process and suggest that cell-mediated immune mechanisms may be important in the pathogenesis of the epidermal and vascular damage. Endothelial cells (HLA-DR+ and HLA-DQ+) and CD1a+ cells (epidermal and possibly dermal) could be primarily involved, acting as antigen-presenting cells. PMID- 3044280 TI - A profile of Professor Ferdinando Serri. PMID- 3044282 TI - Blister spreading, a diagnostic test in pemphigus. Loss of epidermal and dermoepidermal cohesion. AB - Differentiation of the pemphigus group of diseases from other bullous dermatoses can be established by spreading of blisters by gentle finger pressure. In combination with histopathology, the same test is diagnostic for every single pemphigus disease. PMID- 3044284 TI - [Relation between chronic gastritis and gastric cancer]. PMID- 3044283 TI - Relationship between dietary intake of organic chemicals and their concentrations in human adipose tissue and breast milk. PMID- 3044285 TI - [Malignant lymphomas and other malignant proliferations in hematopoietic organs in HIV-positive patients]. PMID- 3044286 TI - Chondromas of the hand. A report of thirty-five cases. AB - Twenty-nine patients were treated in the department of orthopedic surgery of Purpan (Toulouse), with thirty-five chondromata of the hand. Follow-up was over one year. Chondroma is a bone tumor predominantly of the left side, second ray of the hand and phalanx. Some multifocal cases have been observed. The diagnosis was made in half the cases through a pathological fracture. Chondroma is characterized by a diaphyseal or a metaphyso-diaphyseal lacuna, wearing away the cortical bone. Treatment consisted of curetting the tumor and filling the cavity with iliac cancellous bone graft. The clinical result was been excellent in 15 cases (60%), good in 6 cases (25%), with only some residual pain, and/or digital swelling. In two cases the finger was stiff, but functionally not disabling. In one case the stiffness was important and very disabling; after several recurrences it evolved into a chondrosarcoma. PMID- 3044287 TI - [2 cases of early "siliconitis" (silicone synovitis)]. AB - Two cases of early silicone synovitis, 12 and 18 months after surgery for partial scaphoid implant arthroplasty (Swanson design) are reported. None of the implants had been fixed by Kirschner wire or sutured to the radius or an other carpal bone. Both implants had been immobilized for six weeks. Initially the result in both cases was good. Pain and loss of range of movement gradually increased, and a large erosive osteolytic defect of the radius was seen on the X-ray after 12 months for the first case and 18 months for the second one. Surgical revision consisted of implant removal, synovectomy with a proximal row carpectomy in the first case and a soft tissue interposition arthroplasty for the second one. The histologic examination showed a foreign body reaction with birefringent material in multinucleated giant cells. All reports about this type of silicone implant complication emphasises the role of compression forces and micro-fragmentation of the silicone. Should we continue to perform partial scaphoid implant arthroplasty from whom microparticles are creating a severe foreign body reaction? PMID- 3044288 TI - [Our experiences with pedicled groin flaps. Apropos of 80 cases]. AB - A series of 80 inguinal pedicle flaps was studied. These flaps were performed at the Aziza Othmana hospital in Tunis for various indications, predominantly for traumatic skin defects of the hand and sequelae of burns. This study evaluated the quality of this auto-plasty, its vascular reliability, its simplicity and the large area of skin cover available. PMID- 3044289 TI - Experimental and clinical study of fast absorption cutaneous suture material. AB - Fast Absorption Vicryl was irradiated with gamma rays. The experimental study on 14 mice grafted with human skin showed a constant rupture of the sutures on the 12th day, with the development of a moderate macrophage reaction. Injection of isotonic saline around the suture material did not accelerate the process of resorption. The clinical study with 27 children operated on the limbs showed rupture of the sutures between the 12th and 16th days, without inflammatory reaction, infection or wound dehiscence. Simply wiping the wound with a compress removes the stitches painlessly. This is appreciated by the young subject and his or her family. Since we began to use FAV systematically we have not needed to conduct dressing under anesthesia. The long term results of healing (follow-up: 11.3 months) are similar to those obtained with conventional non-absorbable suture material. The tolerance, the rapid resorption, the comfort of the patient and the cost advantage make FAV the suture material of choice. PMID- 3044291 TI - [Serum levels of IgG subclasses in the normal child. Evaluation by an immunoenzymatic method using monoclonal antibodies]. AB - Serum IgG subclass levels were measured by an immunoenzymatic assay with monoclonal antibodies in 225 normal children aged 1 to 17 years. Adult concentrations of IgG1 and IgG3 were reached early (2 to 3 years of age), with mean IgG1 level slightly higher in children than in adults, whereas IgG2 and IgG4 showed a slow increase. Mean IgG2 levels in children aged 13 to 17 were still significantly lower than in adults. The distribution of IgG4 levels in every age group was very heterogeneous, with a fair incidence of subthreshold concentrations especially before 5 years of age. These data must be taken into account when defining IgG subclass deficiencies in children. PMID- 3044290 TI - Bacterial NAD(P)-independent quinate dehydrogenase is a quinoprotein. AB - Acinetobacter calcoaceticus LMD 79.41 produced significant amounts of pyrrolo quinoline quinone (PQQ) in its culture medium when grown on quinic acid or shikimic acid. Studies with LMD 79.41 and PQQ- -mutants of this strain demonstrated that this organism contains an NAD(P)-independent quinate dehydrogenase (QDH) (EC 1.1.99.-), catalyzing the first degradation step of these compounds, and that the enzyme contains PQQ as a cofactor, i.e. is a quinoprotein. Synthesis of QDH was induced by protocatechuate and the enzyme appeared to be particle-bound. Acinetobacter Iwoffi RAG-1 produced a quinoprotein QDH apoenzyme since growth on quinic acid only occurred in the presence of PQQ. The results obtained with the PQQ- -mutants of strain LMD 79.41 also provided some insight into the regulation of PQQ biosynthesis and assemblage of quinoprotein enzymes in the periplasmic space. Since two species of Pseudomonas also contained a quinoprotein QDH, it is assumed that bacterial NAD(P) independent quinate dehydrogenase is a quinoprotein. PMID- 3044292 TI - [Neonatal Caffey-Silverman disease with thrombocytosis, increase in C-reactive protein and immunoglobulins]. AB - An unusual case of Caffey-Silverman's disease with thrombocythemia and increased type M immunoglobulins and C-reactive protein levels is reported. These particularities were rarely reported in the literature. These conditions suggest that control should be exercised before steroid treatment in view of the known thrombocythemic effect of the drug. These hematologic abnormalities suggest that this syndrome is infectious in origin and emphasize the risk of steroid treatment. PMID- 3044293 TI - [Iconographic rubric. Shwartzman's syndrome]. PMID- 3044294 TI - [Arterial hypertension in the newborn infant. Etiologies. Treatment]. PMID- 3044296 TI - Construction of acetabular bone stock. A staged cementless arthroplasty of the hip. AB - Vascularized pedicular bone graft was used for the purpose of achieving a sufficient bone stock for cementless total hip arthroplasty in patients with major acetabular bone defects. A useful bed for the acetabular prosthesis was obtained in all three cases. The ordinary prosthetic arthroplasty was performed in the second phase. The vitality of the grafted bone was then controlled. The results after a follow-up time of 23, 32 and 40 months respectively are good. PMID- 3044295 TI - Early recognition of postoperative hematoma formation. AB - Postoperative hematoma formation must be treated as a potential infection. One fourth of all postoperative hematomas are already contaminated. Ultrasonographical examination is an effective method for early recognition of such postoperative hematomas. Ultrasonic diagnosis on a routine basis is not necessary, but it should be carried out as soon as clinical symptoms appear. Our postoperative late results with only one early and one late infection after 100 postoperative treatments of the hip joint and femoral shaft emphasize the importance of early diagnosis of hematomas. PMID- 3044297 TI - Hypogonadotropic hypogonadal men respond less well to androgen substitution treatment than hypergonadotropic hypogonadal men. AB - This research asked whether androgen substitution therapy is as efficacious in hypogonadotropic hypogonadal men as in hypergonadotropic hypogonadal men. Erotosexual functions of two groups of six men of each diagnostic category were compared after 5-6 years of continuous androgen treatment. Treatment regimen was the same in both groups: Parenteral testosterone esters 250 mg/2 weeks. No difference was found in erectile and ejaculatory potency, but the number of sexual acts and scores of subjective quality of sexual acts, sexual excitement, and frequency of sexual thoughts and of nonsexual parameters as vigor, fatigue, anxiety were more negative in the hypogonadotropic men. The most obvious difference between the two groups is the value of LH/FSH and presumably of LHRH. Hypogonadotropic hypogonadal men may be better treated with gonadotropins (or with pulsatile LHRH, when the hypophysis is intact) than with androgens. PMID- 3044298 TI - Sexual dysfunction in the diabetes female: a review. AB - Numerous studies have substantiated the presence of sexual dysfunction related neuropathy in the diabetic male. Studies concerning the sexual dysfunction of the diabetic female are few and inconclusive. Controlled research using objective physiological measures is needed to test the hypothesis that the diabetic female is at equal risk of developing neuropathic-related sexual dysfunction. PMID- 3044299 TI - [The role of intraepidermal macrophages (Langerhans cells) in the structural functional organization of the epidermis]. PMID- 3044300 TI - [Patho- and morphogenesis of diabetes mellitus and early diabetic nephropathy]. AB - Heterogeneity of diabetes mellitus, as the specialists see it nowadays, is determined by the presence of histocompatibility antigens and the onset of immune autoaggression against beta-cells of islet of Langerhans which leads to formation of circulating immune complexes and complexes in situ. This may give rise to diabetic nephropathy. In florid diabetes mellitus there also appeared disturbances induced by metabolic derangement, relevant antigens being then exogenic insulin and low-molecular lipoproteins entailing drastic changes of the basal membrane of glomerular capillaries, plasma saturation of the vascular walls and eventual hyalinosis and capillary obliteration. PMID- 3044301 TI - The role of the endothelial dependent relaxing factor in the regulation of cerebral circulation. AB - It has recently been demonstrated that vessel dilation induced by several physiological agents is dependent on an intact vascular endothelium. In order to explain this endothelium dependence, it has been hypothesized that a still unknown chemical substance, generically named Endothelium Dependent Relaxing Factor (EDRF) is necessary for physiological vasodilation. The role of this EDRF in the cerebrovascular physiology is not yet well understood. In this article the cerebrovascular physiological control is reviewed in relationship with possible EDRF actions. The importance of endothelial lesions in the cerebrovascular responses is discussed. PMID- 3044302 TI - Pharmacokinetics of aminoglycoside antibiotics in blood, inner-ear fluids and tissues and their relationship to ototoxicity. AB - This review critically evaluates the literature on aminoglycoside pharmacokinetics in order to answer the question how fluid and tissue levels of the drugs relate to the development of ototoxic and nephrotoxic side effects. We will summarize the evidence that: (1) aminoglycosides do not accumulate in inner ear fluids; (2) aminoglycoside levels in fluids do not correlate with the ototoxic potential of a drug, and (3) selective toxicity cannot be explained by selective tissue penetration of the drugs. We suggest that studies of drug disposition at the cellular level after chronic aminoglycoside treatment be conducted to establish whether a cell-specific uptake contributes to the selective toxicity of the aminoglycoside antibiotics. A sequence of biochemical events that may lead to the development of toxicity at the molecular level is briefly described. PMID- 3044303 TI - 1928-1988: ADA's Diamond Jubilee in Australia's bicentennial year. Sixty years. PMID- 3044304 TI - The Brisbane Statistical Division Survey of Adult Dental Health 1984. 2. Sociological aspects of the survey. PMID- 3044305 TI - Posterior edentulousness and the prescription of partial dentures. PMID- 3044306 TI - Re-attachment of original tooth fragment to a fractured crown. Case report. PMID- 3044307 TI - The position of the apical foramen of the permanent incisors. PMID- 3044308 TI - The Brisbane Statistical Division Survey of Adult Dental Health 1984. 3. Dental health status and treatment needs. PMID- 3044309 TI - Co-ordinated treatment of secondary cleft deformities. PMID- 3044310 TI - Professionally defined dental treatment needs of middle-aged and older patients attending 21 general practitioners, as related to tooth retention. PMID- 3044311 TI - Zinc phosphate cement and retention. PMID- 3044312 TI - Adhesion of composite resin to etched glass ionomer cement. PMID- 3044313 TI - Principles of clasp retention: a review. PMID- 3044314 TI - The role of ultrasound in the diagnosis of parenchymal disease in transplanted kidneys. PMID- 3044315 TI - Bladder deformity. A sign of major inferior venous obstruction. Two case reports and a review. PMID- 3044316 TI - Alfred George Fryett. Radiographer. PMID- 3044317 TI - Malignant pleural mesothelioma in a child--a case report. PMID- 3044318 TI - The impact of intravenous digital subtraction angiography on the evaluation of carotid artery disease: review of the first 600 cases. PMID- 3044319 TI - Computed tomography in radiotherapy treatment planning. PMID- 3044320 TI - Myelographic differentiation of spinal cord arteriovenous malformations from the normal population. PMID- 3044321 TI - The R.A.C.R. survey of intravenous contrast media reactions. A preliminary report. PMID- 3044322 TI - An evaluation of precordial ultrasonic monitoring to avoid bends at altitude. AB - Several investigators have reported that intravascular bubbles can be detected in decompressed subjects before they develop bends. The altitude exposures were generally of short duration with a limited number of subjects. This important preliminary finding needed to be verified in a larger sampling of long duration altitude exposures. In this experiment, 32 subjects in 82 flights were taken to 27,500 ft simulated altitude for 8 h or until the subject developed mild but steady joint pain (bends). Many subjects took more than one flight. At altitude, the subjects were monitored for circulating bubbles by a team of well-trained, experienced technicians. It was determined that bubbles, clearly audible even to untrained observers, occurred in 77% of the flights in which the subjects developed bends. On the other hand, no bubbles were found in 61% of the flights in which the subjects remained bends free even though the subjects were monitored by more than one experienced technician. Therefore, at 27,500 ft ultrasonic monitoring will miss about 25% of the subjects who developed bends (false negatives) and will incorrectly identify a little less than half of the subjects who do not develop bends as potential benders (false positives). PMID- 3044324 TI - Alcohol, aviation, and safety revisited: a historical review and a suggestion. AB - A 1964 study by Harper and Albers initiated a new era of alcohol, aviation, and safety interest. Studies by the author agreed with the Harper and Albers data. Educational efforts that followed, and the apparent effects, are described. A study commissioned by the author confirmed the effect of low levels of alcohol on flying performance. Impairment from other effects of alcohol, such as alcohol induced hypoglycemia (AIH), post-alcohol impairment (PAI), and positional alcohol nystagmus (PAN) are mentioned. It is suggested that the effects of PAN may play a role in spatial disorientation. It is also suggested that the ingestion of alcohol days before an accident may be demonstrated by laboratory procedures. The feasibility and implementation of this should be established. If the association between previous alcohol ingestion, and spatial disorientation or other impairment becomes evident, it should provide a basis for extensive education regarding aviation and safety. PMID- 3044323 TI - The Republic of Singapore Navy's Scopoderm TTS study: results after 2,200 man days at sea. AB - Most clinical trials investigating prevention of seasickness with transdermal scopolamine have been short-term, and little information regarding long-term use is available. We report here a double-blind trial conducted on board a flat bottomed Republic of Singapore Navy vessel of 2,490 tons, sailing in the South China Sea. The trip lasted 26 d, but self-assessment by the 122 adult male participants (using a simplified scale quantifying the level of seasickness) was confined to the 18 d spent at sea. We found that the protection rate with transdermal scopolamine was 46-57%, with maximum benefits to inexperienced participants, during the early preadaptation phase, or in rough seas even after adaptation. Unwanted effects were few and generally minor. We concluded that the benefits of Scopoderm were worth exploiting even for long trips in moderate-to rough seas. PMID- 3044325 TI - Resolutions of the Aerospace Medical Association from 1929-41: Part II--1934-36. AB - The resolutions of the Aerospace Medical Association (formerly the Aero Medical Association of the United States) have been obtained and reviewed in detail for the period 1929-41. They provide a perspective on what Association members considered priority matters during the period. The resolutions cover subjects such as pilot fatigue, aviation medical examiner selection, military flight pay for flight surgeons, and other important issues, including medical standards. These resolutions had a significant impact during the early growth of civil aviation, and a number still influence today's aeromedical practice as well as certain flight operations. PMID- 3044326 TI - Cell biology of bone. PMID- 3044327 TI - Management of osteoporosis. AB - Osteoporosis is clearly more easily prevented than treated. Prevention should start early for those thought to be most at risk. However, for the majority, menopause is the most convenient time point at which to evaluate the necessity for prevention. While oestrogens are the most effective therapeutic agent for prevention of postmenopausal bone loss, they must be used judiciously with attention to reduction in other nutritional and lifestyle risk factors. When given to women with an intact uterus, oestrogen prescription currently should be accompanied by progestogens, several of which may also reduce bone loss. All individuals should be encouraged to obtain an adequate calcium intake (100-1500 mg day-1) and to continue an active lifestyle. Therapy of the clinically overt disorder follows similar guidelines, with therapeutic options including calcitonin androgens and diphosphonates as well as oestrogens as antiresorptive agents. Methods of stimulating new bone formation are limited, the best documented effects being those of fluoride. However, care must be taken in its use and efficacy has yet to be finally established. Other modalities are as yet experimental. Much can be gained by careful attention to the general health of the patients and their environment, since reduction in risk of falls, and thus fracture, is at least as important as attempts to modify the skeleton. PMID- 3044328 TI - Osteomalacia. PMID- 3044329 TI - Renal osteodystrophy. AB - Over the past decade important advances in our understanding of the pathophysiology and treatment of renal osteodystrophy have been made. In particular, the role of calcitriol deficiency in the genesis of hyperparathyroidism in early renal failure is now better understood. So too are the effects of aluminium on bone, and whereas the more florid aluminium related disease is now unusual the more subtle effects of aluminium are now being appreciated. There is still a major problem in the long-term treatment of hyperparathyroid bone disease. The reasons why parathyroid gland proliferation continues to occur on dialysis therapy require a better understanding of cellular events regulating hormone production and parathyroid cell replication. The case for early intervention with vitamin D is now strong but whether such an approach materially influences the long-term outcome is not yet established. Changes in the approach to treatment and in the modalities used for renal replacement therapy will continue to modify the nature of the bone disease. PMID- 3044330 TI - Osteogenesis imperfecta. AB - Major advances have occurred in the classification of OI and in the definition of underlying molecular defects. A clearer understanding of the pathogenesis of OI and of the relationships between the phenotypes and genotypes should emerge. The study of induced mutations in selected regions of the collagen genes with expression in cultured cells or transgenic mice should hasten this process. These advances will also provide a basis for studies into the large number of other genetically determined connective tissue disorders that are grouped together as the skeletal dysplasias. The results of recent studies in OI are providing a unique insight into many aspects of collagen and connective tissue biochemistry, physiology and pathology. PMID- 3044331 TI - Paget's disease of bone. AB - Paget's disease is a common disorder of bone of presumed viral aetiology which, in a minority of patients, may cause severe symptoms. Both calcitonin and the diphosphonates have specific anti-pagetic effects. Newer diphosphonates, unlike etidronate, do not impair mineralization, and when they become available these will be the drugs of choice. Until that time, both calcitonin and etidronate continue to have a useful place in the management of this disorder. PMID- 3044332 TI - Uses and limitations of bone biopsy in management of metabolic bone disease. AB - Bone biopsy is now an established diagnostic and investigative procedure which is widely used in the management of metabolic bone diseases. Its unique feature is that it facilitates direct visualization of bone cells and bone structure and, through double tetracycline labelling, allows the indirect measurement of bone turnover. Its main diagnostic contribution has been in unravelling the complexities of renal osteodystrophy, which consequently is now much better understood. However, it has also uncovered a variety of mineralization defects caused by agents such as fluoride, diphosphonates and aluminum that might otherwise have bone unsuspected clinically. Its main investigative roles have been to provide unique insight into the mechanisms of bone loss with age and in osteoporosis, and to assess the effects on bone and bone cells of potential therapeutic agents in established osteoporosis. This has been possible through the use of histomorphometry, which is used to quantify changes in bone volume and turnover as well as alterations in the microanatomical structure of bone that can reduce bone strength and increase the risk of fracture. Although most biopsies will continue to be taken for histomorphometry, recent studies have shown that specimens may be used to provide additional information such as quantitation and localization of elements within bone, and to provide bone cells for in vitro culture. It is likely that, in future, specimens will be taken from patients with various disorders so that the techniques of cell and molecular biology can be applied to harvested bone cells and thus provide a new dimension to our understanding of metabolic bone disease. PMID- 3044333 TI - Bone scanning and photon absorptiometry in metabolic bone disease. AB - Bone scan imaging with the current bone-seeking radiopharmaceuticals, the technetium-99m labelled diphosphonates, provides a functional display of skeletal metabolism and has dramatically improved our ability to investigate skeletal pathology. While the role of bone scanning in metastatic disease was recognized early, and welcomed by oncologists and other specialists alike, in the field of metabolic bone disease the realization of its potential value has been slower, with perhaps more reluctant acceptance. Nevertheless, the skeleton remains an extremely difficult organ to investigate, and a familiarity with bone scanning should be an essential part of the investigational armamentarium for anyone with an interest in the metabolic bone disorders. The measurement of bone mineral by photon absorptiometry is a relatively new technique, which is rapidly gaining acceptance. The ability to accurately detect and quantify changes in bone mass is potentially of considerable value in the diagnosis and management of osteoporosis. The technique provides us with an extremely important and powerful research tool but, at the present time, its role in clinical practice has yet to be established. PMID- 3044334 TI - Osteoporosis: pathogenesis and risk factors. PMID- 3044335 TI - Diagnostic and therapeutic trends in blood coagulation and fibrinolysis. PMID- 3044336 TI - [Crystallized human plasma proteins. I. Albumin, transthyretin, retinol-binding protein, ceruloplasmin and beta 2-glycoprotein I]. AB - The presently known possibilities and conditions for achieving the crystallization of Albumin, Transthyretin, Retinol-binding Protein, Ceruloplasmin and beta 2-Glycoprotein I are described with respect to our own experiences. For isolating some proteins the crystallization gives rise to a real purification step. Sometimes degrees of purity near to 98 till 99% will be reached. Furthermore crystalline proteins are very important for the determination of the three-dimensional structure by aid of the X-ray diffraction analysis. For this purpose large single crystals are needed with minimum requirements of 0.4 mm length for all the three dimensions. Additionally the crystals must have a high degree of order on the molecular level and should allow recording of the diffraction pattern out to 3 A resolution. From the five proteins described here the three dimensional molecular structure of Transthyretin and Retinol-binding Protein could be elucidated by using the method of the X-ray diffraction analysis. PMID- 3044337 TI - Monoclonal antibodies directed to leukocyte differentiation antigens for therapeutic use. AB - During the past years monoclonal antibodies directed to human leukocyte differentiation antigens have resulted not only in fundamental new knowledge of function and differentiation of leukocyte-populations, but have also turned out to be excellent and new tools for the treatment of lymphoid malignancies. Most widely and successfully monoclonal antibodies directed to T-cell differentiation antigens have been used for immunosuppression in organ transplantation. In the present paper, experimental and clinical data are analyzed which may give information about the criteria for clinical effectivity of distinct monoclonal antibodies. The conclusions clearly show, that not only specificity of an antibody, e.g. recognition of a distinct antigen, but also the recognized epitope, affinity and isotype of an antibody play a crucial role for the biological effectivity of a monoclonal antibody. Clinical success in antibody therapy will therefore be dependent on the possibilities to alter antibody molecules according to the special clinical situation. PMID- 3044338 TI - Anti-idiotypic antibodies: powerful tools in diagnosis and therapy. AB - Mouse x mouse hybridomas secreting monoclonal anti-idiotypic antibodies have been prepared from BALB/c mice immunized with syngeneic monoclonal antibodies specific for IgE, AFP, CEA and TSH respectively. The hybridomas were selected on the basis that the secreted antibodies competed with antigen for binding to the immunizing idiotope. Using IgE and AFP as a model it could be shown that syngeneic antigen inhibitable monoclonal anti-idiotypic antibodies can act well as antigen surrogate. Thus a complementary idiotype-anti-idiotype antibody pair could be used in a competitive assay wherein the extend of idiotype-anti-idiotype complex formation is inversely proportional to the amount of antigen in the sample. Apart from this a lot of human and animal studies have shown that this interaction between idiotype and anti-idiotype could be utilized to prevent certain infectious diseases and to treat some kinds of cancers. PMID- 3044339 TI - Investigations with monoclonal antibody drug (anthracycline) conjugates. AB - The development of monoclonal antibody-drug (anthracycline) conjugates (immunocytostatics) that has emerged during the last decade is briefly reviewed. Stress is layed on the various procedures that have been employed for the chemical attachment of daunorubicin (daunomycin) and doxorubicin (adriamycin) to spacers, high-molecular weight carrier molecules, and immunoglobulins. Anthracycline conjugates that have proved effective in mice with respect to the treatment of cancer are especially evaluated. Also a new method developed for the conjugation of rhodosaminyl anthracyclinones via a hydrolysable spacer, developed in our laboratories, is briefly communicated. Finally, import aspects for further developments of monoclonal antibody-drug conjugates are discussed. PMID- 3044340 TI - 15-deoxyspergualin: from cytostasis to immunosuppression. AB - This article gives a survey on the pharmacological action of the new polyamine compound 15-Deoxyspergualin (15-DS). 15-DS is a derivative of the antitumor antibiotic spergualin which has been isolated by Prof. Umezawa's group in Japan. Besides its antitumor activity 15-DS was shown to possess immunosuppressive properties making the substance suitable for the application in organ transplantation and autoimmune disorders. Due to its immunopharmacological mode of action known hitherto 15-DS clearly differs from other immunosuppressants as e.g. Cyclosporin A. PMID- 3044341 TI - Efficient expression system for human antithrombin III in baby hamster kidney cells. AB - A DNA fragment carrying the enhancer of an immediate early gene of human cytomegalovirus (hCMV) was tested as a transcriptional control element by fusion downstream of a transcription unit for human antithrombin III (AT III) driven by the Simian virus (SV40) enhancer and promoter. Measurement of transient AT III expression in baby hamster kidney cells (BHK) shows that by the presence of the hCMV enhancer the synthesis of AT III is increased considerably (three to fourfold). The AT III expression vectors carrying the hCMV enhancer were used to establish stable BHK cell-lines by a new and fast G418/methotrexate selection protocol, which express up to 12 micrograms AT III/10(6) cells/24h after 40-50 days. The given system might be generally useful for the fast expression of recombinant proteins in mammalian cells, e.g. the screening of altered AT III molecules obtained by site directed mutagenesis. PMID- 3044343 TI - A histidin alanine rich recombinant antigen protects Aotus monkeys from P. falciparum infection. AB - We have isolated a cDNA clone coding for 165 amino acids of a histidine alanine rich protein. An extended repeat region of this clone codes for the tripeptide Ala-His-His, which is extremely conserved, and for the tripeptide Ala-Ala-Asp, which shows only slight variability among the repeat units. This antigen exhibits high homology to the HRPII antigen described by Wellems and Howard (1986). The coding region was expressed in E. coli as a MS2-polymerase fusion protein, which was purified and used for immunization of Aotus monkeys. The animals immunized with this fusion protein showed only low parasitemias (less than 2%) after infection with P. falciparum, while animals from the control group or animals immunized with a MS2-fusion protein carrying other malaria specific sequences were not protected. The result suggests that this antigen is a good candidate for a malaria vaccine. PMID- 3044342 TI - Immunoreactivity of human immunodeficiency virus (HIV-1) envelope polypeptides expressed in Escherichia coli. AB - Defined cDNA sequences encoding segments of envelope glycoproteins of the human immunodeficiency virus were expressed in E. coli. The recombinant env fusion proteins were tested for immunoreactivity with patient sera by radio immunoprecipitation procedures. The regions of the env proteins, carrying relevant antigenic determinants in respect to diagnostic purposes, could be identified. PMID- 3044344 TI - Modern trends in the development of bacterial vaccines for human use. AB - The development of vaccines against infectious diseases is one of the great medical achievements of the 20th century. What other medical measure had such an influence on the statistical prolongation of life, as the use of vaccines did? The successful development of today's vaccines was from the beginning due to the intensive cooperation between the basic research sciences (e.g. bacteriology virology, chemistry, immunology) and the applied sciences (e.g. medicine practiced). During the last few years newly developed techniques have awakened hopes for the development of new vaccines which are better tolerable and more effective. Molecular biologic methods have shown first successes in the development of epitope vaccines. Monoclonal anti-idiotype antibodies are being used in active protection experiments. Modern galenic preparations can perhaps one day replace the parenteral administration of antigens. In the following some of the new techniques are listed and their significance for the development of new vaccines discussed. PMID- 3044345 TI - Factor XIII: enzymatic and clinical aspects. AB - The present knowledge about clotting F XIII is roughly displayed. It is explained why F XIII is not a clotting factor in the strict sense of the word and that its biological function more general is the crosslinking of proteins via a transamidase reaction, the reduction of the permeability of the microvasculature, the stimulation of connective tissue cells, and finally the steering of connective tissue processes involved in wound healing. These effects are demonstrated as well in cell culture studies, animal experiments as by clinical results. Concerning the clinical results with F XIII, its effects on sclerodermia, subarachnoidal hemorrhage, inflammatory bowel diseases, Purpura Schonlein Henoch, and the wound healing are described in more detail. PMID- 3044346 TI - Recent advances in the biochemistry of the fibrinolytic system. AB - In the last decades, tremendous advances have been made in the elucidation of the biochemical mechanism of the fibrinolytic system. The activators of plasminogen have been accurately characterized and the mechanism of the activation reaction studied in great details. Recently four new inhibitors of the plasminogen activators have been identified, their biochemical and physiological properties are under study. Although the various roles of plasmin, only its function in the lysis of fibrin will be considered in this review. PMID- 3044347 TI - Inactivation of AIDS-causing retroviruses by the manufacturing procedures for human plasma proteins. AB - Human retroviruses causing AIDS (HIV) may occur in human plasma. Since HIV contaminated plasma cannot be completely excluded by testing for anti-HIV, AIDS safety of human plasma products can only be achieved by introducing HIV inactivating and/or eliminating methods into the manufacturing procedure. Here we review a number of methods used when manufacturing plasma derivatives at Behringwerke. These methods were previously developed either to produce a protein of required purity or to manufacture hepatitis safe products. Methods used to produce purer proteins are ethanol fractionation, pepsin treatment, affinity chromatography or various protein precipitation procedures. The method developed at Behringwerke for inactivating infectious viruses in plasma protein preparations not destroying the biological activities of the human protein is pasteurization, i.e. 10 h heat treatment of the aqueous protein solution at 60 degrees C. To investigate the HIV inactivating efficiency of the methods mentioned above, aliquots of an infectious HIV type 1 concentrate were added to a protein preparation, the resulting HIV spiked preparation treated according to the method to be studied and the amount of infectious HIV in this preparation determined before and after treatment. By all methods reported on here the HIV type 1 isolate was completely inactivated resulting in high inactivation factors. In addition, the heat stability of HIV type 2 was tested in aqueous solution at 60 degrees C proving that both HIV-1 and HIV-2 isolates are of comparably low heat stability under these conditions. From the results discussed here it can be concluded that all commercial human protein products of Behringwerke derived from either human plasma or placenta do not contain any infectious retrovirus causing AIDS and thus have a high margin of safety regarding the transmission of AIDS. PMID- 3044348 TI - Preparation and use of synthetic blood group specific immunoadsorbents. AB - The carbohydrate structures determining the ABO blood group system have been almost completely characterized. Recent advances in the stereoselective synthesis of such complex structures have made it possible to prepare pure oligosaccharides in large quantities and to study their possible uses for diagnostics and pharmaceutical processing techniques. In the experiments described here A and B blood group specific determinants were synthesized and bound to solid phases by means of suitable spacers in order to study their use as immunoadsorbents. The objective was to adsorb blood group specific antibodies from positive sera and from immunoglobulin preparations. It was shown that the anti-A and anti-B antibodies bound to the immunoadsorbents with high affinity could be removed effectively. The effects were achieved both using affinity chromatography and "batch processing". PMID- 3044349 TI - The relationship between panic, phobic and anticipatory anxiety in agoraphobia. PMID- 3044350 TI - A refined switch-activated time monitor for the measurement of sleep-onset latency. PMID- 3044351 TI - [Neonatal hip screening using sonography]. PMID- 3044352 TI - [Achilles tendon injury--a historical retrospect]. PMID- 3044353 TI - Concordant divergence in proteins and mitochondrial DNA between two vole species in the genus Clethrionomys. AB - The bank vole (Clethrionomys glareolus) and the northern red-backed vole (C. rutilus) are two closely related species where interspecific crosses result in fertile female but sterile male offspring. Mitochondrial DNA (mtDNA) from C. rutilus has passed the species barrier and is found in C. glareolus from northern Fennoscandia. The present report shows that the genetic distance between the two species, calculated from enzyme data (Nei's D), is 0.64. Isoelectric focusing of muscle proteins resolved around 55 bands, of which each species had 6 or 7 bands not present in the other species. Sequence divergence of mtDNA from the two species is 13.9%. A comparison between protein and mtDNA distances in other species pairs reveals a high correlation between the two measures, indicating that differences in mtDNA between taxa are not random when compared to divergence in protein-coding nuclear genes. The relationship between genetic divergence in proteins and that in mtDNA between Clethrionomys glareolus and C. rutilus is similar to that found in other species pairs. It is also shown that despite large differences on the protein level it is still, in some cases, possible for species pairs to produce fertile hybrid females. PMID- 3044355 TI - Megabase methods: a quantum jump in recombinant DNA techniques. PMID- 3044354 TI - Genetic analysis of Triticeae shikimate dehydrogenase. AB - Starch gel electrophoresis and polyacrylamide gel isoelectric focusing (IEF) were used to investigate the genetic control of Triticeae shikimate dehydrogenase-1 (SKDH-1). Studies of wheat-alien species chromosome addition lines established that Skdh-1 of Hordeum vulgare cv. Betzes is located in chromosome 5H, Skdh-V1 of Dasypyrum villosum in 5V, Skdh-R1 of Secale cereale cvs. Dakold and King II in 5R, and Skdh-S1(1) of Triticum longissimum in 5S1S. Also, the chromosomal locations of the genes that encode SKDH-1 in T. aestivum cv. Chinese Spring, T. umbellulatum, and S. cereale cv. Imperial, determined earlier using zone electrophoresis, were reconfirmed using IEF. Zone electrophoresis and IEF do not differ markedly in their ability to detect the expression of alien Skdh-1 genes in wheat-alien species chromosome addition lines. However, IEF may be superior to zone electrophoresis as a technique for detecting and analyzing SKDH-1 genetic variants within Triticeae species; among the species studied, IEF generally resolved two or more isozymes per Skdh-1 allele present, while zone electrophoresis resolved only one. PMID- 3044356 TI - Early embryogenesis in Caenorhabditis elegans: the cytoskeleton and spatial organization of the zygote. PMID- 3044357 TI - Neuropeptides, second messengers and insect molting. PMID- 3044359 TI - Facts and hypotheses concerning the function of non-granulated cells in the adenohypophysis of vertebrates. PMID- 3044358 TI - Translational regulation in sea urchin eggs: a complex interaction of biochemical and physiological regulatory mechanisms. PMID- 3044360 TI - Pre-implantation diagnosis. PMID- 3044361 TI - Bacterial RNA polymerase--the ultimate metabolic sensor? PMID- 3044362 TI - Molecular and cellular biology of malaria. PMID- 3044363 TI - A conserved eukaryotic cell cycle control. PMID- 3044364 TI - On the architecture of regulatory systems: evolutionary insights and implications. PMID- 3044365 TI - Insulin-mediated post-transcriptional regulation of hepatic malic enzyme and albumin mRNAs. AB - Livers of insulin-treated diabetic rats accumulate albumin and malic enzyme mRNAs at very different rates. We now report that in normal rats insulin directs a specific increase in malic enzyme mRNA, while albumin mRNA levels remain unaltered. These studies support the contention that insulin regulates the accumulation of hepatic mRNAs in a highly specific manner. To evaluate whether or not albumin and malic enzyme mRNA levels are determined by altered rates of transcription, in vitro transcription assays were performed. The results of these studies demonstrate that increased malic enzyme mRNA levels in insulin-treated normal rats and increased malic enzyme and albumin mRNA levels in insulin-treated diabetic rats do not involve altered rates of transcription of the genetic sequences encoding these proteins. For these two specific proteins, insulin mediates changes in mRNA levels by a post-transcriptional mechanism. PMID- 3044367 TI - Ras oncogene mediated induction of a 92 kDa metalloproteinase; strong correlation with the malignant phenotype. AB - We have previously demonstrated that activated ras oncogenes can induce the metastatic phenotype and type IV collagenolytic activity in NIH/3T3 cells (Thorgeirsson et al. Mol. Cell. Biol. 5:259-262, 1985). The present study demonstrates ras-mediated induction of a 92 kDa metalloproteinase, which degrades gelatin and type IV collagen. Association of the 92 kDa proteinase expression with the malignant phenotype was also observed in human tumor cell lines. Our data indicate that the 92 kDa gelatin-collagen IV degrading metalloproteinase is an important participant in the proteolytic process involving tumor cell invasion and metastasis. PMID- 3044366 TI - Effects of insulin and phorbol esters on diacylglycerol generation and synthesis and hydrolysis of phosphatidylcholine in BC3H-1 myocytes. AB - Insulin was found to provoke simultaneous, rapid, biphasic increases in [3H]choline-labeling of phosphatidylcholine and phosphocholine in BC3H-1 myocytes. Phorbol esters increased [3H]choline-labeling of phosphocholine, but not phosphatidylcholine. Both agonists increased diacylglycerol production. These results suggest that: (a) insulin provokes coordinated increases in the synthesis and hydrolysis of PC; and, (b) insulin-induced activation of protein kinase C may activate a PC-specific phospholipase. PMID- 3044368 TI - Cellular mechanism of action by a novel vasoconstrictor endothelin in cultured rat vascular smooth muscle cells. AB - Specific binding sites for synthetic porcine endothelin (pET), a novel potent vasoconstrictor peptide isolated from the supernatant of cultured porcine endothelial cells, and its effects on cytosolic free Ca2+ concentrations ([Ca2+]i) and phosphatidylinositol (PI) response were studied in cultured rat aortic vascular smooth muscle cells (VSMC). Binding of 125I-labeled-pET to rat VSMC was time- and temperature-dependent and the cell-bound 125I-labeled-pET was resistant to dissociate. Scatchard analysis of binding studies indicated the presence of a single class of high-affinity binding sites: the apparent Kd was 2 4 X 10(-10) M and the maximal binding capacity was 11,000-13,000 sites/cell. The binding was highly specific for pET because neither well-recognized vasoconstrictors, peptide neurotoxins, nor Ca2+-channel blockers affected the binding. pET dose-dependently (10(-9)-10(-7) M) induced a transient and sustained increase in [Ca2+]i in fura-2-loaded cells of which effect was largely dependent on extracellular Ca2+, whereas it had no significant effect on PI response in 3H myoinositol-prelabeled cells. The present data clearly demonstrates the presence of specific receptors for pET distinct from those of the well-recognized vasoconstrictors and voltage-dependent Ca2+-channels in cultured rat VSMC, and suggest that pET-induced increase in [Ca2+]i is involved in the mechanism of its vasoconstriction. PMID- 3044369 TI - Acetylated HMG1 protein interacts specifically with homologous DNA polymerase alpha in vitro. AB - The acetylated, deacetylated and nonacetylated forms of HMG1 proteins from Guerin ascites tumour cells and calf thymus were separated and their in vitro interactions with homologous and heterologous DNA polymerases were studied. It has been found that only the acetylated form of HMG1 proteins forms a specific complex with homologous DNA polymerase alpha and stimulates its activity in vitro. The acetylation therefore is necessary for their possible function in DNA replication. This finding represents an evidence for a relationship between the acetylation of HMG1 proteins and their biological role. PMID- 3044370 TI - Effects of the detergent sucrose monolaurate on binding of amphotericin B to sterols and its toxicity for cells. AB - Amphotericin B (AmB) is a potent antifungal agent used to treat patients with systemic mycoses. The cytotoxicity of AmB is related to its binding to membrane sterols and its clinical usefulness is based on its greater affinity to ergosterol, the fungal sterol, compared to the mammalian cell sterol, cholesterol (1-3). Here we report that sucrose monolaurate (L.S.) decreased the binding of AmB to cholesterol without interfering with its binding to ergosterol. Furthermore, the toxicity of AmB for mouse erythrocytes (RBC) and cultured mouse fibroblasts, L-929, cells was significantly decreased by low concentrations of L.S., whereas under the same conditions, its toxicity for Candida albicans was unaffected. We observed a very good correlation between the spectroscopic and cell studies. The results reported here on the effects of L.S. on the selectivity of AmB toxicity for fungal cells compared to animal cells and the relative nontoxic nature of sugar esters suggest a potential for compounds of this type to enhance the therapeutic index of AmB. PMID- 3044371 TI - Relationships between reversion of hydroxyurea resistance in hamster cells and the co-amplification of ribonucleotide reductase M2 component, ornithine decarboxylase and P5-8 genes. AB - Hydroxyurea is a specific inhibitor of ribonucleotide reductase, which is a rate limiting enzyme activity in DNA synthesis. Cells selected for resistance to hydroxyurea contain alterations in ribonucleotide reductase activity. An unstable hydroxyurea resistant population of hamster cells has been used to isolate a stable drug resistant cell line, and two stable revertant lines with different sensitivities to hydroxyurea cytotoxicity and different ribonucleotide reductase activity levels. We show for the first time that a decrease in hydroxyurea resistance is accompanied by a parallel decline in gene copies for the M2 component of ribonucleotide reductase, ornithine decarboxylase and a gene of unknown function called p5-8, indicating that the co-amplification of the three genes is associated with drug resistance, and supporting the concept that M2, ornithine decarboxylase and p5-8 are closely linked, and form part of a single amplicon in hamster cells. PMID- 3044372 TI - Short-term stimulation by insulin of lipoprotein lipase secretion in adipose cells. AB - The spontaneous secretion of lipoprotein lipase has been examined in adipose cells of mouse Ob17, Ob17SA and 3T3-F442A clonal lines as well as in rat adipose cells in primary culture. Striking differences are observed both in serum-free and serum-supplemented media, rat adipose cells and 3T3-F442A cells being the most active. Insulin from 10(-11) M to 10(-9) M was able to modulate the rate of LPL secretion from 2- to 4-fold. The stimulatory effect of insulin on this process occurred within 30 min in cells treated or not with cycloheximide. It is concluded that insulin is able to modulate the rate of LPL secretion independently of the synthesis of new enzyme molecules on a short-term basis and within a physiological range of concentrations. PMID- 3044373 TI - The treatment of Wegener's granulomatosis with trimethoprim/sulfamethoxazole: illusion or vision? PMID- 3044374 TI - The efficacy and safety of auranofin in the treatment of juvenile rheumatoid arthritis. A long-term open study. AB - The safety and efficacy of auranofin in the long-term treatment of children with juvenile rheumatoid arthritis was investigated in an open study of 14 patients. Twelve patients completed at least 12 months of treatment, and 7 patients completed 36 months of treatment. Classic parameters of disease activity showed improvement over baseline values after 6 months of treatment, and laboratory indices remained stable or improved throughout the study. Auranofin was well tolerated; the frequency of adverse effects was lower in these patients than has been previously reported in either adults or children whose arthritis has been treated with injectable gold. PMID- 3044375 TI - Ultrasound in abdominal trauma: an alternative to peritoneal lavage. AB - Ultrasonography has been used as an investigation in patients with abdominal signs after blunt injury. Thirty-two patients were examined, of whom 11 had abnormal findings. Free intraperitoneal fluid was demonstrated in eight cases, seven of whom had this confirmed at subsequent laparotomy. The 21 patients with normal scans did not require abdominal intervention. It is suggested that ultrasonography is a reliable method of detecting haemoperitoneum and offers a valuable non-invasive method of investigating blunt abdominal injuries. The hepatorenal pouch is the site where free intraperitoneal fluid can be most easily demonstrated. PMID- 3044376 TI - Toward earlier diagnosis of splenic injury? AB - A series of patients who were found at operation to have sustained splenic rupture is described and their immediate presenting features are detailed. Signs of peritoneal irritation were not always present and patients were not often 'shocked' when first seen. Helpful early signs included a low haemoglobin and pallor. There is a tendency to underestimate the significance of left quadrant pain in the presence of rib fractures. Peritoneal lavage and ultrasound should be more readily employed. Text book features should not be expected early and this must be taught to junior doctors who work in accident and emergency medicine. PMID- 3044377 TI - Tetanus booster every 5 years: an unnecessary routine? AB - The various guidelines for the administration of tetanus toxoid and antitetanus immunoglobulin are not only complicated, but also have never been supported by any scientific experimental studies. This study has measured the antibody levels in a random sample of 157 patients presenting to an accident and emergency department. Levels were measured before and after boosting doses. The results show that, in the sample analyzed, even those patients who had their last 'booster' over 10 years ago, had a satisfactory immunoglobulin level. In fact, no patient in the study had a level below the 'protective level'. Bearing in mind the small number of patients in the study, it could be argued that the level of immunity against tetanus in the United Kingdom is likely to be higher than assumed. If this is proven to be correct, then the length of time between booster injections of toxoid can be extended and the use of Human Antitetanus Immunoglobulin can be further restricted. PMID- 3044378 TI - Lower limb skin loss: simple outpatient management with meshed skin grafts with immediate mobilization. PMID- 3044379 TI - Computer-unaided diagnosis of acute abdominal pain in an accident and emergency department. AB - The initial accident and emergency and final in-patient diagnoses of 200 patients with acute abdominal pain made without computer aid or structured data sheet were compared. Sixty-five per cent were correctly diagnosed and 5% had normal laparotomies. These results compare favourably with those obtained using computer aid and structured history-taking forms. It is suggested that the spotlight should be on the training and experience of doctors making the initial diagnosis rather than on computer aid. PMID- 3044380 TI - Ultrasound assessment of the suspected scaphoid fracture. AB - A prospective study of 111 patients thought to have sustained a recent scaphoid fracture on clinical grounds but who were radiologically negative was undertaken over a period of 7 months. All such patients were subjected to ultrasound scanning within a week of their injury under double blind conditions. All patients were re-X-rayed 2-3 weeks after their injury. The authors' results suggest that ultrasonic diagnosis of the possibly fractured scaphoid is unreliable. PMID- 3044381 TI - Hormone receptor analysis in endometrial cytologic samples collected with the Gynoscann device. AB - Eighteen patients underwent endometrial cytologic sampling using the Gynoscann instrument immediately before curettage. The material obtained was used for cytologic diagnosis and for measurements of estrogen receptors (ERs) and progesterone receptors (PRs). The material collected with the Gynoscann device provided conclusive cytologic diagnoses in 88% of the cases, as compared with the histologic classification of samples collected by curettage. The DNA content in the samples obtained by the Gynoscann device varied from 5 micrograms to 218 micrograms; this amount was sufficient for the analysis of both ER and PR. Thus, assays for steroid receptors may be performed on endometrial cytologic samples obtained by the Gynoscann; this method may be especially useful for cases in which it is difficult to take a surgical biopsy. PMID- 3044382 TI - Development and use of a rule-based pathology expert consultation system. AB - Since histologic sections of pathologically changed tissues (e.g., tumors and inflammations) are highly complex structures, their subjective diagnostic or prognostic interpretation may result in a lack of interobserver and intraobserver agreement. Rule-based expert systems have the advantages that they require precise representation of the problem and can be used quickly and efficiently, even if the user is not acquainted with the special field concerned. Because of these and other advantages, a rule-based "Pathology Expert Consultation System" (PECS) is under development for the assessment of different premalignancies, malignancies and noncancerous conditions. In addition to supporting diagnosis making, the expert system can infer stage, type and grade, can check for inconsistencies in the answers and can predict the prognosis of an individual patient. PECS permits total integration of information contained in database, worksheet and text files with the rules and other control structures inherent in the rule base. The consultation can be interrupted to display a graph; when the user dismisses the graph, the consultation will resume where it left off or will move to another part of the system (if desired). If the user asks why a question is raised, the system will answer with a textual, graphic or combined explanation. The construction and use of the PECS expert system is discussed. Its initial applications to tumors of the lung, breast and endometrium have shown it to have both educational and clinical significance. PMID- 3044384 TI - [The importance of subject review articles]. PMID- 3044383 TI - Diagnostic schemes for fine needle aspirates of breast masses. AB - A comparison was made of four statistically based schemes for classifying epithelial cells from 243 fine needle aspirates of breast masses as benign or malignant. Two schemes were computer-generated decision trees and two were user generated. Eleven cytologic characteristics described in the literature as being useful in distinguishing benign from malignant breast aspirates were assessed on a scale of 1 to 10, with 1 being closest to that described as benign and 10 to that described as malignant. The original computer-generated dichotomous decision tree gave 6 false negatives and 12 false positives on the data set; another tree generated from the current data improved performance slightly, with 5 false negatives and 10 false positives. Maximum diagnostic overlap occurred at the cut point of the original dichotomous tree. The insertion of a third node evaluating additional parameters resulted in one false negative and seven false positives. This performance was matched by summing the scores of the eight characteristics that individually were most effective in separating benign from malignant. We conclude that, while statistically designed, computer-generated dichotomous decision trees identify a starting sequence for applying cytologic characteristics to distinguish between benign and malignant breast aspirates, modifications based on human expert knowledge may result in schemes that improve diagnostic performance. PMID- 3044385 TI - [Anastomosis of the small-caliber intestine]. PMID- 3044386 TI - [Obstructive sleep apnea]. PMID- 3044387 TI - [Congenital hypertrophy of the pylorus]. PMID- 3044388 TI - Cryopreservation of animal cells. PMID- 3044389 TI - The development of large-scale animal cell perfusion culture systems. PMID- 3044390 TI - Hollow-fiber bioreactors for mammalian cell culture. PMID- 3044391 TI - Microencapsulation of living mammalian cells. PMID- 3044392 TI - Some environmental and physiological factors affecting the sizes of microbial cells. PMID- 3044393 TI - Bioreactor design for growth of shear-sensitive mammalian and insect cells. PMID- 3044394 TI - Myofibroblasts in head and neck surgery. An experimental and clinical study. AB - Myofibroblasts in human granulation tissue have many of the structural and functional characteristics of smooth-muscle cells and appear to be responsible for wound contraction. They have also been identified in contracted scar tissue in nongranulation wounds. In this report, their role in head and neck wound healing will be explored utilizing transmission electron microscopy and immunoperoxidase techniques with antibodies to the intermediate filament vimentin and to muscle-restricted actins. In piglets, high-tension, low-tension, and granulating wounds were created and studied with serial biopsy specimens. Results showed few myofibroblasts in either the high- or low-tension wounds and multiple myofibroblasts in the granulating wounds. In the clinical studies, the immunoperoxidase technique with monoclonal antibody to muscle-specific actins proved most useful in identifying myofibroblasts. Myofibroblasts were present in granulating wounds and hypertrophic scars. They were not widely present in mature keloids. PMID- 3044395 TI - Organelle origins and ribosomal RNA. AB - As the detailed molecular biology of organelle genomes has unfolded, there has been a general acceptance of the view that plastids and mitochondria are of endosymbiotic, eubacterial origin. Plastid genes are strikingly similar to their eubacterial (particularly cyanobacterial) counterparts in sequence, organization, and mode of expression, and such features strongly support the hypothesis that the plastid and its genome were derived in evolution from a blue-green alga-like endosymbiont. Mitochondria, on the other hand, are problematic: mitochondrial genes are organized and expressed in remarkably diverse ways in the different major groups of eukaryotes, and in no case are these features particularly characteristic of either bacterial or nuclear genomes. There is, however, clear evidence derived from gene sequence supporting the eubacterial ancestry of mitochondria, and some of the most compelling data have come from analyses of mitochondrial ribosomal RNA (rRNA). Plant mitochondrial rRNA genes diverge in sequence at a particularly slow rate, and these genes have proven to be especially supportive of the endosymbiont hypothesis, pointing to an origin of mitochondria from within the alpha subdivision of the purple bacteria. Ribosomal RNA sequences provide a basis for the construction of global phylogenetic trees that probe the evolutionary history of organelles, and that address the question of whether mitochondria and plastids are monophyletic or polyphyletic in origin. Such studies raise the possibility that the rRNA genes of plant mitochondria originated separately from the mitochondrial rRNA genes of other eukaryotes. PMID- 3044396 TI - Structure and function of nonhistone phosphoproteins. AB - Many nonhistone nuclear proteins have been shown to be phosphorylated and associated with a diverse range of cellular activities. The aim of this present review is to update the most recent developments in this field with respect to the traditional roles that these proteins have been postulated to play as enzymes, DNA-binding proteins, hormonal receptors, nucleosome associated proteins, and nucleolar associated proteins. In addition, evidence is presented suggesting that these proteins may also function as nuclear oncogene protein products, structural constituents of nuclear organization (e.g., lamina-nuclear matrix associated proteins), and RNA processing factors. PMID- 3044397 TI - Bacterial and mammalian thioredoxin systems activate iodothyronine 5' deiodination. AB - The identity of a dithiol (designated DFB) of relative mass (Mr) = 13,000, reported previously to be present in fraction B of rat liver cytosol and to participate as a cofactor in the 5'-deiodination of iodothyronines, has been investigated. Substitution of highly purified thioredoxin from Escherichia coli for fraction B or of highly purified thioredoxin reductase from either E. coli or rat liver for cytosolic fraction A (containing DFB reductase) permits deiodination of 3,3',5'-[125I]triiodothyronine by rat liver microsomes to proceed. Addition of antibodies to highly purified rat-liver thioredoxin or thioredoxin reductase inhibits deiodination. Thus, the thioredoxin system largely accounts for the activity of the cytosolic cofactor system supporting 5' deiodination of 3,3',5'-triiodothyronine in rat liver. PMID- 3044398 TI - Contaminating coagulase-negative staphylococci isolated in a lysis-centrifugation (Isolator) blood culture system. Application of different epidemiological markers for deduction of mode of contamination. AB - The lysis-centrifugation blood culture system, Isolator, is a promising system with respect to detection of many significant microorganisms, e.g. Staphylococcus aureus and Enterobacteriaceae, as compared with conventional systems. A drawback of the Isolator system is a disturbingly high rate of clinically insignificant, supposedly contaminating coagulase-negative staphylococci, which leads to considerable waste of time and materials in the laboratory. Several sources of these isolates have been proposed (viz. the patient, the ward environment, the laboratory handling, and the plate media). The aim of this study was to pinpoint the origin of these clinically doubtful coagulase-negative staphylococci, using different epidemiological markers, such as species identification, antibiotic susceptibility patterns, phage-types, and plasmid profiles. Plasmid profile analysis proved to be more discriminating than the other techniques and made it possible to conclude that the laboratory handling of the Isolator system was a major source of coagulase-negative staphylococci in this system. PMID- 3044399 TI - Studies of the MHC class I molecules of veto-active and veto-inactive lectin stimulated spleen cell populations. AB - Mouse spleen cells activated for three days with ConA, but not with PHA and LPS, were found to possess strong veto activity. Thus, ConA-activated spleen cells were capable of preventing a cytotoxic T cell response generated in MLC against the H-2 haplotype of the ConA-activated cells. Radiolabelling and chemical cross linking of veto-active and veto-inactive cells, followed by immunoprecipitation of H-2, class I antigens, and analyses of the precipitates by one and two dimensional SDS PAGE were performed. By these procedures we were unable to demonstrate qualitative and quantitative differences between precipitates of veto active and veto-inactive cell populations. PMID- 3044400 TI - Non-neoplastic pulmonary multinucleate giant cells. Comparative study of multinucleate giant cells in porcine dermatosis vegetans and other polykaryocytic conditions. AB - Pulmonary multinucleate giant cells (MGCs) in porcine dermatosis vegetans (DV) were compared with virus-induced and inflammatory-associated MGCs, with particular attention to mechanisms involved in the formation of such cells. Ultrastructurally, pigs with DV showed numerous pulmonary MGCs with transversely sectioned interdigitating microvillous structures. Microvilli were observed closely apposed to type-II pneumocytes and with morphological characteristics of membrane bridge formation. Rabbits infected with Mycobacterium bovis exhibited multiple pulmonary granulomas with prominent MGC formation 14 days after infection. Close apposition between MGCs and small macrophages and membrane bridge formation were demonstrated ultrastructurally. Pulmonary changes in calves infected with bovine respiratory syncytial virus (BRSV) included lobular consolidation and intense infiltration of inflammatory cells into the bronchial, bronchiolar and alveolar walls. Proliferation of bronchiolo-alveolar epithelial cells was prominent, but syncytiae were only occasionally demonstrated. This study of the morphological characteristics of MGCs is related to the present knowledge of mechanisms involved in membrane fusion and allows for an interpretation of an active fusion process involved in the formation of MGCs in porcine DV. PMID- 3044401 TI - Enzyme-linked immunosorbent assay compared with indirect immunofluorescence test for detection of Pneumocystis carinii specific immunoglobulins G, M, and A. AB - An ELISA to measure Pneumocystis carinii-specific IgG, IgM, and IgA has been developed. The antigen was prepared from purified cysts by sonication and ultracentrifugation. Antigen particles with sedimentation coefficients less than 165 S were used. The technique has been compared with indirect immunofluorescence, using whole cysts as antigen. Ninety human sera were assayed. The results were significantly correlated. The ELISA-technique was more sensitive, and owing to the soluble antigen the daily variation was less than 1 per cent. The technique is useful for quick and reliable detection of Pneumocystis carinii antibodies in a routine laboratory. PMID- 3044402 TI - Clonal distribution of septicaemic Escherichia coli O78 isolates, evidenced by enzyme electrophoretic polymorphism. PMID- 3044403 TI - Comparison of bone formed intramuscularly after transplantation of scapular and calvarial osteoblasts. AB - Previous work suggested that osteoblasts determine the size of the bone marrow area within the bone and that calvarial osteoblasts differ from those induced intramuscularly by cartilage formed by transplanted epiphyseal chondrocytes. This study reports morphological observations of bone formed by transplanted scapular and calvarial osteoblasts isolated from bones of young rats. In intact scapulas of 28-day-old rats the percentage area occupied by bone tissue in relation to bone marrow was 6 times larger than in parietal bones of comparable age. Isolated syngeneic scapular osteoblasts usually produced an ossicle with similar general structure and ratio bone tissue/bone marrow area as in intact scapulas. In transplants of calvarial osteoblasts numerous islands of bone tissue with a small amount of bone marrow appeared. Bone formed in allogenic transplants was rejected. These results suggest that osteoblasts from endochondral scapular bone may have different properties than those from intramembranous calvarial bones. Alternatively, the large amount of medullary space in bone produced by transplanted scapular osteoblasts could result from their contamination with bone marrow stromal cells. PMID- 3044404 TI - Forty years of bone cell biology research at the University of Leiden. Dedicated to P.J. Gaillard for the 40th anniversary of the Laboratory of Cell Biology and Histology, University of Leiden, The Netherlands. PMID- 3044405 TI - Osteoclastic bone resorption: in vitro analysis of the rate of resorption and migration of individual osteoclasts. AB - Osteoclasts isolated from the long bones of newborn rabbits were cultured on translucent devitalized bone slices and observed by phase-contrast time-lapse cinemicrography and scanning electron microscopy (SEM). This has allowed us to measure the rate of resorption and the rate of migration of individual osteoclasts. Our films show that osteoclasts do not resorb while migrating. When the osteoclastic resorption areas, which are easily recognizable with phase contrast microscopy as areas delineated by refractile lines, were observed by SEM, such areas appeared as excavated areas lined with a network of collagen fibrils. The rate of migration was calculated using time lapse recordings, and varied from 30 micron/hr to 248 micron/hr, with a mean +/- SEM of 105 +/- 10 micron/hr. The rate of resorption by individual osteoclasts was calculated using both time lapse and SEM data, and varied from 43 micron 3/hr to 1225 micron 3/hr with a mean +/- SEM of 390 +/- 109 micron 3/hr. Additional observations indicated not only that the same osteoclast can resorb at a different rate at different times without any definable alteration of the culture conditions, but also that the same osteoclast can simultaneously resorb two lacunae at different rates. These observations provide, for the first time, data on the rate of resorption and the rate of migration of individual osteoclasts on a bone substratum. PMID- 3044406 TI - Role of mineralizing cartilage in osteoclast and osteoblast recruitment. AB - Periost-free, live and/or devitalized cartilaginous long bone rudiments of fetal mice were transplanted under the renal capsule of adult syngeneic mice to study the role of cells and intercellular matrix in the recruitment and formation of osteoclasts and osteoblasts, both identified by means of enzyme- and immunohistochemical methods. Live bone rudiments recruited host-derived osteoclasts within 5 days after transplantation. Osteoblasts developed as rapidly as osteoclasts and participated in the modeling of the rudiments into hemopoietic bone marrow containing ossicles. Devitalized bone rudiments, killed before osteoclastic invasion had occurred, did not recruit osteoclasts or osteoblasts, and were not resorbed up till 35 days after transplantation. Co-transplantation of live and devitalized bone rudiments however resulted in osteoclastic resorption of the killed rudiments, starting 9 days after transplantation. Again the live rudiments were modeled into ossicles. Devitalized bone rudiments which had been invaded by osteoclasts before killing and transplantation, did recruit host osteoclasts, but at a slower rate than live rudiments, and depending on the number of resorption sites at the time of transplantation. Osteoblasts were not formed. These data suggest that in developing long bones chondrocyte activity is involved in the recruitment of osteoclasts as well as osteoblasts. Matrix components diffusing from resorbing surfaces seem to be involved in osteoclast recruitment. PMID- 3044407 TI - HRH the Princess of Wales, Hon FDSRCS Eng. PMID- 3044408 TI - Badges of the dental profession. BDA Wolverhampton Branch. PMID- 3044409 TI - Internal diameter of the common femoral artery in patients with insulin-dependent diabetes mellitus. AB - By means of an ultrasonographic technique the systolic and diastolic diameters of the common femoral artery were investigated in a group of 50 young insulin dependent diabetics selected as being free from late diabetic complications and atherosclerotic involvement. After correction for normal physiologic variations there was no correlation of arterial diameters to duration of diabetes and no statistical difference in relation to a control group. It is known that there is a progressive increase of the arterial wall thickness in medium-sized arteries in diabetes. Therefore, it is concluded that there is a corresponding dilatation of the arteries. The reasons for this dilatation are discussed from a biophysical point of view. Furthermore, it is concluded that the increased arterial wall stiffness caused by an increasing elastic modulus and thickness reflects the earliest changes in the diabetic macroangiopathy. Occlusions and narrowing seem only to exist in patients with severe late diabetic complications. PMID- 3044410 TI - Asymptomatic abdominal hydatidosis detected by ultrasonography. AB - Ultrasonography (US) and serology (double diffusion for antigen 5 (dd5] were used for the screening of hydatid disease in 1,418 asymptomatic individuals from two provinces in Argentina. A total of 122 positive subjects were identified and in 120 of them lesions consistent with hydatid disease were observed at US (98.4%) while only 19 had positive dd5 tests (15.8%). Chest radiography was performed in 647 of the subjects and an additional 4 lesions were found. Surgery was performed in 32 patients, confirming in all of them the sonographic diagnosis. It is suggested that US should be used in the screening for hydatidosis in high-risk populations as well as in follow-up programmes. PMID- 3044411 TI - Ultrasonography of the hand in rheumatoid arthritis. AB - High resolution ultrasonography of the hand and wrist was performed on 20 patients with definite or probable rheumatoid arthritis (ARA standard criteria) in its early stage. In all the patients, swelling of the soft tissues of the fingers corresponded to an enlargement of the joint capsule containing a hypoechoic exudate. The rheumatoid nodules appeared as fluid-filled rounded cavities with sharp borders. Rheumatoid tenosynovitis was observed in 18/20 patients. This corresponded to oval or spindle-shaped cavities with a hypoechoic (10/18 cases) or anechoic content (8/18 cases) and with the tendon ribbon inside. Rupture of a tendon was diagnosed in 8/20 cases and it was always confirmed at surgery. Tenosynovitis of the flexor carpi ulnaris at the wrist level was observed in 10/20 patients. Ultrasonography is proposed as an effective first line approach and as a periodical follow-up survey in early stage rheumatoid arthritis, in combination with standard radiography. PMID- 3044412 TI - Oesophageal echocardiography. PMID- 3044413 TI - Doppler assessment of the interventricular pressure drop in patients with ventricular septal defects. AB - Doppler ultrasound was used to assess the pressure drop between the ventricles in 109 infants and children (61 less than two years old) with a ventricular septal defect who underwent cardiac catheterisation. The pressure in both ventricles was measured at catheterisation in 103 patients either simultaneously through two catheters (41) or with a single catheter withdrawn across the septum or removed from one ventricle to the other (62). When pressure was measured simultaneously with two catheters (41 patients) the peak to peak and instantaneous gradients showed a maximum difference of 20 mm Hg with levels within 10 mm Hg of each other in 36. Comparison of the difference in the gradients with the average of the measurements demonstrated a tendency for Doppler to underestimate the difference when it was high (greater than 50 mm Hg) and overestimate it when it was low. A Doppler estimate of a low pressure difference between the ventricles indicates pulmonary arterial hypertension and a high one low pulmonary artery pressure, but in the intermediate group Doppler is as yet not sufficiently sensitive to allow selection of those patients who require further investigation and possible operation. Doppler ultrasound was found to be a sensitive method of detecting a very small ventricular septal defect. Thus although Doppler is a very useful means of assessing and following patients with a ventricular septal defect, further studies are required to determine its exact place in clinical practice. PMID- 3044415 TI - Coronary artery-right ventricular fistula after endomyocardial biopsy. AB - A coronary artery-right ventricular fistula developed after endomyocardial biopsy in a recipient of an orthotopic cardiac transplant. PMID- 3044416 TI - Perioperative myocardial infarction. PMID- 3044414 TI - Neonatal anatomical correction of transposition of the great arteries: non invasive assessment of haemodynamic function up to four years after operation. AB - Intracardiac and great artery blood flow velocities were recorded by pulsed and continuous wave Doppler ultrasound in 18 children aged between eight months and four years (mean 25 months) who had undergone anatomical correction of transposition of the great arteries in the first month of life. Postoperative peak flow velocities across the mitral valve and in the ascending aorta were not significantly different from those in an age matched control population, but tricuspid flow velocities were higher than normal. Aortic regurgitation was detected in only one of the eighteen patients, a markedly lower frequency than that reported after two stage anatomical correction. Peak velocities in the pulmonary artery were higher than normal, and in most cases there was some degree of stenosis of the pulmonary artery at the site of anastomosis. PMID- 3044417 TI - Aetiology of ischaemic heart disease. AB - There is evidence from population studies of strong positive associations between smoking, arterial pressure and serum cholesterol concentration and risk of ischaemic heart disease, and these appear to be independent of one another, but the evidence from unifactorial trials of risk factor reduction is less impressive. For smoking, there is no good experimental evidence that cessation reduces the risk of IHD, but there are good observational data to show that those who choose to cease have a lower mortality from IHD, and all causes of death, compared with those who continue to smoke. For arterial pressure, none of the randomized controlled trials has shown any benefit from a reduction in arterial pressure in relation to IHD, although the same studies have shown that risk of stroke is reduced substantially. The best trial evidence for risk factor reduction is seen for serum cholesterol, and in all three major placebo control trials, reduction was associated with a corresponding decrease in the incidence of IHD; for each 1% decrease in serum cholesterol concentration there was a 3% reduction in risk of IHD. However, total mortality was unchanged, and in the case of the WHO Co-Operative Study there was a statistically significant excess of deaths in the treatment group. The use of drugs in the primary prevention of IHD is a matter of judgement based on the risk of disease, the efficacy of the treatment and side effects. PMID- 3044418 TI - Assessment of the patient with known or suspected ischaemic heart disease for non cardiac surgery. AB - The perioperative care of the patient with known or suspected ischaemic heart disease requires careful co-ordination among the anaesthetist, surgeon and medical consultant. Careful preoperative evaluation can aid the identification of risk, the selection of preoperative interventions that may reduce risk, and decisions on intraoperative and postoperative monitoring. Although uncontrolled observational data suggest that such careful management allows for improved outcome, more carefully controlled studies are required to determine the precise benefits and cost-effectiveness of many of the expensive and often invasive interventions that are currently available. PMID- 3044420 TI - Anaesthesia and ischaemic heart disease: laboratory results. PMID- 3044419 TI - Experimental models of myocardial ischaemia. PMID- 3044421 TI - Myocardial ischaemia associated with general anaesthesia. A review of clinical studies. PMID- 3044422 TI - Anaesthetic considerations for the patient with coronary artery disease. PMID- 3044423 TI - Anaesthesia for coronary artery bypass graft. PMID- 3044424 TI - Effect of bile on cyclosporin absorption in liver transplant patients. AB - 1. We quantitated the effect of biliary diversion on cyclosporin (CyA) absorption in liver transplant patients. Multiple blood samples were obtained over one dosing interval following oral CyA administration in eight liver transplant patients before and after T-tube clamping. 2. Cyclosporin concentrations were measured by a high pressure liquid chromatographic method. 3. The mean (+/- s.d.) dose-normalized area under the blood concentration vs time curve (AUC) over a dosing interval was 5.23 (+/- 3.22) ng ml-1 h during the pre clamping period and 15.79 +/- 7.92 ng ml-1 h during the post clamping period. A significant (P less than 0.05) increase (276%) in the dose normalized AUC was observed following the T-tube clamping. 4. Analysis of bile revealed less than 1 mg (less than 1% of dose) of unchanged CyA to be eliminated in bile over 12 h. Therefore the increased CyA AUC/dose following T-tube ligation cannot be explained by enterohepatic recirculation. 5. Increased bile flow secondary to clamping of the T-tube most likely increased the absorption of CyA. Increase in CyA absorption may be due to a bile mediated increase in CyA solubility or gastrointestinal membrane permeability, or increased residence time at the site of absorption. 6. Independent of the mechanism involved, our results indicate that adjustments in CyA dosage must be made whenever external bile diversion is instituted or discontinued. PMID- 3044425 TI - Effects of nifedipine, verapamil and diltiazem on renal function. AB - Eight healthy female volunteers were given single doses of nifedipine 10 mg, verapamil 80 mg, diltiazem 120 mg or placebo on 4 different study days. Urine and sodium output increased after all four drugs, but the increase after nifedipine was significantly greater than after verapamil or placebo. Nifedipine caused a significant increase in heart rate; none of the drugs caused any change in blood pressure. Potassium excretion, creatinine clearance, free water clearance and plasma renin activity did not change with any drug. The mechanism of the natriuresis and diuresis has still to be elucidated. PMID- 3044426 TI - Factors affecting the clinical response to treatment with digoxin and two calcium antagonists in patients with atrial fibrillation. AB - It has been suggested that patients in whom atrial fibrillation (AF) is associated with poor exercise tolerance respond better to treatment with xamoterol plus digoxin than to digoxin alone; this may be attributable to better control of exercise induced tachycardia. We have examined data obtained during studies comparing digoxin and two calcium antagonists in the treatment of AF to see whether subgroups of patients with particularly poor exercise tolerance, rheumatic heart disease or rapid post-exercise heart rates might derive particular benefit from one modality of treatment as opposed to another. The results do not indicate that calcium antagonists improve exercise tolerance compared with digoxin in any of these subgroups despite achieving consistently better control of exercise induced tachycardia. PMID- 3044427 TI - Warfarin dosage requirements: prospective clinical trial of a method for prediction from the response to a single dose. AB - We have previously described a model for predicting individual daily maintenance dosage (MD) requirements of warfarin 24 h after the administration of a single dose. This model relies on measurement of the initial anticoagulant response as the 24 h percentage fall in plasma clotting factor VII activity. It permits prediction of the individual MD given the size of the initial dose, a baseline and desired maintenance value of the prothrombin ratio, and a baseline and 24 h plasma level of factor VII activity. We now present the results of a prospective clinical trial of the method. Data from 65 patients were suitable for analysis. The mean daily MD of warfarin was 4.0 mg (range 1-10 mg). There was a moderately strong linear relationship between predicted and actual MDs of warfarin (r = 0.66, P less than 0.001). Actual vs predicted MDs in individual patients were not significantly different. The mean difference was 0.39 mg. The results of this prospective trial suggest that our model predicts warfarin MD requirements with reasonable accuracy. Nevertheless, the accuracy of the model is not sufficient to replace careful clinical and haematological monitoring of each patient commencing warfarin therapy. PMID- 3044428 TI - Adjuvant tamoxifen for early breast cancer. PMID- 3044429 TI - Detection of circulating antibodies against c-myc protein in cancer patient sera. AB - We have partially purified an archaebacterial protein of 84 kD which shares common epitopes with the human c-myc protein as shown by its cross-reactivity with a commercialized anti-human c-myc antiserum. An antiserum raised against the 84 kD protein recognizes a 60 kD protein from HL-60 nuclei. This protein is also recognized by the anti-human c-myc antiserum. Using this archaebacterial protein as antigen for Western blot analysis, we found that the human c-myc oncogene product could be immunogenic and that it is possible, in some spontaneously occurring human tumours, to detect antibodies against the c-myc gene product in the serum of cancer patients. PMID- 3044430 TI - An in vitro study comparing the cytotoxicity of three platinum complexes with regard to the effect of thiol depletion. AB - The cytotoxicity of three platinum complexes, cis-diamminedichloroplatinum(II) (cis-platin), cis-dichloro-trans-dihydroxy-cis-bis (isopropylamine) platinum(IV), (CHIP) and diammine (1, 1-cyclobutane-dicarboxylato) platinum(II) (carboplatin) on Chinese Hamster ovary (CHO) and mouse sarcoma RIF-1 cells cultured in vitro has been compared. The tumour cell line was much more sensitive to the cytotoxic action of the three agents compared to the CHO cell line. CHIP and carboplatin gave similar dose-response curves, both being much less toxic than cis-platin. The effect of thiol modification on platinum toxicity was also investigated. Substantial reduction in the intracellular non-protein sulphydryl content markedly enhanced the cytotoxicity of CHIP but had much less effect on carboplatin and cis-platin. Thiol depletion by diethylmaleate had a negligible effect on cis-platin toxicity. PMID- 3044432 TI - The Christie hospital adjuvant tamoxifen trial--status at 10 years. AB - From November 1976 to June 1982, a randomised clinical trial was carried out at the Christie Hospital, Manchester, to test the clinical efficacy of tamoxifen (TAM) as an adjuvant to surgery for patients with operable breast carcinoma. Following surgery, premenopausal women were randomly allocated to have either TAM 20 mg day-1 for one year or an irradiation menopause (the previous standard treatment). Postmenopausal women had TAM 20 mg day-1 for one year or no further treatment (Controls). A total of 1005 patients were entered into the trial of whom 961 are evaluable at 10 years from the inception. At 10 years the analysis shows no significant difference in overall and disease free survival between premenopausal women given TAM or an irradiation menopause. For premenopausal node negative patients there would appear to be a trend in favour of the TAM treated patients with a 93% ten year survival vs. 82% for the irradiation menopause group (P = 0.09). When the disease free survival of all 961 patients is analysed, allowing for node status, then there is a marked trend in favour of the TAM treated patients (P = 0.07). Of the patients originally allocated to TAM 47% had an irradiation menopause on relapse and 73% of the postmenopausal control patients had TAM on relapse. The incidence of side effects and second primary tumours is discussed as well as the possible effects of varying the length of time over which adjuvant TAM is administered. PMID- 3044433 TI - The relationship between histological grade, oestrogen receptor status, events and survival at 8 years in the NATO ('Nolvadex') trial. AB - A pathological review was carried out on 600 patients with breast carcinoma entered into the 'Nolvadex' Adjuvant Trial Organisation (NATO) study. The tumours were graded histologically and these results were compared with the oestrogen receptor (ER) status of the tumours, the numbers of recurrences and the length of survival of the patients. It was found that histological grading was predictive both in terms of events and survival, and correlates significantly with oestrogen receptor status; within histological grades I and II, patients receiving 'Nolvadex' had fewer events and deaths compared with patients in the control group. For patients with grade III tumours qualitatively it was in the same direction as the benefit obtained in patients with grade I and II tumours. PMID- 3044431 TI - Transforming growth factor-beta: possible roles in carcinogenesis. AB - TGF-beta is the prototype of a large family of multifunctional regulatory proteins. The principal sources of the peptide, platelets and bone, suggest that it plays a role in healing and remodeling processes. In vitro, TGF-beta is chemotactic for monocytes and fibroblasts and can greatly enhance accumulation of extracellular matrix components by fibroblasts. Its ability to stimulate the formation of granulation tissue locally and the demonstration of specific time- and tissue-dependent expression in embryogenesis suggest that similar mechanisms are operative in vivo. By analogy to its effects in wound healing and embryogenesis, it is proposed that TGF-beta, secreted by tumour cells, can augment tumour growth indirectly by effects on the stromal elements. These effects include suppression of the immune response, and enhancement of both angiogenesis and formation of connective tissue. Many tumour cells have escaped from direct growth inhibitory effects of TGF-beta by a variety of mechanisms including inability to activate the latent form of the peptide, loss of cellular receptors for TGF-beta, and loss of functional intracellular signal transduction pathways. PMID- 3044434 TI - Does chemotherapy improve survival in advanced breast cancer? A statistical overview. AB - The relative efficacies of cytotoxic chemotherapy regimens in the treatment of advanced breast cancer are generally assessed by comparing response rates in randomised trials. Treatment attempts to prolong survival but trials rarely demonstrate a statistically significant survival advantage: it has been argued that chemotherapy does not prolong survival. The correlation between response rates and survival has been examined by reviewing 79 comparisons between arms with unequal response rates in 50 published trials of chemotherapy in advanced breast cancer. In 73% of comparisons the group with the higher response rate also demonstrated the longer median survival (P less than 0.001). Weighted linear regression showed a statistically significant relationship between relative response rates and survival (P less than 0.001). The number of patients in a comparison did not influence this relationship. PMID- 3044435 TI - Plasma insulin response to oral glycaemic stimulus in lichen planus. PMID- 3044436 TI - Annotation. Myelofibrosis: prognostic factors and treatment. PMID- 3044437 TI - Confirmation of bone marrow engraftment by detection of M/N blood group antigens on erythroblasts in erythroid bursts. AB - In order to confirm erythroid engraftment, we examined the changes in blood group antigens expressed on erythroblasts in the haematopoietic colonies of a bone marrow transplant recipient. A 44-year-old female with acute myelogenous leukaemia underwent an allogeneic bone marrow transplantation from her sister donor. Prior to the transplantation, the blood groups of both recipient and donor were analysed, and their M/N groups found to differ, the former being MN and the latter N. Bone marrow mononuclear cells were obtained from the patient on day 21 after bone marrow transplantation and cultured in semi-solid medium. Erythroblasts were collected from the erythroid bursts that had formed, and were subjected to flow cytometry using monoclonal anti-M and anti-N. Anti-N reactive cells accounted for 99.2% of those in the erythroid bursts, while only 0.3% of these cells were anti-M reactive. MN type erythrocytes in the recipient's peripheral blood had been replaced by N type and only 2.0% of erythrocytes were anti-M reactive on the 70th day after BMT. These erythroblasts and erythrocytes were phenotypically N, and originated from the donor haemopoietic stem cells with the success of bone marrow engraftment. From our findings, it appears that the M/N blood group antigens were produced by the erythroid cells themselves. PMID- 3044438 TI - Marrow transplantation from HLA non-identical family donors for the treatment of leukaemia: a pilot study of 15 patients using additional immunosuppression and T cell depletion. AB - Fifteen patients with high-risk leukaemia were given T-cell depleted marrow transplants from HLA non-identical related donors. They were treated with a combination of total body irradiation (TBI), high-dose cytosine arabinoside (Ara C) and high-dose melphalan in an attempt to prevent a host-versus-graft reaction. Antilymphocyte globulins were given prior to transplantation for additional immunosuppression to 13 patients and in-vivo monoclonal antibody anti-human LFA1 to two. Engraftment and chimaerism assessed by HLA typing were achieved in 14 patients. Seven developed acute graft-versus-host disease (two fatal), one failed to engraft. Six patients died in complete remission from cytomegalovirus (CMV) interstitial pneumonitis and three remain alive in complete remission 2, 3 and 13 months after transplant. We conclude that aggressive immunosuppression allows for sustained engraftment of T-cell depleted HLA non-identical marrow. The incidence and severity of GVHD are acceptable and CMV pneumonitis remains the major problem. PMID- 3044439 TI - Allogeneic bone marrow transplantation for acute leukaemia: comparative outcomes for adults and children. AB - Advances in both allogeneic marrow transplantation and conventional therapy for acute leukaemia have complicated the choice between bone marrow transplantation (BMT) and other remission treatment options. Because older patients may be more susceptible to BMT-related complications, this study analysed the effect of age on clinical outcome for 149 patients with acute leukaemia in remission receiving allogeneic BMT. Overall projected relapse-free survival at 3 years post transplant is equivalent for 48 adults (18 years or older) and 101 children (less than 18) at 45.4% (31.1-59.6; 95% confidence interval) and 39.9% (30.1-49.7; 95% C.I.), respectively. Among 73 patients with acute lymphocytic leukaemia (ALL) 35.3% of adults and 30.1% of children survive relapse-free at 3 years. Cox multiple regression analysis demonstrated that higher diagnostic white count, but not pre-transplant extramedullary leukaemia, remission number, or age, was important as an independent adverse clinical prognostic factor for patients with ALL. Overall outcome was better for 76 patients with acute myeloid leukaemia (AML) with 51.6% of adults and 52.9% of children surviving relapse-free at 3 years post-transplant. Cox multivariate regression analysis identified first remission status and lower white cell count, but not patient age, as independent predictors of improved relapse-free survival for AML patients. Adults had greater transplant morbidity, predominantly related to a higher incidence of acute graft versus-host disease (GVHD), resulting in longer hospital stay. Survival at 100 d, long-term survival and clinical performance status were similar in both age groups. These data suggest that results of allogeneic BMT for adults with acute leukaemia compare favourably with those found in children and are superior to most reports of conventional chemotherapy. Allogeneic BMT remains a reasonable option for remission acute leukaemia patients up to the age of 45. PMID- 3044440 TI - Late haematological complications in severe aplastic anaemia. AB - 137 patients with severe aplastic anaemia (SAA) were treated in Basel from 1976 to 1986. 34 underwent bone marrow transplantation (BMT) and 103 received antilymphocyte globulin (ALG) therapy. We have analysed the incidence of late haematological complications in both groups of patients. 20 patients treated with ALG developed a late haematological complication. A myelodysplastic syndrome or frank leukaemia occurred in eight and paroxysmal nocturnal haemoglobinuria (PNH) in 13 patients. Nine of the 13 patients with PNH had clinical signs of haemolysis, four only had positive laboratory tests. One patient had PNH and acute leukaemia. The risk of developing a haematological complication increased continuously and reached 57% at 8 years. Neither PNH nor leukaemia occurred in patients treated with BMT. The increased survival rate and the long observation time after ALG therapy have revealed a new perspective of the prognosis of aplastic anaemia. Patients treated with BMT appear to be cured whereas those treated with ALG remain at risk for late complications. PMID- 3044441 TI - Competition between recipient and donor cells after bone marrow transplantation for chronic myeloid leukaemia. AB - The cytogenetic and clinical course of three patients allografted for Ph positive chronic myeloid leukaemia are reported. All patients had a peculiar pattern of relapse. Two out of three patients had donor marrow graft pretreated with monoclonal antibody for graft versus host prevention. The cytogenetic relapse was invariably associated with major morphological changes in the marrow indicating that these were also haematological relapses. However, no changes in the peripheral blood count were observed. When relapse occurred in these patients, Ph positive marrow metaphases and host red blood cells ranged from 75% to 100% of the total cell population: thereafter they spontaneously reverted to complete chimaerism. Therefore the presence of leukaemic cells even in considerable amount was not sufficient, per se, to prevail over normal marrow. In addition these observations indicate that relapse was not associated with elimination of the graft: while haemopoiesis was entirely of recipient origin the donor normal stem cells were present and vital although functionally silent. These data suggest that, although TBI remains the more effective tool for eradicating the majority of leukaemic cells, haemopoietic competition between host and donor marrow may have a major impact on leukaemic relapse. PMID- 3044442 TI - Determination of the growth fraction in monoclonal gammopathy with the monoclonal antibody Ki-67. AB - The monoclonal antibody Ki-67 reacts with a nuclear antigen that is present only in proliferating cells. The proportion of Ki-67 positive cells may therefore serve as a reliable measurement for the growth fraction in normal and neoplasmic cell populations. We have tested the significance of the MoAb Ki-67 in the classification of monoclonal gammopathy and compared the results with the plasma cell labelling index. In benign monoclonal gammopathy the percentage of Ki-67 positive plasma cells (median 1.6%) was significantly lower than in untreated multiple myeloma (median 9.6). Among the patients with more than 10% Ki-67 positive plasma cells there were some very short survivors. The largest growth fractions (median 41.8%) were found in patients with relapsing multiple myeloma indicating here a different growth pattern more resembling the high-grade lymphomas. A linear correlation between the proportion of Ki-67 positive plasma cells and the labelling index was not found. Determination of the plasma cell growth fraction with the monoclonal antibody Ki-67 in monoclonal gammopathy may help to discriminate benign monoclonal gammopathy from multiple myeloma and will probably identify a subgroup of multiple myeloma patients with a poor prognosis, including those with relapsing multiple myeloma. PMID- 3044443 TI - Amino acid absorption and production of pancreatic hormones in non-anaesthetized pigs after duodenal infusions of a milk enzymic hydrolysate or of free amino acids. AB - 1. Six non-anaesthetized pigs (mean body-weight 57.0 kg) were used to study the intestinal absorption of amino acids (AA) from either an enzymic hydrolysate of milk (PEP) containing a large percentage of small peptides (about 50% with less than five AA residues) and very few free AA (8%), or from a mixture of free AA with an identical pattern (AAL) infused intraduodenally in one of two amounts (55 or 110 g). Concomitant insulin and glucagon production rates were estimated. 2. Each pig was previously fitted, under anaesthesia, with an electromagnetic flow probe around the portal vein, with permanent catheters in the portal vein, the carotid artery and the duodenum. Each infusion was performed after an 18 h fasting period and each pig received each infusion. The observation period lasted for 5 h. 3. The absorption of AA was greater, more rapid and more homogeneous after PEP infusion than after AAL infusion, independent of the amount infused. 4. For the majority of AA considered individually, the absorption coefficient was higher after infusion of PEP than after that of AAL. The exceptions were methionine with a higher absorption coefficient after AAL infusion, and isoleucine, aspartic acid + asparagine and glutamic acid + glutamine with identical coefficients for both infusions. 5. Some AA, such as asparagine, ornithine, citrulline and taurine, while absent in the infusates, appeared in the portal vein in appreciable amounts after the infusion of both solutions. While a small proportion of taurine may arise from recycling of taurine-containing bile salts, it seems that the gut wall is able to synthesize all four AA. 6. Insulin production did not differ according to the nature or amount of solutions infused. Glucagon production was greater after PEP infusion. PMID- 3044445 TI - Morphoregulatory molecules. PMID- 3044444 TI - The effect of underfeeding on the physiological response to food ingestion in normal weight women. AB - 1. The thermogenic, cardiovascular and metabolic responses to the ingestion of a 30 kJ/kg body-weight test meal were studied in six normal weight, female subjects before and after a 7 d period of underfeeding at 60 kJ/kg ideal body-weight per d. 2. With underfeeding there were decreases in body-weight, plasma insulin and 3,5,3'-triiodothyronine concentrations, resting metabolic rate and respiratory exchange ratio, with increased blood ketone levels. Baseline 'arterialized' venous plasma noradrenaline and adrenaline concentrations were not affected by underfeeding. 3. Ingestion of the test meal caused similar increases in heart rate and calf blood flow and changes in blood pressure in the fed and underfed states. There was a greater glycaemic response to the test meal in the underfed state compared with the fed state although the rise in plasma insulin concentration was similar and ketogenesis was suppressed. The increases in metabolic rate and plasma noradrenaline concentrations following the test meal were similar in the fed and underfed states. 4. Although the period of underfeeding in the present study led to considerable metabolic adaptation, and some alteration in physiological responses to ingestion of a test meal, there was no evidence that there were associated changes in sympathetic nervous system activation. PMID- 3044446 TI - Dinitrogenase with altered substrate specificity results from the use of homocitrate analogues for in vitro synthesis of the iron-molybdenum cofactor. AB - The in vitro synthesis of the iron-molybdenum cofactor (FeMo-co) of nitrogenase requires homocitrate (2-hydroxy-1,2,4-butanetricarboxylic acid). Homocitrate is apparently synthesized by the nifV gene product. In the absence of homocitrate, no FeMo-co is formed in vitro, as determined from coupled C2H2 reduction assays and the lack of 99Mo label incorporation into apodinitrogenase. Several organic acids were tested for their ability to replace homocitrate in the FeMo-co synthesis system. With appropriate homocitrate analogues, aberrant forms of FeMo co are synthesized that exhibit altered substrate specificity and inhibitor susceptibility. Homoisocitrate (1-hydroxy-1,2,4-butanetricarboxylic acid) and 2 oxoglutarate facilitated the incorporation of 99Mo into apodinitrogenase in the FeMo-co synthesis system, yielding a dinitrogenase that effectively catalyzed the reduction of protons but not C2H2 or N2. Citrate also promoted the incorporation of 99Mo into apodinitrogenase, and the resulting holodinitrogenase reduced protons and C2H2 effectively but not N2. In addition, proton reduction from this enzyme was inhibited by CO. The properties of the homodinitrogenase formed in the presence of citrate were reminiscent of those of the Klebsiella pneumoniae NifV- dinitrogenase. We also observed low rates of HD formation from NifV- dinitrogenase compared to those from the wild-type enzyme. No HD formation was observed with the dinitrogenase activated in vitro in the presence of citrate. We propose that in vivo NifV- mutants utilize citrate for FeMo-co synthesis. PMID- 3044447 TI - Expression and site-directed mutagenesis of human dihydrofolate reductase. AB - A procaryotic high-level expression vector for human dihydrofolate reductase has been constructed and the protein characterized as a first step toward structure function studies of this enzyme. A vector bearing the tac promoter, four synthetic oligodeoxynucleotides, and a restriction fragment from the dihydrofolate reductase cDNA were ligated in a manner which optimized the transcriptional and translational frequency of the enzyme mRNA. The reductase, comprising ca. 17% of the total soluble protein in the host bacteria, was purified to apparent homogeneity as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis and characterized by amino acid composition, partial amino acid sequence, and steady-state kinetic analysis. This expression vector has been used as a template for double-stranded plasmid DNA site-specific mutagenesis. Functional studies on a Cys-6----Ser-6 mutant enzyme support the contention that Cys-6 is obligatory for organomercurial activation of human dihydrofolate reductase. The Ser-6 mutant enzyme was not activated to any extent following a 24-h incubation with p-(hydroxymercuri)benzoate and nicotinamide adenine dinucleotide phosphate (reduced) (NADPH), whereas the kcat for Cys-6 reductase increased 2-fold under identical conditions. The specific activities of the Cys-6 and Ser-6 enzymes were virtually identical as determined by methotrexate titration as were the Km values for both dihydrofolate and NADPH. The Ser-6 mutant showed a decreased temperature stability and was more sensitive to inactivation by alpha-chymotrypsin when compared to the wild-type enzyme. These results suggest that the Ser-6 mutant reductase is conformationally altered relative to the Cys-6 native enzyme. PMID- 3044448 TI - ATP-dependent inactivation and slow binding inhibition of Salmonella typhimurium D-alanine:D-alanine ligase (ADP) by (aminoalkyl)phosphinate and aminophosphonate analogues of D-alanine. AB - In Salmonella typhimurium, D-alanine:D-alanine ligase (ADP) (EC 6.3.2.4) is the second enzyme in the three enzyme D-alanine branch pathway of peptidoglycan biosynthesis. The interaction of this enzyme with a possible transition-state analogue, the (aminoalkyl)phosphinate D-3-[(1-aminoethyl)phosphinyl]-2 heptylpropionic acid [Parsons et al. (1987) Abstracts of Papers, 193rd National Meeting of the American Chemical Society, Denver, CO, MEDI 63, American Chemical Society, Washington, DC], has been studied. This compound is a potent active site directed inhibitor and is competitive with D-alanine (Ki = 1.2 microM); it exhibits time-dependent inhibition in the presence of ATP. Kinetic analysis revealed a rapid onset of steady-state inhibition (kon = 1.35 X 10(4) M-1 s-1) followed by slow dissociation of inhibitory complex(es) with a half-life of 8.2 h. The inhibitory complex was shown to consist of E...I...ATP in equilibrium with E...I, Pi, and ADP. Similar time-dependent inhibition was also observed with D-(1 aminoethyl)phosphonic acid (D-Ala-P) (Ki = 0.5 mM; kon = 27 M-1 s-1; t1/2 for regain = 1.73 min) but not with D-(1-aminoethyl)phosphinic acid, which behaved as a simple competitive inhibitor (Ki = 0.4 mM). The mechanism of inhibition is discussed in the light of the precedents of glutamine synthase inhibition by methionine sulfoximine and phosphinothricin. PMID- 3044449 TI - Two smooth muscle myosin heavy chains differ in their light meromyosin fragment. AB - Smooth muscle myosin heavy chains [SM1, approximately 205 kilodaltons (kDa), and SM2, approximately 200 kDa] were separated on sodium dodecyl sulfate (SDS) polyacrylamide gels. Peptide maps of the two heavy chains showed unique patterns. Limited proteolytic cleavage of purified swine stomach myosin was performed by using a variety of proteases to produce the major myosin fragments which were resolved on SDS gels. A single band was obtained for heavy meromyosin in the soluble fraction following chymotrypsin digestion. However, a variable number of bands were observed for light meromyosin fragments in the insoluble fraction after chymotrypsin digestion. Peptide mapping indicated that the two bands observed after short digestion times with chymotrypsin had relative mobility and solubility properties consistent with approximately 100- and 95-kDa light meromyosin (LMM) fragments. These results indicate that the region of difference between SM1 and SM2 lies in the LMM fragment. PMID- 3044450 TI - Interaction of tRNAs and of phosphorothioate-substituted nucleic acids with an organomercurial. Probing the chemical environment of thiolated residues by affinity electrophoresis. AB - The interactions of 4-thiouridine and 5-[(methylamino)methyl]-2-thiouridine in tRNA and of phosphorothioate esters in nucleic acids with an organomercurial have been investigated. For this purpose, an affinity electrophoretic system has been developed in which the mercury derivative has been covalently immobilized in a standard polyacrylamide gel. The retardation of thiolated macromolecules was found to be sensitive to the chemical environment of the sulfur atom, giving characteristic interaction constants dependent on the nature of the modification and its accessibility to binding. The interaction could, in the case of tRNA, be abolished by conventional specific chemical modification of the thiolated bases, as well as by irradiation with 32P-derived beta-emission. Not only has the fractionation of sulfur-modified from unmodified species been attained but a quantitative application of the technique has made it possible to study the binding of mercury and, by competition, that of magnesium in terms of the conformation of tRNA. PMID- 3044451 TI - The irradiated epidermis inhibits newt limb regeneration by preventing blastema growth. A histological study. AB - It has been established that X-ray irradiation localized to a forelimb or entire irradiation of premetamorphic Pleurodeles larvae prevented limb regeneration. Transplantation of non-irradiated skin, dermis or muscle to limb stumps of locally irradiated newts was sufficient to allow a blastema to develop. Transplantation of the same tissues to limb stumps of entirely irradiated newts yielded different results with the different graft types. Skin graft allowed a normal blastema to be established but dermis or muscle grafts did not. In order to define more precisely the role played by the epidermis in the establishment of a blastema, and in the growth of a regenerate, different combinations of limb tissues, either irradiated or not, were carried out at the level of amputated limb stumps. At four different times (8-10 days; 13-15 days; 20-23 days; 30 days or more) after amputation the stumps were examined in histological longitudinal sections to study the first events of regeneration, that is dedifferentiation and growth. Dedifferentiation occurred in both normal and irradiated tissues of mesodermal origin. The healthy mesenchymal cells began dividing and formed a growing blastema only when associated with a non-irradiated epidermis. Healthy mesenchymal cells covered with an irradiated epidermis exhibited a few mitoses after dedifferentiation, but the mitotic figures became rarer and rarer until the animals died. The lack of dense accumulation of blastemal cells in such limb stumps suggested that the healthy epidermis allows the mesenchymal cells to divide actively to constitute a growing blastema. Hence, X-ray irradiation seems to be responsible for the loss of such an epidermal mitogenic influence on the underlying mesenchymal cells. PMID- 3044452 TI - Effect of chronic modification of diet fat and cholesterol during gestation on plasma hormones and hepatic enzyme activities in rat fetus. AB - Diet-induced elevation of cholesterol and corticosterone in pregnant rats had no effect on fetal plasma cholesterol or corticosterone, 3-hydroxy-3-methylglutaryl coenzyme A or hepatic reductase activities. The data suggest that increasing the maternal cholesterol pool has no effect on cholesterol transfer to, or metabolism in, the developing offspring, and that the late gestation fetus can maintain corticosterone levels appropriate to gestational age despite diet-induced elevations in the maternal plasma. PMID- 3044453 TI - Stress-responsive neuromodulators. PMID- 3044454 TI - The luteinizing hormone-releasing hormone (LHRH) agonist, [D-Trp6, des-Gly NH2(10)]-LHRH ethylamide, has no direct effect on spermatogenesis in the rat. AB - Treatment of intact rats with luteinizing hormone-releasing hormone (LHRH) agonists has been shown to produce atrophy of a variable number of testicular seminiferous tubules. These findings raised the question of a possible direct versus indirect action of LHRH agonists on spermatogenesis. To answer this question, we treated hypophysectomized rats with the LHRH agonist [D-Trp6, des Gly-NH2(10)]-LHRH ethylamide, dihydrotestosterone (DHT), or a combination of these two compounds for a period of 1 mo. Treatment of hypophysectomized animals with the LHRH agonist alone had no significant effect on the atrophy of seminiferous tubules found after hypophysectomy. DHT, however, maintained spermatogenesis at 80% of the level seen in intact animals. When DHT and the LHRH agonist were administered in combination, the stimulatory effects of DHT were observed with no significant interference caused by the LHRH agonist. This study shows that an LHRH agonist has no direct effect on the morphology of the seminiferous tubules in the absence of the pituitary gland and strongly suggests that the atrophy observed in the testis after LHRH agonist treatment in intact animals is mediated by the LHRH agonist-induced changes in luteinizing hormone secretion and/or direct action of the peptide on Leydig cells. PMID- 3044456 TI - AIDS: from exact data to beginning of optimism. AB - We have compiled epidemiologic data on AIDS which failed to support systematic screening for HIV viruses and which allow a certain optimism regarding the future of the epidemic: there seems to be, in New York, a decrease in the frequency of affected homosexuals, probably because they have followed advice on hygiene. Hence, the systemic, pleiomorphic, HIV infectious disease which might include, in addition to AIDS (35%), the forms where different organs are directly affected, is developing into a heterosexual disease affecting drug addicts and their sexual partners. The therapeutic paradigm is underlined. It is unacceptable that those drugs which have proven effective against the virus, or immune functions, should not be used in combination, on account of some of them still being at a trial stage, that is, administered individually and compared to a placebo, after randomization. PMID- 3044455 TI - The neurological features of acute HIV infection. AB - This paper describes the few case reports of neurological effects of acute (primary) HIV infection. Following a typical primary illness (fever, sore throat, headache, rash, lymphadenopathy, superficial oral ulcers, conjunctivitis, leukopenia and thrombocytopenia) aseptic meningitis, myelopathy, spinal myoclonus, peripheral or cranial neuropathy, neuralgia and ganglioneuronitis may occur, usually within 3 weeks. Encephalopathy with spontaneous recovery also occurs, usually without other features of acute HIV infection. Diagnosis depends on demonstration of seroconversion which may be delayed by weeks. No therapy is yet available. PMID- 3044457 TI - The electrophysiological properties of the parathyroid cell: results of a study employing Sprague-Dawley rats and a review of the literature. AB - The present work indicates that the parathyroid cell of the Sprague-Dawley rat has a resting transmembrane potential of -30.5 +/- 4.1 mV. In a review of data obtained by earlier studies using other species, it was clear that in general the parathyroid cell transmembrane potential lies between -20 mV and -45 mV, that the transmembrane K+ concentration gradient is the principal factor in determining the magnitude of the membrane potential in the resting state, and that in most species a reduction in the extracellular Ca++ leads to cellular hyperpolarization. Detailed data on cellular input resistance in intact parathyroid tissue is not as yet available; however, indirect evidence exists that direct communication of the type described between other gland cells is present between parathyroid cells, a phenomenon which allows the passage of current to flow between adjacent cells via specialised connecting structures such as gap junctions. PMID- 3044458 TI - Expression of the granulocyte/macrophage colony-stimulating factor gene in leukemic blast cells from patients with acute non-lymphocytic leukemia. AB - Since granulocyte/macrophage colony-stimulating factor (GM-CSF) has been reported to stimulate the proliferation of clonogenic leukemia cells from patients with acute non-lymphocytic leukemia in vitro, the expression of the GM-CSF gene in primary human leukemic blast cells was studied. T-cell-depleted mononuclear cells in freshly drawn peripheral blood from patients with acute non-lymphocytic leukemia (ANLL) were subjected to the study. The expression of the GM-CSF gene was detected by Northern blotting analysis using [32P]GM-CSF as a probe. The GM CSF gene was expressed in two out of five cases examined. In both of the patients who showed GM-CSF expression, remission could not be achieved. PMID- 3044459 TI - Amino acid transport systems of lysosomes: possible substitute utility of a surviving transport system for one congenitally defective or absent. AB - Ways in which other transport systems may compensate for one that is genetically defective are considered. Comparisons of the transport systems of organelles (here the lysosome) with the transport system at the plasma membrane has significant implications for chemotherapy. PMID- 3044460 TI - The influence of starvation and natural refeeding on the rate of triacylglycerol/fatty acid substrate cycling in brown adipose tissue and different white adipose sites of the rat in vivo. The role of insulin and the sympathetic nervous system. AB - Triacylglycerol/fatty acid substrate cycling was measured in vivo in brown adipose tissue (BAT) and white adipose tissue (WAT) of fed, starved and refed rats. Starvation (24h) significantly decreased the rate of cycling in BAT, and refeeding chow diet led to a rapid, 6-fold increase in cycling. Cycling rate in WAT was much lower than in BAT, and was not influenced by fasting or refeeding. Similar rates of cycling were found in epididymal, mesenteric, subcutaneous, and scapular WAT depots. Sympathetic denervation of interscapular BAT abolished the response of the tissue to refeeding, as did acute suppression of insulin secretion. Similarly, rats fasted for 3 days showed no acute increase in the activity of the cycle following refeeding. PMID- 3044463 TI - Abdominal pain, indigestion, anorexia, nausea and vomiting. AB - Non-specific abdominal complaints are a very frequent cause of discomfort. Even if only comparatively few are brought to the attention of the physician, they account for a considerable portion of the reasons for seeking medical care, both in acute and chronic conditions. On the other hand, few drugs are free of the suspicion of causing abdominal complaints, which make up between one-tenth and one-third of reported adverse reactions. A wide variety of possible alternative or concomitant causes makes a clear causative attribution to suspected drugs very difficult. This holds especially true for the ill-defined conditions of indigestion and anorexia. For nausea and vomiting, specific scales have been developed which facilitate differentiation between drugs causing these effects most frequently and most intensively. They have been applied in cytostatic therapy, where this is one of the most frequently encountered problems, but nausea and vomiting can seriously affect compliance in many other treatments. Somatic abdominal pain results in most instances from the irritation of the parietal peritoneum and is usually the effect of a lesion. This may or may not be caused by a drug, but this cause should be the first consideration. Visceral pain may result from functional disturbance of secretory glands or of the muscular coat, from drug action on bowel content or from irritation of the mucosa, all of which are frequently interrelated. Most frequently suspected pharmacological causes are drugs with anticholinergic action, antibiotics, potassium supplements and non-steroidal, anti-inflammatory agents. Drug-induced hyperinsulinism and porphyria are rare cases. Abuse of laxatives should always be considered because of its prevalence. A great number of other untoward drug effects have been described in the literature, but rarely merit first consideration. With the exception of promptly occurring or persistent emesis, gastrointestinal symptoms usually are not pathognomonic for drug effects and are the result of several factors. The usual approach to identifying an adverse drug effect is to delineate the functional or structural disorder, and to associate this diagnosis with possible pharmacodynamic aetiologies. PMID- 3044462 TI - Drug-induced oesophageal lesions. AB - Persisting retrosternal pain of sudden onset is suggestive of a drug-induced oesophageal lesion, particularly if it starts at night. After exclusion of a myocardial infarction, a carefully taken history and oesophagoscopy will rapidly clarify the cause and severity of the injury. Since almost any pill may produce oesophageal lesions, care has to be taken that tablets, capsules and other pills are always taken in an upright position together with a fluid chaser of at least 120 ml. If possible, less harmful liquid preparations of the drugs should be preferred. Lesions in the oesophageal wall and perioesophageal tissue are almost unavoidable side-effects of sclerotherapy of oesophageal varices. The patient and the doctor should be particularly aware of bleeding from oesophageal ulcers during the first week after sclerotherapy. Numerous drugs may weaken or strengthen contractions of the oesophagus and lower oesophageal sphincter. These potentially unwanted motor effects of the drugs have to be kept in mind, especially in patients with pre-existing gastro-oesophageal reflux disease and hypermotility states. PMID- 3044461 TI - Graft versus leukemia in bone marrow transplantation. PMID- 3044464 TI - Drug-induced pancreatitis. AB - We reviewed the English-language literature pertaining to drug-induced pancreatitis and attempted to determine whether the reported association between each drug and pancreatitis was valid. The following drugs seem to cause pancreatitis: azathioprine, chlorothiazide and hydrochlorothiazide, oestrogens, frusemide, 6-mercaptopurine, methyldopa, sulphonamides, sulindac, tetracycline and valproic acid. Less convincing but suggestive evidence exists for colaspase, chlorthalidone, combination cancer chemotherapy, cimetidine, cisplatin, corticosteroids, cytosine arabinoside, diphenoxylate, ethacrynic acid, iatrogenic hypercalcaemia, methandienone, metronidazole, nitrofurantoin, pentamidine, phenformin, piroxicam and procainamide. In general, pancreatitis is a very rare complication of treatment with these drugs. The pathogenesis of the pancreatitis is usually obscure, but is probably mediated by an immune response. Certain drugs such as oestrogens cause hypertriglyceridaemia, which in turn may lead to pancreatitis. PMID- 3044465 TI - Ulcerogenic drugs and upper gastrointestinal bleeding. AB - Aspirin and other NSAIDs are drugs for which the causal association with major gastrointestinal bleeding has not been adequately or conclusively demonstrated, although a certain degree of correlation is very likely. For aspirin ingestion in particular the increased risk is confined to patients taking the drug at heavy and regular dosages (less than 1% of users), and can be reduced further by the use of enteric-coated formulations. For non-aspirin NSAIDs, the relative risk of GI bleeding after repeated and prolonged exposure (in comparison to controls) has been quantified between 1.5 and 2.7, which is a small but significant figure, and it is increased by the age of the patients, by the duration of treatment and by the dose of drug. No consistent causal relationship can be found between major GI bleeding (or other major peptic ulcer complications) and steroids or other 'ulcerogenic' drugs. The therapy of drug-induced (or drug-associated) GI bleeding is probably not different from the usual treatment of upper GI haemorrhage. As far as the treatment of drug-associated gastroduodenal mucosal damage is concerned, it appears that with mucoprotective agents or H2 antagonists the healing rates of peptic ulcers is slower than observed in non-drug-associated disease. Prophylactic treatment with prostaglandins has only been proposed; and prophylactic treatment with H2 antagonists has been disappointing. PMID- 3044466 TI - Malabsorption syndromes. PMID- 3044467 TI - Diarrhoea and constipation. AB - Drug-induced constipation is mostly caused by changes in gut motility, whilst diarrhoea is more frequently caused by an increase in intestinal fluid secretion. In both instances the drug has to reach the enteric nervous system or the enterocyte, either via the blood or from the lumen, in sufficient concentrations to affect the mediators that regulate motility and fluid transport. Diarrhoea and constipation are frequently mentioned as side-effects of drugs, and therapeutic agents for almost all organ systems have been implicated. However, both these side-effects are usually mild or moderate, and rarely necessitate interruption of drug treatment. An exception to this rule is the antibiotic-associated colitis seen in patients treated with antibiotics such as lincomycin or clindamycin; in principle almost all antibiotics may cause this severe and potentially life threatening complication. Other rare forms of severe, drug-induced colitis and diarrhoea result from toxic or anaphylactic reactions against gold preparations, cytostatic agents and sulphonamides. Ischaemic colitis due to vascular complications has been described in some women taking oral contraceptives, and in patients treated with vasopressin or digitalis. PMID- 3044468 TI - Acute and chronic drug-induced hepatitis. AB - Adverse drug reactions may mimic almost any kind of liver disease. Acute hepatitis is often due to the formation of reactive metabolites in the liver. Despite several protective mechanisms (epoxide hydrolases, conjugation with glutathione), this formation may lead to predictable toxic hepatitis after hugh overdoses (e.g. paracetamol), or to idiosyncratic toxic hepatitis after therapeutic doses (e.g. isoniazid). Both genetic factors (e.g. constitutive levels of cytochrome P-450 isoenzymes, or defects in protective mechanisms) and acquired factors (e.g. malnutrition, or chronic intake of alcohol or other microsomal enzyme inducers) may explain the unique susceptibility of some patients. Formation of chemically reactive metabolites may also lead to allergic hepatitis, probably through immunization against plasma membrane protein epitopes modified by the covalent binding of the reactive metabolites. This may be the mechanism for acute hepatitis produced by many drugs (e.g. amineptine, erythromycin derivatives, halothane, imipramine, isaxonine, alpha-methyldopa, tienilic acid, etc.). Genetic defects in several protective mechanisms (e.g. epoxide hydrolase, acetylation) may explain the unique susceptibility of some patients, possibly by increasing exposure to allergenic, metabolite-altered plasma membrane protein epitopes. Like toxic idiosyncratic hepatitis, allergic hepatitis occurs in a few patients only. Unlike toxic hepatitis, allergic hepatitis is frequently associated with fever, rash or other hypersensitivity manifestations; it may be hepatocellular, mixed or cholestatic; it promptly recurs after inadvertent drug rechallenge. Lysosomal phospholipidosis occurs frequently with three antianginal drugs (diethylaminoethoxyhexestrol, amiodarone and perhexiline). These cationic, amphiphilic drugs may form phospholipid-drug complexes within lysosomes. Such complexes resist phospholipases and accumulate within enlarged lysosomes, forming myeloid figures. This phospholipidosis has little clinical importance. In a few patients, however, it is associated with alcoholic-like liver lesions leading to overt liver disease and, at times, cirrhosis. Subjects with a deficiency in a particular isoenzyme of cytochrome P 450 poorly metabolize perhexiline and are at higher risk of developing liver lesions. Prolonged, drug-induced liver-cell necrosis may also lead to subacute hepatitis, chronic hepatitis or even cirrhosis. This usually occurs when the drug administration is continued, either because the liver disease remains undetected or because its drug aetiology is overlooked. Several autoantibodies may be present.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3044469 TI - Drug-induced cholestasis. AB - Acute, drug-induced hepatocellular cholestasis (either pure or cholestatic hepatitis) is a common manifestation of drug-induced hepatic injury. The drugs most frequently responsible are hormonal steroids and psychopharmacological agents (in particular phenothiazines and some antidepressants). Cholestasis usually subsides without sequelae in less than six months. Acute, drug-induced ductular cholestasis is uncommon and can resemble biliary tract obstruction. Complete recovery occurs promptly after the withdrawal of the causative drug in most cases. The pathogenetic mechanism may be immunoallergic. Prolonged ductular or ductal cholestasis can follow drug-induced acute hepatitis despite prompt withdrawal of the offending drug. This syndrome, observed mainly with chlorpromazine and uncommonly with twenty other drugs, is characterized by the progressive disappearance of small bile ducts and by manifestations mimicking primary biliary cirrhosis. However, its prognosis appears to be better than that of primary biliary cirrhosis, the condition being reversible in the majority of cases or even subsiding completely. The mechanism is still unknown, but several features suggest some form of autoimmunity. Extrahepatic cholestasis related to sclerosing cholangitis is a frequent and long-term complication of intra-arterial infusion of floxuridine in patients treated for hepatic metastases from colorectal carcinoma. Although it may be reversible, floxuridine-induced sclerosing cholangitis has a poor prognosis and can lead to death in a few patients. The mechanism is probably related to the vascular supply of the common hepatic duct and its relationship to the perfusion territory of floxuridine. PMID- 3044471 TI - Drug-induced and toxic granulomatous hepatitis. AB - The ability to induce granulomatous hepatitis has been attributed to numerous drugs; some sixty causative drugs have been culled from the literature for this review. Additionally, granulomas or granulomatoid lesions have resulted from occupational exposure to toxic substances (e.g. silica, copper sulphate, beryllium compounds), and particulate material from various therapeutic or diagnostic procedures (e.g. reactions to starch, talc, suture material, polyvinyl pyrrolidone, silicone, barium sulphate, thorium dioxide) or from intravenous drug abuse (e.g. talc). Clinically, patients with drug-induced or toxic granulomatous hepatitis may be asymptomatic. More frequently, the presentation is that of an acute febrile illness, with or without a rash and eosinophilia, followed by jaundice and biochemical evidence of hepatic dysfunction. The diagnosis of drug induced granulomatous hepatitis is based largely on ruling out other aetiologies. Liver biopsy plays a key role in diagnosis. Recovery is the rule following withdrawal of the drug. Morphologically, drug-induced granulomas may be impossible to distinguish from those due to other causes. Associated lesions suggesting a drug aetiology include significant tissue eosinophilia, unicellular hepatocytic degeneration and necrosis, cholestasis and acute cholangitis or vasculitis. Special stains, polarizing and phase contrast microscopy, transmission and scanning electron microscopy and energy dispersive X-ray microanalysis all play a role in the aetiologic diagnosis of some types of granulomas. PMID- 3044470 TI - Fatty change. PMID- 3044472 TI - Drug-induced vascular and sinusoidal lesions of the liver. AB - Drugs can induce various vascular and sinusoidal lesions of the liver, in particular hepatic vein thrombosis, veno-occlusive disease, sinusoidal dilatation, peliosis hepatis, hepatoportal sclerosis and angiosarcoma. None of these lesions is specific of drug-induced injury and all of them can be determined by causes other than drugs. However, for one of these lesions, veno occlusive disease, chemotherapy (and/or irradiation) represents the main aetiology. PMID- 3044473 TI - The effect of indomethacin and aspirin on alkaline phosphatase secretion and [3H]thymidine incorporation by human osteoblasts. AB - Since it has been suggested that long-term treatment with indomethacin and other non-steroidal anti-inflammatory drugs (NSAIDs) may result in destructive changes in the hip joint, we have examined the effect of indomethacin and aspirin on the secretory and mitotic activity of human osteoblasts in culture. Both indomethacin and aspirin inhibited the secretion of alkaline phosphatase and the uptake of [3H]thymidine by osteoblasts in a dose-dependent fashion at therapeutic concentrations. Both drugs also inhibited insulin- and 1,25 dihydroxycholecalciferol-stimulated alkaline phosphatase production and [3H]thymidine uptake by human osteoblasts. It is concluded that indomethacin and aspirin, and possibly other NSAIDs, may have an inhibitory effect on osteoblast function. PMID- 3044475 TI - Duplex Doppler ultrasound in the diagnosis of cavernous transformation of the portal vein. AB - Duplex Doppler ultrasound scans were performed in 10 patients with proven cavernous transformation of the portal vein. All cases exhibited absence of the normal portal vein lumen with replacement by numerous tortuous vessels. Doppler studies revealed a characteristic "flat" waveform with a reduction in the time averaged velocity to less than 8 cm/s which is well below the normal range. The combination of real-time and Doppler ultrasound enabled an accurate diagnosis to be made in all cases. PMID- 3044474 TI - Treatment with acyclovir does not affect orogenital ulcers in Behcet's syndrome: a randomized double-blind trial. AB - Acyclovir is a potent antiviral drug, of proven efficacy in the treatment of herpes simplex virus infection. Since this virus has been implicated in the aetiology of Behcet's syndrome, the effect of acyclovir on orogenital ulceration in this condition was determined in a randomized, double-blind, placebo controlled, crossover trial. Eighteen of 22 patients entering the study completed the trial. Treatment with acyclovir failed to alleviate the frequency and severity of orogenital ulceration or other disease features. PMID- 3044476 TI - The 10-year results of a prospective trial of post-operative radiotherapy delivered in 3 fractions per week versus 2 fractions per week in breast carcinoma. AB - The 10-year results are presented of a prospective trial of 411 patients with breast carcinoma treated by mastectomy and post-operative radiotherapy given in either 2 or 3 fractions per week (i.e. a comparison of 6 fractions in 18 days with 12 fractions in 28 days). The early radiation effects on the normal tissues were similar and acceptable. The late skin changes in the chest wall (treated with 70 kV X rays) were progressive and by 10 years were slightly more marked with 6 fractions. Late subcutaneous fibrosis in the axilla (treated with cobalt 60 teletherapy), however, was much less in the 6-fraction group. Twelve fractions resulted in greater restriction of shoulder movement and an increased incidence of lymphoedema of the arm. Doses were selected on the basis of past clinical experience. The dose used to treat the axilla in 6 fractions was 35 Gy, 14.99% less than that predicted by NSD. The dose predicted by alpha:beta, using a value of 2 Gy for late reactions, is 38.14 Gy. Thus simple theory, which omits time, still predicts too high a dose for 6 fractions, although it is closer than NSD. In this trial, the 6-fraction technique showed an advantage over the 12-fraction technique. It was equally effective in controlling local recurrence and had fewer late sequelae. It was also convenient for patients and economic in the use of radiotherapy resources. PMID- 3044477 TI - The cost of imaging procedures. PMID- 3044478 TI - Neurophysiological measurements in patients with genuine stress incontinence of urine and the relation of neurogenic defects to the presence of spina bifida occulta. AB - Neurophysiological measurements comprising surface electromyography (EMG) of the urethral and anal sphincters, measurement of the sensory thresholds of the dorsal nerve (DN) of the clitoris and the urethra and sacral reflex latencies from the dorsal nerve to urethra and anus and from the urethra to anus were performed in 44 females with genuine stress incontinence, in order to detect any relationship between the clinical disorder and a neurogenic defect in the innervation of the muscles involved in maintaining continence. Plain X-rays of the renal tracts (KUB films) were also studied to establish whether the prevalence of spina bifida occulta was increased in this group of patients and was a likely explanation for the nerve defects. Forty-three patients were found to have abnormal neurophysiological responses and the prevalence of spina bifida occulta was 50%, which compares with a prevalence of 17% in normal female controls. Genuine stress incontinence appears to be at least partly associated with defective innervation of the lower urinary tract and it is likely that in some cases the defect may be related to the presence of spina bifida occulta. PMID- 3044479 TI - Ureteric obstruction secondary to pelvic actinomycosis. PMID- 3044480 TI - Multilocular hydrocele after sclerotherapy. PMID- 3044481 TI - Prognostic indices in transitional cell carcinoma of the bladder. PMID- 3044482 TI - The value of sonography in the diagnosis and follow-up of patients with blunt renal trauma. AB - Eighty-eight patients with blunt renal trauma were examined sonographically. Rupture of the kidney was diagnosed in 41 patients and contusion of the kidney in 45. The correlation between operative findings, sonography and urography showed positive sonographic findings to be correct in all patients with rupture of the kidney. In 4 patients with normal sonograms, the diagnosis of a contusion was based on the urographic results. Three patients underwent angiography. An intimal lesion was seen in 2 and a tear of the renal artery in 1. The results of our investigation suggest that sonography should be applied in the first place in the evaluation of blunt renal trauma. It is the investigation of choice during follow up after surgical as well as conservative therapy. PMID- 3044484 TI - Cephradine prophylaxis in transurethral procedures for carcinoma of the bladder. AB - The value of cephradine prophylaxis in reducing urinary infection was assessed in 243 patients undergoing endoscopy for bladder carcinoma. Patients were randomised either to receive 3 peri-operative doses of cephradine or to receive no antibiotic. Urine specimens taken on the fifth post-operative day showed a significantly lower urinary infection rate in those patients receiving cephradine prophylaxis. PMID- 3044485 TI - Traumatic rupture of the testis. AB - We present 6 cases of traumatic rupture of the testis. Early scrotal exploration is indicated in this condition and may involve either repair or removal of the injured testis. The extent of microvascular damage to the testis determines the outcome of a repair operation. PMID- 3044483 TI - Ureteric obstruction in stented renal transplants. AB - The use of renal stents in renal transplants is becoming increasingly popular and has been routine practice at this institution for the past 4 years. Despite this procedure, however, obstruction of the transplant ureter still occurs and we describe 7 patients who developed renal obstruction in the presence of a stented transplant ureter. The underlying causes of these findings are discussed together with suggested recommendations for optimum radiological assessment of transplant ureteric stents. The implication of these observations is that urine may flow down stented ureters between the ureter and the stent rather than down the stent lumen. PMID- 3044486 TI - Relation between falciparum malaria and HIV seropositivity in Ndola, Zambia. PMID- 3044488 TI - The fall and rise of general practice. PMID- 3044487 TI - Forty years of the NHS. Origins and early development. PMID- 3044490 TI - Free health care--at a price. PMID- 3044489 TI - Doctor-patient relationships--as close as ever. PMID- 3044491 TI - Forty years of the NHS. Interviews with consultants. PMID- 3044492 TI - Personal views of the NHS--warts and all. PMID- 3044493 TI - Forty years of the NHS. The act, the minister, and the editors. PMID- 3044494 TI - ABC of eyes. The eye and the nervous system. PMID- 3044495 TI - Obstructive uropathy. PMID- 3044496 TI - Adhesive Escherichia coli in inflammatory bowel disease and infective diarrhoea. AB - The clinical features of ulcerative colitis and Crohn's disease are similar to those of infections of the bowel, although their cause is uncertain. Many bacteria that cause intestinal diseases adhere to the gut mucosa, and adhesion of pathogenic Escherichia coli is resistant to D-mannose. The adhesive properties of isolates of E coli were assessed by assay of adhesion to buccal epithelial cells with mannose added. The isolates were obtained from patients with inflammatory bowel diseases (50 with a relapse of ulcerative colitis, nine with ulcerative colitis in remission, 13 with Crohn's disease, and 11 with infectious diarrhoea not due to E coli) and 22 controls. The median index of adhesion to buccal epithelial cells (the proportion of cells with more than 50 adherent bacteria) for E coli from patients with ulcerative colitis in relapse was significantly higher (43%) than that for controls (5%) and patients with infectious diarrhoea (14%). The index was not significantly different among isolates from patients with ulcerative colitis in relapse, Crohn's disease (53%), and ulcerative colitis in remission (30%). If an index of adhesion of greater than 25% is taken as indicating an adhesive strain 86% of isolates of E coli from patients with inflammatory bowel disease were adhesive compared with 27% from patients with infective diarrhoea and none from controls. The adhesive properties of the isolates from patients with inflammatory bowel disease were similar to those of pathogenic intestinal E coli, raising the possibility that they may have a role in the pathogenesis of the condition; the smaller proportion of adhesive isolates in patients with infective diarrhoea due to other bacteria suggests that the organism may be of primary importance rather than arising secondarily. PMID- 3044497 TI - ABC of eyes. Injuries to the eye. PMID- 3044498 TI - Consensus conference. Treatment of stroke. PMID- 3044499 TI - Bovine spongiform encephalopathy. PMID- 3044500 TI - Modern treatment of heart failure. PMID- 3044501 TI - Submucous cleft palate. PMID- 3044502 TI - Prospects for vaccines against HIV. PMID- 3044504 TI - Treating asthma in preschool children. PMID- 3044505 TI - Structured abstracts. PMID- 3044503 TI - The case for low dose diuretics in hypertension: comparison of low and conventional doses of cyclopenthiazide. AB - In a double blind placebo controlled randomised parallel study the antihypertensive activity and adverse biochemical effects of three doses of cyclopenthiazide were evaluated in patients with mild essential hypertension that had been recently diagnosed or was being treated with a single drug. After a four week placebo washout period 53 patients with diastolic blood pressures between 90 110 mm Hg were randomly assigned to 50, 125, or 500 micrograms cyclopenthiazide or matching placebo for an eight week period of treatment. Blood pressure was measured in the patients' homes by the same observer every two weeks. Serum urea, electrolytes, urate, and creatinine concentrations and 24 hour urinary sodium excretion were monitored every four weeks and serum magnesium concentration and plasma renin activity at the end of the washout and treatment periods. After eight weeks of treatment systolic and diastolic blood pressures were significantly reduced in patients taking 125 and 500 micrograms cyclopenthiazide when compared with those taking placebo. The decrement in serum potassium concentration (0.6 mmol/l) and increase in serum urate concentration 0.06 mmol/l) were greatest with the 500 micrograms dose, the increase in serum urate concentration alone being significant. No change in serum magnesium concentration or 24 hour urinary sodium excretion was noted with any dose of cyclopenthiazide. Only the 500 micrograms dose of cyclopenthiazide significantly increased the mean plasma renin activity (1.8 (95% confidence interval 0.2 to 3.4)-5.4 (3.9 to 6.8) nmol angiotensin I/l/h); the other doses like the placebo had no effect. Cyclopenthiazide 125 micrograms, a dose lower than is currently marketed, produced a similar hypotensive response to 500 micrograms of the drug without upsetting the biochemical profile. PMID- 3044506 TI - Controlled trial of budesonide given by the nebuhaler in preschool children with asthma. AB - OBJECTIVE: To determine whether the inhaled corticosteroid budesonide, given by a Nebuhaler spacing device, was effective in prophylaxis of asthma in preschool children. DESIGN: Double blind, placebo controlled, random order crossover trial with two week practice run in period. SETTING: Outpatient clinic referrals in secondary referral centre. PATIENTS: 39 children aged 2-6 years selected for the following: able to use Nebuhaler; parents able to complete record card; poorly controlled asthma (defined); not already on systemic or inhaled steroids. Eleven withdrew for various reasons not connected with intolerance to budesonide. Age, sex, other atopies, and symptoms during run in period were similar in the 28 children who completed the trial and in the 11 who withdrew. INTERVENTIONS: Budesonide 200 micrograms or placebo (both one puff) given twice daily during 6 week treatment or control periods, using Nebuhaler after prior training. Three week "washout" at crossover. Compliance monitored by weighing canisters. Patients withdrawn if their acute attacks required treatment with systemic steroids. END POINT: Control of asthma. MEASUREMENTS AND MAIN RESULTS: Peak expiratory flow rate measured twice daily where cooperation allowed. Diary of symptoms and concomitant drug use kept daily. Results showed mean peak flow significantly higher (12% in mornings, 14% in evenings) in second three weeks of intervention compared with control period (95% confidence intervals 6.3-17.3% and 7.2-21.0%). Supplementary bronchodilator drugs reduced by 50% during intervention periods. CONCLUSIONS: Budesonide given by Nebuhaler is effective prophylaxis for preschool children with frequent asthma. PMID- 3044507 TI - Flosequinan in heart failure: acute haemodynamic and longer term symptomatic effects. AB - There is no single, simple test with which to evaluate new treatments for heart failure. Various methods need to be used, and a study of both the acute haemodynamic and longer term symptomatic effects of flosequinan, a new direct acting arteriolar and venous vasodilator, was therefore carried out in patients with heart failure. In one group of patients flosequinan increased cardiac output and caused a fall in pulmonary capillary wedge pressure, both effects lasting for 24 hours. In a double blind, placebo controlled study in another group flosequinan improved mean exercise tolerance from 9.9 to 12.7 minutes after four weeks of treatment. The drug also reduced perceived exertion during submaximal exercise and increased calf and therefore skeletal muscle blood flow. It reduced plasma renin activity and noradrenaline concentrations. Flosequinan possesses all the important properties of a drug likely to be of value in the treatment of heart failure. PMID- 3044508 TI - Combination treatment with cholestyramine and bezafibrate for heterozygous familial hypercholesterolaemia. AB - Cholestyramine and bezafibrate were compared individually and in combination in the treatment of 18 patients with heterozygous familial hypercholesterolaemia. The study used a double blind, placebo controlled block design with a placebo run in period of two months followed by three phases of active treatment, each of two months' duration. Patients were randomly allocated to one of the six possible sequences of medication so that three patients would be treated with each sequence. Two patients withdrew from the study before completion. The median concentration of total cholesterol decreased from 9.65 mmol/l (interquartile range 8.62 to 8.72) to 7.24 mmol/l (6.70 to 7.52) with cholestyramine, to 8.09 mmol/l (7.18 to 8.68) with bezafibrate, and to 6.31 mmol/l (5.84 to 7.27) with the combination. This fall was due almost entirely to a decrease in the low density lipoprotein cholesterol concentration, and the combination was significantly more effective than either drug alone. The 98% confidence intervals for the median differences between the combination and cholestyramine and the combination and bezafibrate were 0.04 to 1.49 mmol/l and 0.51 to 2.18 mmol/l respectively. These results suggest that this combination is an effective and useful treatment in heterozygous familial hypercholesterolaemia. PMID- 3044510 TI - ABC of eyes. Refractive errors. PMID- 3044511 TI - Harvesting organs for transplantation. PMID- 3044509 TI - Fish oil and plasma fibrinogen. PMID- 3044512 TI - Towards bloodless liver surgery. PMID- 3044513 TI - Prophylactic sclerotherapy for varices. PMID- 3044514 TI - Pneumocystis carinii pneumonia: detection of parasites in sputum and bronchoalveolar lavage fluid by monoclonal antibodies. AB - Diagnosis of pneumocystis pneumonia is based on identifying Pneumocystis carinii cytochemically in material from the lung. The silver methenamine staining methods most commonly used are technically difficult and lack specificity. The diagnostic value of immunocytological identification of the parasite was evaluated by using mouse monoclonal antibody 3F6, specific for human pneumocystis, to identify P carinii in bronchoalveolar lavage fluid and sputum by immunofluorescence and was compared with that of other variables. Bronchoalveolar lavage was performed on 25 patients positive for HIV antibody with clinically suspected pneumocystis pneumonia and 40 patients negative for HIV antibody who presented with interstitial disorders of the lung. Lavage fluid showed pneumocystis only in the patients positive for antibody, the parasite being detected in 19 by immunofluorescence and in 17 by a modified silver methenamine staining method. Chest x ray films obtained at the time of bronchoscopy showed interstitial or alveolar shadowing in 17 of the 19 patients, but clinical symptoms and the presence of antibodies to pneumocystis did not seem to be predictive. Sputum samples were collected during 43 episodes of clinically suspected pneumocystis pneumonia in patients positive for HIV antibody. Pneumocystis was detected consistently more commonly by immunofluorescence than the silver strain in sputum collected routinely and induced by inhalation of saline. In 17 patients bronchoalveolar lavage followed sputum collection, and the sensitivity of detection of pneumocystis in immunofluorescence in sputum compared with lavage fluid was 57% (8/14). Immunofluorescence was suitable for specimens fixed in ethanol and seemed highly specific and more sensitive than the standard cytochemical methods for identifying pneumocystis. PMID- 3044516 TI - The release of pituitary LH and sprouting of LH-releasing hormone (LHRH) containing neurons after anterior hypothalamic deafferentation. AB - The chronology of changes in plasma luteinizing hormone (LH) and the distribution of immunoreactive neuronal processes containing LH releasing hormone (LHRH-ir) were studied in the female rat after surgical interruption of anterior neural connections of the mediobasal hypothalamus (MBH). Spontaneous LH surges on the afternoon of proestrus and LH release after estradiol benzoate (EB) followed 48 h later by progesterone (P) administration were studied in ovariectomized (OVX) rats. The maximum increase in plasma concentrations (delta maxLH) after EBP was calculated for each rat at several intervals over 140 days. Control animals given EBP at monthly intervals after OVX had comparably large delta maxLH surges during the first few months of study. However, a gradual decline in control delta maxLH followed becoming significant 3 months after the start of the experiment. In contrast, frontal cuts (FC), which interrupted anterior MBH connections, produced an abrupt decrease in delta maxLH surges after EBP to 11% of preoperative levels. However, during subsequent EBP trials, there was a gradual improvement in LH surges to about 50% of preoperative levels over 100 days. In some cases, individual improvement became equal to preoperative LH surge levels, in others there was no recovery. Examination of LHRH-ir nerve fiber growth responses after FC suggested that sprouting by these peptide-containing neuronal processes may have contributed to the functional improvements observed. PMID- 3044515 TI - Comparison of mosquito nets, proguanil hydrochloride, and placebo to prevent malaria. AB - One hundred and ninety students aged 6 to 18 at a boarding school 120 km west of Nairobi in the Rift Valley participated in a comparative trial of malaria prophylaxis. Treatment with a combination of amodiaquine 25 mg/kg over three days plus doxycycline 100 mg twice daily for five days cleared their blood of Plasmodium falciparum. They were then randomly divided into the following three groups matched for age and sex: one group slept under mosquito nets; one group received one or two tablets (100 mg each) of proguanil hydrochloride daily according to weight; one group received one or two placebo tablets daily which were the same size and colour as the proguanil tablets. Malaria was diagnosed when asexual P falciparum were seen on blood films and was treated with pyrimethamine-sulphadoxine. At the end of one school term 188 of the 190 students had completed the study. One new infection was found during 3893 days of follow up in the mosquito net group, eight new infections over 3667 days in the proguanil group, and 35 new infections over 3677 days in the placebo group, representing a reduction of 97.3% and 77.1% in attack rates for the mosquito net method and for treatment with proguanil respectively. Both provide effective protection from malaria. PMID- 3044517 TI - The history of thought concerning the hypothalamus and its functions. AB - Many initial studies related to identification of the boundaries and structural components, nuclei, tracts and interconnections of the hypothalamus; this continues. Early interest also focused on hypothalamic control of somatic activities and autonomic nervous system functions. During the present century chiefly, interest has developed in the hypothalamus and control of water balance, thirst, water retention and loss (diabetes insipidus and polydipsia). Its role in control of metabolism, body weight (obesity), and the regulation of body temperature has attracted the attention of physiologists for many years. Others have studied hypothalamic regulation of sex and reproductive phenomena. The hypothalamus is now attracting much attention because of its production of neuroendocrine secretions and role in control of the endocrine system. Physiologists realized very early that the hypothalamus is involved in emotional expression, in reaction to stress and adaptive adjustments. Its involvement in disease states and resistance thereto and in determining the nature of behavior has now been recognized as a matter of great importance. The origins of all these interests are reviewed. PMID- 3044518 TI - Synaptic and neuronal-glial plasticity in the adult oxytocinergic system in response to physiological stimuli. AB - Magnocellular oxytocinergic neurons in the hypothalamus offer a striking example of a mammalian neuronal system whose basic architecture and synaptic circuitry can be reversibly modified in adulthood. During parturition, lactation and prolonged osmotic stimulation, glial coverage of oxytocinergic neurons markedly diminishes and their surfaces are left in extensive juxtaposition; concurrently, there is formation of new synapses, which are predominantly GABAergic and which couple two or more oxytocinergic neurons simultaneously. These structural changes do not permanently modify the anatomy of the system since upon cessation of stimulation, neuronal juxtapositions and shared synapses disappear, to reappear upon new stimulation. At present, we can only speculate about the cellular mechanisms and factors responsible for these reversible neuroanatomical changes. However, oxytocin itself appears to be of primary importance since it can induce similar anatomical changes when chronically infused into the third ventricle. PMID- 3044519 TI - Dynamism of chemoarchitecture in the hypothalamic paraventricular nucleus. AB - The hypothalamic paraventricular nucleus (PVN) has been implicated in a remarkable number of functions including control of pituitary-adrenocortical activity in response to stress, body fluid homeostasis, milk ejection reflex, prolactin secretion, thyroid hormone secretion, analgesia, food intake, gastrointestinal functions, cardiovascular functions, and control of pineal melatonin synthesis. Paraventricular neurons produce hormones of key importance in neuroendocrine regulation such as vasopressin (VP), oxytocin (OX), 41-residue corticotropin releasing factor (CRF), thyrotropin releasing hormone (TRH), somatostatin (SOM) and the putative prolactin releasing factor vasoactive intestinal polypeptide (VIP). Three recent advances pertinent to the organization of the PVN include: (1) the evidence that the structure of the PVN is compartmental in nature, topographically segregated cellular units seem to carry out different functions; (2) the discovery that paraventricular neurons are capable of expressing a multitude of neuromediators simultaneously, thus cellular units can be best specified by a certain combination of neuromediators; (3) evidence that the composition of the neuromediator "cocktail" in individual neurons is variable and depends on the physiological status of the animal. Hence, the PVN may be best considered as a dynamic mosaic of chemically specified subgroups of neurons. The flexibility of neurotransmitter status in paraventricular neurons may play a central role of a functional plasticity of fixed anatomical circuits. PMID- 3044520 TI - The Pro-LHRH system of the rat brain. Effects of changes in the endocrine background. AB - The gonadotropin-releasing hormone-associated peptide (GAP) and luteinizing hormone-releasing hormone (LHRH) portions of the LHRH precursor were localized by immunocytochemistry in prepubertal female rats, in adult female rats at different stages of the estrous cycle, and in ovariectomized rats. Our results indicate that GAP is present in the same population of neurons as LHRH in the rat brain. These results confirm the specificity of previous immunocytochemical studies which used antisera to LHRH alone. The endocrine status of the animal was demonstrated to affect the immunocytochemical appearance of the GAP system. The number of GAP immunopositive cells and terminals is highest during diestrus II and lowest on the day of estrus, suggesting either a role in and/or a dependence upon the endocrine changes associated with the estrous cycle. Ovariectomy results in a gradual decrease in GAP immunoreactivity in the median eminence. This observation, in concert with other recent studies, suggests that ovarian factors may be acting to maintain the LHRH system and that ovariectomy may result in decreased synthesis and/or processing of the LHRH system and that ovariectomy may result in decreased synthesis and/or processing of the LHRH precursor. PMID- 3044521 TI - Role of central oxytocin in the control of the milk ejection reflex. AB - The neuropeptide oxytocin, synthetized by magnocellular neurons in the hypothalamus, is well known for its peripheral action after it is released into the bloodstream from axons in the neurohypophysis. Less familiar is the notion that it is also released centrally to control the activity of oxytocinergic neurons themselves. When injected into the third ventricle of lactating rats during suckling, oxytocin increases the basal firing rate of oxytocinergic neurons as well as their activity at the time of each reflex milk ejection. On the other hand, centrally administered oxytocin engenders the neuronal-glial and synaptic plasticity characteristic of the oxytocin system when it is physiologically activated. From numerous in vivo and in vitro observations, it appears that central oxytocin is released in the hypothalamic nuclei themselves. For example, the use of push-pull cannulae inserted into one supraoptic nucleus of suckled rats shows that oxytocin is released inside the nucleus specifically during milk ejection. Moreover, ultrastructural immunocytochemistry reveals synaptic terminals in the supraoptic nucleus where both the pre- and postsynaptic elements are oxytocinergic. Nevertheless, the mechanism of the central release of the neuropeptide has still to be determined, especially in view of electrophysiological observations indicating that the release process in the hypothalamus is different from that within the neurohypophysis. PMID- 3044523 TI - Cardiovascular input to hypothalamic neurosecretory neurons. AB - In vivo extracellular recordings from rat supraoptic and paraventricular magnocellular neurosecretory cells (MNCs) indicate that putative vasopressin secreting MNCs may be identified by an abrupt and brief cessation in firing consequent to a transient drug-induced rise in arterial pressure sufficient to activate arterial baroreceptors. In the diagonal band of Broca (DBB), a population of neurons projecting towards the supraoptic nucleus are activated during this drug-induced hypertension. Electrical stimulation in DBB selectively depresses supraoptic vasopressin-secreting MNCs. Intracellular recordings in perfused hypothalamic explants confirm a DBB-evoked bicuculline-sensitive and chloride-dependent postsynaptic inhibition, similar to that associated with the application of gamma-aminobutyric acid (GABA) in approximately half of supraoptic MNCs. Since bicuculline also selectively blocks baroreceptor-induced inhibition in supraoptic MNCs, it is proposed that the depressant baroreflex input to vasopressin-secreting MNCs involves a population of DBB neurons and GABAergic interneurons located close to MNCs. An excitatory and selective input to vasopressin-secreting MNCs follows chemoreceptor activation, possibly mediated by the A1 noradrenergic cell group in the ventrolateral medulla. Another excitatory input to both vasopressin- and oxytocin-secreting MNCs is triggered by circulating angiotensin II and appears to be relayed centrally through an angiotensinergic projection from the subfornical organ. PMID- 3044522 TI - Synaptic inputs and electrical coupling among magnocellular neuroendocrine cells. AB - This paper first briefly reviews the evidence for synaptic and nonsynaptic plasticity among the neurons and glia of the magnocellular hypothalamo neurohypophysial system. Emphasis is placed upon the importance of the roles played by astrocytes in the remodeling of the magnocellular nuclei under various conditions of increased hormone demand. Evidence is then reviewed from more recent studies showing that there is electrical coupling among magnocellular neurons, and that this coupling shows plasticity similar to that shown for other characteristics of the system (e.g., chemical synapses, dendritic bundling etc.). Further, evidence is presented that extent of electrical coupling can be modified not only by manipulating the physiological state of the animal (such as lactation), but also by electrical stimulation of newly described olfactory afferent inputs to the cells of the supraoptic nucleus. The possible functional significance of these findings is discussed in relation to the behavior of nursing rats. PMID- 3044524 TI - Hypothalamic control of gastric acid secretion. AB - Study of hypothalamic control of gastric acid secretion (GAS) has revealed GAS related neurons, their location in the lateral hypothalamic area (LHA), their characteristics, and implications of their relations to feeding and other functions. Some LHA glucose-sensitive neurons are referred to as gastric type because of their effects on gastric oxyntic cells via specific gastric related neurons of the medulla oblongata and the vagus. The 2-deoxy-D-glucose (2-DG), or insulin induced GAS was completely abolished by bilateral subdiaphragmatic vagotomy, or micro-lesions in specific sites of the LHA. These gastric type glucose-sensitive neurons were thus believed to contribute to control of GAS. The paraventricular nucleus (PVN) was also found to affect GAS. GAS-related PVN neurons were observed in the rostral PVN. Electrophoretic application of various chemicals, especially glucose, also affected neurons in the rostral PVN. Electrophoretically applied norepinephrine (NE) increased PVN single neuron activity and suppressed GAS. Results suggest that the rostral PVN may be another site to modulate LHA control of GAS, and NE may be a transmitter or modulator. PMID- 3044525 TI - The role of the AV3V region in the control of magnocellular oxytocin neurons. AB - The AV3V region is important in the control of body fluid and Na+ regulation and projects to the supraoptic and paraventricular nuclei. Oxytocin from the neurohypophysis mediates milk ejection and is involved in parturition, but has also been recently implicated as a candidate natriuretic hormone. We have studied the role of the AV3V region in the control of magnocellular oxytocin neurons in rats. Electrical stimulation of the AV3V region increased the firing rate of supraoptic oxytocin neurons and evoked a concomitant release of oxytocin. Acute electrolytic AV3V lesions silenced supraoptic neurons and abolished their excitation by hyperosmotic stimulation. The lesions also abolished osmotically induced release of oxytocin. Re-activation of supraoptic neurons by local glutamate restored their osmoresponsiveness to about 50% normal. Thus, while supraoptic neurons are directly osmosensitive, the AV3V region is essential for their normal osmoresponsiveness. Electrolytic AV3V lesions did not affect suckling-induced oxytocin secretion or, in conscious rats, the release of oxytocin secretion during parturition. Thus the AV3V region is not involved in the activation of oxytocin neurons during suckling or parturition. PMID- 3044526 TI - Neuronal sensitivities in preoptic tissue slices: interactions among homeostatic systems. AB - The preoptic area participates in many homeostatic systems, which include the regulation of body temperature, fluid and metabolite balance, and reproduction. Some preoptic neurons have been shown to be sensitive to either temperature, osmotic pressure, glucose, testosterone or estradiol. While previous studies have treated these as separate and distinct neuronal populations, this paper reviews recent experiments which show that many neurons have multiple sensitivities to these endogenous factors. Neurons in preoptic tissue slices were tested for their responses to changes in temperature, as well as various perfusion media containing 30 pg/ml testosterone or estradiol, low glucose (1.0 mM) or increased osmotic pressure (309 mosmol/kg). The steroid-sensitive, osmosensitive and glucosensitive neurons were not confined to the temperature insensitive neurons; but instead nearly half of the thermosensitive neurons responded to these nonthermal stimuli. In addition, many osmosensitive neurons showed glucosensitivity and steroid-sensitivity. This suggests that, even at the neuronal level, there is a basis for interactions between homeostatic systems. PMID- 3044527 TI - Convergence of thermal, osmotic and cardiovascular signals on preoptic and anterior hypothalamic neurons in the rat. AB - Responsiveness of thermosensitive neurons in the preoptic and anterior hypothalamus (PO/AH) to osmotic and cardiovascular signals have been shown to be responsible, at least partly, for the reduced thermoregulation during dehydration and the hypothermia after acute blood loss. The responsiveness to local and peripheral (hepatoportal) osmotic stimuli were found in about 60% of PO/AH thermosensitive neurons and 12% of thermally insensitive neurons in tissue slices in vitro and in urethane-anesthetized rats. Since hyperosmotic stimuli predominantly decreased the activity of both warm-sensitive and cold-sensitive neurons, the reduced heat loss and heat production during dehydration may be explained by altered activity of PO/AH thermosensitive neurons induced by hyperosmolality. About 42% of 250 PO/AH neurons (66.3% of thermosensitive neurons and 30% of thermally insensitive neurons) exhibited the responsiveness to changes in blood pressure by less than 15 mmHg, which was found to be mediated by baro/volume receptors. Hypotensive stimuli predominantly increased the activity of warm-sensitive neurons and decreased the activity of cold-sensitive neurons. The neuronal responses may explain, at least in part, the hypothermia after acute bleeding. PMID- 3044528 TI - The role of vasopressin as an antipyretic in the ventral septal area and its possible involvement in convulsive disorders. AB - Perfusion of the peptide, arginine vasopressin (AVP), within the ventral septal area (VSA) of the brain of a number of species reduces fever but not normal body temperature. This antipyretic response appears to be mediated by AVP receptors of the V1 subtype. Lesions of the VSA with kainic acid are associated with prolonged and enhanced fevers in rats. A role for endogenous AVP in fever suppression within the VSA comes from several types of experiments: (1) AVP release within the VSA is inversely correlated to fever height; (2) AVP antagonists or antiserum injected into the VSA prolong fever; (3) animals lacking endogenous AVP in the VSA (Brattleboro rat, long-term castrated rat) develop enhanced fevers. Electrical stimulation of the AVP-containing cell bodies of the bed nucleus of the stria terminalis (BST) orthodromically inhibits VSA neurons and also suppresses fever; the latter effect can be abolished with application of a V1 antagonist to the VSA. Iontophoretic studies indicate that AVP inhibits glutamate stimulated activity of thermoresponsive and other VSA neurons. AVP can also act in the VSA to cause severe motor disturbances; this action is receptor mediated and increases in severity upon sequential exposure to AVP. Because sites of action of the antipyretic and convulsive action of AVP are similar, and because animals lacking brain AVP display reduced convulsive activity, it is possible that AVP, released during fever, could be involved in the genesis of convulsive activity. PMID- 3044529 TI - [Processing and presentation of antigens as a part of the induction of the immune response]. PMID- 3044530 TI - [The role of serum factors in phagocytosis of Candida albicans by rabbit macrophages]. PMID- 3044531 TI - Effect of Escherichia coli endotoxin on tissue lipoprotein lipase activities in chickens. AB - 1. Four-week-old broiler chickens were injected intravenously with from 0.01 to 1 mg of E. coli endotoxin/kg body weight or with saline. 2. At all doses used endotoxin markedly depressed food intake and lipoprotein lipase activities in muscle and adipose tissue within 8 h. Heart lipoprotein lipase activity was significantly depressed only at doses of 0.1 mg endotoxin/kg body weight or greater. 3. Treatment of birds with 0.3 mg endotoxin/kg body weight reduced post heparin lipoprotein lipase activity to 0.13 of that in control birds in 8 h. 4. Endotoxin generally depressed plasma very-low-density lipoprotein concentration. Plasma non-esterified fatty acid concentration was significantly elevated only in birds given 1 mg endotoxin/kg body weight. 5. Fatty acid synthetase activity in the liver of endotoxin-treated birds was significantly lower than in control birds 16 h after administration of endoxin, but not after 8 h. 6. These results show that tissue lipoprotein lipase activity in birds is very responsive to E. coli endotoxin, as in mammals. Hypertriglyceridaemia occurs only occasionally in endotoxin-treated chickens, most probably because of the particularly close relationship between food intake and hepatic lipoprotein synthesis in birds. PMID- 3044532 TI - Prostaglandins and leukotrienes as mediators of shock and trauma. PMID- 3044533 TI - Reproductive physiology of the nonpregnant mare. An overview and update. AB - This article reviews the reproductive events in the nonpregnant mare with emphasis on recent advances. The discussion is restricted to the salient features of puberty (prenatal and prepubertal events), seasonality (gonadotropins, photoperiod, and other modifying factors), and the estrous cycle (hormones, estrus, diestrus, and the control of cyclicity) in the nonpregnant mare. PMID- 3044534 TI - Control of the estrous cycle in the mare. AB - All current approaches to manipulating the reproductive biology of the nonpregnant mare are discussed. PMID- 3044535 TI - Ultrasonic imaging of equine ovarian follicles and corpora lutea. AB - One of the most profound theriogenology applications of transrectal diagnostic ultrasonography in mares involves the imaging of ovarian follicles and corpora lutea. The resolving capabilities (frequency) and quality of the scanner directly affect the minimal size of a structure that can be imaged and the quality of the image. High-frequency scanners (5 or 7.5 MHz) of good quality can image a 2-mm follicle and the corpus luteum throughout its functional life. A low-frequency scanner (3 or 3.5 MHz) can image a 6-mm follicle and the corpus luteum for several days after ovulation. Equine follicles are excellent subjects for transrectal imaging because they are large, filled with fluid, and readily accessible. Event the small follicles (less than 10 mm) can be diagnostically important in evaluating whether ovarian infertility has occurred and whether the follicles are responding to treatment for follicular stimulation. The large, preovulatory follicles are of special interest. Averaged over a group of 79 periods, the following significant changes were found in the preovulatory follicle: increasing diameter, shape change from spherical to pear-shaped or conical, and increasing thickness of the follicular wall. No significant changes were found in the echogenicity (gray-scale value) of the wall or fluid. In retrospect, the diameter of the follicle seemed as useful for predicting impending ovulation as any of the other ultrasound criteria. The occurrence of ovulation is readily detected by the disappearance of a large follicle that was present at a recent previous examination. In addition, the ovulation site on the day of ovulation is detectable. In one study, the site was correctly identified in 24 of 24 mares. A small amount of residual follicular fluid can sometimes (7 of 10 in one study) be detected at the site of ovulation. The residual fluid usually disappears over a period of 0.5 to 20 hours. Subsequently, the developing corpus luteum may form a central nonechogenic area with peripheral luteinization or may remain uniformly luteinized. The central areas are of apparently vascular origin (blood or a component of blood) and become clotted and organized. In one study, approximately 50 per cent of the glands developed central areas exceeding 10 per cent of the size of the gland. The central areas began to develop on Day 0 or 1 and continued to enlarge until Day 2 or 3. The relative proportion of the gland containing a central clot decreases after Day 3, but the central area usually remains visible throughout diestrus.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3044536 TI - Diagnosis and correction of twin pregnancy in the mare. AB - Reproductive Physiology 1. Twin pregnancies result in high rates of abortion, stillbirth, and neonatal mortality. 2. Twins develop subsequent to multiple ovulations. Multiple ovulations are related to breed, parity, and mare history. Multiple ovulations are most frequently seen in Thoroughbred and Draft mares. Multiple ovulations are more common in barren and perhaps maiden mares than in lactating mares, and they are more common in certain individual mares. 3. Equine embryos are motile in the uterus from the time of first detection (Days 9 to 10) until fixation (Day 16). They are frequently located in the uterine body on Days 9 and 10. 4. Twin embryos have a pattern of motility and fixation similar to that of single embryos, and fixation is more frequently unilateral than bilateral (70 per cent versus 30 per cent, respectively). 5. Mares have an efficient natural embryo-reduction mechanism to eliminate excess (greater than 1) embryos resulting from multiple ovulations. Natural embryo reduction is more successful in unilateral than bilateral twin pregnancies (89 per cent versus 11 per cent successful reduction, respectively). 6. After the establishment of endometrial cups (Days 35 to 40), mares that are aborted will frequently not cycle for several months. Management of Twin Pregnancy 1. Breed all mares regardless of the number of preovulatory follicles. Withholding mares with preovulatory follicles from breeding does not decrease the incidence of twin pregnancy, but it decreases the overall pregnancy rate and results in a loss of breeding time. 2. Check all mares for twins, regardless of the number of detected ovulations.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3044537 TI - Uterine defense mechanisms in the mare. AB - Uterine defense against infection in the mare has been actively investigated over the past decade. Mechanisms of defense, including the role of immunoglobulins, polymorphonuclear neutrophils, and the physical ability of mares to eliminate bacteria from the uterus, are discussed. PMID- 3044538 TI - Endometrial biopsy of the mare. A review and update. AB - The endometrial biopsy is a safe and effective means of predicting a mare's prognosis for foaling. A thorough understanding of the normal cyclic and seasonal pattern displayed by the normal endometrium is necessary before interpreting pathologic changes. Several systems for prognostic classification have been proposed, including a recent one that combines many of the criteria used in the other systems. PMID- 3044539 TI - Equine endometrial cytology. AB - The simplicity of collection of material for cytologic preparations belies the complexity of smear interpretation beyond recognition of neutrophils. Knowledge concerning cancer cytology moves rapidly, for cancer is a progressive, often fatal disease so that tissue for comparison and confirmation of interpretation often becomes available. This is not true for cytologic study of the equine endometrium. Lesions detected by means of cytology smears may be transient and regress, offering little information concerning their etiology or consequences. They may be focal and missed in the corresponding biopsy. Such experiences should lead to the abandonment of equine endometrial cytology, but have only strengthened our interest and enthusiasm for this technique in the study of the uterus. We have found it to be useful in a limited number of clinical circumstances in which other techniques have failed. This, plus the growing number of supportive clinicians using cytopathology service and the large number of mares with fertility problems, leads us to believe that further investigation of equine endometrial cytology may prove to be even more helpful as a clinical tool. PMID- 3044540 TI - Embryonic loss in mares. Incidence, possible causes, and diagnostic considerations. AB - Fertilization rates were similar for normal and subfertile mares, and much of the difference in fertility between normal and subfertile mares was due to embryonic loss. Fertilization rate estimates for mares ranged from 71 to 96 per cent. The incidence of embryonic loss detected by ultrasonography between Days 11 and 50 was approximately 9 per cent for normal mares, and the estimated incidence of embryonic loss before Day 14 was also 9 per cent. Therefore, the estimated incidence of embryonic loss in normal mares between fertilization and Day 50 is approximately 18 per cent (Fig. 1). In subfertile mares, the corresponding estimate for embryonic loss between fertilization and Day 50 is 80 per cent, with most embryonic losses occurring before Day 14 in subfertile mares (Fig. 1). The high rate of early embryonic loss in subfertile mares could be related to embryonic defects, oviductal environment, or uterine environment. Oviductal embryos from subfertile mares were less viable than embryos from normal mares; therefore, embryonic defects were important in early embryonic losses in subfertile mares. These defects might be inherent within the embryo or might arise from the early oviductal environment. The uterine environment of subfertile mares was adequate to support normal embryos in early gestation; however, the relationship between the uterine environment and the increased metabolic demands of the conceptus in the late embryonic or early fetal periods requires further study. The uterine environment is also altered in mares with endometritis; therefore, endometritis may also be an important factor in embryonic loss in some mares. Uterine-induced luteolysis, as well as the effect of the pathogen or the resulting inflammation, may lead to embryonic loss. An increased susceptibility of some subfertile mares to endometritis could result in embryonic loss secondary to a postcoital endometritis that persists until the embryo reaches the uterus at Days 5 or 6 postovulation. Although progesterone is critical to embryonic survival, the cause-and-effect relationship between progesterone and spontaneous embryonic loss remains unclear. Reduced progesterone concentrations could be related to endometritis, failure of maternal pregnancy recognition, or luteal insufficiency. Progesterone supplementation may be indicated for some mares, but the value of exogenous progesterone for prevention of spontaneous embryonic loss has not been critically tested. A number of other factors have been associated with embryonic loss in mares.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3044541 TI - Equine artificial insemination. AB - Artificial insemination is an effective technique for improving utilization of the stallion while maintaining normal conception rates in the mare. However, procedures for collection, evaluation, and insemination of semen must be followed carefully to achieve good results. Techniques for preservation of equine semen in the liquid or frozen state could potentially allow for more widespread use of genetically superior stallions. Further acceptance of artificial insemination and the use of cooled or frozen transported semen by breed registries is needed before this will occur. More work is needed to perfect methods of semen preservation, even though semen from some stallions can be cooled or frozen quite successfully at the present time. PMID- 3044542 TI - Equine embryo transfer. AB - Current procedures for collection and transfer of equine embryos are presented. Factors affecting embryo recovery and pregnancy rates after transfer are discussed, and morphologic assessment and development of the embryo are described. PMID- 3044543 TI - Response: "Systems software" letter. PMID- 3044544 TI - Porcelain posterior resin-bonded restorations: current perspectives on esthetic restorative dentistry: Part II. PMID- 3044545 TI - Triazolam an oral sedative for the dental practitioner. PMID- 3044547 TI - Conservative treatment of central giant cell granuloma of the mandible. Report of a case with a 10-year follow-up. PMID- 3044546 TI - Effect of root curettage and sodium hypochlorite treatment on pedicle flap coverage of localized recession. PMID- 3044548 TI - Use of prophylactic antibiotics in oral surgery. PMID- 3044549 TI - Unconventional cancer remedies. PMID- 3044550 TI - Liver transplantation: current concepts. PMID- 3044551 TI - NASA's "space doctor" has the best job on earth. PMID- 3044552 TI - Newfoundland physician does not duck difficult cases. PMID- 3044554 TI - Imipenem: a new carbapenem. Committee on Antimicrobial Agents, Canadian Infectious Disease Society. PMID- 3044553 TI - Acute non-Q-wave myocardial infarction: a distinct clinical entity of increasing importance. AB - Despite the increasing incidence of acute non-Q-wave myocardial infarction, controversy remains regarding its validity as a distinct pathophysiologic physiologic and clinical entity. Review of the data indicates that the controversy is more apparent than real. The pathophysiologic factor discriminating best between non-Q-wave and Q-wave infarction is the incidence rate of total occlusion of the infarct-related artery, approximately 30% in non-Q wave infarction and 80% in Q-wave infarction. Patients with non-Q-wave infarction have a higher incidence of pre-existing angina than patients with Q-wave infarction; they also have lower peak creatine kinase levels, higher ejection fractions and lower wall-motion abnormality scores, which suggests a smaller area of acute infarction damage. However, patients with non-Q-wave infarction have a significantly shorter time to peak creatine kinase level and more heterogeneous ventriculographic and electrocardiographic infarct patterns. The in-hospital death rate is lower in non-Q-wave than in Q-wave infarction (approximately 12% v. 19%). The long-term death rates are similar for the two groups (27% and 23%), but the incidence of subsequent coronary events is higher among patients with non-Q wave infarction; in particular, reinfarction is an important predictor of risk of death. Most of the differences in biologic and clinical variables between the two types of acute infarction can be related to a lower incidence of total occlusion, earlier reperfusion or better collateral supply in non-Q-wave infarction. Further study is needed to better characterize the long-term risk and to define the most appropriate therapies. PMID- 3044555 TI - The Munchausen syndrome: Baron von Munchhausen has taken a bum rap. PMID- 3044556 TI - Dynamics and clinical implications of the nursing home-hospital interface. AB - Nursing homes and hospitals are dynamically interrelated, and there is much that the clinician can do to optimize patient transitions between institutions. This article approaches this topic in three sections: epidemiology of movement between hospitals and nursing homes, impacts of policy changes and financial incentives on patient transfers, and clinical strategies to reduce unnecessary hospitalizations of nursing home patients. PMID- 3044558 TI - Incontinence in the nursing home patient. AB - Prevalent, morbid, and costly, urinary incontinence in the nursing home poses a major problem for patient, caregiver, and administrator alike. In this article, the authors review the pathophysiology of incontinence in the frail elderly, and give guidelines for how to diagnose and treat this common condition. PMID- 3044557 TI - The role of the nursing home medical director. AB - The nursing home medical director faces many overlapping responsibilities. Fostering the development of an interdisciplinary team provides a means of maximizing the influence of the medical director. Setting standards of care and humanizing life in the nursing home are other tasks of the medical director. PMID- 3044559 TI - Assessment and management of dementia in the nursing home. AB - Well over half of the nursing home residents in the United States suffer from dementia, and the absolute number of those affected is expected to increase dramatically. Dementia is a clinical syndrome for which physicians assess patients carefully to search for reversible etiologies and other factors which worsen cognitive function. Behavioral and affective symptoms are common and may be understood as having organic, environmental, psychological, and interpersonal components. Patients with dementia may benefit from judicious use of psychotropics but are often highly vulnerable to adverse effects. Specialized treatment units are showing early promise. PMID- 3044560 TI - Psychiatric disorders among nursing home residents. Depression, anxiety, and paranoia. AB - The number of people in nursing homes is growing at a rapid rate as the population includes greater numbers of elderly Americans. Psychiatric disorders, including depression, anxiety, and paranoia, are commonly seen among this group. Recognition and treatment of these disorders by health care professionals can improve the quality of care and quality of life for elderly nursing home residents. PMID- 3044561 TI - Assessing quality in nursing homes. AB - Quality assurance (QA) in nursing homes requires recognition of differences between acute and long-term care in goals, technology, and pace. This article outlines an approach to nursing home QA that would both generate an information system for a facility to review its performance compared to external regulations, and also allow a proactive stance on the part of nursing home staff. After reviewing existing nursing home regulations, this article discusses three necessary components of QA: defining quality and developing criteria; assessing quality; and correcting deficiencies. It argues that a prompt and clear feedback system to personnel is one key to a proactive QA system that becomes self improving. PMID- 3044562 TI - Decision making by and for nursing home residents. A legal view. AB - The law is concerned with a panoply of issues affecting the care and lives of nursing home residents. This article has outlined one area, that of decision making, which in many respects is the embodiment of and key to all other fundamental resident rights. The doctrine of informed consent applies with full force in the nursing home, both for mentally competent residents and for cognitively impaired residents for whom decision making rights must be exercised through a proxy. Long-term care institutions and professionals are obligated to insure that decisions made by or for residents are made voluntarily, competently, and knowingly. PMID- 3044564 TI - Can psychological assessment address borderline phenomena? AB - Literature about psychological test findings in borderline disorders reflects changes in the meaning of the term and can be confusing when viewed from today's prespective. Most descriptions have referred to a concept of borderline schizophrenia. This paper reviews psychological assessment of borderline patients by means of the Rorschach, by using WAIS/Rorschach patterns, and, post-DSM-III, by using the MMPI. It focuses on the Rorschach's sensitivity to several dimensions relevant to borderline pathology. The concept that borderline disorders can include mild forms of affective and schizophrenic illness was examined by applying a Rorschach content scoring system to a borderline sample. The findings demonstrate Rorschach ability to identify borderline subtypes and offer independent validation of affective and schizotypal subtypes in the borderline realm. PMID- 3044563 TI - Nursing home policies addressing the use or withdrawal of life-sustaining medical treatments. AB - There has been increasing interest in the use of nursing home policies and guidelines to improve decisions about the elective use of life-sustaining treatments. Such policies should be based on a sound and complete understanding of the legal and ethical governance responsibilities of health care facilities. Policies should address both the process of decision making and treatment plan implementation. Nursing home administrators should commission policies and monitor their implementation. PMID- 3044566 TI - Inpatient and day hospital treatment of the borderline: an integrated approach. AB - An integrated treatment approach to the severely ill borderline patient is outlined. Treatment is conceptualized as a process of progression along a continuum of decreasing levels of care. This involves an inpatient, day hospital, and outpatient phase of treatment. During the treatment process, there are many progressions and regressions. The overall goal is to give the patient an increasing sense of separateness, autonomy, and self-reliance. PMID- 3044565 TI - Psychopharmacologic management of patients with borderline personality disorder. AB - This paper reviews recent literature on the psychopharmacologic management of borderline personality disorder (BPD) patients and discusses an approach to drug therapy. Five randomized controlled trials have shown positive, but non-specific effects of antipsychotic drugs on the symptoms suffered by BPD patients. There were too few data on other types of drugs to draw any conclusion. We propose that BPD patients be treated on the basis of being in a state or episode of co existing Axis I disorder. PMID- 3044567 TI - Time-limited, group psychotherapy for borderline patients. AB - Preparation for treatment, group process and time boundaries are presented as components of a treatment model designed to respond to specific problems inherent in the nature of borderline personality disorder. A pilot study of the proposed treatment model is underway and one case illustration from this study is described to illustrate the adaptation to this treatment of a borderline patient with an extensive prior history of individual treatment. PMID- 3044568 TI - [Preconscious and conscious stimulation of erotic imagination]. AB - The stimulation of erotic fantasies through the association of relaxation and erotic conscious or preconscious suggestions has been evaluated. This study was attempted following positive results in the stimulation of fantasmatic activity in alexithymic subjects with a similar procedure. Thirty female subjects, allocated into three groups practiced relaxation daily for two weeks including three sessions with psychological measures. During the second week, erotic suggestions, preconscious for one group and conscious for another one, were added. The third group (control) received only relaxation throughout. Results have shown an increase of sexual arousal and erotic imagery during the sessions with erotic suggestions. Sexual activities and desire increased in the two experimental groups. There was no difference between the effects of the preconscious and conscious suggestions. Possible clinical applications of such a procedure are discussed. PMID- 3044569 TI - Delusional (paranoid) disorders. AB - The group of paranoid or delusional disorders, although not nearly as common as the mood and schizophrenic disorders, may be much more frequent than has usually been thought. DSM-IIIR has made a decisive step in recognizably defining at least one group of them. Interestingly, this change partly came about because the advent of an effective treatment helped to define that group more clearly. Nevertheless, DSM-IIIR's classification is too restrictive, and it was wrong to exclude the diagnosis of paraphrenia. Cases fitting this description will have to be consigned to the category of Psychotic Disorder NOS, which will inevitably be a grab-bag of mixed diagnoses. Also, DSM-IIIR does not emphasize the link between the delusional disorders and paranoid schizophrenia, and the somewhat less well defined overlap with affective disorders, both of which give rise to much diagnostic confusion and inappropriate treatment. Precise history taking and mental status examination and, above all, an up-to-date knowledge of their existence are essential to the recognition and appropriate treatment of the delusional disorders. PMID- 3044570 TI - Clinical neuroscience approaches in psychiatry. AB - Recent advances in the clinical neurosciences have begun to expand and change our understanding of how the brain functions. As further neuroscientific principles are delineated we may gain insights into the underlying pathophysiology of some psychiatric disorders and through this new understanding we may be able to define new therapeutic interventions. Two illustrative examples of neuroscientific research are discussed and reviewed both in terms of the promises and dangers inherent in these new approaches to the mind. PMID- 3044571 TI - Memory and appreciation Henry B.M. Murphy, M.D., Ph.D., 1915-1987. PMID- 3044572 TI - Specification and use of a mouse monoclonal antibody raised against melanosomes for the histopathologic diagnosis of amelanotic malignant melanoma. AB - The positive reactivity and specificity of a mouse monoclonal antibody (MoAb) human melanosome-specific antigen (HMSA) 2 raised against the melanosomal protein with amelanotic malignant melanoma on routine paraffin sections is reported. MoAb HMSA-2 identified cytoplasmic antigen with the following specifications: (1) a sharp heterogeneous expression in melanoma cells (acral lentiginous melanoma [ALM], 11 of 14; superficial spreading melanoma [SSM], 13 of 14; nodular melanoma [NM], one of three; and lentigo maligna melanoma [LMM], zero of two), whereas a diffuse homogeneous expression in cells of benign pigmented melanocytic nevi; and (2) an intense expression on amelanotic melanoma cells as opposed to a weak or negative expression on highly melanotic cells. The positive reactivity of MoAb HMSA-2 with amelanotic melanomas was exemplified by two shave-biopsy specimens of amelanotic subungual and plantar lesions, and by two cases of axillary and cervical amelanotic nodes that were left undiagnosed on routine hematoxylin and eosin (H & E) sections because of lack of melanin pigments. These were found, after diagnosis with MoAb HMSA-2, to possess the regressed primary lesions (both ALM). PMID- 3044573 TI - Endocrine differentiation of gastric adenocarcinoma. The prevalence as evaluated by immunoreactive chromogranin A and its biologic significance. AB - The prevalence of endocrine differentiation of conventional gastric adenocarcinoma was evaluated on the 212 cases (including 62 mucosal carcinomas) of consecutively resected stomach for adenocarcinoma in our hospital using anti chromogranin A (CGA) antibodies. CGA-positive cells were found in 28 of 150 cases (18.7%) as an integral tumor component. In immunocytochemistry and electron microscopic examinations, we could classify these 28 cases into three groups according to the distribution patterns of CGA-positive cells. The first group consisted of 12 cases in which scattered CGA-positive cells were located in neoplastic glands. The second group consisted of six cases of scirrhous carcinoma in which CGA-positive cells were separated by fibrovascular tissue. The third group consisted of ten cases in which the positive cells were present in clusters. No definite correlation was recognized between the appearance of CGA cells and histologic types of predominance. In the analysis of the hormonal substances coexpressed by CGA-positive cells, immunoreactive serotonin (SER) was found most frequently, and somatostatin (SS), gastrin (GAS), glucagon/glicentin (GLU/GLI), and peptide-tyrosine-tyrosine (PYY) like immunoreactivities were found in a few tumor cells. CGA-positive cells occupied limited parts of the tumors in most cases, and they were noticeably more frequent in advanced stage cases. This might explain why endocrine differentiation reflects the dysexpression of the neoplastic stem cells. Furthermore, absence of mitotic figures in this type of cell and negativity of a single colony composed exclusively of CGA cells in metastatic foci suggested that these cells are in a dormant phase and are probably postmitotic. PMID- 3044574 TI - Effect of differentiation-inducing agents on oncogene expression in a chronic myelogenous leukemia cell line. AB - K562 is a Philadelphia (Ph) chromosome-positive chronic myelogenous leukemia (CML) blast crisis cell line representing a pluripotent precursor cell. At the molecular level, K562 cells express high levels of the aberrant bcr-abl product, p210bcr-abl, believed to be critical to the pathogenesis of CML. The authors demonstrate that exposure of K562 cells to hemin causes a state of partial, reversible erythroid maturation, accompanied by a marked decrease in p210bcr-abl. The change in bcr-abl expression may be mediated at the translational level since steady state amounts and enzymatic activity of the bcr-abl protein are reduced whereas bcr-abl mRNA levels are unaltered. The decrease in p210bcr-abl phosphokinase enzymatic activity can be detected within 2 hours after addition of hemin to the culture media, indicating that changes in expression of this oncogene probably occur before or concurrent with differentiation. No change in bcr-abl protein occurred in a CML cell line (KBM-5) which did not undergo differentiation after exposure to hemin, consistent with a direct relationship between altered p210bcr-abl expression and hemin-induced erythroid differentiation. Importantly, the marked diminution in bcr-abl protein was not associated with a disruption in K562 growth rates, indicating that the proliferative capacity of these cells may be independent of the bcr-abl product. In contrast to hemin, cytosine arabinoside (Ara-C) caused terminal erythroid differentiation of K562 cells, characterized by irreversible hemoglobin accumulation and cytostasis; and no change in bcr-abl protein expression was observed. The distinct effects of Ara-C and hemin could reflect the existence of pleiotropic differentiation pathways. Both Ara-C and hemin-exposed cells showed a decrease in c-myc and c-myb transcripts, suggesting that altered levels of these proto-oncogenes may be associated with erythroid maturation, regardless of the rate of cell division. K562 cells provide a useful model for analyzing the interaction between oncogene expression and CML cell growth and differentiation. PMID- 3044575 TI - Psychosocial outcome in a randomized surgical trial for treatment of primary breast cancer. AB - A study of the differences in the psychosocial effects of mastectomy versus segmentectomy was done on a group of women who were in a prospective randomized protocol for treatment of primary breast cancer. Through questionnaires designed for this study and standardized psychologic tests, women with segmentectomies responded as significantly less anxious, less sad, and more in control of their life events than women with mastectomies. The women with segmentectomies had a statistically more positive sexual and body image than those with mastectomies. The trauma of viewing the surgery was much greater in patients with mastectomies. The concern about cancer recurrence was less in the segmentectomy group. The differences in psychosexual adaptation to mastectomy or segmentectomy and the fears of cancer recurrence were significantly better in the segmentectomy group. The adequacies of cancer therapy was the same for both groups in the national study. This study restresses the importance of the segmentectomy option for women with breast cancer in leading to a better quality of life. PMID- 3044577 TI - Rare and polymorphic fragile sites and cancer. PMID- 3044576 TI - Cystic schwannoma mimicking adrenal tumor. AB - The CT and ultrasound findings in a case of cystic schwannoma mimicking a left adrenal tumor are reported. The differential diagnosis of cystic lesions arising in the adrenal region is discussed. PMID- 3044578 TI - Gene expression caused by alkylating agents and cis-diamminedichloroplatinum(II) in Escherichia coli. AB - Previous work has demonstrated heterogeneous effects of methylating agents on induction of DNA damage inducible genes in Escherichia coli. These studies employed E. coli mutants that have fusions of the lac operon to genes induced by treatment with sublethal levels of alkylating agents. These mutants were selected from random insertions of the Mu-dl (Apr lac) phage by screening for induction of beta-galactosidase activity in the presence of methylmethanesulfonate or N-methyl N'-nitro-N-nitrosoguanidine. The current report extends these findings by analyzing gene expression caused by mechlorethamine, chloroethylnitrosoureas and cis-diamminedichloroplatinum(II) (cis-DDP). The results demonstrate heterogeneous effects by these agents on gene expression. While 1-(2-chloroethyl)-3-cyclohexyl 1-nitrosourea induces alkA, other nitrosoureas, mechlorethamine, and cis-DDP do not cause expression of this gene. Further, while all nitrosoureas caused expression of aidC, mechlorethamine and cis-DDP did not. Lastly, cis-DDP caused marked expression of a sulA fusion mutant while not inducing any of the other E. coli fusion mutants. PMID- 3044579 TI - Identification of a tumor-associated target antigen, ATM-1, for a human T-cell clone with activated killer activity and its existence in sera of cancer patients. AB - Three human T-cell clones with activated killer activity (5B5, 5C1, and 7B5) which could lyse various tumor cell lines were established. The cytotoxic activity of these clones was decreased by incubation with anti-CD3 monoclonal antibody, suggesting that they recognized tumor cells by T-cell antigen receptor. A monoclonal antibody which blocked the cytotoxic activity of clone 5B5 was obtained. This antibody (N1977) blocked the binding and cytotoxic activity of clone 5B5 at the target cell level, suggesting that the antigen defined by N1977 antibody, designated as ATM-1, was a target molecule recognized by 5B5 cells. ATM 1 in the conditioned medium of a cancer cell line (NBT-2) and serum from a patient with lung cancer was characterized by following its immunoreactivity. On gel filtration, both the conditioned medium and the serum gave three peaks of ATM 1 immunoreactivity, corresponding to approximate molecular weights of 1,200,000, 700,000, and 120,000, respectively. They were chromatofocused at pH 4.0, 4.8, and 6.5, respectively. The high molecular weight forms were shown to be molecules with the disulfide-linked elementary glycoprotein with ATM-1 immunoreactivity and approximate molecular weight of 120,000. Most of the molecules with ATM-1 immunoreactivity bound to both concanavalin A and wheat germ agglutinin, and their binding activity to the antibodies was lost by treatment at 60 degrees C for 30 min. An assay of ATM-1 level in sera was performed by a sandwich enzyme immunoassay. The following positive percentages were obtained from preliminary clinical studies: breast cancer, 67% (8 of 12 cases); hepatocellular carcinoma, 83% (10 of 12 cases); gastric cancer, 58% (7 of 12 cases); lung cancer, 41% (5 of 12 cases); hematological malignancies, 0% (0 of 9 cases); systemic lupus erythematosus, 0% (0 of 8 cases); rheumatoid arthritis, 0% (0 of 8 cases). PMID- 3044580 TI - Decreased progesterone binding and attenuated progesterone action in cultured human breast carcinoma cells treated with epidermal growth factor. AB - Specific progesterone binding by cultured human breast carcinoma T47D, MCF-7, and ZR75-1 cells was decreased 25-40% by epidermal growth factor (EGF), with a 50% effective dose of 0.1 nM EGF. Studies with the soluble and particulate fractions prepared after homogenization of T47D cells grown in glass roller bottles revealed equivalent EGF-induced decreases in progesterone binding to receptors in both fractions. Equilibrium progesterone binding studies with these soluble and particulate fractions revealed that EGF decreased the receptor number, but had no effect on affinity. With cells grown adherent to plastic dishes, EGF treatment induced a greater decrease in binding to receptors recovered in the particulate fraction, than to receptors recovered in the soluble fraction. The decrease in progesterone binding induced by 20 nM EGF was maximal after 2 min of cellular EGF treatment for receptors recovered in the soluble fraction, but was only half maximal after 15 min for receptors recovered in the particulate fraction. Decreased progesterone binding persisted for at least 8 days in cells cultured with 1 nM EGF. Either insulin or EGF stimulated T47D cell proliferation by two- to threefold with a 50% effective dose of 100 nM for insulin and 0.1 nM for EGF. The progestin, R5020, decreased T47D cell growth by 30% with a 50% effective dose of 1 nM. Either EGF or insulin antagonized the inhibitory effect of R5020 on cell reproduction, but progestins did not antagonize the growth stimulatory response of cells to EGF. Progestins increased the number of EGF receptors within 12 h of their addition to T47D cells, but this response was lost after 6 days. These data show that EGF or progesterone can regulate the receptor number of the other, but for cell reproduction, the effect of EGF is dominant over that of progestins. PMID- 3044581 TI - Amplification of epidermal growth factor receptor gene but no evidence of ras mutations in primary human esophageal cancers. AB - Primary esophageal squamous cell carcinomas from 41 patients were analyzed for the presence of proto-oncogene alterations associated with this malignancy. The occurrence of activating ras gene mutations in 25 tumors was determined using oligomer hybridization of target sequences amplified by polymerase chain reaction. We found no evidence for mutations in codons 12 and 61 of the H-ras, K ras, and N-ras genes, nor in codon 13 of the K-ras and N-ras loci in any of these tumors. The apparent absence of activated ras oncogene in esophageal cancers represents a possible exception to the presence of these mutations found consistently in numerous other types of human malignancies, and is in striking contrast to the 40% prevalence of ras mutations in human colorectal cancers. Southern blot hybridization experiments with DNAs from tumors demonstrated amplification of the epidermal growth factor receptor gene (c-erbB) in two of 25 carcinomas. No amplification of the structurally related c-erbB2 (neu) gene was detected. In three out of 12 carcinomas, the level of epidermal growth factor receptor RNA was significantly higher than in normal esophageal mucosal tissue. Our results suggest that enhanced transcription of the epidermal growth factor receptor gene is associated with the development of some esophageal cancers. PMID- 3044582 TI - Ribonucleotide reductase M1 subunit in cellular proliferation, quiescence, and differentiation. AB - Ribonucleotide reductase catalyzes the first unique, rate-limiting step in DNA synthesis; both its large (M1) and small (M2) subunits are necessary for activity. While direct studies of M2 expression have previously shown a tight correlation with S phase, the kinetic features of M1-expressing cells have remained ill defined. Therefore we have, using immunofluorescence flow cytometry, analyzed changes in whole cell M1 levels and DNA content during various cell cycle and differentiation events. In asynchronous cultures M1 levels are sustained throughout the cell cycle, including G1 phase when M2 levels and ribonucleotide reductase catalytic activity are known to be very low. In contrast M1 is virtually absent from quiescent lymphocytes but is expressed following mitogen stimulation, shortly before S phase cells appear. M1 declines to low levels in "plateau phase" cultures, the major reduction occurring in cells with 2n (G0/G1) DNA content. HL-60 promyelocytic leukemia cells, induced into either myeloid or monocyte-macrophage differentiation, show a similar marked decrease in M1 levels concomitant with the cessation of cell division. We conclude that the M1 subunit of ribonucleotide reductase is constitutively expressed by cycling cells. It is acquired during stimulated transition from G0 to G1 and is lost during exit to G0 or terminal differentiation. This pattern of expression suggests that determination of cellular M1 content may be useful in distinguishing proliferating (including G1) and quiescent (including G0) cells in vivo. PMID- 3044583 TI - Optimal loading of scraped HeLa cells with monoclonal antibodies to the proliferation-associated Mr 120,000 nucleolar antigen. AB - Our laboratory has reported (J. W. Freeman et al., Cancer Res., 48:1244-1251, 1988) a proliferation-associated Mr 120,000 nucleolar antigen (p120), that was found in human tumors but was not detectable in most normal resting tissues and in benign tumors. To study further the function and the localization of this protein, we have investigated various methods of microinjecting p120 monoclonal antibodies into cells. For comparison, we have used a monoclonal antibody to protein C23, a nucleolar protein found in high levels in most cells. To determine optimal conditions for loading of antibodies to nucleolar antigens into mechanically disrupted HeLa cells, we studied the effects of ionic strengths of loading buffers, various antibody concentrations, and optimal time for loading and antibody localization. With ascites fluids in isotonic buffer containing antibodies to nucleolar proteins p120 and C23, a maximum number of cells, 86 and 84%, respectively, were loaded following a 20-min incubation. Hypotonic buffers decreased the percentage of cells loaded (22%); hypertonic buffers reduced cell viability. The optimal concentration of purified antibody yielding a maximum number of loaded cells (81%) was 2.5 mg/ml. Higher concentrations of antibody resulted in residual cytoplasmic staining without increasing the percentage of loaded cells. With antibody concentrations less than 2.5 mg/ml, a linear decrease was noted in the percentage of cells loaded with a decrease in intensity of fluorescence. Following antibody loading, nucleolar fluorescence was observed by 12 h and the intensity increased at 24 h. Localization of the p120 antibody was followed through mitosis where it was perichromosomal and equally divided between the chromosomes at metaphase. A decrease of nucleolar immunofluorescence intensity and percentage of cells labeled were observed in successive cell generations. PMID- 3044584 TI - Blood group-related antigens in human germ cell tumors. AB - With the use of immunohistochemical techniques, seven mouse monoclonal antibodies and the lectin from Ulex europaeus, detecting blood group antigens of the ABH and Lewis systems, have been used to define the distribution of these antigenic structures in germ cell tumors. The reagents used recognize the following blood group antigens: A, B, H, Lewisa, Lewisb, X (Lewisx), Y (Lewisy), and type I precursor antigen. Tumors from 29 patients were studied. Tumors studied consisted of pure embryonal carcinoma for eight patients, pure yolk sac tumor for two patients, embryonal carcinoma plus yolk sac tumor in one patient, and yolk sac tumor plus seminoma in one patient. Also studied were nine classic seminomas and a group of six patients with tumors classified as seminomas that exhibited atypical histological features. One patient had an anaplastic carcinoma arising from the mediastinum which could not be conclusively identified as a germ cell tumor morphologically and was analyzed separately. All embryonal carcinomas and yolk sac tumors exhibited strong positivity for type I precursor structure as detected by the K-21 monoclonal antibody. In marked contrast, there was non staining in classic seminomas but heterogeneous staining in five of six atypical seminomas. The majority of embryonal carcinomas and all yolk sac tumors studied demonstrated strong positivity for blood group antigen H. For seminoma, however, only one of the atypical cases and two of the classic cases (occasional cells) stained for H. Focal expression of the Y antigen was identified in 5 of 17 seminomas and in the majority of embryonal carcinomas and yolk sac tumors. Two yolk sac tumors and two classic seminomas expressed blood group X. The remaining blood group antigens were not expressed by seminomas while they were variably expressed by embryonal carcinoma and yolk sac tumors. These data suggest that K 21 and blood group antigen H may be distinguishing markers of nonseminomatous germ cell tumor versus seminoma. If so, it is possible that the heterogeneous expression of blood group substances in seminomas with atypical histologies is an indication of differentiation towards nonseminomatous germ cell tumor. PMID- 3044585 TI - Prostacyclin and protection of the ischemic myocardium. PMID- 3044586 TI - [Anti-arrhythmia drugs of the '80s: something new with respect for the old]. PMID- 3044587 TI - [Surgical indications in acquired tricuspid valvulopathies]. PMID- 3044588 TI - [Double-blind comparative study of the efficacy of metoprolol and hydrochlorothiazide in the elderly hypertensive. Italian results of an international multicenter study]. PMID- 3044590 TI - Streptococcus mutans counts obtained by a dip-slide method in relation to caries frequency, sucrose intake and flow rate of saliva. AB - The level of Streptococcus mutans in saliva was determined by a dip-slide method in 841 13-year-old children in order to identify children with high caries risk. For each child, the flow rate of saliva was determined. Caries scores were obtained from Public Dental Health records. A sucrose intake score was calculated based on self-reported frequency of intake of six types of sugary products. As S. mutans counts increased, there was a significant trend of increased DMFS and DS scores. No linear correlation was observed between reported intake of sucrose and S. mutans counts, but the children with the highest counts (class 3) tended to have significantly higher sucrose intake than the rest of the children. The flow rate of saliva decreased significantly as S. mutans increased. PMID- 3044589 TI - [Bepridil in stable angina of effort: a double-blind comparison with verapamil]. PMID- 3044591 TI - Dextromethorphan binding sites in the guinea pig brain. AB - 1. Dextromethorphan (DM), a dextrorotatory nonopioid antitussive, binds to specific high-affinity sites in the central nervous system. These sites are distinct from the opioid and other known neurotransmitter receptor sites. Antitussives such as carbetapentane and caramiphen also bind to DM sites with a nanomolar affinity. 2. The anticonvulsant drugs phenytoin and ropizine produce an allosteric enhancement of the binding of [3H]DM to guinea pig brain. DM, carbetapentane, and caramiphen also are efficacious anticonvulsant agents in the rat maximal electroshock seizures test, and DM enhances the anticonvulsant effects of phenytoin (PHT). 3. These results suggest that drugs that bind to the DM sites could be used alone as anticonvulsants or in combination with PHT to lower its effective dose and reduce its side effects. 4. The investigation of the DM binding sites may help to open new approaches for the treatment of convulsive disorders and to explain further some of the molecular mechanisms of neutronal excitability. PMID- 3044593 TI - [A 3-year prospective study of cyclosporine and conventional immunosuppression in kidney transplantation]. PMID- 3044594 TI - [Founding of the Trebon Hospital during World War I]. PMID- 3044592 TI - A new approach to biochemical evaluation of brain dopamine metabolism. AB - 1. Dopaminergic neurotransmission in brain is receiving increased attention because of its known involvement in Parkinson's disease and new methods for the treatment of this disorder and because of hypotheses relating several psychiatric disorders to abnormalities in brain dopaminergic systems. 2. Chemical assessment of brain dopamine metabolism has been attempted by measuring levels of its major metabolite, homovanillic acid (HVA), in cerebrospinal fluid, plasma, or urine. Because HVA is derived in part from dopamine formed in noradrenergic neurons, plasma levels and urinary excretion rates of HVA do not adequately reflect solely metabolism of brain dopamine. 3. Using debrisoquin, the peripheral contributions of HVA to plasma or urinary HVA can be diminished, but the extent of residual HVA formation in noradrenergic neurons is unknown. By measuring the levels of methoxy hydroxyphenylglycol (MHPG) in plasma or of urinary norepinephrine metabolites (total MHPG in monkeys; the sum of total MHPG and vanillyl mandelic acid (VMA) in humans) along with HVA, it is possible to estimate the degree of impairment by debrisoquin of HVA formation from noradrenergic neuronal dopamine and thereby better assess brain dopamine metabolism. 4. This method was applied to a monkey before and after destruction of the nigrostriatal pathway by the administration of MPTP. PMID- 3044597 TI - [Graft function and infectious complications in a permanent graft nephrostomy]. PMID- 3044596 TI - [Chronic pancreatitis. Possibilities of ultrasonographic diagnosis]. PMID- 3044595 TI - [Women in the life of Jan Evangelista Purkinje. I. His mother, his friend and his wife]. PMID- 3044598 TI - [Women in the life of Jan Evangelista Purkinje. II. Women of the Czech national revival period]. PMID- 3044600 TI - Immunocytochemical studies on the LHRH system of the Japanese quail: influence by photoperiod and aspects of sexual differentiation. AB - Immunocytochemistry was used to determine if photoperiod and/or sex have any effect on the pattern of the luteinizing hormone-releasing hormone (LHRH) system in the brain of the Japanese quail. Immunopositive perikarya were found within three major areas of the brain: the rostral paraolfactory lobe, the preoptic, and the septal region. A quantitative analysis of LHRH cell numbers was performed on male and female quail after two photoperiodic treatments: sexually mature birds exposed to 24 weeks of 20 h light: 4 h darkness (20L:4D), and birds with a regressed reproductive system (induced by transfer from a photoregime of 20L:4D to 25 short days of 8L:16D). Two-way analysis of variance showed that short-day males display significantly (p less than 0.05) more immunopositive perikarya (607 +/- 134) than long-day males (291 +/- 114), short-day females (293 +/- 103) or long-day females (330 +/- 92). The density of LHRH-immunoreactive nerve fibres and the intensity of the immunostaining in the median eminence were always greater in long-day sexually mature quail (male and female) than in animals exposed to 25 days of 8L:16D. These results demonstrate that the LHRH system of the quail is influenced by photoperiod and mirrors sexual differentiation. PMID- 3044603 TI - [Advances in research on the serodiagnosis of candidiasis]. PMID- 3044601 TI - Twenty-four hour variations in subcellular structures of rat pancreatic islet B-, A- and D-cells, and of portal plasma glucose and insulin levels. AB - Variations in stomach weights, plasma glucose and insulin levels in the portal vein, and in subcellular structures of rat pancreatic islet B-, A- and D-cells were examined over 24 h. The variation in stomach weights paralleled plasma glucose levels, indicating that the levels may be influenced by intestinal glucose absorption. Plasma insulin levels increased from the late dark to the early light periods, whereas they decreased from the late light to the early dark periods. The variation in plasma insulin levels was in the opposite sense to that in the relative numbers of B-cell granules. The decrease in the relative numbers of A-cell granules occurred between the late dark and early light periods. The relative numbers of D-cell granules decreased before and after the decreases in B and A-cell granules. The variation in D-cell granules appears to correspond to the inhibitory effect of somatostatin. The relative amounts of rough endoplasmic reticulum (rER) in each cell varied in a reciprocal manner to those of B-cell granules. Moreover, the variation in plasma insulin levels coincided with variations in rER of hepatocytes and pancreatic acinar cells. The changes in rER of each cell may correlate with the trophic effect of insulin. PMID- 3044599 TI - Human macrophage maturation and heterogeneity: restricted expression of late differentiation antigens in situ. AB - Terminal maturation of human macrophages is an important step for creation of cell diversity amongst site-specific subpopulations and their functional competence in situ. As monocytes undergo differentiation in vitro, they start to express lineage-restricted antigens specific for differentiation stages beyond the blood monocyte level as detected by monoclonal antibodies of the MAX series. We have analyzed the expression of MAX.1, MAX.2, MAX.3 and MAX.11 on exudate-type macrophages from pleural and peritoneal cavity and the alveolar space, as well as on resident and activated tissue macrophages in cryostat sections of spleen, lymph node, tonsil, liver, gut mucosa, skin, placenta, kidney and bone. It was found that "free" macrophages in serous cavities expressed MAX antigens in a heterogenous pattern, whereas none of the organ-specific tissue macrophages subsets did so (with the exception being the weak label of MAX.2 on Kupffer cells). Only during allograft rejection were infiltrating macrophages found to express MAX antigens but not at sites of "non-specific" inflammation or granuloma formation. However, Cyclosporin A treatment seems to suppress the induction of MAX antigen expression on intragraft macrophages. In addition, freshly harvested MAX-negative exudate macrophages converted to the complete Max+ phenotype on further cultivation. Isolated Kupffer cells were able only to express the MAX.2 antigen in culture but still did not react with the MAX.1 and MAX.3 monoclonal antibodies. Some MAX antigens are co-expressed on glomerular mesangial cells, dendritic reticulum cells and placental cells (MAX.1/.11) as well as on capillary endothelium within tissues of active immune response (MAX.2).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3044604 TI - [Serotypes of enteroinvasive Escherichia coli in China]. AB - results of identification and serotyping of enteroinvasive escherichia in china from 13 provinces in China in 1984-85 were reproted. PMID- 3044602 TI - Colocalization of glucagon-like peptide and glucagon immunoreactivities in pancreatic islets and intestine of salmonids. AB - Pancreatic islets of salmon contain at least two peptides of the glucagon family: 29-amino acid glucagon and 31-amino acid glucagon-like peptide (GLP). Both peptides were recently isolated from the pancreatic islets of coho salmon and sequenced (Plisetskaya et al. 1986). Antibodies generated against these two peptides and against human glucagon were used as immunocytochemical probes to investigate whether glucagon and GLP are processed in the same, or in different cell types in the pancreatic islets and the gut of salmon. Two salmonid species, rainbow trout and coho salmon, were studied. All islet A-cells in the two species were immunoreactive toward both anti-salmon (s)-glucagon and anti-s-GLP. Similar colocalization of glucagon and GLP immunoreactivites was found in open-type endocrine cells in mucosae of the small intestine (including the pyloric coecae) and the large intestine close to the vent of rainbow trout. None of the antibodies stained mucosal cells of the body of the stomach. These results suggest that in the pancreas and the gut of salmonid fish the same cells produce both glucagon and GLP. These peptides are most likely the products of a single gene coding for the preproglucagon sequence. PMID- 3044605 TI - Transforming growth factor alpha. PMID- 3044606 TI - Structure of the beta subunit of translational initiation factor eIF-2. AB - A human liver cDNA encoding the beta subunit of protein synthesis initiation factor 2 (eIF-2) was isolated and sequenced. The 1416 bp cDNA encodes a protein of 333 amino acids (38,404 daltons) with characteristics that resemble authentic purified eIF-2 beta. De novo synthesized eIF-2 beta from cDNA transcripts incorporates into endogenous rabbit eIF-2 complexes. The protein possesses putative GTP-binding sites, a zinc finger motif, and a highly charged N-terminal region composed of three basic polylysine blocks separated by acidic domains. The polylysine blocks and the zinc finger motif suggest that eIF-2 beta interacts with RNA. A yeast protein, Sui3, isolated as an extragenic suppressor of his4 initiation codon mutations, exhibits extensive sequence identity with human eIF-2 beta, especially in the polylysine and zinc finger domains, thereby reinforcing the view that these elements are important for function. PMID- 3044607 TI - GAL4 derivatives function alone and synergistically with mammalian activators in vitro. AB - We show that derivatives of the yeast transcriptional activator GAL4, synthesized in and purified from E. coli, stimulate transcription of a mammalian gene (the adenovirus E4 gene) in a HeLa cell nuclear extract. Stimulation depended upon GAL4 binding sites inserted in the template. When the GAL4 sites were placed immediately upstream of the E4 TATA box, GAL4 stimulated efficiently. When the GAL4 sites were further upstream from TATA, however, efficient stimulation by GAL4 required, in addition, a site for a mammalian transcriptional activator immediately upstream of TATA. Under these conditions, the GAL4 derivatives functioned synergistically with the proximally bound activator. Previous experiments have defined two "activating regions" in GAL4, and our current experiments define a third, whose function is observed in vitro but not in vivo. PMID- 3044609 TI - DNA dynamic flexibility and protein recognition: differential stimulation by bacterial histone-like protein HU. AB - The abundant E. coli "histone-like" protein HU is shown to be a differential effector of DNA recognition by three diverse control proteins. DNA recognition by lac repressor and catabolite activator protein is greatly stimulated, while specific aroH DNA recognition by trp repressor is inhibited. BaCl2, an agent previously shown to promote DNA bending, mimics the HU effect to give the same qualitative differential stimulation spectrum. The HU activation involves cooperativity, further suggesting that the various DNA bends and distortions induced during assembly of higher order HU:DNA structures are important for the HU stimulation. Thus, E. coli chromosomal DNA regulation is likely strongly influenced by HU protein that may promote a variety of alternative DNA structures that either facilitate or inhibit specific recognition by diverse control proteins. PMID- 3044608 TI - Mechanism of action of a yeast activator: direct effect of GAL4 derivatives on mammalian TFIID-promoter interactions. AB - We have analyzed interactions between the mammalian TATA factor (TFIID) and derivatives of the yeast activator GAL4. The interaction of the TATA factor on the adenovirus E4 promoter with GAL4 binding sites adjacent to the TATA site was qualitatively altered in response to GAL4 binding. Alterations in the TFIID interactions were observed with two GAL4 derivatives that stimulated hybrid E4 promoter activity in vitro but not with a third derivative that bound to DNA but showed no activation. These results indicate that TFIID is a direct target for a GAL4 activation domain and suggest a simple general model for the activation mechanism. PMID- 3044610 TI - SPA1: a gene important for chromosome segregation and other mitotic functions in S. cerevisiae. AB - Human autoantibodies that recognize the spindle poles of mammals, plants, and insects were found to recognize two antigens in yeast. One of these proteins, called SPA1 (for Spindle Pole Antigen), is antigenically related to the spindle poles of a diverse set of organisms. The gene encoding SPA1 was cloned by immunoscreening a lambda gt11 yeast genomic DNA expression library with autoantibody probes. Mutational analysis of the SPA1 gene demonstrates that it is important for cell growth, chromosome segregation, and other cellular processes; spa1 mutants are viable but grow poorly at 30 degrees C, missegregate chromosomes at an increased frequency, and often contain deformed spindles. A significant fraction of spa1 mutant cells contain two or more nuclei, and others contain none; these abnormal cells may arise through a nuclear migration defect. Thus SPA1 represents a new fidelity gene that is important for chromosome segregation and other mitotic functions. PMID- 3044611 TI - A novel nucleotide excision repair for the conversion of an A/G mismatch to C/G base pair in E. coli. AB - A protein that binds specifically to A/G mismatches has been detected in E. coli by a gel electrophoresis DNA binding assay. A specific endonuclease is associated with the A/G mismatch-binding protein through two chromatographic steps. The endonuclease is specific for A/G-containing DNA fragments and has no cleavage activity on DNA containing the other seven possible mispairs or homoduplex DNA. The endonuclease simultaneously makes incisions at the first phosphodiester bond 3' to and the second phosphodiester bond 5' to the dA of the A/G mismatch. No incision site was detected on the other strand. These results are consistent with the unidirectional A to C conversion and short repair tract of a novel dam- and mutHLS-independent A/G repair pathway we have recently described. A nucleotide excision repair model is proposed for the conversion of an A/G mismatch to a C/G base pair. PMID- 3044612 TI - Isolation of the gene encoding the S. cerevisiae heat shock transcription factor. AB - The yeast heat shock transcription factor gene, HSF1, has been isolated from an S. cerevisiae genomic expression library (in lambda gt11). The sequenced gene encodes an 833 amino acid protein having a mass of 93,218 daltons. Expression of specific DNA-binding activity in E. coli and of transcriptional activity in yeast confirmed the identity of the cloned gene. Southern analysis and gene-disruption experiments indicate that the heat shock transcription factor is encoded by a single-copy, essential gene. The DNA-binding domain was localized to a 118 amino acid region in the amino-terminal third of the protein. Inspection of the DNA binding domain reveals no resemblance to any currently known secondary structural motifs implicated in DNA recognition and binding. PMID- 3044613 TI - Yeast heat shock factor is an essential DNA-binding protein that exhibits temperature-dependent phosphorylation. AB - Heat shock promoters contain one or more binding sites for a specific heat shock factor (HSF). We report the cloning and sequence of the gene encoding yeast HSF, and demonstrate that HSF is required for growth at normal temperatures (15 degrees C-30 degrees C). The activity of a promoter containing a synthetic HSF binding site varies over a 200-fold range between 15 degrees C and 39 degrees C (heat shock). This change in activity is accompanied by multiple changes in the phosphorylation state of HSF, but all forms of HSF are able to bind DNA. We propose that the expression of heat shock genes in yeast is modulated by phosphorylation of DNA-bound HSF, and that this leads to a more efficient interaction of the factor with other components of the transcriptional machinery. PMID- 3044614 TI - Two mRNAs that differ by two nontemplated nucleotides encode the amino coterminal proteins P and V of the paramyxovirus SV5. AB - The "P" gene of the paramyxovirus SV5 encodes two known proteins, P (Mr approximately equal to 44,000) and V (Mr approximately equal to 24,000). The complete nucleotide sequence of the "P" gene has been obtained and is found to contain two open reading frames, neither of which is large enough to encode the P protein. We have shown that the P and V proteins are translated from two mRNAs that differ by the presence of two nontemplated G residues in the P mRNA. These two additional nucleotides convert the two open reading frames to one of 392 amino acids. The P and V proteins are amino coterminal and have 164 amino acids in common. The unique C terminus of V consists of a cysteine-rich region that resembles a cysteine-rich metal binding domain. An open reading frame that contains this cysteine-rich region exists in all other paramyxovirus "P" gene sequences examined, which suggests that it may have important biological significance. PMID- 3044615 TI - Cyclic changes in the differentiation of lymphoid cells in reptiles. PMID- 3044616 TI - Biomechanics of the foot and ankle during gait. AB - Running is a very popular activity, whether for competition or fitness. Breakdown injuries related to training errors, shoe wear, or change in intensity are frequently seen by the sports medicine physician. In order to understand and treat the pathologic situation, a fundamental understanding of the biomechanics of walking and running is essential. The treating physician must appreciate the distinct differences between the walking and running gait. These differences transcend a simple increase in speed of gait and include distinct changes in joint range of motion and electromyographic activity. Armed with this knowledge, the physician treating a breakdown injury can work to a solution based on scientific understanding rather than anecdotal information. PMID- 3044617 TI - Tendon injuries about the ankle in athletes. AB - Tendinous injuries about the foot and ankle are often the result of overloading and repetitive microtrauma stemming from training errors, poor technique, and the use of inappropriate surfaces or equipment. Other factors that must be considered include anatomic malalignment and muscle imbalance. Injuries involving tendons include inflammation, subluxation, and rupture. A correct and timely diagnosis should assist athletes, trainers, coaches, and physicians in preventing and treating these disorders. PMID- 3044618 TI - Plantar fasciitis. Mechanics and pathomechanics of treatment. AB - An excessive amount and/or a prolonged duration of pronation is the most common mechanical cause of structural strain resulting in plantar fasciitis. Temporary relief of pain can be achieved by customary antiinflammatory drugs or therapy; long-term relief is achieved by adequate remedy of the aggravating pronation factors. A semirigid, custom-molded orthosis reduces excessive plantar fascial strain by supporting the first metatarsal bone and by controlling calcaneal position when in conjunction with a firm posterior counter shoe. A clinical environment with physician and orthotist together allows ideal evaluation and treatment of patients. PMID- 3044619 TI - Magnetic resonance imaging of the ankle and foot. AB - MRI has gained wide acceptance in evaluating the musculoskeletal system because of its wide array of advantages, including its superior soft-tissue contrast, noninvasiveness, direct multiplanar imaging, and lack of ionizing radiation. Its high-resolution images allow excellent visualization of bones, muscles, ligaments, tendons, and cartilaginous elements. MRI's greater sensitivity in detecting effusions and focal areas of pathology over other currently available imaging modalities make it the method of choice in many instances when imaging the foot and ankle. PMID- 3044620 TI - Rehabilitation of the injured ankle. AB - Stimulation of damaged sense receptors is essential to proper rehabilitation of the injured ankle. By performing exercises that induce movement and change tone and intra-articular pressure, especially in the weightbearing role, the ankle joint's periarticular structures are more likely to heal with functional strength and stability. PMID- 3044621 TI - Hallux rigidus. AB - This article discusses the pathophysiology in the development of the hallux rigidus deformity. The clinical and radiographic findings are discussed and identified. Various modalities of conservative and surgical care are discussed. PMID- 3044622 TI - Sesamoid foot problems in the athlete. AB - Problems of the sesamoid bones of the metatarsophalangeal joint of the great toe are critical in maintaining full ability to compete or to engage in recreation- in athletes of any age. The sesamoids are subject to the same post-traumatic, acute, and chronic changes as any other bone. These unique and deceptively important injuries demand a high index of suspicion, a thorough diagnostic effort, and a guided long-range treatment program, including follow-up to normalized athletics, particularly in the running-based sports. PMID- 3044623 TI - The surgical treatment of hallux valgus using the modified Z-osteotomy. AB - Numerous therapeutic options are available in treating the painful bunion. The procedure that accomplishes the desired results and correction must be carefully matched with the underlying pathologic deformity. For the majority of patients, in the author's hands, the Silver Triad (eminence resection, lateral release, and medial plication) combined with the Z-1st metatarsal osteotomy is the ideal procedure. PMID- 3044624 TI - [Gamete fallopian tube transfer in conjunction with microsurgery of the adnexa]. PMID- 3044626 TI - [Rumen bacteria and their ureolytic activity during nitrogen utilization in ruminants]. PMID- 3044627 TI - Kinetic study on the inhibition of type Ib penicillinase by imipenem and thienamycin. PMID- 3044625 TI - [Reactivity of the pulmonary vessels in hypoxic pulmonary hypertension]. PMID- 3044628 TI - Synthesis of 15-cis-(4-n-propylcyclohexyl)-16,17,18,19,20-pentanor-9- deoxy-6,9 alpha-nitriloprostaglandin F1 methyl ester (OP-2507), a novel anti-cerebral ischemic agent. PMID- 3044629 TI - Proteolytic enzyme sensors using an ion-sensitive field effect transistor. PMID- 3044630 TI - [Study on the trace elements in traditional Chinese drugs over the last several years]. PMID- 3044631 TI - Urinary protein and enzyme excretion in patients receiving chemotherapy with the cis-platinum analogs carboplatin (CBDCA, JM8) and iproplatin (CHIP, JM9). AB - Urinary protein and enzyme excretion was measured in 33 patients with solid tumours receiving chemotherapy with the cis-platinum analogs carboplatin (JM8, CBDCA) and iproplatin (JM9, CHIP). The patients were given up to six courses of the drugs at 4-week intervals, and serial urine samples were collected weekly for periods up to 28 weeks. Overall there was no significant increase in the alkaline phosphatase (ALP), lactate dehydrogenase (LD), and N-acetyl glucosaminidase (NAG) excretion of the first posttreatment samples compared with the pretreatment samples. During the course of treatment there were transient increases in all three enzymes, some quite marked. There was no consistent increase in urinary protein or enzyme excretion during the period of treatment, suggesting that there was no cumulative nephrotoxicity. There was no change in creatinine clearance or urinary beta 2-microglobulin content. Iproplatin appeared marginally more toxic on the basis of elevated NAG and ALP during the second half of the treatment periods compared with the first (P less than 0.01 and less than 0.025, respectively). PMID- 3044632 TI - An autoradiographic study of the intrarenal localisation and retention of cisplatin, iproplatin and paraplatin. AB - The intrarenal localisation of platinum following the intravenous administration of platinum-195m-labelled cisplatin, iproplatin and paraplatin was studied using autoradiography. Following injection of cisplatin, platinum was distributed throughout the kidney even up to 14 days after treatment. In the case of iproplatin and paraplatin rapid platinum clearance was noted from the glomeruli, blood vessels and renal medulla within 2 h of administration. Relative cortical and medullary platinum radionuclide concentrations for all three agents were determined by Chalkley grid analysis. This showed greater relative concentrations of platinum in the cortex at increasing times following iproplatin and paraplatin compared with cisplatin. It is suggested that the lower renal toxicity of iproplatin and paraplatin than of cisplatin may be due to reduced platinum retention within the pars recta. PMID- 3044633 TI - Intensive chemotherapy with high doses of BCNU, etoposide, cytosine arabinoside, and melphalan (BEAM) followed by autologous bone marrow transplantation: toxicity and antitumor activity in 26 patients with poor-risk malignancies. AB - Twenty-six patients (median age 33 years) with poor-risk malignancies were treated with high-dose combination chemotherapy associating BCNU-etoposide cytosine arabinoside and melphalan (BEAM) followed by autologous bone marrow transplantation (ABMT). Twenty-one patients had malignant lymphomas, three, acute lymphoblastic leukemia (ALL), and two, malignant thymomas. Eleven patients (group 1) were not in complete remission (CR) at the time of BEAM, and fifteen patients (group 2) were in CR. Hematological recovery occurred in all patients. The duration of aplasia and the non-hematological toxicities were similar in both groups. Ten of the eleven patients (group 1) evaluable for response achieved CR and one achieved partial remission (PR). Five patients relapsed, and five are in continuous CR with a short follow-up (median 8 months). Among the fifteen patients in CR at the time of BEAM (group 2), four patients relapsed and ten patients are in unmaintained continuous CR with a median follow-up of 15 months (one patient died in CR). The disease-free survival is 53%, with 29% for patients receiving BEAM while in relapse (group 1) and 65% for patients receiving BEAM while in CR (group 2). These data indicate that BEAM followed by ABMT can produce a high antitumor response with an acceptable toxicity in patients with poor-risk malignancies. PMID- 3044634 TI - The disposition of carboplatin in ovarian cancer patients. AB - Carboplatin was given as a 30-min infusion to 11 ovarian cancer patients at doses of 170-500 mg/m2. The ages, weights, and creatinine clearances (Clcr) ranged from 44 to 75 years, from 44 to 74 kg, and from 32 to 101 ml/min, respectively. Plasma, plasma ultrafiltrate (PU), and urine samples were obtained at appropriate times for 96 h and were analyzed for platinum. The PU and urine were also analyzed for the parent compound by HPLC. In patients with a Clcr of about 60 ml/min or greater, carboplatin decayed biexponentially with a mean t1/2 alpha of 1.6 h and a t1/2 beta of 3.0 h. The mean (+/- SD) residence time, total body clearance, and apparent volume of distribution were 3.5 +/- 0.4 h, 4.4 +/- 0.85 l/h, and 16 +/- 3 l, respectively. Cmax and AUCinf values increased linearly with dose, and the latter values correlated better with the dose in mg than in mg/m2. No significant quantities of free, ultrafilterable, platinum-containing species other than the parent compound were found in plasma, but platinum from carboplatin became protein-bound and was slowly eliminated with a minimal t1/2 of 5 days. The major route of elimination was excretion via the kidneys. Patients with a Clcr of 60 ml/min or greater excreted 70% of the dose as the parent compound in the urine, with most of this occurring within 12-16 h. All of the platinum in 24-h urine was carboplatin, and only 2%-3% of the dosed platinum was excreted from 48 to 96 h. Patients with a Clcr of less than about 60 ml/min exhibited dose-disproportional increases in AUCinf and MRT values. The latter were inversely related to Clcr (r = -0.98). Over a dose range of 300-500 mg/m2, carboplatin exhibited linear, dose-independent pharmacokinetics in patients with a Clcr of about 60 ml/min or greater, but dose reductions are necessary for patients with mild renal failure. PMID- 3044635 TI - Continuous vincristine infusion as part of a high dose chemoradiotherapy regimen: drug kinetics and toxicity. AB - A 5-day continuous infusion of vincristine (VCR; total dose 4 mg/m2) has been given as part of a high-dose chemoradiotherapy regimen with bone marrow transplantation. Evidence of neurotoxicity, such as weakness, paraesthesia and intestinal hypomotility, was evaluated prospectively in nine patients. Five patients had advanced neuroblastoma and four, relapsed sarcomas, and all had responded to initial conventional-dose therapy. VCR was combined with high-dose melphalan (180 mg/m2) and fractionated total-body irradiation. Plasma concentrations of VCR were measured by radioimmunoassay during and up to 24 h after the infusion. Serum and urine electrolytes and liver function tests were measured during VCR treatment and at regular intervals thereafter. VCR concentration at 1 h ranged from 1.8 to 10.9 (median 6.6) ng/ml, and a steady state was achieved by 13-30 h (median 16 h). Levels above 1 ng/ml were maintained throughout the 5-day period with a mean steady-state concentration of 1.7 ng/ml (range 1.3-2.15). After cessation of the infusion, serum concentrations fell to below 0.25 ng/ml within 24 h. Abdominal pain occurred in one patient, but neither constipation nor ileus was seen. In two patients severe muscle pain occurred in the lower limbs towards the end of the infusion. Significant electrolyte problems did not occur and, in particular, there was no evidence of inappropriate ADH secretion. Transient increases in liver enzymes were common but bilirubin was not elevated during the period of monitoring. This regimen allows a two-fold escalation in the dose of VCR to be administered, producing sustained high serum drug levels without major toxicity. PMID- 3044636 TI - Mutation site specificity of N-nitroso-N-methyl-N-alpha-acetoxybenzylamine: a model derivative of an esophageal carcinogen. AB - A total of 171 mutations induced by N-nitroso-N-methyl-N-alpha-acetoxybenzylamine within the first 180 base pairs of the lacI gene of Escherichia coli were characterized at the DNA sequence level. Consistent with the methylating ability of this compound and the predicted mutagenic specificity of the O6-methylguanine lesion, all but two of these mutations were G:C----A:T transitions. An analysis of neighboring sequences revealed the same 5' flanking sequence influence on mutability reported for other SN1-type direct-acting alkylating agents. G:C--- A:T transitions were found to be six times more likely to occur at G:C base pairs at which the guanine residues were flanked (5') by a purine than at those preceded by a pyrimidine. This mutagenic and site specificity appeared to be independent of the dose and likely reflects the behaviour of the activated parent carcinogen, N-nitroso-N-methyl-N-benzylamine in the esophagus. PMID- 3044637 TI - Mutational specificity of N-methyl-N-nitrosourea in the lacI gene of Escherichia coli. AB - The mutational specificity of the carcinogenic alkylating agent N-methyl-N nitrosourea (MNU) was investigated through the DNA sequence characterisation of 104 lacI-d mutations induced by this agent in the lacI gene of Escherichia coli. The vast majority (95%) of MNU-induced mutations were G:C----A:T transitions. An analysis of neighbouring base sequence revealed that this transition was almost 10 times more likely to occur if the mutated guanine residue was preceded (5') by a purine (R) rather than a pyrimidine (Y); apart from this 5'-R-G-3' influence no other site specificity of mutation was observed. This 5'-flanking base influence on mutability has also been observed in this system with other mechanistically similar alkylating agents. PMID- 3044638 TI - Morphological transformation of immortalized hamster dermal fibroblasts following treatment with simple alkylating carcinogens. AB - We have examined the mechanism of transformation of a line of immortalized hamster dermal fibroblasts (4DH2 cells) following treatment with the simple alkylating agents, N-methyl-N-nitrosourea (MNU), N-ethyl-N-nitrosourea (ENU) and dimethyl sulphate (DMS). Treatment of 4DH2 cells with the potent point mutagens MNU and ENU gave rise to a spectrum of foci of different sizes, including progressively growing large foci and compact small foci. In contrast, treatment with the weak point mutagen DMS produced mostly large foci. The ability of cell lines derived from morphologically transformed foci to grow in soft agar in general reflects their original size. Thus most cell lines derived from large foci grew in soft agar while most lines derived from small foci did not. Transfection of cellular DNAs into the parent 4DH2 cell line and into NIH3T3 mouse fibroblasts has revealed the presence of dominantly acting transforming genes in the chemically transformed cell lines. Thus DNA from five of six cell lines derived by culturing large foci and from one of three cell lines derived by culturing small foci induced efficient morphological transformation of the recipient cells. Southern analyses of DNA from primary and secondary transfectants showed that several of the transforming genes transferred in these experiments were not closely related to H-ras, K-ras or N-ras. PMID- 3044639 TI - Insights into coronary artery disease gained from metabolic imaging. AB - Positron emission tomography offers the possibility of evaluating and quantifying regional myocardial blood flow and metabolism. Used in patients with coronary artery disease, positron emission tomography has demonstrated sustained metabolic activity in regions with reduced blood flow and impaired contractile function, and it thereby enables differentiation between viable myocardium and myocardium that has succumbed to necrosis and scar formation. Viable myocardial regions identified by metabolic rather than functional or blood-flow criteria are frequently observed in patients after an acute coronary event and in patients with stable coronary artery disease. Positron emission tomography reflects either acute myocardial ischemia, "hibernation," as well as "myocardial stunning." Findings from metabolic imaging have proved useful in characterizing more accurately coronary artery disease and its functional consequences. These findings have been found equally useful for clinical management. PMID- 3044640 TI - Carotid endarterectomy based on duplex scanning without preoperative arteriography. AB - Between July 1985 and September 1987, 25 patients underwent 26 carotid endarterectomies based on an abnormal duplex scan (B-mode ultrasonography and pulsed-Doppler sound spectral analysis) indicative of severe stenosis or ulceration. Arteriography was not performed because of severe unstable angina requiring coronary artery bypass grafting (23 patients) or patient preference (two). Twelve patients were symptomatic, and 13 were asymptomatic but had severe (greater than or equal to 75%) bilateral or unilateral carotid artery stenosis. Operative and pathological analyses confirmed the duplex-scan findings in all 25 cases. All 25 patients survived the operation. One patient had a transient ipsilateral neurological deficit, and one had a permanent contralateral neurological deficit. Five patients died of ventricular arrhythmias within 30 days of operation. Duplex scanning is an accurate method for determining the presence of clinically and hemodynamically significant carotid arterial occlusive disease. Duplex scanning also serves as an alternative method for evaluating patients for whom carotid arteriography may be associated with significant risk. PMID- 3044641 TI - Telemetric detection of cardiac allograft rejection. Correlation of electrophysiological, histological, and biochemical changes during unmodified rejection. AB - Intra-abdominal mismatched heterotopic cardiac allograft transplantation without immunosuppression was performed in eight pigs. Postoperatively, daily electrophysiological studies were carried out either with exteriorized temporary epicardial pacing wires (n = 4) or by a telemetrically controlled implanted pacemaker connected to permanent epicardial pacing leads (n = 4). Electrophysiological studies data were correlated with histopathologic and biochemical findings from daily myocardial biopsies. Electrophysiological studies revealed no significant alteration of sinus or atrioventricular node function, refractoriness, or ventricular pacing threshold. However, ventricular voltage amplitude, measured through the electrodes, decreased steadily with time in all donor hearts and was significantly correlated with histopathologic rejection grade (p less than 0.001) and with adenosine 5'-triphosphate (ATP) depletion (p less than 0.001). Ventricular voltage amplitude less than 75% of baseline occurred 4.5 +/- 1.5 days after transplantation, and this decreased voltage amplitude coincided with a moderate to severe (Grade 2 or 3) histological rejection pattern with a sensitivity of 89% (17 of 19) and a specificity of 77% (17 of 22). Similar changes in voltage amplitude were not found in control hearts. Myocardial tissue ATP values fell significantly from control values with early (Grade 1) rejection (p less than 0.05). Evidence for oxygen free radical injury was indicated by a rise in conjugated dienes of free fatty acids; this increase in diene level occurred 4.3 +/- 1.2 days postoperatively and then regressed during the terminal stages.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3044642 TI - Urgent surgical reperfusion in acute evolving myocardial infarction. A randomized controlled study. AB - To assess the benefit of immediate surgical reperfusion over conventional medical treatment during the first acute evolving transmural myocardial infarction, 68 patients presenting within 4 hours of onset of chest pain were randomized into a medical group and a surgical group. Both groups were comparable for age, sex, coronary risk factors, location of infarct, and coronary anatomy. Radionuclide global ejection fraction at 48 hours after admission was 45 +/- 15% for the medical group versus 50 +/- 15% for the surgical group; at 3 months, ejection fraction values were 51 +/- 13% and 51 +/- 13%, respectively (p = NS). The average radionuclide wall-motion scores (normal, 3) at 3 months were 2 +/- 0.6 for the medical group and 2 +/- 0.7 for the surgical group. There were three (8.8%) early and four (11.7%) late deaths in the medical group and only one (2.9%) early death in the surgical group. Urgent surgical reperfusion in acute evolving myocardial infarction is a safe and effective procedure that appears to reduce early and late mortality but does not appear to improve left ventricular function. PMID- 3044644 TI - 1987 CSCC-MedChem award recipient. PMID- 3044643 TI - Cleveland Clinic Quarterly 1961: Cushing's syndrome. By E. Perry McCullagh. PMID- 3044645 TI - Pharmacogenetics of methylation: relationship to drug metabolism. AB - Pharmacogenetics is the study of inherited variation in drug response. Genetic differences in drug metabolism are the most common causes for inherited variations in drug response or adverse reactions to medications. Methyl conjugation is an important pathway in the biotransformation of many drugs. Experiments performed during the past decade showed that individual variations in the activities of enzymes that catalyze S-methylation, O-methylation and N methylation are under genetic control in human tissue. These inherited variations are responsible for individual differences in metabolism, effect, and toxicity of drugs that undergo methyl conjugation. The approach used to study the pharmacogenetics of methylation may also be applicable to the study of inherited variations in other pathways of drug metabolism. PMID- 3044646 TI - Serum isoforms of creatine kinase isoenzymes. AB - The CK-2 and CK-3 isoenzymes of human serum creatine kinase (CK) can be further subdivided into five isoforms (subforms derived from the same isoenzyme). Three are derived from CK-3 and two from CK-2. The formation of these isoforms is a postsynthetic phenomenon brought about by a serum carboxypeptidase that acts on the M monomer of the enzyme. Sera from healthy subjects contain CK-3(1) as the dominant isoform with lesser amounts of CK-3(2) and CK-3(3). Following damage of muscle tissue, the serum isoform distribution changes as a result of the increased release of CK enzyme. This provides more diagnostic information concerning acute myocardial infarction and other muscle diseases than is available from routine CK isoenzyme analysis. PMID- 3044647 TI - Determination of cholic acid and chenodeoxycholic acid pool sizes and fractional turnover rates by means of stable isotope dilution technique, making use of deuterated cholic acid and chenodeoxycholic acid. AB - A procedure is described for the simultaneous determination of cholic acid and chenodeoxycholic acid pool sizes and fractional turnover rates. After oral administration of known amounts of 11,12-dideuterated chenodeoxycholic acid and 2,2,4,4-tetradeuterated cholic acid, the ratios of chenodeoxycholic acid D2/chenodeoxycholic acid and cholic acid-D4/cholic acid are measured in consecutive serum samples, after which fractional turnover rates and pool sizes of chenodeoxycholic acid and cholic acid are determined arithmetically. In 7 healthy volunteers pool sizes for chenodeoxycholic acid and cholic acid were 22.9 +/- 7.8 and 24.1 +/- 11.7 mumol/kg, respectively. The corresponding values for the fractional turnover rates were 0.23 +/- 0.10 and 0.29 +/- 0.12/day. After oral administration of the labelled bile acids in capsule, the obtained pool sizes were significantly higher than after administration in a bicarbonate solution. Bile acid kinetics were also performed in a patient suffering from a cholesterol synthesis deficiency and in a patient very likely suffering from a bile acid synthesis deficiency. Furthermore, the kinetics of the intestinal absorption and hepatic clearance of unconjugated bile acids have been investigated in 2 healthy subjects. PMID- 3044648 TI - Effect of Cyclosporin A on rat thymus: time course analysis by immunoperoxidase technique and flow cytofluorometry. AB - The effect of Cyclosporin A (CyA) administration 20 mg/kg body weight i.p. for 3 or 7 days on rat lymphoid tissues, especially on the thymus, and the recovery after stopping CyA treatment were investigated by both immunoperoxidase technique and flow cytofluorometry using monoclonal antibodies against rat lymphocytes: OX6, OX7, OX8, OX18 and W3/25. The marked reduction of thymic medulla with CyA treatment was clearly demonstrated by staining with OX18. This change was maximal 7 to 10 days after the start of CyA administration. The obvious restitution of the thymic medulla occurred about 7 days after stopping CyA and was almost completed within 14 days. Flow-cytofluorometric analysis of the thymus showed that the percentages of positive cells labelled with OX7, OX8, OX18 and W3/25 appeared to be not changed except for OX18 during and after CyA treatment. However, the expression of each antigen per cell changed in the amount; the peak of fluorescence intensity of OX7+ cells showed a temporary shift to the right during CyA treatment. Bright positive cell populations for each OX8 and W3/25 increased relatively during CyA treatment, and reverted to the normal levels soon after stopping the CyA treatment. On the other hand, bright OX18+ cells decreased with CyA treatment, but this change recovered gradually after stopping the CyA treatment. Treatment with CyA gave no significant changes in the flow cytofluorometric analyses for these antibodies on lymph node cells. Natural killer cell activity and the ability to cause local graft-versus-host reaction were not inhibited with CyA treatment. These results suggest that CyA inhibits the proliferation and differentiation of thymocytes, or that CyA makes thymocytes migrate rapidly from cortex to periphery. PMID- 3044649 TI - Humoral immune responses within the lung of bone marrow transplant recipients studied by bronchoalveolar lavage. AB - We investigated 20 bone marrow transplant recipients with pneumonitis using bronchoalveolar lavage (BAL) to assess the humoral immune response in the lung. We measured the levels of total IgG, IgM and IgA in bronchoalveolar lavage fluid and serum, and albumin measurements were used to correct for simple diffusion of immunoglobulins from serum into the lung. We found evidence for the local production of immunoglobulins G, M and A in 15 patients. This was independent of the cause of the pneumonitis. We also found that, although seven patients who recovered from their pulmonary problem had evidence of considerable local production of immunoglobulin, eight patients who died were also producing immunoglobulins in the lung. Death due to pneumonitis in BMT recipients cannot, therefore, be ascribed to a failure of the local humoral immune response. PMID- 3044650 TI - A comparison of the effects of plasma exchange and immunoadsorption on anti insulin antibody synthesis in rabbits. AB - Plasma exchange (PE) and ex vivo immunoadsorption (IA) may be applicable to the removal of anti-insulin antibodies (AI-Ab) from diabetic patients. However, the removal of antibodies may prompt an increase in their rate of synthesis and an overshoot of antibody levels which may be deleterious to the patient. The effects of both PE and IA on AI-Ab synthesis were studied in a rabbit model. Rabbits were immunized with insulin and the resulting AI-Abs removed by both plasma exchange and specific immunoadsorption. Following AI-Ab removal by PE no increase in AI-Ab synthesis or antibody overshoot occurred. However a large increase in AI-Ab synthesis and overshoot occurred following specific AI-Ab removal by immunoadsorption. Despite similar reductions in AI-Ab levels by PE and IA, no increase in antibody synthesis occurred due solely to antibody removal. It is likely that antigen released from the immunoadsorbent stimulated the increase in antibody synthesis following immunoadsorption. These findings are relevant to the clinical application of both PE and IA. PMID- 3044651 TI - Transient increase of serum IgD levels after allogeneic bone-marrow transplantation. AB - Serum IgD levels were followed longitudinally twice a week for up to 100 days in 60 children undergoing allogeneic bone-marrow transplantation (n = 52) or immunosuppression (n = 8) for the treatment of leukaemia, severe aplastic anaemia or severe combined immunodeficiency. In 40 out of the 49 post-transplantation periods analysed (82%), a transient sharp increase of serum IgD was detected, irrespective of initial disease. A similar peak was found in one out of five children after immunosuppressive treatment. A second IgD peak was only recorded in grafted patients (14/49 post-transfusion periods). Peak levels of IgD ranged from 1.3 to 185.7 IU/ml (median 12.2 IU/ml), which represents a 2.6 to 22.4-fold increase over 'baseline' levels. In the transplanted leukaemia and aplastic anaemia patients, the rise of serum IgD occurred at the same time (geometric mean 16 days after transplantation) and was shown to represent heterogeneous polyclonal IgD in six of them. The onset of the serum IgD peak was significantly delayed in children suffering from severe combined immunodeficiency (P less than 0.05) and was demonstrated in one patient to consist of homogeneous IgD. No relation was found between either the occurrence of clinical acute graft-versus host disease or infections after treatment, and the time of onset of IgD elevations. To detect transient serum IgD peaks as described here, frequent sampling of sera is necessary. The origin of the early IgD peaks seems to reside within the recipient's cells by an unknown mechanism. The late IgD peaks are most probably an expression of gradual reconstitution of the immune system following bone-marrow transplantation. PMID- 3044654 TI - [Possibilities of vital bone grafts in the maxillofacial area]. PMID- 3044652 TI - Quantitation of human serum polymeric IgA, IgA1 and IgA2 immunoglobulin by enzyme immunoassay. AB - This report concerns the relative quantitation of serum polymeric IgA and polymeric IgA subclass concentrations by enzyme immunoassay (EIA). The assay relies on the specific binding of polymeric IgA to secretory component. Competition between pentameric IgM and polymeric IgA for binding to secretory component was observed. Thus, samples were adsorbed for IgM by affinity chromatography before the EIA was performed. The assay was used to determine an age-related range of serum polymeric IgA concentrations and to compare the polymeric IgA concentrations in patients with IgA nephropathy (n = 50) to those of controls (n = 50). The serum concentrations of both polymeric IgA and polymeric IgA1 increased with age reaching adult values of around 12 years of age. Polymeric IgA2 concentrations did not reach adult levels until 18 years of age. The ratio of the polymeric IgA concentration to the total serum IgA concentration was found to be significantly increased in children under 2 years of age compared with those over 4 years of age (Mann-Whitney U-test, P less than 0.01). Patients with IgA nephropathy had significantly increased concentrations of polymeric IgA (P = 0.001) and polymeric IgA1 (P = 0.001) but similar polymeric IgA2 concentrations to controls. PMID- 3044653 TI - Antibodies to nuclear lamins in autoimmune liver disease. AB - Antibodies to nuclear lamins were detected in sera of patients with autoimmune liver disease. In indirect immunofluorescence tests, these sera revealed staining of the nuclear periphery. Using isolated nuclei, nuclear matrices, nuclear lamina pore complexes, and chromatographically purified lamins as antigen source, the nuclear lamins A, B, and C were identified as reactive antigens in immunoblotting experiments. The lamins were also identified by 2-D gel electrophoresis. Antibodies to nuclear lamins occurred in 12 of 16 cases of active lupoid hepatitis, but not in 35 patients with the disease in remission. However, only 3 of 37 sera of patients with primary biliary cirrhosis contained anti-lamin antibodies. Autoimmune liver disease sera reacted preferentially with lamins A/C and less frequently with lamin B or lamins A/B/C. PMID- 3044655 TI - [Immunohistochemical studies on palatinal salivary glands]. PMID- 3044656 TI - [Value of imaging procedures in primary tumors of the visceral cranium. 1. Optimizing the diagnostic procedure in surgical treatment planning]. PMID- 3044657 TI - [Tumor-like histiocytic lesions of the oral cavity. A problem in differential diagnosis]. PMID- 3044659 TI - Bilateral simultaneous rupture of the quadriceps tendons. A report of four cases and review of the literature. AB - Four patients with bilateral simultaneous rupture of the quadriceps tendons were each older than 70 years of age. In the first patient the diagnosis was missed and one side was managed conservatively, while the other side was surgically treated. A combination of inadequate postoperative immobilization and the use of chromic catgut sutures probably caused failure of the repair, requiring revision surgery. The other three patients were treated by bilateral operative repair and had satisfactory results. Nonabsorbable suture material use and a minimum period of six weeks of postoperative immobilization are recommended. PMID- 3044658 TI - Generation of adhesive tumor variants: chromosomal changes, reduction in malignancy and increased expression of a distinct membrane glycoprotein. AB - Tumor cell variants which grow adherent to a plastic surface could be isolated in a reproducible way from the high metastatic tumor cell line ESb which grows in a suspension culture. This occurred when starting selection from the uncloned parental line as well as from a freshly derived non-adhesive subclone. The variants showed changes in their karyotype. These were quantitative (tetraploidization) and qualitative (single chromosome aberrations involving the chromosomes 12 and 17 and a marker MX-7). Phenotypic cell surface changes were documented in vitro by immunofluorescence using a monoclonal antibody (mAb 12-15) directed against a distinct plasma membrane glycoprotein of 60-69kD (gp 60-69). The expression of gp 60-69 increased with time of selection for adherence to plastic surface. The adherent cells showed in all cases a greatly reduced overall malignancy as seen by a prolonged survival time of respective tumor bearing animals compared with the suspension growing parental cells. PMID- 3044660 TI - The treatment of forearm fractures with pins. By Georg Schone, 1913. AB - In the case of fractures of the middle one-third of the forearm that are several weeks old, the procedure is as follows. Use a brachial block or general anesthesia. Open the radial fracture by freeing up the fracture site and clearing the medullary canals. About a 4-cm longitudinal incision is made over the dorsal aspect of the radius at the wrist distant from the fracture and between the tendons of the extensor pollicis longus and the extensor carpi radialis longus and brevis. Make a hole in the radius with a small gouge with insertion of the pin to the fracture site and then drive it into the proximal fragment. Then cut the pin off at the right length and set it into the bone with hammer blows to the driver. Remove the driver. Check the reduction of the fracture and closure of the wound. Repeat the procedure on the ulna with the difference that the trephination of the ulna is done proximally. Apply the splint. The fracture site must be protected carefully to avoid angulation. Should this occur, it can be corrected without opening the fracture site because the pins are flexible. We have operated upon four isolated fractures of the ulna, one isolated fracture of the radius, and two fractures of both radius and ulna. I believe that the procedure is simple and practical to perform. Whether and to what extent a similar procedure can be carried out satisfactorily on other bones with larger marrow canals remains to be seen. PMID- 3044661 TI - Sonographic evaluation of the rotator cuff. Accuracy in patients without prior surgery. AB - Since 1982 a sonographic technique has been devised to evaluate the integrity of the rotator cuff. In a series of 139 shoulders in 134 consecutive patients without prior surgery, sonographic results were correlated with surgical findings and/or arthrography. When sonography was compared with surgical findings (n = 90 shoulders), the overall accuracy was 95%. The sonographic diagnosis was the same as the arthrographic diagnosis in 91% of the cases (n = 50). Cuff lesions were also created in two cadavers, and images were made to confirm sonographic appearances. Because in the authors' experience sonography is accurate, safe, painless, rapid, and inexpensive, it has now been adopted as the primary study for the examination of the rotator cuff at the authors' institution. Arthrography is recommended in cases in which sonography is negative but there is significant clinical evidence of rotator cuff tear. PMID- 3044662 TI - Acute compartment syndrome in the thigh. A case report and a review of the literature. AB - Compartment syndrome in the thigh is an uncommon condition, and acute compartment syndrome without an associated fracture is even less common. An 18-year-old male developed an acute anterior compartment syndrome in the thigh from a contusion without an associated fracture. This syndrome was treated successfully with a fasciotomy. PMID- 3044664 TI - Management of open fractures with sterilization of large contaminated, extruded cortical fragments. PMID- 3044665 TI - Atrial systole: its role in normal and diseased hearts. PMID- 3044663 TI - Ectopic bone formation is enhanced in senescent animals implanted with embryonic cells. AB - Ectopic bone formation induced by the subcutaneous implantation of demineralized bone matrix (DBM) is very significantly reduced in older Fischer 344 rats. Cells originating from calvaria of 20-day-old embryo donors were introduced into cylinders of DBM sealed at the ends with a Millipore filter or collagen sponges prior to subcutaneous implantation. Cells within the chambers had access to vascular channels that could penetrate through the interstices of the DBM. After four weeks of implantation in 26-month-old rats, the cylinders were full of bone. This bone was assessed by histologic techniques, by calcium and bone gamma carboxyglutamic acid (gla) protein (BGP) concentrations, and by alkaline phosphatase activity. Cylinders to which no cells were added produced no bone. Bone marrow cells enclosed in similar cylinders or injected weekly at the implantation site also enhanced new bone formation but to a much lesser extent. Embryonic muscle cells formed large amounts of cartilage and less bone. Fibroblasts were inactive in this system. Prior treatment of the DBM with trypsin inhibited the myoblast response but not that of calvaria cells. PMID- 3044666 TI - Effects of hyperinsulinaemia on the cardiovascular responses to graded hypovolaemia in normal and diabetic subjects. AB - 1. Two experiments were carried out. The first with five normal male subjects was placebo controlled and single blind, each subject being studied on two occasions. Lower body subatmospheric pressure (LBSP) was used to assess the cardiovascular effects of graded hypovolaemia before and during either a hyperinsulinaemic, euglycaemic clamp or a placebo clamp using 0.9% (w/v) NaCl only. 2. During hyperinsulinaemia, resting systolic blood pressure rose and was accompanied by forearm vasodilatation. Forearm blood flow (FABF) and heart rate (HR) were higher at each level of LBSP during than before hyperinsulinaemia. In addition, hyperinsulinaemia was accompanied by a small increase in noradrenaline, but packed cell volume did not change. 3. In the second experiment, the effects of a hyperinsulinaemic euglycaemic clamp on the cardiovascular responses to LBSP were assessed in seven diabetic subjects with peripheral and autonomic neuropathy. 4. In contrast to the normal subjects, there was a slight fall in systolic blood pressure during the clamp but no effect was noted on HR or FABF. Mean arterial blood pressure was lower at each level of LBSP during hyperinsulinaemia compared with the pre-clamp period. Packed cell volume fell during the clamp and plasma noradrenaline rose. In one of the diabetic subjects, a precipitous fall in blood pressure occurred during hyperinsulinaemia when LBSP of 10 mmHg (1.3 kPa) was applied, this manoeuvre having been well tolerated before the clamp. 5. The mode of action of hyperinsulinaemia is not clear, but there was, however, no evidence that a fall in plasma volume had occurred. PMID- 3044667 TI - Secondary ciliary dysfunction. PMID- 3044668 TI - Differences in glomerular binding and response to angiotensin II between normotensive and spontaneously hypertensive rats. AB - 1. Angiotensin II (ANG II) binding and the physiological response to exogenous ANG II have been studied in isolated glomerular preparations from normotensive (NTR) and spontaneously hypertensive (SHR) rats. 2. The binding of 125I-labelled ANG II by glomeruli from SHR was significantly greater than that by glomeruli from NTR, whereas the binding affinity constant (Ka) showed that the SHR ANG II glomerular receptor had a lower affinity for the hormone than the NTR glomerular receptor. 3. Glomeruli from SHR were significantly less responsive to exogenous ANG II than those from NTR. 4. Sodium loading resulted in a significant increase in ANG II binding by glomeruli from NTR, whereas a decrease in binding occurred in glomeruli from SHR. 5. Although a high sodium intake caused a reduction in the response of glomeruli from both NTR and SHR to exogenous ANG II, these changes were not statistically significant. In NTR this was associated with a decrease in the concentration of agonist required to cause half-maximal response (EC50), whereas an increase in EC50 was shown by glomeruli from SHR. PMID- 3044669 TI - Relief of spasm with diclofenac. PMID- 3044671 TI - Diagnostic significance of salivary levels of beta 2-microglobulin in Sjogren's syndrome. AB - To evaluate the diagnostic significance of salivary beta 2m in Sjogren's syndrome we measured salivary beta 2m levels in 19 patients with primary sicca syndrome (PSS), 15 with secondary Sjogren's syndrome (SSS) and compared the results with 20 normal healthy persons. We showed that beta 2m is specifically excreted in the saliva, because in normal saliva the concentration of beta 2m was unrelated to IgA levels. Also in normals, there was no relationship between serum and saliva concentrations of beta 2m. The mean saliva levels of beta 2m were increased in PSS (1.13 +/- 0.58) and SSS (1.39 +/- 0.69) compared with the levels in normals (0.53 +/- 0.22). The determination of beta 2m in the saliva can therefore be used as a noninvasive measurement for the confirmation of the diagnosis Sjogren's syndrome. PMID- 3044670 TI - Bone metabolism in patients with rheumatoid arthritis. AB - Bone metabolism in patients with rheumatoid arthritis is reviewed. Two different entities are recognised: 1) a localised periarticular bone loss, due to inflammatory processes and 2) a generalised increased bone turnover, ultimately leading to a loss of axial bone mass. The mechanism of this loss of axial bone is not completely understood; probably immobilisation is the most important factor. The influence of certain drugs is discussed. PMID- 3044672 TI - Dialysis-related beta 2 microglobulin-amyloid arthropathy. Improvement of clinical symptoms after a switch of dialysis membranes. AB - Amyloidosis containing beta 2 microglobulin (beta 2m) and joint pains are frequent complications of long-term hemodialysis. In a prospective study the authors planned to act on dialysis arthropathies (DA) by improving beta 2m depuration: for 8 patients suffering from DA we replaced usual cuprophane membranes (CU) by high permeability polyacrylonitrile membranes (PAN). DA was diagnosed on the presence of severe disabling arthralgias in the absence of any other obvious etiology, and on the presence of at least 3 among 6 criteria. The efficacy of the switch of DM was assessed on 6 criteria including the patient's own evaluation at the end of the study and a joint pain severity index. Clinical evaluation and beta 2m measurement were carried out at the start of the trial and after a mean duration of dialysis on PAN of 6 months. An improvement in joint symptomatology and a decrease of joint severity index were noted in the 8 cases. Our results suggest that the change of DM has a favorable influence on DA. This improvement could result either from a better beta 2m depuration or from a better biocompatibility of PAN than the CU previously used. PMID- 3044673 TI - A double blind randomised placebo controlled trial of hexopal in primary Raynaud's disease. AB - The peripheral vasospastic symptoms associated with Raynaud's disease continue to be an unsolved clinical problem. Hexopal (Hexanicotinate inositol) has shown promise in uncontrolled studies and its use in patients with Raynaud's disease may reduce such vasospasm. This study examines the effects of 4 g/day of Hexopal or placebo, during cold weather, in 23 patients with primary Raynaud's disease. The Hexopal group felt subjectively better and had demonstrably shorter and fewer attacks of vasospasm during the trial period. Serum biochemistry and rheology was not significantly different between the two groups. Although the mechanism of action remains unclear Hexopal is safe and is effective in reducing the vasospasm of primary Raynaud's disease during the winter months. PMID- 3044674 TI - Ventilation-perfusion inequality during endotoxin-induced pulmonary vasoconstriction in conscious sheep: mechanisms of hypoxia. AB - In sheep low-dose endotoxin infusion causes transient thromboxane-mediated pulmonary vaso- and bronchoconstriction. These changes are paralleled by a decrease of arterial PO2. To elucidate the mechanisms of hypoxia we analysed ventilation-perfusion (VA/Q) distributions by the multiple inert gas elimination technique before and after endotoxin infusion in six conscious sheep. We found that hypoxia after endotoxin could be explained by both a VA/Q inequality and a simultaneous decrease in the mixed venous oxygen tension. The latter was due to a decrease of cardiac output. True shunt did not contribute to hypoxia as it was unchanged. PMID- 3044675 TI - Glomerular filtration rate in adults estimated from 123iodine-hippuran and 99mtechnetium-diethylenetriaminepenta-acetic acid gamma camera renography. AB - In a retrospective study a close relationship was found between the rate constant for renal clearance of the radioactive indicator (lambda pk) and the glomerular filtration rate (GFR) measured by 51Cr-EDTA plasma clearance. The material comprised eighteen adult subjects submitted to 123I-hippuran gamma camera renography (IHGR) and twenty-two adult subjects to 99mTc-DTPA gamma camera renography (TDGR). The rate constant was calculated from a bi-exponential decomposition of the activity-time curve recorded within a small region of interest over the left ventricle. The total cleared renal fraction (TCRF) of the cardiac output with respect to the radioactive indicator has previously been shown to be closely related to GFR. A pooled estimate of GFR (GFRp) was calculated from the stochastically independent estimates of GFR based on lambda pk and TCRF. The comparison of GFRp with measured GFR was satisfactory and yielded substantially smaller standard deviations (SD) of GFRp than estimates based on lambda pk and TCRF separately. The standard deviations of GFRp were about 7 and 12 ml/min/1.73 m2 in IHGR for GFR equal to 50 and 100 ml/min/1.73 m2, respectively. The corresponding SD in TDGR were about 7 and 11 ml/min/1.73 m2. These standard deviations are sufficiently small for many clinical purposes and the method requires no blood samples or urine collections. PMID- 3044676 TI - Plasma renin activity and oxygen content along the renal veins in hypertensive patients. AB - Plasma renin activity (PRA) and oxygen content in three different locations in the renal veins were studied in 27 patients with suspected renin-dependent hypertension or during post-operative follow-up investigations after renal artery reconstruction. A significant decrease in PRA was found along the right renal vein but not along the left one. The oxygen content simultaneously measured was lower in the central parts of both renal veins than most peripherally, more on the right side than on the left one, indicating dilution of renal venous blood. An erroneous ratio between renal vein PRA of the affected and the contralateral kidney was found in two of 17 patients with unilateral renal artery stenosis when sampling from a central position in the renal vein. This indicates that a peripheral position of the catheter is important when sampling for renal vein PRA. PMID- 3044678 TI - Pneumocystis carinii. AB - This article covers the biology, epidemiology, and pathology of infection with Pneumocystis carinii. The clinical presentation, course, and treatment of pneumonia caused by this organism in patients infected with the human immunodeficiency virus is discussed. PMID- 3044677 TI - Pulmonary immunology. AB - The lower respiratory tract of normal healthy individuals is kept sterile and disease-free by formidable local and systemic immune system defenses. In patients with AIDS, these defenses are severely compromised. Undoubtedly, alterations in both local and systemic forces make the host vulnerable to infection by opportunistic organisms. Indeed, substantial evidence suggests that at least a small percentage of macrophages, which normally function alone and in concert with other immune components, can be and are infected with HIV, and do not function normally. Abnormalities with lymphocytes in the lung may contribute to macrophage defective function. Further investigations into the role of lung macrophages and other lower respiratory tract defenses are needed if we are to comprehend fully the mechanisms that lie behind the multitude of pulmonary complications associated with AIDS. PMID- 3044679 TI - Mycobacterial disease: epidemiology, diagnosis, treatment, and prevention. AB - TB is common in the setting of HIV-induced immunosuppression, especially among demographic groups with a high background prevalence of tuberculous infection. It is often the first (sentinel) infectious disease to appear, extrapulmonary and disseminated disease is common, the chest x-ray picture is frequently atypical, and the tuberculin skin test is often falsely negative. It therefore requires a high index of suspicion and an aggressive diagnostic approach to avoid missing HIV-related tuberculous disease, which is communicable from man to man by the aerosol route and which appears to be highly treatable with conventional anti-TB drugs. Identification and INH prophylaxis of tuberculous-infected, HIV seropositive persons is likely to be very important in the prevention of tuberculous disease. MAI is also a very common pathogen that frequently produces extrapulmonary and disseminated disease among patients with AIDS. In contrast to TB, AIDS-related MAI disease occurs more uniformly among the AIDS risk groups, occurs late among the HIV-related infections, and is not effectively treated with current drug regimens. PMID- 3044680 TI - Bacterial and fungal pneumonias. AB - Bacterial pneumonias occur with increased frequency and can be associated with increased morbidity in the HIV-infected population compared with normals. The pathogens that most frequently cause community-acquired pneumonias are S. pneumoniae, H. influenzae, and occasionally S. aureus. These pneumonias usually respond to appropriate antibiotic therapy; however, patients diagnosed with bacterial pneumonias are at increased risk for subsequent episodes. Nosocomial pneumonias, by contrast, are usually caused by gram-negative organisms and have a high mortality. Fungal pneumonias also have an increased incidence in AIDS patients, and usually occur in the setting of disseminated disease. Infections caused by C. neoformans, H. capsulatum, and C. immitis often recur despite a good initial response to amphotericin B. Maintenance therapy with an antifungal agent is therefore recommended. PMID- 3044681 TI - Kaposi's sarcoma. AB - Pulmonary KS may occur in up to 20 to 25 per cent of patients with cutaneous KS. The presenting symptoms of pulmonary KS are indistinguishable from those of opportunistic pathogens that cause pneumonia. It most frequently presents with the symptoms of cough or dyspnea; however, fever, hemoptysis, and stridor have been reported to occur secondary to pulmonary KS. Roentgenographically, pulmonary KS may present as diffuse infiltrates, nodal disease, and/or pleural effusions. The diagnosis is established by a characteristic histologic pattern obtained from large pieces of tissue, that is, from open lung biopsy or autopsy rather than from transbronchial biopsy. No effective therapy for pulmonary KS exists; however, short-term palliation may be achieved with radiotherapy or combination chemotherapy. In a patient with known pulmonary KS who develops either new or changing symptoms or new roentgenographic findings, an attempt should be made to rule out an associated infectious process. PMID- 3044682 TI - Lymphocytic interstitial pneumonia. AB - Lymphocytic interstitial pneumonia is a common complication of HIV infection in children, but uncommon in adults. It is characterized clinically by the presence of cough and dyspnea, diffuse pulmonary infiltrates on chest x-ray, restrictive pulmonary dysfunction, and hypoxemia. This constellation of findings usually erroneously suggests PCP, and a lung biopsy is necessary to establish the diagnosis. Typical microscopic findings include diffuse infiltration of the pulmonary interstitium with a mixture of lymphocytes and plasma cells; immunohistologic studies reveal that in association with HIV infection, these lymphocytes are T cells. The pathogenesis of LIP in patients with HIV infection is not known. It is believed that it represents a tissue response to EBV infection, HIV infection of the lung, or both. Although patients with LIP may respond dramatically to corticosteroid therapy, others may improve with no treatment. Unfortunately, most patients eventually succumb to other complications of HIV infection. PMID- 3044683 TI - Pulmonary function tests. AB - The clinical significance of pulmonary function tests (including blood gas analysis) lies in their sensitivity for detecting PCP. PCP has most consistently been found to cause abnormalities in the DLCO and the exercise arterial blood gas; both are highly sensitive for the presence of Pneumocystis infection. These tests are more sensitive for the detection of PCP than are the resting arterial blood gas and chest x-ray. Therefore, measuring these values can be especially helpful in evaluating HIV-infected individuals who have pulmonary symptoms but whose resting arterial blood gas and/or chest radiograph are normal. The advantage of performing the exercise test over measuring the DLCO is that the exercise test is simple. It can be done without pulmonary function equipment and without a technologist. Furthermore, since many AIDS patients with non-PCP pulmonary disorders maintain "normal" exercise tests despite abnormal DLCO, it can be useful in evaluating patients for PCP who have known underlying lung disease with progressive symptoms. Measurement of lung volumes and spirometry lacks both sensitivity and specificity for detecting pulmonary disease in general and PCP in particular. Spirometry is helpful in detecting airways obstruction, which is not an uncommon finding in AIDS patients. The etiology, clinical significance, and treatment of obstructive ventilatory defects in the AIDS population remains unclear. PMID- 3044684 TI - The radiology of pulmonary disease. Chest radiography, computed tomography, and gallium scanning. AB - A review of the radiologic manifestations of AIDS pulmonary diseases, with an emphasis on the utility of gallium scanning in the context of the normal or equivocal chest x-ray, is presented. PMID- 3044685 TI - Diagnosis of pulmonary diseases. AB - A broad spectrum of lung disease occurs in association with HIV infection. Included are both infectious and neoplastic processes and idiopathic disorders. To insure prompt, accurate, and efficient diagnosis, a logical, staged sequence of tests should be applied. Chest films and, in some instances, pulmonary function tests and gallium-67 citrate lung scans serve to provide objective indications of lung disease. Each of these tests is sensitive but nonspecific. Specific infecting organisms, particularly P. carinii, can be identified by examining sputum induced by inhalation of 3 per cent saline. Bronchoscopic procedures, including BAL and TBB, are highly sensitive and should be performed in patients having nondiagnostic sputum examinations. Tests involving antigen and antibody detection are of little use in the evaluation of individual patients. Detection of recurrent episodes of PCP is difficult because abnormalities in the usual screening tests may be residual from previous episodes. Finding P. carinii in sputum or bronchoscopic specimens soon (within 2 to 3 months) after a confirmed episode of PCP likely represents residual organisms rather than recrudescence of the infection. Empiric diagnosis of P. carinii should be employed only in limited circumstances when specific diagnostic studies are not available, are contraindicated, or are refused. PMID- 3044686 TI - Pulmonary disease in infants and children. AB - Pulmonary diseases contribute significantly to the morbidity and mortality in children infected with the human immunodeficiency virus. The wide array of lung diseases spans from early acute bacterial infections, to the lymphoid interstitial pneumonitis/pulmonary lymphoid hyperplasia complex, to opportunistic infections. The unique clinical and histopathologic features of these diseases are reviewed. PMID- 3044687 TI - Pulmonary effects of AIDS: nosocomial transmission. AB - This review provides an overview of the risk of nosocomial infection in the "AIDS era." Airborne spread of Mycobacterium tuberculosis from affected patients has re emerged as a hazard to hospital personnel. The risk of acquiring clinical illness due to Pneumocystis carinii or cytomegalovirus is, in contrast, a function of the immunocompetence of the health care worker. Methods of transmission as well as the epidemiology of human immunodeficiency virus-related infection in the health care worker will be discussed. The increase in the number of immunocompromised patients (AIDS and non-AIDS) requires careful attention to infection control methodology with respect to the cleansing of the fiberoptic bronchoscope, the intensive care unit's respiratory equipment (such as mechanical ventilators and nebulizers), and the pulmonary function laboratory. PMID- 3044688 TI - Oral hygiene behavior and periodontal status in European adolescents: an overview. AB - The object of this review is to examine toothbrushing as a health behavior in European adolescents. The aims are to investigate the role of environmental and social factors on toothbrushing as a health behavior, to illustrate the importance of such factors in the prevention of periodontal diseases and to demonstrate how a knowledge of these factors can aid the development of specific health education programs. In order to do this the results of many studies concerned with oral hygiene behavior from different European countries have been examined. It is from the detailed study of these various European investigations that the results section is composed. The findings from this review demonstrate that toothbrushing as a health behavior is influenced by both environmental and social factors, that both of these factors affect the incidence of gingivitis and periodontal disease in the countries examined, and that when they are incorporated into health education programs based on the community development approach they can aid behavior change. PMID- 3044689 TI - Biological 1H NMR spectroscopy. AB - Proton nuclear magnetic resonance spectroscopy (1H NMR) is a powerful analytical method used to identify and quantitate chemical compounds. In recent years, it has been used to study rates of metabolism in microbes, isolated perfused tissues, intact animals, and human beings. This review highlights some of the more recent biological applications of 1H NMR in the study of metabolic pathophysiology in animals and man. 1H NMR can rapidly analyze complex mixtures of metabolites found in body fluid and biopsy specimens. In vivo 1H NMR methods can measure intracellular pH, a wide variety of metabolites, tissue perfusion, and rates of metabolism of endogenous and exogenous compounds. Using 13C labeled compounds or magnetization transfer techniques metabolic fluxes may be measured in vivo during virtually all normal and abnormal physiological conditions. PMID- 3044690 TI - Comparative studies of glutathione reductase and lipoamide dehydrogenase. AB - 1. Glutathione reductase and lipoamide dehydrogenase are structurally and mechanistically related flavoenzymes catalyzing various one and two electron transfer reactions between NAD(P)H and substrates with different structures. 2. The two enzymes differ in their coenzyme and functional specificities. Lipoamide dehydrogenase shows higher coenzyme preference while glutathione reductase displays greater functional specificity. 3. Binding preference of the two flavoenzymes for nicotinamide coenzymes is demonstrated by 31P-NMR spectroscopy. 4. The presence of arginines in glutathione reductase which is inactivated by phenyl glyoxal, is likely to be responsible for the NADPH-activity of glutathione reductase. 5. The substrate binding sites of the two enzymes are similar, though their functional details differ. 6. The active-site histidine of glutathione reductase functions primarily as the proton donor during catalysis. While the active-site histidine of lipoamide dehydrogenase stabilizes the thiolate anion intermediate and relays a proton in the catalytic process. PMID- 3044691 TI - Methodological and strategical considerations in schizophrenia research. PMID- 3044693 TI - 40 years of the NHS. Child care: parents rule, OK. PMID- 3044692 TI - Differential diagnosis of anorexia nervosa and depressive illness: a review of 11 studies. AB - A review of differential diagnostic information from 11 studies of patients with anorexia nervosa is provided. Both intercurrent depressive and obsessive compulsive features are most frequently reported overall. In seven of these studies providing information about premorbid and intercurrent personality disturbances, obsessive-compulsive characteristics are reported as most frequent in four. It is suggested that presumed connections between anorexia nervosa and depressive illness may be secondary to a more direct link with the obsessive compulsive syndrome. PMID- 3044694 TI - Labeling guidance text for combination oral contraceptives. Physician labeling. PMID- 3044695 TI - Sonographic signs of early pregnancy. AB - The early embryo (blastocyst) is implanted about 6 to 7 days after fertilization and becomes completely imbedded within the decidua at 9.5 days. As the exocoelomic cavity (or early gestational sac) enlarges, it becomes visible by ultrasonography. The gestational sac, however, remains within markedly thickened ipsilateral decidua. The uterine cavity remains relatively undistorted (i.e., straight) and can be delineated by ultrasonography. The gestational sac, therefore, can be seen adjacent to a straight uterine cavity within a thickened decidua. This is called an intradecidual sign. Before the appearance of a visible gestational sac, an echogenic area within the thickened decidua may be seen. This is the earliest sign of intrauterine gestation, and it can be seen as early as 25 days menstrual age. The earliest evidence of an embryo is seen about 2 weeks later, when two small bubbles (amniotic and yolk sacs) attach to the wall of the gestational sac. The embryonic disc is located between the two bubbles. This is called the double-bleb sign. The intradecidual and double-bleb signs represent the most important signs of early intrauterine pregnancy. PMID- 3044697 TI - Biomedical versus cultural constructions of abnormality: the case of idiopathic hirsutism in the United States. AB - From an examination of (1) psychological and social stresses documented by the medical profession of women with amounts of hair departing from what is considered to be the "feminine" distribution of hair (idiopathic hirsutism), (2) biomedical information about male and female hair growth and distribution, (3) definitional influences of society, and (4) instrumental maneuverings of depilatory industries, this paper offers insights about the alteration of natural female facial and body hair to conform to a social construct of gender status. Although the hair problem is interesting in itself, its implications are much broader, for what lies behind conceptions of abnormality is a biological reality mediated by social constructs and classificatory schema. PMID- 3044696 TI - Cryopreservation and transplantation of hepatocytes: an approach for culture and clinical application. AB - The development of a reproducible, effective system of cryopreservation of hepatocytes would create new possibilities for metabolic studies, as well as the clinical application of hepatocellular transplants in diverse hepatic disorders. For the purpose of dealing with both questions, we have studied different media and procedures of freezing, and have chosen that combination which afforded the best results for long-term studies of function. A series of intrasplenic transplants were performed with freshly isolated hepatocytes as well as cryopreserved ones, and several hepatospecific parameters (Alanine aminotransferase, ornithine carbamoyl transferase, lipogenesis, uptake of 99mTc-N (p-butylphenylcarbamylmethyl)iminodiacetic acid, G6Pase, albumin, and gamma glutamyl transpeptidase (gamma-GT) were quantified for 9 months. The histological study of the cells reveals that they acquired an architecture characteristic of the liver. All of the parameters indicative of hepatic function were detected throughout the 9-month period in the transplanted spleens. All the spleens, controls and transplanted, were found negative for the presence of the dedifferentiation marker gamma-GT. These results clearly indicate the possibility of recovering hepatocytes which maintain their specific functions after being subjected to a freezing-thawing process, with the corresponding implications for future clinical application as well as for hepatic biochemistry. PMID- 3044698 TI - Accurate localization of ruptured sinus of Valsalva aneurysm by real-time two dimensional Doppler flow imaging. AB - The surgical approach to the repair of a ruptured sinus of Valsalva aneurysm can depend on the cardiac chamber into which rupture occurs. This report details the color flow Doppler images in two patients who developed a right sinus of Valsalva aneurysm to right atrial fistula owing to bacterial endocarditis. In both cases, the color flow Doppler image was superior to contrast aortography in identifying the chamber into which rupture had occurred. The early experience with real-time two-dimensional Doppler flow imaging suggests that this noninvasive technique is valuable in the management of ruptured sinus of Valsalva aneurysms. PMID- 3044699 TI - Provocation dose and discriminant analysis in histamine bronchoprovocation. Are the current predictive data satisfactory? AB - In 20 normal subjects (N) and 20 asthmatic patients (A) using bronchodilators as needed, the PD40, PD10, PD15, PD20, PD10T, and PD20C were measured (PD being provocation dose, subscript being -% delta in Gaw/VL for PD40, lowest FEV1 for PD20C, and best FEV1 for the remaining PDs; 10T means delta FEV1 greater than -10 percent). For discriminant analysis we used an ad hoc graphic (best case) method, a ceiling method based on highest PD in A and two methods (logistic and linear) which considered PDs in both N and A (PDN and PDA, respectively). The distribution of PDN and PDA had substantial overlap and appeared log normal. The PD15, PD20, and PD20C displayed the smallest mean misclassification error followed by PD10T, PD40, and PD10. The linear and logistic methods produced balanced sensitivity and specificity but, predictably, a misclassification error higher than that of the graphic method. The ceiling method proved unsatisfactory with 100 percent sensitivity but approximately equal to 60 percent specificity. Using linear and logistic methods, the posttest likelihood of asthma could be expressed as a function of its pretest likelihood and level of PD recorded. We concluded that: (1) the most discriminant PDs are PD20, PD15 and PD20c; and (2) new normative data for diagnostic bronchoprovocation are needed, because: (a) when PDA and PDN overlap, the currently used ceiling method leads to a high misclassification rate, while the linear and logistic method based on mathematical model have a better discriminant ability; (b) to separate PDA from PDN and allow the application of the ceiling method, "as-needed bronchodilators" is not a reproducible criterion. PMID- 3044700 TI - Significance of T-wave pseudonormalization during exercise. A radionuclide angiographic study. AB - In 84 consecutive patients with resting T-wave inversion, radionuclide angiography revealed significant new wall motion abnormalities in 13 (28 percent) of the 47 patients with persistent T-wave inversion and in 23 (62 percent) of the 37 patients with T-wave pseudonormalization during exercise (p less than 0.01). The response of the ejection fraction to exercise was better in patients with persistent T-wave inversion than in those with pseudonormalization (p less than 0.04). Mechanical evidence of ischemia was seen in 14 (61 percent) of the 23 patients with T-wave pseudonormalization but without ST-segment depression. In patients with resting T-wave inversion, pseudonormalization was slightly more sensitive but less specific than a positive exercise test for predicting significant new wall motion abnormalities or decreases in the ejection fraction with exercise. Although pseudonormalization is not extremely useful alone, the presence or absence of this finding can increase the diagnostic accuracy of exercise electrocardiography in patients with resting T-wave inversion and suspected ischemic heart disease. PMID- 3044701 TI - Neutrophil chemotactic factor release and neutrophil alveolitis in asbestos exposed individuals. AB - Alveolar neutrophil accumulation occurs in asbestosis. To evaluate a possible role for release of neutrophil chemotactic factor (NCF) in the pathogenesis of asbestosis, spontaneous NCF release from alveolar macrophages obtained by bronchoalveolar lavage (BAL) in eight individuals with asbestosis, 13 asbestos exposed individuals without asbestosis, and five control subjects has been studied. Alveolar macrophages were incubated in medium (four hours; 37 degrees C), and neutrophil responses to the supernatants were assayed in a microchemotaxis chamber. Alveolar macrophages from subjects with asbestosis released more NCF (97 +/- 19 neutrophils per high-power field [N/HPF]) than controls (3 +/- 1 N/HPF; p less than 0.01). Alveolar macrophages from individuals with asbestos exposure and increased BAL neutrophil proportions (n = 7) released more NCF (93 +/- 24 N/HPF) than individuals with asbestos exposure and normal BAL neutrophil proportions (n = 6; 11 +/- 6 N/HPF; p less than 0.02). The results show that spontaneous NCF release occurs in asbestosis and that NCF release is associated with neutrophil alveolitis in asbestos-exposed individuals without asbestosis, suggesting a pathogenic role for NCF in mediating this neutrophil alveolitis. The results of the study also suggest that the presence of crackles is a better predictor of the presence of neutrophil alveolitis than is an abnormal chest x-ray film. PMID- 3044702 TI - Influence of PEEP ventilation immediately after cardiopulmonary bypass on right ventricular function. AB - Ventilation with positive end-expiratory pressure (PEEP) is often the appropriate therapy for treating patients with impaired pulmonary function after cardiac surgery procedures. Circulatory depression, however, sometimes limits the level of PEEP. This study was conducted to investigate the effects of PEEP ventilation (+15 cmH2O) immediately after weaning from cardiopulmonary bypass 1) period of PEEP application and 45 min thereafter; 2) period of PEEP application on right ventricular hemodynamics using a new thermodilution technique for measuring right ventricular ejection fraction (RVEF), right ventricular end-diastolic and end systolic volumes (RVEDV, RVESV). Forty patients undergoing aortocoronary bypass grafting were retrospectively divided into two groups: group 1 (n = 24) in which RVEF was reduced significantly (40----28 percent), and group 2 (n = 16) in which RVEF remained almost unchanged. In patients in group 1, stenosis of the right coronary artery (RCA) was significantly more pronounced in comparison to the others and was detected to be responsible for the different reaction of RVEF (analysis of co-variance). Application of PEEP immediately after weaning from CPB was followed by an increase in RVESV (+4 percent; RVEDV -1 percent) in group 1, whereas patients of group 2 differed significantly (RVESV -14 percent; RVEDV -15 percent). Cardiac index was decreased only in group 1 (-32 percent). During the second period of PEEP application, no further difference could be observed between the groups. We conclude that hemodynamic changes related to PEEP ventilation are minimal in the intact right ventricle. Abnormalities in right ventricular function due to stenosis of the RCA, however, have had marked clinical influence on the circulatory response. Monitoring of right ventricular function seems to be of benefit for cardiac surgery patients in this situation. PMID- 3044703 TI - Lobar collapse. Usual and unusual forms. PMID- 3044704 TI - [Use of the skin-fascia-tendon island flap from the forearm in one-stage reconstruction of tissues of the dorsum of the hand. Case report]. PMID- 3044706 TI - Potential involvement of retroviral elements in human dementias. AB - Creutzfeldt-Jakob disease (CJD) is a dementia of humans caused by a class of infectious agents with several biological properties similar to those of conventional viruses. The molecular nature of this group of agents is enigmatic, for neither an agent-specific nucleic acid nor a non-host protein has yet been identified. Recent transmissions of familial CJD dementias to rodents suggest that this class of agent can be integrated into the germline. Furthermore, tissue culture studies indicate that CJD causes transformation of cells in a manner reminiscent of slowly oncogenic retroviruses. Currently characterized retroviral like elements include many forms that do not have 'typical' retroviral ultrastructural morphology; several forms are also known to be resistant to various types of standard physicochemical inactivation. We suggest that CJD agents are either constituted by retroviral-like nucleic acids or interact with endogenous retroviral sequences to elicit a slowly progressive disease of the central nervous system. Several overlapping properties between infectious CJD and 'non-infectious' dementias, such as Alzheimer's disease, implicate potential common pathogenic mechanisms. PMID- 3044707 TI - Neuropathology of unconventional virus infections: molecular pathology of spongiform change and amyloid plaque deposition. AB - To the triad of neuronal loss, gliosis and spongiform change as characteristic morphological changes associated with infection of the central nervous system, one can now add the presence of scrapie-associated filaments (SAF)/PrP rods. While the host's immune response is conspicuous by its absence, the vigorous astrocytic response is presumptive evidence of the host's ability to recognize and respond to the primary neuronal insult. We assume that the spongiform change and vacuolation of neurons are of fundamental importance in the pathogenesis of the disease, realizing that neither is specific or essential for the replication of the infectious agent. The topographical distribution of lesions is partly explained by the portal of entry and retrograde spread of the virus. The temporal progression of the lesions is more clearly determined by the host genes, best illustrated by studies of the incubation period. The molecular basis of the spongiform change is unknown but it is presumed to involve some disturbance of membrane metabolism. The recognition of PrP as a membrane glycoprotein invites proposals for its role in the development of these spongiform lesions. Extracellular amyloid occurs as plaques or congophilic angiopathy in some instances, and provides the best evidence that Alzheimer's disease (AD) is in some way related to the unconventional virus diseases. However, the protein subunit (A4) of the amyloid fibril in AD and its precursor are quite distinct from the PrP subunit which constitutes the amyloid fibril in these infectious diseases. It is still unclear whether the PrP subunit in the SAF has exactly the same composition as in the extracellular amyloid fibril. Our results suggest that only a fragment of the PrP molecule is the major constituent of the extracellular fibril. Since both PrP and A4 are derived from membrane glycoproteins, the elucidation of their normal function is likely to lead to a better understanding of the spongiform and amyloidogenic lesions in these diseases. PMID- 3044705 TI - Detection of distinct structural domains within the primary constriction using autoantibodies. AB - We report the immunological differentiation of structures within the primary constriction. These include the kinetochore and the connecting strand, a structure which connects sister kinetochores. The location and temporal appearance of the connecting strand antigen suggest that it could play a role in the maintenance of sister chromatid pairing. In addition, we report the identification of a novel epitope that is localized to discrete patches along the entire length of the junction between sister chromatids at metaphase (the junction patch antigen). The patches on the inner surface of the euchromatic arms can be disrupted by Colcemid treatment while those found in the primary constriction remain intact. The apparent heterogeneity of the patches suggests that they may play different roles in the regulation of sister chromatid pairing. Because of their cytological localization and possible functional role, the junction patch and connecting strand antigens have provisionally been collectively termed CLiPs (Chromatid Linking Proteins). All of these antigenic sites are shown to be distinct from centromeric heterochromatin, which can itself be immunologically differentiated from the euchromatic arms. The relationship between the antigenicity of the primary constriction and the unique manner in which chromatin is organized in this region is discussed. PMID- 3044709 TI - Genetic aspects of unconventional virus infections: the basis of the virino hypothesis. AB - The properties of genes involved directly or indirectly in the pathogenesis of scrapie and other unconventional (UCV) virus infections are reviewed. Reasons are presented for assigning paramount importance to the Sinc gene in mice and the Sip gene in sheep (the likely homologue of Sinc). The rationale is given for concluding that the agents of UCV infections have their own genomic molecules coding for strain differences. The virino hypothesis, which proposes that the infective form of the agent is an informational hybrid between the agent's genome and protective host proteins, is presented in detail, with an explanation of the postulated role of Sinc. PMID- 3044710 TI - Abstracts for the XIII international meeting of the Society for Analytical Cytology. September 4-9, 1988, Breckenridge, Colorado. PMID- 3044708 TI - The clinical neurology and epidemiology of Creutzfeldt-Jakob disease, with special reference to iatrogenic cases. AB - The clinical characteristics of Creutzfeldt-Jakob disease (CJD) in a newly analysed group of 223 cases transmitted to primates at the NIH are compared to a recent large series of neuropathologically verified cases in France, and the limited conclusions from worldwide epidemiological studies are briefly summarized. Discussion then focuses on iatrogenic CJD, with special attention to the interplay of clinical, laboratory and epidemiological features of the current outbreak of CJD in hypopituitary dwarfs treated with growth hormone extracted from pools of human pituitary glands. PMID- 3044711 TI - Nonselective enrichment for yeast adenine mutants by flow cytometry. AB - The expression of certain adenine biosynthetic mutations in the yeast Saccharomyces cerevisiae results in a red colony color. This phenomenon has historically provided an ideal genetic marker for the study of mutation, recombination, and aneuploidy in lower eukaryotes by classical genetic analysis. In this paper, it is reported that cells carrying ade1 and/or ade2 mutations exhibit primary fluorescence. Based on this observation, the nonselective enrichment of yeast cultures for viable adenine mutants by using the fluorescence activated cell sorter has been achieved. The advantages of this approach over conventional genetic analysis of mutation, recombination, and mitotic chromosomal stability include speed and accuracy in acquiring data for large numbers of clones. By using appropriate strains, the cell sorter has been used for the isolation of both forward mutations and chromosomal loss events in S. cerevisiae. The resolving power of this system and its noninvasiveness can easily be extended to more complex organisms, including mammalian cells, in which analogous metabolic mutants are available. PMID- 3044712 TI - Condensation of DNA in situ in metaphase chromosomes induced by intercalating ligands and its relationship to chromosome banding. AB - Interactions of certain intercalating cationic ligands with nucleic acids result in the formation of products that undergo condensation and agglomeration; this transition in solution can be monitored by light-scatter measurements. In the present study, using such intercalators as the antitumor drug mitoxantrone or fluorochromes acridine orange and quinacrine, we induced condensation of DNA in situ in Chinese hamster chromosomes. The in situ products scattered light and could be detected by darkfield- or phase-contrast microscopy. In the darkfield the complexes had a characteristic granular appearance and often generated a banding pattern on the chromosomes. In contrast, condensation of DNA in situ by the nonintercalating polyvalent cations (Co3+, spermine4+), while enhancing the chromosome's image contrast, did not produce the granular products or the banding. The condensation of free DNA, single or double stranded, natural or synthetic, the latter of various base composition and configuration, was also measured in solution. The condensation in solution and in situ was observed at similar concentrations of the respective ligands. The intercalating dye ethidium bromide, which did not condense DNA in solutions of moderate and high ionic strength, also did not generate the granular products or banding on chromosomes. The data also show that both base composition and configuration are important factors in determining the sensitivity of DNA to condensation by particular intercalating ligands. The studies suggest that the phenomenon of DNA condensation by intercalating dyes, which shows a high degree of specificity with respect to primary and secondary structures of DNA, may be associated with mechanisms of chromosome banding induced by the intercalating thiazine dyes in Giemsa staining or by quinacrine. Observation of chromosome banding based on light-scatter detection in darkfield microscopy allows the study of interactions between DNA and the ligands that neither fluoresce nor generate colored products. This principle of chromosome "counter-staining" can be explored by flow cytometry. PMID- 3044713 TI - Bone changes in alcoholic liver cirrhosis. A histomorphometrical analysis of 52 cases. AB - Bone biopsies of 52 histologically confirmed alcoholic cirrhotic patients and 15 age- and sex-matched controls have been histomorphometrically analyzed determining trabecular bone volume (TBV), mineralized bone volume (MBV), and osteoid volume (OV). We also determined serum PTH, 25-OH-D3, calcitonin, FSH, LH, estradiol, testosterone, T3 and T4, urine cortisol, routine liver function tests, serum and urinary calcium, phosphorus, and magnesium. We found a high prevalence of osteoporosis: TBV was significantly lower in cirrhotic patients (T = 7.23, P less than 0.001), 41 of them being in the range of osteoporosis; none of them had osteomalacia. Levels of all the above-mentioned hormones and electrolytes were almost normal, and no correlation was found between them and liver function tests, as occurred with the bone parameters. PMID- 3044714 TI - Unconjugated bilirubin in hepatic bile with brown pigment gallstones and cholangitis. AB - To investigate the role of cholangitis in hydrolysis of bilirubin in bile with brown pigment gallstones, bilirubin composition and bacterial growth in hepatic bile with and without cholangitis were studied. The study included 38 brown pigment gallstone cases (28 without cholangitis and 10 with cholangitis). The proportion of unconjugated bilirubin in hepatic bile with cholangitis (16.9 +/- 8.5%, mean +/- SD) was significantly higher than that without cholangitis (3.7 +/ 1.8%, P less than 0.001). A positive correlation was found between bacterial population with beta-glucuronidase activity and the proportion of unconjugated bilirubin in bile in cases of brown pigment stones with cholangitis (P less than 0.05) but not in those without cholangitis despite the fact that bacterial species and population are similar regardless of the presence of cholangitis. In cholangitis, pH of bile becomes lower toward the optimal pH of bacterial beta glucuronidase. Together the lower concentration of bile acid and the lower pH in bile result in lower solubility of unconjugated bilirubin, promoting its precipitation. Thus occasional bouts of cholangitis may result in periodic deposition of bilirubinate on brown pigment stones with layered structures by inducing cyclic changes of bile composition in situ. PMID- 3044715 TI - Ambulatory 24-hour esophageal pH monitoring. Reproducibility and variability of pH parameters. AB - If 24-hour esophageal pH monitoring is to be a useful diagnostic tool, it must reliably discriminate gastroesophageal reflux patients despite daily variations in distal esophageal acid exposure. To address this issue, we studied 53 subjects (14 healthy normals, 14 esophagitis patients, and 25 patients with atypical symptoms) with two ambulatory pH tests performed within 10 days of each other. Intrasubject reproducibility of 12 pH parameters to discriminate the presence of abnormal acid reflux was determined. As a group, the parameters of percent time with pH less than 4 (total, upright, recumbent) were most reproducible (80%). Therefore, a subject was defined as having gastroesophageal reflux disease if at least one of these three values were abnormal. Intrasubject reproducibility for the diagnosis of reflux disease was 89% for the entire sample. Among subsets, the reproducibility was 93% for the normals and esophagitis patients and 84% for the atypical symptom patients. Total percent time with pH less than 4 was the single most discriminate pH parameter (85%) and nearly equaled that of the three combined parameters (89%). The intrasubject variability of this parameter was determined by the mean +/- 2 SD of the relative differences between the two test results for all 53 subjects. Total percent time with pH less than 4 may vary between tests by a factor of 3.2-fold or less (218% higher to 69% lower). We conclude: (1) ambulatory 24-hr esophageal monitoring is a reproducible test for the diagnosis of gastroesophageal reflux disease; and (2) the large intrastudy variability in 24-hr total acid exposure may limit this test's usefulness as a measurement of therapeutic improvement. PMID- 3044716 TI - Adenomatous and hyperplastic polyps cannot be reliably distinguished by their appearance through the fiberoptic sigmoidoscope. AB - This prospective study is designed to determine if experienced sigmoidoscopists can, from gross appearance, differentiate adenomas and hyperplastic polyps. Six hundred eleven polyps discovered at fiberoptic sigmoidoscopy were removed completely or biopsied for histologic examination. The polyp's size, distance from the anal verge, color, and the endoscopist's diagnosis were analyzed. Hyperplastic polyps were significantly smaller, of lighter color, and located closer to the anus than adenomas. The sensitivity of the endoscopic diagnosis for adenomas is 0.80 and the specificity 0.71. We conclude the endoscopic diagnosis of polyps 1.0 cm and smaller is not accurate enough for decision making, and these should be biopsied to guide management. Since 97% of polyps larger than 1.0 cm are adenomas, their removal can be advised without biopsy. PMID- 3044717 TI - Fulminant hepatitis due to reactivation of chronic hepatitis B virus infection after allogeneic bone marrow transplantation. AB - A case of hepatitis B reactivation following bone-marrow transplantation for leukemia in a previously healthy HBsAg carrier is reported. A number of changes in HBV serum markers were contemporary to the acute episode. All of them (increase of HBsAg concentration, conversion from anti-HBe to HBeAg, appearance of anti-HBc IgM and of serum HBV-DNA) were suggestive of a "switching-on" of viral replication. Institution of corticosteroid treatment at the onset of the acute phase did not prevent the fatal outcome. PMID- 3044719 TI - Congestive heart failure. AB - Congestive heart failure (CHF) is not a single entity but a symptom complex that may represent the consequence of mechanical abnormalities, myocardial abnormalities, and/or disturbances of cardiac rhythm. In turn, it affects virtually every organ system in the body. This review focuses on CHF due to systolic dysfunction of the left ventricle, which comprises the majority of cases of this condition. Recent data suggest that CHF may be the most frequent primary diagnosis in patients on medical services in nonmilitary hospitals in this country: it affects approximately 2% of the United States population, or some 4 million people. The mortality rate for CHF is also worse than for many forms of cancer; thus, new therapeutic alternatives are imperative. In order to devise new therapeutic strategies, a detailed understanding of the pathophysiology of this condition is required. The relative advantages and disadvantages of various pharmacologic and nonpharmacologic approaches are considered in detail. Certain medications, such as the angiotensin converting enzyme (ACE) inhibitors, have been shown to improve survival, and heart transplantation is clearly life-saving for those who are eligible for this therapy. However, the real challenge is to devise strategies to prevent the occurrence of heart failure, or interrupt its progress at a very early stage. PMID- 3044718 TI - Endocrinology in aging. AB - Aging is a time of reduced adaptability to metabolic perturbation. This is particularly true in endocrinology which, after all, is the science of chemically regulated biologic systems. There is no evidence that equilibrium concentrations of the principal hormones are altered with age. However, the systems utilized to reach those equilibria become progressively taxed, and new equilibria may be achieved reflecting that regulatory problem. Thus, with advancing age there are significant alterations in hormone production, metabolism, and action. Some of these changes may play a role in the pathophysiology of senescence, although the evidence for that is limited. The magnitude of age-related alterations is highly variable and sex dependent. Whereas only subtle changes occur in pituitary dynamics, adrenal gland physiology, and thyroid function, the changes in glucose homeostasis, reproductive function, and calcium metabolism are more apparent. In the elderly, the interpretation of endocrine tests should reflect the nutritional status of the patient and the presence of coexisting illnesses. In this review, we describe the principles of endocrinology in the geriatric population and elaborate on the changes in specific glandular functions with aging. We also define strategies of evaluation and management protocols appropriate for the elderly with suspected endocrine dysfunction. PMID- 3044720 TI - [Endosonographic findings of benign and malignant lesions in the stomach wall. A prospective comparison with conventional imaging study procedures]. AB - The results of endoscopic ultrasonography in the diagnosis and differentiation of benign and malignant lesions of the stomach was prospectively compared with those obtained by upper endoscopy with biopsy, double-contrast radiography and computed tomography in 15 patients with benign and 26 with malignant gastric lesions. The validity of the methods was established by comparing the results obtained with the long-term course observed clinically in conjunction with an endoscopic histological follow-up or with the findings on histopathological examination of the operative specimen. Precise diagnosis of the lesion was made by endoscopic ultrasonography in about 90% of cases, a result better than that obtained with radiography or computed tomography. Furthermore, endoscopic ultrasonography had good sensitivity for demonstrating perigastric lymph node enlargement; in this respect it was better than computed tomography, but was not able to distinguish whether the enlargement was benign or malignant: this will only become possible with technical improvement of the instruments. PMID- 3044721 TI - [Visual disorders in the vertebrobasilar insufficiency syndrome]. PMID- 3044722 TI - [Renoparenchymal hypertension. Is there a specific therapy?]. PMID- 3044724 TI - [The early breakfast--an unknown pitfall in the treatment of type-I diabetes mellitus]. AB - In 15 type I diabetics the mean postprandial rise in blood glucose levels, 90 min after breakfast at about 8 a.m. (the customary time for this meal in hospitals), was 62.2 +/- 34.3 mg/dl, but 90 min after a breakfast which had been put forward by about 1 1/2 hours it was only 2.6 +/- 18.6 mg/dl (P less than 0.001). Seven patients developed hypoglycaemia after the early breakfast, none after the late one. This finding suggests that, to avoid hypoglycaemia after an early breakfast, an additional small meal should be given at about 8:30 a.m. or the insulin content of the morning injection should be changed. The clear connection between the time of the morning insulin injection and the insulin action during the morning explains why, in uninstructed patients who--under their usual daily conditions of starting school or work--are forced to inject insulin early or to eat early, a state of hypoglycaemia may occur. PMID- 3044723 TI - [Emergency therapy of ventricular tachycardias: lidocaine versus ajmaline]. AB - The efficacy of ajmaline (50-75 mg i.v.) or lidocaine (100-200 mg i.v.) in terminating persistent, haemodynamically stable ventricular tachycardia (VT) was compared in a prospective, randomized trial of 31 patients. There were no significant differences as to age, underlying heart disease, ejection fraction and rate of ventricular tachycardia between the two treatment groups. Ajmaline terminated VT in 10 of the 15 patients receiving it, lidocaine in only 2 of 16 (P less than 0.01). The frequency of VT was not significantly changed by lidocaine, while mean cycle length during VT changed under ajmaline from 369 +/- 82 ms to 452 +/- 11 ms (P less than 0.01). In contrast to lidocaine, QRS duration under ajmaline lengthened from 166 +/- 18 ms to 200 ms +/- 28 ms (P less than 0.01), but return cycles after tachycardia termination were similar (ajmaline, 863 +/- 296 ms; lidocaine, 917 +/- 367 ms). Both drugs were equally well tolerated, but in this series ajmaline was more effective in the acute treatment of persistent VT. PMID- 3044725 TI - [Tetanus prophylactic inoculation--the indications and contraindications]. PMID- 3044726 TI - [An increase in the oral calcium provision--a benefit or a risk?]. PMID- 3044727 TI - [Contribution of positron emission tomography to the diagnosis of dementia]. PMID- 3044729 TI - [Confirmation studies and strategies of analysis]. PMID- 3044730 TI - [Application of the t-test to between-group comparisons]. PMID- 3044728 TI - [Synopsis of infectious pneumonia]. PMID- 3044731 TI - [Bone-free bone grafts using electrically stimulated periosteum. Preliminary report and clinical applications]. PMID- 3044732 TI - [Historical development of bone substitutes in the jaws]. PMID- 3044733 TI - [Formation of porosities in ceramo-veneered palladium-silver alloys]. PMID- 3044734 TI - [Borderline cases in conservative dentistry with respect to prosthodontic treatment]. PMID- 3044735 TI - [Experience with bonded bridgework. A multicenter observation study. 5. Success of rebonding and second treatment with new constructions]. PMID- 3044736 TI - [Combined surgical-prosthodontic treatment for maxillary hypoplasia. A case report]. PMID- 3044737 TI - [Ossifying fibroma of the maxilla--course, surgical treatment and prosthodontic aftercare]. PMID- 3044738 TI - [Extraoral photopolymerization for optimum physical-technical characteristics of composites]. PMID- 3044739 TI - [A new titanium post system for the combination of apicoectomy with a plastic core]. PMID- 3044740 TI - [Studies on mechanical stresses in porcelain fused to metal]. PMID- 3044741 TI - [In vitro corrosion tests of different alloy crowns over amalgam cores]. PMID- 3044742 TI - [Effects of cements on the corrosion of post and core crowns]. PMID- 3044743 TI - [Etching and silanizing of dental ceramics]. PMID- 3044744 TI - [Effects of a chrome-cobalt bonding agent on the bonding strength of ceramics on partial denture casting alloys]. PMID- 3044745 TI - [Bending strength tests of ceramo-metallic systems with different dental alloys]. PMID- 3044746 TI - [Silicoater method for bonded bridgework]. PMID- 3044748 TI - [UDA anchor system for replacement of missing teeth]. PMID- 3044747 TI - [Thermal expansion and plasticizing temperatures of dental adhesives]. PMID- 3044750 TI - [Easy construction technique for bridgework with Ultralite-foils]. PMID- 3044749 TI - [Accuracy of fit of castings in relation to impression, model and technical parameters]. PMID- 3044751 TI - [Contact area between male and female components of prefabricated attachments]. PMID- 3044752 TI - [Guiding characteristics of different attachments and clasps with respect to the kinematics of free-end saddles]. PMID- 3044753 TI - [Influence of manufacturing processes and surface treatment on the retentive power of taper crowns]. PMID- 3044754 TI - [Resistance of hinges to wear]. PMID- 3044755 TI - [Accuracy of fit of partial denture castings]. PMID- 3044756 TI - [Strength of the Kamba-bloc attachment]. PMID- 3044757 TI - [Clinical success of taper crowns as perioprosthodontic construction elements. A long-term study]. PMID- 3044758 TI - [Treating posterior edentulism with the aid of implant-anchored removable bridges]. PMID- 3044760 TI - [Effect of dimensions on the strength of crowns]. PMID- 3044759 TI - [Clinical long-term results with partial dentures with resilient support]. PMID- 3044761 TI - [Reproducibility of cast crowns depending on different wax modelling techniques]. PMID- 3044762 TI - [Immunocytochemical studies of the oral mucosa in hyperplastic conditions caused by irritation]. PMID- 3044763 TI - [Hybridization with synthetic oligonucleotide probes: a new procedure for fast bacteriological diagnosis of inflammatory periodontal diseases]. PMID- 3044766 TI - [Effects of periodontal instruments on the root surface and its bacterial colonization]. PMID- 3044765 TI - [A new approach to furcation treatment]. PMID- 3044764 TI - [Evaluation of molecular biological probes for the direct identification of complex bacteria in inflammatory periodontal diseases]. PMID- 3044767 TI - [Epithelial reattachment following root surface treatment using different instruments]. PMID- 3044768 TI - [Root-resected molars as abutment teeth]. PMID- 3044769 TI - [Ultrasonic imaging of periodontal structures--course and results of a research project]. PMID- 3044770 TI - [Methods for creating hyperthermia in tumors by using electromagnetic fields]. AB - Evidence on the methods for creation of hyperthermia in treatment of human tumours using superhigh-frequency external, intracavitary, and implanted irradiators are generalized. Inductive, capacitive and interstitial methods of heating are described. Advantages and shortcomings, depths of heating and heat production structures are presented for each method. The known devices for regional hyperthermia are considered. PMID- 3044771 TI - [Mechanisms of metastasis and factors in antimetastatic resistance]. AB - The present notions of pathogenetic mechanisms of metastatic spreading are discussed, and results of the investigations devoted to an increase of antimetastatic resistance of the body, application of different immunomodulators, neurotropic and antiangiogenic influences during surgery, chemotherapy, and irradiation, as well as individualization of antimetastatic therapy aimed at prevention, diagnosis and treatment of metastases of human malignant tumours are described. PMID- 3044772 TI - [Modern approaches to the molecular diagnosis of leukemia]. AB - New outlooks in the diagnosis of leukemia and lymphomas with using recombinant DNA technology are reported. Possibility of DNA and RNA analysis to detect specific gene markers are demonstrated. It is a new tool for the precise identification of malignant clones of hematopoietic cells and for the improvement of early diagnosis. PMID- 3044773 TI - [Further comment on the terminology of carcinogenesis and mechanism of action of chemical carcinogens]. PMID- 3044774 TI - [Results of a discussion on the terminology of carcinogenesis]. PMID- 3044775 TI - [Modifying effect of environmental chemical factors on the metabolic activation of carcinogens]. AB - Current investigations of modifications of metabolic activation of carcinogens by the environmental chemical factors which are able to induce or inhibit the metabolic activation are reviewed. From the standpoint of ecological oncology the most promising are the following trends: 1) modelling under experimental conditions of integral ecosystems including the complex of living organisms, environmental carcinogens, and factors which modify their biotransformation; 2) study of the imprinting effectors of metabolism modifiers; 3) detection in the environment of modifiers of metabolic activation of carcinogens. The comprehensive oncological-ecological studies of chemicals which influence different pathways of carcinogen metabolism are very important for both primary prevention of cancer and the environmental protection. PMID- 3044776 TI - Reproductive endocrine disorders in women with primary generalized epilepsy. AB - It is known that women suffering from temporal lobe epilepsy may frequently present reproductive endocrine disorders (REDs). We hypothesized that a high occurrence of REDs could be found also in primary generalized epilepsy (PGE), and therefore investigated the hormonal and ovarian echographic profiles in 20 PGE female patients of reproductive age. Fourteen reported normal menstrual cycles, while 6 complained of longstanding menstrual irregularities. All but three patients were receiving antiepileptic drug (AED) therapy. In all subjects, the basal levels of gonadotropins, prolactin, and gonadal steroids were assayed. The response of luteinizing hormone (LH) to gonadotropin-releasing hormone was also investigated and ovarian ultrasonographic findings were evaluated. In five of six patients with menstrual problems (25% of the group), a well-defined RED was diagnosed (polycystic ovarian disease in three cases and hypothalamic ovarian failure in two). The 14 patients with normal menstrual cycles showed an elevation of mean basal follicle-stimulating hormone and prolactin, and a blunting of mean LH response. Our results suggest that a high occurrence of REDs may be found also in PGE. We hypothesize that a neurotransmitter dysfunction might be the common pathogenetic mechanism resulting in both REDs and PGE. The hormonal alterations observed in the patients with normal menstrual cycles seem to support our hypothesis. Previous data seem to rule out a possible AED effect accounting for the hormonal findings observed in our series. However, further studies are needed to confirm our preliminary results. PMID- 3044777 TI - Increased insulin receptors in carbohydrate-sensitive subjects: a mechanism for hyperlipaemia in these subjects? AB - Twenty male subjects, 11 normal and 9 carbohydrate-sensitive, participated in a study in which the effect of feeding diets low in copper (1.03 mg/d) on the number and affinity of insulin receptors was determined. Since carbohydrate sensitive subjects are hyperinsulinaemic, it was anticipated that they would demonstrate a down-regulation of insulin receptors. The subjects were fed a low copper diet for 11 weeks and then replenished with copper (3 mg/d) for 3 weeks. Regardless of diets fed, carbohydrate-sensitive subjects showed increased insulin binding. The increase was due to the number of receptors without any change in their affinity. This unusual and unexpected observation in carbohydrate-sensitive subjects suggests an altered response of the insulin receptor, namely the failure of plasma insulin to down-regulate the number of receptors. It is possible that such an alteration could lead to a shift of the biological response curve of insulin to the left in some target tissues thereby causing hyperlipaemia by diverting glucose into triglyceride. The exact mechanism of hyperlipaemia in carbohydrate-sensitive subjects remains to be clarified. PMID- 3044778 TI - Light-dependent changes in psbD and psbC transcripts of barley chloroplasts: accumulation of two transcripts maintains psbD and psbC translation capability in mature chloroplasts. AB - The psbD and psbC genes encode two polypeptides of Photosystem II. These genes are adjacent in the barley chloroplast genome and are part of a 5.7 kbp transcription unit. In dark-grown barley, four large transcripts hybridize to psbD and psbC; two additional transcripts hybridize to psbC. Illumination of 4.5 day-old dark-grown seedlings causes a decrease in the six psbD--psbC transcripts found in etioplasts and the accumulation of two different transcripts of 4.0 and 3.2 kb which hybridize to psbD and psbC. The light-induced transcripts have a common 5' end approximately 600 nt upstream of psbD and 3' ends 1175 and 175 nt downstream of psbC. The shift in psbD--psbC transcript population occurs during a phase of chloroplast maturation when transcript levels and translation of chloroplast genes such as psaA--psaB and psbB decline approximately 3- to 5-fold. In contrast, translation of the psbD and psbC gene products declines to a lesser extent, suggesting that the light-induced accumulation of the 4.0 and 3.2 kb psbD -psbC transcripts is required to maintain psbD and psbC gene product translation in mature chloroplasts. PMID- 3044779 TI - Amino acid sequence of the murine Mac-1 alpha chain reveals homology with the integrin family and an additional domain related to von Willebrand factor. AB - Clones encoding the Mac-1 alpha chain were selected from a mouse macrophage cDNA library by screening with oligonucleotide probes based on the sequence of a genomic clone encoding the N-terminus of the mature protein. The sequence of overlapping clones (4282 nt) was determined and translated into a protein of 1137 amino acids and a signal peptide of 15 amino acids. The Mac-1 sequence was found to be related to the alpha chain sequences of three other members of the integrin family of cell adhesion receptors, i.e. the fibroblast receptors for fibronectin and vitronectin and the platelet glycoprotein IIb/IIIa. All four sequences share a number of structural features, like the position of 13 cysteine residues, a transmembrane domain near the C-terminus and the location of three putative binding sites for divalent cations. Furthermore, a conserved sequence motif is repeated seven times in the N-terminal half of the molecule and three of these repeats include putative Ca/Mg-binding sites of the general structure DXDXDGXXD. On the other hand, Mac-1 contains a unique domain of 220 amino acids inserted into the N-terminal part of the integrin structure. This additional domain is homologous to a repeated domain found in von Willebrand factor, cartilage matrix protein and in the complement factors B and C2. In two of these proteins, the homologous domain is likely to be involved in binding to collagen fibrils. Therefore, Mac-1 may also bind to collagen, which could play a role in the interaction of leukocytes with the subendothelial matrix. PMID- 3044780 TI - MAS1, a gene essential for yeast mitochondrial assembly, encodes a subunit of the mitochondrial processing protease. AB - We have previously described a yeast mutant (mas1) that accumulates mitochondrial precursor proteins at high temperature and is deficient in the activity of a matrix-localized protease which cleaves presequences from mitochondrial precursor proteins. We have now cloned and sequenced the wild-type MAS1 gene and found that it encodes a subunit of the mitochondrial processing protease, that it is essential for cell viability and that the protein product participates in its own cleavage during import into mitochondria. The MAS1 protein is thus the first genetically defined component of the mitochondrial protein import pathway. PMID- 3044781 TI - Secretion of invertase in mitotic yeast cells. AB - In mammalian cells intracellular transport is inhibited during mitosis. Here we show that in the yeast Saccharomyces cerevisiae secretion continues uninterrupted during mitosis. S. cerevisiae cells were arrested in mitosis by treating wild type cells with the microtubule-inhibitor nocodazole, or by incubating a temperature-sensitive cell division cycle mutant (cdc16) at the restrictive temperature. Secretion of invertase into the periplasmic space was equally efficient in mitotic and in unsynchronized cells. Electron microscopy of nocodazole-treated mitotic wild-type cells revealed stretches of rough endoplasmic reticulum, strongly fenestrated Golgi cisternae and clusters of vesicles with the diameter of 30-90 nm. Secretion of invertase was inhibited in mitotic sec7 cells at the restrictive temperature, but continued at the permissive temperature. Sec7 is a mutant strain where intracellular traffic is blocked in unsynchronized cells in the Golgi complex at the restrictive temperature. Thus, the elements of the mitotic Golgi complex appear to be able to support intracellular traffic. PMID- 3044783 TI - Selective degradation of mRNA: the role of short-lived proteins in differential destabilization of insulin-induced creatine phosphokinase and myosin heavy chain mRNAs during rat skeletal muscle L6 cell differentiation. AB - This investigation concerns the combined effects of removal and readdition of insulin and inhibition of protein and RNA synthesis on the stability of insulin induced mRNAs during and after differentiation of rat L6A1 myoblast cells in culture. Addition of insulin accompanying the withdrawal of the mitogenic stimulus of serum to myoblasts caused an 80-fold increase in creatine phosphokinase (CK) activity which was largely accounted for by a similar increase in the amount of CK mRNA. The latter was co-ordinately induced with myosin heavy chain (MHC) mRNA but not malic enzyme (ME) mRNA. Measurements of steady-state levels of mRNA showed that removal of insulin caused CK mRNA, but not MHC mRNA, to be rapidly degraded, the effect being reversed upon readdition of the hormone. Direct measurement of 3H-labeled CK, MHC and beta-actin mRNAs confirmed the selective stabilization and destabilization of CK mRNA by the hormone. Conditions were established for a time-window during which cycloheximide (Cx) produced a virtually total arrest of protein synthesis in myotubes that was reversible upon removal of the inhibitor. Under these conditions, Cx selectively prevented the degradation of CK mRNA in a reversible manner. Actinomycin D (Act D) also arrested the loss of this mRNA. Under the same conditions of mRNA stabilization during de-induction, a superinduction of CK mRNA, but not MHC mRNA, was observed if the two inhibitors were added during induction in the continuous presence of insulin. We conclude that a short-lived protein(s), encoded by a short-lived mRNA(s), selectively regulates the stability of reversibly inducible mRNA. PMID- 3044782 TI - Purification and characterization of the inducible a agglutinin of Saccharomyces cerevisiae. AB - A cell surface glycoprotein induced by the mating pheromone alpha factor in Saccharomyces cerevisiae a cells has been purified to homogeneity. At 4 x 10(-9) M it strongly inhibits mating-type-specific agglutination between a and alpha cells. The protein is solely O-glycosylated. It consists of 29% carbohydrate and its apparent molecular mass is 22 kd on SDS gels. After HF treatment it behaves like a protein of 13 kd; therefore its true molecular mass probably is close to 18 kd. Mild periodate treatment destroys the biological activity of the purified protein. The protein contains one cysteine, no arginine, and 27% of the amino acids are serine and threonine residues, two thirds of which are glycosylated. With a polyclonal antibody the glycoprotein can already be detected at the cell surface 15 min after pheromone addition. The inducible antigen is not expressed in a specific phase of the cell cycle; it first appears exclusively on the growing bud. Mother cells express the antigen on their surface only after the daughter cells have separated; it is then localized at the tip of the pear-shaped 'shmoo'. Using the secretory ts-mutant sec 18 is shown that a mannosylated precursor of a agglutinin accumulates at the endoplasmic reticulum. PMID- 3044784 TI - Mutagenic screening of marker grenade dyes by the Salmonella reversion assay, L5178Y/TK+/- mouse lymphoma assay, and in vivo sister chromatid exchange analysis in mice. AB - Two dyes (C.I. Solvent Yellow No. 33 and a mixture of C.I. Solvent Yellow No. 33 and C.I. Solvent Green No. 3) were tested for mutagenicity in the Salmonella reversion assay and the L5178Y/TK+/- mouse lymphoma assay, and also for sister chromatid exchange (SCE) induction in vivo in C57B1/6J mice. In addition, a greater than 99.9% pure sample of the yellow dye [2-(2'-quinolyl)-1,3-indandione] was tested with and without exogenous activation in the Salmonella reversion assay and the L5178Y/TK+/- mouse lymphoma assay. Neither C.I. Solvent Yellow No. 33 nor the C.I. Solvent Yellow No. 33 and Solvent Green No. 3 mixture was positive for inducing SCEs in vivo. All three dyes were tested in the standard plate incorporation test in seven Salmonella strains TA98, TA100, TA102, TA104, TA1535, TA1537, and TA1538. The dyes were negative with and without exogenous activation in TA98, TA1535, and TA1538. One test with TA1537 was positive with the greater than 99.9% purified yellow dye. All three dyes gave weakly positive results (less than a twofold increase) with S-9 in TA100 and were clearly positive in TA102 and TA104 both with and without S-9. They also induced mutation at the thymidine kinase locus in mouse lymphoma cells, produced both large- and small-colony trifluorothymidine-resistant mutants, and were clastogenic. The purified yellow dye was further tested for SCE induction in mouse lymphoma cells and was determined to give a slightly positive response in the presence of S-9. PMID- 3044785 TI - Weak and unexpected mutagenicity to Salmonella of the rat hepatocarcinogen methapyrilene. AB - The rat liver carcinogen methapyrilene is shown to be a selective mutagen to strain TA1535 of Salmonella typhimurium when tested in the absence of S9 mix and using the standard plate-incorporation assay protocol. The activity observed was weak but was reproducible for a range of samples on many occasions of test and was not due to impurities. These data contrast with six earlier reports of the inactivity of this chemical in the Salmonella mutation assay. PMID- 3044786 TI - Mutagenicity of halogenated propanes and their methylated derivatives. AB - 1,2,3-Tribromopropane, 1,2,3-trichloropropane, and 1,2-dibromo-3-chloropropane are mutagenic in strains TA1535 and TA100 of Salmonella typhimurium, but only in the presence of rat liver S9 mix. This requirement for metabolic activation was unexpected for an alkyl halide and thus suggested the metabolic formation of the 2-keto derivatives (di-haloacetone). The 2-methyl derivatives of the halopropane compounds did not induce a doubling of revertants compared to controls. It was demonstrated that none of these compounds is converted into a secondary material that could be determined as structurally different by gas chromatography. These observations suggest that lack of mutagenicity of the methylated compounds is a manifestation of a steric effect. PMID- 3044787 TI - Synergistic killing of bacteria and phage by polystyrene and ultraviolet radiation. PMID- 3044788 TI - Only one out of the three strong ribosomal binding sites of the early region of bacteriophage T7 exhibits high translational efficiency in fragments of about 30 base pairs. AB - Within the early region of bacteriophage T7 three genes, 0.3, 1 and 1.3, are most efficiently expressed. They belong to the strongest initiation signals of Escherichia coli. In the T7 wild-type situation the proteins are produced with a molar ratio of gene 1:1.3:0.3 protein = 1:3.9:9.7. DNA fragments of about 30 base pairs comprising the ribosomal binding sites (RBS) of these genes were synthesized and cloned into derivatives of the pDS1 vector ribosomal binding sites (RBS) of these genes were synthesized and cloned into two derivatives of the pDS1 vector just upstream of the mouse dihydrofolate reductase gene. Although all tested RBS fragments contained an initiation triplet, a Shine-Dalgarno sequence and some nucleotides upstream and downstream of this region, only the gene 1.3 RBS fragment showed high efficiency whereas those of genes 0.3 and 1 were at the border of significance. The amount of synthesized mRNA was about the same for all three constructs. A major influence of vector-derived sequences on the RBS activity could be ruled out. The high translational activity of the short 1.3 gene RBS seems to be largely due to its primary structure. The other two RBSs studied require much longer sequences for high activity. PMID- 3044789 TI - Structural requirements of lipid A species in activation of clotting enzymes from the horseshoe crab, and the human complement cascade. AB - The structure/activity relationship of lipid A, a bioactive center of endotoxic lipopolysaccharides, in the activation of the clotting enzyme cascade of a horseshoe crab amoebocyte lysate (Limulus activity) and the complement system in human serum, was examined using synthetic lipids A and related compounds. Regarding Limulus activity, a newly developed colorimetric method, which utilizes a mixture of recombined clotting factors and a chromogenic substance, was much more sensitive for detecting changes in the chemical structure of test compounds than the conventional gelation method using the amoebocyte whole lysate. (beta 1 6)-D-Glucosamine disaccharide bisphosphates, which had neither 3-hydroxyacyl nor 3-acyloxyacyl groups, and acylglucosamine phosphates, which in structure correspond or are analogous to the non-reducing or reducing moieties of lipids A and biosynthetic disaccharide lipid A precursors showed only negligible activity in the colorimetric tests, but they exhibited a distinct though much weaker gelation activity than the parent disaccharide molecules. The assay results obtained by the colorimetric Limulus test correlate better with the pyrogenicity of the test synthetic compounds than those given by the gelation method, although the dependence of pyrogenicity on chemical structure is greater. The presence of 3-hydroxyacyl groups on the bisphosphorylated (beta 1-6)-D-glucosamine disaccharide backbone is the prerequisite for effective activation of the clotting enzyme cascade of horseshoe crab amoebocyte lysate, while the presence of an adequate number (one or two) of 3-acyloxyacyl groups on the disaccharide bisphosphate backbone is needed for full pyrogenicity. Complement activation, on the other hand, showed structural requirements quite different from those for the colorimetric Limulus activity and the pyrogenicity. The disaccharide compounds that had only non-hydroxylated acyl groups, acylated glucosamine phosphates that had the structure of the non-reducing portion of lipids A and biosynthetic disaccharide precursors, which were scarcely active in the colorimetric Limulus test, caused complement activation comparable to or sometimes stronger than that of the parent disaccharide molecules. Acylglucosamine phosphates, corresponding in structure to the reducing moiety of disaccharide compounds, however, showed little activity. PMID- 3044790 TI - Hyper-regulation of pyr gene expression in Escherichia coli cells with slow ribosomes. Evidence for RNA polymerase pausing in vivo? AB - UTP-modulated attenuation of transcription is involved in regulating the synthesis of pyrimidine nucleotides in Escherichia coli. Thus, expression of two genes, pyrBI and pyrE, was shown to be under this type of control. The genes encode the two subunits of aspartate transcarbamylase and orotate phosphoribosyltransferase respectively. The levels of these enzymes are inversely correlated with the intracellular concentration of UTP. Modulation of attenuation seems to be a consequence of the effect of UTP concentration on the mRNA chain growth rate. Reducing the UTP pool retards RNA polymerase movement. Mechanistically this will couple the ribosomes translating a leader peptide gene more tightly to the elongating RNA polymerase. The ribosomes will then be more prone to prevent the folding of the mRNA chains into terminating hairpin structures when RNA polymerase is at the attenuator and has to decide whether transcription should terminate or continue into the structural genes. This paper described a study of pyrBI and pyrE gene regulation in cells where the ribosomes move slowly as a result of mutation in rpsL. It appears that expression of the two genes is hyper-regulated by the UTP pool in this type of cells. Furthermore, the attenuator model can only account for the results if it is assumed that UTP concentration-dependent pausing of transcription occurs in vivo in the two pyr gene leaders such that RNA polymerase waits for the coupled ribosomes before transcribing into the attenuator regions. PMID- 3044791 TI - Congestive heart failure: how to evaluate efficacy of treatment. AB - Vasodilators are widely used in the therapy of congestive heart failure. Different mechanisms responsible for a failure of vasodilator therapy are discussed. Tolerance and/or diminished reabsorption or a less than optimal choice of the drug may be responsible for the lack of acute response. In addition, counter-regulation, mainly by the sympathetic nervous system, may decrease the drug effect. Furthermore, the initial beneficial effect of each vasodilator can be diminished with long-term therapy. Based on a clinical assessment and on invasive and noninvasive techniques a proposal is made as to how to treat an individual patient efficaciously. PMID- 3044793 TI - Effect of vasodilator therapy on mortality in chronic congestive heart failure. AB - The reduction of mortality in patients with chronic congestive heart failure treated with the vasodilator regimen hydralazine and isosorbide dinitrate compared to those treated with placebo or prazosin in the Veterans Administration Cooperative Study (V-HeFT) was examined in order to explore the possible mechanism of the favourable effect. Similar efficacy in coronary and non-coronary disease patients tends to discount a prominent effect on myocardial perfusion. The most likely explanation for the prolonged survival on the effective vasodilator regimen is that these drugs tend to increase left ventricular ejection fraction, probably by a sustained effect on preload and impedance. PMID- 3044792 TI - On biochemical and physiological indicators of stress relevant to cardiovascular illness. PMID- 3044794 TI - Does echocardiography allow the monitoring of the cardiac effects of nitrates? AB - Echocardiography is a very useful non-invasive method of cardiac diagnosis. It has been widely used in evaluating the effects of some drugs, such as nitrates, prazosin, and propranolol, on heart and seems very successful. In this paper, the echocardiographic studies of the effects of nitrates on left ventricular dimension and volume have been reviewed and an attempt has been made to answer whether or not echocardiography can allow to monitor the effects of nitrates in patient groups and further in individual patients. PMID- 3044797 TI - Infantile cortical hyperostosis associated with the Wiskott-Aldrich syndrome. AB - A case report of an infant with the Wiskott-Aldrich syndrome and clinical and radiological features of infantile cortical hyperostosis (Caffey disease) is presented. This is the third case described of the association of these two rare disorders and gives further support to the role of an immunologic defect in the pathogenesis of infantile cortical hyperostosis. PMID- 3044795 TI - Wiedemann-Beckwith syndrome. AB - The Wiedemann-Beckwith syndrome (WBS) comprises an accumulation of multiple congenital anomalies. Exomphalos, macroglossia and gigantism are considered the most common manifestations, hence the alternative designation EMG-syndrome. The syndrome carries with it an increased risk of developing specific tumours. One of the more frequent metabolic changes is transient neonatal hypoglycaemia, the result of increased insulin secretion. Inheritance of the syndrome remains uncertain. Most cases are sporadic, but a number of familial cases have been reported. Present evidence suggests that WBS is an autosomal dominant trait with variable expressivity. This review summarizes the abundant literature on the subject and discusses recent molecular genetic developments that may explain the interrelationship between the clinical abnormalities, metabolic disturbances and development of tumours. PMID- 3044796 TI - The DiGeorge syndrome. I. Clinical evaluation and course of partial and complete forms of the syndrome. AB - This study describes clinical signs and symptoms in 16 patients with the DiGeorge syndrome (DGS). Diagnosed on the basis of typical facial stigmata, a broad spectrum of severity is seen with respect to congenital heart disease, hypoparathyroidism and immunologic parameters. A simple index of severity is introduced that clearly differentiates complete forms of the syndrome (cDGS) with poor prognosis from partial forms of the syndrome (pDGS). Of 13 pDGS patients, 12 are still living; 8 underwent corrective heart surgery without infectious complications. Moderate to severe mental retardation is seen in all pDGS patients. Due to the lack of thymus function, immunodeficiency is a result of cDGS, whereas immunoregulatory disturbances (hypergammaglobulinaemia, high titres of specific antibody production) prevail in pDGS patients. PMID- 3044798 TI - On the efficacy of various irradiation regimens in phototherapy of neonatal hyperbilirubinaemia. AB - The efficacy of various irradiation regimens in phototherapy of neonatal hyperbilirubinaemia was analysed. One hundred and one newborns were assigned to three groups at random. The best results were achieved when six special blue fluorescent lamps were used and the sides of the incubator were draped with white cloth to reflect the light diffusely. This simple measure increased the therapeutic effect by approx. 35%. Compared to standard phototherapy units equipped with fluorescent lights, a halide lamp was no more effective. The results of the clinical trial confirm the conclusions drawn from measurements, published previously. According to those findings, irradiating large areas of skin as homogeneously as possible should produce optimum results. Using a lamp with a large surface in combination with diffusely reflecting areas should best meet this requirement. PMID- 3044799 TI - Intra-abdominal cystic lymphangioma in a newborn infant. Case report and review of the literature. AB - We report a newborn infant presenting with an intra-abdominal cystic lymphangioma, in which necrosis and infection led to an unusual combination of solid and liquid areas observed by ultrasonography. PMID- 3044800 TI - Induction and maintenance therapy in multiple myeloma: a multicenter trial of MP versus VCMP. AB - In a prospective multicenter trial, 320 untreated myeloma patients of stage II and III were randomized for remission induction into two groups receiving six monthly courses of either MP or VCMP treatment. Response rates were equal in both groups: 72% remission, 21% no change, 7% progress for patients evaluable by TCM changes and 56% remission, 11% no change, 33% progress for BJ- and non-secretory myelomas. The overall survival rate was 60% after 4 years. An unexpected finding was the significantly longer survival of MP treated patients compared to the VCMP group. After successful remission induction, patients were randomized into one group receiving maintenance treatment using the induction scheme q 8 weeks, and another group without further chemotherapy. Although patients in the latter group relapsed significantly earlier, differences between both groups concerning acquired resistance to first line therapy or survival have not been noticed to date. PMID- 3044801 TI - Nephrotoxicity of chemotherapy. PMID- 3044802 TI - Prolactin and breast cancer. PMID- 3044803 TI - Phase I study of a carboplatin-etoposide combination in advanced thoracic cancer. AB - We conducted a phase I trial with the combination carboplatin (CBDCA)-etoposide (VP-16) in thoracic cancer. CBDCA, at a starting dose of 300 mg/m2 dl, was associated with a fixed dose of VP-16 (100 mg/m2 dl-3). Escalation of doses was permitted after three patients entered at each dose level without grade IV toxicity. As expected, hematologic toxicity was the limiting factor. Severe myelosuppression (grade IV) occurred in three out of four patients treated at 350 mg/m2. Only three out of 19 patients treated at 325 mg/m2 exhibited a reversible grade IV hematologic toxicity. Other toxicities were mild and acceptable. Among 15 evaluable patients three showed a partial response. Two of the three responders have previously had a progression while receiving cisplatin and etoposide. The recommended dose of carboplatin to be associated with VP-16 (100 mg/m2 dl-3) is thus 325 mg/m2 dl. PMID- 3044804 TI - Endothelium-derived relaxing factor inhibits renin release. PMID- 3044805 TI - The endocrine pancreas of BB/OK-rats before and at diagnosis of hyperglycaemia. AB - From 148 BB-rats 76 animals (51.4%) developed hyperglycaemia within 55th to 178th day of life. Already before diagnosis of diabetes a great variety in pancreatic insulin content and morphometrically determined relative beta-cell volume density were visible. Despite of this great heterogeneity two types in the course of beta cell destruction could be observed. Verified by individual retrospective analysis one part of the rats is characterized by a more chronical course of diabetes development (pancreatic insulin content and beta-cell mass already more than 60 days before diagnosis of hyperglycaemia significantly reduced, occurrence of insulitis) whereas other animals show an acute form of beta-cell destruction (still about 20 days before diagnosis of diabetes pancreatic insulin content and relative beta-cell volume density in a normal range). Besides a drastical reduction of pancreatic insulin content at the time of diabetes diagnosis intense mononuclear infiltrations in the islets of Langerhans causing their destruction are demonstrable. As a result the residual beta-cell volume is significantly reduced. Rats with a plasma glucose higher than 13 mmol/l at diagnosis do not have any demonstrable beta-cells and an insulin content on the lowest detection limit of the method. In animals with a more mild hyperglycaemia (plasma glucose 8.3 to 13 mmol/l) there are rats with and the other ones without immunohistochemically detectable beta-cells. We conclude that prospective statements on the development of diabetes on the strength of analysis of residual beta-cells and pancreatic insulin content are not possible at present. We suppose that there exist different processes of beta-cell destruction which have to be investigated in further studies including immunological parameters. PMID- 3044806 TI - Complement-mediated cytotoxic effects of islet cell surface antibodies in non diabetic subjects with antecedent mumps infection and diabetic risk. AB - We studied sera of 24 selected non-diabetic subjects in whom an antecedent mumps infection with either complications and/or with onset at older age occurred 10-27 month ago regarding the capability to lyse rat islets of Langerhans in vitro. Thirteen subjects were characterized by diabetes associated HLA antigens DR3 and/or DR4 whereas 11 of them had none of these HLA antigens. Islet cell surface antibodies (ICSA) could be detected in 11 out of 24 subjects. Isolated pancreatic islets from 8-10 days old Wistar rats were incubated in freshly prepared human serum. The insulin leakage under the conditions of pharmacologic blockade of insulin secretion by means of epinephrine and propranolol provides a measure of complement-mediated cytotoxicity of human serum against rats islets in vitro. Out of a total of 24 subjects the sera of 11 of them exhibited cytotoxicity. Mean cytolytic insulin leakage was significantly higher in subjects with DR3 and/or DR4 in comparison with subjects without these HLA antigens (6.82 +/- 0.77% vs 3.90 +/- 0.48%; p less than 0.05). Ten out of the 11 subjects with cytotoxic sera had HLA DR3 and/or DR4 whereas DR3 was present in three out of 13 subjects with non-cytotoxic sera. There was no relationship between beta cell function in vivo (fasting C-peptide concentration and insulin response to oral glucose challenge) and the capability of patients sera to damage islet cells in vitro. In conclusion the pathogenetic role of mumps virus and cytotoxic ICSA and their possible relation to slow progressive beta-cell destruction has still to be ascertained. PMID- 3044808 TI - [Obituary of Prof. Emil Tonutti, M.D]. PMID- 3044807 TI - Effect of metoclopramide on orthostatic hypotension in diabetes mellitus: failure to demonstrate the role of vasoconstrictive hormones. AB - Effect of metoclopramide (MCP), a dopaminergic antagonist, on orthostatic hypotension associated with diabetes mellitus, was investigated to prove its plausible role of renin-angiotensin-aldosterone and catecholamine activation by comparative study in 6 diabetics with orthostatic hypotension (OH) and 9 diabetics without OH. With MCP treatment, drop of systolic blood pressure (SBP) on standing was significantly improved. However, the postural responses of plasma renin activity (PRA), plasma aldosterone and norepinephrine were not enhanced by MCP treatment. Moreover, MCP treatment did not modify the vascular reactivity to the exogenously infused angiotensin II and norepinephrine. Thus, it would be interesting to speculate that MCP can improve SBP through its suppressive effect on some depressor substance(s) such as bradykinin which liberates exaggeratively in diabetic OH. PMID- 3044809 TI - Histone hyperacetylation is induced in chick erythrocyte nuclei during reactivation in heterokaryons. AB - Transcriptionally inactive avian red blood cell nuclei were reactivated by Sendai virus-induced fusion of chicken erythrocytes with HeLa cells. We have used antibodies which specifically recognize the tetraacetylated form of H4 histone to show that histone hyperacetylation is an event required for chromatin reorganization leading to a transcriptionally competent chromatin structure. PMID- 3044810 TI - A developmental study of interphotoreceptor retinoid-binding protein (IRBP) in single and double homozygous rd and rds mutant mouse retinae. AB - Interphotoreceptor retinoid-binding protein (IRBP) was studied using immunochemical and immunocytochemical techniques in retinae of mice with allelic combinations at the rd and rds loci at different stages of development and degeneration. Until postnatal day 7 (P7), IRBP is located intracellularly in developing retinae of the different genotypes. Thereafter, IRBP is present mainly in the interphotoreceptor matrix. As previously noted, cell death is slowest in the heterozygous +/+,rds/+ mutant with loss increasing in order in +/+,rds/rds, rd/rd, rds/rds and rd/rd,+/+ animals. The IRBP content of the total retina also approximates this pattern, with lowest amounts by far in rd/rd, rds/rds and rd/rd,+/+ mutants (after P14). Interestingly though, IRBP loss significantly precedes visual cell loss in the rd/rd,rds/rds retina. In all the mutants, the remaining rod cells in the outer nuclear layer exhibit synthesis of intracellularly located IRBP at late stages of degeneration. In the single homozygous rd/rd,+/+ and the double homozygous rd/rd,rds/rds mutants, IRBP is present intracellularly during the entire degenerative process with somewhat less intracellular IRBP in the rd/rd,rds/rds mutant. Retinae of homozygous +/+,rds/rds and heterozygous +/+,rds/+ animals exhibit a normal distribution pattern of IRBP immunoreactivity until loss of photoreceptor cells becomes pronounced at later stages of the disease. Many of the remaining cells at this time are probably cone elements although they are structurally changed. Double labeling with IRBP and S antigen demonstrates, in many but not all, the presence of both proteins in the same cell body. Immunocytochemistry clearly demonstrated the presence of IRBP in remaining photoreceptor cells at late stages of the disease. Thus, the biochemically measured loss of IRBP appears to be a complex process neither directly dependent on the loss of photoreceptor outer segments and reduced interphotoreceptor matrix space (e.g. there is a sustained IRBP level in rodless rds mutants) nor simply due to cell death (e.g. in the rd/rd,rds/rds mutant, IRBP loss significantly precedes cell loss). That this IRBP is mainly intracellular, however, may indicate an abnormality in secretion which, combined with other factors, induces a degenerated and less differentiated phenotype. PMID- 3044811 TI - Immunocytochemical localization of laminin, type IV collagen and fibronectin in rat retinal vessels. AB - Laminin, type IV collagen and fibronectin have been identified as major components of the basement membrane (basal lamina) in various tissues. These antigens have also been identified in retinal vessels by light microscopic immunofluorescence but their precise location could not be determined at this level of resolution. In this study, we examined the localization of these constituents at the ultrastructural level using the protein A-immunoperoxidase technique. The basal lamina of all retinal capillaries, arterioles and venules was immunostained after exposure to antisera against laminin, type IV collagen and fibronectin. Staining was localized to the lamina densa, which appeared as a single or double layer. Immunostaining for fibronectin showed the weakest activity. The reaction was also seen in discrete patches between endothelial cells and pericytes. The inner limiting membrane of the retina was also reactive for laminin and type IV collagen but not for fibronectin. The results indicate that laminin, type IV collagen and fibronectin are components of the basal lamina in all types of retinal vessels. The presence of fibronectin at the endothelial pericyte interface suggests that this protein may promote adhesion between cells and thus help to maintain the integrity of the vessel wall. PMID- 3044812 TI - Changes in the major intrinsic polypeptide (MIP26K) during opacification of the Emory mouse lens. AB - Antisera against synthetic peptides corresponding to various regions of the Main Intrinsic Polypeptide (MIP26K) of fiber lens membranes have been used to probe Western blots of Emory mouse lens proteins resolved by SDS-polyacrylamide gel electrophresis. When compared with clear lenses from control animals of approximately the same age, the MIP26K component from Emory mouse lenses demonstrated no quantitative changes in the binding of anti-MIP26K256-263 and anti-MIP26Kwhole sera. In contrast, the MIP26K component from Emory mouse lenses bound significantly better to two other antisera directly against other parts of the molecule (antiMIP26K229-237 and anti-MIP26K252-259). Furthermore, this increase in binding was approximately proportional to the degree of lens opacification. Together, these results demonstrate that during the opacification process of the Emory mouse lens, there occur covalent changes in the MIP26K molecule that, in part, may mimic those occurring in the human senile cataract. PMID- 3044813 TI - Problems and prospects in morphogenesis. PMID- 3044814 TI - A physico-chemical approach to morphogenesis: the roles of inorganic ions and crystals. AB - We consider morphogenesis with special references to the development of mineral frameworks, organic filamentous structures and the location of enzymes, including ion-pumps, in membranes. Starting from a description of the morphology of inorganic crystals we analyse so-called equilibrium growth, i.e. growth at constant shape, both outside and inside biological systems. It is shown that an initial small spherical cell in which linear, ordered, inorganic or organic features are built will become distorted. The distortion is due to stresses which affect membrane curvature and consequently rearrange enzymes in membranes. The cell system can rapidly attain a steady-state of development, ('equilibrium') growth, of fixed morphology. After a considerable growth period the cell may cease to grow or the steady state may be broken and a transition can then occur to a quite new morphology. Examples are taken mostly from unicellular organisms but the ideas apply to multi-cellular systems. PMID- 3044815 TI - Gastrulation in birds: a model system for the study of animal morphogenesis. PMID- 3044816 TI - Physical fields and cellular organisation: field-dependent mechanisms of morphogenesis. PMID- 3044820 TI - 'Auto'-immune neutropenia after allogeneic bone marrow transplantation unresponsive to conventional immunosuppression but resolving promptly after splenectomy. AB - A 32-year-old male presented with isolated neutropenia 6 months after allogeneic bone marrow transplantation for CML from his HLA-matched brother. The presence of granulocyte-specific IgM and IgG antibodies in the patient's serum indicated an immune-mediated basis for the neutropenia. A variety of manoeuvres to suppress antibody production or to reduce peripheral destruction, including high-dose intravenous immunoglobulins 400 mg/kg (total 24 g) on 5 consecutive days, prednisolone 80 mg for 10 d and plasmapheresis on 3 consecutive d, failed to raise the neutrophil count. Splenectomy, however, resulted in a prompt and sustained rise of neutrophils to normal values. PMID- 3044817 TI - Neuronal and glial markers of the central nervous system. AB - This brief review evaluates the expression of cell-specific markers on differentiated neural cells and, where necessary, on their developing precursors. Within these limitations only the commonly used markers are discussed and those deemed unequivocal are only briefly appraised. PMID- 3044819 TI - Immunological phenotype of lymphoid cells in regenerating bone marrow of children after treatment for acute lymphoblastic leukemia. AB - Bone marrow samples from 8 children treated for acute lymphoblastic leukemia (ALL) were investigated at cessation of cytostatic treatment and during 18 months thereafter. The course of the percentage of lymphoid cells and characterization of these cells by means of monoclonal antibodies, peanut agglutinin (PNA) binding and S-phase determination are shown. The percentage of lymphocytes rises in the first 1.5 months, followed by a non-significant decline. The percentage of cells in S-phase is higher at 0 months than at 6, 15 and 18 months. The percentage of T cells does not change significantly. In the first 1.5 months a sudden rise in the percentage of common-ALL-antigen (cALLA)-positive lymphocytes occurs. The number of B-cells rises to a peak at 6 months. PNA positively increases to a maximum at 3 months and is correlated with positivity for markers of the B-cell lineage. The percentages of B-cells, cALLA-positive, and PNA-positive lymphocytes do not change significantly after they reach their maximum values and are still high at 18 months. Our results show that after cessation of chemotherapy for ALL a lymphoid cell regeneration occurs in the bone marrow consisting of cells of the B cell lineage; many of these are cALLA-positive, but are discernible from their malignant counterparts by PNA-positivity. PMID- 3044821 TI - Bone marrow transplantation for myeloblastoma. AB - A 20-yr-old man with bulky mediastinal and retroperitoneal tumour masses identified as myeloblastoma is described. After a partial remission was induced by aggressive chemotherapy, mediastinal irradiation and retroperitoneal tumour resection, the patient received an allogeneic marrow graft from his HLA-identical sister. The conditioning regimen consisted of high-dose busulfan and cyclophosphamide. The patient has a well-controlled secondary chronic graft versus-host disease. He is in unmaintained complete remission and in good general condition at 20 months post-transplantation. PMID- 3044822 TI - [Analysis of the polymorphism of drugs]. PMID- 3044818 TI - The dopaminergic innervation as observed by immunohistochemistry using anti dopamine serum in the rat cerebral cortex. AB - By using an antiserum raised against dopamine bound to bovine serum albumin, thinner dopamine-labeled nerve terminals were visualized immunohistochemically within neocortical areas, in addition to well-documented dopaminergic innervation into the prefrontal and limbic cortices. PMID- 3044823 TI - [Calmodulin: its physiological and pharmacological properties]. PMID- 3044824 TI - [Pharmacological agents that interrupt early-stage pregnancy]. PMID- 3044825 TI - The molecular cloning of the chromogranin A-like precursor of beta-granin and pancreastatin from the endocrine pancreas. AB - The cDNA encoding the precursor form of the chromogranin A-related proteins, beta granin and pancreastatin, was obtained by immune screening of rat insulinoma and pancreatic islet cDNA libraries. The sequence was virtually identical to that of rat adrenal chromogranin A, suggesting that the different molecular forms of chromogranin A immunoreactivity found in adrenal medulla and endocrine pancreas are related to differences in post-translational proteolytic processing. The rat chromogranin A, unlike its bovine and human counterparts, contained a 20-residue glutamine sequence inserted within the N-terminal beta-granin sequence. Although the encoding CA(G/A) repeat recurs frequently in the rat genome, the rat chromogranin A molecule appears to be the product of a single gene and mRNA transcript. PMID- 3044826 TI - Structure-function relationship in Escherichia coli translational initiation factors. Characterization of IF1 by high-resolution 1H-NMR spectroscopy. AB - Escherichia coli translational initiation factor IF1 was studied by 1H-NMR spectroscopy at 400 MHz. IF1 displays a very well resolved spectrum in both aromatic and aliphatic regions. Other spectral characteristics include relatively narrow resonance lines and lack of relevant cross-relaxation phenomena. The resonances of the aromatic residues, in particular of the two His and two Tyr, were assigned by selective chemical modifications and spectroscopic techniques to individual residues in the protein sequence. The relative mobility of various residues of IF1 has been evaluated on the basis of the spin-lattice relaxation times which are rather short and homogeneous. Overall the factor appears to have a complex secondary and tertiary structure and to be a flexible protein whose residues have a high degree of internal mobility. PMID- 3044827 TI - Circular dichroism and fluorescence of polyethylene glycol-subtilisin in organic solvents. AB - Subtilisin Carlsberg has been made soluble in organic solvents such as dioxane and acetonitrile by covalent linking to polyethylene glycol. Far-ultraviolet circular dichroism and intrinsic protein fluorescence have shown that subtilisin dissolved in dioxane, in which the enzyme is active and highly stable, maintains the native secondary structure as well as the native microenvironment for tyrosyl residues. In acetonitrile subtilisin undergoes conformational changes that cause enzyme inactivation and precipitation. PMID- 3044828 TI - Reconstitution of the active rat liver 60 S ribosomal subunit from different preparations of core particles and split proteins. AB - Proteins extracted from the 60 S rat liver ribosomal subunit with 50% ethanol/0.5 M K Cl produced only a partial reactivation of the corresponding core particles. In contrast, the same split proteins were able to reactivate the core particles prepared with dimethyl-maleic anhydride (DMMA) to the same level as that observed using the DMMA-split proteins, i.e. 60-80% of the control according to the catalytic activities tested. Comparative analysis of the two split protein fractions showed only four common proteins: P1-P2, which alone restored part of the activities, especially the EF-2-dependent GTPase one, and L10a, L12, which must be responsible for the additional reactivation. The poor ability of the ethanol/KCl core particles to be reactivated was shown to be probably related to a conformational alteration which destabilized the 5 S RNA-protein complex. Proteins present in the ethanol/KCl wash of Saccharomyces cerevisiae 60 S subunits were found to be partly active in subunit reconstitution using rat liver DMMA core particles. PMID- 3044829 TI - Effects of glucose on insulin release and 86Rb permeability in cultured neonatal and adult rat islets. AB - Glucose-induced insulin release and modifications in 86Rb outflow were studied in cultured neonatal and adult rat islets. The dose-response curve for neonatal islets was steeper than for adult islets and the maximal response was clearly shifted towards lower glucose concentrations. In neonatal islets, glucose-induced insulin release was inhibited by the Ca2+-channel blocker, nifedipine. In the absence of glucose, the 86Rb outflow from neonatal islets was lower than from adult islets. Also, the glucose-induced reduction in 86Rb outflow was less pronounced in neonatal islets. Altered K+ permeability in the B-cell membrane could explain the change in glucose sensitivity of neonatal islets. PMID- 3044830 TI - Synthesis and biological characterization of monocyte-derived neutrophil chemotactic factor. AB - MDNCF is a human monocyte-derived, 72-residue chemotactic peptide, which has sequence similarity with members of a family of pro-inflammatory cytokines. The peptide was synthesized by the solid-phase method, and is identical to the natural peptide in amino acid composition, sequence and chemotactic potency. MDNCF forms two loops via a neighboring pair of disulfide bridges, the probable locations of which are residues 7-34 and 9-50. Reduction and alkylation eliminated chemotactic activity. MDNCF fragments 7-37, 30-72 and 17-72 were all biologically inactive. The data suggest that the region of the clustered pair of disulfide bridges is important for biological activity. PMID- 3044831 TI - Two rat homologues of human cystatin C. AB - Two immunochemically related forms of cystatin C-like inhibitors which differ in their Mr app and isoelectric point have been found both in urine and seminal vesicles of rats. Amino-terminal sequences of these two cystatins are identical within the same fluid and exhibit a high degree of homology with that of human cystatin C. However, cystatins C purified from urine lack eight residues at their amino-terminal end when compared to those of seminal vesicles. The occurrence of two cystatin C-like components in rat fluids has been found to be due to the presence of a glycosylated form reported here as cystatin Cg which specifically binds concanavalin A and is susceptible to endo-beta-N-acetylglucosaminidase treatment. PMID- 3044832 TI - Rectal endosonography, a new technique for the preoperative staging of rectal carcinoma. AB - Endosonography of the rectum is a new and exciting technique which is having an increasing impact on the surgical and radiological community and is likely to have two principal applications in patients with rectal cancer. Firstly in the preoperative staging of the disease it has been shown to provide an objective reproducible method of assessment allowing the more accurate planning of treatment whether purely surgical or including some form of adjuvant treatment. Secondly in the follow up of patients who have already had surgery or radiotherapy it may be possible to detect in particular extrarectal local reoccurence at an early and treatable stage. Initial results with this technique are extremely promising but further long term studies are now needed to see whether it will improve overall survival or decrease local recurrence. PMID- 3044834 TI - Prophylactic hyperthermic limb perfusion in stage I melanoma. AB - Ninety-three patients with stage I primary cutaneous malignant melanoma of the lower limb were treated by wide local excision and hyperthermic isolated regional perfusion with melphalan (L-phenylalanine dihydrochloride) in a prospective non randomized study between 1976 and 1982. Eighteen patients (19.4%) developed recurrent melanoma. Nine had recurrent regional disease, one with in transit metastases and eight with positive regional nodes. Nine patients developed distant metastases. No patient had locally recurrent disease. This series confirmed the close correlation between tumour microstaging, melanoma recurrence and survival. Seventy-nine per cent of patients were disease-free at 5 years. Males had deeper lesions (mean 4.56 mm) and increased recurrence (33%) than females (mean 3.36 mm and 13%). Superficial spreading melanoma had the most favourable prognosis of the three histological types. Overall survival was 83% (female 86%; males 64%) at 5 years. Significant morbidity occurred in two patients with deep vein thrombosis. Adjuvant therapy using hyperthermic regional perfusion provides improved local and intransit control of limb melanoma. PMID- 3044833 TI - Polyadenylic-polyuridylic acid as adjuvant in the treatment of operable breast cancer: recent results. AB - A randomized trial of polyadenylic-polyuridylic acid (Poly(A).Poly(U) given as an adjuvant in the treatment of operable breast cancer, has included 300 patients of the Institut Gustave-Roussy from September 1972 to December 1979; 145 patients were allocated to conventional treatment alone and 155 to conventional treatment plus Poly(A).Poly(U). Reviews after mean periods of follow-up of 50 and 87 months were previously published. The present review performed after a mean follow-up period of 111 months confirmed a significant increase in the overall survival of patients with invaded nodes treated with Poly(A).Poly(U). The best results were achieved in the subset of patients with up to three affected nodes who showed a significant increase of both overall and relapse-free survival. The benefit seemed to be greater in postmenopausal women (P = 0.07). Present status of other ongoing trials of adjuvant Poly(A).Poly(U) is presented. PMID- 3044836 TI - Laser Doppler flux reappearance time (FRT) in patients with lower limb atherosclerosis and healthy controls. AB - Laser doppler flowmetry was used to examine the skin circulation in the lower limb during postischaemic reactive hyperaemia. Flux reappearance time (FRT), the time from tourniquet deflation to the start of the hyperaemic response, was determined from the recorded curves, and the aim of the study was to investigate whether FRT is dependent on total limb vascular resistance or to the resistance in a segment of the limb. FRT was first compared with the clinical situation of the examined subjects, and with the resting ankle blood pressure index (API). Controls had an immediate hyperaemic response (FRT less than or equal to 3 s), while patients with critical ischaemia had prolonged FRT (greater than 48 s). Among the claudicators there was no significant correlation between API or ankle blood pressure and FRT. When FRT was compared with angiography, all claudicators who only had significant atherosclerosis proximal to the tourniquet, had FRT values within the control range. Patients who only had distal atherosclerosis had prolonged values (greater than 15 s), indicating that FRT is independent of proximal atherosclerosis and reflects the vascular resistance in the arteries in the segment between the tourniquet and the measuring probe. This interpretation was supported by the reduction of pathologically prolonged FRT when the distance between the tourniquet and the measuring site was reduced. We conclude that FRT as measured by this technique seems to reflect the vascular resistance in the run off arteries distal to the tourniquet. PMID- 3044835 TI - Francisco Gentil Martins (1927-1988). PMID- 3044837 TI - The reliability of duplex derived haemodynamic measurements in the assessment of femoro-distal grafts. AB - The aim of this study was to assess the reliability of flow and velocity measurements obtained in the postoperative period by duplex scan monitoring of femoro-distal bypass grafts. Three points along each of 24 grafts were studied by 2 independent observers using a Diasonics 300 DRF duplex scanner. Results obtained for diameter, peak systolic velocity (PSV) and flow were compared. The mean PSV for all points studied was 70 cm/s (S.D. = 14.5): comparing observers the calculated experimental error (EE) for all points was 11 cm/s (coefficient of EE = 15%). In contrast, the experimental error for flow was 44.5 ml/min (coefficient of EE = 31). The mean flow for all grafts was 142 ml/min (S.D. = 66.5). Flow at the different points of individual grafts could also be compared since routine postoperative intravenous angiography showed all grafts to be devoid of branches. The coefficient of experimental error for flow was 58%. Four of the grafts studied were found to have asymptomatic stenoses. These 4 grafts had low flows (mean = 42 ml/min) but so did 7 other non-stenotic grafts. Although PSV measurements entailed a 15% error, the mean change in PSV in relation to the stenoses was 350%. These data suggest that flow is an unreliable measurement. PSV is more reliable, changes exponentially with luminal diameter and can detect, and perhaps grade, asymptomatic stenoses. PMID- 3044839 TI - [Yersinia infection]. PMID- 3044838 TI - [40th anniversary of the World Health Organization]. PMID- 3044840 TI - [Prof. Boris Vladimirovich Verkhovskii (on the 125th anniversary of his birth)]. PMID- 3044841 TI - Alterations in carbohydrate metabolism as they apply to reproductive endocrinology. AB - This review has characterized the current state of knowledge of four clinical situations in which an interrelationship of gynecology, endocrinology and carbohydrate metabolism is recognized. The literature contains conflicting descriptions of changes in glucose homeostasis during the menstrual cycle and while using birth control pills. Physiologic changes in receptor number have been demonstrated in each of these situations, so failure to observe differences using glucose tolerance testing may reflect an in vivo homeostatic response to changes in these hormone levels. Thus, in vivo identification of alterations in carbohydrate metabolism induced by endogenous or exogenous steroids may require utilization of models that prevent these homeostatic mechanisms. The association between hyperandrogenism and hyperinsulinism has been better characterized, but the relationship is complicated by the frequent coexistence of obesity. The association may be due to insulin-stimulated ovarian androgen production, and insulin insensitivity may reflect a postreceptor defect. Insulin and its metabolic effects have also been implicated in ovulatory dysfunction in women with diabetes mellitus and identified as a factor affecting all levels of the hypothalamic-pituitary-ovarian axis. A clearer understanding of these relationships and their application to clinical management await further study. PMID- 3044842 TI - Transvaginal ultrasonography for the diagnosis of ectopic pregnancy. AB - The transvaginal (TVU) and transabdominal (TAU) ultrasound approaches were compared in their ability to identify by direct visualization the adnexal mass of ectopic pregnancy. There were 22 patients who had a surgically proven ectopic pregnancy. The TAU approach identified the adnexal mass in 50% and the TVU approach in 91% of the patients (P less than 0.01). Below both previously reported threshold titers for the expected TAU intrauterine sac visualization, at 6500 mIU and 3600 mIU, respectively, the TVU approach allowed the identification of significantly more ectopic adnexal masses than the TAU approach. The results of the present study demonstrate the increased efficacy of TVU over TAU in the direct identification of the adnexal mass associated with ectopic gestation. PMID- 3044843 TI - Follicular aspiration and in vitro fertilization associated with pelvic reconstructive surgery. AB - Between August 1982 and May 1987, 103 patients underwent in vitro fertilization embryo transfer (IVF-ET) in association with pelvic reconstructive surgery for infertility. Follicular stimulation was induced with clomiphene citrate and laparotomy scheduled day 12 to 15 of the menstrual cycle. Ultrasound measurements of follicular diameter and number of follicles were obtained on the day of human chorionic gonadotropin (hCG) administration, and laparotomy and ovum retrieval performed 36 hours later. Embryo transfer was performed 48 to 72 hours after insemination. Patients were treated postoperatively with intramuscular progesterone. In addition to evaluating the overall pregnancy rate, the outcome of patients having one or more follicles greater than or equal to 1.4 cm in mean diameter (group A) were compared to those in group B (no follicles greater than or equal to 1.4 cm in diameter). The number of oocytes obtained and the fertilization rate and polyspermic fertilization rate were not significantly different between groups; 10.1% of patients in group A conceived but no patient conceived in group B, yielding an overall pregnancy rate of 8.7%. These data suggest that physicians having IVF-ET at their disposal offer patients IVF during pelvic reconstructive surgery. PMID- 3044844 TI - Doyne Lecture: Current concepts in orbital disease. PMID- 3044845 TI - Adjustable sutures. PMID- 3044846 TI - Faden operation (posterior fixation sutures). PMID- 3044847 TI - Epikeratophakia: clinical results and experimental development. AB - The clinical course and visual, refractive, and keratometric results of a consecutive series of epikeratophakia procedures carried out by the author are presented. Indications for the procedure included keratoconus and adult and paediatric aphakia. Follow up time ranged from two to fourteen months. The first five patients operated on received commercially obtained cryolathed lenticules. The final three cases received lenticules which were lathed by the author at room temperature using a recently developed technique. PMID- 3044849 TI - Ultrasonic Doppler studies of the vitreous. AB - Simultaneous 'real-time' B-mode and doppler ultrasound techniques were used to study movements of the detached vitreous gel which occurred during duction movements of the eye. The velocity of sound scatterers within the vitreous gel were measured and it was possible to differentiate quantitatively gels which were 'stiff' from those which were 'sloppy'. Information obtained using this technique may throw light on the pathogenesis of rhegmatogenous retinal detachment and assist in the quantitative assessment of retinal mobility in eyes with vitreoretinopathies. PMID- 3044848 TI - Emergency presentation of corneal graft patients. AB - One hundred and forty-two patients with corneal grafts made 205 visits to the Accident and Emergency Department of Moorfields Eye Hospital during a twelve month period. One-third of cases showed a graft rejection episode while another third had a problem directly related to the graft. The figures emphasise the need for continuing patient education so that graft problems may be dealt with promptly. PMID- 3044850 TI - Doppler ultrasound studies of the ophthalmic artery. AB - The doppler frequency shift of ultrasound pulses scattered off red blood cells in the ophthalmic artery can be detected and used as an index of velocity of flow in the artery. The doppler shift is shown to be responsive to changes in ocular blood flow induced by changes in mean arterial blood pressure at the level of the eye and changes in intraocular pressure. The technique may be useful in the study of eye disease in which blood flow is altered. Doppler frequency shifted signals have also been detected within the coats of the eye. PMID- 3044851 TI - Ocular autonomic nerve function in proliferative diabetic retinopathy. AB - Ocular autonomic nerve function was assessed in 28 diabetic patients with proliferative retinopathy (PDR) and 61 age- and sex-matched control subjects, by measurement of the pupil cycle time and determination of autonomic denervation hypersensitivity of the iris. Sustained pupil cycle time was absent in 88.5 per cent of PDR compared with 9.8 per cent of control subjects (p less than 0.001). Pupil constriction in response to 2.5 per cent methacholine--indicative of parasympathetic denervation hypersensitivity--was significantly increased in PDR (p less than 0.001), whilst pupil dilation in response to 0.5 per cent phenylephrine--indicative of sympathetic denervation hypersensitivity--was also significantly higher in PDR (p less than 0.001). Pupil reflexes were abnormal in 88.5 per cent of diabetics with proliferative retinopathy, with established autonomic denervation hypersensitivity in 57 per cent of patients. PMID- 3044852 TI - [Development of a highly sensitive assay for serum thyroglobulin using avidin biotin system and its clinical application]. AB - The authors have developed a highly sensitive sandwich-type enzyme immunoassay (EIA) for serum thyroglobulin using the biotin-avidin system. The sensitivity of this EIA was 0.1 ng/ml. The intra- and inter-assay coefficient of variation was 4 15% and 4-11%, respectively. Human Tg in serum was detectable in 100% of 63 normal subjects, and the normal range was from 2.3 ng/ml to 47.4 ng/ml. The recovery rate was very good. Then we applied this EIA to the follow-up study of patients with thyroid cancer. Serum Tg levels were serially determined in 15 patients with thyroid cancer who had undergone thyroidectomy (n = 9 subtotal, n = 6 total) and were receiving thyroid hormone suppression therapy. It was found that there were three groups in regard to the relationship between serum TSH and Tg. The first group showed normal TSH and normal Tg levels (n = 4). The second group showed TSH levels that were lower than the lower limit of the assay, whereas Tg levels were within the normal limit (n = 5). The last group showed both TSH and Tg that were lower than the lower limit (n = 6). In one case in the last group, recurrence of thyroid cancer was forecast by the change of Tg levels (from below 0.1 ng/ml to 5.9 ng/ml). Therefore, it is important to suppress serum Tg and TSH levels as much as possible in order to obtain the best usefulness of serum Tg determination in the follow-up study of thyroid cancer in patients who have undergone subtotal thyroidectomy or have residual thyroid tissue. We also measured Tg levels in patients with untreated Graves' disease. Twelve patients were negative for anti-Tg antibody and 18 were positive for anti-Tg antibody. Even in the antibody positive cases, serum Tg could be detected and their levels could be calculated from the recovery studies. The mean serum Tg levels in antibody positive cases were lower than in negative cases. These studies showed that our sensitive EIA for serum Tg determination was useful in practice in patients with thyroid disease. PMID- 3044853 TI - Recurrent perianal streptococcal cellulitis. PMID- 3044855 TI - [Backward and analysis. Partial denture scrutinized intently]. PMID- 3044854 TI - Venous reconstruction in venous insufficiency syndrome. PMID- 3044856 TI - [Gysi Prize 1978. Work of a prize winner]. PMID- 3044857 TI - [Esthetics and precision? Margins of ceramic crowns]. PMID- 3044858 TI - [3. Biomechanical waxing technique. Practice after the pattern of nature. Twelve further work steps]. PMID- 3044859 TI - [Precision casting technic for fixed partial denture. From wax model to firing investment]. PMID- 3044860 TI - [Direct cast attachment. Precision with simple materials]. PMID- 3044862 TI - [Precision casting technic for fixed partial dentures. Preheating--pouring- deflasking--cleaning]. PMID- 3044863 TI - Free insulin profiles during intensive treatment with biosynthetic human insulin. AB - 24 h profiles of free insulin and glucose were determined in insulin-dependent diabetic patients on intensive insulin regimens with biosynthetic human insulin, either as continuous subcutaneous insulin infusion (CSII) with mealtime bolus doses (n = 6), or intensified conventional insulin therapy (ICIT) with preprandial injections of regular insulin and intermediate-acting insulin at bedtime (n = 6). The free insulin profiles were similar to the normal profiles but there were some important differences: CSII gave hyperinsulinaemia at daytime compared to normal people (p less than 0.05) and also to ICIT (p less than 0.005); ICIT but not CSII gave hyperinsulinaemia at midnight (p less than 0.05) whereas fasting free insulin was too-low to keep blood glucose normal; the insulin peaks after the bolus doses were retarded with a maximum after 30-90 min and a return to the basal level after 5-7 h (ICIT) or 8-9 h (CSII). The height of the insulin peaks were of similar magnitude at all meals and did not differ significantly between ICIT, CSII, and normal people. Time-to-peak was dependent on the injection level. We conclude, that intensive regimens with biosynthetic hum insulin do not give normoinsulinaemia but insulin profiles that resemble physiology. Biosynthetic human NPH insulin may be rather short-acting for overnight glucose control. The interval that should be recommended between preprandial insulin dose and meal may vary depending on the preinjection insulin level. PMID- 3044861 TI - [Crozat method of late orthodontic treatment]. PMID- 3044864 TI - Carbohydrate metabolism and plasma levels of insulin and glucagon in patients with atherosclerotic vascular disease. AB - In non-obese, non-diabetic patients suffering acute myocardial infarction, angina pectoris, previous myocardial infarction and peripheral vascular disease, the plasma levels of glucose, insulin, C-peptide and glucagon were determined in basal condition and during an intravenous glucose tolerance test. In the four groups there was a high frequency of glucose intolerance. Basal hyperinsulinism was present in all groups; in groups; in those which maintained normal glucose tolerance there was a high B-cell response to the sugar. Basal hyperglucagonemia was found in the early stage of acute ischemic heart disease, in patients with previous myocardial infarction and in those with peripheral vascular disease. The elevated plasma glucagon levels may play a role in the complex disturbance of carbohydrate metabolism present in patients with atherosclerotic vascular disease. PMID- 3044866 TI - [Diabetogenic tropical pancreatitis]. AB - The tropical calcifying pancreatitis and/or fibrous pancreatitis are responsible for a number of cases of juvenile insulin-dependent diabetes in the Third World countries. World wide distributed in the tropical areas of Asia, Africa and South America, they can also be observed in Europe, in migrants from these countries. Intensive epidemiological and biochemical studies are currently developed in order to shed light on the many obscure points. Classification of the typical calcifying pancreatitis and the related syndromes is a matter of debate. The pathological basis is calcification of the pancreas and echography of the gland may become a cheap convenient relatively specific tool for epidemiology. The clinical syndrome consists of chronic painful pancreatic episodes since childhood, associated with pancreatic exocrine insufficiency, followed by the onset, during adolescence, of diabetes mellitus, which is most of the times insulin dependent. Patients' history is free of chronic alcoholism, but includes constantly chronic caloric and proteic malnutrition. Although insulin dependent this diabetes in not prone to ketosis, due presumably to carnitine deficiency and relative glucagon deficiency (or suppressibility). Insulin resistance is traditionally noted, the pathophysiology of which is unknown. The mechanism of calcification appearance is also undetermined. Either a deficiency in pancreatic stone protein, or the toxic effect of cyanogen glucosides present in cassava and other tropical foodstuffs, or the malnutrition-related deficiency in sulphur containing aminoacids may be causal factors. No valid experimental model of the disease is available. PMID- 3044865 TI - Insulin sensitivity and metabolic clearance rate of insulin in familial multiple lipomatosis. AB - Intolerance to glucose in certain kinds of lipomatosis is well documented. This article describes a euglucaemic hyperinsulinaemic clamp study of alterations in glucose and/or insulin metabolism in four members of a single family with familial multiple lipomatosis. Fifteen normal subjects were studied as controls. The four patients exhibited no alteration in tolerance to orally administered glucose. When a Biostator Glucose-Controlled Insulin Infusion System (GCIIS) was used to clamp glycaemia at 4.44 mmol/L with successive insulin infusion rates of (a) 0.5 (b) 1.0 or (c) 5.0 mU/kg/min, there was no difference between patients and controls as regards the value of M, the rate of glucose infusion, but the concentrations of immunoreactive insulin recorded during the last 40 minutes of each phase of the clamp were greater in patients than in controls (45 +/- 2 vs 27 +/- 2 uU/mL (p less than 0.01), 83 +/- 2 vs 60 +/- 5 uU/mL (p less than 0.05) and 537 +/- 48 vs 377 +/- 25 uU/mL (p less than 0.05) for insulin infusion rates (a), (b) and (c) respectively), and the ratio M/IRI was consequently smaller for patients than controls (1.92 +/- 0.41 vs 3.06 +/- 0.19 (p less than 0.05) for an insulin infusion rate of 5 mU/kg/min). The metabolic clearance rate of insulin was likewise slower in patients than controls (p less than 0.01). It is concluded that the four patients studied (all members of the same family) have sub-normal sensitivity to insulin secondary to a sub-normal metabolic clearance rate for insulin. PMID- 3044867 TI - Mixtures of soluble insulin with zinc-suspension or protamine insulin have different effect on blood glucose and free insulin levels in insulin dependent diabetes. PMID- 3044868 TI - Analgesic effect of aspirin, mefenamic acid and their combination in post operative oral surgery pain. AB - A double-blind randomized single dose study of the analgesic effects of 650 mg aspirin, 250 mg mefenamic acid, the combination of 650 mg aspirin and 250 mg mefenamic acid and placebo on 120 patients with pain following oral surgery was conducted. Patients evaluated their pain intensity and extent of pain relief at 1, 2, 3 and 4 h after drug administration. For most parameters, including the sum of the pain intensity differences and the sum of the hourly pain relief scores, each of the drugs was more effective than placebo. Aspirin-mefenamic acid in combination was more effective than both drugs alone, and aspirin and mefenamic acid alone were equally effective for most of the analgesic variables. PMID- 3044869 TI - Effect of ibuprofen on blood pressure control by propranolol and bendrofluazide. AB - The effect of 1600 mg/day ibuprofen in two groups of patients with hypertension controlled by either propranolol or bendrofluazide was studied in a double-blind, double-placebo, randomized crossover trial. No significant difference in blood pressure was found at the end of the crossover period in either group, suggesting that the routine co-administration of ibuprofen does not attenuate the anti hypertensive effect of thiazide diuretics or propranolol. Significant weight gain, attributable to fluid retention, had occurred in the bendrofluazide treatment group by the end of the drug-free washout period. No significant change in mean weight occurred in the crossover stages in either group, although substantial weight gain was noted during ibuprofen treatment in two patients given bendrofluazide and one given propranolol. Biochemical variables were unaffected by ibuprofen throughout the crossover period. This study suggests that ibuprofen may be administered routinely to patients receiving thiazides or propranolol without loss of control of the anti-hypertensive action of these drugs but it is recommended that individuals are monitored for possible weight gain or an increase in diastolic blood pressure. PMID- 3044871 TI - A double-blind clinical comparison of two dosage schedules of cefatrizine in children. AB - A double-blind, randomized clinical trial was carried out to compare the effectiveness of twice daily versus once daily administration of the cephalosporin, cefatrizine, in paediatric outpatients with bacterial infection of the respiratory tract. Thirty children were studied, aged 7 years 2 months (range, 4-12 years). They were given 75 mg/kg.day cefatrizine either once daily or twice daily at 12 h intervals for 8 days. Fever, clinical symptoms, bacterial eradication and overall tolerance were evaluated. No significant differences were observed between once daily or twice daily administration. This is in agreement with other studies carried out on adults. It is concluded that cefatrizine may be given to paediatric out-patients for the treatment of bacterial infection of the respiratory tract only once daily with good clinical and overall results. PMID- 3044872 TI - Clinical efficacy of codergocrine mesylate in children with learning difficulties. AB - A double-blind placebo-controlled study in children showed codergocrine mesylate to be effective in improving cognitive functions and behavioural symptoms associated with learning disorders. Forty randomly grouped children of either sex were given an increasing dosage of codergocrine mesylate and followed up for 12 weeks. A significant improvement was noted in speech (acquisition of new words, comprehensibility/meaningfulness of speech), sociability, attention/concentration, comprehension and memory. Improvement in behaviour (emotional lability and cooperativeness) was also noted. Problems of assessing cognitive progress in very young children with culturally appropriate methods were encountered. PMID- 3044870 TI - Night pain and morning stiffness in osteoarthritis: a crossover study of flurbiprofen and diclofenac sodium. AB - Forty patients took part in an observer-blind multiple dose crossover study to compare the efficacy and tolerability of 300 mg flurbiprofen (100 mg twice a day orally and 100 mg suppository at night) with 75 mg diclofenac sodium (25 mg twice daily orally and 25 mg suppository at night) in the relief of night pain and morning stiffness in patients with osteoarthritis. Equal numbers of patients were randomized to receive one of the treatments for the first 7-day treatment period and then the other treatment for the second 7-day treatment period. There were significant differences in favour of flurbiprofen for the reduction in night pain, improvement in quality of sleep and patients' assessment of overall improvement on treatment. Two patients withdrew from the study due to side effects experienced whilst taking diclofenac sodium during the first treatment period. Eight patients in all reported a total of 18 side-effects during this study. Six patients reported 12 side-effects whilst taking diclofenac sodium compared with three patients who reported six side-effects whilst taking flurbiprofen. This study confirms the good efficacy and tolerability of flurbiprofen in the symptomatic relief of osteoarthritis. PMID- 3044873 TI - A clinical evaluation of clavulanate-potentiated amoxycillin in the management of moderate to severe respiratory tract infections. AB - This study investigated the effects of clavulanate-potentiated amoxycillin in the management of moderate to severe respiratory tract infections. A total of 34 patients aged 10-70 years were studied to establish the clinical and bacterial efficacy of treatment and screened for unwanted effects and abnormal laboratory findings. Clavulanate-potentiated amoxycillin was given in dosages of 375 or 750 mg twice daily, orally, for a mean of 13 days (range 3-30 days). Patients were assessed at the end of 2 weeks and again on discharge from hospital (if later) for up to 6 months. Overall diminution in the number of symptoms was recorded at the end of the study; 78% of symptoms had disappeared after 2 weeks. The treatment proved highly effective in controlling respiratory tract infections in 94% of patients. The necessity for multiple drug therapy and parenteral administration was also avoided. PMID- 3044874 TI - Dipyrone versus paracetamol: a double-blind study in typhoid fever. AB - In a double-blind study in patients with typhoid fever 25 patients received 500 mg dipyrone and 28 received 500 mg paracetamol. Rectal temperature and pulse records were monitored every 30 min. The onset of anti-pyresis in patients given dipyrone (30 min) was significantly different from those given paracetamol (1 h). The area under the time-temperature curves was significantly greater for patients given dipyrone. The sum of the reduction in temperature at all times significantly favoured patients who had been given dipyrone. Both treatments were well tolerated. PMID- 3044875 TI - Clinical study of recombinant hepatitis B vaccine. AB - The efficacy and safety of a recombinant yeast-derived hepatitis B vaccine were evaluated in 209 subjects after three administrations at 0, 4 and 20 weeks. Subjects were divided into four groups given 5 micrograms vaccine subcutaneously, 10 micrograms subcutaneously, 10 micrograms intramuscularly and 20 micrograms subcutaneously to define the effective dose and to compare the effect of administration. Seroconversion of the antibody to hepatitis B surface antigen after the third vaccination reached 96.6% in the group given 5 micrograms vaccine subcutaneously and 100% in the other groups. The final geometric mean antibody titres were 700 IU/l in subjects given 5 micrograms subcutaneously, 2004 IU/l in those given 10 micrograms subcutaneously, 4674 IU/l in those given 10 micrograms intramuscularly and 3342 IU/l in those given 20 micrograms subcutaneously. In the groups given 10 micrograms, the early seroconversion rate of the antibody to hepatitis B surface antigen and the geometric mean antibody titres after the third vaccination were significantly higher in subjects administered intramuscularly than subcutaneously (P less than 0.05). No major adverse effects were observed and minor reactions were the same as, or less than, those reported for the plasma-derived vaccine. Before and after administration, no significant fluctuation in the yeast antibody titre was observed. These results demonstrate the efficacy and safety of the yeast-derived vaccine, and show that 10 micrograms was the effective dose. PMID- 3044876 TI - Comparison of flavoxate hydrochloride in daily dosages of 600 versus 1200 mg for the treatment of urgency and urge incontinence. AB - Flavoxate hydrochloride at a daily dosage of 600 mg was compared to a daily dosage of 1200 mg for the treatment of unstable bladder. Twenty-seven patients were treated for 4 weeks in a double-blind, randomized, parallel-group trial. Clinically, both schedules were equally successful. In urodynamic terms, however, particularly with respect to uninhibited detrusor contractions, 1200 mg/day was significantly superior to 600 mg/day. Tolerability was excellent for both regimens. The side-effect free treatment of urgency and urge incontinence is of paramount importance for a patient's quality of life. PMID- 3044877 TI - Monoclonal antibodies specific for thiophosphorylated proteins recognize Xenopus MPF. AB - Maturation promoting factor, (MPF), is a crucial regulatory component of the eukaryotic cell cycle. Though it is ubiquitous, MPF has been difficult to purify to homogeneity, and little is known about its physical properties or composition. In an attempt to further characterize and purify this protein, we have isolated five monoclonal antibodies that immunoadsorb MPF activity, and inhibit the activity in solution. However, all the antibodies recognize many proteins in partially purified MPF. We have shown that antibody binding is dependent on previous exposure of the preparation to ATP gamma S. This suggests that the antibodies specifically recognize thiophosphoproteins, although not all thiophosphorylated proteins in MPF are immunoprecipitated. Using one antibody, MPF was partially purified by immunoadsorption chromatography. These experiments provide the first evidence that MPF from Xenopus is a phosphoprotein that becomes thiophosphorylated upon addition of ATP gamma S. PMID- 3044879 TI - A proteinase associated with cortices of rainbow trout eggs and involved in fertilization-induced depolymerization of polysialoglycoproteins. AB - A novel proteinase that acts on polysialoglycoprotein (PSGP) was found in the cortex fraction of the unfertilized eggs of rainbow trout. This enzyme (designated PSGPase) is responsible for specific depolymerization of cortical vesicular 200- to 9-kDa PSGP in vivo upon fertilization. We have succeeded in measuring the enzyme activity in an in vitro system by using 3H-labeled PSGP as the substrate. In the in vitro system PSGPase is active only at concentrations of NaCl below 40 mM and at low temperature (optimum temperature, about 16 degrees C), which are the conditions most suitable for egg activation. PMID- 3044878 TI - Alterations in the Xenopus retinotectal projection by antibodies to Xenopus N CAM. AB - The patterned neural projection from the eye to the optic tectum of lower vertebrates (the retinotectal projection) has been proposed to be ordered by interactions between the optic nerve fibers and their surrounding tissues. To investigate the role of one such defined cell interaction, agarose implants containing antibodies to the neural cell adhesion molecule, N-CAM, were inserted into the tectum of the African clawed frog, Xenopus laevis. Both monoclonal and polyclonal antibodies against N-CAM reversibly and specifically distorted the pattern of the retinotectal projection, decreasing the precision of the projection as determined by electrophysiological techniques as well as decreasing the density of retinal innervation of the tectum and the branching of single axons as determined by horseradish peroxidase tracing. The anatomical effects became maximal at 4 to 6 days after implantation and returned to undetectable levels by 2 weeks, whereas the physiological effects became maximal by 8 to 10 days and a normal physiological map was reestablished within 4 weeks. The results are consistent with the hypothesis that anti-N-CAM antibodies perturb the ongoing growth and retraction of the terminal arbors of the optic nerve fibers, such that a region of the tectum becomes largely denuded of fibers. The physiological defects may then be a consequence both of the initial retraction of optic nerve terminals and of the rapid ingrowth of the perturbed and neighboring optic nerve fibers into the denuded region after the antibodies were cleared from the tectum. These results support the concept of a major role for N-CAM-mediated adhesion during map regeneration and maintenance. PMID- 3044880 TI - Unusual organization of desmin intermediate filaments in muscular dysgenesis and TTX-treated myotubes. AB - Cytoskeletal intermediate filaments were studied in muscular dysgenesis (mdg) and tetrodotoxin-treated inactive mouse embryo muscle cultures during myofibrillogenesis. Both muscular dysgenesis and tetrodotoxin-treated muscles are characterized in vitro by a total lack of contractile activity and an abnormal development of myofibrils. We studied the organization of the microtubule and intermediate filament networks with immunofluorescence, using anti-tubulin, anti vimentin, and anti-desmin antibodies during normal and mdg/mdg myogenesis in vitro. Mdg/mdg myotubes present a heterogeneous microtubule network with scattered areas of decreased microtubule density. At the myoblast stage, cells expressed both vimentin and desmin. After fusion only desmin expression is revealed. In mutant myotubes the desmin network remains in a diffuse position and does not reorganize itself transversely, as it does during normal myogenesis. The absence of a mature organization of the desmin network in mdg/mdg myotubes is accompanied by a lack of organization of myofibrils. The role of muscle activity in the organization of myofibrils and desmin filaments was tested in two ways: (i) mdg/mdg myotubes were rendered active by coculturing with normal spinal cord cells, and (ii) normal myotubes were treated with tetrodotoxin (TTX) to suppress contractions. Mdg/mdg innervated myotubes showed cross-striated myofibrils, whereas desmin filaments remained diffuse. TTX-treated myotubes possessed disorganized myofibrils and a very unusual pattern of distribution of desmin: intensively stained desmin aggregates were superimposed upon the diffuse network. We conclude, on the basis of these results, that myofibrillar organization does not directly involve intermediate filaments but does need contractile activity. PMID- 3044881 TI - The fate of Meckel's cartilage chondrocytes in ocular culture. AB - Modulation of the chondrocyte phenotype was observed in an organ culture system using Meckel's cartilage. First branchial arch cartilage was dissected from fetal rats of 16- and 17-day gestation. Perichondrium was mechanically removed, cartilage was split at the rostral process, and each half was grafted into the anterior chamber of an adult rat eye. The observed pattern of development in nonirradiated specimens was the following: hypertrophy of the rostral process and endochondral-type ossification, fibrous atrophy in the midsection, and mineralization of the malleus and incus. A change in matrix composition of the implanted cartilage was demonstrated with immunofluorescence staining for cartilage-specific proteoglycan (CSPG). After 15 days of culture, CSPG was found in the auricular process but not in the midsection or rostral process. In order to mark the implanted cells and follow their fate, cartilage was labeled in vitro with [3H]thymidine [3H]TdR). Immediately after labeling 20% of the chondrocytes contained [3H]TdR. After culturing for 5 days, 20% of the chondrocytes were still labeled and 10% of the osteogenic cells also contained radioactive label. The labeling index decreased in both cell types with increased duration of culture. Multinucleated clast-type cells did not contain label. Additional cartilages not labeled with [3H]TdR were exposed to between 20000 and 6000 rad of gamma irradiation before ocular implantation. Irradiated cartilage did not hypertrophy or form bone but a fibrous region developed in the midsection. Cells of the host animal were not induced to form bone around the irradiated cartilage. Our studies suggest that fully differentiated chondrocytes of Meckel's cartilage have the capacity to become osteocytes, osteoblasts, and fibroblasts. PMID- 3044882 TI - Diet-induced type II diabetes in C57BL/6J mice. AB - We investigated the effects of diet-induced obesity on glucose metabolism in two strains of mice, C57BL/6J and A/J. Twenty animals from each strain received ad libitum exposure to a high-fat high-simple-carbohydrate diet or standard Purina Rodent Chow for 6 mo. Exposure to the high-fat, high-simple-carbohydrate, low fiber diet produced obesity in both A/J and C57BL/6J mice. Whereas obesity was associated with only moderate glucose intolerance and insulin resistance in A/J mice, obese C57BL/6J mice showed clear-cut diabetes with fasting blood glucose levels of greater than 240 mg/dl and blood insulin levels of greater than 150 microU/ml. C57BL/6J mice showed larger glycemic responses to stress and epinephrine in the lean state than AJ mice, and these responses were exaggerated by obesity. These data suggest that the C57BL/6J mouse carries a genetic predisposition to develop non-insulin-dependent (type II) diabetes. Furthermore, altered glycemic response to adrenergic stimulation may be a biologic marker for this genetic predisposition to develop type II diabetes. PMID- 3044883 TI - Alteration of 1,2-diacylglycerol content in myocardium from diabetic rats. AB - 1,2-Diacylglycerol has been proposed to be a secondary messenger; therefore, in this study we evaluated the amount of 1,2-diacylglycerol in heart tissue from streptozocin-induced diabetic rats and examined the effect of insulin treatment on 1,2-diacylglycerol content. Diabetic rats had lower body and ventricular weights and higher ratios of ventricular to body weight, all of which shifted toward normal values after 4 wk of untreated diabetes followed by 4 wk of insulin treatment. The contents of major phospholipids were significantly depressed in the diabetic rat hearts. In contrast, the triglyceride and cholesterol contents in the myocardium were increased by streptozocin injection and completely normalized by insulin treatment, and glucose levels returned to normal. The 1,2 diacylglycerol content in the myocardium was also significantly elevated in the diabetic rats compared with age-matched controls. Moreover, the 1,2 diacylglycerol content was significantly higher in rats with 4 wk of diabetes than in those with 8 wk of diabetes. Insulin treatment in the diabetic rats, however, did not produce any decrease in 1,2-diacylglycerol content. The results of this study suggest that the development of cardiomyopathy induced by streptozocin injection is associated with a high 1,2-diacylglycerol level, which may result in the activation of protein kinase C. Insulin is one of the agonists that generates 1,2-diacylglycerol in myocytes; however, the relationship between the sustained 1,2-diacylglycerol level and the normalization of diabetes by insulin administration is unclear. PMID- 3044884 TI - Treatment with streptococcal preparation (OK-432) suppresses anti-islet autoimmunity and prevents diabetes in BB rats. AB - We have recently shown that a streptococcal preparation (OK-432) inhibits insulitis and prevents diabetes in nonobese diabetic (NOD) mice, an animal model of insulin-dependent diabetes mellitus (IDDM). We extended this study to another model of IDDM, namely BB rats. Male and female BB rats were injected weekly with 0.2 mg OK-432 i.p. starting from 5 to 6 wk and continuing through 20 or 30 wk of age. The cumulative incidence of IDDM over 20 wk in the OK-432-treated BB rats (4 of 54, 7.4%) was significantly (P less than .01) lower than that found in the nontreated BB rats (13 of 47, 27.7%). We examined some of these rats as follows. All of the OK-432-treated BB rats tested showed normal glucose levels before and after oral glucose administrations, as did the nontreated and nondiabetic BB rats. Histological examination of pancreatic sections revealed that the OK-432 treated rats retained a greater number of intact islets without infiltration of the mononuclear cells than did the nontreated BB rats. A preliminary in vitro study further demonstrated that the cytotoxic activities of spleen cells against a rat insulinoma cell line, RIN, were suppressed in the OK-432-treated rat. However, the treatment of BB rats with OK-432 showed no suppressive effects in the spleen cell number, the responsiveness of spleen cells to concanavalin A, the populations of OX19+, W3/25+, and OX8+ peripheral blood lymphocytes, or in the titers of cell surface antibody against RIN. These results suggest that a nonimmunosuppressive immunomodulator such as OK-432 may be useful as an agent for immunotherapy of IDDM. PMID- 3044885 TI - Reevaluation of urine C-peptide as measure of insulin secretion. AB - Urine C-peptide (UCP) has been proposed as a measure of insulin secretion, because insulin and C-peptide are consecreted in equimolar concentrations by the pancreatic beta-cell. The validity of this approach was tested by comparing insulin secretion rates, calculated by application of a two-compartmental analysis of peripheral C-peptide concentrations, with UCP excretion rates. Insulin secretion and UCP excretion with subjects on a mixed diet were simultaneously measured over a 24-h period in 13 patients with noninsulin dependent diabetes mellitus and in 14 matched nondiabetic control subjects. The fraction of secreted C-peptide that was excreted in the urine (fractional C peptide excretion) showed considerable intersubject variability in the diabetic (11.3 +/- 1.6%, range 3.9-20.8) and control (8.0 +/- 1.7%, range 1.1-27.9, P = .07) subjects (means +/- SE). UCP clearance demonstrated a similar degree of variability and was not significantly different (P = .07) between diabetic (23.8 +/- 3.0 ml/min) and control (16.5 +/- 2.7 ml/min) subjects. In control subjects, the 24-h insulin secretion rate correlated more closely with the fasting insulin secretion rate (r = .97, P = .0001), fasting C-peptide (r = .81, P = .0005), and fasting insulin (r = .80, P = .0005) concentrations than with the 24-h UCP excretion rate (r = .62, P = .02). Similar results were obtained in the diabetic patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3044886 TI - Effect of hyperinsulinemia on urea pool size and substrate oxidation rates. AB - Recently, indirect calorimetry has frequently been used together with hyperinsulinemic clamps. With few exceptions, however, no attention was paid in these studies to the possible effects of hyperinsulinemia on urea nitrogen (N) pool size and the consequences of such changes on the calculated rates of protein, lipid, and carbohydrate (CHO) oxidation. We have determined the effects of euglycemic-hyperinsulinemic clamps on urea N pool size, urinary N excretion, and rates of protein, lipid, and CHO oxidation (measured by indirect calorimetry) in six normal men. Insulin infusion (1 mU.kg-1.min-1) increased peripheral venous insulin concentration from 7 +/- 1.2 (mean +/- SE) to 51 +/- 4 microU/ml. Glucose concentration was clamped at 84 +/- 1.1 mg/dl. Between 0 (preclamp) and 360 min (end of clamp), blood urea N concentration decreased from 17.2 +/- 1.1 to 11 +/- 0.8 mg/dl (P less than .001), and the urea N pool decreased from 604 +/- 41 to 388 +/- 30 mmol (P less than .001). The urea N production rate decreased from 461 +/- 91 (preclamp) to 91 +/- 63 mumol/min during the last 4 h of the clamp (P less than .05). Urinary N excretion remained unchanged (705 +/- 113 vs. 905 +/- 125 mumol/min, NS). Correction of urinary N excretion for insulin-induced reductions in the urea N pool resulted in the following changes in substrate oxidation rates (calculated for the last 4 h of the clamp).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3044887 TI - Inhibition of insulin-stimulated glucose transport by factor extracted from serum of insulin-resistant patient. AB - We report a 31-yr-old nondiabetic male patient with acanthosis nigricans whose hyperinsulinemia and insulin resistance could not be explained by anti-receptor antibodies or by an intrinsic defect of insulin binding to his cells. An acid alcohol extract of the patient's serum contained a factor that inhibited insulin stimulated glucose transport in rat adipocytes. Low levels of the factor could be detected in 9 of 13 unselected patients with non-insulin-dependent diabetes. The factor was heat stable and resistant to treatment with acid, base, and various lytic enzymes. It eluted from a Bio-Gel P-2 column with an apparent molecular weight of 300. The factor also inhibited stimulation of glucose transport in adipocytes by the insulin mimickers hydrogen peroxide and sodium vanadate. In vitro incubation of rat soleus muscles in the presence of the factor resulted in inhibition of insulin-stimulated glucose transport. The factor enhanced 125I labeled insulin binding in both adipocytes and muscle. A preparation of insulin receptors obtained from muscles incubated with serum factor showed increased binding of 125I-insulin to the alpha-subunit of the insulin receptor. Autophosphorylation of the beta-subunit and phosphorylation of exogenous substrate were increased in the receptor preparation obtained from muscles that had been incubated with serum factor. However, the increase in kinase activity was approximately the same as the increase in binding activity. No difference in kinase activity was observed when assayed under conditions in which 125I-insulin binding activity had been equalized.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3044888 TI - Insulin production and glucose metabolism in isolated pancreatic islets of rats with NIDDM. AB - Rats with non-insulin-dependent diabetes mellitus (NIDDM) induced by neonatal injection of streptozocin are known to have a deficient insulin response to glucose. To evaluate to what extent this glucose insensitivity can be attributed to a perturbation of the islet glucose metabolism, we estimated the rates of glucose phosphorylation, glucose utilization, oxygen consumption, and glucose oxidation in islets isolated from normal and NIDDM rats and compared these values with rates of islet insulin biosynthesis and release in vitro. The data confirm that islets from rats with NIDDM display a deficient response to glucose of both insulin biosynthesis and release that is still present after an overnight culture of the islets at 5.5 mM glucose. Furthermore, they show that islets of these rats have 1) normal low- and high-Km glucose-phosphorylating activities and no major alteration of the glucose utilization rate, 2) decreased insulin release in response to glyceraldehyde, 3) decreased rates of basal respiration and glucose oxidation and a markedly reduced stimulation by glucose of both islet oxygen consumption and glucose oxidation, and 4) decreased glucose-stimulated net 45Ca uptake. We conclude that the relative unresponsiveness to glucose of islets from NIDDM rats is associated with, and perhaps due to, a deficient islet glucose metabolism. This defect is not due to gross alterations in the glycolytic pathway but probably reflects alteration in the islet mitochondria function. PMID- 3044889 TI - Insulin-mimetic effects of vanadate in primary cultures of rat hepatocytes. AB - To evaluate possible mechanisms by which insulin inhibits hepatic apolipoprotein B (apoB) secretion, we incubated primary cultures of rat hepatocytes with sodium orthovanadate, a phosphotyrosine phosphatase inhibitor and insulin-mimetic agent. Vanadate (10 microM) and insulin (10 nM) inhibited the medium accumulation of apoB (secretion) by 21 and 37%, respectively, without increasing intracellular apoB. The effects of insulin and vanadate together were not additive. Both insulin and vanadate enhanced intracellular glycogen accumulation by 82 and 37%, respectively. Unlike insulin, vanadate, at a concentration that inhibited apoB secretion (10 microM), had no effect on intracellular lipogenesis, inhibited the secretion of newly synthesized hepatic proteins, and had a delayed onset and termination of action on inhibition of apoB secretion. At higher concentrations (40 and 80 microM), vanadate stimulated intracellular lipogenesis. In conclusion, our data indicate that vanadate mimics insulin action in hepatocytes with regard to the inhibition of medium accumulation of apoB. These data are consistent with the hypothesis that inhibition of apoB secretion may be secondary to an increase in phosphotyrosine content at its site of synthesis. The kinases responsible for this effect have not been identified. Several effects of vanadate, however, are different from those of insulin, suggesting a differential sensitivity to vanadate, a divergence of the signal transfer by insulin and vanadate at the insulin-receptor or postreceptor level, or both. PMID- 3044890 TI - Long-term follow-up of polyneuropathy in diabetic kidney transplant recipients. AB - Nerve conduction and electromyography (EMG) of insulin-dependent (type 1) diabetic patients with end-stage nephropathy was studied before and up to 10 yr after kidney transplantation (KTx). A series of nondiabetic KTx patients served as a comparison group. Motor nerve conduction velocity (NCV) was measured in the ulnar, median, peroneal, and tibial nerves; sensory NCV was measured in the median nerve. EMG was performed in the first dorsal interosseus, flexor carpi radialis, anterior tibialis, and gastrocnemius muscles. In 68 pre-KTx diabetic patients, the mean NCV was below normal in all nerves, and the mean amplitudes of the evoked muscle action potential (MAP) were low normal in the upper extremity and below normal in the lower extremity. The values of the comparison group were within the normal range. At 1 (n = 57), 5 (n = 23), and 10 (n = 10) yr after KTx, the mean NCV of the diabetic patients remained essentially unchanged, but MAP amplitudes of all muscles had declined. EMG revealed progression of the denervation process, especially in muscles of the lower extremities. We conclude that diabetic neuropathy continues to progress by a progressive axonal loss after correction of uremia by KTx. PMID- 3044891 TI - Effect of vitamin E supplementation on platelet thromboxane A2 production in type I diabetic patients. Double-blind crossover trial. AB - Vitamin E deficiency is associated with increased platelet aggregation, which can be normalized through vitamin E supplementation. In diabetes, increased platelet thromboxane A2 (TXA2) production is correlated with decreased platelet vitamin E content. We therefore investigated the effect of 400 mg DL-alpha-tocopherol acetate daily for 4 wk on ADP- and collagen-induced platelet aggregation and platelet TXA2 production in 22 type I (insulin-dependent) diabetic patients without macroangiopathy and with no or only minimal microangiopathy by a double blind placebo-controlled crossover study. Platelet aggregation was induced in platelet-rich plasma by two or three different concentrations of ADP and collagen. TXA2 was measured by the stable spontaneous breakdown product thromboxane B2 by a specific radioimmunoassay. Whereas metabolic control remained unchanged during the study period, platelet TXA2 production was significantly (P less than .05 and P less than .01) reduced at each ADP concentration and at two of three collagen concentrations. Because increased TXA2 production of diabetic platelets is thought to play an important pathogenetic role in diabetic angiopathy, we conclude that vitamin E treatment could be beneficial with respect to platelet-vessel-wall interaction and thus might be promising for the prevention of diabetic angiopathy. PMID- 3044892 TI - Interrelationships among insulin's antilipolytic and glucoregulatory effects and plasma triglycerides in nondiabetic and diabetic patients with endogenous hypertriglyceridemia. AB - We tested the hypothesis that the previously observed association among hypertriglyceridemia, hyperinsulinemia, and insulin resistance could be explained by a defect in insulin's antilipolytic effect. Insulin action was measured in 10 nondiabetic and 8 diabetic patients with hypertriglyceridemia (fasting plasma triglyceride 800 +/- 154 and 1105 +/- 445 mg/dl, respectively, P NS; fasting plasma glucose 99 +/- 3 and 161 +/- 12 mg/dl, respectively, P less than .001) and in 8 weight-matched normolipemic nondiabetic individuals (fasting plasma triglyceride and glucose 110 +/- 21 and 91 +/- 3 mg/dl). The slope of the decay in plasma free fatty acid (FFA) during insulin infusion was used as an index of insulin's antilipolytic effect. Insulin stimulation of glucose uptake in vivo during intravenous hyperinsulinemic clamp and in vitro in adipocytes were measures of insulin's glucoregulatory action. Both glucoregulatory and antilipolytic effects were similarly reduced in both hypertriglyceridemic groups compared with normal subjects. The plasma triglyceride concentration correlated positively with the slope of FFA suppression by insulin (r = .81, P less than .0001) and the fasting FFA concentration (r = .65, P less than .0001). In multiple linear regression analysis, insulin's antilipolytic effect and the fasting FFA concentration explained 83% of the variation in the plasma triglyceride concentration. These associations were independent of insulin's glucoregulatory effect and the fasting plasma insulin concentration. The data indicate that patients with endogenous hypertriglyceridemia are resistant to both the antilipolytic and glucoregulatory actions of insulin and that increased flux of FFA as a result of the latter, rather than hyperinsulinemia, is responsible for elevation of very-low-density lipoprotein production.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3044894 TI - Distinct macrophage subpopulations in pancreas of prediabetic BB/E rats. Possible role for macrophages in pathogenesis of IDDM. AB - Use of monoclonal antibodies directed against rat macrophages and serial pancreatic biopsy in the prediabetic period have enabled us to document the involvement of macrophages in the pancreatic events leading to onset of diabetes in the spontaneously diabetic BB/E rat. A few weeks before onset of disease, there is marked recruitment and accumulation of ED1+ macrophages at periductal and perivascular locations adjacent to noninfiltrated islets. These recruited cells, distinct from the resident ED2+ tissue macrophages, then infiltrate the islets. Infiltration of the pancreas by ED1+ macrophages is therefore a very early event in the prediabetic period and suggests a possible role for macrophages in the pathogenesis of insulin-dependent diabetes mellitus (IDDM) in this animal model. PMID- 3044893 TI - Genetic control of organ-reactive autoantibody production in mice by obesity (ob) diabetes (db) genes. AB - Inbred strains of mice exhibited genetic and sex-dependent differences in spontaneous production of organ-reactive autoantibodies detected by indirect immunofluorescence. Antitestis autoreactivity was found primarily in sera from C57BL/6J (B6) mice, whereas antigastric autoreactivity was common to both CBA/J and 129/J strains. Autoantibodies against islet cell cytoplasmic antigens (ICAs) were uniquely expressed by C57BL/KsJ (BKs) males. Introduction of the diabetes (db) mutation into these various inbred-strain backgrounds induced expression of ICA, with stronger induction observed in males. The stress imposed by the db or obesity (ob) mutation induced ICA in BKs mice at a higher frequency than in B6 mice; this differential sensitivity was somehow related to a gene linked to the H 2 complex because BKs.B6 H-2b congenic mice resembled B6 mice. The db3J mutation increased the expression of these autoantibodies in 129/J mice, which, like B6, were H-2b and therefore presumably possessed the same H-2-linked inducibility allele as BKs. Cytotoxic autoantibodies against islet cell surface antigens were only observed in C3HeB/FeJ db/db males, and their presence was correlated with beta-cell necrosis. It is concluded that db and/or ob genes appear to play an important role in the production of autoantibodies to islet cells, and sex-linked factor(s) may modify the phenotypic expression of the autoantibodies. PMID- 3044895 TI - Adoptive immunotherapy of diabetes in autologous nonobese diabetic mice with lymphoid cells ex vivo exposed to cyclosporin plus interleukin 2. AB - Between 12 and 26 wk of age, approximately 80% of female nonobese diabetic (NOD) mice from the inbred Sansum colony develop lymphocytic insulitis and become overtly diabetic. The disease in this animal model is similar to human insulin dependent diabetes mellitus (IDDM) in both genetics and autoimmune pathogenesis. Cyclosporin A (CsA) has been used for immunosuppression in IDDM of recent onset in humans but has several limiting side effects. Therefore, a different regimen for CsA immunosuppression was investigated. Autologous splenic lymphoid cells from 12-wk-old not-yet-diabetic female NOD mice were cultured for 72 h with CsA plus interleukin 2 (IL-2) before reinfusion into the animal from which they were isolated. After this treatment, only 2 (18%) of 11 mice became overtly diabetic during an observation period of 19 wk, while 18 (86%) of 21 age-matched control mice developed diabetes during the same observation period (P less than .001). These data suggest an ex vivo preferential IL-2 activation of specific suppressor cells for the autoimmune process with CsA blockade of cytolytic/helper activities. Because the in vivo concentrations of CsA with this procedure would be negligible, these findings have implications for the potential nontoxic use of CsA in human protocols as well. PMID- 3044896 TI - The achlorhydria-carcinoid sequence: role of gastrin. PMID- 3044897 TI - Digital filtering of auditory evoked potentials. AB - Digital filters have been proposed by many authors for analysis and enhancement of the auditory brain stem response and related potentials. Because these techniques are unfamiliar to many, and require attention to underlying assumptions, an overview of the field is presented, with emphasis on limitations and risks of each method. Topics include Fourier transforms, finite impulse response (convolutional) filters, and infinite impulse response (recursive) filters. PMID- 3044898 TI - Hypo- and hyperthermia in clinical auditory brain stem response measurement: two case reports. AB - Two case reports are presented to highlight the important effects of body temperature in clinical auditory brain stem response (ABR) measurement. Case 1 is an 11 year old boy in coma secondary to severe head injury. High dose barbiturate therapy suppressed brain stem neurologic signs and the ABR was relied on as a monitor of CNS status. Hypothermia during this period of intensive care was a crucial factor for meaningful interpretation of ABR findings. The second case was a 26 year old male undergoing hyperthermic therapy for advanced cancer. As body temperature increased from 38 to 42 degrees Centigrade (107.6 degrees Fahrenheit), there was a systematic decrease in latency for waves III and V. An overall hyperthermia-related decrease in the wave I-V latency interval of 0.5 to 0.6 milliseconds was observed on two test dates. ABR results for these two cases are discussed in the context of basic knowledge on body temperature and auditory electrophysiology. PMID- 3044899 TI - [Problems in the physiology of the human brain (its status and outlook)]. PMID- 3044900 TI - [The problem of the chemical asymmetry of the brain]. PMID- 3044901 TI - [Current concepts of the psychophysiological significance of P300]. PMID- 3044902 TI - Luis Federico Leloir. September 6, 1906-December 2, 1987. PMID- 3044903 TI - Disruption of hepatic heme biosynthesis after interaction of xenobiotics with cytochrome P-450. AB - Heme biosynthesis in hepatocytes is controlled by a free heme pool, which regulates delta-aminolevulinic acid synthase. Porphyrinogenic chemicals deplete the regulatory free heme pool by interacting with cytochrome P-450 thereby inhibiting heme biosynthesis and/or causing heme breakdown. Recent developments allow us to predict which groups of chemicals are likely to be porphyrinogenic. One group is exemplified by 3,5-diethoxycarbonyl-1,4-dihydro-2,4,6 trimethylpyridine. Heterocyclic compounds of this type cause mechanism-based inactivation of cytochrome P-450, leading to the formation of N-alkylporphyrins, with ferrochelatase-inhibitory activity resulting in lowering the free heme pool. Allylisopropylacetamide exemplifies a second group. Such compounds containing a terminal olefinic or acetylenic group, cause mechanism-based inactivation of cytochrome P-450. In the process, the heme moiety of cytochrome P-450 is destroyed and the free heme pool is lowered. A third group is exemplified by planar polyhalogenated or polycyclic aromatic hydrocarbons. These compounds induce specific cytochrome P-450 isozymes but are poor substrates. Active oxygen is formed, which interacts with a hepatic substrate to form a uroporphyrinogen decarboxylase inhibitor. Inhibition of this enzyme leads to depletion of the free heme pool. PMID- 3044904 TI - Resistance of multidrug-resistant lines to natural killer-like cell-mediated cytotoxicity. AB - Multidrug resistance (MDR) refers to a complex phenotype that describes a number of features characterized primarily by resistance to a wide range of structurally unrelated drugs. In this paper we investigated the relationship between drug resistance and resistance to NK-mediated cytotoxicity. Studies with two independently selected multidrug-resistant cell lines indicated that increased drug resistance was associated with both an increased resistance to NK-mediated cytotoxicity and increased levels of membrane P-glycoprotein expression. This resistance to cytotoxicity appears to result partly from an alteration in the membrane structure of the target cells inasmuch as there was a reduction in effector:target cell recognition. Resistance to NK-mediated cytotoxicity should be included with the numerous pleiotropic changes associated with the multidrug resistance phenotype. PMID- 3044905 TI - Aging and glucose homeostasis in C57BL/6J male mice. AB - Age-dependent changes in glucose homeostasis were assessed in specific pathogen free C57BL/6J male mice. Increased islet size and pancreatic insulin content in old (21-25-month-old) mice were associated with lower nonfasting plasma glucose levels and improved clearance of either an oral or an i.p. administered glucose load in comparison with young, mature (4-5-month-old) males. The almost twofold increase in islet size correlated with a twofold increase of glucose-stimulated insulin secretion from perifused islets from 25-month-old males compared with 5 month-old males. These aging male mice did not become obese, and there were no fibrotic changes associated with the hyperplastic islets observed in the old males. Thus, the findings that glucose tolerance did not deteriorate with age, coupled with the lack of evidence for impaired beta cell responsiveness to glucose in old males, suggest that deterioration in glucose homeostasis is not an inevitable consequence of aging in the mouse. PMID- 3044907 TI - [Left ventricular thrombosis after myocardial infarct]. PMID- 3044906 TI - Production of immunoreactive growth hormone by mononuclear leukocytes. AB - In the present study, we evaluated whether mononuclear leukocytes could synthesize and secrete growth hormone (GH) in vitro. By using RNA slot blot analysis, we detected maximum spontaneous levels of specific GH mRNA in the cytoplasm of rat leukocytes after a 4-h incubation. Northern gel analysis demonstrated that the specific leukocyte GH RNA was polyadenylated and had a molecular mass of 1.0 kb. Further studies using immunofluorescence, antibody affinity chromatography, and Sephacryl gel filtration indicate that leukocytes secrete a high molecular weight (greater than 300,000) and a low molecular weight (approximately 22,000) immunoreactive GH (irGH). A substantial amount of the high molecular weight irGH can be converted to the lower molecular weight form after reduction with mercaptoethanol. The irGH appeared to be de novo synthesized because it could be radiolabeled with tritiated amino acids and its production could be blocked by previous incubation of leukocytes with cycloheximide. The replication of Nb2 rat node lymphoma cells was stimulated by affinity-purified human lymphocyte-derived irGH. The growth stimulation was blocked by specific antibodies to hGH. We conclude that lymphocytes produce an irGH that is similar to if not identical to pituitary GH in terms of bioactivity, antigenicity, and molecular weight. The findings demonstrate a potential regulatory loop between the immune and neuroendocrine tissues. PMID- 3044908 TI - [Diltiazem in the treatment of angina pectoris]. PMID- 3044909 TI - Do skin tags constitute a marker for colonic polyps? A prospective study of 100 asymptomatic patients and metaanalysis of the literature. AB - Several reports have suggested that skin tags may be a marker for the presence of colonic polyps in symptomatic patients referred for colonoscopy. In a prospective study of 100 asymptomatic patients, we found no association between skin tags and colonic polyps. A review of the literature and results of a metaanalysis show a significant association between skin tags and colonic polyps in 777 symptomatic patients, but no association in 268 asymptomatic patients. To explain this discrepancy, several possible biases are analyzed. As skin tags constitute a marker for colonic polyps only in symptomatic patients for whom a colonoscopy is already indicated, their detection is of no diagnostic value in asymptomatic patients. PMID- 3044910 TI - Physiologic significance and regulation of hepatocellular heterogeneity. PMID- 3044911 TI - Sludge, stones, and pregnancy. PMID- 3044912 TI - Healing and relapse of severe peptic esophagitis after treatment with omeprazole. AB - We have studied the response of erosive or ulcerative esophagitis to treatment with omeprazole and its subsequent relapse on cessation of therapy in 196 patients. In the first phase of the study omeprazole (20 or 40 mg daily) was compared with placebo in 64 patients. After 4 wk there was endoscopic healing in 81% (25 of 31) of omeprazole-treated patients and in only 6% (2 of 32) of placebo treated patients. Endoscopic healing of esophagitis was accompanied by symptom relief and histologic healing of ulceration. In the second (dose finding) phase a further 132 patients were randomized to omeprazole (20 or 40 mg daily) and endoscopic healing was assessed. In patients with the mildest grade of ulcerative esophagitis (grade 2), healing occurred at 4 wk in 87% receiving 20 mg and in 97% receiving 40 mg. In patients with grade 3 esophagitis, 67% (20 mg) and 88% (40 mg) were healed. Less than half the patients with grade 4 esophagitis (Barrett's ulcers or confluent ulceration) healed with either 20 mg (48%) or 40 mg (44%). Regression analysis in the 164 omeprazole-treated patients showed no evidence that healing was influenced by factors other than severity of esophagitis at entry and omeprazole dose. In phase 3 of the study the rate of endoscopic relapse was determined in 107 endoscopically healed patients after stopping omeprazole. Erosive or ulcerative esophagitis recurred in 88 of 107 (82%) by 6 mo. Neither initial dose, grade of esophagitis, nor smoking was shown to influence relapse rate. Omeprazole is a highly effective treatment for peptic esophagitis. The 40 mg/day dosage produces endoscopic healing slightly more quickly than the 20 mg/day dosage, and the initial endoscopic gradings are of prognostic value. Relapse occurs rapidly when treatment is stopped. PMID- 3044915 TI - Prophylactic injection sclerotherapy for esophageal varices: a critical appraisal. PMID- 3044913 TI - Endoscopic sclerotherapy versus medical treatment for bleeding esophageal varices in patients with schistosomal liver disease. AB - A prospective controlled trial was conducted at Ain-Shams and Benha University Hospitals. One-hundred and eighteen chronic liver disease patients, mostly schistosomal in origin and presenting with recent proven variceal hemorrhage, were randomly allocated to injection sclerotherapy or medical therapy. The follow up period extended to 21 months. Sixty-three patients received injection sclerotherapy using ethanolamine oleate (5% wt/vol) paravariceally while 55 received medical treatment in the form of general resuscitative measures, blood transfusion, vasopressin intravenous drip, and insertion of a Sengstaken Blakemore tube if bleeding continued. The first 30-day mortality was 7 (11%) in the injection sclerotherapy group compared with 11 (20%) in the medical treatment group. This difference was not statistically significant. During the entire observation period 9 (14.3%) died in the sclerosed group and 16 (29%) died in the medically treated group, and this difference was statistically significant at the 5% level. Comparison of recurrent bleeding among both groups revealed that the difference was statistically not significant. It was concluded that injection sclerotherapy was no better than medical treatment in the control of acute variceal bleeding, but injection sclerotherapy did increase significantly long term survival of sclerosed patients. PMID- 3044914 TI - Consecutive Serratia marcescens infections following endoscopic retrograde cholangiopancreatography. PMID- 3044916 TI - [The solid-phase immunoenzyme method of determining the ferritin concentration of human blood serum]. PMID- 3044917 TI - Actions of avermectins on cultured neurons. PMID- 3044918 TI - Domperidone. PMID- 3044919 TI - Induction of ovulation and oviposition in female quail with luteinizing hormone, luteinizing hormone releasing hormone, or progesterone. AB - Regularly laying female Japanese quail were injected 12 or 18 hr before the next expected oviposition with 25 micrograms oLH, 25 micrograms luteinizing hormone releasing hormone (LHRH), or 0.05 mg progesterone, and the subsequent oviposition was recorded and ovulation determined by autopsy 9 hr after the injection. Plasma progesterone levels were measured in blood collected from a wing vein during the postinjection interval. Vehicle-injected birds served as control. All treatments resulted in premature oviposition and ovulations in 50-100% of birds when injected 12 hr before the next expected oviposition. None of the vehicle-injected birds showed any advancement in either oviposition or ovulation. Premature oviposition was generally followed by premature ovulation within 1 hr, when birds were treated 12 hr before the next expected oviposition and was preceded by a rise in plasma progesterone levels which reached values similar to those occurring during the normal preovulatory period. When birds were injected 18 hr before the next expected oviposition, the incidence of premature oviposition was very low, premature ovulation was absent, and the rise in plasma progesterone levels following treatment was substantially less than in the former group. The results suggest that oviposition and ovulation in quail may be initiated by LHRH induced LH release from the pituitary gland and that progesterone may stimulate LHRH and LH release. The timing of the ovulatory cycle may depend upon the phase of follicular maturation. PMID- 3044921 TI - A diamond in a desert. PMID- 3044920 TI - Photoperiodic control of puberty in the red-legged partridge (Alectoris graeca chukar). AB - The photoperiodic control of puberty in birds was studied using hybrid red-legged partridges (Alectoris graeca chukar). In the wild partridges first breed at about 10 months of age. Males and females were held from hatch on short (8L:16D) daylengths and then at 3, 4 1/2, 6, 7 1/2, 9, and 12 weeks of age groups were transferred to long daylengths (20L:4D) for 3 weeks to test for the photoperiodic induction of gonadal growth. Control groups were maintained on either the short or long daylengths and killed at 3, 9, and 12 weeks of age. Gonadal growth did not occur in either of the control groups, nor in any of the birds photostimulated prior to 6 weeks of age. However, in birds photostimulated after this point gonadal size was increased in a graded manner, this growth being reflected in the plasma and pituitary levels of luteinizing hormone. Similar long day responses which increased with age were observed in plasma and pituitary levels of prolactin (PRL). In the control birds there was a steady decline with time in plasma PRL, with no change being seen in pituitary PRL. Hypothalamic gonadotrophin-releasing hormone content did not change in birds maintained on 8L:16D or 20L:4D and was only elevated by photostimulation in the females. Plasma testosterone remained low in all the males except for those transferred to 20L:4D at 12 weeks. Testicular testosterone content remained low in control birds but was elevated by photostimulation at 6 and 12 weeks of age. Plasma progesterone levels increased with age in control birds and were only elevated by photostimulation at 3 and 4 1/2 weeks of age. These data show juvenile partridges to be completely unresponsive to photoperiodic stimulation and in a condition very similar if not identical to that seen in adult refreactoriness. As in the adults, this refractoriness is progressively dissipated by exposure to short days so that by 9-12 weeks of age the partridges are able to respond maximally to long day stimulation. PMID- 3044922 TI - Double-stranded gap repair of DNA by gene conversion in Escherichia coli. AB - We demonstrated repair of a double-stranded DNA gap through gene conversion by a homologous DNA sequence in Escherichia coli. We made a double-stranded gap in one of the two regions of homology in an inverted orientation on a plasmid DNA molecule and introduced it into an E. coli strain which has the RecE system of recombination (genotype; sbcA23 recB21 recC22). We detected repair products by genetic selection. The repair products were those expected by the double-strand gap repair model. Gene conversion was frequently accompanied by crossing over of the flanking sequences as in eukaryotes. This double-strand gap repair mechanism can explain plasmid recombination in the absence of an artificial double-stranded break reported in a companion study by Yamamoto et al. PMID- 3044923 TI - Genetic control of intrachromosomal recombination in Saccharomyces cerevisiae. I. Isolation and genetic characterization of hyper-recombination mutations. AB - Eight complementation groups have been defined for recessive mutations conferring an increased mitotic intrachromosomal recombination phenotype (hpr genes) in Saccharomyces cerevisiae. Some of the mutations preferentially increase intrachromosomal gene conversion (hpr4, hpr5 and hpr8) between repeated sequences, some increase loss of a marker between duplicated genes (hpr1 and hpr6), and some increase both types of events (hpr2, hpr3 and hpr7). New alleles of the CDC2 and CDC17 genes were recovered among these mutants. The mutants were also characterized for sensitivity to DNA damaging agents and for mutator activity. Among the more interesting mutants are hpr5, which shows a biased gene conversion in a leu2-112::URA3::leu2-k duplication; and hpr1, which has a much weaker effect on interchromosomal mitotic recombination than on intrachromosomal mitotic recombination. These analyses suggest that gene conversion and reciprocal exchange can be separated mutationally. Further studies are required to show whether different recombination pathways or different outcomes of the same recombination pathway are controlled by the genes identified in this study. PMID- 3044924 TI - An efficient selection producing structural gene mutants of yeast alcohol dehydrogenase resistant to pyrazole. AB - Selection for resistance to allyl alcohol in respiration-incompetent Saccharomyces cerevisiae produces a high proportion of mutants that can be localized within the ADH2 structural gene and that still, because of the type of selection employed, retain enzyme activity. We show here that a similar type of selection produces a similarly high proportion of mutants resistant to the competitive inhibitor pyrazole. The first four mutants examined, picked at random from a collection of spontaneous pyrazole-resistant mutants, show altered- usually increased--KM values for ethanol and NAD+, and markedly increased K1 values for pyrazole, compared with the wild type. When these kinetic measures and their electrophoretic mobilities were compared, all the mutants could be clearly distinguished from each other as well as from wild type. Genetic analysis shows these mutants to be close to and probably resident in the structural gene. For a variety of reasons, these mutants are even more favorable subjects for population genetic analysis and the dissection of molecular microevolution than are allyl alcohol-resistant mutants. PMID- 3044925 TI - Acquisition of new metabolic capabilities: multicopy suppression by cloned transaminase genes in Escherichia coli K-12. AB - The four general transaminases of Escherichia coli K-12 have overlapping, but discrete, substrate specificities and participate in the final step in the synthesis of at least seven different amino acids. Through the use of strains that have mutations in one or more transaminase genes and carry a different wild type (wt) gene on a multicopy plasmid, it was possible to detect instances in which an amplified wt gene suppressed nonallelic mutations. In these cases, overproduction of the enzyme permitted a broader range of substrates to be used at physiologically significant levels, either because a low catalytic efficiency (in the case analyzed here) or a low affinity of the enzyme towards the substrate prevented its effective utilization under normal conditions. Consequently, by compensating for a low catalytic reaction rate, enzyme overproduction circumvents the original lesion and restores biosynthetic activity to the mutant strain. The suppression of a mutation in one gene by amplified copies of a different wt gene is termed 'multicopy suppression'. This phenomenon is useful for detecting poorly expressed genes, for detecting duplicate genes, for identifying secondary functions of the products of known genes, and for elucidating the metabolic role of the product of the suppressed gene. PMID- 3044926 TI - Synthesis of functional mouse cytochromes P-450 P1 and chimeric P-450 P3-1 in the yeast Saccharomyces cerevisiae. AB - Mouse liver cytochrome P-450 P1 was produced in the yeast Saccharomyces cerevisiae transformed by various expression vectors. The relative efficiency of the phosphoglycerate kinase and GAL10-CYC1 promoters to direct the P-450 P1 mRNA synthesis was determined. The level of protein synthesis was found to be dependent on the amount of the 5'-noncoding sequence of the original cDNA removed during the construction. Yeast-synthesised P-450 P1 was found to be integrated into the microsomal membrane in a fully functional form, as judged by Western blotting, optical spectra and enzymatic activities. The amount of P-450 reached up to 0.6% of the microsomal protein level. A nucleotide sequence coding for a chimeric enzyme in which 40 N-terminal codons of P-450 P1 were replaced by 36 N terminal codons of P-450 P3 was constructed and expressed in yeast. The resulting protein retained full P-450 P1 activity and was produced with a similar efficiency suggesting that the P-450 N-terminal sequence is not involved in structures critical for the substrate specificities of the P1 isoenzyme. PMID- 3044929 TI - Influenza vaccination in the elderly: can efficacy be enhanced? AB - Influenza remains a major cause of illness and death in geriatric populations, especially in the frail elderly and those who live in nursing homes. Influenza vaccine has been useful in reducing morbidity and mortality; however, the vaccine is not uniformly successful in preventing infection, particularly in those people who are immunocompromised. In this article, the rationale and expectations for the current vaccine are reviewed and strategies to enhance vaccine efficacy are presented. PMID- 3044928 TI - Cancer in the older woman: diagnosis and prevention. AB - Progestogens should be added to estrogen replacement therapy, not only to prevent endometrial cancer in women with a uterus, but also to reduce the risk of breast cancer in some women. Smoking should be discouraged to reduce the risk for both lung cancer and heart disease. Recommendations should be made to increase fiber intake to lessen the risk for carcinoma of the colon. Reducing fat intake also decreases risk for colon cancer, as well as carcinoma of the breast. Postmenopausal bleeding must be investigated for early diagnosis of endometrial cancer and, when endometrial hyperplasia is the finding, it should be treated with progestogens to prevent adenocarcinoma. The progestogen challenge test is recommended for all women with a uterus, and if bleeding occurs, the progestogen should be continued for 13 days each month. Use of mammograms and other diagnostic modalities should be increased to make the earliest possible diagnosis of breast cancer. PMID- 3044927 TI - Nucleotide sequence and expression in Escherichia coli of cDNA of swine pepsinogen: involvement of the amino-terminal portion of the activation peptide segment in restoration of the functional protein. AB - A clone, pSPcA2, which carries the full-length swine pepsinogen cDNA was isolated. The coding sequence comprised the signal peptide [15 amino acids (aa)], the activation peptide segment (44 aa) and mature pepsin (327 aa). The deduced amino acid sequence agrees with the published sequence with two exceptions. Asparagine instead of aspartate is present at aa positions 19 and 308. Two types of plasmids, pAS and pUCtacSPc series, were constructed for expressing swine pepsinogen cDNA. These plasmids directed the synthesis of polypeptides which were detected by employing an antibody to swine pepsinogen. However, all the polypeptides formed aggregates and showed no acid protease activity. Only the protein directed by pAS5 regained the acid protease activity after renaturation procedures. The activity was completely inhibited by pepstatin. Furthermore, the renatured pAS5 protein was spontaneously converted to pepsin under acidic conditions. The presence of Arg-8 in the activation peptide segment appears important for the stabilization of the pepsinogen molecule. PMID- 3044930 TI - Mycobacteria and Crohn's disease. PMID- 3044931 TI - Regulation of glucose homeostasis in rat jejunum by despentapeptide-insulin in vitro. AB - The regulation of the absorption and metabolism of glucose in rat small intestine by insulin was studied by the perfusion of isolated loops of proximal jejunum in vitro. The addition of an active, monomeric form of insulin, despentapeptide insulin, to the serosal side of the intestine from normal rats inhibited luminal glucose absorption (421 (11) to 285 (11) mumol/h/g dry wt, p less than 0.001) and lactate production (340 (28) to 192 (26) mumol/h/g dry wt, p less than 0.001), but had no effect on glucose utilisation (231 (11) and 210 (16) mumol/h/g dry wt). The production of acute insulin deficiency by the injection of anti-insulin serum in vivo caused a marked inhibition of luminal glucose absorption (421 (11) to 240 (13) mumol/h/g dry wt, p less than 0.001), glucose utilisation (231 (11) to 48 (2) mumol/h/g dry wt, p less than 0.001) and lactate production (340 (28) to 94 (2) mumol/h/g dry wt, p less than 0.001) in vitro. The effects of insulin deficiency were reversed by despentapeptide-insulin in vitro, so that the rates of absorption and metabolism for intestine from insulin deficient and normal rats were similar in the presence of the modified insulin. All the effects caused by insulin deficiency and despentapeptide-insulin were apparent within minutes and could not be attributed to hyperglycaemia. It is concluded that rat small intestine is subject to rapid and direct regulation by insulin. PMID- 3044934 TI - Crohn's or poisoning? PMID- 3044932 TI - Pharmacological constriction of the lower oesophageal sphincter: a simple method of arresting variceal haemorrhage. AB - The effect of pharmacological constriction of the lower oesophageal sphincter (LOS) on oesophageal varices was investigated in an experimental study followed by a controlled clinical trial. In the experimental study intravariceal pressure was measured just above the LOS in 11 patients before and after constricting the LOS by intravenous pentagastrin. Intravariceal pressure fell from a mean of 23 (range 12-36) mmHg to 4 (range 0-7) mmHg (p less than 0.001). This marked pressure drop indicated the considerable compression of varices that occurred within the LOS. A prospective controlled clinical trial examined whether LOS constriction (effected by the longer acting metoclopramide) would compress varices sufficiently to arrest active variceal bleeding originating from the lowest 2 cm oesophagus--the area encircled by the LOS. Of 11 patients who received metoclopramide, 10 stopped bleeding compared with four of the 11 who received placebo (p less than 0.01). Pharmacological constriction of the LOS appears to offer a new and effective approach for arresting active bleeding from oesophageal varices. PMID- 3044935 TI - Breast cancer detection: age-related significance of findings on physical exam and mammography. AB - Data of 909 consecutive patients who underwent physical exam (PE), mammography (MG), and breast biopsy were analyzed in a retrospective study. Preoperative findings of PE and MG were classified as (1) suspicious for malignancy, (2) probably benign lesion, (3) normal and correlated with histology of the biopsy and the patient's age. Sensitivity of PE remained on a 0.9 level approximately up to age 50 and dropped to 0.78 and 0.85 respectively thereafter. Sensitivity of MG was slightly lower and was 0 in the age group 30 years and younger. Specificity of PE rose from 0.52 to 0.84 and dropped to 0.55 in women above 70 years. Specificity of MG decreased from 0.9 to 0.4 over the age groups. The relatively low positive predictive value of both PE and MG--particularly in the lower age groups--leads to an excessive rate of benign biopsies up to the age of 60. Due to suboptimal sensitivity PE and MG have to be used as a combined diagnostic modality if a reduction of breast cancer mortality is to be accomplished. Screening programs are of particular efficacy in women over 45 years of age due to age-dependent incidence figures and diagnostic sensitivity of PE and MG in this age group. PMID- 3044933 TI - Lymphoepithelial interactions in the mucosal immune system. PMID- 3044936 TI - Urinary gonadotropin fragments (UGF) in cancers of the female reproductive system. I. Sensitivity and specificity, comparison with other markers. AB - UGF is a mixture of human chorionic gonadotropin free beta-subunit and its fragments, and is detected in pregnancy and trophoblast disease urines. An examination of 67 nonpregnant cancer-free women showed average urine levels of 0.13 ng/ml. Six of the 67 (8.9%) had levels exceeding a selected cutoff value, 0.2 ng/ml. Of 112 woman with active gynecologic cancer, 72 (64%) had urine UGF levels exceeding this cutoff value. When urines were limited to those with creatinine greater than 0.5 mg/ml, or to first morning samples (mean creatinine 1.0 ng/ml), the sensitivity of UGF for all gynecologic cancers was raised to 76%. The sensitivity of UGF for cervical (73%), for endometrial (65%), and for ovarian (83%) cancers exceeded that of plasma CA 125 and lipid-associated sialic acid in plasma (LASA) in the same population. UGF is an exciting new tumor marker which warrants further evaluation. PMID- 3044937 TI - Long-term cryopreservation of autologous bone marrow: analysis of granulocyte macrophage progenitor (CFU-GM) viability in 31 samples stored more than 48 months. PMID- 3044938 TI - [Ambulatory oocyte retrieval for in vitro fertilization by transvaginal sonography]. PMID- 3044939 TI - [The role of calcium in brain trauma and ischemia]. PMID- 3044940 TI - [Immunotherapy for septic shock]. PMID- 3044941 TI - [Radiation enteritis]. PMID- 3044942 TI - [Autoantibodies in malignant disorders]. PMID- 3044943 TI - [Popliteal cyst and pseudothrombophlebitis]. PMID- 3044945 TI - [The artificial heart: aim or transition to heart transplantation]. PMID- 3044944 TI - [Heart transplantation for hypoplastic left heart syndrome]. PMID- 3044946 TI - Isolation of alkaloids and glycosides from tissue following enzymic digestion. AB - Four enzymic digestion methods have been evaluated for their efficiency in releasing certain alkaloids and glycosides from spiked liver tissue. Enzymic digestion gives better recoveries of all the plant poisons studied than those obtained by conventional methods. A flow diagram for the enzymic methods of drug isolation and quantitation by HPLC is presented. Enzymic digestion for the release of glycosides is reported for the first time. It is concluded that papain digestion is the most suitable method for the analysis of broad spectrum of compounds of forensic and clinical importance. PMID- 3044948 TI - [Sonographic diagnosis of malignant melanoma and its regional lymph drainage in the area of the head and neck]. PMID- 3044947 TI - [So-called juvenile nasopharyngeal fibroma]. PMID- 3044949 TI - [Prognostically relevant criteria and stage-related therapy of malignant melanoma]. PMID- 3044950 TI - [Covering the defect following melanoma surgery of the face]. PMID- 3044951 TI - [Clinical aspects and immunohistochemistry of soft tissue sarcomas of the maxillofacial area]. PMID- 3044952 TI - [Spontaneous remission of malignant melanomas]. PMID- 3044953 TI - [Kaposi sarcoma of the gingiva in an HIV-negative transplantation patient]. PMID- 3044954 TI - [Angiomyoma in the area of the mouth, jaw and face]. PMID- 3044955 TI - [Adult rhabdomyoma]. PMID- 3044956 TI - [Neurofibrohemangiomatous soft tissue changes with pathognomonic mandibular deformity]. PMID- 3044957 TI - The in vivo role of experimentally produced anti-islet-cell-surface-antibodies in the development of diabetes mellitus. PMID- 3044959 TI - RVS will bring changes in hospital utilization. PMID- 3044958 TI - Pharmacological perturbation of cholinergic systems: applications to the study of pathophysiological mechanisms in affective disorders. AB - Clinical, behavioral, physiological, biochemical and receptor binding parameters are useful as dependent measures in studies into the pathophysiology of affective disorders. Pharmacological perturbation of cholinergic mechanisms and modification of cholinergic-monoaminergic interaction mimics aspects of the neurobiology of affective disorders. The effects of these pharmacological manipulations can be quantitatively assessed by measuring their impact on variables in each of these classes. These deviations are easily and safely produced by several classes of drugs which directly or indirectly modify the function of central muscarinic cholinergic networks. Methods for inducing these changes in cholinergic systems and their application to clinical and basic research in the field of affective disorders are highlighted. PMID- 3044960 TI - GAO: volume of federal and state debt skyrockets. PMID- 3044961 TI - How hospitals sabotage PR efforts. PMID- 3044962 TI - Experts: learn contracting before it's too late. PMID- 3044963 TI - HCFA proposes 'puzzling' rule regarding PPS. PMID- 3044964 TI - Hospitals choose to accept HHS malpractice offer. PMID- 3044965 TI - Physician and HCFA roles grow in home health. PMID- 3044966 TI - Catastrophic coverage needs dose of ingenuity. PMID- 3044967 TI - Congress skeptical of warnings about low margins. PMID- 3044968 TI - Patient referrals likely to become a risky business. PMID- 3044969 TI - Cash crisis: a hospital survives margin woes. PMID- 3044970 TI - Feds strive to be monopsonists. PMID- 3044971 TI - Medicare PPO tests costs, utilization control techniques. PMID- 3044972 TI - Citing too much profit, IME slated for cuts. PMID- 3044973 TI - Catastrophic care: more than meets the eye. PMID- 3044974 TI - Immunoprecipitation of a male-specific polypeptide after in vitro translation of testicular poly(A)+ RNA. AB - Poly(A)+ RNAs from male and female gonads of 20-day-old rats were translated in vitro using rabbit reticulocyte lysates. The resulting polypeptides were incubated with specific anti-male (anti-H-Y) antisera raised in female rats by intrasplenic immunization with syngeneic male skin. Using a second antibody, a male-specific polypeptide of molecular weight approximately 18,000 was immunoprecipitated. This male-specific polypeptide was not cotranslationally modified in vitro and appeared to be identical to serological H-Y antigen. PMID- 3044975 TI - The distribution of sex-specific (H-Y) antigens within the seminiferous tubules of the testis: an immunohistochemical study. AB - The cellular distribution of H-Y antigen within the seminiferous tubules of testes from both 20-day-old and adult rats has been examined immunohistochemically. Large amounts of diffuse-staining material surrounding the germ cells were observed within the tubules of 20-day-old rats while the germ cells appeared to have little H-Y positive material on them. In the sexually mature rat, the seminiferous tubules contained cells at various stages of development. Peroxidase staining was evident on many, but not all of these cells. On spermatids and spermatozoa with cytoplasmic droplets attached, peroxidase staining appeared to be present in only a proportion of these cells. PMID- 3044976 TI - Premarket evaluation of Monofluor reagent for detecting Chlamydia trachomatis in adolescent outpatients. AB - A new direct fluorescent antibody reagent, Monofluor, was evaluated for detecting Chlamydia trachomatis in fresh specimens. Monofluor was compared with Micro Trak and with cultivation in McCoy cells. Both direct systems were slightly less sensitive than culture, but no significant differences in specificity or sensitivity were noted between culture, Monofluor, or Micro Trak results. PMID- 3044977 TI - A 24 hour plastic envelope method for isolating and identifying Gardnerella vaginalis (PEM-GVA) AB - A new plastic envelope culture test that is selective for Gardnerella vaginalis was compared with a conventional method. Vaginal specimens from 92 women were cultured. Results from both methods were compared with the results of pelvic examinations and clinic screening tests used to diagnose bacterial vaginosis (BV). G vaginalis was isolated more often in the envelope than by the conventional method from patients with BV and those without, though the difference was not significant. Isolation and identification of G vaginalis was completed in 18-24 hours by the envelope method; the conventional method took a mean of 72 hours (range two to six days). Polymorphonuclear leucocytes (PMNLs) occurred significantly more in specimens from the patients without BV than from those with BV. Both clue cells and a positive amine test reaction were found significantly more in specimens from patients with BV than from those without BV. Clue cells and G vaginalis isolation correlated best with BV (in 47 women), followed by clue cells and positive amine test results (in 39). Adherence of G vaginalis in the envelope also correlated more with BV, clue cells, and positive amine test results (32) than with patients without BV (14). When there were no clue cells and amine test results were negative the results correlated totally with a prediction of no BV. The use of the rapid envelope culture test would have confirmed BV in 20% of the cases where clue cell and amine test results were discordant. PMID- 3044978 TI - Treating chancroid with enoxacin. AB - Increasing resistance of Haemophilus ducreyi to antimicrobials necessitates further trials of new antimicrobial agents for treating chancroid. Enoxacin has excellent in vitro activity against H ducreyi, and a randomised clinical trial of three doses of enoxacin 400 mg at intervals of 12 hours compared with a single dose of trimethoprim/sulphametrole (TMP/SMT) 640/3200 mg was therefore conducted. Of 169 men enrolled in the study, 86 received enoxacin and 83 received TMP/SMT. Ulcers were improved or cured in 65/73 men treated with enoxacin and 57/70 men treated with TMP/SMT. This difference was not significant. At 72 hours after treatment, H ducreyi was eradicated from ulcers of 72/77 men treated with enoxacin and of 67/74 of those treated with TMP/SMT. Patients with buboes responded equally well to both treatments. Of 100 H ducreyi strains tested, all were susceptible to both 0.25 mg/l enoxacin and the combination of 0.25 mg/l TMP and 5 mg/l SMT. Although most men treated with either regimen were cured, neither regimen appeared to be the optimum treatment for chancroid. This study shows the efficacy of enoxacin for a soft tissue infection caused by Gram negative organisms. PMID- 3044979 TI - How reliable is cell culture for detecting Chlamydia trachomatis in patients with urogenital inflammation? PMID- 3044980 TI - Radiotherapy for the treatment of pulmonary complications of paraquat poisoning. AB - The effect of radiotherapy on the pulmonary damage caused by paraquat (24% solution of 1,1'-dimethyl-4,4'-bipyridylium dichloride) was investigated in a preliminary series of nine patients. Paraquat intoxication was diagnosed by quantitative analysis of urine and plasma using colorimetry after extraction of a cation exchange column. The irradiation was given as a planned procedure from day 2 in cases 1 to 7, and after changes in chest X-ray were recognized in cases 8 and 9. A cobalt-60 unit with opposed anterior and posterior portals was used to give a total dose of 12.50 Gy (uncorrected) over 10 fractions, sparing the pericardium and mediastinum as much as possible. Each fraction consisted of 1.25 Gy (125 rad) given once a day, alternating between the left and right lungs. Radiological diagnosis consisted of clear chest X-rays (cases 2 to 4), pulmonary oedema (cases 7 and 9) and interstitial infiltrates (cases 1, 5, 6 and 8). Cases 1, 5 and 8 survived. Case 8 had residual interstitial infiltrates three months after ingestion but these had cleared one month later, suggesting that the diagnosis of irreversible fibrosis should only be made after a follow-up period of at least one year. The results have failed to show a definite benefit, but do support the fact that radiotherapy should be assessed carefully in a randomized trial which is now in progress. PMID- 3044981 TI - Inability of adult circulating haemopoietic stem cells to sustain haemopoiesis in mouse fetal liver microenvironment. AB - Liver rudiments were removed from mouse embryos at Day 9 of gestation, before the 28-somite stage, when they are still not colonized by extrinsic haemopoietic stem cells (HSC), and they were grafted under the kidney capsule of syngeneic adult mice. Hepatocytes differentiated normally, but no colonization by HSC occurred. As control, we used precolonized thymic rudiments taken at Day 10 of gestation, when they are still included in the third branchial arches. They were grafted in the same conditions as liver rudiments and they became colonized by extrinsic HSC that gave rise to lymphocytes. So, adult HSC are not able to colonize the fetal liver rudiment while they have the capacity to home the fetal thymus. We discuss these results in terms of an ontogenic maturation of HSC that could change their homing capacities around birth. PMID- 3044982 TI - Influence of tuftsin-like synthetic peptides derived from C-reactive protein (CRP) on platelet behaviour. AB - C-reactive protein (CRP) is an acute-phase reactant that modifies platelet function differently, depending upon its physiochemical state. Aggregated and ligand-complexed forms of CRP initiate the activation of platelets, whereas naturally occurring CRP peptides inhibit platelet activation. The present study documents neutral proteases of the polymorphonuclear leucocyte (PMN) to cleave CRP into reaction products with the potential to inhibit platelet activation, and explore the structure-function relationships involved in the regulation of platelet activation by CRP using synthetic CRP peptides. Evidence was obtained that (i) a minimum of two linear functional domains exist within CRP that influence platelet activation; (ii) they reside in the mid-portion and at the C terminus of the CRP molecule; (iii) the mid-portion domain inhibits platelet activation stimulated by adenosine diphosphate (ADP) or acid-soluble collagen, whereas the C-terminal domain initiates platelet activation; (iv) the functional expression of the C-terminal domain is maximized when the linear peptide is immobilized on latex; and (v) both CRP domains contain a homologue of the immunoregulatory signal peptide, tuftsin. These data suggest that the molecular mechanisms by which platelet processes are modulated by CRP may be related to the presence of tuftsin homologues in CRP. PMID- 3044984 TI - [Oculocutaneous syndrome following total biliopancreatic diversion]. PMID- 3044985 TI - [Evaluation of a new immunoenzyme method for demonstrating Treponema-specific IgM in human syphilis]. PMID- 3044983 TI - Assessment in mice of a synthetic peptide-based vaccine against the sporozoite stage of the human malaria parasite, P. falciparum. AB - The anti-P. falciparum sporozoite vaccine consisting of the synthetic peptide, Ac Cys-(NANP)3, conjugated to the protein tetanus toxoid (TT), [Ac-Cys-(NANP)3]25 TT, is currently undergoing human trials. The purpose of the present study was to assess various immunological parameters of this vaccine in mice, which have practical implications in humans. Two injections of [Ac-Cys-(NANP)3]25-TT adsorbed to Al(OH)3 were required to elicit a high antibody response against both Ac-Cys-(NANP)3 and TT. The vaccine initiated equivalent Ac-Cys-(NANP)3 priming for a secondary IgG response in 1-week-old and adult mice. Immunization of female mice with TT or [Ac-Cys-(NANP)3]23-TT prior to mating resulted in offspring that passively received anti-Ac-Cys-(NANP)3 and/or anti-TT antibody and that had reduced secondary responses to Ac-Cys-(NANP)3 and TT. Tertiary challenge with vaccine could substantially overcome such inhibition. Preimmunization of adult mice with TT resulted in a specific inhibition of the anti-Ac-Cys-(NANP)3 antibody response that disappeared following tertiary challenge with the vaccine. The conjugate initiated an antibody response against Ac-Cys-(NANP)3 and TT in mice of 16 different genotypes; only very low T-cell proliferative responses to (NANP)3 were observed for some of these strains. Mice injected with (NANP)3 coupled to protein demonstrated a secondary response to Ac-Cys-(NANP)3 when challenged with (NANP)3 on a heterologous carrier, indicating that B-cell priming alone may be sufficient for a secondary antibody response. These results demonstrate that the vaccine has favourable and unfavourable characteristics in mice; the potential for both exists in humans. PMID- 3044987 TI - [Minocycline in the therapy of Chlamydiaceae and Neisseria gonorrhoeae infections (diagnosed by immunoenzyme technics)]. PMID- 3044988 TI - [The hypereosinophilia syndrome]. PMID- 3044989 TI - Bone marrow transplantation in the treatment of severe immunodeficiencies: possibilities and problems. AB - Infants and children suffering from severe primary immunodeficiencies may be cured by bone marrow transplantation from a healthy donor. Data obtained in 14 European centers show that about 60% of the patients are surviving disease-free, if they were grafted with bone marrow cells from an HLA-identical related donor. Results of transplantation of T-cell depleted bone marrow from an HLA haploidentical related donor were also excellent in infants with severe combined immunodeficiency, with 60% recovery. This therapy is superior to transplantation of fetal tissues. HLA-haploidentical T-cell depleted marrow transplantation for other severe immunodeficiencies was less successful. This was mainly due to failure of engraftment, despite intensive conditioning of the recipient, and to infectious complications e.g. by reactivation of latently present viruses. PMID- 3044986 TI - [The personal computer in dermatology. Use in filing, diagnosis and research]. PMID- 3044990 TI - Immunodeficiency due to defects of polymorphonuclear leukocyte function. AB - Functional defects of polymorphonuclear leukocytes can lead to clinical immunodeficiency. Understanding these conditions requires a knowledge of normal polymorph physiology and the means to analyse various aspects of polymorph function. Primary diseases of polymorph function are rare, whereas disorders secondary to other diseases are more common. Prompt diagnosis of these conditions leads to better patient management. PMID- 3044991 TI - The Sda antigen in the human kidney and colon. AB - The range of concentration of the Sda blood group antigen has been determined in the human adult kidney and colon. No Sda antigen was found in 2% of kidneys. Immunofluorescent studies of the kidney showed that Sda is present in the distal convoluted tubules and collecting ducts and occurs in the same location as Tamm Horsfall protein. No Sda antigen was detected in about 2% of colons. In an additional 6%, no antigen was detected in saline extracts but was present in an insoluble form. In the colon, Sda is sited on the brush borders of the epithelial cells and in the goblet cells. No Tamm-Horsfall protein was identified in the colon. PMID- 3044992 TI - Trends in cancer incidence in Singapore 1968-1982. PMID- 3044993 TI - Adenosine in renin-dependent renovascular hypertension. AB - Our previous studies support the hypothesis that activation of the renin angiotensin system by renal ischemia elevates adenosine levels and that adenosine acts in a negative feedback loop to limit renin release and to mitigate some of the hypertension-producing effects of angiotensin II. To further test this hypothesis, we compared the time course of caffeine-induced increases in plasma renin activity with the time course of changes in plasma levels of adenosine in two models of renin-dependent renovascular hypertension. Also, we compared the effects of caffeine on plasma renin activity and arterial blood pressure in renin dependent versus renin-independent renovascular hypertension. In comparison to sham-operated rats, plasma levels of adenosine in the left and right renal veins and aorta were elevated severalfold in two-kidney, one clip rats (2K1C) 1 week after left renal artery clipping. However, adenosine levels declined during the second and third weeks after clipping. In 2K1C rats treated chronically with caffeine, plasma renin activity was markedly elevated during the first week after operation as compared to non-caffeine-treated 2K1C rats. However, during the second and third weeks after clipping, caffeine had lesser effects on plasma renin activity. A temporal relationship between plasma adenosine levels and caffeine-induced hyperreninemia was also observed in rats with aortic ligation. Caffeine accelerated hypertension in 2K1C rats and rats with aortic ligation (renin-dependent renovascular hypertension), but it had no effect on plasma renin activity or blood pressure in one-kidney, one clip rats (renin-independent renovascular hypertension). These results lend further support to the hypothesis that adenosine functions to mitigate the renin-angiotensin system in renin dependent renovascular hypertension. PMID- 3044994 TI - Cerebral circulation in chronic arterial hypertension. AB - Several new concepts have emerged recently regarding the effects of chronic hypertension on cerebral blood vessels. First, hypertrophy of large cerebral arteries in chronic hypertension attenuates increases in pressure of downstream vessels and protects the cerebral microvasculature. Second, in contrast to large cerebral arteries, which become less distensible during chronic hypertension, distensibility of cerebral arterioles increases during chronic hypertension despite hypertrophy of the arteriolar wall. Third, dilatation of cerebral blood vessels with disruption of the blood-brain barrier, and not vasospasm, appears to be the critical factor in the pathogenesis of hypertensive encephalopathy. This concept is supported by the finding that cerebral edema in stroke-prone spontaneously hypertensive rats is preceded by vasodilatation and disruption of the barrier. Fourth, alterations of endothelium-mediated dilatation may impair vasodilator responses in chronic hypertension and predispose to ischemia. Finally, chronic hypertension impairs dilatation of collateral blood vessels in the cerebral circulation. The implication of this finding is that increased susceptibility to cerebral infarction in chronic hypertension may be related in part to compromised responses of the collateral circulation. PMID- 3044995 TI - Roles of motility, chemotaxis, and penetration through and growth in intestinal mucus in the ability of an avirulent strain of Salmonella typhimurium to colonize the large intestine of streptomycin-treated mice. AB - Previously, it had been shown that an avirulent strain of Salmonella typhimurium, SL5316, with wild-type lipopolysaccharide (LPS) was a far better colonizer of the streptomycin-treated CD-1 mouse large intestine, was far more motile, did not bind to mouse intestinal mucus nearly as well as, but penetrated through a layer of intestinal mucus in vitro far better than an almost isogenic LPS-deficient transductant, SL5325. In the present investigation, a nonflagellated transductant, SL5681, and a nonchemotactic transductant, SL5784, were isolated from SL5316 and tested for relative colonizing ability versus SL5316 (smooth) and SL5325 (rough) in streptomycin-treated mice. In addition, the Salmonella strains were tested for their ability to grow together in cecal intestinal mucus and in cecal luminal contents, for their tumbling and swimming activities after growth in cecal mucus, and for their ability to adhere to and travel through cecal mucus in vitro. The data show that the nonflagellated and nonchemotactic derivatives colonized large intestine nearly as well as their parent and were far better colonizers than the LPS-deficient mutant, that all the strains grew equally well in cecal mucus but did not grow in cecal luminal contents, and that cecal mucus grown strains lost tumbling and swimming activities. Furthermore, the LPS deficient strain adhered to cecal mucus far better but penetrated mucus far worse than did the nonflagellated transductant, the nonchemotactic transductant, and the parent. Thus, motility and chemotaxis do not appear to play a major role in the ability of the avirulent S. typhimurium strains to colonize the mouse large intestine, colonization may require growth in cecal mucus but does not depend on growth in cecal luminal contents, growth in cecal mucus inhibits S. typhimurium motility, and increased adhesion of the LPS-deficient mutant to cecal mucus and its poor ability to penetrate cecal mucus may play a role in its poor intestine colonizing ability. PMID- 3044996 TI - Use of lymphokines in treatment of experimental intra-abdominal abscess caused by Bacteroides fragilis. AB - The role of cell-free soluble factors (lymphokines) derived from mitogen activated splenic cells of mice previously immunized against Bacteroides fragilis was evaluated in the treatment of B. fragilis intra-abdominal abscess (IAA). Neither clindamycin nor lymphokines alone were effective against an established B. fragilis IAA, but the combination of clindamycin and lymphokines decreased the abscess size and bacterial counts in the majority of animals. This suggests that the synergy of lymphokines with clindamycin effects cure of IAA caused by B. fragilis and that lymphokines might have an application as adjuncts to conventional antimicrobial therapy in this setting. PMID- 3044998 TI - Role of H-2 and non-H-2 genes in control of bacterial clearance from the spleen in Salmonella typhimurium-infected mice. AB - The ability of mice to clear Salmonella typhimurium from their spleens in the late phase of infection was studied after inoculation with a temperature sensitive mutant. Clearance of bacteria was delayed in C57BL/6 mice compared with BALB/c, C3H/HeJ, DBA/2, A/J, and CBA mice. The responses of F1 hybrids, backcrosses, and recombinant inbred strains derived from C57BL/6 and BALB/c (both Itys) and of H-2 congenic mice were analyzed. The results showed that the low rate of bacterial clearance was recessive, that the rate of clearance was under polygenic control, and that an H-2-linked gene(s) plays a major role. Among H-2 congenic mice with a C57BL/10 background, three phenotypes of bacterial clearance could be distinguished: high (H-2j, H-2q, and H-2u), intermediate (H-2d, H-2f, H 2k, H-2p, H-2r, H-2s, and H-2v), and low (H-2b) rates. The effect of the H-2 complex was apparent with different genetic backgrounds (Itys and Ityr). In recombinant inbred strains derived from C57BL/6 (Itys) and A/J (Ityr) mice, the effect of the H-2b haplotype on bacterial clearance appeared to be fully expressed only in strains carrying the Itys allele. PMID- 3044997 TI - Direct cytotoxic action of Shiga toxin on human vascular endothelial cells. AB - To help explain a role of the Shiga toxin family in hemorrhagic colitis and hemolytic-uremic syndrome in humans, it has been hypothesized that these toxins cause direct damage to the vascular endothelium. We now report that Shiga toxin purified from Shigella dysenteriae 1 does indeed have a direct cytotoxic effect on vascular endothelial cells in cultures. Human umbilical vein endothelial cells (HUVEC) in confluent monolayers were reduced 50% by 10(-8) M Shiga toxin after a lag period of 48 to 96 h. In comparison, nonconfluent HUVEC were reduced 50% by 10(-10) M Shiga toxin within a 24-h period. These data suggest that dividing endothelial cells are more sensitive to Shiga toxin than are quiescent cells in confluent monolayers. Both confluent and nonconfluent HUVEC specifically bound 125I-Shiga toxin. However, in response to the toxin, rates of incorporation of [3H]leucine into protein were more severely reduced in nonconfluent cells than in confluent cells. Toxin inhibition of protein synthesis preceded detachment of cells from the substratum. The specific binding of 125I-Shiga toxin to human endothelial cells and the cytotoxic response were both toxin dose dependent and neutralized by anti-Shiga toxin antibody. Heat-denatured Shiga toxin was without the cytotoxic effect. In addition, the complete culture system contained less than 0.1 ng of bacterial endotoxin per ml, as measured by the Limulus amoebocyte lysate test. PMID- 3045000 TI - Inhibition of attachment of Escherichia coli RDEC-1 to intestinal microvillus membranes by rabbit ileal mucus and mucin in vitro. AB - Intestinal mucus is postulated to play a role in preventing colonization of the gastrointestinal tract by microbial pathogens. To evaluate the ability of both crude mucus and purified mucin, a glycoprotein of goblet cell origin, to inhibit mucosal adherence of enteric pathogens, we examined whether mucus and mucin derived from rabbit ileum interact with the rabbit enteropathogen Escherichia coli RDEC-1. We examined the manner in which mucus and mucin inhibited adherence of bacteria to rabbit ileal microvillus membranes (MVMs) in vitro. The purity of the mucin preparation was demonstrated by polyacrylamide gel electrophoresis before and after reduction and by showing that an antiserum raised to the mucin localized to goblet cells in rabbit intestine. Using radioactive labeling of bacteria, we quantitated attachment of RDEC-1 to MVMs, mucus, and mucin that had been immobilized on polystyrene microtiter wells. Binding of RDEC-1 to MVMs was also determined after preincubation of organisms with crude ileal mucus and purified mucin. RDEC-1 bound to both crude mucus and purified mucin when they expressed lectinlike adhesions, previously designated attachment factor rabbit 1 pili. Adherence of piliated RDEC-1 to MVMs, mucus, and mucin was significantly greater than when the bacteria were nonpiliated. Binding of piliated RDEC-1 to MVMs was decreased by preincubation of bacteria with both crude mucus (45.6 +/- 4.2% of control) and purified mucin (50.2 +/- 5.8%). These data indicate that the E. coli enteropathogen RDEC-1 can bind to purified glycoproteins of goblet cell origin and that adherence of these bacteria to mucin is mediated by expression of pili. The findings also support a role for intestinal mucus and its principal organic constituent, mucin, in preventing adherence of a known E. coli enteric pathogen to apical MVMs of enterocytes. PMID- 3044999 TI - Iron regulation of the cloned diphtheria toxin promoter in Escherichia coli. AB - Regulation of the diphtheria toxin promoter by iron was studied in Escherichia coli by using a galK transcriptional fusion. A fragment of the toxin (tox) operon containing the regulatory region was cloned from corynephage beta into a galK transcription vector such that expression of galK activity was controlled by the tox promoter. When E. coli N100 (a galK mutant) harboring this tox-galK fusion plasmid was grown in Luria broth, the specific activity of galactokinase remained constant throughout the exponential phase of growth. When bacteria were shifted from such high-iron medium into low-iron Luria broth, the specific activity of galactokinase increased rapidly, but induction of galactokinase was prevented by the addition of iron to the medium. Measurement of tox-specific mRNA by dot blot hybridization showed that this regulation occurred at the level of transcription. When the plasmid containing the tox-galK fusion was introduced into a fur mutant of E. coli, expression of galK was maximal in both high-iron and low-iron media; but repressibility of galK by iron in this strain was restored by complementation with the fur+ allele. The tox promoter has significant homology with the consensus sequence for other iron-regulated promoters of E. coli that are controlled by fur. These data indicate that the product of the fur gene can function in E. coli as an iron-dependent repressor for the tox promoter from corynephage beta. PMID- 3045001 TI - Genetic relationships among pathogenic strains of avian Escherichia coli. AB - Genetic relationships among 79 strains of Escherichia coli, isolated mostly from diseased chickens, were estimated on the basis of allelic variation at 15 enzyme encoding loci, determined by multilocus enzyme electrophoresis. All 15 loci were polymorphic, with an average of 4.1 allelic states per locus. Comparisons of the observed combinations of alleles among strains revealed 37 distinct multilocus genotypes that were used to define naturally occurring cell lineages or clones. Two-thirds of the isolates were classified into 10 clones, including a single multilocus genotype that accounted for about a third of all isolates. For isolates of these clones, there was a high concordance (76%) between identity in multilocus genotype, O:K:H serotype, and pattern of resistance to five antibiotics. Cluster analysis disclosed two major complexes of closely related clones, in which more than 50% of the isolates were associated with localized infections (airsacculitis and pericarditis). Both complexes contained isolates with serotype O2:K1, indicating that this serotype can occur on diverse chromosomal backgrounds. The results suggest that colibacillosis within avian populations is caused by a relatively limited number of pathogenic clones representing at least two distinct clone complexes. PMID- 3045002 TI - Heterogeneity of hemolytic efficiency and isoelectric point of streptolysin O. AB - Using thin-layer agarose gel isoelectric focusing overlaid with thin-layer erythrocyte agar plates, we found that crude streptolysin O (SLO) consisted of a variety of hemolytic components with different isoelectric points (pIs) and that the distribution of pIs in crude SLO was different even in samples which were produced from a single strain of Streptococcus pyogenes under similar conditions. All of the hemolytic components in crude SLO were shown to have the properties of SLO with respect to their susceptibility to oxygen and anti-SLO serum and their molecular weight. The SLO components showed a single molecular weight of 64,000, but they exhibited various pIs ranging from pH 5.4 to 8.3, with major components showing a pI of 6 and/or 7.5. Further examination revealed the slope of the hemolytic titration curve to be dissimilar among the samples of crude SLO. Since the slope of the hemolytic titration curve of a component appears to be based on its hemolytic efficiency, the value of the slope was designated its hemolytic efficiency index. When SLO was purified by isoelectric focusing, the pI of the components was correlated with its hemolytic efficiency index; hemolytic components with lower pIs exhibited a lower hemolytic efficiency index. These results indicate that SLO consists of heterogeneous components with different pIs and suggest that the differences in hemolytic efficiency indices of SLO components are due to the different electrical charges of SLO molecules, which are related to their polymerization and affect hemolytic efficiency. PMID- 3045004 TI - Effect of intravenous silica on the course of Nocardia asteroides pneumonia. AB - Silica, a known toxin of mononuclear phagocytes, was administered intravenously to mice during Nocardia asteroides pneumonia. Mice that received silica had a sevenfold decrease in the number of peripheral blood monocytes and developed more severe N. asteroides pneumonia than control mice. Lung histology in mice that received silica resembled that of mice with impaired cell-mediated immunity. These results are most consistent with the explanation that silica injures blood monocytes and impairs their contributions to pulmonary host defense. PMID- 3045003 TI - Isolation of Shiga toxin-resistant Vero cells and their use for easy identification of the toxin. AB - Shiga toxin-resistant Vero cells were isolated by treatment of the cells with nitrosoguanidine. These mutant cells were not affected by Shiga toxin at more than 1 microgram/ml, although the parent Vero cells were sensitive to 25 pg of the toxin per ml. Immunofluorescence studies showed that all the mutant cells had lost toxin-binding capacity. The cytotoxic activities of various bacterial cultures against the parent and mutant cells were compared. All samples from 10 strains of Shigella dysenteriae type 1 and all three strains of Escherichia coli O157:H7 tested showed cytotoxicity to the parent cells but not to the mutant cells. Samples from other organisms, such as Shigella flexneri, Shigella sonnei, Clostridium difficile, Aeromonas hydrophila, Aeromonas sobria, and other E. coli strains, either had no effect or were cytotoxic on both the parent and mutant cells. Thus, these mutant cells could be used to identify Shiga-like toxin and distinguish it from other cytotoxins. The results also suggest the presence of a receptor for Shiga-like toxin on Vero cells that is essential for expression of the cytotoxicity of Shiga toxin but is not essential for growth of Vero cells. PMID- 3045005 TI - Nucleotide sequence of streptococcal pyrogenic exotoxin type C. AB - The nucleotide sequence of the gene speC, encoding streptococcal pyrogenic exotoxin type C (SPE C), was determined. The gene encoded a mature protein of 208 amino acids, with a calculated molecular weight of 24,354. The mature amino acid sequence of SPE C was analyzed for homology with the amino acid sequences of streptococcal pyrogenic exotoxin type A, the staphylococcal enterotoxins, and toxic shock syndrome toxin-1. Of these, SPE C shared the greatest amount of homology with streptococcal exotoxin type A. PMID- 3045008 TI - Bonding to tooth structure: clinical and biological considerations. AB - Marginal leakage of tooth restorations is a problem well known to dental practitioners and researchers. The development of agents that provide strong and stable adhesive bonds to both dentine and enamel in the oral environment is a challenge to scientists. The critical area in resin restorations is at the gingival margin where the resin is in apposition to dentine and/or cementum. Therefore, investigations into dentine bonding have been undertaken over the past 20 years. The evaluation of bonding agents includes studies in vitro to investigate their ability to aid the control of microleakage. The biological safety of bonding agents is also very important. Pulpal response to these agents is useful in biological evaluation. Animal studies on the pulpal response of available bonding agents have been reported recently. Precise evaluation criteria have been employed on a limited basis. Long-term clinical studies are now necessary to evaluate the stability of the commercially available bonding agents that have been histologically and biologically evaluated. PMID- 3045006 TI - Dynamic expression of cell surface hydrophobicity during initial yeast cell growth and before germ tube formation of Candida albicans. AB - Expression of cell surface hydrophobicity (CSH) during initial growth of Candida albicans was monitored. CSH of hydrophobic and hydrophilic yeast cells changed within 30 min upon subculture into fresh medium. Morphologic evidence of germination was preceded by expression of CSH. These results indicate that CSH expression is important in C. albicans growth. PMID- 3045009 TI - The use and abuse of aesthetic materials in posterior teeth. AB - This paper summarizes the clinically significant concerns surrounding the use of aesthetic restoratives in posterior teeth. Composite resins are far from ideal and possess some properties that should be of real concern to the dentist. They are not amalgam substitutes. On the other hand, they do have a place in the conservative cavity where aesthetics are desired. The suggested use of composite and porcelain inlays or onlays is an attempt to circumvent some of the concerns generated by the routine use of composites. However, the relative scarcity of clinical studies suggests a limited usage at this time. PMID- 3045007 TI - Asbestos exposure, pleural plaques and the risk of lung cancer. AB - Studies which have evaluated the relationship between pleural plaques and smoking have found a higher prevalence of smokers among persons with pleural plaques. Pleural plaques are a relatively frequent finding among persons with occupational exposure to asbestos. Some studies, but not others, have shown that persons with pleural plaques have a higher risk of lung cancer. None of these studies controlled for the effects of smoking, and since smoking is more prevalent among persons with pleural plaques, it is unlikely that the increased risk of lung cancer to persons with pleural plaques, found in some studies, is due to the pleural plaques. PMID- 3045010 TI - Modern restorative dentistry: a new approach. AB - An analysis of the role of restorative dentistry within oral health care shows that it has broadened its scope over its long history. It started with the replacement of lost teeth and, later, lesions and structural defects of tooth tissue were treated. Progress has been achieved through the development of new materials and procedures, specifically those that ensure microscopic and/or chemical adhesion and those that allow for the removal of infection tissue and its restoration, while protecting the remaining healthy structure of the tooth. Teeth can also be protected even before the initiation of an infected lesion by sealing morphologic features such as pits and fissures. All of these procedures are components of modern restorative dentistry which now contributes to the disappearance of the totally edentulous patient. Dentistry has the responsibility of helping in the generation of sufficient social and economic resource to make this modern restorative dentistry available to the community as a whole. PMID- 3045011 TI - Bonding of restorative resins to enamel. AB - The bond between restorative resin and enamel may be likened to a chain with a series of links. The potentially weakest link is the interface between the resin and the tissue. This link is established by the clinician and is considered to be technique sensitive. An understanding of the steps in clinical method provide a rationale and an aim for each step. A thorough dental prophylaxis to remove deposits, including calculus from enamel, is important in allowing the acidic conditioning agent, namely phosphoric acid, to exert its optimal effect. Conditioning of the enamel raises its surface energy, creates increased surface area and enhances tissue porosity in the outermost enamel. These conditions are achieved through careful isolation of the teeth and thorough washing to remove the acid and reaction products. A dry field is also of paramount importance in obtaining the micromechanical retention of the resin within the enhanced tissue micropores. The polymerized resin within the tissue serves to retain the bulk of resin located upon the enamel surface. The method is a clinically conservative, non-traumatic, biocompatible means by which resins, polymerizable under oral conditions, can be bonded durably to enamel. PMID- 3045012 TI - Bonding of restorative resins to dentine: status of dentine adhesives and impact on cavity design and filling techniques. AB - In conjunction with the acid etch technique for bonding of restorative resins to enamel, an effective bonding to dentine would eliminate the need for retentive undercuts and ensure a tight marginal seal. Bonding to dentine can be achieved through dentine adhesives. These may be divided into Ca2+-bonding and collagen bonding types. Several Ca2+-bonding adhesives are based on phosphate methacrylates. Adhesives of this type mediate a bond strength to dentine not exceeding 10 MPa. In contrast, Bowen's bonding system and the Gluma system yield bond strengths higher than 10 MPa. The two latter systems have recently been conceived in simplified versions. In general, the higher the bond strength to dentine mediated by an adhesive, the smaller are the marginal gaps formed by a composite resin polymerizing in a dentine cavity treated by the adhesive. The size and the shape of the cavity influence the width of the marginal gaps. A cavity having a V-shape gives rise to smaller gaps than box-shaped cavities. A filling technique with two 'inclining' layers reduces the width of marginal gaps. Combination of certain Ca2+-bonding and collagen-bonding adhesives results in increased bond strength and smaller marginal gaps. PMID- 3045013 TI - Production and characterization of a murine monoclonal antibody to Aspergillus fumigatus antigen having IgG- and IgE-binding activity. AB - By employing hybridoma technology, a monoclonal antibody against Aspergillus fumigatus was produced. This antibody, isotyped as IgM k, reacted with 12 of 16 antigens extracted from 9 different A. fumigatus strains. This antibody also reacted with all 3 Aspergillus flavus antigens studied, but not with Aspergillus niger, Aspergillus terreus, Penicillium notatum or Candida albicans antigens. Western blot analysis indicated that this antibody reacted with two concanavalin A (Con-A) binding bands of the A. fumigatus antigen extract. The specific binding antigens were isolated using monoclonal antibody affinity chromatography. When used in immunoassay this fraction demonstrated strong IgG and IgE antibody binding activities against patient sera. The antibody levels against the purified fraction were significantly higher in patients' sera than in normal controls. The purified fraction demonstrated comparable reactivity with the crude A. fumigatus extract against patient sera, but the former has the added advantage of being pure and standardizable for dependable and reproducible results. PMID- 3045014 TI - Proliferative response of human T lymphocytes to a vaccinal preparation of ribosomes from Streptococcus pyogenes. AB - The in vitro lymphocyte-activating properties of a ribosomal preparation of Streptococcus pyogenes were investigated. The preparation was mitogenic for human lymphocytes with a peak of 3H-thymidine incorporation occurring after 3-5 days of culture. The response was abolished by removal of CD3-positive cells and by alteration of accessory cells by exposure to L-leucine methyl ester. Most of the cells synthesizing DNA at the end of the culture expressed CD4 or CD8 but not CD20 antigens. No immunoglobulin synthesis was demonstrable. Although the same preparation was shown to be a T-independent polyclonal B-cell activator of murine cells, it preferentially triggers T cells in humans. PMID- 3045015 TI - New membranes: the promise of ingenuity. PMID- 3045016 TI - Zinc metabolism in uremia. PMID- 3045018 TI - The in vitro bleeding time while using a stable prostacyclin analogue during hemodialysis. AB - A serious disadvantage of preventing clot formation by using prostacyclin (PGI2) to inhibit thrombocyte function during dialysis is that there exists no rapidly measurable monitoring parameter. The "in vitro bleeding time" (in vitro BT) is a new method for measuring primary hemostasis in vitro. Five chronic dialysis patients each underwent two dialyses: 1) with conventional full heparinization, and 2) with the stable PGI2 analogue CG 4203 and additional "low-dose" heparin. The predialytic in vitro BT is longer than normal values and values 1 h after the end of dialysis. While heparin has no significant effect on the in vitro BT, CG 4203 prolongs it concentration-dependently. Infusing CG 4203 at a rate of 25 ng/kg/min, the in vitro BT is extended beyond the measurable range of 800 microliter during the first approx. 150 min of dialysis. During the next approx. 90 min it steadily decreases. PMID- 3045017 TI - Beta 2-microglobulin, amyloidosis, and dialysis. PMID- 3045019 TI - A basic library of microcomputer programs to obtain immunologically relevant information from protein sequences. AB - A set of simple and interactive BASIC microcomputer programs which may be used to obtain immunologically relevant information from protein sequences is described. Program listings include the computation of structural dissimilarity between two protein sequences (STRUDIS), the predicted secondary structure (CHOUFAS), their expected degree of three-dimensional similarity by correlation of sequence hydrophobicities (HYDCOR) and the location of alpha-helical segments in the three dimensional protein structure (HELOC). In addition, a general program is described for running mean calculations (RUNMEAN). All programs are listed in a BASIC version for Apple IIE. PMID- 3045021 TI - Development of an expert system for preoperative assessment of female urinary incontinence. AB - We developed a simple expert system for preoperative assessment of women complaining of involuntary loss of urine and scheduled to undergo surgery for incontinence. The aim of the system was to use the parameters obtained at urodynamic investigation to arrive at the correct diagnosis. We used an IBM-PC with two disk drives and 256k RAM, and the expert system shell EXSYS, a rule based system with the possibility of assigning probabilities to the different solutions. To write the rules forming the knowledge base we used a two-fold approach: we constructed tree diagrams for each diagnosis and calculated the corresponding predictive values (statistical approach), and we added rules based on our experience (heuristic approach). The expert system has been found reliable in a clinical setting and is useful for teaching purposes. PMID- 3045020 TI - Application of a microcomputer-based system in the analysis of infection data at the emergency units of a large hospital. AB - After three years of retrospective study in four emergency units from a large hospital (2000 beds) and analysis of 6283 positive cultures, a microcomputer database system was built to store information concerning nosocomial infections in order to help the clinical staff from those units to study the incidence of 20 bacterial species and their sensitivity pattern evolutions for 27 antibiotics (from samples in 15 different collecting sites). This system was developed as an alternative to the hospital mainframe computer microbiological reports. It put emphasis on graphical outputs instead of the coded tables generated by the bigger system. This orientation and the possibility of sectorial infection data analysis were responsible for the general acceptance of the microcomputer-based system by the clinical staff. As the first practical results, the system was able to detect a particular increase in the incidence of Staphylococcus aureus in surgical emergency units (up to 21.6% in 1982) as well as the dissemination of the antimicrobial resistance patterns of S. aureus and Klebsiella pneumoniae from the surgical units to the clinical ones. The time evolution behaviour of Pseudomonas aeruginosa, Escherichia coli and other nonfermentative Gram negative bacilli was also studied to complete the analysis of the most pathogenic bacterial species found in our emergency units. PMID- 3045022 TI - On automatic diagnosis of pathological saccadic eye movements. AB - A syntactic technique is described for the recognition of saccadic eye movements to distinguish normal saccades from those distorted by brain stem lesions. A digitalized eye movement signal is transformed into a sequence of symbols. Eye movements are then found from this sequence by using a parser. This recognition method appropriately enlarged could be applied as a classifier of saccades to aid in diagnosis. PMID- 3045023 TI - HLA-based local donor/recipient matching in renal transplantation: a microcomputer program. AB - A microcomputer program was developed for selection of HLA matched recipients in renal transplantation. The program provides an easy way to handle donor/recipient matching within a transplant centre's waiting list. It is based on HLA compatibilities instead of mismatches, and ranks the quality of a DR-match above a B-match and an A-match. To obtain an order of total match quality according to number and type of compatibilities a point count system was developed. The program selects all ABO-compatible recipients with the minimal HLA-match required from the waiting list and ranks them according to match quality. PMID- 3045024 TI - Pharmacokinetics of netilmicin in renal insufficiency and hemodialysis. AB - The pharmacokinetics of netilmicin was studied in 26 patients with varying degrees of renal impairment (11 were dialysis patients) in order to determine the influence of kidney function status on the disposition of the antibiotic. The serum level curves of netilmicin follows a two-compartment open kinetic model. Several relationships between pharmacokinetic parameters and renal function indicators are defined. A clinical useful correlation indicates that the half life is approximately 3 times the serum creatinine concentration, and may be used for adjusting the netilmicin dosage in the treatment of patients with impaired renal function. During hemodialysis the serum half-life decreases approximately 10-fold compared with the interdialysis periods. The percentage of dose extracted by hemodialysis during a single 4 h session is 56.1 +/- 6.65%. The dialysis clearance is 87.28 +/- 28.8 ml/min. PMID- 3045025 TI - Effects of moderate low sodium/high potassium diet on essential hypertension: results of a comparative study. AB - It is generally accepted that a significant restriction in sodium intake can lower blood pressure in hypertensive patients and more recently it has also been suggested that a high potassium intake can exert an antihypertensive effect. We have therefore, conducted a double-blind, randomized, cross-over study to evaluate the antihypertensive efficacy of the combination of a modest dietary sodium restriction and a high potassium intake in hypertensive patients of mild and moderate degrees. During the modest sodium (100 mmol/day)/high potassium (130 mmol/day) diet the blood pressure was significantly reduced (-17/-6 mmHg) when compared to the normal diet (160 mmol Na/day and 80 mmol K/day). The blood pressure reduction did not interfere with hemodynamic and humoral responses to dynamic exercise. The modest reduction in sodium intake and increase in potassium content in the diet was well tolerated by the patients. PMID- 3045026 TI - A pilot evaluation of the effect of defibrotide in patients affected by peripheral arterial occlusive disease. AB - This was a random double-blind study versus placebo designed to assess the effectiveness and tolerability of defibrotide in patients with chronic peripheral vascular disease (PVD), Leriche stage II. A total of 20 male outpatients (mean age 58.8 years) were selected and treated with placebo for 2 weeks (run-in period), and then allocated randomly to treatment with defibrotide (200 mg/day, intramuscular, n = 10) or matched placebo (n = 10) for 90 days. Before intake and at 15, 30, 60, and 90 days of treatment, each patient received a Doppler ultrasound examination of the lower limbs, and the residual perfusion index (Winsor) was calculated at rest and immediately after a symptom-limited treadmill test. Absolute walking distance was measured in addition. Blood samples were taken for routine laboratory tests before intake and at 30 and 90 days of treatment. All patients completed the trial without adverse effects; no meaningful alterations of laboratory test returns were detected. Patients receiving defibrotide showed a significant increase of the Winsor index after exercise at 90 days versus basal (mean 0.47 +/- 0.04 vs. 0.43 +/- 0.05, p less than 0.05), and a parallel increase of walking distance (basal 288 +/- 42.3 m vs. 368 +/- 46.6 at 60 days, p less than 0.05, and 407 +/- 64.3 m at 90 days, p less than 0.01). No improvement was seen in the placebo group. These preliminary results suggest that defibrotide may prove beneficial to patients with PVD; further studies are needed to find the most appropriate dosage regime. PMID- 3045027 TI - Effect of cisapride on the post-cholecystectomy upper gastrointestinal transit time. AB - For the evaluation of postoperative upper gastrointestinal transit, the breath hydrogen test is a convenient method, which is simple but more reliable than the recording of clinical parameters such as bowel sounds. The test was used to study the effect of cisapride on the postoperative mouth-to-caecum transit time. Twenty patients undergoing cholecystectomy were tested pre-operatively and on the first postoperative day after having received either 10 mg of cisapride i.v. or matching placebo under double-blind conditions. The median preoperative transit time in the placebo group was 45 min, and the median postoperative transit time at least 3 h longer. The postoperative transit time after cisapride dosing was still longer than before the operation but was significantly (p = 0.02) reduced as compared with that after placebo administration. PMID- 3045028 TI - Maintenance of potassium balance during long-term diuretic therapy in chronic heart failure patients with thiazide-induced hypokalemia: comparison of potassium supplementation with potassium chloride and potassium-sparing agents, amiloride and triamterene. AB - The relative efficacy of potassium chloride, amiloride and triamterene in maintaining potassium and magnesium balance was evaluated in 23 hypokalemic (S-K less than or equal to 3.5 mmol/l) patients with chronic heart failure receiving diuretic therapy. Amiloride and triamterene were administered in a randomized, crossover manner, followed by potassium chloride in an open manner. During a 5 month treatment with hydrochlorothiazide 50 mg twice/day, potassium chloride 1 g twice/day was not as effective as amiloride 5 mg or triamterene 75 mg twice/day in maintaining serum potassium and magnesium and total-body potassium, while amiloride and triamterene seemed to be equally effective. During all three supplementations, a decrease in serum potassium to a hypokalemic level was observed in some patients. The need for higher doses of potassium chloride, amiloride and triamterene was clearly concentrated to the same patients, and correction was easily reached by increasing the respective doses. PMID- 3045030 TI - Cortical and trabecular osteopenia after immobilization. A quantitative histological study of the rat knee. AB - The aim of this investigation was to examine the effect of short-term immobilization on subchondral cortical and trabecular bone tissue in the rat tibia and to determine whether there was any difference when the knee was immobilized in extension or flexion. Thirty-six male rats were used in this study, and in 18 the knee was fixed in extension and in 18 in flexion. The time of immobilization was for 1, 2 and 3 weeks, with 12 animals in each group and 6 knees in extension and 6 in flexion. The following parameters were measured: (1) the mean thickness of subchondral and both periosteal cortices; (2) cortical porosity; (3) trabecular bone volume; (4) relative osteoid volume, and (5) relative osteoid surface. The mean cortical thickness decreased during the period of immobilization and the cortical porosity significantly increased. The trabecular bone volume was unaltered in the first week, but after 3 weeks it had decreased. The relative osteoid volume decreased significantly during the three weeks, and the relative osteoid surface moderately increased. No relationship was found between the quantitative osteopenic alteration of subchondral bone and the position of immobilization. PMID- 3045029 TI - The bridging of large osteoperiosteal gaps using 'Decalbone'. AB - 'Decalbone' was prepared by partial decalcification of human bones obtained from recently amputated specimens. It was then stored in 80 to 90% ethanol in a domestic refrigerator. The decalbone was used to fill large osteoperiosteal gaps in 25 patients. The commonest lesion was a giant cell tumour (21 cases) and various long bones were affected. There were 6 failures with recurrence of the tumour in 3 and uncontrolled infection in 3. The remaining 19 cases were followed up for from 2 1/2 to 7 years. In 10 the decalbone incorporated well, but further reconstructive procedures were needed in 9. Our studies showed that incorporation began at around 6 to 9 months and was complete at about 2 years in the upper limb and 4 years in the lower limb. There was no clinical evidence of an immune response. PMID- 3045031 TI - Purification and primary structure of ostrich insulin. AB - Insulin has been isolated from ostrich pancreas by a procedure of acid ethanol extraction, adsorption onto SP-Sephadex, gel permeation chromatography and HPLC. The primary structure of the ostrich insulin is identical to that reported for the chicken hormone. The isoelectric point as determined by polyacrylamide gel isoelectric focusing was significantly higher than that of the bovine hormone. PMID- 3045032 TI - Primary structure of equine pituitary prolactin. AB - Equine prolactin was determined to be a single chain protein of 199 amino acid containing two tryptophan and six cysteine residues, as found in other mammalian prolactins. The primary sequence of equine prolactin was obtained by automated Edman analyses of S-carboxymethylated protein and proteolytic fragments of modified protein. Of the known prolactin sequences, equine prolactin shows closest homology with porcine (93%) and fin whale (87-91%) prolactins. Genetic mutations have produced changes in 17 of 199 residues of equine prolactin relative to its putative ancestral precursor. Since equine growth hormone has undergone alterations in 4 of 191 residues relative to this putative precursor protein, these results support the theory that prolactins are evolving at a faster rate than growth hormones. Consistent with the previously determined circular dichroic spectrum of equine prolactin, 60% of the protein is predicted to form alpha helices. PMID- 3045033 TI - The placebo effect during a double blind trial of recombinant alpha 2 interferon in multiple sclerosis patients: immunological and clinical findings. AB - A double-blind, placebo controlled trial of recombinant alpha 2 interferon in relapsing/remitting multiple sclerosis patients was performed to assess the clinical and immunological responses to treatment. This study demonstrated that natural killer (NK) cell activity, known to be enhanced by interferon (IFN) treatment, increased during the first week of treatment in both the IFN and placebo treatment groups. After the first week of treatment NK cell activity returned to baseline levels in both groups, and subsequently declined in the IFN treatment group. Patients in both groups improved clinically, as evidenced by a reduction in the exacerbation rate. Furthermore, a similar incidence of adverse reactions to treatment were reported by both groups. The mechanism underlying the response to placebo is not known. However, it is unlikely that the long term clinical improvement (one year) in either group is related to the transient increase in NK cell activity. PMID- 3045034 TI - Deregulation of hypothalamic opioid and luteinizing hormone releasing factor (LHRF) activity and its relevance to Tourette's syndrome. PMID- 3045035 TI - Historical and philosophical background of neuroimmunomodulation. PMID- 3045037 TI - Neuroplasticity and the progression of Alzheimer's disease. AB - A neurobiological hypothesis is proposed to explain the relation between the percentage cell loss in the cholinergic basal forebrain and the density of neuritic plaques in cortex, as found by Arendt et al. (1985) in Alzheimer's disease: When cells in the cholinergic basal forebrain die, their cortical synaptic target sites can be reoccupied by axonal sprouting of other neurons from the basal forebrain. This neuroplasticity hypothesis leads to equations that are consistent with the quantitative data of Arendt et al. (1985), and it makes specific predictions that can be tested experimentally. Moreover, this hypothesis suggests that the more rapid course of the presenile form of Alzheimer's disease and its more extensive pathology can be understood as a consequence of the decline in neuroplasticity with age. PMID- 3045036 TI - Modulation of thymic endocrine function by thyroid and steroid hormones. AB - The thymic epithelium is responsible for the secretion of thymic hormones that intervene in some steps of intra- and extra-thymic T cell differentiation. In the present paper, we studied the in vivo and in vitro influences of thyroid and steroid hormones on the secretion of thymulin, one of the chemically-defined thymic hormones. Triodothyronine injected in young mice or applied into supernatants of cultured thymic epithelial cells resulted in an increase of thymulin synthesis and secretion, respectively evaluated by the analyses of thymulin containing cells and thymulin levels. The influence of steroids was investigated in vivo by steroid depletion (adrenalectomy and/or castration) and in vitro by the addition of various steroids and/or their specific antagonists to cultured thymic epithelial cells. Surgical ablation of adrenals and/or gonads induced a transient depletion of circulating thymulin and, by feedback mechanism, an increase in the numbers of thymulin containing cells. From the in vitro data we could conclude that the various steroid hormones (that exhibited a stimulatory effect) can act directly on the thymic epithelium, probably via specific receptors. The bulk of data above described represents a strong evidence for the physiological involvement of the neuroendocrine network, on the hormonal function of the thymic epithelium. PMID- 3045038 TI - Neural perspectives of cerebral correlates of giftedness. AB - Giftedness is defined as some special endowment or propensity for creativity, skill, and eminent achievement, found in relatively few individuals among the population. A high order of mental power (IQ), creativity, and motivation (task commitment) appear to be the most universally recognized attributes of the gifted. This report summarizes current knowledge of the cerebral correlates of intelligence and creativity, including physiological measures of EEG, cortical power spectrum, brain evoked potentials, and positron emission tomography. Controversy, debates, contentions, formal hypotheses, and research issues are considered. We are especially interested in the formulation of the deterministic function of EEG-brain dynamics. A CHAOS modeling on hierarchy of cognitive organization and cerebral processing in the gifted is suggested. PMID- 3045039 TI - Nervous control of the phagocytic system. AB - The complexity of the immune response, including numerous functional changes (thermoregulatory, circulatory, respiratory, metabolic etc.) led us (Benetato, Baciu & Vlad, 1945) to hypothesize nervous hypothalamic control of this important homeostatic response. This work synthesizes the experimental results obtained in the last four decades by us on the nervous control of the phagocytic system. Unfortunately at that time those findings were uninteresting, immunity being satisfactorily explained as a purely cellular process. The phagocytic response of the mobile (neutrophils and monocyte) system to i.v. administration of bacteria, may be reproduced by direct electrical stimulation of the hypothalamic tuberomammillary area, and by electroshock. It is blocked after sectioning the spinal cord, or after barbiturates. Using the original "isolated head" technique, applied to dogs, it was demonstrated that the injection of Salmonella typhi murium suspension in the head circulation, may activate phagocytosis in the body, isolated humorally, only through nervous effector ways, which are conserved. The current explanation is that the bacteria in the common circulation of head activates macrophages, producing the endogenous pirogen factor (II 1), which through area hypothalamica anterior acts on the tuberomammillary zone, triggering the phagocytic response by neural effector pathways. Phagocytosis stimulating substances appeared in blood, some as a result of activation of protease systems of the blood (coagulation, fibrinolysis, kininforming and complement). Epinephrine activates peripherally the fibrinolytic and kinin forming systems. Neuroimmunomodulatory effects were also demonstrated for the RES. PMID- 3045040 TI - The basal ganglia and pain. AB - The basal ganglia are associated with motor functions of the brain, although it is becoming clear that they may subserve many nonmotor functions as well. A review of the literature reveals an association between the basal ganglia and pain, and raises the question of a new function for the basal ganglia in selective attention. PMID- 3045041 TI - The epidemiology of alcohol consumption in Spain. AB - This paper is a comprehensive review of the available information about alcohol consumption in Spain. Alcohol use is the most severe drug problem among both young people and adults; almost half the Spanish population drink daily, and more than a fourth drink a quantity of alcohol sufficient to cause health problems in the long term. The consumption of alcohol may be considered to be culturally rooted when compared with other drug use which follows a more recent epidemic pattern. For those who use other drugs, for example cannabis and cocaine, alcohol is commonly used in conjunction. Clearly the level of alcohol use and abuse is high in Spain and constitutes a severe public health and socioeconomic problem. PMID- 3045042 TI - Youth unemployment and health effects. AB - Psychological and sociomedical unemployment research have become an important feature of public concern in the industrialised world. This paper focuses on research into youth unemployment and health effects from an international perspective and investigates more closely the following areas: Unemployment and health: research and the political public Youth unemployment: extent and societal coping Young and unemployed--a special problem? a. Skill utilization and social support b. Psychiatric morbidity c. Vulnerability and age d. "Americanization" of youth The demand for a "social guarantee" PMID- 3045043 TI - Prolactins of pregnancy and their cellular source. PMID- 3045044 TI - The role of hepatocytes and sinusoidal cells in the pathogenesis of viral hepatitis. PMID- 3045045 TI - "Leaky" cells of glandular epithelia. PMID- 3045047 TI - Nutrition and hydration for patients: the constitutional aspects. PMID- 3045046 TI - Membrane oligosaccharides: structure and function during differentiation. AB - Recent results gathered by normal light microscopy, immunocytochemistry, fluorescent-analog cytochemistry, and electron microscopy have allowed an improved interpretation of ameboid movement and related phenomena. 1. The contractile system responsible in Amoeba proteus for the generation of motive force for protoplasmic streaming and a large variety of dynamic activities is represented mainly by a thin cortical filament layer at the cytoplasmic face of the cell membrane (Fig. 18I). During normal locomotion this layer exhibits a distinct structural and physiological polarity with three different zones: a zone of reformation at the front (A), a zone of contraction in the intermediate cell region (B), and a zone of destruction at the uroid (C). 2. Two types of filaments participate in the formation of the cortical layer: (1) randomly distributed thin (actin) filaments exhibiting a parallel orientation in the anterior (Fc1) and a disordered arrangement in the intermediate and posterior cell region (Fc2; see also Fig. 17b), and (2) thick (myosin) filaments in close association with F actin and mostly restricted to the intermediate and posterior cell region (Fc2). 3. The internal hydraulic pressure generated by localized active contraction of the cortical layer is transmitted to the endoplasm via the cell membrane and converted into directed streaming by a gel-sol gradient of decreasing viscosity between the uroid and the front. Calcium ions, ATP, and regulative proteins (profilin and a kinase) play an essential role in controlling both the interaction of actin and myosin and the sol-gel state of the cytoplasmic matrix. 4. Any alterations externally induced in the polarity of the cortical filament system by chemical or physical stimulation and inhibition cause immobilization of the amebas (Fig. 18II) with characteristic changes in (1) cell shape (spherulation and cell flattening), (2) membrane dynamics (cytotic and cytokinetic activities), and (3) cytoplasmic organization (hyalogranuloplasmic separation). pseudopodial tip (Fig. 18III, b----c, d----e), (3) destruction of the old layer at the hyalogranuloplasmic border (Fig. 18III, c,e), and (4) alternate solation (Fig. 18III, b and d) and gelation (Fig. 18III, c and e) of the hyaloplasm between the layer and the plasma membrane. The retraction of pseudopodia is accomplished by a local contraction of the cortical layer in conjunction with a simultaneous gel-sol transformation of the ectoplasmic cylinder. 6. The expression of a rather complex cytoskeleton which is composed not only of microfilaments and associated proteins, but also of intermediate- and microtubularlike structures has to be considered in future PMID- 3045048 TI - Intraoperative ultrasonography in cystic brain lesions. AB - Four neurosurgical patients were operated on under real-time ultrasonographic guidance. Two had cystic tumors which were drained through the unincised dura. In one patient a brain abscess was located intraoperatively by ultrasound after a small craniotomy. In a fourth patient with multiple abscesses, one abscess was drained under ultrasonographic guidance through a previous craniotomy. Using intraoperative ultrasonography the length of the operation can be shortened and normal brain tissue spared. Intraoperative ultrasonography is of special value in cystic lesions which can be drained under ultrasonographic guidance. PMID- 3045050 TI - Clinical experience with a new device for insulin injection. PMID- 3045049 TI - Ceftizoxime vs. cefotaxime--a comparative randomized multicenter study. AB - One hundred and fourteen hospitalized patients with moderate or severe infections were assigned at random, in four medical centers, to receive either ceftizoxime or cefotaxime, administered intravenously in a dosage of 1 to 2 g every 8 h. Of 96 patients evaluable for efficacy, 24 (25%) had bacteremia, 46 (48%) had urinary tract infections and 9 (9%) had pneumonias. Half the patients had been treated ineffectively by other antibiotics prior to the study drug treatment. The overall clinical efficacy was 90% in both treatment groups and 83% in both groups with bacteremia. All patients with urinary tract infection were cured by both agents. Bacteriological eradication rate was 95% in both groups. Adverse reactions, though mild, were more frequent in the cefotaxime group (13.5%) than in the ceftizoxime group (6.8%); superinfection rate was higher in the ceftizoxime group. Both antibiotics were highly and equally efficacious in the therapy of severe infections in hospitalized patients. PMID- 3045051 TI - 15(R)15 methyl prostaglandin E2 (Arbacet) has no effect on human gastric cell labeling index. PMID- 3045052 TI - H.M. Parker lecture. Radiation protection standards: a historical perspective. AB - The historical development of radiation standards limiting exposure to people is given in this paper. Since its purpose was to provide information to the public, the number of units is limited to two, and mathematical expressions are avoided. Emphasis is placed on the role of the National Council on Radiation Protection and Measurements and the International Commission on Radiological Protection rather than on regulations because the recommendations of these groups have played a strong part in the formulation of regulations. PMID- 3045054 TI - The appointed day. PMID- 3045053 TI - Will radiation control be by reason or regulation? AB - Following a very brief review of the development of our present radiation protection philosophy, attention is directed to what the author sees as current problems. The only prognostication will be that at least certain of the problems outstanding or developing today will be among those that will have to be addressed in the coming four decades. For the past six decades, the major effort has been the development of the science and philosophy of radiation biology and its application to protection against radiation hazards, whether real or imagined. For three decades there has been public concern about radiation generally, the core of the problem being inadequate education of the public and the media on the subject. For a little over one decade, there has been rapidly developing growth of tort litigation generally, involving ionizing radiation in particular. These are the three major lines of attention in the radiation protection area today; between two of them there are already some of the aspects of "the arena." Behind all three lies an overwhelming lack of understanding by, and education of, the public, which shows mostly in the public fetish for absolute safety which, of course, cannot be. These must be the major concerns of the radiation-protection community in the coming four decades. PMID- 3045055 TI - [ENT medical findings in obstructive sleep apnea syndromes]. AB - Correlations between the manifestation of obstructive sleep apnoea syndrome (OSAS) and anatomical or functional changes in the upper respiratory tract remain controversial. The correlation between obstruction of the upper respiratory tract and the degree of sleep apnoea syndrome was investigated in 60 patients with obstructive sleep apnoea (diagnosed by polysomnography) and in 55 healthy controls. After clinical examination, rhinomanometry and determination of the size of the lower jaw and oropharynx, the motility of the pharyngeal walls during Mueller's manoeuvre was evaluated by flexible endoscopy. No significant anatomical or functional differences were observed between OSAS patients and healthy controls. There was no correlation between the degree of OSAS (expressed by the apnoea index) and pharyngeal size. Although no specific statement concerning diagnosis or degree of OSAS can be made on the basis of an otolaryngological examination, OSAS patients should always undergo otolaryngological examination to exclude pharyngeal disease. PMID- 3045056 TI - [Mycologic analyses of house dust in patients with mycotic infections of the paranasal sinuses and in healthy probands]. AB - Dust samples from 30 patients suffering from an aspergillus sinusitis and from 20 healthy persons were analysed for moulds as a possible cause of mycotic sinusitis. Questionnaires were also distributed to assess the ecological environment of the patients. Mycological analyses of the dust samples showed no statistical difference in the presence of moulds, especially those of Aspergillus fumigatus and Aspergillus niger. Both moulds were found to roughly the same extent in 75%-95% of subjects. The questionnaires showed that most patients were exposed to moulds during work as well as in their spare time. The case history always showed a chronic sinusitis. Thus it appears that the development of a mycotic sinusitis is caused by many factors. PMID- 3045057 TI - [Herpes zoster cephalicus]. AB - The clinical features of ENT infections induced by the herpes zoster virus are presented. Detailed descriptions of two cases affecting the ear and one affecting the larynx are given, supplemented by photographs. The possible cranial nerve lesions are demonstrated by a review of the literature. An attempt is made to assign the distribution of dermomucosal eruptions and associated sensory and motor dysfunctions to the appropriate neuroanatomical structures. PMID- 3045058 TI - More on hemolytic uremic-like syndrome. PMID- 3045059 TI - Prostatic adenocarcinoma with osseous metastases in a dog. AB - Bone scintigraphy was used to diagnose osseous metastasis of prostatic adenocarcinoma in a 10-year-old dog with neck pain and ataxia and a large, sensitive prostate gland. Although radiography revealed a normal spine, prostatic fluid cytologic and ultrasonographic findings were compatible with prostatitis or neoplasia. Scintigraphic hot spots were seen in the axial skeleton, ribs, pelvis, humerus, and femur and corresponded to sites of metastatic prostatic adenocarcinoma. PMID- 3045060 TI - Current knowledge on paratuberculosis. PMID- 3045061 TI - Biologic and economic risks associated with use of bovine somatotropins. PMID- 3045062 TI - Somatotropin safety--a short regulatory overview. PMID- 3045063 TI - Ways that veterinary medicine can help alleviate hunger in Africa. PMID- 3045064 TI - Economic modeling of the use of gonadotropin-releasing hormone at insemination to improve fertility in dairy cows. AB - Economic and sensitivity analysis methods were used to evaluate financial returns from use of gonadotropin-releasing hormone (GnRH) at the time of insemination to enhance fertility of dairy cows. A computer spread sheet was used to determine the best service(s) for GnRH treatment, the increase in conception rate required for economic benefit from treatment, and how profits from GnRH treatment are affected by drug cost, herd reproductive efficiency, and production costs. Financial returns increased from use of GnRH at insemination under most herd conditions. Herds with conception rates less than or equal to 45% benefited from GnRH treatment at any 1 or 2 inseminations. Herds with conception rates greater than or equal to 60% benefited from GnRH treatment only at second or later services. Selection of second and/or third insemination as the GnRH treatment service usually resulted in the greatest total return. The enhancement of fertility necessary to achieve the break-even point with GnRH treatment at third service was 2% for low- and 5% for high-conception-rate herds. Base-line herd conception rates, estrus detection efficiency, replacement costs, value of excess days not pregnant, and cost of treatment had the greatest effect on returns from treatment. Herds with high conception rates and low replacement costs were likely to realize the least benefit from GnRH treatment at insemination. On the basis of our findings, we concluded that GnRH treatment at insemination is a profitable procedure under most herd conditions. Optimal treatment regimens for specific herds may best be determined by using herd performance and management data for calculating returns. PMID- 3045065 TI - Damage to bacteria by antibiotics in vitro and its relevance to antimicrobial chemotherapy: a historical perspective. PMID- 3045066 TI - The chemoprophylaxis of meningococcal infection. PMID- 3045068 TI - Inhibition of adherence of Candida albicans by conventional and experimental antifungal drugs. AB - We tested the effects of antifungal drugs on adherence of Candida albicans in vitro. Significant reduction of adherence occurred after 2 h incubation with amphotericin B, nystatin, miconazole, econazole, ketoconazole, chlorohexidine and ICI 195,739. Significant inhibition of candida adherence by 5-fluorocytosine and amorolfin required 18 h incubation. Combinations of amphotericin B with 5 fluorocytosine, miconazole, ICI 195,739 and amorolfin resulted in synergistic inhibition of adherence. Adherence is an important pathogenic mechanism in candida infections and interference with this process may represent a major component of the mode of action of antifungal drugs. PMID- 3045067 TI - Ceftazidime combined with mecillinam: serum bactericidal titres compared with in vitro synergy against gram-negative bacilli. AB - In a study of the possible interaction between mecillinam and ceftazidime against Gram-negative bacilli, ten volunteers received on separate days: ceftazidime 20 mg/kg iv in 15 min, mecillinam 10 mg/kg iv in 15 min, or the combination. Blood samples were obtained before and 1 and 6 h after the end of the infusion. Ten strains each of Klebsiella pneumoniae, Serratia marcescens, Citrobacter freundii, Salmonella spp. and Yersinia spp. and nine strains each of Acinetobacter spp., and Pseudomonas aeruginosa were selected. Most of the strains were resistant to ampicillin and cefazolin. Serum levels of ceftazidime and mecillinam were measured by bioassay. Serum bacteriostatic (SBS) and bactericidal (SBA) titration was done in microtitre plates in cation supplemented Mueller-Hinton broth and 50% human serum. Chequerboard titration was also studied to assess in-vitro synergy between ceftazidime and mecillinam in Mueller-Hinton broth with or without 50% serum. The mean serum concentrations (SD) were for mecillinam: 6.1 (1.7) at 1 h, and less than 0.3 at 6 h and for ceftazidime: 36.3 (5.5) at 1 h and less than 5 at 6 h. Identical concentrations were measured for the combination. By chequerboard titration, no synergy occurred for Acinetobacter spp. and Ps. aeruginosa, whereas it was observed in 37/60 (FIC) and 33/60 (FBC) of the strains of other species in Mueller-Hinton; from the strains showing synergy, 28/37 (FIC) and 30/33 (FBC) showed also synergy in Mueller-Hinton with 50% human serum. In SBS and SBA, on the other hand, the combination of mecillinam with ceftazidime showed an additive effect against most Enterobacteriaceae tested, synergy being shown for only 10-35% of tests.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3045069 TI - Topically applied vidarabine sodium phosphate in genital herpes infection. AB - One hundred and thirty-four patients with recurrences of genital herpes were randomized in a double-blind trial to receive self-applied topical therapy with either vidarabine sodium phosphate in an aqueous gel or placebo gel. No significant differences were noted between the two groups in the duration of viral shedding, in the time to healing of the treated recurrence, in the interval between recurrences, or in the number of recurrences. PMID- 3045070 TI - Ciprofloxacin treatment of recurrent Salmonella typhimurium septicaemia in a splenectomized and immunosuppressed patient. PMID- 3045072 TI - Cultured circular smooth muscle from the rabbit colon. AB - Although cultured vascular smooth muscle cells have been extensively characterized and investigated, there are very few studies of cultured intestinal smooth muscle cells. The aim of this study was to culture colonic smooth muscle (CSM) cells from the rabbit colon. Freshly isolated CSM cells from the circular muscle layer of the distal colon were prepared by collagenase digestion. In primary culture, CSM cells attached to the culture vessels by 48 to 72 h, proliferated by 3 to 7 d, and reached confluency by 14 to 17 d with a "hill-and valley" pattern. Spontaneous contractions were not observed at any time at 21 degrees or 37 degrees C. Confluent primary cultures were greater than 95% CSM cells, as identified by intensely positive immunofluorescent staining to smooth muscle actin-specific CGA7 and muscle-specific HHF-35 monoclonal antibodies. Transmission electron microscopy of freshly isolated and proliferating CSM cells revealed ultrastructural features consistent with smooth muscle cells. We successfully cultured CSM cells of the rabbit from freshly isolated cells and validated these CSM cells by electron microscopy and immunocytochemical staining. These highly pure primary cultures may be used to investigate numerous aspects of CSM cell metabolism and physiology. PMID- 3045071 TI - Malignant tumor cell lines produce interleukin-1-like factor. AB - Sixty-four malignant cell lines were examined for interleukin-1 (IL-1) activities in their conditioned medium using thymocyte and fibroblast proliferation assays. Sixteen cell lines showed high IL-1 activity. Comparison of these activities with human IL-1 showed similarity between some biological properties. However there was no correlation between cell origin and IL-1 activity. These results suggest the possibility that most malignant cells may produce an IL-1-like factor. PMID- 3045073 TI - Effects of enzyme isolation on the basal lamina of developing avian limb bud epithelia. AB - The developing avian limb bud is a classic example of an epithelial-mesenchymal interaction. Numerous attempts at maintenance of the epithelia in culture have been predominantly unsuccessful. The fate of the isolated epithelial sheet of the limb bud [including the apical ectodermal ridge (AER)] in culture may depend at least in part on the integrity of its basal lamina following isolation. In this study the distal epithelium of the stage 23 limb bud was isolated utilizing trypsin and Dispase II in a variety of procedures. The integrity of the basal lamina of limb epithelium immediately upon isolation and after 2 h in culture was determined by immunofluorescent staining for laminin, and electron microscopy. In epithelial sheets isolated with Dispase II a direct relationship was observed between maintenance of the extracellular matrix at isolation and the preservation of the tissue structure and cytoarchitecture following 2 h in culture. In contrast, there was an accelerated deterioration during incubation of the tissue isolated with trypsin, independent of isolation conditions and integrity of basal lamina after isolation. Short-term maintenance of limb bud epithelial structure and cytoarchitecture after enzymatic isolation seems correlative to the maintenance of extracellular matrix at the epithelial basal surface. PMID- 3045075 TI - The autistic child and family functioning: a developmental-family systems perspective. AB - Autism is a severe, long-term developmental disorder that potentially has substantial influence on different aspects of the family system. Principles from family-systems theory are considered as they relate to the autistic child within the family. A selective and critical review is presented of research findings on the influence of the autistic child on the functioning and interactions of family members, including parents, siblings, and the family as a whole. Research findings are also reviewed on resources associated with successful family adaptation to the autistic child. Suggestions are offered for improved research to assess the relationship between certain child variables and measures of family functioning. PMID- 3045076 TI - Clinical effects of fenfluramine on children with autism: a review of the research. AB - A review of research studies published to date on the effects of fenfluramine on children with autism is presented. The current status of the fenfluramine research on children with autism is assessed. The review analyzed the methodological aspects of the research, the toxicity of fenfluramine, and the relationship between fenfluramine, neurotransmitter activity, cognitive ability, and subsequent behavioral change. The review of published data indicated that fenfluramine had positive effects on the reduction of hyperactivity and stereotypic behaviors in 33% of the subjects. The best responders were children with the highest baseline IQs. The conclusions address the need for appropriate subgrouping of autistic syndromes, which may lead to identification of responders to pharmacological treatments. The need for further study of the possible long term adverse side effects of flenfluramine is noted. Further experimental research on the effects of fenfluramine on children with autism is endorsed. PMID- 3045074 TI - Improved media for normal human muscle satellite cells: serum-free clonal growth and enhanced growth with low serum. AB - We have developed a serum-free medium for clonal growth of normal human muscle satellite cells (HMSC). It consists of an optimized nutrient medium, MCDB 120, plus a serum-free supplement, designated SF, that contains epidermal growth factor (EGF), insulin, dexamethasone, bovine serum albumin, and fetuin. Fibroblast growth factor was needed with dialyzed fetal bovine serum (dFBS) as the only other supplement, but in media containing SF, it was only slightly beneficial, and was omitted from the final medium without significant loss. Clonal growth of HMSC in MCDB 120 plus SF is as good as with 15% serum and 0.5% chicken embryo or bovine pituitary extract. However, growth is further improved by use of a doubly-supplemented (DS) medium containing both SF and 5% dFBS. Clonal growth of HMSC in the DS medium far exceeds that in previous media with any amount of serum, and monolayer growth is at least equal to that in conventional media with higher levels of serum. Cells grown in these media exhibit little differentiation, even when grown to high densities. However, they retain the capacity for extensive fusion and synthesis of increased creatine kinase when transferred to a serum-free differentiation-promoting medium, such as Dulbecco's modified Eagle's medium plus insulin. All experiments were done with clonal cultures of HMSC to insure that observed growth responses were always those of muscle cells. PMID- 3045077 TI - Cloning and expression in Escherichia coli of isopenicillin N synthetase genes from Streptomyces lipmanii and Aspergillus nidulans. AB - beta-Lactam antibiotics such as penicillins and cephalosporins are synthesized by a wide variety of microbes, including procaryotes and eucaryotes. Isopenicillin N synthetase catalyzes a key reaction in the biosynthetic pathway of penicillins and cephalosporins. The genes encoding this protein have previously been cloned from the filamentous fungi Cephalosporium acremonium and Penicillium chrysogenum and characterized. We have extended our analysis to the isopenicillin N synthetase genes from the fungus Aspergillus nidulans and the gram-positive procaryote Streptomyces lipmanii. The isopenicillin N synthetase genes from these organisms have been cloned and sequenced, and the proteins encoded by the open reading frames were expressed in Escherichia coli. Active isopenicillin N synthetase enzyme was recovered from extracts of E. coli cells prepared from cells containing each of the genes in expression vectors. The four isopenicillin N synthetase genes studied are closely related. Pairwise comparison of the DNA sequences showed between 62.5 and 75.7% identity; comparison of the predicted amino acid sequences showed between 53.9 and 80.6% identity. The close homology of the procaryotic and eucaryotic isopenicillin N synthetase genes suggests horizontal transfer of the genes during evolution. PMID- 3045078 TI - Ethanolamine utilization in Salmonella typhimurium. AB - Ethanolamine can serve as the sole source of carbon and nitrogen for Salmonella typhimurium if vitamin B12 is present to serve as a cofactor. The pathway for ethanolamine utilization has been investigated in order to understand its regulation and determine whether the pathway is important to the selective forces that have maintained the ability to synthesize B12 in S. typhimurium. We isolated mutants that are defective in ethanolamine utilization (eut mutants). These mutants defined a cluster of genes located between purC and cysA at 50 min on the Salmonella chromosome. A genetic map of the eut region was constructed. Included in the map are mutations which affect ethanolamine ammonia lyase, the first degradative enzyme, and mutations which affect the second enzyme in the pathway, acetaldehyde dehydrogenase. Transcriptional regulation of the eut genes was studied by using eut-lac operon fusions created by insertion of Mu d lac. Transcription is induced by the simultaneous presence of ethanolamine and B12 in the growth medium. The eut genes constitute a single unit of transcription. One class of mutations located at the promoter-distal end of the eut operon prevent induction of transcription. PMID- 3045079 TI - Characterization of the rec-1 gene of Haemophilus influenzae and behavior of the gene in Escherichia coli. AB - The rec-1 gene of Haemophilus influenzae was cloned into a shuttle vector that replicates in Escherichia coli as well as in H. influenzae. The plasmid, called pRec1, complemented the defects of a rec-1 mutant in repair of UV damage, transformation, and ability of prophage to be induced by UV radiation. Although UV resistance and recombination were caused by pRec1 in E. coli recA mutants, UV induction of lambda and UV mutagenesis were not. We suggest that the ability of the H. influenzae Rec-1 protein to cause cleavage of repressors but not the recombinase function differs from that of the E. coli RecA protein. PMID- 3045080 TI - Nickel affects expression of the nickel-containing hydrogenase of Alcaligenes latus. AB - The effects of nickel on the expression of hydrogenase in the hydrogen-oxidizing bacterium Alcaligenes latus were studied. In the absence of added nickel, both hydrogenase activity, measured as O2-dependent H2 uptake, and hydrogenase protein, measured in a Western immunoblot, were very low compared with the levels in cells induced for hydrogenase in the presence of nickel. Hydrogenase activity and protein levels were dependent on the added nickel concentration and were saturated at 30 nM added Ni2+. The amount of hydrogenase protein in a culture at a given nickel concentration was calculated from the H2 uptake activity of the culture at that Ni2+ concentration. Between 0 and 30 nM added Ni2+, the amount of hydrogenase protein (in nanomoles) was stoichiometric with the amount of added Ni2+. Thus, all of the added Ni2+ could be accounted for in hydrogenase. Between 0 and 50 nM added Ni2+, all the Ni present in the cultures was associated with the cells after 12 h; above 50 nM added Ni2+, some Ni remained in the medium. No other divalent metal cations tested were able to substitute for Ni2+ in the formation of active hydrogenase. We suggest two possible mechanisms for the regulation of hydrogenase activity and protein levels by nickel. PMID- 3045081 TI - Starvation-induced cross protection against heat or H2O2 challenge in Escherichia coli. AB - Glucose- or nitrogen-starved cultures of Escherichia coli exhibited enhanced resistance to heat (57 degrees C) or H2O2 (15 mM) challenge, compared with their exponentially growing counterparts. The degree of resistance increased with the time for which the cells were starved prior to the challenge, with 4 h of starvation providing the maximal protection. Protein synthesis during starvation was essential for these cross protections, since chloramphenicol addition at the onset of starvation prevented the development of thermal or oxidative resistance. Starved cultures also demonstrated stronger thermal and oxidative resistance than did growing cultures adapted to heat, H2O2, or ethanol prior to the heat or H2O2 challenge. Two-dimensional gel electrophoresis of 35S-pulse-labeled proteins showed that subsets of the 30 glucose starvation proteins were also synthesized during heat or H2O2 adaptation; three proteins were common to all three stresses. Most of the common proteins were among the previously identified Pex proteins (J.E. Schultz, G. I. Latter, and A. Matin, J. Bacteriol. 170:3903-3909, 1988), which are independent of cyclic AMP positive control for their induction during starvation. Induction of starvation proteins dependent on cyclic AMP was not important in these cross protections, since a delta cya strain of E. coli K-12 exhibited the same degree of resistance to heat or H2O2 as the wild-type parent did during both growth and starvation. PMID- 3045082 TI - Common ancestry of Escherichia coli pyruvate oxidase and the acetohydroxy acid synthases of the branched-chain amino acid biosynthetic pathway. AB - A number of enzymes require flavin for their catalytic activity, although the reaction catalyzed involves no redox reaction. The best studied of these enigmatic nonredox flavoproteins are the acetohydroxy acid synthases (AHAS), which catalyze early steps in the synthesis of branched-chain amino acids in bacteria, yeasts, and plants. Previously, work from our laboratory showed strong amino acid sequence homology between these enzymes and Escherichia coli pyruvate oxidase, a classical flavoprotein dehydrogenase that catalyzes the decarboxylation of pyruvate to acetate. We have now shown this homology (i) to also be present in the DNA sequences and (ii) to represent functional homology in that pyruvate oxidase has AHAS activity and a protein consisting of the amino terminal half of pyruvate oxidase and the carboxy-terminal half of E. coli AHAS I allows native E. coli AHAS I to function without added flavin. The hybrid protein contains tightly bound flavin, which is essential for the flavin substitution activity. These data, together with the sequence homologies and identical cofactors and substrates, led us to propose that the AHAS enzymes are descended from pyruvate oxidase (or a similar protein) and, thus, that the flavin requirement of the AHAS enzymes is a vestigial remnant, which may have been conserved to play a structural rather than a chemical function. PMID- 3045083 TI - Antigenic relatedness and N-terminal sequence homology define two classes of periplasmic flagellar proteins of Treponema pallidum subsp. pallidum and Treponema phagedenis. AB - The periplasmic flagella of many spirochetes contain multiple proteins. In this study, two-dimensional electrophoresis, Western blotting (immunoblotting), immunoperoxidase staining, and N-terminal amino acid sequence analysis were used to characterize the individual periplasmic flagellar proteins of Treponema pallidum subsp. pallidum (Nichols strain) and T. phagedenis Kazan 5. Purified T. pallidum periplasmic flagella contained six proteins (Mrs = 37,000, 34,500, 33,000, 30,000, 29,000, and 27,000), whereas T. phagedenis periplasmic flagella contained a major 39,000-Mr protein and a group of two major and two minor 33,000 to 34,000-Mr polypeptide species; 37,000- and 30,000-Mr proteins were also present in some T. phagedenis preparations. Immunoblotting with monospecific antisera and monoclonal antibodies and N-terminal sequence analysis indicated that the major periplasmic flagellar proteins were divided into two distinct classes, designated class A and class B. Class A proteins consisted of the 37 kilodalton (kDa) protein of T. pallidum and the 39-kDa polypeptide of T. phagedenis; class B included the T. pallidum 34.5-, 33-, and 30-kDa proteins and the four 33- and 34-kDa polypeptide species of T. phagedenis. The proteins within each class were immunologically cross-reactive and possessed similar N-terminal sequences (67 to 95% homology); no cross-reactivity or sequence homology was evident between the two classes. Anti-class A or anti-class B antibodies did not react with the 29- or 27-kDa polypeptides of T. pallidum or the 37- and 30-kDa T. phagedenis proteins, indicating that these proteins are antigenically unrelated to the class A and class B proteins. The lack of complete N-terminal sequence homology among the major periplasmic flagellar proteins of each organism indicates that they are most likely encoded by separate structural genes. Furthermore, the N-terminal sequences of T. phagedenis and T. pallidum periplasmic flagellar proteins are highly conserved, despite the genetic dissimilarity of these two species. PMID- 3045084 TI - Cloning and sequencing of the Escherichia coli chlEN operon involved in molybdopterin biosynthesis. AB - The nucleotide sequence of a HinPI-HpaII restriction nuclease fragment which complemented a delta chlE strain of Escherichia coli was determined. Two open reading frames were deduced to be the structural genes for ChlE and ChlN proteins, which have molecular weights of 44,067 and 26,719, respectively. Both proteins were required for complementing a chromosomal deletion of the chlE locus. The chlE and chlN genes were transcribed from a common promoter, chlEp, located upstream of chlE. Transcriptional and translational signal sequences were recognized in this region. PMID- 3045085 TI - Molecular cloning of the Escherichia coli miaA gene involved in the formation of delta 2-isopentenyl adenosine in tRNA. AB - Escherichia coli mia strains were shown to lack delta 2-isopentenylpyrophosphate transferase activity, the first step in the synthesis of the 2-methylthio derivative of 6-(delta 2-isopentenyl) adenosine (ms2i6A). A double mutant, rpsL (Smp) miaA, was streptomycin dependent. The wild-type miaA gene was cloned by selecting for lambda recombinant bacteriophage which eliminated the streptomycin dependent phenotype and was subsequently recloned into plasmid vectors. The cloned miaA gene restored the ms2i6A modification to tRNA. The miaA gene mapped to 95 min on the E. coli map, and we propose the order mutL-miaA-hflA-purA. PMID- 3045086 TI - Effect of a mutation preventing lipid modification on localization of the pCloDF13-encoded bacteriocin release protein and on release of cloacin DF13. AB - The pCloDF13-encoded bacteriocin release protein (BRP; Mr 2,871) is essential for the translocation of cloacin DF13 across the cell envelope of producing Escherichia coli cells. Overproduction of this BRP provokes lysis (quasilysis) of cells. Construction and analysis of a hybrid BRP-beta-lactamase protein (BRP-Bla) demonstrated that the BRP contains a lipid modified cysteine residue at its amino terminus and is mainly located in the outer membrane. The significance of lipid modification for the localization and functioning of the BRP was investigated. Site-directed mutagenesis was used to substitute the cysteine residue for a glycine residue in the lipobox of the BRP and the BRP-Bla protein. The mutated BRP was unable to bring about the release of cloacin DF13 and could not provide the lysis (quasilysis) of host cells. However, the mutated BRP strongly inhibited the colony-forming ability of the cells, indicating that induction of the mutated protein still affected cell viability. In contrast to the wild-type BRP-Bla protein, the mutated BRP-Bla protein was mainly located in the cytoplasmic membrane, indicating that the mutation prevented the proper localization of the protein. The results indicated that lipid modification of the BRP is required for its localization and release of cloacin DF13, but not for its lethality to host cells. PMID- 3045087 TI - Structures of the promoter and operator of the glpD gene encoding aerobic sn glycerol-3-phosphate dehydrogenase of Escherichia coli K-12. AB - The nucleotide sequence of a 690-base-pair DNA segment containing the control region for the glpD gene encoding aerobic sn-glycerol-3-phosphate dehydrogenase was determined. An ATG translation initiation codon with an adjacent ribosome binding site was found which preceded an open reading frame continuing 61 codons to the end of the DNA that was sequenced. The start site for transcription, identified by using primer extension analysis, was located 42 base pairs upstream from the proposed Met start codon. The transcription start site was preceded by a region containing typical -10 and -35 sequences found in bacterial promoters. A binding site for the cyclic AMP-cyclic AMP receptor protein complex (identified by comparison with the consensus-binding sequence and verified by using DNase I footprinting) was located just upstream from the -35 sequence, centered at position -63. The interaction site for the glp repressor was identified by using DNase I footprinting. It consisted of a 49-base-pair region which started at the 10 sequence and continued to position +38. This region contained two directly repeated sequences, each possessing hyphenated dyad symmetry, which suggests that the operator is tandemly repeated. The presence of two adjacent operators may explain why expression of the glpD gene is the most sensitive to repressor when compared with expression of the other operons that are members of the glp regulon. PMID- 3045088 TI - Cloning and sequencing of a Shiga-like toxin type II variant from Escherichia coli strain responsible for edema disease of swine. AB - A Shiga-like toxin type II variant (SLT-IIv) is produced by strains of Escherichia coli responsible for edema disease of swine and is antigenically related to Shiga-like toxin type II (SLT-II) of enterohemorrhagic E. coli. However, SLT-IIv is only active against Vero cells, whereas SLT-II is active against both Vero and HeLa cells. The structural genes for SLT-IIv were cloned from E. coli S1191, and the nucleotide sequence was determined and compared with those of other members of the Shiga toxin family. The A subunit genes for SLT-IIv and SLT-II were highly homologous (94%), whereas the B subunit genes were less homologous (79%). The SLT-IIv genes were more distantly related (55 to 60% overall homology) to the genes for Shiga toxin of Shigella dysenteriae type 1 and the nearly identical Shiga-like toxin type I (SLT-I) of enterohemorrhagic E. coli. (These toxins are referred to together as Shiga toxin/SLT-I.) The A subunit of SLT-IIv, like those of other members of this toxin family, had regions of homology with the plant lectin ricin. SLT-IIv did not bind to galactose-alpha 1-4 galactose conjugated to bovine serum albumin, which is an analog of the eucaryotic cell receptor for Shiga toxin/SLT-I and SLT-II. These findings support the hypothesis that SLT-IIv binds to a different cellular receptor than do other members of the Shiga toxin family but has a similar mode of intracellular action. The organization of the SLT-IIv operon was similar to that of other members of the Shiga toxin family. Iron did not suppress SLT-IIv or SLT-II production, in contrast with its effect on Shiga toxin/SLT-I. Therefore, the regulation of synthesis of SLT-IIv and SLT-II differs from that of Shiga toxin/SLT-I. PMID- 3045089 TI - A hybrid toxin from bacteriophage f1 attachment protein and colicin E3 has altered cell receptor specificity. AB - A hybrid protein was constructed in vitro which consists of the first 372 amino acids of the attachment (gene III) protein of filamentous bacteriophage f1 fused, in frame, to the carboxy-terminal catalytic domain of colicin E3. The hybrid toxin killed cells that had the F-pilus receptor for phage f1 but not F- cells. The activity of the hybrid protein was not dependent upon the presence of the colicin E3 receptor, BtuB protein. The killing activity was colicin E3 specific, since F+ cells expressing the colicin E3 immunity gene were not killed. Entry of the hybrid toxin was also shown to depend on the products of tolA, tolQ, and tolR which are required both for phage f1 infection and for entry of E colicins. TolB protein, which is required for killing by colicin E3, but not for infection by phage f1, was also found to be necessary for the killing activity of the hybrid toxin. The gene III protein-colicin E3 hybrid was released from producing cells into the culture medium, although the colicin E3 lysis protein was not present in those cells. The secretion was shown to depend on the 18-amino-acid-long gene III protein signal sequence. Deletion of amino acids 3 to 18 of the gene III moiety of the hybrid protein resulted in active toxin, which remained inside producing cells unless it was mechanically released. PMID- 3045090 TI - Regulation of DNA superhelicity by rpoB mutations that suppress defective Rho mediated transcription termination in Escherichia coli. AB - The highly defective rho-15 mutant of Escherichia coli produces plasmid DNA that is 22% less negatively supercoiled than DNA from an isogenic wild-type strain (J. S. Fassler, G. F. Arnold, and I. Tessman, Mol. Gen. Genet. 204:424-429, 1986). We extended our measurements of plasmid superhelicity to additional rho mutants and to strains containing mutations that suppress rho transcription termination defects; the suppressor mutations were in the rpoB and the rho genes. The superhelicity of plasmid DNA was reduced by 11 and 10%, respectively, in the rho 702 and rho-201 mutants, both of which are less defective in Rho-mediated transcription termination than rho-15. Plasmid superhelicity was restored in all the suppressed rho mutants; in one rpoB mutant, plasmid DNA was even more negatively supercoiled than in rpoB+ cells, whether in a rho+ or rho mutant background. Suppression of rho mutants enabled them to maintain plasmids that could not be maintained in the mutants in the absence of the suppressor mutations. The results indicate that in addition to DNA gyrase, topoisomerase I, and Rho, RNA polymerase is also a determinant of DNA superhelicity, and its effect is modified by the Rho protein. We propose that Rho may increase the degree of DNA unwinding by the transcription complex, possibly at transcription termination sites. PMID- 3045091 TI - Transcriptional regulation of katE in Escherichia coli K-12. AB - Escherichia coli produces two distinct species of catalase, hydroperoxidases I and II, which differ in kinetic properties and regulation. To further examine catalase regulation, a lacZ fusion was placed into one of the genes that is involved in catalase synthesis. Transductional mapping revealed the fusion to be either allelic with or very close to katE, a locus which together with katF controls the synthesis of the aerobically inducible hydroperoxidase (hydroperoxidase II). katE was expressed under anaerobic conditions at levels that were approximately one-fourth of those found in aerobically grown cells and was found to be induced to higher levels in early-stationary-phase cells relative to levels of exponentially growing cells under both anaerobic and aerobic conditions. katE was fully expressed in air and was not further induced when the growth medium was sparged with 100% oxygen. Expression of katE was unaffected by the addition of hydrogen peroxide or by the presence of additional lesions in oxyR or sodA, indicating that it is not part of the oxyR regulon. When katF::Tn10 was introduced into a katE::lacZ strain, beta-galactosidase synthesis was largely eliminated and was no longer inducible, suggesting that katF is a positive regulator of katE expression. PMID- 3045094 TI - Cloning and nucleotide sequence of the oriT region of the IncI1 plasmid R64. AB - The nucleotide sequence at the oriT region of the IncI1 plasmid R64 was determined. A recombinant plasmid carrying a 141-base-pair R64 sequence was mobilized with a normal frequency, while a plasmid carrying only 44 base pairs of this R64 sequence was mobilized with a frequency 1/10 that of the original plasmid. The oriT region of the R64 plasmid contains two inverted-repeat sequences. PMID- 3045093 TI - Increased cell surface hydrophobicity of a Serratia marcescens NS 38 mutant lacking wetting activity. AB - The cell surface hydrophobicity of Serratia marcescens appears to be an important factor in its adhesion to and colonization of various interfaces. The cell surface components responsible for mediating the hydrophobicity of S. marcescens have not been completely elucidated, but may include prodigiosin and other factors. In the present report we have investigated the potential role of serratamolide, an amphipathic aminolipid present on the surfaces of certain S. marcescens strains, in modulating cell surface hydrophobicity. The hydrophobic properties of a serratamolide-producing strain (NS 38) were compared with those of a serratamolide-deficient mutant (NS 38-9) by monitoring the kinetics of adhesion to hexadecane. Serratamolide production was monitored by thin-layer chromatography and the wetting activity of washed-cell suspensions on polystyrene. Wild-type NS 38 cells were far less hydrophobic than the serratamolide-deficient mutant cells were; the removal coefficients were 48 min-1 for the mutant, as compared with only 18 min-1 for the wild type. The data suggest that the presence of serratamolide on S. marcescens cells results in a reduction in hydrophobicity, presumably by blocking hydrophobic sites on the cell surface. PMID- 3045092 TI - Sensitivity of some marine bacteria, a moderate halophile, and Escherichia coli to uncouplers at alkaline pH. AB - The inhibitory effects of uncouplers on amino acid transport into three marine bacteria, Vibrio alginolyticus 118, Vibrio parahaemolyticus 113, and Alteromonas haloplanktis 214, into a moderate halophile, Vibrio costicola NRC 37001, and into Escherichia coli K-12 were found to vary depending upon the uncoupler tested, its concentration, and the pH. Higher concentrations of all of the uncouplers were required to inhibit transport at pH 8.5 than at pH 7.0. The protonophore carbonyl cyanide m-chlorophenylhydrazone showed the greatest reduction in inhibitory capacity as the pH was increased, carbonyl cyanide p trifluoromethoxyphenylhydrazone showed less reduction, and 3,3',4',5 tetrachlorosalicylanilide was almost as effective as an inhibitor of amino acid transport at pH 8.5 as at pH 7.0 for all of the organisms except A. haloplanktis 214. Differences between the protonophores in their relative activities at pHs 7.0 and 8.5 were attributed to differences in their pK values. 3,3',4',5 Tetrachlorosalicylanilide, carbonyl cyanide m-chlorophenylhydrazone, 2-heptyl-4 hydroxyquinoline-N-oxide, and NaCN all inhibited Na+ extrusion from Na+-loaded cells of V. alginolyticus 118 at pH 8.5. The results support the conclusion that Na+ extrusion from this organism at pH 8.5 occurs as a result of Na+/H+ antiport activity. Data are presented indicating the presence in V. alginolyticus 118 of an NADH oxidase which is stimulated by Na+ at pH 8.5. PMID- 3045096 TI - In vitro insertion of leader peptidase into Escherichia coli membrane vesicles. AB - Leader peptidase is an integral protein of the Escherichia coli cytoplasmic membrane whose topology is known. We have taken advantage of this knowledge and available mutants of this enzyme to develop a genetic test for a cell-free protein translocation reaction. We report that leader peptidase inserted into inverted plasma membrane vesicles in its correct transmembrane orientation. We have examined the in vitro membrane assembly characteristics of a variety of leader peptidase mutants and found that domains required for insertion in vivo are also necessary for insertion in vitro. These data demonstrate the physiological validity of the in vitro insertion reaction and strengthen the use of this in vitro protein translocation reaction for the dissection of this complex sorting pathway. PMID- 3045095 TI - A gene encoding an SOS inhibitor is present in different conjugative plasmids. AB - In 9 of 20 conjugative plasmids of different incompatibility groups, including F and R100 (or R6-5), coexist two sequences which are homologous, respectively, to the gene psiB, which encodes an inhibitor of SOS induction, and to the gene ssb, which encodes a single-stranded-DNA-binding protein. PMID- 3045097 TI - Purification of Rts1 RepA protein and binding of the protein to mini-Rts1 DNA. AB - RepA protein, essential for the replication of plasmid Rts1, was purified, and its binding to mini-Rts1 subregions was examined by a DNase I protection assay. RepA protected the incI and incII iterons, a region immediately upstream of the repA promoter, and a 10-base-pair region located between the most external incII iteron and a GATC box. The protection was less efficient when preheated RepA was used. PMID- 3045099 TI - Differential effects of alprazolam and imipramine in generalized anxiety disorder: somatic versus psychic symptoms. AB - Some researchers have recently suggested that antidepressants may be superior to benzodiazepines in the alleviation of generalized anxiety. In a 6-week, double blind, parallel-design study with flexible dosage scheduling, the authors compared the effects of alprazolam with those of imipramine in 60 patients who had generalized anxiety disorder. On rating scales that contained both psychic and somatic symptoms, patients in both treatment groups improved to a similar degree after 2 weeks. However, alprazolam was more effective in attenuating somatic symptoms, and imipramine was more effective in attenuating psychic symptoms such as dysphoria and negative anticipatory thinking. The authors' results suggest that, in generalized anxiety, somatic symptoms and hyperarousal selectively respond to drugs acting on the gamma-aminobutyric acid system, whereas psychic symptoms respond to treatments affecting the noradrenergic or serotonergic systems. PMID- 3045098 TI - Nucleotide sequence of katG, encoding catalase HPI of Escherichia coli. AB - The gene katG, encoding catalase HPI of Escherichia coli, was sequenced, predicting a 726-amino-acid protein. The sequence was confirmed by identification of potential regulatory elements and amino acid sequencing of peptides. HPI shows no homology to other catalases. The distances between katG, metF, and ppc were defined. PMID- 3045100 TI - Preinfusion anxiety and laboratory-induced panic attacks in panic disorder patients. AB - Seventy panic disorder patients participated in a double-blind, placebo controlled, randomized infusion study in which sodium lactate and isoproterenol were used to induce panic anxiety. Patients who panicked during lactate, isoproterenol, and placebo infusions generally had higher preinfusion anxiety scores. These findings held true irrespective of the order the infusions were given. Stepwise multiple regression analyses comparing panickers with nonpanickers as the criterion variable revealed that the items "afraid of going crazy," "feeling unsteady," and "feeling paralyzed" on the Panic Description Scale had the highest predicting values. PMID- 3045102 TI - Hypocalcemia, hypoparathyroidism, and organic anxiety syndrome. AB - The author describes a case of functional hypoparathyroidism with hypocalcemia that developed in a patient after ablation of a parathyroid adenoma. The patient's clinical presentation was characterized by a severe anxiety state. The literature on the neuropsychiatric manifestations of hypoparathyroidism indicates that anxiety states are not uncommon in this condition and suggests that hypoparathyroidism should be considered in the differential diagnosis of organic anxiety syndrome. PMID- 3045101 TI - Further evidence for an association between affective disorders and anxiety disorders: review and case reports. AB - The relationship between anxiety and affective disorders continues to be debated in the literature. The authors report a case of identical twins concordant for attention deficit disorder and somnambulism but discordant for major depression and agoraphobia with panic attacks. The authors discuss the implications of this "finding in nature." Results from neuroendocrinological, family, and twin studies are also reviewed that either lend support or question the existence of a relationship between anxiety disorders and affective disorders. PMID- 3045103 TI - Baclofen-induced psychosis: two cases and a review. AB - The authors describe two patients who were being treated with baclofen for tardive dyskinesia. One patient developed withdrawal symptoms after baclofen was stopped, and the other developed a manic attach while receiving the drug. The authors review the literature concerning these side effects and explore the possible mechanisms responsible for these effects. PMID- 3045104 TI - Fungal infection associated with intravenous drug abuse: a case of localized cerebral phycomycosis. AB - The authors present a case of confusion and mood disturbance caused by a focal cerebral fungal (phycomycosis) infection in an otherwise healthy intravenous drug addict. A review of the literature found only 9 cases of phycomycosis with localized cerebral involvement. This report describes the sixth occurrence of phycomycosis in an intravenous drug addict (the fifth to localize in the basal ganglia). In addition to the human immunodeficiency virus, unusual infectious causes of confusion and mood disturbance may be increasing as the intravenous drug-using population expands. Recognition of the clinical features of a fungal infection in a high-risk population may lead to earlier diagnosis and more effective treatment of this uniformly fatal disease. The clinician should consider localized cerebral phycomycosis as a cause of confusion and mood disturbance in intravenous drug addicts, especially when there is evidence of basal ganglia involvement. PMID- 3045105 TI - Oral dyskinesia associated with buspirone use in an elderly woman. AB - Pronounced oral dyskinesia developed in an elderly woman with advanced dementia who had been treated with buspirone for 3 days. No abnormal motor movements had existed before buspirone use. At the time of the report 4 months after symptom onset, the dyskinesia persisted. The author discusses possible mechanisms of action. PMID- 3045106 TI - Pharmacological effects of serotonin reuptake inhibitors. AB - The effects of a series of tricyclic and non-tricyclic antidepressants on the transport of [3H]-serotonin into synaptosomal fractions from rat brain in vitro and in vivo are summarized. Differences in potency in vitro and in vivo would appear to be due partly to the presence of active metabolites, formed in vivo, that show different selectivities for the serotonin transport site. Evidence suggests that the initial reduction in serotonin turnover after acute administration of serotonin reuptake inhibitors is followed by a return to control values after the drug has been administered for at least 2 weeks. These changes in amine turnover are associated with normalization of both the serotonergic and beta-adrenergic receptor systems. Evidence suggests that changes in neurotransmitter receptor numbers and function of blood cells (platelets and lymphocytes) in depressed patients are possible state markers of the illness and that antidepressant efficacy may not be directly associated with specificity of amine reuptake inhibition. Studies also show that the bilaterally bulbectomized rat exhibits deficits in platelet and synaptosomal serotonin transport that resemble those shown in depressed patients. This animal model of depression may be useful not only for detecting putative antidepressants but also for studying the mode of action of such drugs during chronic administration. PMID- 3045107 TI - Clinical effects of serotonin reuptake inhibitors in the treatment of depressive illness. AB - The serotonergic hypothesis of depression has stimulated the development of a range of chemically diverse compounds that have an exclusive effect on this neurotransmitter for potential use as antidepressant drugs. The group of serotonin reuptake inhibitors are at various stages of clinical development. The authors review the efficacy, side effect profiles, and toxicity of zimelidine, fluvoxamine, and citalopram in detail and ifoxetine, fluoxetine, indalpine, paroxetine, and sertraline in brief. Evidence suggests that, compared with tricyclic antidepressants, these drugs may cause fewer anticholinergic effects and lower cardiotoxicity in the treatment of major depressive disorders. These advantages need to be weighed against an emerging pattern of gastrointestinal complaints, insomnia, and akathisia. The place of serotonin reuptake inhibitors in therapy can be fully evaluated only after further studies, particularly those involving long-term use. The adverse experience associated with zimelidine suggests that caution and vigilance should be exercised in future studies of agents in this class. These drugs will undoubtedly provide important insights into depressive illness and some anxiety disorders and may fulfill their initial promise. PMID- 3045108 TI - The potential role of serotonin reuptake inhibitors in the treatment of obsessive compulsive disorder. AB - The authors examine the role of the serotonergic system in the modulation or genesis of obsessive compulsive disorder and the potential for the use of the new serotonin (5-HT) reuptake inhibitors in its treatment. The data available are still preliminary, but are accumulating rapidly, and appear to hold promise in helping to understand this disorder and treat it more effectively. An overview of the data available is presented to assist in the planning of future research in this subject area. At least six major questions need to be addressed by future work: (1) What are the major hypotheses regarding the involvement of the serotonin system in the modulation or genesis of OCD? (2) Is there a serotonin system defect associated with OCD? (3) Is a serotonin system defect the underlying cause of OCD? (4) Are serotonin reuptake inhibitors effective in the treatment of OCD? (5) Are the effects of the serotonin reuptake inhibitors that are effective in treating OCD mediated via the serotonin system or via another mechanism? and (6) Are these drugs worth pursuing as treatment of OCD? PMID- 3045109 TI - Panic disorder: the potential role of serotonin reuptake inhibitors. AB - The authors review the role of the serotonergic system in panic disorder and the potential for use of the new serotonin (5-HT) reuptake inhibitors in treating panic disorder. Data currently available are only preliminary but suggest a future potential for this class of drug in the treatment of panic/anxiety disorders. The authors present a succinct and comprehensive overview of the data available, which aims to assist in the planning of future research in this area. PMID- 3045110 TI - Carbohydrate craving, mood changes, and obesity. AB - Carbohydrate craving can cause weight gain in affected people and is present in women with premenstrual syndrome (PMS) and persons with seasonal affective disorder (SAD). The neurotransmitter serotonin regulates carbohydrate intake; its precursor, tryptophan, enhances serotonin release. Animal studies have shown that serotonergic drugs decrease carbohydrate consumption. Three studies of the eating patterns of over 150 obese subjects have shown that carbohydrate craving occurred at specific times, that is, at 4 p.m. and 9 p.m. A serotonergic drug (D fenfluramine) has been shown to decrease carbohydrate consumption by 40%. Further dietary and pharmacological studies of PMS and SAD are needed to determine serotonin's involvement with symptoms of depressed mood, increased fatigue, and carbohydrate craving. PMID- 3045111 TI - Serotonin and its place in the pathogenesis of depression. AB - The authors review the literature in an attempt to evaluate the relationship between serotonin and depression. Animal studies show that the administration of tryptophan (the precursor of serotonin) increased serotonin synthesis and influenced behavior. Low plasma tryptophan levels have been found in patients with endogeneous depression. Postmortem studies have shown an association between lowered hindbrain serotonin levels and suicide among depressed persons. The decreased serotonin levels in blood platelets during depression mirrored the neuronal changes. Tricyclic antidepressants inhibited platelet serotonin uptake and reduced imipramine binding sites on the platelets. A positive correlation between depression rating scores and platelet aggregatory response has been reported. Serotonin stimulated release of prolactin and growth hormone, although the prolactin response was less marked in depression. A marker for depressive illness is still sought, but it is likely to be related in some way to serotonin. PMID- 3045112 TI - Pharmacology of sertraline: a review. AB - Sertraline is a member of a new class of psychotherapeutic agents that selectively inhibit serotonin reuptake in the brain. Animal studies have demonstrated that inhibition of serotonin reuptake leads to enhanced serotonergic neurotransmission and indirectly results in a down-regulation of beta adrenoceptors. The preclinical pharmacology of sertraline predicts antidepressant activity without accompanying anticholinergic, cardiotonic, or sedative side effects. Recent laboratory and clinical observations pertaining to body weight and obsessive compulsive disorder suggest the possibility of broader clinical indications for selective serotonin reuptake blockers such as sertraline. PMID- 3045113 TI - Acute effects of sertraline, amitriptyline, and placebo on the psychomotor performance of healthy subjects over 50 years of age. AB - A double-blind, placebo-controlled, crossover study in 12 subjects (greater than or equal to 50 years) compared the effects of single oral doses of sertraline (100 mg) and amitriptyline (50 mg) with placebo as assessed by psychomotor function testing. Unlike sertraline and placebo, amitriptyline increased tracking error severity and impaired digit/symbol substitution. Sertraline slightly improved flicker frequency recognition. Both active drugs caused subjective drowsiness, although amitriptyline's effect was greater and of longer duration. Both drugs impaired subjectively assessed performance. Sertraline caused nausea, and amitriptyline, dry mouth; sertraline tended to increase supine systolic blood pressure. The authors conclude that sertraline has a considerably less detrimental effect on psychomotor performance and may have a slight activating effect not found with amitriptyline. PMID- 3045114 TI - Reassessment of the translocation hypothesis by kinetic studies on hexose transport in isolated rat adipocytes. AB - The effect of insulin and factors which have insulin-like activity on the kinetic parameters of 3-O-methyl-D-glucose (MeGlc) transport in rat adipocytes were assessed. Carrier-mediated uptake of MeGlc was estimated by the difference in the amounts of [14C]MeGlc and L-[3H]glucose taken up in cells under equilibrium exchange conditions at 37 degrees C. The Km and Vmax values in basal cells were 17.4 mM and 0.24 nmol/10(6) cells/s, respectively. Removal of endogenous adenosine by adenosine deaminase resulted in a 26% decrease in the basal rate due to a slight increase in the Km (19.6 mM) and a decrease in the Vmax value (0.20 nmol/10(6) cells/s). The maximum concentration (10 nM) of insulin decreased the Km to approximately one-half of the basal (7.1 mM) concomitant with an 8.5-fold increase in the Vmax value (2.04 nmol/10(6) cells/s). Submaximal concentrations (50 and 150 pM) of insulin, N6-phenylisopropyladenosine (1 microM), mechanical agitation of cells by centrifugal force (160 x g), low temperature (15 degrees C), 12-O-tetradecanoylphorbol-13-acetate (1 microM), and hydrogen peroxide (10 mM) all decreased the basal Km value to a range of 13.5-7.3 mM, concomitant with a 1.7-7.4-fold increase in the Vmax. A possible explanation for the alterations in the kinetic parameters may be that insulin and other factors cause the translocation of the mobile low-Km glucose transporters from an intracellular site to the cell surface, where the stationary high-Km transporters are located. Thus, when the Km and Vmax values of the hypothetical high-Km transporters were assumed to be 20 mM and 0.20 nmol/10(6) cells/s, respectively, and the Km of the low-Km transporters was assumed to be 7 mM, the theoretical Km decreased from 20 to 7.5 mM as the Vmax of the low-Km transporters increased from near 0 to 2.0 nmol/10(6) cells/s. The relation between empirical Km and Vmax values as affected by several agents and conditions followed closely the relation predicted by the above two-transporter model. PMID- 3045115 TI - Localization of phosphorylation sites with respect to the functional domains of the mouse L cell glucocorticoid receptor. AB - Digestion of the rat liver glucocorticoid receptor with chymotrypsin results in the generation of a 42-kDa fragment which contains the steroid-binding and DNA binding domains and the antigenic site for the BuGR anti-glucocorticoid receptor monoclonal antibody, while digestion with trypsin generates a 15-kDa receptor fragment containing only the DNA-binding function and the BuGR epitope (Eisen, L.P., Reichman, M.E., Thompson, E.B., Gametchu, B., Harrison, R. W., and Eisen, H.J. (1985) J. Biol. Chem. 260, 11805-11810). In this paper, glucocorticoid receptor of mouse L cells that were grown in the presence of [32P]orthophosphate was digested with trypsin or chymotrypsin (either before or after immune purification with BuGR antibody) and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis, autoradiography, and Western blotting. The receptor is endogenously phosphorylated only on serine residues. Chymotrypsin digestion results in a 32P-labeled 42-kDa receptor fragment which contains steroid-binding, DNA-binding, and BuGR-reactive sites. Trypsin digestion generates a 27-kDa steroid-bound fragment (meroreceptor) which is not labeled with 32P and a 32P-labeled 15-kDa fragment which contains both the DNA-binding domain and the BuGR epitope. We have calculated that there are 4 times as many phosphate residues in the intact receptor than in the 42-kDa chymotrypsin fragment. From examination of 32P-labeled receptor fragments, we have deduced that one phosphate is located between amino acids 398 and 447, a region containing the BuGR epitope and about one-third of the DNA-binding domain, and the remaining three phosphates appear to be clustered just to the amino-terminal side of the BuGR epitope in a region defined by amino acids 313 to 369. Treatment of intact 32P-labeled receptor in cytosol with alkaline phosphatase removes these three phosphates, but it does not remove the phosphate from the DNA-binding-BuGR reactive fragment and it does not affect the ability of the transformed receptor to bind to DNA-cellulose. PMID- 3045116 TI - Two forms of RPO41-dependent RNA polymerase. Regulation of the RNA polymerase by glucose repression may control yeast mitochondrial gene expression. AB - We have identified two chromatographically separable forms of mitochondrial RNA polymerase from Saccharomyces cerevisiae which utilize different DNA templates. One form is only active in a nonselective assay utilizing a poly[d(A-T)] template. The other form selectively initiates from a mitochondrial promoter consensus sequence. Both enzymes can be extracted from yeast mitochondria and all components are encoded by nuclear genes. The possibility that these two activities represent core and holoenzyme forms of the multicomponent mitochondrial RNA polymerase is supported by our observation that both enzymes are absent from a strain bearing a disrupted copy of the RPO41 gene (Greenleaf, A. L., Kelly, J. L., and Lehman, I. R. (1986) Proc. Natl. Acad. Sci. U. S. A. 83, 3391-3399). The two enzyme activities are differentially regulated by carbon source; the nonselective enzyme is repressed during growth on glucose relative to the selective enzyme. The 5-fold increase in RNA polymerase activity on a nonrepressing carbon source correlates with the increased level of transcript production from mitochondrial DNA. These results suggest that the mitochondrial RNA polymerase and, in consequence, mitochondrial transcription are regulated by carbon catabolite control. PMID- 3045118 TI - Site-directed mutagenesis of mouse dihydrofolate reductase. Mutants with increased resistance to methotrexate and trimethoprim. AB - Site-directed mutagenesis was used to generate mutants of recombinant mouse dihydrofolate reductase to test the role of some amino acids in the binding of two inhibitors, methotrexate and trimethoprim. Eleven mutations changing eight amino acids at positions all involved in hydrogen bonding or hydrophobic interactions with dihydrofolate or one of the two inhibitors were tested. Nine mutants were obtained by site-directed mutagenesis and two were spontaneous mutants previously obtained by in vivo selection (Grange, T., Kunst, F., Thillet, J., Ribadeau-Dumas, B., Mousseron, S., Hung, A., Jami, J., and Pictet, R. (1984) Nucleic Acids Res. 12, 3585-3601). The choice of the mutated positions was based on the knowledge of the active site of chicken dihydrofolate reductase established by x-ray crystallographic studies since the sequences of all known eucaryotic dihydrofolate reductases are greatly conserved. Enzymes were produced in great amounts and purified using a plasmid expressing the mouse cDNA into a dihydrofolate reductase-deficient Escherichia coli strain. The functional properties of recombinant mouse dihydrofolate reductase purified from bacterial extracts were identical to those of dihydrofolate reductase isolated from eucaryotic cells. The Km(NADPH) values for all the mutants except one (Leu-22--- Arg) were only slightly modified, suggesting that the mutations had only minor effects on the ternary conformation of the enzyme. In contrast, all Km(H2folate) values were increased, since the mutations were located in the dihydrofolate binding site. The catalytic activity was also modified for five mutants with, respectively, a 6-, 10-, 36-, and 60-fold decrease of Vmax for Phe-31----Arg, Ile 7----Ser, Trp 24----Arg and Leu-22----Arg mutants and a 2-fold increase for Val 115----Pro. All the mutations affected the binding of methotrexate and six, the binding of trimethoprim: Ile-7----Ser, Leu-22----Arg, Trp-24----Arg, Phe-31--- Arg, Gln-35----Pro and Phe-34----Leu. The relative variation of Ki for methotrexate and trimethoprim were not comparable from one mutant to the next, reflecting the different binding modes of the two inhibitors. The mutations which yielded the greatest increases in Ki are those which involved amino acids making hydrophobic contacts with the inhibitor. PMID- 3045117 TI - Cloning of authentic human epidermal growth factor as a bacterial secretory protein and its initial structure-function analysis by site-directed mutagenesis. AB - A synthetic chimeric gene, coding for the human epidermal growth factor fused to the signal peptide of Escherichia coli alkaline phosphatase, was cloned into E. coli under the transcriptional control of the trp-lac (tac) promoter. Following induction with isopropylthiogalactoside, the secretion of the correctly processed protein product into the bacterial periplasm was detected and quantitated by its specific binding to the epidermal growth factor receptor. The purified protein was identical to authentic human epidermal growth factor in size, amino acid composition, primary sequence, receptor binding, and stimulation of receptor protein-tyrosine kinase activity. Based on interspecies homologies, structural considerations, and reported studies with peptide fragments, structure-function analysis was initiated with alterations of targeted amino acid residues by oligonucleotide-directed mutagenesis. The receptor binding affinity of each mutant, relative to the wild type, was measured by both radioreceptor competition and receptor tyrosine kinase stimulation assays. In general, the values obtained by the two methods were in agreement for each species of epidermal growth factor and followed the order: wild type greater than Glu24----Gly greater than Asp27--- Gly much greater than Pro7----Thr greater than Tyr29----Gly greater than Leu47--- His. The relatively low values obtained with the last two mutants suggest that Tyr29 and Leu47 may be important for the biological activity of human epidermal growth factor. PMID- 3045119 TI - Expression and processing of recombinant human terminal transferase in the baculovirus system. AB - Overproduction of human terminal transferase protein has now been accomplished by cloning the coding sequence of human terminal transferase into a baculovirus, where the expression of terminal transferase is under the control of the polyhedrin protein promoter. Two constructs were made, one producing a protein containing the entire terminal transferase fused to 12 amino acids from the NH2 terminus of the polyhedrin protein, and the other producing 58-kDa human terminal transferase. The terminal transferase levels expressed in cells infected with either recombinant baculovirus are around 10,000 units/10(7) cells at 48 h postinfection, about 200-fold greater than levels expressed in thymus and cultured lymphoblastoid cells. The chimeric polyhedrin/human terminal transferase protein produced in the infected insect cells has a molecular weight of about 60,000 while the nonfused recombinant human terminal transferase is identical in molecular weight to that present in human lymphoblastoid cells. Both forms of recombinant terminal transferase show immunological and enzymatic activity. When infected cells are pulse-labeled with [35S] methionine at 42-45 h postinfection, about 10% of newly synthesized protein is terminal transferase. Both forms of terminal transferase are phosphorylated in recombinant virus-infected cells as demonstrated by pulse-labeling infected cells with 32P-inorganic phosphate and isolation of labeled terminal transferase peptides by immunoprecipitation. PMID- 3045120 TI - Characterization of the sites of proteolytic activation of Newcastle disease virus membrane glycoprotein precursors. AB - The F1- and F2-polypeptide components of the fusion proteins and the hemagglutinin/neuraminidase proteins of the avirulent Queensland (V4) and virulent Australia-Victoria (AuV) strains of Newcastle disease virus have been isolated and subjected to extensive primary structural analysis including amino terminal sequence analysis and fast atom bombardment-mass spectrometry mapping. Nucleotide sequence analysis was performed on the gene which encodes the V4 hemagglutinin/neuraminidase protein. Signal peptidase cleavage was found to have occurred at the Ser31-Leu32 peptide bond of the primary translation products of the fusion protein genes. Activation cleavage of the V4 fusion protein precursor generated a sequence of -Gly-Lys-Gln-Gly84 at the carboxyl terminus of the F2 polypeptide and an amino-terminal sequence of the F1-polypeptide commencing with 86Leu-Ile-Gly-. The V4 hemagglutinin/neuraminidase protein gene was found to encode a primary translation product 45 amino acids longer at the carboxyl terminus than obtainable from the corresponding gene of the AuV strain (McGinnes, L. W., and Morrison, T. G. (1986) Virus Res. 5, 343-356). However, post translational proteolytic processing, exclusive to the primary translation product of the V4 hemagglutinin/neuraminidase protein gene, was found to have removed the last 42 residues of this carboxyl-terminal appendage. PMID- 3045121 TI - Calculation of the three-dimensional structure of Saccharomyces cerevisiae cytochrome b inserted in a lipid matrix. AB - Cytochrome b is an integral membrane protein, which forms the core of the ubiquinol-cytochrome c oxidoreductase (cytochrome bc1) complex. A computer-aided three-dimensional modeling procedure was carried out in four steps. First, the candidate hydrophobic helices were searched for throughout the protein primary sequence by a computer procedure based upon the method of Eisenberg; second, a secondary helical structure was imposed to the transmembrane peptides; third, the helical segments at a lipid-water interface were oriented, and finally the possible interactions between helices with similar properties were investigated. This procedure enabled the identification of nine hydrophobic segments, of which eight are membrane-spanning helices while one has amphipathic properties. Three hydrophilic receptor-binding domains were also identified. Based upon their hydrophobicity profiles, the transmembrane helices could be associated in pairs inside the lipid bilayer. In our folding model proposed for cytochrome b, all mutation sites are not only located on the same side of the membrane but are also in close proximity in the three-dimensional structure. Inhibitor resistance mutational sites which were recently characterized (di Rago, J.-P., and Colson, A.-M. (1988) J. Biol. Chem. 263, 12564-12570) have been located on this model. Moreover, the receptor-binding domains and the mutation sites are close neighbors in the three-dimensional spatial representation. PMID- 3045122 TI - Tris inhibits both proteolytic and oligosaccharide processing occurring in the Golgi complex in primary cultured rat hepatocytes. AB - Tris caused the distention of the Golgi cisternae in primary cultured rat hepatocytes and perturbed the functions occurring there. Proteolytic cleavage of precursors of both albumin and complement C3 was inhibited, whereas that of prohaptoglobin was not affected by Tris. These effects on the proteolytic cleavages resemble those of acidotropic amines (Oda, K., and Ikehara, Y. (1985) Eur. J. Biochem. 152, 605-609; Oda, K., Koriyama, Y., Yamada, E., and Ikehara, Y. (1986) Biochem. J. 240, 739-745). However, the effects of Tris significantly differed from acidotropic amines on the basis of its effects on the processing of N-linked oligosaccharides of glycoproteins. Both alpha 1-protease inhibitor and haptoglobin secreted from the Tris-treated cells were found to contain almost equal amounts of endo-beta-N-acetylglucosaminidase H-sensitive and -resistant oligosaccharides, whereas the glycoproteins from both the control and methylamine treated cells were resistant to the enzyme. The endo-beta-N-acetylglucosaminidase sensitive oligosaccharides were analyzed to be Man8-5GlcNAc by high resolution gel permeation chromatography, suggesting that trimming of alpha-mannose residues from the precursor Man9GlcNAc2 is incomplete in the Tris-treated cells. On the other hand, Tris did not significantly inhibit incorporation of radioactive monosaccharides (N-acetylglucosamine, galactose, and fucose) into the glycoproteins. However, two-dimensional gel electrophoresis in combination with neuraminidase digestion demonstrated that sialylation was markedly inhibited by Tris. Taken together, our results reveal that Tris inhibits not only the sialic acid addition which takes place in the trans Golgi region, but also the trimming step of high mannose-type oligosaccharides, which is thought to occur before glycoproteins reach the trans Golgi region. PMID- 3045123 TI - Proteinase-sensitive regions in the heavy chain of low molecular weight kininogen map to the inter-domain junctions. AB - Low molecular weight kininogen from human plasma was subjected to limited proteolysis with trypsin, chymotrypsin, elastase, and bromelain, and the resulting fragments of 20,000 or 40,000 Da were isolated. Amino-terminal sequence analysis of the fragments disclosed for the various proteinases eight independent cleavage sites distinct from the typical kallikrein cleavage sites flanking the kinin region. All the identified cleavage sites cluster in two stretches of 11-12 residues of the kininogen heavy chain. These short segments represent the primary attack sites for proteinases ("proteinase-sensitive regions") in the heavy chain portion of human low molecular weight kininogen. The amino acid sequences of the two proteinase-sensitive regions are mutually homologous; they are further characterized by the presence of a single copy each of the consensus tetrapeptide Cys-X-Gly-Cys known to form a narrow disulfide loop (Kellermann, J., Thelen, C., Lottspeich, F., Henschen, A., Vogel, R., and Muller-Esterl, W. (1987) Biochem. J. 247, 15-21). The proteinase-sensitive regions are located at the junctions of the three cystatin-like domains constituting the kininogen heavy chain. Proteolytic cleavage at the sensitive regions dissects the kininogen heavy chain and releases single domains of 20,000 Da and combined domains of 40,000 Da which can function as cysteine proteinase inhibitors. The presence of kininogen heavy chain domains in plasma samples under pathologic conditions suggests that cleavage of the proteinase-sensitive regions might also occur in vivo. PMID- 3045125 TI - A novel cleavage product of human complement component C3 with structural and functional properties of cobra venom factor. AB - The generation of two cleavage products of human C3, termed C3o and C3p, by incubation with a C3-cleaving protease isolated from cobra venom (Naja naja siamensis) is described. The venom protease removes the C3p fragment (Mr approximately 33,000) from the C3dg region of the C3 alpha-chain. The major cleavage fragment C3o (Mr approximately 140,000) contains the unaltered beta chain of C3 and two alpha-chain-derived polypeptides of Mr approximately 29,000 and Mr approximately 38,000, respectively. Amino-terminal amino acids sequence analysis of C3p and the three chains of C3o allowed the identification of the exact location of the two alpha-chain-derived fragments of C3o and the three cleavage sites of the venom protease. The chain structure of C3o resembles those of C3c and cobra venom factor. In contrast to C3c but like cobra venom factor (and C3b), C3o was found to support the activation of the serine protease Factor B by cleavage in the presence of Factor D and Mg2+ into Bb and Ba, generating an enzymatically active complex that is able to cleave a fluorogenic peptide substrate for C3 convertases. Since the only stretch of amino acid residues of C3o not present in C3c is the carboxyl terminus of the Mr approximately 29,000 chain of C3o, it is suggested that this region is important for the interaction with Factor B and convertase formation. PMID- 3045124 TI - Insulin action rapidly decreases multifunctional protein kinase activity in rat adipose tissue. AB - ATP-citrate lyase in vivo contains three phosphorylation sites on two tryptic peptides (peptides A and B). These phosphorylation sites are under hormonal control. Multifunctional protein kinase (MFPK) from rat liver phosphorylates peptide B on serine and threonine residues whereas cAMP-dependent protein kinase phosphorylates peptide A on a serine residue (Ramakrishna, S., and Benjamin, W. B. (1985) J. Biol. Chem. 260, 12280-12286). We now report that rat adipose tissue MFPK also phosphorylates serine and threonine residues of peptide B of ATP citrate lyase. When the activity of MFPK was assayed using partially purified (by chromatography on phosphocellulose) cytosol fractions from insulin-treated adipose tissue, it was found that MFPK activity was decreased by over 55%. This decrease in MFPK activity occurs at physiological concentrations of insulin (EC50 = 1 x 10(-10) M). Its onset is rapid and almost maximal at 5 min after the addition of insulin. Even when new protein synthesis is inhibited by cycloheximide, extracts from insulin-treated fat pads have less MFPK activity compared to the control. The insulin effect is maintained after further chromatography on a gel filtration column suggesting that the decrease in MFPK activity is not due to a low molecular weight inhibitor. The insulin-induced decrease in MFPK activity is due to a decrease in Vmax whereas the affinity of this enzyme toward ATP-citrate lyase or ATP is unchanged. PMID- 3045126 TI - Transcriptional regulation of c-myc oncogene expression by estrogen in hormone responsive human breast cancer cells. AB - Enhanced c-myc oncogene expression associated with peptide mitogen-stimulated cell growth is primarily a result of a post-transcriptional event involving increased mRNA stability. We have recently shown that estradiol stimulates c-myc expression in estrogen receptor-positive human breast cancer cells. In this report, we show that in estrogen-responsive MCF-7 cells, estradiol stimulated the c-myc gene exclusively at the transcriptional level, increasing c-myc mRNA transcription more than 10-fold within 20 min, while having no effect on the c myc mRNA half-life of 18 min. In addition, pretreatment of the cells with cycloheximide did not prevent induction of the c-myc oncogene, indicating a primary effect of estrogen. Furthermore, the elevated level of c-myc mRNA in estrogen-independent MDA-MB-231 cells was due primarily to a more stable c-myc mRNA with a half-life of 49 min, in the absence of enhanced transcription. These results indicate that different mechanisms of regulation of c-myc oncogene expression exist in hormone-dependent and -independent human breast cancer cells. PMID- 3045127 TI - Molecular analysis of plasmid DNA repair within ultraviolet-irradiated Escherichia coli. I. T4 endonuclease V-initiated excision repair. AB - The process by which DNA-interactive proteins locate specific sequences or target sites on cellular DNA within Escherichia coli is a poorly understood phenomenon. In this study, we present the first direct in vivo analysis of the interaction of a DNA repair enzyme, T4 endonuclease V, and its substrate, pyrimidine dimer containing plasmid DNA, within UV-irradiated E. coli. A pyrimidine dimer represents a small target site within large domains of DNA. There are two possible paradigms by which endonuclease V could locate these small target sites: a processive mechanism in which the enzyme "scans" DNA for dimer sites or a distributive process in which dimers are located by random three-dimensional diffusion. In order to discriminate between these two possibilities in E. coli, an in vivo DNA repair assay was developed to study the kinetics of plasmid DNA repair and the dimer frequency (i.e. the number of dimer sites on a given plasmid molecule) in plasmid DNA as a function of time during repair. Our results demonstrate that the overall process of plasmid DNA repair initiated by T4 endonuclease V (expressed from a recombinant plasmid within repair-deficient E. coli) occurs by a processive mechanism. Furthermore, by reducing the temperature of the repair incubation, the endonuclease V-catalyzed incision step has been effectively decoupled from the subsequent steps including repair patch synthesis, ligation, and supercoiling. By this manipulation, it was determined that the overall processive mechanism is composed of two phases: a rapid processive endonuclease V-catalyzed incision reaction, followed by a slower processive mechanism, the ultimate product of which is the dimer-free supercoiled plasmid molecule. PMID- 3045128 TI - Molecular analysis of plasmid DNA repair within ultraviolet-irradiated Escherichia coli. II. UvrABC-initiated excision repair and photolyase-catalyzed dimer monomerization. AB - In this study, a novel approach to the analysis of DNA repair in Escherichia coli was employed which allowed the first direct determination of the mechanisms by which endogenous DNA repair enzymes encounter target sites in vivo. An in vivo plasmid DNA repair analysis was employed to discriminate between two possible mechanisms of target site location: a processive DNA scanning mechanism or a distributive random diffusion mechanism. The results demonstrate that photolyase acts by a distributive mechanism within E. coli. In contrast, UvrABC-initiated excision repair occurs by a limited processive DNA scanning mechanism. A majority of the dimer sites on a given plasmid molecule were repaired prior to the dissociation of the UvrABC complex. Furthermore, plasmid DNA repair catalyzed by the UvrABC complex occurs without a detectable accumulation of nicked plasmid intermediates despite the fact that the UvrABC complex generates dual incisions in the DNA at the site of a pyrimidine dimer. Therefore, the binding or assembly of the UvrABC complex on DNA at the site of a pyrimidine dimer represents the rate-limiting step in the overall process of UvrABC-initiated excision repair in vivo. PMID- 3045129 TI - Genotoxic activities of benzamidine and its N-hydroxylated metabolite benzamidoxime in Salmonella typhimurium and mammalian cells. AB - The genotoxic potentials of benzamidine and benzamidoxime were determined to study the toxicological relevance of the metabolic N-oxygenation (N hydroxylation) of benzamidines to benzamidoximes. Benzamidoxime induced DNA single-strand breaks (in rat hepatocytes) and DNA amplification in SV40 transformed hamster cells. In the experiments performed, benzamidine itself was only marginally positive in the hepatocyte/DNA single-strand break assay. Since these cells possess an intact metabolization apparatus, the biological activities may be attributed to toxic and genotoxic metabolites formed by biotransformation. In the Salmonella typhimurium mutagenicity test (TA 98 and TA 100) benzamidoxime alone exhibited a low mutagenicity in the TA 98 strain in the presence of rabbit liver S-9 fractions. These results permit recognition of the metabolic N hydroxylation of benzamidines to benzamidoximes as a process to toxication. Indirect evidence for the formation of a glucuronide of benzamidoxime has been obtained from in vitro experiments, but it could not be established that this process was a decisive factor in the genotoxicity of benzamidoxime. PMID- 3045130 TI - Immunotoxins against solid tumors. AB - Antibody-toxin conjugates, termed immunotoxins, are currently being evaluated as potential new anticancer agents. The monoclonal antibodies that recognize antigens on the surface of tumor cells should deliver the toxins or the catalytic subunits of toxins to cancer cells. The catalytically active parts of the immunotoxins have to reach the cell cytoplasm where they inhibit protein synthesis. Immunotoxins against various solid tumors, including breast carcinoma and ovarian carcinoma, have been developed. In vitro, the activity of immunotoxins is affected by the number of target antigens on the cell surface, the internalization of the immunotoxins, the kind of toxin, the class of the antibody, the kind of linkage, and by other factors. Several problems arise with in vivo administration of immunotoxins. The short serum half-life of immunotoxins, due to their rapid hepatic uptake, decreases the number of immunotoxin molecules that reach the solid tumor. This, together with low tumor penetration by immunotoxins, could lead to low anti-tumor activity. Heterogeneity of tumors, immunogenicity of immunotoxins, and cross-reactivity of immunotoxins with normal tissues are other factors that might limit the clinical use of immunotoxins. It should be possible, however, to overcome these problems using methods that are already available or have yet to be developed. PMID- 3045131 TI - Increased amount of a mitochondria-associated 21 x 10(3) dalton protein in human gastrointestinal tumors. AB - Differences in the structure and number of mitochondria in tumor cells were found. Using isoelectrofocusing two-dimensional polyacrylamide gel electrophoresis which allows detection of alterations in the protein pattern of tumor mitochondria, we studied both quantitative and qualitative changes in the mitochondrial protein pattern of human gastrointestinal tumors and corresponding normal matrix tissues. One low molecular protein spot was found to be quantitatively changed in the tumors. The approximate molecular weight was 21 x 10(3) daltons and the pI value 5.7. PMID- 3045132 TI - Fetal type 2 pneumocytes form alveolar-like structures and maintain long-term differentiation on extracellular matrix. AB - We investigated the effects of reconstituted basement membrane (a crude extract of the Engelbreth-Holm-Swarm tumor) on type 2 pneumocyte differentiation during long-term culture. Cells were derived from mature 29 d fetal rabbits. Morphology was studied by light and electron microscopy. On thin gel, the cells initially segregated into clumps; they were cuboidal with apical microvilli and contained lamellar bodies, but dedifferentiated by 8 d. On thick gel, epithelial cells associated into spherical clusters surrounding a central lumen. These alveolarlike structures persisted at least 22 d. The cells were cuboidal and had lamellar bodies and intercellular tight junctions; they exhibited polarity, with apical microvilli facing the lumen, basally located nuclei, and gel matrix abutting the basal surface. In contrast, cells cultured on plastic formed colonies, then a monolayer, but dedifferentiated 5-7 d after plating. [14C]Acetate was used to label newly synthesized phospholipids. The amount of disaturated phosphatidylcholine (DSPC), expressed as a percentage of total phosphatidylcholine (PC), was used as an indicator of surfactant lipid production; percentage DSPC synthesized by cells cultured on thick gel did not change significantly, from 55 +/- 3 at 3 d, to 63 +/- 2 at 22 d in culture. DSPC synthesized by cells cultured on plastic decreased from 57 +/- 1% at 3 d to 45 +/ 2% at 22 d (p less than 0.001), which is consistent with the morphologic evidence of dedifferentiation. Synthesis of total PC compared with total phospholipid did not vary with either time in culture or substrate. This study emphasizes the importance of a complex extracellular matrix for maintenance of type 2 pneumocyte differentiation. The system should prove useful for studying the interaction of these cells with basement membrane, including the role of events occurring at the cell surface in modulating expression of a differentiated phenotype. PMID- 3045133 TI - Altered molecular properties of tubulin in a multidrug-resistant variant of Chinese hamster cells selected for resistance to vinca alkaloids. AB - The basis for the markedly altered intracellular binding of [3H]vincristine in a multidrug-resistant variant (DC-3F/VCRd-5L) of Chinese hamster lung cells (DC-3F) was investigated. Binding of [3H]vincristine by protein in cytosol derived from each cell type exhibited a differing requirement for GTP in MgCl2 containing buffer of low-ionic strength. Binding of [3H]vincristine occurred to cytosolic protein derived from both variant and parental DC-3F cells, but after removal of GTP, binding only occurred to cytosolic protein from parental cells regardless of the presence of added GTP. Although binding by cytosolic protein from parental DC 3F cells did not require GTP, the addition of 0.1 mM GTP increased by two-fold the rate and extent of binding. When cytosol from variant and parental DC-3F cells was incubated with low concentrations of [3H]vincristine in high-ionic strength buffer and analyzed by molecular-sieve HPLC, most of the protein binding [3H]vincristine in parentally derived cytosol eluted as Mr 110,000-115,000 daltons, corresponding to that for dimeric tubulin. The same binding species was not detected in cytosol derived from variant cells. However, these same fractions derived with both parental and variant cytosols contained tubulin as shown by SDS PAGE and immunoblotting. A smaller peak of [3H]vincristine binding and an amount of tubulin equal to that found in later fractions were found in the void volume during the same HPLC elution runs with cytosol from both variant and parental DC 3F cells. Evidence was also obtained for differences between parental and variant DC-3F cells in beta-tubulin isoforms following isoelectric focusing and immunoblotting. Parental-cell cytosol contains a single isoform of beta-tubulin. However, in variant cell cytosol the same isoform and, in addition, three more basic isoforms were found. These alterations in [3H]vincristine binding and in isoform compositions of beta-tubulin in variant versus parental DC-3F cells may have importance in regard to vincristine resistance in DC-3F cells. PMID- 3045134 TI - Identification of insulin receptors on the insulin-independent variant 1246-3A cell line. AB - 1246-3A cell line is an insulin-independent variant isolated from the adipogenic cell line 1246 which can proliferate in the absence of insulin, has lost the ability to differentiate, and secretes an insulin-related factor called IRF similar to pancreatic insulin and different from IGFs. In contrast, the parent adipogenic cell line 1246 is dependent on the presence of insulin to proliferate and differentiate in defined medium. In the present paper, we examined if the loss of response to insulin observed for 1246-3A cells was accompanied by alterations in the insulin receptor properties. Insulin binding and tyrosine kinase activity of insulin receptors isolated from 1246-3A cells and from the parent cell line 1246 were measured; 125I-insulin binding to intact cells was 75% lower for the 1246-3A cells than for the 1246 cells. This was due to a decrease in receptor number without major change in receptor affinity. However, when the cells were solubilized in 1% Triton X-100 and the insulin receptor was partially purified by chromatography on wheat germ agglutinin-agarose, a similar pattern of binding was observed for both cell lines. Down regulation of insulin receptors by insulin occurred in a dose-dependent fashion, which was similar for both cell lines. Phosphorylation experiments were performed by incubation of the partially purified insulin receptor with insulin and [gamma-32P]ATP. They indicated that insulin stimulated phosphorylation of the 95-kDa molecular weight beta subunit of the receptor, in a similar fashion for both cell types. These data suggest that the insulin-independent cell line 1246-3A does not possess a specific defect in the insulin receptor which alters both its binding and autophosphorylation properties and that the loss of response to insulin can be attributed to the fact the 1246-3A cells secrete IRF which bind to cell surface receptors and stimulate their proliferation. PMID- 3045136 TI - Rapid confirmation of enzyme-multiplied immunoassay test (EMIT) for phencyclidine positive urine samples with capillary gas chromatography-nitrogen-phosphorus detection. PMID- 3045135 TI - Regulation of neuroblast proliferation by hormones and growth factors in chemically defined medium. AB - Pure neuronal cultures prepared from 6-day-old embryonic chick brains incorporated [3H]-thymidine in serum-free medium up to the 4th day in culture. The addition of insulin any time within this culture period caused an increase in thymidine incorporation. This increase in [3H]-thymidine was correlated with an increase in cell number and percentage of labeling index. Triiodothyronine and endothelial cell growth factor were also active in stimulating [3H]-thymidine incorporation into chick neuroblasts. The effect of these trophic agents is unique since a variety of known mitogens tested were negative. PMID- 3045137 TI - Monitoring of cyclosporin in whole blood by reversed-phase liquid chromatography on a butyl column. PMID- 3045138 TI - Chromatographic analysis of the trinitrophenyl derivatives of insulin. AB - The insulin molecule was derivatised by reaction with trinitrobenzenesulphonic acid (TNBS), which is known to react predominantly with free primary amino groups. The products of the reaction were analysed by reversed-phase chromatography and by further derivatisation with dansyl chloride. Under the conditions of these experiments, TNBS was found to react preferentially with glycine at position A1. This finding is discussed in terms of the tertiary structure and immunogenicity of this derivative. PMID- 3045139 TI - Analysis of the amino acid and sugar composition of streptococcal cell walls by gas chromatography-mass spectrometry. AB - A procedure for determining the amino acid and sugar composition of streptococcal peptidoglycan-polysaccharide complexes by capillary gas chromatography-mass spectrometry (GC-MS) was established. Amino acids are analysed as butyl heptafluorobutyl derivatives and sugars as alditol acetates. These two different groups of compounds are derivatized independently but chromatography in both cases utilizes the same OV-1701 fused-silica capillary column which simplifies GC MS analysis. The butyl heptafluorobutyl procedure incorporates new pre- and post derivatization clean-up steps. Additionally, selected-ion monitoring MS allows amino acids to be readily analysed without interference from background noise. PMID- 3045140 TI - High-performance liquid chromatographic method for the determination of the esterase activity of subtilisin and kallikrein. PMID- 3045142 TI - Comparison in sensitivity of 10 HIV antibody detection tests by serial dilutions of Western blot-confirmed samples. AB - Nine ELISA tests and one particle agglutination test for the detection of HIV antibodies were analysed using serial dilutions of Western blot-confirmed positive serum samples. These samples were diluted in HIV antibody negative human serum in a range of 1:10 to 1:10,000 or 1:100,000. Remarkable differences in sensitivity were observed. Some tests were 10- to 100-fold more sensitive than others, and even differences of 1000-fold were detected. PMID- 3045141 TI - Detection of antibody to hepatitis B core antigen (anti-HBc) using a direct (antiglobulin) format and development of a confirmatory assay for anti-HBc. AB - A direct (antiglobulin) solid-phase enzyme immunoassay for the detection of antibody to hepatitis B core antigen (anti-HBc) is described. The assay utilizes recombinant hepatitis B core antigen as the solid-phase 'capture' reagent and a mixture of monoclonal antibodies specific for human IgG and IgM conjugated to horseradish peroxidase as the 'detector' reagent. The direct assay demonstrated excellent sensitivity and specificity when compared with a commercially available competitive enzyme immunoassay. The direct assay format lends itself to a confirmatory assay for anti-HBc by addition of monoclonal anti-HBc to the reaction mixture. Feasibility of the confirmatory assay for anti-HBc was demonstrated using specimens reactive for anti-HBc as documented by both the direct and competitive assays. PMID- 3045143 TI - Fasting decreases rates of noninsulin-mediated glucose uptake in man. AB - Although fasting decreases insulin-mediated glucose uptake (IMGU), its effect on noninsulin-mediated glucose uptake (NIMGU) is not known. To examine this issue we studied seven obese men [mean (+/- SD) age, 36 +/- 5 yr; weight, 91 +/- 13 kg] after an overnight fast (day 0) and 3 days (day 4) and 9 days (day 10) of total fasting and six normal weight men (age, 32 +/- 4 yr; weight, 73 +/- 6 kg) after an overnight and 3 days of fasting. To study NIMGU, somatostatin (0.16 micrograms/kg.min) was infused to create severe insulin deficiency and [3H]3 glucose to measure glucose disappearance (Rd), while serum glucose was sequentially clamped at a level of about 4.7 mmol/L for 180 min and about 11 mmol/L for an additional 100 min. The results from the last 60 min of each glycemic plateau were used for analysis. Under these conditions insulin action is absent and Rd = NIMGU. Since under conditions of euglycemic insulinopenia, NIMGU into noncentral nervous system tissues is negligible, and central nervous system (CNS) glucose uptake saturates at physiological glucose concentrations, it follows that at a glucose level of about 4.7 mmol/L, NIMGU reflects CNS glucose uptake and at about 11 mmol/L, NIMGU reflect CNS plus non-CNS tissues. Thus, non CNS NIMGU = NIMGU at 11 mmol/L - NIMGU at about 4.7 mmol/L. The obese subjects' mean weight fell to 88 +/- 5 kg on day 4 and 85 +/- 5 kg on day 10 (P less than 0.001 between all values). The mean basal serum glucose level fell from 5.3 +/- 0.1 on day 0 to 4.2 +/- 0.2 and 3.8 +/- 0.2 mmol/L on days 4 and 10, respectively (P less than 0.01 between all values). During insulinopenia plasma FFA and serum beta-hydroxybutyrate levels on day 10 were 3- and 30-fold higher than the basal prefast levels, respectively. Noninsulin-mediated glucose clearance at about 4.7 mmol/L did not change during fasting [0.0016 +/- 0.0001 (day 0) vs. 0.0016 +/- 0.0001 (day 4) and 0.0014 +/- 0.0001 L/kg.min (day 10); P = NS between all values]; at about 11 mmol/L noninsulin-mediated glucose clearance fell from 0.0016 +/- 0.0001 on day 0 to 0.0001 +/- 0.0001 dL/kg.min on day 10 (P less than 0.001). Results in the lean group were similar to those in the obese group after a 3-day fast.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3045145 TI - Insulin absorption and subcutaneous blood flow in normal subjects during insulin induced hypoglycemia. AB - We studied the effects of insulin-induced hypoglycemia on the absorption of 10 U 125I-labeled soluble human insulin injected sc in the thigh in 10 normal subjects. The disappearance of 125I from the injection site was followed by external gamma-counting. Subcutaneous blood flow (ATBF) was measured concomitantly with the 133Xe washout technique. The plasma glucose nadir [mean, 2.0 +/- 0.1 (+/- SE) mmol/L] occurred at 33 +/- 3 min and resulted in maximal arterial plasma epinephrine concentrations of approximately 6 nmol/L. From 30 min before to 60 min after the glucose nadir the [125I]insulin absorption rate was depressed compared to that during normoglycemia. The first order disappearance rate constants were reduced by approximately 50% (P less than 0.01) during the first 30-min interval after the glucose nadir. During the same period ATBF increased by 100% (P less than 0.05). The results suggest that in normal subjects the absorption of soluble insulin from a sc depot is depressed in connection with hypoglycemia, despite considerably elevated ATBF. PMID- 3045144 TI - Porcine and human insulin absorption from subcutaneous tissues in normal and insulin-dependent diabetic subjects: a deconvolution-based approach. AB - The mechanisms of sc insulin absorption are not understood, and models for interpreting in vivo data cannot be developed without gross simplification. To overcome this difficulty we developed a new approach which makes use of deconvolution analysis and does not require any model of the sc tissue. In five normal subjects and seven insulin-dependent diabetic (IDDM) patients endogenous insulin secretion was suppressed by means of a hypoglycemic glucose clamp procedure (approximately 2.8 mmol/L) sustained by a continuous insulin infusion (approximately 4 pmol/min.kg). A bolus injection of insulin (5.4 nmol) was administered iv, and plasma insulin concentrations were measured frequently for 2 h to assess iv insulin kinetics. Insulin then was injected sc in the abdominal region, and plasma insulin concentrations were measured for 8 h. Each subject was studied twice, with porcine and semisynthetic human insulin (Actrapid, Novo). The rate of insulin absorption was reconstructed by deconvolution from the plasma concentrations and iv insulin kinetic data. Linearity of the iv insulin kinetics, essential for deconvolution analysis, was confirmed by a dose-response study in the range of the measured concentrations (150-1800 pmol/L). In most instances, a two-compartment model was adequate to describe the iv response. The mean plasma insulin clearance rates were 15.5 +/- 1.9 (+/- SD) mL/min.kg (porcine) and 17.2 +/- 6.0 (human) in normal subjects and 20.7 +/- 8.8 (porcine) and 20.9 +/- 9.1 (human) in the IDDM patients. The rate of appearance of human insulin from sc tissue was faster than that of porcine insulin in both normal and IDDM subjects, but no significant differences were found in bioavailability, which was 55 +/- 12% (+/- SD; porcine) and 61 +/- 34% (human) in the normal subjects, and 84 +/- 28% (porcine) and 86 +/- 23% (human) in the IDDM patients. The rate of absorption and bioavailability were higher in the IDDM patients than in the normal subjects, a difference possibly related to increased sc blood flow in the IDDM patients. No differences were found with regard to glucose requirement values, normalized to plasma insulin concentrations, in agreement with the finding that the bioavailability of the two insulin species was similar. PMID- 3045146 TI - Immunopathologic studies of cutaneous lupus erythematosus. AB - The studies, as outlined above, strongly suggest that there may be several pathophysiologic mechanisms resulting in the development of cutaneous lupus lesions. It appears that all lupus lesions are associated predominantly with a T cell infiltrate. Based upon the studies of the neonatal lupus infants, it has been hypothesized that the U1RNP and Ro(SS-A) autoantibodies of maternal origin play a direct pathologic role in the genesis of the annular polycyclic SCLE lesions and this may be mediated by antibody-dependent cellular cytotoxicity mechanisms in which the antibody binds to the respective antigen present on the keratinocyte plasma membrane and the effector cells are T cells derived from the infants. Other studies, using direct immunofluorescence techniques, have demonstrated an association of cutaneous lupus lesions occurring in the presence of immunoglobulin and complement at the dermal/epidermal junction (positive lupus band test) in which the neoantigen of the complement membrane attack complex (C5b C9) is detected. These data have been interpreted as indicating that immunoglobulin and complement, perhaps in the form of immune complexes, may play a role in the pathogenesis of some cutaneous lupus lesions. Additional studies have determined that there is a substantial number of lupus patients with cutaneous disease, without antinuclear antibodies, who fail to demonstrate the deposition of immunoglobulin and complement at the dermal/epidermal junction. Furthermore, other studies have indicated that ultraviolet light is capable of inducing lesions in lupus patients that histologically are identical to those of cutaneous lupus erythematosus but that failed to demonstrate the deposition of the immunoglobulin and complement components. Since discoid lupus lesions demonstrate a preponderance of T cells, it has been proposed that some of these lesions are the direct result of a T-cell cytotoxic event. However, the nature of the autoantigens responsible for this putative T cell-mediated cytotoxic response is unknown at the present time. The role of ultraviolet light in the genesis of the cutaneous lupus lesions appears to involve, within the epidermis, the generation of autoantigen macromolecules which then react with autoantibodies or specific T cells of the lupus host. PMID- 3045147 TI - Susceptibility testing of anaerobic bacteria. PMID- 3045148 TI - Rapid diagnosis of influenza A and B by 24-h fluorescent focus assays. AB - Murine monoclonal antibodies directed against type-specific antigens of influenza A and B viruses have been shown to be useful diagnostic reagents for the detection of influenza viruses by immunofluorescence testing of nasopharyngeal cells. We have developed fluorescent focus assays utilizing these antibodies in cell culture chamber slides and shell vials for the rapid diagnosis of influenza A and B. Chamber slide assays were compared with virus isolation in 160 specimens from 135 patients with symptoms of influenza. Virus isolation was compared with immunofluorescence testing in 38 of the 160 specimens. Compared with virus isolation, 24-h cell culture chamber slide assays had a sensitivity of 75% and a specificity of 96%. Immunofluorescence testing of nasopharyngeal cells was only 38% sensitive and 91% specific. Shell vial assays were compared with virus isolation for 89 specimens. At 16 to 18 h postinoculation, the shell vial assay was 84% sensitive and 100% specific. We conclude that both chamber slide and shell vial assays are rapid, sensitive, and specific techniques for the diagnosis of influenza. PMID- 3045149 TI - Synthesis of viral proteins in polymorphonuclear leukocytes infected with influenza A virus. AB - Various reports have indicated that infection of polymorphonuclear leukocytes (PMNL) with influenza virus causes depression of their metabolic and chemotactic responses, but the effect the PMNL has on the life cycle of influenza virus has not been well defined. The studies reported here were undertaken to determine whether influenza virus could replicate within PMNL. Virus-infected and uninfected PMNL were labeled with [35S]methionine and analyzed by gel electrophoresis and fluorography for detection of newly synthesized proteins. Both host- and virus-specific proteins were produced within PMNL. By using indirect immunofluorescence techniques combined with flow cytometry, the expression of newly synthesized viral antigens was detected in virus-infected PMNL. Plaque assays on supernatant fluid from infected PMNL showed that infectious progeny were not produced, indicating that influenza virus infection of PMNL is abortive. PMID- 3045150 TI - Kinetics of delta antigen and delta antibody in acute delta hepatitis: evaluation with different enzyme immunoassays. AB - The kinetics of delta antigen (HDAg) and anti-delta antibody (anti-HD) was analyzed in 22 acute delta hepatitis infections (11 coinfections and 11 superinfections), with an enzyme immunoassay developed by Organon Teknika and with two commercially available assays: Deltassay, a test for HDAg from Noctech and Abbott anti-delta enzyme immunoassay, a test for anti-HD from Abbott Laboratories. In seven cases, HDAg was detected with the Organon assay but not with Deltassay. These discrepancies were not related to the type of hepatitis delta virus infection. All samples from these patients taken beyond week 4 of illness were found anti-HD positive with both the Organon and Abbott anti-HD assays. These data reflect the lack of sensitivity of Deltassay. In no instance were HDAg and anti-HD present simultaneously when tested with the Organon assays. On the contrary, 10 sera among the 28 that were HDAg positive with the Organon assay also were found anti-HD positive with the Abbott test. We suspected that the test procedure recommended by Abbott (a one-step competitive assay) may have yielded false-positive anti-HD results when HDAg present in the sera reacted with peroxidase-labeled anti-HD of the kit. To determine the specificity of the simultaneous presence of HDAg and anti-HD, these 10 sera were retested with the Abbott anti-HD assay, but by a modified two-step procedure that avoided contact between sera and labeled antibody. For six sera a negative result was obtained with the second procedure, suggesting that a false anti-HD reaction occurred with the standard test procedure. For four sera a positive result with both procedures was indicative of the simultaneous presence of HDAg and anti-HD. In conclusion, assay for HDAg was found very convenient for the early diagnosis of acute hepatitis delta virus infections. Seroconversion to anti-HD could be used for a late diagnosis 2 to 5 weeks after the beginning of illness. However, anti-HD results obtained with one-step competitive assays have to be interpreted carefully in HDAg-positive sera. PMID- 3045151 TI - Congo red binding and salt aggregation as indicators of virulence in Shigella species. AB - Smooth strains of Shigella dysenteriae type 1, Shigella flexneri, Shigella boydii, and Shigella sonnei which form pigmented colonies (Pcr+) on Congo red agar were virulent in the Sereny test. Smooth variants unable to bind Congo red (Pcr-) were avirulent. Measurements of dye uptake from solution showed that S. dysenteriae type 1 bound the most dye, followed in order of uptake by S. flexneri, S. boydii, and S. sonnei. Using the salt aggregation test (SAT) to determine cell surface hydrophobicity, we found the same order of species. The SAT could not, however, detect differences in surface properties between Pcr+ and Pcr- pairs of isogenic smooth strains. Enteroinvasive Escherichia coli strains used in the study showed SAT and Congo red-binding properties which were similar to those of the S. flexneri strains. A direct correlation was found between pigment-binding ability and the presence of the large 140-megadalton plasmid in S. flexneri, enteroinvasive E. coli, and S. boydii but not in S. dysenteriae type 1 or S. sonnei strains. Congo red interacted with outer membranes and outer membrane proteins of S. dysenteriae type 1 but not with lipopolysaccharides. However, rough mutants of Shigella species deficient in lipopolysaccharides bound Congo red and formed pigmented colonies, showing that dye binding as a virulence assay may be misinterpreted in such cases. There was complete correlation of the Pcr+ phenotype with virulence in the smooth strains in this study, suggesting that Congo red binding can be utilized as a quick and reliable alternative to the Sereny test. PMID- 3045152 TI - Clinical and microbiologic features of Shigella and enteroinvasive Escherichia coli infections detected by DNA hybridization. AB - To determine the clinical and microbiologic features of Shigella and enteroinvasive Escherichia coli (EIEC) infections, we investigated 410 children with diarrhea and 410 control children without diarrhea who were seen at Children's Hospital, Bangkok, Thailand, from January to June 1985. Shigella spp. were isolated from 96 (23%) and EIEC were isolated from 17 (4%) of 410 children with diarrhea and from 12 (3%) and 6 (1%) of 410 control children, respectively. The isolation rates of both pathogens increased with age and peaked in children 3 to 5 years old from whom Shigella spp. were isolated from 38% and EIEC were isolated from 9%. Shigella spp. were isolated from 52% and EIEC were isolated from 7% of 91 children with bloody diarrhea and from 15 and 3% of 319 children with nonbloody diarrhea. Fifteen (65%) of 23 EIEC were lactose positive, and all belonged to recognized EIEC serotypes. Among children with diarrhea, the stool blots of 76% of 17 children infected with EIEC, 45% of 96 children infected with Shigella spp., and 1% of 297 culture-negative children hybridized with the 17 kilobase EcoRI digestion fragment of pRM17, a recombinant plasmid containing DNA derived from the 140-megadalton Shigella flexneri plasmid. Although EIEC colonies can be reliably detected by DNA hybridization, detection by stool blot is less sensitive. Shigella spp. and EIEC are important causes of endemic diarrhea among children greater than 1 year old in Thailand. PMID- 3045154 TI - Comparison of three rapid methods for detection of antibodies to streptolysin O and DNase B. AB - Three commercial systems were compared for ability to detect antibodies to streptolysin O (ASO) and DNase B (ADB). Streptozyme (Wampole Laboratories, Cranbury, N.J.) exhibited high sensitivity (100%) for detecting ASO but low sensitivity for ADB (22.2%). The LeapStrep (Organon Teknika, Malvern, Pa.) and Check-Spectra (Diagnostic Technology, Hauppauge, N.Y.) tests had low sensitivities for detecting ASO (35.3 and 21.4%, respectively) and ADB (22.2 and 33.3%, respectively). PMID- 3045156 TI - SIGNAL blood culture system in clinical practice. PMID- 3045153 TI - Characterization of flagella purified from enterohemorrhagic, vero-cytotoxin producing Escherichia coli serotype O157:H7. AB - Escherichia coli of the serotype O157:H7 has recently been isolated in human fecal specimens in association with sporadic cases and outbreaks of hemorrhagic colitis and with the hemolytic uremic syndrome. The aim of this study was to characterize the flagellin protein subunit constituents of flagellar filaments from E. coli O157:H7 strain CL-56. Flagellin isolated from a reference Salmonella enteritidis strain was used for comparison. Flagella were dissociated by incubation of bacteria under acidic conditions, centrifugation, and differential ammonium sulfate precipitation. Reconstituted flagellar filaments were demonstrated by three complementary methods: transmission electron microscopy, antigenic reactivity with H7 antiserum by a dot blot immunoassay, and immunogold localization of antiserum raised to the purified antigen to intact flagella on whole E. coli O157:H7. On sodium dodecyl sulfate-polyacrylamide gels flagellin proteins from E. coli O157:H7 demonstrated an apparent Mr of 66,000. The isoelectric point of E. coli O157:H7 flagellin was 5.42. By immunoblotting, H7 flagellin proteins were shown to be immunogenic. They induced a systemic immune response both in rabbits challenged with whole bacteria and in a human previously infected with E. coli O157:H7. PMID- 3045155 TI - Influence of culture medium pH and proteinase inhibitors on extracellular proteinase activity and cell growth of Sporothrix schenckii. AB - Sporothrix schenckii produces two extracellular proteinases in albumin- or collagen-supplemented unbuffered liquid medium. Proteinase I had an optimal pH of 6.0, and its activity was strongly inhibited by chymostatin. Proteinase II had an optimal pH of 3.5, and its activity was strongly inhibited by pepstatin. Speculating that these two proteinases are key enzymes for fungal growth, we investigated the influences of culture medium pH and either or both of the proteinase inhibitors pepstatin and chymostatin on the cell growth of S. schenckii. In buffered medium at a pH of 3.5, an optimal pH for proteinase II, only proteinase II activities were observed, while at pH 6.0, an optimal pH for proteinase I, only proteinase I activities were observed. However, there was no cell growth inhibition. These results suggested that the regulation of the production of the two proteinases is dependent on environmental pH. The addition of pepstatin or chymostatin to the culture medium did not inhibit the cell growth of S. schenckii, but the addition of both inhibitors at a concentration of 10 micrograms/ml strongly inhibited growth. These results suggested that at least one of the two proteinases was expressed to allow fungal growth in albumin supplemented media. The indirect fungistatic action of the proteinase inhibitors, which inhibit proteolysis, may be applied to a topical therapeutic agent in vivo. PMID- 3045158 TI - Detection of ampicillin resistant Haemophilus influenzae in United Kingdom laboratories. AB - Susceptibility of Haemophilus influenzae clinical isolates to ampicillin reported by 23 laboratories, using a variety of methods, was compared with results obtained following retesting at The London Hospital Medical College. Beta lactamase production was not detected on initial isolation in 25 of 157 isolates (16%) found to be positive on retest. One hundred beta lactamase negative isolates, which gave reduced zone diameters (less than 20 mm) around 2 micrograms discs and required 1-64 mg/l ampicillin for inhibition, were detected at The London Hospital. Eighty five of these had been reported as sensitive to ampicillin by the laboratories of origin. Many of these 100 isolates showed reduced susceptibility to other beta lactam antibiotics. Accurate detection of non-enzymic reduced susceptibility to ampicillin may emerge as an important guide to the likely sensitivity of H influenzae isolates to the enzyme stable beta lactams. PMID- 3045159 TI - Smooth muscle pseudotumours: a potentially confusing artefact of rectal biopsy. AB - An artefactual smooth muscle lesion was found in seven of 500 consecutive rectal biopsy specimens. The lesions had the deceptive appearance of a genuine tumour although none of the patients with the lesion had presented with a rectal mucosal swelling. The morphology of the lesion and its poor reproducibility under experimental conditions suggested that it was an artefact of the biopsy procedure: it was easily reproduced in resected specimens of large bowel using punch or basket forceps but not when using flat forceps. The presence of the lesion seems to depend on the type of forceps used rather than on differences in deployment and seems to be caused by avulsion of the superficial part of the muscularis propria and its incorporation into the tissues included in rectal biopsy specimens. PMID- 3045157 TI - Haematological abnormalities in human immunodeficiency virus (HIV) disease. AB - Peripheral blood and bone marrow changes are commonly seen in disease associated with human immunodeficiency virus (HIV). This annotation aims to summarise these changes and to suggest possible factors entailed in their occurrence. PMID- 3045160 TI - Laboratory investigation of paraproteinaemia. PMID- 3045161 TI - New rapid latex agglutination test for diagnosing Trichomonas vaginalis infection. AB - A newly developed latex agglutination test for Trichomonas vaginalis infection was compared for sensitivity, specificity, efficiency, and positive and negative predictive values with microscopy, culture, and an enzyme linked immunosorbent assay (ELISA) in the diagnosis of 395 women attending a genitourinary medicine clinic. T vaginalis infection was diagnosed in 42 (11%) women. The sensitivities of both the latex agglutination test and the ELISA were 95% compared with 74% for microscopy and 76% for culture. The latex test was specific and showed no cross reaction with a wide range of other genital tract infections. The latex agglutination test can detect antigen in both soluble and insoluble forms, and as it is simple to perform, can be undertaken during routine examination without recourse to special equipment or training. Further evaluation is required. PMID- 3045162 TI - Glycocalyx in virulent and avirulent strains of Shigella flexneri. PMID- 3045163 TI - Topical tretinoin in the treatment of aging skin. AB - We review the use of topical tretinoin (all-trans-retinoic acid) in the treatment of aging skin. We have found topical tretinoin capable of improving aged appearing skin in both a double-blind, vehicle-controlled trial and our clinic patients. The most impressive improvement occurs after application of tretinoin 0.1% cream for 8 to 12 months. Side effects have been limited to a mild, transient, and clinically insignificant burning sensation in the eyes and mild irritation of tretinoin-exposed skin. PMID- 3045164 TI - Liver toxicity of retinoid therapy. AB - Vitamin A metabolism involves storage in the liver. Hypervitaminosis A results in liver abnormalities, including fibrosis and cirrhosis. Ito cells are increased and promote fibrogenesis, which results in cirrhosis. Retinoids (Accutane and Tegison) are used clinically for the treatment of a variety of skin diseases. Since retinoids are analogs of vitamin A, their potential to produce liver disease is reviewed. Animal and human studies of liver function tests suggest some abnormalities in the liver in about 25% of patients treated. Liver biopsy studies have included isolated case reports and two retrospective and one prospective liver biopsy study of retinoids in humans. Although some increase in histologic liver changes have been noted, most liver biopsy specimens showed no change or improvement. Retinoids do not appear to produce consistent toxic liver abnormalities. PMID- 3045165 TI - Ocular effects of oral retinoids. AB - Knowledge of the ocular side effects of oral retinoids is important when treating patients with these drugs. The side effects include blepharoconjunctivitis, dry eye syndrome, cutaneous photosensitivity, contact lens intolerance, refractive changes, papilledema and pseudotumor cerebri, corneal opacities, and abnormal retinal function. The signs, symptoms, and management of these ophthalmologic effects are discussed. PMID- 3045166 TI - Papular elastorrhexis: a variant of connective tissue nevus. Case reports and review of the literature. AB - Two unrelated young women with papular elastorrhexis have recently been seen at the University of Iowa Hospitals and Clinics. This entity, recently described in a single patient, is a distinct variant of a connective tissue nevus. Although the histologic features of papular elastorrhexis may mimic those of other connective tissue nevi, clinical characteristics allow differentiation. The clinical and histologic characteristics of papular elastorrhexis, as well as its differential diagnosis, are discussed. PMID- 3045167 TI - Fertility of postpartum dairy cows after administration of gonadotropin-releasing hormone and prostaglandin F2 alpha: a field trial. AB - Our objective was to examine further the potential profertility effects of gonadotropin-releasing hormone and prostaglandin F2 alpha in postpartum Holstein cows. Reproductive performance was monitored in 843 cows milked thrice daily. One group of cows (n = 218) was untreated, while three groups received either 100 micrograms gonadotropin-releasing hormone administered once between d 11 and 25 (n = 211); 25 mg prostaglandin F2 alpha given once between d 11 and 25 (n = 215); or 25 mg prostaglandin F2 alpha given once between d 25 and 40 postpartum (n = 190). No profertility effects were detected in cows, regardless of their health status during the periparturient period, except cows given gonadotropin-releasing hormone between d 11 and 18 had shorter intervals to first estrus and to first service than controls. Cows with reproductive disorders (abnormal health status) in the concurrent lactation had longer intervals from calving to conception after receiving gonadotropin-releasing hormone between d 18 and 25 or prostaglandin F2 alpha between d 33 and 40. Abnormal health status adversely affected every reproductive trait measured. Early postpartum treatments with either hormone failed to improve reproductive performance of dairy cows, in contrast to several reports of profertility effects for gonadotropin-releasing hormone, and a few reports for prostaglandin F2 alpha. PMID- 3045168 TI - Penetration of gel and solution etchants in occlusal fissures. AB - The major clinical advantage of a phosphoric acid gel etchant is its superior control during placement. However, its viscosity can impede penetration into occlusal fissures, resulting in adequate etching and decreased pit-and-fissure sealant retention. This study examined the penetration of gel and liquid phosphoric acid etchants in occlusal fissures. The depth of fissure penetration of the acids was investigated using paired, sectioned samples and evaluated with a scanning electron microscope. The etch pattern was measured from the base of the fissure to the first demonstrable evidence of etched enamel. A variable pattern of etched enamel was present at the base of the fissure to no pattern observable at distances of 15 microns from the base. A frequent finding was that debris partially or totally blocked the fissure orifice, preventing acid penetration. Utilizing a paired t-test, no statistically significant difference (0.5 less than p less than 0.8) in the fissure penetration of gel or liquid phosphoric acid etchants could be shown. PMID- 3045169 TI - Ectodermal dysplasia with partial anodontia: prosthetic treatment with implant fixed prosthesis. AB - In cases with severe hypodontia, it is necessary to make a treatment plan in collaboration with different disciplines in dentistry, with the treatment plan formed in such a way that many possibilities for treatment could be used in the future. Partial and full dentures combine simple methods with good esthetic and functional results in the growing individual. They also afford time to assess the patient with regard to more extensive restorative treatment. Patients suffering from severe hypodontia have so far been very dependent on their remaining teeth. The case reported covers a twenty-year period of dental treatment, and shows a new possibility provided by osseointegrated implants. PMID- 3045170 TI - Radicular cysts of primary teeth mimicking premolar dentigerous cysts: report of three cases. AB - In three cases, the clinical diagnosis of dentigerous cyst was disproved by surgical exploration. In all other cases reviewed from a thirteen-year period, the clinical diagnosis of radicular cyst from an infected primary tooth was verified by surgery and histological examination. PMID- 3045171 TI - The problems associated with substituting composite resins for amalgam: a status report on posterior composites. PMID- 3045173 TI - The nature of the periapical lesion--a review of 1108 cases. PMID- 3045172 TI - Resin-bonded fixed partial dentures: a controlled clinical trial. PMID- 3045174 TI - A study of the clinical fit of cast cobalt-chromium clasps. PMID- 3045175 TI - 1987 Kreshover lecture. Gene regulation in the development of oral tissues. AB - The regulatory processes associated with tooth formation are being investigated by the identification of when, where, and how cell adhesion molecules (CAMs), substrate adhesion molecules (SAMs), dentin phosphoprotein, enamel gene products, and intermediate cementum products are expressed during sequential developmental stages of morphogenesis, cytodifferentiation, dentin, enamel and cementum extracellular matrix (ECM) formation, and biomineralization. Instructive and permissive signaling is required for both morphogenesis and cytodifferentiation based upon in vitro organotypic culture studies in serumless, chemically-defined medium. Intrinsic developmental instructions, independent of exogenous growth factors, mediate tooth morphogenesis from the initiation of the dental lamina through crown and initial root development. Recent progress using recombinant DNA methods has advanced descriptions of several dental structural genes. The complete nucleic acid sequence for mouse amelogenin has been defined. This sequence is located on the mouse X chromosome and on the human X and Y chromosomes. This discussion summarizes recent results using experimental embryology, recombinant DNA technology, and immunocytology in the context of instructive epithelial-mesenchymal interactions associated with epithelial differentiation into ameloblasts, ectomesenchyme differentiation into odontoblasts, and dentin and enamel ECM biomineralization. The tooth organ provides opportunities at several levels of biological organization to investigate cellular, molecular, and developmental processes. PMID- 3045176 TI - Parameters that affect in vitro bonding of glass-ionomer liners to dentin. AB - The purpose of this study was to evaluate the effects of two concentrations of poly (acrylic acid) (10 and 25%), three treatments (untreated, passive conditioning, and active conditioning), and two storage conditions (24 hours in 37 degrees C water and thermal cycling) on the in vitro tensile bond strength of three commercial glass-ionomer liners to human dentin. Bond strengths to untreated dentin after storage for 24 hours ranged from 19.0 to 21.7 kg/cm2 for Glasionomer Base Cement, Cement/Liner, and Ketac-Bond, but dropped to a range of 4.9 to 9.7 kg/cm2 after thermal cycling. Active conditioning with 10% acid resulted in bond strengths after 24-hour storage that ranged from 23.5 to 44.0 kg/cm2, compared with values from 21.7 to 38.0 kg/cm2 with active conditioning using 25% acid. Active conditioning with 10% acid resulted in bond strengths after thermal cycling that were in the range of 15.8 to 27.4 kg/cm2 and were 80 to 320 percent higher than values resulting from passive conditioning under these conditions. Active conditioning with 10% acid for 30 seconds produced a bond strength for Glasionomer Base Cement of 44.0 kg/cm2, compared with a bond strength of 28.7 kg/cm2 for a 10-second active conditioning. Qualitative analysis of scanning electron photomicrographs showed that dentin tubules were opened to a greater extent by active conditioning with 25% acid than by passive conditioning with 10% acid. PMID- 3045177 TI - Oxidation and ceramic coatings on 80Ni20Cr alloys. AB - Additives and/or impurities in NiCr alloys would be expected to affect the oxidation and adherence of dental glasses. Oxidation characteristics and bonding of several glasses to a commercial impure 80Ni20Cr alloy and laboratory-prepared purer alloys were thus studied at 1000 degrees C. The commercial alloy formed a porous and buckled single-layer oxide scale with lenticular voids at the interface. The scale was penetrated by glass which formed a chemical bond at the alloy/glass interface, resulting in excellent adherence with fracture in the glassy phase. The purer alloy formed a complex multilayer oxide scale to which the glass bonded but did not penetrate; the assembly was not satisfactory, since fracture occurred in the oxide scale. PMID- 3045178 TI - Composition of human plaque fluid. AB - The composition of pooled resting plaque fluid was determined in four groups of college-age students (18-22 years), each composed of 50 individuals, who abstained from oral hygiene for 36 hours and did not eat or drink for at least one hour prior to plaque collection. Plaque samples from each group were pooled under mineral oil in small centrifuge tubes and centrifuged at 37,000 g for one hour at 4 degrees C. Supernatants were then combined under mineral oil and centrifuged at 5000 g (4 degrees C) for 15 minutes. In general, the inorganic composition of plaque fluid in the four groups was quite similar and in agreement with values reported by other investigators, but quite different from those of saliva or serum. The mean composition was: Ca, 7.07 +/- 0.51 mmol/L; P, 23.2 +/- 5.3 mmol/L; Na, 18.6 +/- 2 mmol/L; K, 85.1 +/- 5.3 mmol/L; Mg, 3.9 mmol/L; Cl, 42.8 +/- 9 mmol/L; F, approximately 0.004 mmol/L; pH, 5.69 (5.63-6.01). Acetate, propionate, succinate, butyrate, lactate, and formate were determined in two samples analyzed, with acetate and propionate being the predominant acids found. It was also demonstrated, through the titration of one of the plaque fluid samples, that the observed buffer capacity in plaque fluid was mostly related to its organic acid composition. It was noted, however, that when the initial pH in plaque fluid exceeded 6.5, phosphate contributed significantly to the buffer capacity. The contribution of other soluble species (proteins, peptides, amino acids) to the observed buffering in plaque fluid appeared to be small.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3045179 TI - Fluctuating odontometric asymmetry in an urban South African black population. AB - The magnitude of fluctuating odontometric asymmetry is reported for a group of 106 urban South African Blacks by use of re-scaled asymmetry values and Euclidean map analyses. When these results were contrasted with those reported for South African Caucasoids and Paraguayan Lengua Indians, Blacks were found to be significantly more asymmetric. It is suggested that the disproportionately high levels of dental asymmetry may be ascribed to the high disease and malnutrition burden of South African Blacks and to decreased individual buffering ability. The present study failed to support Garn's model of x-chromosomal odontogenetic buffering in females, and also failed to confirm significant arcadal differences in the magnitude of fluctuating odontometric asymmetry. PMID- 3045180 TI - The crescentic ellipse revisited. PMID- 3045181 TI - Wound healing for the dermatologic surgeon. AB - An understanding of the basic science of cutaneous wound repair is essential to the dermatologic surgeon for the management of the postoperative wound. This review discusses the stages of wound healing and then applies these principles to the preoperative, intraoperative, and postoperative management of the surgical patient. PMID- 3045182 TI - The use of composite auricular grafts in nasal reconstruction. AB - Composite auricular grafts are extremely valuable in head and neck reconstruction. They are most commonly used in reconstruction of the ear, eyelid, and nose. Composite grafting provides a relatively simple technique of reconstruction with excellent cosmetic and functional results. The advantage of a one-stage reconstruction is particularly valuable to the patient. PMID- 3045183 TI - [Glycosaminoglycans: their structure and metabolism]. PMID- 3045184 TI - [Intracellular extralysosomal hydrolysis]. PMID- 3045185 TI - Absorption of ultrasound by mammalian ovaries. AB - The transient thermoelectric method was employed to determine the ultrasonic absorption coefficient of excised bovine, canine, feline, murine, ovine, and porcine ovaries at 1 MHz. The dynamics of the organ yields significant variations with physiological stage and structure. Values ranged from 0.017 cm-1 for the follicle to 0.050 cm-1 for the corpus luteum, largely reflecting macromolecular content. Little interspecies variation was observed. PMID- 3045186 TI - Alert glove use. PMID- 3045187 TI - The presidents. Frank P. Bowyer 1977-1978. PMID- 3045188 TI - Dentistry on stamps (Purkinje). PMID- 3045189 TI - A solitary cauliflower-like nodule on the mucosal surface of the lower lip. AB - The incidence of oral condylomata acuminata and simultaneous genital condylomata is not known. A review of the literature implies that the disease is rare, but it is more likely that it is only rarely reported. As the virus is autoinoculable and transmissible, the etiologic factor in this case of oral condylomata acuminatum is most likely orogenital contact. This case report serves as a reminder also that oroanogenital contact expands the anatomic range of sexually transmitted diseases. PMID- 3045190 TI - Tissue specificity of premature aging in diabetes mellitus. The role of cellular replicative capacity. PMID- 3045191 TI - Isoelectric focusing as the crow flies. AB - The evolution of isoelectric focusing is traced back over the years, from a somewhat shaky origin to present-day immobilized pH gradients. Four generations of methodology are classified and discussed: (A) Kolin's approach, consisting of a two-step technique, generation of a pH gradient by diffusion followed by a rapid electrokinetic protein separation; (B) Svensson-Rilbe's approach, consisting of creating a pH gradient in an electric field by utilizing as buffers a multitude of carrier ampholytes, i.e. of amphoteric species possessing good buffering capacity and conductivity at their pI; (C) immobilized pH gradients, by which non-amphoteric buffers and titrants (acrylamido weak acids and bases), titrated around their pK values, are grafted (insolubilized) onto a polyacrylamide gel matrix and (D) mixed-bed carrier ampholyte-Immobiline gel, by which a soluble, carrier ampholyte generated pH gradient coexists in the same matrix with an insoluble, Immobiline generated, pH gradient. PMID- 3045193 TI - Is the aminoterminal propeptide of type III procollagen degraded in the liver? A study of type III procollagen peptide in serum during liver transplantation in pigs. AB - The aminoterminal propeptide of type III collagen was monitored in serum during liver transplantation in nine pigs. The aim was to investigate whether removal of the liver causes any changes in the serum concentration of the propeptide. Another connective tissue component, hyaluronan, a glycosaminoglycan known to be degraded in the liver endothelial cells, was also measured. Removal of the liver caused a significant increase in the concentration of the intact propeptide as well as of hyaluronan. Gel filtration confirmed the increase in the amount of intact propeptide. However, another large propeptide-related antigen, eluted near the void volume, appeared in the antigen profile during the anhepatic phase. This peak probably represents the propeptide still attached to the collagen molecule (pN collagen). The findings indicate that the liver is involved in the degradation of the propeptide and of larger propeptide-holding proteins. PMID- 3045192 TI - Insulin degradation into monocytes from normal subjects: a high performance liquid chromatographic analysis. AB - In this work the fate of A14-125 I-insulin inside human cells has been investigated by the complementary use of gel permeation and reversed-phase high performance liquid chromatography to obtain a better resolution of the cell processed radioactive material resulting from the internalization of labeled insulin. Mononuclear leukocytes from 12 normals were incubated with pure A14-125 I insulin at 37 C and internalized radioactivity was characterized after 2, 15 and 60 min. Nearly 14% of intracellular radioactivity was associated to materials with a molecular weight of approximately 300,000. The remaining 86% had a molecular weight lower than 20,000. High molecular weight material showed an elution profile very similar to that obtained from purified human placental insulin receptor and was partially precipitable with antireceptor antibody. The reversed phase high performance liquid chromatography analysis of the low molecular weight material showed two main peaks corresponding to 125 I and A14 125 I-insulin and three intermediate peaks, a, b, c, accounting for about 8% of the recovered radioactivity. By increasing the incubation time of A14-125 I insulin with monocytes a decrease of insulin peak (2 min: 38 +/- 18%; 15 min: 25 +/- 11%; 60 min: 6 +/- 4%) and a corresponding increase of iodide peak was observed. Immunoprecipitability with anti-insulin antibody was 0% for iodide and a peaks, 60% for peak b, 78% for peak c and 90% for A14-insulin peak. Our results show that intracellular insulin degradation procedes rapidly and in a time dependent manner and that this process produces insulin derivatives which partially retain the immunological properties of intact A14-125 I insulin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3045194 TI - Diagnosis of acute rejection in liver transplantation. AB - Eleven acute rejections were found in 9 patients with liver transplantation due to end-stage liver cirrhosis. The rejections were diagnosed with fine-needle aspiration biopsy (FNAB) giving the cellular picture of immunoactivation in the liver graft when compared to a simultaneous sample of peripheral blood. s Alkaline phosphatase and s-bilirubin increased within 1 week after onset of rejection in 7 and 10 cases, respectively. s-Alanine amino-transferase and b ammonium were of no value in the diagnosis of acute rejection. A core biopsy was obtained only in a case of severe liver damage, mainly to estimate the need for retransplantation. One year after grafting, 6 out of 7 cirrhotic patients are well, all with normal liver function. Two have died of sepsis. One patient died from pulmonary metastases of occult liver carcinoma 6 months after the transplantation. FNAB seems helpful in detecting early acute rejection and also excluding such an event in the liver graft. PMID- 3045195 TI - Pathophysiology of ascites and functional renal failure in cirrhosis. PMID- 3045196 TI - Sex hormones and chronic liver disease. PMID- 3045197 TI - Medical museum notes. PMID- 3045198 TI - Neonatal sepsis. PMID- 3045199 TI - 19th century medicine in Grant County, land of the Miami Indians. PMID- 3045201 TI - Bibliography of the current world literature in hypertension. PMID- 3045203 TI - An epidemiological study of plasma renin activity in schoolchildren in Japan: distribution and its relation with family history of hypertension. AB - Plasma renin activity (PRA) was determined in a group of 610 Japanese schoolchildren aged 10-14 years in order to investigate the relationship between PRA distribution and a family history of hypertension. Plasma renin activity was higher in the subjects with a family history of hypertension (FH+) than in those without a family history of hypertension (FH-). Systolic blood pressure (SBP) was also higher in the FH+ group than in the FH- group. The FH- group showed a significant negative correlation between PRA and SBP (boys, r = -0.254, P less than 0.01; girls, r = -0.225, P less than 0.01), whereas the FH+ group showed no correlation between PRA and blood pressure. These results suggested that schoolchildren with a family history of hypertension might have an enhanced renin aldosterone (R-A) system, resulting in elevation of blood pressure. PMID- 3045200 TI - Lecithinase activity of Proteus vulgaris. AB - Lecithinase production is described as a new biochemical property of P. vulgaris strains grown in a selective agar medium containing brilliant green, crystal violet and lecithin (BCL agar), the authors' own modification of egg-yolk culture medium. By using this BCL agar as a medium inhibiting the swarming growth of P. vulgaris cultures the authors succeeded in identifying 12 lecithinase-positive strains among the P. vulgaris isolates obtained from patients with Crohn's disease. Of 50 P. mirabilis strains tested in parallel none gave the positive test for lecithinase production in this medium. PMID- 3045202 TI - Double-blind randomized, crossover trial of calcium supplementation in essential hypertension. AB - In a double-blind, randomized, placebo-controlled, crossover trial, 23 middle aged patients with mild to moderate essential hypertension were given an oral calcium supplement (1 g/day) for 8 weeks. At the end of this period, eight patients continued with this treatment for an additional 2 weeks but were also given 0.5 micrograms/day of 1,25-(OH)2 vitamin D3. In the 21 patients who completed the study, arterial pressure during the calcium-supplemented phase was almost identical to that of the placebo phase. In eight patients, mean arterial pressure (MAP) had changed by greater than 5 mmHg at the end of the calcium supplemented period, compared with the end of the placebo phase (six patients showed an increase in MAP and two a decrease). Changes in arterial pressure were unrelated to age, plasma ionized calcium, parathyroid hormone (PTH), plasma renin activity (PRA), plasma aldosterone, 24-h urinary calcium, sodium and potassium and were only weakly related to body weight. In the eight patients who continued with the treatment of calcium plus 1,25-(OH)2 vitamin D3 after the 8-week study period, arterial pressure changed very little and not significantly. These results do not support the suggestion that calcium supplements lower arterial pressure in middle-aged subjects with mild to moderate essential hypertension. PMID- 3045204 TI - [Role of Chlamydia trachomatis in male urethritis. Analysis of 2,000 cases of male urethritis]. AB - A study involving of 2,000 cases of urethritis revealed the presence of Chlamydia trachomatis in 44% of patients. Following a pathophysiological review, the strongly suggestive clinical picture of infection by this microorganism is emphasized. This study stresses two precise points by dealing with: firstly, the importance of the choice of technique for demonstration of the presence of the bacteria, and its reliability; secondly, the value of bacteriological evidence of the infection in order to treat not only the patient but also the partner(s) and to subsequently confirm the treatment as being effective. PMID- 3045206 TI - [Bilateral testicular Leydig cell tumor. Procedure to follow]. AB - The authors report one case of a bilateral testicular Leydig cell tumor in a man of 29 years old. There are few cases of such tumors reported in the literature. Gynecomastia force the patient to consult his doctor. His hormonal profile is practically normal; however, the serum estradiol is at the limit superior of normal range, and serum testosterone at the limit inferior. Testicular palpation is normal. It is the scrotal ultrasonography which confirms the diagnosis of testicular tumor. The scrotal ultrasonography has to be performed in every patient with unexplained gynecomastia. There is no metastasis. Before the treatment, the sperm conservation is performed (his sperm is normal). Our surgical sequence for this man without children was the next one: 1. inguinal orchiectomy was performed at the side of bigger tumor. Histological diagnosis was the benign Leydig cell tumor; 2. one month later, an inguinal orchidotomy at the other side was performed and the tumor palpable of 9 mm was removed. The extemporaneous biopsy confirmed the same diagnosis as at the other side; 3. one year later, there is no metastasis and the woman of our patient becomes pregnant. PMID- 3045205 TI - [Abscess of the kidney. Apropos of 21 cases]. AB - Renal abscess is rather an uncommon lesion caused predominantly by an urogenic infection. The authors report 21 cases in a period of 10 years. They describe and evaluate the actual investigation possibilities which are accurate enough to lead to the right diagnosis of renal abscess in a good rate. The evolution of the therapeutic modes is studied, with a special reference to the percutaneous drainage which is to substitute the classical surgery of this abscess. For the authors, treated in the right time and in the right way, renal abscess is nowadays a benign lesion, preserving a good prognosis for the kidney and the patient. PMID- 3045207 TI - Ectopic vasa deferentia in an infant with the prune belly syndrome. PMID- 3045208 TI - [Simple cysts of the testis in children. Apropos of 2 cases]. AB - Reporting two cases of simple testicular cysts in children, the authors analyse these rare benign tumors (9 other cases in literature). The cystic dilatation first affect the rete testis, it can be partial (simple cyst) or extensive (multicystic dysplasia). The primitive perturbation would affect the mesonephrotic formations, with a defect of junction between the two different embryologic formations of the testicle and its excretor system. These tumors are revealed by a large isolated testicle, histology only makes the diagnosis. Despite their benignity, simple orchidectomy is the save treatment; enucleation is justified only when the opposite testicle is also injured. PMID- 3045209 TI - Characteristics of dermal invasion in experimental cutaneous candidiasis of leucopenic mice. AB - The course of experimental cutaneous Candida albicans infections produced in mice made leucopenic by the administration of cyclophosphamide was compared to that in untreated animals. In the latter, neutrophils characteristically infiltrated the area of infection and the organisms were virtually always confined to the epidermis. However, even though many fewer foci of infection were associated with neutrophils in the cyclophosphamide-treated animals, a majority of these foci were also unable to penetrate past the epidermis. Although Candida yeast proliferated relatively poorly when cultured in homogenates of skin lacking the epidermis, Candida pseudohyphae could invade into the dermis if inoculated skin was isolated from normal animals and cultured in vitro, or if the epidermis was removed by gentle scraping prior to inoculation with Candida yeast onto the remaining skin of leucopenic animals. Therefore, in the absence of neutrophil contact and killing of Candida pseudohyphae in the epidermis, other cutaneous defense mechanisms appear to be capable of preventing invasion of a majority of the organisms into the dermis. These findings may help to explain why deep Candida infections are rare in patients who have extensive superficial candidiasis. PMID- 3045211 TI - Paul Langerhans' death centennial, July 20, 1988. PMID- 3045210 TI - Tumorigenicity and metastatic behavior in nude mice of two human squamous cell carcinoma lines that differ in production of the cytokine ETAF/IL-1. AB - The purpose of these studies was to determine whether the production of the cytokine epidermal cell thymocyte-activating factor (ETAF) by human squamous cell carcinoma (SCC) cells correlated with their tumorigenicity and metastatic potential in athymic nude mice. Cells of the human SCC line A431 produced rapidly growing subcutaneous tumors, few experimental lung metastases, and low levels of ETAF activity in vitro. In contrast, cells of the SCC Colo-16 line produced slower growing subcutaneous tumors, high numbers of experimental lung metastases, and a high level of ETAF activity in culture supernatants. The apparent relationship between production of ETAF and experimental metastasis formation was not consistent. Clonal populations of the SCC A431 and Colo-16 were isolated in vitro. The clones of Colo-16 varied in their ability to produce experimental metastases and in production of ETAF in vitro. However, the levels of ETAF production did not correlate with the propensity of the SCC cells to produce experimental metastases. We conclude that while the growth and metastasis of human SCC in nude mice may benefit from production of the cytokine ETAF, the ETAF production per se is not invariably linked with the capability of the SCC cells to metastasize. PMID- 3045212 TI - Wesley William Spink, 1904-1988. A tribute. PMID- 3045214 TI - Histoplasmosis and AIDS. PMID- 3045213 TI - Etiology and outcome of acute pelvic inflammatory disease. AB - We studied 71 women with the clinical diagnosis of acute pelvic inflammatory disease (PID) by laparoscopy and comprehensive microbiology in order to define the major etiologic determinants of poor fertility prognosis after tubal infection. Fifty women were found to have acute PID. Of the 50 women, 23 were pregnancy seeking and had a subsequent evaluation to determine fertility outcome. Seven of 13 women with non-gonococcal infection had an adverse reproductive outcome, compared with none of 10 women with gonococcal infection (P = .007). Two groups of causes for adverse reproductive outcome were found. Of the seven infertile women, four had initial tubal abscess, and three had evidence of Chlamydia trachomatis infection. This study directly documents the poor fertility prognosis for women with tubal abscess and suggests that women with culture and/or serological evidence of chlamydial infection also have a poor fertility prognosis. PMID- 3045215 TI - A bioemulsifier produced by Candida albicans enhances yeast adherence to intestinal cells. PMID- 3045216 TI - Enteroinvasive Escherichia coli in travelers with diarrhea. PMID- 3045217 TI - Hospital-acquired cryptosporidiosis in a bone marrow transplantation unit. PMID- 3045218 TI - Respiratory syncytial virus pneumonia in a cardiac transplant recipient. PMID- 3045219 TI - Legionellosis in cardiac transplant recipients: results of a nationwide survey. PMID- 3045220 TI - CFA/V is not a new fimbrial type of human colonization factor antigen. PMID- 3045221 TI - Clinical and biological effects of recombinant interferon-beta administered intravenously daily in phase I trial. AB - Interferon-beta serine (IFN-beta ser) was administered intravenously (i.v.) daily for 14 days at doses of 3, 10, 30 X 10(6) units to 19 patients. In this Phase I trial, IFN-beta ser was tolerated without limiting fever or subjective toxicities. At 30 X 10(6) units, 3 patients developed hematologic toxicity and dose escalation was thus terminated. No patient developed detectable binding or neutralizing antibody to IFN-beta. A significant (p less than 0.006) increase in serum beta 2-microglobulin and a significant (less than 0.005) increase in 2',5' oligoadenylate synthetase (2-5A) in peripheral mononuclear cells were identified. Increase in these proteins did not correlate with dose or with the disappearance of serum IFN over the first 5 h after injection. Two patients, one with renal carcinoma and one with melanoma, had objective responses. This trial further confirms safety and biological potency of this synthetic mutant of IFN-beta. PMID- 3045222 TI - Seroepidemiological studies of leprosy in northern Malawi based on an enzyme linked immunosorbent assay using synthetic glycoconjugate antigen. AB - A total of 6002 blood samples from total population samples in four separate areas within Karonga District, Northern Malawi, were tested for anti Mycobacterium leprae antibody using an ELISA based on synthetic glycoconjugate antigen. Results are presented using different criteria for seropositivity. Regardless of the criterion used, the proportion of individuals classified as "positive" rose to a peak at 20-30 years of age and then fell, and it was higher at all ages in females than in males. There was no difference in seropositivity levels between individuals with or without BCG scars. Although leprosy cases, in particular those with positive smears, had higher antibody levels than nonleprosy cases, analysis of age-standardized data revealed only weak evidence for a correlation between the prevalence rates of clinical leprosy and of seropositivity within the four areas. There was no evidence for higher seropositivity levels in household contacts of leprosy cases compared to noncontacts. The implications of these results for the epidemiology of leprosy in this population are discussed. PMID- 3045223 TI - Activity of ofloxacin against Mycobacterium leprae in the mouse. AB - Mice inoculated with 4800 Mycobacterium leprae in the left hind foot pad were treated from day 62 to day 150 after infection with 50 mg or 150 mg of ofloxacin per kg body weight, 150 mg pefloxacin per kg, or 50 mg prothionamide per kg. These drugs were administered by esophageal cannula 5 days weekly with dapsone (0.01 g per 100 g diet). Multiplication of M. leprae in the treated and in untreated control mice was assessed by monthly harvests. The treatment of mice with the smaller dosage ofloxacin, with pefloxacin, prothionamide, or dapsone uniformly resulted in a delay of multiplication of 4 months, compared to the multiplication of M. leprae in the untreated controls. The delay of multiplication (4 months) being 1 month longer than the duration of drug administration (3 months), all of the treatments may be considered as bacteriopausal or moderately bactericidal. In contast with these results, treatment of mice with 150 mg ofloxacin per kg resulted in no growth of the organisms whatever as late as 18 months after inoculation, strongly suggesting that, in that dosage, ofloxacin had killed all of the M. leprae. Such a profound killing activity has been observed only with rifampin. Although the pharmacokinetic characteristics of ofloxacin are different in man from those in the mouse, the daily dosage of 150 mg ofloxacin per kg body weight in the mouse is equivalent to 400 mg per day in man which is the usual therapeutic dosage; thus, the results obtained in the mouse may be extrapolated to man. Therefore, ofloxacin appears a very promising drug for the chemotherapy of leprosy. PMID- 3045224 TI - Identification of T-cell-activating recombinant antigens shared among three candidate antileprosy vaccines, killed M. leprae, M. bovis BCG, and mycobacterium w. AB - Antigenic crossreactivity among three candidate antileprosy vaccines, killed Mycobacterium leprae, BCG, and Mycobacterium w, was studied using T-cell lines and clones raised from BCG- and killed-M. leprae-vaccinated subjects. To identify the crossreactive antigens, the T-cell lines and clones were tested against Escherichia coli lysates containing 65-, 36-, 28-, 18-, and 14-kilodalton (kDa) and 13B3 M. leprae antigens and 65-, 19-, and 12-kDa M. tuberculosis antigens. The short-term T-cell lines, which compared to T-cell clones are easy to raise and maintain, were equally effective in identifying the T-cell-activating recombinant antigens. The reactivity pattern of the T-cell lines and the clones suggested that 65-kDa M. leprae and M. tuberculosis antigens are present in M. leprae, BCG, and Mycobacterium w; 18-kDa M. leprae antigen is shared between M. leprae and Mycobacterium w, 13B3 M. leprae antigen is possessed by M. leprae and BCG. These and other unidentified T-cell-activating antigens shared among candidate leprosy vaccines may be the basis for induction of in vivo sensitization to M. leprae antigens after vaccination with BCG or Mycobacterium w. PMID- 3045225 TI - Partial characterization of antigens from M. leprae evoking IgG and IgM antibodies in armadillos. AB - Armadillo IgG and IgM antibody responses to Mycobacterium leprae were analyzed using isotypic-specific antisera by means of immunoblotting. Blots developed for IgG antibodies to M. leprae showed multiple protein antigens (Mr = 12-90 K) in some heavily infected armadillos. In contrast, blots developed for IgM antibodies to M. leprae showed a single, broad, diffuse band of immunoreactivity at approximately 33 kDa. The 33-kDa immunogen was detectable with silver stain modified for carbohydrate reactivity, suggesting the presence of a polysaccharide component. In addition, binding of 125I-concanavalin A to the 33-kDa component demonstrated the presence of mannose and/or glucose residues. PMID- 3045226 TI - Histologic responses in sixty multibacillary leprosy patients inoculated with autoclaved Mycobacterium leprae and live BCG. AB - Sixty lepromatous or borderline lepromatous patients were submitted to immunotherapy with a mixture of autoclaved Mycobacterium leprae and BCG. The histopathologic findings in skin biopsy specimens taken before and after immunotherapy were evaluated independently by six histopathologists in a workshop setting. Their pooled observations on diagnosis and classification were analyzed to assess the histopathologic changes following various periods of immunotherapy. Expressing the results as the average value of five to six independent observations, there were changes in classification of reversal or upgrading toward the tuberculoid end of the leprosy spectrum in 90.5% of the patients initially classified as lepromatous (LL), and in 83.3% of those initially classified as borderline lepromatous (BL). The histopathologic findings amply support the clinical, bacteriologic and immunological changes following immunotherapy from LL or BL, to BL, mid-borderline (BB) or even borderline tuberculoid (BT) leprosy. PMID- 3045227 TI - Leprosy vaccine--a reappraisal. PMID- 3045228 TI - Leprosy: the immunologist and the patient. PMID- 3045229 TI - Dapsone susceptibility of Mycobacterium leprae isolated before 1977. PMID- 3045230 TI - Perurethral ultrasound-guided ovum pickup. AB - Either a percutaneous-transvesical, a transvaginal, or a perurethral-transvesical approach can be used for oocyte recovery under ultrasound guidance in an in vitro fertilization and embryo transfer program. After having experienced these three different approaches in our program, we preferentially used the perurethral transvesical approach as our routine technique for oocyte recovery under ultrasound guidance. We feel that this method is easier to perform and also carries less risk for contamination. From January to December 1986, 186 oocyte retrievals under ultrasound guidance were performed. In 7 cases no oocytes were found despite normal ovarian stimulation. A total of 767 oocytes was collected; the fertilization rate was 71.8%. Forty pregnancies were achieved (21.5% per attempt or 27.7% per embryo replacement). Except for transient hematuria, no complications were observed. PMID- 3045231 TI - The prediction of local recurrence in rectal adenocarcinoma by histopathological examination. AB - Local recurrence of rectal adenocarcinoma is mainly due to failure to remove all the tumour. A method is described for the routine detection of involvement of the circumferential (lateral) resection margin. Current definitions of the length of the rectum are inadequate for the assessment of the risk of local recurrence as the rectum frequently extends higher than 15 cm. Use of the term recto-sigmoid should be replaced clinically by sigmoidoscopic measurement of the height of a tumour and pathologically by its anatomical relationship to the level of peritoneal reflection, i.e. lower or upper segment of the rectum or the sigmoid colon. Tumours above the peritoneal reflection (upper segment) are at risk of circumferential resection margin involvement due to their retroperitoneal component. The amount of tissue excised varies considerably from surgeon to surgeon. Meticulous attention to the clearance of the tumour at the circumferential resection margin is essential if local recurrence rates are to be reduced. A trial of postoperative radiotherapy should be instigated based on the pathologist's identification of patients at high risk of local recurrence. PMID- 3045232 TI - Respiratory muscle coordination. PMID- 3045233 TI - Platelet adherence to cardiac and noncardiac endothelial cells in culture: lack of a prostacyclin effect. AB - Cardiac tissues show a propensity to develop nonbacterial thrombotic endocarditis, a meshwork of platelets and fibrin. This lesion may cause a predisposition to subsequent colonization by circulating microorganisms, leading to infective endocarditis. We measured platelet adherence in vitro to cultured endothelial cells derived from the porcine aortic valve and ascending aorta. We found that valvular endothelial cells showed a twofold to threefold higher adherence than ascending aortic endothelial cells of chromium 51-labeled platelets in the presence of proteolytically active thrombin. This finding did not correlate with endothelial prostacyclin release: cardiac valve endothelial cells released more prostacyclin than did, ascending aortic cells, exogenous prostacyclin had no effect on thrombin-stimulated adherence, and aspirin inhibition of endothelial prostacyclin synthesis showed no effect on platelet adherence. Fixation of platelets abolished thrombin-stimulated adherence; fixation of endothelial cells had minimal effect. We suggest that these differences may contribute to the propensity of the cardiac valve to develop nonbacterial thrombotic endocarditis. PMID- 3045234 TI - Inhibition of the activation of Hageman factor (factor XII) by platelet factor 4. AB - Platelet factor 4 is a polypeptide constituent of platelet alpha granules that is released during platelet aggregation and inhibits heparin-mediated reactions. Hageman factor (factor XII) is a plasma proenzyme that, when activated by certain negatively charged agents, initiates clotting via the intrinsic pathway of thrombin formation. In earlier studies using crude systems, platelet factor 4 inhibited activation of Hageman factor by dextran sulfate or cerebrosides, but not activation of Hageman factor by kaolin or ellagic acid. In the present study we examined the mechanisms of inhibition by platelet factor 4, using purified reagents. Platelet factor 4 inhibited activation of Hageman factor by ellagic acid, as measured by amidolysis of a synthetic substrate of activated Hageman factor, an effect inhibited by heparin or by an anti-platelet factor 4 antiserum. Coating glass tubes with platelet factor 4 before addition of normal plasma significantly lengthened the partial thromboplastin time of normal plasma. In addition, the clot-promoting properties of kaolin were inhibited by its prior exposure to platelet factor 4. Thus, the inhibitory properties of platelet factor 4 directed against the activation of Hageman factor were confirmed in a purified system. In this purified system, in contrast to earlier studies using crude systems, platelet factor 4 inhibited activation of Hageman factor by glass, ellagic acid, or kaolin. PMID- 3045235 TI - Henry A. Christian (1876-1951). PMID- 3045236 TI - Diagnostic radiology in fertile or pregnant women. PMID- 3045237 TI - Bacterial tracheitis in a young adult. AB - A previously healthy young adult presented with inspiratory stridor and hoarseness but minimal dysphagia. Indirect laryngoscopy and lateral neck X-rays confirmed a diagnosis of membranous tracheitis. This responded to humidification, antibiotics and steroids. Secretions removed at direct laryngoscopy sent for culture grew Staphylococcus aureus. The literature is reviewed. PMID- 3045239 TI - Regulatory and legal issues in the reprocessing and reuse of hemodialyzers. A critical overview. PMID- 3045238 TI - Risk management for hazardous chemicals. OSHA's Hazard Communication Standard and EPA's Emergency Planning and Community Right-to-Know Regulations. PMID- 3045240 TI - Legal implications of computer-aided medical diagnosis. PMID- 3045242 TI - Role of activated macrophages in resistance to systemic candidosis. AB - To evaluate further the contribution of activated macrophages in resistance, the course of systemic candidosis was assayed in beige and NLM mice that had been previously infected with Mycobacterium bovis (BCG). Four weeks following BCG infection, mice were inoculated intravenously with 1 x 10(4) viable Candida albicans. At various times thereafter, the number of C. albicans colony-forming units in the livers, spleens, and kidneys was determined. The average number of CFU recovered from the kidneys of NLM mice decreased throughout the assay and was comparable in both BCG-treated and control mice. In contrast, the number of CFU cultured from the kidneys of untreated control beige mice progressively increased throughout the assay period. This profile of renal susceptibility was not appreciably altered in BCG-treated beige mice. However, fewer (10- to 100-fold) CFU were cultured from the livers and spleens of BCG-treated beige and NLM mice than from untreated controls. These results support the hypothesis that in the absence of functional polymorphonuclear leukocytes, activated macrophages represent a means to control the proliferation of C. albicans. PMID- 3045241 TI - Thymic stroma-derived T-cell growth factor (TSTGF): II. Biochemical and functional characterization. AB - The culture supernatant (SN) from a cloned line of thymic stroma-derived cells in fibroblastic form (TSCF) contained a factor capable of supporting the growth of the interleukin (IL) 2-dependent, antigen-specific helper T cell (Th) clone 9-16 without requiring IL2 and antigen. This active substance, designated as thymic stroma-derived T-cell growth factor (TSTGF), was partially purified through DEAE Sephacel chromatography and PBE 94 chromatofocusing. The original SN did not contain IL1, IL2, IL3, IL4, or interferon activities; but an appreciable magnitude of colony-stimulating factor (CSF) activity in addition to TSTGF was present, whereas the partially purified preparation of TSTGF was depleted of any type of CSF activity. The elution profile of TSTGF activity on the chromatofocusing has revealed that TSTGF has an isoelectric point (pI) of about 6.0. When a purified TSTGF sample was applied to Sephacryl S-200 column chromatography and sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis, TSTGF activity was eluted in a single peak around an apparent molecular weight of about 25,000. The activity of TSTGF also was shown to be relatively stable with heat treatment and in the wide range of pH, but it was abolished by treatment with either trypsin or dithiothreitol. These results indicate that TSTGF, a novel T-cell growth factor, is the protein that has an apparent molecular weight of about 25,000 and a pI of 6.0, and in the intact molecule, it contains the disulfide bond(s) required to maintain and/or express its biologic activity. PMID- 3045243 TI - Computerized tracking for newborn screening and follow-up: a review. AB - In the third decade of newborn screening for phenylketonuria (PKU) and other disorders computers are being used increasingly for both the laboratory and the follow-up aspects of screening programs. In 1984 slightly less than 40% of the state programs had automated follow-up. Lack of funding is probably the major inhibitor of more widespread use of computers in tracking newborns through the newborn screening process. It is suggested that federal funds be made available to ensure wider distribution of currently used tracking systems and development of methods for tracking newborns from birth through follow-up. PMID- 3045244 TI - Role of the adrenal medulla in stress-induced hyperinsulinaemia in normal mice and in mice infected with Bordetella pertussis or treated with pertussis toxin. AB - Intranasal infection of mice with a sublethal dose of Bordetella pertussis or the intravenous administration of purified pertussis toxin resulted in a marked increase in the serum immunoreactive insulin concentration following ether stress. This stress-induced hyperinsulinaemia was not modified significantly by blockade of alpha 2-adrenoceptors with idazoxan, beta-adrenoceptors with propranolol, autonomic ganglia with hexamethonium, opioid receptors with naloxone, muscarinic cholinoceptors with atropine or by adrenal demedullation. The effect of pertussis in promoting stress-induced hyperinsulinaemia was mimicked qualitatively by alpha 2-adrenoceptor blockade, adrenal demedullation or ganglionic blockade. However, the serum immunoreactive insulin response to ether stress was smaller in animals subjected to these procedures compared with the response seen in mice infected with B. pertussis or treated with pertussis toxin. Thus, in the mouse, acute stress produces hyperinsulinaemia under conditions in which the release of adrenal medullary catecholamines is prevented, or the inhibitory action on insulin secretion is blocked by alpha 2-adrenoceptor antagonists or by pertussis toxin. PMID- 3045245 TI - Thyrotrophin-blocking antibodies in goitrous primary hypothyroidism: detection by a highly sensitive bioassay and determination of site of action. AB - Using a highly sensitive bioassay technique, the presence of antibodies capable of blocking the stimulation of thyrocyte function by TSH has been investigated in the sera of a group of 50 patients with primary hypothyroidism. TSH-blocking activity was detected in immunoglobulin (IgG) preparations from the sera of 13 patients (26%). All 13 IgG preparations blocked both TSH-stimulated iodide uptake and cyclic AMP generation (Spearman's rank correlation = 0.6, p = 0.02). However, only one IgG preparation blocked dibutyryl cyclic AMP-stimulated iodide uptake. The presence of TSH-blocking antibody activity was associated with goitrous (ten out of thirteen patients) as well as atrophic (two out of thirteen patients) primary hypothyroidism. Furthermore, TSH-blocking activity was not associated with other thyroid autoantibodies, as thyrotrophin-binding-inhibiting immunoglobulins and anti-TSH antibodies were undetectable in all cases and there was no correlation between TSH-blocking activity and the presence or titre of anti-thyroglobulin or anti-microsomal antibodies. This study indicates that TSH blocking antibodies are present in the serum of some patients with primary hypothyroidism and are directed towards a site, presumably adjacent to or contiguous with the TSH receptor, that is not the binding site for TSH. The coexistence of TSH-blocking activity and goitre in the majority of these patients implies that these antibodies, although capable of blocking TSH-stimulated thyroid hormone biosynthesis, do not necessarily inhibit the mitogenic action of TSH in vivo. PMID- 3045246 TI - The parasympathetic nervous system and glucocorticoid-mediated hyperinsulinaemia in the genetically obese (fa/fa) Zucker rat. AB - Lean (Fa/-) and genetically obese (fa/fa) Zucker rats were adrenalectomized at 18 days of age (3 days before weaning) before the onset of hyperinsulinaemia. At 40 41 days of age, basal and glucose-stimulated insulin concentrations did not differ significantly between lean and obese rats. Plasma insulin and glucose concentrations were higher in both phenotypes 24 h after administration of corticosterone (2.0 mg at 12-h intervals). Corticosterone-treated obese rats had higher basal and glucose-stimulated insulin levels than similarly treated lean animals, although plasma glucose concentrations did not differ between phenotypes. The basal plasma insulin concentration of obese rats treated with corticosterone for 24 h was reduced 15, 30 and 45 min after injection of atropine (0.3 mg) without any significant change in the plasma glucose level. Injection of atropine (0.3 mg) 20 min before a glucose load prevented the greater increment in plasma insulin concentration of corticosterone-treated obese rats compared with similarly treated lean animals. Atropine administration (0.3 mg) to intact obese rats at 40 days of age reduced, but did not abolish, their hyperinsulinaemia compared with intact lean animals. It is concluded that (1) pre-weaning adrenalectomy prevents the development of hyperinsulinaemia in genetically obese rats, (2) corticosterone replacement for only 24 h restores the hyperinsulinaemia of obese rats, (3) the differential effects of corticosterone on insulin secretion by lean and obese rats are mediated by the parasympathetic nervous system and (4) the parasympathetic nervous system contributes to, but is not the only cause of, hyperinsulinaemia in intact obese rats. PMID- 3045247 TI - Mutational hot spots in Ig V region genes of human follicular lymphomas. AB - The genes coding for the Ig light chains expressed in two cases of human follicular lymphoma were cloned and sequenced. In each case, multiple independent isolates of the tumor population were compared. Although each tumor represented a single clone of B cells with a unique V/J joint, different cells within each tumor had accumulated multiple point mutations in the V gene during clonal expansion. Most of the mutations observed were silent, but some resulted in amino acid replacements. Identical silent mutations were often observed in independent isolates of each tumor. By combining the current data with VH sequences obtained previously from the same cells, it was apparent that the repetitive silent mutations could not be explained solely by a genealogic tree. Such mutations could represent hot spots whose tendency to mutate may be influenced by neighboring DNA sequences or by the methylation of specific cytosine residues. PMID- 3045249 TI - Effects of T cell depletion in radiation bone marrow chimeras. II. Requirement for allogeneic T cells in the reconstituting bone marrow inoculum for subsequent resistance to breaking of tolerance. AB - The ability of normal recipient-type lymphocytes to break tolerance in long-term allogenic radiation chimeras has been investigated. Reconstitution of lethally irradiated mice with a mixture of syngeneic and allogeneic T cell-depleted (TCD) bone marrow (BM) has previously been shown to lead to mixed chimerism and permanent, specific tolerance to donor and host alloantigen (3-5). If allogeneic T cells are not depleted from the reconstituting inoculum, complete allogeneic chimerism results; however, no clinical evidence for GVHD is observed, presumably due to the protective effect provided by syngeneic TCD BM. This model has now been used to study the effects of allogenic T cells administered in reconstituting BM inocula on stability of long-term tolerance. We have attempted to break tolerance in long-term chimeras originally reconstituted with TCD or non TCD BM by challenging them with inocula containing normal, nontolerant recipient strain lymphocytes. tolerance was broken with remarkable ease in recipients of mixed marrow inocula in which both original BM components were TCD. In contrast, tolerance in chimeras originally reconstituted with non-TCD allogeneic BM was not affected by such inocula. Susceptibility to loss of chimerism and tolerance was not related to initial levels of chimerism per se, but rather to T cell depletion of allogeneic BM, since chimeras reconstituted with TCD allogeneic BM alone (mean level of allogeneic chimerism 98%) were as susceptible as mixed chimeras to the tolerance-breaking effects of such inocula. The possible contribution of GVH reactivity to this resistance was investigated using an F1 into parent strain combination. In these animals, the use of non-TCD F1 BM inocula for reconstitution did not lead to resistance to the tolerance-breaking effects of recipient strain splenocytes. These results suggest that the ability of T cells in allogeneic BM inocula to confer resistance to late graft failure may be related to their graft-vs.-host reactivity, even in situations in which they do not cause clinical GVHD. These findings may have relevance to the mechanism whereby T cell depletion of allogeneic BM leads to an increased incidence of late graft failure in clinical BM transplantation situations. PMID- 3045248 TI - Amastigotes of Trypanosoma cruzi sustain an infective cycle in mammalian cells. AB - The two main stages of development of the protozoan parasite Trypanosoma cruzi found in the vertebrate host are the trypomastigote and the amastigote. It has been generally assumed that only trypomastigotes are capable of entering cells and that amastigotes are the intracellular replicative form of the parasite. We show here that after incubation for 4 h with human monocytes in vitro 90% or more of extracellularly derived (24 h) amastigotes of T. cruzi are taken up by the cells. Within 2 h they escape the phagocytic vacuole and enter the cytoplasm, where they divide and after 4-5 d transform into trypomastigotes. Trypomastigotes also invade cultured human monocytes. However, they show a lag of several hours between invasion and the start of DNA duplication, while amastigotes commence replication without an apparent lag. Amastigotes also infect cultured fibroblasts, albeit with lower efficiency. When injected intraperitoneally into mice, amastigotes are as infective as trypomastigotes. Based on these results, and on prior findings that amastigotes are found free in the circulation of mice during the acute stage of the disease (3), it seems likely that the cellular uptake of amastigotes can initiate an alternative subcycle within the life cycle of this parasite in the mammalian host. Also, because trypomastigotes and amastigotes have diverse surface antigens, they may use different strategies to invade host cells. PMID- 3045251 TI - Long-term residence of a graft is an insufficient stimulus for the induction of tolerance. Investigating the role of cyclosporine in class I-disparate heart grafts in the rat. AB - This study demonstrates that the induction of tolerance is possible across a class I only antigenic barrier that fails to produce heart graft rejection. However, the long-term residence alone of such a graft per se, does not necessarily lead to the establishment of systemic tolerance in the recipient. The important finding in this study with regard to the biology of allograft tolerance, is that while the class I antigen provides the stimulus, its presence alone is not sufficient for the induction of tolerance; indeed, the action of the Cyclosporine A (CyA) is a necessary adjunct to its induction. PMID- 3045252 TI - Respiratory syncytial virus: heterogeneity of subgroup B strains. AB - In order to investigate further possible structural differences among the two subgroups of respiratory syncytial virus (RSV), we analysed the antigenic characteristics and size of structural proteins of 20 subgroup A and 43 subgroup B strains by their reactions with monoclonal antibodies (MAbs) directed against the proteins of RSV using immunofluorescence, ELISA and radioimmunoprecipitation assays. The latter test also enabled determination of the size of different structural components. The 37 MAbs employed were generated by immunization with both subgroup A and B strains. They represented specificities for distinct epitopes on five different structural proteins. The subgroup A strains proved to be relatively uniform. The fusion (F) protein, nucleoprotein (NP) and matrix (M) proteins of all strains tested had the same Mr and all except one strain had a phosphoprotein (P protein) of the same Mr. The F and P proteins were lower in Mr in B strains compared to A strains, which confirmed previous findings. The Mr of the large surface glycoprotein (G protein) of subgroup A strains varied slightly, probably on the basis of differing glycosylation. By contrast, the subgroup B strains exhibited substantial variation in the Mr of the G and also the P proteins and in reactivity with MAbs directed against the G and F proteins. Three size classes of the P protein were identified in B strains: 33K to 34K, 32K to 33K, and 31K to 32K. Twenty-seven subgroup B strains failed to react with four anti-G MAbs representing a single epitope, G2; the remaining 16 strains reacted with these MAbs. We designated these two sets of variants of B strains B1, which lacked the epitope, and B2, which had the epitope. The B1 strains also varied in the size of the G and P proteins. In contrast, all B2 strains had large G proteins and all except two strains had relatively large P proteins (33K to 34K). All subgroup B1 and B2 strains exhibited the same sizes of NP, F and M proteins. We conclude that the subgroup B strains of RSV include two variants, B1 and B2, and that the major difference between them resides in the G and P proteins. PMID- 3045250 TI - Normal mouse peritoneum contains a large population of Ly-1+ (CD5) B cells that recognize phosphatidyl choline. Relationship to cells that secrete hemolytic antibody specific for autologous erythrocytes. AB - We have found that, in the peritoneums of normal adult mice, 5-15% of lymphocytes bind a fluorescent liposome probe. In ontogeny, cells with this specificity were shown to appear by 8 d after birth, and increase to the adult frequency by 2-3 wk. Some older mice contain an expanded population of these cells. We have shown that liposome binding occurs by cell surface IgM recognizing the common membrane phospholipid, phosphatidyl choline (PtC). Virtually all of these PtC-specific cells bear the cell surface marker Ly-1. Our results indicate that roughly 1 in 10 peritoneal Ly-1+ B cells has this single specificity. We have found that the precursors to all the cells that form plaques on protease-treated autologous erythrocytes (BrMRBC) are included in the PtC-specific population and can be isolated by FACS. We believe this is the first report of sorting large numbers of B cells with a single antigen specificity from normal, unimmunized animals. This method will allow for in vitro and in vivo studies of differentiative and proliferative properties of Ly-1+ B cells, which may help define their role in development and disease. PMID- 3045253 TI - Multiplication of influenza A viruses with cleavable and non-cleavable haemagglutinin in chicken embryo membranes or organs, and cell cultures derived therefrom. AB - Pathogenic properties of influenza A viruses introduced into embryonated chicken eggs via the allantoic cavity, the amniotic cavity or the yolk sac were studied using viruses with cleavable or non-cleavable haemagglutinin (HA), or reassortants derived from the highly pathogenic fowl plague virus (FPV) which has a cleavable HA. The various organs, membranes and fluids were analysed for virus yields, and by immunohistochemistry for production of viral nucleoprotein. Virus replication in primary tissue cultures derived from various embryonic organs was also studied. Some of the reassortants, which were previously found to be non pathogenic for chickens and were temperature-sensitive (ts) at 40 degrees C, multiplied at 37 degrees C to the same extent as the highly pathogenic FPV. The spread of other non-pathogenic reassortants in the embryo was restricted. For example, 81/Ho was able to multiply to a reasonable extent only in the chorioallantoic and the allantoamniotic membranes. After inoculation of the A/PR/8/34 strain containing a non-cleavable HA into the amniotic cavity, virus multipled only in the inner epithelial layer of the amnion, in superficial epidermal cells and in superficial epithelia of the oropharyngeal cavities, the nasal and paranasal sinuses and the oesophagus. Kidneys were free of virus antigen, although the virus multiplied to high titres in primary tissue cultures derived from embryonic kidneys. Thus influenza A viruses can be non-pathogenic for chickens because they are ts and unable to multiply at the body temperature of chickens (41 degrees C), or because their spread in the animal is impaired per se. PMID- 3045254 TI - Characterization of potentially foetotropic Palyam serogroup orbiviruses isolated in Zimbabwe. AB - Twelve Palyam serogroup orbiviruses isolated in Zimbabwe from aborted cattle foetuses, plus one isolated from the visceral organs of a cow and another from vulture faeces, were examined in comparison with known members of the serogroup by complement fixation, indirect immunofluorescence, fluorescent focus reduction neutralization tests and PAGE of the segmented, dsRNA genomes. The viruses were indistinguishable from known members of the serogroup by complement fixation and indirect immunofluorescence, but two novel viruses, for which the names Gweru and Marondera are proposed, and two previously described viruses, Nyabira and Abadina, were identified by neutralization tests. The dsRNA profiles of Abadina serotype isolates differed from that of the Abadina prototype virus, indicating that different electropherotypes may occur within serotypes. PMID- 3045256 TI - [Bourneville's tuberous sclerosis]. AB - Among the group of hereditary histodysplasia, Bourneville's tuberous sclerosis demonstrate original and important place. Its clinical and histopathological polymorphism make more difficult the diagnosis because many symptoms are non specific and/or appeared at different age of life. The variability of the expression and of the penetrance are a very serious unpeachement for the genetic counselling. A recent reevaluation of several series of case reports seems to demonstrate that the frequency of new mutations has been probably surestimated. The gene location in 9q3-4 is quite certain and will induce soon the possibility of a more efficient prenatal diagnosis. The gene action mechanism at the embryonic development is probably correlated with the "oncogene character" of the specific mutation. PMID- 3045257 TI - [Steinert's disease (dystrophia myotonica). General review]. AB - By comments of 9 tables the main clinical and genetic features of myotonic dystrophy are recalled. Due to a most variable penetrance and expressivity, the recognition of the adult form of this autosomal dominant disease can be difficult. Congenital myotonic dystrophy is a serious disease which represents a major genetic risk for the heterozygous women, which are often very slightly affected or even not aware of their disorder. For them, prenatal diagnosis is fully justified and now possible with a small risk of error by determination of DNA polymorphism. Preclinical detection in young adults can also be improved by this new method. PMID- 3045255 TI - DNA-binding proteins induced by the cottontail rabbit herpesvirus CTHV. AB - Several new polypeptides were detected in cells infected with CTHV, a cottontail rabbit herpesvirus. All of them (Mr 150K, 110K, 93K, 83K, 75K and 35K) accumulated in the nucleus during the infectious cycle, and all except the 150K species bound to DNA-cellulose affinity columns in low-salt buffers. Polyclonal antisera prepared against the 35K DNA-binding protein also recognized the 75K species. Although the 75K protein could be detected earlier in infection than the 35K protein, late in the infectious cycle the latter increased to an abundance approaching that of cellular histones. Treatment of partially purified virions with a non-ionic detergent indicated that the 35K protein, but not the 75K protein, is a component of capsid/tegument structures. The anti-35K/75K serum did not cross-react with herpesvirus sylvilagus virion proteins, which, in an electrophoretic comparison, exhibited both similarities to and differences from the virion proteins of CTHV. Labelling of CTHV-infected cells with [32P]orthophosphate revealed the presence of phosphoproteins electrophoretically comigrating with the 93K, 83K, 75K and 35K proteins. PMID- 3045258 TI - HBeAg/anti-HBe seroconversion during and after protracted immunosuppressive treatment in type B chronic hepatitis. AB - Seventy-seven consecutive HBeAg-positive chronic hepatitis patients were studied from 1971 to 1983 to establish the seroconversion rate in the e system. Patients with less than a year of follow-up were not included in the study. Fifty-six patients with chronic active hepatitis (CAH) received immunosuppressive treatment (corticosteroids combined with azathioprine). The remaining twenty-one patients received no treatment, nine of them with chronic persistent hepatitis (CPH) and 12 with CAH. A retrospective study was performed with stored sera samples: HBeAg and anti-HBe were determined by RIA, and results were correlated with alanine aminotransferase (ALAT) levels in the same samples. The linearized seroconversion rate from HBeAg to anti-HBe was expressed as percent per patient-year. It was 9.6% in CPH patients and 8.8% in CAH patients without treatment. In CAH patients under immunosuppressive drugs it was as low as 1.1% and increased to 28.7% when treatment was withdrawn. ALAT levels were significantly lower in total seroconverted patients when compared with nonseroconverted (NS) patients, but no difference was found between partial seroconverted (PS) and NS patients. The results suggest that although immunosuppressive drug withdrawal may enhance seroconversion rate in type B CAH, delayed seroconversion and reported side effects during treatment stand against protracted usage of these drugs. PMID- 3045259 TI - Intelligent computer-based assessment and psychotherapy. An expert system for sexual dysfunction. AB - New and converging developments in the areas of artificial intelligence, intelligent tutoring systems, and cognitive therapy have made possible a new approach to computer-assisted assessment and psychotherapy. This new approach combines the capacity for intelligent therapeutic dialogue with the presentation of individualized therapeutic interventions. Previous attempts at computerized psychotherapy are reviewed to highlight a newly developed rule-based expert system, Sexpert, which assesses and treats sexual dysfunction. Preliminary observations concerning couples' reaction to and acceptance of Sexpert are presented. PMID- 3045260 TI - Methadone maintenance to abstinence. How many make it? AB - This study imposed a set of decision rules to review and compare methadone maintenance detoxification research results in three 5-year eras since 1970. Variables of interest were detoxification completion rates, relative completion rates with and without program review and approval, psychotherapy, new pharmacological agents to accelerate the detoxification process, and abstinence rates at follow-up as a function of such treatment variables. The review found progressive improvement in overall detoxification completion rates over the three 5-year eras: 39.7%, 54.9%, and 76.3%. Program-recommended detoxification showed much higher completion rates than did unrecommended detoxification in 1970-1975 and psychotherapy-assisted detoxification showed modest but consistently greater completion rates in the two eras during which it was studied, 1970-1975 and 1976 1980. The consistent gains in detoxification completion rates over the three eras are attributed primarily to the use of new drugs which greatly shorten the detoxification interval and ameliorate withdrawal symptoms. The drugs do not appear to have had an effect on follow-up abstinence rates. Limitations upon the conclusions and generalizability of findings are discussed. Suggestions are made for future methadone maintenance detoxification treatment and research. PMID- 3045262 TI - Ozone air quality models. A critical review. PMID- 3045261 TI - Functional properties of the subtype of insulin receptor found on neurons. AB - In this report, we have examined the structure, regulation, and function of insulin receptors in cultured neurons from fetal chicken brain. The apparent molecular weight of the alpha-subunit of neuronal insulin receptors, analyzed by photoaffinity labeling and sodium dodecyl sulfate gel electrophoresis under reducing conditions, was 115,000. The number of insulin receptors in the cultures increased from day 2 to day 4 during a period of extensive process formation. After 5 days in culture, there were approximately 40,000 high-affinity insulin receptors per neuron. When neurons were photoaffinity labeled at 16 degrees C and then warmed to 37 degrees C for 30 min, approximately 40% of the cell-surface receptors were recovered in the intracellular, trypsin-insensitive pool. Chronic exposure of neurons to insulin (100 ng/ml) resulted in a time-dependent loss of neuronal insulin receptors with a maximal decrease of 50% after 24 h. Insulin had no effect on glucose transport, glucose oxidation, or glycogen synthase activity in neurons. On the other hand, insulin supported the growth and differentiation of a fraction of neurons isolated from chick forebrain. We conclude that (1) cultured neurons from fetal chicken brain express the same subtype of insulin receptor previously identified in adult rat and human brain, (2) the neuronal subtype of insulin receptor undergoes internalization and down-regulation in response to insulin, and (3) neuronal insulin receptors do not acutely regulate glucose metabolism but mediate growth in neurons. PMID- 3045263 TI - Improved tumor localization with increasing dose of indium-111-labeled anti carcinoembryonic antigen monoclonal antibody ZCE-025 in metastatic colorectal cancer. AB - Monoclonal antibodies (MoAbs) against carcinoembryonic antigen (CEA) react with human colorectal cancer cells, and when labeled with a gamma-emitting radioisotope, may help to localize known and occult metastatic disease. We tested ZCE-025 (Hybritech, Inc, San Diego), a high-affinity immune gamma globulin1 (IgG1) MoAb anti-CEA that does not react with normal granulocyte glycoproteins in a phase I/II trial to determine the reagent's toxicity and its maximum efficacy in detecting metastatic colorectal cancer. Increasing doses of unlabeled ZCE-025 were mixed with 1 mg of Indium-111 (111In)-radiolabeled MoAb and administered intravenously (IV) to 34 patients who had metastatic colorectal cancer. Planar nuclear or single photon emission computed tomographic (SPECT) scans were performed 48 to 72 and 120 to 144 hours later. Total dose of MoAb and scanning sensitivity (number of imaged lesions/number of known lesions) were correlated up to 80 mg. At doses of 2.5 to 20 mg, a mean of 22% of the lesions were imaged; at 40 mg, 77% were imaged (P less than .01). Liver metastases were detected as areas of increased activity ("hot") at the 40 mg dose but showed decreased MoAb uptake at lower doses. At the 40 mg dose normal liver parenchymal uptake of the labeled MoAb was lower with respect to blood pool compared with the other doses. At 80 mg, however, sensitivity of detection declined to 21%. One milligram of 111In labeled ZCE-025 antibody coinfused with 39 mg of unlabeled antibody appeared optimal for detecting metastatic colorectal cancer, particularly in the liver. Although the exact mechanism(s) for this dose effect is currently unknown, a partial "blocking" effect of unlabeled antibody with a change in MoAb biodistribution may be occurring. PMID- 3045264 TI - Early stage ovarian cancer: a randomized clinical trial comparing whole abdominal radiotherapy, melphalan, and intraperitoneal chromic phosphate: a National Cancer Institute of Canada Clinical Trials Group report. AB - Two hundred fifty-seven eligible patients with stage I, IIA "high risk" ovarian carcinoma and IIB, IIIO (disease confined to pelvis), were randomized to either total abdominal radiotherapy (arm A) 2,250 rad in 20 fractions (107 patients), melphalan (arm B) 8 mg/m2/d X 4 every 4 weeks X 18 courses (106 patients), or intraperitoneal chromic phosphate (arm C) 10 to 20 mCi (44 patients). All patients were initially treated with pelvic radiotherapy; arm A, 2,250 rad in ten fractions; and arms B and C, 4,500 rad in 20 fractions. Entry to arm C was discontinued early because of toxicity. In a multifactor analysis using proportional hazards models, no significant difference in survival was observed although there was a marginally significant difference in disease-free survival (P = .015) with arm B being superior to arm A. Stage (P less than .0001), grade (P less than .0001), and histology (P less than .008) were predictors of survival in the multifactor analysis. Performance status, age, and residual disease were significant predictors in the single factor analysis but were not predictive when correction was made for the effects of stage, grade, and histology. Five-year survival rates are 62% for arm A, 61% for arm B, and 66% for arm C. Median duration of follow-up is 8 years. Long-term complications of radiotherapy were seen in 19 patients on arm A, 11 on arm B, and 11 on arm C. Four patients who had received melphalan developed either a myelodysplastic syndrome or acute leukemia. Violations in covering the whole abdominal target volume were correlated with survival. PMID- 3045265 TI - Autologous bone marrow transplantation for patients with poor-prognosis lymphoma. AB - Review of prognostic factors at Memorial Hospital in New York City has shown that adult patients with large-cell lymphoma (diffuse histiocytic lymphoma by Rappaport classification) who have high lactic dehydrogenase (LDH) and/or bulky mediastinal or abdominal disease are destined to do poorly with conventional combination chemotherapy, with a 2-year disease-free survival of about 20%. Patients who relapse after conventional combination chemotherapy have a similar poor prognosis. Thirty-one such patients with lymphoma were studied to evaluate the efficacy of intensive radiotherapy (hyperfractionated total body irradiation [TBI] [1,320 rad]), and cyclophosphamide (60 mg/kg/d for two days) followed by autologous bone marrow transplantation (ABMT). Our results show a disease-free survival advantage (P = .002) for 14 patients who underwent ABMT immediately after induction of remission with 79% surviving at a median follow-up 49.2+ months, compared with a median survival of 5.2 months for 17 patients administered ABMT while in relapse and/or after failing conventional treatment. Our results support the use of aggressive therapy as early treatment for patients with poor prognostic features. PMID- 3045266 TI - High-dose combination alkylating agent therapy with autologous bone marrow rescue for refractory solid tumors. AB - Twenty-six adults, ages 27 to 60, with refractory metastatic solid tumors were treated with high-dose cyclophosphamide (Cy) + carmustine (BCNU) at one of three escalating dose schedules followed by autologous bone marrow transplantation (ABMT). Toxicity was severe and dose-related, with the maximum tolerated dose for the combination determined to be Cy 160 mg/kg and BCNU 900 mg/m2. Median time to WBC recovery (greater than or equal to 1,000/microL) was 13 days post-ABMT (range, nine to 22 days) and to a platelet count of greater than or equal to 50,000/microL, 22 days (range, 13 to 83 days). Sixteen of 20 evaluable patients (80%) responded to therapy with at least 50% reduction in measurable tumor, and three patients achieved complete remission (CR). Responders included eight of nine evaluable patients with breast carcinoma, two of five with melanoma, two of two with sarcoma, and four of four with colon carcinoma. Response durations were short (median, 4 months), even for complete responders, and relapses generally occurred at sites of previous metastases. In order for this approach to have a more significant impact on overall survival, it may need to be applied earlier in the natural history of the malignancy. PMID- 3045267 TI - Interpretation of clinical trials in diffuse large-cell lymphoma. AB - Diffuse large-cell lymphoma is one of the neoplasms curable with chemotherapy in an appreciable percentage of patients. However, all patients are not cured and the best combination of agents is not certain. This reflects the lack of completed comparative trials using the regimens that appear most effective. Despite this uncertainty, several principles for the therapy of diffuse large cell lymphoma can be identified that allow an analysis of the results reported in the literature. These principles include the following: (1) for a regimen to be curative in a substantial number of patients it must achieve a high rate of complete remissions; (2) cure must be accomplished with frontline therapy; (3) drugs must be delivered at curative doses; (4) rapidity of achieving a complete response might be related to chance for cure; (5) prolonged treatment for diffuse large-cell lymphoma is unnecessary; and (6) aggressive therapy is toxic. In analyzing the results with any regimen it is important to have long follow-up since late relapses do occur and initial very positive results tend to decay with greater numbers of patients treated. Applying these principles to the reported chemotherapy studies in patients with diffuse large-cell lymphoma suggest that no one of the new regimens is clearly superior to the others. Also, it is not clear that cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP) delivered at full doses to comparable patients is inferior to the newer regimens. The results of ongoing studies comparing these regimens might help resolve these questions. PMID- 3045268 TI - Perioperative blood transfusions and cancer recurrence. AB - This collective review addresses the issue of transfusion-induced immunosuppression as it relates to patients undergoing cancer surgery. Patients receiving perioperative blood transfusions have a significantly worse prognosis than patients undergoing cancer surgery without a perioperative transfusion. It is thought that this is because transfusions produce a nonspecific immunosuppression by increasing the number and/or activity of suppressor T lymphocytes, decreasing the number of natural killer cells, and inducing anti idiotypic antibodies. This risk, particularly when considered with the other risks of transfusion such as hepatitis or the acquired immunodeficiency syndrome (AIDS), suggests that criteria for transfusion of these patients should be stringent and related to an unequivocal need for replenishment of RBCs. PMID- 3045269 TI - MAP2 is a component of crossbridges between microtubules and neurofilaments in the neuronal cytoskeleton: quick-freeze, deep-etch immunoelectron microscopy and reconstitution studies. AB - Microtubules (MT) and neurofilaments (NF) are linked by frequent crossbridges in situ. In order to answer the question of what makes these crossbridges, we performed the immunogold procedure on rat spinal cord motor neurons using an affinity-purified polyclonal antibody against rat brain MAP2 and gold-labeled anti-rabbit IgG goat IgG. A quick-freeze, deep-etch technique (QF-DE) in conjunction with decoration with anti-MAP2 antibody and ferritin-labeled second antibody was also used. In motor neuron dendrites crossbridges were clearly displayed between MTs and NFs by QF-DE. These crossbridges were revealed in thin sections as fuzzy filamentous structures between MT and NF. Gold particles studded the fuzzy structures associated with MT. Many such structures connected MTs to NFs. Furthermore, antibody complexes containing ferritin were localized on the crossbridges between MTs and NFs by the QF-DE study. In addition, we performed reconstitution experiments. We isolated 70 kDa (L) protein of neurofilaments from calf spinal cords and assembled L to form neurofilaments in vitro. MAP2 bound these neurofilaments according to both SDS-PAGE and QF-DE electron microscopy of the pellets of suspensions containing L proteins and MAP2. When we added tubulin to this suspension and polymerized it in the presence of taxol, neurofilaments were crosslinked with microtubules by MAP2 crossbridges. Hence, from these 2 approaches we concluded that MAP2 is a component of crossbridges between MTs and NFs in the neuronal cytoskeleton in vivo and in vitro. PMID- 3045270 TI - Cultured smooth muscle targets lack survival activity for ciliary ganglion neurons. AB - Cultured neurons require specific trophic agents in order to survive. This dependence is thought to resemble the neuron-target interdependence that develops in vivo during synaptogenesis and neuronal cell death. The notion that neurons in general derive trophic support from their synaptic targets is based primarily on studies of peripheral neurons and motor neurons. To assess the general applicability of this nerve-target relationship, we tested the ability of vascular smooth muscle (VSM) to support dissociated neurons from the chick ciliary ganglion. The ciliary ganglion contains 2 distinct neuronal populations, one of which innervates striated muscle, the other VSM. Striated muscle cocultures are known to support all of the neurons in the ganglion for extended periods. Dissociated neurons were therefore cocultured in microwells containing VSM derived from the rat or chick aorta and from the choroid coat of the chick eye. Surviving neurons were counted after 1, 2, 5, and 7 d. Striated muscle is able to support full neuronal survival in the same assay. However, in no case was VSM capable of contributing to neuronal survival in vitro. The neurons in the VSM cocultures were able to form neurites and make contacts with their putative targets, as confirmed by scanning electron and light microscopy. The presence of viable and differentiated smooth muscle cells was demonstrated in the cultures by transmission electron microscopy and analysis of smooth muscle alpha-actin. The failure of VSM and even the choroid target tissue to support the survival of their innervating neurons suggests that novel mechanisms may operate to provide trophic support for neurons innervating VSM targets. PMID- 3045271 TI - Value of myocardial defect size measured by thallium-201 SPECT: results of a multicenter trial comparing heparin and a new fibrinolytic agent. AB - In a multicenter randomized double-blind trial comparing heparin and a new fibrinolytic agent, anisoylated plasminogen streptokinase activator complex (APSAC), 231 patients presenting with a less than 5 hr acute myocardial infarction underwent a contrast angiography (CA) before the end of the first week of admission, and radionuclide cardiac blood-pool imaging and a 201Tl single photon emission computed tomography (SPECT) study before the end of the third week. Left ventricular ejection fraction (LVEF) and a wall motion score (WM) were calculated from CA. LVEF was also obtained from cardiac blood-pool imaging, and defect size (DS) from 201Tl SPECT. Results demonstrated that all parameters were significantly improved in patients treated with APSAC versus heparin (contrast LVEF 53 +/- 13 vs. 47 +/- 14 p less than 0.01, WM 9.8 +/- 6.5 vs. 13.3 +/- 7.9 p less than 0.001, radionuclide LVEF 43 +/- 12 vs. 40 +/- 13 p less than 0.05, DS 14 +/- 12 vs. 18 +/- 14 p less than 0.05). When the patients were divided according to infarct site and infarct-related coronary artery patency, it was demonstrated with all four parameters that the beneficial effect of APSAC can be largely explained by the lower incidence of vessel obstruction in this group (37% vs. 77% in the heparin group, p less than 0.001). It is concluded that (a) when compared with heparin and in the conditions of the trial, APSAC significantly improves the cardiac function and decreases the DS and (b) DS measured by 201Tl SPECT is as valuable a quantitative parameter of therapeutic evaluation as are LVEF and WM. PMID- 3045272 TI - Intravascular angiomatosis (Masson's lesion). AB - Intravascular angiomatosis, also known as Massons' lesion, is a relatively uncommon, although important, tumor because it clinically mimics such benign lesions as mucocele, pyogenic granuloma, and hemangioma, as well as malignant neoplasms such as angiosarcoma and Kaposi's sarcoma. More important is the potential for misdiagnosis at the histological level that poses a significant problem to those unfamiliar with this lesion. This article reviews the literature on perioral intravascular angiomatosis and presents five additional cases. PMID- 3045273 TI - Erythematous gingival enlargement in diabetic patients: a report of four cases. PMID- 3045274 TI - Synovial hemangioma of the temporomandibular joint: report of a case and review of the literature. AB - This is the first report of a localized intra-articular synovial hemangioma of the temporomandibular joint. Although, a rare finding, synovial hemangioma should be included in the differential diagnosis of soft tissue lesions in this region. PMID- 3045275 TI - Direct bonding of arch bars in the management of maxillomandibular injuries. AB - The shortcomings of the conventional arch bars for the treatment of maxillomandibular injuries are described. A mesh-backed arch bar bonded to the teeth is recommended as a means of overcoming these problems. PMID- 3045277 TI - Solid cell nests of the thyroid. AB - The ultimobranchial thyroid solid cell nests (SCN), irregular structures of about 1 mm in maximal diameter, are usually found in the middle third of the thyroid lateral lobes. SCN are basically composed of non-keratinizing epidermoid cells which lack intercellular bridges and are immunohistochemically positive for a panel of high and low molecular weight keratin proteins, as well as for carcinoembryonic antigen. In addition, SCN display isolated or grouped peripheral calcitonin-immunoreactive 'clear' (C) cells in up to 54 per cent of cases. The SCN central lumen, when present, is usually surrounded by mucinous cells; in addition, it may contain desquamated cells, cell debris, acid mucosubstances, characteristic PAS-positive granular material after diastase treatment, and colloid-like material. The so-called mixed follicles, structures lined by epidermoid cells of SCN and follicular epithelium, are often found as an additional component of the ultimobranchial remnants. The relationship of SCN to thyroid parenchymal cells and the probable implications of the thyroid 'ultimobranchial system' to tumour histogenesis are analysed. Pitfalls that may emerge with regard to SCN in practical pathological approaches are emphasized. PMID- 3045276 TI - Optimal preservation of p21 ras immunoreactivity and morphology in paraffin embedded tissue. AB - Specific immunostaining of p21 ras protein by the well-characterized pan-ras antibody Y13-259 is achieved in paraffin sections of human and animal tissues fixed in periodate-lysine-paraformaldehyde-dichromate (PLPD). Intensity of staining is as good as in cryostat sections, with superior histological detail. Localization to plasma membrane is demonstrated in rodent cells genetically manipulated to express abundant p21 ras (the FHO5T1 cell line), both in preparations suspended in agar after culture in vitro and in those growing as tumour in vivo. Strong positive staining is observed in neoplasms of human breast and colon, tissues in which there is independent evidence of elevated ras gene expression. The superior morphology afforded by this technique allows clear characterization of p21 ras expression in small premalignant lesions for which other methods of detection of oncogene expression are not appropriate. PMID- 3045278 TI - Axillary lymph node status in carcinoma of the breast. AB - In this prospective study, axillary clearance specimens from consecutive routine mastectomies and wide excisions for carcinoma were divided into three segments by sutures in the theatre: apical (furthest from the tumour), mid and basal (nearest to the tumour) segments. Each specimen was then randomized to either formalin or Carnoy's fixation. Following fixation, lymph nodes were dissected from each of the three segments, sectioned once through the hilum, and examined microscopically. There was a significant (P less than 0.05) difference in the total number and number of reactive lymph nodes harvested from Carnoy's compared with formalin-fixed material. The number of nodes containing metastatic carcinoma was not significantly different (P = 0.64). In several cases, involved apical segment lymph nodes were found with no involvement of the proximal groups; this observation may have serious implications for the procedure of proximal lymph node sampling in the surgical management of breast cancer. It is suggested that clearing of axillary contents with Carnoy's fixative does not provide any additional information of clinical significance, although it may allow the pathologist to identify lymph nodes rapidly. PMID- 3045279 TI - Citrobacter meningitis and brain abscess in infancy: epidemiology, pathogenesis, and treatment. PMID- 3045280 TI - Avoiding phenylketonuria: why parents seek prenatal diagnosis. PMID- 3045281 TI - Clinical trial of naloxone in birth asphyxia. AB - To determine whether endogenous opiates play a role in the pathogenesis of perinatal asphyxia, a blinded clinical trial of naloxone, a competitive opiate receptor blocker, was undertaken in infants with low 1-minute Apgar scores. Of 85 infants with 1-minute Apgar score 0 to 3, 44 received an injection of naloxone (approximately 0.4 mg/kg) and 41 received saline solution. In 108 infants with 1 minute Apgar score 4 to 6, 54 received naloxone and 54 saline solution. In neither group was there a significant effect of naloxone on respiratory frequency or heart rate up to 30 minutes after injection, nor at 24 hours of age. In both groups active muscle tone of upper and lower limbs was increased by naloxone, a response that may not be beneficial in the face of inadequate oxygen delivery to vital organs. We conclude that naloxone at this dose had no readily apparent benefit in the resuscitation of the asphyxiated newborn infant. PMID- 3045283 TI - Recurrent abdominal pain and ascites in an adolescent. PMID- 3045282 TI - Renal calcification in preterm infants: pathophysiology and long-term sequelae. AB - We examined the clinical course of 17 preterm infants with chronic lung disease who received loop diuretics and developed nephrocalcinosis; nine of them were followed for up to 4.5 years. The mean gestational age was 26.8 weeks (SD 2.2 weeks), and mean birth weight was 830 gm (SD 276 gm). The diagnosis of renal calcification was made at a mean age of 12 weeks (SD 6.5 weeks) by abdominal x ray examination, screening abdominal ultrasound studies or, both. Calcification was associated with both furosemide therapy and the presence of multiple potential risk factors. Renal calcification, length, and function were subsequently evaluated in nine patients at a mean age of 21.3 months (SD 15.3 months). Improvement in calcification occurred in five patients, with total resolution in four. Renal length, determined by ultrasound examination and corrected for body length, was normal in 17 of 18 kidneys. Serum creatinine values and calculated glomerular filtration rates were abnormal in four of nine patients. We conclude that renal calcification in preterm infants is associated with multiple risk factors, including furosemide usage, and tends to diminish during the first year of life. However, renal function may remain compromised in some patients. PMID- 3045284 TI - Festschrift in honor of Amos Christie, M.D. PMID- 3045285 TI - Ceftazidime versus placebo in respiratory exacerbations of cystic fibrosis. PMID- 3045286 TI - Thyroid ultrasonography in congenital hypothyroidism. PMID- 3045287 TI - Adjustable suture strabismus surgery for acquired vertical deviations. AB - Forty-seven patients undergoing 51 adjustable suture strabismus procedures for acquired vertical deviations were evaluated for preoperative and adjustment factors which might influence the postoperative alignment. The type of deviation, surgical procedure, previous surgery, immediate postadjustment alignment, and immediate postadjustment versions were assessed. Oblique muscle surgery in addition to an adjustable vertical rectus muscle significantly decreased the success rate (p = .0303). Other less important factors were a previous history of strabismus surgery and a moderate limitation of versions following the adjustment. A slight over- or undercorrection after adjustment did not affect the success. PMID- 3045288 TI - Vertical effect of the adjustable Harada-Ito procedure. AB - The adjustable Harada-Ito procedure, as described by Metz, was performed on four patients. A hypotropia of 10 to 12 prism diopters (PD), in addition to incyclotorsion, was induced in two patients in whom the superior oblique tendon was split 8 mm, only enough to permit mobilization to its new location. Splitting the tendon for 15 mm eliminated the induced hypotropia while preserving incyclotorsion. PMID- 3045289 TI - Familial Brown's syndrome. AB - We present findings in two siblings with bilateral Brown's syndrome and review other reported familial cases. PMID- 3045290 TI - A rationale for monitoring the periodontal microbiota after periodontal treatment. AB - Strict control of dental plaque accumulation remains the cornerstone of successful maintenance of the treated patient. However, some recurrences of periodontitis appear to take place despite good oral hygiene habits. A proportion of these are likely due to qualitative features of the periodontal microbiota rather than merely increased mass. Microscopic as well as cultural monitoring of qualitative features of the periodontal microbiota may be helpful in detecting critical changes. This review presents some of the research findings that provide a rationale for microbiological monitoring. Some of the advantages as well as difficulties encountered with these procedures are discussed. PMID- 3045291 TI - Acute nonlymphocytic leukemia, monocytic variant. Report of a case. AB - This report documents a case of a patient presenting unusual oral features of one of the rarer variants of acute nonlymphocytic leukemia, a malignancy whose oral manifestations may be the first indication of the presence of an underlying blood dyscrasia. The findings at the initial appointment and the patient's progress for the following 13 months until his death are presented. Initial suspicion of the clinical symptoms and an immediate, subsequent hospital hematology report established the diagnosis. The histologic features of a gingival biopsy taken during the oral examination were strongly suggestive of leukemia. Since the leukemias may imitate other oral conditions, especially various diseases of the periodontium, it is of paramount importance that the dental clinician recognize their clinical manifestations. As future treatment modalities improve, the dental clinician is seeing more leukemic patients in remission as well as more cases that have relapsed. PMID- 3045293 TI - Effect of sodium intake on urinary renin excretion in rats. AB - The present study was carried out to investigate the effect of sodium intake on urinary renin excretion in rats. Renin activity was measured before and after acidification, by radioimmunoassay of angiotensin I generated by incubation with partially purified homologous renin substrate. The molecular weight of renin was determined by gel filtration. Low sodium intake for 4 weeks resulted in a 16-fold increase in urinary renin excretion and led to a 15-fold increase in plasma renin activity. A 2.5-fold increase in renal renin content was simultaneously observed. In contrast, high sodium intake for 4 weeks decreased urinary renin excretion, plasma renin activity and renal renin content to about 20%, 25% and 35% of the control level, respectively. No significant change in renin activity after acidification was observed in all the urine samples. The molecular weight of renin in the urine from rats on low or high sodium intake was 40,000, the value being identical with that from control rats. These results indicate that urinary renin excretion is well correlated with plasma renin activity, and that only active renin with a molecular weight of 40,000 is excreted in the urine of rats, even under conditions of altered sodium intake. PMID- 3045292 TI - [Pyrogens and endotoxins. I. History, origins, sources and properties]. PMID- 3045294 TI - Effect of lipid composition on insulin-mediated fusion of small unilamellar liposomes: a kinetic study. AB - Liposome-entrapped insulin could be used to prolong the hypoglycemic action of insulin. Also, the conjugation of insulin to the surface of liposomes allows the potential application of using insulin as a transporting molecule to deliver liposome-entrapped drugs to insulin-receptor rich tissues. The success of these two approaches of drug delivery depends on how insulin may interact with liposomes. The present study describes the application of the principle of kinetics to investigate the effect of insulin on the stability of various preparations of liposomal drug carriers. The technique of fluorescence resonance energy transfer was employed to study the destabilization of liposomal formulations through the process of insulin-mediated fusion of liposomes. The kinetics of insulin-mediated fusion appeared to be compatible with a model whereby the initial rate of fusion is governed by the mechanism of fusion of two small unilamellar, unfused liposomal particles. The rate constants of insulin mediated fusion of various liposomal formulations were estimated from the initial rate of fusion, using the model of two-particle fusion. Arrhenius analysis of the rate constants of fusion at different temperatures suggests that the mechanism of insulin-mediated fusion of small unilamellar vesicles is not governed merely by the energy and frequency of collision between liposomal particles. Other factors, such as the binding of insulin with the surface of liposomes and the temperature effect on the dynamics of the liposomal membrane, as well as conformation of insulin, could potentially be important.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3045296 TI - [Eicosanoid cascade in the gastrointestinal tract]. PMID- 3045297 TI - Evolution of psychiatric community care. PMID- 3045298 TI - Lupus: early history of the wolf. PMID- 3045295 TI - Effects of interrupting the renin-angiotensin system on sodium excretion in man. AB - 1. Twelve normal volunteers were studied on 2 separate days, having taken a range of diets providing 50-300 mmol sodium per day for 3 days and having been dehydrated overnight. Each volunteer received a background intravenous infusion of arginine vasopressin (5 x 10(-7) i.u. kg-1 min-1) on both days, and also received 6 mg captopril orally on one day and a placebo tablet on the other. The ensuing changes in arterial pressure, and in urinary solute and solute-free water excretion were recorded. 2. Captopril did not significantly alter arterial pressure. It increased the rate of excretion of sodium but not of potassium, and it did not significantly change urinary osmolality or creatinine clearance. 3. Captopril increased the rate of solute-free water reabsorption and did so in direct proportion to its effect of increasing sodium excretion. 4. A further twelve normal, dehydrated volunteers on free diets were studied on each of 2 days, after taking 500 mg lithium carbonate on the previous evening. On each day, they also received a loading dose and maintenance infusion of inulin. On one day they received 50 mg captopril orally, and, on the other, they received a placebo tablet. The arterial pressure, urinary excretion of electrolytes, and inulin clearance were recorded. 5. Captopril increased the rates of excretion of sodium, lithium and potassium, but there were no significant changes in inulin clearance or arterial pressure. 6. The natriuretic effect of captopril in either group of twelve volunteers was not significantly less in those volunteers who were already excreting more sodium, at least over the range of dietary sodium loading to which the volunteers were subjected. 7. Six normal volunteers were studied on a further 2 days, having taken a diet supplying 30 mmol sodium per day for 3 days and being dehydrated overnight. Each volunteer received a background intravenous infusion of arginine vasopressin (5 x 10(-7) i.u. kg-1 min-1) on both days and also received an intravenous infusion of saralasin acetate (50 ng kg-1 min-1) plus carrier on one day and carrier alone on the other. The ensuing changes in arterial pressure, and in urinary solute and solute-free water excretion were recorded. 8. There was a small but significant fall in systolic arterial pressure during the infusion of saralasin.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3045299 TI - Hieronymus Fracastorius 1483-1553. PMID- 3045300 TI - The viable child. The Croonian lecture 1988. PMID- 3045301 TI - To keepe good dyet... PMID- 3045302 TI - It's magic! PMID- 3045303 TI - Factors influencing dental decision making. AB - In clinical decision making, dentists routinely choose between alternative treatments such as crown vs amalgam/composite buildup, root canal vs extraction, fixed bridge vs removable partial denture, and prophylaxis vs subgingival curettage or periodontal scaling. A number of technical and patient factors can influence dentists' choice of treatment in these situations; however, little is known about their relative importance. To address this issue, a list of technical (e.g., periodontal status and caries rate) and patient (e.g., cost and patient preference) factors possibly influencing choice of treatment was developed for each pair of services. Responding to a mail questionnaire, 156 general dentists in Washington State listed the top three factors influencing their choice of service in each pair. Results revealed that dentists took different factors into account in choosing among alternative treatments. Technical factors dominated over patient concerns; only about 33 percent of the dentists considered patient factors important in choosing alternative therapies. The latter group was less preventively oriented, were solo practitioners, worked longer hours, and had lower prices. Results suggest patients may have little influence on prescriptions of therapy among experienced general dentists. PMID- 3045304 TI - Risk assessment of mercury exposure from dental amalgams. AB - Much attention is being focused upon the issue of mercury exposure from dental amalgam restorations and the potential for adverse health effects. This controversy has grown beyond the confines of the dental profession itself and is becoming an emotional public health issue. In hope of regaining good health, many dental patients with chronic systemic diseases are considering replacement of their amalgams. Dentists are increasingly being challenged to prove the safety of amalgams. Recently, systematic methods have been established for quantitative evaluation of environmental risks. This study brings together the quantitative methodologies of risk assessment and the knowledge of mercury exposure from dental amalgams to estimate the safety of dental amalgam restorative therapy. Analysis concludes that the margin of safety for mercury toxicity in humans from dental amalgams is approximately 8- to 30-fold. There are many uncertainties involved in this estimate, and further studies are warranted to improve its precision. PMID- 3045305 TI - The John W. Knutson Distinguished Service Award in Dental Public Health--1987 recipient: Martha Jane Howard Fales. PMID- 3045306 TI - Pattern of growth of dominant follicles during the oestrous cycle of heifers. AB - Ovarian follicular development was studied in 13 heifers by daily ultrasound examination during 2 complete and consecutive natural oestrous cycles. In 21 cycles (81%) 3 dominant follicles were identified, in 4 cycles (15%) 2 and in the remaining cycle 1 (4%). Consistently, the first dominant follicle was detected on average on Day 4, reached a maximum size on Day 6, went through a period of relative stability between Days 6 and 10, then began to decrease in size and was undetectable by Day 15. The second dominant follicle was detected by Day 12, reached maximum size on Day 16 (or 19 in the 4 cycles in which the 2nd dominant follicle was the ovulatory follicle) and was undetectable by Day 19. The 3rd (ovulatory) follicle was identified on average by Day 16 (range Days 10 to 19) and maximum size was reached on Day 21. The ovulatory follicles were larger (P less than 0.05) than the previous ones and the stage of the cycle at which maximum size was reached was significantly different for each dominant follicle (P less than 0.05). The analysis of the rates of growth and atresia suggest that the rate of growth is slowest during mid-cycle. The number of dominant follicles that developed in the ovary ipsilateral to the corpus luteum was greater (P less than 0.05) than in the contralateral ovary. PMID- 3045307 TI - Exogenous GnRH induces ovulation in seasonally anoestrous lactating goats (Capra hircus). AB - A specific sheep LH radioimmunoassay was validated for the measurement of goat LH, and used to monitor luteal-phase LH episodes and the preavulatory LH surge in progestagen sponge-synchronized cycling goats. No luteal-phase LH episodes were detected during 12 h of frequent (15-min) blood sampling in 2 goats. A preovulatory LH surge was recorded in 5/5 goats, with a mean amplitude of 45.4 +/ 7.2 ng/ml and a mean time of onset of 38.4 +/- 1.2 h after removal of a progestagen-impregnated sponge. In anoestrous goats, single i.v. injections of 1000 and 2000 ng GnRH induced LH episodes with a mean amplitude of 2.04 +/- 0.11 and 3.67 +/- 0.06 ng/ml respectively, but injections of 250 or 500 ng did not consistently elevate LH concentrations. Progestagen-primed, seasonally anoestrous lactating goats were treated with repeated injections of 1500 ng GnRH (every 2 h for 52 or 78 h) in May 1985 or 1986. All 10 had kidded in March of the same year, and were consequently at peak lactation at the time of GnRH treatment. A preovulatory LH surge was detected in 9 goats with a mean time of onset of 59.5 +/- 2.9 h (1985) or 39.6 +/- 3.3 h (1986) after vaginal sponge removal. All animals displayed oestrus and ovulated, and 9 of the goats were mated: in 5 of these animals pregnancies were successfully carried to term. The results show episodic LH release in response to GnRH and indicate that ovulation can be induced in seasonally anoestrous goats, even at peak lactation, and normal pregnancies may result. PMID- 3045308 TI - Reproduction in a laboratory colony of the pouched mouse, Saccostomus campestris. AB - Pouched mice ovulate spontaneously and have a 4-day cycle (3-5 days). The variation was caused by prolonged oestrus. The vagina opened at about 34 days of age and the first oestrus was experienced at 44 days of age. Females experienced several sterile cycles before their first conception, which occurred at an age of about 56 days. The gestation period in most cases was 21 days and implantation occurred about 4-5 days after mating. The females were not receptive post partum. Litter size varied from 3 to 13 with a mean of 7.1 in young primiparous females and 7.9 in adult multiparous females, indicating that fecundity did not increase with age or parity in this species. Mortality of young was highest during the first 2 days post partum. The young were not attached to the nipples and were weaned at 25 days of age. Females did not cycle during lactation. After lactation most females exhibited one or two oestrous cycles without mating or became oestrous and mated without conceiving, resulting in a litter interval of about 53 days. PMID- 3045309 TI - A PMSG/GnRH method for the superovulation of the monovulatory brush-tailed possum (Trichosurus vulpecula). AB - Optimal ovarian stimulation with minimal degenerative changes was achieved in brush-tailed possums 3 days after a subcutaneous injection of 10 i.u. PMSG. This treatment alone did not result in ovulation and only rarely in oocyte maturation. Ovulation of 8-24 mature oocytes occurred when on the 3rd day after 10 i.u. PMSG the female received 3 intramuscular injections of 50 micrograms synthetic gonadotrophin-releasing hormone (GnRH) 90 min apart. Superovulation was achieved in immature, cycling, pregnant and lactating females, but not in animals in the preovulatory phase of a natural oestrous cycle. PMID- 3045311 TI - Havelock Ellis. PMID- 3045310 TI - Feverfew--an ancient remedy for modern times? PMID- 3045312 TI - The chemical death of Lot's wife: discussion paper. PMID- 3045313 TI - Pseudoachalasia of the cardia: a review. PMID- 3045314 TI - Vitreous fluorophotometry: a review. PMID- 3045316 TI - Havelock Ellis: his literary and political activities as a medical student at St Thomas' Hospital London, 1881-1887. PMID- 3045315 TI - Cardiac pacing in the accident and emergency department: a review. PMID- 3045317 TI - Paul Langerhans (1847-1888): a centenary tribute. PMID- 3045318 TI - Yersinia pseudotuberculosis ileitis presenting with severe intestinal haemorrhage. PMID- 3045319 TI - An analysis of current methodologies for conformational searching of complex molecules. PMID- 3045320 TI - Design of rat renin inhibitory peptides. AB - Because several well-studied strains of rats manifest spontaneous hypertension, we set out to design a renin inhibitor suitable for use in this species. On the basis of the sequence of the renin substrate, a series of substrate analogue inhibitory peptides were synthesized by systematically modifying the P5, P3, P2, P1P1', P2', P3', and P4' positions. In assays against rat plasma renin, we found that modifications at the C-terminal segment have a marked influence on potency, and that a secondary butyl side chain at the P2' position is important for obtaining optimal activity. The structure at the P3' position, however, could vary considerably without significant effect. The steric effect of the P2 position was important; there an isopropyl side chain provided optimal binding between the inhibitor and the enzyme. At the P3 and P5 positions, potency appeared to depend on aromatic side chains. The effects at the P1P1' position of the transition-state residue (3S,4S)-4-amino-3-hydroxy -6-methylheptanoic acid (statine) and its congeners (3S,4S)-4-amino-3-hydroxy-5-phenylpentanoic acid (AHPPA) and (3S,4S)-4-amino-3-hydroxy-5-cyclohexylpentanoic acid (ACHPA) were found to depend on the sequence of the C-terminal segment. For peptides with an unfavorable C-terminal segment (-Ile-Phe-NH2), AHPPA and ACHPA resulted in a surprising retention of potency. For peptides with a favorable C-terminal segment (-Leu-Phe-NH2), the effect of AHPPA was mild, even though ACHPA still significantly enhanced potency. The hypotensive and plasma renin inhibitory effects of three of the analogues were then studied in anesthetized sodium depleted rats. One of the compounds, acetyl-His-Pro-Phe-Val-Statine-Leu-Phe-NH2 (IC50 against rat plasma renin of 30 nM at pH 7.4), proved to be a potent hypotensive agent and a potentially useful probe for the study of the renin angiotensin system in rats. PMID- 3045321 TI - Synthesis and evaluation of multisubstrate inhibitors of an oncogene-encoded tyrosine-specific protein kinase. 2. AB - Tyrosine-specific protein kinases that transfer the terminal phosphate from ATP to protein acceptors are associated with certain transforming viruses and cell surface growth factor receptors. Here we describe the synthesis and testing of potential multisubstrate inhibitors of this class of enzymes. The inhibitors were prepared by covalent attachment of the terminal phosphate of ATP or its tetraphosphate analogue to tyrosine mimics. Testing against p60v-abl, the tyrosine kinase from the Abelson murine leukemia virus, showed that the series of inhibitors was moderately potent (IC50 values as low as 13 microM). However, structural modification of the tyrosine mimic, including replacement with a serine-like moiety, had little effect on potency. It is therefore concluded that the ATP moiety is largely responsible for binding and that the enzyme requires additional structural features for recognition of the tyrosine-containing substrate. PMID- 3045322 TI - Improved algorithms for automatic bone histomorphometry on a numerized image analysis system. AB - Quantitative histological methods have proved to be the most effective methods in bone disease research. Faster and more accurate techniques are currently needed. We have developed a simple digitized image analysis system which allows accurate measurements of trabecular bone mass. The algorithm is based on the 'four connected sets' mathematical theory. Given a numerized image displayed by a CCD camera, the algorithm recognizes all possible four-connected sets and provides area measurements. The first procedure automatically eliminates small, irrelevant profiles (wrinkles, cell nuclei, etc.) while larger profiles are erased interactively. The second procedure similarly erases the artefactual defects within the trabeculae (artefactual cracks or empty osteocytic lacunae). The method was shown to be very accurate and time-saving. PMID- 3045323 TI - Classification of images of biomolecular assemblies: a study of ribosomes and ribosomal subunits of Escherichia coli. AB - Images of macromolecules obtained in the electron microscope are subjected to correspondence analysis. The structure inherent in the data in the resulting low dimensional factor space is characterized by a mixed classification method which combines the dynamic clouds clustering technique with hierarchical ascendant classification (HAC). For our data, the rejection of marginal clusters obtained by dynamic clouds clustering appears as a crucial prerequisite for a stable performance of HAC. The method is applied to two sets of 204 and 177 images that show the 70S ribosome of Escherichia coli, in the range of overlap views as defined by A. Verschoor and co-workers, and to two sets of 480 and 496 images of the 50S subunit of E. coli depleted of L7/L12 proteins in the well-defined crown view. Reproducible classes are obtained, which are characterized by images reconstituted from factorial coordinates. These classes appear to be related to different orientations on the specimen grid (in the case of the 70S particle) and to different conformational states (50S subunit). PMID- 3045325 TI - Selection of DNA binding sites by regulatory proteins. II. The binding specificity of cyclic AMP receptor protein to recognition sites. AB - The statistics of base-pair usage within known recognition sites for a particular DNA-binding protein can be used to estimate the relative protein binding affinities to these sites, as well as to sites containing any other combinations of base-pairs. As has been described elsewhere, the connection between base-pair statistics and binding free energy is made by an equal probability selection assumption; i.e. that all base-pair sequences that provide appropriate binding strength are equally likely to have been chosen as recognition sites in the course of evolution. This is analogous to a statistical-mechanical system where all configurations with the same energy are equally likely to occur. In this communication, we apply the statistical-mechanical selection theory to analyze the base-pair statistics of the known recognition sequences for the cyclic AMP receptor protein (CRP). The theoretical predictions are found to be in reasonable agreement with binding data for those sequences for which experimental binding information is available, thus lending support to the basic assumptions of the selection theory. On the basis of this agreement, we can predict the affinity for CRP binding to any base-pair sequence, albeit with a large statistical uncertainty. When the known recognition sites for CRP are ranked according to predicted binding affinities, we find that the ranking is consistent with the hypothesis that the level of function of these sites parallels their fractional saturation with CRP-cAMP under in-vivo conditions. When applied to the entire genome, the theory predicts the existence of a large number of randomly occurring "pseudosites" with strong binding affinity for CRP. It appears that most CRP molecules are engaged in non-productive binding at non-specific or pseudospecific sites under in-vivo conditions. In this sense, the specificity of the CRP binding site is very low. Relative specificity requirements for polymerases, repressors and activators are compared in light of the results of this and the first paper in this series. PMID- 3045326 TI - Interactions between Escherichia coli RNA polymerase and lambda repressor. Mutations in PRM affect repression of PR. AB - The rightward operator, OR, of bacteriophage lambda is part of a complex regulatory region that includes PRM, the promoter for repressor synthesis by a prophage, the rightward early promoter PR, and three repressor-binding sites, OR1, OR2 and OR3. By binding to OR2, repressor blocks transcription from PR and simultaneously stimulates the formation of open complexes between RNA polymerase and PRM. In this letter, we describe a test of the hypothesis that the interaction between RNA polymerase bound at PRM and repressor bound at OR2 increases the apparent affinity of repressor for OR. One implication of this hypothesis is that the amount of repressor required for repression of PR should be inversely correlated with PRM promoter strength. This is indeed the case. The amount of repressor required for 50% repression of PR is decreased by prmup-1, an "up" mutation of PRM, and is increased by prm- mutations. An unexpected finding is that in addition to their effect on the apparent affinity of repressor for OR, mutations in the -35 region of PRM alter the shape of repressor-titration curves. We propose that these mutations alter the interaction between RNA polymerase bound at PRM and repressor bound at OR2 in such a way that cooperativity in the binding of repressor to OR1 and OR2 is also disrupted. PMID- 3045324 TI - Spinach chloroplast rpoBC genes encode three subunits of the chloroplast RNA polymerase. AB - Sequence analysis of a 12,400 base-pair region of the spinach chloroplast genome indicates the presence of three genes encoding subunits of the chloroplast RNA polymerase. These genes are analogous to the rpoBC operon of Escherichia coli, with some significant differences. The first gene, termed rpoB, encodes a 121,000 Mr homologue of the bacterial beta subunit. The second and third genes, termed rpoC1 and rpoC2, encode 78,000 and 154,000 Mr proteins homologous to the N and C terminal portions, respectively, of the bacterial beta' subunit. RNA mapping analysis indicates that the three genes are cotranscribed, and that a single intron occurs in the rpoC1 gene. No splicing occurs within the rpoC2 gene or between rpoC1 and rpoC2. Furthermore, the data indicate the possibility of an alternative splice acceptor site for the rpoC1 intron that would give rise to a 71,000 Mr gene product. Thus, with the inclusion of the alpha subunit encoded by rpoA at a separate locus, the chloroplast genome is predicted to encode four subunits (respectively called alpha, beta, beta', beta") equivalent to the three subunits of the core enzyme of the E. coli RNA polymerase. PMID- 3045327 TI - Crystallization of O6-methylguanine-DNA methyltransferase from Escherichia coli. AB - The 19,000 Mr C-terminal domain of the Escherichia coli ada gene product that contains O6-methylguanine-DNA methyltransferase DNA repair activity has been crystallized in a low-salt environment. The crystals, which diffract to 2.3 A (1 A = 0.1 nm), are suitable for detailed structural studies. The space group is P21 with unit cell dimensions a = 46.3 A, b = 45.8 A, c = 46.9 A and beta = 113.3 degrees. PMID- 3045328 TI - Preliminary X-ray crystallographic study of adenylosuccinate synthetase from Escherichia coli. AB - Adenylosuccinate synthetase, a dimeric enzyme of 96,000 Mr, catalyzes the first committed step toward the de novo biosynthesis of AMP. Large, single crystals of adenylosuccinate synthetase from Escherichia coli grow from solutions of polyethylene glycol and ammonium sulfate. Crystals from ammonium sulfate belong to the orthorhombic space group P212121 with unit cell parameters a = 79.0 A, b = 70.2 A and c = 152.6 A. Crystals from polyethylene glycol belong to the space group P21, having unit cell parameters of a = 71.16 A, b = 71.99 A, c = 82.95 A, and beta = 71.52 degrees. The asymmetric units of both crystal forms probably contain the entire dimeric enzyme. PMID- 3045329 TI - Role of toxic effects of oxygen in reperfusion damage. PMID- 3045331 TI - Lipid burden in ischemic myocardium. PMID- 3045330 TI - The role of xanthine oxidase during myocardial ischemia in several species including man. PMID- 3045333 TI - Medical response to radiation and nuclear accidents: lessons for the future. PMID- 3045332 TI - Of clinical alerts and peer review. PMID- 3045334 TI - Human natural lymphocyte effector cells: definition, analysis of activity, and clinical effectiveness. AB - Lymphokine-activated killing has enormous potential in the treatment of cancer. Lymphoid effectors have the potential to recognize and lyse a wide array of tumor cells, a phenomenon which has been seen in vitro and to some extent in vivo. However, studies have indicated that complexity exists not only in the nature of the lymphocyte that can be activated ex vivo for therapeutic use, but also in the delivery of such therapy in a clinical setting. This review attempts to deal with both issues. The nature of the cells mediating lymphokine-activated killer activity, their heterogeneous phenotype, their activation requirements, and a hypothetical mechanism of action are discussed. In addition, previous clinical studies are reviewed and key issues are raised that need to be addressed in upcoming clinical trials. PMID- 3045335 TI - Cephalotaxine esters: antileukemic advance or therapeutic failure? AB - Clinical trials conducted in the People's Republic of China and the United States of the antileukemic efficacy of the cephalotaxine esters are reviewed. Harrington has been incorporated into combination regimens for the treatment of newly diagnosed acute nonlymphocytic leukemia (ANLL) in China, and activity with cephalotaxine esters has also been noted in chronic myelogenous leukemia. While the investigational agent homoharringtonine has shown some activity in the United States in ANLL, investigator interest in the United States has waned because of toxicity and inconvenient schedules. The Chinese trials have used different schedules than have U.S. studies and have been associated with less toxicity. These trials provide new information that may lead to further investigations of the cephalotaxine esters in the United States. PMID- 3045336 TI - Mutagenicity of urine from greenhouse workers. AB - The mutagenicity of urine from spray applicators in 12 greenhouses in 1986 has been evaluated. The workers served as their own controls. Urine samples reflecting pesticide exposure were collected at the end of the day of application and a corresponding control sample was collected 3 d later. Using Salmonella typhimurium bacterial tester strains TA98 and TA100 with and without S9 activation, seven workers showed no significant differences (p less than 0.05) in the mutagenicity of their exposed and control urine. Of the five remaining workers, three, who wore no respirators, showed significantly higher (p less than 0.05) concentrations of mutagens in their exposed urine sample as compared to their respective controls. The mutagenicity of certain of the compounds applied by these latter workers is discussed. PMID- 3045337 TI - Early diagnosis of invasive candidiasis and rapid evaluation of antifungal therapy by combined use of conventional chromogenic limulus test and a newly developed endotoxin specific assay. AB - Since beta-1,3-glucan is a common component of fungal cell wall, its detection might be useful in diagnosing invasive candidiasis. Not only endotoxin but beta 1,3-glucan activates proclotting enzyme contained in a conventional chromogenic limulus test (CCLT). Endotoxin activates this enzyme through factor C, while the beta-1,3-glucan activates through factor G. Since endotoxin specific test (EST) contains factor C, endotoxin would be quantified. By subtracting EST value from CCLT value, beta-1,3-glucan would be quantified. We named this value Fungal Index (FI), and examined if it actually reflects the candidal infection. Ninety-two patients were tested for CCLT and EST prospectively. FI increased significantly in candidal infection (p less than 0.05) but remained low in GNR infection. Moreover, FI increased proportionally to the severity of candidal infection. Elevated FI decreased when antifungal therapy was successful. Thus FI was a useful index not only in the diagnosis of invasive candidiasis but also in the evaluation of antifungal therapy. PMID- 3045338 TI - Artificial intelligence: a computerized decision aid for trauma. AB - A computerized decision support system has been developed to advise ATLS-trained surgeons on the initial definitive management of patients with penetrating injuries of the abdomen immediately following resuscitation and stabilization. The program was developed as an "expert system," using the techniques of artificial intelligence. It is able to suggest: the need for further examination; additional tests; diagnoses; and treatments. In this study, the advice offered by the expert system was compared to that of physicians-in-training. Five actual patient care situations were presented to the system and to 13 medical students and surgical residents: four MS-III, three PGY-I, three PGY-III, and three PGY-V. The suggestions of each of the 13 trainees, the advice of the expert system, and the actual management were blinded. Five surgeons versed in trauma and otherwise not involved in the project judged whether each of the 15 purported management plans was acceptable and ranked them in order of preference. Only the actual care and the advice from the system were judged acceptable for all five problems. The rankings of the expert system were better than those of any individual trainee. The differences were statistically significant for two of the three chief residents, five of nine residents overall, and all four students. This preliminary validation of a prototype expert system is encouraging for the prospect of a computerized decision support system that can help surgeons make initial definitive management plans for patients with major trauma. PMID- 3045339 TI - Medical management of nephrolithiasis in Dallas: update 1987. AB - With available medical treatment programs a remission of stone disease could be achieved in more than 80 per cent of the patients and a decrease in individual stone formation rate obtained in greater than 90 per cent. The need for stone removal may be reduced dramatically by an effective prophylactic program. There is some evidence that certain stones (even calcareous types) may undergo dissolution in vivo with appropriate therapy. Moreover, properly applied medical treatments may be capable of overcoming nonrenal manifestations as well as preventing new stone formation. Thus, the potential development of bone disease in patients with renal tubular acidosis may be averted by potassium citrate therapy. Despite these advantages it is clear that the medical treatment approach cannot provide total control of the disease. Stone disease generally presents with a surgical problem related to an already formed stone before medical diagnosis and selective treatment may be applied. Some patients, albeit a minority, are recalcitrant to medical treatment no matter how heroic. A satisfactory response to medical treatment requires continued compliance by the patient to the recommended treatment program and a commitment by the physician to provide long-term followup care. There is no cure, only prophylaxis. The increasing ease and decreasing cost of new approaches to stone removal, particularly with the advent of second generation extracorporeal lithotripsy, will undoubtedly cast a continuing uncertainty on the need for medical diagnosis and treatment. Several factors might influence the choice between surgical and medical approaches. One factor is the severity of stone disease. Patients with repeated episodes of stone formation might be more likely to adopt preventive therapy, whereas those with infrequent stone episodes may elect simply to have them removed upon their occurrence without medical treatment between episodes. Also, the possibility that lithotripsy may cause long-term hazards (for example development of hypertension) must be clarified. Another factor is the occurrence of extrarenal manifestations. In patients suffering from systemic disorders in which nephrolithiasis is only 1 manifestation (for example distal renal tubular acidosis) a medical approach may be justified exclusive of effects on stone formation. Finally, one must consider the relative practicality and cost between stone removal and a medical approach. It is likely that improvements and reductions in costs will occur with both approaches. It is hoped that urologists and internists work jointly to find an appropriate balance between the 2 approaches.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3045340 TI - Anal submucosal injection: a new route for drug administration in pelvic malignancies. II. Methotrexate anal injection in the treatment of advanced bladder cancer. Preliminary study. AB - The clinical efficacy of submucosal anal injections of methotrexate in advanced bladder cancer is investigated. An experimental study on 20 mice has shown that methotrexate injected into the anal submucosa has no clinicopathological effect on the anorectum. The clinical study comprised 18 patients with advanced bladder cancer (13 with stage T3 and 5 with stage T4 disease) as a test group in whom methotrexate was injected into the anal submucosa and 8 (6 with stage T3 and 2 with stage T4 cancer) treated concurrently with intravenous methotrexate. The dose in both groups was 50 mg. every 5 days for 5 consecutive doses. The course was repeated at 3-week intervals. Most patients received methotrexate as outpatients. Methotrexate blood levels were measured 4 and 24 hours after administration in both groups. In the test group 10 of the 18 patients showed complete tumor regression and were alive 21 to 50 months after the start of treatment. Partial regression was observed in 8 patients. Hematological reserve remained unchanged. Mild toxicity occurred in 3 patients. Of the 8 patients treated intravenously the tumor showed partial regression in 1, was stable in 3 and progressed in 4. Side effects were severe in 5 patients. Our results show that methotrexate injection is highly effective in the treatment of advanced bladder cancer. It is safe, well tolerated and can be used on an outpatient basis. PMID- 3045341 TI - Efficiency and side effects of prostaglandin E1 in the treatment of erectile dysfunction. AB - The efficiency and side effects of a single dose of intracorporeally applied prostaglandin E1 (20 mcg.) in inducing penile erection were examined. In addition, the effect of this dose of prostaglandin E1 was compared to the effect of 7.5 mg. papaverine plus 0.25 mg. phentolamine in a double-blind, crossover designed study. We tested twice 12 men 52.9 +/- 7.6 years old (mean +/- standard deviation) with erectile dysfunction. On 1 occasion the subjects received 20 mcg. prostaglandin E1 and on the other they received 7.5 mg. papaverine and 0.25 mg. phentolamine. At this dosage prostaglandin E1 was most effective in inducing artificial penile erection (11 of 12 patients). However, 75 per cent of the subjects reported burning sensations during the entire period of erection and in 1 prostaglandin E1 treatment resulted in a sustained erection. At the doses used, prostaglandin E1 was more effective in inducing penile erection than papaverine plus phentolamine (11 versus 6 patients). Intracavernous injection of prostaglandin E1 is a potent tool for artificial penile erection and warrants precise examination for its potential clinical use. PMID- 3045342 TI - Local antibody in semen for rapid diagnosis of Chlamydia trachomatis epididymitis. AB - Etiologic studies including micro-immunofluorescence serology for Chlamydia trachomatis were done on 45 consecutive men with acute epididymitis. Of the men 21, all less than 35 years old, had type specific Chlamydia trachomatis antibody in the semen. All patients with semen antibody also had Chlamydia trachomatis antibody in the serum, while only a few of the patients without semen antibody had serum antibody. Chlamydia antibody titers in the semen specimens were higher than those in the sera and they persisted longer. In only 1 patient with semen antibody was another potential etiological agent for epididymitis demonstrated, while most of the patients without semen antibody had bacterial causes for the epididymitis. It was concluded that measurement of Chlamydia trachomatis antibody in semen offered a noninvasive, sensitive and specific method, useful despite prior antibiotic therapy, for diagnosis of the etiology of epididymitis in young men. PMID- 3045344 TI - Endourethroplasty in the management of complicated posterior urethral strictures. AB - A 2-step endourethroplasty was performed to repair complicated posterior urethral strictures in 3 patients. In the first procedure scar tissue was resected transurethrally to create a smooth grafting bed. In a second endourological procedure a piece of full thickness prepuce was grafted at the stricture site. An intraluminal balloon catheter was used to keep the skin graft in close contact with the resected area of the urethra. Of the patients 2 have remained free of stricture for more than 22 months and 1 has remained free of stricture for more than 12 months after endourethroplasty. The technique offers a promising alternative to open surgery in selected patients with complicated posterior urethral strictures. PMID- 3045343 TI - Post-transplant renal artery stenosis: a cause of anuria. Report of 2 cases corrected by revascularization. AB - We report 2 cases of severe hypertension and acute onset of anuria after renal transplantation in which angiography revealed renal artery stenosis. After renal artery reconstructive surgery renal function returned to normal and the hypertension improved. A high index of suspicion is needed to make the diagnosis. Only by heightened awareness of this important entity will patients with post transplantation anuria secondary to renal artery stenosis be identified. Such patients may benefit from renal artery revascularization to reverse this type of renal failure. PMID- 3045345 TI - Detection of tumor cells in bladder washings by a monoclonal antibody to human bladder tumor-associated antigen. AB - We have conducted two studies to evaluate the efficacy of using a specific monoclonal antibody (McAb) to detect exfoliated tumor cells in bladder washings. This is a preliminary step toward the development of immunological methods to improve the cytologic detection of bladder carcinoma. In this study, McAb 3G2-C6 was used. The McAb reacts to a bladder tumor-associated cell-surface antigen expressed in bladder tumors of various grades. Bladder washings from patients with and without carcinoma were stained with the McAb using two different indirect immunofluorescence methods (Methods A and B). The results of the immunological studies were compared with those obtained from the cytology laboratory and these in turn, were evaluated against the histopathological diagnosis of respective patients at the time the samples were taken. Immunofluorescence method A detected positive cells in 87% (56/64) of specimens from bladder cancer patients, including 18 of 19 from patients with grade 1 tumor. This method also had a low false-positive rate; only one of 17 specimens from patients with other urinary disorders had positively reacting cells. Immunofluorescence method B, evaluating a second group of specimens, detected positive cells in 68% (15/22) of specimens from patients with carcinoma, and in only one of 17 controls. However, it also identified positive cells in specimens from patients with chronic cystitis and urinary calculi. Overall the results of these studies indicate that the McAb method is superior to the routine cytology in detecting tumor cells in bladder washing specimens. More work must be done, however, to improve the specificity of the method before it can be used as an aid for routine tumor detection. PMID- 3045346 TI - Rabies and rabies control in striped skunks (Mephitis mephitis) in three prairie regions of western North America. AB - The number and geographic distribution of rabies cases in striped skunks (Mephitis mephitis) from Saskatchewan (n = 2,506 cases), Montana (n = 1,142), and Alberta (n = 199) since 1963 were reviewed. In Saskatchewan the number of cases increased steadily for 5 yr and then fluctuated consistently in a 4 yr cyclic pattern. Similarly an initial sweep across the province was followed by a cyclic pattern of geographic expansion (3 to 4 yr) and reduction (1 to 2 yr). No organized control efforts were conducted in Saskatchewan. Similar cyclic pattern were not seen in data from Montana or Alberta. In the latter areas, the number and distribution of rabies cases in skunks appeared to reflect efforts to reduce the population of skunks. An integrated program of skunk removal using poison and live-traps in association with research and public education successfully contributed to limiting the spread and establishment of rabies in striped skunks within prairie habitats. Rabies did not persist in skunks in other habitats. PMID- 3045348 TI - Klebsiella pneumoniae as a cause of pneumonia and septicemia in a civet kitten (Civettictis civetta) in the Jos Zoo, Nigeria. AB - A fatal case of acute pneumonia and septicemia that occurred in a captive civet kitten (Civettictis civetta) in the Jos Zoo, Nigeria is reported. Diagnosis was based on clinical signs, necropsy findings, and the isolation of Klebsiella pneumoniae from the lung, intestine, liver and heart blood of the animal. This is the first report of clinical K. pneumoniae infection in a wild or captive animal in Nigeria. PMID- 3045349 TI - Are veterans really different? The feasibility of smoke-free VA hospitals. PMID- 3045347 TI - Immunization of arctic foxes (Alopex lagopus) with oral rabies vaccine. AB - Arctic foxes (Alopex lagopus) were successfully immunized against rabies using an orally-administered, liquid SAD-BHK21 live virus vaccine in a sausage bait. Immunization was determined by serologic response and by resistance to challenge with an arctic rabies virus strain. Virus was not shed in saliva following oral vaccination, indicating that arctic foxes would not infect other foxes after ingesting this vaccine. High antibody levels were present in all experimental foxes 2 wk following initial vaccination. A booster vaccination at 56 wk induced a significant serologic response within 1 wk, suggesting an anamnestic response but titers began to decline within 8 wk in most foxes. Foxes were observed for 16 mo following the challenge and exhibited no symptoms of rabies. The SAD-BHK21 rabies vaccine in a sausage bait system has a strong potential for vaccinating wild populations of arctic fox. PMID- 3045350 TI - Medicare as secondary payer. PMID- 3045352 TI - Beginning a career in pediatric hematology: 1926. PMID- 3045353 TI - Intangibles in medicine: an attempt at a balancing perspective. PMID- 3045351 TI - Minimal contact treatment for smoking cessation. A placebo controlled trial of nicotine polacrilex and self-directed relapse prevention: initial results of the Stanford Stop Smoking Project. AB - To determine the effectiveness of nicotine polacrilex combined with self administered relapse prevention materials in maintaining smoking cessation, we conducted a randomized, double-blind, placebo controlled trial. Volunteers aged 18 to 65 years responding to media announcements were required to quit smoking for 48 hours without assistance. Of 1844 potential participants, 136 were medically excluded, 535 declined to make a quit attempt, and 573 were unable to quit, leaving 600 participants (35%) who were randomized. Eight self-help relapse prevention modules were mailed weekly. Gum was used either ad lib for smoking urges or on a fixed, hourly schedule (12 pieces per day). Only 15% of the subjects in each gum group stopped using the gum altogether because of side effects, but only 20% of the ad lib groups and 40% of the fixed-dosage group used at least eight pieces of gum per day during the first week. The abstinence rates (for at least seven days) at the six-month follow-up were 31% in both active gum groups and 22% in the placebo and no gum groups. Relapse rates in the two active gum groups were about half those in the placebo and no gum groups. Nicotine polacrilex may be a useful adjunct to minimal contact smoking cessation formats, which have broad appeal. Also, minimal contact relapse prevention programs may assist physicians in helping patients to maintain smoking cessation using nicotine polacrilex. PMID- 3045354 TI - Anencephalic organ donor program suspended; Loma Linda report expected to detail findings. PMID- 3045355 TI - From the Health Care Financing Administration. PMID- 3045356 TI - The quality of care. How can it be assessed? AB - Before assessment can begin we must decide how quality is to be defined and that depends on whether one assesses only the performance of practitioners or also the contributions of patients and of the health care system; on how broadly health and responsibility for health are defined; on whether the maximally effective or optimally effective care is sought; and on whether individual or social preferences define the optimum. We also need detailed information about the causal linkages among the structural attributes of the settings in which care occurs, the processes of care, and the outcomes of care. Specifying the components or outcomes of care to be sampled, formulating the appropriate criteria and standards, and obtaining the necessary information are the steps that follow. Though we know much about assessing quality, much remains to be known. PMID- 3045357 TI - Microcirculation and traditional Chinese medicine. PMID- 3045358 TI - A piece of my mind. Hunger. PMID- 3045359 TI - Renal vein plasma renin activity in patients with unilateral renovascular hypertension. AB - Plasma renin activity in the renal veins (V) or infrarenal inferior vena cavae (IVC) of 20 patients with unilateral renovascular hypertension (RVH) was measured to determine how renal vein renin ratio (RVRR) compares with renin index (V IVC/IVC) as a predictor of curability of RVH. Although there was no significant difference between them in predicting curability, 3 out of 4 patients with hypersecretion of renin (V-IVC/IVC greater than or equal to 0.48) in the stenosed side with contralateral suppression (V-IVC/IVC less than or equal to 0) on the normal side were cured. In addition, 7 out of 11 patients with contralateral suppression irrespective of values of renin index in the stenosed side were also cured. On the other hand, only one out of 6 patients who had neither hypersecretion nor contralateral suppression was cured. These results reconfirm that significant renin secretion with or without contralateral suppression, or only contralateral suppression of renin is highly suggestive of curable RVH. PMID- 3045360 TI - Quadricuspid aortic valve perforation resulting from bacterial endocarditis--2-D echo- and angiographic diagnosis and its surgical treatment. AB - A 75-year-old man with quadricuspid aortic valve regurgitation affected by bacterial endocarditis is reported. The aortic valve consisted of 4 equal-sized cusps (type a) and a supernumerary cusp located between the right and noncoronary cusps. A right coronary ostium was close to the accessory commissure, but there was no displacement. A few small fenestrations were found at the 4 commissures and a large perforation resulting from endocarditis was observed in the noncoronary cusp. 2-D echocardiogram and angiogram suggested these findings, and they were confirmed at surgery. Successful aortic valve replacement was achieved. PMID- 3045361 TI - [Changes in aldosterone and PRA due to the administration of potassium canrenoate in laparotomized patients with liver cirrhosis]. PMID- 3045362 TI - [Anesthetic management for subtotal pancreatectomy in a neonate with nesidioblastosis]. PMID- 3045363 TI - [Cytokines and receptors--their functions, structures and cloning of code genes. Interleukin-3]. PMID- 3045364 TI - [Problems in the clinical application of human-recombinant cytokines. Production of human recombinant cytokines and application to genetic recombination]. PMID- 3045365 TI - [Expectations in the clinical application of human-recombinant cytokines. 2) Clinical trials of IL-2]. PMID- 3045366 TI - [Oncogenes of colonic neoplasms]. PMID- 3045367 TI - [Application of mechanochemical systems to medicine. Development of artificial muscle models using ultra water-absorptive gels]. PMID- 3045368 TI - [Scanning tunneling microscope and the possibility of its application to medical use]. PMID- 3045369 TI - [Gastrointestinal motility of the large intestine and its regulation]. PMID- 3045370 TI - [Gelatin particle agglutination test for serum antibodies to HIV]. PMID- 3045371 TI - [Cytokines--DNA to immunologic diseases]. PMID- 3045372 TI - The validity of magnetic resonance imaging (MRI) in the staging of bladder cancer: comparison with computed tomography (CT) and transurethral ultrasonography (US). AB - The magnetic resonance imaging (MRI) of bladder cancer was accomplished in 26 patients who had histologically proven transitional cell carcinoma through transurethral resection. All 26 patients were simultaneously studied by computed tomography (CT) and 21 were also studied by transurethral ultrasonography (US) before surgery or preoperative irradiation. The evaluations of the MRI and CT were carried out by the same radiologist and urologist postoperatively; the evaluation of the US was carried out by several urologists preoperatively, and their results compared with the postoperative pathological findings. The staging accuracy for pT2 tumors and above for each imaging modality was as follows. MRI: sensitivity 83.3%, specificity 71.4%, accuracy 76.9%; CT: sensitivity 58.3%, specificity 42.8%, accuracy 50%; US: sensitivity 100%, specificity 70%, accuracy 85.7%. The results of the staging with regard to extravesical extensions were as follows. MRI: sensitivity 60%, specificity 85.7%, accuracy 80.8%; CT: sensitivity 60%, specificity 81%, accuracy 76.9%. On the other hand, the sensitivity of US was 25%, specificity 94.1% and accuracy was 81%. The higher the pathological staging, the more the diagnostic accuracy was likely to be increased with MRI and CT, but diminished for US. There were three cases of pT1 tumors being over diagnosed by US, but two of them were accurately staged by MRI. The other 11 cases, diagnosed as T2 or above by US, were only minimally overstaged or understaged by MRI. MRI seemed able to overcome the disadvantage of US which tended to over-diagnose pT1 tumors and which was not suitable for use in diagnosing infiltrative tumors correctly. CT seemed useless for local staging. PMID- 3045373 TI - A new hemostatic procedure for percutaneous transhepatic portal vein catheterization. AB - To avoid major complications, such as intraperitoneal bleeding after percutaneous transhepatic catheterization, a new technique has been developed. In this procedure, a stick-like gelfoam sponge compressed manually and loaded in a cartridge, is inserted through an outer sheath catheter after the removal of the catheter, and the puncture hole is plugged. No bleeding has been experienced in 56 patients with various hepatic diseases since using this method. PMID- 3045374 TI - Two cases of lipid-secreting carcinoma of the breast: case reports with an electron microscopic study. AB - Two very rare cases of lipid-secreting carcinoma of the breast are reported, together with a review of the literature. Both patients were Japanese women aged 70 and 81 years, respectively. In each, a mass had been found in the breast, and radical mastectomies were performed based on a diagnosis of breast cancer at an early clinical stage. Both resected tumors were found to be solid and localized. In them, both estrogen and progesterone receptors were negative. Microscopically, hematoxylin-eosin staining showed the tumor cells to have foamy and vacuolated eosinophilic cytoplasm. Lipid staining showed the vesicles in the cytoplasm of the tumor cells to be colored a flaming red and consequently recognized as lipid droplets. An electron-microscopic study showed that the tumor cells contained various sizes of lipid droplets in the cytoplasm with no degenerative changes, and suggested that these lipid droplets were produced and stored in the tumor cells. It would appear, from the literature that lipid-secreting carcinomas have aggressive clinical courses, but neither vascular invasion nor lymph node metastasis was seen in either of our two cases. PMID- 3045375 TI - [A case of focal dermal hypoplasia--a report of partial expressed form with a review of literatures]. PMID- 3045376 TI - [Double filtration plasmapheresis is effective in the treatment of 3 cases of autoimmune bullous disease]. PMID- 3045377 TI - Malaria in Korea. AB - Malaria was steadily decreasing in Korea except in certain counties of the mountainous and hilly areas, in the 1960s. Judging from the present epidemiological, social and economic conditions, it can be said with confidence that malaria with "unstable" characteristic in the Republic of Korea has already been disappeared. No doubt, the causes of the disappearance of malaria are complex. Certainly improved living conditions and life style; better medical and educational facilities in the wake of a rapid economic development could all have some role. On the other hand, the disappearance of malaria without large scale control operations could be ascribed to the two main factors: one is malaria case detection and simultaneous drug therapy available through the nation-wide passive case detection network during the 1960s and the other is rapidly improved farming practices begun in 1970s, which resulted in the use of a huge quantity of pesticides and other chemicals for agriculture, which, in turn, might affect local anopheline vectors which were originally not effective ones any way. PMID- 3045378 TI - [Reports on genetic markers and new polymorphism of human body fluid and its medico-legal application]. PMID- 3045379 TI - [A method for placing a temporary atrial pacing electrode in open heart surgery]. PMID- 3045380 TI - [A case of right atrial lipoma]. PMID- 3045382 TI - Cocaine-induced chest pain and myocardial ischemia. PMID- 3045383 TI - Mechanism and pattern of injury associated with use of seat belts. PMID- 3045381 TI - Interventions for flail chest questioned. PMID- 3045384 TI - Reimbursement realities and the triage nurse's role. PMID- 3045385 TI - Emergency nurses' knowledge of legal liability. PMID- 3045386 TI - Suicide by vehicular crash: recognition and early intervention. PMID- 3045388 TI - The MMWR file. PMID- 3045387 TI - Malpractice insurance--law and the emergency nurse. PMID- 3045389 TI - Minneapolis prehospital do-not-resuscitate form provides model for others. PMID- 3045390 TI - A twenty-month-old infant with a one-day inability to use one arm. PMID- 3045391 TI - Like family. PMID- 3045392 TI - Emergency Nurses Association position statements. PMID- 3045393 TI - New York State emergency medical service (EMS) setting trends. PMID- 3045394 TI - [Natural antithrombotic mechanisms. I]. PMID- 3045395 TI - [Natural antithrombotic mechanisms. II. The fibrinolytic system]. PMID- 3045396 TI - [Electroablation, a new "semi-invasive" method of treating cardiac rhythm disorders]. PMID- 3045397 TI - [Prevention of thrombophlebitis of deep veins of the legs]. PMID- 3045398 TI - [Effect of aminophylline on the heart conduction system in patients with paroxysmal nodal tachycardia]. PMID- 3045399 TI - [Evaluation of the hypotensive effect of metoprolol (Betaloc-Astra)]. PMID- 3045400 TI - [Thrombocytes during physical exertion and induced myocardial ischemia]. PMID- 3045401 TI - [Current concepts of the pathogenesis of familial hypercholesterolemia]. PMID- 3045403 TI - [Current problems of the treatment of disorders of the rheological properties of the blood in patients with ischemic heart disease]. PMID- 3045402 TI - [Morphologic manifestations of the medicinal lysis of a coronary artery thrombus]. AB - Morphologic studies of coronary arterial thrombi have shown immature fibrin fibres to prevail in those, with fibrin degradation products always present. This fact suggests that fibrinolytic components of hemostasis, limiting the rates of fibrin polymerization and maturation, may participate in thrombogenesis. At the same time, macrophages that phagocytose fibrin and blood cells (platelets and erythrocytes) become activated since the early hours of the thrombus' existence. Selective administration of fibrinolytic activators destroys the fibrin network of the thrombus, blood cells are washed away by the blood flow and the thrombus diminishes in volume. At the same time, its macrophagal response is activated, and fibrin and fibrin degradation products are eliminated from the vascular lumen. PMID- 3045404 TI - [Neurogenic regulation of the hemostatic system in ischemic heart disease]. PMID- 3045405 TI - [First aid and infusion therapy in thermal injuries in children]. PMID- 3045406 TI - [Prof. Dr. Arvo Ylppo--100 years old]. PMID- 3045407 TI - [Otto Heubner as Emeritus in Dresden-Loschwitz 1913-1926]. PMID- 3045408 TI - [Urological complications in patients following kidney transplantation]. PMID- 3045410 TI - [The potentials of a Doppler study in acute surgical diseases of the scrotal contents]. PMID- 3045409 TI - [Aseptic osteonecrosis in patients with a transplanted kidney]. PMID- 3045411 TI - [Late changes in the structure of skin autografts of the lower extremities following burns]. PMID- 3045412 TI - [The use of microsurgery in urology]. PMID- 3045413 TI - [Perioperative antibacterial prophylaxis and its use in abdominal surgery]. PMID- 3045414 TI - [Basic treatment principles in burns of the lower extremities]. PMID- 3045415 TI - [Enteral dialysis--an etiopathogenetic method for the treatment of peritonitis]. PMID- 3045416 TI - [Ultrasonographic observations of the vitreous body in perforating retinal detachment]. PMID- 3045417 TI - [From the bibliographer's files (XXXII). 150, 125 and 100 years ago]. PMID- 3045418 TI - [An analysis of research on relaxation technics reported in academic journals 1971-1987]. PMID- 3045419 TI - Junctional transfer in wounded cultures of bovine aortic endothelial cells. AB - Both in vivo and in vitro, endothelial cells form continuous monolayers displaying growth control by cell density as well as cell contact. Several functions of endothelium are dependent upon maintenance of an intact monolayer. When such a monolayer is injured, endothelial cells migrate out from the wound edges and proliferate to cover the area of denudation. The response of endothelium to wounding is therefore a matter of both growth control and cellular motility. Since intercellular gap junctions have been postulated to have a role in growth control, we set out to determine whether junctional communication was altered in the activated, migrating cells. We found extensive transfer of microinjected fluorescent dye molecules (Lucifer yellow CH) in cells at or near the wound edge at various times after wounding (1 minute to 48 hours). However, cells near a wound edge transferred dye at a frequency significantly diminished from that observed in undisturbed monolayers (81.1 +/- 1.5 (SE) versus 92.7 +/- 0.8% of adjacent cells received dye). There was a significant positive correlation between transfer frequency and distance from the wound (expressed as intervening cells) but not between transfer frequency and time after wounding. These results suggest that endothelial cells in a wounded monolayer retain junctional communication at a slightly but significantly reduced level and that increased motility and the wound healing response do not necessarily correlate with total loss of communication. PMID- 3045420 TI - Adolescent pregnancy: a team approach. PMID- 3045422 TI - Federal largesse. PMID- 3045421 TI - The families of Tennessee in the Southern Surgical Association. PMID- 3045423 TI - Effect of adjuvant chemotherapy on time to recurrence and survival of stage I uterine sarcomas. AB - We have evaluated the effect of adjuvant chemotherapy on time to recurrence and survival in two prospective trials of women with stage I uterine sarcomas. The first trial compared surgery only to surgery plus Adriamycin. The 5-year estimated survival rate was 36% for surgery alone and 63% for surgery plus Adriamycin. The 5-year recurrence free rate for surgery alone was 46% as compared to 75% for surgery plus Adriamycin. The second trial, without a concurrent control group, included patients with stage I uterine sarcoma and adjuvant cyclaphosphamide, vincristine, Adriamycin, and dacarbazine (CYVADIC) chemotherapy. The 5-year survival rate was 89% and the recurrence-free rate was 80%. In all of these trials, as well as in the report of Van Nagell et al (Cancer 57:1451-1454, 1986) of adjuvant vincristine, actinomycin-D, and cyclophosphamide (VAC) chemotherapy, there are too few patients to make any formal statistical comparison of the groups, although the surgery plus CYVADIC group appears to be the most promising. PMID- 3045424 TI - A role for topical 5-fluorouracil therapy in melanoma. AB - Topical 5-fluorouracil (Efudex) has been shown to be of great value in the treatment of skin cancers, Marjolin ulcers, and advanced squamous cell carcinoma of the mouth and esophagus. It has been advocated for use on patients with melanocytic dysplasia and lentigo maligna melanoma. This report confirms the use of topical 5-FU for lentigo maligna melanoma and melanocytic dysplasia. In melanomas with poor prognosis, preoperative treatment with topical 5-FU converts the cellular infiltrate from acute inflammatory cells to round cells which have been shown to be T-cells. We believe this preoperative stimulation of the patients' own immune reaction to the tumor may be of great significance in improving the surgical results for melanomas with a poor prognosis. PMID- 3045425 TI - Southern analysis of human head and neck cancer cells for homologous sequences using yeast gamma repair genes. AB - We employed southern analysis to examine homology between yeast genes RAD 51, RAD 52, RAD 54, and RAD 55 for possible gamma repair genes in radioresistant or repair proficient human tumor cell lines and normal placental DNA. No homology wa observed; however, other strategies including further gene restriction and transfection are underway to identify repair genes in human tumors. Understanding mechanisms of radiation repair might lead to more effective clinical radiation treatment protocols. PMID- 3045426 TI - Primary pulmonary rhabdomyosarcoma in a child, with a review of literature. AB - A cyst in the left lower lobe of the lung was removed from a 15-month-old girl and showed evidence suggesting a preexistent congenital cystic adenomatoid malformation; in addition a rhabdomyosarcoma was identified. Primitive mesenchyme and mesenchymal elements also were seen. The origin of the rhabdomyosarcoma is discussed. The findings in the present case are compared with those of all childhood pulmonary rhabdomyosarcomas reported in the literature. This report of a child with a coexistent congenital adenomatoid malformation and a rhabdomyosarcoma is the third of its kind in the literature. PMID- 3045427 TI - Prostacyclin and thromboxane formation following chronic exposure to cigarette smoke condensate administered via osmotic pumps in rats. A method for chronic administration of particulates of whole smoke. AB - Chronic cigarette smoke exposure in vivo causes decreased conversion of [14C]arachidonic acid (AA) to prostacyclin (PGI2) by isolated aortic tissue and increased conversion to thromboxane (TXA2) by isolated platelets from rats. Alterations in the PGL2/TXA2 balance may be part of the mechanism by which smoking increases the risk of cardiovascular disease. In order to ascertain whether the particulate phase of whole smoke alone could cause these changes, rats were administered smoke condensate in propylene glycol for 56 days via two Alzet (2ML4) osmotic pumps. Pumps containing vehicle, low dose (150 micrograms/hr) or high dose (300 micrograms/hr) condensate were implanted s.c. dorsal to the thoracic vertebrae in male Sprague-Dawley rats. Three-quarters of the condensate-treated rats developed fibrin cysts encapsulating the pumps. Cysts were not seen in vehicle-treated rats. Residual pump contents were weighed and analyzed by GLC to ensure condensate delivery. No significant difference in weight gain patterns between sham-operated and treatment groups were observed. Vehicle had no effect on aortic PGI2 or platelet TXA2 formation compared to sham. Low-dose condensate was without effect on PGI2/TXA2 formation. In high-dose condensate-treated rats, PGI2 and TXA2 formation were 84% and 136%, respectively, of the vehicle control (n.s.). Pump encapsulation may be a limiting factor in the administration of complex particulate suspensions. PMID- 3045428 TI - Divided right atrium (prominence of the eustachian and thebesian valves). AB - Division of the morphologically left atrium (cor triatriatum) is a recognized clinical and surgical entity. Division of the right atrium (prominence of the eustachian and thebesian valves) is recognized pathologically, but is rare. It is unusual for this entity to be diagnosed during life. Stimulated by our experience with two patients seen at operation, one with an obstructive spinnaker-like formation and the other with a partitioned right atrium in the setting of pulmonary atresia, we reviewed the specimens in the heart museum of Children's Hospital of Pittsburgh that had prominence of the eustachian and thebesian valves. We identified 14 such hearts, which could be divided into two groups. In the first group, comprising six hearts, the valves were prominent in the form of Chiari networks and were of no functional significance. The valves were more extensive in the other eight hearts and partitioned the right atrium. In two of these, the valves themselves were the impediment to flow through the right side of the heart. In the other six, there was either atresia or severe stenosis along the right-sided pathways so that, after birth, the prominent valves retained their role during fetal life; namely, to deflect inferior caval venous return across the atrial septum to the left atrium. The partitions in these latter hearts would be of functional significance only if it were necessary to perform a Fontan procedure, when they might obstruct flow through an atriopulmonary (or atrioventricular) anastomosis. PMID- 3045429 TI - The American Association for Thoracic Surgery 1988-1989. PMID- 3045430 TI - Wang Bing, annotator of 'plain questions'. PMID- 3045431 TI - Autologous bone marrow transplantation in acute lymphoblastic leukemia- preclinical immunologic studies. AB - Immunologic aspects of autologous bone marrow transplantation (ABMT), immunodiagnosis, patient monitoring, and the purging of bone marrow have been studied in individual patients. It was demonstrated that the most sensitive method for detecting lymphoid cells which show the phenotypes of ALLs of B or T lineage was double immunofluorescence staining for nuclear terminal transferase (TdT) and B or T lineage antigens. With the help of these sensitive tests in the presence of rabbit complement (C'), MAbs CD10 (RFAL3 of IgM class), CD19 (SB4 of IgM class), and their cocktail were capable of eliminating greater than 3 log blast cells of B lineage ALL in 84%, 75.5%, and 90% of cases, respectively. The same reagents lysed 26.8%, 0%, and 45% of blasts in the presence of human C'. CD7 (RFT2, IgG2) eliminated greater than 3 log T-ALL blast cells in 73% of cases. The proliferative fractions of leukemic blasts were also TdT+ and sensitive to lysis with MAb and C'. On the basis of these observations MABs were selected for purging in 36 patients undergoing ABMT in first remission (10 patients considered to be at a high risk of relapse), second and third remissions (23 and 2 patients), and without entering into remission (1 patient). The efficacy of eliminating the MAb-reactive cells from the bone marrow inoculum was also documented in five patients. By the use of sensitive immunologic assay (TdT/cytoplasmic CD3 double staining) in patients with T-ALL, no residual leukemia (less than 10(-4] could be detected at the time of transplantation. Following an observation period of 5-34 months, 24 of the 36 patients are alive and well with no procedure-related mortality. PMID- 3045432 TI - Cellular protein databases: a new approach to the study of genomic organization and function of the cell. PMID- 3045433 TI - 50th anniversary of the BLB oxygen mask. PMID- 3045434 TI - Surgical pathology of the tricuspid valve: a study of 363 cases spanning 25 years. AB - Surgical pathologic features of the tricuspid valve were reviewed in 363 patients who had undergone tricuspid valve replacement at our institution during the period 1963 through 1987. Valves were purely regurgitant in 74%, stenotic and regurgitant in 23%, and purely stenotic in 2%; two valves were neither stenotic nor regurgitant. Among 269 purely insufficient tricuspid valves, the four most common causes were postinflammatory disease (41%), congenital disorder (32%), pulmonary venous hypertension (21%), and infective endocarditis (4%). Of 92 cases of tricuspid stenosis, with or without regurgitation, postinflammatory disease was observed in 92%. Female patients accounted for 66% of the 363 cases, including 84% of those with postinflammatory disease and 64% of those with pulmonary venous hypertension. In contrast, male patients accounted for 73% of cases with endocarditis and 61% with congenital heart disease. Although postinflammatory disease accounted for 53% of the 363 cases, its relative frequency diminished from 79% during 1963 through 1967 to only 24% during 1983 through 1987. This trend may reflect the decreasing incidence of acute rheumatic fever reported in Western countries. During the same time interval, the relative frequency of congenital heart disease as a cause of tricuspid dysfunction increased from 7% to 53%, and it is currently the most common cause in our surgical population. This finding apparently reflects changes in patient referral practices and the development of new operative procedures. PMID- 3045435 TI - Controlled-release Sinemet (CR-4): a double-blind crossover study in patients with fluctuating Parkinson's disease. AB - Patients with moderate to advanced Parkinson's disease may have prominent levodopa-related motor fluctuations. In a double-blind crossover study, we compared the anti-Parkinson effects of standard Sinemet with a controlled-release formulation (Sinemet CR-4) in 23 patients with short-duration responses to standard Sinemet. With Sinemet CR-4, approximately half the patients who completed the study displayed a prolongation of their "on" response (optimal response to treatment), as assessed by monitoring individual drug-response cycles. A few patients experienced prolonged delays before the peak anti Parkinson response developed to Sinemet CR-4. End-of-dose "wearing off" was favorably affected by Sinemet CR-4, but patients still had unpredictable "off" (parkinsonian) periods. Subjective ratings of Sinemet CR-4 varied, and 39% of patients who completed the study actually preferred standard Sinemet to the new formulation. We conclude that Sinemet CR-4 may benefit some patients with Parkinson's disease with a short-duration response to standard Sinemet; however, not all patients found it preferable to the standard formulation. PMID- 3045436 TI - Persistent primary hyperparathyroidism: successful ultrasound-guided percutaneous ethanol ablation of an occult adenoma. AB - Ultrasound-guided percutaneous ethanol ablation of a small occult parathyroid adenoma was successfully performed in a patient with persistent primary hyperparathyroidism. This procedure may be an alternative to reoperation for patients in whom surgical treatment is contraindicated or who have an unacceptable risk of postoperative morbidity. PMID- 3045437 TI - Acute interstitial nephritis: immunologic and clinical aspects. AB - Acute interstitial nephritis is a common renal syndrome that may be associated with a variety of infections and drug therapies or may develop without an identified cause. Three cases are presented to illustrate the three types of acute interstitial nephritis--drug related, infection related, and idiopathic. Cell-mediated immune mechanisms seem to be more important than humorally mediated mechanisms in the pathogenesis of acute interstitial nephritis. Frequently, eosinophils are identified as a component of the interstitial cellular infiltrate, and eosinophiluria and eosinophilia have been claimed to be helpful in the diagnosis of acute interstitial nephritis, especially the drug-induced type. Neither eosinophiluria nor the presence of increased urinary levels of eosinophil major basic protein, however, is specific for the diagnosis of acute interstitial nephritis. Patients with drug-induced interstitial nephritis frequently have symptoms and signs suggestive of a hypersensitivity syndrome and rarely have more dramatic anaphylactic manifestations. Systemic glucocorticoids have been shown to be beneficial in this type of acute interstitial nephritis. PMID- 3045439 TI - The tricuspid valve. PMID- 3045440 TI - A historical perspective of statistics. PMID- 3045438 TI - Signal-averaged electrocardiography: a new noninvasive test to identify patients at risk for ventricular arrhythmias. AB - Signal-averaged electrocardiography (ECG) is a new noninvasive test for identifying patients at risk for ventricular arrhythmias. This computerized method of analyzing standard ECGs identifies particular microvolt-level signals called late potentials. Late potentials have been correlated with clinical ventricular tachycardia, are predictive of ventricular tachycardia inducibility at the time of electrophysiologic testing, and are predictive of arrhythmic events after myocardial infarction. In this review, we describe late potentials, the method of obtaining and processing the signal-averaged ECG, and clinical studies in various patient groups that have assessed the predictive value of the signal-averaged ECG for identification of patients at risk for subsequent ventricular arrhythmias. PMID- 3045441 TI - Double-blind, placebo-controlled lithium treatment in chemotherapy induced aplasia for AML: reduced antibiotic requirement. AB - A double-blind placebo-controlled study on lithium (Li) therapy after chemotherapy-induced bone marrow aplasia was undertaken in 53 patients with acute myeloblastic leukemia (AML). No difference was observed between the two groups for the duration of aplasia, the number of units of platelets or RBC transfused, the complete remission rate or the disease free survival. However, a statistically significant reduction in the number of days of antibiotic therapy required was found in the treated group (10.55 +/- 2.72 vs 12.73 +/- 3.60, P less than 0.05). PMID- 3045442 TI - Luminol-enhanced chemiluminescence of peripheral blood leukocytes as an early indicator of graft take after allogeneic bone marrow transplantation in patients with acute myelogenous leukaemia. AB - The luminol-enhanced chemiluminescence (CL) of peripheral blood leukocytes was studied daily in five patients with acute myelogenous leukaemia (AML) in first remission, who were undergoing allogeneic bone marrow transplantation (BMT). The CL was measured after stimulation of leukocytes with opsonized zymosan in highly diluted whole blood. All patients had an undetectable CL level on day +7, post BMT, simultaneously with severe pancytopenia caused by the pre-conditioning for BMT. Subsequently, CL started to rise, reaching the maximum level, twice that of healthy controls, on day +11. This preceded the rise of blood leukocytes above 1.0 x 10(9) l.-1 and that of neutrophils above 0.5 x 10(9) l.-1 by 3-14 days, but coincided with the appearance of large unstained cells (LUC; a parameter given by a Technicon H 6000 blood analyzer). One of the patients later had a transient decline of CL. This preceded the fall in white blood count and platelets by 7 days, suggesting marrow suppression. We conclude that in AML the measurement of leukocyte CL is a more sensitive test for prediction of graft take than the conventional blood counts. PMID- 3045444 TI - [B-cell lymphomas. An immunohistological study with monoclonal antibodies in 197 cases]. PMID- 3045446 TI - [Diagnosis of B-cell lymphomas]. PMID- 3045445 TI - [A double-blind controlled clinical trial of nicardipine versus placebo in acute cerebral focal ischemia]. PMID- 3045443 TI - Mechanisms of cellular resistance to cisplatin. AB - Treatment of cancer patients often fails because of the resistance in the tumor to chemotherapeutic drugs. A better understanding of mechanisms which are active in resistant cells might lead to measures to circumvent the resistance. This review deals with the mechanisms of action of cisplatin (CDDP) and the various causes for CDDP resistance in the tumor cells. Also, possibilities to circumvent CDDP resistance in vitro and in vivo are presented. PMID- 3045448 TI - [Multiple myeloma and idiopathic monoclonal gammapathy. Diagnosis, prognosis and treatment]. PMID- 3045447 TI - [Management of craniocerebral trauma in a regional hospital]. PMID- 3045449 TI - Identification of persons at risk for sudden cardiac death. AB - Before any more progress is made in reducing the incidence of sudden cardiac death, our ability to identify those at risk must be refined further. The close association with coronary artery disease necessitates that the first step must be the identification of those with underlying coronary artery disease. This is underscored by the disturbing fact that, in many, sudden death is the first sign of coronary disease. An aggressive evaluation of those with significant risk factors appears justified. The second part of the problem is the identification of those with coronary artery disease who are at especially high risk. The current diagnostic modalities available suffer from a relative lack of specificity to be applied indiscriminately in light of the expense and morbidity of effective therapies (that is, coronary artery bypass surgery, antiarrhythmic drugs, implantable defibrillators, surgical or catheter ablation). At the present time, we can identify certain subsets that warrant aggressive therapy: survivors of sudden death events or sustained ventricular tachycardia, obstructive cardiomyopathies, aortic stenosis, left main coronary artery disease, and congenital QT prolongation. Less aggressive but also less specific therapies, such as beta-blockers in myocardial infarction survivors, can be given more indiscriminately. Ultimately, of course, the greatest impact will come from prevention of coronary artery disease. PMID- 3045450 TI - Silent myocardial ischemia: an update. AB - Silent myocardial ischemia is diagnosed by several different techniques and has been documented in all the anginal syndromes. In addition to other factors, its presence may be related to increased pain threshold and increased pain tolerance. Although some patients with painless ischemia may have less extensive coronary artery disease, cumulative evidence indicates that silent myocardial ischemia does not necessarily signify a lesser degree of cardiac ischemia or a less severe coronary abnormality. As judged by ambulatory monitoring studies, it shows circadian variation; occurs more frequently than symptomatic ischemia; and appears to depend, in large part, on activation of the sympathetic nervous system. Frequent silent ischemic events during ambulatory monitoring are worrisome because they reflect the disease "activity" of single or multiple coronary atherosclerotic lesions. Thus, there may be a direct association between the severity of ischemia seen during Holter monitoring, the extent of underlying coronary artery disease or disease activity, and prognosis. When diagnosed by exercise testing, silent myocardial ischemia may be associated with significant coronary involvement. In this regard, patients with three vessel coronary disease, impaired left ventricular function, and silent ischemia during stress testing should benefit from coronary revascularization. Compared with symptomatic patients, other evidence suggests that patients with exercise-induced asymptomatic ischemia have at least the same or perhaps even a worse outlook; this may be related to the lack of symptoms that would prompt evaluation and therapy. Awareness of the possibility of silent myocardial ischemia and use of commonly available tests, both to establish its presence and severity and to guide treatment, are emerging as new clinical goals. Further data, however, are necessary to determine how vigorously this should be pursued in different patient subgroups. In association with unstable angina or post-myocardial infarction, the added risk of silent myocardial ischemia warrants a more aggressive approach. PMID- 3045452 TI - Mesenteric ischemia. AB - Superior mesenteric artery embolism or thrombosis and nonocclusive ischemia are the most frequent causes of mesenteric ischemia. Symptoms out of proportion to the physical findings, leucocytosis, and metabolic acidosis suggest the diagnosis. A high index of suspicion, aggressive resuscitation and correction of metabolic derangements, early angiography, and operative intervention are necessary if the current high mortality rates are to be reduced. PMID- 3045453 TI - Primary hyperaldosteronism. AB - Primary hyperaldosteronism is a challenging diagnosis because of its low incidence and variable pathophysiology. Serum potassium, properly done, is the routine screening test, but is not without its limitations. Confirmation of the diagnosis requires demonstration of abnormally high and nonsuppressible values for aldosterone in plasma and urine and low plasma renin activity. Sophisticated biochemical profiling and localization procedures often are required to identify those subtypes that will benefit from surgical management, including aldosterone producing adenomas, primary adrenal hyperplasia, unilateral hyperplasia, and aldosterone-producing renin responsive adenomas. Glucocorticoid-suppressible hyperaldosteronism and isolated aldosterone-producing adrenal carcinoma are rare additional subtypes to be identified. Differentiation among these subtypes is a developing process that can be expected to continue to improve with new techniques and new understanding of underlying pathophysiology. PMID- 3045451 TI - Diagnosis of pulmonary infections in immunocompromised patients. AB - In an immunocompromised patient with fever and pulmonary infiltrates, it frequently is difficult to decide which invasive procedure, if any, to use to obtain a definitive diagnosis. Because most lung infiltrates in immunosuppressed patients are caused by bacteria and sputum usually is readily available for examination, empiric therapy with potent, safe, broad spectrum, antibacterial drugs often is successful. Invasive procedures that prove a diagnosis may result in substantive changes in therapy in perhaps as few as 10 to 20 per cent of patients, and the procedure itself may harm the patient. In a unique study in which patients with acute pneumonitis without neutropenia were randomized to either empiric antibiotic treatment or treatment based on results of open lung biopsy, patients with open lung biopsy had a worse outcome, possibly related to morbidity of open lung biopsy. Furthermore, no diagnoses were provided by open lung biopsy that were not treated by the empiric regimen. A missed treatable disease may be tragic, however. A thoughtful clinician must evaluate each patient with careful consideration of the history in light of the underlying disease and its treatment, rapidity of clinical course, physical examination, and laboratory data, particularly the chest radiograph, sputum examination, and bleeding parameters. Fiberoptic bronchoscopy with washings and brushings is very safe; the addition of transbronchial biopsy adds diagnostic power at the price of some complications. Bronchoalveolar lavage is a very promising technique that probably will find widespread use. However, none of the foregoing techniques is completely sensitive. When no diagnosis is established and bronchoscopy studies are negative, open lung biopsy must be considered, especially when the chest radiograph or computed tomography scan suggests focal disease or lymphadenopathy. Needle aspiration can be used, particularly if local experience is favorable and lung disease is peripheral. When evaluating a procedure, local experience must be considered rather than reliance on published diagnostic yields and complication rates. New diagnostic and therapeutic developments may change decision analysis in the near future. At present, cultures for viruses and fungi and serologic techniques have little application at most medical centers, and decisions on data from invasive procedures pivot on interpretation of histology and smears. Development of assays for antigen (for example, Aspergillus) and rapid culture techniques (for example, cytomegalovirus and the shell vial method), coupled with new, effective antimicrobials, may demand maximum effort for a definitive diagnosis in every patient. PMID- 3045454 TI - Solitary thyroid nodule: diagnosis and management. AB - Thyroid nodules are common. Most are benign lesions since clinically important thyroid carcinoma is a relatively rare disease. The most sensitive and specific test for the diagnosis of thyroid cancer is fine-needle aspiration biopsy, but its diagnostic accuracy depends upon whether or not one excises all suspicious nodules, thus including them as correctly diagnosed. Nevertheless, fine-needle aspiration biopsy is the most sensitive, specific, and cost-effective test for thyroid cancer. Therapy depends upon the cause of the thyroid nodule. PMID- 3045456 TI - Approach to the patient with diffuse lung disease. AB - When evaluating diffuse lung infiltrates, the clinician should place special emphasis on the acuity of symptoms, nonpulmonary complaints and findings, environmental exposures, and risk factors for immunosuppressive diseases. Certain radiographic features, such as the distribution of opacities, hilar adenopathy, Kerley-B lines or pneumothorax, or pulmonary function tests demonstrating air flow limitation also narrow the differential diagnosis. One can direct the subsequent workup based on the narrowed differential diagnosis, the pace of disease, the activity of the ongoing inflammatory-immune process, and the age, overall medical condition, and wishes of the patient. Unless a specific diagnosis (for example, hypersensitivity pneumonitis, the treatment of which is withdrawal of the offending agent) can be made, therapy of noninfectious diffuse lung disease is quite unsatisfactory. Immunosuppressive therapy is indicated to arrest the active inflammatory process with the hope that objective signs of improvement will occur after a 3- to 12-month course. Important areas of basic research in pulmonary fibrosis include cell-cell and cell-matrix interactions in the lung interstitium and delineation of fibroblast biology and cytokine-mediated lung connective tissue pathology. More successful therapies will probably evolve from better understanding of the molecular and cellular biology of the lung fibrogenic process. PMID- 3045455 TI - Acute abdominal disease in the aged. AB - The elderly patient with acute abdominal disease may present with a classical clinical picture. However, the presentation often is atypical and perplexes the physician. The factors involved include altered anatomical features, fear of being placed in an institution, difficulty in communicating with the physician and family members, diminished response to infection, and multiple coexisting diseases. Awareness of the atypical clinical presentations and the judicious use of special investigations will enable the clinician to make earlier and more accurate diagnoses and, thus, reduce morbidity and mortality. PMID- 3045457 TI - Fever of unknown origin. AB - The evaluation of an FUO is a significant test of all a physician's clinical skills. The ultimate goal of the physician is to reach a diagnosis and to cure the patient in the best possible situation. Despite such pressure both externally and self-imposed, a physician needs to meticulously follow the patient and logically pursue the available diagnostic tests. To "shotgun" the process, except in the most urgent situation, is to ultimately create more frustration, confusion, and despair among the physician and his patient. PMID- 3045458 TI - Perinephric abscess: the missed diagnosis. AB - Perinephric abscess is a life-threatening but treatable process. Most infections of the perinephric space occur as a result of extension of an ascending urinary tract infection, commonly in association with nephrolithiasis or urinary tract obstruction. A large portion of the mortality is the result of failure to diagnose this entity in a timely fashion. This failure may be because of the frequently obscure or nonspecific nature of the clinical presentation. Blood cultures as well as urine cultures may fail to identify correctly the bacterial pathogens responsible for the abscess. Perinephric abscess should be considered in the differential diagnosis of any patient presenting with a urinary tract infection that fails to respond promptly to antibiotic therapy, particularly in those known to have anatomical abnormalities of the urinary tract or diabetes mellitus. Consideration of this diagnosis should enter into the differential diagnosis of fever with abdominal pain or flank pain. Early recognition of perinephric abscess and prompt drainage, either percutaneously or surgically, in combination with appropriate antibiotic coverage, should reduce dramatically the morbidity and mortality from this infection. PMID- 3045459 TI - Catecholamine-induced cardiovascular disease in the spontaneously hypertensive and atherosclerotic turkey. AB - The interest evoked by the Broad Breasted White Turkey (BBWT) as an animal model for studying the cardiovascular damages produced by hypertension and catecholamines is mainly due to the fact that hypertension is spontaneous and tissue and circulating catecholamines, especially norepinephrine, are extremely high. In this paper we focused our attention on three characteristic pathophysiological features displayed by these animals which are strictly related, as well as in humans, to the elevated blood pressure values and to catecholamine action. We also described the possibility of modifying the development of some of these lesions with pharmacological interventions liable to antagonize the peripheral effects of norepinephrine and epinephrine. The dissecting aneurysm of the aorta accounts for 5-10% of sudden deaths in this animal strain. It can be prevented by lowering blood pressure, especially with beta-blockers, and facilitated by MAO-inhibitors. The degree of cardiac hypertrophy is remarkably high and unexpectedly characterized by the synthesis of a "fast" V1-like isomyosin with high Ca++ activated ATPase activity, oxygen consumption and speed of muscle shortening. Neither the reduction of the degree of cardiac hypertrophy, nor treatment with labetalol alone were able to modify this peculiar pattern. In spite of having very high levels of high-density lipoproteins, which are known to be protective against atherosclerosis, this animal develops a severe atheromatous disease especially in the abdominal aorta, where the cellular growth has also been proven to be in vitro more pronounced than in the thoracic tract. Treatment with beta-blockers reduced the severity and extent of the lesion even in absence of a significant reduction in blood pressure. PMID- 3045460 TI - [Rapid diagnosis of group A streptococci in the throat cannot yet substitute for conventional culture technics]. PMID- 3045461 TI - [Rapid diagnosis of urinary tract infections--evaluation and recommendations]. PMID- 3045462 TI - [Rapid diagnosis of Chlamydia--new methods evaluated]. PMID- 3045463 TI - [Ecology of tick-borne infections in Fennoscandia--a review]. PMID- 3045464 TI - [Non-homogeneous hepatic fatty infiltration--a diagnostic problem]. PMID- 3045465 TI - [A physician who analyzed 60,000 medical terms promises to give more exact advice for a revised lexicon]. PMID- 3045466 TI - [A swinging chair and rolling device--treatment experiment in psychiatry]. PMID- 3045467 TI - [The collagen molecule. 2]. PMID- 3045468 TI - Evaluation of the anaphylactoid activity of a new LHRH antagonist. AB - ORF 23541 [N-Ac-D-Nal(2)1,D-pCl-Phe2,D-Pal(3)3,Ser4,Nic-Lys5,D-Nic-Lys 6, Leu7, I Lys8, Pro9,D-Ala10NH2; "Nal-Lys antagonist"] was identified as a potent LHRH antagonist without significant anaphylactoid activity. It blocked ovulation in proestrus rats when administered subcutaneously with an ED50 of 5.8 micrograms/kg. Much higher doses of ORF 23541 than of other antagonists were required to induce a cutaneous anaphylactoid-like reaction. Intradermal administration of ORF 23541 caused an 8.75 x 8.75 mm wheal response with estimated doses of 10.9 and 13.7 micrograms in rats and guinea pigs, respectively. These doses were at least 10 times greater than that required of other LHRH antagonists for the same response. ORF 23541 also did not alter pulmonary function in guinea pigs or dogs when administered intravenously at doses up to 10 mg. These results indicate that ORF 23541 represents a new generation of LHRH antagonists with an improved safety margin. PMID- 3045470 TI - [Effectiveness of different dose fractionation regimens in distant and intracavitary gamma therapy of cervical cancer]. AB - The paper is devoted to a cooperative study of comparative assessment of the efficacy of 3 dose fractionation regimens in intracavitary gamma-therapy used in combined radiation therapy of cervical cancer (5 Gy, 10 fractions, 2-3 times a week; 7 Gy, 7 fractions, once in 5 days; 10 Gy, 4 fractions, once a week). Over the period of 1979-1983 1193 patients were treated by uniform methods of intracavitary and distant irradiation. The groups were identical with relation to the number of patients, stage of disease, histological characterization and growth type. By the end of therapy the cure rate among 1183 patients was 90.3%. In 2-3 months the cure rate in the same group was 97.5%. Delayed regression was most noticeable in stages II and III. The 5-year survival rates in 3 groups were: for stage I-95.3 +/- 2.6%; for stage II-83.1 +/- 2.5%; for stage III-73.3 +/- 4.7%. In stage I and II tumors the 5-year survival rates in different regimens were similar whereas in stage III better results were obtained with a fractional dose of 5 Gy. The frequency and degree of severity of late radiation complications of the rectum, bladder, vaginal mucous membrane grew with an increase in a fractional dose. PMID- 3045469 TI - Comparison of insulin and glucagon pulsatile secretion between the rat and dog pancreas in-vitro. AB - Sustained pulses of insulin and glucagon were obtained from the isolated perfused in vitro rat pancreas. The respective periodicity of hormone release (peak to peak interval) was calculated by the Pulsar computer algorithm as insulin 5.8 +/- 0.3 min and glucagon 6.5 +/- 0.25 min. Because pulsatile insulin secretion is absent in type II diabetics, pulsatile islet hormone secretion could theoretically be regulated directly by intra-islet hormone interactions or indirectly by hormone sensitive nerve feedback, possibly from a venous hormone sensitive receptor system within the pancreas. To test the possible contributions of these systems in pulse regulation, the direction of perfusion was reversed in both rat and dog pancreata to prevent hormone contact with putative venous hormone receptors. The periodicity of hormone secretion was unchanged by reversed perfusion in both species. As vascular perfusion of islet cells is normally B to A to D, these results suggest that neither intra-islet hormone interactions nor intra-pancreatic insulin or glucagon sensitive nerve feedback systems are responsible, on an acute basis, for the regulation of pulsatile insular secretion from the normal pancreas. Insulin regulates net glucagon secretion but does not acutely influence glucagon pulses. The presence of pulses during retrograde perfusion may be the result of the entrainment of the pacemaker-islet system. These observations are consistent with the presence of an independent pacemaker and neural coordinating system within the dog and rat pancreas which may influence both the A- and B-cell. PMID- 3045471 TI - [Radionuclide and ultrasound studies in nodular (colloid) goiter]. AB - The informative value of scintigraphy and ultrasonic investigation (USI) was analyzed in 110 patients with nodular colloid goiter. The patients were operated upon, and diagnosis was made on the basis of histological findings. Scintigraphy was more preferable for the assessment of atypical position of the thyroid and function of its different parts but USI was better for the detection of its nodular, diffuse and other structural changes. Both methods contained no specific information pertaining to histology. PMID- 3045472 TI - [Radionuclide and ultrasound studies in liver cirrhosis with portal hypertension]. AB - Combined radionuclide and ultrasonic investigations (USI) were performed in 95 patients with liver cirrhosis complicated by portal hypertension. Liver and splenic shape structure and the presence of fluid in the abdominal cavity were assessed in USI. Radionuclide methods of investigation were used for determination of the hepatic blood flow, assessment of the shape, size and structure of RP distribution in the liver and spleen, and for calculation of the hepatosplenic index. The most significant signs of differentiation of stages of portal hypertension were the presence and amount of fluid in the abdominal cavity and a hepatic blood flow value reflecting the gravity of protal hypertension. Combined radionuclide and ultrasonic investigations permitted a differentiated approach to staging of portal hypertension and assessment of liver and splenic morphofunctional state that could play an important role in the choice of tactics of surgery of liver cirrhosis. PMID- 3045473 TI - [Articles from the International Journal of Radiation Oncology, Biology and Physics]. PMID- 3045474 TI - [Radiotherapy in combined and multimodal treatment of endometrial cancer]. PMID- 3045475 TI - [Soviet hospital in Algeria]. PMID- 3045476 TI - [History of the creation and development of social care for the population of Russia. The Ekaterininskaia Asylum]. PMID- 3045477 TI - Idiopathic ventricular tachycardia. A review. AB - Idiopathic or unexplained VT occurs in a small but important subset of patients without clinically evident heart disease. The majority of these patients appear to have a structurally normal heart. The cause of the arrhythmias in these individuals is unclear and may never be recognized. Other patients with this condition may have minor abnormalities not sufficient to impair overall cardiac function. The significance of these abnormalities to the genesis of the arrhythmia is uncertain. Whether patients with minor abnormalities are more likely to harbor covert heart disease such as myocarditis or a focal defect is not known, nor is it resolved whether such patients warrant a more aggressive search for a structural cause. The question that remains in any patient not subjected to surgical or pathological exploration is whether undetermined heart disease is responsible for the arrhythmia. Continued correlation between functional (electrophysiological) and structural (pathological) data will provide meaningful information concerning the pathophysiology (substrate) of these arrhythmias. Because of the preservation of normal cardiac function, these arrhythmias are generally well-tolerated. Although the condition is usually associated with a favorable prognosis, the occasional deaths reported in patients with apparently idiopathic ventricular arrhythmias may not permit calling this condition benign. It would be important to know the extent to which unrecognized abnormalities play a role in the genesis of these tachycardias, and whether such patients are more predisposed to fatal arrhythmias or have a different natural history. If cases involving undetermined or covert heart disease were excluded from consideration, then a relatively homogeneous disease-free group may be identified with a truly benign condition and a uniformly favorable prognosis. In these cases, a primary electrical abnormality may prove to be the basis for the arrhythmia. These issues remain to be elucidated in future studies. PMID- 3045478 TI - Mediastinal fibrosis complicating histoplasmosis. AB - Mediastinal fibrosis, the most serious late complication of remote infection by Histoplasma capsulatum, is a thick, dense fibrotic capsule which surrounds a small mediastinal focus of old caseous adenitis. The fibrotic process may accrue over prolonged periods and extend within the lumina of critical mediastinal structures to produce complete occlusion. We summarized clinical and radiographic data for 71 patients with mediastinal fibrosis; the criteria for inclusion were the clinical demonstration of occlusion of major central airways (trachea or mainstem bronchus) or major vessels (pulmonary arteries or veins) and the absence of other disease processes which might cause such obstruction. We selected 65 patients who met these criteria from the medical literature of the last 40 years and report 6 new cases from our experience. The majority of patients were diagnosed between ages 20 and 40. The most common symptoms included hemoptysis, dyspnea, and cough. An accentuated pulmonic component of the second heart sound, wheezing, and localized murmur were among the physical findings reported. Radiographic abnormalities consisted of mass lesions and atelectasis or infiltrates, but were often nonspecific. Chest radiography was deceptively normal in some patients, even in the presence of major central airway or vascular occlusion, especially when the focus was subcarinal. Computed tomography has particular promise to depict the mediastinal abnormalities in this process. Surgery had minimal therapeutic benefit. Because of incomplete followup, the mortality of 30% in this series surely does not represent the true overall mortality of mediastinal fibrosis. PMID- 3045479 TI - Genetic variation of glucose-6-phosphate dehydrogenase: a catalog and future prospects. PMID- 3045480 TI - Sex differences in the micronucleus test: true or false? PMID- 3045481 TI - Mouse lymphoma L5178Y thymidine kinase locus assay of 50 compounds. AB - Mutagenicity results are presented for 50 compounds tested in the mouse lymphoma TK+/(-)----TK-/- forward mutation assay. Test compounds were mostly from chemical classes not previously tested, to provide new information on the sensitivity of the assay; chemicals of low toxicity or thought to be non-carcinogenic and metabolic inhibitors, to indicate whether and under what conditions the assay can generate so-called false positive results. Twelve compounds that have been tested previously were included in this study to provide an indication of the reproducibility of the assay. Concordant results were obtained for nine of these, while disagreeing, positive results were seen with aniline, fluorene and pyrene. The following compounds belonging to the noncarcinogen category were positive at concentrations in the range 0.02-1 mol/l: dimethyl sulphoxide, EDTA, glucose, polyethyleneglycol, sodium chloride, sodium nitrite and urea. Measurements of the osmotic pressure indicated a lack of a simple relationship to mutagenic effects for these compounds. While the potent mutagenic/carcinogenic compounds tested gave greater than 4-fold increases in the mutation frequency, weak carcinogens or compounds not known to be carcinogenic that were positive in the assay gave increases of between 2- and 4-fold. Exceptions were aldehyde derivatives and chemicals that can lead to oxidative stress, which were detected with exaggerated sensitivity by the assay. Among the metabolic inhibitors tested, positive results were obtained with actinomycin D, cycloheximide, diethyl maleate, hydroxyurea and ouabain. Negative results were found with antimycin A. On the basis of the present results and previously published data it is concluded that a maximum limit for the test compound concentration can be set at 20 mmol/l and that testing to 20% total growth is adequate, with certain stipulations, to detect the mutagenic activity of test compounds. A similar analysis of the available test data shows that less than 4-fold increases in the mutation frequency have a lower predictivity for carcinogenicity. PMID- 3045482 TI - Acetaldehyde-induced aneuploidy in cultured Chinese hamster cells. AB - In this paper we report the cytogenetic activity of ethanol (EA) and its main metabolic derivative, acetaldehyde (AA) in cultured Chinese hamster cells. AA induced an increase of aneuploidy and chromosome breaks and exchanges. EA only induced an increase of achromatic lesions (gaps). However, following treatment with AA, the most notable effect was an increase of hypodiploid cells and a parallel increase of multinucleated interphase cells, the frequency of hyperdiploid cells being considerably lower. A strong correlation (r = 0.93, P less than 0.01) has been found on comparing the frequency of hypodiploid cells to the frequency of hyperdiploid plus multinucleated interphase cells. We conclude that there exists a higher chance of hypodiploid cells reaching the second mitosis after treatment and that the main effect of AA is the induction of hypodiploidy rather than hyperdiploidy and chromosome aberrations. PMID- 3045483 TI - High-dose-level effects in mutagenicity assays utilizing mammalian cells in culture. AB - We report here results obtained using sodium chloride and potassium chloride as model compounds to investigate the effects of high dose levels and osmolarity mediated artefacts in mutagenicity assays using cultured mammalian cells. Three assay systems were used with different genetic end-points: (i) mutation to 6 thioguanine resistance in Chinese hamster V79 cells; (ii) induction of chromosome aberrations in Chinese hamster ovary cells; and (iii) induction of unscheduled DNA synthesis in HeLa S3 cells. In V79 cells we observed sporadic increases in mutation frequency after treatment with sodium chloride. Potassium chloride increased the mutation frequency in a narrow interval of dose levels. Chromosome aberrations were induced by both compounds at the highest concentrations tested, confirming previously published data; the effects of potassium chloride were more marked. The chromosome aberrations observed included both deletions and exchange figures. Neither compound induced UDS in our experiments. We performed the Ames test as a check for mutagenic contaminants and both compounds gave entirely negative results. Although the osmolarities of sodium chloride and potassium chloride solutions were similar, the effects of potassium chloride were always greater at equivalent concentrations; the nature of the solute, and not only the observed osmolarity, appears to influence the results obtained. Similarly, we have observed that dimethylsulphoxide causes a marked increase in the osmolarity of the culture medium without any obvious induction of chromosome aberrations. Although the presence of S9 had little effect on the osmolarity of the medium, differences in response were observed after treatments with or without S9. For the two model compounds it appears that an osmotic mechanism alone does not provide a sufficient explanation of the findings. PMID- 3045484 TI - The genetic toxicology of some hydrocarbon and oxygenated solvents. AB - Two hydrocarbon solvents (heptane and Special Boiling Point Spirit 100/140) and eight oxygenated solvents [methyl ethyl ketone, methyl isobutyl ketone, diacetone alcohol, di-isobutyl ketone, isopropyl ether, hexylene glycol, secondary butyl alcohol and ME 6K (pentoxone)] have been tested for genotoxic activity. The solvents were tested in bacterial mutation assays, a yeast assay for mitotic gene conversion and in cultured mammalian cells (either rat liver or Chinese hamster ovary) for structural chromosome damage. All of the solvents gave a negative response in the bacterial mutation assays and the yeast mitotic gene conversion assay. In the rat liver chromosome assay, diacetone alcohol evoked a weak positive response, the remaining solvents gave a negative response. PMID- 3045485 TI - Microsomal activation of 1- and 3-nitrobenzo[a]pyrene to mutagens in Chinese hamster ovary cells. AB - 1- and 3-nitrobenzo[a]pyrene (1- and 3-nitro-BaP) are environmental pollutants and are S9-mediated mutagens in the Chinese hamster ovary (CHO) cell/hypoxanthine guanine phosphoribosyl transferase assay. In this study, we examined the pathways leading to the mutagenic activation of these compounds in CHO cells. The microsomal metabolites of 1- and 3-nitro-BaP, the 1- and 3-nitro-BaP trans-7,8 dihydroxy-7,8-dihydrodiols (trans-7,8-dihydrodiols) and the 1- and 3-nitro-BaP trans-9,10-dihydrodiols, were isolated and tested for mutagenicity. At the concentrations assayed, both trans-9,10-dihydrodiols were non-mutagenic with and without S9 activation. In contrast, the trans-7,8-dihydrodiols of 1- and 3-nitro BaP were direct-acting mutagens and these responses were similar in magnitude to the S9-mediated mutagenicities of the parent nitro-BaPs. S9 increased the mutagenic responses of the trans-7,8-dihydrodiols approximately 20-fold. Inhibition of epoxide hydrolase decreased the S9-mediated mutagenicity of 1-nitro BaP by half, but doubled the S9-mediated mutagenicity of 3-nitro-BaP. These results suggest that in CHO cells: (i) the major route of mutagenic activation of 1- and 3-nitro-BaP involves S9-generated derivatives of the trans-7,8 dihydrodiols, e.g. bay-region diol epoxides; (ii) reactive nitroarene oxides may contribute to mutation induction by 3-nitro-BaP; and (iii) metabolic routes involving trans-9,10-dihydrodiol formation result in detoxification. PMID- 3045487 TI - Chemically induced aneuploidy in female germ cells. AB - Female mice were dosed with the spindle poisons colcemid (5 mg/kg) or colchicine (2 mg/kg) or with a dihydropyridazinone (ICI 109,081; 5 mg/kg; which inhibits oocyte maturation) or with saline, the vehicle control. The dosed females (whose ovulation was synchronized using exogenous gonadotrophins) were mated with undosed males, 12 or 3 h prior to ovulation, coresponding to MI or MII stages in preovulatory oocytes. First-cleavage embryos derived from these matings were then analysed for numerical chromosome aberrations. There were no numerical aberrations in the embryos derived from control matings. Polyploid embryos were isolated from females dosed with colchicine. When females were dosed 3 h prior to ovulation (PO) 26% of the first-cleavage embryos were polyploid; in those dosed 12 h PO it was 100%. Colcemid and ICI 109,081 induced both polyploid and aneuploid embryos; with colcemid these aberrations were only observed in the embryos derived from oocytes of females dosed 3 h PO. At this sample time 4% of the embryos were polyploid and 24% were aneuploid. In contrast, the majority of aberrations induced by ICI 109,081 were observed in females dosed 12 h PO where 10% of the embryos were polyploid and 3% were aneuploid. PMID- 3045486 TI - Isolation of S9 fractions from mouse and rat with increased enzyme activities after repeated administration of cytochrome P-450 and P-448 inducers. AB - Cytochrome P-450 (cyt P-450), NADPH cytochrome P-450 reductase and various microsomal monooxygenase activities [e.g. aminopyrine N-demethylase, p nitroanisole O-demethylase, dinemorphan N-demethylase, ethoxycoumarin O deethylase and ethoxyresorufin O-deethylase (ERD)], were determined in hepatic post-mitochondrial supernatant from mice and rats. Experiments were performed on male and female animals treated with a combination of sodium phenobarbital and beta-naphthoflavone according to the standard protocol schedule for short-term genotoxicity testing. A second inductive treatment after 2, 3, 4 or 5 weeks was provided. The increase in cyt P-450 and in all enzymatic activities measured was enhanced in both species by a second induction treatment, particularly when given after 4 weeks. ERD activity was the only monooxygenase activity which was sex dependent, being more active in female than in male animals. To extend the biochemical data, experiments were performed with the proposed S9 fractions on styrene, which previously has proved difficult to detect in short-term in vitro mutagenicity tests. Using the new induction conditions positive results were obtained with the D7 strain of Saccharomyces cerevisiae. It was concluded that a simple pre-induction of the animals 3-4 weeks before the main induction treatment leads to a more active S9 fraction for in vitro genotoxicity studies. PMID- 3045488 TI - Sensitivity of polA mutants of Escherichia coli K-12 to ozone and radiations. AB - Different polA mutants of Escherichia coli K-12 were exposed to ozone, X-rays and UV radiation, in order to compare the role of the various enzymatic activities of DNA polymerase I in the repair of damage caused by these agents. As was the case with radiations, the polymerase-deficient mutants were the most sensitive to ozone, followed by the 5'-3' exonuclease-deficient mutants. The 3'-5' exonuclease activity of pol I appeared to play a minor role in the repair of damage induced by all these agents. PMID- 3045490 TI - Ampicillin-resistance, a background to the use of this bacterial genetic marker in the presence of mammalian material. PMID- 3045489 TI - Mutagenicity, clastogenicity and cytotoxicity of procarcinogens in a rat hepatoma cell line competent for xenobiotic metabolism. AB - The present studies were aimed at evaluating the suitability of the differentiated Reuber hepatoma cells H4IIEC3/G- for monitoring permanent damage to the DNA caused by hepatotrophic chemicals. First we determined the profile of xenobiotic metabolizing enzymes. The cells expressed various cytochrome P-450 dependent monooxygenases, UDP-glucuronosyl-, phenol sulpho- and glutathione S transferase, cytochrome c (P-450) reductase and carboxylesterases. We then established the conditions for genotoxicity testing in H4IIEC/G- cells. Induction of resistance against 6-thioguanine and appearance of micronuclei served as indicators for mutagenicity and clastogenicity, respectively. 6-Thioguanine resistant H4IIEC3/G- cells were phenotypically stable for at least 30 cell cycles; recovery of 6-thioguanine-resistant cells was not significantly affected by the number of cells seeded for mutant selection up to at least 10(6) cells/100 mm dish; expression time of chemically induced mutants was 12-15 days; a period of 24 h after treatment appeared to be sufficient to allow for the formation of micronuclei. Finally we tested the genotoxic effects of promutagens which are typically activated or inactivated in liver. Aflatoxin B1, N-nitrosodiethylamine and cyclophosphamide were genotoxic to H4IIEC3/G- cells at concentrations of 10 30 nM, 2-20 mM and 1 mM, respectively. N-Nitrosodimethylamine and benzo[a]pyrene were not or only weakly cytotoxic and genotoxic to the cells, but this appears most likely to be due to protective mechanisms rather than to lack of metabolic activation. The results indicate that differentiated hepatoma cells such as H4IIEC3/G- offer a means of studying the potential of chemicals for inducing permanent DNA damage in liver cells. PMID- 3045491 TI - Why study mechanisms of cell death? AB - Cell death plays a variety of important roles in biology, both physiological and pathological. Physiological cell death is an important part of development, cell turnover and various aspects of immunological defenses. Pathological cell death is an element of some of the most important diseases plaguing modern society. Recent studies on the mechanisms of calcium-mediated cell death, thought to be important for both physiological and pathological cell death, have indicated that it is a multi-step process in which the cells are active participants, triggering their own self-destruction. Such mechanisms suggest the potential for pharmacological intervention, either to prevent disease-induced tissue death or to remove unwanted tissues. PMID- 3045492 TI - The anatomy and pathophysiology of the microvasculature in different organs: relationship to vasculogenic necrosis and tissue damage. AB - This chapter presents the methodology as well as various applications of a silicone rubber (Microfil) prefusion technique for evaluation of the microcirculation. In the heart, Microfil perfusion has been used in conjunction with clearing techniques to elucidate normal anatomic relationships, to identify areas of myocardium at risk following vascular occlusion, and to observe dynamic events in the development of various cardiomyopathies. Advantages of the technique include in vivo applicability, ease of vascular filling, and ability to discriminate between adjacent vascular fields. Preliminary data from work in other organ systems including skeletal muscle, brain, liver, and lung are also presented. PMID- 3045494 TI - Patterns of cell death. AB - Cell death takes two distinct forms, necrosis and apoptosis. Necrosis is a degenerative phenomenon that follows irreversible injury. Apoptosis, in contrast, appears to be an active process requiring protein synthesis for its execution; it is implicated in physiological regulation of tissue size, and, where it occurs pathologically, a homeostatic role for the death is often evident. Morphologically, apoptosis involves condensation of the nuclear chromatin and cytoplasm, fragmentation of the nucleus, and budding of the whole cell to produce membrane-bounded bodies in which organelles are initially intact. These bodies are disposed of by adjacent cells without inflammation. Biochemically, there is distinctive internucleosome cleavage of DNA in apoptosis, which is quite different from the random DNA degradation observed in necrosis. PMID- 3045493 TI - Metabolic processes leading to myocardial cell death. PMID- 3045495 TI - Pathogenesis of experimental alcoholic liver disease in the rat. PMID- 3045497 TI - Cell death in developing systems. AB - The so called 'normal', 'physiological' or 'programmed' cell death constitutes now a major field of developmental and cell biology. In the present article we review recent information on the biological significance of cell death in development. Special attention is paid to the range of techniques available for studies on cell death, the role of cell death during normal and abnormal development and to the mechanisms controlling cell death, including epigenetic and genetic factors. PMID- 3045496 TI - Ischemic bowel necrosis induced by platelet-activating factor: an experimental model. PMID- 3045498 TI - Anatomy and quantification of myocardial cell death. AB - Irreversible damage of the myocardial cells may show different morphologic aspects in relation to the type of dysfunction of their contraction-relaxation cycle. Attenuation of the muscle fibers with elongation of the sarcomeres and nuclei are the earliest modifications (systolic paradoxical bulging and stretching by the intraventricular pressure) when the myocells stop their function in irreversible relaxation. This 'atonic' death is pathognomonic of myocardial infarction (infarct or coagulation necrosis) and the lesion evolves with typical structural changes. An opposite and entirely different morphologic pattern is seen in the 'tetanic' death in which the myocardial cells arrest in irreversible contraction (coagulative myocytolysis or contraction band necrosis). Segmental (paradiscal bands) or pancellular hypercontraction with extreme shortening of the sarcomeres and subsequent myofibrillar rhexis alternated with irregular cross band formations (holocytic bands) are characteristic of this necrosis seen in numerous human and experimental conditions and specific of catecholamine toxicity. The third type of damage is observed in low output syndromes in which increasing edematous vacuolization and disappearance of the myofibrils (colliquative myocytolysis) are the main structural alterations. They are suggestive of progressive functional reduction leading to dilatative insufficiency ('failing' death). These clear-cut morphofunctional patterns indicate distinctive biochemical impairments and pathogenesis. In particular their frequent presence in and possible association with the different aspects of the ischemic heart disease presuppose other non-ischemic mechanisms responsible for complications and death in this modern epidemic. PMID- 3045499 TI - Expression and mutagenesis of the regulatory subunit of cAMP-dependent protein kinase in Escherichia coli. PMID- 3045500 TI - Preterm delivery of low birthweight infants. PMID- 3045501 TI - So you want to use computer assisted learning? PMID- 3045503 TI - [Changes in the energy status of synchronous Escherichia coli cultures during aerobic-anaerobic transitions]. AB - The behaviour of energy and growth parameters for a synchronous Escherichia coli culture was studied in the course of aerobic-anaerobic transitions induced by either reversible or constant cessation of aeration at different phases of the cell cycle. The intracellular concentrations of ATP, ADP and AMP were assayed using a technique developed by the authors and based on thin-layer chromatography of adenyl nucleotide dansyl derivatives. Prolonged anaerobiosis led to profound alterations in the energy status which diminished the energy flux in constructional processes and preserved the energy resources for maintenance needs. The response of the energy parameters to a transitory anaerobic shock depended on the phase of synchronous growth and, apparently, on the ratio between the rates of reactions supplying and consuming the energy, which underwent changes in the cell cycle. PMID- 3045502 TI - [Potential use of the monosaccharide composition of bacteria for their identification]. AB - The monosaccharide composition of cell hydrolysates can be used as a criterion for the chemical differentiation of gram-positive bacteria. The monosaccharide composition of six bacterial species belonging to the genus Bacillus has been determined using gas chromatography, mass spectrometry and computers. The qualitative composition was similar, glucose, galactose, ribose and glucosamine being the main components in all of the species. Some Bacillus species differed in their minor components. Although the monosaccharide composition appeared to be homogeneous, bacteria can be identified in terms of their carbohydrate profile using computers. To this end, the monosaccharide composition of bacterial cells is represented as a two-dimensional data file including the qualitative composition of components and the quantity of each component. PMID- 3045504 TI - [Lytic activity of Streptomyces levoris in relation to different bacterial groups]. PMID- 3045505 TI - The enigma of preterm birth. PMID- 3045506 TI - Who needs botulinum toxin? PMID- 3045507 TI - Cleveland--1980s; Vienna--1930s. PMID- 3045508 TI - [Kidney stones. From the stone drill to pain-free stone destruction]. PMID- 3045509 TI - Choice of cholesterol-lowering drugs. PMID- 3045511 TI - [Characteristics of the anabolic processes of the pancreas in trichinelliasis of various degrees of severity]. PMID- 3045510 TI - [Clinical characteristics of pseudotuberculosis in the presence of Opisthorchis infestation]. PMID- 3045512 TI - [Detection of IgE antibodies to ascarid antigens by a chemiluminescence method]. PMID- 3045513 TI - [Clinico-epidemiological characteristics of malaria imported into Donetsk Province (1979-1986)]. PMID- 3045514 TI - [Development of methods for the absolute count of the population density of pasture ixodid ticks in open landscapes. 2. An attempt at direct trapping of hungry larvae and nymphs of Dermacentor nuttalli Olen]. PMID- 3045515 TI - [Routes of entry into reservoirs, of diphyllobothriid eggs and their role in forming and maintaining foci of diphyllobothriasis]. PMID- 3045519 TI - [Cytogenetic evaluation of subgingival lesions: fibromatous, angiomatous, giant cell and granular-cell (so-called epulis)]. PMID- 3045520 TI - [The mouth and teeth in the "Canones de Medicina Solidorum" of Giorgio Baglivi]. PMID- 3045518 TI - Defense mechanisms involving Fc-dependent functions of immunoglobulin A and their subversion by bacterial immunoglobulin A proteases. PMID- 3045521 TI - Coping with war: an oral history of United States Army flight nurses of World War II. PMID- 3045522 TI - Exercise-induced asthma. AB - Though exercise-induced asthma (EIA) has been recognized for centuries, its characteristics, standardized testing, and pharmacologic management have been clarified only in the last two decades. Controversy continues concerning etiology; whether or not cold air, hypertonic and hypotonic bronchial challenges involve the same mechanism(s); and the incidence and clinical significance of late phase reactions. Aerosolized adrenergic agents such as albuterol or terbutaline, when administered prior to exercise, are usually effective in preventing EIA. Theophylline varies in effectiveness from subject-to-subject as does cromolyn sodium. Other agents such as H-1 antihistamines, ipratropium bromide, calcium channel blockers and adrenocorticosteroids are less effective when used alone, but may be useful when used in association with the more potent drugs. PMID- 3045517 TI - Mechanisms of gene regulation in the general control of amino acid biosynthesis in Saccharomyces cerevisiae. PMID- 3045524 TI - [Physiopathology of gastroesophageal reflux in pediatric patients]. PMID- 3045523 TI - Pathogenetic mechanisms of exercise-induced asthma and the refractory period. AB - Exercise is a powerful stimulus to the development of asthma. In most asthmatic subjects the airways obstruction recovers spontaneously within 60 minutes, but in some subjects there is more prolonged airflow obstruction which requires bronchodilator treatment. Approximately 40-50% of subjects with EIA will show a refractory period of two to four hours after an initial exercise task, during which time an identical exercise task will evoke significantly less (less than 50%) bronchoconstriction. In some patients, particularly children, EIA will be followed three to nine hours later by a further episode of bronchospasm, termed the late asthmatic response. There remains considerable debate about the pathogenesis of EIA the refractory period and the late asthmatic response. PMID- 3045526 TI - [Ultrasonographic detection of metastatic axillary lymph nodes in breast cancer]. PMID- 3045525 TI - [Cholecystolithiasis causing Mirizzi's syndrome with a rare anomaly of the extrahepatic bile duct in a child. Report of a case]. AB - A 14 year-old girl, who was admitted to our hospital due to increasing jaundice, intermittent right upper quadrant pain and fever, underwent operation based on the diagnosis of the Mirizzi's syndrome. Calculi filled the cystic duct and compressed the right hepatic duct. The right and left hepatic ducts lay closer to the duodenum than usual. The operation was limited to cholecystectomy alone. The calculi showed laminar cut surfaces and were composed of bilirubin lime. In addition to reporting our case, the literature dealing with this particular entity is reviewed. PMID- 3045527 TI - Nerve-muscle interrelationships in mammalian skeletal muscle regeneration. PMID- 3045528 TI - Detection in a murine T lymphoma of a lipid-like factor that inhibits cell growth. AB - In this paper we show that MCG3 cells, a murine T lymphoma, contain a factor(s) that inhibits the proliferation of cells of different histological origin. The lack of sensitivity of this cell proliferation-inhibiting factor (CPIF) to the treatment with proteolytic enzymes and its solubility in organic solvent demonstrated that it is a lipid-like substance. Separation by thin-layer chromatography showed it migrates before the prostaglandins with activity on cell proliferation. CPIF activity was reversible and more intense on bone marrow cells than on tumor cells, suggesting that it can play a role in cell growth regulation. PMID- 3045530 TI - [The role of bacteriophages in the study of the architectonics of the bacterial cell wall]. AB - Adsorption of bacteriophages on the surface of microbial cell is the first link in the chain of events leading to the development of bacteriophage infection. Adsorption is based on the presence of appropriate receptors of the various chemical nature. All the receptors are of significance for cellular metabolism. Some of them have the pathogenicity functions and antigenic properties. In course of bacterial dissociation some receptors disappear and others become available for bacteriophages. It is concluded, with a number of examples supporting the conclusion, that bacteriophages may be used in elucidation of cellular wall characteristics and architectonics as well as for the direct selection of mutant clones. The conclusion is also valid for bacteriocins, although, their chemical nature and location of receptors are not studied enough. PMID- 3045529 TI - Kinetics and location of bone marrow cell graft rejection by irradiated mice. AB - Natural killer (NK) cells were eliminated with rabbit anti-Asialo GM1 (anti ASGM1) serum to test the kinetics and location of bone marrow cell (BMC) rejection. Anti-ASGM1 serum was injected intravenously in mice at various times before or after irradiation (8.6 Gy) and transfer of parental-strain or allogeneic BMC. Growth of BMC was determined by measuring splenic 5-iodo-2' deoxyuridine-125I incorporation 5 days after cell transfer. Anti-ASGM1 serum weakened hybrid resistance even if injected intravenously as late as 24 h post BMC transfer and even in recipients injected with polyinosinic:polycytidylic acid so as to boost NK activity. If regenerating spleen cells (higher rate of cell cycling) were used as donor cells instead of BMC, the length of time required for rejection was unaffected. Anti-ASGM1 serum injected intravenously rapidly inhibited splenic NK activity and lung clearance of YAC-1 tumor cells, but when injected intratracheally, it only inhibited lung NK activity. Thus, BMC rejection occurs in the hematopoietic tissue and requires at least 24 h. PMID- 3045531 TI - SOS independent survival against mitomycin C induced lethality in a rifampicin nalidixic acid-resistant mutant of Escherichia coli. AB - A combination of specific rifampicin-resistant (rpoB87) and nalidixic acid resistant (gyrA87) mutations results in a marked increase in the survival of Escherichia coli against mitomycin C-induced lethality in mutants defective for SOS induction and excision repair. Although the response does not seem to be obligatorily dependent upon the RecA protein, the efficiency is markedly increased in its presence, even in a conventionally inactive form. This response is not elicited against lethality due to ultraviolet radiation or N-methyl-N' nitro-N-nitrosoguanidine exposure. The combination of rpoB87 and gyrA87 mutations also greatly alleviates post-mitomycin C degradation of DNA under SOS non inducible conditions. It is proposed that the rpoB subunit of RNA polymerase and gyrA subunit of DNA gyrase could participate in the repair of certain types of DNA damage, such as cross-links, in a mode independent of SOS-regulated excision repair and post-replication repair. PMID- 3045516 TI - Nitrate respiration in relation to facultative metabolism in enterobacteria. PMID- 3045533 TI - The mutagenic activity of gastric juice. AB - The mutagenic activity of fasting gastric juice was assessed in 123 patients including 18 with normal endoscopic findings, 53 peptic ulceration, 9 gastric cancer, 12 pernicious anaemia and 31 patients who had undergone peptic ulcer surgery in the past. Significant mutagenic activity was detected in 96 (78%). Marked variations in mutagenic activity were noted both within and between the patient groups and no significant differences were detected. No correlation was found between mutagenic activity and patient age or sex, gastric pH, bile acid concentrations or bacterial counts, intestinal metaplasia on gastric mucosal biopsy, or intragastric nitrite. About 30% of gastric juice samples showed evidence of a cytotoxic activity towards the Salmonella tester strains in the mutation assay. Preliminary studies on other body fluids showed the presence of significant mutagenic activity in fasting saliva, bile and plasma. These findings demonstrate widespread human exposure to potentially genotoxic substances. PMID- 3045532 TI - The induction of rho- mutants by UV or gamma-rays is independent of the nuclear recombinational repair pathway in Saccharomyces cerevisiae. AB - In order to discover whether the nuclear recombinational repair pathway also acts on lesions induced in mitochondrial DNA (mtDNA), the possible role of the RAD50, 51, -52, -55 and -56 genes on the induction of rho- mutants by radiations was studied. Such induction appeared to be independent of this pathway. Nevertheless, an efficient induction of respiration-deficient mutants was observed in gamma irradiated rad52 diploids. We demonstrate that these mutants do not result from a lack of mtDNA repair, but from chromosome losses induced by gamma-rays. Such an impairment of the respiratory ability of diploids by chromosome losses was effectively observed in the aneuploid progeny of unirradiated RAD+ cdc6 diploids incubated at the restrictive temperature. PMID- 3045535 TI - Genetic toxicology evaluation of commercial beers. I. Development of procedures for the preparation of extracts from commercial beer products. AB - Commercial beer was subjected to an investigation in order to establish standard conditions for preparing organic solvent extracts to be used in short-term genetic screening assays. Test samples for use in the evaluation were prepared by mixing several brands of commercially available beer into a composite pool which was then spiked with the mutagen, 2-nitrofluorene. The composite sample was then concentrated using varying ratios of beer to XAD-2 resin in a 1.5 cm X 30 cm column. Dry-weight analyses indicated that significant amounts of residue could be trapped by XAD-2 resin. Columns were sequentially eluted by methylene chloride, acetone and methanol followed by evaporation of the solvents under nitrogen gas. Residues from commercial products were not mutagenic, but mutagenic activity could be detected in residues from spiked beer, yielding nearly 90% of the expected biological activity in S. typhimurium TA98. A standard method amenable to processing large volumes of beer products was devised for application to other projects. PMID- 3045534 TI - Mutagenicity of aromatic glycidyl ethers with Salmonella. AB - 6 aromatic glycidyl ethers containing naphthyl, biphenyl or benzylphenyl substituents were synthesized. These epoxides together with the commercially available compounds 2-biphenylyl glycidyl ether were examined for dose mutagenicity relationships using the plate incorporation Ames test with Salmonella typhimurium strains TA100 and TA1535. Structure-mutagenicity relationships were further examined for these compounds and 3 phenyl glycidyl ethers by concurrent testing at a single dose with strain TA100. Meaningful correlations could not be established for the mutagenicity of these epoxides to their molecular volumes, partition values, nor to their reactivities with the model nucleophile, 4-(4-nitrobenzyl) pyridine. However, it was noted that increased conjugated aromatic unsaturation with its resulting planarity led to increased mutagenicity and that this effect decreased when it was further removed from the epoxide moiety. PMID- 3045536 TI - Genetic toxicology evaluation of commercial beers. II. Mutagenic activity of commercial beer products in Salmonella typhimurium strains TA98, TA100 and TA102. AB - 5 concentrated extracts of commercial beers were prepared using XAD-2 resin. The residues were subjected to evaluation for mutagenic activity in Salmonella typhimurium strains TA98, TA100 and TA102. The tests were conducted using preincubation protocols including provisions for S9 metabolic activation. Although the extracts did produce moderate toxicity to the Salmonella organisms used in the assays, none of the residues were found to induce mutation up to their maximum testable concentrations. PMID- 3045537 TI - Induction of chromosomal aberrations in rat bone marrow cells and mutations in Salmonella typhimurium by benz[a]anthracene derivatives. AB - Benz[a]anthracene (BA) and its derivatives containing methyl and/or ethyl groups in the 7 and/or 12 positions were tested for their ability to induce chromosome aberrations (CA) in rat bone marrow cells and for their mutagenicity to Salmonella typhimurium TA100 or TA98. The incidence of aberrant cells induced by the BA derivatives, given in lipid emulsion as a single-pulse dose of 50 mg/kg body weight into the caudal vein, was in the order: DMBA greater than EMBA greater than MEBA greater than other BA derivatives = control. The alkyl groups, at least 1 methyl group, at the 7 and 12 positions of BA seemed to be necessary to induce CA, although DEBA having ethyl groups at both the 7 and 12 positions of BA did not induce CA. DMBA or EMBA induced not only gaps and breaks but also exchanges and multiple CA, while the CA induced by other BA derivatives consisted of only gaps and breaks. 7MBA and 12MBA which exhibit carcinogenic activity intermediate between that of DMBA and BA induced few CA in the present system. However, the correlation coefficient between the logarithm incidence of aberrant cells and the carcinogenicity index calculated from the data of 9 BA derivatives including both 7MBA and 12MBA was 0.792. The relative mutagenicities of the BA derivatives with TA100 in the presence of hepatic S9 from polychlorinated biphenyl (PCB)-treated rats were in the order: BA greater than 7MBA greater than DMBA greater than 12MBA greater than 7EBA greater than EMBA greater than MEBA greater than 12EBA = DEBA = control. The results with TA98 were essentially the same as those with TA100. The results with TA100 in the presence of hepatic S9 from phenobarbital (PB)-treated rats were in the order: DMBA greater than 12MBA greater than 7MBA greater than 7EBA greater than BA greater than EMBA = MEBA greater than 12EBA = DEBA = control. These findings reveal no obvious relation between the mutagenic activities of the BA derivatives with the PCB-S9 or PB-S9 activating systems and their capacities to induce CA or their reported carcinogenicities. The incidence of CA induced by the dihydrodiols implicated as the metabolic precursors of the active diol epoxide metabolites of several of these BA derivatives was also tested. BA 3,4-dihydrodiol, like BA itself, induced few CA. However, the corresponding dihydrodiols of DMBA, 12MBA and 7MBA, induced relatively high levels of CA.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3045538 TI - Mutagen formed from tryptophan reacted with sodium nitrite in acidic solution. AB - The reaction products from L-tryptophan treated with nitrite under acidic conditions were investigated for mutagenic activity with the Salmonella typhimurium his reversion assay and for DNA-damaging activity using the rec assay. The diethyl ether extract of the reaction mixture showed 8 spots on thin layer chromatography (TLC). One compound from the TLC had high mutagenic activity for TA98 without S9 mix, with little DNA-damaging activity. The mutagen was purified and identified by instrumental analysis as 2-hydroxy-(1-N nitrosoindole)propionic acid (NIHP). The mutagenic activity of NIHP was determined by the induced mutation frequency method; the induced mutation frequency was about 19.2 X 10(-5) at a dose level of 160 micrograms/plate. PMID- 3045539 TI - Isolation and partial characterisation of a 26 kilodalton antigen from Plasmodium falciparum recognised by an inhibitory monoclonal antibody. AB - A 26 kDa protein, present in trophozoites and schizonts of Plasmodium falciparum, has been identified as the target of a monoclonal antibody that weakly inhibits parasite growth in vitro. The antigen has been purified to homogeneity by immuno affinity chromatography and electrophoresis. The sequence of 19 amino acids at the N-terminus of the protein has been determined. PMID- 3045540 TI - A tubulin-related 55 kilodalton surface antigen recognized by different Trypanosoma cruzi stage-specific monoclonal antibodies from infected mice. AB - Thirteen monoclonal antibodies (MAbs) specific for the membrane of live Trypanosoma cruzi have been obtained from BALB/c infected mice. Most of them had greater avidity for intact than for disrupted parasites. According to the staining by indirect immunofluorescence of the different live developmental stages of the parasite the MAbs could be divided into several groups. Three of them were trypomastigote specific, one amastigote-specific and two epimastigote specific. The rest reacted with either all stage forms or with various combinations of the different stages. However, despite the fact that they seemed to correspond to stage-specific antibodies, ten of them reacted with the same 55/50 kDa antigen by immunoblotting. Similarly, a 55 kDa protein was immunoprecipitated from these MAbs. By contrast, a single band or a dimer of about 25 kDa was the predominant antigen(s) immunoprecipitated by the same MAbs in absence of protease inhibitors. This smaller protein may arise from proteolysis of the 55 kDa band. This protein is related to tubulin since tubulin (a 55 kDa protein) but not other cytoskeleton proteins blocked the binding of these MAbs to T. cruzi, and some MAbs react with pig alpha-tubulin by immunoblotting. PMID- 3045541 TI - Characterization of a family of rhoptry proteins of Toxoplasma gondii. AB - A monoclonal antibody (McAb 4A7) raised against a rhoptry enriched subcellular fraction of Toxoplasma gondii tachyzoites reacted in immunofluorescence studies with rod-like organelles located in the anterior part of the organisms and gave specific labeling of rhoptries in immunoelectron microscopy. On immunoblots, two major proteins of 55 and 60 kDa were identified by McAb 4A7. Similar results were obtained both by immunodetection and immunoblotting with tachyzoites, bradyzoites and sporozoites. Pulse chase analysis of [35S]methionine labeled tachyzoites demonstrated that the 55 and 60 kDa rhoptry proteins derived from a 66-68 kDa doublet which was processed approximately 30 min after biosynthesis. Two other monoclonal antibodies (McAb 2F8, McAb 2H3) respectively specific for rhoptry proteins of 55 kDa having a 66 kDa precursor and 60 kDa having a 68 kDa precursor were also obtained; we suggest that they recognize separately the two components of the 55-60 kDa rhoptry protein family of Toxoplasma. PMID- 3045542 TI - Alterations in intracellular calcium following infection of human endothelial cells with Trypanosoma cruzi. AB - Trypanosoma cruzi infection in cultured human umbilical vein endothelial cells increased basal cellular calcium levels from 55 to 110 nM, as monitored with the fluorescent probe, fura-2. It also influenced intracellular calcium such that consistently higher total levels were observed in response to bradykinin, angiotensin II and norepinephrine, as compared to similarly treated uninfected cells. However, bradykinin and angiotensin II-dependent increases in calcium, when considered as the absolute increment or fold elevation over basal, were significantly lower in infected endothelial cells. Infection also influenced changes in calcium levels due to agents that operate independently of plasma membrane receptors. In the presence of ionomycin, the magnitude and rate of rise of intracellular calcium were decreased; additionally the calcium peak was delayed and the subsequent decline slowed. Similar to the results with bradykinin and angiotensin II, infection decreased both the increment in and fold stimulation of intracellular calcium in response to ionomycin. In contrast, infection altered only the total calcium stimulated in response to oligomycin; neither the fold stimulation of, nor increment in intracellular calcium was affected. These results indicate that (1) infection by T. cruzi alters calcium homeostasis in endothelial cells under basal and stimulated conditions; (2) both receptor-dependent and receptor-independent mechanisms are affected by infection. The possible contribution of altered calcium homeostasis induced by T. cruzi in the pathogenesis of chagasic cardiomyopathy is considered. PMID- 3045543 TI - The 140/130/105 kilodalton protein complex in the rhoptries of Plasmodium falciparum consists of discrete polypeptides. AB - Four monoclonal antibodies (MAbs) recognise an antigen localised in the rhoptries of Plasmodium falciparum merozoites using both indirect immunofluorescence assay and immunoelectron microscopy with immunogold labeling. All MAbs immunoprecipitated bands at 140, 130 and 105 kDa from [35S]methionine-labeled parasites; however, one MAb immunoblotted only the 130 kDa protein and another MAb immunoblotted the 105 kDa protein. The affinity purified antigen complex consisted of proteins of 140, 130, 105 and 98 kDa. The individual proteins were subjected to peptide mapping with Staphylococcus aureus V8 protease; the 98 kDa protein was a degradation product of the 105 kDa protein and the 140, 130, and 105 kDa proteins were found to be unrelated. The antigen complex was synthesised at the mid trophozoite stage and was considered to be soluble as judged by release from mature schizonts by freeze/thaw lysis. One of the MAbs inhibited parasite growth and/or merozoite invasion of erythrocytes, in vitro, to a small but significant extent. PMID- 3045544 TI - Effect of recombinant human granulocyte-macrophage colony-stimulating factor on chemotherapy-induced myelosuppression. AB - An increase in the dose of chemotherapy enhances the response of many experimental and clinical cancers, but the extent of dose escalation is often limited by myelosuppression. In preliminary trials, recombinant human granulocyte macrophage colony-stimulating factor (rhGM-CSF) has augmented leukocyte numbers and function, but the optimal dose is not established. We treated 16 adults who had inoperable or metastatic sarcomas with escalating doses of rhGM-CSF before and immediately after a first cycle of chemotherapy (cycle 1) to assess hematologic response and toxicity. A second cycle of chemotherapy (cycle 2) was given without rhGM-CSF. RhGM-CSF was tolerated well at doses of 4 to 32 micrograms per kilogram of body weight per day. At 64 micrograms per kilogram per day, edema and thrombi around a central venous catheter developed in two of four patients. Leukocyte and granulocyte counts increased significantly during the rhGM-CSF infusion. Neutropenia after cycle 1 was significantly less severe and shorter in duration than after cycle 2 (P less than 0.01). Mean total leukocyte and platelet nadirs were 1.0 and 101 x 10(9) per liter for cycle 1 and 0.45 and 44 x 10(9) per liter for cycle 2 (P less than 0.01), and the median intervals from day 1 of chemotherapy to neutrophil recovery (greater than 0.500 x 10(9) per liter) were 15 and 19 days, respectively (P less than 0.01). The duration of neutropenia was 3.5 days with cycle 1 and 7.4 days with cycle 2 (P less than 0.01). We conclude that rhGM-CSF is tolerated well at doses up to 32 micrograms per kilogram per day and is biologically active in leukopenic patients. It merits further evaluation for the prevention of morbidity from chemotherapy. PMID- 3045546 TI - Prerenal failure: a deleterious shift from renal compensation to decompensation. PMID- 3045545 TI - Immunosuppression with azathioprine and prednisone in recent-onset insulin dependent diabetes mellitus. AB - We randomly assigned 46 patients (mean age, 11.7 years; range, 4.5 to 32.8) with newly diagnosed insulin-dependent diabetes mellitus within two weeks of beginning insulin to receive either corticosteroids for 10 weeks plus daily azathioprine for one year or no immunosuppressive therapy. Half the 20 immunosuppressed patients completing the one-year trial had satisfactory metabolic outcomes (hemoglobin A1c less than 6.8 percent; stimulated peak C peptide greater than 0.5 nmol per liter; insulin dose less than 0.4 U per kilogram of body weight per day) as compared with only 15 percent of the controls. Three of 20 immunosuppressed patients, but no controls, were insulin independent at one year. Two of these continue to receive azathioprine without insulin after more than 27 months of follow-up. The response to immunosuppression correlated with older age, better initial metabolic status, and lymphopenia (less than 1800 lymphocytes per cubic millimeter) resulting from immunosuppression. The side effects of azathioprine included vomiting in one patient and mild hair loss in several others. Prednisone use resulted in a transient cushingoid appearance, weight gain, and hyperglycemia. The growth rate remained normal in all patients. We conclude that early immunosuppression with short-term use of corticosteroids plus daily azathioprine can improve metabolic control in some patients with insulin dependent diabetes mellitus, but results from this unblinded study are preliminary and require further confirmation and long-term follow-up. PMID- 3045547 TI - Interhospital patient transfer. The case for informed consent. PMID- 3045548 TI - Steroids and "steroid-sparing" agents in asthma. PMID- 3045549 TI - Routine use of phenolized formalin in fixation of autopsy brain tissue to reduce risk of inadvertent transmission of Creutzfeldt-Jakob disease. PMID- 3045551 TI - Clinical implications of prostaglandin and thromboxane A2 formation (2). PMID- 3045550 TI - Clinical implications of prostaglandin and thromboxane A2 formation (1). PMID- 3045552 TI - Computer-derived protocol to predict myocardial infarction in patients with chest pain. PMID- 3045554 TI - Current concepts. Acute mountain sickness. PMID- 3045553 TI - Comparison of a high-carbohydrate diet with a high-monounsaturated-fat diet in patients with non-insulin-dependent diabetes mellitus. AB - We compared a high-carbohydrate diet with a high-fat diet (specifically, a diet high in monounsaturated fatty acids) for effects on glycemic control and plasma lipoproteins in 10 patients with non-insulin-dependent diabetes mellitus (NIDDM) receiving insulin therapy. The patients were randomly assigned to receive first one diet and then the other, each for 28 days, in a metabolic ward. In the high carbohydrate diet, 25 percent of the energy was in the form of fat and 60 percent in the form of carbohydrates (47 percent of the total energy was in the form of complex carbohydrates); the high-monounsaturated-fat diet was 50 percent fat (33 percent of the total energy in the form of monounsaturated fatty acids) and 35 percent carbohydrates. The two diets had the same amounts of simple carbohydrates and fiber. As compared with the high-carbohydrate diet, the high-monounsaturated fat diet resulted in lower mean plasma glucose levels and reduced insulin requirements, lower levels of plasma triglycerides and very-low-density lipoprotein cholesterol (lower by 25 and 35 percent, respectively; P less than 0.01), and higher levels of high-density lipoprotein (HDL) cholesterol (higher by 13 percent; P less than 0.005). Levels of total cholesterol and low-density lipoprotein (LDL) cholesterol did not differ significantly in patients on the two diets. These preliminary results suggest that partial replacement of complex carbohydrates with monounsaturated fatty acids in the diets of patients with NIDDM does not increase the level of LDL cholesterol and may improve glycemic control and the levels of plasma triglycerides and HDL cholesterol. PMID- 3045555 TI - Perspectives on physician-payment reform. The resource-based relative-value scale in context. PMID- 3045556 TI - Granulocyte-macrophage colony-stimulating factor and marrow transplantation. PMID- 3045557 TI - Results and policy implications of the resource-based relative-value study. AB - The resource-based relative-value scale (RBRVS) is a measure of relative levels of resource input expended when physicians produce services and procedures. It is a function of the physician's work input, the opportunity cost of specialty training, and the relative practice costs for each specialty. This paper presents resource-based relative values (RBRVs) for selected procedures of four major specialties--family practice, internal medicine, general surgery, and thoracic and cardiovascular surgery. We compare RBRVs with current charges and find several general patterns. Invasive procedures are typically compensated at more than double the rate of evaluation-and-management services, when both consume the same resource inputs. Imaging and laboratory procedures fall between invasive and evaluation-and-management services. We analyze the financial implications of the RBRVS by developing a simple model and simulating the effects of an RBRVS-based fee schedule on physicians' revenues in various specialties. We use Medicare data to perform the simulation under the "budget-neutral" assumption. Results show that an RBRVS-based fee schedule affects specialties differently. The average family practitioner could receive 60 percent more revenue from Medicare, whereas the average ophthalmologist could lose 40 percent of current revenues. The effects on other specialties fall between these two. PMID- 3045558 TI - Structure of protein solutions. Part 2. Solutions of total protein of yeast Saccharomyces cerevisiae. AB - Water systems formed by total protein of yeast Saccharomyces cerevisiae and model systems on the basis of unfractionated casein were studied by viscosimetry and electron microscopy. Methodologically, both methods offered a possibility of distinguishing several quantitatively different concentration intervals in protein solutions under different conditions. Viscosimetry gave the most informative results at the concentration intervals where the solutions approached the true ones, while electron microscopy did so when supermolecular formations prevailed. In comparing the data obtained by these two methods the following conclusions were drawn: At low temperatures (4-10 degrees C) yeast protein was in molecular-dissociated condition. Association in the system starts at 10 degrees C, and becomes enhanced with a rise in temperature. A temperature rise to 25-30 degrees C promotes a more compact packing of polypeptide chains; further heating leads to high-temperature denaturation. Lipids induce covering of polypeptide chains and prevent associations, thus leading to an increase in the system's thermostability. PMID- 3045559 TI - Crimean-Congo haemorrhagic fever. A protocol for control and containment in a health care facility--Part 2. PMID- 3045560 TI - Victoria Hospital Wynberg. A trip down memory lane. PMID- 3045561 TI - A history of surgical stapling. PMID- 3045562 TI - The molecular heterogeneity of protein kinase C and its implications for cellular regulation. AB - Protein kinase C is now known to be a large family of proteins, with multiple subspecies that have subtle individual enzymological characteristics. Some members of the family exhibit distinct patterns of tissue expression and intracellular localization; different kinases probably have distinct functions in the processing and modulation of a variety of physiological and pathological responses to external signals. PMID- 3045563 TI - A highly conserved amino-acid sequence in thrombospondin, properdin and in proteins from sporozoites and blood stages of a human malaria parasite. AB - As a consequence of gene cloning and DNA sequencing several gene families are emerging in the field of cell-cell recognition. These include immunoglobulins, integrins, certain extracellular glycoproteins and a family of functionally unrelated proteins which include factor B. We report here the cloning and sequencing of a gene from Plasmodium falciparum, coding for a protein we call thrombospondin related anonymous protein (TRAP), which shares certain sequence motifs common to other well-characterized proteins. The most significant homology is based around the sequence Trp-Ser-Pro-Cys-Ser-Val-Thr-Cys-Gly (WSPCSVTCG), present in three copies in region I of thrombospondin (TSP), six copies in properdin (P) and one copy in all the circumsporozoite (CS) proteins sequenced so far. TRAP also shares with certain extracellular glycoproteins, including TSP, the cell-recognition signal Arg-Gly-Asp (RGD), which has been shown to be crucial in the interaction of several extracellular glycoproteins with members of the integrin superfamily. Unlike the CS protein, TRAP is expressed during the erythrocytic stage of the parasite life cycle. PMID- 3045564 TI - Properdin, the terminal complement components, thrombospondin and the circumsporozoite protein of malaria parasites contain similar sequence motifs. AB - Properdin is a plasma glycoprotein which stabilizes the C3bnBb enzyme complex of the alternative pathway of the complement system. Unlike the classical pathway, which is initiated by interaction of C1q with the Fc regions of IgG or IgM antibodies in immune complexes, the alternative pathway can be directly activated via binding of C3b to surfaces of foreign organisms. The stabilized C3bnBbP complex activates components C3 and C5 resulting in opsonization of foreign material (via C3b) and assembly of the membrane attack complex (via C5b) on target cells. Therefore properdin greatly enhances complement-mediated clearance and inactivation mechanisms in both natural and acquired resistance to infection. This paper shows that the primary amino acid sequence of properdin is composed mainly of six repeating motifs, each of approximately 60 amino acids, and that similar sequences are found in thrombospondin, the circumsporozoite protein of malaria parasites and regions of the membrane-attack components of complement. These similarities may provide insight into the mechanisms by which parasites avoid host defences mediated by complement. PMID- 3045565 TI - The origin of mutants. AB - Nucleic acids are replicated with conspicuous fidelity. Infrequently, however, they undergo changes in sequence, and this process of change (mutation) generates the variability that allows evolution. As the result of studies of bacterial variation, it is now widely believed that mutations arise continuously and without any consideration for their utility. In this paper, we briefly review the source of this idea and then describe some experiments suggesting that cells may have mechanisms for choosing which mutations will occur. PMID- 3045566 TI - Genome linking with yeast artificial chromosomes. AB - The haploid genome of Caenorhabditis elegans consists of some 80 x 10(6) base pairs of DNA contained in six chromosomes. The large number of interesting loci that have been recognized by mutation, and the accuracy of the genetic map, mean that a physical map of the genome is highly desirable, because it will facilitate the molecular cloning of chosen loci. The first steps towards such a map used a fingerprinting method to link cosmid clones together. This approach reached its practical limit last year, when 90-95% of the genome had been cloned into 17,500 cosmids assembled into some 700 clusters (contigs), but the linking clones needed were either non-existent or extremely rare. Anticipating this, we had planned to link by physical means--probably by hybridization to NotI fragments separated by pulse field gel electrophoresis. NotI recognizes an eight base sequence of GC pairs; thus the fragments should be large enough to bridge regions that clone poorly in cosmids, and, with no selective step involved, would necessarily be fully representative. However, with the availability of a yeast artificial chromosome (YAC) vector, we decided to use this alternative source of large DNA fragments to obtain linkage. The technique involves the ligation of large (50 1,000 kilobase) genomic fragments into a vector that provides centromeric, telomeric and selective functions; the constructs are then introduced into Saccharomyces cerevisiae, and replicate in the same manner as the host chromosomes. PMID- 3045567 TI - James Watson to head NIH human genome project. PMID- 3045568 TI - Serotonin stimulates DNA synthesis in fibroblasts acting through 5-HT1B receptors coupled to a Gi-protein. AB - Growth factors can be divided into two classes which act through distinct signal transduction pathways. One class including epidermal growth factor, platelet derived growth factor and fibroblast growth factor activates receptor tyrosine kinases, and the second class, including thrombin, bombesin, bradykinin and vasopressin activates a phosphoinositide-specific phospholipase C through GTP binding proteins which can be inactivated by pertussis toxin. In Chinese hamster lung fibroblasts, thrombin-induced mitogenicity seems to correlate well with phospholipase C activation and both events are sensitive to pertussis toxin. Thrombin, like the other mitogens in this class, simultaneously inhibits adenylate cyclase. This involves an inhibitory G protein (Gi), a well established pertussis toxin substrate. The relative contributions of the two signalling pathways to mitogenicity has not been evaluated so far. We report here that the neurotransmitter serotonin (5-hydroxytryptamine), a contracting agent and mitogen for smooth muscle cells, activates phospholipase C, inhibits adenylate cyclase and stimulates DNA synthesis in fibroblasts. These events are sensitive to pertussis toxin. We show that the mitogenicity of 5-hydroxytryptamine can be uncoupled from phospholipase C activation that is mediated by 5-HT2 receptors, but correlates perfectly with inhibition of adenylate cyclase through 5-HT1B receptor. We propose that inhibition of adenylate cyclase or activation of an undefined effector system by Gi is important in 5-hydroxytryptamine induced DNA synthesis and contributes to the strong mitogenicity of the other members of this family of growth factors. PMID- 3045569 TI - [Food allergy in children]. PMID- 3045570 TI - [Louis Heijermans (1873-1938) and the origin of work and occupational medicine]. PMID- 3045571 TI - [Insulin in the balance]. PMID- 3045573 TI - Post-traumatic stress syndrome. PMID- 3045572 TI - [Gastroesophageal reflux and bronchospasm in adults]. PMID- 3045574 TI - The cost of nursing. PMID- 3045575 TI - Parkinson's disease. PMID- 3045576 TI - Never alone. PMID- 3045577 TI - Conveying calming care. PMID- 3045579 TI - A good sign. PMID- 3045578 TI - On the boil. PMID- 3045580 TI - Teenage wildlife. PMID- 3045581 TI - Freedom to roam. PMID- 3045583 TI - Career talk. PMID- 3045582 TI - Weaning intentions. PMID- 3045584 TI - Imperfect balance. PMID- 3045585 TI - Celtic takeover. Interview by Tim Rice. PMID- 3045586 TI - A double act a phone call away. Interview by Bronwen Jones. PMID- 3045587 TI - The cooler culprits. PMID- 3045588 TI - Head first. PMID- 3045589 TI - Unproductive labour. PMID- 3045590 TI - What's in a report? PMID- 3045591 TI - Supporting role. PMID- 3045592 TI - Homeward bound. PMID- 3045593 TI - Safety first. PMID- 3045594 TI - Equality health care. Interview by Tim Rice. PMID- 3045595 TI - Other people's children. Interview by Charlotte Alderman. PMID- 3045596 TI - On the other side. PMID- 3045597 TI - Wishing on a star. Interview by Charlotte Alderman. PMID- 3045598 TI - Mobilising health care. PMID- 3045599 TI - Blood glucose monitoring. PMID- 3045601 TI - Project outline. PMID- 3045602 TI - The fairer sex. PMID- 3045603 TI - Rights of students. PMID- 3045600 TI - Shock tactics. PMID- 3045605 TI - Grassroots commitment. Interview by Tim Rice. PMID- 3045606 TI - Fixing it. Interview by Tim Rice. PMID- 3045604 TI - Starting from scratch. PMID- 3045607 TI - A milestone in nursing. PMID- 3045608 TI - Trading places. PMID- 3045609 TI - Apply within. PMID- 3045610 TI - Abortion. PMID- 3045611 TI - A case for debate. PMID- 3045612 TI - Final recognition. PMID- 3045613 TI - Radical intervention. Interview by Tim Rice. PMID- 3045614 TI - Triple time. Interview by Bronwen Jones. PMID- 3045615 TI - A respected speciality. PMID- 3045616 TI - How to find a publisher. PMID- 3045617 TI - The power over death. PMID- 3045618 TI - Euthanasia: the students' debate. PMID- 3045619 TI - Using restraint. PMID- 3045620 TI - Raynaud's syndrome. PMID- 3045621 TI - Coeliac disease. PMID- 3045622 TI - Vital vitamins. PMID- 3045623 TI - Body language: non-verbal communication and the nurse. PMID- 3045624 TI - Colorectal cancer. PMID- 3045625 TI - Browned off. PMID- 3045627 TI - The great divide. PMID- 3045626 TI - A remedy for a sick NHS. PMID- 3045628 TI - Outrage over a secret service. PMID- 3045629 TI - Room at the top. Interview by Tim Rice. PMID- 3045630 TI - Love and bullets. Interview by Tim Rice. PMID- 3045631 TI - How to write effectively. PMID- 3045632 TI - Safe and sound. PMID- 3045633 TI - Support for the dying. Interview by Charlotte Alderman. PMID- 3045634 TI - Renal dialysis. PMID- 3045635 TI - Benign breast disease. PMID- 3045637 TI - Tenosynovitis. PMID- 3045636 TI - Computers in nursing--an expensive paperweight? PMID- 3045638 TI - Managing epilepsy. PMID- 3045639 TI - Psychiatric nursing. PMID- 3045640 TI - Mistaken identity. PMID- 3045641 TI - Forty years on. PMID- 3045642 TI - Voicing concern. PMID- 3045643 TI - A force to be reckoned with. Interview by Tim Rice. PMID- 3045644 TI - Star treatment. Interview by Tim Rice. PMID- 3045645 TI - AIDS forum. PMID- 3045646 TI - Free agent. PMID- 3045647 TI - How to speak effectively. PMID- 3045648 TI - Profits of fear. PMID- 3045649 TI - Historical perspective. PMID- 3045650 TI - Nit picking. PMID- 3045651 TI - Allergic--or not? PMID- 3045653 TI - Psychiatric nursing--current practice. PMID- 3045652 TI - Discharge planning. PMID- 3045654 TI - Opportunities for enrolled nurses. PMID- 3045655 TI - Honest sweat. PMID- 3045656 TI - Giving good counsel. Interview by Tim Rice. PMID- 3045657 TI - Making music. Interview by Tim Rice. PMID- 3045659 TI - Patients affected by changed locations. PMID- 3045660 TI - Nursing and AIDS. 1. PMID- 3045658 TI - A supporting role. PMID- 3045661 TI - Poisons. PMID- 3045663 TI - The treatment of choice. PMID- 3045662 TI - Informed decisions. PMID- 3045664 TI - Sleepless nights. PMID- 3045665 TI - Bad for your health. PMID- 3045666 TI - A global context. Interview by Tim Rice. PMID- 3045667 TI - How to present a manuscript. PMID- 3045669 TI - Working in refugee health. PMID- 3045668 TI - A capital adviser. Interview by Tim Rice. PMID- 3045670 TI - Sleep of the just. PMID- 3045671 TI - From hospice to respice. PMID- 3045672 TI - Body heat. PMID- 3045673 TI - Community care for the mentally handicapped. PMID- 3045674 TI - Nursing and AIDS. PMID- 3045676 TI - The idea and the reality. Interview by Tim Rice. PMID- 3045675 TI - How to generate self-confidence. PMID- 3045677 TI - A prize winner. Interview by Tim Rice. PMID- 3045678 TI - Breast cancer--evolving concepts of treatment. PMID- 3045679 TI - The University of Nebraska Hospital at Omaha. By J. Jay Keegan, 1927. PMID- 3045680 TI - [The personality of bipolar manic-depressive patients]. PMID- 3045681 TI - Cells and mediators in glomerulonephritis. PMID- 3045682 TI - Apparent recurrence of hemolytic uremic syndrome in azathioprine-treated allograft recipients. AB - The present study describes 3 adult patients with hemolytic uremic syndrome (HUS) leading to chronic renal failure. Following renal transplantation, the 3 patients developed microangiopathic hemolytic anemia associated with acute rejection crises and graft failure. In 2 patients the renal histology of the transplanted kidney was consistent with HUS. It is concluded that, in adult patients, HUS may recur after renal transplantation and contribute to renal graft failure. PMID- 3045683 TI - Immunoglobulin A nephropathy and ankylosing spondylitis. Report of two patients in Taiwan and review of the literature. AB - We describe 2 patients with immunoglobulin A (IgA) nephropathy in association with ankylosing spondylitis. Renal biopsy demonstrated mesangial proliferative glomerulonephritis with prominent IgA, C3c, and less intense properdin deposition in the glomeruli. Intraglomerular clumps of virus-like particles were also observed in the kidney sample from one patient (case 2), who had an abnormal liver function. Our findings support the hypothesis that here is a possible common pathogenesis responsible for the concurrence of both IgA nephropathy and ankylosing spondylitis. PMID- 3045684 TI - Irreversible dementia following cyclosporin therapy in a renal transplant patient. PMID- 3045685 TI - Sublingual captopril in hypertensive attacks in patients on hemodialysis. PMID- 3045686 TI - Steroid hormones and the brain: linking "nature" and "nurture". AB - Contrary to earlier belief, the genetic constitution of each cell of the body ("nature") is subject to modulation by environmental factors ("nurture") which act throughout the life of the organism to shape the individual characteristics. The nervous system adapts and changes with the environment that the organism experiences through genomic activity controlled by chemical messengers from other nerve cells and from endocrine secretions. The nervous system expresses receptors for a number of circulating hormones, and the location of these hormone receptors has revealed a great deal about the neuroanatomy of neuroendocrine and behavioral control processes. The brain controls the endocrine system through the hypothalamus and pituitary gland, and it responds to circulating hormones throughout each stage of life. These effects begin during early development (eg., sexual differentiation of the brain; effects of maternal or neonatal stress). They continue in adult life in response to cyclic events (eg., season of year; time of day, controlling reproduction and daily activity-sleep rhythms of behavior); and they also include the behavior of other animals which alters hormone output. Hormones also operate during the aging process and under conditions which induce neural damage such as hypoxia and stress. This overview summarizes involvement of steroid hormones of gonads and adrenals in many of these processes and also examines the features of the genomic activity which is modified by these hormones. This area of research is fruitful because it brings together molecular, anatomical, physiological and behavioral approaches in an attempt to understand the long-term plasticity of the nervous system. PMID- 3045688 TI - ["One-knot" microvascular anastomosis using glutide as an external splint- experimental arterio-arterial anastomosis in rats]. AB - Experimental microvascular anastomosis using a glutide copolymer (lactide: glycolide = 80: 20) as an external splint was undertaken in rats between the left and the right carotid arteries. Both arteries were dissected free over a 1-cm length, the left carotid artery was transected at the cranial end, and the right carotid artery was cut at the caudal end. The left carotid artery was then introduced into a glutide pipe-splint. The arterial wall was turned back 180 degrees over the edge of the splint. The reflected part of the artery and the glutide were covered with the freed-up right carotid artery. One stitch was made around the two arteries and the glutide in a manner similar to that of binding a barrel with steel wire. The "One-knot anastomosis" was then complete. We call this type of anastomosis "antegrade anastomosis". If, on the other hand, the right carotid artery is introduced into the pipe, turned back at the edge of the glutide, covered with the left carotid artery and secured with one stitch, the technique is known as "retrograde anastomosis". The patency rates of the resulting vessels were as follows: antegrade anastomosis, 83% (15/18); and retrograde anastomosis, 92% (23/25); so that the average patency rate was 88% (38/43). We measured the anastomosing time that is the time between the transection of one of the two arteries and the completion of the anastomosis. The average anastomosing time was 21 minutes for antegrade anastomosis and 14 minutes for retrograde anastomosis. But the 'pure anastomosing time' (the time taken to connect the two already prepared arteries) was 2-3 minutes. We believe that one knot anastomosis technique for future clinical application is promising. PMID- 3045687 TI - Cell membranes: the electromagnetic environment and cancer promotion. AB - Use of weak electromagnetic fields to study the sequence and energetics of events that couple humoral stimuli from surface receptor sites to the cell interior has identified cell membranes as a primary site of interaction with these low frequency fields. Field modulation of cell surface chemical events indicates a major amplification of initial weak triggers associated with binding of hormones, antibodies and neurotransmitters to their specific binding sites. Calcium ions play a key role in this stimulus amplification, probably through highly cooperative alterations in binding to surface glycoproteins, with spreading waves of altered calcium binding across the membrane surface. Protein particles spanning the cell membrane form pathways for signaling and energy transfer. Fields millions of times weaker than the membrane potential gradient of 10(5) V/cm modulate cell responses to surface stimulating molecules. The evidence supports nonlinear, nonequilibrium processes at critical steps in transmembrane signal coupling. Powerful cancer-promoting phorbol esters act at cell membranes to stimulate ornithine decarboxylase which is essential for cell growth and DNA synthesis. This response is enhanced by weak microwave fields, also acting at cell membranes. PMID- 3045689 TI - Gnostic micronets of cortical neurons. PMID- 3045691 TI - Trends and perspectives in the development of the physiology of higher nervous activity. PMID- 3045690 TI - Interaction of brain macrostructures in the behavior organization process. PMID- 3045692 TI - Autoimmune paraneoplastic cerebellar degeneration: ultrastructural localization of antibody-binding sites in Purkinje cells. AB - Sera from three of four patients with paraneoplastic cerebellar degeneration (PCD) associated with gynecologic cancer had antibodies that stained the cytoplasm of Purkinje cells in a characteristic discrete and coarsely granular pattern. No such antibodies were found in PCD patients with small cell cancer of the lung, in patients with cerebellar degeneration without cancer, in nonneurologic patients with small cell carcinoma or gynecologic cancer, or in normal subjects. Immunoelectron microscopy revealed that the antibodies of PCD bound to clusters of ribosomes, granular endoplasmic reticulum, and the trans face of the vesicles of the Golgi complex in Purkinje cells. Immunostaining was localized in orderly arrays of stacked parallel cisternae of the endoplasmic reticulum in the perikaryon and dendritic processes. This pattern suggested that at least one autoantigen of PCD may be a glycoprotein specific cerebellar tissue that is associated with the endoplasmic reticulum. Some patches of Purkinje plasma membrane also were immunostained. PMID- 3045694 TI - [Intraoperative fine-needle biopsy of the pancreas. Analysis of 72 cases]. PMID- 3045693 TI - Creutzfeldt-Jakob disease: correlation of MRI and neuropathologic findings. AB - In a patient with Creutzfeldt-Jakob Disease (CJD), MRI showed increased signal intensity in striatum, thalamus, and cerebral cortex in images obtained with TR 1,600 msec, and TE 35 and 70 msec. In postmortem examination, all affected areas showed the hallmarks of CJD, such as status spongiosus, gliosis, and nerve cell loss. MRI can help to differentiate CJD from other dementing processes. PMID- 3045695 TI - [Combined radio-surgical treatment of tumors of the rectum and the recto-sigmoid junction. Review of the literature]. PMID- 3045696 TI - [Inguinal hernia: repair by extraperitoneal approach with an ivalon prosthesis. Our experience]. PMID- 3045697 TI - [Retroperitoneal leiomyosarcoma. Diagnostic and therapeutic role of surgical intervention by excision]. PMID- 3045698 TI - [Cystic pyeloureteritis. Presentation of a case and review of the literature]. PMID- 3045699 TI - Alkylation of striatal dopamine receptors abolishes stereotyped behavior but has no effect on dopamine stimulated adenylate cyclase activity. AB - The role of dopamine stimulated adenylate cyclase (AC) activity in behaviours elicited by apomorphine stimulation of striatal dopamine receptors was investigated. Rats were treated with the irreversible dopamine receptor alkylating agent N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) alone or after pretreatment with either a D1 or D2 receptor antagonist and subsequently challenged with apomorphine. Animals that received no antagonist pretreatment showed significantly decreased striatal concentrations of D1 and D2 receptors and an abolition of apomorphine induced sniffing behaviour despite showing no change in striatal AC activity. In the groups which received antagonist pretreatment the reduction in the sniffing response paralleled the reduction in D2 receptor concentration whereas the incidence of vacuous oral movements was inversely related. In no case were the behavioural responses associated with changes in AC activity. We conclude that these behavioural effects observed in response to dopamine stimulation by apomorphine may be mediated through another second messenger system. PMID- 3045701 TI - Tropane alkaloids. PMID- 3045700 TI - Prostaglandins, thromboxanes, leukotrienes, and related arachidonic acid metabolites. PMID- 3045702 TI - The biosynthesis of shikimate metabolites. PMID- 3045704 TI - Decreased energy expenditure as a cause of obesity in infants. PMID- 3045703 TI - The influence of alcohol on nutritional status. PMID- 3045705 TI - Taurine supplementation in cystic fibrosis. PMID- 3045706 TI - Journal of Medical Primatology Volume 3, 1974: An experimental model of alcohol feeding and liver injury in the baboon. By C.S. Lieber and L.M. DeCarli. PMID- 3045707 TI - 1,25-Dihydroxyvitamin D3 affects growth factors in bone cells. PMID- 3045709 TI - Evolution and critique of changes in the Jones criteria for the diagnosis of rheumatic fever. PMID- 3045708 TI - The immunology of rheumatic fever. PMID- 3045710 TI - Rheumatogenic streptococci reconsidered. PMID- 3045711 TI - Longitudinal study of poststreptococcal disease in Auckland; rheumatic fever, glomerulonephritis, epidemiology and M typing 1981-86. PMID- 3045712 TI - The rheumatogenic and nephritogenic strains of the group A streptococcus: the Kuwait experience. PMID- 3045713 TI - Rapid detection of group A streptococcal antigen for the clinician and the epidemiologist: accurate? cost-effective? useful? AB - Reviewing the literature describing rapid antigen detection for the diagnosis of group A streptococcal upper respiratory tract infections, one can conclude that the rapid aspect of these tests is clearly advantageous. To be able to have an answer in 5-20 minutes has distinct advantages with regard to therapy for patients. The specificity of most of these tests is acceptable for group A streptococci, although one must recognise that group C and group G streptococci can cause upper respiratory tract infection, and would not be detected by any group A antigen test. A remaining problem appears to be that of sensitivity. Although the range of sensitivity with various techniques and in the hands of different investigators may vary, it is difficult to ignore the fact that the sensitivity for many tests does not appear to be optimal. Furthermore, appropriate technical skill is required before these tests can be recommended for use in the hands of inadequately trained individuals, either in an office laboratory or in peripheral health care stations. There is also the concern that the cost for these diagnostic tests, unless purchased in large quantities, generally is greater than for the throat culture. This could have significant implications in some health care situations. One potential alternative for the office practitioner to consider is that if a rapid antigen test is negative, he or she should also perform a throat culture. This is the routine carried out in many practising clinicians' offices.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3045714 TI - Rheumatic fever in the United States: no longer a disease of the past. PMID- 3045715 TI - Establishment of a rheumatic heart disease registry in Taipei: an early appraisal. PMID- 3045716 TI - Mammographic screening for breast cancer: prospects for New Zealand. AB - The Cancer Society and the Department of Health invited a working group to make recommendations on screening by mammography. Mammography offers the best opportunity for preventing deaths from breast cancer. Randomised trials suggest that mortality can be reduced by about 30% in women over 50; the value of routine mammography in younger women is still uncertain. Apart from financial costs, the main drawback of mammography is that many women receive unnecessary investigation because of false-positive results. Careful design and monitoring of programmes is essential to ensure that the benefits of screening outweigh the disadvantages. In New Zealand there is a shortage of radiologists, pathologists, and clinicians who are skilled in the specialised techniques required for the screening of asymptomatic women. Decisions about routine screening should be delayed until pilot programmes have been established, with assessment of their effectiveness, economic efficiency, and social acceptability. Recommendations for the design of such programmes are made. PMID- 3045717 TI - Comparative study of norfloxacin and trimethoprim for the treatment of elderly patients with urinary tract infection. AB - One hundred elderly hospitalised patients, aged 70 to 97 years (mean 81.7 years; SD 7.1 years) with a urinary tract infection were entered into a randomised study to compare the efficacy, safety and tolerance of norfloxacin 400 mg twice daily for three days with trimethoprim 300 mg once daily for three days. Forty-two of 49 patients (86%) were cured with norfloxacin, compared with 35 of 51 (69%) with trimethoprim (p less than 0.05). No patient reported any side effects during treatment. Two patients treated with norfloxacin and three treated with trimethoprim developed a disturbance of liver function. Three deaths occurred within 28 days of treatment with trimethoprim but were unrelated to the treatment. In this study norfloxacin proved superior to trimethoprim for the treatment of uncomplicated urinary tract infections in elderly hospitalised patients. PMID- 3045718 TI - Single dose gentamicin treatment of urinary infections in children. AB - Sixty-nine children with urinary tract infections were randomly allocated to single dose gentamicin therapy (n = 39) or a seven day course of an appropriate antibiotic (n = 30). During the following six weeks the response to treatments did not differ and this was not altered by the child's clinical diagnosis, past history of infection or presence of radiological abnormalities. The poorest response was in those children with a history of recurrent infections (p less than 0.01) and/or radiological abnormality (p less than 0.02). Single dose therapy had significantly less suppression upon rectal (p less than 0.001) and periurethral (p less than 0.02) flora. There was a tendency for those not cured by single dose treatment to relapse whereas those treated by conventional therapy tended to be reinfected. PMID- 3045719 TI - The inspector-general of hospitals and the chief health officer. PMID- 3045720 TI - A New Zealand death from malaria. PMID- 3045721 TI - The engymetric determination of acute functional impairment in kidney graft function caused by cyclosporine A. AB - Engymetry offers a new means of continuously measuring nuclear radiation fields without any need of restricting the patient's mobility. In this study, kidney function was measured simultaneously and continuously for 6.6 h with engymetry, after application of a 12 h therapy dose of Cyclosporine A (CsA) of 4.0 +/- 1.8 mg per kg bodyweight (bw). 15 kidney transplanted patients participated in this study. 370 MBq 99Tcm-DTPA and 10 MBq 131I-OIH were injected during routine transplant scintigraphy. Renal function was monitored from the external disappearance curves of the tracers recorded by portable double radionuclide detectors. Renal impairment could be seen in the rising phase of CsA or coincided with the CsA maximum in 13 of the 15 patients. Under the impairment the half-life of 99Tcm-DTPA increased from 4.1 +/- 1.2 to 15.9 +/- 12.3 h (p less than 0.005) for 61 +/- 45 min and the half-life of 131I-OIH increased from 3.2 +/- 0.7 to 12.7 +/- 8.4 h (p less than 0.001) for 71 +/- 37 min. With noninvasive engymetry it is possible to detect renal functional impairment during long observation periods. Acute restriction in glomerular filtration and renal blood flow was discovered in human kidney graft recipients after the application of a low therapeutic CsA dose. PMID- 3045722 TI - The plasma clearance rate of 131I-insulin in diabetic and obese patients and healthy persons. AB - The plasma clearance rate of 131I-insulin was studied in diabetic and obese patients and healthy persons. The aim of the investigation was to find out whether an accelerated degradation of insulin-probably due to the metabolism of insulin by adipocytes-causes slight forms of the diabetic syndrome. For the evaluation of the data a three-compartment model was used whereby the third compartment reflects the degradation of the intravenously administered 131I insulin in the tissue. In this compartment 131I-insulin has a half life of 38.7 +/- 7.0 min for control persons (n = 19), 47.4 +/- 27.4 min for patients with a latent diabetes mellitus (n = 7), 34.2 +/- 8.5 min for the group with the subclinical diabetes (n = 5) and 47.6 +/- 16.2 min for the patients with an onset diabetes mellitus (n = 5). No significant difference was observed in the plasma clearance rate between healthy and diabetic persons. There is also no detectable correlation between the grade of obesity and the insulin clearance rate. PMID- 3045723 TI - Mediastinal Hodgkin's disease--a retrospective analysis. AB - Thirty-seven patients with mediastinal Hodgkin's disease treated from 1972 to 1982 at the Division of Oncology of Basel University Hospital were analyzed retrospectively for their characteristics and the influence of various prognostic factors on survival. Deceased patients were older (35 vs. 25 years) and more often had systemic symptoms (8/22 vs. 3/15). 44% of the patients presented with mediastinal bulk disease, 66% of whom are in remission. No statistically significant influence of sex and stage was evident on the outcome. Patients treated with multimodality therapy had a significantly better survival than patients treated with radio- or chemotherapy only. No significant difference was found in survival in comparison to the population without mediastinal involvement. PMID- 3045724 TI - Phase-II study with the combination of cisplatin and doxorubicin in advanced malignant mesothelioma of the pleura. AB - The effectiveness of combination chemotherapy with doxorubicin and cisplatin was studied in patients with advanced malignant pleuramesothelioma. 19 patients were treated intravenously with cisplatin 60 mg/m2/day on days 1 and 2, and with adriamycin 40 mg/m2 on day 3 every 4 weeks. 8/19 patients (46%) responded to chemotherapy; 2 achieved complete remissions (CR), and 6 went into partial remissions (PR). Median duration of response was 222 days. Median survival time of all patients was 370 days compared to a median survival of 273 days in a historical control group consisting of 30 patients treated by surgery only. Substantial toxicity was observed, mainly gastrointestinal. We conclude that the combination of cisplatin and adriamycin is effective in malignant pleuramesothelioma. However, the duration of treatment is limited by gastrointestinal toxicity. PMID- 3045725 TI - [Bromocriptine in chemotherapy-resistant, metastatic breast cancer. Results of the GO-MC-BROMO 2/82 AIO Study]. AB - In a case report a patient with metastatic breast cancer, who had chemotherapy resistance and hyperprolactinemia, showed a tumor remission following suppression of elevated prolactin levels with bromocriptine. Based on this observation, 18 patients with progressive metastatic breast cancer, chemotherapy resistance and hyperprolactinemia were treated with 10 mg bromocriptine/day in a prospective study. This treatment was in addition to chemotherapy, which was continued for 8 weeks in spite of progressive disease. In all patients, elevated prolactin levels (arithmetic mean 1388 +/- 201 mU/l before treatment) were suppressed to values under 100 mU/l. A partial remission was observed in one single patient which was not clearly attributed to suppression of plasma prolactin levels, but to delayed tumor remission. Side effects, mainly gastrointestinal disorders, were observed in all patients during therapy. In 6/18 patients treatment had to be stopped before end of study due to intolerable nausea and vomiting. It is concluded that suppression of elevated prolactin levels in progressive metastatic breast cancer patients is not effective in restoring tumor sensitivity to chemotherapy. PMID- 3045726 TI - Randomized trial of tamoxifen versus aminoglutethimide and versus combined tamoxifen and aminoglutethimide in advanced postmenopausal breast cancer. AB - In a randomized trial, 105 postmenopausal women with advanced carcinoma of the breast received tamoxifen or aminoglutethimide or combined tamoxifen and aminoglutethimide. No differences were found in the rate of responses and duration of responses between the treatment groups. Toxicity was significantly greater (p less than 0.01) in patients who received aminoglutethimide. PMID- 3045727 TI - Clinical interest and problems related to the measurement of the blind spot and the pericoecal region by means of programs SAPRO, SAPPAR, and BSPOT. AB - Difficulties encountered when measuring the differential light sensitivity of the blind spot and its surroundings and the clinical interest of such measurements are reported. The determination of the differential light sensitivity across step gradients, as well as abrupt changes observed at and close to the blind spot, may pose severe problems, because both are strongly influenced by even slight fixation instability. However, other factors reported herein may be equally important. The insight gained here may be cautiously extrapolated to the difficulties (and to the efforts to overcome them) encountered when measuring other scotomata in the visual field having sensitivity distributions similar to those of the blind spot. The sensitivity distribution in the blind spot and the pericoecal region may be important in a number of disease entities, particularly in glaucoma and neurologic diseases. PMID- 3045728 TI - Management of the blepharoplasty patient with ptosis. AB - Patients who request correction of both dermatochalasis and acquired blepharoptosis require separate evaluation of each condition. The blepharoplasty is done in the standard way, depending on the amount of skin redundancy and fat herniation. The method of ptosis repair is dependent on whether Muller's muscle or levator aponeurosis is weak, the degree of ptosis and levator function, and the position of the upper eyelid crease. A tarsal Muller's muscle resection is employed in cases where the ptosis is minimal, the upper eyelid crease is normal, or the phenylephrine test is positive. A levator advancement is done if the ptosis is due to a levator aponeurosis dehiscence or attenuation with a higher than normal upper eyelid crease and thinning of the eyelid. Both procedures are easily combined with an upper eyelid blepharoplasty. PMID- 3045730 TI - The Edwin Smith Surgical Papyrus: an analysis of the first case reports of spinal cord injuries. PMID- 3045731 TI - The contributions of plastic surgery to care of the spinal cord injured patient. AB - Plastic surgeons have contributed to the understanding of pressure sore pathophysiology and prophylaxis. Increasingly sophisticated surgical techniques such as myocutaneous or innervated flaps add to the reliability and durability of repairs. The majority of quadriplegics may benefit from surgical restoration of active elbow extension, lateral pinch and grasp. Prolonged postoperative care in bed or immobilisation of the upper limb demands that patients should understand fully all that the reconstructive procedure involves. The nature and importance of subsequent rehabilitation must be appreciated by the patient so that he will be motivated to achieve the best possible result. Functional electrical stimulation may find an increasing role in the years to come. PMID- 3045729 TI - Human cDNAs rap1 and rap2 homologous to the Drosophila gene Dras3 encode proteins closely related to ras in the 'effector' region. AB - We have characterized two new ras-related genes rap1 and rap2 from a human cDNA library, by hybridization with the Drosophila Dras3 gene at low stringency conditions. The rap1 and rap2 genes encode proteins of 184 and 183 amino acid respectively with molecular weights of 20.9 kd and 20.7 kd. These proteins are 53% and 46% identical to the human K-ras protein and share several properties with the classical ras proteins. The C-terminal cysteine involved in the membrane anchoring as well as the GTP binding regions of the p21 ras proteins are present in the rap proteins suggesting that these proteins could bind GTP/GDP and have a membrane localization. The most striking difference between the rap and ras proteins resides in their 61st amino acid. As in the Drosophila Dras3 protein, both rap proteins have a threonine instead of the glutamine found at position 61 of the classical ras proteins. Furthermore the putative effector domain of the ras proteins is strictly conserved in the rap1 protein whereas only one amino acid difference is found in the rap2 protein. This suggests that the rap proteins might interact with the same effector as the ras proteins. PMID- 3045732 TI - Effects of pin care on pin reactions in adults with extremity fracture treated with skeletal traction and external fixation. PMID- 3045733 TI - Computed tomography of rheumatologic disorders. Part I. AB - The advent of computed tomographic (CT) scanning has initiated a revolutionary approach to the evaluation of articular disease. CT affords the opportunity to noninvasively evaluate both osseous and soft tissue structures during a single diagnostic examination. An optimal CT study demands proper technique, including a localization image, appropriate slice thickness and spacing, and window/level manipulation. The examination must be tailored to the specific indication for the study, which may include degenerative processes, inflammatory disease, or unexplained clinical symptomatology. PMID- 3045734 TI - Patellar metastases. A case report and review of the literature. AB - A case of metastasis to the patella from adenocarcinoma of the lung is reported. The clinical presentation was similar to septic arthritis of the knee. Histopathologic examination established the metastatic nature of the lesion; this is the first case in which a patellar lesion was the only evidence of distant metastasis. Metastatic tumors of the patella are rare. Three cases of metastasis to the patella have been described from squamous cell carcinoma of the lung in the English literature. We report the first case of patellar metastasis from adenocarcinoma of the lung. PMID- 3045736 TI - [Methodological problems in determining the time of clinical death]. PMID- 3045735 TI - [Hemodynamics and myocardial blood flow after orthotopic transplantation of a donor heart in the early postoperative period]. PMID- 3045737 TI - [A method for cloning hemopoietic precursor cells in the bone marrow microenvironment]. PMID- 3045738 TI - [A method for the comparative ultra-acoustic study of the membranes of human hollow organs in different forms of pathology]. PMID- 3045739 TI - [Current aspects of the problem of classifying traumatic shock in the clinic]. PMID- 3045740 TI - Autoradiographic determination of regional cerebral blood flow during hypoglycemia in newborn dogs. AB - To ascertain the regional cerebral blood flow (CBF) responses to hypoglycemia, nine newborn dogs were treated with insulin to blood glucose concentrations ranging from 1 to 35 mg/dl (mean 22 mg/dl). Systemic physiologic monitoring revealed no differences in mean arterial blood pressure, heart rate, paO2, paCO2, pHa, or blood lactate in the hypoglycemis animals and five normoglycemic controls. Significant increases in CBF occurred in 17 of 20 analyzed structures of brain in the hypoglycemic puppies, ranging from 158 to 446% of the normoglyycemic values. The percent increases in CBF were greatest in brainstem structures compared to other major regions of brain. A positive correlation existed between mean arterial blood pressure and cerebral cortical blood flow, suggesting a loss of CBF autoregulation during hypoglycemia. The pathophysiologic mechanism for the elevations in regional CBF might relate to stimulation of beta adrenergic receptors in brain, as has been shown in adults. PMID- 3045741 TI - Indomethacin restricts cerebral blood flow during pressure ventilation of newborn pigs. AB - Unanesthetized newborn pigs were studied to evaluate the immediate (10 min) and delayed (45 min) effects of increased ventilation pressure coupled with cyclooxygenase inhibition. Cardiac output and cerebral blood flow were measured at a low (5 cm H2O) and high (30 cm H2O) mean airway pressure (Paw) before and 45 min after 5 mg/kg of indomethacin. In a second group, these parameters were also measured 10 min after indomethacin was given during ventilation at a Paw of 30 cm H2O. Before treatment with indomethacin, increasing Paw decreased cardiac output without affecting cerebral blood flow. Baseline (Paw = 5 cm H2O) cerebral blood flow decreased 40% 45 min after indomethacin treatment. Adding the stress of a ventilation-induced drop in cardiac output did not further depress cerebral blood flow. When indomethacin was administered during high Paw, cerebral blood flow decreased markedly within 10 min. Cerebral oxygen consumption was maintained by increasing oxygen extraction. Therefore, indomethacin decreases cerebral blood flow at a high Paw. The fall in cerebral blood flow decreases brain oxygen delivery. However, cerebral oxygen consumption is maintained by an increase in oxygen extraction. PMID- 3045742 TI - Plasma vasopressin, renin, and catecholamines during nitroprusside-induced maternal and fetal hypotension in sheep. AB - The release of vasopressin, renin, and catecholamines by the fetus during either maternal or fetal hypotension was examined in chronically catheterized fetal lambs. Nitroprusside was infused intravenously for 1 h into seven pregnant ewes (maternal hypotension) or nine fetal lambs (fetal hypotension); the rates were adjusted to achieve a 15 to 30% decrease in mean blood pressure. During maternal hypotension, mean +/- SE vasopressin in maternal plasma increased from 1.2 +/- 0.2 pg.ml-1 to 208 +/- 153 pg.ml-1 and plasma renin activity increased from 1.5 +/- 0.3 ng.ml-1.h-1 to 6.6 +/- 1.6 ng.ml-1.h-1. Fetal vasopressin and plasma renin activity also increased during the same interval from 1.1 +/- 0.3 to 16.9 +/- 7.5 pg.ml-1 and 3.7 +/- 1.1 to 10.5 +/- 2.85 ng.ml-1.h-1, respectively; but no changes were observed in fetal blood pressure, heart rate, or acid base status. During fetal hypotension, mean vasopressin in fetal plasma increased from 4.3 +/- 3.4 pg.ml-1 to 1054 +/- 772 pg.ml-1, plasma renin activity increased from 5.7 +/- 2.2 ng.ml-1 to 22.2 +/- 7.1 ng.ml-1.h-1, and total catecholamines from 174 +/- 58 pg.ml-1 to 810 +/- 416 pg.ml-1. There was no change in fetal heart rate, acid base status, osmolality, or sodium concentration. The fetus became and remained hypertensive for at least 1 h after the end of infusion. This prolonged hypertension was associated with elevated levels of vasopressin and plasma renin activity. Peak vasopressin levels were proportional to the total nitroprusside dose in both the ewe and fetus (maternal r = 0.796, fetus r = 0.870).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3045743 TI - [School health services. III. The 1970's--trends toward the integration of school health services and pediatrics]. PMID- 3045744 TI - The effect of prostacyclin and its analogues on cysteamine-induced duodenal ulceration of rats. AB - The effect of prostacyclin and its analogues--6-keto-prostaglandin-F1 alpha and 6 beta-prostaglandin-I1--on cysteamine-induced duodenal ulceration of rats was investigated. It seems that neither prostacyclin, nor its analogues have anti ulcerogenic effect in this model. PMID- 3045746 TI - Cimetidine and placebo-cimetidine in the treatment of patients with chronic gastric ulcer (a multiclinical randomized double-blind comparative study). AB - The efficacy of a 4-week cimetidine treatment was examined by a double-blind randomized study in 37 outpatients with endoscopically verified chronic gastric ulcer. The patients received a daily dose of 3 times 1 tablet and, at night before going to bed 2 more tablet, thus a total amount of 1 g cimetidine, or cimetidine-placebo, but in case of complaints they could take in addition a mixed alkaline powder. Patients not recovering in response to a 4-week treatment, were then administered daily 5 tablets of cimetidine up to their complete recovery. Endoscopic, laboratory and clinical examinations were carried out every other week. As a result of a 4-week treatment, 56% of the cimetidine group recovered. The difference was not significant (P less than 0.2). The size of the ulcer and the intensity of the complaints were reduced significantly in both groups. The decrease in the size of the ulcer was significantly greater in the first two weeks of cimetidine treatment than in the cimetidine-placebo group (P less than 0.05). This favourable dynamics of ulcer healing was not felt in the second two weeks of treatment, and after four weeks there was no difference in the size of the residual ulcer to between the two groups. Cimetidine seemed to be a suitable drug for treating chronic gastric ulcer, since its healing rate proved to be better than that of placebo, the gain in weight also was favourable and there were no side-effects. PMID- 3045745 TI - Function of monocytes in patients with systemic sclerosis. AB - Functions of monocytes from the peripheral blood of 23 patients with systemic sclerosis were investigated in vitro. The yeast phagocytosis, opsonized yeast phagocytosis and binding of EA (erythrocyte-antibody) particles were found to be normal. A depressed chemotactic response was demonstrated against a zymosan activated, complement-derived chemotactic factor. In 12 cases, monocytes were cultured for 168 hours. By the 5th and 7th days, the initially depressed chemotactic activity of monocytes returned to normal as compared to controls. This fact supports the speculation that the decreased chemotaxis cannot be caused by an intrinsic abnormality of monocytes/macrophages in systemic sclerosis. PMID- 3045747 TI - [The role of radiobiology in radiotherapy]. PMID- 3045748 TI - [Evaluation of the diagnostic efficacy of ultrasound examination combined with body X-ray computed tomography in pancreatic diseases]. PMID- 3045749 TI - Educational outcomes. Review of the literature. PMID- 3045750 TI - [Coping crown construction]. PMID- 3045751 TI - [Viking-age skeleton exhibiting pathologic changes in the dentition and maxillary and mandibular bones--scurvy or leprosy?]. PMID- 3045752 TI - [Examination of teeth and tooth-fragments from three burial sites in the White Nile Valley of southern Sudan]. PMID- 3045753 TI - Molecular consequences of truncations of the first exon for in vitro splicing of yeast actin pre-mRNA. AB - A defined minimum length of the first exon is required for the generation of spliced products from a synthetic yeast actin mRNA-precursor in vitro. If the first exon is 1, 2, 3 or 5 nucleotides long, only the first step of the splicing reaction can take place. A transcript starting with the first nucleotide of the intron does not get converted into any of the normally obtained splicing products or intermediates. On the other hand, spliceosome assembly does not depend on the presence of a first exon. PMID- 3045754 TI - Transcription initiation at the tet promoter and effect of mutations. AB - We have identified the startpoint for transcription in vitro of the tetracycline resistance gene (tet) of pBR322 and several deletion and insertion mutations which alter tet promoter structure. Tetracycline resistance in host bacteria correlates qualitatively with the efficiency of DNA fragments from these plasmids to promote tet transcription in vitro. Only in active promoters could we find by computer analysis promoter structures in which the -10 and -35 sequences and the relative spacing of the two regions agree with consensus sequence determinants. These data support the current model of the E. coli promoter sequence. Two promoter mutants gave heterogeneous 5' termini with additional A residues not encoded by the DNA sequence. PMID- 3045755 TI - Identification of two factors which bind to the upstream sequences of a number of nuclear genes coding for mitochondrial proteins and to genetic elements important for cell division in yeast. AB - Two abundant factors, GFI and GFII which interact with the 5' flanking regions of nuclear genes coding for proteins of the mitochondrial respiratory chain have been identified. In one case (subunit VIII of QH2: cytochrome c oxidoreductase) the binding sites for both factors overlap completely and their binding is mutually exclusive. For the other 5' regions tested the GFI and GFII binding sites do not coincide. Interestingly, binding sites for GFI and GFII are also present in or at the 3' ends of the coding regions of two genes of the PHO gene family and in DNA elements important for optimal ARS and CEN function respectively. The sites recognized by GFI conform to the consensus RTCRNNNNNNACGNR, while those recognized by GFII contain the element RTCACGTG. We speculate that GFI and GFII may play a role in different cellular processes, dependent on the context of their binding sites and that one of these processes may be the coordination of the expression of genes involved in mitochondrial biogenesis with the progress of the cell cycle. PMID- 3045756 TI - A general method of in vitro preparation and specific mutagenesis of DNA fragments: study of protein and DNA interactions. AB - Specific, end-labeled DNA fragments can be simply and rapidly prepared using the polymerase chain reaction (PCR). Such fragments are suitable for use in DNase I protection footprint assays, chemical sequencing reactions, and for the production and analysis of paused RNA polymerase transcription complexes. Moreover, a general means of introducing a specific mutation at any position along the length of such PCR-generated fragments is described. These procedures, which can circumvent the need for large-scale phage or plasmid growths, preparative gel-electrophoresis and the screening of molecular clones, can facilitate the rapid study of sequence-specific interactions of proteins and DNA. A rapid means of removing excess oligonucleotide primers from completed PCRs is also described. PMID- 3045758 TI - Kinetic analysis of an E.coli phenylalanine-tRNA synthetase mutant. AB - A mutation in the pheS gene, encoding phenylalanyl-tRNA synthetase, in E. coli NP37 confers temperature-sensitivity on the organism. A five-fold increase in tRNA(phe) levels complements the mutation. Analysis of the kinetic properties of the mutant enzyme indicates that the KM is 20-fold higher than the wild-type and the dissociation constant of the tRNA(phe)-synthetase complex for the mutant is at least 10-fold higher. These results indicate that the mutation in E. coli NP37 directly affects the tRNA(phe) binding site on the cognate synthetase. PMID- 3045757 TI - The 9S RNA precursor of Escherichia coli 5S RNA has three structural domains: implications for processing. AB - The secondary structure of the 9S RNA precursor to ribosomal 5S RNA in Escherichia coli has been determined using chemical reagents and ribonucleases in combination with a reverse transcription procedure. The 9S RNA precursor was generated in vitro by T7 RNA polymerase, and the rrnB operon terminator, T1, was able to terminate the in vitro transcript. The secondary structure model exhibits three structural domains corresponding to a 5' region, a mature region and a terminator region. The mature domain is structurally identical to 5S RNA, and the ribosomal proteins L18 and L25 are able to bind to the precursor. The processing endoribonuclease RNase E cleaves between the structural domains. Moreover, an intramolecular refolding of the nascent transcript must take place if the current view of RNase III processing stems is correct. PMID- 3045759 TI - A novel yeast secretion signal isolated from 28K killer precursor protein encoded on the linear DNA plasmid pGKL1. AB - Saccharomyces cerevisiae harboring linear dsDNA plasmids, pGKL1 and pGKL2, secretes a killer toxin consisting of 97, 31 and 28 kilodalton subunits (Nucleic Acids Res., 15, 1031-1046, 1987). We isolated the DNA encoding the N-terminal pre sequence of the 28K precursor protein and constructed a new secretion vector in S. cerevisiae. Mouse alpha-amylase fused to the 28K signal sequence was secreted into the culture medium with a high efficiency similar to those fused to the mating factor alpha and 97K-31K killer signal sequences. This data clearly indicates that 28K presequence functions as a secretion signal. Glycosylated and nonglycosylated alpha-amylase molecules were detected in the culture medium. The secretion of alpha-amylase was blocked by sec18-1 mutation. The secreted alpha amylase recovered from the medium was found to migrate faster in SDS polyacrylamide gel than the precursor form of alpha-amylase synthesized in vitro. These lines of evidence suggest that mouse alpha-amylase fused to 28K killer signal sequence was processed, glycosylated and secreted through the normal secretion pathway of the yeast. PMID- 3045760 TI - Sequence analysis and transcriptional regulation of the Escherichia coli grpE gene, encoding a heat shock protein. AB - We have sequenced the Escherichia coli grpE gene and shown that it encodes a 197 amino acid residue protein of 21,668-Mr. The predicted N-terminal amino acid sequence, as well as the overall amino acid composition agree well with that of the purified protein. From Northern analysis, we have shown that transcription of the grpE gene is under heat shock regulation, i.e., there is a rapid and transient increase in the rate of synthesis of grpE mRNA upon a shift-up in temperature. Forty-six bases upstream of the structural gene is a sequence closely related to the consensus heat shock promoter identified by Cowing et al. [Proc. Natl. Acad. Sci. U.S.A, 82, 2679-2683]. We have shown by S1 mapping and RNA sequencing that this is indeed the promoter for the grpE mRNA. It appears that all discernable transcription initiates only from this promoter, even under non-heat shock conditions. PMID- 3045761 TI - Defining the consensus sequences of E.coli promoter elements by random selection. AB - The consensus sequence of E.coli promoter elements was determined by the method of random selection. A large collection of hybrid molecules was produced in which random-sequence oligonucleotides were cloned in place of a wild-type promoter element, and functional -10 and -35 E.coli promoter elements were obtained by a genetic selection involving the expression of a structural gene. The DNA sequences and relative levels of function for -10 and -35 elements were determined. The consensus sequences determined by this approach are very similar to those determined by comparing DNA sequences of naturally occurring E.coli promoters. However, no strong correlation is observed between similarity to the consensus and relative level of function. The results are considered in terms of E.coli promoter function and of the general applicability of the random selection method. PMID- 3045762 TI - The role of the excision-repair enzymes in mutation-induction by cis-Pt(NH3)2Cl2. AB - Mutation induction by cis-Pt(NH3)2Cl2 (cisplatin) has been shown to be absent in E.coli strains carrying a deletion of the uvrB gene (1). This suggested that excision-repair, which is normally thought to be error-free, is involved in mutation induction with cisplatin. Here, the role of the excision repair enzymes UvrA, UvrB and UvrC is investigated using E.coli strains with different repair capacities. It is shown that cisplatin induced mutagenesis is dependent both on UvrA and UvrB but not on UvrC. Of the UvrB enzyme the N-terminal 113 aminoacids are sufficient for mutation induction by cisplatin. PMID- 3045764 TI - Nucleotide sequence of the glpR gene encoding the repressor for the glycerol-3 phosphate regulon of Escherichia coli K12. PMID- 3045763 TI - A plastid fragment from the psbE-psbF coding region in the mitochondrial genome of Oenothera berteriana. PMID- 3045765 TI - pYAC-RC, a yeast artificial chromosome vector for cloning DNA cut with infrequently cutting restriction endonucleases. PMID- 3045766 TI - A look back; a look ahead. PMID- 3045768 TI - [Clinical value of enzymatic studies in lung cancer]. PMID- 3045767 TI - Public health nursing: a photographic study. PMID- 3045769 TI - [Dr. Stanislaw Ignacy Frenkel (29 November, 1917-2 February 1987]. PMID- 3045770 TI - [Results of the treatment of non-lymphoblastic leukemia with small doses of cytosine arabinoside]. PMID- 3045771 TI - [Capacity of selected bacteria to degrade elastin and their effect on the elastase-antielastase balance in studies in vitro]. PMID- 3045772 TI - [Barotrauma as a complication of controlled respiration]. PMID- 3045774 TI - [Clinical evaluation of beclomethasone dipropionate aerosol in patients with bronchial asthma]. PMID- 3045773 TI - [Comparison of bronchodilating effectiveness of oxitropium and fenoterol aerosols in bronchospastic conditions]. PMID- 3045775 TI - [Acute adult respiratory distress syndrome (ARDS): progress in studies on its pathogenesis, diagnosis and treatment]. PMID- 3045776 TI - [The role of lysosomal enzymes in lung damage in adult respiratory distress syndrome (ARDS)]. PMID- 3045778 TI - [Effectiveness of fendiline in ischemic heart disease]. PMID- 3045777 TI - [Current views on non-invasive treatment of hemorrhaging esophageal varices]. PMID- 3045779 TI - [New aspects of the treatment of cardiogenic shock]. PMID- 3045780 TI - Effects of diaper types on diaper dermatitis associated with diarrhea and antibiotic use in children in day-care centers. AB - Infants and toddlers in day-care centers have a relatively high frequency of diarrhea and/or oral antibiotic use, and may be at increased risk of developing diaper dermatitis when diapered. A six-month, prospective, double-blind study was conducted in day-care centers in Houston, Texas, to determine the frequency of diarrhea, antibiotic use, and diaper dermatitis in infants and toddlers wearing conventional (cellulose-only core) disposable diapers or disposable diapers with a core of absorbent gelling material (AGM) and cellulose. A questionnaire was administered weekly to the day-care staff to gather health information, and weekly visual examinations were made of children for diaper dermatitis. The frequency of diarrhea was 1.9 episodes per child-year and that of antibiotic use was 3.3 courses per child-year. Infants diapered in disposable diapers with AGM had a significantly (P 0.032) lower mean grade of diaper dermatitis during diarrhea episodes and a lower (P 0.054) mean grade during antibiotic use, compared to those diapered in conventional disposable diapers. There was no significant difference between groups with regard to isolation of Staphylococcus aureus or Candida albicans from superficial skin cultures of the diapered area. The results indicate that diarrhea and antibiotic use occur frequently in children in day-care centers, and that the severity of diaper dermatitis is less in children wearing AGM disposable diapers than those wearing conventional disposable diapers in that setting. PMID- 3045783 TI - Findings in child deaths registered as sudden infant death syndrome (SIDS) in Madison, Wisconsin. AB - A review of a sequential series has been carried out on 60 Madison infants whose deaths had been registered as SIDS. In 18% it was possible to identify lesions that could have given a diagnosis other than SIDS on these same infants. When the cases were assessed on a multifactorial basis, 67% of the infants had a variety of lesions present, which in combination would probably have made their deaths explicable. It was only in children dying before the age of 18 weeks that more than half of the deaths were not explicable. The generally accepted concept of SIDS as a completely unexplained death would appear to apply only if the term is confined to a single aspect of pathology--the terminal anatomic state. If the total pathologic situation of the child is taken into consideration, the diagnosis of a completely unexplained death applies to only about a third of the cases examined in Madison. The current autopsy approach to sudden and unexpected deaths from a causal viewpoint requires critical reappraisal. PMID- 3045782 TI - Neonatal ovarian torsion: report of three cases and review of the literature. AB - Ovarian cysts are common incidental findings in term infants and, if unusually large, may result in dystocia, torsion, or rupture. Torsion and infarction of a normal ovary tend to occur in older childhood. During a 4-month period, 3 cases of neonatal ovarian torsion were observed after antenatal ultrasonography had detected fetal pelvico-abdominal cystic lesions. The three infants were explored between 4 and 16 days of age. Ovarian torsion was right-sided in all 3, and 1 ovary had been autoamputated. The resected specimens were nontense, thin-walled cysts, filled with hemorrhagic fluid, that measured between 4.5 and 8 cm in diameter. Microscopically, focal calcification and widespread necrosis precluded recognition of underlying histologic landmarks. Neonatal ovarian cysts or cystic ovaries greater than 4 cm in diameter should be excised, even if asymptomatic, because they are prone to, or have undergone, torsion. PMID- 3045781 TI - Impetigo contagiosa: a comparison of erythromycin and dicloxacillin therapy. AB - One hundred patients with impetigo were prospectively enrolled in a study to determine the current etiology and comparative therapeutic efficacy of two oral antimicrobial agents active against both group A beta-hemolytic streptococci (GABS) and Staphylococcus aureus. After obtaining a bacterial culture from a representative impetiginous lesion, the children were randomized to receive 10 days of either erythromycin (40 mg/kg/day) or dicloxacillin (25 mg/kg/day). S. aureus alone was isolated from 46 children, and in association with GABS from 25 children. GABS alone was isolated from nine patients. Of the 59 evaluable children with S. aureus isolates, 28 of 29 treated with erythromycin and 29 of 30 treated with dicloxacillin were cured or improved on follow-up examination. Thus, we conclude that erythromycin is the drug of choice for impetigo in our midwestern locale because of its high efficacy and relatively low cost. PMID- 3045785 TI - Office evaluation of dementia. How to arrive at a clear diagnosis and choose appropriate therapy. AB - About half of patients presenting with dementia have Alzheimer's disease, which cannot be prevented or reversed. Treatment of these patients is aimed at improving their quality of life and helping their families provide supportive care. About 15% of patients with dementia can be successfully treated. A detailed patient history, thorough physical examination, and appropriate laboratory testing are important to identify the treatable causes of dementia, such as normal-pressure hydrocephalus, Korsakoff's psychosis, and drug toxicity. PMID- 3045784 TI - Fetus in fetu: a fetiform teratoma. AB - Examination of a retroperitoneal fetus in fetu, diagnosed preoperatively, revealed previously unreported histologic findings in the vascular pedicle and membranous capsule that indicated that these structures are not "umbilical cord" or "amnion." These findings include nervous innervation of both structures and a well-defined lamina elastica interna and vaso vasorum in the artery of the vascular pedicle. Thus, strong support is provided for the theory that many examples of fetus in fetu are remarkably complex, well differentiated, highly organized teratomas. Additional arguments that favor the identity of fetus in fetu and teratoma are presented. PMID- 3045786 TI - Urinary tract infections in elderly patients. PMID- 3045788 TI - Prolonged pregnancy. When to wait, when to intervene. AB - Careful antepartum testing and close observation of fetal well-being during labor and delivery are essential when pregnancy is prolonged. However, if the expected date of confinement is well documented and the pregnancy is at 43 weeks, a trial of labor or primary cesarean section is indicated because the risk of perinatal morbidity beyond 43 weeks is significantly increased. PMID- 3045787 TI - How to help patients cut down on saturated fat. A simple strategy. PMID- 3045789 TI - Genital herpes. How to relieve patients' physical and psychological symptoms. AB - Genital herpes simplex virus is being encountered at an increasing rate by the primary care physician. Recurrences of this disease create not only medical but psychological and social problems, of which the physician must be aware. Although acyclovir (Zovirax) has become a useful palliative tool, compassion, sensitivity, and understanding are essential in the treatment of this disease. Physician provided education is still currently the thrust for prevention. PMID- 3045790 TI - Preventing the spread of infectious disease. Precautionary measures for the office setting. PMID- 3045791 TI - Microsomal enzyme induction, lipoproteins and atherosclerosis. AB - The liver is the principal site for the synthesis and elimination of lipoproteins circulating in plasma, and alterations in hepatic function influence plasma lipoprotein levels. High HDL-C/T-C and apo A-I/apo B ratios, which are characteristic of a low coronary risk, have been typical of healthy subjects with high microsomal enzyme activity in the liver. Microsomal enzyme inducing drugs such as phenytoin, phenobarbital and carbamazepine, and also alcohol, influence serum lipid and apoprotein concentrations. The inducers increase the concentrations of hepatic microsomal enzyme and apo A-I mRNA, and also proteins and phospholipids. They similarly increase serum HDL-C and apo A-I levels and the HDL-C/LDL-C ratio, powerful protective factors against coronary heart disease. These parameters parallel hepatic protein and phospholipid concentrations, and microsomal enzyme activity as assessed by liver cytochrome P-450 or antipyrine kinetics. Serum LDL-C levels are inversely proportional to hepatic cytochrome P 450 concentrations. Experimental studies indicate that phenobarbital retards cholesterol accumulation in the arterial wall and the formation of atherosclerotic plaque. A decreased mortality rate from coronary heart disease has been reported for subjects who take enzyme inducers, drugs or alcohol, whereas impairment of hepatic microsomal function may promote atherogenesis. In addition to drugs non-pharmacological factors such as dietary constituents and physical exercise may influence hepatic microsomal function and hence improve the serum lipoprotein profile. These observations, which connect microsomal inducers, liver lipids and proteins, serum lipids and apoproteins characteristic of a low risk of atherosclerotic disease and low incidence of coronary deaths, lead to the conclusion that the activation of liver microsomal function can prevent atherogenesis in man. PMID- 3045792 TI - Actions and interactions of indomethacin and diazepam on performance in healthy volunteers. AB - Objective and subjective effects on performance of single oral doses of indomethacin (IM) 50 mg and 100 mg and diazepam (DZ) 10-15 mg, alone and in combination, were investigated in two double-blind studies conducted with parallel groups of healthy drug-naive student volunteers. Objective and subjective effects were measured at baseline as well as 0.5 and 1.5 hours after treatment. DZ significantly impaired performance in digit symbol substitution, letter cancellation, tracking and flicker fusion tests. It also induced exophoria and caused subjective (visual analogue scales) drowsiness, mental slowness, clumsiness and impaired overall performance. IM proved rather inactive when slightly impairing flicker fusion and digit substitution, and subjectively rendering the subjects clumsier. The combined effects of IM and DZ did not differ from those obtained with DZ alone. Both IM and DZ induced dizziness and their effects in this respect were additive when the drugs were used in combination. It is concluded that single therapeutic doses of indomethacin do not produce major psychomotor effects and do not in this respect increase the effects of diazepam. However, the feeling of dizziness, a side-effect common to both these drugs, may be additive when the drugs are used in combination. PMID- 3045794 TI - Treating depression in acute stage: biochemical and clinical aspects. AB - The treatment of depression has advanced dramatically over the last 50 years: electroconvulsion, monoamine oxidase inhibitors (MAOIs), cyclic antidepressants - in chronological order. The amine theories have been giving guidance in the development of new antidepressant drugs and have thus been important in the selectivity of drugs acting, for example, primarily on 5-hydroxytryptamine (5-HT) reuptake. Drugs like citalopram, fluoxetine, fluvoxamine, and paroxetine are very selective in their 5-HT reuptake inhibition, so that in clinical practice they can be tested for the specific importance of this system in depression. Research on cerebrospinal fluid (CSF) has for many years been restricted to amine metabolites, but we have now been able to measure the parent neurotransmitters, thanks to international cooperation. CSF adrenaline was low, noradrenaline normal, and serotonin and dopamine increased in untreated depression. This calls for major revision of the amine theories. In addition, our animal studies have shown marked increase in CSF adrenaline after MAOIs, and long-term effect of citalopram on CSF noradrenaline. This indicates the need for enlargement of the number of transmitters in the elucidation of the biology of depression and its treatment, and that interactive counterregulations may lessen the initial selectivity of antidepressant drugs. PMID- 3045793 TI - Analysis by immunofluorescence of alpha 1-antitrypsin levels on the surfaces of Trichomonas vaginalis strains differing in virulence. PMID- 3045795 TI - Pharmacological management of treatment-resistant depression. AB - Treatment-resistant depression might be considered as the failure of two families of antidepressant therapy given sequentially at sufficient dosage, for an adequate length of time and with continuous compliance. Patients with treatment resistant depression should be assessed as though they were new referrals, and factors which may be contributing should be dealt with whenever possible. A sequence of therapies based on a tricyclic antidepressant and a monoamine oxidase inhibitor, each combined with putative adjuvant therapies, is proposed as a working model pending further studies and developments. PMID- 3045796 TI - Receptor interactions of dopamine and serotonin antagonists: binding in vitro and in vivo and receptor regulation. AB - The advent of receptor binding techniques has provided new ways of studying the mechanism of action of drugs. In vitro radioligand binding is now currently applied to investigate the specificity or multiple action of compounds. By using the same technique, the binding affinity of a drug can be measured for a variety of neurotransmitter, drug, peptide and ion channel receptor binding sites, providing the drug's receptor binding profile (LEYSEN et al. 1981; LEYSEN 1984). However, in vitro receptor binding is only the initial step in the investigation of drug-receptor interactions. Investigations in vivo are required to allow evaluation of how and where a drug acts. In fact, the study of drug-receptor interactions comprises three main stages: (a) in vitro radioligand receptor binding; (b) in vivo receptor binding, providing information on the accessibility of the drugs to the receptors localized in various central and peripheral tissues, on the drug potency for occupying various receptors, on the duration of receptor occupation and on the relationship between the degree of receptor occupation and pharmacological effects; and (c) the study of receptor regulation: the effect of chronic drug treatment on receptor alterations compared with alterations in functional responses in vivo. In this article, we will illustrate the three stages of investigation of receptor interactions and discuss the relevance and importance of the findings, using as examples three drugs known in psychopharmacological research: (a) the neuroleptic haloperidol, a prototype of a dopamine D2 antagonist: (b) Setoperone, a potential antipsychotic agent with very potent serotonin S2 and moderate D2 antagonistic activity (CEULEMANS et al. 1985; LEYSEN et al. 1986); and (c) ritanserin, a potent and long-acting S2 antagonist (LEYSEN et al. 1985), which has revealed therapeutic activity in dysthymia and negative symptoms of schizophrenia (REYNTJENS et al. 1986; GELDERS et al. 1986). Particular attention will be paid to the problem of receptor regulation. We challenge the general applicability of the receptor regulation theory, which states that persistent receptor stimulation causes desensitisation and receptor downregulation, whereas chronic deprivation of receptor stimulation leads to supersensitivity and receptor upregulation. Recent research has revealed that the theory does not hold for S2 receptor alterations, which were found to downregulate following chronic receptor blockade. PMID- 3045797 TI - Is there a long-term protective effect of mood-altering agents in unipolar depressive disorder? AB - Major depression is common, often severe, and usually recurrent, and it carries an excess risk of mortality due to medical illness as well as suicide. At referral centers, recurrence risk averages 85% within 2-3 years of full recovery from an acute episode. The natural history of major depression is highly variable, but typically episodes las ca. 6 months, with cycles of ca. 2 years. Yet, most long-term treatment studies are limited to the year or two following recovery from an acute episode. Accordingly, available evidence best supports a relapse-preventing effect of tri-cyclic antidepressants or lithium within the first months after apparent recovery but is less compelling regarding prevention of later recurrences of new episodes. Other treatments have not been evaluated systematically. The hypothesis that bipolarlike, but apparently unipolar, patients might respond selectively to lithium maintenance requires further testing. Knowledge of long-term dose-benefit and dose-risk relationships is starting to emerge for lithium, but these relationships remain inadequately tested for antidepressants. Actual levels of clinical treatment of major depression appear to fall short of the ideal, and much additional research and education is required to improve care in this very common disorder. PMID- 3045798 TI - Basic and clinical aspects of the activity of the new monoamine oxidase inhibitors. AB - The clinical relevance of the neurobiochemical and pharmacological properties of the new generation of monoamine oxidase inhibitors (MAOIs) is reviewed and discussed. The most distinctive characteristics of these drugs are the selectivity, competitive nature and reversibility of their MAO-inhibiting action. The most selective MAO-A inhibitors are moclobemide and brofaromine, with ratios between their MAO-A and MAO-B inhibiting potency (estimated in in vitro assays) of 1: greater than or equal to 1000 and 1:500, respectively, whereas the least selective drug is cimoxatone with a ratio of 1:66. If in vitro findings are considered, the most potent MAO-A inhibitor seems to be cimoxatone and the least potent toloxatone. After oral administration, however, cimoxatone, brofaromine and moclobemide appear to be about equally effective in inhibiting brain MAO-A. All these drugs are short acting and their MAO-inhibiting action is reversible within 24 h or, in the case of brofaromine, 48 h. Due to the selectivity and reversibility of the MAO-A inhibition, they were found to be less likely to induce large increases in the pressor responses to tyramine, both in animals and in man. In patients treated with 150 mg/day for 4 weeks, brofaromine, which appears to be the best-characterized drug in this respect, produced about a 4 fold increase in sensitivity to tyramine. Although the clinical antidepressant efficacy of these drugs remains to be confirmed in more extensive studies, based on their neurobiochemical and pharmacological properties, clear advantages in respect to their side-effect profiles are to be expected in comparison with the classical MAOIs. PMID- 3045799 TI - Unsolved problems in the pharmacotherapy of depression. AB - Five unsolved problems in the pharmacotherapy of depression are discussed: (a) it is not possible to differentiate endogenous and nonendogenous depression; (b) a selective efficacy of serotonin and noradrenaline reuptake inhibitors cannot be demonstrated; (c) the relationship between plasma levels and antidepressant effect is still unclear: plasma levels are influenced by pharmacogenetic factors, age, route of application, and concomitant treatment with other drugs; (d) evidence is growing for the development of tolerance towards therapeutic effects of antidepressants; (e) no pretreatment variable allows prediction of treatment response: the best predictor is the initial response to treatment. PMID- 3045801 TI - Test models and new directions in dementia research. AB - Psychopharmacology research in the dementia field is particularly difficult because there are no effective treatments on which to base models. Much effort is devoted to attempts to replace defective brain chemical transmitters or neuropeptides, with particular emphasis on the cholinergic system. There is also interest in noradrenaline, serotonin and somatostatin deficits. New directions include the study of glutamate-related excitotoxins such as quinolinic acid as possible causative agents for cerebral degeneration in dementia. Glutamate receptor antagonists, e.g. MK-801, may have the potential to limit or slow down neurodegenerative processes. Neurotrophic factors, e.g. nerve growth factor, may also possess the ability to protect neurons from irreversible loss. PMID- 3045800 TI - Dementia: classification and aspects of treatment. AB - The syndrome dementia is defined by evidence of a decline in memory, thinking, and emotional functions. In this article, dementia is delimited only by its symptoms; the etiology, the course, and the extensiveness of the disorder are not used as criteria of a dementia syndrome. In the descriptive diagnoses of dementia disorders, however, different aspects of the disorders are used. Descriptive diagnoses are (a) benign senile forgetfulness, (b) primary degenerative disorders, (c) secondary dementias, (d) cerebrovascular disorders with dementia, (e) reversible dementias, and (f) dementias with unknown etiology. Primary degenerative disorders include the subgroups (a) senile dementia of Alzheimer type (SDAT), (b) Alzheimer's disease (AD), (c) Pick's disease, and (d) Huntington's chorea. The question whether SDAT a nd AD should be included in one group is discussed. Very interesting and stimulating treatment research is going on in the field of dementias. Although there has still been no real breakthrough, some results indicate that dementia symptoms may be influenced by treatment. At present, treatment strategies go along the following lines: (a) pharmacological treatment, (b) control of sleep apneas and circulatory disturbances, (c) brain tissue transplantation, and (d) psychotherapy and mental activation. The pharmacologyical manipulation of dementia disorders focuses on substituting failing neurotransmitter systems. The use of nootropic drugs and gangliosides is still at an experimental level. Findings of vitamin B12 deficiency in late-onset dementia make vitamin B12 substitution relevant. White matter disturbances such as incomplete infarctions are reported in brains from demented patients. Coincident findings of increased sleep apneas with severe hypoxemia make studies of circulatory disturbances of interest.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3045802 TI - PET scanning--a new tool in clinical psychopharmacology. AB - Quantitative methods were developed for the determination of dopamine and benzodiazepine receptor characteristics in the living human brain by positron emission tomography (PET). As ligands, the 11C-labelled analogues of the selective antagonists of dopamine receptor subtypes, SCH 23390 and raclopride, and the benzodiazepine antagonist, Ro 15-1788, were used. Tracer amounts of the ligands were injected intravenously into healthy volunteers and schizophrenic patients. The distribution of ligand indicated high densities of D1 as well as D2 dopamine receptors in the basal ganglia. Binding of [11C]-SCH 23390 was also significant in the neocortex where it was shown to represent binding to D1 as well as to 5-HT2 serotonin receptors. High densities of specific benzodiazepine receptor binding were obtained in most neocortical brain areas and in the cerebellum. Using saturation procedures, Bmax and Kd values could be obtained for D2 and benzodiazepine receptors. A comparison of D2 receptor densities in drug naive schizophrenic patients and healthy volunteers demonstrated similar receptor characteristics in the major basal ganglia in these groups of subjects. Different chemical classes of conventional and unconventional antipsychotic drugs produced a 65%-85% occupancy of D2 receptors when given in clinical doses to schizophrenic patients. High does of diazepam produced a marked occupancy of benzodiazepine receptors during the first hours after oral administration to healthy volunteers. These in vivo methods should be valuable tools for the further analysis of the effects of drug on neuroreceptors in the living brain of neuropsychiatric patients. PMID- 3045803 TI - Reflections on the history of psychopharmacology. PMID- 3045804 TI - Treatment of cross-bite in early mixed dentition. PMID- 3045805 TI - Formation of stable transcription complexes as assayed by analysis of individual templates. AB - Conditions were established where transient transfection of two marker genes resulted in the expression of one or the other, but not both, in individual cells as assayed by immunofluorescence. Thus, the expression from a single cell reflects the activity of single active transcription templates. Under these conditions, a vector encoding the simian virus 40 large tumor antigen (SV40 T-Ag) driven by the SV40 enhancer and early promoter was transfected into CV-1, L, or HeLa cells yielding, for all three cell types, about 10-30% T-Ag-positive cells as assayed by immunofluorescence. Similar vectors containing either mutated or deleted SV40 enhancers also gave T-Ag-positive cells, but at about 1/100 the frequency. Quantitative analysis showed that T-Ag-positive cells produced about the same amount of T-Ag whether or not an active enhancer was present. Chloramphenicol acetyltransferase-encoding vectors gave the same result. The data are consistent with the hypothesis that at a low, but finite, probability, fully functional transcription complexes can form on a given active template in the absence of enhancer DNA. Enhancers seem to increase the number of active templates. Subcloning experiments suggest that these transcription complexes can be surprisingly stable. PMID- 3045806 TI - Structure of the carboxyl terminus of the RAS gene-encoded P21 proteins. AB - The three-dimensional structures of the carboxyl-terminal regions of the P21 protein products of the human Harvey (Ha), Kirsten (KiA and KiB), and neuroblastoma (N) RAS oncogenes and various mutants have been determined by using conformational energy analysis. The carboxyl-terminal region of P21 has been strongly implicated in the binding of the protein to the inner surface of the plasma membrane without which the protein is inactive. The only invariant residue in this region is Cys-186, which is necessary for the post-translational addition of palmitic acid. The surrounding sequences of the active native proteins differ considerably. Nevertheless, certain amino acid substitutions in this region are known to eliminate membrane binding and protein activity, suggesting that there is a conserved common structural feature in this region in the native proteins that is disrupted in the mutant proteins. Conformational energy analysis shows that the four native P21 proteins have a common structure in the form of an alpha helix for the terminal pentapeptide. A mutant, pBW277, that fails to bind to the membrane and is inactive cannot adopt an alpha-helical structure in this region because of a proline at position 188. Another mutant, pBW766, that retains membrane binding and activity, on the other hand, retains the preference for an alpha-helical conformation in the terminal pentapeptide. These findings suggest that, despite various amino acid sequences in this region, the carboxyl-terminal pentapeptides of the P21 proteins form a distinctive structural domain that must have an alpha-helical structure for membrane binding and intracellular activity. PMID- 3045807 TI - Protein engineering of antibody binding sites: recovery of specific activity in an anti-digoxin single-chain Fv analogue produced in Escherichia coli. AB - A biosynthetic antibody binding site, which incorporated the variable domains of anti-digoxin monoclonal antibody 26-10 in a single polypeptide chain (Mr = 26,354), was produced in Escherichia coli by protein engineering. This variable region fragment (Fv) analogue comprised the 26-10 heavy- and light-chain variable regions (VH and VL) connected by a 15-amino acid linker to form a single-chain Fv (sFv). The sFv was designed as a prolyl-VH-(linker)-VL sequence of 248 amino acids. A 744-base-pair DNA sequence corresponding to this sFv protein was derived by using an E. coli codon preference, and the sFv gene was assembled starting from synthetic oligonucleotides. The sFv polypeptide was expressed as a fusion protein in E. coli, using a leader derived from the trp LE sequence. The sFv protein was obtained by acid cleavage of the unique Asp-Pro peptide bond engineered at the junction of leader and sFv in the fusion protein [(leader)-Asp Pro-VH-(linker)-VL]. After isolation and renaturation, folded sFv displayed specificity for digoxin and related cardiac glycosides similar to that of natural 26-10 Fab fragments. Binding between affinity-purified sFv and digoxin exhibited an association constant [Ka = (3.2 +/- 0.9) x 10(7) M-1] that was about a factor of 6 smaller than that found for 26-10 Fab fragments [Ka = (1.9 +/- 0.2) x 10(8) M-1] under the same buffer conditions, consisting of 0.01 M sodium acetate, pH 5.5/0.25 M urea. PMID- 3045808 TI - Autocrine growth induced by the insulin-related factor in the insulin-independent teratoma cell line 1246-3A. AB - An insulin-independent teratoma-derived cell line, called 1246-3A, has been isolated from the adipogenic cell line 1246, which stringently requires insulin for proliferation. The 1246-3A cell line, which can proliferate in the absence of exogenous insulin, produces in its conditioned medium a growth factor similar to pancreatic insulin by its biological and immunological properties. This factor, called "insulin-related factor" (IRF), was purified and iodinated to study its binding to cell surface receptors. 125I-labeled IRF binding to intact 1246-3A cells is lower than to 1246 cells. Cell surface binding can be restored by culturing the 1246-3A cells in the presence of an anti-porcine insulin monoclonal antibody or by acid prewash of the cells prior to performing the binding. Scatchard analysis of binding indicates that IRF secreted by the 1246-3A cells partially occupies high-affinity binding sites on the producer cells. Moreover, insulin monoclonal antibody inhibits the proliferation of the IRF-producing 1246 3A cells, suggesting that these cells are dependent on the secreted IRF for growth in culture. We conclude that the insulin-related factor secreted by the insulin-independent 1246-3A cells stimulates their proliferation in an autocrine fashion. PMID- 3045809 TI - The role of lipids in Plasmodium falciparum invasion of erythrocytes: a coordinated biochemical and microscopic analysis. AB - The role of lipids in Plasmodium falciparum invasion of erythrocytes was investigated by biochemical and fluorescent microscopic analysis. Metabolic incorporation of radioactive oleate or palmitate and fractionation of radiolabeled phospholipids by thin-layer chromatography revealed no difference in the major phospholipid classes of schizonts and early ring forms after merozoite invasion. Fluorescent anthroyloxy derivatives of oleate and palmitate were also metabolically incorporated into parasite phospholipids. By microscopic analysis, the fluorescent phospholipids were seen localized in the plasma membrane and, within the merozoite, concentrated near the apical end. During invasion fluorescent phospholipid appeared to be injected from the apical end of the merozoite into the host membrane, both within and outside the parasite-host membrane junctions. After invasion fluorescent lipid was only found in the parasite plasma membrane and/or parasitophorous vacuole membrane. Parallel experiments with a fluorescent cholesterol derivative, incorporated into parasite membranes by exchange, revealed neither heterogeneous distribution of label within the parasite nor evidence for cholesterol transfer from merozoite to host cell membrane. Results suggest that during invasion no major covalent alteration of parasite lipids, such as lysophospholipid formation, occurs. However, invasion and formation of the parasitophorous vacuolar membrane apparently involves insertion of parasite phospholipids into the host membrane. PMID- 3045810 TI - Host function of MAK16: G1 arrest by a mak16 mutant of Saccharomyces cerevisiae. AB - The MAK16 gene was first defined as a gene whose mutation resulted in loss of M1 double-stranded RNA virus-like particles. The mak16-1 mutation also produces temperature-sensitive cell growth. We report here that mak16-1 cells arrest at the nonpermissive temperature in G1 phase, such that they are mating competent. We sequenced the MAK16 gene and found an open reading frame of 306 amino acids encoding a predicted protein of Mr 35,694. Two typical nuclear localization signal sequences were found. MAK16-LacZ fusion proteins that include one of these putative signals entered the nucleus, while unfused beta-galactosidase did not, as judged by subcellular fractionation experiments. In the C-terminal third of the MAK16 open reading frame is an acidic region in which 25 of 41 residues are either glutamate or aspartate. This region contains potential phosphorylation sites for "casein kinases," protein kinases specific for serine or threonine residues in an acidic environment. PMID- 3045811 TI - Physical mapping of large DNA by chromosome fragmentation. AB - A technique is described for physically positioning any cloned DNA on a native or artificial Saccharomyces cerevisiae chromosome. The technique involves splitting a chromosome at a specific site by transformation with short linear molecules containing the cloned DNA at one end and telomeric sequences at the other. Recombination between the end of the linear molecules and homologous chromosomal sequences gives rise to chromosome fragments comprising all sequences distal or proximal to the mapping site depending on the orientation of the cloned DNA. The recombinant products are recovered by screening for stabilization of a suppressor tRNA on the linear molecules using a colony color assay. The cloned DNA is positioned relative to the chromosome ends by sizing the chromosomal fragments using alternating contour-clamped homogeneous electric field gel electrophoresis. Application of this technique to organisms other than S. cerevisiae and to the analysis of exogenous DNA cloned in yeast is discussed. PMID- 3045812 TI - Essential fatty acid deficiency prevents multiple low-dose streptozotocin-induced diabetes in CD-1 mice. AB - Multiple i.p. injections of low-dose streptozotocin (40 mg/kg) produce insulitis, beta cell destruction, and diabetes in male CD-1 mice. Recent data also suggest that macrophages figure in the low-dose streptozotocin model. Because other recent studies have shown that essential fatty acid deficiency prevents autoimmune nephritis in mice, decreases the number of resident Ia-positive glomerular macrophages, and decreases the elicitation of macrophages into the glomerulus in inflammation, we examined the effect of essential fatty acid deficiency on the incidence and severity of insulitis and diabetes in CD-1 mice treated with low-dose streptozotocin. Streptozotocin-treated mice on the control diet uniformly developed diabetes (19/19). Essential fatty acid-deficient mice treated with streptozotocin did not develop diabetes (1/13). Mean plasma glucose levels for the control and essential fatty acid-deficient mice were 384.5 +/- 23.6 and 129.1 +/- 15.5 mg/dl, respectively, at the end of 1 month. To discern whether essential fatty acid deficiency prevented the streptozotocin-induced beta cell injury or the inflammatory response to injured beta cells, mice were repleted with daily injections of 99% pure methyl linoleate beginning 3 days after the last streptozotocin injection. These mice also quickly developed severe (3/4) or mild (1/4) diabetes. Histologic examination of the pancreata of control mice or repleted mice showed marked insulitis and beta cell destruction; in contrast, the pancreata of essential fatty acid-deficient mice showed preservation of beta cells and only focal mild peri-insulitis. Essential fatty acid deficiency thus prevents the insulitis and resultant diabetes in low-dose streptozotocin-treated CD-1 mice, suggesting a central role for macrophages and lipid mediators in this autoimmunity model. PMID- 3045813 TI - Biosynthesis of a protein containing a nonprotein amino acid by Escherichia coli: L-2-aminohexanoic acid at position 21 in human epidermal growth factor. AB - Endeavoring to develop a method to biosynthesize proteins substituted with nonprotein amino acids, we attempted the incorporation of L-2-aminohexanoic acid (Ahx) into human epidermal growth factor (hEGF). Escherichia coli YK537 strain harboring plasmid pTA1522, which has the phoA promoter-phoA signal peptide-hEGF gene, was used. Cells were cultured first in high-phosphate medium and then, for induction of the hEGF-encoding gene, transferred to low-phosphate medium containing Ahx (0.25 mg/ml). hEGF and Ahx-substituted hEGF, [Ahx21]hEGF, secreted into the periplasm were recovered. After treatment with H2O2, [Ahx21]-hEGF was clearly separated from methionine-oxidized hEGF by one-step reverse-phase HPLC. Substitution of the methionine residue of hEGF with Ahx was confirmed by the amino acid analysis of [Ahx21]hEGF. The three biological activities of [Ahx21]hEGF were the same as those of hEGF. From the successful production of [Ahx21]hEGF, a basic strategy was established for preparing proteins substituted with nonprotein amino acid (alloprotein). Induction of the phoA promoter of pho regulon and secretion of the product to the periplasm may depress heat shock-like responses and subsequent hydrolysis of the product by cytoplasmic protease. PMID- 3045814 TI - Integration host factor interacts with the DNA replication enhancer of filamentous phage f1. AB - We present data which show that the Escherichia coli integration host factor (IHF) is an activator of phage f1 DNA replication. Phage f1 poorly infects bacterial strains lacking IHF because IHF is required for efficient expression of F-pili, the receptor for f1 phage. However, when F- strains are transfected with f1 DNA the phage replicates in IHF mutants (himA, himD, or himA himD) at a rate of only 3% of that in wild-type bacteria. A plasmid dependent on the f1 replicon fails to transform IHF mutants. By gel retardation analysis, we show that IHF specifically binds to the origin of replication. DNase I "footprinting" experiments demonstrate that IHF binds to multiple sites within the replication enhancer sequence, a cis-acting, A + T-rich sequence that potentiates f1 DNA replication. Moreover, the effect of IHF mutation on f1 growth is suppressed by initiator protein (f1 gene II) mutations that restore efficient replication from origins that lack a functional replication enhancer sequence. This genetic evidence supports the conclusion that the replication enhancer sequence is the site of action of IHF. PMID- 3045816 TI - Selection of the mRNA translation initiation region by Escherichia coli ribosomes. AB - Two genes specifying model mRNAs of minimal size and coding capacity, with or without the Shine-Dalgarno (SD) sequence, were assembled, cloned, and transcribed in high yields. These mRNAs, as well as synthetic polynucleotides, phage MS2 RNA, and a deoxyoctanucleotide complementary to the 3' end of 16S rRNA were used to study the mechanism of translation initiation in vitro. Escherichia coli 30S ribosomal subunits interact with all these nucleic acids, albeit with different affinities; the affinity for the mRNA with the SD sequence (Ka approximately 2 x 10(7) M-1) is more than an order of magnitude higher than that for the mRNA lacking this sequence. The initiation factors are equally required, regardless of the presence of the SD sequence, for 30S and 70S initiation complex formation and for mRNA translation, but the initiation factors do not affect the SD interaction or the binding of the mRNAs to the ribosomes. The SD interaction is also mechanistically irrelevant for 30S initiation complex formation and is not essential for translation in vitro or for the selection of the mRNA reading frame. It is suggested that the function of the SD interaction is to ensure a high concentration of the initiation triplet near the ribosomal peptidyl-tRNA binding site, whereas the selection of the translational start is achieved kinetically, under the influence of the initiation factors, during decoding of the initiator tRNA. PMID- 3045815 TI - Plasmodium falciparum gene encoding a protein similar to the 78-kDa rat glucose regulated stress protein. AB - Genes homologous to heat shock protein 70 have been described in parasitic protozoa. It has been proposed that they may be important to the parasite as it moves from the vertebrate host at 37 degrees C to the insect. We now describe a genomic DNA clone isolated from Plasmodium falciparum that encodes a protein similar in sequence to a mammalian heat shock-related protein, the 78-kDa glucose regulated protein of rat and hamster. The gene is expressed during the erythrocytic stage in both asexual and sexual parasites (RNA blot analysis) and a 72-kDa protein is immunoprecipitated from erythrocytic stage parasites. Importantly, the sequence of the clone is similar to the canonical sequence at the carboxyl termini of glucose-regulated proteins of mammals that determines their localization within endoplasmic reticulum. Since the parasite sequence has only three (Asp-Glu-Leu) of the four carboxyl-terminal amino acids, its location and its function within the parasite remain to be determined. PMID- 3045818 TI - Immunoregulatory activities of human immunodeficiency virus (HIV) proteins: effect of HIV recombinant and synthetic peptides on immunoglobulin synthesis and proliferative responses by normal lymphocytes. AB - Recombinant and synthetic peptides corresponding to envelope proteins of the human immunodeficiency virus (HIV) were examined for their effects on the activities of lymphocytes from normal donors in vitro. Although lymphocytes cultured with env-gag peptides produced significant amounts of IgG, addition of env-gag peptides to a pokeweed mitogen-induced B-cell activation system resulted in suppression of immunoglobulin synthesis by normal lymphocytes. Recombinant antigens, env-gag and env-80 dihydrofolate reductase (DHFR), produced a substantial proliferative response by peripheral blood mononuclear cells (PBMC) as determined by [3H]thymidine incorporation. PBMC precultured with HIV synthetic peptide env 578-608 also manifested significant proliferative responses as compared to control cultures. CD3+ lymphocytes precultured with recombinant HIV antigens, env-gag and env-80 DHFR, and synthetic HIV peptide, env 487-511, showed moderate but significant proliferative responses. Both recombinant antigens and synthetic peptides also produced a dose-dependent stimulatory effect on proliferation by CD3- lymphocytes. Stimulation of CD3+ and CD3- lymphocyte subpopulations induced by env-gag peptides was specifically inhibited by goat anti-env-gag polyclonal antibodies, demonstrating the specificity of the reaction. These studies demonstrate that recombinant and synthetic peptides of the HIV genome express immunoregulatory T- and B-cell epitopes. Identification of unique HIV epitopes with immunogenic and immunoregulatory activities is necessary for the development of an effective vaccine against HIV infection. PMID- 3045817 TI - Ligand-induced desensitization of B-cell membrane immunoglobulin-mediated Ca2+ mobilization and protein kinase C translocation. AB - Binding of ligand to B-cell membrane immunoglobulin (mIg) can lead to activation of a number of distinct biologic responses, including altered expression of genes encoding c-fos, c-myc, and Ia, as well as proliferation and immunologic tolerance. Tolerance could reflect a functional uncoupling of receptors from systems that generate intracellular second messengers (i.e., receptor desensitization). To better understand the molecular basis of immune regulation, we examined the ability of mIg to function as a signal transducer after the cell's initial contact with mIg-binding ligand. The results show that ligand binding to as little as 2-10% of mIgM or mIgD renders the cell unresponsive to ligand binding to the reciprocal isotype as judged by Ca2+ mobilization and protein kinase C translocation responses. This heterologous receptor desensitization lasts longer than 24 hr and does not reflect loss of receptor from the cell surface. Studies with the calcium ionophore ionomycin, 1,2 dioctanoyl-sn-glycerol, and the protein kinase inhibitor staurosporine indicate that both protein kinase C-dependent and protein kinase C-independent (staurosporine-insensitive) mechanisms mediate heterologous desensitization after mIg crosslinking. PMID- 3045819 TI - Gonadotropin-releasing hormone-induced Ca2+ transients in single identified gonadotropes require both intracellular Ca2+ mobilization and Ca2+ influx. AB - We examined the effects of gonadotropin-releasing hormone (GnRH) on the intracellular free Ca2+ concentration ([Ca2+]i) in single rat anterior pituitary gonadotropes identified by a reverse hemolytic plaque assay. Concentrations of GnRH greater than 10 pM elicited increases in [Ca2+]i in identified cells but not in others. In contrast, depolarization induced by 50 mM K+ increased [Ca2+]i in all cells. Ca2+ transients induced by GnRH exhibited a complex time course. After an initial rapid rise, the [Ca2+]i fell to near basal levels only to be followed by a secondary extended rise and fall. Analysis of the Ca2+ transients on a rapid time base revealed that responses frequently consisted of several rapid oscillations in [Ca2+]i. Removal of extracellular Ca2+ or addition of the dihydropyridine Ca2+-channel blocker nitrendipine completely blocked the secondary rise in [Ca2+]i but had no effect whatsoever on the initial spike. Nitrendipine also blocked 50 mM K+-induced increases in [Ca2+]i in identified gonadotropes. The secondary rise induced by GnRH could be enhanced by a phorbol ester in a nitrendipine-sensitive fashion. Multiple spike responses to GnRH stimulation of the same cell could only be obtained if subsequent Ca2+ influx was permitted either by allowing a secondary rise to occur or by producing a Ca2+ transient by depolarizing the cells with 50 mM K+. It therefore appears that the response to GnRH consists of an initial phase of Ca2+ mobilization, probably mediated by inositol trisphosphate, and a subsequent phase of Ca2+ influx through nitrendipine-sensitive Ca2+ channels that may be activated by protein kinase C. The relative roles of these phases in the control of gonadotropin secretion are discussed. PMID- 3045820 TI - Human immunodeficiency virus has an aspartic-type protease that can be inhibited by pepstatin A. AB - The protease encoded by the human immunodeficiency virus (HIV) processes the viral gag and gag-pol protein precursor by posttranslational cleavage. In this study we have demonstrated by site-specific mutagenesis (Asp----Thr) and by pepstatin A inhibition that the recombinant HIV protease is an aspartic-type protease. Furthermore, incubation of HIV-infected H9 cells with pepstatin A inhibited part of the intracellular processing of the HIV gag protein yet had no apparent toxicity on HIV-infected cells during 48 hr of incubation. PMID- 3045821 TI - Discrimination between glutaminyl-tRNA synthetase and seryl-tRNA synthetase involves nucleotides in the acceptor helix of tRNA. AB - Analysis of the in vivo amber suppressor activity of mutants derived from two Escherichia coli serine tRNAs shows that substitution of 2 base pairs in the acceptor helix changes a serine suppressor tRNA to an efficient glutamine acceptor. Determination of the amino acid inserted in vivo into protein by this tRNA shows that these changes reduce the tRNA recognition by seryl-tRNA synthetase while increasing that of glutaminyl-tRNA synthetase. This implies that misaminoacylation in vivo is dependent on the competition by different synthetases for the tRNA. In addition, the "translational efficiency" of tRNA is an integral part in observing misaminoacylation in vivo. PMID- 3045823 TI - Tryptophan repressor of Escherichia coli shows unusual thermal stability. AB - Differential scanning calorimetry demonstrates that the tryptophan repressor of Escherichia coli is unusually resistant to thermal denaturation. The dimeric protein undergoes reversible dissociative unfolding at pH 7.5 centered at about 90 degrees C. The thermal stability may be due in part to the unusual structure of the protein, which is composed of two identical intertwined polypeptide chains. PMID- 3045822 TI - Activation of the human "beta 2-interferon/hepatocyte-stimulating factor/interleukin 6" promoter by cytokines, viruses, and second messenger agonists. AB - The hallmark of "beta 2-interferon (IFN-beta 2)/hepatocyte-stimulating factor/interleukin 6" gene expression is its inducibility in different types of human cells (fibroblasts, monocytes, epithelial cells, and endothelial cells) by different stimuli, which include cytokines such as tumor necrosis factor, interleukin 1 (IL-1) and platelet-derived growth factor, different viruses, and bacterial products such as endotoxin. The activation by cytokines, viruses, and second messenger agonists of the IFN-beta 2 promoter linked to the bacterial chloramphenicol acetyltransferase (CAT) gene was studied after transfection into HeLa cells. A chimeric gene containing IFN-beta 2 DNA from -1180 to +13 linked to the CAT gene was inducible approximately 10-fold by phorbol 12-myristate 13 acetate (PMA), followed, in decreasing order, by pseudorabies and Sendai viruses (7- to 11-fold each); serum (6- to 9-fold); the cytokines tumor necrosis factor, IL-1, and epidermal growth factor (3- to 5-fold each); the cAMP agonists BrcAMP and forskolin and the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (2- to 6-fold each); poly(I).poly(C) (2- to 4-fold); 1,2-diacylglycerol and the calcium ionophore A23187 (1.5- to 2-fold each). Bacterial endotoxin did not activate this IFN-beta 2/CAT fusion gene in HeLa cells. Deletion of the 5' boundary of the IFN-beta 2 DNA from -1180 to -596 in the fusion gene preserved its activation by IL-1, tumor necrosis factor, epidermal growth factor, serum, pseudorabies, and Sendai viruses and by PMA, Br-cAMP, and forskolin; deletion to 225 led to a small reduction (by a factor of 1.5-2) in the responsiveness to serum, PMA, and Sendai virus but not to the other inducers; a further deletion to -112 greatly reduced all responsiveness. Thus, the region between -225 and -113 in IFN-beta 2, which contains DNA motifs similar to the regulatory elements in the human c-fos gene, appears to contain the major cis-acting regulatory elements responsible for the activation of the IFN-beta 2 promoter by several different cytokines, viruses, and second messenger agonists. PMID- 3045824 TI - Cell adhesion induces expression of growth-associated genes in suspension arrested fibroblasts. AB - A methylcellulose suspension system that prevents cell-surface contact with the substrate was used to study the role of cell adhesion in the regulation of proliferation. The nonadhesive conditions established by suspension culture cause BALB/c 3T3 (A31) cells to enter a G0 state of growth arrest within 48 hr as defined by an inhibition of DNA synthesis and a suppression of c-myc and histone mRNA expression. The adhesion of these suspension-arrested cells rapidly induces c-fos, c-myc, and actin gene expression. This stimulation did not depend on the presence of serum since the adhesion of suspension-arrested cells, in the absence of serum, also induced the expression of c-fos and c-myc mRNAs. In addition, adhesion onto fibronectin increased the number of cells able to respond to epidermal growth factor and insulin and progress into S phase. These results indicate that adhesion of suspension-arrested cells activates the G0/G1 transition independent of growth factors. PMID- 3045825 TI - Yeast gene RAD52 can substitute for phage T4 gene 46 or 47 in carrying out recombination and DNA repair. AB - The RAD52 gene of Saccharomyces cerevisiae and genes 46 and 47 of bacteriophage T4 are essential for most recombination and recombinational repair in their respective organisms. The RAD52 gene was introduced into expression vectors that were used to transform Escherichia coli. The expression of RAD52 was then induced, and the ability of RAD52 to complement phage mutants defective in gene 46 or 47 was determined with respect to the three criteria of phage growth, recombination, and recombinational repair. RAD52 gene expression was found to allow growth of gene 46 and 47 mutants under otherwise restrictive conditions, as measured by plaque formation and burst size. Expression of the RAD52 gene also restored the ability of gene 46 and 47 mutants to undergo recombination of rII markers. Furthermore, the RAD52 gene restored the ability of gene 46 and 47 mutants to undergo recombinational repair after UV irradiation. The published DNA sequence of gene RAD52 was compared with the published sequences of genes 46 and 47. Although overall sequence similarities were only marginally significant, RAD52 and gene 46 had substantial sequence similarity over a limited region. PMID- 3045826 TI - Generation of lymphokine-activated killer cells: synergy between tumor necrosis factor and interleukin 2. AB - Large granular lymphocytes (LGL) can be activated by interleukin 2 (IL-2) to lymphokine-activated killers (LAK). The effect of tumor necrosis factor (TNF) on LAK generation was investigated. TNF was found to act synergistically with low concentrations of IL-2 (0.10-0.25 ng/ml), which were ineffective by themselves in inducing LAK activity, to promote the differentiation of LGL into non-major histocompatibility complex-restricted killers. When IL-2 was used at concentrations optimal for LAK generation, TNF did not further enhance this phenomenon. Specific binding of 125I-labeled TNF to LGL was increased by IL-2 stimulation. Scatchard analysis of TNF binding revealed the existence of two classes of binding sites with markedly different affinities (Kd values of 57 and 600 pM). We also demonstrated that the IL-2/TNF synergistic induction of LAK activity did not involve either IL-1 or interferon-gamma. This IL-2/TNF synergistic effect was blocked by anti-Tac antibodies. Immunofluorescence analysis revealed that IL-2/TNF selectively up-regulated Tac antigen expression on LAK precursors. Our results suggest a functional interaction between IL-2 and TNF on LAK precursors, which results in a reduction of the IL-2 concentration required for differentiation of LGL into LAK killers. PMID- 3045827 TI - Primary structure, synthesis, and biological activity of rat endothelin, an endothelium-derived vasoconstrictor peptide. AB - Endothelin is a potent vasoconstrictor/pressor peptide, which we recently characterized from the conditioned culture medium of porcine aortic endothelial cells. We report here the cloning and partial sequencing of the rat endothelin gene. The nucleotide sequence predicted a 21-residue peptide similar to, but distinct from, porcine endothelin; 15 residues of rat endothelin were identical and 3 residues were substitutions by chemically similar amino acid residues to those in the porcine peptide. Synthetic rat endothelin was then prepared according to its deduced amino acid sequence. This synthetic peptide had (i) potent vasoconstrictor activity in the rat aortic strip and in perfused rat heart and (ii) a characteristically long-lasting in vivo pressor activity by intraaortic bolus injection in the conscious rat. PMID- 3045830 TI - Characterization of the trans-Golgi network in BHK cells. PMID- 3045829 TI - Psychological aspects of cancer pain. PMID- 3045828 TI - Genetic exchange among natural isolates of bacteria: recombination within the phoA gene of Escherichia coli. AB - An 1871-nucleotide region including the phoA gene (the structural gene encoding alkaline phosphatase, EC 3.1.3.1) was cloned and sequenced from eight naturally occurring strains of Escherichia coli. Alignment with the sequence from E. coli K 12 made apparent that there were 87 polymorphic nucleotide sites, of which 42 were informative for phylogenetic analysis. Maximum parsimony analysis revealed six equally parsimonious trees with a consistency index of 0.80. Of the 42 informative sites, 22 were inconsistent with each of the maximum parsimony trees. The spatial distribution of the inconsistent sites was highly nonrandom in a manner implying that intragenic recombination has played a major role in determining the evolutionary history of the nine alleles. The implication is that different segments of the phoA gene have different phylogenetic histories. PMID- 3045831 TI - Immuno-isolation of hepatocyte membrane compartments using S. aureus cells. PMID- 3045832 TI - Characterization of intestinal membrane vesicles with flow cytometry. PMID- 3045833 TI - Fusion in the endocytic pathway reconstituted in a cell-free system using immuno isolated fractions. PMID- 3045834 TI - Functional characteristics of the endosomal apparatus of rat liver parenchyma. PMID- 3045835 TI - Continuous flow electrophoresis applied to the separation of cells, membranes and membrane functional domains. PMID- 3045836 TI - Immuno-isolation of subcellular components. PMID- 3045837 TI - Immunoaffinity techniques for the purification and functional assessment of subcellular organelles. PMID- 3045838 TI - Prostacyclin does not change during an oxygen induced increase in pulmonary blood flow in the fetal lamb. AB - The role of prostacyclin in mediating the increase in pulmonary blood flow caused by an increase in oxygen tension in the fetal lamb was investigated. Plasma concentrations of 6-keto-PGF1 alpha, the hydrolysis product of prostacyclin, were measured during an increase in pulmonary blood flow caused by a rise in oxygen tension in eight intrauterine fetal lambs. Fetal oxygen tension was increased by placing the pregnant ewes in a hyperbaric chamber and having them breathe 100% oxygen at three atmospheres absolute pressure. This increased fetal PaO2 from 27 +/- 3 to 60 +/- 6 torr (mean +/- S.E., p less than or equal to 0.0001) and increased the proportion of right ventricular output distributed to the fetal lungs from 6 +/- 2 to 45 +/- 7% (mean +/- S.E., p less than or equal to 0.001). However, the fetal plasma concentration of 6-keto-PGF1 alpha did not change, 186 +/- 26 to 208 +/- 40 pg/ml (mean +/- S.E.). Indomethacin decreased plasma concentrations of 6-keto-PGF1 alpha in each of three fetuses but did not decrease the proportion of right ventricular output distributed to their lungs. The increase in pulmonary blood flow caused by an increase in oxygen tension in the fetal lamb is not associated with an increase in plasma concentrations of 6-keto PGF1 alpha. Prostacyclin does not appear to be involved in the increase in pulmonary blood flow caused by the increase in oxygen tension at birth. PMID- 3045840 TI - Endogenous prostanoids and arterial contractility. PMID- 3045839 TI - Cyclosporine A nephrotoxicity--the role of thromboxane A2. PMID- 3045841 TI - Effect of fenfluramine on sympathetic firing rate. AB - The effects of acute and chronic treatment with fenfluramine have been explored in two experiments. Three and twenty-four hours following the injection of fenfluramine 20 mg/kg the firing rate of sympathetic efferent nerves to brown adipose tissue was significantly increased compared to sham injected controls. Body weight loss following acute treatment with fenfluramine was significantly greater at three and twenty-four hours than in the vehicle-treated controls. In the chronic experiment animals were treated once daily for 12 days with 20 mg/kg of fenfluramine. There were two control groups. One control group ate ad lib and a second control group was pair fed to maintain body weight comparable to that of the fenfluramine-treated animals. By the twelfth day food intake in the fenfluramine-treated animals had returned to control levels. Sympathetic firing rate after three days of treatment with fenfluramine was significantly higher in the treated animals than in ad lib fed controls. The ad lib fed controls were likewise significantly higher than the vehicle-treated, pair-gained controls. After 12 days of treatment fenfluramine treated animals had sympathetic firing rates which were still slightly but significantly higher than those of the vehicle-treated controls whereas the vehicle-treated, pair-gained animals had a small but significantly reduced firing rate. These data support the hypothesis that fenfluramine can increase peripheral sympathetic activity. PMID- 3045842 TI - New hypnotic agents: clinical studies in general practice. AB - The type of sleep problem should be determined so that the most appropriate hypnotic can be used, and in this respect duration of action is an important property. The benzodiazepine hypnotics are adequate for this purpose, but problems may arise due to daytime after-effects and the possibility of dependence developing. Non-benzodiazepine hypnotics may be useful alternatives and our group has undertaken double-blind comparative trials with two such compounds, namely zopiclone and zolpidem. Zopiclone was compared to temazepam in a cross-over trial on 36 patients and similar hypnotic effects were recorded. In a parallel group study, zolpidem 10 mg was compared to zolpidem 20 mg and placebo in 88 patients. Both doses of zolpidem were significantly better than placebo on a number of the parameters recorded, side-effects were negligible and there was no evidence of any rebound insomnia during the final control week. PMID- 3045843 TI - [Progress in the field of drug development. 21]. PMID- 3045844 TI - [Research schools in pharmacy. 1. Theory and methods in research schools in pharmacy; 29. The history of pharmaceutical science]. AB - Part 1: On the theory and the methodology of research schools Contributions on the History of pharmaceutical science, part 29. The paper is the first part of an extensive investigation about the development of research schools of pharmacy. The major purpose of this part is to give a definition of pharmaceutical research schools and a methodology for the historical study of research schools. In trying to postulate and analyse the most propitious conditions by which a research school could flourish in the time from the end of the 18th century to the end of the 20th century, we have clearly take into account aspects, chief elements, such as the director or teacher, the scientific programme the students, the research conditions and the public and social recognition. To this aspects is given a methodological-programme for the historical analysis. PMID- 3045845 TI - [Preparation of biological research material for HPLC analysis]. PMID- 3045846 TI - Effect of procaine injection into the ventromedial hypothalamic area (VMH) of the rat on serum insulin, glucose and corticosterone and gastric emptying rate. AB - Bilateral injection of procaine into the VMH resulted in a decrease in serum glucose and immunoreactive insulin levels in both food-deprived rats and meal-fed rats. Corticosterone levels and gastric emptying rate were unaffected by these VMH procaine injections. Since it is likely that the procaine disrupted only those neurons whose cell bodies and synapses lay in the region of the VMH, the results suggest that the two major factors in the etiology of the 'VMH syndrome' following electrolytic lesions, rapid emptying and insulin hyperresponsivity, are dependent upon destruction of fibers of passage through the VMH. Finally, the results further support the proposal that the VMH exerts tonic excitatory control over liver glucose output. PMID- 3045847 TI - [The sleep-wake rhythm of the fetus]. AB - Our work refers to a body of studies carried out by Sonargram observation, which allowed us to study fetal movements in real time. These fetuses were studied not only as parameters of correct neurobiological maturity but also from the psychological interest which looks for the beginning of thinking based on the question: When and how does the thought process begin in biological matter? The study comparing the movement of fetuses and new-born babies to psychoanalytical research (which have been described in other works) led us to formulate the following hypothesis: motor and sense experiences can only become "mental" after separation from the mother; it can only come about by birth which effectively forces a confrontation on biological grounds: it prepares the thinking process without yet having it. There is a debate between these positions and psychoanalytical schools of thought in which thinking could precede the birth of the human fetus; there might be a stage of sleep (REM) that corresponds to adult dreaming. PMID- 3045848 TI - [Nosology and systems for data recording in child and adolescent psychiatry]. AB - Thanks to a system of classification acceptable to a majority of child psychiatrists, recording mental problems has been a necessity since the 1960's. The two main classifications currently being used are that of the World Health Organization (CIM-9) and that of the American Association of Psychiatry (DSM III), both of which categorize, with categories proper to children. There are also classifications dealing with dimensions which show child psychopathology better to psychologists. A system of French classification is being studied. The merits of a good classification are well set out. No existing system of classification has them completely. The CIM-9 and DSM-III are being fine-tuned. Whichever system used, date gathering on a particular patient must be rigorous. Studies carried out in Great Britain and the United States have led to the formulation of semi-structured interviews, an evaluation scale and questionnaires that now cover most child and adolescent psychiatry. PMID- 3045850 TI - [Fancies about fever in ancient medicine]. PMID- 3045851 TI - The nature of scientific discovery. "The limits of science". By Peter Medawar. Essay review. PMID- 3045849 TI - Heart rate and blood pressure during placebo-associated panic attacks. AB - Increases in the anxiety ratings, heart rate, and systolic and diastolic blood pressure were compared in eight panicking and 57 nonpanicking panic disorder patients during placebo (dextrose) infusions. Heart rate and systolic and diastolic blood pressure increased significantly in placebo-infused panickers, but these increases did not correlate well with increases in anxiety. PMID- 3045852 TI - Genes and DNA. "The transforming principle. Discovering that genes are made of DNA." By M. McCarty. Essay review. PMID- 3045853 TI - On the contribution of Volterra and Lotka to the development of modern biomathematics. PMID- 3045854 TI - Laboratory technology and biological knowledge: the Tiselius electrophoresis apparatus, 1930-1945. PMID- 3045855 TI - [Galen's gerontology]. PMID- 3045857 TI - [The epistemological regulation of medicine]. PMID- 3045858 TI - "To improve the evidence of medicine": arithmetic observation in clinical medicine in the eighteenth and early nineteenth centuries. PMID- 3045859 TI - Mind and brain in medical thought during the romantic period. PMID- 3045856 TI - History of discoveries of malaria parasites and of their life cycles. PMID- 3045860 TI - Ideology and health care in Britain: Chadwick to Beveridge. PMID- 3045861 TI - [The concept of disease subjacent to attempts at computerization of medical diagnosis]. PMID- 3045862 TI - The population biology of parasite-induced changes in host behavior. AB - The ability of parasites to change the behavior of infected hosts has been documented and reviewed by a number of different authors (Holmes and Bethel, 1972; Moore, 1984a). This review attempts to quantify the population dynamic consequences of this behavior by developing simple mathematical models for the most frequently recorded of such parasite life cycles. Although changes in the behavior of infected hosts do occur for pathogens with direct life cycles, they are most commonly recorded in the intermediate hosts of parasites with complex life cycles. All the changes in host behavior serve to increase rates of transmission of the parasites between hosts. In the simplest case the changes in behavior increase rates of contact between infected and susceptible conspecific hosts, whereas in the more complex cases fairly sophisticated manipulations of the host's behavioral repertory are achieved. Three topics are dealt with in some detail: (1) the behavior of the insect vectors of such diseases as malaria and trypanosomiasis; (2) the intermediate hosts of helminths whose behavior is affected in such a way as to make them more susceptible to predation by the definitive host in the life cycle; and (3) the behavior and fecundity of molluscs infected with asexually reproducing parasitic flatworms. In each case an expression is derived for R0, the basic reproductive rate of the parasite when first introduced into the population. This is used to determine the threshold numbers of definitive and intermediate hosts needed to maintain a population of the pathogen. In all cases, parasite-induced changes in host behavior tend to increase R0 and reduce the threshold number of hosts required to sustain the infection. The population dynamics of the interaction between parasites and their hosts are then explored using phase plane analyses. This suggests that both the parasite and intermediate host populations may show oscillatory patterns of abundance. When the density of the latter is low, parasite-induced changes in host behavior increase this tendency to oscillate. When intermediate host population densities are high, parasite population density is determined principally by interactions between the parasites and their definitive hosts, and changes in the behavior of intermediate hosts are less important in determining parasite density. Analysis of these models also suggests that both asexual reproduction of the parasite within a host and parasite-induced reduction in host fecundity may be stabilizing mechanisms when they occur in the intermediate hosts of parasite species with indirect life cycles.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3045863 TI - [Improvement of the mechanical retention of adhesive bridges]. PMID- 3045864 TI - [Eight-year study of ceramic post implant as a late implant for single tooth loss (Brinkmann Class I)]. PMID- 3045866 TI - [From alginate and hydrocolloid impression to the model (I)]. PMID- 3045865 TI - [An uncommon case in dental implantology]. PMID- 3045868 TI - [From alginate and hydrocolloid impression to the model (II)]. PMID- 3045867 TI - [Dentistry in ancient China]. PMID- 3045869 TI - [Tooth set-up in wax (II)]. PMID- 3045870 TI - [Care of a one-sided shortened tooth row using FM-hinge joint]. PMID- 3045872 TI - Abstracts: European Symposium 1987. Half body and total body irradiation. Dresden, Sept. 30-Oct. 3, 1987. PMID- 3045871 TI - [The cut model (I). Basic technical work for accurately fitting crowns and bridges]. PMID- 3045873 TI - [Radiologic search for tumors in skin changes. Paraneoplastic syndromes and hereditary diseases with increased risk of tumors]. AB - Cutaneous paraneoplastic syndromes and inherited skin diseases that are known to have an increased tumor risk can be diagnosed by a physician with knowledge of their distinct clinical features. Cutaneous paraneoplastic syndromes are regarded as caused by tumor-cell products and should disappear after sufficient tumor treatment. The risk of malignancy varies in different inherited syndromes, but knowledge of the cutaneous symptoms in these diseases facilitates appropriate diagnostic procedures. PMID- 3045875 TI - [Sensitivity and specificity of sonographic diagnosis of the lymph nodes in malignant melanoma]. AB - A total of 213 melanoma patients were checked perioperatively with a 5-MHz sonographic scanner in order to detect lymph-node metastases; they were also checked in the scope of tumor follow-up. Of the 415 sonographic results, the method yielded a 97% accuracy. The soundness of lymph-node sonography has been proved histologically and/or through clinical observation. Compared to the other diagnostic techniques available for checking surface and subsurface lymph-node groups, lymph-node sonography is an advantageous combination of diagnostic practicability, accuracy, economic feasibility, and patient acceptance. The possible therapeutic implications are also discussed in this paper. PMID- 3045874 TI - [Imaging procedures in systemic connective tissue diseases (collagenoses)]. AB - Systemic lupus erythematosus (SLE) and progressive systemic sclerosis (PSS) represent the most frequent manifestations of systemic connective tissue diseases (collagen diseases). Radiological examinations are employed to estimate the extension and degree of the pathological process. In addition, progression of the disease can be verified. In both of the above collagen diseases, specific radiological findings can be observed that permit them to be differentiated from other entities. An algorithm for the adequate radiological work-up of collagen diseases is presented. PMID- 3045876 TI - [Skeletal changes following long-term treatment with retinoids]. AB - The synthetic retinoids, the vitamin-D-derivatives etretinate and isotretinoin, have substantially enlarged the therapeutic arsenal in dermatology. They are primarily used in severe cases of acne and cornification disorders. In the majority of cases, long-term treatment is necessary. Certain side effects in the skeletal system can occur, e.g., osteoporosis, premature epiphyseal closure, and changes similar to DISH (diffuse idiopathic skeletal hyperostosis). We discuss the reports in the literature and our own observations in 31 patients treated at the Westphalian Wilhelms University in Muenster, as well as at the Technical University in Munich. In 3 out of 31 patients treated by retinoids on a long-term basis, skeletal changes were found radiologically as a result of the retinoid medication. PMID- 3045877 TI - [Pulmonary involvement in tuberous sclerosis]. AB - The authors present a case of tuberous sclerosis with marked pulmonary involvement, confirmed by both radiological and pathological studies. The radiological manifestations and basic pathology of this rare condition are reviewed with emphasis on differential diagnosis. PMID- 3045878 TI - [Ultrasonic morphology of the normal Achilles tendon and pattern of pathological changes]. AB - The anatomical structure of the Achilles tendon causes a fibrillary echo texture with fine, parallel echogenic lines. The peritendineum outlining the tendon can be delineated. Achilles tendon ruptures can easily be detected by ultrasound, as can the structural changes in achillodynia. The typical sonographic patterns of rupture of the Achilles tendon and achillodynia are demonstrated. PMID- 3045879 TI - [From the early history of roentgen documentation]. AB - By chance, eight X-ray plates dating from 1899 were found in 1986. Each of these glass plates shows the patient's name, the date of the examination were performed in Vienna by Prof. Franz Exner, who was a friend of W.C. Roentgen. The glass plates are 25 x 13 cm in area and 3 mm thick. One side is coated with silver bromide. Probably these are some of the earliest X-rays records in the history of radiology. Their quality is comparable with the quality that can be achieved today, even though the materials and the imaging method used were somewhat unsophisticated. PMID- 3045880 TI - Vasodilators in the management of congestive heart failure. PMID- 3045881 TI - Implantable drug delivery devices (pumps, ports) in cancer therapy. PMID- 3045882 TI - Neural substrates of aggression and flight in the cat. PMID- 3045883 TI - Complex-cell receptive field models. PMID- 3045885 TI - Cell death and the elimination of retinal axons during development. PMID- 3045884 TI - Cholinesterase and its molecular forms in pathological states. PMID- 3045886 TI - [Symposium on the humanistic value of the work of Antoni Kepinski]. PMID- 3045887 TI - [Antoni Kepinski as an authority]. PMID- 3045888 TI - [Antoni Kepinski's government of souls]. PMID- 3045889 TI - [A few remarks concerning the dialog on salvation. In memory of Antoni Kepinski]. PMID- 3045890 TI - [Affirmation of life in the philosophy of Antoni Kepinski]. PMID- 3045891 TI - ["Amo, ergo sum" ("I love, therefore I am"). Antoni Kepinski on love]. PMID- 3045892 TI - [Empirical basis of the knowledge of man (a paper dedicated to the memory of Antoni Kepinski)]. PMID- 3045893 TI - [Antoni Kepinski as an intellectual]. PMID- 3045894 TI - [Problem of fear in the works of Antoni Kepinski]. PMID- 3045895 TI - ["Faith, hope and love accompany one another"]. PMID- 3045896 TI - [Bibliography of Antoni Kepinski's sp5entific papers]. PMID- 3045897 TI - Intrahepatic calculi: imaging by MR. AB - The magnetic resonance (MR) examinations of six patients with intrahepatic calculi were reviewed retrospectively to 1) determine the ability of MR to demonstrate intrahepatic calculi and 2) assess the MR appearance of the stones. In five out of six cases, MRI demonstrated intrahepatic calculi. In three cases, stones exhibited a low intensity signal on the different spin echo (SE) sequences, as previously described by in vitro and in vivo studies. However in two other cases, a significant signal with short T1 and relatively long T2 relaxation times was noticed. These different features are discussed in relation to chemical and physical differences in intrahepatic calculi and compared with variable CT attenuation values of stones. MRI seems to provide complementary information concerning intrahepatic calculi. PMID- 3045899 TI - [Nuclear medicine in hepatology]. PMID- 3045900 TI - [Introduction to the experiments of genetic engineering. VII. Chromosome mapping]. PMID- 3045898 TI - Clinical trial of intravenous infusion of bromodeoxyuridine (BUdR) for radiosensitization of malignant brain tumors. AB - Bromodeoxyuridine (BUdR) is a radiosensitizer that can be incorporated into cellular DNA as a substitute for thymidine at the time of DNA synthesis. As the steady-state arterial concentration of BUdR given by means of intravenous infusion was recently presented, the possibility of revival of BUdR as a radiosensitizer administered by the intravenous route was suggested. Based on the experience of BAR therapy and phase-I studies by NIH and UCSF, 12 hours of BUdR at a dose of 800-1,000 mg/m2 for five days a week was given to 23 patients with primary and secondary malignant brain tumors during radiation therapy. Radiation therapy was planned at a weekly dose of 10 Gy for five to six weeks. Fifteen patients received 1,000 mg/m2 of BUdR; six of them tolerated more than three weeks of treatment. In eight patients given doses of 800 mg/m2, five patients tolerated more than three weeks. The most remarkable toxic effects were myelosuppression and stomatitis, which were major obstacles to maintaining the schedule. More than 50% reduction of tumor volume was obtained in five of 12 cases of evaluated gliomas (42%) and three of four cases of metastatic tumors (75%). The median time to tumor progression in seven patients with glioblastoma was 37 weeks. PMID- 3045902 TI - The EPA Science Advisory Board: a ten-year retrospective view. PMID- 3045901 TI - [Combined sciatic/3-in-1 block. III. Prilocaine 1% versus mepivacaine 1%]. AB - In a randomized, double-blind study, the efficacy of prilocaine 1% (group 1, 30 patients) was compared to mepivacaine 1% (group 2, 30 patients). All patients had a combined sciatic/femoral block for surgery of the lower extremities; a tourniquet was applied in each case. In each patient the block was done using 50 ml of a 1% solution of local anesthetic: 20 ml for the sciatic and 30 ml for the 3-in-1 block. The time to onset of sensory block was approx. 4 min in both groups; the onset of motor blockade averaged 6 min and after 10-12 and 15 min sensory and motor blockades, respectively, were complete. On the average, patients were pain-free for 254 min with prilocaine and 267 min with mepivacaine. Four of 30 patients (= 13%) in group 1 an 6 of 30 patients (= 20%) in group 2 had an unsatisfactory blockade and had to be supplemented by analgesics or general anesthesia. The finding of a significant correlation between the voltage necessary for stimulation and the efficacy of the blockade underlines the importance of correct stimulation when identifying the nerves. PMID- 3045903 TI - How do cancer risks predicted from animal bioassays compare with the epidemiologic evidence? The case of ethylene dibromide. AB - Cancer risks for ethylene dibromide (EDB) were estimated by fitting several linear non-threshold additive models to data from a gavage bioassay. Risks predicted by these models were compared to the observed cancer mortality among a cohort of workers occupationally exposed to the same chemical. Models that accounted for the shortened latency period in the gavaged rats predicted upper bound risks that were within a factor of 3 of the observed cancer deaths. Data from an animal inhalation study of EDB also were compatible with the epidemiologic data. These findings contradict those of Ramsey et al. (1978), who reported that extrapolation from animal data produced highly exaggerated risk estimates for EDB-exposed workers. This paper explores the reasons for these discrepant findings. PMID- 3045904 TI - Economic costs of misinforming about risk: the EDB scare and the media. AB - This study reports results of an analysis of consumer responses to news reports of grain-product contamination by the pesticide ethylene dibromide (EDB). The results demonstrate that it is possible to quantify market disruption related to the dissemination of risk information. Implications include the need for increased awareness among risk managers that public perceptions, regardless of their objective accuracy, can induce real economic costs. Such costs should be considered in designing regulatory and information policies. PMID- 3045905 TI - [Metabolic control using a subcutaneous insulin infusion pump: effects 2 years after treatment]. PMID- 3045906 TI - [Localization of the parathyroid glands]. PMID- 3045907 TI - [Gammagraphic and nuclear magnetic resonance studies on the diagnosis of cancer of the thyroid]. PMID- 3045908 TI - [The Guadalajara study: different forms of response of diabetic patients in a course of continuing diabetology education. Preliminary results after a 1-year follow-up]. PMID- 3045909 TI - [Resistance to treatment of Plasmodium falciparum in Central Africa]. PMID- 3045910 TI - [Evaluation of an anti-rubella vaccine through epidemiologic study of an outbreak of the disease in a basic secondary school in the country]. PMID- 3045911 TI - [The SAVE automated epidemiologic surveillance system]. PMID- 3045912 TI - [The predatory capacity of the larvivorous fish Poecilia reticulata (Peters, 1985) (Cyprinodontiformes: Poecillidae) in a natural breeding-ground for the mosquito Culex quinquefasciatus Say, 1823]. PMID- 3045913 TI - [Infestation and parasitic development of Romanomermis culicivorax (Ross and Smith, 1976) (Rhabditida, Mermithidae) in larvae of Anopheles (Nyssorhynchus) albimanus Wiedeman, 1821 (Diptera: Culicidae) under laboratory conditions]. PMID- 3045915 TI - [Predatory capacity of Poecilia (Lebistes) reticulata Peters, 1895 (Cyprinodontiformes: Poecillidae) against larvae of Culex quinquefasciatus Say, 1823 and Aedes aegypti Linnaeus, 1762 (Diptera: Culicidae) under laboratory conditions in Cuba]. PMID- 3045914 TI - [Evaluation and introduction of the indirect immunofluorescence test (toxoplasmosis) in the diagnostic services of the Pedro Kouri Institute of Tropical Medicine and its relation to the complement fixation test]. PMID- 3045916 TI - [Trial of mathematical models for the prediction of the epidemic situation of acute respiratory diseases. Cuba, 1984]. PMID- 3045917 TI - [The 50th anniversary of the founding of the Pedro Kouri Institute of Tropical Medicine. 1937-87]. PMID- 3045918 TI - [The age of cultures of Romanomermis culicivorax (Ross and Smith, 1976) (Rhabditida: Mermithidae) in the infestation of mosquito larvae of the species Culex quinquefasciatus Say, 1823, under laboratory conditions)]. PMID- 3045919 TI - [Current thoughts on the immunology of leprosy]. PMID- 3045920 TI - [Hepato-bronchial fistula: a late complication of liver abscess. Presentation of a case and review of the literature]. PMID- 3045921 TI - [Treatment by drawing lots in a therapeutic trial with a balance distribution of prognostic factors: the minimization method]. AB - In a controlled clinical trial, a more powerful design can be obtained by balancing the treatment groups for the main prognostic factors rather than ignoring these factors in the randomization. The method of random permuted blocks within strata becomes inadequate as the number of prognostic factors increases. Another method known as minimization is described, which balances the marginal distribution of prognostic factors; it was proposed by Taves, and generalized by Pocock and Simon. It avoids the problems arising from an imbalance in a major prognostic factor. PMID- 3045922 TI - Risk factors of morbidity and mortality in surgically treated chronic aortic valvular heart disease. AB - A cohort of 617 patients underwent aortic valve replacement (AVR) the median follow-up time was 3.4 years with a range (0-7.8) years. The incidence of early mortality was 5.0% and the five years survival (77.9 +/- 1.9)%. Risk factors of early mortality and morbidity (Low Output Syndrome) occurring the first 31 days after operation were pinpointed. Another analysis was done to estimate independent predictors of late premature death from all causes, and from specific causes (cardiac related, sudden cardiac). One of the major late morbid events was the appearance of systemic emboli. Its rate was 1.4% pats. Year. Its risk factors were presence of pure aortic regurgitation and advanced age at operation. The relative survival rate was at 5 years of 87.0% for the total cohort, but for our younger patients (age less than 30 years), we reached 99.4%. Our results suggest more aggressive measures to correct the hazard in AVR, and impose carefulness in comparing quality of AVR from different institutions for mortality and morbidity. Finally the results of AVR are rather palliative than curative except for our younger patients where we reached curability. PMID- 3045923 TI - [Ischemic cardiopathy (IV). Physiopathological bases for arrhythmias in ischemia and coronary reperfusion]. PMID- 3045924 TI - Mammalian reproductive strategies: genes, photoperiod and latitude. AB - This paper considers how and why natural selection might promote or block the photoperiodic regulation of a mammal's reproduction. The factors most important in making this decision would seem to be the following: life expectancy, length of the female's cycle, feeding strategy, the presence or absence of survival mechanisms like hibernation, and the nature of the seasonal challenges offered by the mammal's habitat. A speculative scheme is offered for the potential utility of this type of regulation dependent upon life expectancy and latitude of residence. PMID- 3045925 TI - How does melatonin control seasonal reproductive cycles? AB - The pineal gland is essential for the perception of photoperiod change in many species. Information about photoperiod length is conveyed through pineal secretion of the methoxyindole melatonin. Melatonin, suitably administered in physiological quantities is equipotent with artificial photoperiod in the induction of photoperiodic responses. Most experimental work suggests that it is the duration of high night time melatonin secretion (positively correlated with the length of the natural or artificial dark phase) which conveys the photoperiodic signal. Continuous release implants induce short day effects in ewes, entirely comparable to daily feeding of melatonin or short photoperiod. A minimum duration of secretion rather than a specific duration is therefore probably critical to short day effects. There appears to be a seasonal variation in sensitivity to short day melatonin effects (induction of early oestrus) which can be shifted to an earlier time of year following one oestrus advance the previous year. Short duration melatonin is read as a long day even secreted with 22 hour periodicity, suggesting a lack of circadian variation in sensitivity to melatonin. PMID- 3045926 TI - Short- and long-term effects of manipulation of the pineal/melatonin axis in ewes. AB - The experiments which have provided insight into the role of the pineal gland and melatonin in sheep reproduction are reviewed. There is now strong evidence that timed daily melatonin administration and continuous administration via implants are equally effective in promoting the same physiological consequences as short daylength. Thus melatonin treatments lasting six weeks can result in an advance of the breeding season and an advance in the seasonal peak in ovulation rate. Much longer periods of exposure to melatonin or long-term pinealectomy result in a different response. In the second part of this review the long-term effects of pineal manipulation in juvenile and pubertal ewes are compared and contrasted. Five years after pinealectomy (at 7.5 months) ewes displayed normal timed breeding seasons without exhibiting a normal annual pattern of LH sensitivity to oestradiol. Pinealectomy at 2.5 months of age resulted in delayed puberty and breeding seasons out of synchrony with normal ewes two years later. These animals maintained an LH sensitivity to oestradiol consistent with their own breeding season. Melatonin implants which delivered melatonin for over a year caused similar effects as pinealectomy at 2.5 months but had essentially opposite effects to pinealectomy in adults. These results generate many questions about the different perception of the melatonin signal in prepubertal versus adult ewes and the factors involved in the onset and offset of the breeding season. PMID- 3045927 TI - The role of the pineal gland in the photoperiodic control of reproduction in different hamster species. AB - The pineal gland is known to play a central role in the photoperiodic control of reproduction in seasonal breeders. The present review, based on experimental data obtained in four different species of hamster, the Syrian or golden, the Djungarian or Siberian, the Turkish and the European, attempts to evaluate the role of the pineal in this phenomenon, the message which is conveyed from this gland as well as the mechanism of action of this message. Melatonin one of the 5 methoxyindoles rhythmically synthesized in the pineal gland, appears to be the pineal hormone conveying the photoperiodic message. The importance of the duration of the nocturnal peak of circulating melatonin is now well established, but our knowledge on the sites of action and on the mechanisms of action of melatonin is still rather poor. The presently available data suggest that melatonin can act at different levels, either on specific receptors, on receptors of other transmitters, on various molecular processes after diffusion in the cell or indirectly via effects on other endocrine hormones, e.g. gonadal steroids. PMID- 3045928 TI - Characteristics of the melatonin signal that provide the photoperiodic code for timing seasonal reproduction in the ewe. AB - The following is a progress report of our studies to identify important features of the circadian pattern of melatonin secretion which provide the photoperiodic code for daylength in regulating seasonal breeding in the Suffolk ewe. The first series of experiments evaluated two conceptual models of how melatonin codes for daylength: the circadian timing of the melatonin elevation as opposed to the length of the time melatonin is elevated during each 24-hr period (phase vs duration). Strong support has been gathered for the duration hypothesis. No evidence was obtained to support a role for phase; nevertheless, this hypothesis could not be discounted definitively. A second series of studies evaluated the importance of the previous melatonin pattern to the interpretation of a given melatonin signal. Evidence is presented that a fixed melatonin pattern can maintain a given reproductive response only for a limited length of time and that this response can be prolonged by appropriate changes in the melatonin pattern. Thus, change is an important feature of the melatonin signal. Further, the nature of the melatonin change appears to be crucial, specifically whether the nocturnal elevation increases or decreases in duration. Thus, transfer to a common photoperiod can promote either reproductive induction or arrest, depending upon whether the transfer leads to a decrease or increase in daylength. This has important ramifications to the photoperiodic timekeeping process in those species of mammals which utilize daylength to time their seasonal reproductive cycle. PMID- 3045929 TI - Pharmacological manipulation of the mammalian circadian clock: implications for the control of seasonal reproductive cycles. AB - Although artificial control of the light cycle can be used to regulate the seasonal reproductive cycle of many animals under laboratory conditions, such regulation is often not possible in a standard agricultural environment. An alternative strategy for regulating the seasonal reproductive cycle of photoperiodic animals is to use drugs that either mimic the effects of light on reproductive function or that induce an alteration in the way the circadian clock system is entrained by the light cycle. The ability to use drugs to achieve these objectives has been demonstrated in the laboratory, but it remains to be determined if such an approach can be used to regulate the breeding season of farm animals under normal agricultural conditions. PMID- 3045930 TI - [The value of examination of the placenta in the 3d pregnancy trimester]. PMID- 3045931 TI - [Urinary infection in children]. PMID- 3045933 TI - [Vascular pain of the face: diagnosis and differential diagnosis]. PMID- 3045932 TI - [Primary pulmonary sarcomas (review of the literature apropos of 2 case reports)]. PMID- 3045934 TI - [Significance of sonography in the assessment of shoulder symptoms]. PMID- 3045935 TI - [Cervical myeloradiculopathy]. PMID- 3045936 TI - [The biological status of elderly persons]. PMID- 3045937 TI - [The use of ultrasonography in splenic injuries]. PMID- 3045938 TI - [The treatment of type II diabetes]. PMID- 3045939 TI - [Pseudo-allergic rhinitis in sinus aspergillosis. Apropos of a case]. PMID- 3045940 TI - [Secretory otitis: a review]. PMID- 3045941 TI - [Cantonal medical societies in French-speaking Switzerland--societies of French speaking physicians]. PMID- 3045943 TI - Use of light-cured hybrid composite on metal frameworks for use in implant cases. PMID- 3045942 TI - A custom dental ceramic firing support. PMID- 3045945 TI - Remembering yesterday: Kathleen Hodgson. Interview by Gwen Kavanagh. PMID- 3045944 TI - Remembering yesterday: Dorothy D'Arcy-Goldrick. Interview by Jo Russell. PMID- 3045946 TI - Hormonal control of collagen metabolism--Part I. AB - The collagens are a group of the connective tissue proteins widely distributed in human and animal body. The collagens together with other components of the tissue form a complex regulatory system, and are subject to hormonal control. The present paper reviews the effect of thyroid hormone and thyroid-stimulating hormone (thyrotropin) on collagen. Thyroid hormone influences collagen biosynthesis and degradation, and this effect is responsible for various pathophysiological phenomena, including alterations in urinary excretion of hydroxyproline and hydroxylysine, hyperthyroid acropachy, pretibial myxedema, impaired wound repair and other fibrosis-associated processes as well as a tadpole tail resorption in amphibians. Thyrotropin also directly affects the connective tissue, and is responsible for exophthalmos caused by acumulation of certain components of the tissue in retrobulbar space. Collagen fibres are present in the thyroid gland, and take part in the embryogenesis of the gland. PMID- 3045948 TI - Presence of Mycobacterium leprae-reactive lymphocytes in lymph nodes of lepromatous leprosy patients. AB - A critical problem in leprosy is the relative deficiency of antigen-specific T cell-mediated immunity. We were successful in detecting a significant response to viable M. leprae in mononuclear cells isolated from the lymph nodes of lepromatous leprosy patients in contrast to the apparent M. leprae-specific energy seen in the peripheral blood. This observation suggests that antigen reactive lymphocytes are generated in the lymph nodes of lepromatous patients but the inability to detect them in the circulation may be due either to a different processing and presentation of mycobacterial antigens within the peripheral blood and lymph node compartments or to a selective sequestration of lymphocytes within the lymph node. PMID- 3045947 TI - Monoclonal antibodies against the major internal protein, p27, of a psoriasis associated retrovirus-like particle. AB - Four murine monoclonal antibodies (MoAb) were raised against p27 isolated from psoriatic scale. The MoAb recognized the antigen on a subfraction (0.1-1%) of psoriatic lymphocytes as well as in skin biopsies from psoriatic patients. In contrast, the antigen could not be detected on lymphocytes or in skin biopsies from healthy controls. A sandwich ELISA assay using the MoAb for detection of p27 is described. This method, together with a competition antibody-binding assay for distinction of epitopes, demonstrates that the MoAb recognize four different antigenic determinants on p27. PMID- 3045949 TI - An evaluation of the hemiplegic subject based on the Bobath approach. Part I: The model. AB - An evaluation, based on the Bobath approach to treatment has been developed. A model, substantiating this evaluation is presented. In this model, the three stages of motor recovery presented by Bobath have been extended to six, to better follow the progression of the patient. Six parameters have also been identified. These are the elements to be quantified so that the progress of the patient through the stages of motor recovery can be followed. Four of these parameters are borrowed from the Bobath approach, that is: postural reaction, muscle tone, reflex activity and active movement. Two have been added: sensorium and pain. An accompanying paper presents the evaluation protocol along with the operational definition of each of these parameters. PMID- 3045950 TI - [Nonspecific bronchial reactivity]. AB - Nonspecific bronchial reactivity is defined as the ability of bronchial smooth muscles to contract in response to non-allergenic (nonspecific) stimuli. Recent techniques for its measurement in different experimental and clinical situations have opened up new avenues in the understanding of asthma, airway infection, cough, and effects of pollution. The most recent data suggest that airway inflammation is the central mechanism of bronchial hyperreactivity, but the cells and mediators involved remain to be discovered. Bronchial reactivity as a test for clinical purposes needs standardization, careful establishment of normal values and assessment of validity in terms of the decision-making process. PMID- 3045951 TI - [Pleural diseases: an unusual cause of cough]. AB - The parietal pleura contains myelinated sensitive nerve fibers as well as mechanoreceptors which coordinate the breathing muscles. The small volume of fluid within the pleural space is in dynamic balance: hydrostatic and colloid osmotic pressures maintain a constant flow of fluid from the parietal to the visceral pleura, which turns into pleural effusion under pathological conditions. Separation of transudate from exudate is best done by calculation of the protein and LDH ratios in the pleural fluid and in serum. Transudates of cardiac origin are due to congestive left heart failure alone. In detecting subpulmonary pleural effusion, ultrasonography is more accurate than X-ray in lateral decubitus. Pleural exudate is investigated by chemical, microbiological and cytological examination of the fluid, complemented by pleural biopsy, fibre bronchoscopy and thoracoscopy. In this way less than 10% of pleural effusions remain unexplained. PMID- 3045953 TI - [Endocarditis or bacteremia caused by Streptococcus bovis and colorectal neoplasms]. AB - Streptococcus bovis is a relatively frequent causative agent of endocarditis or bacteriaemia, particularly in the elderly. In the past S. bovis has often been incompletely or even falsely classified. For therapeutic and prognostic reasons it is important to classify this agent exactly with biochemical methods even in the routine laboratory. Endocarditis or bacteriaemia due to S. bovis are often seen in conjunction with malignant, potentially malignant or benign colorectal neoplasias. After endocarditis or bacteriaemia due to S. bovis thorough investigation of colon and rectum is indicated. On the other hand, in presence of fever in patients with colorectal tumors, S. bovis bacteriaemia or endocarditis must be considered. The available literature is inconclusive on the question whether, after S. bovis endocarditis or bacteriaemia with initially normal colorectal findings, examination of the upper gastrointestinal tract and periodic inspection of the colon is needed. Up to now there has been no satisfactory explanation for the concomitant occurrence of endocarditis or bacteriaemia due to S. bovis and colorectal neoplasia. PMID- 3045952 TI - [Idiopathic lung fibrosis]. AB - Idiopathic pulmonary fibrosis (IPF), also called interstitial pneumonia or fibrosing alveolitis, is a progressive interstitial lung disease of unknown origin. Two distinct forms are known which differ in course, morphologic features and cytological findings in bronchoalveolar lavage: a cellular "desquamative interstitial pneumonia" and a hypocellular and more fibrotic "usual interstitial pneumonia". The two types appear to represent different stages of the same disease. The clinical findings in IPF are dyspnoea, unproductive cough, clubbing and rales. Pulmonary function tests reveal restriction, reduced diffusion capacity and hypoxaemia during exercise. Chest X-rays show localized linear or nodular densities usually accentuated at the lung bases; in 10% of patients with IPF, chest radiography is normal. To assess the inflammatory process bronchoalveolar lavage and transbronchial or open lung biopsy have proven necessary. Before beginning treatment the activity of the inflammatory process and degree of lung function impairment should be established. Therapy of IPF is unspecific and its aim is to suppress alveolitis and the fibrotic process. Corticosteroids are the initial therapy. If there is no improvement after two months, immunosuppressive agents are added. Regular clinical and functional follow-up is required. Course and prognosis vary in different forms of IPF. Without treatment, median survival in the cellular form is over 10 years and in the hypocellular form between 3.2 and 5.6 years. Only 12-30% of patients respond to antiinflammatory treatment. PMID- 3045954 TI - [Pharmacotherapy of cough]. AB - The optimal approach to the treatment of cough consists, first, in determination of its precise cause. Treatment should be directed at the specific etiology or the pathophysiologic mechanism responsible for cough. The outcome of specific treatment is almost always successful. However, there are also situations in which the cause of cough is unknown or in which the cough does not serve any useful purpose but prevents rest and sleep. In such cases symptomatic treatment with antitussives may be indicated. Centrally acting antitussives such as opium alkaloids, dextromethorphan and noscapine are the most effective drugs. The indication for expectorants lacks scientifically based support. PMID- 3045956 TI - [Compound strength of metal frameworks and adhesive agents in adhesive bridge technology. 1. A review of the literature and enamel etching]. PMID- 3045955 TI - [Cytological studies in pneumonological diagnosis]. AB - The following principles for the use of cytological examinations in pneumology are suggested: 1. The aim of cytological sputum examinations is diagnosis of tumors and not early detection of bronchial cancer in risk groups. 2. The diagnostic accuracy of cytological examinations depends mainly on the preparatory techniques. 3. Cytology and histology are complementary methods in the diagnosis of tumors and should always be combined. 4. Immunocytochemistry and electronmicroscopy are rarely indicated and are at best complementary to conventional microscopic diagnosis. 5. Cytological examinations for Pneumocystis carinii and other opportunistic germs in immuno-deficient patients should be confined to clinical centers with adequate experience. PMID- 3045957 TI - [Advances in research on the trichothecenes T-2, DON and NIV]. PMID- 3045958 TI - [Advances in research on targetting drug delivery of immunoliposomes]. PMID- 3045959 TI - [Theophylline and transplantation]. PMID- 3045961 TI - [Research on the genetic polymorphism of hydroxy metabolism of debrisoquine and its significance]. PMID- 3045962 TI - [The gene and function of apoproteins]. PMID- 3045960 TI - [Effects of the immune system on the endocrine system]. PMID- 3045963 TI - [3 leukocyte membrane glycoproteins related to adhesive function--LFA-1, Mac-1 and P150/95]. PMID- 3045964 TI - [A novel series of lipoxins, biologically active compounds formed from arachidonic acid]. PMID- 3045966 TI - Trapping with optical tweezers. PMID- 3045965 TI - [Endothelial cell-mediated regulation of microvascular permeability]. PMID- 3045967 TI - Time-resolved photoacoustic calorimetry: probing the energetics and dynamics of fast chemical and biochemical reactions. AB - Time-resolved photoacoustic calorimetry is a new experimental technique that measures the dynamics of enthalpy changes on the time scale of nanoseconds to microseconds for reactions initiated by absorption of light. When the reaction is carried out in water, it is also possible to obtain the dynamics of the corresponding volume changes. This method has been applied to a variety of biochemical, organic, and organometallic reactions. PMID- 3045968 TI - A novel instrument for separating large DNA molecules with pulsed homogeneous electric fields. AB - A new instrument has been developed for the electrophoretic separation of large DNA molecules that can independently regulate the voltage of each of 24 electrodes and allow the magnitude, orientation, homogeneity, and duration of the electric field to be precisely controlled. Each parameter can be varied at any time during the electrophoretic process. Thus distinct sets of conditions can be combined to optimize the separation of various fragment sizes in a single run. Independent control of electrode voltage allows all of the fields to be generated with electrodes arranged in a closed contour, independent of a particular geometry. This device increases both the resolution in any size range and the speed of separation, especially for DNA molecules larger than 3 megabases. PMID- 3045969 TI - Computational neuroscience. AB - The ultimate aim of computational neuroscience is to explain how electrical and chemical signals are used in the brain to represent and process information. This goal is not new, but much has changed in the last decade. More is known now about the brain because of advances in neuroscience, more computing power is available for performing realistic simulations of neural systems, and new insights are available from the study of simplifying models of large networks of neurons. Brain models are being used to connect the microscopic level accessible by molecular and cellular techniques with the systems level accessible by the study of behavior. PMID- 3045970 TI - Rearrangement of the bacterial chromosome: forbidden inversions. AB - The order of genes in the chromosome of enteric bacteria has been evolutionarily conserved despite the existence of mechanisms for rearrangement. Homologous chromosomal sequences in the same orientation recombine to form deletions or duplications. When homologous sequences in inverse orientation recombine, one expects to form an inversion of the intervening chromosomal segment. This expectation was tested by placing pairs of homologous sequences in inverse order at various points in the chromosome. Sequences at many pairs of sites (permissive) do recombine to generate the expected inversion, while the same sequences placed at other pairs of sites (nonpermissive) do not form an inversion. For the one nonpermissive interval tested, the missing inversion type can be constructed by an alternative transductional method; strains with this inversion are viable. Thus mechanistic limitations must prevent sequences at particular sites from undergoing the recombination event required to form an inversion. PMID- 3045972 TI - Anechoic abdominal mass on ultrasound. PMID- 3045971 TI - Site-specific integration of H-ras in transformed rat embryo cells. AB - A karyotypic analysis was performed on seven independently derived clones of primary rat embryo cells transformed by the ras oncogene plus the cooperating oncogene myc. The transfected oncogenes were sometimes present in amplified copy number, with heterogeneity in the levels of amplification. Some chromosomal features, such as aberrantly banding regions and double-minute chromosomes, typical of cells carrying amplified genes, were also seen in three of the seven cell lines. Underlying this heterogeneity there was an unexpected finding. All seven lines showed a common integration site for ras on the q arm of rat chromosome 3 (3q12), though some lines also had other sites of integration. In four of the lines integration of ras was accompanied by deletion of the p arm of chromosome 3 or its possible translocation to chromosome 12. PMID- 3045973 TI - Clinical aspects of abdominal masses in children. PMID- 3045974 TI - Adrenal neoplasms in children. PMID- 3045975 TI - Other abdominal and pelvic masses in children. PMID- 3045976 TI - [Epidemiological survey apropos of 7 Serratia marcescens infections]. PMID- 3045977 TI - [Fear and anguish in the works of Freud]. PMID- 3045978 TI - [Fear and madness]. PMID- 3045979 TI - [Dr. I.V. Cheshikhin, progressive Russian scientist]. PMID- 3045980 TI - [Insulin-induced lipodystrophy (insulin-induced panniculitis)]. PMID- 3045981 TI - [90th anniversary of the first Congress of the Russian Social Democratic Workers' Party]. PMID- 3045982 TI - [I. V. Davydovskii (1887-1987) (on the centenary of his birth)]. PMID- 3045984 TI - [Constipation]. PMID- 3045983 TI - [Purgative agents in the practice of today's physician]. PMID- 3045986 TI - [Diagnosis and prevention of complications from operations on the pancreas]. PMID- 3045987 TI - [Ultraviolet irradiation of the patient's own blood in suppurative-inflammatory diseases]. PMID- 3045985 TI - [Economical resections in lung cancer]. PMID- 3045988 TI - [Psychosocial factors and disease]. PMID- 3045989 TI - [Marxist-Leninist philosophy and biomedical publications]. PMID- 3045990 TI - [Electrocoagulation of the gallbladder bed following cholecystectomy with drainage of the abdominal cavity by the Spasokukotskii method]. PMID- 3045991 TI - [Community participation in the primary health care programs in the Third World]. PMID- 3045992 TI - [Bases for the study of the interaction of family, social networks and health service utilization]. PMID- 3045993 TI - [Notes on a disease in the village of La Magdalena. By F. Hinojosa, 1865]. PMID- 3045996 TI - Dipstick screening for urinary tract infection. AB - In screening for urinary tract infection the leucocyte esterase test will detect almost all samples with significant pyuria and bacteriuria, but is relatively nonspecific. The nitrite test is more specific but less sensitive and about one third of the urinary tract infections in a large group of children were missed. The combination of screening tests results in greater overall accuracy both in the diagnosis and exclusion of urinary tract infection. Almost all cases of urinary tract infection were detected when either the leucocyte esterase or the nitrite screening test or both were positive. If both tests are negative, urinary tract infection is virtually excluded and unless the child is symptomatic, further urinalysis is unnecessary. Laboratory urinalysis is, however, necessary if any one screening test for leucocyte esterase or nitrite (or protein or haemoglobin) is positive. Combined biochemical screening for urinary tract infection with dipstick test strips is reliable and allows early diagnosis and management. By avoiding unnecessary urinalysis it is cost-effective for the patient and will significantly reduce the laboratory workload. PMID- 3045995 TI - Malignant disease of the gastro-intestinal tract in blacks. Incidence and frequency distribution by age, sex and site at Ga-Rankuwa Hospital, Pretoria, 1976-1986. AB - The pathology records of Ga-Rankuwa Hospital for the years 1976 - 1986 were studied in respect of the numbers of gastro-intestinal malignant diseases diagnosed. Some tumours were seen more frequently in recent years, i.e. carcinoma of the oesophagus, stomach, colon and rectum. Whether this increased occurrence represents a true increase in this type of malignant disease is not proven. In comparison with other hospital reports, carcinoma of the oesophagus is very common and is certainly showing no decrease. PMID- 3045997 TI - Victoria Cottage Hospital--from the community, for the community. AB - Victoria Hospital, Wynberg, will have been in use for 100 years on 25 June 1988, serving the community of the southern suburbs of Cape Town as a general specialist hospital. When it opened its doors for the first time it consisted of two six-bed wards and one two-bed ward, the latter for paying patients. PMID- 3045994 TI - [Our inheritance from the early Vienna Medical School]. PMID- 3045998 TI - [The essence of prevention and the prophylactic orientation in health protection in a socialist state]. PMID- 3045999 TI - Diagnostic evaluation of extremity vascular injuries. AB - Diagnostic evaluation of patients with possible vascular injuries presents the physician with a number of circumstantial impediments that may limit the applicability of clinical signs and symptoms, noninvasive testing, and angiography. The challenge is to know these limitations and learn to work around them while still applying clinical skills, noninvasive tests, and angiography when appropriate and feasible and to the limits of their worth, rather than to resort to either routine angiography or blind exploration in every case. PMID- 3046000 TI - Thoracic great vessel injury. AB - Thoracic great vessel injury may be secondary to blunt, penetrating, blast, or iatrogenic trauma. A surgeon should be the initial evaluator of and decision maker for these patients, and the aortogram remains the gold standard for specific diagnosis of the arterial injuries except in those patients requiring emergency thoracotomy. Two general types of incisions are employed for these injuries: resuscitative and elective. PMID- 3046001 TI - Carotid and vertebral arterial injuries. AB - Injuries of the extracranial cerebral vessels represent only a small fraction of all reported arterial injuries but pose a significant dilemma over whether to repair or ligate the involved vessel. This article reviews recognition and repair of both penetrating and blunt injuries of the carotid and vertebral arteries, with special comment on the surgical exposure of the less accessible injuries. PMID- 3046002 TI - Upper-extremity vascular injuries. AB - Although upper-extremity injuries alone are usually not life-threatening, they can produce significant immediate or long-term morbidity, especially if there is an associated nerve injury. The diagnosis of an arterial injury may be readily apparent, but the excellent upper-extremity collateral circulation may create palpable distal pulses despite a significant proximal arterial injury. Therefore, a high index of suspicion and the liberal use of arteriography are necessary to avoid missing these injuries. Compression of the brachial plexus by a hematoma can produce a serious neurologic deficit. Prompt evacuation of the hematoma may significantly reduce the deficit, another fact that supports an aggressive surgical approach in these patients. The long-term results of upper-extremity vascular injuries are usually determined by the extent of any associated nerve injuries. PMID- 3046003 TI - Abdominal vascular injuries. AB - Abdominal vascular injuries remain rare in centers that primarily treat victims of blunt trauma, but when penetrating wounds of the abdomen are commonly treated, the incidence of abdominal vascular injuries is surprisingly high. With suitable management, many of these patients survive. PMID- 3046004 TI - Hemorrhage in major pelvic fractures. AB - Significant hemorrhage following major pelvic fractures should always be expected. Early recognition of such fractures during the resuscitation of any multiply injured patient is essential before instituting measures that might combat blood loss. In the majority of patients, simple resuscitative measures, including employment of the pneumatic antishock garment, will suffice. With certain types of fracture geography, the early application of external fixation devices may also play an important role. Increasingly popular has been the technique of diagnostic angiography and therapeutic embolization, applicable to approximately 3 per cent of all pelvic fracture patients. With exsanguinating hemorrhage, even the best equipped and most sophisticated major trauma centers can be taxed. The decision whether a patient should be taken directly to the operating room or to the angiography suite remains one of the most difficult for even the most highly skilled trauma surgeon. Patients with rapidly expanding or free rupture of pelvic hematomas noted at the time of celiotomy, or those with large open wounds, usually leave no recourse but to attempt direct operative control, to include even the most morbid option of a life-saving hemipelvectomy or corpectomy. More often, however, once other sources of surgically correctable hemorrhage are controlled or ruled out, diagnostic angiography followed by therapeutic embolization is a mainstay in the modern-day management of pelvic fracture hemorrhage. PMID- 3046005 TI - Hypothermia-induced coagulopathies in trauma. AB - Hemorrhage accounts for 90 per cent of deaths after abdominal injury, and half of these deaths are secondary to a recalcitrant coagulopathy. This review concentrates on our present knowledge of the role of hypothermia in trauma related coagulopathies and notes that preventing as well as treating these disorders remains the focus and the challenge of many investigators in the field of trauma. PMID- 3046006 TI - Popliteal and shank arterial injury. AB - Arterial injuries below the adductor hiatus in the lower extremity result in amputation more often than do injuries in any other site. The major deterrents to limb salvage are delay in diagnosis and revascularization and extensive adjacent bone and soft-tissue trauma. Rapid diagnosis and repair are needed for all popliteal artery injuries and for infrapopliteal injuries that compromise distal flow. The optimal management of asymptomatic injuries of single shank arteries with normal flow in uninjured parallel vessels is not defined. PMID- 3046007 TI - Management of venous trauma. AB - There has been considerable interest in the management of injured extremity veins since the American experience during the Vietnam War. Fortunately, there are an increasing number of reports from civilian experience in the United States that add valuable information. Although the controversy continues, it appears that there is merit in repair of many injured lower-extremity veins, particularly the popliteal vein when it is a single return conduit, assuming that the patient's general condition will permit, in an attempt to prevent acute venous hypertension initially and chronic venous hypertension subsequently. Figure 1 identifies the recovery potential that exists even if the initial venous repair fails. In contrast to thrombosis in the arterial system, recanalization is the rule in venous thrombosis. Patent valves can exist above and below the rather localized area of thrombosis. It appears that recanalization will prevent the problems of chronic venous insufficiency. It is obvious that many patients do well for years; however, the sequelae of acute venous hypertension may be more demonstrable after 10 or 15 years. There has not been similar evidence supporting a more aggressive approach in general in upper-extremity veins. However, it should be appreciated that a return pathway must remain patent, as noted in replantation of extremities. Obviously, there are differences in military and civilian wounds, with the former usually having more extensive soft-tissue destruction and obliteration of collateral veins and lymphatic channels. Unfortunately, many civilian gunshot wounds are being seen in the United States that are similar to the military type. We must not forget the lessons of the past, and we must continue to analyze our experience in the management of injured veins under a variety of conditions. PMID- 3046008 TI - Management of associated soft-tissue injury. AB - Initially, soft-tissue injuries associated with vascular trauma require management according to the general surgical principles of wound care. Antimicrobial prophylaxis should be instituted. The mechanism and extent of injury must be determined, and debridement and irrigation should be completed in the operating room with the intent to preserve vital structures, decrease bacterial contamination, and render the wound free of devitalized tissue. Prompt coverage of exposed vascular repairs is vital in order to minimize the chance for infection and to protect the repair from desiccation and subsequent trauma. Early primary closure of heavily contaminated wounds can have disastrous results. Therefore, questionable wounds should be managed by delayed primary closure after repeated irrigation and debridements, with the definitive decision as to the timing of wound closure being based on quantitative cultures. The plastic and reconstructive surgeon can aid the vascular surgeon in the primary or secondary closure of such wounds by bringing healthy, vascularized muscle to cover the vascular repair. Vascularized muscle has proved superior in the coverage and healing of contaminated wounds. If conditions permit, this can be most readily accomplished by transfer of local muscle. Musculocutaneous flaps are of special value in cases that require increased durability or where extended coverage is necessary. With either of these flap techniques, the surgeon must be thoroughly familiar with the anatomy, blood supply, and function of the local muscles. Free tissue transfer is to be utilized when local tissue is severely damaged in a way that precludes the use of adjacent muscle or myocutaneous flaps. Strict adherence to the principles and techniques of microsurgery, thorough evaluation of the zone of injury and recipient blood vessels, and understanding of the principles governing local muscle transfer are essential for a successful outcome. PMID- 3046009 TI - Nerve injury associated with acute vascular trauma. AB - Nerve injuries associated with acute vascular trauma are a formidable obstacle to restoring useful limb function. Nerve injury has become the primary cause of long term disability as the success rate of acute revascularization has risen to well over 90 per cent. Nerve injuries occur in roughly 60 to 70 per cent of upper extremity vascular injuries and in approximately 25 per cent of lower-extremity vascular injuries. These neural injuries account for a permanent disability rate of between 27 and 44 per cent. Establishing early continuity of completely transected nerves along with early exploration of lesions in continuity provides the best chance for timely axonal regrowth and reinnervation of the distal musculature. The use of intraoperative nerve action potential monitoring has allowed us to explore lesions in continuity earlier without unnecessary resection and suture of lesions showing sufficient axonal regeneration. At present, because of the relatively slow rate of axonal regeneration seen in axonotmetic or neurotmetic injuries (after repair), very proximal nerve injuries in either the upper or lower extremity will continue to lead to disappointingly poor motor recovery in the limb distally. PMID- 3046011 TI - Computer-assisted burn care. AB - The use of a computer does not take the thought process out of burn care. Rather, it provides constant user reinforcement while emphasizing key elements of burn care management. PMID- 3046010 TI - Vascular graft selection. AB - Twenty to thirty per cent of patients with arterial injuries and some patients with venous injuries require interpositional grafting. The first choice of grafting material for both arterial and venous injuries is autogenous vein. Injuries to large vessels such as the aorta and superior vena cava may necessitate synthetic prostheses. Synthetic aortic prostheses have excellent long term patency rates, but the same materials are much less likely to remain patent in the vena cava. Panel or spiral grafts constructed from saphenous vein appear to be the best replacement for this vessel. Autogenous veins are present in different diameters ranging from a mean of 6.4 mm in the saphenous vein to a mean of 1.8 cm in the internal jugular vein. The thickest autogenous vein is the saphenous vein, and thus it is preferred for medium-sized and small arteries. The authors prefer the larger 7.5-mm cephalic vein for replacement of medium-sized veins. In the absence of suitable saphenous vein, the cephalic vein is also the choice for arterial interposition grafts. Although there are few reports of the use of arterial autografts in vascular trauma, the surgeon should be aware that autografts may be ideal for vascular injuries in children and for isolated injuries with severe contamination. Finally, the use of synthetic grafts in injuries where adequate tissue coverage is not possible may result in immediate limb salvage, but the incidence of limb loss in this situation will be extremely high. PMID- 3046012 TI - [Contraception and the desire for children. Women's memoirs]. PMID- 3046014 TI - The Gittes procedure as an improved simplification of current techniques for vesical neck suspensions. PMID- 3046015 TI - The letters and friendship of Carrel and Cushing. PMID- 3046013 TI - A review of percutaneous drainage in splenic abscess. AB - The availability of ultrasonography and computerized tomography has significantly improved diagnostic capability in the evaluation of splenic abscess. In addition, recent evidence has shown that percutaneous drainage of splenic abscess is a safe and efficacious approach to therapy and is indicated especially when patients are seriously ill, postoperative or when the risks of general anesthesia, surgical drainage or splenectomy are substantial. Adequacy of response to percutaneous drainage correlates positively with the presence of unilocular collections that have a discrete wall without internal septations. In contrast, multiloculated or complex pyogenic splenic abscesses should usually be treated using operative intervention. Discussion of important features of this illness, as well as a comprehensive review of reported instances and guidelines related to percutaneous drainage of splenic abscess, are presented herein. A team approach, which uses the experience of imaging and surgical personnel, is invaluable in therapy when a splenic abscess is encountered. PMID- 3046018 TI - MF 20 microvascular Doppler. PMID- 3046016 TI - Modified and complete neck dissection in the treatment of squamous cell carcinoma of the head and neck. AB - The relative merits and indications for complete or modified dissection of the neck are straightforward. Which operation is selected centers on the perception of the value of the preservation or the risk with the loss of one or more of three structures: the spinal accessory nerve, the internal jugular vein and the sternocleidomastoid muscle. If these were the only issues, the choice of operation would be easy. Unfortunately, some of the most contentious issues in treatment of metastasis to the neck have been linked with the concept of the modified neck dissection. Such issues as combined multimodality therapy, preoperative and postoperative radiation therapy or surgical treatment alone, prophylactic dissection and bilateral simultaneous dissection of the neck are rightly or wrongly tied in with the type of dissection of the neck. More information, such as the certainty of the real risk factors in the neck with metastatic disease, the real value, if any, of adjunctive combined therapy and basic information about the role of the nodes in the neck, is necessary before the debate on the modified versus the radical dissection of the neck will end. PMID- 3046017 TI - Evaluation of endoscopic ultrasonography for the diagnosis of submucosal tumors of the esophagus. AB - Endoscopic ultrasonography was carried out on 55 patients whose X-ray films or endoscopic examinations indicated the presence of a submucosal tumor. Endoscopic ultrasonography revealed 8 cases of extraluminal compression and 48 cases of submucosal tumors. Histological studies were performed on 29 cases with submucosal tumors. In 28 of the 29 cases (97%) the location of the tumor in the esophageal wall was correctly estimated ultrasonographically, and appropriate treatment was selected. Tumors ranging from 3 to 50 mm in diameter could be measured accurately. This method may be helpful in follow-up studies. Endoscopic ultrasonographic findings, such as characteristics of the tumor border and internal echoes, were studied to predict the histological diagnosis of the tumor. Leiomyoma, cyst, granular cell tumor, lipoma, and intraluminal metastasis of esophageal cancer were all found to have specific ultrasonographic findings indicating the histological nature of the tumor. PMID- 3046021 TI - Sir Wylie McKissock. PMID- 3046019 TI - Reappraisal of the clinical expression of mixed cryoglobulinemia. PMID- 3046022 TI - Evolution of contemporary instrumentation for computer-assisted stereotactic surgery. AB - This article discusses the evolution of our stereotactic system which evolved from the commercially available Todd-Wells stereotactic instrument. The Todd Wells frame was originally designed for radiographically based, functional neurosurgical procedures. We modified it for computed tomography compatibility and later devised localization systems for magnetic resonance imaging and digital angiography. Based on the limitations in the original design applying to our own set of requirements, including tumor stereotaxis, we totally redesigned the system around the arc-quadrant principle of the original Todd-Wells instrument. While this intermediate system was being used in our surgical practice, further limitations were noted and corrected in the system we presently use. The present stereotactic frame is completely interactive with an operating room computer system. PMID- 3046020 TI - Therapeutic approaches in mixed cryoglobulinemia. AB - In summary, one can say that, although EMCG is a perplexing and puzzling disease, and although there are more enigmas than solutions, we can establish a way of handling the disease, knowing how to divide it into benign or acute forms, and establishing a treatment attitude according to that knowledge. PP and its modifications, in combination with immunosuppressive agents, are the corner stones to the acute form of the disease complicated by vasculitis, nephritis, chronic hepatitis and CNS involvement. Looking ahead, a new and more specific mode of treatment could emerge from our and others observations that the IgMs in the mixed cryoproteins are antiidiotypes to the IgGs. By producing autoantiidiotypes it might be possible to block some of the immune complex formation and to interfere with the cold precipitation, which is the basic pathophysiological process of essential mixed cryoglobulinemia. PMID- 3046024 TI - Historical vignette. The 1964 presidential address before the Society of Neurological Surgeons at its annual meeting in Rochester, Minnesota, May-8-9. By Bronson S. Ray. PMID- 3046023 TI - Magnetic resonance imaging of transdural herniation of a cervical disk. AB - A case of transdural herniation of a cervical disk is presented. The diagnosis was most clearly established by magnetic resonance imaging. The literature is reviewed and possible mechanisms of transdural herniation are discussed. PMID- 3046025 TI - Refraction problems after refractive surgery. AB - While refractive surgery such as radial keratotomy, epikeratophakia, and corneal relaxing incisions offer many potential benefits to patients, they can also generate optical problems such as overcorrection or undercorrection of the prior refractive error and variable vision. The authors offer suggestions for avoiding problems through proper patient selection; candid preoperative communication with the patient about possible difficulties and limitations; understanding of the physiological changes that may occur; and optical techniques for alleviating postoperative vision problems. PMID- 3046026 TI - Functional hemianopsia: a historical perspective. AB - The controversial history of functional hemianopsia is reviewed from Briquet's 1859 monograph on 430 cases of hysteria, through the 19th century works of Charcot, Freud, and Janet, and the observations of Fox and Wilbrand and Saenger in the early 20th century. More recently, concepts of this entity have been clarified by modern binocular testing of visual fields. PMID- 3046027 TI - Cardiac transplantation without cardiopulmonary bypass: experimental model to study growth of the transplanted heart. AB - An experimental model using surface-induced (20 degrees C) deep hypothermia and total circulatory arrest instead of cardiopulmonary bypass was developed for the study of growth of the transplanted heart. Autotransplantation of the heart was performed in 42 young dogs weighing from 4.4 to 9.0 kg (mean, 6.9 kg). Time of ischemia ranged from 26.0 to 60 minutes (mean, 43.4 minutes). Return of satisfactory cardiac function occurred in all but one animal. An early high mortality rate was due primarily to pulmonary complications, but with modifications to the technique, long-term survival increased to 70%. Early deaths (5 days to 13 weeks) of five dogs during preliminary trials were due to pleural effusion (2), sepsis (1), endocarditis (1), and ascites (1). There have been 14 long-term survivors (range, 194 to 498 days; mean, 264 days). Long-term survivors appear well, are active, and show satisfactory growth. This experimental model eliminates the need for heparinization and reduces the potential for complications associated with cardiopulmonary bypass in the dog. It avoids cannulations that might impinge on anastomotic sites. This model appears to be suited for studying growth of the transplanted heart. PMID- 3046028 TI - Changing concepts of deep venous thrombosis of the upper extremity--report of a series and review of the literature. AB - Deep venous thrombosis (DVT) of the upper extremity has recently been recognized as being more common than previously reported (probably because of the increasingly frequent use of subclavian venous access). A retrospective review of patients in whom subclavian or axillary DVT had developed in the past 6 years (1980 to 1986) was conducted at the Akron General Medical Center. The major cause identified was related to subclavian venous catheterization, which accounted for 39% of all instances of subclavian and axillary DVT. Our results are correlated with a review of the literature. In our review of studies in which subclavian venous catheterizations were prospectively examined with use of objective means of diagnosis, we found that 28% of all subclavian catheterizations had venous thrombosis develop, often subclinically. This is not an innocuous disease, as suggested in the past; in our series 12% of upper-extremity DVT had pulmonary embolization (PE). In reviewing the recent literature, we found an average 12.4% incidence of PE, which often occurs during anticoagulation treatment. Diagnostic modalities are discussed and treatment regimens are reviewed along with an extensive literature review. PMID- 3046029 TI - [Caries: summary observations of applied knowledge in view of new knowledge]. PMID- 3046030 TI - Periodontal treatment--new attachment. A review of the literature. PMID- 3046031 TI - [Leiomyosarcoma in the oral cavity: a literature review]. PMID- 3046032 TI - [Ameloblastoma: a retrospective study and case presentation]. PMID- 3046033 TI - [Water-alginate technique. Is vacuum mixing better than mechanical mixing?]. PMID- 3046035 TI - Problems of nomenclature and classification in medical expert systems. PMID- 3046034 TI - The diagnostic process, the diagnosis and homeostasis. PMID- 3046036 TI - [Adjuvant therapy following breast-sparing surgery of breast carcinoma: priority to chemo- or radiotherapy?]. PMID- 3046037 TI - [Significance and principles of antiemetic therapy in cytostatic treatment]. PMID- 3046038 TI - [Safety problems in the handling of cytostatics]. PMID- 3046039 TI - [The SAKK Melanoma Program (Swiss Group for Cancer Research)]. PMID- 3046041 TI - A combined method for the preparation of washed human platelets using prostacyclin and gel filtration. PMID- 3046040 TI - [Trends and perspectives in current cancer therapy]. PMID- 3046042 TI - Subunit B of factor XIII is present in bovine platelets. AB - Distribution of B subunit of coagulation factor XIII(FXIII B) in bovine platelets was determined. Intracellular location of FXIII B in platelets was confirmed by fluorescent antibody technique. Positive staining of FXIII B was observed with Triton-permeabilized platelets but not with non-permeabilized platelets. Immunoblots of plasma and platelet lysate revealed that the antibody was specific for FXIII B and this antigen existed in the supernatant of sonication-solubilized platelets. The amount of FXIII B in platelets was determined to be 17.3 +/- 6.7 ng/10(8) platelets by radioimmunoassay. This is the first visual and quantitative demonstration of intracellular FXIII B. PMID- 3046044 TI - Local plasminogen activator and prostacyclin-like activity after partial venous occlusion--an experimental study in the dog. AB - After 90 min. of partial venous occlusion (40%) in dog's femoral vein plasminogen activator (t-PA) was decreased in the pre-occlusion segment and maintained in the post-occlusion segment. Prostacyclin-like activity (PLA) was maintained in the pre-, but increased in the post-occlusion segment. Treatment with heparin (loading dose-70/kg; IV perfusion-1 U/kg/min) did not affect pre-treatment pattern of t-PA or PLA in either segments. Treatment with aspirin (IV perfusion-2 mg/kg/min) promoted profound depression of PLA in both segments while t-PA remained unchanged. Treatment with lidocain (loading dose-1.5 mg/kg; IV perfusion 0.2 mg/kg/min) did not affect either PLA or t-PA in the pre-occlusion segment, but a decrease of these two activities was observed in the post-occlusion segment. We conclude that, since thrombin, prostaglandins and white blood cells were not responsible for all changes observed at t-PA and PLA vessel wall levels, it is justified, to evaluate the role of rheological factors as the probable determinant of these changes. PMID- 3046043 TI - Plasma albumin is essential for collagen-induced platelet aggregation. AB - We found that platelets must have albumin on the surface to respond to collagen and aggregate. Albumin, however, was not absolutely necessary for ADP-, platelet activating factor-, serotonin- or thrombin-induced aggregation, while fibrinogen was required for ADP- or serotonin-induced aggregation. Immunofluorescent microscopy revealed that albumin was retained on gel-filtrated platelets but not on washed platelets. Albumin was not required for platelet adhesion to immobilized collagen. Without albumin thromboxane formation upon collagen stimulation was diminished. These data suggest that albumin is essential in some step(s) that results in production of thromboxane A2. PMID- 3046045 TI - [Non-pharmacological treatment of mild to moderate hypertension. A randomized, controlled study--results 1 1/2 years later]. PMID- 3046046 TI - [Cultivation of beta-hemolytic streptococcus from pharyngeal swabs. An evaluation of a method in general practice]. PMID- 3046047 TI - [Adrenal hematoma in newborn infants]. PMID- 3046048 TI - [Endoscopic ultrasonography of the upper gastrointestinal system]. PMID- 3046050 TI - History of lasers in medicine. AB - While the history of laser begins in 1951, the first medical application is reported by Goldman in 1962. In cardiovascular surgery McGuff first used a Ruby Laser in 1963 for the experimental ablation of atherosclerotic plaques. After a long time of investigations and new developments in laser technology first clinical applications were performed by Choy and Ginsburg in 1983. Since that time the effectiveness of laser angioplasty in coronary and peripheral vessel is investigated in several clinical trials and first results are encouraging, so that laser is about to find its place in the treatment of cardiovascular diseases too. PMID- 3046049 TI - [Somatotropin: structure, (bio)synthesis and species specificity]. AB - The literature on biosynthesis and species specificity of bovine somatotropin (BST) is reviewed. Somatotropin is a hypophyseal hormone consisting of 190 amino acids, having, among other things, a stimulating effect on milk synthesis. Recombinant DNA-techniques have been used since 1982 to produce BST and this has paved the way for large-scale use of r-BST. It is assumed that the somatotropin molecule has several active sites responsible for a multitude of biological effects following interaction with specific receptors. Somatotropin of different animals have a varying amino acid composition. Sixty-five per cent of human and bovine somatotropins are homologues in amino acid sequence. BST is found to be virtually inactive when used therapeutically in human individuals. PMID- 3046051 TI - Fundamentals of laser light interaction with human tissue, especially in the cardiovascular system. AB - The absorption of single photons in the molecules of biological tissue can induce various reactions. For the most medical laser applications the transformation from radiation energy into heat is relevant. The laser beam is used for coagulation or vaporization of tissue. The changes in tissue, which are created by light of different wavelengths depends on the thermal and optical properties (absorption and scatting) of tissue but also on the parameters of irradiation. As an example measurements from human skin are discussed. In the cardiovascular system laser light must have a clearly defined effect. Atherosclerotic plaques of different consistence have to be vaporized without damage of the vessel walls. From different reasons the usual medical CW-lasers, Argon-laser, CO2-laser and Nd:YAG-laser, are not optimal for direct ablation of arterial occlusions. In order to mimize reocclusion the walls of the channels have to be completely smooth and free of coagulation necrosis. This can be obtained by short laser pulses. Selection of a light wavelength, which is stronger absorbed in atherosclerotic plaques than in vessel walls and additional selective staining are two ways to reduce the risk of damaging the vessel walls. PMID- 3046052 TI - Laser application in bronchology. AB - The use of laser in bronchology plays a major role in the therapy of benign and malignant tumours. By flexible delivery systems, lasers can be used in combination with flexible bronchoscopes. By this benign tumours, especially in young children, can be ablated successfully without major surgical intervention. In malignant tumours laser ablation can only be used as a palliative procedure in cases untreatable by conventional surgery or prior to radiation or chemotherapy, in which the symptoms of hemopthysis or significant obstruction are indications for an urgent treatment. Especially in elder patients a quick relief of symptoms can be reached avoiding the high risks of an operation. A new and experimental field of laser application in bronchology is the tracheal reconstruction with laser assistance. In 10 Beagle dogs laser assisted tracheal anastomoses revealed good functional results with an uneventful postoperative course. Histology showed a normal healing and a good reconstruction of the ciliated endothelium without metaplastic changes. So this new technique may in future become an additional tool in the reconstructive surgery of the trachea and main bronchi. PMID- 3046053 TI - [Anterior gastropexy for gastroesophageal reflux in retarded children]. AB - With respect to the high incidence (10-15%) of gastro-esophageal reflux in mentally retarded children and the poor results of conservative treatment in this group, serious considerations should be given to antireflux surgery. In this study, the results of anterior gastropexy were analysed for a group of 17 retarded children, and compared with data from the literature about Nissen's fundoplication. With similar success rates, anterior gastropexy appears to have a negligible mortality rate. PMID- 3046054 TI - [Spontaneous remission of congenital liver cysts]. AB - A six-weeks old infant presented with vomiting, malnutrition, pneumonia and signs of biliary obstruction. The liver was enlarged; ultrasonography showed three large cysts in the right lobe. The cysts were thought to be congenital. Before we could differentiate them from other causes they disappeared spontaneously. To our knowledge, spontaneous regression of congenital cysts of the liver has not been reported previously. PMID- 3046056 TI - [Somatomedins and other growth factors]. PMID- 3046055 TI - [Medium-chain acyl-CoA dehydrogenase (MCAD) deficiency in 2 patients with symptoms of Reye syndrome]. AB - Two patients are described, who were submitted to our clinic with signs of the Reye syndrome. In both cases a medium-chain acylcoenzyme A dehydrogenase (MCAD) deficiency was diagnosed. This is an inborn error of the mitochondrial beta oxidation of fatty acids. Stimulation of the fatty acid oxidation in case of this enzyme deficiency might result in a metabolic crisis presenting clinically as the Reye syndrome. The structure of a fatty acid molecule and the process of beta oxidation of fatty acids are discussed shortly in this article. The most important clinical, diagnostic and therapeutic aspects of MCAD deficiency are presented next. A MCAD deficiency seems not to be rare in cases presenting as a Reye-like syndrome. Accurate distinction between MCAD deficiency and Reye syndrome can be made by gas chromatographic together with mass spectrometric analysis of urine. Investigation of so called crisis urine is of utmost importance. Confirmation of the diagnosis needs measuring of MCAD enzyme activity in cultured fibroblasts or in leucocytes of the patient. PMID- 3046057 TI - [Growth hormone treatment in secondary growth retardation]. PMID- 3046058 TI - Mutagenicity of the ash of rice straws by Ames' test. AB - Mutagenicity of fly ashes and bottom ashes of rice straw and rice husk was assayed by Ames' test. With respect to rice-straw ash, the extract from the fly ash was found to be more mutagenic than that from the bottom ash. In the case of rice husk, the mutagenicity of extract from the bottom ash was stronger than that from the fly ash. The extract from rice-husk bottom ash showed the strongest mutagenic activity among the four. PMID- 3046059 TI - Residual beta-cell function and metabolic stability in insulin-dependent diabetes mellitus. AB - Using the modified sensitive method of measurement of urinary C-peptide immunoreactivity (CPR) excretion, we studied whether positive correlation between residual beta-cell function and metabolic consequence is present in insulin dependent diabetes mellitus (IDDM) or not. After urinary CPR excretions were measured for 3 days in 40 IDDM, they were divided to three groups: group A, urinary CPR excretion less than 1 microgram/day (n = 16); group B, 1-4 micrograms/day (n = 15); group C, greater than or equal to 4 micrograms/day (n = 9). All subjects were treated with intensive insulin therapy for 1-2 months, and 1) fasting blood glucose (FBS) for one week, 2) 24-hr urinary sugar excretion (US) for one week, 3) M-value for daily variation of blood glucose, and 4) hemoglobin A1 (HbA1) were measured before and after intensive insulin treatment. And, we calculated mean and standard deviation (S.D.) of FBS and US. After intensive insulin therapy, a significant difference of the S.D. of FBS was seen between group A and group B (p less than 0.01). And, a significant difference of M-value was found between group A and group B (p less than 0.05). As significant differences were observed between group A and group B, which could not be divided into two groups by conventional CPR measurement, it seems likely that metabolic stability in IDDM mainly result from minimal remaining residual beta-cell function. PMID- 3046060 TI - 1988 Achievement Award Jeanne M. Manson. PMID- 3046061 TI - [Neutrophil elastase and alpha 1-proteinase inhibitor in the gingival fluid of patients with inflammatory diseases of the periodontium]. PMID- 3046062 TI - [Enzymatic fibrinolysis as limited proteolysis in the blood and mixed saliva of patients with lichen ruber planus of the oral mucosa]. PMID- 3046063 TI - [Enzyme therapy of patients with sialosis]. PMID- 3046064 TI - [Effectiveness of an improved method for the functional modelling of the base of a complete removable denture]. PMID- 3046066 TI - [Comparative laboratory evaluation of silicone impression materials used in preparing metalloceramic dentures]. PMID- 3046067 TI - [The history of dental congresses]. PMID- 3046065 TI - [Restoration of the occlusal height in the repeated prosthesis of the edentulous jaws by duplicating the occlusal surface]. PMID- 3046068 TI - [Mikhail Boleslavovich Iankovskii (1866-1923) (on the 120th anniversary of his birth)]. PMID- 3046069 TI - [Current aspects of antiviral and immune therapy of herpetic stomatitis]. PMID- 3046070 TI - [The influence of antral foreign body in the mucous membrane of Highmore's antrum]. PMID- 3046071 TI - Evolution and testing of the lacunar hypothesis. PMID- 3046072 TI - Spontaneous internal carotid artery dissection presenting as hypoglossal nerve palsy. AB - A 42-year-old man presented with right temporal headache, dysarthria, and dysphagia. On examination, he had a right hypoglossal nerve palsy. The diagnosis of right internal carotid artery dissection was suggested by magnetic resonance imaging and confirmed by carotid angiography. A dynamic computed tomogram demonstrated enlargement of the carotid artery. In carotid dissection, the hypoglossal nerve may be compromised by local factors as it passes close to the carotid artery in the neck. PMID- 3046075 TI - Multicenter trial of hemodilution in acute ischemic stroke. PMID- 3046074 TI - Thrombolytic therapy in cerebrovascular disease. PMID- 3046073 TI - Reversible cerebral segmental vasoconstriction. AB - Vasoconstriction is not recognized as a cause of cerebrovascular disease except in the vasospasm seen following subarachnoid hemorrhage and possibly in migraine. However, we found four patients to have transient, fully reversible vasoconstriction and dilatation prominently involving arteries around the circle of Willis. All four patients were evaluated for severe headaches and fluctuating or recurring motor or sensory deficits. No cause for the clinical syndromes and angiographic abnormalities was found. Similar patients are reported in the literature under various nosologies. This newly recognized clinical-angiographic syndrome should be differentiated from other known causes of vessel constriction and dilatation; the precipitants of reversible vasoconstriction may then be better defined. PMID- 3046076 TI - [Regulation of DNA synthesis in Acetabularia]. PMID- 3046077 TI - [Isolation and characteristics of the extracellular matrix of cultured cells]. AB - Sodium deoxycholate extraction was used to isolate extracellular matrix from various cultured cells: human and murine embryonic fibroblasts, epithelial lines of mouse (MPTR), rat (IAR 2 and IAR 20), pig (SPEV) and cow (FBT). Protein composition of the matrix was studied by sodium dodecyl sulfate polyacrylamide gel electrophoresis and immunofluorescence. The matrix morphology was investigated by scanning electron microscopy. In cell lines FBT and MPTR the major component of the matrix was laminin, whereas in other lines and fibroblasts it was fibronectin. The matrix of the majority of lines had a fibrillar structure, and the fibrils usually formed networks. MPTR cells had a punctate matrix composed of laminin and collagen type IV, densely covering the substratum. The treatment of the matrix by hyaluronidase and/or DNAase I did not influence its protein composition. The isolated matrix of different structure and composition may serve a biological substratum in studies of normal tumor cell behavior in tissue culture. PMID- 3046078 TI - [Mechanisms of the interaction of the leukemic cell with endogenous regulators of the proliferation of hematopoietic tissue]. AB - A substance inhibiting DNA synthesis in mouse leukemic cells was isolated from the regenerating calf spleen. When added to a suspension of leukaemic cells, this substance is adsorbed on their surface. The following changes in cell features being noticed: 1) a minute decrease in electrophoretic cell motility, 2) a decrease in esterase activity of the cells, 3) an increase in microviscosity of membrane lipids, 4) an increase in the intracellular pH values. With a longer contact with this substance, changes in nuclear chromatin structure were noticed, with special reference to weakened bonds between DNA and proteins. The data obtained are of significance for revealing molecular mechanisms of hematopoietic mediator action on target cells. PMID- 3046080 TI - [The coefficient of the relative position of particles and its use in the quantitative analysis of substance distribution in cells]. AB - A new method of objective estimation of material distribution in micro objects is described, namely, computing of the coefficient of mutual location of particles, while the cell image is presented as a digital map. The two methods of estimation of distribution of the material are compared - the coefficient of mutual location and the texture coefficient. Advisability of the parallel use of both the methods is substantiated. PMID- 3046079 TI - [Localization of neutral Mn-dependent DNAse in rat hepatocytes. An immunofluorescent study]. AB - Rabbit antibodies against the Mn-dependent DNAse were obtained. The specificity of the anti-DNAse antibody was established by the enzyme inhibition in vitro. The enzyme activity was inhibited by more than 75 and 54% using rabbit antisera and affinity-purified IgG. Localization of the enzyme in the rat hepatocyte nuclei was studied by indirect immunofluorescence. PMID- 3046081 TI - [Formation of an actin cytoskeleton and adhesive structures during the spreading of cultured cells]. AB - Fibroblast spreading was studied using immunofluorescent method that provided visualization of actin structures and adhesion contacts in the same cell. Four stages of actin system formation were observed. 1. Actin concentration in ruffles at the cell periphery. Formation of numerous dot-like contacts along the whole perimeter of the cell. 2. Formation of a circumferential actin bundle. Focal contacts are located at the outer edge of the bundle. 3. Gradual transformation of the circumferential bundle into actin network with triangular meshes. Peripheral (rather than internal) filaments of the network are associated with the focal contacts. 4. Appearance of the system of long straight actin bundles (stress fibers) associated with dash-like focal contacts. The stress fibers are supposed to arise from the triangular actin network which in its turn arises from the circumferential bundle. It is suggested that the formation of actin cytoskeleton is a process driven by the development of tensions in actin structures attached to the focal contacts at the cell periphery. PMID- 3046082 TI - [Switch in the synthesis from IgM to IgG in the human lymphoblastoid cell line RPMI-6410t as affected by human sera]. AB - The switch from IgM to IgG in lymphoblastoid cell line RPMI-6410t was induced by human sera. The factor inducing the switch was found in the human placental serum and in the serum of peripheral blood of healthy donors. The switch investigated is induced both in the initial line 6410t and in some IgM+ sublines derived from it. With the help of the cloning method some IgG+ sublines were developed with different IgG-synthesis levels from 6410t line and its IgM+ sublines after inducing the switch in them. Earlier another type of the switch induction from IgM to IgA was observed in the same line and its IgM+-sublines by the factors contained in some batches of fetal calf serum (FCSG+). Thus, the homogeneous IgM+ cell population is shown to be able to pass in vitro though two different stages of differentiation inherent to B lymphocytes in vivo. PMID- 3046083 TI - [A simple highly sensitive recording microspectrophotometer]. AB - A design of the recording microspectrophotometer is described. The instrument possesses an absolutely flat base line and quantum-noise limited detection threshold. Two principal elements of the design are the "jumping" stage, and the logarithmic amplifier with the phase-sensitive detector which converts the photomultiplier output into the optical density signal. The performance of the instrument is illustrated by the recordings of visual pigment spectra in single photoreceptors. PMID- 3046084 TI - [Identification and properties of insulin receptors in neuroblastoma C 1300 N 18 cells in various functional states]. AB - The presence of insulin receptors in neuroblastoma C 1300 N18 cells was shown by the method of the [125I]insulin binding with cells as well as by the electron cytochemical methods based on the analysis of the binding of colloid gold labelled insulin and of agglutinin of wheat embryos with the surface of plasma membrane. It is established that the number and distribution of insulin receptors depend on the functional state of cells. Expression of receptors on the membrane increases after 5'-deoxyuridine-induced differentiation of cells. Due to changes in the lipid composition of cells caused by the incubation with lecithin cholesterol liposomes (an increase in the content of cholesterol and its esters, as well as of unsaturated fatty acids and appearance of lysolecithin) the quantity of insulin receptors decreases and cell membranes are damaged. The lowering insulin regulation is not observed for insulin receptors of the neuroblastoma C 1300 N18 cells. PMID- 3046085 TI - [Fibrinolytic treatment in acute myocardial infarction. Results of randomized studies in recent years]. PMID- 3046086 TI - [Intravenous regional analgesia compared to infiltration analgesia in the reduction of distal forearm fractures]. PMID- 3046087 TI - [Acute urinary retention caused by incarcerated fibromyoma in the 8th week of pregnancy]. PMID- 3046088 TI - [Asymptomatic bacteriuria during pregnancy]. PMID- 3046089 TI - [Polycystic degeneration in the kidneys in dialysis patients]. PMID- 3046090 TI - Ultrasound in Medicine and Biology Prize (Francoise T. D'Astous and F. Stuart Foster). PMID- 3046091 TI - Antenatal fetal blood flow in the descending aorta and in the umbilical vein and their ratio in normal pregnancy. AB - One hundred and sixty sequential measurements of umbilical vein blood flow (UVBF) and descending aorta blood flow (DABF) in normal fetuses were performed in utero by the duplex real-time ultrasound and pulsed Doppler technique. Throughout pregnancy from 26 to 41 weeks the blood flow velocity of the umbilical vein (UV) and the descending aorta (DA) remained relatively constant, while the diameter of UV and DA, UVBF and DABF increased with advancing gestational age. Moreover, the UVBF-to-DABF ratio was nearly constant, with a mean value of 64% from 26 weeks onwards. The DABF-to-abdominal area (DABF-to-AA) ratio was constant throughout pregnancy, with a mean value of 7.0 ml/min/cm2. We call these two constants the umbilical-aortic index and descending-aortic index. These relationships may be of help in the diagnosis of intrauterine growth retardation and other fetal disorders. PMID- 3046092 TI - Use of amplitude-modulated focused ultrasound for diagnosis of hearing disorders. AB - Focused ultrasound with a frequency of 2.5 MHz and amplitude-modulated in the range of audiometric frequencies (125-8000 Hz) was employed to examine 50 normal hearing persons and in complex diagnostics of over 500 patients suffering from sensorineural deafness, otosclerosis, chronic otitis media, and acoustic neurinoma. Typical diagnostic signs of these diseases were revealed. Modulated ultrasound is shown to offer an improved diagnosis of hearing diseases. The results confirm the previous observation that amplitude-modulated ultrasound may excite not only the acoustic receptors but also the acoustic nerve fibres. Possible mechanisms of the stimulating effect of focused modulated ultrasound are discussed. PMID- 3046093 TI - Ultrasonic and histopathological correlations of deep focal hepatic lesions induced by stereotaxic Nd-YAG laser applications. AB - Thirty six deep focal hepatic lesions were induced in eleven piglets by means of an Nd-YAG laser. Laser shots of 80 W power and 10 s duration were used, the beam being transmitted through an echoguided stereotaxic handpiece. From day 0 to day 120, the animals underwent ultrasonographic and morphological controls. At lasering time an hyperechoic image--12-18 mm in diameter--appeared due to boiling of tissue water. During the twenty postoperative days the lesion core was an echo free area due to tissue vaporization, surrounded by an hyperechoic ring of increasing fibrosis, containing neovascularization and biliary ductules, while the hypoechoic outer area represented the peripheral halo of edema. In the long term, hyperechoic structures--swollen fibrotic septa of homogeneous fibrotic network--invaded the lesion site confirming good healing. PMID- 3046095 TI - Bibliography of biomedical ultrasound. No. 74. PMID- 3046096 TI - Correction of stress incontinence in women by simple sling operation. AB - A simple sling operation for the treatment of stress incontinence is presented. In entails the creation of a sling with the pubovesicocervical fascia as a base that is lifted upward to suspend the vesical neck by means of two 0 mersilene sutures introduced through two small colpotomies and guided into the retropubic area with a ligature carrier needle. The mersilene threads are knotted over the rectus fascia after making a small suprapubic incision. The procedure has the advantages of a small suprapubic incision, absence of large vaginal scars, and shortened hospital stay. PMID- 3046094 TI - A fundamental criticism of hydrophone-in-water exposure measurement. PMID- 3046097 TI - Alternative method of nerve-sparing when performing radical retropubic prostatectomy. AB - An alternative method of preserving the pelvic parasympathetic nerve plexus and potency when performing a radical retropubic prostatectomy is described. The prostate is removed in an antegrade manner as opposed to the classic retrograde approach with a current potency rate of 60 percent. The advantages of this technique are ease of defining the lateral vascular pedicles and late transection of the dorsal vein complex to minimize bleeding early in the procedure. PMID- 3046098 TI - Nosocomial urinary tract infections. AB - Hospital-acquired urinary tract infections, which account for approximately 40 percent of all nosocomial infections, often result in serious complications and ultimately lead to rising hospital costs. To combat the high incidence of nosocomial urinary tract infections, surveillance and control programs must be developed and carefully maintained by hospitals. When urinary tract infections cannot be prevented, empiric therapy with broad-spectrum antibiotics effective against beta-lactamase-producing bacteria should begin immediately. PMID- 3046099 TI - Further clinical experience with CO2 laser in microsurgical vasovasostomy. AB - Herein are reported the results obtained in 14 patients with the performance of vasovasostomy by carbon dioxide (CO2) laser. Fusion coagulation of the vas wall was successfully accomplished as demonstrated by postoperative sperm counts of over 20 million/mL in 86 per cent of the patients, and a pregnancy rate of 43 per cent in the group of patients operated on within less than ten years of original vasectomy. In contrast, those patients undergoing vasovasostomy ten years after original vasectomy had sperm counts of over 20 million/mL in 43 per cent of the cases, with a zero pregnancy rate. A significant reduction in total operative time was achieved as compared to the conventional microsurgical suture technique, corroborating the ability of the CO2 laser to simplify this technique while producing a sperm-tight anastomosis. One of the drawbacks of this operation is that it is not suited for the performance of a vasoepididymostomy which could be required in those cases in which sperm is absent from the vas fluid at the time of vasovasostomy. PMID- 3046100 TI - Management of children with hypertension from reflux or obstructive nephropathy. AB - During a ten-year period, 35 children presenting with vesicoureteral reflux, ureteropelvic junction obstruction, or a "small kidney" were found to be hypertensive. Of these, 15 subsequently underwent surgical procedures for relief of hypertension. Seven were "cured," six were "improved," and two were "unchanged." The severity of hypertension could not be correlated with the degree of reflux nor with the degree of obstructive uropathy. However, all children with reflux in our study who were hypertensive had some degree of calicectasis noted preoperatively on intravenous pyelogram. Also it was noted that hypertension may occur several years after successful anti-reflux surgery. Children with vesicoureteral reflux, ureteropelvic junction obstruction, or a small kidney need to have blood pressure determinations at regular intervals, even if all previous readings had been in the normotensive range and whether or not they were followed up medically or post surgically. We suggest that blood pressure determinations be made every three months for the first year after diagnosis of reflux or ureteropelvic junction obstruction, and at least once a year thereafter. PMID- 3046101 TI - Experimental Escherichia coli epididymitis in rabbits. AB - We describe an experimental model of bacterial epididymitis in New Zealand white rabbits. Inoculation of 10(7) colony-forming units of Escherichia coli in a retrograde fashion into the vas deferens reliably produced clinical, bacteriologic, and pathologic epididymitis. Inflammation was maximum at two weeks and subsided by one month without treatment. E. coli could be reisolated from the epididymides for up to two weeks post inoculation. We detected loss of spermatogenesis in both the ipsilateral and contralateral testes and the appearance of antisperm antibodies subsequent to the infection in some animals. There were 2 cases (11%) of histologic bilateral epididymitis after unilateral inoculation; one of these had bilateral clinical epididymitis with E. coli recovered from both epididymides at two weeks. PMID- 3046102 TI - Humoral hypercalcemia due to squamous cell carcinoma of renal pelvis. AB - We describe an unusual and rare case of humoral hypercalcemia due to Stage D squamous cell carcinoma of the renal pelvis in a patient with no evidence of bony metastases. The literature on humorally mediated hypercalcemia associated with epithelial tumors of the renal pelvis is reviewed. PMID- 3046103 TI - Bilateral cysts of tunica albuginea of testes. AB - A case is presented of a patient with bilateral cysts of the tunica albuginea. An orchiectomy had been accomplished on the right side, with confirmation of cysts of the tunica albuginea. Cysts of the contralateral testicle were diagnosed by testicular ultrasound examination. This case represents the first reported instance of bilateral cysts of the tunica albuginea. PMID- 3046104 TI - Primary carcinoma of seminal vesicle. Diagnosis assisted by sonography. AB - A case of primary carcinoma of the seminal vesicle in a nineteen-year-old man is described. This patient represents the twelfth case reported in the literature in Japan. The experience gained in this case indicated the potential usefulness of ultrasonography in the diagnosis of primary carcinoma of the seminal vesicle. PMID- 3046105 TI - Estrogens and phenylpropanolamine in combination for stress urinary incontinence in postmenopausal women. AB - Thirty-six postmenopausal women with objectively verified stress incontinence were treated with oral estriol (Triovex, 2 mg x 1) and phenylpropanolamine (Kontexin, 50 mg x 2) alone and in combination. After an initial four-week single blind period with phenylpropanolamine (PPA), either estriol or estriol and PPA were given randomly in four-week periods, in a crossover design. PPA and estriol in combination as well as PPA alone, raised the intraurethral pressure and significantly reduced the urinary loss by 35 per cent in a standardized physical strain test. In women with an initial low urethral pressure estriol also induced pressure increase. The leakage episodes and the assessed leakage amounts were significantly reduced by both estriol and PPA given separately as single treatment (28%) or when given as combined therapy (40%). Most of the women preferred the combined treatment to either drug alone. Additive but no synergistic effects are indicated. PMID- 3046106 TI - University of Edinburgh veterinary expeditions: 1966 to 1986. AB - Since 1966, teams of new graduates from the Royal (Dick) School of Veterinary Studies have organised eight veterinary research expeditions to Africa, tropical America and the Seychelles. The expeditions are now a regular feature of the school's activities. It is now possible to look back, learn and advise on future expeditions regarding timing, research and funding. PMID- 3046107 TI - Detection of antibodies against African swine fever virus using infected monolayers and monoclonal antibodies. AB - Three monoclonal antibodies, specific for porcine IgG, IgM and IgA, were used to develop isotype-specific immunoperoxidase monolayer assays for the detection of antibodies against African swine fever virus. A mixture of anti-IgM and anti-IgG monoclonal antibodies was used in an assay designed for screening sera. This test was compared with a commercially available ELISA by using experimental sera and field sera obtained after an outbreak of African swine fever on two farms in the Netherlands in 1986. Although the ELISA was less sensitive than the immunoperoxidase monolayer assay on sera taken early after infection, the tests were equally useful for screening purposes. The isotype-specific assays gave epizootiological information about the stage of infection on the two farms. PMID- 3046108 TI - Serological diagnosis of the porcine proliferative enteropathies: implications for aetiology and epidemiology. AB - Campylobacter mucosalis and C hyointestinalis have been associated with the proliferative enteropathies of pigs. An examination of the antibody response to these organisms and to the intracellular campylobacter-like organism was undertaken. Antibody to the campylobacter-like organism was predominantly IgM, short lived, and could be detected by an immunofluorescence test using bacteria released from lesions as antigen. The majority (75 per cent) of pigs with proliferative enteropathy at necropsy were antibody positive and a small number (4 per cent) of pigs in which lesions were not observed were found to have antibody. Antibody appeared to be correlated with the presence of lesions rather than with exposure to infection and was independent of the presence of antibody to C mucosalis or C hyointestinalis. In natural outbreaks of the disease antibody to the campylobacter-like organism was more prevalent than clinical signs in the affected animals. PMID- 3046109 TI - A major outbreak of botulism in cattle being fed ensiled poultry litter. AB - Eighty of a group of 150 housed beef cattle showed classical signs of botulism after eating a batch of ensiled poultry litter. Sixty-eight of the animals died and Clostridium botulinum type C toxin was detected in 18 of 22 sera examined. C botulinum organisms were isolated from the ensiled litter and type C toxin was demonstrated in samples of decomposed poultry carcases present in the litter. This outbreak of bovine botulism was the most serious to have been recorded in Europe and was the first associated with feeding ensiled poultry litter. PMID- 3046110 TI - The late J.A. Bogan. PMID- 3046111 TI - Sampling scheme. PMID- 3046113 TI - Dr J. T. Edwards and breeding cattle for the tropics. PMID- 3046112 TI - Efficacy of piperazine dihydrochloride against Ascaris suum and Oesophagostomum species in naturally infected pigs. AB - A controlled trial was performed to evaluate the efficacy of piperazine dihydrochloride in a new granular formulation (Ascarex D) against naturally occurring infections with Ascaris suum, Oesophagostomum dentatum and O quadrispinulatum. Treatment effects were estimated on the basis of parasites recoverable from the intestinal contents. Given orally at 200 mg per kg body weight the compound showed an efficacy of 99 to 100 per cent against A suum and the nodular worms. Egg excretion of the respective species was reduced by 98 per cent and 100 per cent six days after treatment. No adverse reactions were observed after the treatment. PMID- 3046114 TI - Pathophysiological effects of endotoxins in ruminants. 1. Changes in body temperature and reticulo-rumen motility, and the effect of repeated administration. AB - Data from the literature on the clinical effects of bacterial endotoxins in ruminants are reviewed. Special attention is paid to the effects on body temperature and reticulo-rumen motility. Furthermore, the effects of repeated intravenous injection of endotoxin are summarised. Pathophysiological disturbances after intramammary infusion of endotoxins proved to be identical to those found after intravenous injection of non-lethal doses. Strikingly, however, no marked inhibitory effect on rumen motility nor abortion was observed after intramammary infusion of endotoxins. Moreover, in cows that were made tolerant to endotoxin by daily intravenous injections, intramammary infusion of one-fifth of this daily dose produced a maximum effect on body temperature and plasma Zn concentrations. This suggests that inflammatory endogenous mediators were released in the udder and then absorbed into the blood circulation, rather than the absorption of endotoxin. PMID- 3046115 TI - Pathophysiological effects of endotoxins in ruminants. 2. Metabolic aspects. AB - Metabolic disturbances following intravenous and intramammary administration of endotoxins in ruminants are described. In contrast to the similarity in response of blood biochemical parameters after intravenous and intramammary administrations of endotoxins, responses in plasma concentrations of enzyme activities, the thyroid hormones, cortisol, and somatotropin differ markedly. Biochemical changes in blood after endotoxin administration are predominantly dose-dependent; thus some of the biochemical parameters - especially plasma concentrations of Fe and Zn - serve also to evaluate the effects of certain drugs in endotoxin models. Changes in milk composition have been documented only after intramammary infusion of endotoxins and can partly be explained by the increased permeability of the blood/milk barrier. Appearance and production of milk returns to normal within a week after intramammary endotoxin treatment, indicating that the mammary gland is only temporarily damaged by endotoxin-induced mastitis. PMID- 3046116 TI - Bovine respiratory syncytial virus-specific monoclonal antibodies. AB - Five hybridomas were produced which secreted monoclonal antibodies to bovine respiratory syncytial virus (BRSV). Two antibodies (8G12, 15C7) neutralized the virus and inhibited syncytia formation in vitro. These monoclonal antibodies also stained, by indirect fluorescent assay, an external envelope protein of living virus-infected cells, and recognized the 48k subunit of the viral fusion protein by Western blot analysis of bovine respiratory syncytial virus-infected cell lysates. Three other monoclonals (6A12, 14D3, 14E3) stained, by indirect fluorescent assay, acetone-fixed virus-infected cells but not living cells. Three hybridomas (6A12, 8G12, 15C7) secreted monoclonal antibody of isotype IgG1, k; two hybridomas secreted monoclonal antibody of isotype IgG2a, k. This apparently is the first report of monoclonal antibodies specific for BRSV glycoproteins. PMID- 3046117 TI - [The genesis of a complex discipline--molecular biology--within the movement of ideas in the first half of the 20th century]. PMID- 3046118 TI - A carboxyl-terminal peptide of the DNA-binding protein ICP8 of herpes simplex virus contains a single-stranded DNA-binding site. AB - The DNA-binding protein ICP8 of herpes simplex virus is a multifunctional protein which is required for viral replication. To identify the single-stranded DNA binding domain of the protein, recombinant plasmids containing the 5' or 3' coding portion of the ICP8 gene or the intact gene were constructed and transcribed using SP6 RNA polymerase. The resulting RNA was translated in vitro to produce a 62,000-Da amino-terminal peptide, a 69,000-Da carboxyl-terminal peptide, or the intact protein. When these were analyzed by single-stranded DNA cellulose column chromatography, large amounts of the intact ICP8 bound to the columns while small amounts of the carboxyl-terminal peptide and undetectable amounts of the amino-terminal peptide bound. The majority of the carboxyl terminal peptide which bound eluted from the columns with the same salt concentration as the intact ICP8. The in vitro synthesized intact protein had the same affinity for single-stranded DNA-cellulose as ICP8 purified from infected cells. These results suggest that the carboxyl-terminal portion of ICP8 contains a single-stranded DNA-binding site. PMID- 3046119 TI - Rift Valley fever virus M segment: cellular localization of M segment-encoded proteins. AB - The Phlebovirus Rift Valley fever virus (RVFV), like other members of the Bunyaviridae family, matures intracellularly at the smooth-surfaced vesicles in the Golgi region of infected cells. Here we show that in cultured cells the RVFV glycoproteins G2 and G1 accumulate and are retained at this site. To investigate the parameters governing this subcellular localization, we have engineered portions of the cloned RVFV M segment (which encodes a 14- and a 78-kDa protein, in addition to glycoproteins G2 and G1) into vaccinia virus. Immunofluorescent analysis of cells infected with a vaccinia virus recombinant containing the entire open reading frame of the RVFV M segment revealed Golgi localization for glycoproteins G2, G1, the 78-kDa protein, and Golgi as well as some reticular distribution for the 14-kDa protein. These distributions paralleled those seen in authentic RVFV-infected cells. RVFV-vaccinia virus recombinants possessing progressive deletions within the 152 amino acid preglycoprotein sequence of the M segment were analyzed for possible effects on the cellular distribution of G2 and G1. Removal of the first 130 amino acids of the open reading frame amino-terminal to the mature glycoprotein coding sequences, while abolishing production of the 78- and 14-kDa proteins, did not alter the Golgi location of G2 and G1. The data suggest that Golgi-specific signals reside within the G2 and/or G1 glycoprotein sequences. The use of vaccinia virus recombinants provides a genetically manipulable expression system with which to further investigate the sequences involved in the intracellular localization of these Phlebovirus proteins. PMID- 3046120 TI - Resistance of the 64K protein of budded Autographa californica nuclear polyhedrosis virus to functional inactivation by proteolysis. AB - The 64K surface protein of budded Autographa californica nuclear polyhedrosis virus (AcMNPV BV) is known to play a role in the functional entry of AcMNPV BV into Spodoptera frugiperda IPLB-SF-21 cells by adsorptive endocytosis. AcV1, a neutralizing monoclonal antibody, reacts with the 64K protein and in doing so prevents efficient entry. In this communication we report that treatment of AcMNPV BV with either trypsin or proteinase K cleaves the 64K protein into one major fragment of 34.6K and two minor fragments of 36K to 37.2K that are retained with the virus. All of the fragments are glycosylated. Protease treatment does not reduce viral infectivity, but it does result in the destruction of the AcV1 reactive epitope; thus AcV1 is not able to neutralize protease-treated AcMNPV BV. Polyclonal antiserum to BV is able to recognize both cleaved and uncleaved 64K and neutralize both protease-treated and untreated virus. Protease treatment does not diminish the sensitivity of AcMNPV BV to chloroquine, but it does cause the virus to become more susceptible to inactivation by 2-mercaptoethanol (2-ME) even though exposure to 2-ME does not result in dissociation of the fragments from the virus. PMID- 3046122 TI - [Detection of thrombocyte antibodies using the ELISA technic. II. Comparison of thrombocyte reactivity on the ELISA and immunofluorescence test]. PMID- 3046121 TI - In vitro assembly of the outer shell of bacteriophage phi 6 nucleocapsid. AB - Following dissociation of bacteriophage phi 6 nucleocapsid (NC) by EDTA, a particle composed of protein P8 and corresponding to the outer shell of the NC was assembled in vitro in the presence of Ca2+ and Mg2+. Assembly was obtained from soluble protein constituents above 100 micrograms/ml and was optimal within a temperature range of 22-30 degrees. Assembly did not require the presence of genomic RNA. Crosslinking results of intact NCs and in vitro-assembled outer shells suggested that protein P8 dimers are the structural subunits of the shell. Analysis of the assembly kinetics by electron microscopy suggested that ring-like particles of uniform size, packed in flat hexagonal arrays, are intermediates in outer shell assembly. PMID- 3046123 TI - [The effect of a single administration of captopril on portal hemodynamic values]. PMID- 3046124 TI - [Recent findings in infectious endocarditis]. PMID- 3046125 TI - [Use of the computer in the diagnosis of congenital heart defects]. PMID- 3046126 TI - [C-peptide and glucose tolerance in thyrotoxicosis]. PMID- 3046127 TI - [Ultrasonic diagnosis of gunshot wounds of the extremities (experimental research)]. PMID- 3046128 TI - [Experience in organizing research on donor blood for the presence of antibodies to the human immunodeficiency virus]. PMID- 3046130 TI - [Induction casting of dentures in a dental polyclinic]. PMID- 3046129 TI - [Renin and aldosterone levels of the blood in young people during adaptation to military service]. PMID- 3046131 TI - [History of military medical expertise in the Navy]. PMID- 3046132 TI - [40th anniversary of the World Health Organization]. PMID- 3046135 TI - [Determination of blood flow volume by Doppler echocardiography]. PMID- 3046134 TI - [Further study of the isoelectric characteristics of the tick-borne encephalitis virus]. PMID- 3046133 TI - [The biochemistry of the African swine fever virus]. PMID- 3046136 TI - [Venous urography from the viewpoint of the nephrologist]. PMID- 3046137 TI - [Campylobacter pyloridis and diseases of the stomach and duodenum]. PMID- 3046138 TI - [Lipoproteins in diabetes mellitus]. PMID- 3046139 TI - [Non-digitalis cardiac inotropic compounds for parenteral and oral use]. PMID- 3046140 TI - [Bacterial infections of the lung]. PMID- 3046141 TI - [Incidence, diagnosis, prevention and treatment of insulin allergy]. AB - In the Scientific Institute for Endocrinology, Geriatrics and Gerontology, Sofia, 1723 diabetic patients were treated with insulin during the last 3 years. In 150 of them insulin allergy was supposed. The patients sensitivity toward the various insulin preparations was determined by intradermal test. Insulin allergy was found in 29 patients (1.68%). Insulin allergy was found more frequently in patients using acid insulin preparations and by frequent changes of the type of preparation. It is recommended that in special categories of diabetic patients- such as growing up, pregnant women, with complications of insulin treatment--only neutral, swine and purified insulin preparation should be used. PMID- 3046142 TI - [Syncopes and orthostatic hypotension]. PMID- 3046144 TI - [Outpatient echographic follow-up of patients with Fraley's syndrome]. AB - The follow-up of the degree and evolution of the upper hydrocalicosis and of the parenchymatous changes in patients with Fraley's syndrome is very important. All authors agree that these patients should be followed up periodically. 20 patients with Fraley's syndrome were followed up and the authors conclude that the echographic examination is a very reliable method for prophylactic medical examinations since it is informative, harmless and economically advantageous. PMID- 3046143 TI - [The membrane theory of the etiology and pathogenesis of hypertension]. PMID- 3046145 TI - [Threatened and actual extension (recurrence) of acute myocardial infarct]. PMID- 3046146 TI - [An acute recurrence of focal-segmental glomerulosclerosis in a kidney transplanted from mother to son with a rapid decline in kidney function]. AB - A case is presented of a 19-year-old man suffering from focal-segmental glomerulosclerosis with terminal chronic renal failure to whom a kidney taken from his mother was transplanted. There was high blood-group and tissue compatibility between mother and son. The initial result was good, the transplanted kidney functioned well-diuresis of 3300 ml with high proteinuria. Gradually the diuresis fell to 100-200 ml. From the 29th day following the transplantation pulse urbason therapy was applied for 3 days but without effect. This led to the resumption of hemodialysis and removal of the transplanted kidney. The microscopic examination of the kidney revealed massive focal segmental glomerulosclerosis which had led to terminal chronic renal failure. The rapid severe relapse of the disease in the transplanted kidney is explained with the malignancy of the disease and the very high compatibility between donor and recipient. It is recommended that renal transplantation in patients with focal segmental glomerulosclerosis should not be performed with a kidney taken from a parent. PMID- 3046147 TI - [The use of metamizole as a risk factor in the occurrence of agranulocytosis]. AB - The risk of agranulocytosis following the use of metamizole was studied by the method of controlled cases. All cases of agranulocytosis and their control cases in Sofia for the period 1982-1986 were included in the study. The mean agranulocytosis morbidity for this period was 3.16 cases for one million population for one year. The etiologic fraction was 7.1%. The relative risk of agranulocytosis following the use of metamizole is 1.25, the maximal risk is 0.03 cases for one million persons using metamizole for one year and the maximal lethal risk is 0.007 cases for one million persons using metamizole for one year. The use of metamizole in Bulgaria for the same period was studied, too. The data presented as a number of defined daily doses used by 1000 population for one day show that the use of metamizole in this country is high. A tendency toward an increase of 7.3% per year was found during the investigation. At the same time the agranulocytosis morbidity remained constant during the period of investigation. The results of the study lead to the conclusion that the use of metamizole is not related to a higher risk of agranulocytosis. PMID- 3046148 TI - [Arteriosclerosis risk factors: new aspects]. PMID- 3046149 TI - [Mutagenic effect of mercury]. PMID- 3046150 TI - [Physicians in the Uprising of 1863]. PMID- 3046151 TI - [Contribution of Poles to European cardiological science (to the end of the 19th century)]. PMID- 3046152 TI - [Treatment of cholesterol cholelithiasis with chenodeoxycholic acid]. PMID- 3046153 TI - [The role of diet in carcinogenesis]. PMID- 3046155 TI - [Responsibilities of public health in sauna construction and operation]. PMID- 3046154 TI - [The Dermato-syphilological Clinic of the Stefan Batory University in Wilno headed by Prof. Tadeusz Karol Pawlas 1935-1938]. PMID- 3046156 TI - [New aspects on the etiology and pathogenesis of arteriosclerosis from the pathologic viewpoint]. AB - Three main atherogenic processes are recognised today: hyperlipemia, arterial wall injury and parietal thrombosis. The role of hyperlipemia is supported, among other things, by the following: 1. Experimentally, protracted hyperlipemia can reproduce faithfully the lesions and all complications of advanced human atherosclerosis. 2. Immunohistochemically, plaque lipoproteins are identical with certain blood lipoproteins. 3. The incidence of atherosclerosis in different populations roughly parallels the average blood lipid levels of these populations. 4. Dietary and pharmacological reductions of blood lipid levels in certain populations have reduced the clinical manifestations of atherosclerosis in these populations. Prolonged hyperlipemia generates arterial plaques by causing penetration of blood lipids into the myocytes of the inner arterial wall, immigration of lipid-laden monocytes into the subendothelial space, and increased endothelial permeability for blood lipoproteins and mitogens. All types of arterial wall injury diminish the endothelial barrier and increase endothelial permeability for blood lipoproteins and mitogenic factors. Seven groups of naturally occurring arterial insults are recognised today: hemodynamic turbulence, hypertension, metabolic insults (including hyperlipemia), immune insults, viruses, exogenous chemicals, and obstruction of adventitial lymphatics. These insults usually cause a functional increase of endothelial permeability (when mild) or a loosening of interendothelial junctions (when intense). Parietal thrombosis develops practically only in atherosclerotic-almost never in normal arteries. It is most frequently initiated by tiny breaks of plaque surfaces, breaks which expose blood to the highly thrombogenic collagen and lipid masses that abound in the atherosclerotic--but are absent from the normal arterial wall. parietal thrombi are overgrown by endothelium, turned into fibrous tissue and incorporated into the underlying plaques, whose thickness they can thus greatly increase. PMID- 3046157 TI - [Ambulatory physical therapy rehabilitation with interval walking-gymnastics therapy in patients with chronic peripheral arterial occlusive disease]. AB - A ambulatory physical therapeutic rehabilitation by means of interval-run gymnastic-therapy was examined by a prospective study of 43 patients with peripheral arterial disease of the legs and was estimated as practicable. Improvements of walk distance, lengthenings of the pain times in the standardized move up and down-test, decreases of the blood-pressure gradients by ultrasonic Doppler measurements, shortenings of the half-life periods of xenon-clearance, improvements of the metabolism of lactate, improvements of the parameter of metabolism of lipids and uric acid were proved. Successes of therapy were not proved by oscillography and venous occlusion plethysmography. The expense was registered by the loss of working time and discussed. PMID- 3046158 TI - [Ultrasonic diagnosis in aneurysm of the abdominal aorta]. AB - The sonographic diagnostics of an aneurysm of the abdominal aorta has obtained a high value on account of the high reliability and the rapid availability of the investigation in connection with the absolute safety of the method. Form, extension, calcification, haemodynamics, thrombosis and dissection of an aneurysm of the aorta can sonographically be demonstrated and measured by longitudinal and transversal sections. As non-invasive approach the ultrasound tomography in the order of the graduated diagnostic method stands before the computed tomography which is not always at once available and before the angiography which have their qualification in the case of a planned operative approach above all for the purpose of the exact topographic differentiation between suprarenal and infrarenal extension. PMID- 3046159 TI - [Russia-Hall relations in medicine of the 18th century. III. Halle physicians as research travelers and members of expeditions]. AB - In the history of Russian expeditions in the 18th century several physicians and natural scientists who were educated in Halle and temporarily working, respectively, at the university of the city on river Saale occupy a respectable position. Names such as Buxbaum, Messerschmid, Stoeller, Lerche and Pallas are mentioned with high respect up to the present time. The results of their investigations in Sibiria, in Caucasia and in Persia are regarded as pioneer achievements which became the prerequisite for the investigations of the 19th century. PMID- 3046160 TI - A short commemoration of Camillo Golgi 60 years after his death. PMID- 3046161 TI - Injuries of the hand in athletes. PMID- 3046162 TI - [Possible mechanisms of false positive reactions in immunoenzyme analysis]. PMID- 3046163 TI - [Use of a new antibacterial preparation, tomicide, in the treatment of patients with pustulous diseases of the skin]. PMID- 3046165 TI - Bunyavirus-vector interactions. AB - Recent advances in the genetics and molecular biology of bunyaviruses have been applied to understanding bunyavirus-vector interactions. Such approaches have revealed which virus gene and gene products are important in establishing infections in vectors and in transmission of viruses. However, much more information is required to understand the molecular mechanisms of persistent infections of vectors which are lifelong but apparently exert no untoward effect. In fact, it seems remarkable that LAC viral antigen can be detected in almost every cell in an ovarian follicle, yet no untoward effect on fecundity and no teratology is seen. Similarly the lifelong infection of the vector would seem to provide ample opportunity for bunyavirus evolution by genetic drift and, under the appropriate circumstances, by segment reassortment. The potential for bunyavirus evolution by segment reassortment in vectors certainly exists. For example the Group C viruses in a small forest in Brazil seem to constitute a gene pool, with the 6 viruses related alternately by HI/NT and CF reactions, which assay respectively M RNA and S RNA gene products (Casals and Whitman, 1960; Shope and Causey, 1962). Direct evidence for naturally occurring reassortant bunyaviruses has also been obtained. Oligonucleotide fingerprint analyses of field isolates of LAC virus and members of the Patois serogroup of bunyaviruses have demonstrated that reassortment does occur in nature (El Said et al., 1979; Klimas et al., 1981; Ushijima et al., 1981). Determination of the genotypic frequencies of viruses selected by the biological interactions of viruses and vectors after dual infection and segment reassortment is an important issue. Should a virus result that efficiently interacts with alternate vector species, the virus could be expressed in different circumstances with serious epidemiologic consequences. Dual infection of vectors with different viruses is not unlikely, because many bunyaviruses are sympatric in nature. For example, the Ae. trivittatus-cottontail rabbit and the Ae. triseriatus-squirrel arbovirus cycles are sympatric in the ecotone between their respective grassland and forest ecosystems (LeDuc, 1979). Should a LaCrosse virus variant or reassortant evolve that was efficiently vectored by Ae. trivittatus mosquitoes, significantly more human infections with La Crosse virus would likely occur. Unlike Ae. triseriatus, Ae. trivittatus mosquitoes are not restricted to forested areas and consequently are more likely to encounter and to feed upon humans.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3046164 TI - Structure, function, and intracellular processing of paramyxovirus membrane proteins. AB - Paramyxoviruses are a fascinating group of viruses with diverse hosts and disease manifestations. They are valuable systems for studying viral pathogenesis, molecular mechanisms of negative strand viral replication, and glycoprotein structure and function. In the past few years this group of viruses has received increased attention and as a result there is a wealth of new information. For example, most of the genes of many paramyxoviruses have been cloned and sequenced. The recent availability of sequence information from a number of paramyxoviruses now allows the direct comparison of the amino acid sequence and determinants of secondary structure of analogous genes across the family of viruses. Such comparisons are revealing for two reasons. First, results provide clues to the evolution of these viruses. Second, and more importantly, comparisons of analogous genes may point to sequences and structural determinants that are central to the function of the individual proteins. Below is a comparison of five of the paramyxovirus genes with a discussion of the implications of common structural determinants for function, intracellular processing, and evolutionary origin. The focus is on the paramyxovirus membrane proteins, although other proteins are discussed briefly. PMID- 3046166 TI - [Max Grob and pediatric surgery in Zurich--a memorial lecture]. PMID- 3046167 TI - Primary megaureter in the neonate with prenatal or postnatal diagnosis. AB - Twenty-nine cases of primary megaureter diagnosed before the age of 3 months are grouped in this diagnostic and therapeutic study. Seventy-five per cent were discovered by antenatal sonography due to the presence of pelvicalyceal dilatation without localising precisely the position of obstruction, twenty-five per cent were detected by the occurrence of early urinary tract infection. Four patients were operated during the first few months. 2 diversions and 2 reimplantations. Fifteen including the 2 with early diversion were submitted to later reimplantation at the average age of 15 months. 12 patients were treated conservatively by antibiotics and regular supervision. The respective results of the operated and non-operated patients were evaluated comparing the initial dilatation and the last IVU using the classification of Beurton. Regardless of the marked initial dilatation, significant spontaneous regression or complete resolution were often seen during the first year of follow-up. Thus surgery should be deferred initially even for the severely dilated forms, except in a few critical cases where a diversion is indicated. Reimplantation should be deferred for some months unless deterioration is noted, or performed only after one year or 18 months if regression is insufficient, thus optimising operative conditions. PMID- 3046169 TI - [Congenital hydrocephalus. A multicenter retrospective longitudinal study of newborn infants treated with shunts 1976-1985]. AB - A multicentric retrospective evaluation of 54 children was performed who were born between 1976 and 1985 with a clinically recognisable hydrocephalus and thereafter shunted. The 4 topics of the study included: 1) Analysis of pregnancy, antenatal diagnostic procedures and parturition, 2) Preoperative evaluation and intraoperative findings, 3) Long-term follow-up and 4) Prognosis (actual status) of the 45 surviving patients at the end of the study (May 1987). Patients with neonatal hydrocephalus were observed in 11% of all children shunted during the same time in the hospitals included in this study. During this period the incidence of new cases did not decrease. PMID- 3046168 TI - [The Lich-Gregoir extravesical antireflux-plasty]. AB - Between 1974 and 1986 96 children underwent the Lich-Gregoir antireflux procedure, in 63 patients on both sides in one session. 17 patients had double ureter. In totally 153 controlled operations we found 4 persistent reflux (2.6%) and 1 stenosis (0.7%). The success ratio is, therefore, 96.7%. PMID- 3046171 TI - Use of cultured keratinocytes in the treatment of severe burns. AB - A 6 1/2-year-old child was admitted to the emergency ward for third degree burn injuries representing 40% of body surface. Shock therapy was first applied. After debridement and in a series of operating sessions hands were grafted with full thickness skin and most other wounds were covered with mesh grafts. On the 14th day after admission a piece of approximately 5 cm2 split thickness scalp skin was used to expand the keratinocytes by cell culture techniques according to the method of Rheinwald and Green (7). After 20 days 4 sheets of cells of first subculture, each 10 cm in diameter, representing a surface of approximately 300 cm2, were implanted on the front of the left thigh, which was burnt third-degree deep. Light microscopy of punch biopsies from mesh grafted and keratinocyte implanted sites taken 5 months after grafting showed a well differentiated epidermis overlying scar tissue. The following conclusions were drawn: Autologous keratinocytes did take on a third degree wound. No basic difference was observed neither clinically nor histologically between mesh-grafted and keratinocyte implanted sites. The epidermis formed by keratinocyte implantation lacked pigmentation but presented an aesthetically better appearance than the mesh-graft treated sites. Given the scarcity of donor sites and the results obtained by implanting keratinocyte cultures, this latter technique may be resorted to in any extended and deep burn injury. PMID- 3046170 TI - [Congenital hydrocephalus--an analysis of the course of pregnancy, prenatal studies and labor]. AB - An analysis of the pool of our patients revealed few disturbances during pregnancy. In two cases the malformations were possibly due to drugs. Prenatal sonographic diagnostics were performed in one-third of the cases or documented: in 60% of the investigated cases a hydrocephalus was found, this being only 20% of the whole pool. The major part of the births were performed by Caesarean section, mainly because of a prolongation of the birth directly or indirectly due to the large circumference of the head. The hereditary trait of associated malformations and malformations of the central nervous system was frequent. Therefore, we advocate a better prenatal work-up and a birth by planned Caesarean section. PMID- 3046172 TI - [Barber epitaph tells dental history]. PMID- 3046173 TI - [Postage stamps honor pioneers in dentistry]. PMID- 3046175 TI - [Silanization of the adhesive bridge attachments]. PMID- 3046174 TI - [Numerical test of binding of metallo-ceramic bond]. PMID- 3046176 TI - [Scintigraphy in evaluation of healing of bone transplants]. PMID- 3046178 TI - [Construction of removable cast anterior partial prosthesis for the mandible]. PMID- 3046177 TI - [Significance of the DE-System for complete dentures]. PMID- 3046179 TI - [Indications and technology of a torsion bridge]. PMID- 3046180 TI - [Problems of prosthetic concern with unilateral distal extended partial dentures]. PMID- 3046181 TI - [Modified application of a precision joint between anchor elements and the basic framework. 2]. PMID- 3046182 TI - [Immunologic aspects of habitual abortion and intrauterine fetal death]. AB - Habitual abortions and intrauterine death represent diagnostic and therapeutic problems in spite of application of near all diagnostic possibilities. The appearance of autoantibodies to nuclear and extractable nuclear antigens as well as occurrence of phospholipid-antibodies may be associated with negative course of pregnancy. Furthermore problems of autoantibodies to DNA, the role of HLA system trophoblast antigens and other reasons will be explained. Treatment facilities are referred, too. PMID- 3046183 TI - [Evolution of the otolithic membrane: its structural organization]. PMID- 3046185 TI - [Methicillin-resistant Staphylococci]. PMID- 3046184 TI - [Evolution of the otolithic membrane: its functional organization]. PMID- 3046186 TI - [The history of plague control]. PMID- 3046187 TI - [Comparative analysis of the protein composition of the external membrane of a virulent strain of Shigella sonnei and of genetically related avirulent bacteria]. AB - The outer membrane proteins of intact S. sonnei cells of a strain virulent in the keratoconjunctival test contain up to 24 polypeptides; one of them (with a molecular weight of about 70 KD) is absent in similar preparations of avirulent nonpenetrating shigellae in phase I, genetically related to the above strain. Study of the composition of the outer membrane proteins of S. sonnei cells in phase II has revealed that these cells, when compared with smooth bacteria, show qualitative and quantitative differences in their polypeptide fractions; of these, three fractions are characteristic of rough bacteria. PMID- 3046188 TI - [Clinico-experimental study of a dried selective medium for isolating fungi in the genus Candida]. AB - The trial of a newly developed dried selective medium for the isolation of fungi of the genus Candida, involving the inoculation of 103 museum yeast strains and 542 specimens of pathological material, has been carried out. The data obtained in this trial indicate that the proposed medium has advantages over wort agar and Sabouraud medium with antibiotics in the germination index, in the ability to ensure fungal growth after the inoculation of pathological material, and in selective properties. The medium ensures the intactness of the morpho-cultural, biochemical and serological properties of the fungi. The results of the trial recommend the newly developed preparation for laboratory practice for the isolation of yeast-like fungi from clinical material in the diagnosis of candidiasis, intestinal microflora disturbances, as well as for isolation of the fungi from various environmental objects. PMID- 3046189 TI - [Use of enzyme-antibody immune complexes in serological analysis]. AB - In model experiments with simulation of rabbit immune response to mouse IgG and antibody production in echinococcosis patients the indirect enzyme-linked immunosorbent assay has been several times more sensitive with peroxidase antiperoxidase and catalase-anticatalase complexes than with enzyme-antiglobulin chemical conjugates. The immune complexes have been found to retain their activity for 6 months and longer. PMID- 3046190 TI - [The standardization of conjugates for the indirect solid-phase immunoenzyme analysis reaction]. AB - The comparative study of several batches of conjugates has revealed that enzyme immunoassay techniques can be used for the standardization of conjugates by their affinity level. The study has shown that this can be done only if the concentration of specific antibodies in the conjugate is known and the amount of the conjugate is in excess to that of the antigen adsorbed on the plate. PMID- 3046191 TI - [Further development of the Expanded Programme on Immunization of the World Health Organization]. PMID- 3046192 TI - [The role of viral and bacterial infectious processes in the etiology and pathogenesis of acute inflammatory diseases of the bronchi and lungs]. PMID- 3046193 TI - [Use of a genetically labelled strain for the analysis of Yersinia pseudotuberculosis population dynamics in natural soils]. AB - The Rifr mutant of Y. pseudotuberculosis, capable of producing pure cultures in media with a high content of rifampicin, has been used for an accurate quantitation of this microorganism in various kinds of natural (nonsterile) soil in controlled laboratory and field experiments. The main biological characteristics of the mutant have been identical to those of the parent strain. The first experiments have shown that the initially high concentration of Y. pseudotuberculosis in the soil gradually decreases in 2 months. The share of this microorganism in the natural microflora of the soil seems to be rather small, which probably explains the cause of low indices of spontaneous contamination of the soil in nature. PMID- 3046194 TI - [40th anniversary of the World Health Organization]. PMID- 3046195 TI - [Mechanism of the nonspecific protective action of Shigella antigens]. AB - The mechanism promoting the nonspecific action of antigens obtained from S. flexneri and S. sonnei by a sparing method has been studied. These antigens stimulate the T- and B- systems of immunity, that is followed by activation of myelopoiesis and the humoral protective factors of the body, which seems to underlie the formation of resistance to infection caused by nonspecific microorganisms. PMID- 3046196 TI - [Use of quantitative immunofluorescence for determining the adsorption capacity of magnetic immunosorbents]. AB - The adsorption capacity of microgranulated polyacrylamide magnetic immunosorbents has been studied by the method of quantitative immunofluorescence as applied to the causative agents of plague, cholera, and melioidosis. Similar regularity in the dynamics of antigen adsorption on the granules of magnetic immunoadsorbents has been established. This regularity consists in the direct relationship between this process and the concentration of infective agents interacting with magnetic sorbents. PMID- 3046198 TI - [Chemotaxis of Yersinia pseudotuberculosis as a mechanism in its search for target tissues of the host organism]. AB - Y. pseudotuberculosis cells grown at biologically low temperature have been shown capable of chemotaxis with respect to carbohydrates and amino acids. During cultivation at 36-37 degrees C Y. pseudotuberculosis cells retain this property for 10-15 hours and then lose it. The mechanism of chemotaxis makes it possible for Y. pseudotuberculosis to "find" human and animal tissues and can facilitate the realization of the pathogenicity potential of these bacteria. When administered orally to mice motile bacteria, i. e. those grown at 6-8 degrees C, have been more virulent for the animals than nonmotile ones cultivated at 36-37 degrees C. PMID- 3046197 TI - [The protective properties of myelopeptides in the development of infectious processes caused by bacteria of the genus Salmonella]. AB - The protective properties of myelopeptides in the development of bacterial infection in mice and young pigs, caused by S. typhimurium 415, S. cholerae-suis 1422 and 370, have been studied. Myelopeptides have been found to possess protective properties when injected into animals infected with S. typhimurium and S. cholerae-suis in lethal doses. The best protective effect (survival rate of 100%) has been achieved by the injection of myelopeptides 24 hours before challenge. Myelopeptides have also been found to promote the weight gain of young pigs infected with S. cholerae-suis. PMID- 3046199 TI - [Characteristics of the antigen-binding properties of lymphocytes in experimental Salmonella infection]. AB - The data on the count of antigen-binding lymphocytes in the blood and lymphoid organs of mice in the course of Salmonella infection are presented. The reaction used for their detection is specific. Antigen-binding lymphocytes detected in mice in Salmonella infection belong to T- and B-cell populations and carry surface receptors belonging to IgA and IgM. Study of antigen-binding lymphocytes 1-20 hours after infection may be helpful in the specific diagnosis of Salmonella infections. PMID- 3046200 TI - [Effect of intraperitoneal administration of Streptococcus group A and its cell fractions on the development of adjuvant arthritis in rats]. AB - The effect produced by the intraperitoneal injection of live and heat-killed group A streptococci, the fractions of their cell walls (both intact and sonicated) and cytoplasm was studied on 450 white rats with experimental adjuvant arthritis (AA). The injection of live streptococci into rats with AA decreased the swelling of joints (by 70-80% in the second half of the experiment), reduced the titers of rheumatoid-like factor (RLF), and inhibited the development of polyarthritis. The use of heat-killed streptococci gave a less pronounced antiarthritic effect, while the fraction of streptococcal cell walls, similarly to live streptococci, decreased the swelling of joints (by 27-64%); at the same time a considerable drop in the titers of RLF was observed in 3 experiments, and the development of polyarthritis was registered in 38% of the test animals and in 62% of the control animals. In rats with AA the cytoplasm not subjected to ultracentrifugal purification decreased the swelling of joints (by 21-50%) and the titers of RLF. In this case the development of polyarthritis was observed in 48% of the test animals and in 70% of the control animals. PMID- 3046201 TI - [The role of the Guberniya reform of 1775 in organizing psychiatric care in Russia]. PMID- 3046202 TI - [Depression and disorders of circadian rhythm (a review)]. PMID- 3046203 TI - [Humeroscapular periarthrosis in cervical osteochondrosis (a review)]. PMID- 3046204 TI - [Sources of the classifications and nosological systematics of psychiatry in the 18th century]. PMID- 3046205 TI - [Raynaud's phenomenon (a review)]. PMID- 3046206 TI - [Differences in the comprehension of neurotic disorders in slowly progressive schizophrenia (a review)]. PMID- 3046207 TI - [Information value of initial clinical signs for the prognosis of outcome in the first 24 hours after craniocerebral injury]. AB - The informativeness of clinical indicators for predicting lethal and favourable outcomes during the first 24 hours after a head trauma has been investigated. A pool of clinical findings about the status of 302 patients examined according to a uniform technique has been analyzed using a packet of the MEDSTAT-85 software. The authors present an optimal set of clinical signs for predicting fatal and favourable outcome within the first 24 hours after the trauma with an 83% probability rate. PMID- 3046208 TI - [Information value of the clinical, personality and psychosocial characteristics of schizophrenics for the prognosis of results of rehabilitative therapy]. AB - The authors compared formalized case records contained in the data bank "Rehabilitative Automated Information System" (RAIS) for two groups of schizophrenics who have received a course of restorative therapy. The first group consists of 173 patients with A and B remissions, the second one, of 155 patients with D and C remissions and without remissions. The authors have analyzed 487 morbid characteristics of patients and identified 209 of them, which allow the differentiation of the groups under study at a statistically significant level. Forty of the most informative signs were utilized for constructing on the basis of image-recognizing algorithms of determinants for the individual prognosis of the remission type. PMID- 3046209 TI - [Use of a point-scale assessment of patient status in the surgical treatment of craniocerebral injuries]. AB - The indications and contraindications for surgical treatment in craniocerebral trauma (CCT) were based on estimation of the patient's condition in marks; the dynamics of changes of the results of the estimation in the pre- and postoperative periods were studied. A total of 375 patients with CCT were examined in different medical institutions according to a unified method. Neurosurgical interventions were carried out on 155 patients. All patients who underwent operation when their condition was rated below 15 marks died on the immediate postoperative days, whatever their age and whatever the time of the operation after the trauma. The probability of a favourable outcome increased to 40% in a condition rated 21-30 marks on the day of the operation and reached 69% when it was above 30 marks. PMID- 3046210 TI - [Early surgical treatment of patients with ruptures of cerebral arterial aneurysms]. PMID- 3046211 TI - [The theory of dielectrolysis iontophoresis]. PMID- 3046212 TI - [Possibilities of replacing bone tissue]. PMID- 3046213 TI - [Etiopathogenesis of the congenital hip dislocation syndrome]. PMID- 3046214 TI - Testosterone and the control of hypothalamic GnRH. AB - The aim of the present study was to test the hypothesis that the restoring effect of testosterone on hypothalamic GnRH stores in orchidectomized rats might be linked to the intrahypothalamic transformation of the hormone into estrogenic metabolites. To this purpose, advantage has been taken of the availability of the potent antiestrogen, tamoxifen (TMX). Different groups of adult male rats castrated since 4 weeks were submitted to a 6-day treatment with testosterone propionate (TP, 2 mg/rat daily); TMX (50 or 200 micrograms/rat daily); or TP (2 mg/rat daily) plus TMX (50 or 200 micrograms/rat daily). The animals were sacrificed 24 h after the last injection, and hypothalamic GnRH content was measured by radioimmunoassay. The results have confirmed the ability of TP to counteract the decreasing effect of orchidectomy on hypothalamic GnRH stores, and have shown that TMX does not have any intrinsic activity on this parameter. Furthermore, TMX at either dose used in the present experiments, was found not to be able to abolish the restoring effect of TP on MBH-GnRH stores. It is concluded that the action of testosterone on hypothalamic GnRH does not require the conversion of the hormone into estrogens. PMID- 3046215 TI - 4-hour urinary and serum immunofluorimetric assay for luteinizing hormone to detect onset of preovulatory LH surge. AB - The initial surge of LH is an important preovulatory event. The validity of a time-resolved immunofluorimetric assay method for determining LH both on the samples of serum and urine are reported in our study. A good correlation exists between the serum and urinary LH with the presence of a shorter interval in the length of the LH increase in the urinary dosage in respect to the serum dosage. There emerges when comparing the RIA and IFMA techniques that a good correlation exists both in the urinary and in the serum samples. However the values found with the IFMA method are on the average inferior (ca. 50%) in respect to those obtained with the RIA technique. PMID- 3046217 TI - On being aware: patient recall of intraoperative events. PMID- 3046216 TI - GnRH agonists in the treatment of endometriosis. AB - GnRH agonists, synthetic peptide analogs of GnRH, desensitize pituitary receptors for the native molecule, thus causing reversible hypogonadotropic hypogonadism. Numerous clinical studies have suggested that these compounds are efficacious in the treatment of endometriosis, but it is not clear whether they are superior to the other drugs used in treatment of this disease. The frequency of recurrence of pain symptoms at the end of treatment is high and the data on recovery of fertility are conflicting. Long-term administration of GnRH agonists is a safe and well tolerated treatment but its role in the management of endometriosis is still not well defined. PMID- 3046218 TI - The process of implementing a statute: how it works. PMID- 3046219 TI - Comparison between fluctuating PEEP and conventional PEEP in dogs with lung injury induced by blood aspiration. AB - It has been documented that in some patients with acute hypoxic respiratory failure the application of positive end-expiratory pressure (PEEP) may produce no improvement or even a deterioration of pulmonary oxygenation due to an increase in ventilation-perfusion mismatching. Fluctuating PEEP (F-PEEP) is a newly developed PEEP in which end-expiratory pressure (EEP) is periodically changed within a certain range. In a dog model with localized lung injury induced by the aspiration of non-heparinized blood (2 ml.kg body weight-1), F-PEEP in which the EEP was periodically changed from 0.5 to 1.5 kPa at frequencies of 10 min, and conventional PEEP with 3 different fixed EEPs, 0.5, 1.0 and 1.5 kPa (C-PEEP0.5, C PEEP1.0 and C-PEEP1.5) were each applied for 60 min. F-PEEP produced a periodical change in PaO2 and hemodynamic variables including cardiac output, and in comparison with C-PEEP0.5, C-PEEP1.0 and C-PEEP1.5, a significantly greater improvement of A-aDO2 and dynamic compliance with relatively large cardiac output in the low EEP phase. These results suggest that F-PEEP is a useful mode of artificial ventilation for treating some kinds of acute hypoxic respiratory failure due to increased ventilation-perfusion mismatching. PMID- 3046220 TI - Fluctuating PEEP (F-PEEP) versus conventional PEEP in dogs with asymmetrical lung injury. AB - Fluctuating PEEP (F-PEEP) is a newly developed PEEP in which end-expiratory pressure (EEP) is periodically changed within a certain range. In a dog model with unilateral lung injury induced by the introduction of hydrochloric acid, F PEEP in which the EEP was periodically changed from 0.5 to 1.5 kPa at periods of 6 min, and conventional PEEP (C-PEEP) with an optimized EEP of 1.0 kPa, were each applied for 30 min. F-PEEP produced a significantly greater improvement of PaO2 and intrapulmonary shunt (QS/QT) than C-PEEP, and at the low EEP phase, the greatest improvement accompanied by an increased dynamic compliance and a large cardiac output was obtained. These results suggest that F-PEEP provides a useful mode of artificial ventilation for the treatment of unilateral lung injury. PMID- 3046222 TI - Fluctuating PEEP versus conventional PEEP in diffuse and unilateral lung injury induced by oleic acid. AB - Effects of fluctuating positive end-expiratory pressure (F-PEEP), in which end expiratory pressure (EEP) was periodically changed from 0.5 to 1.5 kPa with a periodic time of 6 min, and conventional PEEP (C-PEEP) with a fixed EEP of 1.0 kPa, were comparatively studied in diffuse (Group I) and unilaterally dominant lung injury (Group II). Although F-PEEP produced cyclic alterations of PaO2 in both groups, PaO2 changed in proportion to EEP in Group I and in reciprocal proportion to EEP in Group II. There was no significant difference between PaO2 and QS/QT during F-PEEP and those during C-PEEP in Group I, whereas in Group II, F-PEEP produced a significantly greater improvement of pulmonary oxygenation at the low EEP phase than C-PEEP. In both groups, the degree of hemodynamic depression was proportional to EEP. These results suggest that F-PEEP should be indicated for acute hypoxic respiratory failure with uneven distribution of lung injury. PMID- 3046221 TI - The cardiovascular effects of anticholinergic agents administered during halothane anaesthesia in children. AB - The cardiovascular effects of anticholinergic agents administered during halothane anaesthesia were studied in 31 children aged 1-12 years undergoing peripheral orthopaedic surgery. Either normal saline, glycopyrrolate (10 micrograms.kg-1) or atropine (20 micrograms.kg-1) was administered in randomized double-blind fashion during the induction of anaesthesia with halothane while the electrocardiogram was continuously recorded. After induction, the children were paralyzed with atracurium, intubated, and ventilated. Anaesthesia was maintained with N2O/O2 and halothane (up to 2.5% inspired). The concentrations of expired CO2 and halothane were measured continuously using mass spectrometry. Sixty-one percent (19/31) of the children developed one or more dysrhythmias. Junctional rhythm occurred in 74% (14/19) of the children with dysrhythmias, developed early during induction (mean +/- s.d. time = 2.29 +/- 2.0 min after commencement of induction), and usually resolved before the administration of the study drug (8/14). All dysrhythmias initially occurred before or during induction and none developed during intubation, during incision, during the maintenance of anaesthesia, or after the administration of anticholinergic agents. The data suggest that: 1) a combination of factors present during halothane induction is highly dysrhythmogenic especially for junctional rhythm; 2) junctional rhythm will resolve spontaneously; 3) the administration of an anticholinergic agent during halothane induction is safe but may be unnecessary in children greater than 1 year of age; and 4) the dysrhythmogenic factors present during induction are attenuated during the maintenance of halothane anaesthesia. PMID- 3046224 TI - Hemodynamic significance of internal carotid artery disease. AB - Neurologic symptoms in the region of an internal carotid artery stenosis are considered to be embolic in most instances. Only in a subgroup has carotid occlusive disease with impairment of the collateral supply, caused a state of hemodynamic failure with marked reduction of perfusion pressure. Though unproven, it is reasonable to assume that without surgical intervention, the risk is higher than average for patients with hemodynamic failure. Equally, should there be any postoperative improvement of cerebral blood flow or neurologic deficits, it should be looked for in this group. Thus, it is necessary to distinguish those with low perfusion pressure from the population of patients with carotid artery disease. Preoperative clinical evaluation and direct visualization of the carotid bifurcation should be supplemented by indirect physiological tests which allow assessment of collateral perfusion. Examination of periorbital flow direction or oculoplethysmography could be used as a screening procedure. Negative tests most certainly rule out any severe pressure gradient across the stenosis, irrespective of the luminal reduction. A positive result, on the other hand, should be further quantified since most indirect tests become positive at relatively small pressure gradients. Studies of cerebral blood flow at rest and during cerebral vasodilation makes it possible to identify patients with severe reduction of cerebral perfusion pressure. Such hemodynamic failure of one hemisphere may be identified in most cases by a conventional non-invasive xenon-133 technique and stationary detectors. Smaller focal regions of hypoperfusion may be identified by computer emission tomography, either by the detection of single-photon emission or by paired detection of annihilation photons. Endarterectomy does improve cerebral hemodynamics in terms of increased flow through the reconstructed vessel and elimination of pressure gradients. The cerebral blood flow, though remains unchanged in the majority of patients, at least when measured at baseline. Only in those patients with a reduction in perfusion pressure can a significant improvement in baseline flow occur. Flow reserve determined by cerebral vasodilation, however, will improve in most patients with hemodynamic failure. In addition, some patients in the low-pressure group develop marked, but temporary, hyperperfusion after reconstruction of very high grade carotid stenosis. This is considered a result of chronic low perfusion pressure with subsequent loss of autoregulation, and autoregulatory control is first regained after some days.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3046223 TI - Sensory and secretory potencies and differentiations of the central nervous system. AB - Secretory neurons capable of elaborating neuropeptides and biogenic amines are an integral component of nervous systems. This apparatus is more extended than assumed during an earlier period of investigations. It is involved in short- and long-range communication by means of paracrine, transmitter-like, modulatory and neurohormonal types of messages. This finely adjusted activity of secretory neurons serves the control of a variety of important biological functions. Secretory pinealocytes are derivatives of pineal photoreceptors, primary sensory cells of neuronal character. In contrast to these neuron-like or paraneuronal elements, the secretory cells of the subcommissural organ are of ependymal origin. PMID- 3046225 TI - Circulating immune complexes in serum and in cerebrospinal fluid of patients with multiple sclerosis. Characterization and correlation with the clinical course. AB - We studied circulating immune complexes (IC) in the serum and cerebrospinal fluid (CSF) of patients with clinically defined multiple sclerosis (MS), in order to establish a correlation with the clinical course of the disease and to investigate the molecular composition of the IC isolated from patients in active phase of the disease. Serum IC levels were found to be significantly increased in patients from the progressive and active relapsing-remittent subgroups with both the CIC-conglutinin and C1q-binding methods. High levels of IC in CSF were detected only in the subgroup consisting of the relapsing-remittent patients in disease exacerbation when IC were determined by the C1q-binding test. No significant increase in serum or in CSF were found using the mRF-I test. The preliminary results of a qualitative investigation on serum IC in MS indicated that they are heterogeneous in nature, their size is mainly of the intermediate type, and they contain IgG, IgM, complement components and beta 2-microglobulins, the latter presenting an observation both new and interesting for studies on serum IC in MS patients. PMID- 3046226 TI - A controlled release form of madopar in parkinsonian patients with advanced disease and marked fluctuations in motor performance. AB - Flucuations in motor performance is a major problem in long-term levodopa treatment of Parkinsonian patients. A slow release preparation of levodopa with benserazide, Madopar HBS, has been developed in an attempt to decrease this problem. Eleven of 22 Parkinsonian patients with advanced disease and marked fluctuations experienced long-lasting benefit with reduction of their fluctuations in motor performance on treatment with Madopar HBS; 11 dropped out within the first 5 months of the trial. This was probably due to lack of experience with the effect of this new slow-release formulation. Nine patients (82%) required an additional dose of standard Madopar, especially in the morning. Significant improvements were found for akinetic phenomenon and dystonic cramps, and with the global evaluation of motor fluctuations. The occurrence of peak dose dyskinesia remained unchanged. No abnormalities in laboratory values were found. PMID- 3046227 TI - Congenital middle ear malformations. AB - After a profound review of normal embryology, a systematical and topographical classification of congenital middle ear malformations in general and of meatal atresia in particular is proposed; the classification of congenital aural atresia is essentially based on the recognition of two types according to the course of the facial nerve in its third segment: in type I, a normal topography of the facial nerve is found and consequently, the middle ear anomalies are rather minimal; in type II, an antero-superior displacement of the facial nerve in its third segment is recognized and the middle ear malformations are considerable worse. In this report, also attention is paid to syndromal nosology and its genetic aspects. In view of functional surgical; reconstruction, the technique of allograft canal surgery, similar to our combined approach tympanoplasty technique, is described in detail and compared with other classical techniques. Preoperative as well as postoperative problems and complications are extensively discussed. The excellent results of our technique are proposed even after 25 years of experience. An overview of cosmetic surgery techniques is given and also alternative techniques with implantable auditory prosthetic materials are summarized. PMID- 3046228 TI - Prevalence of Legionella in pharyngeal secretions of patients with pharyngitis. AB - A serological investigation has suggested that Legionella pneumophila may be associated with sore throat in adults. In a study of 177 adults and children with acute pharyngitis, Legionella species were not isolated from pharyngeal cultures, which utilized selective and nonselective buffered charcoal-yeast extract media. Group A beta-hemolytic streptococci were isolated from 14 percent of the 177 symptomatic patients. Throat cultures from 88 asymptomatic control subjects were negative for Legionella and beta-hemolytic streptococci. Further studies are needed to determine if Legionella species are associated with acute pharyngitis. PMID- 3046229 TI - Lacunar strokes: current concepts. AB - Lacunar strokes result from occlusion of penetrating arteries in the deeper, subcortical parts of the cerebrum and brain stem. Approximately 19 percent of all strokes are of the lacunar variety with lacunar strokes representing the most common cerebrovascular complication of chronic hypertension. Four major clinical syndromes are pure motor hemiparesis, pure sensory stroke, ataxic hemiparesis, and the dysarthria-clumsy hand syndrome. The advent of computed tomography (CT) has allowed the antemortem study of lacunar disease and has shed new light on the pathogenesis and clinical course of lacunar strokes. Recently, it has been demonstrated that lacunar strokes may be embolic or hemorrhagic in causation, are not invariably associated with hypertension, and may be larger and associated with hypertension, and may be larger and associated with neurological manifestations that do not conform to the classic patterns. In most instances, however, recognition of the characteristic clinical presentation and confirmation of the diagnosis with noninvasive studies spare many patients unnecessary risks associated with anticoagulation, arteriography, or vascular surgery. PMID- 3046230 TI - Evaluation and treatment of AIDS-associated illnesses: an approach for the primary physician. AB - The acquired immunodeficiency syndrome (AIDS) has become a problem of enormous clinical importance in the United States. This article describes the major clinical syndromes in adults with AIDS and offers approaches to their evaluation and treatment. PMID- 3046232 TI - Mortality and morbidity during a five-year follow-up of diabetics with myocardial infarction. AB - In 787 patients with acute myocardial infarction originally participating in the Goteborg Metoprolol Trial, mortality and morbidity during 5 years' follow-up were assessed and related to whether patients had diabetes mellitus. Diabetes occurred in 78 patients (10%). Patients with diabetes had a different risk factor pattern, including higher age, higher occurrence of angina pectoris and hypertension, whereas smoking habits did not differ. In the early phase (hospitalization), patients with diabetes had a higher mortality (12% versus 8%), required more treatment for heart failure and stayed longer in hospital. Other morbidity aspects, such as severity of pain, occurrence of severe supraventricular and ventricular arrhythmias, high-degree AV-block and infarct size did not differ. During 5 years' follow-up mortality rate in patients with diabetes mellitus was 55% as compared with 30% among patients with no diabetes (p less than 0.001). Reinfarction rate during 5 years was 42% in diabetics versus 25% in non-diabetics (p less than 0.001). In a multivariate analysis, taking into account the differences in risk factor pattern, diabetes turned out to be an independent determinant for long-term mortality and reinfarction (p less than 0.001). We conclude that patients with diabetes mellitus, developing acute myocardial infarction, is a group with particularly high risk of death and reinfarction. Interventions aiming at its reduction have priority. PMID- 3046231 TI - Office-based evaluation of renal function in elderly patients receiving nonsteroidal anti-inflammatory drugs. AB - Elderly patients with multiple diseases who are receiving diuretics are at risk for renal dysfunction from nonsteroidal anti-inflammatory drugs (NSAIDs). Fifty two elderly patients (mean age = 72 years, range = 63-87 years) with degenerative joint disease and multiple concomitant illnesses were randomly selected to receive ibuprofen suspension (400 mg) or aspirin (650 mg) 4 times a day. Serum creatinine (Cr), blood urea nitrogen (BUN), weight, and blood pressure were measured at baseline and at weekly intervals for 6 weeks. There were no significant changes from baseline in any tests reflective of renal function, no significant differences between ibuprofen and aspirin, and no influence of concomitant diuretic therapy. Ibuprofen and aspirin administered in the doses examined for 6 weeks appear to have little effect on renal function as measured by serum Cr and BUN in a sample of elderly patients for whom these drugs are commonly employed. Concomitant diuretic therapy does not appear to increase the risk. While these drugs are contraindicated in patients with severe hemodynamic insult, they should not be withheld from elderly patients who require this therapy for analgesic/anti-inflammatory effects because of concern for renal impairment. Further prospective research should be undertaken to clarify levels of patient risk and to define appropriate monitoring in such patients. PMID- 3046233 TI - Role of hypervolemia and renin in the blood pressure control of patients with pyelonephritis renal scarring. AB - Patients with pyelonephritic renal scarring are at risk of developing renal failure and hypertension. We studied glomerular filtration rate (GFR), renal plasma flow (RPF), filtration fraction (FF), systolic (SBP) and diastolic (DBP) blood pressure, fractional sodium, potassium and phosphate excretion, peripheral renin activity (PRA), plasma aldosterone (p-Aldo), urinary albumin excretion (U Alb) and urinary beta 2-microglobulin excretion (beta 2-M) in hydropenia and during transition to 3% volume expansion with isotonic saline infusion in 22 female patients with renal scarring due to pyelonephritis and 9 healthy controls. The patients had significantly lower GFR, higher SBP and higher PRA in hydropenia, but there was no significant difference in RPF, FF, DBP or p-Aldo. After volume expansion, SBP, DBP, PRA and p-Aldo were significantly higher in patients than in controls. Transition to 3% volume expansion was associated with a similar increase in SBP in both patients and controls, whereas DBP increased significantly more in the patients (p less than 0.01). Volume expansion resulted in a significant suppression of PRA and p-Aldo in both patients and controls. The patients with renal scarring had the same capacity to excrete sodium and water during transition to volume expansion as the healthy controls. The renin aldosterone system seems abnormally activated and is probably more important than hypervolemia in the development of hypertension in this group of patients. PMID- 3046234 TI - Insulin binds to specific receptors and stimulates macromolecular synthesis in C6 glioma cells. AB - The existence of insulin receptors and biological responses to insulin on macromolecular synthesis have been studied in C6 glioma cells. Binding of 125I insulin to C6 glioma cells was specific, time- and PH-dependent. Porcine insulin competed for 125I-insulin binding in a dose-dependent manner. Unlabeled polypeptides, including glucagon, bovine growth hormone, bovine prolactin did not compete for 125I-insulin binding. Scatchard analysis of the binding data gave a curvilinear plot which may indicate negative co-operativity or the existence of both high and low affinity (Ka = 7.55 x 10(10) - 4.25 x 10(9] sites. Incubation of cultures with insulin caused a time and dose-dependent stimulation of DNA, RNA and protein synthesis in C6 glioma cells (measured by 3H-thymidine, 3H-uridine or 3H-leucine incorporation into DNA, RNA, or protein respectively). The increase of macromolecular synthesis was admitted at more than 2 nM concentration of insulin. Maximal stimulation of DNA synthesis (142% of control) occurred 6 hours after incubation with 167 nM insulin. The same concentration of insulin caused a 45% increase in 1 hour on RNA synthesis, a 37% increase in 2 hour on protein synthesis. These results indicate that C6 glioma cells have specific insulin receptors capable of mediating effects of insulin on macromolecular synthesis. Insulin in the brain and even blood may be an important growth factor in the glioma cells of the patients with disrupted blood-brain-barrier. PMID- 3046235 TI - Trepanation as a therapeutic measure in ancient (pre-Inka) Peru. AB - Three cases of probably therapeutically intended trepanations of cranial fractures in ancient Peru are presented and discussed. PMID- 3046236 TI - Superimposition of an average three-dimensional pattern of brain structures on CT scans. AB - A method is described for the superimposition of an averaged telencephalic anatomical model and individual CT scans. The model is digitally available and derived from 3-D measurements of 30 post-mortem brains. It is averaged in size in relation to the midintercommissural point. The midintercommissural point in the individual brain is gained from reformed parasagittal projection images. The model is adjusted to the individual CT scan series by scaling and rotating according to the best fit and correspondence of the position of the central sulcus. The method needs no invasive neuroradiological techniques but is based on current computer algorithms. PMID- 3046237 TI - [Intestinal preparation for urological surgery: a review]. PMID- 3046238 TI - [Integrated urologic echography. I: Renoureteral pathology]. PMID- 3046239 TI - [Integrated urologic echography. II: Bladder pathology]. PMID- 3046240 TI - [Value of transrectal echography in the local staging of prostatic cancer]. PMID- 3046242 TI - [Removal and reimplantation of the isolated ureter in the rat]. PMID- 3046241 TI - [Nephrectomy of the heterotopic allograft]. PMID- 3046243 TI - [Monobactams]. PMID- 3046244 TI - [Percutaneous punctures and the kidney transplant]. PMID- 3046245 TI - Post-translational modifications of cellular proteins by polyamines and polyamine derivatives. PMID- 3046246 TI - Methylation, demethylation, and deamidation at glutamate residues in membrane chemoreceptor proteins. PMID- 3046247 TI - Perspectives on the biological function and enzymology of protein carboxyl methylation reactions in eucaryotic and procaryotic cells. PMID- 3046248 TI - Enzymatic methyl esterification of proteins and ageing: the eye lens as a model system for in vivo and in vitro studies. PMID- 3046249 TI - Post-translational methylations of ribosomal proteins. PMID- 3046250 TI - p34, a protein kinase involved in cell cycle regulation in eukaryotic cells. PMID- 3046251 TI - Evidence of protein kinase activity and characterization of substrate proteins in Escherichia coli. PMID- 3046252 TI - Post-translational modifications of the insulin receptor. PMID- 3046253 TI - The role of carbohydrate as a post-translational modification of the receptor for epidermal growth factor. PMID- 3046254 TI - Histone acetylation: a step in gene activation. AB - Cellular ageing appears to consist mainly in a loss of adaptability and a progressive decrease in the capacity of the cell to maintain homeostasis. Such age related phenomenon can be the result of stochastic or of programmed events, and may occur through changes in the base pairs or coding of the DNA, through increasing levels of error in transcription and finally through alterations at the translation step of proteins synthesis. The purpose of this chapter is to present histone acetylation as a key event in the control of chromatin structure and transcription. PMID- 3046255 TI - The molecular biology of influenza virus pathogenicity. PMID- 3046256 TI - Measurement of the affinity of antiviral antibodies. PMID- 3046257 TI - Vaccinia: virus, vector, vaccine. PMID- 3046258 TI - The pre-S region of hepadnavirus envelope proteins. PMID- 3046259 TI - Active therapeutic approaches to drug intoxication. PMID- 3046260 TI - Teratogenesis in vitro. PMID- 3046261 TI - [Sympathetic ophthalmia following vitrectomy and/or retinal detachment surgery]. PMID- 3046262 TI - [Histopathological and ultrastructural study of a case of leproma involving the entire cornea]. PMID- 3046263 TI - Hyponatremia and the brain. AB - Hyponatremia is a frequently encountered condition that requires careful management to avoid both complications of prolonged severe hyponatremia and life threatening brain damage resulting from too-rapid correction of the problem. Treatment before the condition becomes severe is the aim. Correction should be carried out as slowly as the clinical condition will allow; serum sodium should not be increased more than 12 mEq per L (12 mmol/per L) per day. Too-rapid correction may lead to demyelination syndromes. PMID- 3046264 TI - Use of psychiatric referral by family physicians. AB - The family physician occupies a critical place in the detection and management of mental health problems. Optimal case management requires an awareness of individual limitations and effective use of a psychiatric consultant. The consultant must understand the unique relationship between the patient and the family physician. Several historical biases may impede the physician's use of psychiatric consultation and the patient's acceptance of psychiatric treatment options. PMID- 3046265 TI - Acute high-altitude illness. AB - Acute high-altitude illness usually occurs at elevations of 8,000 ft or more, is more common than is generally appreciated and is often confused with other disorders. The acute forms of high-altitude illness include acute mountain sickness, high-altitude pulmonary edema, high-altitude cerebral edema and high altitude retinal hemorrhage. These illnesses often occur simultaneously and in severe forms can be fatal. Gradual ascent and acetazolamide help prevent acute mountain sickness and high-altitude pulmonary edema. The definitive treatment for all forms of high-altitude illness is descent to a lower altitude. PMID- 3046266 TI - Renal function in the elderly. AB - Renal mass, glomerular filtration rate and renal blood flow decline with age. Since the renal tubules also decrease in mass, the aging kidney is less able to compensate for changes in fluid balance. Because of changes in the renin aldosterone system, the elderly may have problems maintaining sodium and potassium balance. Physicians must be aware of the implications of these changes in renal function when they prescribe medications or order intravenous fluids for elderly patients. PMID- 3046267 TI - Polyorchidism. AB - Transverse division of the embryologic genital ridge explains the various forms of polyorchidism that have been reported. Most patients with supernumerary testes are asymptomatic and have painless groin or testicular masses. The majority have triorchidism, and the supernumerary testis is most frequently on the left side. The most common associated anomalies are inguinal hernia and maldescent of the testis. Testicular biopsy is essential for diagnosis. PMID- 3046268 TI - Pinworms. AB - The pinworm is one of the most common intestinal parasites in humans. The adult worms reside and mate in the area of the cecum and ascending colon. The females migrate to lay their ova in the perianal area. Perianal itching is the cardinal symptom of pinworm infestation. Collection of pinworms or ova from the perianal area permits diagnosis. Meticulous personal hygiene is essential to prevent spread of the parasites. PMID- 3046269 TI - Pseudofolliculitis barbae. AB - Pseudofolliculitis barbae is a chronic, distressing and potentially disfiguring dermatologic disorder that occurs predominantly in black men. The condition is caused by hairs curling back into the skin and is characterized by papules, pustules and, occasionally, keloidal scars over the beard region. Management includes cessation of shaving, the use of depilatories or topical antibiotics and modification of shaving techniques. PMID- 3046270 TI - Ototoxic drugs and the workplace. AB - Hearing conservation programs have not kept pace with changes in the workplace and in the work force. Heavy industry and noisy factories have yielded to quiet computers and high technology. Chemicals, self-administered or prescribed, may replace noise as a chief source of hearing loss. The aging working population and the elderly in general are vulnerable to ototoxicity. Conventional audiometric screening is no longer adequate. PMID- 3046271 TI - Hereditary multiple exostoses. PMID- 3046272 TI - Trousseau's syndrome. AB - Trousseau's syndrome refers to cancer-associated coagulopathy. The signs and symptoms of the syndrome are protean, and the underlying cancer is often occult. Treatment regimens vary. Heparin seems to be the initial drug of choice, perhaps continued indefinitely (intravenously or subcutaneously) on an outpatient basis. Warfarin does not seem to be beneficial. PMID- 3046274 TI - Management of overuse injuries. PMID- 3046273 TI - PCP: a dangerous drug. AB - PCP is again becoming a popular drug of abuse in the United States, particularly among the young. A variety of medical, psychiatric and pathologic effects make PCP both appealing and profoundly dangerous to naive and chronic drug users. Pharmacologic intervention is helpful in PCP-induced acute intoxication and psychosis. Effective long-term treatment is available in addiction programs. PMID- 3046275 TI - Propafenone: a new antiarrhythmic drug. PMID- 3046276 TI - Trauma: a systematic approach to management. AB - Initial care of the trauma victim begins at the scene of the accident, where the "load and go" principle is applied. A systematic approach to management of the trauma victim improves patient outcome. After insertion of an airway and stabilization of the spine, hemorrhage is treated and volume is replaced. At this point, invasive techniques, such as chest tube insertion, peritoneal lavage and pericardiocentesis, may be therapeutic as well as diagnostic. PMID- 3046279 TI - Noncardiac surgery in the cardiac patient. AB - As the population continues to age, more older patients present themselves for surgical procedures. Not uncommonly, these patients have considerable multiorgan dysfunction. Fundamental knowledge of predisposing risk is imperative for the optimal perioperative care of such patients. This review considers available data on factors that affect perioperative cardiac risk as they apply to specific clinical entities. PMID- 3046278 TI - Evaluation of a new inotropic agent, OPC-8212, in patients with dilated cardiomyopathy and heart failure. AB - OPC-8212 is a new positive inotrope with a unique mechanism of action. To assess its clinical utility we administered 60 mg/day for 30 days to 10 patients with overt congestive heart failure, who had substantial limitations in exercise performance despite treatment with conventional therapy. Patients were evaluated by simultaneous respiratory gas exchange and radionuclide ventriculography measurements during graded maximal exercise testing. Improvement was demonstrated in both peak oxygen uptake and radionuclide-determined cardiac index after 30 days of treatment with OPC-8212. These changes were not associated with alterations in heart rate at rest or during peak exercise. Furthermore OPC-8212 significantly decreased ventricular ectopy in our patients. However, two patients had possible hematologic toxicity with prolonged use. The results of the present phase II study suggest that a larger randomized double-blind placebo study to evaluate the safety and clinical efficacy of this drug is warranted. PMID- 3046280 TI - Paradoxical coronary embolism: case report and review of the literature. PMID- 3046277 TI - Comparison of the effects of chronic oral therapy with atenolol and sotalol on ventricular monophasic action potential duration and effective refractory period. AB - The effects of 4 weeks' therapy with atenolol, 50 mg twice daily, and sotalol, 160 mg twice daily, on ventricular monophasic action potential duration (MAPD90) and effective refractory period (VERP) are compared in a randomized cross-over study in 10 patients with stable angina pectoris undergoing elective cardiac catheterization. At matched ventricular pacing cycle lengths (range 500 to 1000 msec), MAPD90 was 29 to 42 msec (11.5% to 13.4%) longer and VERP was 21 to 32 msec (8.9% to 11.4%) longer on oral sotalol than on oral atenolol. Intravenous sotalol (100 mg) caused a significant lengthening of MAPD90 and VERP during oral atenolol therapy, while intravenous atenolol (10 mg) had no additional effect during oral sotalol therapy. The effects of sotalol on ventricular repolarization and refractory period persist over and above any adaptational response to beta receptor blockade that may occur during chronic therapy. PMID- 3046282 TI - Breast cancer: detection, prevention, and therapeutics. PMID- 3046281 TI - Pacemaker-induced superior vena cava syndrome: consideration of management. PMID- 3046283 TI - The German multicenter trial of anisoylated plasminogen streptokinase activator complex versus heparin for acute myocardial infarction. AB - A multicenter randomized trial of anisoylated plasminogen streptokinase activator complex (APSAC) versus heparin in patients with acute myocardial infarction of less than 4 hours' duration was undertaken in 19 hospitals. Of the 313 patients, 151 received heparin and 162 APSAC (30 U as intravenous injection). Within 28 days of hospital stay, 19 deaths (12.6%) occurred in the heparin group and 9 deaths (5.6%) in the APSAC group (p = 0.032). After 24 hours, patients in the APSAC group had a significantly lower incidence of cardiogenic shock (3.2 vs 9.5%, p = 0.031), asystole (3.8 vs 10.8%, p = 0.015) and need for resuscitation (5.1 vs 11.5%, p = 0.039). There was no difference in global and infarct-related ejection fraction between the 2 groups. Thus, APSAC favorably influences prognosis and clinical course in hospital. PMID- 3046284 TI - Comparison of coronary stenosis quantitation results from on-line digital and digitized cine film images. AB - To examine the effects of digital image acquisition mode and subtraction techniques on the results of coronary stenosis quantitation, 100 discrete lesions from 45 patients undergoing routine diagnostic angiography were analyzed in each of 3 image types: direct on-line digital, electrocardiogram-gated digital subtraction and digitized cine film images. For the geometric measurements (minimal lumen diameter and percent diameter stenosis) correlation coefficients for 2-way comparisons among the image types ranged from 0.90 to 0.96. Linear regression slopes ranged from 0.93 to 1.00, with intercepts from 0.03 to 0.07 mm for minimal diameter and -0.5 to 4.4% for percent diameter stenosis. For the videodensitometric percent area stenosis data, the correlation coefficients ranged from 0.80 to 0.89, with linear regression slopes from 0.84 to 0.89 and intercepts from 8.3 to 12.8%. Thus, the results of quantitative geometric measurements of coronary stenosis severity were not strongly affected by image acquisition mode (on-line versus cine film digitization) or by electrocardiogram gated digital subtraction, while densitometric data correlated less well when on line digital and digitized cine film acquisition methodology were compared. PMID- 3046285 TI - Fate of the pulmonic valve after proximal pulmonary artery-to-ascending aorta anastomosis for aortic outflow obstruction. AB - Transection of the main pulmonary artery and end-to-side anastomosis of the proximal pulmonary artery to the ascending aorta has been increasingly used in palliative surgery for cardiac malformations such as single ventricle with small outlet foramen (bulboventricular foramen) and hypoplastic left-heart syndrome. To evaluate pulmonary valve competence after this operation, we used color Doppler flow mapping to examine 45 survivors of pulmonary artery-to-ascending aorta anastomosis a median of 202 days postoperatively. Of 37 patients with hypoplastic left heart syndrome, mild regurgitation was detected in 9 (24%) and moderate regurgitation in 1 (3%). Of 8 with other lesions, mild regurgitation was observed in 2 and moderate regurgitation in 1. Seven of 11 patients imaged greater than or equal to 12 months postoperatively had regurgitation. In summary, one-fourth of survivors developed mild pulmonary regurgitation. Its presence should not be considered a contraindication to eventual application of Fontan's principle, although further follow-up appears warranted because the long-term fate of pulmonary valve function is not yet known. PMID- 3046286 TI - In appreciation of Alfred G. Mayer. PMID- 3046288 TI - Ultrasound angioscopy: real-time, two-dimensional, intraluminal ultrasound imaging of blood vessels. PMID- 3046287 TI - Saline contrast enhancement of trivial Doppler tricuspid regurgitation signals for estimating pulmonary artery pressure. PMID- 3046289 TI - Factors linking cholesterol to atherosclerotic plaques. PMID- 3046290 TI - Measurement of skeletal muscle blood flow in humans: plethysmographic, bolus and continuous infusion technique. AB - In this report a number of practical methods for the measurement of skeletal muscle blood flow in humans are presented and discussed. Special attention has been paid to the problems concerning representativeness for muscle blood flow (muscle vs other tissues and small muscle segments vs skeletal musculature of the total body) as well as the demand for steady state. PMID- 3046291 TI - Direct and indirect assessment of skeletal muscle blood flow in chronic congestive heart failure. AB - In patients with chronic congestive heart failure (CHF), skeletal muscle blood flow can be measured directly by the continuous thermodilution technique and by the xenon133 clearance method. The continuous thermodilution technique requires retrograde catheterization of the femoral vein and, thus, cannot be repeated conveniently in patients during evaluation of pharmacologic interventions. The xenon133 clearance, which requires only an intramuscular injection, allows repeated determination of skeletal muscle blood flow. In patients with severe CHF, a fixed capacity of the skeletal muscle vasculature to dilate appears to limit maximal exercise performance. Moreover, the changes in peak skeletal muscle blood flow noted during long-term administration of captopril, an angiotensin converting enzyme inhibitor, appears to correlate with the changes in aerobic capacity. In patients with CHF, resting supine deep femoral vein oxygen content can be used as an indirect measurement of resting skeletal muscle blood flow. The absence of a steady state complicates the determination of peak skeletal muscle blood flow reached during graded bicycle or treadmill exercise in patients with chronic CHF. Indirect assessments of skeletal muscle blood flow and metabolism during exercise performed at submaximal work loads are currently developed in patients with chronic CHF. PMID- 3046292 TI - Physiology of the renal baroreceptor mechanism of renin release and its role in congestive heart failure. AB - The function of the renal baroreceptor can be quantitatively described by a stimulus-response curve showing a flat section in the high pressure range, a steep slope in the low pressure range, and a well-defined threshold pressure slightly below resting systemic pressure. This stimulus-response curve shows a close functional relation to autoregulation of renal blood flow, glomerular filtration rate and sodium excretion. Threshold pressure and slope are subject to different physiologic control mechanisms: The slope is increased by a low sodium diet, whereas threshold pressure is elevated by an increased renal sympathetic nerve discharge or by circulating catecholamines. Sympathetic influences also reset renal autoregulation. Recent studies have provided evidence for an important role of the renal baroreceptor in the long-term control of arterial blood pressure. The sympathetic modulation of threshold pressure can induce sodium retention in early heart failure, and the sympathetic effects on autoregulation may help to explain clinical reports on a deterioration of renal function during converting-enzyme therapy. PMID- 3046293 TI - Skeletal muscle blood flow, metabolism and morphology in chronic congestive heart failure and effects of short- and long-term angiotensin-converting enzyme inhibition. AB - Blood flow to skeletal muscle during exercise is limited in patients with chronic congestive heart failure compared with normal persons. This may, in part, be attributed to the inability of the vessels to dilate adequately. In addition, an abnormal skeletal muscle metabolism with oxidative capacity can be demonstrated in patients with chronic congestive heart failure. The latter may partially explain why vasodilators and inotropes that improve central hemodynamics, global leg perfusion or skeletal muscle blood flow during exercise do not increase oxygen uptake by working muscle after acute drug administration. Furthermore, increases in blood flow to skeletal muscle may be redistributed to inactive or less oxygen-dependent muscle fibers. Although acute angiotensin-converting enzyme inhibition usually neither exerts substantial improvement in exercise hemodynamics nor interferes with leg perfusion during exercise, skeletal muscle blood flow and oxygen uptake gradually increases with long-term angiotensin converting enzyme therapy, leading to improved systemic oxygen consumption and exercise tolerance. Several factors, such as reduction in plasma and vascular angiotensin II and sympathetic nervous activity, increased prostaglandin levels and renal function may contribute to this beneficial long-term effect. PMID- 3046294 TI - Dependence of assessment of coronary artery reperfusion during acute myocardial infarction on angiographic criteria and interobserver variability. AB - The angiographic films of 240 patients with acute myocardial infarction were studied in a randomized trial of intravenous anisoylated plasminogen streptokinase activator complex (APSAC) versus intracoronary streptokinase therapies. The interobserver variability of grading coronary artery perfusion by the Thrombolysis in Myocardial Infarction Study Group (TIMI) criteria was measured as well as the effect of different definitions of reperfusion on the determination of reperfusion rate. There was good agreement in the reading of infarct artery flow grades between 2 blinded observers for each grade considered separately (k = 0.726 +/- 0.014) and for grades 0 or 1 (no perfusion) versus grades 2 or 3 (perfusion) (k = 0.905 +/- 0.011). Discordance between grades 0 or 1 versus 2 or 3 occurred in 74 (5%) of the 1,615 angiographic readings. Discrepancies of clinical significance which affected qualification for study entry, reperfusion or reocclusion status occurred in only 15 patients (6%). Grade 1 flow was found to have the most variable interpretation. Reperfusion rates for APSAC and streptokinase differed significantly when reperfusion was defined by 3 different criteria. The reperfusion rate ranged from 51 to 72% for APSAC and from 60 to 75% for streptokinase depending upon criteria selected. For comparison of the results of different thrombolytic studies, a standard semiquantitative system for grading infarct artery perfusion should be used, readings should be blinded and the criteria used for the definition of reperfusion should be clearly specified. PMID- 3046295 TI - Patterns of evolution of myocyte damage after human heart transplantation detected by indium-111 monoclonal antimyosin. AB - The indium-111 labeled Fab fragment of antimyosin monoclonal antibody was used to study cardiac rejection and the time course of myocyte damage after transplantation. Fifty-three studies were performed in 21 patients, 17 men and 4 women, aged 19 to 54 years (mean 37 +/- 8), from 7 to 40 months after transplantation. Repeat studies were available in 8, and 10 were studied after the first year of transplantation. A heart-to-lung ratio was used for quantitation of uptake (normal 1.46 +/- 0.04). Differences between absent (1.69 +/- 0.29) and moderate (1.90 +/- 0.36) rejection were significant (p less than 0.03). Antimyosin ratio at 1 to 3 months (1.89 +/- 0.35) differed from that at greater than 12 months (1.65 +/- 0.2) (p less than 0.01). Repeat studies revealed a decrease in antimyosin ratio in 5 patients with uneventful clinical course; 2 had persistent activity after transplantation and suffered heart failure from rejection. After 1 year of transplantation uptake was within normal limits in 7 of 10 patients, and high uptake was associated with vascular rejection in 1. Because they can define evolving patterns of myocardial lesion activity, antimyosin studies could be useful both in patient management and in concentrating resources for those patients who most require them. The heart-to lung ratio is suggested to monitor sequentially the degree of myocyte damage after transplantation. PMID- 3046296 TI - 1988 McCollum award lecture. The New Zealand selenium experience. PMID- 3046297 TI - Postprandial metabolic responses to the influence of food form. AB - To determine whether differences in the metabolic response to two common starches could be eliminated by altering the physical form of food, 12 normal and 6 noninsulin-dependent diabetic (NIDDM) subjects were studied after consumption of test loads of whole and blended rice and potato. In normal and NIDDM subjects the lower postprandial glycemia and insulinemia of whole rice was eliminated and became similar to that of whole potato, which was unaffected by blending. The glucagon responses were unchanged and similar in both groups under all study conditions. In both normal and NIDDM subjects the glucose and insulin response to a particular starch is not a stable feature dependent on the unique characteristics of the starch molecule but is affected by food processing and the form in which it is presented to the gastrointestinal tract. PMID- 3046298 TI - Compliance in a randomized clinical trial of dietary fat reduction in patients with breast dysplasia. AB - Dietary compliance was studied in 57 women participating for 1 y in a randomized clinical trial of dietary fat reduction. Nutrient analysis of food records, collected at 0, 6, and 12 mo, was compared with changes observed in lipid profiles and with chemical analysis of duplicate diets. Both food records and duplicate meals showed a significant decrease in fat intake (from 36 to 23% of total calories, p less than 0.0001) and a significant increase in carbohydrate (from 43 to 56% of total calories, p less than 0.0001) in the intervention group. The calculated nutrient intake from food records tended to overestimate the intake of protein, fat, and carbohydrate compared with the chemically analyzed method. The mean level of plasma cholesterol in the intervention group was significantly reduced (7.3%, p less than 0.01) in the first 6 mo after a reduction in dietary fat but the levels observed did not differ significantly between the groups at any time. PMID- 3046299 TI - Safety of oral intake of vitamin E. AB - A review of the literature concerning the safety of oral intake of vitamin E indicated that the toxicity of vitamin E is low. Vitamin E supplementation has resulted in inconsistent effects in serum lipid and lipoprotein levels. Animal studies showed that vitamin E is not mutagenic, carcinogenic, or teratogenic. In human studies with double-blind protocols and in large population studies, oral vitamin E supplementation resulted in few side effects even at doses as high as 3200 mg/d (3200 IU/d). PMID- 3046300 TI - Blood and urinary zinc changes after a glucose challenge in early and late pregnancies. AB - This study was undertaken to investigate the effect of pregnancy and glucose loading on zinc metabolism. In a completely random design with repeated measures, 18 non-pregnant women, 16 early-pregnant women (13-17 wk), and 16 late-pregnant women (28-34 wk) had blood collected at 0, 30, 60, 120, and 180 min after ingesting 100 g glucose to evaluate changes in variables of Zn nutriture. Fasting plasma Zn concentrations decreased significantly as pregnancy progressed. Late pregnant women had significantly higher erythrocyte Zn levels and greater 24-h urinary Zn and glucose excretions. Erythrocyte Zn responses to glucose loading were unaffected by gestational age. Plasma Zn after a glucose load in nonpregnant women exhibited a curvilinear response whereas pregnant women showed no change. This lack of response by pregnant women may be related to their lower plasma Zn concentrations. Plasma Zn in pregnant women may not be as readily available to assist in glucose utilization. PMID- 3046301 TI - Body fat distribution and hyperinsulinemia in childhood. PMID- 3046302 TI - Health aspects of vegetarian diets. AB - Recent studies of vegetarian diets and their effects on morbidity and mortality are reviewed. Vegetarian diets are heterogeneous as are their effects on nutritional status, health, and longevity. Mortality rates are similar or lower for vegetarians than for nonvegetarians. Risks of dietary deficiency disease are increased on vegan but not on all vegetarian diets. Evidence for decreased risks for certain chronic degenerative diseases varies. Both vegetarian dietary and lifestyle practices are involved. Data are strong that vegetarians are at lesser risk for obesity, atonic constipation, lung cancer, and alcoholism. Evidence is good that risks for hypertension, coronary artery disease, type II diabetes, and gallstones are lower. Data are only fair to poor that risks of breast cancer, diverticular disease of the colon, colonic cancer, calcium kidney stones, osteoporosis, dental erosion, and dental caries are lower among vegetarians. Reduced risks for chronic degenerative diseases can also be achieved by manipulations of omnivorous diets and lifestyles. PMID- 3046303 TI - Animal product consumption and mortality because of all causes combined, coronary heart disease, stroke, diabetes, and cancer in Seventh-day Adventists. AB - This report reviews, contrasts, and illustrates previously published findings from a cohort of 27,529 California Seventh-day Adventist adults who completed questionnaires in 1960 and were followed for mortality between 1960 and 1980. Within this population, meat consumption was positively associated with mortality because of all causes of death combined (in males), coronary heart disease (in males and females), and diabetes (in males). Egg consumption was positively associated with mortality because of all causes combined (in females), coronary heart disease (in females), and cancers of the colon (in males and females combined) and ovary. Milk consumption was positively associated with only prostate cancer mortality, and cheese consumption did not have a clear relationship with any cause of death. The consumption of meat, eggs, milk, and cheese did not have negative associations with any of the causes of death investigated. PMID- 3046304 TI - Vegetarian dietary practices and endurance performance. AB - Confounding influences of varying fat, protein, and carbohydrate (CHO) levels, training habits, and lifestyle patterns make the interpretation of specific influences of the diet on endurance performance unclear. In general, exhaustion during prolonged, hard endurance exercise is tied to low muscle glycogen stores. Athletes in heavy training are urged to consume 70% of calories as CHO to maximize body CHO stores. A deemphasis in animal products with an emphasis in high-CHO plant foods would facilitate athletes in conforming to nutritional recommendations. Some female athletes may increase their risk of iron deficiency and/or amenorrhea if a restrictive vegetarian diet is adopted. In general, the high-CHO nature of the vegetarian diet can help the endurance athlete in heavy training maximize body glycogen stores and thus the ability to perform. The balanced vegetarian diet provides the athlete with added reduction in coronary risk factors while meeting all known nutritional needs. PMID- 3046305 TI - Meat, starch, and nonstarch polysaccharides and large bowel cancer. AB - Starch and nonstarch polysaccharides enter the large gut where they are available for fermentation by the bacterial flora. The effect of fermentation on nitrogen metabolism, the supply of nutrients to the mucosa, and the carcinogen production is discussed in relation to epidemiological associations between dietary intake and large bowel cancer incidence. PMID- 3046306 TI - Role of bile acids and neutral sterols in carcinogenesis. AB - Epidemiological evidence regarding cancer causation suggests the existence of a strong link to diet-related lifestyles. Neutral sterols and bile acids constitute a group of metabolic endproducts known to have multiple interactions both from the standpoint of being influenced by diet as well as from the standpoint of their role in cellular and molecular processes relating to carcinogenesis. Epidemiological and experimental studies on the possible role of these steroid metabolites are reviewed. PMID- 3046307 TI - Role of dietary factors in cell replication and colon cancer. AB - Human studies and experimental data from animals suggest that high rates of colonic epithelial cell replication enhance the development of colon cancer. Vegetarians and individuals following a prudent diet have lower rates of colorectal cell proliferation than subjects at high risk for colon cancer. Animal studies show that colonic cell proliferation is stimulated by feeding in general and specifically by a number of dietary fibers, fats, bile acids, and short-chain fatty acids. Many of these growth factors also increase the induction of experimental tumorigenesis. On the other hand factors that reduce cell growth, including ascorbic acid and butylated hydroxyanisole, inhibit colon carcinogenesis. These results support the concept that dietary chemoprevention is feasible and could significantly reduce the rate of colon cancer development in high risk populations. PMID- 3046308 TI - Effect of diet on the plasma levels, metabolism, and excretion of estrogens. PMID- 3046309 TI - Vegetarian diet and blood pressure levels: incidental or causal association? AB - Evidence that nutrients other than the major cations may influence blood pressure levels stems from studies of acculturated vegetarians and from randomized controlled dietary trials. Earlier studies of vegetarians focused on religious groups and on vegans, making it difficult to know whether their lower blood pressures were due to diet per se or to other aspects of lifestyle. Seventh-day Adventist vegetarians showed significantly less hypertension and lower blood pressures compared with Mormon omnivores, effects which were independent of differences in obesity and not due to altered sodium intake. Subsequently, controlled dietary intervention studies in healthy normotensive omnivores provided more direct evidence for a blood pressure-lowering effect of a lactoovovegetarian diet with reversible changes of 5-6 mm Hg systolic and 2-3 mm Hg diastolic occurring over 6-wk periods. Similar dietary effects in mild hypertensive subjects provides impetus for identifying the responsible nutrients. PMID- 3046310 TI - Vegetarian children: appropriate and inappropriate diets. AB - Acceptable and appropriate vegetarian diets fulfill the Recommended Dietary Allowances and other authoritative dietary guidelines dealing with balance, variety, moderation, and developmental appropriateness of diets for children. Vegetarian regimes currently fed to infants and children are evaluated using these criteria. Vegan-like diets, fed early in infancy and childhood, pose special problems with respect to sufficiency of certain nutrients, energy, and bulk, especially if they are unplanned and unaccompanied by ongoing health supervision. Lactovegetarian, lactoovovegetarian, and semivegetarian patterns are more likely to be satisfactory. They conform closely with the pediatric recommendations for promoting health and reducing risks of chronic degenerative diseases, are sufficient without being excessive in nutrients, are low in bulk, and are developmentally appropriate. PMID- 3046311 TI - Food consumption, growth, and development of Dutch children fed on alternative diets. AB - A review of four studies examining food consumption growth and development of Dutch children fed on alternative diets is given. A literature study indicated that regarding child nutrition the three important movements in the Netherlands are the ecological movement, the anthroposophics, and the macrobiotics. A study on food consumption, height, and weight in preschool children fed these diets showed that the group of macrobiotic children were most at risk. Antropometric data collected in a cross-sectional study with 300 macrobiotic-fed children aged 0-8 y showed that the growth curves for boys and girls deviated from the Dutch standard curves after approximately 5 mo of age. There was no catch-up growth. In a selected sample of this latter group (43 children aged 4-6 y) mental development was measured by the Snijders-Oomen-Nonverbal intelligence test. The results of this test did not indicate an abnormal mental development for this age group of macrobiotic children. PMID- 3046312 TI - The Tromso Heart Study: diet, religion, and risk factors for coronary heart disease. AB - Low values of cholesterol and blood pressure in Seventh-day Adventists has been suggested to be caused by selection bias. Seventh-day Adventists in the Tromso Heart Study have a diet that is low in the intake of coffee, ground meat, and fish. They have total serum cholesterol values 11-20% (p less than 0.01) lower than non-Seventh-day Adventists. Blood pressure is also lower in Seventh-day Adventists. Baptists and religiously nonactive Seventh-day Adventists had a dietary pattern very similar to the general population. Their serum cholesterol and blood pressure values were also similar to the general population. This indicates that the low levels of serum cholesterol and blood pressure in religiously active Seventh-day Adventists is not caused by selective or religious factors. PMID- 3046314 TI - Vitamin B-12: plant sources, requirements, and assay. AB - Vitamin B-12 is of singular interest in any discussion of vegetarian diets because this vitamin is not found in plant foods as are other vitamins. Many of the papers in the literature give values of vitamin B-12 in food that are false because as much as 80% of the activity by this method is due to inactive analogues of vitamin B-12. PMID- 3046313 TI - Determinants of ischemic heart disease in Seventh-day Adventists: a review. AB - Most data from several countries shows Seventh-day Adventist men to have lower rates of ischemic heart disease (IHD) mortality. Similar data for women are somewhat conflicting. There is clear evidence that Adventists have lower serum total cholesterol and lower serum HDL cholesterol with the ratio of total cholesterol to HDL cholesterol being similar to that of non-Adventists. The risk relationships of this ratio may differ in different populations. There is a certain amount of evidence that vegetarians may have lower blood pressures but this is not clearly supported by data from Seventh-day Adventists. The lower risk for IHD in Adventist men, at least, is probably related to their dietary habits, nonsmoking status, possibly their better exercise habits, and greater social support. PMID- 3046315 TI - Mineral adequacy of vegetarian diets. AB - The bioavailability of a number of minerals may be altered by the special characteristics of vegetarian diets. Concern has centered on both inadequate and high dietary levels of specific minerals as well as reduced bioavailability because of a variety of dietary components. The possibility that plant-based diets may compromise mineral status is briefly reviewed for the following minerals: zinc, calcium, iron, manganese, selenium, and copper. PMID- 3046316 TI - Availability of essential amino acids and nitrogen in vegan diets. AB - Vegan children often fail to grow as well as their omnivorous cohorts despite protein intakes that exceed RDA. Explanations for inadequate growth include deficiencies of energy, calcium, zinc and vitamins B-12 and D. Due to decreased bioavailability, amino acids and nitrogen in vegan diets may be inadequate to support normal growth. Bioavailability of amino acids and nitrogen may be decreased by dietary fiber, food processing and storage, inadequate energy, and other unknown factors. Bioavailability should be considered when evaluating adequacy of intakes of protein, amino acids and nitrogen from vegan diets by infants and children. PMID- 3046317 TI - NCI dietary guidelines: rationale. AB - The National Cancer Institute (NCI) believes that the potential for dietary changes to reduce the risk of cancer is considerable and that the existing scientific data provide evidence that is sufficiently consistent to warrant prudent interim dietary guidelines that will promote good health and reduce the risk of some types of cancer. Six interim dietary guidelines and their scientific rationale are discussed herein. The evidence presented for the scientific rationale is based on the 1982 National Academy of Sciences Committee report Diet, Nutrition and Cancer and NCI's own scientific reviews that link long-term dietary patterns with cancer risk. These guidelines to the American public are consistent with other dietary recommendations from the US departments of Agriculture and Health and Human Services, the American Cancer Society, and the American Heart Association. PMID- 3046318 TI - Counseling the pregnant vegetarian. AB - Underlying any successful counseling effort must be an understanding of the acceptable and unacceptable foods of the particular person being counseled, with the recognition that nutrient needs may be met in many ways. This knowledge base necessary for successful counseling along with an understanding of other lifestyle practices that may have an impact on health will be difficult to develop unless rapport is established. Nonjudgmental, accepting attitudes on the part of the health professional are essential to the development of relationships with those individuals who may be suspicious of traditional medical and health care practices. Strategies for counseling with particular attention to the vegetarian are noted. PMID- 3046319 TI - Development of a quick reference guide to accommodate vegetarianism in diet therapy for multiple disease conditions. AB - With more Americans adopting vegetarian lifestyles, health-care professionals will encounter more patients whose vegetarian dietary practices must be reconciled with the therapeutic diets required by a variety of disease conditions. The development of a quick reference guide was undertaken to address several areas of concern unique to vegetarian patients and provide an expandable clinical resource. For the disease conditions examined, nutrition care goals were found to be met without compromising vegetarian practices. PMID- 3046320 TI - Food guides for the vegetarian. AB - Special consideration must be given to providing key risk nutrients in planning food guides for vegetarians especially if the diet chosen excludes all animal products, such as dairy foods and eggs. The practical use of food guides for education is also important. Several food guides for adult vegans, including pregnant vegan women, were analyzed by computer for nutritional adequacy using 7 d menus and comparisons were made with the RDA for energy, protein, iron, calcium, and riboflavin. Several patterns were identified that supply the RDA for protein, iron, calcium, and riboflavin and have educational merit. None of the patterns provide sufficient energy to meet the RDA. PMID- 3046322 TI - Quantitative immunohistochemistry: a new tool for surgical pathology? PMID- 3046321 TI - Chronic granulomatous disease of childhood. An unusual case of infection with Aspergillus nidulans var. echinulatus. AB - Aspergillus nidulans var. echinulatus was the sole agent cultured from the left lung, a paraspinal abscess, left ribs, and thoracic vertebral bodies from a patient with chronic granulomatous disease. Hyphal elements were present in histologic sections of lung, vertebral bodies, and infected ribs along with granuloma formation. The patient was treated with two debridement procedures and insertion of a Harrington rod followed by a long course of amphotericin B, flucytosine, and daily white blood cell transfusions. PMID- 3046324 TI - Foundations of immunohistochemistry. A practical review. AB - In this review of immunohistochemical methods, an emphasis is placed on the historic development of enzyme-linked procedures that are applicable to antigen localization in routinely processed tissues. As the technology of immunohistochemistry has improved, a bewildering montage of antibodies, bridges, labels, and chromogens has evolved. Each has its apparent advantages, yet each also has certain limitations. Admittedly, a comprehensive review of these different methods is beyond the scope of this forum, and several immunohistochemical techniques in current use will not be discussed in detail. Still, the principles of horseradish peroxidase-linked procedures, as described in this review, form the foundation of contemporary diagnostic immunohistology. They provide not only a powerful method of antigen detection, but also a model with which to compare recent and future innovations. PMID- 3046323 TI - Paul Ehrlich: the prototypic clinical pathologist. AB - The well-defined scientific roles of Paul Ehrlich are reviewed in context with modern concepts and practices in immunology. PMID- 3046325 TI - DNA probes in diagnostic pathology. AB - Molecular pathology has become firmly established as a distinctive discipline in medicine. It has introduced radical changes in concepts of disease causation and in classification of disease states affecting humans and other organisms. In addition, molecular pathology represents a "new" diagnostic technology with many potentials that have been heretofore untapped. This overview provides a discussion of the use of DNA probes in the study of human diseases. The role of detectable genetic abnormalities in pathogenesis will be considered, as well as their possible impact on nosology and disease classification. PMID- 3046326 TI - An interview with Arthur Hertig. Interview by Robert E. Scully. PMID- 3046327 TI - The Association's current logo and traditional seal: AAO emblems, symbols, and concepts. PMID- 3046328 TI - Shear bond strength of metal brackets compared with a new ceramic bracket. AB - One hundred twenty bovine teeth were bonded with two types of metal brackets and a new ceramic bracket for comparison. Two different adhesives were used, a so called no-mix and a paste/paste adhesive. The shear bond strength of the ceramic bracket was found to be superior for both adhesives. Bond failure with the ceramic bracket occurred predominantly in the enamel/adhesive interface; the failure site for the metal bracket was mainly in the bracket/adhesive interface. It is concluded that the bond strength between the ceramic bracket and the adhesive in shear mode is stronger than that between the adhesive and the enamel. PMID- 3046329 TI - Residual debris and bond strength--is there a relationship? AB - A study was undertaken to test the theory that there is a relationship between bond strength at the separate interfaces of the direct bonding system and the amount of residual debris remaining on the enamel surface following removal of a bonded appliance. In separate experiments, shear loads were applied at the complete direct bonding system and at the separate interfaces--namely, those at the adhesive/enamel and the adhesive/bracket base--using a chemically cured and a visible light-cured materials in combination with two designs of bracket bases. An assessment of residual debris following bond failure was made. For the complete direct bonding system, analysis showed that a foil mesh/chemically cured adhesive combination produces significantly more residual debris on the enamel than any other combination (P less than 0.05). No significant differences (P greater than 0.05) in shear bond strengths were detected for any bracket base/adhesive/enamel combination. Furthermore, no significant differences (P greater than 0.05) were detected between the shear bond strengths at the adhesive/enamel and adhesive/bracket base interfaces. These results suggest that the amount of residual debris following removal of the bonded bracket is not related to the shear bond strength at the separate interfaces but is governed by factors caused by bracket base design and properties of the adhesive used. PMID- 3046330 TI - Correlations between lower incisor crowding and lower incisor position and lateral craniofacial morphology. AB - Crowding of the lower incisors is a problem encountered frequently in orthodontic practice. Successful therapy may depend on the orthodontist's ability to evaluate factors contributing to the overall pattern. Two of these factors, vertical skeletal morphology and lower incisor position in the lateral cephalogram, were evaluated. Dental casts and cephalograms of 100 children with primary mandibular incisor crowding (that is, primary discrepancy between mesiodistal tooth width and available space of the dental alveolar process and apical base) were examined in this study. Results indicated the following: (1) all vertical skeletal and lower incisor position measurements closely duplicated published norms, (2) no correlation was found between lower incisor crowding and either skeletal morphology or lower incisor position, (3) a factor analysis did demonstrate, however, that other selected variables were interrelated, and (4) the cause of lower incisor crowding must be attributed to factors not examined in this study. PMID- 3046331 TI - Orthodontic bonding to porcelain--bond strength and refinishing. AB - Glazed porcelain surfaces are not amenable to resin penetration for orthodontic bonding. The aims of this study were to evaluate (1) the bond strengths of two orthodontic composite materials to treated porcelain surfaces, (2) the effect of thermocycling, and (3) the porcelain surfaces after refinishing. Sixty glazed porcelain disks were assigned to three surface treatment groups: Silane (S), roughening and silane (RS), and roughening (R). The acid-etched enamel surfaces of 20 extracted teeth served as a control group. Ten specimens in each group were bonded with either Concise or System 1. They were stored in water at 37 degrees C or thermocycled 8 degrees to 45 degrees C. Shear bond strength was tested with an Instron testing machine (0.05 cm/min). Analysis of variance showed that the four surface treatment groups and the two composite materials produced significantly different bond strengths (P less than 0.01 and P less than 0.05, respectively). The effect of thermocycling was not significant (P greater than 0.05). Acid etched enamel/Concise produced the highest mean bond strength (17.4 MN/m2). This was significantly greater (P less than 0.05) than the mean bond strengths of S/Concise (11.1), RS/System 1 (8.6), RS/Concise (8.1), and enamel/System 1 (7.8). The latter four mean bond strengths in turn were significantly greater (P less than 0.05) than S/System 1 (2.5), R/Concise (2.1), and R/System 1 (1.8). Of the four refinishing systems evaluated, all produced smooth surfaces but were unable to reproduce a glazed appearance. Roughening of porcelain and the silane treatment achieve bond strengths that should be clinically successful.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3046332 TI - Cyclosporine therapy for steroid-resistant nephrotic syndrome. A controlled study. AB - We have conducted a controlled trial on the efficacy of cyclosporine in eight patients with steroid-resistant nephrotic syndrome (four with idiopathic minimal lesion nephrotic syndrome and four with focal segmental glomerulosclerosis). Patients were randomly allocated to a cyclosporine (5 mg/kg/d) or a control group. After eight weeks of therapy and one month without cyclosporine therapy, patients in the control group were given cyclosporine for eight weeks and those in the cyclosporine group became controls. Before the initiation of treatment, there was no difference between the groups with regard to proteinuria and serum albumin levels. Proteinuria remained unchanged in the cyclosporine group, while there was a significant increase in proteinuria in the control group. There were no significant changes in serum albumin levels in either group during the trial. This study does not support the use of cyclosporine at the dose of 5 mg/kg/d in patients with steroid-resistant minimal lesion nephrotic syndrome or focal segmental glomerulosclerosis. PMID- 3046334 TI - Food allergy and the irritable bowel syndrome. PMID- 3046333 TI - Magnetic resonance imaging in the assessment of medullary compression in achondroplasia. AB - Children with achondroplasia may be at increased risk of developing apneic episodes and of dying unexpectedly. The risks seem to be related to neural axis compression by an abnormal cranial base and may be complicated by the development of hydrocephalus. We used magnetic resonance imaging to study five children with achondroplasia. All of them demonstrated a discrepancy between the size of the brain stem and the foramen magnum. Comprehensive prospective assessment of infants with achondroplasia, including the use of new imaging techniques, will provide important information concerning the natural history of the relationship of the neural axis to the bony posterior fossa and upper cervical spine in this condition. It may also help to identify those patients at risk before the development of life-threatening medullary compression. PMID- 3046335 TI - Generation of monoclonal antibodies to involved ileum of Crohn's disease: characterization of a panel of antibodies with goblet cell membrane and brush border-specific reactivity. AB - There are no known intestinal cell phenotypic markers characteristic of the transmural idiopathic inflammatory disease of the small bowel. To address this issue, we developed monoclonal antibodies specific for Crohn's disease tissue by generating a library of hybridomas specific for intestine following murine immunization with intestinal epithelial cells from a patient with Crohn's disease. The epithelial cells were initially harvested from a fresh operative specimen by gentle scraping and digestion with 1% collagenase on ice. Cells were then washed, evaluated for viability, and polytron homogenized. After immunization and subsequent fusion, hybridoma cell lines producing distinct antibody-binding patterns were identified by indirect immunoepifluorescence (IIEF). Wells representing distinct patterns were subcloned and carried to limiting dilution. Of the multiple hybridomas studied, the predominant target antibodies were goblet cell and glycoprotein-like molecules. Patterns of antibody binding identified by IIEF included: brush border, goblet cell, surface glycoprotein, enterocyte membranes, basilar crypt inclusions, circumferential goblet cell, and a heterogeneous goblet cell glycoprotein pattern. Molecular weight determination and cross-reactivity with various human tissues were studied by immunoblotting following sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. Several hybridomas were specific for the intestine but were not specific for Crohn's disease. PMID- 3046336 TI - Prevalence of gallstones in liver cirrhosis: a sonographic survey. AB - A sonographic prospective study of the prevalence of gallstones was performed in 140 patients with liver cirrhosis and in 140 controls. Gallstones were found more often in cirrhotic patients (29.2%) than in controls (13.6%) (p less than 0.01). Their prevalence increased with age. The ratio of women to men in cirrhotics was the same as in the general population, with a higher prevalence in women. The prevalence of gallstones increased in decompensated liver disease. There was a significantly higher prevalence of both hypersplenism and hemolysis in cirrhosis. No difference was found in gallstone prevalence in relation to cirrhosis etiology. This prospective study confirms, by means of sonography, the high prevalence of cholelithiasis in liver cirrhosis, and extends the previous data about the lithogenic risk factors in this disease. PMID- 3046337 TI - Measuring the separate effects of low parity and its antecedents on the incidence of ovarian cancer. PMID- 3046338 TI - Serum levels of selenium and retinol and the subsequent risk of cancer. AB - A nested case-control study was conducted to assess the relation between serum levels of selenium and retinol and the subsequent risk of cancer. During the years 1972-1984, in northwest Washington State, 156 cases of cancer were identified among members of two employee cohorts from whom specimens had been previously obtained and stored. Two hundred eighty-seven controls were selected from these cohorts and matched to cases on the basis of employer, age, sex, race, and date of blood draw. Selenium and retinol levels were measured by neutron activation and high pressure liquid chromatography, respectively. Information on known cancer risk factors was collected by telephone interviews of subjects and next of kin. Levels of selenium and retinol were unassociated with the incidence of cancer of all sites combined, both overall and within subgroups defined by age, sex, levels of the other micronutrient, time between blood draw and diagnosis, smoking status, and family history of cancer. These findings suggest that neither serum levels of selenium nor those of retinol have an appreciable effect on the risk of cancer. PMID- 3046339 TI - Prevalence of gallstone disease in a general population of Okinawa, Japan. AB - A total of 2,584 healthy residents in the Yaeyama District of Okinawa, Japan, were investigated in 1984 to determine the prevalence of gallstone disease and its associated factors. Diagnosis of gallstone disease was assessed by real-time ultrasonography. For participants over 20 years of age, obesity index and serum levels of total cholesterol and triglycerides were measured. Overall prevalence of gallstone disease was 3.2%. Prevalence increased with age from 0% under 19 years of age to 11.4% over 70 years of age and was higher in females (4.0%) than in males (2.5%). The results of the logistic regression analysis indicated that age and fatty liver were significant predictors of gallstone disease. The results of the automatic interaction detector analysis indicated that age and fatty liver were strong factors associated with gallstone disease and that prevalence was highest in females over age 50 with fatty liver. PMID- 3046340 TI - Significance and limits of cerebrospinal fluid beta-2-microglobulin measurement in course of acute lymphoblastic leukemia. AB - Cerebrospinal fluid beta-2-microglobulin (CSF-beta 2m) was measured longitudinally in 48 patients affected by acute lymphoblastic leukemia (ALL). Thirteen developed a central nervous system (CNS) involvement during the course of the disease; although moderately higher mean CSF-beta 2m levels were found in these subjects, no significant statistical differences were observed in comparison with patients without this complication and compared with the control group. No correlations were found between beta 2m and other biochemical parameters in CSF. Furthermore, CSF-beta 2m levels appeared to be influenced by previous combined chemoradiotherapeutic treatment for CNS prophylaxis, presence of meningeal non-neoplastic infiltrates, patients' ages, amount of CSF blasts, and their immunological phenotype. In particular, only clearly B-committed leukemic cells, when tested, showed a strong surface expression of beta 2m, as demonstrated by immunocytochemical detection of this protein on cell membrane. However, in specific cases, CSF beta 2m measurement and CSF/serum beta 2m ratio were helpful in diagnosing and monitoring isolated CNS disease. Such findings suggest that CSF-beta 2m assay may be a useful tool in the management of CNS involvement in the course of ALL in only selected patients, as several factors can modify the outcome. PMID- 3046341 TI - Immunohistochemical localization of renin in end-stage kidneys. AB - Hypertension in chronic renal failure is usually due to excessive accumulation of salt and water. In some cases, sodium and volume depletion by dialysis fail to reduce the high BP, and plasma renin activity tends to be higher. We performed a semiquantitative analysis of the immunohistochemical distribution of renin in the kidneys of ten patients with end-stage renal disease and hypertension using a specific antihuman renin antibody and a peroxidase-antiperoxidase technique on paraffin sections of nephrectomy and/or autopsy specimens. In five cases with severe, dialysis-resistant hypertension, the degree of immunoreactivity was most striking, exceeding that found in renovascular hypertension and present in arterioles at a distance from the glomeruli. Three cases of advanced diabetic glomerulosclerosis consistently showed minimal immunoreactivity. We conclude that renin often can be detected immunologically in the kidney of patients with chronic renal failure and hypertension, but its pathophysiological role will require further study. PMID- 3046342 TI - Loss of corticomedullary demarcation on magnetic resonance imaging: an index of biopsy-proven acute renal transplant dysfunction. AB - A prospective study of 19 cadaveric renal allograft recipients with suspected graft rejection was undertaken to compare the histological findings of the renal transplant biopsy with the results of magnetic resonance imaging (MRI). All 19 patients underwent a biopsy of the transplant allograft. Biopsy results included acute cellular rejection, acute vascular rejection, chronic vascular rejection (CVR), and acute tubular necrosis (ATN). Recipients of cadaveric renal allografts with normal function served as controls. The control showed distinct corticomedullary demarcation (CMD) on T1-weighted imaging. In contrast, CMD was absent or diminished in all the patients with suspected allograft rejection. Unfortunately, the loss of CMD did not correlate with a specific biopsy diagnosis. Patients with biopsy evidence of acute and chronic rejection or ATN demonstrated loss of CMD with similar image patterns. In conclusion, MRI is capable of detecting renal allograft dysfunction, but does not permit the determination of a specific cause. PMID- 3046343 TI - Tubular hypermetabolism as a factor in the progression of chronic renal failure. AB - Thus, in summary, we hypothesize that nephron loss by a time-limited insult (eg, an acute immunological insult, surgical removal of five sixths of normal nephrons) may cause events in the tubules of the remaining nephrons that contribute substantially, though not exclusively, to the inexorable progression to ESRD. Specifically, hypermetabolism in the remaining tubules as assessed by oxygen consumption and ATP synthesis rates has been found to occur. This hypermetabolism in residual nephrons appears to be due to the known increased sodium transport per nephron, but nonsodium transport events also appear to be involved. With respect to the latter phenomenon, we propose that an increase in growth factor response per tubule, as a mechanism of renal hypertrophy, activates the DAG----protein kinase C----Na/H antiporter system. We have evidence in support of this sequence of events by the demonstration of a two-fold increase in membrane (particulate) protein kinase C within 24 hours after removal of the contralateral kidney and an increase in intracellular pH as assessed by NMR spectroscopy. Activation of the Na/H antiporter not only leads to proton extrusion and cellular alkalinization, which may activate cellular alkalinization, which may activate cellular enzymes such as phospholipases, but also increases intracellular Na concentration, thereby further increasing Na/K ATPase, ATP use, and enhancing ATP synthesis. The increase in mitochondrial oxygen consumption, which accompanies the enhanced ATP synthesis, would be expected to be associated with increased oxygen radicle generation. If the tissue scavengers of oxygen radicles are not sufficient to scavenge the increase in oxygen radicles, then lipid peroxidation and tissue damage will occur.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3046344 TI - Testing for drug use, Part 1: Analytical methods. AB - Issues surrounding the screening and testing of individuals for drug use, including analytical and legal aspects of the procedures and social, political, and ethical problems and concerns, are reviewed. Historically, professional and societal debate regarding drug taking, drug-use problems, and the utility of drug testing programs occurs in cycles. Analytical methods commonly used to test for drug use include breath analysis for alcohol and urine drug assays. Blood alcohol concentrations are determined by laboratory assay methods or by portable devices used in the field. While poor laboratory procedures can invalidate test results for both breath and urine tests, urine screening test results can be further invalidated by improper handling of specimens or by tampering on the part of the subject. Also, test results are meaningful only if they are correlated with a clinical state. Legal issues have been raised concerning the validity of testing procedures used and the reliability of evidence obtained, especially in relation to pre-employment drug screening. From an ethical standpoint, drug testing tends to focus efforts to combat drug abuse on the drugs themselves instead of on the social context of the problem. With a recycled interest in drug-use testing and screening, primarily attributable to technological advances, little attention is being given to other approaches to controlling drug use. Additional research is needed to better describe the nature and extent of our drug-use problems and their impact on society. PMID- 3046345 TI - Continuous arteriovenous hemofiltration: an alternative to hemodialysis. AB - An introduction to the procedure of continuous arteriovenous hemofiltration (CAVH) for management of acute renal failure, as well as a review of hemodialysis, is presented. Initially developed for the management of hemodynamically unstable patients with acute renal failure, CAVH now is also used for management of fluid overload and acid-base disturbances resulting from conditions such as acute pulmonary edema, congestive heart failure, septic shock, and oliguric states in which pharmacologic or parenteral nutrition therapy necessitates administration of large volumes of fluids. CAVH, in contrast to hemodialysis, does not typically involve use of blood pumps but uses the patient's own mean arterial pressure to generate a driving force across the hemofilter membrane. CAVH, like hemodialysis, can remove excess fluid and uremic toxins; however, fluid removal by CAVH is characterized by the slow, continuous process of ultrafiltration and thus avoids the risk of hypotension, muscle cramps, or disequilibrium syndrome. Furthermore, CAVH does not require fluid restriction, allowing for increased administration of parenteral nutrition and intravenous medications; neither does it require expensive equipment or highly trained personnel. Although CAVH membrane materials may differ, they all permit the removal of plasma water and non-protein-bound solutes with molecular weights less than 10,000. To prevent blood from clotting in the hemofilter, most patients will require administration of heparin, which in some patients may increase the possibility of hemorrhaging. CAVH also can remove pharmacologic agents from the blood; however, only the non-protein-bound fraction of the drug has the potential to be cleared from the bloodstream by CAVH.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3046346 TI - Testing for drug use, Part 2: Legal, social, and ethical concerns. PMID- 3046347 TI - Sensitivity and specificity of three methods of detecting adverse drug reactions. AB - The sensitivity and specificity of three methods of detecting adverse drug reactions (ADRs) were determined. Minimal use of a voluntary ADR reporting program prompted this investigation of three ADR detection methods, as follows: screening of laboratory reports, pharmacist screening of medication orders, and voluntary reporting. A total of 98 patients who were receiving oral or i.v. digoxin therapy, oral or i.v. theophylline therapy, or i.v. gentamicin therapy were randomly selected and monitored for possible ADRs. A physician reviewed the charts of patients with suspected ADRs using the Naranjo algorithm to assess causality. The chart review served as the reference method to which the other three methods were compared. Thirteen "true" (i.e., confirmed by the Naranjo algorithm) ADRs were identified in 11 different patient charts, resulting in a 13.3% ADR incidence rate for the 98 sampled patients. For the three ADR detection methods, the decreasing order for level of sensitivity was screening of laboratory reports, pharmacist screening of medication orders, and voluntary reports; however, only the difference between laboratory reports and voluntary reports was significant. For level of specificity, the decreasing order for the three methods was voluntary reports, pharmacist screening of medication orders, and pharmacist screening of laboratory reports; the differences among all three methods were significant. Screening of laboratory reports and pharmacist screening of medication orders are two detection methods that appear to exhibit an appropriate combination of sensitivity and specificity for identifying ADRs; trials with larger sample sizes are needed to confirm the results of this study. PMID- 3046348 TI - Compliance with a restricted antimicrobial agent policy in a university hospital. AB - Compliance with a policy for use of antimicrobial agents that requires both oral approval from the infectious diseases service and completion of a restricted antimicrobial agent use form was evaluated in a 950-bed teaching hospital. The charts of patients for whom a restricted antimicrobial agent was ordered during four one-week periods between January 1987 and April 1987 were audited to determine whether completed use forms had accompanied orders for restricted antimicrobial agents. The validity of the information on completed forms was determined by comparing the information on the form with notes in patients' charts and through discussions with infectious diseases physicians. Two infectious diseases physician reviewers evaluated the appropriateness of prescribing of piperacillin and ceftazidime by analyzing data collected by pharmacists. Forms were submitted with 132 of 154 orders written for restricted agents; incomplete forms were received and accepted by pharmacists for 39 courses of therapy. The infectious disease service had not been contacted to approve use of a restricted agent in 25 cases. Eight of the 48 courses of piperacillin or ceftazidime therapy were deemed inappropriate despite initial infectious diseases approval. Compliance with a formal antimicrobial agent restriction policy at this institution has been good, but periodic re-education and follow-up monitoring appear to be necessary to ensure optimal use of restricted agents. PMID- 3046349 TI - Impact of a pharmacokinetics consultation service on clinical outcomes in an ambulatory-care epilepsy clinic. PMID- 3046350 TI - Intravenous acyclovir therapy of first episodes of genital herpes: a multicenter double-blind, placebo-controlled trial. AB - PURPOSE: A collaborative multicenter double-blind, placebo-controlled trial of intravenous acyclovir treatment of first-episode genital herpes was performed in order to substantiate previous findings on the efficacy and safety of this drug, to evaluate the influence of parenteral therapy on recurrence frequency, and to obtain further data on the natural history of genital herpes. PATIENTS AND METHODS: Eighty-two patients with first episodes of genital herpes simplex virus (HSV) infection were randomly assigned in a double-blind fashion to treatment with intravenous acyclovir (5 mg/kg every eight hours) or placebo for five days. Before therapy, all lesions in the genital/perineal area and in extragenital sites were cultured. New lesions appearing in both areas after the onset of therapy were cultured separately. Lesions in all groups were cultured until completely healed. Sera were collected from all patients on entry to the study and on Day 21 to determine presence or absence of antibodies to HSV-1 and HSV-2. Time to healing, time to crusting, time to cessation of viral shedding, and appearance of new lesions during therapy were compared for each treatment group. RESULTS: Patients receiving acyclovir experienced a significant reduction in the median duration of pain (4.3 versus 4.8 days, p = 0.019), viral shedding (1.9 versus 8.4 days, p less than 0.001), and time to healing (8.4 versus 11.5 days, p = 0.02) compared with placebo recipients. These differences were largely attributable to the effect of therapy in the subset of patients with primary disease in whom acyclovir reduced the median duration of pain from 10.6 days to 4.2 days, the median duration of viral shedding from 17.1 days to 1.9 days, and the median time to healing from 14.2 days to 8.3 days. The rate of subsequent recurrence of genital herpes was not altered by acyclovir treatment: 24 of 32 acyclovir recipients (75 percent) experienced one or more recurrences during a mean follow-up of 14 months compared with 19 of 27 placebo recipients (70 percent). Among patients experiencing recurrences, the mean number of recurrences per month among acyclovir recipients was 0.25 compared with 0.19 for patients given placebo. CONCLUSION: This multicenter trial confirms the efficacy of intravenous acyclovir in the management of first-episode genital herpes, especially in patients with primary infection. However, therapy did not alter the frequency of recurrences. PMID- 3046351 TI - An attachable silver-impregnated cuff for prevention of infection with central venous catheters: a prospective randomized multicenter trial. AB - PURPOSE: Percutaneously inserted central venous catheters are widely used. Catheter-related bacteremia or fungemia is the most frequent serious complication of these catheters. In an attempt to reduce the frequency of such infections, a subcutaneous cuff constructed of a biodegradable collagen matrix impregnated with bactericidal silver was developed. Our goal was to assess, in a multicenter clinical trial, the effectiveness of this cuff in preventing catheter-related infection. MATERIALS AND METHODS: Central venous catheters needed for fluid or drug therapy, hemodynamic monitoring, or hyperalimentation in patients in three centers were randomly assigned to be inserted with or without the cuff. Patients and catheters in the two groups were comparable in terms of risk factors predisposing to infection, including colonization of skin about the insertion site. RESULTS: The results with 234 catheters inserted into a new site showed that catheters inserted with the cuff were threefold less likely to be colonized on removal (more than 15 colony-forming units) than were control catheters (28.9 percent versus 9.1 percent, p = 0.002) and were nearly fourfold less likely to produce bacteremia (3.7 percent versus 1.0 percent). Adverse effects from the cuff were not seen. The cuff did not confer protection, however against infection with catheters inserted over a guidewire into old sites. Most of the catheter related infections identified in this study, including four of the six bacteremias, appear to have been caused by microorganisms colonizing skin about the insertion site, affirming the pathogenetic basis for benefit seen with the cuff in this clinical trial; two may have derived from contamination of the catheter hub. CONCLUSION: This novel, silver-impregnated, attachable cuff can substantially reduce the incidence of catheter-related infection with most percutaneously inserted central venous catheters, can extend the time catheters can be left in place safely, and can prove cost-beneficial. PMID- 3046352 TI - Treatment of proteinuric idiopathic glomerulonephritides in adults: a retrospective survey. AB - PURPOSE: A review of the worldwide medical literature was undertaken to determine whether treatment with currently available drugs has been beneficial in patients with glomerulonephritides. MATERIALS AND METHODS: Thirty-five articles, involving 1,653 adult patients with chronic primary glomerulonephritis with proteinuria and/or nephrotic syndrome, were analyzed. The criteria for inclusion of data were strict, and only complete and well-studied cases were included in the analysis. The papers were grouped into prospective controlled trials, retrospective comparisons of treated and untreated patients, and retrospective studies without an untreated group. RESULTS AND CONCLUSIONS: Analysis of the data shows that patients with minimal change disease clearly benefited from the administration of corticosteroids and/or cytotoxic drugs, since 89 percent and 79 percent of patients had a complete remission in controlled trials and in retrospective studies, respectively. A low incidence of complete and partial remissions was found in the group of patients with focal segmental glomerulosclerosis, proteinuria disappeared in 19 percent, and only 36 percent of these patients had stable renal function after a mean follow-up period of 6.3 years. The incidence of uremia and the number of patients requiring dialysis were very high. In patients with membraneous glomerulonephritis, conflicting results were obtained in controlled trials, retrospective comparisons, and retrospective studies, since a high percentage of complete and partial remissions was present in treated patients from both controlled and retrospective comparisons, whereas the percentage of complete and partial remission in untreated patients increased as the length of the follow-up period increased. Long-term studies carried out on untreated patients confirmed the high percentage of remissions. These data suggest that it is necessary to await future long-term results from controlled trials in order to determine whether or not corticosteroids alone or associated with cytotoxic drugs have a beneficial effect. Finally, the data from patients with mesangiocapillary glomerulonephritis show that therapy appears to have no beneficial effect on renal function. Nephrologists need new drugs that interfere more decisively with the immune process in the pathogenesis of primary glomerulonephritis. PMID- 3046354 TI - Malignant external otitis: insights into pathogenesis, clinical manifestations, diagnosis, and therapy. AB - Malignant external otitis is an infection of the external ear canal, mastoid, and base of the skull caused by Pseudomonas aeruginosa. The condition occurs primarily in elderly patients with diabetes mellitus. Current theories on pathogenesis and anatomic correlations are reviewed. Severe, unrelenting otalgia and persistent otorrhea are the symptomatic hallmarks of the disease, whereas an elevated erythrocyte sedimentation rate is the only distinctive laboratory abnormality. Iatrogenic causes such as administration of broad-spectrum antibiotics and aural irrigation may play a predisposing role in high-risk populations. The disease can result in cranial polyneuropathies (with facial nerve [VII] paralysis being the most common) and death. The mainstay of treatment is administration of antipseudomonal antibiotics for four to eight weeks. Recurrence is common, and mortality remains at about 20 percent despite antibiotic therapy. Given the increasing longevity of diabetic patients, the frequency of this disease is increasing. Internists, family practitioners, and ambulatory care physicians must now be cognizant of the presenting symptoms, while infectious disease specialists and otolaryngologists need to be appraised of strides in diagnosis and therapy. The role of surgery should be minimized. Use of new diagnostic radiologic modalities and new antipseudomonal antibiotics discussed in this review should lead to improved outcome. PMID- 3046355 TI - Medical informatics: an introduction to computer technology in medicine. AB - Access and effective management of medical information have become increasingly important in the practice of medicine today. Computer technology is developing to achieve this goal. This had led to the emergence of a new specialty, medical informatics, the basic science of the use of computers in medicine. Areas of patient care to which medical informatics has been applied include history taking, medical records, medical data base information retrieval, test performance, test result retrieval, decision support, patient monitoring, medical education, quality assurance and utilization review, medical research, and medical office and financial management. It is important that these applications become integrated with existing medical information systems and that physicians take a leading role in developing and maintaining these systems. PMID- 3046353 TI - Captopril in patients with type II diabetes and renal insufficiency: systemic and renal hemodynamic alterations. AB - PURPOSE: To our knowledge, clinical studies on the long-term use of angiotensin converting enzyme inhibitors in patients with type II diabetes mellitus and nephropathy with incipient renal failure are nonexistent. We therefore assessed the effects of long-term treatment with captopril on systemic and renal hemodynamics and urinary protein excretion in patients with type II diabetes mellitus and the clinical syndrome of diabetic nephropathy. PATIENTS AND METHODS: Twelve patients, 10 men and two women, with an average age of 52 years (range, 40 to 66), participated in the study. After the basal hemodynamic evaluation, the patients received captopril in two daily doses. The dosage was adjusted at weekly intervals in order to obtain normalization of blood pressure without exceeding the maximum allowable dosage. Four patients also received furosemide (20 to 40 mg/day). RESULTS: After six months of treatment, the intra-arterial blood pressure fell (from 162 +/- 17/103 +/- 5 to 139 +/- 26/89 +/- 10 mm Hg) due to the reduction in total peripheral vascular resistance index (from 3,720 +/- 658 to 3,190 +/- 762 dynes/second/cm-5/m2), with no change in cardiac index (2.78 +/- 0.36 to 2.79 +/- 0.47 liters/minute/m2). No significant change was seen in renal vascular resistance (from 30,175 +/- 5,371 to 30,173 +/- 5,372 dynes/second/cm 5/1.73 m2) and in filtration fraction (from 26 +/- 8 to 27 +/- 10 percent). A slight, not significant, decrease in renal plasma flow (from 243 +/- 97 to 217 +/ 108 ml/minute/1.73 m2), in glomerular filtration rate (from 57 +/- 17 to 51 +/- 19 ml/minute/1.73 m2), and in proteinuria (from 4.50 +/- 3.10 to 3.40 +/- 2.31 g/day) was also observed. CONCLUSION: Our findings suggest that captopril is an effective antihypertensive agent in patients with diabetic nephropathy, but the renal beneficial effects seem to be limited when this syndrome is complicated by renal insufficiency. PMID- 3046356 TI - Pellagra: a revisionist approach. PMID- 3046357 TI - Toxoplasma orchitis: report of a case and a review of the literature. PMID- 3046358 TI - Synergistic nephrotoxicity due to ciprofloxacin and cyclosporine. PMID- 3046359 TI - Profound, yet reversible, heart failure secondary to 5-fluorouracil. PMID- 3046360 TI - Is behavioral modification a sufficient criterion of the effectiveness of decision analysis? PMID- 3046361 TI - Cardiac rehab: still running? or standing still? PMID- 3046362 TI - Characteristics of convergence insufficiency. AB - Convergence insufficiency (CI) is a well-known syndrome of binocular visual dysfunction. In a review of 58 papers, considerable variation was noted in the criteria used to define the condition. Symptoms and decreased positive fusional vergences (both at the nearpoint) were the only criteria named in more than one half of the studies. An extended nearpoint of convergence (NPC) and increased exophoria at near were criteria in about one-third of the papers. Examination of data in the reviewed papers shows that although considerable variability was noted, the distance exodeviation, distance negative vergences, visual acuity, refraction, and stereopsis were about the same as population norms. Positive vergences, negative vergence at near, and NPC were somewhat less than population norms. Vergences relative to Sheard's criterion, the near exophoria, accommodative amplitude, and the AC/A ratio were consistently below those derived from population norms. This considerable variation may largely be a result of different criteria used for diagnosis, subpopulations within the data, and other confounding factors. PMID- 3046363 TI - Visual therapy results for convergence insufficiency: a literature review. AB - This paper is a review of the literature relative to treatment results for convergence insufficiency utilizing vision therapy training procedures. Vision therapy is shown to improve the nearpoint of convergence and fusional convergence and to ameliorate associated symptoms. The overall cure rate is 72%. Furthermore, the training results appear to persist for at least 2 years if the patients are initially cured and are independent of age until the late presbyopic years. Also, recent studies indicate the type of training procedures which yield the most effective training results. PMID- 3046364 TI - Review of computerized orthoptics with specific regard to convergence insufficiency. AB - Traditional vision training or orthoptics has used line or contour targets to eliminate suppression and improve vergence performance. Manipulation of these stimuli is slow and arduous. Line stimuli require an experienced doctor/technician to interpret responses. Recently, automated vision training using microprocessor anaglyph stimuli, i.e., random dot stereograms (RDS), has been used in an operant conditioning paradigm. This technique has improved motivation of the patient, improved reliability, and provided standardization of therapy. In addition, the utilization of RDS associated with operant conditioning has been shown to improve vergence performance and to reduce asthenopia in the convergence insufficiency patient. PMID- 3046365 TI - Influence of accommodative and vergence adaptation on binocular motor disorders. AB - Tait described four categories of binocular disorders including convergence excess, convergence insufficiency, divergence excess, and divergence insufficiency. These disorders are defined by the distance where the largest heterophoria occurs (distance or near), and the amplitude of the accommodative vergence ratio (AC/A). Insufficiency corresponds to a low AC/A ratio, whereas excess corresponds to a high AC/A. The magnitude of the AC/A ratio, which may be influenced by the adaptability of the accommodation and vergence systems, has been shown to be reciprocally related to adaptability of accommodation. Likewise, the degree of vergence accommodation has been shown to be related reciprocally to adaptability of vergence to prism. An imbalance of adaptability of accommodation and vergence systems produces abnormal cross-coupling between the two motor systems. When accommodation is more adaptable than vergence, the AC/A ratio is low and the CA/C ratio is high. Conversely, when vergence is more adaptable than accommodation, the AC/A ratio is high and the CA/C ratio is low. A method is reported for temporarily restoring moderate amplitudes of abnormal AC/A and CA/C ratios by reducing excessive adaptation with fatigue. Finally, new clinical procedures for measuring adaptation of accommodation and the CA/C ratio are presented. Taken together with current measures of vergence adaptation and AC/A ratio these procedures will permit a more complete evaluation of mutual interactions between accommodation and vergence in patients diagnosed as having excessive and insufficient vergence. PMID- 3046366 TI - Evaluation of contact lenses by microbial enumeration and protein determination. AB - Contact lenses worn for varying periods of time (from 1 to 48 months) were subjected to microbiological examination by plate counts and protein determination. Seventy percent of the lenses displayed bacterial colony counts below 120 colony-forming units (CFU)/lens, 28 percent were in the range of 140 to 9060 CFU/lens, and one lens was contaminated with greater than 6 x 10(4) CFU/lens. Fungal contaminants were detected in three lens specimens in the range of 220 to 760 CFU/lens. Protein accumulation showed wide variation of up to 1.2 mg of protein per lens. Statistical analysis indicated highly significant associations (p less than 0.001) between the bacterial colony counts obtained with three different media. Some significant associations were found between the protein concentration and bacterial counts. The data did not indicate statistically significant relations between the above variables and either the water content or the length of wear of the contact lenses. PMID- 3046367 TI - Immunocytochemical study of dystrophin in muscle cultures from patients with Duchenne muscular dystrophy and unaffected control patients. AB - Using immunocytochemical methods, the localization of dystrophin, the gene product affected in Duchenne muscular dystrophy (DMD) in aneural, differentiating human muscle cultures, was studied. Dystrophin was not demonstrable in undifferentiated myoblasts from control patients and from two patients with DMD. After myoblast fusion, the protein was found in circumscribed sarcoplasmic patches, in the perinuclear area, and along the surface of all normal multinucleate myotubes, with more mature myotubes showing predominantly sarcolemmal distribution. There was no staining in myotubes from one DMD patient and only faint diffuse fluorescence in myotubes from the second affected boy, however. These data provide further evidence that dystrophin is a sarcolemma associated protein, that it is developmentally regulated, and that it is absent or greatly reduced in quantity in skeletal muscle cultures from patients with DMD. PMID- 3046370 TI - Human temporal bone from the lower Paleolithic site of Castel di Guido, near Rome, Italy. AB - The Castel di Guido site, with an estimated age of approximately 300,000 years, has yielded abundant animal remains, Acheulian stone and bone bifaces, and small tools. On the surface of the original deposit, turned over by agricultural activities, fragments of human remains were discovered between 1980 and 1986, including a temporal bone described here. PMID- 3046368 TI - Perturbation of cultured human endothelial cells by atherogenic levels of low density lipoprotein. AB - Cultured human umbilical vein endothelial cells (EC) exposed to atherogenic levels of low density lipoprotein (LDL) for protracted periods demonstrated no measurable evidence of overt cytotoxicity, but were perturbed as indicated by an increase in prostacyclin (PGI2) production. Confluent EC were incubated with high LDL concentrations (240 or 330 mg/dl cholesterol) for 1 to 12 days. LDL was added to culture media containing 25% human lipoprotein-deficient serum to determine the effects of LDL independent of other lipoproteins. LDL did not injure EC as assessed by cell count, vital dye exclusion, 51chromium release, and lactate dehydrogenase release. Although high concentrations of LDL did not cause EC cytotoxicity, such LDL concentrations did results in increased PGI2 generation. PGI2 accumulation in postincubation media was increased two-to-fivefold in otherwise unstimulated cells as measured by radioimmunoassay of the stable PGI2 breakdown product, 6-keto-PGF1-alpha. This elevation persisted for the entire 12 day exposure to high LDL concentrations. These results indicate that prolonged exposure to atherogenic concentrations of LDL does not effect EC viability, but does cause an endothelial perturbation as demonstrated by an increased PGI2 production. PMID- 3046369 TI - Antibody-mediated rejection of human orthotopic liver allografts. A study of liver transplantation across ABO blood group barriers. AB - A clinicopathologic analysis of liver transplantation across major ABO blood group barriers was carried out 1) to determine if antibody-mediated (humoral) rejection was a cause of graft failure and if humoral rejection can be identified, 2) to propose criteria for establishing the diagnosis, and 3) to describe the clinical and pathological features of humoral rejection. A total of 51 (24 primary) ABO-incompatible (ABO-I) liver grafts were transplanted into 49 recipients. There was a 46% graft failure rate during the first 30 days for primary ABO-I grafts compared with an 11% graft failure rate for primary ABO compatible (ABO-C), crossmatch negative, age, sex and priority-matched control patients (P less than 0.02). A similarly high early graft failure rate (60%) was seen for nonprimary ABO-I grafts during the first 30 days. Clinically, the patients experienced a relentless rise in serum transaminases, hepatic failure, and coagulopathy during the first weeks after transplant. Pathologic examination of ABO-I grafts that failed early demonstrated widespread areas of geographic hemorrhagic necrosis with diffuse intraorgan coagulation. Prominent arterial deposition of antibody and complement components was demonstrated by immunoflourescent staining. Elution studies confirmed the presence of tissue bound, donor-specific isoagglutinins within the grafts. No such deposition was seen in control cases. These studies confirm that antibody mediated rejection of the liver occurs and allows for the development of criteria for establishing the diagnosis. PMID- 3046371 TI - Age and sex biases in the preservation of human skeletal remains. AB - Inaccuracies introduced through biases in preservation are a major source of error in paleodemographic reconstructions. Although it is generally assumed that such biases exist, little is known about their magnitude. To investigate this problem, we studied age and sex differences in the preservation of skeletal remains from Mission La Purisima and a prehistoric cemetery (Ca-Ven-110). Comparison of mortality profiles obtained through analysis of skeletal remains and burial records from the mission indicates that biases in preservation can be very significant in poorly preserved skeletal collections. The Purisima burial records show that most of the people interred in the cemetery were either infants or elderly adults. The skeletal remains, in contrast, are predominantly those of young adults. The burial records and skeletal collection produced comparable sex ratios. These results show that age biases in preservation are much more important than sex biases. This conclusion is supported by data on the completeness of the skeletons from La Purisima and Ca-Ven-110. At both sites, the remains of young adults were better preserved than those of children or elderly adults, and the completeness of male and female skeletons was comparable. PMID- 3046373 TI - Activity-induced patterns of dental abrasion in prehistoric Pakistan: evidence from Mehrgarh and Harappa. AB - A detailed investigation of worn teeth should reveal a record of past activity patterns including information regarding diet, food preparation methods, and craft or occupational activities. Anthropological studies of the extensive dental samples from Neolithic (MR 3) and Chalcolithic (MR 2) levels at Mehrgarh, Baluchistan, and Bronze Age Harappa, Punjab, yielded several interesting examples of unusual dental abrasion. This paper provides macro- and microscopic (scanning electron microscope) descriptions of three types of activity-induced dental abrasion: 1) interproximal tooth grooving and interproximal abrasion patches, 2) facial abrasion of maxillary anterior teeth, and 3) lingual abrasion of maxillary incisors in association with rounded wear of lower incisors. The gross size and shape of abrasion features, the orientation of microscopic wear striae, and ethnographic parallels are employed in inferring causal factors involved in their formation. Behavioral activities and dietary explanations possibly associated with each type of dental wear are considered and their implications for reconstructing prehistoric activity patterns discussed. The need for extensive ethno-anthropology research into variations of tooth use among living people with different diets, subsistence bases, and craft specializations is essential to further progress in this field. PMID- 3046372 TI - Brachydactyly, a possible inherited anomaly at prehistoric Prince Rupert Harbour. AB - Disproportionately short metacarpals or metatarsals in eight burial skeletons and three unusually short metapodials recovered as disturbed bones were identified in a 1500 B.C. to A.D. 500 skeletal series from eight archeological sites of the north mainland coast of British Columbia, Canada. At least ten people were affected from four sites for a minimum series frequency of 5.2%. Various factors clinically implicated in the occurrence of brachymetapody were investigated to account for the anomaly. Context-sensitive information suggested that trauma, infarction or infection, and individual or family-related malformation syndromes were unlikely possibilities. Some modern population data suggest that the series frequency was unusually high, particularly for fourth metatarsal involvement, the most commonly affected bone. Modern pedigree interpretations, ethnohistoric inferences, and the archeological contexts of the affected burial skeletons and site samples provide a framework for concluding that brachymetapody in the series was more likely due to the inheritance of an essentially isolated anomaly. PMID- 3046374 TI - Long-term hypotensive effect of beta-agonist in conscious dogs. AB - The purpose of this study was to investigate the arterial pressure response to long-term administration of beta-agonists in the chronically instrumented conscious animal model. Chronically instrumented dogs were given intravenous infusions of ritodrine (2 micrograms.kg-1.min-1) for a period of 2 wk. Several cardiovascular and renal parameters were monitored before, during, and after the ritodrine infusion, and renal function curves were constructed. After the 1st wk of infusion, a new steady state was reestablished, and this was characterized by hypotension, reduced plasma protein concentration, elevated cardiac output, expanded extracellular fluid space, and near normal levels of activity of renin angiotensin-aldosterone systems. The renal function curve during ritodrine infusion shifted to the left with no change in slope. We propose the following: 1) the persistence of hypotension is most probably related to the resetting of the arterial pressure-kidney blood volume servocontrol mechanisms, and 2) the persistent elevation of cardiac output and reduction in peripheral resistance are most probably related to increased oxygen and nutrient demand during beta-agonist infusions. PMID- 3046375 TI - Multiple linear regression is a useful alternative to traditional analyses of variance. AB - Physiologists often wish to compare the effects of several different treatments on a continuous variable of interest, which requires an analysis of variance. Analysis of variance, as presented in most statistics texts, generally requires that there be no missing data and often that each sample group be the same size. Unfortunately, this requirement is rarely satisfied, and investigators are confronted with the problem of how to analyze data that do not strictly fit the traditional analysis of variance paradigm. One can avoid these pitfalls by recasting the analysis of variance as a multiple linear regression problem. When there are no missing data, the results of a traditional analysis of variance and the corresponding multiple regression problem are identical; when the sample sizes are unequal or there are missing data, one can use a regression formulation to analyze data that cannot be easily handled in a traditional analysis of variance paradigm and thus overcome a practical computational limitation of traditional analysis of variance. In addition to overcoming practical limitations of traditional analysis of variance, the multiple linear regression approach is more efficient because in one run of a statistics routine, not only is the analysis of variance done but also one obtains estimates of the size of the treatment effects (as opposed to just an indication of whether such effects are present or not), and many of the pairwise multiple comparisons are done (they are equivalent to t tests for significance of the regression parameter estimates). Finally, interaction between the different treatment factors is easier to interpret than it is in traditional analysis of variance. PMID- 3046376 TI - Plasma dopamine in regulation of canine renal blood flow. AB - Studies were performed in pentobarbital-anesthetized dogs to determine whether circulating plasma dopamine (DA) is involved in renal blood flow (RBF) regulation. During graded reductions in renal perfusion pressure (RPP), total renal venous (RV) DA content significantly increased at RPPs below the autoregulatory range. The RBF response to decrements in RPP was also examined during control, infusion of DA (1.2 micrograms.kg(-1).min(-1)ia), and after DA receptor blockade by SCH 23390 (30 micrograms/kg iv). During DA infusion, autoregulation was still evident over the same RPPs, although at higher flow rates. At pressures below the autoregulatory range, RBF decreased linearly and the autoregulatory curve merged with control at 50 mmHg. After SCH 23390, autoregulation ceased at a higher RPP than during control, and RBF was significantly less than control rates at pressures of 80 mmHg and below. To elucidate reasons for this latter response, reductions in RPP were repeated before and after administration of both prazosin (0.1 mg/kg iv) and SCH 23390. The results indicated that RBF rates were not different from control at any RPP. Further, prazosin alone did not alter renal autoregulation but significantly increased RBF at RPP below the autoregulatory range. Thus these results indicate that dopamine does not participate in RBF control at pressures above the inflection point for the lowest limit of RBF autoregulation but may be released at lower RPP to act as a vasodilator agent to oppose alpha-adrenoceptor-mediated reductions in RBF. Moreover, tonic DA receptor activation may influence the setting of the lower limit of canine RBF autoregulation. PMID- 3046378 TI - Landfalls, journeys, and departures. PMID- 3046379 TI - George H. Pollock, M.D., Ph.D., one hundred sixteenth president, 1987-1988. American Psychiatric Association. PMID- 3046377 TI - Role of metabolic feedback regulation in glucose production of running rats. AB - Hepatic glucose production (Ra) was studied in phlorizin (P)- and saline (C) infused rats running for 35 min at 21 m/min in the postabsorptive state (series I) or 18 m/min in the fed state (series II). Phlorizin-induced increase in glucose clearance would increase or not affect Ra, depending on whether metabolic feedback mechanisms or central command from central nervous system (CNS), respectively, regulate Ra during exercise. Initial exercise-induced increases in Ra were similar in P and C rats of both series, although glucose clearance was higher and plasma glucose lower, however not hypoglycemic, in P rats. After 5 min of exercise, Ra remained similar in P and C rats in series I, whereas in series II, Ra increased almost twice as much in P compared with C rats. In both series muscle glycogenolysis and lipolysis were higher in P than in C rats. The results suggest a central command regulation of Ra from CNS motor centers and that this primary setting may be modulated by metabolic feedback mechanisms. Hypoglycemia is not needed to activate metabolic feedback. A variety of substrates rather than glucose specifically is mobilized by metabolic feedback mechanisms associated with decreased glucose availability. PMID- 3046380 TI - Conceptual and methodological issues in comparative studies of psychotherapy and pharmacotherapy, II: Nature and timing of treatment effects. AB - This is the second of two articles on the conceptual and methodological problems involved in comparing the effectiveness of drugs and psychotherapy in the treatment of mental disorders. Part II focuses on differences between psychotherapy and pharmacotherapy in the nature of treatment effects and related goals for treatment, differences in the time course of treatment effects, and potential sources of bias in the research setting. In designing comparative studies of psychotherapy and pharmacotherapy, investigators should address methodological choices explicitly and consider the implications for interpretation of findings. PMID- 3046381 TI - Lack of efficacy of carbamazepine in the treatment of panic disorder. AB - The authors conducted a controlled study of carbamazepine in the treatment of 14 patients with panic disorder. Although there was a statistically significant reduction in symptoms of anxiety on several measures, only one of the patients was judged to have a marked and sustained clinical improvement while taking carbamazepine. Forty percent of the patients had a decrease in frequency of panic attacks during carbamazepine treatment, 50% had an increase, and 10% showed no change. The presence of either EEG abnormalities or prominent psychosensory symptoms did not predict response to carbamazepine. These findings are discussed within the context of an epileptiform model for panic disorder. PMID- 3046382 TI - S-adenosylmethionine treatment of depression: a controlled clinical trial. AB - The antidepressant properties of S-adenosylmethionine, an endogenous methyl donor, were studied in inpatients who met the DSM-III criteria for major depression. Nine patients given intravenous S-adenosylmethionine and nine given low oral doses of imipramine were compared in a double-blind design for 14 days. The S-adenosylmethionine produced superior results by the end of the first week of treatment. By the end of the second week, 66% of the S-adenosylmethionine patients had a clinically significant improvement in depressive symptoms, compared to 22% of the imipramine patients. Side effects appeared to be fewer with S-adenosylmethionine than with imipramine during the last 5 days of the study. PMID- 3046384 TI - Placebo response in panic disorder. AB - Of 43 patients with panic disorder or agoraphobia with panic attacks who took placebo for 8 weeks in two double-blind studies, one in four markedly improved. Those with consistently normal dexamethasone suppression test results were significantly more likely to show a placebo response as were those with lower anxiety ratings at the outset of treatment. PMID- 3046383 TI - A randomized clinical trial of inpatient family intervention, III: Effects at 6 month and 18-month follow-ups. AB - This paper focuses on the follow-up results of a randomized clinical trial of inpatient family intervention (IFI) that emphasized psychoeducation. Results for the sample of 169 psychiatric patients suggested that adding family treatment to standard hospital treatment was effective; however, the statistical interactions indicated that this therapeutic effect was restricted to female patients with schizophrenia or major affective disorder. The effect of family treatment on male patients with these diagnoses was minimal or slightly negative. In a group of patients with other diagnoses, the Treatment by Sex effect was reversed: male patients did better with the family treatment. PMID- 3046385 TI - DST results in nonpsychotic depressed outpatients. AB - In two studies using the dexamethasone suppression test (DST) to evaluate the efficacy of newer antidepressants in depressed outpatients, the authors found a DST nonsuppression rate of 13% (11 of 86 patients). Thirty-three of the DST suppressors received an antidepressant and 42 received placebo; the drug-treated group showed a significant therapeutic response. The low rate of DST nonsuppression in these depressed outpatients, a finding consistent with that of other investigators, does not confirm or refute reports that these patients are relatively resistant to placebo in comparison with active medication. The authors recommend that DST results not be used as selection criteria in studies assessing newer therapies for depressed outpatients. PMID- 3046386 TI - Fluoxetine treatment of obsessions and compulsions in patients with Tourette's syndrome. PMID- 3046388 TI - Prophylactic knee bracing. AB - Six studies to determine the effectiveness of prophylactic knee braces in preventing medial collateral ligament (MCL) injury in football are compared. Criteria useful in evaluating studies are discussed, such as the probability of confounding factors, bias in selecting cases and controls, and variations in defining injury and exposure. Cost, as well as some ethical issues associated with mandated use are discussed. While four of these studies found a reduction in MCL injuries associated with using a brace, two of them reported increases. No consensus arises from these studies, conflicting results as well as methodological problems in some make it impossible to state with assurance the role of prophylactic knee bracing in football at this time. PMID- 3046387 TI - The medical aspects of soccer injury epidemiology. AB - In this article, the six major studies of soccer injury epidemiology are reviewed. Strengths and weaknesses of each epidemiologic design are critiqued and the crucial importance of the definition of injury is emphasized. The effect of age, sex, and intensity of play on injury rates is discussed. Our present knowledge of injury rate by anatomical site, player position, and the type of playing surface are reviewed. We examined the importance of player factors such as flexibility, joint laxity, weakness, and incomplete rehabilitation from other injuries. In addition, we reviewed the role played by inadequate equipment, field conditions, and rule violations. A successful program for soccer injury prevention is described, and guidelines for future soccer injury epidemiology research are proposed. PMID- 3046389 TI - Injuries in women's gymnastics. The state of the art. AB - This article takes a three-stage approach to the topic of injuries in women's gymnastics. In the first stage, we review the literature and summarize our current knowledge of injury rates, injuries in specific events, and anatomical location of injuries. In the second, we critically evaluate the relative contributions of these studies in terms of their generalizability, methods, and conceptual approaches. Finally, we present possible directions for future research in the area of women's gymnastic injuries. PMID- 3046390 TI - Sports injury research. How to choose a study design. AB - In conducting a clinical study, one must determine the timing of the study. A cohort study may be either retrospective or prospective. A case-control study is retrospective and compares subjects with a particular condition to controls without that condition by evaluating events which preceded the development of the condition. A cross-sectional study examines subjects at one particular moment in time. Once the timing of a study is determined, the type of control group is selected. One may present an uncontrolled series or use historical or concurrent controls. The prospective, concurrently controlled series is the strongest of the above designs. The randomized clinical trial is the strongest study design and is employed to avoid the errors that are potentially present in non-randomized studies (Chapter 9). It is a prospective, concurrently controlled study design in which subjects are randomly allocated to two or more treatment modalities. The protocol specifies the population source, admission criteria, enactment of the procedure, and outcome determination for the study. PMID- 3046391 TI - Sports injury research. How to avoid bias. AB - A valid study is one that is accurate and without bias. Bias may occur in a study in four major ways. Susceptibility bias may arise if groups are not similar at the initial state. The potential for performance bias exists if procedures are not performed comparably. Detection bias may occur if outcomes are not measured comparably. Transfer bias occurs if there is differential loss at followup. The major advantage of a randomized clinical trial is in its attempt to prevent susceptibility bias. Despite the methodologically correct implementation of a study, however, results may not be generalizable due to the particular population, restrictive eligibility criteria, or poor participation. An investigator must balance dual goals: conducting a valid trial and providing generalizable results. PMID- 3046392 TI - Ice hockey injuries. AB - Ice hockey is a team sport that has recently grown in popularity not only in the United States but also in Canada and Europe. With this increase in popularity has come a growing concern about the number and severity of injuries. The world literature on the biomechanics and physiology of ice hockey was reviewed in an attempt to evaluate the forces and mechanisms involved in the game. The influence of rule and equipment changes on injury patterns was particularly studied. Several studies on the epidemiology of injuries, providing data on the types of injuries and the mechanisms of those injuries, were analyzed to determine the conclusions that could be supported and those that require further study. Possible changes in the patterns and types of injury are outlined. PMID- 3046394 TI - Granulomatous inflammation in pineal germinoma. A cause of diagnostic failure at stereotaxic brain biopsy. AB - We report four patients with pineal germinoma in whom the initial procedure for obtaining a tissue diagnosis was a stereotaxic biopsy. In all four cases, the biopsy showed granulomatous inflammation with epithelioid cells and lymphocytes. In one case, the granulomatous inflammation was accompanied by classical germinoma. Another case exhibited malignant cells considered nondiagnostic for a specific neoplasm. Two cases had no evidence of a malignant tumor; they were composed entirely of granulomatous inflammation. In two of the three cases where the diagnosis was not established by the first biopsy, a second stereotaxic biopsy showed pineal germinoma. Tissue obtained at resection in the third patient revealed large areas of granulomatous inflammation accompanying the germinoma. Immunoperoxidase stains for ferritin and placental alkaline phosphatase did not increase diagnostic yield. We conclude that a finding of granulomatous inflammation in a stereotaxic biopsy specimen of a pineal mass should suggest a diagnosis of germinoma, followed by sampling from several different target points within the lesion. PMID- 3046393 TI - High school football injuries: evaluation. AB - An epidemiologic survey of the literature on high school football injuries revealed methodologic problems. These numerator-denominator inconsistencies and other confounding factors are discussed. The authors suggest a more reliable system of reporting these parameters to further reduce the risk of high school football injuries. PMID- 3046395 TI - Immunohistochemical detection of epithelial membrane antigen in normal perineurial cells and perineurioma. AB - The use of epithelial membrane antigen (EMA) as an immunohistochemical marker for normal and neoplastic perineurial cells is described. Normal perineurial cells react strongly for this antigen, which is also expressed by the cells of perineurioma. Instead, neurofibromas and schwannomas only show some peripheral or entrapped layers of EMA-positive cells. In traumatic and Morton's neuromas, bundles of neural fibers are wrapped in layers of EMA-positive perineurial cells. Neurothekeoma and granular cell tumor show no EMA reactivity. The detection of an epithelial marker in perineurial cells is in agreement with the concept of a "perineural epithelium" and seems to support a common embryologic origin for the perineurial cell and the equally EMA-positive arachnoidal cap cell. The availability of an immunohistochemical marker for the perineurial cell provides an easy and convenient tool for the evaluation of the participation of this cell in a variety of pathologic processes. PMID- 3046396 TI - Bile duct adenoma. A study of 152 cases. AB - In order to determine the morphologic spectrum of bile duct adenoma (BDA), we reviewed the clinical, gross, and histopathological features of 152 cases. All BDA were asymptomatic nodules discovered incidentally during intra-abdominal surgery (103 cases) or at autopsy (49 cases). They were usually subcapsular, ranged in size from 1 to 20 mm (mean, 5.8 mm), and were well circumscribed but nonencapsulated. Histologically, BDA was composed of benign, noncystic ductules and variable degrees of inflammation and fibrosis. The immunophenotype of these ductules was similar to that of interlobular bile ducts. Follow-up of 38 of the surgically treated patients confirmed the benign behavior of this lesion. BDA should be distinguished from an adenocarcinoma by the absence, in the former, of nuclear hyperchromasia, mitotic activity, and vascular invasion. The absence of bile and cystic changes and lack of association with polycystic disease of the liver and kidneys are the main features distinguishing BDA from von Meyenburg complex. We believe that BDA is a reactive process to a focal injury rather than a true neoplasm or a developmental anomaly. PMID- 3046397 TI - [Trends and factors in maternal mortality throughout the world]. PMID- 3046398 TI - [Immunomodulating prophylaxis via skin grafts in pregnant women at risk for the development of late toxicoses]. PMID- 3046399 TI - [Comparative double-blind study of intravenous metronidazole versus placebo in preventing infection after cesarean section]. PMID- 3046400 TI - [Acquired immunodeficiency syndrome (AIDS) and its relation to obstetrics and gynecology]. PMID- 3046401 TI - [Prof. Nikolai Zhekov Markovski]. PMID- 3046402 TI - Cutaneous plasmacytic infiltrates. AB - This article reviews a group of skin disorders that have in common the presence of a cellular infiltrate with plasma cells upon histopathologic examination. These include inflammatory and neoplastic diseases and a group of miscellaneous disorders of varied etiology. The correlation between the presence of plasma cells in the cellular infiltrate and the diagnosis, prognosis, and pathogenesis of the disorders is emphasized. PMID- 3046403 TI - Morris Leider, M.D. PMID- 3046404 TI - Cumulative dose response relationship of terbutaline delivered by three different inhalers. AB - In a controlled, open cross-over study 15 asthmatic children received increasing doses of terbutaline (0.125 mg + 0.125 mg + 0.25 mg + 0.5 mg + 0.1 mg) delivered by a pressurized aerosol alone or with a tube spacer or a nebuhaler attached. At no time was there any significant difference in measured FEV1 or percent increase in FEV1 between the three inhalers. However, when eight patients with bronchodilation greater than 50% on all tests days were studied separately, a significantly greater improvement in FEV1 was seen after the first three inhalations of terbutaline when a nebuhaler or a tube spacer was used compared with the response after the pressurized aerosol (P less than 0.05) Bronchodilation measurements did not differ after use of the two spacers. There was no statistically significant difference in side effects between the three inhalers, but there was a trend towards a higher occurrence of side effects when the pressurized aerosol was used alone. PMID- 3046405 TI - Immunotherapy with partially purified and standardized tree pollen extracts. I. Clinical results from a three-year double-blind study of patients treated with pollen extracts either of birch or combinations of alder, birch and hazel. AB - Fifty-four adult patients with tree pollen-induced rhinitis (28), asthma (1), or rhinitis and asthma (25) were selected for immunotherapy with standardized and partly purified tree pollen extracts using a double blind protocol. The selection was based on clinical history, results of nasal or bronchial challenge, skin prick tests and RAST. Further, based on crossed radio-immunoelectrophoresis, sex, age and severity of symptoms, the patients were allocated in matched pairs and the treatment alternatives were randomly distributed within the pairs. Twenty three patients treated with extracts composed of any combination of alder, birch and hazel pollen which matched their IgE response in CRIE (Group 1 (ABC)) and 22 patients receiving birch pollen extracts (Group 2 (B)) completed all 3 years of treatment. The in vivo results comprising symptom and medicine consumption scores are given here. Changes in specific skin and nasal reactivity as well as in immunological parameters are presented separately. No significant differences were demonstrated between the treatment groups in the two parameters. Both extracts were effective and reduced in general the symptom scores to one tenth of the starting level. Expressed another way, at the end of the study, the patients tolerated 30 times more pollen until symptoms of the same severity were elicited, compared to before. In the Nordic countries, spring-time asthma and rhino conjunctivitis caused by pollen from deciduous trees can be effectively treated with an extract of birch pollen alone. PMID- 3046406 TI - Immunotherapy with partially purified and standardized tree pollen extracts. II. Results of skin prick tests and nasal provocation tests from a three-year double blind study of patients treated with pollen extracts either of birch or combinations of alder, birch and hazel. AB - Patients allergic to tree pollen entered a 3-year course of immunotherapy (1980 83) with either birch pollen extracts alone (n = 26) or patient-tailored extracts of birch, alder and hazel pollen (n = 27). The clinical and immunological results of this study are published elsewhere. This paper contains an evaluation of skin prick test and nasal provocation test results. There were no significant differences between the two treatment groups concerning these two parameters. In both groups the allergen-specific sensitivity in the skin showed seasonal variations but a significant decrease. During the years of treatment there was also a significant decrease in the specific sensitivity of the nasal mucosa. With the present demands for purification and standardization of allergen extracts it is of practical and economic interest to know that tree pollen-allergic patients showing positive reactions to birch, alder and hazel extracts can be effectively treated using birch pollen extract alone. PMID- 3046407 TI - Immunotherapy with partially purified and standardized tree pollen extracts. III. Specific IgE response to the major allergens of alder, birch and hazel pollen during immunotherapy. AB - Patients allergic to pollen from alder, birch and hazel were hyposensitized during a 3-year period with either birch pollen extract alone (n = 24) or a mixture of one or more of alder, birch and hazel pollen extracts (n = 27). The effect of the treatment was evaluated by RAST and tandem crossed radioimmunoelectrophoresis (tandem-CRIE). The patient' specific IgE response to the major allergens of alder (Aln g I), birch (Bet v I) and hazel (Cor a I and Cor a II), as measured by tandem-CRIE, and the total specific IgE response, measured by RAST, decreased significantly (Pc less than 0.05) during immunotherapy, irrespective of the extract used during the treatment. There was no significant difference (Pc less than 0.05) between the two treatment groups. The results obtained indicate either that birch pollen extract alone is adequate in the treatment of the studied patient group or the patients had been sensitized towards birch pollen alone. PMID- 3046409 TI - Turbuhaler: a new device for dry powder terbutaline inhalation. AB - The aim of this study was to compare bronchodilator response and adverse effects of terbutaline when administered with the metered dose inhaler (MDI) Bricanyl and with the dry powder inhaler Bricanyl Turbuhaler (BT). Nine adult patients with bronchial asthma participated. The study was of an open crossover design. At 30 min intervals the patients inhaled increasing doses of terbutaline (0.25 mg to 5.0 mg cumulated) from either the conventional MDI or from the BT. After each inhalation FEV1, FVC, heart rate, muscle tremor and adverse effects were recorded. Both treatments, BT and MDI, resulted in a dose-related increase in lung function, without any statistical difference. Taste sensation, muscle tremor and increase in heart rate were observed in both groups. Because of the design of the BT one may assume that inhalation failure can be avoided. PMID- 3046408 TI - Effect of topical levocabastine on allergic and non-allergic perennial rhinitis. A double-blind study, levocabastine vs. placebo, followed by an open, prospective, single-blind study on beclomethasone. AB - Forty-four patients, with symptoms of nasal obstruction, sneezing, itching and/or rhinorrhea, were entered into a placebo-controlled, double-blind study to evaluate the clinical efficacy of a topical antihistamine drug, levocabastine, applied 4 times a day for 14 days. At the end of the treatment the placebo patients were treated with levocabastine and the levocabastine patients were treated with beclomethasone dipropionate in a single-blind design for another 14 days. This study showed that levocabastine is significantly more active than placebo with reference to nasal discharge and sneezing. Placebo application improved the symptom score. Levocabastine could not be proved to be more effective against nasal obstruction than placebo in the double-blind trial. In the single-blind set-up, levocabastine resulted in an additional improvement in the score for obstruction, after the placebo period. Although the allergic group tended to respond better, no statistically significant difference could be detected between allergic and non-allergic patients. After treatment with levocabastine, beclomethasone dipropionate administration could not improve the results for nasal discharge and sneezing. For nasal congestion, beclomethasone dipropionate proved to be superior to levocabastine. PMID- 3046410 TI - Diabetes and anaesthesia--slow progress? PMID- 3046411 TI - Insulin treatment of the insulin-dependent diabetic patient undergoing minor surgery. Continuous intravenous infusion compared with subcutaneous administration. AB - In a prospective randomised study in 20 insulin-dependent diabetics who had minor surgery under general anaesthesia we compared the metabolic responses to intravenous glucose-insulin-potassium infusion with those who had conventional subcutaneous insulin administration. The former treatment resulted in lower blood glucose levels both during the infusion period (p less than 0.05) as well as the entire observation period (operative, first and second postoperative days; p less than 0.01). More blood glucose values were within the intended range of 5 to 10 mmol/litre in the glucose-insulin-potassium as compared to the conventional group (48% versus 24%; p less than 0.01). The levels of lactate, 3-hydroxybutyrate, glycerol, alanine, glucagon, insulin and growth hormone did not differ between the two groups. The infusion regimen resulted in better glycaemic control both peri-and postoperatively than the conventional subcutaneous insulin regimen in insulin-dependent diabetic patients who have minor surgery. PMID- 3046413 TI - [Amrinone (Wincoram)--a new positive inotropic and vasodilator agent]. AB - The bipyridine derivative amrinone is a specific phosphodiesterase III blocking agent. In vitro and in vivo studies show a dose-dependent increase in myocardial contractility induced by amrinone. In patients with congestive heart failure, the inotropic and vasodilator effects of amrinone contribute to cardiac improvement. When amrinone is used, the increase in myocardial oxygen consumption due to increased contractility is offset by the reductions in preload and afterload. In hearts with very high wall tension, myocardial oxygen consumption may even decrease with amrinone. Amrinone therapy is not accompanied by significant increases in heart rate. Tachyphylaxis has not been observed. The elimination half-life ranges between 2.5 and 3.5 h. A large quantity of amrinone is excreted unchanged, and therefore in cases of renal impairment the possibility of cumulation exists. The main adverse reaction of amrinone is a reversible thrombocytopenia induced by a dose-dependent decrease in platelet survival time. Therefore, frequent platelet counts are necessary when amrinone is administered. Numerous studies in patients with chronic congestive heart failure confirmed the beneficial hemodynamic effects of amrinone. Experience in the treatment of acute perioperative heart failure with amrinone are still limited, but the present results are encouraging; an additive effect of amrinone to catecholamines seems especially promising in the therapy of severe postoperative low-cardia-output syndrome. PMID- 3046412 TI - Diabetes mellitus and anaesthesia. A survey of the peri-operative management of the patient with diabetes mellitus. AB - A variety of methods are currently available for the management of the diabetic patient in the peri-operative period. A questionnaire about current clinical practice was sent to all anaesthetists in the Oxford region. The majority reported that minor surgery in both insulin treated and noninsulin treated diabetic patients warranted no intervention other than avoidance of meals and medication before surgery, and that, for major surgery, a glucose-insulin potassium infusion should be used. Fifty one out of 71 respondents in the junior staff grades preferred this latter approach for intermediate surgical procedures in insulin treated patients compared with 27 out of 69 of the consultant staff. Most anaesthetists aimed for blood glucose levels of 7-13 mmol/litre in the peri operative period. The literature is also reviewed. PMID- 3046415 TI - Nuclear magnetic resonance spectroscopy. PMID- 3046414 TI - [The agonist-antagonist nalbuphine prolongs gastro-cecal transit time and induces short-term pain following neuroleptanesthesia using fentanyl. A comparative study using a placebo]. AB - Little is known about the effects of mixed opioid analgesics on gastrointestinal propulsion. In 20 patients, nalbuphine (0.1 mg/kg) was given after routine neuroleptanesthesia consisting of 70 micrograms/kg droperidol, 7 micrograms/kg fentanyl, and N2O/O2 (3:1) ventilation, to study its effect on gastrointestinal motility in the postoperative period. For comparison, another group of patients (n = 20) undergoing similar interventions received placebo (0.9% NaCl) at the end of the procedure. Gastrointestinal transit time was determined by measuring the exhaled H2 concentration following gastric lactulose administration. As lactulose is degraded only in the cecum, resulting in the release of hydrogen, the arrival of the polysaccharide at the terminal ileum could thus be determined. Compared to placebo, gastrointestinal transit was significantly longer in patients after nalbuphine (mean transit time 270 min vs 380 min). Pain estimation by visual analogue scale (VAS 0-10) suggested an antagonistic effect at the 10th and 20th min postoperatively, as pain scores in the nalbuphine group were higher when compared to placebo (3.5 vs 1.8 and 2.5 vs 1.4). There was a similar pain score in both groups (1.3 vs 1.4) 30 min after drug administration. However, there was significantly better pain relief after nalbuphine (0.7 vs 1.4 and 0.7 vs 1.1) in the late postoperative period (120th and 240th min). When given after potent opioids, it must be borne in mind that the antagonistic effect of nalbuphine is initially apparent. The agonistic potency of the compound will come into effect around the 30th min post-injection. Delayed gastrointestinal transit after nalbuphine is explained by agonist-like effects on peripheral opioid receptors in the gut. PMID- 3046417 TI - Atomic mass spectrometry. PMID- 3046416 TI - Ion-selective electrodes. PMID- 3046418 TI - Ultraviolet and light absorption spectrometry. PMID- 3046419 TI - Dynamic electrochemistry: methodology and application. PMID- 3046421 TI - Atomic absorption, atomic fluorescence, and flame emission spectrometry. PMID- 3046420 TI - Molecular fluorescence, phosphorescence, and chemiluminescence spectrometry. PMID- 3046422 TI - Thermal analysis. PMID- 3046423 TI - Gas chromatography. PMID- 3046424 TI - X-ray spectrometry. PMID- 3046425 TI - Mass spectrometry. PMID- 3046426 TI - Emission spectrometry. PMID- 3046428 TI - Surface analysis: X-ray photoelectron spectroscopy and Auger electron spectroscopy. PMID- 3046427 TI - Raman spectroscopy. PMID- 3046429 TI - Column liquid chromatography. PMID- 3046431 TI - Thin-layer and paper chromatography. PMID- 3046430 TI - Chemical sensors. PMID- 3046432 TI - The new electron microscopy: imaging the chemistry of nature. PMID- 3046433 TI - Mossbauer spectroscopy. PMID- 3046436 TI - Migration of an arterial catheter. PMID- 3046435 TI - Neutrophil lactoferrin content: variation among mammals. AB - Lactoferrin (Lf) in blood and/or marrow neutrophils was semiquantified using indirect immunofluorescence technique in nine mammalian species. Neutrophil iron binding reactivity (NFeBR), which corresponds primarily to Lf, was also visualized and semiquantified using functional cytochemical (FeNTA-AF) technique at the light microscopic level in these nine and in an additional fifteen mammalian species, and in selected species at the ultrastructural level. Neutrophil immunoreactive Lf was positively correlated with total cellular and granule content of NFeBR among these nine species, and with previously reported concentrations of neutrophil Lf quantified by radioimmunoassay. Relative levels of Lf in neutrophil extracts from rat, hamster, and human were confirmed using SDS-polyacrylamide gel electrophoresis and immunoblotting. Relatively high levels of immunoreactive neutrophil Lf and/or NFeBR were observed in carnivores (ten species) and primates (six species). Among rodents (five species), the levels were variable, and the artiodactyls (four species) studied had low levels. These results demonstrate that neutrophil Lf levels vary widely among mammalian species. In addition, FeNTA-AF technique provides a rapid means of evaluating animals for relative quantities of neutrophil Lf. PMID- 3046434 TI - Distribution of intermediate filament proteins in developing and adult salivary glands in man. AB - Adult and developing salivary glands were investigated using five monoclonal antibodies against cytokeratins (CKs) and vimentin. Acinar cells displayed mainly CK 18 whereas CKs 7, 17 and 19 were only detected in duct and myoepithelial cells. All epithelial and myoepithelial cells were unreactive for one vimentin antibody (Vim 9) whereas with the other (Vim 24), myoepithelial cells and basal cells of excretory ducts were stained. Fetal cells showed the CK pattern of duct cells. At gestational week 18, a reaction for both vimentin antibodies could be found in basal cells of terminal tubules. Although vim 9 reactivity has been shown for a number of salivary neoplasms, it has not been detected in any adult epithelial salivary tissue. The finding of this reactivity in the fetal gland indicates that the expression of this intermediate filament protein in certain salivary neoplasms may be a sign of dedifferentiation resulting in the expression of a filament pattern found in an earlier stage of gland development. PMID- 3046437 TI - Anesthesia with halothane and nitrous oxide alters protein and amino acid metabolism in dogs. AB - General anesthesia in combination with surgery is known to result in negative nitrogen balance. To determine whether general anesthesia without concomitant surgery decreases whole body protein synthesis and/or increases whole body protein breakdown, two groups of dogs were studied: Group 1 (n = 6) in the conscious state and Group 2 (n = 8) during general anesthesia employing halothane (1.5 MAC) in 50% nitrous oxide and oxygen. Changes in protein metabolism were estimated by isotope dilution techniques employing simultaneous infusions of [4,53H]leucine and alpha-[1-14C]-ketoisocaproate (KIC). Total leucine carbon flux was unchanged or slightly increased in the anesthetized animals when compared to the conscious controls, indicating only a slight increase in the rate of proteolysis. However, leucine oxidation was increased (P less than 0.001) by more than 80% in the anesthetized animals when compared with their conscious controls, whereas whole body nonoxidative leucine disappearance, an indicator of whole body protein synthesis, was decreased. The ratio of leucine oxidation to the nonoxidative rate of leucine disappearance, which provides an index of the catabolism of at least one essential amino acid in the postabsorptive state, was more than twofold increased (P less than 0.001) in the anesthetized animals regardless of the tracer employed. These studies suggest that the administration of anesthesia alone, without concomitant surgery, is associated with a decreased rate of whole body protein synthesis and increased leucine oxidation, resulting in increased leucine and protein catabolism, which may be underlying or initiating some of the protein wasting known to occur in patients undergoing surgery. PMID- 3046438 TI - Independent lung ventilation using high-frequency ventilation in the management of a bronchopleural fistula. PMID- 3046440 TI - Principles of pharmacotherapy: I. Pharmacodynamics. AB - This paper and the ensuing series present the principles guiding and affecting the ability of drugs to produce therapeutic benefit or untoward harm. The principles of pharmacodynamics and pharmacokinetics, the physiologic basis of adverse drug reactions and suitable antidotal therapy, and the biologic basis of drug allergy, drug-drug interactions, pharmacogenetics, teratology and hematologic reactions to chemicals are explored. These principles serve to guide those administering and using drugs to attain the maximum benefit and least attendant harm from their use. Such is the goal of rational therapeutics. PMID- 3046441 TI - Principles of pharmacotherapy: II. Pharmacokinetics. PMID- 3046439 TI - Cardiac dysrhythmias with general anesthesia during dental surgery. AB - Dysrhythmias with general anesthesia during dental surgery have been frequently reported. The incidence appears higher in spontaneously breathing patients lightly anesthetized with halothane. Anxiety, sitting posture, hypoxia, Chinese race, and heart disease appear to aggravate the condition. Use of beta blockers or lidocaine prior to anesthesia, intravenous induction, controlled ventilation with muscle relaxants, and use of isoflurane or enflurane in spontaneously breathing patients appear to decrease the incidence. It is stressed that continuous cardiac monitoring should be done in patients undergoing dental surgery under anesthesia in order to detect diagnose and treat any dysrhythmia. The great majority of dysrhythmias disappear either spontaneously or when the stimulus is stopped. In some cases there may be an obvious cause that should be immediately corrected. The need for drug intervention is rare and must be used with great care when used. PMID- 3046443 TI - [The production of hybrid cells producing monoclonal antibodies against Toxoplasma gondii]. AB - Hybridomas that secrete monoclonal antibody to Toxoplasma gondii have been developed. Two groups of 10 female BALB/c mice each were immunized either over a shorter (71 d) or longer (117 d) period at first with Toxoplasma lysate antigen and afterwards with intact tachyzoites of the RH strain. Higher titres of antibody were obtained with the long-period immunization. The fusion experiments have shown that both schemes of immunization approximately result in the same number (16 and 14% respectively) of hybridoma cell lines producing antigen specific monoclonal antibodies. Hybridoma cultures secreting antitoxoplasma monoclonal antibodies were screened parallel by indirect immunofluorescence antibody test (IFAT) and enzyme immunoassay (EIA). 16 of the hybridoma cell cultures produced positive results only in the IFAT, 112 reacted only in the EIA and 21 were positive in both tests. The monoclonal antibodies 5B10, 5G6 and 1B2, which are positive in the IFAT form a chemical compound with the major antigens on the surface of RH tachyzoites. The patterns of fluorescence produced by these monoclonal antibodies are in conformity with those produced by using polyclonal sera of Toxoplasma gondii infected hosts (mouse, rabbit, man). PMID- 3046442 TI - Propofol as an intravenous agent in general anesthesia and conscious sedation. AB - Propofol has been shown in clinical studies to be a safe, effective, hypnotic, and amnesic anesthetic agent at induction doses of 2-2.5 mg/kg and maintenance doses of approximately 9mg/kg per hour. Significant post-induction hypotension reported earlier can be reduced to a all in MAP of less than 25% when the drug is used alone (without nitrous oxide or narcotic premedication). Post-induction apnea is minimized by avoidance of pre-induction hyperventilation. Acute and long term venous tolerance is acceptable. Emergence from anesthesia induced and maintained with propofol is rapid, predictable and relatively free of postoperative complications. Incidence of drug interaction is low. Propofol causes no adrenocortical suppression and is not potentiated by ethanol, diazepam, amitriptyline or phenelzine. Preliminary investigation of propofol as an intravenous sedative agent at subanesthetic doses has been favorable. PMID- 3046444 TI - The approach to chronic cough in childhood. AB - Chronic cough is a fairly common pediatric complaint. Usually, it is secondary to irritation of the airways following a respiratory viral infection. In these cases, the cough tends to diminish over time. There may, however, be a subsequent development of bronchial hyperreactivity. Asthma is common in the pediatric population. From 50% to 90% of chronic coughers may have hyperreactive airways. In the absence of a pulmonary function laboratory to test for this, a trial of bronchodilator therapy is warranted. Other conditions discussed may also cause chronic cough and a thorough history and physical examination with some simple radiologic investigations can help pinpoint the cause. Specific therapy can then be used to manage the problem. In addition to specific therapy, care must be taken to explain to the parents and patient the physiology of the cough and why it is present. Anxieties and fears should be dealt with in a caring and direct manner. Occasionally, non-specific therapy is needed to allow the parents and child some rest and relief. PMID- 3046445 TI - Bronchial reactivity pattern in nonasthmatic parents of asthmatics. AB - Genetic mechanisms have been proposed to explain the presence of asthma in families. A methacholine challenge can identify individuals with bronchial reactivity (a hallmark of asthma). It may then be possible to determine whether the presence of non-specific bronchial reactivity, as detected by a methacholine response, has potential as a genetic marker. Thirty-one non-asthmatic parent pairs of asthmatic children were selected from asthma (AF) families enrolled in a Natural History of Asthma study. Parent pairs were chosen if both gave negative responses to a modified National Heart, Lung and Blood Institute questionnaire on asthma. The methacholine response of these parents of asthmatic children had a bimodal distribution. These results show that the methacholine response can mark bronchial reactivity without the presence of clinical asthma and that a familial component of bronchial reactivity exists which may be transmitted from one generation to the next. PMID- 3046446 TI - Post-traumatic neck pain: a prospective and follow-up study. AB - Three hundred fifty-one alert emergency department patients with post-traumatic neck pain were evaluated prospectively. Seven (2%) had proven fractures or ligament disruptions. The immediate onset of neck pain and the presence of posterior midline cervical tenderness each had 100% sensitivity, with specificity of 65% and 48%, respectively. Discharged patients were followed up by telephone or letter at a mean of 25 +/- 20 weeks. Of this group, 63% saw another physician, and 43% had persistent moderate-to-severe neck pain or neurologic symptoms at a mean follow-up time of 24 weeks after injury. Of those who had not had a cervical radiograph while in the ED, 52% later obtained one. In addition, 66% of the discharged patients were pursuing litigation. The results suggest that it may be possible to identify alert patients with cervical spine injury by means of history and examination only; however, a larger study confirming these results is required. The high frequency of post-ED radiography, the high prevalence of persistent symptoms after injury, and frequent involvement in litigation are factors to be considered when evaluating ED patients with post-traumatic neck pain. These factors may support the validity of obtaining cervical spine radiographs on many more of these patients than high-yield criteria would dictate. PMID- 3046447 TI - North American tick-borne diseases. PMID- 3046448 TI - Procurement and workbench procedures in preparation of pancreatic allografts. Factors essential for a successful pancreas transplant. AB - The rapidly developing field of pancreas allotransplantation can be regarded as an excellent school of surgery in which the topics of meticulous surgical technique and attention to detail by virtue of their significance strongly outweigh all other topics. The growing experience of the rather grave complications after pancreas transplantation points to the need for consistent use of meticulous techniques during organ procurement and application of workbench procedures just prior to transplantation. In addition, with the further development of the extra-renal transplant programs, the procurement of multiple organs from the same donor is becoming more common and the pancreatic team is more frequently faced with a less than optimal situation in regard to the vascular supply of the pancreatic graft. It, therefore, becomes imperative to carefully evaluate the vasculature and to make the necessary modifications prior to transplantation, since the best surgical techniques will not outweigh the consequences of a suboptimal blood supply, which will compromise the viability of any segment of the delicate pancreaticoduodenal graft. PMID- 3046450 TI - The reversible dementias: do they reverse? AB - Thirty-two studies (2889 subjects) that investigated the prevalence of the causes of dementia were critically reviewed. Particular attention was paid to potential and actual reversibility. Although dementia manifests itself primarily in old age (particularly age 75 and older), the mean age of patients for the studies that reported age data (56%) was 72.3 years. Twenty-five studies originated from secondary or tertiary centers, and four were community-based. Dementias consisted of Alzheimer disease, 56.8%; multi-infarct, 13.3%; depression, 4.5%; alcoholic, 4.2%; and drugs, 1.5%. No single other cause contributed more than 1.6% of the cases. Potentially reversible causes made up 13.2% of all cases. However, the more important question of whether patients with potentially reversible causes were followed and reversal actually seen was not always examined. In 11 studies (34%) that provided follow-up, 11% of dementias resolved, either partially (8%) or fully (3%). The commonest reversible causes were drugs, 28.2%; depression, 26.2%; and metabolic, 15.5%. Due to the presence of various biases (selection, lack of "blinded" investigators, and others) in the surveyed works, it is probable that the true incidence of reversible dementias in the community is even lower than that reported. Research implications as well as a conservative approach to the workup of a new case of dementia are offered. PMID- 3046449 TI - Functional asplenia after bone marrow transplantation. A late complication related to extensive chronic graft-versus-host disease. AB - STUDY OBJECTIVE: To evaluate splenic function in bone marrow transplant recipients, with relation to chronic graft-versus-host disease and infections. DESIGN: Survey, outpatients geographically accessible for voluntary participation. SETTING: Bone marrow transplantation referral center. PATIENTS: Fifteen bone marrow graft recipients (13 allogeneic, 2 autologous), out of a total of 33 patients who received transplants at the center and survived more than 6 months after grafting. MEASUREMENTS AND MAIN RESULTS: In 6 of 15 patients impaired splenic function (functional asplenia) was indicated by the presence of Howell-Jolly bodies in peripheral blood smears, reduced spleen size (P less than 0.001), higher platelet counts (P less than 0.01), higher indium-111 labeled autologous platelet recovery (P less than 0.005), reduced splenic blood flow (P less than 0.001), and reduced accumulation of radioactivity at the splenic site (P less than 0.001). All patients with functional asplenia but only 2 patients without functional asplenia had extensive chronic graft-versus-host disease. The incidence of bacterial infections was four times higher in patients with impaired splenic function. CONCLUSIONS: Functional asplenia is a late complication after allogeneic bone marrow transplantation and contributes to the high susceptibility to bacterial infections in patients with extensive chronic graft-versus-host disease. PMID- 3046451 TI - The corporate compromise: a Marxist view of health maintenance organizations and prospective payment. AB - Recent developments in health care are strikingly congruent with a Marxist paradigm. For many years small scale owner producers (physicians) dominated medicine, and the corporate class supported the expansion of services. As health care expanded, corporate involvement in the direct provision of services emerged. This involvement is reflected not only in the rise of for-profit providers, but also in the influence of hospital administrators, utilization review organizations, insurance bureaucrats, and other functionaries unfamiliar with the clinical encounter, but well versed on the bottom line. Corporate providers' quest for increasing revenues has brought them into conflict with corporate purchasers of care, whose employee benefit costs have skyrocketed. This intercorporate conflict powerfully shapes health policy and has caused the rapid proliferation of health maintenance organizations and other forms of prospective payment. Corporate purchasers of care favor the incentives under prospective payment for providers to curtail care and its costs. For corporate providers, prospective payment has allowed increased profits even in the face of constrained revenues, because reimbursement is disconnected from resource use. Unfortunately, this corporate compromise serves patients and physicians poorly. Alternative policy options that challenge corporate interests could save money while improving care. PMID- 3046453 TI - [Medico-Psychological Society. List of members]. PMID- 3046454 TI - [Chronic delusions and personality development. Historical outline]. PMID- 3046455 TI - Byssinosis in Belfast ropeworks: an historical note. PMID- 3046452 TI - [Contributions and difficulties of the social approach in psychiatry: apropos of the American Forces and veterans in Vietnam. 3. Pathogenic and therapeutic problems in veterans]. AB - Especially among a wide psychiatric population, the risk of a mingling between etiology, pathogenesis and description, peculiarly between correlations and explanations, is an important stumbling-block. However, like as acute war diseases, the part of a previous psychical vulnerability appears to be less important than the classical one which is played by "stressors", chiefly in case of their summation. Here, interfere not only combat "traumas", but also the circumstances of the readjustment after returning. In spite of interesting contributions to the mechanisms of traumatic neurosis followed up after Vietnam conflict, the reflection about intrusive-repetitive syndrome does not seem to have given rise to new developments. Concerning the therapeutics, the opinions about the results achieved--particularly with psychotherapies, in fact delicate and protracted--are more pessimistic than that of the authors. PMID- 3046456 TI - [Treatment of the acute graft versus host skin reaction with cyclosporin and PUVA]. AB - Acute graft-versus-host reaction is usually managed by immunosuppressive therapy, and cyclosporin is one drug of choice. In some patients, cyclosporin alone cannot always control the evolution. The use of other immunosuppressive drugs could be considered but their toxicity and side-effects are such that a vital risk is introduced for the patient. Conversely, when the disease primarily affects the skin, photochemotherapy (PUVA) associated with cyclosporin appears to be effective and safe. PMID- 3046457 TI - [Bullous scleroderma with the histological appearance of lichen sclerosus et atrophicus]. PMID- 3046458 TI - [Dysphagia and weight loss disclosing cicatricial pemphigus]. PMID- 3046459 TI - [A case of orificial plasmacytosis with periodontal localization]. PMID- 3046460 TI - [Myositis ossificans circumscripta]. PMID- 3046461 TI - [Localized lipo-atrophies]. PMID- 3046462 TI - [Observations on a series of 118 cases of acute intestinal invagination]. PMID- 3046463 TI - [Role of chemotherapy in the treatment of esthesioneuroblastoma in children. Apropos of 3 case reports]. PMID- 3046464 TI - The feasibility of screening for abdominal aortic aneurysms in a district general hospital. AB - The feasibility of a screening programme for abdominal aortic aneurysms within a district general hospital population is explored, based on our current accepted knowledge of the natural history of this disease process. It is shown that ultrasound screening of males aged between 65 and 74 years, with elective repair of the aneurysms discovered, could save up to 20 lives per year in this district at a reasonable and justifiable cost. Moreover, such a programme would not place an unacceptable burden on existing radiological and surgical facilities. PMID- 3046466 TI - Loss of split thickness skin grafts due to non-group A beta-haemolytic streptococci. AB - Over a 17-month period 77 patients requiring a split skin graft for a burn injury have suffered loss of previously well taken graft due to the growth of a beta haemolytic streptococcus. Of these only 42 were streptococci of Lancefield group A (Streptococcus pyogenes); 16 were group B, 3 group C and 16 group G. Some strains of groups B, C and G produce cytopathic and spreading factors capable of destroying the new skin graft and regenerating epithelium. We suggest that the non-group A streptococci may be more pathogenic than previously recognised in this particular respect. PMID- 3046465 TI - Nalbuphine and pentazocine in an opioid-benzodiazepine sedative technique: a double-blind comparison. AB - Sedation by a combination of an opioid drug such as pentazocine with a benzodiazepine is commonly used for minor surgical and investigative procedures. Nalbuphine is a newer drug which, like pentazocine, is an opioid agonist antagonist. Its actions are similar, but it has theoretical advantages in its profile of cardiovascular side effects. Nalbuphine or pentazocine in combination with diazepam were compared as components of a sedative technique for invasive radiology. The doses used were in the ratio of 2.5:1--ie nalbuphine 0.2 mg kg-1 and pentazocine 0.5 mg kg-1. Both regimens gave satisfactory results, and no difference could be detected between them in terms of sedation, analgesic efficacy, cardiovascular or respiratory changes, or recovery. Nalbuphine provides a safe and effective alternative to pentazocine in this situation. The study confirmed the need for caution because of the respiratory depressant effects of both drugs. PMID- 3046467 TI - Bacterial colonisation of leg ulcers and its effect on the success rate of skin grafting. PMID- 3046468 TI - [Facial eczema in ruminants and sporidesmins]. PMID- 3046470 TI - Report and recommendations of the San Antonio conference on diabetic neuropathy. PMID- 3046469 TI - Gerstmann-Straussler-Scheinker disease: immunohistological and experimental studies. AB - The older brother of the patient from whom the Fukuoka-1 strain was isolated was found to have numerous kuru plaques, the main finding common to both siblings. Other clinicopathological features including spongiform change were absent in the older brother. Immunostaining using anti-kuru plaque core protein and anti-beta protein peptide revealed many kuru plaques and a few senile plaques in the older brother. Experimental transmission of the disease to laboratory animals was successful, using tissues from both siblings, through inoculation of fresh brain homogenates, purified prion protein, and formalin-fixed brain homogenates. Prion protein fractions from the patient's brain shortened the incubation periods and formalin-fixed mouse brains did not lengthen the periods. The disease in the two brothers can be classified as Gerstmann-Straussler-Scheinker disease, a familial variant of Creutzfeldt-Jakob disease. Gerstmann-Straussler-Scheinker disease manifests a variety of clinicopathological features. Immunohistological verification of kuru plaques has major diagnostic value in assessing dementia. PMID- 3046472 TI - Principles and practice of intraperitoneal therapy. PMID- 3046471 TI - Hepatic arterial infusion of chemotherapy for metastatic colorectal cancer in the liver: why, how and what? PMID- 3046473 TI - Intraperitoneal chemotherapy for gastrointestinal malignancies. PMID- 3046474 TI - Intra-arterial chemotherapy using a pump. PMID- 3046475 TI - Intraarterial chemotherapy without pump. PMID- 3046477 TI - Treatment of soft tissue malignancies of the extremities by regional perfusion. PMID- 3046476 TI - Intraportal chemotherapy for colorectal hepatic metastases. PMID- 3046478 TI - Limitations of regional chemotherapy: a short comment. PMID- 3046481 TI - Randomized clinical trial of rifampin-trimethoprim and sulfamethoxazole trimethoprim in the treatment of localized urinary tract infections. AB - To investigate whether 10 days of rifampin-trimethoprim (RIF-TMP) or 6 weeks of sulfamethoxazole-trimethoprim (SMX-TMP) would decrease the relapse rate in patients with acute uncomplicated upper urinary tract infections in comparison with 10 days of SMX-TMP, we randomized 189 patients to receive RIF-TMP or SMX-TMP in a ratio of 1:2. After the site of infection was established by the antibody coated bacterium (ACB) test, patients with upper-tract infections who received SMX-TMP were again randomized and received either a total of 6 weeks or 10 days of therapy. All patients who received RIF-TMP were treated for 10 days. Clinical and microbiological evaluations were repeated at 2 and 6 weeks posttreatment. Eighty-five patients (54 ACB positive) received 10 days of RIF-TMP, 71 patients (45 ACB positive) received 10 days of SMX-TMP, and 18 patients (18 ACB positive) received 6 weeks of SMX-TMP. The overall recurrence rates in patients who received 10 days of therapy were 32% for RIF-TMP and 23% for SMX-TMP (P = 0.13). There were 12 (14%) relapses in the RIF-TMP group compared with 2 (3%) relapses in the SMX-TMP group (P = 0.01). In patients with upper-tract infections, the relapse rates were not statistically significantly different (P = 0.13). There were two (11%) recurrences (one relapse and one reinfection) in the 6-week treatment group. This 6% relapse rate was not different from the 4% relapse rate observed in patients with upper-tract infections who received 10 days of SMX-TMP. The number of patients who discontinued treatment because of an adverse effect in the 6-week SMX-TMP treatment group was significantly greater than those in the 10 day SMX-TMP treatment group (P=0.003) and the RIF-TMP treatment group (P=0.05). Ten days of SMX-TMP was as effective as 6 weeks of SMP-TMP or 10 days of RIF-TMP in the treatment of uncomplicated upper urinary tract infections and caused the fewest untoward effects. PMID- 3046480 TI - Antimalarial agents: mechanism of chloroquine resistance. PMID- 3046479 TI - Antimalarial agents: mechanisms of action. PMID- 3046482 TI - Cloning and expression of the imipenem-hydrolyzing beta-lactamase operon from Pseudomonas maltophilia in Escherichia coli. AB - The L-1 penicillinase structural gene, blaS, from Pseudomonas maltophilia has been cloned into the vector pACYC184. The pMON01 recombinant plasmid selected by ampicillin resistance carried a 2.6-kilobase Sau3A fragment of P. maltophilia DNA and was confirmed to express L-1 beta-lactamase by comparative isoelectric focusing. A detailed physical map was constructed, and the blaS structural gene was localized with a 17-mer oligonucleotide mixed probe encoding the L-1 N terminal amino acid sequence. Induction studies confirmed constitutive expression. Isolation of a complete beta-lactamase operon was attempted by construction of a P. maltophilia genomic library into phage lambda 2001. A recombinant phage was selected by DNA hybridization, and the 13.4-kilobase DNA insert was physically mapped and subcloned into plasmid vectors. Expression and L 1 beta-lactamase synthesis were studied in Escherichia coli. PMID- 3046483 TI - Isolation and characterization of a penicillinase from Pseudomonas cepacia 249. AB - Pseudomonas cepacia has an inducible beta-lactamase which is responsible for its novel ability to catabolize beta-lactam compounds. The gene encoding this enzyme, penA, was cloned from a genomic library of P. cepacia 249 on the broad-host-range cosmid pLAFR. This separated the penA gene from the gene encoding a second beta lactamase in P. cepacia 249. Expression of penA was inducible in an Escherichia coli host strain by low levels of penicillin. The 33,500-molecular-weight enzyme had penicillinase activity not inhibited by clavulanic acid or sulbactam and was highly active against piperacillin and azlocillin. In comparison with other inducible beta-lactamases produced by gram-negative organisms, the penA enzyme had many properties which were similar to those of the penicillinase produced by Alcaligenes faecalis. It was unlike the ampC-type cephalosporinase produced by Pseudomonas aeruginosa. PMID- 3046484 TI - Comparative study of cephalexin hydrochloride and cephalexin monohydrate in the treatment of skin and soft tissue infections. AB - In two prospective, randomized multicenter double-blind studies with a dosage of either 250 mg given four times a day (study A) or 500 mg given two times a day (study B), the comparative efficacy and safety of cephalexin hydrochloride (LY061188; Keftab) and cephalexin monohydrate (Keflex) for treatment of skin and soft tissue infections were determined. In study A, 97 patients received cephalexin hydrochloride and 101 patients received cephalexin monohydrate. In study B, 75 patients received cephalexin hydrochloride and 70 patients received cephalexin monohydrate. Diagnoses included abscesses, cellulitis, wound infections, and infected dermatitis, and were comparable in the different treatment groups. Pathogens were isolated from 82% of patients enrolled; the majority of isolates were of Staphylococcus aureus, Streptococcus pyogenes, other staphylococcal species, and a few gram-negative bacteria. In study A, 68 of 71 (95.7%) evaluable patients who received cephalexin hydrochloride responded satisfactorily; 73 of 81 (90%) patients who received cephalexin monohydrate also responded satisfactorily. In study B, 56 of 58 (96.5%) evaluable patients who received cephalexin hydrochloride responded satisfactorily; 47 of 50 (94%) patients who received cephalexin monohydrate also responded satisfactorily. An adverse clinical event leading to discontinuation of the treatment drug developed in 17 of 343 (4.95%) patients in both studies. No differences were noted between the two drugs. Skin eruptions, pruritus, and mild gastrointestinal symptoms were the common adverse effects. These data suggest that cephalexin hydrochloride, a new formulation of cephalexin, is a safe and effective antimicrobial agent for treatment of a variety of skin and subcutaneous infections in a dosage of either 250 mg four times a day or 500 mg twice a day. PMID- 3046485 TI - Dispersal in Campylobacter spp. of aphA-3, a kanamycin resistance determinant from gram-positive cocci. AB - DNA annealing studies indicated that kanamycin resistance in Campylobacter strains from various geographical areas is encoded by a gene structurally related to aphA-3 of gram-positive cocci. This finding confirms the transfer of genetic material between gram-positive and gram-negative bacteria under natural conditions. PMID- 3046486 TI - Mutants of Escherichia coli defective in acid fermentation. AB - Wild type E. coli ferments glucose to a mixture of ethanol and acetic, lactic, formic, and succinic acids. Mutants defective in acid production have now been isolated, including those defective in lactate dehydrogenase (LDH) or with excess alcohol dehydrogenase. These mutations had no phenotype without a pfl mutation. Novel mutants affecting acetate metabolism were isolated by insertion of the fusion vector Mudl. These aceG mutants cannot grow anaerobically on glucose or aerobically on acetate yet lack the pleiotropic growth defects of previously known pta/ack mutants. In some genetic backgrounds acetate negative mutations suppress the growth defects of adh mutations. These results are discussed in terms of redox balance. PMID- 3046487 TI - Continuous-sterilization system that uses photosemiconductor powders. AB - We report a novel photochemical sterilization system in which Escherichia coli cells were sterilized with photosemiconductor powders (titanium oxide). For sterilization that could be used in practice, it was necessary to separate the TiO2 powders from the cell suspension. Therefore, semiconductor powders were immobilized on acetylcellulose membranes. We constructed a continuous sterilization system consisting of a TiO2-immobilized acetylcellulose membrane reactor, a mercury lamp, and a masterflex pump. As a result, under the various sterilization conditions examined, E. coli (10(2) cells per ml) was sterilized to less than 1% survival when the cell suspension flowed in this system at a mean residence time of 16.0 min under irradiation (1,800 microeinsteins/m2 per s). We found that this system was reusable. PMID- 3046488 TI - The mannoprotein of Saccharomyces cerevisiae is an effective bioemulsifier. AB - The mannoprotein which is a major component of the cell wall of Saccharomyces cerevisiae is an effective bioemulsifier. Mannoprotein emulsifier was extracted in a high yield from whole cells of fresh bakers' yeast by two methods, by autoclaving in neutral citrate buffer and by digestion with Zymolase (Miles Laboratories; Toronto, Ontario, Canada), a beta-1,3-glucanase. Heat-extracted emulsifier was purified by ultrafiltration and contained approximately 44% carbohydrate (mannose) and 17% protein. Treatment of the emulsifier with protease eliminated emulsification. Kerosene-in-water emulsions were stabilized over a broad range of conditions, from pH 2 to 11, with up to 5% sodium chloride or up to 50% ethanol in the aqueous phase. In the presence of a low concentration of various solutes, emulsions were stable to three cycles of freezing and thawing. An emulsifying agent was extracted from each species or strain of yeast tested, including 13 species of genera other than Saccharomyces. Spent yeast from the manufacture of beer and wine was demonstrated to be a possible source for the large-scale production of this bioemulsifier. PMID- 3046489 TI - Factors influencing Clostridium botulinum spore germination, outgrowth, and toxin formation in acidified media. AB - Clostridium botulinum type A spores were inoculated at a level of 10(7) spores per ml into sterile beef media with protein concentrations of 1, 2, 3, 4, or 6% and acidified to pH values of 2.01 to 4.75 with hydrochloric acid or 4.19 to 4.60 with citric acid. All experimental manipulations, including blending, acidification, inoculation, incubation (30 degrees C), and analyses, were conducted in an anaerobic chamber-incubator in which atmospheric oxygen levels were maintained below 2 ppm (2 microliters/liter). Under these strict anaerobic conditions (oxidation-reduction values in media ranging from -370 to -391 mV), C. botulinum spores were consistently found to germinate, grow, and produce toxin below pH 4.6. The boundary between toxic and atoxic samples in HC1-acidified beef media was mediated by titratable acidity, pH, and protein concentration. A limiting acidity was not established for the citrate-acidified samples; all blends tested (1, 2, 3, and 4% protein and titratable acidities of 0.091 to 0.453%) became toxic within 5 weeks. At the same pH and protein concentration, citric acid was less effective than HC1 in preventing the germination of C. botulinum spores. Higher levels of cell proliferation in the beef protein, as well as enhanced gas production and putrefactive degradation, indicated that beef was a better substrate than soy for C. botulinum spores under these conditions. Reducing the inoculum to 10(4) delayed but did not prevent spore outgrowth and toxin release at pH levels below 4.6. PMID- 3046490 TI - National field evaluation of a defined substrate method for the simultaneous enumeration of total coliforms and Escherichia coli from drinking water: comparison with the standard multiple tube fermentation method. AB - A defined substrate method was developed to simultaneously enumerate total coliforms and Escherichia coli from drinking waters without the need for confirmatory or completed tests. It is a new method based on technology that uses a hydrolyzable substrate as a specific indicator-nutrient for the target microbes. No equipment other than a 35 degrees C incubator and long-wavelength (366-nm) light is necessary. To perform the test, one only has to add water to the powdered ingredients in a tube or flask. If total coliforms are present in the water sample, the solution will change from its normal colorless state (no target microbes present) to yellow. The specific presence of E. coli will cause the same tube to fluoresce under a longwave (366-nm) UV lamp. The test, called Autoanalysis Colilert (AC), was compared with Standard Methods for the Examination of Water and Wastewater 10-tube multiple tube fermentation (MTF) in a national evaluation. Five utilities, representing six U.S. Environmental Protection Agency regions, participated. All water samples came from distribution systems. Split samples from a wide variety of water sources were analyzed for the MPN-versus-MPN comparison. A total of 1,086 tubes were positive by MTF, and 1,279 were positive by AC. There was no statistical difference between MTF and AC. Species identifications from positive tubes confirmed the sensitivity of the AC. A national evaluation of the AC test showed that it: (i) was as sensitive as Standard Methods MTF, (ii) specifically enumerated 1 total coliform per 100 ml, in a maximum of 24 h, (iii) simultaneously enumerated 1 E. coli per 100 ml in the same analysis, (iv) was not subject to false-positive or false-negative results by heterotrophic bacteria, (v) did not require confirmatory tests, (vi) grew injured coliforms, (vii) was easy to inoculate, and (viii) was very easy to interpret. PMID- 3046491 TI - Transduction of Escherichia coli by bacteriophage P1 in soil. AB - Transduction of Escherichia coli W3110(R702) and J53(RP4) (10(4) to 10(5) CFU/g of soil) by lysates of temperature-sensitive specialized transducing derivatives of bacteriophage P1 (10(4) to 10(5) PFU/g of soil) (P1 Cm cts, containing the resistance gene for chloramphenicol, or P1 Cm cts::Tn501, containing the resistance genes for chloramphenicol and mercury [Hg]) occurred in soil amended with montmorillonite or kaolinite and adjusted to a -33-kPa water tension. In nonsterile soil, survival of introduced E. coli and the numbers of E. coli transductants resistant to chloramphenicol or Hg were independent of the clay amendment. The numbers of added E. coli increased more when bacteria were added in Luria broth amended with Ca and Mg (LCB) than when they were added in saline, and E. coli transductants were approximately 1 order of magnitude higher in LCB; however, the same proportion of E. coli was transduced with both types of inoculum. In sterile soil, total and transduced E. coli and P1 increased by 3 to 4 logs, which was followed by a plateau when they were inoculated in LCB and a gradual decrease when they were inoculated in saline. Transduction appeared to occur primarily in the first few days after addition of P1 to soil. The transfer of Hg or chloramphenicol resistance from lysogenic to nonlysogenic E. coli by phage P1 occurred in both sterile and nonsterile soils. On the basis of heat induced lysis and phenotype, as well as hybridization with a DNA probe in some studies, the transductants appeared to be the E. coli that was added. Transduction of indigenous soil bacteria was not unequivocally demonstrated.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3046492 TI - Mechanism for nitrosation of 2,3-diaminonaphthalene by Escherichia coli: enzymatic production of NO followed by O2-dependent chemical nitrosation. AB - The mechanism by which Escherichia coli can catalyze the nitrite-dependent nitrosation of 2,3-diaminonaphthalene (DAN), with formation of the corresponding fluorescent triazole, was studied. The reaction was dependent on production of a gaseous compound which can nitrosylate DAN upon contact with air. This compound was identified as nitric oxide (NO), and the kinetics of NO and triazole production are reported. NO and triazole were produced proportionally in a stoichiometric ratio, NO/triazole, of 1.4 to 1.7. Given the requirement for air, nitrosation of DAN probably proceeds via formation of the well-known strong nitrosylating agents N2O3 and N2O4 from NO. The parallel inhibition of NO and triazole production by azide and nitrate served to reinforce the link between nitrosation and nitrate reductase that had been established previously by others on genetic grounds. PMID- 3046493 TI - Phenotypic and genotypic characterization of tetracycline- and oxytetracycline resistant Aeromonas hydrophila from cultured channel catfish (Ictalurus punctatus) and their environments. AB - Oxytetracycline-resistant (OTcr) and tetracycline-resistant (Tcr) Aeromonas hydrophila were isolated commonly from catfish intestinal contents and the water and sediment of catfish culture ponds, but less frequently in market catfish. Isolates demonstrated two resistance phenotypes, Tcr OTcr and Tcs OTcr, when plated directly on to MacConkey agar containing 30 micrograms of tetracycline or oxytetracycline per ml. Tcs OTcr isolates expressed Tcr after induction by 1 h of growth in tryptic soy broth containing 1 microgram of tetracycline per ml. Over 90% of the resistant aeromonads hybridized with DNA probes for class A or class E Tcr determinants; class E was twice as prevalent as class A. The distribution of class A and E Tcr determinants varied with the Tcr phenotypes. Prior to induction, 86% of isolates with class A determinants were Tcr as well as OTcr, while only 16% with class E determinant were Tcr. Three of 5 Tcr aeromonads without class A or class E determinants and 1 of 14 with class E determinants transferred Tcr to Escherichia coli. PMID- 3046494 TI - Novel compound for identifying Escherichia coli. AB - A new chromogenic compound, 5-bromo-4-chloro-3-indoxyl-beta-D-glucuronide, was found to be useful for the rapid, specific, differential identification of Escherichia coli in the sanitary analysis of shellfish and wastewater. Of 1,025 presumptively positive colonies (blue) and 583 presumptively negative colonies (nonblue), only 1% false-negative and 5% false-positive results were found. PMID- 3046495 TI - [A case of non-Hodgkin lymphoma with complete remission achieved by carboplatin]. AB - The patient was a 63-year-old female, who was admitted to the National Nagoya Hospital with complaints of left cervical and bilateral axillary lymphadenopathy on 12 May, 1987. Since May 1985, with the diagnosis of non-Hodgkin lymphoma, she had been successfully treated with combination chemotherapy (VEPA) and radiotherapy at another hospital. Left axillary lymphode biopsy revealed a diagnosis of non-Hodgkin lymphoma, diffuse large cell type. Then, she was given intravenous administration of carboplatin (400 mg/body) on 26 May, 1987. After a single course of this regimen, the lymphnode swelling subsided, and she achieved complete remission on 6 June. Thereafter, she was placed on the maintenance chemotherapy of carboplatin (400 mg/body) every 5 weeks. Through the whole course of this patient, the serum levels of blood urea nitrogen and creatinine were normal, and she did not notice nausea, vomiting and peripheral neuropathy. To our knowledge, this is the first report of complete response by the administration of carboplatin for non-Hodgkin lymphoma. PMID- 3046497 TI - The cutaneous immunopathology of necrobiosis lipoidica diabeticorum. AB - Twelve female patients with necrobiosis lipoidica diabeticorum (six with diabetes and six without) had a 5-mm punch biopsy of the skin lesion performed. The tissue was processed for dermatopathologic examination in 12 cases and for direct immunofluorescence in 11. Vasculopathy with inflammation and thickening of vessel walls, at times leading to occlusion, was found in lesional skin in all 12 cases. Vessels contained deposits of immunoreactants in the involved skin in 11 cases. This included IgM in six, C3 in nine, fibrin in ten, IgG in one, and IgA in two. Vessels contained deposits of immunoreactants in uninvolved skin in seven patients (C3 in four, IgM in three, fibrin in three, C4 in one, and IgA in one), three of whom had type I diabetes. PMID- 3046496 TI - [Pirarubicin (THP-adriamycin)]. AB - Pirarubicin (THP-adriamycin or THP-doxorubicin) was found during a search of new anthracycline antibiotics among 4'-O-substituted compounds having less toxicities than other anthracycline anticancer drugs in 1979 by Umezawa et al. In its preclinical studies, this compound possessed almost similar antitumor efficacies to doxorubicin, but was effective against doxorubicin-resistant P388 and other murine tumor cell lines. This compound was rapidly incorporated into tumor cells, inhibiting DNA polymerase alpha and subsequently DNA synthesis. Inhibition of RNA synthesis was also noted. In the clinical studies, clinical responses were established against head and neck cancer, breast cancer, urogenital cancers, ovarian cancer, uterine cancer, acute leukemia, and malignant lymphoma, showing a wide antitumor spectrum clinically. Among the side effects, cardiac toxicity, alopecia and disturbance of the digestive organs were mild. From these results, THP-adriamycin seems to be a useful clinical drug for human solid tumors. PMID- 3046498 TI - Conidiobolus coronatus infection treated with ketoconazole. AB - We describe a case of subcutaneous zygomycosis caused by Conidiobolus coronatus of six years' duration. Following treatment failure using potassium iodide, the patient responded with ketoconazole therapy. She remained well three years following therapy. Brazilian cases of subcutaneous infections caused by C coronatus and Basidiobolus ranarum are reviewed. PMID- 3046499 TI - Mucous gland basement membrane immunofluorescence in cicatricial pemphigoid. AB - Three patients are described with cicatricial pemphigoid and positive immunofluorescence findings in the basement membrane zone of mucous glands of the pharynx, mouth, and nose. These findings appear to be unique to cicatricial pemphigoid. PMID- 3046500 TI - Immunofluorescence, necrobiosis lipoidica, and blood vessels. Fluorescent lights in the tunnels. PMID- 3046501 TI - Acute follicular graft-vs-host disease. PMID- 3046502 TI - Papular or papulovesicular syndromes. PMID- 3046503 TI - Inherited peroxisomal disorders involving the nervous system. PMID- 3046504 TI - Clinical significance of IgG subclass deficiency. PMID- 3046505 TI - Antineutrophil cytoplasm antibody in crescentic glomerulonephritis. AB - Antineutrophil cytoplasm antibody (ANCA) has been reported in the sera of adults with Wegener's granulomatosis and microscopic polyarteritis, but this phenomenon has not so far been described in children. We report three children with crescentic glomerulonephritis in whom ANCA concentrations were raised at presentation. Two cases were idiopathic, and the other later developed features of Wegener's granulomatosis. In all three, plasma exchange and immunosuppression removed ANCA, but in only one case was there clinical improvement. This child later developed classical nasal lesions of Wegener's granulomatosis associated with a decline in renal function and a rise in ANCA. Plasma exchange and immunosuppression again produced a good clinical response and removed ANCA. This suggests that ANCA is either a marker of disease activity, or is involved in the pathogenesis of disease. PMID- 3046506 TI - Prenatal diagnosis of cystic fibrosis. PMID- 3046507 TI - Dilemmas associated with antenatally detected urinary tract abnormalities. AB - Over a five year period 55 fetuses had abnormalities of the urinary tract detected by antenatal ultrasound scan. The incidence was 1:935 total births during a one year prospective study. Intrauterine intervention was undertaken in five for suspected obstructive uropathy, which was confirmed in only two. Of 51 live born infants, five died (two with renal failure), and only 18 (35%) had a clinically detectable abnormality at birth. Twenty seven patients underwent postnatal operations, the remainder being treated conservatively. Antenatal counseling was seldom undertaken by those responsible for the postnatal care. There were many instances of prospective parents receiving little or inappropriate information. Greater cooperation is required between all the staff concerned particularly as the natural history and appropriate postnatal management of some urinary tract abnormalities are still not known. PMID- 3046508 TI - Neonatal ovarian cysts: therapeutic dilemma. AB - Seven cases of neonatal ovarian cysts that presented over the past seven years were studied. Complications included torsion and rupture and usually occurred in cysts more than 5 cm in diameter. Surgical removal, either oophorectomy or cystectomy, was the treatment of choice. Because even cystectomy results in loss of normal ovarian tissue, and because spontaneous regression of cysts less than 5 cm in diameter can occur, a more conservative approach is now proposed. Regular ultrasonography alone is recommended if the cysts are less than 5 cm in diameter, and aspiration of the cysts followed by regular ultrasonographs if the cysts are more than 5 cm in diameter. Operation should be reserved for recurrent cysts or for those with complications. Cysts diagnosed antenatally may be aspirated in utero if there are signs of thoracic compression. PMID- 3046509 TI - Prenatal diagnosis and carrier detection in primary immunodeficiency disorders. PMID- 3046510 TI - Bone marrow transplantation for leukaemia. PMID- 3046511 TI - Mitochondrial DNA and genetic disease. PMID- 3046512 TI - Phototherapy: an ocular hazard revisited. PMID- 3046514 TI - IgG antibodies to Aspergillus fumigatus in cystic fibrosis: a laboratory correlate of disease activity. AB - Serum was collected from 50 patients with cystic fibrosis, and IgG antibodies to Aspergillus fumigatus were measured by enzyme linked immunosorbent assay (ELISA). In addition, total IgE and Aspergillus specific IgE antibodies were measured in 41 of the 50. A close association was found between pulmonary function and clinical state, and IgG antibodies to Aspergillus. There was no association between pulmonary function or clinical state and IgE antibodies. It is postulated that in patients with cystic fibrosis, Aspergillus fumigatus may contribute to deterioration in pulmonary function by local pathogenicity, or by hypersensitivity mechanisms mediated by IgG. PMID- 3046513 TI - Air or oxygen as driving gas for nebulised salbutamol. AB - The effects of nebulised salbutamol driven by compressed air or oxygen were compared in a randomised crossover study during 27 attacks of acute asthma. Arterial oxygen saturation fell by 2-6% during or after treatment in 10 cases: seven with compressed air, two with oxygen, and one with both driving gases. Hypoxaemia occurred in younger children and in those who fell asleep, but was not related to the level of arterial oxygen saturation before treatment or the size of the response to bronchodilator therapy. More children fell asleep with compressed air nebulisation. Arterial oxygen saturation improved and heart rates remained stable during treatment when oxygen was the driving gas. After treatment, however, arterial oxygen saturation fell and heart rates rose to values that were similar to those after treatment with compressed air. The falls in arterial oxygen saturation we observed, though comparatively small, would be clinically important on the steep part of the oxygen dissociation curve, and our results emphasise that families with home nebulisers should seek medical advice early when their children develop severe asthma. The benefits of using oxygen as the driving gas during nebulisation were transient, and in severe asthma treatment with oxygen needs to be continued after the nebulised salbutamol has been given. PMID- 3046515 TI - Clinical outcome of fetal uropathy. AB - Thirty seven infants (27 boys and 10 girls) whose uropathy had been diagnosed antenatally were reviewed at a mean age of 24.4 months. Antenatal ultrasonography was found to be an accurate detector of renal disease, as uropathy was subsequently confirmed in 33 of the 37 infants (89%). A smaller proportion of patients required operations than in other series reported. Shortly after birth the kidney undergoes physiological adjustments and investigation may be carried out too soon. It should be delayed, especially if the infant is well. The timing of postnatal ultrasonography is important and the advice of a paediatric urologist should be sought before investigations are carried out. PMID- 3046516 TI - Ribavirin in respiratory syncytial virus infection. A double blind placebo controlled trial is needed. PMID- 3046517 TI - Renal failure in the newly born. PMID- 3046518 TI - Is the gut intrinsically abnormal in rheumatoid arthritis? PMID- 3046520 TI - Experimental repair of ventricular septal defects using autologous right ventricular muscle flaps: preliminary report. AB - Survival after repair of postinfarction ventricular septal defects remains poor, often due to extensive loss of contractile muscle in the septum or left ventricle. We evaluated whether a contractile flap of right ventricular muscle could be used to repair a similar ventricular septal defect to augment left ventricular performance in 7 fully instrumented mongrel dogs (weight, 23 to 28 kg). By using hypothermic bypass and cold fibrillatory arrest, a trapezoidal right ventricle flap was fashioned from the free wall of the mid to lower right ventricle, basing its widest portion anteriorly on the septum and left ventricle. A large, 2-cm-diameter core of septum was excised beneath this flap to simulate a postinfarct ventricular septal defect. The right ventricular flap was then invaginated through the defect and sewn to the left ventricular side of the septum with pledgeted sutures taken full thickness through the flap and septum in a "vest-over-pants" fashion. Contraction of the right ventricular flap was confirmed visually and by postbypass multiple gated acquisition scans. The right ventricular defect was closed with fascia lata. All dogs were weaned from bypass without inotropes. Precardiac and postcardiac outputs of 2.5 +/- 0.5 versus 2.3 +/- 0.4 L/min and left ventricular end-diastolic pressures of 4 +/- 2 versus 4 +/ 3 mm Hg were identical. No shunts were detected by oxygen saturation. Autopsies confirmed the integrity of the repair. We conclude that septal defects can be repaired by using contractile right ventricular muscle, thus preserving left ventricular function. This technique offers promise for repair of postinfarction ventricular septal defects by using autologous, already conditioned to contract, cardiac muscle, but its application in humans must await long-term testing. PMID- 3046519 TI - Rheumatoid arthritis and gut related lymphocytes: the iteropathy concept. PMID- 3046521 TI - Total obstruction of the left main coronary artery. AB - A rare and often fatal condition, total obstruction of the left main coronary artery has been treated with increasing success in both acute and chronic clinical settings. Seventeen patients with acute occlusion have been reported in the literature. All were discovered at the time of acute periinfarction catheterization and were treated aggressively with intracoronary thrombolysis, percutaneous transluminal angioplasty, emergency bypass surgery, or a combination of techniques. Chronic total occlusion of the left main coronary artery has been reported in 59 patients, including 3 at our institution. These patients present with chronic but increasingly severe angina. A right dominant coronary anatomy is always found, usually with well-developed right-to-left collaterals. The results of surgical revascularization in our 3 patients and in 45 others described in the literature support the safety and efficacy of this approach. PMID- 3046522 TI - Cardiac transplantation: the ideal myocardial temperature for graft transport. AB - The ideal preservation method and cooling temperature for transport of donor hearts are not known. Serious derangements in myocardial relaxation are well described with different methods of cooling. To assess this problem, human right atrial trabeculae contracting isometrically at 34 degrees C in vitro were subjected to hypothermic arrest at 1, 4, 12, and 20 degrees C for 1, 2, 4, 24, and 48 hours. Control conditions were resumed, and myocardial mechanical recovery was assessed over 1 hour. Contraction was 50% depressed after a 1- to 2-hour exposure to 1 degree C and was almost completely arrested following a 4-hour exposure. Muscles cooled to 4 degrees C recovered poorly, whereas those cooled to 12 and 20 degrees C did well. In the latter 2 groups, force development increased rapidly on rewarming and exceeded the precooling contraction force (p less than 0.05). A 100% increase in relative resting force was seen in muscles cooled to 1 and 4 degrees C (p less than 0.05). This finding suggests a failure of calcium homeostasis at very low temperatures. We conclude that atrial preservation is optimal at about 12 degrees C. PMID- 3046523 TI - Thoracic hydatid cysts: a report of 842 cases treated over a thirty-year period. AB - From 1957 to 1985, 842 patients were diagnosed as having thoracic hydatid cysts; 810 cysts were intrathoracic, 29 occurred on the "liver roof," 2 were cardiac, and 1 was on the chest wall. A total of 1,010 surgical procedures were performed in 807 patients (35 refused operation). There was a total operative mortality of 0.6% (5 deaths). Procedures became more conservative as experience was gained, and 79% of the procedures were endocystectomies. Intact endocystectomy (Barrett's technique) without preliminary aspiration was the approach of choice. Careful protection of the operating field, suturing of all the bronchial openings, and capitonnage were the keys to successful treatment. One hundred six patients with intact endocystectomies done before July, 1975, were followed for 3 to 20 years. Ruptures occurred during cyst manipulation in 35 patients (33%). Recurrence after operation was seen in 2 patients (1.9%). There were no deaths among the patients undergoing intact endocystectomy. In comparison, we followed 136 patients who underwent aspiration endocystectomy and the recurrence rate was 3.7% (5 patients). PMID- 3046524 TI - Acute purulent mediastinitis and sternal osteomyelitis after closed chest cardiopulmonary resuscitation: a case report and review of the literature. AB - Numerous complications have been associated with cardiopulmonary resuscitation. Acute purulent staphylococcal mediastinitis and sternal osteomyelitis are, however, unusual and do not appear to have been reported previously in association with closed chest resuscitation. Sternal fracture during chest compressions and subsequent hematogenous seeding of the resultant retrosternal hematoma with Staphylococcus aureus led to purulent mediastinitis and sternal osteomyelitis in our patient. The source of bacteremia may have been a resolving phlebitis at an intravenous catheter insertion site. Early diagnosis, aggressive surgical debridement, and antibiotic therapy were key to a successful outcome. PMID- 3046526 TI - A single pericardial patch technique for repair of partial anomalous pulmonary venous drainage associated with sinus venosus atrial septal defect. AB - A technique is described for repair of partial anomalous pulmonary venous drainage associated with sinus venosus atrial septal defect. The procedure, using a single autologous pericardial patch, is able to facilitate both reconstruction of the pulmonary venous channel and enlargement of the superior vena cava. This technique also reduces the incidence of arrhythmias. PMID- 3046528 TI - The isolated pulmonary nodule. PMID- 3046527 TI - Neoadjuvant therapy of stage III non-small cell lung cancer. AB - During the past several years, there has been a resurgence of interest in preoperative or postoperative chemotherapy in patients with Stage III lung cancer. The staging system for lung cancer has recently been modified, and at the present time Stage III disease is now subdivided into Stage III-A (potentially surgically resectable for cure) and Stage III-B (unresectable). This article will review five recently completed studies utilizing neoadjuvant therapy in various types of Stage III lung cancer. The thoracic surgeon is faced with the dilemma of reviewing this literature and trying to make a conclusion as to what is appropriate therapy for Stage III disease. Unquestionably, neoadjuvant therapy appears to increase the resectability rate in Stage III-A disease and can make some Stage III-B patients anatomically resectable. It is hoped that future well designed phase III studies can be accomplished in Stages III-A and III-B disease so that we can determine whether neoadjuvant chemotherapy is or is not beneficial for these patients. PMID- 3046529 TI - Adjustable annuloplasty for tricuspid insufficiency. PMID- 3046525 TI - Separate extraction of cardiac and pulmonary grafts from a single organ donor. AB - The scarcity of multiple-organ donors for perfused organ transplantation requires cooperation between various transplanting teams to maximize organ retrieval. We have developed a technique for the extraction of a cardiac and separate pulmonary graft from the same donor. These grafts can then be successfully implanted into two separate recipients. Our experience with 9 successful extractions and implantations is recorded. PMID- 3046530 TI - Muscle sparing thoracotomy. PMID- 3046531 TI - [New antineoplastic antibiotics of the anthracycline group and the prospects of their clinical use]. PMID- 3046532 TI - [The use of electromagnetic fields of ultrahigh frequency range in biotechnology]. PMID- 3046533 TI - [Nucleosides in the chemotherapy of viral infections]. PMID- 3046535 TI - E-rosettes in aging: meta-analysis of the literature. AB - In this meta-analysis of the literature, it is affirmed that aging is associated with a small but significant decline in peripheral E-rosette-forming cells while high affinity E-rosettes (active or early rosettes) increase. The author speculates that this differential age effect may reflect the abnormal capping behaviour of E-rosettes or a change in the cell surface charge properties. A type II error is a common trait of the reviewed studies and could well explain the great number of reports concluding that E-rosette-forming cells are unaltered in the elderly. PMID- 3046534 TI - Systemic treatment of cancer in the elderly. AB - The goal of this review is to provide a readable and exhaustive reference in three major areas of geriatric oncology: complications of chemotherapy and radiotherapy, responsiveness of cancer to systemic treatment, social issues in the care of elderly patients with terminal illnesses. The conclusions of this study are: 1. Progressive deterioration of renal function is the most consistent change of aging. Adjustment of doses of renally excreted drugs to individual creatinine clearance may prevent life-threatening myelotoxicity in the elderly. 2. Intensive chemotherapy regimens (acute leukemia, non Hodgkin's lymphoma) cause more serious and prolonged myelotoxicity in the elderly. Elderly are more susceptible than younger patients to cardiotoxicity and central and peripheral neurotoxicity. Age is a poor predictor of complications in other organs or systems. 3. The prognosis of patients with Hodgkin's disease worsens with aging, possibly due to increased prevalence of mixed cellularity histology. It is controversial whether the prognosis of other neoplasias is poorer. Prognosis is not age-related in multiple myeloma. In general, elderly in good performance status may benefit from systemic cancer treatment to the same extent as younger patients, except for Hodgkin's disease. 4. The Informal Support Network, epitomized by the family, appears the most suitable environment to care for the elderly with cancer. PMID- 3046536 TI - The diagnosis of reversible dementia in the elderly. A critical review. AB - Evaluation of the elderly demented patient includes a search for treatable diseases; recommendations for this approach have been based on reports of a high prevalence of "reversible" dementia in several series of patients in the medical literature. A critical review of the methodology of these reports was undertaken to determine their validity as sources for the protocols often recommended. Several methodologic flaws were found in many studies that recommended extensive "screening" of the demented elderly, limiting their usefulness as resources. More carefully designed studies are needed to understand better the frequency and etiology of reversible causes of dementia. PMID- 3046537 TI - Acute response to bronchodilator. An imperfect guide for bronchodilator therapy in chronic airflow limitation. AB - We conducted a four-period cross-over randomized trial in which we found that patients with chronic airflow limitation demonstrated symptomatic improvement with both inhaled albuterol and oral theophylline. The response, however, was not uniform. We therefore tested the ability of acute change in forced expired volume in one second (FEV1) following inhaled beta agonist to predict long-term symptomatic response to albuterol and theophylline. We found that the reproducibility of acute change in FEV1 over three repetitions was poor (intraclass correlation 0.17). Furthermore, the mean improvement FEV1 following inhaled albuterol across the three repetitions did not relate closely to symptomatic response to either albuterol or theophylline. We conclude that acute response to inhaled beta agonist is not useful for identifying patients with chronic airflow limitation who are likely to benefit from bronchodilator treatment. PMID- 3046538 TI - Verapamil vs quinine in recumbent nocturnal leg cramps in the elderly. AB - In an open-labeled trial with eight elderly patients (aged 62 to 87 years) suffering from nocturnal leg cramps refractory to treatment with quinine sulfate, we ruled out other active disease processes and substituted verapamil hydrochloride therapy (120 mg at bedtime). Response to treatment was assessed by biweekly observations by the primary care physician and nightly by the research registered nurse for the entire duration of the trial, lasting eight weeks. Observations made and clinical conditions reported were indicative of improvement and disappearance of cramping when therapy was changed from quinine to verapamil. This noteworthy improvement in patients with recumbent nocturnal leg cramps is an important finding and merits further investigation. PMID- 3046539 TI - Leprosy in six isolated residents of northern Louisiana. Time-clustered cases in an essentially nonendemic area. AB - Northern Louisiana has been essentially free of indigenous leprosy, and now it is not. Six new cases of leprosy have been diagnosed: three in 1986, the other three in 1985, 1983, and 1982, respectively. The patients had been lifelong residents of six scattered rural parishes. Leprosy had never been reported from five of them. No patient had had contact with human leprosy. The patients were white; four were women; the mean +/- SD age at onset was 60.3 +/- 16.4 years (age range, 31 to 80 years); and the mean +/- SD interval to diagnosis was 1.2 +/- 1.4 years. One patient had Hodgkin's disease at the age of 25 years and leprosy at the age of 31 years; another patient had cervical carcinoma. All rural northern Louisiana residents coexist with armadillos (Dasypus novemcinctus), some of which are infected with Mycobacterium leprae, the significance of which is unknown. Hypothetically, exposure to an unknown human case, reactivation of "asymptomatic" leprosy through immunosenescence or immunosuppression, or infection from an environmental source might have occurred. Because the patients lacked contact, travel, residence, and exposure risk factors, the origin of leprosy in the new indigenous cases is noteworthy and is not understood. PMID- 3046540 TI - Labeling of participants in high blood pressure screening programs. Implications for blood cholesterol screenings. AB - Screening programs have expanded to identify the many persons who are unaware of their high blood cholesterol level and thus are at an increased risk for coronary heart disease. These programs bring both potential benefits and potential risks to the participant. One potential risk is that of iatrogenic effects of learning one's risk status, often referred to as the "labeling phenomenon." Research that has addressed the labeling phenomenon in blood pressure screening programs has important implications for blood cholesterol screenings. Detrimental effects on screening participants are possible, but they can be attenuated by careful attention to characteristics of the debriefing and counseling that should be included in screening protocols. PMID- 3046541 TI - Precipitating factors leading to decompensation of heart failure. Traits among urban blacks. AB - Potential precipitating factors that led to cardiac decompensation and subsequent hospital admission for heart failure were examined in 101 patients in a large public hospital serving a predominantly working-class minority population. Ninety seven percent of patients were black; their age was 59 +/- 14 years (mean +/- SD); on average, they were hospitalized three times in the preceding year for problems related to their heart failure. Potential precipitating factors for decompensated heart failure were identified in 93% of patients. Lack of adherence to the prescribed medical regimen was the most commonly identified causative factor and was noted in 64% of the cases; noncompliance with diet amounted to 22%, with drugs to 6%, and with the combination of drugs and diet to 37%. Other factors also related to hospitalization were cardiac arrhythmias (29%), emotional/environmental issues (26%), inadequately conceived drug therapy (17%), pulmonary infections (12%), and thyrotoxicosis (1%). Thus, the key preventive measure necessary in at least two thirds of patients centered around better adherence to drug and/or diet regimen, highlighting the precept that better patient education is mandatory if we are to minimize the number of hospital admissions for decompensated heart failure. PMID- 3046542 TI - Effects of transdermal clonidine treatment on withdrawal symptoms associated with smoking cessation. A randomized, controlled trial. AB - In a double-blind, randomized, placebo-controlled trial, we studied 40 cigarette smokers to determine the effects of one week of transdermal clonidine hydrochloride (Catapres-TTS No. 2) treatment on the withdrawal symptoms associated with smoking cessation. Subjects were instructed to maintain their usual cigarette intake during days 1 through 3 and cease smoking for days 4 through 6. All of the withdrawal symptoms measured (craving, irritability, anxiety, restlessness, difficulty concentrating, and hunger) significantly increased during the three days of smoking cessation in the placebo group. There was a 4.3-fold increase in craving, a 3.8-fold increase in irritability, a 3.7 fold increase in anxiety, and a 3.3-fold increase in restlessness in the placebo group compared with the transdermal clonidine group during the three days of smoking cessation. Impairment of concentration and hunger were not significantly diminished by transdermal clonidine treatment during smoking cessation. In addition, a trend was present in the transdermal clonidine group to spontaneously decrease the number of cigarettes smoked per day during the smoking period. Side effects were generally mild. We conclude that transdermal clonidine treatment ameliorates some of the short-term withdrawal symptoms, especially craving, associated with smoking cessation. PMID- 3046543 TI - Early metastatic cancer of unknown primary origin at presentation. A clinical study of 302 consecutive autopsied patients. AB - We studied 302 consecutive autopsied patients who presented with carcinoma of unknown primary origin. The most frequent metastatic sites were the nodes, lung, and bone. The primary site was identified while patients were alive in 27% and at autopsy in 57%; the site remained unidentified in 16%. The pancreas (26.5%), lung (17.2%), kidney (4.6%), and colorectum (3.6%) were the most frequent primary sites, but the reliability of diagnostic tests used in the search for this site was disappointing. Survival was identical in patients whose primary site was discovered while alive, at autopsy, or remained unknown. The number of metastases at presentation was the major prognostic factor. Analysis of autopsy data demonstrated that patients with carcinoma of unknown primary origin pursue a different course than expected when the primary site is the first manifestation of the disease. On the basis of these results and the results of other modern series, we suggest an approach consisting of a limited initial workup but with greater emphasis on modern histochemistry studies and immunohistopathologic and other kinetic and morphologic parameters to understand the patient tumor characteristics better and base the clinical management on an individual basis. PMID- 3046545 TI - Austin Flint--America's Laennec revisited. PMID- 3046544 TI - The biological actions and potential clinical significance of dietary omega-3 fatty acids. AB - The exploration of the effects of fish oil in cardiovascular disease and inflammation seems to be a promising avenue of research. There is evidence indicating the role of marine oils in inhibiting coagulation and platelet, leukocyte, and T-lymphocyte function. Moderate amounts of fish oil reduce serum triglyceride and very-low-density lipoprotein levels, while effects on cholesterol, high-density lipoprotein, and low-density lipoprotein levels are unpredictable except at extremely large doses. Putative actions in lowering blood pressure and limiting myocardial infarct size require further study. The clinician needs to be aware of the dose- and time-dependent nature of the measurable effects of fish oil. The folly of recommending two to four capsules per day is contrasted with the ten to 30 capsules required to produce a specific desired effect. PMID- 3046547 TI - Herpes simplex lymphangitis. Two cases and a review of the literature. AB - Lymphangitis and lymphedema are rarely reported complications of herpetic hand or genital infection. The natural history of these complications is gradual resolution over 14 to 21 days. Recognition of this presentation of herpes infection avoids unnecessary surgery and antibacterial therapy. Antiviral therapy may have a role in shortening the duration of symptoms and aborting recurrent lymphangitic episodes. PMID- 3046546 TI - Management of cancer during pregnancy. AB - Although cancer during pregnancy is infrequent, its management is difficult for patients, their families, and their physicians. When termination of the pregnancy is unacceptable, decisions regarding the use of irradiation and chemotherapy are complicated by the well-known high risks of abortion and fetal malformation. This risk is concentrated in the first trimester and varies with the choice of chemotherapeutic agents or combinations of agents. There is only minimal evidence of increased risk of malformation or abortion in the second or third trimester. Recent progress in cancer therapy has made cure a reasonable goal, and for some malignant neoplasms, cure is still possible even when initial therapy is modified or delayed. When cure is a reasonable goal, curative therapy should not be compromised by modification or delay. When treatment for cure or significant palliation is not possible, however, the goal should shift to protection of the fetus from damage by the injudicious use of teratogenic cancer therapy. This report will review the available data that may assist in these difficult decisions. PMID- 3046548 TI - Efficacy of computer systems in aiding in the clinical management of patients. PMID- 3046549 TI - [Recurrent infections caused by Salmonella typhimurium]. AB - We report the case of a 27 month-old girl with recurrent Salmonella typhi murium infections. The study of the neutrophil cells showed a transitory selective deficiency of the chemiluminescent response. After 5 months of pefloxacine therapy, recovery was obtained. PMID- 3046550 TI - [Esophageal duplication with pleural effusion in antenatal diagnosis]. AB - The authors report the case of a patient in whom an antenatal ultrasound showed a mediastinal tumor then a pleural effusion. At birth, the baby was intubated for adequate ventilation and pleural effusion was drained. Ultrasound, oesophagogram, CT scan permitted the diagnosis of tubular oesophageal duplication which was confirmed by surgery and pathologic examination. Review of the literature did not find such antenatal finding. The role of oral contraception during the first weeks of pregnancy is discussed. PMID- 3046551 TI - [Iconographic rubric. Focal nodular hyperplasia]. PMID- 3046552 TI - [Intra-hepatic calcifications diagnosed prenatally]. PMID- 3046554 TI - Differential effect of low and conventional doses of fluphenazine on schizophrenic outpatients with good or poor information-processing abilities. AB - Thirty-six stabilized schizophrenic outpatients were randomly assigned to receive either 5 or 25 mg of fluphenazine decanoate biweekly and were followed up for two years. The best and worst outcomes were found in groups with good or poor information-processing abilities (as measured by a partial-report span-of apprehension task) given the same 25-mg dose of fluphenazine decanoate. The 86% two-year survival rate of the patients with poor span performance was considerably better, while the 44% one-year and the 21% two-year survival rates of patients with good span performance were considerably lower than previously reported survival rates for schizophrenic patients receiving a conventional dose of fluphenazine. The significant correlations in a patient's span performance for periods up to one year were consistent with the hypothesis that this task taps processes associated with vulnerability to schizophrenic disorder. PMID- 3046553 TI - Clozapine for the treatment-resistant schizophrenic. A double-blind comparison with chlorpromazine. AB - The treatment of schizophrenic patients who fail to respond to adequate trials of neuroleptics is a major challenge. Clozapine, an atypical antipsychotic drug, has long been of scientific interest, but its clinical development has been delayed because of an associated risk of agranulocytosis. This report describes a multicenter clinical trial to assess clozapine's efficacy in the treatment of patients who are refractory to neuroleptics. DSM-III schizophrenics who had failed to respond to at least three different neuroleptics underwent a prospective, single-blind trial of haloperidol (mean dosage, 61 +/- 14 mg/d) for six weeks. Patients whose condition remained unimproved were then randomly assigned, in a double-blind manner, to clozapine (up to 900 mg/d) or chlorpromazine (up to 1800 mg/d) for six weeks. Two hundred sixty-eight patients were entered in the double-blind comparison. When a priori criteria were used, 30% of the clozapine-treated patients were categorized as responders compared with 4% of chlorpromazine-treated patients. Clozapine produced significantly greater improvement on the Brief Psychiatric Rating Scale, Clinical Global Impression Scale, and Nurses' Observation Scale for Inpatient Evaluation; this improvement included "negative" as well as positive symptom areas. Although no cases of agranulocytosis occurred during this relatively brief study, in our view, the apparently increased comparative risk requires that the use of clozapine be limited to selected treatment-resistant patients. PMID- 3046556 TI - [Epidemiology of breast cancer]. AB - Important facts and hypotheses of descriptive and analytic epidemiology of breast cancer are presented and discussed in connection with results of studies carried out by the authors. Leading position and further increase in breast cancer incidence in many countries are the most important point of descriptive epidemiology. In the GDR breast cancer incidence amounts to 60 per 100,000 per year. On the other hand, there are large differences in the incidence between populations, which support hypotheses about the life style, especially of diet, as a risk factor for breast cancer. The most important groups of risk factors which have been investigated are menstrual and reproductive factors, genetics, hormonal status, diet, benign breast disease, radiation and oral contraceptives. Early age at menarche and late age at menopause, nulliparity and late age at first birth, breast cancer in first-degree relatives, benign breast disease and radiation increase breast cancer risk. The results of case-control studies concerning diet are not yet convincing. A number of studies shows no overall increase of breast cancer risk. Finally, possibilities of breast cancer control are discussed. PMID- 3046555 TI - Monoclonal antibody to a human osteogenic sarcoma cell line. AB - Monoclonal antibodies against a human osteogenic sarcoma cell line were prepared by production of a somatic cell hybrids between the spleen cells from U-393OS- immunized mice and the mouse myeloma cells SP2/0. From 7 producing and well growing clones only one--B-0S12--produced antibodies, reactive preferentially with osteosarcoma cells as identified by binding second antibodies and 125I labeled Protein A. This antibody was tested against a panel of normal and tumor cell targets to determine the pattern of the antigen detected. The monoclonal antibody reacted strongly against U-3930S cells and another human sarcoma in vitro and more weakly against human fibroblasts, peripheral lymphocytes, red blood cells and was negative against mouse fibroblasts. When tested against a panel of unrelated human tumor cell lines, B-0S12 antibody was positive with melanoma cells and negative with cells from bladder, cervix and mammary carcinoma. These cross reactions suggested, that the antibody is reactive with a protein, expressed on different tumor types. This protein is not expressed on the cell surface and is probably associated with cytoskeleton, as revealed by immunofluorescence experiments. Western-blot analysis of a cytoskeletal preparation of U-3930S cells suggests, that B-0S12 antibody recognizes a protein with Mr 55 kD. Further studies are needed to characterize the molecules, carrying the epitope, identified by this monoclonal antibody. PMID- 3046557 TI - Current concepts and diagnostic evaluation of autoimmune disease. AB - This article discusses autoimmune reactions and the numerous mechanisms by which an autoimmune response may be initiated, including genetic factors, T-cell bypass mechanisms, and idiotypes. Human autoimmune diseases are classified into three main groups, ie, organ-specific, non-organ specific, and disorders with non-organ specific autoantibodies with lesions restricted to one or a few organs, that are examined in detail. General laboratory tests and interpretation of results in relation to state or treatment of the particular disease, age and sex of the patient, and the sensitivity of the test system used are reviewed. PMID- 3046558 TI - An evaluation of the Technicon H-1 automated hematology analyzer in detecting peripheral blood changes in acute inflammation. AB - The Technicon H-1 (H-1) is an automated hematology analyzer that provides a complete blood cell count, a six-part differential with absolute counts, and morphologic values with a left-shift flag (LS). To determine the sensitivity of the H-1 in detecting peripheral blood changes in acute inflammation, we first correlated the H-1 LS with band counts on the Hematrak 590 (H590), an automated digital image processor differential system. The H-1 sensitivity to H590 band counts above 11% was 76%, specificity was 82%, and efficiency was 80%. Each semiquantitative LS (1+, 2+, 3+), as well as a new factor, lobularity index, was correlated with the actual H590 band count. There was a definite direct proportional relationship between each semiquantitative LS and the mean band count. However, the wide overlaps of band count ranges corresponding to each semiquantitative flag rendered semiquantitation of limited value. Forty cases with the clinical diagnosis of acute appendicitis were similarly studied preoperatively. Thirty-three cases histologically showed acute inflammation. On the H-1, 79% (sensitivity) had LS flags, 88% had absolute neutrophilia (greater than 8 x 10(9)/L), 82% had relative neutrophilia (greater than 75%), and 91% had leukocytosis (greater than 10.5 x 10(9)/L). In comparison, sensitivities on the H590 were 70% for band counts above 11%, 82% for relative neutrophilia, and 85% for absolute neutrophilia. This study shows that the H-1 is at least as sensitive as the H590 to peripheral blood changes that indicate acute inflammation. PMID- 3046559 TI - Interference in immunoenzymometric assays caused by IgM anti-mouse IgG antibodies. AB - Serum samples from a female patient gave falsely elevated results in nine of ten immunoenzymometric assays (IEMAs) that used mouse monoclonal antibodies specific for human chorionic gonadotropin (n = 8), thyrotropin, or creatine kinase-MB isozyme. In contrast, normal results were obtained in five radioimmunoassays that used either mouse monoclonal antibodies or antisera specific for human chorionic gonadotropin (n = 3) or thyrotropin (n = 2). Incubation of her serum with IgG from different species or with F(ab)'2 from mouse IgG prior to IEMA showed that the interference was markedly inhibited by mouse IgG, indicating an antibody specific for the Fc portion of mouse IgG. The interfering activity was bound to and eluted from a column containing Protein A-Sepharose CL-4B. Fractionation of the eluted protein over another column containing Sephacryl S300 showed the activity was enriched in the first protein peak, which contained predominantly IgM. A model is proposed to explain how IgM anti-mouse IgG antibody selectively interferes in IEMAs that use mouse monoclonal antibodies. PMID- 3046560 TI - Medical advances during the Civil War. AB - The contributions to medical care that developed during the Civil War have not been fully appreciated, probably because the quality of care administered was compared against modern standards rather than the standards of the time. The specific accomplishments that constituted major advances were as follows. 1. Accumulation of adequate records and detailed reports for the first time permitted a complete military medical history. This led to the publication of the Medical and Surgical History of the War of the Rebellion, which was identified in Europe as the first major academic accomplishment by US medicine. 2. Development of a system of managing mass casualties, including aid stations, field hospitals, and general hospitals, set the pattern for management of the wounded in World War I, World War II, and the Korean War. 3. The pavilion-style general hospitals, which were well ventilated and clean, were copied in the design of large civilian hospitals over the next 75 years. 4. The importance of immediate, definitive treatment of wounds and fractures was demonstrated and it was shown that major operative procedures, such as amputation, were optimally carried out in the first 24 hours after wounding. 5. The importance of sanitation and hygiene in preventing infection, disease, and death among the troops in the field was demonstrated. 6. Female nurses were introduced to hospital care and Catholic orders entered the hospital business. 7. The experience and training of thousands of physicians were upgraded and they were introduced to new ideas and standards of care. These included familiarity with prevention and treatment of infectious disease, with anesthetic agents, and with surgical principles that rapidly advanced the overall quality of American medical practice. 8. The Sanitary Commission was formed, a civilian-organized soldier's relief society that set the pattern for the development of the American Red Cross. PMID- 3046561 TI - Presentation of the Murphy gavel. PMID- 3046562 TI - Porcine parvovirus: replication in and inhibition of selected cellular functions of swine alveolar macrophages and peripheral blood lymphocytes. AB - The ability of four isolates of Porcine Parvovirus (NADL-8, NADL-2, KBSH, and Kresse) to replicate in and affect the functions of swine peripheral blood lymphocytes and alveolar macrophages was studied in vitro. V-strand and C-strand viral DNA was present in both concanavalin A- and non-treated lymphocytes as well as alveolar macrophages following infection with all four isolates. Indirect fluorescent antibody assays on swine testis cells, inoculated with cell lysates of NADL-8-infected peripheral blood lymphocytes (both concanavalin A- and non treated) and alveolar macrophages, indicated that these immune cells supported the production of progeny virus. The quantity of viral DNA and progeny virus was dependent upon the multiplicity of infection and length of time following infection. Infection of lymphocytes and alveolar macrophages with PPV was associated with a decrease in cell viability. Peripheral blood mononuclear cells and alveolar macrophages infected with any of the four isolates demonstrated reduced lymphocyte blastogenesis and non-Fc-mediated alveolar macrophage phagocytosis, respectively. PMID- 3046564 TI - [Identification of autoantibody-inductive B cell-stimulation factor and gene regulation of an MHC-linked B cell response]. PMID- 3046563 TI - Antibody response to wild rubella virus structural proteins following immunization with RA 27/3 live attenuated vaccine. AB - Antibody response to individual structural proteins (E1, E2, and C) of the M-33 wild rubella virus strain was assayed by an immunoblot technique in 12 girls, following immunization with RA 27/3 live attenuated rubella vaccine. Of the 12 immunized subjects, before vaccination 9 had no demonstrable rubella specific antibodies while the remaining 3 had a low level of rubella specific antibodies, reacting only with the E1 protein. At one month after vaccination all the immunized subjects presented anti-E1, anti-E2, and anti-C specific antibodies. However, at 1-2 weeks after vaccination the 9 girls who were seronegative before immunization still had no detectable antibodies to any of the rubella virus structural proteins, while the 3 subjects whose preimmunization sera had reacted with the E1 protein presented an accelerated immune response, showing anti-E2 and anti-C specific antibodies and a more intensely marked anti-E1 specific band. PMID- 3046565 TI - [Drug interactions between quinolone antibodies and theophylline]. PMID- 3046567 TI - [Architectonics of the vascular bed of the fetal portion of the mature human placenta (based on data of scanning electron microscopy)]. AB - Peculiarities of the microangioarchitectonics in the trunks, intermediate and terminal villi of the placenta have been described at a noncomplicated pregnancy. Signs of neovasculogenesis are demonstrated in the microcirculatory bed of the terminal villi. Possible connection between reduction of the paravasal capillary network, as the intermediate villi become mature, and neovasculogenesis is discussed. PMID- 3046566 TI - [Evaluation of a new kit for measuring theophylline (AccuLevel)]. PMID- 3046568 TI - [Principles and technic of the topometry of the calyx-pelvis structure of the human kidney]. PMID- 3046569 TI - [Macro- and microscopic characteristics of the luminal surface of the small intestine]. PMID- 3046570 TI - [Sensitive innervation of the skin of the concha auriculae]. AB - By means of the retrograde transport of horseradish peroxidase (HP) method sensitive innervation of the rabbit concha auriculae skin has been studied. For the investigation the skin area 2 X 2 sm large has been chosen on the internal surface of the right concha auriculae 6 sm below the upper edge of the ear. The HP solution is injected in the skin areas 5-6 mm large with normal electrical permeability and with decreased electrical resistance to the constant electrical current. In the first case innervation is ensured only by ipsilateral sensitive neurons of the spinal nodes (SN) CIII. Small neurons make 75% of the total amount of the labelled cells. In the second case about 98% of the skin areas are innervated by the neurons of the SN CIII and about 2% by the neurons of the SN CII. Small and middle neurons make 60% and 38%, respectively. Single labelled cells are revealed in the SN CIV. The HP-positive neurons, innervating the skin areas with an increased electrical permeability nearly 3 times exceed the neurons, projecting on the skin areas with normal electrical permeability. The large labelled neurons make 2% in both cases. Localization of the HP-positive cells in the SN is diffuse, structural compositions of the neurons are not revealed. In the trigeminal node labelled neurons are not found. PMID- 3046572 TI - [Ultrastructure of the inner surface of the aorta of mature and old animals]. AB - In the experiments, performed on 12 white rats and 8 rabbits, by means of scanning electron microscopy of the native preparations and in a number of cases with use of silver nitrate impregnation, the internal surface structure has been studied in the aortal membrane of mature and old animals. At ageing the integrity and continuity of the endothelial monolayer is preserved, on the surface local intimal pits, craters and microdefects appear, adhesiveness of endothelium to leucocytes increases. Orientation of the intimal folds is disturbed. The type of the senescent remodeling in the endothelial layer revealed predisposes to development of atherosclerosis. PMID- 3046571 TI - [Immunohistochemical detection of lactoferrin of human milk in the tissues of the human and the fetus]. AB - By means of the indirect immunoperoxidase method in deparaffinized slices a comparative investigation on distribution of the female milk lactoferrin (LF) in tissues of the mature person and in the fetus has been performed. LF is revealed in cells of the neutrophil line of the mature person and of the fetus and in the secretory epithelium of some organs of the mature person (in the mammary, submandibular and parotid salivary glands, in the bronchial mucous membrane glands in the fundal and pyloric glands of the stomach). In all the cases investigated LF is revealed in the cells producing serous secrete: in the cells of the serous terminal parts and in the serous semilunar mixed terminal parts of the salivary glands, in the serous cells of the bronchial glands and in the chief cells of the gastric mucous membrane glands. In the fetal secretory epithelium of the organs LF is not found. As LF is revealed in the secretory epithelium of the mature person and is not revealed in the corresponding epithelium of the fetus, it should be considered as a marker of the cells, that reach certain degree of differentiation. PMID- 3046573 TI - [Regional characteristics of the organization of the aortic endothelium (a quantitative analysis)]. AB - By means of SEM-analysis in 110 microphotos (from 14 aortas of white rats) with the aim to estimate heteromorphism of the thoracic part endothelium, quantitative characteristics of endotheliocytes have been studied in the ventral and dorsal surfaces, and also around the ostia of the intercostal arteries. For the quantitative analysis organization of the lines of the interendotheliocyte borders is taken into account. Endotheliocytes around the ostium are more elongated but occupy less area and have less straight contours in comparison with cells in other regions; they have also greater variability. Essential differences in arranging the endothelial cells into the layer on the ventral and dorsal surfaces of the aorta are revealed. PMID- 3046574 TI - [Changes in the blood vessels of the lungs under physical loads at high altitude]. AB - Changes of the pulmonary artery branches in rats at the state of relative physiological rest undergo two stages during the process of individual adaptation for 60 days. The first is characterized with generalized decrease of the lumen and with thickening of the vascular wall as a result of reflectory spasm and edematous swelling of the arteries, the second--with a relative dilatation of the lumen and with decreasing thickness of the vascular wall, as a result of decreasing reflectory spasm and edematous phenomena. The physical load increases the organism's elevated oxygen consumption under conditions of high-altitude, which is increased by itself; this intensifies the function of the pulmonary blood bed. In combination with the edematous phenomena, the wall of the pulmonary artery becomes sharply thickened in the first stage. In the second stage the tissue mechanisms are switched on for satisfaction of oxygen consumption. Therefore, the load on the right part of the heart and on the pulmonary vessels decreases edematous phenomena, the thickness of the wall in the pulmonary artery branches decline, in comparison with the data of the first stage. The thickness of the wall in the first stage is combined with a manifested constriction of the lumen, and the relative thinning of the vascular wall--with dilatation. PMID- 3046575 TI - [Ultrastructure of the cells of the neural crest]. AB - Ultrastructure of the neural crest cells (NCC) has been studied in 9-day-old white rat embryos. Mechanisms, determining the amount of the NCC that get out of the neuroepithelium of the nervous tube have been discussed. By means of ruthenium red components of the extracellular matrix are demonstrated in the area where the NCC get out and in the ventromedial pathway of migration. Certain peculiarities of ruthenium red fixation by the cells are presented. PMID- 3046576 TI - [Methodology of morphological research based on material from the journal "Arkhiv anatomii, gistologii i embriologii" 1982-1986]. PMID- 3046578 TI - Theodore Herpin. A mid-19th century view on epilepsy. PMID- 3046577 TI - [Clinico-morphological aspects of placental insufficiency]. AB - The paper is concerned with the problems of placental insufficiency diagnosis, pathogenesis and potential effect on the fetus. The diagnosis involves the comparison of the compensatory and adjustment reaction to the number and pattern of pathological placental alterations in line with clinical and laboratory findings. The author employed the methods of macro-, microstereometry, electron microscopy, histochemistry, immunofluorescence. The essential role in placental insufficiency pathogenesis is attributed to the failure of regulatory metabolic mechanisms. This permits further investigation with employment of membrane pathology and biochemical methods of investigation. PMID- 3046579 TI - Ceftazidime-induced encephalopathy in a patient with renal impairment. PMID- 3046580 TI - Lateral thalamic infarcts. AB - A patient with occlusion of the proximal posterior cerebral artery (PCA), a lateral thalamic infarct, and hemisensory loss later developed hemianopia and hemiparesis and had extensive PCA territory infarction in the midbrain, the lateral portion of the thalamus, and the occipital lobe noted at necropsy. Two other patients had lateral thalamic infarcts on computed tomography, normal angiographic findings, and presumed thalamogeniculate artery branch occlusion. There are three clinical syndromes associated with lateral thalamic infarction: (1) hemisensory loss, hemiataxia, and involuntary movements; (2) pure sensory stroke; and (3) sensory-motor stroke. Ataxia, adventitious movements, and sensory loss are due to infarction of the lateral, posterolateral, and posteromedial ventral nuclei caused by occlusion of the PCA proximal to the thalamogeniculate artery branches or by occlusion of large thalamogeniculate arteries. Pure sensory and sensory-motor strokes are due to smaller infarcts in the posterolateral posteromedial ventral complex and adjacent internal capsule caused by occlusion of penetrating artery branches of the thalamogeniculate arteries. PMID- 3046581 TI - Suturing of stents after dacryocystorhinostomy. PMID- 3046582 TI - Acanthamoeba keratitis. Potential role for topical clotrimazole in combination chemotherapy. AB - Clotrimazole is an antifungal agent that has been shown to have excellent in vitro activity against most strains of Acanthamoeba. We encountered four patients who developed Acanthamoeba keratitis while wearing contact lenses that had been stored in homemade saline. Their medical treatment regimens included the use of topical 1% clotrimazole. In two patients in whom conventional therapy failed, clotrimazole was successful in controlling recurrent infection following penetrating keratoplasty. Two other patients were treated with clotrimazole as well as propamidine isethionate and neomycin sulfate-polymyxin B sulfate gramicidin from the outset, and had an excellent response to medical therapy. In those patients who found the commercially available cream uncomfortable, a 1% clotrimazole suspension formulated in artificial tears was used and found to be well tolerated. PMID- 3046584 TI - Clinical evaluation of the Cavitron Biotronics tonometer. AB - The Mackay-Marg tonometer has been shown to be an accurate tonometer for the measurement of intraocular pressure in eyes with both normal and diseased corneas. This tonometer is no longer manufactured and has been replaced by the Cavitron Biotronics tonometer that works on the same principle but utilizes a different recording display. The Cavitron Biotronics tonometer's accuracy was compared with that of the Goldmann applanation tonometer in 75 eyes. The Cavitron Biotronics tonometer is not as accurate as the original Mackay-Marg tonometer for individual readings. However, if four readings are taken, the value of the Cavitron Biotronics tonometer that correlated best with Goldmann applanation tonometry was the one reading of the four with the highest value. Using this one value, the Cavitron Biotronics instrument underestimates the true intraocular pressure below 27 mm Hg and overestimates it above this value. PMID- 3046583 TI - Elevated corneal epithelial lines in Acanthamoeba keratitis. AB - Elevated corneal epithelial lines are another clinical sign in Acanthamoeba corneal infection. In this report, one patient wore extended wear soft contact lenses, and another wore daily wear soft contact lenses. Both patients used distilled water and salt tablets in their lens care. Histopathologic examination of these lines revealed trophozoites and cysts. In one of the patients following penetrating keratoplasty, Acanthamoeba castellani and Acanthamoeba polyphaga were cultured by impression cytology of an epithelial line, as well as from the bulbar and tarsal conjunctiva. In the other patient who did not undergo penetrating keratoplasty, these lines appeared in the cornea one month after initial symptoms. PMID- 3046585 TI - Penetrating keratoplasty after epikeratophakia for keratoconus. AB - Seven patients underwent penetrating keratoplasty after epikeratophakia for keratoconus. All seven patients achieved clear grafts and 20/40 or better best corrected visual acuity after penetrating keratoplasty. Three of the seven patients had one or more episodes of rejection after penetrating keratoplasty; all were treated successfully. No grafts were lost. The results in terms of graft clarity and visual acuity are comparable with those in patients undergoing penetrating keratoplasty for keratoconus with no previous ocular surgery. Whether the relatively high rate of rejection episodes (three [43%]) seen in this small number of patients indicates some relationship between previous epikeratophakia and subsequent rejection after penetrating keratoplasty remains to be seen. PMID- 3046586 TI - The findings of standardized echography for choroidal folds. AB - Twenty-four patients (31 eyes) with choroidal folds unassociated with orbital tumors were evaluated with standardized echography. Hypermetropia was the most commonly associated finding (eight eyes); in ten eyes, no consistent abnormal findings could be established. Among the less common causes were swelling of the optic nerve proper or the perineural sheaths and thickened extraocular muscles. Standardized echography demonstrated ocular changes, orbital changes, or both, in all but two patients (two eyes) with idiopathic folds. The most frequent findings were flattening of the posterior ocular wall (18 eyes), thickening of the retinochoroid layer (12 eyes), and distention of the optic nerve sheaths (eight eyes). While fluorescein angiography is well established as the preferred method of demonstrating choroidal folds, standardized echography may now be used to delineate the often subtle associated ocular and orbital findings. PMID- 3046587 TI - The response of diabetic retinopathy to 41 months of multiple insulin injections, insulin pumps, and conventional insulin therapy. AB - Forty-five diabetic patients were randomized and treated with continuous subcutaneous insulin infusion (CSII), multiple insulin injections (MI), or conventional insulin treatment (CIT) for 41 months. Near-normoglycemia was obtained with CSII and MI but not with CIT. A transient increase in microaneurysms and hemorrhages was seen at three months in CSII-treated patients. After 41 months, a moderate progression in microaneurysms and hemorrhages was registered, as assessed from fundus photographs, in all treatment groups. Fluorescein angiograms indicated a tendency (not statistically significant) to retarded progression of retinopathy in MI- and CSII-treated patients compared with CIT-treated patients. Soft exudates developed after three to six months of rapid tightening of metabolic control in 50% of patients on CSII and MI regimens. Those patients who had soft exudates had a slower progression of retinopathy three years later than those who did not develop soft exudates. Transient progression of retinopathy may be related to fluctuations in blood glucose levels, although a favorable effect of long-term improved metabolic control was not documented. PMID- 3046588 TI - Reliability indexes of automated perimetric tests. AB - The "reliability" of a subject's automated perimetric test result is generally assessed by three measures: fixation loss and false-positive and false-negative rates. These reliability criteria were examined for 76 glaucomatous and 248 normal subjects who underwent visual field testing (C-30-2 program; Humphrey Visual Field Analyzer, Allergan Humphrey, San Leandro, Calif). Of the examination results, 45% in glaucomatous subjects and 30% in normal controls were considered unreliable with the use of the manufacturer's reliability criteria. Most test results were unreliable because they failed to meet the criterion for fixation loss. The greater rejection rate among glaucomatous subjects was entirely due to their higher rate of false-negative responses. Factors such as age, pupil diameter, and visual acuity did not explain the difference between the false negative rates of glaucomatous patients and normal subjects. PMID- 3046589 TI - Delayed primary wound closure. Use to prevent implant extrusion following evisceration for endophthalmitis. AB - We used delayed primary wound closure in three cases of bacterial endophthalmitis to minimize the risk of implant extrusion following evisceration. In a fourth case, the wound was closed primarily, but wound dehiscence and implant extrusion occurred six weeks postoperatively, and reoperation was required. The advantages of delayed primary wound closure following evisceration of the globe for endophthalmitis include rapid removal of the intraocular abscess, continued drainage and mechanical debridement of the infected scleral pouch, and the development of granulation tissue resistant to bacterial growth at the wound margins prior to wound closure. These factors are important in reducing the risk of implant extrusion following evisceration of the globe for endophthalmitis. PMID- 3046590 TI - Leg ulcers. PMID- 3046591 TI - The adult health screen in general practice. PMID- 3046592 TI - The use of pre-operative scan prior to neck exploration for primary hyperparathyroidism. AB - A group of patients with diagnosed primary hyperparathyroidism (PHP) in Auckland between 1982 and 1986 is reviewed. Of the 119 patients, 55 had pre-operative scanning, 36 had no scanning prior to surgery and 29 were managed conservatively. Of the 52 patients who had pre-operative localization with ultrasound scanning, only 27 (52%) had their adenoma correctly predicted upon neck exploration. Of the 14 patients investigated with thallium-201 and technetium-99m (T1/Tc) subtraction scanning, 10 (71%) had their adenoma positively identified in the predicted locations, whereas the cause of the parathyroid pathology was correctly identified in 33 of the 36 patients (92%) who had surgery alone with no pre operative scanning. In 11 patients both ultrasound and T1/Tc subtraction scanning were employed. In eight patients the results of the two scanning modalities agreed, and in seven of these eight patients the adenoma was correctly predicted (six single adenomas and one with double adenoma). In the three patients in whom the results of the two modalities differed, the T1/Tc subtraction scanning correctly predicted the site of the adenoma in two patients and in the third patient (with a small 223 mg adenoma) both modalities were incorrect. Of the six patients with histologically proven parathyroid hyperplasia, only three had pre operative localization with ultrasound alone, and all three had incorrect predictions (one false positive for adenoma, and two false negative scans). Overall the results cast doubt over the usefulness of pre-operative scanning as a routine investigation prior to initial neck exploration for primary hyperparathyroidism.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3046594 TI - The University of Melbourne/Nucleus cochlear prosthesis. PMID- 3046595 TI - The Surgical Research Society of Australasia--a silver jubilee. PMID- 3046593 TI - Melbourne trial of adjuvant immunotherapy in operable large bowel cancer. AB - A controlled randomized clinical trial was undertaken to assess the ability of combined non-specific and specific immunotherapy to alter the disease-free interval and overall survival of patients with Stage B or C large bowel cancer. The immunotherapy consisted of a 2 year programme of vaccinations with BCG and neuraminidase-treated autologous tumour cells. Three hundred and one patients entered the trial. At 5 years of follow-up there is no evidence that this form of immunotherapy can alter either the disease-free interval or survival in this group of patients. PMID- 3046596 TI - Aerospace Medical Association. Past presidents. PMID- 3046597 TI - Aerospace Medical Association. Directory of members. PMID- 3046599 TI - The use of immunostaining to quantify x-ray and heat-induced multiple microtubule organizing centers in normal and transformed cells. AB - Microtubule organizing centers (MTOC) in control, irradiated and heated C3H 10T1/2 mouse embryo cells and two radiation-transformed sublines, R1 and R25, were made visible by indirect immunofluorescence using antibody against tubulin. The MTOC were reformed by 5-min incubation in fresh medium after the microtubules were depolymerized with nocodazole. The R1 line had a different distribution of MTOC/cell than the parent 10T1/2 line or R25, which had similar distributions. After irradiation, multiple MTOC appeared in the normal and radiation-transformed cells irradiated to 10 Gy and incubated for 24 or 48 h. The multiple foci of microtubule reformation in the irradiated cells indicate that radiation damage is expressed in structural elements in the cytoplasm. After heat treatment of the three cell lines (43 degrees C for 93 min and 45 degrees C for 25 min), the MTOC were disrupted and many cells did not have visible organizing centers at 24 or 48 h, while others had a large number of small centers of microtubule reformation. The distribution of MTOC/cell seen in R25 cells after the treatment had similar patterns to those of the 10T1/2 line rather than to those of the other radiation transformed line, R1. Thus, the radiation or heat response seen in the MTOC is not dependent upon cell transformation. PMID- 3046598 TI - Histological and immunohistochemical features in chronic delta hepatitis. AB - Forty-six liver biopsies from HBsAg Delta/anti-Delta serum positive patients with chronic hepatitis were compared with liver specimens from HBsAg positive Delta/antiDelta negative patients. The results indicate more severe histologic damage and inflammation among subjects who are serum-positive to the Delta system. Specific virus antigen was found in liver cell nuclei of almost all the Delta IR patients (93.5%). The rate of diffusion was directly proportional to the severity of histological lesions, which is in linea with the direct cytopathic effect of the Delta virus as ascertained in various other studies. PMID- 3046600 TI - Development of insulin-sensitivity at weaning in the rat. Role of the nutritional transition. AB - This study was undertaken to determine the factors involved in the development of insulin-sensitivity at weaning. Glucose kinetics were studied in suckling rats and in rats weaned on to a high-carbohydrate (HC) or a high-fat (HF) diet, in the basal state and during euglycaemic-hyperinsulinaemic-clamp studies. These studies were coupled with the 2-deoxyglucose technique, allowing a measure of glucose utilization by individual tissues. In the basal state, the glycaemia was higher in HF-weaned rats (124 +/- 4 mg/dl) than in suckling (109 +/- 1 mg/dl) and HC weaned rats (101 +/- 3 mg/dl). Glucose turnover rates were similar in the three groups of animals (14 mg/min per kg). Nevertheless, basal metabolic glucose clearance rate was 20% lower in HF-weaned rats than in the other groups. During the euglycaemic-hyperinsulinaemic experiments, hepatic glucose production was suppressed by 90% in HC-weaned rats, whereas it remained at 40% of basal value in suckling and HF-weaned rats, indicating an insulin resistance of liver of these animals. Glucose clearance rate during the clamp was 18.3 +/- 0.9 ml/min per kg in suckling rats, whereas it was 35.3 +/- 1.2 ml/min per kg in HC-weaned rats and 27.8 +/- 1.1 ml/min per kg in HF-weaned rats, indicating an insulin resistance of glucose utilization in suckling, and to a lower extent, in HF-weaned rats. The deoxyglucose technique showed that peripheral insulin resistance was localized in muscles and white adipose tissue of suckling and HF-weaned rats. These results indicate that the switch from milk to a HC diet is an important determinant of the development of insulin-sensitivity at weaning in the rat. PMID- 3046602 TI - Regulation of the breakdown rates of biotin-containing proteins in Swiss 3T3-L1 cells. AB - 1. Degradation rate constants for individual biotin-labelled proteins were measured in Swiss 3T3-L1 adipocytes that had been incubated with inhibitors of autophagy or of lysosomal proteolysis. 2. Inhibitory effects produced by 10 mM-3 methyladenine and a combination of 5 mM-NH4Cl and leupeptin (50 micrograms/ml) were approximately equal. The inclusion of NH4Cl did not significantly enhance the responses to 3-methyladenine, suggesting that autophagy was already maximally inhibited. 3. The extent of inhibition by 3-methyladenine or by the NH4Cl/leupeptin mixture was similar for the cytosolic enzyme acetyl-CoA carboxylase and for the three mitochondrial carboxylases. This inhibition averaged 50%. The breakdown rate of a more-stable 38 kDa biotin-containing mitochondrial protein was more responsive to the inhibitory agents. These results are best explained by mitochondrial proteolysis occurring via a combination of the degradation of whole mitochondria within autophagic vacuoles, supplemented by the selective intramitochondrial breakdown of more labile proteins. 4. A number of intermediate products in the degradation of biotin-containing proteins were detected. Differences in the patterns of radioactivity between these peptides after incubation of cells in the presence of inhibitors of the breakdown process provided evidence that some peptides were produced before autophagy, others as a result of intralysosomal inhibition, while at least one was associated with intramitochondrial proteolysis. PMID- 3046601 TI - The response of adipocyte glucose metabolism and fatty acid release to adenosine deaminase, insulin and perifusion. Investigation of intermediary metabolism by perifusion. AB - Adipocytes incubated with adenosine deaminase (ADA) showed: (1) increased amounts of fatty acids in the medium; (2) increased glucose incorporation into acylglycerol glycerol; (3) decreased glucose incorporation into acylglycerol fatty acids; (4) a co-ordinate decrease in the sensitivity of lipolysis and glucose incorporation into acylglycerol to insulin; (5) similar effects on glucose incorporations in perifused and normal incubations. The decrease in fatty acid synthesis by perfusion was found to be dependent on the presence of insulin or fatty acids, and independent of the effects of ADA. The significance of the effects of perifusion, ADA and insulin are discussed in relation to effects of fatty acids. PMID- 3046603 TI - A heparin-binding protein involved in inhibition of smooth-muscle cell proliferation. AB - A heparin-binding protein was isolated from bovine uteri and purified to homogeneity. This protein appears as a double band of approx. 78 kDa in SDS/polyacrylamide-gel electrophoresis and has an isoelectric point of 5.2. The binding of heparin to this protein is saturable. No other glycosaminoglycan from mammalian tissue, such as hyaluronic acid, chondroitin sulphate, dermatan sulphate or keratan sulphate, binds to the 78 kDa protein. Dextran sulphate binds in a non-saturable fashion. Certain heparan sulphate polysaccharide structures are required for binding to the 78 kDa protein. Some proteoheparan sulphates, such as endothelial cell-surface proteoheparan sulphate, show only weak interaction with the 78 kDa protein in contrast with a basement-membrane proteoheparan sulphate from HR-9 cells. Antibodies against the 78 kDa protein inhibit binding of proteoheparan [35S]sulphate from basement membranes to smooth muscle cells. Conventional antibodies, Fab fragments and some monoclonal antibodies, inhibit smooth-muscle cell proliferation in a similar range as that observed for heparin. The protein was detected in a variety of tissues and cells but not in blood cells. A possible role of this protein as a receptor for heparin or heparan sulphate and its function in the control of the arterial wall structure are discussed. PMID- 3046604 TI - Isolation and expression of a pea vicilin cDNA in the yeast Saccharomyces cerevisiae. AB - A cDNA clone containing the complete coding sequence for vicilin from pea (Pisum sativum L.) was isolated. It specifies a 50,000-Mr protein that in pea is neither post-translationally processed nor glycosylated. The cDNA clone was expressed in yeast from a 2 micron plasmid by using the yeast phosphoglycerate kinase promoter and initiator codon. The resultant fusion protein, which contains the first 16 amino acid residues of phosphoglycerate kinase in addition to the vicilin sequence, was purified and subsequently characterized. It has slightly slower mobility on SDS/polyacrylamide-gel electrophoresis than standard pea vicilin and forms a mixture of multimers, some of which resemble the native protein. PMID- 3046605 TI - Phorbol ester inhibits arginine vasopressin activation of phospholipase C and promotes contraction of, and prostaglandin production by, cultured mesangial cells. AB - We have previously shown that arginine vasopressin (AVP) causes a rapid (5-10 min) contractile response in cultured mesangial cells plated onto slippery substrata such as poly(hydroxyethyl methacrylate)-coated dishes. This contraction is associated with an increase in the levels of inositol trisphosphate (InsP3), diacylglycerol and prostaglandin E2 (PGE2). We now report that agents which are known to activate protein kinase C, i.e. phorbol 12-myristate 13-acetate (PMA) and oleolylacetylglycerol (OAG), also contract mesangial cells; however, the contractile response is slow to develop (15-30 min). The inactive phorbol ester, 4 alpha -phorbol 12,13-didecanoate, did not elicit contraction. PMA and OAG did not increase InsP3 release in mesangial cells. However, pretreatment of mesangial cells with PMA inhibited the formation of InsP3. This inhibition could not be explained by a reduction in AVP binding since PMA treatment did not influence the number or affinity of [3H]AVP binding sites in intact cells. PMA alone stimulated PGE2 production in mesangial cells to a degree similar to AVP. Contrary to what was seen with InsP3, pretreatment of cells with PMA before AVP had an additive effect on arachidonic acid release and PGE2 production. Thus, there is an apparent dissociation of phospholipase C activity from that of phospholipase A2. PMID- 3046607 TI - Iron K-edge X-ray absorption spectroscopy of the iron-molybdenum cofactor of nitrogenase from Klebsiella pneumoniae. AB - Iron K-edge X-ray absorption data for the iron-molybdenum cofactor ('FeMoco') from Klebsiella pneumoniae reported here provide the first evidence for long range structural order in the cofactor [Fe...Fe(Mo) = 0.368 nm in addition to Fe...S = 0.22 nm and Fe...Fe(Mo) = 0.27 nm] and, in contrast with previously published data [Antonio, Teo, Orme-Johnson, Nelson, Groh, Lindahl, Kauzlarich & Averill (1982) J. Am. Chem. Soc. 104, 4703-4705], indicate that most of the iron centres are not co-ordinated to light (oxygen, nitrogen) atoms. This demonstrates that presently available chemical models for FeMoco are inadequate. PMID- 3046608 TI - Release of copper from yeast copper-thionein after S-alkylation of copper thiolate clusters. AB - Our knowledge on the release of copper from Cu-thionein in biological systems is limited. Other than oxidative cleavage or direct transfer, the possibility of an alkylation mechanism seemed attractive. Iodoacetamide and methyl methanesulphonate were successfully employed to alkylate the Cu-thiolate sulphur atom of homogeneous Cu(I)-thionein from yeast. The alkylation caused a weakening of the Cu-S bonding, which led to the release of copper. After equilibrium dialysis a proportion of the released copper was found in the dialysis buffer. When iodoacetamide was used carboxymethylcysteine was detected in the protein hydrolysate. A 10-fold molar excess over cysteine was sufficient for complete alkylation, which could be conveniently monitored by c.d. at 328 and 359 nm. The reaction proceeded under both aerobic and anaerobic conditions. E.p.r. measurements of Cu2+ revealed unequivocally the complete cleavage of the Cu thiolate bonding in less than 5 h. It is possible that this mode of copper release might be of relevance to the molecular transport of this biochemically important transition metal. PMID- 3046609 TI - Early effects of Escherichia coli endotoxin infusion on vasopressin-stimulated breakdown and metabolism of inositol lipids in rat hepatocytes. AB - The turnover of vasopressin-stimulated 32P-phosphoinositides and 32P-phosphatidic acid and accumulation of [2-3H]-inositol phosphates were examined in hepatocytes from rats infused i.v. with saline and E. coli endotoxin for 3 hrs. Within 60s of VP stimulation the decrease in phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate labeling as well as the increased uptake of 32P into phosphatidic acid were similar in both groups. However, at a later time (300s) the 32P-phosphatidylinositol turnover was greatly decreased concomitantly with a higher labeling of phosphatidic acid. The accumulation of [2-3H]-inositol phosphates in ET-cells was significantly decreased both at 30s and 600s after VP addition. The distribution of [2-3H]-inositol labeling accumulated in the different inositol phosphate fractions over the first 30s of VP stimulation showed a tendency to lower accumulation of inositol trisphosphate, and a significantly lower accumulation of inositol bisphosphate simultaneously with a higher labeling of the inositol tetrakisphosphate fraction. These observations reflect an early effect of ET-infusion on VP-stimulated inositol lipid turnover and on the subsequent metabolism of the released inositol phosphates. PMID- 3046610 TI - The ontogeny of the rabbit hepatic glucose transporter. AB - We delineated and characterized the fetal hepatic glucose transporter in the rabbit. Employing the 2-deoxy-D-glucose displaceable 3H-cytochalasin B binding assay we estimated the number and Kd of the GT per mg of liver protein. A gradual increase in the number was observed during development, the fetus (23.8 +/- 2.04 pmoles/mg) expressing a lesser amount when compared to the neonate (59.5 +/- 17 pmoles/mg; p less than 0.05) and adult (142 +/- 11 pmoles/mg; p less than 0.05). On the other hand the affinity of the glucose transporter was higher in the fetus (Kd 287 +/- 81 nM) when compared to either the neonate (988 +/- 222 nM, p less than 0.05) or the adult (706 +/- 101 nM, p less than 0.05). We conclude that the fetal hepatic GT is more efficient secondary to a higher affinity for glucose. PMID- 3046606 TI - Proteoglycan-fibrillar collagen interactions. PMID- 3046611 TI - Existence of anti-thyroglobulin IgG in healthy subjects. AB - An autoantibody, anti-thyroglobulin IgG, was detected in a large proportion of healthy subjects. Sera were collected from 232 healthy subjects aged 7-83 yr, who had no apparent symptoms with normal serum levels of thyroid-stimulating hormone, confirming the absence of Graves' disease and chronic thyroiditis. Anti thyroglobulin IgG in serum was measured by a novel enzyme immunoassay, the principle of which has been shown to provide 3,000 to 10,000-fold higher sensitivity than the conventional methods. Anti-thyroglobulin IgG was demonstrated in 38% of the healthy subjects (15% of those aged 7-19 yr and 69% of those aged 20-39 yr), and the serum concentration of anti-thyroglobulin IgG was assessed to be 2 micrograms/l - 38 mg/l. PMID- 3046612 TI - Effect of fibroblast-derived differentiation inducing factor on the differentiation of human monocytoid and myeloid leukemia cell lines. AB - A fibroblast-derived differentiation inducing factor (F-DIF) purified from medium conditioned by a human fibroblast cell line (WI-26VA4) induced differentiation of human monocytic leukemia cell lines (U-937, THP-1) into cells with macrophage characteristics. F-DIF alone induced the differentiation of ML-1 cells only marginally, but it synergistically increased the differentiation when combined with TNF. Interferon-gamma, tumor necrosis factor, GM-CSF, interleukin-1 and interlukin-4 synergistically enhanced the differentiation of U-937 cells when combined with F-DIF. PMID- 3046613 TI - 12-O-tetradecanoylphorbol-13-acetate activates phosphatidylethanol and phosphatidylglycerol synthesis by phospholipase D in cell lysates. AB - A cell-free system for the synthesis of phosphatidylalcohols was developed in sonicates of HL-60 cells. With [32P]phosphatidylcholine as the exogenous substrate, both phosphatidylethanol and phosphatidylglycerol were formed through a phospholipase D-catalyzed transphosphatidylation of ethanol and glycerol, respectively. The transphosphatidylation by phospholipase D was stimulated in vitro by 12-O-tetradecanoylphorbol-13-acetate (TPA) and required the addition of ATP for an optimal response. GTP-gamma-S, an activator of G protein systems, also stimulated the process by an independent mechanism. It is postulated that the stimulation of phospholipid metabolism through phospholipase D activation represents an important mechanism whereby TPA might modulate intracellular signal generating systems or influence the activity of membrane-bound proteins by altering their lipid environment. PMID- 3046614 TI - Host-cell attachment by Trypanosoma cruzi: identification of an adhesion molecule. AB - We have identified an 83 kDa surface glycoprotein in T. cruzi trypomastigotes which specifically binds to rat heart myoblasts. The binding of this molecule to myoblasts is inhibited by excess unlabeled material and saturable. Antibodies against the cell surface of insect trypomastigotes, blood trypomastigotes and produced during human infection recognize the 83 kDa glycoprotein adhesion molecule by immunoblotting, indicating that this molecule that mediates this critical step is immunogenic and is a candidate for vaccination against Chagas' disease. PMID- 3046615 TI - A single amino acid substitution converts cytochrome P450(14DM) to an inactive form, cytochrome P450SG1: complete primary structures deduced from cloned DNAS. AB - Genes for lanosterol 14-demethylase, cytochrome P450(14DM), and a mutated inactive cytochrome P450SG1 were cloned from S. cerevisiae strains D587 and SG1, respectively. A single nucleotide change resulting in substitution of Asp for Gly 310 of cytochrome P450(14DM) was found to have occurred in cytochrome P450SG1. In this protein the 6th ligand to heme iron is a histidine residue instead of a water molecule, which may be the ligand for the active cytochrome P450(14DM). Molecular models of the active sites of the cytochrome P450(14DM) and cytochrome P450SG1 were built by computer modeling on the basis of the known structure of that of cytochrome P450CAM whose crystallographic data are available. The mechanisms which may cause a histidine residue to gain access to the heme iron are discussed. PMID- 3046617 TI - Biosynthesis of cholesterol in the yeast mutant erg6. AB - A mutant (erg6) of Saccharomyces cerevisiae defective in S-adenosylmethionine: delta 24-sterol-C-methyl transferase (EC2.1.1.41) which normally produces cholesta-5,7,24-trienol and cholesta-5,7,22,24-tetraenol as the major sterols (total 4,4-desmethyl sterol content-8.3 fg/cell) was shown to synthesize trace levels of cholesterol (0.08 fg/cell). The identity of cholesterol was established by co-chromatography in TLC, GLC and HPLC with an authentic sample, mass spectroscopy and after an incubation with [1-14C]acetate by isotopic dilution and recrystallization of the radiochemically purified material to constant specific activity. PMID- 3046619 TI - Trauma: an annotated bibliography of the recent literature. PMID- 3046618 TI - Peptide hormones and neuropeptides. Proteolytic processing of the precursor regulatory peptides. AB - Regulatory peptides are initially synthesized as large protein precursors which must undergo proteolytic processing to yield the biologically active peptide. The proteolytic enzymes involved in this post-translational processing remove the signal peptide, a hydrophobic peptide located at the amino-terminus of the precursor. Further processing steps are the cleavage at single or two adjacent basic residues by endoproteases, followed by a removal of basic residues located at the C-terminus of the peptides by a carboxypeptidase-like enzyme. The selective processing may play an important role in determining which peptides are finally released by the cell. This article reviews the selectivities of various enzymes known to play a role in the proteolytic processing of various regulatory peptides. PMID- 3046620 TI - Some effects of assisted reproduction on perinatal care. PMID- 3046616 TI - Nerve growth factor (NGF) is present in human placenta and semen, but undetectable in normal and Paget's disease blood: measurements with an anti-mouse NGF enzyme immunoassay using a recombinant human NGF reference. AB - An enzyme immunoassay (EIA) originally developed for mouse beta-nerve growth factor (NGF) and commercially available was validated for human NGF. Cell culture medium containing bioactive recombinant human NGF was used as a reference, and mouse 2.5S beta-NGF as a standard. One of three human placentas contained measurable NGF (70 pg/g of tissue of mouse beta-NGF equivalents), a second detectable, and a third undetectable NGF. In three human semen samples NGF content ranged from 0.13 - 1.4 ng/ml. NGF could not be detected in normal human serum and in plasma from patients with Paget's disease, although mouse 2.5S beta NGF added to human blood could be completely recovered from the serum. PMID- 3046621 TI - In vitro fertilization: reflections on the state of the art. PMID- 3046622 TI - The risk of high multiparity with IVF/ET. PMID- 3046623 TI - Perspectives on youth suicide. PMID- 3046625 TI - The role of the uteroplacental bed in pregnancy-related hypertension. PMID- 3046624 TI - The case for treating hypertension in the elderly. AB - Not so long ago it was fashionable to ignore a little bit of hypertension in an elderly person. The rationalizations for this position included the limited epidemiological data establishing the risk of elevated blood pressure in the age group over 65, particularly for women; and concern that lowering the blood pressure might not be desirable, from the standpoint of reduced blood flow to vital organs. There was also concern over the known greater responsiveness or sensitivity of older people to customary doses. It was also argued by some that even if the blood pressure could be safely controlled it was hardly worth it, since age itself is the most powerful predictor of mortality in the elderly. Moreover, it seemed unlikely to many that the risk of hypertension could be reduced--that is, that life expectancy could be extended; at the same time it seemed fairly certain that the quality of life would be compromised by therapy. However, since systolic blood pressure continues to be predictive of coronary heart disease mortality well into the ninth decade, and since considerable life expectancy remains for those who have survived to their senior years, it seems logical to attempt to reduce the risk and preserve life expectancy by controlling blood pressure. The question is, is the evidence that treatment helps really convincing, or are we simply extrapolating from studies carried out in younger subjects? Review of the early literature, which focused almost exclusively on stroke survivors and the influence of hypertension on stroke recurrence, provides inconsistent, conflicting, and not persuasive information.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3046626 TI - The combination of a low-Na/high-K salt with metoprolol in the treatment of mild moderate hypertension. A multicenter study. AB - To extend our previous findings that a low-Na/high-K salt (S) reduces BP in hospitalized patients, a multicenter study was performed. After a placebo period during which patients were informed by written instruction how to avoid only foods with a high Na content, 143 out-patients (84 males and 59 females, mean age 50.7 years, range 28-69) with DBP greater than or equal to 95 mm Hg randomly received for 4 weeks either metoprolol (M) 200 mg SR qd (67 patients), or S, 2 g bid to add to foods (76 patients). At the end of this period patients with DBP still greater than 90 mm Hg combined the two treatments for a further 4 weeks. Mean blood pressure (mm Hg), HR (bpm), 24-hrs urinary Na and K excretion were measured fortnightly. In comparison to pretreatment values MBP was significantly (P less than 0.01) reduced by both treatments, although to a greater extent in the M group already at the second week, without any further decrement thereafter. In the S group MBP decreased by 4.4 mm Hg and 27/76 patients were responders (DBP less than or equal to 90 mm Hg), while in the M group it was reduced by 9.0 mm Hg and 28/67 patients were responders. In the S group urinary Na excretion was significantly (P less than 0.01) lower than in the M group, and this difference was present until the end of period 1.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3046628 TI - Use of captopril to reduce serum lipids in hypertensive patients with hyperlipidemia. AB - The effects on serum lipids of substituting captopril for other antihypertensive drugs were evaluated in 24 essential hypertensives with stable hypercholesterolemia (total cholesterol 250 to 350 mg/dL). Captopril, 50 mg bid, was substituted for one of the nondiuretic drugs the patients were already taking. Patients on a diuretic continued the same diuretic at the same dosage. After 6 months of treatment including captopril, patients returned for 3 months to their previous therapeutic scheme (rechallenge). Treatment with captopril caused a reduction in total cholesterol (-18%) and triglycerides (-26%) and an increase in high-density lipoprotein (HDL) cholesterol (+27%). When patients returned to their precaptopril therapy, total and HDL cholesterol returned to precaptopril values. These effects of captopril on lipid profile of hypertensive patients with hypercholesterolemia could be important for the reduction of cardiovascular complications in treated hypertensives. PMID- 3046627 TI - Long-term captopril therapy with no effect on glucose metabolism in patients with essential hypertension. AB - The long-term antihypertensive response to captopril (25-50 mg/day) and 75g oral glucose tolerance (75g oGTT) following a 6-10 month period of captopril administration was evaluated in 20 patients with essential hypertension without persistent proteinuria. Eleven of these 20 patients exhibited impaired glucose tolerance (IGT), while the remaining nine patients had normal glucose tolerance (NGT). All patients tolerated long-term captopril therapy with no untoward effects. Six months' administration of captopril significantly decreased blood pressure in patients with NGT from 171 +/- 12/105 +/- 5 mm Hg (mean +/- SE) to 146 +/- 7/88 +/- 4 mm Hg. Also in patients with IGT, long-term captopril therapy decreased blood pressure from 165 +/- 3/97 +/- 2 to 143 +/- 5/86 +/- 2 mm Hg. No patient with NGT developed diabetes mellitus. Neither fasting nor post-glucose load venous blood glucose deteriorated in any of the patients during the therapy. There were no significant changes in the insulinogenic index (delta IRI/delta BG at 30 minutes after glucose load) in both the patients with NGT and IGT. In patients with IGT, the concentration of glycosylated hemoglobin (Hb A1 and Hb A1c slightly but significantly decreased from 8.1 +/- 0.3 to 7.7 +/- 0.4% (P less than 0.05) and from 5.9 +/- 0.3 to 5.5 +/- 0.3% (P less than 0.01) after 6.2 +/- 1.4 months' captopril therapy. These results suggest that in addition to its antihypertensive effects, long-term captopril therapy does not compromise glucose metabolism in hypertensive patients. PMID- 3046629 TI - Treating black hypertensives with capozide. AB - Twenty-four black men with mild to moderate essential hypertension were enrolled in an open-label trial comparing the efficacy of two doses of Capozide (captopril and hydrochlorothiazide). All antihypertensive drugs were discontinued and patients then received placebo for 2 weeks. Twenty-two patients, mean age 59.1 +/ 14.3 years, with sitting diastolic blood pressure (BP) 92 to 110 mm Hg, entered the 6-week active-drug phase. Eleven patients (Group A) were randomized to Capozide 25/15 and 11 (Group B) to Capozide 50/15. Baseline mean BPs were 151.0/100.7 mm Hg in Group A and 153.1/100.7 mm Hg in Group B. At week 6, mean BPs were 128.7/84.4 mm Hg in Group A and 126.8/82.7 mm Hg in Group B. Uric acid, blood urea nitrogen and creatinine levels rose slightly in both groups. There were no adverse events. Eighteen patients had normal BPs at study completion. Twice-daily Capozide treatment is effective and well tolerated in blacks; patients responded equally well to both doses. PMID- 3046630 TI - Treatment of mild hypertension with urapidil or captopril. AB - The aim of this study was to compare the efficacy (lowering diastolic blood pressure to 90 mm Hg or less) of urapidil, a postsynaptic alpha-blocker with central action with that of captopril in a randomized, double-blind, multicenter study. Two hundred ninety-five essential hypertensives (WHO I/II) (140 male, 155 female, age 56-60 years) were treated for 12 weeks with either 30 to 90 mg urapidil bid or 12.5 to 50 mg captopril bid. Supine blood pressure values (mmHg, mean +/- SD) at the end of the 12-week treatment dropped for the urapidil group (n = 142) from 175/103 +/- 19/6 to 154/89 +/- 17/9 (P less than 0.001), and in the captopril group (n = 153) from 175/103 +/- 19/6 to 154/90 +/- 19/9 (P less than 0.001), corresponding to 62% urapidil and 58% captopril responder rates. The results demonstrate that the two antihypertensive medications with different modes of action control blood pressure with equal efficacy at three dose levels. A dose decrease was possible in 20% of each group, but a dose increase was necessary in 39% of urapidil and 44% of captopril group. PMID- 3046631 TI - Multicenter comparison of dilevalol to placebo in patients with mild hypertension. AB - The antihypertensive effects of oral dilevalol, a beta-blocking agent with vasodialating properties were compared with placebo in 128 mildly hypertensive patients (supine diastolic blood pressure 95 to 105 mm Hg) in a multicenter, double-blind, parallel group study. Following a 4-week placebo phase, 63 patients were randomly assigned to receive dilevalol and 65 to receive placebo. A titration phase followed during which the dose of dilevalol was increased every other week from 100 mg to 800 mg to achieve a supine diastolic blood pressure (SDBP) less than 90 mm Hg and decreased by 10 mm Hg or more from baseline. A matching number of placebo capsules for each dose level of dilevalol was dispensed for blinding purposes. Patients who had at least a 5 mm Hg reduction in SDBP entered a 1-month maintenance phase. Blood pressure and heart rate were measured weekly 20 to 24 hours after a dose. This study demonstrated that the minimally effective dose of dilevalol was 100 mg, which was shown to result in a significantly (P less than or equal to 0.05) greater reduction in systolic BP. A once-daily dose of 200 mg was superior to placebo in both systolic and diastolic BP effects (P less than 0.001 and less than 0.001, respectively), and this superiority was seen again at both the end of titration and the end of the study. The BP reduction was accompanied by a small (5 bpm) decrease in heart rate. The side effect profile of dilevalol was not different from placebo. Dilevalol appears to have no adverse effect on plasma lipids.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3046632 TI - Taste acuity and zinc metabolism in captopril-treated hypertensive male patients. AB - Hypogeusia is a reported side effect of captopril. Linkage of hypogeusia to zinc deficiency has been suggested. We objectively assessed taste acuity using Henkin's three-drop stimulus technique and measured plasma zinc (PZn) level and urinary zinc excretion in 31 hypertensive patients. Of these, 11 were long-term, high-dose captopril recipients (more than 6 months, 266 +/- 34 mg/day), six were short-term captopril recipients (less than 6 months, 104 +/- 40 mg/daily dose), and the remaining 14 served as noncaptopril controls. Compared to controls, the long-term captopril group had significantly higher taste detection and recognition thresholds, lower PZn (91 +/- 3 vs. 100 +/- 3 micrograms/dl, P less than 0.05) and higher urinary zinc excretion (1017 +/- 89 vs. 609 +/- 76 micrograms/day, P less than 0.005). The short-term captopril group did not differ significantly from the noncaptopril group except for higher taste-recognition thresholds for NaCl and sucrose (P less than 0.05). Discontinuing captopril improved taste acuity and almost normalized zinc parameters in two patients on long-term captopril. These results suggest that abnormalities of taste are commonly associated with captopril therapy and may be related to changes in zinc metabolism. This is especially true in patients on long-term, high-dose captopril therapy. PMID- 3046634 TI - Serotonin, atherosclerosis, and collateral vessel spasm. AB - Studies on animal models demonstrate that platelet products contribute to vascular spasm in ischemic syndromes and that this is reversible with administration of ketanserin and thromboxane synthesis inhibitors. Laboratory animals (dogs, rabbits, and rats) that had femoral artery ligations exhibited supersensitivity to serotonin within days in their collateral blood vessels. This supersensitivity lasted at least 6 months. The response to serotonin was reversed by ketanserin, but not by 5HT-1 antagonists. Supersensitivity does not extend to norepinephrine, and alpha blockers do not influence the response to serotonin. It appears that platelet activation by endothelial injury contributes to ischemia through blood vessel occlusion and vascular spasm. When platelet activation occurs in vivo, blood vessel occlusion and vascular spasm are reversible in part by using ketanserin or agents that block thromboxane synthesis or its action. Combining both classes of agents reverses spasm completely. These findings support existing evidence that platelet products contribute to vascular disease, and provide an approach to improved management with currently available pharmacologic agents. PMID- 3046633 TI - Ketanserin in essential hypertension. Effects of blood pressure on renal hemodynamics. AB - Essential hypertension is a multifactorial disorder mediated by multiple mechanisms. Serotonin may cause vasoconstriction either directly or indirectly by amplifying the actions of vasoconstrictor substances. Clinical trials indicate that ketanserin, a serotonin antagonist, is effective in treating hypertension. It is desirable for an antihypertensive agent to enhance or preserve renal hemodynamic function. In this study, we evaluated the acute (1 week) and chronic (8 weeks) effects of ketanserin (20-40 mg twice a day) on glomerular filtration rate, renal plasma flow, and sodium excretion in patients with uncomplicated hypertension. Patients with untreated diastolic blood pressure (greater than 90 mm Hg) received either ketanserin or a placebo during an 8-week double-blind trial. Findings demonstrated that patients treated with ketanserin showed an increase in renal plasma flow compared with those who received placebo. Sodium excretion remained unchanged, indicating the absence of sodium retention during therapy. These results show that ketanserin lowers blood pressure in essential hypertension while preserving renal hemodynamics and function. PMID- 3046635 TI - Effects of a new serotonin antagonist, ketanserin, in experimental and clinical hypertension. AB - Ketanserin is an agent whose main pharmacologic action is antagonism of serotonin (5-hydroxytryptamine, 5HT) receptors of the 5HT2 subtype. It also has weak alpha 1-adrenergic blocking properties, which may contribute to the acute blood pressure lowering effects seen in animal models of hypertension. During chronic treatment of hypertension in animals, the 5HT2 antagonistic properties, or a combination of 5HT2 and alpha 1-antagonistic effects, seems to be responsible for ketanserin's hypotensive action. Studies of patients with hypertension have demonstrated the therapeutic effects of ketanserin in monotherapy and combination therapy. In humans, the drug has a terminal half-life of 12-25 hours, and a twice daily dosage will lower blood pressure over the day. Ketanserin is a vasodilator that acts on both resistance and capacitance vessels. Chronic treatment with the drug causes minimal reflex changes in cardiovascular function, as well as sustained blood pressure reduction comparable with the effects of beta-adrenergic blockers or diuretics. In elderly patients, the therapeutic effects of ketanserin appear to be greater, and side effects are less frequent compared with beta blockers and diuretics. In addition to its antihypertensive action, ketanserin also produces other effects that may be important in reducing cardiovascular morbidity and mortality in patients with hypertension. Some studies have shown a reduction in total and low-density lipoprotein (LDL) cholesterol, and a rise in high-density lipoprotein (HDL) cholesterol. Ketanserin also reduces ex vivo platelet aggregation and inhibits the serotonin-induced platelet release reaction. Worldwide experience with ketanserin in the treatment of hypertension indicates that it is a safe and effective agent for long-term therapy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3046636 TI - The absence of long-term therapeutic effect of calcium channel blockade in the primary aldosteronism of adrenal adenomas. AB - The absence of long-term effect of calcium channel blockers in primary aldosteronism due to adrenal adenoma is described in two patients. Calcium entry channel blockers, which were claimed to be a new medical therapy for primary aldosteronism, neither prevented the increase of plasma aldosterone nor decreased potassium levels over time in these patients. The drugs did not even lower blood pressure in one patient. Whereas calcium channel blockers may be effective in adrenal hyperplasia, they are apparently not effective enough in the long term treatment of patients with primary aldosteronism due to adrenal adenoma. PMID- 3046637 TI - Inflammatory mediators in chronic otitis media with effusion. AB - Otitis media with effusion (OME) is a common middle ear inflammatory disease in the pediatric population. This article determines concentrations of three functionally and metabolically distinct inflammatory mediators in middle ear effusions (MEE) and corresponding plasma of children with OME. One hundred two patients (mean age, 4.9 years) with persistent OME were studied. Middle ear effusions were collected from all subjects and plasma from a subset at the time of tympanostomy tube insertion. Histamine was assayed radioisotopically, 13,14 dihydro-15-keto-prostaglandin F2 alpha (stable PGF2 alpha metabolite) by radioimmunoassay, and neutrophil chemotactic factor of anaphylaxis by modified Boyden chamber. Mean MEE levels of the mediators (39 +/- 13 ng/mL, 462 +/- 179 pg/mL, and 264% +/- 57% positive control, respectively) were markedly higher than those of corresponding plasma (0.5 +/- 0.1 ng/mL, 285 +/- 127 pg/mL, and 47% +/- 5% positive control, respectively). The mean histamine content of mucoid effusions (43.2 +/- 56.9 ng/mL) was significantly higher than that of purulent (22.5 +/- 10.5 ng/mL) and serous (17.9 +/- 16.8 ng/mL) effusions. Higher histamine levels were observed in effusions positive for Haemophilus influenzae when compared with those with other pathogenic isolates. The high concentrations of these mediators in MEE and their potential for inducing or sustaining the inflammatory process supports a role in the pathogenesis of OME. PMID- 3046638 TI - Randomized surgical adjuvant trial of interferon alfa-n1 in recurrent papillomatosis. AB - Sixty-six patients with recurrent respiratory papillomatosis of juvenile onset were treated for six months with interferon alfa-n1 (Wellferon) in a randomized crossover trial. Half received interferon alfa-n1 intramuscularly at a dosage of 5 megaunits per square meter daily for 28 days and then thrice weekly for five months, followed by six months of observation. The other half were observed for six months and then treated. Operations were performed every two months to assess disease extent by a scale developed for this purpose. The score for the patients during the first observation period was stable. There was a statistically significant lowering of score in patients receiving interferon alfa-n1 during both periods of drug administration. Eight of 57 patients with assessable airway disease achieved complete remission, as did one additional patient with disease limited to the nasopharynx. No patients achieved complete remission during six months of observation alone. This difference was statistically significant. Patients without tracheostomy were significantly more likely to achieve remission than those with a tracheostomy. The patients who were observed after discontinuation of the drug therapy showed a significant rise in score within four months. Symptoms of toxicity included transient fever, fatigue, nausea, and headache. Elevations in serum aspartate aminotransferase levels occurred in 64% of the patients. There was an inverse correlation between age and the ability to tolerate the medication. The dose studied may be close to the maximum tolerated dose. It appears that interferon alfa-n1 as an adjuvant to routine surgical management is effective in slowing the growth of respiratory papillomas. PMID- 3046639 TI - Before the big time: early history of the Training School at Vineland, 1888 to 1949. AB - Leaders of the Training School at Vineland made major contributions to the field of mental retardation from 1890 to 1940. S. O. Garrison originated the cottage plan and emphasized education and self-support. E. R. Johnstone pioneered teacher training, published a journal, initiated social action programs, and established a research laboratory. H. H. Goddard standardized the Binet-Simon scales in the United States, validated heredity as a cause of mental retardation, and hosted the production of the first group intelligence tests. S. D. Porteus supplemented the Binet with measures of personality. E. A. Doll established that birth injury could cause mental retardation, investigated the educability of profoundly retarded individuals, produced the Vineland Social Maturity Scale, and fostered electroencephalographic research. PMID- 3046640 TI - Adherence of Campylobacter jejuni and Campylobacter coli to porcine intestinal brush border membranes. AB - The adhesion of Campylobacter jejuni and C. coli to isolated porcine intestinal brush border membranes was studied by phase-contrast and electron microscopy. Approximately 45% of the cell population adhered to the brush borders, possibly in a specific manner. Pretreatment of the brush borders with trypsin or pronase, and competitive inhibition with L-rhamnose caused a slight reduction of the adhesion. Different forms of pretreatment of the bacterial cells reduced their ability to adhere, but also their motility. PMID- 3046641 TI - Erythropoietin-induced secondary polycythemia in a patient with a renal cell carcinoma. A case report. AB - This study provides clear documentation of in vivo biogenesis of erythropoietin (Epo) by a human renal carcinoma. A middle-aged woman with a clear cell renal carcinoma of the left kidney developed severe polycythemia. This polycythemia was accompanied by markedly elevated levels of immunoreactive erythropoietin both in the peripheral venous blood, and in blood derived from the left renal vein during nephrectomy. Exstirpation of the non-invasive renal carcinoma was followed by complete restoration of both hematocrit and erythropoietin plasma concentration to normal levels. The fall in plasma erythropietin concentration immediately after nephrectomy (T/2 less than or equal to 3 hours) was probably a valid representation of plasma erythropoietin metabolism in this patient. Direct evidence of erythropoietin production in individual renal carcinoma cells was provided by immunoperoxidase studies demonstrating focal cytoplasmatic accumulation of immunoreactive erythropoietin in the tumor cells. PMID- 3046644 TI - The place of radiation therapy in the treatment of squamous cell carcinoma of the nasal vestibule. A review. AB - Radiation therapy and limited resection give equally good results in early squamous cell carcinomas of the nasal vestibule, each producing local control rates of 90% or more. For more extensive disease, primary radiation therapy with surgery reserved for residual or recurrent carcinoma is recommended in view of the significant cosmetic defects which generally follow major resection. Small and superficial lesions can be treated by external beam therapy or interstitial implants. Large or infiltrative lesions are best managed by external beam therapy. Serious late morbidity following irradiation is uncommon and has been reported in fewer than 5% of patients. Regional nodal metastases are diagnosed at the time of first presentation in about 5% of patients and signal a very poor prognosis. However, the data available do not support elective treatment of clinically uninvolved regional nodes. Fewer than 5% of patients manifest late nodal metastases when the primary tumor area remains free of recurrence, and most of these metastases can be controlled by neck dissection and/or irradiation. PMID- 3046642 TI - Extracellular deposit of the cationic proteins ECP and EPX in tissue infiltrations of eosinophils related to tissue damage. AB - In a series of eosinophil inflammatory states affecting various organs (heart, gut, bladder and skin) we performed an immunohistochemical study of the eosinophil cationic proteins ECP and EPX. A strong correlation was noted between the liberation of ECP and EPX and tissue necrosis in all organs. In most cases ECP and EPX were found on the same location. However, one case indicated a possible differential release. Extracellular ECP and EPX were revealed concurrently with the two polyclonal antibodies and the monoclonal EG2 antibody. The latter binds to both ECP and EPX, but only during secretion. Since EG2 does not differentiate between ECP and EPX, but only during secretion. Since EG2 does not differentiate between EXP and EPX, it is for the first time demonstrated that both cationic proteins are correlated to tissue damage. The chymotrypsin-like cationic protein (CCP), related to neutrophils, showed a low correlation with the eosinophil cationic proteins in cases of tissue damage. The hypothesis is put forward that the release of eosinophil granule proteins and especially ECP results in a non-specific tissue damage. PMID- 3046643 TI - Cancer cure with organ preservation using radiation therapy. AB - In 1987, the American Cancer Society has anticipated that 965,000 new cases of invasive cancer will be diagnosed in the United States. About 40% of those patients can be treated for cure with organ preservation using radiation therapy. The basic biologic background for such an approach to the problem has become well established. The clinical data to substantiate the validity of the concept date from 1902 until the present. Organ preservation has become a major and important concept in the management of the patient with invasive cancer. New and innovative techniques for treatment are enabling the organ to be preserved, the cancer to be cured, and appropriate cosmesis and function to be preserved. Many tumor sites are appropriate for this treatment technique, including breast, eye, larynx, prostate, soft tissue sarcomas, etc. PMID- 3046645 TI - Long term clinical outcome of coronary surgery and assessment of the benefit obtained with postoperative aspirin and dipyridamole. AB - Three hundred and twenty patients originally entered into a randomised study to assess the effect of aspirin and dipyridamole on the patency of coronary bypass grafts one year after operation were clinically reassessed a mean of 6.6 years (range 4.3-8.6) after operation. Patients were recruited between 1978 and 1982 after the present policy of total revascularisation had been adopted. During the follow up period there were 25 deaths of which 17 were due to cardiac causes (average annual cardiac mortality 0.8%). Of 280 patients available for contact, 250 (89.3%) attended an outpatient interview. Ninety four (37.6%) patients complained of recurrent angina but in only 23 (9.2%) was this severe. Two hundred and eleven (84.4%) of the 250 patients underwent exercise stress testing. There were 73 (34.6%) abnormal tests of which 52 were in the group of 94 patients with recurrent angina. Myocardial infarction occurred in nine of the 250 patients during the follow up period. Twenty six patients (10.4%) had reinvestigation for symptoms. This group had a graft occlusion rate of 52%. Half these patients have required reoperation and 20 of 22 occluded or severely stenosed grafts were replaced. In only two instances were vein grafts inserted into vessels with new disease. Half of the original group were given aspirin (330 mg three times a day) plus dipyridamole (75 mg three times a day). Of the 250 patients interviewed, 122 took aspirin and dipyridamole from the second postoperative day for a mean of 25 months, with warfarin for three months. The other 128 patients took placebo for a mean of 23 months together with warfarin for three months. This long term treatment with aspirin plus dipyridamole conferred no significant benefit for all clinical outcomes measured at a mean of 6.6 years. PMID- 3046648 TI - To practice my profession faithfully. PMID- 3046647 TI - Ischaemic ventricular aneurysms: true or false? PMID- 3046646 TI - Comparison of the haemodynamic effects of epoprostenol (prostacyclin) and tolazoline. AB - The haemodynamic effects of infusion of epoprostenol (prostacyclin) and bolus injection of tolazoline were compared in a crossover study in 11 children with pulmonary hypertension caused by pulmonary vascular disease. The children were studied during cardiac catheterisation, while they were anaesthetised, paralysed, and ventilated with 100% oxygen. The order of drug administration was not randomised because tolazoline has a half life of hours whereas epoprostenol has a half life of a few minutes. Both drugs caused pulmonary and systemic vasodilatation, and there were no significant differences between the two. The 95% confidence intervals suggest that tolazoline did not have a clinically important haemodynamic advantage over epoprostenol. Previous reports suggest that serious side effects are common when tolazoline is used in repeated doses; epoprostenol has only a few minor side effects that are rapidly reversible when the infusion is stopped. Epoprostenol is more expensive than tolazoline but this study suggests that epoprostenol is a more suitable pulmonary vasodilator if more than a single dose is required. PMID- 3046649 TI - Low level exposure to asbestos: is there a cancer risk? PMID- 3046651 TI - A double-blind randomized controlled trial on the use of prophylactic antibiotics in patients undergoing elective caesarean section. AB - In a double-blind randomized controlled study 232 patients undergoing elective lower segment caesarean section were randomly allocated to receive a pre operative prophylactic dose of a combination of crystalline penicillin and chloramphenicol or a placebo. The two groups were comparable in terms of patient characteristics and operation variables. The group receiving antibiotics had significantly fewer febrile and infectious morbid events and thus spent fewer days in hospital than the group receiving the placebo. PMID- 3046652 TI - Uteroplacental and umbilical artery blood velocity waveforms in placental abruption assessed by Doppler ultrasound. Case report. PMID- 3046653 TI - Commentary: the safety of diagnostic ultrasound. PMID- 3046650 TI - Did Princess Charlotte die of pulmonary embolism? AB - The cause of death of the Princess Charlotte of Wales in 1817 has been attributed to postpartum haemorrhage. Although an autopsy was performed, pulmonary embolism was not diagnosed because this condition was not recognized until 1846, but the clinical description of her last hours clearly points to pulmonary embolization as the most likely cause of death. PMID- 3046654 TI - Control of astigmatism in cataract surgery. AB - A study is reported on cataract surgery, with intraocular lens implant, with measurement of the preoperative astigmatism and of the postoperative astigmatism over 28 weeks. Nine interrupted 10/0 nylon sutures are used to close a limbal section. Preoperative astigmatism is compensated for in the method of suturing by the placement of additional sutures. Postoperatively sutures are cut in line with the plus cylinder axis in eyes showing excessive astigmatism with the rule. Final postoperative astigmatism is controlled within 2.25 D cyl. 68% of cases lie within 1.0 D cyl with the rule to 1.0 D cyl against the rule. The average case in which sutures are not cut is one having 1.51 D cyl with the rule at one week postoperatively, declining to zero at approximately 12 weeks, and having a final value of 0.17 D cyl against the rule. No significant change in cylinder is seen after 10 weeks. The final postoperative astigmatism is only weakly correlated with the preoperative astigmatism, showing that the surgical method is effective. The spherical equivalent error is shown to shift in the direction of myopia in the postoperative period. PMID- 3046655 TI - Severe herpetic keratitis. I: Prevalence of visual impairment in a clinic population. AB - We report a prevalence study of the best visual acuity in the affected eye of 100 selected patients with herpetic keratitis seen during a two-year period. Sixty two patients retained an acuity of 6/9 or better without requiring penetrating keratoplasty (PK). The prevalence of reduced visual acuity severe enough to warrant PK was 33%. Patients requiring PK for whom full clinical records were available suffered a mean of 6.8 episodes of keratitis. In this group of patients the vision of 18 fell from 6/6 to 6/60 over a mean period of 8.5 years. Once visual acuity was permanently reduced to 6/12, 78% of patients proceeded to lose vision to 6/60. Unilateral visual impairment occurs in at least a third of patients with severe herpetic keratitis. Once vision falls permanently to 6/12, the long-term prognosis for vision appears to be poor. PMID- 3046656 TI - Acquired Brown's syndrome in a patient with combined lichen sclerosus et atrophicus and morphoea. AB - A 49-year-old woman with generalised lichen sclerosus et atrophicus and morphoea developed bilateral Brown's syndrome. Some of the skin lesions were in the vicinity of the trochlea. A characteristic feature of morphoea is subcutaneous fibrosis, so we postulate that the cause of the Brown's syndrome was mechanical tethering of the superior oblique tendon by deep subdermal fibrosis. Histopathological diagnosis was made from biopsies of similar lesions on the patient's face. PMID- 3046658 TI - Choroidal osteoma presenting in pregnancy. AB - A case of choroidal osteoma presenting during pregnancy is reported. The clinical findings are summarised and the features of choroidal osteoma are discussed. PMID- 3046657 TI - Effect of single-dose ivermectin therapy on human Onchocerca volvulus infection with onchocercal ocular involvement. AB - Ivermectin has shown promise as a potentially safe and effective microfilaricidal drug for the treatment of onchocerciasis. Several limited studies have shown it to have fewer side effects, especially ocular complications, than the currently available drug, diethylcarbamazine. The detailed ocular findings in 200 moderately to heavily infected Liberians who were enrolled in a safety and dose finding study are presented. They received either 0, 100, 150, or 200 micrograms/kg of ivermectin and were followed up for 12 months. In clinical studies so far carried out ivermectin in a dose of 100, 150, or 200 micrograms/kg has not been associated with any major adverse reactions nor were there any sight threatening effects even in the presence of severe ocular disease. Each of these doses significantly reduced the ocular microfilaria load for at least 12 months when compared with either the placebo (p less than 0.05) or pretreatment values (p less than 0.001). However, the 100 and 150 micrograms/kg doses caused fewer minor side effects than the higher dose. These results confirm that ivermectin in a single oral dose may be a safe and effective microfilaricidal drug for the treatment of onchocerciasis and that it appears to be free of major ocular side effects. PMID- 3046660 TI - Radicals in biological catalysis. PMID- 3046659 TI - Infectious crystalline keratopathy. AB - We present a patient who developed a crystalline keratopathy after a penetrating keratoplasty. This rare complication can be caused by bacterial infection, and the patient responded to the appropriate antibiotic therapy. The literature is reviewed and possible causes and mechanisms of the crystalline appearance are discussed. PMID- 3046662 TI - [3H]-p-azidopuromycin photoaffinity labeling of Escherichia coli ribosomes: evidence for site-specific interaction at U-2504 and G-2502 in domain V of 23S ribosomal RNA. AB - Previously we (1) showed that [3H]-p-azidopuromycin was a functional puromycin analogue that, on photolysis in the presence of 70S ribosomes from Escherichia coli, photoincorporated site specifically into proteins L23, L18/22, and L15 [Nicholson, A.W., Hall, C.C., Strycharz, W.A., & Cooperman, B.S. (1982) Biochemistry 21, 3809-3817] and (2) used immunoelectron microscopy to localize the principal sites of p-azidopuromycin photoincorporation within the 50S subunit [Olson, H.M., Nicholson, A.W., Cooperman, B.S., & Glitz, D.G. (1985) J. Biol. Chem. 260, 10326-10331]. These studies are here continued by identification of the principal sites of [3H]-p-azidopuromycin photoincorporation into ribosomal RNA. The major portion of such photoincorporation, 72%, takes place into 23S rRNA. Analysis by hybridization of the photoaffinity-labeled rRNA to restriction enzyme fragments of plasmid pKK3535, which contains rrnB DNA, using a refinement of a recently developed methodology [Hall, C.C., Smith, J.E., & Cooperman, B.S. (1985) Biochemistry 24, 5702-5711], shows that the most prominent [3H]-p azidopuromycin photoincorporation occurs within bases 2445-2668 in domain V [Noller, H.F. (1984) Annu. Rev. Biochem. 53, 119-162] of 23S rRNA. Photoincorporation into this region is site specific, as demonstrated by the decrease in photoincorporation of radioactivity when unlabeled puromycin is included in the photolysis solution. Significant site-specific photoincorporation also occurs within bases 489-681 in domain II of 23S RNA. Further localization, by the method of reverse transcriptase primer extension [Barta, A., Steiner, G., Brosius, J., Noller, H.F., & Kuechler, E. (1984) Proc. Natl. Acad. Sci. U.S.A. 81, 3607-3611], provides evidence that U-2504 and G-2502 are the principal sites of p-azidopuromycin interaction.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3046663 TI - Peptides mimicking the flap of human renin: synthesis, conformation, and antibody recognition. AB - Four peptides related to human renin flap region have been synthesized. Two of them are ring closed through appropriately designed disulfide bridges. Structure analysis involving IR and NMR techniques and recognition by polyclonal human renin antibodies provides support for a beta-hairpin secondary structure of the cyclized peptides identical with that presented by the flap section in the speculative human renin model [Blundell, T., Sibanda, B. L., & Pearl, L. (1983) Nature (London) 304, 273-275; Sibanda, B. L., Blundell, T., Hobart, P. M., Fogliano, M., Bindra, J. S., Dominy, B. W., & Chirgwin, J. M. (1984) FEBS Lett. 174, 102-111]. PMID- 3046664 TI - Site-directed mutageneses of rat liver cytochrome P-450d: axial ligand and heme incorporation. AB - By oligonucleotide-directed mutageneses, 13 substitutions of amino acids at the carboxy-terminal region of rat liver cytochrome P-450d were done as follows: (A) Phe-449----Tyr; (B) Gly-450----Ser; (C) Leu-451----Ser; (D) Gly-452----Glu; (E) Lys-453----Glu; (F) Arg-454----Leu; (G) Arg-455----Gly; (H) Cys-456----Tyr; (I) Cys-456----His; (J) Ile-457----Ser; (K) Gly-458----Glu; (L) Glu-459----Ala; (M) Ile-460----Ser. The CO-bound reduced forms of the wild type and mutants B-G, J, L, and M gave Soret peaks at 448 nm. The CO complex of mutant A gave a Soret peak at 445 nm. The intensities of the CO-bound forms of mutants A, C, D, and J were very small compared with that of the wild-type complex. The CO-reduced forms of mutants H, I, and K did not give a Soret peak around 450 nm at all. The 448-nm peak of mutant F was unstable and quickly disappeared with the concomitant appearance of a peak at 420 nm.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3046661 TI - Physical properties of DNA in vivo as probed by the length dependence of the lac operator looping process. AB - Plasmid constructs containing a wild-type (O+) lac operator upstream of an operator-constitutive (Oc) lac control element exhibit a length-dependent, oscillatory pattern of repression of expression of the regulated gene as interoperator spacing is varied from 115 to 177 base pairs (bp). Both the length dependence and the periodicity of repression are consistent with a thermodynamic model involving a stable looped complex in which bidentate lac repressor interacts simultaneously with both O+ and Oc operators. The oscillatory pattern of repression with distance occurs with a period approximating the helical repeat of DNA and presumably reflects the necessity for proper alignment of interacting operators along the helical face of the DNA. In the length regime examined, the presence of the upstream operator enhances repression between 6-fold and 50-fold depending upon phasing. This reflects a torsional rigidity of DNA in vivo that is consistent with in vitro measurements. The oscillatory pattern of repression is best fit with a period of either 9.0 or 11.7 bp/cycle but not 10.5 bp/cycle. This periodicity is interpreted as reflecting the average helical repeat of the 40-bp interoperator region of plasmid DNA in vivo, suggesting that the local helical repeat of DNA in vivo may differ significantly from 10.5 bp/turn. The apparent persistence length needed to fit the data (aapp) is only one-fifth the standard in vitro value. This low value of aapp may be due in part to DNA bending induced by catabolite activator protein (CAP) bound to its site between the interacting operators.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3046665 TI - Properties of the transferrin associated with rat intestinal mucosa. AB - The transferrin that is isolated from washed intestinal mucosal cell preparations consists partly of a fraction that has properties distinguishing it from serum transferrin. The serum transferrin contaminating mucosal preparations, even when fully saturated with iron and in the presence of proteinase inhibitors, also acquires the properties of the mucosal transferrin when the mucosa is homogenised. The mucosal transferrin is modified by a single cleavage of the polypeptide chain yielding a disulphide-linked peptide of 6550 daltons linked to the parent protein by a disulphide bridge. The amino-terminal sequence of the first 11 residues of this peptide could be aligned with both the known rat and human transferrin carboxy-terminal sequences. In both cases the sequence is preceded by a phenylalanine residue (residue 622 of human transferrin). This suggested that a mucosal chymotryptic enzyme was responsible even though rat transferrin is not susceptible to alpha-chymotrypsin if fully iron-saturated. Since transferrin mRNA is not found in the intestinal mucosa it must be imported from the serum. It remains uncertain whether the modified transferrin is present naturally and plays a role in iron absorption but these findings do indicate the eventual fate of any transferrin imported into an intestinal cell. PMID- 3046666 TI - Major malformations of the urinary tract. Anatomic and genetic aspects. AB - This paper outlines the problem of major malformations of the urinary tract, their frequency and their etiology. It is based on data analysis of 900 neonatal autopsies performed in our center from 1978 to 1987. These major malformations are often lethal and they can essentially be separated into disorders of the kidney and malformations of the lower urinary tract. Single-gene disorders are often seen in the former whereas the second group is more likely to be multifactorially controlled. We do insist on the importance of a strict collaboration between pathologists and geneticists. PMID- 3046667 TI - Renal response to vasopressin in premature infants: what is new? AB - Clinical and experimental data indicate that the neonatal pituitary is capable of responding with arginine vasopressin (AVP) release to physiological stimulation and pathological events commonly seen in the perinatal period. The limited concentrating performance of the newborn kidney is thought, therefore, to be accounted for by the diminished end-organ responsiveness to AVP and the inability of the immature kidney to generate and maintain deep corticopapillary osmotic gradient. Evidence has been provided to indicate that in low birth weight premature infants the impaired renal tubular sodium transport, the renal salt wasting and late hyponatremia, and the increased renal prostaglandin production may be important factors in attenuating renal response to AVP. PMID- 3046669 TI - Renal function and fluid therapy in high risk infants. AB - In recent years, the survival rate of high risk infants has markedly increased. The role of such medical management as fluid, electrolyte and nutritional therapy have assumed a greater importance in assuring optimal quality of the survivors. The very low birth weight infants, particularly those with respiratory distress syndrome and perinatal asphyxia, are at highest risk. The inefficient renal function, unique characteristic of body fluid composition and/or presence of severe clinical illness often make the management of fluid and electrolytes in this group of infants difficult. The numerous factors that influence insensible water loss make calculation of fluid management in the high risk infant even more challenging. Systematic collection of data such as daily body weight, intake, output, urine specific gravity and serum electrolyte is essential to appropriately maintain fluid and electrolytes balance in these infants. Respiratory distress syndrome is a common problem in premature infants and the fluid and electrolyte management in these infants will require similar attention to details as described for the fluid and electrolytes of very low birth infants. Perinatal asphyxia often results in oliguria or anuria because of possible development of inappropriate ADH secretion or acute tubular necrosis. It is essential that fluid restriction be done on the first day or two of life to avoid fluid overload. PMID- 3046668 TI - Renal aspects of calcium and phosphorus metabolism in preterm infants. AB - The aim of this study is to emphasize renal aspects of calcium and phosphorus metabolism from our data of more than 200 metabolic balance studies carried out in preterm infants. Renal production of 1,25-dihydroxyvitamin D increased rapidly after birth provided the concentration of the substrate, 25-hydroxyvitamin D, is adequate. The gut of preterm infants is able to respond to the active metabolite of vitamin D. Mean plasma phosphate threshold for tubular reabsorption of phosphate is high, about 2.1 mmol/l or 6.5 mg/dl. The low fractional excretion of phosphate cannot be explained by immature parathyroid function nor by renal unresponsiveness to parathormone, at least after the first days of life. It is probably due to regulating factors related to the high rate of growth. Because of reduced glomerular filtration rate, a too high phosphorus intake may result in hyperphosphatemia. Conversely, a too low phosphorus intake will lead to a phosphate depletion syndrome characterized by marked increase in urinary calcium excretion, no urinary phosphate, and hypophosphatemia. Preterm infants with chronic metabolic acidosis are able to acidify urine so that titratable acid is directly related to urinary excretion of phosphate. Clinical implications are that calcium:phosphorus ratio in milk must be adapted according to net bone and soft tissue retention. PMID- 3046670 TI - Functional renal insufficiency: role of adenosine. AB - Adenosine, a direct degradative product of 5'AMP, could be a physiological regulator of glomerular filtration rate and renal blood flow, acting on the renal arteriolar tone and the tubuloglomerular feedback mechanism. Recent experimental evidence strongly suggests an important role for intrarenal adenosine in the hemodynamic renal changes observed in postocclusive acute renal failure, in hypoxemia-induced renal insufficiency, as well as in various experimental models of acute renal failure. In these conditions, adenosine appears to induce predominant postglomerular vasodilatation, as well as preglomerular vasoconstriction when the renin-angiotensin system is activated. Theophylline, a xanthine with strong adenosine antagonistic properties, prevents the hypoxemia induced renal changes, while enprofylline, a xanthine devoid of such properties, does not prevent these changes. Available data favor the hypothesis that intrarenal adenosine is involved in the pathophysiology of the renal changes observed in the early phase of vasomotor nephropathy. PMID- 3046671 TI - Aminoglycoside nephrotoxicity and urinary excretion of N-acetyl-beta-D glucosaminidase. AB - The purpose of this paper is to discuss briefly the mechanism of aminoglycosides nephrotoxicity. This kind of antibiotic seems to act preferentially on the phospholipid metabolism of the proximal tubular cell. A lysosomal enzyme, N acetyl-beta-D-glucosaminidase, could be of interest in assessing this renal interference. PMID- 3046672 TI - Low-frequency collective motion in biomacromolecules and its biological functions. AB - The occurrence of low-frequency motion in biomacromolecules, which had long been speculated upon with a great deal of skepticism, is now a clearly established phenomenon and has been convincingly demonstrated. The next stage in the process of its elucidation appropriately concerns the determination of its origin, the development of a feasible and effective model for the calculations involved and, more importantly, the extending of investigations on its biological roles in order to gain insights into the various interesting mechanisms underlying the dynamic processes occurring in biomacromolecules. Confronted with such a task, this review has been written with the aim of stimulating further, through a systematic and comprehensive description, developments to open up this exciting frontier of molecular biology, especially from the viewpoint of biological functions. PMID- 3046673 TI - [Effectiveness of methods of isolating specific class-G rabbit antibodies for the immunoenzyme determination of human placental alkaline phosphatase]. AB - Four different chromatographic methods of IgG isolation from rabbit antisera to placental alkaline phosphatase (HPAP) have been compared. The antibodies were obtained by ion-exchange chromatography and affinity chromatography on protein-A sepharose, on the sepharose with immobilized antigen. IgG samples were characterized by the content of specific antibodies to HPAP and checked in enzyme immunoassay (EIA). IgG purified on immobilized antigen were found to be the optimal both from the point of view of the specific antibodies content and EIA sensitivity, but satisfactory results could be also obtained with ion-exchange and protein-A-chromatography purified IgG. The last two isolation methods are simpler and provide 3-10 ng/ml sensitivity of HPAP detection, which is lower, as compared with the test employing affinity antibodies (1 ng/ml), but allows the detection of HPAP in serum samples. PMID- 3046674 TI - [Differences in the excitatory action of acetylcholine and mecholine on mechanosensitive high-threshold C-axonal units of the skin in the cat]. AB - Intraarterial acetylcholine (Ach) and acetyl-beta-methylcholine (metacholine, Mch) excite feline high-threshold C-fiber mechanosensitive cutaneous sensory units (SU) in a different way. Ach in noxious (algogenic for humans) concentrations of 10 micrograms/ml and more induces a high-frequency discharge, with mean frequency of impulses at its peak being 4-12 Hz. This discharge seems to result from activation of N-cholinoreceptors because non-noxious and nonalgogenic M-agonist Mch as well as Ach in subnoxious concentrations induce in SU only a low-frequency discharge with a mean frequency of impulses 0.5-3.5 Hz. The data are discussed in connection with nociception and tissue chemoreception. PMID- 3046675 TI - [ICO-13 monoclonal antibodies to the differentiation antigen of human hemopoietic cells]. AB - Monoclonal antibodies (Mab) ICO-13 of IgM isotype reacted in indirect surface immunofluorescence test with 88.5 +/- 3.4% of thymocytes and 7.1 +/- 2.4% of T cells, but failed to react with other peripheral blood cells from healthy donors. The antigen disappeared in cells stored at 4 degrees C overnight or at -196 degrees C in liquid nitrogen. The antigen detected by Mab ICO-13 was expressed on blast cells of acute lymphoblast leukemias and lymphomas. The antigen was absent in chronic lymphoblast leukemia and blast crisis of chronic myeloid leukemia. PMID- 3046676 TI - [Immunohistochemical study of the carcinoembryonic antigen (CEA) in human tumors using the CEA-specific oncoprecipitin crustacin]. AB - 58 human tumors have been studied immunohistologically with the aid of CEA specific precipitin--crustacin (Cr). The results were in general similar to those which were achieved with anti-CEA-antibodies. But there were some differences. The reaction with Cr in the cytoplasm was mostly diffuse, while the reaction with Ab was localized usually in the peripheral parts of the cytoplasm. The reaction with Cr has a more restricted character and is more expressed in ovarian tumors and less expressed in gastric and colonic tumors than the reaction with Ab. PMID- 3046677 TI - [The role of host cells in the genetic regulation of plasmid transfer in Escherichia coli]. AB - The role of serologically typed and nontyped E. coli cells in the expression of genetic regulation systems activity for repressed plasmids transfer (types fin OP, fin U, fin V) was studied. It is shown that the inhibiting action of such plasmids on the tra-genes functions of derepressed plasmids is more expressed in the cells of serologically nontyped E. coli (K-12 strains) than in the cells of serologically typed ones (serogroups O106, O128, O147). PMID- 3046678 TI - [Effect of fibroblast growth factor and nerve growth factor on the cellular proliferation and functional activity of isolated islands of Langerhans]. AB - The effects of fibroblast growth factor (FGF) and nerve growth factor (NGF) on DNA synthesis and insulin secretion were studied in 4-5-day cultures of the isolated neonatal rat islets. FGF (0.1 ng/ml) stimulated significantly the incorporation of 3H-thymidine into DNA of the isolated islets, but failed to change either insulin content in the islets or the rate of insulin secretion. NGF (0.1-1000 ng/ml) did not affect the above parameters. The responses of the islets of Langerhans to increasing concentrations of glucose and isobutylmethylxanthine were not modified after prolonged exposure to NGF. The role of FGF and NGF in the regulation of proliferation and secretory process in pancreatic islet cells is discussed. PMID- 3046679 TI - [Morphological evaluation of reconstructive operations on the bronchi using a microsurgical technic in an experiment]. AB - Morphological study of bronchial anastomoses repair made with or without microsurgical techniques has been performed in 30 mongrel dogs. Comparative analysis of bronchoscopy, microscopy and scanning electronic microscopy data shows earlier epithelialization, more regular conjunction of bronchial tube and less marked inflammation in anastomosis made with microsurgical techniques. The fact may be explained by a less pronounced traumatization of bronchial tissues with microinstruments and also by tight suture of mucosal layer which creates favourable conditions for repair of anastomosis per primam. PMID- 3046682 TI - A simplified bone marrow cryopreservation method. PMID- 3046680 TI - Clinicopathologic, immunophenotypic, and immunogenotypic analyses of immunoblastic lymphadenopathy-like T-cell lymphoma. AB - Immunoblastic lymphadenopathy (IBL)-like T-cell lymphoma is a distinct peripheral T-cell lymphoma, which closely resembles angioimmunoblastic lymphadenopathy with dysproteinemia (AILD) and/or IBL, but is characterized by focal or sheet-like proliferation of immunoblasts and pale cells of T-cell nature. In this report, 36 patients with IBL-like T-cell lymphoma were analyzed. The disease is clinically characterized by generalized lymph node swelling, hepatosplenomegaly, fever, skin rash, polyclonal hypergammaglobulinemia, marked male predominance, predilection for the elderly, and poor prognosis. There was no association with human T-cell leukemia virus type I or human immunodeficiency virus. IBL-like T-cell lymphoma may be divided into two categories (CD4+ type and CD8+ type) by surface marker analysis. It can also be divided into three categories on the basis of the histologic findings of distribution of morphologically recognizable tumor cells: nine cases of "inconspicuous type," six cases of "patchy type," and 21 cases of "diffuse type." Two cases of "inconspicuous type" converted later to "diffuse type." DNA hybridization analyses in the ten recent cases revealed that three of four "inconspicuous types" and five of six "diffuse types" showed clonal rearrangement of T-cell receptor-beta chain gene without rearrangement of immunoglobulin heavy chain gene, providing strong evidence for clonal proliferation of T cells. PMID- 3046681 TI - Donor buffy coat cell infusion after marrow transplantation for aplastic anemia. PMID- 3046684 TI - Quantitation and identification of human monocytic colony-stimulating factor in human serum by enzyme-linked immunosorbent assay. AB - An enzyme-linked immunosorbent assay (ELISA) system for the quantitation of human monocytic colony-stimulating factor (hM-CSF) was established, which was based on the "dual antibody immunometric sandwich" principle using horse and rabbit polyvalent antibodies against human urinary colony-stimulating factor (CSF-HU). The minimal detectable level of hM-CSF was 10 U/mL, and the assays showed good reproducibility. As measured by this method, the average serum hM-CSF level of 20 normal adults was 540 +/- 110 U/mL (range, 300 to 800 U/mL). The peak of hM-CSF measured by ELISA was identical to that measured by bioassay when semipurified CSF-HU was fractionated by reversed-phase high performance liquid chromatography (HPLC). This method detected two types of hM-CSF, which had approximate molecular weights of 85 Kd (CSF-HU) and 45 Kd in human serum and urine; the ratio of 85:45 Kd was very high in serum and the amounts of the two types were nearly equal in urine. After anticancer chemotherapy, the serum hM-CSF level of one half of the patients with hematological malignancy was elevated according to the reduction in neutrophil number, while it was almost in the normal range in the other half of the patients, indicating the possibility that anticancer chemotherapy damaged the hM-CSF-producing cells. This ELISA method may be useful for monitoring the serum hM-CSF level after anticancer chemotherapy. PMID- 3046683 TI - Prominent expansion of circulating lymphocytes bearing gamma T-cell receptors, with preferential expression of variable gamma genes after allogeneic bone marrow transplantation. AB - The presence of two distinct T-cell receptors (TCR) alpha/beta dimers or gamma/delta dimers, was systematically analyzed in peripheral blood lymphocytes form 26 recipients of allogeneic bone marrow transplants for leukemia. When using monoclonal antibody WT31, which recognizes a common epitope on the alpha/beta heterodimer, the expansion of peripheral CD3+, WT31- cells to 40% of the PBLSs was detected in two patients. In patient 2, the presence of circulating TCR gamma bearing cells was directly demonstrated with monoclonal antibody Ti gamma A directed against the V gamma 9 J gamma p gene products. From CD3, WT31- clones derived from patients 1 and 2, sequential immunoprecipitations were performed with anti-CD3 and anti-C gamma to determine the CD3-associated structure. Molecular weights of gamma subunits were different in both patients, thus indicating structural heterogeneity. The ability of TCR gamma clones to proliferate when stimulated with anti-CD3 beads was observed for clones from patient 2, whereas this response required exogenous interleukin-2 for clones from patient 1. We have already shown that the TCR gamma cells from patient 1 might have played a role in the immunodeficient state. Similar conclusions cannot be drawn from patient 2. Southern blot analysis of total PBL gamma cell lines and clones indicated that this major circulating subset of TCR gamma cells retained a TCR beta gene in germline configuration and preferentially expressed a single V gamma gene, V gamma 5 for patient 1 and V gamma 9 for patient 2. PMID- 3046685 TI - Stimulation of neutrophil production in CSF-1-responsive clones. AB - The hematopoietic growth factor CSF-1 has been considered relatively lineage specific for the production of macrophages, whereas GM-CSF elicits a predominance of neutrophils. It is likely that in vivo, individual clones are stimulated by the two CSFs, although the effect of dual stimulation on progenitors and their progeny has not been completely explored. We found that in cultures initiated with low concentrations of CSF-1 or GM-CSF, alone or in combination, production of macrophages predominated. Maximally stimulatory concentrations of CSF-1 elicited a predominance of macrophages, whereas maximal GM-CSF elicited many more neutrophil/macrophage colonies and pure neutrophil colonies. A combination of maximal CSF-1 and GM-CSF elicited the same differentiation as GM-CSF alone. Delayed addition of GM-CSF to cultures initiated with CSF-1 elicited colonies indistinguishable from GM-CSF alone, suggesting that neutrophil production had been switched on by GM-CSF. In mapping studies, colonies initiated by CSF-1 increased or switched on neutrophil production when GM-CSF was added as a second stimulus. These studies show that individual clones are responsive to both CSFs, and that the differentiating influence of GM-CSF predominates over that of CSF-1. In cultures to which only CSF-1 was added, a population of progenitors was sustained that produced neutrophils only after a GM-CSF stimulus. Thus, CSF-1 may participate in maintaining a reserve of progenitors for neutrophils during periods of increased neutrophil demand. PMID- 3046686 TI - Therapeutic potential of recombinant granulocyte-macrophage colony-stimulating factor and interleukin-3 in murine B-cell leukemia. AB - The antileukemic activity of murine recombinant granulocyte-macrophage colony stimulating factor (rGM-CSF) and a combination of rGM-CSF and recombinant interleukin-3 (rIL-3) was examined by using a murine model of spontaneous B-cell leukemia (BCL1) in BALB/c mice. All untreated mice inoculated with 2 x 10(2) BCL1 cells developed leukemia within 4 weeks, with extreme lymphocytosis and a massive increase in both spleen weight and cell number while the number of myeloid progenitors (CFU-C) per spleen was decreased. In contrast, rGM-CSF-or rGM-CSF- and rIL-3-treated recipients did not show any evidence of leukemia or splenomegaly at 4 weeks and showed a significant increase in CFU-C per spleen. Hematologic parameters in the peripheral blood of untreated mice showed anemia and thrombocytopenia. Significant elevations in these parameters were recorded in mice treated with either protocol of CSF. Treatment of recipient mice with either rGM-CSF or rGM-CSF and rIL-3 prolonged their median survival from 6 weeks in untreated controls (range, 5 to 9 weeks) up to the time they were killed at 105 days. Adoptive transfer of spleen cells obtained from mice treated with rGM-CSF, mice treated with a combination of rGM-CSF and rIL-3, and untreated controls, into normal secondary recipients indicated improved survival in recipients inoculated with rGM-CSF. These data indicate that CSFs may inhibit in vivo expansion of leukemic cells of lymphoid origin. PMID- 3046687 TI - Relapse cell population differs from acute onset clone as shown by absence of the initially activated N-ras oncogene in a patient with acute myelomonocytic leukemia. AB - We have conducted a follow-up study of a patient with myelomonocytic leukemia exhibiting an N-ras mutation (Gln61----Lys61) using the polymerase chain reaction method and synthetic oligonucleotide hybridization probes. This method allowed us to detect as little as 3% of N-ras-mutated cells within a population. When the patient went into clinical remission, the mutation became undetectable. When a relapse occurred, the blasts did not carry the N-ras mutation. Analysis of M13 cloned amplified N-ras sequences from relapse DNA revealed exclusively the wild type allele of the N-ras gene. These findings suggest that the relapse cell population is derived from a different clone than the acute phase population. Furthermore, the data argue that N-ras mutation is not an initiating lesion in this case of acute myelomonocytic leukemia (AMML). PMID- 3046688 TI - Cytogenetics of malignant lymphomas. PMID- 3046689 TI - Correlation between low CSA plasma concentration and severity of acute GvHD in bone marrow transplantation. AB - Between 1982 and 1986 51 patients were treated with ciclosporin a (CSA) to prevent graft versus host disease (GvHD) after bone marrow transplantation (BMT). Major side effects of the drug were tremor, hypertension, hepatotoxicity and nephrotoxicity. Acute GvHD 0 degree to II degree occurred in 80% of our patients, and GvHD III degree and IV degree in 20% despite the use of CSA. Two to four days before the onset of GvHD, CSA serum levels were significantly lower on the average in patients who developed GvHD III degree and IV degree compared to the others. Our data indicate that plasma CSA concentrations higher than 250 ng/ml should be achieved to reduce the severity of GvHD after BMT. PMID- 3046690 TI - Comments on Hodgkin's disease: the Sternberg-Reed cell by P. Bucsky. PMID- 3046691 TI - [Recent data on the cytogenetics of colorectal adenocarcinoma]. AB - A brief description of the results obtained by cytogenetic analyses of cancer cells from colorectal adenocarcinomas is reported. Two distinct patterns of chromosomal anomalies are observed. The major one, called "monosomic type", because many chromosomes losses exist, is characterised by the losses or deletions of the following chromosomes 18, 17 (short arm = p), 1p, 4, 14, 5 (long arm = q) and 21. It frequently evolves towards polyploidy, by duplication of all remaining chromosomes. The minor one, called "trisomic type", is characterised by the gain of several chromosomes: 7, 12, X, 5 and 8. The chromosomal anomalies observed seem to have no topological relationships with oncogenes, and other interpretations for their occurrence were investigated. The numerous and frequent deletions of some chromosomes, like 17p and 18, may indicate the involvement of recessive genes, like in the anti-oncogene system. In addition, it is observed that most deletions involve genes for "de novo" pathways whereas gains involve genes for salvage pathways of the synthesis of nucleotides. A correlated cytogenetic and enzymologic study was thus developed, and the first results show that the chromosomal pattern may well indicate the metabolic deviations. Monosomic type tumors have, on the average, a relatively low "de novo" and a high salvage pathways, and trisomic type tumors a high "de novo" and salvage pathways. Since chemotherapy is principally orientated against "de novo" pathways of the synthesis of nucleotides, these results may help to understand the difficulties of chemotherapy in colorectal cancers and perhaps to adapt it better. PMID- 3046692 TI - [Prognostic and therapeutic impact of the cellular morphology and type of bone marrow invasiveness in multiple myeloma]. PMID- 3046693 TI - [French Society of Quantitative Microscopy. New methods of microscopy and analyses and cytometry; pathologic anatomy: contribution to diagnosis, case finding and research; automated analysis of the genome. Paris, 15-16 March 1988. Abstracts]. PMID- 3046694 TI - A Virtual Notebook for biomedical work groups. AB - During the past several years, Baylor College of Medicine has made a substantial commitment to the use of information technology in support of its corporate and academic programs. The concept of an Integrated Academic Information Management System (IAIMS) has proved central in our planning, and the IAIMS activities that we have undertaken with funding from the National Library of Medicine have proved to be important extensions of our technology development. Here we describe our Virtual Notebook system, a conceptual and technologic framework for task coordination and information management in biomedical work groups. When fully developed and deployed, the Virtual Notebook will improve the functioning of basic and clinical research groups in the college, and it currently serves as a model for the longer-term development of our entire information management environment. PMID- 3046695 TI - Digital subtraction dynamic cavernosography. AB - Digital subtraction angiography (DSA) was used in the investigation, by dynamic cavernosography, of 35 men with acquired erectile impotence. The benefits of using DSA included shorter examination times, lower contrast medium dosage and better definition of abnormal veins. In particular deep crural veins are more clearly seen than in conventional studies and these may be of more importance than was previously thought. PMID- 3046697 TI - Variations in surgical practice: welcome diversity of disturbing differences. AB - The review examines the variations in surgical practice and assesses the value of surgical audit in quality control and allocation of surgical resources. PMID- 3046696 TI - The appearances on ultrasound of the female urethral sphincter. AB - A rounded or ovoid midline structure with mean measurements of 1.30 cm x 1.33 cm x 0.96 cm in longitudinal, transverse and antero-posterior dimensions was routinely imaged at the bladder base in 97 female patients on pelvic ultrasound examination. Its position and appearance are reminiscent of a smaller version of the male prostate, and it has been dubbed the "female pseudoprostate". It appears to correspond with the external rhabdo-sphincter of the bladder. Its rounded shape may be confusing but it should not be misread as pathological. PMID- 3046698 TI - Traumatic retroperitoneal venous haemorrhage. AB - Vena caval injury is rare in the UK, but changes in society suggest that it may become more frequent. The problem is always unexpected and presents a major surgical challenge. Modern techniques have led to a substantial improvement in survival so that most patients with an infrarenal injury should now survive but only with prompt, effective treatment. In this review the problems and techniques of surgical management are illustrated by three cases of traumatic injury and two of peroperative accidents. PMID- 3046699 TI - Management of major colonic haemorrhage. AB - Major colonic haemorrhage remains a difficult diagnostic and therapeutic problem. We propose that those patients who continue to bleed after resuscitation are best served by immediate laparotomy. High flow antegrade irrigation and intra operative colonoscopy can then be used to localize the site of bleeding and allow appropriate excisional surgery rather than blind colonic resection. PMID- 3046700 TI - The task of a statistical referee. AB - The results are presented of an audit of the statistical standard of papers published in The British Journal of Surgery. A number of deficiencies are highlighted, many of which stem from an over-emphasis on statistical significance at the expense of any assessment of the clinical relevance of research findings. The flaws in the design of published studies, and in particular the many instances of inadequate sample sizes, emphasize that statistical input should be sought at the beginning of a research project rather than at the end. PMID- 3046701 TI - Intravenous digital subtraction angiography and Duplex scanning in the detection of late human umbilical vein degeneration. AB - To define the exact incidence of late degeneration, 32 patients underwent intravenous digital subtraction angiography (IVDSA) and/or a Duplex scan more than 3 years after human umbilical vein (HUV) grafting. IVDSA (n = 26) showed a 23 per cent aneurysmal degeneration rate which increased to 40 per cent with Duplex scanning (n = 25). Although the differences obtained in those patients receiving both examinations (n = 19) were statistically not significant, Duplex scanning appeared to be more sensitive, demonstrating two additional cases of aneurysmal degeneration not detected by IVDSA. Since, moreover, Duplex scanning proved able to detect anastomotic stenosis, it appears to be the examination of choice in long-term follow-up of the HUV graft. Despite this relatively high degeneration rate, the authors consider continued use of the HUV graft in selected patients to be justified, especially when the long-term patency rates, the available alternatives, and the less favourable long-term survival after femoropopliteal reconstruction are taken into consideration. PMID- 3046703 TI - Changing nature of anal cancer. PMID- 3046702 TI - Treating the premenstrual syndrome. PMID- 3046705 TI - ABC of eyes. Acute visual disturbance. PMID- 3046704 TI - Idiopathic retroperitoneal fibrosis. PMID- 3046706 TI - Ultrasonography for diagnosing appendicitis. PMID- 3046707 TI - ABC of eyes. History and examination. PMID- 3046709 TI - Central organization of wave localization in the clawed frog, Xenopus laevis. II. Midbrain topology for wave directions. AB - The organization of water wave localization within the midbrain of the clawed frog Xenopus was investigated by performing behavioral tests on frogs that had partial midbrain ablations. The criterion of localization was the orientation of turns toward the origin of impinging waves. All lesion effects became apparent as localization failure within an angular sector of wave direction. The sectors were contralateral to the lesion and of various sizes, some comprising the complete hemifield. Localization outside of the sectors was not affected. Thus, wave localization is topologically organized with respect to wave direction. Two topological projections were found. Lesions of the first projection resulted in unoriented responses to the affected wave directions. After lesions of the other projection, the frog responded to waves from the affected directions by lunges without turns. It is suggested that the two types of localization failures are due to impairment of the sensory wave direction detection and of the sensorimotor transfer, respectively. The essential midbrain areas are presumably the magnocellular nucleus of the torus magnocellularis and the ventrolateral tectum, but a considerable part of the localization might be done in the medulla. PMID- 3046708 TI - Central organization of wave localization in the clawed frog, Xenopus laevis. I. Involvement and bilateral organization of the midbrain. AB - The central nervous organization of water wave localization in the clawed frog Xenopus laevis was investigated by performing behavioral tests on frogs that had various brain ablations. The criterion of localization was the orientation of response turns toward the origin of stimulus waves. After complete midbrain ablation, Xenopus still detected impinging waves but could not localize them. After thalamopretectal ablation, however, Xenopus localized waves with normal accuracy. Thus, wave localization can be accomplished in the brainstem, and the midbrain is necessary for it. After forebrain ablation, the frogs no longer responded to water waves, which shows that higher brain centers modulate localization. Tectal lesions that spared the ventrolateral tectum did not abolish localization. After unilateral extirpation of tectum and torus, all ipsilateral waves were localized, but contralateral waves were not. This indicates a functional chiasm for the determination of wave directions in the midbrain. Total localization failure after unilateral midbrain destruction demonstrates that wave localization also requires the ipsilateral motorial tegmentum. When wave localization was abolished, a residual correlation between stimulus directions and response angles remained. PMID- 3046710 TI - Development of the retinofugal pathway in birds and mammals: evidence for a common 'timetable'. AB - We have compared changes in axon numbers in the developing optic nerves of eight homeotherms (seven mammals and one bird) using data from the available literature and our own material. The proportion of axons lost during development is smaller in the chick (35%) than in mammals (54-74%). The relative magnitude of this loss does not correlate with the extent of the binocular visual field or the size of the retinofugal ipsilateral projection. The timing of developmental events in the retinofugal pathway was compared as a proportion of the period between conception and eye opening (percentage of the 'caecal period', CP). In eutherian mammals, retinal ganglion cells and their target neurons are generated between 30 and 49% of CP (a duration of 19% of CP). In homeotherms, a phase of rapid axon generation begins around 38% of CP and the peak number of axons is reached at about 56% of CP (a duration of 18% of CP). A phase of rapid axon loss begins thereafter, and in most species it ends at about 74% of CP (a duration of 18% of CP), the rate of rapid axons loss being about half the rate of rapid axon generation. The similarity in relative timing (homochrony), like the similarity in the relative magnitude of the axon loss, suggests that cell generation and loss in the retinofugal pathway are influenced by a mechanism common to all homeotherms. We propose that in homeotherms each cohort of retinal ganglion cells is numerically matched with a group of target cells that is at an appropriate stage of maturation ('temporal matching' hypothesis). About twice as many ganglion cells are produced in each cohort than are needed, and their survival is determined by natural selection. PMID- 3046711 TI - Distribution, morphology and habenular projections of adenosine deaminase containing neurons in the septal area of rat. AB - Adenosine deaminase (ADA) was localized within several types of neurons within the septum and in septal efferent projections to the habenula by immunohistochemical, biochemical, retrograde tracing and lesion methods. Numerous ADA-immunoreactive (ADA-IR) neurons were observed in the septofimbrial nucleus, the triangular septal nucleus and the bed nucleus of the anterior commissure, while considerably fewer numbers were seen in the lateral septal area. Based on their size, shape and dendritic features, 4 morphologically distinct types of ADA IR neurons were recognized in these septal structures. In addition, fine, non varicose, ADA-IR fibers appeared to emanate from the postcommissural cell groups and these coalesced within the stria medullaris, continued caudally within this fiber bundle, and gave rise to a dense field of very fine immunoreactive elements within a restricted zone of the dorsal half of the medial habenula. Comparisons of the habenular localization of ADA-IR and enkephalin-IR elements showed that fibers labelled for either ADA or enkephalin occupied distinct, non-overlapping regions within the dorsal half of the medial habenula. After injections of Fluoro gold (FG) into the medial habenula, the majority of ADA-IR neurons in the septofimbrial nucleus, triangular septal nucleus, and the bed nucleus of the anterior commissure were retrogradely labelled with this fluorescent tracer, whereas no ADA-positive FG-labelled neurons were observed in the lateral septal region. Unilateral transections of the stria medullaris caused substantial depletions of ADA-immunoreactivity and reduced enzymatically determined ADA activity by up to 80% in the medial habenula on the lesioned compared with the contralateral control side. These results demonstrate that ADA-IR neurons in the septum are heterogeneously distributed and that populations of positive neurons within the postcommissural septal nuclei give rise to dense, focal projections to the medial habenula. These projections appear to be restricted to a portion of the medial habenula known to contain substance P-IR neurons and are subregionally segregated from enkephalin-positive septohabenular projections ending within this same portion. In addition to pointing out a unique capacity for adenosine catabolism within some septal neurons, possibly related to purinergic neuromodulation, the results indicate the utility of ADA-immunohistochemistry for the delineation of anatomical relationships between the septum and the medial habenula. PMID- 3046712 TI - A subpopulation of dopaminergic neurons in rat ventral mesencephalon contains both neurotensin and cholecystokinin. AB - The coexistence of the neuropeptides neurotensin and cholecystokinin and the catecholamine-synthesizing enzyme tyrosine hydroxylase within neurons of the ventral mesencephalon was analyzed using an immunofluorescence triple-labeling technique. Virtually all of the neurotensin-positive cell bodies in the ventral tegmental area, medial substantia nigra pars compacta, retrorubral field, and rostral and caudal linear raphe nuclei were found to contain both cholecystokinin and tyrosine hydroxylase immunoreactivities. The degree of colocalization was lower and more variable in other regions including the ventral and central periaqueductal grey matter and dorsal raphe nucleus. It appeared that immunoreactivities for these 3 neuroactive substances were not contained within the same axonal-like fibers and terminals in the ventral midbrain. These results demonstrate that a subpopulation of dopaminergic neurons, which presumably comprise part of the ascending mesotelencephalic system, contains the two peptides neurotensin and cholecystokinin. Thus, the data suggest a morphological basis for some of the reported functional interactions of these 3 putative neurotransmitters/neuromodulators within this system. PMID- 3046713 TI - An improved secondary structure computation method and its application to intervening sequence in the human alpha-like globin mRNA precursors. AB - Current secondary structure prediction computations have a serious drawback. The calculated thermodynamically most stable structure often differs from that observed in solution or in crystal form. In this paper we suggest a way to partially over-come some of these limitations by simulating the RNA folding process and calculating the frequencies of occurrence of the various substructures obtained. The frequently recurring substructures are then selected to construct the secondary structure of the whole RNA. 142 tRNA molecules and an E. coli 16S rRNA molecule have been examined by this method. The percentage of successful prediction of the correct helices are significantly higher than those calculated previously. The secondary structures of intervening sequences (IVSs) excised from human alpha-like globin pre-mRNAs are also computed. Thus, in this method the secondary structures obtained are composed of the statistically more significant substructures. This has also been demonstrated by using randomly shuffled sequences. The secondary structures of each of the randomized sequences are computed and their mean and standard deviations are used in evaluating the significance of the substructures obtained in the folding of the biological sequence. Some potentially appealing structural features aligning adjacent exons for ligation have been found. PMID- 3046714 TI - Discriminant analysis of promoter regions in Escherichia coli sequences. AB - We have previously developed a general method based on the statistical technique of discriminant analysis to predict splice junctions in eukaryotic mRNA sequences [Nakata, K., Kanehisa, M. and DeLisi, C. (1985) Nucleic Acids Res., 13, 5327 5340]. In order to evaluate further applicability of this method, we now analyze the promoter region of Escherichia coli sequences. The attributes used for discrimination include the accuracy of consensus sequence patterns measured by the perceptron algorithm, the thermal stability map, the base composition and the Calladine-Dickerson rules for helical twist angle, roll angle, torsion angle and propeller twist angle. When applied to selected E. coli sequences in the GenBank database, the method correctly identifies 75% of the true promoter regions. PMID- 3046715 TI - Treatment of periorbital giant nevi. PMID- 3046717 TI - [Oligodendroglioma of spinal cord]. PMID- 3046718 TI - The 1987 James A. F. Stevenson memorial lecture. The development of fetal behavioural patterns. AB - The development of fetal behaviour is reviewed. Fetal cutaneous and muscle sensory receptors are developed by the time movements are first seen. Human infants certainly respond to painful stimuli at 28 weeks. There is no clear evidence that prenatal "stress," e.g., maternal exposure to random noise and bright lights, impairs fetal development in the rat, on the contrary. Fetal diurnal rhythms appear in man and sheep before the development of sleep states; they are dependent on maternal corticosteroids, but the fetal mechanism is uncertain. With the development of sleep states (in late gestation in man and sheep, postnatally in the rat), the complex central control of behaviour is gradually established, but wakefulness is still of low incidence. The location of the sleep cycle generator is uncertain; the results of experimental lesions of the brainstem in fetal lambs appear incompatible with studies in adult rats and cats. PMID- 3046719 TI - Diabetes-induced rat tracheal segment supersensitivity to carbachol. AB - Contractile responses to carbachol of tracheal segments isolated (i) from rats made diabetic 4 months prior by a single intravenous injection (50 mg/kg) of streptozotocin (group B), and (ii) from diabetic rats that had been treated during the same period with a daily dose (2-4 U/animal) of long-acting insulin (group C) were compared with the contractile responses of trachea isolated from age-matched control animals (group A). Tracheal segments from group B were significantly more responsive to carbachol than those from group A or C at low, but not at high carbachol concentrations. Carbachol pD2 values were higher in group B (6.85 +/- 0.05) than in groups A (6.46 +/- 0.07) or C (6.37 +/- 0.06), but were not significantly different between groups A and C. These data indicate that diabetes induces a supersensitivity to carbachol in airway smooth muscles, possibly related to a diabetes-induced vagal autonomic neuropathy. PMID- 3046716 TI - Neonatal meningitis due to Proteus mirabilis: report of 3 cases. PMID- 3046720 TI - Effect of nickel chloride on streptozotocin-induced diabetes in rats. AB - The potential of nickel chloride to prevent streptozotocin-induced hyperglycemia was tested in rats in vivo. To induce diabetes, streptozotocin (100 mg/kg body weight) was injected as a single dose. Streptozotocin treatment resulted in a significant decrease in plasma insulin and ceruloplasmin, and pancreatic Cu, protein, and Cu-Zn superoxide dismutase activity. In rats treated with nickel chloride (10 mg/kg body weight) and streptozotocin, these values were comparable with those observed in control rats. The results indicate that nickel chloride injected before streptozotocin prevented streptozotocin-induced hyperglycemia, and suggest that the protective effect was related to Cu-Zn superoxide dismutase activity, mediated by copper. PMID- 3046721 TI - Interaction of Candida albicans with genital mucosa: effect of sex hormones on adherence of yeasts in vitro. AB - Findings from our previous studies revealed a correlation between the level of adherence in vitro of Candida albicans to human exfoliated vaginal epithelial cells (VEC) and the hormonal status of the cell donors. In the present study we investigated the effect of the sex hormones estradiol, estriol, progesterone, and testosterone on the binding of the yeasts to HeLa cell lines and VEC in vitro. Monolayers of HeLa cells were exposed to the hormones and yeasts under controlled conditions. The number of adherent yeasts per square millimetre of HeLa cell monolayers and the percentage of VEC with adherent yeasts was estimated by microscopic counts. The results showed that the tested sex hormones affected at various degrees the adhesion of yeasts to HeLa cells or VEC. Progesterone had the most marked effect, leading to a significant increase in the number of adherent yeasts to HeLa cells or in the percentage of adhesion of VEC. In addition, VEC were separated on Percoll gradients into the two cell types: superficial (S) and intermediate (I), cell types which appear physiologically under increased serum levels of estradiol or progesterone, respectively. Adhesion assays with the separated cell populations revealed an increased binding capacity of the I cells. The finding that progesterone increased the adherence of yeasts to genital mucosa and that VEC of the I type have a higher capacity to adhere the yeasts is compatible with our previous observation that increased numbers of I cells, appearing under high level of progesterone, are found in situations known to have predisposition to vaginal candidiasis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3046722 TI - The role of bacterial hydrophobicity in infection: bacterial adhesion and phagocytic ingestion. AB - The role that bacterial surface hydrophobicity (surface tension) plays in determining the extent of adhesion of polymer substrates and phagocytic ingestion is reviewed. The early attachment phase in bacterial adhesion is shown to depend critically on the relative surface tensions of the three interacting phases; i.e., bacteria, substrate, and suspending liquid surface tension. When suspended in a liquid with a high surface tension such as Hanks balanced salt solution, the most hydrophobic bacteria adhere to all surfaces to the greatest extent. When the liquid surface tension (gamma LV) is larger than the bacterial surface tension (gamma BV), then for any single bacterial species the extent of adhesion decreases with increasing substrate surface tension (gamma SV). When gamma LV less than gamma BV then adhesion increases with increasing gamma SV. Bacterial surface tension also determines in part the extent of phagocytic ingestion and the degree to which antibodies specifically adsorb onto the bacterium resulting in opsonization. The nonspecific adsorption of antibodies results in a considerable modification in the surface properties of the bacteria. Bacterial surface hydrophobicity can be altered significantly through exposure to subinhibitory concentrations of antibiotics, surfactants, lectins, etc. The effect of these changes on subsequent phagocytic ingestion is discussed. PMID- 3046723 TI - The human antibody response to uropathogenic Escherichia coli: a review. AB - Urinary tract infections caused by Escherichia coli are associated with a local and systemic antibody response. We have studied the serum and urine antibody responses to Escherichia coli in men and women with pyelonephritis, cystitis, and asymptomatic bacteriuria. Protein immunoblots consistently demonstrated serum antibody response to lipopolysaccharide (LPS). Anti-LPS antibody titres rose significantly and progressively when comparing acute with convalescent sera in those who have had their first urinary infection. For those with repeated infections, high titre LPS antibodies were present and did not change significantly between acute and convalescent sera. Antibody responses to the major outer membrane proteins were present but did not differ significantly when compared with normal human serum. A specific anti-P pilus antibody response was demonstrated by immunoblotting. Anti-P pilus antibody was quantitated using ELISA and the titres were found to be very low. Three other techniques were also used to demonstrate the presence of serum antibody. Antibody was detectable by immunofluorescence, but the antigenic specificity of the antibody was more difficult to ascertain. Immunoprecipitation was more specific for determining the nature of the antibody response. Lastly, immunoelectron microscopy was valuable in demonstrating antipilus and antiflagellar antibodies. Immunoelectron microscopy and immunoblotting provided evidence that human antiserum to P pili was modestly cross-reactive and could bind heterologous P pili. These studies indicated that the major antibody response in humans occurs after pyelonephritis and is directed against LPS. An anti-P pilus response is frequently present and is cross-reactive to some extent with other P pili. PMID- 3046724 TI - Immunogenetic aspects of host immune response. AB - The central role of histocompatibility leukocyte antigens (HLA) class II molecules in antigen presentation has received great attention in recent years, yet class I molecules have been defined as primarily functioning as a restriction element for cytotoxic T cell killing of virus-infected cells. Extensive clinical evidence, however, indicates that the HLA class I genes are strongly associated with nonseptic complications of enteric and genitourinary bacterial infections. Ninety percent of patients with Reiter's syndrome and reactive arthritis are positive for HLA-B27, yet the mechanism of disease susceptibility conferred by this gene remains obscure. Hypotheses concerning this interaction include (i) class I antigens functioning as receptors for microbial antigens; (ii) class I antigens expressing determinants that cross-react with microbial antigens; and (iii) class I genes controlling immunoregulatory functions that dictate qualitative differences in immune response to pathogenic organisms. These hypotheses await formal testing and hold great promise for understanding immunogenetic control of immune responses in general. PMID- 3046725 TI - The use of bacterial interference to prevent infection. AB - For decades, bacterial strains of low virulence were occasionally used in man to replace or to block colonization by the more virulent organisms and thereby prevent bacterial infection. This paper reviews the topic and presents recent information on the implantation of strain 215 alpha-hemolytic streptococcus (alpha-strep) in the nasopharynx of neonates in the intensive care unit. A single inoculation of strain 215 can change abnormal colonization of the pharynx to "normal" (alpha-strep predominant) in 48-72 h in most neonates. Following implantation, alpha-strep with strain 215 like characteristics fluctuate among naturally occurring strains of alpha-strep, sometimes persisting in dominance and sometimes decreasing rapidly as new strains appear. Strain 215 can survive in the pharynx during subsequent antibiotic therapy and can be recalled to dominance by such therapy. It seems remarkably stable in vivo. There is no evidence of its nosocomial spread in the nursery. Streptococcus with strain 215 like characteristics occurred naturally in 1-6% of neonates in our intensive care unit. No infection (disease) attributable to strain 215 occurred in implanted infants. PMID- 3046726 TI - The spectrum of Escherichia coli--Bacteroides fragilis pathogenic synergy in an intraabdominal infection model. AB - Pathogenic synergy between Escherichia coli and Bacteroides fragilis was investigated in an intraabdominal infection model. Defined inocula of E. coli and B. fragilis, alone or in combination, were enmeshed within a fibrin clot and surgically implanted into the peritoneal cavity of rats. A spectrum of bacterial synergy ranging from synergistic abscess formation to synergistic lethality was demonstrated using this model. The type of synergy exhibited was dependent upon the initial E. coli inoculum. When combined with B. fragilis, high inocula of E. coli (greater than 10(8) cfu/clot) produced synergistic lethality while low inocula (2 x 10(2) to 2 x 10(7) cfu/clot) resulted in synergistic abscess formation. With respect to abscess formation, there was reciprocal synergy between E. coli and B. fragilis. Abscesses resulting from mixed inocula were larger and had significantly higher numbers of E. coli and B. fragilis than abscesses initiated by monomicrobial inocula. These studies define a clinically relevant model of bacterial interactions in the setting of intraabdominal infection and suggest that conclusions drawn from experimental models of bacterial synergy should consider the type of model examined, the strains of bacteria studied, and the number of bacteria inoculated. PMID- 3046727 TI - Wounds of the abdomen. Part 2: In peace. AB - Until the mid 1960s in the United States, penetrating abdominal wounds in civilians were badly handled and often fatal. Little attention was paid to the methods used and results reported by military surgeons in previous wars such as those in Korea and Vietnam. In 1966 the National Academy of Sciences, National Research Council Committee on Shock pointed out that there was an increasing incidence of severe civilian injuries associated with unacceptably high morbidity and mortality and that this was because of lack of organized civilian community care. As a result the Trauma/Emergency Medical Service (T/EMS) was formed. The initiators based the organization on the remarkable success of military concepts and techniques. The improvement in the evacuation and treatment of penetrating abdominal wounds in peace from the inception of the T/EMS to the present is described in this paper. The methods used and results obtained through the T/EMS have made an important contribution to both civilian and military surgery--a unique event in peacetime. PMID- 3046728 TI - Mead Johnson Critical Care Symposium for the Practising Surgeon. 2. Complications of monitoring systems. AB - Current invasive monitoring techniques, although valuable in the care of critically ill patients, also have disadvantages. Complications of these techniques include errors in interpretation of measurements and complications of venous access and indwelling lines. To prevent these, the author recommends a standard routine for inserting and changing catheters, regular calibration of transducers and insertion of Swan-Ganz catheters with the balloon inflated with 1.0 to 1.5 ml of air. PMID- 3046729 TI - Mead Johnson Critical Care Symposium for the Practising Surgeon. 3. Monitoring and investigation of intra-abdominal sepsis. AB - Diagnosis and management of intra-abdominal sepsis continue to be major problems in critically ill patients. Multiple system organ failure secondary to intra abdominal sepsis continues to cause serious morbidity and death. The first step in management is to recognize the infection, while providing careful supportive therapy. A number of radiologic investigations, including ultrasonography and computed tomography, will help to diagnose a potential source of infection, which can be positively identified by fine-needle aspiration and culture. The septic focus must be drained either percutaneously or, if this fails, surgically. Use of specific antibiotics is imperative. Delay in diagnosis and surgery increases the death rate, so all available diagnostic modalities should be utilized, but these should not replace careful ongoing clinical assessment. PMID- 3046730 TI - Mead Johnson Critical Care Symposium for the Practising Surgeon. 4. Abdominal crisis in the intensive care unit. AB - Abdominal crises are common in critically ill patients who are admitted to the intensive care unit for problems unrelated to the abdomen. General surgeons may be asked to assess these patients for such reasons as pain, distension, possible sepsis, radiologic or laboratory abnormalities. Since many of the diagnostic signs and symptoms of acute abdomen are blunted or absent in critically ill patients who may be comatose or have been given analgesics or steroids, frequent thorough physical examination and close cooperation with the service admitting the patient are necessary to ensure early diagnosis and aggressive treatment of the abdominal crisis. PMID- 3046732 TI - Review of the surgical management of recurrent hiatal hernia: 5-year follow-up. AB - Symptoms in patients with hiatal hernia often respond to treatment consisting of diet and medication. Operative procedures, designed to control gastroesophageal reflux and avoid surgically induced problems, are reserved for those with intractable symptoms. When these operative procedures fail, reoperation may be necessary. The reoperative procedure is often technically complex because of esophageal and gastric scar fixation. The authors reviewed the surgical management of recurrent hiatal hernia in 168 patients followed up to 5 years or more; 43 of them had undergone gastric surgery previously.Radiologically, 97% patients studied (142 of 146) had no evidence of anatomic recurrence or reflux post operatively. Manometric studies postoperatively in 114 patients showed that the mean tone of the high pressure zone was within the normal range and lower esophageal disordered motor activity was decreased by 34.5% from the preoperative level. Symptoms of recurrent hiatal hernia were abolished by operation in 88% of the patients; only 4.8% had serious or recurrent symptoms. PMID- 3046731 TI - Mead Johnson Critical Care Symposium for the Practising Surgeon. 5. The diagnosis and management of common soft-tissue infections. AB - Soft-tissue bacterial infections range from the superficial, relatively benign form, readily treated by oral administration of antibiotics, to the highly lethal variety which requires extensive surgical debridement, administration of broad spectrum antibiotics and patient monitoring in the intensive care unit. The key to patient survival in the latter group is early diagnosis and treatment. Those at risk of serious infections include diabetic and cirrhotic patients and trauma victims who have been treated either surgically or medically. In addition to awareness of the possibility of serious infection, x-ray films of the soft tissue to show gas, aspiration of fluid collections for Gram's staining and local incision of the tissue to inspect fascia and muscle are all useful aids in diagnosis. This review attempts to classify soft-tissue infections and to recommend an approach to diagnosis and management. PMID- 3046733 TI - Liver blood flow after major hepatic resection. AB - The factors involved in liver regeneration are poorly understood, but it has been suggested that blood flow plays a role. This paper documents the changes in liver blood flow (LBF) that occur after major hepatic resection. Eight patients, ranging in age from 37 to 76 years, underwent liver resection. Liver blood flow was measured preoperatively and on days 1, 4 and 7 postoperatively by low-dose galactose clearance. There was a significant (p less than 0.01) fall in LBF on day 1 compared with the baseline value, followed by a significant (p less than 0.01) rise from the baseline value by day 4. By day 7, LBF had returned to baseline levels and was significantly (p less than 0.01) lower than on day 4. These changes in LBF may be related to the stimulus for liver regeneration and increased functional demands during the early regenerative phase. PMID- 3046734 TI - Extrication, immobilization and radiologic investigation of patients with cervical spine injuries. AB - Most cervical spine injuries are due to motor vehicle accidents. Proper extrication of the victims is vital; the ideal device should be easily assembled and applied, should facilitate removal of victims from automobile seats without changing the body's position, must not hinder airway access or the performance of cardiopulmonary resuscitation, must accommodate all types of patients, including children and obese or pregnant patients, and must completely immobilize the patient, especially if hyperextension is suspected. Current methods of immobilization, such as the use of a soft collar and sandbags, allow neck extension; the short board protects against extension but interferes with airway access. Newer devices are discussed in this article. Injuries of the upper cervical spine are less common but more serious than those of the lower portion and usually involve the vertebral arch. Radiologic examination of the first and second cervical vertebrae and the seventh cervical and first thoracic vertebrae should be emphasized. If lateral and anteroposterior views do not reveal abnormal findings and injury is still suspected, oblique views and computed or conventional tomography should be used. Cervical spinal cord injuries can be minimized or prevented if proper early management is applied. PMID- 3046735 TI - Doctors win latest round in BC billing-number dispute. PMID- 3046736 TI - Fine-needle aspiration biopsy of cold thyroid nodules. AB - This study analyzes the results of fine-needle aspiration biopsy (FNAB) of hypofunctioning thyroid nodules performed by one physician. There were 68 patients (age range, 20 to 73 years) with 83 aspirations; 30 were interpreted as positive for neoplasm (adenoma or carcinoma), 43 were negative, and ten (12%) were technically unsatisfactory. Thyroidectomy was performed on 25 patients who had positive aspirates. Subsequent morphologic study showed that 13 patients had carcinomas, ten had adenomas, and two had adenomatoid nodules (false-positive rate of FNAB for neoplasms was 8%). One of three thyroidectomy patients with negative preoperative aspirates had a carcinoma and two had adenomas (estimated minimal false-negative rate of FNAB was 9%). Nineteen patients who underwent thyroidectomies had dynamic radioisotopic thyroid angiography. There was no correlation between the pattern of vascularity and the type of neoplasm. Ultrasound (US) study was performed on 17 patients. Both adenoma and carcinoma can be solid or partially cystic. Although approximately 33% of the nodules initially diagnosed by FNAB as follicular or papillary neoplasms had different interpretations on subsequent examination of thyroidectomy specimens, 93% of the patients selected to be operated on had either adenoma or carcinoma. Thus, in this series, FNAB of cold thyroid nodules gave more useful diagnostic information than nodule size, dynamic radioisotopic scan, or US studies. PMID- 3046738 TI - A murine model to evaluate the ability of in vitro clonogenic assays to predict the response to tumors in vivo. AB - The use of the human tumor cloning assay as a predictor of clinical response of human tumors to drugs is predicated on the hypothesis that the in vivo response of a tumor to a drug can be correlated with the in vitro response of cells derived from the tumor. To test this hypothesis, we utilized a murine tumor model in which the in vivo and in vitro responses of a tumor can be accurately and reproducibly compared. Drug activity was assessed in P388 leukemia with the standard in vivo antitumor assay (i.p. tumor/i.p. drug administration) and an in vitro assay wherein the ascites tumor cells are removed from mice, treated with a drug, and directly cloned in soft agar to measure clonogenic capacity. The response of P388 cells to analogues within four separate classes of antitumor agents, anthracyclines, anthraquinones, platinum(II) coordination complexes, and phosphinogold(I) complexes was evaluated. The clonogenic assay failed to discriminate between highly active in vivo antitumor agents and analogues with only marginal in vivo efficacy (i.e., doxorubicin and daunorubicin versus rhodomycins A and B, ametantrone versus NSC 276740, cisplatin versus transplatin, [Au(dppe)2]Cl versus [Au(depe)2]PF6. Furthermore, the in vitro clonogenic assay failed to detect carboplatin which was a highly active agent in vivo. The basis for these discrepancies was explored by a more detailed comparison of doxorubicin and rhodomycin B. In vivo or in vitro drug exposure with subsequent measurement of cell kill by the in vitro clonogenic and in vivo tumorigenic assay demonstrated that the in vitro assay overestimated the cytotoxic potency of the drugs relative to the tumorigenic assay. Treatment of tumors in vivo with doxorubicin at doses below the maximally tolerated dose in mice resulted in multiple log cell kill as measured in vitro or in vivo, whereas rhodomycin B was cytotoxic only at dose levels exceeding its maximally tolerated dose. The results indicate that a subset of tumor stem cells capable of forming colonies in soft agar are significantly more sensitive to the cytotoxic effects of anthracyclines than are in vivo tumorigenic stem cells. Cytotoxic potency as measured by an in vitro soft agar clonogenic assay is not an accurate predictor of in vivo antitumor efficacy even in a model in which ascites tumor cells are directly exposed to i.p. drug. The in vitro cytotoxicity assay is useful only as a nonselective prescreen and must be used in combination with other indicators of tumor cell selectivity and dose-limiting organ toxicity. PMID- 3046737 TI - Second malignancies after childhood Hodgkin's disease. The Memorial Sloan Kettering Cancer Center experience. AB - A review of the Memorial Sloan-Kettering Cancer Center experience with second malignancies (SM) after childhood Hodgkin's Disease (HD) identified 17 SM in 320 patients who survived more than 1 year from, and were 15 years old or younger at the time of, HD diagnosis (1949 to 1983). Of 254 previously untreated patients, 12 SM were noticed as compared with 0.606 expected on the basis of rates in the general pediatric population (relative risk, 19.8; 95% confidence interval, 10.2 to 34.6). For patients who received multi-agent chemotherapy, the cumulative probability of developing acute nonlymphocytic leukemia (ANLL) or bone sarcoma was 6.2% and 5.5%, respectively, at 10 years from the initiation of therapy; the cumulative risk of all SM in this group reached 18.7% at 15 years. For patients who received radiation alone or with single-agent chemotherapy, the cumulative risk of SM rose from 0% at 10 years and 2% at 15 years, to 10.7% at 25 years from the initiation of treatment. The risk of ANLL after childhood HD was highest in the first 5 to 10 years after combined modality treatment, and aggressive forms of NHL were associated with excessive immunosuppression. Bone sarcomas predominated in solid SM in the first decade after HD treatment, whereas "adult type" cancers, for example, breast and colon carcinomas, were more delayed. Our findings, supported by a literature review, point to a therapy-related enhanced risk of approximately age-appropriate solid SM. This possibility mandates careful surveillance of long-term survivors of childhood HD. PMID- 3046739 TI - Measurement of local Mr 97,000 and 250,000 protein antigen concentration in sections of human melanoma tumor using in vitro quantitative autoradiography. AB - An assay method that uses 125I-labeled monoclonal antibody (MoAb) and in vitro quantitative autoradiography was developed to determine the local concentration of tumor-associated antigens in tissue sections. Human melanoma biopsy specimens were evaluated for the expression of the Mr 97,000 and 250,000 protein antigens using MoAb-96.5 and MoAb-9.2.27, respectively. Tissue sections were incubated in solutions of increasing concentration of 125I-labeled MoAb with or without an excess of unlabeled antibody. Quantitative autoradiography was performed on the sections and compared with 125I standards to determine tumor-bound radioactivity and calculate bound pmol of MoAb per g of tumor. The total binding, nonsaturable binding, and specific binding of 125I-labeled MoAb to tumor were then computed. Specific binding of MoAbs to tumor tissue was saturable in all antigen-positive tumors. The maximal concentration of specific binding of antibody to tissue (Bmax) represented the tissue antigen concentration. Estimates of the Ka of antigen/antibody binding were also made. The reliability of the measurements was confirmed by testing sections from mixtures of antigen-positive and antigen negative cells. PMID- 3046740 TI - Preparation of anti-ras Mr 21,000 protein monoclonal antibodies and immunohistochemical analyses on expression of ras genes in human stomach and thyroid cancers. AB - Sixteen clones (RASK-1 to -16) of murine monoclonal antibodies were raised against ras Mr 21,000 protein (p21). The p21 produced by Escherichia coli with inserted v-Ki-ras genes was used as immunogen. RASK-1 was found to be specific for Ki-ras p21, whereas RASK-2 to -16 reacted with the p21s of Ki-, N-, and Ha ras genes in both enzyme-linked immunosorbent and immunoblotting assays. Binding inhibition assays with biotinylated monoclonal antibodies by enzyme-linked immunosorbent assay showed that the monoclonal antibodies of the 16 clones included those binding to several mutually distinct sites on p21. The expressions of ras p21 in human stomach and thyroid tissues were examined with RASK-3, which reacted with all the Ki-, N-, and Ha-ras p21s immunohistochemically by the avidin biotin peroxidase complex method. Formalin-fixed, paraffin-embedded tissues of 101 cases of stomach cancer, 53 cases of noncancerous stomach, 74 cases of cancer of the thyroid, and 59 cases of noncancerous thyroid were analyzed. In both the stomach and thyroid, cancer cells expressed p21 predominantly. Cells of cases with various noncancerous disorders as well as certain types of normal cells were also p21 positive. These findings suggest that precaution is required in use of p21 as a cancer marker. Expression of p21 was noted in moderately to well differentiated stomach cancer, intestinal metaplasia, and atypical hyperplasia. This finding suggests that the appearance of p21 in stomach cancer may be initiated before cytological transformation. PMID- 3046742 TI - Leucovorin and 5-fluorouracil as a treatment for disseminated cancer of the pancreas and unknown primary tumors. AB - Chemotherapy with leucovorin (100 to 200 mg) and 5-fluorouracil (30 mg/kg) every 2 wk produced four (three complete) objective responses among a group of eight patients with early metastatic pancreatic primary and unknown cancers. Complete remissions were associated with exceptionally long durations of survival, one in a patient failing prior combination chemotherapy. This treatment warrants testing because of its ease, scientific rationale, and the large population of patients with early metastatic pancreatic cancer for whom there is no accepted treatment. Early metastatic disease is defined as small metastatic lesions not immediately life threatening found in a physiologically intact patient. Controlled trials, demonstrating benefit associated with other 5-fluorouracil-containing regimens for patients with nonmetastatic stages of pancreatic cancer, provide a rationale for extending testing of leucovorin and 5-fluorouracil to other early stages of pancreatic cancer. PMID- 3046741 TI - ras oncogenes and phenotypic staging in N-methylnitrosourea- and gamma irradiation-induced thymic lymphomas in C57BL/6J mice. AB - We have investigated stages of thymic lymphoma development in radiation and N methylnitrosourea (NMU)-treated C57BL/6J mice. The lymphoma cell was identified serologically as a cortical population bearing MEL-14hi, H-2Khi, and IL-2R+ surface markers. According to these parameters in C57BL/6J mice the lymphoma cell was the same regardless of inducing agent or activated oncogene (ras or non-ras). Transforming activity in the radiation and NMU-induced tumors was analyzed using both the nude mouse tumorigenicity assay and the focus-forming assay. 8/10 NMU induced tumors and 12/15 radiation-induced tumors showed transforming activity in the tumorigenicity assay. Southern blot analysis of the nude mouse transformants demonstrated K-ras transforming sequences in eight of eight NMU-induced lymphoma DNAs, two of 12 radiation-induced lymphoma DNAs and N-ras transforming sequences in five of 12 radiation-induced lymphoma DNAs. The non-ras transforming activity in five DNAs from radiation-induced thymic lymphomas indicates the presence of an unidentified oncogene(s) in these tumors. Staging of thymic lymphoma development in this animal model system will allow the study of oncogene activation early in the course of carcinogen-induced disease. These results also emphasize the high sensitivity of the nude mouse assay to score for activated oncogenes and might also indicate a high frequency of K-ras activation in NMU-induced lymphomas in C57BL/6J mice. PMID- 3046743 TI - Monitoring of interaction products of cis-diamminedichloroplatinum(II) and cis diammine(1,1-cyclobutanedicarboxylato)platinum(II) with DNA in cells from platinum-treated cancer patients. AB - The formation and stability of interaction products between the anti-cancer drug cis-diamminedichloroplatinum(II) (cis-DDP) and DNA were studied in buccal epithelial and urinary cells from ten cancer patients who received cis-DDP-based therapy. Buccal cells were collected 1 h before and 1-2 h after i.v. infusions with cis-DDP. The interaction products were visualized in an immunocytochemical peroxidase assay, using an antiserum against cis-DDP-modified calf thymus DNA. The nuclear staining density was measured by microdensitometry. Nuclear staining densities in buccal cells after infusions of greater than or equal to 20 mg/m2 cis-DDP were always higher than pretreatment values. Repeated sampling from individual patients treated for 2-5 consecutive days with daily doses of 20-70 mg/m2 cis-DDP indicated that cis-DDP-DNA binding in buccal cells increased in proportion to the cumulative total dose of cis-DDP. The variation in dose-density response between patients was 17%. Apparent adduct loss in buccal cells from four patients, as measured 8-17 days after the last infusion, amounted to 67-86%. Platinum-induced DNA modifications could also be detected in buccal cells from two cis-diammine(1,1-cyclobutanedicarboxylato)platinum(II)-treated patients. In vitro experiments with human buccal cells and lymphocytes indicated linear relationships between DNA modification and either cis-DDP concentration or incubation time. Nuclear staining densities in pretreatment buccal cells from ten cancer patients treated in vitro with 33 microM cis-DDP for 1 h revealed that interpatient variation in in vitro DNA modification by cis-DDP was low. No quantitative correlation was found between in situ and in vitro DNA modification. PMID- 3046744 TI - Clinical trials with human tumor necrosis factor: in vivo and in vitro effects on human mononuclear phagocyte function. AB - The purpose of this investigation was to understand the biological effects of recombinant human tumor necrosis factor used as therapy for cancer. We studied changes in mononuclear phagocyte function following exposure to this cytokine in vitro or in vivo. Tumor necrosis factor increased phorbol myristate acetate induced hydrogen peroxide production 8- to 20-fold in peripheral blood monocytes and peritoneal macrophages in vitro in a dose-dependent manner. Similarly, tumor necrosis factor increased phorbol myristate acetate-induced peroxide production 2.3-fold in monocytes isolated from nine patients following an i.v. infusion of this cytokine (40 to 200 micrograms/m2). In addition, tumor necrosis factor induced a 2.3-fold increase in tissue factor-like activity in mononuclear phagocytes in vitro. In vivo, tumor necrosis factor induced a trend toward higher procoagulant activity in monocytes, although this change was not statistically significant. We also noted a trend toward increased activated partial thromboplastin times and the presence of fibrin D-dimer in patients treated with tumor necrosis factor, demonstrating activation of the coagulation and fibrinolytic systems. Thus, in vivo treatment of humans with i.v. recombinant human tumor necrosis factor induced functional changes in mononuclear phagocytes similar to those noted with in vitro treatment. PMID- 3046745 TI - Structural studies of the Escherichia coli O-149 O-antigen polysaccharide. AB - The structure of the O-antigen polysaccharide from Escherichia coli O-149 has been investigated; methylation analysis, partial hydrolysis with acid, and n.m.r. spectroscopy were the principal methods used. It is concluded that the polysaccharide is composed of trisaccharide repeating-units having the following structure. (Formula: see text). The absolute configuration at the acetalic carbon atom of the pyruvic acid residue is S. PMID- 3046746 TI - Recovery of a cell surface fetal antigen from circulating immune complexes of melanoma patients. AB - A well-characterized 69.5 x 10(3) dalton glycoprotein fetal antigen (FA), isolated from the spent culture medium of a melanoma cell line, UCLA-SO-14 (M14), was utilized to characterize the antigen component of circulating immune complexes (CIC) from melanoma patients. Ten serum samples from five patients with stage II melanoma at 1 and 4 months prior to the clinical detection of recurrent disease were selected for study. The CIC were dissociated with low pH and ultrafiltered through a 100 x 10(3) dalton exclusion limit membrane. The low pH treatment resulted in an increase in antibody titer in eight of ten serum samples. The antibody activity in membrane immunofluorescence was quantitatively inhibited by the filtered antigen fraction and purified FA, suggesting the presence of anti-FA antibodies in the treated serum, which possibly were complexed with FA in the untreated sample. As determined by competitive inhibition in an enzyme-linked immunosorbent assay, the filtrate (antigen fraction) contained an antigen that was immunologically similar to FA. These results clearly demonstrate that FA, expressed on the cell surface of melanoma cells, is present in CIC of selected melanoma patients. PMID- 3046749 TI - Distribution of glutamate decarboxylase-like immunoreactivity in the sixth abdominal ganglion of the cockroach Periplaneta americana. AB - An antiserum against glutamate decarboxylase (GAD) of the rat brain was used to locate GAD activity in sections of the nervous system of the cockroach, Periplaneta americana. The sixth abdominal ganglion was chosen because electrophysiological evidence suggests the presence of GABAergic inhibitory synapses in the cercal-giant interneuron system. Groups of somata and numerous fibres and tracts were positively labelled by the GAD antiserum. A posterior group of labelled somata could be identified close to the entry of the cercal nerves. A line of somata clusters lay along a ventro-lateral furrow. Another discrete row of GAD-like cells was located dorso-laterally. Some small cells among the dorsal unpaired neurons were labelled. A small central group appeared under these cells. An abundance of GAD-like processes and transversal tracts were found within the neuropile. The different systems of GABAergic inhibitors in the ganglion are discussed; in particular we show that the fibres of cercal nerve X are not labelled. This demonstrates that the latter act on the giant fibres via interneurons. We suggest that the group that sends axons into the overlapping region between the cercal nerve and the giant fibre could be the inhibitory interneurons involved in this system. PMID- 3046747 TI - A controlled trial of GL enzyme in the treatment of acute myocardial infarction. AB - GL enzyme (hyaglosidase) is a highly purified component enzyme of hyaluronidase. A therapeutic trial was carried out in the treatment of suspected myocardial infarction among 1,488 patients presenting within 6 h of the onset of symptoms. No significant reduction in mortality at 6 months was observed in the GL group (15.7%) compared with the placebo group (16.4%). Mortality at 2 weeks was also unaffected by treatment (GL 10.3%; placebo 10.9%). PMID- 3046748 TI - Neuronal systems immunoreactive with antiserum to lamprey gonadotropin-releasing hormone in the brain of Petromyzon marinus. AB - The role of gonadotropin-releasing hormone (GnRH) in mammalian reproduction has been studied extensively; however, the role of a structurally different, but related, decapeptide is not well characterized in the most primitive class of vertebrates, Agnatha. Utilizing an antiserum directed to the recently characterized lamprey GnRH, we examined immunoreactive neuronal perikarya and nerve fibers in sections from the brain of the sea lamprey, Petromyzon marinus, using the unlabeled peroxidase-antiperoxidase method. Neuronal perikarya and fibers were immunopositive with antisera generated to lamprey GnRH and also to certain antisera generated to mammalian GnRH. Immunopositive neuronal perikarya were detected in an arc-shaped population extending from ventral to dorsal preoptic areas. Fibers from these cells projected to the neurohypophysis via the preoptico-hypophyseal tract, but in addition also protruded into the third ventricle. Additionally, some fibers coursed along the external surface of the brain, and may also release GnRH into meningeal compartments. The presence of fully processed, mature decapeptide is indicated within neuronal perikarya, as well as in projecting nerve fibers and terminals. No reaction product was detected in sections incubated with an antiserum to the interior amino acid sequences of mammalian LHRH. This finding supports the structure reported for lamprey GnRH by Sherwood et al. (1986). PMID- 3046750 TI - The "trigger factor cycle" includes ribosomes, presecretory proteins, and the plasma membrane. AB - Trigger factor is a soluble, 63,000 dalton protein of E. coli that stabilizes proOmpA, the precursor form of a major outer-membrane protein, in a conformation competent for in vitro membrane assembly. There is approximately one trigger factor molecule bound to each 70S ribosome isolated from cell extracts in physiological buffers. Trigger factor dissociates from ribosomes in 1.5 M LiCl and reassociates with salt-washed ribosomes in low-salt buffer. Binding is exclusively to the 50S (large) subunit, known to contain the exit domain for nascent polypeptide chains. In addition to its associations with proOmpA and ribosomes, excess trigger factor can compete with the proOmpA-trigger factor complex for a limited number of membrane sites that are essential for translocation of proOmpA. These data suggest a model of trigger factor cycling between the cytoplasm, the ribosome, presecretory proteins, and membrane receptor proteins. PMID- 3046751 TI - mab-3, a gene required for sex-specific yolk protein expression and a male specific lineage in C. elegans. AB - The gene mab-3 appears to regulate a subset of sex-specific events in C. elegans male development. Mutations in mab-3 have no apparent effect on hermaphrodites, but cause synthesis of yolk proteins and a limited lineage alteration in males. We infer that mab-3 has at least two distinct male-specific functions. First, mab 3 activity prevents yolk protein production by males, without affecting stage or tissue specificity of expression. Second, mab-3 activity is required for expression of the male V ray cell lineage. Epistasis analysis is most consistent with a model in which mab-3 is controlled by tra-1, the last switch gene known to act in the somatic sex determination pathway. We discuss how genes such as mab-3 might generate sexual dimorphism. PMID- 3046752 TI - Control of the yeast cell cycle is associated with assembly/disassembly of the Cdc28 protein kinase complex. AB - The Saccharomyces cerevisiae gene CDC28 encodes a protein kinase required for progression from G1 to S phase in the cell cycle. We present evidence that the active form of the Cdc28 protein kinase is a complex of approximately 160 kd containing an endogenous substrate, p40, and possibly other polypeptides. This complex phosphorylates p40 and exogenous histone H1 in vitro. Cell cycle arrest during G1 results in inactivation of the protein kinase accompanied by the disassembly of the complex. Furthermore, assembly of the complex is regulated during the cell cycle, reaching a maximum during G1. Partial complexes thought to be intermediates in the assembly process phosphorylate histone H1 but not p40. Addition of soluble factors to these partial complexes in vitro restores p40 phosphorylation and causes the complex to increase to the mature size. A model is presented in which p40 phosphorylation is required during G1 for cells to initiate a new cell cycle. PMID- 3046754 TI - Denturism. PMID- 3046755 TI - Functional properties of collagen and elastin. PMID- 3046756 TI - Crystals in joints. PMID- 3046753 TI - The sequence specificity of homeodomain-DNA interaction. AB - The Drosophila developmental gene, engrailed, encodes a sequence-specific DNA binding activity. Using deletion constructs expressed as fusion proteins in E. coli, we localized this activity to the conserved homeodomain (HD). The binding site consensus, TCAATTAAAT, is found in clusters in the engrailed regulatory region. Weak binding of the En HD to one copy of a synthetic consensus is enhanced by adjacent copies. The distantly related HD encoded by fushi tarazu binds to the same sites as the En HD, but differs in its preference for related sites. Both HDs bind a second type of sequence, a repeat of TAA. The similarity in sequence specificity of En and Ftz HDs suggests that, within families of DNA binding proteins, close relatives will exhibit similar specificities. Competition among related regulatory proteins might govern which protein occupies a given binding site and consequently determine the ultimate effect of cis-acting regulatory sites. PMID- 3046757 TI - Renal biochemistry in rheumatic disease. AB - In patients with osteo-arthritis, renal disease will almost invariably be the consequence of unrelated coincidental renal pathology. In polyarthritic rheumatoid arthritis and ankylosing spondylitis, renal disease can arise as part of the pathology of a systemic disease, or more likely its treatment. As a rule amyloidosis is the most important complication in terms of progressive irreversible loss of renal function. By contrast, SLE, polyarteritis and Sjogren's disease carry a much higher risk of renal involvement. The regular evaluation of renal function in such patients can enable the kidney damage to be assessed at an early stage and allow treatment to be instituted early. PMID- 3046758 TI - Biochemical aspects of infection in rheumatoid arthritis and ankylosing spondylitis. PMID- 3046759 TI - Proteoglycans of joint cartilage. Structure, function, turnover and role as markers of joint disease. AB - Joint cartilage consists of cells embedded in a matrix of fibrous collagen within a concentrated water-proteoglycan gel. The integrity of this matrix is crucial for the biomechanical properties of the joint cartilage. The different components of the matrix are synthesized and degraded by the cartilage cells, a process regulated by the amount of mechanical stress applied to the chondrocytes as well as by peptide factors and hormones present in synovial fluid. The proteoglycans are large macromolecules consisting of a protein core to which are attached multiple chains of glycosaminoglycans and oligosaccharides. During normal and pathological turnover, degradation products are released to the synovial fluid and to the circulation. Newly developed assays allow the sensitive and specific detection of these fragments in joint fluid and serum. Results of experimental and clinical investigations suggest that these assays will be of value in efforts to diagnose, grade and predict the outcome of inflammatory and degenerative joint disease. PMID- 3046760 TI - Biochemistry of bone. PMID- 3046762 TI - Preparation of [3-(2-pyridyldithio)propionoyl]insulins and horseradish peroxidase insulin conjugates by high-performance liquid chromatographic separation. PMID- 3046761 TI - Every ribosomal suppressor mutation in Aspergillus nidulans has a unique and highly pleiotropic phenotype. AB - 18 suppressors of alcR125 have been selected in Aspergillus nidulans. They have been located in genes as follows: 12 in suaA, 1 in suaB and 5 in suaC. Suppressors have been examined to see whether their phenotype is diagnostic for their genotype. Several new traits are described: conidial viability, cycloheximide resistance, fertility, suppression of niaD500, niaD501 and fwA1. These tests, added to those already in use, provide a battery of tests suitable for assigning suppressor mutations to physiological type (tRNA or ribosomal), and in one case to a specific gene since only suaA mutations suppressed fwA1. A very broad range of phenotypes was associated with suppressors such that every mutation had a unique phenotype. This indicates that the ribosomal suppressor mutations are in genes which code directly for ribosomal proteins, rather than genes which code for modifying enzymes. PMID- 3046763 TI - Prolyl endopeptidase from bovine testis: purification, characterization and comparison with the enzymes from other tissues. PMID- 3046765 TI - [Advances in research on the diagnosis and treatment of nonspecific ulcerative colitis with traditional Chinese medicine and Western medicine]. PMID- 3046764 TI - [History of radix Phytolaccae processing]. PMID- 3046766 TI - Effects of cancer treatment on the reproductive system. AB - Many patients with Hodgkin's disease, acute leukemia, non-Hodgkin's lymphoma, testicular cancer, and other tumors now regularly achieve sustained clinical remissions and cures. Drugs used in the treatment of cancer have profound and often lasting effects on the testis and ovary. Germ cell production and endocrine function may both be altered with the magnitude of the effect related to the age, pubertal status, and menstrual status of the patient as well as to the particular drug, dosage, or combination administered. The primary testicular lesion caused by all antitumor agents studied thus far is depletion of the germinal epithelium lining the seminiferous tubules. Combination chemotherapy regimens that include alkylating agents produce germinal aplasia and permanent infertility in the majority of patients. The risk of ovarian injury following combination chemotherapy is clearly related to the age of the patient at the time of treatment. Overall, 40 to 50% of women treated with combination chemotherapy become amenorrheic, although the frequency of amenorrhea in women older than 35 years may be as high as 90%. Interventions to protect the gonads from the effects of chemotherapy have not yet been developed; thus, male patients should be offered an opportunity to store semen prior to treatment and all patients should be counseled concerning the potential gonadal toxicity of cancer chemotherapy. PMID- 3046767 TI - Plasma cell dyscrasias: current status. AB - This is a review of the current status of the monoclonal gammopathies (plasma cell dyscrasias). We begin with the recognition of a monoclonal protein in serum and urine. We briefly discuss the differential diagnosis of the monoclonal gammopathies. Clinical and laboratory findings as well as the management of multiple myeloma are addressed. Future approaches for the treatment of myeloma are provided. The variant forms of multiple myeloma, including smoldering myeloma, plasma cell leukemia, nonsecretory myeloma, IgD myeloma, osteosclerotic myeloma, solitary plasmacytoma of bone, and extramedullary plasmacytoma, are briefly reviewed. Diagnosis and treatment of Waldenstrom's macroglobulinemia are presented. The recognition and differential diagnosis of the heavy-chain diseases (gamma, alpha, and mu) are included. Monoclonal gammopathy of undetermined significance ("benign" monoclonal gammopathy) is presented in detail. Amyloidosis is not included in this review. PMID- 3046768 TI - [Incidence of pathogenic Escherichia coli in acute infantile diarrhea in Dakar]. AB - We have studied the incidence of enteropathogenic (EPEC), enteroinvasive (EIEC) and enterotoxigenic (ETEC) Escherichia coli associated with infant acute diarrhoeal disease in Dakar during a period of one year. We report 405 strains of Escherichia coli suspected to be the etiologic agent of the diarrhoea and isolated from 405 diarrheic stools of 0-5 years old children. We have isolated 119 pathogenic Escherichia coli with 63 EPEC (15.5%), 3 ETEC (0.7%) and 53 ETEC (13.1%) including 23 strains releasing heat-labile enterotoxin (LT+) and 30 strains releasing heat-stable enterotoxin (ST+). No ST+/LT+ strain was isolated. Escherichia coli with colonization factor antigens were isolated from 62 children. Almost all of them are CFAI+. Only one strain is CFAII+ and another one agglutinates with both CFAI and CFAII antisera. Among these CFA+ strains 5 belong to the EPEC group, 29 are enterotoxigenic (25 ST+ and 4 LT+) and 28 do not belong to any known etiopathologic group. Near 70% of the pathogenic Escherichia coli are from infants less than one year old, with a highest frequency between 7 and 12 months. Prevalence of ETEC is higher during the raining season. The existence of a great number of strains that belong to none of the 3 groups of etiopathologic Escherichia coli emphasis the need to search other factors of pathogenicity. PMID- 3046769 TI - [The chloroquine resistance of Plasmodium falciparum in Vanuatu (1980-1984): appearance, evolution, distribution]. AB - Chloroquine resistance of P. falciparum was studied in Vanuatu from March 1981 to July 1984, with limited means, and a non-systematic procedure; it was first evidenced in 1980. In vivo chloroquine resistance criteria were those established by W. H. O. In vitro testing methods were the standard W. H. O. macro-test and micro-test. 124 in vivo chloroquine resistant cases were seen: 111 cases in hospital (63 R III, 28 R II and 20 R I) and 13 cases in field studies (1 R III, 4 R II and 8 R I). 87 in vitro chloroquine sensitivity tests were carried out. A high failure rate was apparently due to a defective batch of lyophilised culture medium. Out of the 25 isolates successfully tested, 22 showed in vitro chloroquine resistance (88%). Correlation between in vivo and in vitro resistance was good in 13 cases studied by both methods. Moreover, 13 in vitro mefloquine sensitivity tests evidenced a high sensitivity to this drug. Chloroquine resistant malaria thus appears to be extended to the whole country. Its prevalence remains unknown but was estimated at 60% at least in 1984. Moreover, a geographical spread of chloroquine resistance from north to south of the group was evidenced between 1980 and 1984, identical to that of the dramatic increase of P. falciparum incidence at the same period of time. Possible mechanisms of the advent of chloroquine resistance in Vanuatu are discussed. PMID- 3046770 TI - An endemic focus of visceral leishmaniasis in Meshkin-Shahr, east Azerbaijan province, north-west part of Iran and IFA serological survey of the disease in this area. AB - The incidence of visceral leishmaniasis has being increased in Iran during the recent decade. Since 1980, more than 200 cases have been diagnosed from East Azerbaijan province, mostly, from Meshkin-Shahr area. It seems, that kala-azar has being endemic in this area for a long time. The majority (86%) of kala-azar cases were found among children up to 4 years. The sex incidence ratio of males/females was 1.27/1. In IFA serological survey, sero-positive rate in females was higher than males. However, geometric mean of leishmanial antibody titers in males was, slightly, more than females. These serological findings indicate that females are exposed to the infection at least as much as males. The cross-sectional IFA serological survey, relatively reflected the kala-azar status among different studied groups with various incidences of the disease in Meshkin Shahr area. IFAT showed also a good efficiency in the assessment of the treatment in the treated kala-azar patients. PMID- 3046771 TI - [Toxoplasmosis in Niger. A serological analysis of 400 subjects]. AB - A toxoplasmosis serological study using an indirect immunofluorescence method was made in 400 subjects in the Republic of Niger because the epidemiologic findings of this disease were very poor. Serum antitoxoplasmic antibodies were present in 18.2%. In sahelian areas prevalence of such toxoplasmic antibodies is low in the majority of studies. Analysis of seropositivity and age pointed out a late seroconversion. PMID- 3046772 TI - [Ivermectin and human onchocerciasis. A study of 234 onchocerciasis patients in the Republic of Mali]. AB - In a double blind study Ivermectin has been compared to a placebo in 234 male and female with onchocerciasis who had more than 20 microfilariae per milligram of skin and moderate ocular involvement. Patient was randomized to receive a simple oral dose of Ivermectin 100, 150 and 200 micrograms/kg or placebo. The following was 12 months. The decrease of microfilarodermia since to the 3rd day was from 72.8 to 79.3% of initial rate. At six months it was more than 91% and more than 87% in 12 months. Ocular microfilariae, initially between 12 and 23 stay lower than 2 at 12 months. Punctuated keratitis disappear and did not recidive still 6 months in patients with persistent microfilariae. Ivermectin produce only few side effects. Negative waves have been observed on ECG but without any clinical signs. The Power efficient dose seen to be 100 micrograms/kg. PMID- 3046774 TI - Intravenous digital subtraction angiography for preoperative evaluation of patients with extensive renal calculus disease. PMID- 3046773 TI - Some thoughts on osteoporosis in women. PMID- 3046775 TI - Bronchial hyperresponsiveness is not bronchial hyperresponsiveness is not bronchial asthma. PMID- 3046776 TI - Comparison of cellular and protein changes in bronchial lavage fluid of symptomatic and asymptomatic patients with red cedar asthma on follow-up examination. AB - Seventeen patients with occupational asthma due to western red cedar had bronchial lavage during follow-up examination after removal from exposure for at least 1 year. Seven patients were asymptomatic while ten continued to have symptoms of asthma requiring treatment. Symptomatic patients had evidence of airway inflammation, as reflected by a significantly higher total cell count, neutrophils and eosinophils, as well as an increase in protein and albumin in their bronchial lavage fluid compared to those without symptoms. Asymptomatic patients had no evidence of airway inflammation in the lavage fluid. There was no correlation between the degree of non-specific bronchial hyperresponsiveness and the number or percentage of inflammatory cells to suggest that cellular infiltration is the sole cause of persistent bronchial hyperresponsiveness. PMID- 3046777 TI - Identification of allergens and antigens of Bermuda grass (Cynodon dactylon) pollen by immunoblot analysis. AB - Allergens and antigens of Bermuda grass pollen fractionated by SDS-polyacrylamide gel electrophoresis and transferred to nitrocellulose membranes were identified using twenty-one sera of Bermuda grass pollen-allergic patients. The IgE- and IgG binding pollen components transferred to nitrocellulose were detected by reaction with enzyme-labelled anti-human IgE and anti-human IgG, respectively. There was heterogeneity in both IgE- and IgG-binding patterns of the allergic sera tested. Fourteen pollen components, ranging in molecular weight from 16,000 to 88,000 daltons, bound to IgE antibodies. Only two of the fourteen allergens identified reacted with IgE antibodies of more than 50% of the twenty-one allergic sera. The pollen component with a molecular weight of 32,000 daltons showed by far the highest frequency of IgE binding, being recognized by sixteen (76%) of the twenty one sera examined. Fifteen (71%) of the twenty-one sera tested had IgE antibodies that reacted with more than one of the fourteen allergenic components identified. Pollen components recognized by IgE antibodies also reacted with IgG antibodies, and there were components only recognized by IgG antibodies. Results obtained from this study should be useful both clinically and in research. PMID- 3046778 TI - Protein C in human plasma determined by homogeneous enzyme immunoassay with use of a centrifugal analyzer. AB - We describe the simple and rapid enzyme immunoassay of protein C in human plasma with use of a Cobas Fara centrifugal analyzer. The antibody, labeled with horseradish peroxidase, is reacted with antigen (protein C) for 15 min. The peroxidase activity of the resulting antigen-antibody conjugate is measured at 500 nm for 5 min in the presence of excess H2O2, phenol, and 4-aminoantipyrine, as compared with that of free conjugates. Results are calculated from a stored standard curve and expressed as a percentage of the value determined for a pooled specimen of normal adult plasma. The standard curve is linear from 0% to 200%. The CV is generally less than 4% for different concentrations of protein C. In liver cirrhosis, hepatocellular carcinoma, therapy with warfarin, thrombosis, and disseminated intravascular coagulation, protein C concentrations are about 40-70% of normal. Results obtained with the present homogeneous enzyme immunoassay correlated well with those by enzyme-labeled immunosorbent assay (r = 0.97). PMID- 3046779 TI - Solid-phase enzymoimmunoassay of estrone in serum. AB - This rapid solid-phase enzymoimmunoassay for estrone in serum or plasma is done on microtiter plates after the serum is extracted with diethyl ether. No chromatographic or centrifugation steps are involved. The detection limit of the assay is 380 fg per well (28 pmol/L). Intra- and interassay coefficients of variation for the assay of low, medium, and high concentrations of estrone in plasma were respectively 4.4, 9.3; 2.3, 9.1; and 2.0, 6.3 percent. There was a good correlation (correlation coefficient 0.95) between estrone concentrations measured with this assay and with a commercial radioimmunoassay. The analytical procedure is simple, and one person can assay 80 serum samples per working day. We conclude that the assay is very suitable for serum estrone measurements and is more convenient than published radioimmunoassays. PMID- 3046780 TI - Improved agarose electrophoretic method for separating alkaline phosphatase isoenzymes in serum. AB - A modified agarose electrophoretic system for the separation of alkaline phosphatase (ALP, EC 3.1.3.1) isoenzymes is described. Bone, liver, high molecular-mass, and intestinal ALP are separated with high reproducibility. The sensitivity of the agarose system is superior to cellulose acetate in detecting high-Mr ALP. Correlation is good between bone ALP fractions scanned before and after treatment with neuraminidase. Immunoglobulin-bound ALPs, the ALP lipoprotein-X complex, and the additional ALP fraction observed in transient hyperphosphatasemia in children are detected by their peculiar electrophoretic mobility in the proposed system. Approximately 25% of the samples contained an additional fraction of intestinal-type ALP, as evidenced by neuraminidase treatment and use of polyclonal and monoclonal antibodies. Because the electrophoretic mobilities of this "intestinal variant" and of some immunoglobulin-bound ALP fractions are identical to those of bone and intestinal ALP, respectively, treatment of the samples with a polyclonal antibody that reacts with intestinal ALP is advised. PMID- 3046782 TI - Skin-puncture and blood-collecting technique for infants: update and problems. AB - This is updated information on acceptable practice in skin puncture and blood collection in infants, as well as on the devices used, with the additional aim of emphasizing major problem areas and some tentative solutions. Consensus standards for skin puncture have little experimental support, and evade the hard fact that studies are needed to clarify optimum sites for puncture and depth and width of lancets, and to assess the effects of compression and skin resistance in the puncturing process. Preliminary data revealed that the puncturing depth of 2.4 mm recommended for the newborn is excessive. In four of 14 newborns at necropsy, the distance from posterior planar skin surface to underlying bone ranged between 2.0 and 2.2 mm. An experimental lancet, with a 1.8-mm tip length and a diameter of 0.79 mm yielded customary blood volumes from newborns in three of the four pediatric centers where it was tested. Lack of success with the lancet was attributed to inexperienced phlebotomists, not to the lancet's decreased size. Also reviewed are problems with common devices used, and the need for examining the "economy" of blood collection. PMID- 3046784 TI - Texas Section AACC. PMID- 3046783 TI - Macro-creatine kinase co-migrating with CK-MB on electrophoresis. PMID- 3046781 TI - A commercial enzyme immunoassay method (EMIT) compared with liquid chromatography and bioassay methods for measurement of chloramphenicol. AB - A new enzyme immunoassay method (EMIT; Syva Co.) was compared with conventional high-performance liquid chromatography (HPLC) and agar-diffusion bioassay methods for measurement of chloramphenicol in human serum. Forty-nine serum samples were assayed by each of the three methods. Excellent correlation was observed between values by EMIT and by the two conventional methods (r = 0.986 and 0.961). Precision was acceptable (CV less than 5%) with EMIT. Assay of samples containing chloramphenicol glucuronide and chloramphenicol succinate demonstrated that EMIT recognizes only the biologically active (base) form of the drug. The capability to test serum samples as small as 0.2 mL, adaptation to widely available instrumentation, and provision of rapid results are principal advantages of the EMIT method for routine chloramphenicol measurements. PMID- 3046785 TI - Enzymic methanol determination: toxic concentrations of ethanol may give positive values. PMID- 3046786 TI - Erroneous test results obtained with the Tandem-E TSH kit. PMID- 3046787 TI - An enzyme antigen immunoassay for the determination of neuron-specific enolase in serum samples. AB - A method is presented for the detection and quantification of neuron-specific enolase (NSE) in serum samples. It is an enzyme antigen immunoassay (EAIA), relying on specific antibodies to 'catch' the enzyme on a solid support (ELISA plate) whereafter the enzymatic activity of the immunocaptured enzyme is determined, by coupling the reaction to lactate dehydrogenase. The oxidation of NADH to NAD is followed at 340 nm in an ELISA photometer. The method is proven to work well and is able to measure the low amounts of NSE present in serum samples from normal individuals. It does not require labelling of neither antigen nor antibody, and is therefore superior to the commercially available radioimmunoassay (RIA). The method will also work with monoclonal antibodies towards the enzyme as demonstrated by preliminary observations. PMID- 3046789 TI - Coronary artery bypass surgery in women. AB - Coronary bypass surgery is performed more frequently in men than in women. A selection bias in favor of men may exist in currently utilized evaluation precesses for patients with both chest pain syndromes and documented coronary artery disease. Surgery should be considered in women with significant left main coronary artery stenosis, "left main equivalent" coronary disease, severe three vessel coronary disease with/without left ventricular dysfunction, two-vessel coronary disease (including a proximal left anterior descending artery stenosis), and unstable angina pectoris with decreased left ventricular function. Women and men undergoing coronary bypass surgery seem to benefit from internal mammary artery grafts used alone or in combination with saphenous vein grafts. Surgical mortality, incomplete revascularization, early and late graft occlusion, and recurrent angina are more prevalent in women who undergo surgery. However, long term mortality following surgery is similar in men and women. PMID- 3046788 TI - Diagnosis of Zellweger syndrome by rectal biopsy: immunoblot of peroxisomal beta oxidation enzyme and activity of dihydroxyacetone phosphate acyltransferase in rectal mucosa. PMID- 3046790 TI - Association of coronary calcification with obstructive disease in coronary arteries in Indian patients. AB - A total of 1150 consecutive patients (1052 males and 98 females; age 51.2 +/- 10.1 years) with suspected coronary artery disease (Group I) were subjected to fluoroscopy for detection of coronary artery calcification (CAC) and coronary angiography. Another group (Group II) of 120 patients (95 males and 25 females; age 51.4 +/- 9.4 years) catheterized for cardiac diseases other than coronary artery disease (CAD) were subjected to the same protocol of fluoroscopy and coronary angiography to exclude incidental CAD in view of their age. CAC was present in 240 patients (20.0%) in Group I. Of these, 200 (83.4%) had triple vessel disease (TVD); 20 (8.3%) had double-vessel disease (DVD); 19 (7.9%) had single-vessel disease (SVD); and 37 (15.4%) patients had left main coronary disease (LMCAD). Only one of these patients had insignificant CAD considered as "normal" coronary arteries (NC). Incidence of LMCAD, TVD, DVD, SVD, and NC in patients without CAC was 4.4%, 56.3%, 18.2%, 14.0%, and 11.5%, respectively. Incidence of CAC in patients with LMCAD, TVD, DVD, SVD, and NC was 48.1%, 28.1%, 10.8%, 13.0%, and 1.0% respectively. In Group II (n = 120), 24 patients (20%) had CAD, CAC was present in 5 patients with CAD (20.9%), and in two patients without CAD (2%). CAC is relatively uncommon in Indian CAD patients. Its presence, however, indicates severe multivessel disease. PMID- 3046791 TI - Monotherapy of hypertension with darodipine: a new calcium-channel blocker. AB - Calcium-channel blockers are increasingly used as single agents for the treatment of essential hypertension. Following three weeks of single-blind placebo therapy, 43 patients with essential hypertension were randomized into four groups. Group 1 (10 patients) received placebo twice a day, Group 2 (13 patients) received darodipine (PY 108-068) 50 mg twice a day, Group 3 (9 patients) received darodipine 100 mg twice a day, and Group 4 (11 patients) received darodipine 150 mg twice a day. Patients were seen in the clinic weekly for 4 weeks. Clinical and laboratory evaluations were done on each patient. Darodipine caused a sustained decrease in the supine and standing systolic and diastolic blood pressure (p less than .001) and there were no significant pressure differences between the three drug dosages. The effects of the drug on heart rate were not consistent. Placebo had no effect on either blood pressure or heart rate. Side effects were few, mild, and consisted of headaches and peripheral edema, and did not necessitate discontinuation of the drug. No metabolic abnormalities were seen with either low or high doses of the drug. We conclude that: (1) darodipine is effective, safe, and well tolerated; (2) its antihypertensive effectiveness is similar at high and low doses, although the higher doses seemed to have a slightly greater effect on the diastolic arterial pressure; (3) the low dose may be preferable since side effects were dose related. PMID- 3046792 TI - Arthur M. Master, 1895-1973. PMID- 3046793 TI - Antenatal and perinatal causes of handicap: definitions and size of the problem. AB - The conditions of childhood that follow prenatal or perinatal problems are defined. These conditions are mental retardation in varying degree, the cerebral palsies, the syndrome of minimal cerebral dysfunction, language disorders and defects of hearing and vision. The difficulties in accurately measuring disability and handicap are discussed and an estimate of the size of the problem is offered. The contribution made by the low birthweight group is also considered and it is pointed out that, although within this group there is a much higher incidence of problems, the majority of children with handicapping conditions were of normal birthweight. PMID- 3046794 TI - Intrauterine growth retardation and disability. PMID- 3046795 TI - Abnormal antepartum fetal heart rate patterns and subsequent handicap. AB - In this paper data on abnormal antepartum FHR patterns are related to the state of fetal oxygenation, fetal brain abnormalities and to neurological outcome. It is concluded that in IUGR fetuses changes in heart rate (and movement) patterns are late signs of impairment. Antepartum heart rate decelerations are usually the first of the abnormalities detected and are associated with fetal hypoxaemia. A fixed or flat FHR pattern might be indicative of congenital malformations of the brain or of prenatally acquired encephalopathy. Several studies have shown that antepartum FHR abnormalities (usually late decelerations) are associated with an increased risk of subsequent handicap. This risk is related to the degree of FHR abnormality, and especially applies to infants born preterm and/or growth retarded. Late (ante partum) FHR decelerations seem to be more important than 'asphyxia' at birth in determining (neonatal) neurological outcome. In IUGR hypoxaemia is probably associated with deprivation of other nutrients, and thus brain damage in these infants is more likely to be due to chronic malnutrition (including hypoxaemia) than to hypoxaemia alone. This reasoning is supported by morphological findings in IUGR infants. In general fetuses should be delivered before antepartum signs of hypoxaemia appear. Doppler blood--velocity waveform analyses of fetal vessels may detect fetuses at risk for antepartum decelerations, but until now there has been insufficient information about false positive abnormal velocity waveforms to depend absolutely on these. Furthermore, delivery at an earlier age may increase the risk of other neonatal complications. PMID- 3046796 TI - Abnormal antenatal ultrasound findings and subsequent handicap. AB - The modern ultrasound technique in its various modes, real-time B-mode, M-mode, continuous and pulsed wave Doppler ultrasound, makes it possible to study in detail the fetal anatomy and function in utero. Fetometry allows for the evaluation of fetal size and growth and for detecting growth-retarded or macrosomic fetuses. Ultrasound seems, at present, to be the best method available in the detection of the growth-retarded fetus who is at risk of developing perinatal complications and subsequent handicap. Ultrasound can also be used when fetal growth retardation is suspected on clinical grounds. An absolute prerequisite for the proper use of ultrasound fetometry is the estimation of gestational age in early pregnancy. The high resolution of modern ultrasound scanners allows the antenatal detection of even minor fetal structural abnormalities. When lethal malformations are detected early in pregnancy, selective termination of pregnancy can be considered. Fetal abnormality detected in late pregnancy allows for optimal timing and mode of delivery leading to improved management and outcome, thus lowering the risks of subsequent handicap. Ultrasound examination of the extrafetal structures, e.g. umbilical cord, placenta and amniotic fluid volume, may add valuable clinical information. The finding of severe oligohydramnios is associated with increased perinatal mortality and morbidity. Fetal circulation can be evaluated by using the combination of real-time and pulsed Doppler ultrasound or, alternatively, by employing continuous wave Doppler ultrasound alone. Pathological changes in the blood velocity waveforms recorded from the fetal and umbilical arteries may signify very early signs of fetal hypoxia. The method has the potential of becoming a useful clinical method for fetal surveillance. The guidelines for a proper application in the perinatal medicine of the Doppler method have, however, not yet been established. In fetuses with cardiac arrhythmia and/or cardiac malformation, the Doppler investigation of the fetal circulation can be used to evaluate the haemodynamic alterations and, in cases of intrauterine treatment, to supervise and monitor the therapeutic effects. Fetal motor function can be followed and quantified with real-time ultrasound. Abnormal motor activity might indicate bad perinatal outcome. The predictive capacity of the test is increased when several variables are combined (e.g. the fetal biophysical profile score).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3046798 TI - Intrapartum asphyxia and subsequent disability. PMID- 3046797 TI - Cerebral handicap in full-term neonates related to the mechanical forces of labour. AB - Sometimes the relationship between peripartum events and neonatal CNS injury is obvious: for example, following complete abruptio placentae or umbilical cord prolapse and occlusion with a delay of many minutes before delivery of the baby. These circumstances are, of course, rare in modern obstetrics. Usually, when a neonate develops neurological injury, a host of various potentially adverse peripartum factors are assumed to be the aetiology, but without definitive evidence. Among these latter factors are those we have focused on in this paper: the mechanical forces exerted on the fetal head during labour when the full-term fetus is in cephalic presentation. The mechanical events during the first stage of labour are reviewed, showing how uterine contractions result in cervical dilatation and descent and rotation of the fetal head. The consequences of these forces on the fetal intracranial pressure and blood flow are discussed: FHR remains normal up to a certain pressure threshold, above which decelerations occur. In other words, excessive pressures applied to the fetal head, either spontaneously (e.g. uterine tetany) or iatrogenically (e.g. traumatic forceps delivery or excessive fundal pressure) can increase fetal intracranial pressure to such a degree as to result in significant decreases in cerebral blood flow that are associated with fetal heart rate decelerations. Even when decelerations are simultaneous to contractions, decelerations cannot be considered as reflex and innocuous, as they are indeed associated with a decreasing cerebral blood flow. They must therefore be considered and evaluated in the management of labour. Cord compression and functional modifications of intervillous space by mechanical forces may further compromise the biological status of the fetus, leading to severe asphyxia. Neurological evaluation of the neonate within the first few days after delivery is currently the only way to provide the obstetricians with information on the possible consequences of an abnormal labour. The assessment of normality of the CNS in the neonate born at term, and its value in predicting late outcome are discussed. When abnormalities are detected after one or repeated assessments, abnormal neurological signs and symptoms are classified into three grades at the end of the first week. According to our data, a good correlation exists between this neonatal grading of cerebral dysfunction and late outcome. A careful evaluation of fetal head deformation, extensive caput succedaneum, and extensive retinal haemorrhages can help to interpret an abnormal labour retrospectively.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3046799 TI - Breech delivery and mental handicap. AB - Recent studies with strict management protocols for selection of cases for trial of vaginal delivery for full-term breech presentations show no significant differences in outcome brought about by the birth route. This is in clear contrast to some of the older studies which indicated considerable risk for the vaginally born breech infant. Prospective follow-up studies and carefully matched controlled studies with sophisticated neurological evaluations indicate that breech infants, regardless of mode of delivery, will score slightly less favourably than infants born in vertex presentation. This difference seems to reflect prenatal factors rather than birth injuries. PMID- 3046800 TI - Birth trauma and brain damage. AB - Birth trauma is a rare primary cause of perinatal death, occurring at most only once in every 1000-2000 births. As a cause of brain damage and later handicap it is often difficult to dissociate injury at birth from the concomitant effects of asphyxia, growth retardation or preterm delivery. A continuum of reproductive casualty has been postulated, but for trauma is not proven. Among children with cerebral palsy and severe mental retardation trauma may be implicated in a few cases, possibly 1-2 of 1000 deliveries. Vaginal breech delivery has been related to a higher incidence of minimal brain damage syndromes and some of this damage probably has its origin in perinatal trauma. The pregnancies where there is particular risk of birth trauma include those where the infant is large for gestational age, has intrauterine growth retardation, is delivered preterm, is vaginally delivered in breech presentation or is from multiple gestation. Particular care must be given to diagnosis and preventive measures in these cases and competent handling is required if the disaster of brain damage caused by traumatic birth is to be minimized. PMID- 3046801 TI - Developmental neurology of the fetus. AB - The use of modern ultrasound equipment has opened the way to fetal neurology. The aim is to provide the obstetrician with standardized and comprehensive methods to evaluate the neurological condition in compromised fetuses. Such possibilities will greatly improve assessments which have so far been based only on fetal heart rate monitoring and quantitative counts of fetal motility. The most important candidates for neurological assessment are the quality of fetal movements, in particular of general movements, and in the near-term fetus the organization of behavioural states. Less suitable, because of limited sensitivity, is the quantitative assessment of fetal movements, even when carried out with ultrasound. The most problematic approach is testing the fetus with vibro acoustic stimuli, which induce abnormal behavioural states of prolonged duration and are therefore potentially harmful. PMID- 3046802 TI - Perinatal hypoxic-ischaemic brain damage and intraventricular haemorrhage. AB - It is now apparent that the principal lesions of perinatal asphyxia--cerebral hypoxic-ischaemic damage and IVH--are pathogenetically interrelated, a fact that has long been suspected by pathologists (Pape and Wigglesworth, 1979). Prevention of such lesions would require, on the one hand, circulatory support to prevent hypotension and, on the other hand, the means to avoid excessive arterial pressure peaks in the early neonatal period together with experimental and clinical research to determine the optimal PCO2 level. It may therefore be concluded that neonatal ischaemia is a critical determinant for later neurological and intellectual development, and is probably the most important single factor known at present. Our consistent finding of hypoperfusion and, by inference, low metabolic activity supports the hypothesis that this ischaemia may cause minor structural changes in the white matter border zones between major arterial territories, leading to severe learning disorders in these children, who were examined with the 133Xe-inhalation computerized emission tomography technique (Lou et al, 1984). PMID- 3046803 TI - Intrauterine growth retardation and mental handicap: epidemiological evidence. PMID- 3046804 TI - Avascular necrosis and the blood supply of the femoral head. AB - Postmortem femoral artery perfusion revealed abnormalities in the femoral head pattern of vessels in steroid-treated renal transplant patients as compared with controls. Degenerative changes were found in some of the arteries and arterioles of the hip capsule and femoral head in the renal transplant patients. Two of these patients' femoral heads showed microfocal avascular necrosis. Although high dose steroids were used in these patients, none had suffered clinically from hip disease while alive. These findings suggest the possibility that local arterial disease may be involved in the pathogenesis of avascular necrosis. PMID- 3046805 TI - Complete replacement arthroplasty of the hip by the ring ring prosthesis. By P.A. Ring, 1968. PMID- 3046806 TI - Revision of failed total hip arthroplasties with uncemented porous-coated anatomic components. AB - This series represents a relatively short follow-up study of patients who were treated with cementless revisions for failed previous arthroplasties. Many of these cases required extensive bone grafting to the acetabulum and often to the femur. Despite extensive bone grafting, there were no infections. There has been but a single graft resorption after a hemiarthroplasty conversion for recurrent dislocations. To date, all other grafts have remained intact and have shown signs of union. Even though the acetabular components were not anchored in place by adjuvant fixation devices such as screws, migration of the acetabular component has not been a problem. All other components have remained stable, and the supporting grafts appear to have united successfully. Femoral revision has been more technically demanding because the largest stem possible should be placed within the femur to prevent subsidence and provide good stabilization in the proximal metaphyseal area. These short-term results compared favorably with similar series of cemented revisions. Patient selection is important and there are definite candidates for cemented femoral components, particularly with first time revisions in elderly patients. If there is massive osteolysis in the femur, cemented revision is probably not indicated. Long stems should not be used unless necessary. Cortical defects at the tip of the standard stem obviously would require bypassing the stress riser with a longer stem. If, however, the cortex is intact in this region and stability can be achieved, revision should be carried out with a relatively short stem. Techniques for cementless revision are demanding, but with meticulous attention to detail and technical perfection, the method has a most encouraging prognosis. Longer follow-up evaluations will be necessary to make an accurate evaluation of graft incorporation, but short-term results are encouraging to both surgeons and patients. PMID- 3046808 TI - Streptococcal pharyngitis. Comparison of latex agglutination and throat culture. AB - Despite its imperfections, the throat culture remains the "gold standard" against which all rapid streptococcal antigen detection tests are compared. Using triple throat swabs, the accuracy of a rapid latex agglutination (LA) test and back up throat culture was determined and compared with a simultaneously obtained additional throat culture in children with suspected streptococcal pharyngitis. Although there was a 95 percent concordancy between throat cultures, the sensitivity of the throat culture was only 87 percent. Despite the LA test's lower sensitivity (78 percent), in this clinical population with a relatively low prevalence of positive throat cultures (19 percent), the predictive value of a negative LA test was only slightly lower than that of the throat culture (94-95 percent vs. 97 percent). Backup throat cultures are commonly recommended for patients with initially negative LA test results, but 10 percent of the patients with group A beta-hemolytic streptococci-positive throat cultures would have been undetected using this approach. PMID- 3046807 TI - Hemangiomatosis of the colon and peritoneum: case report and management discussion. AB - A 3-month-old infant presented with ascites, anemia, minimal rectal bleeding, and thrombocytopenia. He was found to have several hundred small cavernous hemangiomas of the colon and peritoneal surfaces. Treatment consisted of laser surgery, subtotal colectomy, and steroids. A brief review of the literature on intestinal hemangiomatosis is included as well as the complications of the disease and current therapy. PMID- 3046810 TI - Pediatric medication-induced focal esophagitis. Case report and review. PMID- 3046809 TI - Trisomy 22 mosaicism with normal blood chromosomes. Case report with literature review. AB - A female infant with growth failure, microcephaly, hypertelorism, epicanthal folds, preauricular pit, congenital heart defect, hypotonia, and delayed development is reported. Trisomy 22 mosaicism (46,XX/47,XX,+22) was found in cultured skin fibroblasts but not in blood lymphocytes. Trisomy restricted to skin fibroblasts is uncommon. PMID- 3046811 TI - Cystic fibrosis: enhanced theophylline metabolism may be linked to the disease. AB - Theophylline disposition (5.5 mg/kg administered intravenously) was studied in 12 patients with cystic fibrosis (CF) and 16 healthy control volunteers. Dietary controls and logs were used to minimize the influence of food on theophylline metabolism. Control subjects were restudied in random order on two subsequent occasions after 2 weeks of either pancreatic enzymes or placebo. Theophylline and its three main metabolites, 1-methyluric acid, 3-methylxanthine, and 1,3 dimethyluric acid, were analyzed in serum and urine by HPLC. The total body clearance, renal clearance, nonrenal clearance, and volume of distribution of theophylline were significantly greater (p less than 0.05) in patients with CF than in control subjects. The increased nonrenal clearance was the result of increased biotransformation to each of the three main metabolites. Patients with CF exhibited enhanced N-demethylation and 8-hydroxylation of theophylline, pathways that appear to be mediated by two different families of P-450 enzymes. Theophylline clearance after 2 weeks of pancreatic enzyme administration in the control subjects was the same as with placebo. Possible reasons for enhanced theophylline biotransformation in CF are discussed. PMID- 3046812 TI - The effect of clonidine on the cessation of cigarette smoking. AB - The effect of clonidine on smoking cessation was studied by randomly assigning 186 smokers in a double-blind fashion to either placebo or clonidine. Abstinence from smoking was reported more frequently by subjects receiving clonidine, but the difference was statistically significant only at the end of the first week (34.4% vs 21.5%; p less than 0.05). Bothersome side effects were common and resulted in the early discontinuation of the study medication by 23 of the subjects taking clonidine and eight taking placebo (p less than 0.05). Although this study did not demonstrate a significant effect of clonidine on smoking cessation, a beneficial trend was detected and therefore further trials with transcutaneous delivery of this agent in combination with behavior modification techniques are warranted. PMID- 3046813 TI - Influence of posture on hepatic perfusion and the presystemic biotransformation of propranolol: simulation of the food effect. AB - Several research groups have reported that the oral administration of propranolol with protein-rich food leads to a marked increase (mean + 60%) in the area under the drug plasma concentration-time curve (AUC oral) of this highly metabolized and well-absorbed drug. It has been postulated that this "food effect" is caused at least in part by a transient increase in hepatic blood flow (QH) with its associated decrease in first-pass metabolism (hepatic extraction is a monotonic decreasing function of QH). A randomized crossover study using postural manipulations to produce changes in QH of the magnitude observed after food consumption (20% to 50%) was performed in an attempt to isolate the contribution of transient changes in QH to the food effect phenomenon. A solution of 80 mg propranolol HCl was taken orally and subjects were randomly assigned to postural manipulation protocols that should change QH such that AUC oral would be minimized (phase 1) or maximized (phase 2). Estimated QH (indocyanine green total body clearance from blood) was determined before and at three time points during each phase. It was observed that indocyanine green total body clearance during periods of standing was 15% to 40% below that observed during periods of seating (significant at p less than 0.05 for many of the appropriate comparisons). However, AUC oral for propranolol was not affected (mean +/- 1 SD; AUC phase 2/AUC phase 1+= 0.98 +/- 0.28) by these changes in QH, which are comparable to those encountered after food consumption.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3046814 TI - Comparison of the effects of guanabenz and hydrochlorothiazide on plasma lipids. AB - The effects of hydrochlorothiazide (HCTZ) and guanabenz monotherapy on blood pressure and serum lipoprotein levels were compared in a 14-week, randomized, parallel, double-blind multicenter study of 218 outpatients with mild hypertension. Mean supine blood pressure decreased 13/9 mm Hg in the guanabenz group and 17/11 mm Hg in the HCTZ group, changes that were significantly (p less than 0.01) different from baseline but not significantly different between the two treatment groups. Significant (p less than 0.01) mean decreases in total cholesterol and low-density lipoprotein (LDL) cholesterol levels (of 9 mg/dl and 4 mg/dl from baseline values) occurred during guanabenz treatment; HDL cholesterol levels fell by an average of 4 mg/dl. In the HCTZ group, triglyceride levels were significantly (p less than 0.01) increased by 13 mg/dl, and HDL cholesterol levels fell by 2 mg/dl. The change in LDL cholesterol levels, but not HDL cholesterol levels, was significantly different between guanabenz and HCTZ periods. The results show that guanabez, although providing effective blood pressure control that is comparable to that of HCTZ, has more favorable effects on lipoproteins. PMID- 3046815 TI - Systemic absorption of topically applied ocular atropine. AB - We quantitated atropine plasma levels and monitored blood pressure, heart rate, and salivary secretion after ocular application. Eight patients received 40 microliters 1% atropine in the lower cul-de-sac of one eye in connection with ocular surgery. Atropine plasma levels were determined for 90 minutes by radioreceptor assay. The peak plasma atropine concentration of 860 +/- 402 pg/ml was reached within 8 minutes in all patients. The ocular absorption of atropine was at least as rapid as that reported for intramuscular administration. Ocular atropine did not affect patients' blood pressure or heart rate when compared with those of the placebo group. Thirty minutes after administration of atropine eyedrops, the salivary secretion in the experimental group was reduced, but was statistically insignificant from the placebo group. PMID- 3046817 TI - Pancreatic effect of a hypoglycaemic fragment of human growth hormone (hGH 6-13). AB - The pancreatic effect of a hypoglycaemic fragment of human growth hormone containing the amino acid sequence Leu-Ser-Arg-Leu-Phe-Asp-Asn-Ala (hGH 6-13), was investigated. In partially pancreatectomized rats, hGH 6-13 (3 mg/kg body weight) enhanced glucose utilization in blood as demonstrated in intravenous glucose tolerance tests (IVGTTs). However, the basal levels of plasma insulin in the animals were apparently not affected by acute administration of the hGH fragment and only slightly modulated with prolonged hGH fragment treatment. Direct studies with the isolated pancreatic islets from normal and hGH 6-13 treated rats showed that hGH 6-13 did not influence in vitro or ex vivo insulin release in the absence of glucose but significantly potentiated the glucose induced insulin secretion of the pancreatic islets from treated animals. An increase of 42% in glucose oxidation of the isolated pancreatic islets after exposure to hGH 6-13 was observed. This study reveals significant differences in the molecular mechanism of the hypoglycaemic action between the hGH fragments and orally active sulphonylureas. The findings suggest that the hypoglycaemic hGH fragments, structurally unrelated to sulphonylureas, could be a new group of effective agents to achieve blood glucose normalization without the risk of hyperinsulinaemia, as their pancreatic effect is glucose-dependent. PMID- 3046816 TI - Cardiovascular effects of caffeine and nifedipine. AB - Because caffeine and nifedipine may have opposing effects on intracellular calcium concentration, a possible interaction between these agents on blood pressure and heart rate was examined. With a randomized, double-blind, crossover design, 10 normal, caffeine-abstaining subjects received caffeine, 300 mg, or placebo followed by nifedipine, 10 mg, or placebo. Caffeine increased blood pressure, whereas nifedipine reduced it and caused a reflex increase in heart rate. With caffeine pretreatment, nifedipine decreased blood pressure significantly more than with placebo pretreatment. However, nifedipine reduced blood pressure to the same absolute level on both the caffeine and placebo pretreatment days. The reflex increase in heart rate after nifedipine was not affected by prior caffeine or placebo administration. Caffeine pretreatment does not alter the cardiovascular responses to nifedipine but the pressor effect of caffeine is completely reversed by subsequent nifedipine administration. PMID- 3046818 TI - Resting energy expenditure and food-induced thermogenesis in diabetic children receiving continuous subcutaneous insulin infusion. AB - The effect of short-term (8-10 days) optimal glycaemic control achieved by continuous subcutaneous insulin infusion on resting energy expenditure and food induced thermogenesis was studied. Oxygen consumption and carbon dioxide production were measured by indirect calorimetry in six newly-diagnosed and six chronically-treated diabetic children before and following the consumption of a standardized test meal. The metabolic and hormonal responses to the test meal and nutrient utilization were also assessed and compared with those measured in nine non-diabetic children. The restoration of normoglycaemia was accompanied by hypoglucagonaemia, hypoketonaemia, increased insulin:glucagon ratio and abnormal postmeal fall in free fatty acid levels in spite of normal fasting and post prandial plasma free insulin levels. These changes suggesting increased insulin action were most pronounced in newly-diagnosed diabetic children. Possibly as a result of increased insulin action high carbohydrate, low fat utilization and increased food-induced thermogenesis were observed in the newly-diagnosed diabetic children. In the chronically-treated group these parameters were approaching the normal. Resting energy expenditure was normal in both groups of diabetics. These findings suggest that precise glycaemic control can be achieved only at the expense of some degree of peripheral hyperinsulinisation which leads to altered nutrient utilization and food-induced thermogenesis in the newly diagnosed diabetic children. PMID- 3046819 TI - Somatic and autonomic nerve function during the first year after diagnosis of type 1 (insulin-dependent) diabetes. AB - Somatic and autonomic nerve function was assessed by motor and sensory nerve conduction velocities (MNCV; SNCV), beat-to-beat variation at rest, speed of pupillary dilation, and pupillary latency time in 35 newly diagnosed type 1 diabetic patients aged 12-36 years. The nerve function tests were performed 18 +/ 2 (mean +/- SEM) days after the correction of initial ketosis and hyperglycaemia and again after 3 and 12 months of insulin therapy. Mean HbA1 levels of months 3 and 12 within the normal range less than 8.6% (mean: 7.2 +/- 0.2%) were observed in 24 patients (group 1) and greater than or equal to 8.6% (mean: 10.1 +/- 0.6%) in 11 patients (group 2). Group 1 showed no significant changes from baseline in the mean nerve conduction and autonomic functions after 3 and 12 months. In group 2 there was no change until three months, however, at 12 months there was a significant decrease in mean MNCV in the median, ulnar and peroneal nerves (p less than 0.05) and in mean SNCV in the median (p less than 0.05) and sural nerves (p less than 0.01), when compared to the baseline values. The autonomic function tests remained unchanged. No patient had symptoms of neuropathy during the period studied. These findings suggest that the deterioration of motor and sensory nerve conduction precedes that of cardiac and pupillary autonomic function in poorly controlled asymptomatic type 1 diabetic patients during the first year of the disease. Effective glycaemic control prevented progression of subclinical neuropathy, but did not reverse abnormalities which were present at diagnosis. PMID- 3046821 TI - Validation of the use of 32P-orthophosphate as a probe for monitoring DNA replication in pancreatic islet cells. AB - This study aims at validating the use of 32P-orthophosphate as a probe for monitoring DNA replication instead of [3H]-thymidine where the specific activity of the latter is affected by perturbation of the cellular environment. It was found that to obtain a clean DNA fraction for determining the incorporation of the label, protein must be removed by the use of Proteinase K, and RNA by alkaline hydrolysis. Alkaline hydrolysis of RNA was found superior to RNAse digestion in not leaving behind large RNA fragments that may contaminate the DNA fraction. PMID- 3046820 TI - Changes in thermal sensation in diabetic patients after treatment with gangliosides. AB - Thirty diabetic patients (mean age (+/- SD) 45.7 +/- 11.4 years, mean duration of diabetes 7.5 +/- 7.0 years, 17 insulin-dependent), were given 3 months of placebo injections followed by either 3-4 or 6 months injections of 100 mg mixed gangliosides (Cronassial) every weekday intramuscularly in a double-blind placebo controlled study. There was an improvement of 0.77 +/- 1.25(10) degrees C and 0.65 +/- 1.05(10) degrees C in the thermal thresholds of the Cronassial treated group after 13-19 weeks and greater than 20 weeks respectively (p less than 0.22 from change of 0.04 +/- 0.59 degrees C in group treated with placebo. Compared to those with normal thresholds, patients with abnormal thermal thresholds (greater than 1.2 degrees C) improved significantly after Cronassial treatment for 13-19 weeks (-1.13 +/- 1.72(12) vs. 0.03 +/- 0.45(16) degrees C, p less than 0.006) and for greater than 20 weeks (-1.83 +/- 1.46(6) vs. 0.06 +/- 0.59(11), p less than 0.004) but this may have been related to the fact that the degree of improvement in thermal sensation was correlated with the initial thermal threshold (r = 0.80(28), p less than 0.001). There were no significant changes in electrophysiological measurements or vibration thresholds. Gangliosides are known to increase nerve sprouting and dendritogenesis and their effects in diabetic patients may be more easily seen using tests of small fibre function. PMID- 3046822 TI - Interleukin-1 and tumour necrosis factor cause hypotension in the conscious rabbit. AB - 1. The cardiovascular effects of intravenous injections of interleukin-1 (IL-1) and tumour necrosis factor (TNF) have been investigated in the conscious rabbit. They have been compared with the effects of bacterial lipopolysaccharide (LPS) because both IL-1 and TNF are released from macrophages by LPS. 2. IL-1, TNF and Escherichia coli J5-LPS all caused hypotension when given intravenously in a dose with low mortality. The time course of the hypotension caused by IL-1 and LPS was similar, although the maximal fall in mean blood pressure occurred earlier after IL-1. TNF produced a more sustained fall in blood pressure. Hypotension was not accompanied by a compensatory tachycardia after any of the test substances. Hypotension was associated with a fever after TNF, hypothermia after LPS and no significant change in temperature after IL-1. 3. The packed cell volume did not change during hypotension in any of the study groups, implying that the hypotension was not due to fluid loss resulting from increased capillary permeability. 4. IL-1 and TNF are candidates for the role of effectors of LPS induced hypotension. PMID- 3046823 TI - Longitudinal study of renal prostaglandin excretion in cirrhotic rats: relationship with the renin-aldosterone system. AB - 1. A cross-sectional study (protocol A) was performed in 19 rats with cirrhosis, induced by carbon tetrachloride (CCl4), and ascites and in 10 control animals to assess renal prostaglandin (PG) excretion in experimental cirrhosis. In an additional group of animals, including nine rats chronically exposed to CCl4 (CCl4 rats) and six control rats, a longitudinal study (protocol B) was performed to investigate the temporal relationship between changes in renal PG excretion, the renin--aldosterone system and renal function. 2. Urinary PG excretion was assessed by specific radioimmunoassay of PGE2, PGF2 alpha, 6-keto-PGF1 alpha and thromboxane (TX) B2 after extraction with octadecyl silica cartridges and h.p.l.c. purification. Recoveries for each prostanoid (61 +/- 8% for PGE2, 64 +/- 12% for PGF2 alpha, 65 +/- 11% for 6-keto-PGF1 alpha and 66 +/- 17% for TXB2) were determined in every sample by adding tritiated standards, and the final values were corrected according to the individual recoveries. 3. Cirrhotic rats with ascites in protocol A showed a significantly higher plasma renin and aldosterone concentrations and urinary excretion of 6-keto-PGF1 alpha and TXB2 than did control animals. Urinary excretion of PGE2 and PGF2 alpha, however, was significantly reduced in cirrhotic animals as compared with controls. 4. In CCl4 rats included in protocol B, there was a close chronological relationship between the activation of the renin-aldosterone system, as estimated by urinary aldosterone excretion, the onset of sodium retention and the increase in urinary excretion of 6-keto-PGF1 alpha and TXB2. The urinary excretion of PGE2 and PGF2 alpha in CCl4 rats was reduced throughout the study.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3046824 TI - Metabolic and cardiovascular effects of infusions of low doses of isoprenaline in man. AB - 1. The cardiovascular and metabolic responses to low doses of isoprenaline (15 and 5 ng min-1 kg-1 body weight infused over 30 min) were determined in six healthy males. The study was performed to investigate whether there were sustained effects after the termination of the isoprenaline infusion, as has been observed previously after the infusion of adrenaline. 2. The isoprenaline infusions produced dose-dependent increases in heart rate, systolic blood pressure and metabolic rate, but similar increases in calf blood flow and decreases in diastolic blood pressure for the two infusion rates. Finger tremor was increased in amplitude by the 15 ng min-1 kg-1 infusion only. The changes in each of these physiological variables largely resolved within a few minutes of discontinuing the isoprenaline infusions. 3. There were no changes in arterialized venous plasma adrenaline or noradrenaline levels during the isoprenaline infusions. Mean peak plasma isoprenaline levels were 0.16 +/- 0.02 nmol/l during the 5 ng min-1 kg-1 infusion and 0.71 +/- 0.05 nmol/l during the 15 ng min-1 kg-1 infusion. 4. Plasma insulin levels increased with isoprenaline but blood glucose concentrations were unchanged, consistent with a direct effect of isoprenaline on beta 2-adrenoceptors mediating insulin release from pancreatic beta-cells. Blood glycerol concentration also increased with isoprenaline but blood lactate concentration was unaltered. 5. The present study demonstrates pronounced cardiovascular and metabolic effects of low dose isoprenaline infusions. Differences in the rate of resolution of the changes induced by isoprenaline and by adrenaline seen in previous studies may result from a significant difference in their metabolism. PMID- 3046825 TI - Algodystrophy. Reflex sympathetic dystrophy syndrome. AB - Algodystrophy, which is commonly called "reflex sympathetic dystrophy syndrome" in English literature, is a common but very often unrecognized disease of the locomotor apparatus. This condition is characterized by pain with a transient, inconstant pseudo-inflammatory appearance, affecting to a variable degree the different tissue layers of one or several joint areas. Algodystrophy is often difficult to diagnose because the disease mimics the different diseases of the locomotor apparatus. The etiology, clinical symptoms, diagnosis, and treatment of this condition is discussed. PMID- 3046826 TI - Piroxicam and naproxen in patients with osteoarthritis of the hip waiting for total hip replacement. AB - Two-hundred and fifty-two patients waiting for a total hip replacement for degenerative hip disease were randomized to two groups of nonsteroidal anti inflammatory medication using piroxicam, 20 mg per day, and naproxen, 750 mg per day, after exclusion for severe dyspepsia or peptic ulcer, asthma, idiosyncracy, dissent, age below 50 years, Harris hip score above 50, or significant contralateral disease. A significant improvement in the pain and daily activity parameters was obtained in both groups. The effect was better in the piroxicam group one month after the commencement of the treatment, and equal in the groups later during the observation period of 2-5 months. We conclude that continuous medication is beneficial in patients with severe osteoarthritis scheduled for operation. However, the side effects of the medication have to be carefully considered and followed up. PMID- 3046828 TI - Dr. Douglas Taylor and half a century of musculoskeletal Kelvinism. PMID- 3046827 TI - Early alteration of synovial membrane in osteoarthrosis. AB - Biopsy specimens of the synovial membrane were obtained during arthroscopy and surgical meniscectomy from the knees of 20 patients with meniscus lesions. The aim of this study was to identify the morphological and immunological changes which appear in the synovium during the earliest phase of osteoarthrosis. Interstitial deposits of IgG and, in some cases, C3 were found not only in patients with evident arthrotic degeneration (cartilaginous lesions and synovitis), but also in patients who showed no overt arthrotic changes. Furthermore, light and electron microscopy showed an increased number of mast cells with peculiar semilunar or piecemeal aspects of their secretory granules. These ultrastructural modifications are characteristic of slow, chronic release of mediators in response to a constant, moderate degranulatory stimulus. Our findings suggest that synovial changes may occur before the advent of cartilage degeneration and that the latter may be directly influenced by early pathological changes in the synovial membrane. PMID- 3046829 TI - Acute rheumatoid factor positive (IgM) polyarthritis associated with a Klebsiella pneumonitis. AB - The authors report a case of a patient suffering from acute polyarthritis with a high rheumatoid factor titre, associated with a Klebsiella pneumonitis. A polyclonal B lymphocyte activation or a possible cross reaction between rheumatoid factor and an antigen related to Klebsiella may explain the elevated production of rheumatoid factor observed. PMID- 3046830 TI - Update on pentamidine for the treatment of Pneumocystis carinii pneumonia. AB - The chemistry, spectrum of activity, mechanism of action, pharmacokinetics, adverse effects, and dosage and administration of pentamidine are reviewed, and the role of the drug in treating Pneumocystis carinii infections in immunocompromised patients is discussed. Pentamidine isethionate, an aromatic diamidine compound, is active against certain protozoan organisms. Used extensively in the tropics in the treatment of Trypanosoma and Leishmania infections, its value in the management of Pneumocystis carinii infections has been demonstrated in infected immunosuppressed children and adults. Recently, interest in pentamidine has increased with the rising number of patients with acquired immunodeficiency syndrome (AIDS) who have P. carinii pneumonia. Pentamidine's mechanism of action and pharmacokinetic profile are not completely understood. Pentamidine is distributed extensively after i.v. or i.m. administration, with a volume of distribution of 3 L/kg. Appreciable quantities of pentamidine concentrate in the urine, and drug levels are detectable for up to six to eight weeks after cessation of therapy. After aerosol administration, the drug is almost exclusively recovered from the lung, with little extrapulmonary distribution. Available data suggest that approximately 50% of patients who receive the drug by the i.v. or i.m. route will experience some drug toxicity (local pain, sterile abscesses at the intramuscular injection site, hypoglycemia, hypotension, or azotemia), while adverse effects after aerosol therapy include bronchial irritation but little systemic toxicity. Regardless of the route of administration, pentamidine has emerged as a mainstay of therapy in the management of P. carinii pneumonitis in AIDS patients, especially in those who are allergic to the sulfa component of trimethoprim-sulfamethoxazole. PMID- 3046832 TI - Medicating the elderly. PMID- 3046831 TI - Nutritional anaemias. PMID- 3046833 TI - Primary hyperparathyroidism in the elderly. AB - Primary hyperparathyroidism is a relatively common disease in elderly women. Many of its clinical presentations may be confused with normal aging. Among the elderly, renal dysfunction and skeletal disease, particularly osteoporosis, are the two aspects of the disease likely to cause the most morbidity. It is increasingly apparent, however, that a large group of patients with hyperparathyroidism are asymptomatic or only mildly symptomatic. When symptoms are present, or repeated serum calcium levels all exceed 11.0 mg/dL, surgery is considered the treatment of choice. Success of surgery and of postoperative recovery are not affected by the age of the patient, but are enhanced by the experience of the surgeon performing the parathyroidectomy. There are difficult management decisions to be made concerning the elderly asymptomatic patient with mild hypercalcemia. A conservative approach to therapy for these individuals would include maintenance of adequate hydration, and involvement in physical activity. Dietary restriction of calcium should be recommended only as long as there is no evidence that it is exacerbating negative calcium balance. Drugs likely to worsen hypercalcemia, such as thiazides, must be avoided, and blood chemistries should be monitored at regular intervals. Because the bone loss of hyperparathyroidism may develop insidiously, serial bone density measurements are probably reasonable in older women already at risk for osteoporosis. Accelerated bone loss is an indication for surgical intervention. A wide variety of medications have been proposed for the treatment of primary hyperparathyroidism, but none stands out as a particularly desirable therapeutic alternative. Future research in this area, particularly with the use of estrogens in postmenopausal women with hyperparathyroidism, may eventually lead to greater acceptance of this alternative to surgery. PMID- 3046835 TI - Nonpharmacologic treatment of age-associated memory impairment. PMID- 3046834 TI - Management of non-inflammatory musculoskeletal disorders in the elderly. PMID- 3046836 TI - Lung cancer in the elderly. PMID- 3046837 TI - Anemia in the aged. PMID- 3046838 TI - Life span extension and food restriction. PMID- 3046839 TI - Ethics in medical research: Australian developments. PMID- 3046840 TI - Repetition strain injury and psychiatry. PMID- 3046841 TI - Psychological stress as a public health problem: how much do we know? PMID- 3046842 TI - Class inequalities and 'just health' strategy. PMID- 3046844 TI - MHC class I gene expression by tumors: immunotherapeutic implications. PMID- 3046843 TI - Immunogold-EM localization of actin and vimentin filaments in relation to polygonal arrays in lens epithelium in situ. AB - An indirect immunogold technique for transmission electron microscopy was used for localizing two cytoskeletal proteins, actin and vimentin, in the epithelium of freshly removed rabbit lens, especially in relation to the polygonal array structures located at the apices of the epithelial cells. Antibody specificity was determined on semi-pure chicken breast muscle actin and bovine lens vimentin using Western blotting of these proteins and extracts of rabbit lens epithelium separated by SDS-PAGE. Whole lenses of rabbits were lightly fixed in glutaraldehyde and embedded in LR White resin. Tangential sections were taken at 70 to 80 nm and at 0.25 micron and used for single-labeling, and double-labeling with antibodies raised in different hosts and treated with appropriate second antibodies conjugated with non-overlapping sizes of gold particles. Routine and stereomicroscopy were used to analyze gold-label patterns. The study shows that the rays of the polygons project deeply into the cell from the vertices lying on the inner apical membrane. Actin is located on the filaments of rays, but vimentin is not associated with the polygons at the level in the cell that we studied. Vimentin filaments are found in deeper regions of the epithelial cell. Stereopairs were useful in differentiating where the gold-label was located and in fact, this technique demonstrated that most of the label is on the surface of sections where the filaments are exposed. PMID- 3046845 TI - Subungual trichogranuloma in a hairdresser. AB - Hairdressers may be subject to occupational skin diseases other than contact dermatitis. Interdigital trichogranuloma is a common disorder among those who cut men's hair but is much less common among those who cut women's hair. Interdigital trichogranulomas or interdigital sinuses are the result of penetration of the skin by short, sharp hair clippings. Subungual hair penetration appears to be much less common. This article reports the first case of subungual trichogranuloma in a hairdresser with psoriatic onycholysis. Onycholysis may be both a risk factor for and a consequence of cut hairs becoming imbedded subungually. PMID- 3046846 TI - The creeping vole, Microtus oregoni: karyotype and sex-chromosome differences between two geographical populations. AB - The G-banded karyotype of the creeping vole, Microtus oregoni, prepared from animals trapped in Oregon and Washington, is presented. The two populations had similar autosomal banding patterns but exhibited striking differences in their sex chromosomes. The X chromosome of voles captured in Oregon was 39% longer than that of voles trapped in Washington. The length difference was primarily due to an increase in size of light G-bands, which, in both populations, comprised large segments of the X chromosome. On C-banding, the X chromosome exhibited major blocks of constitutive heterochromatin corresponding to the light G-bands. In contrast, the Y chromosome of the Oregon voles was 24% shorter than that of the Washington voles and lacked the short arm and some terminal bands present in the Washington voles. PMID- 3046848 TI - [Clovis Vincent 1879-1947]. PMID- 3046847 TI - [Etiology of congenital dilatation of the choledocus in children]. PMID- 3046849 TI - [Awarding the International Lannelongue Medal to Dr Norman E. Shumway]. PMID- 3046850 TI - [Intestinal cystic pneumatosis. Apropos of 2 cases]. PMID- 3046851 TI - Antimicrobial agents as biological response modifiers (BRM) and chrono immunomodulation: an emerging relationship. AB - Immune defense mechanisms play an essential protective role against infections caused by a wide array of pathogenic microorganisms. Although the growing number of the available antimicrobial agents has certainly improved the overall clinical outcome of such infections, antimicrobial therapy not rarely fails whenever the host's immune function is depressed. On the other hand, recently introduced therapeutic and diagnostic procedures (antineoplastic chemotherapy causing severe neutropenia and mucositis; organ transplantation requiring conditioning regimens; the widespread use of intravascular catheters and prosthetic devices; administration of adrenal corticosteroids and/or other immunosuppressive agents) have resulted in an unprecedented number of immunocompromised hosts. In addition, a variety of antibiotics have been found to display adverse effects on specific and non-specific immune functions, thus further impairing the already depressed immune system of the host. Antibiotic-mediated immunomodulation hence is explored with the introduction of a third-generation cephalosporin, namely cefodizime (CDZ), which has been shown to possess immunostimulating properties in preliminary in vitro and ex vivo studies as well as in a few experimental animal models. A chronoimmunopharmacological approach to CDZ-induced immunomodulation has been started by ourselves. The study, which is still in progress, includes patients with multiple myeloma (MM), selective IgA deficiency and chronic uremia, and matched healthy subjects. A number of immunological parameters are being assessed on blood samples drawn every 6h in the 24-h span prior to CDZ administration (a single 2 g daily dose i.v. for 6 days to the patients and for 4 days to healthy subjects), and in the 24h following the last CDZ injection. Healthy subjects and patients are randomly assigned to two groups, depending on whether they are given the antibiotic at 0800 or at 1800. Although a full evaluation of the results will be reported elsewhere, the group of MM now includes 24 patients. A circadian stage-dependent chronoimmunomodulating effect has been unequivocally shown for the monocytic chemotactic responsiveness to CDZ in MM. Immunostimulating 'side-effects' suggest that CDZ should possibly be regarded as a prototype antimicrobial agent for patients with impaired immune functions. Conceivably, a better knowledge of such properties will help synthesize new antibiotics with specific immunomodulating effects. PMID- 3046852 TI - Multifrequency rhythms of immunological functions. AB - Chrono-immunological functions are reviewed starting from the first studies on circadian variations in the count of circulating blood eosinophils to the most recent studies on lymphocyte subpopulation periodicities. Serum Ig levels, complement components, plaque forming cells, phagocytosis, autologous mixed lymphocyte reactions, have also been demonstrated to be circadian periodic. Transplant immunology is also known to show circadian and circa septan periodicities in the rejection of transplanted organs. Rhythms of susceptibility to a series of allergens and bronchial asthma symptoms characterize chronobiology in allergy. A periodic pattern is a common finding in the recurrence of the symptoms in rheumatoid arthritis. Finally, the beginnings of chrono-immunotherapy of experimental and human tumors are briefly reviewed. The conclusion that can be drawn from this short overview is that the demonstration of the periodicity in such a big number of variables in health and the modifications of the relevant parameters in disease, emphasize the importance of the predictable, since periodic, part of the variations that can be shown by the immunological findings. PMID- 3046853 TI - Effect of fat-free diet on insulin requirements in type I diabetes controlled with artificial beta-cell. AB - We investigated the effect of eliminating calories derived from fat sources on postprandial and basal insulin requirements in five patients with type I (insulin dependent) diabetes mellitus. The patients were studied on a metabolic ward on two solid-food diets with similar quantities of carbohydrate and protein with or without the addition of fat. Diet A was isocaloric (weight maintenance) with calories distributed as 45% carbohydrate, 15% protein, and 40% fat. Diet B contained the same carbohydrate and protein content as diet A but was virtually fat free and therefore hypocaloric (1233 +/- 106 vs. 1830 +/- 99 cal, mean +/- SE). The diets were given as five equal meals each day on consecutive days. Insulin requirements and blood glucose measurements were determined by use of the artificial beta-cell. During the study, mean (+/- SE) preprandial blood glucose levels were maintained at 85 +/- 11 mg/dl, and peak postprandial blood glucose levels were less than 180 mg/dl. The elimination of fat calories had no effect on total (68.9 +/- 10.3 vs. 69.3 +/- 4.9 U/day), postprandial (9.8 +/- 3.8 vs. 10.3 +/- 3.7 U/meal), or basal (1.9 +/- 0.2 vs. 1.8 +/- 0.2 U/h) insulin requirements. Thus, despite a hypocaloric diet, no change in insulin requirements was noted when fat-derived calories were deleted from the diet. We conclude that fat derived calories do not alter short-term basal or postprandial insulin requirements in type I diabetes. PMID- 3046854 TI - Response to single dose of aspartame or saccharin by NIDDM patients. AB - Twelve normal subjects and 10 subjects with non-insulin-dependent diabetes mellitus were given, in random order at intervals of greater than or equal to 1 wk, three drinks of the same beverage: one unsweetened, one sweetened with 400 mg aspartame, and one sweetened with 135 mg saccharin. The amount of sweetener approximated that in 1 L of sugar-free soft drink. Plasma glucose, insulin, and glucagon were measured for 3 h after ingestion of the test beverage. Plasma glucose declined slightly throughout the test period, probably due to fasting, with no differences between the three treatments. Neither sweetener affected peak insulin levels in subjects with or without diabetes. Analysis of area under the curve showed that mean insulin levels were statistically significantly higher after aspartame than after saccharin or unsweetened beverage in normal subjects only, but the magnitude of the difference was small and unlikely to be of physiological importance in the absence of differences in glucose levels. Furthermore, the differences could largely be accounted for by a decrease in insulin values after both unsweetened beverage and saccharin, with no change from baseline after aspartame. Glucagon levels showed time-to-time variation but no overall differences. We conclude that ingestion of aspartame- or saccharin sweetened beverages by fasting subjects, with or without diabetes, did not affect blood glucose homeostasis. PMID- 3046856 TI - Insulin levels after injection by jet stream and disposable syringe. PMID- 3046855 TI - Cyclosporin A in treatment of new-onset type I diabetes mellitus. PMID- 3046857 TI - History of ophthalmology, 1. PMID- 3046858 TI - The first German textbook of ophthalmology "Augendienst" by G. Bartisch, 1583. AB - This book contains various illustrations, portraits and an exact index, testimonials proving the author's professional successes as well as an accurate list of the qualities that should be demanded from any ophthalmologist. The anatomy of the head and eye is described according to Galen's ideas and Vesalius' book. Many remedies, prescriptions and medical treatments are discussed, partly showing the mystic influences of the Middle Ages. Bartisch reports several diseases for the first time: Allergic reactions, sympathetic ophthalmia, hemeralopia, photoelectric keratoconjunctivitis, amaurosis due to toxemia of pregnancy. But most important is the part on surgery. A careful pre- and postoperative treatment is demanded in cases of cataract operations. Bartisch describes the removal of eyelashes to cure trichiasis, the operations of ptosis, blepharochalasis and the exenteration of the orbit. This book was appreciated for a long time so that in 1686 a nearly identical reprint was published. PMID- 3046859 TI - The professors of ophthalmology at the University of Leipzig in the first half of the 20th century. PMID- 3046860 TI - "Eye injuries" by the Byzantine writer Aetios Amidinos. AB - The authors give a short report of the "Injuries of the Eye" by the Byzantine writer Aetios Amidinos. He wrote 16 medical books, among them "The 7th Logos", including Eye Diseases. In all his writings, there is a substantial resort to ancient medical literature, mainly to the books of Galen. He describes the Eye Injuries in 5 chapters with remarkable critical ability and suitable treatment. Thus he is acknowledged as one of the most cultured and productive writers of that period. PMID- 3046861 TI - The lacrimal surgery of Petrus Camper and his contemporaries. AB - Lacrimal surgery went through a period of rapid development in the 18th century. This is all the more remarkable because at that time anaesthesia was still unknown. In his ophthalmological textbook "De Oculorum Fabrica et Morbis", written in 1766, the renowned Dutch anatomist and surgeon Petrus Camper (1722 1789) presented a detailed review of the various operative procedures performed by himself and his contemporaries. PMID- 3046862 TI - The French Egyptian campaign and its effects on ophthalmology. AB - Almost all soldiers of the armies involved in the Egyptian campaign fell victim to what was later called the ophthalmia militaris which we now know to be caused by Haemophilus aegyptius, N. gonorrhoea and possibly to some extent by Chlamydia trachomatis but more likely by the adenoviruses. Because of the enormous incidence of ocular infection and the controversy generated by speculation on the nature of the disease--English surgeons considered this ophthalmia to be of a contagious nature, whereas the French surgeons violently opposed this view-, the interest in diseases of the eye increased, which eventually resulted in the acceptance of ophthalmology as a separate branch of medicine. PMID- 3046863 TI - The appointment of Johan Widmark to the first chair in ophthalmology in Stockholm 1891. PMID- 3046864 TI - A historical outline of Greek ophthalmology from the Hellenistic period up to the establishment of the first universities. AB - The writers examine the course of Greek ophthalmology from the Hellenistic period to the foundation of the first universities (19th century). In particular, the study refers to Galen, Antyllus, the Byzantine doctors Oribasius, Aetius of Ameda, Paul of Aegina, Alexander of Tralles, Nonnus Theophanes, Theophilus Protospatharius, Michael Psellos, Meletius Monachus, Nemesius bishop of Emeses and John Actuarius. The practice of empirical ophthalmology during the Ottoman domination of Greece is also examined, as is the earliest available evidence of modern Greek ophthalmological knowledge, deriving from the Ionian Islands. PMID- 3046865 TI - History of the Japanese Ophthalmological Society. PMID- 3046866 TI - The general development of Chinese ophthalmology from its beginnings to the 18th century. PMID- 3046867 TI - Leonardo and the eye. PMID- 3046868 TI - Four legends about Hippocrates. Collected by Mr. Zarakas of the Village Pili-Cos. PMID- 3046870 TI - The representation of the eye in African and Oceanian art. PMID- 3046871 TI - The Utrecht Ophthalmic Hospital and the development of tonometry in the 19th century. AB - During the second half of the 19th century Donders, Snellen and co-workers of the Utrecht Eye Clinic played an important role in the development of clinical tonometry. These indefatigable researchers designed and built a number of tonometers of which most have been saved and which are now on display in a permanent exhibition in the Royal Netherlands Ophthalmic Hospital at Utrecht. PMID- 3046869 TI - Ophthalmological lore in the Corpus Hippocraticum. AB - In this paper we examine the ophthalmological knowledge of the Hippocratic School as described in the Corpus Hippocraticum. An analysis is made of knowledge existing at the time concerning the anatomy of the eye and the physiology of vision, as well a diseases of the eye and ophthalmic surgical operations as depicted in the Corpus. In particular, the book "About Vision" from the Corpus Hippocraticum is discussed in detail. From our study of the extant texts it becomes apparent that not only was knowledge of ophthalmology considerably developed in the time of Hippocrates but it constituted a source of inspiration for ophthalmic treatments carried out by later physicians and in particular by those of the 19th century. PMID- 3046872 TI - The Utrecht Ophthalmic Hospital and the development of the ophthalmoscope. AB - The first useful table model ophthalmoscope was designed by Donders and the instrumentmaker Epkens. With the use of this instrument Donders' pupil Van Trigt made the first drawings of the normal and pathological fundus of the eye. His contribution was of great importance for the early diffusion of knowledge of the fundus of the eye. PMID- 3046873 TI - The foundation of experimental ophthalmology by Theodor Leber. AB - Theodor Leber grew up in Heidelberg as the son of a professor of Romance languages. Initially he planned to study natural sciences. Bunsen's advice led him to medicine. During his studies he succeeded in solving a competition problem posed by Helmholtz in the medical department. A short period of practical work in the eye hospital of Knapp was unsatisfactory. In Vienna with the physiologist Carl Ludwig, he was able in 1863/64, at the age of only 24 years, to demonstrate the blood circulation of the eye by color injections into the arteries and veins. Since that time the schematic drawings of his results can be found in every textbook of ophthalmology. On the occasion of the congress of the German Ophthalmological Society in Heidelberg in 1864, Theodor Leber reported on these findings and met with immense approval. In 1864-67 he followed an invitation as coworker of Liebreich to Paris; in 1867 he became A.v. Graefe's coworker in Berlin; in 1871 he moved to Gottingen, which became the first eye clinic with a laboratory for experimental investigations. The second epoch-making discovery accomplished by Leber was the detection of the fluid exchange in the eye. These results have also been confirmed by modern methods. Therefore, Theodor Leber can be called the father of experimental ophthalmology. PMID- 3046875 TI - Eye votives in Greek antiquity. PMID- 3046874 TI - Georg Joseph Beer: a review of his life and contributions. PMID- 3046876 TI - [Computer diagnosis of gastric dysplasia and program design]. AB - Two step processes were taken in computer diagnosis of gastric dysplasia. 1. 5 measurement values of dysplastic glandular tube obtained by using image analysis system were taken as the basis for judging the degree of dysplasia. Three glandular tubes of dysplasia were measured in each case. 2. Computer pathology diagnostic program using BASIC language ran in IBM computer. 5 parameters were input into computer for three stage-diagnosis. The first stage: 5 parameters were counted basing on three models to obtain three values which reflected the objective feature of the same glandular tube from different aspects. The second stage: diagnosis of a dysplastic glandular tube was obtained depending on the three values judged synthetically. Variate 'N' was introduced for controlling the first and second stage diagnosis of the three glandular tubes. The third stage diagnosis started as soon as N = 3. The third stage: diagnostic values of three glandular tubes were balanced and analysed depending on diagnostic criteria to reach the final diagnosis. PMID- 3046877 TI - [Luteinizing hormone-releasing hormone (LRH) agonist in the treatment of breast cancer, osteosarcoma and prostate carcinoma--analysis of 12 patients]. AB - This paper first reports the results of 12 patients with sex hormone-dependent neoplasms, including 9 women with breast cancer, 2 men with osteosarcoma and 1 man with prostate carcinoma, treated by LRH agonist, (D-Ala6, des-Gly-NH2(10)) LRH-ethylamide (LRH-A) 100-200 micrograms, IM, QD. After 15-30 days of administration, the concentrations of plasma mean estradiol, progesterone, testosterone, serum luteinizing hormone and follicle-stimulating hormone were lowered significantly in the peripheral blood of all patients, associating with improvement of the patients' general condition and reduction of the tumors and/or metastatic foci. No serious side effects were observed except vaginal irregular bleeding in isolated patients. Three patients died and the others were alive in follow-up of 4-30 months. The results suggest that LRH-A be useful in the treatment of the sex hormone-dependent tumors and worth further study. PMID- 3046878 TI - Sulfonylurea-induced decrease of muscle capillary basement membrane thickness in diabetes. AB - Three muscle biopsies were performed in 53 overt type 2 diabetics over a period of approximately 2 years. At baseline, 21 (40%) had an increased capillary basement membrane width in muscle. Thirty-five patients received glipizide and 18 received placebo. In the patients receiving placebo, the mean of the muscle capillary basement membrane width increased from 158.7 +/- 11.5 nm (SEM) to 170.9 +/- 14.7 nm (P = NS), but in those receiving glipizide the value decreased from 192.9 +/- 13.2 nm to 161.0 +/- 10.2 nm (P = 0.02). Plasma glucose and glycosylated hemoglobin A1 decreased significantly (P less than 0.001) after 2 years in patients receiving glipizide. In 15, mean glycosylated hemoglobin A1 reached a normal range, and mean basement membrane width decreased to a level close to that found in subjects without diabetes (P = NS). These findings are consistent with the hypothesis that effective response to oral medication can decrease the basement membrane thickening, suggesting that diabetic microangiopathy is not necessarily progressive. PMID- 3046879 TI - Ultrastructural and functional studies of pancreatic B cells in Wistar rats treated with immunotherapeutic doses of cyclosporin. AB - Cyclosporin (CYA) was administered to Wistar rats at immunotherapeutic doses (20 mg/kg) and the functional and morphological effects on pancreatic B cells were observed. Serum CYA levels were kept in the range that is often encountered in clinical immunotherapy in humans. The treated rats had slightly elevated plasma glucose levels; on treatment days 6 and 13, their serum immunoreactive insulin and pancreatic insulin contents were low. Plasma immunoreactive glucagon levels were slightly elevated during treatment. Glucose tolerance was impaired on day 13. These functional abnormalities disappeared 2 weeks after drug withdrawal. B cells showed ultrastructural evidence of CYA toxicity, including cytoplasmic degranulation, nuclear inclusions, and cisternal dilatation of both the rough endoplasmic reticulum and the Golgi apparatus. These changes were observed on the third day of treatment and became more pronounced on the 6th and 13th days of treatment. By the second week after drug withdrawal, most of the B cells presented a normal ultrastructure. We confirmed that a dose of CYA similar to that used in clinical immunotherapy in humans produces ultrastructural disturbances in the pancreatic B cells of Wistar rats after relatively short-term administration. PMID- 3046880 TI - Different effects of growth hormone on growth and function of isolated adult and fetal rat pancreatic islets. AB - We simultaneously determined 3H-thymidine incorporation, DNA content, and insulin biosynthesis in adult and fetal rat pancreatic islets. 3H-thymidine incorporation was measured after tissue solubilization. DNA was measured fluorometrically. Insulin biosynthesis was determined through 3H-leucine incorporation after immunoprecipitation and binding to protein A-sepharose. Addition of growth hormone caused a significant increase in 3H-thymidine incorporation into both the adult and the fetal islets, but only the adult islets experienced an increase in DNA content. The increase in 3H-leucine incorporation was also significant only in the adult islets. The dynamics of 3H-thymidine incorporation after growth hormone administration were different in the adult and the fetal pancreas. Fetal islets have the potential to increase their incorporation of 3H-thymidine, but very limited potential for insulin biosynthesis. PMID- 3046881 TI - [Topography of the mRNA-binding region of the ribosomal protein S1 from Escherichia coli]. PMID- 3046882 TI - The role of histamine in the cardiovascular system. AB - This article reviews briefly the role of histamine through its H1 and H2 receptors on the cardiovascular system and its action on calcium and catecholamines. The analogy between the adrenergic and the histaminergic systems is well demonstrated and there is evidence that histamine participates in myocardial damage and arrythmias, but the question of its exact role in the early stages of cardiovascular diseases, such as myocardial ischaemia and atherosclerosis, requires further study. PMID- 3046884 TI - Efficacy and tolerance of cicletanine, a new antihypertensive agent: overview of 1226 treated patients. AB - The data gathered from 1226 patients for 20 controlled therapeutic studies performed during the phase-III development of cicletanine, a new antihypertensive agent, were pooled in a computerized database. An extensive statistical analysis of these data collected over 1-2 years was performed to give a clear interpretation of long-term efficacy and tolerance of this drug. In mild to moderate hypertension, the dose-response relationship observed after one month of treatment on diastolic (DBP) and systolic (SBP) blood pressure disappeared during the third month. After a 3-month treatment in monotherapy with the recommended dosage (50-100 mg/day) we observed a mean decrease of 29.7 +/- 17.8 mmHg for SBP and 23.3 +/- 13.2 mmHg for DBP. At this stage 70.9% of patients were stabilized (SBP less than 160 mmHg and DBP less than 95 mmHg) with cicletanine. After this initial regular decrease of blood pressure values, an additional decrease of several mmHg was observed during the 24 months of treatment, thus significantly augmenting the number of stabilized patients. No significant difference in efficacy was observed in adult and elderly (65 to 95 years) patients. The clinical tolerance was very good, only a few slight and transitory side-effects have been reported. Biological tolerance was also very good. Depending on the dosage used, only a slight and transitory variation in sodium and potassium levels was observed. Glucose, creatinine and lipids either remained stable or were improved during treatment. This analysis demonstrates the beneficial effect and the good tolerance of cicletanine (50-100 mg/day) in the treatment of hypertension, which can be explained by its special mode of action. PMID- 3046883 TI - Review of three studies to determine the efficacy and tolerance of cicletanine in the short- and long-term treatment of essential hypertension. AB - A double-blind multicentre study was performed initially to assess the effect of 4 weeks' treatment of cicletanine in daily doses of 12.5, 25, 50 and 100 mg, for the treatment of 60 patients with mild to moderate essential hypertension. A significant reduction in blood pressure was achieved with each dosage. Analysis of the responder rate indicated a significant dose-response relationship which was clinically relevant only for the 50 and 100 mg doses. The minimum therapeutic dosage of cicletanine for the treatment of essential hypertension was concluded to be 50 mg daily. A further 12-week study was performed in 40 patients with mild to moderate essential hypertension, commencing with a daily dose of cicletanine (50 mg) and increasing the dosage if necessary to 100 mg and 200 mg daily after 4 and 8 weeks, respectively. Blood pressure control was achieved in all patients and in the majority (70%) with a daily dosage of 100 mg cicletanine. Treatment was continued in all patients until a total duration of 2 years had been completed. Cicletanine was shown to be very well tolerated and a significant reduction in blood pressure was achieved at 6, 9, 12, 18 and 24 months. During the second year of treatment, all the patients were stabilized with a daily dosage of 50 mg cicletanine. These results are consistent with a previously reported study in 25 patients with mild to moderate essential hypertension. Twelve weeks of treatment with cicletanine (50 mg daily) produced a significantly greater reduction in blood pressure than a combination of 5 mg amiloride hydrochloride and 50 mg hydrochlorothiazide daily. This reduction was more significant for the diastolic than the systolic blood pressure measurements. A diastolic blood pressure of 90 mmHg, or less, was achieved in all patients receiving treatment with cicletanine (50 mg daily), except for two patients where the dosage was increased to 100 mg daily. PMID- 3046885 TI - Study of the effects of cicletanine on the renin-angiotensin-aldosterone system. AB - The effect of a single dose of 50 mg of cicletanine on plasma renin activity, plasma potassium and aldosterone, blood pressure, urinary volume and sodium and potassium excretion was compared to the effect of the same dose given for 9 days in eight normal volunteers in metabolic balance on 120-140 mEq of sodium and 50 70 mEq potassium. Following 4 days of this standardized diet, the normal volunteers received their first dose of cicletanine after an overnight fast and blood was collected at 30 min, 2, 4 and 6 h for plasma renin activity and aldosterone. Urine excreted following a modest water load was also collected and analysed over a 6-h period. Following the single dose there was an increase in PRA at 1 and 2 h, while there was no such increase with chronic administration. Plasma aldosterone, however, showed no difference between acute and chronic administration of cicletanine treatment. Also plasma uric acid levels increased with acute but not chronic cicletanine. There were no differences in heart rate and blood pressure responses, nor in urine volume, urinary sodium or potassium excretion, or plasma potassium between the different treatments. This study strongly suggests that the antihypertensive effect of cicletanine is not the result of an increased diuresis, as in this case enhanced urinary electrolyte excretion and increased plasma renin activity and aldosterone would be mandatory following chronic administration. Recent evidence suggests that the dissociation between plasma renin activity and aldosterone is the consequence of an effect of cicletanine on intracellular calcium mobilization. This mode of action would explain both the antihypertensive as well as the aldosterone secretion reducing effect of cicletanine. PMID- 3046886 TI - Urinary incontinence in the elderly: etiology and treatment. AB - Urinary incontinence is a common, though often hidden, medical problem among the elderly. Urinary continence requires integrity of the neural, muscular, and hormonal systems. Five distinct types of urinary incontinence can be distinguished based on patient symptoms. A variety of factors can impair continence, including aging, environmental barriers, and medications. Both pharmacological and nonpharmacological measures are useful in the treatment of incontinence. PMID- 3046887 TI - Sulbactam/ampicillin, a new beta-lactamase inhibitor/beta-lactam antibiotic combination. AB - Sulbactam/ampicillin is a combination of a beta-lactamase inhibitor with minimal intrinsic antibacterial activity (sulbactam sodium), and an aminopenicillin (ampicillin sodium). The addition of sulbactam to ampicillin has no effect on the chemical stability of ampicillin in aqueous solution, and the administration guidelines of the combination are the same as for ampicillin alone. Sulbactam acts primarily by irreversible inactivation of beta-lactamases from most beta lactamase-producing organisms. The pharmacokinetics of sulbactam are similar to those of ampicillin with an elimination half-life of about one hour in most patients. One difference is that serum and tissue concentrations of sulbactam are usually twice those of ampicillin, at equivalent doses. The sulbactam/ampicillin combination has been approved for the treatment of adults with intraabdominal, skin and skin structure, and gynecological infections due to beta-lactamase producing bacteria such as Staphylococcus aureus, Escherichia coli, and species of Klebsiella and Bacteroides. Clinical studies to date have also shown the combination to be effective for the treatment of meningitis, pneumonia, gonorrhea, epiglottis, urinary tract infections, cervical adenitis, and as prophylaxis for abdominal and gynecological surgeries. Many of these studies, however, have included small numbers of patients and/or had design flaws. Adverse effects have been minor with most being attributed to the ampicillin component. Sulbactam/ampicillin compares favorably with other antibiotic regimens in terms of acquisition costs and ease of administration. PMID- 3046888 TI - Lovastatin for hypercholesterolemia. AB - Lovastatin is the first 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitor approved for the treatment of primary hypercholesterolemia. It is indicated as adjunctive therapy to dietary control and should be initiated at 20 mg/d in the evening. With higher dosages, twice-daily dosing is preferred, particularly when the dosage reaches the maximum recommended 80 mg/d. Compared with other drugs available, lovastatin has been shown to have good efficacy and a low incidence of side effects. Limited pharmacokinetic information available from the manufacturer reports absorption approximately 30 percent, protein binding greater than 95 percent, and a dual pathway for elimination through both urine (10 percent) and feces (83 percent). The drug has been clinically tested versus placebo and in combination with other cholesterol-lowering drugs. Lovastatin is effective in lowering total cholesterol and low-density lipoprotein cholesterol by 25-30 percent, with nonfamilial (hypercholesterolemic) patients responding better than those with the familial form of the disease. One percent of lovastatin patients have discontinued therapy because of intolerable side effects. The most common complaints are flatulence and diarrhea; more severe abnormalities include elevation of liver enzymes and an unclear propensity for producing lens opacities. The monthly cost to a patient taking 20 mg/d is approximately $44. Although the drug should be added to hospital formularies, long-term safety experience and competition from other HMG-CoA reductase inhibitors will determine lovastatin's final therapeutic role. PMID- 3046889 TI - Levothyroxine sodium tablets: chemical equivalence and bioequivalence. AB - Two brands of levothyroxine sodium tablets were compared in vivo for bioequivalence in a double-blind, randomized study. The tablets were identical in levothyroxine content. Evaluation was by means of triiodothyronine (uptake), tetraiodothyronine, free thyroxine index, total triiodothyronine by radioimmunoassay, and thyroid-stimulating hormone measurements. No differences in clinical response were found in a study with a high statistical power. It was concluded that the two brands were bioequivalent because of chemical equivalence, use of micronized levothyroxine powder in tablet production with at least one of the brands, and scrupulous attention to quality control during the manufacturing process, all of which contributed to assurance of homogeneity of the products and close adherence (+/- five percent) to the claimed potency. PMID- 3046890 TI - Carbocysteine. AB - Carbocysteine is prescribed for conditions characterized by the accumulation of excessive mucus. Although often described as a mucolytic, its function is probably that of mucoregulation, which results in physical changes in accumulated secretions that are favorable in terms of clearance. The chemistry, pharmacology, pharmacokinetics, clinical applications, and toxicology of carbocysteine are reviewed. PMID- 3046891 TI - Calcium-channel blocking agents and chest pain. AB - Calcium-channel blocking agents can relieve anginal and chest pain associated with hypermotility of the esophagus. Although calcium-channel antagonists differ in their effects on the esophageal body and the lower esophageal sphincter, reduction of lower esophageal sphincter pressure theoretically could result in gastroesophageal reflux and pain. This pain may be difficult to differentiate from chest pain of cardiac origin. An association between calcium-channel blocking agents and chest pain is speculative at this time. However, the possibility of such an effect should be considered by clinicians in the total management of patients. Future clinical studies are warranted to address the issue of the incidence of such an effect and to define more precisely the potential interrelationship among the calcium-channel antagonists, their efficacy in relieving cardiac pain, and their potential liability in inducing chest pain of noncardiac origin. PMID- 3046893 TI - [Shockwave lithotripsy of gallstones]. PMID- 3046892 TI - Cost comparison of ceftazidime versus tobramycin/ticarcillin therapy in three hospitals. AB - A retrospective review of 114 patient charts was conducted at three geographically different hospitals to compare the costs of ceftazidime (CTZ) with the combination of tobramycin and ticarcillin (T/T) in the treatment of hospital acquired pneumonias and/or septicemias. Variables analyzed to determine the cost of the therapy included duration of treatment, dosage and number of doses administered, side effects, and laboratory tests. Time and motion studies were also performed and the results included in the algorithm. In two of the three hospitals, the computer-calculated cost of CTZ therapy was significantly less than for T/T therapy ($1194 vs. $1668 and $93 vs $629). In the third hospital there was no significant difference in the costs of the two treatments ($1079 for CTZ vs. $1066 for T/T). The cost of each regimen per patient day followed a similar pattern, with CTZ therapy being significantly less expensive in the same two hospitals. Patients receiving CTZ therapy realized an average cost savings of $29.70 per day at the three institutions. These savings appeared to be due to a reduction in laboratory and administration costs resulting from the decreased frequency of CTZ administration. We conclude that CTZ therapy was less costly than T/T therapy at three geographically different institutions when the cost of supplies, laboratory tests, and personnel time were taken into account. PMID- 3046894 TI - [Significance of ultrasonics in the placement of a central venous catheter]. AB - An ultrasound investigation was undertaken of the neck region of 42 patients with normal neck anatomy in order to determine whether the results of ultrasound gained topographical data provided pointers to the choice of entry site to the internal jugular vein (IJV). In addition, the IJV was punctured under ultrasound control in 23 patients in an intensive care unit in whom there was a problem of increased bleeding tendency, anatomical difficulty or previously failed "blind" puncture. In all of them a central venous catheter was placed without complication by the Seldinger technique via the primary chosen point for puncture. An approach through the sternocleidomastoid muscle, between the cricoid level and the "central" place of puncture between the two bellies of the sternocleidomastoid muscle proved to be the most satisfactory compromise between easy application of the ultrasound head, large vein diameter and reduction of any risk of mistakenly puncturing artery or pleura. This approach has to be varied according to the ultrasound findings. It is concluded from this experience that ultrasound is suitable for the placement of central venous catheters. But since the equipment is bulky it cannot be used in an emergency. PMID- 3046896 TI - [Chylous lymphocele: a rare complication after gastrectomy for primary non Hodgkin's lymphoma of the stomach]. AB - Gastrectomy and extensive lymphatic resection were performed in a 55-year-old woman with primary non-Hodgkin lymphoma of the stomach. Three months later a large perihepatic lymphocele developed. After placement of a catheter under ultrasound control and evacuation of about 5.31 of chylous fluid there was no recurrence or further sequelae. This complication is so rare that it should not be a reason for limited lymphatic resection in malignant disease. PMID- 3046897 TI - [Long-term intragastric pH-metry. Methods and clinical significance]. PMID- 3046895 TI - [Immunodiagnostic findings in patients with kala-azar]. AB - Using three immunoreactions (complement-fixation reaction; enzyme immune test; indirect immunofluorescence), antibody formation was tested in 66 patients with kala-azar and 74 controls (blood donors; accident patients). Moderately elevated and high antibody levels for each of three reactions were defined to provide diagnostic criteria. Moderately high or high antibody concentrations against Leishmania antigen were found in one of the three immunoreactions in 9 patients, in two of the three in 21, and in all three in 36 (55%). No Leishmania antigen was found in the serum of the controls. Similar results were obtained for 80% of those serum samples which had been sent in over a period of seven months to check for Leishmania antibodies (455 of 566 samples). In 25 of the remaining 111 serum samples moderate or high antigen concentrations were demonstrated, but in 21 of them in only one test. Leishmania antibodies were found to persist for several months after the clinical symptoms had disappeared. In 22 kala-azar patients the IgG concentrations were clearly elevated (greater than 3000 mg/dl), in five the IgM concentrations were elevated (greater than 400 mg/dl), while the IgA concentrations were normal or slightly decreased. The data indicate that kala azar cases can be diagnosed with three appropriate immunoreactions which measure antibody concentrations. PMID- 3046898 TI - [Pathogenetic importance of eosinophilic granulocytes. II. Hypereosinophilic diseases]. PMID- 3046900 TI - [16th Environmental Health Seminar: "Hygienic aspects of pork production". WHO Collaborating Centre for Research and Training in Veterinary Public Health and the Hannover School of Veterinary Medicine. Proceedings]. PMID- 3046899 TI - [Objective ascertainable indicators of pork quality]. PMID- 3046901 TI - [The influence of breeding precautions on the meat quality in swine]. PMID- 3046902 TI - [The influence of feeding on the meat quality in swine]. PMID- 3046903 TI - [The influence of housing arrangements and production hygiene on meat quality]. PMID- 3046904 TI - [The significance of the use of veterinary drugs in swine production and the effects on meat quality]. PMID- 3046905 TI - [The official possibilities for regulation and avoidance of residues and contamination in pork]. PMID- 3046906 TI - [Endogenous opioid peptides: source, occurrence and significance in reproduction]. PMID- 3046907 TI - Developmental patterning of carbohydrate antigens during early embryogenesis of the chick: expression of antigens of the poly-N-acetyllactosamine series. AB - This report describes a striking temporal and spatial patterning of specific carbohydrate sequences in the developing chick embryo. By using oligosaccharide sequence-specific monoclonal antibodies as immunohistochemical reagents in conjunction with neuraminidase, it was possible to visualize the occurrence, as well as the changes in distribution, of oligosaccharides of the poly-N acetyllactosamine series. These were (a) long-chain unbranched sequences reactive with anti-i Den, (b) long-chain branched sequences reactive with anti-I Step and (c) short-chain branched sequences reactive with anti-I Ma and (d) their sialylated forms. The salient observations with serial sections of embryos from the unincubated to the 17th stage were as follows. (1) A pronounced anteroposterior patterning appeared during neuroectodermal development, such that the long-chain unbranched and long-chain branched sequences, which were abundant on the ectoderm of the earlier stages, were replaced by short-chain branched sialo-oligosaccharides in the developing brain and anterior neural tube. (2) A striking anteroposterior and mediolateral patterning developed in the subectodermal extracellular spaces. The long-chain linear and short-chain non sialylated sequences demarcated regions favourable for migration of the lateral plate mesoderm. (3) A distinction was made between the dorsal and ventral routes of the trunk neural crest in that the extracellular matrix of the dorsal route only was associated with long-chain linear and short-chain sialylated branched sequences. (4) A circumscribed perinotochordal distribution of the short-chain sialylated branched sequences was observed in the region of the future centra of the vertebrae. (5) An abundance of long-chain linear and long-chain sialylated branched structures was detected in primordial germ cells which permitted their identification during migration. These observations suggest that oligosaccharides of the poly-N-acetyllactosamine series may have roles as short-range, region specific information factors during morphogenetic events that take place in the developing embryo, and they open the way to the search for recognition proteins (e.g. endogenous lectins) specific for each of these oligosaccharide structures. PMID- 3046908 TI - 'Early' mammalian myoblasts are resistant to phorbol ester-induced block of differentiation. AB - Mesenchymal cells were isolated from somites and limbs of mouse embryos at different developmental stages. When grown in tissue culture, some of the cells underwent muscle differentiation as indicated by synthesis of sarcomeric myosin, acetylcholine receptor and, in the case of limb cells, fusion into multinucleated myotubes. When the tumour promoter 12-O-tetradecanoyl phorbol 13-acetate (TPA) was added to these cultures, it caused differential effects, depending upon the age of the embryo from which cells were isolated. In cultures of somites or limb bud from embryos up to 12 days post coitum, TPA did not interfere with the appearance of differentiated muscle cells. When TPA was added to cultures from older embryos, it inhibited muscle differentiation with an efficiency which increased with the age of the embryo, reaching about 90% inhibition at 15 days. After this period, a new population of myogenic cells appeared in the limb, which were able to differentiate in the presence of TPA and represented the great majority of myoblasts after day 18 of embryonic development. The simplest interpretation of these data can be based on the existence of three major classes of myogenic cell precursors, which appear sequentially during muscle histogenesis: 'early' myoblasts, which appear resistant to tumour promoters; 'late' myoblasts, whose differentiation is inhibited by tumour promoters and 'satellite' cells which, like early myoblasts, show no sensitivity to TPA. PMID- 3046909 TI - Expression and segregation of nucleoplasmin during development in Xenopus. AB - The spatial segregation of informational molecules in the unfertilized egg and embryo has been hypothesized to be a necessary phenomenon for the normal progression of development leading to the determination of cellular phenotypes. This study describes the selection of a monoclonal antibody (Mab: 2G6) that identifies an antigen (Ag: 2G6) which is localized in the germinal vesicle of oocytes and has a discrete pattern of inheritance during embryogenesis. The antigen displayed biochemical and physical characteristics very similar to nucleoplasmin, which is the histone-binding and nucleosome-assembly protein previously described. Immunoblot analysis with purified oocyte nucleoplasmin confirmed this relationship. Indirect immunofluorescence was used to study the temporal expression and spatial distribution of nucleoplasmin. From early cleavage stages through gastrulation, it is preferentially localized in nuclei of blastomeres at the animal pole. By tadpole stages, it was detected only in nuclei of postmitotic cells of the central nervous system and in nuclei of striated muscle. It was not detected in adult tissues. Western blot analysis during embryogenesis revealed at least five immunologically related polypeptides that displayed distinct patterns of expression during development. The different species observed most likely represent different levels of phosphorylation of nucleoplasmin. The more acidic forms, known to be more active in nucleosome assembly, were present during cleavage stages. Analysis of labelled oocyte proteins by two-dimensional immunoblots and autoradiography revealed that synthesis of nucleoplasmin was first detected in stage-2 oocytes, reached 60% maximum levels at stage 3, peaked at stage 4 and was undetectable in stage-6 oocytes. The amount of nucleoplasmin message present does not follow a similar pattern during oogenesis. These results suggest that the message undergoes pronounced changes in translational efficiency during oogenesis. A comparative immunoblot analysis using proteins from a variety of adult tissues revealed that, whereas the polyclonal antisera against amphibian vitellogenic oocyte nucleoplasmin recognized several different, tissue-specific polypeptides, two different monoclonal antibodies (Mab: b7-1D1, Mab: 2G6) failed to recognize any of the adult tissues tested. We conclude that nucleoplasmin is a family of closely related proteins with distinct embryonic and adult members. PMID- 3046910 TI - Genetics of sex determination in man and mouse. AB - The cytological evidence has revealed a visible mechanical basis for the production of males and females in equal numbers and irrespective of external conditions (Wilson, 1909). PMID- 3046911 TI - Expression of nuclear lamins during mouse preimplantation development. AB - The expression of nuclear lamins during mouse preimplantation development was studied by immunofluorescence, immunoblotting and immunoprecipitation. Two sera were used, specific either for lamin B or lamins A and C. Both sera gave a positive staining of the nuclear periphery throughout preimplantation development (fertilized eggs to late blastocysts). Immunoblots revealed that the three lamins were present in eggs and blastocysts. However, lamin A from eggs was found to have a higher apparent Mr than lamin A from blastocysts and other mouse cells. Using immunoprecipitation, synthesis of lamin A was detected in eggs while synthesis of lamin B was detected in 8-cell embryos and blastocysts, indicating that at least some of the lamins used during early development do not come from a store in the egg. These results are discussed in relation to the possible role of lamins during cell differentiation. PMID- 3046912 TI - [Surgical management and sequelae in breast cancer patients]. PMID- 3046913 TI - [Oral treatment of infantile diarrhea]. PMID- 3046914 TI - [Thromboembolic complications during pregnancy]. PMID- 3046915 TI - [Relapsing polychondritis]. PMID- 3046917 TI - The laryngeal mucosa in voice production. PMID- 3046916 TI - [Diagnosis of ulcer etiology]. PMID- 3046918 TI - Visual methods for the office diagnosis of voice disorders. PMID- 3046919 TI - Clinical aerodynamics for the evaluation and management of voice disorders. PMID- 3046920 TI - Trimethoprim (monotrim) compared with trimethoprim-sulphonamide in the treatment of bacterial urinary tract infection. PMID- 3046921 TI - Epignathus: a report of two cases and a review of literature. PMID- 3046922 TI - A prospective study of glucose profiles, insulin antibody levels and beta cells secretory patterns in non-obese Ugandan diabetic subjects. PMID- 3046923 TI - Evidence for two cellular pathways of renin secretion by the mouse submandibular gland. AB - The pathway of renin secretion has been defined in the mouse submandibular gland (SMG). Renin is first synthesized as a prorenin, rapidly cleaved to a one-chain renin, and then very slowly processed to a two-chain form which is stored in mature granules. In pulse-labeling experiments of minced SMG, the swift appearance in the culture medium of radiolabeled one-chain renin, before granule formation, suggested that this form was secreted by a constitutive pathway independent of the granules, possibly directly from the Golgi. Supporting this hypothesis, phenylephrine, which stimulates the secretion of granules, causes a 4 fold increase in the two-chain renin, with little or no effect on the secretion of one-chain renin. Confirming evidence for the existence of the constitutive pathway was provided by the action of monensin, an ionophore that inhibits transport through the Golgi. Monensin inhibited the appearance of radiolabeled newly synthesized renin in the granules and medium while causing accumulation of the newly synthesized one-chain renin in the microsomes. Analysis of secreted renin by Western blot showed that monensin selectively inhibited the secretion of one-chain renin while not affecting the secretion of the stored two-chain renin. Taken together, our data suggest that one-chain renin is primarily secreted soon after synthesis by a pathway that bypasses the granules, while two-chain renin is secreted predominantly from the granules. PMID- 3046924 TI - Inhibitory effects of a gonadotropin-releasing hormone agonist on the luteal synthesis of progesterone, estradiol receptors, and prolactin surges during early pregnancy. AB - Our recent studies demonstrated that the continuous administration of a GnRH agonist (GnRH-Ag; WY 40972) induces abortion in rats by suppressing plasma progesterone (P) levels within 24 h. This fall in P levels is not accompanied by a fall in ovarian venous plasma testosterone (T) or estradiol (E) levels. In this study an attempt was made 1) to determine whether the suppression of P by GnRH-Ag is due to decreased estrogen present in the corpora lutea (CL) and/or a decrease in luteal receptors of E, and 2) to investigate the effects of GnRH-Ag on the nocturnal surges of PRL. Rats were treated continuously on days 7-11 of pregnancy with 5 micrograms/day GnRH-Ag using an osmotic minipump. Ovarian blood samples were obtained on day 8; at autopsy CL were harvested and incubated with medium 199 for 4 h at 37 C under an atmosphere of 95% oxygen-5% carbon-dioxide. Additional rats were killed on day 8 or 10; CL were isolated from the ovary and pooled within the group for the measurement of nuclear and cytosolic E receptors. In other experiments, on days 8, 9, 10, and 11 of pregnancy, blood samples (0.3 ml) were collected via an indwelling intraatrial Silastic cannula at 0330, 0500, or 0600 h for the measurement of PRL and P. While the net synthesis of P by CL in the GnRH-Ag-treated rats decreased to 48 +/- 12 from 224 +/- 47 ng/CL in controls, T and E levels were not different from their respective control values. Steroid levels in ovarian venous plasma reflected a similar response. Nuclear E receptors levels were 82 and 80 in controls and 39 and 41 fmol/mg DNA in the treated group on days 8 and 10, respectively. Nocturnal surges of PRL in plasma were detected at 0330 h in controls as well as in treated rats. However, plasma PRL levels at 0330 h were 101 +/- 24, 120 +/- 22, 196 +/- 40, and 103 +/- 13 ng/ml in controls and 44 +/- 8, 50 +/- 10, 29 +/- 13, and 20 +/- 9 in the GnRH-Ag treated group on days 8, 9, 10, and 11, respectively. These results suggest that GnRH-Ag has no effect on the ability of the luteal synthesis of T and E and that the antipregnancy effect of GnRH-Ag may be at the level of the CL due to the direct inhibitory effect of GnRH-Ag on the luteal synthesis of P, which, in turn, results in decreased nocturnal surges of PRL and a fall in E receptors in the CL. Alternatively, GnRH-Ag treatment could suppress ovarian or luteal receptors for PRL, which, in turn, lower luteal E receptors, leading to a fall in luteal synthesis and release of P. PMID- 3046925 TI - Gonadotropin-releasing hormone induces oscillatory membrane currents in rat gonadotropes. AB - Electrophysiological studies were performed to characterize membrane currents of rat gonadotropes under basal conditions and after exposure to secretagogues. Gonadotropes were identified in primary cultures of rat anterior pituitaries by a reverse hemolytic plaque assay. Giga-seal patch clamp recording with the cell attached configuration was used to monitor membrane currents in these cells. Spontaneous spikes in basal current were seen. These were blocked by methoxyverapamil and probably reflect Ca2+-dependent action potentials. Brief GnRH stimulation induced slow oscillatory changes in membrane current that evolved into a series of large amplitude inward pulses after about 8 min. Treatment with TRH had no effect, and depolarization with K+ led to delayed inward currents without any oscillatory behavior. Under conditions of Ca2+ channel blockade, GnRH stimulation did not induce pulses of inward current, but did lead to oscillatory activation of a small conductance ion channel apparently selective for K+. Taken together these results suggest that GnRH induces oscillations in intracellular Ca2+ and that these oscillations are controlled by biochemical processes. PMID- 3046926 TI - Interaction of sympathomimetics and insulin with hepatic glucose production by isolated perfused rat livers: effects of continuous versus pulsatile infusion. AB - To elucidate the efficacy of continuous vs. intermittent exposure to epinephrine, phenylephrine, and insulin, hepatic glucose production was monitored in isolated perfused rat livers (means +/- SE, n = 6 each). To this end livers of fed rats were perfused with 5 mM glucose Krebs-Ringer buffer in a nonrecirculating system. Using this model it was shown that intermittent exposure (3 min on/off period, dose reduction -50%) to epinephrine (0.4 microM, alpha + beta-agonist) and phenylephrine (5 microM, alpha-agonist) elicited an almost identical rise in hepatic glucose production [epinephrine: 0.72 +/- 0.08 mmol/(86 min X 100 g BW); phenylephrine: 0.68 +/- 0.07 mmol/(86 min X 100 g BW) as their continuous administration (epinephrine: 0.78 +/- 0.06 mmol/(86 min X 100 g BW); phenylephrine: 0.74 +/- 0.09 mmol/(86 min X 100 g BW)]. Inhibition by insulin (100 mU/liter) given either continuously or intermittently (3 min on/off intervals; dose reduction -50%) was equipotent for epinephrine- and phenylephrine stimulated hepatic glucose production. When the off period was doubled to 6 min, thereby reducing the total insulin dose to 33%, no significant suppression of epinephrine- and phenylephrine-stimulated hepatic glucose production was observed. From this we conclude that 1) the effect on hepatic glucose production of pulsatile (3 min on/off, dose reduction 50%) and continuous administration is equipotent for the respective action of epinephrine, phenylephrine as well as of insulin; and 2) insulin is more effective (P less than 0.02) in inhibiting hepatic glucose production stimulated by an alpha-agonist (phenylephrine; 5.0 microM) than in counteracting alpha + beta-agonist action (epinephrine; 0.4 microM). The characteristics of hepatic glucose release as stimulated by alpha- and/or beta-adrenergic agonists and its inhibition by continuously or intermittently infused insulin were simulated and described by a computer model. Thereby, no qualitative difference could be demonstrated in alpha- vs. beta adrenergic agonists action on stimulated hepatic glucose production. PMID- 3046927 TI - Inhibition of parathyroid hormone bioactivity by human parathyroid hormone (PTH) (3-84) and PTH-(8-84) synthesized in Escherichia coli. AB - Human PTH (hPTH)-(3-84) and hPTH-(8-84) were synthesized in Escherichia (E.) coli when the cells were transformed with a multicopy plasmid in which the transcription of human preproPTH cDNA is directed by the E. coli lac promoter. PTH fragments were extracted from cells and purified by reverse phase HPLC. PTH bioactivity and PTH antagonist activity were estimated in a renal cytochemical bioassay. hPTH-(3-84) and hPTH-(8-84) exhibited less than 1% and less than 0.1%, respectively, of the biological activity of synthetic hPTH-(1-84). hPTH-(8-84) had 1% of the PTH inhibitory activity of synthetic [Nle8,18,Tyr34]bovine PTH-(3 34)amide, whereas hPTH-(3-84) was 100 times more active as a PTH inhibitor than the synthetic bovine PTH-(3-34) analog. The latter has so far been recognized as the most potent PTH antagonist in vitro. A 5-fold molar excess of hPTH-(3-84) over hPTH-(1-84) completely blocked the biological action of intact hPTH-(1-84) in the renal cytochemical bioassay. These findings suggest that the carboxyl terminal portion of the intact hPTH-(1-84) molecule contributes importantly to inhibitor potency. PMID- 3046928 TI - Regulation of luteinizing hormone subunit biosynthesis in cultured male anterior pituitary cells: effects of gonadotropin-releasing hormone and testosterone. AB - The purpose of this study was to evaluate the direct effects of testosterone (T) on LH subunit apoprotein synthesis, glycosylation, and release by the male pituitary. Cells from 1-week castrate rats were cultured for 48 h in steroid-free medium, followed by 48 h in medium with or without 10 nM T. The cells were then incubated for 2, 4, 6, 8, or 12 h in medium containing [35S]methionine (35S-Met) or [3H]glucosamine (3H-Gln), with or without 1 nM GnRH (Exp 1) or in medium containing precursors with or without 10 nM T and/or 1 nM GnRH (Exp 2). Radiolabeled precursor incorporation into LH subunits was determined by immunoprecipitation, followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In Exp 1, precursor incorporation into total protein (TP) and LH subunits increased linearly over time for at least 8 h. GnRH did not affect precursor incorporation into total protein or 35S-Met labeling of LH subunits, but stimulated a linear time-dependent accumulation of 3H-Gln into total (cells plus media) LH subunits and release of radioimmunoassayable LH into the medium. Based on these results, the effects of T on LH subunit biosynthesis (with or without GnRH) were studied during an 8-h incubation. In Exp 2, GnRH enhanced total 3H-Gln (but not 35S-Met) incorporation into both LH subunits. GnRH stimulated the release of 35S-Met LH alpha and 3H-Gln LH subunits and increased the relative glycosylation of secreted LH subunits without altering the relative glycosylation of intracellular LH subunits. T inhibited radioimmunoassayable LH release and incorporation of both precursors into total and secreted LH subunits (with or without GnRH). However, only the relative glycosylation of secreted LH alpha was reduced by T (with or without GnRH). These data indicate that T acts directly at the pituitary to inhibit LH subunit apoprotein synthesis and selectively inhibit LH alpha glycosylation. Further, these data support the hypothesis that changes in LH glycosylation may be one of the ways by which GnRH and T regulate LH release. PMID- 3046929 TI - Mobilization of Ca2+ from intracellular stores of pancreatic beta-cells by the calcium store blocker TMB-8. AB - TMB-8 has been used experimentally in many cell types, including endocrine cells, because of its ability to block the efflux of Ca2+ from intracellular stores without affecting influx. Unexpectedly, TMB-8 potentiates stimulated insulin release from pancreatic islets, a process believed to be dependent on the level of cytosolic Ca2+. In the present study, while having no effect on basal insulin release (in the presence of 2.8 mM glucose), TMB-8 (10, 30, and 100 microM) caused a concentration-dependent increase in 45Ca2+ efflux from 45Ca2+-preloaded islets. TMB-8 (100 microM) stimulated 45Ca2+ efflux even in the absence of extracellular Ca2+. In the presence of 5.6 mM glucose, TMB-8 (30 and 100 microM) potentiated insulin release and again increased 45Ca2+ efflux in a concentration dependent manner. Similarly, insulin release stimulated by isobutylmethylxanthine (IBMX) was potentiated significantly, and IBMX-stimulated 45Ca2+ efflux was increased by the simultaneous introduction of 30 microM TMB-8. Thus, in pancreatic islets, TMB-8 appears to mobilize Ca2+ from intracellular stores, rather than inhibit the efflux as has been commonly accepted. In further studies, using insulin-secreting beta-cells of the RINm5F cell line, TMB-8 was shown to increase the cytosolic Ca2+ concentration in the presence and absence of extracellular Ca2+. This confirmed that mobilization of intracellular Ca2+ was occurring in the pancreatic beta-cell in response to TMB-8. Furthermore, a rise in cytosolic Ca2+ of not more than 10 nM (as induced with KCl) was found to mimick the effect of TMB-8 in conjunction with IBMX. No additional effect of TMB 8 to alter Ca2+ handling at the plasma membrane was found when 45Ca2+ uptake experiments were performed. Therefore, the paradoxical mobilization of beta-cell Ca2+ by TMB-8 appears to be a sufficient explanation for its potentiating effect on the rate of insulin secretion. PMID- 3046930 TI - Long term improvement of glucose homeostasis by vanadate treatment in diabetic rats. AB - The trace element vanadium (V) exerts insulin-like effects in vitro. The present study examined its effects on glucose homeostasis in rats made diabetic by streptozotocin. Na3VO4 (0.2 or 0.5 mg/ml) was administered ad libitum in drinking water. Fed plasma glucose levels (approximately 26 mmol/liter) fell by 30% and 56% after 5 days of treatment with the low (VO.2) and high (VO.5) concentrations of vanadate, respectively. This decrease was not due to a rise in peripheral insulin levels and persisted for more than 2 months. Daily glucosuria was decreased by 60% and 85% in VO.2 and VO.5 rats, respectively. Tolerance of the rats to oral or iv glucose was also considerably improved by vanadate; integrated glucose responses were about 55% and 75% lower in VO.2 and VO.5 rats than in controls, and the differences were not due to restoration of insulin release. Compared to nondiabetic rats, pancreatic insulin reserves amounted to 1% in untreated rats, 3% in VO.2 rats, and 6% in VO.5 rats after 9 weeks of treatment. Liver, but not muscle, glycogen was increased by vanadate. Despite improvement of their diabetic state, vanadate-treated rats did not gain more weight than untreated rats. Their food intake (corrected for urinary glucose losses) was decreased by about 25%. No signs of altered kidney or liver function were observed in rats receiving vanadate. In conclusion, vanadate markedly improves glucose homeostasis in streptozotocin-diabetic rats by an insulin-like mechanism, but does not reproduce the anabolic effects of the hormone. PMID- 3046931 TI - The mouse protein synthesis initiation factor 4A gene family includes two related functional genes which are differentially expressed. AB - We have cloned and characterized a family of mouse genomic sequences hybridizing to mouse cDNA probes coding for eIF-4A, one of the protein synthesis initiation factors involved in the binding of mRNA to the ribosome. We estimate that there is a total of approximately 9-13 eIF-4A pseudogenes. We also found an eIF-4A intronless retroposon which, when compared to the cDNA, contains a single nucleotide difference. This possibly functional gene contains a mouse repetitive B1 element integrated in the promoter region. Furthermore, we have cloned two intron-containing eIF-4A genes (termed eIF-4AI and eIF-4AII). The eIF-4AII gene codes for a previously unknown form of eIF-4A. Northern blot hybridization with RNA from several mouse organs shows a variation in eIF-4AI expression within a factor of 7. In contrast, relative to liver, eIF-4AII expression is 20- to 30 times higher in brain and kidney, 10- to 17-fold higher in lung and heart, and is about equally abundant in liver, spleen and thymus. These data suggest that the relative efficiency of protein synthesis initiation for different mRNAs, as reflected by discrimination in messenger 5'-terminal cap recognition and binding to ribosomes, varies in different tissues. PMID- 3046932 TI - Sequence analysis of ARS elements in fission yeast. AB - Chromosomal DNA of Schizosaccharomyces pombe contains sequences with properties analogous to ARS elements of Saccharomyces cerevisiae. Following Sau3A fragmentation of the S. pombe genome we have recovered a number of such fragments in an M13-based shuttle vector, suitable for subsequent sequence analysis. The complete nucleotide sequence has been obtained for eight ARS+ inserts derived from the Sau3A cloning and for the ARS present in pFL20 isolated previously by Losson and Lacroute (Cell, 32, 371-377, 1983). The Sau3A clones are single fragments between 0.8 and 1.8 kb. No ARS+ clones smaller than this were recovered even though the average size Sau3A fragment in S. pombe is approximately 200-300 bp. The sequence analysis revealed that all clones are AT-rich (69-75% A + T residues), and all contain a particularly AT-rich 11 bp core element represented by the consensus sequence 5' (A/T)PuTT-TATTTA(A/T) 3'. Deletion mapping indicates that the consensus in all cases is in the vicinity of a functional ARS domain. However precise excision of the consensus by in vitro mutagenesis has little effect on ARS activity as judged by the transformation assay. We argue that the association of the consensus with the ARS domain occurs too reproducibly to be explained by chance alone. We suggest that although it may not be essential for the extrachromosomal maintenance of plasmids in S. pombe, the consensus does have a function in situ in the chromosome and thus is always present as a cryptic sequence in the isolated ARS element. PMID- 3046933 TI - Effects of histone H4 depletion on the cell cycle and transcription of Saccharomyces cerevisiae. AB - We have constructed a yeast strain (UKY403) in which the sole histone H4 gene is under control of the GAL1 promoter. This allows the activation of H4 mRNA synthesis on galactose and its repression on glucose. UKY403 cells, pre synchronized in G1 with alpha-mating factor, have been used to show that glucose treatment results in the loss of approximately half the chromosomal nucleosomes. This depletion is only partially reversible when the H4 gene is reactivated on galactose. It was found that the resultant lethality manifests itself first in S phase, the period of nucleosome assembly, but leads to highly synchronous arrest in G2 and a virtually complete block in chromosomal segregation. Histone H4 depleted chromatin was analyzed for its efficiency as a template for all three RNA polymerases. Using pulse-labeling, we find no evidence for altered transcription by RNA polymerase I (25S, 18S and 5.8S rRNAs) or RNA polymerase III (5S rRNA, tRNAs). Northern blot analysis was used to measure levels of RNA polymerase II transcripts. There was little effect on the activation or repression of the CUP1 chelatin gene. While there may be some decrease in the level of certain mRNAs (e.g. HIS4, ARG4) other message levels (HIS3, TRP1) show little change upon glucose repression. Therefore, nucleosome loss certainly does not have a general effect on transcription. PMID- 3046934 TI - Depletion of histone H4 and nucleosomes activates the PHO5 gene in Saccharomyces cerevisiae. AB - We have previously constructed a yeast strain (UKY403) whose sole histone H4 gene is under control of the GAL1 promoter. This yeast arrests in G2 upon glucose treatment as a result of histone H4 depletion. The yeast PHO5 gene contains phase nucleosomes covering promoter (UAS) sequences in the PHO5 repressed state and it has been suggested that nucleosomes prevent the binding of positively acting factors to these UAS sequences. Using UKY403 we examined the length of polynucleosomes and nucleosome phasing in the PHO5 upstream region by the use of micrococcal nuclease and indirect end-labeling. It was found that glucose arrest led to a severe disruption in PHO5 chromatin structure and that most nucleosomes had their position altered or were lost from the PHO5 promoter region. Cell undergoing nucleosome depletion synthesized large quantities of accurate PHO5 transcripts even under repressive, high inorganic phosphate conditions. Histone H4 depletion did not appear to affect the repression or activation of another inducible yeast gene, CUP1. Arrest with landmarks in early G1 (in the cell division cycle mutant cdc28) or in various stages of G2 (in cdc15, cdc17 and cdc20) does not activate PHO5; nor does arrest due to chromosome topology changes (in top2 or the top1top2 topoisomerase mutants). cdc14, which has its arrest landmark at a similar point in the cell cycle as cdc15, does derepress PHO5. However, since it also leads to derepression of CUP1 it is probably functioning through an independent mechanism. Therefore, our data suggest that nucleosomes regulate PHO5 transcription. PMID- 3046935 TI - Secretion in yeast: in vitro analysis of the sec53 mutant. AB - Of central importance to studying protein translocation via a combined genetic and biochemical approach is the in vitro analysis of yeast conditionally-lethal secretory mutants. Analysis of sec53 presented an opportunity not only to see if mutants could be examined in recently developed yeast in vitro translocation systems, but also to characterize further the nature of this mutant originally postulated to be defective in protein translocation. Membranes from sec53 were capable of translocating and glycosylating nascent prepro-alpha-factor in vitro in both sec53 and wild-type lysates at temperatures that were non-permissive for growth of the mutant cells. These results suggested that the Sec53 protein does not function directly in the translocation and glycosylation of prepro-alpha factor. To examine this point further, we isolated membranes from sec53 cells that had been grown at the non-permissive temperature prior to disruption. In such cases, regardless of assay temperature, membranes from sec53 cells efficiently translocated but failed to glycosylate prepro-alpha-factor in vitro. The in vitro phenotype of sec53 could be mimicked by isolating rough microsomes from wild-type cells that had been grown for 1 h in the presence of tunicamycin. Together, these results demonstrate that sec53 is not defective in translocation, rather in assembly of the dolichol-oligosaccharide substrate needed for N-linked glycosylation. PMID- 3046936 TI - A major 125-kd membrane glycoprotein of Saccharomyces cerevisiae is attached to the lipid bilayer through an inositol-containing phospholipid. AB - A number of plasma membrane glycoproteins of mammalian and protozoan origin are released from cells by phosphatidylinositol-specific phospholipase C. Some of these proteins have been shown to be attached to the lipid bilayer via a covalently linked, structurally complex glycophospholipid. Here we establish the existence of similarly linked glycoproteins in the yeast Saccharomyces cerevisiae. The most abundant of these is a tightly membrane-bound glycoprotein of 125 kd. The detergent-binding moiety of this protein can be removed by phosphatidylinositol-specific phospholipase C of bacterial origin or from Trypanosoma brucei. Metabolic labeling indicates that the protein contains covalently attached fatty acid and inositol. It also contains the cross-reacting determinant (CRD), an antigen found previously on the glycophospholipid anchor of protozoan and mammalian origin. Treatment of the protein with endoglycosidases F and H results in a 95-kd species. In the secretion mutant sec18, grown at 37 degrees C, the vesicular transport of glycoproteins is reversibly blocked between the rough endoplasmic reticulum and the Golgi apparatus. We find that sec18 cells, when grown at 37 degrees C, do add phospholipid anchors to newly synthesized glycoproteins. This indicates that these anchors are added in the rough endoplasmic reticulum. PMID- 3046937 TI - Roles of two DNA-binding factors in replication, segregation and transcriptional repression mediated by a yeast silencer. AB - The HMR E silencer is required for SIR-dependent transcriptional repression of the silent mating-type locus, HMR. The silencer also behaves as an origin of replication (ARS element) and allows plasmids to replicate autonomously in yeast. The replication and segregation properties of these plasmids are also dependent on the four SIR genes. We have previously characterized two DNA-binding factors in yeast extracts that recognize specific sequences at the HMR E silencer. These proteins, called ABFI (ARS-Binding Factor) and GRFI (General Regulatory Factor), are not encoded by any of the SIR genes. To investigate the biological roles of these factors, single-base-pair mutations were constructed in both binding sites at the HMR E silencer that were no longer recognized by the corresponding proteins in vitro. Our results indicate that the GRFI-binding site is required for the efficient segregation of plasmids replicated by the HMR E silencer. SIR dependent transcriptional repression requires either an intact ABFI-binding site or GRFI-binding site, although the GRFI-binding site appears to be more important. A double-mutant silencer that binds neither ABFI nor GRFI does not mediate transcriptional repression of HMR. The replacement of HMR E with a chromosomal origin of replication (ARS1) allows partial SIR-dependent transcriptional repression of HMR, indicating a role for replication in silencer function. Together, these results suggest that the SIR proteins influence the properties of the HMR E silencer through interactions with other DNA-binding proteins. PMID- 3046940 TI - Sequence of the Saccharomyces cerevisiae CTA1 gene and amino acid sequence of catalase A derived from it. AB - The nucleotide sequence of a 2785-base-pair stretch of DNA containing the Saccharomyces cerevisiae catalase A (CTA1) gene has been determined. This gene contains an uninterrupted open reading frame encoding a protein of 515 amino acids (relative molecular mass 58,490). Catalase A, the peroxisomal catalase of S. cerevisiae was compared to the peroxisomal catalases from bovine liver and from Candida tropicalis and to the non-peroxisomal, presumably cytoplasmic, catalase T of S. cerevisiae. Whereas the peroxisomal catalases are almost colinear, three major insertions have to be introduced in the catalase T sequence to obtain an optimal fit with the other proteins. Catalase A is most closely related to the C. tropicalis enzyme. It is also more similar to the bovine liver catalase than to the second S. cerevisiae catalase. The differences between the two S. cerevisiae enzymes are most striking within four blocks of amino acids consisting of a total of 37 residues with high homology between the three peroxisomal, but low conservation between the S. cerevisiae catalases. The results obtained indicate that the peroxisomal catalases compared have very similar three-dimensional structures and might have similar targeting signals. PMID- 3046938 TI - Alternative pathways for the in vivo repair of O6-alkylguanine and O4 alkylthymine in Escherichia coli: the adaptive response and nucleotide excision repair. AB - The in vivo removal of three different O-alkylated bases from DNA was measured in Escherichia coli. Using monoclonal antibodies specific for O6-methylguanine, O6 ethylguanine and O4-ethylthymine we have monitored the removal of these lesions from six different strains to assess the relative contributions of the adaptive response and of nucleotide excision repair. During the first hour after DNA alkylation, O6-methylguanine, O6-ethylguanine and O4-ethylthymine lesions were repaired almost exclusively by nucleotide excision, except when the adaptive response was being constitutively expressed. In wild-type E. coli the adaptive response began to contribute to O6-methylguanine repair about one hour after alkylation, the time required for the full induction of the ada DNA methyltransferase. In contrast, the adaptive response did not play such a large role in the repair of O6-ethylguanine and O4-ethylthymine in wild-type E. coli, presumably because DNA ethylation damage is a poor inducer of the adaptive response; possible reasons for this poor induction are discussed. The repair of all three O-alkylated lesions was virtually absent in ada- uvr- bacteria suggesting that no alternative pathway is available for their repair, at least during the first two hours after alkylation. When the repair of O-alkylated bases was compromised by an ada- or by a uvr- mutation, the bacteria became more sensitive to alkylation induced killing and mutation. PMID- 3046939 TI - In vivo and in vitro identity of site specific cleavages in the 5' non-coding region of ompA and bla mRNA in Escherichia coli. AB - The bla and ompA gene transcripts were used as substrates to probe Escherichia coli extracts for ribonucleolytic activities. A site specific endoribonucleolytic activity was identified that cleaves ompA and bla mRNA. The cleavages occur in vitro and in vivo. For both the bla and ompA mRNA most of the cleavage sites which were identified map in the 5' non-coding region. The cleavages of the ompA transcript have been previously suggested to regulate the growth rate dependent stability of this mRNA. Thus we propose that the identified endoribonucleolytic activity may be involved in the degradation of mRNA. Analysis of mutants revealed that the cleavages are mediated by endonucleases which do not seem to be identical to RNase III, RNase E or RNase P. PMID- 3046941 TI - Comparison of the amino acid sequences of the transacylase components of branched chain oxoacid dehydrogenase of Pseudomonas putida, and the pyruvate and 2 oxoglutarate dehydrogenases of Escherichia coli. AB - The nucleotide sequence of bkdB, the structural gene for E2b, the transacylase component of branched-chain-oxoacid dehydrogenase of Pseudomonas putida has been determined and translated into its amino acid sequence. The start of bkdB was identified from the N-terminal sequence of E2b isolated from branched-chain oxoacid dehydrogenase of the closely related species, P. aeruginosa. The reading frame was composed of 65.5% G + C with 82.3% of the codons ending in G or C. There was no intergenic space between bkdA2 and bkdB. No codons requiring minor tRNAs were utilized and the codon bias index indicated a preferential codon usage. The bkdB gene encoded 423 amino acids although the N-terminal methionine was absent from E2b prepared from P. aeruginosa. The relative molecular mass of the encoded protein was 45,134 (45,003 minus methionine) vs 47,000 obtained by SDS/polyacrylamide gel electrophoresis. There was a single lipoyl domain in E2b compared to three lipoyl domains in E2p, and one domain in E2o, the transacylases of pyruvate and 2-oxoglutarate dehydrogenases of Escherichia coli respectively. There was significant similarity between the lipoyl domain of E2b and of E2p and E2o as well as between the E1-E2 binding domains of E2b, E2p and E2o. There was no similarity between the E3 binding domain of E2b to E2p and E2o which may reflect the uniqueness of the E3 component of branched-chain-oxoacid dehydrogenase of P. putida. The conclusions drawn from these comparisons are that the transacylases of prokaryotic pyruvate, 2-oxoglutarate and branched-chain oxoacid dehydrogenases descended from a common ancestral protein probably at about the same time. PMID- 3046942 TI - Purification and characterization of rat-liver cytosolic epoxide hydrolase. AB - Rat liver cytosolic epoxide hydrolase has been purified and characterized. The enzyme was purified from tiadenol-induced rat liver 540-fold with respect to trans-stilbene oxide as a substrate. Similar purification was obtained with the substrates trans-beta-ethyl styrene oxide and styrene 7,8-oxide, the specific activities decreasing in the order trans-beta-ethyl styrene oxide greater than styrene 7,8-oxide greater than trans-stilbene oxide. The enzyme exerts highest activity at pH 7.4 Km and Vmax of the pure enzyme for trans-stilbene oxide were 1.7 microM and 205 nmol x min-1 x mg protein-1 respectively. With trans-stilbene oxide as a substrate, the inhibition by organic solvents (2.5% by vol.) increased in the order ethanol less than methanol less than acetone less than isopropanol = N,N-dimethyl formamide less than acetonitrile less than tetrahydrofuran. The native enzyme, with a molecular mass of 120 kDa, consists of two 61-kDa subunits. Digestion of rat liver cytosolic and microsomal epoxide hydrolase by three proteases resulted in markedly different peptide maps. Western-blot analysis with antiserum against rat liver cytosolic epoxide hydrolase revealed a single band with the purified enzyme, and with liver cytosol from control and clofibrate induced rats. No cross-reactivity was observed with purified rat microsomal epoxide hydrolase or microsomes. A positive reaction at the same molecular mass was obtained with liver cytosol of mouse, guinea pig, Syrian hamster and New Zealand white rabbit but not with that of green monkey. PMID- 3046943 TI - Chorismate mutase:prephenate dehydratase from Acinetobacter calcoaceticus. Purification, properties and immunological cross-reactivity. AB - The bifunctional P protein (chorismate mutase: prephenate dehydratase) from Acinetobacter calcoaceticus has been purified. It was homogeneous in polyacrylamide gels and was more than 95% pure on the basis of the immunostaining of purified P protein with the antibodies raised against the P protein. The native enzyme is a homodimer (Mr = 91,000) composed of 45-kDa subunits. A twofold increase in the native molecular mass of the P protein occurred in the presence of L-phenylalanine (inhibitor of both activities) or L-tyrosine (activator of the dehydratase activity) during gel filtration. Chorismate mutase activity followed Michaelis-Menten kinetics with a Km of 0.55 mM for chorismate. L-Phenylalanine was a relatively poor non-competitive inhibitor of the mutase activity. The chorismate mutase activity was also competitively inhibited by prephenate (reaction product). Substrate-saturation curves for the dehydratase activity were sigmoidal showing positive cooperativity among the prephenate-binding sites. L Tyrosine activated prephenate dehydratase strongly but did not abolish positive cooperativity with respect to prephenate. L-Phenylalanine inhibited the dehydratase activity, and the substrate-saturation curves became increasingly sigmoidal as phenylalanine concentrations were increased with happ values changing from 2.0 (no phenylalanine) to 4.0 (0.08 mM L-phenylalanine). A sigmoidal inhibition curve of the dehydratase activity by L-phenylalanine gave Hill plots having a slope of -2.9. Higher ionic strength increased the dehydratase activity by reducing the positive cooperative binding of prephenate, and the sigmoidal substrate-saturation curves were changed to near-hyperbolic form. The happ values decreased with increase in ionic strength. Antibodies raised against the purified P protein showed cross-reactivity with the P proteins from near phylogenetic relatives of A. calcoaceticus. At a greater phylogenetic distance, cross-reaction was superior with P protein from Neisseria gonorrhoeae than with that from the more closely related Escherichia coli. PMID- 3046944 TI - The role of aspartic acid-49 in the active site of phospholipase A2. A site specific mutagenesis study of porcine pancreatic phospholipase A2 and the rationale of the enzymatic activity of [lysine49]phospholipase A2 from Agkistrodon piscivorus piscivorus' venom. AB - In order to probe the role of Asp-49 in the active site of porcine pancreatic phospholipase A2 two mutant proteins were constructed containing either Glu or Lys at position 49. Their enzymatic activities and their affinities for substrate and for Ca2+ ions were examined in comparison with the native enzyme. Enzymatic characterization indicated that the presence of Asp-49 is essential for effective hydrolysis of phospholipids. Conversion of Asp-49 to either Glu or Lys strongly reduces the binding of Ca2+ ions in particular for the lysine mutant but the affinity for substrate analogues is hardly affected. Extensive purification of [Lys49]phospholipase A2 from the venom of Agkistrodon piscivorus piscivorus yielded a protein which was 4000 times less active than the basic [Asp49]phospholipase A2 from this venom. Inhibition studies with p-bromophenacyl bromide showed that this residual activity was due to a small amount of contaminating enzyme and that the Lys-49 homologue itself is inactive. The results obtained both with the porcine pancreatic phospholipase A2 mutants and with the native venom enzymes show that Asp-49 is essential for the catalytic action of phospholipase A2. PMID- 3046946 TI - Assembly of amber fragments of the beta subunit of Escherichia coli RNA polymerase. AB - Immunoblotting of size-separated whole cell proteins permitted the study of protein-protein interaction. Briefly, proteins obtained from cleared cell lysates of Escherichia coli were separated by glycerol gradient centrifugation and analysed by blotting against a set of specific antibodies. We have applied this procedure to the assembly of 11 N-terminal amber fragments of the beta subunit of E. coli RNA polymerase ranging in size between 97% and 23% the length of the intact beta polypeptide (1342 amino acids). In this way, we have been able to define regions on the beta polypeptide involved in the assembly of RNA polymerase. PMID- 3046947 TI - Recurrent vascular myelopathy. Report of a case with autopsy. AB - A case of ischemic myelopathy which was marked by two clinical episodes, separated by a 4-month interval and affecting the same level of the spinal cord, is reported. A wide-ranging search through the literature shows that the recurrent type of ischemic vascular myelopathy is very rare. PMID- 3046945 TI - ATP-induced activation of the aminoacylation of tRNA by the isoleucyl-tRNA synthetase from Escherichia coli. AB - The rate of aminoacylation of tRNA catalyzed by the isoleucyl-tRNA synthetase form Escherichia coli has been measured. A steady-state kinetic analysis of the rate as a function of the concentration of ATP gave nonlinear Hanes plots. ATP behaves as an activator of the reaction. The activation is observed at a low magnesium ion concentration and in the presence of spermidine. The presence of inorganic pyrophosphate or AMP enhances the activation. The results are consistent with a mechanism in which the binding of a second molecule of ATP increases the rate of dissociation of Ile-tRNA from the enzyme. PMID- 3046948 TI - Endocrine therapy of breast cancer. AB - The aim of this literature study is to evaluate the various endocrine drugs used in the treatment of breast cancer in order to compare their therapeutic efficacy. Hormone-dependent metastasized breast cancer of post-menopausal women can be treated with four equivalent drugs: tamoxifen, megestrol, medroxyprogesterone, and--combined with steroid suppletion therapy--aminoglutethimide. Each of these drugs induces (partial) remissions in 50-60% of estrogen receptor-positive tumors. Because of its relatively few and less serious side-effects tamoxifen is the agent of first choice for soft tissue and lung metastases. Due to its potentially enhanced objective and more pronounced subjective side-effects, aminoglutethimide may be preferred in bone metastases. In view of the only minor activity of hormonal agents in liver metastases, combination chemotherapy with concomitant, sequential or alternating use of hormonal agents should be considered in this condition. Until now, only tamoxifen seems suitable for the treatment of premenopausal metastasized hormone-dependent breast cancer. In the adjuvant setting, preoperatively initiated and as soon as possible postoperatively continued endocrine therapy seems to be of the utmost importance to counteract (occult) metastases while reducing the need for extensive surgery. PMID- 3046949 TI - The role of lymphocyte function-associated antigen 1 (LFA-1) in the adherence of T lymphocytes to B lymphocytes. AB - The functional role of the LFA-1 molecule in the interaction between helper T lymphocytes and B lymphocytes was investigated using lymphocytes from patients with leukocyte adhesion deficiency, an inherited immunodeficiency characterized by a defective leukocyte expression of the LFA-1, Mac-1 (CR3) and p150,95 molecules. The ability of LFA-1- T lymphocytes to provide antigen-specific help for HLA-identical LFA-1+ B lymphocytes was reduced while their antigen-specific activation was normal. Antigen-independent conjugate formation between resting, nonactivated LFA-1- T lymphocytes and LFA-1+ B lymphocytes was impaired while LFA 1- B lymphocytes bound LFA-1+ T lymphocytes normally. Conjugate formation of activated LFA-1- T lymphocytes was mostly mediated by the CD2-LFA-3 adhesion pathway while the ICAM-1 molecule, a ligand of LFA-1, had no function. These results demonstrate that LFA-1 plays a major role in the cognate interaction between helper T lymphocytes and B lymphocytes that cannot be mediated instead by CD2 or other molecules on resting T lymphocytes. PMID- 3046950 TI - Molecular mechanisms underlying lymphocyte recirculation. I. Functional, phenotypical and morphological characterization of high endothelial cells cultured in vitro. AB - Large-scale extravasation of lymphocytes takes place in vivo under physiological conditions in lymph nodes at very specialized vascular segments called high endothelial venules (HEV). When circulating lymphocytes leave the blood, they first bind to endothelial cells of HEV (HE cells) and subsequently enter lymph nodes by crossing the endothelial lining of HEV. Although the lymphocyte-HEV interaction has recently been the subject of intense research by many laboratories, it has been studied almost exclusively by the use of the lymphocyte binding assay in which lymphocyte binding is examined on nonviable HEV present on frozen sections and, hence, no dynamic interaction between HE cells and lymphocytes has been studied. We report herein that endothelial cells of rat HEV can be grown in vitro and that the lymphocyte-HEV interaction can be studied dynamically using viable cells in culture vessels. The identification of the cultured line, termed Ax, as HE cells was based on their phenotypic, morphological, cytochemical and biochemical characteristics, and most importantly on its in vitro behavior, particularly in terms of its specific ability to interact with mature lymphocytes. Phenotypic analysis demonstrated that not only did monoclonal antibodies, known to react with HE cells, recognize the Ax but also a monoclonal antibody raised against the Ax specifically recognized HE cells in vivo, as determined by an immunoperoxidase staining of frozen sections, supporting the notion that the cell strain, Ax, is derived from HEV. This Ax, even after long-term culture (greater than 50 passages), allowed mature, but not immature, lymphocytes to bind to the cell surface and subsequently transport bound cells underneath their cytoplasm. This phenomenon was inhibited in a dose dependent manner by various reagents known to inhibit lymphocyte recirculation in vivo. The cultured line derived from HE cells should provide a means to investigate the biochemical nature of lymphocyte-HEV interaction, and to understand the molecular mechanisms underlying the large-scale lymphocyte traffic taking place in vivo. PMID- 3046951 TI - Distribution of membrane cofactor protein of complement on human peripheral blood cells. An altered form is found on granulocytes. AB - Membrane cofactor protein (MCP) of human complement is an iC3/C3b-binding glycoprotein with a characteristic two-band (63 kDa and 55 kDa) pattern on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Using affinity chromatography, it has been found on human mononuclear cells and platelets. MCP has been purified and shown to be a cofactor for the I-mediated cleavage of C3b. A rabbit polyclonal antibody was produced to the purified protein and this reagent employed to analyze the distribution of MCP on human peripheral blood cells. Flow cytometric analysis indicated that MCP is unimodally present on all platelets, granulocytes, T helper lymphocytes, T suppressor/cytotoxic lymphocytes, B lymphocytes, natural killer cells and monocytes but not erythrocytes. The presence of MCP on granulocytes was unexpected. To evaluate this, MCP was isolated by immunoprecipitation and analyzed by SDS-PAGE followed by autoradiography. The Mr of granulocyte MCP was that of a single broad band in which the typical two-band pattern could not be distinguished. Alterations in the conditions of the affinity column procedure increased the efficiency of the isolation of monocyte MCP and led to the reproducible isolation of granulocyte MCP. These results indicate that MCP of granulocytes has both structural and functional differences compared to MCP of plateletes and mononuclear cells. The wide distribution of MCP among peripheral blood cells supports the concept that MCP is important in the protection of host cells from complement-mediated damage. PMID- 3046953 TI - Effect of bromelain on kaolin-induced inflammation in rats. AB - The effects of stem bromelain on the plasma kallikrein system, bradykinin levels and plasma exudation at the inflammatory site were examined in rats with a kaolin induced inflammation of an air pouch. Bromelain (5, 7.5 mg/kg) caused a dose dependent decrease of bradykinin levels at the inflammatory site and a parallel decrease of the prekallikrein levels in sera. Plasma exudation was also reduced dose dependently. Bradykinin-degrading activity in sera was elevated after treatment with bromelain, although it was unchanged in the pouch fluid. These data indicate that bromelain inhibits plasma exudation through inhibiting the generation of bradykinin at the inflammatory site via depletion of the plasma kallikrein system. PMID- 3046952 TI - Effects of the specific platelet-activating factor antagonists, BN 52021 and BN 52063, on various experimental gastrointestinal ulcerations. AB - Platelet-activating factor (PAF) has been shown recently to induce gastrointestinal damage similar to that evoked by endotoxin, suggesting that the autacoid could be implicated in other types of gastrointestinal damage. Thus, the effects of BN 52021 and BN 52063, two specific PAF antagonists, were investigated in various experimental models of gastrointestinal damage in rats. BN 52021 and BN 52063, markedly reduced both the PAF- and endotoxin-induced alterations of the mucosa, suggesting a role for the autacoid in the latter process. BN 52021 and another unrelated PAF antagonist, triazolam, partially reduced the restraint stress-induced gastric damage in young female, but not male, rats. Similar partial protection was obtained in rats with ethanol-induced gastric damage. In contrast with atropine and ranitidine, BN 52021 did not affect the gastric hypersecretion in pylorus-ligated rats nor the aspirin-induced gastric ulcerations. The present results indicate that PAF plays a major role in the gastric damage induced by endotoxin and may also partially contribute to the gastric lesions induced by ethanol and stress. The results suggest that there is a potential therapeutic use for PAF antagonists in certain types of gastrointestinal lesions in man. PMID- 3046954 TI - Both competitive and non-competitive antagonists of N-methyl-D-aspartic acid disrupt brightness discrimination in rats. AB - Rats were trained to avoid or escape electric shocks in a symmetrical Y-maze by choosing to enter the brighter of two arms. Pretreatment with phencyclidine-like compounds disrupted brightness discrimination with greatly increased spontaneous locomotor activity between trials. The competitive antagonists of NMDA, 2-amino-7 phosphonoheptanoate (AP7) or 3-(+/-)-2-carboxypiperazin-4-yl)propyl-1-phosphonic acid (CPP) also disrupted brightness discrimination when injected into the cerebral ventricles, with no increase in movements between trials. The results suggest that the competitive antagonists of NMDA may impair sensory and cognitive functions in a manner similar to that produced by the phencyclidine-like compounds. PMID- 3046955 TI - Dextromethorphan inhibits NMDA-induced convulsions. AB - Dextromethorphan, its metabolite dextrorphan, phencyclidine, ketamine, MK-801, 3 (2-carboxypiperazin-4-yl)propyl-1-phosphonic acid and DL-2-amino-7 phosphonoheptanoic acid were evaluated for potency to antagonize N-methyl-D aspartate-induced convulsions following intraperitoneal administration using male CF-1 mice. Whereas reference anticonvulsants (e.g., phenytoin) were ineffective in this model, dextromethorphan and all competitive and noncompetitive N-methyl-D aspartate antagonists blocked seizures. The results are consistent with the interpretation that dextromethorphan elicits some of its pharmacological responses via an interaction with receptors for excitatory amino acids. PMID- 3046956 TI - Actions of nitric oxide on the release of prostacyclin from bovine endothelial cells in culture. AB - Endothelial cells release the potent vasodilator prostacyclin, as well as the highly labile endothelium-derived relaxing factor (EDRF) which mediates vascular relaxation induced by some vasodilators including acetylcholine and bradykinin. EDRF has recently been characterised as nitric oxide (NO). The effects of NO on prostacyclin release, measured as 6-keto-PGF1 alpha, from endothelial cells obtained from bovine thoracic aorta, have now been investigated. Incubation of endothelial cells in culture with bradykinin (10-100 nM) stimulated the release of 6-keto-PGF1 alpha. Pre-incubation (0.5-2 min) with NO (13-130 microM) caused a significant dose-dependent inhibition of 6-keto-PGF1 alpha release, reaching a maximum of 29 +/- 4% inhibition. Pre-incubation with superoxide dismutase (30 units ml-1) which prevents the breakdown of NO, significantly augmented the degree of inhibition, as did the selective inhibitor of cyclic GMP phosphodiesterase, M & B 22948 (5 microM), reaching 51 +/- 2% inhibition. The potentiation by M & B 22948 suggests that this inhibitory effect of high concentrations of NO is brought about by elevation of intracellular cyclic GMP levels following activation of guanylate cyclase. Whether endogenous NO is produced by endothelial cells under physiological conditions in sufficient quantities to modulate prostacyclin release remains to be established. PMID- 3046957 TI - [The development of newborn C 57 BL/KsJ-dbm mice produced from eggs fertilized in vitro]. AB - The purpose of this study was to examine the development of newly born C57BL/KsJ dbm mice produced from eggs fertilized in vitro. The embryos derived from fertilization in vitro (which was performed by using db/db eggs and adrenalectomized db/db (Adrex) spermatozoa,) were transferred to the oviduct of MRL/MpJ pseudopregnant recipients 30 hr after insemination. 376 of these embryos yielded 65 young. Weight gain and urine glucose, plasma glucose and insulin levels were measured in these young as well as in Adrex males. The young produced by fertilization in vitro showed hyperglycemia, hyperinsulinemia and obesity. The physiological abnormalities in these young were similar to those in db/db young produced by natural mating between heterozygote (db/+) males and females. Adrex males did not show hyperglycemia but did show hyperinsulinemia. These results indicate that in vitro fertilization and embryo transfer is an effective means of producing fetuses or newborns with an overt genotype in genetically diabetic obese (db) mice. PMID- 3046959 TI - Localization of brain type creatine kinase in kidney epithelial cell subpopulations in rat. AB - Immunoperoxidase studies of rat kidney using antibody to brain type isoenzyme of creatine kinase (BB) revealed a specific staining in the epithelial cells of the thick ascending limb of the Henle's loop and collecting tubule. Occasional epithelial cells in cortical tubules that lack brush border were also positive for BB. Renal glomeruli and proximal tubules showed no immunoreactivity to this enzyme. PMID- 3046961 TI - Acquired immunodeficiency syndrome research in critical care: a review and future directions. PMID- 3046958 TI - The nutritional incidence of flavonoids: some physiological and metabolic considerations. AB - Examination of the physiological activity of flavonoids in relation to their antiscorbutic properties shows that some of these compounds, the flavan-3-ols, have a particular nutritional impact and consequently should be distinguished from the rest of the flavonoids and polyphenols. Therefore, the use of the term 'Vitamin P' and 'Bioflavonoids' is also discussed. PMID- 3046960 TI - Aldolase C is localized in neuroendocrine cells. AB - To elucidate the localization of the subunit C of aldolase (aldolase C) in peripheral neuroendocrine cells, we made an immunohistochemical study with monospecific antibodies against human aldolase C. Aldolase C was found to be localized in various types of neuroendocrine cells; in the pituitary gland, thyroid, pancreas, adrenal gland, bronchus, and gastrointestinal tract. PMID- 3046962 TI - Purification of meiotic spindles and cytoplasmic asters from mouse oocytes. AB - The unfertilized mouse oocyte is arrested at second metaphase of meiosis with microtubules existing exclusively in the meiotic spindle. Multiple inactive cytoplasmic microtubule organizing centers (MTOCs) are also present. These MTOCs can be identified immunocytochemically with an autoimmune serum (No. 5051) directed against pericentriolar material (PCM) and also by their nucleating capacity in the presence of taxol which effectively lowers the critical concentration for tubulin polymerization. Taxol induces the formation of cytoplasmic microtubule asters around the PCM foci, a process which also occurs in untreated eggs after fertilization. The molecular characterization of these structures has not been undertaken previously, probably due to the very small amount of material available. We have developed a single-step purification procedure by which very clean preparations of meiotic spindles and cytoplasmic asters can be obtained, as judged by phase-contrast microscopy and transmission electron microscopy. The purified structures were shown to correspond to those observed in vivo: positive staining of the spindles was observed with anti tubulin and anti-phosphoprotein (MPM2) antibodies, and positive staining of the MTOCs was observed with MPM2, No. 5051, and anti-calmodulin antibodies. As expected, tubulin was the major protein present in the preparations. Silver staining of SDS-PAGE also revealed the presence of a small number of other polypeptides (Mr of around 47, 35, and 25K). Amongst newly synthesized polypeptides associated with the preparation, two prominent high molecular weight proteins (greater than 200K) were enriched in addition to tubulin and polypeptides with Mr of around 52, 41, and 35K. PMID- 3046963 TI - A secretory protein restricted to type I cells in neonatal rat submandibular glands. AB - The perinatal submandibular gland of the rat contains an 89-kDa secretory protein (Protein C) that is released upon cholinergic stimulation. Polyclonal antibodies raised against Protein C show that this protein is localized in the Type I cells and is not found in typical Type III cells. However, morphological variants of Type III cells (Type IIIP) contain material that is cross-reactive with antibodies to Protein C. Cross-reactive components also are found in mucous cells of the neonatal sublingual glands, parotid and minor sublingual glands, and adult submandibular and sublingual glands. Immunoblots of electrophoretically separated proteins show a distinct Protein C band at 89 kDa only in neonatal submandibular glands; neonatal sublingual and minor sublingual glands show some diffuse reactivity over a range of mobilities encompassing that of Protein C. We propose that the cross-reactive components of mucous cells and Type IIIP cells are not Protein C, but different proteins associated with mucous differentiation, and that the Type IIIP cells of the neonatal submandibular gland are in transition from Type III to mature mucous cells. PMID- 3046964 TI - Descriptive and mechanistic considerations of interleukin 1 and insulin secretion. AB - Insulin-dependent diabetes mellitus (IDDM) may be mediated in part by an autoimmune mechanism, as suggested by associated cytologic and serologic phenomena, e.g., insulitis, beta-cell necrosis, and the presence of both islet cell and insulin antibodies. Immunological approaches to the prediction and intervention in the progression of beta-cell destruction in this disease are under evaluation. A recent hypothesis is that cytokines, including interleukin 1 (IL-1), play causative roles in such autoimmune processes. Several studies have convincingly demonstrated that IL-1 is a potent modulator of beta-cell function and can potentiate or inhibit glucose-induced insulin secretion, depending on the concentration and length of exposure to IL-1. IL-1 alone or in concert with other cytokines is cytotoxic to beta-cells. The cellular mechanisms responsible for the potent effects of IL-1 on the beta-cell are unknown and just beginning to emerge. Although speculative at this time, this perspective delineates cellular mechanisms that are likely to represent possible primary sites for the IL-1 action on beta-cells. A mechanistic understanding of the effects of IL-1 on the beta-cell may clarify its role in modulating insulin release in vivo or yield insight into the pathogenesis of IDDM. PMID- 3046965 TI - Rapid reduction and return of surface insulin receptors after exposure to brief pulses of insulin in perifused rat hepatocytes. AB - Hepatic receptors are normally exposed to discrete pulses of insulin and glucagon at intervals of 8 to 16 min. Using a multicolumn system for perifusing hepatocytes, we investigated the effect of this pattern on the normal processing of the insulin receptor. Surface-receptor binding was measured in acid-washed cells harvested from individual columns. The number of high-affinity surface receptors fell to a nadir 1 min after the end of a 3-min square-wave pulse of insulin. The maximum reduction reached 45% of baseline at amplitudes of 1000 microU/ml or above. The number of surface binding sites returned to baseline 15 min after the end of the pulse, but the affinity constant of the high-affinity receptor was unchanged. The reduction of surface binding was dose dependent, with an ED50 of 251 +/- 34 microU/ml. Prolonging the pulse to 60 min did not affect the nadir or the rate of restoration of the surface-receptor population. The change in surface binding was reduced at 15 degrees C and abolished at 4 degrees C. After a pulse, the pattern of change was a period of rapid decline to a nadir (t1/2 less than or equal to 1 min) that persisted for 3-5 min, followed by restoration of surface binding that reached baseline in 10-15 min. This same pattern was present after six ED95 pulses delivered at intervals of 15 min. These data indicate that the internalization of hepatocyte surface receptors and their recycling and reinsertion into the plasma membrane can be entrained to pulses at the physiologic pulse frequency. PMID- 3046966 TI - Insulin effects on pantothenic acid uptake in isolated perfused working hearts from diabetic rats. AB - Pantothenic acid uptake was studied in isolated working hearts from spontaneously diabetic BB Wistar and streptozocin-induced diabetic (STZ-D) rats. If insulin treatment was stopped for a 24-h period from spontaneously diabetic rats, a significant decrease in the rate of pantothenic uptake was noted (from 147.3 +/- 5.0 to 110.8 +/- 10.6 nmol.g-1 dry wt.30 min-1). Pantothenic acid uptake rates were also reduced in 48-h STZ-D rats (118.0 +/- 6.1 nmol.g-1 dry wt.30 min-1, compared to 158.2 +/- 5.3 in control rats). The decrease in pantothenic acid uptake in all diabetic animals occurred whether hearts were perfused with 1.2 mM palmitate or 1.2 mM palmitate and 11 mM glucose. If insulin (500 microU/ml) was added to the perfusion medium of hearts from spontaneously diabetic rats perfused with palmitate and glucose, a significant increase in pantothenic acid uptake was noted (from 110.8 +/- 10.6 to 167.0 +/- 9.4 nmol.g-1 dry wt.30 min-1). Insulin had no significant effect on pantothenic acid uptake in hearts from spontaneously diabetic rats perfused with palmitate alone. In STZ-D rats, insulin added to hearts perfused with palmitate and glucose resulted in a small but significant increase in pantothenic acid uptake (from 118.0 +/- 6.1 to 130.6 +/- 4.0 nmol.g-1 dry wt.30 min-1). Insulin had no effect on pantothenic acid uptake in control hearts perfused either in the presence or absence of glucose. These data suggest that insulin, in the presence of glucose, can increase pantothenic acid uptake in diabetic rats. PMID- 3046967 TI - Reduction of insulin clearance during hyperglycemic clamp. Dose-response study in normal humans. AB - Insulin secretion and clearance were studied in eight normal subjects who underwent hyperglycemic clamp studies at plasma glucose levels of 120, 225, and 300 mg/dl on three occasions. Insulin secretion rates were calculated during a 1 h baseline period and during 3 h of glucose clamping from a two-compartmental analysis of peripheral C-peptide concentrations with individual kinetic parameters derived after intravenous bolus injections of biosynthetic human C peptide. At the 300-mg/dl clamp level, the insulin secretion rate increased to a value 9.9 +/- 0.7 times that of basal at the end of the clamp (mean +/- SE), whereas over the same period, the peripheral insulin concentrations increased to a greater extent, reaching a value 15.4 +/- 1.2 times that of basal (P = .002). This greater relative increase in the insulin concentration in comparison with the corresponding insulin secretion rate suggests a reduction in the clearance of endogenous insulin. A similar trend was seen at the 225-mg/dl clamp level, but the relative increase in the insulin concentration (9.9 +/- 1.5 times that of basal) was not significantly higher than the relative increase in the insulin secretion rate (8.1 +/- 0.5 times that of basal, P = .17). At the 120-mg/dl clamp level, the relative increases in the insulin secretion rate (2.7 +/- 0.2 times that of basal) and the insulin concentration (2.4 +/- 0.2 times that of basal) were similar (P = .26), indicating no reduction in endogenous insulin clearance during moderate stimulation of insulin secretion. In conclusion, a reduction in endogenous insulin clearance occurs during greater stimulation of insulin secretion at higher glucose-clamp levels.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3046968 TI - Measurement of L-[1-14C]leucine kinetics in splanchnic and leg tissues in humans. Effect of amino acid infusion. AB - Although whole-body leucine flux is widely measured to study body protein turnover in humans, the contribution of specific tissues to the total-body measurement remains unknown. By combining the organ-balance technique with the systemic infusion of L-[1-14C]leucine, we quantitated leucine production and disposal by splanchnic and leg tissues and by the whole body, simultaneously, in six normal men before and during amino acid infusion. At steady state, disposal of arterial leucine by splanchnic and leg tissues was calculated from the percent extraction (E) of L-[1-14C]leucine counts: uptake = E x [Leu]a x flow. Tissue release of cold leucine (from protein turnover) into vein was calculated as the difference between leucine uptake and the net tissue leucine balance. In the postabsorptive state, despite substantial (P less than .01) extraction of L-[1 14C]leucine by splanchnic (23 +/- 1%) and leg (18 +/- 2%) tissues, net leucine balance across both tissue beds was small, indicating active simultaneous disposal and production of leucine at nearly equivalent rates. Splanchnic tissues accounted for approximately 50% of the measured total-body leucine flux. During amino acid infusion, the net leucine balance across splanchnic and leg tissues became positive, reflecting not only an increase in leucine uptake but also a marked suppression (by approximately 50%, P less than .02) of cold leucine release. This reduction in splanchnic and leg leucine release was indicated by a sharp decline in whole-body endogenous leucine flux.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3046969 TI - Activation of smooth muscle cell outgrowth from BB/Wor rat aortas. AB - An essential event in atherogenesis is the migration and proliferation of vascular smooth muscle cells (VSMCs), which contributes to the fibrocellular component of the atherosclerotic intimal thickening. We have been modeling this process by studying the outgrowth of VSMC from aortic explants onto tissue culture plastic substrate. Our hypothesis is that certain risk factors for atherosclerosis favor increases in subpopulations of cells with enhanced responsiveness to a variety of migratory and proliferative stimuli, in vivo and in vitro. As a known risk factor for atherosclerosis, diabetes might be reasonably postulated to induce such cell populations with altered susceptibility to additional noxious stimuli. We have tested this hypothesis with aortic explants from rats of the spontaneous autoimmune diabetic BB/Wor group, including diabetes-resistant, diabetes-prone, and treated acutely (less than 2 wk after onset of hyperglycemia) and chronically (greater than 2 wk) diabetic strains. As a group, the VSMCs from BB/Wor animals showed enhanced outgrowth in 0.1% serum (23-48%) compared with VSMCs from ordinary Wistar rats (less than 10%). Within the BB/Wor group, however, the rank order of enhanced outgrowth was diabetes resistant greater than chronically diabetic greater than acutely diabetic subjects. When the outgrowth assay was performed in the presence of supplemental insulin (10 mU/ml), outgrowth was increased, especially for the diabetes-prone animals, giving a new rank order of diabetes-prone greater than acutely diabetic greater than diabetes-resistant greater than chronically diabetic subjects. These data suggest a genetic predilection in the entire BB/Wor rat group for increased VSMC responsiveness to migratory and proliferative stimuli, a sensitivity of these VSMCs to insulin, and the dissociation of this vascular cell effect from other manifestations of diabetes, e.g., hyperglycemia. PMID- 3046970 TI - Insulin autoantibodies and insulin assay. AB - Insulin antibodies are known to interfere with the radioimmunoassay of insulin. We tested intravenous glucose tolerance on 25 insulin autoantibody-positive (IAA+) patients and 25 IAA- controls, who were matched for sex, age, and body mass index, to establish if IAA could also interfere with insulin assay. Insulin content was measured in untreated serum, serum precipitated with polyethylene glycol (PEG, free insulin), and serum extracted with acid and precipitated with PEG (total insulin). The mean untreated first-phase insulin response (I1 + 3) for IAA+ patients was 172 +/- 67.3 mU/L, significantly higher than the mean control value of 108 +/- 47.5 (P less than .001). After PEG precipitation, mean I1 + 3 in the patient group fell significantly to 105 +/- 48.4 mU/L (P less than .001), but the control value was unchanged (104 +/- 45.5). The mean percentage fall after PEG precipitation was 36.9% (patients) and 2.9% (controls) (t = 8.3, P less than .001). There was a strong correlation between the IAA titer and the interference in the insulin assay (r = .81). After total insulin extraction of IAA samples, there was a significant fall in mean I1 + 3 to 134 +/- 55.4 mU/L (P less than .001), but the control value was unchanged. IAA can significantly falsify insulin measurement, and care must be taken in the interpretation of insulin-release tests when IAA is present. PMID- 3046971 TI - IDDM in BB rats. Enhanced MHC class I heavy-chain gene expression in pancreatic islets. AB - Modulation in major histocompatibility complex (MHC) gene expression correlates with the inflammatory reactions that occur during graft rejection and autoimmune disease. We analyzed the expression of class I and II MHC genes in the pancreatic islets of prediabetic and newly diabetic BB rats by immunohistochemistry of tissue sections and Northern blotting of RNA extracted from isolated islets. We show that enhanced levels of MHC class I heavy-chain RNA are present in pancreatic islets before overt inflammation and the onset of insulin-dependent diabetes mellitus (IDDM) in the spontaneously diabetic BB rat. Immunohistochemical analysis revealed enhanced class I antigen expression throughout the pancreatic islets of newly diabetic animals but no induction of class II antigen on endocrine cells within the islet. Varying degrees of inflammatory infiltrate were observed in the sections exhibiting enhanced class I antigen expression or in nearby serial sections. Southern blot analysis revealed no restriction-fragment-length polymorphism or amplification of the endogenous class I heavy-chain genes compared with those of seroidentical disease-resistant Wistar-Furth rats. I-A alpha and I-E alpha hybridizing RNA appeared de novo before overt diabetes, although concomitantly with T-lymphocyte-receptor beta chain and interferon-gamma gene hybridizing RNA and after MHC class I heavy-chain RNA enhancement was observed. These data indicate the possibility that enhanced class I heavy-chain gene expression plays a role in the progression of IDDM. PMID- 3046972 TI - Glucose-regulated proinsulin processing in isolated islets from rat pancreas. AB - We demonstrated previously that the conversion rate of proinsulin to insulin in pancreatic islets progressively increased after prolonged prior exposure to glucose (11 mM) and that this effect could be blocked by cycloheximide. This study was designed to characterize further the time course and regulation of the proinsulin conversion process. The effects of prior exposure to glucose on proinsulin conversion were dose dependent (Km, approximately 7 mM glucose) and time dependent, taking approximately 3 h to reach the maximum rate. Glucose added at or after the subsequent [3H]leucine pulse was ineffective. Mannoheptulose, added during a 3-h exposure with glucose (11 mM), prevented glucose-induced activation of the proinsulin conversion process. L-Leucine (20 mM) was as effective as 11 mM glucose in activating conversion, whereas 2-alpha ketoisocaproic acid (20 mM) or phorbol ester (50 nM) had little effect. Activation of proinsulin conversion by a 24-h exposure to glucose (11 mM) was reversed by a subsequent 3-h prior exposure to cycloheximide. alpha-Amanitin, an inhibitor of mRNA synthesis, did not influence the glucose-induced activation of proinsulin conversion when present during a 3-h exposure to glucose; however, it completely inhibited glucose-stimulated conversion when present during 24 h exposure. Results suggest that activation of the proinsulin conversion process is regulated by glucose metabolism rather than the glucose molecule per se and that other, but not all, secretagogues are effective. Conversion may require prior synthesis of a pool of converting enzyme(s) or other regulatory proteins whose turnover is relatively rapid (approximately 33 h) and whose mRNA is more stable (to 24 h). PMID- 3046973 TI - Stimulatory effects of cholecystokinin on isolated perifused islets inhibited by potent and specific antagonist L 364718 [corrected]. AB - The influence of L 364718 on islet responsiveness to sulfated cholecystokinin (CCK-8S) was investigated. In islets whose inositol-containing phospholipids were prelabeled during a 2-h incubation period, subsequent exposure to L 364718 (1 nM) significantly impaired the secretion of insulin usually noted in response to 200 nM CCK-8S in the simultaneous presence of 7 mM glucose. A higher level of the antagonist (10 nM) completely abolished insulin secretion. L 364718 (1-10 nM) reduced the efflux of 3H from myo-[2-3H]-inositol prelabeled islets in parallel with the reduction in secretion. L 364718 (10 nM) significantly reduced the accumulation of 3H-containing inositol phosphates usually noted with CCK-8S addition. L 364718, at levels 10- to 100-fold greater than those necessary to attenuate CCK-8S-induced insulin secretion, had no adverse effect on the insulin secretory response of freshly isolated islets to 10 mM glucose alone, 5 mM D glyceraldehyde, 15 mM alpha-ketoisocaproate, or 50 ng/ml gastric inhibitory polypeptide. L 364718 (1000 nM) had no adverse influence on carbamylcholine (1 mM)-induced phosphoinositide hydrolysis. These results establish L 364718 as a potent and highly selective antagonist of cholecystokinin's stimulatory actions on beta-cells. Because of its potency, selectivity, and oral effectiveness, in vivo studies with L 364718, aimed at unraveling the pleiotropic effects of CCK-8S on glucose and insulin homeostasis, seem feasible. PMID- 3046974 TI - Albert E. Renold: July 10, 1923-March 21, 1988. PMID- 3046975 TI - Alloxan: history and mechanism of action. PMID- 3046976 TI - Serum proinsulin levels at fasting and after oral glucose load in patients with type 2 (non-insulin-dependent) diabetes mellitus. AB - A simple and sensitive human proinsulin radioimmunoassay system was developed using guinea pig anti-proinsulin serum, which cross-reacted neither with human insulin nor C-peptide. The recognition site of the antiserum seems to be located near the junction between the B chain and C-peptide. With this assay system, we studied the serum proinsulin concentration at fasting and after an oral 100 g glucose load in 25 healthy subjects, 21 subjects with impaired glucose tolerance and 40 patients with Type 2 (non-insulin-dependent) diabetes mellitus. At fasting, serum proinsulin was 5.8 +/- 3.3 pmol/l in normal subjects as compared to 9.5 +/- 6.9 pmol/l (p less than 0.05) in subjects with impaired glucose tolerance and 12.6 +/- 7.5 pmol/l (p less than 0.001) in diabetic patients. The molar ratio of proinsulin to insulin was also increased in subjects with impaired glucose tolerance or diabetes compared to control subjects. After a 100 g oral glucose load, serum proinsulin increased more slowly than insulin. The proinsulin response after an oral glucose load was augmented in subjects with impaired glucose tolerance and diabetes, while the insulin response decreased with the elevation of fasting plasma glucose. Diabetic patients with high fasting plasma glucose had a very poor insulin response, but the proinsulin response was similar to control subjects. There was a linear correlation between summed proinsulin values and summed insulin values, but the slope of the regression line was steeper in diabetic patients than in control subjects. There was a relative increase in serum proinsulin both in subjects with impaired glucose tolerance and diabetic patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3046977 TI - Effect of haemolysis on radioimmunoassay for insulin. PMID- 3046978 TI - Is insulin atherogenic? PMID- 3046979 TI - Effects of thyroxine-driven precocious metamorphosis on maturation of adult-type allograft rejection responses in early thyroidectomized frogs. AB - In contrast to the adult pattern of allograft reactivity, larval South African clawed frogs (Xenopus) either become tolerant of adult major histocompatibility complex (MHC) disparate skin grafts or reject them slowly. Larvae fail to reject grafts that are incompatible at minor histocompatibility (H) loci (MHC identical); rather, they become immunologically tolerant. We report here that early thyroidectomized (thyroidx) larvae, forced to metamorphose precociously on a regimen of thyroxine (T4) treatment, showed larva-like responses in that their ability to reject skin allografts that differed by minor H loci or one MHC haplotype was significantly impaired relative to age-matched intact postmetamorphic controls. During the normal ontogeny of Xenopus, lymphocyte numbers increase in larval life, decrease during metamorphosis, and increase to adult levels at 6-12 months of age. Thyroidectomy and a low-dose regimen of thyroxine treatment limited the number of lymphocytes that developed in larval organs and very dramatically reduced those populations below control levels during metamorphosis. Because these changes were stage-dependent rather than age dependent, the second wave of lymphopoiesis in thyroidx frogs treated with low T4 may occur out of synchrony with the expression of Class I MHC antigens. Impaired allograft rejection, therefore, may reflect the absence of a population of cells that can effectively recognize minor H alloantigens and/or self and allo Class I antigens. PMID- 3046980 TI - [Guided biopsy of tumors of the liver: a consensus should be hoped?]. PMID- 3046981 TI - [Puncture of solid tumors of the liver with large caliber needle guided by ultrasonography. Study of 70 punctures]. AB - Seventy specimens of solid hepatic tumors were obtained for histologic examination with a 18 G (1.2 mm) diameter needle under real-time ultrasound control. Overall sensitivity was 95.6 p. 100 while specificity was 100 p. 100. Distinction between primitive and secondary malignant tumor was possible in 91 p. 100 of cases. Transient hemorrhage at the puncture site occurred in one patient. These results attest to the superiority of guided puncture with a needle of sufficient caliber, thus allowing for correct histologic study. Side effects did not occur more frequently than with fine caliber needles. PMID- 3046982 TI - [Morphological diagnosis of pseudocystic tumors of the liver]. AB - Ten cases of pseudocystic hepatic tumors were observed during the past ten years. This entity is very rare in comparison with benign hepatic cysts. Careful analysis of the characteristics of these lesions usually allows to differentiate them from benign cysts. Of importance are wall thickness, mural nodules and fluid/fluid levels on ultrasound and/or computed tomography scanning. In our series, sonography depicted the true morphology of these cystic masses more clearly than computed tomography. If definitive diagnosis cannot be made by morphologic examinations, percutaneous aspiration of the cystic lesion, a simple and safe method, should be considered. PMID- 3046983 TI - [Treatment of alcoholic hepatitis. A review of randomized trials]. PMID- 3046984 TI - [Plasma insulin and glucagon after ingestion of meal enriched with increased doses of pectin in healthy man]. AB - The effects of three doses of pectin (5, 10 and 15 g) included in a solid-liquid meal on the postprandial plasma insulin and glucagon responses were studied in 12 healthy men. The mean plasma glucagon level was significantly smaller with 5 g of pectin than the control values at 150 min (p less than 0.05) whereas plasma insulin values did not vary. No change in mean plasma glucagon and insulin levels was noted with 10 g and 15 g of pectin although the mean blood glucose levels were significantly higher than the control values at 180 min (p less than 0.05). Addition of pectin to a meal, even if the doses were relatively important, had little or no effect on the postprandial hormonal responses in healthy men. However, pectin could be of renewed interest because of the possibility of its action of satiety by means of sustained late blood glucose levels. PMID- 3046985 TI - [Culture of gastroenteropancreatic endocrine tumors: current contributions and future prospects]. PMID- 3046986 TI - [Hepatitis caused by cyproheptadine (Periactine). A case and review of the literature]. AB - We report the case of a patient who developed jaundice after receiving cyproheptadine for 29 days. Complete recovery occurred within 3 months after cyproheptadine withdrawal. Analysis of 3 previously reported cases and this observation shows that cyproheptadine-induced hepatitis is uncommon. Hepatitis occurs within the first month of treatment and is of the cytolytic or mixed pattern type. Jaundice is constant. Most often, recovery occurs quickly after the discontinuation of cyproheptadine. However, acute hepatitis can be followed by prolonged anicteric cholestasis. PMID- 3046987 TI - [Neuroma of the common bile duct: a rare cause of jaundice]. AB - We report a case of neuroma of the main bile duct arising twenty years after cholecystectomy. The patient, a 82-year-old woman, was admitted for jaundice. Endoscopic retrograde cholangiography showed a regular stenosis of the main bile duct. Histologic examination demonstrated neuroma. Based on the analysis of this and 15 other previously published cases, the following features of bile duct neuroma were outlined: a) variable interval between cholecystectomy and the onset of jaundice (6 months to 35 years); b) the generally complicated postoperative course, c) the various localizations on the biliary tree (cystic, main bile duct, intrahepatic bile duct) and, d) the circumstances of onset. PMID- 3046988 TI - [Natural history of glandulocystic fundic polyposis in Gardner's syndrome]. AB - The authors report 3 cases of fundic gland polyposis associated with Gardner's syndrome. Control endoscopies revealed fundic gland polyposis disappearance 14, 14, and 7 years respectively after discovery, and colectomy performed at ages 29, 30, and 32, respectively. This regression, described in autonomous fundic gland polyposis, is poorly documented in Gardner's syndrome. This phenomenon is not related to colectomy. However no explanation is currently available, and further studies are necessary to determine the factors related to the natural history of fundic gland polyposis in familial adenomatosis coli/Gardner's syndrome. PMID- 3046989 TI - [Topographic-anatomic considerations in vaginal surgical therapy of stress incontinence]. AB - The medial pubourethral ligament and the posterior pubourethral ligaments have the important task (for continence) of fixing the neck of the bladder. If the neck of the bladder is placed in relationship to height of the posterior wall of the symphisis, viscerographic examinations show that the neck of the bladder is high in 9% of cases, medium-high in 26%, and low in 65%. When performing reconstructive surgery following detachment of the neck of the bladder as a result of abdominal pressure, these topographic-anatomic features can only be taken into consideration if the primary, original site of the neck of the bladder is known. Intraoperative exposure of the ligament insertion and of the posterior pubourethral ligaments will always permit the primary location of the neck of the bladder to be determined. This is situated 2.3 cm beneath the insertion, and joining the medial ligament to the dorsal urethra will guarantee normal repositioning of the neck of the bladder. If the primary location of the neck of the bladder is unknown in the individual case being treated, the deep tissue pad which develops subsequent to anterior colporraphy corresponds to a nonspecific tissue accumulation with incalculable outcome. PMID- 3046990 TI - [Urodynamic and roentgenologic changes following ventral levatorplasty]. AB - Our first 23 patients treated by this method were followed up by urodynamic assessment and urethrocystovaginography 13 months after the operation. Urodynamic assessment was performed according to the criteria of the International Continence Society. There was no statistically significant difference in the resting urethral closure pressure (UCP), neither at maximum point nor at 30% or 70% of the urethral functional length (UFL). The postoperative reduction of UFL was statistically significant but did not exercise any significant influence on continence. The maximum bladder volume was not affected. For assessing the urethral stress profile we calculated the depression quotient which was significantly unfavourable at 30% of UFL but not affected at 70%. The lateral urethrocystovaginogram was interpreted according to Green. Most of the parameters were significantly improved. At the time of reexamination 70% of the patients were subjectively continent, whereas in 21.7% the incontinence remained unchanged and deteriorated in 8.6%. In our opinion, this modified anterior repair is not recommended as a method to encourage a wider range of indications for the vaginal repair of incontinence. Hence, this operation method should be performed only in case of an evident insufficiency of the vesicourethrovaginal septum. PMID- 3046991 TI - [Jejunal atresia: contribution to prenatal ultrasonic diagnosis]. AB - In a case report we present the prenatal diagnosis of an intestinal obstruction. The problem of locating exactly the area of obstruction is discussed. Prenatal diagnosis permits optimal management after birth and can lead to a better prognosis. PMID- 3046992 TI - [Cholaemic placentosis--a contribution to metabolic disorders of the placenta]. AB - The deposition of bile substances produces intracellular metabolic disorders in the organs so affected. Since severe cholaemia is rare in pregnancy, similar disorders of the placental cells have been not discussed up to now. We report on morphological observations in a placenta of a stillbirth in the middle of pregnancy. Bile substances are deposited in the syncytiotrophoblast and Hofbauer cells, pointing to the probable function of the Hofbauer cells as macrophages. PMID- 3046993 TI - Probucol in long-term treatment of hypercholesterolemia. AB - Over 15 yr of clinical experience with probucol seems to indicate that the drug is a safe and moderately effective hypocholesterolemic drug even in long-term treatment. Adverse effects are infrequent and usually tolerable. Probucol prolongs QT-interval but this does not seem to be connected with harmful effects in man. Its hypocholesterolemic action is not sufficient in all patients but this can be substantially improved by combining other hypolipidemic drugs with different mode of action. Interference with lipoprotein structure may contribute to the hypocholesterolemic action of probucol, and despite the lowering of HDL cholesterol probucol has been shown to regress peripheral cholesterol deposits. There is also tentative evidence that the drug may protect from CHD in primary prevention. The newly-discovered antioxidant property of probucol preventing harmful LDL modification in vitro might partly explain the favorable effects of probucol independently of its effects on LDL- or HDL-cholesterol levels. PMID- 3046994 TI - Physiological and pharmacological regulation of small intestinal cholesterol synthesis. AB - 1. The small intestine is an important site of cholesterol synthesis in the body and at least in experimental animals, it also contributes to the circulating plasma pool of cholesterol. 2. Studies on synthesis regulation have been partly contradictory but it is now concluded that the cellular cholesterol balance is the basic regulatory factor of intestinal cholesterol synthesis. However, the balance is affected differently in various specialized cells and parts of the small intestine. 3. Most data on synthesis regulation are derived from experimental animals but the few human studies suggest that similar regulatory factors function in man, too. PMID- 3046995 TI - Speculations on the molecular nature of anesthesia. PMID- 3046996 TI - Drug distribution in the body. PMID- 3046997 TI - Pulmonary surfactant-associated proteins. PMID- 3046998 TI - Dry socket: prevention and treatment. PMID- 3046999 TI - Using a cast post and core to save an existing crown. PMID- 3047001 TI - Medical psychotherapy. Clinical application in an inpatient setting. AB - Under the prevailing Current Procedural Terminology (CPT) billing codes, the function of the inpatient psychiatrist is defined in terms of providing "psychotherapy." Various third-party auditors have raised the implication of fraud when this has not been provided in a standardized fashion. In response, revisions in the CPT codes have been proposed aimed at legitimizing the true responsibilities of the attending psychiatrist. As an alternative solution, I have attempted to briefly summarize the concept of "medical psychotherapy," clarify how this approach would apply in an acute inpatient setting, and outline how it differs from the more classically defined outpatient psychotherapy models. PMID- 3047000 TI - Measurement of plasma renin activity in the freshwater turtle. AB - Components of the renin angiotensin system have been identified in many nonmammalian vertebrates. However, in many of these animals, including reptiles, the physiological functions and importance of the system remain unclear. To aid in the study of the system in a reptile we modified a commercially available radioimmunoassay (RIA) kit containing antibody against human angiotensin I (ANG I) for use in the freshwater turtle, Pseudemys scripta. Cross-reactivity between anti-human ANG I antibodies (Rainen Angiotensin I RIA Kit, New England Nuclear) and turtle ANG I was demonstrated. Cross-reactivity with the antibody in two other human ANG I assay kits (Travenol-Genentech and Biotecx) was very limited. Blood for assay was collected from conscious turtles in EDTA, centrifuged, and the plasma frozen at -20 degrees. Turtle ANG I was generated by incubation at 0.5 ml plasma at pH 5.5 for 2 hr at 30 degrees with addition of dimercaprol and 8 hydroxyquinoline. Angiotensin generation increased with temperature and with generation time. The recovery of turtle ANG I added to turtle plasma prior to incubation was 92-97%. The assay procedure was used to measure plasma renin activity (ng/ml/hr incubation) from unstimulated turtles. PMID- 3047002 TI - [Cloning of the gene of Saccharomyces cerevisiae yeasts that determines resistance to the toxic action of cadmium ions]. AB - A gene conferring resistance to cadmium in Saccharomyces cerevisiae was isolated from a yeast gene library created on the basis of the pL3 vector. The phenotype of resistance is only expressed in the yeast cells with cloned DNA inserted into a multicopy plasmid. Integration of the plasmid into chromosome or introduction of the centromeric region into the plasmid decreases the level of cadmium resistance. The cloned Sau3A I fragment of the yeast chromosome is 3.5 kbp in size. Restriction analysis and subcloning experiments showed the gene to be located within 1.6 kbp of the XhoI-Sau3A I fragment of DNA. Instability was observed in the vicinity of the XhoI-Sau3A I fragment of the yeast DNA in Escherichia coli. PMID- 3047003 TI - [The role of the RS1 sequence of Vibrio cholerae in the amplification of a segment of plasmid DNA carrying the tetracycline resistance gene and cholera toxin genes]. AB - The clones with 4 to 30-fold increased level of tetracycline resistance (TcR) were selected from the strain of Escherichia coli K-12 carrying the pCO107 plasmid. The plasmid is the cointegrate formed from the plasmids pOX38 (F derivative) and pCT105 (pBR322-derivative carrying the vct operon of Vibrio cholerae eltor and the RSI sequence). pCO107 contains RSI at the junctions of two plasmid genomes. Using restriction analysis and Southern blot hybridization technique it was shown that increased tetracycline resistance is accompanied by amplification of the pCT105 segment of pCO107 and depends upon the presence of direct repeats of flanking RSI. Amplification of the pCT105 also resulted in increased production of the cholerae toxin (CT). PMID- 3047004 TI - [Localization of the multigene Lpm locus in chromosome 9 of the American mink]. AB - The results of genetic study on linkage of Lpm locus with peptidase B gene are presented. Investigation of 111 offspring back-crosses shows that Lpm allotypes and allelic variants of peptidase B are inherited in concert. The frequency of recombination between the Lpm locus and peptidase B gene is 11 +/- 3% in male. Since it was earlier established that peptidase B gene is a marker of chromosome 9, our data indicate that the Lpm loci family is situated in the chromosome 9 of domestic mink. PMID- 3047005 TI - [A method of analyzing the chromosome localization of repetitive nucleotide sequences by using hybridization in situ]. AB - To increase precision in the quantitative analysis of chromosomal distribution of repeated DNA sequences detected by in situ hybridization, a number of factors important in studies of the molecular basis of chromosome polymorphism should be considered. While analysing results of hybridization, one should only use the number of grains over the sites of their regular localization, instead of those over the whole chromosome. It is also evident that to decrease variability conditioned by differences in the labelling degree of separate cells, the relative numbers of labelled grains over chromosomes should be used in the analysis and not the absolute ones. Data from those cells ought to be only included in the analysis, in which the labelling degree is sufficient to show all localization sites of the repeated sequences studied, i.e. cells should be used with comparatively constant relative numbers of labelled grains over all their regular localization sites. PMID- 3047006 TI - Differential inhibition of the initiation of DNA replication in stringent and relaxed strains of Escherichia coli. PMID- 3047007 TI - Ribosomal protein synthesis is not regulated at the translational level in Saccharomyces cerevisiae: balanced accumulation of ribosomal proteins L16 and rp59 is mediated by turnover of excess protein. AB - We have investigated the mechanisms whereby equimolar quantities of ribosomal proteins accumulate and assemble into ribosomes of the yeast Saccharomyces cerevisiae. Extra copies of the cry1 or RPL16 genes encoding ribosomal proteins rp59 or L16 were introduced into yeast by transformation. Excess cry1 or RPL16 mRNA accumulated in polyribosomes in these cells and was translated at wild-type rates into rp59 or L16 proteins. These excess proteins were degraded until their levels reached those of other ribosomal proteins. Identical results were obtained when the transcription of RPL16A was rapidly induced using GAL1-RPL16A promoter fusions, including a construct in which the entire RPL16A 5'-noncoding region was replaced with the GAL1 leader sequence. Our results indicate that posttranscriptional expression of the cry1 and RPL16 genes is regulated by turnover of excess proteins rather than autogenous regulation of mRNA splicing or translation. The turnover of excess rp59 or L16 is not affected directly by mutations that inactivate vacuolar hydrolases. PMID- 3047009 TI - Nucleotide sequence of the SUP2 (SUP35) gene of Saccharomyces cerevisiae. AB - A nucleotide sequence of the yeast Saccharomyces cerevisiae omnipotent suppressor SUP2 (SUP35) gene is presented. The sequence contains a single open reading frame (ORF) of 2055 bp, which may encode a 76.5-kDa protein. A single transcript of 2.3 kb corresponding to a complete ORF is found. Analysis of codon bias suggests that the SUP2 gene is not highly expressed. The C-terminal part of the deduced amino acid sequence shows a high homology to yeast elongation factor EF-1 alpha, whereas the N-terminal part is unique for the SUP2 protein. The N terminus contains a number of short repeating elements and possesses an unusual amino acid composition. Analysis of the nucleotide and deduced amino acid sequences indicates that three additional proteins could possibly be expressed, two of which might be initiated on internal ATG codons and a third might be formed by alternative splicing. One of these proteins is supposed to be imported into mitochondria. Possible functions of the SUP2 gene product(s), especially its putative activity as a soluble factor controlling the fidelity of translation, are discussed. PMID- 3047008 TI - Transcription by RNA polymerase II induces changes of DNA topology in yeast. AB - We show that induction of transcription of a CYC1-lacZ fusion gene, borne on a yeast plasmid, causes an increase in negative superhelicity of approximately five turns. This increase is abolished by deletion of either essential element of the CYC1 promoter, the upstream activation site (UAS), or the TATA boxes. Several experiments indicate that the size of the increase is proportional to the size of the transcribed region. First, an internal deletion removing half of the CYC1 lacZ transcribed region results in a plasmid whose negative superhelicity on induction is intermediate between promoter-deletion plasmids and the parental plasmid. Second, plasmids bearing insertions of a fragment containing the putative CYC1 terminator into the CYC1-lacZ fusion gene have relative negative superhelicities proportional to the length of the truncated fusion transcripts generated. A plausible model explaining these observations is that local unwinding of the double helix by transcribing RNA polymerase generates positively supercoiled DNA, which is subsequently relaxed by a topoisomerase. PMID- 3047010 TI - cDNA cloning and sequence analysis of a chicken gene expressed during the gonadal development and homologous to mammalian cytochrome P-450c17. AB - A cDNA clone, pLOA0511, was isolated from the cDNA library prepared from the left ovaries of one- to three-day-old chickens. The transcript corresponding to this cDNA clone is approx. 1.9 kb in length and is present in steroidogenic tissues (ovary, testis and adrenal gland) but is undetectable in non-steroidogenic tissues. Relative abundance of the transcript in the ovary and testis is high and developmentally regulated but is much lower in the adrenal gland. Relative levels of this transcript in the developing left ovary and the regressing right ovary of the one- to three-day-old chicken are similar. The nucleotide sequence of this cDNA clone shows significant homology to some mammalian cytochromes P-450. Out of the 508 amino acids (aa) coded, 244 and 243 aa residues are identical with those of the bovine and human cytochrome P-450c17 (steroid 17 alpha-hydroxylase/17,20 lyase), respectively. There are three highly conserved regions among the bovine, human and chicken sequences, one of which is unique to P-450c17. These data suggest strongly that this cDNA corresponds to the mRNA of a chicken cytochrome P 450c17. PMID- 3047011 TI - Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. AB - Plasmid expression vectors have been constructed that direct the synthesis of foreign polypeptides in Escherichia coli as fusions with the C terminus of Sj26, a 26-kDa glutathione S-transferase (GST; EC 2.5.1.18) encoded by the parasitic helminth Schistosoma japonicum. In the majority of cases, fusion proteins are soluble in aqueous solutions and can be purified from crude bacterial lysates under non-denaturing conditions by affinity chromatography on immobilised glutathione. Using batch wash procedures several fusion proteins can be purified in parallel in under 2 h with yields of up to 15 micrograms protein/ml of culture. The vectors have been engineered so that the GST carrier can be cleaved from fusion proteins by digestion with site-specific proteases such as thrombin or blood coagulation factor Xa, following which, the carrier and any uncleaved fusion protein can be removed by absorption on glutathione-agarose. This system has been used successfully for the expression and purification of more than 30 different eukaryotic polypeptides. PMID- 3047012 TI - MRI vs CT for diagnosing brain disorders in the elderly. AB - Following a very brief, simplified explanation of magnetic resonance imaging principles, the general advantages inherent in such a system, as compared to CT, are explored. Clinical exploitation of MRI's attributes are then specifically discussed and illustrated in multiple types of intracranial pathology. PMID- 3047013 TI - Pneumonia in the elderly: a review. AB - Elderly persons are at increased risk of acquiring pneumonia. Morbidity and mortality rates from pneumonia are also higher. Clinical manifestations of pneumonia in the elderly are frequently very subtle; however, a high index of suspicion will induce the physician to obtain a chest roentgenogram. A new lung infiltrate will confirm the diagnosis. Although the Gram's stain of a sputum smear is most helpful in the initial selection of a specific antibiotic, a good quality sputum is not usually available in the elderly, and empiric therapy reduces the morbidity and mortality from pneumonia. PMID- 3047015 TI - Why do we live so long? PMID- 3047014 TI - Late-onset rheumatoid arthritis vs polymyalgia rheumatica: making the diagnosis. AB - Differentiating polymyalgia rheumatica from the onset of rheumatoid arthritis in the elderly has been the cause of much unnecessary confusion. Differential diagnosis of these disorders can be straightforward. A strategy is outlined, comprising a complete history, attention to clinical signs, and appropriate use of laboratory diagnostics. The clinical picture of each disorder is discussed, as are common obstacles to diagnosis. PMID- 3047016 TI - The role of free radicals in leaf senescence. AB - Decreased catalase activity is a consistent feature of leaf senescence. Although not as well documented, superoxide dismutase appears generally to decrease during leaf senescence. These changes suggest that free radical levels are likely to be higher in senescing tissues. The hydrogen peroxide-scavenging ability of chloroplasts due to the activity of the enzymes ascorbate peroxidase, dehydroascorbate reductase and glutathione reductase appears to be established although there is no information on changes in levels of these enzymes in response to leaf senescence. In plants, unlike mammals, the direct reaction of glutathione with H2O2, catalysed by glutathione peroxidase, appears to be only a minor means of scavenging hydrogen peroxide. Senescence appears to be correlated with increases in lipid peroxidation and membrane permeability. The findings reviewed in this paper lend general support to the view that free radicals play a significant role in the multifactorial syndrome which constitutes leaf senescence. PMID- 3047017 TI - [Experimental study of the methods of combined administration of metals on their toxicokinetics in the body]. PMID- 3047018 TI - [M.V. Lomonosov's ideas concerning work safety and occupational hygiene of miners (on the 275th anniversary of his birth)]. PMID- 3047019 TI - [A.A. Letavet, the founder of occupational radiation hygiene]. PMID- 3047020 TI - Platelet contribution to cancer cell growth and migration: the role of platelet growth factors. AB - Blood platelets contain several growth factors and inhibitors. The better known among them are named platelet-derived growth factor (PDGF) and transforming growth factor beta (TGF beta), active as modulators of growth of normal mesenchymal and epithelial cells. Cancer cell growth in vitro seems to be independent of the effects of exogenous peptides, but dependent on the cell ability to release autocrine growth factors, similar to PDGF or TGF beta. In addition, platelet-associated growth factors and inhibitors, which are able to induce a fibrotic response in connective tissue cells, might also play a role to modulate the desmoplastic reaction surrounding the tumor. PMID- 3047021 TI - Platelet contribution to the formation of metastatic foci: the role of cancer cell-induced platelet activation. AB - Platelets are thought to be involved in the development of blood borne metastasis. Ultrastructural and experimental studies demonstrate that association between tumor cells and platelets with subsequent activation of the coagulation cascade takes place in malignancy. Several hypotheses have been proposed to explain the mechanisms by which tumor cells activate platelets including generation of thrombin, ADP release and involvement of arachidonate metabolism. Perfusion studies with human homologous systems showed that intact tumor cells and tumor cell microvesicles were able to induce platelet thrombogenicity under defined flow conditions. The presence of divalent cations and plasma factors was necessary for the cancer cells to exert their activating capacity. These results suggest a role for platelets in the development of secondary metastasis as well as in the thrombotic events of malignancy. PMID- 3047023 TI - Different expression of procoagulant activity in macrophages associated with experimental and human tumors. AB - Mononuclear phagocytes, an integral part of the lymphoreticular infiltrate of human and experimental tumors, might contribute to fibrin deposition within malignant tissues through the production of procoagulant activity (PCA). Recent studies in different types of experimental tumors and in some cancer patients indicate that the functional status in terms of PCA of mononuclear phagocytes at 'peripheral' sites is not the same as that of macrophages at the tumor site (tumor-associated macrophages, TAM). Peripheral blood monocytes and/or peritoneal macrophages from V2-carcinoma-bearing rabbits and from mice bearing different types of tumors, like cells from normal animals, have low levels of basal PCA and are able to respond with a marked increase in PCA to in vitro stimulation. Likewise, in patients with primary lung cancer, pulmonary alveolar macrophages (PAM) obtained from the contralateral side of the neoplasm and blood monocytes behave essentially as cells from normal individuals. In contrast, profound changes in PCA have been found in TAM. In some experimental tumors and in a number of the patients, macrophages taken at the tumor site express high levels of basal PCA, which most probably reflect in vivo activation. Local activation of macrophages for PCA production might contribute to fibrin deposition at the tumor site. In the other experimental tumors studied and in the remaining patients, not only have macrophages low basal PCA but they also fail to respond to various stimulants in vitro. Although the mechanisms underlying these changes and their pathophysiological significance remain to be established, it is apparent that neoplastic growth may modulate the expression of macrophage PCA at the tumor site.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3047022 TI - Role of the coagulation system in tumor-cell-induced platelet aggregation and metastasis. AB - The ability of tumor cells to initiate coagulation and subsequent platelet aggregation is believed to facilitate the metastatic process. The mechanism by which tumor cells initiate thrombotic alterations is unclear. We have purified a plasma membrane protein platelet aggregating activity/procoagulant activity (PAA/PCA) from several rodent tumors which initiates the coagulation of homologous plasma and aggregation of homologous platelets by a mechanism independent of factor VII. This protein does not possess any proteinase activity; however, its activity is dependent upon the presence of factor X. In addition, PAA/PCA requires reconstitution with phospholipid for expression of activity. These results suggest that tumor cells express a unique protein which possesses procoagulant activity resulting in thrombin generation. Thrombin is responsible for subsequent tumor-cell-induced platelet aggregation. PMID- 3047024 TI - Monocyte procoagulant activity as a peripheral marker of clotting activation in cancer patients. AB - Peripheral blood monocytes generate the procoagulant tissue factor in vitro and in vivo in response to stimulation by a variety of agents. Monocytes from cancer patients generate significantly increased tissue factor and a quantitative relationship exists between the levels of monocyte tissue factor (MTF) and levels of circulating fibrinopeptide A (FPA), a marker of in vivo clotting activation. Furthermore, monocytes from cancer patients have a greater procoagulant response to stimulation by endotoxin in vitro, which appears independent of lymphocyte regulation. These findings suggest a priming process in vivo, and may reflect exposure of monocytes to tumor antigen(s) or components of the immune response to tumors. PMID- 3047025 TI - Expression of plasminogen activator as a marker of stimulation in tumor associated macrophages. AB - Tumor-associated macrophages (TAM) are a peculiar subpopulation of resident phagocytes with activities possibly important at the tumor host interface. Among these activities the production of proteases could obviously influence tumor cell detachment and migration from the primary. We have evaluated the expression of plasminogen activator (PA) in different types of murine tumors with differing immunogenic capacity. In TAM from all tumors studied the expression of PA was markedly greater than that of resident peritoneal macrophages. The fact that PA was similarly enhanced in peritoneal macrophages responding to a standard stimulation (thioglycolate) and in TAM suggests that the expression of PA is part of a more general cellular inflammatory response to injury. PMID- 3047026 TI - [The autocrine regulation of growth in breast cancer]. PMID- 3047027 TI - [Myoglobin in coronary heart disease]. PMID- 3047028 TI - [Does psychotherapy work, and if so--how?]. PMID- 3047029 TI - [The newborn's erythrocyte]. PMID- 3047030 TI - [Aspects in geriatric anesthesia]. PMID- 3047031 TI - Entamoeba polecki: morphology, immunology, antigen study and clinic of the first infections in Czechoslovakia. AB - Entamoeba polecki Prowazek, 1912 was recorded for the first time in Czechoslovakia in two students from Kampuchea. Uninucleated cysts of diameter 14.2-15.7 micron with nuclei diameter 3.2-4.2 micron were found-repeatedly in stool samples taken from them. The nucleus accounted in average for 24.3% of the cyst diameter. The isolation and two following subinoculations on Dobell-Leidlaw medium were achieved, more abundant growth was recorded on medium with pig serum. Sera of both students were positive in serological tests using the E. histolytica antigen. No serious clinical symptoms were observed, both patients were cured successfully by metronidazole and ornidazole. Electroimmunotransfer blots were used to characterize E. polecki as a separate species. The antigenic structure of polyxenically grown E. polecki was compared with the antigenic structure of E. histolytica (axenically grown HK-9 strain and 3 polyxenically grown strains). Different blot patterns of both species were obtained, but common fractions of 30 40 kD probably responsible for serological cross-reactions were found. PMID- 3047032 TI - Branched chain alpha-ketoacid dehydrogenase and pyruvate dehydrogenase activity in isolated rat pancreatic islets. AB - Branched-chain alpha-ketoacid dehydrogenase and pyruvate dehydrogenase in isolated rat pancreatic islets were shown to be regulated by a phosphorylation/dephosphorylation mechanism. Broad-specificity phosphoprotein phosphatase treatment stimulated and ATP addition inhibited their activities. The kinases responsible for inactivating these complexes were shown to be sensitive to inhibition by known inhibitors, alpha-chloroisocaproate and dichloroacetate. Total activity (nmol/min/islet / 37 degrees C) of branched-chain alpha-ketoacid dehydrogenase and pyruvate dehydrogenase was 0.86 and 5.09, with a % active form (activity before phosphatase treatment divided by activity after phosphatase treatment X 100) of 36% and 94%, respectively. Incubation of intact isolated islets with alpha-chloroisocaproate affected neither insulin release nor flux through branched-chain alpha-ketoacid dehydrogenase. PMID- 3047033 TI - C-peptide during glucose loading in myxedema. PMID- 3047034 TI - Transplantation of microcarrier-attached hepatocytes into 90% partially hepatectomized rats. AB - We previously reported a method of intraperitoneal transplantation of liver cells attached to collagen-coated microcarriers, which resulted in prolonged survival and function of the transplanted cells. In the present study, we evaluated the efficacy of liver cell transplantation in providing metabolic support during acute liver insufficiency induced by 90% partial hepatectomy in rats. Ninety per cent of the liver mass (all lobes except the caudate lobe) was resected, and the rats were provided with 5% dextrose orally ad libitum upon regaining consciousness. This regimen results in severe hypoglycemia and death within 48 hr. When microcarrier-attached liver cells were transplanted into syngeneic and allogeneic recipients 3 days prior to 90% partial hepatectomy, significantly higher blood glucose levels were observed (p less than 0.01), compared to the levels in control rats which received injections of microcarriers, liver cells or medium alone. There was a marked improvement in long-term survival (40% survived longer than 28 days; p less than 0.001) in rats transplanted with microcarriers attached cells. None of the rats given injections of microcarriers, liver cells or medium alone survived beyond 5 days. When liver cells alone or attached to microcarriers were injected intraperitoneally immediately after 90% partial hepatectomy, all rats became hypoglycemic and died within 48 hr, suggesting that vascularization of the transplant is required for function of the transplanted hepatocytes. The results indicate that intraperitoneal transplantation of microcarrier-attached hepatocytes prior to 90% partial hepatectomy in rats provides acute metabolic support resulting in improved survival. PMID- 3047035 TI - Different mechanisms decrease hepatic collagen and albumin production in fasted rats. AB - Weight loss is correlated with a specific decrease in collagen synthesis in extrahepatic tissues, mainly through modulation of mRNA levels. Here, we investigated the response to weight loss in the rat liver. Male rats were either fed ad libitum or fasted for 92 hr; fasted animals lost approximately 20% of their initial body weight. Following i.p. injection of [5-3H]proline, hepatic collagen was extracted and de novo collagen production was measured. There was a decrease in the specific radioactivities of purified hepatic collagen (-75%) and albumin (-70%) relative to total hepatic protein, indicating that production of both of these proteins was specifically decreased. In fasted animals, the absolute hepatic collagen production was markedly decreased (-60%), while changes in absolute hepatic protein production were small (-15%). Using hybridization with specific DNA probes, we found that fasting causes about a 70% decrease in albumin mRNA, but the quantities of hepatic procollagen alpha 1(I) and alpha 2(I) mRNAs were unchanged. These results are consistent with regulation of albumin production during fasting by modulation of mRNA levels. The inhibition of hepatic collagen production in fasted animals, however, appears to be modulated at a posttranscriptional level or may result from increased degradation. This response differs from the pretranslational regulation of collagen synthesis in extrahepatic tissues during fasting. Furthermore, our results suggest that decreased body weight could be a potentially complicating variable in studies of collagen metabolism and fibrogenesis in the liver. PMID- 3047036 TI - Antibodies to translation products of the pre-S1 and pre-S2 regions of the envelope gene of hepatitis B virus in fulminant hepatitis B. AB - Sera from 11 patients with fulminant hepatitis B were tested for antibodies to translation products of the pre-S1 and pre-S2 regions of hepatitis B virus of IgM, IgA and IgG classes, as well as of IgA1, IgA2 and SIgA, with solid-phase enzyme immunoassays using native viral polypeptides. Antibodies to pre-S1 region product of IgM and/or IgA class were detected invariably in six patients who still had detectable hepatitis B surface antigen in serum at the time of clinical presentation. The remaining five patients who had lost HBsAg at presentation had antibodies to pre-S region products of various immunoglobulin classes in higher titers. The five patients with fulminant hepatitis without HBsAg had higher levels of IgA antibodies to pre-S region products than the seven patients with nonfulminant acute hepatitis B who had lost HBsAg: IgA antibody to pre-S1 region product (75.6 +/- 63.8 vs. 2.9 +/- 3.2, p less than 0.01) and IgA antibody to pre S2 region product (28.9 +/- 25.3 vs. 4.2 +/- 6.9, p less than 0.01). IgA antibodies to pre-S1 and pre-S2 region products were invariably polymeric in fulminant hepatitis B. These findings are compatible with the hypothesis that a heightened humoral antibody response to pre-S1 and pre-S2 region products occurs early during the course of fulminant hepatitis B, participating in severe hepatic injury and early clearance of virus characteristic of this disease. PMID- 3047038 TI - Prospects for hepatocyte transplantation. PMID- 3047039 TI - Primary hepatocyte culture: is it home away from home? PMID- 3047037 TI - Simvastatin, a competitive inhibitor of HMG-CoA reductase, lowers cholesterol saturation index of gallbladder bile. AB - We tested the possibility that simvastatin, a competitive inhibitor of HMG-CoA reductase related to mevinolin, might alter cholesterol saturation of gallbladder bile. Ten patients with Type IIa or IIb hypercholesterolemia underwent bile sampling before, and again after, treatment with 20 or 40 mg per day simvastatin for 7 to 13 weeks. Mean cholesterol saturation index of gallbladder bile fell from 1.01 to 0.77 during simvastatin treatment (p less than 0.01). This finding strongly suggests that treatment with HMG-CoA reductase inhibitors will not predispose to development of cholesterol gallstones. Indeed, it raises the possibility that such inhibitors might have a future role to play in treatment of gallstones. PMID- 3047040 TI - Hepatitis B virus infection and renal transplantation. PMID- 3047041 TI - Influence of epidermal growth factor on liver regeneration after partial hepatectomy in rats. AB - The role of epidermal growth factor on liver regeneration after partial hepatectomy in rats was investigated. After a 70% hepatectomy in rats, the concentration of epidermal growth factor in portal venous blood was unchanged compared with unoperated controls. However, small amounts of epidermal growth factor could be identified in portal venous blood after intestinal instillation of epidermal growth factor. Brunner's glands and the submandibular glands secrete epidermal growth factor. Extirpation of Brunner's glands decreased liver regeneration, whereas removal of the submandibular glands had no effect on liver regeneration. Epidermal growth factor antiserum reduced liver regeneration significantly. Oral or s.c. administration of epidermal growth factor had no effect on liver regeneration, whereas epidermal growth factor enhanced the effect of insulin and glucagon on liver regeneration. The results suggest that endogenous epidermal growth factor participates in stimulation of liver regeneration after partial hepatectomy in rats. Epidermal growth factor given together with insulin and glucagon had a synergistic effect on liver regeneration which suggests that liver regeneration in the rat is controlled by multiple regulatory peptides. PMID- 3047042 TI - The diagnostic significance of periportal hepatic necrosis and inflammation. AB - In this review the several types of cell damage and cell death which may be found in liver biopsy specimens are defined. We describe the different processes which occur at the portal/parenchymal or septal/parenchymal interface, viz. periportal spillover, periportal hepatitis, classic or lymphocytic piecemeal necrosis and biliary piecemeal necrosis. The diagnostic implications of these lesions in relation to the clinicopathological diagnosis and prognosis in various liver diseases are discussed. PMID- 3047043 TI - ras p21 oncoprotein expression in human colonic neoplasia--an immunohistochemical study with monoclonal antibody RAP-5. AB - Expression of the ras oncogene product p21 (ras p21) in benign and malignant human colonic tissues was studied using the monoclonal antibody RAP-5 and the avidin-biotin-peroxidase technique. Histologically normal colonic mucosa and hyperplastic mucosa adjacent to carcinomas (transitional mucosa) were found, in most cases, to be negative for reactivity with the antibody or showed weak staining of a few epithelial cells. Similar findings were observed in hyperplastic and juvenile polyps. Of the 145 adenomas studied, 47 (32.4%) showed detectable levels of ras p21 expression. RAP-5 immunohistochemical staining was significantly associated with the degree of epithelial dysplasia (P less than 0.01) and the size of adenoma (P less than 0.05), but not with the histological type. Fifty-four of 70 primary adenocarcinomas (77.1%) were reactive with RAP-5 and usually demonstrated a higher percentage of stained cells and more intense cytoplasmic staining than that observed in adenomas. Although metastases often displayed a similar or even higher levels of ras p21 expression compared with the primary carcinomas, in 10 cases one or more metastatic lesions showed lower levels of ras p21. These results suggest that enhanced ras p21 expression may, at times, occur in the early stages of human colon carcinogenesis but are probably not associated with metastatic tumour progression. PMID- 3047045 TI - Posttreatment use of relaxation training by cancer patients. PMID- 3047044 TI - An unusual morphological variant of follicular lymphoma. Report of two cases. AB - Most follicular lymphomas can be readily diagnosed on morphological grounds by finding closely packed pale-staining follicles partitioned by scanty dark staining interfollicular tissue. We describe two cases of an unusual 'reverse' variant of follicular lymphoma in which the nodules have dark-staining centres and pale-staining cuffs due to concentration of centroblasts at the periphery of the neoplastic follicles. Recognition of this unusual pattern of follicular lymphoma is important, to avoid confusion with progressive transformation of germinal centres or nodular lymphocyte predominance Hodgkin's disease. PMID- 3047046 TI - Psychological research in hospice care: toward specificity of therapeutic mechanisms. PMID- 3047047 TI - Rural hospitals double private-pay rates to cover Medicare shortfall. PMID- 3047048 TI - Battle cry for hospitals: 'Elect to protect'. PMID- 3047049 TI - HHS outlines 'safe harbor' business practices. PMID- 3047050 TI - Migration and genetic change. Raymond Pearl lecture 1987. PMID- 3047051 TI - Acute rejection in human liver grafts: a comparative histologic study of cases maintained on azathioprine and prednisone versus cyclosporine A and low-dose steroids. AB - The morphology of acute rejection (AR) in biopsies of liver allografts obtained in the first 2 weeks after transplantation was analyzed. Material from patients maintained on azathioprine and prednisone (AZA; Groningen, The Netherlands) was compared with that of patients receiving cyclosporine A and prednisone (with or without azathioprine) in low doses (CSA; Minneapolis). Strict selection criteria were applied to exclude circulatory and biliary complications and viral infection in this early observation period after transplantation. Follow-up biopsies ranged from 3 weeks to 1 year after transplantation. Time zero biopsies and/or pretransplant biopsies served as baseline histology, Our data revealed an identical morphologic picture during AR early after transplantation in both patient groups, except for a more marked degree of venous endothelialitis and hepatocyte ballooning in the Minnesota material. The follow-up biopsies suggested a spontaneous resolution of these early rejection episodes without antirejection treatment in six of the ten AZA patients. No differences in the long-term survival rate between the CSA- and AZA-treated patients were observed. PMID- 3047052 TI - The histopathology of the hemolytic uremic syndrome associated with verocytotoxin producing Escherichia coli infections. AB - Verocytotoxin-producing Escherichia coli (VTEC) infection was present in three cases of hemolytic uremic syndrome (HUS), two fatal and one non-fatal, in which detailed histopathologic investigations were conducted. Two patients had a prodrome of bloody diarrhea, one of whom required a hemicolectomy for severe bleeding. The renal histopathology was characterized primarily by glomerular thrombotic microangiopathy (TMA) with greater than 95% of glomeruli showing changes of capillary wall thickening, endothelial cell swelling, and narrowing or thrombosis of the capillary lumen. Preglomerular arterioles were frequently thrombosed, and abnormalities of the medium-sized vessels, including endothelial cell damage and thrombosis, were also commonly observed. Gastrointestinal involvement was prominent in all three cases. The colon was most severely involved, with marked mucosal and submucosal edema and hemorrhage, in the absence of significant inflammation or widespread ulceration. Microvascular angiopathy was present in all cases, with changes ranging from endothelial cell damage to overt thrombosis. Similar pathology was seen throughout the small bowel, including the presence of TMA. In one patient, typical morphologic changes of pseudomembranous enterocolitis were found in the absence of infection with Clostridium difficile. The nature of vascular involvement in the kidneys and intestinal tract supports the hypothesis that HUS is mediated by systemic toxemia, and that endothelial cells are the primary target cells for the action of verocytotoxin. PMID- 3047053 TI - ras p21 expression in the progression of breast cancer. PMID- 3047054 TI - Monoclonal antibody RAP-5 and ras p21 expression. PMID- 3047055 TI - Mastering the library maze. PMID- 3047056 TI - Imprint publishers directory: resource books for nursing students. PMID- 3047057 TI - [Results of a multicenter study on the use of beta-interferon in the treatment of mucocutaneous lesions caused by herpes simplex virus. A Multicenter Study Group]. PMID- 3047058 TI - [Hyperkeratosis, deafness and keratitis (KID syndrome). Study of 3 cases with dermatophytosis and review of the literature]. PMID- 3047059 TI - Inhibition of granulocyte function by steroids is not limited to corticoids. Studies with sex steroids. AB - Noting that corticosteroid doses required for protection in shock models exceeded those required to saturate glucocorticoid receptors in mammalian cells, we postulated a nonspecific physicochemical effect of steroids upon the cell membrane, and therefore tested three noncorticoid steroids for their effects on granulocyte function. All three (conjugated equine estrogen, a synthetic progestogen, and a synthetic androgen) behaved in manner analogous to corticoids at similar concentrations, inhibiting granulocyte aggregation, chemotaxis, and chemiluminescence, as well as binding to the granulocytes of the synthetic oligopeptide agonist f-Met-Leu-Phe. Estrogen was further shown to reduce granulocyte membrane fluidity, assessed by electron paramagnetic resonance. We propose that the unique effects of extremely high-dose corticosteroids are not mediated via the glucocorticoid receptor, but result rather from physicochemical effects of the drugs upon the membranes of effector cells. PMID- 3047060 TI - Inhibition of penetration of cultured cells by Eimeria bovis sporozoites by monoclonal immunoglobulin G antibodies against the parasite surface protein P20. AB - Five monoclonal antibodies (MAbs) were partially characterized and tested for their ability to inhibit penetration of Madin-Darby bovine kidney (MDBK) cells by sporozoites of Eimeria bovis. By indirect fluorescent-antibody assays, all MAbs reacted with acetone-fixed sporozoites, but only two MAbs, EbS9 (immunoglobulin G1) and EbS11 (immunoglobulin G2a), localized specifically on the plasmalemma of live sporozoites. Two of the five MAbs also reacted with acetone-fixed first generation merozoites of E. bovis; however, none of the MAbs reacted with live merozoites. Treatment of live sporozoites with EbS9 or EbS11 resulted in 79 and 73% decreases, respectively, in sporozoite penetration of MDBK cells. No significant differences in cell penetration occurred in MDBK cells inoculated with sporozoites that had been treated with the other three MAbs. Both EbS9 and EbS11 reacted in Western blots (immunoblots) of sporozoites with the same 20,000 relative-molecular-weight protein. The antigens against which these neutralizing MAbs react might be useful in immunizing against bovine coccidiosis. PMID- 3047061 TI - In vitro expression of a 22-kilodalton Yersinia pestis polypeptide immunologically related to the 25-kilodalton plasmid-encoded protein of the three pathogenic Yersinia species. AB - Antibodies raised against the 25-kilodalton (p25) plasmid-encoded polypeptide of Yersinia enterocolitica recognized the homologous protein in the three Yersinia species grown in vitro. This polypeptide was recovered from whole cells as well as from the fluid supernatant of bacteria grown at 37 degrees C in a Ca2+ deficient medium. Furthermore, a 22-kilodalton (p22) plasmid-encoded polypeptide immunologically related to p25 was found only in Y. pestis during early growth. After 30 h of culture, the Y. pestis p25 and p22 were completely degraded, whereas the intensity of the Y. enterocolitica p25 was decreased, but the protein was still detectable in the fluid supernatant. This proteolytic activity was independent of the presence of the virulence plasmid. Some disulfide bonds are probably involved in the quaternary structure of the p25 of the three pathogenic species and of the Y. pestis p22. PMID- 3047063 TI - Immunoreactivity of a surface wall fraction produced by spherules of Coccidioides immitis. AB - The membranous spherule outer wall (SOW) isolated from liquid cultures of Coccidioides immitis has been shown to elicit reactivity with human anti Coccidioides antibody by immunofluorescence and the immunodiffusion-tube precipitin assay. The serologically reactive components were extracted from SOW with the nonionic detergent N-octyl-beta-D-glucopyranoside (OG). The OG-soluble fraction of SOW was shown to be reactive with immunoglobulin G in 25 serum samples from coccidioidomycosis patients by an enzyme-linked immunosorbent assay. The isolated SOW and OG-soluble fraction of SOW were also demonstrated to be capable of eliciting lymphocyte blastogenesis. The antigenic and protein compositions of the OG-soluble fraction were examined by two-dimensional immunoelectrophoresis and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), respectively. Two antigens which were extracted from SOW were identified as antigens 2 and CS on the basis of the coccidioidin anticoccidioidin reference system. The latter was isolated earlier and shown to correspond to a molecular mass (Mr) of 19 X 10(3) by SDS-PAGE under reducing conditions. This same electrophoresis band was shown to be reactive with sera from coccidioidomycosis patients by immunoblot analysis. One other SDS-PAGE component of the OG-soluble fraction of SOW with an Mr of 66 X 10(3) was shown to be reactive with sera from patients by immunoblot analysis. The SOW of C. immitis represents an important reservoir of immunoreactive wall components which has not previously been reported. PMID- 3047062 TI - Effect of carrier selection on immunogenicity of protein conjugate vaccines against Plasmodium falciparum circumsporozoites. AB - Conjugate vaccines against the sporozoite stage of Plasmodium falciparum were synthesized by covalently coupling the recombinant protein R32 [with the one letter amino acid code of MDP-[(NANP)15NVDP]2LR] to tetanus toxoid, cholera toxin, choleragenoid, and Pseudomonas aeruginosa toxin A. Conjugates were produced by using adipic acid dihydrazide as a spacer molecule and carbodiimide as a coupling agent. The molar ratio of R32 to carrier protein ranged from 2.5:1 to 8.4:1. These conjugates were found to be stable, nontoxic, and nonpyrogenic. When adsorbed onto Al(OH)3, all conjugates were capable of inducing anti-R32 antibody. Conjugates made with either cholera toxin or Pseudomonas aeruginosa toxin A were significantly more immunogenic than those constructed with tetanus toxoid or choleragenoid. However, the magnitude of the immune response to the R32 moiety was not governed by the antibody response to the carrier protein. PMID- 3047065 TI - Immunolabeling of lipopolysaccharide liberated from antibiotic-treated Escherichia coli. AB - Increased anti-core glycolipid antibody binding was visualized by immunoelectron microscopy after incubation of Escherichia coli K1:O7 cells with a bacteriolytic antibiotic and compared with binding in control cells. The findings suggest that the core glycolipid regions of the lipopolysaccharides of some gram-negative bacilli can be effectively sequestered until liberated by antibiotic-induced cell lysis. PMID- 3047064 TI - Quantitative relationship between capsular content and killing of K1-encapsulated Escherichia coli. AB - Since there are conflicting reports in the literature on a possible relationship between the K1 capsular polysaccharide (CP) content of Escherichia coli and its susceptibility to killing, we reexamined this issue in a strain that had a smooth lipopolysaccharide (LPS) phenotype (E. coli O18:K1:H7 Bort) and in a strain with a deep rough LPS phenotype (E412, spontaneously agglutinable: K1:H-). When cell associated K1 capsular content was greater than 90 micrograms of K1 polysaccharide per 10(10) CFU, neither strain was lysed by 20% normal human serum. In contrast, at equivalent but lower levels of K1 CP content, E412 but not strain Bort was lysed by normal human serum. Thus, LPS phenotype is an additional surface determinant that affects bacterial susceptibility to killing. Organisms obtained from very early log phase, when cell-associated K1 CP is greatest, were significantly more virulent for mice than were bacteria harvested in stationary phase, when cell-associated K1 polysaccharide is lowest. We conclude that (i) there is a threshold level of K1 CP needed to confer protection from lysis by serum, and this is usually exceeded under standard growth conditions; (ii) at a given level of K1 CP the LPS phenotype is an important determinant of bacterial killing; and (iii) the loss of capsule at low pH may be an additional mechanism by which hosts defend against invasive infection by K1-encapsulated E. coli. PMID- 3047066 TI - Antibiotic susceptibilities of Listeria: in vitro studies. AB - Although Listeria is a rather susceptible bacterium, most antibiotics exert a bacteriostatic effect on Listeria monocytogenes. Except for fosfomycin, antibiotic susceptibilities are similar among the species of the genus Listeria. In vitro, bactericidal effect is often achieved by the use of antibiotic combinations. The most commonly used combinations are ampicillin with aminoglycosides. Up until now, there has been no trend towards reduced susceptibility of Listeria to antibiotics. PMID- 3047067 TI - Listeria monocytogenes infections--therapeutic possibilities and problems. AB - Listeriosis in humans is a rare disease, which, however, is known to be epidemic and endemic. The prognosis has remained unsatisfactory up to today, the fatality being at least 10% and often considerably higher depending on the clinical features of the disease and the patient's age. Three population groups are at risk: pregnant women, fetuses and newborn infants. Furthermore, immunosuppression in older patients due to disease, therapy, or age also plays a role. The incidence of Listeria infections in patients over 45 is clearly increasing. Due to the nature of the pathogen (in vivo bactericidal concentrations of antibiotics are often not attainable; intracellular growth) a high dosage of ampicillin is recommended. Although the present therapeutic possibilities are not satisfactory, a combination of ampicillin and an aminoglycoside appears to be the best therapy at present. Other combinations such as rifampicin and beta-lactam antibiotics have exhibited in vitro antagonism. The preferred therapy, ampicillin, can only be recommended with reservations because it is not optimally effective. PMID- 3047068 TI - Isolation of Listeria: a review of procedures and future prospects. AB - Recent documented foodborne outbreaks of listeriosis have underscored the need for improved isolation and identification procedures for Listeria. This review considers the development of selective enrichment media, various approaches to improving efficiency of isolation, and efforts to shorten enrichment periods. The application of simplified rapid nucleic acid hybridization techniques, in combination with improved cultural methods is the most promising of these approaches. Such methodological improvements should facilitate gathering data on important questions concerning the epidemiology of Listeria, and the natural history of listeriosis. PMID- 3047070 TI - Parasitological societies of the world. PMID- 3047069 TI - In celebration of the 65th birthday of Robert Maxwell and the 40th anniversary of Pergamon Press. PMID- 3047071 TI - Centenary biographical note. Robert-Philippe Dollfus 1887-1976. PMID- 3047072 TI - Monoclonal antibodies specific for Onchocerca volvulus as determined by immunofluorescence. PMID- 3047073 TI - Neonatal ophthalmia in the developing world. Epidemiology, etiology, management and control. AB - In the 19th century, the incidence of neonatal conjunctivitis varied between 1 and 14% in Europe, and the disease was a main cause of blindness at that time. Since then the epidemiology of ophthalmia neonatorum (ON) has changed and Chlamydia trachomatis is more frequent than Neisseria gonorrhoeae. Both are still very common causes of ON in the developing world. ON can not be differentiated clinically as to the etiology, but Intracellular Gram Negative Diplococci (IGND) on a Gram stain of an eye smear has an excellent validity and further differentiation can be made using microbiological cultures. All cases of presumed gonococcal conjunctivitis must be treated with effective systemic antibiotics. Systemic treatment with penicillin can still be used in areas where the percentage of beta-lactamase producing strains of gonococci is very low. For other areas a single dose of ceftriaxone intramuscular combined with saline eye washes is the treatment of choice. Chlamydial ON necessitates also systemic treatment with erythromycin. Parents of infants with gonococcal or chlamydia ON also need to be examined and treated. Prevention of gonococcal and chlamydial disease can be done following 3 strategies: antenatal diagnosis and treatment of maternal infections or disinfection of the infants eyes at birth or adequate treatment of infants and parents as soon as a ON has been diagnosed. Crede's eye prophylaxis with silver nitrate has become a controversial issue, because of concern about the occurrence of chemical conjunctivitis and its ineffectiveness against infections with C. trachomatis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3047075 TI - Zacharias Dische 2/18/1895-1/17/1988. PMID- 3047074 TI - The extracellular matrix composition of the monkey optic nerve head. AB - The presence and distribution of laminin, heparan sulfate proteoglycan and collagen types I, III and IV were immunohistologically determined in cynomolgus monkey optic nerve heads using the avidin-biotin-peroxidase complex technique. Collagen types I and III were detected within the collagenous plates of the scleral lamina cribrosa, in the septa and pia mater of the postlaminar optic nerve and in blood vessel walls in all regions of the optic nerve head. Collagen type IV, laminin and heparan sulfate proteoglycans were all localized to the margins of the collagenous laminar plates of the scleral lamina cribrosa and along the margins of the optic nerve septa and the pia mater. All three components also appeared beneath the blood vessel endothelium throughout the optic nerve head. Within the lamina cribrosa, collagen types I and III occupy the core of the scleral laminar plates and may provide structural support for optic nerve bundles exiting the eye. The distribution of collagen type IV, laminin and heparan sulfate proteoglycan corresponds to basement membranes from two sources: vascular endothelial cells and glial cells lining the axonal bundles. Abnormalities of these substances may influence optic nerve function and susceptibility to elevated intraocular pressure by altering their mechanical support functions within the nerve head, by interfering with axonal nutrition, or both. PMID- 3047076 TI - Stimulation of endothelial cell prostacyclin release by retina-derived factors. AB - An angiogenic extract of bovine retina as well as two purified angiogenic growth factors, acidic and basic fibroblast growth factor, stimulate vascular endothelial cell prostacyclin (PGI2) release in vitro as measured by radioimmunoassay of its stable metabolite 6-keto PGF1 alpha (6kPGF). After incubating fetal bovine aortic endothelial cells with 10% retinal extract (RE) for 24 hr, 6.5 ng of 6-kPGF/10(5) cells were released compared to 0.8 ng of 6kPGF/10(5) cells for unstimulated endothelium. Similar qualitative results were obtained using human retina-derived microvessel endothelium. PGI2 release in response to RE depended on endothelial cell density with subconfluent cultures releasing six-fold more 6kPGF compared to confluent monolayers. Thin layer radiochromatography of endothelial cell conditioned media demonstrated enhanced release of 6-kPGF, PGE2 and arachidonic acid after RE addition. Cycloheximide and actinomycin D inhibited PGI2 release but hydroxyurea had no effect. Most of the PGI2-stimulating activity of RE was adsorbed to heparin-Sepharose and eluted by 2 M NaCl. Purified acidic (10 ng/ml) and basic (1 ng/ml) fibroblast growth factors caused seven-fold and four-fold stimulation of endothelial PGI2 release, respectively. An initial event in the regression of new blood vessels following the removal of an angiogenic stimulus is the formation of intraluminal platelet aggregates. PGI2 is a potent inhibitor of platelet aggregation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3047077 TI - Isoproterenol treatment causes cytoskeletal reorganization in chicken lens fiber cells. AB - Cytoskeletal organization in organ cultured embryonic chicken lens was affected by isoproterenol treatment. Immunofluorescent localization of a 49 kD fiber cell specific cytoskeletal protein showed a cytoplasmic and membrane localization in control lenses. Following isoproterenol treatment, fluorescence was predominantly membrane-associated. Changes in 49 kD distribution as determined by immunofluorescence occurred within minutes of isoproterenol application and were completely blocked by propranolol pretreatment. These results suggest that beta adrenergic stimulation influences the apparent localization of a fiber cell specific cytoskeletal protein. PMID- 3047078 TI - Tissue culture study of adult human retina neurons. AB - Explant cultures were established from adult human retina tissues obtained from 20 individuals (age range 19-79), 7-45 hr postmortem, and maintained for the period of up to 4 months. Forty percent of these cultures (eight out of 20 cases) produced healthy and viable cultures as judged by phase contrast microscopy and by electron microscopy. Phase contrast microscopic examination of living cultures showed that the early outgrowths of the explants consisted of flat fibroblasts migrating out from the edge of the explants. For the next 2-4 weeks, a large population of small spherical or ovoid cells possessing thin processes was found in the areas of the outgrowth. Electron microscopic examination of cultures revealed the survival of photoreceptors, neurons and synapses with well preserved ultrastructures. This communication is the first to describe the successful culture of adult human CNS neurons in general and adult human retina neurons in particular. This culture system is ideally suited to investigate the effects of infective or toxic agents suspected of causing retinal pathology in human eye diseases. PMID- 3047079 TI - Murine models of Sjogren's syndrome. Immunohistologic analysis of different strains. AB - Lacrimal gland inflammation develops in several strains of autoimmune mice, including MRL/Mp-lpr/lpr (MRL/lpr), MRL/Mp-+/+ (MRL/+), and NZBxNZW F1 hybrids (NZB/W). These mice all develop an autoimmune disease characterized by glomerulonephritis and autoantibody formation, but each strain has unique clinical features and immunologic abnormalities. Previous studies have suggested that the intrinsic immunologic defect in MRL/lpr mice may be at the level of T cells, while in NZB/W mice it appears to be B cell-mediated. Immunohistologic analysis of the lacrimal gland lesions was performed on all three strains. Although T cells predominated (MRL/lpr 85%, MLR/+ 78%, and NZB/W 57%), differences in the immunohistologic profiles did exist. NZB/W mice had a significantly higher percentage of B cells (33% vs. 10% for MRL/lpr and 13% for MRL/+) and a correspondingly lower percentage of T cells. MRL/lpr mice differed from MRL/+ mice in that they exhibited a significantly higher percentage of helper T cells (63% vs. 49%) and a lower percentage of suppressor/cytotoxic T cells (14% vs. 30%). Class II antigen expression could be detected on the mononuclear cells at inflammatory sites within the lacrimal glands of all three strains, suggesting T cell activation and an active autoimmune immunologic event occurring in the lacrimal gland. PMID- 3047080 TI - Effects of dilating the canine pulmonary annulus with two balloons. Doppler ultrasound and postmortem findings. AB - Simultaneous inflation of two balloons may be necessary for balloon dilation valvuloplasty in patients with a large annulus. We examined the cardiovascular effects of dilating the pulmonary annulus with two balloons in 18 normal dogs using pulsed Doppler ultrasound and gross and microscopic examination. When the ratio of the cross-sectional area (CSA) during dilation to the CSA of the pulmonary annulus was between 1.04 and 1.28, there was valvar regurgitation in only one dog, in which catheter manipulation was complicated by heartworms, and damage was confined to intimal changes. (The dog was killed 24 hours after dilation.) With ratios between 1.67 and 1.76 (equal to the CSA of a single balloon with a diameter 33% greater than the annulus), there was trivial or mild tricuspid or pulmonary regurgitation, and anatomic changes were more prominent but still superficial. Two animals killed after nine days had resolution of valvar regurgitation and healing damage. With CSA ratios greater than 2.00 or with balloon rupture, myocardial damage and laceration of the pulmonary arteries resulted. Simultaneous inflation of two balloons within the right ventricular outflow tract with CSA ratios of up to 1.76 results in minimal cardiovascular trauma. PMID- 3047081 TI - X-rays at the bar, 1896-1910. PMID- 3047082 TI - Esophageal pressure monitoring: a practical adjuvant to hemodynamic monitoring with positive end-expiratory pressure. AB - With positive end-expiratory pressure (PEEP)-induced reduction in cardiac output, measurement of ventricular filling pressure assists in proper therapeutic decision-making. Because PEEP may increase pleural and juxtacardiac pressure, central venous pressure (CVP) and left atrial pressure (LAP) measurements during PEEP may not simply reflect ventricular filling, but rather reflect the sum of intracardiac and extracardiac forces. Monitoring devices placed within the central circulation use saline solution-filled lumens and transducer systems for pressure monitoring. Therefore, any device designed to estimate the extracardiac influence of PEEP on intraluminal monitoring devices would be expected to reflect such changes best when the device is also filled with saline solution. In the present study, esophageal pressure (Pes) was measured with a saline solution filled balloon-equipped nasogastric tube to estimate the extracardiac influence of PEEP on CVP and LAP. Pes, CVP, LAP, and cardiac index (CI) were measured in 17 patients subjected to 0, 5, 10, 15, 20 cm H2O PEEP. Comparing 0 with 20 cm H2O PEEP, CVP (7 +/- 1.0 mm Hg to 13.4 +/- 1.3 mm Hg), LAP (6.3 +/- 1.1 mm Hg to 11.7 +/- 1.4 mm Hg), and Pes (6.1 +/- 1.4 mm Hg to 12.1 +/- 1.5 mm Hg) all increased significantly as CI fell (2.72 +/- 0.14 L/min/m2 to 2.20 +/- 0.15 L/min/m2).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3047083 TI - Cardiac transplantation: a comprehensive nursing perspective. Part 1. AB - Orthotopic cardiac transplantation has become the treatment of choice for selected patients with end-stage heart disease. Proliferation of cardiac transplantation centers and relaxed selection criteria have resulted in increasing populations of transplant candidates and recipients. As these numbers continue to grow, so too do the numbers of nursing personnel who must respond to the intricacies and demands of cardiac transplantation. Nursing care begins with physiologic and psychologic support of a patient with terminal cardiac disease and continues throughout the transplantation evaluation, waiting, surgical, postoperative, and outpatient periods. In addition to routine postsurgical care, the nurse involved in cardiac transplantation needs to be familiar with the function of a denervated heart, as well as with the effects of unbalanced immunosuppression and immunocompetence. The surge of organ transplantation in the last several years has brought about administrative concerns regarding the financial impact of increasing numbers of transplantations, as well as an inadequate supply of necessary donor organs. This critical shortage of suitable donors demands that nurses and other health care professionals become active in enlarging the donor organ pool by identifying potential organ donors, maintaining optimal hemodynamics and oxygenation so as to ensure organ viability, and providing emotional support of the grieving family. PMID- 3047084 TI - Rewarming and cardiac surgery: a review. AB - Patients undergoing cardiac surgery are mildly hypothermic by the completion of the surgical procedure. They need to return to a normothermic state if enzymatic functions are to proceed in their normal manner. The body can produce heat by elevating metabolic rate or by activating the shivering mechanism. Metabolic rate peaks shortly after separation of the patient from cardiopulmonary bypass, and therefore contributes to heat production. Because of the effects of neuromuscular blockage administered both during and after surgery; these patients may be unable to generate heat by shivering, and shivering is usually undesirable. This eliminates the major heat production mechanism available to the body. Therefore, heat must be transferred down its gradient by means of convection and conduction. External and internal methods accomplish these goals. External methods, which minimize additional heat loss, include the use of warming lights, elevation of room temperature, and the use of blankets. Internal methods, which transfer heat by convection, may be used to help actively reverse hypothermia. Such techniques include warmed inhalation gases and intravenous fluids, warmed nasogastric lavage fluid, and warmed peritoneal dialysis fluid for patients with end-stage renal failure with severe electrolyte disorders after surgery. PMID- 3047085 TI - Pulmonary valvuloplasty as an alternative to surgery in the pediatric patient: implications for nursing. AB - Recent innovations in the use of balloon catheters to dilate stenosis have made it possible to successfully perform balloon valvuloplasty (BV) on stenotic intracardiac valves. BV is fast becoming the treatment of choice for isolated valvular pulmonary stenosis (IVPS) in pediatric patients. IVPS and other obstructive lesions involving the right ventricle and pulmonary arteries occur in 25% to 30% of all persons with congenital heart disease. Current data suggest that use of BV for IVPS provides both short- and long-term hemodynamic relief and eliminates the need for open heart surgery. BV is also being used for other forms of congenital heart disease. Therefore, it is important for nurses in the pediatric setting to be aware of this technique and its implications. We review the anatomy, physiology, and clinical features of IVPS, the valvuloplasty procedure, and the nursing plan of care relevant to it. PMID- 3047086 TI - The impact of AIDS in the pediatric and adolescent populations. PMID- 3047087 TI - Taking the lid off eating disorders. PMID- 3047088 TI - Does adjuvant radiotherapy have a role in the postmastectomy management of patients with operable breast cancer--revisited. AB - About 2 decades ago, "routine" adjunctive postmastectomy radiotherapy, especially for axillary node-positive patients, was the norm and uncriticized standard against which adequate treatment was measured in most centers. With the advent of cyclic, aggressive, multi-agent chemotherapy and anti-hormones used as adjuvants, especially within the last decade, there has been tremendous reduction in patients referred to the radiation oncologist for consideration of adjunctive postmastectomy radiotherapy. This presentation will attempt to define a role for radiotherapy in at least selected subsets of patients who undergo modified radical mastectomy, based upon published series in the literature. Breast cancer is a protean disease and deserves a multidisciplinary approach to evaluation and treatment. "Routine" adjunctive postmastectomy radiotherapy for all patients with operable breast cancer obviously is not indicated, but there appear to be groups of patients who benefit from radiotherapy, both from the standpoint of disease free survival and improved quality of life, and . . . in very narrow subsets, absolute survival. In some of these subsets the benefit clinically may be greater than that resulting from chemotherapy or anti-hormone therapy, although, because of sample size, falling short of statistical verification. In response to the posed question, while this remains a controversial issue, there appears to be a role for selective adjunctive postmastectomy radiotherapy in specific subsets of patients, and physicians administering adjunctive breast cancer therapy with sweeping applications of chemotherapy or anti-hormones alone do not appear to be offering their patients optimal therapy. PMID- 3047089 TI - The Radiation Therapy Oncology Group--1987. PMID- 3047090 TI - Low-dose-rate total lymphoid irradiation: a new method of rapid immunosuppression. AB - Total Lymphoid Irradiation (TLI) has been successful in inducing immunosuppression in experimental and clinical applications. However, both the experimental and clinical utility of TLI are hampered by the prolonged treatment courses required (23 days in rats and 30-60 days in humans). Low-dose-rate TLI has the potential of reducing overall treatment time while achieving comparable immunosuppression. This study examines the immunosuppressive activity and treatment toxicity of conventional-dose-rate (23 days) vs low-dose-rate (2-7 days) TLI. Seven groups of Lewis rats were given TLI with 60Co. One group was treated at conventional-dose-rates (80-110 cGy/min) and received 3400 cGy in 17 fractions over 23 days. Six groups were treated at low-dose-rate (7 cGy/min) and received total doses of 800, 1200, 1800, 2400, 3000, and 3400 cGy over 2-7 days. Rats treated at conventional-dose-rates over 23 days and at low-dose-rate over 2 7 days tolerated radiation with minimal toxicity. The level of immunosuppression was tested using allogeneic (Brown-Norway) skin graft survival. Control animals retained allogeneic skin grafts for a mean of 14 days (range 8-21 days). Conventional-dose-rate treated animals (3400 cGy in 23 days) kept their grafts 60 days (range 50-66 days) (p less than .001). Low-dose-rate treated rats (800 to 3400 cGy total dose over 2-7 days) also had prolongation of allogeneic graft survival times following TLI with a dose-response curve established. The graft survival time for the 3400 cGy low-dose-rate group (66 days, range 52-78 days) was not significantly different from the 3400 cGy conventional-dose-rate group (p less than 0.10). When the total dose given was equivalent, low-dose-rate TLI demonstrated an advantage of reduced overall treatment time compared to conventional-dose-rate TLI (7 days vs. 23 days) with no increase in toxicity. This was accomplished without compromise of the immunosuppressant activity of TLI as demonstrated by comparable allogeneic skin graft survival times between the two 3400 cGy treatment groups. This clinical advantage would prove to be beneficial where immediate suppression of the immune system is desirable. PMID- 3047092 TI - Assessing the role of adjuvant radiation therapy in the treatment of breast cancer. PMID- 3047091 TI - Single dose or fractionated total body irradiation and autologous marrow transplantation in dogs: effects of exposure rate, fraction size, and fractionation interval on acute and delayed toxicity. AB - Dogs were given single dose or fractionated total body irradiation (TBI) and autologous marrow grafts to prevent death from myelosuppression. Acute and delayed non-marrow toxicities were compared. Fifty-six dogs were given single dose TBI at 2.1 (n = 13), 5 (n = 12), 10 (n = 15), or 20 (n = 16) cGy/min. Acute radiation toxicity and mortality was related to the exposure rate; radiation doses resulting in 50% mortality at 7 days (LD 50/7) at 2.1, 5, 10 and 20 cGy/min were 1,692, 1,499, 1,261, and 1,056 cGy respectively. Fifty-three dogs were given fractionated TBI, 200 cGy three times a day with 6-hour intervals at 2.1 (n = 13), 5 (n = 9), 10 (n = 13), or 20 (n = 18) cGy/min. The LD 50/7 at the four exposure rates were 1,628, 1,470, 1,184, and 1,320 respectively. Thus, for exposure rates of 2.1, 5, and 10 cGy/min, the tolerated doses were comparable for single dose and fractionated TBI. At 20 cGy/min dose fractionation appeared to offer some advantage, although this fractionation effect in part may have been due to random variation with small numbers of dog treated. Following recovery from the immediate TBI-related toxicity, eight dogs given single dose and four dogs given fractionated TBI died, generally from infections, 8-30 days following transplantation. There was a striking difference in regards to long-term survival dependent upon the TBI regimen. Among dogs given greater than or equal to 1,000 cGy of TBI and alive 30 days after transplant only 1 of 18 given single dose TBI became a long-term survivor compared to 19 of 22 given fractionated TBI. Causes of death included pancreatic fibrosis, malnutrition, hepatic failure, and a generalized wasting syndrome. All 5 dogs given a single dose of 800 cGy (at 20 cGy/min) became long-term survivors. Fourteen dogs were given increments of 150 cGy at 7 cGy/min every 3 hours for total doses of 1,500-2,400 cGy. The LD 50/7 was approximately 1,900 cGy. All 6 dogs alive at 30 days became healthy long-term survivors. Four dogs were given increments of 600 cGy at 2.1 cGy/min every 48 hours for a total dose of 1,800 cGy. All 4 dogs became long-term survivors. In conclusion, exposure rate and total dose are the most important parameters for acute toxicity associated with TBI. The effect of dose fractionation is minimal at low exposure rates and appears to be dependent also upon increment size and fractionation interval.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3047093 TI - Use of enzyme immunoassay for the rapid diagnosis of Chlamydia trachomatis endocervical infection in female adolescents. AB - Enzyme immunoassay (EIA) has been proposed as an alternative to tissue culture for the detection of Chlamydia trachomatis in cervical specimens. The diagnostic efficacy of EIA was compared to tissue culture in 113 teenaged females attending an adolescent reproductive health program. Infection was diagnosed by tissue culture in 16% of subjects. Compared with tissue culture, EIA demonstrated a sensitivity of 100%, specificity of 88%, positive predictive value of 62%, and a negative predictive value of 100%. These data indicate that EIA is an acceptable alternative to tissue culture when screening for C. trachomatis endocervical infection in adolescent females. PMID- 3047094 TI - Fellowships in Adolescent Medicine--1988. PMID- 3047095 TI - Bovine practice in the Texas Panhandle. PMID- 3047096 TI - Analysis of the dental morphology of Plio-Pleistocene hominids. IV. Mandibular postcanine root morphology. AB - The subocclusal morphology of 168 permanent mandibular premolars (N = 77) and molars (N = 91) of Plio-Pleistocene hominids has been investigated. The taxonomic allocation of the teeth, which represent at least 46 individuals, was based on nondental evidence. Specimens were allocated to one of two major taxonomic categories, (EAFROB or EAFHOM), East African Homo erectus (EAFHER), or their taxonomic affinity was regarded as 'unknown' (N = 17). Information about the root system was derived from radiography and direct observation. Morphometric data were in the form of nine linear and two angular measurements based on eighteen reference points. Root form was also assessed using a scheme which recognised four classes of root morphology. Data were compared using both univariate and multivariate techniques, including Principal Component and Canonical Variate analysis. Posterior probabilities derived from the latter were used (in a two taxon design model) to assess the affinities of the 'unknown' specimens. The variation in hominid mandibular premolar root form was interpreted as two morphoclines, based on the presumed primitive condition of the P3 (with mesiobuccal and distal roots, 2R: MB and D) and P4 (with mesial and distal root, 2R: M and D) root systems. One trend apparently leads towards root reduction (i.e. P3 = 1 R; P4 = 1 R), and the other to root elaboration (i.e. P3 and P4 = 2R: M and D). The extreme form of the latter is the 'molarisation' of the premolar roots seen in EAFROB. Despite major differences in root form there was relatively little taxonomic variation in root metrics, except for a more robust distal root system in EAFROB. Molar root form showed little interspecific variation except for M2 in which the roots in EAFROB were larger and more robust, with differences in root height being greater for the distal than for the mesial roots. Root form and metrics enable four of the 'unknown' specimens (KMN-ER 819, 1482, 1483 and 1801) to be tentatively allocated to EAFHOM, and a single specimen, KMN-ER 3731, to EAFROB. Published assessments of the root morphology of the 'robust' australopithecines from Swartkrans suggest that the premolar root form of Australopithecus (Paranthropus) robustus is not obviously intermediate between the presumed ancestral condition, and the 'molarised' mandibular premolar root systems of Australopithecus (Paranthropus) boisei. PMID- 3047097 TI - Immunofluorescent analysis of somatotroph distribution in the adenohypophysis of developing lit/lit mice. AB - Regional and sexual patterns in the distribution and density of somatotroph cells exposed to anti-growth hormone serum were analysed by means of immunofluorescence histochemistry in adenohypophyses of normal C57BL mice and abnormal (lit/lit) mutant mice, which exhibit postnatal growth deficits. In adult (3-4 months) lit/lit mice, the regional distribution of somatotrophs both in males and females was normal; however, there was a sparsity of somatotrophs, relative to the normal condition, in the lateral wings of the pars distalis, and sexual differences in the concentration of immunoreactive cells were not as prominent as in the normal mice. In the midline region of the pars distalis a cranioventral zone virtually devoid of somatotrophs occurred in lit/lit as well as in normal mice, especially in the females, though it was not as well defined in lit/lit because of the overall sparsity of somatotrophs. In normal immature mice at 8 and 14 days after birth, the lateral wings did not show the striking sexual differences in density of somatotroph distribution as they did in the normal adults, and at 8 days they were more sparsely populated with somatotrophs than at 14 days. In 14 day lit/lit mice, the lateral wings were less densely populated with somatotrophs than their normal counterparts, but at 8 days these differences were not detectable. In both normal and abnormal 8 day mice, the medial and midline regions of the pars distalis contained less intensely immunoreactive somatotrophs than did the lateral wings.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3047099 TI - Evaluation of amino acid analysis as reference method to quantitate highly purified proteins. AB - The applicability of amino acid analysis for accurate quantitation of reference standard preparations of proteins has been evaluated. This approach is very useful since, in addition to absolute quantitative information, it also provides a measure of composition, partial identity, and purity in a single experiment. Comparisons with Kjeldahl nitrogen assay and/or UV measurements shows that amino acid analysis is reliable for the quantitation of small-to-medium size proteins in the molecular weight range of 6-22 kDa. For larger proteins such as immunoglobulins (150 kDa), amino acid analysis may "underestimate" the total protein concentration. These results also show the effect of recovery of individual residues on protein quantitation. As expected, the recovery of more than one stable residue could be used to calculate total protein content of samples, which is in good agreement with the results obtained by Kjeldahl nitrogen assay. However, the protein concentrations calculated from the total mass of the recovered residues appear to give relatively low estimates in almost all cases. Thus, it is concluded that amino acid analysis is an appropriate reference method only when stable residues are employed for quantitation, particularly for highly purified proteins of rDNA origin. PMID- 3047098 TI - Review of the decontamination of aflatoxins by ammoniation: current status and regulation. AB - Ammoniation of corn, peanuts, cottonseed, and meals to alter the toxic and carcinogenic effects of aflatoxin contamination has been the subject of intense research effort by scientists in various government agencies and universities, both in the United States and abroad. Results of these studies have been well documented over the last 20 years. Engineers have devised workable systems of treatment of whole seeds, kernels, or meals; chemists have identified and characterized products formed from the reaction of aflatoxin B1 with ammonia with and without a meal matrix; biochemists have studied the biological effects of these compounds in model systems; and nutritionists have studied animal responses to rations containing ammoniated or non-ammoniated components. This review describes these studies. Results demonstrate overwhelming support for the efficacy and safety of ammoniation as a practical solution to aflatoxin detoxification in animal feeds. PMID- 3047101 TI - Update on alcohol and drug disorder treatment. AB - The numbers of patients in all forms of psychoactive substance abuse disorder treatments have dramatically risen in the past decade, and it is estimated that $1.8 billion are spent on alcohol and drug inpatient and outpatient treatment. Recent developments in therapeutic modalities and the differential therapeutics of alcohol and psychoactive drug abuse treatment are presented. Choice of treatment setting, combination of treatment modalities, and the special needs of subpopulations are discussed in terms of both review of the literature and practical decisions made by clinicians. Problems in diagnosis and treatment and the dual-diagnosis patient, patients with multiple substance abuse, women, minorities, and the homeless are touched upon. The heterogeneous nature of patients with psychoactive substance abuse problems requires flexibility and sophistication in treatment design, with attention to biopsychosocial variables. The treatment implications of recent developments in diagnosis such as DSM-III-R, research on AIDS in relation to substance abuse, and studies of familial alcoholism and fetal alcoholism are highlighted. A concise update of detoxification, pharmacotherapy, behavioral treatment, psychodynamically informed approaches, family therapy, and group therapy focuses on clinical decisions. Although demonstrating overall efficacy of alcohol treatment, few studies demonstrate differential responses for subpopulations or specific treatment modalities. PMID- 3047102 TI - The treatment of eating disorders. AB - Anorexia nervosa and bulimia nervosa are common disorders among adolescent and young adult females, estimated to occur in up to 1% to 3% of these populations. A variety of medical and psychological approaches have been used in their treatment, including hospitalization; nutritional rehabilitation; medications; and psychodynamic, behavioral, cognitive-behavioral, family, and group psychotherapies. The author discusses controlled and uncontrolled studies of the different therapeutic approaches that have been used in these disorders and reviews what is known about their prognosis. Important treatment issues include the nature of the disorder, age, degree of malnutrition, motivation, capacity to engage in treatment, personality disturbance, affective state, family situation, and available treatment resources. Based on research data and clinical lore, prudent suggestions for treatment planning are presented. PMID- 3047100 TI - Profile of mood states changes during and after 5 weeks of nightly triazolam administration. AB - This double-blind, placebo-controlled study investigated rebound anxiety during and after 5 weeks of nightly use of triazolam 0.5 mg, a short half-life, rapidly absorbed benzodiazepine hypnotic. The study subjects were chronic insomniacs with moderate levels of psychopathology and prior use of hypnotics. Anxiety was assessed with the Profile of Mood States (POMS), instead of the anxiety scales more typically used in psychopharmacology: the Hamilton Rating Scale for Anxiety and a visual analogue scale. The POMS test was administered twice a day during the study. The results indicated that triazolam was not associated with increased anxiety the morning or the evening after previous-night drug administration. The results are discussed in view of the methodological issues in assessing anxiety in prior studies. PMID- 3047103 TI - Psychodynamic treatment of depressed adolescents. AB - Depression may be conceptualized as the response to the loss of meaning or satisfaction sufficient to affect the individual's optimal view of the self. At each stage of the life cycle, the failure to achieve developmental tasks threatens the concept of the self, producing a phase-specific vulnerability to depression. Adolescence presents particular stresses by forcing the youngster to relinquish the relative familiarity and security of a childhood psychosocial role and create a sense of self independent of family, without childhood denial mechanisms, and of value to a new peer culture. Most individuals experience a sense of loss, confusion, apprehension, and dysphoria during this difficult period of transition. Many who seek psychotherapy require only a secure holding environment that will support their self-esteem as they create new avenues of worth and satisfaction. A few, however, are so hampered by psychosocial limitations that they cannot master this developmental passage without more extensive therapeutic assistance. PMID- 3047104 TI - Populations at high alcoholism risk: recent findings. AB - This article reviews recent studies of men at high risk for the future development of alcoholism. The most promising differences between young men with and without a family history of this disorder include a decreased intensity of reaction to ethanol for sons of alcoholics compared with control subjects, a decreased amplitude of the P300 wave of the event-related potential, and a different pattern of background cortical electroencephalograms (alpha waves) for subjects at high risk for future alcoholism. The implications of these and other findings and their possible future impact on the clinical practice are discussed. PMID- 3047106 TI - The epidemiology of teen suicide: an examination of risk factors. AB - The information presented here is on the incidence of suicide in the United States and in other countries where reporting procedures are reliable. The problem of underreporting is discussed and put into perspective. Secular trends in suicide incidence are presented, and it is shown how these have varied a great deal for different age groups. Age, ethnic, and cultural differences in incidence are demonstrated, and explanatory theories for these differences are put forward. The phenomenon of cluster suicide is described, and possible explanations for this are discussed. The limitations of death certificate data and the advantages (and limitations) of the psychological autopsy method are presented. Results from previous psychological autopsy studies on epidemiologically sound samples are summarized. Results (preliminary) from a current psychological autopsy study on consecutive adolescent suicides in the New York Metropolitan area are presented. Family history data and diagnostic profiles of completed suicides are emphasized. PMID- 3047107 TI - Suicide prevention in depressed women. AB - This paper focuses on risk factors and clinical states associated with suicide in depressed women and the necessity for thorough clinical evaluation of family and personal histories of suicidal, impulsive, and violent behaviors. Treatment issues addressed include the importance and content of detailed communication to patients and their families, medication noncompliance, the risks associated with the recovery and other high-risk periods, and problems associated with the adjunctive use of alcohol and other drugs. PMID- 3047105 TI - Treatment of borderline conditions in adolescents. AB - The history of the borderline concept is reviewed and its diagnostic criteria described. Current views regarding etiology and pathogenesis are presented with a summary of treatment. This includes highlights of individual and family psychotherapy, aspects of hospitalization, and a presentation of the current status of pharmacologic interventions. PMID- 3047108 TI - What destroys our restraints against suicide? AB - There seems to exist an inherent barrier to the achievement of any extreme biological state, be it massive intravascular coagulation, intense euphoria, or the inalterable decision to kill oneself. Moreover, defenses against suicide may in themselves be pathological, producing other types of maladaption. Indeed, certain lethality indicators possess a threshold, below which they act as a barrier against suicide. Wrist-cutting, self-treatment with drugs and alcohol, and psychosis itself all can play a defensive role in the organism's struggle for survival. PMID- 3047109 TI - Alcoholism: clinical implications of recent research. AB - The alcoholism field has traditionally been marked by strong divisions between research and clinical practice. The most common modes of clinical practice, rooted in a strong self-help tradition, have not seemed to gain much from biological or behavioral research. Indeed, the most famous (and controversial) behavioral research study of the 1970s appeared to undermine the efforts of clinicians to encourage abstinence as the only reasonable treatment goal for alcoholics. Another study conducted in England in the 1970s appeared to show that simple one-time advice worked as well as more intensive, traditional treatment interventions in persuading alcoholic individuals to modify their behavior. In the 1980s, studies on DSM-III-defined co-morbid psychopathology have served to redefine the heterogeneity of alcoholic patients in treatment, while describing the impact of co-morbidity on clinical course and treatment response. Measures of severity of alcohol dependence in clinical populations led directly to the development of the DSM-III-R criteria for this disorder. Indeed, these measures of severity may have predictive validity relative to treatment response. Finally, newer pharmacological strategies have been suggested, based upon animal and preliminary human studies, which may reduce risk of relapse in some patients. Reviews of the current state of clinically significant research in the alcohol field, with implications for clinicians and clinical investigators are presented. PMID- 3047110 TI - The history of Eduard Pernkopf's Topographische Anatomie des Menschen. PMID- 3047111 TI - Histone-like proteins and bacterial chromosome structure. PMID- 3047112 TI - Mutations that simultaneously alter both sugar and cation specificity in the melibiose carrier of Escherichia coli. AB - The isolation and deduced amino acid sequence of 70 melibiose carrier mutants with impaired methyl-beta-D-galactopyranoside (TMG) and cation recognition properties is described. The Km for TMG transport ranged from 1 to greater than 100 mM. Amino acid substitutions occurred at 23 unique sites within the protein. These sites were clustered into four distinct regions: Asp-15 through Ile-18 (cluster I), Tyr-116 through Pro-122 (cluster II), Val-342 through Ile-348 (cluster III), and Ala-364 through Gly-374. Only two sites fell outside of these clusters: Ile-61 and Ala-236. In the native conformation, some or all of these clusters may interact to form the substrate recognition site. Impairment of TMG recognition was accompanied by decreased Li+ inhibition of melibiose transport in all but one mutant. That changes in sugar recognition properties should so frequently accompany changes in cation recognition properties suggests an interaction between the two substrates. A model for such interaction is proposed. PMID- 3047113 TI - Insulin increases the synthetic rate and messenger RNA level of lipoprotein lipase in isolated rat adipocytes. AB - Lipoprotein lipase (LPL) is the enzyme responsible for hydrolysis of circulating triglyceride-rich lipoproteins and is important for storage of adipocyte lipid. To study the regulation of LPL synthetic rate in adipose tissue, primary cultures of isolated rat adipocytes were pulse-labeled with [35S]methionine, and LPL was immunoprecipitated with an LPL-specific antibody. A pulse-chase experiment identified the cellular and secreted forms of LPL as a 55-57-kDa protein. In the presence of heparin, there was a large increase in secretion of newly synthesized LPL from the cells, although heparin did not stimulate cellular LPL synthetic rate. When cells were exposed to insulin for 2 h, pulse-labeling revealed that insulin stimulated a maximal dose-related increase in LPL synthetic rate of 300% of control. This increase in LPL synthetic rate was observed after an exposure to insulin for as little as 60 min and was accompanied by only a 10-25% increase in total protein synthesis. In addition, insulin had no effect on the turnover of intracellular LPL. Using a cDNA probe for LPL, insulin induced a 2-fold increase in the LPL mRNA. Thus, insulin stimulated an increase in specific LPL mRNA in isolated rat adipocytes. This increase in LPL mRNA then leads to an increase in the synthetic rate of the LPL protein. PMID- 3047114 TI - In vitro processing of pro-subtilisin produced in Escherichia coli. AB - In a previous paper (Ikemura, H., Takagi, H., and Inouye, M. (1987) J. Biol. Chem. 262, 7859-7864), we demonstrated that the pro-sequence consisting of 77 amino acid residues at the amino terminus of subtilisin is essential for the production of active subtilisin. When the aggregates of pro-subtilisin produced in Escherichia coli were solubilized in 6 M guanidine hydrochloride and dialyzed against 200 mM sodium phosphate buffer (pH 7.1 or 6.2), pro-subtilisin was efficiently processed to active subtilisin. When more than 14 residues were removed from the amino terminus of the pro-sequence, active subtilisin was no longer produced as in the in vivo experiments. Similarly, active subtilisin would not renature under the same conditions once solubilized in guanidine hydrochloride. When the aspartic acid residue at the active site (Asp32) was altered to asparagine, processing of mutant pro-subtilisin was not observed even in the presence of wild-type pro-subtilisin. Inhibitors such as phenylmethanesulfonyl fluoride or Streptomyces subtilisin inhibitor did not block the processing of wild-type pro-subtilisin. These facts indicate that processing or pro-subtilisin is carried out by an intramolecular, self-processing mechanism. When the sample was dialyzed against 20 mM sodium phosphate (pH 6.2), no active subtilisin was found, suggesting that the highly charged nature of the pro sequence plays an important role in the process of refolding of denatured pro subtilisin. PMID- 3047115 TI - Pathway of phospholipase C activation initiated with platelet-derived growth factor is different from that initiated with vasopressin and bombesin. AB - The mode of phospholipase C activation initiated with platelet-derived growth factor (PDGF) has been studied in comparison with that initiated with vasopressin and bombesin in a rat fibroblast line, WFB. Stimulation of WFB cells by PDGF, vasopressin, and bombesin elicites rapid hydrolysis of polyphosphoinositides and an increase in cytoplasmic free Ca2+ concentration ([Ca2+]i). On stimulation by PDGF, there was a lag period of about 10 s before an increase in [Ca2+]i. No measurable lag period was observed in the [Ca2+]i response induced by vasopressin or bombesin. Pretreatment of WFB cells with phorbol 12-myristate 13-acetate profoundly inhibited inositol phosphate formation evoked by vasopressin and bombesin, but enhanced to some extent inositol phosphate formation stimulated by PDGF. In membranes prepared from WFB cells, GTP markedly augmented inositol polyphosphate formation induced by vasopressin and bombesin. It was not successful in showing the PDGF-stimulated formation of inositol phosphates in the membrane preparation. The effects of GTP, guanosine 5'-O-(2-thiodiphosphate) (GDP beta S), and guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S) on polyphosphoinositide hydrolysis stimulated by growth factors were studied in WFB cells made permeable to nucleotides by treatment with either saponin or Pseudomonas aeruginosa cytotoxin. PDGF, vasopressin, and bombesin elicited inositol phosphate production in the permeabilized WFB cells in the absence of added GTP. GDP beta S, a competitive inhibitor of GTP-binding proteins (G proteins), markedly reduced the bombesin- and vasopressin-stimulated production of inositol phosphates. However, the PDGF-stimulated production of inositol phosphates was not affected by the addition of GDP beta S. GTP gamma S, an agonist of G-proteins, largely enhanced the vasopressin- and bombesin-stimulated hydrolysis of inositol lipids when added at 10-100 microM. In the presence of GTP gamma S, the PDGF-stimulated hydrolysis of inositol lipids was not enhanced, but was reduced: 100 microM GTP gamma S reduced the stimulated hydrolysis to about a half of the control level. Only GTP gamma S, and no other nucleoside triphosphates, was found to have these effects. Activation of G-proteins in WFB cells by fluoroaluminate resulted in the inhibition of inositol phosphate production elicited with not only PDGF, but also with vasopressin and bombesin. These results indicate that a G-protein couples vasopressin and bombesin receptors to the activation of phospholipase C. Moreover, these results suggest that coupling of the PDGF receptor to phospholipase C is not mediated through a G protein.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3047116 TI - Glucose permease of Escherichia coli. Purification of the IIGlc subunit and functional characterization of its oligomeric forms. AB - The membrane subunit (IIGlc) of the glucose permease has been purified from overproducing Escherichia coli. About 2 mg of pure protein was obtained from 10 g (wet weight) of cells. IIGlc of E. coli and Salmonella typhimurium are functionally indistinguishable. A small difference was revealed, however, by a monoclonal antibody which neutralizes glucose phosphorylation activity of IIGlc from S. typhimurium, but does not cross-react with IIGlc of E. coli. A dimeric form of purified IIGlc can be detected by chemical cross-linking and by zonal sedimentation at 4 degrees C. Upon mild oxidation a disulfide bond is formed between the subunits of the dimer. Oxidized IIGlc is more stable than the reduced form but is inactive because it cannot be phosphorylated by the cytoplasmic subunit (IIIGlc) of the glucose permease. Cys-421 could be identified as the oxidation-sensitive residue, using a novel assay to detect IIIGlc-dependent phosphorylation of nitrocellulose-bound IIGlc that has been purified by gel electrophoresis. No dimeric form of phosphorylated IIGlc could be detected. Because phosphorylated IIGlc is a catalytic intermediate it is concluded that catalytically active IIGlc is a monomer and that the dimeric form is an artefact observed only with purified resting IIGlc. That IIGlc is active as a monomer is further supported by the observation that monomeric IIGlc catalyzes phosphoryl exchange between glucose and glucose 6-phosphate at equilibrium and that an excess of inactive IIGlc with a serine replacing Cys-421 does not interfere with the activity of wild-type IIGlc as would be expected if interaction between the subunits in a dimer were essential for activity. PMID- 3047117 TI - The role of an active site histidine in the catalytic mechanism of aspartate transcarbamoylase. AB - Although the allosteric enzyme aspartate transcarbamoylase from Escherichia coli has been the focus of numerous physical and enzymological studies for over 2 decades, the catalytic mechanism is still poorly understood. There has been much speculation regarding the role of a conserved histidine residue at position 134 which recent crystallographic studies have implicated in the catalytic mechanism as a general acid or as a general base. We have used a combination of site directed mutagenesis, 13C-isotope incorporation, and high field NMR to probe the role of His134 in the catalytic mechanism of aspartate transcarbamoylase. By comparing the wild-type catalytic trimer with that from a partially active mutant in which His134 is replaced by alanine, we have assigned the 13C resonance for His134 in the wild-type enzyme. This residue is shown to have a pK less than 6, indicating that the imidazole ring is unprotonated at pH values optimal for enzymatic activity (pH 8.0). This result eliminates the possibility of His134 participating as a general acid in the carbamoyl transfer mechanism. Since the crystallographic studies indicate that His134 is close enough to hydrogen-bond to the carbonyl of the liganded bisubstrate analog N-(phosphonacetyl)-L-aspartate, the imidazole ring would be oriented as to make it unlikely that the N1 lone pair of electrons could participate in general base catalysis. Moreover, if His134 is implicated in base catalysis, we would have expected a much greater loss of activity upon its replacement by alanine. Perhaps the role of His134 is merely to help position the carbonyl group of carbamoyl phosphate for nucleophilic attack by the alpha-amino group of aspartate. PMID- 3047118 TI - Hormonal regulation of insulin receptor gene expression. Hydrocortisone and insulin act by different mechanisms. AB - Incubation of cultured human (IM-9) lymphocytes with glucocorticoids increases the number of insulin receptors on the cell surface. Recently it has been shown that this results from a 3-fold increase in the rate of proreceptor synthesis. In the case of homologous insulin receptor down-regulation, a moderate rise in proreceptor biosynthesis has also been demonstrated. To further delineate the mechanisms of insulin receptor regulation, we have measured insulin receptor mRNA levels in hydrocortisone-treated and insulin-treated IM-9 lymphocytes. An increase in insulin receptor mRNA could be detected after 2 h of incubation with hydrocortisone and a plateau was reached by 4-6 h. The response was dose dependent, being detectable with 50 nM hydrocortisone and reaching a maximal 3.7 fold increase at 200 nM. Actinomycin D completely suppressed the effect, whereas cycloheximide inhibited the effect by no more than 50%. These findings were extended by performing in vitro nuclear transcription assays which revealed that the 3-4-fold increase in insulin receptor mRNA could be attributed to increases in transcription. In contrast, homologous down-regulation was not associated with any change in total insulin receptor mRNA levels. The present study demonstrates that hydrocortisone, but not insulin, stimulates insulin receptor biosynthesis by increasing the rate of transcription and that de novo protein synthesis is probably required for a maximal effect. PMID- 3047119 TI - Human cytotoxic lymphocyte tryptase. Its purification from granules and the characterization of inhibitor and substrate specificity. AB - A trypsin-like enzyme (tryptase) has been purified to homogeneity from the granules of a human cytolytic lymphocyte (CTL) line, Q31, by a three-step procedure. By including 0.3% (v/v) Triton X-100 and 1 mg/ml heparin in purification buffers, near total yields of tryptase activity were obtained during the purification. The enzyme, referred to as Q31 tryptase, migrated in polyacrylamide gels with sodium dodecyl sulfate at a position corresponding to 28 kDa with and to 45 kDa without 2-mercaptoethanol. It had an amino-terminal sequence identical to a previously reported human CTL tryptase at 20 of 22 positions identified. It hydrolyzed N alpha-carbobenzyloxy-L-lysyl-thiobenzyl ester (BLT), and this BLT esterase activity was most efficient at slightly alkaline pH and was relatively more active near neutral pH than mouse CTL tryptase. Human alpha 1-protease inhibitor, human antithrombin III, phenylmethanesulfonyl fluoride, and p-aminobenzamidine inhibited the Q31 tryptase. The inhibition by human antithrombin III was rapid enough to be of physiological significance. A survey of oligopeptide p-nitroanilides found that the best substrate for human Q31 tryptase is H-D-(epsilon-carbobenzyloxy)Lys-L Pro-L-Arg-p-nitroanilide. The Q31 tryptase appears to have broad specificity for amino acid residues at P2 and P3, i.e. at 2 and 3 residues amino-terminal to the scissile bond. PMID- 3047120 TI - Internal deletions in the gene for an Escherichia coli outer membrane protein define an area possibly important for recognition of the outer membrane by this polypeptide. AB - A series of overlapping deletions has been constructed in the ompA gene which encodes the 325-residue Escherichia coli outer membrane protein OmpA. Immunoelectron microscopy showed that the OmpA fragments were either located in the periplasmic space or were associated with the outer membrane. Apparently an area between residues 154 and 180 is required for this association; all proteins missing this area were found to be periplasmic. The nature of this association remained unknown; no membrane-protected tryptic fragments could be identified for any of these polypeptides. Hybrid genes were constructed encoding parts of the periplasmic maltose binding protein and an area of the ompA gene coding for residues 154-274. The corresponding proteins were not localized to the outer membrane but remained attached to the outer face of the plasma membrane, possibly because the normal mechanism of release from this membrane was impaired. In the OmpA protein the conspicuous sequence Ala180-Pro-Ala-Pro-Ala-Pro-Ala-Pro187 exists. Frameshift mutants were constructed to eliminate this sequence. There was no effect on the incorporation of the mutant proteins into the outer membrane. Thus, this "hinge" region is not involved in sorting. A proposal suggesting the existence of a sorting signal common to several outer membrane proteins (Benson, S. A., Bremer, E., and Silhavy, T. J. (1984) Proc. Natl. Acad. Sci. U. S. A. 81, 3830-3834) was subsequently rejected (Bosch, D., Leunissen, J., Verbakel, J., de Jong, M., van Erp, H., and Tommassen, J. (1986) J. Mol. Biol. 189, 449-455; Freudl, R., Schwarz, H., Klose, M., Movva, N. R., and Henning, U. (1985) EMBO J. 4, 3593-3598). Although it is not known whether or not the outer membrane association observed represents a step in the normal sorting mechanism, it is concluded that it remains an open question whether or not a sorting signal, as proposed originally, exists in outer membrane proteins. PMID- 3047121 TI - The influence of amino substitutions within the mature part of an Escherichia coli outer membrane protein (OmpA) on assembly of the polypeptide into its membrane. AB - The membrane part of the 325-residue outer membrane protein OmpA of Escherichia coli encompasses residues 1-177. This part is thought to cross the membrane eight times in antiparallel beta-strands, forming four loops of an amphipathic beta barrel. With the aim of gaining some insight into the mechanism of sorting, i.e. the way the protein recognizes and assembles into its membrane, a set of point mutants in the ompA gene has been generated. Selection for toxicity of ompA expression following mutagenesis with sodium bisulfite yielded genes with multiple base pair substitutions, the majority of which resulted in amino acid substitutions in the membrane moiety of the protein. None of the altered proteins was blocked in membrane incorporation. A proline residue exists at or near each of the presumed turns at the inner side of the outer membrane. Using oligonucleotide-directed mutagenesis, each of them was replaced by a leucine residue which is thought to be a turn blocking residue. None of these proteins had lost the ability to be incorporated into the membrane. Apparently, leucine residues are tolerated at turns in this protein. To interfere with the formation of antiparallel beta-strands, four double mutants were prepared: ompA-ON3 (Ala11- --Pro, Leu13----Pro), -ON4 (Ala11----Asp, Leu13----Pro), -ON5 (Gly160----Val, Leu162----Arg), and -ON6 (Leu164----Pro, Val166----Asp). The former three proteins and even quadruple mutants consisting of a combination of ompA-ON2 or ON4 with -ON5 were not defective in membrane assembly. In contrast, the OmpA-ON6 protein was translocated across the plasma membrane but could not be incorporated into the outer membrane. It is concluded that at least one rather small area of the polypeptide is of crucial importance for the assembly of OmpA into the outer membrane. PMID- 3047122 TI - Characterization of the human transferrin receptor produced in a baculovirus expression system. AB - Recombinant human transferrin receptor has been produced in a baculovirus expression system. Magnetic particles coated with an anti-transferrin receptor monoclonal antibody were used to immunoselect virus-infected Sf9 insect cells expressing the human transferrin receptor on their cell surface. Recombinant virus containing the human transferrin receptor cDNA was then plaque-purified from these cells. Biosynthetic labeling studies of infected cells showed that the human transferrin receptor is one of the major proteins made 2-3 days postinfection. The recombinant receptor made in insect cells is glycosylated and is also posttranslationally modified by the addition of a fatty acid moiety. However, studies with tunicamycin and endoglycosidases H and F showed that the oligosaccharides displayed on the recombinant receptor differ from those found on the naturally occurring receptor in human cells. As a consequence, the human receptor produced in the baculovirus system has an Mr of 82,000 and is smaller in size than the authentic receptor. About 30% of human transferrin receptors made in insect cells do not form intermolecular disulfide bonds, but are recognized by the anti-transferrin receptor antibody, B3/25, and bind specifically to a human transferrin-Sepharose column. Binding studies using 125I-labeled human transferrin showed that insect cells infected with the recombinant virus expressed an average of 5.8 +/- 0.9 X 10(5) transferrin receptors (Kd = 63 +/- 9 nM) on their cell surface. Thus, the human transferrin receptor produced in insect cells is biologically active and appears suitable for structural and functional studies. PMID- 3047123 TI - A latent proteinase in mouse kidney membranes. Characterization and relationship to meprin. AB - Inbred mice can be phenotypically divided into two groups: those that contain high levels of a kidney metallo-endopeptidase activity (meprin-a) and those with low meprin-a activity. In studies to investigate the molecular basis for the heterogeneity in the expression of this proteinase activity, we found a latent metallo-proteinase activity associated with kidney membranes of C3H/HeJ mice, a low activity strain. The latent proteinase was activated by treatment of kidney brush border membranes with trypsin and was purified from solubilized C3H kidney membranes. Purified preparations of the C3H latent proteinase (referred to as meprin-b) contained three major proteins of subunit molecular weights 90,000, 140,000, and 160,000. In the absence of reducing agents, four 90,000-Da subunits are covalently linked by S-S bridges. The two higher molecular mass proteins are not covalently linked to each other or to the 90,000-Da subunits. However, cross linking and affinity chromatography studies indicated that the proteins in the meprin-b preparation were tightly associated. By contrast, purified meprin-a contains only 85,000-Da subunit proteins linked by S-S bridges to form a tetramer. Endoglycosidase F treatment decreased the mass of the 90,000-Da meprin b subunit and the 85,000-Da meprin-a subunit to polypeptides of 65,000-70,000 Da. The 90,000- and 85,000-Da subunits are immunologically similar, in that polyclonal antibodies prepared against one of the subunits cross-react with the other. The substrate specificities and inhibitor profiles of purified preparations of meprin-a and meprin-b are also similar. These data are consistent with the proposition that meprin-b is a polymorphic form of meprin-a that is incompletely processed in vivo. PMID- 3047124 TI - Studies with antipeptide antibody suggest the presence of at least two types of glucose transporter in rat brain and adipocyte. AB - Three antipeptide antibodies were prepared by immunizing rabbits with synthesized short peptides corresponding to residues 215-226, 466-479, and 478-492 predicted from the cDNA of both the human hepatoma HepG2 and rat brain glucose transporters. All three antibodies were found to precipitate quantitatively the [3H]cytochalasin B photoaffinity-labeled human erythrocyte glucose transporter. Each antibody also recognized the rat brain protein of Mr 45,000 on immunoblots, and a similar molecular weight protein was labeled with [3H]cytochalasin B in a D glucose-inhibitable manner, suggesting that this protein is glucose transporter. However, only up to 30% of the labeled rat brain glucose transporters were precipitated, even by repeated rounds of immunoprecipitation. In addition, these antibodies were observed to be unable to immunoprecipitate significantly the [3H]cytochalasin B-labeled rat adipocyte glucose transporter. Further, one dimensional peptide maps of [3H]cytochalasin B-labeled human erythrocyte and adipocyte glucose transporters generated distinct tryptic fragments. Although Mr 45,000 protein in rat adipocyte low density microsomes was detected on immunoblots and its amount was decreased in insulin-treated cells, the rat adipocyte low density microsomes were much less reactive on immunoblots than the rat brain membranes in spite of the fact that the rat adipocyte low density microsomes contained more [3H]cytochalasin B-labeled glucose transporters. In addition, the ratio of cytochalasin B-labeled glucose transporter per unit HepG2 type glucose transporter mRNA was more than 10-fold higher in rat adipocyte than in rat brain. These results indicate that virtually all the human erythrocyte glucose transporters are of the HepG2 type, whereas this type of glucose transporter constitutes only approximately 30 and 3% of all the glucose transporters present in rat brain and rat adipocyte, respectively; and the rest, of similar molecular weight, is expressed by a different gene. PMID- 3047125 TI - Stimulation of prolactin gene expression by insulin. AB - GH3 cells are a rat pituitary-derived cell line in which the expression of the growth hormone and prolactin genes is controlled by a variety of hormones including thyroid hormone. Since these cells contain insulin receptors, we explored whether these cells can be used to analyze the mechanisms involved in the regulation of gene expression by insulin. When GH3 cells were incubated with serum-free media, insulin stimulated prolactin production rates about 3-fold after 72 h and 10-fold after 96 h of incubation. Insulin stimulated prolactin mRNA levels and gene transcription rates to the same extent as the prolactin production rates. In contrast, insulin did not stimulate the rate of growth hormone production or growth hormone mRNA levels. As previously reported, thyroid hormone stimulated growth hormone production and mRNA levels. In these cells, thyroid hormone also stimulated prolactin synthesis and prolactin mRNA levels. Prolactin production and mRNA levels in cells incubated with both insulin and thyroid hormone showed a synergistic response which was about 5- to 10-fold greater than with either hormone alone. In contrast, growth hormone production rates in cells cultured with insulin and thyroid hormone was only slightly greater than cells incubated with only thyroid hormone. Half-maximal stimulation of prolactin production in cells incubated with insulin or with both insulin and thyroid hormone occurred between 2 and 5 nM insulin, suggesting that the response is mediated by the insulin receptor. These results indicate that GH3 cells should be useful in analyzing the mechanisms involved in regulation of gene expression by insulin. PMID- 3047126 TI - Differential binding of the chicken ovalbumin upstream promoter (COUP) transcription factor to two different promoters. AB - The COUP (chicken ovalbumin upstream promoter) transcription factor binds to the upstream promoters of both the chicken ovalbumin and the rat insulin II genes, even though the two binding sites have no obvious sequence similarity (Hwung, Y. P., Crowe, D. T., Wang, L.-H., Tsai, S. Y., and Tsai, M.-J. (1988) Mol. Cell. Biol. 8, 2070-2077). Comparison of the contact points and important nucleotides in the two binding sites suggests that the COUP transcription factor binds to them in different ways. The purine contacts in the two promoters are limited to one face of the DNA helix; however, studies on phosphate contacts suggest that the COUP transcription factor wraps around the ovalbumin promoter, while it binds to only one face of the DNA helix in the insulin promoter. In addition, the binding factor makes a more extended contact on the insulin promoter as compared to the ovalbumin promoter. The potential significance of this novel finding is discussed. PMID- 3047127 TI - Effects of amino acid substitutions in the F helix of bacteriorhodopsin. Low temperature ultraviolet/visible difference spectroscopy. AB - Site-specific mutagenesis in combination with low temperature UV/visible difference spectroscopy has been used to investigate the role of individual amino acids in the structure and function of bacteriorhodopsin (bR). We examined the effects of eight single amino acid substitutions, all in the putative F helix, on the absorption of bR as well as formation of the K and M intermediates. Both the absorbance spectra and the photocycle difference spectra of Escherichia coli expressed bR as well as the mutants S183A, P186G, and E194Q all closely resembled the corresponding purple membrane spectra. In contrast the Pro-186----Leu substitution resulted in the loss of the normal photocycle and a large blue shift in the bR state lambda max. Thus, Pro-186 appears to play a critical role in maintaining the normal protein-chromophore interactions, although the pyrrolidine ring is not essential since proline could be replaced by glycine at this position. The mutants W182F, W189F, and S193A did not appear to be directly involved in the bathochromic shift of bR since they all had lambda max's close to that of purple membrane and produced intermediates similar to K and M. However, alterations in the UV and visible difference spectra as well as the appearance of some irreversibility in the photoreactions indicate that these mutants have altered protein-chromophore interactions during the photocycle. Unlike the other mutants examined, Y185F exhibited a red-shifted form of bR and K raising the possibility that Tyr-185 is directly involved in color regulation. In addition, UV difference peaks previously associated with a tyrosine deprotonation were absent in Y185F indicating that Tyr-185 undergoes protonation changes during the photocycle in agreement with recent Fourier transform infrared difference measurements (Braiman, M.S., Mogi, T., Stern, L. J., Hackett, N., Chao, B. H., Khorana, H.G., and Rothschild, K. J. (1988) Proteins: Structure, Function, and Genetics 3, 219-229). Our results suggest that Trp-182, Tyr-185, Pro-186, Trp 189, and Ser-193, all of which are within a 100 degrees segment of the F helix, are part of a retinal-binding pocket. PMID- 3047128 TI - Regulation of insulin release by factors that also modify glutamate dehydrogenase. AB - Leucine and monomethyl succinate initiate insulin release, and glutamine potentiates leucine-induced insulin release. Alanine enhances and malate inhibits leucine plus glutamine-induced insulin release. The insulinotropic effect of leucine is at least in part secondary to its ability to activate glutamate oxidation by glutamate dehydrogenase (Sener, A., Malaisse-Lagae, F., and Malaisse, W. J. (1981) Proc. Natl. Acad. Sci. U. S. A. 78, 5460-5464). The effect of these other amino acids or Krebs cycle intermediates on insulin release also correlates with their effects on glutamate dehydrogenase and their ability to regulate inhibition of this enzyme by alpha-ketoglutarate. For example, glutamine enhances insulin release and islet glutamate dehydrogenase activity only in the presence of leucine. This could be because leucine, especially in the presence of alpha-ketoglutarate, increases the Km of glutamate and converts alpha ketoglutarate from a noncompetitive to a competitive inhibitor of glutamate. Thus, in the presence of leucine, this enzyme is more responsive to high levels of glutamate and less responsive to inhibition by alpha-ketoglutarate. Malate could decrease and alanine could increase insulin release because malate increases the generation of alpha-ketoglutarate in islet mitochondria via the combined malate dehydrogenase-aspartate aminotransferase reaction, and alanine could decrease the level of alpha-ketoglutarate via the alanine transaminase reaction. Monomethyl succinate alone is as stimulatory of insulin release as leucine alone, and glutamine enhances the action of both. Succinyl coenzyme A, leucine, and GTP are all bound in the same region on glutamate dehydrogenase, where GTP is a potent inhibitor and succinyl coenzyme A and leucine are comparable activators. Thus, the insulinotropic properties of monomethyl succinate could result from it increasing the level of succinyl coenzyme A and decreasing the level of GTP via the succinate thiokinase reaction. PMID- 3047129 TI - Alpha-glucan phosphorylase from Escherichia coli. Cloning of the gene, and purification and characterization of the protein. AB - By using a synthetic oligonucleotide probe identical to a part of the gene for the Escherichia coli major outer membrane lipoprotein, we have cloned a gene from E. coli chromosomal DNA. However, the cloned gene was not one of the lipoprotein genes. The amino acid sequence deduced from its nucleotide sequence shows extensive similarities instead to alpha-glucan phosphorylase (EC 2.4.1.1). The gene, glgP, is located immediately downstream from glgA, the gene for glycogen synthase. The glgP gene was inserted into pUC9 vector and expressed in the presence of the lac inducer. The gene product was purified to apparent homogeneity as shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In all chromatographies, the protein was eluted accompanied by a low phosphorylase activity. The final preparation showed phosphorolytic activity to various alpha-glucans, although the specific activity was extremely low compared to other alpha-glucan phosphorylases under the standard assay conditions. Its enzymatic activity, however, increased almost linearly as the concentration of glucan increased, reaching a value comparable with those of other phosphorylases. The amino acid sequence deduced was compared with those of alpha-glucan phosphorylases from other sources. PMID- 3047130 TI - Nutrient secretagogues induce bimodal early changes in cytoplasmic calcium of insulin-releasing ob/ob mouse beta-cells. AB - The cytoplasmic calcium concentration (Ca2+i) was measured in suspensions of fura 2 loaded mouse pancreatic beta-cells by continuously recording the 340/380 nm fluorescence excitation ratio. When the glucose concentration was raised from 3 to 20 mM, there was an initial lowering of Ca2+i followed by a sustained increase. Whereas the reduction in Ca2+i was related to the extracellular glucose concentration in a hyperbolic manner, the increasing component exhibited a sigmoidal dose-response relationship. Both effects became maximal at 15-20 mM of the sugar. Qualitatively similar bimodal Ca2+i responses were obtained with 30 mM mannose, 2 mM alpha-ketoisocaproic acid, 10 mM leucine, and 10 mM metabolism stimulating leucine analogue beta-2-aminobicyclo-(2,2,1)-heptane-2-carboxylic acid. Fructose (30 mM) had virtually no effect on Ca2+i in the presence of extracellular Ca2+, and 10 mM arginine induced only a rise. The results indicate that nutrient secretagogues stimulate both the entry of Ca2+ into the beta-cells and its elimination from the cytoplasm by processes like organelle sequestration and outward transport. Consequently, the Ca2+i level determining insulin secretion results from the balance between two opposing actions. PMID- 3047131 TI - Glenoid bone-grafting in total shoulder arthroplasty. AB - Abnormal glenoid architecture resulting from loss of bone usually is listed among the contraindications to total shoulder arthroplasty using an unconstrained prosthesis. However, in a series of 463 consecutive replacement procedures that were performed between 1973 and 1985, in only two patients did the lack of bone make the implantation of a glenoid component impossible. Of the remaining sixty five shoulders that had an abnormal glenoid, twenty were successfully treated with a large, internally fixed bone graft or grafts and forty-five, with smaller bone grafts that were not internally fixed. Nineteen of the twenty shoulders that had a large graft or grafts were followed for two years or more (average, 4.4 years). The clinical results were judged to be excellent in sixteen and satisfactory in one, and the desired limited goals were obtained in two. Two fixation screws broke and one screw was worn by contact with the humeral component. None of the glenoid components clinically loosened or migrated, and no patient has needed further surgical treatment. Although bone-grafting was necessary in only twenty (4.3 per cent) of the 463 replacement procedures, this procedure provided sufficient osseous support to allow implantation of a component in a severely damaged glenoid. PMID- 3047132 TI - Failure of bipolar hip arthroplasty secondary to retained antibiotic-impregnated polymethylmethacrylate beads. A case report. PMID- 3047133 TI - Current concepts review. Carpal instability. PMID- 3047134 TI - A modified technique for mitral valve replacement. AB - Between 1976 and 1983 254 mitral valve prostheses (243 Bjork-Shiley and 14 bioprostheses) were implanted in 252 patients using a modified technique. The age of patients varied from 26-72 years (mean 56 +/- 18 years). This technique allows an equidistant placement of sutures and accurate stitching of mitral valve annulus to the prosthetic valve ring. Follow-up averaged 67 +/- 21.6 months (3-7 years). Linearized aseptic dehiscence was 0.17 per 100 patient years. This technique was especially useful in patients with small left atrium and minimised the risk of a major prosthetic detachment. PMID- 3047135 TI - Repair of ventricular rupture following mitral valve replacement. AB - Left ventricular rupture is reported to occur in 0.5-2% of patients following mitral valve replacement and results in a high mortality rate. Three types of left ventricular rupture have been identified, each attributed to a different mechanism. Failure of repair has been due to repeated tearing of the ventricular muscle and resulting hemorrhage. We describe the repair of left ventricular rupture following mitral valve replacement with buttressed dacron patch. The repair is designed to eliminate the tension placed on the suture line. In addition, specific recommendations are made to avoid left ventricular rupture during mitral valve replacement. PMID- 3047136 TI - Surgical treatment of respiratory insufficiency due to tracheobronchial compression by aneurysms of the ascending aorta and innominate artery. AB - Two patients with life-threatening respiratory insufficiency resulting from tracheobronchial compression by expanding aneurysms of the ascending aorta and innominate artery were successfully treated by cardiopulmonary bypass, even in the presence of significant aortic regurgitation. The pertinent surgical procedures are described. PMID- 3047137 TI - Ruptured abdominal aortic aneurysm with fistula into the right iliac vein. AB - Arteriovenous fistula between major abdominal vessels is an unusual complication of ruptured abdominal aortic aneurysm. It most frequently occurs between the aorta and the inferior vena cava, however, it is rare when occurring between the aorta and iliac veins. A case of an arteriovenous fistula (AVF), secondary to erosion of an arteriosclerotic abdominal aortic aneurysm into both the retroperitoneum and the iliac vein is presented. The literature is reviewed and the symptoms and the treatment are discussed. PMID- 3047138 TI - Postnephrectomy arteriovenous fistula. AB - A new case of postnephrectomy arteriovenous fistula is reported. This is an uncommon acquired complication of a common surgical procedure. Clinical features, diagnostic procedures, surgical treatment and results are analysed and discussed. In this case, a direct communication between the right renal artery and the inferior vena cava was found. PMID- 3047139 TI - Human interleukin 1 beta is not secreted from hamster fibroblasts when expressed constitutively from a transfected cDNA. AB - To understand the secretion and processing of interleukin-1 (IL-1), a Chinese hamster fibroblast cell line (R1610) was transfected with a human IL-1 beta cDNA under the control of the SV40 early promoter and linked to the gene for neomycin resistance. After selecting for transfected cells resistant to G418, two clones were found to constitutively express the IL-1 beta 31-kD precursor which was almost exclusively located in the cytosol. Pulse-chase experiments failed to show any secretion of IL-1 and very little IL-1 activity was detectable in cell supernatants. Furthermore, surface membrane IL-1 activity could not be detected, although low levels of activity could be released upon brief trypsin treatment. Therefore, unlike monocytes, these fibroblast cells lack the mechanism for secreting and processing of IL-1 beta. PMID- 3047140 TI - Signals for the incorporation and orientation of cytochrome P450 in the endoplasmic reticulum membrane. AB - Cytochrome P450b is an integral membrane protein of the rat hepatocyte endoplasmic reticulum (ER) which is cotranslationally inserted into the membrane but remains largely exposed on its cytoplasmic surface. The extreme hydrophobicity of the amino-terminal portion of P450b suggests that it not only serves to initiate the cotranslational insertion of the nascent polypeptide but that it also halts translocation of downstream portions into the lumen of the ER and anchors the mature protein in the membrane. In an in vitro system, we studied the cotranslational insertion into ER membranes of the normal P450b polypeptide and of various deletion variants and chimeric proteins that contain portion of P450b linked to segments of pregrowth hormone or bovine opsin. The results directly established that the amino-terminal 20 residues of P450b function as a combined insertion-halt-transfer signal. Evidence was also obtained that suggests that during the early stages of insertion, this signal enters the membrane in a loop configuration since, when the amino-terminal hydrophobic segment was placed immediately before a signal peptide cleavage site, cleavage by the luminally located signal peptidase took place. After entering the membrane, the P450b signal, however, appeared to be capable of reorienting within the membrane since a bovine opsin peptide segment linked to the amino terminus of the signal became translocated into the microsomal lumen. It was also found that, in addition to the amino-terminal combined insertion-halt-transfer signal, only one other segment within the P450b polypeptide, located between residues 167 and 185, could serve as a halt-transfer signal and membrane-anchoring domain. This segment was shown to prevent translocation of downstream sequences when the amino-terminal combined signal was replaced by the conventional cleavable insertion signal of a secretory protein. PMID- 3047141 TI - Differential localization of tropomyosin isoforms in cultured nonmuscle cells. AB - We have previously shown that chicken embryo fibroblast (CEF) cells and human bladder carcinoma (EJ) cells contain multiple isoforms of tropomyosin, identified as a, b, 1, 2, and 3 in CEF cells and 1, 2, 3, 4, and 5 in human EJ cells by one dimensional SDS-PAGE (Lin, J. J.-C., D. M. Helfman, S. H. Hughes, and C.-S. Chou. 1985. J. Cell Biol. 100: 692-703; and Lin, J. J.-C., S. Yamashiro-Matsumura, and F. Matsumura. 1984. Cancer Cells 1:57-65). Both isoform 3 (TM-3) of CEF and isoforms 4,5 (TM-4,-5) of human EJ cells are the minor isoforms found respectively in normal chicken and human cells. They have a lower apparent molecular mass and show a weaker affinity to actin filaments when compared to the higher molecular mass isoforms. Using individual tropomyosin isoforms immobilized on nitrocellulose papers and sequential absorption of polyclonal antiserum on these papers, we have prepared antibodies specific to CEF TM-3 and to CEF TM-1, 2. In addition, two of our antitropomyosin mAbs, CG beta 6 and CG3, have now been demonstrated by Western blots, immunoprecipitation, and two-dimensional gel analysis to have specificities to human EJ TM-3 and TM-5, respectively. By using these isoform-specific reagents, we are able to compare the intracellular localizations of the lower and higher molecular mass isoforms in both CEF and human EJ cells. We have found that both lower and higher molecular mass isoforms of tropomyosin are localized along stress fibers of cells, as one would expect. However, the lower molecular mass isoforms are also distributed in regions near ruffling membranes. Further evidence for this different localization of different tropomyosin isoforms comes from double-label immunofluorescence microscopy on the same CEF cells with affinity-purified antibody against TM-3, and monoclonal CG beta 6 antibody against TM-a, -b, -1, and -2 of CEF tropomyosin. The presence of the lower molecular mass isoform of tropomyosin in ruffling membranes may indicate a novel way for the nonmuscle cell to control the stability and organization of microfilaments, and to regulate the cell motility. PMID- 3047144 TI - The centrin-based cytoskeleton of Chlamydomonas reinhardtii: distribution in interphase and mitotic cells. AB - Monoclonal and polyclonal antibodies raised against algal centrin, a protein of algal striated flagellar roots, were used to characterize the occurrence and distribution of this protein in interphase and mitotic Chlamydomonas cells. Chlamydomonas centrin, as identified by Western immunoblot procedures, is a low molecular (20,000-Mr) acidic protein. Immunofluorescence and immunogold labeling demonstrates that centrin is a component of the distal fiber. In addition, centrin-based flagellar roots link the flagellar apparatus to the nucleus. Two major descending fibers extend from the basal bodies toward the nucleus; each descending fiber branches several times giving rise to 8-16 fimbria which surround and embrace the nucleus. Immunogold labeling indicates that these fimbria are juxtaposed to the outer nuclear envelope. Earlier studies have demonstrated that the centrin-based linkage between the flagellar apparatus and the nucleus is contractile, both in vitro and in living Chlamydomonas cells (Wright, R. L., J. Salisbury, and J. Jarvik. 1985. J. Cell Biol. 101:1903-1912; Salisbury, J. L., M. A. Sanders, and L. Harpst. 1987. J. Cell Biol. 105:1799 1805). Immunofluorescence studies show dramatic changes in distribution of the centrin-based system during mitosis that include a transient contraction at preprophase; division, separation, and re-extension during prophase; and a second transient contraction at the metaphase/anaphase boundary. These observations suggest a fundamental role for centrin in motile events during mitosis. PMID- 3047143 TI - The mechanism of anaphase spindle elongation: uncoupling of tubulin incorporation and microtubule sliding during in vitro spindle reactivation. AB - To study tubulin polymerization and microtubule sliding during spindle elongation in vitro, we developed a method of uncoupling the two processes. When isolated diatom spindles were incubated with biotinylated tubulin (biot-tb) without ATP, biot-tb was incorporated into two regions flanking the zone of microtubule overlap, but the spindles did not elongate. After biot-tb was removed, spindle elongation was initiated by addition of ATP. The incorporated biot-tb was found in the midzone between the original half-spindles. The extent and rate of elongation were increased by preincubation in biot-tb. Serial section reconstruction of spindles elongating in tubulin and ATP showed that the average length of half-spindle microtubules increased due to growth of microtubules from the ends of native microtubules. The characteristic packing pattern between antiparallel microtubules was retained even in the "new" overlap region. Our results suggest that the forces required for spindle elongation are generated by enzymes in the overlap zone that mediate the sliding apart of antiparallel microtubules, and that tubulin polymerization does not contribute to force generation. Changes in the extent of microtubule overlap during spindle elongation were affected by tubulin and ATP concentration in the incubation medium. Spindles continued to elongate even after the overlap zone was composed entirely of newly polymerized microtubules, suggesting that the enzyme responsible for microtubule translocation either is bound to a matrix in the spindle midzone, or else can move on one microtubule toward the spindle midzone and push another microtubule of opposite polarity toward the pole. PMID- 3047142 TI - Analysis of the upstream regions governing expression of the chicken cardiac troponin T gene in embryonic cardiac and skeletal muscle cells. AB - The chicken gene encoding cardiac troponin T (cTNT) is expressed in both cardiac and skeletal muscle during early embryonic development, but is specifically repressed in skeletal muscle during fetal development. To determine if the cis acting sequences governing transcription of a single gene in these two related cell types are the same, we have transfected promoter/upstream segments of the cTNT gene coupled to the bacterial chloramphenicol acetyltransferase gene into primary cultures of early embryonic cardiac and skeletal muscle cells. Using this assay system, chloramphenicol acetyltransferase activity directed by the cTNT promoter/upstream region was between two and three orders of magnitude higher in cardiac or skeletal muscle cells than in fibroblast cells, indicating that cis elements responsible for cell-specific expression reside in this region of the cTNT gene. Deletion experiments showed that a 67-nucleotide DNA segment residing between 268 and 201 nucleotides upstream of the cTNT transcription initiation site is required for cTNT promoter activity in embryonic cardiac cells. This region is not required in embryonic skeletal muscle cells because a cTNT promoter construction containing only 129 upstream nucleotides is transcriptionally active in these cells. These results demonstrate that different cis-acting sequences are required for cTNT expression in early embryonic cardiac and skeletal muscle cells. Nonessential regions residing farther upstream, on the other hand, affected the level of expression of these minimum regions in a similar manner in both cell types. The data from these experiments indicate, therefore, that transcription of the cTNT promoter in early embryonic cardiac and skeletal muscle cells is governed both by common and divergent regulatory elements in cis and in trans. PMID- 3047145 TI - Microtubule dynamics in nerve cells: analysis using microinjection of biotinylated tubulin into PC12 cells. AB - To study microtubule (MT) dynamics in nerve cells, we microinjected biotin labeled tubulin into the cell body of chemically fused and differentiated PC12 cells and performed the immunofluorescence or immunogold procedure using an anti biotin antibody followed by secondary antibodies coupled to fluorescent dye or colloidal gold. Incorporation of labeled subunits into the cytoskeleton of neurites was observed within minutes after microinjection. Serial electron microscopic reconstruction revealed that existing MTs in PC12 neurites incorporated labeled subunits mainly at their distal ends and the elongation rate of labeled segments was estimated to be less than 0.3 micron/min. Overall organization of MTs in the nerve cells was different from that in undifferentiated cells such as fibroblasts. Namely, we have not identified any MT organizing centers from which labeled MTs are emanating in the cell bodies of the injected cells. Stereo electron microscopy revealed that some fully labeled segments seemed to start in the close vicinity of electron dense material within the neurites. This suggests new nucleation off some structures in the neurites. We have also studied the overall pattern of the incorporation of labeled subunits which extended progressively from the proximal part of the neurites toward their tips. To characterize the mechanism of tubulin incorporation, we have measured mean density of gold labeling per unit length of labeled segments at different parts of the neurites. The results indicate access of free tubulin subunits into the neurites and local incorporation into the neurite cytoskeleton. Our results lead to the conclusion that MTs are not static polymers but dynamic structures that continue to elongate even within the differentiated nerve cell processes. PMID- 3047146 TI - Distribution and role in regeneration of N-CAM in the basal laminae of muscle and Schwann cells. AB - The neural cell adhesion molecule (N-CAM) is a membrane glycoprotein involved in neuron-neuron and neuron-muscle adhesion. It can be synthesized in various forms by both nerve and muscle and it becomes concentrated at the motor endplate. Biochemical analysis of a frog muscle extract enriched in basal lamina revealed the presence of a polydisperse, polysialylated form of N-CAM with an average Mr of approximately 160,000 as determined by SDS-PAGE, which was converted to a form of 125,000 Mr by treatment with neuraminidase. To define further the role of N CAM in neuromuscular junction organization, we studied the distribution of N-CAM in an in vivo preparation of frog basal lamina sheaths obtained by inducing the degeneration of both nerve and muscle fibers. Immunoreactive material could be readily detected by anti-N-CAM antibodies in such basal lamina sheaths. Ultrastructural analysis using immunogold techniques revealed N-CAM in close association with the basal lamina sheaths, present in dense accumulation at places that presumably correspond to synaptic regions. N-CAM epitopes were also associated with collagen fibrils in the extracellular matrix. The ability of anti N-CAM antibodies to perturb nerve regeneration and reinnervation of the remaining basal lamina sheaths was then examined. In control animals, myelinating Schwann cells wrapped around the regenerated axon and reinnervation occurred only at the old synaptic areas; new contacts between nerve and basal lamina had a terminal Schwann cell capping the nerve terminal. In the presence of anti-N-CAM antibodies, three major abnormalities were observed in the regeneration and reinnervation processes: (a) regenerated axons in nerve trunks that had grown back into the old Schwann cell basal lamina were rarely associated with myelinating Schwann cell processes, (b) ectopic synapses were often present, and (c) many of the axon terminals lacked a terminal Schwann cell capping the nerve basal lamina contact area. These results suggest that N-CAM may play an important role not only in the determination of synaptic areas but also in Schwann cell axon interactions during nerve regeneration. PMID- 3047147 TI - Type VIII collagen has a restricted distribution in specialized extracellular matrices. AB - A pepsin-resistant triple helical domain (chain 50,000 Mr) of type VIII collagen was isolated from bovine corneal Descemet's membrane and used as an immunogen for the production of mAbs. An antibody was selected for biochemical and tissue immunofluorescence studies which reacted both with Descemet's membrane and with type VIII collagen 50,000-Mr polypeptides by competition ELISA and immunoblotting. This antibody exhibited no crossreactivity with collagen types I VI by competition ELISA. The mAb specifically precipitated a high molecular mass component of type VIII collagen (EC2, of chain 125,000 Mr) from the culture medium of subconfluent bovine corneal endothelial cells metabolically labeled for 24 h. In contrast, confluent cells in the presence of FCS and isotope for 7 d secreted a collagenous component of chain 60,000 Mr that did not react with the anti-type VIII collagen IgG. Type VIII collagen therefore appears to be synthesized as a discontinuous triple helical molecule with a predominant chain 125,000 Mr by subconfluent, proliferating cells in culture. Immunofluorescence studies with the mAb showed that type VIII collagen was deposited as fibrils in the extracellular matrix of corneal endothelial cells. In the fetal calf, type VIII collagen was absent from basement membranes and was found in a limited number of tissues. In addition to the linear staining pattern observed in the Descemet's membrane, type VIII collagen was found in highly fibrillar arrays in the ocular sclera, in the meninges surrounding brain, spinal cord, and optic nerve, and in periosteum and perichondrium. Fine fibrils were evident in the white matter of spinal cord, whereas a more generalized staining was apparent in the matrices of cartilage and bone. Despite attempts to unmask the epitope, type VIII collagen was not found in aorta, kidney, lung, liver, skin, and ligament. We conclude that this unusual collagen is a component of certain specialized extracellular matrices, several of which are derived from the neural crest. PMID- 3047149 TI - Studies on cardiac myofibrillogenesis with antibodies to titin, actin, tropomyosin, and myosin. AB - Cardiac myofibrillogenesis was examined in cultured chick cardiac cells by immunofluorescence using antibodies against titin, actin, tropomyosin, and myosin. Primitive cardiomyocytes initially contained stress fiber-like structures (SFLS) that stained positively for alpha actin and/or muscle tropomyosin. In some cases the staining for muscle tropomyosin and alpha actin was disproportionate; this suggests that the synthesis and/or assembly of these two isoforms into the SFLS may not be stoichiometric. The alpha actin containing SFLS in these myocytes could be classified as either central or peripheral; central SFLS showed developing sarcomeric titin while peripheral SFLS had weak titin fluorescence and a more uniform stain distribution. Sarcomeric patterns of titin and myosin were present at multiple sites on these structures. A pair of titin staining bands was clearly associated with each developing A band even at the two or three sarcomere stage, although occasional examples of a titin band being associated with a half sarcomere were noted. The appearance of sarcomeric titin patterns coincided or preceded sarcomere periodicity of either alpha actin or muscle tropomyosin. The early appearance of titin in myofibrillogenesis suggests it may have a role in filament alignment during sarcomere assembly. PMID- 3047148 TI - Laser-transected microtubules exhibit individuality of regrowth, however most free new ends of the microtubules are stable. AB - To study the possible mechanism of microtubule turnover in interphase cells, we have used the 266-nm wavelength of a short-pulsed Nd/YAG laser to transect microtubules in situ in PtK2 cells at predefined regions. The regrowth and shrinkage of the transected microtubules have been examined by staining the treated cells with antitubulin mAb at various time points after laser irradiation. The results demonstrate that microtubules grow back into the transected zones individually; neither simultaneous growth nor shrinkage of all microtubules has been observed. The half-time of replacement of laser-dissociated microtubules is observed to be approximately 10 min. On the other hand, exposure of the core of the microtubule, which is expected to consist almost completely of GDP-tubulin, by transecting the internal regions of the microtubule does not render the remaining polymer catastrophically disassembled, and most transected microtubules with free minus ends do not quickly disappear. Taken together, these results suggest that most microtubules in cultured interphase cells exhibit some properties of dynamic instability (individual regrowth or shrinkage); however, other factors in addition to the hydrolysis of GTP-tubulin need to be involved in modulating the dynamics and the stability of these cytoplasmic microtubules. PMID- 3047150 TI - Attachment to an endogenous laminin-like protein initiates sprouting by leech neurons. AB - Leech neurons in culture sprout rapidly when attached to extracts from connective tissue surrounding the nervous system. Laminin-like molecules that promote sprouting have now been isolated from this extracellular matrix. Two mAbs have been prepared that react on immunoblots with a approximately equal to 220- and a approximately equal to 340-kD polypeptide, respectively. These antibodies have been used to purify molecules with cross-shaped structures in the electron microscope. The molecules, of approximately equal to 10(3) kD on nonreducing SDS gels, have subunits of approximately equal to 340, 220, and 160-180 kD. Attachment to the laminin-like molecules was sufficient to initiate sprouting by single isolated leech neurons in defined medium. This demonstrates directly a function for a laminin-related invertebrate protein. The mAbs directed against the approximately equal to 220-kD chains of the laminin-like leech molecule labeled basement membrane extracellular matrix in leech ganglia and nerves. A polyclonal antiserum against the approximately equal to 220-kD polypeptide inhibited neurite outgrowth. Vertebrate laminin did not mediate the sprouting of leech neurons; similarly, the leech molecule was an inert substrate for vertebrate neurons. Although some traits of structure, function, and distribution are conserved between vertebrate laminin and the invertebrate molecule, our results suggest that the functional domains differ. PMID- 3047151 TI - A membrane glycoprotein, Sec12p, required for protein transport from the endoplasmic reticulum to the Golgi apparatus in yeast. AB - SEC12, a gene that is required for secretory, membrane, and vacuolar proteins to be transported from the endoplasmic reticulum to the Golgi apparatus, has been cloned from a genomic library by complementation of a sec12 ts mutation. Genetic analysis has shown that the cloned gene integrates at the SEC12 locus and that a null mutation at the locus is lethal. The DNA sequence predicts a protein of 471 amino acids containing a hydrophobic stretch of 19 amino acids near the COOH terminus. To characterize the gene product (Sec12p) in detail, a lacZ-SEC12 gene fusion has been constructed and a polyclonal antibody raised against the hybrid protein. The antibody recognizes Sec12p as a approximately 70-kD protein that sediments in a mixed membrane fraction that includes endoplasmic reticulum. Sec12p is not removed from the membrane fraction by treatment at high pH and high salt and is not degraded by exogenous protease unless detergent is present. Glycosylation of Sec12p during biogenesis is indicated by an electrophoretic mobility shift of the protein that is influenced by tunicamycin and by imposition of an independent secretory pathway block. We suggest that Sec12p is an integral membrane glycoprotein with a prominent domain that faces the cytoplasm where it functions to promote protein transport to the Golgi apparatus. In the process of transport, Sec12p itself may migrate to the Golgi apparatus and function in subsequent transport events. PMID- 3047152 TI - Cell surface expression of glycosylated, nonglycosylated, and truncated forms of a cytoplasmic protein pyruvate kinase. AB - The soluble cytoplasmic protein pyruvate kinase (PK) has been expressed at the cell surface in a membrane-anchored form (APK). The hybrid protein contains the NH2-terminal signal/anchor domain of a class II integral membrane protein (hemagglutinin/neuraminidase, of the paramyxovirus SV5) fused to the PK NH2 terminus. APK contains a cryptic site that is used for N-linked glycosylation but elimination of this site by site-specific mutagenesis does not prevent cell surface localization. Truncated forms of the APK molecule, with up to 80% of the PK region of APK removed, can also be expressed at the cell surface. These data suggest that neither the complete PK molecule nor its glycosylation are necessary for intracellular transport of PK to the cell surface, and it is possible that specific signals may not be needed in the ectodomain of this hybrid protein to specify cell surface localization. PMID- 3047155 TI - Somatotropin antagonism of insulin-stimulated glucose utilization in 3T3-L1 adipocytes. AB - It is well established that somatotropin (GH) antagonizes insulin action in vivo and that supraphysiologic concentrations of GH frequently result in insulin resistance and glucose intolerance. However, the demonstration of an anti-insulin activity by GH in vitro has been difficult. This study, therefore, set out to determine whether cultures of 3T3-L1 adipocytes could be used to examine the anti insulin activity of GH. The ability of insulin to stimulate glucose utilization by 3T3-L1 adipocytes increases approximately five-fold during the first 4 days following treatment of the cells with a differentiation medium. It was found that glucose utilization in 3T3-L1 adipocytes is regulated in a reciprocal fashion by insulin and GH. Bovine or human GH directly inhibit up to 50% of insulin stimulated [14C]-glucose incorporation into lipids in a concentration-dependent manner. The 3T3-L1 sensitivity to GH appears to be at the maximum (50% inhibition of an insulin response) immediately following removal of the cells from the differentiation medium and remains essentially constant during the subsequent 4 days. The GH inhibition of insulin action does not appear to be due GH enhancement of cellular degradation of insulin, competitive binding of GH to the insulin receptor, or GH-induced decrease in cell number. The 3T3-L1 adipocyte system appears to be a sensitive and reliable in vitro model with which to study the molecular mechanisms involved in both GH antagonism of insulin action and development of hormone responsiveness during cellular differentiation into adipocytes. PMID- 3047153 TI - Biogenesis of thylakoid membranes is controlled by light intensity in the conditional chlorophyll b-deficient CD3 mutant of wheat. AB - Biogenesis of thylakoid membranes in the conditional chlorophyll b-deficient CD3 mutant of wheat is dramatically altered by relatively small differences in the light intensity under which seedlings are grown. When the CD3 mutant is grown at 400 microE/m2 S (high light, about one-fifth full sunlight) plants are deficient in chlorophyll b (chlorophyll a/b ratio greater than 6.0) and lack or contain greatly reduced amounts of the chlorophyll a/b-binding complexes CPII/CPII (mobile or peripheral LHCII), CP29, CP24 and LHCI, as shown by mildly denaturing 'green gel' electrophoresis, by fully denaturing SDS-PAGE, and by Western blot analysis. High light CD3 chloroplasts display an unusual morphology characterized by large, sheet-like stromal thylakoids formed into parallel unstacked arrays and a limited number of small grana stacks displaced toward the edges of the arrays. Changes in the supramolecular organization of CD3 thylakoids, seen with freeze fracture electron microscopy, include a reduction in the size of EFs particles, which correspond to photosystem II centers with variable amounts of attached LHCII, and a redistribution of EF particles from the stacked to the unstacked regions. When CD3 seedlings are grown at 150 microE/m2 S (low light) there is a substantial reversal of all of these effects. Thus, chlorophyll b and the chlorophyll a/b-binding proteins accumulate to near wild-type levels (chlorophyll a/b ratio = 3.5-4.5) and thylakoid morphology is more nearly wild type in appearance. Growth of the CD3 mutant in the presence of chloramphenicol stimulates the accumulation of chlorophyll b and its binding proteins (Duysen, M. E., T. P. Freeman, N. D. Williams, and L. L. Huckle. 1985. Plant Physiol. 78:531 536). We show that this partial rescue of the CD3 high light phenotype is accompanied by large changes in thylakoid structure. The CD3 mutant, which defines a new class of chlorophyll b-deficient phenotype, is discussed in the more general context of chlorophyll b deficiency. PMID- 3047156 TI - Localization of platelet antigens and fibrinogen on osteoclasts. AB - The antigenic phenotype of the human osteoclast, which is known to be derived from a circulating mononuclear precursor cell of haemopoietic origin, is controversial. Recent studies have shown that macrophage as well as megakaryocyte/platelet antigens are expressed by osteoclasts. In this study, we have sought to define, by immunohistochemistry, the nature and possible function of platelet antigens expressed by human osteoclasts in foetal and adult bone specimens. Monoclonal antibodies to platelet glycoprotein IIIa (gpIIIa) and CD9 antibodies stained osteoclasts in all bone specimens examined. Fibrinogen was also localized to the osteoclast membrane in foetal bone imprints. In addition, we found that CD9 and gpIIIa antibodies reacted weakly with monocytes in buffy coat smears. Antibodies to factor 8 and glycoproteins Ib and IIb/IIIa did not react with osteoclasts. These results show that osteoclasts, monocytes, macrophages, megakaryocytes and platelets possess common antigens and that fibrinogen is present on the surface of osteoclasts. By analogy with platelets, CD9 and gpIIIa may play a role in fibrinogen binding by osteoclasts. Possible mechanisms by which platelet antigens and fibrinogen binding could affect osteoclast function are proposed. PMID- 3047158 TI - Structure of myofibrils at extra-junctional membrane attachment sites in cultured cardiac muscle cells. AB - The composition and organization of myofibrils at extra-junctional membrane attachment sites in cultured neonatal rat cardiac muscle cells were analysed by immunofluorescence and electron microscopy. When myofibril terminals attached to the cell membrane via focal contacts at regions of the sarcolemma that lacked intercalated discs, they appeared to be non-striated and resembled thick actin cables. Although the non-striated terminals contained actin, myosin and alpha actinin, the proteins were not organized into recognizable sarcomeres at the light microscopic level. Analysis of the structure of the terminals in the electron microscope confirmed that the usual sarcomeric organization and attachments to the sarcolemma were markedly modified. The non-striated myofibril terminals differed in structure from both stress fibres in non-muscle cells and stress fibre-like structures present in embryonic heart cells in culture. Non striated myofibril terminals attached to the cell membrane by lateral contact with extra-junctional electron-dense membrane plaques rather than by insertion by their ends into the fascia adherens. It is proposed that the structure and composition of membrane-attachment points for myofibrils may have an influence on the structure, organization or stability of contractile elements in cardiac muscle. PMID- 3047157 TI - Distribution of pericentriolar material in multipolar spindles induced by colcemid treatment in Chinese hamster ovary cells. AB - Mitotic Chinese hamster ovary cells were obtained by treatment with microtubule drugs under various conditions, and the shape of spindles was analysed by phase contrast microscopy of isolated spindles, and by indirect immunofluorescence staining of whole mitotic cells with anti-tubulin antibody. Bipolarity of spindles was maintained after treatment with 0.05 microM of colcemid for 3.5 h, but increased exposure to higher concentrations (0.32 microM) and for longer durations (5.5 h) led to a marked rise in multipolar spindles. Nocodazole treatment, on the other hand, failed to show a multiplicity of spindle poles even at 3.3 microM. Each pole of a multipolar spindle was associated with pericentriolar material, as shown by staining with an autoimmune serum specific for pericentriolar material. The number of locations with free pericentriolar material capable of polymerizing microtubules in vitro also increased with increasing numbers of spindle poles, suggesting that dispersion of the pericentriolar material resulted in the production of many microtubule-nucleating sites in multipolar spindles. The different efficiencies of recovery from different drugs, which have been known to be quite variable, may be partly due to the different extent of dispersion of the pericentriolar material. PMID- 3047154 TI - Immunocytochemical evidence for translocation of protein kinase C in human megakaryoblastic leukemic cells: synergistic effects of Ca2+ and activators of protein kinase C on the plasma membrane association. AB - Immunological analysis using monoclonal antibodies against subspecies of protein kinase C revealed the predominant expression of the isozyme, type II, in human megakaryoblastic leukemic cells. We investigated the effects of phorbol diester 12-O-tetradecanoyl phorbol-13-acetate (TPA), the Ca2+ ionophore ionomycin and synthetic diacylglycerol 1-oleoyl-2-acetylglycerol (OAG) on the immunocytochemical localization of protein kinase C in these cells. Indirect immunofluorescence techniques revealed the enzyme to be located in a diffuse cytosolic pattern, in the intact cells. When the cells were exposed to 100 nM TPA, the immunofluorescent staining was translocated from the cytoplasm to the plasma membrane. The translocation was protracted and staining on the membrane decreased in parallel with the Ca2+, phospholipid-dependent protein kinase activity. Treatment of the cells with 500 nM ionomycin caused an apparent translocation comparable with that seen with TPA, however, this translocation was transient and most of the cytosolic staining was within 60 min. We also found that 30 micrograms/ml OAG did not have significant effects on distribution of the staining, but rather acted synergistically on the translocation with the suboptimal concentration of 100 nM ionomycin. A similar synergism was also observed with 10 nM TPA and 100 nM ionomycin. These results obtained in situ provide evidence that intracellular Ca2+ and diacylglycerol regulate membrane binding of the enzyme in vivo. PMID- 3047159 TI - Asperger's syndrome. PMID- 3047160 TI - A preliminary evaluation of five HIV antigen detection assays. AB - Five commercial assays for detecting HIV antigen were evaluated using a panel of 40 coded samples in six laboratories. All assays were capable of detecting HIV-1 antigen (HIV-1 Ag), and are likely to prove useful for monitoring supernatant fluids from a variety of cell cultures. The concentration of HIV-1 protein which the assays detected varied from 25 ng/ml down to 25 pg/ml. Three of the five assays were also able to detect HIV-2 Ag. More extensive evaluations are needed to determine the sensitivity and specificity of these assays on serum samples and body fluids. PMID- 3047161 TI - Insulin and glucagon secretory responses to arginine, glucagon, and theophylline during perifusion of human fetal islet-like cell clusters. AB - The secretory responsiveness of human fetal pancreatic endocrine cells was studied by perifusion of cultured islet-like cell clusters (ICC). ICC were obtained from 7 fetuses at 13-15 weeks gestation and 21 fetuses at 17-22 weeks gestation. The ICC were challenged with glucose (20 mmol/L), arginine (10 mmol/L), glucagon (1.4 mumol/L), and theophylline (10 mmol/L) combined with zero, low (2 mmol/L), or high (20 mmol/L) glucose. At 13-15 weeks, glucose and arginine enhanced insulin release in some experiments, whereas glucagon and theophylline were always potent stimuli (mean response, 4-fold regardless of the glucose concentration). At 17-22 weeks, both glucose alone (20 mmol/L) and arginine (10 mmol/L, with 2 mmol/L glucose) induced a small (1.4- to 1.5-fold) increase in insulin release. When arginine was combined with 20 mmol/L glucose, the response was potentiated to become a 2.3-fold increase. In contrast, glucagon was equally effective in 2 and 20 mmol/L glucose (2.9- and 2.6-fold responses, respectively) and produced a half-maximal response even in the absence of glucose. In this age range the most potent stimulus for insulin release was clearly theophylline. The effect of theophylline was also remarkably independent of the glucose concentration of the perifusate (5.6-, 8.1-, and 8.6-fold responses at 0, 2, and 20 mmol/L glucose, respectively). Glucagon release from the ICC of the 17- to 22 week-old fetuses was low (mean basal glucagon release, 2.9; insulin, 24.8 fmol/100 ICC/min). Glucagon release was not affected by 20 mmol/L glucose, but was stimulated by arginine and theophylline. These findings suggest that in the fetal pancreas, in contrast to the adult organ, insulin release results from elevation of intracellular cAMP concentrations (by glucagon or theophylline) relatively independent of the exogenous glucose concentration. Therefore, glucagon may have an important role in regulating insulin release during the early development of human fetal B-cells. PMID- 3047162 TI - Glucose metabolism in obesity: influence of body fat distribution. AB - The dose-response relationships between portal venous insulin concentrations and hepatic glucose production and between peripheral insulin concentrations and peripheral glucose utilization were determined in 8 nonobese and 17 obese premenopausal women with either upper or lower body fat localization. The glucose production dose-response curves for the two obese groups were shifted to the right at all levels of portal insulinemia. The upper body obese women had a greater rightward shift compared to the lower body obese women. The peripheral glucose utilization dose-response curve was shifted to the right in the lower body obese women, but maximal glucose utilization was normal. The upper body obese women had both a greater rightward shift and a marked reduction in maximal glucose utilization. The insulin concentrations that had half-maximal effects on glucose production and utilization were similar in each group. These results indicate that the liver is not inherently more sensitive to insulin than peripheral tissues. Obesity is associated with a moderate diminution of hepatic and peripheral insulin sensitivity. Upper body fat localization in obese women is characterized by a greater diminution in insulin sensitivity and decline in peripheral insulin responsivity than is lower body fat localization. The marked peripheral insulin resistance in the former group may account for the increased prevalence of glucose intolerance. PMID- 3047164 TI - Serum parathyroid hormone (PTH) in pregnant women determined by an immunoradiometric assay for intact PTH. AB - Most studies of circulating PTH levels using traditional RIAs have supported the concept of physiological hyperparathyroidism of pregnancy, with pregnant women having serum immunoreactive PTH levels significantly higher than those in nonpregnant subjects. However, such RIAs are insensitive and often detect inactive PTH fragments, so that the correlation between PTH immunoreactivity and bioactivity is poor. Employing a new intact PTH immunoradiometric assay (Allegro Nichols), we reassessed the effects of pregnancy on parathyroid function. The mean serum PTH level in 81 pregnant women was 14.4 +/- 6.3 (+/- SD) compared to 24.8 +/- 9.0 ng/L in 11 normally cycling nonpregnant women (P less than 0.001). The mean serum total and ionized calcium levels in the 2 groups were similar. In 5 of the pregnant women, serum bioactive PTH, determined by cytochemical bioassay, was slightly lower (7.7 +/- 3.4 ng/L) than in normal individuals (11.1 +/- 1.9 ng/L). Our findings suggest, in contrast with the results of most previous studies, that serum intact PTH may decline during pregnancy. PMID- 3047165 TI - Consumers in a competition-based cost containment environment. PMID- 3047163 TI - Defects in beta-cell function and insulin sensitivity in normoglycemic streptozocin-treated baboons: a model of preclinical insulin-dependent diabetes. AB - During the preclinical period of human insulin-dependent diabetes, both impaired pancreatic beta-cell function and increased insulin resistance are found, although normoglycemia is preserved. To better understand the changes in beta cell function and insulin sensitivity that occur in preclinical insulin-dependent diabetes, we performed a panel of in vivo beta-cell function tests and measured insulin sensitivity in adolescent male baboons both in normal health and after a small dose of streptozocin which did not induce hyperglycemia. Nine animals were studied before (stage 1) and 1 week after receiving a low dose of streptozocin (stage 2). There was no change in fasting plasma glucose or insulin. The mean glucose disposal rate (Kg) remained within the normal range, but dropped from 2.0 +/- 0.2% +/- SE) to 1.2 +/- 0.1%/min (P less than 0.01), the acute insulin response to arginine (AIR(arg)) fell from 67.7 +/- 19.4 microU/mL (485.8 +/- 139.2 pmol/L) to 32.8 +/- 7.2 microU/mL (235.3 +/- 51.7 pmol/L; P less than 0.05), and the acute insulin response to glucose (AIR(gluc)) fell from 881 +/- 243 microU/mL.10 min (6321 +/- 1744 pmol/L.10 min) to 334 +/- 82 microU/mL.10 min (2396 +/- 588 pmol/L.10 min; P less than 0.01). The most dramatic change, however, was in the ability of hyperglycemia to potentiate AIR(arg) (expressed as the slope of potentiation). This was reduced by 94% from 1.8 +/- 0.5 to 0.1 +/- 0.1 (P less than 0.01), with almost no overlap in values between stages 1 and 2. Insulin sensitivity was also lower 1 week after streptozocin treatment. When the animals were restudied 8 weeks after streptozocin treatment (stage 3) most measures of beta-cell function were not significantly different from those in stage 1. The fasting plasma glucose level was 85.4 +/- 4.3 mg/dL (4.7 +/- 0.2 mmol/L), Kg was 1.8 +/- 0.3%/min, fasting plasma insulin was 35.9 +/- 8.5 microU/mL (257.6 +/- 61.0 pmol/L), AIR(arg) was 67.0 +/- 15.4 microU/mL (480.7 +/ 110.5 pmol/L), and AIR(gluc) was 615.3 +/- 265.3 microU/mL.10 min (4413 +/- 1901 pmol/L.10 min), and tissue insulin sensitivity was 2.7 +/- 0.4 x 10(4) min/microU.mL. These values show extensive overlap with those of stage 1, from which they are not significantly different. The slope of glucose potentiation, however, remained low in all animals at stage 3.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3047166 TI - Characterization of a human hematopoietic progenitor cell capable of forming blast cell containing colonies in vitro. AB - A hematopoietic cell (CFU-B1) capable of producing blast cell containing colonies in vitro was detected using a semisolid culture system. The CFU-B1 has the capacity for self-renewal and commitment to a number of hematopoietic lineages. Monoclonal antibody to the human progenitor cell antigen-1 (HPCA-1) and a monoclonal antibody against the major histocompatibility class II antigen (HLA DR) were used with fluorescence activated cell sorting to phenotype the CFU-B1. The CFU-B1 was found to express My10 but not HLA-DR antigen; experiments using complement-dependent cytotoxicity to eliminate DR positive cells confirmed this finding. Pretreatment of marrow cells with two chemotherapeutic agents, 5 fluorouracil and 4-hydroperoxycyclophosphamide facilitated detection of CFU-B1 derived colonies, while diminishing or totally inhibiting colony formation by other hematopoietic progenitor cells. CFU-B1-derived colony formation was dependent upon the addition of exogenous hematopoietic growth factors. Media conditioned either by the human bladder carcinoma cell line 5637 or lectin stimulated leukocytes, as well as recombinant granulocyte-macrophage colony stimulating factor, interleukin 3 or interleukin 1 alpha promoted blast cell colony formation. By contrast, neither recombinant erythropoietin, recombinant interleukin 4, purified macrophage colony stimulating factor or recombinant granulocyte colony-stimulating factor alone promoted blast cell colony formation. PMID- 3047168 TI - Growth hormone, 1988. PMID- 3047167 TI - In situ analysis of pancreatic islets in rats developing diabetes. Appearance of nonendocrine cells with surface MHC class II antigens and cytoplasmic insulin immunoreactivity. AB - Aberrant expression of MHC class II molecules on endocrine cells has been proposed to induce autoimmune reactions in thyroid and endocrine pancreas. The present study examines whether MHC class II positive insulin-containing islet cells occur at the onset of diabetes in rats, in analogy to the findings in man. At the onset of diabetes, both streptozotocin-treated and diabetes-prone BB rats exhibited numerous class II positive islet cells that presented ultrastructural features of monocytes and were surrounded by class II negative islet B cells. These class II positive cells were characterized by vacuoles that contained insulin immunoreactive granules and disrupted membranes. Similar cells also appeared positive for the monocyte marker OX-42. The presence of class II positive monocytes with insulin-containing vacuoles may indicate a removal of damage B cells by infiltrating leukocytes. A similar electron microscopical study in man will be necessary to distinguish the putative endocrine pancreatic B cells with aberrant class II expression from infiltrating nonendocrine class II positive cells with insulin-containing phagosomes. PMID- 3047170 TI - Serum prostacyclin stabilizing factor is identical to apolipoprotein A-I (Apo A I). A novel function of Apo A-I. AB - Serum PGI2 stabilizing factor (PSF) was purified from human serum to a single protein with a molecular weight of 28,000 D by SDS-PAGE. Analyses of NH2-terminal sequence (32 residues), COOH-terminal sequence (3 residues) and the composition of amino acids disclosed its homology with human apolipoprotein A-I (Apo A-I), a major apolipoprotein of HDL. Apolipoprotein A-II, C-I, C-II, C-III, D and E, as well as LDL, and VLDL did not possess this activity. The alpha-helix structure of Apo A-I is necessary for the binding of PGI2. HDL and nascent HDL reconstituted from Apo A-I and phospholipid significantly prolonged the half-life of PGI2. PGI2 stabilization by HDL and Apo A-I may be an important protective action against the accumulation of platelet thrombi at sites of vascular damage. The beneficial effect of HDL in the prevention of coronary artery disease may be partly due to this action. PMID- 3047169 TI - Predominant expression of circulating CD3+ lymphocytes bearing gamma T cell receptor in a prolonged immunodeficiency after allogeneic bone marrow transplantation. AB - The cell surface expression of alpha:beta heterodimer was studied using WT31 monoclonal antibody, in peripheral blood lymphocytes (PBL) from a patient who developed a prolonged immunodeficiency after allogeneic bone marrow transplantation. This patient, grafted for chronic myelogenous leukemia, received T cell depleted bone marrow from her HLA, A, B, D matched sibling. The late occurrence of opportunistic infection, led us to analyze the phenotype of patient PBL. 70% of PBL were CD3+ and 29% WT31+, indicating that the majority of CD3+ PBL did not express the alpha:beta heterodimer. Transcription of the genes encoding the alpha, beta, and gamma chains was assessed in cell lines derived from PBL, by Northern blot analysis. We showed that the CD3+ WT31- subset expressed a truncated, beta mRNA (1.0 kb) and also truncated alpha transcript (1.4 kb). To determine the CD3-associated structure on CD3+ WT31- cell line, immunoprecipitation assays were performed using monoclonal anti-CD3 and an hetero antiserum against gamma peptides. These CD3+ WT31- cells expressed a disulfide linked dimer, composed of products of gamma gene (37 kD, 40 kD) and of undefined delta chain (45 kD). Functional analyses were performed in PBL before and after sorting with WT31 and anti-CD3 antibody. These circulating CD3+ WT31- cells were unable to proliferate when triggered with anti-T3 beads and they seemed to mediate a suppressor activity on CD3+ WT31+ cells. PMID- 3047173 TI - Application of statistical moment theory to pharmacokinetics. PMID- 3047174 TI - The Zimbabwe drug regulatory process: a review of aspects of its development and current operational status. PMID- 3047172 TI - Analysis of renal fibrosis in a rabbit model of crescentic nephritis. AB - The pathogenesis of renal fibrosis in crescentic nephritis is incompletely understood. To improve our understanding of this process, crescentic nephritis was induced in New Zealand White rabbits by administration of guinea pig antiglomerular basement membrane IgG after sensitization with guinea pig IgG, and their kidneys were analyzed for the development of fibrosis. Collagen synthesis in renal cortical tissue was significantly elevated by day 3, peaked at days 7 15, and returned towards baseline by day 21. Collagen content of both glomeruli and cortex were increased starting on days 14-16, and remained constant in cortex thereafter. Light microscopic analysis was much less sensitive, revealing fibrosis only after day 21. Immunofluorescence revealed that type IV collagen was distributed primarily in the glomerulus, while types I and III were increased in the glomerulus and interstitium. Thus, in this model of crescentic nephritis, fibrosis, as assessed biochemically, developed early at time points when morphologic analysis failed to detect such a development. Hence early therapeutic intervention, before morphologic evidence of fibrosis is evident, may be more successful in arresting the progression of this disease before it reaches irreversible terminal stages. PMID- 3047171 TI - Modulation of human platelet protein kinase C by endotoxic lipid A. AB - Lipid A is the toxic principle of lipopolysaccharide of gram-negative bacteria, which causes a spectrum of changes in blood cells and vascular cells. We now report that human platelets are directly stimulated by endotoxic lipid A that activates protein kinase C. Rapid phosphorylation of a human platelet protein of Mr 47,000, a marker of protein kinase C activation, accompanies secretion of [14C]serotonin and aggregation triggered by endotoxic lipid A. These events are time and concentration dependent, with phosphorylation reaching maximum in 2 min and the concentration of lipid A causing a 50% effect (EC50) between 12 and 15 microM. Phospholipase C activation in lipid A-stimulated platelets was not observed as judged by a lack of generation of [3H]diacylglycerol in [3H]arachidonic acid-labeled platelets and a lack of generation of [32P] phosphatidic acid in 32PO4-labeled platelets. Lipid A did not induce formation of TXA2 as measured by radioimmunoassay for TXB2. The stimulation of human platelets and activation of protein kinase C by endotoxic lipid A was blocked by lipid X, a structural precursor of lipid A. Lipid X also blocked the stimulation of human platelets by phorbol 12-myristate 13-acetate, suggesting that lipid A, lipid X and phorbol ester share reactive site(s) on the human platelet membrane. Although lipid X inhibited thrombin-induced phosphorylation of P47 it did not suppress secretion of [14C]serotonin, indicating the role of protein kinase C-independent pathways in platelet stimulation by thrombin. The inhibitory effect of lipid X did not involve generation of cyclic AMP in human platelet membrane preparations. These results indicate that human platelets are stimulated by endotoxic lipid A, a naturally occurring biologic modifier of protein kinase C. Due to the widespread presence of this enzyme in blood cells, vascular cells, and neurons, its modulation by lipid A may represent a significant mechanism underlying hematologic and circulatory derangements observed in endotoxic shock in humans. PMID- 3047175 TI - Drug-binding proteins in liver transplant patients. PMID- 3047177 TI - Clinical pharmacology of NSAIDs. AB - All nonsteroidal anti inflammatory drugs (NSAIDs) are characterized by a high degree of protein binding and small volumes of distribution. Differences in clearance account for the variability in half-life among these drugs. The majority are metabolized by the liver through a variety of pathways. These drugs are subject to drug interactions of several mechanisms, including protein-binding displacement interactions, induction or inhibition of hepatic drug metabolism, and competition for active renal tubular secretion with other organic acids. NSAIDs are also subject to special pharmacokinetic considerations about which a great deal has been learned recently. These include data demonstrating that assessment of disposition of these drugs using only total drug concentrations can be misleading and that one must instead assess the disposition of unbound, pharmacologically active drug. Second, it appears that only one enantiomer of the propionic acid NSAIDs is active; studies of the pharmacokinetics of this class of NSAID should include assessment of the active stereoisomer. Finally, the propionic acid NSAIDs are metabolized to acyl-glucuronide conjugates, which are unstable and can cleave back to the parent drug. This feature allows the paradox of a drug that is metabolized by the liver being able to accumulate in patients who have renal insufficiency. Because of these newer pharmacokinetic considerations, much of the old data concerning the clinical pharmacology of NSAIDs are obsolete. More precise new information is needed in this area. PMID- 3047178 TI - Nonsteroidal anti-inflammatory drugs and renal function. PMID- 3047176 TI - NSAIDs: an overview. PMID- 3047180 TI - Comments on possible long-term consequences of nonsteroidal anti-inflammatory use. PMID- 3047182 TI - Nonsteroidal anti-inflammatory drugs in the management of juvenile arthritis. PMID- 3047179 TI - Leukotrienes: regulation of biosynthesis, metabolism, and bioactivity. PMID- 3047181 TI - The gastrointestinal side effects of the nonsteroidal anti-inflammatory drugs. PMID- 3047183 TI - Drug metabolism in the elderly. PMID- 3047184 TI - Counting sectioned cells via mathematical reconstruction. AB - A new method for determining the number of neurons in sectioned tissue is presented. The method does not involve identification of subcellular structures; rather, it uses estimates of the mean diameters of sections of the neuronal somata (with or without nuclei). All such sections are termed profiles. A mathematical model is developed to reconstruct the cell population from a size histogram of the profiles. Although the model is simple, the calculations are numerous and best done on a computer. A program that performs these calculations is provided. We discuss the idealizations on which the model is based and test the method in various ways: on hand- and computer-generated data in which imaginary spheres of known size were sectioned; on two small samples of real cells for which both cell and profile size histograms were available; and on a sample of potatoes, sliced by hand. In every case the estimate was within 10% of the actual number of cells (or potatoes). The method is robust in that it is relatively insensitive to section thickness, sample size, somal morphology, and observer error with respect to missing the small or thin profiles from any given cell. Results from the present model are compared to those obtained by using other cell count correction schemes that are currently employed. We call our method recursive translation. PMID- 3047185 TI - Distribution of calcitonin gene-related peptide immunoreactivity in relation to the rat central somatosensory projection. AB - The distribution of the neuropeptide calcitonin gene-related peptide (CGRP) was studied in relation to the known subcortical somatosensory pathways and contiguous systems in the central nervous system (CNS) of rats by using peroxidase histochemical methods in order to relate zones of immunoreactivity (IR) to cytoarchitecture. CGRP is the most ubiquitous peptide found to date in sensory ganglion cells: principally small and medium-size neurons emitting thin axons inferred to be largely nociceptive in function on the basis of the peripheral distribution of their terminals. Its apparent absence in sympathetic axons provides an especially useful sensory marker. The distribution of CGRP-IR axons displays remarkable selectivity at each level of the CNS. The trigeminal root distributes axons primarily to the pericornual layers (laminae I and II) of spinal V nucleus caudalis and to subnucleus oralis, evading the subnucleus interpolaris and contributing only few axons to principal V. Although there are only a few CGRP-IR somata at each level, heavily labeled axon trajectories can be traced to the nuclei of the solitary tract, the parabrachial nuclei, several sectors of the caudal medial thalamus, and the central nucleus of the amygdala. A sector of labeled neuron somata lies contiguous to each of these axon terminal zones, the largest of which is a thalamic nucleus containing cells of distinctive dendritic architecture extending from the periaqueductal gray across the posterior group nuclei to the peripeduncular nucleus, forming a linear array at the mesodiencephalic junction. The relation of CGRP-IR axonal distribution to spinothalamic, visceral, and gustatory systems is discussed in the context of a specialized "chemosensory" component of the thin-fiber somatosensory system. PMID- 3047187 TI - Early development of serotonin-containing neurons and pathways as seen in wholemount preparations of the fetal rat brain. AB - The early development of serotonin-containing neurons was studied in wholemounts of the fetal rat brain (E12-E18). The wholemounts were treated immunocytochemically according to an immunoperoxidase technique to reveal a panorama of developing serotoninergic neurons. Serotoninergic neurons were localized to two discrete groups or clusters within the brainstem. Serotonin containing neurons were identified first at E12 forming a rostral cluster of cells just caudal to the mesencephalic flexure. The more caudal cluster of cells first appeared at E14 in the medulla. During the period from E12 to E18, the immunoreactive cells increased in number and acquired a more complex dendritic tree while migrating to their permanent position. At E16, cells of the rostral group exhibited remarkably uniform mediolateral orientation. The rostral group of immunoreactive neurons gave rise to almost all ascending fibers, whereas the caudal group gave rise to the majority of descending fibers. Growing serotoninergic fibers were tipped by prominent growth cones, which were strongly immunoreactive. The fibers demonstrated prominent orientational selectivity with an almost total separation into ascending and descending bundles. Some of the ascending immunoreactive fibers displayed acute changes in their direction of growth, suggesting that the orientation of serotininergic fibers is mediated by directional cues that are specific to particular populations of serotoninergic fibers. Serotoninergic axons within the medial forebrain bundle were demonstrated particularly well and their ascent and rate of growth toward the forebrain could be easily followed. Immunoreactive fibers entered the telencephalon at E17 two portals, one along the lateral border of the hypothalamus and one rostrally, adjacent to the olfactory tubercle. In wholemounts at E18, fibers arising from this latter location could be followed as two distinct bands within the pallium; a basal band located ventrolaterally, adjacent to the lateral olfactory tract, and a dorsal band located at the medial edge of the telencephalon. Both fascicles were directed toward the occipital pole and contained unbranched fibers. At E18, serotoninergic axons arising from these two loosely organized fascicles covered most of the frontal telencephalon. The results of the present study indicate that wholemounts of embryonic brain can provide novel spaciotemporal data on the development of neuro-transmitter systems and may in the future prove to be useful experimental preparations in developmental neurobiology. PMID- 3047186 TI - Immunohistochemical localization of gonadotropin-releasing-hormone-associated peptide in the brain of the frog. AB - Gonadotropin-releasing-hormone (GnRH)-associated peptide (GnRH-AP) is a 56 amino acid neuropeptide derived from the GnRH prohormone. GnRH-AP corresponds to the C terminal fragment flanking the GnRH peptide. Using an antiserum raised against human GnRH-AP [1-56], or against human GnRH, we have investigated the neuronal systems containing either peptide in the central nervous system of the frog Rana ridibunda by immunohistological techniques. A main group of GnRH-AP-containing perikarya was found in a dorsoventral orientation of the supra anterior preoptic area (SAPA) just in front of the preoptic recess. Fibers originating from these perikarya projected rostrally toward the medial septal nucleus and the diagonal band of Broca. A network of GnRH-AP-immunoreactive (ir) fibers runs caudally from the SAPA toward the ventral hypothalamus. A high density of GnRH-AP-ir terminals was found in the median eminence. A few positive fibers were detected in the neural lobe of the pituitary, particularly in the region bordering the pars intermedia. Labelling of consecutive sections by either GnRH-AP or GnRH antibodies showed that GnRH and GnRH-AP-like irs were contained in the same cells of the SAPA. The double-staining technique with electrophoretic elution confirmed the colocalization of GnRH and GnRH-AP within the same neurons. Such a coexistence indicates that frog GnRH originates from a high molecular weight precursor which is closely related to rat pro-GnRH. The relative preservation of the C-terminal sequence of the pro-GnRH during evolution suggests that GnRH-AP may possess intrinsic biological activity. The high density of GnRH-AP-containing neurons projecting through the external zone of the median eminence would support the concept that GnRH-AP is involved in the modulation of pituitary hormone secretion. PMID- 3047188 TI - Serum enzymes in the diagnosis of disease in man and animals. PMID- 3047189 TI - Political savvy or lost opportunity? Evolution of the American Nurses' Association policy for nursing education funding, 1952 to 1972. PMID- 3047190 TI - The Presidents. Joseph P. Cappuccio 1978-1979. PMID- 3047191 TI - Regional myocardial volume alterations induced by brief repeated coronary occlusion in conscious dogs. AB - The purpose of this study was to evaluate whether brief repeated coronary occlusions induce changes in regional myocardial geometry at rest. Five conscious dogs were instrumented for the measurement of subendocardial segment length and transmural wall thickness in the ischemic area, subendocardial segment length in the normally perfused area, coronary flow and left ventricular pressure. After recovery from surgery, 180 (mean) 2 min coronary occlusions were given over a period of 20 days. The heart rate at rest, left ventricular peak systolic and end diastolic pressures and peak positive first derivative of left ventricular pressure (dP/dt) remained unchanged throughout the experiment. In the normal area, the end-diastolic segment length at rest did not change significantly. By contrast, in the ischemic area, at 14 days after the initiation of repeated coronary occlusion, the end-diastolic regional cross-sectional area (product of segment length and wall thickness) at rest had increased by 9.7% (p less than 0.05); thereafter it decreased to 6.5% (p less than 0.05) above the value at rest before repeated occlusion despite an additional 6 days of coronary occlusions. At 10 days after the interruption of repeated occlusion, this value had regressed to 4.3% (p = NS) above control. These findings suggest the occurrence of regional myocardial hypertrophy confined to the ischemic area in response to the periodic ischemic stimulus. PMID- 3047193 TI - The origin of common cardiac defects. PMID- 3047192 TI - Evolutionary origin of cardiac malformations. AB - The author has proposed in previous publications that isolated cardiac malformations have an evolutionary origin. This is partly supported by the fact that isolated cardiac malformations found in humans occur also in other placental mammals as well as in birds. External gross examination of the heart in just over 5,000 birds was carried out during a 3 year period. Anomalies included one instance of duplicate hearts, two specimens in which no heart could be identified and in a fourth, a yellow-rumped warbler, the heart lay in the neck outside of the thoracic cavity. Published reports of similar occurrences of an ectopically placed heart concern birds, cattle and humans. The fact that various species of both placental mammals and birds show evidence of heritability for heart defects, and that these species cannot interbreed, combined with the fact that birds and mammals have many similar malformations, points to either a common external causative factor or a common origin. Genes that code the malformed heart must be transmitted with that part of the genetic makeup common to all birds and mammals. Malformations caused by teratogens produce widespread organ injury to a potentially normal embryo whereas the evolutionary malformation is an organ specific anomaly in an otherwise normal mammal or bird and occurs in widely separated species. The implications of this theory are important for parents of children with an isolated congenital heart defect who may have ingested one or another drug or chemical or have been exposed to toxins or infectious agents before or after conception of the affected offspring. PMID- 3047194 TI - Training programs in the United States in adult cardiology, pediatric cardiology and cardiothoracic surgery. The American College of Cardiology. PMID- 3047195 TI - Ethical activism. PMID- 3047196 TI - Sensitivity and specificity of intracoronary injection of acetylcholine for the induction of coronary artery spasm. AB - Intracoronary injection of acetylcholine has been shown to induce coronary spasm in patients with variant angina. To examine its sensitivity and specificity, incremental doses of acetylcholine (20, 50 and 100 micrograms into the left coronary artery and 20 and 50 micrograms into the right coronary artery) were injected into the coronary artery or arteries in 70 patients with variant angina (Group 1) (mean age 57 years) and 93 patients without variant angina or angina at rest (Group 2) (mean age 54 years). Forty patients of the latter group had atypical chest pain, 16 cardiomyopathy, 14 arrhythmia, 11 valvular disease, 7 stable effort angina due to advanced coronary artery disease, 3 congenital heart disease and 2 hypertension. A temporary cardiac pacemaker set at 40 to 50 beats/min was positioned in the right ventricle. Coronary spasm was defined as total occlusion or severe vasoconstriction associated with chest pain or ischemic ST changes on the electrocardiogram or both. In Group 1, acetylcholine induced spasm in 63 (90%) of the 70 patients in the artery or arteries predicted to be responsible for spontaneous attacks. In Group 2, acetylcholine induced coronary spasm only in one patient with effort angina and advanced coronary artery disease although lesser degrees of vasoconstriction (less than or equal to 75% of the luminal diameter) occurred in most patients after acetylcholine (specificity of acetylcholine thus was 99%). In conclusion, intracoronary injection of acetylcholine is sensitive and reliable for the induction of coronary spasm. PMID- 3047197 TI - Cardiac consequences of renal transplantation: changes in left ventricular morphology and function. AB - To characterize changes in left ventricular morphology and function associated with renal transplantation, noninvasive cardiac evaluations were performed in 41 adults at the time of surgery and at follow-up. At the time of transplantation, 36 patients had undergone hemodialysis through a fistula for 2.3 +/- 2.5 years (mean +/- SD); their hematocrit level was 26 +/- 6% and systolic blood pressure was 151 +/- 19 mm Hg. Perioperatively, left ventricular hypertrophy was present in 93% of patients by echocardiography, but in only 37% by electrocardiography. Abnormal left ventricular diastolic function was present in 67% of patients and indicated a high risk for perioperative pulmonary edema. At follow-up (1.5 +/- 1.4 years), mean hematocrit level increased to 39 +/- 7%, systolic blood pressure decreased to 132 +/- 14 mm Hg and spontaneous closure of the fistula occurred in 13 patients. Left ventricular mass by echocardiography decreased from 237 +/- 66 to 182 +/- 47 g (p less than 0.001), a decrease of 23%. Left ventricular volumes and cardiac index also decreased significantly, reflecting the rapid resolution of a pretransplant high output state. Despite proportionate regression of left ventricular hypertrophy within months of transplantation, diastolic function did not improve. The significant regression of left ventricular hypertrophy that occurs after renal transplantation may help explain the improved cardiovascular survival of patients with a renal transplant over that of patients on long-term dialysis. PMID- 3047199 TI - Research methods in nutrition and dietetics: design, data analysis, and presentation. AB - Most problems in practice may be addressed through research. To show the applicability of research to all areas of nutrition and dietetics, seven types of research designs are discussed in this article: qualitative research; case series and surveys--both categorized as descriptive research; and experimental design, quasiexperimental design, cohort (follow-up) studies, and case-control studies- the four of which are categorized as analytical research because each design tests hypotheses of causal relationships. Sample size, subject selection, and statistical analysis and interpretation are discussed as appropriate to each research design. Numerous examples are presented, along with the basic research designs. Each section and subsection is numbered so that the article can serve easily as a reference and its component parts can be accessed readily. Research provides answers to questions and, generally, raises further questions that future research can address. Among the benefits of well-designed research are answers to clearly stated research questions, useful comparisons between options, information to guide evaluations of protocols, and data to document and support one's professional activities and one's staff. PMID- 3047200 TI - Food item inventory instructional simulation using microcomputers. AB - The purpose of this study was to teach the management concepts of perpetual and periodic inventory systems to university students through a food item inventory instructional simulation using microcomputers. The objective of the study was to determine the simulation's effectiveness in enhancing students' understanding of inventory systems management. The participants were senior and graduate students enrolled in a foodservice procurement and inventory systems course at Oregon State University, Corvallis. The population consisted of 68 students, with 34 in the experimental group and 34 in the control group. All the participants completed a pretest to measure existing knowledge of inventory management concepts. This was followed by two classroom lectures, each 50 minutes long, on inventory systems management in foodservice facilities. Members of the experimental group participated in using the food item inventory simulation and study guide after attending both classroom lectures. All the participants of the experimental and control groups completed a post-test 3 weeks after the pretest. The difference in post-test adjusted mean scores between the experimental and control groups was significant (p less than or equal to .05) as determined by analysis of covariance. Therefore, the food item inventory instructional simulation using microcomputers was effective in enhancing students' understanding of the management of perpetual and periodic inventory systems. PMID- 3047198 TI - Prediction of successful suppression of sustained ventricular tachyarrhythmias by serial drug testing from data derived at the initial electrophysiologic study. AB - This study investigated whether data available after the initial electrophysiologic study in patients with sustained ventricular tachyarrhythmia could identify those patients in whom serial drug testing is likely to be efficacious. One hundred six patients with inducible sustained ventricular tachyarrhythmia, whose initial study included short-term drug testing with intravenous procainamide, were evaluated. The baseline arrhythmia induced (in the absence of all antiarrhythmic drugs) was monomorphic tachycardia with a cycle length greater than 200 ms in 81 patients and ventricular flutter or fibrillation in the remaining 25 patients. After intravenous infusion of procainamide (1,250 +/- 300 mg), a ventricular tachyarrhythmia could still be induced in 80 patients during testing with up to three extrastimuli. Serial drug testing with one to four trials of oral conventional and investigational agents was then undertaken. Evaluation of 15 clinical, hemodynamic and electrophysiologic variables by stepwise logistic regression identified two independent predictors of successful response to oral antiarrhythmic drugs: 1) noninducibility of ventricular tachycardia after intravenous procainamide (p less than 0.001), and 2) left ventricular ejection fraction greater than or equal to 40% (p less than 0.05). Subgroup analysis combining each of these variables identified patients with a high, intermediate or low probability of finding a successful oral drug regimen. Patients whose arrhythmia was suppressed by intravenous procainamide had a 100% likelihood (if left ventricular ejection fraction was greater than or equal to 40%) or an 87% likelihood (if ejection fraction was less than 40%) of responding to an oral regimen.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3047201 TI - Speculations on the structure/function relationship for vagal and splanchnic afferent endings supplying the gastrointestinal tract. AB - This paper discusses some of the unsettled issues in the study of the afferent innervation of the gastrointestinal (GI) tract. Afferent fibres in the vagus and splanchnic nerves have been studied electrophysiologically and much has been learnt from single fibre recordings. Splanchnic afferent fibres generally terminate in multiple mechanosensitive endings in the mesentery and serosa where they are in a position to monitor tension on the mesenteric attachments. Other mechanoreceptors following a mainly vagal pathway behave as if they are functionally in-series with the muscle elements of the gut wall and signal muscle tension generated passively by distension and actively during contraction. A third group of afferent endings supply the GI mucosa where they are in a position to signal information on the physical and chemical environment of the gut lumen. A complex picture of mucosal sensitivity has emerged with subpopulations of receptors with polymodal sensitivity and quality-specific mechanoreceptors, thermoreceptors and chemoreceptors. Unfortunately, there is little concensus amongst the different research groups because of different experimental paradigms. One group describes specific chemoreceptors, other groups fail to find them. In this minireview I have speculated on the cause of the often conflicting data on GI afferents and the implications this has for the interpretation of visceral receptor mechanisms. PMID- 3047202 TI - [Role of macrophages and fibronectin in vitreoretinal proliferation]. AB - Proliferative vitreoretinopathy (PVR) is characterized by the periretinal proliferation of nonneoplastic cells complicated by traction retinal detachment. In this study, we report the demonstration of macrophages, which are thought to be involved in the etiology of this intraocular disease, in human periretinal membranes by an immunochemical staining procedure using a monoclonal anti-human macrophage antibody. Fibronectin, a high-molecular protein of plasma and extracellular matrix, may be visualized accordingly and could act concomitantly with macrophages as an important factor in PVR. Fibronectin is shown to be present in high quantities (Elisa) in vitreous aspirates from patients with PVR. We hypothesize that the interaction of macrophages and fibronectin may be a crucial step in the development of PVR. PMID- 3047203 TI - [Endothelial morphometry and perforating keratoplasties]. AB - 50 corneal grafts were studied by specular microscopy. The endothelial cell morphology of the grafts was analysed by using a computerised image analysis system. The endothelial cell density decreased rapidly during the first three post-operative months (17%). It continued to decrease for the first year (52%, after which cell loss occurred at a slower rate (3% of the sample cell count per year). The coefficient of variation in cell area remained constant (27%). The percentage of endothelial cell loss might be closely associated with the reestablishment of the hexagonal cellular pattern. These results suggest that the endothelium of the graft is in a state of transition during the first year after surgery. PMID- 3047204 TI - Protein modification in aging. AB - The age-related accumulation of abnormal forms of enzymes is attributable to posttranslational modification of protein structure and to a progressive loss with age of proteases that preferentially degrade the modified forms. The protein modifications include, but are not limited to: the oxidation of amino acid side chains (especially, side chains of prolyl, arginyl, lysyl and histidinyl residues) by mixed-function oxidation systems; the deamidation of asparaginyl and glutaminyl residues; the racemization and isomerization of aspartyl and asparaginyl residues; the isomerization of prolyl residues; the oxidation of cysteine sulfhydryl groups; and spontaneous changes in protein conformation that are apparently unlinked to changes in amino acid composition. Evidence supporting the roles of these protein modifications and of the proteases that degrade abnormal enzymes during aging is discussed, as well as a consideration of some technical limitations of the methods used in their study. PMID- 3047205 TI - The versatile axial pattern digital transposition flap. AB - The anatomic and clinical basis for a reliable and versatile axial pattern transposition flap for reconstruction of complex distal finger injuries is presented. This is a one-step procedure utilizing the nondominant side digital skin with possible preservation of the nerve to the fingertip. This flap obviates the necessity for a distant donor site and provides nonbulky good quality tissue. In eight patients requiring nine flaps, it has proved valuable in the salvage of severely injured digits. These injuries probably would have required distant flap reconstruction or flaps that might have jeopardized adjacent digits. PMID- 3047206 TI - Experience with Ikuta's phalangeal compression-distraction-fixation device. AB - Proper bony reduction and anatomic alignment without rotational deformity are sometimes difficult to achieve during phalangeal bone lengthening procedures. We report our experience with Ikuta's phalangeal compression-distraction-fixation device, which can be used to achieve proper bony reduction and good alignment and without any rotational deformity. PMID- 3047207 TI - Biomechanical studies of running suture for flexor tendon repair in dogs. AB - A new running-locking loop suture technique has been developed to increase tendon repair strength and to provide better tendon edge inversion. Biomechanical analysis documented the failure mechanism and the failure strength of various circumferential repair techniques. When compared with two well-known techniques, the simple circumferential running suture and Lembert running suture, the locking suture technique was shown to have 3.77 and 1.68 times greater tensile failure strength and 1.73 and 1.26 times greater stiffness than these traditional suture methods. A running peripheral locking suture may help augment the strength of tendon repair. PMID- 3047208 TI - Arthrodesis of the proximal interphalangeal joint by solid bone grafting and plate fixation in extensive injuries to the dorsal aspect of the finger. AB - Eighteen patients with industrial injuries to the dorsum of the proximal interphalangeal joint involving articular destruction and segmental bone loss were treated by primary bone grafting and plating. Rigid arthrodesis and preservation of functional length were obtained in 25 fingers. Of the 25 fingers, 23 fused primarily. Because of technical error, one reconstruction showed delayed consolidation and required secondary grafting before uniting; one arthrodesis became infected and healed by second intention. Additional procedures were flexor tendon repair, nerve grafting, and local or distant flap coverage. The procedure is considered valid for the treatment of extensive skeletal damage to the proximal interphalangeal joint area. PMID- 3047209 TI - Carpal alignment after different surgical approaches to the scaphoid: a comparative study. AB - Thirty-seven patients with inlay bone grafting for scaphoid nonunion were evaluated before and after operation for wrist function and carpal alignment. There were 26 in whom a palmar approach had been used. The remaining 11 had been treated with a dorsal approach. The two procedures showed a similar union rate (around 80%). The palmar approach, however, caused a significant increase in the scapholunate angle (p less than 0.001) and in the lunocapitate angle (p less than 0.05) and consequently augmented carpal collapse. The dorsal approach did not affect carpal alignment. The surgical division of the palmar radiocarpal ligaments, which is necessary when using a palmar approach, may be responsible for these findings. Accordingly, a dorsal approach should be preferable to a palmar approach when a graft is used for treatment of a scaphoid nonunion. PMID- 3047210 TI - Excision of the hook of the hamate: a retrospective survey and review of the literature. AB - Fractures of the hook of the hamate frequently fail to unite. In symptomatic nonunion, surgical excision of the hook has provided relief of pain in the few reported cases. In a survey of surgeons, 133 cases of excision of the hook to treat nonunion were reviewed retrospectively. The complication rate associated with excision is three percent. These complications, such as injury to the ulnar nerve or the superficial palmar arch and median nerve paresthesias secondary to retraction are due more to the surgical exposure than the actual excision. PMID- 3047211 TI - Of cholescintigraphy, sonography, and great bears. A view on modern biliary imaging. AB - We review the discrepancies and the reasons for them, in two articles in this issue of the Journal to conclude that clinical findings hold the key to selection of the proper imaging test in biliary obstruction. Cholescintigraphy is a more rewarding approach in detecting low grade obstruction, as by common duct stones, whereas in the patient with prolonged painless jaundice, and the high likelihood of a malignancy, computed tomography or ultrasound will yield the best results. Because of the potential for noninvasive imaging to miss choledocholithiasis and because of ever increasing therapeutic options, direct cholangiography will continue to be the mainstay in definition evaluation of the biliary tract. PMID- 3047212 TI - Pseudomelanosis duodeni. AB - Pseudomelanosis duodeni, speckled black pigmentation of the duodenal mucosa, presents a striking appearance at endoscopy. Among the 14 reported cases there is a predominance of black women greater than 40 years old, but it can occur in any race and age group. There is no known association with pigmentation elsewhere in the gastrointestinal tract or with the use of laxatives. However, most reported patients were hypertensive (many treated with hydralazine and propranolol) and significant numbers suffered from upper gastrointestinal bleeding, chronic renal failure, or diabetes mellitus. The pigment is usually located in mucosal macrophages, in lysosomes. Histochemical studies and electron probe microanalysis suggest that several pigments may result in this endoscopic appearance, including lipomelanin, ceroid, iron sulfide, and hemosiderin. Additional studies, possibly using tissue from surgical resections or autopsies, are needed to determine the etiology and clinical significance of this heterogeneous entity. PMID- 3047213 TI - Surgeon-related variability in the outcome of cancer surgery. AB - Statistical and clinical differences between reports of surgical therapy are usually ascribed to differences in the named method of treatment. However, it is becoming increasingly clear that surgeons vary in the ability to produce a given result; this phenomenon is sometimes referred to as "surgeon-related variability." Thus surgical treatment should be seen as the resultant vector of the named procedure plus the effect of such surgeon-related variability. These propositions have significance for the conduct of prospective randomized controlled trials which set out to test the efficacy of alternative surgical treatments. These issues are of particular importance in the cancer field because therapy is becoming more commonly multiphasic and includes both medical and surgical treatments. PMID- 3047214 TI - Influence of oral cimetidine and ranitidine on gastric emptying in active duodenal ulcer. AB - The effect on gastric emptying of 400 mg cimetidine and two doses of ranitidine (150 and 300 mg) given orally was evaluated in 45 patients with endoscopically proved duodenal ulcer and in 28 healthy controls. Gastric emptying of a radiolabeled solid phase meal was assessed. Cimetidine was confirmed not to have any significant influence on gastric emptying in duodenal ulcer patients or in healthy subjects. The significant delay in gastric emptying observed with ranitidine was dose dependent, and at the same time more pronounced in patients with duodenal ulcer than in healthy subjects. Early ulcer healing was unrelated to the changes in gastric emptying elicited by the tested drugs. PMID- 3047215 TI - Comparison of beclomethasone dipropionate and prednisolone 21-phosphate enemas in the treatment of ulcerative proctitis. AB - In a double-blind randomized clinical trial 18 patients with exacerbations of distal ulcerative colitis were treated for 4 weeks with enemas containing either prednisolone 21-phosphate 30 mg (PP) or beclomethasone dipropionate 1 mg (BDP) a surface-active corticosteroid. All 8 patients treated with PP showed clinical and endoscopic improvement in contrast with only 4 of 10 patients treated with BDP. Endocrinologic evaluation showed a significant decrease in morning plasma cortisol, in cortisol increase after synacthen, and in urinary free cortisol excretion after PP therapy, but no changes in these variables after BDP therapy. We conclude that PP enemas are more active in the treatment of ulcerative proctitis, but they cause a suppression of the adrenal cortex, in contrast to BDP. PMID- 3047216 TI - Clinical evaluation, ultrasound, cholescintigraphy, and endoscopic retrograde cholangiography in cholestasis. A prospective comparative clinical study. AB - The accuracy of clinical evaluation (CE), ultrasonography (US), and cholescintigraphy (CS) was prospectively compared in a group of 72 cholestatic patients, using the results of endoscopic retrograde cholangiography (ERC) as the true standard. Forty-six cases (63.9%) had a final diagnosis of extrahepatic biliary obstruction. Clinical evaluation had a sensitivity of 82.6% and a specificity of 46.2% for the identification of mechanical obstruction. Ultrasonography was the single most powerful noninvasive diagnostic procedure, with a sensitivity of 65.2%, a specificity of 92.3%, a positive likelihood ratio of 8.5, and a positive predictive value (PVpos) of 93.8%, which was superior to the diagnostic power of CS (0.05 less than p less than 0.1). Inconclusive results or technical failures were counted as diagnostic errors. Contingency table analyses in an extended group of 112 patients, complete for CE, US, and CS, revealed that concordant results of CE and US had a high probability of correctness (combined PVpos 92.6%, PVneg 98.0%). Contradictory ratings of these two tests were unlikely to be clarified reliably by additional CS. We conclude that the combined evaluation of CE and US allows an accurate differentiation between intrahepatic cholestasis and extrahepatic biliary obstruction. When both give contradictory results, direct cholangiography, rather than further noninvasive procedures, appears to be the diagnostic strategy of choice. PMID- 3047217 TI - Extrahepatic biliary obstruction versus intrahepatic disorder. Differentiation with hepatobiliary scintigraphy and ultrasonography. AB - In 43 patients with various hepatobiliary disorders, we compared retrospectively the sensitivity and specificity of hepatobiliary scintigraphy and ultrasonography in diagnosing extrahepatic biliary obstruction. Hepatic uptake of radioactivity from the circulation was assessed by early scintiscan at 2 min, and the clearance ratio was combined with transit time in the interpretation of hepatobiliary scintiscans. The transit time was defined as the time taken by detectable radioactivity to appear in the extrahepatic biliary tree or small intestine, whichever occurred sooner. The sensitivity and specificity were 92% and 97%, respectively. However, the specificity dropped to 74% when biliary-bowel transit time, i.e., time taken for detectable radioactivity to appear in small intestine only, was used instead of transit time in the interpretation of the scintiscans. The sensitivity and specificity of ultrasonography were 55% and 94%, respectively. We conclude that hepatobiliary scintiscan is more sensitive than ultrasonography and is reliable for diagnosing extrahepatic biliary obstruction when it is done and interpreted by the method described here. PMID- 3047219 TI - Graft-versus-host disease involves intrahepatic peribiliary glands. PMID- 3047218 TI - Sonographic demonstration of hepatic venous gas in mesenteric arterial thrombosis. AB - Hepatic portal venous gas is an ominous prognostic sign in bowel necrosis from mesenteric arterial occlusion. Ultrasonic examination is a sensitive and accurate method of demonstrating portal venous gas. We describe a case where the ultrasonic diagnosis of portal venous gas led to the rapid diagnosis of bowel infarction. Ultrasound is an appropriate emergency examination in all patients with suspected mesenteric artery occlusion. PMID- 3047221 TI - Angiodysplasia of the upper gastrointestinal tract. AB - I have reviewed the English language literature on angiodysplasia of the upper gastrointestinal tract. Angiodysplasia is a distinct mucosal vascular lesion associated with acute or chronic gastrointestinal bleeding. Its etiology is unknown, but theories of its pathogenesis have evolved from its similarity to colonic angiodysplasia and an association with renal failure. Despite recurrent gastrointestinal bleeding, the endoscopic diagnosis is difficult because the lesions are so small and so similar to fresh blood. Endoscopic ablation by electrocautery or laser photocoagulation can reduce bleeding or make it stop, but repeated treatments are often necessary. PMID- 3047220 TI - Ulcer healing after treatment with sucralfate emulsion or ranitidine. Randomized controlled study in peptic ulcer disease. AB - A randomized trial with a blind observer compared the efficacy of an emulsion containing micronized sucralfate (1 g four times daily) and ranitidine (150 mg twice daily) in the short-term healing of peptic ulcer. Patients with a minimum ulcer size of 5 mm located in the duodenal bulb, the pyloric canal, or in the prepyloric area within a distance of 2 cm from the pylorus were included. A total of 97 patients were randomized and 85 completed the trial. The endoscopically proven healing rate at 2 weeks of 41% for both sucralfate and ranitidine improved to 76% for sucralfate and 73% for ranitidine at 4 weeks and to 95% and 96%, respectively, at 12 weeks. The difference between the two treatment groups was not statistically significant, and the 95% confidence interval for the difference in ulcer healing efficacy of sucralfate emulsion compared with ranitidine was 0.15 to +0.21 at 4 weeks and -0.10 to +0.08 at 12 weeks. PMID- 3047222 TI - A problem-oriented approach to intestinal and liver disease after marrow transplantation. AB - Bone marrow transplantation has become an accepted treatment for malignancy (particularly leukemia and lymphoma), aplastic anemia, and certain inborn errors of metabolism. Patients require intensive care because of chemoradiation therapy toxicity, a prolonged period of immunosuppression and thrombocytopenia, graft versus-host disease (GVHD), and the need for parenteral nutrition. Gastrointestinal and hepatic diseases are frequent post-transplant problems. They present with intractable nausea and vomiting, intestinal bleeding, diarrhea, esophageal complaints, abdominal pain, and hepatobiliary symptoms. Our clinical approach to complex transplant patients depends on the timing of signs and symptoms after marrow grafting and on the likelihood that specific disease processes are present. Each of these major problems is covered in this review. PMID- 3047223 TI - Acute Budd-Chiari syndrome as first manifestation of adrenocortical carcinoma. AB - We report the case of a young woman in whom investigations for acute Budd-Chiari syndrome disclosed an hormone-secreting but clinically nonfunctioning adrenocortical carcinoma. We supply a very brief review of the literature. PMID- 3047224 TI - Indomethacin-associated cholestasis. AB - A patient with no prior history of liver disease developed mild cholestasis, with a moderate elevation of the serum alkaline phosphatase level, several days after instituting indomethacin therapy. The cholestasis resolved soon after the medication was discontinued. An extensive workup, including liver biopsy, revealed no alternative cause. Despite extensive use, indomethacin therapy has been associated with hepatic injury in only nine previously reported cases, as near as we can tell. PMID- 3047225 TI - A holistic technique to lower anxiety: relaxation with guided imagery. PMID- 3047226 TI - Holistic health techniques to increase individual coping and wellness. PMID- 3047227 TI - Building a body of knowledge: research on therapeutic touch, 1974-1986. PMID- 3047228 TI - Arterial chondroitin sulfate proteoglycan: localization with a monoclonal antibody. AB - We generated a monoclonal antibody (Mab) against a large chondroitin sulfate proteoglycan (CSPG) isolated from bovine aorta. This Mab (941) immunoprecipitates a CSPG synthesized by cultured monkey arterial smooth muscle cells. The immunoprecipitated CSPG is totally susceptible to chondroitinase ABC digestion and possesses a core glycoprotein of Mr approximately 400-500 KD. By use of immunofluorescence light microscopy and immunogold electron microscopy, the PG recognized by this Mab was shown to be deposited in the extracellular matrix of monkey arterial smooth muscle cell cultures in clusters which were not part of other fibrous matrix components and not associated with the cell's plasma membrane. With similar immunolocalization techniques, the CSPG antigen was found enriched in the intima and present in the medial portions of normal blood vessels, as well as in the interstitial matrix of thickened intimal lesions of atherosclerotic vessels. Immunoelectron microscopy revealed that this CSPG was confined principally to the space within the extracellular matrix not occupied by other matrix components, such as collagen and elastic fibers. These results indicate that this particular proteoglycan has a specific but restricted distribution in the extracellular matrix of arterial tissue. PMID- 3047229 TI - Demonstration of human kidney differentiation antigens with monoclonal antibodies. AB - Six human differentiation antigens (EE24.6, EG9.11, EG14.1, EI16.1, EK8.1, EK17.1) have been defined using monoclonal antibodies obtained from mice immunized with embryonic kidney cells. Their histologic distribution was determined on frozen sections of embryonic, fetal, and adult human kidneys by immunofluorescence assay. EE24.6, an ureteral bud marker, was detected only on the germ layer of mature kidney urothelium. EG9.11 and EG14.1 were detected on the S-shaped bodies and also on the adult proximal convoluted tubule for the former and the glomerular basement membrane for the latter. EI16.1, a marker of condensed mesenchyme, was detected only on epithelial cells of adult proximal convoluted tubule. EK8.1 was found in the mesangium, connective tissue, and with particularly dense labeling in the basement membranes. This labeling pattern was present throughout renal organogenesis. EK17.1 recognized both cell and plasma human fibronectins. Staining for all antibodies was nearly identical in mesonephros and metanephros. These results demonstate that some antigens follow their embryonic destiny. They indicate an antigenic similarity between the mesonephros and the metanephros and, therefore, a very early appearance of these antigens. During differentiation, these antigens concentrate on more defined structures, and staining became increased with an increased degree of differentiation. PMID- 3047232 TI - The flexible flatfoot. PMID- 3047230 TI - A novel immunohistochemical method for in vivo detection of endotoxin using horseshoe crab factor C. AB - We developed a novel immunohistochemical method for in vivo detection of endotoxin (LPS) localization in relation to the biologically active region, by use of factor C, an initiation factor in the Limulus clotting system which is mediated by LPS, as a specific affinoligand to LPS, and using rabbit anti-factor C IgG. The competitive inhibition of various LPS, lipid A, anti-LPS factor, or polymyxin B to factor C binding indicates that the immunohistochemical reaction is specific to LPS. Investigating the time course of LPS distribution during 6 hr after IV injection of 5 mg/kg to rats, the greatest uptake of LPS was evident in the reticuloendothelial system (RES), particularly in Kupffer cells, 5 min after injection, and in adrenocortical cells 3 hr after the injection. Shortly after the injection, LPS also appeared in platelet thrombi, intravascular monocytes, and a few neutrophils, and on the surface of endothelial cells in liver, kidney, spleen, lung, and aorta. Both smooth and rough forms of LPS were detectable and there was no apparent difference in their localization. This approach facilitates immunohistochemical analyses of the mechanisms involved in development of endotoxemia. PMID- 3047231 TI - Gait analysis and intoeing. PMID- 3047234 TI - Assessment of pelvic stability. PMID- 3047235 TI - Initial management of pelvic ring disruptions. PMID- 3047233 TI - Surgical management of the flatfoot. AB - The flatfoot in a child with sufficient deformity and symptoms to warrant surgery occurs infrequently. A prolonged trial of nonsurgical measures is always indicated. With the exception of the spastic paralytic foot, surgery involving alteration of the bony anatomy should be postponed until the child's foot matures. Restoration of the longitudinal arch at the midfoot naviculocuneiform articulation is a reliable solution to the symptomatic hypermobile flatfoot. The spastic paralytic foot requires tendo Achillis lengthening, medial imbrication, and Grice subtalar stabilization. The symptomatic, cartilaginous tarsal coalition will respond to resection. Treatment of the skewfoot must be individualized depending on the severity and location of symptoms, age of the patient, and joint mobility. PMID- 3047237 TI - Pelvic fractures: diagnostic and therapeutic angiography. AB - Pelvic ring disruption is part of a complex wounding pattern that challenges our ability to diagnose and manage hemorrhage. The conventional methods of diagnosis and control of abdominopelvic bleeding--peritoneal lavage and exploratory laparotomy--should be replaced by exploratory abdominopelvic angiography and transcatheter embolization. Angiography should be performed as soon as possible after the patient is admitted to the emergency room, and shock should not delay transfer of the patient to the angiography suite. PMID- 3047236 TI - Bone banks and allografts in community practice. AB - The increasing volume of orthopaedic reconstructive procedures requiring replacement of bone stock justifies the initiation of programs of bone banking in community hospitals. Provided that strict criteria are followed to assure rigorous screening of donor bone and the reliable preservation of bone graft material, community banking is safe and cost-effective. Banked allograft bone can be used successfully in a wide variety of orthopaedic procedures performed in community hospitals. In general, the best uses are filling bone cavities, buttressing, and augmenting the quantity of autograft bone. In revision reconstructive surgery of the hip, bank bone is used to replace bone stock in protrusio, acetabular dysplasia, and proximal femoral deficiency. The best and most common indication for the use of bank bone in tumor surgery is after curettage or excision of benign lesions. Allografts may be used to reconstruct bony defects after excision of malignant tumors and in the surgical treatment of metastatic disease. These instances require larger bone bank facilities than those commonly available in a community hospital setting. Medicolegal considerations related to bone banking and the use of allografts in community practice include the regulatory requirements outlined in the UAGA, questions concerning negligence liability, and theories of strict product liability. Overall, good medical practice and obtaining informed consents will minimize legal risks related to bone banking and transplantation in a community setting. PMID- 3047238 TI - Historical overview of treatment of nonunion. PMID- 3047239 TI - The use of electricity in the treatment of nonunion. PMID- 3047240 TI - Nonunion of the diaphysis of the radius and ulna. PMID- 3047241 TI - Nonunion of the humerus. AB - Under certain circumstances, fractures of the humerus may not heal. Some fractures experience delayed union and some develop nonunion despite improved methods of treatment. This chapter discussed nonunion and fracture fixation methods in the proximal, middle, and distal thirds of the humerus. Special circumstances were discussed, such as infection, nerve palsy, comminution, and electrical stimulation. PMID- 3047242 TI - Nonunion of the tibia: experience with modified Phemister bone graft and with compression plates and cancellous bone graft. PMID- 3047243 TI - Nonunions of fractures of the proximal and distal thirds of the shaft of the femur. AB - Subtrochanteric and supracondylar fractures of the femur are challenging, and specific anatomic and mechanical problems make fixation precarious. With careful selection of devices and techniques, nonunions may be avoided. Nonunion treatment requires strong immobilization and osteogenesis stimulation with bone grafts. Non invasive and semi-invasive salvage systems are not as effective as invasive procedures. The first goal of salvage is union. Joint function and muscle strength are secondary goals to be achieved later. PMID- 3047244 TI - Arthroscopy of the elbow. AB - Arthroscopy of the elbow is a technically demanding surgical procedure and attention to detail is essential. The surgeon needs a thorough knowledge of the extraarticular portal anatomy to avoid damaging nearby neurovascular structures. Several technical points need to be considered at all times, especially maintaining the elbow at 90 degrees of flexion to relax the neurovascular structures and maximally distended to move them farther away from the entering arthroscopic instruments. PMID- 3047245 TI - Arthroscopic meniscectomy. AB - Arthroscopic partial meniscectomy is indicated for unstable, irreparable tears. Meniscal repair and partial meniscectomy are not mutually exclusive concepts. These are complementary procedures, with each having specific indications. Larger, more peripheral tears are repaired, whereas smaller, more central irregular or irreparable tears are treated by arthroscopic partial meniscectomy. The emphasis is on leaving a well-contoured stable meniscus while excising only the unstable or damaged portion. The remaining meniscus may still provide some protection for the hyaline cartilage of that compartment. PMID- 3047247 TI - Problem fractures and dislocations of the hand. PMID- 3047246 TI - Arthroscopic meniscus repair with a posterior incision. AB - The transarticular arthroscopic approach with a posterior incision provided a method of repairing more than 98% of unstable meniscus tears encountered between November 1983 and November 1986. A clinically stable bond was obtained in most of these tears with a subjective failure rate of 2% or less. There was a trend towards better healing of isolated meniscus repairs and lateral meniscus tears less than eight weeks old associated with ACL reconstruction when a blood clot injection was used to supplement the rasp abrasion of the parameniscal synovium. Healing of rim widths to 5 mm can be obtained with these methods. Indications for meniscus repair include all lateral meniscus tears and all medial meniscus tears except when repair of a stump would not replace 25% or more of the missing area. In our experience, this includes more than 98% of all unstable meniscus tears. Contraindications to meniscus repair include short (10 mm or less) stable tears, partial thickness (less than 50% of vertical height), and shallow radial tears (3 mm or less in depth). The posterior incision and popliteal retractor are necessary to protect the popliteal neurovascular structures. PMID- 3047248 TI - Classification of ankle fractures: which system to use? PMID- 3047249 TI - Management of open fractures in the multiply injured patient. PMID- 3047250 TI - Blood supply to bone and proximal femur: a synopsis. PMID- 3047251 TI - Injuries to the muscle-tendon unit. AB - Injuries to muscles are very common in clinical practice. Our understanding of the pathophysiology lags far behind that of other orthopaedic injuries. The recent increase in research and emphasis on muscle injuries and sports medicine will, it is hoped, provide new data and a more scientific basis for treatment and prevention. PMID- 3047252 TI - Handling malpractice stress: suggestions for physicians and their families. PMID- 3047253 TI - A system of staging musculoskeletal neoplasms. PMID- 3047254 TI - The staging of aseptic necrosis. PMID- 3047255 TI - Management of avascular necrosis of the femoral head--an overview. AB - The most common cause of AVN of the femoral head is a displaced subcapital or transcervical fracture. Circumstances here differ significantly from those encountered in the patient with nontraumatic AVN, especially because the traumatic variety is seen most often in the elderly population. Attempts to preserve the femoral head are generally not indicated and the treatment of choice for the hip with sufficient pain and disability is either endoprosthetic replacement or total hip replacement. In contradistinction, nontraumatic AVN occurs primarily in younger adults and is often bilateral. In these patients the goal is to preserve, not to replace, the femoral head. "Conservative" or nonsurgical management has generally been unsuccessful. Results with established surgical procedures have been inconsistent and frequently disappointing, often because they have been instituted too late. Although no approach is completely effective, in general the earlier treatment begins, the better the results will be. Early diagnosis, therefore, is essential. This depends on a heightened clinical awareness of this condition. The importance of various imaging techniques, including high-quality plain radiographs, technetium scans, and magnetic resonance imaging must be stressed. Newer procedures for the treatment of early AVN are being developed and evaluated. These currently include the use of vascularized grafts, osteochondral allografts, transtrochanteric rotational osteotomy, and electrical stimulation. I hope that one or more of these approaches will prove useful in preserving the femoral head. Until such time, I shall continue to use bone grafting with decompression to treat hips with limited involvement. When definite collapse of the femoral head has occurred and when pain and disability are sufficient to require surgical intervention, the treatment of choice is arthroplasty. Although many still favor the use of a bipolar femoral endoprosthesis, I believe that total hip replacement provides more consistent, more durable, and better results. Whichever arthroplasty is chosen, I anticipate continually improving results with advances in technique and component design. PMID- 3047256 TI - Early diagnosis of avascular necrosis of the femoral head. AB - Early diagnosis is important because it leads to early treatment and a better chance of saving the femoral head. The first, and perhaps the most important step, in making an early diagnosis is a heightened clinical awareness of this condition. The physician must be familiar with the nonspecific symptoms, physical findings, and laboratory tests, as well as with the various predisposing factors associated with avascular necrosis. Other causes of hip abnormality or symptoms must be ruled out, as AVN is in part a diagnosis of exclusion. Imaging techniques, which are extremely useful in the early diagnosis of this condition, include good-quality plain radiographs, technetium bone scans, and especially MRI. Technetium bone scans and MRI are quite sensitive, although not entirely specific, and must be used in conjunction with a comprehensive clinical evaluation of the patient. PMID- 3047257 TI - Avascular necrosis of the femoral head in children. AB - Two types of AVN occur in childhood. The first is the idiopathic type best characterized by Legg-Calve-Perthes' disease associated with repeated infarctions that slow the healing process. The second is the traumatic type caused by a direct insult to the blood supply of the developing femoral head. Both can result in profound growth disturbances of the proximal femur and early degenerative changes. They may have little effect on hip function or appearance. The variables affecting these outcomes are the age of the child, the degree of femoral head involvement, the presence of at-risk signs, the ability of the acetabulum to contain the healing head, and whether or not the growth plate closes as a result of the AVN. Bone scans and magnetic resonance imaging offer some potential for improved accuracy in diagnosis and prediction. The recommendation treatment for AVN in children is to treat the synovitis whenever it occurs or recurs. Abduction bracing should be used for the first six months in most young children with idiopathic AVN of the Legg-Calve-Perthes' type. Protected weightbearing with crutches should suffice for the early phases of traumatic AVN. Within six months, one should be able to determine the classification and whether to continue the treatment being used or to change to another form of treatment. Petrie casts are used rarely but can be very useful in certain patients. Prolonged crutch use may help to prevent collapse while relieving symptoms in the older child.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3047259 TI - Surgical correction of clubfoot. PMID- 3047258 TI - The painful foot in the child. PMID- 3047260 TI - Pathoanatomy and surgical treatment of the resistant clubfoot. PMID- 3047261 TI - Amperometric enzyme-amplified immunoassays. AB - Enzyme-amplified immunoassays have been adapted for electrochemical measurement, using an NAD+/NADH redox cycle coupled to an electrode via the active site of diaphorase. Two amperometric methods are described, the first employs an organic conducting salt electrode, NMP+/TCNQ-; the second a platinum wire with ferricyanide as electron transfer mediator. In an immunoenzymometric assay for human prostatic acid phosphatase the sensitivities of the electrochemical methods were comparable to that achieved with the existing optical technique, but the dynamic range of the electrochemical assays was increased by at least two orders of magnitude. It is proposed that electrochemical enzyme-amplified immunoassays may eventually replace their optical counterparts. PMID- 3047262 TI - Malaria sporozoite penetration. A new approach by double staining. AB - To determine, whether a sporozoite is outside the hepatocyte membrane or internalized, a double staining test was carried out using, successively, antibody labeled with peroxidase and fluorescein. This test permits the quantification of sporozoite entry and outline sporozoite-hepatocyte interactions. PMID- 3047263 TI - Improvements in the purification of an antisteroid antiserum resistant to conventional immunoadsorption chromatography. AB - Antibodies against dexamethasone, a synthetic steroid, have been induced in rabbits immunized with a 3-carboxymethyloxime dexamethasone derivative conjugated to bovine serum albumin. The antiserum displaying the highest affinity for dexamethasone (KD = 0.5 nM) appeared to be resistant to purification on an agarose matrix bearing the same 3-carboxy-methyloxime dexamethasone derivative. No desorption of active antibodies could be obtained whatever the eluting buffer used. Electrophoretic elution gave only poor results. However a very significant improvement in the purification of these antibodies was achieved by changing the connecting arm for steroid linkage to the agarose beads. A 17-fold purification and a 32% recovery of active specific antisteroid antibodies were obtained using a column bearing a 17 beta-carboxamide dexamethasone derivative. Moreover good results (23-fold purification and 30% recovery) were also obtained with a commercially available preactivated high-performance silica column derivatized with an aminated 3-carboxymethyloxime derivative of dexamethasone. In this case the more efficient diffusion of the eluting solution through the pores of a high performance stationary phase made of small diameter rigid beads probably explained the strikingly improved results, when compared to those obtained with agarose beads bearing the same dexamethasone derivative. PMID- 3047264 TI - Indirect immunofluorescence labelling with complexes of phycoerythrin and monoclonal anti-phycoerythrin antibodies (PEAPE complexes). AB - We describe a method of immunofluorescence which is a lateral application of the principles of the APAAP immunohistochemical technique. Immune complexes of R phycoerythrin and monoclonal anti-R-phycoerythrin (PEAPE complexes) were used in an indirect immunofluorescence technique to detect the binding to cells of monoclonal antibodies directed to IgM, HLA-DR and B cell activation and differentiation antigens. PEAPE complexes were linked to cell surface bound mAbs by unlabelled anti-mouse Ig antibodies to produce high levels of fluorescent staining. The sensitivity of this method of indirect immunofluorescence was enhanced by the sequential application of several cycles of anti-mouse Ig and PEAPE complexes. PMID- 3047265 TI - Rapid detection of low levels of donor specific IgG by flow cytometry with single and dual colour fluorescence in renal transplantation. AB - Lymphocytotoxic immunoglobulin is routinely assayed before human renal transplantation. If IgG directed against donor T cells is detected in the serum of the potential recipient, transplantation is not performed as it is associated with a poor graft outcome. Poor sensitivity of the conventional assay has been postulated as being the cause of some graft failures. Two new flow cytometric assays are described which are more sensitive than the conventional test. The first assay requires manual separation of T and B lymphocytes and therefore takes a similar time to perform as the conventional assay. The second assay utilises a two-colour system and lymphocyte's separation is by fluorescence. This assay takes half the time to perform, thereby decreasing graft ischaemic time before transplantation. PMID- 3047266 TI - Hexamethylene bisacetamide-induced differentiation of transformed cells: molecular and cellular effects and therapeutic application. AB - Hexamethylene bisacetamide (HMBA), a highly polar compound, induces murine erythroleukemia (MEL) cells to express the erythroid phenotype, including cessation of proliferation. Inducer-mediated differentiation of MEL (DS19) cells is a multistep process characterized by a latent period during which a number of changes occur including alterations in ion flux, an increase in membrane-bound protein kinase C (PKC) activity, the appearance of Ca2+ and phospholipid independent PKC activity in the cytosol, and modulation in expression of a number of genes such as c-myc, c-myb, c-fos and the p53 genes. HMBA-mediated commitment to terminal differentiation is first detected at about 12 hours and increases in a stochastic fashion until over 95% of the population is recruited to terminal differentiation by 48 to 60 hours. Commitment is associated with persistent suppression of c-myb gene expression. By 36 to 48 hours, transcription of the globin genes has increased 10 to 30 fold, whereas transcription from rRNA genes is suppressed. The steroid, dexamethasone, or the tumor promoter, phorbol-12 myristate-13-acetate (TPA), suppress HMBA-induced MEL cell terminal differentiation. These agents appear to act at a late step during the latent period. MEL cell lines derived from DS19 by selection for resistance to vincristine are: 1) induced to commit without a detectable latent period, 2) markedly more sensitive to HMBA, and 3) resistant to dexamethasone or TPA inhibition of HMBA-induced commitment. The data suggests that vincristine resistant MEL cells express a factor which circumvents essential HMBA-mediated early events. In vitro studies with HMBA provide a basis for the application of HMBA to clinical therapy of human cancers. Clinical trials with HMBA have been initiated. PMID- 3047268 TI - Current prevalence of trichinosis in pigs in Egypt. PMID- 3047267 TI - Thymic dependency of the humoral regulation of CFU-s proliferation in mice bearing a CSF-producing tumor. AB - A better understanding of the mechanisms involved in the proliferation of splenic colony-forming units (CFU-s) during tumor growth is important for the prevention of bone marrow aplasia during chemotherapy. The in vivo growth of EMT6 cells, a colony-stimulating factor-secreting mammary tumor, in BALB/c and nude mice resulted in splenomegaly and an increase in the number of splenic granulocyte/macrophage colony-forming cells (GM-CFC). Proliferation of CFU-s, observed in BALB/c mice but not in nude mice, most likely resulted from combined direct and indirect actions of factors secreted by tumor and host cells (in particular helper T cells). These factors were detectable in the serum immediately following tumor cell injection. Thus, the GM-CFC response to factors secreted by the EMT6 tumors is thymus-independent while the CFU-s response is dependent upon the presence of T cells. Finally, we show that EMT6 tumor growth had no effect on the determination of CFU-s differentiation toward the various myeloid cell lineages. PMID- 3047269 TI - Toxoplasma infection of cats in Cairo area as revealed by IFAT. PMID- 3047271 TI - Evidence of a biological mode of transmission of enteric bacteria by Musca domestica sorbens. PMID- 3047270 TI - Experimental contamination of Musca domestica sorbens in relation to external and gut pathogen transmission. PMID- 3047272 TI - Seroepidemiologic observations on human toxoplasmosis in Dakahlia Governorate. PMID- 3047273 TI - Diagnosis of bancroftian filariasis by detection of circulating antigens by counterimmunoelectrophoresis. PMID- 3047274 TI - Humoral and cellular responses in experimental trichinosis. PMID- 3047275 TI - A review and distribution map of rodents in Sinai, Egypt. PMID- 3047276 TI - Nutrition in cancer patients: an update and review of our experience. Issues in symptom control. Part 3. PMID- 3047277 TI - [Hemodynamic evaluation of long-term follow-up prosthetic valves in mitral position using Doppler ultrasound and echocardiography--with special reference to types and sizes]. PMID- 3047278 TI - [The experimental studies on the myocardial preservation during the reperfusion period--the recovery of the myocardial temperature and the effectiveness of coronary reperfusion using the glucose-insulin solution]. PMID- 3047279 TI - [A case of combined esophago-bronchopulmonary cyst]. PMID- 3047280 TI - [Successful surgical treatment in a 79-year-old woman with post myocardial infarction septal perforation]. PMID- 3047281 TI - [Immunohistochemical study on the proliferative potential of trophoblast in early pregnancy using the monoclonal antibody Ki-67]. PMID- 3047282 TI - Infectivity of secondary dapsone-resistant cases. AB - The incidence rate of leprosy among 517 household contacts of 113 cases of secondary dapsone resistance with 5074 person years at risk were studied. The incidence rate of leprosy was 4.3 per 1000 person years at risk, which is very similar to the incidence rate (4.8) among household contacts of lepromatous cases. Two, possibly three, cases of primary dapsone resistance were detected among the 27 contacts who developed multibacillary leprosy. There was no evidence of dapsone resistance among 48 paucibacillary leprosy cases assessed when treated with dapsone monotherapy. The possibility that secondary dapsone-resistant cases will infect and will result in an increase in the number of primary dapsone resistant cases needs to be investigated further. PMID- 3047284 TI - Transitory macrophage activation in the granulomatous lesions of Mycobacterium lepraemurium-induced lepromatoid leprosy in the mouse. AB - A kinetic study on the evolution of granulomas that appear in the liver of NIH mice inoculated with 10(8) Mycobacterium lepraemurium by the intraperitoneal route has been performed. The liver was chosen because of its nonlymphoid histology which allowed us to visualize the appearance and maturation of the cell infiltrates generated as a consequence of the mycobacterial infection. The study analyzed both the macrophage activation within the granulomas and the fate of bacilli within the macrophage. The results showed that this mycobacteriosis induces a relatively early macrophage activation (a very likely result of a cell mediated immune response triggered by the bacilli) that peaks between 45 and 60 days postinoculation, fades thereafter, and practically disappears several days later. Bacilli are susceptible to the microbicidal effects of activated macrophages, but when the macrophages are turned off (probably due to active suppressive mechanisms), the surviving bacilli reinitiate the infection with no further macrophage opposition. As a result, more phagocytes are attracted to the infection sites and the cell infiltrates grow steadily to become confluent, increasing the granuloma fraction and eventually replacing the liver parenchyma. The findings suggest that in murine "leprosy" infection, early immunological changes occur that enable the macrophages present in the granulomas to kill the infecting M. lepraemurium regardless of the eventual lepromatoid evolution of the granulomas. Lepromatoid granulomas in the mouse and lepromatous granulomas in man are equivalent structures in regard to their histology and bacteriology. PMID- 3047283 TI - Armadillo IgG and IgM antibody responses to phenolic glycolipid-I during experimental infection with M. leprae. AB - The kinetics of antibody responses of Mycobacterium leprae-infected armadillos to phenolic glycolipid-I (PGL-I) were studied by means of ELISA. The levels of both IgG and IgM antibodies to PGL-I increased with time. Some animals were less susceptible to disseminations of M. leprae infection and lived longer than others. These animals had high absorbance values (greater than 0.7) for IgG anti PGL-I compared to more susceptible armadillos that had lower absorbance values for IgG anti-PGL-I. PMID- 3047286 TI - Immunological approach for control of Mycobacterium avium-intracellulare infections in AIDS-an hypothesis. PMID- 3047285 TI - Histopathological changes in the eyes of mangabey monkeys with lepromatous leprosy. AB - Leprosy is the third leading cause of preventable blindness; however, little is known about the spread of infection to the eye. We have studied the eyes of three sooty managabey monkeys. Two were experimentally infected with Mycobacterium leprae; the third was not infected. In one of the infected animals there was histopathological evidence of lepromatous leprosy as evidenced by a chronic inflammatory infiltrate at the limbus, and detection of acid-fast bacilli in the corneal stroma, blood vessel walls, and corneal nerves. The latter were damaged as a result of the bacillary invasion. Electron microscopy revealed involvement and distortion of keratocytes with M. leprae and invasion of the corneal stroma by macrophages containing bacilli. Both infected animals showed focal collections of lymphocytes in the superficial stroma of the conjunctiva and in the ciliary body. This is the first report of the ocular manifestations of leprosy in any primate, including man, in which the duration of infection is known. PMID- 3047287 TI - The activity against yeasts of human cerumen. PMID- 3047288 TI - Chlamydia: its influence in chronic secretory otitis media. AB - Since Chlamydia trachomatis was isolated from middle ear effusions of neonates with natally acquired chlamydial infection (Tipple et al., 1979), there have been several studies to detect chlamydia in older children with chronic secretory otitis media, mainly by tissue culture. In this study, the aspirates of 106 middle ear effusions of 60 children with chronic secretory otitis media were investigated for the presence of C. trachomatis, other bacteria and viruses. An amplified enzyme-linked immunoassay was used to detect the presence of chlamydia. The bacteriological and virological results mirrored previous studies in the United Kingdom and no chlamydia were found. Chlamydia do not appear to be related to the aetiology of this disease in the population examined. PMID- 3047289 TI - The Indian contribution to rhinoplasty. PMID- 3047290 TI - Chronic Streptococcus pyogenes tonsillitis, with rheumatic arthritis and myocarditis (an immunological study using the SWT). PMID- 3047291 TI - Surgical salvage for squamous cancer involving the supraglottic larynx. AB - Anatomically the supraglottic larynx has been shown to be self contained as regards its boundaries and lymphatic compartments which tend to limit the spread of cancer arising within the region until it reaches the margins of the supraglottis (Pressman and Simon, 1961). These may be called Type A tumours. Advanced glottic tumours secondarily involving the supraglottis may be considered a sub-group within the larynx (transglottic). PMID- 3047292 TI - Haemangioma of the maxilla. AB - Haemangioma involving the paranasal air sinuses is rare. It presents with severe epistaxis or bleeding due to dental extraction and may mimic malignancy. A case of haemangioma of the maxilla is presented and the diagnostic and operative difficulties together with a review of literature are discussed. Bucy and Capp (1930) described primary haemangioma of bone with special reference to X-ray diagnosis. Wyke (1949) was of the opinion that haemangiomas accounted for only 10 per cent of primary benign neoplasms of skull bones. Batsakis (1979) commented that haemangioma of bone accounted for seven per cent of all osseous neoplasms and that from a review of literature he had found no more than 40 such cases. Dahlin (1967) in a review of 3,947 cases of bone tumours, found 47 cases of osseous haemangioma out of which only three involved the upper jaw. Smith (1959) reported that only 10 cases of haemangioma of the maxilla had been published by 1959 and further isolated cases have been described by Sawhney et al. (1973), Pandhi et al. (1977) and Ahad and Chisti (1977). PMID- 3047293 TI - Chondrosarcoma of the sphenoid--a case report and review. AB - The case of 57-year-old woman is presented with a chondrosarcoma of the sphenoid. The main symptom was rapid loss of vision and in order to halt this the tumour was debulked. The literature relating to chondrosarcomas of the head and neck is reviewed. These rare tumours are not radio-sensitive and surgery represents the best form of treatment at the present time. The prognosis is poor. PMID- 3047294 TI - Acquisition of domain-specific knowledge in patients with organic memory disorders. PMID- 3047295 TI - A research review and alternative hypothesis explaining the link between learning disability and delinquency. PMID- 3047296 TI - Evaluating the field test revision process by comparing two versions of a reasoning skills CAI program. PMID- 3047297 TI - Standards for animal research: looking at the middle. AB - Much of the public debate over laboratory animal use has focused on either the scientist's demand for absolute freedom of inquiry, or the abolitionist's demand for an end to animal use in science. Yet many recent proposals for reform seek instead to balance the interests of laboratory animals in avoiding harm against the interests of research beneficiaries in continued animal use. This essay is an analysis of the intermediate reform positions and their underlying ethical principles. PMID- 3047299 TI - Alterations in the viscosity and deformability of red cells in patients with Plasmodium falciparum. PMID- 3047300 TI - Fetal sex determination by ultrasonography. PMID- 3047301 TI - Colonic endometriosis: a case report. PMID- 3047298 TI - Comparison of the results of the neurorrhaphic techniques; epineurial, fascicular and combined epineurial-fascicular neurorrhaphy. PMID- 3047302 TI - Prenatal diagnosis: a modern technology for fetal diagnosis, prevention and treatment. PMID- 3047304 TI - Clinical trials of intramuscular sulprostone for second trimester abortion. PMID- 3047303 TI - A comparative study of tramadol and pethidine in laparoscopic interval sterilization. PMID- 3047305 TI - Monoclonal antibody pregnancy test. PMID- 3047306 TI - Fever in children younger than three months of age. A pooled analysis. AB - Concern that febrile infants younger than 3 months of age are at high risk of serious infection has prompted a management policy of routine hospitalization with antibiotic administration. Ten published studies of febrile infants younger than 3 months of age were reviewed, and data were statistically combined to develop estimates of the risk of bacteremia and serious infection. Factors that predicted increased risk were similarly evaluated. Mean and median risk estimates included, respectively, 3.0 and 3.4 percent for bacteremia, 1.3 and 1.0 percent for septic meningitis, and 5.0 and 7.0 percent for pneumonia. These were no higher than comparable estimates for older infants. Clinical appearance was 92 percent sensitive in predicting bacteremia in 500 infants (23 of 25 cases). Younger age, higher fever, and elevated white blood cell count were associated with increased risk of serious infection. Data from these studies do not support the belief that febrile infants younger than 3 months are uniformly at greater risk of serious infection than older infants. Judicious evaluation of younger infants could lead to more selective, cost-efficient management. PMID- 3047307 TI - Is screening mammography routinely indicated for women between 40 and 50 years of age? An opposing view. PMID- 3047308 TI - Renin release by afferent arterioles incubated in vitro. PMID- 3047309 TI - Absorption and effectiveness of mixtures of short- and intermediate-acting insulins. PMID- 3047310 TI - Traumatic pseudocysts of the pancreas in children: report of 3 cases. PMID- 3047312 TI - Interactions of bromide, iodide, and fluoride with the pathways of chloride transport and diffusion in human neutrophils. AB - Isolated human neutrophils possess three distinct pathways by which Cl- crosses the plasma membrane of steady state cells: anion exchange, active transport, and electrodiffusion. The purpose of the present work was to investigate the selectivity of each of these separate processes with respect to other external halide ions. (a) The bulk of total anion movements represents transport through an electrically silent anion-exchange mechanism that is insensitive to disulfonic stilbenes, but which can be competitively inhibited by alpha-cyano-4 hydroxycinnamate (CHC; Ki approximately 0.3 mM). The affinity of the external translocation site of the carrier for each of the different anions was determined (i) from substrate competition between Cl- and either Br-, F-, or I-, (ii) from trans stimulation of 36Cl- efflux as a function of the external concentrations of these anions, (iii) from changes in the apparent Ki for CHC depending on the nature of the replacement anion in the bathing medium, and (iv) from activation of 82Br- and 125I- influxes by their respective ions. Each was bound and transported at roughly similar rates (Vmax values all 1.0-1.4 meq/liter cell water.min); the order of decreasing affinities is Cl- greater than Br- greater than F- greater than I- (true Km values of 5, 9, 23, and 44 mM, respectively). These anions undergo 1:1 countertransport for internal Cl-. (b) There is a minor component of total Cl- influx that constitutes an active inward transport system for the intracellular accumulation of Cl- [( Cl-]i approximately 80 meq/liter cell water), fourfold higher than expected for passive distribution. This uptake is sensitive to intracellular ATP depletion by 2-deoxy-D-glucose and can be inhibited by furosemide, ethacrynic acid, and CHC, which also blocks anion exchange. This active Cl- uptake process binds and transports other members of the halide series in the sequence Cl- greater than Br- greater than I- greater than F- (Km values of 5, 8, 15, and 41 mM, respectively). (c) Electrodiffusive fluxes are small. CHC-resistant 82Br- and 125I- influxes behave as passive leak fluxes through low-conductance ion channels: they are nonsaturable and strongly voltage dependent. These anions permeate the putative Cl- channel in the sequence I- greater than Br- greater than Cl- with relative permeability ratios of 2.2:1.4:1, respectively, where PCl approximately 5 X 10(-9) cm/s. PMID- 3047311 TI - Characterization of cross-bridge elasticity and kinetics of cross-bridge cycling during force development in single smooth muscle cells. AB - Force development in smooth muscle, as in skeletal muscle, is believed to reflect recruitment of force-generating myosin cross-bridges. However, little is known about the events underlying cross-bridge recruitment as the muscle cell approaches peak isometric force and then enters a period of tension maintenance. In the present studies on single smooth muscle cells isolated from the toad (Bufo marinus) stomach muscularis, active muscle stiffness, calculated from the force response to small sinusoidal length changes (0.5% cell length, 250 Hz), was utilized to estimate the relative number of attached cross-bridges. By comparing stiffness during initial force development to stiffness during force redevelopment immediately after a quick release imposed at peak force, we propose that the instantaneous active stiffness of the cell reflects both a linearly elastic cross-bridge element having 1.5 times the compliance of the cross-bridge in frog skeletal muscle and a series elastic component having an exponential length-force relationship. At the onset of force development, the ratio of stiffness to force was 2.5 times greater than at peak isometric force. These data suggest that, upon activation, cross-bridges attach in at least two states (i.e., low-force-producing and high-force-producing) and redistribute to a steady state distribution at peak isometric force. The possibility that the cross-bridge cycling rate was modulated with time was also investigated by analyzing the time course of tension recovery to small, rapid step length changes (0.5% cell length in 2.5 ms) imposed during initial force development, at peak force, and after 15 s of tension maintenance. The rate of tension recovery slowed continuously throughout force development following activation and slowed further as force was maintained. Our results suggest that the kinetics of force production in smooth muscle may involve a redistribution of cross-bridge populations between two attached states and that the average cycling rate of these cross-bridges becomes slower with time during contraction. PMID- 3047314 TI - Anticonvulsant drug actions on neurons in cell culture. AB - Two actions of clinically used antiepileptic drugs have been studied using mouse neurons in primary dissociated cell culture. The antiepileptic drugs phenytoin, carbamazepine and valproic acid were demonstrated to limit sustained high frequency repetitive firing of action potentials at free serum concentrations that are achieved in patients being treated for epilepsy. Furthermore, an active metabolite of carbamazepine also limited sustained high frequency repetitive firing while inactive metabolites of phenytoin and carbamazepine did not limit sustained high frequency repetitive firing. Phenobarbital and clinically used benzodiazepines limited sustained high frequency repetitive firing of action potentials, but only at concentrations achieved during the treatment of generalized tonic-clonic status epilepticus. Ethosuximide did not limit sustained high frequency repetitive firing even at concentrations four times those achieved in the serum of patients treated for generalized absence seizures. Phenobarbital and clinically used benzodiazepines enhanced postsynaptic GABA responses at concentrations achieved free in the serum during treatment of generalized tonic clonic or generalized absence seizures. However, phenytoin, carbamazepine, valproic acid and ethosuximide did not modify postsynaptic GABA responses at therapeutic free serum concentrations. These results suggest that the ability of antiepileptic drugs to block generalized tonic-clonic seizures and generalized tonic-clonic status epilepticus may be related to their ability to block high frequency repetitive firing of neurons. The mechanism underlying blockade of myoclonic seizures may be related to the ability of antiepileptic drugs to enhance GABAergic synaptic transmission. The mechanism underlying management of generalized absence seizures remains unclear. PMID- 3047313 TI - The organization of taste sensibilities in hamster chorda tympani nerve fibers. AB - Electrophysiological measurements of nerve impulse frequencies were used to explore the organization of taste sensibilities in single fibers of the hamster chorda tympani nerve. Moderately intense taste solutions that are either very similar or easily discriminated were applied to the anterior lingual surface. 40 response profiles or 13 stimulus activation patterns were considered variables and examined with multivariate statistical techniques. Three kinds of response profiles were seen in fibers that varied in their overall sensitivity to taste solutions. One profile (S) showed selectivity for sweeteners, a second (N) showed selectivity for sodium salts, and a third (H) showed sensitivity to salts, acids, and other compounds. Hierarchical cluster analysis indicated that profiles fell into discrete classes. Responses to many pairs of effective stimuli were covariant across profiles within a class, but some acidic stimuli had more idiosyncratic effects. Factor analysis of profiles identified two common factors, accounting for 77% of the variance. A unipolar factor was identified with the N profile, and a bipolar factor was identified with the S profile and its opposite, the H profile. Three stimulus activation patterns were elicited by taste solutions that varied in intensity of effect. Hierarchical cluster analysis indicated that the patterns fell into discrete classes. Factor analysis of patterns identified three common unipolar factors accounting for 82% of the variance. Eight stimuli (MgSO4, NH4Cl, KCl, citric acid, acetic acid, urea, quinine HCl, HCl) selectively activated fibers with H profiles, three stimuli (fructose, Na saccharin, sucrose) selectively activated fibers with S profiles, and two stimuli (NaNO3, NaCl) activated fibers with N profiles more strongly than fibers with H profiles. Stimuli that evoke different patterns taste distinct to hamsters. Stimuli that evoke the same pattern taste more similar. It was concluded that the hundreds of peripheral taste neurons that innervate the anterior tongue play one of three functional roles, providing information about one of three features that are shared by different chemical solutions. PMID- 3047315 TI - Anti-epileptic effects of focal micro-injection of excitatory amino acid antagonists. AB - The role of excitatory synaptic activity at various brain regions in the development and spread of seizure activity has been investigated by the focal microinjection of 2-amino-7-phosphono-heptanoate (2-APH), a selective antagonist at the N-methyl-D-aspartate preferring receptor, or gamma-D-glutamyl-aminomethyl sulphonate (GAMS), a partially selective antagonist at the kainate receptor. In genetically epilepsy prone rats the seizure response to a loud sound in most effectively suppressed by focal injections of 2-APH, 0.1-1.0 nmol, in the inferior colliculus. Protection is also seen after injections of 2-APH, 25 nmoles, in the substantia nigra (pars reticulata) or the midbrain reticular formation. Motor limbic seizures induced by pilocarpine, 380 mg/kg intraperitoneally, are prevented by prior injection into the substantia nigra, pars reticulata, or the entopeduncular nucleus, of 2-APH, 10 nmol or 10 pmol, respectively. Similar protection follows the injection of 2-APH, 1-5 pmol in the piriform cortex. The convulsant effects of pilocarpine are also blocked by the focal injection of GAMS, 10 nmol in the entopeduncular nucleus. This experimental approach can indicate critical sites at which seizure activity is initiated in particular models (e.g., inferior colliculus in sound-induced seizures, and piriform cortex in limbic seizures) and the pathways controlling seizure expression, such as the basal ganglia outputs. It also identifies specific receptors at which anticonvulsant drugs may operate. PMID- 3047316 TI - Immunohistochemical localization of Ca2+/calmodulin-dependent protein kinase II in rat brain and various tissues. AB - Polyclonal antibodies against Ca2+/calmodulin-dependent protein kinase II (CaM kinase II) of rat brain were prepared by immunizing rabbits and then purified by antigen-affinity column. The antibodies which recognized both subunits of the enzyme with Mrs 49K and 60K were used for the study on the distribution of CaM kinase II in formalin-fixed, paraffin-embedded tissues. In the brain, a light microscopic study demonstrated strong immunoreactivity in neuronal somata and dendrites and weak immunoreactivity in nuclei. The densely stained regions included cerebral cortex, hippocampal formation, striatum, substantia nigra, and cerebellar cortex. In substantia nigra, neurites were stained, but not neuronal somata. Electron microscopy revealed that the immunoreactive product was highly concentrated at the postsynaptic densities. In addition to neurons, weak immunoreactivity was also demonstrated in glial cells, such as astrocytes and ependymal cells of ventricles and epithelial cells of choroid plexus. In other tissues, strong immunoreactivity was observed in the islet of pancreas and moderate immunoreactivity in skeletal muscle and kidney tubules. Immunoreactivity was demonstrated in all of the tissues tested. The results suggest that CaM kinase II is widely distributed in the tissues. PMID- 3047317 TI - Glial fibrillary acidic protein increases in the spinal cord of Lewis rats with acute experimental autoimmune encephalomyelitis. AB - Glial fibrillary acidic protein (GFAP) in the spinal cords of Lewis rats with acute experimental autoimmune encephalomyelitis (EAE) was quantitated by densitometry of both stained gels and immunoblots of electrophoretically separated cytoskeletal proteins. The experimental period ranged from 7 to 65 days postinoculation (dpi). Greater than 92% of the total spinal cord GFAP was recovered in the Triton-insoluble cytoskeletal pellet; less than 2% was truly soluble. GFAP increased gradually and significantly with time, reaching a level one-and-a-half to two times greater than that of controls by 35 dpi and remaining elevated at 65 dpi. In EAE animals, GFAP was 33% of the total Triton-insoluble protein (excluding histones and other small basic proteins) at 7 dpi, rising to 48% at 65 dpi. Increases in vimentin were also noted, following a time course similar to that of GFAP. An increase in immunocytochemical staining of GFAP was noticeable at 10 dpi and became marked at 14 dpi, a time before GFAP levels had increased significantly. Thus, enhanced staining at the peak of the disease cannot be explained simply by an increase in antigen protein. Other possible explanations, such as an increase in soluble GFAP content, proteolytic degradation, or modifications in the immunochemical properties of GFAP in EAE animals, were ruled out. Both the biochemical and immunocytochemical increases in GFAP persisted through 65 dpi, even though the animals recovered from clinical signs at approximately 18 dpi. PMID- 3047318 TI - Purifications of rat adrenal dopamine-beta-hydroxylase: immunological analysis of its soluble and membrane-bound forms with the use of an antibody raised against the soluble form. AB - Soluble and membrane-bound dopamine-beta-hydroxylases (sDBH and mDBH, respectively) from rat adrenal glands have been purified through concanavalin A Sepharose chromatography, gel filtration, and ion-exchange high-performance chromatographies. Both sDBH and mDBH were composed by four subunits of apparent molecular weight of 75,000 and showed a native molecular weight of 300,000. This procedure has not allowed us to obtain a sufficient amount of enzyme to immunize a rabbit. A second, more rapid procedure was designed to isolate sDBH, including concanavalin A-Sepharose chromatography and preparative sodium dodecyl sulfate polyacrylamide gel electrophoresis. A rabbit antiserum was raised against this purified protein. The specificity of the antiserum was demonstrated by neutralization of rat adrenal gland DBH activity, labeling of rat adrenal medulla on histological sections, and, after Western blot, labeling of the 75,000 molecular-weight band in the different fractions associated with DBH activity during purification. The antiserum had a higher affinity for the sDBH denatured by sodium dodecyl sulfate than for the native protein. It had a higher affinity for sDBH than for mDBH. These results strongly suggest the presence of specific hydrophilic epitopes on the sDBH, revealing structural differences between the two hydroxylase forms. Two protein bands were stained on Western blots of crude rat adrenal gland extract. One band had an apparent molecular weight of 75,000, and the other of 82,000. Our results showed that the two proteins contained similar epitopes, an observation suggesting a close structural relationship. The higher-molecular-weight protein could be the 75,000 protein before covalent modifications and cleavage. PMID- 3047320 TI - Immunocytochemical localization of vasoactive intestinal polypeptide (VIP) in the brain of the little brown bat (Myotis lucifugus). AB - Vasoactive intestinal polypeptide (VIP)-like immunoreactivity has been examined in the brain of the little brown bat, Myotis lucifugus, using light microscopic immunocytochemistry and the indirect antibody enzyme method of Sternberger. Animals were sacrificed at three different and discrete levels of physiological activity: euthermic, hypothermic and hibernating. The density and distribution of immunoreactive neurons and fibres was compared in the three animal groups with the aid of a computerized image analysis system. Our results were compared with those of previous studies in laboratory species such as the rat and cat. Our study has demonstrated marked changes in the density of VIP-immunoreactive fibres and plexuses in the anterior hypothalamic area which correspond to the physiological state of the animal. In addition we have demonstrated the presence of VIP immunoreactive perikarya in a number of previously unreported locations. These include the paraventricular and periventricular hypothalamic nuclei, the linear raphe nucleus, nucleus interfascicularis, and in neurons embedded in the fibres of the dorsal tegmental decussation. PMID- 3047319 TI - Rapid increases in glial fibrillary acidic protein mRNA and protein levels in the copper-deficient, brindled mouse. AB - The brindled mouse (MObr/y) carries an X-linked mutation that produces severe copper deficiency. Affected males suffer profound deficits in oxidative metabolism. We have examined astrocyte pathology in MObr/y during development and have found marked changes in the metabolism of glial fibrillary acidic protein (GFAP). Immunocytochemistry with anti-GFAP antisera revealed a marked increase in staining at postnatal day 12 (P12), compared to heterozygous female and unaffected male littermates, particularly in neocortex and thalamus. Septum, hypothalamus, and striatum showed little change. Western blot analysis revealed increased levels of GFAP in MObr/y forebrain and cerebellum. Levels of GFAP mRNA were determined by Northern blotting with a mouse GFAP cDNA probe. At P10, mRNA levels were normal, but increased to 8-10 times normal by P12. Levels at P15 remained similarly elevated. Thus, immunostaining and protein determinations correlate with mRNA elevations. Astrocytes can alter GFAP mRNA and protein levels over a relatively short time. Counts of neocortical cells did not reveal differences in cell numbers between MObr/y and controls, indicating that the observed changes reflect increased cellular levels and not a large increase in the numbers of astrocytes. PMID- 3047321 TI - Evidence that migratory oligodendrocyte-type-2 astrocyte (O-2A) progenitor cells are kept out of the rat retina by a barrier at the eye-end of the optic nerve. AB - There is evidence that oligodendrocyte-type-2 astrocyte (O-2A) progenitor cells migrate along the developing rat optic nerve from the chiasm toward the eye before differentiating into oligodendrocytes that myelinate the retinal ganglion cell axons in the nerve. Why, then, do these progenitor cells not migrate into the eye, differentiate into oligodendrocytes and myelinate the nerve fibre layer of the retina? Myelination would opacify the neural retina and thereby severely impair vision. Here we provide evidence that there is a barrier at the eye-end of the rat optic nerve that prevents the migration of O-2A progenitor cells into the retina. Our findings in the rat support a previous hypothesis that such a barrier keeps myelin-forming glial cells out of the human retina. PMID- 3047322 TI - Neurite outgrowth from mammalian CNS neurons on astrocytes in vitro may not be mediated primarily by laminin. AB - A number of in vitro studies suggest that certain components of the extracellular matrix (ECM), such as the glycoprotein laminin, can promote neurite outgrowth. In the present study the presence of laminin and heparan sulphate proteoglycan, another ECM molecule, on CNS glial (astrocytes) and non-glial (leptomeningeal) cells in vitro was examined. In addition, the ability of these cells and laminin coated tissue culture substrates to promote neurite outgrowth from developing mammalian cerebellar cortical neurons was also assayed. Leptomeningeal cells were found to be labelled much more intensely with antibodies against laminin and heparin sulphate proteoglycan than were astrocytes. However, the proportion of neurons extending neurites was fourfold greater on astrocyte monolayers than on leptomeninges, and twofold greater than that on laminin. In addition, the length of the neurites growing on astrocyte monolayers was three- to fourfold greater than that on laminin or leptomeninges. Neurite outgrowth on leptomeninges and laminin could not be enhanced by culturing in astrocyte conditioned medium. The ability of various antibodies to block neurite outgrowth on these monolayers was also studied. These results suggest that the robust neurite outgrowth from mammalian CNS neurons plated on astrocytes may be mediated via an astrocyte surface molecule distinct from laminin. PMID- 3047323 TI - Structural components in the synaptic cleft captured by freeze-substitution and deep etching of directly frozen cerebellar cortex. AB - Structural components in the synaptic cleft were examined in cerebellar excitatory synapses by conventional electron microscopy and by rapid freezing followed by freeze-substitution or deep etching. Two transverse components and one parallel element were identified in the clefts of rapidly frozen and freeze substituted synapses: (i) bridging fibrils, 4-6 nm in diameter, that span the cleft; (ii) columnar pegs, 4-6 nm wide and 8-15 nm high, projecting from the postsynaptic surface; and (iii) intervening fine fibrils running parallel to the apposed synaptic membranes. These were more clearly visible in deep-etched synapses, although the postsynaptic pegs were difficult to distinguish from intramembrane particles in the cross-fractured postsynaptic membranes. Deep etching also revealed other fibrils on the cytoplasmic surface of the postsynaptic membrane. These appear to contact the membrane surface or the intramembrane particles. Freeze-substituted materials also displayed the fibrillar components in the postsynaptic dense fuzz, but failed to display the presynaptic dense projections typically observed in thin sections or deep-etched replicas of the conventionally fixed materials. The bridging fibrils are likely to play a mechanical role in holding the synapse together, while the short pegs may be integral parts of the receptor molecules. PMID- 3047324 TI - Development of macroglial cells in rat cerebellum. I. Use of antibodies to follow early in vivo development and migration of oligodendrocytes. AB - The origin of oligodendrocytes and astrocytes in the CNS is still a focus of much experimentation and controversy. We have used antibodies against ganglioside GD3 and galactocerebroside (GC) to follow the origin and development of rat cerebellar oligodendrocytes both in vitro and in vivo. The immunofluorescent identification of GC+ cells in the rat neonatal cerebellum in vivo, revealed that cells initially GD3+/GC- appeared to make the transition via GD3+/GC+ cells to GD3-/GC+ oligodendrocytes. This sequence of events closely paralleled the maturation of cerebellar oligodendrocyte precursors found in serum-free dissociated culture. In contrast, whereas both GD3+ and glial fibrillary acidic protein-positive cells were seen in serum-containing dissociated culture and also in freshly dissociated suspensions of cerebellum at postnatal days 0 to 6, such cells could not be identified in situ. Putative GD3+/GC- oligodendrocyte precursor cells arose from the deeper regions of the cerebellum at birth, perhaps initially from the superior medullary velum adjacent to the fourth ventricle, and appeared to migrate into the developing folia just prior to myelination and the acquisition of GC. PMID- 3047325 TI - Disruption of active zones in frog neuromuscular junctions following treatment with proteolytic enzymes. AB - Frog neuromuscular junction treated with proteolytic enzymes to remove the basal lamina were studied with freeze-fracture techniques in order to examine the influence of the basal lamina in the maintenance of active zone ultrastructure. The active zone is believed to be the site of transmitter release and has a unique membrane organization and location in the neuromuscular junction. After removal of the basal lamina by successive treatment of 0.01% collagenase and 0.1% protease for 1 h each, active zone disruption was observed. Some active zones became segmented, and some were also randomly located and oriented, but they still had normal double-row particle organization. Others contained only clusters of large intramembrane particles. These disorganized active zones were still functional as indicated by the presence of vesicle openings. Some enzyme-treated junctions were also exposed to the membrane cholesterol probe, filipin, to examine the expression of membrane lipid heterogeneity in disrupted active zones. As in normal active zones, filipin-sterol complexes were absent. The densities of background particles in the presynaptic membranes and of large particles thought to be acetylcholine receptors were not significantly altered by the enzyme treatment. Although a direct effect of the enzymes on active zone ultrastructure can not be totally excluded, the present work is consistent with a maintenance role of the basal lamina in active zone organization and location. PMID- 3047326 TI - Immunohistochemical localization of parvalbumin in rat and monkey autonomic ganglia. AB - The cellular distribution of parvalbumin-like immunoreactivity in autonomic ganglia such as superior cervical sympathetic ganglia, paravertebral sympathetic chain ganglia (T6), ciliary ganglia and enteric ganglia was investigated by immunohistochemical peroxidase-antiperoxidase methods using an antiserum against rat skeletal muscle parvalbumin. We detected parvalbumin-like immunoreactivity in almost all neurons of rat superior cervical sympathetic ganglia and other paravertebral sympathetic chain ganglia, where the antigen was located in the cytoplasm but the nuclei were not labelled. No neurons positive for parvalbumin like immunoreactivity were observed in rat ciliary ganglia or enteric ganglia. In monkey, almost all neurons of the superior cervical sympathetic ganglia contained parvalbumin-like immunoreactivity, but none of the neurons of the ciliary ganglia were labelled with the antiserum to parvalbumin. These results suggest that parvalbumin-like immunoreactivity exists in a specific subpopulation of the neurons of the autonomic nervous system. PMID- 3047327 TI - Acquisition of vimentin in astrocytes cultured from postnatal rat brain. AB - Vimentin and glial fibrillary acidic protein (GFAP) represent the principal constituents of intermediate filaments found in astrocytes. In contrast to vimentin-GFAP transition which occurs during glial development in situ, vimentin coexists with GFAP in cortical astrocytes allowed to differentiate in culture. To examine whether culture conditions or proliferative activity of the cells is responsible for the expression of vimentin, we generated cultures of GFAP positive, vimentin-negative astrocytes isolated from 26-day postnatal rat brain cortices. Isolated astrocytes are characterized by a very thin rim of perinuclear cytoplasm and by numerous processes. Antiserum to GFAP labelled major processes and cell somata of some astrocytes, especially those with relatively short and large processes. Within 3 days in culture, all astrocytes accumulated GFAP in hypertrophic cell bodies and many began to express vimentin. Vimentin appeared primarily close to nuclei, and filaments of vimentin extended into proximal segments of the cell processes. In some astrocytes, however, vimentin was always absent. Combined double immunolabelling and histoautoradiography experiments demonstrated that the acquisition of vimentin was independent of the ability of astrocytes to incorporate tritiated thymidine. The results indicate that astrocytes isolated from 26-day postnatal rat brain are heterogeneous with respect to their ability to express vimentin and that vimentin synthesis is not correlated with the growth state of the cells as had been previously suspected. PMID- 3047328 TI - Postsynaptic arch in the frog neuromuscular junction: paramembranous protuberances coating the inner surface of the postjunctional membrane. AB - The frog neuromuscular junction was fixed and processed for electron microscopy according to the method of rapid freezing followed by freeze-substitution. The synaptic structures, including cleft material, paramembranous cytoplasmic coating on the postsynaptic membrane, and subsynaptic cytoplasmic elements, were examined in thin sections. The basal lamina, about 50 nm thick, was seen to bisect a synaptic cleft approximately 100 nm wide. The lamina consists of two parts: the central dense line and the fine filaments protruding from it in the direction of the apposing postjunctional membrane. Present on the cytoplasmic surface of the postjunctional membrane are electron-dense protuberances, 41 +/- 5 nm in width and 27 +/- 5 nm in height (top to membrane centre). They are arranged in regular parallel rows, at 54 +/- 4 nm intervals (centre to centre). The paramembranous protuberance coats the inner surface of the postjunctional membrane at its apex as well as at the middle portion of the junctional process, pointing to its probable hairpin-like course in a transverse plane of the process. From its location and three-dimensional arrangement, this protuberance was termed the 'postsynaptic arch'. A filamentous meshwork is present just beneath the postjunctional membrane and extends into the cell interior. The submembranous meshwork appears to connect to the underlying bundles of cytoskeletal filaments. The possibility is discussed that the postsynaptic, electron-dense arch corresponds to the 43-kDa protein, a major alkaline-extractable protein thought to be associated with the cholinergic receptor molecules in the postsynaptic membrane. PMID- 3047329 TI - Neither laminin nor prior optic nerve section are essential for the regeneration of adult mammalian retinal ganglion cell axons in vitro. AB - Retinal explants obtained from normal adult rats and from operated animals in which the optic nerve had been sectioned 10 days previously were cultured in either serum-containing or serum-free medium on poly-L-lysine and laminin substrata. Regenerating ganglion cell axons growing from these explants have been identified using monoclonal antibodies against Thy-1.1 cell surface glycoprotein and the 200-kDa subunit neurofilament protein. Irrespective of substratum or medium composition, axons regenerated from 28-49% of normal rat retinal explants. This percentage increased to 60-84% of explants from operated rats. There were no significant differences in percentages of explants from normal or operated rats showing neurite outgrowth when substrata of either poly-L-lysine or laminin were compared in serum-free medium. In serum-containing medium the results were less easily interpreted due to the presence of an outgrowth of non-neuronal (glia and mesenchymal) 'flat cells', which served as a preferred axonal substratum in many cases. Thus we show that adult rat retinal ganglion cell axons will regrow in vitro, and that a 'priming' optic nerve section will increase this response. In neither case is the response laminin-dependent. PMID- 3047330 TI - The dose dilemma. PMID- 3047331 TI - More is better. PMID- 3047332 TI - High-dose combination alkylating agents with bone marrow support as initial treatment for metastatic breast cancer. AB - To evaluate the effect of high-dose chemotherapy in the treatment of metastatic breast cancer, we performed a phase II trial of a single treatment with high-dose cyclophosphamide (5,625 mg/m2), cisplatin (165 mg/m2), and carmustine (600 mg/m2), or melphalan (40 mg/m2) and bone marrow support as the initial chemotherapy for metastatic breast cancer. Twenty-two premenopausal patients with estrogen receptor negative, measurable metastatic disease were treated. Twelve of 22 patients (54%) obtained a complete response at a median 18 days. The overall response rate is 73% (complete and partial response). Median duration of response in the patients achieving complete response was 9.0 months with a median duration of survival for complete responders that is currently undefined. Relapse occurred predominantly at sites of pretreatment bulk disease or within areas of previous radiation therapy. Toxicity was frequent and five patients died of therapy related complications. The results indicate that a single treatment with intensive combination alkylating agents with bone marrow support can produce more rapid and frequent complete responses than conventional chemotherapy when used as initial chemotherapy for metastatic breast cancer, although median disease-free and overall survival is not improved. Three patients (14%) remain in unmaintained remission beyond 16 months. PMID- 3047333 TI - Randomized trial of observation versus adjuvant therapy with cyclophosphamide, fluorouracil, prednisone with or without tamoxifen following mastectomy in postmenopausal women with node-positive breast cancer. AB - Following mastectomy for node-positive breast cancer, 261 postmenopausal women were randomized to observation or adjuvant treatment with cyclophosphamide, fluorouracil, prednisone (CFP) alone or combined with tamoxifen (T). Doses used were: C, 150 mg/m2 intravenously (IV) days 1 to 5; F, 300 mg/m2 IV days 1 to 5; P, 10 mg by mouth 3 times daily on days 1 to 7; and T, 10 mg by mouth 2 times daily. A total of ten courses of treatment, administered every 6 weeks, was planned and T was stopped 6 weeks after the last course of CFP. Two hundred thirty-four patients were fully eligible and evaluable. With a median observation time slightly in excess of 5 years, the proportion of recurrences on each arm were: CFP, 29 of 75 (39%); CFPT, 29 of 71 (41%); and observation, 50 of 88 (57%). Relapse-free survival distributions for both CFP and CFPT were superior to observation (both two-sided P = .01). Considering prognostic factors in covariate analysis revealed two-sided P = .0006 for CFP v observation and P = .0003 for CFPT v observation. No substantial difference was identified between CFP and CFPT. Survival data are not yet mature with 31% dead; and, although slight separations of the curves exist in favor of the treatment arms, no significant differences in survival have been seen. Both adjuvant therapy programs are well tolerated and there were no treatment-related deaths. Further maturation of the data is required to determine if the advantages in relapse-free survival will be translated into any overall survival benefit which must be considered the goal of primary interest. PMID- 3047334 TI - A randomized comparison of haloperidol plus dexamethasone versus prochlorperazine plus dexamethasone in preventing nausea and vomiting in patients receiving chemotherapy for breast cancer. AB - The antiemetic effectiveness of haloperidol plus dexamethasone was compared with that of prochlorperazine plus dexamethasone in a prospective study of patients receiving chemotherapy for breast cancer. Chemotherapy consisted of cyclophosphamide, doxorubicin or methotrexate, and fluorouracil in all patients. Patients who received the doxorubicin-containing combination experience significantly more nausea and vomiting than those who received the chemotherapy combination containing methotrexate. There was no significant difference between the two antiemetic regimens in the overall incidence of post-chemotherapy nausea and vomiting, but patients who received haloperidol and dexamethasone did have a lower incidence of severe vomiting. Neither regimen is highly effective, especially for the combination containing doxorubicin, and cannot be recommended as standard antiemetic therapy for this population of patients. PMID- 3047335 TI - Adjuvant chemotherapy with vincristine, cyclophosphamide, and doxorubicin after radiotherapy in local-regional nasopharyngeal cancer: results of a 4-year multicenter randomized study. AB - To evaluate the effect of adjuvant chemotherapy in patients with local-regional nasopharyngeal carcinoma (NPC) (squamous or undifferentiated) in complete remission at the end of curative radiotherapy (RT) 229 patients were randomized from 1979 to 1983 in a multicenter study to no further therapy (116 patients) or a combination of vincristine, cyclophosphamide, and Adriamycin (doxorubicin; Adria Laboratories, Columbus, OH) (VCA) for six monthly cycles (113 patients). The RT and RT + VCA groups were well balanced for median age (50 v 49 years), histology (undifferentiated carcinoma, 73% v 70%), tumor extent (tumor limited to nasopharynx, 57% v 57%), and nodal extent (negative nodes 26% v 24%, nodes in the lower cervical levels, 17% v 16%). RT was delivered to the nasopharynx, the base of the skull, and bilateral cervical nodes using a split course technique over 10 weeks up to the dose of 60 to 70 Gy in involved sites and 50 Gy to negative nodes. Response to RT was evaluated within 65 days post-RT treatment. Analysis at 48 months did not show significant difference between the two treatment groups in terms of relapse-free survival (RT, 55.8%, RT + VCA, 57.7%, P = .45) and overall survival (RT, 67.3%, RT + VCA, 58.5%, P = .13). The pattern of relapse was similar in the two treatment arms. Distant metastases were the cause of treatment failure in about 50% of relapsing patients. Although the results of the present study did not show any benefit from VCA administered after curative RT, combined systemic chemotherapy should be further explored due to the high incidence of local and distant failure after intensive RT. PMID- 3047336 TI - Androgen priming and chemotherapy in advanced prostate cancer: evaluation of determinants of clinical outcome. AB - We conducted a randomized clinical trial in men with stage D2 prostate cancer to test whether androgen priming potentiates the efficacy of cytotoxic chemotherapy. Eighty-five men with progressive prostate cancer refractory to orchiectomy were treated continuously with aminoglutethimide and hydrocortisone to lower adrenal androgen secretion and were administered cyclic intravenous (IV) chemotherapy. The patients were randomized to receive either androgen priming or no additional treatment for three days before and on the day of chemotherapy. Median duration of follow-up was 43 months. Response rate (remission plus disease stabilization) was not significantly different between the stimulation and control arm when the analysis was restricted to evaluable patients (79% v 73%, respectively) or when it was extended to all patients (46% v 61%). Median duration of response was similar for the stimulation and control arm (9 and 10 months, respectively). Median survival was 10 months in the stimulation and 15 months in the control group (P = .0047). The androgen sensitivity of the tumors was supported by the greater toxicity in the stimulation arm associated with androgen administration. Factors found to be independently associated with improved clinical outcome included a high Karnofsky score and hematocrit, long duration of response to the initial castration, and normalization of an elevated serum acid phosphatase on treatment. We conclude that in this group of patients with advanced disease, androgen priming does not potentiate the efficacy of chemotherapy and is actually associated with a worse outcome. Furthermore, our data emphasize the heterogeneity of biologic behavior of prostate cancer. PMID- 3047337 TI - Brain metastases in adenocarcinoma of the lung: frequency, risk groups, and prognosis. AB - A consecutive group of 259 patients with inoperable adenocarcinoma of the lung (ACL) were observed to define risk groups for and frequency of brain metastases together with prognosis. All patients received chemotherapy in a three-armed randomized trial. Brain metastases were diagnosed in 25 patients before protocol entry and in 37 during treatment. Brain autopsy was performed in 87 patients and was positive in 38 (44%). Eleven of these (29%) were not diagnosed clinically. Patients younger than 60 years had a somewhat higher overall frequency of brain metastases than older patients. Patients with initial performance status above 60% and patients responding to chemotherapy had higher risk for developing brain metastasis during treatment than other patients, probably because of the increasing cumulated risk for this complication with prolonged survival. Median survival after onset of brain metastases was 73 days and survival was significantly shorter for these patients than for patients without this complication at days 0, 90, 180, and 365 after protocol entry. Thus, brain metastases is a frequent complication in ACL and the frequency increases with prolonged survival. Survival after development of brain metastases is short and it is questionable whether the inclusion of this subgroup of ACL patients into experimental cytostatic treatments is justified. PMID- 3047338 TI - Vincristine and prednisone prolong the survival of patients receiving intravenous or oral melphalan for multiple myeloma: Cancer and Leukemia Group B experience. AB - A total of 589 patients with previously untreated multiple myeloma were randomized to receive daily oral melphalan, pulse-dose intravenous (IV) melphalan, carmustine (BCNU), or lomustine (CCNU). All patients received an initial tapering course of prednisone (Pred). During week 22 (day 154), patients were randomized to receive or not to receive additional therapy with vincristine (VCR) (1 mg/m2) and prednisone (0.6 mg/kg/d for seven days) at 8-week intervals. The influence of VCR/Pred was determined in 302 patients who remained on study beyond 22 weeks after initial therapy. VCR/Pred converted a significant percentage of nonresponders to responders in patients treated with melphalan (55% v 19%, P = .002), but not in patients treated with a nitrosourea (48% v 23%, P = .06). Survival beyond week 22 was significantly longer following the addition of VCR/Pred in patients receiving melphalan (median, 35.3 months v 27.0 months; P = .003) but not in patients receiving BCNU or CCNU (median, 28.1 months v 26.2 months; P = .91). These differences were seen both for oral and IV melphalan. A trend for beneficial effect of VCR/Pred was definitely seen in the good-risk patients (P = .03) but only suggestive for poor-risk patients (P = .12). Following adjustment for VCR/Pred effects, there were no differences in the survival of patients receiving any of the four initial treatments. PMID- 3047339 TI - Adjuvant chemotherapy for patients with high-grade soft-tissue sarcomas of the extremity. AB - We have previously reported the results of a randomized trial that demonstrated the survival benefit of adjuvant chemotherapy in the treatment of patients with high-grade extremity sarcomas compared with no chemotherapy. This regimen included doxorubicin, cyclophosphamide, and methotrexate. This report updates and extends our experience. The median follow-up of this trial is now 7.1 years and reveals a 5-year disease-free survival of 75% and 54% for chemotherapy and no chemotherapy groups, respectively (two-sided P [P2] = .037). The 5-year overall survival for patients in this trial was 83% and 60% for the chemotherapy and no chemotherapy groups, respectively, with a trend towards improved survival in the chemotherapy arm (P2 = .124). Because of doxorubicin-induced cardiomyopathy we performed a subsequent randomized trial comparing this high-dose regimen to reduced cumulative doses of doxorubicin and cyclophosphamide without methotrexate. Eighty-eight patients were entered into this trial which has a median follow-up of 4.4 years. The 5-year disease-free and overall survival for patients treated with the reduced doses of chemotherapy was 72% and 75%, respectively, and was not significantly different from the high-dose regimen. No patients developed congestive heart failure on this study. We conclude that adjuvant chemotherapy improves disease-free survival in patients with extremity soft-tissue sarcomas. The overall survival advantage in patients receiving adjuvant chemotherapy in our initial randomized high-dose chemotherapy trial has diminished though it continues to favor the chemotherapy group. A reduced-dose chemotherapy regimen was found to be comparable to the high-dose regimen. PMID- 3047340 TI - Dose-response in the treatment of breast cancer: a critical review. AB - In animal tumor models the dose-response curve for cytotoxic agents, especially cyclophosphamide, may be steep, but the slope and shape of this curve depends not only on the drug used but on the schedule of drug administration, the specific tumor type, tumor cell kinetics, and tumor mass. It might be anticipated from these studies that the human tumors most sensitive to dose effects would be leukemia, lymphoma, small-cell carcinoma of the lung, and testicular tumors rather than the low growth fraction, relatively less responsive tumors such as breast cancer. However, the clinical evidence for a steep dose-response curve in any tumor type is limited. For breast cancer such evidence is largely retrospective or derived from uncontrolled trials. The data available from randomized trials makes it seem unlikely that small, or even moderate, reductions in drug dose for nontrivial reasons will compromise the survival of patients with either early or metastatic disease. In spite of promising data from small trials, there is, as yet, inadequate evidence to justify the use of very-high-dose therapy and autologous marrow transplant outside the setting of a well-designed clinical trial. The value of high-dose therapy, intensive dose rate, and cumulative drug dose should each be studied in randomized controlled trials. PMID- 3047342 TI - Congenital anaplastic astrocytoma with favorable prognosis. Case report. AB - A large intracranial tumor that caused macrocrania leading to dystocia was demonstrated by prenatal ultrasound examination. After birth, computerized tomography (CT) confirmed the presence of a giant supratentorial tumor with a large cyst. When the infant was 20 days old, the tumor was radically extirpated. Neuropathological examination revealed an astrocytoma with focal signs of anaplasia showing a macrocyst as well as multiple microcysts resulting from hemorrhages into the tumor. Although no adjuvant radio- or chemotherapy was administered, the child had nearly normal psychomotor development without clinical or CT evidence of tumor recurrence, and is now 3 years old. PMID- 3047341 TI - Intra-arterial bromodeoxyuridine radiosensitization and radiation in treatment of malignant astrocytomas. AB - Bromodeoxyuridine (BUdR), a nonhypoxic radiosensitizing drug, is a halogenated pyrimidine analog that is incorporated into the deoxyribonucleic acid of dividing cells in a competitive process with thymidine; BUdR also sensitizes these cells to radiation therapy. Neurons and glial cells have a very low mitotic rate. They will not incorporate BUdR and will not be sensitized. Bromodeoxyuridine is best delivered intra-arterially because of its regional advantage, calculated to be between 6 and 16. An 8-week BUdR infusion is delivered before and during radiation therapy through a permanently implanted pump with a catheter placed retrograde into the external carotid artery. Eighteen patients with malignant glioma (15 grade IV, and three grade III) were entered into a Phase I dose escalation protocol with BUdR dosages ranging from 400 to 600 mg/sq m/day. The maximum dose that can be tolerated appears to be 400 mg/sq m/day for 8 weeks. The 18 patients entered in this study have a median Kaplan-Meier estimated survival time (+/- standard error of the mean) of 22 +/- 5 months with 11 patients still alive. Three patients are alive at 30, 29, and 21 months after diagnosis with no evidence of tumor on computerized tomography. There have been no vascular complications. Side effects in all patients have included anorexia, fatigue, ipsilateral forehead dermatitis, blepharitis, iritis, and nail ridging. Myelosuppression requiring dose reduction occurred in one patient. One patient had a Stevens-Johnson syndrome requiring termination of BUdR. It is concluded that intra-arterial BUdR may improve survival times in patients with malignant gliomas. PMID- 3047343 TI - Epidural hibernoma as a complication of corticosteroid treatment. Case report. AB - Centripetal fat deposition is a well-recognized consequence of excessive use of corticosteroids, either endogenous or exogenous. Recently, several patients receiving large doses of corticosteroids have suffered compressive myelopathies due to excessive epidural fat collections, labeled "epidural lipomatosis." Two of these have been children, and a third child is reported here. This child was receiving chronic steroids for juvenile rheumatoid arthritis when he presented with such a myelopathy, which was confirmed by metrizamide computerized tomography myelography as well as by surgical exploration. Histological examination revealed that the epidural tissue was a brown-fat tumor or "hibernoma." An epidural hibernoma has not been described previously. The histological and endocrine features of fat in Cushing's syndrome are discussed, and the literature concerning hibernoma and epidural lipomatosis is reviewed. PMID- 3047344 TI - Increased insulin sensitivity in iron-deficient rats. AB - Iron deficient (ID) and control (C) rats were studied to determine if severe iron deficiency alters insulin-stimulated glucose disposal. Euglycemic hyperinsulinemic glucose clamps were conducted by infusing insulin (2 m mu.kg 1.min-1, constant rate) for 120 min while maintaining euglycemia. In a 12-h fasted state, ID rats were hyperglycemic (109.4 +/- 4.0 mg.dL-1 arterial plasma glucose, x +/- SEM) when compared with C rats (86.9 +/- 3.4 mg.dL-1) (P less than 0.05). Even though insulin was infused identically on a per kilogram body weight basis for both groups, the resulting hyperinsulinemia was higher in ID rats (3.1 +/- 0.27 ng.mL-1) compared with C rats (2.3 +/- 0.4 ng.mL-1) at the end of the clamp. Glucose infusion rates required to maintain euglycemia were twofold higher in ID rats (27.0 +/- 5.4 mg.kg-1.min-1) versus C rats (13.1 +/- 3.3 mg.kg-1.min 1) (P less than 0.05). Circulating lactic acid increased in both groups, and the concentrations in ID rats (3.2 +/- 0.4 mmol.L-1) were significantly higher than those in C rats (1.8 +/- 0.5 mmol.L-1) at the end of the clamp. When the efficiency of insulin to dispose glucose was evaluated by calculating the glucose disposal divided by the prevailing insulinemia, ID rats could dispose of almost twice the glucose per unit of insulin [9.0 +/- 0.6 (mg.kg-1.min-1)/(ng.mL-1)] when compared with C rats [5.6 +/- 0.9 (mg.kg-1.min-1)/(ng.mL-1)] (P less than 0.05). The data indicate that insulin sensitivity is increased in ID rats and that ID rats cannot metabolize exogenous insulin as well as C rats. PMID- 3047345 TI - Transcriptional regulation of carnitine palmitoyltransferase synthesis in riboflavin deficiency in rats. AB - Riboflavin deficiency leads to depressed mitochondrial fatty acid oxidation rates but increased activity of carnitine palmitoyltransferase (CPT). Starvation leads to increased CPT activity in ad libitum-fed, riboflavin-supplemented rats. The present studies examined the mechanism of the increase in CPT activity in riboflavin deficiency and whether it was additive to that seen in starvation. Rats were divided into three groups initially: riboflavin-sufficient, ad libitum fed; riboflavin-deficient, ad libitum-fed; and pair-fed. These groups were subdivided after 5 wk into fed and 24- and 48-h starved groups. When riboflavin deficient rats were starved for 24 or 48 h, there was only a 30-40% increase in hepatic CPT activity, in contrast to the ad libitum-fed, riboflavin-supplemented rats, in which activity increased twofold. CPT activity of pair-fed rats was similar to that of controls in the fed state and did not increase significantly with starvation. CPT translation, mRNA levels and transcription rates correlated with CPT activity, as did immunoreactive CPT. Concurrently, hepatic ketone production and plasma beta-hydroxybutyrate concentration increased during starvation in the control and pair-fed but not in the riboflavin-deficient rats. The results indicate that increased CPT activity in riboflavin deficiency and starvation results at least in part from increased synthesis. Furthermore, the data support previous work suggesting that the block in fatty acid oxidation occurs in the beta-oxidation pathway at the level of acyl-CoA dehydrogenases. PMID- 3047346 TI - AIDS and chemical dependency: an overview. PMID- 3047347 TI - AIDS: perceptions versus realities. PMID- 3047348 TI - AIDS and drug use: an international perspective. PMID- 3047349 TI - The intravenous drug user and secondary spread of AIDs. PMID- 3047350 TI - Psychological assessment and AIDS research with intravenous drug users: challenges in measurement. AB - The instruments used for psychological assessment have been under close scrutiny for many years. In particular, ethnic and racial minorities have pointed out that misapplication of instruments standardized to White middle-class norms can result in incorrect assessments. An analogous situation exists with IVDUs. In the work of the present authors with IVDUs, they were found to be a very diverse group. Contrary to common wisdom, they differ by race, ethnicity, age, and drug use profiles. However, their economic circumstances and social stigma make them a special case in terms of psychological assessment. Given the unique characteristics of IVDUs, it behooves researchers to carefully examine the standardized instruments that are available for psychological evaluation. Too often, measures standardized on White middle-class samples lack the value neutrality that makes them applicable across disparate groups. In addition, many such measures are designed with certain presumptions that do not necessarily hold true with this population (e.g., willingness and/or ability to communicate intimate information about one's feelings and psychological states). This article briefly describes some of the challenges encountered in examining standardized instruments for use in the study of IVDUs, their health psychology and AIDS related behavior. Concerns with self-report biases, literacy, attentional focus, measurement constructs, and drug states confounding psychological states all pose challenges to psychological research with this heterogeneous population. While the need for direct intervention on the sexual and needle-sharing behaviors of IVDUs remains paramount in the combat against the spread of AIDS, researchers must also continue with the further development of basic measurement tools.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3047351 TI - GM-CSF produced by recombinant vaccinia virus or in GM-CSF transgenic mice has no effect in vivo on murine cutaneous leishmaniasis. AB - The hemopoietic growth and differentiation regulators, granulocyte-macrophage colony-stimulating factor (GM-CSF) and the multipotential stimulating factor (multi-CSF) have been shown to have major effects on the effector function of mature macrophages. In this study we have examined the effect of recombinant GM CSF and multi-CSF expressed transiently from recombinant vaccinia virus, or constitutively in GM-CSF transgenic mice on the development of cutaneous leishmaniasis, caused by Leishmania major in genetically susceptible or resistant mice. We observed no effect on the development of lesions when GM-CSF or multi CSF were administered before infection, nor on the healing of lesions when they were administered after appearance of lesions. Although only some of the GM-CSF transgenic mice or their normal littermates developed lesions after infection with L. major, there was no difference between the groups in the rate of lesion development or in the size of lesions. PMID- 3047352 TI - Growth of pleomorphic Trypanosoma brucei rhodesiense in irradiated inbred mice. AB - It was shown that irradiation (650 rad) of 7 inbred strains of mice did not block the ability of Trypanosoma brucei rhodesiense to transform from the long slender (LS) to the short stumpy (SS) form or alter the plateau in parasitemia. In addition, it was observed that significant differences in parasitemia levels, in the rate of transformation from the LS to the SS form, as well as in the survival times occurred between the irradiated C3HeB/FeJ and several of the other strains. These differences in the nonspecific ability to control parasitemia appeared to be characteristic for each inbred strain of mice. The resistant strains generally had lower parasitemia than the susceptible strains. However, it was also shown that there is not a one-to-one correlation between the innate ability of a mouse strain to control its initial parasitemia, and the strain's ability to clear the parasitemia or increase its survival time. It was therefore concluded that the hypothesis which states that the ability of an animal to increase nonspecifically the rate of transformation, and therefore to lower the parasitemia, allowing intact animals to respond immunologically and survive longer is either incorrect or incomplete. The results further show that the ability of mice to clear their initial parasitemia by an antibody response is not necessarily correlated with their survival time. Therefore, this study suggests that factors other than an antibody response and the nonspecific control of parasitemia are important in resistance. PMID- 3047353 TI - Fatal Caryospora bigenetica (Apicomplexa: Eimeriidae) infections in cotton rats, Sigmodon hispidus. AB - Four groups of cotton rats, Sigmodon hispidus, were shown to be suitable secondary hosts for the viperid coccidium, Caryospora bigenetica, following oral inoculation of a mixture of oocysts and sporocysts. Swelling of the face, ears, and scrota and hemorrhagic ears were the predominant clinical signs and some cotton rats died in 3 of 4 experiments. Developmental stages of C. bigenetica were found in connective tissue components of the ear, nose, cheeks, anal skin, scrotum, and penile sheath of all cotton rats in which these tissues were examined. Additionally, developmental stages of C. bigenetica were found in connective tissue components of the following tissues examined from some cotton rats: tongue, lung, testicle, epididymis, rectum, base of the tail, footpad, and bone marrow. The present study shows that C. bigenetica can be pathogenic for cotton rats and demonstrates many new anatomic sites for developmental stages of this parasite in the secondary host. PMID- 3047355 TI - Nutritional factors determine germ tube formation in Candida albicans. AB - Following a short (3 h) period of carbon starvation, exponential phase yeast cells of Candida albicans rapidly (T50 45 min) formed germ tubes in a glucose/ammonium ion solution. The presence of both a sugar (glucose, sucrose or galactose) and a nitrogen source (ammonium ion or an amino acid metabolized via glutamate) was critical for morphogenesis. PMID- 3047354 TI - The spiny rat louse, Polyplax spinulosa, as a parasite of the rice rat, Oryzomys palustris, in North America. AB - The spiny rat louse, Polyplax spinulosa, was collected, as adults and embryonated ova from the rice rat, Oryzomys palustris, a cricetid rodent, in Davidson Co., Tennessee. This sucking louse is typically parasitic on domestic rats, which are murid rodents. Because most sucking lice are normally host specific, such cross familial host infestation is noteworthy. PMID- 3047357 TI - Late results following esophagomyotomy in children with achalasia. AB - Twenty one children with achalasia of the esophagus were treated from 1970 to 1986. There were 11 girls and ten boys (average age, 10.9 years; range, 6 months to 16 years). Diagnosis was established by barium swallow in 21 cases and confirmed by manometrics and motility studies in 14. Four children had unsuccessful dilatation (range, 1 to 16 dilatations/pt). All 21 children underwent modified anterior Heller esophagomyotomy (transabdominal in 15 and transthoracic in six). Concomitant Nissen fundoplication was performed in three. Follow-up from 1 to 14 years (mean, 6.3 years) showed complete relief of obstruction in 18 patients (86%), while three required additional procedures for persistent dysphagia. One child improved after a single dilatation, but two others eventually required a second esophagomyotomy. Three additional patients subsequently developed gastroesophageal reflux (GER), and two were managed with Nissen fundoplication; the third responded to medical management. The mortality for this series was zero. Postoperative complications occurred in nine children (42%) and was due to atelectasis and postoperative fever. Modified Heller esophagomyotomy is safe and effective in children with achalasia (mortality, 0%; relief of obstruction, 86%). Results were similar after a transabdominal or transthoracic approach. Esophageal dilatation was not an effective method of treatment. Although postsurgical barium swallow showed relief of obstruction, abnormal esophageal motility persisted, suggesting that long-term follow-up is important. PMID- 3047356 TI - Influence of growth conditions on Candida albicans adhesion, hydrophobicity and cell wall ultrastructure. AB - The effect of cultivation in 13 media (10 complex, and three synthetic), as well as altering growth conditions, on Candida albicans adhesion, cell surface hydrophobicity and cell wall ultrastructure was studied. Adhesion of C. albicans to buccal epithelial cells (BECs) was significantly modified by all of the factors tested, particularly growth medium. In general, optimal adhesive activity for C. albicans was observed when the cells were grown in defined media (depending on the carbohydrate used) and/or at 25 degrees C. Moreover, significant differences in adhesion to BECs were noted when C. albicans was grown in the same complex medium from different manufacturers and in different batches of medium from the same manufacturer. Electron microscopy revealed significant differences in surface topography and cell wall ultrastructure of C. albicans grown in different media but none of these differences, including presence or absence of an outer floccular layer, appeared to correlate with the adhesive changes noted, which raises questions regarding the location and nature of the Candida adhesin(s). Likewise, cell surface hydrophobicity could not be correlated with adhesion to BECs but may have influenced yeast coadhesion. The results indicate that Candida adhesion is highly dependent upon the cultivation conditions of the yeast cells tested, and may explain discrepancies in the literature regarding the biochemical nature of the surface component(s) responsible for C. albicans adhesion. PMID- 3047359 TI - Post-necrotizing enterocolitis strictures presenting with sepsis or perforation: risk of clinical observation. AB - Intestinal stenosis or stricture occurs in approximately one third of medically treated infants surviving the acute phase of necrotizing enterocolitis (NEC). Identification of these lesions by the use of routine contrast enemas has been advocated as a means of decreasing potential morbidity from delayed diagnosis. However, the significant incidence of spontaneous resolution and reluctance to submit asymptomatic infants to contrast enema have led recent researchers to reserve these studies for patients developing symptoms of obstruction during a period of close observation. From July 1984 to July 1986, symptomatic strictures developed in five infants (15%) responding to medical management at our institution. Contrast enemas were not routinely performed and four (80%) of these patients presented with life-threatening sepsis or perforation associated with intestinal obstruction. Two infants developed complete colonic obstruction 4 and 6 weeks after discharge from the Intensive Care Nursery, having initially tolerated oral feedings. Both infants were critically ill due to perforation or sepsis and underwent emergency colostomy at community hospitals. Two other infants developed abdominal distension with sepsis and cardiopulmonary decompensation while remaining hospitalized for prematurity and pulmonary insufficiency. These patients became symptomatic 5 and 7 weeks after cautious refeeding while closely monitored in the Intensive Care Nursery. The occurrence of such life-threatening complications suggests that clinical observation alone is not adequate in the management of many of these infants. Contrast enemas should be performed to identify those patients at risk of such potential morbidity or mortality, especially those infants not residing near pediatric surgical facilities. PMID- 3047358 TI - The usefulness of open-lung biopsy in the pediatric bone marrow transplant population. AB - From October 1976 to October 1986, 126 children had bone marrow transplants at the Children's Hospital of Philadelphia. The indications were acute lymphocytic leukemia (ALL) (30), nonlymphocytic leukemia (24), aplastic anemia (15), solid tumors (47), and miscellaneous conditions (10). Of these, 21 (17%) underwent 22 open-lung biopsies. Fourteen of these patients showed no causative microorganism. When a cause was found it was viral (usually cytomegalovirus [CMV]) in three, fungal in one, Pneumocystis carinii alone in two, both viral and pneumocystis in one, and a combination of viral, bacterial, and pneumocystis in one. Thirteen patients died due to continued deterioration after the biopsy. In only two patients was there a significant change in antimicrobial therapy as a result of the biopsy. Both had Pneumocystis (one in combination with virus and bacteria). One patient with chronic infiltrates showed a lymphocytic interstitial pneumonia, which responded well to steroids. Open-lung biopsy is currently of limited value in this patient population. Survival is dismal unless the patient has Pneumocystis. We believe that prospective studies should be set up to compare open-lung biopsy with empiric antimicrobial therapy. A major emphasis must be on prevention. PMID- 3047361 TI - Parents' grief reaction to the diagnosis of their infant's severe neurologic impairment and static encephalopathy. PMID- 3047360 TI - Cervical teratomas: an analysis. Literature review and proposed classification. AB - Cervical teratomas are uncommon lesions usually diagnosed at birth but occasionally reported in older children and adults. During a 58-year span, nine cervical teratomas were identified at our institution (four previously reported): three stillborns with giant tumors; five live newborns; and one adult with a malignant tumor. Of the five newborns, two prematures died within one hour of birth. Of the three survivors, 2 had respiratory distress at birth. These infants were treated with early excision and are well at 7, 6, and 2 years of age. The last patient also had cystic fibrosis. The adult died of metastatic disease 8 months after resection. A literature review disclosed 212 cases in addition to the five reported here. Previous attempts at categorizing cervical teratomas have failed to address clinical patterns and have little prognostic value. We propose a classification based on birth status, age at diagnosis, and the presence or absence of respiratory distress. Group I--stillborn and moribund live newborns: number (N), 27; mortality (M), 100%. Group II--newborn with respiratory distress: N, 99; M, 43.4%. Group III--newborn without respiratory distress: N, 37; M, 2.7%. Group IV--children age 1 month to 18 years: N, 31; M, 3.2%. Group V--adults: N, 23; M, 43.5%. Twenty-six patients in group II and one in group III died without excision of the mass. Seventy-three patients in group II, 36 in group III, and 31 in group IV had extirpation of the tumor. Operative mortality was 11%, 0%, and 3.2%, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3047363 TI - Social support and social networks: a review of the literature. PMID- 3047362 TI - Bone marrow transplant specialty nursing orientation. PMID- 3047364 TI - Cocaine inhibits muscarinic cholinergic receptors in heart and brain. AB - (-)-Cocaine inhibits M2 muscarinic cholinergic binding measured with [3H]quinuclidinyl benzilate in heart and brain with a Ki of 18.8 microM. The cyclic nucleotide 5'-guanylylimidodiphosphate does not shift the competition curve, suggesting that (-)-cocaine is an antagonist. (-)-Cocaine also reverses the methacholine-induced inhibition of guinea pig atrial contractions at a similar concentration. Although (+)-cocaine is about 8-fold more potent than (-) cocaine, (+)-cocaine is not present in extracts of the coca plant. Of the many compounds tested, only (-)-cocaine and lidocaine have a higher affinity at M2 muscarinic receptors than at M1 receptors; other compounds such as (+)-cocaine, norcocaine, procaine and dimethocaine are equipotent at the M1 and M2 subtypes. These results indicate that cocaine can act as an antimuscarinic agent, particularly at higher, toxic doses. PMID- 3047365 TI - Ocular ketone reductase distribution and its role in the metabolism of ocularly applied levobunolol in the pigmented rabbit. AB - The distribution of ketone reductase activity in the anterior segment tissues of the pigmented rabbit eye and its influence on the ocular metabolism of topically applied levobunolol were studied. A reversed phase high-performance liquid chromatography procedure was used to assay for this drug and its metabolite, dihydrolevobunolol. Ocular ketone reductase activity was 3 to 4 times more dependent on NADPH than on NADH. The rank order of activity was corneal epithelium greater than iris-ciliary body greater than conjunctiva greater than lens. No activity was detected in the tears, corneal stroma, sclera or aqueous humor. Ketone reductase activity was entirely cytosolic. The pigmented rabbit was significantly less active than the albino rabbit in ketone reductase. Its activity in the corneal epithelium, iris-ciliary body and lens was most sensitive to inhibition by quercetin, whereas that in the conjunctiva was most sensitive to metyrapone. The ketone reductase in the corneal epithelium contributed more to the metabolism of topically applied levobunolol than its counterpart in the iris ciliary body and lens. Moreover, the extent of levobunolol metabolism both during and after corneal penetration was dose-dependent. Overall, these findings indicate that ocularly applied drugs containing the ketone functional group are subject to varying degrees of metabolism by NADPH-dependent ketone reductases in the corneal epithelium, iris-ciliary body and lens. PMID- 3047366 TI - Cardiovascular actions of the primate-selective renin inhibitor, A-62198. AB - A-62198 [dimethylacetyl-Phe-His-NHCH(cyclohexylmethyl)CH-(OH)C H(OH)CH2N3] is a potent, selective inhibitor of primate renin. This compound induced a dose dependent fall in mean arterial blood pressure (MAP) when administered as an i.v. bolus to anesthetized, salt-depleted monkeys. Both the magnitude and the duration of the hypotensive effect were dose related. Its actions were also studied during acute infusions in anesthetized anephric, normal and salt-depleted monkeys. MAP, heart rate and plasma renin activity (PRA) were determined during baseline and 30 min infusions of vehicle alone, followed by A-62198 as boluses of 0.01, 0.1 and 1.0 mg/kg, each maintained by infusing one-tenth of the bolus dose per minute. Vehicle did not alter base-line values. In the normal monkeys, A-62198 induced a dose-related fall in MAP which achieved statistical significance only at the highest dose, while maximally suppressing PRA at all doses (P less than .05, compared to vehicle). The salt-depleted monkeys responded with a dose-related fall in MAP and inhibition of PRA at all doses (P less than .05, compared to vehicle). A-62198 was relatively ineffective in the anephric monkeys which, as expected, had exceedingly low levels of PRA. Heart rate was unaltered regardless of dose or treatment group. Finally, infusion of 1.0 mg/kg bolus + 0.1 mg/kg/min of A-62198 had no effect on MAP or PRA in 2 kidney-1 clip rats, although MAP was reduced subsequent to a superimposed bolus of 0.1 mg/kg of captopril. We conclude that the renin inhibitor, A-62198, is an effective, primate selective hypotensive agent.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3047367 TI - Evidence for co-transport of sodium, potassium and chloride in mouse pancreatic islets. AB - 1. The presence of a loop diuretic-sensitive co-transport system for Na+, K+ and Cl- was tested in isolated pancreatic islets. 2. Substitution of Cl- with the impermeant anion isethionate or addition of frusemide both reduced the ouabain resistant islets uptake of 86Rb+ (K+ marker) without affecting the ouabain sensitive uptake or equilibrium content of 86Rb+. The effects of Cl- substitution and frusemide were overlapping. 3. D-Glucose reduced the ouabain-resistant islets uptake of 86Rb+. This effect was additive to the effect of Cl- substitution or frusemide. 4. Substitution of Cl- with isethionate or addition of frusemide both reduced the efflux of 86Rb+ from the islets. These effects were additive to the reduction of 86Rb+ efflux induced by D-glucose. 5. Substitution of K+ or Na+ with choline reduced the equilibrium content of 36Cl- in the pancreatic islets. 6. These data are compatible with the operation in the pancreatic beta-cells of a loop diuretic-sensitive co-transport system for Na+, K+ and Cl-, that may serve as an inwardly directed Cl- pump. PMID- 3047369 TI - Early diagnosis of inflammatory complications in human heart recipients using monitoring of peripheral blood cells (CIM). PMID- 3047370 TI - Tumours of the parapharyngeal space. PMID- 3047368 TI - Excitatory amino acids in synaptic excitation of rat striatal neurones in vitro. AB - 1. Intracellular recordings were made from rat striatal neurones in vitro. The cells had resting membrane potentials greater than -60 mV and action potentials greater than 70 mV with spike overshoot of 10-30 mV. 2. In the presence of bicuculline intrastriatal stimulation evoked an excitatory postsynaptic potential (EPSP). The relationship between EPSP amplitude and membrane potential was not linear. The EPSP decreased in amplitude and duration for values of membrane potential more negative than -80 mV and increased in amplitude and duration for values of membrane potential more positive than -50 mV. 3. The mean reversal potential for the EPSP recorded with electrodes filled with potassium methyl sulphate was -9.2 +/- 1.7 mV (mean +/- S.E.M.) in presence of bicuculline (30 microM). A similar reversal potential was obtained with CsCl-filled electrodes. 4. The endogenous broad-spectrum excitatory amino acid antagonist, kynurenic acid (100-500 microM), reduced the EPSP in a dose-dependent way, maximally by 80% at 500 microM, but a residual depolarization remained even at high antagonist concentrations. This effect was associated sometimes with a membrane depolarization and an increase in input resistance. 5. In normal artificial cerebro-spinal fluid solution and at resting membrane potential the specific N methyl-D-aspartate (NMDA) antagonist, (D,L)-2-amino-7-phosphonoheptanoic acid (([D,L)-AP7), did not affect the EPSP amplitude. However, this antagonist partially reduced the EPSP amplitude when the membrane was depolarized beyond -50 mV by intracellular current injection. 6. The nicotinic cholinergic antagonist mecamylamine (10 microM) caused a partial (24 +/- 3%) reduction of EPSP amplitude at resting potential in normal medium. However, in the cells where a reduction of EPSP amplitude was observed it was always accompanied by membrane depolarization (7.1 +/- 2.1 mV). (+)-Tubocurarine and hexamethonium were without effect at 10 microM. 7. When Mg2+ was removed from the bathing solution, the EPSP increased in amplitude (89 +/- 9.5%) and duration. In Mg2+-free medium at resting membrane potential (D,L)-AP7 (30 microM) partially reduced EPSP amplitudes (59 +/- 2.5%). 8. It is proposed that a major component of the EPSP evoked by intrastriatal stimulation is mediated by excitatory amino acids. At resting membrane potential and in normal medium only non-NMDA receptors seem to contribute to the synaptic depolarization, but at depolarized potentials and in Mg2+-free medium an NMDA receptor-mediated component of the EPSP can be demonstrated. PMID- 3047371 TI - Quality control in surgical training--do we need higher examinations? PMID- 3047372 TI - [Cerebral cysticercosis. Diagnostic value of x-ray computed tomography. Apropos of 117 cases]. AB - 117 cases of cerebral cysticercosis in La Reunion are reported. Different CT features are related and situated with the evolutive state of the parasite. CT scan provided accurate diagnostic information except in some intraventricular or subarachnoid location where NMR study seems already to be superior. PMID- 3047373 TI - [Pseudotumor focal xanthogranulomatous pyelonephritis in adults. Role of imaging technics in the diagnosis]. AB - Based on 8 personal cases, pseudo-tumoral xanthogranulomatous pyelonephritis is reviewed with emphasis on diagnosis. The condition is a particular form of chronic renal suppuration of histologic definition (combined lesions of chronic pyelonephritis and xanthogranulomatous foam cells). Two forms are recognized: one diffuse, fairly frequent form corresponding to a pyonephrosis, and a pseudo tumoral focal form, the only type discussed in this report, which raises the problem of diagnosis of a renal mass that requires the application of all currently available exploration means to define its true nature. Intravenous urography, ultrasound and CT scan imaging show a non-specific mass of variable character. Selective renal arteriography sometimes shows inflammatory type vascularization, a valuable aid but again non-specific. Puncture biopsy has been used by few authors. However, the presence of a renal mass associated with a chronic pyelonephritis, lithiasis and recurrent episodes of urinary infection should suggest the diagnosis and make use of imaging techniques to detect the affection and adapt therapy, major oncologic surgery being of no utility. Perhaps NMR imaging will provide a step forward in tissue characterization, but it is too early to say. PMID- 3047374 TI - [The new LEM caval filter in the prevention of pulmonary embolism. Preliminary results of a French multicenter study]. AB - Preliminary results are reported of a prospective multicenter trial of a new caval filter (LEM*) implanted by the percutaneous jugular route in 100 patients, 55 men and 45 women, mean age 67 +/- 13 years, to produce partial interruption of inferior vena cava (IVC). Of the 100 attempts to insert the LEM* filter, 2 failures to catheterize the jugular vessel were reported, 98 filters being placed in the IVC with 82 implantation considered adequate. Of the remaining 16 cases, the filter was inclined (7 cases) or incompletely open (9 cases) with total lack of success in 3 cases. Overall efficacy was obtained therefore in 95 cases. Follow up included 94 patients seen after one week, 63 after 3 months and 10 after 6 months: 3 embolic recurrences were noted (3.2%) of cases. None of the 8 deaths reported was related to the thromboembolic disease. Standard frontal abdominal radiographic images showed migration of filter in 13 cases (13.7%) not exceeding the height of a vertebral body: 9 were caudal and 4 proximal, the LEM* filter remaining within the IVC. Phlebocavography in 90 cases showed the IVC to be permeable in 84 cases (93.3%). Incomplete opening or inclination of filter had no effect on the course. These findings demonstrate that the advantages of the LEM* filter include: a percutaneous introduction allowing rapid, certain insertion, and a form studied for limitation of inclination and avoidance of perforation of the IVC.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3047375 TI - [Systemic artery-pulmonary vein fistula. Apropos of a case]. AB - Fistulas between a systemic artery arising from the aorta and the pulmonary vein with absence of a branch of the pulmonary artery and without abnormality of the bronchial tree are very seldom. We report the case of a child which had a continuous vascular murmur with heart failure secondary to a left-left shunt. A left inferior lobectomy was performed. The inferior lobe was the site of a huge arterio-venous fistula between an artery arising, from the aorta and the pulmonary veins. PMID- 3047376 TI - [Pulmonary hemangiopericytoma. Apropos of a case and review of the literature]. AB - Hemangiopericytoma is a rare vascular tumor of pericyte origin and of variable but always preoccupying potential since falling within the category of sarcomas. A thoracic localization is rare and primary pulmonary parenchyma lesions even more so, only 87 cases being reported in the literature. Radiological diagnostic criteria are of weak specificity, diagnosis being provided by thin section CT scan imaging with contrast. The differential diagnosis of these vascular tumors includes arteriovenous fistula, pulmonary varices, other vascular tumors or even organized bronchomucoceles. The value of CT scan puncture is obviously limited. The course is unpredictable since even histopathology findings fail to provide definitive data. PMID- 3047377 TI - [Echographic and x-ray computed tomographic aspects of intramural hematoma of the duodenum. Apropos of a case]. AB - Radiologic imaging in a case of intramural hematoma of duodenum demonstrated typical appearances of the lesion. Ultrasound imaging of the duodenal hematoma was sufficiently characteristic to be used as a basis for a semeiologic discussion and to orientate diagnosis. The relevant literature is reviewed, and value and indications of different imaging techniques for diagnosis and follow up of the hematoma discussed. PMID- 3047379 TI - Mitchiner memorial lecture, 1987. An example to us all: the military approach to the care of the injured. PMID- 3047380 TI - The efficacy and safety of sulindac (400 mg vs 600 mg daily) in rheumatoid arthritis. A Canadian Multicentre Study. AB - Sulindac, an indene derivative of indomethacin, was compared at 2 dosages (400 mg and 600 mg daily) in patients with rheumatoid arthritis. One hundred sixty-two patients from 19 centers completed the study--85 in the low dose group and 77 in the high dose. No difference in efficacy was found between these 2 regimens. Overall adverse reactions were more frequent with the high dose group especially with respect to blood chemistry and gastrointestinal reactions. PMID- 3047381 TI - Laryngeal infection in lupus: report of nocardiosis and review of laryngeal involvement in lupus. AB - The second reported case of laryngeal nocardiosis in a patient with lupus is described. The manifestations of laryngeal lupus and clinical features that may help to differentiate laryngeal lupus from infection are discussed. PMID- 3047378 TI - [Cecal hernia through Winslow's foramen. Radiographic study of a case and review of the literature]. AB - A case of herniation of the cecum through the foramen of Winslow is reported in a 49-year-old man, admitted for acute abdominal pain. Diagnosis, suspected by abdominal plain films, and established by water-soluble enema, was confirmed by emergency surgical management. Herniation through Winslow's foramen represents the least common variety of internal hernias. Only 136 observations were previously reported, the cecum being involved in 25-30% of cases. Radiological diagnosis was made in only one patient out of ten. Although rare in occurrence, it carries a high mortality risk when diagnosis and treatment are delayed. A better appreciation of the classic radiological features will allow for earlier recognition and treatment. Therefore, the authors emphasize the interest of radiologic findings in this condition. PMID- 3047382 TI - Synovial histology in rheumatoid arthritis: clues for the clinician. PMID- 3047383 TI - NSAID and gastropathy: a rheumatologist's review. AB - Nonsteroidal antiinflammatory drug (NSAID) use, salicylate and nonsalicylate, is all too commonly associated with gastropathy. The problem is reviewed as significantly impacting upon the success of the most common treatment of most forms of arthritis. Blockade of prostaglandin products by NSAID is recently recognized as a basis for the ultimate failure of adaptive cytoprotection to respond to putative threat and restore the gastric mucosa. A cohort of such patients, based upon risk factors especially noted in elderly women taking longterm high sustained doses of NSAID is identified for purposes of closer clinical monitoring. Issues of prophylaxis, adjustment of arthritis regimens to NSAID gastropathy, and gastroprotective measures are reviewed as strategic responses to this recently recognized and described iatrogenic problem of arthritis management. PMID- 3047384 TI - Management of the solitary thyroid nodule. PMID- 3047385 TI - Physical exercise in the prevention and treatment of osteoporosis: a review. PMID- 3047386 TI - Sir Thomas Clarges, apothecary and envoy. PMID- 3047387 TI - Antoni Lesniowski and his contribution to regional enteritis (Crohn's disease). PMID- 3047388 TI - Abdominal aortic aneurysms in identical twins. PMID- 3047389 TI - Disappearance of an Anglechik prosthesis. PMID- 3047390 TI - Vascular compression in thoracic outlet syndrome--a potentially missed diagnosis. PMID- 3047391 TI - Hyperbaric oxygen in multiple sclerosis. PMID- 3047392 TI - Teaching medical students to do bibliographic searching. PMID- 3047393 TI - Use of ultrasound for early pregnancy detection in the rhesus and cynomolgus macaque (Macaca mulatta and Macaca fascicularis). AB - Diagnostic ultrasound provides an accurate method for the detection of early pregnancy and embryonic loss in macaque species. A developing gestational sac (GS) may be observed on gestational day (GD) 14-15 with positive identification on GD 16-18. Visualization of the yolk sac, embryo, and developing heart on GD 21 25 confirms pregnancy. Continuous observations during embryogenesis provide useful information when assessing the teratogenic potential of a variety of agents. PMID- 3047394 TI - Transdermal scopolamine in drooling. AB - The effect of oral anticholinergic drugs has been limited in the treatment of drooling. Transdermal scopolamine (1.5 mg/2.5 cm2) offers advantages. One single application is considered to render a stable serum concentration for 3 days. A distinct reduction of basal salivation was demonstrated in an open trial of six healthy volunteers. Eighteen mentally retarded patients with a drooling problem were studied in a double-blind, placebo-controlled cross-over trial. The therapeutic effect of transdermal scopolamine was assessed by a visual analogue scale. Three patients dropped out due to loss of the system. In the remaining 15 patients, the active drug caused a reduction of drooling which was significant in the period from 24 to 72 h. There were few and slight objective signs of unwanted effects. Scopoderm may cause drowsiness and affect tooth health. The management of drooling should primarily be focused on the cause. Sensomotor training is often valuable in cerebral palsy. Factors such as nasal obstruction, mucosal irritation, and drug-induced parkinsonism should be given attention. Sometimes, however, a temporary symptomatic treatment is indicated, for example on special occasions or in order to cure peri-oral skin lesions. Transdermal scopolamine may offer this possibility. PMID- 3047396 TI - Evaluation of catalytic free energies in genetically modified proteins. AB - A combination of the empirical valence bond method and a free energy perturbation approach is used to simulate the activity of genetically modified enzymes. The simulations reproduce in a semiquantitative way the observed effects of mutations on the activity and binding free energies of trypsin and subtilisin. This suggests that we are approaching a stage of quantitative structure-function correlation of enzymes. The analysis of the calculations points towards the electrostatic energy of the reacting system as the key factor in enzyme catalysis. The changes in the charges of the reacting system and the corresponding changes in "solvation" free energy (generalized here as the interaction between the charges and the given microenvironment) are emphasized. It is argued that a reliable evaluation of these changes might be sufficient for correlating structure and catalysis. The use of free energy perturbation methods and thermodynamic cycles for evaluation of solvation energies and reactivity is discussed, pointing out our early contributions. The apparent elaborated nature of our treatment is clarified, explaining that such a treatment is essential for consistent calculations of chemical reactions in polar environments. The problems associated with seemingly more rigorous quantum mechanical methods are discussed, emphasizing the inconsistency associated with using gas phase charge distributions. The importance of dynamic aspects is examined by evaluating the autocorrelation of the protein "reaction field" on the reacting substrate. It is found that, at least in the present case, dynamic effects are not important. The nature of the catalytic free energy is considered, arguing that the protein provides preoriented dipoles (polarized to stabilize the transition state charge distribution) and small reorganization energy, thus reducing the activation free energy. The corresponding catalytic free energy is related to the folding free energy, which is being invested in aligning the active site dipoles. PMID- 3047395 TI - Spatial arrangement of DNA-dependent RNA polymerase of Escherichia coli and DNA in the specific complex. A neutron small angle scattering study. AB - In this paper we demonstrate that neutron small angle scattering is a suitable method to study the spatial arrangement of large specific protein-DNA complexes. We studied the complex of DNA-dependent RNA polymerase of Escherichia coli and a 130 base-pair DNA fragment containing the strong promoter A1 of bacteriophage T7. Contrast variation of the complex with deuterium allowed us to "visualize" either RNA polymerase, or DNA, or both components in situ. From the corresponding scattering curves information was derived about: (1) Conformational changes of RNA polymerase and DNA by complex formation: comparison of the scattering profiles of the isolated and complexed components showed that by specific complex formation the cross-section of RNA polymerase decreases, while the DNA fragment does not undergo a gross conformational change. (2) The spatial arrangement of RNA polymerase and DNA in the specific complex from the cross-sectional radii of gyration of the complex the normal distance dn between the centre of gravity of the RNA polymerase and the axis of the DNA fragment was derived as 5.0 (+/- 0.3) nm. On the basis of these and footprinting data a low resolution model of the RNA polymerase-promoter complex is proposed. The main feature of this model is the positioning of RNA polymerase to only one side of the DNA. PMID- 3047397 TI - A high resolution diffracting crystal form of the complex between yeast tRNAAsp and aspartyl-tRNA synthetase. AB - Three new crystal forms of the complex between yeast tRNAAsp and aspartyl-tRNA synthetase have been produced. The best crystals, obtained after modifying both purification and crystallization conditions, belong to space group P2(1)2(1)2(1) and diffract to 2.7 A. Unit cell parameters are a = 210.4 A, b = 145.3 A and c = 86.0 A (1 A = 0.1 nm), with one dimeric enzyme and two tRNA molecules in the asymmetric unit. PMID- 3047398 TI - Crystallization and preliminary crystallographic data of a truncated derivative from the human c-Ha-ras protein. AB - A truncated derivative of the human c-Ha-ras protein has been crystallized from polyethylene glycol 6000 solution by a vapour diffusion technique. The rectangular prism diffracts X-rays to at least 2.5 A resolution (1 A = 0.1 nm). The unit cell is monoclinic, space group P21, with unit cell parameters of a = 50.2 A, b = 110.9 A, c = 36.4 A, beta = 97.2 degrees. The unit cell contains four molecules. PMID- 3047399 TI - Intra-chromosomal gene conversion induced by a DNA double-strand break in Saccharomyces cerevisiae. AB - We have stimulated mitotic and meiotic gene conversion between non-tandem direct repeats of ADE4 by a defined double-strand break imparted in vivo to one of two copies of the gene. The experimental design permitted us to distinguish unambiguously between reciprocal intra-chromosomal crossing over and non reciprocal break-join events that could accompany the induced conversions. We observed that (1) less than 10% of the induced conversion events are accompanied by intra-chromosomal crossing over in both mitosis and meiosis; (2) non reciprocal break-join is not stimulated by the double-strand breaks; (3) a double strand break in meiosis is repaired off intra-chromosomal homology (if available) with approximately sevenfold preference over repair off the homologous chromosome. Our observations, analyzed in the light of previous investigations of spontaneous inter and intra-chromosomal crossing over and gene conversion, lead to the view that chromosomal configuration constrains intra-chromosomal crossing over accompanying conversion between closely spaced repeated genes during resolution of the conversion intermediate. PMID- 3047400 TI - Functional sites of the Ada regulatory protein of Escherichia coli. Analysis by amino acid substitutions. AB - Specific cysteine residues at possible methyl acceptor sites of the Ada protein of Escherichia coli were converted to other amino acids by site-directed mutagenesis of the cloned ada gene of E. coli. Ada protein with the cysteine residue at 321 replaced by alanine was capable of accepting the methyl group from the methylphosphotriester but not from O6-methylguanine or O4-methylthymine of alkylated DNA, whereas the protein with alanine at position 69 accepted the methyl group from the methylated bases but not from the methylphosphotriester. These two mutants were used to elucidate the biological significance of repair of the two types of alkylation lesions. Introduction of the ada gene with the Ala69 mutation into an ada- cell rendered the cell more resistant to alkylating agents with respect to both killing and induction of mutations, but the gene with the Ala321 mutation exhibited no such activity. Replacement of the cysteine residue at position 69, but not at position 321, abolished the ability of Ada protein to promote transcription of both ada and alkA genes in vitro. These results are compatible with the idea that methylation of the cysteine residue at position 69 renders Ada protein active as a transcriptional regulator, whilst the cysteine residue at position 321 is responsible for repair of pre-mutagenic and lethal lesions in DNA. The actions of mutant Ada proteins on the ada and alkA promoters in vivo were investigated using an artificially composed gene expression system. When the ada gene with the Ala69 mutation was introduced into the cell, there was little induction of expression of either the ada or the alkA genes, even after treatment with an alkylating agent, in agreement with the data obtained from studies in vitro. With the Ala321 mutation, however, a considerable degree of ada gene expression occurred without adaptive treatment. The latter finding suggests that the cysteine residue at position 321, which is located near the C terminus of the Ada protein, is involved in regulating activity, as the transcriptional activator. PMID- 3047401 TI - Three-dimensional structure of 50 S Escherichia coli ribosomal subunits depleted of proteins L7/L12. AB - A structural study of Escherichia coli 50 S ribosomal subunits depleted selectively of proteins L7/L12 and visualized by low-dose electron microscopy has been carried out by multivariate statistical analysis, classification schemes and the new reconstruction technique from single-exposure, random-conical tilt series. This approach has allowed us to solve the three-dimensional structure of the depleted 50 S subunits at a resolution of 3 nm-1. In addition, two distinct morphological populations of subunits (cores) have been identified in the electron micrographs analyzed and have been separately studied in three dimensions. Depleted subunits in the two morphological states present as main features common to these two structures but different from those of the non depleted subunit (1) the absence of the stalk, (2) a rearrangement of the stalk base that changes the overall structure of this region. This morphological change is quite noticeable and important, since this region is mapped as a part of the GTPase center. The two conformations differ mainly in the orientation of the area between the L1 region and the head (the probable localization of the peptidyl transferase center) and in the accessibility of the region located below the head. A possible relationship of these structural changes to the functional dynamics of the ribosome is suggested. PMID- 3047402 TI - DNA recognition by the FLP recombinase of the yeast 2 mu plasmid. A mutational analysis of the FLP binding site. AB - The 2 mu plasmid of the yeast Saccharomyces cerevisiae encodes a site-specific recombination system consisting of the FLP protein and two inverted recombination sites on the plasmid. The minimal fully functional substrate for in-vitro recombination in this system consists of two FLP protein binding sites separated by an eight base-pair spacer sequence. We have used site-directed mutagenesis to generate every possible mutation (36 in all) within 11 base-pairs of one FLP protein binding site and the base-pair immediately flanking it. The base-pairs within the binding site can be separated into three classes on the basis of these results. Thirty of the 36 sequence changes, including all three at seven different positions (class I) produce a negligible or modest effect on FLP protein-promoted recombination. In particular, most transition mutations are well tolerated in this system. In only one case do all three possible mutations produce large effects (class II). At three positions, clustered near the site at which DNA is cleaved by FLP protein, one of the two possible transversions produces a large effect on recombination, while the other two changes produce modest effects (class III). For seven mutants for which FLP protein binding was measured, a direct correlation between decreases in recombination activity and in binding was observed. Positive effects on the reaction potential of mutant sites are observed when the other FLP binding site in a single recombination site is unaltered or when the second recombination site in a reaction is wild-type. This suggests a functional interaction between FLP binding sites both in cis and in trans. When two mutant recombination sites (each with 1 altered FLP binding site) are recombined, the relative orientation of the mutations (parallel or antiparallel) has no effect on the result. These results provide an extensive substrate catalog to complement future studies in this system. PMID- 3047404 TI - Crystallization and preliminary X-ray investigation reveals that tumor necrosis factor is a compact trimer furnished with 3-fold symmetry. AB - Crystals of recombinant human tumor necrosis factor produced by Escherichia coli have been obtained under different conditions. Crystals suitable for X-ray studies are produced by a vapor diffusion technique using sodium phosphate as both precipitant and buffer at pH 6.5. The crystals belong to the cubic space group, P2(1)3 with unit cell dimensions a = b = c = 95.7 A (1 A = 0.1 nm). Preliminary photography reveals that the crystals are moderately stable to X-rays and diffract to at least 3 A resolution. The diffraction data for native crystals have been collected on a diffractometer at 3 A resolution. Another crystal form, which appeared in a solution containing sodium phosphate at pH 8.0, has the trigonal space group P3 with unit cell dimensions a = b = 63.8 A and c = 54.4 A, and produces measurable reflections to a resolution of 3 A. Hexagonal crystals also have been obtained by the use of polyethylene glycol as precipitant in the range pH 7.6 to 8.0; however, the crystals are fragile and unstable to X-rays. Conservation of 3-fold symmetry in the different crystal forms obtained could reflect the ability of tumor necrosis factor molecules to form trimers in solution and probably the nature of binding of the molecules to cellular receptors. PMID- 3047403 TI - Structure of the rat renin gene. AB - We have isolated the renin gene from a Sprague-Dawley rat genomic library and determined its complete nucleotide sequence. The single rat renin gene is approximately 11,000 bases in length and consists of nine exons and eight introns. The amino acid sequence predicted from the genomic sequence indicates that the rat renin precursor consists of 402 amino acid residues, and shows 85%, 82% and 68% homology to the mouse Ren-1 and Ren-2, and human renins, respectively. The canonical promoter "TATA" boxes, TATAAA and TAATAA, are found 27 and 57 base-pairs upstream from the putative cap site, respectively. Several attractive regulatory sequences analogous to glucocorticoid, estrogen receptor binding sites, cAMP-responsive element and SV40 enhancer core sequences were noted in the 5'-flanking region of the gene. In the first intron, segments with an average size of 38 base-pairs containing a NcoI site are present at 46 tandem repeats within 1710 base-pairs. A "CA" element consisting of (CA)27 was identified in intron 3. Furthermore, intron 8 contains a sequence that shows about 93% homology to that of the neuronal identifier sequence. PMID- 3047405 TI - A look at epikeratophakia. PMID- 3047406 TI - Acquired immunodeficiency syndrome: progress and prospects for vaccine development. PMID- 3047407 TI - Regression models in clinical studies: determining relationships between predictors and response. AB - Multiple regression models are increasingly being applied to clinical studies. Such models are powerful analytic tools that yield valid statistical inferences and make reliable predictions if various assumptions are satisfied. Two types of assumptions made by regression models concern the distribution of the response variable and the nature or shape of the relationship between the predictors and the response. This paper addresses the latter assumption by applying a direct and flexible approach, cubic spline functions, to two widely used models: the logistic regression model for binary responses and the Cox proportional hazards regression model for survival time data. PMID- 3047408 TI - Immunosuppression in the surgical patient. AB - A review of both the healthy and altered immune response as it relates to the surgical patient is presented. An increasing number of immunosuppressive states have been defined in the surgical patient including trauma, burns, sepsis, malnutrition, cancer, and, more recently, acquired immunodeficiency syndrome (AIDS).Investigations of the healthy and altered immune response have led to the development of immunopharmacology, which involves the study of pharmacologic agents that modify host immune response to achieve desired therapeutic goals. Endogenous and exogenous immunomodulators are described that affect uniquely the host immune response as well as their therapeutic implications. PMID- 3047409 TI - Reoperative surgery for persistent hyperparathyroidism. AB - Persistent hyperparathyroidism is an uncommon but recognized sequela of hyperparathyroidectomy that presents difficult diagnostic and surgical challenges. The approach to such patients, particularly in the area of localizing procedures and the conduct of surgery, is reviewed. The successful management of patients with persistent hyperparathyroidism is directly proportional to the surgeon's depth of experience with this entity. PMID- 3047410 TI - Colorectal cancer: surgical management of recurrent and metastatic disease. AB - The surgeon's responsibility to patients with colorectal cancer does not end with resection of the primary locoregional disease. The surgeon has a role to play in (1) designing and implementing strategies aimed at preventing recurrence, (2) early detection of recurrent disease, and (3) resective therapy of recurrent disease in selected instances, either with curative intent or for palliation. To perform these roles, the surgeon must have a thorough knowledge of catheter techniques for regional drug delivery, resective techniques for metastatic or locally recurrent disease, and combined surgical and radiotherapy approaches. PMID- 3047412 TI - AIDS: the role of imaging modalities and infection control policies. AB - The availability of imaging modalities, such as the chest radiograph, gallium scan, double-contrast barium enema, computed tomography, and nuclear magnetic resonance, are very helpful in the diagnosis, treatment, and follow-up evaluation of patients with acquired immunodeficiency syndrome (AIDS). Because this syndrome causes irreversible destruction of the immune system, patients are susceptible to a multitude of opportunistic infections and malignancies. Health care professionals and the general public would be less fearful and apprehensive of AIDS victims if properly informed about the communicability of this syndrome. PMID- 3047411 TI - Update in cancer chemotherapy: genitourinary tract cancer, Part 5: Ovarian cancer. AB - An update of the state of the art of cancer chemotherapeutic treatment of genitourinary tract cancer is described in this multi-part series: included are cancers of the kidney, bladder, prostate, testicle, ovary, uterus, vulva, and gestational trophoblastic neoplasms. Part 5 is a review of treatments for cancer of the ovary.Ovarian cancer is the leading cause of gynecologic cancer deaths and the fourth most frequent cause of death from cancer in women in the United States. Ovarian carcinoma is a classical "hidden" neoplasm of the abdomen and, as such, is insidious in its onset. Combination chemotherapy and cytoreductive surgery, however, have provided improved survival for patients with ovarian cancer. As dosage schedules and other refinements are made, the overall outlook for patients with ovarian cancer is promising. PMID- 3047413 TI - Comparison of the morphological effects of acidic and basic fibroblast growth factors on rat astroblasts in culture. AB - The effects of acidic and basic fibroblast growth factors (aFGF and bFGF) on the morphology of cultured rat astroblasts and on the expression of glial fibrillary acidic protein (GFAP) were compared. The addition of either aFGF or bFGF affected the morphology of the flat, irregular, polygonal-shaped astroblasts, which formed processes and acquire a fibrous appearance. Appreciable different morphological aspects were observed between aFGF- and bFGF-induced cells, essentially between 11 and 14 days in culture. In the presence of bFGF the astroglial cells were more fibrous with a more compact perikaryon as compared to aFGF treated cells. At the ultrastructural level abundant intermediate filaments were observed in astroglial cells as an effect of aFGF and rare filaments but numerous microtubules were seen in bFGF-treated cells. The immunoreactivity for GFAP increased with time in culture and was much stronger in aFGF-treated cells compared to bFGF-treated cells at day 14. An intense positive staining was observed in the somata of the astroglial cells and their processes in the presence of aFGF, while essentially the processes were stained in the presence of bFGF. After 21 days in culture GFAP immunoreaction was also found in the perikarya of cells treated with bFGF. These results show that rat astroglial cells respond somewhat differently to aFGF and bFGF. PMID- 3047415 TI - Pursuing teratogenic causes of multiple congenital contractures. PMID- 3047414 TI - Alterations of the posttranslational processing of a lysosomal enzyme in C6 glioma cells. AB - Cathepsin D was assessed in C6 glioma cells grown in medium with an intermediate- or low-percent composition of serum. The amount, form, and subcellular location of cathepsin D differed after treatment with cyanate or monensin in cells grown in a low-serum, growth-factor-supplemented medium. Immunoblotting showed that cathepsin D in the lysosomal fraction of the C6 cell line had a molecular weight (Mr) of 42 kD, whereas that in the microsomal fraction had Mr's of 42, 47, and 78 kD. After treatment for 1 to 16 hr with 4 mmol/L cyanate and subcellular fractionation, the molecular weight of lysosomal cathepsin D was the same in treated and untreated cells, but more enzyme was found in lysosomes of treated cells at 8 and 16 hr. In the microsomal fraction, the amounts of both the 42 and 47 kD forms were increased after 1 to 16 hr of treatment. When exposed to 20 mmol/L cyanate, C6 cells remained viable, but compared with untreated cells, they showed 25% less lysosomal cathepsin D, with increased amounts found in the microsomal fraction. The 78 kD protein detected by immunoblotting was present in both the lysosomal and microsomal fractions but was predominant in the latter. The apparent molecular weight of this protein was the same after cyanate but differed with monensin, where Mr's of 39, 42, and 73 kD were found. Monensin treated cells had less lysosomal cathepsin D and relatively more microsomal enzyme. The differing molecular weights of cathepsin D from cyanate- and monensin treated cells suggest that their inhibitions occur at different processing loci in distal elements of the Golgi stacks. The differences in the pI of cathepsin D and the number of its forms from cyanate- and monensin-treated cells are also consistent with interference in the late stages of glycoprotein maturation. In this paper we show that the amount, molecular form, and consequent intracellular location of cathepsin D in cells of the C6 line can be affected by agents that selectively disrupt stages in Golgi-related protein modification and transport. PMID- 3047416 TI - Autoimmunity induced by chemicals. AB - Immunotoxicologic studies have demonstrated that autoimmune responses and/or autoimmune diseases are induced in humans and experimental animals by chronic exposure to various chemicals. The present review is focused on seven groups of chemically induced human disorders, i.e. systemic lupus erythematosus, autoimmune hemolytic anemia, myasthenia gravis, pemphigus, glomerulonephritis, thyroiditis and hepatitis. Results obtained from studies of the available experimental counterparts of these diseases, i.e. those models obtained from the exposure of laboratory animals to various chemicals, are then analyzed. Finally, we present the lessons that can be derived from immunotoxicologic investigations regarding mechanisms of induction, heterogeneity of chemicals involved, humoral vs. cellular immune responses and genetic predisposition to chemically induced autoimmunity. PMID- 3047417 TI - Chemical-induced autoimmune reactions and Spanish toxic oil syndrome. Focus on hydantoins and related compounds. AB - Autoimmune diseases comprise a wide spectrum of overlapping, systemic and organ specific disorders. Although, etiology and pathogenesis of such disorders are largely unknown, endogenous host factors and exogenous agents, such as viruses, bacteria, and small molecular weight chemicals, drugs and food components, are believed to be involved. The toxicological significance of low molecular weight compounds on induction of autoimmune disorders is illustrated by the toxic oil syndrome (TOS), a chemically induced epidemic, observed in Spain since 1981. The causative chemical(s) of TOS is still elusive, but an association between ingestion of refined aniline-adulterated rapeseed oil and the syndrome is well documented. Epidemiological, clinical and immunopathological symptoms of TOS are briefly reviewed. The striking resemblance with immunological disorders, observed in man upon medication with hydantoins and related compounds, is demonstrated. The likeliness of formation of a hydantoin-related compound in the aniline adulterated oil is evidenced and its role as possible toxic agent in TOS is proposed. Further, the presence of hydantoins and related compounds in food is briefly reviewed and it is suggested that these chemicals may account for a portion of idiopathic autoimmune diseases observed in man. The need for development of animal models to assess this kind of immunotoxicological effects is stressed. PMID- 3047418 TI - Radio ultrasound observations of the fetotoxic effects in sheep from ingestion of Conium maculatum (poison-hemlock). AB - Fetal movement in pregnant ewes gavaged with Conium maculatum (poison-hemlock) was reduced significantly, but temporarily. Fetal movement was observed by radio ultrasound at 45, 54 and 60 days of gestation in control ewes and on days 45, 54, and 60 of gestation immediately before and 1 hour following poison-hemlock feeding in treated ewes. Fetal movement was significantly reduced (P less than 0.01) 1 hour after poison-hemlock administration, but returned to normal within 18 hours post treatment. At parturition seven of eleven lambs born to seven treated ewes had varying degrees of front limb abnormalities. Modest to moderate flexure of the carpal joints, some lateral deviation in the front limbs at the pastern joint and kinked tails were observed. These malformations were transient and resolved spontaneously by 8 weeks after lambing. PMID- 3047419 TI - Evaluation of coagulation factor abnormalities in long-acting anticoagulant overdose. AB - Newer Rodenticides of the long-acting anticoagulant or "superwarfarin" class are gaining popularity. Since few cases of severe, prolonged anticoagulation after ingestion have been reported, the course of toxicity is not precisely understood. In this case of an intentional ingestion of brodifacoum, a longitudinal analysis of specific coagulation factor derangements was carried out in an attempt to guide a future treatment strategy for this type of toxicity. Results of this analysis demonstrated a profound decrease in levels of factors II, VII, IX, and X, lasting at least 43 days post ingestion. Treatment with subcutaneous vitamin K1 in doses up to 100 milligrams per day was without complication and was effective in reversing the coagulopathy produced by brodifacoum. PMID- 3047420 TI - Prenatal diagnosis of neural tube defects: origin of midtrimester vertebral ossification centers as determined by sonographic water-bath studies. AB - A normal vertebral column was removed from a 17-week stillborn fetus and suspended in a water bath. The specimen was sequentially dissected, with spinous processes, transverse processes, and remaining neural arch removed. After dissection, each specimen was imaged by ultrasound in axial, sagittal, and coronal planes. The resultant images clearly demonstrated the origin of the three ossification centers as one at each lamina-pedicle junction and one in the vertebral body. Corollary X-ray studies confirmed the ultrasound findings and, in a case of open spina bifida, demonstrated the superiority of sonography for making this diagnosis. PMID- 3047421 TI - Neurosonographic changes in newborns treated with extracorporeal membrane oxygenation. AB - Ten out of 40 near-term neonates on extracorporeal membrane oxygenation (ECMO) therapy developed abnormal neurosonograms. We identified two abnormal patterns. Hyperechoic areas of intracranial hemorrhage were observed in two patients (a significantly lower incidence than previously reported). Diffuse or focal echogenic areas of hypoxia-ischemia resulting from periventricular leukomalacia, cerebral edema or large vessel infarction had not previously been noted in nine of these patients. Three of the ten patients survived with neurological sequelae. Recognition of hemorrhage or a hypoxic-ischemic pattern should serve as a warning to initiate or accelerate the weaning of infants from ECMO. PMID- 3047422 TI - Cortical echogenicity in the hemolytic uremic syndrome: clinical correlation. AB - The initial renal sonograms of 15 patients, aged 8 months to 5 years, with hemolytic uremic syndrome (HUS) were reviewed. Ultrasound studies were graded according to cortical echogenicity relative to the liver, they were compared to the severity of the clinical syndrome at admission and to the ultimate outcome of the disease. The degree of cortical echogenicity correlated with the clinical outcome of HUS in 12 of the patients, whereas clinical assessment alone predicted outcome in 13 patients. Sonography overestimated severity in three patients with mild disease correctly assessed clinically, whereas clinical assessment overestimated severity in two patients with moderate disease in whom the sonographic assessment proved correct. The sonographic changes are most likely multifactorial. They appear to reflect a combination of platelet-thrombus deposition in the renal cortex, as well as the general fluid status of the patient. Ultrasound is useful in ruling out other causes of acute renal failure such as obstruction or congenital diseases. It cannot replace laboratory tests and clinical judgement, but nevertheless provides another index of severity in patients with HUS. PMID- 3047423 TI - Ultrasonography of hepatic hydatid cysts: new diagnostic signs. AB - Eighty-five cases of hepatic hydatid cysts were examined by ultrasonography. On the basis of sonographic patterns (as well as clinical symptomatology and surgical, bacteriologic, and pathologic findings), four groups of cysts were identified: uninfected, organizing, suppurative, and degenerated. A specific sign is defined to differentiate hydatid cysts from cystic lesions of other types. Organizing cysts are defined and ways to differentiate them from infected cysts sonographically are discussed. An analysis was made of the echographically detected material between the daughter cysts previously referred to as "matrix." Clues are given to differentiate infected cysts from liver abscesses. A classification correlating the pathology with the life cycle of the parasite in the human body is proposed. PMID- 3047424 TI - Sonography and computed tomography in deep cervical lipomas and lipomatosis of the neck. AB - Nineteen deep cervical lipomas and five patients with cervical lipomatosis were examined with computed tomography (CT) and Sonography. By means of CT, which is the imaging method of choice for both diseases, it is possible to differentiate between circumscribed lipomas and infiltrating intramuscular lipomas. In addition, an exact localization in parenchymatous organs is possible. Cervical lipomatosis is also clearly delineated. Sonography is the first imaging method in cervical swelling or lesions; therefore, knowledge of the sonomorphology of fatty tumors is mandatory. Cervical lipomas have a fairly typical sonomorphology, but it is not as pathognomonic as the density values are by means of CT. In cervical lipomatosis, an adequate pretherapeutic assessment of the depth of infiltration is not possible sonographically. Only the cervical vessels can be clearly differentiated in this condition. PMID- 3047425 TI - Iatrogenic aneurysmal portal-hepatic venous fistula. Diagnosis by color Doppler imaging. PMID- 3047426 TI - Biliary obstruction caused by a papilloma of the common hepatic duct. PMID- 3047428 TI - Testicular feminization syndrome with pelvic seminoma. PMID- 3047427 TI - Ultrasound demonstration of gastroesophageal reflux. PMID- 3047429 TI - Two autonomous myc oncogenes in avian carcinoma virus OK10. AB - The oncogenic avian retrovirus OK10 has the genetic structure gag-delta pol-myc delta-env. The myc sequence is transduced from a cellular gene, proto-myc, while gag, pol, and env are essential retrovirus genes. By analogy with other directly oncogenic retroviruses, the specific myc sequence of OK10 is thought to be essential for transforming function. However, unlike the specific sequences of all other transforming retroviruses that encode unique transforming proteins, the myc sequence of OK10 encodes two potential transforming proteins, p58 and p200. p200 is translated from the gag-delta pol-myc region of genomic RNA, while p58 is thought to be translated from the gag leader and the open reading frame of myc via a subgenomic mRNA. In this paper, we ask whether both myc genes of OK10 are autonomous transforming genes. By differentially inactivating the p200 myc gene of OK10 provirus in vitro and analyzing transforming function in quail embryo cells, it was found that mutants expressing only p58 transformed like wild-type OK10. Further, it was shown that p58 with and without the gag leader had transforming function and that p58 of wild-type OK10 is initiated from the gag leader. Mutants expressing only p200 were also transforming but less efficiently than mutants that express only p58. A mutant OK10 virus in which the native frameshift of retroviruses between gag and pol was deleted expressed a shortened p200 (delta p200). Although this virus expressed more delta p200 than wild-type OK10 did, it transformed cells less efficiently. It follows that OK10 expresses two autonomous transforming genes, which is unique among retroviruses with onc genes. PMID- 3047430 TI - Development of a sensitive quantitative focal assay for human immunodeficiency virus infectivity. AB - Accurate and sensitive quantitation of infectious human immunodeficiency virus (HIV) has been difficult to achieve. In this report, a quantitative focal immunoassay (FIA) for HIV was developed using human HeLa cells rendered susceptible to HIV infection by introduction of the CD4 gene via a retrovirus vector. Infected cells were identified by using human anti-HIV antibodies or mouse monoclonal antibodies specific for HIV together with secondary fluorescein- or peroxidase-conjugated antibody specific for mouse or human immunoglobulins. The assay identified cells infected with either wild-type or culture-adapted HIV isolates and was capable of detecting 1 positive cell in 10(6) cells. The FIA was also effective at detecting cell-free HIV, and in contrast to assays using A3.01, CEM, and other human leukemia cells, the FIA detected most wild-type HIV isolates. HIV neutralization could be determined by using the FIA, and two monoclonal antibodies reactive with HIV gp120 were found to neutralize only the LAV-IIIB strain of HIV. These monoclonal antibodies, as well as antibodies in serum samples from patients with acquired immune deficiency syndrome, were able to inhibit the spread of HIV infection in human lymphocyte suspension cultures but not in CD4-positive HeLa cells growing attached to plastic dishes. PMID- 3047431 TI - Two distinct regions of the murine p53 primary amino acid sequence are implicated in stable complex formation with simian virus 40 T antigen. AB - We mapped regions of the mouse p53 primary amino acid sequence implicated in stable complex formation with simian virus 40 T antigen. A number of mutant p53 proteins failed to complex stably with T antigen in vivo but formed stable complexes with T antigen in in vitro association assays. In contrast to an earlier report (T.-H. Tan, H. Wallis, and A. J. Levine, J. Virol. 59:574-583, 1986), our study showed that two distinct regions of p53 primary amino acid sequence, highly conserved between mouse and Xenopus laevis, were implicated in stable complex formation. Our data support the proposal that, when in complex, T antigen may occupy a site on p53 that is implicated in the normal function of the protein. PMID- 3047433 TI - The prune belly syndrome: a review of its etiology, defects, treatment and prognosis. PMID- 3047434 TI - The natural history of and therapy for perirenal fluid collections following renal transplantation. AB - Fluid collections following renal transplantation are not rare and may be associated with serious complications. We studied the incidence, clinical features, pathology and treatment outcome of perirenal fluid collections after kidney transplantation. Between January 1977 and June 1985, 386 consecutive renal transplants were performed at our university. All allografts were studied with B mode ultrasonography together with a renal scan in the immediate post-transplant period, at 6-month intervals or when clinically indicated. Symptomatic fluid collections, those associated with rejection episodes and those containing more than 50 to 100 ml. fluid were aspirated under sonographic control via aseptic techniques. There were 190 fluid collections (49 per cent) observed during followup (2 to 11 years). Of these collections 98 (51 per cent) were estimated to be less than 50 ml. in volume, were clinically insignificant and resulted in no morbidity. A total of 92 collections was aspirated with 1 aspiration being diagnostic and therapeutic in 57 instances (serous or serosanguinous fluid). The 35 collections remaining were revealed to be lymphoceles on biochemical grounds. Of 13 lymphoceles associated with rejection episodes 8 resolved on initial aspiration. Of the recurrent lymph collections 27 were treated with repeated aspiration, tetracycline sclerotherapy or an operation (10 were treated with marsupialization into the peritoneal cavity). No large collections of urine or blood were detected and 1 infected lymphocele required external drainage. No renal allograft was lost as a result of a fluid collection and over-all graft survival was not affected by the development of perirenal fluid collections. We conclude that perirenal fluid collections are detected commonly in the post transplant period using B-mode ultrasonography. The majority of these collections are small and will require careful observation only or they will resolve with a single aspiration. Aggressive diagnostic and therapeutic measures are used only for those collections that are symptomatic or result in allograft dysfunction. A rational approach to the diagnosis and treatment of peritransplant fluid collections is described in the form of an algorithm. PMID- 3047435 TI - Chronic bacterial prostatitis: 10 years of experience with local antibiotics. AB - Direct transperineal infiltration of the prostate with antibiotics was performed in 24 selected patients with refractory chronic bacterial prostatitis. Diagnosis had been made following a modification of the classical lower tract localization studies. Remission periods of at least 6 months were obtained in 71 per cent of these patients after 1 or 2 infiltrations, while 6 of 24 (25 per cent) required several procedures to assume a long-term remission state and 1 (4 per cent) failed therapy (patient 24490). Seven patients had relapse after remission periods ranging from 13 months to 7 years. The last culture in 3 patients was positive. The value of the method compared to other therapeutic alternatives in the management of chronic bacterial prostatitis is discussed. PMID- 3047432 TI - Passive transfer of respiratory syncytial virus (RSV) antiserum suppresses the immune response to the RSV fusion (F) and large (G) glycoproteins expressed by recombinant vaccinia viruses. AB - In young infants who possess maternally derived respiratory syncytial virus (RSV) antibodies, the antibody response to RSV glycoproteins is relatively poor, despite extensive replication of RSV. In the present study, it was found that cotton rat RSV hyperimmune antiserum suppressed the antibody response to the RSV glycoproteins but not the response to vaccinia virus antigens when the antiserum was passively transferred to cotton rats prior to infection with vaccinia recombinant viruses expressing the RSV envelope glycoproteins. The cotton rats which had their immune responses suppressed by passively transferred antibodies were more susceptible to infection with RSV than were animals inoculated with control serum lacking RSV antibodies. Furthermore, many of the immunosuppressed animals infected with the vaccinia recombinant viruses developed RSV glycoprotein antibodies which had abnormally low neutralizing activities. Thus, preexisting serum RSV antibodies had dramatic quantitative and qualitative effects on the immune response to RSV glycoproteins, which may explain, in part, the poor RSV antibody response of young human infants to infection with RSV. Our observations also suggest that immunosuppression by preexisting, passively acquired RSV antibodies may constitute a major obstacle to RSV immunoprophylaxis during early infancy, when immunization is most needed. PMID- 3047436 TI - Transrectal ultrasonography in the diagnosis and staging of carcinoma of the prostate. AB - Transrectal prostatic ultrasonography is a potentially valuable means to evaluate the prostate of men with suspected carcinoma. We studied 118 patients with this modality before histological evaluation of the prostate (20 underwent radical prostatectomy, 75 core needle biopsy and aspiration cytology, and 23 transurethral resection of the prostate). Transrectal ultrasonography was more efficient than digital rectal examination in the staging of carcinoma of the prostate before radical prostatectomy. The value of transrectal ultrasonography in the diagnosis of prostatic cancer in men with an abnormal-feeling prostate on digital rectal examination is less certain, since 10 of 75 patients (13 per cent) in this group had a falsely positive scan. The predictive value of a scan positive for malignancy was 37 per cent. Further refinements in the technique of transrectal prostatic ultrasonography are needed to realize fully the diagnostic potential of this imaging modality. PMID- 3047437 TI - Flow cytometric determination of ploidy in prostatic adenocarcinoma: a comparison with seminal vesicle involvement and histopathological grading as a predictor of clinical recurrence. AB - Flow cytometry was used to evaluate 88 deparaffinized radical prostatectomy specimens to compare deoxyribonucleic acid ploidy in prostatic adenocarcinoma as a predictor of disease progression with other well documented predictors of clinical recurrence. Aneuploidy, Gleason grade and seminal vesicle involvement were nonindependent variables, and all correlated to statistical significance with disease recurrence. The incidence of aneuploidy in the total population was 51 of 88 (58 per cent). Aneuploidy was found in 25 of 28 primary tumors (89 per cent) from patients who subsequently had recurrent disease. Aneuploidy was noted in 40 of 59 specimens (68 per cent) exhibiting seminal vesicle involvement, compared to 11 of 29 (38 per cent) without seminal vesicle involvement (p less than 0.01). The probability of an interval free of disease of 60 months was calculated by Kaplan-Meier analysis to be 85 per cent in diploid specimens compared to 9 per cent in aneuploid specimens (p less than 0.001), and 87 per cent in the absence of seminal vesicle involvement compared to 28 per cent in the presence of seminal vesicle involvement (p less than 0.003). The probability of remaining free of disease in patients exhibiting concomitant diploidy and seminal vesicle involvement (17) was 73 per cent compared to 8 per cent in those with aneuploid specimens and seminal vesicle involvement (p less than 0.004). The incidence of recurrence in patients with Gleason sums of 7 or less was 22 per cent compared to 62 per cent in patients with Gleason sums of greater than 7. There was a 29 per cent incidence of recurrence in intermediate grade tumors (Gleason sum 5 to 7) but only 5 per cent of the patients with intermediate grade diploid tumors had recurrent disease. In conclusion, the recognition of a deoxyribonucleic acid aneuploid stem line in a primary prostatic adenocarcinoma is correlated statistically with a greater likelihood of seminal vesicle invasion and subsequent development of recurrent disease. The markedly increased probability of remaining free of disease in diploid patients, even in the presence of seminal vesicle involvement, suggests that routine flow cytometric analysis in prostatic adenocarcinoma would significantly enhance prognostic stratification. PMID- 3047438 TI - The extraction factor: an estimate of single kidney function in children during routine radionuclide renography with 99mtechnetium diethylenetriaminepentaacetic acid. AB - An estimate of absolute renal function in children is described using the initial extraction of 99mtechnetium diethylenetriaminepentaacetic acid. The extraction factor correlates well with the glomerular filtration rate as determined by the clearance from the blood of 99mtechnetium diethylenetriaminepentaacetic acid with a correlation coefficient of 0.92. The normal mean extraction factor in the newborn is 1.5 per cent by each kidney, which increases to 2.5 per cent in the first year of life. The method is simple to perform and it is reproducible provided care is taken. We recommend that this procedure be included as part of the routine renal scan in children. PMID- 3047439 TI - Sonographic assessment of normal renal size in children with myelodysplasia. AB - Periodic assessment of upper urinary tract anatomy and renal size is an important component in the urological management of children with myelodysplasia (spina bifida). As a correlation to a previous study of renal growth in children with spina bifida determined by excretory urography, we evaluated 297 renal ultrasonographic examinations in 145 patients with spina bifida and compared the renal size and growth pattern to those of normal children. Patients with known vesicoureteral reflux (grade 2 or greater), congenital renal anomalies, hydronephrosis, renal scarring or urinary tract surgery were excluded. Mean values and standard deviations for sonographically determined renal length were calculated. In general, mean renal length for each age group was below mean values for normal children. A normal renal growth curve for children with spina bifida, based on sonographic renal measurements, is developed for clinical use. PMID- 3047441 TI - The clinical course of carotid bifurcation stenosis as determined by duplex scanning. AB - Two hundred three patients had at least two carotid artery duplex (DS) examinations separated by at least 6 months (mean follow-up 18 months) and their risk factors, clinical outcome, and degree of carotid artery stenosis were compared. The accuracy of DS was established at greater than 90% by blinded comparison with angiography. During the study period 138 patients (68%) had stable carotid artery stenosis and 65 patients (32%) had progression of stenosis in at least one artery. Thirty patients had new symptoms of cerebrovascular disease (19 with transient ischemic attacks and 11 with stroke) during the study. It was not possible to predict which patients would become symptomatic on the basis of risk factor analysis or analysis of severity of stable carotid stenosis. Patients with progression of carotid artery stenosis to greater than 50% diameter reduction were significantly more likely (p less than 0.0001) to have new transient ischemic attack and stroke than were those with stable carotid artery stenosis, regardless of severity. PMID- 3047440 TI - Aortobifemoral bypass: the operation of choice for unilateral iliac occlusion? AB - Aortobifemoral bypass (ABF) is the preferred operation for patients with bilateral aortoiliac occlusive disease, but for those with unilateral occlusion without significant stenosis of the contralateral iliac artery, alternative reconstructions, such as femorofemoral (FF) or iliofemoral (IF) bypass have been advocated. We compared the surgical outcome in 96 such patients after ABF (n = 32), FF (n = 47), or IF (n = 17) bypasses, with biplane arteriography and noninvasive laboratory testing used to assess the contralateral iliac artery and runoff status, in particular, patency of the superficial femoral artery (SFA). Graft patencies were assessed by noninvasive criteria and analyzed by the life table method. The only death occurred after ABF bypass (3.1%). Primary patency rates at 1, 3, and 5 years with an open SFA were 100%, 89% and 89%, respectively, for ABF; 92%, 92%, and 92% for FF; and 71%, 71%, and 36% for IF. When the SFA was occluded, the primary patency rates at 1, 3, and 5 years were 100%, 100%, and 72%, respectively, for ABF; 72%, 53%, and 35% for FF; and 56%, 56%, and 56% for IF bypasses. There were no later occlusions on the contralateral ("good") side after ABF. Significant progression of atherosclerosis in donor iliac artery was observed in 6% of both FF and IF bypasses. We conclude that ABF is the preferred operation for extensive iliac artery occlusive disease that is hemodynamically significant only on the symptomatic side unless specifically contraindicated by prohibitive risk or abdominal disease. This is particularly true in the face of SFA occlusion. PMID- 3047443 TI - Perioperative and late outcome in patients with left ventricular ejection fraction of 35% or less who require major vascular surgery. AB - Survival in patients with diminished left ventricular ejection fraction (EF) is reduced after major vascular surgery. The objective of this study was to determine perioperative (30-day) and subsequent outcome after major vascular surgery in those with severe cardiac dysfunction, defined by EF being 35% or less (normal EF greater than 50%). From Aug. 1, 1984 to Jan. 1, 1988, 35 patients with EF equal to 27.7% +/- 6.1% (mean +/- 1 standard deviation) have required 47 major vascular procedures: 53% (n = 25) were limb revascularizations; 21% (n = 10) were direct aortoiliac aneurysm repairs: 23% (n = 11) were carotid endarterectomies: one patient had transaortic renal endarterectomy. Two deaths occurred within the first 30 days, yielding a 4.3% perioperative mortality rate (2 of 47 procedures). The cumulative mortality rate for the entire group during follow-up of 410 +/- 390 days was 40% (14 of 35 patients). Most late deaths (71%) occurred within the first 6 months after surgery and each was due to cardiovascular complications. Survival for those with an EF of 29% or less was significantly worse than for those with an EF greater than 29%, determined by life-table analysis (p less than 0.012, Mantel-Cox). The cumulative mortality rate was 59% with an EF of 29% or less and 18% in those with an EF greater than 29% (p less than 0.029, two-tailed Fisher exact test). The perioperative mortality rate for those with an EF of 35% or less who require major vascular surgery is acceptable, but overall survival during follow-up is diminished.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3047442 TI - Duplex ultrasound assessment of venous diameters, peak velocities, and flow patterns. AB - Duplex ultrasound was used to study the diameters, flow patterns, and peak blood flow velocities of the common femoral vein (CFV) in 12 normal subjects (mean age 35 years). Each subject was supine and non-weight-bearing on a tilt table and rotated in 10-degree increments from -10 degrees (head down) to +30 degrees (head up). Cross-sectional B-mode image was used to monitor continuously CFV diameter for 5 minutes in each position. Doppler flow patterns were recorded in longitudinal axis; heart rate and respiratory movements were also noted. CFV flow was affected by respiratory and cardiac events. At -10 degrees flow was primarily related to cardiac events, with flow increasing during diastole. At +30 degrees flow varied minimally with the cardiac cycle and was primarily respiration dependent, stopping at peak inspiration. Proceeding from -10 to +30 degrees the mean maximal CFV diameter corrected for body surface area increased 92% (0.47 +/- 0.11 cm/m2 to 0.90 +/- 0.16 cm/m2, p less than 0.001), whereas peak flow velocity decreased from 41 +/- 10 cm/sec to 13 +/- 5 cm/sec, p less than 0.001. There was a linear, inverse relationship between mean peak velocity and mean corrected diameter, r = -0.99. The study confirms the multiple influences on venous flow patterns and establishes a quantitative relationship between venous diameters and flow velocities. PMID- 3047444 TI - Routine revascularization with resection of infection femoral pseudoaneurysms from substance abuse. AB - Infected femoral artery pseudoaneurysms in narcotic addicts present challenging management options. Our policy of routine revascularization is based on the concern that a high rate of amputations must follow ligation and resection alone or with selective delayed revascularization. Fifteen of 16 patients with infected pseudoaneurysms of femoral arteries, treated with resection and bypass grafts, were observed from 1 to 44 months. Obturator bypass grafts were used in 10 patients, iliac-femoral grafts in three, axillopopliteal in one, and right external iliac crossover to left popliteal in one patient. One limb, unsalvageable at presentation, was amputated primarily, along with resection of pseudoaneurysm and femoral artery ligation, without bypass grafting. One iliac femoral graft became infected and then thrombosed 4 months after operation. Unsuitable distal arteries and impending necrosis led to above-knee amputation. One late failure among 15 revascularization attempts (7%) is significantly lower than the 11% to 33% amputation rates reported in the literature with resection of pseudoaneurysm alone and delayed selective revascularization. The other 14 patients had functioning limbs without claudication or rest pain. Our experience indicates that revascularization at the time of resection of infected pseudoaneurysm offers better prospects for limb salvage. PMID- 3047445 TI - Iatrogenic superior mesenteric arteriovenous fistula. Report of a case and review of the literature. AB - Bowel resection can result in the formation of an arteriovenous fistula at the point of intersection of the feeding vessels. Iatrogenic arteriovenous fistula of the superior mesenteric vessels is rare, with only 11 cases reported in the English language literature. We report a case after small bowel resection, which was done because of adhesions from a laparoscopic sterilization. An anatomic classification of the fistulas into H and U types is suggested, with its significance for management. A review of the literature is included. PMID- 3047447 TI - HIV-1 antibody testing. PMID- 3047446 TI - Aortobifemoral perigraft abscess: treatment by percutaneous catheter drainage. AB - There is significant morbidity and mortality associated with infected prosthetic aortic grafts. The preferred method of treatment is excision of the involved graft and revascularization. Percutaneous catheter drainage of abscesses under CT scanning or ultrasound guidance has been shown to be an effective alternative to surgery. This is a case of a patient with a retroperitoneal abscess involving the femoral limb of an aortobifemoral bypass graft, which was successfully treated by percutaneous catheter drainage. In selected high-risk patients this may be a safe alternative to surgical therapy. PMID- 3047449 TI - The epidemiology of acquired immunodeficiency syndrome among heterosexuals. PMID- 3047448 TI - Effects of prophylactic lidocaine in suspected acute myocardial infarction. An overview of results from the randomized, controlled trials. AB - The effects of prophylactic lidocaine hydrochloride on early ventricular fibrillation and death in patients with suspected acute myocardial infarction were investigated in an overview of 14 randomized trials. During follow-up intervals of one to four hours in the trials of intramuscular lidocaine infusion (6961 patients) and 24 to 48 hours in the trials of intravenous lidocaine injection (2194 patients), a total of 103 cases of ventricular fibrillation and 137 deaths were recorded. Overall, allocation to lidocaine was associated with a reduction in the odds of ventricular fibrillation of about one third, with a 95% confidence interval that ranged from a 3% to a 56% reduction. There was no evidence of any beneficial effect on early mortality; indeed, the odds of early death were about one third greater among patients allocated lidocaine, though this difference was not statistically significant (95% confidence interval, 2% reduction to 95% increase). Because of the small numbers of reported events, the short follow-up periods, and the unavailability of data for some specific causes of death, even an overview of all the trial results does not provide good evidence as to whether prophylactic lidocaine is likely to be helpful or harmful. To answer this question reliably, future trials will need to involve large numbers of patients and prolonged follow-up. PMID- 3047450 TI - Obituary. Hans Popper, MD, PhD. PMID- 3047451 TI - Gonadal activity and chemotherapy-induced gonadal damage. PMID- 3047452 TI - A current look at the rationale of the Institute of Medicine of the National Academy of Sciences. PMID- 3047453 TI - Commentary by the current president of the Institute of Medicine. PMID- 3047454 TI - Still needed: a National Academy of Medicine. PMID- 3047455 TI - Neurological complications of central venous cannulation. AB - The reports of neurological damage after central venous cannulation over the past 20 yrs have been gathered, summarized, and analyzed. We found 59 cases of nerve lesions: 32 serious or even fatal, and 27 light or transient ones. They included: Lesions of the cervical sympathetic chain: needle trauma, compression by hematoma, anesthetic blockade; brachial plexopathies: needle trauma or compression by hematoma; phrenic or recurrent nerve palsies: anesthetic blockade, needle trauma, or compression by hematoma; cerebral damage following venous air embolism, carotid artery embolism or obstruction (thrombosis, compression), or internal jugular vein obstruction (thrombosis or catheter tip); lesions of the IX, X, XI, and XII cranial nerves by hematoma compression or spilling of histotoxic solutions; and, massive lesions of the anterior rami of the cervical nerves by spillage of histotoxic solutions. We believe that the following simple and well-known measures can substantially reduce the incidence of those serious complications. 1. Avoiding the subclavian or jugular central routes in patients with marked anatomical changes, coagulopathies, and carotid artery or lung diseases. 2. Using only small amounts of dilute concentrations of short-acting local anesthetics before the puncture. 3. Using a small gauge "seeking" needle and placing a finger on the carotid artery during a jugular venipuncture to avoid accidental arterial puncture. 4. Using radiopaque catheters long enough to have their tip in midsuperior vena cava. The position of the catheter must be checked radiographically immediately after insertion and even at later periods. The catheter must be meticulously fixed to the skin to avoid its movement.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3047457 TI - [General remarks on glomerulonephritis. Classification of glomerulonephritis and historical changes]. PMID- 3047456 TI - [Acute pericarditis caused by daunorubicin in acute myelocytic leukemia]. PMID- 3047459 TI - [Various types of glomerulonephritis. Glomerulonephritis following kidney transplantation]. PMID- 3047458 TI - [Various types of glomerulonephritis. Mesangium proliferative glomerulonephritis- IgA nephropathy]. PMID- 3047460 TI - [Glomerular diseases secondary to diabetes mellitus]. PMID- 3047461 TI - [Clinical usefulness of stereoscopic digital subtraction angiography]. PMID- 3047462 TI - [Iodine scintigraphy of differentiated thyroid carcinoma]. PMID- 3047463 TI - [A case of cavernous hemangioma with large hematoma]. PMID- 3047464 TI - [The 7 cases of cholelithiasis in infant]. PMID- 3047466 TI - [Renal disorders induced by organic solvents]. AB - Organic solvents are a class of chemical substances that are widely used in large volume in various manufacturing plants. Due to their high lipophilicity, organic solvents may produce physicochemical damage to renal glomeruli and tubuli. Many reports have been published abroad, though only a few in Japan, on organic solvent-induced nephropathy such as Goodpasture syndrome, a form of renal damage mediated by anti-basement membrane antibodies. This article reviews the epidemiological studies published in the literature and stresses that organic solvent exposure is one of the etiological factors involved in the development of chronic nephropathy. At present in Japan there are 60,000 patients with terminal renal failure undergoing hemodialysis, 50,000 young men placed under observation each year for abuse of organic solvents, and one million workers exposed to organic solvents in factories. Attention should be focused on the occurrence of renal damage due to organic solvents in Japan. PMID- 3047465 TI - [A case of lupoid hepatitis associated with glomerulonephritis--concerning the findings of renal lesion by immunofluorescence microscopy]. PMID- 3047467 TI - [Thallium-201 myocardial single photon emission computed tomography after isoproterenol infusion in diagnosing ischemic heart disease]. PMID- 3047468 TI - [Fundamental and clinical study of direct immunoradiometric assay of human renin concentration]. PMID- 3047469 TI - Prevalence of fatty liver in a general population of Okinawa, Japan. AB - A total of 2,574 residents in Yaeyama District of Okinawa, Japan, were investigated using real time ultrasonography to determine the real prevalence of fatty liver in the general population and to define its associated factors. Overall prevalence of fatty liver was 14.0%. Prevalence of fatty liver in persons under 19 years old was only 1.2%, and increased with age to a maximum in persons 40-49 years of age and then decreased. For persons over 20 years old, obesity index and serum levels of triglyceride and total cholesterol were measured, and alcohol consumption was asked. Prevalence of fatty liver was significantly higher in drinkers than non-drinkers (p less than 0.01), and increased with alcohol consumption. Furthermore, in persons not suffering from obesity prevalence of fatty liver was significantly higher in drinkers than in non-drinkers (p less than 0.001). The results of logistic regression analysis indicated that obesity and elevated serum triglyceride level in both sexes, and alcohol in males were significant predictors of fatty liver. In conclusion, prevalence of fatty liver increased with age to a maximum in persons 40-49 years of age and overall was 14.0%. Obesity was the strongest associated factor in both sexes and in males alcohol was also a strong factor. PMID- 3047470 TI - Hypoglycemia in the elderly. AB - Fifty three elderly hypoglycemic patients seen at Tokyo Metropolitan Geriatric Hospital during last 4 years were reviewed, and these data suggest that clinical characteristics of hypoglycemic patients in the elderly are as follows: 1) Non diabetics without any hypoglycemic drugs seem to take relatively large part in hypoglycemic patients; 2) malnutrition due to chronic disease, and liver dysfunction may be important predisposing factors in development of hypoglycemia; 3) they are often lacking in adrenergic symptoms and do not tend to present symptoms until their plasma glucose levels decrease at extremely low levels, moreover in such cases, they tend to manifest neuroglycopenia in the beginning of their hypoglycemic attacks. Therefore, it is noteworthy that impaired subjective recognition may increase the risk of hypoglycemia and hypoglycemia should be one of the important considerations in differential diagnosis of stupor and coma in the elderly patients, even though they have not been administered any hypoglycemic agents. PMID- 3047471 TI - [History of nursing in Japan. II. The modern era (3) Nursing activities during the Sino-Japanese war of 1894-5 and the Russo-Japanese war; (4) Prevalence of the nursing system and legislation of nursing regulation]. PMID- 3047472 TI - [Survey on planning of clinical training in nursing education]. PMID- 3047473 TI - [History of nursing in Japan. 12. The modern era. 4. The nursing system and education during the Taisho era (1). The life and health of the citizen during the era (2). The First World War and nursing activities during the Siberian campaign (3). Implementation of the nursing regulations by the Ministry of the Interior. (4) Start of college level nursing education]. PMID- 3047475 TI - [Echography in the topical diagnosis of metastases of malignant neoplasms to the cervical lymph nodes]. PMID- 3047474 TI - [A case report of mediastinal Castleman lymphoma associated with superior vena caval syndrome]. PMID- 3047476 TI - [Prof. Vera Ignat'evna Gedroits, the first woman-surgeon in Russia]. PMID- 3047478 TI - [Advances and perspectives in kidney transplantation]. PMID- 3047477 TI - [Intraoperative ultrasonic examination in abdominal surgery]. PMID- 3047479 TI - [Stomach burns (review of the literature)]. PMID- 3047480 TI - [Scientific surgical school of Academician B.V. Petrovskii (on his 80th birthday)]. PMID- 3047481 TI - [Vascular microsurgery in cardiology]. PMID- 3047482 TI - [Simultaneous total colectomy with ileorectostomy in severe nonspecific ulcerative colitis in children]. PMID- 3047483 TI - [Surgical treatment of patients with an extreme degree of alimentary constitutional obesity]. PMID- 3047484 TI - [The role of immune mechanisms in the etiology and pathogenesis of peptic ulcer]. PMID- 3047486 TI - [The historical medicine significance of eponymous clinical terms]. PMID- 3047487 TI - [Social psychological aspects of the study of medical problems in memoir sources]. PMID- 3047485 TI - [The Museum of Medicine of the Ukrainian SSR]. PMID- 3047488 TI - [The Advanced Therapy Clinic of the Sechenov 1st Moscow Medical Institute in 1917 1987]. PMID- 3047489 TI - [Immunology problems of peptic ulcer]. PMID- 3047490 TI - [Local factors in the pathogenesis of peptic ulcer]. PMID- 3047491 TI - [New possibilities in the treatment of cholelithiasis]. PMID- 3047492 TI - [Mirizzi's syndrome]. PMID- 3047493 TI - [Treatment of bronchial asthma in middle-aged patients]. PMID- 3047494 TI - [Problems of variability in weightlessness]. AB - The genetic (cytogenetic) effects of microgravity have been under study for about 30 years. This line of research developed through three periods. Initially, biologists raised the question whether microgravity may act as a lethal factor or a strong mutagen. Many flight experiments gave a negative answer to it. The major goal of the second period was to identify the effects of microgravity at the chromosomal and cellular levels. Such effects, statistically significant although small in value, were detected in Tradescantia paludosa and Drosophila melanogaster microspores. These findings were confirmed by American and--later- by European investigators. The third period addresses the entire problem of variability in microgravity. This approach can be implemented on the basis of a comprehensive program of research that will include experiments at different levels: from the gene to the population level. The basic objective of the program is to clarify the role of the genetic apparatus in the adaptation of living systems to microgravity and, consequently, to determine the role of gravity in the evolution of life on the Earth. PMID- 3047495 TI - [Energy reactions in the skeletal muscles of rats after short-term space flight on Kosmos-1514]. AB - Ten hours after the 5-day space flight on Cosmos-1514 rats were examined for oxidative phosphorylation in mitochondria isolated from the posterior femoral muscles as well as for Krebs cycle enzymes and glycolysis in the mitochondrial and cytoplasmic fractions of the muscles. The mitochondrial respiration rate in various metabolic states was similar in flight rats and vivarium controls. After flight calculated parameters of energy efficacy of respiration as well as activity of malate dehydrogenase, isocitrate dehydrogenase and total lactate dehydrogenase remained unchanged. Unlike the flight rats, the synchronous controls showed signs of the stress-reaction: uncoupling of oxidative phosphorylation and oxalacetate inhibition of succinate dehydrogenase. Comparison of these findings with those from prolonged space flights indicates that inhibition of oxidative metabolism and glycolysis in mixed muscles which was demonstrated in the 20-day space flight does not develop immediately after launch but occurs within the time interval between mission days 6 and 18. PMID- 3047496 TI - [Comparative study of decompression-induced formation of gas bubbles using ultrasonic equipment and the development of altitude-decompression disorders]. AB - 442 altitude experiments on 40 volunteers were performed. Gas bubbles in venous blood were detected using an ultrasonic Doppler system functioning in a continuous mode at a frequency of 5 MHz. The threshold of bubble formation was identified in 31 test subjects and that of emergency of altitude-decompression disorders, in 28 test subjects. Individual resistance to decompression-induced bubbles was revealed. It was shown that the thresholds may vary with time, in the head-down position and in the head-down position combined with exercises. A correlation was established between the rate of intravascular bubble formation and the frequency of altitude-decompression disorders. PMID- 3047497 TI - Methods for studying protein synthesis and degradation in liver and skeletal muscle. AB - Different methods used for measuring protein turnover in liver and skeletal muscle are described, with special emphasis on technical and practical aspects and the advantages and limitations of different techniques. In the first part of the review, the concept of precursor specific radioactivity and its importance for accurate determination of protein synthesis rate is discussed. In the second part, different in vivo techniques for protein turnover measurements are reviewed, including continuous administration of tracer amino acid, flooding dose technique, indirect measurement of protein synthesis, and estimation of protein degradation in vivo. In the third part of the report, in vitro techniques are described, including measurement of protein turnover in incubated liver slices, perfused liver, isolated hepatocytes, incubated isolated muscles or muscle biopsies, and perfused rat hemicorpus. In vivo techniques are preferred when accurate absolute values of protein turnover rates are desired. In vitro techniques offer the advantage of standardized conditions, maintaining strict control of substrate and hormone concentrations, and eliminating complicating interactions with other tissues. For several in vitro techniques, a good correlation has been demonstrated between relative changes in protein turnover in vitro and in vivo in different conditions. PMID- 3047499 TI - Primary malignant lymphoma of the mandible. AB - From 1945 through 1985, 32 cases of primary lymphoma of bone were treated at the University of Iowa Hospitals and Clinics. Sixteen cases (50%) demonstrated the lesion in the long tubular bones with a predilection for the lower and upper extremities. The frequently involved flat bones (six cases) were the bones of the pelvis. There were only three cases (9%) where the mandible was the primary site. In this report, the literature is reviewed and three cases with primary lymphoma of the mandible are presented. PMID- 3047498 TI - Skin cancer of the nose: options for reconstruction. AB - The nose is the most common site of skin cancer on the human body. This review was compiled to enumerate the many reconstructive techniques which are available and to discuss the advantages and disadvantages of each technique. Five hundred twenty-one patients with nasal cancer were treated at Roswell Park Memorial Institute from 1964 to 1981. Four hundred five patients were treated by cryotherapy, electrocautery, chemosurgery, radiation therapy, or local excision with primary closure. Forty-six patients were treated with simple nasolabial flaps. The remaining 70 patients underwent more extensive resections and reconstructions. Their records were reviewed. At 5 years the local recurrence rate was 3% for basal cell carcinoma and 20% for squamous cell carcinoma. The multiple reconstructive techniques which were utilized in these patients are discussed. Skin-grafting procedures were used to manage 51% of the patients. Prostheses were used in 9%. Multiple different reconstructive flaps were used for the remaining 40% of the patients. The management of large nasal carcinomas must be individualized with therapeutic decisions being made by a practitioner who is familiar with the many reconstructive options. This will ensure the optimal functional and aesthetic result for each patient. PMID- 3047500 TI - Testicular cancer in young Norwegians. AB - The clinical experience is reviewed in 597 Norwegian testicular cancer patients (age range: 15-45 years) treated from 1979 to 1986. During this period, computer tomography, determination of serum AFP/HCG, and cisplatin-based chemotherapy represented the modern diagnostic and therapeutic modalities. Before orchiectomy 67% of the patients had elevated AFP/HCG. An abnormal postorchiectomy serum tumour marker decrease and the presence of small vessel infiltration in the histological sections of the primary tumour significantly predicted microscopic retroperitoneal metastases in patients with clinical stage I (CSI) nonseminoma. One-third of these patients had a pathological stage II (PSII). After radiotherapy 99% of 90 seminoma patients (CSI/IIa) survived for 5 years. After cisplatin-based chemotherapy (+radiotherapy/surgery) the 5-year survival rate in 25 patients with advanced seminoma was 81%. The survival rate in 148 nonseminoma patients PSI/IIa was 100% and 87% in 94 patients with advanced nonseminoma (greater than or equal to CSIIb). Nausea, general exhaustion, myelosuppression, peripheral neuropathy, and Raynaud-like phenomena were the main acute treatment related side effects. Slight gastrointestinal problems, slight peripheral neuropathy, Raynaud-like phenomena, and fertility disturbances were frequent late side effects. The sexual life in testicular cancer patients did not seem to be significantly impaired as compared to the normal population. Most of the patients reported no or only slight emotional problems during and after treatment. The need of thorough information at the time of diagnosis was stressed by most of them. Secondary cancer was diagnosed in 27 of 795 patients (1970-1982) (Testicular: 15; pulmonary: 4; sarcoma: 2; others: 6). Testicular cancer is today a curable malignancy. Future clinical research has to concentrate on the identification of high-risk and low-risk patients, the avoidance of overtreatment, and the reduction of toxicity (especially of long-term side effects). PMID- 3047501 TI - Renal angiomyolipomas. AB - Renal angiomyolipomas are generally benign tumors which are composed of smooth muscle, fat, and blood vessels. Classically these have been diagnosed with the use of arteriograms; however, since ultrasound and CAT scans have come into use these now are the common modalities for diagnosis. All angiomyolipomas generally can be followed conservatively, whereas larger ones have a tendency to bleed and require intervention. This paper reviews the diagnosis, treatment, and prognosis of patients with angiomyolipomas. PMID- 3047502 TI - Malignant epithelioid schwannoma: a light microscopic and immunohistochemical study. AB - Malignant epithelioid schwannoma is a rare variant of malignant nerve sheath tumor that can be confused with both other neuroectodermal neoplasms and carcinomas. The light microscopic and immunohistochemical findings of a malignant epithelioid schwannoma arising in the mandibular region of a 27-year-old female are described. The differential diagnosis of this unusual neoplasm from malignant melanoma and poorly differentiated carcinoma is discussed. PMID- 3047503 TI - [Steiner and nursing education]. PMID- 3047504 TI - Older veterans' future use of VA health care services. AB - This study is a secondary analysis of the Harris Survey of Aging Veterans (SAV) and is designed to identify variables that may be associated with older veterans' future use of the Veterans Administration (VA) health care system. Using regression and discriminant analysis techniques, the study identifies variables that may predispose older veterans to use the VA health care system in the next 10 years. The results indicate that older veterans may elect to use their health care benefits on objective criteria consistent with their health and financial resources, e.g., past use of veterans benefits, expected health status, and private insurance coverage. These variables suggest that the VA's recently enacted means test and the removal of automatic age eligibility will disenfranchise few older veterans. PMID- 3047505 TI - Work capacity of older men and age-eligibility for Medicare benefits. AB - Whether the functional capacities of older Americans enable them to remain in the work force longer, is a significant consideration in appraising proposals to advance the normal eligibility age for Medicare benefits. This article analyzes the durations of work capability and work disablement of a representative panel of 5000 white and black men in their sixties and early seventies. The analysis shows that these men and succeeding cohorts of men are expected to function capably for long enough periods to meet the conditions necessary for raising the age of Medicare eligibility. PMID- 3047506 TI - [Expectations in the increase of survival of patients with cardiac insufficiency treated with angiotensin-converting enzyme inhibitors]. PMID- 3047507 TI - [Hemorrhage caused by peptic ulcer: are there criteria for a more logical treatment?]. PMID- 3047508 TI - [Familial chondrocalcinosis]. PMID- 3047509 TI - Antiplasmid activity of tricyclic compounds. AB - Three representative groups of tricyclic psychopharmacons were shown to have antiplasmid activity. The first group, e.g., comprises promazine, chlorpromazine, promethazine, levo- and dextromepromazine, thioridazine, diethazine, thiethylperazine. There are two heteroatoms (S and N) in the ring system of these compounds. In the second group, which includes desipramine, imipramine, however, there is only one heteroatom N, or S in chlorprothixene, chlorpenthixols and in the third group amitriptyline, maprotiline contain no heteroatoms. The medical significance of plasmid elimination by tricyclic psychopharmacons and related compounds in vitro is that this opens up a new perspective in the struggle against bacterial resistance to antibiotics. This approach utilizes the beneficial side-effect of the plasmid curing ability of these psychopharmacons, and may serve as a guideline for drug design in the future, to develop related substances which are potent in eliminating the drug resistance-carrying plasmids of bacteria, but which have no other pharmacological effects. Here we propose a new mechanism for plasmid curing by tricyclic drugs. PMID- 3047510 TI - [Investing in experiments and independent organizations. You can create the world's best information system]. PMID- 3047511 TI - [Homeopathy in the dog-days--how to refresh our memory?]. PMID- 3047512 TI - [Cancer in man caused by virus--an epidemiological overview]. PMID- 3047514 TI - 200 years of nursing. A photographic history. PMID- 3047513 TI - [ISIS-2--a study of patients with myocardial infarction. A combination of streptokinase and acetylsalicylic acid diminishes mortality risk, reinfarction and stroke]. PMID- 3047516 TI - [Radioprotective action of mexamine in fractionated irradiation]. PMID- 3047517 TI - [Quality guarantees in radiation therapy (a review of the literature)]. PMID- 3047515 TI - Dexamethasone increases the capacity of urea synthesis time dependently and reduces the body weight of rats. AB - Rats of 230 g were treated with 0.1 mg of dexamethasone twice daily for 2 days (n = 5) and 14 days (n = 9). Controls received isotonic saline. During the first week of dexamethasone treatment the rats lost weight rapidly (up to 9 g/day). The weight loss diminished during the second week of treatment. The fasting blood insulin concentration increased sevenfold in the dexamethasone-treated rats. Fasting blood glucagon and glucose concentrations were not different from controls. In the dexamethasone-treated rats the fasting alpha-amino-N concentrations were lower: 4.0 +/- 0.3 mmol/l (mean +/- SEM) versus 6.8 +/- 0.3 mmol/l in controls. The capacity of Urea-N Synthesis, determined during alanine loading was: after 2 days of treatment 14.7 +/- 1.7 mumol/(min 100 g), after 14 days of treatment 7.9 +/- 0.8 mumol/(min 100 g), and in controls 7.5 +/- 1.0 mumol/(min 100 g) (mean +/- SEM). In conclusion, glucocorticoid treatment leads to a transient change in the liver function as to hepatic amino-N conversion, implying that more amino-N than normal is eliminated as urea-N after 2 days of treatment. This may contribute to the early, but not the late body weight loss. PMID- 3047518 TI - [Luminophores on a base of rare-earth element compounds for X-ray intensifying screens]. AB - Presents the results of measuring the traits of intensifying X-ray screens, made with the use of following luminescent solids: Y2O2S:Tb (P-14-1, P-14-2), BaFCI:Eu (P-15-1), CaWO4 (P-420, P-420-1). Summurizes the experience in the application of the screens EU-I1, EU-I5 and EU-IB1 in the All-Union Scientific Center on Surgery, the USSR Academy of Medical Sciences. PMID- 3047519 TI - The effect of glipizide on extraction of insulin by the human cirrhotic and noncirrhotic liver. AB - The effect of glipizide on hepatic uptake of insulin was studied in five patients with liver cirrhosis and five patients with varying diseases in the biliary system and pancreas. All patients had portal catheters for diagnostic purposes. The hepatic uptake of insulin was estimated from the clearance rate for insulin obtained after a constant rate infusion into a peripheral vein and the portal vein. Each patient was examined on two consecutive mornings, the second investigation carried out one hour after oral glipizide administration. The fractional hepatic uptake was significantly lower in cirrhotic patients (17% +/- 6%) than in the other patients (52% +/- 7%; P less than .01). After glipizide, an increase in the estimated uptake of insulin occurred in cirrhotic patients (from 17% +/- 6% to 39% +/- 6%; P less than .01), whereas an insignificant decrease was observed in the other patients (from 52% +/- 7% to 43% +/- 11%). PMID- 3047520 TI - Improved glycemic control lowers plasma apoprotein E and triglyceride levels following ingestion of a fat load in insulin-dependent diabetic subjects. AB - To assess the effect of glycemic control on triglyceride (TG) and apoprotein E (apo E) metabolism, plasma levels of TG and apo E were studied in nine nonobese subjects with insulin-dependent diabetes mellitus (IDDM) following acute ingestion of polyunsaturated fat. Each subject was studied twice: before and after ten days of continuous subcutaneous insulin infusion (CSII). Each subject ingested identical meals on both study days. Plasma glucose was determined in all patients before and two hours after each meal and at 3 AM, and a mean value was calculated for each patient. CSII reduced mean plasma glucose from 205 to 113 mg/dL (P less than .005, paired t test); there was no change in the total daily insulin dose. Plasma TG and apo E levels were measured before and 3.5, 5, and 7 hours after a breakfast which contained 50 g of fish oil (five subjects) or vegetable oil (four subjects). A repeated-measures ANOVA was performed to assess the effects of the following three factors on plasma TG and apoE levels: type of oil ingested (Oil, factor A), glycemic control (Glycemic control, factor B), and the response to fat ingestion over time (Times, factor C). Plasma levels of both apo E and TG increased significantly after fat ingestion (F test, ANOVA, P less than .005 and P less than .001, respectively). Glycemic control significantly reduced the rise in both apo E and TG levels (P less than .005 and P less than .05, respectively). The effect of the type of oil and the interactions tested (AB, AC, BC, ABC) were not statistically significant.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3047521 TI - Effect of improved glycemic control on the response of plasma triglycerides to ingestion of a saturated fat load in normotriglyceridemic and hypertriglyceridemic diabetic subjects. AB - This study tests the hypothesis that improved glycemic control decreases the postprandial plasma triglyceride (TG) response to ingestion of a saturated fat load. Fifteen normotriglyceridemic subjects with insulin-dependent diabetes mellitus (IDDM, group I) and six hypertriglyceridemic subjects with non-insulin dependent diabetes mellitus (NIDDM, group II) were studied. Each subject was studied before and after 12 days of continuous subcutaneous insulin infusion (CSII). Each subject ingested identical meals on both study days. Plasma glucose was determined in all patients before and two hours after each meal and at 3 AM, and a mean value was calculated for each patient. CSII reduced mean plasma glucose from 252 to 140 mg/dL in group I, and from 209 to 120 mg/dL in group II (P less than .001 in both groups, paired t test). Plasma TG levels were measured before and 1.5, 3, 4.5, 6, and 7.5 hours after a breakfast which contained 50 g of mostly saturated fat. A repeated-measures ANOVA was performed to assess the effects of glycemic control (factor A) and TG response (factor B) to fat ingestion. In both groups plasma TG levels increased significantly after fat ingestion (P less than .001), and were significantly reduced during improved glycemic control (P less than .001). The reduction was observed in 14 of 15 patients in group I and in all patients in group II. In group I the lowering of the postprandial plasma TG levels after CSII was secondary to a decrease in the fasting plasma TG levels, as shown by the unchanged mean percent TG elevation over the baseline.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3047523 TI - Hormonal effects of norepinephrine on acute glucose disposal in humans: a minimal model analysis. AB - It is not known whether circulating norepinephrine (NE) has a direct hormonal influence on glucose disposal. This study examines whether moderate elevation of NE alters the disposal of an acute intravenous (IV) glucose load, as analysed by the minimal model of Bergman. Eight healthy normal subjects were infused with either 25 ng/kg/min NE (plasma NE 1,284 +/- 259 pg/mL) or normal saline (plasma NE 314 +/- 86 pg/mL), 30 minutes prior to and during an IV glucose tolerance test (GTT). There was a small but significant rise (P less than .05) in basal blood glucose levels during the initial 30-minute NE infusion which was accompanied by a 40% increase (0.39 +/- .02 to 0.59 +/- .07 nmol/L, P less than .01) in nonesterified fatty acid levels (NEFA). Insulin, C-peptide, and glucagon levels did not change. NE impaired the rate of acute glucose disposal (Kg 1.74 +/- 0.24 v 2.10 +/- 0.23 (min-1, P less than .05). Minimal model analysis revealed a corresponding 35% decrease in insulin sensitivity (SI 4.85 +/- 1.51 v 7.28 +/- 1.16 min-1 microU-1 mL-1 x 10(4), P less than .05) but no significant differences between glucose-mediated glucose disposal or pancreatic B-cell responsiveness. The glucose disposition index (si* phi2), a direct measure of an individual's overall insulin- mediated glucose disposal, was reduced by 70% in the NE-infussed subjects (si* phi2 69 +/-22 v 223 +/- 76 mg-1 ml-1 min-3 x 10(2), p< .05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3047522 TI - First-phase insulin response to glucose in nonobese or obese subjects with glucose intolerance: analysis by C-peptide secretion rate. AB - This study was proposed to clarify the impairment of first-phase insulin response to glucose in subjects with glucose intolerance by analysis of C-peptide secretion rate after glucose or glucagon injection. The rate was calculated from kinetic analysis of peripheral C-peptide behavior. The rate reached the peak two minutes after glucose injection and then rapidly declined (first-phase secretion) in control subjects. In nonobese subjects with impaired glucose tolerance (IGT) or non-insulin-dependent diabetes mellitus (NIDDM), the rate promptly increased in response to glucose and was followed by a second phase increase. The time course of the rate in the subjects was slightly different from that in control subjects. There was a progressively greater deficit in the first-phase increase with increasing severity of glucose intolerance. The time course of the rate in the obese subjects with NIDDM was different from that in control subjects. The first-phase increase was reduced in the obese subjects with NIDDM. The glucose disappearance rate was correlated with the first-phase increase. Since the time course of the rate after glucagon injection in all subjects did correspond well with that in the control subjects, variation of metabolic clearance rate of endogenous C-peptide among the subjects may be negligible for this study. This study provides the precise time course of first- and second-phase insulin response to glucose injection in nonobese and obese subjects with IGT or NIDDM as well as convincing evidence of the progressive reduction of first-phase insulin response with increasing severity of glucose intolerance. First-phase insulin response to glucose might be slightly delayed in some obese subjects with NIDDM. PMID- 3047524 TI - Pancreatic endocrine function in cirrhotic rats. AB - Pancreatic endocrine function in liver cirrhosis was examined in rats both in vivo and in vitro. Experimental liver cirrhosis was induced by subcutaneous injections of 50% carbon tetrachloride in a dose of 2 mL/kg body weight twice a week for 16 weeks. Control rats received a similar dose of olive oil during the same period. In cirrhotic rats, immunoreactive insulin contents in the pancreas were significantly lower, whereas immunoreactive glucagon contents were about threefold higher than those of control rats. In the first part of this study, insulin and glucagon concentrations in both jugular and portal venous blood at basal conditions and after oral glucose loading were simultaneously determined in vivo. Peripheral insulin levels, both before and after glucose loading, were higher, whereas portal insulin concentrations were lower in cirrhotic rats than in the control rats. In contrast, glucagon levels in both the peripheral and portal veins were significantly higher in cirrhotic rats than in control rats. In the second part, isolated perfused pancreata were prepared from cirrhotic and control rats to further characterize the endocrine function of cirrhotic rat pancreas. Insulin secretion in response to 16.7 mmol/L glucose and 100 pmol/L cholecystokinin-octapeptide both were 40% lower in cirrhotic rats than in controls. In contrast, there was no significant difference in arginine-stimulated insulin release between the two groups. However, glucagon secretion in response to 20 mmol/L arginine was 40% higher in cirrhotic rats. If sensitivity is defined as the hormone release proportional to the pancreatic contents, then A and B cells in the cirrhotic rats had normal sensitivity to both glucose and cholecystokinin-octapeptide.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3047525 TI - Acute effects of S-carboxymethylated human growth hormone on insulin resistance in the obese (ob/ob) mouse. AB - Chronic treatment of ob/ob mice with growth hormone (GH) increases plasma insulin and blood glucose concentrations, and enhances insulin resistance in peripheral tissues. The purpose of the present study was: (1) to determine the length of time required for the development of increased circulating insulin concentration and adipose tissue insulin resistance in response to GH in the ob/ob mouse, (2) to examine the relationship between the rise in insulin concentration and the development of insulin resistance, and (3) to test whether the hormone derivative could enhance insulin resistance in isolated adipose tissue when added in vitro. Female ob/ob mice were injected intraperitoneally (ip) with either saline or 200 micrograms of S-carboxymethylated human GH (RCM-hGH), a diabetogenic GH derivative, which lacks significant insulinlike or growth-promoting activities. A threefold increase in plasma insulin concentration was observed three and six hours after RCM-hGH injection, but increased hyperglycemia was evident only after six hours. The in vitro stimulatory effect of insulin on [14C]glucose oxidation by parametrial adipose tissue was unchanged at three hours but suppressed six hours following administration of RCM-hGH. When the plasma insulin level of ob/ob mice was increased threefold by the administration of neutral protamine Hagedorn (NPH) insulin, the in vitro stimulatory effect of insulin on [14C]glucose oxidation by adipose tissue isolated from these animals was not altered, suggesting that insulin-induced receptor or postreceptor changes do not account for the increased insulin resistance produced by RCM-hGH.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3047526 TI - Comparison of two equine oestrogen-dydrogesterone regimens in the climacteric. AB - Menopausal women receiving 0.625 mg/day conjugated oestrogen continuously for 6 mth were also given either 10 mg/day dydrogesterone continuously or 20 mg/day for the first 12 days of each calendar month in order to evaluate both the efficacy and the tolerance of each regimen. The following parameters were assessed: subjective symptoms, bleeding patterns, lipid metabolism, glycaemia, weight and blood pressure. Of the 81 patients who entered the study, 60 opted for the continuous treatment and 21 for the cyclic treatment. Seven (7) patients dropped out, 6 for administrative reasons and 1 because of nausea. The two groups were comparable with regard to all parameters, with the exception of the duration of pretreatment amenorrhoea, which was longer in the continuous-treatment group (40.6 vs. 19.1 mth), and the initial lipid profile levels, which were higher in the same group. The subjective symptoms were influenced rapidly and positively by both treatments. The other parameters remained unchanged, except for a clinically insignificant rise in triglycerides in both groups. Both treatments were quite satisfactorily accepted by the patients, in spite of a 70% rate of withdrawal bleeding in the cyclic group and a 40% rate of spotting in the continuous treatment group. Sixty-two (62) patients remained free of side effects, while mastodynia was reported in 9 cases. PMID- 3047527 TI - Menopause and menorrhagia: a historical exploration. AB - Like a parallel article [1] this one also attempts to answer the question why complaints of excessive premenopausal bleeding have now virtually disappeared. Unlike the first, a review, this is a discussion paper. It explores widely, investigating the question from all aspects, even utilizing, if with caution, such unconventional sources as still unproven hypotheses or marginal theories. Presentational attitudes to, and handling of, climacteric complaints appear partially to obscure some symptoms associated with excessive menses. More importantly, however, the examination of the historical records in the light of present knowledge greatly increases our understanding of past socio-clinical events. It is then that attention is concentrated on a significant omission in these data: the fall in numbers of births per woman, first among the French and European aristocracy, then the population of France and finally that of the rest of Europe. This parallels the spread of the menopausal syndrome and complaints of excessive menstrual loss. It is suggested that limitation of the family, by design or secondarily to lifestyle, has created a unique bio-endocrinology in the West. In the past this produced a number of complications, including climacteric menorrhagia. Never redressed by preventative measures--for these had repeatedly been rejected as militating against the lifestyle chosen by Western women--they were only solved with the advent of abdominal hysterectomy. Steroidal oral contraception and hormonal treatment have, however, made this operation often unnecessary. PMID- 3047528 TI - Highlights from the history of caesarean section. PMID- 3047529 TI - Constituents of a "balanced" diet. PMID- 3047531 TI - The history of the use of vacuum extraction. PMID- 3047530 TI - To promote breeding from Tutankhamen to today. PMID- 3047532 TI - [Preservation of inoculation cultures of microorganisms at low temperatures]. PMID- 3047533 TI - [A rapid method for determining the viability of Escherichia coli cells subjected to the action of low temperatures]. AB - When Escherichia coli cells are frozen at a low temperature, various damages appear in them, in particular, in the membranous apparatus. Only stable disorders in the barrier properties of the cytoplasmic membranes are of a critical importance for the cell viability. These disorders should be taken into account when express methods are developed for assaying the viability of bacterial cells. Critical structures (properties) are to be revealed by their analysis after varying the intensity (dose) of the main damaging factors. Provided that the critical feature of each cell has two states (damaged and intact) after the action of extreme factors, its quantitative change correlates in a linear mode with the number of viable cells in the population. PMID- 3047535 TI - Presymptomatic testing for myotonic dystrophy by means of the linked DNA marker APOC2. AB - A family in which the gene for myotonic dystrophy is segregating was tested with a DNA probe to the apolipoprotein-C2 (APOC2) gene. The APOC2 gene is linked closely to the myotonic dystrophy (DM) gene and can be used as a marker to follow the transmission of the DM gene within families. Results from DNA-marker studies were combined with information from clinical, ophthalmological and electromyographic examinations, with age-dependent penetrance and a recombinant frequency of 4% between the genes for myotonic dystrophy and apolipoprotein C2 being taken into account. The chance that an individual had inherited the gene for myotonic dystrophy was determined by means of the computer program, LINKAGE. This analysis altered dramatically the assessed risk that certain individuals in the family were carrying the DM gene. For four individuals in the family, the previous risks of 50% for each member were reduced to 0.9%, 0.9% and 0.2%, respectively, in three cases and increased to 91% in the fourth case. The importance of a complete clinical evaluation of potential gene carriers who donate blood for a DNA-linkage study is stressed. PMID- 3047536 TI - Relapse of vivax malaria after falciparum malaria. PMID- 3047534 TI - Screening for thyroid malignancy: the role of fine-needle biopsy. AB - Cytological examination of the aspirate of fine-needle biopsy is becoming more widely accepted as a screening test for malignancy in thyroid nodules. However, it tends to be used in addition to, rather than instead of, more traditional methods. In this five-year prospective evaluation we performed fine-needle biopsy in 618 euthyroid patients with nodular thyroid enlargement, 86% of whom also underwent radionuclide scans and 55% of whom underwent ultrasound scans. In 19% of patients fine-needle biopsy yielded insufficient material for diagnosis, and in 14% of patients normal follicular cells were found (which indicated that the clinical lesion was not sampled). To date, a histological diagnosis has been obtained in 258 (42%) patients, 44 of whom had malignancies. The results of the radionuclide and ultrasound scans did not alter the odds in favour of the detection of malignancy. The cytological diagnosis of malignancy was falsely positive in two patients and falsely-negative in four patients (three cases of which probably were sampling errors). If, in addition to overtly-malignant cells, atypical Hurthle cells and follicular neoplasms were considered to be potentially malignant, fine-needle biopsy alone had a sensitivity of 87% and a specificity of 72%. This good accuracy would be reduced by sampling failures, but a policy of operating on all patients with potentially-malignant cells, or on those in whom satisfactory aspirates could not be obtained, would yield high rates of the diagnosis of malignancy and would reduce the number of operations. Our data indicate that the most-appropriate method of screening thyroid nodules for malignancy is fine-needle biopsy without pertechnetate scanning or ultrasound examinations. PMID- 3047537 TI - Autoantibodies and circulating immune complexes in subjects infected with human immunodeficiency virus. AB - We wish to report the results of a retrospective study of patients with acquired immunodeficiency syndrome (AIDS) and AIDS-related complex (ARC) as well as a group of asymptomatic human immunodeficiency virus (HIV)-infected individuals, in whom we examined markers of systemic autoimmune disease including anti-cell antibodies, immunoglobulin (Ig) and complement (C3 and C4) profiles as well as circulating immune complexes (CIC). Antibodies to cytoplasmic (but not nuclear) antigens were significantly elevated in the AIDS and ARC groups when compared with a population of normal individuals (for AIDS, P = 0.0002 vs. control; for ARC, P = 0.0004 vs. control). The non-AIDS/ARC, HIV+ group did not demonstrate significance (P = 0.054). Each of the three study groups also demonstrated increased Ig (P less than 0.05 for each immunoglobulin class). CIC, as determined by a Clq-binding enzyme immunoassay, were higher in all three study groups (P less than 0.05) when compared with controls. C3 and C4 were not significantly lower than control subjects (P greater than 0.05), but C4 did demonstrate a significant (P = 0.01) inverse correlation with CIC. These findings strengthen the hypothesized autoimmune aspect of AIDS and ARC, and extend this concept to include HIV-infected individuals without frank disease. PMID- 3047538 TI - Frequent detection of antibodies to hepatitis B virus x-protein in acute, chronic and resolved infections. AB - Recombinant MS2- or beta gal fusion proteins containing parts of hepatitis B virus (HBV) HBx-, HBc-, and HBs-amino acid sequences were expressed in Escherichia coli and were used to screen 96 and 60 serum samples of HBV infected and uninfected patients, respectively, for the corresponding antibodies by immunoblotting. Antibodies against HBx were detected in 20 out of 65 sera of patients with previous resolved HBV-infection, in 3 out of 7 patients with persistent infection, and in 9 out of 24 sera of patients with acute HBV infection. The specificity of the immune reaction was confirmed by competition experiments with MS2- and beta gal-HBx fusion proteins, and by the lack of HBx antibodies in the sera of uninfected patients. Hbs and HBc antibodies were detected less frequently by immunoblotting with recombinant fusion proteins than by a commercial immunoassay. Our results indicate that HBx antibodies are induced early and frequently during HBV infection suggesting that the HBx protein is an early antigenic protein expressed in vivo. PMID- 3047540 TI - Subacute myelopathy: an unusual paraneoplastic complication of Hodgkin's disease. AB - We report a case of Hodgkin disease presenting with a subacute myelopathy without evidence of metastatic involvement of the spinal cord. The systemic disease responded to conventional chemotherapy, but the myelopathy only improved after intrathecal dexamethasone was added to the treatment program, beta-2 microglobulin levels in the cerebrospinal fluid were elevated at presentation. Following the use of intrathecal corticosteroids there was a decrease of CSF beta 2-microglobulin levels. The possible significance of these findings is discussed. PMID- 3047541 TI - Infection of the central nervous system in organ transplant recipients. AB - Infections of the central nervous system are an important cause of morbidity and mortality in recipients of organ transplants. Three organisms--Listeria monocytogenes, Aspergillus fumigatus, and Cryptococcus neoformans--account for the great majority of such infections. The incidence of these infections is directly related to the patient's net state of immunosuppression and to the epidemiologic exposures he encounters. There is an expected timetable of when particular infections are likely to occur post-transplant. The most important factor in effecting patient survival is the rapidity with which diagnosis is made. Because of the nature of the infecting organisms and the impaired inflammatory response, the clinical presentation of CNS infection is often insidious, making early diagnosis difficult. However, an aggressive approach based upon lumbar puncture and CT scanning for patients with headache and fever or altered states of consciousness can be very rewarding. In addition, the use of such ancillary clues as the results of skin and lung biopsies can be an important clue to disseminated infection, particularly involving the CNS. The recent deployment of the MRI scan in immunosuppressed patients with unexplained CNS symptoms holds particular promise for early diagnosis. PMID- 3047539 TI - Modulating action of Escherichia coli heat-stable enterotoxin (STa) on the humoral immune response. AB - The effect of Escherichia coli enterotoxin STa on the primary and secondary immune response in F1 (CBA x C57 B1/10) mice immunized against sheep red blood cells (SRBC) was investigated. Modulating action on the IgM and IgG response was found to be dependent on the dose-time administration of the toxin. Immunosuppression of the primary response on the 4th day after immunization was observed when the toxin was injected 15 min before the SRBC, followed by immunostimulation on the 6th day after antigen (Ag) injection. Moreover, toxin administration 48 h before SRBC caused immunosuppression of the primary immune response on the 4th and 6th days. On the other hand, the IgM and IgG secondary immune response, determined 6 days after boosting, was greatly enhanced by toxin administration 15 min before priming (day 0) or boosting (day 26) and 48 h before priming. The same response was suppressed by toxin administration 48 h before booster antigen injection. PMID- 3047542 TI - Seizures and antiepileptic drug use in transplant patients. AB - Seizures may occur as an isolated manifestation of an acute encephalopathy or, less frequently, recur as a manifestation of epilepsy in transplant patients. Determining which of these is the case will lead to appropriate treatment. The selection of the antiepileptic drug (AED) will depend on the patient's type of seizure, general medical condition, and transplant type and also depend on the consideration of potential adverse effects of individual AEDs. PMID- 3047544 TI - Neurologic complications of liver transplantation. AB - The clinical and neuropathologic findings of 55 adults and 30 children who received liver transplants were reviewed. Encephalopathy was the most common clinical neurologic syndrome and was usually caused by metabolic or anoxic causes. (Alzheimer type astrocytes were present in 73 per cent of patients, and evidence of diffuse hypoxic damage was present in 40 per cent of children and 25 per cent of adults.) Cerebrovascular lesions were a common finding with infarcts or hemorrhages present in 30 per cent of patients. CNS infections were documented in 34 per cent of patients. Seizures were present in a third of patients. Central pontine myelinolysis was present in 12 per cent of patients and was more common in adults than in children. Antemortem diagnosis of neurologic complications was more often based on clinical presentation rather than specific radiologic or laboratory tests. PMID- 3047543 TI - Neurologic problems in renal transplant recipients. AB - Renal transplant recipients are at an increased risk of developing certain neurologic problems. These problems differ from those encountered in recipients of other organs. Development of atherosclerosis is accelerated in renal transplant patients and results in an increased incidence of thromboembolic events. Immunosuppressive therapy predisposes to infection with opportunistic organisms, including reactivation of latent viruses and also is associated with an increased incidence of de novo neoplasia. The transplantation procedure may be complicated by a neuropathy and occasionally by distal spinal cord infarction. Some immunosuppressive agents have a direct adverse effect on the nervous system, particularly when toxic levels accumulate in the body. Uremia prior to, and if present after transplantation, has a toxic effect on the nervous system as well. The reasons for these problems are discussed. Awareness of these special problems in renal transplant patients will facilitate their prevention and diagnosis. A recommended diagnostic approach has been outlined. After the etiology has been established, treatment can be tailored to the individual patient. PMID- 3047546 TI - Neurologic complications of pancreas transplants. AB - A review of 15 cases of pancreas transplantation at the Presbyterian University Hospital in Pittsburgh showed that all of the neurologic complications occurred outside of the pancreas transplantation surgery itself. Major CNS complications included hypoxic encephalopathy (20 per cent), cerebral and spinal-cord infarction (7 per cent), and seizures (13 per cent). These appeared to be closely associated with cardiovascular collapse or cardiac arrest that often occurred following septic, hemorrhagic, or additional surgical-anesthetic stresses, removed in time from the transplantation. When patients who died of sudden cardiorespiratory arrest were included, the overall frequency of global cerebral ischemia was 33 per cent. The occurrence of herpes zoster neuritis (13 per cent) was contrasted with the lack of CNS infections. The possible associations of visual hallucinations with cyclosporine therapy (7 per cent), CSF pleocytosis with OKT3 therapy (7 per cent), and compressive neuropathy with operative anesthetic monitoring (7 per cent) were discussed in relation to previous reports in the literature. Randomized controlled clinical studies were suggested to distinguish more clearly the complications due to pancreas transplantation from those due to the natural history of the underlying diabetes and to distinguish the beneficial and adverse effects of pancreas transplants from those of coexisting renal transplants. PMID- 3047545 TI - Neurologic complications of cardiac transplantation. AB - The neurologic evaluation of an individual cardiac transplant recipient often does not lead to a succinct bedside diagnosis. There are few consistent clinical observations. The onset of seizures in the early postoperative period is associated with embolic cerebral infarction. Seizures occur most commonly, however, as a neurotoxic manifestation of cyclosporine. The onset of an acute delirium or psychosis in the first week after cardiac transplantation usually has multiple causative factors and is reversible. A postoperative brachial plexopathy or mononeuropathy can be identified with a neurologic examination, confirmed by appropriate electrophysiologic testing and is usually reversible. The onset of periorbital inflammation, ophthalmoplegia, and nasal turbinate or sinus invasion and necrosis is consistent with phycomycosis. Most patients, however, present with nonspecific findings of impaired mentation with or without focal neurologic signs. These patients require a fairly systematic search for potentially treatable neurologic complications (see Table 3). In a medically stable patient an aggressive diagnostic approach, at times including stereotaxic brain aspirate or biopsy, is indicated. In the severely ill patient with multiple organ failure, empirical therapy for the most probable treatable disorder is justified. PMID- 3047547 TI - Neurologic complications of bone marrow transplantation. AB - Neurologic complications are extremely common after bone marrow transplantation and occur in well over half of all patients. Approximately 6 per cent of BMT recipients die as a direct result of neurologic problems. Metabolic encephalopathy, the most common clinical syndrome, is usually due to multiple organ failure. The second most common complication is CNS infection with fungi and viruses. Cerebrovascular disorders are the third most common neurologic problem, and most are related to underlying endocarditis (either infectious or nonbacterial thrombotic endocarditis). Less common neurologic complications include side effects of drugs, recurrence of malignancy, and treatment-induced leukoencephalopathy. Neurologic involvement due to GVHD appears to be limited to rare neuromuscular syndromes. No evidence of CNS involvement from GVHD has been detected. PMID- 3047548 TI - Neurologic complications of graft-versus-host disease. AB - BMT has become an important therapy for many hematologic disorders. Following BMT, the recipient may develop GVHD when it appears that immunocompetent donor lymphocytes react to host antigens. Acute and chronic GVHD represent two distinct syndromes. Acute GVHD has not been associated with primary neurologic involvement. Polymyositis has been reported in 12 patients with chronic GVHD, with the most common underlying illness being aplastic anemia. The clinical, serologic, and muscle biopsy features of the myositis in GVHD have been similar to those observed in idiopathic polymyositis. Weakness was moderate to severe and responded to prednisone, sometimes with the addition of azathioprine. Prognosis depended upon the underlying disease and not on the severity of the myositis. MG occurs rarely in chronic GVHD. Most patients with MG and GVHD have had aplastic anemia; those with aplastic anemia are more likely to have anti-AchR prior to BMT. The clinical manifestations of GVHD MG have not differed from classic autoimmune MG; each patient had elevated antiacetylcholine receptor antibodies titers. All patients have responded well to cholinesterase inhibitors but have received other immunosuppressants. These observations suggest that aplastic anemia is an important host factor in the development of the autoimmune disorders seen with chronic GVHD, certainly of myositis and MG. Herpes zoster peripheral nerve infections have occurred in patients with chronic GVHD. One patient had mononeuritis multiplex. In both acute and chronic GVHD, CNS impairment is usually caused by metabolic encephalopathy or infection. Primary CNS involvement has not been recognized. PMID- 3047549 TI - Brain transplants. A new approach to the therapy of neurodegenerative disease. AB - There is now a wealth of experimental evidence to suggest that transplantation to the brain may ameliorate a variety of neurologic and endocrine disorders. Many unanswered questions remain. Chief among these questions are the duration of any salutary effects and the potential long-term risks to the host CNS. Answers to these questions will only come with carefully controlled long-term clinical studies. Given the high incidence and devastating nature of many of these diseases, such studies will have enormous scientific and social impact. Regardless of the outcome, there is the potential for a greater understanding of the pathologic mechanisms underlying neurodegenerative diseases and, thus, the possibility that definitive therapies will be found as a result. PMID- 3047552 TI - Inhibition of gene expression of T7-related phages by prophage P1. AB - The gene expression of nine phages of the T7 group was compared after infection of Escherichia coli B(P1). With the exception of phage 13a which grew normally, all of them infected E. coli B(P1) abortively. Differences were found in the efficiency of host killing which ranged from 100% for phage 13a to 37% for phage A1122. Infection by T7 prevented colony formation by about 70% of the cells but they showed filamentous growth until about 2 h after infection. It was shown by SDS-polyacrylamide gel electrophoresis and autoradiography of [35S]methionine labelled phage-coded proteins that all phages except for 13a showed measurable expression only of the early genes. No correlation was observed between killing capacity and the pattern of gene expression, and the ability to hydrolyse S adenosyl-methionine (SAM, a cofactor for the P1 restriction endonuclease) by means of a phage-coded SAMase. Mixed infection of E. coli B(P1) with 13a and T7 yielded mixed progeny indistinguishable from that observed after mixed infection of the normal host E. coli B. Genetic crosses with amber mutants of 13a and T7 showed that the 13a marker opo+ (overcomes P one), required for growth on B(P1), is located in the early region, to the left of gene 1 (RNA polymerase gene). PMID- 3047550 TI - Genetic mapping of the Saccharomyces cerevisiae DNA polymerase I gene and characterization of a pol1 temperature-sensitive mutant altered in DNA primase polymerase complex stability. AB - The cloned DNA polymerase I gene has been used to map the POL1 locus on the left arm of chromosome XIV, between MET4 and TOP2. Temperature-sensitive mutants in POL1 have been obtained by in vitro mutagenesis of the cloned gene and in vivo replacement of the wild-type allele with the mutated copy. Physiological and biochemical characterization of one temperature-sensitive mutant (pol1-1) shows that cells shifted to the non-permissive temperature can complete one round of cell division and DNA replication before they arrest. Analysis of DNA polymerase I in crude extracts and in partially purified preparations indicates that the pol1-1 mutation results in a conformational change and affects the stability of the DNA primase-polymerase complex. PMID- 3047551 TI - A consensus transcription termination sequence in the promoter region is necessary for efficient gene expression of the TRP1 gene of Saccharomyces cerevisiae. AB - The TRP1 gene of Saccharomyces cerevisiae is the only TRP gene which is not derepressible by the general control regulatory system. In the TRP1 promoter transcription starts at five initiation sites, organized in two clusters. The two transcripts of the first, more upstream cluster include a long leader sequence of approximately 200 bp. A transcriptional terminator element located in the 5' region of the TRP1 gene is essential for accurate gene expression. In partial TRP1 promoters lacking the terminator, like the original EcoRI TRP1 fragment used in numerous vectors, plasmid-encoded transcription is initiated predominantly in adjacent vector regions, resulting mainly in large, poorly translated transcripts. This poor translation is not due to mRNA instability. The effect can be suppressed by introducing artificial transcription barriers between vector sequences and the truncated EcoRI TRP1 fragment. PMID- 3047553 TI - Characterization of agonist radioligand interactions with porcine atrial A1 adenosine receptors. AB - The agonist radioligand (-)-N6-[125I]-p-hydroxyphenylisopropyl-adenosine ([ 125I]HPIA) was used to characterize adenosine recognition sites in porcine atrial membranes. [125I]HPIA showed saturable binding to an apparently homogeneous population of sites with a maximum binding capacity of 35 +/- 3 fmol/mg of protein and an equilibrium dissociation constant of 2.5 +/- 0.4 nM. Kinetic experiments were performed to address the molecular mechanism of [125I]HPIA binding in porcine atrial membranes. [125I]HPIA apparently interacts with the cardiac adenosine receptor in a simple bimolecular reaction. A kinetically derived [125I] HPIA dissociation constant (2.4 nM) was in good agreement with that parameter measured at equilibrium. Guanyl nucleotides negatively modulated [125I]HPIA binding by increasing its rate of dissociation. This finding is consonant with the formation of a ternary complex in porcine atrial membranes, consisting of ligand, receptor, and guanyl nucleotide-binding protein. Prototypic adenosine receptor agonists and antagonists inhibited specific binding in a manner consistent with the labeling of an A1 adenosine receptor. The results of these experiments suggest that the adenosine receptor present in porcine atrial membranes, as labeled by [125I]HPIA, is of the A1 subtype. PMID- 3047555 TI - Characterization of proliferin-related protein. AB - Proliferin-related protein (mPRP) is a member of the PRL/GH family in the mouse. We have generated an antiserum against mPRP expressed as a bacterial fusion protein; this antiserum detects mPRP in the conditioned media of placental tissue cultures as a heterogeneous population of glycoproteins. We have also expressed mPRP in mammalian tissue culture cells and purified the secreted protein. N terminal sequence analysis of the purified protein reveals that it is secreted as a 214 amino acid protein after removal of a 30 amino acid signal polypeptide. An antiserum raised against the purified protein detects high levels of mPRP in maternal serum during gestation. The site of synthesis of this protein has been localized by in situ hybridization to the basal zone of the day-10 mouse placenta, which is distinct from the site of synthesis of other placental proteins in this family. PMID- 3047556 TI - Cytopathology of pulmonary disease. PMID- 3047554 TI - Expression of transforming growth factor alpha and its messenger ribonucleic acid in human breast cancer: its regulation by estrogen and its possible functional significance. AB - We have studied the estrogenic regulation and the potential autocrine role of transforming growth factor alpha (TGF alpha) in the human breast cancer cell line MCF-7. A biologically active apparent mol wt 30 k TGF alpha was identified by gel filtration chromatography in medium conditioned by MCF-7 breast cancer cells. We previously reported induction of TGF alpha levels in medium by 17 beta-estradiol. We now report correlated increases in TGF alpha mRNA, by Northern and slot blot analysis, after estrogen treatment of MCF-7 cells in vitro. In vivo experiments confirmed these data: estrogen withdrawal from MCF-7 tumor-bearing nude mice resulted in a decline in tumor size and TGF alpha mRNA levels. To explore the functional significance of TGF alpha in MCF-7 cells, anti-TGF alpha antibody was added to MCF-7 soft agar cloning assays. Inhibition of MCF-7 growth resulted, supporting an autocrine role for TGF alpha. Further experiments using an anti-EGF receptor antibody expanded this data, demonstrating inhibition of estrogen stimulated monolayer MCF-7 cell growth. Examining the generality of TGF alpha expression, 4.8 kilobase TGF alpha mRNAs were seen in three other human breast cancer cell lines, MDA-MB-231, ZR 75B, and T47D. Expression of TGF alpha mRNA was detected in 70% of estrogen receptor positive and negative primary human breast tumors from 40 patients when examined by slot blot and Northern analysis. Thus, we have demonstrated broad expression of TGF alpha in human breast cancer, its hormonal regulation in an estrogen-responsive cell line, and its possible functional significance in MCF-7 cell growth. PMID- 3047557 TI - [Biological principles of psychological development in infancy]. PMID- 3047558 TI - [Developmental directions in organ transplantation and review]. PMID- 3047559 TI - [Bone marrow transplantation in children and adolescents]. PMID- 3047560 TI - [Organ procurement from children for children]. PMID- 3047561 TI - [Dialysis procedures and indications for kidney transplantation in childhood]. PMID- 3047562 TI - [Kidney transplantation in childhood]. PMID- 3047563 TI - [Indications and results of liver transplantation in childhood]. PMID- 3047564 TI - [Surgical aspects and postoperative complications following liver transplantation in childhood]. PMID- 3047565 TI - [Orthotopic heart transplantation in childhood and adolescence]. PMID- 3047566 TI - [Personal experiences with pancreas transplantation]. PMID- 3047567 TI - Current status of cytogenetic procedures to detect and quantify previous exposures to radiation. AB - The estimation of the magnitude of a dose of ionizing radiation to which an individual has been exposed (or of the plausibility of an alleged exposure) from chromosomal aberration frequencies determined in peripheral blood lymphocyte cultures is a well-established methodology, having first been employed over 25 years ago. The cytogenetics working group has reviewed the accumulated data and the possible applicability of the technique to the determination of radiation doses to which American veterans might have been exposed as participants in nuclear weapons tests in the continental U.S.A. or the Pacific Atolls during the late 1940s and 1950s or as members of the Occupation Forces entering Hiroshima or Nagasaki shortly after the nuclear detonations there. The working group believes that with prompt peripheral blood sampling, external doses to individuals of the order of about 10 rad (or less if the exposure was to high-LET radiation) can accurately be detected and measured. It also believes that exposures of populations to doses of the order of maximum permissible occupational exposures can also be detected (but only in populations; not in an individual). Large exposures of populations can also be detected even several decades after their exposure, but only in the case of populations, and of large doses (of the order of 100 to several hundred rad). The working group does not believe that cytogenetic measurements can detect internal doses from fallout radionuclides in individuals unless these are very large. The working group has approached the problem of detection of small doses (less than or equal to 10 or so rad) sampled decades after the exposure of individuals by using a Bayesian statistical approach. Only a preliminary evaluation of this approach was possible, but it is clear that it could provide a formal statement of the likelihood that any given observation of a particular number of chromosomal aberrations in a sample of any particular number of lymphocytes actually indicates an exposure to any given dose of radiation. It is also clear that aberration frequencies (and consequently doses) would have to be quite high before much confidence could be given to either exposure or dose estimation by this method, given the approximately 3 decades of elapsed time between the exposures and any future blood sampling.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3047568 TI - 2-Nitrofluorene and related compounds: prevalence and biological effects. PMID- 3047569 TI - Post-replication repair and recombination in uvrA umuC strains of Escherichia coli are enhanced by vanillin, an antimutagenic compound. AB - Effects of vanillin on UV killing of umuC mutant strains of E. coli were investigated in order to analyze the antimutagenic role of vanillin in mutagenesis. UV-irradiated uvrA umuC cells showed higher survival when plated on medium containing vanillin rather than medium without vanillin. This increased survival associated with exposure to vanillin was observed more clearly in uvrA umuC lexA(Ind-) and uvrA umuC recF strains. However, the effect was inhibited by additional recB recC mutations and completely blocked by an additional recA mutation. As far as tested the increased survival of UV-treated cells by vanillin was dependent on a capacity for genetic recombination. The effect of vanillin on recombination frequency between 2 plasmid DNA, pATH4 (Cmr Tcs) and pBMX7 (Apr Tcs), in a uvrA umuC background was investigated. A significantly higher frequency of plasmid recombination was observed when vanillin was present in the culture medium. These findings suggest that the antimutagenic effect of vanillin is the result of enhancement of a recA-dependent, error-free, pathway of post replication repair. PMID- 3047570 TI - N-nitroso-N-2-fluorenylacetamide: a new direct-acting mutagen and teratogen. AB - Reaction of N-2-fluorenylacetamide (2-FAA, CAS No. 53-96-3) with nitrous fume (N2O3) in glacial acetic acid at 0 degree C yields N-nitroso-N-2 fluorenylacetamide (N-NO-2-FAA), 3-nitro-N-2-fluorenylacetamide, N-nitroso-3 nitro-N-2-fluorenylacetamide and other compounds. N-NO-2-FAA is the major product (80%) and fairly stable at low temperature (-20 degrees C), but extremely labile at ambient temperature. The chemical structure of N-NO-2-FAA is characterized by spectrometric analysis of its naphthol coupling derivatives. This new compound is highly mutagenic to Salmonella typhimurium TA97, TA98, TA100 and TA1538 and requires no microsomal metabolic activation. The mutagenicity of N-NO-2-FAA in TA98 is higher than that of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG, CAS No. 70-25-7) and N-acetoxy-N-2-fluorenylacetamide (N-AcO-2-FAA). The teratogenic potential of N-NO-2-FAA was studied with white Leghorn chick embryos given a single dose of 1-100 micrograms/egg on day 6 of incubation. A high incidence of flaccid paralysis of the legs and a low incidence of feather, claw and bill malformations were found in the treated group; no such malformed embryos were found in the control group. The teratogenicity of N-NO-2-FAA was found to be weaker than that of MNNG, but comparable to that of N-methyl-N-nitrosourea (CAS No. 684-93-5). N-NO-2-FAA is a strong electrophile and reacts readily with histidine, lysine, cysteine, glutathione, tryptophan, adenosine, cytidine at neutral pH. In contrast to N-AcO-2-FAA, N-NO-2-FAA does not react significantly with guanosine and thymidine. It seems that N-NO-2-FAA is a strong direct-acting mutagen and probably a new prototype of synthetic carcinogen. PMID- 3047571 TI - Pure exogenous singlet oxygen: nonmutagenicity in bacteria. AB - Singlet oxygen (1 delta gO2) is the lowest energy-excited state of molecular oxygen, and more reactive than the triplet ground-state molecule. Although singlet oxygen has been implicated in a variety of biological effects, including reactions with DNA or some of its components, evidence for mutagenesis by singlet oxygen has remained unclear. We have previously described a system for bacterial exposure to pure exogenous singlet oxygen that eliminates ambiguity regarding the identity of the reactive species responsible for observed results. Despite the potent toxicity of pure singlet oxygen for several different strains of bacteria, we have found no evidence for mutagenicity of singlet oxygen in 26 Salmonella typhimurium histidine-auxotrophic strains killed to 35% survival. These strains included a variety of base-pair substitution or frameshift target sequences for reversion, including targets responsive to oxidative damage and targets rich in GC base pairs. Some strains combined histidine mutations with one or more mutations affecting DNA-repair capacity. 4 strains possessing the hisG46 mutation also were not mutated when exposed to dose ranges killing less than 28% and up to 99% of the bacteria. The relative frequency of small inphase deletions was assayed in hisG428 bacteria exposed to single oxygen and found to be the same as the spontaneous level. In addition to lack of induction of mutation in these strains, the 8-azaguanine forward mutation assay yielded no evidence of mutagenesis by singlet oxygen in strains killed to 15% survival. No induction of genetic changes by singlet oxygen was seen in an assay for duplication of approximately 1/3 of the bacterial chromosome. Tests for the ability of singlet oxygen to induce lambda prophage in E. coli K12 also proved negative. These studies collectively indicate that pure singlet oxygen generated outside the bacterial cell does not react significantly with the bacterial chromosome in ways leading to base-pair substitutions, frameshift mutations, small or large deletions, large duplications, or damage that interferes with DNA replication and induces the SOS system. PMID- 3047572 TI - Phenotypic and reversion analysis of a Salmonella typhimurium constructed to have an arginine codon at the hisG46 missense codon. AB - Of the 6 single-base mutations that would be predicted to change the missense mutation hisG46 away from a proline codon in the Salmonella/microsome mutagen selection assay for histidine-independent revertants, only 5 have been observed. We have used site-specific mutagenesis to make the unobserved mutant [CCC (proline)----CGC (arginine)] codon in the Salmonella genome. Experiments with this arginine mutant demonstrate that, like bacteria containing the hisG46 mutation, bacteria with the arginine missense mutation are histidine auxotrophs which are capable of reversion to histidine independence. However, unlike the ATP phosphoribosyltransferase coded by the hisG46 his G gene (with a proline), the arginine mutant enzyme is partially active. This is indicated by a histidine independent phenotype when the arginine hisG gene is present in multiple copies. PMID- 3047573 TI - 'Petite' mutagenesis in Saccharomyces cerevisiae by a series of 2,7-di-alkyl substituted derivatives of proflavine with differing DNA-binding properties. AB - The ability of proflavine (3,6-diaminoacridine) and its 2,7-dimethyl, 2,7 diethyl, 2,7-diisopropyl and 2,7-di-tert.-butyl derivatives to induce the 'petite' mutation in Saccharomyces cerevisiae has been studied in relation to the DNA-binding properties of the compounds. The nature of the binding has been investigated by nuclear magnetic resonance techniques, and the results support and clarify earlier suggestions that the first 3 members of the series intercalate into DNA while the diisopropyl and di-tert.-butyl compounds do not. Toxicity of the drugs was primarily associated with their mode of DNA binding, but lipophilicity had an important secondary effect. It seems likely that the toxic properties of the more lipophilic DNA-intercalating members of the series mask their potential for 'petite' mutagenesis. PMID- 3047574 TI - N-methyl-N'-nitro-N-nitrosoguanidine-induced mutation in a RecA strain of Escherichia coli. AB - 274 N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)-induced forward mutations in the lacI gene of an Escherichia coli RecA- strain were cloned and sequenced. Base substitutions accounted for 264 mutations and consisted of 261 G:C----A:T transitions (including one double mutant with two G:C----A:T transitions separated by 25 base pairs), two A:T----G:C transitions and one A:T----T:A transversion. Therefore, 263 of the 274 mutations (all the transitions) can be explained as a result of the direct mispairing of O6-methylguanine, and O4 methylthymine residues during DNA synthesis. The source of the transversion is not known. The remaining mutations, one 16-base pair deletion, two -1 frameshifts and 7 frameshifts at the lacI frameshift hotspot, are located in runs of identical bases or flanked by directly repeated DNA sequences and can therefore be explained by template slippage events during DNA synthesis. The observed distribution of mutations recovered is identical to that found in a RecA+ background indicating little involvement of RecA function in MNNG-induced mutation. Analysis of neighbouring base sequence revealed that the G:C----A:T transition was 6 times more likely to be recovered if the mutated guanine residue was preceded by a purine rather than a pyrimidine. A most striking aspect of this distribution concerns particular residues in the core domain of the lac repressor protein. Within this domain the great majority of mutations generate nonsense codons or alter Gly codons. PMID- 3047576 TI - [Important aspects of the epidemiology of aspergillosis]. PMID- 3047575 TI - Study of the causes of direct-acting mutagenicity in coffee and tea using the Ara test in Salmonella typhimurium. AB - The mutagenic activities of 6 of the chemicals identified in coffee solutions were assayed with the Salmonella Ara test, under experimental conditions optimized for coffee mutagenicity. Caffeine was the only non-mutagenic compound. Among the other 5 chemicals, hydrogen peroxide was the strongest mutagen and chlorogenic acid the weakest; methylglyoxal, glyoxal and caffeic acid exhibited intermediate mutagenicities. The minimal mutagenic doses of these components correlated negatively with their relative concentrations in coffee. It was concluded that chlorogenic acid, caffeic acid, glyoxal and methylglyoxal cannot contribute alone to the mutagenicity of coffee in the Ara test, since their minimal mutagenic concentrations were much higher than their respective levels in the coffee samples assayed. By contrast, 40-60% of the mutagenic activity in coffee and also in tea could be attributed to their H2O2 contents. Catalase abolished more than 95% of the mutagenic activity of coffee, as detected by the Ara test. A similar sensitivity to catalase has been reported by other authors in relation to the coffee mutagenicity identified by the Salmonella His test. Nevertheless, the results presented in this paper suggest that the Ara forward and the His reverse mutation tests are sensitive to the mutagenicity of different constituents in coffee solutions. We propose that the His test, sensitive at high coffee doses, mainly recognizes the mutagenicity of methylglyoxal, whilst the Ara test, sensitive at low coffee doses, mainly detects the mutagenic activity of hydrogen peroxide. The data reported also suggest that the direct-acting mutagenicity(ies) detected by the Ara test in tea solutions is (are) based on similar, if not identical, mechanisms. PMID- 3047578 TI - How did Medicare's prospective payment system affect hospitals? (a further note) PMID- 3047577 TI - New therapeutic approach in skin mycoses: a comparative trial once versus twice daily application of fenticonazole in comparison to miconazole. PMID- 3047579 TI - Prophylactic sclerotherapy of large esophageal varices. PMID- 3047580 TI - Reimbursement for computer-assisted literature searches for patient care. PMID- 3047582 TI - The molecular genetics of Philadelphia chromosome-positive leukemias. PMID- 3047581 TI - Dexamethasone therapy for bacterial meningitis. Results of two double-blind, placebo-controlled trials. AB - We enrolled 200 infants and older children with bacterial meningitis in two prospective double-blind, placebo-controlled trials to evaluate the efficacy of dexamethasone therapy in addition to either cefuroxime (Study 1) or ceftriaxone (Study 2). Altogether, 98 patients received placebo and 102 received dexamethasone (0.15 mg per kilogram of body weight every six hours for four days). At the beginning of therapy, the clinical and demographic characteristics of the patients in the treatment groups were comparable. The mean increase in the cerebrospinal fluid concentration of glucose and the decreases in lactate and protein levels after 24 hours of therapy were significantly greater in those who received dexamethasone than in those who received placebo (glucose, 2.0 vs. 0.4 mmol per liter [36.0 vs. 6.9 mg per deciliter], P less than 0.001; lactate, 4.0 vs. 2.1 mmol per liter [38.3 vs. 19.8 mg per deciliter], P less than 0.001; and protein, 0.64 vs. 0.25 g per liter [64.0 vs. 25.3 mg per deciliter], P less than 0.05). One patient in the placebo group in Study 1 died. As compared with those who received placebo, the patients who received dexamethasone became afebrile earlier (1.6 vs. 5.0 days; P less than 0.001) and were less likely to acquire moderate or more severe bilateral sensorineural hearing loss (15.5 vs. 3.3 percent; P less than 0.01). Twelve patients in the two placebo groups (14 percent) had severe or profound bilateral hearing loss requiring the use of a hearing aid, as compared with 1 (1 percent) in the two dexamethasone groups (P less than 0.001). We conclude that dexamethasone is beneficial in the treatment of infants and children with bacterial meningitis, particularly in preventing deafness. PMID- 3047583 TI - Similarity of endothelin to snake venom toxin. PMID- 3047584 TI - The functional logic of cortical connections. AB - Patterns of anatomical connections in the visual cortex form the structural basis for segregating features of the visual image into separate cortical areas and for communication between these areas at all levels to produce a coherent percept. Such multi-stage integration may be a common strategy throughout the cortex for producing complex behaviour. PMID- 3047585 TI - Evolution of a bacteria/plasmid association. AB - Associations between bacteria and their accessory elements (viruses, plasmids and transposons) range from antagonistic to mutualistic. A number of previous studies have demonstrated that plasmid carriage reduces bacterial fitness in the absence of selection for specific functions such as antibiotic resistance. Many studies have demonstrated increased fitness of evolving microbial populations in laboratory environments, but we are aware of only one study in which fitness gains were partitioned between a plasmid and its host. Here, we examine the evolution of an association between a plasmid and its bacterial host. Carriage of the non-conjugative plasmid pACYC184 initially reduced the fitness of Escherichia coli B in the absence of antibiotic. We then cultured plasmid-bearing bacteria for 500 generations in the presence of antibiotic. The fitness of each combination of host and plasmid, with and without the culture history, was determined by competing it against a baseline strain. The results indicate adaptation by the host genome, but no plasmid adaptation. We also competed the evolved host, transformed with the baseline plasmid, against its isogenic plasmid free counterpart. The plasmid now increased the fitness of its host. PMID- 3047586 TI - First scientific fraud conviction. PMID- 3047587 TI - Evolution of new tRNA-synthetase classes. PMID- 3047588 TI - Spatial clumping of sexually receptive females induces space sharing among male voles. AB - The spatial organization of individuals in populations (their spacing system) can be highly variable even among populations of the same species. As spacing systems have important consequences for ecological processes such as population regulation, competition and mating systems, there have been many attempts to explore factors that may cause this variation. For mammals, it has been argued that the spatial distribution of sexually receptive females is the most important factor determining the spacing system of males, whereas habitat characteristics are most important to females. This has been difficult to test experimentally as it requires manipulations of the spatial distribution of the opposite sex without changing other properties of the environment. Here, I present a novel experimental procedure that can achieve this and demonstrate that the spatial distribution of the opposite sex in a population of voles is indeed an important determinant of the spacing system of males, but not of females. However, the effects on males are different from those predicted by many theoretical studies. PMID- 3047589 TI - Insulin stimulation of glucose uptake can be mediated by diacylglycerol in adipocytes. AB - An early effect of insulin in adipocytes is to stimulate glucose uptake. The increased uptake appears to be due to mobilization of glucose transporters from an intracellular location to the plasma membrane and to enhanced intrinsic activity of the transporters. Little is known about the insulin-generated signals causing these changes. Phorbol esters have been shown to mimic the insulin effect, but phosphorylation of the transporter does not seem to be involved. A phospho-oligosaccharide was recently shown to mimic the effects of insulin on protein phosphorylation, suggesting that it could be a mediator for some intracellular metabolic effects of the hormone, but it did not affect glucose uptake. A diacyglycerol is produced in the plasma membrane in conjunction with the generation of the phospho-oligosaccharide. Here I show that added 1,2 diacylglycerols potently increase glucose transporter-mediated uptake of glucose in rat adipocytes, but without activation of protein kinase C. PMID- 3047590 TI - GAL4-VP16 is an unusually potent transcriptional activator. AB - Recent work has defined a class of transcriptional activators, members of which activate transcription in yeast, plant, insect and mammalian cells. These proteins contain two parts: one directs DNA binding and the other, called the activating region, presumably interacts with some component of the transcriptional machinery. Activating regions are typically acidic and require some poorly-understood aspect of structure, probably at least in part an alpha helix. Here we describe a new member of this class, formed by fusing a DNA binding fragment of the yeast activator GAL4 to a highly acidic portion of the herpes simplex virus protein VP16 (ref. 11; also called Vmw65). VP16 activates transcription of immediate early viral genes by using its amino-terminal sequences to attach to one or more host-encoded proteins that recognise DNA sequences in their promoters. We show that the hybrid protein (GAL4-VP16) activates transcription unusually efficiently in mammalian cells when bound close to, or at large distances from the gene. We suggest that the activating region of VP16 may be near-maximally potent and that it is not coincidental that such a strong activator is encoded by a virus. PMID- 3047591 TI - Effects of peptide HI on basal and stimulated insulin and glucagon secretion in the mouse. AB - Peptide HI (PHI) is a peptide with 27 amino acids that is structurally similar to VIP (vasoactive intestinal peptide). Since PHI, like VIP, has been demonstrated to occur in intrapancreatic neurons, and since VIP earlier was shown to stimulate insulin and glucagon secretion in the mouse, we investigated whether also PHI affects islet hormone secretion in this species. PHI was thereby injected intravenously at dose levels between 0.5 and 8.0 nmol/kg. It was found that PHI did not affect basal levels of insulin of glucagon. However, a slight hyperglycemia was observed after injection of PHI at dose levels above 4.0 nmol/kg. When injected together with glucose (2.8 nmol/kg), PHI (1.0 and 4.0 nmol/kg) potentiated the insulin response by approximately 35% (P less than 0.05) and 50% (P less than 0.01), respectively. In contrast, the insulin response to the cholinergic agonist carbachol (0.16 mumol/kg) or the beta 2-adrenoceptor agonist terbutaline (3.6 mumol/kg) was not affected by PHI. The glucagon response to carbachol was potentiated by PHI (1.0 and 4.0 nmol/kg) by approximately 40% (P less than 0.05) and 55% (P less than 0.01), respectively, whereas the terbutaline induced increase in plasma glucagon levels was not affected by PHI. In summary, PHI potentiates glucose-induced insulin secretion and carbachol-induced glucagon secretion in the mouse. Since similar effects earlier have been demonstrated for VIP, it is concluded that PHI in this species exerts VIP-like effects. PMID- 3047592 TI - Anomalies and variations of the middle cerebral artery: a microanatomical study. AB - The microvascular anatomy of the main trunk and divisions of the middle cerebral artery was studied in 104 unfixed brain hemispheres injected with polyester resin and dissected under the operating microscope. The following anomalies and variations of the middle cerebral artery were found: fenestration (1 case; 1%), located on the first 4 mm of the main trunk of the middle cerebral artery; duplication (1 case; 1%), with vessels arising from the internal carotid artery; accessory middle cerebral artery (2 cases; 2%), originating on the A1 segment of the anterior cerebral artery; single-trunk type of middle cerebral artery (4 cases; 4%), with no division of its main trunk; quadrifurcation (4 cases; 4%), in which the main trunk of the middle cerebral artery divided into four secondary trunks. The clinical implications of these anatomical findings are discussed, and photographs of representative specimens illustrate the anomalies. PMID- 3047593 TI - Malignant growth hormone-secreting pituitary adenoma with hematogenous dural metastasis: case report. AB - A rare case of growth hormone-secreting pituitary adenoma with hematogenous metastasis to the dura mater of the cerebral convexity is presented. Immunohistological staining was essential to the diagnosis. The histological findings demonstrated that the metastasis was blood-borne. Extensive removal of the tumor and postoperative chemotherapy resulted in partial remission. The mechanism of metastasis, the histological findings, the treatment, and the prognosis are discussed. PMID- 3047594 TI - Bilateral cholesterol granuloma of the skull base: case report and review of the literature. AB - A unique case of bilateral cholesterol granuloma of the skull base and its treatment is presented. Cholesteatoma, a pathological entity often confused with cholesterol granuloma, is differentiated from cholesterol granuloma. Cholesterol granuloma is not rare. This tumor seems to derive from an inflammatory process at the skull base that results in bony erosion surrounding a cyst wall of inflammatory tissue. Neurological abnormalities reflect the location of the tumor in relation to the brain stem. Radiographically, the cyst wall enhances with the administration of i.v. contrast agent, and the center of the lesion is isodense with brain on computed tomography, unlike cholesteatoma. Magnetic resonance imaging characteristics are currently being defined. At operation, cholesterol granuloma consists primarily of a viscous fluid within a capsule of inflammatory tissue. Treatment requires establishing a pathway for drainage of the granuloma. The advantages of transsphenoidal, transclival drainage of such lesions are outlined. PMID- 3047595 TI - Chondroblastoma of the temporal bone. AB - A chondroblastoma within the temporal bone in a 16-year-old boy is reported. The tumor recurred after intracapsular resection and curettage. After reoperation, radiotherapy was performed. The data from 137 cases of chondroblastoma are summarized. PMID- 3047596 TI - Commitment and contribution. PMID- 3047597 TI - The American centennial of brain tumor surgery. AB - Early in 1887 the first well-documented total removal of an unequivocal brain tumor in America was carried out in New York City by Robert F. Weir. The patient died during the immediate postoperative period. Later in the same year in Philadelphia, W.W. Keen also removed a large meningioma; his patient lived many years. In 1886, a Dr. Morse in California had probably performed a partial removal of a glioma from a patient who died shortly after the operation. To celebrate the centennial of Keen's triumph, it is worthwhile as well to contemplate the efforts of his contemporaries. PMID- 3047598 TI - Controversies in the management of cerebral vascular disease. PMID- 3047599 TI - [In search of lost time]. PMID- 3047600 TI - Single dose fosfomycin trometamol (Monuril) for the treatment of women with bacterial cystitis. PMID- 3047601 TI - Art & science of management. Does anything ever change? PMID- 3047602 TI - Advanced extrauterine pregnancy: description of 38 cases with literature survey. AB - An analysis is presented of 38 patients with advanced extrauterine pregnancy. First three typical cases are described that emphasize the marked differentiation of clinical symptoms which these patients present to the doctor. The first patient was referred for induction because of a suspected intrauterine death. The second patient presented an intraligamentous pregnancy with a living fetus. In the third case, the patient was admitted to hospital after 32 weeks of pregnancy because of a persistent oblique lie. At 34 weeks, a normal living fetus was born. In all three cases, ultrasound examination was able to visualize the separate uterus. A literature survey is given with special attention to the specific "clinic" and the problems concerning diagnosis and treatment. It is obvious that sonography is the most important diagnostic technique at present. The decision to remove the placenta by means of a laparotomy is brought up for discussion. PMID- 3047603 TI - Borderline epithelial tumors of the ovary: ovarian intraepithelial neoplasia. AB - Although the concept of low malignant potential and/or borderline malignancy of some epithelial ovarian tumors was endorsed by the World Health Organization in 1973, uncertainty exists regarding the biologic behavior aspects of these lesions and this may account for the discrepancy in the 5-year survival figures reported for patients afflicted with these malignancies (76-95 per cent). We have reviewed the clinicopathologic aspects of 26 cases of borderline epithelial ovarian tumors and searched the literature. Based on our analysis, we have concluded that: 1) rupture of the cyst at surgery did not affect the patient's outcome but positive peritoneal fluid cytology did. 2) The term borderline should be replaced by ovarian intraepithelial neoplasia or preinvasive carcinoma and should solely be used in patients with stage I disease. 3) There is no justification for adjuvant therapy in adequately staged and surgically treated stage Ia and Ib disease. 4) Patients with stage II or more disease and those with positive peritoneal fluid cytology should be treated as aggressively as all other invasive, well differentiated, epithelial ovarian tumors. 5) Our observation in cases of epithelial ovarian tumor cells grown on an extracellular matrix tends to indicate that parameters other than morphology may aid in assessing the invasive potential of these malignancies. PMID- 3047604 TI - Obstetrical concerns of epidermolysis bullosa. AB - Epidermolysis bullosa is a group of autosomally inherited diseases involving skin breakdown of varying degrees and severity. Prenatal diagnosis and evaluation of these diseases is reviewed although pregnancy is not generally adversely affected by these diseases. Labor and delivery practices must be altered for the management of these patients. The obstetrical team must pay careful attention to adhesive dressings and anesthetic devices. Minor trauma may lead to severe lesions. PMID- 3047605 TI - Berger's renal disease in pregnancy: a case report and review of the literature. PMID- 3047606 TI - Maternal toxemia and neonatal germinal matrix hemorrhage in intubated infants less than 1751 g. AB - Two hundred seventy-two intubated infants who weighed less than 1751 g were enrolled in a clinical trial of phenobarbital prophylaxis of postnatal germinal matrix hemorrhage. The incidence of germinal matrix hemorrhage was 3.1% (one of 32) among infants born to women with toxemia, and 23% (55 of 240) among those born to women without toxemia. The apparent protective effect of toxemia could not be explained by intrauterine growth retardation, mode of delivery, or maternal receipt of any medication. Infants born to toxemic women were less likely than their peers to develop pneumothorax, become acidotic, and to require extensive respiratory assistance. This apparently protective effect of maternal toxemia was not seen in infants born to nontoxemic, hypertensive women. Thus, maternal toxemia, but not hypertension, might reduce the risk of germinal matrix hemorrhage by reducing the occurrence and/or severity of pulmonary and related problems that place infants at high risk of germinal matrix hemorrhage. PMID- 3047607 TI - Clinical and morphologic aspects of the vanishing twin phenomenon. AB - The pathologic findings in placentas from ten multiple gestations complicated by the so-called vanishing twin phenomenon were studied to confirm the ultrasonographic evidence. Five pregnancies resulted from in vitro fertilization and embryo transfer, and five conceptions were spontaneous. The pregnancies were studied by repeat ultrasound examinations between five and 12 weeks' gestation. First-trimester bleeding was the only clinical sign of this phenomenon. Postpartum evidence of the vanishing twin phenomenon was found in five cases. Morphologically, the lesions were characterized by well-delineated plaques of perivillous fibrin deposition, associated in one case with embryonic remnants. This focal degenerative change of the placental mass, which also exists in about 25% of placentas from uncomplicated term pregnancies, may be the only clue to the disappearance of one conceptus. PMID- 3047608 TI - Prenatal diagnosis of cysts of the fetal choroid plexus. AB - Over a two-year period, cysts of the fetal choroid plexus were diagnosed prospectively by routine second-trimester ultrasonography in five patients, representing 0.18% of the population scanned for standard obstetric indications. Gestational age at the time of diagnosis ranged from 16-22 weeks. The cysts were located in the posterior portion of the choroid plexus within the lateral ventricle. The maximum diameter ranged from 3-14 mm. In two cases, the cyst was noted to be bilocular. No additional anomalies were detected in any fetus. Follow up sonography two to five weeks after the initial scan documented disappearance of the cysts in all cases. The course of pregnancy in these patients was otherwise uneventful, and all infants were normal physically and neurologically both at the time of birth and between four and 24 months of follow-up. PMID- 3047609 TI - Sonographic evaluation of the normal developmental anatomy of fetal cerebral ventricles: I. The frontal horn. AB - A prospective ultrasound study was conducted in 179 normal pregnant women with gestational ages ranging from 15-40 weeks. Several biometric measurements were obtained throughout pregnancy, including the cerebrofrontal horn distance of the lateral ventricle, the frontal hemispheric width, and the calculated ratio of cerebrofrontal horn distance/hemispheric width. Curvilinear relationships were found between cerebrofrontal horn distance and gestational age (R2 = 0.597; P less than .0001) and between cerebrofrontal horn distance and the biparietal diameter (R2 = 0.618; P less than .0001). In addition, a curvilinear relationship existed between cerebrofrontal horn distance/hemispheric width ratio and gestational age (R2 = 0.492; P less than .0001) and biparietal diameter (R2 = 0.930; P less than .0001). These data represent a comprehensive characterization of normal growth of the fetal frontal horns. They provide a method by which variations from the norm can be assessed and early prenatal diagnosis of developmental anomalies of the fetal ventricular system can be made. PMID- 3047610 TI - Comparison of continuous-wave and pulsed Doppler S/D ratios of umbilical and uterine arteries. AB - Continuous-wave and pulsed Doppler ultrasound measurements of blood flow velocity waveforms of both uterine and umbilical arteries were performed on 85 patients to assess the correlation between these two measurements techniques. Peak systolic to end-diastolic (S/D) ratios measured by continuous-wave devices were not statistically different (P greater than .05) from those measured by a pulsed Doppler device. The S/D ratios of the umbilical artery measured by each device showed a strong correlation, whether measured by one observer (r = 0.93) or two observers (r = 0.89). Uterine artery ratios showed a weaker, although still significant, correlation (r = 0.57). Pulsed Doppler is not routinely necessary for the identification and measurement of umbilical and uterine artery S/D ratios. PMID- 3047611 TI - Fetal umbilical velocimetry using continuous-wave and pulsed-wave Doppler ultrasound in high-risk pregnancies: a comparison of systolic to diastolic ratios. AB - Systolic to diastolic (S/D) ratios of umbilical velocimetry using either continuous-wave or pulsed-wave Doppler ultrasound have been used to assess downstream placental resistance. The purpose of this study was to compare the S/D ratios obtained by both types of instrumentation to determine whether there are significant differences between measurement values. Umbilical velocimetry was performed on 200 high-risk pregnancies in the third trimester using the Angioscan III to obtain continuous-wave velocimetry and the General Electric RT3600 to obtain pulsed-wave velocimetry. Systolic to diastolic ratios were considered abnormally high if they were greater than 3. One hundred sixty-five patients had normal S/D ratios and 35 patients had elevated ratios on both continuous-wave and pulsed-wave ultrasound. There was no significant difference in the mean S/D ratios obtained by either method for the entire population (continuous-wave S/D 2.81 +/- 1.79, pulsed-wave S/D 2.71 +/- 1.83, R = 0.98), the normal group (continuous-wave S/D 1.96 +/- 0.41, pulsed-wave S/D 1.95 +/- 0.40, R = 0.91), and the abnormal group (continuous-wave S/D 6.23 +/- 1.58, pulsed-wave S/D 6.35 +/- 1.52, R = 0.94). Least-square regression was performed to model the relationships between pulsed wave and continuous wave, with both used as dependent variables. The slopes and intercept for the normal and abnormal groups were evaluated and were significantly different (P less than .01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3047612 TI - Intrauterine growth retardation--a prospective study of the diagnostic value of real-time sonography combined with umbilical artery flow velocimetry. AB - This study was undertaken to evaluate the role of umbilical artery flow velocimetry combined with sonographic estimation of fetal weight, head circumference to abdominal circumference ratio, femur length to abdominal circumference ratio, and qualitative determination of amniotic fluid volume as a comprehensive test for the detection of intrauterine growth retardation (IUGR). The following cutoff values were used to indicate abnormal test results: 1) umbilical artery peak systolic to end-diastolic ratio (S/D) above 3, 2) estimated fetal weight below the tenth percentile for gestational age, 3) head circumference to abdominal circumference ratio more than 2 SD above the mean for gestational age, 4) femur length to abdominal circumference ratio above 23.5%, and 5) qualitative amniotic fluid volume less than 2 cm. The study population consisted of 127 patients referred with a clinical suspicion of IUGR. Forty-five infants (35%) were small for gestational age. None of these five tests were uniformly successful in identifying growth-retarded infants. Overall, the best predictor appeared to be estimated fetal weight below the tenth percentile for gestational age, which correctly identified 39 of the 45 IUGR infants (sensitivity 87%, specificity 87%). The sensitivity of this test was nearly twice that of any other test. All indices performed similarly in predicting the non IUGR infant (range of specificities 87-98%). PMID- 3047613 TI - Obstetric characteristics and neonatal performance in a four-year small for gestational age population. AB - Obstetric and neonatal performance were analyzed in an ultrasound-dated small for gestational age (SGA) population from 1982-1985. Eighty-three percent of 160 SGA infants were identified antenatally by means of intrauterine growth retardation (IUGR) risk scoring, and the pregnancies were supervised at a high-risk clinic. Fifty percent were delivered electively, predominantly in gestational weeks 38 39. Thirty percent were born preterm. The cesarean section rate was 40%. Perinatal mortality was 6%, or 4% when lethal malformations were excluded, ten times higher than the corresponding total population figures. Eleven percent of the fetuses had severe malformations. In the remaining SGA population, one infant died after experiencing severe perinatal asphyxia and another developed cerebral palsy; no other major sequelae were found before the age of 18 months. Hypoglycemia and hypothermia occurred frequently, but these problems were managed satisfactorily. The mean hospital stay for term infants was twice that of appropriate for gestational age infants. We conclude that the extra attention paid to the SGA population is well motivated. Future efforts should be directed toward improving the diagnostic techniques for IUGR, as most of the perinatal mortality occurred among SGA infants not identified before birth. PMID- 3047614 TI - Comparison of serum CA 125, clinical impression, and ultrasound in the preoperative evaluation of ovarian masses. AB - The ability to differentiate a malignant from a benign ovarian mass was assessed for four diagnostic procedures: serum CA 125, clinical examination, original ultrasound, and reviewer ultrasound interpretation. When these tests were used individually, the sensitivity and specificity of CA 125 levels were equal to those of a review ultrasound. Overall, the sensitivity of clinical impression and original ultrasound was poor. Sensitivity and specificity were highest for CA 125 assays in postmenopausal patients, especially when these were used as the second diagnostic test. Positive and negative predictive values significantly increased among postmenopausal patients when CA 125 was added to any of the other diagnostic tests examined. In conjunction with such tests, measurement of serum CA 125 significantly increased diagnostic accuracy and may thus have an important role in the preoperative evaluation of women with ovarian masses. PMID- 3047616 TI - Variations in Medicaid surgical rates across counties. An Ohio Department of Human Services Study--1982-1985. PMID- 3047615 TI - Fetal craniofacial morphometrics: in utero evaluation at 16 weeks' gestation. AB - Although ultrasound has proved useful in the diagnosis of fetal craniofacial malformations, its success has been based primarily on subjective clinical observations of apparent abnormal fetal structures, their proportions, and unusual features. However, such clinical observations may be misleading and ideally should be validated by standardized quantitative measurements. We describe here an ultrasonographic methodology that has provided quantitative data describing the normal fetal craniofacies at 16 weeks of gestation. This report is part of an ongoing research project directed at describing fetal facial morphology in utero at different gestational ages. Such data can be used to construct growth curves to which observations from suspected abnormal fetuses can be compared. A total of 53 patients were evaluated at 16 weeks of gestation, at which time 24 craniofacial linear and angular measurements were made. The landmarks employed for these measurements were those used in roentgencephalometry so that this fetal data base could be related to postnatal populations. Such data will contribute not only to a description of facial dysmorphogenesis but also to a better understanding of normal facial growth and development. Furthermore, they constitute a useful tool for prenatal diagnosis, gestational aging, growth predictions, and perhaps for as yet relatively unexplored fields such as fetal therapy. PMID- 3047617 TI - An American College of Surgeons response to the ODHS survey. PMID- 3047618 TI - Implantation of a posterior chamber lens without capsular support during penetrating keratoplasty or as a secondary lens implant. AB - We have devised a technique to fixate a posterior chamber lens in the ciliary sulcus when no posterior capsular support exists. Our short-term follow-up of 22 eyes with lenses thus fixated has shown these eyes to be quiet and to have good pupillary motility after at least 3 months. We believe this new technique is a significant advancement in corneal and anterior segment surgery. PMID- 3047619 TI - [Various characteristics of the surgical treatment of dystrophic varus deformity of the femur neck in children]. PMID- 3047620 TI - [A method of treating open injuries of the calcaneal (Achilles) tendon]. PMID- 3047621 TI - Two cases of left-sided gallbladder associated with cholelithiasis: review of the literature in Japan. PMID- 3047622 TI - [Energy metabolism, food utilization and growth in low birth weight infants]. PMID- 3047623 TI - [Current problems in the treatment of adult acute lymphoid leukemia]. PMID- 3047624 TI - [Incidence of IgA (monomer-dimer, IgA1-IgA2) and IgG producing cells in the tonsils of patients with IgA nephropathy]. PMID- 3047625 TI - [Pathography in medieval legends and canonization documents]. PMID- 3047626 TI - [The last drop in the bucket: Ivanka (a contribution to the pathography of Szchenyi)]. PMID- 3047628 TI - [Nursing education at a university--historical development]. PMID- 3047627 TI - [Birth-bridge--search for identity (a contribution to the pathography of Szechenyi)]. PMID- 3047629 TI - Patterns and contrasts in ophthalmic investigation. AB - The Snellen test has been the most popular clinical measurement of spatial vision for over a century, but it does not fully express the visual ability of an individual. For more analytical purposes the information capacity of the visual system may be assessed by tests of contrast sensitivity and peripheral vision. The visual system selectively reduces the spatial information content of the visual field to avoid overloading the limited capacity for perception and decision making in the brain. The ways in which this reduction occurs and the processing of spatial information is of interest in many disciplines, and theoretical knowledge has been accelerated by the study of artificial intelligence. These processes may be investigated in human subjects by psychophysical and electrophysiological techniques. This provides additional information for diagnostic purposes and will form the basis of new systems of clinical investigation. PMID- 3047630 TI - A method of determining the equivalent powers of the eye and its crystalline lens without resort to phakometry. AB - If the positions of the principal points of the crystalline lens are conjectured, its equivalent power and that of the eye can be calculated as described from ocular dioptrics. The only data required are the subject's refractive error and the results of keratometry and A-scan ultrasonography. The method assumes the profile of the lens to conform to an arbitrary norm. If the unknown lens profile of a given eye differs from this norm, errors will arise. These are shown to be relatively small, and even in extreme cases unlikely to exceed +/- 1.0 D for the lens and +/- 0.5 D for the eye. PMID- 3047631 TI - A century of visual research at Imperial College, 1886 to 1986. PMID- 3047632 TI - The concept of primary fibromyalgia (fibrositis): clinical value, relation and significance to other chronic musculoskeletal pain syndromes. AB - PFS, MPS and TMPDS can be identified using positive diagnostic criteria among patients presenting with chronic pain. A directed rather than exhaustive search for organic diseases known to coexist with these syndromes is usually all that is necessary. Criteria are presently empirical but do identify homogeneous populations of patients for study and treatment. Some patients, however, provide examples of overlap and it may be useful to think of CMPS in terms of the Venn diagram depicted in Fig. 2. In this report we have attempted an initial classification for a group of common and perplexing chronic pain disorders of the musculoskeletal system which at present have no identifiable cause. Previous investigations have been hampered by erroneous pathological concepts, heterogeneous patient populations and poor study design particularly with respect to treatment modalities. We hope that this classification, while empirical, will lead to needed epidemiological studies outlining the similarities and differences between these clinically observable and different musculoskeletal syndromes. We hope, further, that it will foster cooperation between different medical disciplines so that clinical biases might be tested in light of current concepts of the scientific method. PMID- 3047633 TI - Islet-cell antibodies detection using porcine and human pancreas in type I insulin-dependent diabetes (IDD). PMID- 3047634 TI - Physical and social condition of rehabilitated spinal cord injury patients in Japan: a long-term review. AB - In 1983, 28 Rohsai Hospitals in Japan cooperated to study 926 spinal cord injury (SCI) patients to reveal the problems of their rehabilitation. Fifty per cent complained of poor physical condition and were anxious about their health. In addition to complications rising from the SCI, the morbidities of heart disease, diabetes mellitus, liver disease, hypertension and CVA were higher than the Japanese average. It was noted that 1) 44% of tetraplegic patients were confined to living in their home. 2) Ageing exerted a serious influence upon daily life. 3) Crutch gait for patients with paraplegia was not practical. It was also shown that utilisation of automobiles played an important role in extending social activities. For SCI patients, especially those with tetraplegia, it was very difficult to find employment. The rate of employment was only 30% in all and 46% of these were self-employed. PMID- 3047636 TI - [Cancer: what choices for research?]. PMID- 3047635 TI - Measurement of residual urine volume by means of ultrasonic scanning: a comparative study. AB - The basis for this paper was 107 ultrasonic examinations of the bladder in 20 patients with neurogenic bladder dysfunction. The residual urine volume was estimated in three different ways. 1. The residual urine volume is approximately equal to the product of the width, height and depth of the bladder multiplied by a correction factor. 2. By a new method: The residual volume is approximately equal to the cross-sectional area of the bladder when measured in the sagittal plane multiplied by the width of the bladder. 3. By the nomogram method: The cross-sectional area in the sagittal and the transverse plane are respectively plotted into a specially designed nomogram giving an estimate of the residual urine volume. The bladder was emptied by catheterisation immediately after the ultrasonic examinations. Of the three methods, number 2 was found to give the most accurate estimate of residual urine volume. PMID- 3047637 TI - Comparative biochemical and morphometric studies on corneal wound healing. AB - Comparative biochemical and morphological studies were carried out on wounded corneas in order to correlate biochemical findings with morphometric observations during the healing process. Experimental production of corneal wounds, biochemical determinations and quantitative morphometric studies are described in an attempt to correlate corneal matrix macromolecules biosynthesis during the healing process with the morphological modifications of the tissue. The central part and the peripheral part of the corneas were examined separately and compared to the central and peripheral parts of the controlateral corneas. The DNA content of the central portion of the wounded corneas progressively increased and reached after 60 days the same level as the corresponding portion of the controlateral corneas. The DNA contents of the peripheral portions of wounded and controlateral corneas are persistently higher than the DNA contents in the central portion. In peripheral portions of wounded corneas the DNA content is higher than in controlateral corneas and continues to increase steadily during the 60 days of experimentation. Cell density, as determined by morphometry, increased also during that period. The differences between the evolution of the DNA contents and cell density curves may be attributed to variations in cell sizes. The collagen content, estimated from hydroxyproline determinations remained lower in the wounded corneas as compared to the controlateral corneas, even 60 days after operation. This was true for the central as well as for the peripheral portions, suggesting a collagenolytic process as a result of wounding. This is confirmed by morphometric evaluation of fiber density.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3047639 TI - [Habekacin: a new aminoglycoside. Study of nephrotoxicity in rats in comparison with gentamicin, netilmicin and amikacin]. AB - Habekacin is a new aminoglycoside antibiotic. In this study we want to know the effect of increasing dose of habekacin on renal function and on renal morphology. We decide to compare the renal alterations induced by habekacin to these provoked by gentamicin, netilmicin and amikacin. Female Wistar rats received intraperitonally a single injection daily of 10, 30, 50, 150 mg/kg of habekacin for seven days. Wistar rats received also 50 mg/kg gentamicin, 50 mg/kg netilmicin and 150 mg/kg amikacin. No mortality was observed in groups treated with 10, 30, 50 mg/kg habekacin but 50 per cent of rats died with 150 mg/kg habekacin. Habekacin--30 mg/kg seven days--induced a decrease of cortical enzymatic activities, an increase of the number of lysosomes, a great accumulation of myeloid bodies, an alteration of lysosomal membranes Habekacin- 50 mg/kg seven days and 150 mg/kg--induced a decrease of creatinine clearance and ultrastructural alterations of renal tubular cells. Comparative studies with other aminoglycosides showed that amikacin--150 mg/kg was the lesser nephrotoxic drug. With a same dose of 50 mg/kg, gentamicin appeared lesser nephrotoxic than habekacin and habekacin seemed to induce a same degree of renal modifications than netilmicin. With the dose of 150 mg/kg habekacin this drug was higher nephrotoxic than 50 mg/kg gentamicin. In conclusion, if it could be necessary to use habekacin and to prefer this aminoglycoside to gentamicin from an antibacterial activity point of view it is necessary to keep in mind that this drug is potentially nephrotoxic and that the dosage had to be strictly respected.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3047638 TI - [Polymorphism of C4 and factor B in type I diabetes]. AB - The haplotypic frequencies of the fourth component of complement (C4) and factor B (Bf) have been determined in 44 Alsatian type 1 diabetics. An increased frequency of the rare allele Bf F1 (9.1% vs 1.5%) and of the silent alleles of C4 (C4 AQO: 21.6% vers 15.5% -C4 BQO: 29.6% vs 16.0%) was observed in diabetics in comparison to the general population of the same geographic area. A complete HLA haplotype determination has been obtained in 24 type 1 French diabetics. Three haplotypes were associated with the diabetic susceptibility: HLA-A30 CW5 B18 BfF1 C4A3BQO DR3 (18.75% vs 0.86%), HLA-A1 CW7 B8 BfS C4AQOB1 DR3 (15.58% vs 4.17%), HLA-A2 CW3 BW62 BfS C4A3B3 DR4 (6.25% vs 0.45%). The authors suggest that the silent alleles of C4 could modulate the expression of the diabetic susceptibility genes by lowering of the serum C4 hemolytic activity. PMID- 3047640 TI - [Study of specific immunoglobulins (total, IgG, IgM, IgA, IgE) in indirect immunofluorescence and ELISA in 40 patients with trichinosis followed over a 9 month period]. AB - The evolution of the classes of specific immunoglobulins (IgG, IgM, IgA, IgE) of 40 cases of human trichinosis was followed over a nine month period by means of an indirect immunofluorescence test (IF) and ELISA. Specific IgG and IgM appear between 4 and 6 weeks after infection and were still demonstrable after 9 months (respectively in 87.5% and 38.5% of the patients with IF, in 85% and 87.5% with ELISA). Specific IgA were mainly seen on the 2nd month and specific IgE were detected in 37.5% of the patients on the 4th month. In this outbreak, specific antibodies seem to appear later than in a second outbreak observed a few weeks later. PMID- 3047641 TI - [Bacterial contamination of the inspiratory circuit of artificial ventilation apparatus: influence of the frequency of circuit renewal and the duration of ventilation]. AB - In a study concerning contamination of artificial ventilators (Drager model UV1), the influence of 2 parameters was assessed: the frequency of changing the circuit (2 and 4 days) and the interval between admission of the patient into the intensive care unit and obtaining the sample. As a function of these variables, 4 groups of 15 patients each were constituted. The levels of contamination noted at 4 sites in the inspiratory phase tubing (cascade humidifier, condensate collector, tubing nearest to the patient and tubing nearest to the humidifier) and in the gas flow showed no significant difference between the groups, regardless of whether the circuit was changed after 2 or 4 days, or whether the patient had been recently admitted to the department or had been there for at least 6 days. Quantitative and qualitative study of bacteria showed that the one(s) contaminating the inspiratory phase tubing were the same as the one(s) colonizing the tracheal secretions of the patient, and that the most contaminated areas were those nearest to the patient (proximal tubing, collector), which confirms the retrograde contamination of the circuit. PMID- 3047642 TI - [Cerebrospinal fluid pleiocytosis in multiple sclerosis]. AB - Comparative study 100 MS patients with CSF pleiocytosis (greater than 2 cell/mm3) and 100 MS patients with normal cell count in CSF (less than 2 cells/mm3), showed that the pleiocytosis is not a severity index. Pleiocytosis was observed in recent MS disease occurring in young adult patients. PMID- 3047643 TI - [Familial deficiency of complement factor 7: association with bacterial meningitis. Apropos of 3 recent cases]. AB - Three recent cases of inherited deficiency of the seventh component of complement (C7) associated with recurrent infectious meningitis are described. Two cases were associated with meningococcal meningitis, the third is the first case report of C7 inherited deficiency associated with Haemophilus parainfluenzae meningitis. Family studies are consistent with inheritance and non-HLA-linked, autosomal codominant trait of the C7 deficiency. The three patients have remained well, following antibiotic treatment. PMID- 3047644 TI - [Recent data on the mechanism of action of theophylline]. AB - Since Sutherland's work simply based on the inhibition of phosphodiesterase was questioned, other researches have been carried out, indicating that many mechanisms located in the nervous system, the smooth muscle, and the immunity system interfere. The participation of receptors to adenosine, intracellular calcium ions, regulating calciproteins and autacoids such as prostaglandins was the purpose of numerous experimental researches that allow today to fill up the gap left by the fact that the inhibitor of phosphodiesterase was questioned. However, there are not sufficient but there are many seducing research and hypotheses. PMID- 3047645 TI - [Action of testicular hyaluronidase on macromolecules of the cutaneous extracellular matrix. Study by computerized image analysis]. AB - The aim of this work was to precise the action of testicular hyaluronidase (Thiomucase, Millot Solac, Sanofi, France) on the extracellular matrix macromolecules of the skin in an experimental animal model. Intradermal injection of hyaluronidase in the rabbit acts on proteoglycans which are degraded. Collagen bundles are dissociated. This is due to endoglycosaminidase activity of the hyaluronidase on dermal proteoglycans. The network of the elastic fibers was not altered by the enzyme injections, this suggest the absence of elastolytic activity in the Thiomucase preparations. The dissociation of collagen bundles and the degradation of proteoglycans are followed by the resynthesis of the initially degraded proteoglycans. The observed morphological changes which follow the intradermal injection of hyaluronidase, confirm the ability of this enzyme to influence the composition and the structure of the dermis. PMID- 3047646 TI - [Receptors of steroid hormones in the kidney]. AB - Several steroid hormones act in the kidney. We have examined, by autoradiography, the precise distribution of receptors for aldosterone, glucocorticoids, vitamin D (1-25(OH)2D3) and estrogens, in the different epithelia of the nephron isolated by microdissection. Specific nuclear binding sites are localized in the distal parts of the nephron, with some variations according to the steroid hormone considered: target cells for aldosterone are located in the distal tubule and cortical collecting duct, glucocorticoid receptors are present in all distal segments, whereas those of 1-25(OH)2D3 are restricted to the thick ascending limb of Henle's loop and to the medullary collecting tubule. Thus, it appears that several receptors coexist in some cell types. No specific nuclear binding sites for estrogens could be detected along the nephron. On the other hand, a non nuclear specific binding for glucocorticoids was observed in the proximal tubule, where specific glucocorticoid effects have been described. By autoradiography on intact target cells, it appeared that aldosterone receptors are essentially (or exclusively) located in nuclei, as was recently described for other steroid hormones. Binding sites for aldosterone are already present in its target cells in the fetal kidney before their functional differentiation. Aldosterone is weakly metabolized in the kidney, without specific tubular localization. It is possible to show some modifications of aldosterone binding sites, at the level of its target cells, in some pathological states, such as hypertension. PMID- 3047647 TI - [Langerhans cell: CD1 antigens and the Birbeck granule]. AB - Langerhans cells are characterized by two types of markers: an ultrastructural marker, the Birbeck granule and different membrane markers: HLA-D antigens, T4 antigen, and some of the CD1 antigens. These antigens which are specific for the epidermal Langerhans cells, are not expressed by the other epidermal cells. Three CD1 antigens are biochemically defined on human thymocytes, they display a glycoprotein chain non covalently attached to beta-2-microglobulin. Only two of these glycoproteins: the T6 and M241 molecules have been detected on Langerhans cells. The presence of these CD1 antigens on Langerhans cells enhances their relationships to thymocytes, in contrast, Langerhans cells cannot be any more easily associated with the macrophage-monocyte lineage. The physiology of the ultrastructural marker is not well known. Its membrane origin has been experimentally proved since T6 antigen has been found closely associated to newly formed Birbeck granules. PMID- 3047648 TI - [Adaptation of a diazoreaction method for determining total and conjugated bilirubins on a parallel analyser]. AB - A diazoreaction method of estimation of bilirubin has been adapted on a FP9 parallel analyzer. Biotrol's kit could be used only after modifications of the protocol. The critical study of this adaptation included trials of within run precision (CV 1.93 to 4.42%), total precision (CV 2.59 to 6.66%) as also evaluation of analytical range: the limit of linearity is 340 mumol/l. The evaluation of inaccuracy performed with patient specimens leads to establishment of follow-up norms and interpretation norms of allometry line. Our whole results are in conformity with the performance standards of the protocol for the validation of methods published by the Societe Francaise de Biologie Clinique. Finally the described method is quick acting, reliable and very inexpensive. PMID- 3047649 TI - Epidemiology of the childhood acute leukemias. AB - Epidemiologic studies of the childhood leukemias have provided information relevant to several aspects of the care and follow-up of these children. The observations made regarding in utero radiation and ALL risk have certainly curtailed the use of routine obstetric diagnostic radiographs; observations regarding the association between birth weight, fetal loss, and other gestational events provide added enthusiasm for further research into basic biologic events occurring during fetal development; and the genetic patterns of disease supply critical information for genetic counseling and follow-up of affected patients and families. Additionally, the continued epidemiologic surveillance of children with cancer serves to form the foundation from which we will assess any future changes in childhood cancer incidence or pattern. Although not discussed here, the epidemiology of late effects, including second malignancies, reproductive function, and neuropsychologic functioning will assume a more prominent role as more children survive ALL and move into adulthood. While analytic studies have yet to yield an association as strong as the lung cancer/cigarette association in adults, future research designed to isolate biologically homogeneous disease populations for study may lead us to new and important associations. The continued cooperation of large pediatric oncology groups and private physicians is crucial as these future investigations are undertaken. PMID- 3047650 TI - Molecular biology of the leukemias. AB - In this article we reviewed some of the important concepts and techniques of molecular biology applied to the study of lymphoid malignancies. Molecular biology is a tool for addressing certain basic research and clinical questions. It is also a key to the development of a new perspective in biology and medicine. For the first time in history the scope of our understanding of health and disease extends from populations at risk, through individual patients, these patients' tumors, the aberrant cells, the altered proteins, deregulated mRNAs, and disrupted chromosomes, down to the ultimate mutated nucleotide which leads to malignant transformation. This conceptual framework is new to our time and makes us participants as physicians, scientists, and members of society in a uniquely exciting era. PMID- 3047651 TI - The cytogenetics of childhood leukemia: clinical and biologic implications. AB - Cytogenetic studies have revealed a broad spectrum of abnormalities in the chromosomal make-up of human leukemic cells. These abnormalities are acquired during the process of malignant transformation within the neoplastic clone and reflect the genetic lesions and ablations that have occurred. Because cytogenetic abnormalities are tightly linked to the molecular events that lead to leukemogenesis, it is not surprising that these features correlate with immunophenotypic and morphologic features of the leukemic cells, as well as with the clinical characteristics of children at diagnosis and their responsiveness to therapy. Molecular analysis of the disordered structure or disrupted regulation of genes located at critical chromosomal breakpoints in leukemic cells should continue to provide important insight into normal and aberrant hematopoietic cell function. PMID- 3047653 TI - Chronic leukemias of childhood. AB - Chronic leukemias account for fewer than 5 per cent of childhood hematologic malignancies. The various subtypes are chronic mylocytic leukemia (adult, juvenile, and familial), chronic myelomonocytic leukemia chronic monocytic leukemia, and chronic lymphocytic leukemia. The most common of these, adult-type chronic myelocytic leukemia, is characterized by specific cytogenetic alterations; recent advances in molecular biology are linking these genetic events to the pathophysiology and course of this fascinating neoplasm. PMID- 3047652 TI - Biologic characteristics and treatment of acute nonlymphocytic leukemia in children. Report of the ANLL Strategy Group of the Childrens Cancer Study Group. AB - Today approximately 75 per cent of children with ANLL can be induced into a complete remission and approximately 40 per cent will have an event-free survival for more than 3 years, irrespective of whether they received a bone marrow transplantation or chemotherapy after induction. In order to achieve these results very intensive therapy is required. The morbidity and mortality of treatment are high. The length of therapy needed after induction of remission is not known. Whether or not maintenance therapy is required is perhaps related most directly to the intensity of the therapy employed. Similarly, the role of bone marrow transplantation in patients in first remission, treatment of CNS leukemia, and treatment of chloromas are controversial. There is general agreement that WBCs over 100,000, acute monoblastic leukemia in infants less than 2 years of age, and certain chromosomal abnormalities are associated with a poor prognosis. Although there has been a dramatic improvement in the treatment of ANLL over the past 15 years, stratification of therapy based on biologic parameters, and alteration of treatment based on the early responses to treatment may be required before further advances will be made. PMID- 3047654 TI - Central nervous system leukemia. AB - With one exception, the risk and severity of neurotoxicity is directly proportional to the number of therapeutic modalities used. Three are worse than two, and two are worse than one. Combinations of therapeutic modalities which include CNS RT appear to be the most neurotoxic. The least neurotoxic combination of two modalities appears to be the IT MTX with high-dose intravenous MTX. Thus far, high-dose MTX appear to be the safest single modality, in terms of acute, subacute, and delayed neurotoxicity. The improved outcome in intellectual and academic performance in the NCI-191/CCG-134P conjoint trial of the CCSG and the Pediatric Branch described above (see section of Presymptomatic CNS Therapy) appears to confirm this observation. Whether triple IT chemotherapy will have the same result remains to be established. If CNS RT must be combined with MTX therapy, the least neurotoxic approach appears to be to administer these modalities in sequence, IT MTX, or high-dose intravenous MTX followed by CNS RT. MTX given during or after CNS RT appears from the clinical data to be more likely to produce severe neurologic sequelae. An ultimate goal would be to avoid both ionizing RT and IT chemotherapy. To this end, the NCI/CCSG study has demonstrated that this may be possible, except for those patients who are at the highest risk for CNS relapse despite conventional CNS prophylaxis. Meanwhile, for presymptomatic therapy, either cranial RT (18 Gy total dose at 120-180 cGy per day) in conjunction with IT MTX, or frequent IT chemotherapy with MTX, cytarabine, and hydrocortisone combined and administered throughout induction, consolidation, and maintenance is eminently justified in the majority of children with ALL. On a worldwide basis, chemoradiotherapy with cranial RT and IT MTX remains the established method of preventing overt CNS leukemia. The benefits of this intervention, in terms of prevention of symptomatic CNS leukemia, prolongation of complete remission, and increased cure rates, are clearly worth the risks. PMID- 3047655 TI - Late effects of antileukemic treatment. AB - Increasing numbers of childhood ALL survivors have increased the need to assess the physical and psychosocial functioning of this group in a careful manner. This article reviews data on the frequency and types of second malignancies, structural and functional changes in the central nervous system, endocrine effects on growth and reproduction, and psychosocial aspects of development. Most long-term survivors of ALL do not have serious or life-threatening medical problems; however, medical and psychosocial problems may not be insignificant and may require coordinated management over prolonged periods. PMID- 3047656 TI - Pharmacologic considerations in the treatment of acute lymphoblastic leukemia. AB - Great strides have been made in the chemotherapy of childhood ALL over the past three decades. Principles and concepts learned from early successes in treating this disease subsequently have been applied to the treatment of other pediatric cancers and adult malignancies. Because of the prominent role of chemotherapy in the treatment of ALL, a clear understanding of the clinical pharmacology of the antileukemic drugs is essential. This article reviews issues relating to the clinical pharmacology of the anticancer drugs that are commonly used in the treatment of acute lymphoblastic leukemia. PMID- 3047657 TI - Preleukemic syndromes and other syndromes predisposing to leukemia. AB - The myelodysplastic and myeloproliferative syndromes are syndromes in childhood that may precede leukemia. Clinical and biologic features are reviewed in this article. Although rare, they offer an unique opportunity to observe the evolution of leukemia and give clues that are helping us to understand the leukemogenic process. PMID- 3047658 TI - Infectious complications in childhood acute leukemias. AB - Infectious complications in children with acute leukemias are reviewed as to incidence, predisposing factors, microbiologic etiologies and treatment. Principles of antimicrobiologic therapy are presented for bacterial, fungal, viral, and protozoal infections seen in children with cancer. Prevention of infection is also discussed. PMID- 3047659 TI - Bone marrow transplantation for treatment of leukemia in children. AB - BMT is a well-established treatment for children with ALL in second remission, ANLL in first and second remission and children with JCML and CML. Improvements in transplantation technology and supportive care have resulted in significant increases in the percentage of long-term survivors of allogeneic marrow transplantation. Newer strategies, such as partially matched donor, unrelated matched donor, and autologous transplants, are bineg pursued to overcome the histocompatability barrier. The development of more effective antileukemic cytoreductive chemotherapy and radiation therapy regimens and better methods of preventing GVHD are areas in which further improvements are necessary. Newer methods of marrow purging, such as the use of monoclonal antibodies linked to immunotoxins, already are being tested. In addition, the recent development of molecularly cloned hematopoietic growth factors, such as CSFGM, may make it possible to improve marrow recovery and hasten return of normal immunologic function, thereby increasing the overall safety of the transplant procedure. It is hoped that these innovations eventually will increase the overall applicability of BMT and its role in the treatment of leukemia. PMID- 3047660 TI - [Monocyte-macrophage activation syndrome]. AB - The activation of macrophage after stimulations (infectious, immunologic, irritative) is the ability of this cell to destroy intracellular bacteria and tumoral cells. Activated macrophage is characterized by morphological, biological and immunological properties and by its phagocytosis ability. The activation of macrophage can explain most of the signs encountered in several diseases: familial hemophagocytic lymphoreticulosis, accelerated phases of some immunodeficiencies but also protection of the organism against infectious diseases, immunological tolerance and antitumoral immunity. PMID- 3047661 TI - [Dumping syndrome: a case after Nissen's operation]. AB - We report on a case of dumping syndrome 1 month after Nissen fundoplication in a 5 month-old infant presenting with symptoms of late hypoglycemia. Blood glucose levels in pre- and post-prandial periods, oral glucose tolerance test and isotopic gastric emptying study confirmed the diagnosis. All symptoms resolved with diet therapy which could be stopped 8 months later. Dumping syndrome is not a so rare complication of gastric surgery and should be considered in patients with non specific symptoms. PMID- 3047662 TI - [Marden-Walker syndrome. New case and discussion about its role in arthrogryposes]. AB - A new case of Marden-Walker syndrome is reported. The Marden-Walker syndrome is a rare entity associating neonatal arthrogryposis and blepharophimosis with autosomal recessive inheritance. Its place among the various syndromes with neonatal arthrogryposis is discussed. PMID- 3047664 TI - [A method for studying the bone mass: dual-photon densitometry]. AB - The assessment of bone mineral mass during growth, periods diseases or therapy which modify bone mineralization in the child is of great interest. Single-photon absorptiometry carried out on forearm bones informs on the mineral density of the compact bone of the appendicular skeleton. It is easy to perform in children and infants. Dual-photon absorptiometry appears to be the most accurate for measuring bone mineral content in endocrine and metabolic bone disorders. The low dose of radiation and the non-invasive nature of this high-precision method render it well-adapted and suitable for measuring bone mineralization in childhood. PMID- 3047663 TI - [Precocious puberty and ovarian follicular cysts]. AB - Ovarian follicular cysts have been detected by ultrasound in 2 to 8 years old girls with precocious puberty, central puberty (2 cases), transient precocity (1 case) or premature menarche (1 case). Complete regression of the cysts was either spontaneous (1 case) or due to suppressive therapy. The mechanisms of precocious puberty with follicular cysts, dependent or independent of gonadotropins, and their spontaneous evolution guide both the management and the therapy with LH-RH analogues and/or aromatase inhibitors. PMID- 3047665 TI - Etoposide and teniposide. Bioanalysis, metabolism and clinical pharmacokinetics. AB - Etoposide (VP 16-213) and teniposide (VM 26) are semisynthetic epipodophyllotoxin derivatives active against a variety of tumours. The clinical efficacy has led to an increasing interest in these compounds. This review presents information on the mechanism of action, biochemical pharmacology, bioanalysis, metabolism and pharmacokinetics of etoposide and teniposide. PMID- 3047666 TI - Home care agencies and community health services accredited by NLN October 1987. PMID- 3047667 TI - Associate degree education for nursing 1988-89. PMID- 3047668 TI - The treatment of venous thromboembolism. AB - Venous thromboembolism is usually treated with heparin followed by oral anticoagulants for approximately 3 months. The optimal length of heparin therapy is uncertain. There is now good evidence that treatment failures are associated with inadequate heparin effects. Heparin can be administered with similar safety and effectiveness by either intravenous infusion or 2 hourly subcutaneous injection. Oral anticoagulant therapy is effective and safe if given in a dose which prolongs the prothrombin time to an international normalized ratio of 2.0 to 2.5. Thrombolytic therapy is indicated in patients with major pulmonary embolism and in selected patients with recent acute venous thrombosis. Vena caval interruption is usually confined to patients who have contraindications to anticoagulant therapy. Surgical removal of thromboembolic obstruction should be considered in selected patients with thromboembolic pulmonary hypertension. PMID- 3047669 TI - Pharmacology of selective thrombin inhibitors. AB - Thrombin plays a key role in thrombosis and haemostasis, and is selectively inhibited by hirudin and synthetic inhibitors. Hirudin, a polypeptide (molecular weight 7,000 daltons) extracted from medicinal leeches, can now be produced by gene technology. Hirudin binds selectively to thrombin with high affinity and inhibits its enzymatic properties. Besides heparin, hirudin is not inhibited by platelet factor 4; it prolongs in vitro and ex vivo routine blood coagulation assays and prevents thrombosis in a number of animal models without increasing haemorrhagic risk. In humans, hirudin disappears from the blood with a half-life of 1 h, is devoid of undesirable side effects and has been shown to be efficient in the treatment of chronic disseminated intravascular coagulation (DIC). A number of synthetic direct thrombin inhibitors have been described, including benzamidine derivatives which share identical pharmacological properties with hirudin; however their biological half-life after i.v. injection is shorter. Other derivatives (amidino-phenyl-pyruvic acid) have longer half-lives and have been used to treat chronic DIC in man. PMID- 3047670 TI - The origin of the Sternberg cell. AB - The Sternberg cell, the malignant cell of Hodgkin's disease, remains of uncertain nature 80 years after it was first described. Numerous hypotheses have been advanced as to its origin: that it derives from the macrophages, B lymphocyte, T lymphocyte, interdigitating reticulum cell or even from an as yet unidentified cell-line. We here review these various hypotheses, as well as the different techniques used for its study. The morphologic, ultrastructural, histochemical and immunologic studies are discussed in detail. PMID- 3047671 TI - A comprehensive list of chemically synthesized genes. AB - A databank for chemically synthesized genes has been compiled. PMID- 3047672 TI - KRAS codon 12 mutations occur very frequently in pancreatic adenocarcinomas. AB - DNAs from human pancreatic adenocarcinomas were analyzed for the presence of mutations in codons 12, 13 and 61 of the NRAS, KRAS and HRAS gene. Formalin-fixed and paraffin-embedded tissue was used directly in an in vitro amplification reaction to expand the relevant RAS sequences. The mutations were detected by selective hybridization using mutation-specific synthetic oligonucleotides. In 28 of the 30 patients we found a mutation in codon 12 of the KRAS gene. This result confirms the findings of Almoguera et al. [Cell 53 (1988) 549-554] that KRAS mutations occur frequently in adenocarcinomas of the exocrine pancreas. The mutations are predominantly G-T transversions, in contrast to the KRAS mutations in colon tumors which are mainly G-A transitions. Furthermore, in a portion of the tumors the mutation appears to be homozygous. PMID- 3047673 TI - Mismatch-containing oligonucleotide duplexes bound by the E. coli mutS-encoded protein. AB - The binding of the mutS gene product, a protein involved in at least two E. coli mismatch correction pathways, to a series of synthetic DNA duplexes containing mismatches or mismatch analogues of the purine/pyrimidine type was studied in order to establish whether a correlation exists between the recognition of these mispairs and the efficiency of their correction in vivo. Experiments using nitrocellulose filter binding or band-shift assays revealed that duplexes containing a G/T mismatch or its analogues I/T and DI/T were bound by the protein with affinities correlating to the efficiency of their repair in vivo. In contrast, the A/C mismatch, contained within the same sequence, was bound only poorly, despite being efficiently corrected in vivo. The analogues of the A/C mispair, uncorrected in vivo, were not detectably bound under the conditions of these assays. PMID- 3047674 TI - The tRNAGlu2 gene in the rrnB operon of E. coli is a prerequisite for correct RNase III processing in vitro. AB - RNase III cleaves precursor 16S RNA and precursor 23S RNA from the ribosomal RNA transcript. In vitro transcription experiments, using plasmids with the rrnB operon truncated in the 16S RNA and with various deletions in the spacer tRNA region, showed that no matter what size of deletion if the tRNA gene was affected RNase III processing of 16S RNA became incomplete. In comparison to a control plasmid, where only the 16S RNA gene was truncated and that showed normal RNA processing, plasmids where the tRNA gene was deleted partially or totally either the 5' or the 3' end of 16S RNA was processed. This relation between RNase III processing and the 3-dimensional structure of tRNA suggests an interaction between RNase III and a tRNA processing enzyme most probably RNase P. PMID- 3047675 TI - Mild temperature shock affects transcription of yeast ribosomal protein genes as well as the stability of their mRNAs. AB - Shifting the temperature of a yeast culture from 23 degrees to 36 degrees C results in a sudden and severe (greater than 85%) decline in the cellular levels of ribosomal protein (rp-)mRNAs. Recovery during continued growth at 36 degrees C occurs within 1 h. The use of hybrid genes carrying different portions of the region upstream of the gene coding for ribosomal protein L25 revealed that this characteristic, coordinate temperature shock phenomenon does not depend on the presence of specific upstream DNA sequences. Analysis of a heterologous gene carrying a synthetic UASrpg (upstream activation site of yeast ribosomal protein genes) provided conclusive evidence that the rp-characteristic, transient heat shock response is not mediated through the UASrpg elements. The addition of the transcription inhibitor 1,10-phenantroline prior to a 23 degrees to 36 degrees C heat shock inhibited the severe decline of the rp-mRNA levels. The latter observation indicates that transcription is required for the rp-gene- specific response to heat shock. A milder temperature shift, from 23 degrees to 30 degrees C, gave rise to a two-fold decrease in mRNA levels for all genes studied, both ribosomal and non-ribosomal. Together, these results indicate that a temperature shift causes a temporary general transcriptional arrest in yeast cells, resulting in an over-all decrease in mRNA levels. In addition, an enhanced nucleolytic break-down of pre-existing rp-mRNAs accounts for the dramatic drop in the steady state amounts of these mRNAs observed upon a 23 degrees----36 degrees C shift. This enhanced breakdown is caused directly or indirectly by a factor whose synthesis is induced by the heat shock treatment. PMID- 3047676 TI - The organization and transcription of the galactose gene cluster of Kluyveromyces lactis. AB - The yeast Kluyveromyces lactis grows on galactose by inducing the Leloir pathway enzymes-kinase, epimerase, and transferase. To investigate the molecular mechanism for regulating expression of this metabolic pathway we isolated GAL1, GAL7, GAL10, which code for kinase, transferase, and epimerase, respectively, and characterized their size, organization, and transcriptional regulation. Our results indicate that induction of the Leloir pathway in K. lactis occurs at the level of transcription and that the organization and regulation of the GAL gene cluster in K. lactis is closely related to the homologous gene cluster in Saccharomyces cerevisiae. Likewise, the Upstream Activator Sequences that regulate induction of the GAL genes are similar in base sequence, number and relative location in the two yeasts. PMID- 3047677 TI - Mutations within the decoding site of Escherichia coli 16S rRNA: growth rate impairment, lethality and intragenic suppression. AB - Several C----U transitions and small deletions were introduced into the conserved region centered on base C1400 in Escherichia coli 16S rRNA by in vitro mutagenesis. The mutations were placed within rrnB operons on multicopy plasmids under the transcriptional regulation of either the normal rrnB P1P2 promoters or the temperature-inducible PL promoter from bacteriophage lambda and introduced into E. coli hosts. When expressed from the P1P2 promoters, several of the mutant 16S rRNAs impaired cell growth while others, including one in which U replaced C at position 1400 within the ribosomal decoding site, had little or no effect on cell doubling time. However, C----U transitions at positions 1395 and 1407, as well as the deletion of C1400, appeared to render their hosts inviable. Cells in which these mutations were expressed from the lambdaPL promoter died within four generations after induction. Unexpectedly, the lethal phenotype was suppressed intragenically by replacement of G1505 with A, C or U. Suppression may alleviate a functional defect in 30S subunits containing the U1395, U1407 or deltaC1400 mutations. PMID- 3047679 TI - A procedure for in vitro amplification of DNA segments that lie outside the boundaries of known sequences. PMID- 3047678 TI - 60Co gamma-rays induce predominantly C/G to G/C transversions in double-stranded M13 DNA. AB - Upon irradiation with gamma rays of an oxygenated aqueous solution of double stranded M13 DNA, a very specific mutation spectrum was found with respect to both the type and the positions in the DNA sequence. Of the 23 mutations, which were sequenced, 16 represent a C/G to G/C transversion. A C/G to T/A transition was found once and a G/C to T/A transversion twice. The remaining 4 mutations are frameshifts, 2 are identical and formed by the insertion of a G/C basepair; the other 2 mutations are due to a duplication of 10 basepairs situated at different positions but with a remarkable homology in base sequence. Fourteen mutations, including the 2 duplications are found in the neighbourhood of a TGCT/ACGA sequence. PMID- 3047680 TI - The UAS of the yeast PGK gene is composed of multiple functional elements. AB - The UAS (upstream activator sequence) of the yeast PGK gene contains a transcriptional activator domain located between bases -479 and -402 upstream from the initiating ATG. This region of the UAS contains three direct repeats of the sequence 5'CTTCC3'. The roles in transcriptional activation of these repeats and other sequences within the activator domain were investigated. When short regions containing the repeats were removed PGK expression was considerably reduced, the magnitude of the effect depended upon which CTTCC block was absent. Sequences between -473 and -458 which did not contain a CTTCC block were also shown to be necessary for high levels of PGK expression. A DNA fragment containing activator sequences up to -473 was shown to interact specifically in vitro with a yeast nuclear protein extract. DNase I footprinting identified a protected region between -473 and -458 and single base changes in DNase I sensitivity at the CTTCC repeats. PMID- 3047681 TI - Oligonucleotide site-directed mutagenesis in Xenopus egg extracts. AB - Addition of M13mp18 single-stranded DNA annealed with an oligonucleotide to a Xenopus egg extract results in a rapid and efficient incorporation of the oligonucleotide in a complete double-stranded supercoiled molecule. Both the efficiency of DNA synthesis and the recovery of complete double-stranded molecules are increased relative to the reaction carried out by the classical technique using the E. coli Klenow DNA polymerase, DNA ligase, dNTPs, ATP and ions. Site specific mutagenesis was assayed by reverting a point mutation in the lacz region of M13mp18. The color assay described by Messing and sequencing of the DNA extracted from isolated plaques was used to check for the reversion. A 2 hr incubation of the heteroduplex carrying the mutagenic oligonucleotide in the Klenow-ligase-dNTP mixture allows a recovery of 6% mutant phage after transformation of competent cells with the reaction products. Using the Xenopus egg extract, 83% mutant phage were recovered after the same incubation time, in reactions entirely performed in parallel. The Xenopus extract is stable and contains all components required for the assay, including all ionic and protein factors; thus the only addition is the annealed DNA. Such an eukaryotic system is therefore an attractive alternative to the reconstituted prokaryotic DNA polymerase-DNA ligase system for site specific mutagenesis. PMID- 3047682 TI - The nucleotide sequence and gene organization of red clover necrotic mosaic virus RNA-2. AB - Red clover necrotic mosaic virus, a member of the dianthovirus group, is characterized by a genome composed of two nonhomologous single-stranded RNAs of approximately 4.0 (RNA-1) and 1.4 kb (RNA-2). The complete nucleotide sequence of the RNA-2 has been determined. RNA-2 is 1448 nucleotides in length with a 5' terminal m7G cap and no 3' terminal poly-A tail or 5' terminal VPg. An open reading frame beginning at the first initiation codon at nucleotide 80 and ending at nucleotide 1030 has been identified which can encode a polypeptide of 35 kDa. RNA-2 directs the synthesis of a 35 kDa polypeptide in vitro. SP6 and T7 transcripts from full length RNA-2 cDNA clones directed the synthesis of a polypeptide with the same electrophoretic mobility as the polypeptide directed from authentic RNA-2. Clones with various 3' terminal deletions both outside and within the 35 kDa open reading frame were transcribed and translated in vitro to define the limits of the 35 kDa open reading frame. A second, small open reading frame capable of encoding a polypeptide of 4.9 kDa was also indicated from the sequence; however, there was no evidence for a protein product of that size. RNA 2 is presumed to be monocistronic and encode a cell-to-cell movement function. A small but significant amino acid sequence homology was observed with the brome mosaic virus RNA-3a polypeptide. PMID- 3047684 TI - The yeast SUF3 frameshift suppressor encodes a mutant glycine tRNA(CCC). PMID- 3047683 TI - The yeast SUF5 frameshift suppressor encodes a mutant glycine tRNA(CCC). PMID- 3047685 TI - Nucleotide sequence of the Escherichia coli purF gene encoding amidophosphoribosyltransferase for de novo purine nucleotide synthesis. PMID- 3047686 TI - Hybridisation detection of single nucleotide changes with enzyme labelled oligonucleotides. PMID- 3047687 TI - Dot blot detection of point mutations with adjacently hybridising synthetic oligonucleotide probes. PMID- 3047689 TI - A new method to obtain high DNA transformation efficiency of E. coli competent cells. PMID- 3047688 TI - pIN-III-ompA secretion vectors: modification of the ompA signal peptide sequence for easier insert cloning. PMID- 3047690 TI - A plasmid vector for cloning directly at the transcription initiation site of a bacteriophage T7 promoter. PMID- 3047691 TI - Influence of a nursing intervention on regimen adherence and societal adjustments postmyocardial infarction. AB - One hundred three first-time myocardial infarction (MI) patients were randomly assigned to either an experimental or a control condition. Patients completed a cardiac rehabilitation program during hospitalization and were interviewed to assess intentions to follow regimen prescriptions, attitudes toward the prescriptions, coping methods, and the perceived beliefs of others concerning their intentions. Patients were visited at home 30 days after discharge and reassessed on each of the above variables except that their behavior was substituted for intentions and societal adjustment was assessed. The experimental group was given an intervention program which included a discussion of assessment data, identification of problems, and establishment of goals. The assessment was repeated 60 days after discharge. No differences were found between experimental and control groups for either medical regimen adherence or societal adjustment. There was a significant decrease in mean scores for all variables from hospital to 30 days for both groups, but no change from 30 to 60 days. Attitudes and perceived beliefs of others were predictive of adherence, and it was concluded that these variables need to be included in any rehabilitation program. PMID- 3047692 TI - Nursing interventions and patient outcomes: a meta-analysis of studies. AB - The 84 subject-studies and 4,146 individual subjects in this meta-analysis were obtained from nurse-conducted experimental research over an 8-year period. The entire universe of accessible subject-studies that met criteria was included. Although both published and unpublished research were included to protect the study from publication bias, there was no statistically significant difference in findings. The mean effect size for the sample of comparisons from the 84 studies was .59. The associated U3 value of 72.2 and r of .28 indicate that patients who receive research-based nursing interventions can expect 28% better outcomes than 72% of the patients who receive standard nursing care. PMID- 3047693 TI - Re: 'Outcome of multiple usage of disposable syringes in the insulin-requiring diabetic'. PMID- 3047694 TI - Prevailing over pain. PMID- 3047695 TI - Nursing: the hospital's competitive edge. AB - The health care marketplace is becoming increasingly competitive. The hospital has a built-in marketing force with the nursing department, because nurses are in constant, direct contact with the customer. Nursing must identify the case mix profile of the community and focus the hospital product lines to meet community needs. The nursing department should decentralize, change, measure, and innovate the staff mix needed to operationalize these product lines. The development of nursing practice standards for the case mix will help to identify the staff mix needed and create systems to efficiently manage the product lines. Nursing management must become aware of cross-subsidization and downward skill substitution of nursing personnel. Nursing information systems must generate quality reports that invoke cost consciousness on the part of nursing staff. Quality assurance programs must become unit based and complete with frequent audits to correlate length of stay with nursing quality. Correlations must be determined between nursing productivity and case mix to determine the hospital's niche in the marketplace. The transformation of health care into a competitive business industry has created many opportunities for nursing. The health care industry's incentives for efficiency along with the decreasing demand for inpatient hospital services will be the forces driving health care toward a competitive marketplace. The hospital's nursing department should be strategically positioned to become accountable for increasing market share and enhancing quality patient outcomes. The focus has shifted from the theoretical to the tactical, which is a step in the right direction, particularly for nursing. Nursing, if strategically positioned, will not only thrive but will also excel in this chaotic environment by capturing the opportunities and being innovative. PMID- 3047696 TI - Quality: the banner of the 1980s. AB - Quality management is the vehicle for integrating the critical aspects of patient care into nursing service delivery approaches. Predetermined standards for quality, productivity, risk, and cost are correlated in this quality management approach. Information systems facilitate this integration. Nurses must now structure their quality management data elements to accurately reflect nursing practice and relationships of nursing care to the whole. PMID- 3047697 TI - Directions and dilemmas in nursing quality assurance. AB - Effects of the health care environment have been a catalyst for the evolution of nursing quality assurance programs. Changes can be summarized as an increased focus on actual clinical practice, an increased participation by professional nurses, and an increased sophistication in the methods used to monitor and evaluate. Much work remains to be done in creating environments that assure quality of care. PMID- 3047698 TI - Production and exocrine secretion of LHRH-like material by the male rat reproductive tract. AB - Luteinizing hormone-releasing hormone (LHRH)-like immunoreactivity was localized in the male reproductive system of the rat. Epithelial cells of the epididymus, seminal vesicles and coagulation gland showed a strong reaction to anti-LHRH serum. Also the epithelia of the ductus deferens and the prostate gland appeared to be immunoreactive, albeit to a lesser extent. The LHRH-like substances are most likely secreted into the male tract, as can be concluded from the observation that the secretion product in the lumina of the seminal vesicles, coagulation gland and prostate gland was also immunopositive. The functional significance of these phenomena is discussed. No immunostaining was obtained with antisera to FSH, LH or beta-hCG. PMID- 3047699 TI - Effect of the secretin family of peptides on gastric emptying and small intestinal transit in rats. AB - We have compared the effects of the secretin family of peptides and their synthetic fragments on gastric emptying (GE) and small intestinal transit (SIT) using an unanesthetized rat model which simultaneously measures the GE and SIT of both solids and liquids. The meal consisting of 5% polyethylene glycol w/v, 5% Indian ink v/v and 20 non-digestible plastic beads was given intragastrically 10 minutes after the intraperitoneal injection of 0.5 ml of saline or peptides (2 and 5 micrograms/kg). Plasma secretin and the immunospecificity of secretin fragments were determined. In control rats, the t1/2 for the GE of both solids and liquids were 56 +/- 3.8 and 19 +/- 2.3 minutes, respectively. Liquids emptied faster than the solids and liquids travelled ahead of the solids in the intestine. Secretin (5 micrograms/kg) inhibited GE of both solids and liquids by 33-37%. Secretin delayed the SIT of the meal by approximately 35%. Fragments of secretin and of VIP had no effect on GE and SIT of both solids and liquids. The whole molecule of secretin was required to inhibit GE and to delay SIT of solids and liquids. Glucagon, PHI and growth hormone releasing factor (GHRF1-44) inhibited GE and SIT of both solids and liquids. For all peptides tested, the inhibition of SIT was proportional to the inhibition of GE suggesting that the prolongation of SIT was secondary to delayed GE. These observations indicate that the peptides of the secretin family inhibit GE and prolong SIT. Thus, the structural requirement required for the secretin family of peptides to effect their motor actions on the stomach is similar to that required for pancreatic enzyme secretion. PMID- 3047701 TI - Anxiety disorders. Relationships to other psychiatric illness. AB - Considerable evidence suggests that patients with anxiety disorders have secondary depression, and somewhat less evidence suggests that secondary alcoholism is seen in anxiety disorder. Such illnesses as manic-depressive disorder, schizophrenia, and hysteria are likely to be associated with secondary anxiety syndromes. Considerable data support the idea that within families of anxiety disorder patients alcoholism is seen more frequently than would be expected by chance and that within families of alcoholics anxiety disorders are seen more frequently than would be expected by chance. These findings suggest a difference in expressivity for the same familial propensity. PMID- 3047700 TI - Differential distribution of two molecular forms of gonadotropin-releasing hormone in discrete brain areas of goldfish (Carassius auratus). AB - Two molecular forms of gonadotropin-releasing hormone (GnRH) were identified in the extracts of various brain areas, spinal cord and pituitary in female and male goldfish and had chromatographic and immunological properties similar to [His5, Trp7, Tyr8]-GnRH (cGnRH-II) and [Trp7,Leu8]-GnRH (sGnRH). Radioimmunoassay using different GnRH antisera after high pressure liquid chromatography did not reveal significant peaks of mammalian GnRH, [Gln8]-GnRH and [Tyr3,Leu5,Glu6,Trp7,Lys8] GnRH in the brain extracts. The proportion of cGnRH-II-like immunoactivity to sGnRH-like immunoactivity was higher in the caudal brain areas compared to the rostral areas. The differential distribution of two GnRH forms suggest that the different GnRH forms may have different physiological functions. PMID- 3047702 TI - Basic and clinical studies with corticotropin-releasing hormone. Implications for a possible role in panic disorder. AB - We have presented several lines of circumstantial evidence suggesting a possible role for CRH in the panic disorder syndrome. Such a role has by no means been demonstrated and is indeed challenged by several lines of contradictory data. Critical evaluation of this hypothesis must await further basic research on the role of hypothalamic and extrahypothalamic CRH in stress-mediated phenomena and on their interrelationships. Moreover, additional clinical research will be required to further delineate the landscape of hypothalamic-pituitary-adrenal function in patients with panic disorder both during acute panic episodes and intercurrently. PMID- 3047703 TI - Similarities in hypothalamic-pituitary-adrenal axis activity between patients with panic disorder and those experiencing external stress. AB - Psychologic stressors are less potent stimuli of the HPA axis in humans than physical stressors, but they can cause mild changes in acute ACTH and cortisol production. These changes, however, are generally not of sufficient magnitude or duration to cause measurable changes in UFC excretion. Furthermore, chronic stress leads to an attenuation of the HPA axis response. This basic knowledge concerning psychologic stress helps explain the reason why patients with uncomplicated anxiety/panic disorder, a condition involving similar symptoms, do not demonstrate UFC abnormalities. Panic disorder, when accompanied by depression, however, is associated with an increase in UFC excretion which is probably more related to the depression than the panic state. Interestingly, panic disorder, when accompanied by agoraphobia, also shows an elevation in UFC levels which makes it endocrinologically distinct from uncomplicated panic disorder. The reason for this is unclear. Treatment of the panic disorder with benzodiazepines does not further lower the UFC levels in patients with uncomplicated panic disorder despite clinical improvement, but it does lower UFC levels into the normal range in those with concurrent depression and agoraphobia. Alteration in the UFC level with treatment is only partially related to clinical improvement. PMID- 3047704 TI - Immune system. Relationship to anxiety disorders. AB - The demonstration that behavioral states and CNS processes are associated with immune function suggests that there may be a relationship between anxiety and the immune system. Stress and immunity have been studied extensively, but there have been relatively few studies of anxiety and immunity. Many of the neurobiologic processes associated with stress and with depression have been observed in anxiety and are known to influence the immune system. A review of the immune response to stress and of immune alterations in depression has been presented in an effort to provide further understanding of the biology of anxiety. It appears that a variety of factors such as age; sex; nature, intensity, and chronicity of a stressful life events; and psychologic response to life stress need to be considered in the investigation of behavior and immunity. The biologic effects of stress on immunity are multifaceted, including complex neuroendocrine and neurotransmitter interactions. Further investigation is required of anxiety and immunity in clearly delineated and diagnosed anxiety states and disorders. Such studies may help to elucidate the pathophysiology of anxiety disorders. PMID- 3047705 TI - Panic disorder and the vestibular system. AB - This article reviews the interrelationship between panic disorder and vestibular function. There is a possibility of both somatopsychic and psychosomatic interactions between panic and the vestibular system. Another possibility is that vestibular dysfunction could be associated with certain mental disorders, including panic disorder, as a nonspecific marker. Somatopsychic interactions are suggested by findings of high prevalence of vestibular dysfunction in selected patients with panic disorder, by the occurrence of "space and motion phobia" in patients with panic disorder, and by the report of anxiety and pseudoagoraphobia in some patients with a primary complaint of vertigo. Psychosomatic influences include symptoms of dizziness and increased sensitivity of the vestibular system due to anxiety or hyperventilation. Vestibular dysfunction as a nonspecific marker is discussed in the context of a review of studies of the vestibular system in schizophrenia. Before more definite conclusions can be drawn whether panic disorder is related to vestibular dysfunction in some cases, further research is needed to establish the specificity of vestibular dysfunction for panic disorder. PMID- 3047706 TI - The peripheral sympathetic nervous system. Its role in normal and pathologic anxiety. AB - Acute emotional arousal, regardless of the emotional state, increases sympathetic activity. The sympathetic response, however, does not lead to a uniform change in all sympathetically innervated systems. The response magnitude of specific systems, such as the cardiovascular system, depends to a large extent on constitutional and hereditary factors. The subjective awareness of bodily changes accompanying heightened sympathetic activity is inaccurate; people often recognize the direction but not the degree of change. The level of body awareness depends on various psychologic factors, of which anxiety plays an important role. Acute stress produces sympathetic activation in nonanxious as well as anxious persons. Nonanxious persons tend to have more flexible autonomic responses. They show stronger responses to novel situations but return to lower autonomic levels earlier and habituate faster than do anxious persons. That is, nonanxious persons possess a greater autonomic flexibility than anxious persons. It is important to know the physiologic state of anxiety disorder patients during periods when they do not feel anxious, during times of heightened tension, during the performance of standardized stress tasks, during exposure to psychopathology-specific stressors, and during "spontaneously" occurring surges of anxiety, such as panic attacks. At present only limited information concerning these conditions is available. There is little evidence that anxiety disorder patients, perhaps with the exception of very severe cases, have an increased sympathetic tone when they do not feel anxious. However, all anxiety disorders, with the exception of simple phobia, show some sort of physiologic activation in threatening situations, including the recording of physiologic baseline values in laboratories. The type of activation differs among anxiety disorders. During periods of rest, social phobics and panic disorder patients tend to show sympathetic activation, generalized anxiety disorder patients show increased muscular tension without sympathetic activation, and obsessive-compulsive patients show increased muscular tension along with sympathetic inhibition. Under laboratory stress, both normals and anxiety disorder patients react with sympathetic arousal. However, in generalized anxiety and obsessive-compulsive patients the response is weaker than in normals, suggesting the presence of an inhibitory process. Thus, the autonomic flexibility of anxiety disorder patients is reduced.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3047708 TI - Anxiety and gastrointestinal illness. AB - The substrate for modulation of the gastrointestinal tract by anxiety and other psychiatric disorders is provided by our knowledge of central nervous system control mechanisms of gastrointestinal functions. Recent experiments that examine the gastrointestinal response to acute stress amplify prior work and confirm that central stimuli, viz. emotional stress, can produce measurable gastrointestinal changes. These changes are not uniformly predictable on the basis of knowledge of the control mechanisms, alone. Acute stress experiments cannot be directly extrapolated to anxiety disorders and their relationship to gastrointestinal illness; careful physiologic studies in these more chronic disorders are sparse. Anxiety disorders and other psychiatric illnesses are particularly common in patients with the functional bowel diseases. This heterogeneous group of gastrointestinal diseases remains fertile territory for examining emotion-gut relationship on a biologic level. PMID- 3047707 TI - Panic disorder, cardiology patients, and atypical chest pain. AB - Although patients with angiographically normal coronary arteries have low mortality, several studies have indicated that their social and work morbidity is high. Panic disorder appears to be a major contributor to the continuing chest pain in this population. There are also many chest pain patients appearing in cardiology clinics who also do not have heart disease but who are not given the opportunity to be evaluated for psychiatric disorders. Among those presenting with atypical or nonanginal chest pain, panic disorder represents a likely etiologic consideration. The fact that such patients do exist in cardiology populations is further substantiated by an open-label trial of alprazolam which demonstrated a positive effect in patients selected from those with atypical chest pain and no heart disease found to fit panic disorder criteria. These findings strongly support the increasing affiliation between cardiology and psychiatry and reinforce the belief that many problems of the heart may be problems of the mind/brain. PMID- 3047709 TI - Anxiety disorders in adults with diabetes mellitus. AB - Anxiety disorders are common among patients with diabetes and appear to recur in a substantial proportion of cases. The presence of generalized anxiety disorder is associated with both poorer glucose control and the increased report of clinical symptoms of diabetes. A complex interactive model best accommodates observations from psychiatric-diabetic research, one wherein diabetes may affect psychiatric status or vice versa in an independent or reciprocal fashion. No information from systematic studies is available currently to determine if treatment of the psychiatric disorder will have a beneficial impact on glucose regulation, despite the direct clinical relevance of such information to management of diabetes. PMID- 3047710 TI - Panic disorder and mortality. AB - Evidence so far indicates two sources for excess mortality in panic disorder- suicide and cardiovascular morbidity. The risk for eventual suicide may rival that for primary depression, but the predictors and the necessary antecedents probably differ. The lapse between diagnosis and suicide may be larger for panic disorder, and complications such as secondary depression and substance abuse may be necessary. There are few well-established predictors for primary depression despite many relevant studies. The risk for suicide in panic disorder is barely recognized, and established predictors are accordingly remote. One study has demonstrated excess cardiovascular mortality among males with panic disorder, and another from the same center has provided weak support. Only one additional study has provided the necessary detail as to sex and cause, and those findings were quite supportive, although the subjects may have been mixed diagnostically. There are numerous feasible explanations for excess cardiovascular mortality in panic disorder and even some reason to believe that successful treatment might lessen it. To so advise patients would be not only premature at this point but unnecessary and countertherapeutic--unnecessary because these patients are motivated by discomfort to seek treatment and countertherapeutic because cardiovascular morbidity is what many of these patients pathologically fear. Rather, the findings suggest focus for future study. The initial findings of excess cardiovascular morbidity in males badly need replication, as do the more recent findings of Kahn et al. Likewise, animal models may reveal some of the pathophysiologic mechanisms at work. It is hoped that these efforts will converge in the not-too-distant future. PMID- 3047711 TI - Panic disorder and major depression. A tale of two syndromes. AB - Considerable research has been conducted to clarify relationships between panic disorder and major depression. From a number of perspectives, it now appears that panic disorder and major depression are not identical illnesses. While many patients with panic disorder are likely to experience an episode of major depression at some point during their lives, the timing of this occurrence is highly variable. While depression may be an expected and understandable result of having to live with chronic anxiety and phobic avoidance, the available evidence suggests that such a hypothesis is not particularly tenable. Depression can occur in individuals with or without severe agoraphobia and in individuals ill with panic for greater or lesser periods of time. Comorbidity in panic, particularly for social phobic or obsessive-compulsive symptomatology, does serve as a risk factor for the lifetime occurrence of depression and may denote a more severe illness. Biologic markers (Table 4), while of limited diagnostic utility in clinical practice, may reveal important pathophysiologic similarities and differences between panic disorder and major depression. Current evidence points to many areas of biologic overlap, with some important areas of independence (Fig. 7A). Although not extensively discussed in this chapter, several clinical parameters paint an analogous picture (Fig. 7B). There is a need for future studies of biologic markers in individuals over time, in various phases of illness. Furthermore, the study of multiple biologic markers in the same individuals would be a worthwhile pursuit, perhaps leading toward the delineation of underlying pathophysiologic mechanisms. In summary, then, we favor a conceptualization of panic disorder and major depression as nonidentical disorders with many shared characteristics. Future studies as suggested above, particularly when coupled with the power of genetic studies not described in this chapter, may eventually lead to a clearer demarcation of the boundaries between these two intriguing psychiatric syndromes. PMID- 3047712 TI - Biologic systems and their relationship to anxiety. Summary. AB - Substantial progress has been made in understanding the relationships between clinical anxiety and biologic systems. Optimism must be tempered, however, by the realization that lofty speculations more often than not wither under scientific scrutiny. PMID- 3047713 TI - [Neurological problems in children with regard to so-called risk factors during pregnancy and labor]. PMID- 3047714 TI - Immunohistochemical and electron microscopic techniques in the diagnosis of viral encephalitides. AB - Electron microscopy and in particular, immunohistochemical examination techniques have substantially improved the chances to attain an etiologic diagnosis in viral encephalitis. The application of these techniques is needed first of all in the examination of cerebral biopsy specimens, but they can contribute to the clearing of the etiology at post mortem study of the brain, too. This review outlines the basic techniques and their application fields. In immunohistochemistry beside the demonstration of specific viral antigens the analysis of the humoral and cellular immune reactions and the identification of the cell types involved by cell specific markers is also important. The second part of the review gives a synopsis of the results of electron microscopic and immunohistochemical studies in the most important acute, subacute and chronic encephalitides occurring in Europe. PMID- 3047716 TI - Rudolf Virchow and modern aspects of tumor pathology. AB - Rudolf Virchow (1821-1902), the great German pathologist of the 19th century, founder of the "Zellularpathologie", also dealt with principal problems of tumor pathology. His theoretical concepts concern the definition and characterization of the tumor process, the dignity and diagnosis, published especially in his 3 monographs "Die Entwicklungsgeschichte des Krebses" (1847), "Handbuch der speciellen Pathologie und Therapie" (1854) and "Die krankhaften Geschwulste" (1863-65). The achievements of Rudolf Virchow are not related to the description and diagnosis of specific tumors together with their genesis and interpretation. There have been quite a few errors and false concepts which can longer be accepted today. The timeless modernity and topicality of Rudolf Virchow's postulations on tumor pathology is based on his general conclusions which as key propositions have retained general validity throughout more than a century. They have been confirmed and further interpreted by knowledge which has been accumulated largely over the past one or two decades in the fields of tumor research, electron microscopy, molecular biology, genetics, and immunology. PMID- 3047715 TI - An immunohistochemical study of normal human neonate and adult parotid gland tissue. Detection of lysozyme, lactoferrin, a1-antichymotrypsin, a1-antitrypsin and carcinoembryonic antigen. AB - The immunohistochemical detection and distribution of lysozyme (Ly), Lactoferrin (Lf), a1-Antichymotrypsin (a1-AChy), a1-Antitrypsin (a1-AT) and Carcinoembryonic antigen (CEA) were studied in neonate and adult parotid gland tissue, using the peroxidase-antiperoxidase (PAP) method. Ly stain in neonates extended into acini, intercalated ducts and occasional cells of large ducts, whereas in adults Ly was usually confined to the intercalated ducts. The distribution pattern of Lf in neonates varied considerably between individual glands showing three staining patterns. Most of the intercalated ducts, some groups of acini and rare striated duct cells were positive for Lf in adults. a1-AChy and a1-AT in neonates were positive mainly in the large ducts, whereas staining for a1-AChy and a1-AT in adults frequently extended into some intercalated duct cells, although less intensively. Finally, CEA in neonates was localized in the lumina and luminal membranes of the acini, in intercalated ducts, and less frequently in the large ducts. In adults CEA was present predominantly in the lumina and luminal membranes of the intercalated duct cells. These differences may suggest an immunohistochemical postnatal differentiation of the parotid gland. PMID- 3047717 TI - Estramustine inhibits monocyte phagocytosis. AB - Estramustine phosphate (EMP) influence on human monocyte phagocytosis of fluorescein isothiocyanate (FITC)-labeled yeast cells was measured in vitro. The method used, a modification of Hed's technique (FEMS Microbiol Lett 1:357, 1977), can differentiate between yeast cell engulfment and adherence to the phagocytotic cell surface. EMP is now accepted in the treatment of advanced prostatic carcinoma. In concentrations corresponding to the clinical situation (20-40 micrograms/ml), it dramatically inhibited the process of phagocytosis. The engulfment phase was inhibited, whereas cell adherence was less affected. This might be due to direct interaction with the microtubule system. The effects were totally reversible. In contrast, the metabolites estradiol and normustine did not affect engulfment of yeast cells, either as single agents or combined. The results demonstrate that the EMP complex caused an impaired phagocytosis, which could be of pathophysiological significance in the compromised cancer patients. PMID- 3047718 TI - Prophylaxis of deep venous thromboembolism. Literature review. AB - The prophylaxis of deep venous thrombosis is critically related to potentially life-threatening pulmonary embolism. Each of the available agents has benefits and drawbacks which are reviewed in this article. While the search for the ideal method of prophylaxis continues, the appropriate agent appears to be one that balances the risk of complication with the potential risk of development of thromboembolism. Recommendations for acceptable prophylaxis are made for various procedures. PMID- 3047719 TI - Hypoglycaemia following treatment of hyperkalaemia with insulin and dextrose. AB - We describe four patients who developed symptomatic hypoglycaemia following treatment of hyperkalaemia with insulin and dextrose. Two patients had a delayed onset of hypoglycaemia, between 5 and 6 hours after treatment despite use of a 'soluble' type of insulin. A review of the literature revealed a variety of insulin and dextrose regimes but no research to assess the metabolic effects of such therapy in patients with renal failure. The possibility of significant hypoglycaemia following use of insulin and dextrose in the recommended dosages is rarely mentioned. The cases we describe demonstrate that there is no 'correct' dose of insulin and dextrose to suit every circumstance. Regular blood glucose estimations should be performed in all patients receiving such therapy for hyperkalaemia. PMID- 3047720 TI - Cerebral scintigraphy--the phoenix rises again. AB - This paper reviews the development of cerebral scintigraphy from its early days of planar imaging with simple technetium-99m labelled compounds to the recent revival of the technique in the form of positron-emission and single-photon emission tomography. A short explanation of instrumentation and radiopharmaceuticals is given as a prelude to a description of both techniques in normal and pathological situations. Particular emphasis is placed on the more readily-available single-photon emission-tomographic techniques using labelled amines in the functional investigation of disorders not readily diagnosed by computed tomography. PMID- 3047721 TI - Disseminated mucormycosis due to Cunninghamella bertholletiae in a liver transplant recipient. AB - Disseminated mucormycosis occurred in a 19 year old female following orthotopic liver transplantation for fulminant Wilson's disease. The causative organism Cunninghamella bertholletiae has previously been described in ten clinical cases, but never before in this setting. PMID- 3047722 TI - Penile hyperexcitability with recurrent ejaculations as the presenting manifestation of a case of rabies. AB - A patient with rabies presented with penile hypersensitivity and recurrent ejaculation. This preceded the more classical manifestations such as hydrophobia and aerophobia by approximately 8 hours. A review of the literature reveals that a variety of urogenital symptoms may rarely occur in rabies. Ours is however the first report documenting the occurrence of these symptoms at the onset of the disease in an autopsy proven case of rabies. Damage to the amygdaloid nucleus has been postulated to be the basis of these urogenital symptoms. This correlates well with our autopsy findings of extensive neuronal damage and Negri bodies in the temporal poles. PMID- 3047723 TI - Aspirin-induced gastroduodenal injury and its prevention by prostaglandins. AB - It appears likely that aspirin injury and thus bleeding is due to both inhibition of the mucosal synthesis of protective endogenous prostanoids and to a local irritant effect. This injury can be abolished in humans by acid-inhibitory doses of prostaglandins and can probably be ameliorated by 'cytoprotective' doses. It has yet to be shown that the gastropathy seen in patients taking long-term aspirin-like drugs can be prevented or treated by prostaglandins. The clinical relevance of such protection has not yet been established. PMID- 3047724 TI - Endoscopy in the management of upper gastrointestinal disease. PMID- 3047725 TI - Long-term treatment of duodenal ulcer. AB - Eighty per cent, or more, of duodenal and gastric ulcers which have healed after a course of treatment with drugs relapse when therapy is discontinued. Only gastric operations or continuous maintenance treatment with H2 receptor antagonists have so far been shown to produce sustained remission in the majority of patients. Maintenance treatment with drugs is safer than gastric surgery, since the adverse effects of medical therapy are uncommon, mild and reversible while the after-effects of surgery are common and may be severe and irreversible. The maintenance treatment of ulcer patients must be continued for many years and may, perhaps, have to be life-long. PMID- 3047726 TI - Surgical treatment of duodenal ulcer. AB - This paper reviews the pathophysiology of peptic ulcer disease, the role of the vagus and gastrin in the control of gastric acid secretion and motility and the rationale for modern operations for peptic ulcer. The importance of arriving at a balanced decision before recommending surgery for a peptic ulcer is discussed. The indications for surgery in elective and emergency cases are defined and the results reviewed critically and compared with medical treatment. Novel operative procedures are mentioned but still await validation follow-up studies before their final assessment. Well tried operations should not be dismissed lightly and the importance of prospective randomized trials to evaluate new procedures is emphasized. PMID- 3047727 TI - Benefits and costs of lifestyle change to reduce risk of chronic disease. AB - Individuals do not benefit equally from attempts to change their lifestyles in an effort to lower their risk for disease or to improve their quality of life. A change in one lifestyle behavior may cause an increase in another risk factor and reduce the benefits of the anticipated change. The social environment exerts pressures and makes available resources that also influence the benefits and costs of a particular health behavior change. These pressures and resources vary depending on the individual and his or her social context. This article uses the target behavior of smoking as an example of a lifestyle change and considers the benefits and costs that interventionists need to be aware of if they are to effectively facilitate health behavior change. This approach requires the identification of resources at different levels of the environment (e.g., family, community, institutions) that may influence the cost/benefit ratio. Such an analysis is appropriate whether one is considering a model of individual behavior change or a public health model that seeks to intervene at the community-wide level to promote health and reduce disease risk among a large segment of the population. Specific recommendations based on this approach are offered and it is concluded that both individual and public health approaches are necessary to achieve optimal health behavior change in our population and to optimize the cost/benefit ratio of such change for all individuals. PMID- 3047728 TI - [Contradictions in the modern theories of endocrinology]. PMID- 3047730 TI - [Diagnosis and control of diabetes mellitus]. PMID- 3047729 TI - [The human insulin gene and diabetes mellitus]. PMID- 3047731 TI - [Complications in treatment with peroral antidiabetic preparations]. PMID- 3047733 TI - [Dynamic regulation of steroid hormones by steroid-modulating agents through their complexing with intracellular proteins]. PMID- 3047732 TI - [Prostanoids in diabetes mellitus in children]. PMID- 3047734 TI - [Significance of dihydrotestosterone for the development of the gonads and the reproductive function in the male]. PMID- 3047735 TI - [The importance of prolactin in the mechanism of male puberty]. PMID- 3047736 TI - [Diagnostic value of determining beta 2 microglobulin in patients with renal tuberculosis]. PMID- 3047737 TI - [New methods of the surgical treatment of tuberculous spondylitis in children]. PMID- 3047738 TI - [The 40th anniversary of the World Health Organization]. PMID- 3047739 TI - [Tuberculosis of farm animals and man]. PMID- 3047740 TI - Masters' programs in nursing--cardiovascular specialization. PMID- 3047741 TI - Arrhythmias and sleep pattern disturbances in cardiac patients. PMID- 3047742 TI - Combined procedure of distance geometry and restrained molecular dynamics techniques for protein structure determination from nuclear magnetic resonance data: application to the DNA binding domain of lac repressor from Escherichia coli. AB - The technique of two-dimensional nuclear magnetic resonance (2D-NMR) has recently assumed an active role in obtaining information on structures of polypeptides, small proteins, sugars, and DNA fragments in solution. In order to generate spatial structures from the atom-atom distance information obtained by the NMR method, different procedures have been developed. Here we introduce a combined procedure of distance geometry (DG) and molecular dynamics (MD) calculations for generating 3D structures that are consistent with the NMR data set and have reasonable internal energies. We report the application of the combined procedure on the lac repressor DNA binding domain (headpiece) using a set of 169 NOE and 17 "hydrogen bond" distance constraints. Eight of ten structures generated by the distance geometry algorithm were refined within 10 ps MD simulation time to structures with low internal energies that satisfied the distance constraints. Although the combination of DG and MD was designed to combine the good sampling properties of the DG algorithm with an efficient method of lowering the internal energy of the molecule, we found that the MD algorithm contributes significantly to the sampling as well. PMID- 3047743 TI - Structural comparison of the prokaryotic ribosomal proteins L7/L12 and L30. AB - The structures of two prokaryotic ribosomal proteins, the carboxyterminal half of L7/L12 from Escherichia coli (L12CTF) and L30 from Bacilus stearothermophilus display a remarkably similar fold in which alpha-helices pack onto one side of an antiparallel, three-stranded, beta-pleated sheet. A detailed comparison of the structures by least-squares methods reveals that more than two-thirds of the alpha carbons can be superimposed with a root mean square distance of 2.33 A. The principal difference is an extra alpha-helix in L12CTF. The sequences of the proteins display a distinct conservation in regions which are crucial to the common fold, in particular the hydrophobic core. It is proposed that the similarity is a result of divergent evolution. PMID- 3047744 TI - Dopaminergic modulation of some central actions of angiotensin II in vivo. AB - Studies were performed in 12 conscious sheep of both sexes to determine if a brain dopaminergic pathway is involved in modulating the central actions of angiotensin II (Ang II) in regulating body temperature and plasma renin activity (PRA). Previous data showed that intracerebroventricular (ICV) infusion of Ang II significantly decreased PRA and body temperature. In contrast, converting enzyme inhibitor SQ 20881 (SQ) or dopamine (DA) significantly increased PRA and body temperature of sheep. In the present study, ICV infusion of the DA antagonist metoclopramide (MCP) (20 micrograms/min) significantly decreased PRA to 68 +/- 5% of the basal level. When sheep were pretreated with ICV MCP (20 micrograms/min) for 2 hr and then infused ICV with MCP (20 micrograms/min) plus DA (20 micrograms/min), Ang II (25 ng/min), or SQ (1 microgram/min), the PRA and temperature responses to DA, Ang II, or SQ were all abolished or attenuated significantly. The converse did not hold. Sheep pretreated with SQ (1 microgram/min) still showed a significant increase in body temperature (0.43 +/- 0.05 degree C) when infused with DA (20 micrograms/min). These results support the hypothesis that a central DA pathway is involved in the modulation of the actions of centrally administered Ang II on temperature and PRA. PMID- 3047745 TI - Role of prostaglandins in controlling plasma fibronectin levels. AB - Fibronectin is a normal glycoprotein component of plasma, interstitial fluid, and extracellular matrix which has binding sites for collagen, gelatin, actin, glycosaminoglycans, fibrin, Staphylococcus aureus, and some cells. Since it is a dimer, it can crosslink these substances to each other or to extracellular components of basement membrane, thereby affecting many physiological processes. The level of circulating fibronectin is markedly reduced following even moderate blunt or operative trauma, thermal injury, starvation, advanced cancers, hemorrhage, etc. Replacement therapy has been tried with some success in patients who become septic following multiple injuries. The reduction in plasma fibronectin has been attributed to several causes including consumption by binding to cell debris at the site of injury, binding to circulating cell debris and its subsequent removal by elements of the phagocytic system, and degradation by proteolytic cleavage. However, the amount of fibronectin removed from circulation raises some question about this. In this paper, we used indomethacin, ibuprofen, imidazole, and essential fatty acid deprivation to inhibit the synthesis of prostaglandins in young adult rats. Thirty minutes after ip administration of one of the inhibitors, the rats were subjected to a midline laparotomy and mild intestinal manipulation. Blood samples were taken at intervals following closure of the incision and analyzed for fibronectin. In all cases, the normal decline in plasma fibronectin seen in untreated rats was abrogated by inhibiting prostaglandin synthesis. Since imidazole specifically inhibits thromboxane A synthesis, this strongly suggests that thromboxanes directly or indirectly control the trauma-induced reduction in circulating fibronectin. This was confirmed by ip injection of thromboxane into the rats which resulted in a decline in plasma fibronectin levels. PMID- 3047746 TI - Regulation of sodium pump abundance in animal cells. PMID- 3047747 TI - Is the constitutive Kfp-K+ pumping system of E. coli able to adapt to osmotic shifts without new protein synthesized? PMID- 3047748 TI - Regulation of muscle contraction and relaxation in heart. AB - The calcium uptake and release machinery in heart SR have been characterized: (1) The calcium pump membrane is involved in energized Ca2+ uptake enabling muscle to relax. The calcium pump protein (CPP) in heart SR is modulated by protein kinase phosphorylation of phospholamban lowering the KCa2+. We conclude that in the membrane, phospholamban elevates KCa2+ of calcium pump protein. Phosphorylation of phospholamban attenuates the influence of phospholamban. In the limit, the intrinsic KCa2+ of calcium pump protein in heart and skeletal muscle are approximately the same. (2) The junctional face membrane is involved in calcium release which triggers muscle contraction. Ryanodine is a specific modulator of the Ca2+ release channels of SR which are involved in excitation-contraction coupling. The ryanodine receptor has been isolated, found to be equivalent to the feet structures, and on reconstitution into bilayers, identified as the calcium release channel of SR. The calcium release channel of SR is closed by ruthenium red and Mg2+ and opened by Ca2+ and ATP and low ryanodine concentration. The calcium release channel of SR is not effected by drugs such as nitrendipine, diltiazem and D-600 which modulate the slow inward Ca2+ channel of the plasmalemma/transverse tubule. (3) The calcium release channels from heart and skeletal muscle SR are similar but not identical (Table IV). Important differences distinguish the calcium release machinery in heart from that of skeletal muscle. 1. In heart there are two sources of calcium fluxes: a) extracellular Ca2+ enters via the plasmalemma slow inward calcium current; and b) "Ca2+ induced Ca2+ release" from SR. In skeletal muscle, SR is the single main source of calcium which enters via "Depolarization induced calcium release". 2. The calcium release channel from heart SR has a lower Mr approximately 340,000 vs 360,000 for skeletal muscle. 3. Ryanodine binding in cardiac SR is distinct from that in skeletal muscle (Fig. 7). 4. The isolated calcium release channel from heart SR is more sensitive to Ca2+ for calcium release (Hymel et al. 1988c). Significant progress has been achieved in identifying the calcium release channel of SR in heart and skeletal muscle. The focus of excitation-contraction coupling now shifts to defining the precise nature of the coupling of excitation to contraction. PMID- 3047749 TI - Image analysis of the Escherichia coli lactose permease molecule in filamentous arrays. PMID- 3047751 TI - The role of prostaglandins in the regulation of cell proliferation. PMID- 3047750 TI - Eicosanoids in the CNS: sources and effects. PMID- 3047752 TI - The roles of essential fatty acids in the development of diabetic neuropathy and other complications of diabetes mellitus. PMID- 3047753 TI - Prostacyclin-enhanced calcium sequestration by microsomes isolated from rat left ventricle. AB - The objective of these experiments was to investigate the direct effect of prostacyclin on calcium binding and uptake by microsomal vesicles isolated from rat left ventricle. The protein content of the preparation was found to be 2.73 +/- 0.18 mg microsomal protein/g of left ventricular wet weight. Steady-state calcium binding and uptake by microsomal vesicles was reached at 6 minutes in control animals and 20 sec in prostacyclin-treated experiments. Prostacyclin increased steady-state calcium binding and uptake from a control of 52.48 +/- 4.16 up to 109.31 +/- 3.06 nmol/mg protein (p less than 0.05) and from 238.07 +/- 12.37 up to 314.85 +/- 1.23 nmol/mg protein (p less than 0.05) respectively. Doubling the concentration of prostacyclin from initial dose of 1.2 x 10(-8) M did not alter calcium binding and uptake further. PMID- 3047754 TI - Some cautionary comments on the use of retrospective designs to evaluate treatment efficacy. AB - This article contrasts the true experimental approach with nonexperimental alternatives, such as retrospective designs, for research on treatment efficacy. The potential problems and limitations of the nonexperimental approach are highlighted so that when deviations from the more desirable experimental methodology are necessary, they can be undertaken with full knowledge of the hazards to be expected. Important issues are illustrated through discussion of a specific type of retrospective design, the case-control design. PMID- 3047755 TI - Needs of the elderly and the politics of health care. The social context of changes in the delivery system. AB - The emergence of the elderly as a substantial subgroup within the population has been identified as signaling a crisis for the health care system. This article places recent changes in health care financing for the elderly in the context of biomedical, demographic, and social factors. These factors, in turn, are related to the larger economic and political structures that have shaped our national health care policies and programs. Current policies and programs are inadequate in meeting the needs of the elderly because they provide a limited array of services. This article also examines how the needs of the elderly have been portrayed to support age-based entitlements to limited health care coverage, irrespective of need across age strata. Physical therapists can use their understanding of the genesis of particular public policies to assist in developing a health care system that is responsive to the needs of all members of society. PMID- 3047756 TI - Decision analysis using a spreadsheet. PMID- 3047757 TI - The history of augmentation mammaplasty. PMID- 3047759 TI - Vectors for expression of truncated coding sequences in Escherichia coli. AB - We describe the construction of vectors for expressing in Escherichia coli DNA fragments obtained by progressive deletions of DNA inserts in single-stranded sequencing vectors as M13 or pTZ according to the methode of Dale et al. (Plasmid 1985, 13, 31-40). These vectors, pIMS1, pIMS5, and pIMS6, harbor all of the elements required for the regulated expression of any open reading frame flanked by EcoRI restriction sites. The encoded peptides contain only a few vector derived amino acids. A method is described for direct selection of recombinant clones by in situ RNA hybridization. The properties of the expression vector have been analyzed with a DNA deletion series obtained from the cDNA coding for the regulatory subunit of Dictyostelium discoideum cAMP-dependent protein kinase. PMID- 3047758 TI - A single amino acid difference between Rep proteins of P1 and P7 affects plasmid copy number. AB - P1 and P7 are closely related plasmid prophages which are members of the same incompatibility group. We report the complete DNA sequence of the replication region of P7 and compare it to that of P1. The sequence predicts a single amino acid difference between the RepA proteins of these two plasmids, no differences in methylation sites or regions where dnaA protein is expected to bind, and no difference in the spacing of the major features of the two replicons. A P1 replicon with a mutation in repA, the gene that encodes an essential replication protein, is complemented for replication by providing either the P1 RepA protein (RepA1) or the P7 RepA protein (RepA7) in trans. Furthermore, when either of these proteins is supplied in trans, the plasmid copy number of P1 cop mutants drops to that of P1 cop+. However, when RepA7 is supplied, the copy number of P1 cop and P1 cop+ is higher than that when RepA1 is supplied. This indicates that the single amino acid difference between the two versions of the RepA protein plays an important role in determining the plasmid copy number. PMID- 3047760 TI - Electrical stimulation of onlay bone grafts. AB - An animal model was developed to determine the ability of capacitively coupled electrical fields to enhance onlay bone graft survival in the craniofacial skeleton. Fifteen male New Zealand white rabbits were divided into control and stimulated groups. Blocks of iliac bone were transplanted as onlay grafts to the mandibular rami. In all animals a capacitor apparatus was attached externally over the right mandibular ramus; however, a 5-V peak-to-peak sinusoidal signal was applied only in the stimulated group. The experimental period was 6 weeks, with a total of 30 days of constant stimulation. Graft resorption in the stimulated animals was decreased a total of 24.8 percent (p less than 0.001). There appeared to be a trend toward decreased graft resorption caused by the apparatus alone, although this was not statistically significant. The electric field alone decreased resorption by 11.5 percent (p less than 0.05). No distinguishing features were demonstrated by fluorescent vital stains or routine histology. PMID- 3047761 TI - Skin grafts for the burned palm in children. PMID- 3047762 TI - Factors affecting psychiatrists' availability to serve in public programs. AB - This paper examines the factors contributing to reduced psychiatric participation in community mental health centers and state hospitals. Important considerations include level of compensation, other incentives, experiences during medical education, affiliation with medical settings, and licensing requirements. Suggestions are offered which may improve recruitment and retention of psychiatrists, as well as increase psychiatric involvement in the public mental health sector. PMID- 3047763 TI - Linked changes in mental, health service delivery and psychiatric education. AB - In sociocultural perspective, this paper traces the impact of deinstitutionalization and the community mental health movement on the mental health service delivery system and subsequent effects on psychiatric training. Examples are given of changes at the national, state, and local levels. The current need is for new initiatives to interest residents in public sector psychiatry and in work with the chronic patient. Issues discussed include new conceptions of the parameters and roles of psychiatric treatment, involvement of families and other support groups in patient care, comprehension and applications of cultural values, and new functions of the state hospital in the continuum of care as locus of specialized services and of professional training and research. PMID- 3047764 TI - Moving treatment into the community: implications for psychiatry. PMID- 3047765 TI - The role of psychiatry in the public mental health delivery system: an introduction. PMID- 3047767 TI - Leo Rangell. An appreciation. PMID- 3047766 TI - Role of psychiatrists in the public mental health system. AB - This paper examines the contributions of the psychiatric profession within the public mental health system. Following a discussion of the current status of psychiatry and the public mental health delivery system, the author outlines a number of key recommendations regarding the appropriate role of psychiatrists in public systems. The author emphasizes the importance of the public mental health system in all areas of psychiatric practice. PMID- 3047768 TI - Dickens, Little Dorrit, and soul murder. AB - The effects of moderate psychological child abuse by parents and parental substitutes on Charles Dicken's life and work are examined. PMID- 3047769 TI - [Changes in the nuclear proteinase activity of the liver in gamma-irradiated rats]. AB - Activity of nuclear proteinases in rat liver was studied, by their capacity of splitting a casein substrate, 5-48 h following gamma irradiation. The proteinase activity of nuclei was shown to increase by more than two times in 2-5 h and to decrease by 3-4 times after 15-48 h compared to that of the nonirradiated controls. A sharp decrease in the proteinase activity was also noted in the nuclear matrix of irradiated rat liver. PMID- 3047770 TI - [Regulation of the activity of Ca2+-phospholipid-dependent protein kinases of the rat brain in acute radiation lesions]. AB - Regulation of activity of Ca2+-phospholipid-dependent protein kinases of rat brain cortex by calcium ions and phosphatidylserine was studied 60 min after whole-body X-irradiation (0.21 C/kg). Irradiation was shown to cause multiple disorders in regulation of activity of the enzymes under study. PMID- 3047771 TI - [Specificity of the lethal and mutagenic actions of pico-second laser pulses of 532-nm wavelength]. AB - A study was made of the lethal effect of pulse laser (second harmonic Nd+3:YAG laser of 532 nm, pulse length 3.3.10(-11) s, peak intensity from 4.10(12) to 1.10(14) W/m2) on HeLa cells at the phases of active and stationary growth, and lethal and mutagenic effects of this radiation on E. coli cells. As was shown, HeLa cells at both growth phases and E. coli cells exhibited low sensitivity to laser radiation at lambda = 532 nm. PMID- 3047772 TI - [Intensification of the excretory flow of plutonium-239 from the body in the late stage of its metabolism]. AB - The paper deals with the effect of iron preparations on the excretion of plutonium 239 from a body at a later stage of the radionuclide metabolism. The experimental results show that oral administration of the iron preparation at a later stage of 239Pu metabolism enhances the radionuclide excretion both in urine and in faeces. On the basis of the results obtained the coefficients are calculated for 239Pu excretion in urine and faeces and for its content in the organs of deposition. This may be used for increasing the sensitivity of indirect dosimetry of plutonium-239 within the body. PMID- 3047773 TI - [Biological action of plutonium-239 on Salmonella typhimurium]. AB - Salmonella typhimurium cells were exposed in a 239Pu citrate solution. Cell death and induction of gene mutations were an exponential function of gamma radiation dose. LD37 was 34.8 Gy; mutation doubling dose, 19 Gy. PMID- 3047774 TI - The injured spinal cord. Evaluation with magnetic resonance and intraoperative sonography. AB - MR is now the study of choice in evaluating both the acutely and previously injured spinal cord. A number of crucial diagnoses can be made with MR that are difficult to obtain otherwise. The discovery of acute disk herniation in association with a fracture/dislocation may indicate the need to remove the disk and stabilize the patient on a relatively urgent basis, and the approach to the stabilization procedure (either anterior or posterior) can be determined by the MR findings. Hemorrhage outside the cord may require immediate surgical attention, and identification of abnormal signals from the cord itself consistent with hemorrhage or edema may help to explain the patient's clinical status. Our experience indicates that obtaining an MR in acutely injured patients who have incomplete injuries is particularly helpful in their surgical management. The presence of intramedullary or extramedullary cysts in the previously injured spine can explain a worsening clinical picture and direct the surgeon to the proper area (or areas) for decompression. Flow-sensitive studies currently under evaluation are giving a greater insight into the dynamics of these cysts. Late decompression of cord tissue and roots requires accurate preoperative MR evaluation of possible bone or disk impingement on neural tissue. In surgery of both the acute and chronically injured spine, intraoperative sonography plays a crucial role. The adequate shunting of intramedullary and subarachnoid cysts, the confirmation of the efficaciously placed Harrington rods, the demonstration of removal of compressive bone or disk fragments, and the uncovering of important but unanticipated abnormalities such as subligamentous hematoma or an incidentally herniated disk all attest to sonography's value in the operating room. MR preoperatively and sonography intraoperatively are important tools utilized in the care of spine-injured patients. PMID- 3047775 TI - MR contrast agents: an overview. AB - In this article an overview of current and potential MR contrast agents is given. The mechanism of action of contrast agents and their relationship to both T1 and T2 relaxation are explored. Both paramagnetic and superparamagnetic substances are considered. Various physical states of these materials, including small ionic, lipophilic, and macromolecular forms are explored as possible contrast agents. Several clinical examples are given and speculation is made about the future potential of MR contrast agents. PMID- 3047776 TI - Nuclear medicine: SPECT comparisons to PET. AB - Early comparisons between PET and SPECT imaging of the brain, therefore, confirm that SPECT is capable of revealing information about regional brain function and metabolism at a much lower cost and degree of complexity. While PET will continue to play a leading role in research, SPECT imaging will increasingly be used clinically to evaluate functional disorders of the brain. PMID- 3047777 TI - Positron emission tomography in the investigation of central nervous system disorders. AB - Positron emission tomography is a noninvasive tomographic technique for measuring regional tissue concentrations of labeled radionuclides in man. Detection of two photons emitted from the annihilation of a positron and an electron is used to reconstruct the distribution of a positron-emitting isotope within an organ. PET provides the capacity to measure quantitatively the local tissue distribution of a variety of radionuclides that are attached to compounds that distribute according to function. Commonly measured functions include local cerebral metabolism using 18F-fluorodeoxyglucose or 11C-deoxyglucose, cerebral blood flow, cerebral oxygen utilization, and cerebral blood volume. Clinical applications of PET are multiple, involving normal and disease states. By demonstrating the metabolic alterations, PET adds another dimension to our understanding of the brain, which up until recently has been based on the structural changes seen on CT and MRI. PMID- 3047778 TI - Infant cranial sonography. AB - Sonography is well recognized as an excellent method of imaging the infant brain. A detailed discussion of the sonography of the preterm patient forms the first portion of this article. The numerous other clinical situations in which sonography can be used either alone or in conjunction with CT follow. PMID- 3047779 TI - Cerebrovascular ultrasound imaging. AB - Advancements in technology have brought noninvasive diagnosis of cerebrovascular disease to a high level of accuracy in a relatively short period of time. Conventional duplex sonography allows precise vessel localization and Doppler quantification. The further refinement of color flow imaging promises to simplify the examination and add considerable confidence to the diagnosis. Regardless of the technology used, ultrasound evaluation of the carotid arteries remains a difficult examination to perform, and excellent results are only possible with meticulous technique. The astute sonographer or sonologist will use all aspects of available ultrasound technology to the fullest. Careful evaluation of a seemingly overwhelming array of parameters will result in an impressive degree of diagnostic accuracy. PMID- 3047780 TI - Surgical neuroangiography of the spine and spinal cord. AB - The SCAVMs are high-flow lesions that present at a younger age, during the second or third decades of life. The most common presenting symptoms are subarachnoid hemorrhage and hematomyelia. Endovascular treatment of these lesions has become an important adjunct to surgical management. Complete occlusion is possible by superselective catheterizations and injection of a liquid embolic material. The SDAVFs are slow-flow AV shunting at the dura, which causes progressive neurologic deficit in older age--the fifth or sixth decades of life. Embolization is now the primary mode of treatment. Preoperative embolization of the hypervascular tumors of the spine and spinal cord has become a necessity to reduce bleeding during surgery, and it even reduces the size of the tumor and relieves spinal block. Thorough knowledge of vascular anatomy and better understanding of hemodynamics of these lesions are essential to perform proper and safe embolization. PMID- 3047781 TI - Image-guided pleural biopsies: indications, technique, and results in 23 patients. AB - Twenty-six pleural biopsies were performed on 23 patients over a 3-year period. Twenty-three biopsies were performed guided with ultrasound; one, with computed tomography; and two, with fluoroscopy. Indications for an image-guided pleural biopsy were (a) pleural masses or thickening that were either not seen on chest radiographs or seen only on one view and (b) small or loculated pleural effusions of unknown cause with no mass seen. If only pleural fluid was present, reverse bevel needles were used for biopsy (n = 15). If a discrete pleural mass or thickening was seen with cross-sectional imaging, standard (16-20 gauge) biopsy needles were used (n = 11). In the 23 patients, biopsy results were true positive in ten (nine with malignancy, one with tuberculous pleurisy), true negative in ten (confirmed either at subsequent thoracotomy or clinical follow-up), and false negative in three. Complications were few, with a significant pneumothorax occurring in two patients (8.7%). Image-guided biopsy of small pleural lesions and small pleural effusions can be performed by the radiologist who understands the special needles and techniques involved. PMID- 3047782 TI - Prenatal diagnosis of sacrococcygeal teratoma: sonographic-pathologic correlation. AB - The prenatal sonograms of 15 fetuses with sacrococcygeal teratoma were reviewed to determine the sonographic appearance and the role of sonography in the obstetric management. Each tumor appeared as a large mass arising from the fetal rump. The teratomas exhibited three sonographic patterns: nine were mixtures of cystic and solid components in equal proportions, four were predominantly solid with a few scattered anechoic areas, and two were unilocular cystic masses. Calcifications were detected in six cases. There was no correlation between the sonographic appearance and the presence of immature or malignant components. Ultrasonography allowed visualization of an intraabdominal component in six cases and assessment of findings that were of prognostic importance. Prenatal detection and size determination of the external component can play an important role in planning obstetric management because fetuses with a large tumor should be delivered by cesarean section to avoid dystocia and catastrophic hemorrhage during delivery. PMID- 3047783 TI - Suspected ectopic pregnancy: endovaginal and transvesical US. AB - Until the advent of endovaginal ultrasonography (US), transvesical US was the only US technique availab le for evaluation of patients with suspected ectopic gestation. A study was undertaken to assess the predictive ability of transvesical and endovaginal US and determine whether endovaginal US could be used alone. Fifty-three patients who had a positive pregnancy test finding and who were at risk for ectopic pregnancy were examined with both endovaginal and transvesical US. Twenty-nine were examined retrospectively and 24 were examined prospectively. Standard sonographic criteria were used to differentiate between intrauterine pregnancy and ectopic gestation. The clinical or pathologic diagnosis was ectopic pregnancy in 18 patients (34%), normal intrauterine pregnancy in 19 (36%), and abnormal intrauterine pregnancy in 16 (30%). Endovaginal US increased the sensitivity of detecting a live ectopic pregnancy (from 6% to 17%). Endovaginal US, by allowing early diagnosis of intrauterine pregnancy, significantly increased the diagnostic accuracy for ectopic pregnancy (from 60% to 83%). Endovaginal US provided significant additional information in women referred for sonography with a suspected ectopic gestation. On the basis of these findings it is concluded that endovaginal US can be used alone in the majority of women with suspected ectopic gestation. PMID- 3047784 TI - Percutaneous dilation of ureteral strictures in renal transplant patients. AB - Percutaneous dilation of benign ureteral strictures was performed as an alternative to surgical therapy in 14 patients with renal transplants. Dilations were performed with balloon catheters in 13 patients and with a tapered angiographic catheter in one patient. Eleven strictures were successfully dilated (79%). There were three recurrences (21%). Follow-up in nine of the 11 successful cases ranged from 12 to 61 months (mean, 29 months; median, 24 months). There were no complications directly related to balloon dilation. The high success rate in this series may be related to the early diagnosis of strictures in these closely followed patients. PMID- 3047785 TI - Peritoneal inclusion cysts: ovarian fluid in peritoneal adhesions. AB - In four women pelvic peritoneal inclusion cysts were diagnosed with ultrasound or computed tomography. Three patients had a history of multiple surgical procedures, whereas the fourth had prior severe abdominal trauma. Imaging studies showed large cystic structures contiguous with the adnexa. The normal ovarian appearance was distorted in two women. Pathologic confirmation was obtained in all cases. When large adnexal cystic structures are identified in young women with a history of surgery or trauma, the diagnosis of peritoneal inclusion cysts should be entertained. Recognition should result in conservative therapy rather than salpingo-oophorectomy. PMID- 3047786 TI - Digital image management networks: current status. PMID- 3047787 TI - Breast immobilization for occult mass aspiration. AB - Percutaneous needle aspiration of occult, presumably fluid-filled masses with ultrasound guidance is more difficult when the abnormality is in a deep location or when the breasts are pendulous or flaccid. The fenestrated compression plate available on dedicated mammographic units provides easy access for needle insertion and immobilizes the breast, permitting accurate puncture and aspiration. PMID- 3047788 TI - Color Doppler analysis of penile arteries in impotence. PMID- 3047789 TI - Thoracic empyema: management with image-guided catheter drainage. AB - Forty-three patients with thoracic empyema were treated by means of image-guided catheter drainage. In 40 patients, image-guided catheter drainage was the primary treatment method; in three it was used after conventional, surgical chest tube placement failed. Drainage was performed with ultrasound guidance in 30 patients (69.8%), computed tomography in eight (18.6%), and fluoroscopy in five (11.6%). A combination of guidance modalities was used in six patients. Image-guided catheter drainage alone was successful in 31 of 43 patients (72.1%). In three patients (7%), empyemas were initially drained, but a thoracotomy was ultimately required because of a persistent pleural peel. In eight patients (18.6%), the procedure failed, predominantly due to tube clogging, persistent pneumothorax, or progressive development of a pleural peel. In one patient, drainage was successful but he died 10 days later of complications of renal failure. No major complications were encountered. Treatment of these patients requires a thorough understanding of the pathogenesis of pleural space infection, principles of empyema drainage, techniques of abscess drainage under image guidance, and the use of a pleural drainage system. PMID- 3047790 TI - Pseudoaneurysms complicating organ transplantation: roles of CT, duplex sonography, and angiography. AB - In a retrospective study of proved pseudoaneurysms (PAs) in 15 patients with transplanted organs (11 liver, three kidney, one pancreas), the results of computed tomography (CT), duplex sonography, and angiography were reviewed. Of the 15 cases of PA, eight occurred at the arterial anastomosis and seven were nonanastomotic. Three of the eight anastomotic PAs were caused by infection. Of the seven nonanastomotic PAs, four were caused by percutaneous biopsy, two were caused by infection, and one was of undetermined cause. In nine (60%) of the 15 patients the PAs were incidentally detected at imaging studies performed for other reasons. Diagnosis requires a high degree of suspicion. CT was performed in nine cases and duplex sonography in ten. The diagnosis of PA was made with CT in six (67%) patients and with duplex sonography in five (50%). CT and duplex sonography could not enable diagnosis when the PA was small, when the arterial anastomosis was not included in the field of study, or when enhancement with intravenously administered contract material was suboptimal. Angiography depicted the PAs in all 15 patients. In three liver transplant recipients with gastrointestinal tract bleeding, the causative PAs were detected only with angiography. PMID- 3047791 TI - Stress, glucocorticoids and development. PMID- 3047792 TI - Actions of sex hormones on the brain: 'organization' and 'activation' in relation to functional teratology. PMID- 3047793 TI - Prenatal adverse effects of nicotine on the developing brain. PMID- 3047794 TI - Defects of neuronal migration and the pathogenesis of cortical malformations. PMID- 3047795 TI - Anticonvulsants and brain development. PMID- 3047796 TI - Neuropeptides and functional neuroteratology. PMID- 3047797 TI - Perinatal exposure to methadone: how do early biochemical alterations cause neurofunctional disturbances? PMID- 3047798 TI - Excitotoxic food additives: functional teratological aspects. PMID- 3047799 TI - Organometals and brain development. PMID- 3047800 TI - Concept of functional neuroteratology and the importance of neurochemistry. PMID- 3047801 TI - Prenatal stress effects on functional development of the offspring. PMID- 3047802 TI - Structural--functional relationships in experimentally induced brain damage. PMID- 3047803 TI - Coordination of cell development by the ornithine decarboxylase (ODC)/polyamine pathway as an underlying mechanism in developmental neurotoxic events. PMID- 3047804 TI - Brain cell acquisition and neurotropic drugs with special reference to functional teratogenesis. PMID- 3047805 TI - Implications of behavioral teratology for assessing the risks posed by environmental and therapeutic chemicals. PMID- 3047806 TI - Metabolic imbalance and nerve cell damage in the brain. PMID- 3047807 TI - Humoral and surface-anchored factors in development and repair of the nervous system. PMID- 3047808 TI - Gangliosides as cell adhesion factors in the formation of selective connections within the nervous system. PMID- 3047809 TI - Problems in studying functional teratogenicity in man. PMID- 3047810 TI - Transgenerational effects of drug and hormonal treatments in mammals: a review of observations and ideas. PMID- 3047811 TI - Behavioural teratology of exogenous substances: regulation aspects. PMID- 3047812 TI - The role of the insulin-like growth factors in the regulation of brain development. PMID- 3047813 TI - The long QT syndromes: a critical review, new clinical observations and a unifying hypothesis. PMID- 3047814 TI - Influence of ethanol on neuronal and synaptic maturation in the central nervous system--morphological investigations. PMID- 3047815 TI - Development of ventral spinal motoneurone fibres: a correlative study of the growth and maturation of central and peripheral segments of large and small fibre classes. PMID- 3047816 TI - [A love affair with phi X174. 2]. PMID- 3047817 TI - A clinical review of developments in the diagnosis and treatment of anorexia nervosa and bulimia. PMID- 3047818 TI - Clinical issues in the assessment of dementia and depression in the elderly. PMID- 3047819 TI - Psychiatric characteristics of patients undergoing cardiac transplantation. PMID- 3047820 TI - [Acute myelofibrosis, an open problem; immunohistochemical study of 2 cases]. PMID- 3047821 TI - [Usefulness of a bone marrow immunohistochemical study in multiple myeloma]. PMID- 3047822 TI - [Inhibition of the antihypertensive activity of intravenous captopril by pretreatment with indomethacin]. PMID- 3047823 TI - [Ultrasonographic aspects and course of hepatic cystic lesions with special reference to hydatid cysts]. PMID- 3047824 TI - Occult foreign body simulating a choroidal melanoma with extrascleral extension. AB - A 62-year-old man was noted on routine examination to have a dark lesion in the peripheral fundus of the right eye and a corresponding dark scleral mass. The lesion was initially suspected to be a choroidal melanoma with extrascleral extension. The patient denied having ocular trauma. Orbital x-rays and ultrasonography, however, demonstrated the lesion to be an occult transcleral metallic foreign body. An occult foreign body should be considered in the differential diagnosis of a small choroidal or ciliary body melanoma with extrascleral extension. PMID- 3047825 TI - Nanophthalmic uveal effusion. AB - A nanophthalmic patient with hypermetropia, shortened anterior-posterior axial length, and thickened choroid presented with a choroidal detachment and nonrhegmatogenous retinal detachment (NRRD). He underwent partial thickness, 5 x 7 mm, sclerectomies and 1-2 mm central sclerostomies 9.5 mm posterior to the limbus, specifically avoiding vortex veins. Complete resolution of the retinal and choroidal detachments occurred in spite of postoperative ultrasonograms demonstrating residual choroidal-scleral thickening. The effectiveness of this technique in our patient, in light of recent studies demonstrating histochemical and microscopic abnormalities in nanophthalmic sclera, suggests impairment of trans-scleral protein transport as the primary pathophysiologic mechanism in nanophthalmic uveal effusion. PMID- 3047826 TI - [Developmental adrenal tuberculosis. Value of x-ray computed tomography]. AB - Two cases of Addison's disease secondary to active adrenal tuberculosis are reported. Computed tomography showed hypertrophy of the adrenal gland, which was bilateral in one case and unilateral in the other. Repeat computed tomography scans during antituberculous chemotherapy demonstrated a progressive change of the adrenals toward atrophy and calcification, while the adrenal function remained impaired. The authors recall that adrenal tuberculosis may be unilateral first, then bilateral, and that the gland is initially hypertrophic before hormonal deficiency appears; later on, adrenal atrophy and calcification develop. Computed tomography seems to be useful in the aetiological diagnosis of Addison's disease. Moreover, it helps in determining whether or not antituberculous therapy is indicated, which is not always easy to decide in the absence of "active" focus. The finding, with or without positive tuberculin skin tests, of an adrenal hypertrophy unexplainable by any other pathology (e.g. metastasis, histoplasmosis) should call for antituberculous treatment, especially since it sometimes results in recovery of adrenal function. PMID- 3047828 TI - [Gonococcal infections of the joints]. PMID- 3047827 TI - [Osteonecrosis and their treatment]. PMID- 3047829 TI - [Thyroiditis]. PMID- 3047830 TI - [A case of organic hypoglycemia caused by a factor of unknown origin: low plasma insulin and IGF levels]. PMID- 3047832 TI - [Recurrent hypersomnia]. AB - Recurring hypersomnias are described according to 3 etiological groups: 1) idiopathic - the Kleine Levin syndrome and its clinical variants - 2) organic and 3) psychiatric. The typical form of the Kleine Levin syndrome is remarkable for the association of recurring episodes of sleep, overeating and temporary mental disturbances lasting from a few hours to several days. Its diagnosis is mainly based on clinical data Laboratory investigations have so far failed to document specific features. Emphasis is laid on circumstances at onset and pathological studies which could be in favour of a viral origin. Some clinical aspects and polysomnographic features are reminiscent of endogenous depression. The treatments of hypersomniac episodes based on stimulants are often disappointing. On the other hand, the prevention of the hypersomniac episodes of the Kleine Levin syndrome with lithium carbonate has been successful in several well documented cases as well as the prevention of the hypersomniac episodes of the menstruation related hypersomnia with ovulatory inhibitors. Organic and psychiatric forms of recurring hypersomnias are not well known. Their clinical features are described and their various possible etiologies indicated. PMID- 3047831 TI - [Chemotherapy in cancer of colon. General review]. AB - The results of surgery as sole treatment of colon cancer are summarized before dealing with those of chemotherapy. Curative or palliative chemotherapy remains controverted. The results of single drug treatments and of the conventional protocols with 5-fluorouracil and nitrosoureas have been disappointing. A promising approach is modulation of 5-fluorouracil by folinic acid, with a response rate of up to 45 p 100, and potentiation of 5-fluorouracil by cisplatin. Intra-arterial chemotherapy has been, and still is, an interesting method in liver metastases, but recent studies and the experience acquired with prolonged follow-ups have thrown doubts on some of its results, notably survival. Adjuvant chemotherapy is even more controversial than curative chemotherapy; however, a recent controlled study has yielded favourable results, and the best drug combinations have not yet been tested. It is concluded that cancers of the colon have shown some chemosensitivity, even though it has not reached the same level as that of cancers of other organs. PMID- 3047833 TI - [An adult case of Leigh's subacute necrotizing encephalomyelopathy]. AB - Leigh's encephalomyelopathy has been mainly observed in infancy and childhood. A later onset, during adolescence or adulthood has been rarely reported. Our patient was a 35 year-old man who died after 10 months of evolution of a subacute neurological syndrome, beginning with behavioural changes then a confusional state, epileptic fits, ataxia, autonomic disorders, abnormal alimentary behaviour and dementia. Diagnosis was only obtained by neuropathology, as in most of the published reports. However this diagnosis is suggested when exists an acute or subacute neurological pattern, beginning with visual defects and alimentary and social impairment, followed by a brain-stem syndrome. CT and M.R.I. will make it more easy. An earlier diagnosis could perhaps allow to discover the suspected enzymopathy responsible for Leigh's encephalomyelopathy and make clearer the relationship between Leigh's disease and encephalopathies with abnormal mitochondria. PMID- 3047834 TI - [A multi-factorial approach in the vital prognosis of spontaneous intracerebral hematoma]. AB - The prognostic assessment of a patient with intra-cerebral hemorrhage (IH) requires simultaneous appraisal of several parameters. We have attempted this with a multivariate method: discriminant analysis. We studied retrospectively 142 patients with non-operated IH, not due to vascular malformation, distributed two months after the initial event in two groups: 92 living patients and 50 dead. Discriminant analysis of 21 parameters from the initial examination and CT scan, selected five factors which best separate the two groups, since 89% of the patients were well classified. These five parameters (age, consciousness impairment, temperature, volume of the hematoma and ventricular hemorrhage) combined, give a prognostic score which gives for each patient his probability of survival or death. The validity of the proposed model was controlled on a test sample of 66 patients from another department. The possibility of giving a trustworthy spontaneous prognosis on the first day can enable the evaluation of the possible benefit from surgery, which we illustrated with a group of 23 operated patients. PMID- 3047835 TI - [Cardio-respiratory anomalies in disseminated sclerosis]. AB - A case of acute pulmonary edema without cardiac failure, infectious or toxic cause, revealing a Multiple Sclerosis (M.S.), one case of bradycardia during a bout in a known M.S. and one case of orthostatic hypotension without change of cardiac frequency, during a bout of a known M.S. are reported. The common point of these 3 cases is that during their autonomic failure, there were disorders pointing to the medulla oblongata: swallowing difficulties, rotatory nystagmus, vestibulo-cerebellar syndrome, sensory and motor defect of upper limbs. A plaque of M.S. in the medulla oblongata, particularly near the tractus solitarius could explain these cardio-pulmonary abnormalities unusual in M.S. PMID- 3047836 TI - [Thrombopenia and disseminated intravascular coagulation during treatment with heparin: 2 cases with neurological complications]. AB - We report two cases of heparin-induced thrombocytopenia (H.T.) associated with a disseminated intravascular coagulopathy (DIC). Heparin was prescribed after a cerebral infarct in the first case and after an orthopedic surgical procedure in the second case. The DIC induced neurological complications and the death of both patients. Heparin-induced thrombocytopenia is common but is exceptionally associated with neurological symptoms. Heparin-induced DIC is quite uncommon, since only 20 cases have been reported but neurological complications (focal deficits or disturbances of consciousness) are noted in about one third of the cases and the outcome is fatal in 60 percent of the cases. The treatment of heparin-induced DIC and thrombocytopenia is disappointing. Monitoring of the platelets count is warranted during heparin treatment to prevent this severe complication. PMID- 3047837 TI - [Benign cerebral angiopathies and phenylpropanolamine]. AB - Heroin, cocaine, amphetamines, sympathomimetic drugs can cause cerebral angiopathy. We report 2 patients with cerebrovascular disorders after ingestion of a nasal vasoconstrictor containing phenylpropanolamine (P.P.A.). The first patient had two acute repetitive attacks of severe headache and vomiting, occurring after a daily treatment with 180 mg of P.P.A. during 6 weeks. The second patient had an intracerebral hemorrhage, occurring some hours after taking for the first time 120 mg of P.P.A. In both cases, cerebral angiography, performed in the next week, demonstrated segmental narrowing and dilatations of medium-size intracranial arteries. None of the usual causes of cerebral vasculitis were present. The outcome was favorable and follow-up angiograms showed the disappearance of the beading pattern. P.P.A. is widely used over the counter in diet pills and stimulants. Cerebral vascular complications have been rarely reported, always hemorrhagic and often associated with cerebral vasculitis. They are unrelated to duration or dosage of treatment. The mechanism is unclear but could result from several factors: chronic or paroxystic high blood pressure, immuno-allergic vasculitis, arterial spasm, direct "toxic" effect of the P.P.A. on the arterial wall may be increased by other drugs and caffeine. PMID- 3047838 TI - [Human acquired immunodeficiency virus infection of the nervous system]. AB - Neurological manifestations of HIV infection are recognised at different phases of the evolution of the disease. During the late stages opportunistic neuro meningeal infections and tumors develop as a result of the immuno-suppression. There exist, however, various manifestations which evolve independently of the immune state, and seem to be directly related to the virus itself. One can distinguish central neuro-meningeal syndromes and peripheral syndromes at the onset, or at later stages. Their prognostic implications are uncertain, but often severe, for example in the case of the subacute dementias which, in this setting, may lead to death in several months. There are also other manifestations which may be self limited or slowly evolving in the central and peripheral nervous systems. The neuropathological changes are known for the dementias as mentioned above, though the characteristics lesions due to HIV infection per se remain controversial. The presence of the virus in the nervous system has been established by in situ hybridization techniques, immunohistochemistry and culture. Studies on CSF have also allowed virological (cultures) and immunological studies to be carried out. The physiopathological mechanisms for the apparent neurological effects of this virus remain hypothetical. A better understanding of these mechanisms should lead to a rationalisation of therapeutic strategies, in a disorder which would seem to be an early and persisting viral infection of the nervous system. PMID- 3047839 TI - [Carotid endarterectomy: long-term ultrasonic evaluation]. AB - Among 128 patients operated for atherosclerosis of the internal carotid artery, 83 (59 males, 24 females, mean age 64 years) were controlled by Doppler ultrasonography, spectral analysis and Duplex. They had been submitted to 86 endarterectomies and 5 operations for total occlusion. In 23 cases, the stenosis was asymptomatic. The delay between operation and control varied between 12 and 123 months (mean follow-up: 50.6 months). 75 patients remained asymptomatic, whereas 8 suffered TIAS. Ultrasound examinations revealed 57 normal arteries or with thickened walls (62.6 p. 100), 9 stenoses less than 50 p. 100 (10 p. 100), 14 stenoses more than 50 p. 100 (15 p. 100) among which 8 were reoperated and 11 occlusions (4 failures of desocclusion and 7 postoperative occlusions: 7.6 p. 100). Recurrent stenoses occurred more frequently in females, mean age lower than in the whole group, as has been already reported. They determined minor symptomatology (vertebro-basilar insufficiency in 2 cases, TIA in 1 case). Three postoperative occlusions were acute and gave rise to a severe neurological deficit, whereas the other ones remained generally asymptomatic. These results are compared to reported series. Ultrasonography is a technique of choice to follow endarterectomised patients. The high rate of recurrent stenoses after endarterectomy raises questions about endarterectomy. PMID- 3047840 TI - [Pigmentary orthochromatic leukodystrophy. Van Bogaert and Nyssen disease]. AB - A 43 year old woman with a strong likelihood of familial history, developed progressively a spastic tetraparesis associated with intellectual deterioration and terminal epileptic fits. She died 11 years after onset of the clinical disorders. Neuropathological study revealed an orthochromatic leukodystrophy. Macrophages and glial cells of the white matter contained a brown-yellow, autofluorescent pigment which stained positively with PAS, Perls stain for iron and Masson-Fontana. Electron microscopy showed electron dense, membrane bound intracytoplasmic lamellar inclusions with curved or straight parallel arrangement or fingerprint pattern, in white matter macrophages, astrocytes and oligodendrocytes. Cortical cells contained lipofuscin which was normal in type and amount. This suggests that the material in white matter glial cells and macrophages is ceroid pigment; however, the distribution is not that seen in ceroid-lipofuscinosis. Similar inclusions have been found in oligodentrocytes in other forms of orthochromatic leucodystrophy. Ten similar cases of pigmentary type of orthochromatic leucodystrophy have been reported previously; only one had had an ultrastructural study. PMID- 3047841 TI - [Conference at Salpetriere September 1986. Partial complex seizures with dream state. Clinical, stereo-electroencephalographic and neuropathological discussion]. PMID- 3047842 TI - A history of the Infectious Diseases Society of America. PMID- 3047843 TI - [Continuing education]. PMID- 3047844 TI - [Myopathies caused by defects of lipid and carbohydrate metabolism]. PMID- 3047845 TI - [Mitochondrial myopathies]. PMID- 3047846 TI - [EMG in myopathies]. PMID- 3047847 TI - [The contribution of biochemistry to the diagnosis of muscular diseases]. PMID- 3047849 TI - RNAO education guide 88-89. PMID- 3047848 TI - [Duchenne, Becker and limb-girdle muscular dystrophies. Problems of differential diagnosis]. PMID- 3047850 TI - Monoclonal antibodies for detection of Campylobacter pylori in biopsy smears and frozen sections. PMID- 3047851 TI - Heterogeneity of Campylobacter pylori as demonstrated by co-agglutination testing with rabbit antibodies. AB - The indirect immunofluorescence (IFL) and the co-agglutination (CoA) methods were used to study the serology of Campylobacter pylori strains isolated from patients in different countries (Sweden, Finland, Canada and Australia). Antisera were obtained from rabbits immunized with whole cell antigens. With IFL tests the highest serum titers were obtained with C. pylori strains and their homologous antisera. These tests also showed that all the tested strains contained cross reactive antigens. With the use of the CoA technique strain or type specific heat labile and heat stable antigens were demonstrated, and a provisional "seropattern" of a particular strain could be defined with selected CoA reagents. With the use of such reagents we were able to show that a patient may be infected with multiple C. pylori strains with different sets of surface antigens. The clinical and epidemiological implications of this serological heterogeneity of C. pylori are discussed. PMID- 3047852 TI - Comparison of different tests for Campylobacter pylori. PMID- 3047853 TI - Bismuth: effects on gastritis and peptic ulcer. AB - The healing properties of colloidal bismuth subcitrate (CBS) on peptic ulcer are well established and several studies have shown that healing with CBS is associated with a lower relapse rate than that produced by H2-receptor antagonists. The recent observation that CBS is effective against Campylobacter pylori has shed light on this because recent studies have shown that eradication of C. pylori by CBS leads to resolution of the associated gastritis and this may explain the low relapse rates. CBS is also effective in C. pylori positive patients with non ulcer dyspepsia (NUD) in whom clearance of these organisms from the stomach is associated with significant improvement of the associated gastritis and symptoms. PMID- 3047854 TI - The role of antibiotics in Campylobacter pylori associated peptic ulcer disease. AB - Since the recognition and first culture of Campylobacter pylori (Cp) the hypothesis of a pathogenic role has been strengthened by numerous investigations. Its close association with active chronic gastritis and even active duodenitis, and the disappearance of these pathologic conditions with antimicrobial treatment suggest a pathogenic role for Cp in active chronic gastritis. The close association of antral gastritis with duodenal ulcer (DU) suggests that Cp associated active chronic gastritis may be an important precondition for the development of DU. Therapy studies so far using either bismuth, antibiotics, or a combination of both could demonstrate that the healing of DU ulcer was closely related to the clearance of Cp and healing of gastritis. Relapse of DU was closely associated with reappearance of Cp and active chronic gastritis. At the present state the combination of bismuth salts and antibiotics has achieved the highest eradication rates of Cp and thus lowest relapse rates of DU. Antibiotics therefore seem to be an indispensable factor in the antimicrobial treatment of Cp associated diseases. PMID- 3047855 TI - Comparative trials of doxycycline versus amoxicillin, cephalexin and enoxacin in bacterial infections in chronic bronchitis and asthma. AB - The comparative efficacy of doxycycline versus amoxicillin, cephalexin, cefaclor and enoxacin was examined in four separate cross-over and blinded studies of acute bacterial bronchitis in chronic bronchitis and asthma. The efficacy of doxycycline over the eleven-year period (1975-1986) covered by these studies also was examined. Patients with acute bacterial exacerbations, defined by increased chest symptoms, increased bacteria and sputum neutrophilia, were randomly entered. When a new acute infection occurred, they were re-entered and received the other antibacterial. Response was recorded as successful or not; early rebound infections and the periods free of infection were noted. A total of 136 exacerbations were evaluated in the comparison of doxycycline with the other 4 antibacterials, and 93 exacerbations in the long-term efficacy of doxycycline. The acute success was similar for doxycycline with the other antibacterials and was superior to cefaclor. Early rebound infections occurred less frequently with doxycycline as compared to the cephalosporins and was similar versus amoxicillin and enoxacin. The infection-free period was longer after doxycycline than with the other four antimicrobials. Doxycycline maintained its efficacy to elicit a prompt response and provide a long infection-free period, but an increase of early rebound infections was noted over the eleven-year period. PMID- 3047857 TI - Urinary infection stones caused by Ureaplasma urealyticum: a review. AB - Experimental and clinical studies have been performed to determine whether Ureaplasma urealyticum has an etiological role in the development of infection stones in the urinary tract. Incubation of synthetic urine in vitro with U. urealyticum caused alkalinization of the urine and crystallization of struvite and calcium phosphate. Inoculation of U. urealyticum into rat bladders resulted in the formation of struvite stones in 84% of the rats. Furthermore, infection with U. urealyticum markedly increased the adherence of urease-induced crystals to the bladder epithelium compared to normal rat bladders, probably due to elimination of the mucous coat which covers the normal urothelium. Clinically, U. urealyticum has been cultured from voided urine and from the stone in patients operated on for renal stones. U. urealyticum was cultured in voided urine in 31 of 247 patients (13%) with metabolic stones, compared to 43 of 145 patients (30%) with infection stones (p less than 0.001). In the patients where stone cultures were performed, U. urealyticum was found in 2 of 125 patients (2%) with metabolic stones, compared to 10 of 64 patients (16%) with infection stones (p less than 0.001). These observations strongly suggest that U. urealyticum is linked to the formation of infection stones in the urinary tract. PMID- 3047856 TI - Antimicrobial agents in the treatment of periodontal diseases: special aspects on tetracycline and doxycycline. AB - Increasing evidence for the involvement of specific microorganisms in the etiology of human periodontal diseases has appeared during the last few years. The microorganisms most likely to cause periodontal disease are Actinobacillus actinomycetemcomitans, Bacteroides gingivalis, Bacteroides intermedius and Capnocytophaga species. Many of the strains are resistant to tetracycline and doxycycline. Recently penicillin-resistant B. gingivalis and B. intermedius have also been described. A. actinomycetemcomitans strains and capnocytophaga strains are sensitive to tetracycline and doxycycline. This has raised the question if periodontal diseases are possible to treat with antimicrobial agents alone or in combination with scaling of the teeth and surgery. Although numerous studies with different antimicrobial agents have been published it is still controversial if antimicrobials agents have a role in the treatment of human periodontal diseases. Comparative randomized long term studies are still needed to answer this question. PMID- 3047858 TI - Long-term effects on bacterial sensitivity patterns of preoperative antibiotic prophylaxis in colorectal surgery. AB - Since 1973, when doxycycline was introduced as preoperative prophylaxis in elective colorectal surgery at Malmo General Hospital, Sweden, there has been an unchanged and low rate (8% to 12%) of septic complications in colonic surgery. For treatment of postoperative infections, ampicillin and cefuroxime have been used since 1973 and 1980, respectively. The sensitivity of Escherichia coli to these three antibiotics used for prophylaxis and treatment was followed for five years (1981-1985). Only minor changes were observed during the period. A lower frequency of antibiotic resistance was found in bacterial strains isolated peroperatively than in strains isolated postoperatively after colorectal surgery or from infections in other patients. Considering the low frequency of postoperative infectious complications and the low frequency of antibiotic resistance in peroperative isolates, doxycycline still remains an alternative for prophylaxis in bowel surgery. PMID- 3047859 TI - [Feline malignant histiocytosis and lysozyme detection]. PMID- 3047860 TI - [Clinical and serological diagnosis of ehrlichiosis in dogs in Switzerland]. PMID- 3047861 TI - [Ultrasound scanning of thickening of the gallbladder wall and its diagnostic significance in critical care patients]. AB - In a prospective study including 398 intensive-care patients, we analysed the ultrasonographic relevance of a gallbladder wall thickening. In 24 of 398 (6%) cases a wall thickening was found that could be differentiated into two types of walls. In 20.8% (5/24) the gallbladder wall thickening was an expression of an acute cholecystitis. In further differential diagnosis of a gallbladder wall thickening pathological states with hypoalbuminaemia occupy the prime position. The pathological mechanism has not yet been completely clarified. Hypoalbuminaemia without inflammatory alterations of the organ was found in 37.5% of the cases with gallbladder wall thickening. In 41.6% of the patients the wall thickening had to be taken as a non-specific ultrasonographic sign. On the whole, therefore, gallbladder wall thickening has thus to be valued primarily as non specific sign. In a few cases only, it implies an autonomous affection of the gallbladder. PMID- 3047862 TI - [Pathologic wall changes in the gallbladder in patients with gallstones and in healthy probands. Imaging using high frequency annular array sector transducers]. AB - 2154 computerized ultrasound diagnoses of 1623 patients were evaluated retrospectively from January 1st to October 31st 1987. The frequency of pathologic gallbladder wall changes in patients with stones and normal controls was examined applying 5 MHz annular array sector transducer. The occurrence of polyps, septae, sludge, kinking, carcinoma and adenomyomatosis was noted separately. The prevalence of polyps and septae in gallstone patients was increased fourfold if compared to the total number of examined patients. Sludge was found in 5.9% of gallstone patients compared to 2.2% in normal controls. Carcinoma and adenomyomatosis were present only in cases of cholelithiasis; the prevalence of these pathologic wall changes is obviously rare. The importance of polyps and septae in biliary lithogenicity is discussed. PMID- 3047863 TI - [Streptococcal abscesses of the liver: diagnosis and therapy using ultrasound controlled puncture]. AB - Three male patients without accompanying diseases, 27, 47, and 52 years of age, were admitted with fever of unknown origin. Focal liver disease was seen on ultrasound and the diagnosis of an abscess was confirmed by sonographically guided fine-needle puncture. Microaerophilic streptococci were found in two cases, streptococcus viridans once. Two patients had cryptogenic abscesses; one had previously undergone surgery for a rectal abscess. All three patients were cured of their condition by appropriate antibiotic coverage and evacuation of the abscess by needle aspiration. PMID- 3047864 TI - [Risks of fine needle puncture--results of a survey in West Germany(German Society of Ultrasound in Medicine survey)]. AB - The rate of complications in fine needle puncture was ascertained by an inquiry conducted among the members of the German Association of Ultrasound in Medicine (DEGUM). 337 complications had been registered in a total of 66,397 punctures, corresponding to a percentage of only 0.51%. 294 of these were mild, i.e. not requiring therapy (0.44%), 38 severe (0.057%) and 5 complications were fatal and resulted in death (0.0075%). Differences were slight in respect of the frequency of complications with reference to puncture technique and type of needle employed. Most complications were seen in fine needle puncture for obtaining histological material (0.67%) (cutting biopsy needles), compared with 0.54% in therapeutic puncture and 0.46% for obtaining cytological material. Among the puncture techniques, the use of a puncture transducer resulted in more complications (0.72%) compared with sonographically guided puncture (0.59%) and puncture in full view with 0.43%. Punctures with/fatal outcome had been conducted in 4 out 5 cases with so-called cutting biopsy needles. Results are identical with those obtained in the U.S.A. on a comparative basis. Compared with punctures with so-called coarse needles, fine needle puncture results in fewer complications on account of the smaller needle caliber. As with every intensive method, however, complications must be taken into account even with fine needle puncture. PMID- 3047865 TI - [Percutaneous biopsy technic. Aspiration fine needle and cutting biopsy cannula in an experimental comparison]. AB - Guided percutaneous biopsy for cytological or histological diagnosis is well tried. Various types of needles and calibres are available. Only few studies have been performed comparing different needle configurations. Fine needles and cutting needles were evaluated quantitatively and qualitatively in an experimental study obtaining specimens of resected tumours. Weight measurement and preservation of the material were analysed. Material for cytological diagnosis was obtained with all types of needles. Increasing needle size resulted in higher yield of diagnostic tissue. Small-bore cutting needles having an acute bevel angle provide better results in soft or semiliquid tissue. It is not necessary to increase the needle size over 18 gauge. PMID- 3047866 TI - [Ultrasound controlled percutaneous transhepatic cholangiography]. AB - A method of performing transcutaneous cholangiography under sonographic control is presented. This method makes it possible to enter the distended intrahepatic bile ducts accurately. The sonographic control enables the needle to be inserted into the superficial bile ducts. The method reduces liver traumatisation. Under the control of sonography the procedure is safer, and there are fewer complications. PMID- 3047867 TI - [Ultrasound pinpointing of foreign bodies. An in vitro study]. AB - A study of ultrasonographic guided pinpointing of foreign bodies inserted into pork muscle tissue is presented. The optimal results were obtained using two needles inserted at an angle of approximately 90 degrees to each other and crossing just underneath the foreign body. This method facilitates surgical removal of foreign bodies considerably. PMID- 3047869 TI - FDA looks to speed up drug approval process. PMID- 3047868 TI - [ Aneurysma spurium of the pancreaticoduodenal artery]. AB - A predominantly cystic process of the upper abdominal region of a 61 year old female patient was punctured via the fine-needle technique. The clinical assumption had been a lymphoma. Bright red blood was aspirated. The Doppler sonographic measurement revealed an arterial pulsating aneurysmatic process, and this was subsequently proved by selective angiography and, finally, by surgical treatment. PMID- 3047870 TI - A heresy in evolutionary biology. PMID- 3047871 TI - A chemoattractant receptor controls development in Dictyostelium discoideum. AB - During the early stages of its developmental program, Dictyostelium discoideum expresses cell surface cyclic adenosine monophosphate (cyclic AMP) receptors. It has been suggested that these receptors coordinate the aggregation of individual cells into a multicellular organism and regulate the expression of a large number of developmentally regulated genes. The complementary DNA (cDNA) for the cyclic AMP receptor has now been cloned from lambda gt-11 libraries by screening with specific antiserum. The 2-kilobase messenger RNA (mRNA) that encodes the receptor is undetectable in growing cells, rises to a maximum at 3 to 4 hours of development, and then declines. In vitro transcribed complementary RNA, when hybridized to cellular mRNA, specifically arrests in vitro translation of the receptor polypeptide. When the cDNA is expressed in Dictyostelium cells, the undifferentiated cells specifically bind cyclic AMP. Cell lines transformed with a vector that expresses complementary mRNA (antisense) do not express the cyclic AMP receptor protein. These cells fail to enter the aggregation stage of development during starvation, whereas control and wild-type cells aggregate and complete the developmental program within 24 hours. The phenotype of the antisense transformants suggests that the cyclic AMP receptor is essential for development. The deduced amino acid sequence of the receptor reveals a high percentage of hydrophobic residues grouped in seven domains, similar to the rhodopsins and other receptors believed to interact with G proteins. It shares amino acid sequence identity and is immunologically cross-reactive with bovine rhodopsin. A model is proposed in which the cyclic AMP receptor crosses the bilayer seven times with a serine-rich cytoplasmic carboxyl terminus, the proposed site of ligand-induced receptor phosphorylation. PMID- 3047873 TI - Virus-specific splicing inhibitor in extracts from cells infected with HIV-1. AB - Human immunodeficiency virus type 1 (HIV-1), in contrast with most other retroviruses, encodes trans-regulatory proteins for virus gene expression. It is shown in this study, by means of an in vitro splicing system, that nuclear extracts obtained from cells infected with HIV-1 contain a factor (or factors) that specifically inhibits splicing of a synthetic SP6/HIV pre-messenger RNA (pre mRNA)-containing donor and acceptor splice sites in the coding region for the envelope protein. It is also shown that the SP6/HIV pre-mRNA is not capable of assembly in a ribonucleoprotein complex, spliceosome, in extracts from infected cells. These findings raise the possibility that specific inhibition of pre-mRNA splicing in the envelope protein coding region by HIV-1 trans-regulatory factors might be one control mechanism for efficient production of structural viral proteins and virion assembly. PMID- 3047872 TI - Zinc-dependent structure of a single-finger domain of yeast ADR1. AB - In the proposed "zinc finger" DNA-binding motif, each repeat unit binds a zinc metal ion through invariant Cys and His residues and this drives the folding of each 30-residue unit into an independent nucleic acid-binding domain. To obtain structural information, we synthesized single and double zinc finger peptides from the yeast transcription activator ADR1, and assessed the metal-binding and DNA-binding properties of these peptides, as well as the solution structure of the metal-stabilized domains, with the use of a variety of spectroscopic techniques. A single zinc finger can exist as an independent structure sufficient for zinc-dependent DNA binding. An experimentally determined model of the single finger is proposed that is consistent with circular dichroism, one- and two dimensional nuclear magnetic resonance, and visual spectroscopy of the single finger peptide reconstituted in the presence of zinc. PMID- 3047874 TI - Symmetrical erosive peripheral polyarthritis in the Late Archaic Period of Alabama. AB - Rheumatoid arthritis was first described unambiguously in 1800, but its etiology and historical origins are still obscure. Definite rheumatoid arthritis has not been demonstrated in pre-19th century Old World skeletal remains. Six individuals who lived 3000 to 5000 years ago in northwestern Alabama and present erosive polyarthritis characteristic of rheumatoid arthritis are described. The diagnosis raises the possibility that rheumatoid arthritis can be associated with a New World pathogen or allergen. PMID- 3047875 TI - Cytokines alter production of HIV-1 from primary mononuclear phagocytes. AB - Some strains of human immunodeficiency virus type 1 (HIV-1) can infect primary monocytes and monocyte-derived macrophages in vitro. In this report, the effect of cytokines on the production of one of these strains that shows a tropism for mononuclear phagocytes, designated HIV-1JR-FL, was studied. Primary peripheral blood mononuclear phagocytes infected with HIV-1JR-FL were treated with the hematopoietic factors: granulocyte colony-stimulating factor (G-CSF), granulocyte macrophage colony-stimulating factor (GM-CSF), interleukin-3 (IL-3), macrophage colony-stimulating factor (M-CSF), and gamma-interferon (gamma-IFN). The M-CSF, GM-CSF, IL-3, and gamma-IFN were able to alter HIV-1 production under different conditions. PMID- 3047876 TI - [Antibiotics in 1988]. PMID- 3047877 TI - Carbamazepine in the treatment of phantom limb pain. AB - The successful use of carbamazepine in treating a case of severe phantom limb pain prompted me to review this condition and its experiential treatment with carbamazepine, as well as the drug's possible mechanism of action on phantom limb pain. Experience to date indicates the need for controlled double-blind crossover studies to test the therapeutic validity of carbamazepine. PMID- 3047878 TI - Clonazepam, haloperidol, and clonidine in tic disorders. AB - We reviewed the treatment outcomes of 81 patients with multifocal tic disorders followed in our movement disorders clinic. From the three drugs used, rank orders of effectiveness were haloperidol, clonazepam, and clonidine. Because of the risk of tardive dyskinesia, we suggest first a trial with clonazepam, later combined with clonidine if clonazepam alone is not effective. Haloperidol should be reserved for the most disabling cases, and given only after trials with both clonazepam and clonidine have failed. PMID- 3047879 TI - Strangulated obturator hernia: can mortality be reduced? AB - We describe two patients with strangulated obturator hernia to enhance clinical awareness of the varied presentations of this uncommon hernia, which occurs especially in elderly women with either recurrent abdominal pain or partial intestinal obstruction, a positive Howship-Romberg sign, and an absent thigh adductor reflex. Early laparotomy for unexplained bowel obstruction is essential to avoid the complications associated with strangulated obturator hernia. PMID- 3047880 TI - Hematologic effects of heavy metal poisoning. AB - Heavy metal poisoning can cause a variety of hematologic disorders. Exposure to heavy metals is ubiquitous in the industrial environment and must be considered in the differential diagnosis of many types of anemia. The heavy metals most commonly associated with hematologic toxicity are arsenic and its derivative arsine, copper, gold, lead, and zinc. A few distinctive clinical features characterize the hematologic manifestations of many occult heavy metal poisonings. These features have a limited differential diagnosis. A knowledge of these clinical features can assist the astute clinician in making the correct diagnosis. PMID- 3047881 TI - Efficacy of the lateral decubitus position in preventing pneumothorax after needle biopsy of the lung. AB - We evaluated the efficacy of the lateral decubitus position in preventing the development of pneumothorax after percutaneous needle biopsy of a well defined lung lesion. Fifty patients had such a procedure and were immediately placed in the lateral decubitus position for ten minutes, with the biopsied lung below. A chest x-ray film was obtained after the ten minute period, four to six hours later, and on the following morning. The number of pneumothoraces and the type of treatment needed in each case were recorded. Results in these 50 patients were compared to those previously obtained in 132 patients who had had an identical procedure and follow-up during the preceding four years. In all patients included as controls, we did the biopsy using an identical technique and in accordance with a specific written protocol. Both groups of patients were closely comparable in age and sex distribution, baseline values for arterial blood gases, results of prebiopsy tests of pulmonary function, and type, size, location, and distance from the pleural surface of the pulmonary lesions. A postbiopsy pneumothorax developed in 21 of 50 (42%) study patients, and in 46 of 132 (35%) controls (P not significant). Specific therapy was needed in ten (48%) and 26 (56%) of these cases, respectively (P = NS). We conclude that placing patients in the lateral decubitus position is not effective in significantly reducing the incidence of postbiopsy pneumothorax or the proportion of patients who will require specific therapy for this complication. PMID- 3047882 TI - Preventive cardiology: what is the value of antiplatelet agents and fish oils? PMID- 3047883 TI - The caduceus as a medical emblem: heritage or heresy? PMID- 3047885 TI - New horizons in chronic granulocytic leukemia. PMID- 3047884 TI - Acute pancreatitis caused by Legionella pneumophila. AB - A 57-year-old man had Legionella pneumonia and acute pancreatitis. The diagnosis of legionellosis was established by both culture and significant rise in antibody titer to Legionella pneumophila. The pancreatitis was diagnosed by elevated amylase and lipase values, peaking on the fourth hospital day, roentgenologic findings, and a clinical picture compatible with the disease. The patient was not an alcohol consumer and had had no previous pancreatic disease. We conclude that acute pancreatitis can be a manifestation of legionellosis. PMID- 3047886 TI - Methylphenidate v. placebo--a randomised double-blind crossover study in children with the attention deficit disorder. AB - Fourteen pupils of the New Hope School in Pretoria who were considered to be responsive to methylphenidate were randomised to receive either the active drug or placebo in a double-blind trial for each of 4 weeks. Their behaviour was monitored by parents and teachers, who were required to complete a questionnaire (Conner's Abbreviated Teachers' Rating Scale) three times per week. After analysis of the data, only 2 children were identified as methylphenidate responders and 1 as a probable responder. One child showed significant deterioration on the drug while another showed deterioration that approached significance. The remaining 9 exhibited no significant response to methylphenidate. PMID- 3047888 TI - Hysterectomy for septic abortion--is bilateral salpingo-oophorectomy necessary? AB - Ovarian conservation at the time of hysterectomy for complicated septic abortion is important in this young population group. In a retrospective study, the histological evaluation of the ovaries of 25 patients were compared with the macroscopic description in the operation reports. In 72.3% of the ovaries examined there was no infection. None of the ovaries described clinically as normal at laparotomy showed histological signs of infection. The clinical assessment of infected ovaries was false-positive in 40% of cases but there was no false-negative decision-making. It is concluded that ovaries which appear normal at hysterectomy for septic abortion should be conserved. PMID- 3047887 TI - Effect of indapamide on serum and red cell cations, with and without magnesium supplementation, in subjects with mild hypertension. AB - Unlike some thiazide diuretics, indapamide--a non-thiazide chlorosulphonamide derivative--has been shown to have a magnesium-sparing effect in normotensive subjects. This effect has not been studied in hypertensive subjects. In a randomised double-blind trial indapamide 2,5 mg and placebo were given daily to a group of elderly patients with mild hypertension, with and without supplemental magnesium chloride. Blood pressure and serum and red blood cell cations were measured. The significant antihypertensive effect of indapamide was confirmed. There was no effect of indapamide on serum and red cell magnesium concentrations compared with placebo, both with and without magnesium supplementation. Indapamide induced hypokalaemia with a shift of sodium into the red cells. In this group of elderly hypertensive subjects indapamide induced potassium but not magnesium loss. PMID- 3047890 TI - Mamre--history and development. AB - Mamre's history spans the period of Khoikhoi habitation of the area to the present-day village. In 1808 a Moravian mission station was established there. The church remains an important feature of the modernising community, which is currently under secular control. PMID- 3047889 TI - The 1918 influenza epidemic in Mamre. AB - The 1918 influenza epidemic was a historic, health and demographic landmark in South Africa. In Mamre the mortality rate was 39/1,000, which was similar to the rate in Cape Town. The male/female ratio was 1,33, with males between the ages of 20 and 40 years accounting for 60% of deaths. PMID- 3047891 TI - The prophylaxis of psycho-somatic diseases of the workers employed in continuous work flow lines and mass production car plants. PMID- 3047895 TI - The pre-Halstedian and post-Halstedian history of the surgical rubber glove. PMID- 3047893 TI - A randomized trial of tamponade or sclerotherapy as immediate treatment for bleeding esophageal varices. AB - Sengstaken-Blakemore tamponade is used for the initial control of bleeding esophageal varices (BEV), although it is known to be potentially dangerous. Sclerotherapy has been shown to be effective in the treatment of BEV. This trial has been designed to evaluate comparatively the effectiveness of both procedures in the initial control of the hemorrhage. Forty-three patients with BEV were included in the trial. Twenty patients (group SB) were treated by tamponade. Twenty-three patients (group ST) were treated by sclerotherapy by means of a simple technique. During the first 24 hours, hemostasis was obtained in 16 of the SB patients and in all 23 of the ST patients (p less than 0.05). At seven days, nine SB patients and 19 ST patients were free of hemorrhagic relapse (p less than 0.05). By stratifying in relation to hepatic failure, the difference was greater (p less than 0.005) if patients with Child's A classification were excluded. It is concluded that sclerotherapy should be undertaken in almost every instance at the same moment that diagnosis is made, bypassing the intermediate step of tamponade. PMID- 3047894 TI - A review of a Plastibell device in neonatal circumcision in 2,000 instances. AB - The Plastibell technique is a simple, satisfactory and easily learned method of circumcision. It is important to stress the practicality of this method in the neonatal period, thus reducing both emotional trauma and financial cost of admission at a later date. PMID- 3047892 TI - Progress in acute pancreatitis. AB - Pancreatic necrosis and sepsis are the major causes of death in instances of acute pancreatitis. No widely accepted definition of these conditions in individuals exists, and, yet, accurate differentiation is mandatory for effective therapy. A series of operational definitions conforming to known clinopathologic factors are proposed for the necrotizing septic complications of acute pancreatitis. These complications, as distinguished from acute interstitial pancreatitis, are fat sequestra, pancreatic necrosis, infected pancreatic necrosis, pancreatic abscess and acute pseudocyst. Imprecise definitions of these complications of necrotizing pancreatitis make inter-institutional comparisons of previously identified data dubious. PMID- 3047896 TI - [Radiation therapy of nasopharyngeal carcinoma. Retrospective study for the assessment of long-term results]. AB - During the years from 1967 to 1986, 90 patients with nasopharyngeal carcinomas were irradiated at the Radiologic Hospital of Bonn University. The median overall survival time was 4.3 years which corresponded to 23% of the statistical life expectancy of the patients. 66% were still alive after two years and 46.5% after five years. Compared to the overall group, the prognosis was significantly worse in case of highly differentiated, keratinizing squamous cell carcinomas, penetration of the primary tumor into the base of the skull, or certain symptoms as ophthalmo-neurologic troubles, headaches, loss of body weight, night sweat, or fever. After partial tumor excision performed prior to irradiation or complete remission following to radiotherapy, the therapy results were better than the average with median survival times of about eleven years. PMID- 3047898 TI - [Optimization of combined brachy-teletherapy by the modification technic]. AB - A method is presented for combined brachy-teletherapy by which the dose distribution of percutaneous irradiation is adapted to the distribution of short distance irradiation with 192iridium by means of the so-called "field integrated dose modification". Insufficient or excessive doses within the area of superposition of both therapy modalities can so be avoided. Moreover, the therapist can define zones of deviation from the reference dose, if such a dose heterogeneity seems desirable in the individual case. The combined irradiation of prostata and cervix is given as an example to describe in detail the technical, clinical, and radiobiological aspects of this method. PMID- 3047897 TI - [The development of radiation caries after high doses of irradiation]. AB - 39 patients, who were irradiated with doses of 50 to 70 Gy for ENT-tumors over a period of 3.5 months to three years prior to the examination, showed a rapidly progressing caries of the teeth inside the target volume. The teeth outside the target volume developed a caries of less extent. Radiation induced xerostomia, effects of the irradiation of the soft tissues, nutrition habits and hygiene are discussed as causes for the damage of the teeth. PMID- 3047899 TI - Why study the epidemiology of asthma? PMID- 3047901 TI - Cardiac tamponade due to pneumopericardium. AB - Pneumopericardium is rare in acute asthma and cardiac tamponade has not been reported. The case is reported of a 20 year old asthmatic patient in whom assisted ventilation and high airway pressures resulted in tension pneumopericardium with clinical signs of cardiac tamponade that were relieved by pericardial aspiration. PMID- 3047900 TI - Efficacy of "mucoregulatory" agents in Young's syndrome. AB - Eight patients with Young's syndrome were treated with four "mucoregulatory" agents for eight weeks in a randomised, open crossover study. There was no improvement in tracheobronchial clearance, pulmonary function, or sperm count. PMID- 3047902 TI - Atraumatic suppurative mediastinitis and purulent pericarditis due to Eikenella corrodens. AB - Atraumatic suppurative mediastinitis is an uncommon infection. A case with an associated purulent pericarditis caused by Eikenella corrodens is reported. PMID- 3047903 TI - Intravascular missile: apparent retrograde course from the left ventricle. AB - An air gun pellet was found in the right superior pulmonary vein after penetrating the left ventricle of a 14 year old boy. This apparent retrograde movement in the left side of the heart has not been reported previously. PMID- 3047904 TI - [Endogenous or "digoxin-like" digitalis compounds. Physiopathological, analytical and pharmacokinetic impact]. PMID- 3047905 TI - [Thrombopenia induced by heparin. Symptoms, diagnosis and frequency]. PMID- 3047906 TI - [Tumor necrosis factor]. PMID- 3047907 TI - [General and special problems concerning the approval and re-evaluation of antimicrobial agents for use in agriculturally useful animals]. AB - Problems associated with registration and reevaluation of drugs are discussed in this paper. In particular it deals with difficulties resulting from toleration of residue concentrations in edible tissues and from the deduction of withdrawal periods. Possible consequences when applying identical criteria for registration and reevaluation are outlined. The necessity to involve the whole veterinary profession, when setting criteria for determination, presentation and evaluation of clinical efficacy, is stressed. PMID- 3047908 TI - [Is a repeat treatment of antibiotic therapy necessary?]. AB - Interrelations between host, bacterium and antibiotic compound in antibiotic therapy of bacterial infections are analysed. Some methods for the evaluation of antibacterial activity of antibiotics in vitro are described and their limits, in particular of the widely used MIC tests, are demonstrated. Data from trials comparing antibiotic applications once and twice a day as well as discussions in chemotherapy, concerning the applicability of in vitro results to therapy, are used to try to find practicable guidelines for the repetition of antibiotic therapy. PMID- 3047909 TI - [The pharmacokinetics of common chemotherapeutic drugs used in veterinary medicine]. AB - The present paper summarizes the pharmacokinetics of chemotherapeutic agents commonly used in controlling bacterial disease in animals. Data on absorption, distribution and excretion of chemotherapeutic agents are given in form of tables. It is explained that pharmacokinetic parameters of antimicrobial chemotherapy among others are influenced by the drug's pharmaceutical formulation, its route of administration, its biotransformation and also by the age of the patients. There are given hints for handling the drug in order to achieve optimal effects. PMID- 3047910 TI - [The principle of "balanced anesthesia" in high risk canine patients]. AB - The concept of balanced anaesthesia aims at an anaesthetic method for high risk patients by combination of different anaesthetic drugs. The anaesthetics and muscle-relaxants have to work in a way as to potentiate their effects and to reduce the side effects as well as the applied dosage. With this sensible regime the circulation and the metabolic energy should be less burdened. For the introduction of balanced anaesthesia, a combination of benzodiazepines (relaxing and sedative), morphine (analgetic and sedative) and quickly metabolizing hypnotics of the imidazol group (hypnotic and relaxing) is used. A fentanyl-drip infusion continues and deepens the analgesia. After endotracheal intubation with the help of the carrier-gas nitrous oxide (weakly analgetic) and oxygen (2:1) a low concentration of volatile inhalation anaesthetics (weakly hypnotic) is added. Under artificial respiration low doses of competitive antidepolarizing neuromuscular blockers help to potentiate and reduce drug amounts of other anaesthetics especially of that for inhalation. At the end of balanced anaesthesia an antagonization of benzodiazepines, morphines and competitive neuromuscular blockers is recommended for emergency situations only because of possibly occurring changes of cardiovascular parameters. PMID- 3047911 TI - [Antibiotic treatment of diarrhea in dogs and cats]. AB - The most important reason for diarrhea in dogs and cats are dietary errors. In addition there is a multitude of other influences which may lead to diarrhea. Bacteria play a minor role in diarrhea-associated disease. The so-called primary bacterial intestinal infections occur rarely as it is shown in the examinations presented. Except for the situation in which a specific bacterial activator is the cause of continuous diarrhea in general the administration of antibiotics is ineffective. The treatment of diarrhea should rather consist of food deprivation, oral or parenteral rehydration, use of antisecretory medicaments and of adsorbents, diet and, in case of spasms, administration of anti-cholinergics. PMID- 3047912 TI - [Discovery of an atypical missile in the vascular bed]. PMID- 3047913 TI - Management of selected intraocular lens complications. PMID- 3047914 TI - Decentration of a posterior chamber lens. PMID- 3047915 TI - Current surgical management of aphakia. PMID- 3047916 TI - Surgical correction of postoperative astigmatism. PMID- 3047917 TI - Sulcus versus bag fixation of intraocular lenses. PMID- 3047918 TI - Incision and closure: astigmatism control. PMID- 3047919 TI - Risk of HIV infection in recipients of untested blood from donors now anti-HIV positive. AB - Recipients of untested blood from donors who at a subsequent donation were positive for HIV antibody by enzyme immunoassay (EIA) were evaluated, whether the result on Western blot (WB) assay was negative (EIA+/WB-) or positive (EIA+/WB+). For 109 EIA+/WB- donors, 78 recipients were tested for HIV antibody, and 3 (4%) were positive. Two of the three anti-HIV-positive recipients had clotting disorders, and the other had been massively transfused; in each of these three cases, subsequent test data exonerated the EIA+/WB- donor. For 101 current EIA+/WB+ donors, 35 recipients were tested for HIV antibody, and 13 (37%) were positive. For donors subsequently found to be EIA+/WB+, the rate of isolation of HIV was the same whether the recipients were anti-HIV-positive or anti-HIV negative (each, 5/6). While recipients of blood from donors subsequently found to be EIA+/WB+ were at substantial risk for HIV infection, regardless of the donor's subsequent HIV culture result, risk of HIV infection was not demonstrated for recipients of blood from donors later found to be EIA+/WB-. PMID- 3047920 TI - Iron supplementation in female blood donors deferred by copper sulfate screening. AB - Female blood donors with low hematocrit levels detected by copper sulfate screening were selected randomly to receive either 75 mg of iron per day, as ferrous gluconate, or a calcium phosphate placebo. Their ferritin, serum iron, total iron-binding capacity, zinc protoporphyrin, and hemoglobin values, as well as their suitability to donate blood, were determined initially (Visit 1) and at four follow-up visits (Visits 2-5). By the second visit, the serum ferritin and iron values of donors receiving iron supplementation differed significantly from those of donors receiving placebo. By the fifth visit, a less marked but significant increase in hemoglobin had occurred in the iron group, but not in the placebo group. At no time was there a significant difference between the groups' suitability to donate blood, with each group donating at almost half of their visits. The authors conclude that iron supplementation at this dose level in deferred female blood donors improves their iron status and hemoglobin levels, but does not significantly increase their suitability to donate blood as compared with the suitability of placebo-treated donors. PMID- 3047921 TI - Blood component use in orthotopic liver transplantation. AB - Blood component use during orthotopic liver transplantation (OLT) was evaluated after an initial 23-month experience with 37 consecutive transplant procedures. Blood component support of OLTs in 24 adult and 13 pediatric patients was reviewed. Adult procedures required intraoperatively a mean of 24.5 units of red cells (RBCs), 38.7 units of fresh-frozen plasma (FFP), 26.1 random-donor platelets (RDP), and 12.2 units of cryoprecipitate (Cryo); pediatric procedures required 4.8 units of RBCs, 5.8 of FFP, 3.9 of RDP, and 1.2 of Cryo. RBC salvage constituted 17 percent of the RBCs transfused intraoperatively. Intraoperative support in adult and pediatric OLT patients accounted for the majority of the total components required for the entire hospital stay. OLT blood component use constituted 1.3, 7.0, 3.6, and 8.1 percent of hospital-wide use of RBC, FFP, RDP, and Cryo, respectively, during the period of the study. PMID- 3047922 TI - Noninvasive assessment of treatment of cardiac allograft rejection with indium 111-labeled lymphocytes. AB - We have shown previously that cardiac allograft rejection can be detected noninvasively with gamma scintigraphy after administration of indium-111 (111In) labeled lymphocytes. To determine whether this technique could be used to monitor salvage immunosuppressive therapy in reversing rejection, 5 dogs were studied after thoracic heterotopic cardiac transplantation. Initial postoperative immunosuppression was maintained with cyclosporine (10-20 mg/kg/day) and prednisone (1 mg/kg/day) for 7 days after transplantation and then discontinued. Scintigraphy after administration of labeled lymphocytes was performed during initial immunosuppression and every 3 days after its termination. Endomyocardial biopsies were obtained on each day scintigraphy was performed. Once scintigraphic criteria for rejection were met (111In-lymphocyte uptake greater than mean +/- 2SD of normal myocardium), animals were treated with high dose methylprednisolone and cyclosporine. Myocardial 111In-lymphocyte activity compared with that in blood was 0.7 +/- 0.8 during initial immunosuppression, increased to 5.7 +/- 3.5 after termination of therapy (P less than 0.01), and diminished with salvage immunosuppressive therapy to 0.5 +/- 0.8 (P = NS compared with native hearts or allografts during initial immunosuppression). Scintigraphy accurately predicted all but one episode of biopsy-documented rejection and accurately detected reversal of rejection during salvage. Thus, scintigraphy with 111In-labeled lymphocytes should facilitate noninvasive monitoring of antirejection therapy in patients. PMID- 3047923 TI - Metabolic changes preceding functional and morphologic indices of rejection in heterotopic cardiac allografts. A 31P nuclear magnetic resonance study. AB - Eight beagles receiving heterotopic (cervical) cardiac allografts from outbred donors were evaluated by serial 31P NMR, septal endocardial biopsy, and left ventricular pressure measurements for signs of rejection. Early postoperative myocardial energy levels, as assessed by ratios of phosphocreatine to inorganic phosphate (PCr/Pi) and phosphocreatine to beta-ATP (PCr/B-ATP), were acceptable in all recipients. In these nonimmunosuppressed animals, the mean ratios of PCr/Pi and PCr/B-ATP progressively decreased, with a greater than 25% reduction noted by postoperative day two and greater than 50% reduction by day three. In sharp contrast, left ventricular end-diastolic pressures remained stable and at baseline levels for the first three postoperative days, and only then markedly increased. Likewise, histologic evidence of rejection did not become prominent until postoperative day four. These results suggest that metabolic abnormalities significantly precede either functional or histologic changes in rejecting allografts. The early detection of these metabolic changes by 31P NMR appears to have important potential for the noninvasive diagnosis of cardiac allograft rejection. PMID- 3047924 TI - The significance of a donor-specific vessel crossmatch in renal transplantation. AB - Very early graft rejection is usually attributed to pretransplant sensitization to HLA antigens represented on the lymphocyte. However, it is possible that such rejection episodes are secondary to antibody to a transplant-relevant system that is not represented on the lymphocyte but is represented within the allograft. This study suggests that sensitization to antigens on the VEC is detrimental to allograft success and can occur in the absence of sensitization to HLA antigens on the T or B cell or monocyte. When antibody to donor VEC is not present pretransplant, almost all patients (95%) will have a very benign posttransplant clinical course. When anti VEC antibody is present, early graft rejection or severe rejection episodes occur with a very high frequency (80%) in the nondiabetic patient. These observations, therefore, suggest that the pretransplant presence of antibody to VEC is detrimental to graft survival, and that such antibody can develop in the absence of anti HLA antibody. While the presence of such antibody is not an absolute contraindication to transplantation it does appear to represent a significant risk factor for immunological graft loss. PMID- 3047925 TI - Characterization of kidney cell-specific, non-major histocompatibility complex alloantigen using antibodies eluted from rejected human renal allografts. AB - Previous studies from our laboratories (1) have indicated that eluates prepared from rejected kidneys have antibodies not only reactive to MHC class I and class II antigens but also to antigens unique for monocytes, endothelial cells, and kidney cells. To further define the serological and biochemical nature of antigens detected by the antibodies eluted from rejected kidneys, 13 eluates prepared from rejected renal allografts and two eluates from normal kidneys were absorbed with pooled platelets, normal splenic leukocytes and/or B cells from chronic lymphocytic leukemia patients. All of the eluates failed to react with normal T and B lymphocytes by microcytotoxicity and immunofluorescence assays. However, the eluates from rejected kidneys, but not normal kidneys, reacted with peripheral blood monocytes, endothelial cells cultured from umbilical cord as well as primary kidney cells. Detailed immunohistochemical analysis of frozen kidney sections showed that the eluates from rejected kidneys reacted with structures of the cortex while sparing the structures in the medullary region. All eluates bound to the glomerulus with intense fluorescence, but not to the mesangium and Bowman's capsule, while binding to interstitial capillaries, venous endothelium, and tubular epithelium varied. More significantly, none of the eluates reacted with frozen sections of the liver, spleen, or lymph node including the endothelial lining of blood vessels in these organs. Thus, the data indicate that the eluates prepared from rejected kidneys are recognizing organ specific antigens expressed on the kidney cells. To identify the alloantigen(s) recognized by the eluted antibodies, an immunoblot analysis was performed against phase separated membrane proteins isolated from cortical kidney cells solubilized in 2X precondensed Triton X-114. A protein of Mr. 90,000-100,000 was identified as the target non-MHC antigen to which the eluates prepared from rejected allografts were reactive. Furthermore, our results suggest a possible polymorphism among these antigens. PMID- 3047926 TI - Impaired glucose tolerance in cyclosporine-prednisolone-treated renal graft recipients. AB - In 33 renal transplant patients, an intravenous glucose tolerance test (IVGTT) was performed, and fasting plasma C-peptide concentrations analyzed. Nineteen of the patients were on treatment with cyclosporine-prednisolone (CsA-Po), and 14 were treated with azathioprine-prednisolone (Aza-Po). In the Aza-Po group, the K values at IVGTT were normal in 13/14, but in the CsA-Po group they were only normal in 9/19 (P = 0.02). The fasting C-peptide levels were significantly higher in the CsA-Po group (P less than 0.001). within this group, there was no correlation between C-peptide level and serum-creatinine concentration, i.e. retention of C-peptide due to decreased renal function as judged by serum creatinine level was not suspected. Intrinsic prednisolone clearance was determined in the CsA-Po patients and was found to be lower than that previously described in Aza-Po patients. However, between those CsA-Po-treated patients with a pathologic K-value at IVGTT and those with a normal K-value there was no difference in prednisolone clearance, fasting C-peptide levels, CsA dose, or serum-creatinine concentrations. The pathophysiology of the cyclosporine-induced glucose intolerance is uncertain, and increased insulin resistance is possible. PMID- 3047928 TI - Indocyanine green clearance in acute rejection after liver transplantation. AB - Previous animals studies have demonstrated a fall in liver blood flow associated with acute rejection after liver transplantation. To study the relationship between rejection and liver blood flow in humans after liver transplantation indocyanine green (ICG) clearance was measured serially in 7 patients with clinical and histological features of acute rejection. There was a consistent pattern of satisfactory initial ICG clearance that fell in association with acute rejection and rose with successful treatment of the episode of rejection. In one patient there was no improvement in ICG clearance after treatment with additional immunosuppression, and she subsequently required retransplantation for chronic rejection. The volume of distribution of ICG was also estimated and fell considerably during the first weeks after transplantation. These results show that rejection is associated with a reduction in ICG clearance that may be due to a fall in liver blood flow, and that graft ischemia and rejection may therefore be interrelated, and important in one another's etiology. PMID- 3047929 TI - The effect of liver transplantation in a 13-year-old boy with erythropoietic protoporphyria. AB - A 13-year-old boy who had had recurrent photosensitive skin reactions due to erythropoietic protoporphyria from 18 months of age, suddenly developed rapidly progressive hepatic failure with increasing cholestatic jaundice and variceal bleeding. Liver biopsy confirmed extensive protoporphyrin deposition with cirrhosis, and so orthotopic liver transplantation was performed. Postoperatively his skin rash settled within 72 hr, and in spite of subsequent exposure to the sun he has had no further skin reaction or blistering, although he does still have some itching. He made a good recovery and was able to return to school within six months of operation. Prior to liver transplantation, the hepatic ferrochelatase activity was reduced to only 0.81 nmol zinc-protoporphyrin formed/mg protein/hr (controls 3.30 +/- 1.00 nmol zinc-protoporphyrin formed/mg protein/hr, while the red cell protoporphyrin level was markedly elevated at 188 mumol/L red cells (normal less than 1.6 mumol/L red cells). The free plasma porphyrin level of 0.95 mumol/L (normal less than 0.02 mumol/L), and the urinary and fecal porphyrin levels were also raised. Following liver grafting these elevated porphyrin levels fell rapidly, with the red cell protoporphyrin level dropping to 10% of its preoperative value, and the rest returning to virtually normal within three months of operation. PMID- 3047930 TI - High incidence of monoclonal immunoglobulins in patients after liver or heart transplantation. AB - Sera from 56 recipients of liver or heart transplants were investigated for monoclonal immunoglobulins (mIg) by immunofixation electrophoresis (IFE) at different times during 4 years after transplantation. Transient, changing, or stable mIgs were found in 18 patients. A significantly increased mIg incidence was observed in heart Tx patients, patients over 40 years of age, and those receiving azathioprine or antithymocyte globulin in addition to prednisolone and cyclosporine as immunosuppressive treatment. No correlations could be found between the presence of mIg and the number of rejection episodes or intercurrent infections. Such serum mIg represent monoclones of at least 1 x 10(9) cells of B lymphocyte lineage that apparently proliferate without adequate suppressive control. Since immunosuppressed allograft recipients are at risk of developing B cell lymphomas, such monoclones may be regarded as prelymphomas necessitating a careful follow-up in these patients. PMID- 3047927 TI - Histocompatibility and liver transplant outcome. Does HLA exert a dualistic effect? AB - An analysis of more than 500 liver transplants has demonstrated that HLA compatibility is associated with diminished allograft survival. Liver transplants with zero mismatches for class I and/or class II HLA antigens have shown significantly lower actuarial survival rates than transplants with one or more mismatches for these loci. In a group of 119 failed liver allografts from patients undergoing retransplantation, a higher incidence of failure due to rejection correlated with a lower degree of HLA compatibility especially for HLA DR. In contrast, the incidence of liver transplant failures due to primary nonfunction was relatively higher with HLA-DR compatible transplants. Considering the role of HLA as a restriction element in cellular interactions during the immune response, these findings suggest that HLA compatibility may have a dualistic effect on liver transplant outcome. On one hand, HLA compatibility reduced transplant rejection--and on the other hand, it may enhance other immunological mechanisms leading to allograft dysfunction, particularly in patients at risk of developing recurrent autoimmune diseases or infection. PMID- 3047931 TI - Blood cyclosporine pharmacokinetics in patients undergoing marrow transplantation. Influence of age, obesity, and hematocrit. AB - Blood cyclosporine pharmacokinetics were studied in 85 patients aged 1-52 (median: 22) years undergoing allogeneic bone marrow transplantation for the treatment of hematologic disease. Pharmacokinetic studies were carried out during the first two weeks posttransplant after an intravenous dose of 2.1-4.4 mg/kg. Whole-blood cyclosporine concentrations were measured by high-performance liquid chromatography. Multiple stepwise regression analysis indicated that age (P less than 0.001) and hematocrit (P less than 0.05) correlated with cyclosporine clearance (CL) while steady-state volume of distribution (Vss) did not correlate with any of the factors studied. Cyclosporine CL significantly differed among nonobese patients in different age groups; patients less than or equal to 10 years old had a higher mean CL (13.1 ml/min/kg) than patients 11-20, 21-30, 31 40, or greater than 40 years old (mean CL: 8.5-10.3 ml/min/kg) (P less than 0.05). No significant differences in cyclosporine CL and Vss were observed between obese (greater than 125% ideal body weight) patients and age-matched nonobese patients. Hematocrit values (range: 24-39) were inversely correlated with cyclosporine CL, which suggests that red blood cells function as important ligands in cyclosporine binding. These results show that blood cyclosporine CL is higher in marrow transplant recipients than in solid organ transplant recipients and that these differences may be related to lower hematocrits in marrow transplant recipients compared with solid organ transplant recipients. When compared with previously published serum cyclosporine CL data, our findings suggest that age-related changes in CL are primarily related to changes in plasma protein binding and that obesity does not significantly alter cyclosporine CL and Vss. PMID- 3047932 TI - Changed composition of high-density lipoprotein subclasses HDL2 and HDL3 after renal transplantation. AB - The concentrations of cholesterol in the high density lipoprotein (HDL) fraction and the HDL2 and HDL3 subclasses were compared in 333 male renal transplant recipients, 36 male patients on maintenance hemodialysis, and 43 healthy men. The subclasses were separated by a precipitation method using polyethylene glycol 6000 and dextran sulphate. In hemodialyzed patients, total HDL cholesterol and both subclasses were reduced. In renal transplant recipients, both total HDL cholesterol and HDL2 were normal, whereas HDL3 remained reduced, analogous to hemodialyzed patients. It can be concluded that a successful renal transplantation has a beneficial effect on HDL metabolism and thus on the development of atherosclerosis. PMID- 3047933 TI - Genetic aspects of the blood transfusion effect. AB - The genetic requirements for the induction of the blood transfusion effect have been investigated in a genetically well-defined model. The survival of fully vascularized, heterotopic murine cardiac allografts was measured after a single preoperative transfusion of blood from mice selected to share MHC and/or minor histocompatibility (miH) antigens with the organ donor. The effect of a transfusion from a donor unrelated to the heart donor (third-party transfusion) on allograft survival was also determined. Five strain combinations were used for these experiments. The results obtained illustrate a number of important aspects of the blood transfusion effect in this model: (1) Donor-specific blood transfusion, where MHC and miH were shared by the blood donor and the organ donor, always induced prolonged graft survival. (2) The sharing of the whole MHC (H-2) by the blood donor and organ donor was found to be sufficient to prolong allograft survival in the five fully allogeneic strain combinations tested. (3) The sharing of miH antigens only was not sufficient to induce prolonged cardiac allograft survival. Special cases were identified showing that several factors could interact to potentiate the action of miH antigens in the induction of the blood transfusion effect. (4) Transfusion with blood from a third-party donor was effective in some strain combinations. In one recipient, blood from several third party strains of mice, sharing neither MHC nor miH antigens with the organ donor, induced prolonged graft survival. We suggest that the mechanism by which third party blood has a beneficial effect on graft survival is through crossreaction(s) between the blood donor and the organ donor. The results obtained in this study fit very well with one model for the cellular mechanism by which the transfusion effect may be mediated. This may be the means by which blood from randomly selected donors has a beneficial effect on graft survival in clinical transplantation. PMID- 3047934 TI - The effect of cyclosporine on the induction of unresponsiveness in antilymphocyte serum-treated, marrow-injected mice. PMID- 3047935 TI - Dependency of cyclophosphamide-induced skin allograft tolerance on age of adult recipient mice. PMID- 3047936 TI - A new, simple method for cold storage of the pancreas using perfluorochemical. PMID- 3047937 TI - Stapled duodenal-cystic anastomosis in pancreas transplantation. PMID- 3047938 TI - Hairy leukoplakia-like lesions in immunosuppressed patients following bone marrow transplantation. PMID- 3047939 TI - When further ALG administration becomes futile. PMID- 3047940 TI - Cyclosporine and its metabolites in mother and baby. PMID- 3047942 TI - Successful cardiac transplantation across an ABO blood group barrier. PMID- 3047941 TI - Mycotic aneurysm of the hepatic artery complicating human liver transplantation. PMID- 3047943 TI - Specific suppression of human liver recipients' blastogenic response to donor cells by autologous serum. PMID- 3047944 TI - Posttransplantation T cell lymphoma of the skin. PMID- 3047945 TI - Acetazolamide and cyclosporine. PMID- 3047946 TI - Pathophysiology and treatment of acute pancreatitis. PMID- 3047947 TI - Patients with surgical jaundice should always have cholangiography before operation. PMID- 3047948 TI - Cellular junctions in a spectrum of human malignant tumors. AB - Cellular junctions in tumors are often considered a hallmark of epithelial differentiation. However, junctions are also seen in tumors having a different differentiation. This observation prompted us to study cellular junctions in malignant nonepithelial tumors. We found a variety of cellular junctions in such tumors, although the majority were poorly formed. This observation is of importance for diagnostic purposes. We have also tried to clarify the nomenclature of cellular junctions as applied in tumor diagnosis by proposing a systematic categorization of terms in everyday use by pathologists and by referring more extensively to the term paired subplasmalemmal densities (PSD) for non-well-formed junctions. PMID- 3047949 TI - Incidence and ultrastructure of rudimentary cilia in benign and malignant peripheral nerve tumors. AB - The incidence and ultrastructure of cilia were examined in 26 peripheral nerve tumors. The cilia had the common features of elongated shafts and basal structures consisting of two basal bodies. The basal body contained two basal feet and some long striated rootlets. The cilia lacked central tubules and two dynein arms of subfiber A, suggesting that they were nonmotile and rudimentary. The quantitative results showed that the incidence of cilia was low in Schwann cells and fibroblasts of the 20 benign tumors and slightly higher in perineurial cells of the 12 neurofibromas. Conversely, the incidence was high in most of the 6 malignant tumors. Only one perineurial cell from normal mature nerve examined carried a cilium, although no cilia were seen in the other perineurial cells, Schwann cells, and fibroblasts observed. The results showed that ciliary formation was more marked in the more actively proliferating tissues such as malignant tumors, and less so in the less actively proliferating tissues such as benign tumors. Ciliary formation may be one of the ultrastructural features commonly observed in proliferative tissues. PMID- 3047950 TI - J.E. Purkyne memorial lecture. PMID- 3047951 TI - [Staghorn calculi. Epidemiology, pathogenesis, symptoms and treatment]. PMID- 3047952 TI - [Intestinal dialysis]. PMID- 3047953 TI - [Bronchoscintigraphy in research on the effect of terbutaline on mucociliary clearance in health individuals]. PMID- 3047954 TI - [Postoperative radiotherapy in cancer of the rectum and rectosigmoid. A prospective randomized multicenter study]. PMID- 3047955 TI - Penetrating head injuries. PMID- 3047957 TI - Recurrent urinary tract infection in the female patient. PMID- 3047958 TI - Behavioral aspects of urinary tract infection. PMID- 3047956 TI - Renal replacement treatment for diabetic nephropathy in Northern Ireland 1979 1987. AB - Twenty-three patients with end-stage renal failure due to diabetic nephropathy received renal replacement treatment. All patients had insulin-dependent diabetes mellitus. Nineteen transplants were performed in seventeen patients. Two-year graft survival for all transplants was 74% with a two-year patient survival post transplantation of 81%. Overall two-year patient survival was 73%, compared with 82% in non-diabetic patients receiving renal replacement treatment. In diabetic patients accepted for treatment there was a high incidence of non-renal complications, particularly vascular disease. An aggressive approach to the treatment of vascular disease in these patients may improve overall survival rates. PMID- 3047959 TI - Urinary tract infections in the male patient. PMID- 3047960 TI - Nosocomial infection: risks associated with short-term and long-term inpatient care. PMID- 3047961 TI - Urology update: focus on UTI. Structure--introduction. PMID- 3047962 TI - [A new method of correcting pathological blood flow in the crural veins]. PMID- 3047963 TI - [Closed injuries of the chest]. PMID- 3047964 TI - [Zollinger-Ellison syndrome]. PMID- 3047965 TI - [Giant echinococcosis of the liver simulating liver cirrhosis with pressure ascites]. PMID- 3047966 TI - [Dynamics of indicators of dermocytograms and reparative capacity of the body in burns]. AB - The investigation of dermocytograms by a modified method of the "skin window" in 101 patients with different square surfaces of deep burns has shown that the dynamics of mononuclear content is dependent on the injury degree. Lower parameters of mononuclear content in dermocytograms during the course of treatment correspond to the disturbance of reparative capacities of the organism in groups of patients with the complicated course of the disease. The "skin window" test can be used for the treatment since it shows the degree of the burn trauma dynamics and character of its course, facilitates the prognosis of complications. PMID- 3047968 TI - Highlights of contemporary research on host immune responses to ticks. AB - Host immune responses to ticks have been known since the early part of this century. This research has emanated from throughout the world with the most detailed studies originating from Australia and the United States. A review of all the studies to date indicates a diverse tick species list consisting primarily of Ixodid ticks, but a few Argasid species have been examined. Typically, research on this topic during the first half of this century has utilized the bovine host, whereas research over the past 20 years has concentrated on the rodent host. The emphasis of this research has been to define host resistance in terms of behavioral and physiological changes in the host, accompanied by changes in the feeding and reproductive potential of the ticks. The primary objective of this research is to develop an innovative tick control strategy that will allow greater and safer control than that afforded by acaricides. This paper highlights the study of host immune responses to ticks over the century to date. However, owing to the great growth in the fields of immunology and molecular biology, the greatest gains have been made from 1970 to 1985. Therefore, the emphasis of this review is on research reported during the last 15 years. PMID- 3047967 TI - [Effect of heparin on the pulmonary surfactant system in experimental studies]. AB - Experiments were performed in 9 dogs and 20 rats. The effect of heparin (3-4 mg/kg of the body weight) on the lung surfactant was studied under conditions of bacterial endotoxic shock. Heparin was found to decrease the surfactant content in lungs by 57.6% in intact animals, while under conditions of endotoxic shock by 92.2% as compared with the initial level. PMID- 3047969 TI - [Hypoinsulinemia and bioenergetic processes in the rat cerebral cortex]. AB - Increase in activity of lactate dehydrogenase, of its isoenzymes LDH1, LDH2 and LDH3 as well as in content of lactic acid with simultaneous decrease in the pyruvic acid content were found in brain hemispheres of rats with alloxan diabetes. A decrease in the rate of energy reactions in rat brain hemispheres, caused by impairment of glucose oxidation in the Krebs cycle, proved to be the specific property of metabolic alterations in these cells under conditions of hypoinsulinemia. PMID- 3047970 TI - [Iron metabolism and metalloproteins (review)]. AB - Problems of ferrokinetics, participation of metalloproteins transferrin, ferritin and lactoferrin in metabolism of iron at the step of the metal absorption, transport of iron by means of transferrin, haptoglobin and hemopexin, interaction of transferrin with reticulocytes, deposition of iron in ferritin, mobilization of iron from ferritin via ceruloplasmin are considered. Importance of blood serum ferritin is discussed. PMID- 3047972 TI - [The role of inhibitors of the complement activation system in the pathogenesis and therapy of insulin-dependent diabetes mellitus (review)]. PMID- 3047971 TI - [The effect of altitude on insulin binding by erythrocytes in experimental alloxan diabetes]. AB - Alterations in content of glucose, insulin and glucagon in blood as well as binding of insulin with specific erythrocyte receptors were studied in 120 rats with alloxan diabetes and in intact animals. Content of insulin and glucagon was decreased in rat blood within 14 days of maintaining at high altitude, whereas glycemia developed only slightly. Binding of insulin with erythrocytes was increased more distinctly in control rats, which exhibited low index of this function at low altitude. Importance of this mechanism in activation of cellular glycolysis, an increase of 2,3-diphosphate glucose level in erythrocytes as a compensatory response to high altitude hypoxia are discussed. PMID- 3047973 TI - [Conservative surgery in the early stages of breast cancer]. PMID- 3047974 TI - [Selection of the optimum method in the treatment of patients with stage Ib cervical cancer]. PMID- 3047975 TI - [Analysis of dissertations on antineoplastic chemotherapy presented 1981-1985]. PMID- 3047976 TI - [Clinico-epidemiological significance of environmental carcinogenic factors in the emergence of a hereditary disposition to tumor growth]. PMID- 3047977 TI - [Nuclear magnetic resonance in experimental oncology]. PMID- 3047978 TI - [Somatomedin activity of the blood: hormonal metabolic and immunological shifts in cancer patients]. AB - Blood somatomedin activity observed in patients with cancer of the corpus uteri, breast, colon and rectum, pancreas, ovary and lung proved to exceed those in control by 140-300%. The said parameter was found to depend upon endocrine metabolic changes, relatively higher levels being registered in overweight patients and those with hypertriglyceridemia. Changes of varying degree in somatomedin activity and growth hormone levels produced by insulin and glucose in patients with breast and uterine cancer pointed to hormonal dependence of the parameter. Also, the effect is a circumstantial evidence for negative feedback between growth hormone and somatomedin production. In patients with cancer of the breast, uterus and lung, a significant correlation was established between somatomedin activity, on the one hand, and monocyte. B-lymphocyte and T-active lymphocyte levels, on the other, matched by an inverse one with T-lymphocyte level and blastogenic reaction of lymphocytes. PMID- 3047979 TI - A fluorescent microsphere method for the investigation of erythrocyte chimaerism after allogeneic bone marrow transplantation using antigenic differences. AB - A method for the investigation of erythrocyte chimaerism in patients after bone marrow transplantation (BMT) has been developed using fluorescent microspheres coated with anti-human IgG. Blood group antigens different in patient and donor were used as marker. The specificity and reliability of this method was evaluated measuring artificial mixtures of blood group positive and negative red cells. Red cell antigens tested were D, c, K, Fya, Fyb, Jka, Jkb, S and s. Mean linear regression lines for mixtures with percentages positive cells ranging from 0.01 to 1% and 1 to 100% were 0.91X-0.03 (r = 0.99) and 1.00X-0.05 (r = 0.99), respectively. The sensitivity level of the assay was one positive cell per 10,000 blood group negative cells. In negative control samples (n = 15) a mean percentage positive cells was found of 0.002 +/- 0.004 (SD). Intra- and inter assay coefficients of variation were 4.9 and 5.3% for mixtures of 10%, and were 11.1 and 24.6% for mixtures of 0.1% blood group positive cells. It is concluded that the described method can be used both to establish the take of donor marrow in an early phase after transplantation and to investigate mixed erythrocyte chimaerism long after BMT. PMID- 3047980 TI - Cryoprecipitate for the treatment of classic hemophilia: a contribution of Judith Pool. PMID- 3047982 TI - [Development of medical microbiology in the Ukraine]. PMID- 3047981 TI - [Escherichia infection in humans and animals in the Ukrainian SSR]. PMID- 3047983 TI - [Founding and development of Lvov scientific medical schools]. PMID- 3047984 TI - [Treatment of cardiac glycoside poisoning (review of the literature)]. PMID- 3047985 TI - [Calculation of the phase structure of the cardiac cycle and central hemodynamics on the Elektronika B3-21 computer]. PMID- 3047986 TI - [Clinical pharmacology of legalon (review of the literature)]. PMID- 3047987 TI - [Functional state of the liver in patients with chronic diffuse bronchitis (review of the literature)]. PMID- 3047988 TI - [Vestibular analyzer and its role in human activity (review of the literature)]. PMID- 3047989 TI - [Lipid peroxidation in the pathogenetic and clinical aspects of infectious diseases]. PMID- 3047990 TI - [Effect of obsidan and labetalol on the course of myocardial infarction]. PMID- 3047992 TI - [Characteristics of forming international medical relations by the scientists of the Ukraine in the 18th and 19th centuries]. PMID- 3047991 TI - [Use of flugalin and profenid in patients with rheumatoid arthritis]. PMID- 3047993 TI - [The use of optimal doses of phenobarbital in the treatment of Gilbert's syndrome]. PMID- 3047994 TI - [Current data on intrahepatic cholestasis (review of the literature)]. PMID- 3047995 TI - [Fluorescent diagnosis of tumors based on using porphyrin compounds and laser irradiation (review of the literature)]. PMID- 3047996 TI - A short history of the International Association of Endocrine Surgeons. PMID- 3047997 TI - Histochemical demonstration of thyroxine, triiodothyronine, and thyroglobulin in the primary lesion of thyroid carcinoma, and its predictability for radioiodine uptake by metastatic lesions. PMID- 3047998 TI - Studies on in vivo and in vitro release of intact parathyroid hormone using a new two-site immunochemiluminometric assay. PMID- 3047999 TI - Duplex scan-derived thyroid blood flow in euthyroid and hyperthyroid patients. PMID- 3048000 TI - Benign Hurthle cell tumors of the thyroid: a diagnosis to be trusted? PMID- 3048003 TI - Nephrotoxicity: a rational approach to target cell injury in vitro in the kidney. AB - 1. The kidney is a complex organ in which there is cellular heterogeneity. Many nephrotoxic chemicals target preferentially for discrete cell types, but adjacent, morphologically different cells are unaffected. This selectivity has made the assessment of nephrotoxicity in vivo (and the study of underlying mechanisms) difficult. Discrete renal injury can, however, be exploited in vitro, to study the interactions between the toxic compound and the target cell. 2. Several in vitro models have been used to study the potential interaction between the target cells and chemicals, including perfusion of the isolated kidney, renal slices, freshly isolated fragments, primary cultures and continuous cell lines. Where appropriate, isolated organelles and purified enzymes can also be used. 3. The target cell toxicity in vivo of adriamycin, 2-bromoethanamine and hexachlorobutadiene N-acetyl cysteine conjugate is selectively maintained towards glomerular epithelial, medullary interstitial and proximal tubular cells, respectively, in vitro, showing that the "in vivo-in vitro gap" can be bridged. Characteristics unique to each of these renal cell types, such as the selective uptake of a toxin, enzyme systems for generating biologically reactive intermediates, and the presence of lipid droplets (rich in polyunsaturated fatty acid) and peroxidase activity have been identified, and one or more of these may explain the mechanisms of selective injury in discrete regions of the kidney. PMID- 3048001 TI - Familial occurrence of differentiated, nonmedullary thyroid carcinoma. PMID- 3048002 TI - Immunohistochemical study of medullary thyroid carcinoma: relationship of clinical features to prognostic factors in 36 patients. PMID- 3048005 TI - [Performance-oriented medical education in Germany 1933-1945]. PMID- 3048004 TI - Hepatotoxicity and molecular aspects of hepatocyte function in primary culture. AB - 1. The application of primary cultures of hepatocytes in testing for hepatotoxicity of drugs is reviewed. 2. Hepatotoxicity results principally from the biotransformation of toxic agents. This process is very complex and specific and involves a powerful system of multigenic isozyme families for both phase I and phase II drug metabolizing reactions. Many of the isozymes are specifically expressed in the liver in relation to the maturation or differentiation state, and are specifically induced, possibly through a complex temporally programmed gene regulation. 3. This highly specific, coordinated, molecular regulation is difficult to maintain in vitro. Isolation of hepatocytes induces a prompt differential decline of liver-specific gene transcription, which leads to preferential loss of the most specific functions, including those of the drug metabolizing isozymes, whereas repair of cell damage remains active. 4. The use of serum-free, hormonally defined media stabilizes specific hepatic functions, but not transcriptional activity, for 4-5 days. Defined media retain active DNA replication but do not permit clonal growth of hepatocytes. Co-culturing hepatocytes with primitive biliary cells prolongs cell survival and their functional capacities for several weeks, including some of the transcriptional activity. PMID- 3048006 TI - [Developmental trends in occupational medicine health protection in Germany (1870 1933)]. PMID- 3048007 TI - Demonstrating evolutionary relationships between macromolecular sequences through mutual relationships with a third sequence. AB - Corresponding sites of the Euglena chloroplast and yeast small subunit ribosomal RNAs (rRNAs) show only an insignificant match with each other but show extensive matches with Euglena chloroplast tRNA(arg). The match with the tRNA extends farther toward the 5' end of the Euglena rRNA and toward the 3' end of the yeast rRNA. The expected number of such configurations given the number of RNAs searched is about 1 in 100,000. Comparison of two sequences with a third sequence frequently reveals relationships where pairwise comparisons fail to do so. PMID- 3048008 TI - Anti-idiotypic antibodies as immunogens: idiotype-based vaccines. AB - Numerous studies have documented that antibodies may regulate the immune system and form the basis of vaccines, namely anti-idiotype vaccines. Antibodies carry individual idiotype antigenic determinants against which antibodies can be formed. When the anti-idiotype recognizes the same site that recognizes the primary antigen, a mirror image or combining site antibody may be generated. Other anti-idiotypes which recognize non-combining antigenic determinants have also been used. The evidence is reviewed for the existence of a broad range of anti-idiotypes and details are given of how an anti-idiotype vaccine based on the hepatitis B surface antigen has protected against virus challenge in the most relevant animal model system, namely the chimpanzee. Furthermore, the definition of the CD4 molecule as the conserved binding site for all known human and similar immunodeficiency viruses, (in marked contradiction to their varied neutralizing properties) has led to the raising of anti-idiotypes in mice based on the CD4 receptor which have the capacity to neutralize a broad range of isolates. PMID- 3048009 TI - Protection against virulent H5 avian influenza virus infection in chickens by an inactivated vaccine produced with recombinant vaccinia virus. AB - A cloned cDNA copy of the haemagglutinin (HA) gene of A/Chicken/Scotland/59 (H5N1) influenza virus has been expressed in vaccinia virus. This pox virus is poorly infectious or non-infectious for chickens. However, immunization of chickens with lysates of cell cultures infected with the recombinant vaccinia virus, that had been emulsified with adjuvant and which contained an estimated 0.5 microgram influenza HA, elicited a substantial neutralizing antibody response to influenza virus. Challenges of immunized and non-immunized adult chickens with virulent A/Chicken/Scotland/59 influenza virus showed that the immunized animals were highly protected while the non-immunized controls died. Immunized birds were also protected against infection with the recent virulent H5 avian influenza virus, A/Chicken/Pennsylvania/83 (H5N2). PMID- 3048010 TI - Adjuvant activity of Escherichia coli heat-labile enterotoxin and effect on the induction of oral tolerance in mice to unrelated protein antigens. AB - The ability of Escherichia coli heat-labile enterotoxin (LT) to influence the induction and maintenance of tolerance was examined in animals primed orally with a soluble protein antigen, ovalbumin (OVA), or in animals primed orally with two unrelated protein antigens administered simultaneously, OVA and bovine serum albumin (BSA). LT is immunologically and structurally related to the cholera enterotoxin (CT), which has been shown to be capable of abrogating oral tolerance to protein antigens when delivered simultaneously with the antigens. In this study, simultaneous administration of LT with OVA was shown to prevent the induction of tolerance to OVA and to increase the serum anti-OVA IgG response 30- to 90-fold over OVA-primed and PBS-primed animals, respectively. This effect was determined to be a function of the enzymatically active A subunit of the toxin since the B (binding) subunit alone was unable to influence tolerance induction. Animals fed LT with OVA after the initial OVA prime developed a significantly lower serum IgG and mucosal IgA anti-OVA response than those fed LT with OVA in the initial immunization, indicating that prior exposure to the antigen reduces the effectiveness of LT to influence tolerance and its ability to act as an adjuvant. LT was not able to abrogate tolerance once it had been established. Serum IgG and mucosal IgA responses in animals receiving LT on only a single occasion, that being upon first exposure to antigen, were equivalent to responses after three OVA/LT primes, indicating that commitment to responsiveness occurs early and upon first exposure to antigen.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3048011 TI - [Clinical and constructive aspects of extracoronal adhesive anchorage in cast prostheses]. PMID- 3048014 TI - [Autoimmune hepatitis--new aspects of histopathology, ultrastructure and immunohistology]. AB - Auto-immune hepatitis is accompanied by histological and immunohistological peculiarities which have been more thoroughly elucidated in recent time. Predominantly located in portal fields are lymphocytic infiltrations reaching continuously into peripheral lobules. Dendritic reticular cells were recordable from portal fields by electron microscopy. The hepatocytes recorded from peripheral lobules were hydropically swollen and formed microacini. Autoantibodies against components of the hepatocytic membrane seemed to be of relevance to pathogenesis and were recorded from animal experiments. Immunohistologically, the hepatocytic membrane exhibited HLA antigens and IgG. CD 8-positive lymphocytes were detected and were found to be in close contact with liver cells. PMID- 3048013 TI - Etiology of biliary cirrhosis: diagnostic features and a new classification. AB - The current classification of biliary cirrhosis, in which only a "primary" and a "secondary" form are recognized, has lost its usefulness for diagnosis, patient management, and research. In this article, a new terminology and a new etiologic classification of biliary cirrhosis and its underlying conditions are proposed, based on the current understanding of small-duct and large-duct biliary diseases and their causes. PMID- 3048015 TI - [Liver changes caused by drugs--a challenge for puncture biopsy pathology]. AB - Drug-induced liver lesions are on a steadily rising trend and raise diagnostic problem which make for a challenge to liver biopsy interpretation. History, nomenclature, and present status of this branch of hepatology are briefly discussed. The main part is devoted to problems of histological diagnosis and typing of virus hepatitis, such as drug-induced hepatitis, the clinical data necessary for judgement of these cases and the two types of drug-related idiosyncrasy so far known. PMID- 3048012 TI - [Clinical study of the new Dontisolon M]. PMID- 3048016 TI - [The nomenclature and typing of drug-induced liver changes. The opinion of a group of pathologists]. AB - A working group of histopathologists with specialised liver training has established guidelines for a nomenclature of drug-induced liver lesions. It is recommended to use the term of drug-induced hepatitis for all cases of virus hepatitis-like picture. The most frequent type is that of hepatitis with confluent necrosis. This type of hepatitis was subdivided into three subtypes of diagnostic probability for differential diagnosis versus virus hepatitis. PMID- 3048017 TI - [The future of pathological anatomy and the paradigms in science]. AB - The view is taken that the future of pathology can be perceived only in relation to the development of science in general and medical science in particular. Development of science should be seen in the dialectics of continuity and discontinuity. The power of this process depends on dialectic antagonisms, such as those between being and appearance, development of medical science and even more of biological science, and techno-scientific progress. Taking this in consideration, no principal change of trend in issues and methodology may be expected in pathological anatomy. However, differences will grow between the theoretico-experimental and clinical aspects of pathological anatomy. PMID- 3048018 TI - [25 years of the Society of Surgery of East Germany]. PMID- 3048019 TI - [Our surgical heritage. In memory of Bernard von Langenbeck, founder of natural science-oriented surgery]. PMID- 3048020 TI - Experimental treatment of Hepatozoon infections with the anticoccidial agent toltrazuril. PMID- 3048021 TI - Relation of the bovine M blood group system with growth of Streptococcus agalactiae and Staphylococcus aureus in whey. PMID- 3048022 TI - [Comparative studies of plasmid distribution in Escherichia coli (E. coli) strains from healthy and diarrheic dogs and their owners]. PMID- 3048023 TI - [Anthelmintic resistance of nematodes from cattle, sheep and goats]. PMID- 3048025 TI - Acute solitary diverticulitis of the caecum. Case report. AB - In two cases of suspected appendicitis, laparotomy revealed an inflammatory mass medially in the caecum. Colonic resection was performed in both cases and the final diagnosis was solitary caecal diverticulitis. When inflammation of a caecal diverticulum is recognized at laparotomy, simple diverticulectomy is the procedure of choice, but colonic resection is recommended if malignancy cannot be excluded or inflammatory changes are severe. PMID- 3048024 TI - Acute experimental suppurative pancreatitis in the rat. AB - Acute pancreatitis was induced in rats by infusing sodium taurodeoxycholate with or without Escherichia coli and Bacteroides fragilis into the bile-pancreatic duct. Survival did not differ between the 'noninfected bile group' (NIB) and the 'infected bile group' (IB). At standardized macroscopic evaluation, pancreatitis was more severe in the IB group (p less than 0.05). Histologic examination on day 7 showed suppuration of pancreatic tissue in 6/7 IB and 3/14 NIB rats (p less than 0.05). Bacteriologic culture of pancreatic tissue was positive in 6/8 IB and 3/17 NIB rats (p less than 0.01). Bacterial culture of blood, peritoneal fluid of pulmonary tissue was seldom positive. Concordance between microscopically observed suppuration and positive bacterial culture was almost total. Recall antigen skin testing indicated anergy in the IB group, while the NIB group showed moderately diminished reaction (p less than 0.001). Similar increase of S fibrinogen was found on day 3 in both groups, but complement factor C3 was reduced only in the IB group. This experimental model, with suppurative pancreatitis induced by intraductal infusion of an infected bile salt, may be useful for studies of systemic complications in acute pancreatitis. PMID- 3048026 TI - Gastrointestinal application of laser Doppler flowmetry. An experimental and clinical study in cat and man. AB - Laser Doppler flowmetry (LDF) was evaluated as a method for study of gastrointestinal blood flow with emphasis on clinical applicability in man. Measuring depth of LDF in the gastrointestinal tract was studied in cat and man. The LD-signal was attenuated in an exponential manner by unperfused intestinal tissue between the probe and the perfused organ. With a probe with optical fibre diameter of 0.7 mm, the measuring depth was approximately 6 mm i.e. transmural in most parts of the gastrointestinal tract. In experimental mesenteric vascular occlusion on feline small intestine, it was possible to discriminate arterial and venous occlusion by simultaneous recording of the total amount of backscattered light. The effect of epidural anaesthesia on intestinal blood flow was studied in 15 patients during large bowel resections. A significant increase of total blood flow was found in the colon (p less than 0.01) and the small intestine (p less than 0.05), with an average increase of 41% and 15% respectively. LDF was used intraoperatively for assessment of the extent and degree of disturbed microcirculation in 8 patients with late radiation injury of the gastrointestinal tract. The flow values obtained were compared with reference values earlier obtained in unaffected tissue from corresponding gastrointestinal sites. Severe reduction of blood flow was demonstrated in segments with visible fibrosis. Five segments also showed varying degrees of disturbed blood flow in adjacent macroscopically normal bowel with histological radiation injury. In patients with radiation injury of the rectum, it was possible to assess the extent of vascular damage in the rectum prior to surgery by endoscopic flowmeter recording. The post operative course was uneventful in all patients. In 23 patients with small bowel ischemia of different origins, LDF proved valuable for assessment of blood flow and tissue viability. In patients with mechanical strangulation, bowel resection was avoided in 9 out of 10 segments of clinically uncertain viability following flowmeter recording. In 6 patients with mesenteric vascular occlusion, the extent of severe ischemia seemed underestimated by clinical judgement compared to the results obtained with LDF. The ability of LDF to detect small ischemic areas and to select the proper level of resection was demonstrated. It is concluded that LDF is a suitable method for study of gastrointestinal blood flow in experimental and clinical work. LDF provides substantial information in the intraoperative management of small bowel ischemia and late radiation injury of the gastrointestinal tract. PMID- 3048027 TI - Perspectives in cytology. From Battle Creek to New Orleans. PMID- 3048028 TI - Use of monoclonal antibody HMB-45 in the cytologic diagnosis of melanoma. AB - The value of mouse monoclonal antibody (MAb) HMB-45 for the diagnosis of melanoma was retrospectively evaluated in cytologic preparations. Twenty-two (68.7%) of the 32 melanoma cases studied reacted with MAb HMB-45 while none of the 36 nonmelanoma tumors stained with this antibody. These results indicate that MAb HMB-45 can be helpful in the diagnosis of melanoma, especially in those patients who have no clinical indications of having a metastatic melanoma or in some cases in which the presence of melanin cannot be demonstrated by special histochemical stains. PMID- 3048029 TI - Cytology and immunocytology of infectious mononucleosis in fine needle aspirates of lymph nodes. AB - Two cases of infectious mononucleosis with atypical clinical presentations were initially diagnosed by fine needle aspiration (FNA) of lymph nodes and subsequently confirmed by serologic studies. The cytologic features that allowed recognition included a high percentage of cells with relatively abundant cytoplasm that stained pale to deep blue using a Giemsa-type stain. Many of the cells had plasmacytoid features. The cells ranged in size from small lymphocytes to large immunoblastic forms. Other features included mitotic figures and occasional binucleated forms. Immunologic studies showed a mixture of B and T cells, with many of the cells having a cytotoxic/suppressor phenotype. The features seem to be relatively characteristic and distinct from those of malignant processes that could be aspirated in lymph nodes. Recognition of infectious mononucleosis by FNA in these cases allowed for confirmation by serologic studies, thereby avoiding the need for an excisional biopsy. These cases show that FNA of lymph nodes may shed light on the nature of the process underlying the lymphadenopathy in selected cases of clinically atypical infectious mononucleosis. PMID- 3048030 TI - Gonadotropins. Meeting to mark the retirement of Professor Wilfrid Butt. Stratford-upon Avon. 18th September. PMID- 3048031 TI - What are we measuring in gonadotropin assays? PMID- 3048032 TI - Gonadotropins--domestic animals. AB - The application of RIAs for the gonadotropins to the study of domestic animal reproduction has depended on the availability of sensitive and specific reagents, automated systems for increased throughput and an awareness of the assay errors. Illustrations are given on the measurement of seasonal, surge and episodic changes in LH secretion and the advantages gained by such investigation are outlined. PMID- 3048033 TI - [Prognostic factors of chronic myelocytic leukemia--multivariate life table analysis of CML]. PMID- 3048034 TI - Computer analysis of circaseptanary biorhythms of sleep behaviour. PMID- 3048035 TI - The influence of hathayogic exercise Jalandhara bandha upon the cardiovascular system. PMID- 3048036 TI - J. E. Purkyne (17 December 1787--28 July 1869). PMID- 3048037 TI - The venous pattern in femoral head necrosis. Digital subtraction angiography and phlebography in 5 patients. AB - According to digital subtraction angiography in 5 cases of idiopathic ischemic necrosis of the head of the femur, the arterial and venous circulation was normal. However, pertrochanteric phlebography showed stasis of contrast indicating venous obstruction. We suggest that this could be due to arteriovenous venous shunting, possibly resulting from sympathetic dysfunction, as in the diabetic foot. PMID- 3048038 TI - Classification of congenital abnormalities of the sacrum. Patterns of associated dysfunctions. AB - Fifty-eight patients with a congenital abnormality of the sacrum were studied; a modified classification is suggested. Type 1a is characterized by an abrupt termination of the sacrum with an otherwise normal spinal column. Type 1b shows a similar sacral deficit, but with abnormalities evident higher in the spine. In Type 2 the terminal spine is malformed, and in Type 3 there is posterior sacral dysraphism. A high incidence of concomitant congenital abnormalities was found in Types 1 b and 2. Bladder dysfunction in the absence of typical clinical signs was often found in Types 1a and 1b. PMID- 3048039 TI - Large segmental necrosis of the tibia with deep infection after open fracture. AB - Fifteen patients with large segmental necrosis of the tibia with deep infection, following open fractures, were treated according to the Burri-Papineau protocol. After radical debridement, the segmental bone defects averaged 8.4 (5-15) cm. Necrectomy, external fixation, and open cancellous grafting, in a two-stage procedure, gave infection control in 14 cases and bone healing in 12. Additional procedures, i.e., plating and posterolateral bone grafting, provided union in another 2 cases. One patient underwent amputation. Functional results in the other 14, after a follow-up averaging 4 years, were good in 9 cases, fair in 3, and poor in 2. PMID- 3048040 TI - A piece of wood in the hand diagnosed by ultrasonography. AB - A piece of wood in the hand of an 11-year-old girl was not visible on radiographs, but was diagnosed by ultrasonography. Ultrasonography is useful for disclosing radiographically silent foreign bodies. PMID- 3048041 TI - [Review of the studies on the metabolites of fungi]. PMID- 3048042 TI - Studies on effects of co-culturing on the development of embryonic and postnatal cerebellar cells. AB - Experiments were conducted to compare the in vitro development of different cerebellar cells in primary surface cultures to that observed in their co cultures. Single cultures of embryonic (E.19) and postnatal (P 7) cerebellar cells as well as their co-cultures were established. Changes in cell-size distribution and in high-affinity GABA-uptake were studied during the first three weeks in the different cultures. Morphological analyses showed that in single cultures of embryonic (E 19) cells a number of large and middle-size neurons survived on the top of confluent monolayer of large flattened astroglial cells. In cultures of postnatal (P 7) cerebellar cells small tetanus toxin positive non GABAergic neurons proved to be the most abundant cell-constituents while no confluent feeder-layer of non-neuronal cells was formed. In co-cultures both embryonic and postnatal morphological features were observed but we could not observe any improvement in either survival or maturation of the neuronal types studied. A transient increase in glial GABA-uptake, however, was observed in both postnatal and co-cultures soon after plating of postnatal cerebellar cells. PMID- 3048043 TI - Plasma renin activity and electrolyte and water metabolism in normal and in genetically hypertensive (Okamoto) Wistar rats. AB - The electrolyte and water metabolisms and plasma renin activity (PRA) were investigated in genetically hypertensive (Okamoto) and in normotensive Wistar rats. The water intake, PRA, urine volume, urinary Na excretion and Na/K ratio were all found to be smaller in the genetically hypertensive rats than in the controls. No relationship could be demonstrated between PRA values and water intake. PMID- 3048044 TI - The need for computer-aided electromyography. AB - The limits of quantitative manual electromyography (EMG) are discussed. The role of computers in EMG laboratories is at present to imitate or replace the physician performing the test, extending his memory, to gather valuable information which cannot be obtained in a conventional way or, on the contrary, to delete the redundant information. The difficulties in standardizing the parameters of a single motor unit action potential (MUAP) are mainly related to the complexity of the EMG signal and its variability, particularly in pathological states. A computer-aided quantification of interference pattern is presented. The novel methods of examination applied in computerized EMG laboratories are discussed. The scope and limits of computer-aided EMG should be taken into action potential (MUAP) are mainly related to the complexicity of the EMG be accepted are listed. PMID- 3048045 TI - The diagnostic yield of automatic EMG analysis in neuromuscular diseases. AB - The aim of the study was to evaluate the diagnostic yield of automatic EMG analysis employed in differentiating normal from diseased muscle and myogenic, neural and spinal lesions. The material comprised 520 patients with neuromuscular diseases. Only diagnostically confirmed cases were included into the study. The control group comprised 51 healthy subjects. In all patients and healthy subjects routine EMG examination was performed by means of both the conventional technique and automatic method. On the basis of the statistical analysis of the material the authors concluded that the method of automated EMG using the Polish minicomputer Anops makes possible distinction of the main types of pathological processes affecting the muscles with higher than previously objectivity and reliability. They stress, however, the important role of the examiner and his experience. PMID- 3048046 TI - Methodological survey of automatic EMG analysis. AB - This paper describes a methodological survey of different methods, manual and computer-aided for quantitative MUP analysis up to the present. Due to different recording techniques and facilities for signal analysis individual laboratories have developed their own methods. Although most of the methods are still in the research phase, they provide a valuable contribution to clinical possibilities. The most important effect of the computer in electromyography is the fact that it emphasizes a new trend in quantitative EMG analysis also in routine clinical assessment. PMID- 3048047 TI - Enamine and amidine derivatives of polyene macrolide antibiotics. PMID- 3048048 TI - Past and present strategies of research on the HPA-axis in psychiatry. AB - Hypercortisolism in depression has been extensively studied during the last three decades. The main hypothesis regarding origin and clinical relevance of this phenomenon, however, has changed significantly. Up to the mid-seventies hypercortisolism was conceived as consequence of stress modified by the degree of unconscious defense mechanisms in different forms of depressive or non-depressive psychiatric disorders. At the end of the seventies this point of view changed considerably. Hypercortisolism was regarded as a biological statemarker of the endogenous subtype of depression with clinical differential-diagnostic relevance. An abnormal dexamethasone suppression test (DST) was assumed to be the best indication of increased activation of the cortisol system. These conclusions turned out to be wrong. DST results are not specific for melancholia and the test seems to be of limited value for measuring the function of the HPA-axis. Intervening variables, such as weight loss, drug and alcohol withdrawal or situational stress, influence the test results significantly, independent of the nosological classification. Additionally, interindividual differences in the susceptibility of the HPA-axis may decisively influence the the activation of the HPA-axis as well in healthy subjects under stress as in psychiatric patients. PMID- 3048050 TI - Environmental and behavioral influences on affective illness. AB - A behavioral factor, sleep, and a physical environmental factor, light, can trigger or terminate episodes of affective illness. The authors review the experimental evidence for these effects and discuss recent research on their biological mechanisms. In light of these findings the authors speculate that affective illness could partly be a disorder of systems that mediate the organism's adaptations to changes in the physical environment. PMID- 3048049 TI - Noradrenergic and serotonergic receptor system function in panic disorder and depression. AB - Recent studies which have utilized the administration of neurotransmitter agonists and antagonists to study presynaptic and postsynaptic receptor system function in panic disorder and major depression are reviewed. In panic disorder, patients have been found to be overly sensitive to the alpha-2 adrenergic agonist clonidine, and the alpha-2-adrenergic antagonist yohimbine, but they have relatively normal responses to tryptophan, the precursor of serotonin, and MCPP, a directly acting serotonin agonist. In depression, patients appear to have relatively normal responses to clonidine and yohimbine except for some supersensitivity to yohimbine effects on blood pressure and subjective symptoms. In contrast to the panic disorder patients, depressed patients demonstrate a neuroendocrine subsensitivity to tryptophan. Taken together, these studies suggest a relative abnormality in the alpha-2-adrenergic regulation of the noradrenergic system in panic disorder and a subsensitivity of the serotonergic system in depression. The pharmacologic treatment response in panic disorder and depression is similar in that both conditions respond to tricyclic antidepressants and monoamine oxidase inhibitors, but different in that panic disorder does not respond to atypical antidepressants such as trazodone and bupropion, but does respond to benzodiazepines, in contrast to depression. The possible mechanisms involved in producing these findings and the methods to more fully study possible receptor system abnormalities in these illnesses are discussed. PMID- 3048051 TI - On the contribution of sleep wake physiology to the explanation and the treatment of depression. AB - Physiological exploration has disclosed profound disturbances in the regulation of sleep in depression. The finding that depression can be relieved or intensified by manipulation of sleep has inspired the investigation of the possible pathogenetic significance of these dysregulations. Three hypotheses play a leading role in this context: the "phase-advance", the "S-deficiency" and the "acetylcholine-monoamine imbalance" hypothesis. They explain the therapeutic effects of a variety of sleep wake manipulations as consequences of the normalization of depressogenic sleep regulation disturbances. The issue discussed in this paper is whether these claims are valid. To what extent can the hypotheses explain the existing sleep physiological disturbance in depression as well as the results of therapeutic interventions in sleep? The empirical data provide firm evidence for the importance of particularly the timing of sleep in relation to the course of depression, both in respect to the spontaneous and the experimentally induced fluctuations of the clinical state. Without considerable extensions none of the hypotheses provide an adequate explanation of these facts. PMID- 3048052 TI - The Stockholm Diabetes Intervention Study (SDIS): 18 months' results. AB - Patients with insulin-dependent diabetes mellitus (IDDM), non-proliferative retinopathy and unsatisfactory blood glucose control were randomized to intensified conventional treatment (ICT, 48 patients) or regular treatment (RT, 54 patients) for a 5-year study. After 18 months the glycosylated hemoglobin (HbA1c) was reduced in both groups, but significantly more in the ICT group (p = 0.00005). Thirty of the RT patients and 16 from the ICT group deteriorated as to retinopathy (p = 0.024). Microalbuminuria appeared more often in the RT patients (p = 0.023), and nerve conduction velocities were significantly reduced only in the RT group (p between 0.0005 and 0.047). Serious hypoglycemia was more common in the ICT patients (p = 0.003). The progression of diabetic late complications was thus slowed down by intensified treatment, but at the price of an increased frequency of serious hypoglycemia. PMID- 3048053 TI - Lipoproteins and metabolic control in hypertensive type II diabetics treated with clonidine. AB - Twenty patients with type II diabetes mellitus and hypertension (WHO stages I and II) participated in a 3-month double-blind cross-over study to evaluate the effects of clonidine (75-300 micrograms daily) on blood pressure, glycemic control and plasma lipoproteins. Already after 1 month's treatment with clonidine the systolic and diastolic blood pressures had decreased, from 168/103 to 161/98 mmHg (p less than 0.01). Fasting blood glucose and HbA1c concentrations were unaffected by 3 months' treatment. Similarly, plasma lipid and lipoprotein concentrations remained unchanged throughout the study (i.e. mean high and low density lipoprotein cholesterol concentrations were 0.89 and 3.87 mmol/l on placebo vs. 0.90 and 3.98 mmol/l on clonidine). Adverse effects were mild and tolerable, and consisted mainly of dryness of the mouth. We conclude that clonidine lowers the blood pressure in patients with type II diabetes without any adverse effects on glycemic control or plasma lipoproteins. PMID- 3048055 TI - Serum neopterin and beta 2-microglobulin concentrations in monoclonal gammopathies. AB - Serum neopterin and beta 2-microglobulin concentrations were investigated in 46 patients with multiple myeloma and in 28 patients with asymptomatic monoclonal gammopathy followed for long periods (median 9.6 years) and showing an absence of evolution. Seventy-two per cent of the patients with multiple myeloma showed beta 2-microglobulin concentrations higher than 3 mg/l with a mean of 6.84 mg/l, whereas all the patients with asymptomatic monoclonal gammopathies had concentrations lower than 3 mg/l with a mean of 1.64 mg/l. Concerning serum neopterin concentrations, 91% of the patients with multiple myeloma had values with in pathological limits (greater than 8 nmol/l) with a mean of 34 nmol/l, whereas all but one of the patients with asymptomatic monoclonal gammopathy had normal values with a mean of 5.19 nmol/l. The differences thus observed in these two groups of patients are highly significant (p less than 0.001). Serum neopterin concentration, unrelated to renal insufficiency, seems to be useful in the differentiation of malignant or benign asymptomatic monoclonal gammopathies. PMID- 3048057 TI - Modifications of the single cortical vibrissal column. AB - The anatomy, cytoarchitectonics and unit responses of the vibrissal barrel field make it a good subject for studying the central effects of peripheral receptor manipulations. The paper reviews the properties of the barrel system and the evidence for columnar functioning of the barrel cortex. The plastic changes of the single vibrissal column visualized with 2-deoxyglucose autoradiography are described. Cortical plasticity was evoked by vibrissal receptor lesions, deprivation or repetitive stimulation in neonatal and adult rats. The possible processes participating in changing the appearance of the cortical column are discussed. The role of reshaping of intracortical connections within the barrel field is suggested as an important mechanism of plasticity of the vibrissal cortical columns. PMID- 3048054 TI - Inequalities in chronic dialysis and transplantation in Sweden. AB - We studied the chance of receiving dialysis and transplantation in Sweden and Stockholm based on age and sex. The number of patients accepted for chronic dialysis, divided into men and women or five age groups, were set in relation to the number of patients who died of ESRD (end-stage renal disease) in the same groups. The transplanted patients were likewise related to patients waiting on chronic dialysis. Although older patients were increasingly accepted for treatment, age is still strongly inversely correlated to acceptance. Thus in 1985, young patients aged 16-39 years had an 85% chance of being dialyzed, but patients over age 70 had only a 17% chance. Of potential male candidates, 44% received dialysis, but only 38% of women received it. Similar age, but less sex inequality, existed in the selection of patients for transplantation. In Sweden, age and sex influence the selection of patients for dialysis and transplantation. PMID- 3048058 TI - Lipoxins: a new series of eicosanoids (biosynthesis, stereochemistry, and biological activities). AB - The oxygenation of arachidonic acid and other polyunsaturated fatty acids by a wide variety of cell types results in the formation of several structurally distinct classes of biologically active compounds. These compounds include the prostaglandins, thromboxanes, leukotrienes, and other oxygenated derivatives of polyunsaturated fatty acids. A most recent addition to this family of biologically active compounds is the lipoxins (Figure 1). Leukotrienes and lipoxins are formed by mechanisms which involve initial oxygenation of free fatty acids by lipoxygenases. In general, lipoxygenase products display a wide range of actions and appear to be involved in immunity, the regulation of inflammation, and other physiological and pathophysiological processes. In this chapter we describe results of recent studies on the isolation, biosynthesis, stereochemistry and biological activities of this new series of compounds (lipoxins). PMID- 3048059 TI - Edward Neuhauser memorial lecture. Genetic bone disease: radiologic sight and insight. PMID- 3048060 TI - Wendell G. Scott memorial lecture. Breast cancer screening: all's well that ends well, or much ado about nothing? PMID- 3048061 TI - Mammographic screening in women 40-49 years old. PMID- 3048062 TI - Duplex Doppler sonography in the evaluation of adult patients before and after liver transplantation. PMID- 3048063 TI - Contribution of SPECT to imaging of gastrointestinal adenocarcinoma with 111In labeled anti-CEA monoclonal antibody. AB - Fourteen patients with suspected adenocarcinoma of the gastrointestinal tract received 1 mg of 111In-labeled anticarcinoembryonic antigen monoclonal antibody type ZCE025 combined with 40 mg unlabeled antibody of the same type. Planar and single-photon emission CT (SPECT) imaging studies were performed 3 days after infusion and, when possible, 7 days after infusion. Scan findings were correlated with the findings at surgery when possible. Tumor was detected by day-3 planar imaging in eight of 13 patients in whom tumor was documented histopathologically. Day-3 SPECT allowed demonstration of tumor in 11 of these 13 patients. In another patient whose scan was negative, no residual tumor was found at surgery. SPECT was particularly helpful in identifying small and midline tumors. In two cases, localization on SPECT helped identify the tumor mass on CT. Two primary tumors weighing less than 5 g could not be detected on either planar or SPECT scans. Histologically positive, normal-sized lymph nodes were not seen by planar imaging or SPECT. SPECT increased the detection rate over that achieved with planar imaging, helped to better localize scan abnormalities, and afforded more useful comparison between the monoclonal antibody study and CT. PMID- 3048064 TI - Diaphragmatic discontinuity associated with perihepatic ascites: a sonographic refractive artifact. AB - Apparent discontinuity in the diaphragm frequently is seen in sonograms of the right upper quadrant in patients with perihepatic ascites. Using a fresh cadaveric liver and a linear reflector to simulate the diaphragm, we performed in vitro analysis and confirmed that the appearance of the diaphragm was artifactual; this was due to refraction of the sound beam at the interface between the liver and ascites. Knowledge of the typical sonographic appearance and location of this artifact should prevent diagnostic errors. PMID- 3048065 TI - Sonography of the spermatic cord. PMID- 3048066 TI - Pitfalls in the sonographic diagnosis of uterine fibroids. PMID- 3048067 TI - Permanent Proplast temporomandibular joint implants: MR imaging of destructive complications. AB - We studied the radiologic and pathologic changes in 30 patients (34 joints) in which there were locally destructive bone and soft-tissue complications associated with previously inserted permanent temporomandibular joint (TMJ) Proplast-Teflon implants. The cases were selected as representative examples of patients with failed Proplast interpositional arthroplasty, in whom images of the TMJ were obtained with conventional radiography, tomography, and MR, and in whom both surgical and histologic findings were available. Clinical indications for imaging included joint pain, restricted joint motion, crepitus, preauricular swelling, regional lymphadenopathy, malocclusion either acquired or changed since implant surgery, and facial deformity. Surgery was then performed for the purposes of implant retrieval and joint debridement because of destructive soft tissue and osseous changes observed from the imaging analysis in conjunction with significant clinical signs and symptoms. The pathologic changes, observed 4-54 months after implant surgery, included a destructive foreign-body-type granuloma and avascular necrosis of the mandibular condyle and condylar neck. Our findings suggest that MR is useful in the detection and evaluation of destructive complications that may accompany failed Proplast-Teflon implants in the TMJ. MR is superior to conventional radiography and tomography in detecting soft-tissue lesions and avascular necrosis of bone. Tomography more accurately delineates soft-tissue calcifications and cortical margins of osseous structures. PMID- 3048068 TI - Frederic N. Silverman, 1988 Gold Medalist of the Society for Pediatric Radiology. PMID- 3048069 TI - Sonographic evaluation of spinal cord birth trauma with pathologic correlation. AB - Birth trauma to the spinal cord is a serious potential complication of delivery. Determining the presence, severity, and extent of injury poses a difficult problem because of the often confusing clinical setting. Myelography has been recommended for assessing spinal cord birth trauma but is invasive and may not be helpful. The role of sonography in evaluating spinal cord birth trauma has not been previously described. We assessed the value of sonography in four patients, three of whom also had CT metrizamide myelography. Autopsy correlation was available in three patients. Sonography was able to easily demonstrate the cord configuration, allowing for multiple assessments over time. Internal cord echogenicity was helpful in a case of hematomyelia and in demonstrating the changes of myelomalacia. Sonography is useful in evaluating neonates with severe spinal cord injury; it obviates the need for myelography and also may allow less severely injured patients to be assessed more frequently. PMID- 3048070 TI - The dangling choroid plexus: a sonographic observation of value in excluding ventriculomegaly. AB - To show that the position of the choroid plexus is dependent on gravity and to prove that this fact can be used as a simple means of avoiding the erroneous diagnosis of ventriculomegaly on fetal sonography, we evaluated 75 fetal sonograms retrospectively. Twenty-five fetuses had ventriculomegaly, and 50 had normal cerebral ventricles. The gestational ages ranged from 15 to 39 weeks. To show objectively that the position of the choroid plexus within the lateral ventricle was gravity dependent, we measured the choroid angle in each case. The choroid angle (the angle between the long axis of the choroid plexus and the linear midline echo on transverse axial sonograms through the body of the lateral ventricles) varied directly with ventricular size. In the group with normal-sized ventricles, the values for choroid angle followed a normal, unimodal distribution and had a mean of 14 degrees, a range of 6-22 degrees, and an SD of 4.3 degrees. In cases of ventriculomegaly, the values for choroid angle did not follow a normal distribution and ranged from 29 to 90 degrees. The choroid "dangled" from its attachment at the foramen of Monro and rested on the dependent wall of the lateral ventricle, resulting in a choroid angle that was increased over normal; the degree of the angle was dependent on the severity of the ventricular enlargement. The resting position of the choroid plexus marked the position of the lateral ventricular wall even when the reflection of ultrasound from the ventricular wall itself could not be seen. Detection of the position of the dependent choroid plexus is a simple observation that can be used to avoid the erroneous diagnosis of fetal ventriculomegaly and to help gauge the severity of true ventricular enlargement. PMID- 3048071 TI - MR imaging of metallic implants and materials: a compilation of the literature. AB - Ferromagnetic metallic implants and materials are regarded as contraindications for MR imaging because of the potential risks associated with their movement or displacement. To date, 14 published articles have evaluated the ferromagnetic qualities of 127 different metallic implants and other materials, including aneurysm and hemostatic clips (32); dental implants and materials (five); intravascular coils, filters, and stents (13); ear implants (14); prosthetic heart valves (29); orthopedic implants and materials (eight); penile implants (nine); and miscellaneous metallic implants and materials (17). All of these materials were evaluated by measuring the deflection forces induced by static magnetic fields at strengths ranging from 0.147 to 4.7 T. This article is a compilation of the results of these studies; it lists all 127 of the materials tested, indicates whether they were found to be deflected by the static magnetic fields, and gives the highest static magnetic field strength at which they were evaluated. Of the metallic implants tested, 66 were nonferromagnetic, and 29 exhibited only minimal deflection relative to their in vivo applications (i.e., the deflection forces were thought to be insufficient to move or dislodge the implant or material in situ). The authors of these studies concluded that patients with these particular metallic implants or materials (95/127, 75%) can be examined safely by MR imaging with scanners having static magnetic field strengths up to and including those used for the specific evaluations. Patients with other ferromagnetic materials or implants may also undergo MR imaging safely; however, both careful consideration of the factors that influence the deflection of metallic implants and prudent clinical judgment are required before patients who have these objects are examined via MR imaging. PMID- 3048072 TI - Physician perception of exercise electrocardiography as a prognostic test after acute myocardial infarction. AB - To determine how physicians interpret exercise electrocardiography with respect to prognosis after acute myocardial infarction (AMI), 29 cardiologists (all board certified) were presented a case history of a 50-year-old man with an uncomplicated AMI and asked to estimate the patient's risk of dying over the next year, the sensitivity and specificity of exercise electrocardiography with respect to 1-year mortality, and the patient's risk of dying given a positive and a negative test result. Each set of physician estimates did not differ from those derived from a review of the medical literature (difference not significant for each). Risk after the test was also calculated using the Bayes' theorem. Calculated versus estimated risks were compared after a negative (7 +/- 9 vs 11 +/- 11%) and a positive (27 +/- 22 vs 17 +/- 15%, differences not significant) test result. Estimated risks were more accurate for a negative result than for a positive one (89 +/- 10 vs 83 +/- 12%, p less than 0.001). Given a positive test result, 57% of the physicians recommended coronary angiography. However, their estimates of risk (30 +/- 23%) were not significantly different from the estimates of those physicians (14%) who recommended additional noninvasive testing (19 +/- 4%) or those (29%) who recommended medical therapy (28 +/- 26%) (difference not significant). Thus, cardiologists accurately estimated prognosis following AMI, but they were less accurate in assessing high risk than low risk, and their management decisions correlated poorly with their risk assessments. PMID- 3048073 TI - The automatic implantable cardioverter defibrillator as antiarrhythmic treatment modality of choice for survivors of cardiac arrest unrelated to acute myocardial infarction. PMID- 3048074 TI - Repeated endomyocardial biopsy causing coronary arterial-right ventricular fistula after cardiac transplantation. PMID- 3048075 TI - Reassessing the effects of simple carbohydrates on the serum triglyceride responses to fat meals. AB - The effects of glucose, sucrose, and fructose ingestion on the serum triglyceride responses to meals containing 40 g fat were studied in 21 normolipidemic, nonobese medical students (9 men, 12 women). Mean postprandial lipemia was not significantly lower after meals containing 50 g glucose and 40 g fat than after meals containing 40 g fat alone (2.11 vs 2.42 mmol.L-1.7 h-1, p greater than 0.45) despite a substantial increase in plasma insulin concentrations after the glucose-containing meal. Ingestion of 50 g fructose and 40 g fat (4.23 mmol.L-1.7 h-1, p less than 0.0001) and 100 g sucrose and 40 g fat (3.77 mmol.L-1.7 h-1, p less than 0.001) resulted in significantly greater lipemia than did the ingestion of fat alone. These findings suggest that the ingestion of insulinogenic carbohydrates does not reduce postprandial lipemia in normolipidemic subjects and that the ingestion of fructose and fructose-containing carbohydrates may augment the postprandial lipemia induced by a fat-containing meal. PMID- 3048076 TI - Relationship between the rate of gastric emptying and glucose and insulin responses to starchy foods in young healthy adults. AB - Twelve young healthy adults (five men, seven women) ingested four test meals on four occasions so we could examine the relationship between the rate of gastric emptying (GE) and the glucose response to different starchy foods. Each meal consisted of one food product containing 50 g starch: spaghetti, rice, French bread, or mashed potato. Basal and postprandial glucose and insulin responses were measured for 3 h. The foods were labeled with 3.7 MBq Tc99m-albumin and GE was studied by scintigraphy for 3 h. The rate of GE (expressed by the GE half time) was fastest for mashed potatoes, then bread, rice, and slowest for spaghetti. Blood glucose and serum insulin responses were similar. A significant negative correlation was found between the GE half-time and the maximum variation in blood glucose level (r = -0.6, p less than 0.0001). The glucose response to all four foods is strongly related to the GE rate. PMID- 3048077 TI - Glucose and insulin response in diabetic subjects: acute effect of carbohydrate level and the addition of soy polysaccharide in defined-formula diets. AB - This single-meal pilot study compared the plasma glucose and serum insulin response to defined-formula diets with two levels of carbohydrate (CHO) (55% and 30% of the kilocalories) with and without added soy polysaccharide (10 g) in subjects with type 2 diabetes mellitus. Subjects received each of the four liquid formula test meals in a randomly assigned order: 1) high CHO, low fiber (HC, LF), 2) high CHO, high fiber (HC, HF), 3) low CHO, low fiber (LC, LF), and 4) low CHO, high fiber (LC, HF). On the day of each test meal the formula was consumed, eight blood samples were drawn for plasma glucose and serum insulin measurements, and a 4-h urine collection was obtained for measuring glucose excretion. Our results showed that area increments under glucose and insulin curves were significantly lower with both low-CHO formulas (p less than 0.001). The addition of soy polysaccharide to the liquid formula did not result in statistically different area increments for glucose or insulin. PMID- 3048078 TI - On the shoulders of giants. PMID- 3048079 TI - Total body water estimated by measuring total-body electrical conductivity. AB - A second-generation total-body electrical conductivity (TOBEC) instrument for adults (HA-2) was evaluated against isotope dilution of 2H and 18O for its ability to estimate total body water (TBW) in 20 healthy adults. The highest correlation coefficient (0.997) and the lowest standard error of the estimate (0.68 kg) were obtained using the first (FC0) and third (FC2) Fourier coefficients of the transformed TOBEC signals and the variables height (m) times average lean circumference (m) and age (y) in the prediction equation of TBW as follows: TBW (H218O) in kg = 10.8 + (0.0724.FC0) - (0.221.FC2) + (0.0398.age) + (9.2.height.average lean circumference) where average lean circumference is the average of the lean chest, abdomen, and thigh circumferences. The TOBEC instrument for adults provides a suitable alternative for the estimation of TBW. PMID- 3048080 TI - Gastroduodenal complications of chronic NSAID therapy. AB - The fact that nonsteroidal anti-inflammatory drugs (NSAIDs) damage the gastroduodenal mucosa is no longer contested. Endoscopic studies in normal volunteers after NSAID administration have failed to predict which NSAIDs would be safest when administered chronically. NSAID use has been associated with a disproportionately high frequency of upper gastrointestinal bleeding and perforation of ulcers. All of the newer NSAIDs appear to be similar in their propensity to cause mucosal damage, including peptic ulceration. On any given day, more than 10% of patients receiving NSAIDs chronically will have a gastric ulcer, a point prevalence of ulcer disease at least 5 to 10 times higher than in patients who are not taking NSAIDs. The dose-response relationship between anti inflammatory activity and untoward events, coupled with increased use of newer more potent NSAIDs, explains, in part, the increased incidence of NSAID associated ulcer complication of bleeding and perforation. The possible association of the increase in prevalence of Campylobacter pylori gastritis with aging and the apparent increase in NSAID-associated complications in the elderly is discussed. The current status of nonsteroidal drug therapy can be summarized as follows: 1) new NSAIDs are not safer than the old NSAIDs, as far as major gastrointestinal side effects are concerned, 2) NSAIDs should be avoided when analgesia is the main goal, 3) if NSAIDs are required, the lowest possible dose that achieves pain relief should be used, 4) newer NSAIDs available only in relatively high anti-inflammatory activity dosages should be restricted to those patients in whom high levels of anti-inflammatory activity are desired. PMID- 3048081 TI - Immunohistochemical studies on the distribution of nerve fibers in chronic liver diseases. AB - Innervation of the liver with or without chronic liver disease was immunohistochemically studied. In normal liver tissues, gamma-enolase [neuron specific enolase (NSE)]- and S-100 protein (S-100)-positive nerve fibers were found around hepatic arteries, portal veins, and bile ductules in the portal area, along the sinusoid, and around central veins inside the hepatic lobule. Neuropeptide Y (NPY)-immunoreactive (IR) nerve fibers were detected in close contact with hepatic arteries and portal veins in the portal area and along the sinusoid in the parenchyma. Vasoactive intestinal polypeptide (VIP)-IR nerve fibers were found in the portal area but were scarce in the parenchyma. VIP-IR nerve fibers were much fewer than NPY-IR fibers. In patients with chronic active hepatitis, NSE- and S-100-positive nerve fibers, as well as NPY-IR nerve fibers, proliferated in the enlarged portal area. In cirrhotic liver, NSE- and S-100 positive nerve fibers and NPY-IR fibers increased remarkably in the fibrous septa, whereas they decreased or vanished inside the pseudolobules. These findings suggest that, in chronic liver disease, intrahepatic nerve fibers change their distribution, accompanying the structural changes in the hepatic lobules. PMID- 3048082 TI - Carcinoma of the ampulla of Vater: expression of cancer-associated antigens inversely correlated with prognosis. AB - To obtain some useful pathologic indicators for predicting the prognosis in carcinomas of the ampulla of Vater, we analyzed 24 surgically resected ampullary carcinomas pathologically with immunohistochemistry of cancer-associated antigens. Pancreatic invasion, lymph node metastasis, and histology of the tumor were significantly correlated with poor prognosis (p less than 0.01), but the size or ulceration of the tumor did not significantly affect the prognosis (p less than 0.05). Immunohistochemically, diffuse positivity for anti-CA19-9 monoclonal antibody was demonstrated in 10 carcinomas and that for anti carcinoembryonic antigen (CEA), in 10. Eight of them showed synchronously diffuse immunoreactivities for both antigens. Although there was no significant correlation between diffuse positivity for CA19-9 and pathologic factors, CA19-9 positive cases exhibited significantly poor prognoses (p less than 0.01). Diffuse positivity for CEA was correlated with pancreatic invasion (p less than 0.05) and poor prognosis (p less than 0.05). Immunohistochemical study of cancer-associated antigens may disclose some malignant potential of ampullary carcinoma other than that expressed in the morphology. Furthermore, because of the consistency of staining results, immunohistochemistry of cancer-associated antigens may also be useful in predicting preoperatively the prognosis of ampullary carcinoma in biopsied materials. PMID- 3048083 TI - Monooctanoin-associated pulmonary edema. AB - Monooctanoin is a cholesterol solvent indicated for dissolution of retained biliary stones. We summarize four reports--one from the U.S. Food and Drug Administration's Spontaneous Reporting System for Adverse Drug Reactions and three from published medical literature--of noncardiogenic pulmonary edema during intrabiliary monooctanoin in the United States. Based on these data, we show that pulmonary edema during intrabiliary monooctanoin infusion may occur in approximately one per 1000 patients treated. PMID- 3048084 TI - Choriocarcinoma of the stomach: pathogenesis and clinical characteristics. AB - A patient with primary choriocarcinoma of the stomach is reported. Abdominal pain, weight loss, a palpable epigastric mass, and gastrointestinal bleeding are the most common clinical features of this germ cell tumor, making it difficult to distinguish choriocarcinoma from primary adenocarcinoma of the stomach. The unique aspect of this case is the mixed histology of this rare neoplasm, containing both adenomatous and embryonal carcinoma, in addition to the choriocarcinoma. We believe that this unusual tumor probably results from dedifferentiation of primary adenocarcinoma of the stomach. On the average, patients with this disease live less than 2 months from the time of diagnosis, and treatment with combination chemotherapy has not improved the survival. PMID- 3048085 TI - Neoplastic angioendotheliomatosis with multifocal hemorrhagic necrosis of the liver. AB - We describe an autopsy case of neoplastic angioendotheliomatosis with an emphasis on hepatic pathology. The neoplastic cells were found mainly in small vessels and capillaries throughout the body, especially in the lung, liver, and spleen. These cells showed immunophenotypes of B-cells indicating differentiation toward B cells. The liver (1350 g) showed multiple hemorrhagic areas, especially in the subcapsular region. Microscopically, neoplastic cells were found in the dilated sinusoids, portal vein branches, and central and sublobular hepatic veins, and there was hemorrhage with hepatocytic necrosis and dropout, especially in centrilobular areas. Thromboemboli admixed with neoplastic cells were often found in portal veins and sublobular hepatic veins, which might have resulted in the multifocal hemorrhagic necrosis. PMID- 3048086 TI - An epidemiologic study of sudden death at work in an industrial county, 1979 1982. AB - The descriptive epidemiology of sudden death at work was studied in an urban, industrial county. County coroner's records were used to identify the 212 deaths that occurred at work among employed white males in Allegheny County, Pennsylvania in 1979-1982. Occupations and industries with increased risks for either sudden natural deaths or fatal injuries at work were identified by comparison with the white male county employed population. Men employed in service occupations had the highest age-adjusted sudden natural death rate at work (27.0 per 100,000). This was 2.5 times as high as the overall county rate. Men employed in the construction industry had the highest age-adjusted rate of fatal injuries at work (24.3 per 100,000). This was 4.4 times as high as the overall county rate. Twenty-five per cent (17/68) of occupational fatalities involved multiple fatalities or injuries. Only 1 per cent (2/144) of natural deaths at work and 7 per cent (5/68) of fatal injuries had blood alcohol levels exceeding 0.1 mg/100 ml, the level of intoxication. Improvements in the prevention and surveillance of sudden deaths that occur at work are suggested. Coroner's records are suggested for use in future surveillance on sudden deaths at work because they identified more sudden deaths at work than death certificates did. PMID- 3048087 TI - Investigation of an outbreak of Salmonella enteritidis gastroenteritis associated with consumption of eggs in a restaurant chain in Maryland. AB - Salmonella enteritidis ser. enteritidis was isolated from patrons and employees of three restaurants in a restaurant chain in Maryland during August and September 1985. Isolates from all three restaurants had identical plasmid profiles; this profile was present in 13 of 40 randomly selected S. enteritidis isolates received by the Maryland state health department laboratory during a comparable time period. The outbreak in one restaurant resulted in at least 71 illnesses, with 17 persons known to have been hospitalized. Scrambled eggs served on a "breakfast bar" were implicated as the vehicle of transmission in this restaurant, with eggs a possible vehicle in another of the three restaurants. The data point out the risks associated with improper handling of eggs in food service establishments, provide further evidence for the observed association between S. enteritidis and eggs in the northeastern United States, and demonstrate the utility of plasmid analysis in investigation of outbreaks involving common Salmonella serotypes. PMID- 3048088 TI - Is passive surveillance always insensitive? An evaluation of shigellosis surveillance in Oklahoma. AB - The authors studied the reporting of shigellosis in Oklahoma to evaluate the sensitivity of the state-based passive surveillance system for shigellosis. They found that passive surveillance for shigellosis can be more sensitive than has been previously observed. Laboratory-based reporting was found to be far superior to reporting by physicians. PMID- 3048089 TI - Management of urinary tract infections. AB - Community-acquired urinary tract infections account for millions of physician visits per year. When urinary tract infections develop in hospitalized patients, they not only increase the duration of hospitalization (and thus its cost) but also have a serious and sometimes devastating impact on the incidence of morbidity and mortality. Treatment depends on the clinical setting, the causative organism, the site of infection, and the patients' host defenses. Diagnosis and subsequent appropriate classification provide important information on which to base therapeutic decisions. Simple first infections, particularly in women, generally respond to treatment with a simple antibiotic drug. Complicated or recurrent infections, in contrast, require other therapeutic strategies. PMID- 3048090 TI - A double-blind, multicenter, comparative study of the safety and efficacy of cefixime versus amoxicillin in the treatment of acute urinary tract infections in adult patients. AB - In this 31-site multicenter trial, 565 adult patients with urinary tract infections were randomly assigned to receive either a 10-day course of cefixime 400 mg once daily (n = 279) or amoxicillin 250 mg three times daily (n = 286). Although all patients were included in the safety analysis, only 93 (33 percent) cefixime-treated and 99 (35 percent) amoxicillin-treated patients were fully evaluable for the efficacy analysis. One week after therapy, the evaluable patients treated with cefixime demonstrated a 90 percent clinical cure rate and a 92 percent eradication rate of the baseline pathogen. This compared with an 83 percent clinical cure rate and an 84 percent bacterial eradication rate in the amoxicillin-treated group. The most frequently isolated pathogen was Escherichia coli (80 percent) followed by Proteus mirabilis (10 percent). One hundred thirty seven (49 percent) of the 279 cefixime-treated and 126 (44 percent) of the 286 amoxicillin-treated patients reported at least one adverse experience during the study. Adverse reactions associated with cefixime were similar to those reported for other beta-lactam antibiotics. The most frequent adverse experiences reported by cefixime-treated patients were diarrhea (15 percent) and stool changes (12 percent). Headaches (11 percent) and diarrhea (9 percent) were the most frequently reported adverse reactions by the amoxicillin-treated patients. Eleven cefixime-treated patients (3.9 percent) and 10 amoxicillin-treated patients (3.5 percent) discontinued therapy because of adverse experiences. Results of this study demonstrate that a once-daily regimen of cefixime is as safe and effective as a three-times-daily regimen of amoxicillin in the treatment of acute urinary tract infections. Although the incidence of bowel changes was somewhat higher in the cefixime treatment group, these events usually resolved when therapy was discontinued. PMID- 3048091 TI - Management of acute and chronic respiratory tract infections. AB - Pharyngitis, bronchitis, and pneumonia represent the most common respiratory tract infections. With a view to establishing effective management strategies, the origins of these illnesses and the diagnostic techniques that have been developed to discover them are reviewed. Therapeutic regimens with documented efficacy are outlined with emphasis on specific rather than empiric treatment. Although many respiratory tract pathogens remain exquisitely sensitive to penicillin, the emergence of resistant strains underscores the need for safe and effective alternative therapies. PMID- 3048092 TI - Comparative, multicenter studies of cefixime and amoxicillin in the treatment of respiratory tract infections. AB - A total of 560 patients were treated in two double-blind, randomized multicenter studies to compare the safety and efficacy of cefixime (400 mg administered once daily) and amoxicillin (250 or 500 mg administered three times daily) for the treatment of bacterial respiratory tract infections. Eighty percent of the 244 patients treated in the lower respiratory tract infections (LRTI) study had acute bronchitis. Streptococcus pneumoniae (13 percent), Haemophilus influenzae (28 percent), and Escherichia coli (10 percent) were the pathogens most frequently isolated from sputum in these patients. Among evaluable patients with positive bacterial culture results at baseline, a favorable clinical response (cured or improved) was obtained in 100 percent of the cefixime-treated patients (22 of 22) and in 96 percent of the amoxicillin-treated patients (23 of 24). Bacteriologic eradication rates were 100 percent and 83 percent for cefixime and amoxicillin, respectively. In the upper respiratory tract infections (URTI) study, 316 patients with pharyngitis (80 percent) or tonsillitis (14 percent) were treated. Group A, beta-hemolytic Streptococcus (69 percent) and H. influenzae (8 percent) were the pathogens most frequently isolated from the throat culture specimens of these patients. Favorable clinical results were obtained in 99 percent of the evaluable cefixime-treated group (n = 73) and in 98 percent of the amoxicillin treated group (n = 66). The bacteriologic eradication rates were 93 percent and 100 percent, respectively. The adverse experiences reported during both studies were similar in nature and frequency to those reported for other beta-lactam antibiotics with the exception of a higher incidence of altered bowel movement (diarrhea and stool changes) with both drugs. These episodes usually resolved without remedial medication when the treatment was withdrawn. No significant adverse laboratory findings were observed. Results of these trials demonstrate that cefixime at a dosage of 400 mg once daily is an effective and safe oral antibiotic for the treatment of acute respiratory tract infections. PMID- 3048094 TI - Epidemiology of hypertension in older patients. AB - There are few studies devoted specifically to the epidemiology of older hypertensive patients, although some information has been obtained from broader trials in which either the study populations have become older or an older subgroup has been identified. Three areas have been addressed: the changes of blood pressure with age; the prevalence of hypertension in older persons; and the risks of elevated blood pressure in elderly patients. It has been found that blood pressure, especially systolic pressure, increases with age. Consequently, hypertension is extremely prevalent in this age group, affecting 65 percent or more of those over 65 years old, and the phenomenon of isolated systolic hypertension is common. Whereas in the past many physicians treating the elderly have regarded hypertension in these patients as normal and acceptable, or even helpful in ensuring adequate organ perfusion, elevated blood pressure in the elderly is far from benign. Older hypertensive patients' risks of cardiovascular complications and death are from two to five times that of normotensive persons. Treatment of diastolic hypertension in older patients can be expected to delay the onset of cardiovascular disease and improve the quality of life. Hopefully, this will prove to be true for isolated systolic hypertension as well. PMID- 3048093 TI - The evolution of current hypertension therapy. AB - There has been a continuous evolution in hypertensive therapy during the last 30 years. Now, physicians have access to more than 40 agents for treating this widespread condition. Large-scale clinical trials have established that lowering blood pressure in patients with mild to moderate diastolic hypertension results in a decreased incidence of stroke and, to a lesser extent, a reduction in incidence of coronary heart disease [MacMahon SW, Cutler JA, Furberg CD, et al: Prog Cardiovasc Dis 1986; 29 (suppl 1): 99-118]. Even so, the decrease in overall mortality rate is not consistent. Although hypertension occurs with increasing frequency in those over 60 years of age, patients in this age group represent less than 12 percent of the subjects in large trials. Currently, stepped-care is the recommended approach for managing hypertension in patients of all ages. However, the availability of a variety of agents for initial therapy, all with approximately equal efficacy but differing side-effect profiles, calls such an approach into question. PMID- 3048095 TI - Pathophysiology of hypertension in older patients. AB - More than half of the United States population over 65 years of age has essential hypertension. In 1984, there were 10 million elderly hypertensive persons and this number will reach 25 million in the near future. These patients are at high risk for congestive heart failure, stroke, heart attack, and dissecting aneurysm. Successful reduction of blood pressure can lower these risks considerably, but rational treatment depends on understanding the complex pathophysiology of hypertension in older patients. In fact, treatment that does not take into account the combined effects of aging and hypertension on the cardiovascular system and the kidneys may do more harm than the hypertension itself. Among the prominent age-related cardiovascular changes are stiffening of the arterial tree, with or without a contribution from atherosclerosis. This reduces arterial compliance and increases afterload, resulting in the left-ventricular hypertrophy seen in old age and leading to a progressive rise in systolic pressure. There is considerable shrinkage of the kidneys, due primarily to loss of glomerular and tubular tissue in the cortex, along with sclerosis of the glomeruli and formation of tubular diverticula. Arteriolar changes lead to reduced renal blood flow, the shunting of blood around the glomeruli, and thus a reduction in glomerular filtration rate. Renal water and electrolyte excretion are changed, making homeostasis more difficult to maintain, and the renin-angiotensin system is altered, helping to blunt the kidneys' response to pressure changes. Essential hypertension superimposed on all the foregoing effects exacerbates them. Peripheral resistance is usually markedly elevated in older hypertensive persons, which increases afterload directly.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3048096 TI - Vaginal ultrasound for diagnosis of placenta previa. AB - Transvaginal sonography was compared with transabdominal sonography in 35 women with suspected placenta previa. The transvaginal sonographic technique did not result in vaginal bleeding in any of the patients. The internal os and its relationship to the location of the placenta were visualized by transvaginal sonography in all patients, but only in 24 patients (69%) by transabdominal sonography. Transvaginal sonography ruled out placenta previa in 13 cases thought to be placenta previa by abdominal sonography. The transvaginal diagnosis in these 13 patients was confirmed at delivery. Thirty-four of the 35 women have been delivered. The diagnosis at delivery confirmed the transvaginal sonographic diagnosis in 29/34 cases and the transabdominal diagnosis in 16/34. Transvaginal sonography did not predict the delivery diagnosis in five patients who were erroneously believed to have placenta previa by both sonographic techniques. PMID- 3048097 TI - Evaluation of ultrasound diagnosis of fetal anomalies in women with pregestational diabetes: University of Florida experience. AB - Fetal congenital anomalies have become the major cause of perinatal morbidity and mortality in pregnancies complicated by insulin-dependent diabetes. We evaluated the use of level II ultrasound in predicting congenital anomalies, to determine if the management of pregnant women with insulin-dependent diabetes would be altered by these findings. We examined 43 insulin-dependent diabetic pregnancies. In this group, 10 newborns (23%) were diagnosed either at birth or later to have an abnormality. Of these, four (9%) were diagnosed by ultrasound. Of the seven cases that were undiagnosed, three women did not undergo the level II examination, two women had lesions undetectable by ultrasound, and in two women cardiac abnormalities were missed. Of the four congenital anomalies that were prenatally diagnosed, the findings influenced the management in three cases. The level II ultrasound used as a screening test had a 67% sensitivity and a 100% specificity, with a positive predictive value of 100% and a negative predictive value of 91%. We conclude that high-resolution ultrasound may be used as a screening tool for congenital anomalies in the insulin-dependent diabetic pregnancy. This study also suggests that the use of fetal echocardiography with evaluation of aortic and pulmonary outflow tracts, the arch of the aorta, and ventricular size may be helpful in the diagnosis of fetal cardiac anomalies in the pregnant woman with insulin-dependent diabetes. PMID- 3048098 TI - Gestational diabetes reverses the circadian variation of plasma insulin response to intravenous glucose. AB - Both healthy third-trimester pregnant women and a group of women with gestational diabetes failed to show a difference in glucose clearance rates when given an intravenous glucose bolus at 8 AM compared with 4 PM. The plasma insulin response in the healthy pregnant women was greater at 8 AM. In the diabetic group, the peak insulin response was greater at 4 PM, but it was more prolonged after the 8 AM tests. These alterations in plasma insulin response were especially striking in the subgroup of obese women with gestational diabetes, who demonstrated metabolic differences compared with their nonobese counterparts. PMID- 3048100 TI - Efficacy of the fetal-pelvic index in patients requiring labor induction. AB - The fetal-pelvic index is a recently described means of identifying the presence or absence of fetal-pelvic disproportion. In this follow-up study, the efficacy of the fetal-pelvic index was evaluated in 49 patients requiring labor induction and was compared with two other methods used to identify fetal-pelvic disproportion (Colcher-Sussman x-ray pelvimetry and ultrasound-derived estimated fetal weight). Twelve of 14 patients who required operative intervention also demonstrated a positive fetal-pelvic index (sensitivity = 86%). Spontaneous vaginal deliveries occurred in 35 patients, all of whom demonstrated negative fetal-pelvic index values (specificity = 100%). Overall the predictability of the fetal-pelvic index was 96%. In comparison, when used alone, neither x-ray pelvimetry nor ultrasound-derived estimated fetal weight accurately detected the presence or absence of fetal-pelvic disproportion in patients requiring labor induction. PMID- 3048099 TI - Pregestational diabetes: insulin requirements throughout pregnancy. AB - The management of pregestational diabetes requires tight metabolic control to reduce maternal and perinatal morbidity and mortality. It has been suggested that type I diabetes is a disorder characterized by insulin deficiency and type II diabetes is characterized by insulin resistance; however, it may be hypothesized that a difference in insulin requirements should emerge throughout pregnancy to reflect the dissimilarities in these two metabolic disturbances. The current investigation of 103 women with pregestational diabetes used a novel approach (reflectance meters with onboard memories) to uncover the actual insulin dosages required to reach and maintain optimum metabolic control throughout pregnancy. It was found that both type I and type II diabetes appear to have a triphasic insulin pattern, with the patient having type II diabetes requiring significantly higher doses of insulin during each trimester. This seems to suggest that the hormonal changes in pregnancy may have a similar effect on both type I and type II diabetes but to a different degree. Thus this should be considered in the treatment of pregestational diabetes and in the development of an algorithm for diabetes management. PMID- 3048101 TI - A close look at early embryonic development with the high-frequency transvaginal transducer. AB - Transabdominal sonography has been, for the past two decades, used as an effective diagnostic and research tool in obstetrics. It is predominantly used in the second and third trimesters of gestation. Its use in the first trimester is relatively limited and mostly diagnostic in nature. The introduction of the higher frequency transvaginal transducer probe, with its higher resolution of the images, opens new possibilities to study early gestation. We studied embryonic development in 38 well-dated and normal pregnancies. A well-defined intrauterine gestational sac could be seen at 4 weeks and 1 to 4 days of menstrual age. The beta-subunit of human chorionic gonadotropin level at this time was 450 to 750 mlU/ml. Structures such as the yolk sac, membranes, ventricular system in the brain, musculoskeletal system, and cord were described and illustrated. Textbooks and atlases were used for comparative purposes. High-resolution transvaginal sonography will facilitate first-trimester perinatology. PMID- 3048103 TI - Efficacy and side effects of magnesium sulfate and ritodrine as tocolytic agents. AB - Ritodrine as the first-line drug in the treatment of established preterm labor has been supplanted in some centers by magnesium sulfate. To assess the relative efficacy and rates of side effects of these two agents, 120 patients were randomly assigned to receive one of these two drugs. Patients were included if they had intact membranes and met strict criteria for the definition of labor. In both groups excellent outcome was achieved, with 96.3% and 92.3% of patients receiving ritodrine and magnesium sulfate, respectively, obtaining a delay in delivery of greater than 48 hours. Side effects were comparable in both groups, although they tended to be more serious in the patients receiving ritodrine. In patients receiving both drugs together, the rate of side effects was 77% without a demonstrable benefit over a single agent. We conclude that ritodrine and magnesium sulfate are tocolytics of comparable efficacy and when used aggressively are highly successful in delaying delivery. PMID- 3048102 TI - Low-dose aspirin therapy improves fetal weight in umbilical placental insufficiency. AB - A randomized, placebo-controlled, double-blind trial was carried out to evaluate the fetal benefits of low-dose aspirin (150 mg/day) as a treatment of placental insufficiency during the last trimester of pregnancy. Forty-six women referred for study because there was concern about fetal welfare were found to have an elevated umbilical artery wave form systolic/diastolic ratio. Mothers with severe hypertension were excluded because fetal condition would not necessarily be the dominant determinant of obstetric decision making. A distinction was made between a high systolic/diastolic ratio (greater than 95th but less than 99.95th percentile) and an extreme systolic/diastolic ratio (greater than 99.95th percentile). There were 34 patients in the high ratio group and 12 in the extreme group. Aspirin therapy was associated with an increase in birth weight (mean difference 526 gm [p less than 0.02]), head circumference (1.7 cm [p less than 0.025]), and placental weight (136 gm [p less than 0.02]) in those patients with a high initial umbilical artery systolic/diastolic ratio. For the 12 women with an extreme initial systolic/diastolic ratio, aspirin therapy did not result in a significantly different pregnancy outcome. PMID- 3048104 TI - 1938 to 1988, evolution of an idea: the next 50 years. PMID- 3048105 TI - Sensitivity and specificity of endocervical curettage and the endocervical brush for the evaluation of the endocervical canal. AB - A series of 87 consecutive conization specimens was studied to evaluate the accuracy of endocervical curettage for the detection of dysplasia in the endocervical canal and to investigate the role of the endocervical brush for the outpatient management of patients with atypical Papanicolaou smears. For patients having cervical intraepithelial neoplasia within the endocervical canal in conization specimens, the false-negative rate observed for endocervical curettage was 45% and the false-positive rate for the detection of endocervical involvement was 25%. The false-negative rate for the endocervical brush was only 8.4%, but the false-positive rate for endocervical involvement was 62.5%. The false negative rate of endocervical curettage could be reduced to 16.7% if an abundant volume of endocervical material (as determined by point counting) was required. The utility of endocervical curettage to detect cervical intraepithelial neoplasia in the endocervical canal appears to be dependent on the adequacy of the specimen. The determination of adequacy is a critical factor for the proper interpretation of specimens obtained by endocervical curettage. PMID- 3048106 TI - Continuous-wave Doppler ultrasound and decreased amniotic fluid volume in pregnant women with intact or ruptured membranes. AB - The cause(s) of decreased amniotic fluid in the absence of fetal anomalies and intrauterine growth retardation is not clear. A prospective study was performed to evaluate umbilical and uterine artery Doppler velocimetric results in pregnancies complicated by decreased amniotic fluid. Three medically high-risk groups were studied: women with (1) normal fluid and intact membranes, (2) decreased fluid and intact membranes, and (3) decreased fluid and ruptured membranes. The decreased fluid/intact membranes group had a significantly increased incidence of abnormal uterine artery waveforms (diastolic notching or absence of end-diastolic velocity); however, uterine systolic/diastolic ratios were not significantly different. The umbilical systolic/diastolic ratios were marginally higher in the intact membranes/decreased fluid group when compared with the ruptured membranes group. This study suggests that problems with maternal blood supply to the placenta may be related to decreased amniotic fluid when membranes are intact. PMID- 3048107 TI - Can a gynecologic Chlamydia trachomatis infection be diagnosed by direct observation of an endocervical plain slide? PMID- 3048108 TI - Pseudophakic bullous keratopathy. AB - We reviewed the records of all patients with pseudophakic bullous keratopathy (271 eyes, 251 patients) seen during a six-month period to determine predisposing factors, associated problems, current management, and visual outcome. Pseudophakic bullous keratopathy was associated most frequently with anterior chamber intraocular lenses in general (155 of 271), and with Leiske style lenses in particular (100 of 271). It was associated with a visual acuity of 20/200 or less in 206 eyes and a visual acuity of counting fingers or less in 129 of the eyes at the initial examination. Penetrating keratoplasties had been performed in 189 of the eyes. After penetrating keratoplasty, 108 of 189 of the eyes had a visual acuity of 20/200 or less (mean follow-up, 15 months). Visual acuity improved with longer follow-up, and among patients with a minimum follow-up of two years, 23 of 36 eyes had a visual acuity of 20/100 or better. Most grafts were clear (145 of 189). Pseudophakic bullous keratopathy was associated with marked visual loss, which was permanent despite clear grafts in 29 of 92 eyes followed-up for one year or longer. PMID- 3048109 TI - Enhancement of the ocular hypotensive effect of acetazolamide by diflunisal. AB - We studied the effect of diflunisal on intraocular pressure in patients with glaucoma who were receiving maximally tolerated therapy. Diflunisal therapy, 500 mg twice daily, was started in 48 patients for one week. No changes were made in their regular antiglaucoma medications. Intraocular pressure was reduced an additional 3.8 +/- 3.1 mm Hg (+/- S.D.) in the acetazolamide-treated patients (P less than .0001) and 1.6 +/- 1.5 mm Hg in methazolamide-treated patients (P less than .02), while no significant reduction in intraocular pressure was found in patients receiving topical medications alone. In 15 acetazolamide-treated patients, total plasma concentrations of acetazolamide after diflunisal therapy were significantly higher than the prediflunisal levels, suggesting a modest decrease in renal excretion. In seven acetazolamide-treated patients, free plasma concentrations of acetazolamide were found to increase 5.6-fold after diflunisal therapy. We concluded that diflunisal potentiated the ocular hypotensive effect of acetazolamide by increasing its free plasma level. PMID- 3048110 TI - Predicting the risk of hereditary retinoblastoma. PMID- 3048111 TI - Cyanide poisoning victims as corneal transplant donors. PMID- 3048112 TI - A technique for repairing strabismus after scleral buckling surgery. PMID- 3048113 TI - Effects of flow rate and insulin on triacylglycerol secretion by perfused rat liver. AB - In rat livers perfused with undiluted rat blood at perfusion rates of 6, 12, or 18 ml/min, hepatic O2 consumption rose with blood flow. Lipogenesis was unaffected by blood flow in control livers and was enhanced by insulin at 12 and 18 ml/min. Very-low-density lipoprotein triacylglycerol secretion also rose with increased flow and was stimulated by insulin at both 6 and 12 ml/min. When glucose was added to livers perfused at 12 or 18 ml/min, uptake was independent of perfusion rate and was slightly stimulated by insulin. Total lipogenesis and the secretion of newly synthesized fatty acids in very-low-density lipoprotein triacylglycerols were unaffected by insulin at either flow rate. The hormone stimulated triacylglycerol secretion at 18 ml/min but inhibited it at 12 ml/min. It seems that in perfused liver, effects of insulin on lipogenesis and very-low density lipoprotein secretion may be modified not only by changes in O2 consumption (in this case through alterations in blood flow) but also by the choice of substrate. PMID- 3048114 TI - Triton WR-1339 injected into rats enters renal renin granules. AB - To test directly the possibility that substances in extracellular fluid can gain access to renin storage granules in renal juxtaglomerular cells, rats were injected with Triton WR-1339, which binds to plasma proteins. A heavy granule fraction was prepared, and isopycnic sucrose density gradient centrifugation was performed. The renin granule peak was found to be altered from a mean equilibrium density of 1.202 g/ml in control rats to 1.196 g/ml for rats injected with Triton WR-1339 (P less than 0.005). The distribution of angiotensinogen, which is bound in kidney granules having a different buoyant density, was also examined and found to be unaltered. After injection, Triton WR-1339 binds to circulating plasma proteins. The results for renin support the possibility of pinocytotic uptake of protein-Triton WR-1339 complexes by the juxtaglomerular cells with subsequent fusion of the endocytotic lysosomal vacuoles with renin granules accounting for the translocation of ingested substances into the granule matrix. If so, the potential would therefore exist for interaction(s) of ingested extracellular substances with renin or other components in the granules. The present study has therefore demonstrated directly that endogenous extracellular substances may enter renin granules. PMID- 3048115 TI - Physiological hypercortisolemia increases proteolysis, glutamine, and alanine production. AB - Physiological elevations of plasma cortisol levels, as are encountered in stress and severe trauma, were produced in six normal subjects by infusing them with 140 micrograms.kg-1.h-1 of hydrocortisone for 64 h. Amino acid kinetics were measured in the postabsorptive state using three 4-h infusions of L-[1-13C]leucine, L [phenyl-2H5]-phenylalanine, L-[2-15N]glutamine, and L-[1-13C]alanine tracers 1) before, 2) at 12 h, and 3) at 60 h of cortisol infusion. Before and throughout the study, the subjects ate a normal diet of adequate protein (0.8 g.kg-1.day-1) and energy intake. The cortisol infusion raised plasma cortisol levels significantly from 10 +/- 1 to 32 +/- 4 micrograms/dl, leucine flux from 83 +/- 3 to 97 +/- 3 mumol.kg-1.h-1, and phenylalanine flux from 34 +/- 1 to 39 +/- 1 (SE) mumol.kg-1.h-1 after 12 h of cortisol infusion. These increases were maintained until the cortisol infusion was terminated (64 h). These nearly identical 15% increases in two different essential amino acid appearance rates are reflective of increased whole body protein breakdown. Glutamine flux rose from 325 +/- 28 to 453 +/- 28 mumol.kg-1.h-1 by 12 h of cortisol infusion and remained elevated at the same level at 64 h. The increase in flux was primarily due to a 55% increase in glutamine de novo synthesis. Alanine flux increased from 207 +/- 13 to 285 +/- 23 mumol.kg-1.h-1 with acute hypercortisolemia and increased further to 475 +/- 59 mumol.kg-1.h-1 at 60 h of cortisol infusion, a result primarily of increased alanine de novo synthesis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3048116 TI - James Braid's psychophysiology: a turning point in the history of dynamic psychiatry. AB - James Braid is both a neglected and integral figure in the history of dynamic psychiatry. With the introduction of his neurophysiologic theory of hypnosis in the early 1840s he buried Mesmer's doctrine of animal magnetism and established hypnotic phenomena as data suitable for scientific inquiry. He subsequently elaborated a sophisticated psychophysiology with emphasis on the psychology of suggestion and the phenomenon of double consciousness. His espousal of hypnotism as a tool of scientific investigation and his innovative use of hypnosis to cure hysterical paralysis profoundly influenced the debates of the 1880s and 1890s concerning suggestive therapeutics and the nature of hypnosis. PMID- 3048118 TI - Clinical and psychobiological characteristics of simultaneous panic disorder and major depression. AB - Simultaneous major depression and panic disorder appears to be a common occurrence in psychiatric patients. Patients with this condition present with more severe symptoms than patients with major depression only, respond less well to conventional antidepressants, and in general exhibit greater psychopathology over the course of their illness. Evidence suggesting a possible "dual diathesis," depression and panic, in these patients is reviewed from epidemiological, clinical, and biological perspectives. The importance of taking into account the combined symptoms in treatment planning and physiopathological studies is discussed. PMID- 3048117 TI - Blood-injury phobia: a review. AB - Natural human uneasiness about blood, injury, or deformity sometimes becomes a specific phobia, which can lead to serious disability if vital medical procedures are refused. Blood-injury phobia usually starts in childhood and is often familial. Unlike other phobic cues, which cause persistent tachycardia, blood injury phobic cues evoke an initial rise in heart rate followed by vasovagal bradycardia and, frequently, syncope. Although blood-injury phobia may have an evolutionary, genetic, and physiological basis, it can be treated effectively by exposure. The tendency to faint early in exposure therapy can be reduced by lying down, tensing the muscles, or inducing anger. PMID- 3048119 TI - Psychiatry and the resource-based relative value scale. AB - Attention to reform of reimbursement for psychiatric inpatient services largely focuses on the use of prospective payment systems, e.g., payment based on diagnosis-related groups (DGRs), for hospitals. Recently, there also has been interest in proposals for altering physician reimbursement (inpatient and outpatient) by using physician DRGs, capitation models, or relative value scales instead of the charge-based, fee-for-service model. The authors review the resource-based relative value scale (RBRVS) as an option for psychiatry. The RBRVS uses the setting, the time spent, the difficulty in treating the patient, the training, and the psychiatrist's role to determine reimbursement rates. PMID- 3048120 TI - An open trial of buspirone in obsessive-compulsive disorder. AB - Of 14 patients with obsessive-compulsive disorder who entered an 8-week open trial of buspirone, none improved. The ineffectiveness of buspirone may shed light on the serotonergic hypothesis that has been proposed for this disorder. PMID- 3048121 TI - A randomized clinical trial of phenelzine and imipramine for posttraumatic stress disorder. AB - In a double-blind, randomized clinical trial, the efficacy of imipramine and of phenelzine was compared with that of placebo in 34 male veterans with posttraumatic stress disorder (PTSD). Both medications reduced PTSD symptoms. PMID- 3048122 TI - Effects of trazodone on aggressive behavior in seven patients with organic mental disorders. AB - Seven patients with organic mental disorders and physically aggressive behavior were treated with trazodone. Three ceased demonstrating aggressive behavior within 4-6 weeks after starting trazodone treatment. Three had no discernible reduction in aggressiveness. One was unable to complete the trial. PMID- 3048123 TI - Lack of efficacy of pindolol in tardive dyskinesia. PMID- 3048124 TI - PTSD and carbamazepine. PMID- 3048125 TI - Emergency department patient 'dumping': an analysis of interhospital transfers to the Regional Medical Center at Memphis, Tennessee. AB - To study the extent and nature of transfers of emergency department (ED) patients because of inability to pay, we audited all telephone requests and actual patient transfers from private hospital EDs and their affiliated free-standing emergency centers to the ED of the Regional Medical Center at Memphis (the Med), a publicly subsidized hospital, between June 1 and August 31, 1986. Transfers to the Med's "special care" centers were assumed to represent tertiary care referrals and were excluded. During the 92-day study interval, ED physicians at the Med handled 190 telephone requests for transfer. Requesting physicians explicitly identified "no money" or "no insurance" as the primary reason for transfer in 89 per cent of 164 cases in which these data were recorded. Thirty-seven per cent of requests were refused; half were too unstable or required an intensive care unit (ICU) bed when none were available. One hundred forty-six transfers (55 per cent) arrived without prior telephone authorization, most by private automobile. Almost all transferred patients (91 per cent) were sent for primarily economic reasons. One out of four was found to be unstable on arrival by explicit clinical criteria. Eighty-two patients transferred for economic reasons (34 per cent) required emergency hospitalization and accounted for 564 bed days during a period of extreme inpatient crowding. Three patients died prior to discharge. Two had been transferred for primarily economic reasons. PMID- 3048126 TI - International developments in abortion laws: 1977-88. AB - During the period between 1977 and the first quarter of 1988, 35 countries liberalized their abortion laws and four countries limited grounds for the procedure. Most legislation has extended abortion eligibility through traditional indications such as danger to maternal health or fetal handicap, but a number of other indications have been created such as adolescence, advanced maternal age, family circumstances, and AIDS or HIV infection. A number of countries have redesigned their abortion laws as part of a comprehensive package to facilitate access to and delivery of contraception, voluntary sterilization, and abortion services. Abortion litigation has increased and stimulated the liberalization of abortion provisions and the support of women's autonomous choice within the law. In Canada, the entire criminal prohibition of abortion was held unconstitutional for violating women's integrity and security. In contrast, Latin American and other constitutional developments may limit legal abortion to instances of danger to women's lives. PMID- 3048127 TI - AIDS reference service at National Library of Medicine. PMID- 3048128 TI - Joseph B. DeLee and the practice of preventive obstetrics. PMID- 3048129 TI - Distribution of Trop 3 and 4 antigens in human endometrial glandular epithelium. AB - The reactivities of three monoclonal antibodies (MAbs), Trop 3, Trop 4, and W6/32, were studied on human uterine tissue by using an indirect immunoperoxidase method. Nonpregnant endometrial glandular epithelium was stained by all three MAbs. During pregnancy, Trop 3 expression was not detectable, but Trop 4 and W6/32 showed variable staining on the endometrial glandular epithelium, especially during late pregnancy. These findings support previous work suggesting a local regulation of antigenic expression by endometrial glandular epithelial cells. PMID- 3048130 TI - T and B lymphocyte populations in uterus draining lymph nodes at estrus and diestrus in Wistar rats. AB - In this paper, data are presented on the characterization of uterus draining lymph nodes (UDLN) B and T lymphocytes of virgin rats at estrus (E) and diestrus (D). We established that the T/B lymphocyte relationship in the UDLN is less than one at estrus and more than one at diestrus. This is due to a decrease in the percentage and in the total number of mature T cell population in association with a decrease in the percentage of the OX8 subset in the UDLN at estrus. This situation may be related to an increase in estrogens known to produce thymic involution. These changes were not observed when we studied peripheral (popliteal) lymph nodes. The changes observed in the UDLN T cell population at estrus could be under hormonal control and we think that this condition may be important to prepare the immune system for an eventual pregnancy. PMID- 3048131 TI - Evidence for active immunological regulation in prevention of testicular autoimmune disease independent of the blood-testis barrier. AB - It has long been considered that autoimmune disease of the testis is prevented by sequestration of testis-specific autoantigens on germ cells behind the blood testis (BT) barrier. However, we now have evidence that not all such antigens are sequestered. Some appear to reside on germ cells in the basal compartment of the seminiferous tubule where they are accessible to antibodies and to circulating activated T cells. Mice immunized with syngeneic testis homogenate are found to have immunoglobulin G (IgG) bound to cells in the basal compartment before onset of orchitis. This IgG is absorbed from circulation by the testis and, therefore, found only in the serum of mice orchiectomized before immunization. When the IgG is eluted from the testis, it is found to react preferentially with testicular cells enriched in preleptotene spermatocytes. T cells from mice immunized with testis can be transferred to naive syngeneic mice where they infiltrate the testis to cause orchitis. This implies that the BT barrier does not need to be breached directly for specific T cells to have access to testicular autoantigens on antigen presenting cells. Thus, active systemic and/or local immunoregulatory mechanisms must operate to prevent testicular autoimmune disease. These mechanisms may operate at the level of suppressor T cells, nonspecific suppression in the local environment of the testis, antigen presentation in the testis, or lymphocyte trafficking in the testis. These mechanisms probably operate only on the afferent limb of the immune response since they are overridden and orchitis occurs once testis-specific activated T cells are generated. PMID- 3048132 TI - Surgical management of the acutely obstructed colon. A review of 127 cases. AB - The surgical results in 127 cases of acute obstruction of the colon are presented. Carcinoma continues to account for the overwhelming number of cases, and there has been no appreciable change in the site of obstruction or age groups affected. In the current study, the overall mortality rate in patients with acute obstruction from all causes was 27 percent, which does not appear to be significantly different than it was 30 years ago. The overall mortality rate in patients with obstruction secondary to carcinoma was 23 percent. Under the specific circumstances of the cases reported herein, and on the basis of a limited experience, total colectomy and left colectomy as initial procedures in acute obstruction secondary to cancer had the same mortality rate as staged resection of the left colon. The only benefit found from either approach was an increase in the disease-free 5 year survival rate with staged resection. The overall survival rate was not enhanced by either approach. PMID- 3048133 TI - Who's afraid of the dentate line? The Whitehead hemorrhoidectomy. AB - Between January 1, 1970 and January 1, 1980, 1,002 hemorrhoidectomies, of which 356 were modified Whitehead procedures, were performed at a University of Illinois-affiliated hospital. Follow-up data were obtained from 295 of these patients, with times ranging from 5 to 12 years. Five patients experienced postoperative contractures. Of these, three required a second procedure and one required a third. There were no cases of fecal incontinence or rectal seepage. After review of the literature, I concluded that (1) the poor record of the Whitehead hemorrhoidectomy is due to the confusing and oftentimes misleading terminology of the anorectal anatomy found in early texts, and (2) this technique, or one of its modifications, is a viable treatment option for patients with circumferential, prolapsing, mixed hemorrhoids. PMID- 3048134 TI - Cyclosporin A and islet function. AB - Long-term cyclosporin A (CsA) administration in dogs was studied with respect to function of the islets of Langerhans. After 3 weeks of immunosuppression with therapeutic doses, the islets were isolated and assessed in vitro for insulin release in response to glucose challenge. Islet tissue retrieved from the CsA treated animals showed a total insulin output significantly lower than that of the control animals (p less than 0.01). The first and second phases of insulin release were both impaired in animals treated with CsA compared with controls (p less than 0.001 and p less than 0.05, respectively). The negative impact of CsA on the beta cells was easily demonstrated in this in vitro study. Similar results are more difficult to achieve with purely in vivo models, probably due to the great redundancy of the islet mass in intact animals. The mechanism of this CsA toxicity remains to be defined. PMID- 3048135 TI - Surgical disorders of the pancreas in infancy and childhood. AB - Pancreatic disorders in infants and children encountered over a 20 year period are reviewed. A total of 79 children were treated. Forty-eight had pancreatitis or its complications, 17 had congenital malformations, 12 had hypoglycemia and hyperinsulinism, and 2 had carcinoma. The mortality rate for the children with pancreatitis was 17 percent and was limited to patients treated nonoperatively. Idiopathic and drug-induced pancreatitis (the latter, particularly from corticosteroids) were the predominant types. Only rarely should such patients undergo operative treatment. Operations performed for various obstructive or traumatic lesions of the pancreas, as well as for complications of pancreatitis, obtained uniformly good results. The most common congenital malformation of the pancreas was an annular pancreas in association with duodenal atresia; all children with this abnormality were successfully treated with bypass procedures. Four patients with an ectopic pancreas underwent successful wedge resection. Nine infants with nesidioblastosis or islet cell hyperplasia and three children with islet cell adenomas underwent successful resection without any deaths, although neurologic sequelae due to prolonged preoperative hypoglycemia were common. Two patients underwent radical resection for pancreatic carcinoma, one of whom had survived 20 years postoperatively at last follow-up. Pancreatic disorders requiring operation in childhood are uncommon, but are likely to be complex and challenging when they do occur. PMID- 3048136 TI - A four-technique comparative study of orthotopic liver transplantation in the rat. AB - A four-technique controlled study was conducted on rat liver isografting to compare the microsurgical technique (Group 1), the two-cuff technique (Group 2), the three-cuff technique (Group 3), and a newly developed splinting suture technique (Group 4). The 60 day survival rates were significantly better in Groups 2 and 4 than in Groups 1 and 3. The anhepatic phase was significantly shorter with the cuff and splinting techniques than with the microsurgical technique, and total hepatic ischemic time was significantly shorter in the splint group than in the other three groups. Hepatic failure and shock had a higher incidence in those groups displaying a longer total hepatic ischemia time and a longer anhepatic phase. The results of this study show that the two-cuff technique and the splinting technique have substantial advantages over the microsurgical technique and the three-cuff technique. In particular, the splinting technique is preferable to the two-cuff technique because total hepatic ischemia time is shorter, and it is faster for a well-trained microsurgeon to learn this procedure than to learn the cuff technique de novo. PMID- 3048137 TI - Needle localization breast biopsy: accuracy versus cost. PMID- 3048138 TI - Duplex scanning for carotid artery disease: is angiographic confirmation required? PMID- 3048139 TI - Biliary lithotripsy. PMID- 3048140 TI - Endoscopic ligation of esophageal varices. PMID- 3048141 TI - [Ultrasonic examination of pregnant women with iron deficiency anemia]. PMID- 3048142 TI - [Epidemiology of hypertensive disorders in pregnancy]. PMID- 3048143 TI - [The course of pregnancy and the status of the feto-placental system in arterial hypotension]. PMID- 3048144 TI - [Relaxin and pregnancy]. PMID- 3048145 TI - Tracheal stenosis or agenesis in association with tracheo-oesophageal fistula and oesophageal atresia. AB - Two babies are described with oesophageal atresia, a tracheo-oesophageal fistula and severe subglottic tracheal stenosis. A third baby, who did not survive, had a tracheal agenesis associated with bronchi arising from the oesophagus. A review of the types of tracheal stenosis and agenesis associated with various forms of tracheo-oesophageal fistula is included. PMID- 3048146 TI - Pre-eclampsia in a parturient with a history of myocardial infarction. A case report and literature review. AB - A mother with pre-eclamptic toxaemia and severe coronary artery disease was managed with epidural analgesia accompanied by invasive cardiovascular monitoring. Caesarean section was carried out uneventfully using the epidural. The literature is reviewed. PMID- 3048147 TI - Clinical trial of the Continucath intra-arterial oxygen monitor. A comparison with intermittent arterial blood gas analysis. AB - An intra-arterial continuous display oxygen electrode for radial artery cannulation is now available in the UK. Nine catheters were used in patients during and after hypothermic cardiac surgery. Results obtained were compared with those from conventional intermittent blood gas samples. Slow temperature response times negate its use during hypothermic surgery. A good correlation was found during the postoperative period. Indications for its use are discussed. PMID- 3048148 TI - A simple CPAP system during one-lung anaesthesia. PMID- 3048149 TI - Survival time during hypoxia: effects of nitrous oxide, thiopental, and hypothermia. PMID- 3048150 TI - In vitro anesthetic washin and washout via bubble oxygenators: influence of anesthetic solubility and rates of carrier gas inflow and pump blood flow. AB - The uptake and elimination of volatile anesthetic agents administered to patients under conditions of hemodilution and hypothermia during cardiopulmonary bypass have not been determined. To define the limitations imposed by oxygenators, we defined washin and washout curves for volatile anesthetic agents administered to bubble oxygenators primed with diluted blood (without connection to a patient). There was rapid equilibration of anesthetic partial pressure between delivered gas and blood (85-90% within 16 minutes). Increasing the gas inflow to the oxygenator from 3 to 12 L/min hastened washin and washout slightly, while increasing the pump blood flow from 3 to 5 L/min had no effect. Rates of washin and washout of anesthetics differed as a function of their blood/gas solubilities: enflurane greater than isoflurane greater than halothane during washin; isoflurane greater than enflurane greater than halothane during washout. However, these differences were small. Oxygenator exhaust partial pressures of anesthetic correlated with simultaneously obtained blood partial pressures, suggesting that monitoring exhaust gas may be useful clinically. PMID- 3048151 TI - Extracranial carotid atherosclerosis evaluation and stroke occurrence: role of the echotomographic analysis. AB - High-resolution real-time echotomography was used in a longitudinal study on 118 atherosclerotic plaques of the extracranial carotid tract with the aim of identifying those most likely to cause a cerebrovascular event. Seventy patients (average age 61 +/- 7 years), referred to our Clinical Vascular Laboratory because suffering from transient cerebral ischemic attacks (TIA), coronary heart disease (CHD, peripheral artery disease (PAD), or cervical bruits (CB), were followed up for two years. Medical treatment remained that of the referring physician. Ten patients suffered from clinical events caused by stroke, TIA, and/or carotid occlusion during the follow-up. The echostructural profile of the lesions most often correlated with the clinical event was characterized by a mixed or hard echogenic pattern, and irregular surface, and an initial vascular stenosis of more than 50%. PMID- 3048153 TI - Efficacy of indobufen in the treatment of intermittent claudication. AB - The aim of this trial was to assess the activity of indobufen compared with placebo in peripheral occlusive arterial disease of the lower limbs of atherosclerotic or diabetic origin. Fifty-two outpatients were admitted to the randomized, double-blind study and were given either an indobufen 200-mg tablet (28 subjects) or placebo (24) for six months. Painfree walking distance on a treadmill at a constant speed (4 km/h) and slope (10 degrees) was assessed before and after three and six months' treatment. The painfree walking distance before treatment with indobufen or placebo averaged 153 +/- 23.02 (mean +/- SE) and 199 +/- 30.58 (mean +/- SE) meters respectively. After six months' treatment with active drug or placebo, this parameter reached 610 +/- 115.36 (p less than 0.01) and 243 +/- 32.49 (p greater than 0.05) meters respectively. The difference between the two treatments was statistically significant in favor of indobufen (p less than 0.01 Dunn's test). PMID- 3048152 TI - Are strokes more likely to result from severe carotid atherosclerotic stenoses? If not, why not? AB - The authors studied by carotid duplex ultrasonography 478 unselected elderly patients (age sixty to one hundred one years, mean 82 +/- 8), in their long-term health care facility. Of these, 108 had previously experienced strokes owing to atherothrombotic brain infarcts, documented by a neurologist, and 370 had not. The degree of extracranial internal or common carotid narrowing in these patients was classified by standard Vmax Doppler criteria as 0-40%, 40-80%, and 80-100% luminal diameter reduction. The authors found that 87% of the patients had little or no carotid stenosis (0-40% luminal diameter reduction). Nevertheless, 79% of the previous strokes had occurred in these patients, and the incidence of strokes in this group was 21%. Severe, but not mild or moderate, degrees of carotid obstruction (80-100% luminal diameter reduction) were associated with a 100% stroke incidence. The authors conclude that strokes most commonly result from causes other than ischemia due to in situ severe extracranial internal or common carotid stenosis or occlusion. PMID- 3048154 TI - The interdependence of hypertension, calcium overload, and coronary spasm in the development of myocardial infarction. AB - It is a well-known fact that systemic hypertension is one of the major risk factors for myocardial infarction (MI). Extensive studies on hypertensive rats revealed that calcium is excessively elevated in the myocytes, as well as in the coronary artery wall of these animals, which results in a higher resting tension and a stronger contractile response of those muscle strips. Over many years coronary spasm has been claimed by various authors to be greatly involved in the pathophysiology of early phase of acute MI (AMI). It can be shown that thrombocytes that aggregate at the injured vessel wall next to atherosclerotic plaques release vasoconstrictive factors that induce series of severe spasms at the sites with defective endothelium that end up in myocardial infarction; the pathophysiologic pathway is called the thrombo-ischemic reentry mechanism. This local contractile response may be enhanced in the presence of systemic hypertension since intracellular calcium is elevated in the coronary smooth muscle. On the other hand, it has been shown that heart muscle fibers undergo severe alterations finally resulting in necrotization, as soon as free calcium ions penetrate excessively through the sarcolemma membrane into the myoplasm so that the capacities of the calcium binding or extrusion processes become overpowered; this is especially the case during ischemia. Since free intracellular calcium is already ten times elevated in the myocytes in systemic hypertension, the myocardium may be more vulnerable to further calcium overload owing to the ischemia and necrotization is augmented. The elevation of intracellular Ca of the myocytes of the cardiovascular system in systemic hypertension enhances the pathologic response of the coronary arteries and the myocardium. This work gives a complete overview of the pathophysiologic principles involved in AMI occurring with systemic hypertension. PMID- 3048155 TI - Multicenter double-blind study of ticlopidine in the treatment of intermittent claudication and the prevention of its complications. AB - In this multicenter trial 169 patients with chronic intermittent claudication due to obstructive peripheral vascular disease were randomized in a double-blind fashion into two parallel groups receiving either 250 mg ticlopidine or placebo, twice daily. At entry, the two groups (83 ticlopidine, 86 placebo) were well matched for the major clinical features apart from an excess of women in the ticlopidine group. At six months, 167 patients were alive, 2 having died of malignant disease (1 from each group). At this stage, 39 patients from the ticlopidine group and 29 from the placebo group (p = 0.04) had increased their walking distance by more than 50% of baseline values. For the groups as a whole pain-free and total walking distance were greater in the ticlopidine group than in the placebo group (194 vs 124 meters, p = 0.03 and 236 vs 170 meters, p = 0.04, respectively). Two patients from the ticlopidine group vs 9 patients from the placebo group (p = 0.03) developed significant cardiovascular events during the study. These results indicate that ticlopidine has a beneficial effect both in the treatment of the symptoms and the prevention of vascular complications in patients with intermittent claudication. PMID- 3048156 TI - [Mass spectrometry of peptides]. AB - Recent progress in biochemistry has shown the occurrence of many important peptides, among antibiotics, immunostimulants, hormones and neuromediators. The mass spectrometric study of these components by classical ionization techniques such as electron impact, chemical ionization or field desorption require a prior chemical derivatization because of their amphoteric properties, their low volatility and low thermostability. Recent improvements in methods of ionization, fast atom bombardment, secondary ion mass spectrometry, or mass spectrometry/mass spectrometry have made it possible to study the structure of peptides with more than 15 amino acid residues and eased mass spectra interpretation. Through the different methodologies described in this review three characteristic informations can be obtained concerning the molecular mass of peptides, the nature of constituent amino acids and the peptidic sequence and even quantitative measurements can be performed. PMID- 3048157 TI - Postweaning diarrhea in swine: experimental model of enterotoxigenic Escherichia coli infection. AB - A reproducible model of postweaning colibacillosis was obtained by controlling management and environmental variables to simulate conditions often seen at weaning. Suckling pigs were exposed briefly to starter diet at 1 week of age, weaned at 3 weeks of age, held at an ambient temperature of 20 +/- 2 C, and again given the starter diet. One day after weaning, each pig was given 10(10) colony forming units of enterotoxigenic Escherichia coli strain M1823B (O157:K88ac:H43 LT+ STb+) in broth containing 1.2% sodium bicarbonate via stomach tube. In vitro adhesion by strain M1823B to isolated intestinal branch borders was used to test pigs for susceptibility to K88. In this model, 3 syndromes were induced in susceptible pigs: (1) peracute fatal diarrhea; (2) moderate diarrhea, weight loss, and fecal shedding of the inoculum strain; and (3) no diarrhea, weight loss, and fecal shedding of the inoculum strain. Rotavirus particles were not found in fecal specimens of pigs with diarrhea. The K88-susceptible, noninoculated control pigs remained clinically normal. It was concluded that susceptibility to adhesion by K88+ enterotoxigenic Escherichia coli was a requirement for the production of disease in this model; inoculation with rotavirus was not necessary. PMID- 3048158 TI - Postweaning diarrhea in swine: effects of oxytetracycline on enterotoxigenic Escherichia coli infection. AB - Investigators have found that oxytetracycline decreases the adhesion of K88+ Escherichia coli to intestinal epithelial cells in vitro. This occurs with oxytetracycline-sensitive E coli at drug concentrations less than those required to prevent growth and with E coli that are resistant to the drug. We conducted experiments to determine whether oxytetracycline alters the disease caused by an oxytetracycline-resistant K88+ enterotoxigenic strain of E coli. Oxytetracycline treated pigs (inoculated with K88+ E coli) did not differ from nontreated pigs in the incidence or severity of diarrhea, nor in the shedding of K88+ E coli. However, during recovery, weight gain by treated pigs was slower than that of nontreated pigs. The control pigs were not inoculated with E coli, and they remained clinically normal. Oxytetracycline-treated controls gained weight faster than nontreated controls. Some controls were genetically resistant to K88+ E coli, others were susceptible. The K88-resistant oxytetracycline-treated controls gained weight faster than the K88-susceptible oxytetracycline-treated and non treated controls. PMID- 3048159 TI - Treatment of sustained ventricular arrhythmias: which therapy to use? PMID- 3048160 TI - Myocarditis and endomyocardial biopsy in unexplained heart failure: a diagnosis in search of a disease. PMID- 3048161 TI - NIH conference. Cystinosis: progress in a prototypic disease. AB - OBJECTIVE: To review the history, basic defect, pathogenesis, clinical manifestations, diagnosis, and treatment of nephropathic cystinosis. DESIGN: Lysosomal membrane transport studies, clinical reports, and a historically controlled 7-year trial of oral cysteamine therapy. SETTING: University centers in the United States and Canada. PATIENTS: One hundred forty-eight children, aged 0 to 12, with nephropathic cystinosis before renal transplant, who had renal tubular Fanconi syndrome, failure to grow, corneal cystine crystals, and elevated leukocyte cystine; 34 patients, aged 9 to 29, after transplant, some with visual impairment, corneal erosions, pancreatic dysfunction, or neurologic deterioration. INTERVENTION: Before transplant, replacement of renal losses, and treatment with oral cysteamine (55 mg/kg body weight.d for 1 to 6 years) and topical cysteamine eyedrops (0.1%, 1 drop/h while awake, for 6 months). After transplant, oral cysteamine and symptomatic treatment of late complications. MEASUREMENTS AND MAIN RESULTS: Untreated patients reached renal failure at age 10. Oral cysteamine lowered leukocyte cystine over 80%, and in patients before transplant, improved growth and preserved renal function (mean creatinine clearance [+/- SE], 0.64 +/- 0.04 mL/s.1.73 m2 [38.5 +/- 2.5 mL/min.1.73 m2] in the cysteamine group compared with 0.50 +/- 0.03 mL/s.1.73 m2 [29.7 +/- 2.0 mL/min.1.73 m2] in controls; 95% CI for the difference, 1.8 to 15.8). Cysteamine eyedrops cleared the corneal crystals of two children less than 2 years old. CONCLUSIONS: Cystinosis is a lysosomal storage disease due to impaired transport of cystine out of lysosomes. In young children, growth can be improved and renal deterioration delayed or prevented by oral cysteamine. Nonrenal complications after transplant might be prevented with long-term oral cysteamine. PMID- 3048162 TI - Discovery and disquiet: research on the brain-dead. PMID- 3048163 TI - Ambulatory electrocardiographic monitoring: the test for ischemia in 1988? PMID- 3048164 TI - Edward J. Huth to retire: the search begins for his successor. PMID- 3048165 TI - Efficacy of cardiac rehabilitation services. With emphasis on patients after myocardial infarction. AB - During the 1970s, emphasis increased in clinical practice on early ambulation and exercise-based rehabilitation after myocardial infarction and other cardiac illnesses or procedures. This shift was based on the belief that exercise and improved conditioning would improve prognosis. We examine the evidence supporting this assertion. Most of the reports on cardiac rehabilitation are about patients who have coronary artery disease and a history of myocardial infarction. The review, therefore, is focused primarily on the patient who has had a myocardial infarction. Effects of cardiac rehabilitation, emphasizing exercise treatment and conditioning, are reviewed with regard to patient outcomes, including changes in functional (work) capacity, psychosocial functioning and health-related knowledge, risk factor modification, morbidity and mortality, and cardiac function. The safety of cardiac exercise programs is reviewed, and the use of telemetry monitoring is considered. We also discuss the role of cardiac rehabilitation in categories of patients other than those with myocardial infarction and the application of newer approaches to rehabilitation such as programs based in the patient's home. PMID- 3048167 TI - The impact of forensic issues on women's rights. PMID- 3048168 TI - A historical perspective on SIDS research. PMID- 3048166 TI - Sexual aggression in the great apes. AB - Species-typical frequencies of copulation during the menstrual cycle differ among common chimpanzee, orang-utan, and gorilla, but all three species exhibit a midcycle enhancement associated with estrus. Thus, in the natural habitat, chimpanzees mate for 10-14 days, orang-utans for 5-6 days, and gorillas for 2-3 days. In traditional laboratory pair-tests, however, conducted in a single cage with both animals freely accessible to each other, all three species of great apes copulate more frequently than the species-typical pattern. In all three species, moreover, the increased copulation appears to result from increased male sexual initiative (aggression), male dominance over females, and the inability of the female to avoid or escape from the male within the limited spatial conditions of the free-access test. This interpretation is supported by studies using restricted-access tests in which females control sexual access. These data suggest that male sexual aggression in our closest biological affiliates commonly occurs when females are rendered vulnerable to the male by the absence of the normal social constraints and spatial prerogatives typical of the natural habitat. The possible implications of this interpretation for a biological perspective on human sexual aggression are considered. PMID- 3048169 TI - Risk factors for SIDS. Results of the National Institute of Child Health and Human Development SIDS Cooperative Epidemiological Study. PMID- 3048170 TI - Sleep and cardiac arrhythmias. PMID- 3048171 TI - Airway reflexes and the control of breathing in postnatal life. PMID- 3048172 TI - Periodic breathing. PMID- 3048173 TI - Pathophysiology of sudden upper airway obstruction in sleeping infants and its relevance for SIDS. PMID- 3048174 TI - Mechanisms for abnormal apnea of possible relevance to the sudden infant death syndrome. PMID- 3048175 TI - Cardiorespiratory interactions in heart-rate control. PMID- 3048177 TI - Intrathoracic petechial hemorrhages: a clue to the mechanism of death in sudden infant death syndrome? AB - (1) Intrathoracic petechiae are characteristic of most SIDS cases, and tend to be more numerous in these cases than in deaths from other causes, including mechanical asphyxia. (2) Their localization suggests that intrathoracic negative pressure plays a role in their genesis. (3) Several human and animal studies suggest that petechiae arising from the pulmonary circulation may differ from those originating from systemic vessels in the thorax. (4) Experimental studies suggest that vigorous respiratory efforts are responsible for their formation. This would seem to exclude respiratory paralysis as a mechanism for most SIDS deaths. (5) These petechiae are not consistent with cardiac arrest or failure as the primary agonal event in SIDS. (6) The fact that petechiae are more prominent in SIDS than in other deaths adds to the evidence that SIDS is not a "wastebasket" diagnosis, despite the imperfection of existing diagnostic criteria. (7) These petechiae do not prove that the final mechanism of most SIDS deaths is upper airway obstruction. However, they do provide substantial support for that thesis. PMID- 3048176 TI - Cardiorespiratory control during sleep. PMID- 3048178 TI - The role of sleep and arousal in SIDS. PMID- 3048179 TI - Problems in management of infants with an apparent life-threatening event. PMID- 3048180 TI - [Anisakids and human anisakiasis. 1. Bibliographic data]. AB - Currently available data about larvae of Anisakidae (genera Anisakis, Pseudoterranova, Hysterothylacium and Contracaecum) from marine fishes are analysed. Hazard of such parasites for fish-consumers are emphasized. PMID- 3048181 TI - Forearm split-skin donor sites: are they cosmetically acceptable? AB - The cosmetic end result was assessed in a small series of volar forearm donor sites taken under identical conditions. In most cases the donor site remained paler. This was cosmetically acceptable to all patients except 2 young women. PMID- 3048182 TI - Fasciocutaneous V-Y advancement flap for repair of sacral defects. AB - Sacral defects from 6 to 11 cm in diameter were closed with bilateral fasciocutaneous V-Y advancement flaps. All the defects were easily closed without any postoperative complication. We believe that the V-Y advancement technique using the fasciocutaneous unit has some major advantages for repair of moderate size sacral defects. It is a safe, simple, and less invasive procedure. PMID- 3048183 TI - Anatomic investigations of nerves at the wrist: I. Orientation of the motor fascicle of the median nerve in the carpal tunnel. AB - The orientation of the motor fascicle of the median nerve in the carpal tunnel was investigated in dissections of 50 hands. Topographically, the motor branch was located on the radial-volar aspect of the median nerve in 60% of the hands, the central-volar aspect in 22%, and between these two locations in the remaining 18%. In 56% of the hands, the motor branch passed through a separate distinct fascial tunnel before entering the thenar muscles. Awareness of these patterns will facilitate appropriate surgical management of thenar muscle weakness or wasting associated with the carpal tunnel syndrome. PMID- 3048185 TI - Reconstruction of postburn female breast deformity. AB - Postburn breast deformity is a sequelae of severe scar contraction of the burned chest. During the past four years, 6 female patients with such a deformity required reconstruction, and the surgery was performed in our department. Three different types of breast reconstruction were undertaken, each one achieving a satisfactory aesthetic result. PMID- 3048184 TI - Clinical use of a tissue expander--enhanced transposition flap for face and neck reconstruction. AB - Tissue expanders have been used in reconstructive surgery with increasing frequency, primarily to construct local advancement flaps of tissue immediately adjacent to a tissue defect or deformity. These flaps often lack adequate mobility to allow coverage of large areas. This report describes the use of tissue expanders to enhance the area and vascularity of shoulder skin to provide suitable, ample tissue that can then be used as large pedicled transposition flaps for reconstruction of the face and neck. It is a clinical study based on previous laboratory studies cited that demonstrates that large flaps with very narrow pedicles remain well vascularized and can be transposed to cover very large defects. The tissue expander appears to enhance the vascularity of the flap. The thinning of the dermis and subcutaneous tissue seems to make the donor skin more similar in quality to face and neck skin. These studies suggest that even larger flaps may be developed to supply tissue to resurface the entire face or very large portions of it. PMID- 3048186 TI - Rapid tissue expansion in the treatment of myelomeningocele. AB - In 2 patients undergoing spinal fusion for correction of kyphoscoliosis secondary to myelomeningocele skin coverage was achieved with rapid tissue expansion. Tissue necrosis caused by overly rapid expansion in the first patient was prevented by rapid, but more controlled, expansion in the second patient. Rapid tissue expansion in the paraplegic in a carefully controlled setting is useful and can be done without unduly prolonging hospitalization. This technique has potential uses in other clinical situations as well. PMID- 3048187 TI - [Hormonal modifications induced by food intake contribute to the regulation of the body weight and to metabolic variations]. AB - In the rat the energy content of the body is subject to homeostatic control. The level of energy content depends on internal and external conditions. In the adult individual food intake is matched exactly to energy expenditure. Food intake leads to profound alterations in several hormonal plasma levels affecting metabolism e.g. insulin, glucagon and catecholamines. These hormones play an important feedback role in the storage and depletion of reserve tissues such as liver and fat cells. In this paper it is discussed that messages reporting energy content of the body may influence feeding motivation and that the CNS and especially the hypothalamus plays a directing role in this respect. Among these motivation setting messages the pancreatic hormones insulin and glucagon may be prominent factors in respectively long- and short-term regulation of food intake. PMID- 3048188 TI - [Peptides of digestive system and brain. Model of the cholecystokinin]. AB - The systemic administration of exogenous cholecystokinin (CCK), (a peptide released into the circulation when nutrients arrive in small intestine) provokes inhibition of food intake in numerous animal species and a sensation of satiation in humans. The mechanism involved requires participation of vagal afferent neurons to transmit information, possibly on gastric distension, to brain stem. Two crucial questions remain unanswered: does peripheral endogenous cholecystokinin transport information necessary for satiation at end of normal meals and if so, how and when is this message coded and deciphered; other studies have demonstrated that very small doses of exogenous cholecystokinin (too low to induce satiation if administrated peripherally) will induce satiety if injected into cerebral ventricles or directly into cerebral parenchyma. This suggest but does not prove that cerebral endogenous CCK also plays a role in satiety. It is not known how the satiating action of centrally-administered CCK is related to that of peripherally-administered CCK. Current studies aim at establishing a coherent schema of neuro-endocrine mechanisms implicated (from the gastrointestinal wall via the brain to a motivated behaviour) in the fact that food taken during a meal induces the end of the latter. PMID- 3048189 TI - [Regulation of body weight and food palatability]. AB - It is as though the body weight, or a variable correlated closely with body weight, was submitted to regulation. This regulation is analyzed from known human studies. This short bibliography study reviews interrelations between palatability of food stuffs and ponderostat. Findings show that palatability elevates ingestion rate, metabolic output rate and ponderostat set point. Reciprocally, ingestion and the set point modulate food palatability. PMID- 3048190 TI - [Endogenous opiates, palatability and control of food intake]. AB - Endogenous opiate or opioids are peptidic neuromodulators which bind with specific receptors (mu, kappa, delta, etc), and thus control numerous physiological process, mainly pain and food intake (FI). Endogenous or exogenous agonists of opioid receptors increase FI. Antagonists decrease it. However several results indicate discrepancies. The anorectic activity of antagonists is different of the activity of classical anorexigenic drugs since inhibition of opioid receptors did not induce long term change of body weight. The analysis of the neurophysiological action of the opioid system on FI can explain this apparent discrepancy. Opioids act on the short term control of FI modulating the rewarding properties of alimentary stimuli: agonist increase and antagonists decrease it. Such an action correspond to a control of food palatability or, in human, to a control of the sensory pleasure/displeasure associated with food stimuli. Opioid system does not seem to directly act on body weigh regulation. PMID- 3048191 TI - [Feeding behavior disorders in man]. AB - Diagnosis of anorexia nervosa is easily made using symptom lists. It is mandatory to check that the weight loss is due to no somatic cause. The prevalence of bulimia and syndromes associating bulimia and anorexia is increasing considerably. In these latter syndromes, while not in typical anorexia, major affective disorders are often found. Endocrinological abnormalities are numerous. However the part of denutrition itself in these abnormalities is not clear. Pathogenesis of these eating disorders remains mysterious. But most probably, the endogenous opioid system is involved, at least in their perpetuation. PMID- 3048192 TI - [Monoclonal antibodies]. PMID- 3048193 TI - [Physiology of food intake and regulation of body weight]. AB - Feeding is often influenced by competitive needs or by environmental factors. Nevertheless, in physiological conditions hunger is elicited by depletion of macronutrients. The first question is what specific signals cause food intake? Recent data suggest that the signal comes not from exclusive depletion of carbohydrates or of anyone type of the major macronutrients but from overall decreased cellular power production (hypoischymetric signal). Locomotion free metabolic rate has been shown to decrease preceding hunger and to increase in order to determine satiety. Satiation, which occurs largely before ingested nutrients can cross the intestinal barrier and replete the inner milieu, obeys a specific mechanism. Following the stimulation by ingestion of the oro-gastro intestinal receptors metabolic hormones such as glucagon and insulin are released and enhance metabolism of endogenous reserves. The so-induced endogenous "meal" brings about an anticipatory hypermetabolism which inhibits further ingestion. Post-absorptive satiety subsequently will replace the pre-absorptive state of satiation. The long term regulation of body weight is more subtle. When the actual body weight exceeds its defended value, responsiveness towards signals of hunger diminishes. This hyporesponsiveness seems to be a consequent of an enlargement of the microscopic fat depot at the level of hypothalamic receptive structures which parallels the peripheral increase of the macroscopic fat depot. Thus, hunger promoting signals become less effective and feeding decreases. A symmetrical process takes place in case of body weight loss which results in central hyperresponsiveness to hunger related stimuli and thus facilitates feeding and body weight recovery. PMID- 3048194 TI - [Lipogenesis, lipolysis and feeding rhythms]. AB - Recordings of rat's diurnal ad libitum feeding patterns and responses to short term food deprivation suggested that analysis of diurnal feeding rhythms could provide a clue for understanding mechanisms involved in control and regulation of food intake. A simultaneous recording of free feeding patterns and of respiratory exchanges showed that a nocturnal positive energy balance and hyperphagia were mainly due to a diversion of ingested nutrients to fat deposition while negative energy balance and hypophagia during daytime were an effect of fat mobilization and oxidation. Further it was demonstrated that a neuroendocrine diurnal cycle underlined the lipogenesis-lipolysis cycle with hyper-insulinism and glucose tolerance at night, hypoinsulinism and glucose intolerance during the day. Similar phenomena were found along with the human scheduled feeding. Negative correlations between nocturnal lipogenesis and subsequent daytime lipolysis and the diurnal cycle of free fed rats compared to experimentally induced and reversible obesity and to seasonal cycles of hibernators give evidence on the nature of the lipostatic mechanism and its monitoring by hypothalamic ventromedial nuclei. PMID- 3048196 TI - [Hemorrhoid prolapse]. PMID- 3048195 TI - [Magnetic resonance imaging of cancer of the rectum]. AB - MRI (Magnetic Resonance Imaging) has become a major diagnostic method, in many fields. Its results, in the pre-operative evaluation of rectal cancers, are presented here. The possibilities of determining by MRI, parietal extension, peri rectal fat invasion, extension to adjoining organs and nodes, are specified. Then, the contribution of MRI to the measurement of the distance between the lower pole of the tumor and the levator muscles plane, is evaluated. PMID- 3048198 TI - [Hemorrhagic rectocolitis. General management]. PMID- 3048197 TI - [Hemorrhagic rectocolitis. The ileo-anal pouch anastomosis represents considerable progress]. PMID- 3048199 TI - [Sinusoid dilatation and the use of oral contraceptives. Apropos of a case with a review of the literature]. AB - We are reporting the case of a 23 year-old woman who developed an acute painful syndrome of the right hypochondrium with hepatic cytolysis while taking oral estro-progesterone medications. The liver biopsy showed a sinusoid dilatation. The course was favorable after discontinuation of oral contraceptives. PMID- 3048200 TI - [Saturnine colic and hemolytic anemia due to ingestion of solid lead. Apropos of a case]. AB - We report a case of lead poisoning resulting from the ingestion of solid metallic lead. A 24 year old patient was admitted for abdominal pain and microcytic anemia. Review of the literature suggests that clinical manifestations of chronic poisoning by solid lead in adults are extremely rare. Their misreading can lead to erroneous diagnose. PMID- 3048201 TI - [Liver pathology in Wilson's disease]. PMID- 3048202 TI - [The value of ultrasound-guided cyto-biopsy in the study of livers with abnormal ultrasonograms]. AB - This retrospective study considers the diagnostic reliability of 97 cyto-biopsies performed with a fine needle and guided by ultrasonography in 92 patients with one or several suspicious liver lesions. The results of the cyto-biopsy were compared with the final diagnosis obtained by histological examination or by the evolution. In 65 confirmed malignant tumors, fine needle biopsy was concordant in 54 cases. The sensitivity of this method in the diagnosis of malignant liver lesions was 83% with a specificity of 93%. In 54 malignant cytologies, the differentiation between primary or secondary lesions was possible in 31 cases (57%). In 32 confirmed benign lesions, fine needle biopsy was concordant in 30 cases. This study confirms the advantage of cyto-biopsy guided by ultrasonography in the positive and etiological diagnosis of malignant liver tumors. PMID- 3048203 TI - Phage-receptor on the cell wall of Veillonella rodentium. AB - Veillonellophage N2 prevented from adsorbing to Veillonella rodentium ATCC 17743 cells treated with polymyxin B, and also to lipopolysaccharides (LPSs) of the host cells treated with antibiotics. Therefore, these results indicate that receptor to phage N2 is cell wall LPSs. The LPSs of V. rodentium ATCC 17743 cells as receptor were characterized. Lipid A and total carbohydrate accounted for approximately 40% of the weight of the lipopolysaccharide complex. Heptose and 2 keto-3-deoxyoctonate were also present. Amino compounds included glucosamine, galactosamine, and glycine. PMID- 3048205 TI - Oxygen and nitrate reduction kinetics of a nonflocculating strain of Zoogloea ramigera. AB - The oxygen and nitrate reduction kinetics of a nonflocculating strain of Zoogloea ramigera were determined. Axenic, nitrate-reducing bacterial suspensions were acclimated to various oxygen levels in a chemostat while measuring nitrate reduction in the presence of high ammonium nitrogen concentrations. Significant nitrate reduction was observed at oxygen concentrations up to 8 mg L-1. Oxygen consumption was inhibited by oxygen concentrations in excess of 2 mg L-1. PMID- 3048204 TI - The effect of growth conditions on production and excretion of extracellular antigens by three ascomycetous yeasts. AB - Ascomycetous yeasts produce extracellular antigens that are almost specific for the species. The antigen production by Hansenula wickerhamii and Stephanoascus ciferrii was independent of the carbon source and was proportional to the final cell density of the cultures. The same was true of chemostat cultures of Stephanoascus ciferrii, irrespective of the dilution rate and whether glucose or ammonia was the limiting nutrient. In cultures of Saccharomyces cerevisiae, however, antigen excretion mainly took place in the late exponential growth phase. Large amounts of antigen were extracted from the cell wall of Saccharomyces cerevisiae. A small amount was detected in the cytoplasm. PMID- 3048206 TI - Anaerobic degradation of organic compounds at high salt concentrations. AB - A number of obligately anaerobic fermentative bacteria are known to degrade a variety of organic substrates such as sugars, amino acids, and others, in the presence of high salt concentrations (up to 3-4 M) to products such as hydrogen, CO2, acetate and higher fatty acids, and ethanol. Our understanding of the fate of these products in hypersaline environments is still extremely limited. The occurrence of bacterial sulfate reduction is well established at salt concentrations of up to 24%; however, the bacteria involved have not yet been isolated in pure culture, and the range of electron donors used is unknown. Halophilic or halotolerant methanogenic bacteria using hydrogen/CO2 or acetate as energy source are notably absent; methanogenesis under hypersaline conditions is probably limited to such substrates as methanol and methylamines, which cannot be expected to be major products of anaerobic degradation of most organic compounds. PMID- 3048207 TI - Ultrasonographic transvaginal ovum retrieval. A new approach to in vitro fertilization. PMID- 3048209 TI - Photodynamic laser therapy. A bladder cancer protocol. AB - This data, along with data from other research studies, indicate that PDT is a promising new treatment for noninvasive bladder cancer. In the current study at Veterans Administration Medical Center, three of the four patients who received Thiotepa have had recurrences of their bladder tumors or positive urine cytology. The three patients who received PDT have not experienced any recurrences or positive urine cytology. Since we have been involved in this trial, we have developed some recommendations that other institutions may want to follow. The tip of the laser fiber is very bright. To decrease eye strain and fatigue, a lens cap with a green eye shield should be used on the cytoscope. All staff should wear green-colored safety eye glasses. This will protect their eyes from constant exposure to the intense incandescent radiation, and if the fiber should break, it would protect their eyes from the bright red color. With all the extra equipment such as ultrasound, power meters, light sources, and anesthesia equipment, it is important to avoid tripping over extension cords. The cords should be covered. Also, it is important not to bump into the equipment, which can happen because the OR is dark. At this time, the laser technician is the only person authorized at our medical center to operate the argon-pumped tunable dye laser. In the future, if more research programs are started or therapy is approved for other uses, additional personnel will be trained. PMID- 3048208 TI - Discharging outpatients. Factors nurses consider to determine readiness. PMID- 3048210 TI - Historical development of neural transplantation. AB - The techniques of neural transplantation are almost 100 years old. As these techniques begin to be used to treat human neurological disorders, it is important to remember the contributions of the many investigators who have advanced this fascinating area of neurobiology. PMID- 3048211 TI - Immunology of transplantation in the central nervous system. AB - The brain has long been considered an immunologically privileged site. Tissue transplanted to the central nervous system (CNS) is immunologically better tolerated than grafts to other regions of the body. With improved graft survival, tissue transplantation may provide new treatment options for previously incurable CNS disorders. The normal immune response is reviewed, followed by a discussion of the factors responsible for graft rejection. The modification of these factors to allow successful CNS transplantation is discussed. PMID- 3048212 TI - Intraoperative electrical stimulation of the brain in patients with obsessive compulsive neurosis. AB - Twenty patients with intractable obsessive-compulsive neurosis were operated under local anesthesia. Each patient had a lesion produced in 1 of the 4 brain targets: anterior internal capsule, rostral cingulum, middle cingulum, and genu of the corpus callosum. Before destructive permanent lesions were produced, the target area was stimulated electrically. Stimulation gave subjective or objective reactions in 30% of the patients: diminished anxiety, 3 patients; increased anxiety, 1 patient, and motor responses, 2 patients. None experienced an obsessive reaction to stimulation. The clinical effect of surgery was usually good. PMID- 3048213 TI - Purification and properties of sulfhydryl oxidase from bovine pancreas. AB - Immunofluorescent studies showed that antibodies prepared against bovine milk sulfhydryl oxidase reacted with acinar cells of porcine and bovine pancreas. A close inspection of the specific location within bovine pancreatic cells revealed that the zymogen granules, themselves, bound the fluorescent antibody. Bovine pancreatic tissue was homogenized in 0.3 M sucrose, then separated into the zymogen granule fraction by differential centrifugation. The intact zymogen granules were immunofluorescent positive when incubated with antibodies to bovine milk sulfhydryl oxidase, and glutathione-oxidizing activity was detected under standard assay conditions. Pancreatic sulfhydryl oxidase was purified from the zymogen fraction by precipitation with 50% saturated ammonium sulfate, followed by Sepharose CL-6B column chromatography. Active fractions were pooled and subjected to covalent affinity chromatography on cysteinylsuccinamidopropyl-glass using 2 mM glutathione as eluant at 37 degrees C. The specific activity of bovine pancreatic sulfhydryl oxidase thus isolated was 10-20 units/mg protein using 0.8 mM glutathione as substrate. Ouchterlony double-diffusion studies showed that antibody directed against the purified bovine milk enzyme reacted identically with pancreatic sulfhydryl oxidase. The antibody also immunoprecipitated glutathione-oxidizing activity from crude pancreatic homogenates. Western blotting analysis indicated a 90,000 Mr antigen-reactive band in both bovine milk and pancreatic fractions while sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed a single silver-staining protein with an apparent Mr 300,000. Thus, we believe that sulfhydryl oxidase may exist in an aggregated molecular form. Bovine pancreatic sulfhydryl oxidase catalyzes the oxidation of low-molecular-weight thiols such as glutathione, N-acetyl-L-cysteine, and glycylglycyl-L-cysteine, as well as that of a high-molecular-weight protein substrate, reductively denatured pancreatic ribonuclease A. PMID- 3048214 TI - Tinea capitis. Current concepts. PMID- 3048215 TI - Orthotopic transplantation during early infancy as therapy for incurable congenital heart disease. AB - Since November 1985, 14 neonates and young infants have undergone orthotopic heart transplantation at Loma Linda University Medical Center (LLUMC) as therapy for hypoplastic aortic tract complex. Eleven (78%) survived surgery and are living and well today. Three perioperative deaths resulted: one due to perforated peptic ulcer, one due to necrotizing pneumonitis, and one due to graft failure unrelated to rejection. No late deaths occurred in the 1-29 months of follow-up, during which time noninvasive surveillance techniques were used. Immunosuppression was accomplished using cyclosporine and azathioprine. Steroids and antithymocyte globulin were used for identified rejection episodes only. Ordinary childhood infections were tolerated well. All survivors were normotensive. There was no late renal dysfunction. Although inadequate donor resources remain a significant limiting factor for transplantation therapy during early life, these results suggest that cardiac transplantation is effective therapy for selected neonates and young infants with incurable congenital heart disease. PMID- 3048216 TI - Artificial dermis for major burns. A multi-center randomized clinical trial. AB - This communication presents an 11-center prospective randomized trial using the artificial dermis invented by Burke and Yannas. Patients with life-threatening burns who underwent primary excision and grafting within 7 days of injury had comparable sites randomized to receive either the artificial dermis (study site) or the investigator's usual skin grafting material (control site). Control materials were autograft, allograft, xenograft, or a synthetic dressing. Epidermal grafts were applied to the study site during a second operation, and surviving patients were followed for 1 year after grafting. One hundred thirty nine sites on 106 patients were studied. Mean burn size was 46.5 +/- 15% mean total body surface (TBSA). Overall mortality was 13%, and mean hospital stay was 68 +/- 45 days. Median artificial dermis take was 80% compared with 95% for all comparative sites, but the take was equivalent to that of all nonautograft control materials. Results with the artificial dermis improved slightly as the investigators became more familiar with the material. Donor site thickness for the study site averaged .006'' +/- .002'' compared to .013'' +/- .018'' for control (p less than .0001) and the epidermal donor site healed an average of 4 days sooner (10 +/- 6 vs. 14 +/- 8 days) (p less than .0001). As the wounds matured during the first year, both patients and surgeons felt that both sites became more comparable in appearance and function. At the completion of the study, there was less hypertrophic scarring of the artificial dermis, and more patients preferred the artificial dermis to the control graft. Artificial dermis with an epidermal graft provides a permanent cover that is at least as satisfactory as currently available skin grafting techniques, and uses donor grafts that are thinner and donor sites that heal faster. PMID- 3048217 TI - Immunologically mediated disease of the airways after pulmonary transplantation. AB - Obliterative bronchiolitis has occurred in eleven of 30 recipients of cardiopulmonary allografts who survived at least 4 months after transplantation, has caused significant morbidity, and has been associated with four of eleven late deaths in this series. Although some improvement, or at least stability, of pulmonary function has followed augmented immune suppression, it appears that once the process is recognized clinically, much of the damage to the airways is irreversible. The histopathology, response to therapy, and, most important, the response of donor specific alloreactivity in the lymphocytes from the lung (bronchoalveolar lavage and peripheral blood) suggest immune- mediated basis for bronchiolitis obliterans. The presence of donor specific alloreactivity detected by primed lymphocyte testing predicted obliterative bronchiolitis in five of six recipients (83% sensitivity, 91% specificity) was absent in ten of eleven recipients who have not as yet developed the process (negative predicted value of 91%). Currently, the presence of a positive primed lymphocyte test in the bronchoalveolar lavage of the cardiopulmonary recipient is an indication for early treatment by augmented immune suppression. PMID- 3048218 TI - [Isolation and immunoenzyme determination of human beta 2-microglobulin]. AB - Two procedures for isolation of homogeneous beta 2-microglobulin from urine of patients with systemic lupus erythematosus were developed: a procedure for isolation of beta 2-microglobulin with two-stage gel chromatography on Sephacryl S-200 and a procedure for isolation of homogeneous beta 2-microglobulin with affinity chromatography on rabbit polyclonal antibodies. The microglobulins isolated with the two procedures showed identical physicochemical properties and were used in development of a competitive immunoenzymatic assay method for determination of beta 2-microglobulin in the blood. The method was approved for determination of beta 2-microglobulin in blood serum of patients. PMID- 3048219 TI - [Reasons for discrepancies in laboratory and clinical data on the antibiotic sensitivity of gram-negative bacteria]. PMID- 3048220 TI - [Lactobacillus phages and plasmids]. PMID- 3048221 TI - Comparison of the response to angiotensin II of isolated dog coronary and mesenteric arteries of proximal and distal portions. AB - Helically cut strips of dog coronary and mesenteric arteries of proximal and distal portions contracted in response to angiotensin (ANG) II in a dose dependent fashion. The contractions were greater in the distal portions than in the proximal portions. Mesenteric arteries responded to the peptide with a greater contraction than coronary arteries. The peptide-induced contraction was suppressed by treatment with saralasin and was potentiated by indomethacin; magnitudes of the potentiation were similar in coronary and mesenteric arteries and in proximal and distal portions. Concentrations of TRK-100, a stable analog of PGI2, equipotent to those of PGI2 released by ANG II, estimated from dose response curves for TRK-100 and enhancement by indomethacin of ANG II-induced contractions, did not differ in proximal and distal portions. It appears that differences in the contractile response to ANG II depend on heterogeneity in ANG II receptors responsible for the arterial contraction, but not on differences in the PGI2 generation via ANG II receptor activation. PMID- 3048223 TI - Dural arteriovenous malformation of the tentorium. Report of a case and a review of the literature. PMID- 3048222 TI - The protective effect of glucose, but not 3-O-methyl glucose, against alloxan induced diabetes depends upon the route of hexose administration. AB - It is well established that pretreatment with glucose prevents the development of alloxan-induced diabetes in rodents. This was confirmed in the present study, and a protective effect against the development of hyperglycemia, after alloxan administration (75 mg/kg body weight), in male NMRI mice was observed when glucose was injected i.v. 10 or 20 min before alloxan injection. However, when glucose was administered i.p., a protective effect was only observed when glucose was given 20 min before alloxan, despite the fact that the serum glucose concentrations at the time of the alloxan injection were similar in the i.v.- and i.p.-injected animals. Administration of 3-O-methyl glucose by i.v. and i.p. routes protected against the effects of alloxan to the same extent. When alloxan was administered at a lower dose (65 mg/kg body weight) both i.v. and i.p. administration of glucose 10 min before alloxan, protected against the development of hyperglycemia. It is concluded that the route of administration of glucose, but not of 3-O-methyl glucose, affects its protective action against alloxan-induced diabetes. The present findings suggest that an important feature of the glucose mediated protection against alloxan-induced diabetes is the metabolism of glucose within the B-cells, rather than via extracellular effects on the B-cell plasma membrane. PMID- 3048224 TI - Multiple-dose arecoline infusions in Alzheimer's disease. AB - Twelve patients with dementia of the Alzheimer type received two-hour infusions of placebo and the muscarinic cholinergic agonist arecoline hydrobromide at rates of 1, 2, and 4 mg/h in a double-blind, randomized fashion. These infusions resulted in dose-dependent physiologic and neuroendocrine effects consistent with central cholinergic stimulation. Infusions were generally well tolerated. No statistically significant improvement in performance on most cognitive tasks assessing knowledge memory and episodic learning and memory was observed at any dose, although marginal improvement in picture recognition ability and in ratings of word-finding were observed at the lower doses. Psychomotor activation and slightly improved affect were reliably observed at the lower doses, whereas increasing psychomotor retardation was observed at the highest dose. The data support a role for central cholinergic modulation of some aspects of cognition, behavior, and affect in this population. The apparent greater behavioral sensitivity of patients with Alzheimer's disease in comparison with subject populations previously studied, as well as the altered dose responsiveness, merit further study in relationship to normal aging. PMID- 3048225 TI - The effects of acute scopolamine in geriatric depression. AB - In an intensive multidrug, multidose study, nine elderly depressed patients were administered 0.1, 0.25, and 0.5 mg of scopolamine hydrobromide, 1 mg of oral lorazepam, and placebo in a double-blind investigation aimed at assessing the status of the central cholinergic nervous system in geriatric depression. Significant cognitive and behavioral effects of scopolamine were observed only at the high dose (0.5 mg), while lower doses and lorazepam showed no significant differences from placebo. Cognitive deficits caused by scopolamine were in the areas of new learning, access to semantic memory, vigilance, and continuous performance. Behavioral effects consisted of activation, restlessness, and anxiety, but there was no significant effect on depressed mood. These results suggest that elderly depressed patients with mild to moderate cognitive impairment seem to be more similar to previously studied elderly controls rather than to patients with Alzheimer's disease in their reaction to short-term cholinergic blockade, and suggest that the cognitive and mood changes often seen in geriatric depression may involve factors other than disturbed muscarinic cholinergic mechanisms. PMID- 3048227 TI - Autism and genetics. A decade of research. AB - The last ten years of research on the genetics of infantile autism were critically reviewed. Epidemiologic findings have shown that autism is a rare disorder with a prevalence of two to five per 10,000, a male-female ratio of 3:1, and an association with mental retardation (66% to 75% of autistic subjects have full-scale IQ scores [70]). Autism is familial, as reflected in an empiric sibling recurrence risk of 3% and pooled monozygotic and dizygotic concordance rates of 64% and 9%, respectively, which are much greater than the population prevalence of 0.02% to 0.05%. Genetic heterogeneity is pronounced with potential genetic subgroups, including autosomal recessive inheritance, X-linked inheritance, and sporadic chromosomal anomalies. Studies of subclinical markers in autism have elucidated potential markers at various levels of phenotypic expression from the DNA to the behavioral level. Linkage and cytogenetic studies point to two chromosome regions as putative markers, 9q34 and Xq27. Results of family studies support a putative biochemical marker, low levels of plasma dopamine-beta-hydroxylase, and a putative cognitive marker, ie, normal visuospatial but low verbal functioning, in autism. The frequency of minor physical anomalies and presence or absence of mental retardation are two dimensions of the physical and behavioral phenotype that may demark etiologically distinct subgroups. Genetic heterogeneity is offered as one explanation of the observed sex difference in the prevalence of autism. Directions for potentially fruitful research should be considered. PMID- 3048226 TI - Social zeitgebers and biological rhythms. A unified approach to understanding the etiology of depression. AB - Results of biological and psychosocial studies of depression completed in the last decade have stimulated the need for new hypotheses that synthesize these findings in a unified etiologic theory. The importance of disruption of biological rhythms on the one hand, and psychosocial losses on the other, in the causation of depressive episodes suggest one possible unifying hypothesis. The concept of loss of "social zeitgebers," ie, persons, social demands, or tasks that set the biological clock, may provide the link between biological and psychosocial theories of etiology. We suggest that a disruption of social rhythms, which may result in instability in biological rhythms, could be responsible for triggering the onset of a major depressive episode in vulnerable individuals. PMID- 3048229 TI - Depression and suicide in adolescence. PMID- 3048228 TI - Immunoglobulins in human dental caries. AB - The penetration and complement fixation of sIgA, IgM and IgG were studied in advanced human dental caries with double-staining immunofluorescence technique. Immunoglobulins were found in dental plaque and in the superficial layers of the lesions which were exposed to saliva. sIgA was the most frequent, followed by IgG and IgM. IgG had the greatest capacity for penetration into the lesions, followed by IgM and sIgA. IgM was most frequently found in association with complement C3. PMID- 3048230 TI - Eating disorders in the adolescent. PMID- 3048231 TI - Fetal alcohol syndrome. PMID- 3048232 TI - Experimental study on facial nerve suturing--comparison between epineural and perineural sutures. AB - A comparison was made between epineural and perineural sutures using the orbicular is oculi branches of facial nerves of 30 cats. Nerve regeneration was slightly reduced with perineural suture without a tube compared with epineural suture with a tube, epineural suture without a tube and perineural suture with a tube, which showed similar results. A good regeneration was observed when approximation was made without tension at both cut ends of the perineurium. When the corresponding funiculus cannot be identified, epineural suture is considered sufficient. PMID- 3048233 TI - Explaining coma arousal therapy. PMID- 3048234 TI - Diagnosis of cystic fibrosis. PMID- 3048236 TI - Urological anomalies detected on antenatal ultrasound: a 9 year review. AB - One hundred and forty-eight patients (107 male, 41 female), in whom a urological anomaly was detected on antenatal ultrasound examination, are reviewed. Postnatal imaging was done primarily by ultrasonography (US) which was often repeated. Depending upon the ultrasound findings, the patients had a renal nuclide scan (RNS) and/or micturating cysto-urethrogram (MCU), but intravenous urogram (IVU) was not usually considered necessary. A range of urological anomalies was encountered, but renal anomalies were most common. Over half the cases had anomalies which did not require surgery, with non-obstructive pelvicalyceal dilatation being frequent. Almost half the operated cases had features which should have allowed a clinical diagnosis without the knowledge of the antenatal findings. A fifth of the cases were occult in that they would not have been diagnosed early in life but for the antenatal detection. The majority had congenital pelviureteric junction obstruction and results of early reconstructive surgery were satisfactory. PMID- 3048235 TI - Use of methylprednisolone as prophylaxis for immediate adverse infusion reactions in hypogammaglobulinaemic patients receiving intravenous immunoglobulin: a controlled trial. AB - Ten children and young adults with humoral immunodeficiency disorders were enrolled in a blind crossover study of the effect of intravenous methylprednisolone in preventing immediate adverse infusion reactions to intravenous immunoglobulin. The patients selected had all experienced frequent adverse reactions previously. Each patient received two infusions of intravenous immunoglobulin at least 4 weeks apart. Infusions were of constant volume and rate for each patient. Intravenous methylprednisolone in a dose of 1 mg/kg bodyweight was given 20 min prior to one of the infusions only. A significant decrease in the severity of infusion-associated immediate adverse reactions was seen following the administration of methylprednisolone (P less than 0.01). Interruption of the infusion was necessary for only one patient when methylprednisolone prophylaxis was given, but temporary cessation of the infusion was required for eight of the 10 patients when methylprednisolone was not given (P less than 0.05). PMID- 3048237 TI - Systemic therapy with aminoglycoside antibiotics in the horse. PMID- 3048238 TI - Comparison of media used for the primary isolation of Mycobacterium bovis by veterinary and medical diagnostic laboratories. AB - Veterinary and medical laboratories engaged in the cultural diagnosis of bovine or human tuberculosis were requested to supply samples of the media that they routinely use for the primary isolation of M. bovis. Fourteen laboratories supplied 7 basic media types; these were Lowenstein-Jensen, Stonebrink's, modified Middlebrook 7H11 agar, tuberculosis bovine blood agar, egg yolk agar, Gerloff's egg and Herrold's egg yolk. Two strains of M. bovis were used to test the media, strain AN5, a glycerol-tolerant laboratory strain and M86/90 a glycerol-sensitive wildtype strain. AN5 grew well on all media with the exception of Herrold's and strain M86/90 did not grow on media containing glycerol and grew poorly on Herrold's medium. It is recommended that Lowenstein-Jensen with pyruvate (but without glycerol), Stonebrink's, modified Middlebrook 7H11 and tuberculosis bovine blood agar should be considered the media of choice for the primary isolation of M. bovis. Egg yolk agar also proved adequate for this purpose in the trial. This medium may be suitable for routine use but to date experience with its use is limited. PMID- 3048241 TI - [Pathogenesis and immunoprophylaxis of Pasteurella haemolytica infections (review)]. PMID- 3048239 TI - Some aspects of the epidemiology of equine salmonellosis. AB - A survey of 2 horse populations was done to detect the number of asymptomatic faecal excretors of Salmonella sp. 1201 faecal samples from 250 horses hospitalised at the University of Sydney were cultured. Three serotypes, S. typhimurium (4 horses), S. anatum (2) and S. tennessee (1) were isolated from 7 horses (2.8%). None was detected in 75 mares similarly examined at a thoroughbred stud farm. In retrospect, S. typhimurium was also the most common (70%) of the 19 serotypes recovered from 171 horses with clinical salmonellosis seen at Camden, 1969 to 1986. Forty cases occurring since 1983 were reviewed in detail; the mortality rate was high (60%) and an increased proportion was due to S. bovis morbificans. Five horses developed salmonellosis while hospitalised and it was usually impossible to be certain whether these cases developed from the carrier state into overt disease or resulted from infections acquired in hospital. PMID- 3048240 TI - Tooth eruption: an emerging enigma. PMID- 3048242 TI - [Fertility control of dairy cows in the pre-service and conception period]. PMID- 3048243 TI - [Detection of thermolabile enterotoxins of Escherichia coli with the Vero cell test, ganglioside (GM1)-enzyme-linked immunosorbent assay and latex agglutination test--a comparative study]. PMID- 3048244 TI - Kinetic mechanism of pulmonary carbonyl reductase. AB - The kinetic mechanism of guinea-pig lung carbonyl reductase was studied at pH 7 in the forward reaction with five carbonyl substrates and NAD(P)H and in the reverse reaction with propan-2-ol and NAD(P)+. In each case the enzyme mechanism was sequential, and product-inhibition studies were consistent with a di-iso ordered bi bi mechanism, in which NAD(P)H binds to the enzyme first and NAD(P)+ leaves last and the binding of cofactor induces isomerization. The kinetic and binding studies of the cofactors and several inhibitors such as pyrazole, benzoic acid, Cibacron Blue and benzamide indicate that the cofactor and Cibacron Blue bind to the free enzyme whereas the other inhibitors bind to the binary and/or ternary complexes. PMID- 3048245 TI - Nucleotide sequence analysis and overexpression of the gene encoding a type III chloramphenicol acetyltransferase. AB - The gene catIII, encoding a type III enterobacterial chloramphenicol acetyltransferase, was cloned from the transmissible plasmid R387 into pBR322 and bacteriophage M13 mp8. Nucleotide sequence analysis of 1160 bp of DNA identified an open reading frame encoding a protein of 213 amino acid residues and a calculated molecular mass of 24965 Da. The predicted N-terminal sequence is identical with that determined by Edman degradation of chloramphenicol acetyltransferase purified from Escherichia coli harbouring R387. Sequences equivalent to the consensus motifs for initiation and rho-factor-independent termination of transcription in E. coli occur 5' and 3' to the catIII open reading frame. In contrast with the catI gene, present on transposon Tn9 and many enterobacterial plasmids, expression of catIII is not subject to cyclic AMP mediated catabolite repression in vivo and there is no sequence in the 5' non coding DNA that resembles that deduced as the consensus for the binding of cyclic AMP receptor protein. Unique restriction-endonuclease cleavage sites were introduced adjacent to the catIII reading frame by using oligonucleotide-directed mutagenesis to facilitate insertion into E. coli expression vectors. Fully active chloramphenicol acetyltransferase represents 30-50% of the soluble protein component of cell-free extracts of E. coli containing the appropriate plasmids. PMID- 3048246 TI - Characterization of the lipid acyl hydrolase activity of the major potato (Solanum tuberosum) tuber protein, patatin, by cloning and abundant expression in a baculovirus vector. AB - Patatin is a family of glycoproteins that accounts for 30-40% of the total soluble protein in potato (Solanum tuberosum) tubers. This protein has been reported not only to serve as a storage protein, but also to exhibit enzymic activity. By using a baculovirus system to express protein from the patatin cDNA clone pGM01, it was unambiguously shown that the patatin coded by this DNA has lipid acyl hydrolase and acyltransferase activities. The enzyme is active with phospholipids, monoacylglycerols and p-nitrophenyl esters, moderately active with galactolipids, but is apparently inactive with di- and tri-acylglycerols. PMID- 3048248 TI - Effect of specific proteolytic cleavages on tubulin polymer formation. AB - The capacity for self-polymerization and shape of the tubulin polymers assembled after digestion with trypsin, Pronase, chymotrypsin, subtilisin, Staphylococcus aureus proteinase V8 and proteinase K were investigated. Digestion with trypsin, Pronase or chymotrypsin resulted in a decrease in the ability of tubulin for self assembly, whereas limited proteolysis with subtilisin, S. aureus proteinase V8 or proteinase K resulted in an increase in such ability. The shape of the assembled polymers varied from typical microtubules (after the treatment with trypsin or Pronase) to sheets (after the treatment with chymotrypsin) and from hooked microtubules with a constant polarity (after the treatment with subtilisin) to the disappearance of a defined polarity of such polymers (after the treatment with S. aureus V8 proteinase or proteinase K). These results indicate that the tubulin C-terminal regions are involved in the regulation of microtubule polymerization, shape, directional growth and lateral interactions between tubulin protofilaments. PMID- 3048247 TI - Insulin and insulin-like growth factor 1 stimulate the phosphorylation on tyrosine of a 160 kDa cytosolic protein in 3T3-L1 adipocytes. AB - Insulin and IGF-1 (insulin-like growth factor 1) rapidly stimulate the phosphorylation on tyrosine of a 160 kDa cytosolic protein (pp160) in intact 3T3 L1 adipocytes. Half-maximal phosphorylation of pp160 is attained with either 4 nM insulin or 20 nM-IGF-1. A semi-quantitative immunoblotting procedure using anti phosphotyrosine antibody revealed that the insulin-stimulated 3T3-L1 adipocyte possesses approx. 3 x 10(5) and 0.6 x 10(5) phosphotyrosyl sites, respectively, in pp160 and insulin receptor beta-subunit. Removal of insulin from stimulated cells results in the rapid (within 15 min) loss of phosphate groups from tyrosyl residues in both pp160 and receptor beta-subunit. Whereas pp160 remains maximally phosphorylated on tyrosine for up to 60 min in the presence of 100 nM-insulin, IGF-1 at the same concentration induces only a transient response that is maximally 50% of that observed with insulin. pp160 is not phosphorylated on tyrosine in response to platelet-derived growth factor or epidermal growth factor. Although pp160 appears to be a soluble cytoplasmic protein, in the presence of 1 mM-ZnCl2 it becomes membrane-associated. In view of its apparent cytoplasmic localization and its inability to bind to either wheat-germ agglutinin or concanavalin A, pp160 does not appear to be a typical glycoprotein growth-factor receptor. Our results suggest that pp160 may be a physiologically important cellular substrate of the insulin-receptor tyrosine kinase in the intact 3T3-L1 adipocyte. PMID- 3048249 TI - Insulin-induced myosin light-chain phosphorylation during receptor capping in IM 9 human B-lymphoblasts. AB - We have examined further the interaction between insulin surface receptors and the cytoskeleton of IM-9 human lymphoblasts. Using immunocytochemical techniques, we determined that actin, myosin, calmodulin and myosin light-chain kinase (MLCK) are all accumulated directly underneath insulin-receptor caps. In addition, we have now established that the concentration of intracellular Ca2+ (as measured by fura-2 fluorescence) increases just before insulin-induced receptor capping. Most importantly, we found that the binding of insulin to its receptor induces phosphorylation of myosin light chain in vivo. Furthermore, a number of drugs known to abolish the activation properties of calmodulin, such as trifluoperazine (TFP) or W-7, strongly inhibit insulin-receptor capping and myosin light-chain phosphorylation. These data imply that an actomyosin cytoskeletal contraction, regulated by Ca2+/calmodulin and MLCK, is involved in insulin-receptor capping. Biochemical analysis in vitro has revealed that IM-9 insulin receptors are physically associated with actin and myosin; and most interestingly, the binding of insulin-receptor/cytoskeletal complex significantly enhances the phosphorylation of the 20 kDa myosin light chain. This insulin-induced phosphorylation is inhibited by calmodulin antagonists (e.g. TFP and W-7), suggesting that the phosphorylation is catalysed by MLCK. Together, these results strongly suggest that MLCK-mediated myosin light-chain phosphorylation plays an important role in regulating the membrane-associated actomyosin contraction required for the collection of insulin receptors into caps. PMID- 3048250 TI - Escherichia coli promoter -10 and -35 region homologies correlate with binding and isomerization kinetics. AB - The DNA promoter sequence at which gene transcription is initiated in Escherichia coli contains two distinct regions of conserved nucleotides. Coefficients of homology to the -10 region and the -35 region were computed for many different prokaryotic promoters. Linear equations were derived that relate the degree of homology for each promoter region to the two kinetic constants that describe the interaction of RNA polymerase with the promoter site on DNA: the strength KB of binding to form closed complex, and the rate kf of isomerization to form open complex. A graphical plot of -35 versus -10 promoter region homologies for many promoters suggest that certain classes of prokaryotic operons might utilize differential degrees of consensus homology in these two regions to achieve specific control over kf and KB, in addition to modulating overall promoter strength. PMID- 3048252 TI - The effects in vivo of mutationally modified uroporphyrinogen decarboxylase in different hem12 mutants of baker's yeast (Saccharomyces cerevisiae). AB - Nine new hem12 haploid mutants of baker's yeast (Saccharomyces cerevisiae), totally or partially deficient in uroporphyrinogen decarboxylase activity, were subjected to both genetic and biochemical analysis. The mutations sites studied are situated far apart within the HEM12 gene located on chromosome IV. Uroporphyrinogen decarboxylase activity in the cell-free extracts of the mutants was decreased by 50-100%. This correlated well with the decrease of haem formation and the increased accumulation and excretion of porphyrins observed in vivo. The pattern of porphyrins (uroporphyrin and its decarboxylation products) accumulated in the cells of mutants partially deficient in uroporphyrinogen decarboxylase activity did not differ significantly, although differences in vitro were found in the relative activity of the mutant enzyme at the four decarboxylation steps. The excreted porphyrins comprised mainly dehydroisocoproporphyrin or pentacarboxyporphyrin. In heterozygous hem12-1/HEM12 diploid cells, a 50% decrease in decarboxylase activity led to an increased accumulation of porphyrins as compared with the wild-type HEM12/HEM12 diploid, which points to the semi-dominant character of the hem12-1 mutation. The biochemical phenotypes of both the haploid and the heterozygous diploid resembles closely the situation encountered in porphyria cutanea tarda, the most common human form of porphyria. PMID- 3048253 TI - Fatty acid metabolism in hepatocytes cultured with hypolipidaemic drugs. Role of carnitine. AB - The direct effects of clofibrate analogues on carnitine acyltransferase activities and fatty acid metabolism were studied in cultured hepatocytes. Rat hepatocytes cultured with bezafibrate or ciprofibrate (0.1-10 micrograms/ml) for 48 h had increased activities of carnitine acetyltransferase (CAT; 4-6-fold) and carnitine palmitoyltransferase (CPT; 12-34%). The increase in CAT was higher in hepatocytes from the periportal zone (440%) of rat liver compared with cells from the perivenous zone (266%). In human hepatocytes, in contrast with rat, the fibrates did not cause a marked increase in CAT activity. The effects of fibrates on palmitate metabolism were dependent on the carnitine status. In the presence of exogenous carnitine (1 mM), rat hepatocytes cultured with bezafibrate had higher rates of total palmitate metabolism (29-34%) without increased partitioning of palmitate towards beta-oxidation, relative to control cultures. At low endogenous carnitine concentrations, cells cultured with bezafibrate had a greater increase in palmitate metabolism, esterification and cellular accumulation of triacylglycerol compared with the corresponding increases in the presence of carnitine. The changes in palmitate metabolism at either high or low carnitine concentrations were small in comparison with the changes in CAT activity. It is concluded that the increase in hepatic carnitine that occurs in vivo after fibrate feeding probably plays the major role in the changes in partitioning of fatty acid between beta-oxidation and esterification. PMID- 3048255 TI - Progesterone and prolactin are both required for suppression of the induction of rat alpha-lactalbumin activity. AB - Progesterone prevents lactation during pregnancy. This anti-lactogenic effect includes suppression of the advent of alpha-lactalbumin activity, an effect which prevents the formation of lactose. Alpha lactalbumin activity can be induced to some extent in pregnant rat mammary explants by insulin and hydrocortisone alone, and to a greater extent with prolactin in addition, or with EGF in addition. Physiological levels of progesterone markedly inhibit the induction in the presence of prolactin plus insulin and hydrocortisone, only weakly inhibit in the presence of insulin and hydrocortisone alone, and have no inhibitory effect in the presence of EGF plus insulin and hydrocortisone. Prolactin permits some inhibition in the presence of EGF. The results suggest that progesterone does not subvert the essential insulin or glucocorticoid signals. It also appears that transduction of the prolactin signal is required in order that progesterone effectively block induction of alpha-lactalbumin activity. PMID- 3048256 TI - Hemostasis in larvae of Manduca sexta: formation of a fibrous coagulum by hemolymph proteins. AB - Little is known of the participation of insect hemolymph proteins in wound healing and clot formation. We describe the assembly of purified hemolymph protein from the tobacco hornworm into an extended fibrous coagulum in the absence of hemocytes. This coagulum resembles the clot formed from bovine fibrinogen and thrombin. Structural components of the coagulum are present in hemolymph, however, spontaneous assembly occurs only in hemolymph collected through a wound. The fibrous coagulum assembles from purified structural protein(s) following addition of a non-protein factor from hemolymph, which is also present in Grace's insect cell culture medium. PMID- 3048254 TI - Effects of hyperthyroidism on the sensitivity of glycolysis and glycogen synthesis to insulin in the soleus muscle of the rat. AB - 1. The effects of hyperthyroidism on the sensitivity and responsiveness of glycolysis and glycogen synthesis to insulin were investigated in the isolated incubated soleus muscle of the rat. 2. Hyperthyroidism, which was induced by administration of tri-iodothyronine (T3) to rats for 2, 5 or 10 days, increased fasting plasma concentrations of glucose, insulin and free fatty acids. 3. Administration of T3 for 2 or 5 days increased the rates of glycolysis at all insulin concentrations studied: this was due to increased rates of both glucose phosphorylation and glycogen breakdown, but there was no effect of T3 on the sensitivity of glycolysis to insulin. However, administration of T3 for 10 days increased the sensitivity of the rate of glycolysis to insulin. 4. The concentration of adenosine in the gastrocnemius muscles of the rats was not different from controls after 5 days, but it was markedly decreased after 10 days of T3 administration. If these changes are indicative of changes in the soleus muscle, the increased sensitivity of glycolysis to insulin found after 10 days' T3 administration could be due to the decrease in the concentration of adenosine. 5. Administration of T3 decreased the sensitivity of glycogen synthesis to insulin and the glycogen content of the soleus muscles. This may explain the decreased rates of non-oxidative glucose disposal found in spontaneous and experimental hyperthyroidism in man. 6. The rates of glucose oxidation did not change after 2 days, but they were increased after 5 and 10 days of T3 administration. PMID- 3048257 TI - Acute-phase response of mRNAs for serum amyloid P component, C-reactive protein and prealbumin (transthyretin) in mouse liver. AB - Acute-phase response of mRNAs for serum amyloid P component (SAP), C-reactive protein (CRP) and prealbumin was examined in C57BL/6 mouse liver by hybridization to specific cDNA probes. Although the level of SAP mRNAs in the unstimulated mouse was about one-tenth of that of CRP mRNAs, it increased up to 60-fold during the first 20 hr, and returned gradually to the original level at 69 hr after the administration of Escherichia coli lipopolysaccharide. On the other hand, the level of CRP mRNA rapidly increased up to 6-fold during the first 4 hr, and reverted to the original level as early as at 20 hr. In contrast, the level of mRNA for prealbumin decreased to about 0.5-fold during the first 20 hr, recovered and increased up to 1.6-fold of the original level during 32 to 69 hr. PMID- 3048258 TI - The effect of platelet activating factor on electron transport of spinach chloroplasts. AB - It was found that platelet activating factor (1-O-alkyl-2-acetyl-sn-glycero-3 phosphocholine) inhibits electron transport greater than 90% in Photosystem II of spinach chloroplasts in concentrations from 2.8 to 3.5 micrograms/ml. The inhibition of the main pathway of electron transport through Photosystem II appeared to be specific for the platelet activating factor. Phorbol myristate acetate, 1,2-dipalmitin or fatty acid esters gave 17-32% inhibition in higher concentrations. The inhibition site for platelet activating factor was localized close to the reaction center of Photosystem II, based on the inhibition of the donor reaction, diphenyl carbazide----indophenol, in Tris-treated chloroplasts. Other Photosystem II reactions, H2O----silicomolybdic acid, H2O----2,5 dimethylbenzoquinone, or H2O----indophenol, were also inhibited by platelet activating factor. The present data point out the unique inhibition of Photosystem II electron transport by the platelet activating factor, but do not support the operation of a phosphoinositide cycle in chloroplasts. PMID- 3048259 TI - Amyloid fibrils formed from a segment of the pancreatic islet amyloid protein. AB - A synthetic peptide formed from residues 20-29 of the pancreatic islet amyloid protein has the confirmation of a twisted beta-pleated sheet protein suggesting it is a potential contributor toward amyloid fibril formation in the islets of Langerhans in Type 2 diabetes mellitus. PMID- 3048260 TI - Characterization of the glucose transporter from rat brain synaptosomes. AB - Our goal was to characterize the glucose transporter in synaptosomes and to compare it to the different forms of transporter already identified. Cross reactivity with antibodies to the human erythrocyte transporter, Km of glucose uptake, reversibility of NEM inhibition of transport, and insulin sensitivity were all examined. Immunoblotting showed a band at Mr 40,000, and the Km of glucose uptake was determined to be about 4 mM. Treatment with NEM caused irreversible inhibition of glucose uptake, while incubation with insulin failed to stimulate uptake. The results suggest that the transporter in synaptosomes resembles the human erythrocyte transporter. PMID- 3048261 TI - Evidence for a new ribonucleotide reductase in anaerobic E. coli. AB - E. coli conditional iron-containing ribonucleotide reductase (Fe-RR) mutant and wild type strains grew anaerobically under conditions when Fe-RR was absent or inhibited. Furthermore, a B12-independent, hydroxyurea-resistant RR activity, unaffected by monoclonal antibodies against either subunit B1 or B2 of Fe-RR, was partially purified from anaerobically grown mutant and wild-type E. coli. These findings indicate that E. coli has a second RR representative of a new class of RRs and that this is the first report where both in vivo and in vitro evidence is presented. It is probable that other facultative anaerobes also have two different RRs such that an optimal supply of deoxyribonucleotides is maintained under all growth conditions. PMID- 3048262 TI - Cell membrane, but not circulating, carcinoembryonic antigen is linked to a phosphatidylinositol-containing hydrophobic domain. AB - Carcinoembryonic antigen is present in the cell membrane of most tumors of colorectal origin and in the plasma of patients with colorectal cancer and other malignancies. In this paper we demonstrate that carcinoembryonic antigen can be released from HT-29 cells by phosphatidylinositol specific phospholipase C. Triton X-114 phase separation shows that phospholipase C converts the antigen into a water soluble protein. In addition, plasma carcinoembryonic antigen behaves as the cleaved antigen in phase separation experiments. This strongly suggests that carcinoembryonic antigen is attached to cell membranes by a glycosyl-phosphatidylinositol anchor and that it can be released in vivo by enzymatic cleavage of the hydrophobic tail. PMID- 3048263 TI - Secretion of corticotropin releasing factor (CRF) and vasopressin (AVP) into the hypophysial portal blood of conscious, unrestrained rams. AB - The secretion of Corticotropin-Releasing Factor (CRF) and Arginine Vasopressin (AVP) into sheep hypothalamo-hypophysial portal blood was investigated in conscious, unrestrained castrated rams. One to two hours after the onset of the collection, peripheral ACTH and cortisol levels were within the basal range in all the animals and portal CRF and AVP concentrations were low ranging between 5 and 90 pg/ml. After acute hemorrhage, a sharp increase in portal CRF and AVP was observed, inducing a sustained elevation of peripheral ACTH and cortisol. These data show that measurements of CRF and AVP in the portal blood of conscious sheep provide a good methodology to study the regulation of basal and stress-induced ACTH secretion. PMID- 3048265 TI - The influence of insulin and glucagon on the interactions between glycolytic enzymes and cellular structure. AB - The influence of insulin and glucagon on the release of glycolytic enzyme activities and actin from cultured pig kidney cells treated with digitonin has been studied. Both insulin and glucagon reduced the release of all glycolytic enzymes except for phosphofructokinase, and concurrently reduced the release of actin. These data have been discussed in relation to their contribution to knowledge of the interactions between glycolytic enzymes and actin filaments of the cytoskeleton, and to the influence of hormones on these interactions. PMID- 3048264 TI - Cellular localization of insulin-degrading enzyme in rat liver using monoclonal antibodies specific for this enzyme. AB - Although insulin-degrading enzyme (IDE) has been implicated in the intracellular degradation of insulin, the cellular localization of this enzyme is still controversial. In the present study, we have examined the cellular localization of IDE in the rat liver by three different techniques using monoclonal antibodies. First, direct immunohistochemical staining of rat liver with one of the monoclonal antibodies revealed that IDE immunoreactivity mainly exists in parenchymal cells, especially in the vicinity of the portal tract and also in the epithelium of the bile duct under light microscopy. In the electron microscopic study, IDE immunoreactivity was found in the cytoplasm near the rough endoplasmic reticulum but not in the plasma membrane, nucleus, or mitochondria. Second, immunoblotting analysis of the subcellular fraction in rat liver showed that the monoclonal antibody specifically reacted with a single polypeptide in the cytosolic fraction, of apparent Mr 110,000, which was consistent with the Mr of IDE. However, a polypeptide band corresponding to IDE could not be observed in the plasma membrane, mitochondrial, or lysosomal fraction. Third, IDE was only detectable in the cytosolic fraction by sandwich radioimmunoassay using two monoclonal antibodies. These results all suggest that IDE is a cytosolic enzyme. PMID- 3048266 TI - A two-site solid phase enzyme immunoassay for rat epidermal growth factor. AB - A sensitive and simple two-site enzyme immunoassay was developed for rat epidermal growth factor (rEGF), which was purified from male rat submaxillary glands. This system is based on the sandwiching of the antigen between anti-rat EGF IgG antibody coated on a polystyrene bead and anti-rat EGF Fab' antibody linked peroxidase, whose activity was stable at 4 degrees C for 6 months. In this system, the rat EGF was detectable at a concentration of 2 pg/tube. No interference was observed by addition of human plasma. There was no cross reactivity with mouse EGF, human EGF, or other substances with similar chemical structures. PMID- 3048251 TI - Surface proteins and glycoproteins of human leucocytes. PMID- 3048267 TI - Participation of histidine in biosynthesis of the pyrimidine moiety of thiamin in Saccharomyces cerevisiae. AB - The amide nitrogen atom of glutamine is incorporated into the pyrimidine moiety of thiamin in Escherichia coli and Saccharomyces cerevisiae. However, addition of casamino acids to the medium increases incorporation of the amide nitrogen atom of glutamine in E. coli, but decreases it in S. cerevisiae. This suggests that some amino acids other than glutamine in casamino acids are more direct precursors of the pyrimidine moiety in S. cerevisiae. To determine the direct precursor, we investigated the competitive effect of 14N-amino acids on the incorporation of 15NH4Cl into the pyrimidine moiety and found that histidine decreased the incorporation of 15N. Thus, histidine was concluded to be the direct precursor of the nitrogen atom of the pyrimidine moiety of thiamin in S. cerevisiae. PMID- 3048268 TI - Cytochrome P-450 alterations in the BB/Wor spontaneously diabetic rat. AB - The hepatic content of two individual cytochrome P-450 enzymes was analyzed in spontaneously diabetic (BB/Wor) male rats. The major male-specific form, RLM5, was found to be slightly decreased in livers of male rats shortly after the onset of diabetes. In contrast, the level of RLM6 was elevated in livers of diabetic rats that had not received insulin and had become ketotic. These results confirm that the changes in some individual forms of cytochrome P-450 after chemical (streptozotocin) induction of diabetes are also seen in the spontaneously diabetic animal. The earlier observed alterations in cytochrome P-450 are therefore due to the state of diabetes and not to inductive or depressive effects of streptozotocin. PMID- 3048269 TI - [Synthesis of lipid A analogs: achievements and perspective]. AB - Data on the synthesis of analogues of lipid A, a biologically active fragment of gram-negative bacteria's lipopolysaccharides, are summarized. Main types of the compounds obtained are systematized, and problems of the synthesis of various parts of the molecule considered. The results of studying biological activity of lipid A analogues are discussed, which led to some conclusions on the structure function relation. Perspectives of further studies are briefly outlived. PMID- 3048270 TI - [Affinity modification of Escherichia coli ribosomes with the non-hydrolyzed GTP analog, gamma-[4-N-(2-chloroethyl)-N-methylaminobenzyl]amide of GTP]. AB - Substituted gamma-amides of GTP viz. GTP gamma-[4-N-(2-chloro- and gamma-[4-N-(2 hydroxyethyl)-N-methylaminobenzyl]amide (CIRCH2NHpppG and OHRCH2NHpppG, resp.) were shown to be unhydrolisable GTP analogues in the EF-Tu-dependent GTP-ase reaction of ribosomes. The reactive analogue, CIRCH2NHpppG, was used for affinity labelling within the 70S ribosome.poly(U).tRNAPhe(P-site).Phe-tRNAPhe.EF Tu.CIR[14C]CH2.NHpppG complex. Both 50S and 30S subunits were thus labelled but 50S subunit was modified considerably more than 30C subunit. Labelled were proteins L17, L21, S16, S21, and rRNA of both subunits, 23C rRNA within 50C subunit being labelled preferentially as compared with 50C proteins. No labelling of EF-Tu within the complex was detected. PMID- 3048271 TI - [Genes encoding bacterial RNA-polymerase. I. Cloning of rpoBC-operon of Pseudomonas putida and its physical map]. AB - The P. putida rpoBC operon, coding for beta and beta' subunits of RNA polymerase, was cloned and its physical map constructed. PMID- 3048273 TI - A controlled study of the effects of a supervised cardiovascular fitness training program on the manifestations of primary fibromyalgia. AB - Forty-two patients with primary fibromyalgia were randomized into a 20-week program consisting of either cardiovascular fitness (CVR) training or simple flexibility exercises (FLEX) that did not lead to enhanced cardiovascular fitness. Patients were supervised by the same medical fitness instructors. Patients in neither group had contact with members of the other group, and were blinded as to the exercise taught to the alternative group. Groups met for 60 minutes 3 times each week. The compliance rate was 90%. Thirty-eight patients completed the study (18 with CVR training and 20 with FLEX). Blind assessments (standardized in preliminary trials to achieve acceptable inter-rater agreement) were performed by the same 2 examiners. After 20 weeks, patients receiving CVR training showed significantly improved cardiovascular fitness scores compared with those receiving FLEX training (t[35] = -4.22, P less than 0.003). Logistic regression analysis showed clinically and statistically significant improvements in pain threshold scores, which were measured directly over fibrositic tender points, in patients undergoing CVR (t[35] = 2.21, P less than 0.04). There was also a trend toward improvement in pain scores (visual analog scale) in the CVR group, but this did not reach statistical significance. There was no improvement in the percentage of body area affected by fibrositic symptoms or the number of nights per week or hours per night of disturbed sleep (self-report inventories). However, compared with the FLEX group, the CVR-trained patients improved significantly in both patient and physician global assessment scores.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3048272 TI - Studies of a common idiotype PR4 in autoimmune rheumatic disease. AB - A new common idiotype, designated PR4, is described. This idiotype was originally identified on a human hybridoma-derived monoclonal antibody from a patient with leprosy, which binds the major Mycobacterium leprae-derived antigen, phenolic glycolipid-1, poly(ADP)-ribose, DNA, and poly(dT). The PR4 idiotype was found in patients with systemic lupus erythematosus (SLE) (70%), rheumatoid arthritis (40%), and Sjogren's syndrome (15%). It was not, however, found in the spouses of the SLE patients or (unlike other lupus idiotypes) in their healthy first-degree relatives. Although no correlation between PR4 idiotype levels and disease activity in SLE was found, a subset of rheumatoid arthritis patients with high levels of the idiotype was identified. PMID- 3048274 TI - An evaluation of serum osteocalcin in Paget's disease of bone and its response to diphosphonate treatment. AB - We found that serum bone gamma-carboxyglutamic acid-containing protein (BGP) (osteocalcin) had lower sensitivity and specificity for measurement of disease activity in Paget's disease of bone than other biochemical measures of disease activity. The administration of diphosphonates induced suppression of urinary hydroxyproline excretion and a subsequent decrease in alkaline phosphatase values, but no consistent change in BGP values. Serum BGP measurements have limited value as a screening test for Paget's disease or for monitoring treatment of the disorder. PMID- 3048275 TI - Mechanisms of joint damage in an experimental model of hemophilic arthritis. AB - Autologous whole blood was injected into the knee joints of rabbits 3 times each week for 12 weeks. The resulting destructive arthritis showed macroscopic and microscopic changes similar to those described in hemophilic arthritis in humans. Immunofluorescence studies indicated that a specific immune response is probably not involved in the pathogenesis of hemophilic arthritis. Detailed histopathologic examinations of knee joints in both the early and the late phase of arthritis revealed an obvious synovial and cartilage iron load, in the absence of inflammatory changes. The implications of these findings in the pathogenesis of destructive cartilage changes are discussed. PMID- 3048277 TI - Reactive arthritis associated with Shigella sonnei infection. AB - Several studies have failed to show an association between Shigella sonnei dysentery and reactive arthritis. We describe 3 patients who had reactive arthritis and a recent or concurrent S sonnei infection. To our knowledge, this is only the second study to suggest this association. We propose that S sonnei should be considered as a triggering agent for reactive arthritis. PMID- 3048276 TI - Suppression of streptococcal cell wall-induced arthritis by a potent protease inhibitor, bis(5-amidino-2-benzimidazolyl)methane. AB - Bis(5-amidino-2-benzimidazolyl)methane, a powerful synthetic trypsin inhibitor, proved to be highly effective in suppressing the arthritis induced by streptococcal cell wall fragments in Lewis rats. It reduced not only the degree of synovitis, osteitis, and hematopoietic hyperplasia in the distal extremities, but also the degree of associated granulomatous inflammation in the liver. The results suggest that trypsin-like proteases play an important role in this arthritis model and that inhibitors may be useful in the treatment of similar arthritic conditions in humans. PMID- 3048278 TI - Society's response to alcohol consumption and problem development. PMID- 3048279 TI - One hundred years of industrial or occupational health nursing in the United States. PMID- 3048280 TI - Dr. Harvey Knowles' contributions to the field of diabetes and pregnancy. PMID- 3048281 TI - Harvey Knowles Memorial Symposium. PMID- 3048283 TI - Pregnancy outcome in insulin-dependent diabetes: temporal relationships with metabolic control during specific pregnancy periods. AB - There are specific temporal relationships between metabolic control during different periods of pregnancy and outcome in insulin-dependent diabetic pregnancies. The rate of spontaneous abortions correlates with poor periconceptional glycemic control. Major malformations and decreased infant bone mineral content at birth correlate with poor first trimester glycemic control. Increased rates of preeclampsia and premature labor correlate with poor glycemic control in the second trimester. The development of macrosomia and that of neonatal hypoglycemia correlate with poor third trimester glycemic control. Perinatal asphyxia and neonatal hypoglycemia correlate with hyperglycemia in labor. We suggest that each stage of the pregnancy, if complicated by poor glycemic control, may lead to specific, temporally related complications. Glycemic control from the periconceptional period to the time of delivery appears important in order to minimize complications of the insulin-dependent diabetic pregnancy. PMID- 3048282 TI - Review of fetal cardiovascular and metabolic responses to diabetic insults in the pregnant ewe. AB - Studies with pregnant sheep are reviewed, which were designed to investigate whether short-term episodes of maternal hyperglycemia or hyperketonemia were detrimental to the fetus, whether ketones can cross the ovine placenta, and whether the combination of maternal hyperglycemia and hyperketonemia would contribute to an increased risk of fetal morbidity. It is concluded that acute increases in maternal ketones appear to be more detrimental to the fetus than acute increases in maternal glucose, as assessed by fetal cardiovascular, metabolic, and blood gas changes. PMID- 3048284 TI - Sources and disposition of fetal glucose: studies in the fetal lamb. AB - Three aspects of fetal glucose metabolism are studied in the pregnant sheep: source of fetal glucose, rates and routes of fetal glucose disposal, and how these processes are regulated in the fetus. However, extrapolation of findings in the fetal sheep to the condition existing in the human must be made with great caution. This is mainly due to the fact that sheep are ruminants that obtain much of their energy from volatile fatty acids and fetal sheep have low glucose and high fructose concentrations. PMID- 3048285 TI - Periconceptional metabolic status and risk for spontaneous abortion in insulin dependent diabetic pregnancies. AB - The rate of clinically apparent spontaneous abortions in insulin-dependent diabetic pregnancies has been prospectively determined to be twice as frequent as for the general population (29.5% versus 10 to 15%). In a series of several successive studies, we have shown that spontaneous abortions are associated with poor metabolic control around conception and/or in the early weeks of pregnancy, but not in the 1 to 2 weeks preceding the abortive event itself. There is also a significant relationship between decreased maternal magnesium status (as assessed by maternal serum magnesium concentration) and adverse fetal outcome (spontaneous abortion and/or major congenital malformations) in insulin-dependent diabetic women. We speculate that improvement of glycemic control and of magnesium status before conception and in the very early phases of organogenesis might improve embryonic and fetal survival. PMID- 3048286 TI - Hyaline membrane disease and surfactant protein, SAP-35, in diabetes in pregnancy. AB - Surfactant-associated protein of Mr 28,000 to 35,000 (SAP-35) is an abundant glycoprotein present in the alveolus of the lung, which imparts both structural organization to surfactant phospholipids and provides regulatory information controlling surfactant phospholipid secretion and metabolism. SAP-35 expression is enhanced by 3'-5'-cyclic adenosine monophosphate and epidermal growth factor during perinatal differentiation of type II epithelial cells. Its synthesis and RNA are also controlled by a variety of inhibitory factors, which include transforming growth factor and insulin. Glucocorticoids both enhance and inhibit SAP-35 expression in fetal lung explants. There is evidence that fetal hyperinsulinemia or hyperglycemia, or both, inhibit the morphologic differentiation of the type II epithelial cell in association with decreased phospholipid surfactant synthesis or secretion. Insulin is also a potent inhibitor of SAP-35 expression in fetal lung tissue, and decreased SAP-35 was previously noted in amniotic fluid of patients with diabetes during pregnancy. Recent progress in the management of diabetes in pregnancy, characterized by more rigorous metabolic control, has decreased the risk of hyaline membrane disease for the infant of the diabetic mother and is associated with normal levels of SAP 35 in amniotic fluid. Hyaline membrane disease remains a major cause of morbidity in infants of diabetic mothers but may also reflect a higher incidence of premature delivery, cesarean section, and asphyxia at delivery as well as inhibition of pulmonary surfactant phospholipid synthesis or expression of the surfactant protein SAP-35. PMID- 3048287 TI - Lymphocyte subpopulations during acute and convalescence phases of malaria. AB - Lymphocytes of normal healthy persons were separated from blood by Ficoll-Hypaque gradient centrifugation and iron-magnet application. peripheral blood lymphocytes (PBL) were stained by various dye-labeled monoclonal antibodies. Cells positive for specific surface markers were enumerated by a fluorescence activated cell sorter (FACS) and fluorescence microscope (FM). The results revealed that the percentages of cells positive with one monoclonal antibody counted by these two techniques were similar while the percentages of cells with double staining were higher when counted by FACS than by FM. Lymphocyte subpopulations of 18 patients infected with Plasmodium falciparum during acute and convalescence period were studied. Lymphocytopenia occurred during the acute infection while total white blood cell counts were normal. PBL of the patients were stained with OKT3, OKT4, OKT8, Leu-11 and a combination of Leu-7, Leu-1 monoclonal antibodies. The absolute numbers of all lymphocyte subpopulations were decreased during the acute infection while T8 positive cells were decreased in both percentage and absolute number. Thus T4:T8 ratio (1.7:1) became higher than normal (1.3:1) at this period. During convalescence phase, absolute numbers and percentages of Leu-7+, Leu-1+ and perhaps Leu-7+, Leu-11- cells which had low NK cell activity were significantly higher than during acute illness. The finding might explain why the NK cell activity was low during the convalescence period. PMID- 3048288 TI - The influence of microtiter plates from different suppliers on lymphocyte proliferation. AB - Microtiter plates have been popularly used for lymphocyte culture, but the influence of culture plates from different sources has not been investigated. In this study, the degree of mitogen-induced cell proliferation was investigated using six different brands of flat-bottomed plates. Lymphocytes from twelve normal donors were cultured for 96 hours with several mitogens including PHA, Con A and PWM. Spontaneous cell proliferation was slow and it did not differ significantly among the different plates. However, mitogen-induced cell proliferation showed a wide variation among the six types of plates used. The importance of selecting certain kinds of plates for specific purposes is emphasized. PMID- 3048290 TI - Medical aspects of Arctic exploration. 3. Farthest north with the "Fram". Fridtjof Nansen (1893-1896). PMID- 3048289 TI - AIDS in India: a report of three cases with review of literatures. AB - Sera from 3500 individuals including patients of non-Hodgkin's lymphoma, patients with sexually transmitted diseases, leprosy, jail inmates, healthy laboratory workers and patients on hemodialysis were screened for antibodies against human immunodeficiency virus (HIV) as a sero surveillance exercise of one of the centers set up by the India Council of Medical Research at the Postgraduate Institute of Medical Education & Research, Candigarh. Three individuals were found to be strongly positive by ELISA using the Wellcozyme kit, with no background noise in the population studies. These three patients were also evaluated by the Western blot technique and showed strong antibody reaction to all the major gene products. It was also possible to ascertain the approximate incubation period of infection in a case who rapidly developed full blown AIDS and died of wasting syndrome. Comprehensive data of 14 cases was also collected from all the centers in India and in every case one could trace the contact outside India. None of the 3 cases studied at our center had Kaposi's sarcoma. These observations could be important landmarks in the epidemiology of AIDS in India. PMID- 3048291 TI - The east Greenlanders' own traditions to social care for widows and children. PMID- 3048292 TI - Medical aspects of Arctic exploration. 4. A brave assault on the Northwest Passage: W. Edward Parry (1819-20). PMID- 3048293 TI - The early days of arthroscopic surgery in Japan. PMID- 3048295 TI - Chylothorax following high translumbar aortography: a case report and review of the literature. PMID- 3048294 TI - Arthroscopic surgery--past, present, and future. PMID- 3048296 TI - Neurological disorders associated to dengue infection. PMID- 3048297 TI - [Medical ethics and the Ethics Advisory Committee]. PMID- 3048299 TI - Human growth hormone and Creutzfeldt-Jakob disease. PMID- 3048298 TI - Colonic histoplasmosis simulating Crohn's disease in a patient with AIDS: case report and review of the literature. PMID- 3048300 TI - [Colloid osmotic pressure--physiologic, pathophysiologic and clinical significance]. PMID- 3048301 TI - [The anesthesia respirator 423141--results of an experimental and clinical trial]. PMID- 3048302 TI - [Optimizing anesthesia ventilation. 4. Anesthesia ventilation with inspiratory plateau]. PMID- 3048303 TI - [The role of inhalation chambers in the treatment of asthma]. AB - Inhalation chambers (spacers), interposed between the aerosol doser and the patient's mouth take 3 forms: lengthened tubes, large volume plastic sacs and conical chamber in the shape of a pear. Their chief advantages are: A greater proportion of the drug reaches the airways, as the deposition in the oropharynx is reduced. Co-ordination of lung and hand, a major obstacle to the correct use of aerosol-dosers, is no longer necessary. PMID- 3048304 TI - [Nasal rhinosporidiosis. Description of a case and review of the national literature]. PMID- 3048305 TI - [Pharmacokinetics of ampicillin in kidney donors and in children with kidney transplants]. PMID- 3048306 TI - The development of visual lateralization in the domestic chick. AB - In the domestic chick the visual systems fed by right and left eye differ already on day 2 in the way in which they analyse stimuli. The right eye system (RES) tends to use conspicuous cues to assign stimuli to categories whilst the LES is interested in all properties of stimuli including position in space. As early as testing is possible, the LES shows advantage in the use of topographical cues, and the RES in distinguishing food from other targets. Sharp changes in the ability of RES or LES to take charge of behaviour follow: on day 8 RES controls response even in tasks where the LES usually has advantage. Rapid change then follows: by day 10 the LES is able to control behaviour under appropriate conditions. In natural broods the sudden appearance of exploration away from the mother on day 10 may result from this change. Such periods of control by one eye system may allow intensive appropriate learning. The timing of the transition is the same in both sexes, even though females behave in almost all tasks as though the LES were less specialized (or more under the control of RES). PMID- 3048307 TI - Hemispheric differences in motor control. AB - Two lines of evidence are presented to suggest that the left hemisphere in human beings plays a special role in the organization of complex motor behaviour, an idea first put forward by Liepmann and extended more recently by Kimura. The results of one line of research suggest that the right-sided asymmetries observed in movements of the mouth during verbal and non-verbal tasks reflect the fact that mechanisms within the left hemisphere are particularly involved in selecting individual movements and facilitating the transition from one movement to another. The results of the second line of research extend this idea and suggest that the organization of eye and limb movements during visually guided reaching is also dependent on these left-hemisphere mechanisms. These findings, together with the work of a number of other workers, all point to the same conclusion: that speech is but one example of a great number of different motor patterns mediated in part by neural systems within the so-called 'dominant' hemisphere. PMID- 3048308 TI - The precursor of sulfatide activator protein is processed to three different proteins. AB - The enzymic degradation of a number of sphingolipids in the lysosomes is stimulated by small acid glycoproteins named activator proteins. We purified and sequenced a new protein, called component C, which seems to be related to sulfatide activator and to a recently described activator of glucosylceramidase (A1 activator) (Kleinschmidt, T., Christomanou, H. & Braunitzer, G. (1987) Biol. Chem. Hoppe-Seyler 368, 1571-1578). It consists of 78 amino acids and carries one carbohydrate chain at aparagine 20. Component C shows 21.5% sequence homology to sulfatide activator and 34.2% homology to A1 activator. Structural similarities between these three proteins have also been detected. Recently the cDNA sequence of the sulfatide activator precursor has been published (Dewji, N.N., Wenger, D.A. & O'Brien, J.S. (1987) Proc. Natl. Acad. Sci. U.S.A. 84, 8652-8656). We could align the protein sequences of sulfatide activator, A1 activator and component C with that of this large precursor protein. After minor corrections of the DNA sequence we obtained total fit. Thus it seems that three different proteins are derived from the sulfatide activator precursor by proteolytic processing. Possible processing sites were found on the precursor at sites adjacent to the N-termini and C-termini of the mature proteins. The processing of sulfatide activator was studied by Fujibayashi and Wenger (Fujibayashi, S. & Wenger, D.A. (1986) Biochim. Biophys. Acta 875, 554-562). Their data support our assumption that processing occurs by simultaneous cleavage at all possible sites. PMID- 3048309 TI - Endogenous and exogenous TNF therapy (EET therapy): conceptual and experimental grounds. PMID- 3048310 TI - The anti-cancer activity of tumour necrosis factor. PMID- 3048311 TI - Necrotizing activity of tumour necrosis factor and its mechanism. PMID- 3048313 TI - Effects of TNF in bacterial infections. PMID- 3048314 TI - Mechanisms and significance of the mitogenic and antiviral actions of TNF. PMID- 3048315 TI - The pathophysiologic role of cachectin/TNF in septic shock and cachexia. PMID- 3048312 TI - Multiple facets of induction of tumour necrosis. PMID- 3048316 TI - TNF as an activator of vascular endothelium. PMID- 3048317 TI - Sensitizing effects and mechanisms of desensitization in regulation of the in vivo response to TNF. PMID- 3048318 TI - Protection against fatal Klebsiella pneumoniae sepsis in the squirrel monkey Saimiri sciureus after immunization with a capsular polysaccharide vaccine. AB - An anti-Klebsiella pneumoniae K5a capsular polysaccharide vaccine was evaluated as a preventive approach for protecting squirrel monkeys, Saimiri sciureus, in our breeding colony. Based on an 8-month vaccination schedule over a period of more than two years, this vaccine, regardless of the animal's age, resulted in a reduction of this particular bacterial infection and its generally associated fatal outcome. IgG antibody responses in naive and vaccinated animals were monitored over an extended period by radioimmunoassay and showed a marked increase above the initial naturally occurring antibody titres. No side-effects were observed after repeated vaccination of several hundred animals during a two year period. PMID- 3048319 TI - Aspects of haemopoietic cell dynamics: ontogeny and targeted migration. AB - In the developing avian and mammalian embryo, haemopoietic cells appear first in transient foci whose function is restricted to discrete periods of embryogenesis. These foci are essentially represented by the yolk sac, intraembryonic dispersed foci and the liver. Haemopoietic cells then repopulate the developing spleen, thymus and bone marrow, organs which persist and develop after birth. In the present review, we describe a number of possible mechanisms controlling specific adhesion, oriented migration and invasiveness of haemopoietic cells. One concerns the high specificity of the interactions of homing receptors on the surface of haemopoietic cells with determinants on vascular endothelium and/or thymic epithelium. A second is the importance of the presence of some macromolecules in the extracellular matrix, such as fibronectin, collagen, laminin and elastin. These components can interact with the haemopoietic cells (and/or induce chemotaxis) via the existence of specific receptors on the surface of the haemopoietic cells. Another mechanism is the activation of the haemopoietic cells through the interactions of cell-chemotactic factor, cell-extracellular matrix and/or cell-thymic epithelium. This activation can lead to: 1) the expression of new specific cell-surface receptors for the target foci; 2) the secretion of specific protease and glycosidase systems active upon the extracellular matrix; and 3) the differentiation of these cells in the thymus. PMID- 3048320 TI - Activated lymphocyte cytotoxicity: concepts and confusions. PMID- 3048322 TI - Characterization and functions of natural killer cells. PMID- 3048321 TI - NK cells: a discrete leukocyte subset of still undefined origin mediating biologically relevant functions upon specific stimulation. PMID- 3048323 TI - Can acquired immunodeficiency syndrome and Creutzfeldt-Jakob disease be transmitted via otologic homografts? AB - Materials commonly employed in the preparation of otologic homografts such as ethanol and formaldehyde are effective in vitro in inactivating human immunodeficiency virus (HIV). However, to our knowledge, the complete permeation of homograft materials with preservative has not been demonstrated. Ethanol and formaldehyde have not been shown to be effective in inactivating the Creutzfeldt Jakob agent. The literature on sterilization procedures for these agents is reviewed. Standard procedures for preparation of otologic homografts are examined. It is recommended that donor HIV serologic status be determined when otologic homografts must be used. Further research is required to determine the efficacy of otologic homograft sterilization techniques against HIV and Creutzfeldt-Jakob disease. PMID- 3048324 TI - Beta 2-microglobulin in otitis media with effusion. AB - beta 2-Microglobulin (beta 2M) is a low-molecular-weight protein present in serum and other fluids during various autoimmune or chronic inflammatory diseases. beta 2-Microglobulin was measured in middle ear effusion (MEE) and serum samples obtained from 36 patients with chronic otitis media with effusion. Using a quantitative competitive enzyme immunoassay, we were able to demonstrate beta 2M in 98% of the MEE samples. The mean concentration of beta 2M was higher in the MEE samples than in the serum samples. There was considerable variability between ears in those patients with bilateral MEE. There was no correlation between beta 2M concentration and the patients' age, sex, MEE type, and culture results, or cytologic profiles of the MEEs. This increased beta 2M may reflect earlier lymphocyte activity during the inflammatory process. PMID- 3048325 TI - Control models of cell cycle transit, exit, and arrest. AB - Several distinct cycles mediate the events which occur between one cell division and the next. In micro-organisms there are generally two cycles. One governs biomass growth, the other DNA synthesis and cell division. In higher eukaryotes there can be as many as four distinct cycles, with growth, DNA synthesis, cell division, and nuclear division each possessing its own functional sequence of events. These cycles are controlled and coordinated by several different regulatory mechanisms. Restriction points are specific steps in the cycle whose completion is governed by external regulatory agents. One set of restriction points requires nutrients and growth hormones for step completion. Another set serves as receptors for differentiating factors which cause cycle arrest and initiate cellular differentiation. There is currently a debate as to whether restriction point inhibition involves permanent arrest or temporary arrest with a stochastic arrested-state residence time controlled by a transition probability mechanism. Tissue sizing is a process of negative feedback inhibition mediated by intercellular communication via cell surface contact and the extracellular matrix. Sizers commonly operate throughout broad portions of the cycle and appear to cause a slowing of cycle transit velocity rather than arrest. Sizers are probably the major regulatory mechanism for cell growth under conditions of nutrient and growth factor excess. They also generate compensatory proliferation following wounding or cell death. A growing body of evidence suggests that both the transit velocity, with which cells move through their several cycles, and the coordination of the cycles are controlled by intracellular regulatory mechanisms which behave as biological oscillators. These oscillators trigger complex sequences of events such as DNA synthesis and cell division. PMID- 3048328 TI - The roles of epithelial-mesenchymal cell interactions in developmental processes. AB - This review surveys some recent trends in the study of the developmental interactions between epithelial and mesenchymal cells. The influence of such interactions on cell differentiation is considered with reference to kidney development, limb bud development, chondrogenesis and osteogenesis, and tooth development. Effects on epithelial morphogenesis are discussed, using salivary gland development as an example. The roles of humoral factors (hormones or growth factors) are considered, and the evidence for the participation of cell adhesion molecules is examined. PMID- 3048326 TI - Differentiation, cancer, and anticancer activity. AB - Carcinogenesis is a multistep process that results from the development of a variety of defects in the control of differentiation and proliferation. To investigate this concept further, 3T3 T mesenchymal stems cells were employed to establish that a distinct sequence of biological processes is involved in the control of differentiation and proliferation, and that these processes are integrally regulated. Specific defects in these regulatory processes were next established as being involved in carcinogenesis. These defects, however, were found not to be absolute; rather, they appear to involve changes in the stringency by which differentiation and proliferation are integrally regulated. Finally, it was established that when normal or transformed stem cells are induced to undergo nonterminal differentiation (which is one step in the integrated control of proliferation and differentiation), they can be made resistant to carcinogenesis or to revert to a nontransformed state. These data provide strong evidence that a critically important requirement for normal homeostasis is maintenance of intact cellular mechanisms to integrally regulate differentiation and proliferation. PMID- 3048327 TI - The regulation of mitotic spindle function. AB - The process of mitosis includes a series of morphological changes in the cell in which the directional movements of chromosomes are the most prominent. The presence of a microtubular array, known as the spindle or mitotic apparatus, provides at least a scaffold upon which these movements take place. The precise mechanism for chromosome movement remains obscure, but new findings suggest that the kinetochore may play a key role in chromosome movement toward the spindle pole, and that sliding interactions between or among adjacent microtubules may provide the mechanochemical basis for spindle elongation. The physiological regulation of the anaphase motors and of spindle operation either before or after anaphase remains equally elusive. Elicitors that may serve as controlling elements in spindle function include shifts in cytosolic calcium activity and perhaps the activation or inactivation of protein kinases, which in turn produce changes in the state of phosphorylation of specific spindle components. PMID- 3048329 TI - Host binding proteins and bacterial adhesion: ecology and binding model. AB - Defining the involvement of specific recognition and (or) adhesion molecules in the precise association formed between cells of an organism during development or between bacteria and specific host tissues has become a focus of extensive research. The possibility that the same molecules responsible for cellular adhesion in the host may also play a major role in determining host-bacterial interactions is now becoming more evident. The following review looks at the interaction of a group of host binding proteins, including lectins, fibronectin, and laminin, with respect to their specific association with bacteria. This information is dealt with both from the perspective of the ecology of the host and its autochthonous and pathogenic bacterial populations, as well as in terms of the difficulties in defining the nature of ligand associations even in the more simplified bacterial-host interaction. PMID- 3048330 TI - Integration of signal-transduction processes. AB - The adenylate cyclase - cAMP, phospholipase C - IP3 (inositol 1,4,5 triphosphate), and DAG (diacylglycerol) signal transduction systems are used to illustrate general principles underlying the process of information transfer during cell stimulation. Both systems consist of reaction cascades that convert the external signal to an intracellular messenger, translate the messenger to regulatory activities, and then modulate the activities of appropriate cellular proteins to result in specific cell responses. Almost all of these reactions are under second-messenger-dependent regulation, with many being regulated by multiple messengers. Such complex regulation provides ample opportunities for the fine-tuning of the signal cascades and for coordination between cascades during cell stimulation. Specific examples are used to illustrate how the cell uses different intrasystem and intersystem regulatory reactions to achieve specific responses. PMID- 3048331 TI - Transactivation by the adenovirus E1A gene. AB - The 289aa product of the adenovirus E1A gene mediates the transcriptional activation of the set of early viral genes as well as several cellular genes. The E1A protein is not a DNA binding protein but, rather, acts indirectly to achieve the activation. The process of viral gene activation involves the use of cellular transcription factors, and in at least one case, in vivo assays have demonstrated a stimulation of stable promoter complex formation as a function of the E1A gene product. Analysis of transcription factors in nuclear extracts has identified a cellular factor, termed E2F, with specificity for the viral E2 promoter. The concentration of this factor increases as a result of the action of E1A. This increase in DNA binding activity does not require protein synthesis, thus indicating an E1A-mediated modification of a pre-existing factor. The E2F factor has been purified to homogeneity and is a polypeptide of 54,000 molecular weight. Analysis of an additional viral promoter, the E4 promoter, has identified a protein that interacts with sequences critical for transcription. This factor, termed E4F, is also increased as a function of the E1A product. The E4F factor has also been purified to homogeneity and has a molecular weight of 50,000. Therefore, the coordinate control of transcription by the E1A gene product involves the activation of multiple promoter specific factors. PMID- 3048332 TI - Transcriptional activation of heat-shock genes in eukaryotes. AB - Prokaryotes and eukaryotes respond to thermal or various chemical stresses by the rapid induction of a group of genes collectively referred to as the heat shock genes. In eucaryotes, the expression of these genes is primarily regulated at the transcriptional level. The early observations that transfected heat shock genes were inducible in heterologous systems suggested the existence of common regulatory elements in these ubiquitous genes. Sequence analysis of cloned Drosophila heat shock genes revealed a conserved 14 base pair (bp) inverted repeat, which is essential for heat induction. This regulatory sequence, referred to as the heat shock element (HSE), is found in multiple imperfect copies upstream of the TATA box of all heat shock genes. While studies in heterologous systems indicated that a single copy of HSE was sufficient for inducibility, further analysis in homologous assays suggests that multiple HSE can act in a cooperative way and that the efficiency of transcriptional activation is related, within limits, to the number of HSE. Comparative analysis of heat shock genes reveals that HSE can be positioned at different distances from the TATA box in either orientation, a behavior reminiscent of enhancer elements. However, the presence of HSE does not necessarily confer heat inducibility, as shown by their presence in the constitutively expressed but non-heat-inducible homologous cognate genes. Footprinting and nuclease mapping have been used to show that a protein factor (HSTF: heat shock transcription factor) binds to the HSE element, activating heat shock gene transcription in a dose-dependent manner. The recent progress in the isolation and characterization of HSTF in Drosophila, yeast, and human cells is reviewed. Finally, different models suggested to account for the positive regulation of heat shock genes by the HSTF are presented. PMID- 3048333 TI - DNA-mediated transfection as a model for oncogenesis. AB - DNA transfer technology has greatly contributed to progress in understanding molecular biology and genetics. In recent years, great efforts have been expended to determine the oncogenic potential of single, defined genes or complex gene mixtures as a prelude to defining the role those genes may play in neoplastic transformation in vitro and tumor induction in vivo. This paper reviews the currently available DNA transfection techniques and their application toward understanding cancer initiation and progression, and how the in vitro and animal models may apply to human cancer. PMID- 3048334 TI - Interaction of tumor cells and immune system in the metastatic process. AB - The metastatic process is rather complicated and relatively inefficient. Millions of tumor cells are constantly shedding from the primary tumor into the blood stream, but very few of them are able to form metastatic tumors in the different organs or tissues of the host. It is widely accepted that metastatic cells have to possess a complex array of various properties that allow them to complete the metastatic cascade. The realization of the metastatic potential largely depends on the ability of tumor cells to evade host defense mechanisms. The potential role of specific and nonspecific immune mechanisms in the control of metastatic spread and growth is the subject of the present review. A better understanding of the mechanisms of antimetastatic defense is of prime importance for development of efficient immunotherapeutic methods for the treatment and eradication of disseminated tumor metastases. PMID- 3048335 TI - The production of tissue-specific histone complements during development. AB - At least two mechanisms generate tissue differences in the histone subtype composition during development: subtype dilution and subtype replacement. Subtype dilution, which occurs when cells continue dividing after having ceased to synthesize one more histone subtypes, allows the elimination of stable subtypes. It is the major mechanism generating cell differences in histone composition in sea urchin embryogenesis. Subtype replacement has been observed in mammalian tissues, both in the intact animal and in cultured cells. It is most evident in nondividing cells but occurs to some extent in dividing cells as well. Examples of the two mechanisms are presented and their possible biological significance, as well as that of the differences they produce, is discussed. PMID- 3048336 TI - Somatosensory evoked potentials by median nerve stimulation in rabbits. I. Source localization by cortical and stereotaxic recordings. PMID- 3048337 TI - [Rapid diagnosis of viral diarrhea]. PMID- 3048338 TI - Serum aminoterminal type III procollagen peptide. Relation to biosynthesis of collagen type III in experimentally induced granulation tissue in rats. AB - Serum aminoterminal type III procollagen peptide was measured in rats during the development of granulation tissue induced by subcutaneous implantation of viscose cellulose sponges. Active collagen type III synthesis in granulation tissue during the first three weeks was accompanied by an increase in serum propeptide level. A positive correlation was observed between the increase in serum propeptide level on the one hand and the increase in granulation tissue collagen type III content and the in vitro formation of tissue 3H-hydroxyproline on the other hand. In some animals the serum propeptide level remained low, despite biochemical signs of collagen synthesis, indicating variations in the release into serum and/or the metabolism of circulating propeptide. The increase in propeptide antigen concentration was mainly due to an elevated content of material with molecular weight equal to or twice that of the propeptide. A minor fraction of the propeptide remained attached to the interstitial collagen fibres in the granulation tissue. The correlation between the serum propeptide level and the biosynthesis of collagen at the site of the focal fibroproliferative process suggests that the serum propeptide level may be a valuable indicator of fibrogenesis and thereby of disease activity in fibrotic conditions. PMID- 3048339 TI - Premalignant and malignant oral lesions are associated with changes in the glycosylation pattern of carbohydrates related to ABH blood group antigens. AB - The distribution of carbohydrate structures related to the ABO(H) blood group antigen system was studied in biopsies from eight squamous cell carcinomas, and eight erythroplakias with epithelial dysplasia. Twenty oral lesions without histological evidence of malignancy (13 lichen planus lesions and 7 homogeneous leukoplakias) were also examined. The distribution of Lex, Ley, H type 2 chain, and N-acetyllactosamine, all type 2 chain carbohydrate structures, was investigated by immunohistological staining using monoclonal antibodies with selected specificity. The histological pattern of expression of these antigens in the benign lesions was similar to that of normal oral mucosa, i.e. expression of: N-acetyllactosamine on basal cells, H antigen on parabasal cells, and Lex and Ley on spinous cells. However, lesions with epithelial dysplasia showed H antigen on all spinous cells, and often also on basal cells, with expression of Lex and Ley restricted to the most superficial part of the epithelium above the H-positive cell layers. In carcinomas most cells were negative for H antigen but were positive for Ley and Lex in 5 out of 8 cases. PMID- 3048341 TI - Silver jubilee of the British Endodontic Society. PMID- 3048342 TI - A clinical survey of failed post retained crowns. PMID- 3048340 TI - Age-related changes in vertebral trabecular bone architecture--assessed by a new method. AB - Cylindrical trabecular bone specimens (d = 7 mm) were drilled in a vertical direction from the central part of the third vertebral body (L3) from 23 normal individuals aged 15-87 years (10 males and 13 females). The bone samples were embedded in methylmetacrylate and sawn in 400 microns thick sections with an arbitrary rotation but a fixed vertical axis. The sections were investigated in polarized light at a magnification of x 8. By using this technique, vertical and horizontal trabeculae were clearly separated due to different colors. Photographs were taken. These were magnified, and trabecular thickness and intertrabecular distance were measured, using a Zeiss-integration plate II. A significant age related decrease was found in the mean horizontal trabecular thickness (r = 0.71, p less than 0.001), while the mean thickness of the vertical trabeculae was unchanged with age (r = 0.06, n.s.). Furthermore, a significant increase was found both for the mean distance between the horizontal trabeculae (r = 0.79, p less than 0.001) and between the vertical trabeculae (r = 0.75, p less than 0.001). The present study gave a clear and striking visual presentation of both the thinning and disappearance of the horizontal supporting struts in the vertebral trabecular lattice and the total removal of some of the vertical trabeculae--leading to the dramatic loss of bone strength previously demonstrated. PMID- 3048343 TI - Temperature changes of materials during impression taking. PMID- 3048344 TI - Vomiting during the taking of dental impressions. Two case reports of the use of psychological techniques. PMID- 3048345 TI - Optical spectroscopy. Pre-mammography marker. AB - Non-invasive optical spectroscopy consistently delineates compositional and physiologic properties of breast tissues serving as a pre-mammography risk marker for cancer or yielding a high assurance of no such risk. We believe this new non imaging approach depends on biochemistry of tissues rather than on the macroscopic physical properties involved with most breast imaging modalities. After establishing the procedure as inexpensive, physician independent, simple, requiring only a few minutes and appealing to women, it was carried out in two institutions on 1739 women referred for routine mammography. Of 166 breast biopsies on these women 77 were cancer by histology. An automated computerized analysis of the spectroscopic data yielded a sensitivity of 87 per cent, a specificity of 74 per cent and a negative predictive value of 99 per cent. Optical spectroscopy shows promise in identifying women at a higher risk for developing cancer, cases of non-infiltration carcinomas where dense breasts limit mammographic detection, and even clustered calcifications not associated with a mass. The relative risk of breast cancer was 16.5 times as great with a positive spectroscopic value at a sensitivity range of 87 per cent. Placement of 87 per cent of all breast cancer cases in a subset of 28.7 per cent of all women will yield a population of women in whom mammography will be approximately four times as efficient. PMID- 3048347 TI - Angiomyolipoma of the kidney. Comparison between magnetic resonance imaging, computed tomography, and ultrasonography for diagnosis. AB - In six patients with angiomyolipoma of the kidney, five showed a typical pattern in T1, proton and T2 weighted images, with magnetic resonance imaging (MRI). Angiomyolipoma is recognized by its characteristic fatty tissue components, which result in high signal intensity in T1 and proton weighted images and a low signal in T2 weighted images. Five of the patients were also examined with ultrasonography (US); in four of these, the tumor could be recognized by its echodense pattern. Computed tomography (CT) was slightly less sensitive; three out of six examinations failed to produce a definite diagnosis. Instead of the typical low attenuation values of fat, Hounsfield units similar to either muscle or kidney tissue were observed in these 3 cases. In conclusion, the specific diagnosis of angiomyolipoma depends on the amount of fatty tissue present at MRI, CT, and sonography. PMID- 3048346 TI - Nine pheochromocytomas in the same patient. Final mapping with ultrasound and angiography. AB - A 37-year-old man presented with hypertension and elevated urine catecholamine. Ultrasound scanning revealed a solid tumour of the right adrenal gland and two solid tumours in the retroperitoneum. The findings were confirmed with computed tomography and abdominal angiography. At surgery only the tumour of the right adrenal gland was removed. The histopathologic diagnosis was pheochromocytoma. Postoperatively the symptoms and biochemistry were unchanged and the patient was referred for further treatment. At ultrasonography and abdominal aortography 6 remaining tumours were demonstrated. Surgery was performed and 8 pheochromocytomas were extirpated (3 were closely spaced small tumours in a conglomerate corresponding to one of the visualized tumour sites). On histopathologic examination no signs of invasive growth were found. The patient recovered completely. The blood pressure was still normal 2 1/2 years later. Angiography and non-invasive examination of the entire abdomen and pelvis should be routine when pheochromocytomas are searched for. PMID- 3048348 TI - Renal medullary cystic disease. Findings at urography and ultrasonography. AB - Medullary cystic disease (MCD) is an uncommon renal disease with adult onset and autosomal inheritance, eventually progressing to terminal renal failure. It may be difficult to identify because of insufficient diagnostic tools. At urography, the same ring-shaped accumulation of contrast medium at the corticomedullary junction was observed in two patients (mother and son) suffering from MCD. To our knowledge this observation has not been reported before. PMID- 3048349 TI - Reliability of ultrasound-guided fine-needle biopsy of pancreatic masses. AB - Fine-needle aspiration biopsy (FNB) was performed with ultrasound guidance in 79 patients in whom sonography had revealed a mass suggesting pancreatic malignancy. The final diagnosis (surgery, autopsy and clinical course) in 69 of these 79 patients was a malignancy closely related to the pancreas while in the remaining 10 patients benign disease was confirmed. A correct diagnosis of malignancy was attained by FNB in 59 of the 69 patients with a malignant tumour while in 10 it failed to confirm the diagnosis. FNB yielded a true negative result in 10 patients with benign disease. The accuracy of sonographically guided FNB in the present investigation was 87 per cent. Ultrasound-guided fine-needle biopsy is considered the method of choice for further evaluation of pancreatic masses. PMID- 3048350 TI - Hypoechoic lesions without halo in echogenic liver. A frequent sonographic dilemma. AB - Sonographic evaluation for the presence of hypoechoic hepatic lesions without halo was carried out in 365 consecutive patients with echogenic livers. In 115 patients (31%) such lesions could be demonstrated. Computed tomography of the liver was performed in 52 of these patients, a long term sonographic follow-up in 76, and a biopsy in 3 cases. In 103 patients the hypoechoic lesions were due to sonographic pseudolesions (PL's), probably representing normal liver tissue in otherwise diffusely fatty infiltrated livers. The PL's showed characteristic sonographic appearances such as a missing mass effect, a 'landscape'-like configuration with angulated margins and slender extensions of hypoechoic tissue. The PL's were located below the capsule, near the gallbladder (41%), and ventral to the portal vein (37%). In 75 per cent they occurred in a liver with considerably increased echogenicity. In 12 patients hypoechoic lesions were caused by circumscribed malignant or infectious involvement of the liver. They could be discriminated from PL's by their mass-like appearance in 8 subjects. In 4 of these 12 cases the foci were of PL-typical appearance, but not of PL-typical location. In the light of these results and of recently published reports a rational diagnostic approach to hypoechoic lesions without halo in echogenic livers varies, depending on such factors as known primary malignancy or site of the lesion. PMID- 3048351 TI - Sterno-costo-clavicular hyperostosis. A case report with a review of the literature. AB - The clinical and radiologic findings in a case of sterno-costo-clavicular hyperostosis are reported and compared with the findings in the 23 Caucasian and about 300 Japanese cases reported in the literature. The main complaints are pain in the upper anterior chest wall and sometimes limited mobility of the shoulders. Radiologically, the clavicles, the sternum and the first ribs are grossly enlarged with complete fusion between them. As reported in previous cases, our patient had conspicuous congestion of the external jugular veins, but no other signs of compression in the thoracic inlet. There was asymptomatic compression of both subclavian veins, but none of the previously reported skin manifestations and no complaints from other parts of the locomotive system. The patient was HLA B27 negative. PMID- 3048352 TI - Structure of the met protein and variation of met protein kinase activity among human tumour cell lines. AB - An in vitro autophosphorylation assay has been used to demonstrate that there is considerable variation in met associated protein kinase among human tumour cell lines. Of particular note was the very high level of autophosphorylation of the 140 kD met protein (p140met) in experiments with A431 human cervical carcinoma cells. In contrast in experiments with Daoy human medulloblastoma cells we failed to detect phosphorylation of p140met; instead a high level of phosphorylation of a 132 kD protein was observed. To help understand the basis for the variation in kinase activity and to learn more about the structure of the mature met protein we have analysed p140met in SDS-polyacrylamide gels under non-reducing conditions. Under these conditions the met protein had an apparent molecular weight of 165,000 indicating that the mature met protein may exist as an alpha beta complex in which p140met (designated the beta subunit) is joined by disulphide bonds to a smaller, 25 kD, alpha-chain. We have identified a potential proteolytic cleavage site with the sequence Lys-Arg-Lys-Lys-Arg-Ser at amino acids 303-308 in the human met protein that may account for cleavage of the met protein into alpha and beta subunits. PMID- 3048353 TI - A phase 1 and pharmacokinetic study of didox: a ribonucleotide reductase inhibitor. AB - A phase 1 study of a new ribonucleotide reductase inhibitor, didox, was performed by administration of escalating doses of the drug by slow i.v. injection. Thirty four patients with unresponsive metastatic carcinoma received the drug. There were 13 escalations of dosage, from a starting dose of 192 mg m-2 to 10 g m-2. Dose limiting toxicity was encountered at 7.5 g m-2 where disturbances of hepatic and renal function were observed, in addition to severe gastrointestinal toxicity. Pharmacokinetic studies showed that a peak level of didox was achieved within 5 minutes of injection. At 1,728 mg m-2 the data best fitted a 2 compartment open model, with a mean serum alpha t1/2 of 5.2 min, with a beta t1/2 of 41.3 min. Less than 10% of the drug was excreted unchanged in the urine and the majority of this excretion was within 6 h. Didox can therefore be safely given by slow i.v. injection at a dose of 6 g m-2. PMID- 3048354 TI - Cultivation of human breast carcinoma in soft agar. Experience with 237 fresh tumour specimens. AB - A total of 237 breast carcinomas have been studied with the Courtenay-Mills (C-M) soft agar method. Cell yields and plating efficiencies (PE) were recorded after various enzyme treatments. The highest cell yields and PEs were obtained with the combination of collagenase 0.5%, hyaluronidase 1000 IE ml-1 and DNase 0.1% and an incubation time of 2 h. Eighty percent of the specimens gave greater than 10 colonies, and 60% formed greater than 30 colonies permitting chemosensitivity studies. The C-M method gave significantly higher PEs than the Hamburger-Salmon (H-S) method. Hormone supplements (insulin, oestradiol, progesterone, hydrocortisone) and also reduced agar concentrations (less than 0.3%) gave marginal stimulation of colony formation. In chemosensitivity studies involving doxorubicin, vincristine and 4-OOH-cyclophosphamide, the C-M method gave dose response relationships without plateaus. PMID- 3048356 TI - Rhinitis and sinusitis. PMID- 3048357 TI - Getting a patient off the ventilator. PMID- 3048358 TI - Selman Waksman (1888-1973), discoverer of streptomycin: a centenary review. PMID- 3048359 TI - Chronic obstructive lung disease prevention. PMID- 3048355 TI - Recessive mechanisms of malignancy. AB - It is increasingly recognised that recessive mutations play an important role in the pathogenesis of many forms of malignancy. Some of the affected loci may prove to be recessively-activated proto-oncogenes, but others are now known to be tumorigenic solely by virtue of their loss or inactivation and therefore form a distinct and novel family of tumour genes. Preliminary evidence suggests that such genes are likely to be functionally heterogeneous and to encode molecules involved in the inhibition of cellular proliferation and/or the induction of differentiation. Their further study is likely to illuminate fundamental mechanisms of normal cellular growth and differentiation as well as having important implications for the pathogenesis and management of cancer. PMID- 3048360 TI - Pulmonary involvement in ulcerative colitis. AB - Pulmonary involvement in ulcerative colitis usually presents as a rapidly progressive cough with copious amounts of sputum. Although it is rare, distressing symptoms may be relieved by inhaled steroids in 50-60% of cases. Three case reports are presented along with a review of the features of 28 other cases. PMID- 3048362 TI - Chest physiotherapy: time for reappraisal. AB - Chest physiotherapy should now be updated with attention to three important features: first, its use should be limited to those patients with actual or potential sputum production and its central aim should be to increase expectoration. Second, it should incorporate the forced expiration technique with postural drainage and omit traditional elements such as percussion and vibration. Third, the additional use of inhaled adrenergic agents and possibly oral high frequency oscillation may increase sputum clearance further. PMID- 3048361 TI - Stopping patients smoking. PMID- 3048363 TI - Evaluation of directed coughing in cystic fibrosis. AB - Supervised directed coughing was compared to conventional physiotherapy (postural drainage, vibration and/or percussion and coughing) in 38 patients with cystic fibrosis aged 9-18 years admitted to hospital with an exacerbation of their pulmonary symptoms. Assessment included objective measures of pulmonary function and sputum characteristics. Both treatment groups showed significant improvement at the end of the 2-week period. When the patients were graded according to their pulmonary disease, those with mild-moderate disease demonstrated a significant improvement in both treatment groups whereas those with severe lung disease showed little improvement with either treatment. Directed coughing is as effective as conventional physiotherapy in the management of patients with cystic fibrosis admitted to hospital for treatment of an exacerbation of their pulmonary symptoms. PMID- 3048365 TI - The use of high dose inhaled beclomethasone dipropionate as a means of assessing steroid responsiveness in obstructive airways disease. AB - The use of high dose inhaled beclomethasone dipropionate (BDP) as a means of assessing steroid responsiveness in chronic obstructive airways disease (COAD) was assessed in 22 patients in a study involving three consecutive 2-week periods of placebo, inhaled BDP (1500 micrograms daily), and oral prednisolone (30 mg daily). Five of the 22 patients were considered steroid responsive following the course of oral corticosteroids and in each case this responsiveness had been identified by inhaled BDP therapy. It is concluded that high dose inhaled BDP (1500 micrograms daily) may be used to assess corticosteroid responsiveness in COAD. PMID- 3048367 TI - Immotile-cilia syndrome with azoospermia: a case report and review of the literature. AB - A case of immotile-cilia syndrome associated with azoospermia is presented. This diagnosis is based on a typical history of bronchitis, sinusitis, situs inversus, impaired nasal mucociliary clearance and characteristic ultrastructural defect in the respiratory tract cilia and in the sperm tail. Semen analysis showed azoospermia with no evidence of obstruction in the epididymis or vas deferens; there was normal spermatogenesis. PMID- 3048366 TI - Aneurysmal bone cyst of the rib: a review and report of two cases. AB - Aneurysmal bone cyst (ABC) is a benign lesion and generally occurs in the long bones and vertebral column. ABC of the rib is an uncommon entity. Two cases of ABC involving the rib are reported. Its occurrence in the eight decade of life as manifested in one of our patients is extremely rare. The aetiology, clinical manifestations, pathology and treatment are briefly discussed. En bloc resection of the lesion is curative, and offers a good cosmetic and functional result. PMID- 3048364 TI - A short exercise and living course for asthmatics. AB - A 5-day, non-residential exercise and living course for children with asthma is described as a feature of a programme of outpatient physiotherapy. Eleven children undertaking such a course were compared with 10 asthmatic children in a control group. The subject group showed, in the short term at least, an improvement in bronchial lability, peak flow rates, nocturnal and daytime wheeze, and activity compared with the controls. These findings were statistically significant. There was no difference between the groups in the number of days on which extra medication was taken. A short, sharp course is of benefit physically, socially and psychologically to children with asthma. PMID- 3048368 TI - A clinicopathological study of mucosal involvement in linear IgA disease. AB - Mucosal involvement in linear IgA disease was assessed clinically and immunologically using direct and indirect immunofluorescence (IF) techniques. There was clinical evidence of oral mucosal involvement in all 10 patients examined and conjunctival diseases in six. Direct IF findings correlated well with clinical oral disease, with all patients demonstrating linear IgA deposits in the basement membrane zone of oral mucosa. However, this was not true of conjunctiva where no linear IgA could be demonstrated. Conjunctiva did provide a good substrate for indirect immunofluorescence using patients' sera, and showed that five of the 10 patients had circulating anti-basement membrane zone IgA. PMID- 3048369 TI - Improvement of psoriasis by a topical vitamin D3 analogue (MC 903) in a double blind study. PMID- 3048370 TI - Plasma levels of beta 2-microglobulin in psoriasis. PMID- 3048371 TI - Failure of isotretinoin to control dermatitis herpetiformis and subcorneal pustular dermatosis. PMID- 3048372 TI - Maternal blood viscosity and uteroplacental blood flow velocity waveforms in normal and complicated pregnancies. AB - According to the Poiseuille-Hagen law, viscosity influences flow resistance. A possible effect of blood viscosity upon the resistance index of the uteroplacental circulation as measured by continuous wave Doppler ultrasound was investigated in 50 pregnant women. It was found that blood viscosity variables explained only about 10% of the variation in the resistance index in all patients, which was not statistically significant. It is suggested, therefore, that the vascular contribution to flow resistance may be more important. PMID- 3048373 TI - Energy supplementation in the last trimester of pregnancy in East Java: I. Effect on birthweight. AB - The effect of two levels of energy supplementation in the last trimester of pregnancy on birthweight was tested in a controlled randomized trial in three villages in Madura, East Java. The high and low energy supplements provided 1.95 MJ (465 kcal) and 218 kJ (52 kcal) per day respectively. In the baseline period the home diet provided on average 6.28 MJ (1500 kcal) (SD 2.1 MJ (499 kcal] and 41 g (SD 13 g) of protein. The mean birthweight was 2835 g and the rate of low birthweight 12.2%. In the experimental period the home diet was better. The average intake ranged from 6.45 to 7.19 MJ (1541-1717 kcal) and 41.4-44.2 g per day, depending on the degree of compliance. Mean birthweight increased by 100 g and the rate of low birthweight dropped to 9.5%. There was no difference between the high and low energy supplemented group as a whole, probably due to the masking effect of the better home diet in the experimental period. It is likely that a positive effect of energy supplementation on birthweight was restricted to the group of pregnant women with the lowest home dietary intake and/or a low prepregnant weight. In this community targeting of supplementation to lean seasons and/or to women with a low prepregnant weight may be cost-effective. PMID- 3048374 TI - Ovarian size in postmenopausal women. AB - Ovarian volumes have been determined by pelvic ultrasonography in 2246 apparently healthy postmenopausal women of whom 2221 were included in the statistical analysis. Factors associated with gonadal size have been identified, and reference ranges for derived indices have been determined for use (in association with criteria for abnormal morphology) in a screening programme for ovarian carcinoma. The right ovary was present in 98.9% of subjects and the left in 99.1%. The mean (SD; range) of right and left ovarian volumes were 3.58 (1.40; 1.00-14.01) and 3.57 (1.37; 0.88-10.9) ml respectively. Significant predictors of ovarian volume were years since the menopause, weight, parity, age at menopause, a history of hormone replacement therapy, and previously diagnosed breast cancer. Abnormal ovarian volumes were assessed from a score equal to the (observed mean log volume (MLV) minus the predicted MLV)/0.327. A simplified nomogram has been prepared for routine clinical use. The relative abnormality of one ovary was assessed from a ratio score equal to loge (larger ovarian volume/smaller ovarian volume)/0.211 compared with the 99th centile for the Gaussian distribution. PMID- 3048375 TI - Inflammatory cell infiltration and survival in squamous cell carcinoma of the vulva. AB - The prognostic significance of tumour infiltration by inflammatory cells in squamous cell carcinoma of the vulva was examined in a cohort of 34 patients surviving without recurrence for at least 5 years after radical surgery, and a comparative cohort of 35 patients who died of their diseases. Overall, heavy inflammatory infiltration correlated with a good prognosis and light infiltration with a poor one, independent of other indices such as differentiation, tumour size and nodal status. IgA-containing cell infiltration also correlated with a good prognosis but the presence of IgA-containing cells did not alone account for all the inflammation in the good prognosis group. An immunological response to the tumour may be influencing prognosis. At a practical level, the extent of inflammation appears, at least in this material, to be as useful a prognostic index as many more conventional ones. PMID- 3048377 TI - Synovial chondromatosis of the temporomandibular joint. AB - Synovial chondromatosis of the temporomandibular joint is a rare, benign condition of metaplastic change of the synovium. A case is reported and the world literature is reviewed. PMID- 3048378 TI - Eosinophilic granuloma of the mandibular condyle. AB - An extremely rare location for eosinophilic granuloma of bone in the mandibular condyle has been described, illustrating the difficulties of diagnosis. Excision and reconstruction with a costochondral graft was successfully undertaken and remodelling of the graft occurred with the formation of a new condylar head. The patient is disease-free 17 months after surgery. The relevant literature is reviewed and suggestions made for management protocol. PMID- 3048376 TI - A randomized trial of progestogens in the primary treatment of endometrial carcinoma. PMID- 3048379 TI - Preoperative angiography in reconstructive microsurgery of the maxillofacial region. AB - The value of preoperative angiography of the donor and recipient sites in the reconstruction of the head and neck is reported. After neck-dissection an angiographic study of the carotid artery is routinely carried out as the vascular anatomy of the donor site can be determined only by angiography once surgery has been carried out. Angiography indicates not only the possible location for the planned microsurgical anastomosis, but also the potential difficulties which can arise from kinking and other changes in the vessel wall. The operative time can be reduced also by better preoperative planning of the best anastomotic sites. PMID- 3048380 TI - Bimaxillary proclination in patients of Afro-Caribbean origin. AB - The facial morphology of the Afro-Caribbean Negro is reviewed and the aims of treatment for bimaxillary proclination and its subsequent stability are discussed. Treatment approaches to various clinical situations are described, together with their known limitations. PMID- 3048381 TI - The emerging three-dimensional structure and function of 16S ribosomal RNA. PMID- 3048382 TI - Identification of a pterin derivative in Escherichia coli DNA photolyase. AB - DNA photolyase from Escherichia coli contains reduced flavin adenine dinucleotide plus a second chromophore, partially characterized in previous studies. Both chromophores function as sensitizers in catalysis. The second chromophore has been identified as a 6-substituted pterin derivative. The compound is oxidized with permanganate to yield 6-carboxypterin or reduced with sodium cyanoborohydride to yield a 5,6,7,8-tetrahydropterin derivative. The second chromophore exhibits spectral properties (lambda max = 360, 255 nm, pH 2) similar to that observed for 7,8-dihydropterin cations. The compound does not exhibit a spectrally detectable pKa around 4 but is converted to a dication (lambda max = 346, 255 nm) in strong acid (pKa approximately 1). Similar ionization behavior is observed with 7,8-dihydropterin derivatives that are alkylated at N(5). The instability of the second chromophore in weakly alkaline solution is due to a fully reversible conversion to a labile bleached form. As compared with other pterin derivatives, the hydrolytic instability is unusual but is very similar to that observed for 5,6-dialkyl-7,8-dihydropterinium salts. It is proposed that the second chromophore is a 7,8-dihydropterin with substituents at positions 5 and 6. The discovery that a pterin derivative functions as a photosensitizer in DNA repair is apparently the first example of a photobiological function for pterins. PMID- 3048383 TI - Inactivation of the Lactobacillus leichmannii ribonucleoside triphosphate reductase by 2'-chloro-2'-deoxyuridine 5'-triphosphate: stoichiometry of inactivation, site of inactivation, and mechanism of the protein chromophore formation. AB - The ribonucleoside triphosphate reductase (RTPR) of Lactobacillus leichmannii is inactivated by the substrate analogue 2'-chloro-2'-deoxyuridine 5'-triphosphate (ClUTP). Inactivation is due to alkylation by 2-methylene-3(2H)-furanone, a decomposition product of the enzymic product 3'-keto-2'-deoxyuridine triphosphate. The former has been unambiguously identified as 2 [(ethylthio)methyl]-3(2H)-furanone, an ethanethiol trapped adduct, which is identical by 1H NMR spectroscopy with material synthesized chemically. Subsequent to rapid inactivation, a slow process occurs that results in formation of a new protein-associated chromophore absorbing maximally near 320 nm. The terminal stages of the inactivation have now been investigated in detail. The alkylation and inactivation stoichiometries were studied as a function of the ratio of ClUTP to enzyme. At high enzyme concentrations (0.1 mM), 1 equiv of [5'-3H]ClUTP resulted in 0.9 equiv of 3H bound to protein and 83% inactivation. The amount of labeling of RTPR increased with increasing ClUTP concentration up to the maximum of approximately 4 labels/RTPR, yet the degree of inactivation did not increase proportionally. This suggests that (1) RTPR may be inactivated by alkylation of a single site and (2) decomposition of 3'-keto-dUTP is not necessarily enzyme catalyzed. The formation of the new protein chromophore was also monitored during inactivation and found to reach its full extent upon the first alkylation. Thus, out of four alkylation sites, only one appears capable of undergoing the subsequent reaction to form the new chromophore. While chromophore formation was prevented by NaBH4 treatment, the chromophore itself is resistant to reduction.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3048384 TI - Carbon-13 and deuterium isotope effects on the catalytic reactions of biotin carboxylase. AB - 13C and 2H kinetic isotope effects have been used to investigate the mechanism of enzymic biotin carboxylation. D(V/K) is 0.50 in 80% D2O at pD 8.0 for the forward reaction and 0.57 at pD 8.5 for the phosphorylation of ADP by carbamoyl phosphate. These values approach the theoretical maximum limit for a reaction in which a proton is transferred from a sulfhydryl to a nitrogen or oxygen base. Therefore, it appears that this portion of the reaction is at or near equilibrium. 13(V/K) at pH 8 is 1.007; the small magnitude of this number suggests that the reaction is almost fully committed by the time the carbon sensitive steps are reached. There does not appear to be a reverse commitment to the reaction under the conditions in which 13(V/K) was determined. A large forward commitment is consistent with the failure to observe positional isotope exchange from the beta gamma-bridge position to the beta-nonbridge position in [18O4]ATP or washout of 18O from the gamma-nonbridge positions. Transfer of 18O from bicarbonate to inorganic phosphate in the forward reaction was clearly observed, however. These observations suggest that biotin carboxylase exists in two distinct forms which differ in the protonation states of the two active-site bases, one of which is a sulfhydryl. Only when the sulfhydryl is ionized and the second base protonated can catalysis take place. Carboxylation of biotin is postulated to occur via a pathway in which carboxyphosphate in formed by nucleophilic attack of bicarbonate on ATP.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3048386 TI - Mammalian DNA polymerases alpha and delta: current status in DNA replication. PMID- 3048385 TI - Biosynthesis and molecular cloning of sulfated glycoprotein 1 secreted by rat Sertoli cells: sequence similarity with the 70-kilodalton precursor to sulfatide/GM1 activator. AB - Sulfated glycoprotein 1 (SGP-1) is one of the abundant proteins secreted by rat Sertoli cells. Pulse-chase labeling shows that SGP-1 is synthesized as a cotranslationally glycosylated 67-kilodalton (kDa) precursor which is posttranslationally modified to a 70-kDa form before secretion to the extracellular space. A plasmid cDNA library was constructed from immunopurified mRNA, and two overlapping clones coding for the entire protein coding sequence were isolated. The cDNAs represent 27 nucleotides of 5' noncoding sequence, 1554 nucleotides of coding sequence, and 594 nucleotides of 3' noncoding sequence. The derived SGP-1 sequence contains 554 amino acids and has a molecular weight of 61,123. Four potential N-glycosylation sites occur within the sequence. An internal region of SGP-1 shows 78% sequence identity with the 67 N-terminal amino acids described for human sulfatide/GM1 activator (SAP-1). Sequence comparisons suggest that SGP-1 is the precursor to sulfatide/GM1 activator; however, the secretion of the protein from Sertoli cells is distinct from the proteolytic processing and lysosomal compartmentalization which have been described for human fibroblasts. The presence of internal sequence similarity suggests that three additional binding sites may occur in SGP-1. Northern blots show similar levels of expression for the 2.6-kilobase SGP-1 mRNA in all tissues examined. The site of SGP-1 synthesis in testis was localized to Sertoli cells by immunofluorescence and in situ hybridization. PMID- 3048387 TI - Isolation, nucleotide sequence, and expression of a cDNA encoding pig citrate synthase. AB - Citrate synthase is a key enzyme of the Krebs tricarboxylic acid cycle and catalyzes the stereospecific synthesis of citrate from acetyl coenzyme A and oxalacetate. The amino acid sequence and three-dimensional structure of pig citrate synthase dimers are known, and regions of the enzyme involved in substrate binding and catalysis have been identified. A cloned complementary DNA sequence encoding pig citrate synthase has been isolated from a pig kidney lambda gt11 cDNA library after screening with a synthetic oligonucleotide probe. The complete nucleotide sequence of the 1.5-kilobase cDNA was determined. The coding region consists of 1395 base pairs and confirms the amino acid sequence of purified pig citrate synthase. The derived amino acid sequence of pig citrate synthase predicts the presence of a 27 amino acid N-terminal leader peptide whose sequence is consistent with the sequences of other mitochondrial signal peptides. A conserved amino acid sequence in the mitochondrial leader peptides of pig citrate synthase and yeast mitochondrial citrate synthase was identified. To express the pig citrate synthase cDNA in Escherichia coli, we employed the inducible T7 RNA polymerase/promoter double plasmid expression vectors pGP1-2 and pT7-7 [Tabor, S., & Richardson, C. C. (1985) Proc. Natl. Acad. Sci. U.S.A. 82, 1074-1078]. The pig citrate synthase cDNA was modified to delete the N-terminal leader sequence; then by use of a synthetic oligonucleotide linker, the modified cDNA was cloned into pT7-7 immediately following the initiator Met. A glutamate requiring (citrate synthase deficient), recA- E. coli mutant, DEK15, was transformed with pGP1-2 and then pT7-7PCS. pT7-7PCS complemented the E. coli gltA mutation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3048388 TI - Complementary oligodeoxynucleotide probes of RNA conformation within the Escherichia coli small ribosomal subunit. AB - The large RNA molecule within each ribosomal subunit is folded in a specific and compact form. The availability of specific 16S RNA sequences on the surface of the small ribosomal subunit has been probed by using complementary oligodeoxynucleotides. The hybridization of 8-15-nucleotide-long oligomers to their RNA complements within the subunit was quantitated by using a nitrocellulose membrane filter binding assay. The probes have been grouped into classes on the basis of sequence-specific binding ability under different conditions of ionic environment, incubation temperature, and subunit activation state [as defined by the ability to bind phenylalanyl-tRNA in response to a poly(uridylic acid) message]. Oligodeoxynucleotides complementary to nucleotides flanking 7-methylguanosine residue 527 and to the 3'-terminal sequence bound 30S subunits regardless of the activation state. Oligodeoxynucleotides that complement 16S ribosomal RNA residues 1-16, 60-70, 685-696, and 1330-1339 and the sequence adjacent to the colicin E3 cleavage site at residue 1502 all bound efficiently only to subunits in an inactivated conformation. Probes complementary to residues 1-11 and 446-455 bound only inactivated subunits, and then with low efficiency. Sequences complementary to nucleotides 6-16, 99-109, 1273-1281, and 1373-1383 bound 30S subunits poorly regardless of the activation state. With one exception, each probe was bound by native or heat-denatured 16S ribosomal RNA (as determined by size-exclusion chromatography). We conclude that complementary oligodeoxynucleotide binding efficiency is a sensitive measure of the availability of specific RNA sequences under easily definable conditions. PMID- 3048389 TI - Spectroscopic and hydrodynamic studies reveal structural differences in normal and transforming H-ras gene products. AB - We have recorded the circular dichroism spectra of the cellular and the viral H ras gene products both in the absence and in the presence of guanine nucleotides and analyzed these spectra in terms of the secondary structure composition of these proteins. It is shown that the GTP complex of the ras proteins has a different secondary structure composition than the GDP complex and, furthermore, that there are differences in the secondary structure of the viral ras protein and the cellular ras protein. We have also recorded and analyzed the circular dichroism spectrum of the isolated guanine nucleotide binding domain of the Escherichia coli elongation factor Tu (EF-Tu), which has been considered as a model for the tertiary structure of the ras proteins [McCormick, F., Clark, B. F. C., LaCour, T. F. M., Kjeldgaard, M., Norskov-Lauritsen, L., & Nyborg, J. (1985) Science (Washington, D.C.) 230, 78-82]. Our data show that the guanine nucleotide binding domain of EF-Tu (30% alpha-helix and 16% beta-pleated sheet for the GDP complex) has quite a different secondary structure composition than the ras proteins (e.g., the cellular ras protein has 47% alpha-helix and 22% beta-pleated sheet for the GDP complex), indicating that the protein core comprising the guanine nucleotide binding site might be similar but that major structural differences must exist at the portion outside this core. Normal and transforming ras proteins also differ slightly in their hydrodynamic properties as shown by sedimentation velocity runs in the analytical ultracentrifuge.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3048391 TI - Isotope partitioning in the adenosine 3',5'-monophosphate dependent protein kinase reaction indicates a steady-state random kinetic mechanism. AB - Isotope partitioning beginning with the binary E.MgATP and E.N-acetyl-Leu-Arg-Arg Ala-Ser-Leu-Gly (Ser-peptide) complexes indicates that the kinetic mechanism for the adenosine 3',5'-monophosphate dependent protein kinase is steady-state random. A total of 100% of the initial radioactive E.MgATP complex is trapped as phospho-Ser-peptide at infinite Ser-peptide concentration at both low and high concentration of uncomplexed Mg2+, suggesting that the off-rate of MgATP from the E.MgATP.Ser-peptide complex is slow relative to the catalytic steps. Km for Ser peptide in the trapping reaction decreases from 17 microM at low Mg2+ to 2 microM at high Mg2+, indicating that Mg2+ decreases the off-rate for MgATP from the E.MgATP complex. A total of 100% of the radioactive E.Ser-peptide complex is trapped as phospho-Ser-peptide at low Mg2+, but only 40% is trapped at high Mg2+ in the presence of an infinite concentration of MgATP, suggesting that the off rate for Ser-peptide from the central complex is much less than catalysis at low but not at high Mg2+. In support of this finding, the Ki for Leu-Arg-Arg-Ala-Ala Leu-Gly (Ala-peptide) increases from 0.27 mM at low Mg2+ to 2.4 mM at high Mg2+. No trapping was observed at either high or low Mg2+ for the E.MgADP complex up to a phospho-Ser-peptide concentration of 5 mM. Thus, it is likely that in the slow reaction direction the kinetic mechanism is rapid equilibrium.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3048390 TI - Stereochemistry of phospho group transfer catalyzed by a mutant alkaline phosphatase. AB - The stereochemical course of the phospho group transfer catalyzed by mutant (S102C) alkaline phosphatase from Escherichia coli was investigated by using 31P nuclear magnetic resonance spectroscopy. Transphosphorylation from 4-nitrophenyl (Rp)-[16O, 17O, 18O]phosphate to (S)-propane-1,2-diol occurs with overall retention of configuration at phosphorus. This result is consistent with the view that the hydrolysis of substrates by this mutant enzyme proceeds by way of a covalent phosphoenzyme intermediate in the same manner as the wild-type alkaline phosphatase. PMID- 3048392 TI - Ligand-induced structural constraints in human dihydrofolate reductase revealed by peptide-specific antibodies. AB - Peptides from human dihydrofolate reductase (DHFR) generated by cyanogen bromide cleavage and corresponding to residues 15-52, 53-111, 112-125, and 140-186 (carboxyl terminus) were purified and used to immunize rats. Titration of the immune sera against denatured human DHFR by solid-phase immunoassay showed that peptides 15-52 and 140-186 were relatively highly immunogenic, unlike the native enzyme which is most immunogenic in the sequence 53-111. The antisera were specific for the corresponding peptides used for immunization. Antibodies to peptides 15-52, 53-111, and 140-186 cross-reacted with native human DHFR in solution in competition assays. However, the binding of nicotinamide adenine dinucleotide phosphate (reduced) (NADPH) and the inhibitors folate and methotrexate, both in binary and in ternary complexes with the enzyme, caused a striking reduction in binding of antibody. Using a sensitive radioactive assay, it was found that antisera to peptides 15-52 and 140-186, both of which exhibited a high antibody titer, caused significant inhibition of DHFR. Because peptide 140 186 does not include any active-site residues, it is concluded that at least in this case all the antibodies bound to regions outside the active site. Since comparison of the X-ray structures of the chicken liver DHFR holoenzyme with the apoenzyme reveals no changes in secondary structural elements (alpha-helices and beta-sheets), the reduction in antibody binding to DHFR-ligand complexes must not involve epitopes within these structures.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3048393 TI - Reoxidation of the class I disulfides of the rat adipocyte insulin receptor is dependent upon the presence of insulin: the class I disulfide of the insulin receptor is extracellular. AB - Elements of the quaternary structure of the native and dithiothreitol- (DTT) reduced rat adipocyte insulin receptor have been elucidated by vectorial probing and subunit cross-linking. The charged reducing agents glutathione and beta mercaptoethylamine were used to reduce the class I disulfides of the receptor in intact adipocytes, demonstrating the extracellular location of the disulfide directly. This interpretation was confirmed by use of DTT as a reducing agent and the nonpermeant sulfhydryl blocking reagent Thiolyte MQ to prevent the reoxidation of the class I sulfhydryl groups which occurred when they were not blocked. It was found that the above reoxidation of the receptor is dependent on the concentration of insulin in the nanomolar range, not occurring measurably at 4 degrees C in its absence. Cross-linking studies with ethylene glycol bis(succinimidyl succinate) demonstrated that the alpha subunits could not be cross-linked to each other after reduction of the class I disulfides, suggesting that the interaction between the receptor heterodimers may be due primarily to the disulfide bonds. PMID- 3048394 TI - Direct photoaffinity labeling of ribonucleotide reductase from Escherichia coli using dTTP: characterization of the photoproducts. AB - Subunit B1 of Escherichia coli ribonucleotide reductase contains one type of allosteric binding site that controls the substrate specificity of the enzyme. This site binds the allosteric effector dTTP as well as other nucleoside triphosphates. Cross-linking of dTTP to protein B1 by direct photoaffinity labeling, as well as the isolation and sequence determination of the labeled tryptic peptide, has recently been reported [Eriksson, S., Sjoberg, B.-M., Jornwall, H., & Carlquist, M. (1986) J. Biol. Chem. 261, 1878-1882]. In this study, we have further purified the dTTP-labeled peptide and characterized it using UV spectroscopy. Two types of dTTP-cross-linked peptide were found: one having an absorbance maximum at 261 nm typical for a dTTP spectrum, i.e., containing an intact 5,6 double bond, and one minor form with low absorbance at 261 nm. In both cases, the same amino acid composition was found, corresponding to the peptide Ser291-X-Ser-Gln-Gly-Gly-Val-Arg299 in the B1 sequence with X being Cys-292 cross-linked to dTTP. Isotope labeling experiments revealed that one proton in the 5-methyl group of thymine was lost during photoincorporation. Therefore, the cross-linking occurs via the 5-methyl group to Cys-292 in a majority of incorporated dTTPs, but a second, possibly 5,6-saturated form of incorporated nucleotide was also detected. The reasons for the high stereospecificity of the reaction and the possible structure of the allosteric site of protein B1 are discussed. PMID- 3048396 TI - Isolation of multiple forms of DNA polymerase delta: evidence of proteolytic modification during isolation. AB - The subunit structures of a number of human placenta DNA polymerase delta preparations were investigated by Western blotting with polyclonal antisera and by activity staining following polyacrylamide gel electrophoresis. When immunoblots and activity stains were performed on different enzyme preparations, putative catalytic subunits of (a) 170, (b) 120, or (c) 50-70 kilodaltons (kDa) were observed. It was also observed that the lower molecular weight forms could be generated upon storage of the preparations. Western blotting of human placental tissue extracts showed that the major immunoreactive polypeptide was 160-170 kDa. Treatment of the extracts with trypsin or Staphylococcus aureus V8 protease led to the generation of immunoreactive polypeptides of lower molecular weights. These studies suggest that the 120-kDa and lower forms of the enzyme are generated via uncontrolled proteolysis and provide a rationale for the observation of different apparent subunit structures previously reported for DNA polymerase delta. In addition, these findings suggest that DNA polymerase delta has a catalytic domain which resides in a protease-resistant domain. PMID- 3048395 TI - Structural comparison between oxidized and reduced Escherichia coli thioredoxin. Proton NMR and CD studies. AB - Escherichia coli thioredoxin (Mr 11,700) usually functions as a hydrogen carrier protein that undergoes reversible oxidation/reduction reactions of its active site disulfide linkage. By use of a number of assigned and identified resonances in one- and two-dimensional 1H NMR spectra, the two forms of the protein have been compared. Only groups that are relatively close to the active-site Cys-32, Cys-35 linkage such as Trp-28, Trp-31, Phe-27, Ala-29, and Val-25 undergo substantial changes in their 1H NMR chemical shift upon reduction. Various residues that are further removed from the active site, like Tyr-49, Tyr-70, His 6, Phe-12, Phe-81, and Phe-102, appear to be little affected (less than 0.02 ppm) by the reduction, suggesting that the rest of the protein structure is not much affected. Thus, the structural changes that occur upon reduction appear to be localized to the disulfide-containing turn and the central strand of the twisted beta-sheet that directly leads to this turn. Notwithstanding the apparent similarity in the secondary and tertiary structures of the oxidized and reduced forms of the protein, the thermal stability of the protein decreases by 10 degrees C upon the reduction of the single disulfide. This was found by both 1H NMR and near- and far-ultraviolet circular dichroism studies. Oxidized thioredoxin was also more resistant to alkaline denaturation. Furthermore, the exchange rate of the relatively stable slow-exchanging backbone amide protons that are part of the core of the twisted five-stranded beta-sheet of thioredoxin increases substantially after reduction.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3048397 TI - The dimer of the beta subunit of Escherichia coli DNA polymerase III holoenzyme is dissociated into monomers upon binding magnesium(II). AB - The beta subunit of Escherichia coli DNA polymerase III holoenzyme binds Mg2+. Reacting beta with fluoresceinmaleimide (FM) resulted in one label per beta monomer with full retention of activity. Titration of FM-beta with Mg2+ resulted in a saturable 11% fluorescence enhancement. Analysis indicated that there was one noncooperative magnesium binding site per beta monomer with a dissociation constant of 1.7 mM. Saturable fluorescence enhancement was also observed when titration was with Ca2+ or spermidine(3+) but not with the monovalent cations Na+ and K+. The Mg2+-induced fluorescence enhancement was specific for FM-beta and was not observed with FM-glutathione, dimethoxystilbenemaleimide-beta, or pyrenylmaleimide-beta. Gel filtration studies indicated that the beta dimer monomer dissociation occurred at physiologically significant beta concentrations and that the presence of 10 mM Mg2+ shifted the dimer-monomer equilibrium to favor monomers. Both the gel-filtered dimers and the gel-filtered monomers were active in the replication assay. These and other results suggested that the fluorescence increase which accompanies beta dissociation is due to a relief from homoquenching of FM when the beta dimer dissociates into monomers. PMID- 3048399 TI - Multiantennary group-specific polysaccharide of group B Streptococcus. AB - The group-specific antigen of group B Streptococcus is composed of four different oligosaccharide units of Mw 766 (III), 1277 (II), 1462 (IV), and 1788 (I). The major constituent sugars of the oligosaccharides are alpha-L-rhamnopyranose, alpha-D-galactopyranose, 2-acetamido-2-deoxy-beta-D-glucopyranosyl, and D glucitol except that III does not contain alpha-D-galactopyranosyl or 2-acetamido 2-deoxy-beta-D-glucopyranosyl residues and IV contains no D-glucitol but has one additional beta-L-rhamnopyranosyl residue. The structures of II and III have been previously elucidated [Michon, F., Katzenellenbogen, E., Kasper, D. L., & Jennings, H. J. (1987) Biochemistry 26, 476-486]. In the group B antigen all the oligosaccharides are linked by one type of phosphodiester bond from O6 of the D glucitol residue of one oligosaccharide to O6 of the alpha-D-galactopyranosyl residue of the next to form a complex and highly branched multiantennary structure. However, despite the heterogeneous nature of its component oligosaccharides, some order has been identified in the biosynthesis of the group B antigen from chemical and enzymatic sequence studies. Because III lacks an alpha-D-galactopyranosyl residue but has a D-glucitol residue, it is situated at the reducing terminus of all the branches of the group B antigen where it is always adjacent to a II moiety. Conversely, IV has an alpha-D-galactopyranosyl residue but has no D-glucitol and is therefore located at the reducing terminus of the group B antigen where it probably functions as a linker molecule between the group B polysaccharide and the cell wall peptidoglycan of the group B streptococcal organisms. Oligosaccharide I contains two alpha-D-galactopyranosyl residues and one D-glucitol residue and thus constitutes the branch point in the group B antigen, whereas II contains one of each of the above residues and therefore is situated in linear interchain positions. The group B antigen is highly branched and probably has a unique multiantennary structure. PMID- 3048398 TI - Identification of carbohydrate binding protein 35 in heterogeneous nuclear ribonucleoprotein complex. AB - In previous studies, a lectin designated as carbohydrate binding protein 35 (CBP35) was identified in the nucleus and cytoplasm of cultured mouse 3T3 fibroblasts. In the present study, we observed that treatment of Triton X-100 permeabilized 3T3 cells with ribonuclease A released CBP35 from the nuclei, while parallel treatment with deoxyribonuclease I failed to do so. This conclusion was based on (a) immunofluorescence analysis of the nuclear residue after detergent and enzymatic treatments and (b) immunoblotting analysis of the supernatant fraction produced by these treatments. These results indicate that CBP35 may be associated with the ribonucleoprotein elements of the 3T3 cell nuclei. In corroboration with this conclusion, fractionation of the nucleoplasm derived from 3T3 cells on a cesium sulfate gradient (1.25-1.75 g/mL) localized CBP35 in fractions with densities of 1.30-1.32 g/mL, corresponding to the range of densities reported for heterogeneous nuclear ribonucleoprotein complex (hnRNP). Conversely, when nucleoplasm was fractionated on an affinity column of Sepharose derivatized with N-(epsilon-aminocaproyl)-D-galactosamine, the bound and eluted fraction contained RNA, as well as a set of polypeptides whose molecular weights matched those reported for the core particle of hnRNP. One of these polypeptides was identified as CBP35. These results suggest that CBP35 is a component of hnRNP. PMID- 3048400 TI - Old and new concepts of the membrane transport for glucose in cells. PMID- 3048401 TI - Nucleoside and nucleobase transport in animal cells. PMID- 3048402 TI - Solute translocation across the mammalian lysosome membrane. PMID- 3048403 TI - Enzymes II of the phosphoenolpyruvate-dependent sugar transport systems: a review of their structure and mechanism of sugar transport. PMID- 3048404 TI - The biology and biochemistry of the glucose transporter. PMID- 3048405 TI - The role of sequence in the stabilization of left-handed DNA helices in vitro and in vivo. PMID- 3048406 TI - Inhibition of DNA-dependent RNA synthesis by 8-methoxypsoralen. AB - The effect of the photobinding of 8-methoxypsoralen to phage T7 DNA on different steps of RNA synthesis in vitro was assayed. Total RNA synthesis is reduced to a few percent and the transcript size is decreased, as shown by means of gel filtration on a Sepharose 4B column when DNA of the adduct content of six drug molecules per 10(3) nucleotides is used. The initiation of RNA chains seems to be less affected, as inferred from an abortive initiation assay. Synthesis of pppApU on DNA of the same adduct content is inhibited to 34% of the corresponding controls, while the overall RNA synthesis is inhibited to 6%. The amount of the enzyme needed for maximal retention of DNA, the kinetics of its binding and the decay of the polymerase-DNA complex at high ionic strength (or on decrease of the temperature) are similar with DNA either irradiated in the absence of the drug or DNA bearing six 8-methoxypsoralen molecules per 10(3) nucleotides. It is concluded from this study that 8-methoxypsoralen partially inhibits initiation and blocks movement of RNA polymerase along the template, inducing premature termination. It does not appear to influence the binding of the enzyme to DNA. PMID- 3048407 TI - Spectroscopic analysis of DNA base-pair opening by Escherichia coli RNA polymerase. Temperature and ionic strength effects. AB - The interaction of Escherichia coli RNA polymerase with poly[d(A-T)] and poly[d (I-C)] was studied by difference absorption spectroscopy at temperatures, from 5 to 45 degrees C in the absence and presence of Mg2+. The effect of KCl concentration, at a fixed temperature, was studied from 12.5 to 400 mM. Difference absorption experiments permitted calculation of the extent of DNA opening induced by RNA polymerase and estimation of the equilibrium constant associated with the isomerization from a closed to an open RNA polymerase-DNA complex. delta H0 and delta S0 for the closed-to-open transition with poly[d(A T)] or poly[d(I-C)] complexed with RNA polymerase are significantly lower than the values associated with the helix-to-coil transition for the free polynucleotides. For the RNA polymerase complexes with poly[d(A-T)] and poly[d(I C)] in 50 mM KCl, delta H0 approximately 15-16 kcal/mol (63-67 kJ/mol) and delta S0 approximately 50-57 cal/K per mol (209-239 J/K per mol). The presence of Mg2+ does not change these parameters appreciably for the RNA polymerase-poly[d(A-T)] complex, but for the RNA polymerase-poly[d(I-C)] complex in the presence of Mg2+, the delta H0 and delta S0 values are larger and temperature-dependent, with delta H0 approximately 22 kcal/mol (92 kJ/mol) and delta S0 approximately 72 cal/K per mol (approx. 300 J/K per mol) at 25 degrees C, and delta Cp0 approximately 2 kcal/K per mol (approx. 8.3 kJ/K per mol). The circular dichroism (CD) changes observed for helix opening induced by RNA polymerase are qualitatively consistent with the thermally induced changes observed for the free polynucleotides, supporting the difference absorption method. The salt-dependent studies indicate that two monovalent cations are released upon helix opening. For poly[d(A-T)], the temperature-dependence of enzyme activity correlates well with the helix opening, implying this step to be the rate-determining step. In the case of poly[d(I-C)], the same is not true, and so the rate-determining step must be a process subsequent to helix opening. PMID- 3048408 TI - The expression of functional ricin B-chain in Saccharomyces cerevisiae. AB - Yeast cells transformed with plasmids containing ricin B-chain coding sequences expressed this heterologous protein. When ricin B-chain was expressed in a form which resulted in its deposition in the yeast cytosol it formed insoluble aggregates which were devoid of galactose-binding activity. In contrast, when DNA fusions were constructed, in which the B-chain coding sequence was preceded by either the preproalpha-factor leader sequence or the native preproricin signal sequence, the recombinant B-chain products were soluble and biologically active. Both the homologous yeast signal peptide and the heterologous plant signal peptide directed the expressed product into the lumen of the yeast endoplasmic reticulum. As a result, the recombinant B-chain products were processed at the N terminus, glycosylated and folded into an active conformation, presumably stabilized by correct intrachain disulphide bond formation. PMID- 3048410 TI - Increased synthesis of human parathyroid hormone in Escherichia coli through alterations of the 5' untranslated region. AB - The expression of human parathyroid hormone (hPTH) in Escherichia coli was optimized by variations of the spacing sequence between the ribosome-binding site (RBS) and the beginning of the gene (ATG) and by increasing the complementarity of the RBS to the 16 S rRNA. The expression level of 3 micrograms/liter increased more than 100-fold to 475 micrograms/liter as a direct consequence of modifications in the region 5' of the gene. PMID- 3048409 TI - The phosphoprotein p53 is down-regulated post-transcriptionally during embryogenesis in vertebrates. AB - The phosphoprotein p53 has been investigated mainly because of its relationship with tumorigenic transformation. In this communication, we report that, during the embryonal development of mouse and chicken, there is a decline in the steady state levels of the p53 protein and an equal decline in p53 mRNA. During the development of the chicken, the relative rates of p53 transcription appear to be constant. p53 mRNA is relatively stable (half-life greater than 12 h) in both chicken and mouse embryos. We conclude that (i) the down-modulation of p53 mRNA (and of protein) during embryonal development has been well conserved during the evolution of the vertebrate, implying that the p53 protein may have a function in embryonal development; and (ii) the mechanism of control is apparently mainly on a post-transcriptional level. PMID- 3048411 TI - The allosteric effect of salt on human mast cell tryptase. AB - The inhibitory effect of potassium chloride and ammonium sulphate on purified human skin tryptase and bovine trypsin was studied enzyme-kinetically, using Z Gly-Pro-Arg-pNA, Z-Gly-Pro-Arg-AMC, benzoyl-L-arginine ethyl ester (BAEE) and tosyl-L-arginine methyl ester (TAME) as substrates. With increasing salt concentrations, the curve of reaction velocity vs. substrate concentration changed from hyperbolic to sigmoidal when anilide substrates (Z-Gly-Pro-Arg-pNA or -AMC) were used. Only the Km value increased, while the Vmax value remained unchanged. The trend was similar with BAEE or TAME as the substrates. However, the effect of salt on the hydrolysis of these ester substrates was not as strong as on the hydrolysis of anilide substrates, and sigmoidal kinetics were not observed even at the highest KCl concentration (0.7 M) used. Heparin, used as a stabilizer, had no influence on this phenomenon, but it did slightly decrease the apparent Km and Vmax values in low-salt conditions. By comparison, trypsin was not as strongly affected by salt as tryptase, and the inhibition type was mixed competitive and non-competitive. The present results indicate that the salt acts on tryptase as an allosteric effector, and this should be carefully considered when enzyme kinetic parameters and enzyme activity of skin tryptase are measured. PMID- 3048412 TI - Porcine muscle prolyl endopeptidase: limited proteolysis of tryptic peptides from hemoglobin beta-chains at prolyl and alanyl bonds. AB - Tryptic peptides from hemoglobin (Hb) beta-chains were used as model substrates for limited proteolysis by prolyl endopeptidase (EC 3.4.21.26) from porcine muscle. From the physicochemical and enzymatic properties of prolyl endopeptidase the conditions for routine digestion were established as follows: the molar ratio of enzyme to substrate was 1 to 100, and the reaction was carried out in sodium phosphate buffer (pH 6.4) at 37 degrees C for 4 h. Under these conditions the peptide bonds on the carboxyl terminal sides of proline and alanine residues in the tryptic peptides from Hb beta-chains (with Mr values of less than 2100) were hydrolyzed by the enzyme with the exception of the amino terminal alanyl bond and aminoacyl alanyl bond. In addition, one of five seryl bonds was cleaved by the enzyme. However, the Hb beta-chain itself, Mr 16,600, and its two CNBr-peptides with Mr 10,200 and Mr 6400, respectively, were not hydrolyzed. Under the same conditions a prolyl bond in oxidized B-chains of insulin, Mr 3400, was partially digested, and an alanyl bond was not hydrolyzed. The data indicate that the prolyl endopeptidase is useful for the limited proteolysis of peptides with relative masses of less than 3000 at both prolyl and alanyl bonds. PMID- 3048414 TI - Immunogenic properties of mannose-containing ceramide tetrasaccharide from fresh water bivalve, Hyriopsis schlegelii. AB - Antiserum against GlcNAc beta 1----2Man alpha 1----3Man beta 1----4Glc beta 1--- 1Cer (MlOse4Cer), a mannolipid isolated from spermatozoa of the fresh-water bivalve Hyriopsis schlegelii, was elicited in rabbits by repeated injection of a mixture of hapten-bovine serum albumin with Freund's adjuvant. The specificity of the affinity-purified antibody obtained from the serum was based on two forms of enzyme-immunodetection of its binding to structurally related glycolipids, either adsorbed to microtiter plates or chromatographed on thin-layer plates. The purified antibody exhibited a significant cross-reactivity with GlcNAc beta 1--- 2Man alpha 1----3(Xyl beta 1----2)Man beta 1----4Glc beta 1----1Cer, (MIXOse5Cer) containing a core structure closely related to MlOse4Cer, but almost unrelated to other glycolipids. Distribution of MlOse4Cer and MlXOse5Cer in various bivalve and snail glycolipid extracts were screened in thin-layer immunobinding assays by using this purified specific antibody. The presence of the glycolipid antigens was limited to certain taxonomic orders of shellfish species. PMID- 3048413 TI - Catalysis of proline isomerization during protein-folding reactions. AB - The enzyme peptidylprolyl cis-trans isomerase (PPI) is known to catalyze proline isomerization in short proline-containing peptides. If PPI can be shown to generally catalyze isomerization of proline residues in proteins, then it would be a valuable diagnostic reagent for recognition of isomerization, which has proven to be extremely difficult to characterize by other methods. In this study, the catalytic effect of PPI on the slow refolding reactions of seven different proteins has been studied, and in only two cases (RNase T1 and cytochrome c) could significant catalysis be seen. PPI also caused no enhancement in the rate for the 'subtle' conformational changes of native concanavalin A or native Fragment I of prothrombin, which have been suggested to be rate-limited by proline isomerization. There was a small effect of PPI observed for the generation of native RNAase A from the fully-reduced form when the glutathione concentration was low. The conclusion from these studies is that PPI can weakly catalyze some protein processes which are rate-limited by proline isomerization, but probably exhibits no measureable catalysis toward others. This somewhat limits the usefulness of PPI as a diagnostic reagent for proline isomerization. PMID- 3048415 TI - Gangliosides in the blood plasma: levels of ganglio-series gangliosides in the plasma after administration of brain gangliosides. AB - The temporal change in the levels of the gangliotetraose-series gangliosides, i.e., GMla, GDla, GD1b, GT1b, in the blood plasma after intramuscular administration of bovine brain gangliosides (5 mg/kg) to beagle dogs (11.3-12.2 kg) was determined with high sensitivity by a recently developed thin-layer chromatography/enzyme-immunostaining method (Hirabayashi, Y., Koketsu, K., Higashi, H., Suzuki, Y., Matsumoto, M., Sugimoto, M. and Ogawa, T. (1986) Biochim. Biophys. Acta 876, 178-182). The amounts of GMla, GDla, GD1b, GT1b and their combined total in the plasma of beagle dogs before administration of gangliosides were 21 +/- 1, 36 +/- 7, 15 +/- 2, 16 +/- 2 and 88 +/- 6 pmol/ml of blood plasma, respectively. Trapezoidal calculation showed that the times of the maximum levels of GMla, GDla, GDlb, GTlb and the total of the their levels in the plasma were 8.0 +/- 1.2, 8.7 +/- 0.7, 6.3 +/- 2.0, 17.0 +/- 7.0 and 8.7 +/- 0.7 h after the administration of gangliosides, and their maximum concentrations were 517 +/- 37, 654 +/- 53, 160 +/- 5, 184 +/- 20 and 1383 +/- 74 pmol/ml, respectively. The maximum level of each ganglioside decreased gradually, reaching the normal level after 10 days. The half-maximum level of each ganglioside occurred 2-3 days after the administration. Asialo GM1 (GA1) was not detected plasma at any of the test times. PMID- 3048416 TI - Identity of soluble thiamin-binding protein with thiamin-repressible acid phosphatase in Saccharomyces cerevisiae. AB - Two secretory glycoproteins of Saccharomyces cerevisiae, a soluble thiamin binding protein and a thiamin-repressible acid phosphatase, were shown to be repressed to a similar extent by excess thiamin in the growth medium. Thiamin repressible acid phosphatase was co-purified throughout the purification of the soluble thiamin-binding protein. Purified and deglycosylated soluble thiamin binding proteins exhibited both thiamin-binding and acid phosphatase activity on non-denaturing polyacrylamide gel electrophoresis. Heat treatment of the purified soluble thiamin-binding protein caused a decrease in both activities with a similar inactivation profile. Furthermore, two thiamin-repressible acid phosphatase-defective mutants isolated had no and decreased soluble thiamin binding activity, respectively. From the results, it was concluded that the soluble thiamin-binding protein is identical to the thiamin-repressible acid phosphatase in S. cerevisiae. PMID- 3048417 TI - Effective stage in the cell cycle for control of the budding direction of cdc mutants of Saccharomyces cerevisiae using electric stimulus. AB - Cell division cycle (cdc) mutants of Saccharomyces cerevisiae were used to determine the most effective stage for the directional control of cell budding using an electric stimulus. The selected mutants were cdc 35 and cdc 28, which could be reversibly arrested before spindle pole body satellite formation (SPBSF) and spindle pole body duplication (SPBD), respectively. The budding direction (theta) was defined so that the direction parallel to that of the electric field was 0 degree. Considering the symmetry of the experimental conditions, the range of theta was defined as 0-90 degrees. The electric stimulus applied in the present study was alternating pulses (pulse height, +/- 15 V; pulse width at half pulse height, 5 microseconds; frequency; 10 kHz). The peak height of the cross membrane potential was estimated as 472 mV, which was sufficient to induce considerable strain in the cell membrane. In the case of cdc 35, the 95% confidence interval (95% CI) of the budding direction was 7-25 degrees when subjected to electric stimulus, while the 95% CI of the budding direction without electric stimulus was 35-57 degrees. In the case of cdc 28, 95% CI values of the budding direction with and without electric stimulus were 1229 degrees and 23-56 degrees, respectively. These results demonstrate that the stage after SPBD is effective for the directional control of yeast cell budding using an electric stimulus. Simultaneously, an electric stimulus reduced the cell budding time of both the cdc mutants used. Therefore, the electric stimulus was also effective in promoting cell cycle progression under the present conditions. PMID- 3048419 TI - [Changes in the volutin levels in yeast cells during cell cycle]. AB - Dimeric distribution of the cell number of Saccharomyces cerevisiae is analysed according to the volume and content of volutin in them. Dynamics of the age composition of the population in time when changing the medium limiting growth for the balanced one is considered. The data obtained indicate that volutin may serve as a regulator when the yeast cells pass the starting point in the division cycle, however, this event has no contrary effect on volutin accumulation. PMID- 3048418 TI - Characterization of vesicles containing insulin-responsive intracellular glucose transporters isolated from 3T3-L1 adipocytes by an improved procedure. AB - Our previously described immunoadsorption method for the isolation of vesicles containing the insulin-responsive intracellular glucose transporters from 3T3-L1 adipocytes has been improved in two ways. First, the minimal number of g minutes required to sediment the plasma membranes from the cell homogenate has been determined and, as a result, the supernatant used for immunoadsorption in the new procedure contained twice as much of the intracellular transporters. Second, the immunoadsorption has been performed with affinity-purified antibodies directed against the carboxy terminal peptide of the transporter, rather than against the entire protein. 10(7) cells (10 mg protein) yielded about 12 micrograms of vesicular protein and 11 micrograms of vesicular phospholipid. The transporter constituted 3% of the protein in the vesicles; this amount equates to approx. eight copies of the transporter per 50 nm vesicle. The polypeptide composition of the vesicles was determined by gel electrophoresis and protein staining. Major components, other than the glucose transporter, are polypeptides of Mr 270,000, 245,000, 165,000 and 115,000. The vesicles contained several phosphoproteins; the major ones have a Mr of 245,000, 190,000, 115,000 and 25,000. Insulin treatment of adipocytes did not significantly change the phosphoprotein composition of the vesicles. The vesicles were not enriched in the Golgi marker enzyme, galactosyltransferase. The cellular content of the marker for the trans-Golgi reticulum, sialyltransferase, was too low to detect. PMID- 3048420 TI - [Lipopolysaccharide-induced hydrolysis of plasmalogenic phosphatidylethanolamine in human platelets]. AB - The composition of human platelet major phospholipids-phosphatidylcholine (PC), phosphatidylinositol (PI), phosphatidylserine (PS), phosphatidic acid (PA), sphingomyelin (SM), plasmalogenic and diacyl species of phosphatidylethanolamine (PPE and APE, respectively) was quantitatively analyzed by high performance liquid chromatography. Incubation (10 min, 37 degrees C) of washed platelets with lipopolysaccharide B (LPS) of Salmonella typhimurium was found to produce (in the absence of aggregation) marked hydrolysis of PI (ca. 15%) and PPE (ca. 19%) containing the bulk of polyenic fatty acids. PC and APE were less degraded (8 9%), while the amounts of PS and SM were practically unchanged and the level of PA rose by 20%. Addition of thrombin to LPS-pretreated platelets resulted in their more rapid aggregation which was accompanied by a decreased and nearly equal hydrolysis of APE and PPE (7-8%) as compared with control platelets (10 and 12%, respectively). The extent to which PI was degraded (ca. 34%), by the action of thrombin was not affected by preliminary incubation with LPS. It is suggested that thrombin (as well as LPS) activating endogenous phospholipase(s) A2 can liberate from PPE not only arachidonic acid but also other essential polyenic fatty acids present in PPE in relatively high amounts. Besides, the agents studied may activate the intrinsic platelet system of rapid arachidonoyl transfer from diacyl PC and PE to PPE. PMID- 3048421 TI - [The respiratory center--the regulator of the respiratory system]. AB - The authors consider the respiratory centre to be the regulator of the respiratory system and to consist of 3 main functional blocks: chemoregulator, respiratory rhythm autogenerator and mechanoregulator, functions of which are provided by the neurons of medulla oblongata. The main aim of chemoregulator block is to maintain the level of ventilation volume speed, which is necessary to compensate the difference between the signals of setting and the firing from the chemoreceptors. The main aim of mechanoregulator block is to provide the functioning of the regulation loop of the respiratory muscles comparing the signals which come from the respiratory autogenerator, and the firing of the mechanoreceptors. The generator unit of the respiratory centre is a set of rhythm forming associations, the system of 4 neurons (early and late inspiratory and expiratory) are typical among them. The neurons are connected by recurrent inhibitory bonds: the neuron of each rhythm-forming group, successively becoming excited, inhibits the two preceding neurons in the cycle; for all this the neuron of the successive group is released from inhibition and in such a way the rhythmogenesis occurs. The respiratory centre forms a common unit for chemo- and mechanoreceptor loops, through which the circuits of feedback for both loops are connected, providing the regulation of breathing. PMID- 3048422 TI - [Genetic demographic approach in anthropological research. IV. The information value of genealogical tables and the level of inbreeding in Khakass populations]. AB - The average volume of genealogical information in two Khakass ethnographic groups -Kizil and Sagai populations was 0.451 and 0.331 taking into account 7 and 10 generations under study correspondingly. The inbreeding coefficients calculated with the help of genealogical method are 0.0012 and 0.0025 and using the group of potentially or conditionally consanguineous marriages--0.0051 and 0.0108. The common inbreeding coefficient (FIT) and relative impact into it of the random (FST) and nonrandom (FIS) components received by means of the isonymy method in Kizil ethnographic group were 0.0121, 0.026 and -0.0143, while in the Sagai group the figures were 0.020, 0.042 and -0.023. The collection and analysis of the genealogical information presented non-graphically gives wide opportunities in description of the population structures as well in a number of allied branches of science where the genealogical (graphic) method is used. PMID- 3048423 TI - [Dynamic spatial organization of enzymes in the cell and the regulation of metabolism]. AB - The proteins in membranes and cytosol are often distributed unevenly in space and form the dynamic aggregates and functional assemblies. This property is important for functioning of multienzyme systems. The experimental results indicate the existence of a special type of regulation and integration of cellular metabolism based on the change of mutual rearrangement of enzymes in the cell and formation of transient protein assemblies. PMID- 3048424 TI - Changes in EEG mean frequency associated with anxiety and with amphetamine challenge in BPD. AB - Recent authors have hypothesized that cerebral dysfunction, as reflected in an abnormal EEG, may play an important role in the behavioral symptoms of patients with borderline personality disorder (BPD). Spectral analysis and amphetamine challenge testing are two promising methods for probing the clinical symptomatology of this disorder. In this study, we evaluated the relationship between clinical symptoms and computerized EEG spectral analysis in BPD patients both before and after amphetamine challenge. We found that mean frequency values on spectral analysis consistently correlated with anxiety levels in our patients, but did not correlate with a wide variety of other important symptoms, such as depression or transient psychosis. This result, coupled with our previous negative findings concerning EEG abnormalities in patients with BPD, casts doubt on the etiological relationship of cerebral dysrhythmias to the behavioral pathology of this disorder, but raises interesting questions concerning the relationship of anxiety and mean frequency. PMID- 3048425 TI - Obsessive-compulsive disorder: is there a frontal lobe dysfunction? AB - Obsessive-compulsive disorder has recently been found to be associated with various biochemical markers; this has revived interest in its biological basis. Most of the work to date has concentrated on the neurotransmitters involved. In this presentation, evidence from electrophysiological, neuropsychological, scan, lesion, and psychosurgical studies are integrated to focus on a possible frontal dysfunction in this disorder. PMID- 3048426 TI - Struwwelpeter Heinrich Hoffmann (1809-1894). PMID- 3048429 TI - National Institutes of Health consensus statement. Health implications of smokeless tobacco use. NIH Consensus Development Panel. AB - National data indicate that at least 10 million persons have used smokeless tobacco within the past year. The human evidence that use of snuff causes cancer of the mouth is strong. Smokeless tobacco use increases the frequency of localized gum recession and leukoplakia where the snuff is usually placed. The presence of lead in smokeless tobacco may pose a special risk for the developing fetus. Use of smokeless tobacco releases nicotine into the bloodstream and produces blood levels of nicotine comparable to those produced by smoking tobacco. The primary behavioral consequence of regular use of smokeless tobacco is long-term nicotine dependence and its associated health risks. PMID- 3048428 TI - Gliosis in schizophrenia. PMID- 3048427 TI - Hormone and metabolite plasma levels after oral glucose in bulimia and healthy controls. AB - Bulimia patients claim to crave sweets and since as clinical evidence suggests that the food consumed during eating binges often contains large amounts of carbohydrates, hormones involved in carbohydrate metabolism might be affected in bulimia. We therefore performed a 4-hr glucose tolerance test (GTT), using 100 g oral glucose and inquired about attitudes toward sweets. Thirteen female patients, with a mean age of 23.3 years, who had had bulimia from 3 to 7 years but whose binge-eating/vomiting behavior was largely controlled at the time of testing, were compared to 14 age-matched healthy female controls with a mean age of 24.4 years. All bulimic patients and most controls had liked sweets as children and still liked sweets. Significantly more bulimic patients than controls stated they overate on sweets and avoided sweets. Glucose utilization and the insulin, glucagon, growth hormone (GH), and pancreatic polypeptide (PP) response curves in the bulimic patients were within the normal range. Fasting plasma levels of glucose, insulin, glucagon, GH, cortisol, free fatty acids (FFA), and PP were not different from controls. There was a trend in bulimic patients to have lower plasma FFA levels and higher plasma cortisol levels during the GTT than controls. The findings suggest that, given body weight maintenance and adequate nutrition, patients with bulimia nervosa have normal glucose tolerance and normal hormonal responses following an oral glucose load. PMID- 3048431 TI - [Monoclonal antibodies ICO-20 against human thymocyte antigen T10]. AB - Monoclonal antibodies (Mab) Ig G2a isotypes reacting in indirect immunofluorescence assay with 68.7 +/- 4.1% of thymocytes, 7% of T-cells and not determining the antigen on other blood cells were obtained. Mab ICO-20 reacted in complement-dependent cytotoxic test. The antigen was expressed on colony-forming cells of granulocyte-macrophage row. Mab ICO-20 reaction with 100% of thymocytes was defined by flow cytometry. Antigen molecular mass is 45000 Dalton. The antigen was expressed on blast cells of patients with ALL and AML. Mab ICO-20 reaction was more more often with T-cell ALL. PMID- 3048430 TI - [Characteristics of endogenous proteolysis in preparations of human erythrocyte membranes]. AB - Endoproteolytic activity in human erythrocyte membrane preparations has been examined at 37 degrees C by one- and two-dimensional electrophoresis. Two dimensional mapping has shown that the presence of leukocyte enzymes in erythrocytes prepared in a regular manner (centrifugation) cannot be excluded. Sedimentation in the 1.5% dextran 500,000 with the following erythrocyte purification on HBS-cellulose has made it possible to prepare erythrocyte membranes characterized by low level endoproteolytic activity without leukocyte enzymes. The marker peptide has been found. It is likely to be a specific product of the enzyme activity of membrane localization. PMID- 3048433 TI - Molecular basis and prenatal diagnosis of beta-thalassemia. AB - The molecular characterization of mutations producing beta-thalassemia in world populations is nearing completion. We expect that new rare alleles in thoroughly studied groups and other alleles in less studied groups, eg, inhabitants of New Guinea, Latin America, and certain Pacific Islands, will be found. Knowledge of the molecular basis of the disease and new technology that allows rapid detection of single nucleotide changes in genomic DNA have led to the reality of prenatal diagnosis by direct mutation detection even in the heterogeneous US population. Programs aimed at prevention of beta-thalassemia should be facilitated by these developments. PMID- 3048434 TI - Enhanced survival but reduced engraftment in murine recipients of recombinant granulocyte/macrophage colony-stimulating factor following transplantation of T cell-depleted histoincompatible bone marrow. AB - In vivo administration of murine recombinant granulocyte/macrophage colony stimulating factor (rGM-CSF) was evaluated for effects on survival and engraftment in an allogeneic murine bone marrow transplantation (BMT) model involving T-cell depletion of donor marrow. The model provides a high incidence of graft failure/rejection. Recipients of continuous subcutaneous infusions of rGM-CSF had a significant survival advantage when compared with untreated controls. However, a significantly lower incidence of donor cell engraftment was noted. Hematological parameters were not substantially affected. When rGM-CSF was administered intraperitoneally (IP), twice daily injections closely approximated the effects of continuous infusion on survival. Single IP injections were without significant effects on survival or engraftment. These results demonstrate that prolonged frequent in vivo exposure to rGM-CSF can significantly improve survival but significantly decreases donor cell repopulation in recipients of T-cell depleted histoincompatible marrow grafts. PMID- 3048432 TI - [Chimeric drift in blood erythrocyte population in BALB/c----C57BL/10 and BALB--- B10.D2 mice]. AB - Genotypic composition of the erythrocyte population of peripheral blood in 16 aggregation chimeras: BALB/c (H-2dd)----C57BL/10(H-2bb) and BALB/c(H-2dd)--- B10.D2(H-2dd) was studied during 10 months. The proportion of cells of parental components was defined visually in the coat and by electrophoresis of allozyme variants at the Gpi-1 locus in blood. The similar increase of blood cells percentage was observed in blood of both types of chimeras with age. In chimeras the skin grafts of both parental types survive. Chimeric drift is not caused by H 2 haplotypes differences between cells of two strains or disturbance of immunological tolerance in the chimeric mice. We propose that chimeric drift results from interaction of hemopoietic cells of different strains in early stages of hemopoiesis. PMID- 3048436 TI - Lymphocyte depletion of donor bone marrow by counterflow centrifugal elutriation: results of a phase I clinical trial. AB - We report here the results of a phase I clinical trial using counterflow centrifugal elutriation (CCE) for the removal of donor T lymphocytes before allogeneic bone marrow transplantation (BMT). Thirty-eight patients received lymphocyte-depleted allografts from HLA-identical, MLR-nonreactive sibling donors. The patients entered onto the study were either at high risk on the basis of age (median, 39 years) or disease status (acute leukemia in early relapse [ER], chronic myelogenous leukemia [CML] in accelerated phase [AP], or therapy resistant [RES] lymphoma). All patients received a standard lymphocyte dose of 1 x 10(6) morphologic lymphocytes per kilogram ideal body weight (BW) and were maintained on cyclosporine A (CsA) for 170 days after BMT. Prompt engraftment occurred in 37 of 38 patients with a median time to absolute neutrophil count (ANC) greater than 500/microL of 18 days. Although acute graft-v-host disease (GVHD; clinical stage I or greater) was observed in 45%, it was limited to the skin in all but five patients. Survival was related to disease status at the time of BMT. Among patients with acute leukemia in first or second remission, CML in chronic phase (CP) or lymphoma in partial remission (PR), 64% are currently alive, in contrast to 31% of patients with acute leukemia in third remission or early relapse, CML in second CP or AP, or RES lymphoma. Median follow-up for all patients was 351 days (range, 105 to 711 days). We conclude that this procedure is safe and warrants further evaluation in a randomized efficacy trial. PMID- 3048435 TI - Identification of a second transforming gene, rasn, in a human multiple myeloma line with a rearranged c-myc allele. AB - Multiple myeloma is a disease characterized by a long, slowly progressive phase and a final, more aggressive one. Little is known about the mechanism of transformation of myeloma cells, although the clinical characteristics of the disease suggest a multi-step process. Recently, a myeloma cell line, NCI-H929, was isolated from a patient with aggressive preterminal disease and found to have a rearranged myc allele. This myeloma cell line has been further characterized in a focus formation assay to determine whether its unusual growth characteristics were associated with a second activated transforming gene. We now report that the NCI-H929 myeloma cell line has an activated rasn allele in addition to a rearranged myc allele. This is the first identification of an activated transforming gene in a multiple myeloma cell line; furthermore, the characterization of two independently activated oncogenes in this B cell malignancy has implications for both the pathogenesis and evolution of the disease. PMID- 3048437 TI - Granulocyte macrophage colony-stimulating activity production by cultured human thymic nonlymphoid cells is regulated by endogenous interleukin-1. AB - Supernatants of cultured human thymic nonlymphoid cells were assayed for granulopoietic factors using cultures of low density bone marrow mononuclear cells (LD-BMMC). Thymic nonlymphoid cell-conditioned medium (TNLC-CM) supported vigorous myeloid colony growth of LD-BMMC, and of LD-BMMC depleted of T lymphocytes and/or monocytes. Colony stimulating activity (CSA) in TNLC-CM was abrogated by a highly specific neutralizing antiserum against recombinant human granulocyte-macrophage colony-stimulating factor (GM-CSF). TNLC-CM also enhanced colony growth in LD-BMMC stimulated by colony stimulating activity from a giant cell tumor culture (GCT). The enhancing activity of TNLC-CM, unlike its CSA activity, required the presence of adherent cells in the marrow cell culture. The addition of anti-interleukin-1 (anti-IL-1) antibody to TNLC-CM inhibited the GCT enhancing activity, but not the CSA. When the anti-IL-1 immunoglobulin was added directly to cultures of thymic nonlymphoid cells, GM-CSF production was completely inhibited, and the GCT enhancing activity was neutralized. We conclude that an intercellular regulatory network exists in cultured thymic explants in which GM-CSF expression is induced by IL-1. In this system, the granulopoietic effect of IL-1 derives not from a direct effect on myeloid progenitors, but from its ability to recruit CSA production by other cells. PMID- 3048438 TI - A novel leukemia cell line, MR-87, with positive Philadelphia chromosome and negative breakpoint cluster region rearrangement coexpressing myeloid and early B cell markers. AB - We developed a Philadelphia chromosome (Ph) positive cell line, designated MR-87, from a 4-year-old boy with Ph+-acute leukemia. MR-87 cells grew in single cell suspensions with a doubling time of 120 to 144 hours. Both MR-87 and original leukemia cells were positive for myeloperoxidase (MPO) and myeloid antigen CD13. These cells exhibited the early B-cell phenotype, ie, terminal deoxynucleotidyl transferase+, Ia+, CD19+, and CD10+. Rearrangement of the immunoglobulin heavy chain was confirmed in both. Approximately 80% of MR-87 cells coexpressed CD13 and lymphoid antigens CD10 or CD19, as confirmed by a two-color analysis. Simultaneous expression of MPO and CD19 on a single MR-87 cell was demonstrated at ultrastructural level. Thus, MR-87 is a Ph+ leukemia cell line exhibiting a hybrid phenotype. The breakpoint cluster region (bcr) was not rearranged in the MR-87 cells and subsequent analysis using antisera revealed that these cells expressed a novel protein, P190c-abl, which was immunoprecipitated with anti-abl and anti-phosphotyrosine antibodies. The MR-87 line will be most useful for investigating the biology and pathogenesis of Ph+ bcr- acute leukemia. PMID- 3048439 TI - What radiation dose for DLA-identical canine marrow grafts? AB - In view of reported attempts at marrow grafting after nuclear accidents with a broad range of radiation exposures, the present study explored the total-body irradiation (TBI) conditions needed for engraftment in a canine model by using marrow from DLA-identical littermates. Previous studies have shown that such grafts are consistently successful when recipients are exposed to 920 cGy of TBI delivered at a rate of 7 cGy/min from opposing dual cobalt sources. The present TBI doses were all in the lethal range. Five dogs were administered 450 cGy; seven dogs, 600 cGy; five dogs, 700 cGy; and five dogs, 800 cGy of TBI administered at 7 cGy/min. They received a median of 3.3 x 10(8) marrow cells/kg intravenously after completion of radiation. Results showed transient allogeneic marrow engraftment in all dogs administered the lowest dose of TBI studied (450 cGy). Importantly, transient grafts permitted four of five dogs to live long enough for autologous marrow recovery to occur. At increasing radiation doses, 600, 700, and 800 cGy, the risk of graft failure lessened, with 3 of 7, 2 of 5, and 1 of 5 dogs, respectively, showing graft rejection. Fewer dogs survived with autologous marrow recovery, and more showed sustained allogeneic engraftment (4 of 7, 3 of 5, and 4 of 5 dogs, respectively). We conclude that DLA-identical littermate marrow grafts are beneficial in the setting of otherwise lethal radiation exposures, with most dogs either experiencing sustained allogeneic engraftment or surviving with autologous marrow recovery due to the extended support provided by a transient allogeneic graft. PMID- 3048440 TI - Neutrophil migration is defective during recombinant human granulocyte-macrophage colony-stimulating factor infusion after autologous bone marrow transplantation in humans. AB - We have previously reported that continuous intravenous (IV) administration of recombinant granulocyte-macrophage colony-stimulating factor (rHuGM-CSF) to humans following high-dose alkylating agent chemotherapy and autologous bone marrow support (ABMS) results in myeloid bone marrow maturation, accelerated granulocyte recovery, and reduced treatment-related toxicity. However, we found that leukocyte counts declined rapidly after discontinuation of rHuGM-CSF therapy, which suggests possible growth factor effects on leukocyte margination and migration. For these reasons we studied granulocyte margination by using 111In-labeled autologous granulocytes and found similar granulocyte margination before (21.5% +/- 13.4%) and during continuous IV rHuGM-CSF infusion (23.3% +/- 9.6%). Phagocytosis of Cryptococcus neoformans and granulocyte hydrogen peroxide production was similar before and during rHuGM-CSF infusion and similar to patients treated with the same high-dose chemotherapy and ABMS but not receiving growth factor. However, migration of granulocytes to a sterile inflammatory site was markedly reduced during continuous rHuGM-CSF infusion (1.2 +/- 0.9 WBCs/cm2, 24 hr) as compared with baseline (39.6 +/- 17.7 WBCs/cm2/24 hr; P less than .0008). These findings may be of relevance when extravascular granulocytes are required for host defense. PMID- 3048441 TI - Enhanced expression of the granulocyte-macrophage colony stimulating factor gene in acute myelocytic leukemia cells following in vitro blast cell enrichment. AB - Expression of the granulocyte-macrophage colony-stimulating factor (GM-CSF) gene in acute myelocytic leukemia (AML) was assayed by Northern blot analysis. GM-CSF messenger RNA (mRNA) was detected in the freshly obtained mononuclear cells of only one of 48 cases of AML, in contrast with recent reports that GM-CSF mRNA might be detected in half of the cases of AML when RNA is prepared from T-cell- and monocyte-depleted leukemic cells. We did find, however, that expression of the GM-CSF gene was detectable in five of ten cases after in vitro T-cell and monocyte depletion steps. Additional studies suggest that expression of GM-CSF in the bone marrow of the one positive case, rather than being autonomous, was under exogenous control, possibly by a paracrine factor secreted by marrow stromal cells. These studies emphasize the potential for altering in vivo patterns of gene expression by in vitro cell manipulation. PMID- 3048443 TI - Erythroid burst-promoting activity produced by interleukin-1-stimulated endothelial cells is granulocyte-macrophage colony-stimulating factor. AB - Interleukin-1 (IL-1) induces cultured human umbilical vein endothelial cells to elaborate heterogeneous hematopoietic growth factors, including granulocyte macrophage and granulocyte colony-stimulating factors (GM-CSF and G-CSF, respectively). Because erythroid burst-promoting activity (BPA) is also elaborated by endothelial cells exposed to IL-1, we sought to determine whether the BPA released by IL-1-induced endothelial cells simply reflects the known erythropoietic activity of GM-CSF or whether other uncharacterized factors might be involved. Media conditioned by multiply passaged endothelial cells cultured for three days with recombinant IL-1 alpha (ECMIL-1) stimulated erythroid burst and GM colony formation in cultures of human nonadherent T-lymphocyte-depleted marrow mononuclear cells. Pretreatment with an anti-GM-CSF antiserum neutralized all the BPA and 56% of the GM colony-stimulating activity (GM-CSA) in ECMIL-1. The antiserum used in these studies did not inhibit IL-3 or G-CSF activity and did not inhibit ECMIL-1-induced murine GM colony growth (a measure of human G CSF). To examine whether GM-CSF induces BPA release by accessory cells, media conditioned by marrow cells cultured for three days with GM-CSF were tested in the colony growth assays. Pretreatment with anti-GM-CSF antiserum completely neutralized the BPA and GM-CSA of the marrow cell-conditioned medium. We conclude that GM-CSF is the BPA elaborated by IL-1-induced endothelial cells. The in vitro erythropoietic activity of GM-CSF is not dependent on induced BPA release by accessory cells and therefore likely results from a direct effect of GM-CSF on progenitor cells. PMID- 3048442 TI - Pharmacokinetics of human granulocyte-macrophage colony-stimulating factor using a sensitive immunoassay. AB - A sensitive and reliable sandwich enzyme-linked immunosorbent assay (ELISA) has been developed for recombinant human granulocyte-macrophage colony-stimulating factor (hGM-CSF). The assay is quantitative between 100 pg/mL and 2.5 ng/mL for bacterially synthesized hGM-CSF in human serum and is more sensitive and specific than the semisolid agar bioassay. As part of a phase I study, the pharmacokinetics of intravenous (IV) bolus injection and subcutaneous (SC) administration of hGM-CSF were studied. Following a single IV dose, an initial high blood level of hGM-CSF occurred, followed by a rapid decrease occurring in two apparent phases with a half-life (t1/2)alpha of less than five minutes and a t1/2 beta of 150 minutes. After an SC injection, detectable serum levels occurred within 15 to 30 minutes, and serum levels were sustained for a variable time depending on the dose. At the highest SC dose (10 micrograms/kg), a serum level of greater than 1 ng/mL (65 pmol/L) was maintained for greater than 12 hours after a single injection. This corresponds to the concentration of hGM-CSF supporting near-maximum proliferation in vitro. PMID- 3048444 TI - Erythroid burst-promoting activity of purified recombinant human GM-CSF and interleukin-3: studies with anti-GM-CSF and anti-IL-3 sera and studies in serum free cultures. AB - We studied the erythroid burst-promoting activity (BPA) of recombinant human granulocyte/macrophage colony-stimulating factor (GM-CSF) and Interleukin-3 (IL 3) with two experimental approaches. First we studied the effects of polyclonal antisera prepared against human GM-CSF and gibbon IL-3 on colony formation from 1,000 bone marrow null cells/dish in serum-containing culture. Both GM-CSF and IL 3 independently enhanced erythroid burst formation; however, IL-3 showed more BPA activity than GM-CSF. These data are in agreement with an emerging view that the primary targets of IL-3 are primitive progenitors and that the targets of GM-CSF are intermediate progenitors, including erythroid burst-forming units (BFU-E). The proliferation of one population of BFU-E was independent of GM-CSF or IL-3. To characterize this population of BFU-E further, we developed a serum-free culture assay for the purified progenitors by incorporating insulin-like growth factor-1 (IGF-1) to the serum-free medium. The development of erythroid bursts was supported by IL-3, IGF-1, and erythropoietin (Ep) in a serum-free culture system and to a lesser extent by the combination of GM-CSF, IGF-1, and Ep. Although the burst-promoting ability of GM-CSF and IL-3 was again demonstrated in this system, unlike serum-containing culture Ep alone did not support burst formation. These results indicate that when fetal calf serum (FCS) is present, the culture system contains BPA that is not GM-CSF or IL-3. PMID- 3048445 TI - Effect of recombinant hematopoietic growth factors on proliferation of human marrow progenitor cells in serum-deprived liquid culture. AB - We investigated the effects of recombinant interleukin-3 (IL-3), granulocyte macrophage and granulocyte colony-stimulating factors (GM-CSF and G-CSF), and erythropoietin (Ep) on the number of human hematopoietic progenitors after two to ten days of incubation in liquid cultures deprived of fetal bovine serum (FBS). The source of progenitor cells was normal human marrow depleted of T lymphocytes and/or adherent cells. When adherent cell-depleted marrow was cultured without growth factors, the number of progenitor cells was relatively constant for periods up to eight days. In contrast, a progressive decline in the number of progenitor cells was detected in cultures of nonadherent, T-cell-depleted marrow cells. In both cases, the addition of IL-3 increased by two- to fourfold over input the number of erythroid burst-forming cells (BFU-E) per culture. The number of BFU-E peaked either at day 4 or 8. G-CSF had no effect on the number of progenitor cells per culture. GM-CSF and Ep had no effect in cultures of nonadherent marrow cells but maintained the number of BFU-E in cultures of nonadherent, T-cell-depleted marrow cells. The addition of a neutralizing anti-GM CSF monoclonal antibody, but not anti-IL-3 neutralizing antiserum, decreased the number of BFU-E in cultures of nonadherent marrow cells. None of the growth factors investigated enhanced the number of GM progenitors to the same degree as the number of BFU-E. However, in cultures of nonadherent, T-cell-depleted marrow cells, IL-3 and GM-CSF maintained the number of GM progenitors up to eight days. These results indicate that IL-3 alone is capable of increasing the number of BFU E and of maintaining the number of GM progenitors in liquid culture, whereas GM CSF and Ep are capable of maintaining, but not increasing, BFU-E in this system. PMID- 3048446 TI - Differential expression of LFA-1 molecules in non-Hodgkin's lymphoma and lymphoid leukemia. AB - We investigated the expression of adherence molecules lymphocyte function associated antigens-1 alpha and -beta (LFA-1 alpha, -beta) and p150, 95 in 103 well-characterized non-Hodgkin's lymphomas (NHLs) and lymphoid leukemias (LLs). We found that NHLs and LLs differentially express LFA-1 molecules according to their lineage derivation, degree of clinical aggressiveness, and anatomic site of involvement. Specifically, (a) T-cell neoplasms nearly always express these molecules; (b) diffuse aggressive B-cell NHLs and mature LLs often lack LFA-1 alpha molecules; and (c) B-cell chronic lymphocytic leukemia (CLL) is often LFA-1 alpha-negative while B-cell small lymphocytic lymphomas (SLLs) are nearly always LFA-1 alpha-positive. Furthermore, the low expression of LFA-1 alpha in CLL is related to the low degree of homotypic lymphocyte adhesion after tumor promoter antigen stimulation that does not modulate the expression of LFA-1 alpha in vitro. The differential expression of LFA-1 by B-cell CLL and SLL and their degree of homotypic lymphocyte adhesion may account for the distinct anatomic compartmentalization and characteristic clinical behavior of these two morphologically and immunologically similar lymphoid malignancies. PMID- 3048448 TI - Techniques related to the clinical epidemiology of peptic ulcer disease. PMID- 3048449 TI - Physiology and pharmacology of the parietal cell. AB - The parietal cells possess the unique capacity to produce large quantities of acid at a high concentration, and this is reflected in unique properties at the cellular level. The cells are comparatively large, and they are equipped with secretory canaliculi, a multitude of mitochondria, and cytoplasmic tubulovesicles. During secretion many of the tubulovesicles merge with the secretory canaliculi, which then expand. In the process H+, K+-ATPase is transferred from the tubulovesicular membrane to the secretory membrane. This enzyme catalyses the final step in the production of HCl. Parietal cell activity is regulated through receptors on the basolateral cell surfaces. In the isolated gland and in the isolated parietal-cell fractions, stimulation of receptors for histamine evokes higher secretion than receptor stimulation with cholinergic compounds or with gastrin. In these experimental models, specific inhibitors are required to block acid secretion; for example histamine H2-receptor antagonists will block histamine-induced secretion but will be inactive when secretion is evoked by gastrin or by cholinergic stimulation. These stimuli cause a more or less marked increase in the intracellular levels of Ca2+, which acts as a second messenger, leading to the activation of phosphokinases and, ultimately, to morphological transformation of the parietal cells and acid secretion. Another such intracellular messenger is cAMP, which is formed in response to histamine stimulation only; prostaglandins may prevent this process and block acid secretion. The final step in the production of acid requires K+ and Cl- channels in the secretory membrane and the H+, K+-ATPase-catalysed exchange of K+ for H+ across this membrane. This reaction consumes large amounts of energy and depends on the aerobic production of ATP by the parietal cells. Substituted benzimidazoles, such as omeprazole, accumulate in the acid compartments of the parietal cells and inhibit the H+, K+-ATPase, thereby blocking acid production. PMID- 3048450 TI - Aetiology of ulcers. AB - It seems that duodenal and gastric ulcers are caused by environmental ulcerogens, which are probably infectious or chemical. The reasons for individual susceptibility to these ulcerogens have not been defined and, indeed, it is not yet certain that the effects are not essentially random. Abnormalities of function of the mucosae of the upper alimentary tract do not appear to be necessary or sufficient for the production of ulcers. The two principal clinical aspects of ulcer disease--the tendency to form chronic mucosal wounds and the tendency of the wounds to recur during many years--point to, but cannot yet be explained in terms of, failure of the processes involved in wound repair. More specifically, it is not known whether there is interference with the processes involved in normal mucosal repair or whether there is failure of the repair processes. When these problems are closer to solution, it will perhaps be possible to assess how environmental factors influence ulcerogenesis. PMID- 3048451 TI - Bacteria in ulcer pathogenesis. AB - A great deal of information about the spiral bacteria of the stomach has accumulated in the past 5 years. These bacteria, currently named Campylobacter pylori but likely to be renamed as a new genus, have adapted to living beneath the mucus layer and above the gastric surface mucous cells. When metaplastic gastric mucous cells are also present in the duodenal bulb, C. pylori may also get a foothold in this latter location. Observations of the high prevalence of C. pylori in patients with gastritis and with duodenal ulcers, and the slightly lower prevalence in patients with gastric ulcer, have led to the hypothesis that the bacteria play an aetiological role in these three conditions. There is now fairly convincing evidence that the organisms can cause active chronic gastritis. The most persuasive of this comes from reports of the rapid development of gastritis and symptoms in two volunteers who swallowed the organism, plus two other series of accidental challenges. Other evidence is provided by the waning and waxing of gastritis, which has been correlated in several studies with clearance followed by recrudescence of the organisms. The role of the bacterium in peptic ulcer is less certain. The present data do not provide strong evidence for a causal role in gastric ulcer, although we cannot rule out that it may be important in some. The very high prevalence in patients with duodenal ulcer, including one series in children (who rarely harbour the organism), raises the distinct possibility that the bacteria play an aetiological role in this form of ulcer. Reports of ulcer healing with antibiotics and of lower recurrence rates in those cleared of the organism, increase the possibility, However, methodological flaws in some studies, plus the usual need for confirmation of key studies, indicate that we should await more definitive evidence before accepting that duodenal ulcer can be an infectious disease. PMID- 3048452 TI - Site-protective agents. PMID- 3048447 TI - The development of tamoxifen for breast cancer therapy: a tribute to the late Arthur L. Walpole. AB - Tamoxifen, a nonsteroidal antiestrogen, is now the endocrine treatment most widely used in breast cancer, both in the adjuvant and advanced disease settings. Here we will trace the development of tamoxifen for advanced breast cancer in postmenopausal patients, consider the biological basis for the recent successful use of tamoxifen for long-term adjuvant therapy, and discuss the use of tamoxifen in premenopausal patients with advanced disease. In part, this will be a historical review offered as a tribute to the late Dr. Arthur L. Walpole, who must receive the chief credit for the discovery of tamoxifen and its subsequent application as an anticancer agent. PMID- 3048454 TI - Omeprazole. PMID- 3048455 TI - Antacids: the past, the present, and the future. AB - Antacids have served us well for over a century. The attitude in the late 1950s to 1970s that antacids should be taken only on demand was unjustified and was based on what can be seen nowadays as misinterpretation of scientific data. Twelve recent endoscopic controlled studies have confirmed the efficacy of antacids in the healing of duodenal ulcer, achieving about 75% healing in 4 weeks. Like H2-receptor antagonists, the efficacy of antacids in the healing of gastric ulcers is controversial, most probably related to the even greater pathogenetic heterogeneity of this condition. Antacids should be given at least four times a day and at least 1 hour after meals, since their therapeutic success most likely depends on neutralization of postprandial acid secretion. In vivo, the newer tablet forms are indistinguishable from the liquid forms in terms of neutralizing efficiency and healing efficacy. The ideal dose is one that neutralizes 400 mmol of acid. Combination with an anticholinergic drug is effective and a recent report suggests that this may lead to longer remission than with H2-receptor antagonists. As a long-term therapy, antacids appear to work but need to be taken in multiple daily doses, a regimen which is unlikely to meet with long-term patient compliance. The success of antacids in duodenal ulcer healing should alert us to the importance of controlling the meal-stimulated acid secretion in ulcer therapy, and to the hard fact that acid is a non-permissive factor in ulcer healing. PMID- 3048453 TI - Prostaglandin analogues. PMID- 3048456 TI - Medical regimens in short- and long-term ulcer management. PMID- 3048458 TI - Surgery and its sequelae. PMID- 3048457 TI - Diagnosis and treatment of Zollinger-Ellison syndrome. AB - A diagnostic and therapeutic strategy for the management of patients with Zollinger-Ellison syndrome has been developed, based on the review of a large personal experience and the most recent literature. The mainstay of a modern ZES management is the eradication of tumoral processes whenever feasible. Diagnosis is centred upon gastric acid and gastrin secretion measurements both in basal conditions and on secretin stimulation. Recognition of other endocrine involvement and familial inheritance is of the utmost importance in distinguishing sporadic ZES patients from those who have the condition known as multiple endocrine neoplasia type I. Blood calcium and phosphorus levels, parathyroid hormone concentration, combined if necessary with urinary cyclic AMP excretion measurement, should be performed routinely once ZES diagnosis is established or highly suspected. Localization of the tumour is the next essential step, and this has been considerably facilitated by the recent development in imaging techniques: it involves computerized axial tomography and selective abdominal angiography, a combination of which allows tumour detection in 60-70% of sporadic gastrinoma patients, with a maximal sensitivity for well-developed hepatic metastases. In sporadic ZES exploratory laparotomy is legitimate when preoperative localization of the tumour has failed; this laparotomy will allow further detection and then eradication of gastrinomas in a significant number of patients. Control of gastric acid secretion is mandatory throughout the work-up period; modern antisecretory agents are efficacious in most cases; total gastrectomy, when control of acid hypersecretion has failed, is now exceptional. Eradication of the tumour should be attempted in cases of sporadic ZES in the absence of recognizable liver involvement. The chance of a definite cure provided by surgery when performed by an experienced surgeon varies from 20% to 60% in pancreatic and ectopic gastrinomas respectively. In ZES patients with MEN I, exploratory laparotomy is seldom indicated (other than for symptomatic associated endocrine secretion), as the chance of a definite cure by surgery is very rare. Parathyroid surgery is often indicated and should take place before any form of abdominal surgery. In cases of hepatic metastases, chemotherapy with streptozocin and fluorouracil is indicated and soon, perhaps, chemo-embolization. PMID- 3048459 TI - Entrepreneurs and private enterprise: the development of medical lecturing in London, 1775-1820. PMID- 3048461 TI - Obsessive questions and faint answers: the French response to tuberculosis in the Belle Epoque. PMID- 3048460 TI - Alexander Monro primus and the Edinburgh manner of anatomy. PMID- 3048462 TI - Federal intervention in the Joplin silicosis epidemic, 1911-1916. PMID- 3048463 TI - Mom and Dad (1944): venereal disease "exploitation". PMID- 3048465 TI - How can results of bone marrow transplantation be improved? PMID- 3048464 TI - Drug-induced ototoxicity. PMID- 3048466 TI - Second marrow transplants in patients with leukemia who relapse after allogeneic marrow transplantation. AB - Twenty-six patients with recurrent leukemia following allogeneic marrow transplantation received a second marrow transplant between 1.5 and 78 months (median 26) after the initial transplant. Preparative regimens for second transplant included multi-agent chemotherapy with total body irradiation, 2.0 10.0 Gy (five patients), dimethylbusulfan alone (one patient), and dimethylbusulfan or busulfan plus cyclophosphamide (20 patients). One patient died before engraftment of infection and 18 died after engraftment from veno occlusive disease (4), infection (2), idiopathic pneumonia (3), cytomegalovirus pneumonia (3), leukemia (5) and encephalopathy (1). Seven patients (27%) survive 12-38 months (median 26); five (19%) are disease-free and two have recurrent leukemia. Two of the five disease-free survivors have chronic graft-versus-host disease. All of the surviving patients received dimethylbusulfan or busulfan plus cyclophosphamide and six of the seven surviving patients were among 11 patients transplanted more than 2 years after the first transplant whereas only one was among the 15 transplanted in less than 2 years. Those who have second marrow transplants one or more years after their initial transplant are more likely to benefit, while those who are less than 1 year from initial transplant appear to benefit the least. PMID- 3048469 TI - Oral condition in children treated with bone marrow transplantation. AB - Oral health was studied in 49 children below 12 years of age treated with bone marrow transplantation (BMT). Dental treatment prior to BMT was required in 41% of the children. Ulceration of the mucous membranes in the mouth was observed in 37% of the patients during treatment. Those given methotrexate as prophylaxis against graft-versus-host disease (GVHD) exhibited oral ulcerations more frequently (p less than 0.05) than those given cyclosporin. One year after BMT all patients with a clinical chronic GVHD exhibited changes in the oral mucosa. During the first year after BMT 31% of the children developed carious lesions. Children treated with 10 Gy total body irradiation (TBI) had a significantly (p less than 0.05) reduced salivary secretion rate. Dental development was severely affected in children treated with TBI. These disturbances consisted of arrested root development, enamel hypoplasias and microdontia. PMID- 3048468 TI - Immunomagnetic removal of B-lymphoma cells from human bone marrow: a procedure for clinical use. AB - B-lymphoma cells were purged from human bone marrow by incubating the cell suspension with a cocktail of three different pan-B cell mouse IgG1 monoclonal antibodies, and then with immunobeads charged with sheep anti-mouse antibody, followed by magnetic separation. The primary antibodies used, HD37 (CD19), HD6 (CD22), and HH1 (CD37), bind to a very high percentage of the cells in non Hodgkin's lymphomas of poor prognosis. The secondary antibody is directed against the Fc portion of the IgG antibodies. In model experiments Burkitt's lymphoma cells (Rael) were admixed to mononuclear bone marrow cells in the ratio 1/9. With a ratio of immunobeads/total antibody-binding B cells of 50/1 in a first treatment cycle and repeating the procedure with the same number of beads in a second cycle, a tumor cell depletion of more than 5 logs was achieved, as judged by a clonogenic assay. The concomitant reduction of CFU-GM and CFU-GEMM was about 20%. The purging procedure has been scaled up to clinical use. Equipment suitable for purging patients' marrow specimens, employing standard transfusion facilities, is described. With this equipment the efficacy of tumor cell removal was the same as in the model experiments, and the whole magnetic separation could be completed in 2 hours. PMID- 3048467 TI - Melphalan and total body irradiation (TBI) versus cyclophosphamide and TBI as conditioning for allogeneic matched sibling bone marrow transplants for acute myeloblastic leukaemia in first remission. AB - Between June 1981 and April 1986 63 patients with acute myeloid leukaemia (AML) in first remission and HLA-identical sibling donors were entered into a prospective study comparing cyclophosphamide (CY) + total body irradiation (TBI) with melphalan + TBI as conditioning therapy prior to transplantation. Thirty-six patients received CY/TBI and 27 received melphalan/TBI. The actuarial probability of remaining in remission for patients receiving melphalan/TBI was 94% compared with 66% following CY/TBI (p greater than 0.01). By including a comparable group of 41 patients with AML in first remission, conditioned with CY/TBI prior to the onset of the study, the greater anti-leukaemic effect of melphalan/TBI compared to CY/TBI was unchanged and statistically the chance of this being wrong is 1 in 10 (p less than 0.1). The overall survival in remission of both arms of the study was the same with 15/27 patients (55%) surviving in remission following melphalan/TBI compared with 19/36 patients (53%) following CY/TBI. The benefit obtained in reduced relapse was offset by the combined nephrotoxic effect of melphalan and cyclosporin which was not identified until the programme had been underway for a period of time. This shows that misinterpretation of 'no survival advantage' for the new treatment may occur due to unforeseen and preventable toxicities. PMID- 3048470 TI - The clinical diagnosis of acute graft-versus-host disease: a diversity of views amongst marrow transplant centers. AB - The diagnosis and grading of acute graft-versus-host disease (AGVHD) is based on a spectrum of clinical and laboratory features. To evaluate the reliability of current diagnostic and scoring criteria, six clinical vignettes were evaluated by 49 transplant physicians from 42 bone marrow transplant center worldwide. Responses were analysed to determine: (1) the degree of concordance among transplanters in diagnosing, scoring and treating AGVHD, and (2) the degree of concordance in assigning primary and contributing causes of death in patients with multiple complications of bone marrow transplantation. Concordance for the diagnosis of AGVHD ranged from 24 to 76%; concordance for grading AGVHD was 55%. Concordance for the decision to treat for AGVHD ranged from 43 to 55%; concordance for assigning the primary cause of death ranged from 71 to 100%. The findings demonstrate the need for (1) improved uniformity of reporting complications of bone marrow transplantation; (2) periodic revision and validation of diagnostic and grading criteria, and (3) methods of analysis that allow for multiple causes of death. PMID- 3048471 TI - Veno-occlusive disease of the liver following high-dose chemotherapy and autologous bone marrow transplantation in children with solid tumors: incidence, clinical course and outcome. AB - Two hundred and thirty-six consecutive courses of high-dose chemotherapy with autologous bone marrow transplantation in children with solid tumors were reviewed in order to assess the incidence, clinical presentation and outcome of veno-occlusive disease (VOD) of the liver. Patients conditioned with total body irradiation were excluded from this study. Eleven patients (4.6%) met the diagnostic criteria for VOD. The clinical course included sudden weight gain, jaundice, hepatomegaly and ascites. Renal dysfunction and refractoriness to platelet transfusions occurred in the most severe forms. Seven patients recovered within 7-29 days of onset and four patients died, all with renal failure and fluid overload. The time of onset appeared to determine two patterns of outcome: mild forms with early onset (before day 11) and more severe forms with onset after day 17. Analysis of pretransplant factors revealed no significant association with an increased risk of VOD. However, all the patients with severe VOD had received a conditioning regimen containing cyclophosphamide which might be involved in the pathogenesis of VOD. PMID- 3048472 TI - Post bone marrow transplant sera and persisting mast cell colonies. AB - Serum samples were collected from eight recipients of allogeneic bone marrow transplants (BMT) following the normalization of peripheral blood counts. Three patients (11 samples) were in the early engraftment period (less than 6 months post-BMT) and still receiving cyclosporin A or methotrexate and the remaining five patients (18 samples) were in stable engraftment (12-84 months post-BMT) and not on immunosuppressive therapy. All post-BMT serum samples supported the growth of increased numbers of mast cell colonies in long-term agar cultures when compared with normal controls (p less than 0.025-0.005), irrespective of the time from BMT or the presence of clinical graft-versus-host disease (GVHD). In contrast, the numbers of neutrophil, monocyte and eosinophil colonies were equivalent in test and control groups. It is proposed that post-BMT sera contain higher levels of a mast cell stimulating activity(s) than does normal sera. Such increased levels might be associated with clinical or subclinical GVHD. PMID- 3048473 TI - Psychological aspects of bone marrow transplantation: a retrospective study of 17 long-term survivors. AB - In a retrospective study 17 patients were interviewed 1-5 years after bone marrow transplantation (BMT) about their emotional experiences and about the information and support given to them. In all patients the illness, the BMT and its consequences created severe emotional strain; their psychological state was closely linked to their physical condition. Yet, most patients showed a surprising emotional elasticity and affective adjustment. Although the patients were overwhelmingly positive about the information and support they had received, an important number of them felt inadequately prepared for the emotional and sexual problems they had to face in the first period after discharge. The results of this study have led to structural changes in the psychiatric and psychosocial approach to the BMT patients. PMID- 3048474 TI - Repeated courses of high dose melphalan and unpurged autologous bone marrow transplantation in children with acute non-lymphoblastic leukemia in first complete remission. AB - Eleven children between the ages of 1 and 16 years with acute non-lymphoblastic leukemia (ANLL) in first remission were included in a study of double unpurged autologous bone marrow transplantation (ABMT). Prior to each ABMT patients received massive chemotherapy with melphalan at a dosage of 140 mg/m2. The first ABMT was done within a median of 4 months after the achievement of complete remission. As soon as the children had adequate hematologic recovery, a second marrow collection was done, followed by a second course of melphalan and a second ABMT. The duration of aplasia was significantly longer after the second ABMT than after the first, but the non-hematologic toxicity was relatively mild in each case and no patient died from the procedure. Four patients relapsed and seven are alive in unmaintained complete remission with a median duration of leukemia-free survival of 29 months (range 15-56 months) after the first ABMT. These data demonstrate the feasibility of repeating ABMT after melphalan in children with ANLL. The eventual impact of such therapy needs to be demonstrated in prospective randomized studies. PMID- 3048475 TI - Prognostic significance of cytogenetic abnormalities in patients with chronic myelogenous leukemia. AB - The cytogenetic data for 126 patients with Ph-positive chronic myelogenous leukemia (CML) in accelerated phase or blast crisis were analysed for clonal chromosomal abnormalities in addition to the standard Ph prior to allogeneic or syngeneic bone marrow transplantation (BMT). Additional clonal abnormalities were found in 84%, and 14% had a variant Ph (VPh). In decreasing order of frequency, the most common clonal abnormalities were a second Ph, +8, i(17q), -Y and +19. A second Ph, VPh or +8 occurred more frequently in patients who relapsed following BMT than in those who survived disease-free for at least 1 1/2 years. The presence of an i(17q) alone did not correlate with relapse. The patients with a second Ph, VPh or +8 had a median time to relapse of 19 months, and the risk of relapse at 3 years was 73%. Those with other or no additional clonal abnormalities had not reached a median time to relapse and had a 3-year risk of relapse of 31% (p = 0.002). This analysis suggests that specific cytogenetic abnormalities may be useful indicators of resistance to therapy for CML and should be included in proportional hazard models to predict outcome after BMT. PMID- 3048476 TI - Administration of human urinary colony stimulating factor after bone marrow transplantation. AB - A phase II study of the use of colony stimulating factor derived from human urine (CSF-HU) was performed after bone marrow transplantation (BMT). Steady and rapid recovery of leukocyte and granulocyte numbers was observed. In most patients who received CSF-HU from day 1, leukocyte numbers started to increase on day 6 and monocytes and granulocytes on day 11. Significant differences in the days to recovery of leukocytes over 1 x 10(9)/l and of granulocytes over 0.5 x 10(9)/l were observed in comparison with non-randomized control patients. In some patients in whom recovery of leukocytes was delayed, CSF also seemed to be effective in increasing leukocyte numbers from 8 days after the start of administration. There was no significant difference in the rate of relapse of leukemia between the two groups. CSF-HU seems very promising as a treatment of patients after BMT by shortening the period of leukopenia or granulocytopenia. PMID- 3048477 TI - Preliminary characterization of functional residual host-type T lymphocytes following conditioning for allogeneic HLA-matched bone marrow transplantation (BMT). AB - Host T lymphocytes which escape the effects of chemoradiotherapy may proliferate and lead to graft rejection, particularly in recipients of T cell-depleted bone marrow (BM) allografts. We studied the efficacy of several conditioning regimens including a new immunosuppressive regimen--total lymphoid irradiation (TLI) plus conventional chemotherapy and total body irradiation (TBI)--in abrogating residual host T lymphocytes as assessed by their ability to grow in vitro. A total of 38 patients were evaluated, 29 with hematologic malignancies, six with severe aplastic anemia (AA) and three with beta-thalassemia major (TM), of whom 32 were transplanted with HLA-identical T cell-depleted allogeneic BM from sibling donors. The median observation period was 15 months (range 3-21) posttransplant. Peripheral blood mononuclear cells (PBMC) taken from each patient on the day of transplant were cultured with interleukin 2 (IL-2) + phytohemagglutinin + irradiated donors' PBMC. Survival of cells for less than 2 weeks in vitro without proliferation was observed in 20 of 29 cases with malignancies and was considered negative. In this group only two leukemia (L) patients rejected the graft. Limited cell growth (less than or equal to 3 weeks) was seen in four L patients, two of whom showed early graft failure. Vigorous T cell growth (greater than 5 weeks, 62-96% CD4+ cells) was noted in eight cases (two L, four AA, two TM; none received TBI). In this group, sustained engraftment was observed in 7/7 patients who were treated with cyclosporin A (CSA) post grafting. Overall, we could demonstrate no clear correlation between graft failure and cell growth in vitro. The proliferating cells exhibited considerable cytotoxic activity in vitro against several tumor cell lines and were susceptible to pharmacological doses of CSA. The low incidence of continuous T cell proliferation in vitro in PBMC of L patients suggests that a combination of TLI, TBI and cyclophosphamide (CY) is highly effective in abrogating the host T cells and subsequent graft rejection. Since a rather small number of patients was included in this study, further studies are needed to determine the possible value of the in vitro T cell proliferation assay as a means for predicting graft failure. PMID- 3048478 TI - In utero bone marrow transplantation of fetal baboons with mismatched adult marrow: initial observations. AB - Recent advances in prenatal diagnoses of sickle cell anemia and thalassemia permit early identification of affected fetuses. However, the only intervention possible to date is abortion of the affected fetuses. Transplantation of normal marrow into fetuses in utero could correct these life-threatening disorders, but to accomplish this techniques must be developed for fetal transplantation in man. Therefore, we have transplanted fetal baboons with mismatched adult baboon bone marrow from donors that differed at the glucose phosphate isomerase locus. Twenty two fetuses between 60 and 160 days of gestation (term gestation is 182 days) were transplanted intraperitoneally with 10(9) marrow mononuclear cells/kg body weight using an ultrasonic technique. No immunosuppressive or preparative regimen was given prior to or after transplantation, and all fetuses tolerated the procedure well. One month after transplantation fetal blood samples were obtained to assess chimerism. Three chimeras were detected among 10 fetuses transplanted at 80 days' gestation, and no chimeras were detected in fetuses greater than 80 days' gestation at the time of transplantation. All chimeras died in utero during the third trimester of pregnancy: one of an intrauterine infection at 160 days' gestation, one at 135 days' gestation and one at 145 days' gestation. In contrast, the other 19 non-chimeric fetuses survived. These data suggest: (1) in utero fetal bone marrow transplantation is technically feasible in primates; (2) that allogeneic adult bone marrow can engraft and persist for at least 1 month in fetal baboons transplanted at 80 days of gestation; and (3) that delineation of the causes for loss of fetal chimeras should prove valuable in assessing the therapeutic potential for in utero bone marrow transplantation in man. PMID- 3048479 TI - Frequency analysis of cytotoxic T lymphocyte precursors--possible relevance to HLA-matched unrelated donor bone marrow transplantation. AB - HLA-matched unrelated donor (MUD) bone marrow transplants and transplants between HLA mismatched family members are associated with an increased incidence and severity of graft-versus-host disease (GVHD) in comparison with HLA-identical sibling transplants. A limiting dilution analysis system was set up to measure the frequency of alloreactive cytotoxic T lymphocyte precursors (CTL-p) in normal individuals and in potential donor/patient pairs selected for bone marrow transplantation. The donor/recipient pairs were divided into four groups depending on their degree of HLA disparity. A distinct range of CTL-p frequencies was obtained for each group and these showed a hierarchy of response related to the degree of HLA disparity between donors and recipients in that particular group. This assay system may be of value in selecting potential matched unrelated and mismatched family donor/patient pairs for those at lower risk of GVHD. PMID- 3048480 TI - Transient engraftment of T cell-depleted allogeneic bone marrow in mice improves survival rate following lethal irradiation. AB - C3H/HeJ mice were exposed to 8 or 9 Gy total body irradiation prior to transplantation of 1-15 x 10(6) H-2 incompatible T cell-depleted bone marrow cells from C57BL/6 donors. Survival was greatly enhanced compared to irradiated controls, even at the lowest cell doses. Analysis of spleen cells 1-7 weeks post transplant revealed that recipients of the lowest doses of T cell-depleted bone marrow had only transient engraftment of donor type cells, and that long-term recovery was autologous. This transient engraftment had a marked beneficial effect both on reconstitution of hematopoiesis and on survival. PMID- 3048481 TI - Q fever following bone marrow transplantation. AB - We describe a patient who developed an acute febrile illness 85 days after receiving an allogeneic bone marrow transplant for acute myeloid leukaemia. Serological investigation indicated the cause to be Q fever. The patient was receiving concurrent immunosuppressive therapy for graft-versus-host disease (GVHD). Coxiella burnetii should be added to the list of organisms which may complicate bone marrow transplantation. This case also provides further evidence to support an association between immunocompromised conditions and the development of Q fever. PMID- 3048483 TI - Blood cell changes after radiation exposure as an indicator for hemopoietic stem cell function. AB - This paper describes the criteria to be used in the management of persons accidentally exposed to ionizing radiation for predicting whether the stem cell pool damage was reversible or irreversible. This question is of importance. If the damage was reversible, the clinical management may be restricted to symptomatic therapeutic measures (antibiotics, platelet transfusions). If the indicators show that the stem cell damage is irreversible (from a clinical viewpoint) then stem cell transplantation must be considered and performed. A granulocyte computer simulation model is discussed that may be useful in the analysis and evaluation of blood cell regeneration patterns after radiation and transfusion of stem cells from different sources. PMID- 3048482 TI - A case of accidental cyclosporin overdose with pharmacokinetic analysis. AB - In a case of oral overdose of cyclosporin we measured plasma concentrations by radioimmunoassay and high pressure liquid chromatography. We observed unchanged pharmacokinetic parameters after the overdose indicating normal renal and hepatic function. Laboratory findings showed no renal or hepatic damage. As toxic symptoms, only emesis and a short, mild drowsiness were noted. PMID- 3048484 TI - Marrow transplantation in the treatment of murine hereditary diseases. AB - The prospects for analysing the curative potential of mutant stem cells following transfection with wild type genes is severely limited by the number of available animal models that fulfil all the requirements. Appropriate models must have the following characteristics: (1) the wild type stem cells are capable of providing a long-term cure of the mutant; (2) the primary product of the mutated gene is known; (3) the wild type allele is cloned; and (4) the normal gene is capable of being reinserted into mutant bone marrow stem cells where it continues to function when the cells are returned to the mutant environment. My object in this paper is to review stem cell transplantation in Mus musculus with hereditary anomalies, thus identifying the mutant mice that fulfil the first criterion and are potential models for future gene transfection studies. PMID- 3048485 TI - T cell depletion in bone marrow transplantation for leukemia: current results and future directions. AB - T cell depletion reduces the incidence and severity of graft-versus-host disease (GVHD) following bone marrow transplantation in man. Graft-versus-host disease of more than grade 2 severity is decreased from about 45% to about 10% in recipients of HLA-identical transplants. However, T cell depletion also increases the frequency of graft failure and leukemia relapse. Graft failure increases from about 1 to 10% following HLA-identical transplants. In patients with acute leukemia in first remission or with chronic myelogenous leukemia in chronic phase, leukemia relapse increases from about 20% to about 40%. Thus, although T cell depletion decreases GVHD, it increases graft failure and leukemia relapse such that survival is not convincingly improved. Several approaches to these problems are possible including increased pre- or post-transplant immune suppression, more effective antileukemia therapy or selective T cell depletion. Preliminary results of these approaches are discussed and new directions suggested. PMID- 3048486 TI - Cytokines as mediators of graft-versus-host disease. AB - Cytokines are proteins produced mainly by lymphocytes in response to an antigenic stimulus. Originally identified and named on the basis of their biological activity, they are now called interleukins; together with the interferons, colony stimulating factors and tumour necrosis factor/cachectin (TNF) they form a complex and overlapping network of communication between immunocompetent cells. Cytokines play a central role in T cell activation, and interleukin 2 and interferon gamma in particular are involved in the expression of graft-versus host disease after bone marrow transplantation. Recent studies suggest that TNF is also implicated: the gene encoding TNF is situated close to the MHC gene in both mice and humans, and TNF is able to up-regulate constitutively expressed class II antigen and, with interferon gamma, to induce class II expression in previously normal cells. Bacterial lipopolysaccharide (endotoxin) is a powerful stimulus to TNF, and TNF production may be the mechanism underlying the longstanding observations on the role of the bacterial microflora of the gut in graft-versus-host disease. PMID- 3048487 TI - Optimal time interval between myeloablative whole body irradiation and reconstitution with syngeneic bone marrow graft. AB - The optimal time interval between termination of radiation therapy and marrow grafting has been studied in an animal model of syngeneic bone marrow transplantation, using a limiting number of bone marrow cells. Optimal survival was achieved when reconstitution took place 24 h following termination of radiation therapy at all cell doses studied, including 5 x 10(4), 2 x 10(5) and 1 x 10(6) cells/recipient. The worst results were observed when bone marrow cells were infused immediately after irradiation. These results suggest that reconstitution of lethally irradiated recipients requires migration of irradiated host marrow cells and establishment of adequate 'bone marrow space', which is optimal at 24 h following termination of whole body irradiation. PMID- 3048488 TI - Alternative donor sources in HLA-mismatched marrow transplantation: T cell depletion of surgically resected cadaveric marrow. AB - Allogeneic bone marrow transplantation has been largely confined in its application to patients with an HLA-matched marrow donor, thereby limiting potential therapeutic benefits for patients with diseases amenable to treatment by marrow transplantation who lack effective alternative therapies and an identified matched marrow donor. T cells in the donor marrow generate graft versus-host disease which is a major consideration in attempts to widen the application of allogeneic marrow transplantation to the minimally HLA-matched or mismatched setting. The method of marrow harvest influences both the number of such T cells in the donor marrow inoculum and the functional capacity of T cells in murine marrow. We have therefore evaluated surgically resected cadaveric marrow for T cell content and for T cell depletion. Surgically resected marrow is more easily depleted of T cells than aspirated marrow and clinically useful quantities of marrow have been obtained and cryopreserved in a bank of characterized donor marrow for future use in HLA minimally matched or unmatched marrow transplantation. PMID- 3048489 TI - Aspergillus coronary embolization causing acute myocardial infarction. AB - An increased frequency of disseminated aspergillosis has been observed in the last decade, mostly occurring in immunocompromised patients including the bone marrow transplant population. Cardiac involvement by Aspergillus remains rare. We report the clinical and postmortem findings of an unusual case of Aspergillus pancarditis in a 7-year-old bone marrow transplant patient with Aspergillus embolization to the coronary arteries leading to a massive acute myocardial infarction. This case suggests that myocardial injury secondary to disseminated aspergillosis should be included in the differential diagnosis of chest pain in the immunocompromised pediatric patient. PMID- 3048490 TI - Specificity of monoclonal antibody Campath-1. PMID- 3048491 TI - Immunological reconstitution after bone marrow transplantation with Campath-1 treated bone marrow. PMID- 3048492 TI - T cell depletion in bone marrow transplantation. AB - Prior to the introduction of T cell depletion graft-versus-host disease (GVHD) was the major cause of transplant-related mortality. Why has the remarkable technical achievement of efficient T cell depletion, which has virtually eliminated severe GVHD over the last few years, failed to reduce mortality and increase disease-free survival? This review examines the value of T cell depletion based on recent clinical experience. It surveys the directions that are being pursued in an effort to solve the problems that have arisen and indicates how this is leading to a greater understanding of the mechanisms involved in GVHD and the graft-versus-leukaemia effect. PMID- 3048493 TI - Allogeneic bone marrow transplantation for leukemia: factors of importance for long-term survival and relapse. AB - From 1977 until June 1987, 139 patients with leukemia underwent allogeneic bone marrow transplantation at Huddinge Hospital. Among recipients of HLA identical bone marrow the survival plateau for 30 patients with acute myeloid leukemia in first remission was 71% from 3 to 7 years. Children (n = 11) had a survival of 100% compared with 52% (n = 19) in adults (p = 0.01). Among patients with acute lymphoblastic leukemia in first remission (n = 15) the actuarial 5-year survival was 73% compared with 31% for those (n = 25) in second to fourth remissions. The probability of relapse in these two groups was 9% and 45% respectively (p less than 0.05). Patients with chronic myeloid leukemia in first chronic phase (n = 27) had a survival plateau from 1 to 6 years of 65%. In Cox's multivariate regression analysis, improved survival was associated with grade 0-I acute graft versus-host disease (GVHD) (p less than 0.0001), HLA-matched siblings (p less than 0.001), recipient age less than 17 years (p = 0.02), first remission and first chronic phase (p = 0.03), cytomegalovirus (CMV) seronegative recipients (p = 0.04) and absence of symptomatic CMV infection (p = 0.04). A decreased risk of relapse was seen in patients with first remission or first chronic phase (p less than 0.001) and in patients with asymptomatic CMV infection (p = 0.03). In multivariate analysis the p values were 0.002 and 0.09 for these two groups respectively. In these patients acute GVHD was the major obstacle to successful outcome. PMID- 3048494 TI - Endocrine function after bone marrow transplantation without the use of preparative total body irradiation. AB - Ten children who underwent allogeneic (n = 5) or autologous (n = 5) bone marrow transplantation (BMT) for chronic myelogenous leukaemia (n = 2), acute lymphoblastic leukaemia (n = 1), acute myelogenous leukaemia (n = 2), severe aplastic anaemia (n = 2), malignant histiocytosis (n = 1), neuroblastoma (n = 1) and teratoma (n = 1) were assessed for endocrinological function. Transplant preparative regimens consisted of high-dose cyclophosphamide, high-dose cyclophosphamide in combination with high-dose busulphan, high-dose melphalan as well as BACT (BCNU, cytarabine, cyclophosphamide and 6-thioguanine) chemotherapy. None of the patients received total body irradiation (TBI). Median survival following BMT was 37 months (range 7-115). Growth hormone deficiency was present in only one patient; none of the patients had abnormal thyroid or adrenocortical function. This is in contrast to previous reports in which growth hormone deficiency and abnormal thyroid and adrenocortical function occurred in a much higher percentage of patients after BMT conditioned with TBI. PMID- 3048496 TI - Bone marrow transplantation for adults and children with poor risk acute lymphoblastic leukaemia in first complete remission. AB - Thirty-two patients with poor risk acute lymphoblastic leukaemia in first complete remission received bone marrow transplants (BMT) from fully matched family donors. Their ages ranged from 7 to 41 (median 23) years. Nine patients were aged 16 years or less. Patients were selected for BMT because they had risk factors for relapse with standard treatment approaches. In particular the children who were selected for BMT had presenting blast counts of greater than or equal to 90 x 10(9)/l or null immunophenotypes. The overall disease-free survival was 50% with a relapse risk of 31% at 9 years. Patients aged less than 16 years had a much lower risk of both graft-related disease and relapse than did older patients (disease-free survival 89% for those aged 7-16, 48% for those aged 17 26, 24% for those aged 26-41). We conclude that selection of BMT for young patients with poor risk features is entirely justified but that the prognosis for older patients is poor even after BMT. PMID- 3048495 TI - In vitro and in vivo cytokine-induced facilitation of immunohematopoietic reconstitution in mice undergoing bone marrow transplantation. AB - The aim of this study was to test whether colony stimulating factors (CSF) and other cytokines facilitate the recovery of a variety of immunohematopoietic functions in lethally irradiated mice undergoing bone marrow transplantation (BMT). Two experimental systems were employed: (a) lethally irradiated mice transplanted with syngeneic or T cell-depleted semi-allogeneic bone marrow (BM) cells (0.1-10 x 10(6)), subsequently treated by multiple doses of cytokines; and (b) lethally irradiated mice transplanted with BM cells that had previously been cultivated with cytokines. The cytokines used were: pure natural mouse interleukin-3 (IL-3); recombinant mouse granulocyte-macrophage CSF (rGM-CSF); recombinant human interleukin-2 (rIL-2); and crude cytokine preparations obtained from the culture supernatants of murine leukemia WEHI-3b cells (containing mainly IL-3), and of phorbol myristate acetate (PMA)-stimulated EL4 leukemia cells and concanavalin A-stimulated rat splenocytes (each containing a multitude of cytokines). For BM cultures (1-9 days), the cytokines were used at a dosage of 1 100 U/ml; for in vivo treatment, 2 x 10(2)-5 x 10(4) units were administered intraperitoneally and subcutaneously at different schedules for varying periods (1-3 weeks). The following parameters were tested 1-10 weeks post-BMT: white blood cell count, colony formation in agar and in the spleen of lethally irradiated mice, proliferative responses to mitogens and alloantigens, allocytotoxicity and antibody production (serum agglutinins and plaque-forming cells) against sheep red blood cells. Under appropriate conditions, cytokine treatment either in vitro or in vivo significantly enhanced (2- to 50-fold compared with controls) most functions tested at 2-8 weeks post-BMT, and shortened the time interval required for full immunohematopoietic recovery by 2-5 weeks. In recipients of semi-allogeneic, T lymphocyte-depleted BM no evidence of graft-versus-host disease was found. It is suggested that judicious application in vitro and/or in vivo of certain pure cytokines (e.g. GM-CSF, IL-3) or cytokine 'cocktails' might be beneficial in enhancing hematopoiesis and in the treatment of immunodeficiency associated with BMT. PMID- 3048497 TI - Safe application of a 13-Gy split dose total body irradiation schedule prior to bone marrow transplantation. AB - Two conditioning regimens for bone marrow transplantation for patients with acute lymphoblastic leukaemia were compared. Both regimens (VVRAPID and VVRAPID-X) incorporated the same cytotoxic chemotherapy but differed in the radiation dose; VVRAPID employed a single fraction of 10.5 Gy and VVRAPID-X employed 10.5 Gy followed 4 days later by a further dose of 2.5 Gy. The 13-Gy split-fraction schedule was well tolerated with no significant increase in infection or requirement for blood products. An increase in gastrointestinal toxicity occurred but responded to increased oral anti-diarrhoeal agents. Overall survival and relapse rates did not differ significantly between the two groups. There was a trend for the 13-Gy schedule to reduce the graft failure rate following T cell depleted bone marrow transplants. PMID- 3048499 TI - Marrow transplantation following busulfan and cyclophosphamide in multiple myeloma. AB - We report the use of busulfan and cyclophosphamide as conditioning before allogeneic bone marrow transplantation for a 38-year-old man with multiple myeloma. Three months post-transplant the monoclonal IgA band previously present was no longer detectable in the serum and plasma cells were no longer visible in the marrow. The patient died on day 93 with fungal infection. We conclude that the combination of busulfan and cyclophosphamide could be valuable before transplant for other patients with multiple myeloma. PMID- 3048498 TI - Renal involvement in chronic graft-versus-host disease: a report of two cases. AB - We report details of renal involvement during the course of chronic graft-versus host disease (cGVHD) in two patients undergoing bone marrow transplantation as treatment for acute leukemia. In both cases, the clinical picture was primarily characterized by proteinuria without hypertension or renal failure. Electron microscopy of renal biopsy specimens revealed a similar pattern in the two cases with extensive coalescence of foot processes and intramembraneous deposition of electron-dense material. Our data suggest that the kidney may be a target organ in chronic GVHD. PMID- 3048501 TI - Hypnosis as a window on regression. PMID- 3048500 TI - Busulphan and cyclophosphamide for pretransplant conditioning. PMID- 3048502 TI - A review of research supporting a dualistic view of mental operations. PMID- 3048503 TI - Pharmacotherapy and the neurobehavioural sequelae of traumatic brain injury. AB - Recent advances in clinical neuropharmacology are likely to improve the treatment and rehabilitation of traumatic brain injury (TBI) patients. Treatment may be directed to alleviate specific symptoms, to improve function in certain areas, or even to enhance the cortical recovery process. The author reviews pertinent issues in clinical neuropharmacology for the following drug classes: stimulants, other dopamine agonists, antidepressants, lithium, cholinergics, neuroleptics, anticonvulsants, beta-blockers, calcium channel blockers, nootropes, opiates and neuropeptides. Since the relevant research literature in TBI is so sparse, information and recommendations are extrapolated from some other patient groups, especially developmentally handicapped children and adults, and patients with dementia. PMID- 3048504 TI - Response to short-course chemotherapy of patients with initial resistance to antituberculosis drugs. PMID- 3048506 TI - A review of Campylobacter pylori in upper gastrointestinal disease. AB - The organism Campylobacter pylori is frequently found in association with gastritis and peptic ulcer disease. Whether it is the cause, a contributory factor or a simple commensal is not known but there is evidence to support a pathological role. Current research may alter our understanding of the causes of upper gastrointestinal disease and have important implications for treatment. PMID- 3048505 TI - HIV and the nervous system. AB - Neurological manifestations are common in HIV-infected individuals. Autopsy data suggest that the brain may be affected in as many as 80 per cent of cases. While opportunistic infections represent a major cause of neurological morbidity and mortality, there is growing evidence that HIV is itself neurotropic and can directly affect the nervous system. PMID- 3048508 TI - HLA still rules in renal transplants. PMID- 3048507 TI - The epidemiology of schizophrenia. AB - Epidemiology studies the factors affecting the frequency and distribution of diseases in populations. Surveys of the rates of schizophrenia, by studying the distribution of the disorder in populations, assist in the planning of services and provide clues about the aetiology. This article reviews the rates of schizophrenia in various countries, and the factors affecting its distribution. PMID- 3048509 TI - Stapling v. sutures for anastomoses. PMID- 3048510 TI - The management of massive haemorrhage. AB - Determined appropriate treatment of massive haemorrhage can reduce complications and save lives, yet failure to recognize or rectify acute blood loss undoubtedly still contributes to preventable hospital mortality. Anticipation and organization are the keys to improvement. PMID- 3048511 TI - What is a laser and how is it applied for therapy? AB - Lasers are now established as a major contributor to modern treatment techniques in a variety of disciplines. They have reached this position by providing improved precision, significantly less invasive surgical procedures and, in some cases, unique treatment modalities. Research into further applications continues to flourish ensuring that lasers retain their important role in medicine. PMID- 3048512 TI - Anaesthesia for elective abdominal aortic surgery. AB - This paper presents the purely personal attempts of one anaesthetist in a district general hospital to achieve a compromise from the widely varying advice given in print on the management of anaesthetics for elective abdominal aortic surgery. PMID- 3048513 TI - Magnetic resonance imaging of the abdomen--1988. AB - This article reviews the current clinical use of magnetic resonance imaging (MRI) in the abdomen with regard to recent technological advances, organ analysis and the integration of MRI in diagnostic imaging. The authors conclude that while MRI is not at present establishing itself as a leading diagnostic imaging method in the abdomen, if MRI machines were to become more widely available, abdominal applications could become a reality. PMID- 3048514 TI - Organizing organ donation. AB - Organ donation for transplantation involves intensive care staff in extra time and work. However, the results of renal, cardiac and hepatic transplantation are now so good (over 80% success) that every effort should be made to refer all suitable organ donors to the local transplant unit. PMID- 3048515 TI - Automatic implantable defibrillators. AB - The automatic implantable defibrillator was developed in the USA by Mirowski. This device automatically detects ventricular fibrillation or ventricular tachycardia and delivers a low energy (30 J) shock directly to the myocardium via patch electrodes on the epicardial surface or through a combination of a pericardial patch and an endocardial wire. These devices are being implanted in large numbers in the USA and represent a major contribution to the management and refractory ventricular arrhythmias. Unfortunately their high cost has limited their use in this country despite impressive clinical results. PMID- 3048516 TI - Diagnosis of factitious hypoglycaemia. PMID- 3048517 TI - Community care. PMID- 3048518 TI - The need to compare the effectiveness of the available antidepressant drugs. PMID- 3048519 TI - A defect in training. PMID- 3048520 TI - The neuropsychiatry of post-traumatic stress disorder. AB - The publication of DSM-III introduced the diagnosis Post-Traumatic Stress Disorder (PTSD), thus providing, for the first time, a framework for studying the consequences of extremely stressful events. Previously, traumatic neuroses had attracted a wide variety of labels - as wide as the experiences that produced them. Competing explanations in psychological and biological terms have characterised the approach to these disorders, and social and legal issues have added to the confusion. In recent years, psychosocial issues have tended to dominate the literature in relation to PTSD. While acknowledging the importance of such phenomenological and psychosocial approaches, this paper seeks to redress the balance by focusing on a biological perspective. PMID- 3048521 TI - Assaults on staff by psychiatric in-patients. A critical review. AB - Recent surveys indicate that the frequency of assaults on psychiatric staff by in patients has increased substantially over the past decade. With patient assaultiveness becoming of greater concern, the need to predict, anticipate, and avoid confrontations with potentially assaultive patients has grown. Regrettably, the vast majority of studies examining the nature, correlates, and predictors of in-patient assaultiveness suffer from methodological problems that limit the validity of their findings. PMID- 3048522 TI - The Geriatric Mental State Examination as a case-finding instrument in the community. AB - The Geriatric Mental State Examination (GMS), a standardised psychiatric interview, and its computerised diagnostic system, AGECAT, have been applied to a large (1070) sample of subjects aged over 65 in Liverpool. In a split-half study of this sample, diagnostic scales within the GMS and a diagnostic index derived from them were tested for efficacy in cases of organic and early organic illness (mostly various stages of dementia) and depressive illness. Results demonstrated high levels of sensitivity and specificity, and positive predictive values were within the expected range when applied to these disorders, given their low prevalence in the community. The GMS, when used in this way, is a flexible and effective case-finding instrument. PMID- 3048523 TI - Therapist contact and outcome of self-exposure treatment for phobias. A controlled study. AB - Eighty-four chronic phobic patients were randomly assigned to self-exposure in vivo instructed by either a psychiatrist, a computer or a book; mean therapy time per patient was respectively 3.1, 3.2 and 0 hours. Seventy-one patients completed treatment. All three groups improved substantially and similarly to 6 months follow-up, with no significant difference between them; self-exposure treatment was effective even without therapist contact. Among the three groups, initial expectation of help and positive attitude to the psychiatrist were equally high and related to subsequent rating of help received. All three groups rated the psychiatrist as more tolerant, reliable, and understanding than the computer or book, but these attitudes did not relate to outcome, were initially similar among all three groups, and changed minimally at 6 months follow-up. PMID- 3048524 TI - Biochemical aspects of therapy-resistant depression. AB - Despite the relatively high incidence of therapy-resistant depression, there is little clinical evidence to suggest that there are significant biochemical differences between therapy-resistant and therapy-responsive patients. A major problem for investigators is the lack of an internationally recognised definition of resistant depression. Also, it is possible that depressed patients with delusional symptoms or rapidly cycling affective disorders from subgroups that are more likely to be resistant to tricyclic antidepressant treatment and are therefore classified as therapy-resistant. Evidence is presented to show that an abnormality in serotonergic function may characterise some therapy-resistant depressed patients. PMID- 3048525 TI - Childhood and adolescent depression. I. Epidemiological and aetiological aspects. AB - Depression in childhood and adolescence has become a topic of considerable research interest in the last decade. A number of studies ranging over the last half-century provide information about the prevalence of depressive symptoms and syndromes in non-referred populations. These studies are critically reviewed in the light of an analysis of the various meanings that the term 'depression' may carry, and a variety of methodological issues. The sparse evidence for the involvement of a number of potential risk factors for depressive disorders is then considered and suggestions for future work in this area are outlined. PMID- 3048526 TI - Evaluation of treatment effectiveness in psychiatric research. PMID- 3048527 TI - Acetylcholine and Alzheimer's disease. AB - The hypothesis that cognitive impairment in Alzheimer's disease is related to cholinergic degeneration in the brain is still, a decade after its formulation, subject to critical evaluation. In marked contrast to the monoamine hypotheses of affective disorders or schizophrenia--based primarily on the mechanisms of action of therapeutic drugs, and yet lacking convincing pathological data on the human brain itself--the cholinergic hypothesis of Alzheimer's disease currently rests largely on evidence of neurochemical pathology in affected tissue, but still depends on effective therapy for its ultimate validation. The urgent need for a means of countering cognitive impairment in degenerative dementias such as Alzheimer's disease (probably the most important cause of intellectual decline in old age) hardly needs emphasising. In this annotation, a number of key questions specifically relating to the cholinergic involvement in Alzheimer's disease are considered. These questions are already being answered both within and, as so often in the history of biological psychiatry, outside the immediate area of investigation. PMID- 3048528 TI - Mental handicap: update. PMID- 3048529 TI - Issues of representation and limited capacity in the visuospatial sketchpad. AB - This article is about a visual short-term store called the visuospatial sketchpad (VSSP), which is tested by immediate ordered recall. Several claims are made: first, even for categorizable stimuli such as digits and letters, the representation in VSSP is uncategorized and unparsed. Second, the representation in VSSP is parsed and categorized just prior to recall in a process called recovery. The probability of recovering an item depends upon the fidelity of that item's representation. Third, VSSP has a fixed amount of representational medium which is allocated proportionally to the items in the presentation. When too many items are presented for recall, not all of the items are represented with sufficient fidelity to be recovered, producing the phenomenon of limited capacity. PMID- 3048530 TI - A serious complication of the Angelchik prosthesis. PMID- 3048531 TI - Femorodistal vein bypass graft stenoses. AB - Eighty femorodistal in situ vein bypass grafts have been evaluated at 3-monthly intervals clinically, with ankle:brachial pressure indices (ABI), by intravenous digital subtraction angiography (IV DSA) and by Duplex scanning. Five grafts (6 per cent) failed in the perioperative period. Nineteen (25 per cent) of the remaining 75 grafts subsequently developed stenoses on IV DSA during the first 12 months. All angiographic stenoses were detected by Duplex scanning using velocity ratio criteria before the development of symptoms or a measurable decline in ABI, i.e. while non-haemodynamically significant. This technique involves scanning the entire length of the graft but allows even minor stenoses to be detected and progression of stenoses can be determined. At a mean follow-up of 12 months (3-18 months), four (7 per cent) of the fifty-six grafts without stenoses occluded. Eight (42 per cent) of the nineteen stenosed grafts either occluded or developed symptoms. None of the occluded grafts in this series could be salvaged. Non haemodynamically significant vein stenoses can be detected non-invasively, occur frequently and are associated with graft failure. PMID- 3048532 TI - Human gastric perfusion: evidence for non-uniformity of blood flow. AB - Post-mortem injection studies have demonstrated marked differences in the course and distribution of arterioles in the stomach, but evidence of differential perfusion in man is lacking. Using the non-invasive laser Doppler technique, we studied 38 patients referred for gastroscopy for dyspeptic symptoms. Flux measurements were made at nine sites: distal oesophagus, cardia, mid-body and antrum on lesser and greater curves, and pre- and post-pylorus. In all cases the stomach was macroscopically normal. Flux in the proximal stomach was significantly greater than that in the antrum, (P less than 0.001, Student's t test). This in vivo study demonstrates a flow gradient related to the previously demonstrated anatomical differences. PMID- 3048534 TI - Use of older patients as cadaveric kidney donors. AB - We have no fixed upper age limit for cadaveric kidney donors and donors over the age of 50 provided kidneys for 22 per cent of our adult transplant recipients between 1983 and 1986. Immediate function following transplantation occurred in 17 per cent of these kidneys compared with 58 per cent for kidneys from donors under the age of 50. The 1-year actuarial graft survival rate for transplants from donors over 50 was 52 per cent, compared with 70 per cent for transplants from donors under 50 (P less than 0.05). Thus kidneys from older donors make an important contribution to the total pool of organs available for transplantation, but their use leads to inferior results in comparison with kidneys from younger donors. PMID- 3048533 TI - Trial of sclerosing agents in patients with oesophageal varices. AB - Forty-five cirrhotic patients with oesophageal varices underwent endoscopic injection sclerotherapy in a prospective randomized trial carried out to compare two sclerosing agents (5 per cent ethanolamine oleate and 2 per cent sodium tetradecyl sulphate (STD] with respect to safety, efficacy and complications. Twenty-three patients were allocated to the ethanolamine group and twenty-two to the STD group. The rate of control of acute bleeding was 100 per cent (6/6) in the ethanolamine group and 75 per cent (3/4) in the STD group. There was a significantly lower rate of postinjection bleeding after the over-tube was removed at the initial session of sclerotherapy when ethanolamine was injected 0/23 versus 7/22, 32 per cent; P less than 0.01) and at the second session there was a significantly (P less than 0.01) higher rate of jet-like bleeding from injection sites in the STD group (6/21, 29 per cent) than in the ethanolamine group (0/22). The disappearance rate of red colour signs 1 week after the initial session of sclerotherapy in the ethanolamine group was 100 per cent and 62 per cent in the STD group. Early oesophageal ulcers developed less frequently in the ethanolamine group (0 and 9 per cent) than in the STD group (24 per cent and 43 per cent both after the initial (P less than 0.05) and the second session of sclerotherapy (P less than 0.01]. Early bleeding from an oesophageal ulcer occurred only in the STD group (5/21, 24 per cent) before the third session of sclerotherapy (P less than 0.05). The rate of early mortality did not differ between the two groups. We conclude that ethanolamine seems to be safer and more efficacious than STD for sclerosing oesophageal varices. PMID- 3048535 TI - Application of mammary serum antigen assay in the management of breast cancer: a preliminary report. AB - A monoclonal antibody (3E1.2) based serum test using an enzyme immunoassay has been used to determine circulating levels of the breast cancer associated antigen -mammary serum antigen (MSA). Of 157 patients with early breast cancer (stage I and II) and 199 patients with advanced breast cancer (stage III and IV), 73 per cent and 87 per cent respectively had elevated MSA levels (i.e. greater than 300 inhibition units (IU). Furthermore, 40 of 44 patients (91 per cent) had a significant fall of MSA levels with reduction in tumour load by mastectomy. In addition, there was a correlation of MSA levels with the clinical course: changes in MSA levels correlated with changes in disease status (progressive disease, stable disease, disease regression) in 54 of 61 patients and antedated disease progression or recurrence by up to 8 months in some patients; and in 32 of 36 patients (89 per cent) with no clinical evidence of recurrence MSA levels did not vary by more than 25 per cent of the original MSA value over a period of 2-15 months. MSA is therefore a useful tumour marker in the diagnosis and staging of breast cancer. There is also evidence that serial estimations of MSA levels may be used to detect subclinical recurrence and the fluctuations in MSA levels might be useful in assessing response to therapy. Furthermore, it was also noted that surgical procedures such as fine needle aspiration biopsy or incisional biopsy could lead to a rise in MSA levels. PMID- 3048536 TI - Anaesthetic management of small animal patients with endocrine disease. PMID- 3048537 TI - The colonization of the alimentary tract and visceral organs of chicks with salmonellas following challenge via the feed: bacteriological findings. PMID- 3048539 TI - Effects of estrogen on hypothalamic gonadotropin-releasing hormone release in castrated male rhesus macaques. AB - The evidence that hypothalamic gonadotropin-releasing hormone (GnRH) release increases during the estrogen-induced luteinizing hormone (LH) surge in castrated female macaques and that estrogen induces a similar LH surge in the male suggests that either sexual differentiation of GnRH secretion does not exist or that changes in brain GnRH are not critical for ovulation. We tested this hypothesis by using the push-pull perfusion (PPP) technique to monitor GnRH release in the mediobasal hypothalamus continuously from 10 h before to 50 h after estrogen injection in 9 castrated male rhesus macaques. Blood sampling and PPP were performed with monkeys freely moving in their own cage by using a specially designed swivel/tethering system. PPP samples were collected every 10 min and analyzed for GnRH concentration by radioimmunoassay. Blood samples were collected every hour and plasma LH was measured by bioassay. Estradiol benzoate (EB, 42 micrograms/kg, b. wt.) was subcutaneously injected after 10 h of initial PPP. The PPP was continued for 10 h after EB in 4 and 50 h after EB in 5 monkeys. Hypothalamic GnRH patterns were analyzed by the Pulsar algorithms. The results show that during the 10 h after EB, plasma LH declined rapidly, reaching low or non-detectable levels by 7-9 h, while hypothalamic GnRH releasing patterns did not change during this period (n = 9). In contrast, estrogen enhanced GnRH level and pulse amplitude, but not pulse frequency, several hours before mean peripheral plasma LH increased from suppressed values (n = 4).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3048538 TI - Neurotoxicity of N-methyl-D-aspartate is markedly enhanced in developing rat central nervous system. AB - The neurotoxic lesion produced by direct injection of 25 nmol of N-methyl-D aspartate (NMDA) into the corpus striatum of 7-day-old rats was compared to the effects of injecting 75 nmol into the striatum or hippocampus of adults. The area of histopathology in the immature striatum was 21 X larger than the striatal lesion in adults. Damage from NMDA injected into the immature striatum also extended into the dorsal hippocampus and produced an area of destruction which was 16 X larger than observed after direct injection into the adult hippocampus. Several studies have implicated excessive N-methyl-D-aspartate receptor activation in the pathogenesis of hypoxic-ischemic and hypoglycemic injury and our results suggest that this neurotoxic mechanism is extremely active in the immature brain. PMID- 3048540 TI - [The means (and possible reinforcement) of civil security]. PMID- 3048541 TI - [The medical chain during medical assistance in catastrophes]. PMID- 3048542 TI - [Intervention detachments]. PMID- 3048544 TI - [Smoking today and physicians]. PMID- 3048543 TI - [Principles of triage applied in wartime and in catastrophes]. PMID- 3048545 TI - [The extension of deep venous thrombosis treated by intravenous heparin: apropos of 12 cases]. PMID- 3048546 TI - [Does radioisotope methodology justify the existence of acupuncture meridians?]. PMID- 3048547 TI - [Identification by immuno-transfer of probable immunogenic fractions extracted from exo-antigens of Plasmodium falciparum]. PMID- 3048548 TI - [Characterization of the subtype of purinergic receptor involved in the stimulation of insulin secretion]. PMID- 3048549 TI - [Microcirculatory aspects of Reilly's phenomena]. PMID- 3048550 TI - [Eulogy of Julien Marie (1899-1987)]. PMID- 3048551 TI - [Influenza vaccination: reinforcement of its effectiveness by combination with an immunostimulant]. PMID- 3048553 TI - [Human experimentation]. PMID- 3048552 TI - [Authorization request for the use of L-carnitine, calcium chloride and magnesium sulfate as nutritional additives in the formulation of food designed for specialized diets]. PMID- 3048554 TI - [The nomenclature of professional services]. PMID- 3048555 TI - [Hereditary lipidosis with triglyceride overload: a metabolic study in cultured cells]. PMID- 3048557 TI - [Anti-venereal dispensaries, the campaign against sexually transmitted diseases and their prevention]. PMID- 3048556 TI - [Essential varices of the internal saphenous vein]. PMID- 3048558 TI - [Eulogy of Jean Delay (1907-1987)]. PMID- 3048559 TI - [Air pollution in indoor locations]. PMID- 3048560 TI - [Substances used in animal production]. PMID- 3048562 TI - [The treachery of words: apropos of "beyond coma"]. PMID- 3048561 TI - [Eulogy of Robert Fasquelle (1908-1987)]. PMID- 3048563 TI - [The public assistance day care hospital in Paris]. PMID- 3048564 TI - Aeromedical transfer of burned patients: a review with special reference to European civilian practice. AB - Inter-hospital transfer of a patient with a burn injury can usually be undertaken with confidence, even for non-medical reasons, as long as it is undertaken with care. Transfer by air has many advantages for the seriously burned patient provided that all potential risk factors are taken into account. There is a need to consider infection, the respiratory condition and the cardiovascular status of the patient. The possibility of other injuries should also be borne in mind. Early transfer requires careful but urgent planning if it is to be achieved, as is desirable, by days 3 or 4 postburn. Transfer during or at the end of the resuscitation phase, or in the convalescent phase when skin cover is well established, poses least problems. Plenty of sterile paraffin gauze, wool and crepe bandages will be needed. Appropriate monitoring equipment and full cardiovascular and respiratory resuscitation facilities should be carried. Care should be taken in the selection of the medical and nursing personnel to accompany the patient, who should whenever possible have experience of burns as well as aviation medicine. PMID- 3048566 TI - Unexplained blindness after a major burn. AB - A 43-year-old woman experienced the sudden onset of unexplained bilateral blindness 42 days after a 55 per cent total body surface area burn. Possible causes are examined, and a comparison with cases previously reported in the literature is made. PMID- 3048565 TI - Burn wound carcinoma: case report and review of the literature. AB - A 22-year-old man developed squamous cell carcinoma of the exophytic papillary type following a latent period of 14 years in a burn scar overlying the left forearm and wrist, with concomitant left axillary lymph node metastasis. The literature pertaining to clinical presentation, pathological findings and prognosis of such patients has been reviewed. The surgical treatment and adjuvant therapy have been discussed. Patients should be counselled to seek medical advice early for persistent ulcerations or deep scars of burn wounds, especially those overlying joint surfaces. PMID- 3048567 TI - Burns during pregnancy. AB - Eight cases of burns during pregnancy were treated in Kuwait during the last 3 years. In all burns above 30 per cent body surface area (BSA), abortion, premature labour and intrauterine fetal death are ever-present complications and the survival of the foetus in burns above 50 per cent BSA is uncommon. Nearly all these complications occur during the first week after the burn. For patients in the second and third trimesters of pregnancy with burns above 50 per cent BSA the prognosis is usually poor unless the pregnancy is terminated within 24-48 h after the burn. PMID- 3048569 TI - FUGE: an analytical ultracentrifuge simulation for teaching purposes. AB - An IBM-compatible microcomputer program for teaching purposes is described which stimulates the operation of a sedimentation velocity determination of a protein in an analytical ultracentrifuge using schlieren optics. The program operates in speeded-up time and simulates the major procedures which would need to be carried out to operate such an instrument. The position of the sedimenting boundary can be observed at any time during the run, and up to six 'photographs' can be recorded for subsequent analysis. Calculation of sedimentation coefficient, diffusion coefficient and mol. wt can be made from a dot-matrix printout. Ten representative proteins are stored within the program, but provision exists for user-supplied data. PMID- 3048568 TI - The mesh skin graft--true expansion rate. AB - Using the mesh graft II dermatome (Zimmer) for split skin graft expansion, the 3 to 1 and 1.5 to 1 expansion rates were evaluated for true clinical expansion. In one hundred and one 1.5 to 1 expanded grafts the actual expansion was 1.2 and in sixty 3 to 1 expanded grafts, it was 1.5. PMID- 3048570 TI - Homeopathy in Illinois. PMID- 3048571 TI - Joseph-Frederic-Benoit Charriere (1803-1876): Paris surgical instrument maker from Switzerland. PMID- 3048572 TI - Charriere's instruments in America. PMID- 3048573 TI - The history of the health sciences at the National Museum of American History. PMID- 3048574 TI - Attempted prevention of blast crisis in chronic myeloid leukemia by the use of pulsed doses of cytosine arabinoside and cis-chloronitrosurea during the course of busulfan-maintained remission. AB - We performed this chemotherapeutic trial to try to delay the onset of the blast crisis of chronic myeloid leukemia (CML) by pulsing doses of drugs most likely to be effective against emerging "blast" cells characteristic of acute phase disease. A randomized trial in patients with CML comparing busulfan maintenance to busulfan maintenance plus pulsed doses of cytarabine and lomustine did not yield any differences in either time to blast crisis or death. PMID- 3048575 TI - In vitro production of beta 2 microglobulin by human myeloma cells. AB - Serum beta 2 microglobulin (B2M) has recently been shown to be a powerful, although nonspecific, marker of multiple myeloma (MM) disease activity. Since the nature of cells producing high levels of B2M remains obscure in MM, we measured (in vitro) the spontaneous secretion of free B2M in 58 culture samples from 52 patients with MM. In comparison with samples from normal individuals and from individuals with monoclonal gammopathy of unknown significance, an abnormal secretion of B2M was found in 83% of samples containing myeloma cells. Levels of secretion significantly correlated with the percentage of tumor cells, tumor progression, and moreover, with the immunoglobulin type of the tumor. The highest levels of secretion were noted in IgG and IgA MM. Our present results would favor the hypothesis of a direct secretion of B2M by myeloma cells and emphasize the interest of B2M as a marker in a majority of (but not all) patients with MM. PMID- 3048576 TI - Reactivity of the monoclonal antibody B72.3 with fetal antigen: correlation with expression of TAG-72 in human carcinomas. AB - The monoclonal antibody (MAb) B72.3 recognizes a mucin-like glycoprotein, TAG-72, which has been detected in a spectrum of human carcinomas, but not in the normal tissue counterparts. Using avidin-biotin-peroxidase complex (ABC) immunohistochemical techniques, MAb B72.3 was reacted with formalin-fixed, paraffin-embedded fetal and pediatric tissue sections to determine the extent of expression of the recognized antigen in these tissues. First trimester fetal tissues failed to express detectable antigen. Gastrointestinal epithelia from 13 to 34 weeks gestation demonstrated the most immunoreactivity with B72.3 although bronchial respiratory epithelium of the lung, transitional epithelium from the kidney, Hassall's corpuscles of the thymus, and gonadal tissues from fetuses of both sexes were also reactive. The TAG-72 antigen was not detected in fetal breast, pancreas, liver, spleen, adrenal, or heart. Expression of the TAG-72 antigen in malignancy appears to correlate well with fetal tissue reactivity with B72.3. PMID- 3048577 TI - The development and management of chemotherapy-related anticipatory nausea and vomiting. PMID- 3048578 TI - Medicare payment for physician services: implications for specialists. PMID- 3048579 TI - Triangles of abuse: a model of maltreatment. AB - Research and clinical evidence support the view that the perpetrator (P), victim (V), and observer (O), all contribute to child abuse and neglect. This paper describes a model in which they have a necessary part in precipitating and sustaining the maltreatment. It appears they form a triangle or triquetra which rotates with time and circumstance. The perpetrator of sexual assault often becomes the victim when he is imprisoned. The victim of physical abuse may become the perpetrator of physical abuse in the next generation. Within each P, V, and O there is also a triquetra which rotates and which will determine how that individual behaves in the next instance. Knowing the relative preponderance of P, V, or O within each individual can help predict possible future abuse. Since therapists often determine their intervention by the view of the situation they adopt, it is important for them to understand their principal orientation towards either the perpetrator, victim, or observer. The legal model of abuse emphasizes individual accountability by viewing people as they are solely responsible for their behavior. This article rather than diminishing the responsibility of any individual, emphasizes the collective responsibility of all involved. PMID- 3048580 TI - The efficacy of hospitalization of nonorganic failure-to-thrive children: a meta analysis. AB - This study ascertained the efficacy of hospitalization of children with nonorganic failure to thrive (NOFTT). A meta-analysis was performed on previously published research findings which met certain criteria for inclusion in the analysis. Two categories of outcome measures, physical growth pattern and psychosocial development, were quantitatively synthesized separately using standard meta-analytic techniques. Hospitalization was found to significantly enhance the probability of sustained catch-up physical growth among NOFTT children, but only a comparatively small effect size for hospitalization on their psychosocial development was documented. There remains need for prospective longitudinal study of effectiveness in treating NOFTT children. But meta-analysis is a powerful technique which introduces scientific rigor to the review of study results, none of which is singularly conclusive. PMID- 3048582 TI - [Colonic pseudo-obstruction: pathogenesis and treatment]. PMID- 3048581 TI - [Tracheal access for jet ventilation in otorhinolaryngologic surgery]. PMID- 3048583 TI - [Stress reaction and anesthesia. The role of morphine agonists and antagonists]. PMID- 3048584 TI - Septic shock: a clinical perspective. AB - Septic shock may occur in otherwise normal individuals but is frequently a fatal sequel to infection in the elderly, the diabetic, or the debilitated patient. Mortality rates range from 40 to 95 per cent depending both on host factors and on the speed of initiation of appropriate therapy. The complexities of shock pathophysiology and biochemistry are elusive, but certain hemodynamic derangements are clearly described and create obvious therapeutic imperatives. Correction of the supply-dependent phase of oxygen consumption is the cornerstone of supportive therapy. Survival is primarily dependent on the rapid delivery of the appropriate antibiotics, surgical drainage and debridement of any infected tissues or abscesses, and aggressive volume resuscitation at the very time early sepsis is diagnosed. Septic shock is a medical emergency. PMID- 3048586 TI - Nosocomial pneumonia. AB - Nosocomial pneumonia remains a challenging problem in critically ill patients in terms of both diagnosis and therapy. The clinical picture is often confusing; confounding factors such as congestive heart failure, ARDS, and interstitial lung disease may obscure the presence of pneumonia. Previous antimicrobial therapy or the presence of large numbers of colonizing organisms contribute to the difficulty of diagnosis. The use of sheathed fiberoptic bronchoscopy with quantitative culture and biopsy is probably the best initial invasive test when routine diagnostic methods fail; open lung biopsy remains the ultimate standard for diagnosis. Empiric therapy is often necessary and should be designed to treat organisms suspected of being the etiologic pathogens either on the basis of preliminary laboratory results (gram and acid-fast stains) or the clinical setting. PMID- 3048585 TI - Post-infectious ARDS: mechanisms of lung injury and repair. AB - Lung infections are considered the most common causes of ARDS. Additionally, superimposed secondary lung infections may occur in the setting of ARDS and further aggravate lung damage. Substances released by neutrophils are the major mediators of lung injury, although other factors, such as endotoxin, exotoxins, macrophages, and immune mechanisms, are significant contributors. Neutrophil induced lung injury results from the production of oxygen radicals, the release of products of arachidonate metabolism, and the activation of lysosomal enzymes. The development of ARDS is associated with high mortality. Although a large proportion of survivors eventually recover adequate lung function, some patients progress to severe fibrosis. Experimental evidence suggests that the development of fibrosis results from increased synthesis of ECM components by lung fibroblasts. Fibroblast synthesis of ECM components is normally influenced by alveolar cells, by circulating growth factors, by the external milieu, and by the extent of gene expression of the specific ECM components. These factors undergo significant alterations during the evolution of experimental lung fibrosis. Growth factors are more abundant, alveolar cells are activated, and gene expression is enhanced. These factors, which play a crucial role in promoting experimental lung fibrosis, are likely to be involved in the pathogenesis of fibrosis in post-infectious ARDS. PMID- 3048587 TI - The febrile granulocytopenic patient in the intensive care unit. AB - Patients treated aggressively for potentially curable hematologic and neoplastic diseases are often admitted with profound granulocytopenia to an intensive care unit. These patients have a high risk of sustaining life-threatening infections caused by various bacterial, fungal, viral, and protozoal pathogens. Successful management of these critically ill, profoundly granulocytopenic patients by the intensive care team requires an organized, informed, and rational approach to treating their infections. PMID- 3048589 TI - Antibiotic selection and pharmacokinetics in the critically ill. AB - In critically ill patients, a good outcome of an infection episode often requires early institution of appropriate antimicrobial therapy. The choice of one specific antimicrobial agent or combination of agents over another requires consideration of the nature of the etiologic organism, the host, and the drug(s). PMID- 3048588 TI - Burn sepsis. AB - Sepsis in the burned individual can arise from multiple causes. However, the unique source is the burn wound itself. It is clear that health is association with maintenance of a bacterial equilibrium in the wound and that infection is a result of an imbalance in favor of the bacteria. The primary host defense mechanism, an intact epithelial barrier, has been lost at the time of burning. A portal of entry has been created, and the bactericidal defenses have been neutralized. All of the host defense mechanisms associated with inflammation are evoked but may be limited by the avascular isolation of much of the wound. In addition to alteration in vascular response associated with the burns, there are adverse changes in the neutrophils themselves. The alterations in nutrition that may follow burn injury further reduce systemic host resistance. Associated diseases, such as diabetes, may present a further hazard. All of the local factors influencing host resistance are adversely affected in the burn wound. There is necrotic tissue, decreased local tissue perfusion, and loss of the mechanical barrier. Quantitative techniques have demonstrated that bacteria are present in the depths of the wound from the time of injury. Infection and burn wound sepsis are clearly represented by the quantitative increase in bacteria to numbers exceeding 10(5) per gram of tissue. In no other instance has the importance of the "amphibiont" organisms been more clearly demonstrated than in the burn wound. Today's nonpathogen has all too often become tomorrow's killer. As therapeutic control becomes effective against the current organism, the ecologic void is filled by another, which, by definition, is resistant to the treatment being employed. PMID- 3048590 TI - Candida infections in the intensive care unit. AB - Infections caused by Candida species are now endemic and entrenched in the intensive care unit setting. Candida infections continue to provide a major diagnostic and therapeutic challenge to clinicians. The increased prevalence of Candida infections represent the high price clinicians and patients must pay for the advances in invasive diagnostic and therapeutic technology. This article deals with the epidemiology, pathogenesis, diagnosis, and management of common Candida infections in the critically ill. PMID- 3048591 TI - The septic multiple-trauma patient. AB - Sepsis in the multiple trauma patient is being seen with increased frequency now that more of these patients are surviving the initial period. Traumatic destruction of tissue barriers, the placement of various tubes and drains, and surgical repair with debridement all provide conduits for colonization and infection with pathogens. Many components of the host immune system also become altered after trauma and surgery, predisposing this population to infectious complications. The site of infection can be cryptic in the moribund trauma patient; locating it may require many special diagnostic procedures. Continuing close surveillance is important to prevent or to identify infections at the earliest possible time. The liberal use of antibiotics should be discouraged so that development of resistant organisms and superinfection is kept to a minimum. Handwashing between patient contacts may be the most important prophylaxis against the spread of pathogens within a trauma unit. PMID- 3048593 TI - Pulmonary diagnostic procedures in the critically ill. AB - When faced with a critically ill patient with new pulmonary infiltrates on chest roentgenograms, the physician must choose the appropriate diagnostic procedure on the basis of the expected yields versus the potential complications. The first steps in any patient should include discontinuation of any nonessential medications, careful evaluation of fluid status to exclude cardiogenic pulmonary edema, and a review of likely diagnoses based on the patient's underlying disease. Although not likely to be of immediate utility, obtaining cultures of blood and other body fluids or sites and serologic testing may provide helpful information when combined with other procedures. Bronchoscopy is a reasonable first step in patients with a slow progression of disease or in those in whom the pulmonary process is discovered early in its course. As these patients often present with several of the known risk factors for complications with bronchoscopy, the decision to perform this procedure should not be made lightly. Transbronchoscopic lung biopsy adds additional risk to bronchoscopy but also increases the diagnostic yield considerably over lavages, brushing, and bronchial washings. Open lung biopsy offers high diagnostic yields and relatively low rates of serious complications. Because of the invasive nature of the procedure, there is often reluctance to perform it. In patients with rapidly progressive disease or conditions that make the risk of bronchoscopy unacceptably high, such as severe hypoxemia, bleeding diathesis, or cardiac compromise, prompt diagnosis requires that the physician consider open lung biopsy as a first diagnostic procedure. The physician must also consider whether making a specific diagnosis will be of benefit to the patient. Potential benefits of a specific diagnosis include stopping unnecessary empirical (and potentially toxic) therapies, instituting correct and specific therapy, and thus decreasing morbidity and mortality. The impact of specific diagnosis on morbidity and survival is often difficult to demonstrate. Discouraging notes have been sounded by studies of the effect of bronchoscopic or surgical diagnosis on the ultimate outcome for patients. For bronchoscopy with transbronchoscopic lung biopsy, although the overall diagnostic rate was 60 per cent, no difference in survival was noted between patients in whom a diagnosis was made and those in whom the nature of the pulmonary process remained unknown. Similarly, in a series of patients who underwent open lung biopsy, although the results of biopsy led to a therapeutic change in 70 per cent of the patients, only 16.5 per cent of the patients benefited from this change.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3048592 TI - The management of fulminant meningitis in the intensive care unit. AB - Fulminant meningitis requires aggressive management in an intensive care unit setting. The pathophysiology of the various factors that damage the central nervous system in this disease have been reviewed, as well as the management of the many complications of this serious, often devastating, infection. The etiologic agents according to age group have been discussed, and recommendations for empiric therapy have been made. PMID- 3048594 TI - Canada's oldest dentist marks 80th year since his graduation (Dr. Clarence Brooks). PMID- 3048595 TI - Smokeless tobacco and oral disease. A review. PMID- 3048597 TI - A conservative cosmetic identification concept for prosthetic appliances. PMID- 3048596 TI - Effect of a daily 0.2% chlorhexidine rinse on the oral health of an institutionalized elderly population. PMID- 3048600 TI - Single lung transplant--experience with a canine model. PMID- 3048598 TI - [Crevicular fluid. I: Clinical significance]. PMID- 3048601 TI - Insulin binding in erythrocyte from patients with obese diabetics. PMID- 3048602 TI - [Refractoriness on repeated methacholine provocation test]. PMID- 3048599 TI - [Crevicular fluid. II: Collection]. PMID- 3048603 TI - Cervical chlamydial infection in gynecologic outpatients. PMID- 3048604 TI - Ganser syndrome--an unusual case report and literature review. PMID- 3048605 TI - [Measurement of liver volume by ultrasound]. PMID- 3048606 TI - [Small bowel perforation secondary to metastatic carcinoma of lung]. PMID- 3048607 TI - [Epidermoid carcinoma of the kidney]. PMID- 3048608 TI - Vaginal botryoid embryonal rhabdomyosarcoma (sarcoma botryoides), A report of one case and review of the literature. PMID- 3048609 TI - From variables to videodiscs. Interactive video in the clinical setting. PMID- 3048611 TI - 3-O-methyl-D-glucose uptake in cultured bovine adrenal chromaffin cells. AB - The membrane transport of glucose was studied in bovine adrenal chromaffin cell cultures by following the cell/medium distribution of the nonmetabolizable glucose analog, 3-O-methyl-D-glucose. Uptake of this sugar in day-1 cultures that are undergoing rapid morphological change and differentiation had a Vmax of 138 nmol/(mg protein.min) and Km of 15 mM, and was only slightly increased by 50 mU/mL insulin. In day-5 cultures where morphological changes were essentially completed, Vmax and Km decreased to 51 nmol/(mg protein.min) and 9.5 mM, respectively, and the response to insulin was restored to the level found in freshly isolated cells; this effect was abolished in the nominal absence of Ca2+. Thus, saturation kinetics and insulin and Ca2+ sensitivity of 3-methylglucose uptake observed in freshly isolated cells were maintained in culture. However, the insulin response was almost absent during the initial period of rapid morphological change when sugar transport was strongly stimulated. Culture of chromaffin cells in the presence of dexamethasone did not inhibit the formation of processes, but decreased 3-methylglucose uptake in day-5 cultures by an apparently competitive effect. PMID- 3048612 TI - Proprioceptors and their contribution to somatosensory mapping: complex messages require complex processing. AB - This review of other people's work concentrates on two matters. First, which of the various receptors are chiefly involved in creating the central representations or maps of the body that both underlie the conscious perception of our body image and are needed to control our motor performance. Second, how are these relatively well-charted signals used in sensorimotor mapping, with particular attention paid to various human psychophysical observations and illusions that throw light on the central integrative mechanisms involved. Detailed citation is largely restricted to developments since earlier reviews. PMID- 3048613 TI - The representation of arm movements in postcentral and parietal cortex. AB - Considerable experimental evidence supports the hypothesis that the neocortical processes underlying kinesthetic sensation form a hierarchical series of cells signalling increasingly complex patterns of movement of the body. However, this view has been criticized and the data lack quantitative verification under controlled conditions. These studies have also typically used one-dimensional (reciprocal) movements, even of multiple degree-of-freedom joints such as the wrist or shoulder, and have been restricted to passive movements. This latter limitation is particularly critical, since the response of many muscle receptors is affected by fusimotor activity while that of many articular receptors is sensitive to the level of muscle contractile activity. Both factors introduce significant kinesthetic ambiguity to the signals arising from these receptors during active movement. This ambiguity is evident in the discharge of primary somatosensory cortex proprioceptive cells. Studies in area 5 show that single cells signal shoulder joint movements in the form of broad directional tuning curves. The pattern of activity of the entire population encodes movement direction. The cells appear to encode spatial aspects of movement unambiguously, since their discharge is relatively insensitive to the changes in muscle activity required to produce the same movements under different load conditions. It is not yet certain whether the somesthetic activity in area 5 is a kinesthetic representation that is sequential to and hierarchically superior to that in SI, or whether it is a parallel representation with separate and distinct function. PMID- 3048610 TI - Rapid measurement of blood ethanol concentrations using the alcohol oxidase membrane technique. AB - The alcohol oxidase membrane technique is available for measurement of ethanol in commercial fluids. In this paper we examined its usefulness for cat and human blood in comparison with gas-liquid chromatography (GLC). The membrane method proved to be simple, reproducible, accurate, and inexpensive. Analysis took 1-2 min per sample and required only 25 microL of whole blood for measurement of concentrations between 0.05 and 1.0 mM (0.25-5 mg/dL) and 10 microL of whole blood for measurement of concentrations between 1.0 and 40 mM (5-190 mg/dL). Background concentrations were undetectable in cats after extraneous sources of alcohols were removed. The alcohol oxidase membrane technique is less specific than GLC, but it may be useful when ethanol is administered after background samples have shown an absence of other nonspecific reactants. Its high sensitivity is useful for kinetic studies where blood ethanol concentrations are below or close to those required for maximal hepatic ethanol metabolism. PMID- 3048615 TI - Representation of three-dimensional visual space in the cerebral cortex. AB - This article reviews two issues relevant to the topic of how three-dimensional space is represented in the cerebral cortex. The first is the question of how individual neurons encode information that might contribute to stereoscopic estimation of visual depth. Particular attention is given to the current understanding of the neural representation of motion through three-dimensional space and to the complexities that arise in interpreting neuronal responses to this complex stimulus parameter. The second issue considered is the disorderlines that exists in the retinotopic mapping of the visual field in some cortical visual areas. Several extrastriate areas have been found to contain maps of the contralateral visual hemifield that are disorderly in the sense that the representation of various parts of the visual field are often misplaced or grossly over-or under-represented. It is suggested that this disorderlines may in some cases represent adaptations to facilitate certain types of visual functions. PMID- 3048614 TI - Computational studies of the extraction of visual spatial information from binocular and motion cues. AB - This paper reviews some of the contributions that work in computational vision has made to the study of biological vision systems. We concentrate on two areas where there has been strong interaction between computational and experimental studies: the use of binocular stereo to recover the distances to surfaces in space, and the recovery of the three-dimensional shape of objects from relative motion in the image. With regard to stereo, we consider models proposed for solving the stereo correspondence problem, focussing on the way in which physical properties of the world constrain possible methods of solution. We also show how critical observations regarding human stereo vision have helped to shape these models. With regard to the recovery of structure from motion, we focus on how the constraint of object rigidity has been used in computational models of this process. PMID- 3048616 TI - Saccadic command signals in the superior colliculus: implications for sensorimotor transformations. AB - Recent findings concerning the anatomical and functional organization of the deep division of the primate superior colliculus are summarized. These data are interpreted as supporting the hypothesis that the deeper layers of the superior colliculus are organized in motor, rather than sensory, coordinates. According to this hypothesis, a dynamic remapping of sensory signals is required for interfacing with the map of motor error space contained in the superior colliculus. PMID- 3048617 TI - Comparative hemostatic protein alterations accompanying pregnancy and parturition. AB - Pregnancy and parturition represent adaptive, physiological stress situations that elicit both blood coagulation protein changes and endocrine alterations. This paper briefly reviews the interrelationships of these responses by comparing the available data in several mammalian species including man, dog, cow, and pig. Hemostasis is an enzymatically controlled cascade process involving platelets and specific coagulation proteins that results in the formation of insoluble strands of fibrin. Although qualitatively similar in most mammals, quantitative differences exist in the circulating levels of individual coagulation proteins which may contribute towards the different rates of clot formation. In the late gestation stage of human pregnancy, significant increases are observed in the circulating biological activities of the coagulation factors VII, VIII:C, IX, and X. In the dog, the activities of factors VII and IX reach peak values in midpregnancy, while in the cow little or no changes are observed except for factor VII values which show a significant increase only around the time of parturition. These coagulation changes appear to be associated with fluctuations in circulating hormone values, particularly plasma estrogen. In human pregnancy, the circulating levels of fibrinogen, the precursor of fibrin, increase during the gestation period reaching maximum values during and immediately following parturition. In the dog and the cow two distinct increases in fibrinogen values are observed, the first in the midgestation period and the second in the immediate postparturition period. In the pig, fibrinogen values also rise significantly at parturition. Oral contraceptive studies in the human female have indicated that changes in circulating activities of coagulation proteins are hormonally influenced.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3048618 TI - Human chorionic gonadotropin and luteinizing hormone-releasing hormone reverse the blockade of ovulation in pregnant mare's serum gonadotropin-primed immature rats by the anti-androgenic drug, hydroxyflutamide. AB - The present study was designed to examine mechanism(s) of the anti-ovulatory action of the anti-androgen, hydroxyflutamide (OH-F). Prepubertal rats were treated with 4 IU pregnant mare's serum gonadotropin (PMSG) (day -2) to induce first estrus and ovulation. They received OH-F in sesame oil or oil alone at 08:00 and 20:00 h on day 0 (the day of proestrus) and ovulations were assessed on the morning of day 1. Eighty-three percent of control animals ovulated with a mean of 7.7 +/- 1.1 corpora lutea per rat. Hydroxyflutamide blocked ovulation in all but 2 of the 12 rats receiving this drug alone. All of OH-F treated rats that received 5 and 25 IU human chorionic gonadotropin (hCG) ovulated with means +/- SEM of 9.1 +/- 0.1 and 7.3 +/- 1.4 corpora lutea per rat, respectively. The dose of 0.2 IU hCG was essentially ineffective, while the effect of 1.0 IU hCG was intermediate. At the dose of 20 ng and above (100 and 500 ng) luteining hormone releasing hormone (LHRH) completely overcame the ovulation blockade in the OH-F treated animals, while a 4-ng dose was ineffective. At 18:00 h on the day of proestrus, serum LH levels in control animals were 17.56 +/- 2.60 ng/mL, which were 920% above basal levels (1.90 +/- 0.13) indicating a spontaneous LH surge. This surge was suppressed in OH-F treated rats. Injection of LHRH, at the dose of 20 ng and above, reinstated the LH release in OH-F treated animals. Thus, the anti-androgen, OH-F, inhibits ovulation in PMSG-treated immature rats through its interference with the preovulatory LH surge; the inhibition can be reversed by hCG or LHRH. Hydroxyflutamide does not appear to interfere at the level of the pituitary, but may have direct action at the hypothalamic and (or) extrahypothalamic sites involved in the generation of positive feedback signals that control LH release. PMID- 3048619 TI - Autoregulation of blood flow in renal medulla of the rat: no role for angiotensin II. AB - Autoregulation of blood flow was assessed by a dual-slit technique in descending and ascending vasa recta of the exposed renal papillae of antidiuretic rats. There was complete autoregulation of blood flow in descending vasa recta. The lower limit of autoregulation was approximately 85 mmHg (1 mmHg = 133.3 Pa) and the upper limit was greater then 160 mmHg. Autoregulation in ascending vasa recta was also good. To test the role of angiotensin II in this autoregulation, the converting enzyme inhibitor captopril was infused. Captopril had no effect on autoregulation of blood flow in either descending or ascending vasa recta. We conclude that blood flow in vasa recta of renal medulla is efficiently autoregulated and that this autoregulation is independent of angiotensin II. PMID- 3048620 TI - Can blind infants and children use sonar sensory aids? PMID- 3048621 TI - Characteristics of verotoxigenic Escherichia coli from pigs. AB - Porcine verotoxigenic Escherichia coli were characterized with respect to frequency of occurrence, serogroup, and association with disease, weaning, and selected properties of the bacterium. Of 668 strains of E. coli from southern Ontario pigs with enteric disease, 32 (4.8%) produced verotoxin at 10(3)-10(7) cytotoxic doses per mL of culture supernatant. Of 22 isolates which belonged to O serogroups 138, 139 and 141, 15 produced verotoxin. Among other enterotoxigenic types of E. coli, two of 57 isolates of O157:K"V17" and two of 96 isolates of O149:K91 were verotoxigenic. The remaining 13 verotoxigenic E. coli belonged to O groups 2, 107, 120, 121 and 130. An additional 21 verotoxigenic E. coli belonging to O groups 138, 139 and 141 and three to O157:K"V17" were identified in a collection of 47 E. coli recovered from weaned pigs with enteric disease. Verotoxigenic E. coli were associated with postweaning diarrhea, bloody stools, sudden death and edema disease. They were isolated at similar frequencies (14%) from healthy weaned pigs, and from weaned pigs with enteric disease. Isolation rates from neonates were low and significantly different from rates in weaned pigs. Neutralizing antibody to verotoxin was not detected in the sera of 45 pigs, which included pigs from herds with a history of edema disease. Verotoxin was not associated with production of colicin, hemolysin, or enterotoxins or with any of 23 biochemical properties of the organisms. The serological data indicate that porcine verotoxigenic E. coli are not a common source of verotoxigenic E. coli for humans. Porcine verotoxin may play a role in postweaning diarrhea and absence of detectable neutralizing antibody in serum may be an important aspect of pathogenesis. PMID- 3048622 TI - Virulence differences among three strains of Haemophilus somnus following intratracheal inoculation of calves. AB - Pneumonia was induced in four month old Holstein calves by intratracheal inoculation of 1 x 10(9) colony forming units of Haemophilus somnus. Twenty calves were divided into four groups of five and challenged with a pneumonic strain (Group 1), an encephalitic strain (Group 2), a preputial strain (Group 3), or a placebo (Group 4). The clinical score, neutrophil count, respiratory rate, and temperature were significantly increased in group 1 by day 1 postinoculation (P less than 0.05) and maintained until day 6 postinoculation (P less than 0.05). The macroscopic pathological changes were significantly greater in group 1 (P less than 0.05). Haemophilus somnus was consistently isolated from pneumonic tissue of group 1 only. Groups 2 and 3 had mild transient increases in all parameters measured and macroscopically only small focal lesions were present. It is concluded that virulence differences exist between H. somnus strains following intratracheal challenge of bovine lungs. PMID- 3048624 TI - Traditional medicine of aboriginal Australia. PMID- 3048623 TI - Dietary aspects of adverse reactions to foods in adults. AB - Dietary considerations play an important role in the diagnosis, treatment and management of immunologic and nonimmunologic reactions to foods. Food diaries and trial elimination diets may prove helpful in identifying the responsible foods. Elimination diets must be monitored carefully for nutritional adequacy and should be used no longer than absolutely necessary; in some instances appropriate vitamin and mineral supplementation may be necessary. Ideally the identification of foods that provoke symptoms should be confirmed by means of double-blind challenge testing. Avoidance of some problem foods is unlikely to cause nutritional problems, but the practical and nutritional implications of allergies to staple foods such as cow's milk, eggs and wheat are far greater. Nonimmunologic adverse reactions that may mimic food allergic reactions include gastrointestinal disorders, sensitivity to food additives and psychologically based adverse reactions. There may be some degree of tolerance in metabolic disorders, which makes dietary management easier. Sensitivity to food additives necessitates careful scrutiny of food labels. In psychologic adverse reactions to foods, several foods are often involved, which increases the risk of nutritional problems. PMID- 3048628 TI - Clinical nurse specialization: an annotated bibliography--roles and functions. PMID- 3048626 TI - Skin tests. PMID- 3048625 TI - Eosinophils. Assays and interpretation. PMID- 3048629 TI - Hyperthermia and radiation therapy in the treatment of recurrent Merkel cell tumors. AB - A high incidence of local recurrence, spread to regional lymph nodes, and distant metastases has been reported after surgical excision of Merkel cell tumors (MCT). The use of postoperative radiation therapy and/or chemotherapy is reviewed from the literature. Despite adjuvant treatment, local tumor recurrences frequently develop. Two patients are presented with metastatic MCT recurrent in previously irradiated sites who had excellent clinical responses and local control following retreatment with local hyperthermia in conjunction with low to moderate dose radiation therapy. These patients represent the first reported use of hyperthermia in the management of MCT. The encouraging local responses described suggest a potential role for the use of hyperthermia and concomitant radiation therapy in the treatment of recurrent MCT. PMID- 3048630 TI - Pleuropulmonary blastoma. The so-called pulmonary blastoma of childhood. AB - The authors studied 11 pediatric intrathoracic neoplasms that share clinicopathologic features and constitute a specific tumor in children. These neoplasms were intrapulmonary, mediastinal, or pleural-based masses. A common histologic feature was the presence of small, primitive cells with blastematous qualities separated by an uncommitted stroma. Focal rhabdomyosarcomatous, chondrosarcomatous, and liposarcomatous differentiation was observed. Epithelial components had bland cytologic features and probably represented entrapped benign epithelium and/or mesothelium. The prognosis for these patients was grave; seven patients died of their disease 5 months to 2 years after diagnosis. Two patients have survived disease-free for 10 and 12 years after diagnosis. Two recent cases are alive 14 and 32 months after diagnosis. This neoplasm constitutes a distinct entity which has been reported in the literature as pulmonary blastoma in children. It differs from pulmonary blastoma in adults because of its variable anatomic location, primitive embryonic-like blastema and stroma, absence of a carcinomatous component, and potential for sarcomatous differentiation. The designation of pleuropulmonary blastoma is suggested by the authors for these intrathoracic neoplasms of childhood rather than pulmonary blastoma for histogenetic and anatomic reasons. The clinicopathologic features, immunophenotypic and ultrastructural characteristics, possible histogenesis, and differential diagnosis of these neoplasms from other thoracopulmonary tumors in children serve as the basis for this report. PMID- 3048631 TI - Flow cytometric analysis of pituitary tumors. Correlation of nuclear antigen p105 and DNA content with clinical behavior. AB - Flow cytometric quantitation of the proliferation-associated nuclear antigen p105 and DNA content was performed on nuclear suspensions from 12 paraffin-embedded pituitary macroadenomas and one pituitary carcinoma and correlated with clinical outcome. Median follow-up was 41 months (range, 33 to 48 months). Three of the 13 tumors (23%) had an identifiable aneuploid peak. Of the four tumors that recurred or metastasized, only one was aneuploid. Nuclear antigen analysis of all diploid tumors showed enhanced p105 expression in G2M phase cells compared to G0G1 cells. The G2M/G0G1 fluorescence ratio for p105 was consistently higher (P less than 0.05) for the three diploid tumors that recurred (median, 1.32 arbitrary fluorescence units; range, 1.27 to 1.80) than for the seven nonrecurrent diploid tumors (median, 1.20 arbitrary fluorescent units; range 1.14 to 1.22). These findings indicate a low incidence of DNA aneuploidy among pituitary tumors and suggest that for diploid adenomas, measurement of p105 may provide information useful in predicting prognosis and directing postoperative adjuvant therapy. PMID- 3048632 TI - Osteosarcoma of the uterine cervix associated with hyperplastic and atypical mesonephric rests. AB - The authors describe a unique case of osteosarcoma associated with atypical mesonephric rests occurring in the right lateral wall of the uterine cervix. The tumor was examined histologically, cytologically, immunohistochemically, and ultrastructurally and was compared with another osteosarcoma that filled an entire uterine cavity. Comparison with other neoplasms of the uterine cervix indicates that the lesion should be classified separately. Previously reported neoplasms associated with mesonephric rests within the cervix are also reviewed. PMID- 3048633 TI - Specific active lung cancer immunotherapy. Immune correlates of clinical responses and an update of immunotherapy trials evaluations. AB - The mechanisms of action of the specific active immunotherapy of solid tumors have not been defined. In an attempt to characterize some of these mechanisms, we report controlled studies of humoral immune responses and cell-mediated immune (CMI) responses in lung cancer patients with Stage I and Stage II adenocarcinoma and squamous cell cancer receiving pure tumor-associated antigen (TAA) specific active immunotherapy or combination immunochemotherapy. At 5 to 6 months postimmunotherapy, the humoral immune response measurements are predictive of response to therapy/survival in early lung cancer patients, permitting decisions as to whether to continue therapy. Patients with adenocarcinoma respond to combination chemoimmunotherapy by showing stronger or earlier responses to tests of immunity. Cell-mediated immunity to TAA at 17 to 24 months was far greater in patients receiving immunotherapy or immunochemotherapy compared with control patients, and also correlated with early humoral immune response and with 5-year survival. Here we report a further subset analysis of Stage I and Stage II lung cancer patients in a successful Phase III US specific active immunotherapy trial as substantiating the experience with Stage I patients in a successful Phase II Canadian trial. We analyze failures and suggest additional therapies, especially a chemoimmunotherapy trial indicated by our analyses of humorocellular immune variables reported here. PMID- 3048634 TI - Diet and cancer. Any progress in the interim? AB - It has been 5 years since the National Research Council (NRC) Committee on Diet, Nutrition and Cancer published "Interim Dietary Guidelines" for the nutritional prevention of cancer. The term "interim" implies that these recommendations should be regarded as temporary, pending more definitive findings from additional scientific research. This article reviews findings relevant to the connections between diet and cancer that have emerged from nutritional epidemiology subsequent to the 1982 the NRC report. Some recent research has supported the earlier work which served as a basis for the interim recommendations, some has not, and additional hypotheses have emerged. There continues to be evidence, although it is inconsistent, that dietary fat may be an important factor in colon cancer, and that something related to fruits and vegetables, perhaps carotene, may lower the risk of lung cancer. However, the hypothesized relationships between dietary fat and breast cancer and between dietary fiber and colon cancer have been less consistently supported by new findings. Meanwhile, a new hypothesis has emerged relating alcohol intake to breast cancer risk, although many important questions remain regarding the age at which alcohol use may affect risk, and the dose above which risk is increased. The last 5 years seem to have been characterized by only slow progress in our understanding of the relationship between diet and cancer. It is clear that in 1988 we are still very much in the interim. Critical methodologic assessments of the reasons for the discrepancies in findings among the various studies, and meta analytic approaches may be helpful in increasing our understanding of the set of epidemiologic research conducted to date. More important, however, are the many types of studies now underway, including more rigorously designed observational studies and chemo preventive and dietary-preventive trials. These studies will likely provide more definitive future answers to the questions we still face in the interim. PMID- 3048635 TI - Cancer prevention in the workplace and natural environment. A review of etiology, research design, and methods of risk reduction. AB - Guidelines for developing programs in cancer prevention and reduction of risk related to occupational and environmental carcinogens are provided in this review, which includes a summary of experimental and epidemiologic research designs for assessing potential etiologic agents, an overview of known occupational and environmental carcinogens, and a summary of general approaches to risk reduction. This review includes human cancer risks in the workplace and in the natural environment, which encompasses air pollution, water pollution, and hazardous waste disposal. Fourteen tables present the following: (1) established and probable carcinogens that may be encountered in the workplace, as contaminants of air or water, or as hazardous waste; (2) regulated carcinogens; (3) primary sources of exposure; (4) research designs; and (5) risk reduction methods. The interactive effects of multiple exposures are discussed as they relate to etiology and prevention. The importance of federal regulation is emphasized, as well as the need to understand the potential for cancer prevention in the context of broader economic, political and social issues. PMID- 3048627 TI - In vitro assays for immunoglobulin E. Methodology, indications, and interpretation. PMID- 3048636 TI - Electromagnetic radiations and cancer. Cause and prevention. AB - The various types of electromagnetic radiation differ considerably in their ability to induce cancer. The potential of radiofrequency or microwave radiation and low-frequency electromagnetic radiation to alter DNA is very limited, because their energy is too low to produce substantial ionizations. They are therefore unlikely to be carcinogenic by any direct mechanism. Epidemiologic studies of the carcinogenicity of microwave radiation are basically negative. Studies of workers with relatively high exposures to low-frequency electromagnetic fields have suggested that such persons may be at somewhat elevated risk for leukemia, especially of the acute myeloid type, but the studies have had methodologic weaknesses and mixed results. The association is not proven at this point, but neither can it be ruled out. For ionizing radiation, which is clearly carcinogenic, major questions pertain to how to define the magnitude of risk from low doses and low dose rates, how to identify subgroups of people who are especially susceptible to the effects of ionizing radiation, and how to minimize radiation exposure. When fortuitous radiation exposure from manmade sources, such as radioactive releases from nuclear power plants, are examined in the context of the total exposure people receive from natural sources, medical irradiation, etc., they are almost always found to be small by comparison. Quantitatively, two sources of radiation provide the greatest opportunities for exposure reduction: abatement of radon levels in homes, and reduction in medical radiation exposures. PMID- 3048637 TI - Steroid hormones and medications that alter cancer risks. AB - Alkylating agents have caused acute nonlymphocytic leukemia (ANLL), probably bladder cancer, and possibly other solid tumors. Phenacetin also has enhanced risk of bladder cancer, and probably also carcinoma of the renal pelvis. Topical nitrogen mustard, potassium arsenite, tar ointments, and methoxsalene have been related to development of nonmelanotic skin cancers. Immunosuppression by azathioprine, usually with prednisone, has enhanced risks of non-Hodgkin's lymphomas, hepatobiliary cancers, and various mesenchymal tumors. Liver cancers have been reported in users of androgenic anabolic steroids, and both hepatic cell adenomas and carcinomas have been associated with use of combined oral contraceptives. These contraceptives reduce risks of endometrial and ovarian carcinomas. Estrogens increase risk of endometrial cancer. Exposure to diethylstilbestrol in utero can result in clear cell carcinomas of the vagina and cervix, and possibly testicular carcinomas. PMID- 3048638 TI - Cancer screening. Degrees of proof and practical application. AB - The purpose of this paper is to clarify the short-term and long-term objectives of screening for various cancers, and to indicate the kinds of data that are needed to determine whether or not the objectives are met. Cancers at various sites differ with respect to their innate suitability for screening. Criteria that enhance screening suitability include the potential for serious complications and a high rate of mortality (applicable to most cancers), a prolonged preclinical phase, and an existing therapy that is simpler and more effective in reducing the mortality rate when applied to preclinical disease than to clinically evident cancer. Tests and procedures suitable for screening are simple to perform, inexpensive, acceptable to patients and physicians, safe, relatively painless, and accurate, as measured by the test's sensitivity and specificity. The actual yield of previously undiagnosed cancer arising from a screening program will depend heavily on prevalence of disease in the screened population, specificity of the screening test, and successful follow-up of screen positive patients with diagnosis and treatment. These issues are discussed in the context of four cancers and their respective screening modalities: cervical cancer and cytologic studies, breast cancer and mammography, colon cancer and fecal occult blood tests, and lung cancer and sputum cytologic studies. The quality of data on which screening decisions have been made for each of these cancers and tests varies. The cancers vary in terms of their relevant biologic characteristics and treatment effectiveness. Similarly, each screening procedure has its own particular advantages and disadvantages. Current American Cancer Society Guidelines for early detection of three of the cancers are presented. PMID- 3048639 TI - Urologic cancer. AB - Cancer prevention and risk reduction in patients with carcinomas of the urinary tract are most easily identified by smoking cessation. Recent assessments of patients with bladder carcinomas show a high percentage of active smokers. Urologic surgeons and those dealing with cancers of the genitourinary tract should recognize such an important factor. In other lifestyle guidelines, the American Cancer Society suggests that those who significantly reduce the fat content of their diet with weight reduction will generally reduce risk factors in a number of cancers, including prostate cancer. Many new diagnostic tests have been introduced since the 1980 conference. Transrectal ultrasonography as a preoperative staging technique has been received with some enthusiasm. Currently, a national program for the testing of the detection of prostate cancer by ultrasound compared with other physical examinations and marker antigens, such as prostate antigen, is underway. The evaluation of renal masses continues to be improved by the use of ultrasonography and computed tomography (CT) scanning. Currently, magnetic resonance imaging (MRI) techniques are believed to have added some improved assessment for particularly large tumors and the question of their vascular extension. Occasionally observed lesions of the adrenal gland without hormonal expression in patients continue to be noticed during evaluations for frequently unrelated diagnoses. The management of such lesions less than 3.5 cm in diameter is believed to be conservative based on their size. Preoperative staging of the bladder has improved with transurethral ultrasonography use and the follow-up with patients has developed more frequently with the widely available urinary cytology. Fine needle aspiration of the prostate gland, lymph nodes, and other urinary tumor masses is increasing in diagnostic use. Flow cytometry has been applied to all lesions of the urinary tract and continues to be a field of clinical investigation. The field of biologic markers in diagnosis, detection, and follow-up evaluation of patients with carcinomas of the urinary tract is a most interesting and progressive one. Currently, the most widely accepted new innovation is the prostate antigen, which was developed by members of the National Prostatic Cancer Project between 1975 and 1986. Other markers offer potential added value in more accurate and earlier diagnosis and detection and correlation with prognostic factors concerning individual groups of patients. Progress in the detection, diagnosis, and management of genitourinary cancer is being achieved, and prevention, although perhaps simplistic in design, can be more readily applied. PMID- 3048640 TI - The National Cancer Institute's intervention trials. AB - The National Cancer Institute (NCI) has increased its emphasis in cancer prevention by undertaking a number of human intervention trials. Special emphasis is placed on chemoprevention, diet, and smoking and tobacco use. Some 24 clinical trials are ongoing in chemoprevention to assess the role of specific chemicals (natural and synthetic) in preventing, inhibiting, or reversing carcinogenesis. In diet and nutrition, several macronutrient trials are underway. The most significant of these is examining the relationship between dietary fat and breast cancer. In 1982, the NCI initiated a broad intervention research effort in smoking and tobacco use which has resulted in 46 prevention and clinical trials covering 25 states and more than 200 cities. More recently, a North American community-based intervention trial was begun to test cessation strategies for heavy smokers in 22 different sites. All of these intervention research efforts are testing strategies which later can be applied to large target population thereby supporting NCI's ambitious goal to reduce cancer mortality in this nation 50% by the year 2000. PMID- 3048641 TI - Advances in imaging. AB - The introduction of sonography, x-ray computed tomography, magnetic resonance imaging (MRI), and magnetic resonance spectroscopy (MRS) have enhanced the radiologist's ability to delineate and stage neoplasms in all parts of the human body. Images of excellent quality can be generated within a reasonable time frame and with minimal biologic risk. All of the more sophisticated imaging modalities are costly and none can, isolated from clinical data, provide histologic diagnoses. Within the next few years it is anticipated that the speed of magnetic resonance image acquisition will increase and that contrast agents for MRI will be brought into clinical use. Evaluation of cost-effectiveness will continue and new diagnostic algorithms can be expected to evolve. PMID- 3048642 TI - Emotions and cancer. New perspectives on an old question. AB - This article outlines current issues in the study of psychological factors and cancer: (1) the role of psychological factors in disease promotion and progression, (2) the impact of neoplastic disease and its treatment on psychosocial functioning, and (3) the long-term impact of cancer and cancer treatment on the survivor and issues of family bereavement following the death of a cancer patient. The discussion ends with a consideration of future directions for research. PMID- 3048643 TI - Sequential trial of initial chemotherapy for advanced cancer of the head and neck. DDP versus DDP + 5-fluorouracil. AB - The study, which compares DDP to DDP + FU, was planned to detect an increase by 60% in efficacy and by 5% in toxicity (2a = B = 5%) for DDP + FU. In a previous trial DDP produced 15% of responders and 5% of high-level toxic manifestations. The eligible patients with an advanced head and neck cancer were paired off successively on the basis of the tumour site and the UICC stage. DDP (100 mg/m2; day 1) was administered with hyperhydration, alone in the first protocol and followed by a 5-day continuous infusion of 5-FU (1 g/m2) in the second one. Courses were repeated every three weeks. Assessment was carried out after three courses or two, in cases of toxic manifestations. Seventy-four patients, who were paired off, entered the trial. The median age was 55 years and the median Karnofsky index was 90. The tumor site was as follows: 28 hypopharynx, 28 oropharynx, 8 oral cavity, and 10 multiple primary cancers. According to the UICC stage, there were 14 T1/T2 N3, 60 T3/T4 with among them 45 N3, and they were all MO. Comparisons were made through sequential closed plans. The combination chemotherapy was more efficacious than DDP with a difference that could be appreciated by the sequential analysis as high as 60% (95% confidence interval, 38% to 82%). The high-level toxicity appeared more significant (+25%) for the association. After radiation therapy 11 of 37 patients (30%) achieved a complete response in the arm with DDP versus 18 of 37 (49%) in that with DDP + FU. The median survival times were 9 and 11.5 months, respectively, and were not statistically different. PMID- 3048644 TI - Adjuvant chemotherapy in clinical stage M0 sarcoma of soft tissue. AB - The clinical course of 117 patients who were treated for Stages IIB-IIIC sarcoma of soft tissues by the combined approach of radiation and surgery at MGH during the period 1971-1984 have been analyzed for an effect of adjuvant chemotherapy to improve clinical results. Thirty-two patients received adjuvant chemotherapy and 85 were treated by local methods alone. The chemotherapy protocols featured doxorubicin (25 patients) or VAC (seven patients). The no chemotherapy patients were controls matched for tumor grade and size and were treated over approximately the same period. The two groups were not well matched for age: median age of 38.5 and 54 years for the treated and control groups, respectively. There was no significant difference in survival. However, the time to first metastasis was longer in the treated group, P = 0.04. Furthermore, the local failure rate appeared to be lower in the treated group, P = 0.06. PMID- 3048645 TI - Primary rhabdomyosarcoma of the kidney. A light microscopic, immunohistochemical, and electron microscopic study. AB - Primary rhabdomyosarcoma of the kidney is a rare and highly aggressive tumor in the adult population. A case is reported in a 70-year-old woman with the diagnosis confirmed by immunohistochemistry and electron microscopy. This is the first case studied using the immunoperoxidase technique and the second with electron microscopic examination. To make a diagnosis of primary sarcoma, of the kidney, three criteria must be met: (1) a metastatic sarcoma must be ruled out; (2) the tumor must arise from renal parenchyma; and (3) a sarcomatoid variant of renal cell carcinoma needs to be excluded. The literature is reviewed and available clinical and pathologic details are summarized. PMID- 3048646 TI - Breast cancer epidemiology. PMID- 3048647 TI - Characterization of drug metabolism enzymes in estrogen-induced kidney tumors in male Syrian hamsters. AB - In an attempt to characterize metabolism enzymes of the estrogen-induced kidney tumor in male Syrian hamsters, the activities of enzymes involved in drug and glutathione metabolism were determined in tumor tissue. Kidney tumors were induced in male Syrian hamsters by treatment with estradiol for 8 months. Cytochrome P-450 and cytochrome b5 concentrations in tumors were below detectable levels. However, when cytochrome P-450-mediated oxidation was analyzed by product formation assays, the oxidation of E-diethylstilbestrol to diethylstilbestrol 4',4"-quinone by tumor microsomes was 10-20% of the rate found in control microsomes. In kidney tissue surrounding estrogen-induced tumors, cytochrome P 450 and b5 contents were 50-60% less than those in untreated kidney. Activities of reducing enzymes of drug metabolism (cytochrome P-450, cytochrome b5 and NADH:cytochrome c reductases), glutathione metabolism enzymes (glutathione peroxidase, glutathione transferase, glutathione reductase, and gamma-glutamyl transpeptidase), and free radical scavenging enzymes (superoxide dismutase, catalase, and quinone reductase) in tumor were significantly lower than in untreated kidney tissue. The activities of these enzymes in renal tumor surrounding tissue were between those observed in tumor and control kidney. Glucose-6-phosphate dehydrogenase activity was increased by 50% in surrounding tissue and 430% in tumor compared to values in untreated controls. The decreased enzyme activity levels in hormone-exposed tissue surrounding tumors likely represented an adaptation of this tissue to the neoplastic environment induced by chronic estrogen treatment. PMID- 3048648 TI - Incidence and possible clinical significance of K-ras oncogene activation in adenocarcinoma of the human lung. AB - 47 tumor samples, 45 of which were obtained at thoracotomy for non-small cell lung cancer were examined for mutational activation of the oncogenes H-ras, K ras, and N-ras. A novel, highly sensitive assay based on oligonucleotide hybridization following an in vitro amplification step was employed. ras gene mutations were present in nine of 35 adenocarcinomas of the lung (all K-ras), in two of two lung metastases of colorectal adenocarcinomas (1 x K-ras, 1 x N-ras) and in one adenocarcinoma sample obtained at autopsy (H-ras). All K-ras and H-ras mutations were in either position 1 or 2 of codon 12, while the N-ras mutation was in position 2 of codon 61. The potential clinical significance of K-ras activation was analyzed using the combined results of this and of our earlier study (S. Rodenhuis et al., New Engl. J. Med., 317: 929-935, 1987). Lung adenocarcinomas with K-ras mutations tended to be smaller and were less likely to have spread to regional lymph nodes at presentation. With a median follow up of 10 months, survival data are still immature. None of six adenocarcinomas of nonsmokers had a K-ras mutation and only one of four who had stopped smoking more than 5 years before. We conclude that mutational K-ras activation is present in about a third of adenocarcinomas of the lung and that the mutational event may be a direct result of one or more carcinogenic ingredients of tobacco smoke. Studies involving larger numbers of patients are required to confirm the association of K ras activation with smoking and the inverse relation with tumor progression. PMID- 3048650 TI - A novel method for the detection of necrotic lesions in human cancers. AB - Data are presented in support of the hypothesis that malignant tumors, containing abnormally permeable, degenerating cells, can be selectively detected using monoclonal antibodies to intracellular antigens. Biodistribution, imaging, and autoradiographic studies were performed in nude mice transplanted with four different human tumor cell lines to demonstrate the binding of radiolabeled antinuclear monoclonal antibodies within bulky tumors containing necrotic lesions. For these studies, two monoclonal antibodies, designated TNT-1 (IgG2a) and TNT-2 (IgM) were chosen since they were found to bind to abundant nuclear antigens which are retained in permeable, dying cells. F(ab')2 fragments prepared by pepsin digestion were radiolabeled with iodine-125 or iodine-131 by the iodogen method for i.v. administration. Biodistribution studies in nontumor bearing BALB/c mice at various time intervals revealed normal patterns of antibody excretion with no accumulation of antibody in healthy organs. In contrast, biodistribution studies performed on Day 3 in tumor-bearing nude mice showed high tumor to organ ratios in those animals bearing necrotic tumors. Necrotic regions dissected at necropsy gave tumor to blood ratios as high as 131:1. Transplants having little demonstrable necrosis were found to have low tumor to blood ratios (0.4:1). Sequential imaging studies confirmed the high tumor-to-organ ratios and showed positive tumor imaging as early as 4 h. Autoradiographic studies of excised tumors showed the presence of label selectively in necrotic areas with preferential labeling over the nuclei of degenerating cells. Because of the universal presence of these nuclear antigens and the known prevalence of necrosis in tumors, this approach may be of value for the imaging and treatment of a wide variety of cancers in humans. PMID- 3048649 TI - Effect of retinoic acid on phorbol ester-stimulated differentiation and protein kinase C-dependent phosphorylation in the U937 human monoblastoid cell. AB - Phorbol esters stimulate differentiation of certain human leukemic cell lines. Although activation of protein kinase C may mediate certain effects of phorbol esters, controversy exists as to the role of protein kinase C activation in phorbol ester-induced differentiation. Retinoic acid modulates responses to phorbol esters in several cell types. Retinoic acid has also been found to alter protein kinase C-dependent phosphorylation in leukemic cells. We correlated the effects of retinoic acid on protein kinase C-dependent phosphorylation and differentiation stimulated by 12-O-tetradecanoylphorbol-13-acetate (TPA), a phorbol ester, in the human monoblastoid U937 cell line. At concentrations less than 1 nM, which were 100-fold less than those directly stimulating differentiation, retinoic acid potentiated TPA-induced differentiation of the U937 cell as assessed by enhanced adherence to plastic and acquisition of nonspecific esterase activity. TPA-stimulated decreases in cellular proliferation were not affected by retinoic acid treatment. Without altering the sensitivity to TPA, retinoic acid increased the maximal response to this agent. Retinoic acid enhanced TPA-stimulated phosphorylation of a Mr 48,000 substrate in intact 32P labeled U937 cells and also increased the protein kinase C-dependent phosphorylation of a similar Mr 48,000 substrate and a Mr 80,000 substrate in cellular extracts. In cellular extracts the retinoic acid-induced enhancement of protein kinase C-dependent phosphorylation was predominantly localized to the cytosolic fraction. Increases in protein kinase C-dependent phosphorylation were evident within a 12-h exposure to 1 nM retinoic acid and were observed at retinoic concentrations of 0.01 to 1 nM. A retinoic acid-induced increase in the protein kinase C-dependent phosphorylation of an exogenous substrate, histone, was observed following diethylaminoethyl extraction of cytosol, but not a solubilized particulate fraction. The conditions of retinoic acid treatment increasing protein kinase C activity and enhancing protein kinase C-dependent phosphorylation of endogenous substrates were similar to those conditions potentiating phorbol ester-induced differentiation. Thus, the retinoic acid induced amplification of phorbol ester signal transduction at the level of protein kinase C activation could mediate the effects of this vitamin on phorbol ester-induced differentiation. PMID- 3048651 TI - Growth requirements for human acute lymphoblastic leukemia cells: refinement of a clonogenic assay. AB - Since freshly obtained acute lymphoblastic leukemia (ALL) cells rarely replicate spontaneously in vitro in a sustained way, development of a useful clonogenic assay for ALL blast progenitors is dependent on identifying the cellular growth requirements. Thus, marrows from 25 ALL cases were cultured in methylcellulose to determine the optimal conditions for cell growth. Blast colonies were confirmed as leukemic by morphology, cytochemistry, surface markers, and cytogenetics. Irradiated (7000 rads) normal peripheral blood feeder cells were an absolute requirement and produced number-dependent increases in ALL colonies; added growth factors enhanced the feeder cell effect. ALL cell-feeder cell contact was essential since their physical separation in a two-layer culture system drastically interfered with colony growth. Feeder cells from various donors, including new and relapsed cases of ALL, yielded colony numbers that differed widely when tested on the same marrow with and without added growth factor; thus, identification of a "good" feeder cell donor was key to an optimal assay. Neither recombinant interleukin-2 nor recombinant GM-CSF had ALL growth-promoting properties when tested alone or in combination but in the presence of feeder cells they moderately enhanced the feeder cell effect. The most effective growth factors were derived from cells exposed to phytohemagglutinin (PHA) for 72 h. In order of magnitude for colony growth-promoting activity, PHA-T cell conditioned medium (CM) was more stimulatory than PHA-blast cell CM followed by PHA-leukocyte CM; removal of PHA from CM by affinity chromotography did not alter the results. The most potent PHA-TCM was prepared from T-cells from a phlebotomized hemochromatosis patient; PHA-TCM from transfused thalassemia patients and normal donors were less active. Concanavalin-A blast cell CM had modest colony promoting properties whereas CM prepared with other B-cell mitogens and supernatants from ALL blasts in liquid culture had none. Our studies illustrate the complex and fastidious growth needs of ALL cells. The data have allowed us to refine a clonogenic blast progenitor assay that should facilitate study of proliferative properties of B and T lineage leukemias. The assay could be adapted further for detection of residual leukemia cells in marrow samples used for autologous transplantation, and in patients during complete hematological "remission." PMID- 3048652 TI - Genetic convergence and divergence in tumor progression. PMID- 3048653 TI - Early cell motility changes associated with an increase in metastatic ability in rat prostatic cancer cells transfected with the v-Harvey-ras oncogene. AB - The development of metastatic ability by cancer cells is a multifactorial process whose temporal events are complex and poorly understood. One step in the metastatic process may involve cell motility. Previous studies reported correlations between motility and metastatic ability. Whether this correlation, seen in cancer cells maintained for long periods of time, is an epiphenomenon developing late in the growth of the cancer as a selection artifact of continuous passage, or is critically required for the acquisition of metastatic ability is unknown. To investigate the relationship between cell motility and the acquisition of metastatic ability, advantage was taken of recently developed DNA transfection methods for inducing high metastatic ability in initially low metastatic cancer cells. The Dunning AT2.1 cell line, a clonal rat prostatic cancer cell line with low metastatic ability, was transfected with a plasmid containing the neomycin resistance gene alone or in combination with the v-Harvey ras oncogene. A series of the transfected cells was isolated by limiting dilution. After the first in vitro passage following transfection, cells were inoculated into rats to characterize their metastatic ability. The same transfectants were simultaneously studied using our visual grading system of cell motility to study the early motility changes associated with newly acquired metastatic ability. The data demonstrate increased membrane ruffling, pseudopodal extension, and cell translation (translocation) in the v-H-ras-transfected cell lines with high metastatic potential. PMID- 3048655 TI - Modulation of HT-29 human colonic cancer cell differentiation with calmidazolium and 12-O-tetradecanoylphorbol-13-acetate. AB - The effects of a protein kinase C activator, 12-O-tetradecanoylphorbol-13-acetate (TPA), and of a calmodulin antagonist calmidazolium (CZ), on a human colonic cancer cell line HT-29 were analyzed. HT-29 cells are undifferentiated in standard culture conditions (HT-29 G+) and display an enterocytic differentiation when cultured in glucose-deprived medium (HT-29 G-). Early effects of TPA and CZ on the localization of cytoskeletal proteins (caldesmon, alpha-actinin and vinculin) and on cell proliferation were examined. Differentiation of the cells was assessed after 4 weeks on the basis of ultrastructural and functional characteristics of enterocytic polarity, presence of apical brush borders, expression of brush border membrane antigens (Caco 5/50 and sucrase-isomaltase), and segregation of calmodulin to the brush border cytoskeleton. TPA treatment of HT-29 G+ or G- cells induced early morphological and cytoskeletal alterations: the cells rounded up and lost their stress fibers with the associated caldesmon, alpha-actinin, and vinculin. TPA did not modify the differentiation of G- cells, but induced in G+ cells the expression, although limited, of enterocytic differentiation characteristics. Addition of CZ to HT-29 G- cells enhanced their differentiation state but did not provoke any early morphological or cytoskeletal alterations. No effects of CZ on HT-29 G+ cells were obvious. The results suggest that protein kinase C, the TPA receptor, is involved in the triggering of HT-29 G+ cell differentiation whereas calmodulin-dependent functions would be implicated in HT-29 G- cell maturation. PMID- 3048654 TI - In vivo effects of recombinant human interleukin 2 on antitumor and antiviral natural immunity in induced or natural murine immunodeficiency states. AB - We have examined the ability of in vivo treatment of mice with recombinant interleukin 2 (rIL-2) to affect natural immunity measured against tumor (YAC-1) or virally infected (herpes simplex type 1) target cells. rIL-2 treatment leads to significant increases in natural killer/lymphocyte-activated killer (NK/LAK) function and spleen cells recovered. This effect is dose dependent and strain related. The latter parameter correlated with the pretreatment NK activity level of the strain. The rIL-2 induced NK/LAK augmentation is also kinetically restricted as treatment must have occurred within 48-72 h of assay to be effective. The rIL-2 therapy effectively enhances both antitumor and antiviral NK/LAK activity and results in a noticeable increase in asialo-GM1-positive cells in the spleens of treated mice as well as a significant increase in IL-2 receptor expression as monitored by either cytometry or radioligand binding. In vivo treatment of mice with an antibody directed to the ASGM1 determinant effectively reduces the rIL-2 augmentation of both antitumor and antiviral activity even though this treatment does not affect the pretreatment level of antiviral activity. Various natural and induced immunodeficiency states (immunotherapy, irradiation, immunosuppressive drugs, cytoreductive drugs) have been examined for the ability of in vivo treatment with rIL-2 to enhance NK/LAK activity. In vivo rIL-2 administration is differentially effective in enhancing NK/LAK activity in these situations. Notably, in these induced immunodeficiency states, although NK/LAK activity is commonly enhanced, the number of spleen cells recovered often is only marginally affected. Thus, as expected, a limiting aspect in this use of a natural immunomodulator is the number of potentially responsive cells present in the immunodeficiency condition. In addition, correlations between rIL-2 effect, several of the immunodeficiency states, and vascular leak syndrome are briefly discussed. PMID- 3048656 TI - Development of a dual label fluorescence technique that can be utilized to elucidate the mechanism of action of monoclonal antibody-drug conjugates. AB - A method is described that allows the simultaneous visualization and relative assessment of both the antibody and drug components of monoclonal antibody-drug conjugates at the target cell membrane. The antibody is detected by a fluorescein conjugated anti-mouse immunoglobulin serum while the drug is visualized by rhodamine avidin or phycoerythrin-streptavidin binding to a biotinylated anti Vinca alkaloid monoclonal antibody. This technique was effective in demonstrating the cell surface localization of a monoclonal antibody-Vinca alkaloid conjugate to human lung adenocarcinoma cells grown in vitro and was also used to demonstrate targeting of the conjugate in vivo to the membranes of these same tumor cells grown as a nude mouse xenograft. This method was also utilized to help elucidate the mechanism of action of monoclonal antibody-drug conjugates. PMID- 3048660 TI - Structure of a neutral polymer isolated from the lipopolysaccharide of the reference strain (C.D.C. 4523-60) for Serratia marcescens serogroup O15. PMID- 3048658 TI - Intracranial extracerebral glioneural heterotopia. AB - A newborn baby girl with intracranial extracerebral glioneural (brain) heterotopia in the right frontoparietal area is described. The heterotopic brain was predominantly composed of neuronal and glial elements, with partial cerebellar differentiation. It also contained well-differentiated adipose tissue. The possible mechanism for its origin and the causes of central nervous system heterotopia are discussed. PMID- 3048657 TI - Moyamoya disease. AB - The authors reviewed the Japanese literature on moyamoya disease. In the article we discuss the history of such investigations in Japan, the signs and symptoms, the diagnosis (especially concerning diagnostic criteria and magnetic resonance imaging), the pathology in relation to its etiology, and the current methods of treatment. On the whole, the main aim of the paper was to introduce our concept of moyamoya disease that is now current in Japan. PMID- 3048659 TI - Structure of the capsular polysaccharide (K19 antigen) from uropathogenic Escherichia coli O25:K19:H12. PMID- 3048661 TI - [Isosorbide dinitrate spray: a comparative pharmacodynamic study with a sublingual preparation in patients with chronic cardiac insufficiency]. PMID- 3048663 TI - [The artificial heart: state of the art and new prospectives]. PMID- 3048662 TI - [Contribution of digital angiography to the evaluation of ventricular and coronary imaging]. PMID- 3048664 TI - [Iatrogenic arrhythmia]. PMID- 3048665 TI - [Outcome of the normotensive subject in a 5-to-10 year follow-up: normal arterial pressure or hypertension?]. PMID- 3048666 TI - [Transient electrocardiographic aspect of left bundle-branch block with right axial deviation appearing during lateral subendocardial infarct]. PMID- 3048667 TI - Recurrent ventricular tachycardia in hypothyroidism--report of a case and review of the literature. AB - A 47-year-old woman suffered from recurrent attacks of ventricular tachycardia. Her electrocardiogram showed low voltage, right bundle branch block and prolonged QT interval. Hormonal studies disclosed primary hypothyroidism. The arrhythmia responded to treatment with procainamide and did not recur following thyroid replacement therapy. The QT interval returned to normal. Five similar cases reported in the literature are reviewed, emphasizing the importance of QT prolongation, in the context of hypothyroidism, as a risk factor for the occurrence of ventricular tachycardia. PMID- 3048668 TI - Assessing cardiac function by gas exchange. AB - Exercise stresses the primary function of the cardiovascular system, which is the supply of O2 and removal of CO2 from the cells of the body. Even ordinary walking requires an increase in O2 consumption and CO2 production by the exercising muscles of 20 times the resting level. While pulmonary dysfunction may affect arterial blood gas tensions, the dynamics of O2 uptake and CO2 output by the lungs depend on the circulatory responses to exercise. Thus, measurement of the dynamics of O2 uptake in response to exercise has been shown to reflect cardiovascular function. Inability of the circulatory responses to meet an increased O2 requirement may be reflected in abnormalities in O2 uptake dynamics, and an early increase in CO2 output relative to O2 uptake consequent to bicarbonate buffering of lactic acid. Application of currently available technology for the continuous measurement and analysis of pulmonary gas exchange can afford the practicing or investigative cardiologist with a noninvasive and inexpensive means for assessing cardiovascular function. PMID- 3048670 TI - The pathology of acute myocardial infarction: definition, location, pathogenesis, effects of reperfusion, complications, and sequelae. AB - This article reviews the definition, locations, and pathogenesis of acute myocardial infarction and provides a review of the pathology of acute reperfusion therapy. Various complications and sequelae of acute myocardial infarction are also surveyed. PMID- 3048669 TI - Psychosocial and physical rehabilitation after heart transplantation: 1-year follow-up. AB - Experience on the rehabilitation of 62 heart-transplanted patients with a mean follow-up period of 15 months and a total survival rate of 79% is reported. From the present study we may conclude that: (a) One month after surgery, oxygen consumption of transplanted patients compared to coronary artery bypass-grafted patients was statistically lower (p less than 0.025). An excess ventilation was observed in transplanted patients in relation mainly to an excessive increase in blood lactates. (b) Improvement of maximal working capacity observed immediately after grafting was still enhanced after 1 year of a comprehensive rehabilitation program (p less than 0.001). This improvement was more related with an improvement of the respiratory function and of the peripheral factors than with a circulatory effect. (c) Four months after transplantation 71% of the patients still at work 6 months before operation returned to work. (d) The quality of life, well-being and heart acceptation demonstrated an immediate increase in physical items after transplantation while psychosocial items decreased postoperatively and normalized after weeks or months. PMID- 3048671 TI - The role of anticoagulation in acute myocardial infarction. AB - The rule of anticoagulation therapy in the setting of an acute myocardial infarction has been debated for decades. The role of such therapy in reducing mortality, preventing deep venous thrombosis and pulmonary emboli, and in reducing the frequency of left ventricular thrombus formation and subsequent systemic embolization is discussed. PMID- 3048672 TI - Management of post-infarction angina. AB - Post-infarction angina includes a syndrome of ischemic chest pain occurring either at rest or during minimal activity 24 hours or more following an acute MI. It develops in approximately 10 to 15 per cent of patients and is particularly common in non Q-wave infarcts involving the anterior myocardial wall. Post infarction angina may result from ischemia either within the infarct zone or at a distance and frequently portends a poor long term prognosis. Platelet aggregation, coronary vasospasm, and thrombus formation at the site of a ruptured atherosclerotic plaque are each involved in its pathogenesis. The initial treatment of post-infarction angina includes identification and correction of factors that increase myocardial demand including congestive heart failure and arrhythmias. beta-Adrenergic blockers, calcium channel blockers, and nitrate preparations constitute the first line of medical therapy. The role of heparin is controversial, yet it continues to be used in clinical practice. Although thrombolytic agents are currently being investigated, early experience suggests that they may accelerate clinical stabilization and allow time for elective revascularization when required. Antiplatelet therapy with aspirin has proven benefit in the long term management of unstable angina. Patients unresponsive to medical therapy should be considered for intra-aortic balloon pump placement and early coronary angiography. Revascularization with either coronary angioplasty or coronary artery bypass grafting may then be performed as dictated by the overall clinical status, available facilities, and coronary anatomy. PMID- 3048673 TI - Ventricular arrhythmias: medical therapy, device treatment, and indications for electrophysiologic study. AB - Sustained ventricular arrhythmias occur most frequently during the first several months following MI, although the risk of arrhythmia development continues for many years. The mechanism responsible for most of these arrhythmias is a re entrant circuit located in the border zone between the normal and scarred subendocardium. Clinical factors that identify patients at greatest risk for developing sustained ventricular arrhythmias after MI include depressed left ventricular function, acute left ventricular aneurysm formation, electrical instability, new bundle branch blocks, and residual ischemia. Attempts to lower the arrhythmic risk of these patients is a major area of clinical investigation. Although frequent asymptomatic ventricular ectopy and nonsustained ventricular tachycardia are risk factors for sudden death after infarction, it has not been demonstrated that empiric treatment of these arrhythmias with antiarrhythmic agents improves survival. Electrophysiologic studies have significantly contributed to understanding the mechanisms responsible for sustained ventricular arrhythmias. Although the role of electrophysiologic studies in guiding therapy in patients with sustained ventricular tachycardia or sudden death after infarction has been well established, their utility to identify high risk subgroups after infarction has not been conclusively determined. New treatment modalities have resulted in an improved outcome in patients with malignant ventricular arrhythmias (Table 2). These strategies include new pharmacologic therapies, arrhythmia surgery, use of automatic implantable cardioverter defibrillators or antitachycardia pacemakers, and percutaneous catheter ablation of the re-entrant circuit responsible for these arrhythmias. PMID- 3048674 TI - Bradyarrhythmias, abnormalities of conduction, and indications for pacing in acute myocardial infarction. AB - Bradyarrhythmias and conduction disturbances are not infrequently observed in association with acute MI. The sinus node artery is supplied by the right coronary circulation only slightly more often than the left. As a result of concomitant vagotonia, however, sinus node dysfunction is more common with inferior infarction. This influence, as well as a predominantly right-sided circulation, also makes AV nodal block more frequent with such infarctions. Bradyarrhythmias due to sinus or AV nodal dysfunction often require only observation. If symptomatic, they are usually responsive to vagolytic or chronotropic drugs, but may necessitate pacemaker therapy often only on a temporary basis. The distal conduction system including the bundle branches is supplied mainly, but not exclusively, by the left anterior descending artery. Thus, acute bundle branch block is often associated with anterior MI. The indications for both temporary and permanent prophylactic pacing in this situation remain controversial. Several authors have made recommendations based on risk stratification. We would temporarily pace patients with anterior or indeterminate infarctions and new right or left bundle branch block, and probably those with bilateral bundle branch block of indeterminate age. All patients with new bilateral or alternating bundle branch block should be paced, regardless of infarct site. Permanent prophylactic pacing would appear indicated in patients exhibiting alternating bundle branch block or perhaps new right bundle branch block and left posterior hemiblock. In contrast to this group, the treatment of patients who develop sudden complete heart block, whether transient or permanent, is clear-cut. These patients require continuous (temporary followed without interruption by permanent) pacemaker therapy (Table 3). PMID- 3048675 TI - Treatment of the hemodynamically unstable patient. AB - The pathogenesis, diagnosis, and management of six clinical syndromes associated with acute myocardial infarction and hemodynamic instability are discussed: (1) autonomic disturbances with hypertension-tachycardia or hypotension-bradycardia; (2) pulmonary edema; (3) cardiogenic shock; (4) right ventricular infarction; (5) rupture of ventricular free wall or septum; and (6) papillary muscle rupture. PMID- 3048676 TI - The role of nuclear cardiology in assessment of acute myocardial infarction. AB - In summary, nuclear techniques have important roles in both the acute and convalescent phase of MI. Acutely, both myocardial perfusion and ventricular function may be assessed. In patients treated with thrombolytic therapy, perfusion imaging, and metabolic imaging using PET may be able to delineate ischemic myocardium. These techniques are being developed to accurately quantify salvaged myocardium for assessing the efficacy of an acute intervention, and to characterize (functionally) the remaining myocardium for possible further intervention. In the convalescent phase, prognostic information is obtained from both rest and exercise evaluation of ventricular function and reserve, ambulatory monitoring, as well as from the presence of exercise-induced ischemia as documented by quantitative thallium imaging. The results of these studies can be used to determine appropriate management strategies. PMID- 3048677 TI - An update on cardiac enzymes. AB - Determination of plasma total and MB CK concentration provides accuracy superior to any other currently available method for the diagnosis of acute MI. Nevertheless, elevation of MB CK is occasionally detected in the absence of acute MI for several reasons, some of them assay-dependent; consequently, the clinician should suspect a noncardiac source of MB CK or a spurious result if assay determinations are discordant with the clinical setting. In addition to providing precise diagnosis of acute MI, quantitative MB CK assays can also be used to obtain an accurate estimate of infarct size. In recent years, accuracy in the diagnosis of acute MI has assumed even greater importance, since the choice and timing of a variety of diagnostic and therapeutic options following coronary care unit admission hinge on whether infarction has occurred. Furthermore, the advent of thrombolytic therapy of acute MI has emphasized the need for more sensitive biochemical markers of necrosis in the first hours; the newly discovered subforms of MM and MB CK show promise as a means of proving an early diagnosis of acute MI and also as a means of assessing reperfusion noninvasively. PMID- 3048678 TI - Pathophysiology and differential diagnosis of restrictive cardiomyopathy. PMID- 3048679 TI - The natural history of hypertrophic cardiomyopathy. PMID- 3048681 TI - The hemodynamic evaluation in hypertrophic cardiomyopathy: systolic and diastolic dysfunction. PMID- 3048680 TI - Clinical presentation and noninvasive evaluation of the patient with hypertrophic cardiomyopathy. PMID- 3048682 TI - Medical therapy of end-stage congestive and ischemic cardiomyopathy. PMID- 3048684 TI - Patient selection and results of cardiac transplantation in patients with cardiomyopathy. AB - Cardiac transplantation for the treatment of end-stage congestive heart failure has been shown to be of benefit regardless of the etiology. With few exceptions, the evaluation of patients with end-stage heart failure is the same, regardless of the etiology. In those with cardiomyopathy not as a result of CAD, special attention must be given to exclude secondary causes of cardiomyopathy such as amyloidosis, hemochromatosis, and sarcoidosis, as well as generalized systemic illnesses that may also involve the heart, either secondary or hereditary, because special consideration must be given to these patients on a case-by-case basis to determine that there is no general systemic involvement of the illness that would preclude satisfactory rehabilitation after transplantation. Before cardiac transplantation becomes widely available, there must be a greater number of donor hearts, the lack of which now severely limits the number of transplants performed in comparison with the estimated need.66 Additionally, more effective and specific immunosuppressive agents must be identified in order to reduce the incidence of rejection, infection, and accelerated atherosclerosis that now limits the longevity of transplant recipients. Furthermore, the ideal immunosuppressive agent should be associated with fewer side effects than those currently available. The emotional and economic burdens placed on the patient, the family, and society must be balanced against the benefits generated by the procedure. Despite these limitations, cardiac transplantation continues to offer hope for the terminally ill patient, which must be tempered by an understanding of the real limitations of transplantation. PMID- 3048683 TI - Congestive heart failure and ventricular arrhythmias. PMID- 3048685 TI - Overview and classification of the cardiomyopathies. PMID- 3048686 TI - Diagnostic considerations in the adult patient with cardiomyopathy or congestive heart failure. PMID- 3048687 TI - Pathophysiology of congestive heart failure secondary to congestive and ischemic cardiomyopathy. AB - Dilated cardiomyopathy, owing to any cause, usually culminates in the clinical syndrome of congestive heart failure. Heart failure is characterized by exertional dyspnea and fatigue, but the precise mechanisms that produce these symptoms are still not clear. Sodium retention occurs early in heart failure, but this disturbance is dynamic in nature and is not always present in the patient. The mechanism of early salt and water retention in heart failure is not defined. Gross edema and ascites occur much later, undoubtedly owing to the convergence of a number of factors. The peripheral adaptations to heart failure include activation of the renin-angiotensin system and the sympathetic nervous system, and the release of AVP. The result is an increase in preload with a resultant increase in stroke volume for some patients, but the price is paid in the form of heightened impedance to ejection and circulatory congestion. The sympathetic nervous system disturbances in heart failure are striking, as disturbances in both circulating and myocardial NE levels are consistently found. Vasorelaxant and natriuretic hormones, as well as certain prostaglandins, may be released in an attempt to offset excessive "compensatory" pressor-sodium retentive mechanisms, but the net result seems to be excessive peripheral vasoconstriction and a downward spiral of deterioration in many patients. One would hope that an unraveling of the complex pathophysiology of heart failure would lead to therapy that would change the natural history of the disease. The results of the first V HeFT trial give room for cautious optimism in this regard. PMID- 3048688 TI - The clinical presentation and laboratory evaluation of congestive and ischemic cardiomyopathies. PMID- 3048689 TI - Pathology of cardiomyopathies. PMID- 3048690 TI - Effect of cardiopulmonary bypass on circulating concentrations of leucocyte elastase and free radical activity. AB - Circulating concentrations of leucocyte elastase and free radical activity were measured in 11 adults undergoing cardiopulmonary bypass. In all patients the bypass procedure was associated with pronounced changes in plasma elastase concentrations, and peak enzyme concentrations correlated closely with the duration of bypass (r = 0.91, p less than 0.001). Serial measurement of octadeca 9, 11-dienoic acid, a non-peroxide marker of free radical activity, showed significant changes only in the plasma free fatty acid fraction, suggesting a direct relation to the action of heparin rather than to the bypass procedure as such. These studies support the hypothesis that neutrophil activation plays a central role in the organ dysfunction that may complicate cardiopulmonary bypass and suggest that elastase release rather than free radical generation may be the appropriate marker of the event. PMID- 3048691 TI - Assessment of severity of cardiac rejection in heterotopic heart transplantation using indium-111 antimyosin and magnetic resonance imaging. AB - Seven canine donor hearts in which atrial septal defect and tricuspid regurgitation had previously been produced were heterotopically transplanted into the recipients' chest cavities. Indium-111 antimyosin myocardial imaging of the excised heart was performed using a scinticamera. Magnetic resonance imaging was also performed and the T2 relaxation time calculated. Subsequently, these data were correlated with pathological findings, which indicated the degree of rejection. Indium-111 antimyosin uptake was high in moderate and severe rejection, but the T2 relaxation time was prolonged even in mild rejection. Thus indium-111 antimyosin uptake was specific, and the T2 relaxation time was sensitive, for detecting the severity and extent of cardiac rejection. Although ex vivo experimental results have been reported, these new methods allow characterisation and accurate evaluation of myocardial tissue undergoing cardiac rejection. PMID- 3048693 TI - [Effect of fish oils on plasma lipids, glucose tolerance and insulin secretion in persons with endogenous hypertriglyceridemia]. PMID- 3048692 TI - Prostaglandin I2 analogue and propranolol prevent ischaemia induced mitochondrial dysfunction through the stabilisation of lysosomal membranes. AB - Leakage of lysosomal enzymes is associated with irreversible cellular damage. To determine the effect of prostaglandin I2 analogue and propranolol on the ischaemic myocardium in relation to changes in lysosomal integrity 26 anaesthetised mongrel dogs were divided into three treatment groups and subjected to 2 h coronary occlusion. In the control group (n = 12) physiological saline was infused throughout the experiment. In the prostaglandin I2 analogue group (n = 7) the prostaglandin I2 analogue, OP-41483-alpha-CD;5(E)-6-Deoxa-6,9 alpha-methylene 15-cyclopentyl-16,17,18,19,20-pentanor-PGI2. alpha-cyclodextrin clathrate (5 ng.kg-1.min-1) was infused from 25 min before occlusion until the end of the experiment. In the propranolol group (n = 7) propranolol (0.3 mg.kg-1) was injected for 10 min 25 min before occlusion. Two hours after occlusion mitochondria were prepared from both ischaemic and non-ischaemic areas in each group and their function measured polarographically with succinate as substrate. Fractionation of myocardial tissue from both non-ischaemic and ischaemic areas was performed and the activities of lysosomal enzymes (N-acetyl-beta glucosaminidase; beta-glucuronidase) were measured. In the control group, mitochondrial function in the ischaemic area was reduced compared with that from the non-ischaemic area. The activities of both lysosomal enzymes were increased significantly in the supernatant fraction obtained from the ischaemic area compared with those for the supernatant from the non-ischaemic area. The administration of prostaglandin I2 analogue or propranolol not only prevented the leakage of lysosomal enzymes but also maintained mitochondrial function.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3048694 TI - [Steady-state plasma levels of piroxicam of 4 different origins and relation to clinical effects in the treatment of rheumatoid arthritis]. PMID- 3048696 TI - [Problems surrounding substitution therapy with insulin]. PMID- 3048695 TI - [The history of urology at the Prague Medical School]. PMID- 3048697 TI - [Endothelial vasodilating factor]. PMID- 3048699 TI - [The Czech Medical Association during the Nazi occupation]. PMID- 3048698 TI - [Statistics from the last epidemic of bubonic plague in Prague 1713-1714]. PMID- 3048700 TI - Classification and diagnostic criteria for headache disorders, cranial neuralgias and facial pain. Headache Classification Committee of the International Headache Society. PMID- 3048701 TI - Extremely conserved histone H4 N terminus is dispensable for growth but essential for repressing the silent mating loci in yeast. AB - Yeast histone H4 function was probed in vivo by deleting segments of this extremely conserved 102 amino acid protein. Deletions in the hydrophobic core of H4 are lethal and block chromosomal segregation. In contrast, deletions at the hydrophilic N terminus (residues 4-28) and C terminus (residues 100-102) are viable. However, N-terminal deletion alters normal chromatin structure and lengthens the cell cycle, especially G2. Surprisingly, removal of the H4 N terminus also derepresses the silent mating type loci, HML alpha and HMRa, disrupting mating. This activation is specific since other regulated genes (GAL10, PHO5, CUP1) are repressed and induced normally in these cells. Deletions of the hydrophilic N termini of H2A or H2B do not show this effect on mating. These experiments allow us to define a unique H4 function that is not shared by other histones (H2A and H2B). PMID- 3048702 TI - Two conserved domains of yeast U2 snRNA are separated by 945 nonessential nucleotides. AB - Yeast U2 snRNA (1175 nucleotides) is six times larger than its mammalian counterpart (188 nucleotides). Using deletion analysis, we show that the molecule can be divided into three phenotypically distinct domains. As expected, the highly conserved 5' domain (approximately 120 nucleotides) is absolutely essential for viability. Surprisingly, however, deletion of the central 945 nucleotides has no effect on growth rate. In contrast, removal of sequences in the 3' terminal 110 nucleotides results in low numbers of slow-growing colonies; these cells contain U2 with altered 3' ends. This domain can be folded into a secondary structure that strongly resembles the 3' terminal stem-loop IV of human U2. We conclude that yeast U2 contains two functionally important elements. While the 5' domain is known to be directly involved in the splicing reaction, the 3' domain may function primarily in the generation of stable small nuclear ribonucleoprotein particles. PMID- 3048703 TI - Sequence requirements for nucleolar localization of human T cell leukemia virus type I pX protein, which regulates viral RNA processing. AB - The posttranscriptional regulator (p27x-III) of human T cell leukemia virus type I (HTLV-I) is located predominantly in the cell nucleolus. A highly basic amino terminal sequence (NH2-Met-Pro-Lys-Thr-Arg-Arg-Arg-Pro-Arg-Arg-Ser-Gln-Arg-Lys Arg-Pro-Pro -Thr- Pro) in this protein, when fused to the amino termini of beta galactosidase and p40x of HTLV-I, acts as an autonomous signal capable of directing the hybrid proteins to the cell nucleolus. PMID- 3048705 TI - Role of laminin A chain in the development of epithelial cell polarity. AB - Kidney organ culture was used to study the conversion of embryonic mesenchymal cells into a polarized, differentiated kidney epithelium. We examined the expression of laminin, a basement membrane glycoprotein, during this conversion. The B chains of laminin were constitutively expressed, whereas the appearance of the A chain of laminin was dependent on embryonic induction and coincided with the onset of cell polarization. Antisera against the carboxy-terminal end of laminin inhibited polarization but did not affect the developmental events that precede polarization. Antisera against N-terminal parts of laminin failed to inhibit morphogenesis. Since the fragments at the carboxy-terminal end contain parts of the A chain, we suggest that the appearance of this chain is fundamental for initiation of cell polarity. PMID- 3048704 TI - Cloning and functional expression in bacteria of a novel glucose transporter present in liver, intestine, kidney, and beta-pancreatic islet cells. AB - The well-characterized erythrocyte glucose transporter is also expressed in brain, adipocytes, kidney, muscle, and certain transformed cells, but not in liver, intestine, or the islets of Langerhans. Using as probe a cDNA encoding the rat brain glucose transporter, we isolated from a rat liver cDNA library a clone encoding a protein 55% identical in sequence to the rat brain transporter, and with a superimpossible hydropathy plot. We expressed this protein in an E. coli mutant defective in glucose uptake; the protein was incorporated into the bacterial membrane and functioned as a glucose transporter. This new transporter is expressed in liver, intestine, kidney, and the islets of Langerhans; immunofluorescence analysis showed that it is present in the plasma membrane of the insulin-producing beta cells. Insulinoma cells express, inappropriately, the erythrocyte glucose transporter, and we suggest that this may be related to their inability to secrete insulin in response to elevations in glucose. PMID- 3048707 TI - Mechanisms of color vision. AB - We review the physiological and psychophysical research on mechanisms of color vision. Psychophysical work has led to the formulation of explicit theories of the early stages of color vision. The principal postulates of these theories have been confirmed by physiologists (e.g., the existence of three classes of receptors and second-stage mechanisms in which the signals from these receptors are compared), but some important features of the psychophysical scheme have found limited physiological support. One such issue is the absence of the unitary "achromatic" mechanism required by psychophysicists. We know a good deal less about the chromatic analyses that occur beyond these early stages. Although physiologists have devoted much effort to the study of cortical mechanisms, little of this work has been guided by clear ideas of the tasks performed by them. The provision of color constancy and the ability to segment scenes are perhaps the foremost concerns of chromatic mechanisms, and recent psychophysical work bearing on these problems offers physiologists clearer guidance on what to seek with their electrodes. PMID- 3048706 TI - In vivo analysis of the mechanisms responsible for strong transcriptional polarity in a "sense" mutant within an intercistronic region. AB - We have studied a very unusual strong polar mutant in the intercistronic barrier between the second (hisD) and third (hisC) cistrons of the histidine operon of Salmonella typhimurium to obtain further insights into the molecular mechanisms leading to transcription termination within cistrons. We have performed a detailed transcriptional analysis in vivo and have found that the his mRNA in this polar mutant is reduced in size as a result of premature termination of transcription at a cryptic Rho-dependent site within the proximal region of the hisC cistron. PMID- 3048708 TI - [Immunofluorescence determination of antibodies in men with chlamydial urethritis]. PMID- 3048709 TI - [Transvaginal aspiration of follicular fluid using a sonographic vaginal probe in an in vitro fertilization and embryo transfer program]. PMID- 3048710 TI - [Vaginal sonography--new horizons in diagnosis]. PMID- 3048712 TI - [Personal experience with uterine sutures in cesarean section over a 12-year period]. PMID- 3048711 TI - [Therapeutic transplantation of a skin graft from the partner into an infertile female patient]. PMID- 3048713 TI - [Ultrasound in the detection of residua after labor, abortion and induced abortion]. PMID- 3048714 TI - [Transabdominal amniocentesis during continuous ultrasonic monitoring]. PMID- 3048716 TI - [Imaging of pathologic masses based on type A ultrasonography in ophthalmology]. PMID- 3048715 TI - [Possible use of interventional echography in pregnancy]. PMID- 3048717 TI - [The Ophthascan B ultrasonoscope in clinical practice]. PMID- 3048718 TI - [Tapioca melanoma of the iris]. PMID- 3048719 TI - [Cyclosporin A in corneal transplantation]. PMID- 3048720 TI - [Arlt's years in Prague]. PMID- 3048721 TI - [80 years' activity of the Ophthalmology Department of the hospital in Prostejov]. PMID- 3048722 TI - [Atherosclerosis and childhood]. PMID- 3048723 TI - [Czech and Slovak pediatricians as authors]. PMID- 3048724 TI - [Diabetes mellitus in children and adolescents]. PMID- 3048725 TI - [Pulmonary changes in collagenoses]. PMID- 3048726 TI - [IgA nephropathy]. PMID- 3048727 TI - [The beginnings of comprehensive pediatric care in Ostrava]. PMID- 3048728 TI - Treatment of infections in non-neutropenic patients with cancer, AIDS, or renal transplant using ciprofloxacin. AB - Infective complications not only remain the major factor in preventing a large proportion of cancer patients from achieving a complete remission, but also greatly influence the outcome of immunosuppressed transplant recipients and are a prominent problem in subjects affected by AIDS. We report the results of 73 patients, 54 males and 19 females with a mean age of 61.4 +/- 15.1 years affected by solid cancer (64 pts), AIDS (6 pts) or with kidney transplant (3 pts), treated with ciprofloxacin 250 mg bid (21 pts) or 500 mg bid (52 pts) for respiratory tract infections (41 cases), urinary tract infections (22), septicemia (5) or other infections (5). The mean course of therapy was 9.6 +/- 5.7 days and led to a complete resolution of symptoms in 66 (90.4%) patients, improvement in 6 (8.2%), while the clinical picture was unaffected in 1 (1.4%). Candida superinfection occurred in one case and only four patients experienced side effects. Ciprofloxacin results to be an effective antibacterial agent in a high risk population. PMID- 3048730 TI - [Spontaneous pneumothorax: surgical aspects]. AB - The authors illustrate the cases of spontaneous pneumothorax observed in their institute along with the diagnostic and therapeutic aspects. In accord with the experience of others as reported in the literature, they stress the importance of a precise, immediate diagnosis for the purposes of adopting the treatment of choice: in the presence of closed, partial forms a conservative approach is to be preferred, whereas in open and/or hypertensive cases an aspirative thoracic drainage must be provided without delay. Aggressive surgical treatment, with resection of the bullous lesions responsible for bronchopleural fistulas, will resolve the condition and should be implemented after a reasonable waiting period with aspirative drainage, or even, electively, in the first instance, in the presence of multiple relapses. In any event, the authors stress the need to avoid uncertainty and delay in view of the youthful age of most of the subjects affected and the potential danger of the complications of this apparently banal disease. PMID- 3048729 TI - [Biomechanico-clinical study of gunshot wounds (general problems--II)]. AB - Whenever the surgeon finds himself face to face with a wound (probably this is the only opportunity for a meeting between physician and pathology which seems to be able to leave the "illness" on one side, almost forgotten, as it were), even when immersed in routine, he can hardly help making a number of considerations of a general nature, to which the sentence above in brackets is not entirely extraneous. In practice, we cannot help asking ourselves an apparently simple, almost banal, question: what exactly is trauma? This triggers off a whole series of secondary queries, such as, for instance, what the relationship is between trauma and classical pathology? In the first place, it should be pointed out that "traumatic" pathology is undoubtedly the only instance of pathology in which, as a rule, at least at the outset, one can justifiably talk about the "isolated" role of what can certainly be regarded as an out-of-body factor. If, then, we consider the specifically morphological and pathophysiological aspects of the period subsequent to the traumatic insult, we find ourselves in an even more embarrassing position: we are faced with irreparably devastated organ and body structures, or with a situation which is already on the way to convalescence. One last alternative is that the traumatic insult is merely a memory, a key finding in the case history, a past reality which to all intent and purposes has ceased to exist, and we are faced with extremely complex clinical pictures which we tend to label as complications. A few examples by way of explanation: shock, adult respiratory distress syndrome (ARDS), stress ulcer, acute post-traumatic cholecystitis, haemorrhagic pancreatitis, and problems caused by resolving the hypovolaemia-ischaemia situation and by implementing reperfusion (oxygen radicals). Trauma favours - and surgeons concerned with organ transplants are well aware of this - the only possibility of death which, perhaps with a grain of excessive optimism, we may even accept as fruitful, in that it occurs without all the destructive deterioration involved in the process of dying. The above consideration probably plays a major role in our attitudes of almost fatalistic resignation towards the youthful victims of trauma.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3048731 TI - [Primary malignant lymphomas of the gastrointestinal system]. AB - The Authors describe their experience regarding eight cases of primary malignant gastrointestinal lymphomas. After having briefly reexamined the epidemiology and the problems coming from the discordances that still exist on various systems of classification and of staging, they put in evidence that it is not always possible to reach a sure preoperative diagnosis on such pathology, and they explain some considerations about the diagnosis, the therapy and the prognosis. PMID- 3048732 TI - [Usefulness of Doppler echography in the diagnosis of upper thoracic outlet syndrome]. AB - The Authors emphasize the importance of the Doppler-sonography for the diagnosis of the outlet thoracic syndrome with our personal casuistry. PMID- 3048733 TI - [Diagnostic usefulness of Doppler sonography in the study of cerebrovascular pathology]. AB - The Authors point out the doppler usefulness for the study of obstructive cerebrovascular pathology specially of the carotids with personal short casuistry. PMID- 3048734 TI - [Mediastinal thymoma (presentation of 2 cases)]. AB - The Authors report on two cases of mediastinal thymoma. Clinical and anatomicopathological aspects are discussed. PMID- 3048735 TI - [Injuries of the bladder and urethra in children. Commentary apropos of a multicenter study of 187 cases]. AB - 187 cases of iatrogenic urethral and vesical traumatism from 25 pediatric surgery departments are analysed. Results and especially percentages of complications are compared with literature. The most important of them are stenosis. The authors propose a management for study, treatment and overlooking of these children who are often polytraumatized. PMID- 3048736 TI - [Fractures of the pelvis and associated bladder-urethral rupture]. AB - We report 82 cases of pelvic fractures associated with vesico-urethral ruptures. Pelvic fractures were 23 simple fractures, 5 bilateral obturator foramen's margin fractures, 30 pubic dislocations sometimes associated with other fractures and 24 complex fractures. Urological injuries were 64 urethral ruptures, 13 bladder ruptures and 5 vesicourethral injuries. These well documented cases allow us to study relations between bone and visceral injuries as well as result's quality in relation to treatment. From this study a treatment schedule has been deduced insisting upon the fact these bone and visceral injuries must be treated immediately and if possible in a one time procedure. The repairs time schedule depends upon the fracture's type from which benefit can be taken out for the urethral access. PMID- 3048737 TI - [Injuries of the rectum in children. 79 cases]. AB - The observations of 79 patients with rectal or ano-rectal lesions were studied in 15 centers, on a 25 years period. The etiology of the lesions was: 23 patients which suffered from traffic road accident, 39 patients which suffered from impaling, 10 patients with lesions caused by foreign bodies, 6 patients which suffered sexual violence and 1 patient with other causes. The primary results were: 2 patients who died, and 27 patients who suffered from other complications. 19 of them were of infectious nature and the other 6 suffered from urethral rectal or vaginal-rectal fistula. The secondary results showed that 28 patients developed early complications and 1 patient developed late complications. The majority of the complication were: 9 anal stenosis and 11 were anal sphincter incompetence. The analysis of the results showed the factors implicated in the appearance of these complications, which were: the initial unknown nature of the lesions in the polytraumatised patients, the nature of the lesions, the associated lesions of the sphincter or the perineum, the late diagnosis and the factors which increase the risk of an early infection. PMID- 3048738 TI - [Injuries of the testis in children]. AB - Thirty three blunt testicular injuries have been observed within the frame work of the French Society of Pediatric Surgery. Four histological stages are described. Stage 1: testis contusion: 17 cases. Stage 2: rupture and retraction of the albuginea: 10 cases. Stage 3: complete testis rupture: 4 cases. Stage 4: testis fragmentation: 2 cases. Diagnosis remains clinical in spite of the recent progress of ultrasonography. After surgical treatment results show: 9% castration, 25% testis atrophia, that is to say one testis out of three is lost after traumatism. The hormonal function was not affected. 2 Testis biopsies allowed us to study spermatogenesis. There is no proof of astheno or azo or teratospermia after childhood traumatism. Sterility by autoimmunization has been mentioned only in adults. PMID- 3048739 TI - [Lacerations of the perineum in children]. AB - 24 cases of disruption of the perineum from 8 Centres of Pediatric Surgery in France, were studied. The patients were aged between 4 months and 13 years, with an average age of 6.7 years, and 70% were boys. The cause of the injury was crushing in 13 cases. The associated were complex. Fractures of the pelvis (18) and associated multiple fractures (13), as were as profuse pelvi-peritoneal haemorrhages arising from fractures (3) or lower limb amputations (3) determined the care of associated soft-tissue injuries. Disruptions of the skin were almost constant and displayed a high rate of sepsis. There were noted lesions of the ureter (9), the vagina (5), the anus and rectum (17), the ano rectal sphincter (10) and the penis (1). The care of each of these lesions followed the usual practices of Reparative Surgery specific to each organ, but was also adapted to each case. In effect, disruptions of the perineum take place in a context of serious polytrauma, where the hierarchy of urgency determines the choice of therapeutic attitudes. PMID- 3048740 TI - [Value of magnetic resonance imaging in the follow-up of children operated for portal hypertension]. AB - The authors report their experience concerning nuclear magnetic resonance imaging (NMR) in 7 children. They emphasize its importance in the post-operative study of portal hypertension. All those children got surgically created porto-systemic shunts. NMR imaging is easy, safe and trustable. It is able to visualise the shunt itself and to demonstrate the patency of this shunt. So, it is interesting when the usual controls have failed, particularly abdominal ultrasonography. PMID- 3048741 TI - [Ureteropelvic junction syndromes: antenatal diagnosis]. AB - Seventy-four hydronephrosis by uretero-pelvic obstruction, discovered by antenatal real-time ultrasound, have been managed on sixty-one children. Three groups, with a specific management, are distinguished. The minimal forms have been only watched over (15 cases). The average forms have been operated (49 cases); considering the results (3 immediate complications and 2 early stenoses), it seems better to wait for 4 months to operate and to place a drain in renal cavities. In major forms (silent kidney on IVP), the diagnosis and therapeutic management use the percutaneous nephrostomy (10 cases). The interest of antenatal diagnosis of uretero-pelvic obstructions is to allow to operate, not only before the renal deterioration emphasizes, but principally before septic complications. PMID- 3048743 TI - [Calcified thrombosis of the inferior vena cava in children]. AB - A new case of calcified thrombus in the Inferior Vena Cava is described, and added to the 19 previously reported cases in literature. The etiology of these calcified thrombi is unknown, but a precise diagnostic an therapeutic approach is now well established. On plain X-ray of the abdomen, the right paravertebral bullet-shaped calcification is diagnostic. No further investigations are necessary, except in the neonatal period, when the thrombus is incompletely calcified, suggesting neuroblastoma or adrenal hemorrhage. Definitive diagnosis, in that case, can be made by abdominal ultrasonography. In the current state of our knowledge, the clinical tolerance of this anomaly is excellent. No specific treatment appears to be necessary. PMID- 3048742 TI - [Artificial urinary sphincter in the treatment of neurogenic bladder in children]. AB - Over a two years period, the model AS 800 artificial urinary sphincter was implanted in 10 children (mean 14 years old) with incontinence due to a neurogenic bladder (7 spina bifida; 3 sacral agenesis). Goal of continence was achieved in 100% of 7 boys and 3 girls with a follow up between 6-32 months (mean 17 months). Patients selection was rigid and incontinence have failed to response at pharmaceutic manipulation (6) intermittent catheterisation (5) and previous classical surgery enterocystoplasty (2), Young-Dees procedure (1). Age minimum was 8 years. All sphincters were implanted around the bladder neck. Associated surgical techniques were performed in 4 cases: 2 detubularized enterocystoplasties, 1 Teflon injection for vesico ureteral reflux, 1 closure of previous cystostomy. Sphincterotomy was never done. 5 patients are submitted post implantation to intermittent catheterisation without any problem. 2 patients required one revision. Nocturnal desactivation is used in 2 cases. Authors focus on the necessity of correcting previously or at the time of implantation bladder compliance and vesico ureteral reflux. PMID- 3048744 TI - [Unilateral hematocolpos in utero-vaginal duplication associated with homolateral renal agenesis. Apropos of 2 clinical case reports]. AB - This is a report of two Cases of unilateral hematocolpos with ipsilateral renal agenesis in a bicornuate uterus with unilateral imperforate vaginal. The Symptoms, operative findings and surgical procedure are described. It is exceptional in the neonatal period to find an Hydro-Hematocolpos associated with complex genitourinary malformations. Easy diagnosis of Hydro-Hematocolpos and its effect on upper urinary tract are made by ultrasonography and intravenous urography. PMID- 3048745 TI - The production of myeloid blood cells and their regulation during health and disease. AB - The regulation of myelopoiesis in vivo most likely entails a complex set of interactions between cell-derived biomolecules and their target cells: hematopoietic stem and progenitor cells and accessory cells. Stimulating and suppressing factors have been characterized through in vitro studies, and their mechanisms of action in vitro and in vivo have begun to be elucidated. Among those factors being studied are the hematopoietic colony-stimulating factors (CSF): interleukin-3 (multi-CSF), granulocyte-macrophage-CSF, granulocyte-CSF, and macrophage-CSF; other molecules include erythropoietin, B-cell-stimulating factor-1, interleukin-1, interleukin-2, prostaglandin E, leukotrienes, acidic ferritins, lactoferrin, transferrin, the interferons-gamma, -alpha, and -beta, and the tumor necrosis factors-alpha and -beta (lymphotoxin). These factors interact to modulate blood cell production in vitro and in vivo. The proposed review characterizes these biomolecules biochemically and functionally, including receptor-ligand interactions and the secondary messengers within the cell which mediate their functional activity. The production and action of the molecules are described under conditions of hematopoietic disorders, as well as under normal conditions. Studies in vitro are correlated with studies in vivo using animal models to give an overall view of what is known about these molecules and their relevance physiologically and pathologically. PMID- 3048746 TI - Radioimmunodetection of cancer with monoclonal antibodies: current status, problems, and future directions. AB - Early studies of immunoscintography with affinity-purified 131I-labeled polyclonal antibodies reactive against oncofetal antigens such as carcinoembryonic antigen (CEA) were moderately successful in detecting metastatic colorectal carcinoma. However, because of low tumor to background ratios of isotope, background subtraction techniques using 99Tc-labeled albumin were required to visualize small lesions. Antisera were often of low titer and lacked specificity. These problems could be overcome for the most part following the development of highly specific monoclonal antibodies (MoAb) against a variety of tumor-associated antigens. A number of clinical trials using 131I- or 111In labeled MoAb to image tumors have demonstrated successful localization without the use of subtraction techniques. Variables limiting the usefulness of murine MoAb for diagnosis have included increased localization in liver and spleen, tumor vascularity and heterogeneity of antigen expression, and development of human antimurine globulins. Methods to overcome some of these problems are discussed. Radiolabeled MoAb appear useful as an adjunct to conventional diagnostic techniques both as a means to predict which antibodies might be useful for treatment and, in select patients, as a basis for treatment decisions. PMID- 3048747 TI - [A complete form of basocellular nevomatosis manifested by exophthalmia]. PMID- 3048748 TI - [Peters' syndrome: etiopathogenesis and treatment. Apropos of a case]. PMID- 3048749 TI - [Tolerance for artificial silicone lenses]. PMID- 3048750 TI - [Technic of epikeratoplasty]. PMID- 3048751 TI - [Ophthalmic migraine and arterial cervico-cephalic malformations: apropos of a case report]. PMID- 3048752 TI - Anticholinergic drugs and anaesthesia. PMID- 3048753 TI - The effect of pH adjustment of bupivacaine on onset and duration of epidural anaesthesia for caesarean section. AB - Previous studies have reported that elevation of the pH of local anaesthetics results in more rapid onset of action, with enhanced quality and duration of block. This study investigated the effect of pH adjustment of 0.5 per cent bupivacaine immediately prior to epidural anaesthesia for Caesarean section. Addition of 0.1 ml of 8.4 per cent sodium bicarbonate to 20 ml of 0.5 per cent bupivacaine consistently raised the pH of the local anaesthetic from 5.49 to 7.04 (mean values). One hundred patients, presenting for elective Caesarean section under epidural anaesthesia participated in the study. Forty patients received epidural anaesthesia, using pH-adjusted 0.5 per cent bupivacaine, in a dosage adequate to produce block to the T4 level. A control group of 40 patients received the standard commercial preparation of 0.5 per cent bupivacaine. A further ten patients in each group received epidural anaesthesia using 0.5 per cent bupivacaine with the addition of 1:400,000 epinephrine, to study the effect of epinephrine on pH adjustment of the local anaesthetic. Elevation of the pH of the local anaesthetic significantly increased the speed of onset of action from 6.4 minutes to 3.2 minutes and the time to peak effect from 24.8 minutes to 18.1 minutes, while the duration of anaesthesia was increased from 124.8 minutes to 147.3 minutes. The time to S2 segment blockade was also shortened from 13.5 to 8.6 minutes. Addition of 1:400,000 epinephrine to the local anaesthetic did not influence the effect of pH adjustment. Maternal and umbilical cord plasma levels of bupivacaine were not affected by pH adjustment of the local anaesthetic, while MV/UV and UA/UV ratios were unaltered. PMID- 3048755 TI - The literature of anaesthesia: what are we learning? AB - In an effort to identify the types of articles published in anaesthesia literature, a stratified random sample of articles published in North America between 1977 and 1986 was analyzed (N = 571). Human studies constituted 63 per cent of the total, with case reports and case series constituting over half. Study designs classes as descriptive in nature were remarkably rare in the anaesthesia literature, with prevalence and case-control studies constituting 0.8 and 3.3 per cent of the total respectively. Cohort studies (7.8 per cent), non randomized intervention studies (12.8 per cent), and randomized controlled trials (17.8 per cent) were more numerous, but many suffered major contamination of experimental design. Frequently identified concerns in assessing the applicability of a given study to general anaesthetic practice were a bias induced by selection of the study subjects, application of the results from tertiary care hospitals to community hospitals, and contamination of the study protocol. These factors were identified as present in the majority of articles. The results suggest that growth of the specialty of anaesthesia is constrained by the narrow spectrum of study designs, as well as major problems affecting generalizability of the published results. PMID- 3048754 TI - Anaesthesia for cardioversion: a comparison of propofol and thiopentone. AB - Propofol (2,6-diisopropylphenol) 2.5 mg.kg-1 IV was compared with thiopentone 5 mg.kg-1 IV as an induction agent in anaesthesia for elective cardioversion. Thirty-five patients (ASA physical status II-III) with atrial fibrillation were included. Thirty patients were randomized to receive propofol or thiopentone. Five patients were treated twice during the study period and anaesthetized with both agents (the first treatment according to the random order and the second with the agent not used on the first occasion). The induction characteristics and the haemodynamic response for propofol and thiopentone were similar. The success rate of cardioversion did not differ between the groups. Recovery times were shorter after propofol than after thiopentone with respect to ocular muscle balance, central integration and subjective sedation of patients. The incidence of side-effects did not differ between the groups. None of the patients reported any awareness during the procedure. All five patients treated twice (with both agents) assessed the anaesthetic procedure with propofol as being more pleasant than that with thiopentone. PMID- 3048756 TI - Negative pressure pulmonary oedema secondary to airway obstruction in an intubated infant. AB - We report the case of a healthy one-month-old male infant who underwent an uneventful endotracheal anaesthetic for hernia repair. During transport to the recovery room (a less than 30 second trip), the endotracheal tube in the spontaneously breathing infant became obstructed, possibly due to impaction of the tip in the right main bronchus. Restoration of the airway was followed by fulminant pulmonary oedema. Several days of vigorous respiratory and pharmacologic therapy were required for resolution of the infant's respiratory distress. We review other reported cases of acute airway obstruction associated with pulmonary oedema in children and briefly describe the proposed mechanisms. The difficulties of gauging proper endotracheal tube depth in the infant are noted. This case report demonstrates the importance of continuous monitoring during patient transport to the recovery room. PMID- 3048757 TI - Anaesthesia for phaeochromocytoma. PMID- 3048758 TI - In vivo and in vitro responses to quinine and quinidine of Plasmodium falciparum. AB - A total of 66 Thai children with uncomplicated falciparum malaria were treated orally with regimens of either quinine or quinidine. Radical cures were observed in 85% (28 of 33) of the children who received quinine and in 88% (29 out of 33) of those who received quinidine. Treatment failures in both groups were RI responses.The mean trough level of quinidine (10 mumol/l) was about 2.5-times less than that of quinine (25 mumol/l). The electrocardiograms of the two treatment groups differed significantly in that there was an acute prolongation of the QT(c) interval in 56% of those who received quinidine compared with 21.0% of those given quinine. In vitro assays of the pretreatment drug susceptibilities of the isolates of Plasmodium falciparum indicated that the mean minimum inhibitory concentration (MIC) for quinidine (1.44 mumol/l) was about half that for quinine (3.02 mumol/l). Although both drugs are equally effective, quinine is recommended for treatment of multidrug-resistant malaria in paediatric patients, primarily because of the cardiac effects produced by quinidine. PMID- 3048759 TI - Effectiveness of amodiaquine, sulfadoxine-pyrimethamine, and combinations of these drugs for treating chloroquine-resistant falciparum malaria in Hainan Island, China. AB - The study was carried out in 1985-86 in Hainan Island where Plasmodium falciparum is resistant to chloroquine. Fifty cases of falciparum malaria were treated with 1800 mg amodiaquine for 3 days: the cure rate was 65.3%, and the mean time to clear fever and asexual parasitaemia was 30.7 and 60.3 hours, respectively; 34.7% of cases showed RI or RII recrudescence, and one patient's temperature did not come down to normal within 7 days.Twenty-one cases were treated with sulfadoxine pyrimethamine (1500 mg and 75 mg, respectively): 19 were cured, I showed RI and another had an S or RI response; the mean time for fever control was 56.1 hours.Fifty cases were treated with amodiaquine plus sulfadoxine and 49 received amodiaquine plus sulfadoxine-pyrimethamine: the cure rate was 97.9% and 100%, respectively; the mean time for fever clearance was 25.0 and 25.7 hours and for parasite clearance 57.1 and 52.8 hours, respectively. These drug combinations gave much better results for cure and for symptom control than amodiaquine or sulfadoxine-pyrimethamine alone, and may be considered for treatment of chloroquine-resistant falciparum malaria. PMID- 3048761 TI - A quantitative approach to recommendations on malaria prophylaxis. AB - In order to develop recommendations for malaria prophylaxis, a quantitative method is needed to balance the risk of Plasmodium falciparum malaria infections against the toxicity of antimalarial drugs. Using decision analysis, we estimated the expected mortality associated with three alternative regimens of prophylactic drugs for visitors to three areas with different risks of infection with chloroquine-resistant P. falciparum. The model used took into account the risks of malaria and of adverse reactions to antimalarial drugs. Estimates of the parameters used in the analysis were based on observations made on U.S. travellers. Reducing the risk of malaria infection was found to have a far greater impact on lowering the expected mortality than that of increasing the chemoprophylactic efficacy of the drugs used, thereby emphasizing the need for travellers to use anti-mosquito measures in malarious areas. The analytical method described can be used to define optimal malaria prevention strategies. PMID- 3048760 TI - Circumsporozoite antibodies and falciparum malaria incidence in children living in a malaria endemic area. AB - In a case-control study we examined the association of Plasmodium falciparum circumsporozoite antibodies (anti-R32tet32) with subsequent P. falciparum infections. A study population of 140 children living in an endemic area was followed longitudinally for 25 weeks with weekly blood smears for malaria parasites and, once every two weeks, serum samples for circumsporozoite antibody determinations. From the malaria cases, antibody measurements occurring between two and six weeks prior to the onset of parasitaemia were utilized. For each case, two controls were selected. The results from 17 cases and 34 controls failed to show a statistically significant difference in antibody levels prior to the infection (P=0.07, one-tailed Student's t-test). However, 8 of the 17 cases had antibody present, indicating a level that was not protective against patent infection. PMID- 3048762 TI - Activity of a new antiemetic agent: alizapride. A randomized double-blind crossover controlled trial. AB - Alizapride is a methoxy-2-benzamide derivative three times more potent than its parent compound, metoclopramide, as an antagonist of apomorphine-induced emesis in dogs. The antiemetic activity of alizapride plus dexamethasone (DXM) was compared with that of placebo plus DXM in a randomized, double-blind, crossover study in cancer patients receiving cisplatin (DDP). Alizapride, given at the maximally tolerated dose of 4 mg/kg x 5, or placebo was given in a sequence determined by randomization during two successive, identical courses of antitumor chemotherapy. The antiemetic treatment was given 30 min before and 1.5, 3.5, 5.5, and 7.5 h after starting. DXM, in a dose of 12 mg, was given IV with the first administration of alizapride or placebo. A total of 39 patients completed the two courses of chemotherapy. The severity of gastrointestinal symptoms was influenced by previous treatment but not by the treatment sequence. Although our overall results suggest that alizapride does not add to the activity of DXM against DDP induced amesis, a statistically significant difference favoring alizapride plus DXM was found among patients with the lowest gastrointestinal tolerance to DDP: women, patients under 50 years of age, and patients pretreated with chemotherapy including DDP and non-DDP agents. Side effects consisted of orthostatic hypotension, which was symptomatic in two patients, and a single occurrence of severe extrapyramidal syndrome. We conclude that alizapride is more active than placebo when combined with DXM for DDP-induced emesis in patients at high risk of severe nausea and vomiting. The severity of the side effects in this study indicates that a dose reduction of alizapride might be appropriate for further studies. PMID- 3048764 TI - Expression of the oncogenes mil and ras abolishes the in vivo differentiation of mammary epithelial cells. AB - Three carcinoma-associated oncogenes, two of which have been strongly implicated in human mammary tumorigenesis, have been introduced into a novel mouse mammary epithelial cell line, EF43, that retains many differentiated functions. The effect of oncogene expression upon classical transformation parameters as well as parameters specific for mammary epithelial cells such as growth in three dimensional collagen matrices and the ability to repopulate the cleared mammary fat pad and to form alveolar structures in vivo has been investigated. Expression of v-myc in EF43 cells results in no obvious phenotypic changes, and does not confer tumorigenic potential upon the cells. Expression of v-Ha-ras confers upon EF43 cells the ability to grow rapidly, grow in an anchorage-independent manner, results in tumor formation in nude and syngeneic animals, abolishes their ability to repopulate the mammary gland and, instead, results in rapid induction of anaplastic tumors. The v-mil oncogene, an avian homolog of the mouse v-mht and human c-raf oncogenes, previously thought to be non-transforming in the absence of a co-operating oncogene, transforms EF43 cells, allowing them to grow in an anchorage-independent manner, form tumors in nude mice and abolishes their ability to repopulate the cleared mammary fat pad. In contrast to v-ras, however, the tumors arising from v-mil expression have a differentiated morphology, typical of adenocarcinomas. Thus, different oncogenes show varying degrees of inhibition of the differentiation of mammary epithelial cells in vivo. PMID- 3048763 TI - Resistance to cisplatin and analogues: mechanisms and potential clinical implications. AB - In view of the important role of cisplatin (CDDP) in cancer chemotherapy, the frequent occurrence of resistance to the drug is a major clinical problem. The main cause for unresponsiveness of a tumor to CDDP is thought to be cellular drug resistance, which may be caused by (1) a decreased uptake of CDDP, (2) an increase in metallothioneins, (3) an increase in glutathione and/or glutathione-S transferase, (4) increased DNA repair, or (5) increased tolerance to unrepaired lesions in DNA. Several mechanisms may be concomitantly operative. However, almost all data on CDDP resistance are derived from cell lines or experimental animal systems, and it is uncertain whether they are relevant for human tumors. Possible methods for overcoming CDDP resistance in cancer patients include the use of high-dose CDDP or carboplatin or of different formulations of platinum derivatives, the regional administration of CDDP, the inducement of hyperthermia, the depletion of glutathione by buthionine-S-R-sulfoximine (BSO), or the use of platinum analogues. The development of methods to detect and classify CDDP resistance in human tumor samples is urgently required for the development of modalities to overcome resistance. PMID- 3048765 TI - DNA binding by [2,5-14C]N-nitrosopyrrolidine in excision-repair proficient and deficient strains of Salmonella. Evidence for a major premutagenic adduct. AB - Little is known about the nature and possible genotoxic effects of the DNA adducts formed by N-nitrosopyrrolidine (NPYR) in whole animals. DNA binding in DNA isolated from [2,5-14C]NPYR-treated Salmonella was studied and attempts were made to monitor DNA adducts and correlate DNA binding with mutagenesis. NPYR was metabolized by hamster liver S-9 fraction in the presence of S.typhimurium TA1535 (uvrB-) or TA1975(uvrB+). DNA isolated from TA1535 contained about three times as much radioactivity as that isolated from TA1975, and NPYR-induced mutagenesis was several-fold higher in TA1535. The fraction of radioactivity incorporated into TA1535 was approximately 10(-5). Thermal hydrolysis of the 14C-containing DNA at neutral pH, followed by precipitation, released approximately 2/3 of the radioactivity into the supernatant. HPLC analysis of the supernatant revealed one major peak. This peak was absent in DNA from TA1975. Acid hydrolysis of the DNA precipitate after neutral hydrolysis released most of the residual radioactivity. Several small peaks were observed after HPLC analysis of the TA1535 acid hydrolysate or the TA1975 acid hydrolysate. These results demonstrate that NPYR is capable of binding to Salmonella DNA yielding one major product after hydrolysis and this DNA binding product appears to be repaired by the excision repair system. The fact that the major peak of radioactivity released from Salmonella is only found in the strain which is efficiently reverted by NPYR suggests that mutagenesis is dependent on the DNA modification leading to this peak. PMID- 3048766 TI - Influence of terbutaline on endotoxin-induced lung injury. AB - The effects of the beta-2-receptor agonist terbutaline on central hemodynamics, gas exchange, and platelet and leukocyte counts during 3 hr of endotoxin shock were studied. Ten sheep were anesthetized and ventilated without positive end expiratory pressure (PEEP). After 1 hr of stabilization (t = -30), five animals (Group T) received intravenous (i.v.) infusion of terbutaline, 20 micrograms/kg/hr for 3.5 hr, whereas the other five received no drug treatment and served as controls (Group C). Thirty min later (t = 0) all animals received Escherichia coli endotoxin 10 micrograms/kg by i.v. infusion over 15 min. The terbutaline infusion increased the heart rate (HR) initially by 30% and the cardiac index (CI) by 50%, whereas mean arterial pressure (MAP), pulmonary artery pressure (PAP), and gas exchange remained unchanged. Terbutaline pretreatment did not prevent the pulmonary hypertension that characteristically occurs after endotoxin injection, nor did it decrease the initial fall in platelet count, leukocyte count, arterial oxygen tension (PaO2), or respiratory compliance (t = 30). However, during the permeability phase (after 120-180 min), there was a significant improvement in MAP, PAP, respiratory compliance, PaO2, and arterial pH in the animals treated with terbutaline as compared with the control animals. Also, the wet weight to dry weight ratio of the lungs from animals receiving terbutaline was significantly lower than in controls. It was concluded that terbutaline does not influence the hypertension phase during endotoxin shock, but it may decrease pulmonary microvascular leakage during the permeability phase. PMID- 3048767 TI - Triacylglycerol output by the isolated perfused liver of endotoxin-treated rats. AB - Liver perfusion experiments were performed to further investigate the origin of hypertriacylglycerolemia associated with endotoxemia in rats. For this purpose male rats were treated with E. coli endotoxin in a dose of 0.5 mg/100 g b.w. intravenously, deprived of food (with free access to water), and used 18 h later for liver perfusion experiments. Livers were isolated and perfused for 4 h with a medium consisting of Krebs-Ringer bicarbonate buffer containing bovine serum albumin (4%, w/v), glucose, 5 mM, oleate, 0.7-0.8 mM, and aged human erythrocytes, 20% (v/v). The rates of triacylglycerol (TAG) output, oleate uptake, and ketogenesis were measured. Endotoxin treatment did not induce changes of the rate of either TAG output or ketogenesis in the perfused liver. The data give further support to the contention that hypertriacylglycerolemia associated with endotoxicosis is unlikely to originate in an overproduction of TAG by the liver. PMID- 3048768 TI - Beneficial effects of buprenorphine (a partial opiate agonist) in porcine Escherichia coli septicaemia: a comparison with naloxone. AB - The cardiovascular and metabolic responses to treatment with naloxone or buprenorphine (a partial opiate agonist) were investigated in a porcine model of septicaemia. Animals anaesthetised with alpha-chloralose were infused with live E. coli over two hours. They were then divided into three groups and received either naloxone (2 mg kg-1 + 1.5 mg kg-1 hr-1) or buprenorphine (0.3 mg kg-1) or an equivalent volume of normal saline. Treatment was started one hour after commencing the infusion, by which time a significant fall in cardiac index (CI), stroke index (SI), mean arterial pressure (MAP), and pH had occurred in all groups, together with a significant rise in mixed venous blood lactate and packed cell volume. Treatment with both naloxone and buprenorphine resulted in significant improvements in CI, pH, and base excess and in a fall in mixed venous lactate and packed cell volume. Although no significant effect on survival was seen at three hours after the start of treatment, buprenorphine may prove to be a suitable alternative to naloxone in the management of septic shock. PMID- 3048769 TI - Chronic hyperdynamic sepsis in the rat: I. Characterization of the animal model. AB - We describe a new rat model of chronic hyperdynamic sepsis. After control values for weight gain, and food and water intake of each animal were obtained over a 5 day period, male Sprague-Dawley rats weighing 370-425 g were anesthetized, catheterized to allow chronic cardiac-output measurements, and a sterile subcutaneous cavity was formed over the flank area. The animals were allowed a 3 4 day postoperative recovery period. Body weight, food and water intake, and cardiac output were measured daily. Frequent blood samples were withdrawn for bacterial cultures and white cell counts (WBC). On the third and, in some cases, the fourth postoperative day, the subcutaneous cavity was inoculated with 10(9) colony-forming units of Escherichia coli and Bacteroides fragilis. The resulting sepsis was characterized by loss of body weight in spite of normal food and water intake, increased cardiac output, increased WBC, intermittent bacteremia, decreased muscle mass, and decreased cross-sectional area of skeletal muscle myofibrils. Two levels of septic response emerged--moderate and severe. Based on the above-mentioned measurements, it was possible to categorize all long-term septic animals into these two groups. Both groups exhibited cardiac-output, body weight, and WBC data significantly different from sham controls. Repeated inoculations of the subcutaneous abscess initiated on the third postoperative day resulted in moderate sepsis with no long-term mortality, severe sepsis with 23% mortality over a 3-week period, or a 100% mortality within 4 days, depending on the virulence of the E. coli organisms used. The new model is ideally suited for pathophysiologic studies of sustained, hyperdynamic sepsis. PMID- 3048770 TI - The effects of chronic bacteremia on the metabolic response to exercise in the conscious rat. AB - Cardiac function was indirectly assessed in a rat model of chronic sepsis by measuring maximal oxygen uptake (VO2max) during exercise. A subcutaneous abscess cavity was created in rats by implanting a gauze sponge in the hindquarter area. Animals with sterile abscesses were compared with rats that had the abscess cavity infected with Escherichia coli and Bacteroides fragilis (gram-negative group) or these two organisms plus Staphylococcus aureus (gram-positive/negative group). VO2max was measured during exercise before and after the abscess cavity was infected. After a carotid artery cannula was placed, heart rate, mean arterial pressure, arterial blood gases, and acid-base status were measured in three groups of rats (sterile abscess, gram-negative, gram-positive/negative) during submaximal and maximal exercise. Results demonstrated that the infected rats were febrile and had elevated white blood cell counts and intermittent bacteremia, while rats with sterile abscesses did not. VO2max was similar in all groups of rats both before and after the abscess cavity was infected. The metabolic and hemodynamic variables measured during submaximal and maximal exercise in the different groups of rats were similar. These results do not support the contention that gram-positive and/or-negative bacteremia produce cardiac dysfunction during early sepsis. PMID- 3048771 TI - Naloxone potentiates epinephrine's pressor actions in endotoxemic rats. AB - Naloxone's pressor effects in shock may be mediated through antagonism of endogenous opioid inhibition of sympathoadrenal catecholaminergic systems. Since circulating catecholamine levels are not further elevated by naloxone in endotoxemic animals, it is possible that naloxone acts upon opiate receptors to enhance catecholamine actions at the receptor or postreceptor level. To investigate this hypothesis, we sought to determine whether naloxone treatment would augment the pressor actions of exogenous catecholamines (epinephrine) in normal and endotoxemic rats. Naloxone (3 mg/kg intravenous [i.v.] bolus followed by 3 mg/ml/hr infusion) significantly augmented the pressor response to 10, 20, and 50 micrograms/kg of i.v. epinephrine by 69%, 48% and 14%, respectively, in endotoxin (5 mg/kg, LD20, i.v.) -treated rats but not in normal rats. Likewise, the duration of the pressor response to epinephrine was significantly increased by naloxone. These findings suggest that the coadministration of naloxone and epinephrine may be of benefit in the treatment of shock. PMID- 3048772 TI - Canine peripheral vascular response to endotoxin shock at a constant cardiac output. AB - The peripheral vascular response to Escherichia coli endotoxin (1 mg/kg/i.v.) was measured for 2 hr in the pentobarbital anesthetized dog. Total venous return was collected and returned by a pump to the right atrium to maintain a constant cardiac output. Occlusion of the venous lines permitted estimation of venous compliance in the systems drained by the superior (SVC) and inferior vena cavae (IVC). After endotoxin administration, arterial pressure and total peripheral resistance rapidly dropped and remained low for 2 hr. IVC compliance was decreased at 10-30 min and SVC compliance at 10 min after endotoxin. The decrease in compliance is interpreted as venous dilation and probably venous pooling. The latter may account for a substantial portion of the total venous pooling reported in early endotoxin shock. After 30 min, compliance increased and by 60 min was equal to control. Left ventricular relaxation ability decreased as indicated by maximal negative dp/dt. Ibuprofen, 10 mg/kg, was administered at 120 min or earlier, depending on the state of the animal; rapid recovery of arterial pressure and ventricular function occurred without a significant change in venous compliance. PMID- 3048773 TI - Oxygen free radical activity during live E. coli septic shock in the dog. AB - Free radicals generated during purine catabolism or by activated granulocytes cause tissue injury by peroxidation of lipid membranes. In a canine model of sepsis initiated by intravenous live Escherichia coli, fluorescent products of lipid peroxidation (FP) were measured in serum. Four groups of five dogs infused with 10(9)E. coli/kg were analyzed--I: no further treatment; II: prior depletion of granulocytes with a cytotoxic antibody; III: pre-treatment with superoxide dismutase and catalase; and IV: resuscitation after bacterial infusion to maintain cardiac output greater than 80% of pre-bacteremic levels. In Groups I, II, and III, cardiac output fell to less than 50% of baseline within 1 hr and remained there throughout the study. FP in Groups I and II rose to greater than 200% of baseline (P less than .02 and less than .03). In Groups III and IV, FP did not rise significantly from baseline. The rise in serum FP and the prevention of this rise by-treatment with antioxidants indicate generation of oxygen radicals. Their presence had no effect on hemodynamic parameters. Granulocyte depletion did not alter appearance of FP; however, prevention of low cardiac output blocked FP formation. These data suggest that oxygen free radicals were generated by tissue ischemia, rather than by granulocytes, in this model of septic shock. PMID- 3048774 TI - Fatty acid analogue accumulation: a marker of myocyte viability in ischemic reperfused myocardium. AB - A 3-methyl substituted radioiodinated long chain fatty acid analogue was evaluated as an agent for the noninvasive detection of altered fatty acid uptake in reperfused, postischemic myocardium. This iodinated fatty acid analogue, 15 (para-iodophenyl)-3-methyl pentadecanoic acid, was given intravenously at 3 hours of reperfusion following 15 minutes (Group 1, n = 5 dogs) or 60 minutes (Group 2, n = 5 dogs) of left anterior descending coronary artery occlusion. Myocardial blood flow (MBF) was measured during occlusion and reperfusion with radiolabeled microspheres administered via the left atrium. Paired ultrasonic subendocardial crystals were placed in the ischemic perfusion bed to assess regional left ventricular systolic function at baseline, during ischemia and reperfusion. Electron microscopic analysis and staining with triphenyltetrazolium chloride (TTC) was performed. Groups 1 and 2 dogs had similar (p = NS) myocardial blood flows during occlusion. TTC positive 1 g endocardial segments from Group 1 (n = 98) and Group 2 (n = 71) had 37% greater fatty acid analogue activity (0.26 +/- 0.04 vs. 0.19 +/- 0.09 percent injected dose per gram; p less than 0.05) compared with TTC negative segments from Group 2 dogs (n = 37). When fatty acid analogue activity was related to near simultaneous reperfusion blood flow, this ratio was 27% greater (p less than 0.05) in TTC positive segments (0.38 +/- 0.1) compared with TTC negative (0.30 +/- 0.16) segments, and 9% greater than normal (0.35 +/- 0.09; p less than 0.05). While ischemic regions from both Groups 1 and 2 dogs became similarly dyskinetic during occlusion (systolic shortening, -11 +/- 6 vs. 11 +/- 2%; p = NS), TTC negative segments remained akinetic (= 1 +/- 7%) at 3 hours of reperfusion while TTC positive zones had recovered partial systolic function (8 +/- 22%). Electron microscopy confirmed the presence of reversible ultrastructural changes in TTC positive regions. A 60-minute occlusion, 3-hour reperfusion model adapted for in vivo single photon emission computed tomography showed a similar excess of 123I fatty acid activity over flow when compared to perfusion (as measured with 201Tl) in the ischemic border zone of 4/4 canine myocardial infarcts. We conclude that the accumulation of this non-beta-oxidized fatty acid analogue noninvasively identifies zones of discordance between fatty acid and flow distribution that are characteristic of ischemically "stunned" but viable myocardium. PMID- 3048775 TI - Xenon handling in the liver: red cell capacity effect. AB - Xenon, despite its lack of chemical reactivity, associates preferentially with red cells in blood. To characterize the effect of this and the nature of xenon tissue interaction in the liver, multiple indicator dilution studies were performed in the anesthetized normal dog through portal vein injection and hepatic vein collection of anaerobic blood samples. Two experimental runs were carried out in each animal, one at the prevailing hematocrit and the other at reduced hematocrit after bleeding and replacement with dextran. For comparison, the injection mixtures contained labeled red blood cells (a vascular reference), sucrose (an interstitial space reference), and labeled water (which freely enters liver cells), as well as labeled xenon. At the higher hematocrit, the labeled xenon curves generally rose earlier, peaked higher, and decayed more quickly than the labeled water curve; at the lower hematocrit, the xenon curve was delayed and diminished in magnitude in relation to the labeled water curves. Analysis of the curve shapes indicated that xenon, like labeled sucrose and water, underwent delayed wave flow-limited distribution. With knowledge of the red cell plasma partition coefficient (2.89 ml/ml), it was possible to both account for the change in form of the xenon curves with hematocrit and to use the data to estimate the liver cell tissue plasma xenon partition coefficient. Values averaged 1.93 ml/ml liver space, or 1.79 ml/g, and did not change significantly from first to second runs. Theoretical analysis indicated that flow cannot be estimated from xenon downslopes. PMID- 3048777 TI - Diagnostic utility of immunoassays for parathyrin in hyper- and hypocalcemic states. PMID- 3048776 TI - Hemodynamic and hormonal adaptations to beta-adrenoceptor blockade. A 24-hour study of acebutolol, atenolol, pindolol, and propranolol in hypertensive patients. AB - Comparison of the hemodynamic and hormonal effects of beta-adrenoceptor antagonists with different ancillary properties may help to clarify the antihypertensive mechanism of these drugs. Under strict basal conditions, the effects of acebutolol (400 mg b.i.d.), atenolol (100 mg b.i.d.), pindolol (10 mg b.i.d.), and propranolol (80 mg t.i.d.), were studied for the first 24 hours in 40 hypertensive patients. With pindolol, mean arterial pressure was reduced (p less than 0.05) 1 hour after administration, whereas the cardiac index and the systemic vascular resistance index did not change. With the other three drugs, the fall in mean arterial pressure was delayed 2-3 hours. With these drugs, the fall in mean arterial pressure was preceded by a rise in the resistance index, which compensated for the initial fall in cardiac index. With each drug, the decrements in mean arterial pressure were associated with parallel decrements in the resistance index, and percent changes in mean arterial pressure and the resistance index were always significantly (p less than 0.001) correlated. At the end of the 24-hour period, the four drugs shared an equal antihypertensive effect, which varied 14-17%. This was associated with a return of the cardiac index toward control values by acebutolol, atenolol, and propranolol treatment and a moderately increased cardiac index above pretreatment values (13%, p less than 0.01) with pindolol. The secondary rise in the cardiac index was inversely correlated (p less than 0.001) with the fall in mean arterial pressure with all four drugs. Plasma renin was maximally suppressed 2 hours after treatment, thus before any change in mean arterial pressure had occurred with acebutolol, atenolol, and propranolol. Pretreatment values of active renin and the reduction of mean arterial pressure 24 hours after administration were not correlated in any of the four groups. Despite the "vasodilator" action of the four drugs, plasma norepinephrine did not rise. Our data show that the main hemodynamic change that occurs at the time blood pressure falls after beta-adrenoceptor antagonism is vasodilation. Neither autoregulation of blood flow nor renin suppression can explain this vasodilator action. The absence of an increase in norepinephrine, despite vasodilation, suggests that beta-adrenoceptor antagonism interferes with sympathetic vasoconstrictor nerve activity. This effect may explain the vasodilator and antihypertensive potential of beta-adrenoceptor antagonists. PMID- 3048778 TI - Progesterone receptor: stability studies and correlation between steroid binding assay and enzyme immunoassay. AB - We evaluated and compared Abbott Laboratories' newly developed enzyme immunoassay (EIA) for measuring progesterone receptors (PgR) with that of DuPont's steroid binding assay (SBA). We also used both methods to study the stability of PgR under various conditions. THere were excellent correlations for all 59 cytosols compared (r = 0.94) and for the 44 cytosols containing PgR greater than 10 fmol per milligram of protein (r = 0.93), but the correlation for cytosols containing less than 10 fmol of PgR per milligram was poor. We found PgR to be more stable as assayed by EIA than by SBA. The biological half-lives of PgR at 30, 4, and -60 degrees C were approximately 3 h, 6 days, and 19 days, respectively. The effect of molybdate on PgR is complex. Its presence during tissue homogenization leads to analytical recovery of more PgR and may stabilize PgR during storage. Its presence during enzyme immunoassay is less critical. Unlike estrogen receptor, PgR is not protected by its ligand, R5020. PMID- 3048779 TI - Clinical variability of cyclosporine pharmacokinetics in adult and pediatric patients after renal, cardiac, hepatic, and bone-marrow transplants. AB - The most important limitation associated with the clinical use of cyclosporine is the narrow therapeutic range between its efficacy and toxicity. Effective treatment is further complicated by significant variation in intrapatient and interpatient pharmacokinetics of the drug. We describe a practical approach to pharmacokinetic analysis that does not interfere with the cyclosporine dosage regimen or with clinical management of the patient. To optimize therapy, we individualized patient management by using noncompartmental pharmacokinetic analysis. Mean residence time (MRT) and volume of distribution at steady-state were calculated from data on concentration vs time after dose. We applied this approach to 24 kidney, 12 heart, 8 bone-marrow, 7 liver, and 5 pancreas transplants. Individualized requirements for cyclosporine dose and dosage interval can be predicted from these parameters. MRT is the most useful pharmacokinetic parameter, because it allows prediction of the optimal dosage interval. PMID- 3048780 TI - High-performance liquid affinity chromatography: rapid immunoanalysis of transferrin in serum. AB - We describe a new method for quantitatively measuring substances of clinical interest by high-performance liquid affinity chromatography (HPLAC). As a model system we selected analysis for transferrin in human serum with immobilized antibodies in a high-performance liquid chromatographic system. SelectiSpher-10 Activated Tresyl columns (5 or 10 x 0.5 cm) were used for in situ coupling of polyclonal antibodies to transferrin. The amount of transferrin eluted was determined by integrating the eluted peak at 280 nm. The whole analytical procedure--including injection of sample, washing, elution, and analysis of data- takes only 7 min. We characterized the HPLAC system for analysis of transferrin in several ways: intra-assay CV approximately 3%; inter-assay CV 2-9%; linear response up to 1 mg/mL column volume; detection limit approximately 3 micrograms; analytical recovery 98% +/- 2%; purity of eluted sample greater than 95% (SDS PAGE). The HPLAC method was compared with "rocket" immunoelectrophoresis, a commonly used method of analysis for transferrin, and there was excellent correlation between the two methods (r = 0.96, n = 60). Benefits of this HPLAC technique include high precision, rapid analysis, and simplified sample handling. PMID- 3048781 TI - Evaluation of an enzyme immunoassay kit for estrogen receptor measurement--report II. AB - We present evidence to show that monoclonal antibodies to estrogen receptors (ER) in solid phase recognize the secondary estrogen binding sites with moderate to low affinity for estradiol (E2). An excellent quantitative agreement was found in five cytosols between the ER values obtained by the enzyme immunoassay (ER-EIA) and the amount of secondary estrogen binding sites measured by the assay involving dextran-coated charcoal (Clin Chem 1986;32:1496). The immunoreactive protein recognized by the antibody-coated beads, when allowed to react with ER(+) cytosols, is shown to bind [3H]estradiol only when the ligand concentration exceeds 8 nmol/L. Further biochemical and functional characterization of the immunoreactive protein is required to establish similarities/dissimilarities between this protein, high-affinity Type I ER sites, and the secondary sites such as Type II sites. PMID- 3048782 TI - Measurement of plasma phenytoin by EMIT in the Monarch centrifugal analyzer: studies on the elimination of within-rotor drift. AB - We investigated the measurement of phenytoin in plasma by the EMIT method, in the Monarch centrifugal analyzer. When we used the standard protocol supplied by Instrumentation Laboratory, significant drift was observed across a full rotor loaded with 31 replicates of a single specimen. Although the cause remains obscure, the drift was eliminated by using a load-spin-reload-spin procedure. This makes it possible to analyze large batches of samples without excessive use of standards and reagents. PMID- 3048783 TI - A laboratory and clinical evaluation of an immunochemiluminometric assay of thyrotropin in serum. AB - We evaluated an immunochemiluminometric assay for human thyrotropin. A chemiluminescent acridinium ester is used as a label, with magnetic-particle separation. The lower limit of detection of the assay (mean + 3 SD of the zero standard) was 0.07 milli-int. unit/L, with a working range of 0.5 to greater than 60.0 milli-int. units/L. Assay accuracy was good as judged from analytical recovery, analysis of external quality-assessment samples, and comparison with an enzyme-amplified immunoassay. There were no significant interferences or cross reactivities. Twenty-four samples assayed showed aggregation of the magnetic particles. On re-assay, four of these samples showed a significant increase in the measured TSH by the luminescence assay. Assay time for 60 tubes was approximately 3.5 h with the use of a semi-automated luminometer. The reference interval, determined from data on 144 healthy euthyroid subjects, was 0.3-4.0 milli-int. units/L. Sixteen of 19 thyrotoxic patients showed clearly suppressed concentrations of thyrotropin in serum. PMID- 3048784 TI - Multicenter evaluation of a specific pancreatic isoamylase assay based on a double monoclonal-antibody technique. AB - Eleven evaluators from nine laboratories in five countries evaluated a new immunoinhibition method for pancreatic isoamylase determination that is as simple to perform as that for total amylase. The precision at low and intermediate activity concentrations was superior, and at high concentrations it equalled that of the wheat-germ inhibitor method. The test was linear to approximately 2000 U/L, depending on the instrumentation used. The percentage salivary isoamylase activities remaining in specimens after reaction with two monoclonal antibodies ranged from 2 to 4.4%. Comparative studies showed good correlation with the wheat germ inhibitor (r greater than 0.978) and electrophoresis methods (r = 0.920). Hemolysis, lipemia, and bilirubinemia have no effect on results. Interlaboratory studies demonstrated excellent transferability of the method, if instruments are calibrated with the same calibrator. Reference intervals for pancreatic isoamylase are 13 to 64 U/L (25 degrees C), 13 to 83 U/L (30 degrees C), and 17 to 115 U/L (37 degrees C). A clinical evaluation of patients with acute pancreatitis showed that pancreatic isoamylase has a greater clinical sensitivity than total amylase. PMID- 3048785 TI - An immunoenzymatic method for pancreatic oncofetal antigen automated in the Boehringer ES 600. PMID- 3048786 TI - Possible interference by amiodarone and desethylamiodarone in thyroxin assays. PMID- 3048787 TI - "Stratus" fluorometric enzyme immunoassay system evaluated for determination of free thyroxin. PMID- 3048788 TI - Preliminary assessment of the "Magic Lite" CK-MB immunoassay. PMID- 3048789 TI - Effect of immunosuppressive treatment on aminotransferase activities in serum of renal-transplant recipients. PMID- 3048790 TI - Immunometric assays may underestimate thyrotropin concentrations in sera from infants with congenital hypothyroidism. PMID- 3048791 TI - Facial growth in the rabbit after autologous grafting in unilateral clefts. AB - In each of 39 rabbits, a left-sided cleft of the lip and alveolar part of the maxilla were made by removal of the premaxillomaxillary suture. In 5 animals, the cleft was left unrepaired. In 12 animals, an autologous graft from the mandibular symphysis was introduced, and in 12 animals autologous rib-growth cartilage was used for grafting. Lateral photographs of adult skulls in a standardized position were made, and a computerized craniometric analysis of the position of 13 anatomic landmarks in an orthogonal coordinate system was performed. The data from the grafted skulls (12) were compared with those from the control series (20) and subjected to a multivariate analysis, according to Hotelling. Comparison of the grafted adult skulls with 20 unoperated controls and the animals with unrepaired artificial clefts revealed that introduction of rib-growth cartilage leads to an improvement with respect to the growth of the facial skeleton. The initial orientation of the graft in the cleft appeared to be important for its integration in the jaw. PMID- 3048792 TI - Effect of a new somatostatin analogue SMS 201-995 (Sandostatin) on the renin aldosterone axis. AB - The effects of a new somatostatin analogue SMS 201-995 (H-D-Phe-Cys-Phe-D-Trp-Lys Thr-Cys-Thr(ol), Sandostatin) on the orthostatic stimulation of plasma renin activity (PRA) following head-up tilting and on angiotensin II (Ang-II) induced aldosterone (PA) release were studied under placebo controlled conditions in separate groups of healthy volunteers consisting of six and 10 subjects, respectively. Head-up tilting (by 60 degrees) produced the characteristic increases in PRA and PA. Administration of SMS 201-995 significantly (P less than 0.05) inhibited this PRA elevation from 30 min on. Throughout the study period, PA levels were not consistently altered by this analogue. Furthermore, SMS 201 995 failed to inhibit the stimulation of PA secretion induced by exogenous angiotensin-II (2-10 ng/kg/min). Results presented here are at variance with data collected with natural somatostatin showing an inhibitory effect on stimulated PA. This discrepancy can be explained by the recently described absence of SMS 201-995 binding sites in primate adrenal cortex and in human aldosteronomas. PMID- 3048793 TI - Basal and stimulated insulin levels rise with advancing puberty. AB - We studied the effect of pubertal development on insulin secretion. Intravenous glucose tolerance tests were performed on 47 islet-cell antibody negative siblings of diabetic children and on 16 normal adults. Puberty was staged using Tanner's criteria and subjects were grouped as follows: I, stage 1 (n = 16); II, stages 2 and 3 (n = 15); III, stages 4 and 5 (n = 16); IV, adults (n = 16). Fasting insulin increased with advancing puberty (p = 0.59, P less than 0.001). The stimulated insulin response also rose with increasing pubertal development: for the 0-10 min insulin area, p = 0.46, P less than 0.001 and for the 10-60 min area, p = 0.68, P less than 0.001. There was a low positive correlation between insulin and age to 16 years but multiple regression analysis showed that this could be accounted for by puberty alone. Indeed prepubertal children and adults did not differ. There was no correlation between glucose (fasting and 0-60 min area) and puberty or age. These findings suggest that insulin resistance increases during puberty, and this may contribute to the frequency of presentation or worsening control of insulin dependent diabetes at this time. PMID- 3048794 TI - The mechanism of the adolescent growth spurt induced by low dose pulsatile GnRH treatment. AB - We have used GnRH administered in a pulsatile fashion to treat 26 patients (12M:14F) with delayed puberty. Treatment was for a mean of 1.05 years (range, 0.3-1.6). Mean age at the onset of treatment was 16.4 years in the girls (range, 12.7-28.2) and 15.8 years in the boys (range, 13.8-17.8). At different stages of sexual maturation, overnight serum samples for growth hormone (GH) were taken at 15 min intervals between 2000 h and 0600 h. The girls had a peak growth velocity which occurred between breast stages 2 and stage 3 (B2/3). GH secretion (both sum of the GH peaks and area under the GH pulse) increased at B2 and reached a peak at B3. Growth acceleration in the boys started at the attainment of a 9-10 ml testicular volume and reached a peak at 11-15 ml. The boys demonstrated an initial fall in GH secretion with the onset of treatment until the attainment of 9-10 ml testicular volume; peak GH secretion occurred at the attainment of 11-12 ml testicular volume. There was no change in GH pulse frequency during treatment in either sex. These observations and the maintenance of the normal relationship of the growth spurts to the appearance of secondary sexual characteristics are relevant to the mechanisms and timing of the adolescent growth spurts in normal girls and boys. PMID- 3048796 TI - Luteinizing hormone response to domperidone in hyperprolactinaemic women with pituitary microadenomas. AB - Administration of a dopamine (DA) antagonist, domperidone, increased circulating levels of LH in hyperprolactinaemic-amenorrhoeic women with pituitary microadenomas but not in normal women in the early follicular phase of the menstrual cycle. This drug does not readily cross the blood-brain barrier and therefore the site of action of domperidone could be the pituitary gland or the median eminence. As the simultaneous administration of GnRH and domperidone did not increase the GnRH-induced LH release in hyperprolactinaemic women, the site of action of domperidone is likely to be the median eminence. PMID- 3048795 TI - Dysgerminoma in 45,X Turner syndrome: report of a case. AB - Here we report the fourth case of dysgerminoma in a patient with the syndrome of gonadal dysgenesis and 45,X karyotype. Typical Turner's syndrome features were unusually associated with breast development, menarche and secondary amenorrhoea. Exaggerated basal and GnRH stimulated gonadotrophin and low oestradiol levels were typical of post-pubertal Turner's syndrome. Detailed (standard) chromosome and banding analysis excluded the presence of Y chromosome material. This case suggests that the presence of a Y chromosome is not necessary for abnormal differentiation of germ cells and the occurrence of a gonadoblastoma. PMID- 3048797 TI - Peritoneal lavage as an aid in the diagnosis of acute peritonitis of non traumatic origin. PMID- 3048798 TI - Metabolism of ethanol and its consequences for the liver and gastrointestinal tract. PMID- 3048799 TI - Enteric nerves and function of intestinal mucosa. Physiological and pathophysiological aspects. PMID- 3048800 TI - Coeliac sprue: a centennial overview 1888-1988. PMID- 3048802 TI - Amniotic fluid folate, vitamin B12 and transcobalamins in neural tube defects. AB - Levels of folate, vitamin B12, the vitamin B12 binding proteins, apotranscobalamin I, II and III (TC I, II and III) and the unsaturated vitamin B12 binding capacity (UBBC) were measured in mid-trimester amniotic fluids from normal pregnancies, and from those where the fetus had open spina bifida, anencephaly or omphalocoele, and where the fetus was normal but the mother had had a previous neural tube defect pregnancy. At 15-19 weeks' gestation, vitamin B12 levels were low in the fluids of all the types of abnormal fetuses, and also of normal fetuses where there had been a previous NTD sib. In contradistinction, TC I, II and III and UBBC levels were generally abnormally high in all these groups. Low vitamin B12 levels in the face of high carrier protein levels suggest deranged vitamin B12 production or transport. Since these abnormalities are present in fluids from normal sibs of NTD individuals as well as from those with midline lesions, an inherited defect is implied. We propose that at least part of the genetic predisposition to NTD, and possibly other midline defects, could reside in an abnormality connected with vitamin B12 production, transport or metabolism, and a mechanism is suggested. PMID- 3048801 TI - Motor activity of the distal ileum and ileocecal sphincter and its relation to the regions's function. PMID- 3048803 TI - An analysis of myeloma plasma cell phenotype using antibodies defined at the IIIrd International Workshop on Human Leucocyte Differentiation Antigens. AB - Fresh bone marrow from 43 cases of myeloma and three cases of plasma cell leukaemia has been phenotyped both by indirect immune-rosetting and, on fixed cytospin preparations, by indirect immunofluorescence. Both clustered and unclustered B cell associated antibodies from the IIIrd International Workshop on Human Leucocyte Differentiation Antigens were used. The results confirm the lack of many pan-B antigens on the surface of myeloma plasma cells, i.e. CD19-23, 37, 39, w40. Strong surface reactivity is seen with CD38 antibodies and with one CD24 antibody (HB8). Weak reactions are sometimes obtained with CD9, 10 and 45R. On cytospin preparations CD37, 39 and w40 are sometimes weakly positive, and anti rough endoplasmic reticulum antibodies are always strongly positive. Specific and surface-reacting antiplasma cell antibodies are still lacking. PMID- 3048804 TI - One half of the CD11b+ human peripheral blood T lymphocytes coexpresses the S-100 protein. AB - The expression of the CD11b antigen and the presence of the S-100 (and, specifically, its beta subunit) protein within the T4- subpopulation of normal human peripheral blood lymphocytes were investigated by panning techniques, immunofluorescence analysis and immunoelectronmicroscopy. Both antigens are known to be absent in the T4+ lymphocytes. However, CD11b+ T lymphocytes represented about 30% of the T4- population; a part of them (over 1/3) belonged to and completely filled up the T4- T8- subpopulation, whereas the remaining part (almost 2/3) shared the T8 positivity. Interestingly, S-100+ T lymphocytes, which always were CD11b+ too, represented about one half of the CD11b+ T cells, but were excluded from the T4- T8- CD11b+ subpopulation, whereas they represented up to 80% of the T4- T8+ CD11b+ subset. Such findings demonstrate that the S-100+ T lymphocytes are exclusively restricted to a discrete T cell compartment which shows the T8+ CD11b+ immunophenotype. Since such T8+ CD11b+ cells had been shown to possess suppressive capabilities, we herein propose that S-100+ lymphocytes might to some extent modulate the immune responses. However, the exact functional significance of the S-100 protein still remains unknown. PMID- 3048805 TI - Nonimmune human cells can express MHC class II antigens in the absence of invariant chain--an immunohistological study on normal and chronically inflamed small intestine. AB - The expression of HLA-DR, HLA-DP and HLA-DQ antigens and of the associated invariant chain (Ii) was studied immunohistologically in sessile cells of normal ileum and ileum affected by Crohn's disease which was taken as a model for chronic inflammation. Corresponding to the local inflammatory cell density, a considerable neo-expression of MHC class II antigens and Ii was observed in epithelial cells, vascular endothelial cells, and Schwann cells of the enteric nerve plexus. While HLA-DP and HLA-DQ antigens were undetectable in sessile cells of normal ileum, class II antigens expression in ileitis followed the order HLA DR greater than or equal to HLA-DP greater than or equal to HLA-DQ. In various cell types a differential expression of Ii and class II antigens was noted. In normal crypt enterocytes and in arterial endothelial cells of inflamed ileum, Ii was found in the absence of class II antigens. On the other hand, most Schwann cells and internodal nerve strands of the submucous and myenteric plexus exhibited HLA-DR (-DP, -DQ) antigens in the absence of detectable Ii. Moreover, HLA-DR+ endothelial cells in normal tissue specimens were Ii-, and in inflamed intestine HLA-DR expression of venous/venular and capillary endothelium greatly exceeded Ii expression. The observation of both Ii+/HLA-DR- and Ii-/HLA-DR+ (HLA DP+, HLA-DQ+) cells questions the previously assumed close association of Ii and class II antigen expression. PMID- 3048806 TI - Differential isotype recognition of two centromere associated polypeptides by immunoblotting in connective tissue disease. AB - On investigating the immunoblotting profile of 65 systemic sclerosis patients, a 140 kD polypeptide was recognised by sera from 16, when immunoblotted against a nuclear-enriched K562 cell sonicate. All 16 sera contained anticentromere antibodies (ACA) detected by immunofluorescence (IF) and 15 of 16 also recognized a 19 kD polypeptide on immunoblotting. Two ACA positive sera failed to recognize the 140 kD polypeptide but one of these recognized the 19 kD polypeptide. The 140 kD polypeptide identified a group with more limited skin involvement (P less than 0.05) and all 16 had Raynaud's phenomenon. The sera from three of 100 systemic lupus erythematosus (SLE) patients also recognized both polypeptides. On investigating the isotype specificity, the 140 kD polypeptide was strongly detected by an IgM autoantibody and the 19 kD polypeptide by an IgG autoantibody. PMID- 3048807 TI - Proteins responsible for anticentromere activity found in the sera of patients with CREST-associated Raynaud's phenomenon. AB - Anticentromere antibodies (ACA) present in a high percentage of patients with complete or incomplete CREST scleroderma, and which are presently used in the diagnosis of this disease, also appear in some primary Raynaud's phenomenon patients. Three principal centromeric antigens, CENP-A, CENP-B and CENP-C, have been described as reacting with the sera of these individuals. We attempt to determine whether or not a correlation between the presence of ACA and serum reactivity against one or more of these peptides could be established, and have observed that CENP-A, but not CENP-B or CENP-C, is specifically recognized by all patients sera tested. The fact that this reactivity is clearly detectable at very high serum dilutions, thus eliminating other non-specific interference, suggests that anti-CENP-A activity might be useful in the diagnosis of patients with CREST associated Raynaud's phenomenon. PMID- 3048808 TI - Decreased expression of CD7 occurs in rheumatoid arthritis. AB - To evaluate the possibility of antigenic modulation in vivo, we studied the expression of the CD7 antigen on the surface of peripheral blood and intrasynovial lymphocytes from patients with rheumatoid arthritis (RA). This disease is noted for features predicting changes in CD7: (1) increased expression related to activation during an immune response through possible contact with 'disease associated antigens' and (2) decreased expression associated with lymphocytes moving through tissue. We found a highly significant decrease of CD7 on RA T cells in vivo (P less than 10(-4] that was not related to disease activity, drug ingestion, or abnormalities in the ability of RA T cells to express or modulate the antigen in vitro. Similar decreased expression was observed on many intrasynovial T cells that were nevertheless activated as measured by expression of activation markers such as Act I (a late lymphocyte activation antigen) and Act II (the transferrin receptor). Decreased expression of CD7 occurs naturally in vivo in RA; this observation may have future significance in better understanding the immunopathogenesis of this disease. PMID- 3048809 TI - Two calcium-binding proteins associated with specific stages of myeloid cell differentiation are expressed by subsets of macrophages in inflammatory tissues. AB - Using a monoclonal antibody to macrophage migration inhibition factor (MIF), two proteins were isolated from supernatants of Concanavalin A-stimulated human peripheral blood mononuclear cells which seem to have complexed to a third component carrying the MIF activity. They are therefore designated MIF-related proteins or MRP-8 and MRP-14 according to their apparent molecular weights. Partial amino acid sequences have been determined and their cDNA have been cloned and expressed in Escherichia coli. Both are calcium-binding proteins and MRP-8 seems to be largely homologous to the cystic fibrosis antigen (Dorin et al., 1987). Antisera were raised in the rabbit against the recombinant proteins and their expression in cells and tissues studied using immunohistological techniques. The proteins are only found in blood granulocytes and monocytes. In culture the number of positive monocytes sharply increased and then declined with time, suggesting that their expression is associated with early stages of monocyte/macrophage differentiation and absent from resident macrophages in all tested tissues. In acute inflammatory reactions, e.g. gingivitis, MRP-8 is never seen in the tissue, whereas MRP-14 is expressed by intravascular monocytes and perivascular macrophages. In contrast, in chronic inflammation, e.g. rheumatoid arthritis, MRP-8 is also expressed by macrophages in the tissue. From this it is concluded that MRP-8 and MRP-14 are expressed sequentially at defined stages of monocyte/macrophage differentiation and that dysregulation of this process in chronic inflammation is mirrored by the presence of MRP-8-positive macrophages in the tissue. PMID- 3048810 TI - A monoclonal DNA-binding autoantibody causes a deterioration in renal function in MRL mice with lupus disease. AB - Two DNA-binding monoclonal antibodies (MoAb) derived from lupus mice were examined for their effects on kidney function. Antibody IV-228, reactive with single-stranded DNA (ssDNA) but not double-stranded DNA (dsDNA) significantly altered kidney function in some MRL/Mp-lpr/lpr (MRL/lpr) mice with lupus glomerulonephritis. Antibody II-410, preferentially reactive with dsDNA, did not modify kidney function in MRL/lpr mice. Neither antibody affected normal healthy MRL/Mp- +/+ (MRL/n) mice. Not all MRL/lpr mice were equally affected by IV-228 and in some animals with clinical disease it was without effect even when high circulating antibody levels were maintained by five sequential daily injections. Its affects upon kidney function appeared to be related to its ability to localize in vivo in glomeruli. It is assumed that epitopes on trapped immune complexes are immunologically available to circulating antibodies, and in those animals where injected antibody is not captured this is because the antigen epitope is either absent or is masked by the animals' own auto-antibodies. We conclude that antibodies which bind to ssDNA contribute to lupus nephritis; that individual mice make DNA-binding antibodies of different fine specificity; and that kidney disease is not always due to the same balance of antibodies in members of a genetically homogeneous stock. PMID- 3048811 TI - IgD in nasopharyngeal secretions and tonsils from otitis-prone children. AB - Quantification of IgD was performed by ELISA in 180 plasma samples and 83 nasopharyngeal secretions (NPS) from children aged 2-162 months with varying degrees of recurrent otitis media. Furthermore, in 24 of the children the density of the IgD-immunocytes (IgD-cells) was calculated in immunoenzyme-stained cross sections of their nasopharyngeal tonsils (NPT). Owing to a considerable variation of the IgD cell density throughout the NPT, a semi-quantitative counting system was applied. An irregular distribution of plasma IgD was observed during childhood and maximum levels were found between 48 and 72 months of age. However, an even distribution of plasma IgD was found among the four groups of children investigated. Based on calculations of the transudation of albumin from plasma to the NPS the amount of locally produced IgD in NPS (NPS-IgD (local] was estimated to 88% (range 41-99%). Significantly higher levels of NPS-IgD (local) were found in otitis-prone children than in the other groups. Moreover, a positive correlation was calculated between levels of NPS-IgD (local) and NPT-IgD cell density, indicating that NPT, being the local lymphoepithelial tissue, also functions as an important source of NPS-IgD. NPS-IgD was not found to be associated with secretory component, indicating a passive transfer of IgD through the mucosal membranes. Our results support the hypothesis of an association between the occurrence of IgD in the mucosa and secretions of the upper respiratory tract with localized inflammatory events. PMID- 3048813 TI - Functional and phenotypical studies of the Leu-4 (CD3)+, Leu-1 (CD5)- T lymphocyte. AB - A small T cell subpopulation expressing the phenotype Leu-5(CD2)+, Leu-4(CD3)+, Leu-1 (CD5)- can be found in peripheral blood and bone marrow of normal individuals. When these cells were sorted out by three colour immunofluorescence cell sorting and tested in limiting dilution assays, they were found to have lower frequencies of proliferating (9.0 +/- 5.6 times, n = 7) and of IL-2 producing cells (11.5 +/- 5.0 times n = 5), and a higher frequency of cytotoxic cells (3.1 +/- 2.6 times, n = 2) than T lymphocytes expressing the three markers. In peripheral blood lymphocytes, 1/3 of the CD3+, CD5- cells were positive for Leu-2a (CD8) while virtually all were negative for Leu-3a (CD4). Four colour flow cytometric analysis revealed a small subset of T cells positive for CD3 and negative for CD5, CD4 and CD8. Approximately 75% of the CD3+, CD5- cells were negative for Leu-7 and CD16 simultaneously. These results shed a light on the phenotype of T cells that escape killing by CD5 and complement in T cell depleted bone marrow and may explain why fewer residual T cells in the depleted marrow are detected by limiting dilution assays than by flow cytometric analysis. PMID- 3048812 TI - The effect of random blood transfusions on immunoglobulin production by peripheral blood mononuclear cells from uraemic patients. AB - The effect of blood transfusion on humoral immunity in chronic renal failure was studied by examining immunoglobulin production in vitro, in patients awaiting renal transplantation. Pokeweed mitogen (PWM) induced IgG plaque formation was normal in non-transfused uraemic patients while both spontaneous and Staphylococcus aureus Cowan I (SAC) induced immunoglobulin production were reduced. Five to ten units of third party blood transfusion reduced PWM-driven B cell differentiation, but had no effect on SAC-induced plaque formation, while spontaneous production of immunoglobulin was either enhanced or unaffected. As it is known that the response to SAC is less affected by suppressor T cell activity than that to PWM, these differences in the inhibitory effects of blood transfusion on B cell differentiation are further evidence that transfusion may act by increasing suppressor T cell activity. PMID- 3048814 TI - HLA-DR expression in the vascular lesion and circulating T lymphocytes of patients with giant cell arteritis. AB - Giant cell arteritis (GCA) is a vascular inflammatory disease characterized by accumulations of T lymphocytes and macrophages in the arterial wall. In order to characterize the immune response in GCA, we have analysed temporal artery biopsies using a double-staining immunofluorescence method and studied T lymphocytes in peripheral blood with a fluorescence-activated cell sorter (FACS). HLA-DR was expressed on 28% (range 16-42%) of all T lymphocytes in the wall of the inflamed temporal artery, but only on average on 6% of peripheral blood T lymphocytes, indicating a high degree of local T cell activation in the inflammatory lesion. The proportions of T lymphocytes, T helper/inducer and T suppressor/cytotoxic cells in the blood of GCA patients before treatment with prednisolone did not deviate from those in normal individuals, but there was a minor increase in T helper cells after initiation of steroid therapy. The number of T helper cells expressing HLA-DR antigen and IL-2 receptor was not altered after 6-10 days of treatment with prednisolone. We found no evidence of HLA-DR expression by arterial smooth muscle cells in the GCA lesions, suggesting that these cells do not serve as antigen-presenting cells in GCA. PMID- 3048816 TI - Structure and function of the skin. Are there differences between black and white skin? AB - In concluding my remarks on structure and function of the skin, I wish to make one additional observation. Most dermatologists have acknowledged the role of the skin and its ability to protect the human organism from invasion by pathogenic organisms. There is good scientific evidence that skin plays an important protective role. Although an intact epidermis is of utmost importance as a protective unit, the contribution of the "acid mantle" is of considerable magnitude. The acidity or alkalinity and stability of this mantle is contributed to, in large measure, by the presence of eccrine gland secretions and sebaceous gland secretions. It is then not logical to assume that if black skin contains larger numbers of large and overly active adnexal glands, it would have a most effective mechanism for control of bacterial, viral, and other infections? That this is not the situation becomes apparent in the discussions that follow on cutaneous infections in black skin. PMID- 3048815 TI - Induction of anti-nuclear antibodies in mice orally exposed to cadmium at low concentrations. AB - Anti-nuclear antibodies (ANA) in serum were detected in male ICR mice fed drinking water containing 3, 30 and 300 ppm Cd as CdCl2 for 10 weeks. In response to Cd exposure, ICR mice developed ANA of the IgG class giving nuclear patterns moderately stained by immunofluorescence or immunoenzyme method. Positive immunofluorescence staining of ANA was obtained in 50, 89 and 90% of ICR mice exposed to 3, 30 and 300 ppm Cd, respectively. Their spleen cells also showed an enhancement of antibody forming response to sheep red blood cells (SRBC) without the SRBC priming. When mice were primed with SRBC after exposure to Cd, however, a significant suppression of the antibody forming response was observed in mice fed 300 ppm Cd but not in those fed 3 ppm Cd. No significant differences in delayed-type hypersensitivity reaction to SRBC were observed between Cd-fed and control animals. Inbred BALB/c mice were less susceptible to the induction of ANA by Cd, as induced only in 300 ppm Cd-fed mice. Thus environmental exposure to Cd can induce ANA in ICR mice with a high susceptibility, presumably accompanied with a non-specific stimulation of antibody formation. PMID- 3048818 TI - Contact dermatitis in blacks. AB - Black skin is characterized by structural and functional differences such as increased stratum corneum cohesion, melanin content, and stratum corneum layers. These differences seem to make black skin difficult for irritants and light to penetrate, thus explaining the common opinion that skin in blacks is harder and develops contact dermatitis less frequently. The paucity of interpretable epidemiologic data and of clinical and experimental studies does not permit confirmation of this hypothesis, and the few data available are controversial. This article describes the main physiologic differences between black and white barrier function and reviews the literature on irritation, sensitization, and transcutaneous penetration. We found that the data are still too incomplete to generalize on the resistance, or lack thereof, of black skin (versus white skin) to chemical irritation, sensitization, and penetration. PMID- 3048817 TI - Cutaneous reaction patterns in blacks. AB - The study of black skin continues to be an important part of the field of dermatology. Black skin is unique and has as the result demonstrable complexities that are often inherent in diagnosing its disorders accurately. Even when the more common dermatoses are encountered on black skin, a keen diagnostic acumen is required in order to adjust for the presence of pigmentation and still arrive at the correct diagnosis. This pigmentation, coupled with the common tendency of black skin to present peculiar patterns of reaction to even the most common dermatoses, contributes to the confusion that often results when such a patient presents for dermatologic consultation. Some dermatoses that are of concern to the patient and physician alike may be nothing more than part of the patient's "blackness"; those conditions that are merely normal variants. Then, too, it is important to be aware of the pharmacologic effects of dermatologic therapy on black skin in order to minimize the undesirable side effects. It is hoped that the reader has, through this brief overview, gained a greater respect for and understanding of the uniqueness that defines black skin. PMID- 3048820 TI - Cosmetic surgery of black skin. AB - Black patients who desire cosmetic surgery have not had many treatment options or services available to them. At this time, the requests for specific cosmetic surgical procedures by black patients have far exceeded the number of cosmetic surgeons able to meet their needs. A surgeon who calls himself or herself a cosmetic surgeon can no longer limit professional services to white patients only. The patient selection process must be color-blind, for only then will the patient perceive that his or her doctor truly has a sense of ethnic fair play. PMID- 3048819 TI - Photosensitivity reactions in black skin. AB - The findings and discussions in this article clearly point out the need for a better understanding not only of the general field of photobiology and photosensitivity reactions but in particular of the role of melanin as well as the genetic bases of these diseases that have not been clearly defined to date. In other words, these discussions clearly demonstrate the need for comprehensive research into the area of photosensitivity in black skin. The fact that black individuals tend to avoid tanning as well as to avoid prolonged exposure to sunlight because of the tremendous heat load that it induces, with consequent discomfort, undoubtedly has played a role in the lower incidence of photosensitivity reactions diagnosed to date. However, this has changed, and the incidence of adverse effects has been increasing in recent years since sunning became fashionable. PMID- 3048821 TI - Disorders of the hair and scalp in blacks. AB - The hair follicle and shaft in blacks have unique configurations. The occurrence of certain disorders such as hot-comb and traction alopecia, proximal trichorrhexis nodosa, trichonodoses, and pseudofolliculitis predominantly among blacks is related to the morphology as well as methods of caring for black hair. In addition, there are several disorders of unknown etiology such as dissecting cellulitis that also occur primarily in blacks. Familiarity with all of the structural differences that distinguish black hair and the disorders that affect the black patient's hair and scalp will enable dermatologists to provide proper diagnoses, treatment, and counselling for their black patients. PMID- 3048822 TI - Skin cancer in blacks in the United States. AB - Skin cancer is rare in blacks compared with whites in the United States. The most common form is squamous-cell carcinoma, not basal-cell carcinoma, as it is in whites. Sunlight does not appear to be an important etiologic factor in skin cancer in blacks, as most lesions occur on covered areas. Malignant melanoma is low in frequency but commonly affects acral areas and has a poor prognosis. Mycosis fungoides and dermatofibrosarcoma protuberans appear to have a high frequency among skin cancers. Squamous-cell carcinoma, malignant melanoma, and mycosis fungoides have a relatively high mortality rate in blacks. Bowen's disease and Kaposi's sarcoma occur in blacks but are rare. As there is a high frequency of squamous-cell carcinoma of the skin in blacks, prevention and early detection should benefit the patient. Considering the difficulties encountered in applying epidemiologic methods to skin cancer on a national scale, etiologic studies should be conducted in carefully selected areas. Future investigations of skin cancer in blacks should include an examination of risk factors such as burns, trauma, and diet and familial and immunologic aspects as well. PMID- 3048824 TI - Keloids. AB - Keloids are benign fibrous growths that result from an abnormal connective tissue response in certain predisposed individuals. Blacks form keloids more often than whites; however, the reason for this racial difference is not known. Trauma, foreign-body reactions, infections, and endocrine dysfunction have all been proposed as precipitating factors. Keloids are found most commonly on the ear lobes, shoulders, upper back, and midchest. They extend past the area of trauma and once present tend to remain stable. Although sometimes pruritic, painful, or tender, they are usually asymptomatic. Histologically, keloids are characterized by thick collagen bundles, abundant mucinous ground substance, few fibroblasts, and few if any foreign-body reactions. Although there have been many therapeutic modalities, most have had limited success. The most commonly used therapeutic approach is a combination of cryotherapy, intralesional steroid injections, surgical excision, and pressure devices. PMID- 3048823 TI - Pseudofolliculitis barbae and related disorders. AB - Pseudofolliculitis barbae, although not a serious medical problem, is certainly a distressing one for the affected patient. Its pathogenesis lies in an ingrown hair arising from the curved hair and follicle common in black men and women. Improper shaving techniques cause ingrown hairs through both transfollicular and extrafollicular mechanisms. Various treatment modalities exist, but there is no cure. Treatment must be individualized, as not all regimens will work for each patient. With diligence, pseudofolliculitis barbae can in many instances be controlled. Dermatitis papillaris capillitii is related to pseudofolliculitis barbae because its pathogenesis also lies in a curved hair and follicle. The treatment differs, however. Mild to moderately severe cases can be kept under good control with intralesional injections of steroid and a topical chloramphenicol and steroid cream mixture. Scarred or keloidal lesions may require surgery. PMID- 3048825 TI - AIDS in black patients. AB - Dermatologists function as consultants in the management of AIDS. They are consulted for care of cutaneous complications such as superficial infections (viral, dermatophyte, deep fungal, and bacterial). The management of these problems is similar to that in individuals with intact immune systems; however, these processes when associated with HIV infection may be more persistent and may recur. PMID- 3048826 TI - Sexually transmitted diseases in blacks. AB - The dermatologic findings indicative of STDs are invaluable clues to accurate diagnosis. The sexual consorts of confirmed and suspected STD patients must be promptly evaluated and treated of disease spread is to be curtailed. It is important to remember that some patients will present with more than one STD (Fig. 36). Sexually transmitted diseases in black patients may be atypical or exaggerated because of the host's unique tissue response. Every patient should be counseled regarding "safer sex." PMID- 3048827 TI - Pediatric dermatology in black patients. AB - An understanding of pediatric dermatology requires an insight into the differences between children and adults and a knowledge of the diseases that affect children exclusively. Therapy for the common dermatoses must be custom tailored for children to prevent unnecessary and potentially harmful exposure to medications. Black children have similar requirements but are further distinguished by the manner in which their skin reacts and expresses these eruptions. I hope that this presentation has provided a greater insight into the fascinating field of dermatology as it applies to black children. PMID- 3048828 TI - Cutaneous infections in blacks. AB - We have described briefly a variety of cutaneous infections seen commonly but not exclusively in blacks. Some disorders are prevalent in blacks because of geography (high temperatures, humidity), environment, and low socioeconomic status causing over-crowding, malnutrition, and poor or delayed access to medical care. Only a few occur as a result of the unique way black hair or skin responds to trauma or infection. Physicians managing black patients with dermatologic problems should be aware of and must consider all the factors that initiate, aggravate, and perpetuate cutaneous responses under these conditions. PMID- 3048829 TI - The old-age heart: normal aging changes which can produce or mimic cardiac disease. AB - The elderly segment of our society may triple by the year 2050. Specific cardiovascular data on the normal aging heart will be needed to provide proper medical and surgical therapy for this patient group. This report reviews normal aging changes of the very elderly heart. Expected or normal aging changes include brown atrophy of the myocardium, increased subepicardial fat, focal amyloid deposits, sigmoid-shaped ventricular septum, and calcific deposits in the aortic valve, mitral annulus and epicardial coronary arteries. Certain normal aging changes may produce clinical heart disease: aortic valve calcium (aortic stenosis), mitral valve annular calcium (mitral stenosis), amyloid deposits (amyloid heart disease). Certain normal aging changes may mimic heart disease: sigmoid-shaped ventricular septum (hypertrophic cardiomyopathy), mitral leaflet "buckling" (floppy mitral valve). PMID- 3048830 TI - The impact of the National Institutes of Health on the development of Canadian biomedical research. PMID- 3048831 TI - Hepatic photopenia in gallium imaging. AB - Although Ga-67 has been used extensively in imaging many different conditions, much is yet unknown about the mechanisms by which gallium concentrates preferentially in tissues such as the liver. This paper reports three patients with markedly diminished hepatic uptake of gallium. The cases are examined in light of published data demonstrating such factors as transferrin levels, radiation, and chemotherapeutic agents as altering the hepatic uptake of gallium. Of the many factors believed to influence the biodistribution of gallium, none can definitively account for the lack of hepatic uptake in these patients. PMID- 3048832 TI - Gallbladder visualization during technetium-99m RBC blood pool imaging. Case report and literature review. AB - Gallbladder visualization occurred after a Tc-99m red blood cell (RBC) cardiac gated blood pool scan. To date, seven cases of gallbladder visualization after the intravenous injection of Tc-99m RBCs have been reported. In the previous six patients the gallbladder was visualized incidentally during a search for gastrointestinal (GI) bleeding. All of the patients were anemic, six of seven had chronic renal failure, and five of seven had received multiple blood transfusions. When interpreting GI bleeding scans in patients with anemia and renal failure, awareness of the possibility of gallbladder visualization in the delayed images is important to avoid false-positive results. PMID- 3048833 TI - Pathophysiologic interpretation of time activity curves in dynamic bone imaging. AB - Reported here is a theoretical model based on a literature review correlating the 60 sec/25 min time activity curves (TAC) of dynamic bone imaging with the histologic components of bone. Information regarding healing versus nonhealing as well as a histophysiologic description of ongoing disease is obtainable from TACs of paired disease and nondiseased limbs. The TACs reflect the movement of Tc-99m MDP complexes across histologic compartments to reach the amorphous calcium phosphate (ACP) regions. These complexes must exit the bone capillaries, pass through the perivascular space, cross the osteoblastic barrier, enter the bone fluid space, and traverse collagen osteoid to reach ACP. The presence of diseases such as osteomyelitis, cellulitis, and degenerative arthritis and septic arthritis variously affect these spaces to cause typical perturbations in the TACs. The distinction of these patterns were observed in 48 patients, 12 per category. The model that explains how 60 sec/25 min TACs can reflect the histologic status of ongoing bone disease was applied to these disorders. PMID- 3048834 TI - Floating kidney. PMID- 3048835 TI - Caffey's disease: nuclear medicine and radiologic correlation: a case of mistaken identity. AB - Infantile cortical hyperostosis is declining in incidence. The classical radiograph and bone scan findings have been well described. A case first mistaken for child abuse is presented. The use of bone scanning in the work-up of non accidental trauma is addressed. PMID- 3048836 TI - Extrarenal abnormalities in Tc-99m DTPA renal perfusion studies due to hypervascularized tumors. AB - In cases of impaired renal perfusion and/or function, Tc-99m DTPA renal flow studies are often used as screening procedures. Besides the demonstration of pathological changes in renal perfusion and/or function, various extrarenal abnormalities may sometimes be detected. Three cases of hypervascular tumors were detected during the course of Tc-99m DTPA renography. These findings stimulated further diagnostic clarification revealing extended extrarenal lesions associated with several different etiologies: angiomyolipoma in tuberous sclerosis, extramedullary manifestations of plasma-cytoma, and metastasis of renal carcinoma. One should be aware of these extrarenal abnormalities in renal flow studies since these three tumor entities frequently exhibit increased vascularity. PMID- 3048837 TI - Detection of hepatic metastases in diffuse fatty infiltration by CT: the complementary role of imaging. AB - Six patients undergoing computed tomographic (CT) evaluation for possible abdominal and pelvic metastases were shown to have diffuse fatty infiltration of the liver and findings indeterminate for hepatic metastases. In two patients with diffuse fatty infiltration and no focal hepatic lesions on CT, technetium-99m sulfur colloid imaging demonstrated focal hepatic defects confirmed to represent metastases. In four patients with diffuse fatty infiltration and hyperdense liver foci on CT, radionuclide imaging demonstrated normal uptake in the hyperdense foci confirmed to represent areas of normal liver spared by fatty infiltration. In each of the six patients, clinical management was altered by the radionuclide findings. PMID- 3048838 TI - Accurate diagnosis of renal transplant rejection by indium-111 platelet imaging despite postoperative cyclosporin therapy. AB - Previous reports indicate that In-111 platelet scintigraphy (IPS) is a reliable test for the early diagnosis of acute post-operative renal transplant rejection (TR). However, the recent introduction of cyclosporin for post-transplantation immunosuppression requires that the diagnostic efficacy of IPS once again be established. Therefore, a prospective IPS study of 73 post-operative renal transplant recipients was conducted. Fourty-nine patients received cyclosporin and 24 patients did not receive this drug. Between these two patient groups, there were no significant differences in the diagnostic sensitivities (0.86 vs 0.80) and specificities (0.93 vs 0.84) with which TR was identified. We conclude that during the first two weeks following renal transplantation the cyclosporin treatment regimen used at our institution does not limit the reliability of IPS as a test for TR. PMID- 3048839 TI - Diagnosis of renal tubular transport disorders. A guide for the clinician. PMID- 3048841 TI - Familial achalasia, microcephaly, and mental retardation. Case report and review of literature. AB - Two Libyan brothers with achalasia of the cardia, microcephaly, and mental retardation are described. The parents were first cousins and were normal, as were two other male siblings and the female twin of one of the affected brothers. This is the second family to be reported with this constellation of abnormalities. The literature is reviewed for achalasia associated with other pathological conditions. PMID- 3048840 TI - Transient blindness following mild head trauma. Criteria for a benign outcome. AB - A series of seven children in Hawaii experienced transient cortical blindness following mild head trauma. All children, ages 3 through 8, recovered fully. The most prominent clinical feature was initial restlessness and agitation following relatively mild head trauma without significant loss of consciousness (LOC). One child may have experienced this several times. The clinical features associated with a benign outcome in this syndrome include: pediatric age group, mild head trauma, brief or no LOC, onset of blindness occurring within hours of the head injury, absent optokinetic nystagmus, duration of blindness less than 24 hours, agitation and restlessness, absence of skull fracture or visible cerebral injury on CT scan, absence of other neurological deficits, and EEG findings that initially show posterior slowing with subsequent normalization. Transiently fixed and dilated pupils have been described in these patients but should be viewed cautiously by clinicians in making this diagnosis, since cortical blindness is defined by sparing of the pupils. This syndrome may be underdiagnosed, since it may not be obvious that the child is blind unless the diagnosis is considered. PMID- 3048842 TI - Enhanced activation of the renin-angiotensin-aldosterone system in chronic cigarette smokers: a study of monozygotic twin pairs discordant for smoking. AB - Plasma renin activity (PRA) and aldosterone concentration were measured before and during submaximal exercise in 10 male monozygotic twin pairs who were discordant for smoking. In nine twin pairs PRA was higher in the smoker both at rest and during exercise. The mean PRA was 99% higher at rest and 84% higher during exercise than in nonsmokers. Plasma aldosterone levels were higher at rest in seven smokers and during exercise in eight smokers compared with the respective nonsmokers. The mean aldosterone level at rest was 23% and during exercise 40% higher in the smokers than in the nonsmokers. Chronic smoking induces increased PRA, which results in increased aldosterone formation, presumably via enhanced generation of angiotensin II. This may partly explain the greater vasoconstrictive reactivity typical of the arteries of chronic smokers. PMID- 3048843 TI - The 1988 AFCR Young Investigator Award for Clinical Research. PMID- 3048844 TI - Hormonal regulation of thyrotropin and gonadotropin gene expression. PMID- 3048845 TI - From receptors to genes--insights from molecular iron metabolism. PMID- 3048846 TI - In memory Kenneth Allan Platt, M.D. PMID- 3048847 TI - Health issues of Senior Citizens. PMID- 3048848 TI - Mitoxantrone: a novel anthracycline derivative. AB - The chemistry, pharmacology, pharmacokinetics, clinical efficacy, dosage and administration, and adverse effects of mitoxantrone are reviewed. Mitoxantrone, an aminoanthraquinone that was synthesized in 1979, belongs to a new chemical class of agents known as the anthracenediones. It possesses antiviral, antibacterial, immunomodulatory, and antitumor activity. The drug's antitumor activity is attributed to its interaction with DNA topoisomerase II, and its interaction with human cells may also involve nonintercalary, electrostatic interactions. Mitoxantrone is poorly absorbed orally and is most commonly administered intravenously. The drug is rapidly distributed into the red blood cells, white blood cells, and platelets, followed by deep-tissue sequestration. Mitoxantrone has demonstrated clinical efficacy in the treatment of leukemia, lymphoma, and breast cancer. As a single agent, mitoxantrone has a response rate of roughly 30% in acute nonlymphocytic leukemia or acute myeloid leukemia. In combination with other standard agents (cytarabine, vincristine, and prednisone), the response rate may reach 60%. In breast cancer, mitoxantrone's response rate as a single agent is 25-30%, while combination regimens produce response rates of 60% or more. The drug can cause cardiotoxicity with cumulative doses. Other adverse effects include myelosuppression, nausea and vomiting, stomatitis, mucositis, and alopecia. The cost of mitoxantrone is comparable to that of doxorubicin, but it is substantially more expensive than daunorubicin. Mitoxantrone is an important new agent with antitumor activity in leukemia, lymphoma, and breast cancer. In most situations, mitoxantrone will be considered second-line treatment or a restricted-use item because of its high cost and because of the lack of FDA approval for indications other than acute nonlymphocytic leukemia. PMID- 3048849 TI - Drug therapy of hypertensive crises. AB - The clinical features, pathogenesis, and pharmacologic management of hypertensive crises are reviewed, with emphasis on newer therapies. Hypertensive crises may be divided into hypertensive emergencies and hypertensive urgencies. Hypertensive emergencies, in which acute organ damage exists, require blood pressure reduction within one hour. In hypertensive urgencies no acute end-organ damage has yet occurred; however, blood pressure should be controlled within 24 hours. Factors that may precipitate a hypertensive crisis include renovascular hypertension, acute glomerulonephritis, head injuries, renin- or catecholamine-secreting tumors, antihypertensive-therapy withdrawal syndromes, eclampsia, and ingestion of tyramine by patients receiving monoamine oxidase inhibitors. The traditional drug of choice for therapy of hypertensive emergencies is sodium nitroprusside. Intravenous labetalol produces a prompt, controlled reduction in blood pressure and is a promising alternative. Other agents used are diazoxide, trimethaphan camsylate, hydralazine, nitroglycerin, and phentolamine. However, all these agents have disadvantages, including unpredictable antihypertensive effects, difficult blood pressure titration, and serious potential adverse effects such as profound hypotension, reduced renal blood flow, and increased myocardial workload. Most patients with hypertensive urgencies can be effectively treated with orally or sublingually administered agents. Older regimens of reserpine, methyldopa, or guanethidine, with their slow onsets and long durations of action, have been largely replaced by clonidine and nifedipine. Captopril and minoxidil have also been used with some success. Despite the lack of comparative trials with traditional agents, demonstrated efficacy and desirable pharmacologic characteristics have made several new agents acceptable for therapy of hypertensive crises. PMID- 3048850 TI - Reassessment of the value of lowering serum cholesterol. Questioning the wisdom of widespread intervention. PMID- 3048851 TI - Aminoglycoside pharmacokinetic monitoring: an integral part of patient care? PMID- 3048852 TI - Pisse prophecy: a brief history of urinalysis. AB - This article has reviewed a few of the major historic aspects of urine examination from ancient times to the twentieth century. It is a most fascinating history for it mirrored that of the history of medicine itself. The recognition of the importance of urine in diagnosis was made over 6000 years ago by several of the earliest civilizations, and a few of their clay tablets have been found that give us some insight into their observations and conclusions. However, the progress in analysis and its clinical significance was often slow and frequently littered with charlatanism. Visual inspection alone, and prognostication therefrom, was shown to be inadequate as a single means of diagnosis, and over 600 years ago Paracelsus, as well as others, began to reject ancient dogma and searched for new approaches to the analysis of urine by using chemical distillation techniques. Progress in anatomy and physiology, coupled with the understanding of organ function, provided a fertile field for the investigation of the composition of the urine and associating its chemical constituents with specific disease states. The advent of the microscope caused scientists to examine all body fluids, especially the urine, and to record their observations as an aid to diagnosis--the beginnings of medical microscopy. This century has also seen remarkable advances in the field of urinalysis: dipstick testing, the application of modern chemical and microscopic techniques to constituent analysis, automation, and, most recently, monoclonal antibody and recombinant gene technology to enhance and improve urine examination. In short, urinalysis, the first of all laboratory tests, began as and still remains a most valuable and highly important means of diagnosis in clinical medicine. PMID- 3048853 TI - Quality assurance in urinalysis. AB - Instituting a Quality Assurance program in urinalysis is an important step in achieving accurate and reproducible results. This article has explored the various components of such a program for routine urine examination and included a discussion, along with "how to" recommendations, of such topics as specimen collection and handling, dipstick chemical analysis, sediment evaluation, and reporting. Properly collected fresh urine samples should be analyzed promptly, preferably without refrigeration. Each step of the urinalysis procedure should be carefully controlled and standards applied to assure optimal results. Reports should include clinically useful pertinent data organized in convenient, readable fashion. PMID- 3048854 TI - Monoclonal antibodies: use in urine sediment examination. AB - Accurate identification of nucleated cells in urine can be difficult with conventional methods of microscopic urinalysis. Monoclonal antibodies were used with an immunoperoxidase technique to identify nucleated cells in urine. This new development in urinalysis is in its early stages, but it has helped to circumvent the difficulties associated with standard microscopy. The monoclonal antibody technique allowed for the identification of granulocytes, monocytes, lymphocytes, glomerular epithelial, proximal tubular, loop of Henle, distal tubule/collecting duct, and urothelial cells in urine, and by quantifying these cells it was possible to determine the urine cell profiles in various renal diseases as well as in allograft rejection and early post-transplant acute tubular necrosis in renal allograft recipients. The cell profiles are useful in aiding the diagnosis of these conditions. PMID- 3048855 TI - Urinary parameters to assess renal function. AB - Acute suppression of renal function has a multitude of etiologies. This review discusses the pathophysiologic basis, usefulness, and limitations of the commonly utilized urinary function parameters as well as some miscellaneous tests to properly diagnose specific disorders. PMID- 3048856 TI - Proteinuria. AB - Proteinuria is one of the most common manifestations of renal disease. Changes in plasma protein concentration, glomerular permeability, tubular reabsorption, or renal hemodynamics can affect the rate of urinary excretion and the composition of urinary proteins. Determination of the significance of the proteinuria requires an evaluation of the amount and type of proteinuria in the context of the clinical setting. In addition, the clinician must be familiar with the laboratory evaluation of proteinuria and its limitations in order to correctly interpret these tests. PMID- 3048858 TI - Urinalysis in the diagnosis of urinary tract infections. AB - Urinary tract infection is the commonest human bacterial infection. Bacteriuria alone does not appear to produce progressive renal damage or hypertension. However, it can produce considerable morbidity. Urinalysis is a simple, relatively sensitive, and reliable way of diagnosing urinary tract infection. It is not clear that routine screening should be performed in all patients, but pregnant females, patients with known anatomic abnormalities, and patients with recent genitourinary instrumentation should be screened. The major determinant of therapeutic success in patients with urinary tract infections is the anatomic site of infection. Superficial mucosal infection of the bladder is well treated with a single dose of an appropriate antibiotic, whereas deep tissue infection of the kidney or prostate should be treated with a prolonged and intensive course of therapy. Urinalysis is an insensitive tool in the localization of infection. However, the presence of white cell casts on the examination of the urinary sediment is pathognomonic of upper tract infection and would lead one to pursue an aggressive course of therapy. Examination of the concentrating ability is of limited help in this regard because of the wide range of overlap of concentrating ability in patients with upper and lower tract infections. In selected instances, urinalysis is of help in guiding therapy of urinary tract infections. This is particularly true of the patients with acute urethral syndrome where therapy is guided by the presence or absence of pyuria. Urinalysis, a simple front-line test, is of paramount importance in the evaluation and management of the patient with urinary tract infection. PMID- 3048857 TI - Flow cytometry of urine. AB - Flow cytometry is becoming an increasingly important tool for the evaluation of urine cytology. Although it has established a strong foothold in oncologic DNA analysis, studies of whole cells and cell markers are only in their infancy. In addition, potential diagnostic applications such as those involving binding of E. coli strains to uroepeithelial cells will move flow cytometry from a research tool to a real diagnostic tool. PMID- 3048859 TI - An algorithm for hematuria. AB - Two algorithms are presented to help clarify the diagnostic evaluation of hematuria. Utilizing an anatomic framework for adults and an etiologic framework for children, this complex clinical problem can be approached in a logical manner. PMID- 3048860 TI - Microalbuminuria. AB - Nephropathy progressing to end-stage renal failure is a common complication of diabetes mellitus. The development of diabetic nephropathy is preceded for many years by minor increases in urine albumin excretion (microalbuminuria), and some diabetologists believe that by reducing or reversing this early abnormality, possibly by more rigorous control of hyperglycemia or by correcting glomerular hemodynamic abnormalities, the later development of frank nephropathy might be delayed or prevented. Once frank nephropathy has developed, its progress to renal failure is little, if at all, influenced by therapy. The early detection of microalbuminuria is therefore important for the management of diabetic patients, and its measurement should be available to all diabetologists. Sensitive, precise, and accurate methods are readily available to most laboratories. The preferred specimen is a 24-hour collection of urine or a timed overnight collection. A quantitation corrected for creatinine on a first-morning specimen may be a sufficiently sensitive test to detect those patients who should be subjected to a more extensive collection. Whether random samples taken at other times of the day are also suitable for screening remains to be proved. PMID- 3048861 TI - Demonstration of cross-reactions between pneumococci and alpha-streptococci using gold-labelled mono- and polyclonal antibodies and electron microscopy. AB - Cross-reactions between alpha-streptococci and the pneumococcal C-polysaccharide (PnC) were investigated using electron microscopy and immunogold labelling of bacterial cells. Monoclonal antibodies against two different determinants of the PnC molecule were used, one directed against the chain sugar of the repeating unit 2-acetamido-4-amino-2,4,6-trideoxygalactose (Sug) and the other against the phosphorylcholine residue. Two different immunogold techniques were tested, either by direct labelling of the monoclonal antibody or by using an antimouse immunogold conjugate to demonstrate binding of the monoclonal antibodies. Antibodies against the "Sug" determinant reacted only with pneumococci, whereas antibodies against the phosphorylcholine determinant bound to cross-reacting streptococci as well as to pneumococci. These results indicate that the cross reacting antigens of the alpha-streptococci contain phosphorylcholine residues, but that they are not identical to the C-polysaccharide molecule. PMID- 3048862 TI - Less severe hypertension--when and how it should be treated. PMID- 3048864 TI - Etiology, diagnosis, and management of congenital cardiac disorders. PMID- 3048863 TI - Management of ventricular arrhythmias. AB - Left ventricular dysfunction and complex VPCs are independent risk factors for subsequent cardiac death in patients with ischemic or nonischemic heart disease. The use of antiarrhythmic drugs during acute myocardial infarction is discussed. Patients with complex VPCs associated with heart disease should be treated with antiarrhythmic drugs. Drug efficacy should be evaluated by a 24-hour ambulatory ECG recording and by a treadmill exercise stress test. Blood drug levels should be measured at appropriate times. Maintenance doses of the various antiarrhythmic drugs are listed. Electrophysiologic testing with induction of ventricular tachycardia by extrastimulation may predict the clinical efficacy of the antiarrhythmic drug or combination of drugs needed for the long-term treatment of ventricular tachyarrhythmias. Electrophysiologic testing must be used for selecting antiarrhythmic drugs in patients who have experienced sustained ventricular tachycardia or ventricular fibrillation but who are free of VPCs during ambulatory ECG monitoring and exercise stress testing. If patients have life-threatening ventricular tachycardia or fibrillation resistant to antiarrhythmic drugs, surgical intervention is indicated. Complex VPCs in asymptomatic patients without heart disease should not be treated with antiarrhythmic drugs. PMID- 3048865 TI - Controlling cholesterol levels through exercise. PMID- 3048866 TI - Chronic orthostatic hypotension. PMID- 3048868 TI - A performance evaluation of the expert system ANEMIA. AB - This paper reports the results of an evaluation study of the current level of performance given by ANEMIA, a knowledge-based consultation system addressing the clinical problem of managing anemic patients. ANEMIA was developed on a mainframe using the AI programming scheme EXPERT and then translated into a version running on a personal computer. At present the system is able to provide assistance in the diagnosis and management of 65 disease entities. After extensive local testing of accuracy, completeness, and consistency of the knowledge base included into ANEMIA, we designed a study to evaluate whether the system is able to appropriately mirror also the reasoning of well-known hematologists other than those who provided the knowledge. We were also interested in testing whether there were conflicting opinions among hematologists. Thus, we designed a validation study in which ANEMIA's performance could be compared with that of six hematologists and the interexpert consensus evaluated. ANEMIA's overall performance was judged acceptable in 87% (26/30) of the cases, while expert evaluators agreed with their colleagues in 90% (27/30) of them. A low interexpert consensus was found: considering the ratings given by different hematologists to the same ANEMIA performance, complete agreement occurred only 47% of the time. PMID- 3048869 TI - Consumer development summeries. Cochlear implants. National Institutes of Health. PMID- 3048867 TI - Coronary heart disease in the elderly. AB - In 1984, 435,759 deaths were attributed to CHD among persons greater than or equal to 65 years of age. CHD was the leading cause of death in this group. Death rates rose steeply with age among the elderly. Men had higher death rates than women, but the male-to-female ratio declined with increasing age. Considerable geographic variation in CHD mortality in the elderly was noted. Since 1968, CHD death rates have declined in persons greater than or equal to 65 years of age in each age, sex, and race group. However, prevalence of self-reported CHD in the elderly population has increased. Prevalence rates increased with age except for a slight decrease above age 75 in men. In 1985, 436,000 persons aged greater than or equal to 65 years were discharged with a principal diagnosis of acute MI. The hospital case fatality rate was 21.8%. Since 1970, hospitalization rates for acute MI have generally increased, while hospital fatality rates have decreased for persons greater than or equal to 65 years of age. Since 1979, utilization of coronary artery bypass surgery and coronary arteriography have dramatically increased among the elderly. In 1980 and 1981, elderly persons made six million visits to physicians' offices for chronic CHD. CHD contributed importantly to the 1980 expenditures of 3.3 billion dollars in men and 4.8 billion dollars in women greater than or equal to 65 years of age for heart disease care. Although mortality rates from CHD in the elderly have decreased since 1968, increasing hospitalization rates and utilization of other health care services emphasize the need for more vigorous efforts at prevention.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3048870 TI - Consequences of uterine blood loss caused by various intrauterine contraceptive devices in South American women. World Health Organization Special Programme of Research, Development and Research Training in Human Reproduction. AB - Increased menstrual blood loss (MBL) associated with intrauterine device (IUD) use may precipitate or aggravate iron deficiency anaemia, adversely affecting the health of women particularly those from developing countries. Studies were conducted to define the association of MBL and iron status in South American women; to determine the level of MBL induced by IUD use which would result in iron depletion, the length of time for this depletion to occur and, comparing various IUDS, to determine if any currently tested IUDs are suited to long-term use in South American women. A total of 395 women received one of 5 types of IUDs in Santiago, Chile, and Juiz de Fora, Brazil: Lippes Loop, Multiload-250 and Multiload-375 were used in both centres; in Santiago some subjects received the Copper-7 or ProgestasertR devices and in Juiz de Fora, the TCu 200 and the T Chloroquin IUDs were also tested. MBL and haemoglobin (HGB) were measured for 3 menstrual cycles before insertion, and following insertion, at one, two, four, six, nine, twelve, eighteen and twenty-four months in the majority of cases. Serum ferritin was measured before insertion and at intervals of six months. Mean values of MBL prior to IUD insertion in both centres varied from 21-30 ml. As with previous publications, the use of the Lippes Loop was associated with the greatest increase in MBL which was sustained throughout the 24 months of observation. Women who had one of the two types of Multiload devices inserted also had increased MBL and reduced ferritin for at least 12 months of use. TCu 200 and Copper-7 IUD users had an initial increase in MBL of 1 to 17 ml in the first six months of observation returning to normal levels beyond six months. Serum ferritin levels were lower for one year and then returned to admission values. ProgestasertR users confirmed previous reports of a reduction of 40-50% in MBL and an increase in serum ferritin. Few significant changes in haemoglobin (HGB) concentrations were found. Serum ferritin levels on admission ranged from 7.1 to 16.4 ng/ml in Santiago and from 15.8 to 23.2 ng/ml in Juiz de Fora. Many women were in a marginal state of iron balance as evidenced by lower serum ferritin values. Changes in serum ferritin were very closely related to those in MBL.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3048872 TI - A bibliography of the statistical analysis of vaginal bleeding patterns. AB - An annotated bibliography of 123 references and review papers on graphical description and statistical analysis of vaginal bleeding patterns is presented. PMID- 3048871 TI - The analysis of menstrual bleeding patterns: a review. AB - Patterns of vaginal bleeding are an important factor in the acceptability of contraceptive methods. The analysis of data obtained from daily menstrual diary records is a major methodological problem to which no satisfactory solution exists. This review describes approaches to the analysis of bleeding patterns among contracepting and non-contracepting women and reviews the difficulties involved. The reference period method, introduced to avoid the arbitrary rules and definitions required for an analysis based on the concept of a menstrual cycle, is discussed and its limitations presented. The review draws on reports of meetings convened by the World Health Organization and the University of Exeter Family Planning Unit to discuss issues in the analysis. Previously unpublished methods are summarized and areas of controversy and topics for further research are identified. PMID- 3048873 TI - Carnitine metabolism in chronic renal failure. Acute effects of carnitine addition to the dialysate during acetate hemodialysis. PMID- 3048874 TI - Lipid metabolism in patients with chronic renal failure in the predialytic phase. AB - CRF is accompanied by characteristic alterations of lipoprotein metabolism with a retarded catabolism of triglyceride-rich lipoproteins as a prominent feature. Further investigation of the lipoprotein profile of CRF could provide information on the fundamental pathophysiological processes responsible for these disturbances and their possible clinical significance. PMID- 3048876 TI - Comparative analysis of structural change in a free autovenous graft and in a transplant with spiral reinforcement after correction of experimental arterial defects. AB - In sixty-four dogs, a comparative analysis was made of structural changes produced in the wall of free autovenous graft and of a transplant strengthened by a spiral reinforcement (lavsan monothread) after transplantation of femoral vein segments into the common carotid artery. Histological and morphometric examinations made in an interval ranging from 3 days to 1 1/2 years after surgery showed that the strengthening of the venous graft by means of a reinforcement prevents its overextension when functioning as an artery, and keeps the endothelium intact. This reduces the danger of thrombotic complications in the operation area. The spiral reinforcement at the same time prevents compression of the vein by surrounding cicatricial tissue as well as an aneurysmatic extension of the transplant. PMID- 3048877 TI - Non-pharmacological treatment of hypertension. A review. PMID- 3048878 TI - Status of echocardiography in clinical cardiology. AB - Echocardiography (ECHO) is at present the most important and most useful non invasive diagnostic method employed in cardiology. Its dominant position is the result of dramatic development of its technology and methodology enabling a wide range of clinical applications. Apart from M-mode and two-dimensional ECHO, these methods include mainly Doppler ECHO (pulse and continuous, colour), computer, contrast, exertional, transoesophageal ECHO, ultrasound characteristics of tissue (textural analysis). Moreover, technical improvements have enabled to quantify, standardise and unify ECHO measurements and nomenclature. The most general contribution of ECHO to present-day clinical cardiology lies in the confirmation or exclusion of a suspected cardiovascular disease and assessment of the degree of circulatory system involvement. PMID- 3048879 TI - Doppler echocardiography in the diagnosis of pulmonary hypertension. AB - Review of the current state of knowledge about Doppler diagnosis of pulmonary hypertension is presented. The authors compare the findings of numerous studies with their own data and analyse the causes of discrepancies and controversies that have remained to date. Doppler estimation of mean pulmonary artery pressure (PAP) using acceleration time (ACT) measurement in the right ventricular outflow tract or main pulmonary artery has been shown to correlate closely with simultaneously measured values of PAP both at rest and during exercise (r = -0.92 and -0.94, respectively). ACT values show sufficient sensitivity for the detection of pulmonary hypertension (89%), at maximum (100%) specificity. However, Doppler values not obtained simultaneously do not necessarily correlate with PAP in view of rapid physiological variations of PAP. For everyday diagnostic practice in cardiology, prediction based upon mere qualitative estimation of the type of velocity curve seems sufficient. Doppler diagnosis of pulmonary hypertension, little demanding on both technical skills and equipment, is gaining a priority status among the non-invasive methods available. PMID- 3048875 TI - Vitamin status in patients with chronic renal failure. AB - Many factors complicate the effort for a recommendation on individual vitamin requirements in CRF. On the basis of our present incomplete knowledge about the handling of vitamins in uremia, suggestions for appropriate supplementation only of water-soluble vitamins are given. Patients with advanced CRF without dialysis treatment should receive daily supplements of vitamin B6 (5 mg), ascorbic acid (70-100 mg), and the normal recommended daily allowance of the other water soluble vitamins in addition to the vitamin intake from the diet. We give folic acid only in patients taking antifolate drugs or in combination with iron in iron deficiency state and anemia (1 tablet of Folicombin contains 0.5 mg folic acid and 0.4 g elemental iron). There is still a pressing need for more data on the vitamin status, on vitamin requirements, and on long-term effects of vitamin administration in CRF. PMID- 3048880 TI - Keratoconus. AB - Keratoconus is a bilateral disorder of corneal shape which may be sporadic or genetically determined. Early corneal thinning suggests that a functional loss of structural elements is a primary event in the disease. Tensile strength of the cornea is reduced and is expressed by signs of rupture and scarring in Bowman's layer, scarring in the substantia propria, and rupture of Descemet's membrane. The overall stretching of the cornea results in an increase in curvature while an increased area of the corneal surfaces probably determines the onset and form of Fleischer's ring and the occurrence of endothelial polymegathism. Biochemical studies have shown an increase in collagenolysis and of reduceable collagen cross links, but there is inconsistent evidence of altered solubility or of hydroxyproline or proteoglycan content. Of recent interest is the characterization of proteoglycan bridges along and between corneal collagen fibrils in keratoconus and the apparent loss of keratan sulphate demonstrated by electron-histochemical and x-ray diffraction techniques. The manner in which this could interfere with corneal strength is discussed. PMID- 3048881 TI - Imaging of the cornea. AB - The aim of the LSU Eye Center Corneal Topography Project was to apply computer analysis to keratoscopy and develop graphic presentation methods to provide accurate and easily assimilated information regarding the status of patient corneal topography. The analysis contains four diagrams: a plot of the raw keratoscope data for quality control, a three-dimensional wire model of corneal surface power distribution, a plot of dioptric point surface powers, and a color coded contour map of corneal surface powers. While the analysis uses a current generation 11 ring keratoscope for producing keratoscope photographs, it can be adapted to newer keratoscopes, which promise to give broader coverage of the corneal surface. Examples of normals and several surgical patients will be shown, including cases in which the method has pointed to potential refinements in the theory and application of refractive surgical techniques. PMID- 3048882 TI - The clinical measurement of endothelial permeability. AB - The endothelium maintains corneal deturgescence in part by serving as a barrier to fluid movement into the cornea. This barrier function is estimated by measuring the permeability of the endothelium to small, nontoxic, fluorescent molecules such as fluorescein and carboxy-fluorescein. After topical or systemic application of the dye, its concentration in the cornea and the anterior chamber is measured noninvasively at intervals over several hours with a fluorophotometer. Endothelial permeability to the fluorescent dye is calculated from the changes in its concentration in the cornea and anterior chamber over time. The permeability of the endothelium to the small fluorescent molecules is assumed to be proportional to its permeability to water, and thus provides an estimate of the barrier function of the endothelium. This paper reviews the theory and technique of clinical fluorophotometry and discusses published data concerning the effects on endothelial permeability of aging, contact lens wear, medication, and surgery. PMID- 3048883 TI - Atypical corneal dystrophy with stromal amyloid deposits. AB - Corneal dystrophies are distinctive clinically and histopathologically; however, variations do occur. We present two cases of stromal amyloidosis from one family masquerading other corneal dystrophy. The two cases are from a six-generation family with an autosomal dominant corneal dystrophy resembling Reis-Bucklers' dystrophy. In these cases, neither the propositus nor other family members showed typical lattice lines. Light and electron microscopy of the obtained corneal buttons disclosed amyloid stromal deposits. The clinicopathologic correlation of these cases suggests that this family represents a variant of stromal amyloid dystrophies. PMID- 3048884 TI - Visual loss following suture removal postkeratoplasty. AB - Suture removal following penetrating keratoplasty continues to plague surgeons as one of the most unpredictable aspects of this procedure. The patients presented here will detail three cases of endophthalmitis and one case of expulsive hemorrhage following suture removal. These events cannot be predicted. However, certain steps can be taken to minimize these potentially blinding complications. These steps include avoidance of through and through sutures, follow-up examination within 24 h of suture removal to look for signs of would leak, graft dehiscence or infection, use of appropriate antibiotics, elimination of dacryocystitis prior to keratoplasty, and particular vigilance in immunocompromised patients. PMID- 3048885 TI - Clinical pathology of non-freeze lamellar refractive keratoplasty. AB - We studied two corneal specimens obtained following corneal transplantation for loss of best corrected vision after planar lamellar refractive keratoplasty. The epithelium appeared slightly undifferentiated in both cases. Peripheral bends and occasional breaks were found in the periphery of Bowman's layer. In one case there were areas of subepithelial fibrosis as well as ultrastructural fractures in Bowman's layer. The keratocyte population appeared to be slightly decreased in one case. In both specimens the optical interface contained active fibroblasts in the periphery. The planar lamellar refractive keratoplasty technique theoretically eliminates many of the adverse morphologic features encountered following the standard Barraquer cryolathe techniques, but clinical studies will be needed to determine if this form of lamellar keratoplasty is clinically superior to currently practiced techniques. PMID- 3048886 TI - A new surgical technique for posterior chamber lens fixation during penetrating keratoplasty in the absence of capsular or zonular support. AB - A new technique has been developed to implant or exchange an anterior chamber or iris supported lens with a posterior chamber lens during penetrating keratoplasty. The posterior chamber intraocular lens is suspended by suturing the haptics into the ciliary sulcus rather than to the iris. PMID- 3048887 TI - Enzymes of nucleotide metabolism: the significance of subunit size and polymer size for biological function and regulatory properties. AB - The 72 enzymes in nucleotide metabolism, from all sources, have a distribution of subunit sizes similar to those from other surveys: an average subunit Mr of 47,900, and a median size of 33,300. The same enzyme, from whatever source, usually has the same subunit size (there are exceptions); enzymes having a similar activity (e.g., kinases, deaminases) usually have a similar subunit size. Most simple enzymes in all EC classes (except class 6, ligases/synthetases) have subunit sizes of less than 30,000. Since structural domains defined in proteins tend to be in the Mr range of 5,000 to 30,000, it may be that most simple enzymes are formed as single domains. Multifunctional proteins and ligases have subunits generally much larger than Mr 40,000. Analyses of several well-characterized ligases suggest that they also have two or more distinct catalytic sites, and that ligases therefore are also multifunctional proteins, containing two or more domains. Cooperative kinetics and evidence for allosteric regulation are much more frequently associated with larger enzymes: such complex functions are associated with only 19% of enzymes having a subunit Mr less than or equal to 29,000, and with 86% of all enzymes having a subunit Mr greater than 50,000. In general, larger enzymes have more functions. Only 20% of these enzymes appear to be monomers; the rest are homopolymers and rarely are they heteropolymers. Evidence for the reversible dissociation of homopolymers has been found for 15% of the enzymes. Such changes in quaternary structure are usually mediated by appropriate physiological effectors, and this may serve as a mechanism for their regulation between active and less active forms. There is considerable structural organization of the various pathways: 19 enzymes are found in various multifunctional proteins, and 13 enzymes are found in different types of multienzyme complexes. PMID- 3048888 TI - Spectrin and related molecules. AB - This review begins with a complete discussion of the erythrocyte spectrin membrane skeleton. Particular attention is given to our current knowledge of the structure of the RBC spectrin molecule, its synthesis, assembly, and turnover, and its interactions with spectrin-binding proteins (ankyrin, protein 4.1, and actin). We then give a historical account of the discovery of nonerythroid spectrin. Since the chicken intestinal form of spectrin (TW260/240) and the brain form of spectrin (fodrin) are the best characterized of the nonerythroid spectrins, we compare these molecules to RBC spectrin. Studies establishing the existence of two brain spectrin isoforms are discussed, including a description of the location of these spectrin isoforms at the light- and electron-microscope level of resolution; a comparison of their structure and interactions with spectrin-binding proteins (ankyrin, actin, synapsin I, amelin, and calmodulin); a description of their expression during brain development; and hypotheses concerning their potential roles in axonal transport and synaptic transmission. PMID- 3048889 TI - Transcription elements and factors of RNA polymerase B promoters of higher eukaryotes. AB - The promoter for eukaryotic genes transcribed by RNA polymerase B can be divided into the TATA box (located at -30) and startsite (+1), the upstream element (situated between -40 and about -110), and the enhancer (no fixed position relative to the startsite). Trans-acting factors, which bind to these elements, have been identified and at least partially purified. The role of the TATA box is to bind factors which focus the transcription machinery to initiate at the startsite. The upstream element and the enhancer somehow modulate this interaction, possibly through direct protein-protein interactions. Another class of transcription factors, typified by viral proteins such as the adenovirus EIA products, do not appear to require binding to a particular DNA sequence to regulate transcription. The latest findings in these various subjects are discussed. PMID- 3048891 TI - Some futuristic possibilities for resuscitation. AB - Future possibilities for resuscitation must take into account our still limited understanding of the reperfusion syndrome. Clinical resuscitation research must incorporate the use of prolonged life-support techniques. These may depend on the use of cardiopulmonary bypass to provide improved reperfusion of vital organs and to permit the time necessary to evaluate and treat the mediators and modifiers of the reperfusion syndrome. It is likely that some patients who require prolonged life support will need replacement organs or, should their brains fail to survive resuscitation, become organ donors. Physicians involved in resuscitation and transplantation must come to grips with the logistic problems of techniques for prolonged resuscitation. PMID- 3048890 TI - Scientific evaluation of clinical interventions in resuscitation medicine. AB - The image of the physician as a therapist, as well as the general perception of the physician's role in society and in the life of patients and their families, has changed dramatically in past decades. In response, many conferences, editorials, and planning activities for medical and premedical curricula have been devoted to the subject of restoring the elements of compassion and personal involvement to the relationship between physician and patient. Nonetheless, the change in the doctor-patient relationship might merely reflect the growing indifference to people as individuals that seems to pervade our society in all service-related areas. Although the loss is particularly objectionable for the sick, some may claim it is compensated by scientifically superior care in present day medical practice. This paper will reflect briefly on past and present views of medical care and its evaluation. It will conclude that a scientifically sound evaluation of treatment might also offer a solution for restoration of human elements in the delivery of medical care. PMID- 3048892 TI - Ethical dilemmas in organ donation and transplantation. AB - Since the first successful organ transplantation in 1953, we have seen an explosive development in transplantation surgery, particularly during the 1980s. With it followed an abundance of legal controversies and ethical dilemmas. Optimal use of viable organs necessitated precise definition of brain death in heart-beating cadavers with artificially maintained ventilation and circulation. Viable organs must remain well perfused to be suitable for procurement and transplantation into carefully selected recipients on an equal-opportunity basis. Due consideration must be given to both medical and social indications. At present, homografts dominate the field of organ transplantation; however, because of the shortage of human organs, both artificial organs (especially hearts) and xenografts are expected to become increasingly common in the near future. No doubt, the use of such modern technology will introduce additional ethical problems. PMID- 3048893 TI - Philosophical, ethical, and legal aspects of resuscitation medicine. I. Deferred consent and justification of resuscitation research. AB - Informed prospective consent for clinical resuscitation research may not be possible. Deferred consent is an untenable notion. Consent to continue in research cannot be used to support a claim that there was, or would have been, consent to the initiation of research. The conditions for the justifiability of resuscitation research without informed consent are: a) patient is comatose; b) lifesaving treatment must be given immediately; c) given all available evidence, there is reason to believe that the probability of death or severe deficit with experimental or control therapy is not greater than the probability of death or severe deficit on usual therapy; d) given all available evidence, there is reason to believe that the probability of normal or near-normal outcome is greater on experimental or control therapy than on usual therapy; and e) the study can provide evidence on whether there is a significant difference between experimental and control therapies in the incidence of normal or near-normal survival. PMID- 3048895 TI - Postresuscitation disease. AB - The postresuscitation disease is a specific pathophysiologic state of vital organ systems early after ischemic anoxia. This report summarizes reviews of past research and makes suggestions for future research concerning revival of the cerebral cortex after clinical death, CNS stimulation vs. sedation, postischemic coma and pain, near-death experiences, and extracerebral derangements. The stages of resuscitation when the CNS should be stimulated and those when it is preferable to depress the activity of not fully recovered higher centers remain to be clarified. Future research in reanimatology should include the chemical nature of endotoxins in terminal states. Adult respiratory distress syndrome (ARDS, shock lung), a component of the postresuscitation disease, occurs frequently after cardiac arrest or in sepsis and cannot be fully prevented by artificial ventilation. Prevention of ARDS should also be studied. PMID- 3048894 TI - Resuscitation from clinical death: pathophysiologic limits and therapeutic potentials. AB - Modern cardiopulmonary-cerebral resuscitation (CPCR) for the reversal of clinical death (i.e., prolonged cardiac arrest) is a sequence of basic, advanced and prolonged life-support steps. This system was initiated by research that started in the 1950s. Present community-wide results are encouraging, but suboptimal. Maximal benefit from CPCR will be achievable: a) by minimizing response times; and b) by extending reversible arrest times--the topic of this symposium. For reperfusion, closed chest CPR is more readily available than, but physiologically inferior to, open chest CPR and emergency cardiopulmonary bypass. To optimize outcome, four components of the postresuscitation syndrome are being investigated: a) perfusion failure; b) reoxygenation injury cascades; c) self intoxication; and d) blood derangements. Results from animal outcome studies so far suggest significant but still inconsistent benefit from several special postarrest treatments. The longest normothermic no-flow time yet reversed to good functional survival of heart, brain and the entire organism appears to be not 5 min, but between 10 and 20 min. The following is recommended and in part has been initiated: a) simultaneous investigation of pathophysiologic limits, therapeutic potentials, and prognosticating measurements; b) simultaneous basic research at cellular, organ, and organism levels; c) increased communication and consensus on research models between research centers; d) use of short-term and long-term animal models for systematic mechanism-oriented and empirical outcome-oriented studies; e) development of etiology-specific combination treatments; and f) community-wide case registries combined with epidemiologic studies and randomized clinical treatment trials. PMID- 3048896 TI - Mechanisms of ischemic brain damage. AB - This article provides a brief review of recent developments regarding the pathophysiology of ischemic brain damage, and offers hypotheses explaining the pathogenesis of selective neuronal vulnerability and of tissue infarction, respectively. It is suggested that selective neuronal vulnerability, observed after brief periods of ischemia and after hypoglycemic coma, qualifies as an excitotoxic lesion, which causes postsynaptic damage to neurons innervated by excitatory amino acids by enhancing calcium influx. However, ischemic damage often involves glial and vascular cells as well, and causes infarction. It is hypothesized that this type of brain damage is related to acidosis and that enhanced acidosis is detrimental because it accelerates delocalization of protein bound iron, with an ensuing free-radical damage to membrane lipids and proteins. PMID- 3048897 TI - Morphology of global cerebral ischemia. AB - The cerebral tissue responses to ischemia are modified by many factors, including the mechanism by which ischemia is induced, and the severity and duration. Children can withstand brain ischemic insults that are usually lethal to older patients. Other modifying factors of ischemia, such as barbiturates and serum glucose, remain poorly defined in humans. Once a cerebral ischemic event occurs, for example, after cardiac arrest and resuscitation, secondary events (such as alterations in the perfusability and reactivity of the microcirculation) may aggravate the original ischemic injury at discrete sites of the brain. A more exact definition of a) the limits of reversibility of ischemic injury, b) the histologic features of sublethally injured tissues, and c) the effects of various therapeutic interventions, awaits the development of relatively inexpensive and reproducible animal models of ischemia in which objective comparisons can be made between the nature of the local circulatory conditions and the accompanying histologic abnormalities. PMID- 3048898 TI - Complete global myocardial ischemia in dogs. AB - Complete global myocardial ischemia or zero coronary arterial flow in dogs results in a series of well-defined changes which begin when the myocardium converts from aerobic to anaerobic metabolism. These processes continue until the myocardium dies. The products of anaerobic metabolism, chiefly glycolytic intermediates, inorganic phosphate, H+, and creatine, are produced intracellularly and accumulate in the tissue. Because the demand for high-energy phosphates (HEP) in the tissue exceeds the supply available from anaerobic glycolysis and HEP reserves, net ATP level declines, approaching zero after 100 min of ischemia at 37 degrees C. At this time, the changes in totally ischemic tissue in vitro are equivalent to those seen in myocytes irreversibly injured by severe ischemia in vivo. During reoxygenation after total ischemia, the resumption of effective contractile activity depends partly on the metabolic changes and degree of myocyte injury sustained during ischemia. PMID- 3048899 TI - Optimization of oxygen transport in mechanically ventilated newborns using oximetry and pulsed Doppler-derived cardiac output. AB - In respiratory distress syndrome (RDS), PEEP improves arterial oxygenation but may impair cardiac output. The effects of PEEP on gas exchange and hemodynamics were studied in 12 mechanically ventilated newborns in the acute phase of RDS. Stepwise increase in PEEP resulted in both a) a progressive increase in PaO2 and transcutaneous oxyhemoglobin saturation, and b) a depression of pulsed Doppler measured cardiac output that was statistically significant at 9 cm H2O PEEP. Thus, averaged systemic oxygen delivery (DO2) was maintained with improved arterial oxygenation up to 6 cm H2O PEEP. Further increase in PEEP induced a significant fall in DO2. No variation was observed in heart rate and mean arterial pressure. The combined use of oximetry and pulsed Doppler echocardiography enables noninvasive optimization of mechanical ventilation and PEEP during the clinical course. PMID- 3048900 TI - Pediatric risk of mortality (PRISM) score. AB - The Pediatric Risk of Mortality (PRISM) score was developed from the Physiologic Stability Index (PSI) to reduce the number of physiologic variables required for pediatric ICU (PICU) mortality risk assessment and to obtain an objective weighting of the remaining variables. Univariate and multivariate statistical techniques were applied to admission day PSI data (1,415 patients, 116 deaths) from four PICUs. The resulting PRISM score consists of 14 routinely measured, physiologic variables, and 23 variable ranges. The performance of a logistic function estimating PICU mortality risk from the PRISM score, age, and operative status was tested in a different sample from six PICUs (1,227 patients, 105 deaths), each PICU separately, and in diagnostic groups using chi-square goodness of-fit tests and receiver operating characteristic (ROC) analysis. In all groups, the number and distribution of survivors and nonsurvivors in adjacent mortality risk intervals were accurately predicted: total validation group (chi 2(5) = 0.80; p greater than .95), each PICU separately (chi 2(5) range 0.83 to 7.38; all p greater than .10), operative patients (chi 2(5) = 2.03; p greater than .75), nonoperative patients (chi 2(5) = 2.80, p greater than .50), cardiovascular disease patients (chi 2(5) = 4.72; p greater than .25), respiratory disease patients (chi 2(5) = 5.82; p greater than .25), and neurologic disease patients (chi 2(5) = 7.15; p greater than .10). ROC analysis also demonstrated excellent predictor performance (area index = 0.92 +/- 0.02). PMID- 3048901 TI - Low dose ibuprofen reverses the hemodynamic alterations of canine endotoxin shock. AB - The dose response of the nonsteroidal anti-inflammatory drug, ibuprofen, was evaluated in order to determine the most efficacious dose in the treatment of canine endotoxin shock. Fifteen minutes after an infusion of Escherichia coli endotoxin, four groups of dogs were given a single iv dose of 1, 5, 10, or 20 mg/kg of ibuprofen. These groups were compared to endotoxin only and saline control groups. All ibuprofen doses significantly improved the systolic, diastolic, and mean systemic arterial BP. The improvement in systemic BP was accompanied by an increase in the systemic vascular resistance. Pulmonary vascular pressures and resistance also increased after ibuprofen administration. The lack of a dose response and the demonstrated beneficial effect of low dose ibuprofen in the reversal of the hypotension associated with experimental canine endotoxin shock lead us to recommend the use of low dose ibuprofen for future endotoxin and sepsis studies. Use of low dose ibuprofen might have less of an effect on renal perfusion and would therefore be more likely to produce the beneficial hemodynamic response without compromising renal function. PMID- 3048902 TI - Portal thrombosis complicating appendicitis: ultrasound detection and hepatic computed tomography lobar attenuation alteration. AB - Lobar attenuation difference of liver on computed tomography was seen in a case of portal pyemia complicating perforated appendicitis. Left portal vein thrombus was detected first by ultrasound and subsequently confirmed by computed tomography. The left lobe of the liver showed greater contrast enhancement during the dynamic computed tomography study. The possible cause of this lobar attenuation difference is discussed. PMID- 3048903 TI - Neutropenic colitis: evaluation with computed tomography. AB - The computed tomography findings of 10 patients with neutropenic colitis are described and illustrated. Seven of these patients had leukemia, one had lymphocytic lymphoma, and two had systemic lupus erythematosus. All patients had colon wall thickening which was either isodense with the normal bowel tissue or showed areas of intramural low density. Air in the thickened bowel wall was seen in six patients. These computed tomography findings in neutropenic patients with fever, abdominal pain, and diarrhea should suggest the diagnosis in most instances, resulting in prompt treatment of this usually life-threatening entity. PMID- 3048905 TI - Ambulatory electrocardiographic recordings: the Holter monitor. PMID- 3048904 TI - Abdominal cryptococcoma in AIDS: a case report. AB - Cryptococcosis is a recognized opportunistic pathogen in the acquired immune deficiency syndrome. Although central nervous system infection and disseminated cryptococcosis is common in acquired immune deficiency syndrome, localized infection is rare. We present a case of massive retroperitoneal and mesenteric adenopathy in a male homosexual patient with acquired immune deficiency syndrome with clinical and radiologic features suggestive of lymphoma. However, this was proven pathologically to represent cryptococcal infiltration of the lymph nodes. Our experience indicates that Cryptococcus neoformans should be included in the differential diagnosis of massive abdominal adenopathy in the acquired immune deficiency syndrome. PMID- 3048906 TI - Hypertension in children. PMID- 3048909 TI - The desmoid (Reitamo) syndrome: etiology, manifestations, pathogenesis, and treatment. PMID- 3048908 TI - Cardiac nuclear medicine. AB - In this paper, the principal applications of nuclear medicine to studies of the heart are described. First, gated cardiac blood pool imaging is discussed, then thallium-201 myocardial imaging, myocardial infarct scintigraphy with 99mTc pyrophosphate, and evaluation of intracardiac shunts. In gated cardiac blood pool imaging, the patient's red blood cells are labeled with 99mTc. Images of the cardiac blood pool are then obtained in multiple projections and displayed in an endless-loop cine display. Quantitative indices of cardiac function are readily obtained, and a variety of functional images can be generated. Blood pool imaging may also be performed with use of a first-pass technique that yields similar information. Applications of blood pool imaging are discussed. The theory and techniques of planar and tomographic thallium-201 myocardial imaging are described, together with their application in the diagnosis of coronary artery disease. The prognostic value of thallium imaging is also examined. Myocardial infarct imaging with 99mTc pyrophosphate is described, and clinical indications are reviewed. Left-to-right cardiac shunts can be evaluated by following the first transit of a bolus of radiopharmaceutical through the lungs. Right-to-left shunts may be evaluated by injection of 99mTc macroaggregated albumin. PMID- 3048907 TI - Adolescent development: a biopsychosocial review. PMID- 3048910 TI - Surgery and current management for cancer of the esophagus and cardia: Part I. PMID- 3048911 TI - Cutaneous mucormycosis in a heart transplant patient. AB - We present the first case of cutaneous mucormycosis reported in a patient who had undergone a heart transplant operation. This appeared to be a localized infection without a predisposing local factor such as a surgical adhesive and without evidence of dissemination. Prompt treatment with intravenous amphotericin B resulted in an apparent cure. PMID- 3048912 TI - Delayed reaction to "lead" pencil simulating melanoma. AB - A 74-year-old man had sustained a penetrating lead pencil injury to his right lower leg at the age of fifteen. A dark gray macule occurred that remained unchanged for fifty-eight years. An enlarging black nodule appeared in this area, simulating a melanoma, when the patient was seventy-three years old. Histologic examination revealed exogenous pigment and a foreign body reaction. PMID- 3048913 TI - Subcorneal pustular dermatosis with seronegative polyarthritis. AB - A 55-year-old woman presented with acute onset of subcorneal pustular dermatosis and a seronegative polyarthritis. There have been a few reports of subcorneal pustular dermatosis associated with arthritis. PMID- 3048915 TI - Acute rotavirus gastroenteritis in children. Clinical, epidemiological and immunological aspects. PMID- 3048916 TI - Neutrophilic granulocyte function. Quantitative leukocyte mobilization and function of circulating and exudative neutrophils. PMID- 3048914 TI - Naftifine cream in the treatment of cutaneous candidiasis. AB - Naftifine, a member of a new class of synthetic antifungal drugs, the allylamines, was evaluated for the treatment of cutaneous candidiasis. In a double-blind, parallel-group clinical trial, sixty patients with cutaneous candidiasis were randomly assigned to receive either naftifine cream 1 percent or its vehicle twice a day for three weeks. Two weeks after the end of therapy, 77 percent of the naftifine-treated patients were mycologically cured (negative results on potassium hydroxide preparations and culture) and had no clinically apparent disease, compared with 3 percent of the patients treated with vehicle (p less than 0.001). Side effects reported with naftifine cream were few and minor. PMID- 3048917 TI - The human pharmacology of platelet inhibitory drugs. PMID- 3048918 TI - Liver transplantation in a 48-year-old female with primary biliary cirrhosis. AB - In 1983, a Danish female with primary biliary cirrhosis underwent orthotopic liver transplantation (OLT). The transplantation took place in Groningen under a Danish-Dutch cooperation, at this writing, and more than four years after transplantation, the patient is still alive. The quality of her life has been dramatically improved, making possible her return to work. Liver tests, liver function, and biopsies are normal and without signs of development of primary biliary cirrhosis in the transplanted liver. Liver transplantation is now frequently performed at several centres throughout the world with an increasing success rate. In Scandinavia, centres have been established in Norway, Finland, and Sweden, whereas in Denmark the organisation of a programme for liver transplantation is still being discussed. Because of the extremely high costs, liver transplantation has to be approved medically and politically, and the brain death criteria have to be accepted before it can be considered as a generally accepted modality of treatment. PMID- 3048919 TI - The regulation of subcutaneous adipose tissue blood flow in the ischaemic forefoot during 24 hours. Studies using the 133-xenon wash-out technique continuously over 24 hours. AB - A method for continuous measurement of subcutaneous adipose tissue blood flow in the forefoot during 24 hours (SBF) is described. The method is based on the radioisotope wash-out principle using 133-Xenon. A portable semiconductor detector is placed just above a local depot of 1-2 microCi 133-Xenon in 0.1 ml isotonic saline injected into the subcutaneous adipose tissue in the forefoot. The detector is connected to a memory unit allowing for storage of data. Due to the short distance, the recorded elimination rate constant must be corrected for combined convection and diffusion of the radioactive indicator. Characteristic 24 hour blood flow patterns were unveiled in patients with normal peripheral circulation and in patients having ischaemic nocturnal rest pain. In normals SBF doubled from day to night. This is ascribed to the local veno-arteriolar sympathetic axon reflex, which induces arteriolar vasoconstriction when the transmural pressure of the veins exceeds approximately 25 mmHg. In patients having ischaemic rest pains SBF was reduced by 37% on the average from day to night. This was caused by nocturnal hypotension, which is reflected proportionally in the foot. As the resistance vessels most probably are fully dilatated in feet with rest pain, the blood pressure drop during sleep causes the perfusion pressure and thus blood flow to come below a certain critical limit. There was a pronounced correlation between the reduction of systemic mean arterial blood pressure and SBF. The patients complaining of intermittent claudication, but no rest pains showed a variety of changes in SBF compatible with the continuous spectrum of the peripheral arteriosclerotic disease. After reconstructive vascular surgery, the 24-hour blood flow pattern normalized although the ankle/arm systolic blood pressure index did not come within normal range. SBF during day-time activities decreased by up to 50% postoperatively. This is caused by the reappearance of the local, sympathetic, veno-arteriolar vasoconstrictor response. During sleep SBF increased by 71%. The term postreconstructive hyperaemia seems improper, at least in a long-term context, normalization of preoperative ischaemia is a more correct notation. The coefficient of variation of nocturnal SBF was calculated to 10%. The method thus seems apt as a monitor in medical therapy for occlusive arterial disease. Changes of lambda has, however, to be considered in each study. PMID- 3048920 TI - Blood flow rate, temperature, oxygen tension and consumption in the skin of adults measured by a heated microcathode oxygen electrode. AB - Reliable transcutaneous measurements of arterial oxygen tension are based on a maximum skin blood flow rate which is created by heating the skin, typically at an electrode temperature of 44 to 45 degrees C. This increase in skin blood flow rate creates an arterialization of the oxygen tension in the capillaries and the surrounding tissue. The heat conducted to the skin surface is removed by a combination of convection (skin perfusion) and conduction to the deeper layers of the skin. This heat transport to and through the skin surface causes a measurable temperature profile from the electrode surface to the capillary layer. By a blood flow cessation it is possible to change the temperature profile because the convective part of the heat consumption is eliminated and the conductive part can then be measured and subtracted. Using the forearm as measuring area and a heated tc-PO2 electrode several observations were made. The mean temperature gradient over epidermis down to the capillary layer at an electrode temperature of 43, 44, and 45 degrees C was 2.1, 2.4 and 2.7 degrees C, respectively. The change in temperature profile caused by the blood flow cessation enabled primarily an estimation of the skin blood flow rate by temperature measurements, ranging from 0.07 to 0.24 ml.cm-2.min-1. Increasing blood flow rates correlated to increasing tc-PO2 values. By means of a dynamically, thermally shielded tc-PO2 electrode it was possible to determine the skin blood flow rates in the same arbitrary units computed on the basis of the heat dissipation to the skin surface. Furthermore, it was possible to correlate these blood flow estimations to the cutaneous blood flow rates measured by 133Xe washout technique. By increasing the electrode temperature the cutaneous blood flow rates increased from 12 to 50 ml.(100 g) 1.min-1. It was possible to calculate a conversion factor on the basis of the correlation between the heat determinations of the skin blood flow rate and the 133Xe measurements. Using this conversion factor the highest blood flow rate did not exceed 55 ml.(100 g)-1.min-1. The subcutaneous blood flow rate increased accordingly with increasing electrode temperature. It was concluded that the measured heat consumption of the skin is effected by the heat removing capacity of the cutaneous as well as the subcutaneous blood flow. The cutaneous blood flow, however, was considered predominant in the transport of heat from the skin surface. By 50 times of stripping the skin surface, the cornified epidermal membrane was removed. This procedure increased the tc-PO2 values by on an average 3.6 kPa (27.1 mmHg).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3048921 TI - The diabetic arteriole: the impact of diabetic microangiopathy on microcirculatory control. PMID- 3048922 TI - Mechanical strength of trabecular bone at the knee. AB - Interest in the biomechanical properties of trabecular bone has expanded in response to the problems related to total and partial joint replacement with the knee joint constituting a main focus of attention. This relatively recent development has left a number of fundamental problems unanswered, especially related to the machining, storage and testing of trabecular bone specimens. Nevertheless, these studies have contributed to the understanding of the mechanical function of trabecular bone. Regarding the role of trabecular bone at the knee joint, the following conclusions may be emphasized (conclusions drawn from the author's previous studies (I-X) are shown in italics): (1) Trabecular bone is almost exclusively responsible for the transmission of load at the proximal tibial epiphysis from the knee joint to the metaphysis. The peripheral shell surrounding the epiphysis is not composed of cortical bone and plays a negligible role in load transmission. (2) The compressive strength and stiffness of trabecular bone is primarily dependent upon the apparent density, trabecular architecture and the strength of the bone material. Direct and indirect sources suggest that the true material strength of trabecular bone is less than that of cortical bone. The epiphyseal trabecular architecture, featuring a marked polarity with alignment of primary trabeculae at right angles to the joint surface, is responsible for functional anisotropy which points to the axial compressive properties as the more important mechanical parameters. (3) Tensile and shear properties are of special relevance to mechanical loosening of implants. These properties may be derived from the apparent density, and a close empirical relation to the axial compressive strength and stiffness is suggested. (4) The foam-like structure of trabecular bone is the basis for the large energy absorptive capacity. (5) The pattern of axial compressive stiffness and strength at the normal proximal tibia differs little among individuals. Supporting the medial tibial plateau is a large high strength area with maximal strength centrally and slightly anteriorly, while laterally there is a restricted area of relatively high strength posteriorly with a lower maximal value than medially. Bone strength is significantly reduced within ten millimeters of the subchondral bone plate, and this reduction continues distally at the lateral condyle. At both condyles strength is reduced towards the periphery with very low values being obtained at the margins of the condyles and at the intercondylar region. Absolute bone strength values are influenced by the level of physical activity.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3048923 TI - Experimental metastasis. Dissemination and growth of lymphoma cells in mice. PMID- 3048924 TI - Contributions to the taxonomy of the genus Flavobacterium. PMID- 3048925 TI - Triple therapy and initial renal graft perfusion. AB - Cyclosporin A may decrease the perfusion of transplanted kidneys, especially if they are ischaemic. Therefore, monitoring of graft perfusion was performed with intravenous 99mTc pertechnetate angiography in 25 patients within 36 hours after renal transplantation and repeated 24 or 48 hours later and then again 48 hours later. In all patients, immunosuppression was performed with triple therapy (low dose prednisone, azathioprine, and cyclosporin A) which included a delay in cyclosporin A administration to the second postoperative day. Treatment with triple therapy did not cause a decrease in renal blood flow in the days immediately after transplantation as single therapy with high-dose cyclosporin A has been reported to do. PMID- 3048926 TI - Postoperative radionuclide monitoring of renal allografts on triple therapy. AB - Renal allograft dysfunction arising from rejection or cyclosporin A nephrotoxicity in patients on high-dose cyclosporin A administration can currently only be distinguished reliably by allograft biopsy excluding rejection. The renographic course up to 35 days after transplantation or to discharge was followed in 30 patients treated with low-dose azathioprine, prednisone, and cyclosporin A (triple therapy). Routine renographic monitoring combined with intravenous angiography in critical periods proved a valuable aid in the differentiation between post-operative complications such as acute rejection and surgical complications. PMID- 3048927 TI - Flow resistance of expiratory positive-pressure systems. AB - We measured the flow-resistance of five commercially available 10 cm H2O expiratory positive-pressure (EPP) valves (n = five per valve type) at bias flows of between 0 and 2,000 ml/s. We found that individual valves of each type and manufacturer functioned similarly. Different valve types, however, functioned differently: with one type, system pressure was higher than rated (p less than 0.05), and with another type, system pressure was significantly flow-dependent (p less than 0.01). The remaining types of valves had no flow-resistive properties and maintained a system pressure of 10 cmH2O. We conclude that system pressure is not similar in all continuous positive airway pressure (CPAP) systems using bias flow and EPP valves. The work of breathing imposed by CPAP circuits will be increased in systems whose EPP valves have flow-dependent properties. PMID- 3048929 TI - The role of pyrazinamide in tuberculosis chemotherapy. AB - Pyrazinamide is an antituberculosis drug synthesized in the 1950s and formerly used only as salvage therapy. Recent developments have elevated it to a central role in tuberculosis chemotherapy as the essential addition to isoniazid and rifampin which makes it possible to successfully complete treatment in six months. This is accomplished with no increase in hepatotoxicity. The only substantial side effect of this drug given at the dosage and for the duration used in these six-month regimens is a polyarthralgia which is only bothersome and not sufficient to warrant interruption of therapy. More rarely, acute gout is produced. The early history and pharmacology of this now first line antituberculosis drug are reviewed herein. PMID- 3048928 TI - Immunologic monitoring of the cardiac transplant patient. AB - Peripheral blood lymphocyte morphology and cell surface markers were repeatedly evaluated in 49 orthotopic cardiac transplant patients over a period of three years to determine the utility of these parameters in predicting an episode of acute cardiac rejection. Lymphocytes were measured with a calibrated microscope and termed "activated" if greater than 10 mu in diameter. If a patient demonstrated greater than 30 activated lymphocytes/cu mm, the same lymphocytes were stained with monoclonal antibodies directed to T-cell subsets and B cells. These findings were retrospectively correlated with endomyocardial biopsy results. Absolute numbers of T-cell subsets and B cells were analyzed via Student's t test to identify significant differences during acute cardiac rejection. Of 347 biopsy specimens, 47 demonstrated histologic features of acute cardiac rejection. Simultaneous immune activation of lymphocytes occurred with 33 of 47 samples, while another 51 episodes of lymphocyte activation were detected without acute rejection. No statistically significant differences in absolute numbers of T-lymphocyte subsets were noted at the time of acute rejection. PMID- 3048930 TI - Bronchopleurobiliary fistula. A complication of intrahepatic biliary stent migration. AB - We describe the management of bronchopleurobiliary fistula in a 56-year-old woman who underwent a (L) mastectomy with postoperative radio- and chemotherapy for advanced breast carcinoma and required insertion of inhabiliary Silastic stents for the relief of severe obstructive jaundice. During restaging of her carcinoma for further chemotherapy, she complained of dyspnea, right chest pain and productive cough with yellow sputum. Her chest x-ray film and thoraco-abdominal CT scan demonstrated right pleural effusion with a stent protruding through the right hemidiaphragm. The objective evidence of bile in the pleural aspirate with history of bile-stained sputum established the diagnosis of bronchopleurobiliary fistula resulting from biliary stent migration. PMID- 3048931 TI - Resistance of expiratory positive pressure valves. PMID- 3048932 TI - [Evaluation of the biological compatibility of joint prostheses]. PMID- 3048933 TI - [Leg lengthening: the value of echography in the evaluation of various phases of bone regeneration]. PMID- 3048934 TI - [Cysts of the popliteal cavity: use of echographic examination]. PMID- 3048935 TI - Cefoxitin for one day vs. ampicillin and metronidazole for three days in elective colorectal surgery. A prospective, randomized, multicenter study. AB - In a multicenter study the prophylactic efficacy of two antibiotic regimens was tested against postoperative septic complications following elective colorectal surgery. The study was conducted in a prospective block-randomized design. Patients were preoperatively allocated to either ampicillin, 1 gm, four times daily, and metronidazole, 0.5 gm, three times daily, for 72 hours, or to cefoxitin, 2 gm, given three times in a period of 10 hours. Both regimens were initiated immediately before surgery. Forty-five patients were withdrawn from the study after randomization. Three hundred fifty two patients (175 receiving ampicillin and metronidazole and 177 receiving cefoxitin) completed the study and were followed for one month postoperatively. The frequency of septic and nonseptic complications was not statistically significant different between the two regimens. About one third of all septic complications appeared more than two weeks after surgery. It is concluded that short-term treatment with cefoxitin is at least as efficient as a three-day treatment with ampicillin and metronidazole. PMID- 3048936 TI - Rectal peptic ulceration--a rare cause of rectal bleeding. Report of a case. AB - Analysis of the 27 cases of heterotopic gastric mucosa reported in the literature and a new case described here elucidates the main features of this disease: 1) all but one asymptomatic case were diagnosed in infants or in adults under 26 years old; 2) although rectal bleeding occurred in 24 patients, rectal peptic ulceration was found in only 13; 3) six of the patients also had rectal duplication; and 4) 19 times the limited extension of the heterotopic gastric mucosa was compatible with a complete excision by a transanal approach. PMID- 3048937 TI - Benign cystic ovarian teratoma with colorectal involvement. Report of a case and review of the literature. AB - Benign cystic ovarian teratoma is not an uncommon tumor, but its presentation as a bleeding rectal polyp has not been reported previously. A case report and review of the English medical literature are presented with emphasis on the cause, clinical presentation, and management of benign cystic ovarian teratomas with colorectal involvement. PMID- 3048938 TI - Alan Guyatt Parks 1920-1982. Proctocolectomy without ileostomy for ulcerative colitis. By A.G. Parks, R.J. Nicholls, 1978. PMID- 3048939 TI - Endoscopic laser surgery in the colon and rectum. AB - The first applications of laser energy in the gastrointestinal tract occurred barely a decade ago. Since then, endoscopic laser therapy has become widespread in the management of colonic and rectal disease. Applications have been developed for the palliative therapy of obstructing or bleeding malignancies, for the management of some benign and premalignant mucosal diseases and for certain anatomic problems such as anastomotic strictures. Recurrent bleeding from mucosal vascular lesions can be controlled with laser therapy. Methods for treating anorectal disorders also are evolving. The continuing development of new endoscopic systems for increasing ease of access to the gastrointestinal tract all point to ever-increasing applications for laser therapy in colonic and rectal surgery. PMID- 3048940 TI - Chronic obstructive pulmonary disease. AB - Chronic obstructive pulmonary disease (COPD) is equated with chronic bronchitis and emphysema as one disease entity. In COPD airflow limitation is relatively persistent--unlike asthma. Tests for "small-airways disease" form no part of routine practice, for their accuracy in detecting pathological change is debatable. The proteolytic theory of the pathogenesis of emphysema highlights the role of neutrophil elastase, antielastases, oxidants, antioxidants, and thus of potential new treatments. Clinical features of COPD include breathlessness, cough, and sputum, with airflow obstruction and lung hyperinflation. The differential diagnosis includes bronchiectasis, cystic fibrosis, and pulmonary hypertension, but pulmonary fibrosis, etc., is distinguished by radiological infiltrates. Plain chest radiography cannot reliably diagnose emphysema in life, but a new method measuring lung density from the computed tomographic (CT) scan allows location, quantitation, and diagnosis of emphysema (defined by enlargement of distal air spaces) in humans in life. "Pink puffers" with breathlessness, hyperinflation, mild hypoxemia, and a low PCO2 are contrasted with "blue bloaters" with hypoxemia, secondary polycythemia, CO2 retention, and pulmonary hypertension and cor pulmonale. Antismoking measures are a major aim in management. A bronchodilator regimen combining a slow-release oral theophylline with an inhaled beta 2-agonist, ipratropium, and high-dose inhaled steroids is proposed because even modest improvement in obstruction can help these patients. In acute exacerbations with purulent sputum, antimicrobials against Streptococcus pneumoniae and Hemophilus influenzae are used with controlled oxygen therapy aiming to keep the arterial PO2 over 50 mm Hg without the pH falling below 7.25. Influenza prophylaxis is recommended, but pneumococcal vaccination remains debatable. Chronic under-nutrition in "emphysema" implies controlled trials of feeding regimens--but these remain to be assessed. Long-term oxygen therapy is the only treatment known to prolong life in blue bloaters, and oxygen concentrators and transtracheal oxygen delivery are discussed. Pulmonary vasodilators (e.g., beta 2-agonists, hydralazine, nifedipine, angiotensin converting enzyme [ACE] inhibitors, etc.) have not yet been proved to provide long-term reduction in pulmonary arterial pressure. Blue bloaters have severe nocturnal hypoxemia in rapid eye movement (REM) sleep that is corrected by oxygen or the investigational drug almitrine.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3048941 TI - Computerized perimetry: possibilities for individual adaptation and feedback. AB - Computerized perimetry is often poorly accepted by the tested subjects, presumably because of sparse feedback and lack of adaptation to individual capacity. Several remedies are suggested, including visual response feedback, active correction of erroneous responses, various fixation prompts, and continuous adaptation to current reaction time. Intuitively intelligible result displays are also desirable. A novel format representing threshold level by symbol size may meet this need. PMID- 3048943 TI - Electroretinograms in early fungal endophthalmitis. AB - To facilitate the early diagnosis of exogenous fungal endophthalmitis, we developed a rabbit model for Aspergillus fumigatus endophthalmitis. Six eyes of six New Zealand white rabbits were inoculated with forty spores of A. fumigatus. A control group of their six contralateral eyes received a similar but sterile inoculum. The rabbit eyes were evaluated with respect to clinical appearance and the electroretinogram. Clinically evident endophthalmitis developed in all six infected eyes at an average of five days after injection (range 3-7 days). Data samples each 48 hours showed transient b-wave amplitude elevations in three infected eyes. These were greater than two standard deviations above the mean pre injection values (p less than 0.05). These amplitude increases preceded the onset of ophthalmoscopically recognizable infection and were observed at 2 to 5 days after injection (mean = 4 days). B-wave amplitudes in the infected eyes fell below two standard deviations of pre-injection values (p less than 0.05) at an average of 5 days (range 3-7 days) after injection, a corresponding to the onset of clinically obvious infection. At an average of 7 days after injection, all the infected eyes exhibited unmeasurable electroretinogram waveforms and severe infection. Histopathologic study of infected eyes showed extensive fungal infiltration of retina and vitreous tissues. The electroretinogram may be helpful in the early diagnosis of fungal endophthalmitis. PMID- 3048945 TI - Comparison of automated perimetry and pattern visually evoked cortical potentials in optic neuritis. AB - Twenty-one eyes of thirteen patients suffering from optic neuritis were examined with the Octopus automated perimeter and pattern visually evoked cortical potentials. All eyes had visual acuity of 0.2 or better. Visual field examination was measured by program 31 or 33, which tests the central 30-degree fields in a 6 degree grid. Pattern evoked potentials were obtained by reverse checkerboard stimulation of 3 rev/sec. A prolonged P100 latency was related to at least one abnormal point in the central nine points of program 31. All seven eyes with one abnormal point in the center had a delayed peak latency. The patients with either the normal central nine points of program 31 or normal peak latency had visual acuity of 0.6 or better. The graphs of visual acuity by Octopus compared with pattern evoked potential were very similar. With program Delta, the sensitivity loss ratio, i.e., the ratio of the mean loss at eccentricity of less than 10 degrees over less than 30 degrees, was related to the peak latency in optic neuritis caused by multiple sclerosis. PMID- 3048946 TI - [A new approach to enhancing the effects of postradiation recovery in various animal species]. PMID- 3048942 TI - The development of scotopic retinal function in human infants. AB - Scotopic retinal function undergoes age-related changes early in human infancy. Electroretinographic psychophysical, and pupillographic responses have been used in the study of normal development. Various components of the electroretinographic responses index distal and proximal retinal function. Changes in pupillary diameter, measurable in infants under carefully selected conditions, represent rhodopsin regeneration in the infants under carefully selected conditions, represent rhodopsin regeneration in the photoreceptor outer segment. From psychophysical data, inferences can be drawn about scotopic retinal control of visual performance. These data constrain theories about the determinants of sensitivity, about the flow of signals from the distal, rhodopsin bearing, outer segments to the proximal retina, and about modulations of straight through flow by feedback or inhibitory circuits. The results indicate that the post natal development of human scotopic function is due mainly to reorganization of processes central to the photoreceptors. PMID- 3048944 TI - Oscillatory potentials as a marker for dopaminergic disease. AB - In an effort to find electrophysiologic correlates of dopaminergic disease in man, we measured oscillatory potentials of the electroretinogram in normal and unmedicated schizophrenic subjects, and in rabbits given D1 agonist or antagonist drugs. We found highly reproducible oscillatory potentials in twelve male schizophrenics that were indistinguishable from those in nine male normal volunteers. We also found little electroretinographic difference among four rabbits given intramuscular injections of SKF 38393 (a D1 agonist) and four control animals injected with saline. However, four animals given SCH 23390 (a D1 antagonist) showed diminution of the b-wave and selective suppression of the second oscillatory potential. This effect was more prominent in oscillatory potentials generated by 5-Hz flicker than by a single flash. These rabbit data suggest that further efforts at finding clinical correlations may be worthwhile, possibly with trial of flicker oscillatory potentials, and they support the concept that different oscillatory potentials have different generators. PMID- 3048947 TI - [The structure of nucleosomes containing the mutant H2B histone]. PMID- 3048948 TI - [2 new promoters for RNA-polymerase of the phage SP6 are located beyond the gene for RNA-polymerase]. PMID- 3048949 TI - [Selection of mutants with nondisjunction of minichromosomes in Saccharomyces cerevisiae]. PMID- 3048950 TI - [Limited proteolysis of diphtheria toxoids in the periplasm of Escherichia coli and Erwinia carotovora]. PMID- 3048951 TI - Use of extract of Ruscus aculeatus in venous disease in the lower limbs. AB - The effectiveness and tolerability of a venotropic drug (RAES) composed of an extract of Ruscus aculeatus (16.5 mg), hesperidin (75 mg) and ascorbic acid (50 mg) were evaluated in 40 patients (30 female, 10 male) aged between 28 and 74 years, suffering from chronic phlebopathy of the lower limbs. The cross-over, double-blind trial involved two periods of treatment of 2 months with the drug (2 capsules, 3 times/day) or with placebo, and an interim period of 15 days for wash out. An overall tendency for improvement occurred that was more distinct during the periods of treatment with the drug. In fact, symptoms and plethysmographic parameters (in particular MVIV 40 and 60) immediately changed significantly in correspondence with the administration of RAES. The biological and clinical tolerability were excellent. PMID- 3048952 TI - A long-term study on the use of evening primrose oil (Efamol) in atopic children. AB - The effect of essential fatty acids on atopic eczema is controversial. Some workers have reported that patients with atopic eczema improved following oral treatment with evening primrose oil (an oil with a high concentration of gamma linolenic acid), but others have disputed this. This study was designed to look at the effect of evening primrose oil as a long-term oral supplementation for children with atopic eczema. Treated children dramatically improved their clinical condition after 4 weeks of therapy, and this improvement was maintained during the whole period of treatment (20 weeks). At the same time, modifications in plasma, neutrophil and lymphocyte fatty acid composition were detected. PMID- 3048953 TI - Evening primrose oil (Efamol) in the treatment of children with atopic eczema. AB - It has been reported that essential fatty acid levels may be low and that there may be reduced levels of delta-6-desaturase metabolites of linoleic acid in patients with atopic eczema. Good therapeutic results have been reported on the use of evening primrose oil (Efamol) in adults but not in children. Efamol contains gamma-linolenic acid, the delta-6-desaturase metabolite of linoleic acid. The authors have studied 24 children with atopic eczema: 12 of them were treated with a higher dose of evening primrose oil than in previous studies and 12 with placebo olive oil. The clinical status and plasma, neutrophil and lymphocyte fatty acid composition in these children have been evaluated. After 4 weeks the eczema of essential fatty acid-treated children significantly improved in comparison with that of placebo-treated children (p less than 0.01). There were significant changes in plasma fatty acid composition between the basal values and the end of active treatment, and between the placebo and actively treated children. Neutrophil and lymphocyte fatty acid composition did not seem to be related to disease activity. PMID- 3048954 TI - Clonidine and the treatment of the opiate withdrawal syndrome. AB - Clonidine is a central alpha adrenergic agonist which can be used to treat the opiate withdrawal syndrome. It has been used in many controlled trials and a substantial body of research evidence is available about its effectiveness in this role. This paper reviews the literature regarding its introduction in the Yale studies, its effectiveness relative to gradual methadone reduction treatments, its side effects, and touches briefly upon its use in conjunction with opiate antagonists. It is concluded that clonidine produces marked reduction of withdrawal symptoms but does not eliminate them; that the pattern of withdrawal symptoms differs from that associated with methadone reduction schemes; that there is some disagreement about the clinical significance of hypotensive and other side effects; and that the drug has interesting possibilities for rapid withdrawal programmes when combined with naltrexone. PMID- 3048955 TI - A rational approach to the IUCD. PMID- 3048956 TI - [Cefotaxime-induced allergic agranulocytosis in an acute attack of serologically atypical primary biliary cirrhosis]. AB - A 35-year-old woman developed pharyngitis with high fever and painful joint swellings. A severe cholestatic hepatitis occurred 40 days later with a rise of bilirubin to 32 mg/dl. "Nuclear dot" antibodies were demonstrated in the immunofluorescence test on cell cultures, confirming a diagnosis of primary biliary cirrhosis which had followed an atypical course. After nine days of cefotaxime administration, commenced because of persistent fever of 40 degrees C, an agranulocytosis was demonstrated, which regressed within a week of discontinuing the drug. The allergic genesis of the agranulocytosis was proven by repeated lymphocyte stimulation tests in the presence of cefotaxime. The autoimmune hepatitis was probably a predisposing factor in the genesis of the allergically induced agranulocytosis. PMID- 3048957 TI - [Therapy of disk prolapse. Chemonucleolysis versus percutaneous nucleotomy]. PMID- 3048959 TI - [Suspected syphilis during pregnancy due to cross reactions in Borrelia infection]. AB - A weakly positive titre (1:20) in the Treponema pallidum haemagglutination test and a highly positive titre (1:1280) in the fluorescence Treponema antibody absorption test, but negative result for IgM antibodies, were found in the serum of a 23-year-old pregnant woman. The cardiolipin microflocculation test was at first borderline positive, but negative on repeat. In the absence of a history of syphilis tests for Borrelia antibodies were performed. Those for antibodies against B. burgdorferi were highly positive in the ELISA test (550 units), in the indirect Borrelia immunofluorescence test 1:1280 for IgG antibodies and 1:160 for IgM antibodies. In the Borrelia-specific indirect haemagglutination test, which measures both IgG and IgM antibodies, the titres were 1:640 to 1:1280. These results confirmed the presence of an infection with B. burgdorferi and not with Treponema pallidum. PMID- 3048958 TI - [Piezoelectric lithotripsy of gallstones. Initial clinical experiences]. AB - Piezoelectric lithotripsy was undertaken on 50 patients with gallbladder stones, none of them requiring anaesthesia, analgetics or sedatives. Stone fragmentation was achieved in all patients during the first treatment. In 44 patients the maximum fragment size was less than 50% of the initial stone diameter. The mean maximum fragment size after the first treatment was 4.3 mm (+/- 3.3 mm). After a follow-up of 0-2 months in 14 of the 50 patients and of 2-4 months in 6 of 13 patients, no more stones could be seen by ultrasonography. After an average period of 8 weeks, 17 of 50 patients were free of stones. Piezoelectric lithotripsy did not have any severe side effects besides a mild pancreatitis in one patient. PMID- 3048960 TI - [Functional relations between the liver and kidney. II. Water excretion and hepatorenal syndrome]. PMID- 3048962 TI - [Villous adenoma of the papilla of Vater and acute necrotizing pancreatitis]. AB - In two patients a villous adenoma of the papilla of Vater was found to be the cause of an acute necrotizing pancreatitis. The diagnosis was largely made by early gastroduodenoscopy and endoscopic retrograde cholangiopancreatography. After initially conservative treatment of the pancreatitis the adenoma was excised transduodenally in both patients. Histological examination in both revealed a completely removed benign villous adenoma. Both patients have remained free of symptoms after 18 and 6 months, respectively. These observations support the call for intensive diagnostic measures when a diagnosis of pancreatitis has been made: tumours of the pancreas and of the periampullar region should be ruled out. PMID- 3048961 TI - [Glomerular-capillary hypertension. Experimental findings and their clinical importance]. PMID- 3048963 TI - [Occult ectopic Cushing's syndrome]. PMID- 3048964 TI - [Opportunistic infections and tumors of the gastrointestinal tract as manifestations of AIDS]. PMID- 3048965 TI - [Exchange transfusion and (or) plasmapheresis in more severe falciparum malaria?]. PMID- 3048966 TI - [Assessment of fluid content of the lung by thoracic radiography. Comparison of extravascular lung water measurement and results of radiological estimation methods]. AB - Extravascular lung water (EVLW) was determined with the thermal-dye double indicator method (119 individual measurements) in 23 patients in an intensive care unit, and the results were compared with estimates made from largely standardized portable chest X-ray films, using the staging method of Sibbald as well as that of Halperin. There was a significant correlation with the measured EVLW for both methods. Radiologically "normal" films corresponded to a mean EVLW value of 8.4 ml/kg body-weight. Radiologically judged adjacent stages did not in all cases compare with corresponding measured EVLW values. But when EVLW values were clearly abnormal, the X-ray films always demonstrated massive interstitial or alveolar infiltrations. Measurement of EVLW enables one accurately to judge the fluid contents of the lung and is superior to the assessment of the chest X ray film. PMID- 3048967 TI - [Breast-preserving therapy of breast carcinoma]. PMID- 3048968 TI - [Pharmacological interactions during treatment with cytostatic agents]. PMID- 3048969 TI - [Domestic animals as excreters of Clostridium difficile]. PMID- 3048970 TI - The cadherins: cell-cell adhesion molecules controlling animal morphogenesis. AB - Cadherins are a family of glycoproteins involved in the Ca2+-dependent cell-cell adhesion mechanism which is detected in most kinds of tissues. Inhibition of the cadherin activity with antibodies induces dissociation of cell layers, indicating a fundamental importance of these molecules in maintaining the multicellular structure. Cadherins are divided into subclasses, including E-, N- and P cadherins. While all subclasses are similar in molecular weight, Ca2+- and protease-sensitivity, each subclass is characterized by a unique tissue distribution pattern and immunological specificity. Analysis of amino acid sequences deduced from cDNA encoding these molecules showed that they are integral membrane proteins of 723-748 amino acids long and share common sequences; similarity in the sequences between subclasses is in a range of 50-60% when compared within a single animal species. L cells, with very little endogenous cadherin activity, transfected with the cadherin cDNA acquired high cadherin-mediated aggregating activity. Their colony morphology was altered by the ectopic expression of cadherins from the dispersed type to the compact type, providing direct evidence for a key role of cadherins in cell-cell adhesion. It has been suggested that cadherins bind cells by their homophilic interactions at the extracellular domain and are associated with actin bundles at the cytoplasmic domain. It appears that each cadherin subclass has binding specificity and this molecular family is involved in selective cell-cell adhesion. In development, the expression of each cadherin subclass is spatiotemporally regulated and associated with a variety of morphogenetic events; e.g. the termination or initiation of expression of a cadherin subclass in a given cell collective is correlated with its segregation from or connection with other cell collectives. Antibodies to cadherins were shown to perturb the morphogenesis of some embryonic organs in vitro. These observations suggest that cadherins play a crucial role in construction of tissues and the whole animal body. PMID- 3048972 TI - Neural cell adhesion molecules during embryonic induction and development of the kidney. AB - The neural cell adhesion molecules (N-CAM) are a family of related glycoproteins with Mr of 180, 140 and 120 x 10(3) (180K etc.). In the embryo, they are often highly sialylated and migrate as a diffuse band of 170-250K. N-CAM are found in non-neural tissues and we have now studied the expression of N-CAM in the developing mouse kidney. During kidney development, a unique conversion of a mesenchyme to an epithelium occurs and it is thought that this is mediated by an increase in cell adhesivity. By immunofluorescence, we show that N-CAM is present already at onset of kidney development on the cells of the uninduced nephrogenic mesenchyme. After induction, when the cells convert into an epithelium, they lose N-CAM gradually and instead begin to express uvomorulin, another primary CAM. By using an organ culture model, we could rather precisely show that N-CAM and uvomorulin are coexpressed for a short period, but, when epithelial cell polarization is evident, only uvomorulin is present on the epithelium, whereas N CAM is confined to the surrounding mesenchyme. Immunoblotting for N-CAM revealed that the 'embryonic' form of N-CAM, the broad 170-250K band was not present in the embryonic kidney, which instead expressed the three distinct 180K, 140K and 120K bands typical of adult neurones. The 180K and 140K bands were gradually lost during development and were no longer detectable in adult kidneys. By using an N CAM cDNA, we detected three different mRNAs of 7.4, 6.7 and 4.3 kb in the developing kidney, but this expression was restricted to the embryonic and early postnatal stages. No transcripts were detectable in adult kidneys. The studies do not support the hypothesis that N-CAM expression in the kidney is turned on by embryonic induction. Rather, we suggest that N-CAM are important adhesives for the predetermined, but not yet induced, nephrogenic mesenchyme. PMID- 3048971 TI - Embryonic vascular development: immunohistochemical identification of the origin and subsequent morphogenesis of the major vessel primordia in quail embryos. AB - The development of the embryonic vasculature is examined here using a monoclonal antibody, QH-1, capable of labelling the presumptive endothelial cells of Japanese quail embryos. Antibody labelling is first seen within the embryo proper at the 1-somite stage. Scattered labelling of single cells appears ventral to the somites and at the lateral edges of the anterior intestinal portal. The dorsal aorta soon forms a continuous cord at the ventrolateral edge of the somites and continues into the head to fuse with the ventral aorta forming the first aortic arch by the 6-somite stage. The rudiments of the endocardium fuse at the midline above the anterior intestinal portal by the 3-somite stage and the ventral aorta extends craniad. Intersomitic arteries begin to sprout off of the dorsal aorta at the 7-somite stage. The posterior cardinal vein forms from single cells which segregate from somatic mesoderm at the 7-somite stage to form a loose plexus which moves mediad and wraps around the developing Wolffian duct in later stages. These studies suggest two modes of origin of embryonic blood vessels. The dorsal aortae and cardinal veins apparently arise in situ by the local segregation of presumptive endothelial cells from the mesoderm. The intersomitic arteries, vertebral arteries and cephalic vasculature arise by sprouts from these early vessel rudiments. There also seems to be some cell migration in the morphogenesis of endocardium, ventral aorta and aortic arches. The extent of presumptive endothelial migration in these cases, however, needs to be clarified by microsurgical intervention. PMID- 3048973 TI - Spinal opioid analgesia. A critical update. AB - Small spinal (intrathecal or extrathecal) doses of opioids induce a long-lasting and regional analgesic effect in various experimental animal models. Nowadays extrathecal morphine administration is considered an established method of controlling postoperative and cancer-induced pain conditions. The potency of morphine applied by the spinal route is higher than when the drug is applied by the intravenous (IV) route. Opioids which are more lipophilic than morphine will provide a marginally better analgesic effect when administered by the spinal route as compared with the IV route. Several controlled clinical trials in postoperative patients have demonstrated that a single dose of morphine administered by the spinal route gives a more long-lasting action than a similar IV dose. It is not known whether frequent patient-controlled administration of morphine may provide equally good analgesia without additional side effects. The use of spinal morphine in the treatment of cancer-related pain is based on clinical experience only. There are risks in replacing opioid administration by the oral or IV route with spinal opioids. Morphine should only be used in selected cases until the advantage of spinal opioid analgesia to control postoperative and cancer pain has been clearly defined in well-designed clinical studies. Spinal morphine dosages must be individualised according to the intensity of the nociceptive stimuli and should take into account intra individual variability in drug responses due to pharmacokinetic and pharmacodynamic factors. PMID- 3048975 TI - Bromperidol. A preliminary review of its pharmacodynamic and pharmacokinetic properties, and therapeutic efficacy in psychoses. AB - Bromperidol is a close structural analogue of haloperidol. Both agents possess similar pharmacodynamic properties which are consistent with central antidopaminergic activity. The available clinical data from non-comparative and comparative trials have shown that bromperidol possesses antipsychotic activity. Its overall efficacy was slightly greater than chlorpromazine and perphenazine, and similar to or slightly better than haloperidol in a small number of comparative studies. Bromperidol frequently displayed a faster onset of action than these comparative agents and there have been reports of bromperidol causing an activating effect. However, extrapyramidal side effects occurred with similar frequency and severity after bromperidol compared with the 3 reference drugs. Data from additional controlled clinical trials are required to confirm the comparative antipsychotic efficacy of bromperidol and to assess whether it possesses any advantages in certain sub-types of schizophrenia. PMID- 3048979 TI - [Photodynamic therapy]. PMID- 3048978 TI - [Endocrine changes induced by systemic disease]. PMID- 3048974 TI - Ceftriaxone. A reappraisal of its antibacterial activity and pharmacokinetic properties, and an update on its therapeutic use with particular reference to once-daily administration. AB - Since ceftriaxone was first reviewed in the Journal, further studies have confirmed its broad antibacterial spectrum in vitro and extended its clinical documentation in comparative studies with other widely used drugs in infections of the urinary and lower respiratory tract, meningitis in infants and children, uncomplicated gonorrhoea, perioperative prophylaxis in patients undergoing surgery, and in several other types of infection. As in earlier studies, which primarily used a twice-daily dosage regimen, few significant differences were found between therapeutic groups in comparative studies and results have demonstrated the efficacy of once-daily ceftriaxone in all but the most serious infections, such as sole antibiotic therapy in pseudomonal infections. Wider clinical experience has established that ceftriaxone is generally well tolerated. Thus, ceftriaxone now has a well-defined place as an appropriate alternative for the parenteral treatment of a variety of infections due to susceptible organisms, as well as for perioperative prophylaxis of surgery, and may offer advantages of greater convenience over other parenteral antibiotics which are administered more frequently. PMID- 3048977 TI - Sustained release theophylline preparations. Practical recommendations for prescribing and therapeutic drug monitoring. AB - Theophylline is an important antiasthmatic medication which has bronchodilator properties. With increased understanding of the relationships of serum theophylline concentration and effect, both adverse and beneficial, oral dosage forms were developed to provide consistent serum theophylline concentrations with the benefit of convenient dosage intervals for long term use. Since factors such as concurrent disease states, drug interactions and age have a profound effect on theophylline disposition, relatively sophisticated dosage guidelines have evolved. Theophylline is in fact a model drug for the application of pharmacokinetic principles to the individualization of a treatment regimen. The purpose of this discussion is to review the relationship of serum theophylline concentration and pharmacodynamic effect and the special properties of oral sustained release theophylline formulations, and to provide a practical approach to prescribing theophylline. Guidelines are provided on the use of serum theophylline concentrations to individualize the theophylline dose, with an analysis of available techniques to monitor theophylline. PMID- 3048980 TI - [Toxocariasis in children]. PMID- 3048983 TI - [Survival and transplantation]. PMID- 3048981 TI - [Origin of autoimmune mechanisms]. PMID- 3048976 TI - Mechanisms of action of aminoglutethimide as endocrine therapy of breast cancer. AB - During the last decade aminoglutethimide has been recognised as a valuable alternative in endocrine therapy for advanced breast cancer. Although some side effects do occur, most often these are initial effects which subside within a few weeks, and cessation of therapy is not usually indicated. Aminoglutethimide was originally introduced as an inhibitor of steroidogenesis in the adrenal cortex. It was soon recognised, however, that inhibition of the non-glandular aromatase, blocking the conversion of androgenic prohormones to oestrogens, was more important, resulting in decreased blood levels of oestrogens. In this review the role of aromatase inhibition as the only important aspect of the mechanism of action of aminoglutethimide is challenged. Evidence has accumulated during the last few years that aminoglutethimide is a most potent inducer of microsomal enzymes. In addition to the pharmacological implications this has (suggesting important interactions), it also points to the possibility that levels of oestrogens are decreased due to accelerated metabolism of these hormones. Based on new experimental data, and also clinical work with alternative aromatase inhibitors, it appears that the antitumour activity of aminoglutethimide may be due to both aromatase inhibition and accelerated metabolism of oestrogens. This seriously challenges the importance of aromatase inhibition alone as a strategy in endocrine therapy of breast cancer, and furthermore suggests that accelerated metabolism of key hormones is an alternative strategy to be explored. PMID- 3048982 TI - [Direct ultrasonic diagnosis of salivary gland neoplasms]. PMID- 3048985 TI - [A heart transplant patient during the first year]. PMID- 3048984 TI - [Soft tissue images of the legs]. PMID- 3048986 TI - [Clinical research on the autonomic nervous system]. PMID- 3048987 TI - [Cervical pregnancy]. PMID- 3048988 TI - [Neurological and neuropathological expressions of HIV infection]. PMID- 3048989 TI - [Ultrasonic confirmation of vesico-ureteral reflux]. PMID- 3048990 TI - [Management of scrotal hydrocele by sclerotherapy]. PMID- 3048991 TI - [Venous thrombosis in spinal and spinal cord injuries]. PMID- 3048992 TI - [The physiology of sexual reactions in women]. PMID- 3048993 TI - [The physiology of male sexual activity]. PMID- 3048994 TI - [The endocrinology of sexual disturbances in the male]. PMID- 3048996 TI - [Homosexuality activity--a disturbance or a specific activity]. PMID- 3048995 TI - [Old age and sexual activity]. PMID- 3048997 TI - [Pharmacological causes of disturbances in sexual activity]. PMID- 3048998 TI - [Gynecologic problems and sexology]. PMID- 3048999 TI - [Brief psychotherapy for disturbances in sexual activity]. PMID- 3049000 TI - [Brief psychotherapy for sexual problems]. PMID- 3049002 TI - [Brain death]. PMID- 3049003 TI - [Growth factors and post-injury tissue repair]. PMID- 3049005 TI - [Death induced by verapamil overdose]. PMID- 3049004 TI - [Spontaneous dissection of the internal carotid artery]. PMID- 3049001 TI - [Urological contingencies in the management of sexual dysfunction in men]. PMID- 3049006 TI - [Radiology of multiple thoracic trauma]. PMID- 3049007 TI - [Life beneath the microscope]. PMID- 3049008 TI - [Gonadal differentiation]. PMID- 3049009 TI - [The stages of embryonic growth]. PMID- 3049010 TI - [Diagnosis and prognosis of fetal malformation]. PMID- 3049011 TI - [The fetal thyroid gland]. PMID- 3049012 TI - [The diabetic fetus]. PMID- 3049013 TI - [Prognosis and management of fetal growth]. PMID- 3049014 TI - [Fetal asphyxia]. PMID- 3049016 TI - [Fetal hydrops]. PMID- 3049015 TI - [Fetal rhythm disturbances]. PMID- 3049017 TI - [Medical treatment of the fetus]. PMID- 3049018 TI - [Congenital adrenal hyperplasia]. PMID- 3049019 TI - [Fluoroquinolones]. PMID- 3049020 TI - [Bone marrow transplantation--a promising new treatment for myeloma]. PMID- 3049021 TI - [Recording of sleep stages in depressed patients and control in the sleep laboratory]. PMID- 3049022 TI - [Is DNA aneuploidy a marker of malignant growth?]. PMID- 3049023 TI - [The Crick-Mitchison theory of sleep and psychoanalysis]. PMID- 3049026 TI - [Primary tubercular infection and BCG vaccination]. PMID- 3049024 TI - [Characteristics of coronary artery disease in the elderly]. PMID- 3049025 TI - [Erythema nodosum]. PMID- 3049027 TI - [Cleanliness and hygiene in the Finnish sauna]. PMID- 3049028 TI - [The universality of the sauna]. PMID- 3049029 TI - [Blood circulation in the sauna]. PMID- 3049030 TI - [Fluid balance and the sauna]. PMID- 3049031 TI - [Hormonal effects of sauna bathing]. PMID- 3049033 TI - [Pulmonary effects of steam]. PMID- 3049032 TI - [The sauna and the heart]. PMID- 3049034 TI - [Skin and venereal diseases and the sauna]. PMID- 3049035 TI - [The sauna and pregnancy]. PMID- 3049036 TI - [The sauna and alcohol]. PMID- 3049037 TI - [The sauna and sports]. PMID- 3049038 TI - [Interleukin 2, killer cells and cancer]. PMID- 3049039 TI - [Continuous indirect measurement of blood pressure]. PMID- 3049040 TI - [Medical consumption of over the counter drugs]. PMID- 3049041 TI - [Insulin pump therapy]. PMID- 3049042 TI - [Medicine in 1888 from Duodecim articles]. PMID- 3049043 TI - Prospects and problems for malaria vaccination: an African perspective. PMID- 3049044 TI - Demographic and socio-economic patterns among new admissions to Mathari Mental Hospital, Nairobi. PMID- 3049046 TI - [Pirenzepine and cimetidine in the treatment of duodenal ulcer]. AB - The efficacy of pirenzepin--of anticholinergic effect--and the H2-receptor blocking cimetidine has been studied in duodenal ulcer with random, double blind fashion. Recurrence examinations were carried out 6 and 12 months following the treatment. 50 patients were given pirenzepin and 50 cimetidine. The average age of the patients was 44.5 and 43.8 years respectively. Of them 60 (24 + 36) were men and 40 (26 + 14) women. At the start, 6 weeks following the start 6 and 12 months after the finishing of the treatment gastroscopy was performed. In the course of the six-week-long treatment 38 (76%) of the pirenzepin taking patients and 36 (72%) of the cimetidine taking patients recovered. No significant difference was found between the efficacy of the both treatments. In the respect of the half and one year recurrence, no significant difference was observed between the two patient groups. Ten of the patients taking pirenzepin and 4 of those taking cimetidine complained of side effects. Dryness of mouth, visual disturbance in the former group and constipation in the latter one. On the basis of the examinations both secretion inhibitors were found equally suitable for the therapy of duodenal ulcer. PMID- 3049045 TI - Terrestrial model food chain and environmental chemicals. I. Transfer of sodium [14C]pentachlorophenate between springtails and carabids. AB - A model soil food chain of a ruderal ecosystem has been constructed in order to study the uptake, transfer, and accumulation of [14C]pentachlorophenate (PCP-Na). The model was based on three food levels, viz. baker's yeast, collembola, and carabid beetles, and the contaminant chemical introduced was via initial food. Continuous exposure of the organisms to the test chemical resulted in a significant uptake and transfer of radiocarbon into the food chain elements. Bioaccumulation of radiocarbon in the body tissues of the organisms was low, as large amounts taken up were quickly eliminated through the excrements. The radiocarbon level of prey animals was about 100 times higher than that of their predators, but there was only small difference in concentration between collembolas and yeast. This was probably because of a faster excretion of the chemical by the beetles than by the collembolas. During the test period no conversion of [14C]PCP-Na took place in the yeast, but the collembolas and beetles metabolized 50 and 59%, respectively. Criteria are proposed for successful implementation of food chain models. PMID- 3049047 TI - [Intermediate filaments of the cytoskeleton--cell-specific markers in pathology]. PMID- 3049048 TI - Loss of nitrogen from organs in rats induced by exogenous glucagon. AB - Rats weighing 220 g were injected sc with zinc protamin glucagon 20 micrograms once daily (recurrent hyperglucagonemia) and zinc protamin glucagon 60 micrograms three times daily (chronic hyperglucagonemia); the controls received the vehicle three times daily. In the first group blood glucagon rose to above 200 ng/liter for 5 h every day; in the second group it constantly stayed above 600 ng/liter. After both 2 (n = 5) and 14 (n = 5) days treatment the control total blood alpha amino-nitrogen (AAN) concentration was 4.3 +/- 0.1 mmol/liter, and the urea nitrogen synthesis rate was 4.9 +/- 0.4 mumol/(min.100 g BW) (mean +/- SEM) in controls. In recurrent hyperglucagonemic rats, treated for both 2 (n = 5) and 14 (n = 5) days, total AAN was 3.6 +/- 0.2 mmol/liter (P less than 0.05 vs. control) and urea nitrogen synthesis rate 4.5 +/- 0.8 mumol/(min.100 g BW). In chronic hyperglucagonemic, treated for both 2 (n = 5) and 14 (n = 5) days, total AAN was 2.2 +/- 0.1 mmol/liter (P less than 0.05 vs. control) and UNSR 7.9 +/- 0.8 mumol/(min.100g BW) (P less than 0.05 vs. control). The urea excretion was identical in controls and during recurrent hyperglucagonemia, but it was increased by 50% during chronic hyperglucagonemia. Food intake was the same in all groups. N Balances decreased from 10 mmol/24 h to 5 mmol/24 h (P less than 0.05) by chronic hyperglucagonemia. The total organ N content did not change by recurrent hyperglucagonemia, but in chronic hyperglucagonemia it decreased to 65 85% (P less than 0.01) in carcass, intestines, liver, and kidneys. In conclusion chronic but not recurrent hyperglucagonemia increases the rate of urea synthesis and decreases the blood amino acid concentration. This is suggested to be a reason for the loss of N from organs by chronic hyperglucagonemia. PMID- 3049049 TI - Thyroid hormone regulation of alpha-lactalbumin: differential glycosylation and messenger ribonucleic acid synthesis in mouse mammary glands. AB - Mouse mammary tissue, when cultured in the presence of insulin, corticoids, PRL, and physiological levels of T3, shows increased synthesis and secretion of alpha lactalbumin. Tissue cultured in the presence of insulin, hydrocortisone, PRL, and T3 synthesizes two distinct forms of alpha-lactalbumin, but secretes only one form. Tissue cultured in the absence of T3 synthesizes and secretes only one form. To address the question of whether these two electrophoretically distinct forms arose by differential glycosylation of the same polypeptide or by synthesis of two different polypeptide precursor chains, mammary tissue was cultured in the presence of insulin, corticoids, and PRL with or without T3, and the mRNA and alpha-lactalbumins were isolated. Northern blot analyses indicated that mammary gland tissue cultured in the presence of T3 contained 2.46 times more alpha lactalbumin mRNA than tissue cultured only in the presence of insulin, hydrocortisone, and PRL. This enhanced mRNA level was confirmed by in vitro translation experiments where tissue cultured in the presence of insulin, hydrocortisone, PRL, and T3 produced mRNA that resulted in 2.1 times as much radiolabeled alpha-lactalbumin as tissue cultured in the absence of T3. Sodium dodecyl sulfate-polyacrylamide gel analysis of the in vitro translation products revealed only one band, suggesting the presence of only one message. Endoglycosidase digestion of the two forms of alpha-lactalbumin produced in the presence of T3 resolved them into a single band on sodium dodecyl sulfate polyacrylamide gels. Thus, the electrophoretic differences between the two forms synthesized in the presence of T3 appear to be due to differential N-linked glycosylation of the same polypeptide chain and not to synthesis of two different polypeptide precursor chains. PMID- 3049050 TI - Insulin and insulin-like growth factor I binding to cultured rat glomerular mesangial cells. AB - The potential effects of insulin and insulin-like growth factor I (IGF-I) on mesangial cell (MC) metabolism and growth were examined. Radiolabeled insulin or IGF-I were incubated with cell membranes from rapidly proliferating (subconfluent) or nonproliferating (confluent) MC in the presence of increasing concentrations of unlabeled heterologous and homologous ligands (0-10(-6) M). Insulin binding to MC was specific and saturable, with Scatchard analysis of binding data showing the characteristic curvilinear plot. The predicted insulin binding maximum of 4.2 X 10(-12) M/100 micrograms protein for a theoretical high affinity site was consistent with a relatively low density of receptors, which were the same in proliferating and nonproliferating cell preparations. Specific binding of IGF-I to MC was also demonstrated. Binding data for membranes from proliferating cultures generated a linear Scatchard plot, which predicted a binding maximum of 3.5-9.7 X 10(-11) M/100 micrograms protein and a Kd of 2.0-3.2 X 10(-9) M. In contrast, membranes from nonproliferating cultures had no demonstrable specific binding of IGF-I. Covalent cross-linking of radiolabeled IGF-I to membranes from subconfluent cells demonstrated specific binding to a 145K membrane protein. A 95K membrane protein from a partially purified receptor preparation demonstrated autophosphorylation when incubated with 5 X 10(-9) M IGF I. Incubation of MC with 10(-9) M IGF-I doubled cellular growth rates, an effect that could be duplicated only with high concentrations (10(-6) M) of insulin. These observations indicate that MC express predominantly receptors for IGF-I, and that growth stimulatory effects of physiological concentrations of IGF-I and pharmacological concentrations of insulin are probably mediated through the IGF-I receptor. PMID- 3049053 TI - Gastric involvement with excavated plasmacytoma: case report and review of endoscopic criteria. PMID- 3049054 TI - Physiology of the human sphincter of Oddi. AB - During the last 25 years relevant data which contribute to the clarification of pancreatico-biliary sphincter function, have been derived from physiological and pharmacological studies, and from cinefluorographic and manometric observations in man. The human sphincter of Oddi appears to be a complex device, relatively independent of the duodenum and endowed with a primarily occluding action. Perendoscopic, pre- and post-operative sphincter of Oddi manometry has opened up new prospectives for a better definition of old concepts, such as biliary dyskinesia and post-cholecystectomy syndrome. In addition, manometric measurement may provide a more rational basis for sphincterotomy, and could allow endoscopists and surgeons to objectively evaluate the extent of sphincter splitting. PMID- 3049052 TI - Calcitonin gene-related peptide stimulates adenylate cyclase activation via a guanine nucleotide-dependent process in rat liver plasma membranes. AB - To evaluate the functional relationship between the liver calcitonin gene-related peptide (CGRP) receptor and guanine nucleotide-binding proteins, we investigated the effects of nucleotides not only on adenylate cyclase activation by CGRP, but also on 125I-[Tyr0]rat CGRP binding to rat liver plasma membranes. In the presence of GTP, rat CGRP stimulated adenylate cyclase activity in a dose dependent manner in rat liver plasma membranes, and this effect was reduced in the absence of GTP. Salmon calcitonin also enhanced adenylate cyclase activation in the presence of GTP, but only in higher concentrations. On the other hand, guanine nucleotides not only decreased 125I-[Tyr0]rat CGRP binding to rat liver plasma membranes, but also accelerated the dissociation of label binding, and the removal of Mg2+ from incubation medium attenuated this inhibitory action of GTP on 125I-[Tyr0]rat CGRP binding to membranes. Scatchard analysis of the data revealed that the reduction of 125I-[Tyr0]rat CGRP binding by GTP was due to the decrease in binding affinity without a significant change in binding capacity. These findings lead us to conclude that binding of CGRP to its receptors activates adenylate cyclase in rat liver plasma membranes via a guanine nucleotide-dependent process, suggesting the involvement of guanine nucleotide binding stimulatory protein in the action of CGRP. PMID- 3049056 TI - Sphincter of Oddi manometry: comparison of microtransducer and perfusion methods. AB - There are two approaches to endoscopic sphincter of Oddi manometry, the microtransducer method and the perfusion method. Data reported by us and others are reviewed, with the aim of comparing the two techniques. Biliary pressure, pancreatic duct pressure, and sphincter of Oddi phasic wave activity can be measured by both of these methods. Easy handling and the capability of recording the mode of phasic wave propagation with a multilumen catheter are present advantages of perfusion manometry over the microtransducer method. Because there is no need for perfusion of fluid, the equipment needed for microtransducer manometry is simpler than that needed for the perfusion method, and the microtransducer technique may be more suitable for prolonged recording. An inter study comparison of manometric data is easier with microtransducer manometry, but must await the collection of more data. Manometric differences between various diseases are still inconsistent, whereas there is no question of the effect of sphincterotomy on pressure values, with the exception of pancreatic duct and sphincter pressures. The possibilities of diagnosing sphincter of Oddi dysfunction, and of predicting the response to sphincterotomy have been received with enthusiasm. However, whether the findings obtained by endoscopic manometry over relatively short periods of time can be considered representative of overall sphincter function is now being questioned since the presence of physiologic cyclic changes in sphincter of Oddi phasic activity in phase with the migrating motor complex of the duodenum was demonstrated. Hormonal or pharmacological stimulation of sphincter activity may help us overcome this problem. PMID- 3049055 TI - Pharmacology of the sphincter of Oddi. AB - The sphincter of Oddi is the smooth muscle connection between the bile duct and the duodenum. Its physiological function is associated with a regular motility characterized by phasic contractions superimposed on the sphincter of Oddi baseline pressure. Recently introduced ERCP-manometry permits further studies of sphincter of Oddi pharmacology. A number of drugs have so far been studied. Sedatives of the diazepam type had no effect on the sphincter, while butylscopolaminium bromide, a typical neurotropic agent, brings about cessation of the sphincter motility for 3-8 minutes. Hymecromon lowered the sphincter baseline pressure from 9.8 to 7.8 mmHg. A 1.2 mg sublingual dose of nitroglycerin, a typical musculotropic agent, caused significant relaxation of the sphincter, and decreased baseline pressure from 8.9 mmHg to 2.9 mmHg; Sphincter motility was not affected. Morphine-like analgetics, in particular pentazocine, elevated sphincter baseline pressure, but buprenorphine and tramadol did not. Pharmacological doses of gastrointestinal hormones also affect the sphincter; CCK octapeptide, glucagon and secretin are able to decrease sphincter of Oddi baseline pressure, and CCK octapeptide abolishes sphincter motility. Sphincter of Oddi pharmacology is of clinical interest. The administration of sphincter-relaxing agents, in particular nitroglycerin and butylscopolaminium bromide, enables the endoscopist to extract small common bile duct stones without previous papillotomy. Analgetics that induce sphincter contraction and thus hinder the flow of bile and pancreatic juice, may be helpful for the treatment of pain in patients with pancreatico-biliary disease. Investigations into the effect of CCK on the healthy and diseased sphincter permit us to identify patients with sphincter dysfunction using a special CCK-provocation test. PMID- 3049051 TI - Characterization of monoclonal antibodies specific to bovine renal vitamin D hydroxylases. AB - Monoclonal antibodies (MAbs) have been produced which recognize specific epitopes on bovine renal mitochondrial vitamin D3 1 alpha- and 24-hydroxylases. Renal mitochondria cytochrome P-450s were partially purified to 0.5-2 nmol/mg by Emulgen 911 and cholate solubilization, followed by chromatography on a 2-(4,6 dichloro-O-biphenyloxy)ethylamine HBR affinity column. Reduced carbon monoxide difference spectra determined that this preparation contained 0.5-2 nmol P-450/mg protein. This preparation contained both 1 alpha- and 24-hydroxylase activities, and Eadie-Hofstee plots of product formation as a function of substrate concentrations have maximum velocities of 1.4 and 4 pmol product/30 min.mg protein and Km values of 690 and 1300 nM, respectively. Bovine renal hydroxylases were isolated by immunoprecipitation from this partially purified P-450 preparation with a polyclonal antibody specific for rat liver microsomal cytochrome P-450 RLM5. This polyclonal antibody immunoprecipitated both 1 alpha- and 24-hydroxylase activities as well as renal mitochondrial cytochrome P-450, as determined by reduced CO spectra. Bovine renal mitochondrial components were immunoisolated and used to immunize BALB/c mouse spleen cells in vitro. MAbs then produced were screened for 1) immunoisolation of renal mitochondrial hydroxylase activity from a partially purified preparation, 2) immunohistochemical detection of antigen in renal proximal tubule cells, and 3) immunoquantitation of renal hydroxylases in a solid phase sandwich (enzyme-linked immunosorbent assay) and by 4) Western blot analysis. MAbs were isolated with specifically immunoprecipitated 1 alpha-hydroxylase activity, 24-hydroxylase activity, or both. In 10 micron sections of bovine kidney, antibodies detected antigen only in proximal tubule cells on the basal surface, which is rich in mitochondria. No antigen was detected in sections of pancreas or liver. In the solid phase sandwich enzyme linked immunosorbent assay, MAbs detected 1 alpha and 24-hydroxylases only in renal mitochondria and not in liver microsomes or adrenal gland mitochondria. In a Western blot, MAbs specific for epitopes expressed on both hydroxylases detected a single band(s) at 52,000-53,000 daltons. Apparently it is not possible to discriminate between hydroxylases by sodium dodecyl sulfate-polyacrylamide gel electrophoresis Western blots. By these criteria, MAbs have been generated which are specific to epitopes expressed on bovine renal mitochondrial 1 alpha- and 24 hydroxylases. PMID- 3049057 TI - Papillary stenosis. AB - The present state of papillary stenosis is reviewed. ERCP manometry has become the most important means of evaluating sphincter of Oddi dynamics. Pressure measurements in the sphincter segment appear useful to differentiate patients with sphincter of Oddi dysfunction from patients with an organic stenosis. The author's experience with sphincter of Oddi manometry, with endoscopic sphincterotomy and hydrostatic balloon dilation of the sphincter of Oddi, and the results of an international inquiry, are presented. PMID- 3049059 TI - Clinical and endoscopic aspects of tumors of the ampulla of Vater. AB - Diverse tumors may arise at the ampulla of Vater, although the clinical manifestations for all types of lesions are similar. Carcinoma and benign adenomas are the most important ampullary tumors, and there is evidence that adenomas are premalignant. Endoscopy offers the best prospect of accurate diagnosis, but the sensitivity and specificity of this diagnostic modality is less than expected. In particular, endoscopic biopsies may be falsely negative for carcinoma in a significant percentage of cases. When successful, ERCP may demonstrate dilated pancreatic and bile ducts and in some cases the presence of a tumor mass. The preferred treatment for virtually all ampullary tumors is surgery, the general types of operation being local resection, pancreatoduodenectomy, and biliary decompression procedures. The choice of operation depends on whether the tumor is benign or malignant, its extent, especially if malignant, and an assessment of the patient's ability to withstand radical resection. Although the operative morbidity and mortality is lower for local excision in comparison to pancreatoduodenectomy, there is a greater chance for recurrence of malignant and premalignant neoplasms when the tumor is locally excised. PMID- 3049058 TI - Papillotomy and functional disorders of the sphincter of Oddi. AB - The term "functional" as applied to disorders of the sphincter of Oddi is indefinite since it encompasses a spectrum of disorders from histopathologic fibrosis of the sphincter to sphincteric dysfunction without evident pathologic transformation. Although there are several approaches to the diagnosis of sphincter of Oddi dysfunction, the diagnosis still depends on the exclusion of other causes of obstruction. Reports of surgical sphincteroplasty in patients with this diagnosis suggest that the outcome is unpredictable. Operative mortality ranges from 0.6% to 6.0% and morbidity from 3.3% to 18%. Operative sphincteroplasty appears to have no adverse sequelae long-term, although there are few reports of extended follow-up. In our experience, about 10% of patients undergoing endoscopic sphincterotomy for calculi have recurrent biliary problems during extended follow-up, but most problems were not formidable. In large series of patients undergoing endoscopic sphincterotomy, papillary stenosis is the indication for the procedure in about 5% of cases. With this indication, immediate complications occur with greater frequency compared to endoscopic sphincterotomy for choledocholithiasis. The validity of every proposed test for sphincter dysfunction and/or stenosis has been challenged, and the diagnosis must still be considered imprecise. This fundamental problem makes it difficult to interpret the results of endoscopic sphincterotomy. It also necessitates a cautious approach to management of this disorder by endoscopic sphincterotomy. Other methods of sphincter manipulation such as endoscopic balloon dilation have not been studied extensively. Although endoscopic sphincterotomy has been performed in a few patients with idiopathic recurrent pancreatitis, the procedure must be considered experimental in this group of patients. PMID- 3049060 TI - The use of endoscopic ultrasonography in the diagnosis and staging of carcinoma of the papilla of Vater. AB - Endoscopic ultrasonography (EUS) was evaluated in the diagnosis of carcinoma of the papilla of Vater. Thirteen cases of carcinoma of the papilla examined by EUS were investigated with the aim of assessing the detection capability of EUS in comparison with other diagnostic tools, and its usefulness in diagnosing the extent of the tumor. The EUS image of the normal papilla of Vater presents as a round protruded region with the layered structure of the duodenal papilla along with images of periampullary organs. All but one of 13 tumors of the duodenal papilla presented as a hypoechoic solid mass at EUS. On the basis of an analysis of the layered structure of the papilla of Vater and/or the duodenal wall, EUS images of 12 cases of ampullary carcinoma confirmed histologically were analysed. The EUS images of the tumors of the duodenal papilla corresponded well with the histological findings. When the extent of carcinoma was classified in 4 stages by EUS, the accuracy rate was 83%. EUS is one of the most promising procedures for determining the extent of tumors of the duodenal papilla. PMID- 3049061 TI - Thyroid hormone and the gut. AB - The gastrointestinal tract interacts actively with the thyroid hormones, T4 and T3. Both T4 and T3 are absorbed well but incompletely from the gut, and many factors affect this absorption. The mechanism of absorption is unknown. It is decreased in most malabsorption conditions, but is increased in the postgastrojejunotomy syndrome. It may involve conjugation to the glucuronide forms (T4G and T3G) in the mucosal cell with subsequent deconjugation prior to appearance in the portal vein blood. Absorption appears to be reduced in the presence of excess T4, and increased in hypothyroidism. The liver takes up a large fraction of the T4 and T3 from its circulation and returns a portion of the portal hormone back to the gut via the bile. There is also direct T4 and T3 secretion into the gut from the mesenteric circulation. Recent studies suggest that the gut plays a major role as a reservoir for the thyroid hormones, especially for T3, and that it may also play a role in the regulation of hormone activity. PMID- 3049062 TI - Recent developments in epilepsy. A symposium dedicated to the memory of William G. Lennox, M.D. Seventh annual Merritt-Putnam Symposium. December 7, 1987, Baltimore, Maryland. Proceedings. PMID- 3049063 TI - William G. Lennox: a remembrance. AB - William G. Lennox, author of Epilepsy and Related Disorders, had a lasting effect on our understanding of this illness. He postulated that epilepsy was not a unitary condition and that neuronal chemistries differed from one form of the disease to another. A leader in the use of electroencephalography in epilepsy, he described the first nearly pathognomonic EEG pattern and demonstrated specific features for each of the three most common types of seizure. His pioneering investigations into the biochemical basis of epilepsy helped to identify pathological mechanisms in epileptic attacks. Lennox stood alone in his belief, now generally accepted, that the genetics of epilepsy could be understood only through a multifactorial mode of inheritance. The author presents an affectionate portrait of the physician, the teacher and the man, the founder of the Seizure Unit and the unifying force in the study of epilepsy by both professionals and lay persons. PMID- 3049066 TI - Dysplasminogenemias. AB - This review on dysplasminogenemias describes a growing relationship between genetic polymorphisms of plasminogen and dysplasminogenemias. Plasminogen variants found in eight families in America, Japan and Europe are discussed. Methods used to identify abnormal plasminogens are described in detail. These methods include (a) plasminogen functional to antigen ratios, (b) plasmin generation rates with several plasminogen activators, e.g. urokinase, streptokinase, and the plasmin light (B) chain.streptokinase complex, and (c) plasma and purified plasminogen isoelectric forms. The functional defect including plasminogen kinetics of activation parameters are reviewed, including the formation of plasmin. The molecular defect found in one family, Tochigi I, is described, a single amino acid substitution was found. Finally, the lack of relationships between the abnormal plasminogen variants is reviewed. The variants fall into two classes: one class with a complete absence of a functioning active center, and the second class with different plasminogen kinetics of activation parameters. PMID- 3049065 TI - The relevance of plasminogen activators to neoplastic growth. A review of recent literature. AB - The topics reviewed in this article include transcriptional regulation of plasminogen activators, their relevance to differentiation, tumor progression, and to the metastatic spread of cancer. Recent information on the nature of cellular plasminogen activator receptors is also discussed. Particular attention is paid to the increasing number of observations indicating that several of the inducers of plasminogen activator synthesis utilize distinct regulatory pathways. PMID- 3049064 TI - Effects of in vitro exposure to nitrogen dioxide on human alveolar macrophage release of neutrophil chemotactic factor and interleukin-1. AB - To evaluate the potential toxic and immunologic effects of nitrogen dioxide (NO2) exposure on cells from the lower respiratory tract, normal human alveolar macrophages obtained by bronchoalveolar lavage were exposed to increasing concentrations of NO2 using an in vitro exposure system. Alveolar macrophages exposed to 5, 10, or 15 ppm NO2 for 3 hr showed no difference in cell viability when compared to air-exposed macrophages. In addition, the spontaneous release of neutrophil chemotactic factor (NCF) was not changed by NO2 exposure, nor was there any effect on the ability of alveolar macrophages to release increased amounts of NCF following stimulation with activated zymosan. Furthermore, alveolar macrophages did not spontaneously release interleukin-1 (IL-1) following air or NO2 exposure. When stimulated with influenza virus both air- and NO2 exposed cells released increased amount of IL-1, but was no significant difference in the amount of IL-1 released by air- and NO2-exposed alveolar macrophages. Thus, although NO2 exposure is known to incite an inflammatory response in the lower respiratory tract, using the in vitro exposure system described in this study we were unable to demonstrate a direct toxic effect of NO2 on viability or any NO2-induced change in the release of the immunoregulatory molecules NCF and IL-1. PMID- 3049067 TI - Streptokinase--biochemistry and clinical application. AB - Plasminogen activation to plasmin is due to enzymatic cleavage of a single peptide bond in the zymogen molecule. Streptokinase is not an enzyme and its activation of plasminogen is indirect. Streptokinase forms stoichiometric complexes with plasmin and plasminogen and these complexes activate plasminogen to plasmin. Streptokinase is species selective in its action. PMID- 3049068 TI - The precaution adoption process. AB - This article presents a critique of current models of preventive behavior. It discusses a variety of factors that are usually overlooked-including the appearance of costs and benefits over time, the role of cues to action, the problem of competing life demands, and the ways that actual decision behavior differs from the rational ideal implicit in expectancy-value and utility theories. Such considerations suggest that the adoption of new precautions should be viewed as a dynamic process with many determinants. The framework of a model that is able to accommodate these additional factors is described. This alternative model portrays the precaution adoption process as an orderly sequence of qualitatively different cognitive stages. Data illustrating a few of the suggestions made in the article are presented, and implications for prevention programs are discussed. PMID- 3049070 TI - Immunocytochemical evidence for the cytoplasmic localization and differential expression during the cell cycle of the M1 and M2 subunits of mammalian ribonucleotide reductase. AB - Mammalian ribonucleotide reductase consists of two non-identical subunits, proteins M1 and M2. We have produced and characterized rat polyclonal and monoclonal antibodies directed against protein M2 of mouse ribonucleotide reductase. Using these antibodies for immunocytochemical studies, an exclusively cytoplasmic localization of protein M2 was demonstrated both in cultured parent and hydroxyurea-resistant, M2-over-producing mouse TA3 cells, and in cells from various mouse tissues. These data, together with the previously demonstrated cytoplasmic localization of the M1 subunit, clearly show that ribonucleotide reductase is a cytoplasmic enzyme. Combining the anti-M2 antibodies with a monoclonal anti-M1 antibody allowed for double-labelling immunofluorescence studies of the two subunits in individual cells. Only approximately 50% of the cells in a logarithmically growing culture contained immunodetectable protein M2, while the M1-specific staining was present in all cells. The M2 staining correlates well with the proportion of cells in the S-phase of the cell cycle. In tissues, only actively dividing cells stained with either antibody and there were always fewer cells stained with the M2-antibodies than with the M1-antibody. Our data therefore present independent evidence for the earlier proposed model of a differential regulation during the cell cycle of the M1 and M2 subunits of ribonucleotide reductase. PMID- 3049069 TI - A morphometric study of foetal and newborn cardiac growth in the horse. AB - A morphometric study of hearts in 81 equine foetuses, ranging in age from 190 to 330 days of foetal age, and in 26 newborn foals is reported. The mean weight, external dimensions, ventricular wall thickness and circumference of the atrio ventricular orifice were measured. Features of the main associated vessels of the heart were also recorded. All cardiac measurements of foetuses increased linearly throughout the latter half of pregnancy and were highly correlated with foetal age. This linear growth pattern was also found in the parameters of the associated arterial trunks. The ratio of the right ventricular weight to the total ventricular weight in the equine foetuses increased gradually from 0.27 (Day 190) to 0.34 (full term), but in newborn foals, this ratio began to decrease soon after birth, declining from 0.32 to 0.28 by 11 days of age. The interventricular septum was thickest followed by the right ventricular wall, and the left ventricular wall was thinnest during the period of gestation studied. The right ventricle was 3.2 per cent to 31.5 per cent thicker than the left ventricle. PMID- 3049072 TI - Primary structure and processing of lysosomal alpha-glucosidase; homology with the intestinal sucrase-isomaltase complex. AB - Lysosomal alpha-glucosidase (acid maltase) is essential for degradation of glycogen in lysosomes. Enzyme deficiency results in glycogenosis type II. The amino acid sequence of the entire enzyme was derived from the nucleotide sequence of cloned cDNA. The cDNA comprises 3636 nt, and hybridizes with a messenger RNA of approximately 3.6 kb, which is absent in fibroblasts of two patients with glycogenosis type II. The encoded protein has a molecular mass of 104.645 kd and starts with a signal peptide. Sites of proteolytic processing are established by identification of N-terminal amino acid sequences of the 110-kd precursor, and the 76-kd and 70-kd mature forms of the enzyme encoded by the cDNA. Interestingly, both amino-terminal and carboxy-terminal processing occurs. Sites of sugar-chain attachment are proposed. A remarkable homology is observed between this soluble lysosomal alpha-glucosidase and the membrane-bound intestinal brush border sucrase-isomaltase enzyme complex. It is proposed that these enzymes are derived from the same ancestral gene. Around the putative active site of sucrase and isomaltase, 10 out of 13 amino acids are identical to the corresponding amino acids of lysosomal alpha-glucosidase. This strongly suggests that the aspartic acid residue at this position is essential for catalytic function of lysosomal alpha-glucosidase. PMID- 3049071 TI - Ras-mediated cell cycle arrest is altered by nuclear oncogenes to induce Schwann cell transformation. AB - The cellular responses to ras and nuclear oncogenes were investigated in purified populations of rat Schwann cells. v-Ha-ras and SV40 large T cooperate to transform Schwann cells, inducing growth in soft agar and allowing proliferation in the absence of added mitogens. Expression of large T alone reduces their growth factor requirements but is insufficient to induce full transformation. In contrast, expression of v-Ha-ras leads to proliferation arrest in Schwann cells expressing a temperature-sensitive mutant of large T at the restrictive temperature. Cells arrest in either the G1 or G2/M phases of the cell cycle, and can re-enter cell division at the permissive temperature even after prolonged periods at the restrictive conditions. Oncogenic ras proteins also inhibit DNA synthesis when microinjected into Schwann cells. Adenovirus E1a and c-myc oncogenes behave similarly to SV40 large T. They cooperate with Ha-ras oncogenes to transform Schwann cells, and prevent ras-induced growth arrest. Thus nuclear oncogenes fundamentally alter the response of Schwann cells to a ras oncogene from cell cycle arrest to transformation. PMID- 3049074 TI - Sorting of soluble ER proteins in yeast. AB - In animal cells, luminal endoplasmic reticulum (ER) proteins are prevented from being secreted by a sorting system that recognizes the C-terminal sequence KDEL. We show that yeast has a similar sorting system, but it recognizes HDEL, rather than KDEL: derivatives of the enzyme invertase that bear the HDEL signal fail to be secreted. An invertase fusion protein that is retained in the cells is partially modified by outer-chain mannosyl transferases, which reside in the Golgi element. This supports the view, based on studies in animal cells, that ER targeting is achieved by continuous retrieval of proteins from the Golgi. We have used an invertase fusion gene to screen for mutants that are defective in this sorting system. Over 60 mutants were obtained; eight of these are alleles of a single gene, erd1. The mutant strains grow normally at 30 degrees C, but instead of retaining the fusion protein in the cells, they secrete it. PMID- 3049073 TI - Biogenesis of the yeast lysosome (vacuole): biosynthesis and maturation of proteinase yscB. AB - The biosynthesis and processing of the vacuolar (lysosomal) proteinase yscB was followed in vivo and in vitro. In vitro transcription--translation of the cloned proteinase yscB gene results in a high mol. wt precursor protein (HMr-precursor) of Mr = 73,000. In vivo a precursor of identical mol. wt is found in sec6l mutant cells deficient in translocation of secretory protein precursors into the lumen of the endoplasmic reticulum (ER). In contrast to N-glycosylated intermediate mol. wt forms of proteinase yscB, the HMr-precursor of the enzyme is not N glycosylated, indicating its appearance outside the ER. sec18 Mutant cells, wild type for translocation of proteins into the ER but blocked in the delivery step of secretory proteins to the Golgi apparatus, accumulate a lower mol. wt precursor (LMr-precursor) of proteinase yscB of Mr = 41,500. Processing of the HMr-precursor of proteinase yscB to the LMr-precursor requires proteolytic cleavage which is independent of the proteinase yscA gene product, a protein known to be involved in the processing event of the LMr-precursor of proteinase yscB to the mature enzyme of Mr = 33,000. A LMr-precursor of proteinase yscB of Mr = 42,000 accumulates in proteinase yscA-deficient mutant cells (allele pep4 3). This form is enzymatically active. Incubation in vitro of the LMr-precursor of proteinase yscB with mature proteinase yscA leads to further activation and to processing of the enzyme yielding the mature 33,000 Mr protein.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3049075 TI - Partial purification and substrate analysis of bacterially expressed HIV protease by means of monoclonal antibody. AB - Retroviruses code for a specific protease which is essential for polyprotein precursor processing and viral infectivity. The HIV-specific protease has been predicted to be an aspartic protease which is located at the amino terminus of the pol gene. We have prepared several constructs for bacterial expression of the protease. Two of them span the whole protease region and result in its autocatalytic activation. Analysis of the dynamics of this activation indicates a two-step process which starts at the carboxy terminus and ends at the amino terminus of the protease. The activated protease is a molecule of 9 kd as evidenced by monoclonal antibody in immunoblot analysis. A construct in which the carboxy terminus of the protease is deleted results in a stable, enzymatically inactive 27-kd protein which proved useful as substrate since it contains one of the predicted cleavage sites. The stability of this protein indicates that the carboxy-terminal sequences of the protease are essential for its activity and its autocatalytic activation. The protease which is very hydrophobic was solubilized by acetone treatment and passaged over ultrogel and propylagarose columns for partial purification. It elutes as a dimer and tends to aggregate. It is inhibited by pepstatin A in agreement with its expected active site and its theoretical classification as aspartic protease. Cleavage of the gag precursor results in the mature capsid protein, p17. The protease does not, however, cleave the denatured 27-kd substrate or the denatured gag precursor. Therefore its specificity appears to be not solely sequence- but also conformation-dependent. This property needs to be taken into account for the development of protease inhibitors for therapy of AIDS. PMID- 3049076 TI - Multiple regulatory mechanisms control the expression of the RAS1 and RAS2 genes of Saccharomyces cerevisiae. AB - Expression of the RAS1 and RAS2 genes of Saccharomyces cerevisiae has been examined at the transcriptional and translational levels. When dextrose is the carbon source, the steady-state amount of RAS1 mRNA and the rate of RAS1 protein synthesis are reduced in parallel as cells approach the mid-exponential phase of growth. RAS1 mRNA levels and protein synthesis are very low at all stages of growth when ethanol rather than dextrose is provided as the sole carbon source. The rate of RAS2 protein synthesis is regulated differently. In cells cultured on dextrose, it is lowest in the early exponential phase, increases approximately 10 fold and remains nearly constant as cells approach stationary phase. By contrast, RAS2 mRNA is found at uniformly high levels at all phases of exponential growth, suggesting that the translational efficiency of RAS2 mRNA is repressed during the early exponential phase. This repression is not observed when ethanol is the sole carbon source. Nutrient starvation, resulting in G1 arrest and sporulation in diploids, leads to greatly decreased amounts of RAS2 mRNA, accomplished in part by selective repression of RAS2 transcripts with particular 5' ends. However, this reduction in RAS2 mRNA levels has little effect on the rate of RAS2 protein synthesis, suggesting that the translational efficiency of RAS2 mRNA is stimulated by nutrient starvation. The combination of transcriptional and translational controls which regulate yeast RAS gene expression seems to ensure that one or the other RAS proteins will be produced over a wide range of physiological states. PMID- 3049077 TI - ProOmpA spontaneously folds in a membrane assembly competent state which trigger factor stabilizes. AB - The precursor protein proOmpA can translocate across purified Escherichia coli inner membrane vesicles in the absence of any other soluble proteins. ProOmpA, purified 2000-fold in the presence of 8 M urea, is competent for translocation following rapid renaturation via dilution. ATP, the transmembrane electrochemical potential, and functional secY protein are essential for the translocation of proOmpA renatured by dilution. The kinetics of its translocation and the level of translocation at each concentration of ATP are indistinguishable from that of proOmpA renatured by dialysis with trigger factor. After dilution, the proOmpA rapidly loses its competence for membrane assembly. However, this competence is stabilized by trigger factor. Assembly-competent proOmpA is in a protease sensitive conformation, whereas proOmpA which has lost this competence is more resistant to degradation. This suggests that the primary role for trigger factor in in vitro protein translocation is to maintain precursor proteins in a translocation-competent conformation. We propose that a properly folded precursor protein and ATP are the only soluble components which are essential for bacterial protein translocation. PMID- 3049078 TI - The export of the DNA replication inhibitor Microcin B17 provides immunity for the host cell. AB - Microcin B17 (MccB17) is a peptide antibiotic which inhibits DNA replication in Enterobacteriaceae. Microcin-producing strains are immune to the action of the microcin. Physical and genetic studies showed that immunity is mediated by three genes: mcbE, mcbF and mcbG. We sequenced these genes and identified polypeptide products for mcbF and mcbG. By studying the contribution of each gene to the expression of immunity we found that immunity is determined by two different mechanisms. One of these, encoded by mcbE and mcbF, is also involved in the production of extracellular MccB17. To reconcile these observations we propose that McbE and McbF serve as a 'pump' for the export of active MccB17 from the cytoplasm. This model is supported by the predicted properties of the McbE and McbF proteins, which are thought to be, respectively, an integral membrane protein and an ATP-binding protein with homology to other transport proteins. PMID- 3049079 TI - Correlation between the conformation of Escherichia coli -10 hexamer sequences and promoter strength: use of orthophenanthroline cuprous complex as a structural index. AB - The lac and gal control regions contain two functional overlapping promoters P1 and P2. Point mutations can shift transcription from P1 to P2 and vice versa. We show that the reactivity of DNA fragments towards nucleolytic attack with orthophenanthroline cuprous complex can be used to predict which promoter competes more efficiently for RNA polymerase binding. Furthermore, similar changes in reactivity are observed as closed complexes isomerize to form the final open complexes, provided that the functional start is taken as a reference. We found a correlation between the reactivity pattern of -10 regions in uncomplexed DNA and the rate of open complex formation. PMID- 3049081 TI - Cooked-Food Mutagen Reference List and Index. PMID- 3049082 TI - Enhancer sequences and the regulation of gene transcription. PMID- 3049080 TI - Recognition of the P1 plasmid centromere analog involves binding of the ParB protein and is modified by a specific host factor. AB - The P1 plasmid partition system is responsible for segregation of daughter plasmids during division of the Escherichia coli host cell. The P1-encoded elements consist of two essential proteins, ParA and ParB, and the cis-acting incB region. The incB region determines partition-mediated incompatibility and contains the centromere-like site parS. We have isolated and purified the two proteins. ParB binds specifically to the incB region in vitro. DNase I footprinting assays place a strong binding site over the 35-bp parS sequence previously shown to be sufficient for partition when the Par proteins are supplied in trans. A weaker site lies within the incB region in sequences that are important for specifying incompatibility, but are not essential for partition. Gel band retardation assays show that a host factor binds specifically to the incB sequence. The factor strongly stimulates binding of ParB. Cutting the region at a site between the two ParB binding sites yields two fragments that can bind ParB but not host factor. Thus, information for host-factor binding lies in the region determining the specificity of plasmid incompatibility. The roles of parB and the host factor in partition and the specificity of plasmid incompatibility are discussed. PMID- 3049083 TI - Citric-acid cycle, 50 years on. Modifications and an alternative pathway in anaerobic bacteria. AB - Many anaerobic bacteria can completely oxidize organic matter to CO2 with either sulfur, sulfate, or protons as electron acceptor. The sulfur-reducing bacteria and one genus of sulfate reducers use a modified citric-acid cycle with a novel anaplerotic sequence as pathway of terminal respiration. All other anaerobes use an alternative pathway, in which carbon monoxide dehydrogenase is a key enzyme and in which acetyl-CoA is cleaved into two C1 units at the oxidation level of CH3OH and CO. Thus almost 50 years after the discovery of the citric acid cycle by Hans Krebs in 1937, a second pathway for acetyl-CoA oxidation was found. PMID- 3049084 TI - Intravenous digital subtraction angiography of the precerebral and cerebral arteries in patients with TIA. A comparison with conventional angiography. AB - A study of the diagnostic utility of both intravenous digital subtraction angiography (IV-DSA) and conventional angiography of precerebral and cerebral arteries is presented. The series comprised 60 patients with TIA, who underwent the two procedures with a mean interval of 16 days. Conventional angiography was generally superior to IV-DSA, and this was particularly marked in the siphons and cerebral arteries. Only the excellent IV-DSA examinations obtained in a few patients with TIA could be accepted as final pre-operative procedures. Accurate imaging of the lesions and collateral flow pattern usually required intra arterial injections, and intra-arterial DSA is now usually preferred. PMID- 3049085 TI - Spontaneous arterio-venous fistulae of the vertebral artery. Diagnostic and therapeutic considerations. AB - Spontaneous arterio-venous fistulae of the vertebral artery are rare. These lesions mainly affect the upper cervical area, and are usually asymptomatic, or may present as small, often pulsatile, cervical masses with vascular murmurs. The authors report on two cases in which the presumptive diagnosis, suggested by venous digital subtraction angiography, was then confirmed by selective angiography. In both cases an intravascular approach with detachable balloons and particulate substances was carried out, with good anatomical and functional results. Problems related to diagnosis, pathophysiology of symptoms, indications for treatment and embolization techniques are discussed. PMID- 3049086 TI - Infundibulopelvic stenosis--evaluation of diagnostic imaging. AB - Infundibulopelvic stenosis is a very rare kidney disease. This congenital disorder must be differentiated from multicystic dysplastic kidney, polycystic kidney disease, simple renal cysts and mega-(poly)-calicosis. Associated abnormalities of the ureter are rare. Ureteric obstruction was associated in our two patients, and in one case agenesis of the bladder and a duplicated genital tract were also present. PMID- 3049088 TI - Ultrasound in the detection of adrenal tumours. AB - An analysis was made of the ultrasound (US) findings in 110 patients with proven or ultrasonographically suspected adrenal tumours. A total of 212 glands were imaged, 117 of which contained a tumour. The diagnosis was confirmed histologically in 50 cases and by imaging and follow-up in 162 cases. Normal glands were visualized ultrasonically in 27 out of 40 glands on the right and 5 out of 55 on the left. US detected 55 out of 59 primary adrenal tumours and 49 out of 58 secondary tumours, showing a sensitivity of 89%. In 13 tumours ultrasonography gave a false negative result, and in 13 cases a false positive one. Benign or primary malignant tumours and metastases could not be differentiated on the basis of echo-structure, but a large heterogeneous tumour in a patient with no known extra-adrenal malignancy is probably a primary malignant adrenal tumour. PMID- 3049087 TI - Digital subtraction radiography in voiding cystourethrography. AB - Digital subtraction radiography (DRS) was utilized to evaluate the anatomy and function of the bladder and urethra. Images were obtained in 30 patients, following urography or retrograde cystography, with full bladder distention and during different phases of micturition. The technique permits good demonstration of several anatomical and functional parameters (lowering of bladder base; dynamic origin of trigonal canal; rotation of the urethra; bladder wall motion). PMID- 3049089 TI - Altered growth factor requirements and cell cycle control in rat hepatoma cells versus adult rat hepatocytes in culture. AB - Adult rat hepatocytes multiply in primary cultures when incubated in arginine free MX-83 medium supplemented with dialyzed fetal calf serum, insulin, glucagon, hydrocortisone, epidermal growth factor, and transferrin. In the absence of mitogens, the fraction of the cells engaged in DNA synthesis dropped sharply. However, cells initiated DNA synthesis in response to the mitogenic mixture indicating that hepatocyte proliferation is controlled by G1----S transition rates. In contrast, rat hepatoma line DTH-3, derived from Morris 7777 "minimal deviation" hepatoma, required only insulin for proliferation in chemically defined MX-83 medium. The lengths of their cell cycle phases varied with the growth rate. The phases of the growth cycle were proportionately shortened (expanded) when the growth rate was increased (decreased). It is concluded that DTH-3 hepatoma cells, which display a decreased growth factor requirement as compared with adult rat hepatocytes differ from normal hepatocytes by fundamental alterations in the mechanisms controlling the progression of the cell cycle. PMID- 3049090 TI - Angiotensin converting enzyme inhibitors and ischaemic heart disease. PMID- 3049091 TI - Value of radionuclide assessment with thallium 201 scintigraphy in carnitine deficiency cardiomyopathy. AB - The case of a 30 month-old boy who presented with isolated severe dilated cardiomyopathy is reported. The diagnosis of systemic carnitine deficiency was confirmed by low serum and tissue carnitine levels. During oral L-carnitine therapy, dramatic improvement of the cardiac function was assessed by radionuclide methods. Myocardial thallium 201 uptake was closely correlated with cardiac function studied by angioscintigraphy. These methods are simple, easily reproducible, non-invasive and involve little radiation. In a case of cardiomyopathy, we suggest an immediate trial of oral carnitine treatment; the efficacy of the therapy can be confirmed by isotopic tests with thallium 201 scintigraphy. PMID- 3049092 TI - Intracellular mechanism of action of vasodilators. AB - Vasodilators that act directly at the level of the vasculature may be classified as either endothelium-dependent or endothelium-independent agents. Endothelium dependent agents stimulate the endothelium to produce endothelium-derived relaxing factor (recently identified as NO) which relaxes vascular smooth muscle and increases cGMP. Endothelium-independent vasodilators may be divided into calcium antagonists, alpha 1-adrenergic antagonists, cAMP-elevating agents and cGMP-elevating agents. It is generally accepted that calcium-dependent phosphorylation of myosin light chains initiates smooth muscle contraction. A variety of evidence suggests that certain vasodilators inhibit calcium-dependent phosphorylation of myosin light chains and smooth muscle contraction via activation of cAMP- and cGMP-dependent protein phosphorylation. Since vascular smooth muscle cells and platelets have many properties in common, and since cAMP- and cGMP-elevating vasodilators inhibit both smooth muscle contraction and platelet aggregation, human platelets were used to study the effects of vasodilators on protein phosphorylation. It could be demonstrated that cAMP- and cGMP-regulated protein kinases are the intracellular mediators for prostaglandin E1- and nitroprusside-induced protein phosphorylation, respectively. Furthermore, prostaglandin-E1- and nitroprusside-stimulated protein phosphorylation was found to be associated with an inhibition of the phosphatidylinositol cycle. Cyclic nucleotide-elevating vasodilators may inhibit smooth muscle contraction and platelet aggregation by regulating the flux through the phosphatidylinositol cycle, and the activity of myosin light-chain kinase and of calcium ATPases. PMID- 3049093 TI - Calcium antagonists in heart failure. AB - Calcium channel blocking drugs improve systolic function, due to a decrease in left ventricular afterload, and may thus be particularly suitable for therapy of patients with concomitant congestive heart failure and myocardial ischaemia. Since ischaemic heart disease is a major cause of congestive heart failure and further ischaemia may contribute to further left ventricular dysfunction, studies were performed to establish the therapeutic role of calcium channel blockers. Concern has been raised that at high doses, calcium channel blockers have a negative inotropic effect and may lead to deterioration of myocardial systolic function. Clinical studies assessing the role of verapamil and diltiazem in congestive heart failure are scarce, because these drugs may result in a further decrease in cardiac output, especially in the presence of pre-existing left ventricular dysfunction. In vitro observations show that nifedipine is more potent on vascular smooth muscle than on myocardium. It has been suggested that in man, nifedipine does not usually reach concentrations high enough to depress myocardium and the vasodilating left ventricular unloading effect becomes dominant. Analysis of the available clinical data is difficult, but published data concerning the oral or sublingual administration of nifedipine in congestive heart failure suggest that the major haemodynamic effect is a decrease in systemic vascular resistance by 33%, resulting in an increase in cardiac output of 24%. Heart rate was generally unchanged. Right atrial pressures showed minimal change, whereas pulmonary capillary wedge pressure decreased by 16%. The response is more favourable if the congestive heart failure is more severe. A new dihydropyridine, nisoldipine, was evaluated during chronic administration over 8 weeks in patients with severe congestive heart failure.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3049094 TI - Vascular and renal prostaglandins as counter-regulatory systems in heart failure. AB - Vasoconstrictors and vasodilators are both activated in patients with severe heart failure. Vasodilatory prostaglandins are increased in parallel with the degree of activation of neurohumoral vasoconstrictor systems, and may serve to offset the circulatory effects of systemic and regional vasoconstriction. Enhanced vasoconstrictive and vasodilatory activities appear to be especially important in patients with hyponatraemia. These patients may be particularly susceptible to clinical deterioration when given drugs that inhibit prostaglandin synthesis (e.g. indomethacin) and may be more likely to improve when treated with agents that enhance the production of endogenous prostaglandin (e.g. angiotensin converting enzyme inhibitor). Exogenous PGE2 reduces afterload and may be a useful therapeutic agent. Research on vascular and renal prostaglandins should further our knowledge of the pathophysiology and therapy of congestive heart failure. PMID- 3049095 TI - The origin of symptoms in patients with chronic heart failure. AB - Severe shortness of breath is a prominent symptom in acute heart failure (pulmonary oedema) and is related to left atrial pressure. A reduction of this pressure almost always leads to an improvement in symptoms. Patients with chronic heart failure complain of both shortness of breath and tiredness even when fluid overload has been corrected by the appropriate use of diuretics. Shortness of breath under these circumstances is not related simply to central haemodynamics but is determined more by the interaction of changes in respiratory pattern and the metabolic consequences of reduced perfusion of exercising skeletal muscle. An important clinical consequence is that when such patients are optimally treated with diuretics, further improvement of symptoms would not be expected from drugs which merely alter central haemodynamics without influencing other factors such as skeletal muscle blood flow on exercise, or lung perfusion. PMID- 3049096 TI - Role of vasopressin in experimental heart failure. AB - Neurohumoral vasoconstrictor systems are activated in heart failure and influence left ventricular function by modifying pre- and afterload. The renin-angiotensin system, sympathetic nerve activity and vasopressin have all been implicated as mechanisms of vasoconstriction. We investigated the vasoconstrictive action of vasopressin by blocking its vascular receptors by a specific antagonist in animal models of heart failure due to rapid right ventricular pacing in dogs and to aortocaval fistula, pulmonary stenosis and aortic stenosis in rats. In all animal models studied, we observed an inappropirately elevated secretion of vasopressin in association with a decreased plasma osmolality. We found only a small reduction of aortic pressure and peripheral vascular resistance following the vasopressin inhibitor. The renin-angiotensin system and sympathetic nerve activity were activated in all animal models. We injected teprotide in order to test the role of angiotensin in the regulation of peripheral vascular tone, and found a much greater decrease of blood pressure and peripheral vascular resistance than caused by the vasopressin antagonist. These experiments suggest that vasopressin and the renin system increase peripheral vascular tone in certain models of heart failure, and demonstrate a much greater importance for the renin system. PMID- 3049097 TI - Converting enzyme inhibitors in heart failure. AB - The renin-angiotensin system is activated in heart failure in proportion to the severity of the haemodynamic derangement and to diuretic dose. Angiotensin converting enzyme (ACE) inhibitors reduce circulating levels of angiotensin II and aldosterone and, in some patients, plasma noradrenaline, vasopressin and cortisol. Typically there is potassium retention and a minor increase in plasma potassium, but cumulative sodium balance may increase or decrease depending on pretreatment fluid and haemodynamic status and on policy regarding diuretic dose. Circulatory dynamics usually improve and blood flow to the brain, myocardium and kidneys is preserved. Changes in glomerular filtration rate are dictated by haemodynamic characteristics and, again, by diuretic dose and dietary sodium. There are potential hazards with ACE inhibitor therapy but most problems can be anticipated and avoided. Future trends may include the introduction of ACE inhibitors with or without concomitant diuretic therapy in early cardiac failure, and intravenous ACE inhibition immediately after acute myocardial infartion. Whether the ACE inhibitors will prove more successful than alternative antihypertensive agents in preventing cardiac complications (including heart failure) of hypertension, is an intriguing question. PMID- 3049098 TI - Non-receptor-mediated inotropic drugs. AB - There has been an active search recently for non-glycoside, non-sympathomimetic positive inotropic agents. Phosphodiesterase inhibitors inhibit the breakdown of cyclic AMP, leading to an increase in intracellular cAMP concentration, and can be expected to enhance the force of myocardial contraction. The methylxanthines exert such an action in vitro; the situation in vivo is more complex, due to their manyfold actions. Phosphodiesterase F-III is relatively specific for cAMP degradation; the new phosphodiesterase inhibitors may act specifically by inhibiting this enzyme. Phosphodiesterase inhibitors with combined inotropic and vasodilatory action include amrinone, which is no longer in widespread use due to its pronounced side-effects, milrinone, which is much better tolerated and has shown promising results in recent large-scale trials, and sulmazole which has been withdrawn due to toxic effects in rodents. Other drugs are still under investigation. Of importance is the relative role of the inotropic versus dilatory action of these drugs. A salutory effect on resting haemodynamics, does not necessarily imply improved exercise haemodynamics. The pharmacokinetic profile of these drugs is also of interest. The clinical benefits should be sustained and the side-effects should not outweigh the beneficial actions of these drugs. PMID- 3049099 TI - Mechanism of action of calcium antagonists in heart and vascular smooth muscle. AB - A transmembrane supply of Ca2+ ions is required for active tension development in both vascular smooth musculature and myocardial fibres. Ca2+ antagonists thus not only damp hyperkinetic myocardial dysfunction but also act against practically all vasoconstrictor or spastic responses of extramural coronary smooth muscle. Clinically important targets of Ca2+ antagonists include the pulmonary, cerebral, mesenteric and renal arteries, as well as the peripheral resistance vessels of the systemic circulation. Ca2+ antagonists are used increasingly to treat acute hypertensive crises as well as for long-term antihypertensive therapy. In physiological experiments, Ca2+ antagonists not only neutralize various vasoconstrictor agents but also greatly reduce the sensitivity of the systemic arteries and arterioles to mechanical stimuli such as extension of the vascular wall by a rise in intraluminal pressure. In human arterial walls, at an advanced age, cytotoxic degrees of Ca2+ overload probably play an important role in the pathogenesis of arteriosclerotic lesions. Severe diabetics and heavy smokers exhibit an even faster progression of age-dependent arterial calcinosis. In rats, the Ca2+ antagonist verapamil not only prevented arterial calcinosis due to overdoses of vitamin D and dihydrotachysterol but also counteracted age-dependent Ca2+ accumulation. Spontaneously hypertensive rats also exhibit progressive arterial Ca2+ overload which responds excellently to the Ca2+ antagonists nifedipine, nimodipine, nisoldipine, nitrendipine, as well as to verapamil and diltiazem. Suitable Ca2+ antagonists could possibly retard or even prevent premature arterial calcinosis in humans. Moreover, Ca2+ antagonists exert a direct cardioprotective effect on the myocardium by preventing intracellular Ca2+ overload. PMID- 3049100 TI - Non Hodgkins lymphoma of the breast, unusual presentation: detection by 67Ga scintigraphy. AB - Lymphomatous involvement of the breast is an uncommon cause of breast masses. A case is presented of a patient with bilateral breast involvement revealed by intense 67Ga uptake. Other foci of involvement were also detected scintigraphically, and confirmed by other imaging modalities. Multiagent chemotherapy resulted in significant clinical and scintigraphic regression of tumor, demonstrating the potential utility of 67Ga imaging in the follow-up of these patients. PMID- 3049101 TI - Terguride in parkinsonism. A multicenter trial. AB - Terguride is an ergoline derivative with mixed agonistic/antagonistic dopaminergic activity. This led to a paradoxical suggestion that it is effective in the treatment of both schizophrenia and parkinsonism. A total of 65 in- or outpatients with parkinsonism mostly of vascular or idiopathic etiology were included in a 4-week, open, multicenter trial. Terguride was administered under an increasing dose schedule which was leveled off according to the clinical response. Mostly because of nausea, vomiting, and lack of improvement 25% of inpatients and 61% of outpatients were removed from the study. The average daily dose at the end of the trial was 4.2 mg, ranging from 1.0 to 5.5 mg. The average Simpson and Angus scale total score and performance in the Spiral Drawing Task improved significantly during the trial by 20% and 38% respectively. The following adverse effects were noted most frequently throughout the study (including those who withdrew): constipation (occurred in 42% of all ratings performed during the trial) drowsiness and nausea (16% each). Adverse circulatory effects were negligible. Psychotic symptoms, including depression, confusion, hallucinations, and paranoid syndrome, each occurred in 1 patient, i.e., at a lower rate than with other dopaminergic drugs. Scotopic electroretinograms in a subsample of 7 patients showed a significant transitory decrease in the B-wave amplitude at the end of the 1st week and a subsequent return to pretreatment values. PMID- 3049102 TI - Intrarectal bypass graft in low anterior resection and sigmoid obstruction--an experimental study. AB - The most common causes for morbidity and mortality in colorectal resections are anastomotic leaks. In low anterior resection, the incidence of anastomotic leakage ranges from 17 to 50%. With the use of the stapler technique, leakage incidence rate remains high and ranges from 10 to 25%. Colostomy formation and closures are associated with considerable morbidity and mortality. Due to the high incidence of anastomotic leakage rate in low anterior resection, and the additional complications of diverting colostomy formation and closure, the use of a rectal stent-intrarectal bypass graft has been instituted. This is carried out by means of a silastic graft, which prevents the fecal stream and gas pressure from coming into contact with the anastomotic site at the low rectum. The efficacy of intrarectal bypass graft was examined in two high-risk surgical situations, the first in very low anterior resection and the other, after early sigmoid obstruction. In both situations the intrarectal bypass graft provided for a safe anastomosis. Even when dehiscence and early obstructions occur, the tube may prevent leakage. This procedure presents effective practical implications which obviate the need for a proximal colostomy formation, thereby eliminating the physical and psychological stress that accompanies colostomies. PMID- 3049104 TI - Local shock-wave lithotripsy of distal ureteral calculi. AB - Since the initiation of the clinical trial utilizing a second-generation lithotripor (Lithostar, Siemens, Erlangen, FRG), 96 patients with distal ureteral calculi (i.e. calculi below the pelvic brim) underwent local shock-wave lithotripsy. Routine treatment was conducted under intravenous sedation and light analgesia only. Complete stone disintegration was achieved in 84 patients (87.5%), 11 requiring two sessions and 1 patient, three. In 7 patients ureteroscopy became necessary after unsuccessful local shock-wave treatment. In 2 of these patients a 9-french flexible ureteroscope and the Storz Q-switched neodymium-YAG laser was used for stone disintegration. In 3 cases loop extraction and in 2 cases open surgery had to be performed for definitive stone removal. All pre- and postoperative manipulations (except open surgery) were done on the Lithostar. Local shock-wave lithotripsy is a highly successful, noninvasive, time saving and easily applicable technique. It has become our primary approach in the treatment of distal ureteral calculi. PMID- 3049103 TI - Effect of ciclosporin on allogeneic hepatocyte transplantation: a morphological study. AB - Hepatocyte transplantation (HT) is among the most frequently employed approaches in the treatment of liver disease. This work deals with the occurrence of rejection when allogeneic hepatocytes are transplanted into the spleens of rats. Two strains of rats were used: Lewis and F344. The recipients were divided into two groups, one of which received ciclosporin (Cic), while the other received no treatment. After HT, the spleens were removed periodically. The animals that did not receive Cic suffered such intense rejection that 7 days after HT no hepatocytes could be identified in the splenic pulp. On the other hand, hepatocytes could be observed throughout the entire study in the spleens of rats treated with Cic. We conclude that HT is followed by intense rejection, which can be avoided by the administration of Cic. PMID- 3049105 TI - Impact of sonography on diagnosis of scrotal diseases: a multicenter study. AB - Scrotal ultrasound was carried out since 1980 in five centers. 1,971 cases presenting with pathological conditions of scrotal content were reviewed to asses limitations and possibilities of the method. Scrotal sonography was found to be a highly reliable method to distinguish between intra- and extratesticular lesions. The data presented reveal that sonography is not capable of differentiating reliably between benign and malignant lesions. In the decision-making process ultrasound is sometimes of great help to the urologist but surely no substitution for careful examination and history taking. PMID- 3049107 TI - IgG and IgA antibodies specific for Chlamydia trachomatis in acute epididymitis. AB - The possibility of using elevated Chlamydia trachomatis-specific serum IgG and IgA as a screening test for Chlamydia-associated epididymitis was analyzed in 28 acute epididymitis patients and 42 apparently healthy men by the single antigen (L2) immunoperoxidase assay. The prevalence rates of C. trachomatis IgG antibody titer greater than or equal to 64 and elevated C. trachomatis IgG titers (greater than or equal to 128) were significantly higher in the epididymitis patients (75 vs. 40%, p less than 0.01, and 39 vs. 14%, p less than 0.025, respectively) than in controls. The prevalence rate of C. trachomatis IgA antibodies (titer greater than or equal to 8) was significantly higher (p less than 0.001) in epididymitis patients as compared to controls (46 vs. 10%, respectively). The potential application of elevated serum C. trachomatis IgG and IgA antibodies as a noninvasive screening marker in epididymitis patients is discussed. PMID- 3049106 TI - A progressive radiorenographic (99mTc-DTPA) presentation of surgically induced renal vein hypertension in baboons. AB - A 99mTc-DTPA radiorenographic study, supported by excretory urography, blood chemistry and eventually histology was performed on 8 chacma baboons (Papio ursinus) after surgical induction of unilateral renal vein hypertension. Frequent sequential repeat renograms during 6 post-operative months detected fluctuating renal behaviour but did not reveal typical course of the disease. Neither did any one of the radiorenographic parameters (TP, T1/2, FF, relative clearances and perfusion) prove to be more sensitive than the others in detecting abnormalities. Initial microscopic or macroscopic haematuria and proteinuria in general eventually cleared up. Light microscopy demonstrated only mild abnormalities. The effects of venous occlusion of the kidney and the prognosis seemed to be variable as was reflected by radiorenography. PMID- 3049109 TI - Digital examination and transrectal ultrasonography in the diagnosis of prostatic cancer. AB - The efficiency of digital examination and transrectal ultrasonography in the diagnosis of prostatic cancer was examined in 109 untreated male patients with symptoms of urinary obstruction, including 32 cases of prostatic cancer. The sensitivity and specificity of digital examination were 69 and 79%, respectively, with an efficiency of 76%; while, the sensitivity and specificity of ultrasonography were 88 and 77%, respectively, with an efficiency of 80%. Of the 10 cases of subclinical cancer, where the patient had a clinically benign gland with histological evidence of cancer, 8 were correctly diagnosed ultrasonographically. Five of the 8 cases could be diagnosed by the presence of a focal hypoechoic lesion in the unilateral lobe of the prostate. These results indicate that transrectal ultrasonography is more efficient than digital examination for the diagnosis of prostatic cancer and that hypoechoic change is an important point for detecting such subclinical cancer on ultrasonography. PMID- 3049108 TI - Percutaneous management of urolithiasis after kidney transplantation. Report of a case and review of the literature. AB - Urolithiasis is one of the least common urological complications after kidney transplantation, but it remains an important cause of deterioration of graft function. We report a case managed by percutaneous nephrostolithotomy and review the literature. PMID- 3049110 TI - Sequential study of macrophage migration inhibition factor in renal cell carcinoma patients. AB - Preliminary investigation of cell-mediated immunity (CMI) in 49 patients harboring renal cell carcinoma (RCC) by the migration inhibition factor (MIF) tests as compared to 47 patients with simple renal cysts or normal kidneys established the specificity (85%) and reproducibility of this test. A retrospective analysis of the results of the MIF test was conducted in 35 patients followed for a minimum 20-month period. A marked difference was observed between the group of patients with localized disease who remained disease free and those with metastatic lesions and disease progression. The first group sustained their positive response of the MIF tests for a longer time during the study period, while the latter had a decrease of the MIF test response shortly after surgery. This difference may indicate that immune response as reflected in the positive MIF test has a role in RCC patients' defense mechanism. PMID- 3049111 TI - Reappraisal of antibiotic susceptibility tests in the management of chronic bacterial prostatitis. AB - In view of 50 cases of chronic bacterial prostatitis the efficacy and limitations of antibiotic susceptibility tests have been evaluated and factors affecting passage of drugs across the prostatic cell membrane have been discussed. PMID- 3049112 TI - Interventional radiology in oncology. PMID- 3049113 TI - The role of heterochromatin in the origin of isochromosome 1 in neoplastic cells. PMID- 3049114 TI - Neuron-specific enolase: how useful as a cancer marker? PMID- 3049115 TI - Identification of a malignant cell associated antigen recognized by a human monoclonal antibody. AB - A human X human hybridoma, CLNH11, derived from a lymphocyte of a patient with cervical carcinoma, produces a human monoclonal antibody of gamma 1 and kappa isotypes (CLN-IgG). Immunoperoxidase staining showed that CLN-IgG reacted with frozen tissue sections of human malignant tumors (cervical carcinoma, gall bladder carcinoma, glioblastoma), but not with their normal counterparts. Enzyme linked immunosorbant assay also demonstrated that CLN-IgG reacted with various human tumor cell lines, but not with non-tumorigenic cells such as some fibroblasts, peripheral blood lymphocytes and red blood cells. Indirect and direct immunofluorescence staining indicated that the tumor antigens recognized by CLN-IgG were located in restricted areas close to the cell surface and exposed on the outer surface of the cell membrane. A protein antigen of Mr 226,000 was purified to homogeneity by affinity chromatography with CLN-IgG from the plasma membrane fraction of A549 lung tumor cell line. The antigen consisted of alpha (Mr 60,000) and beta subunit (Mr 53,000) which were linked by disulfide bond(s) (TA60K/53K). The TA60K/53k antigens were expressed commonly in other tumor cell lines originated from histologically different tissues. PMID- 3049116 TI - Transfer factor in Hodgkin's disease: a randomized clinical and immunological study. AB - Transfer factor (TF) was prepared from buffy coats obtained from 493 units of blood taken from healthy donors, including individuals convalescent from various viral infections. It was administered to 22 of 47 patients with Hodgkin's disease undergoing treatment and consenting to take part in this randomized study to determine if TF would enhance their immunity and/or reduce the incidence of subsequent infections. Skin test reactivity was markedly enhanced in those patients receiving TF as opposed to placebo but other immunological assessments showed no significant differences between the groups. TF was not shown to be of benefit in the prevention of infections (including varicella/zoster). PMID- 3049118 TI - The efficacy and tolerability of atenolol, nifedipine, and their combination in the management of hypertension. AB - In this randomized, double-blind, cross-over study we investigated the haemodynamic effects of a beta-blocker (atenolol 50 mg) and a calcium antagonist (nifedipine SR 20 mg) given either separately or in combination in three groups of hypertensive patients. Each treatment was administered twice daily. The fixed combination given twice daily for four weeks produced reductions in blood pressure which lasted for at least 12 h after administration of the last dose. The control of blood pressure by the combination was superior to that achieved by its individual components. Adverse effects normally associated with nifedipine were less frequent when it was given with atenolol. Compliance with treatment was good, but best when the drugs were given together rather than separately. A fixed combination of atenolol and nifedipine may prove useful in treating hypertensive patients inadequately controlled on beta-blocker therapy alone. PMID- 3049117 TI - Dosage regimen of cimetidine reviewed. Possible drug accumulation after multiple oral doses. AB - The doses of cimetidine recommended differ in children, especially those with cystic fibrosis. These dosage regimens were derived from single-dose pharmacokinetic studies of the drug. Some authors showed, however, that after administration of repeated oral doses of cimetidine in healthy adults and children with cystic fibrosis, the elimination half-life of the drug was markedly prolonged. In view of the ability of cimetidine to inhibit metabolism of other drugs, it is suggested that the parent compound and/or it is metabolite(s) may inhibit its own metabolism during a prolonged course of treatment. Enterohepatic recirculation of the drug and/or its metabolite(s) may also contribute to prolongation of its elimination. One should therefore be cautious in using single dose pharmacokinetic parameters to calculate repeated dose regimens and expected plasma steady-state concentrations. PMID- 3049119 TI - B cell-stimulating activity of lymphoid cell membrane fractions. AB - We had previously found that a mutagenized subline of the mouse thymoma EL4 very efficiently stimulates B cells via direct cell-cell contact, thereby inducing the responsiveness of B cells to cytokines. In the present study, we investigated whether this effect could also be mediated by plasma membranes of EL4 (and other) cells. By equilibrium centrifugation of cell homogenates, four cell membrane fractions of different densities were obtained. These were tested for (a) stimulation of B cell proliferation in conjunction with EL4 supernatant as source of cytokines, and (b) enhancement of B cell proliferation at suboptimal concentration of lipopolysaccharide. It turned out that all membrane fractions from a variety of T lineage cells (mutant EL4, parent EL4, BW5147, P198 thymomas, normal T cells) and B lineage cells (BCL1 lymphoma, X63Ag8 cytoplasma, normal B cells) exhibited similar B cell stimulating activity in both assays. Interleukin 1 activity was not detected in the membrane fractions. Heat treatment abolished all activity showing that protein at least was involved. Either protease treatment or extraction with detergent abolished the activity of subcellular fractions rich in intracellular membranes but not that of fractions most enriched in surface membranes. Finally, erythrocyte membranes also displayed B cell stimulating activity sensitive to protease and detergent extraction. In contrast, and in confirmation of a previous study, liver cell membrane was inhibitory in the B cell proliferation assay with lipopolysaccharide. In conclusion, the effects of cell membranes did not reflect the unique activity of intact mutant EL4 cells. However, with respect to our data it is conceivable that membrane proteins with relatively nonspecific activity and wide distribution among lymphoid cells could play a role in T cell help together with molecules specialized in cell adhesion and cell triggering. PMID- 3049120 TI - Microfilament association of ASGP-2, the concanavalin A-binding glycoprotein of the cell-surface sialomucin complex of 13,762 rat mammary ascites tumor cells. AB - Microfilament-associated proteins and membrane-microfilament interactions are being investigated in microvilli isolated from 13,762 rat mammary ascites tumor cells. "Phalloidin shift" analyses on velocity sedimentation gradients of Triton X-100 extracts of [3H]-glucosamine-labeled microvilli identified a 120-kDa cell surface glycoprotein associated with the microvillar microfilament core. The identification was verified by concanavalin A (Con A) blots of one- and two dimensional (2D) electrophoresis gels of sedimented microfilament cores. By 2D electrophoresis and lectin analyses the 120-kDa protein appeared to be a fraction of ASGP-2, the major Con A-binding glycoprotein of the sialomucin complex of the 13,762 cells. This identity was confirmed by immunoblot analyses using immunoblot purified anti-ASGP-2 from anti-membrane serum prepared against microvillar membranes. Proteolysis of the microvilli with subtilisin or trypsin resulted in an increase in the amount of ASGP-2 associated with the microfilament cores. An increase was also observed with sialidase treatment of the microvilli, suggesting that negative charges, probably present on the highly sialated sialomucin ASGP-1 of the ASGP-1/ASGP-2 sialomucin complex, reduce ASGP-2 association with the microfilament core. Proteolysis of isolated microvillar membranes, which contain actin but not microfilaments, also increased the association of ASGP-2 with a Triton-insoluble, actin-containing membrane fraction. Purified ASGP-2 does not bind to microfilaments in sedimentation assays. Since the Triton-insoluble membrane residue is enriched in an actin-containing transmembrane complex, which contains a different glycoprotein, we suggest that the ASGP-2 is binding indirectly via this complex to the microfilament core in the intact microvilli. PMID- 3049122 TI - Extracellular matrix specificity for the differentiation of capillary endothelial cells. AB - Capillary endothelial cells in a fenestrated vasculature contain openings in attenuated areas of the cytoplasm which are often covered with one or two diaphragms. However, due to endothelial cell dedifferentiation upon culturing, such indicative membrane structure are often not expressed. The expression of a more differentiated phenotype can be regulated by the extracellular matrix to which the cells attach. In addition, during angiogenesis endothelial cell migration enables their interaction with other cell types and matrices which may directly affect endothelial cell growth and function. We report the ability to modify the expression of diaphragmed fenestrations and transcapillary channels in capillary endothelial cells by specific extracellular matrices. When the number of membrane openings is measured, growth of the cells on Madin--Darby canine kidney cell matrix results in the greatest number while other matrices or isolated matrix components are only partially active. Production of such a biologically active matrix not only allows an avenue to identify fenestrae associated proteins but also provides an easy culture method to more closely mimic the in vivo phenotype of endothelial cells. PMID- 3049121 TI - Electroporation of cultured adult rat hepatocytes with the c-myc gene potentiates DNA synthesis in response to epidermal growth factor. AB - The human c-myc gene was introduced and transiently expressed in adult rat hepatocyte cultures by the technique of electroporation and its effect on DNA synthesis was examined. Epidermal growth factor (EGF) has been found to stimulate a wave of DNA synthesis in electroporated rat hepatocytes. Hepatocyte cultures electroporated with the c-myc gene showed a potentiation of this EGF effect exhibiting rates of DNA synthesis up to 50% greater than those of control electroporated cultures, as determined by [3H]thymidine labeling of cell nuclei. This potentiation was dependent on the amount of c-myc DNA transfected. The potentiation was due neither to an alteration in the dose-response of the stimulatory effect of EGF nor to a change in the time course of the DNA synthesis wave. PMID- 3049124 TI - Identification and definition of nucleolus-related fibrillar bodies in micronucleated cells. AB - Small nucleolus-related bodies which occur in the nucleoplasm of "micronuclei" lacking nucleolar organizers have been studied by immunofluorescence microscopy. These bodies stained specifically with three different antibodies directed against proteins that are normally associated with the dense fibrillar component of functional nucleoli, but not with antibodies specific for certain proteins of the granular component or the fibrillar centers. Our data show that, in the absence of rRNA genes, the various constituent proteins characteristic of the dense fibrillar component spontaneously assemble into spherical entities but that the subsequent fusion of these bodies into larger structures is prevented in these micronuclei. The similarity between these nucleolus-related bodies of micronuclei and the prenucleolar bodies characteristic of early stages of nucleologenesis during mitotic telophase is discussed. PMID- 3049123 TI - An established rat cell line expressing chondrocyte properties. AB - Chondrocytes express a well-characterized set of marker proteins making these cells useful for studies on differentiation and regulation of gene expression. Because of the inherent instability of primary rat chondrocytes in culture, and because several rat chondrocyte genes have been cloned and characterized (including the collagen II promoter and enhancer), a rat chondrocyte cell line would be especially useful. To obtain this line we infected primary fetal rat costal chondrocytes with a recombinant retrovirus (NIH/J-2) carrying the myc and raf oncogenes, which have been shown to have an "immortalizing" function. Following infection, a rapidly proliferating clonal line was isolated that maintained a stable phenotype through 45 passages (11/2 year in culture). This line, termed IRC, grows in suspension culture as multicellular aggregates and in monolayer culture as polygonal cells which accumulate an alcian blue-stainable matrix. IRC cells synthesize high levels of cartilage proteoglycan core protein, and link protein, but show reduced collagen II expression. In addition, the cells express virally derived myc mRNA and protein, but do not express v-raf. Retinoic acid, which is a known modulator of chondrocyte phenotype, down-regulates expression of chondrocyte marker proteins, while stimulating v-myc expression by IRC cells. These data suggest that v-myc expression by chondrocytes results in rapid cell division and maintenance of many aspects of the differentiated phenotype. These "immortalized" cells, however, remain responsive to agents such as retinoic acid which modulate cell phenotype. The potential exists for development of chondrocyte cell lines from diseased cartilage, as well as from human cartilage. PMID- 3049125 TI - Post-transcriptional regulation by insulin of Xenopus ribosomal protein S6 kinase. AB - The activation of Xenopus oocyte ribosomal protein S6 kinase during oocyte maturation was investigated. Insulin treatment caused a rapid three-fold activation of S6 kinase that returned to near basal levels by 2 h postinsulin. This was followed by a later fivefold increase from 2 to 5 h with insulin, culminating with germinal vesicle breakdown. Pretreatment of oocytes with multiple protein synthesis inhibitors increased the level of basal activity, but did not greatly alter the time course of early activation of S6 kinase by insulin. In contrast, the later increase in S6 kinase activity was completely inhibited by pretreatment with cycloheximide. However, near maximal increases in S6 kinase activity occurred following injection of maturation-promoting factor, even in the presence of multiple protein synthesis inhibitors. Brief exposure to cycloheximide after 30 min or more of insulin stimulation increased the magnitude of insulin-stimulated activity without changing the overall pattern of activity increase. These results suggest that a rapidly turning-over inhibitor of S6 kinase exists, and the activation of S6 kinase by insulin occurs by protein synthesis-dependent and -independent mechanisms. PMID- 3049126 TI - Adhesion of human dermal reticular fibroblasts on complementary fragments of fibronectin: aging in vivo or in vitro. AB - Attachment, spreading, and microfilament reorganization have been evaluated in human dermal reticular fibroblasts isolated from the inner, upper aspect of the arm of a newborn male (RET5 cells) and a 78-year-old male (RET8 cells). Substrata were tested using a set of complementary fragments from individual polypeptide chains of human plasma fibronectin (pFN) or cellular FNs (cFN). With both cell classes, fragments containing the C-terminal heparin-binding (HepII) domain only elicited linear bundles of microfilaments in spreading cells but no stress fibers; fragments containing the RGDS-dependent cell-binding (CellI) domain elicited only partial spreading with condensations of F-actin at ruffling membranes and at other regions along the plasma membrane. The minimum sequence required to obtain responses identical to those on intact pFN (broad spreading with extensive stress fiber formation) was found in fragment 155 (F155) from the beta chain of pFN; F155 contains both HepII and CellI domains. In contrast, the analogous fragment from the alpha chain of pFN (F145) was notably less effective for generating stress fibers. This evidence along with the better attachment, spreading, and microfilament bundle formation on the HepII fragment from the beta chain than the analogous fragment from the alpha chain indicates that the extra type III homology unit permits more effective interaction of beta chain fragments with cell-surface heparan sulfate proteoglycan and possibly integrin (binding efficiency to the substratum was similar for fragments from both chains). Therefore, alternatively spliced sequences that neighbor binding domains can play significant roles in the interaction of the domain with cell-surface receptors of dermal fibroblasts. Comparison of RET5 responses with those of RET8 cells has identified changes in adhesive mechanisms as cells undergo "aging" processes. Attachment and microfilament bundle formation were far more effective for RET5 cells than for RET8 cells on any of the HepII fragments. Conversely, RET8 cells were far more sensitive to an RGDS-containing peptide in their medium on CellI fragments than RET5 cells. These results together indicate that in vivo aging leads to greater dependence upon cell-surface integrin binding and less dependence upon heparan sulfate proteoglycan binding for responses on FN matrices. When RET5 cells entered senescence (in vitro aging), they also became much more sensitive to peptide A. On several fragments and on intact pFN, RET8 cells generated very thick stress fibers that were observed only on one fragment with RET5 cells.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3049127 TI - Trigramin, an RGD-containing peptide from snake venom, inhibits cell-substratum adhesion of human melanoma cells. AB - Trigramin, a cysteine-rich, RGD-containing peptide isolated from the venom of the Trimeresurus gramineus snake, inhibited the adhesion of human melanoma cells to fibronectin and fibrinogen. Compared on a molar basis to GRGDSP, trigramin was approximately 500 times more potent than the hexapeptide at inhibiting cell adhesion to fibronectin. The activity of trigramin was abolished by chemical reduction of the molecule, indicating that the secondary structure is important to the biological activity. Trigramin presents an example of an effective inhibitor of cell adhesion that has developed in nature and may prove to be a useful probe in studying the cell surface receptors involved in cell adhesion. PMID- 3049128 TI - Cell cycle-dependent reactivity with the monoclonal antibody Ki-67 during myeloid cell differentiation. AB - The specificity and sensitivity of the monoclonal antibody Ki-67 in identifying proliferating cell compartments was tested with the human promyelocytic leukemia cell line HL-60 using multi-parameter flow cytometry. While correlated measurements of DNA content and Ki-67 immunofluorescence indicated that the antigen was present in all phases of the cell cycle, reactivity with the antibody was highest in proliferating S and G2+M cells. The analysis of the BrdU content of cells sorted on the basis of reactivity with Ki-67 showed a correlation between Ki-67 reactivity and BrdU uptake. In HL-60 cells induced to differentiate with dimethyl sulfoxide (DMSO), the loss of reactivity with Ki-67 paralleled the exit of cells from the cell cycle. This was not observed in DMSO-resistant HL-60 cells. These results validate the usefulness of the Ki-67 antibody for determining the proliferative stage of mammalian cells in culture. PMID- 3049130 TI - Induction of immune resistance against L1210 lymphatic leukemia in mice after chemoradiotherapy of the leukemia and reconstitution with bone marrow purged from the leukemia with mafosfamide. AB - Lymphatic leukemia L1210-bearing semisyngeneic Balb/c x DBA/2Wf F1 (CD2F1) mice were subjected to chemoradiotherapy (2 x 100 mg/kg of cyclophosphamide i.p. and 1000 cGy of total body irradiation) and reconstitution with 10(7) syngeneic bone marrow cells i.v. The bone marrow obtained from leukemic mice was previously ex vivo purged of the leukemia cells with mafosfamide (ASTA Z7654) and stored in liquid nitrogen. Eight weeks after cytoreductive therapy and bone marrow transplantation we tried to immunize the mice against the lethal dose of the leukemia by i.p. injections of L1210-Maf cells (L1210 cells treated in vitro with mafosfamide for inhibition of their growth). About 75% of such mice were able to reject the subsequent 10(3) L1210 leukemia cell challenge, as compared with 70% of normal immunized mice and 55% of mice reconstituted with bone marrow cells not treated with mafosfamide. PMID- 3049131 TI - Mechanisms contributing to the sex difference in levels of granulocyte-macrophage colony-stimulating factor in the urine of GM-CSF transgenic mice. AB - Levels of granulocyte-macrophage colony-stimulating factor (GM-CSF) were 30- to 40-fold higher in the urine of male GM-CSF transgenic mice than in female transgenic mice, despite uniform elevations in both sexes of serum GM-CSF levels. Male transgenic bladder tissue produced two to four times more GM-CSF in vitro than female transgenic or control bladder tissue, but no sex differences were observed in the production of GM-CSF in vitro by kidney tissue. No sex differences were observed in the serum half-lives of native or recombinant GM-CSF in C57BL or littermate control mice, and the half-lives of recombinant GM-CSF were shorter than those of native GM-CSF. The studies indicated that some GM-CSF in urine can represent plasma GM-CSF cleared by the kidney, and native GM-CSF was cleared to the urine more efficiently than recombinant GM-CSF. Female transgenic mice exhibited a subnormal capacity to clear injected native GM-CSF to the urine. Although granulomas were present in the bladder wall of some transgenic mice, their presence did not correlate with the GM-CSF levels in the urine. PMID- 3049129 TI - Radiosensitivity of cloned permanent murine bone marrow stromal cell lines: nonuniform effect of low dose rate. AB - The x-irradiation biology of supportive stromal cells of the bone marrow microenvironment was investigated by using cloned permanent cell lines that were established from hematopoietically active murine long-term bone marrow cultures. X-irradiation survival curves were derived for each cell line at either 120 cGy/min or clinical low dose rate (LDR) (5 cGy/min) that is used in total body irradiation protocols prior to bone marrow transplantation. Four cell lines, MBA 1, MBA-13, 14F2.1, and D2XRII (Group I) demonstrated a significant increase in D0 at 5 cGy/min (280 cGy, 270 cGy, 210 cGy, and 240 cGy, respectively) compared to 120 cGy/min (215 cGy, 210 cGy, 157 cGy, and 210 cGy, respectively) (p less than 0.05). In contrast, three other clonal cell lines, +/+ #2 cl 4, S1d #3, and GPI alpha-1 (Group II) showed no significant dose rate dependent change in D0 at 5 cGy/min (164 cGy, 174 cGy, and 159 cGy, respectively) compared to 120 cGy/min (159 cGy, 167 cGy, and 143 cGy, respectively). Group I and II cell lines could not be distinguished by differences in synthesis of extracellular matrix proteins including laminin, collagen types I and IV, or histochemically detectable enzymes. Three Group I lines (MBA-1, MBA-13, and D2XRII) and one Group II line, Sld #3, showed decreased support capacity for cocultivated hematopoietic stem cells in vitro. All seven lines had detectable polyA+ mRNA for monocyte colony stimulating factor (M-CSF) as detected by molecular hybridization; one had detectable levels of polyA+ mRNA for granulocyte-macrophage colony-stimulating factor (GM-CSF) (D2XRII); one, detectable levels of polyA+ mRNA for interleukin 1 (IL-1); and none of the seven had detectable polyA+ mRNA for granulocyte colony stimulating factor (G-CSF) or IL-3 (multi-CSF). The data indicate that some cells of the hematopoietic microenvironment may not be selectively protected by clinical low dose rate irradiation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3049132 TI - Radioprotection of hemopoiesis conferred by Acanthopanax senticosus Harms (Shigoka) administered before or after irradiation. AB - Acanthopanax senticosus Harms (Shigoka) extract has been observed to have radioprotective effects on hemopoiesis of irradiated mice (CBA/olac) when administered before or after irradiation by 60Co. The mechanisms of action were explored by studying the following parameters: 1) survival after lethal doses of irradiation; 2) recovery of spleen colony-forming units (CFU-S); 3) protective effect on endogenous CFU-S; and 4) effect on self-renewal of CFU-S. 1) The 30-day mortality due to bone marrow failure after irradiation was significantly reduced in Shigoka-treated mice. The maximum effect of the extract (80% survival) at a dose of 5 mg was observed when given intraperitoneally 24 h before a lethal dose of irradiation (9.5 Gy) and it was still effective when administered as late as 12 h after irradiation (30% survival). The radioprotective effect of Shigoka extract given before irradiation was not due to an increase in the total number of CFU-S but probably due to quiescent CFU-S that entered DNA synthesis (S-phase) 24 h after the injection of Shigoka extract. 2) Recovery of CFU-S in mice given the extract within 15 min after 4.75 Gy of whole body irradiation was also enhanced, especially in the spleen. The number of CFU-S in the bone marrow 24 h after irradiation showed a marked decrease. It returned to normal values in the bone marrow at day 11 in Shigoka-treated mice and at day 15 in nontreated irradiated mice. CFU-S in the spleen recovered more rapidly with an overshoot from days 7 to 21 in the Shigoka-treated mice, whereas in control irradiated mice it did not reach the normal value until day 21. 3) Nine days after sublethal doses of irradiation (7.0 Gy) the number of endogenous spleen colonies (endogenous CFU-S) was highest in mice given the extract 24 h before irradiation. The extract administered as late as 24 h after irradiation also resulted in an increase of the endogenous CFU-S. 4) The number of CFU-S in each 9-day endogenous CFU-S was 73.9 +/- 7.2 per nodule in treated mice and 0.2 +/- 0.1 per nodule in irradiated control mice, which suggests increased self-renewal of CFU-S in Shigoka-treated groups. These results suggest that the radioprotection conferred by Shigoka extract results from enhanced stimulation of CFU-S not only toward proliferation but also toward CFU-S self-renewal. These two phenomena could explain the protective effects of Shigoka extract administered even after irradiation. PMID- 3049133 TI - Enhanced sprouting of retinotectal fibers after early superior colliculus lesions in hamsters treated with gangliosides. AB - The effects of exogenous gangliosides on sprouting of optic tract axons was studied in hamsters which, after a right tectal lesion on the day after birth (P1), had an abnormal retinotectal projection from the left eye to the left superior colliculus (SC). Sprouting of these axons was induced by removing the competing input by right eye removal on postnatal day 9 (P9). Intraperitoneal GM1, given daily and started on P9, significantly stimulated the sprouting response. This was demonstrated by Fink-Heimer silver staining of anterograde axonal degeneration three days after the left eye was removed on P36. Terminal fields in the left SC were, in average, twice as large compared to controls. An estimate of the total number of terminals (silver stained particles) revealed a value of 7.9 X 10(6) for GM1 and 3.2 X 10(6) for control hamsters, respectively. Diencephalic structures which also receive collateral input from the sprouting optic tract did not show any alterations in the size of the terminal field due to GM1-treatment, suggesting that, in vivo, gangliosides fail to initiate sprouting in areas that have not previously been denervated. Unexpectedly, GM1-treated hamsters also had significantly smaller right SC damage and less left damage near the midline. Subsequent reanalysis of the data based on a lesion-matching procedure indicates that effects on reducing atrophy were independent of the GM1 enhanced sprouting of retinofugal axons. These findings provide the first direct evidence that exogenous GM1 stimulates lesion-induced axon sprouting in the mammalian brain. PMID- 3049135 TI - Brugia malayi: detection of parasite antigen in sera from infected jirds. AB - Sera from Brugia malayi-infected jirds were demonstrated to contain a heat stable, 95- to 105-kDa parasite antigen by immunoblot with rabbit antibody to the parasite and with a monoclonal antibody that binds to phosphorylcholine. This antigen is a major component of B. malayi adult worm excretory/secretory antigen, and it is present in lavage fluid obtained from ip-infected animals. The antigen was detected by enzyme immunoassay in all sera collected from jirds 9-54 weeks after sc injection with 100 or 300 infective larvae (L3). Parasite antigen titers were higher in animals infected with the higher L3 dose. Antiphosphorylcholine antibodies were present in jird sera for the first 12 weeks after larval injection, but thereafter, antibody titers decreased to undetectable levels. Parasite antigen was not detected by immunoblot or enzyme immunoassay in sera from 21 human subjects with B. malayi microfilaremia. Antigen may be cleared from human sera by antiphosphorylcholine antibodies, which were present in all sera tested. The practical significance of B. malayi antigen detection in the jird is that it provides a sensitive means of noninvasively monitoring the status of infection in this important experimental filariasis model. PMID- 3049134 TI - Plasmodium falciparum: gene structure and hydropathy profile of the major merozoite surface antigen (gp195) of the Uganda-Palo Alto isolate. AB - The gene encoding the 195,000-Da major merozoite surface antigen (gp195) of the FUP (Uganda-Palo Alto) isolate of Plasmodium falciparum, a strain widely used for monkey vaccination experiments, has been cloned and sequenced. The translated amino acid sequence of the FUP gp195 protein is closely related to the sequences of corresponding proteins of the CAMP (Malaysia) and MAD-20 (Papua New Guinea) isolates and more distantly related to those of the Wellcome (West Africa) and K1 (Thailand) isolates, supporting the proposed allelic dimorphism of gp195 within the parasite population. The prevalence of dimorphic sequences within the gp195 protein suggests that many gp195 epitopes would be group-specific. Despite the extensive differences in amino acid sequence between gp195 proteins of these two groups, the hydropathy profiles of proteins representative of both groups are very similar. The conservation of overall secondary structure shown by the hydropathy profile comparison indicates that gp195 proteins of the various P. falciparum isolates are functionally equivalent. This information on the primary structure of the FUP gp195 protein will enable us to evaluate the possible roles of conserved, group-specific and variable epitopes in immunity to the blood stage of the malaria parasite. PMID- 3049137 TI - Respiratory events during sleep Amiens: 19th-20th November, 1987. PMID- 3049138 TI - Bronchial reactivity in the community. PMID- 3049136 TI - Objective measurement of compliance with nasal CPAP treatment for obstructive sleep apnoea syndrome. AB - Compliance with nasal continuous positive airway pressure (CPAP) has become a major concern, since this treatment is efficacious, but constraining. In 46 consecutive obstructive sleep apnoea (OSA) patients, we measured compliance with nasal CPAP by establishing a mean rate of use, with a built-in time counter read at three-month intervals, over a mean follow-up period of 232 +/- 27 days. The mean rate of use in the whole group was 5.14 +/- 0.31 hours per day. The acceptance rate was 90.9-93.2%, showing that patient acceptance is not a limitation in the use of nasal CPAP. PMID- 3049139 TI - Is hyperreactivity the same as asthma? PMID- 3049140 TI - Spontaneous changes of airway hyperresponsiveness in bronchial asthma. PMID- 3049141 TI - Methods of testing nonspecific bronchial hyperresponsiveness. PMID- 3049142 TI - The relationship between reversibility and hyperreactivity. PMID- 3049143 TI - Bronchial hyperreactivity in smokers. PMID- 3049144 TI - Mucosal inflammation and bronchial hyperreactivity. PMID- 3049145 TI - The significance of bronchial responsiveness in children. PMID- 3049146 TI - Measuring health status in chronic airflow limitation. AB - A health status instrument for use in clinical trials must be valid (measuring what it is supposed to measure) and responsive (able to detect clinically important change). Approaches to measuring health status in clinical trials include using a battery of instruments, a general instrument which provides a profile of the patient's health, an instrument that generates a health utility, or an instrument that focuses on the problems associated with a particular disease. Disease-specific instruments have been used in clinical trials in chronic airflow limitation (CAL). The Oxygen Cost Diagram is simple and easy to administer, but responsiveness and validity are unproven. The Transition Dyspnoea Index is valid and responsive, but is difficult to use in trials in which multiple measurements are desired. The Chronic Respiratory Disease Questionnaire has proved valid and responsive in controlled trials in CAL patients. Health status measures should be included in all clinical trials in CAL. PMID- 3049147 TI - A cDNA clone encoding a 10.8 kDa photosystem I polypeptide of barley. AB - A cDNA clone encoding the barley photosystem I polypeptide which migrates with an apparent molecular mass of 16 kDa on SDS-polyacrylamide gels has been isolated. The 634 bp sequence of this clone has been determined and contains one large open reading frame coding for a 15,457 Da precursor polypeptide. The molecular mass of the mature polypeptide is 10,821 Da. The amino acid sequence of the transit peptide indicates that the polypeptide is routed towards the stroma side of the thylakoid membrane. The hydropathy plot of the polypeptide shows no membrane spanning regions. PMID- 3049148 TI - Globo-A--a new receptor specificity for attaching Escherichia coli. AB - Uropathogenic Escherichia coli strains designated as ONAP, based on their O negative A positive agglutination of human P1 erythrocytes, were shown to prefer the globo-A glycolipid as a receptor structure. The dependence on both the A terminal and the globoseries chain was confirmed by agglutination of human AP1, but not Ap or OP1 erythrocytes and by binding to the globo-A glycolipid on TLC plates. Neither Gal alpha 1----4Gal beta nor the A trisaccharide GalNAc alpha 1-- -3(Fuc alpha 1----2)Gal beta alone functioned as receptors. The bacteria thus appeared to recognize an epitope resulting from the combination of the terminal and internal structures. PMID- 3049149 TI - Differential sensitivity to pertussis toxin of 3T3 cells transformed with different oncogenes. AB - Pertussis toxin (PT), which blocks the activity of several G-proteins, has been found to exert a marked inhibitory effect on the DNA synthesis induced in 3T3 cells by serum or growth factors. 3T3 cells transformed with human c-ras oncogenes (Ha-ras, Ki-ras, N-ras) or with src, an oncogene coding for a protein kinase, have lost sensitivity to growth control by PT, even though substrates for PT can still be ADP-ribosylated in vivo. In contrast, 3T3 cells transformed with the SV40 virus behave like normal untransformed cells with respect to the ability of PT to decrease their growth rate. Oncogenes can thus likely be classified either as 'responders' or 'non-responders' to PT. PMID- 3049150 TI - Primary and post-irradiation inactivation of the sulfhydryl enzyme malate synthase: correlation of protective effects of additives. AB - The presence of additives during X-irradiation of malate synthase led to radioprotective effects against primary and post-irradiation inactivation. Pronounced effects were provided by typical scavengers, sulfhydryl reagents and specific ligands (substrates, products, analogues). The results show that scavenging and specific protection are responsible for the protective efficiency of additives. Scavengers delete noxious species formed during irradiation or post radiationem. Sulfhydryl reagents may act as repair substances. Specific ligands protect the active site of the enzyme and the essential sulfhydryls; specific protection is more pronounced post-radiationem. Ligands and sulfhydryl reagents may additionally act as scavengers. A cumulative index for the protective power of additives against both sorts of inactivation was established. PMID- 3049151 TI - Expression of the glyceraldehyde-3-phosphate dehydrogenase gene from the extremely thermophilic archaebacterium Methanothermus fervidus in E. coli. Enzyme purification, crystallization, and preliminary crystal data. AB - The gene of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from the extremely thermophilic archaebacterium Methanothermus fervidus (growth optimum 82 degrees C) was cloned in vector pJF118EH and expressed in E. coli cells. As shown by molecular mass determination, protein sequencing, heat stability, and substrate saturation kinetics, the enzyme synthesized in E. coli is identical to the original enzyme from M. fervidus. The high thermostability of the E. coli produced M. fervidus GAPDH allows rapid purification to homogeneity. From this enzyme protein crystals were grown which proved to be suitable for X-ray analysis. The crystals are of tetragonal space group P4(1)22 and contain a dimer per asymmetric unit. PMID- 3049152 TI - Nucleotide sequence of cDNA clones encoding the entire precursor polypeptides for subunits IV and V of the photosystem I reaction center from spinach. AB - Using lambda gt11 expression cloning and immunoscreening, cDNA-containing recombinant phages for subunits IV and V of the photosystem I reaction center were isolated, sequenced and used to probe Northern blots of polyadenylated RNA prepared from spinach seedlings. The mRNA sizes for both components are approximately 1000 and 850 nucleotides, respectively. The 968 nucleotide cDNA sequence and derived amino acid sequence for subunit IV predict a single open reading frame of 231 amino acid residues (25.4 kDa). Comparison with a 13-residue N-terminal amino acid sequence determined for subunit IV suggests a mature protein of 17.3 kDa (154 residues) and a transit sequence of 77 amino acids (8.1 kDa). The corresponding data for subunit V are 677 bp (cDNA), 167 residues for the precursor protein (18.2 kDa), 98 residues for the mature polypeptide (10.8 kDa) and 69 residues for the transit peptide (7.4 kDa). Secondary structure predictions indicate that both proteins possess greatly different transit sequences and that none is membrane-spanning. PMID- 3049154 TI - Mutagenesis directed by phosphotriester analogues of oligonucleotides: a way to site-specific mutagenesis in vivo. AB - A new approach to induce directed mutations in genes of study through simple cotransfection of E. coli cells by the mixture of primer and template was developed. This method is based on the use of synthetic phosphotriester analogues of oligonucleotides as site-specific mutagenic primers. The achieved yield of mutant clones was 2-3%. PMID- 3049153 TI - The cDNA clone for strictosidine synthase from Rauvolfia serpentina. DNA sequence determination and expression in Escherichia coli. AB - The cDNA clone for strictosidine synthase, the enzyme which catalyzes the stereospecific condensation of tryptamine with secologanin to form the key intermediate in indole alkaloid biosynthesis, strictosidine, has been identified with a synthetic oligodeoxynucleotide hybridization probe in a lambda gt11 cDNA library of cultured cells of Rauvolfia serpentina. The DNA has been sequenced, revealing an open reading frame of 1032 base pairs encoding 344 amino acids. The sequence of 60 nucleotides in the 5'-flanking region has been determined by primer extension analysis. The encoded protein has been expressed in E. coli DH5 as detected by immunoblotting of protein extracts with antibodies raised against the native enzyme. PMID- 3049155 TI - Total synthesis of the cystatin alpha gene and its expression in E. coli. AB - A gene encoding cystatin alpha has been chemically synthesized, cloned and expressed in E. coli. The gene of 318 base pairs was assembled by enzymatic ligation of 19 oligonucleotides and cloned into a pBR322-derived expression plasmid down stream of the tac promoter. The expression product of the synthetic gene has been purified by Sephadex G-50 column chromatography and shown to have the same properties as those of the authentic protein isolated from rat epidermis. PMID- 3049156 TI - Long-range 15N-1H correlation as an aid to sequential proton resonance assignment of proteins. Application to the DNA-binding protein ner from phage Mu. AB - A method is described for sequential resonance assignment of protein 1H-NMR spectra relying on the detection of long-range correlations between 15N and C alpha H atoms using 1H-detected heteronuclear multiple-bond correlation spectroscopy. In particular, the observation of the two-bond 15N(i)-C alpha H(i) and three-bond 15N(i)-C alpha H(i-1) correlations enables one to connect one residue with the next. Because the magnitude of the long-range couplings is small (less than 6 Hz), the sensitivity of this experiment is necessarily low and requires the use of 15N-enriched protein samples. Further, because the size of the 15N(i)-C alpha H(i-1) coupling is very sensitive to the psi backbone torsion angle, structural information can be derived. The application of this experiment is illustrated with the 75-residue DNA-binding protein ner from phage Mu. PMID- 3049157 TI - tRNAPhe deprived of 3'-terminal adenosyl residue does not stimulate adenosine aminoacylation catalyzed by phenylalanyl-tRNA synthetase from Escherichia coli. AB - Phenylalanyl-tRNA synthetase from Escherichia coli does not catalyze the [14C]phenylalanyl residue transfer from phenylalanyl-adenylate to adenosine either in the presence or absence of homologous tRNAPhe and tRNA(-A Phe). When the reaction mixture contained dithiothreitol, radioactive substance was detected having a mobility on HPLC column close to that of aminoacyladenosine. The amount of this product depended on the concentration of dithiothreitol in the mixture. Phenylalanyl residue was suggested to undergo transfer from aminoacyladenylate to dithiothreitol molecule. PMID- 3049158 TI - Location of divalent ion sites in acyl carrier protein using relaxation perturbed 2D NMR. AB - The T1-accordion COSY experiment has been applied to acyl carrier protein (ACP) to locate the divalent ion binding sites in the protein using the paramagnetic ion, Mn2+, as a substitute for Ca2+. Replacement with Mn2+ leads to an enhancement of proton spin-lattice (T1) relaxation rates. These enhancements have a 1/r6 distance dependence that makes them extremely useful in structural analyses. Ion-proton distances ranging from 3.0 to 9.0 A have been obtained from this experiment and subsequently used as constraints in the molecular mechanics module of AMBER to refine a protein structure. PMID- 3049159 TI - The 'molten globule' state is involved in the translocation of proteins across membranes? AB - Strong evidence exists that the translocation of proteins across a variety of membranes involves a non-native or denatured conformational states. On the other hand a compact state having secondary but not rigid tertiary structure and called the 'molten globule' state has been identified as being stable under mild denaturing conditions. A similar state has been shown to accumulate on the folding pathway of globular proteins. These states are compact though sufficiently expanded to include water, and they are internally mobile. It is proposed that these molten globule states may be suitable candidates for protein translocation across biological membranes. PMID- 3049161 TI - Alterations in the cleavage site of the signal sequence for the secretion of human lysozyme by Saccharomyces cerevisiae. AB - The amino acids corresponding to the cleavage site of a hybrid preprotein containing a chicken lysozyme signal and a mature portion of human lysozyme were altered. The processing of mutant signals of -3Pro and -3Asp/-1Ala decreased remarkably, while that of -2Pro was 75% of that of the native signal. The major cleavage site of -3Pro was the same as that of the native signal, but that of the -2Pro and -3Asp/-1Ala signals was shifted one residue closer to the N-terminal side than the original site. The cleavage of the -2Pro signal, which was identical to the native processing of pheasant prelysozyme, suggested that the signal peptidases in yeast and bird are similar. PMID- 3049160 TI - Cloning and high-level expression of a chloroperoxidase gene from Pseudomonas pyrrocinia in Escherichia coli. AB - A chloroperoxidase gene from Pseudomonas pyrrocinia was cloned into Escherichia coli using the cosmid vector pJB8. The gene coding for the chloroperoxidase could be localized to a 1.5 kb fragment of DNA which was subcloned into the high-copy number plasmid pUC18. In one subclone increased halogenating activity could be found which was 570-fold greater than in P. pyrrocinia. The halogenating enzyme was identified as the chloroperoxidase by SDS-polyacrylamide gel electrophoresis. PMID- 3049162 TI - Two conserved tryptophan residues of tumor necrosis factor and lymphotoxin are not involved in the biological activity. AB - Each of the two highly conserved tryptophan residues in hTNF (positions 28 and 114) was converted into phenylalanine by site-directed mutagenesis and the mutant proteins were partially purified. A cytotoxicity assay on mouse L929 cells showed only a slight reduction in biological activity, strongly suggesting that neither of the two amino acids is involved in the active site. PMID- 3049163 TI - The herbicidally active experimental compound Hoe 704 is a potent inhibitor of the enzyme acetolactate reductoisomerase. AB - Growth inhibition of plants and bacteria by the experimental herbicide Hoe 704 (2 methylphosphinoyl-2-hydroxyacetic acid) was alleviated by the addition of the branched-chain amino acids to growth media. Hoe 704 caused a massive accumulation of acetoin and acetolactate, indicating its direct interference with the branched chain amino acid biosynthetic pathway. The second enzyme of this pathway, acetolactate reductoisomerase (EC 1.1.1.86), was found to be subject to strong inhibition by Hoe 704. The inhibition was time-dependent and competitive with the enzyme's substrate, acetolactate. This report establishes acetolactate reductoisomerase as a new target for a herbicidal compound. PMID- 3049164 TI - A community screening programme for abdominal aortic aneurysms. AB - Three-hundred and sixty-nine unselected men aged 65-79 years were invited for screening for abdominal aortic aneurysm. One hundred and forty-one men were examined and 4 aneurysms detected. 43.2% of men aged 65-74 attended for examination in response to a single unsolicited letter of explanation with the date of an appointment, but only 29.1% of those aged 75-79 years. It is suggested that community mortality from ruptured aortic aneurysm could be reduced by ultrasound screening of the aorta in men aged 65-74 years and early selective aneurysm surgery. PMID- 3049165 TI - Current status of biotechnological studies in mammalian reproduction. PMID- 3049167 TI - Capacity of objectively assessed sperm motility characteristics in differentiating between semen of fertile and subfertile men. AB - A simple and inexpensive computer-assisted method for the objective assessment of sperm motility characteristics was employed to evaluate semen samples of 42 fertile men and 70 subfertile patients. The capacity of each motility parameter to discriminate between semen of the two groups was evaluated by receiver operating characteristic curve analysis. Velocity, linear velocity, and "angular" velocity are reasonably accurate, whereas linearity and angularity index had a very poor discriminating power. The best discrimination between the two groups was the proportion and concentration of sperm with rapid linear progressive motility, based on the cut-off value of linear velocity greater than or equal to 22 microns/sec. This parameter was 90% accurate in discriminating semen of infertile men from that of subfertile patients. PMID- 3049166 TI - Contragestion with late luteal administration of RU 486 (Mifepristone). AB - The efficacy and tolerance of RU 486 prescribed as a late luteal contragestive agent have been evaluated in 139 women at risk of pregnancy. They were given 400 or 600 mg of RU 486 once on the day before the expected menses. Among these women, 48 (34.5%) were pregnant (positive plasma beta-human chorionic gonadotropin, [beta-hCG]) at the time of RU 486 intake. Bleeding occurred in all but six women. An ongoing pregnancy after treatment was found in nine cases (failure rate, 9/48, 18.8%), which was subsequently terminated by surgical procedure in all cases. There was no disturbance in the menstrual cycle, and the tolerance was very satisfactory. In conclusion, this method is acceptable for women at risk of pregnancy in whom other usual postcoital contraceptive methods cannot be prescribed. PMID- 3049168 TI - Orthotopic tibia transplantation in mice. PMID- 3049169 TI - [Inflammatory connective tissue diseases--recent knowledge for diagnosis and therapy in general practice]. PMID- 3049170 TI - [Results of therapy studies with anti-inflammatory substances in occlusive conditions in psoriasis]. PMID- 3049171 TI - [Johann Wilhelm Ritter--the discoverer of ultraviolet radiation]. PMID- 3049172 TI - [Specific nuclear binding pattern of the scleroderma-70 antibody in immunofluorescence detection of antinuclear factors]. PMID- 3049173 TI - Fenticonazole cream once daily in dermatomycosis, a double-blind controlled trial versus bifonazole. AB - A randomized, double-blind clinical trial was undertaken in 41 patients with dermatomycosis to compare topical 2% fenticonazole cream (group A: 21 patients) with topical 1% bifonazole cream (group B: 20 patients). Treatment was performed as a once daily application. Mycological and clinical parameters were assessed before treatment and after 7, 14, 21 and 28 days. At the control visits the clinical investigator also expressed an overall judgement on the patient's state of disease. This parameter was based on a combined clinical and mycological assessment by the physician; laboratory screening investigations were undertaken before and at the end of treatments. All patients were checked for their state of disease 3-4 weeks after the end of treatment. All assessment criteria showed fenticonazole to be at least as efficacious as bifonazole. Several trends in favor of fenticonazole were also found: fenticonazole achieved superior results in the overall clinical evaluation, and after 3 weeks of treatment 15 patients out of 21 on fenticonazole were cured in mycological and clinical terms, whereas treatment with bifonazole resulted in complete healing of only 7 patients out of 20. This difference is statistically significant (p = 0.021) and indicates a more rapid therapeutic activity of fenticonazole. At the posttreatment rechecks no recurrent disease was registered, irrespective of whether patients had received fenticonazole or bifonazole. Laboratory screening investigations revealed no evidence of significant treatment-related changes or abnormalities in both treatments. No adverse events were noted for either treatment. PMID- 3049175 TI - Corticosteroids and immune systems of non-mammalian vertebrates: a review. PMID- 3049174 TI - Thymopentin in the treatment of severe alopecia areata. AB - Since alopecia areata might be due to an aberrant T-cell-mediated immunity, the purpose of our study was to compare the results obtained with Thymopentin to those of topical sensitizing therapies, such as squaric acid dibutylester or diphencyprone. Statistical analysis showed no differences between the results obtained with these two therapies. Therefore, Thymopentin can be considered to be a new therapeutic agent which, like the classic topical drugs squaric acid dibutylester or diphencyprone, has immunoreactive properties and might provide some hope to patients with severe alopecia areata. PMID- 3049176 TI - Doppler ultrasound of the uteroplacental circulation as a screening test for severe pre-eclampsia with intra-uterine growth retardation. AB - Two hundred primiparae underwent continuous-wave Doppler investigation of the uteroplacental circulation at 18-20 weeks gestation as a possible screening test for hypertension in pregnancy. Seventy-five women with abnormal waveforms suggestive of high uteroplacental resistance were tested again at 24 weeks when 21 demonstrated a persistent abnormality. Only nine (43%) of these went on to have an uncomplicated pregnancy, as compared with 150 (84%) of the remainder. Seventeen (8.5%) of the women in the study developed a hypertensive disorder of pregnancy, five of whom had abnormal waveforms at 18-20 weeks and at 24 weeks. These five women had a more severe degree of hypertension with proteinuria or intra-uterine growth retardation, and two required clinical intervention before term. The remaining 12 women were delivered at term of average, or heavier than average babies. Doppler investigation of the uteroplacental circulation at 24 weeks may prove to be a sensitive screening test for later severe pre-eclampsia with intra-uterine growth retardation. PMID- 3049177 TI - Prediction and self-prediction of ovulation in clomiphene citrate-treated patients. AB - We studied 15 infertile patients for a total of 25 cycles in order to compare the predictive value of the following parameters in timing ovulation: basal body temperature (BBT), cervical mucus, serum E2 and LH, follicular growth as shown by ultrasonography, and urinary LH. The patients themselves tested urinary LH at home using Clearplan kits. BBT had low value as a predictive test for the time of ovulation, whereas ultrasonography had a predictive value of 9%, serum E2 50%, cervical mucus 59%, serum LH 63.6% and Clearplan kits 63.6%. The best predictive reliability in timing ovulation was obtained by considering both serum LH and E2 measurements. The Clearplan kits were as reliable as serum LH measurements and have greater practical advantages. PMID- 3049178 TI - Colicin E1 in planar lipid bilayers. AB - The channel formed by the C-terminal domain of colicin E1 in planar lipid bilayers has proven to be more complex than one might have guessed for such a simple system. The protein undergoes a pH-dependent rearrangement which transforms it from a water soluble form to a much different membrane bound form. There are at least two bound states which don't form a channel. The process by which the channel opens and closes is regulated by the pH and the transmembrane voltage. The voltage is probably sensed by at least 3 (and more likely 4 or more) lysine residues which must be driven through the field to open the channel. The process appears to be hindered by particular carboxyl groups when they are in the unprotonated state. The open channel has several substates and several superstates. Very large positive voltage catalyzes a transition of the open channel to an inactivated state, and may be able to drive the channel-forming region of the protein across the membrane. Little is known about the structure of any of these states, but the open channel is large enough to allow NAD to traverse the membrane and appears to be formed by one colicin molecule. This single polypeptide mimics many of the properties found in channels of mammalian cell membranes, but it may prove more relevant as a model for the transport of proteins across membranes. The comparative ease with which the protein can be manipulated chemically and genetically, along with the complexity of its behavior, promises to keep several laboratories busy for some time. PMID- 3049180 TI - Regulation of skeletal muscle myofibrillar protein degradation: relationships to fatigue and exercise. AB - 1. Exercise results in large alterations in cellular metabolic homeostasis and protein turnovers. Exhaustive exercise (as well as starvation, dystrophy, motor nerve disease) results in myofibrillar degradation and has been associated with the decreased force generating capabilities of muscle at fatigue. 2. Complete protein degradation is accomplished by the combined actions of non-lysosomal and lysosomal proteases and the initial breakdown of myofibrillar protein appears to be non-lysosomal mediated. 3. Current evidence suggests that covalent modification (mixed-function oxidation, formation of mixed disulfides, oxidation of methionine residues and phosphorylation) of proteins may mark them for degradation by rendering them more susceptible to proteolytic attack. 4. The rate of covalent modification can be controlled by the level of stabilizing and destabilizing ligands and by factors affecting the activity of the marking reaction. 5. The activities of individual proteases may be controlled by activators and inhibitors. 6. It is suggested that the large alterations in metabolism (hormonal profiles, energy status, redox status and Ca2+ levels) which accompany exercise serve to activate specific proteases and/or induce covalent modifications which mark specific myofibrillar proteins for subsequent proteolytic attack. PMID- 3049179 TI - Radioiodinated monoclonal antibodies. PMID- 3049181 TI - Compared roles of glucose, galactose and fructose as glycogen precursors during the acute response to insulin in cultured rat foetal hepatocytes. AB - 1. The efficiency of the contribution of hexoses to basal- and stimulated glycogenesis, when studied in cultured 18 day-old rat foetal hepatocytes in the presence of glucose, was as follows: galactose greater than glucose greater than fructose. 2. Glucose deprivation had opposite effects on the contributions of [14C]galactose (decreased) and [14C]fructose (increased) to glycogenesis, which occurred independently of insulin and were reversed by glucose concentrations as low as 30-100 microM. 3. The stimulation of glycogenesis by insulin measured with [14C]glucose (3.2-fold) was superior to that obtained with either [14C]galactose or [14C]fructose (2.7-fold in both cases), which revealed a specific beneficial effect of insulin on glucose contribution. PMID- 3049182 TI - Changes in lysosome populations in the rat kidney cortex induced by experimental proteinuria. AB - 1. Experimental proteinuria (262.9 mg protein/24 hr urine) was induced in rats by repeated intraperitoneal injections of BSA. 2. Hypertrophy of the kidney cortex was significant 8 days after the start of the BSA injections, and the activities of lysosomal enzymes in kidney cortex and urine were significantly higher in proteinuric compared to nonproteinuric rats. 3. Lysosome populations in the kidney cortex were examined by rate sedimentation of the homogenate and by rate zonal and isopycnic centrifugation of the lysosome-rich ML fraction. 4. The activity of lysosomal enzymes in the kidney cortex increased slightly, essentially in the large, fragile lysosomes mainly recovered from the proximal tubule. 5. Proteinuria induced a shift/reduction in the density of small lysosomes from 1.235 and 1.20 g/ml to 1.225 and 1.185 g/ml, respectively. 6. Proteinuria induced a new population of small lysosomes (density 1.185 g/ml) enriched in cathepsin D. PMID- 3049183 TI - Proteolytic activity in electropherograms (SDS-PAGE) of bovine erythrocyte ghosts. AB - 1. Activity of proteases, strongly related with erythrocyte membrane, was analysed employing a new methodological approach. 2. Intact bovine ghosts, ghosts depleted in peripheric proteins or purified Triton X-100 and ghost extracts were electrophoresed and the proteolytic activity in the gel fragments (SDS-PAGE) was assayed. 3. At least two proteases that were inhibited by EDTA and PMSF were found. PMID- 3049184 TI - Stopped flow kinetic studies of adenosine triphosphate phosphoribosyl transferase, the first enzyme in the histidine biosynthesis of Escherichia coli. AB - 1. Stopped flow kinetic studies of ATP phosphoribosyl transferase (EC 2.4.2.17) from Escherichia coli showed that high protein concentration and elevated temperature do not give a drastic reduction of the transferase activity when measured before inhibitory amounts of PRibATP (product) has accumulated. 2. A small and slow increase in activity follows a reduction of the protein concentration showing a slow dissociation of the enzyme from an inactive to an active species. 3. By lowering the concentration of the inhibitor histidine, a fairly slow increase in activity is observed indicating a dissociation of the enzyme. 4. AMP and histidine together give a strong inhibition of the activity, while AMP alone stimulates the enzyme activity. PMID- 3049185 TI - Effect of doxycycline on pre-menstrual syndrome: a double-blind randomized clinical trial. AB - Thirty patients with well-defined symptoms of pre-menstrual syndrome were randomly treated with the antibiotic doxycycline or placebo. The antibiotic treated group showed a highly significant reduction of symptoms. Subsequent antibiotic treatment of the original placebo group similarly diminished the symptoms in this group. A 6-month follow-up demonstrated that the improvement in symptom scores was permanent and independent from the presence of the antibiotic. Luteal phase endometrial biopsies showed a high incidence of out-of-phase endometrium. An unexpectedly high percentage of endometrial biopsy cultures yielded positive findings for mycoplasma, Chlamydia trachomatis and anaerobic bacteria. There were no characteristic hormonal changes in this study group. An infectious aetiology, possibly a sub-clinical endometrial or ovarian infection, behind certain cases of pre-menstrual syndrome is postulated. PMID- 3049186 TI - Motor training and physical fitness: possible short- and long-term influences on the development of individual differences in behavior. AB - Individual differences in the behavior of young and adult animals have been documented in diverse species. Possible sources of such variation are of interest to scientists representing many disciplines, including behavior, genetics, and population and evolutionary biology. Two variables that may be important in the ontogeny and maintenance of behavioral differences are (1) individual physical (aerobic and anaerobic) fitness and (2) possible genetic variations underlying individual abilities to engage in, and to benefit from, motor training early in life. The differential development of aerobic and anaerobic capacities may play a significant role in the ontogeny of individual differences in the performance of various motor skills. There also may be short- and long-term consequences of variations in physical fitness that influence individual abilities to perform energy demanding acts during aggressive encounters, interactions with prey or predators, and courtship and breeding. Genetic studies of a limited number of species indicate that specific genotypes are correlated with individual variations in motor performance, even among conspecifics. Multidisciplinary research concerning possible relationships among the ontogeny of physical fitness, genetics, and variations in behavior is needed. Recent work on the relationship between individual differences in physical fitness and variations in the behavior of adult cold-blooded vertebrates provides a good model for comparative research on warm-blooded species. PMID- 3049187 TI - Clinical approach to the management of intractable epilepsy. AB - Resistance of seizures to anti-epileptic therapy may be due to patient error, or to physician diagnostic and/or treatment error, rather than being truly intractable epilepsy. Increased drug dosage, irrespective of blood levels, a change of drug, or the addition of a second drug, are variously indicated in truly resistant cases. The use of more than two drugs is better avoided, and in some cases reduction of treatment may improve seizure control while lessening side-effects. Repeated assessment of patients with refractory epilepsy is important, as causative or provocative factors may remain latent for long periods. Surgical therapy probably should be used more often and earlier than it now is for epilepsies that are medically intractable. PMID- 3049189 TI - Muscle and brain disease. PMID- 3049188 TI - Differentiation and oncogenes. PMID- 3049190 TI - Status of school children's hearing aids relative to monitoring practices. AB - Data concerning hearing aid monitoring practices and hearing aid malfunction were collected for three groups of mainstreamed hearing-impaired children: 248 children receiving services from itinerant teachers of the hearing impaired, 43 who participated in a study involving extensive psychoeducational evaluation, and 10 children who attended a 6 week residential treatment program in which hearing aid function was checked at least twice each day. Comparisons of the three data sets revealed that even conscientious parental and professional monitoring practices prove inadequate. As a result, it is hypothesized that children must take active roles in the monitoring process. A program to help children develop hearing aid monitoring skills is outlined. PMID- 3049192 TI - [Interhemispheric asymmetry in the mechanisms of nonaphasic disorders of the speech functions]. PMID- 3049191 TI - [Altered states of consciousness in healthy persons (the formulation of the question and research prospects)]. PMID- 3049194 TI - [HIV infection and lesions of the oral cavity: clinical and therapeutic aspects]. PMID- 3049193 TI - [Mechanisms of cellular cooperation in the normal brain and in pathology]. PMID- 3049195 TI - [Diagnosis of deep venous thrombosis of the lower limbs]. PMID- 3049197 TI - [Collateral circulation of the portal vein: evaluation and comparative study using conventional radiography of the esophagus, digestive endoscopy and abdominal echotomography]. PMID- 3049196 TI - [The role of the bone cell in the regulation of phosphorus and calcium homeostasis]. PMID- 3049198 TI - [Pulmonary edema]. PMID- 3049199 TI - [Hypo-osmolar syndrome: relation to neoplastic pathology]. PMID- 3049200 TI - [Changes in intracellular calcium with aging]. PMID- 3049201 TI - [Paraneoplastic syndromes and the kidney]. PMID- 3049202 TI - [Trace elements: endocrine-metabolic correlations]. PMID- 3049203 TI - [Complex rhythm disorders in mitral valve prolapse]. PMID- 3049204 TI - In vitro cytocidal effect of novel lytic peptides on Plasmodium falciparum and Trypanosoma cruzi. AB - Plasmodium falciparum and Trypanosoma cruzi were killed by two novel lytic peptides (SB-37 and Shiva-1) in vitro. Human erythrocytes infected with P. falciparum, and Vero cells infected with T. cruzi, were exposed to these peptides. The result, in both cases, was a significant decrease in the level of parasite infection. Furthermore, the peptides had a marked cytocidal effect on trypomastigote stages of T. cruzi in media, whereas host eukaryotic cells were unaffected by the treatments. In view of the worldwide prevalence of these protozoan diseases and the lack of completely suitable treatments, lytic peptides may provide new and unique chemotherapeutic agents for the treatment of these infections. PMID- 3049205 TI - [Cardiac transplantation: role of echocardiography in the diagnosis of rejection]. AB - Sixteen patients surviving orthotopic cardiac transplantation were studied by M Mode and two dimensional echocardiography (ECHO) on the same day of cardiac biopsy during (n = 138) a mean follow-up of 6.2 +/- 4 months (range 1-4 months). The following parameters were measured: right ventricular end diastolic internal diameter (RVdD) left ventricular end diastolic internal diameter (LVdD), interventricular septum (IVS) and posterior wall (PW) diastolic thickness, myocardial mass (MM); LV cross sectional area (CSA) and ejection fraction (EF). LV and RV wall motion, pericardial effusion and myocardial echogenicity (brightness) were evaluated by inspection. Every ECHO was compared with the previous one for qualitative and quantitative changes. In the absence of rejection, analysis of the data during the first postoperative week showed the following results: mean EF = 51 +/- 6.8%, dilated overloaded RV (30.4 +/- 4.8 mm), various amount of pericardial effusion; FE increased significantly (55.3 +/- 4%; p less than 0.001) and RVdD decreased (26.6 +/- 5mm; p less than 0.001) after the 2nd week and remained stable thereafter, while pericardial effusion decreased or disappeared. The mean values of the remaining ECHO parameters did not very significantly during the follow-up.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3049206 TI - [Autotransfusion in heart surgery]. PMID- 3049207 TI - Imaging the direct bidirectional spread of disease between the abdomen and the female pelvis via the subperitoneal space. AB - This report expands the concept of the subperitoneal space (SS) as the potential conduit for direct spread of disease in the abdomen to include the female pelvis. The normal anatomy of the SS in the lower abdomen, the female pelvis, and its uninterrupted continuation between the abdomen and pelvis are demonstrated by several imaging modalities. Surgically proven cases of bidirectional spread of disease between the abdomen and female pelvis are reported. The unifying concept of the interrelationship formed by the SS provides an understanding of the basic concepts of the pathways of direct spread of disease and the pathogenesis of the clinical presentation of disease distant from its site of origin. PMID- 3049208 TI - Duodenal duplication in the adult: its relationship with pancreatitis. AB - Seven cases of duodenal duplication in the adult are reported. The ultrasound and computed tomographic findings are analyzed. The relationship between duodenal duplication and pancreatitis is discussed. PMID- 3049209 TI - Isolated ventral pancreatitis in an alcoholic with pancreas divisum. AB - An alcoholic with no history of clinical pancreatitis was found to have pancreas divisum and marked changes of chronic pancreatitis isolated to the ventral pancreas. Pancreas divisum has been suggested to cause recurrent pancreatitis in some patients. Gross and histologic changes of pancreatitis in only the dorsal pancreas of surgically resected specimens from patients with pancreas divisum is thought to support the concept that obstruction at the minor papilla produces dorsal pancreatitis. Alternative explanations for the occurrence of segmental pancreatitis and the possible synergistic role of ethanol and bile are reviewed. PMID- 3049211 TI - Mesenchymal hamartoma of the liver in an adult: radiologic diagnosis. AB - A case of mesenchymal hamartoma of the liver in a 30-year-old female is reported. Ultrasonography, computed tomography, and hepatic angiography demonstrated the presence of a large, mixed cystic and solid mass within the liver, suggesting the diagnosis of a mesenchymal hamartoma. At surgery, the tumor proved to be unresectable. PMID- 3049210 TI - Echo-guided percutaneous puncture: a safe and valuable therapeutic tool for amebic liver abscess. AB - To clarify the therapeutic role of echo-guided percutaneous puncture (EPP) in management of amebic liver abscess, 20 patients (24 abscesses) received metronidazole plus EPP. Fluid was aspirated through Chiba needles under real-time sonographic guidance so as to reduce cavity size to less than 3 cm. Not more than two EPPs were necessary in the majority of cases and no complication followed the procedure. This scheme resulted in a shortening of time of both hospitalization (less than or equal to 20 days) and liver lesion healing as assessed by ultrasound (less than or equal to 4 months). It is concluded that EPP is a valuable and safe therapeutic tool for hepatic amebic abscess. PMID- 3049212 TI - Ultrasound evaluation of cholelithiasis in the morbidly obese. AB - The ability to detect gallstones in the morbidly obese population has been questioned in recent literature. Utilizing state-of-the-art, real-time ultrasound equipment, 44 morbidly obese patients were examined prior to gastric exclusion surgery and concomitant cholecystectomy. The 91% sensitivity and 100% specificity of gallstone detection in this series matches the results for the general population. This study provided the unique opportunity to evaluate a large number (34) of negative ultrasound examinations, with subsequent surgical confirmation yielding a negative predictive value of 97%. This confirms the continued role of ultrasound in the evaluation of cholelithiasis. PMID- 3049213 TI - Nitrofurans in the treatment of gastritis associated with Campylobacter pylori. The Gastrointestinal Physiology Working Group of Cayetano Heredia and The Johns Hopkins Universities. AB - We conducted a double-blind, placebo-controlled, randomized treatment study in Peruvian adults with antral gastritis associated with Campylobacter pylori. Patients received either 400 mg of furazolidone (n = 14), 400 mg of nitrofurantoin (n = 24), or a placebo (n = 31) for 14 days. Endoscopy was carried out at baseline, 1 day after ceasing therapy, and 6 wk after the end of treatment to verify colonization by C. pylori and determine the extent of gastric inflammation. Treatment with nitrofurantoin or furazolidone markedly reduced or, in some cases, cleared C. pylori from the antrum (p less than 0.0005 compared with placebo). Resolution of acute gastric inflammation, as evidenced by decreased polymorphonuclear leukocyte infiltration and regeneration of the mucus layer, paralleled the reduction in bacterial colonization (p less than 0.005 compared with placebo). A high percentage of patients experienced relapse (recolonization by C. pylori and return to pretreatment levels of gastritis) within 6 wk after cessation of treatment. Significant relief of dyspeptic symptoms was not evident during the study. PMID- 3049214 TI - Duplex ultrasound measurement of postprandial intestinal blood flow: effect of meal composition. AB - Duplex ultrasound was used to evaluate the effects of 350-cal, 300-ml protein, fat, carbohydrate, and mixed (Ensure-Plus) liquid meals on celiac, superior mesenteric, and femoral artery blood flow in 7 healthy volunteers. Ingestion of separate water and mannitol solutions served as controls for volume and osmolarity. Duplex parameters of peak systolic velocity, end-diastolic velocity, mean velocity, and volume flow were determined before, and serially for 90 min after, ingestion of each test meal. Maximal changes were compared with baseline values. There were no significant changes in any of the blood flow parameters derived from the celiac or femoral arteries after any test meal ingested. In contrast, maximal changes in all superior mesenteric artery parameters were increased significantly over baseline (p less than 0.05) after each of the test meals except water, with end-diastolic velocity showing proportionally the greatest increase. The study demonstrates that duplex ultrasound can provide a noninvasive means of studying the reactivity of the splanchnic arterial circulation to different stimuli and documents differing blood flow responses to variation of nutrients. PMID- 3049215 TI - Aberrant expression of monoclonal antibody-defined colonic mucosal antigens in inflammatory bowel disease. AB - Human proximal colon from patients with inflammatory bowel disease and from controls was studied by two techniques to detect tumor-associated antigen expression. A panel of four murine monoclonal antibodies that recognize tumor associated antigens was used to test purified colonic mucins for epitope expression by radioimmunoassay and to test formalin-fixed, deparaffinized sections of colon by the immunoperoxidase technique. The panel included monoclonal antibodies 19-9, B72.3, DU-PAN-2, and CSLEX1. Colonic mucins were purified from uninvolved surgical specimens by gel filtration with Sepharose 4B and cesium chloride-guanidine hydrochloride density gradient ultracentrifugation. Purified mucins from uninvolved colonic mucosal specimens from 4 of 7 patients with ulcerative colitis expressed one or more of these epitopes by radioimmunoassay, whereas mucins from 6 disease controls did not. Reactivity patterns were heterogeneous. Immunoperoxidase testing demonstrated staining with two or more antibodies in 14 of 18 involved inflammatory bowel disease segments, whereas control sections rarely stained with these antibodies, with the exception of 19-9. Sections of uninvolved mucosa from 4 of 9 patients with ulcerative colitis stained with two or more antibodies. Staining patterns were heterogeneous. The results demonstrate that colonic expression of tumor associated epitopes occurs frequently in involved segments from both patients with ulcerative colitis and with Crohn's disease, whereas only patients with ulcerative colitis frequently expressed these epitopes in uninvolved segments. PMID- 3049216 TI - Randomized, controlled trial of recombinant human alpha-interferon in patients with chronic hepatitis B. AB - Forty-five patients with chronic hepatitis B were entered into a randomized controlled trial of recombinant human alpha-interferon therapy. All patients had hepatitis B surface antigen in serum for at least 1 yr and had stable serum levels of both hepatitis B virus deoxyribonucleic acid and hepatitis B e antigen. During the 4-mo period of therapy, 10 of 31 (32%) treated patients and only 1 of 14 (7%) control patients became negative for serum hepatitis B virus deoxyribonucleic acid and deoxyribonucleic acid polymerase. All 10 patients who became negative for serum hepatitis B virus deoxyribonucleic acid subsequently had a marked improvement in serum aminotransferase activities and lost hepatitis B e antigen from serum, and 9 of them had improvement in liver histology. Comparison of responders to nonresponders indicated that female sex and a high initial level of serum aspartate aminotransferase correlated best with response to interferon therapy. These findings indicate that a 4-mo course of recombinant alpha-interferon can induce a remission in disease in approximately one-third of patients with chronic hepatitis B. PMID- 3049217 TI - Antibodies to liver/kidney microsome1 in chronic active hepatitis recognize specific forms of hepatic cytochrome P-450. AB - Anti-liver/kidney microsome1-positive sera from children with chronic active hepatitis were studied in an effort to identify the microsomal antigens selected during induction and progression of this autoimmune disease. Immunoblot analysis of sodium dodecyl sulfate gel-resolved microsomal proteins from human and rat liver using anti-liver/kidney microsome1-positive sera revealed a single polypeptide of 48 kilodaltons (human microsomes) or 50 kilodaltons (rat microsomes). Levels of the 50-kilodalton rat microsomal polypeptide were suppressed in vivo by several drugs known to modulate expression of individual forms (enzymes) of hepatic cytochrome P-450, with the largest decrease effected by phenobarbital. Dot blot analysis using a panel of 10 electrophoretically homogeneous rat liver cytochrome P-450 forms under nondenaturing conditions established that the two methylcholanthrene-inducible forms, P-450 BNF-B and P 450 ISF-G (P-450 gene subfamily IA), are selectively recognized by the anti liver/kidney microsome1 antibodies. These findings demonstrate that sera associated with autoimmune (anti-liver/kidney microsome1) chronic active hepatitis are specifically reactive with select rat hepatic P-450 forms and suggest that these autoantibodies may be principally directed against one or more constitutive forms of the corresponding human liver cytochromes. PMID- 3049218 TI - Renal prostaglandin E2 and other vasoactive modulators in refractory hepatic ascites: response to peritoneovenous shunting. AB - To assess the role of renal prostaglandin E2 in the pathogenesis of refractory ascites, in relation to renal sodium handling and circulating levels of vasoconstrictive substances, we studied 12 cirrhotic patients with refractory ascites before and after peritoneovenous shunting. Baseline values for urinary prostaglandin E2 excretion, sodium excretion, and creatinine clearance, as well as serum aldosterone, plasma renin activity, and plasma free norepinephrine, were obtained preoperatively with patients on a sodium- and fluid-restricted diet. Diuretics were also withheld. Similar parameters were measured immediately postoperatively during four consecutive 2-h intervals, then again at 2 wk and 3 mo. In patients with refractory ascites, mean baseline urinary prostaglandin E2 excretion was significantly elevated (2.5 +/- 0.8 pmol/min), compared with that in both normal controls and cirrhotics without ascites (1.3 +/- 0.3 pmol/min). A significant natriuresis occurred immediately postoperatively and persisted at 2 wk and 3 mo. Concomitantly, the elevated levels of preoperative vasoconstrictor substances gradually normalized by 2 wk. Urinary prostaglandin E2 excretion, however, rose transiently in the immediate postoperative period and then fell gradually to within the normal range by 3 mo. Enhanced renal prostaglandin E2 synthesis, therefore, does not play a role in the sustained improvement in sodium homeostasis after peritoneovenous shunting in patients with refractory ascites. PMID- 3049220 TI - Optimization of ascitic fluid culture technique. AB - The conventional method of ascitic fluid culture detects bacteria in only 42%-65% of patients who have neutrocytic ascites and suspected spontaneous bacterial peritonitis. In this study ascitic fluid was cultured by the conventional method as well as by a new method consisting of bedside inoculation of blood culture bottles with ascites. The conventional cultures grew bacteria in only 13 (43%) of 30 episodes of neutrocytic ascites, whereas the blood culture bottles grew bacteria in 28 (93%); this difference was significant (p less than 0.0001). The blood culture bottle method also resulted in more rapid detection of bacterial growth. The median concentration of bacteria in infected ascites was one organism per milliliter. Bedside inoculation of blood culture bottles with ascitic fluid is more sensitive than the conventional method in detecting bacterial peritonitis. The insensitivity of the conventional method is probably due to the low concentration of bacteria in infected ascites and the small volume of ascites cultured by this method. PMID- 3049219 TI - Sensitivity and specificity of microscopic examination of gallbladder bile for gallstone recognition and identification. AB - During cholecystectomy, gallbladder bile and gallstones were obtained from 77 patients and gallbladder bile was obtained from 39 patients free of stones (11 patients had biliary stenosis). According to their chemical composition, gallstones were classified as cholesterol (n = 46) or pigment (n = 31) stones. In patients with gallstones (a) cholesterol crystals better helped to identify cholesterol gallstones (sensitivity, 87%; specificity, 97%; positive predictive value, 97%) than did an abnormal cholesterol saturation index of bile (sensitivity, 93%; specificity, 48%; positive predictive value, 73%); (b) the presence of cholesterol crystals was significantly related to the cholesterol content of gallstones and the bile cholesterol saturation index; and (c) bilirubinate crystals, when present alone (without cholesterol crystals), were good predictors of pigment gallstones (sensitivity, 71%; specificity, 93%; positive predictive value, 88%). In the absence of stones, bilirubinate crystals were present in 9 of 28 patients without biliary stenosis (4 with alcoholic cirrhosis and 2 with alcoholic pancreatitis) and 8 of 11 patients with biliary stenosis. In the absence of stones, cholesterol crystals were present in 2 of 28 patients without biliary stenosis and in 4 of 11 patients with biliary stenosis, suggesting that bile stasis can induce cholesterol crystal formation. PMID- 3049222 TI - [The 70th anniversary of Soviet public health. The formation of the Soviet blood service]. PMID- 3049223 TI - [Effect of various factors on the destruction of abnormal hemoglobins in vitro. A screening method for the detection of unstable abnormal human hemoglobins]. PMID- 3049221 TI - Sphincter of Oddi dysfunction: a clinical controversy. AB - This report analyzes the literature on sphincter of Oddi dysfunction as it applies to biliary-type pain. The sensitivities and specificities of the tests used to diagnose this condition (e.g., size of bile duct, drainage time of bile duct, provocative tests with morphine, sphincter of Oddi manometry) are poorly defined. Recent studies suggest that noninvasive tests such as quantitative nuclear scintigraphy and fatty meal sonography may aid in diagnosing functional common bile duct obstruction. Continuous manometry of the biliary tree with microtransducer technologies may allow a greater understanding of the causes of pain in this group of patients. Only 1 case report of pharmacologic management for this disorder exists in the literature. Endoscopic sphincterotomy may be helpful in relieving the pain that occurs in this condition but is associated with increased risks. There is no consensus in the literature as to the best test that will predict response to sphincterotomy. Controlled trials of medical therapies are needed. PMID- 3049224 TI - [Tolerance for bone marrow procurement in autologous donors studied by myelokaryocytapheresis]. PMID- 3049225 TI - Heritability estimate for refractive errors--a population-based sample of adult twins. AB - The population-based Finnish Twin Cohort study was used to establish a heritability estimate for refractive errors especially for myopia. The twin cohort was derived from adult same-sexed twins in Finland. The total number of twins with both members alive in 1984 was 23,570. Of these, 3,676 twin pairs were monozygotic, and 8,109 pairs dizygotic. The sample for the present study was linked from the Finnish Police Force data base in 1984, where information of a person's possession of a driver's license and the obligation to wear glasses for far correction when driving a motor vehicle is recorded. Correlations in liability were estimated according to a multifactorial method of Smith. Falconer's heritability was 0.62 among males and 0.98 among females in the age group 28-29 years. When compared to previous twin studies of myopia, the proband concordance rates were higher for both MZ and DZ twin pairs. PMID- 3049226 TI - Sewall Wright, 1889-1988: in memoriam. PMID- 3049227 TI - Some effects of selection strategies on linkage analysis. AB - A study was designed to address the relative merits of different sampling strategies for detecting linkage. Genotypes of pedigree members were generated by the use of a single genetic model, and the pedigrees were subdivided into dominant-appearing, recessive-appearing, and "interesting" subsets. An investigator blind to how the data had been generated applied two different selection rules to determine which individuals in each pedigree would be "typed" for linkage analysis. Linkage analyses were then conducted on these pedigree subsets, as well as on the combined data, by the use of three autosomal dominant models, three autosomal recessive models, and the generating (i.e., "true") model. Results suggest (1) that linkage is likely to be detected even in the absence of knowledge of the mode of transmission, if a range of models can be examined; (2) that false evidence for heterogeneity will not necessarily result when pedigrees are selected according to apparent mode of transmission for analysis; (3) that recessive-appearing pedigrees (i.e., those with multiplex sibships) may be particularly useful for detecting linkage; and (4) that including information on unaffected second-degree relatives adds little to linkage studies of affected individuals and their first-degree relatives. PMID- 3049228 TI - A cognitive-educational treatment for hypochondriasis. AB - Hypochondriasis may be conceptualized as a disorder of perception and cognition, in which somatic sensation is experienced as abnormally intense and is then incorrectly attributed to serious medical disease. We describe a therapy to modulate the hypochondriacal patient's somatic sensations, and to alter his or her cognitive appraisal of them, which focuses on four factors that amplify somatic symptoms: (1) attention and expectation; (2) symptom attribution and appraisal; (3) the context used for interpreting the symptoms; and (4) disturbing affect and dependency needs. This therapy, or selected portions of it, can be employed in clinical work with patients individually and in groups, in consultation work, and in more traditional psychotherapeutic settings. PMID- 3049229 TI - Research links between psychiatry and cardiology. Hypertension, type A behavior, sudden death, and the physiology of emotional arousal. AB - Psychiatrists and cardiologists have collaborated in clinical care and research for decades. This article reviews some of the current areas of mutual interest, with special attention focused on research on behavior and hypertension, type A behavior pattern, sudden death, and the physiology of emotional arousal. In clinical care and epidemiologic and physiologic research, there are numerous issues at the cutting edge of knowledge that will be clarified by continued collaboration. PMID- 3049231 TI - Psychiatric services in general hospitals. Rational and irrational considerations. AB - The structure of a psychiatric service in an urban general hospital is complex. Varied intrainstitutional and extrainstitutional relationships create stress, which can lead to rational and irrational reactions, often in combination. Psychologic mechanisms that exist in individuals also occur as shared defense mechanisms in an institutional setting, serving to reduce anxiety and stress but often at the expense of accurate reality perception. Good communication can play a vital role in reducing reality distortions but is itself often blocked or impaired by the same defense mechanisms that led to the distortions. An awareness of how these mechanisms operate in an institutional setting can aid the psychiatric administrator in correcting distortions and maintaining good channels of communication. PMID- 3049230 TI - Evolution of consultation-liaison services in bone marrow transplantation. AB - Bone marrow transplantation offers new hope for cure in a variety of leukemias, aplastic anemia, and other immunodeficiency and hematologic disorders. But the psychologic circumstances accompanying this procedure are often profoundly stressful. The evolution of consultation--liaison services on a bone marrow transplantation unit are described and illustrated with several clinical vignettes. The results of a survey of 52 other bone marrow transplantation centers report the percentage that provide such services and the extent to which they are performed by a psychiatrist, a nonphysician psychosocial counselor, or both. PMID- 3049232 TI - [The relationship of cytoplasmic and mitochondrial protein synthesis in yeasts: the mapping of mitochondrial mutations neutralizing the exhibition of nuclear omnipotent suppressors]. AB - The genetic and physical mapping of mitochondrial mutations [CRD] neutralizing respiratory deficiency in sup1 and sup2 mutants was performed. The genetical methods demonstrated improbability of location of these mutations in genes coding for the enzymes of respiratory chain and for 21S rRNA. Southern-blot analysis has shown these mutations to be localized in the Hinc10 fragment of mitochondrial genome. This fact was interpreted as indication that var1 gene is affected by [CRD] mutations. Our results are in agreement with the hypothesis on the participation of sup1(2) proteins in mitochondrial translation. PMID- 3049236 TI - [Genetic control of plasmid recombination in the cotransformation of Saccharomyces cerevisiae: the role of RAD52 and FLP genes]. AB - In our earlier works we observed high frequency of recombination between two chimeric plasmids of different types, when they were introduced into yeast cells via cotransformation. Incapability of one of these plasmids to replicate autonomously in yeast cell is the necessary condition for such recombination. The high efficiency of this process point to the differences between interplasmid recombination and other types of yeast recombination. In this work, we studied the participation of two genes in the control of interplasmid exchanges. These are RAD52 responsible for normal processes of meiotic and mitotic recombination and highly specific gene FLP located on 2 mkm DNA which specifies site-specific recombination in the region of inverted sequences of this plasmid. The mutation rad52 in the recipient strain was shown to sharply decrease the efficiency of recombination between integrative and episome plasmids during cotransformation. The absence of FLP gene in the recipient strain (cirO) has no influence on this process. PMID- 3049233 TI - [Macromutation and evolution: the fixation of Goldschmidt's macromutations as species and genus traits. Hairlessness mutations in mammals]. AB - A brief survey of the development of concepts on the role of macromutations in evolution is given. Contrary to Iu. A. Filipchenko (1926, 1927), who introduced the "micro- and macromutation" terms and believed that regularities of macroevolution could not be reduced to microevolutionary processes, the majority of "synthetists" explained any form of evolution by changes in allele frequencies. From the studies of Drosophila homoeotic mutants R. Goldschmidt (1940) developed the concept of "hopeful monsters" and their role in macroevolution. However, the homoeotic mutants are of drastically reduced viability, which allows the gradualists to reject Goldschmidt's ideas. The distribution of hairlessness mutations (hairless, nude etc.) with the monogenic pattern of inheritance in mammals was studied. Hairless mutants are known in Peromyscus, Mus musculus, Rattus rattus, R. norvegicus, Canis familiaris, Ovis aries. Hairlessness as norm is found in 53 among contemporaneous 1037 mammalian genera. Part of these cases (hairlessness in all Cetacea and Sirenia) may be explained in terms of both macromutations and obligatory gradualism. There is no doubt as to the macromutational origin of hairlessness in the bat Cheiromeles and the rodent Heterocephalus (Bathyergidae); the genera systematically and ecologically close to these have normal pelage. It is quite possible that hairlessness of walrus (Odobenus) has the same origin. The appearance and fixation of single Goldschmidt's macromutation cannot yet be considered as a macroevolutionary process, though the possibility of fixation of a macromutation in nature as a species and genus character contradicts strongly the concept of obligatory gradualism of evolution. PMID- 3049234 TI - [The genetics of breast cancer, population and familial research and segregation analysis]. AB - The data on clinico-genealogy studies of 1046 probands with breast cancer and their relatives are presented. The nature of inheritance corresponded to the Mendelian model. As to other families, there is no strong evidence for the monogene model both with complete and incomplete penetrance of mutant homo- and heterozygotes. Penetrance of homozygotes was 7.9-30.5%, this being 2.0-7.3% for heterozygotes. The conclusion is drawn that it is necessary to consider the regularities of inheritance of breast cancer in the light of the multifactorial model. PMID- 3049235 TI - [Repetitive sequences of the human genome]. AB - A review of literature data on different families of repetitive sequences of human genome is presented. Both tandemly organized (classical satellite DNA, alpha satellite DNA, "minisatellites" etc.) and interspersed repetitive elements are characterized in detail. Special attention is paid to description of human genome mobile elements. PMID- 3049238 TI - Genomic imprinting. PMID- 3049239 TI - Phase variation in Salmonella: analysis of Hin recombinase and hix recombination site interaction in vivo. AB - The bacteriophage P22-based challenge phase selection was used to characterize the binding of Salmonella Hin recombinase to the wild-type hixL and hixR recombination sites, as well as to mutant and synthetic hix sequences in vivo. Hin recombinase binds to the hixL or hixR recombination sites and represses transcription from an upstream promoter in the challenge phage system. Hin mediated repression results from Hin associating into multimers either prior to binding or during the binding process at the hix operator sites (cooperativity). The ability of Hin multimers to repress transcription is eliminated when the hix 13-bp half-sites are rotated to opposite sides of the DNA helix by inserting 4 bp between them. Insertion of 1 bp between half-sites reduces overall repression. Hin also binds one of the hixL half-sites to repress transcription, but only when high levels of Hin protein are present in the cell. Mutations have been identified in the hix sites that impair Hin binding. Five of the 26 bp in the hix sites are critical; sites with base-pair substitutions at these five positions show greatly reduced binding. Three additional base pairs make minor contributions to binding. These results are consistent with the results of binding studies between Hin and the hix sites in vitro. PMID- 3049237 TI - Expression, modification, and localization of the fushi tarazu protein in Drosophila embryos. AB - The fushi tarazu (ftz) protein of Drosophila is required during embryogenesis for the process of body segmentation. To study the biochemical properties of the ftz protein, ftz cDNA was expressed in Escherichia coli and the protein was purified to homogeneity. Polyclonal antibodies raised against the purified protein were used to localize and quantitate the protein during embryogenesis. Three temporally and spatially distinct phases of expression were observed, which include a previously undetected period later in embryogenesis. During this last phase, the protein is localized predominantly in the developing hindgut. Analysis of embryonic ftz protein on Western blots permitted us to approximate the number of protein molecules per nucleus. During the blastoderm phase of development, when ftz protein is most abundant, we estimate that there are approximately 20,000 molecules of protein per ftz-expressing nucleus. The embryonic ftz protein migrates more slowly on SDS-polyacrylamide gels than protein made either in E. coli or in a reticulocyte lysate system in vitro, indicating that it is modified in the embryo. To facilitate characterization of ftz protein made in embryos, an ftz overexpression system functional in Drosophila was developed. When fused to an hsp70 heat shock promoter and introduced into the germ line by P-element mediated transformation, ftz could be overexpressed at all stages of development by heat shock. This protein is localized in the nucleus comigrates on SDS polyacrylamide gels with endogenous ftz protein. Two-dimensional gel electrophoresis followed by Western blotting resolves the overexpressed protein into a series of isoforms that differ in charge and electrophoretic mobility. Post-translational modification may influence the biochemical properties and functions of the ftz protein during embryogenesis. PMID- 3049240 TI - Searching for the HIV (human immunodeficiency virus) in south Sudan, Guinea Bissau, Cape Verde Islands, Iran, Nicaragua, El Salvador, Columbia. AB - In the spring of 1986, 3.473 human blood samples were serologically screened for HIV-antibodies. The methods used were ELISA (enzyme-linked immunosorbent assay), immunoblotting (Western Blot) and the indirect immunofluorescence assay (IFA). The blood samples were collected from males and females of all age groups in: South Sudan (Melut District in 1981 and 1983), Guinea Bissau (1983), on the Cape Verde Islands (1983/84), in Iran (1985), Nicaragua (1984), El Salvador (1984) and Columbia (1984). 18 out of 1.614 sera from South Sudan, 5 out of 93 sera from Guinea Bissau, 1 out of 289 tested sera from El Salvador were confirmed to be positive. None of the sera from Iran and Nicaragua were HIV-antibody positive. PMID- 3049242 TI - Serological investigations for antibodies against Treponema pallidum on the Cape Verde Islands (Santiago and Santo Antao)--2. report. AB - On the Cape Verde Islands, Santiago and Santo Antao 581 human blood samples were collected. The VDRL and TPHA tests were employed in the serological examinations of samples for antibodies against Treponema pallidum. Positive reactions were shown in 2 of the 191 sera from the Santiago Island and in 12 of the 390 sera from the Santo Antao Island in both the VDRL and the TPHA tests. Including the alone TPHA-positives of 5 and 8 sera respectively, results in a 3.7% of positive sera for Santiago and a 5.1% for Santo Antao. By not taking into account children under 13 years of age, the positivity rises to 4.2% and 7.6% respectively. Giving a total contamination rate of 6.3% for Treponema pallidum. PMID- 3049241 TI - On the drinking water situation on the Cape Verde Islands (Islands of Santiago). AB - On the Cape Verde Islands drinking water investigations were made. The drinking water was tested in chemical and bacteriological respects as well as on amoebas and rotaviruses. The isolated germs were investigated on their reaction against several antibiotics and chemotherapeutics by means of the agar diffusion test. The changing of drinking water quality from water supply places to households was investigated as well. The results make evident the urgency of extensive sanitations of water supply places on the Cape Verde Islands. PMID- 3049243 TI - Albert Szent-Gyorgyi in Szeged. PMID- 3049245 TI - Transcript characterisation, gene disruption and nucleotide sequence of the Saccharomyces cerevisiae WH12 gene. AB - WH12 is a gene which plays a prominent role in regulating growth and proliferation in Saccharomyces cerevisiae. It is expressed as a 2.0-kb mRNA transcript. Disruption of this transcript in a WH12+ cell results in the mutant phenotype being displayed. The nucleotide sequence of the cloned gene has been determined and found to include a 487-codon long open reading frame coding for a 55.3-kDa protein. The protein showed no extensive homologies to any previously identified protein. The 5' and 3' noncoding regions contained many of the features found in other yeast genes. PMID- 3049244 TI - High-level expression of a functional human fibrinogen gamma chain in Escherichia coli. AB - Human fibrinogen gamma chain has been expressed intact at high levels in Escherichia coli. The construction of the expression plasmid, p253, is described. Synthesis of the recombinant protein is isopropyl-beta-D-thiogalactopyranoside dependent and is driven by the tac promoter. Western analysis of E. coli lysates demonstrates a novel protein of approx. 45 kDa which cross-reacts with antisera to human fibrinogen gamma chains. The protein is not soluble in common E. coli lysis buffers and becomes soluble in 6 M guanidine.HCl or 6 M urea. Initial insolubility is due to interchain disulfide bond formation and to noncovalent interactions. Induced cells examined by phase-contrast microscopy contain dense inclusion bodies. A known function of the gamma chains of human fibrinogen is the clumping of Staphylococcus aureus Newman D2C cells [Hawiger et al., Biochemistry (1982) 1407-1413]. We demonstrate that suspensions of recombinant gamma chains retain the ability to clump cells from this strain of S. aureus. PMID- 3049246 TI - Expression, purification and characterization of recombinant human insulin-like growth factor I in yeast. AB - Insulin-like growth factor I (IGF-I) is a 70 amino acid (aa) protein that is structurally similar and functionally related to insulin. We have inserted a synthetic gene coding for human IGF-I into a Saccharomyces cerevisiae expression vector utilizing the MF alpha 1 promoter and pre-pro leader peptide. This vector directs the expression and secretion of native, biologically active growth factor. Cleavage of the pre-pro alpha factor leader sequence in vivo results in the secretion of a 70-aa recombinant IGF-I molecule with the native N-terminal glycine residue. Human IGF-I purified from yeast culture supernatant is equipotent to serum-derived IGF-I in inhibiting [125I]IGF-I binding to type-I IGF receptors and crude human serum-binding proteins. Recombinant IGF-I is also equipotent to human IGF-I in the stimulation of DNA synthesis in rat aortic smooth-muscle cells. In contrast, yeast recombinant IGF-I is less potent than serum-derived IGF-I in binding to type-2 IGF receptors. The ability to produce native, biologically active IGF-I in yeast will allow the elucidation of binding domains through the expression and characterization of specific structural analogs. PMID- 3049247 TI - The nucleotide sequence of a plasmid determinant for resistance to tellurium anions. AB - The plasmid pMJ606 contains a 5-kb insert specifying resistance to tellurium salts (TeR) which was cloned from the large conjugative plasmid pMER610. Nucleotide sequence analysis of this insert has identified five open reading frames (ORFs). ORFs 1, 2, 4 and 5 correspond in terms of their predicted polypeptide products to the 41, 15.5, 22 and 23-kDa polypeptides, respectively, which are synthesised by the resistance determinant in maxicells. An additional ORF, ORF3, whose product has not been identified is predicted by the sequence data. The sequence of the presumptive polypeptide specified by ORF3 indicates a membrane location. The nucleotide and predicted amino acid sequences of ORF4 and ORF5 show considerable homology which is consistent with the duplication and minor divergence of an ancestral gene. ORFs 4 and 5 appear to retain the same function and while each can function separately and contribute to the resistance mechanism, the full level of wild-type resistance requires both genes to function. The transcriptional signals for the TeR determinant may be located beyond and 5' of the sequences cloned in pMJ606. PMID- 3049248 TI - Nucleotide sequence and transcriptional analysis of a third function (Flm) involved in F-plasmid maintenance. AB - The leading region of the conjugative F plasmid encodes for a function, Flm, capable of extending the maintenance of normally unstable plasmids. Nucleotide sequencing and functional studies of flm locus have shown that it consists of at least two genes, flmA and flmB, which are physically and functionally homologous to hok and sok of parB in plasmid R1. The 52-amino acid flmA-coded polypeptide is almost identical to the hok product which has been shown to be a membrane associated lethal protein [Gerdes et al., EMBO J. 5 (1986) 2023-2029]. Gene flmB codes for a 100 nucleotide, non-translated, complementary RNA which overlaps the 5' leader sequence of the flmA RNA. The flmA RNA also encodes an open reading frame (ORF70) which overlaps the flmA-coding sequence and may be a third gene involved in the Flm function. S1 analysis and functional studies suggest that the antisense flmB RNA binds to the flmA RNA and suppresses the expression of the lethal product, presumably by blocking coupled translation of ORF70 and flmA. Secondary structure analysis predicts that the flmA RNA is extremely stable compared to the regulatory flmB RNA. We suggest that when these RNA species are retained by cells which have lost the F plasmid, the more stable flmA RNA will eventually be translated thus leading to cell death. This phenomenon provides a third mechanism, additional to ParFIA and Ccd functions, to ensure maintenance of the F plasmid in a growing bacterial population. PMID- 3049252 TI - Construction of versatile Escherichia coli--yeast shuttle vectors for promoter testing in Saccharomyces cerevisiae. AB - The promoterless PHO5 gene of the yeast Saccharomyces cerevisiae, encoding the repressible acid phosphatase (AP) was utilized as a reporter gene for the construction of a novel vector system for selection and functional analysis of yeast promoters. The Escherichia coli-yeast shuttle plasmids, pZHB81 and pZHB82, contain different arrays of unique restriction sites located upstream of the PHO5 coding region. Yeast promoters could be screened from random DNA fragments (cloned in the upstream sites) for their ability to direct the expression of the PHO5 gene in transformed (AP-deficient) yeast host cells. AP-expressing transformants were selected directly on agar plates by using a routine colony staining method. Relative promoter strength was assessed by direct assay for AP activity in cell lysates. PMID- 3049251 TI - Studies on the structure, expression and function of the yeast regulatory gene PHO2. AB - The yeast regulatory gene PHO2 encodes a protein assumed to activate, in concert with the PHO4 protein, the transcription of the repressible acid phosphatase coding gene PHO5. The PHO2 gene was cloned and sequenced. Northern-blot analysis revealed a low and Pi-independent transcription level. We stress the presence of two potential DNA-binding structures, one of them as part of a homeodomain similar sequence, in the deduced PHO2 protein. Our data indicate that quite a large portion of the C-terminal end of the PHO2 activator is dispensable for derepression of PHO5. Disruption analysis suggests the presence of a nuclear address signal in the N-terminal region. We show that the PHO2 function in the regulation of phosphate metabolism can be partially fulfilled by overproduced PHO4. Inability of pho2 mutants to sporulate indicates a possible involvement of PHO2 products in the regulation of genes related to the life cycle. PMID- 3049250 TI - Expression and excretion of human pancreatic secretory trypsin inhibitor in lipoprotein-deletion mutant of Escherichia coli. AB - We constructed a gene coding for the 56-amino acid human pancreatic secretory trypsin inhibitor (PSTI), and ligated it on a plasmid downstream from the trp promoter and the signal peptide sequence of alkaline phosphatase. The resulting plasmid was transfected into a lipoprotein deletion mutant (Escherichia coli JE5505) and the plasmid-carrying cells were induced with 3-indoleacrylic acid. A considerable amount (50 micrograms/ml culture) of the mature PSTI protein was detected in the culture supernatant. The excreted PSTI was identical to the natural PSTI protein with respect to the trypsin-inhibiting activity, the N terminal and the C-terminal amino acid sequences and the amino acid composition. PMID- 3049249 TI - Highly efficient positive selection of recombinant plasmids using a novel rglB based Escherichia coli K-12 vector system. AB - We have developed pBR328-derived vectors which allow highly efficient positive selection of recombinant plasmids. The system is based on the rglB-coded restriction activity of Escherichia coli K-12 directed against 5-methylcytosine (5mC)-containing DNA. The vectors code for cytosine-specific, temperature sensitive DNA methyltransferases (ts-Mtases), whose specificity elicits RglB restriction. 5mC-free vector DNA - a prerequisite to allow establishment of such plasmids in cells expressing the RglB nuclease activity - can be prepared from cultures grown at 42 degrees C. At 30 degrees C the vector plasmids are vulnerable to RglB restriction due to the expression of suicidal Mtase activity. Cloning a DNA fragment into the ts-Mtase-coding gene disrupts the lethal methylation and thus permits selection of such recombinant plasmids at 30 degrees C. The standard vector used, pBN73, contains unique recognition sites for nine restriction enzymes within the ts-Mtase-coding gene, which can be used independently or in combination for the construction of recombinant plasmids selectable by the rglB-coded activity. Plasmid pBN74, which carries the determinants for both the ts-Mtase and the RglB nuclease, contains seven unique sites within the ts-Mtase-coding gene. While selection of recombinant plasmids derived from pBN73 obligatorily requires the employment of rglB+ strains, selection of pBN74 derivatives can be performed independent of the E. coli-host genotype. It remains to be elucidated whether positive selection of pBN74-derived recombinant plasmids can also be achieved in hosts other than E. coli. Plasmids pBN73, pBN74 and the recombinants are structurally stable. Generally applicable procedures, as developed during the establishment of this vector system, are described; they allow the isolation of ts-Mtases and facilitate the cloning of genes coding for nucleases directed against 5mC-containing DNA. PMID- 3049253 TI - Effects of dominant-negative lac repressor mutations on operator specificity and protein stability. AB - The specific binding of dominant-negative (I-d) lactose (lac) repressors to wild type (wt) as well as mutant (Oc) lac operators has been examined to explore the sequence-specific interaction of the lac repressor with its target. Mutant lacI genes encoding substitutions in the N-terminal 60 amino acids (aa) were cloned in a derivative of plasmid pBR322. Twelve of these lacI-d missense mutations were transferred from F'lac episomes using general genetic recombination and molecular cloning, and nine lacI missense mutations were recloned from M13-lacI phages [Mott et al., Nucl. Acids Res. 12 (1984) 4139-4152]. The mutant repressors were examined for polypeptide size and stability, for binding the inducer isopropyl beta-D-thiogalactoside (IPTG), as well as binding to wt operator. The mutant repressors' affinities for wt operator ranged from undetectable to about 1% that of wt repressor, and the mutant repressors varied in transdominance against repressor expressed from a chromosomal lacIq gene. Six of the I-d repressors were partially degraded in vivo. All repressors bound IPTG with approximately the affinity of wt repressor. Repressors having significant affinity for wt operator or with substitutions in the presumed operator recognition helix (aa 17-25) were examined in vivo for their affinities for a series of single site Oc operators. Whereas the Gly-18-, Ser-18- and Leu-18-substituted repressors showed altered specificity for position 7 of the operator [Ebright, Proc. Natl. Acad. Sci. USA 83 (1986) 303-307], the His-18 repressor did not affect specificity. This result may be related to the greater side-chain length of histidine compared to the other amino acid substitutions. PMID- 3049254 TI - The primary structure of the operons coding for Shigella dysenteriae toxin and temperature phage H30 shiga-like toxin. AB - Nucleotide(nt) sequences were determined for the toxin (SHT) operon present in the chromosome of Shigella dysenteriae 1 and for the shiga-like toxin (SLT) operon found in the lambdoid phage H30 genome. The coding sequences of the sht and slt genes differ in 4 nt with 1 nt change responsible for an amino acid replacement. The deduced amino acid sequence in the A chain of the toxins is highly homologous to that of the A chain of ricin, a plant toxin. SHT-coding mRNAs were detected by mapping the 5' termini and using blot-hybridisation; one of them was more abundant and coded only for the B subunit of SHT while the other (bi-cistronic mRNA) encoded both subunits. An IS element related to the IS3 element of Escherichia coli was found in the chromosome of S. dysenteriae near the sht operon. PMID- 3049256 TI - Secretion of Escherichia coli chloramphenicol acetyltransferase by mammalian cells. AB - We show here that expression of the Escherichia coli cat gene in mammalian cells results in accumulation of enzymatically active CAT in the culture media as well as in the cytoplasm. We call the extracellular product secreted CAT (sCAT). Three to four days after introduction of cat-expressing plasmids into mouse L cells by transient transfection, total extracellular sCAT activity exceeds total cytoplasmic CAT activity. As sCAT levels increase, substantially more CAT is found outside the cells than inside at later times. Comparison of different populations of cat-expressing cells shows that, at any given time, the level of sCAT is proportional to the level of intracellular CAT. Thus, assay of sCAT provides a convenient, non-invasive alternative to assay of intracellular CAT. The molecular sizes of sCAT and intracellular CAT are indistinguishable, suggesting that the protein is not cleaved or glycosylated during secretion. Several observations, including a lack of sensitivity to drugs which inhibit Golgi activity, suggest that CAT may be secreted via an unusual pathway. PMID- 3049255 TI - Molecular characterization of yeast regulatory gene CAT3 necessary for glucose derepression and nuclear localization of its product. AB - The yeast regulatory gene CAT3 has an essential function for the depression of several glucose-repressible enzymes. Therefore, cat3 mutants are unable to grow on maltose or on non-fermentable carbon sources. Unlike the point mutants isolated previously, cat3 null allele strains also failed to utilize raffinose or galactose as sole carbon sources. Sequencing of an 1.6-kb HindIII-BglII fragment complementing cat3 mutations revealed an open reading frame of 322 codons, size of which is in good agreement with the 1.3-kb size of mRNA. No significant similarities with previously sequenced genes could be detected. CAT3-lacZ fusions confirmed the proposed reading frame. A CAT3-lacZ fusion encoding 307 amino acids of CAT3 was able to complement the growth defects of cat3 point mutants and null allele strains. Assay of beta-galactosidase activity under different growth conditions indicated a constitutive expression of the CAT3 gene product. Cellular fractionation studies showed the nuclear localization of the CAT3 protein. PMID- 3049257 TI - A series of shuttle vectors using chloramphenicol acetyltransferase as a reporter enzyme in yeast. AB - Reports from numerous laboratories have shown that the gene coding for the bacterial enzyme chloramphenicol-3-O-acetyltransferase can be used as a reporter gene (cat) in mammalian and plant systems to analyze gene activity at the transcriptional level when combined with endogenous regulatory signals; the enzyme activity can be quantified by a chromatographic or a photometric assay. To adapt this simple and highly sensitive test for the yeast system, we constructed a series of yeast vectors containing the cat gene together with selectable markers for Escherichia coli and yeast; integrating, autonomously replicating and centromere-carrying plasmids were used. We show that the cat gene lacking the endogenous promoter is expressed at low levels in yeast transformants. To demonstrate functional expression of the cat gene placed under the control of a yeast promoter, we chose the PHO5 regulatory region. We found that cat expression was induced via the PHO5 promoter in a manner as observed for the endogenous PHO5 gene, whereas in the repressed state cat expression remained low. Using these vectors, it should be feasible to analyze other sequences conferring promoter activity or other control functions in yeast. PMID- 3049258 TI - Proteins of the extracellular matrix in vitreoretinal membranes. AB - Epiretinal and vitreous membranes of different etiology, e.g., in diabetic retinopathy, following retinal detachment, trauma or inflammatory processes, show a similar morphology. The exact composition of the extracellular matrix and the pathogenesis of these membranes remain uncertain. The presence of collagens, type I-IV, laminin, and fibronectin can be shown by means of immunofluorescence with affinity-purified antibodies. Collagen type V was revealed by SDS-polyacrylamide gel electrophoresis. These proteins of the extracellular matrix have diverse properties and functions in the membranes, as is discussed. Despite great similarities in morphology, there are some differences in the matrix, seemingly dependent upon the etiology of the membrane. PMID- 3049259 TI - Structure of the vitread face of the monkey optic disc (Macaca mulatta). SEM on frozen resin-cracked optic nerveheads supplemented by TEM and immunohistochemistry. AB - The authors applied frozen resin cracking after hexamethyldisilazane (HMDS) desiccation on the monkey optic disc region. The cracked face through the central part of the optic disc showed that the inner limiting membrane of the retina continued into the limiting membrane of Elschnig, and this, in turn, continued into the central meniscus of Kuhnt. At the disc margin the membrane was about 70 nm in thickness, due to a large fibrillar component. Elschnig's membrane was about 50 nm in thickness and was composed of both fibrils and flocculent material. The membrane covering the central meniscus of Kuhnt was about 20 nm in thickness. The number of fibrils here was very low, and the membrane consisted of flocculent material. The positive immunohistochemical stainings for GFA and vimentin of Elschnig's membrane and Kuhnt's meniscus were noteworthy. The positive staining disappeared when the membrane continued into the inner limiting membrane of the retina, supporting the different structural composition. PMID- 3049260 TI - Diffusion barriers impeding the flow of solutes to the ciliary body epithelium during immersion fixation of rabbit eyes. Assessment of ultrastructural preservation quality by examination of mitochondrial profiles in the electron microscope. AB - Various manipulations of rabbit anterior eye segments were carried out during the initial stages of glutaraldehyde fixation in order to assess the contribution made by particular tissues in restricting the flow of solutes to the ciliary body epithelium. Ultrastructural preservation quality was monitored by inspection of mitochondrial profiles. These organelles are structurally extremely sensitive to environmental disturbances, and hence changes in their morphological appearance may be used as an early indication of such adverse conditions. The results indicate that the principal barriers to diffusion are represented by the vitreous, posteriorly, and by the cornea at the anterior face. Since the cortical vitreous is strongly attached to the inner basement membrane of the ciliary epithelium, its removal may cause damage to this layer. A simpler and equally effective means of improving flow of fixative to the ciliary epithelium is to remove the cornea. PMID- 3049261 TI - Is routine second-look laparotomy for ovarian cancer justified? AB - Thirty-nine patients with epithelial ovarian malignancy underwent second-look laparotomy (2LL), as part of their plan of management at the Johannesburg University Hospital. Twenty-eight patients (71.8%) were found to have no gross or microscopic evidence of disease. Only 1/12 (8.3%) of patients with initial Stage I disease had evidence of persistent disease and after a median follow-up of 53 months (range 29-77) after 2LL, the remaining 11 remain free of relapse. Second look laparotomy is regarded as unjustified in this subgroup of patients. Twenty nine percent of the patients with advanced disease (Stage III and IV) who were disease-free at 2LL subsequently developed recurrent disease and died. In this group 2 additional patients died of nonmalignant disease. All 3 of the patients with original Stage II disease were disease-free at 2LL, but subsequent recurrence developed in 1 patient. On the basis of the findings in this study and evidence in the literature, the practice of submitting patients who are in complete clinical remission to 2LL as part of their management plan is questioned and challenged. PMID- 3049262 TI - Lymphography in the initial evaluation of endometrial carcinoma. AB - Two hundred eighty-eight patients with endometrial carcinoma underwent a bipedal lymphography. The proportion of positive lymphangiograms is related to the clinical stage, the histologic grade, and the depth of myometrial invasion. Lymphography is not a significant predictor of survival taking into account the other prognostic factors. A histological examination of the lymph nodes was carried out for 138 patients. Lymphography is not very sensitive but is highly specific, detecting only 50% of metastases with a false positive rate that is too high. It therefore is of little diagnostic and prognostic value for operable patients. It is, however, useful for the followup of lymph nodes of patients treated by radiation therapy. PMID- 3049263 TI - Successful management of malignant pericardial effusion in metastatic squamous cell carcinoma of the uterine cervix. AB - Malignant pericardial effusion secondary to pericardial metastases from gynecological malignancies represents an infrequent but potentially life threatening problem. A patient with recurrent squamous cell carcinoma of the cervix causing symptomatic pericardial effusion is presented, and the incidence, mechanism, pathophysiology, treatment, and outcomes of malignant pericardial effusion in patients with gynecologic malignancies are reviewed. This case represents only the fourth reported patient with metastatic carcinoma of the cervix in whom the diagnosis of malignant pericardial effusion was made antemortem, and is the longest survivor of treatment. Gratifying results, in terms of improved quality and length of survival, can be obtained in what is often perceived as a preterminal complication. Recommendations for management are presented, stressing radiation therapy and other local measures following initial pericardiocentesis. PMID- 3049265 TI - Ultrasonographic evaluation of adrenal involution during antenatal and neonatal periods. AB - Using real-time ultrasonography, 25 adrenal glands from prenatal to neonatal periods were prospectively examined to assess the process of shrinkage. A similar sonographic structure was seen in both prenatal and neonatal adrenal glands. The area of adrenal gland (AGA) and the rate of decrease were calculated during antenatal and early neonatal periods. The values of AGA were 350 +/- 19 mm2 within a week of delivery, 304 +/- 17 mm2 (87 +/- 2%) just after delivery, 273 +/ 25 mm2 (77 +/- 5%) on the 1st day, 246 +/- 24 mm2 (70 +/- 5%) on the 2nd day, 215 +/- 23 mm2 (61 +/- 5%) on the 3rd day, 196 +/- 22 mm2 (56 +/- 5%) on the 4th day, 173 +/- 18 mm2 (50 +/- 4%) on the 5th day, 154 +/- 14 (44 +/- 4%) on the 6th day, 140 +/- 12 mm2 (40 +/- 2%) on the 7th day, respectively. In conclusion, the postnatal involution of the adrenals was documented by ultrasound. PMID- 3049264 TI - [Tolerance and acceptance of the norethisterone containing triphasic pill Trinovum]. PMID- 3049267 TI - Comparison of transabdominal and transvaginal pelvic ultrasonography for ovarian follicle assessment in in vitro fertilisation. AB - The emergence of transvaginal ultrasound-guided oocyte retrieval provided an opportunity to review and improve the traditional transabdominal ultrasonic approach for follicular tracking in in vitro fertilisation (IVF). This technique requires a full bladder, which may cause extreme discomfort. Hence, to provide both effective and comfortable monitoring, we instituted a study comparing transabdominal and transvaginal ultrasonography, with regard to patient preference and follicular number, size and dominance, in patients undergoing IVF and gamete intrafallopian transfer (GIFT). Commencing usually on day 9 of the treatment cycle, 45 patients were scanned on 55 occasions, initially abdominally (with a full bladder) and subsequently vaginally, using a transvaginal 7.5 MHz sector transducer. Follicular number was identical in 78% of cases, with the majority of the remaining patients showing an extra follicle on vaginal assessment. There were no significant differences in overall or dominant follicular diameters with either technique. Overall, 85% unashamedly preferred the vaginal approach. We believe that tracking follicular development in IVF treatment cycles is efficient and popular using the vaginal sector transducer and accordingly have ceased all assessments using the abdominal probe. Subsequently, 450 follicle scans have been performed with virtually the unanimous approval of our patients. PMID- 3049266 TI - Suppression of puerperal lactation by terguride. A double-blind study. AB - Clinical efficacy, prolactin (PRL)-lowering effect and tolerance of terguride (an 8-alpha-ergoline derived from Lisuride which acts as a partial dopaminergic agonist) were investigated in a double-blind study on inhibition of puerperal lactation using three different daily doses of the drug (0.25, 0.5 and 1.0 mg). With 0.5 and 1.0 daily therapeutical regimens PRL levels were suppressed in a dose-dependent manner and lactation was prevented. Terguride was highly well tolerated. PMID- 3049269 TI - [The quadriga phenomenon of the extensor tendon system and the superficial transverse metacarpal ligament]. AB - The aponeurosis of the common extensor tendons is connected by intertendinous connections proximal to the metacarpophalangeal joints. These fibers are responsible for tenodesis effects in case of maximal finger extension and flexion and by this means they diminish the function of the other metacarpophalangeal joints. This mechanism can be described as the Quadriga-Syndrome of the extensor apparatus. These connecting structures also help to stabilize the transverse metacarpophalangeal arch and the position of the extensor tendons over the metacarpal heads. Cadaver dissections as well as operative findings have shown that there exists a superficial transverse intermetacarpal ligament, which connects the metacarpal heads in the area of its neck. PMID- 3049268 TI - Ultrasonic prediction of stress urinary incontinence development in surgery for severe pelvic relaxation. AB - Forty-six patients with a third-degree cystocele with no complaints of genuine stress urinary incontinence underwent complete urodynamic evaluation including ultrasonic evaluation before and 3 months postoperatively. Ultrasound examination was able to predict in 46 patients those requiring urethropexy surgery for support of their bladder base (n = 22). The 24 patients with good bladder base support by ultrasound did not develop genuine stress urinary incontinence. The use of ultrasound in women with third-degree cystocele appears to predict which patients will require urethropexy surgery in addition to their cystocele repair. PMID- 3049270 TI - [Technic and problems of resection of small intestine segments for free transplantation]. AB - The removal of a segment of the small bowel for free microvascular transplantation corresponds to a small bowel resection in routine abdominal surgery. Attention must be given to the careful preparation of a sufficiently long vascular pedicle, clear separation and marking of the artery and vein, and adaptation of the mesentery to the graft location. According to the author's experience inflammatory small bowel disease and general occlusive vascular disease are contraindications to small bowel transplantation. PMID- 3049271 TI - [What significance does superselective angiography of the hip joint have in femur head necrosis of the adult before and after management with a vascular pedicled ileal crest graft?]. AB - The use of microsurgical techniques in orthopaedics makes it possible to revascularize the necrotic femoral head in adults. Superselective angiography improves preoperative planning. Furthermore, using this method postoperative perfusion control of the pedicle bone graft can be carried out. This paper reports on a special way of positioning the pelvis, as well as on the results of angiography in various functional positions of the hip joint. PMID- 3049272 TI - [Partial intercarpal arthrodesis]. AB - An intercarpal arthrodesis was performed in 21 patients with Kienbock's disease, scaphoid non-union, and instability of the wrist. In eight patients we used the method of Graner, in eight cases the procedure was an arthrodesis between scaphoid and lunate and in five cases an arthrodesis between other carpal bones. Eighteen of these patients were reviewed one to 14 years postoperatively. 30% were pain free, 40% had some residual pain, and 20% had moderate to severe pain. The range of motion in the wrist was restricted mainly in extension. In 40% we found arthrotic changes. Two cases resulted in an arthrodesis of the wrist. Based on the poor results found in this review, we feel the indications for intercarpal arthrodesis are very limited. PMID- 3049273 TI - [What results can be expected following the biceps replacement operation? An analysis of retrospective data]. AB - The authors reviewed the literature and analysed the results of Steindler's technique, the transfer of pectoralis major muscle and its modifications, and the transfer of latissimus dorsi muscle. They determined the average extension lag, flexion, mobility, and strength in the publications they reviewed. Their own results were included in the statistics. The analysis of the results of the various techniques showed, 1. There is not always good correlation between the theoretical advantages and disadvantages of a surgical method and the clinical results of a particular operation in the restoration of elbow flexion. 2. The technical difficulty of a particular surgical method and the postoperative results must be taken into consideration when selecting the right operation. 3. There is a need for controlled studies of the different methods. PMID- 3049274 TI - Bancroftian filariasis in the Igwun basin, Nigeria: an epidemiological, parasitological, and clinical study in relation to the transmission dynamics. AB - A 12-months study on bancroftian filariasis was carried out in the Igwun basin, Nigeria. A total of 1,418 individuals (768 males, and 650 females) were examined for microfilaremia and clinical filarial stigmata. There were 14.3% and 11.1% male and female point prevalence rates, respectively, and an overall prevalence of 12.8%. Prevalence rates and microfilarial density increased with age. The highest mff density of 35 mff/20 ml blood occurred in the 40-49 year old male individuals. Disease rates of 55.5 and 65.3% were recorded for males and females respectively. Chyluria (9.3% males, 16.7% females), hydrocele (17.8%), elephantiasis (15.9% males, 29.2% females), and enlarged groin glands (16.4% males, 19.4% females) were the major clinical signs, all associated with microfilaremia. Anopheles gambiae and Cules pipiens were the principal vectors. The estimated mean daily, weekly, and monthly per capita biting densities were 26, 161, and 753 respectively. The overall infection rate of mosquitoes was 22.3%, with a mean mff density of approximately 5 mff/mosquito. These vector parameters were indicative of active transmission in the area, and may be responsible for the high prevalence of infection, the diversity of clinical signs, and high morbidity rates. PMID- 3049275 TI - Rapid serological diagnosis of tick-borne encephalitis by means of enzyme immunoassay. AB - A system of IgM-capture EIA made up from Czechoslovak immunopreparations (SEVAC) was developed for a rapid serological diagnosis of tick-borne encephalitis (TBE). The method was tested on clinical material. The total IgM antibody titres were detected using pig antiserum and the selection of specific IgM antibodies was made with TBE antigen with following indirect way of detection. The antibody analysis made by means of this method is sensitive and fully conforms to the clinical picture of the disease. PMID- 3049276 TI - Identification of immunotoxic effects of chemicals and assessment of their relevance to man. AB - Immunotoxicity is defined as the adverse effects of foreign substances (xenobiotics) on the immune system. Two types of effects are possible: immunosuppression (which may result in an increased susceptibility to infection or to the development of tumours) and immunopotentiation (which may manifest as an allergy or as autoimmunity). There is, as yet, little evidence that well controlled occupational exposure to industrial chemicals has led to clinically significant immunosuppression. In contrast, a number of industrial chemicals have been shown to cause immunopotentiation in exposed populations, producing occupational asthma and contact dermatitis and possibly autoimmunity. In experimental models, immunosuppression (usually assessed by in vivo or in vitro immune function tests) has been induced by a wide range of chemicals but there are a few reports of the immunosuppression leading directly to an increased susceptibility to infection or to the development of tumours. Predictive experimental models are available for type IV allergic reactions, but the identification of chemicals that have a potential to cause other types of allergy or autoimmune reactions requires further research and the development and validation of new animal models. It is considered that routine subacute and chronic toxicity studies should include a full gross and histopathological assessment of the lymphoid organs to more accurately detect the potential of a chemical to cause immunotoxicity. Should such studies indicate that a substance has affected the immune system directly, an assessment of overall immune competence and function tests may be necessary using dose levels below those which cause frank toxicity. However, precise interpretation of immune function tests in terms of their relevance to human health requires an improved understanding of the extent of the functional reserve of the immune system. A strategy for assessing immunotoxicity in exposed human populations demonstrates a need for reliable clinical assessment, accurate medical record-keeping, an environmental and biological monitoring for levels of contaminating chemicals and the judicious use of well-validated immune function tests. PMID- 3049277 TI - Methylxanthines: toxicity to humans. 1. Theophylline. AB - While there are several comprehensive reviews on the toxic effects of theophylline, caffeine and theobromine in animals, data on the toxicity of these methylxanthines in humans have not been extensively reviewed in one document. This question will be addressed in a series of three papers. This paper provides an overview of the human toxicity of theophylline. Only pertinent and recent information on theophylline toxicity is summarized. In addition, some information regarding the use and benefits of theophylline, the mechanism of its effects and factors that affect variability in its clearance and half-life is also provided. Some problems in the analytical methodology of theophylline, problems that may be responsible for the controversy in the reported dose-response effects, are critically reviewed. PMID- 3049278 TI - Speaking fundamental frequency characteristics of normal Swedish subjects obtained by glottal frequency analysis. PMID- 3049279 TI - [Expansion and relapse]. PMID- 3049280 TI - [The femoropatellar pain syndrome. Clinical and radiologic diagnosis]. PMID- 3049281 TI - [Ultrasound imaging of the shoulder. Diagnostic value and therapeutic consequences]. PMID- 3049282 TI - [Applications and uses of static and dynamic measurement of posture (posturography)]. AB - Posturography is a very useful clinical instrument to quantify, analyze, and document postural sway and to differentiate patterns of instability that are typical for the different types of lesions encountered within the cerebellum and to differentiate cerebellar ataxia from spinal ataxia. An objective method of measurement like this is most valuable for follow-up studies and the surveillance of therapeutic efficacy. The method of recording short, medium, and long latency reflexes in leg muscles after a sudden tilt of a movable platform toe-up is not only totally atraumatic and fast, but more importantly it gives significant hints towards the possible site of a central lesion in the sensory-motor system. PMID- 3049283 TI - [Oropharyngeal dysphagia in neuromuscular diseases--differential diagnosis, examination procedure and therapy]. AB - After a synopsis of brain stem structures participating in swallowing, the main neurologic diseases are presented as a part of which swallowing disorders may occur: central and peripheral nervous system, motor end-plates and muscles can be involved. The sequence of clinical examination is described with special reference to findings in mouth, pharynx and larynx as well as to findings in high frequency cinematography. After description of normal swallowing pre-, intra- and postdeglutitive mechanisms of aspiration are shown. This distinction has proved useful for practical reasons. Finally we give a survey of therapeutic possibilities reported in recent years in literature and developed in our hospital, divided into indirect (facilitation of orofacial and laryngeal voluntary muscles) and direct therapeutic strategies (strategy of compensation during feeding). In connection with this therapeutic possibilities applied during pre-, intra- and postdeglutitive phase, depending on neuromuscular disturbances, are also described. PMID- 3049284 TI - The pathology of mitral valve prolapse. AB - The gross criteria for diagnosing prolapsing mitral valve are: 1. interchordal hooding of the involved leaflets, 2. hooding or doming of leaflets towards the left atrium, 3. elongation of the involved leaflets resulting in an increase in valve area, 4. dilatation of the valve annulus in patients with severe mitral regurgitation. The posterior leaflet is most frequently affected. The involved leaflets, in general, are thickened, soft, greyish white and have a smooth atrial surface. Chordae tendineae are described as elongated, tortuous and attenuated or thinned. Deviations from normal chordal insertion have recently been observed which possibly appear to represent the underlying abnormality. Microscopic findings include significant thickening of the spongiosa and the fibrosa, changes in dense collagen fibers in the atrialis layer, occasionally, with fibrin platelet deposits. Histochemical characterization of changes in the spongiosa may also be helpful in the diagnosis. Ultrastructurally, there may be changes in collagen and elastic fibers as well as myxoid areas. On comparison of findings in surgically-removed mitral valves with those of control specimens from autopsy patients with no cardiac abnormalities, the length of the anterior and posterior leaflet as well as the annular ring diameter was larger in the valves with prolapse. Two-dimensional echocardiography accurately assessed leaflet length when compared to morphologic measurements, however, the annular diameter during systole or diastole was smaller. In patients with mitral regurgitation requiring surgery, mitral valve prolapse is the most common cause. Annular ring dilatation and chordae tendineae rupture appear to contribute substantially to incurrence of the mitral regurgitation. The heart weight is increased in the majority of patients with symptomatic mitral valve prolapse but normal, however, in those without symptoms. The most frequent complication of mitral valve prolapse is mitral regurgitation with or without congestive heart failure. Patients with redundant leaflets may be at high risk of sudden death. Young women with abnormal resting ECG, prolonged Q-T interval, family history of sudden death or complex ventricular arrhythmias may also be at a greater risk of sudden death. The incidence of infective endocarditis appears higher in those with redundant than in those with nonredundant valves. The incidence of cerebral ischemic events is low.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3049285 TI - Mitral valve billowing and prolapse: perspective at 25 years. AB - The syndrome of primary mitral leaflet billow, with or without prolapse, is associated with myxomatous degeneration of the mitral valve apparatus, mainly the posterior leaflet, and the syndrome may be familial. It manifests clinically with an isolated nonejection systolic click (billow), a murmur of mitral regurgitation that is usually late systolic (prolapse), or a combination of murmur and click. Echocardiography identifies and assesses the extent of the billowing of mitral leaflet bodies but there are no specific echocardiographic criteria that can differentiate normal from pathological billowing. Similarly, a prolapsed leaflet is not detected echocardiographically when there is localized and mild failure of leaflet edge apposition but a more severely prolapsed, or flail, leaflet can be demonstrated and confirmed by that technique. Symptoms of the syndrome include anxiety, chest pain and palpitations. The resting electrocardiogram may show ST segment and T wave abnormalities. The majority of patients have a benign course and require reassurance only. Complications include systemic emboli, infective endocarditis, progression to severe mitral regurgitation, arrhythmias and, rarely, sudden death. Patients with prolapse of a leaflet edge are more likely to develop complications than those with only billowing of the leaflet bodies. Surgery, preferably valvuloplasty, is required for severe regurgitation and may also be indicated for potentially lethal tachyarrhythmias unresponsive to medical therapy. Mitral leaflet billow and prolapse may be secondary to, or associated with, many conditions. The prognosis is then principally that of the underlying disease of which ischemic heart disease and hypertrophic cardiomyopathy are the most important. PMID- 3049286 TI - Understanding mitral valve prolapse (MVP). AB - In asymptomatic or symptomatic patients with an audible click and late systolic murmur, mitral valve prolapse can be assumed to be present, the pathologic anatomical substrate of which is characterized by myxomatous changes in the mitral valve leaflets and collagen degeneration of the chordae tendineae. The conclusion that all persons with a systolic click have a diseased mitral valve and are at risk of complications is probably excessive. In the presence of an unequivocally-audible click and/or late systolic murmur, an echocardiogram for confirmation of the diagnosis is not necessary. If the auscultatory findings are uncertain, an M-mode recording and, because of its high sensitivity and specificity, a two-dimensional display from the parasternal long-axis view should be obtained. From the apical four-chamber view, false-positive findings may be incurred. A small percentage of patients with mitral valve prolapse have complaints which can be assumed attributable to disturbances in the neuroendocrine system (Tables 1 and 3). To what extent a relationship actually exists between autonomic dysfunction and mitral valve prolapse and whether or not this is coincidental, remains unclear. Treatment of the symptoms with anxiolytic drugs or beta-adrenergic receptor blocking agents is only indicated for disabling complaints if reassurance and psychological support are ineffective. Complaints of chest pain are atypical for angina pectoris. Supraventricular and ventricular arrhythmias may be observed (Table 3), the initial step in the management of which is to advise avoidance of irritants such as coffee, tobacco and emotional stress. Medical treatment is only indicated for hemodynamically-meaningful arrhythmias and in those patients in whom an increased risk of sudden death is present.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3049287 TI - [Surgical indications in asymptomatic internal carotid artery stenosis and in relation to heart surgery interventions]. AB - In the last 30 years, carotid endarterectomy has been employed on a wide-spread basis with the intention of providing surgical prophylaxis of stroke. Currently, however, there is no evidence available from prospective, randomized comparative studies indicating a clear superiority of surgical treatment versus medical treatment with respect to stroke prophylaxis or improvement in survival. Based on recent publications with sufficiently large patient populations, operative mortality appears to be about 1% and the rate of perioperative stroke about 3.4%. In those with symptomatic internal carotid stenosis, without surgery there is a 5% yearly risk of cerebral infarction such that carotid endarterectomy possibly appears warranted. In contrast, in association with asymptomatic internal carotid stenosis, that is, in the absence of any symptoms indicative of cerebral hypoperfusion, based on several recent prospective studies, the yearly rate of cerebral infarction is 1 to 2% and, consequently, less than that of the prophylactic surgical intervention. Additionally, carotid endarterectomy does not render complete protection against stroke and the follow-up curves for the respective treatments do not differ meaningfully, even during longterm observation. Accordingly, for asymptomatic internal carotid stenosis, the indication for surgery has not been clearly established. Among those with asymptomatic carotid stenosis, there may be a subgroup of individuals with high grade luminal obstruction or multiple vessel disease, who according to several studies, appear to be at a higher risk of subsequent complications even though this has not yet been confirmed by prospective, randomized studies.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3049288 TI - Influence of cycloheximide on acute diabetogenic effects of S-carboxymethylated human growth hormone in obese (ob/ob) mice. AB - Acute treatment of ob/ob mice with S-carboxymethylated hGH (RCM-hGH), a diabetogenic derivative of GH which lacks significant insulin-like and growth promoting activities, results in an increase in fasting plasma insulin and blood glucose levels and enhanced peripheral tissue insulin resistance. Plasma insulin level increases within 3 h after RCM-hGH is administered, whereas increased blood glucose concentration and enhanced peripheral tissue insulin resistance became evident 6 h after the hormone derivative is given. The lag period seen in the manifestation of these diabetogenic effects of RCM-hGH is consistent with the time required for gene expression. Therefore, the present study was undertaken to determine whether the above acute responses to the diabetogenic action of RCM-hGH would be expressed in ob/ob mice in which protein synthesis was blocked with cycloheximide. Female ob/ob mice were given either saline or cycloheximide (0.1 mg/g BW) ip and 1 h later were fasted and treated with either saline or 200 micrograms RCM-hGH ip. The mice were given a second injection of cycloheximide during the middle of the hormone treatment period to insure that protein synthesis remained blocked for the entire 6 h. In the animals not receiving cycloheximide, fasting plasma insulin level and blood glucose concentration were markedly elevated 6 h after the injection of RCM-hGH. Also, the GH derivative attenuated the ability of insulin added in vitro to stimulate glucose oxidation by adipose tissue segments isolated from the animals.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3049289 TI - Insulin-related materials in the nervous system of vertebrates and non vertebrates: possible extrapancreatic production. AB - Studies from multiple laboratories with a range of methods raised the possibility that insulin production occurs naturally at extrapancreatic sites. Part A covers the presence of insulin-related materials in organisms that do not have an endocrine pancreas, including unicellular prokaryotes and eukaryotes as well as multicellular non-vertebrate animals (insects et al.) and plants. Part B covers possible production of insulin by extrapancreatic tissues of vertebrates that are remote from a source of pancreatic insulin e.g. early chick embryos and mammalian cells in culture. Part C covers possible extrapancreatic insulin production in mammals in vivo. Each section ends with an outline summary with evidence in favor of and against the hypothesis. PMID- 3049290 TI - Isolation and partial characterization of insulin of the honeybee (Apis mellifica). AB - In the honeybee (Apis mellifica), insulin-like material was partially purified with acid ethanol extractions by a classic method for recovering insulin and following gel filtration on a Sephadex G-50 column. The preparations were characterized by their ability to cross-react with porcine insulin antibodies. Insulin-like biological activity was demonstrated using the insulin bioassay. Stimulation of glucose oxidation or lipogenesis was measured by isolated rat adipocytes. Insulin seems to be more widespread in invertebrates than was previously assumed. PMID- 3049291 TI - An insect brain peptide as a member of insulin family. AB - Amino acid sequencing of bombyxin (previously called 4K-PTTH) isolated from the heads of the silkmoth Bombyx mori has disclosed sequence homology of this insect neuropeptide with insulin. Immunohistochemistry using an antibody against a synthetic bombyxin fragment detected 4 pairs of immunoreactive neurosecretory cells in the dorso-medial region of the Bombyx brain. The same cells were reactive to bovine insulin antibody. PMID- 3049292 TI - An overview of insulin evolution. PMID- 3049293 TI - Bioactivity and pharmacokinetics of human proinsulin in comparison to human insulin after intravenous and subcutaneous injection. AB - The hypoglycemic actions of human insulin (1 IU/kg b.w.) and biosynthetic human proinsulin in about equimolar amounts were studied after intravenous and subcutaneous injection in rabbits. Blood samples were taken up to four hours after injection for the determination of blood glucose and immunoreactive levels of both insulin and human C-peptide. For the determination of human C-peptide, serum taken after proinsulin injection was divided into two fractions. One was examined directly by the human C-peptide radioimmunoassay and the other after incubation with a protein-A-sepharose coupled insulin antibody to find "free human C-peptide". Proinsulin in amounts equimolar to 1 IU insulin/kg b.w., exerted a 34% stronger hypoglycemic action after subcutaneous injection than after intravenous administration (area under curve estimation). Proinsulin induced hypoglycemia did not last longer after intravenous administration than that induced by intravenous insulin. Although subcutaneous proinsulin did not show the same maximum decrease of blood glucose compared to subcutaneous insulin, its action was significantly prolonged (up to 180 min). Specific measurement of free human C-peptide showed no evidence of conversion of proinsulin to insulin and C-peptide. PMID- 3049294 TI - Access to technology-assessment data needed. PMID- 3049296 TI - Medicare cost data too flawed to use? PMID- 3049295 TI - PROs use medical practice standards to assess quality. PMID- 3049297 TI - Satisfaction survey: not just a happiness index. PMID- 3049298 TI - AARP will fight further Medicare cuts. Interview by Jeffrey Finn. PMID- 3049300 TI - Nesidiodysplasia--a histologic entity? AB - Five cases of persistent neonatal hyperinsulinemic hypoglycemia (PNHH) were studied by a combined immunofluorescent and Feulgen technique under a computerized microscope which was used to measure the area and absorbance of beta cell nuclei. An increase of 16% was found in the nuclear area and absorbance of the beta cells in PNHH cases, in comparison with age-matched control cases. A smaller increase (9%) was found in nuclear area and absorbance of non-endocrine ductal cells in PNHH cases. In all PNHH and control cases, higher values of nuclear area and absorbance were found for beta cells in the islets of Langerhans than for nesidioblasts. The increase in average size of beta cell nuclei in PNHH can be used as a morphologic criterion for diagnosis in these cases. The increased absorbance of the nuclei stained with the Feulgen reaction is the morphologic expression of polyploidy, and has a correlation to the metabolic activity of beta cells in this disorder. We conclude that, in contrast to the normal postnatal developmental process of nesidioblastosis, nesidiodysplasia is a well-defined histologic entity, manifested by an increase of beta cell nuclear size and its DNA content. PMID- 3049299 TI - Recent progress in renal immunopathology. PMID- 3049301 TI - Treatment failure in Trichomonas vaginalis vaginitis. PMID- 3049302 TI - Detecting Chlamydia trachomatis by direct immunofluorescence using a Cytobrush sampling technique. AB - We compared two different methods of collecting endocervical samples for examination by direct immunofluorescence for Chlamydia trachomatis. A cervical Cytobrush gave better results than a dacron swab. Further studies should be performed to assess the value of alternative sampling methods to detect this organism. PMID- 3049304 TI - National planning for AIDS education. PMID- 3049303 TI - Comparative evaluation of particle agglutination test for antibody to human immunodeficiency virus. AB - A comparative evaluation of a new serological test for human immunodeficiency virus (HIV) was carried out. The test used the agglutination of gelatin particles coated with HIV antigen. It was found to be more sensitive than current enzyme linked immunosorbent assays (ELISAs) and more specific than the western blot test. Because of its simplicity, it promises to be of value, especially in developing countries. PMID- 3049305 TI - Psychophysical capacity modeling in frequent manual materials handling activities. PMID- 3049306 TI - Characterization of the human tumor and normal tissue reactivity of the KS1/4 monoclonal antibody. AB - The tissue and tumor distribution of the antigen recognized by monoclonal antibody KS1/4 was determined by a combination of immunoperoxidase techniques, flow cytometric analyses and solid phase enzyme-linked immunoassays. These data document that the KS1/4 antigen is expressed in many epithelial malignancies and normal epithelial surfaces suggesting that this antigen represents an epithelial/epithelial malignancy marker. PMID- 3049307 TI - A 3-D impedance method to calculate power deposition in biological bodies subjected to time varying magnetic fields. PMID- 3049308 TI - Faith and finance. PMID- 3049309 TI - Responses to high technology infertility treatment. PMID- 3049311 TI - Collaboration between nurses and physicians. PMID- 3049312 TI - Colonial heritage and nursing leadership in Trinidad and Tobago. PMID- 3049310 TI - Cognitive analysis of contraceptive behavior. PMID- 3049313 TI - Occupational health hazards for nurses--Part II. PMID- 3049314 TI - Molecular and functional properties of novel T cell subsets in C3H-gld/gld and nude mice. Implications for thymic and extrathymic maturation. PMID- 3049315 TI - T-cell lineages, repertoire selection and tolerance induction. PMID- 3049316 TI - Molecular indices of functional competence in developing T cells. PMID- 3049317 TI - Prethymic and intrathymic mouse T-cell progenitors. Growth requirements and analysis of the expression of genes encoding TCR/T3 components and other T-cell specific molecules. PMID- 3049318 TI - Human T-cell precursors: involvement of the IL-2 pathway in the generation of mature T cells. PMID- 3049320 TI - Naturally occurring mouse antibodies against T-cell-secreted chondroitin sulphate proteoglycan. AB - Stimulated T lymphocytes and certain T-cell hybridomas secrete molecules capable of inducing B-lymphocyte proliferation and differentiation. It has been shown recently that one such B-cell stimulatory factor is associated with chondroitin sulphate proteoglycan (CSPG) and was designated T-cell proteoglycan fraction, or T-PGF. We report here that mouse spleen cells cultured at high densities or stimulated with lipopolysaccharide (LPS) at low cell densities secrete antibodies directed against T-PGF. Such antibodies react primarily with the CSPG component of T-PGF and can inhibit the induction of plaque-forming cells (PFC) by T-PGF. By fusing high-density cultures of unstimulated mouse spleen cells with the myeloma P3 x 63AG8.653, several anti-T-PGF (CSPG) hybridomas were derived that exhibited activities identical to anti-T-PGF (CSPG) obtained from high-density spleen cell culture supernatants. The role that these spontaneously secreted autoantibodies may play in immunoregulation is discussed. PMID- 3049319 TI - Developmental status and reconstitution potential of subpopulations of murine thymocytes. AB - In this chapter we have summarized our view of the subsets of murine CD4- CD8- thymocytes which can be identified with a range of monoclonal antibodies. We have shown the division rate and turnover time of the main subsets and have listed what we know of the TcR gene rearrangement, and expression at the RNA and protein levels. We have been unable to completely segregate gamma delta-TcR-expressing cells from alpha beta-TcR-expressing cells by any of the markers we have used, although the proportions of the two receptor forms vary widely in the different subsets. Experiments involving intrathymic transfer of the CD4- CD8- subsets are described, which indicate that all the TcR- subsets of the CD4- CD8- thymocytes display some precursor activity and which suggest a progression of at least five stages through the TcR- subpopulations of CD4- CD8- cells. The earliest precursor is a Thy 1 low, HSA low, Pgp-1 high cell which has unrearranged C beta and is non dividing and which closely resembles the bone marrow prothymocyte. The later precursors are Thy 1 high, HSA high, Pgp-1 low, have rearranged C beta and are rapidly dividing. We tentatively conclude that none of the TcR+ CD4- CD8- cells are precursors of the major thymocyte subsets or of typical peripheral T cells, and we have found no evidence so far of separate precursors for the different mature subsets of thymocytes or peripheral T cells. PMID- 3049322 TI - Brain norepinephrine levels after BCG stimulation of the immune system. AB - Brain norepinephrine, dopamine and serotonin levels were determined in right and left hemisphere from female C3H/He mice 13 days after their immune system was stimulated by an intraperitoneal injection of bacillus Calmette-Guerin (BCG) (10(7) bacilli/mouse). Increased norepinephrine levels were observed in both hemispheres but significantly only in the right one. No concomitant variations in dopamine or serotonin levels were detected. Furthermore, norepinephrine levels in the right hemisphere appeared to be correlated with the ability of lymphocytes to proliferate after concanavalin A stimulation. The modulation of the immune system by the brain neocortex has been previously shown to be lateralized. Here we show that the information from the immune system towards the central nervous system also appears to be expressed in a lateralized manner. PMID- 3049321 TI - Immunohistochemical detection of deposits of eosinophil-derived neurotoxin and eosinophil peroxidase in the myocardium of patients with Chagas' disease. AB - An immunohistochemical study of eosinophil distribution in the inflammatory cell infiltrates of four different types of myocardial lesions associated with Chagas' disease--caused by Trypanosoma cruzi--showed larger numbers of these cells in areas presenting tissue necrosis and degeneration, most notably in patients with the most severe myocarditis from a histopathological stand-point. Using antisera specific for human eosinophil-derived neurotoxin or eosinophil peroxidase, we detected deposits of these secretion products on myofibres and in the interstitium of chagasic myocardium displaying necrosis and degeneration but rarely in other types of lesions. These deposits were not detectable in the myocardium of non-chagasic patients who had died from myocardial infarction (acute or in the scarring stage) or myocarditis secondary to bacterial endocarditis. When human eosinophil-derived neurotoxin was incubated with myoblast monolayers there was a significant cell injury, detachment and lysis. These effects were abrogated by yeast RNA, added as a competitive ribonuclease substrate, and inhibited by the ribonuclease inhibitor RNasin, suggesting that the ribonuclease activity of the eosinophil-derived neurotoxin was involved in the effect. These results suggest a link between eosinophil infiltration and necrosis in chagasic myocardial lesions and point to EDN, and perhaps other toxic eosinophil secretion products, as possible mediators of tissue damage. PMID- 3049324 TI - Monoclonal anti-gonadotropin releasing hormone (GnRH) produced by azotized GnRH preferentially recognise to native hormone. PMID- 3049323 TI - Immunobiology of human chorionic gonadotropin. PMID- 3049325 TI - In vitro effect of ions in presence of insulin and acetylcholine on glucose uptake or release by liver of domestic pigeons Columba livia. PMID- 3049326 TI - Gum guggul (Commiphora mukul)--the success story of an ancient insight leading to a modern discovery. PMID- 3049328 TI - Evaluation of integrated vector control measures on filariasis transmission in Pondicherry. PMID- 3049327 TI - Hepatoprotective activity of kutkin--the iridoid glycoside mixture of Picrorhiza kurrooa. PMID- 3049329 TI - Ultrasonic measurement of foetal parameters in normal pregnancy & in intrauterine growth retardation. PMID- 3049330 TI - Antibody dependent plaque enhancement by monoclonal antibodies against Japanese encephalitis virus. PMID- 3049331 TI - C.S.F. immunology in meningitis. PMID- 3049332 TI - Asymptomatic hematuria in childhood: causes and appropriate diagnostic studies. PMID- 3049334 TI - Modern management of nephrotic syndrome. PMID- 3049335 TI - Renal tubular acidosis: diagnostic work-up treatment and mechanisms of growth retardation. PMID- 3049333 TI - Pathogenesis of the hemolytic uremic syndromes: current concepts. PMID- 3049337 TI - Diagnosis ex juvantibus. Individual response patterns to drugs reveal hypertension mechanisms and simplify treatment. AB - Heterogeneity of response to antihypertensive therapy is a well-recognized clinical phenomenon. An agent that is antihypertensive in one patient may increase blood pressure in another or have no effect in a third. We believe that this variety of individual response to drug treatment can provide a new framework for the study of hypertensive subjects. Different patterns of response elicited by sequential trials of individual drugs with different mechanisms of action (diuretics, calcium channel blockers, alpha-blockers, beta-blockers, and converting enzyme inhibitors) should provide another means to classify hypertensive patients into biologically relevant groups. The documentation and analysis of this therapeutic heterogeneity in relation to renin profiling and to other physiological and demographic parameters may add a new dimension to the investigation of the pathophysiology of hypertension; it may serve as a basis for more appropriate stratification of participants in clinical trials and may ultimately contribute to a more rational approach to patient management. PMID- 3049339 TI - Fate of recombinant human renin administered exogenously to anesthetized monkeys. AB - Highly purified recombinant human renin (rh-renin), synthesized by Chinese hamster ovary cells, was labeled with iodine-125 and was given intravenously to pentobarbital-anesthetized common marmosets (Callithrix jacchus) to study the fate of the circulating renin. Specific anti-rh-renin antiserum was used to identify the 125I-rh-renin. Plasma disappearance of the exogenously administered 125I-rh-renin in marmosets (n = 6) showed two exponential components, with a half life of 12.1 +/- 1.9 minutes for the rapid component and 120.3 +/- 16.4 minutes for the slow component. The metabolic clearance rate was 1.17 +/- 0.26 ml/min/kg. Thirty minutes after the injection of 125I-rh-renin, 43.1 +/- 0.9 and 3.5 +/- 0.5% of the injected dose had distributed to the liver and the kidneys, respectively. With time, the accumulated 125I-rh-renin in the liver and kidneys decreased. The accumulation of 125I-rh-renin was less than 1% of the dose injected in other organs such as lungs, heart, spleen, adrenal glands, testes, and ovaries. Analysis of liver and kidney extracts by high performance liquid chromatography at 30 and 120 minutes indicated that immunoreactive 125I-rh-renin decreased with time and was accompanied by an increase in nonimmunoreactive degradation products of a low molecular weight. The incubation of 125I-rh-renin with monkey or human plasma at 37 degrees C did not degrade the labeled renin. Therefore, rh-renin was rapidly cleared from the circulation by the liver and the kidney. PMID- 3049340 TI - The discovery of nitric oxide as the endogenous nitrovasodilator. AB - Endothelium-derived relaxing factor (EDRF) is a labile humoral agent released by vascular endothelium that mediates the relaxation induced by some vasodilators, including acetylcholine and bradykinin. EDRF also inhibits platelet aggregation, induces disaggregation of aggregated platelets, and inhibits platelet adhesion to vascular endothelium. These actions of EDRF are mediated through stimulation of the soluble guanylate cyclase and the consequent elevation of cyclic guanosine 3',5'-monophosphate. EDRF has been identified as nitric oxide (NO). The pharmacology of NO and EDRF is indistinguishable; furthermore, sufficient NO is released from endothelial cells to account for the biological activities of EDRF. Organic nitrates exert their vasodilator activity following conversion to NO in vascular smooth muscle cells. Thus, NO may be considered the endogenous nitrovasodilator. NO is synthesized by vascular endothelium from the terminal guanido nitrogen atom(s) of the amino acid L-arginine. This indicates the existence of an enzymic pathway in which L-arginine is the endogenous precursor for the synthesis of NO. The discovery of the release of NO by vascular endothelial cells, the biosynthetic pathway leading to its generation, and its interaction with other vasoactive substances opens up new avenues for research into the physiology and pathophysiology of the vessel wall. PMID- 3049341 TI - Regulation of renin gene expression in hypertensive rats. AB - A carboxy terminal renin complementary DNA (cDNA) clone from rat kidney was isolated, characterized, and used as a probe for renin messenger RNA (mRNA) quantification in normotensive and hypertensive rats. RNA blotting analysis detected renin mRNA in control kidney and brain. Deoxycorticosterone acetate (DOCA) and high salt (1%) treatment of experimental animals resulted in a greater than 95% decrease in the content of renin mRNA in the kidney, as compared with values in control rats receiving 0.4% NaCl in their diet. In contrast, high salt (1%) treatment alone caused only a twofold decrease in kidney renin mRNA content, as compared with values in controls. DOCA and low salt (0.04%) or low salt (0.04%) treatment alone caused a 1.5-fold increase in the kidney renin mRNA content, as compared with values in control rats. These results indicate that DOCA and salt have a synergistic effect in depressing renin mRNA levels in kidney. Clipping of the left renal artery caused a threefold increase in the steady state level of renin mRNA in the ischemic kidney and a 0.5-fold decrease in the hypertrophied kidney. The data are consistent with the hypothesis that blood pressure and other stimuli regulate the expression of the renin gene in vivo. PMID- 3049336 TI - Recent developments in dialysis and transplantation. PMID- 3049338 TI - Volume (weight) loss and blood pressure response following thiazide diuretics. AB - Correlations were made between weight change (as an index of volume loss) and blood pressure (BP) reduction before and after hydrochlorothiazide treatment. A total of 343 patients with mild to moderate hypertension (95-114 mm Hg) received hydrochlorothiazide alone. The diuretic was titrated from 50 to 100 to 200 mg daily as needed until the diastolic BP fell below 90 mm Hg (goal BP) or side effects supervened. Of the 305 patients who completed the 10-week titration period, 65% attained goal BP. The effective dose of hydrochlorothiazide in 52% of these responders was 50 mg/day, and this was associated with weight loss averaging 1.58 kg. An additional 29% achieved goal BP with a similar degree of weight loss, but they required double the dose, or 100 mg/day. The remaining 19% of responders required significantly greater weight reductions averaging 3.14 kg to achieve goal BP, which necessitated hydrochlorothiazide, 200 mg/day. More blacks than whites attained goal BP despite similar degrees of weight loss in the two races. Plasma renin activity was initially higher in whites than in blacks and rose significantly more in blacks and whites requiring the greatest volume losses and the highest dose of hydrochlorothiazide to attain goal BP. Nonresponders had less weight loss than responders. Thus, diuretic dose requirements vary in different patients and are related either to different volume losses in response to a given dose or to different degrees of BP reduction in response to the same volume loss. PMID- 3049343 TI - Acute abdomen. AB - Acute surgical abdomen is the object of urgent surgical attention. The objective of emergency operation is to interrupt a process that has a steadily worsening prognosis on a scale of hours unless effective surgical treatment is rendered. There are basically three processes to address: free or incipient sepsis and peritonitis, gastrointestinal soilage, and hemorrhage. PMID- 3049344 TI - Emergencies of the biliary tract. AB - The traditional approach to urgent therapy of biliary tract disease has undergone significant change. Technologic advances now permit a nonoperative approach to acute cholangitis, acute gallstone pancreatitis and hemobilia. Acute cholecystitis continues to be treated surgically in most cases. The clinical use of such nonoperative therapy has been guided mainly by retrospective data. The precise indications and optimal timing for endoscopic and radiologic therapy and their relationship to traditional surgical therapy remains to be defined by careful prospective evaluation. PMID- 3049342 TI - Proteases released in organ culture by acute dermal inflammatory lesions produced in vivo in rabbit skin by sulfur mustard: hydrolysis of synthetic peptide substrates for trypsin-like and chymotrypsin-like enzymes. AB - The purpose of these studies was to identify some of the extracellular proteolytic enzymes associated with the development and healing of acute inflammatory lesions. Lesions were produced in the skin of rabbits by the topical application of the military vesicant, sulfur mustard (SM). Full-thickness, 1-cm2 central biopsies of the lesions were organ-cultured for one to three days, and the culture fluids were assayed for proteases with a variety of substrates. When compared to culture fluids from normal skin, the culture fluids from both developing and healing SM lesions had three to six times the levels of proteases hydrolyzing two synthetic peptide substrates: (1) t-butyloxycarbonyl-Leu-Gly-Arg 4-trifluoromethylcoumarin-7-amide(Boc-Leu -Gly- Arg-AFC, herein abbreviated LGA AFC), and (2) N-benzoyl-phenylalanine-beta-naphthyl ester (BPN). LGA-AFC is a substrate for trypsin, plasmin, plasminogen activator, thrombin, kallikrein, and the C3 and C5 convertases; BPN is a chymotrypsin and cathepsin G substrate. The culture fluids did not consistently hydrolyze four other synthetic peptide substrates or the proteins [14C]-casein and [14C]elastin. In order to determine the likely sources of LGA-AFCase and BPNase activity, we counted the number of granulocytes (PMNs), macrophages (MNs) and activated fibroblasts in histologic sections of developing and healing SM lesions, and we measured the levels of these enzymes in serum, in culture fluids of PMN and MN peritoneal exudate cells, and in culture fluids of two fibroblast cell lines. In SM lesions, serum and fibroblasts seemed to be the major source of LGA-AFCase, and serum alone the major source of BPNase. Tissue PMNs and MNs seemed to be only minor sources. The crusts of healing lesions, which were full of dead PMNs, seemed to be a rich source of both enzymes. In the SM lesion culture fluids, whether LGA-AFC and BPN were hydrolyzed by endopeptidases or only by exopeptidases could be determined by evaluating complex formation with alpha-macroglobulin proteinase inhibitors (alpha M). Endopeptidases, but not exopeptidases, are entrapped and inhibited by alpha M, because an internal peptide band in alpha M must first be hydrolyzed before molecular rearrangement (required for proteinase inhibition) occurs. The catalytic site of endopeptidases that are entrapped and inhibited by alpha M is known to remain active on (and reachable by) small synthetic peptide substrates such as LGA-AFC and BPN.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3049345 TI - Major complications of acute and chronic liver disease. AB - Many advances have been made in the understanding, diagnosis, and management of severe complications of liver disease. The pathogenesis of hepatic encephalopathy remains a challenge. Several toxins including ammonia, mercaptans, short-chain fatty acids, benzodiazepine-like substances, GABA-like substances, and impaired glutamatergic neurotransmission are at the top of the list of candidates. Use of the benzodiazepine antagonists is an experimental but promising new therapy in patients with hepatic encephalopathy. In patients with cirrhosis, spontaneous bacterial peritonitis (SBP) remains a common and highly lethal complication. The diagnosis of SBP is based on the polymorphonuclear cell count in the ascites and confirmed by culture of ascitic fluid. Early diagnosis and aggressive treatment has reduced mortality of SBP from greater than 90 per cent to 30 to 50 per cent. The appearance of cerebral edema in severe acute hepatocellular failure is associated with high mortality and conventional neurologic signs may be unreliable indicators of brain swelling. Current management of cerebral edema in fulminant hepatocellular failure may include early placement of an extradural sensor for continuous monitoring of intracranial pressure, so that short-term measures can be instituted making later liver transplantation safer. Coagulopathy remains a serious problem in patients with liver disease. Exchange plasmapheresis is a promising short-term adjuvant therapy. However, liver transplantation should be considered the definitive treatment for fulminant hepatocellular failure. The gastroenterologist often encounters multiorgan failure in patients with severe liver disease. Liver transplantation is now an important therapeutic consideration in almost every patient with severe, irreversible liver disease. Efforts should be targeted to early diagnosis of irreversible disease and coordination of patient care with a liver transplant center. PMID- 3049346 TI - Variceal hemorrhage. AB - Figure 2 is the algorithm followed in our institution for management of acute variceal hemorrhage. A small percentage of patients who present with active variceal hemorrhage will stop bleeding after gastric lavage alone. However, most patients require an intravenous vasopressin infusion at a dose of 0.4 units per minute, preferably combined with intravenous administration of nitroglycerin. Although glypressin and somatostatin may be associated with fewer side effects than vasopressin, the superiority of these drugs remains to be determined. Whether pharmacologic therapy succeeds or fails, most patients then proceed to endoscopic sclerotherapy. Sclerotherapy may be used as a temporizing measure in preparation for elective surgery or as a long-term, definitive treatment for prevention of recurrent hemorrhage. Balloon tamponade is reserved for patients who are bleeding too rapidly for effective sclerotherapy and for sclerotherapy failures in preparation for emergency surgery. Because recurrent hemorrhage frequently occurs after balloon deflation, a more definitive treatment (surgery or endoscopic sclerotherapy) should be planned for all patients who undergo balloon tamponade. Because operative risk is unacceptably high for patients with hepatic functional decompensation secondary to variceal hemorrhage, we believe that a policy of routine emergency surgery is unwise. Rather, emergency surgical intervention is reserved for the relatively small number of patients (15 to 25 per cent) who continue to bleed after nonoperative options have failed. Shunt surgery should be considered early in the course of patients with bleeding secondary to gastric varices and portal hypertensive gastropathy, both of which respond poorly to nonoperative measures. PMID- 3049347 TI - Emergencies in acid-peptic disease. AB - Perforation and hemorrhage are the major complications of peptic ulcer disease today. The significance of these two conditions is underscored by the more than 5000 deaths that resulted from these complications in the United States in 1979. A consideration of these two complications forms the basis of the present discussion. PMID- 3049348 TI - Intestinal obstruction. AB - Acute intestinal obstruction is an emergency that frequently requires operative intervention, either immediately or within several days. Because of the difficulty in distinguishing strangulation obstruction from simple obstruction, a philosophy of operating on all patients with intestinal obstruction has been advocated by many physicians. On the other hand, the approach of attempting tube decompression on most patients rather than immediate operation has also been advocated. Both of these approaches are extreme and somewhat simplistic in dealing with a problem that often is complex. Fluid and electrolyte replacement, intestinal tube decompression, and appropriate surgical management are crucial to the successful management of these patients. This text presents a summary of the pathogenesis and various etiologies of intestinal obstruction and offers some diagnostic and treatment guidelines that will assist the clinician in the management of his patients. PMID- 3049350 TI - Diverticulitis. AB - Diverticulitis represents a spectrum of clinical entities ranging from minimal pericolitis in the adjacent mesentery to uncontrolled intra-abdominal sepsis and septic shock. The presentation most often described is left lower quadrant abdominal pain, fever, chills, and left lower quadrant tenderness associated with a mass. Unusual presentations occur when infection tracts to distant locations. Diverticulitis is a common cause of intra-abdominal sepsis associated with high morbidity and mortality. The pathogenesis of intra-abdominal sepsis is not well understood, but likely involves circulating host inflammatory mediators. The role of computed tomography in the early diagnosis of diverticulitis is increasing and supersedes barium enema in the assessment of the extracolonic extent of disease. Also, computed tomographic-directed percutaneous drainage of intra-abdominal abscesses is, in most cases, as effective as surgical drainage. Predictably, the micro-organisms involved are representatives from the commensal flora of the lower gastrointestinal tract. These bacteria are usually sensitive to a wide range of antimicrobial agents that are effective against facultative and obligate anaerobic gram-negative bacilli. Surgical intervention is reserved for those individuals who do not respond to therapy, or for generalized peritonitis, uncontrolled sepsis, free viscus perforation, and fistulas. PMID- 3049349 TI - Lower gastrointestinal bleeding. AB - Lower gastrointestinal bleeding remains an important medical emergency. Most lower gastrointestinal bleeding is now known to come from angiodysplasia or diverticular disease. Accurate angiographic and colonoscopic diagnosis may lead to a better focus for treatment and improved survival. PMID- 3049351 TI - Emergencies in inflammatory bowel disease. AB - A review of acute emergencies in inflammatory bowel disease is presented. Caveats include prompt surgical drainage of loculated abscesses and aggressive management of bleeding or perforation. Adequate nutritional and immunologic assessment of all patients prior to surgery is paramount and has resulted in the widespread use of total parenteral nutrition. Aggressive medical therapy, particularly steroids, may be useful in "cooling down" cases of acute inflammation prior to surgery. This is particularly useful in Crohn's disease, in which recurrences abound and surgery has a significant degree of complication and recurrence. When urgent operation is necessary for acute ulcerative colitis, abdominal colectomy with ileostomy and preservation of the rectum is generally indicated. This will allow subsequent mucosal proctectomy and ileoanal anastomosis. When urgent colectomy is indicated for Crohn's colitis without rectal involvement, ileorectal anastomosis can be considered either as a primary or secondary procedure. If toxic megacolon is present, an initial trial of medical therapy is warranted in order to allow a single-staged operation to be performed electively. PMID- 3049352 TI - Gastrointestinal emergencies in the acquired immunodeficiency syndrome. AB - Opportunistic infection and neoplasia involving the gastrointestinal tract are common in AIDS patients. Life-threatening complications of this involvement may occur, requiring urgent diagnosis and therapy. We review the clinical presentation and approaches to management of AIDS-related gastrointestinal emergencies. PMID- 3049353 TI - The abdomen as a source of occult sepsis. AB - When a patient presents with sepsis and no clear etiology, the abdomen can hide a focus of infection and must be considered in the course of the evaluation (Fig. 1). There are certain groups of patients who do not exhibit the usual signs and symptoms of intra-abdominal infection and therefore constitute the population at risk for occult abdominal sepsis. These patients, for one reason or another, have an unreliable history or physical exam. Once intra-abdominal infection is suspected, certain basic laboratory and radiographic evaluations should be undertaken. Treatment delays are not tolerated and the performance of diagnostic tests when a laparotomy appears inevitable is not indicated. CT of the abdomen should not be used as a screening exam and should be reserved for those cases potentially having an infected fluid collection. If a thorough evaluation of the abdomen reveals a possible source, a measured medical and surgical approach can be undertaken, depending on the etiology. If no source is found, the question of a diagnostic laparotomy arises in certain cases (Fig. 2). This procedure should be reserved for those patients having some type of underlying abdominal surgery or pathology. Without a previous history of abdominal surgery or pathology, and with no other clinical evidence of intra-abdominal infection, a nondirected laparotomy can be safely performed when organ failure is not present but usually will not reveal a treatable lesion. Multiple organ failure may indicate the presence of a hidden abdominal source of infection; however, the window for successful surgical intervention may have already passed. Multiple organ failure does not mandate laparotomy when there is no clinical or radiographic basis for suspecting an abdominal source of infection. This is especially true if an alternative source of sepsis has been defined. PMID- 3049354 TI - Pancreatitis as a medical emergency. AB - Pancreatitis in its acute form, whether from alcohol or gallstones, can be a severe, devastating illness. Patients presumed to have the disease require aggressive fluid therapy and imaging of the pancreas by ultrasound and computed tomography (CT). They should be managed in an intensive care setting until the course of the illness is determined. Surgical intervention is reserved for cases in which extensive pancreatic necrosis or infection is demonstrated by CT scan. PMID- 3049355 TI - Perspectives on gastrointestinal infections in AIDS. AB - Gastrointestinal illnesses are among the most common and debilitating complication of infections with HIV, affecting 50 per cent to almost 100 per cent of AIDS patients in developed and developing countries, respectively. A number of factors including relevant modes of transmission, the environment, and immunosuppression conspire to determine which enteric infectious agents HIV infected persons acquire. In developed countries, transmission of a diverse spectrum of bacteria, viruses, and protozoa is facilitated by unprotected receptive anal intercourse and anal-lingual contact among homosexual men with multiple partners. In developing countries, where most HIV infections occur among heterosexual persons, waterborne and foodborne transmission are the principal modes of transmission of enteric organisms. The severity and duration of symptoms associated with enteric pathogens are determined by the host's immunologic response to the organism. Candida albicans often causes local mucosal disease but less often causes systemic infections in HIV-infected persons, likely because polymorphonuclear cell function is intact. The ability of AIDS patients to control infections with G. lamblia and C. jejuni is related to their ability to mount an antibody response to these organisms during infection. The virulence of the organism may also affect the clinical response to infection. Cryptosporidium causes diarrheal symptoms in both immunocompetent and AIDS patients, but illness is more severe and prolonged in the latter. Giardia lamblia and C. jejuni infections are associated with a range of clinical manifestations in both AIDS patients and HIV-seronegative persons, whereas CMV and possibly adenovirus appear to cause significant disease only among immunocompromised subjects. The availability of effective therapy is among the most decisive factors in determining the duration of enteric infections in AIDS patients. For example, Giardia lamblia may cause acute abdominal pain and diarrhea in HIV-infected subjects but prolonged infections with the parasite are uncommon because effective therapy is available. In contrast, infections with CMV and Cryptosporidium may be severe and chronic as available therapy is generally ineffective or only transiently effective. Awareness of these clinical, epidemiologic, immunologic, and therapeutic aspects of gastrointestinal illness in HIV-infected subjects should help to direct the diagnostic evaluation of these patients and to direct areas of research. PMID- 3049357 TI - Abnormalities of the intestinal immune system in AIDS. AB - Significant abnormalities in the distribution of lymphocyte subsets have been noted in the intestinal mucosa of ARC and AIDS patients. Abnormalities are most striking among the CD4 helper-inducer T-cell subset and likely reflects the direct effects of HIV infection of those cells. A deficiency in CD4-positive T cells in the intestinal mucosa potentially could result in significant abnormalities in mucosal immune function, including defects both in cell-mediated immunity and in the secretory IgA system. Such alterations in mucosal immunity and mucosal defense may render AIDS and ARC subjects susceptible to enteric infections and systemic infections in which the intestinal tract is the portal of entry. PMID- 3049356 TI - Gastrointestinal and hepatobiliary neoplasms in AIDS. AB - There is an increased frequency of Kaposi's sarcoma and non-Hodgkin's lymphomas in patients with AIDS. These "opportunistic malignancies" establish the diagnosis of AIDS in an HIV-positive patient and are associated with a high likelihood of gastrointestinal and hepatobiliary involvement. Kaposi's sarcoma is a multicentric cutaneous spindle-cell tumor that is associated with luminal lesions in at least 40 per cent of patients. Gastrointestinal KS is usually asymptomatic but may rarely bleed or obstruct. Treatment of KS with either radiation or chemotherapy can reduce tumor bulk, without affecting survival. Non-Hodgkin's lymphomas in AIDS are B-cell neoplasms composed of either noncleaved or blast cells, similar to those seen in Burkitt's lymphoma. The tumors are usually highly aggressive and present in extranodal sites in the majority of cases. Primary or secondary gastrointestinal involvement is frequent, with hepatic and rectal tumors being particularly common. Unlike KS, gastrointestinal lymphomas are usually symptomatic. Obstruction, perforation, and bleeding can occur in patients with luminal involvement. Jaundice due to hepatic infiltration or biliary obstruction may be seen. Treatment with chemotherapy is usually indicated because of the rapid progression of the tumor, although no prolongation of survival has been demonstrated. There may also be an increased incidence of Hodgkin's disease and anorectal neoplasia in patients with AIDS; however, these malignancies do not establish the diagnosis of AIDS in an HIV-positive patient. PMID- 3049359 TI - Gastrointestinal imaging in AIDS--abdominal computed tomography and ultrasound. AB - Radiologic imaging with computed tomography (CT) and ultrasound (US), when combined with guided fine-needle aspiration biopsy (FNAB) can diagnose a broad spectrum of gastrointestinal, visceral, and nodal abnormalities in AIDS. PMID- 3049358 TI - Endoscopic procedures in the AIDS patient: risks, precautions, indications, and obligations. AB - Gastroenterologists are frequently asked to perform endoscopy on patients with AIDS. A survey of university-based endoscopy units in the United States indicates that at the majority of these centers, endoscopic personnel are concerned about the risk of contracting AIDS and that these concerns have altered endoscopic practices. The actual risks are difficult to determine, but appear to be very low. Nevertheless, a variety of precautions are warranted to decrease the risk of transmitting AIDS virus or other pathogens to endoscopic personnel and patients. These precautions include the wearing of protective attire (gloves, gowns, masks, eye protection), careful handling of biological specimens, and adequate decontamination of equipment and surfaces. Indications for endoscopy in patients with AIDS are generally the same as those for other patients, but special diagnostic considerations include esophageal candidiasis, enteric Kaposi's sarcoma, cytomegalovirus infection, and a variant of sclerosing cholangitis. Patients with AIDS are entitled to the same consideration and medical care as other patients. This includes the performance of indicated endoscopic procedures. PMID- 3049360 TI - Gastrointestinal imaging in AIDS--luminal gastrointestinal tract. AB - Barium radiography of the gastrointestinal tract in patients with AIDS can assist in the evaluation of numerous problems including dysphagia, odynophagia, diarrhea, abdominal pain, symptoms of intermittent obstruction, and/or gastrointestinal bleeding. It can localize disease, evaluate complications, guide endoscopic biopsy, and monitor the progress of therapy. PMID- 3049362 TI - Gastrointestinal surgery in the AIDS patient. AB - The acquired immunodeficiency syndrome is associated with opportunistic infections and unusual malignancies that may affect the gastrointestinal tract. The number of patients affected by this condition is large and will continue to increase. The manifestations of the disease may mimic acute surgical conditions. In addition, standard surgical problems may occur in the AIDS patient. Diagnosis may be confounded by pre-existing symptoms and atypical presentations. An awareness and understanding of the disease processes distinctive to AIDS patients is essential for surgeons so that they may provide appropriate care to their patients and, at the same time, protect themselves and their fellow health care team members. PMID- 3049361 TI - The oral clinical features of HIV infection. AB - Patients with HIV infection and AIDS frequently experience one or more oral lesions at some time during the course of their disease. Recognition and management of these oral manifestations is an important component of AIDS care. PMID- 3049363 TI - Clinical approach to weight loss in the patient with HIV infection. AB - The loss of body weight is a common finding in the setting of AIDS or ARC. Weight loss usually heralds the appearance of an AIDS-associated infectious disease or neoplasm. Multiple mechanisms for catabolism exist, including decreased caloric intake, excessive caloric loss, and increased metabolic caloric consumption. Among the most serious implications of weight loss is the fact that many of the immune defects observed in persons with AIDS are exacerbated by protein-calorie malnutrition. There are myriad causes of weight loss in patients with AIDS and ARC. A systematic and thorough diagnostic approach guided by the patient's presenting symptoms often yields an etiology and therapeutic direction. PMID- 3049365 TI - Odynophagia/dysphagia in AIDS. AB - Odynophagia and dysphagia are common symptoms of treatable disorders of the esophagus in patients with AIDS. Esophageal candidiasis is the most frequent cause of these symptoms. In patients with AIDS or AIDS-related complex, thrush in combination with odynophagia or dysphagia almost certainly indicates the presence of esophageal candidiasis. Other causes of swallowing disorders in AIDS include opportunistic infection of the esophagus with herpes simplex virus, cytomegalovirus, or, rarely, cryptosporidiosis. Recently, ulcerative esophagitis in AIDS associated with unidentified viral-like particles has been described. Infrequently, Kaposi's sarcoma or lymphoma may involve the posterior pharynx or esophagus, respectively. Because Candida esophagitis is so frequently the cause of odynophagia and/or dysphagia in AIDS, it is suggested that in most cases, a therapeutic trial with an antifungal agent, like ketoconazole, may be appropriate before radiologic or endoscopic examination. Further investigation can be reserved for patients who do not respond to this trial or who have clinical evidence suggesting another esophageal disorder. Herpes simplex and cytomegalovirus esophagitis can be treated with antiviral agents, such as acyclovir and ganciclovir, respectively. Maintenance therapy with antifungal agents to prevent recurrent esophageal candidiasis may be beneficial, but the efficacy and cost effectiveness of this approach remain to be determined. Because of the increasing numbers of patients with AIDS, frequency of esophageal disorders, such as candidiasis, in these patients and the morbidity of these disorders, an expansion of clinical research efforts to determine effective treatment and prophylaxis for these disorders is warranted. PMID- 3049366 TI - Hepatobiliary abnormalities of AIDS. AB - Liver disease, although usually asymptomatic, is a frequent accompaniment of AIDS. Hepatomegaly and macrosteatosis are prevalent but non-specific findings. Evidence of remote hepatitis B virus infection is extremely common; however, the HBsAg carrier state, chronic active hepatitis, or cirrhosis occur no more frequently in AIDS patients than in the general population. Opportunistic intrahepatic infections (such as MAI, fungi, and CMV) or neoplasms (such as lymphoma or KS) usually reflect a disseminated process; liver involvement generally does not directly cause morbidity or result in death. Although biochemical liver tests are commonly elevated in the AIDS population, alkaline phosphatase has proved to be the most specific enzyme for infiltrative processes. Percutaneous liver biopsy has a high diagnostic yield, although the treatment options are currently limited. Acalculous cholecystitis and biliary tract obstruction have been recently described and probably result from CMV and/or cryptosporidial infection. Radiologic features of papillary stenosis and/or sclerosing cholangitis have been demonstrated. In contrast to hepatic parenchymal disease, these entities may be amenable to surgical or endoscopic therapeutic maneuvers. PMID- 3049364 TI - Infectious diarrhea in human immunodeficiency virus infection. AB - Gastrointestinal symptoms, particularly diarrhea, are common in patients with AIDS. Recent evidence indicates that enteric pathogens can be identified in most of these cases. Appreciating the clinical features caused by protozoal, fungal, bacterial, and viral pathogens will assist the clinician in effectively evaluating the gastrointestinal symptoms. Antimicrobial agents now are available for many of these pathogens. PMID- 3049368 TI - Evaluation and treatment of gastrointestinal tract hemorrhage in patients with AIDS. AB - Hemodynamically significant gastrointestinal tract hemorrhage is infrequently seen among patients with AIDS. During a 35-month period, we evaluated 37 AIDS patients with substantial gastrointestinal tract bleeding: 13 patients had upper gastrointestinal disease; 24 patients had colorectal disease. AIDS-associated lesions were identified as the etiology of the hemorrhage in 8 of 13 patients with upper and 9 of 24 patients with lower gastrointestinal tract bleeding. PMID- 3049369 TI - Diagnosis of vascular injury in children with supracondylar fractures of the humerus. AB - Four children with suspected vascular injury after supracondylar fractures of the humerus are presented. A noninvasive technique has been used in the diagnosis of vascular injury. A simple non-invasive method using the Doppler equipment connected to a spectrum analyser may exclude the need for arteriography and surgical exploration. PMID- 3049367 TI - Abdominal pain and anorectal disease in AIDS. AB - The diagnosis and treatment of abdominal pain and anorectal disease in AIDS patients are discussed. Emphasis is placed on the practical aspects of patient care. PMID- 3049372 TI - Bilateral, simultaneous, spontaneous rupture of the quadriceps tendon. A report of 3 cases and a review of the literature. AB - Bilateral, simultaneous, spontaneous rupture of the quadriceps tendon is a very rare injury. It usually occurs in the obese older patient (over 50 years). Often, patients are mistakenly treated for rheumatoid arthritis, mild strokes and even neurological paralysis. This diagnostic confusion can lead to a delay in treatment. There is often no history of significant trauma. The pathognomonic signs are a palpable suprapatellar gap and an inability to lift the straight leg. Surgical 'end to end' repair, even of delayed cases, yields satisfactory results. In none of our cases was protective additional tissue reinforcement or a pullout suture used. PMID- 3049371 TI - Crush syndrome complicating pneumatic antishock garment (PASG) use. AB - We present a case of severe compartment syndrome complicated by rhabdomyolysis and acute renal failure (crush syndrome) following the use of a pneumatic antishock garment (PASG) with survival of the patient. Review of the literature reveals one other similar case but without survival. The aetiology of the complication is discussed and recommendations for the safe use of the PASG are made. PMID- 3049370 TI - Closed pancreatic transection treated by Roux-en-Y anastomosis. AB - Two patients with closed pancreatic transection, were treated by Roux-en-Y pancreaticojejunostomy. The management of closed isolated pancreatic transection is discussed. PMID- 3049374 TI - Strain variation in phagocytosis of Cryptococcus neoformans: dissociation of susceptibility to phagocytosis from activation and binding of opsonic fragments of C3. AB - Phagocytosis of Cryptococcus neoformans is markedly influenced by the presence of a polysaccharide capsule. We examined activation and binding of C3 fragments to eight isolates of C. neoformans. All isolates were shown to have capsules by light and electron microscopy. These strains differed in susceptibility to phagocytosis by neutrophils. Yeast cells were opsonized by incubation in normal human serum. Five strains were resistant to ingestion, two strains showed intermediate levels of resistance to ingestion, and one strain was quite sensitive to phagocytosis. Yeast cells opsonized with heat-inactivated serum (56 degrees C for 30 min) neither attached nor were ingested by neutrophils. A quantitative estimate of the amount of C3 bound to the yeast cells was determined by use of normal human serum containing 125I-labeled C3. The results showed approximately 5 X 10(6) to 10 X 10(6) C3 molecules per yeast cell regardless of whether the yeast cells were sensitive or resistant to phagocytosis. Bound C3 was eluted from the yeast cells by treatment with 0.1 M NH2OH (pH 10), and the eluted fragments were examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions. Results of this analysis showed that little of the C3 was in the form of C3b, and there was substantial decay to iC3b, the inactive decay product of C3b. This pattern of decay was similar with all strains. Immunoelectron microscopy was used to assess the ultrastructural location of the C3 fragments bound to the yeast cells. C3 fragments were bound to the perimeter of the capsule regardless of whether the isolate was sensitive or resistant to phagocytosis. Thus, phagocytosis-sensitive and phagocytosis resistant isolates were similar with regard to the amount, molecular form, and ultrastructural location of C3 fragments bound to the cryptococcal capsule. These results further indicate that activation of the complement cascade is necessary but not sufficient for phagocytosis of the yeast cell. PMID- 3049373 TI - Immunization against tuberculosis: what kind of vaccine? PMID- 3049375 TI - Role of yeast cell growth temperature on Candida albicans virulence in mice. AB - Previous studies have suggested that yeast cell growth temperature may influence the relative virulence of the opportunistic dimorphic fungus Candida albicans. To test this possibility, mice were challenged with C. albicans yeast cells which were grown at either room temperature or 37 degrees C, and their survival was monitored daily. Mice which received room temperature-grown cells died faster. The interaction of glycogen-elicited polymorphonucleated neutrophils (PMNs) with C. albicans yeast cells grown at the two temperatures was examined, because PMNs have been shown to have a critical role in preventing development of candidiasis in normal individuals. In the absence of serum (i.e., nonopsonic conditions), more PMNs conjugated and engulfed C. albicans cells grown at room temperature than those grown at 37 degrees C. However, PMNs were less able to kill cells grown at room temperature than cells grown at 37 degrees C. Cells grown at room temperature also produced abundant germ tubes after engulfment and were thus more likely to escape killing by phagocytes. These results suggest that cells grown at room temperature are more virulent because they are less likely to be killed by phagocytes and are more likely to disseminate. The possibility that expression of cell surface hydrophobicity is involved in these events is discussed. PMID- 3049378 TI - Analytical variability of biological parameters of exposure and early effects. PMID- 3049379 TI - The impact of aspects of time and duration of exposure on toxicokinetics and toxicodynamics of workplace chemicals. PMID- 3049376 TI - Iron regulation of Serratia marcescens hemolysin gene expression. AB - The hemolytic activity of Serratia marcescens was examined as a function of iron availability. Restriction of iron by the nonmetabolizable chelator 2,2'-dipyridyl or the iron-binding protein transferrin produced a marked increase in hemolytic activity. The hemolytic activity of S. marcescens is determined by two adjacent genes, 5'-shlB-shlA-3', where shlA encodes the hemolysin which requires the ShlB protein for activity. A gene fusion between the promoter-proximal portion of shlA and phoA, the Escherichia coli alkaline phosphatase gene, was subcloned into a medium-copy-number vector, and the recombinant plasmid was introduced into S. marcescens. The expression of shlA was measured as a function of alkaline phosphatase activity, which increased threefold under iron-restricted conditions. Removal of the 5' noncoding region upstream of shlB in the fusion vector resulted in a 10-fold decrease in alkaline phosphatase activity under iron-sufficient conditions, with no effect of iron limitation on this residual activity. This suggested that the site mediating iron regulation of shlA expression occurs upstream of shlB. Consistent with this, we observed iron-regulated synthesis of the ShlB protein in Western immunoblots of isolated outer membranes. The hemolysin determinant was subsequently expressed on a medium-copy-number vector in fur+/fur isogenic strains of E. coli K-12, where a 10-fold-higher activity was observed in the mutant strain compared with the wild type. A sequence exhibiting some homology to the Fur-binding consensus sequence was identified upstream of the shlB coding region, overlapping the -35 region of a putative promoter. PMID- 3049377 TI - Variation in expression of antibody-dependent cell-mediated cytotoxicity in rodents with malaria. AB - Antibody-dependent cell-mediated cytotoxicity (ADCC) against mouse erythrocytes sensitized with immunoglobulin G was studied in mice with malaria. Spleen cells from mice had enhanced cytotoxic activity early in Plasmodium berghei infection but not later in the disease. Sera from infected animals and partially purified malarial immune complexes inhibited ADCC. In addition, ADCC was diminished in spleen cells from mice infected with the lethal variant of P. yoelii 17x compared with that in mice infected with the nonlethal variant. P. berghei-infected erythrocytes did not release 51Cr when incubated with effector cells unless the erythrocytes were sensitized with antibodies against normal mouse erythrocytes. PMID- 3049381 TI - Feasibility of composite workplace standards for chemical and/or physical workplace factors. PMID- 3049380 TI - The impact of physical and mental activity on toxicokinetics and toxicodynamics of workplace chemicals. PMID- 3049383 TI - Assessment of variability in individual exposure to metals. PMID- 3049382 TI - Towards a dynamic model of exposure, susceptibility and effect. PMID- 3049385 TI - Variability in toxicokinetics in exposure to solvents. PMID- 3049386 TI - Variability in toxicokinetics in exposure to metals. PMID- 3049384 TI - The impact of life style factors and consumption of pharmaceutical drugs on toxicokinetics. PMID- 3049387 TI - Screening models in occupational health practice for assessment of individual exposure and health risk by means of biological monitoring in exposure to solvents. PMID- 3049389 TI - The impact of nonwork-related impairments of organ functions on the toxicokinetics and toxicodynamics of industrial chemicals. PMID- 3049390 TI - The endocrine responses to general anesthesia. PMID- 3049388 TI - Enzyme induction: its relevance for internal exposure and health risks. PMID- 3049392 TI - Catecholamines in anesthetic and surgical stress. AB - The sympathetic nervous system is activated by a large variety of stresses and is important in the maintenance of cardiovascular and metabolic homeostasis. However, individual responses to sympathetic activation and circulating catecholamines vary widely. PMID- 3049391 TI - The endocrine responses to regional anesthesia. PMID- 3049393 TI - Anesthetic and surgical stress in the diabetic patient: carbohydrate homeostasis. PMID- 3049394 TI - The effects of acupuncture on operative pain and the hormonal responses to stress. AB - Variations in study results presented in this chapter may be attributed to a number of factors, including the population studied and type of hormonal assay used. Some researchers selected experimental groups from a population different from that used for the control group. It is unclear from the animal studies using nonacupunctured controls whether the controls received physical contacts with the experimenters comparable to those experienced by the experimental animals. The choice of acupuncture points used may also contribute to the inconsistency between experiments, since according to traditional acupuncture theory, points are not equivalent in their mediation of systemic effects. Duration of the acupuncture treatment, frequency of sampling, and time from treatment to last sample were not standardized. There is little consistency between studies even after adjustment for sampling time. Most of the statistically significant changes have been seen with electroacupuncture, and they appear to occur soon after onset of treatment, then disappear within 30 minutes of needle removal. However, the subjective changes in pain threshold have lasted several hours following needle removal. As an analgesic technique or adjuvant, acupuncture has been used most successfully in dental procedures and relatively superficial operations such as thyroidectomy; however, it has also been employed in more complex operations. The blood pressure and heart rate stability that have been observed with acupuncture analgesia have not been sufficiently explained and warrant further investigation, as does the possibility that acupuncture may be helpful in shock states characterized by increased stress hormone concentrations. PMID- 3049395 TI - Neonatal and pediatric stress responses to anesthesia and operation. PMID- 3049396 TI - Demonstration of IgE-sensitized mast cells in human heart and kidney. AB - We report the direct demonstration of IgE-bearing mast cells in human heart and kidney, as shown by immunohistochemical methods. This finding lends further support to the role of mast cells and IgE in the direct involvement of these two organs in immediate-type allergic reactions. PMID- 3049397 TI - Parenteral immunization with Shigella ribosomal vaccine elicits local IgA response and primes for mucosal memory. AB - The parenteral Shigella ribosomal vaccine (SRV), which previously was shown to protect guinea pigs and monkeys, has been compared with lypopolysaccharide (LPS) for its ability to induce a systemic and a local immune response. Injection of SRV caused a significant rise of the serum O antibodies of different classes and the appearance of IgA O antibodies in tears of guinea pigs and saliva and bile of monkeys. In guinea pigs, the local IgA response to parenteral SRV was much more intensive than that to feeding of high doses of LPS, while in monkeys it was nearly as high as that to challenge with a high dose of live pathogen. These data provide an immunological basis for the protective effect of SRV and are in disagreement with the widely accepted view of the inefficiency of parenteral antigens in stimulating mucosal immunity. The results are interpreted from the viewpoint of the role of ribosomes as a delivery system for the Shigella O antigen which provides high potency of SRV in stimulating local lymphoid tissue and makes it a good vaccine candidate. PMID- 3049398 TI - Theories of serial flow in intergenerational transfers. AB - Recent improvements in the economic position of the elderly population necessitate a more thorough understanding of the serial flow of economic supports from preceding to succeeding generations. Five theories which might extend our understanding of the decision rules employed in intergenerational transfers are discussed. They are: social exchange theory, the kin selection theory of altruism, human capital theory, social constructivist theory, and rational transfers theory. The underlying assumptions of these theories are presented and compared, and models of intergenerational transactions are developed. Comparative studies are recommended in order to gain deeper insights into the relative strengths and weaknesses of these different views of relationships between the generations. PMID- 3049399 TI - Depression, dementia, and social supports. AB - Recent literature on the relationships among dementia, depression, and social support was reviewed, with particular emphasis on the diagnostic differentiation of dementia and depression, and the role of these three entities in elderly individuals with cognitive impairment. Dementia-like symptoms arising in depression and the coexistence of dementia and depression are discussed. Research is necessary to determine more objective criteria for depression and dementia, to provide cognitive and psychiatric testing for elderly individuals, to clarify the diagnostic or prognostic value of the term pseudodementia, and to further elucidate relationships between depression, dementia, and social support. PMID- 3049400 TI - Process variables of the life review: counseling implications. AB - The concept of the life review as a developmental process is examined with three main objectives. First, the life review is placed within a developmental framework as a dynamic rather than static occurrence in which the individual is an active agent. Secondly, in order to clarify the possible process variables of the life review, three interactive stages are identified and elaborated. Finally, possible counseling interventions are suggested for practitioners working with elderly persons. PMID- 3049401 TI - abl oncogene expression in non-Hodgkin lymphomas: correlation to histological differentiation and clinical status. AB - Eight reactive lymphatic tissues, 166 cases of non-Hodgkin lymphomas (NHL) and 11 cases of multiple myeloma were investigated for expression of the c-abl protein using the poly-clonal anti-abl antibody 4411 and an indirect peroxidase technique. In selected cases the results were compared to those obtained with a second polyclonal and 2 monoclonal anti-abl antibodies. In 7 cases, Northern blot analysis of abl-mRNA was performed in parallel. In reactive lymphatic tissues, cells positive for the 4411 antibody were confined to the B-cell areas, i.e., to the mantle zone and parts of the germinal center. In NHL, a positive staining of the cell membrane was predominantly detectable in lymphomas putatively originating in the germinal center or mantle zone (in particular in centrocytic NHL), independent of their proliferative activity. Clinically, 7 out of 8 abl positive cases of chronic lymphocytic leukemia (CLL) had a more aggressive course of disease, whereas "progressive disease" occurred in only 7 out of 19 c-abl antigen-negative cases. When the clinical status of 78 patients with NHL and 11 patients with multiple myeloma was related to c-abl expression, c-abl-positivity was mostly confined to patients in advanced tumor stages [p less than 0.001 (NHL)]. PMID- 3049402 TI - Amiodarone in pregnancy. Case report and review of the literature. AB - A case of the use of amiodarone in pregnancy is reported and the literature on this subject reviewed. The data available to date show: there is no risk of teratogenic effects, the QT interval is prolonged during infancy (no associated arrhythmias noted), infant bradycardia occurs and should be monitored and thyroid function can be affected and should be monitored at birth. If fetal electrocardiographic monitoring is performed before and during labour and after birth, and thyroid function is assessed, then, to date, there does not appear to be any significant contraindication to the use of amiodarone during pregnancy. In view of the potential side effects, however, the use of amiodarone should be restricted to arrhythmias which are life-threatening or not controlled by conventional therapy. PMID- 3049403 TI - Does acute-phase beta-blockade reduce mortality in acute myocardial infarction by limiting infarct size? AB - The mechanism by which early intervention with beta-blockers reduces mortality in acute myocardial infarction is unclear. Therefore the effects of intravenous, followed by oral, metoprolol on indices of infarct size were studied in a double blind fashion with a median delay of 6.75 hours from onset of symptoms. In 129 patients peak enzyme release and QRS score on the electrocardiogram were assessed, while myocardial perfusion score on thallium-201 scintigraphy was studied in 45 patients. There was a close correlation between all the indices of infarct size. While the correlation coefficients did not appear to be influenced by metoprolol treatment, the slope of the regression was affected. Peak aspartate aminotransferase and lactic dehydrogenase were lower by 11 and 7%, respectively, in the metoprolol-treated group, but no reduction was noted in QRS score or in thallium-201 perfusion defect size in the actively treated group. Thus, it seems likely that early intervention with metoprolol in acute myocardial infarction reduces mortality, not by limiting infarct size, but by some other mechanism. PMID- 3049404 TI - Alcohol consumption as a public health problem in Papua New Guinea. AB - A 2-year alcohol study was undertaken for the Papua New Guinea Institute of Applied Social and Economic Research (IASER). This paper reports project findings bearing on general public health related to alcohol consumption: (1) Much trauma is alcohol related-particularly trauma caused by motor vehicle accidents. (2) Continued ingestion of nonbeverage alcohols--principally methanol--exacts a significant public health toll. (3) Patterns of drinking and national trends in beverage alcohol consumption suggest that alcoholic cirrhosis and cancer of the upper respiratory and upper digestive tracts will contribute to increased mortality among Papua New Guineans. PMID- 3049405 TI - Immunotherapy of patients with terminal-stage malignant tumors with an immunopotentiator OK-432--a comparison of SU-PS test-responding and nonresponding patients. AB - Terminal malignant tumor cases were treated with immunopotentiator OK-432. The absolute numbers of neutrophils and lymphocytes and different lymphocyte subsets were determined, and SU-PS and PPD skin tests were also performed to monitor the immunologic status of each patient. The SU-PS test changed to positive in one third of the patients 2-6 weeks after the start of therapy. The SU-PS responding patients showed an increase in lymphocytes and Leu II+ cells, and an elevation of the OK T4/T8 ratio 2 weeks later. In all patients from the responding group, the (OKIal+ - Bl+) cells decreased in the peripheral blood immediately after the positive change in the SU-PS test. In the SU-PS nonresponding group, the OKT4+ cells tended to decline with time, while the OKT8+ cells increased. That is, the OKT4/T8 ratio remained low throughout the test period. In the SU-PS responding group, OK-432 therapy prolonged the survival time. PMID- 3049406 TI - Rigor in health-related research: toward an expanded conceptualization. AB - When the research process is viewed not merely as a set of methods for data collection and analysis but, instead, as an integrated series of arbitrary choices made by the researcher, it quickly becomes apparent that the prevailing conceptualization of rigor is much too narrow; indeed, the way rigor is currently conceptualized may well be responsible for the many errors that are commonly made in the research process. An expanded conceptualization of rigor is presented, and its implications for research into some critical health issues are discussed in some detail. PMID- 3049407 TI - Causes of health and safety hazards in Canadian agriculture. AB - Agriculture remains one of the most hazardous occupations in the world, even in industrialized countries. One of the major differences between Canadian agriculture and most other sectors of the economy is that the vast majority of farmers are self-employed. Consequently their particular relations of production are expected to have an impact on the issue of work health and safety. After a review of the nature and extent of work accidents, deaths, illness, and injuries in farmers and farm workers, the article focuses on the causes of such hazards. These causes are analyzed with reference to individual, institutional, and structural factors. The author argues that institutional and structural factors seem to be of paramount importance in explaining the severity of farm health hazards. PMID- 3049408 TI - The development of Canadian nursing: professionalization and proletarianization. AB - In this article, the development of nursing in Canada is described in terms of three major time periods: the emergence of lay nursing, including organization and registration, 1870-1930; the move to the hospital, 1930-1950; and unionization and the routinization of health care, 1950 to the present. This development is viewed in the light of the orienting concepts of professionalization, proletarianization, and medical dominance (and gender analysis). This historical trajectory of nursing shows an increasing occupational autonomy but continuing struggles over control of the labor process. Nursing is now using theory, organizational changes in health care, and credentialism to help make nursing "separate from but equal to" medicine and to gain control over the day-to-day work of the nurse. Nursing can thus be viewed as undergoing processes of both professionalization and proletarianization. As nursing seeks to control the labor process, its occupational conflicts are joined to the class struggle of white-collar workers in general. Analysis of nursing indicates the problems involved in sorting out the meaning of concepts that are relevant to occupational or class analysis but which focus on the same empirical phenomenon. PMID- 3049409 TI - Racism, the National Health Service, and the health of black people. AB - Racism has been and is central to an understanding of the health of black people in Britain. Black people have played and are playing a central role in the National Health Service (NHS). Their role is, however, shaped by racism. Their experiences as consumers of the NHS are also shaped by racism--in terms of their treatment for both physical and mental health problems. In addition, their specific health problems such as sickle cell anemia have not received the attention they deserve. The NHS has become part of the internal control system of the British racist immigration system. The cuts in the NHS, and in other areas of the welfare state, since 1979 have created the conditions for increasing racial conflict on the one hand and for interracial class-based resistance on the other. PMID- 3049410 TI - Unethical behavior in an ethical industry? Critical coverage of the pharmaceutical industry, 1985-1986. PMID- 3049412 TI - Use of various animal erythrocytes for in vitro cultivation of Plasmodium berghei. PMID- 3049411 TI - A sensitive bioassay for serum cycloguanil using Plasmodium falciparum in vitro. PMID- 3049414 TI - Analysis of flow motion by the laser Doppler technique in patients with peripheral arterial occlusive disease. AB - Laser Doppler flux was measured at the forefoot in 12 healthy subjects and in 36 patients with different degrees of ischemia due to peripheral arterial occlusive disease. Two characteristic patterns of flow motion waves were observed: Large waves with a mean amplitude of 0.77 +/- 0.4 arbitrary units and a mean frequency of 3.03 +/- 1.0 c/min (0.051 +/- 0.02 Hz) and small waves with a mean amplitude of 0.21 +/- 0.1 arbitrary units and a mean frequency of 21.7 +/- 4.2 c/min (0.362 +/- 0.07 Hz). The prevalence of large waves tended to decrease with more advanced ischemia, whereas small waves occurred almost exclusively in ischemia and most frequently in severe cases. Large flow motion waves were enhanced during reactive hyperemia after arterial occlusion or appeared after peak flux had been reached. Time to peak flux or to vasomotion were reliable parameters for characterizing skin ischemia. The previously undescribed small flow motion waves might represent a compensatory mechanism involved in pathophysiology of ischemia. PMID- 3049413 TI - Repeated suicide attempts by the intravenous injection of elemental mercury. AB - The case of a patient who repeatedly injected himself intravenously with elementary mercury in suicide attempts is presented and the toxicological effects of this chemical form and route of exposure of mercury are examined. A review of the literature reveals that elemental mercury, when injected as opposed to inhaled, causes few of the effects typical of mercurialism; pleuritic chest pain was frequently reported, whereas renal and central nervous system involvement were less common. Evidence of premorbid psychiatric disturbances was found in ten of fourteen non-cardiac catheterization exposures to intravenous elemental mercury. Findings in our patient were consistent with these observations. One additional and noteworthy finding in our case was that documented deposits of elemental mercury in the right parietal lobe of the brain did not correlate with any specific deficits on neuropsychological testing. Consultation-liaison psychiatry plays an important role in the treatment and care of these complex patients. PMID- 3049415 TI - Impression cytology with transfer: an easy method for detection of vitamin A deficiency. AB - We described a new method to stain epithelial cells harvested by ocular impression. The cells are immediately transferred on a glass slide after the sampling; this transfer permits a very simple staining and easier reading by light microscopy. The results are compared with those obtained with the previous techniques and confirm ocular impression as a good help for vitamin A status determination. PMID- 3049416 TI - Metabolic and hormonal factors controlling food intake. AB - Food intake is controlled by stimuli acting at the pre- and postabsorptive levels. Preabsorptively, nutrients present in the stomach and intestine elicit signals contributing to meal-ending satiety. Neurally mediated signals and hormonal signals both seem instrumental in the production of this gastrointestinal satiety. Postabsorptively, metabolic receptors in the liver that sense the oxidation of metabolic fuels seem to play an important role in the control of post-meal satiety and meal initiation. These receptors are apparently connected with the brain by vagal afferents, because hepatic vagotomy eliminates the effects of various metabolites and metabolic inhibitors on food intake. Glucoreceptors in the hindbrain that sense the utilization of glucose are probably also involved in control of meal initiation. Finally, humoral factors reflecting the size of the fat depots may also function as feedback signals in the control of feeding. PMID- 3049417 TI - [Unconventional treatment methods in internal medicine]. PMID- 3049418 TI - [Medical and scientific experience as complementary guidelines of therapy]. PMID- 3049419 TI - [Phytotherapy in internal medicine]. PMID- 3049420 TI - [Plant preparations for immunostimulation]. PMID- 3049422 TI - [Homeopathy in internal diseases]. PMID- 3049421 TI - [Acupuncture in internal diseases]. PMID- 3049423 TI - [Dietary aspects of cancer diseases]. PMID- 3049425 TI - [Chinese and Western medicine--confrontation of 2 medical cultures]. PMID- 3049426 TI - [Cryptococcoma of the lung--a contribution to diagnosis and therapy]. PMID- 3049427 TI - [Heparin-induced thrombocytopenia and thrombosis]. PMID- 3049424 TI - [Alternative methods in peripheral vascular diseases]. PMID- 3049428 TI - Prostacyclin is the major prostaglandin synthesized by bovine retinal capillary pericytes in culture. AB - Prostaglandin synthesis by bovine retinal pericytes was investigated using high pressure liquid chromatography to separate and identify 3H-labeled prostaglandins released from 3H-arachidonic acid labeled pericyte monolayers. A dominant peak activity corresponding to 6-keto-PGF1 alpha was observed. This peak was eliminated when monolayers were pretreated with cyclooxygenase inhibitors and was augmented when monolayers were stimulated by the calcium ionophore A23187. Suspensions of pericytes and the cell-free media of monolayers incubated with arachidonic acid inhibited adenosine diphosphate-, collagen-, and arachidonic acid-stimulated platelet aggregation in a bioassay for prostacyclin-like activity. This inhibitory activity was unstable at room temperature. Cultures of 7.5 to 10 x 10(5) pericytes (7th passage near-confluence) released nanogram quantities of 6-keto-PGF1 alpha as measured by radioimmunoassay. These results are evidence that prostacyclin is the main prostaglandin synthesized by bovine retinal capillary pericytes in culture. Pericytes may influence the microcirculation via their production and release of this potent vasoactive arachidonic acid metabolite. PMID- 3049429 TI - Effects of laminin on attachment, growth and differentiation of cultured Y-79 retinoblastoma cells. AB - In vitro, as in vivo, the attachment, growth and differentiation of many cell types are dependent upon the availability of appropriate extracellular matrix (ECM) molecules. Here we have studied the effects of ECM components, including fibronectin and laminin on cultured Y-79 retinoblastoma cells. Both in 2 hr and in 3 day studies, the highest frequencies of attachment were seen with a laminin substrate (50 micrograms/35 mm culture dish). Attachment was significantly inhibited by specific anti-laminin antibodies. In longer studies of up to 1 week, laminin or fibronectin was added directly to the culture medium. Neither molecule significantly stimulated cell growth, but laminin continued to promote the highest frequencies of attachment (20% to 30% greater than any other substrate). Laminin exposure also caused morphological changes in Y-79 cells. Many cells became flattened and extended long, branching, neurite-like processes. These changes could be inhibited by inclusion of anti-laminin antibodies. Such studies may provide information about events occurring during normal eye development as well as about tumor cell attachment and growth. PMID- 3049430 TI - Substrate modulation of cultured rabbit conjunctival epithelial cell differentiation and morphology. AB - This study demonstrated that growth and differentiation of rabbit conjunctival epithelial cells could be promoted by such substrata as collagen gel, matrigel or various mixtures of collagen and matrigel in a defined culture system. On conventional plastic or glass culture, the conjunctival epithelial cells adopted a monolayer of small epithelioid cells in primary cultures. They became enlarged, squamoid and exhibited notable senescence upon subcultures. On collagen gel, cells formed an organized monolayer sheet with cuboid shape and cell polarity. On matrigel, cells formed globules with stratified appearance including the basal layer of the outer part of globule and the squamoid cells of the central part of globule. The epithelial origin of these cultures was verified by the positive immunostaining of anti-keratin monoclonal antibodies. Expression of mucin antigen was lost on plastic or glass culture, but promoted on collagen gel or matrigel, as demonstrated by staining with periodic acid Schiff's reagent and anti-mucin monoclonal antibody stainings. These results indicate that both collagen gel and matrigel can provide a permissive substrate environment for goblet cell differentiation. Furthermore, this unique phenotypic expression may be possessed only by a selective cell subpopulation. This culture system will allow us to further explore the mechanism by which the goblet cell differentiation is controlled. PMID- 3049431 TI - Evolution of high-dose cisplatin. AB - High-dose cisplatin therapy, defined as 200 mg/m2/course, is currently undergoing extensive clinical trials in a variety of solid tumors. The reduction of the incidence and severity of cisplatin-induced nephrotoxicity has led to clinical trials of higher doses of cisplatin. By maintaining nephrotoxicity to acceptable levels, dose response relationships have shown increased efficacy of cisplatin therapy. However, new dose-limiting toxicities, primarily severe neurotoxicity and myelosuppression, have prevented further dosing increases. The following review will trace the evolution and the current status of high-dose cisplatin therapy. In addition, a summary of the dose-limiting non-renal toxicities and their relationship to pharmacokinetics and dosing schedules will be discussed. PMID- 3049432 TI - In vivo and in vitro investigations on biological effects of aromatic bis-(2 chloroethyl)amino-bisphosphonic acids, new agents proposed for chemotherapy of bone tumors: cytostatic activity in rat osteosarcoma; toxicity and genotoxicity in liver and bone marrow; mutagenicity in S. typhimurium. AB - Two new aromatic bis-(2-chloroethyl)-amino derivatives (BCMP and BAD) which are linked to osteotropic bisphosphonates were investigated for their therapeutical efficacy in rat osteosarcoma. Furthermore their genotoxic potential in vitro was determined in S. typhimurium and in mammalian cells. Finally, parameters for toxicity and genotoxicity were determined in liver and bone marrow cells following in vivo treatment. It was shown that BAD was of higher therapeutic effectiveness than BCMP. Both compounds induced approximately a two fold increase of his+ revertants in S. typhimurium TA1535 following metabolic activation by subcellular liver fractions. Both compounds also induced amplification of SV40 DNA in SV40 transformed cells (CO631). This endpoint may be of importance for acquired resistancy of cells during therapy. DNA-single strand breaks were induced by BCMP but not by BAD in liver cells and CO631 cell line. Following in vivo treatment BCMP was of higher genotoxic activity in liver cells than BAD. In comparison, genotoxicity of both compounds was much lower in bone marrow cells than in liver cells. BCMP was again more potent than BAD in inducing DNA single strand breaks, whereas BAD was more toxic. The higher therapeutic efficacy of BAD together with its lower genotoxic properties makes this compound superior to BCMP as a candidate for applied chemotherapy in humans. PMID- 3049434 TI - Diagnostic imaging in clinical cancer management. Metastases from unknown primary tumors. PMID- 3049433 TI - Clinical trial of iproplatin (cis-dichloro-trans-dihydroxy-bis-isopropylamine platinum IV, CHIP) in patients with advanced breast cancer. AB - Twenty-five women with advanced breast cancer were treated in a phase II trial of iproplatin 275 mg/m2 administered intravenously every 4 weeks. All patients had measurable or evaluable indicator lesions, and had undergone treatment with no more than one previous chemotherapy regimen, including adjuvant chemotherapy. Two of the twenty-four evaluable patients (8%) experienced major therapeutic responses. One patient had a complete regression of pulmonary nodules lasting 18+ months; another had a partial regression of metastatic disease in the liver (4 months). The inevaluable patient was ineligible for the study because of previous radiation to the indicator lesions on her chest wall; nonetheless, she experienced a 10 month partial regression of those nodules. Myelosuppression was generally dose limiting; thrombocytopenia was more profound, but leukopenia was more prolonged. Nausea, vomiting, diarrhea, and general malaise were prominent toxicities, and led to discontinuation of therapy in 4 patients. Iproplatin has limited activity in previously treated women with advanced breast cancer. PMID- 3049435 TI - Urological complications in 454 renal transplants. PMID- 3049436 TI - Crohn's disease in Dublin in the latter half of the nineteenth century. PMID- 3049437 TI - The psychology of visual perception in Ptolemy's Optics. PMID- 3049438 TI - Genetics in China. The Qingdao Symposium of 1956. PMID- 3049439 TI - Gleanings from an Arabist's workshop. Current trends in the study of medieval Islamic science and medicine. PMID- 3049440 TI - The ontogeny of Sewall Wright and the phylogeny of evolution. "Sewall Wright and Evolutionary Biology". By William B. Provine. Essay review. PMID- 3049441 TI - A missing link in the evolutionary synthesis. "Factors of evolution: the theory of stabilizing selection". By I. I. Schmalhausen. Essay review. PMID- 3049442 TI - Proteolysis in human erythrocytes is triggered only by selected oxidative stressing agents. AB - Human erythrocytes have been treated with different agents producing oxidative stress. Diamide, tetrathionate, chromate, cystamine and iodate preferentially influenced the cellular redox state by oxidation of free and protein thiol groups leaving the redox state of hemoglobin virtually unaffected. None of these compounds was able to stimulate the proteolysis. By contrast, phenylhydrazine, nitrite, hydrogen peroxide, ter-butylhydroperoxide, cumene hydroperoxide and copper-ascorbate caused a noticeable oxidation of hemoglobin to methemoglobin. These latter agents, except nitrite and copper-ascorbate, triggered proteolysis. Identical results have been obtained in a ghost-free hemolysate. The fraction containing the proteolytic activity was isolated from hemolysate and tested on native or oxidant-treated hemoglobin. The proteolysis was stimulated by all agents able to produce methemoglobin. It is concluded that proteolysis correlated to an unbalance of cellular redox state. The results obtained with isolated and recombined fractions suggests that increased proteolysis does not depend on the removal of the effect of protease(s) inhibitor(s). Since all agents stimulating proteolysis are able to generate free radicals, it seems that protein breakdown is triggered by the direct effect of these intermediates on proteins (mostly hemoglobin) without the involvement of radical species produced in the membranes by action of organic hydroperoxides. In addition, since nitrite and copper ascorbate, which also oxidize hemoglobin by radical generation, are unable to stimulate proteolysis, it should be concluded that protein degradation induced by oxidative stress is a complex event induced only by specific agents. PMID- 3049443 TI - Transient global amnesia: definition and clinical phenomenology. AB - Based on personal observations and analysis of the most impressive literature, some considerations are made on transient global amnesia definition and clinical phenomenology. PMID- 3049444 TI - Post-traumatic transient amnesias. AB - Temporary memory disorders following closed head injuries are presented, with special remark on the cases of transient global amnesia (TGA) triggered by head trauma. The relationship with idiopathic forms of TGA is discussed. PMID- 3049445 TI - Transient psychogenic amnesia. AB - Features of transient psychogenic amnesia (TPA) are described and related states of psychogenic amnesia are outlined and discussed. A few remarks on possible therapeutic interventions are given. It is concluded that despite the existence of common attributes in various transient amnesic forms and in spite of the possibility of mixed symptomatologies, distinctive and characteristic features for most cases of TPA can be listed and relied on. PMID- 3049446 TI - Paul W. Gebauer: Hawaii's illustrious thoracic surgeon. PMID- 3049449 TI - On the origin of head pain. PMID- 3049448 TI - Update on cancer therapy. PMID- 3049447 TI - Hemoglobin H disease in Hawaii: update 30 years later. PMID- 3049450 TI - Hemiplegic migraine in pregnancy. PMID- 3049451 TI - Childhood muscle contraction headache: current issues in assessment and treatment. PMID- 3049452 TI - Public hospital strategic planning: does it differ from voluntary, not-for-profit hospital strategic planning? AB - One-hundred-eighty-six public and voluntary, not-for-profit hospitals were surveyed regarding their strategic planning processes, which were found to be more similar than different. Public hospitals, however, were less flexible, less formal, and more hesitant to adopt the latest strategic planning innovations. PMID- 3049453 TI - Survival of the hospital emergency department: strategic alternatives for the future. AB - Diverse and pervasive environmental forces are reshaping hospital emergency services as hospitals strive to respond to consumer preferences related to cost and convenience. Complacency can no longer serve as a standard operating procedure for hospital emergency departments competing against lower-priced, consumer-oriented, free-standing facilities. Strategic alternatives, a five-step strategy for survival and growth, and a projection of future models of hospital emergency services are examined. PMID- 3049454 TI - An additional role for the Health Physics Society. PMID- 3049455 TI - The Helen Q. Woodard symposium on radiation carcinogenesis and associated dosimetry. PMID- 3049456 TI - Evolving perspectives on the concept of dose in radiobiology and radiation protection. AB - Since the discovery of the x ray more than 90 y ago, the biological effects of radiation have been a subject of intensive and continuing study. At the outset, such study was severely hampered by the lack of a suitable method of dosimetry. More than a quarter of a century elapsed before the introduction of a quantitative system for measuring exposure, and another quarter of a century elapsed before the introduction of quantitative units of absorbed dose. In the meantime, the effects of a given dose had long since been found to depend on its distribution in space and time; that is, on the precise spatial and temporal patterns of energy deposition within absorbing tissues and cells. Study of the biological effects of radiation thus led to elaboration of the concept of dose, to take into account relevant microdosimetric parameters. Advances in ongoing research on the molecular mechanisms of radiation effects can be expected to result in further evolution of such coNcepts. PMID- 3049457 TI - Carcinogenesis--a synopsis of human experience with external exposure in medicine. AB - Studies in the 1980s of medically irradiated populations have increased our knowledge of radiation carcinogenesis. (1) Investigations of prenatal x-ray exposures, especially in twins, provide evidence that very low doses of ionizing radiation may cause cancer in humans. (2) Fractionated doses appear as effective as single exposures of the same total dose in causing breast cancer, but seem less effective for lung cancer. (3) Excess breast cancers can occur among women exposed under age 10, indicating that the immature breast is susceptible to the carcinogenic action of radiation. (4) Moderate doses on the order of 1 Gy to the brains of children can cause tumors later in life; moderately high doses to the skin can cause cancer when followed by frequent exposure to ultraviolet light. (5) Radiotherapy for cervical cancer can increase the rate of subsequent leukemia with the best fitting dose-response functions including a negative exponential term to account for cell-killing. (6) Low-dose exposures of about 10 cGy may increase the risk of thyroid cancer. (7) Second cancers following radiotherapy for a variety of cancers occur primarily among long-term survivors. (8) Radiotherapy may not significantly increase the risk of leukemia following childhood cancer, whereas chemotherapy with alkylating agents is a major risk factor. (9) Bone cancer occurs after high-dose radiotherapy for childhood cancer, but children with retinoblastoma are not more susceptible to radiation-induced disease than children with other malignancies. (10) High-dose external beam therapy can cause thyroid cancer, whereas high-dose radioactive 131I may not. (11) Studies of cervical cancer patients indicate that the risk of radiation induced second malignancies follows a time-response model consistent with a constant multiplication of the underlying background incidence, i.e. a relative risk model seems to hold for projecting risks forward in time. PMID- 3049458 TI - [Immunologically induced fertility problems in man]. PMID- 3049459 TI - [Collaboration and specific role of the urologist and gynecologist facing a hypofertile couple]. PMID- 3049461 TI - [Andrews bacterid (pustulosis palmaris et plantaris) in orthopedics. Description of 2 cases and literature review]. PMID- 3049460 TI - [Proton spin tomography in testicular tumors]. PMID- 3049462 TI - [Orthotopic hepatic transplantation]. PMID- 3049463 TI - The hepatic artery in orthotopic liver transplantation. PMID- 3049464 TI - Cases of abnormal human hemoglobin produced by de novo mutation. PMID- 3049465 TI - [Hematopoietic growth factors: towards a therapeutic revolution]. PMID- 3049466 TI - [Magnetic resonance in pediatric research and clinical practice. I. What can we expect from this new method?]. AB - Nuclear Magnetic Resonance (NMR) was first observed over 40 years ago and has recently also entered the field of human medicine. It currently attracts increasing attention from biologists and clinicians alike, and the scope of its different applications is in a phase of explosive development. Two principle developments of the MR method are taking place over the recent years and are of special interest for pediatricians and neonatologists. One involves the possibility of obtaining images from any part of the human body, somewhat similar to those obtained with computer tomography (CT), but without any radiation hazard. Today clinicians are most familiar with this mode of MR application. The other development tries to adapt the MR method of elucidating the structure of molecules used in physics, molecular biology and organic chemistry for applications in medicine, allowing to study metabolism in vivo under non-invasive conditions. Again, such studies pose no health hazards and are, therefore, applicable to neonates and small infants. They will enhance our understanding of metabolic processes during normal development and disease, especially in organs like the brain, where biopsies are virtually impossible. Recent developments combine the two methods mentioned above, in order to obtain morphological as well as metabolic information from the same organ at the same time, which may provide even better insight into pathophysiological mechanisms and their response to therapeutic measures. This article attempts to give an overview to the medical researcher, the clinician, and especially the pediatrician and neonatologist of what MR is and what we can expect from it. PMID- 3049467 TI - Congenital toxoplasmosis presenting as massive neonatal ascites. AB - A preterm infant with isolated transudative ascites caused by Toxoplasma gondii is described. In the absence of obvious congenital malformations, toxoplasmosis should be considered in the differential diagnosis of fetal and neonatal non immune ascites. PMID- 3049468 TI - Ultrasonographic manifestations of liver abscesses; an experimental study. PMID- 3049469 TI - Production of therapeutic polypeptides through Escherichia coli: a perspective. PMID- 3049471 TI - [Follow-up treatment and after care following operations to improve hearing]. AB - Postoperative treatment is very important for the success of an operation to improve hearing, not only in the immediate postoperative phase, but also in the long term. In most cases this medical treatment can be delegated by otolaryngologists to office practice, providing certain prerequisites are fulfilled. We describe indications and technics for postoperative treatment after stapedectomy and tympanoplasty. PMID- 3049470 TI - Immunohistochemical detection of antibody in tissue sections of non-perfused and ex vivo-perfused organs using a tetrazolium alkaline phosphatase substrate. AB - We have used nitroblue tetrazolium (NBT) as a color reagent to localize antibody bound alkaline phosphatase in frozen tissue sections. In the method described, NBT is reduced to a stable black diformazan reaction product that contrasts well with nuclear counterstains such as hematoxylin and stands out strongly in black and white photographs. We have found NBT to be a suitable color reagent for the alkaline phosphatase: anti-alkaline immunohistochemical technique. The reaction product also contrasts well with fast red and can therefore be used as second reagent for two color immunoenzyme studies. In this report, we describe a novel two color immunoenzyme method to assess the ex vivo binding of antibodies against Class II histocompatibility antigens in whole organs connected to a perfusion circuit. PMID- 3049472 TI - [Otologic aspects of diving]. AB - Ear diseases are the most common of all occupational diseases of diving. Otitis externa is the most frequent and troublesome infection in divers especially when the environment is humid. During compression, failure to equalize the pressure of the air-filled cavities surrounded by bone (middle ear, sinus) deprives the middle ear (or sinus) of aeration. Middle ear barotrauma is the most common barotrauma encountered in divers while external ear barotrauma (reversed ear) and inner ear barotrauma (with rupture of the round or oval window) are less common. Decompression sickness (Caisson disease) is primarily the result of inert gas bubbles; deafness and vertigo may result if the inner ear is involved. The most dramatic cause of disorientation under water is that due to vertigo. This vertigo is commonly a transient effect due to unequal caloric stimulation of the two labyrinths. The physical examination of the ear and nose necessary for assessment of diving fitness are discussed. A list of ENT contra-indications is presented which mandate temporary or permanent disqualification from diving. PMID- 3049475 TI - [Surgical interventions for ameliorating of equilibrium disorders refractory to conservative therapy]. AB - In cases of vertigo resistant to conservative treatment, surgery can be valuable for labyrinthine fistula, perilymphatic fistula, cupulolithiasis and Meniere's disease. For instance, covering a labyrinthine fistula by a fascial graft relieves the vertigo in an amazingly short time. Perilymphatic fistula is verified and treated by tympanotomy and is used for patients with special auditory and/or vestibular features. Cupulolithiasis usually requires no surgical treatment: special postural training will speed recovery, but a few cases need neurectomy of the inferior vestibular nerve. There are several methods of surgical treatment for Meniere's disease. In our experience neurectomy of the superior and inferior vestibular nerves on the diseased side together with decompression of the eighth cranial nerve via an extended transtemporal approach to the middle fossa is an excellent method of treating patients with vertigo attacks. PMID- 3049473 TI - [Principles of conservative therapy of peripheral and central disorders of equilibrium]. AB - Disturbance of equilibrium is a vestibular induced disturbance of orientation in space that is perceived subjectively as vertigo. Change of behaviour and specific medication is the optimal causal therapy, but symptomatic therapy is preferred in practice, using drugs that suppress different input activities or the activity of central vestibular structures. In this way a disordered flow of information is eliminated and the equilibrium is restored. PMID- 3049474 TI - [Rehabilitation of patients with vestibular disorders]. AB - The connections between the two vestibular nuclei permit vestibular compensation after unilateral lesions. The interactions between the vestibular and the visual and proprioceptive system can support vestibular compensation. Our physical training program for the treatment of vertigo is based on these mechanisms. The training consists of fixation exercises, smooth pursuit and motor learning by proprioceptive cognition. The results are very convincing. PMID- 3049477 TI - [Rehabilitation of patients with hearing disorders using hearing aids and tactical hearing measures]. AB - After a brief summary of the problems of rehabilitation of hearing-impaired subjects, the optimal conditions for rehabilitation with hearing aids are presented: a) The right timing is crucial. b) Optimal provision of hearing aids must be carried out in close co-operation between the patient, the hearing aid technician and the otolaryngologist. c) Easy handling of the hearing aid and the use of attachments must be guaranteed. d) The hearing-impaired person must be fully informed as to the extent and type of hearing loss. He/she must accept the affliction and know about the possibilities of rehabilitation. The patient's motivation is a pre-requisite for all further steps. e) The patients must learn tactical measures to make optimal use of their hearing ability in relation to their environment. Hearing tactics consist of hearing training and a change in the attitude of the hearing-impaired patients themselves and their attitude towards their surroundings. PMID- 3049476 TI - [Response latency characteristics for ENT medical assessment of auditory brain stem evoked response]. AB - A schematic description of the correlation between the various types of hearing disorders and the behaviour of auditory brain stem responses (ABR) is presented. Conductive pathology and high-frequency cochlear hearing loss prolong wave component latency due to energy loss and hair cell dysfunction. Latency is not affected in flat cochlear hearing loss. Prolonged interwave latencies between wave I and wave V indicate eighth nerve and brain stem disorders. An algorithm in the form of a flow chart was developed for location of the malfunction. Families of characteristics of wave V intensity-latency functions were designed for faster detection and more precise evaluation of conductive and cochlear hearing losses. PMID- 3049478 TI - [Malignant neoplasms of the lip, mouth, pharynx, nose, ear and larynx. A descriptive-epidemiologic study]. AB - Between 1952 and 1986 the mortality from malignant tumours of the head and neck in the German Federal Republic shows a varying picture: a decline of cancer of the lip, equivocal rates for cancer of the nose and ear, increases for cancer of the tongue and of the larynx, a marked increase for cancer of the floor of the mouth and of the hypopharynx, a lesser increase for cancer of the tonsillar area and other parts of the mouth, but almost no increase for cancer of the salivary glands. The trends are similar for males and females, but the male:female ratio increased during the observation period. The increase of mortality begins in the middle age group. Based on the data of the cancer registry of the Saarland it can be estimated that on average the incidence is twice as high as mortality. In general the trend is similar both in incidence and mortality (but not the degree of the trend). Multiple malignancies of the head and neck are observed more often than expected and patients should be followed up carefully. Combined clinical epidemiological studies are needed. PMID- 3049479 TI - [The status of hearing aid technology from the audiologic viewpoint]. AB - Although hearing aid technology has made considerable progress in the past, some important audiological demands are still unfulfilled. In particular, speech perception in noisy conditions must be improved. Traditional approaches to this fundamental problem, e.g. base cut, open mould fitting, horn equipped ear moulds, external receivers, in the ear hearing aids, directional microphones and audio input, are discussed, as well as modern concepts of automatic signal processing to improve signal-to-noise (S/N) ratio. The results of a study of speech discrimination with dual channel hearing aids with AGC in the low frequency channel reveal efficient noise reduction in specific S/N situations. However, in the USA self-adaptive noise filter systems in head-worn hearing aids, have proven to be inadequate using the present set of processing parameters. Multichannel and digital signal processing introduce additional unsolved problems into hearing aid fitting. PMID- 3049480 TI - Mitral valve prolapse--a pervasive condition of uncertain etiology. PMID- 3049481 TI - [A brief history of women]. PMID- 3049482 TI - Antecedents and consequences of variations in girls' maturational timing. AB - The antecedents and consequences of variations in girls' physical development are reviewed. Girls' development is highlighted because research on antecedents addresses genetic and environmental influences on menarcheal age variations, and because findings on the behavioral consequences of tempo variations have been less consistent for girls than for boys. Implications for adolescent health care are considered, particularly for the early maturing girl. PMID- 3049483 TI - Ultrasound examination of adolescent females with lower abdominal pain. AB - We evaluated pelvic ultrasound examination in adolescent females as an aid in the diagnosis of acute and chronic lower abdominal pain resulting from suspected gynecologic disease. Of 41 subjects, 35 (85.4%) had a final diagnosis of a gynecologic disorder. Pelvic ultrasound examination was positive in 19 of 35 (54.3%). Ten positive tests had relatively specific findings that supported (seven) or helped change (three) the initial clinical diagnosis. Nine positive tests had nonspecific findings that were consistent with (six) or helped change (three) the initial diagnosis. Twenty-one negative tests helped change the initial diagnosis (13); ruled out complications of acute salpingitis (five); or discriminated between alternative diagnoses (three). One test was falsely positive. Ultrasonography was most clearly cost-effective when surgery was being considered. We conclude that pelvic ultrasound examination may be a useful diagnostic adjunct in this type of adolescent patient. PMID- 3049484 TI - Effect of two types of hospital feeding gift packs on duration of breast-feeding among adolescent mothers. AB - Two hundred forty-four adolescent mothers under 18 years of age were surveyed during a 15-month period, and 53% elected to breast-feed. A subset of 60 primiparous breast-feeding adolescents were enrolled in an investigator-blind, randomized, prospective study to compare the effects on breast-feeding duration of a standard hospital discharge feeding gift pack containing formula and a specially designed study pack that was free of infant formula. Thirty-five percent of the 60 women breast-fed less than 1 month; 22% nursed longer than 1 month but less than 2 months; and 43% breast-fed more than 2 months. There was no significant difference in breast-feeding duration among mothers by gift pack group, although those who received the study gift pack rated it higher in usefulness (p less than (0.025). The provision of infant formula samples did not appear to have a deleterious effect on the duration of breast-feeding among a population of adolescent mothers. PMID- 3049486 TI - Teen mothering. Behaviors and interventions. AB - Data are reviewed that support the hypothesis that many adolescents interact with their infants in ways that may increase the infant's risk of developmental delay. The negative, long-term consequences of adolescent child-bearing create an environment that also augments this risk. Early intervention programs developed to address such risks are reviewed regarding their focus and content. Research designs used to evaluate their effectiveness are critiqued. The confounding of treatment approach with the frequency of contacts is a major limitation preventing an adequate evaluation of results. However, it is emphasized that despite a diversity of approaches to treatment and a lack of understanding as to what is responsible for the changes, intervention programs have been successful in improving adolescent maternal-infant interactions and/or enhancing infant development. PMID- 3049485 TI - The influence of anxiety and locus of control on adolescents' response to naproxen sodium for mild to moderate pain. AB - This study assessed the influence of internal health locus of control (IHLC) and anxiety on the adolescent's response to the treatment of mild and moderate pain. Fifty-four adolescents (ages 16-22 years) from two adolescent clinics presenting with mild to moderate pain caused by dysmenorrhea, muscle sprain or strain, headache, or backache were studied. Following a physical examination and a pretest assessment of pain, IHLC, and the Spielberger State-Trait Anxiety Inventory, subjects were randomly assigned in a double-blind fashion to groups receiving placebo (n = 16), 100 mg of naproxen sodium (n = 19), or 200 mg naproxen sodium (n = 19) and assessed at 1, 2, 3, and 4 hours. Based on a repeated-measure analysis of covariance test, there were no differences between groups in the pretest measurements. All treatment groups had a decrease in pain over the 4 hours (p less than 0.0001). Patients from one institution had more pain reduction than at the other (p less than 0.0001), and females had more pain reduction than males (p less than 0.034). Subjects receiving 200 mg of naproxen sodium had more pain relief (p less than 0.034) than subjects taking placebo at hour 2. Baseline anxiety was positively associated with pain after receiving placebo, but inversely associated with pain after taking naproxen sodium. The IHLC appeared to have a positive effect on the response to naproxen sodium, but no effect on the response to placebo. PMID- 3049487 TI - Distracted cervical spinal fusion for management of caudal cervical spondylomyelopathy in large-breed dogs. AB - Using an autogenous bone graft (obtained from the iliac crest), 4-mm cancellous bone screws, and polymethylmethacrylate, a distracted cervical spinal fusion technique was performed on 10 dogs with myelographic evidence of caudal cervical spondylomyelopathy. All dogs had evidence of dynamic soft tissue spinal cord compression, as indicated by flexion, extension, and traction myelographic views. Of the 10 dogs, 4 previously had undergone surgery by use of ventral slot or cervical disk fenestration techniques, and their neurologic status had deteriorated after the original surgery. Preoperative neurologic status of the 10 dogs included nonambulatory tetraparesis (n = 5), severe ataxia with conscious proprioceptive deficits (n = 2), and mild ambulatory ataxia with conscious proprioceptive deficits (n = 3). Five dogs had signs of various degrees of cervical pain. Clinical improvement was observed in 8 of 10 dogs--either improved neurologic status or elimination of cervical pain. Implant loosening developed in 3 dogs; 2 of them were euthanatized because of lack of neurologic improvement. Radiographic evidence of bony cervical fusion was observed during a 9- to 24-week period in 6 of the 8 surviving dogs. The distracted cervical fusion technique appears to be a valid surgical procedure to manage cervical spondylomyelopathy in those dogs in which the lesions are limited to one cervical intervertebral disk space. PMID- 3049488 TI - People and events influence life. PMID- 3049490 TI - Treatment of respiratory distress in a prematurely born foal. AB - A foal born 3 weeks prematurely was treated for respiratory distress, using a combination of oxygen therapy and mechanical ventilatory assistance. Clinical response and arterial blood gas tensions were monitored regularly. Continuous positive-airway pressure and intermittent positive-pressure ventilation administered via a nasotracheal tube were effective in improving arterial oxygenation and ventilatory function. PMID- 3049489 TI - Bar suture (toggle pin) vs open surgical abomasopexy for treatment of left displaced abomasum in dairy cattle. AB - Fifty-nine dairy cows with left displacement of the abomasum were randomly assigned to receive 1 of 2 treatments. Twenty-eight were treated by right paramedian abomasopexy via laparotomy, and 31 were treated by percutaneous fixation of the abomasum, using the bar suture or toggle pin technique. Comparisons of the 2 groups, at 4 follow-up examinations, yielded no significant differences in return to normal milk production, return to normal feed intake, mortality, culling rate, tissue reaction at the surgical site, or redisplacement up to 60 days into the subsequent lactation. PMID- 3049491 TI - L-656,575 (OCP-9-176): a novel oxacephem. Pharmacokinetics and experimental chemotherapy. AB - L-656,575 is a new oxacephem that, based on studies in rhesus monkeys, is expected to have a moderately long half-life in humans. After administration of a 10-mg/kg dose by the intramuscular route to rhesus monkeys, peak serum concentrations of 32-54 micrograms/ml were seen at about 30 minutes, and the half life was estimated to be 63 minutes. Urinary recovery of administered dose was greater than 94% in 6 hours. In mice given a 20-mg/kg dose by the subcutaneous route, a peak serum concentration of 22.9 microgram/ml was observed at 15 minutes after dosing, and the half-life in serum was about 18 minutes. Urinary recovery of the dose was 59% in 6 hours. In another study in mice, administration of probenecid did not extend the half-life of L-656,575, suggesting that the antibiotic is excreted primarily by glomerular filtration in this species. Binding to human plasma proteins was 30% at drug concentrations from 25-100 micrograms/ml. L-656,575 also was shown to be efficacious in experimental bacteremias due to Gram-positive and Gram-negative pathogens in mice, thus confirming the broad spectrum of activity demonstrated for L-656,575 in vitro. PMID- 3049492 TI - Comparative anatomy of the cochlea and auditory nerve in mammals. AB - The numbers and structure of hair cells; afferent, efferent, and reciprocal synapses as seen at the base of hair cells; innervation patterns of first order cochlear neurons; and number and morphology of spiral ganglion cells will be discussed and compared in the guinea pig, rat, cat, monkey and man. Despite many similarities both in the organ of Corti and the spiral ganglion in these species, there are a number of differences which may have important physiologic implications. In the organ of Corti, the major differences among species are the length and width of the basilar membrane, the number of inner and outer hair cells, and the length of hairs on both inner and outer hair cells. Significant differences in the innervation pattern of the inner hair cell among these species include the number of afferent nerve terminals per inner hair cell, the degree of branching of afferent fibers, and the number of synapses per afferent nerve terminal. Among outer hair cells, the number of afferent nerve terminals per outer hair cell, presence or absence of a pre-synaptic body, presence or absence of reciprocal synapses, the number of efferent terminals per outer hair cell, and the presence of dendodendritic synapses in outer spiral bundles may be differences important physiologically. In the spiral ganglion, there are significant differences in the number of spiral ganglion cells, the number of cochlear nerve fibers, the percentage of spiral ganglion cells which are myelinated, and the presence of synapses on spiral ganglion cells. PMID- 3049493 TI - Physiological and psychophysical correlates of temporal processes in hearing. AB - Discharges of auditory nerve fibers are synchronized to stimulus frequencies below 4-5 kHz. The phase-locking phenomenon has been studied in considerable detail in several animal species. Although strikingly close correspondences exist between phase-locking behavior in animals and human perceptual performance on certain tasks, there is still no clear evidence that the human brain actually bases perceptual decisions on temporally encoded frequency information. The alternative to temporal coding is rate-place coding, in which frequency is assigned on the basis of peaks in cochlear excitation patterns. This paper reviews pertinent physiological, psychophysical and modeling data in three classes of experiment whose results are explanable in terms of both rate-place and temporal processing of neural responses. The experiments deal with the pitch of complex tones, vowel identification, and pure-tone frequency discrimination. The data described here suggest that temporal models of frequency coding compete well with and in some cases offer a more parsimonious explanation of perceptual performance than rate-place codes do, particularly at low and middle frequencies. A potentially important implication of the analyses conducted here is that humans may not code frequency information in synchronized activity as well as other species. The data suggest that within limits the human ear is capable of using either temporal and rate-place frequency codes, and that the specific code employed by the perceptual processor is task-dependent. PMID- 3049494 TI - Plant-animal interactions in larkspur poisoning in cattle. AB - Larkspur (Delphinium spp.) toxicity in cattle seriously impedes the efficient use of productive mountain rangelands. Larkspurs contain complex diterpenoid alkaloids that cause acute intoxication and death from respiratory paralysis. Alkaloids and their concentrations vary among larkspur species, plant parts and phenological growth stages, thus causing great variability in toxicity. Ingestion rate of larkspur by the cow, alkaloid toxicity and concentration in the plant and the kinetics of absorption and excretion interact to determine whether a cow is poisoned. Plant and animal factors influencing consumption and subsequent intoxication must be further elucidated to devise management strategies to reduce liverstock losses. PMID- 3049495 TI - Toxicity and metabolism of pyrrolizidine alkaloids. AB - Pyrrolizidine alkaloids (PA) are found mainly in plants of three families: boraginaceae, Compositae and Leguminosae. In North America, PA poisoning of livestock is caused primarily by consumption of Senecio and Crotalaria spp. The PA of Senecio spp. cause irreversible hepatic damage; toxicity signs are a consequence of impaired liver function. Crotalaria intoxication leads to pulmonary damage as a primary effect; hepatic effects are less prominent. Large species differences exist in susceptibility to PA toxicosis. Small herbivores such as sheep, goats, rabbits, guinea pigs and other herbivorous laboratory animals are highly resistant to PA toxicity, associated with a low rate of hepatic production of reactive metabolites (pyrroles) and(or) a high rate of activity of detoxifying enzymes. Diester PA common to Heliotropium and Echium spp. are metabolized in the ovine rumen to 1-methyl metabolites, whereas the macrocyclic ester PA of Senecio spp. are not. Exposure to PA results in high concentrations of liver Cu, reduced liver Zn, and abnormal Fe metabolism with hematopoiesis markedly impaired. Pyrrolizidine alkaloid toxicity alters vitamin A metabolism in rats, depressing plasma and liver levels of vitamin A. Synthetic antioxidants in the diet confer protective activity in laboratory animals (e.g., rats, mice) against PA toxicoses. The PA and their metabolites are secreted in the milk of lactating animals, but this probably does not represent a significant human health hazard. PMID- 3049496 TI - Researching the plant-animal interface: the investigation of ingestive behavior in grazing animals. AB - Profitable livestock production from forages largely depends on efficiency of converting forages into products. Efficient grazing management systems require an understanding of the roles of system components. However, experimentation should be conducted with regard to the system as a whole rather than on the systems components in isolation. This may necessitate development of computer models. The short-term intake of forage by grazing animals is controlled both by the structure of the forage and by effects of the ingested forage on gut fill as moderated by the hunger-satiety complex. Intake can be defined as the product of bite size, rate of biting and grazing time. Measurement of these variables is facilitated by the use of esophageally fistulated animals and automatic recording devices. Bite size has the greatest influence on intake, with rate of biting and grazing time being compensatory variables. Sward structure influences bite size to varying degrees. In temperate grass swards, leaf surface height appears to be the dominant influence on bite size. But in tropical grass swards, leaf density and leaf:stem ratio have a greater influence on bite size than does leaf surface height. Alternative techniques to conventional grazing trials are described. Diversity of environments and forages in the U.S. requires further research into the development of grazing systems. In the future, small-scale trials and computer simulation techniques likely will be used to a greater extent. PMID- 3049497 TI - Toxicoses in livestock from the hemlocks (Conium and Cicuta spp.). AB - The hemlocks, Conium maculatum (poison-hemlock) and Cicuta spp. (waterhemlock), are poisonous plants that cause sizeable losss to the livestock industry. Clinical signs of poisonhemlock toxicosis are similar in all species of livestock and include muscular weakness, incordination, trembling, initial central nervous system stimulation, depression and death from respiratory paralysis. Poison hemlock also causes skeletal defects in the offspring of cattle, pigs and sheep and cleft palate in pigs when ingested during specific periods of gestation. The primary toxicants in poison-hemlock are coniine and gamma-coniceine. Coniine predominates in mature plants and seed, whereas gamma-coniceine predominates in early growth of the plant. Waterhemlock is the most violently toxic poisonous plant known. The toxicant is cicutoxin, which acts on the central nervous system, causing violent convulsions and death. Clinical signs of poisoning appear within 15 min after ingestion of a lethal dose and include excessive salivation, nervousness, tremors, muscular weakness and convulsive seizures interspersed by intermittent periods of relaxation and a final paralytic seizure resulting in anoxia and death. Elevated activities of lactic dehydrogenase, aspartate aminotransferase and creatine kinase in blood are observed, indicative of muscular damage. Toxicoses from poisonhemlock and waterhemlock generally occur in early spring when both plants emerge before other, more palatable plants begin to grow. All parts of the poison-hemlock plant are toxic. The root or tubers of waterhemlock are toxic; however, experimental evidence concerning the toxicity of other plant parts is inconclusive. PMID- 3049498 TI - Livestock models of human birth defects, reviewed in relation to poisonous plants. AB - Certain birth defects in livestock induced by poisonous plants possess clinical similarities to congenital deformities in humans. They meet some criteria as models for those corresponding human conditions. They are induced by several plants, including members of the Astragalus, Lupinus, Conium, Nicotiana and Veratrum genera. The terata expressions include effects in bone and soft tissue, particularly in the limbs, spinal cord and cephalic regions. Whether these livestock conditions become extensively used as models of human terata will be determined by how well they meet the criteria sought in good models. These prospective models have many favorable characteristics; however, being large domestic livestock, they have certain logistic disadvantages compared with small rodents as models. PMID- 3049499 TI - Evaluation of a monoclonal antibody-based immunoassay for detecting type B Clostridium botulinum toxin produced in pure culture and an inoculated model cured meat system. AB - A monoclonal antibody-based amplified ELISA method for detecting Clostridium botulinum type B toxin was evaluated for its ability to detect the toxin in the supernatant fluid of pure cultures and after growth from Cl. botulinum spores inoculated into pork slurries. Slurries containing NaCl (1.5-4.5% w/v) and polyphosphate (0.3% w/v) were either unheated or heated 80 degrees C/5 min followed by 70 degrees C/2 h before incubation at 15 degrees, 20 degrees or 27 degrees C. Presence of specific toxin was confirmed by mouse bioassay and results were compared with those of the amplified ELISA method. A total of 48 strains, consisting of 38 Cl. botulinum and 10 Cl. sporogenes (putrefactive anaerobes), and 140 slurry samples were tested. Cultures of eight out of nine strains of type B Cl botulinum and 73 of 101 slurry samples containing type B toxin were positive by ELISA; the remaining 28 slurry samples contained type B toxin at levels below or close to the detection limit (20 LD50/ml) of the type B ELISA. No false positive reactions occurred with Cl. botulinum types A, C, D, E or F, or with the 10 strains of Cl. sporogenes. Toxin produced by one strain of Cl. botulinum type B (NCTC 3807) was not detected by this single monoclonal antibody-based amplified ELISA. With a mixture of two monoclonal antibodies, however, the toxin from NCTC 3807 could be detected without reducing the sensitivity of the ELISA. PMID- 3049501 TI - An improved procedure for shipboard enumeration of faecal indicator bacteria in marine sediments from sewage sludge disposal areas. AB - An improved membrane filtration procedure for use on board ship to enumerate Escherichia coli and Group D faecal streptococci in marine sediments is described. Ultrasonication extraction combined with resuscitation of sublethally injured cells yielded significantly higher counts of E. coli than sediments shaken by hand. Counts of E. coli were also higher on mFC agar (without rosalic acid) after a period of resuscitation on tryptone-soy agar supplemented with 0.1% yeast extract than on a 4% Teepol-lactose medium. Ultrasonication of sediments made no significant difference to counts of Group D faecal streptococci on KF streptococcus agar. These improved isolation procedures allowed better discrimination of the area affected by sewage sludge at a disposal site off the northeast coast of England. PMID- 3049500 TI - Monitoring for the development of antimicrobial resistance during the use of olaquindox as a feed additive on commercial pig farms. AB - Since 1982, when olaquindox was introduced as a pig-feed additive in the UK, about 12 commercial farms in Suffolk have been monitored annually to check for the possible emergence of resistance to olaquindox and chloramphenicol among the coliform flora of the pigs and their environment. In spite of the sampling variability and the impossibility of controlling the use of feed additives and management on the farms, the overall results obtained were consistent and, it is suggested, the method is widely applicable. A steady, albeit low, increasing incidence and level of resistance to olaquindox was recorded (1982-1984) on farms using it and, to a lesser degree, on neighbouring farms that did not. No significant increase in the level of chloramphenicol resistance was observed. Genetical studies on a selection of olaquindox-resistant isolates suggested that the genes determining resistance were likely to be borne on the chromosome. PMID- 3049502 TI - Antibiotic resistance among coliforms and Pseudomonas spp. from bodies of water around Port Harcourt, Nigeria. AB - Samples from municipal waste water, the Bonny River estuary and wells in and around Port Harcourt were examined for bacteriological quality over a 9 month period. A total of 157 Pseudomonas spp., 133 Escherichia coli and 282 other coliforms were isolated and tested for the incidence of resistance to 10 antibiotics. All of the Pseudomonas spp. were resistant to at least one of the antibiotics while 96.2% were resistant to two or more. Most (83.5%) of the E. coli and other coliforms (91.8%) were resistant to at least one antibiotic. All strains were susceptible to gentamicin. Minimal inhibitory concentrations of ampicillin and tetracycline for E. coli ranged from 6.25 to 50 and 6.25 to 12.5 micrograms/ml, respectively. Minimal inhibitory concentrations of ampicillin and tetracycline were 1000 and 25 micrograms/ml for the Pseudomonas strains. The high incidence of bacterial resistance to antibiotics is discussed in relation to the widespread use of antibiotics, and possible public health implications. PMID- 3049503 TI - John Geoffrey Carr 1927-1987. PMID- 3049504 TI - Detection of salmonellas in confectionery products by conductance. AB - A modified lysine decarboxylase broth has been developed which could be used with a Bactometer M123 to differentiate salmonellas from other bacteria by the characteristics of the conductance detection curve. The medium was used in combination with a selenite cystine trimethylamine oxide dulcitol medium to screen 50 strains of salmonellas and 42 strains of other organisms to establish detection curve magnitude and rate values which could be used to identify curves specific to salmonellas. The combination of media detected all salmonellas tested except Salmonella pullorum. The two media were used to screen 100 inoculated product samples with the Bactometer instrument, in parallel with traditional plating procedures, and using various combinations of pre-enrichment and selective enrichment incubation periods. After 24 h pre-enrichment, the Bactometer system detected more positive samples than the conventional plating procedures after pre-enrichment and selective enrichment. It is considered that these media used in parallel in the Bactometer after conventional pre-enrichment could provide a 48 h screening procedure for salmonellas with a sensitivity comparable to present plating procedures. PMID- 3049505 TI - Olaquindox resistance in the coliform flora of pigs and their environment: an ecological study. AB - Faecal samples were taken weekly from a chosen pen on each of four commercial pig farms, two of which used olaquindox as a feed additive. Coliform bacteria were isolated from the samples and the incidence and level of resistance to olaquindox and to four therapeutic antibiotics determined. The coliforms isolated were biotyped to follow the emergence of drug-resistant sub-populations. The results showed that the coliform flora was complex and that a turnover of biotypes was associated with changes in the occupancy of the pen and, possibly, diet. This turnover led to large fluctuations in both incidence and level of resistance to olaquindox and the antibiotics. Olaquindox resistance was not specifically linked with resistance to any therapeutic antibiotic, and the possible origin of olaquindox-resistant strains is discussed in relation to the biotypes. PMID- 3049506 TI - Inactivation of bacteria by Purogene. AB - The bacteriocidal efficacy of Purogene, a stabilized aqueous solution of chlorine dioxide (ClO2) was examined using bacteria of concern to public health. The organisms tested were: Escherichia coli, Pseudomonas aeruginosa, Yersinia enterocolitica, Klebsiella pneumoniae, Streptococcus pyogenes Group A, Salmonella typhimurium and Bacillus subtilis. The test organisms responded differently to inactivation by Purogene. At least a 4 log reduction in bacterial counts was noted when Purogene was applied at a concentration of 0.75 mg/l. Since Purogene is a stabilized complex, it was necessary to provide a chemical environment suitable for the release of ClO2 in this solution. This was done by varying the pH of Purogene from 3.5 to 8.6 (pH of Purogene is 8.6) while keeping the pH of the experimental medium constant (pH 7.0). The results showed that Purogene was most efficacious at the lowest pH tested (pH 3.5). This indicates that as chlorine dioxide solutions were reduced to chlorite (which predominates at pH 8.6), their bacteriocidal efficacy was reduced, suggesting free chlorine dioxide as the active disinfecting species. PMID- 3049508 TI - Starvation and nutrient resuscitation of Klebsiella pneumoniae isolated from oil well waters. AB - Klebsiella pneumoniae isolated from oil well waters reduced in size in response to nutrient starvation. The cells remained viable during starvation and later were able to grow rapidly when stimulated by nutrients. The heterotrophic potential, culture absorbance and extracellular polysaccharide production decreased during cell starvation whereas an initial increase in colony-forming units was observed on agar plates. Transmission electron microscopy (TEM) after 24 d revealed that the cells had changed to small rods or cocci between 0.5 by 0.25 micron and 0.87 by 0.55 micron. When transferred to half-strength brain heart infusion medium, TEM showed cell division and rod-shaped cells after 45 min and full resuscitation within 4 h. Cell response was much slower in sodium citrate medium and resuscitation took 8 h. PMID- 3049507 TI - Production of antibody to lipopolysaccharide (LPS) after immunization with a LPS polymyxin B-agarose immunogen. AB - A method was devised to produce antibodies to lipopolysaccharide (LPS) in guinea pigs following a single immunization. The antigen was prepared by mixing polymyxin B-agarose with LPS from Escherichia coli O55:B5. Use of the agarose support allowed purification of the complex by simple washing procedures. Twenty nine days after a single injection of the immunogen mixed with Freund complete adjuvant all animals demonstrated antibody to the LPS portion of the complex. No antibodies were detected to the polymyxin B component. Typical titres of LPS as measured by ELISA were 2(11). After, a booster immunization, titres of LPS antibody were further increased and a greater avidity was noted. In contrast to other methods which have been employed for production of antibody to LPS, use of the polymyxin B-agarose complex has the following advantages: ease of antigen preparation, ready purification of the complex, potent immunostimulation, and under the conditions employed here, LPS-specific antibody production, without accompanying antibody to polymyxin B. PMID- 3049509 TI - Novel plasmid-mediated ceftazidimase from Klebsiella pneumoniae isolates. PMID- 3049510 TI - Hydrolysis of semi-synthetic macrolides by erythromycin esterase from Escherichia coli. PMID- 3049511 TI - Regulation of lactic acid production during exercise. AB - Lactic acid accumulates in contracting muscle and blood beginning at approximately 50-70% of the maximal O2 uptake, well before the aerobic capacity is fully utilized. The classical explanation has been that part of the muscle is O2 deficient and therefore lactate production is increased to provide supplementary anaerobically derived energy. Currently, however, the predominant view is that lactate production during submaximal dynamic exercise is not O2 dependent. In the present review, data and arguments in support of and against the hypothesis of O2 dependency have been scrutinized. Data underlying the conclusion that lactate production during exercise is not O2 dependent were found to be 1) questionable, or 2) interpretable in an alternative manner. Experiments in human and animal muscles under various conditions demonstrated that the redox state of the muscle is reduced (i.e., NADH is increased) either before or in parallel with increases in muscle lactate. Based on experimental data and theoretical considerations, it is concluded that lactate production during submaximal exercise is O2 dependent. The amount of energy provided through the anaerobic processes during steady-state submaximal exercise is, however, low, and the role of lactate formation as an energy source is of minor importance. It is proposed that the achievement of increased aerobic energy formation under conditions of limiting O2 availability requires increases of ADP, Pi, and NADH and that the increases in ADP (and therefore AMP via the adenylate kinase equilibrium) and Pi will stimulate glycolysis, and the resulting increase in cytosolic NADH will shift the lactate dehydrogenase equilibrium toward increased lactate production. PMID- 3049512 TI - Stimulatory role for endogenous opioid peptides on postexercise insulin secretion in rats. AB - Endogenous opioid peptides (EOP) and prior exercise may modulate the stimulatory effect of glucose on insulin secretion. To gain insights into these relationships, we studied male Wistar rats (187-245 g) during sustained hyperglycemia by use of the glucose clamp technique. Four groups of sedentary fed rats (n = 8/group) either ran (Ex) at 24 m/min, 0% grade, or rested (R) for 40 min. Thirty minutes after Ex or R, arterial blood glucose was elevated to and maintained at 11 mM for 2 h by a variable glucose infusion. At the start of Ex or R rats had saline (Sal) or naloxone (Nal, an opioid antagonist) intravenous infusions for 160 min (40 min Ex + 30 min R + 90 min of a 120-min glucose clamp). Steady-state glucose infusion rates (SSGIR) were approximately 55 mg.kg-1.min-1 at the start of the clamp and declined significantly over the 2nd h to approximately 45 mg.kg-1.min-1. No significant differences existed in SSGIR between groups. R-Sal and Ex-Sal groups did not differ in their insulin response to hyperglycemia. In contrast, when all groups were compared at the end of the Nal or Sal infusion, Ex-Nal had the lowest insulin concentration (749 +/- 174 pmol/l), whereas the R-Nal group had the highest (1,581 +/- 216 pmol/l, P less than 0.05). These data suggest a stimulatory role for EOP on insulin secretion that is expressed after a prior stress (Ex). Thus one function of exercise induced activation of EOP may be to regulate insulin secretion in the immediate postexercise period. PMID- 3049513 TI - Tracheal mucosal blood flow responses to autonomic agonists. AB - In lightly anesthetized adult sheep, we determined tracheal mucosal blood flow (Qtr) by measuring the steady-state uptake of dimethyl ether from a tracheal chamber created by an endotracheal tube provided with two cuffs. Qtr normalized for carotid arterial pressure [Qtr(n)] was determined before and after the exposure of the tracheal mucosa to aerosolized phenylephrine (0.25-2.0 mg), isoproterenol (0.05-0.8 mg), and methacholine (2.5-20 mg). The same doses of methacholine were also administered during the intravenous infusion of vasopressin. The measurements were repeated after intravenous pretreatment with the respective antagonists phentolamine, propranolol, and atropine. Mean +/- SE base-line Qtr(n) was 1.2 +/- 0.1 ml.min-1.mmHg-1.10(2). The autonomic antagonists had no effect on mean Qtr(n). Phenylephrine produced a dose-dependent decrease in mean Qtr(n) (-70% at the highest dose), which was blunted by phentolamine, and isoproterenol produced a dose-dependent increase in mean Qtr(n) (40% at the highest dose), which was blocked by propranolol. Methacholine failed to alter mean Qtr(n) even when Qtr was first decreased by vasopressin. We conclude that in lightly anesthetized adult sheep 1) base-line Qtr(n) is not under adrenergic or cholinergic control, 2) a locally administered alpha-adrenergic agonist decreases and beta-adrenergic agonist increases Qtr(n) via specific receptor activation, and 3) a locally administered cholinergic muscarinic agonist has no effect on Qtr(n). PMID- 3049514 TI - Effect of exercise training on glucose homeostasis in normal and insulin deficient diabetic rats. AB - The effect of 8-wk of treadmill training on plasma glucose, insulin, and lipid concentrations, oral glucose tolerance, and glucose uptake in the perfused hindquarter of normal and streptozocin-treated, diabetic Sprague-Dawley rats was studied. Diabetic rats with initial plasma glucose concentrations of 200-450 mg/dl and control rats were divided into trained and sedentary subgroups. Training resulted in lower plasma free fatty acid concentrations and increased triceps muscle citrate synthase activity in both the control and diabetic rats; triglyceride concentrations were lowered by training only in the diabetic animals. Oral glucose tolerance and both basal and insulin-stimulated glucose uptake in hindquarter skeletal muscle were impaired in the diabetic rats, and plasma glucose concentrations (measured weekly) gradually increased during the experiment. Training did not improve the hyperglycemia, impaired glucose tolerance, or decreased skeletal muscle glucose uptake in the diabetic rats, nor did it alter these parameters in the normal control animals. In considering our results and those of previous studies in diabetic rats, we propose that exercise training may improve glucose homeostasis in animals with milder degrees of diabetes but fails to cause improvement in the more severely insulin-deficient, diabetic rat. PMID- 3049515 TI - Glucose transport into rat skeletal muscle: interaction between exercise and insulin. AB - This study was done to evaluate the effect of insulin on sugar transport into skeletal muscle after exercise. The permeability of rat epitrochlearis muscle to 3-O-methylglucose (3-MG) was measured after exposure to a range of insulin concentrations 30, 60, and 180 min after a bout of exercise. Thirty and 60 min after exercise, the effects of exercise and insulin on 3-MG transport were additive over a wide range of insulin concentrations, with no increase in sensitivity or responsiveness to insulin. After 180 min, when approximately 66% of the exercise-induced increase in sugar transport had worn off, both the responsiveness and sensitivity of the glucose transport process to insulin were increased. These findings appear compatible with the hypothesis that the actions of exercise and insulin result in activation and/or translocation into the plasma membrane of two separate pools of glucose transporters in mammalian skeletal muscle. PMID- 3049516 TI - Stimulation of ciliary beat frequency by autonomic agonists: in vivo. AB - beta 2-Adrenergic bronchodilator and muscarinic cholinergic bronchoconstrictor agonists both stimulate ciliary activity in vitro. To test the hypothesis that increases in autonomic activity would result in increases in ciliary beat frequency (CBF) in vivo, a correlation analysis heterodyne laser light-scattering system was developed and validated to measure the stimulating effects of sympathomimetic and parasympathomimetic agonists on tracheal CBF in intact, anesthetized beagles. The mean baseline CBF from 42 studies of 274 measurements in 9 (5 male and 4 female) adult beagles was 6.6 +/- 1.1 Hz. The stimulating effects of a beta 2-adrenergic agonist, fenoterol, and a muscarinic cholinergic agonist, methacholine, on CBF were studied on four and eight beagles, respectively. The studies were randomized and blinded. Aerosolized 10(-5) M fenoterol stimulated the CBF from the base line of 6.8 +/- 2.5 to 32.0 +/- 17.9 Hz in four dogs. Aerosolized methacholine stimulated the CBF from the base line of 5.8 +/- 0.7 to 9.4 +/- 3.0 Hz for 10(-8) M, and to 12.6 +/- 3.1 Hz for 10(-6) M in eight dogs. These are the first data obtained in intact animals that demonstrate CBF in the lower respiratory tract is regulated by autonomic agonists. PMID- 3049517 TI - An improved method for primary culture of ovarian androgen-producing cells in serum-free medium: effect of lipoproteins, insulin, and insulinlike growth factor I. AB - Although luteinizing hormone (LH) alone stimulates ovarian interstitial cells cultured in serum-free medium to synthesize large amounts of androgens, there seem to be additional factors in vivo that modulate the time course and magnitude of the cellular responses to LH. In an attempt to develop a more nearly physiologic cell culture model, lipoproteins, insulin, and insulinlike growth factor-I (IGF-I) were added to the serum-free medium. The effects of these modifications on androgen biosynthesis by dispersed cells from ovaries of hypophysectomized immature rats cultured in 96-well tissue culture plates were examined. A saturating dose of LH stimulated a 25-fold increase in androsterone synthesis at 2 d, which decreased at 4 and 6 d. Addition of human high density (hHDL) or human low density lipoprotein (hLDL) caused a 2.5-fold increase in LH stimulated androsterone synthesis. Cells were approximately twice as sensitive to hHDL (ED50 = 5.5 +/- 0.5 micrograms cholesterol/ml) compared to hLDL (ED50 = 9.1 +/- 1.1 micrograms cholesterol/ml). Surprisingly, rat HDL caused only a 40% increase in LH-stimulated androsterone synthesis. When insulin alone was added to cells cultured with a saturating dose of LH, there was a 2.8-fold increase in androsterone synthesis. Addition of hHDL and insulin together caused a synergistic increase in LH-stimulated androsterone synthesis. In contrast to hHDL, which did not change the time course of LH-stimulated androsterone production, insulin prolonged maximal LH-stimulated androsterone synthesis at 4 and 6 d. Inasmuch as the ED50 for insulin action (1.3 +/- 0.1 micrograms/ml) was supraphysiologic, the effects of IGF-I on LH-stimulated androgen synthesis were examined. IGF-I mimicked all of the effects of insulin, but at a physiologic concentration (ED50 = 2.5 +/- 0.3 ng/ml). Ovarian cells cultured in serum-free medium supplemented with hHDL and insulin or IGF-I exhibit responses that closely approximate the physiologic responses observed in vivo. These results suggest that lipoproteins and IGF-I are important physiologic stimulators of ovarian theca-interstitial cell androgen biosynthesis which, when added to the serum-free medium, make the cellular responses in this in vitro model more nearly approximate the responses in vivo. PMID- 3049518 TI - A new serum-free method of measuring growth factor activities for human breast cancer cells in culture. AB - Growth of the MCF-7, T47D, and ZR-75-1 human breast cancer cells was established in a serum-free defined medium (MOM-1) composed of a 1:1 (vol/vol) mixture of Ham's F12 medium and Dulbecco's modified Eagle's medium containing 15 mM HEPES (pH 7.2), 2 mM 1-glutamine, 20 micrograms/ml glutathione, 10 micrograms/ml insulin, 10 micrograms/ml transferrin (Tf), 10 ng/ml selenous acid, 0.3 nM triiodothyronine, 50 micrograms/ml ethanolamine, 20 ng/ml epidermal growth factor, 2.0 nM 17 beta-estradiol, and 1.0 mg/ml bovine serum albumin (BSA). Proliferation in MOM-1 was 50 to 70% of the serum stimulated rate. Deletion of components from MOM-1 gave a medium (Tf-BSA) containing only HEPES, 10 micrograms/ml Tf, and 200 micrograms/ml BSA, which sustained MCF-7 and T47D cells in a slowly dividing and mitogen responsive state; ZR-75-1 cells required Tf plus 1.0 mg/ml BSA. In Tf-BSA, insulin and insulin-like growth factor I (IGF-I) were mitogenic with ED50 values of 2 to 3 ng/ml and 30 to 150 pg/ml, respectively, with MCF-7 cells. The T47D cells were responsive to these factors in Tf-BSA but required 10-fold higher concentrations for ED50. At saturating concentrations, insulin and IGF-I promoted 1.5 to 3.5 cell population doublings over controls in 8 d. At less than or equal to ng/ml concentrations, epidermal growth factor, insulin-like growth factor II, and basic fibroblast growth factor were mitogenic for human breast cancer cells in Tf-BSA. Mitogen activities in uterus and pituitary extracts were assayed readily in Tf-BSA. This new method offers a convenient means of comparing the potencies of growth-promoting factors on human breast cancer cells without interfering activities known to be present in serum. PMID- 3049520 TI - [Magnetic resonance imaging in senology]. PMID- 3049521 TI - Magnetic resonance imaging of focal hepatic lesions: a review. PMID- 3049519 TI - Social and pragmatic deficits in autism: cognitive or affective? AB - Autism is characterized by a chronic, severe impairment in social relations. Recent studies of language in autism also show pervasive deficits in pragmatics. We assume, uncontroversially, that these two deficits are linked, since pragmatics is part of social competence. This paper reviews the literature describing these deficits, and then considers two different psychological theories of these phenomena: the Affective theory and the Cognitive theory. Although the Affective theory makes better sense of the results from emotional recognition tasks, the Cognitive theory predicts the particular pattern of impaired and unimpaired social skills in autism, as well as the pragmatic deficits. These two theories might usefully be integrated in the future. PMID- 3049522 TI - [Magnetic resonance imaging in the study of renal transplants: value and limits]. PMID- 3049523 TI - Magnetic resonance imaging of bones and soft tissues. A review of the current literature. PMID- 3049524 TI - [Echography in the infertility clinic]. PMID- 3049525 TI - Acute suppurative thyroiditis in a child. PMID- 3049527 TI - Cervical cancer. Epidemiology, histopathology, diagnosis. PMID- 3049526 TI - [Current imaging technics and interventional radiology in thoracic pathology. Viewpoint of the pneumologist]. PMID- 3049529 TI - Treatment of uterine cervix cancer. PMID- 3049528 TI - Diagnostic evaluation in carcinoma of the uterine cervix: imaging methods. PMID- 3049530 TI - Cloning and expression in Escherichia coli of the Alcaligenes eutrophus H16 poly beta-hydroxybutyrate biosynthetic pathway. AB - The poly-beta-hydroxybutyrate (PHB) biosynthetic pathway from Alcaligenes eutrophus H16 has been cloned and expressed in Escherichia coli. Initially, an A. eutrophus H16 genomic library was constructed by using cosmid pVK102, and cosmid clones that encoded the PHB biosynthetic pathway were sought by assaying for the first enzyme of the pathway, beta-ketothiolase. Six enzyme-positive clones were identified. Three of these clones manifested acetoacetyl coenzyme A reductase activity, the second enzyme of the biosynthetic pathway, and accumulated PHB. PHB was produced in the cosmid clones at approximately 50% of the level found in A. eutrophus. One cosmid clone was subjected to subcloning experiments, and the PHB biosynthetic pathway was isolated on a 5.2-kilobase KpnI-EcoRI fragment. This fragment, when cloned into small multicopy vectors, can direct the synthesis of PHB in E. coli to levels approaching 80% of the bacterial cell dry weight. PMID- 3049532 TI - Two regions of mature periplasmic maltose-binding protein of Escherichia coli involved in secretion. AB - Six mutations in malE, the structural gene for the periplasmic maltose-binding protein (MBP) from Escherichia coli, prevent growth on maltose as a carbon source, as well as release of the mutant proteins by the cold osmotic-shock procedure. These mutations correspond to insertion of an oligonucleotide linker, concomitant with a deletion. One of the mutations (malE127) affects the N terminal extension (the signal peptide), whereas the five others lie within the mature protein. As expected, the export of protein MalE127 is blocked at an early stage. This protein is neither processed to maturity nor sensitive to proteinase K in spheroplasts. In contrast, in the five other mutants, the signal peptide is cleaved and the protein is accessible to proteinase K added to spheroplasts. This indicates that the five mutant proteins are, at least in part, exported through the inner membrane. We propose that the corresponding mutations define two regions of the mature protein (between residues 18 and 42 and between residues 280 and 306), which are important for release of the protein from the inner membrane into the periplasm. We discuss the results in terms of possible conformational changes at this late step of export to the periplasm. PMID- 3049531 TI - Influence of nar (nitrate reductase) genes on nitrate inhibition of formate hydrogen lyase and fumarate reductase gene expression in Escherichia coli K-12. AB - In Escherichia coli, aerobiosis inhibits the synthesis of enzymes for anaerobic respiration (e.g., nitrate reductase and fumarate reductase) and for fermentation (e.g., formate-hydrogen lyase). Anaerobically, nitrate induces nitrate reductase synthesis and inhibits the formation of both fumarate reductase and formate hydrogen lyase. Previous work has shown that narL+ is required for the effects of nitrate on synthesis of both nitrate reductase and fumarate reductase. Another gene, narK (whose function is unknown), has no observable effect on formation of these enzymes. We report here our studies on the role of nar genes in fumarate reductase and formate-hydrogen lyase gene expression. We observed that insertions in narX (also of unknown function) significantly relieved nitrate inhibition of fumarate reductase gene expression. This phenotype was distinct from that of narL insertions, which abolished this nitrate effect under certain growth conditions. In contrast, insertion mutations in narK and narGHJI (the structural genes for the nitrate reductase enzyme complex) significantly relieved nitrate inhibition of formate-hydrogen lyase gene expression. Insertions in narL had a lesser effect, and insertions in narX had no effect. We conclude that nitrate affects formate-hydrogen lyase synthesis by a pathway distinct from that for nitrate reductase and fumarate reductase. PMID- 3049533 TI - Feedback regulation of the spc operon in Escherichia coli: translational coupling and mRNA processing. AB - The spc operon of Escherichia coli encodes 10 ribosomal proteins in the order L14, L24, L5, S14, S8, L6, L18, S5, L30, and L15. This operon is feedback regulated by S8, which binds near the translation start site of L5 and inhibits translation of L5 directly and that of the distal genes indirectly. We constructed plasmids carrying a major portion of the spc operon genes under lac transcriptional control. The plasmids carried a point mutation in the S8 target site which abolished regulation and resulted in overproduction of plasmid-encoded ribosomal proteins upon induction. We showed that alteration of the AUG start codon of L5 to UAG decreased the synthesis rates of plasmid-encoded distal proteins, as well as L5, by approximately 20-fold, with a much smaller (if any) effect on mRNA synthesis rates, indicating coupling of the distal cistrons' translation with the translation of L5. This conclusion was also supported by experiments in which S8 was overproduced in trans. In this case, there was a threefold reduction in the synthesis rates of chromosome-encoded L5 and the distal spc operon proteins, but no decrease in the mRNA synthesis rate. These observations also suggest that transcription from ribosomal protein promoters may be special, perhaps able to overcome transcription termination signals. We also analyzed the state of ribosomal protein mRNA after overproduction of S8 in these experiments and found that repression of ribosomal protein synthesis was accompanied by stimulation of processing (and degradation) of spc operon mRNA. The possible role of mRNA degradation in tightening the regulation is discussed. PMID- 3049535 TI - Aspartate taxis mutants of the Escherichia coli tar chemoreceptor. AB - The Tar protein of Escherichia coli belongs to a family of methyl-accepting inner membrane proteins that mediate chemotactic responses to a variety of compounds. These transmembrane signalers monitor the chemical environment by means of specific ligand-binding sites arrayed on the periplasmic side of the membrane, and in turn control cytoplasmic signals that modulate the flagellar rotational machinery. The periplasmic receptor domain of Tar senses two quite different chemoeffectors, aspartate and maltose. Aspartate is detected through direct binding to Tar molecules, whereas maltose is detected indirectly when complexed with the periplasmic maltose-binding protein. Saturating levels of either aspartate or maltose do not block behavioral responses to the other compound, indicating that the detection sites for these two attractants are not identical. We initiated structure-function studies of these chemoreceptor sites by isolating tar mutants which eliminate aspartate or maltose taxis, while retaining the ability to respond to the other chemoeffector. Mutants with greatly reduced aspartate taxis are described and characterized in this report. When present in single copy in the chromosome, these tar mutations generally eliminated chemotactic responses to aspartate and structurally related compounds, such as glutamate and methionine. Residual responses to these compounds were shifted to higher concentrations, indicating a reduced affinity of the aspartate-binding site in the mutant receptors. Maltose responses in the mutants ranged from 10 to 80% of normal, but had no detectable threshold shifts, indicating that these receptor alterations may have little effect on maltose detection sensitivity. The mutational changes in 17 mutants were determined by DNA sequence analysis. Each mutant exhibited a single amino acid replacement at residue 64, 69, or 73 in the Tar molecule. The wild-type Tar transducer contains arginines at all three of these positions, implying that electrostatic forces may play an important role in aspartate detection. PMID- 3049534 TI - Analysis of the Pseudomonas solanacearum polygalacturonase encoded by pglA and its involvement in phytopathogenicity. AB - A major endopolygalacturonase excreted by Pseudomonas solanacearum was purified to greater than 95% homogeneity and shown to have an isoelectric point of 9.0 and a subunit molecular mass of 52 kilodaltons (kDa). The gene encoding this enzyme (pglA) was isolated from a genomic library of P. solanacearum DNA based on its expression in Escherichia coli and shown to be contained on a 1.8-kilobase DNA fragment. The identity of the pglA gene product and the 52-kDa polygalacturonase was demonstrated by immunoadsorption and isoelectric focusing experiments. The cloned pglA gene was apparently expressed from its own promoter in E. coli and its product was partially secreted into the periplasm. The pglA gene was insertionally inactivated in vitro and used to mutate the chromosomal pglA gene of P. solanacearum by marker exchange mutagenesis. The resulting mutant strain was deficient in production of the 52-kDa polygalacturonase and took twice as long to wilt and kill tomato plants as the wild-type parent in plant bioassay experiments. Complementation in trans with the wild-type cloned pglA gene restored virulence to near wild-type levels. The data indicate that the pglA gene is important, but not absolutely necessary, for pathogenesis. PMID- 3049536 TI - Maltose chemoreceptor of Escherichia coli: interaction of maltose-binding protein and the tar signal transducer. AB - The maltose chemoreceptor in Escherichia coli consists of the periplasmic maltose binding protein (MBP) and the Tar signal transducer, which is localized in the cytoplasmic membrane. We previously isolated strains containing malE mutations that cause specific defects in the chemotactic function of MBP. Four of these mutations have now been characterized by DNA sequence analysis. Two of them replace threonine at residue 53 of MBP with isoleucine (MBP-TI53), one replaces an aspartate at residue 55 with asparagine (MBP-DN55), and the fourth replaces threonine at residue 345 with isoleucine (MBP-TI345). The chemotactic defects of MBP-TI53 and MBP-DN55, but not of MBP-TI345, are suppressed by mutations in the tar gene. Of the tar mutations, the most effective suppressor (isolated independently three times) replaces Arg-73 of Tar with tryptophan. Two other tar mutations that disrupt the aspartate chemoreceptor function of Tar also suppress the maltose taxis defects associated with MBP-TI53 and MBP-DN55. One of these mutations introduces glutamine at residue 73 of Tar, the other replaces arginine at residue 69 of Tar with cysteine. These results suggest that regions of MBP that include residues 53 to 55 and residue 345 are important for the interaction with Tar. In turn, arginines at residues 69 and 73 of Tar must be involved in the recognition of maltose-bound MBP and/or in the production of the attractant signal generated by Tar in response to maltose-bound MBP. PMID- 3049537 TI - Isolation, hyperexpression, and sequencing of the aceA gene encoding isocitrate lyase in Escherichia coli. AB - A structural gene for isocitrate lyase was isolated from a cosmid containing an ace locus of the Escherichia coli chromosome. Cloning and expression under control of the tac promoter in a multicopy plasmid showed that a 1.7-kilobase pair DNA segment was sufficient for complementation of an aceA deletion mutation and overproduction of isocitrate lyase. DNA sequence analysis of the cloned gene and N-terminal protein sequencing of the cloned and wild-type enzymes revealed an entire aceA gene which encodes a 429-amino-acid residue polypeptide whose C terminus is histidine. The deduced amino acid sequence for the 47.2-kilodalton subunit of E. coli isocitrate lyase could be aligned with that for the 64.8 kilodalton subunit of the castor bean enzyme with 39% identity except for limited N- and C-terminal regions and a 103-residue stretch that was unique for the plant enzyme and started approximately in the middle of that peptide. PMID- 3049538 TI - Rapid and precise mapping of the Escherichia coli release factor genes by two physical approaches. AB - The termination of protein synthesis in Escherichia coli requires two codon specific factors termed RF1 and RF2. RF1 mediates UAA- and UAG-directed termination, while RF2 mediates UAA- and UGA-directed termination. The genes encoding these factors have been isolated and sequenced, and RF2 was found to be encoded in two separate reading frames. The map position of RF1 has been reported as 27 min on the E. coli chromosome, while the RF2 map position has not yet been identified. In this study, two new and independent methods for gene mapping, using pulsed field gel electrophoresis and an ordered bacteriophage library spanning the entire chromosome, were used to localize the map position of the RF2 gene. In addition, the location of the RF1 gene was more precisely defined. The RF2 gene is located at 62.3 min on the chromosome, while the RF1 gene is located at 26.7 min. This approach to mapping cloned genes promises to be a rapid and simple means for determining the gene order of the genome. PMID- 3049539 TI - Nucleotide sequence of the xth gene of Escherichia coli K-12. AB - The xth gene of Escherichia coli K-12, which encodes exonuclease III, has been sequenced. Exonuclease III from a cloned copy of the E. coli K-12 gene has been purified and characterized. The molecular weight (30,921), the amino-terminal amino acid sequence, and the amino acid composition of the polypeptide predicted from the nucleotide sequence are in excellent agreement with those properties determined for the purified enzyme. The xth promoter was mapped by primer extension of in vivo transcripts. Inspection of the nucleotide sequence reveals that a region of dyad symmetry which could form a hairpin stem-loop structure in RNA characteristic of a rho-dependent terminator lies immediately downstream from the xth gene. PMID- 3049540 TI - Microcalorimetric monitoring of growth of Saccharomyces cerevisiae: osmotolerance in relation to physiological state. AB - The importance of the physiological state of a culture of Saccharomyces cerevisiae for tolerance to sudden osmotic dehydration was studied, and it was investigated whether specific osmotolerance factors were demonstrable. The microcalorimeter was used to monitor growth, and different physiological states of the culture were selected and their osmotolerance was tested. In addition to cells in the stationary phase, cells from the transition phase between respirofermentative and respiratory catabolism were osmotolerant. S. cerevisiae exhibited ever-changing metabolism during batch growth on either glucose or ethanol as the carbon source. Instantaneous heat production per biomass formation (dQ/dX) and specific activity of sn-glycerol 3-phosphate dehydrogenase (GPDH) (EC 1.1.1.8) were shown to differ for different physiological states. Neither high respiratory activity nor low total cellular activity, nor factors involved in osmoregulation, i.e., intracellular glycerol or activity of GPDH, correlated with the osmotolerant phenotype. PMID- 3049542 TI - Determinations of the DNA sequence of the mreB gene and of the gene products of the mre region that function in formation of the rod shape of Escherichia coli cells. AB - The 6.5-kilobase mre region at 71 min in the Escherichia coli chromosome map, where genes involved in formation of a rod-shaped cell form a gene cluster, was analyzed by in vivo protein synthesis in a maxicell system and by base sequencing of DNA. An open reading frame that may code for a protein with an Mr of about 37,000 on sodium dodecyl sulfate-polyacrylamide gels was found and was correlated with the mreB gene. N-terminal amino acid sequencing of the hybrid mreB-lacZ protein confirmed the production by mreB of a protein of 347 amino acid residues with a molecular weight of 36,958. The amino acid sequence of this protein deduced from the DNA sequence showed close similarity with that of a protein of the ftsA gene which is involved in cell division of E. coli. Three other contiguous genes that formed three proteins with Mrs of about 40,000, 22,000, and 51,000, respectively, were detected downstream of the mreB gene by in vivo protein synthesis. The mreB protein and some of these three proteins may function together in determination of cell shape. PMID- 3049541 TI - Overproduction of MalK protein prevents expression of the Escherichia coli mal regulon. AB - The mal regulon of Escherichia coli comprises a large family of genes whose function is the metabolism of linear maltooligosaccharides. Five gene products are required for the active accumulation of maltodextrins as large as maltoheptaose. Two cytoplasmic gene products are necessary and sufficient for the intracellular catabolism of these sugars. Two newly discovered enzymes have the capacity to metabolize these sugars but are not essential for their catabolism in wild-type cells. A single regulatory protein, MalT, positively regulates the expression of all of these genes in response to intracellular inducers, one of which has been identified as maltotriose. In the course of studying the mechanism of the transport system, we have placed the structural gene for one of the transport proteins, MalK, under the control of the Ptrc promoter to produce large amounts of this protein. We found that although high-level expression of MalK was not detrimental to E. coli, the increased amount of MalK decreased the basal level expression of the mal regulon and prevented induction of the mal system even in the presence of external maltooligosaccharides. Constitutive mutants in which MalT does not depend on the presence of the internal inducer(s) were unaffected by the increased levels of the MalK protein. These results are consistent with the idea that MalK protein somehow interferes with the activity of the MalT protein. Different models for the regulatory function of MalK are discussed. PMID- 3049544 TI - Role of cloned carotenoid genes expressed in Escherichia coli in protecting against inactivation by near-UV light and specific phototoxic molecules. AB - Genes controlling carotenoid synthesis were cloned from Erwinia herbicola and expressed in an Escherichia coli strain. Carotenoids protect against high fluences of near-UV (NUV; 320 to 400 nm) but not against far-UV (200-300 nm). Protection of E. coli cells was not observed following treatment with either psoralen or 8-methoxypsoralen plus NUV. However, significant protection of cells producing carotenoids was observed with three photosensitizing molecules activated by NUV (alpha-terthienyl, harmine, and phenylheptatriyne) which are thought to have the membrane as an important lethal target. Protection of carotenoid-producing cells against inactivation was not observed with acridine orange plus visible light but was seen with toluidine blue O plus visible light. PMID- 3049543 TI - Molecular cloning, heterologous expression, and primary structure of the structural gene for the copper enzyme nitrous oxide reductase from denitrifying Pseudomonas stutzeri. AB - The nos genes of Pseudomonas stutzeri are required for the anaerobic respiration of nitrous oxide, which is part of the overall denitrification process. A nos coding region of ca. 8 kilobases was cloned by plasmid integration and excision. It comprised nosZ, the structural gene for the copper-containing enzyme nitrous oxide reductase, genes for copper chromophore biosynthesis, and a supposed regulatory region. The location of the nosZ gene and its transcriptional direction were identified by using a series of constructs to transform Escherichia coli and express nitrous oxide reductase in the heterologous background. Plasmid pAV5021 led to a nearly 12-fold overexpression of the NosZ protein compared with that in the P. stutzeri wild type. The complete sequence of the nosZ gene, comprising 1,914 nucleotides, together with 282 nucleotides of 5' flanking sequences and 238 nucleotides of 3'-flanking sequences was determined. An open reading frame coded for a protein of 638 residues (Mr, 70,822) including a presumed signal sequence of 35 residues for protein export. The presequence is in conformity with the periplasmic location of the enzyme. Another open reading frame of 2,097 nucleotides, in the opposite transcriptional direction to that of nosZ, was excluded by several criteria from representing the coding region for nitrous oxide reductase. Codon usage for nosZ of P. stutzeri showed a high G + C content in the degenerate codon position (83.9% versus an average of 60.2%) and relaxed codon usage for the Glu codon, characteristic features of Pseudomonas genes from other species. E. coli nitrous oxide reductase was purified to homogeneity. It had the Mr of the P. stutzeri enzyme but lacked the copper chromophore. PMID- 3049546 TI - Effects of release factor context at UAA codons in Escherichia coli. AB - In Escherichia coli, nonsense suppression at UAA codons is governed by the competition between a suppressor tRNA and the translational release factors RF1 and RF2. We have employed plasmids carrying the genes for RF1 and RF2 to measure release factor preference at UAA codons at 13 different sites in the lacI gene. We show here that the activity of RF1 and RF2 varies according to messenger context. RF1 is favored at UAA codons which are efficiently suppressed. RF2 is preferred at poorly suppressed sites. PMID- 3049545 TI - Transcriptional analysis of the major surface array gene of Caulobacter crescentus. AB - The major component of the paracrystalline surface array of Caulobacter crescentus CB15 and one of the most abundant cellular proteins is a protein designated 130K. We have determined the DNA sequence of the 5' portion of the 130K gene, including the N-terminal one-third of the protein coding region, and analyzed the transcription of the gene. The site of transcription initiation was determined by S1 mapping of Caulobacter RNA. Although the DNA sequence upstream from the transcription start site showed significant homology to the consensus promoter sequences of Escherichia coli, S1 analysis of RNA from E. coli carrying the 130K gene on a plasmid indicated that the 130K promoter was not transcribed by E. coli RNA polymerase in vivo. Quantitative S1 analysis of RNA isolated from synchronously growing Caulobacter cells suggested that this promoter was not under developmental regulation; the amount of 130K transcript varied no more than 1.5-fold during the cell cycle. The length of the 130K mRNA was determined to be 3.3 kilobases by Northern (RNA blot) analysis, indicating that the 130K mRNA is not part of a polycistron. The amino acid sequence predicted from the DNA sequence agreed well with the N-terminal amino acid sequence determined by sequencing of the 130K protein. The 130K protein appears to be synthesized without an N-terminal leader sequence, but the N-terminal 20 amino acids are relatively hydrophobic and may function like a signal sequence during transmembrane translocation. PMID- 3049547 TI - Differences in penicillin-binding proteins of Streptococcus pyogenes and two derived, stabilized L forms. AB - The penicillin-binding proteins (PBPs) of Streptococcus pyogenes and two of its derived, stabilized (i.e., nonreverting) L forms, an osmotically fragile L form and a physiologic isotonic L form, were compared. The numbers of PBPs in the membranes of these organisms were 6, 4, and 2 for the coccus and the osmotically fragile and physiologic isotonic L forms, respectively. Likewise, the relative amounts of total PBPs were 1.00: 1.48:0.32 for this coccus and the osmotically fragile and physiologic isotonic L forms, respectively. The two largest PBPs (PBPs 1 and 2) of the coccus were absent in both L forms, while the smallest PBPs (PBPs 5 and 6) were found in all three membranes. Deacylation (half-life) of three of the four PBPs in the osmotically fragile L form membrane required a significantly longer time than did deacylation of these presumed identical enzymes in the parental coccal membrane. Conversely, there was no such difference between the only two PBPs of the physiologic isotonic L form and the same coccal membrane proteins. Intact cells of all three organisms secreted PBPs and what appeared to be penicilloic acid and a minimal amount of free penicillin. A greater amount of these PBPs was secreted by both L forms than by the coccus. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis patterns and ratios of secreted PBPs were identical to those from labeled membrane preparations. These differences are correlated with some of our previous findings and are discussed in terms of inhibition of cell wall synthesis and resulting membrane changes in these two derived, stabilized coccal L forms. PMID- 3049548 TI - Translational coupling between the ilvD and ilvA genes of Escherichia coli. AB - The hypothesis that translation of the ilvD and ilvA genes of Escherichia coli may be linked has been examined in strains in which lacZ-ilvD protein fusions are translated in all three reading frames with respect to ilvD. In these strains, the nucleotide sequence was altered to obtain premature termination of ilvD translation, and in one strain translation termination of ilvD DNA occurred two bases downstream of the ilvA initiation codon. In the wild-type strain, the ilvD translation termination site was located two bases upstream of the ilvA start codon. In each of the mutant strains, expression of ilvA, as determined by the level of threonine deaminase activity, was strikingly lower than in the wild-type strain. The data suggest that expression of ilvD and ilvA is translationally coupled. By inserting a promoterless cat gene downstream of ilvA, it was shown that the differences in enzyme activity were not the result of differences in the amount of ilvA mRNA produced. PMID- 3049549 TI - Activation of protease-constitutive recA proteins of Escherichia coli by all of the common nucleoside triphosphates. AB - To understand why the RecA proteins of the protease-constitutive recA1202 and recA1211 mutants show very high protease activities in vivo without the usual need for DNA damage (E. S. Tessman and P. Peterson, J. Bacteriol. 163:677-687, 1985), we examined the activation of the mutant proteins by nucleoside triphosphates (NTPs) in vitro. In vivo, the mutant protease activities are resistant to inhibition by cytidine plus guanosine (C + G) in the growth medium, in contrast to the activities of weaker mutants, such as recA441, which are sensitive to C + G inhibition. We found that RecA1202 and RecA1211 proteins, in contrast to RecA+, can use natural NTPs other than ATP and dATP as cofactors in the cleavage of LexA repressor. The effectiveness of NTPs in promoting LexA cleavage by RecA1202 and RecA1211 proteins decreased in roughly the following order: dATP greater than ATP greater than UTP greater than ATP-gamma S greater than dCTP greater than CTP greater than dGTP greater than GTP greater than TTP. These mutant proteins showed higher affinities for ATP and single-stranded DNA and higher repressor cleavage activities than RecA+ protein. With the various effectors (single-stranded DNA or NTPs), the RecA1202 protein always showed more activity than RecA1211 in the cleavage of LexA repressor in vitro, which is consistent with the greater activity of the recA1202 mutant in vivo. The results explain, in part, why some recA mutants have unusually high constitutive RecA protease activity and why that activity is more or less resistant to C + G inhibition. PMID- 3049550 TI - An amino acid substitution in penicillin-binding protein 3 creates pointed polar caps in Escherichia coli. AB - The pbpB gene product penicillin-binding protein 3 (PBP3) of Escherichia coli is one of the major targets of beta-lactam antibiotics. At the permissive temperature, the temperature-sensitive pbpBr1 mutant, which was obtained after selection for increased resistance to cephalexin, shows a dramatic change in shape which has never been observed before; the polar caps are pointed. We show that the substitution of amino acid Asn-361 by Ser, previously shown to be responsible for increased cephalexin resistance and for temperature sensitivity, causes the pointed polar caps. However, comparison of the morphological and physiological characteristics of the pbpBr1 mutant with those of other pbpB mutants suggests that the formation of pointed polar caps is not correlated with temperature sensitivity or cephalexin resistance. Partial inactivation of PBP3 by subinhibitory concentrations of cephalexin, furazlocillin, and piperacillin resulted in the formation of slightly pointed polar caps, suggesting that the shape of the polar caps is correlated with PBP3 activity. The large change in the shape of the polar caps was accompanied by a small change in the kinetics of peptidoglycan synthesis and in the local rate of surface synthesis activity along the cell envelope. PMID- 3049551 TI - High-affinity glucose transport in Saccharomyces cerevisiae is under general glucose repression control. AB - Saccharomyces cerevisiae mutants defective in growth on low glucose concentration (lgn mutants) were isolated and screened for abnormal glucose transport. Nine complementation groups were identified, falling into two broad groups: those unable to significantly derepress high-affinity (low-Km) glucose uptake (lgn1, lgn4, lgn5, lgn7, and lgn8), and those with elevated repressed levels of high affinity uptake that either derepress to normal or near normal levels of high affinity uptake with loss of low-affinity transport (lgn2 and lgn3) or derepress only slightly, appearing to have an intermediate yet constitutive level of high affinity transport (lgn6 and lgn9). Further analysis of the lgn mutations revealed pleiotropic phenotypes most consistent with the true defect being in regulation or expression of glucose repression and derepression. The kinetics of glucose uptake in strains carrying known mutations preventing derepression of glucose-repressible functions (snf1, snf2, snf4, and snf6) demonstrated that three of these mutations (snf1, snf4, and snf6) were similarly defective in derepression of high-affinity glucose uptake. The snf2 and snf5 mutations had no apparent effect on glucose uptake. Two mutations resulting in constitutive expression of glucose-repressible functions, cid1 and reg1, resulted in constitutive expression of high-affinity glucose uptake. These data support the conclusion that high-affinity glucose uptake in Saccharomyces cerevisiae is under general glucose repression control. The implications of other properties of these mutants are discussed. PMID- 3049552 TI - Characterization and expression of two avirulence genes cloned from Pseudomonas syringae pv. glycinea. AB - Two avirulence genes, avrB and avrC, from race 0 of Pseudomonas syringae pv. glycinea, were sequenced and found to encode single protein products of 36 and 39 kilodaltons, respectively. The proteins had neither recognizable signal peptide sequences nor significant stretches of hydrophobic amino acids that might indicate membrane association. Both avrB and avrC had relatively low position 3 and overall G+C contents, which suggests that they may have been recently introduced into P. syringae pv. glycinea. The deduced amino acid sequences of the proteins encoded by avrB and avrC shared 42% identical amino acids. However, when introduced into race 4 of P. syringae pv. glycinea, each gene directed a unique pattern of hypersensitive reactions on several differential soybean cultivars. The avrC protein was overproduced in Escherichia coli cells and deposited as insoluble inclusion bodies in the cell cytoplasm. The avrC protein could be solubilized with urea-octyl glucoside treatment, but neither the solubilized protein nor the intact inclusion bodies elicited a hypersensitive reaction in soybean leaves. PMID- 3049554 TI - 31P nuclear magnetic resonance studies of effects of some chlorophenols on Escherichia coli and a pentachlorophenol-degrading bacterium. AB - A Flavobacterium sp. that mineralizes pentachlorophenol degrades some, but not all, of the other chlorinated phenols. Whole-cell 31P nuclear magnetic resonance was used to compare and observe transmembrane pH gradients and nucleotide pools in the Flavobacterium sp. and Escherichia coli after pentachlorophenol and 3,4,5 trichlorophenol were added to the cell suspensions. The data suggest that those chlorinated phenols which are not degraded by the Flavobacterium sp. may be resistant to degradation because they act as proton dissipators. PMID- 3049556 TI - Fine structures of the capsules of Klebsiella pneumoniae and Escherichia coli K1. AB - The fine structures of the capsules of Klebsiella pneumoniae and Escherichia coli were determined by the rapid-freezing technique. The capsular layer was seen as a densely packed accumulation of fine fibers. The thickness of the capsule was approximately 160 nm in K. pneumoniae and less than 10 nm in E. coli K1. Two layers were observed in the Klebsiella capsule in which the arrangements of the fibers were different. The inner layer of the capsule was formed by a palisade of thick and dense bundles of the fibers standing at right angles on the surface of the outer membrane. In the outer layer these thick bundles of fibers loosened into fine fibers which spread over the bacterial surface, forming a fine network structure. PMID- 3049555 TI - Direction of conjugative transfer of IncI1 plasmid ColIb-P9. AB - The origin-of-transfer region of ColIb-P9 was inserted into a lambda prophage to give a bacterial chromosome mobilizable by the parental conjugative plasmid. The polarity of mobilization of chromosomal genes indicated that ColIb-P9 transfer is unidirectional, such that the transfer genes adjacent to oriT enter the recipient cell last. PMID- 3049553 TI - DNA sequence of the gene (tyrP) encoding the tyrosine-specific transport system of Escherichia coli. AB - The nucleotide sequence of 1,947 bases of DNA containing the tyrP structural gene was determined, and an open reading frame of 1,260 nucleotides was identified. The putative structural gene encodes an extremely hydrophobic protein which comprises 404 amino acids, 70% of which are nonpolar, and which has a molecular weight of 43,261. PMID- 3049558 TI - Availability of porphobilinogen controls appearance of porphobilinogen deaminase activity in Escherichia coli K-12. AB - A hemin-permeable hemB mutant had no 5-aminolevulinate dehydratase (ALA D) and extremely low porphobilinogen deaminase (PBG D) activity. When the structural gene for hemB was introduced into this strain on a single-copy plasmid, both activities were observed. When the mutant was grown on PBG, normal PBG D activity was observed. Moreover, a hemA mutant had little or no PBG D activity unless it was grown on ALA or PBG. Neither hemin nor PBG affected the level of PBG D protein produced from in vitro transcription and translation of a plasmid harboring the hemC gene as an insert. We conclude that, in Escherichia coli, PBG availability controls the activity of PBG D at some posttranscriptional level. PMID- 3049557 TI - Very short patch mismatch repair activity associated with gene dcm is not conferred by a plasmid coding for EcoRII methylase. AB - The only cytosine methylase in Escherichia coli K-12 methylates the second cytosine in the sequence CC (A/T)GG and is encoded by gene dcm. Methylation and very short patch mismatch repair activities lacking in a dcm mutant of E. coli were restored by a plasmid containing the cloned dcm gene. In contrast, plasmids with the gene for EcoRII methylase, which is a homolog of dcm, restored only cytosine methylase activity and not mismatch repair. PMID- 3049559 TI - The dexamethasone suppression test in children and adolescents with major depressive disorder: a review. AB - The authors review the available studies on the use of the dexamethasone suppression test (DST) in children and adolescents with psychiatric disorders. For the purposes of analysis, the authors divided the studies according to the two age groups. Pooled sensitivity, specificity, and positive predictive power were calculated for each age group per diagnostic category. DST nonsuppression among subjects with major depressive disorder (MDD) in the children's group was 69.6%, with a specificity of 69.7%, whereas for adolescent subjects, DST sensitivity was 41.3%, and specificity was 79.3%. The positive predictive power of an abnormal DST result in helping diagnose MDD in the children's group was 69.7%, whereas for the adolescent group, the positive predictive power was 66.6%. The authors conclude that the evidence cannot support the routine clinical use of the DST in children and in adolescents, but there may be a research role for the DST in these patient populations. PMID- 3049560 TI - Clinical outcome and adverse effect profile associated with concurrent administration of alprazolam and imipramine. AB - Clinical outcome and adverse effects associated with concurrent alprazolam and imipramine administration were studied in 29 patients with major depressive disorder who completed a 6-week trial in which they served as their own controls. Alprazolam was added on Day 8 in gradually escalating, then gradually tapering dosages while imipramine dosages remained unchanged. Significant decreases were observed in scores on the Hamilton Rating Scales for Depression and Anxiety at all later evaluation days with Day 8 as baseline. The mean total Symptom and Side Effects score decreased significantly from Day 8 to Day 22 when alprazolam doses were 1 mg q.i.d. For most side effects, total number of reports remained constant or decreased from Day 1 to later evaluation days. Standing diastolic blood pressures were significantly lower on Day 22 than on Day 1. No significant relationship was found between any rating scale score and plasma concentration data. PMID- 3049562 TI - Anxiety and alcoholism. AB - The prevalence and the co-occurrence of anxiety disorders and alcoholism in individuals, families, and the general population have been documented in both epidemiologic and clinical investigations. In patients who fulfill the criteria for one of those conditions, clinicians should consider the possibility of the presence of the other disorder--currently, at a previous time, or at some point in the future. A lifetime history of alcohol abuse and anxiety disorders obtained from both the patient and the family members may assist in determining the appropriate treatment. PMID- 3049561 TI - Long-term outpatient treatment of senile dementia with oral physostigmine. AB - The authors describe five patients with senile dementia of the Alzheimer type (SDAT) who were treated with oral physostigmine from 1 to 3 3/4 years. An abbreviated form of the Buschke selective reminding test was used to assess the patients' short-term verbal memory at periodic clinic visits. Although one of the patients deteriorated progressively from the start of the physostigmine therapy and was taken off the drug after 1 year, there was no evidence of deterioration in memory performance on the average in the other four patients. Equally important, this open trial provides evidence of the safety of long-term treatment with physostigmine on an outpatient basis. The authors view their results as supporting the potential value of cholinesterase inhibition as a means of forestalling memory decline in SDAT patients. PMID- 3049563 TI - Suicide and other consequences of childhood and adolescent anxiety disorders. AB - The increasing risk of suicide among children and adolescents makes imperative the development of a set of appropriate and clinically significant predictors of suicide risk for that group. Of particular concern is the need to understand the role of anxiety psychopathology in suicidal children and the occurrence of suicidal behaviors in children with anxiety disorders. The author reviews the recent investigations assessing childhood and adolescent suicide risk and emphasizes the need to update our current methods in assessing both suicidal behavior and anxiety. Furthermore, he points out the need for prospective studies of anxiety disorders in children and adolescents to identify those children at risk for suicide and other psychopathology as they mature. PMID- 3049564 TI - Characteristic distribution of cathepsin E which immunologically cross-reacts with the 86-kDa acid proteinase from rat gastric mucosa. AB - The antiserum raised against the high-molecular-weight acid proteinase from rat gastric mucosa, termed 86-kDa acid proteinase, has been shown to recognize rat cathepsin E, but not cathepsin D (Muto, N. et al. (1987) J. Biochem. 101, 1069 1075). Using this specific antiserum, characteristic distribution of cathepsin E in rats was demonstrated. The enzyme was detected in a limited number of tissues, such as stomach, thymus, spleen, bladder, and erythrocyte membranes. Among them, the highest activity was observed in the stomach. In contrast, cathepsin D immunoreactive with the antiserum specific to rat gastric cathepsin D was demonstrated in all the tissues examined. Cathepsin E-type enzymes partially purified from these five tissues were precipitated in the same manner by the specific antiserum, and they had the same molecular weight, electrophoretic mobility, and resistance against denaturation by 4 M urea. These results indicate that they could be exactly classified as cathepsin E. This type of enzyme was also detectable in mice and guinea pigs, but they showed relatively weak immunoreactivities with the antiserum. Thus, it is concluded that the distribution of cathepsin E is intrinsically different from ordinary cathepsin D, suggesting that it has a different physiological role from cathepsin D. PMID- 3049565 TI - Characterization and partial purification of a non-interferon macrophage activating factor produced by human leukemic T cell line. AB - Culture supernatants from several human leukemic T cell lines were found to contain a macrophage activating factor which enhanced hydrogen peroxide release from human peripheral blood monocyte-derived macrophages. The macrophage activating factor from a T cell line, CCRF-CEM, was characterized biochemically and compared with interferon-gamma, which is also an immunological product of T cells and has a potent macrophage activating activity. In contrast to interferon gamma, the macrophage activating factor in the culture supernatants bound to an anion exchanger and did not adsorb onto concanavalin A gel. Culture supernatants and active fractions from chromatographies were essentially devoid of anti-viral activity. Anti-human interferon-gamma monoclonal antibody also failed to neutralize the macrophage activating factor from CCRF-CEM. MAF was eluted in the fractions with molecular weight of 40,000 to 60,000 on gel filtration in the presence of a detergent and a salt. MAF was partially purified to about 1,300 fold by the methods described above: chromatography with anion exchangers and gel filtration. It was concluded that MAF from CCRF-CEM was biochemically and immunologically different from interferon-gamma. PMID- 3049566 TI - Inhibitory effect of sarcotoxin IIA, an antibacterial protein of Sarcophaga peregrina, on growth of Escherichia coli. AB - The effect of sarcotoxin IIA, an antibacterial protein of Sarcophaga peregrina (flesh fly), on Escherichia coli was investigated. Sarcotoxin IIA was found to have a bacterial effect on growing bacteria, but little on non-growing bacteria. At a concentration of 25 micrograms/ml, it induced significant morphological change of growing E. coli cells. In its presence, growing cells became greatly elongated, and spheroplast-like bulges and projections appeared on their surface. A rough mutant strain of E. coli with a defect in the structure of lipopolysaccharide was more sensitive than the parent strain to sarcotoxin IIA. These results suggest that the main effect of sarcotoxin IIA is to inhibit cell wall synthesis, including septum formation. PMID- 3049568 TI - Effects of polypeptide growth factors on mandibular condylar cartilage of the rat in vitro. AB - In a comparative study, the influence of several polypeptide growth factors on proliferation and matrix synthesis in secondary mandibular condylar and primary costal cartilage of the rat were determined using a serum-free culture system. Somatomedin-C, multiplication-stimulating activity (MSA), epidermal growth factor (EGF) and insulin had a significant dose-dependent stimulating effect on proliferation in mandibular condylar cartilage. In costal cartilage, the same factors as well as parathyroid hormone (fragment 1-34), platelet-derived growth factor (PDGF) and high doses of human growth hormone (1 microgram/ml) significantly stimulated proliferation. Matrix synthesis in both cartilages could only be stimulated by high doses of insulin (100 micrograms/ml) and in costal cartilage also by parathyroid hormone. In this culture system fibroblast growth factor reduced proliferation and matrix production, while cartilage-derived factor had no marked effect on the growth processes in both cartilage types. Prominent differences between condylar and costal cartilage were demonstrated by the effects of parathyroid hormone and fetal calf serum. Although the effects on matrix synthesis were very moderate in the tissue culture system used, this study demonstrates that most factors conducive to growth in primary cartilage also stimulate growth in condylar cartilage only exposure to growth substances that interfere with the differentiation of prechondroblasts into chondroblasts, a process that is specific for appositionally growing secondary cartilage, may result in different responses between primary and secondary cartilage. PMID- 3049567 TI - The protein responsible for center A/B in spinach photosystem I: isolation with iron-sulfur cluster(s) and complete sequence analysis. AB - The 9 kDa polypeptide from spinach photosystem I (PS I) complex was isolated with iron-sulfur cluster(s) by an n-butanol extraction procedure under anaerobic conditions. The polypeptide was soluble in a saline solution and contained non heme irons and inorganic sulfides. The absorption spectrum of this iron-sulfur protein was very similar to those of bacterial-type ferredoxins. The amino acid sequence of the polypeptide was determined by using a combination of gas-phase sequencer and conventional procedures. It was composed of 80 amino acid residues giving a molecular weight of 8,894, excluding iron and sulfur atoms. The sequence showed the typical distribution of cysteine residues found in bacterial-type ferredoxins and was highly homologous (91% homology) to that deduced from the chloroplast gene, frxA, of liverwort, Marchantia polymorpha. The 9 kDa polypeptide is considered to be the iron-sulfur protein responsible for the electron transfer reaction in PS I from center X to [2Fe-2S] ferredoxin, namely a polypeptide with center(s) A and/or B in PS I complex. It is noteworthy that the 9 kDa polypeptide was rather hydrophilic and a little basic in terms of the primary structure. A three-dimensional structure was simulated on the basis of the tertiary structure of Peptococcus aerogenes [8Fe-8S] ferredoxin, and the portions in the molecule probably involved in contacting membranes or other polypeptides were indicated. The phylogenetic implications of the structure of the present polypeptide as compared with those of several bacterial-type ferredoxins are discussed. PMID- 3049569 TI - Fibronectin-degrading activity in human crevicular fluid, gingival explants culture medium and bacterial plaques. AB - 125I-fibronectin was incubated with extracts having presumably a proteolytic activity. Plaque from children without gingivitis, plaque from adults with chronic periodontitis, human gingival fluid and pooled culture media of human gingival explants were studied. Proteolysis was usually faster with plaque from patients with adult periodontitis than with plaque from children without gingivitis and the inhibition tests showed that several enzymes were implicated in the process. For the culture medium of gingival explants, the electrophoretic profile of the digestion products of fibronectin was different and showed a decreased activity. Nevertheless the gingival fluid gave a very similar degradation to bacterial plaque. The sulcular content was able to assume enzymatic activity capable of destroying fibronectin. These sulcular activities could be important for bacterial colonisation of sulcular surfaces and perhaps also for fibronectin destruction of periodontal tissues. PMID- 3049570 TI - Characterization and conservation of the inner E2 core domain structure of branched-chain alpha-keto acid dehydrogenase complex from bovine liver. Construction of a cDNA encoding the entire transacylase (E2b) precursor. AB - A cDNA clone encoding the entire transacylase (E2b) precursor of the bovine branched-chain alpha-keto acid dehydrogenase complex has been constructed from two overlapping incomplete cDNA clones which were isolated from a lambda ZAP library prepared from bovine liver poly(A)+ RNA. Nucleotide sequencing indicates that this bovine E2b cDNA insert (bE2-11) is 2701 base pairs in length with an open reading frame of 1446 base pairs. The bE2-11 cDNA insert encodes a leader peptide of 61 residues and a mature E2b polypeptide of 421 amino acid residues with a calculated monomeric molecular mass of 46,518 daltons. The molecular mass of the native E2b component isolated from bovine liver is 1,110,000 daltons as determined by sedimentation equilibrium. This value establishes the 24-subunit octahedral model for the quaternary structure of bovine E2b. The amino-terminal sequences of two tryptic fragments (A and B) of the E2b protein have been determined. Fragment A comprises residues 175 to 421 of the E2b protein and is the inner E2 core domain which contains the transacylase active site. Fragment B, produced by further tryptic cleavage of fragment, comprises residues 205 to 421, but does not have transacylase activity. Both fragments A and B confer the highly assembled 24-mer structure. The primary structure of the inner E2 core domain of bovine E2b (fragment A) is very similar to those of three other E2 proteins (human E2p, Escherichia coli E2p, and E. coli E2k). These similarities suggest that these E2 proteins are structurally and evolutionarily related. PMID- 3049571 TI - Insulin-binding peptide. Design and characterization. AB - The design and characterization of a six-amino acid-containing peptide that binds insulin is described. The amino acid sequence of the insulin-binding peptide (IBP) was determined from the strand of DNA complementary to the strand of DNA coding for the insulin molecule in the domain of the insulin monomer believed to interact with the insulin receptor. The IBP (Cys-Val-Glu-Glu-Ala-Ser) binds specifically to insulin in a saturable manner with a Kd of 3 nM. This binding process is time dependent and slightly temperature dependent, and the peptide appears to interact with insulin near the carboxyl terminus of the B-chain of insulin. Incubation of insulin with the peptide decreases insulin binding to the insulin receptor by 50%, with no effect on the affinity of insulin for the receptor and no effect on cellular insulin-stimulated deoxyglucose uptake. A polyclonal antibody produced against the IBP will inhibit specific insulin binding to intact cells by approximately 50%, with no effects on insulin stimulated glucose uptake. From this data, we suggest that there are at least two domains of the insulin molecule through which it interacts with its receptor, the "binding region" of insulin, which is the domain blocked by the IBP, and the "message region" of insulin, through which insulin not only binds to the receptor, but also generates the cellular signal. PMID- 3049572 TI - The complete amino acid sequence of porcine gastrotropin, an ileal protein which stimulates gastric acid and pepsinogen secretion. AB - The stomach is stimulated by an enterooxyntin factor in a delayed response to feeding, resulting in an increase in both gastric acid and pepsinogen secretion. We have previously reported on the identity of such a factor from the porcine ileum (Wider, M. D., Vinik, A. I., and Heldsinger, A. (1984) Endocrinology 115, 1484-1491). This protein, termed gastrotropin, is localized to the distal region of the ileum where it constitutes less than 0.1% of the cytosolic protein. We have completed the primary structure of porcine gastrotropin by Edman degradation and mass spectrometry. Gastrotropin (Mr = 14,054) contains 127 amino acid residues and has a blocked (acetylated) alanine at its NH2 terminus. The sequence of porcine gastrotropin is similar to rat liver fatty acid-binding protein (FABP), with 44 of 127 residues being identical (35%). Homology with other members of the FABP family is significantly less apparent, with the order of similarity being liver FABP greater than heart FABP greater than retinol-binding protein greater than intestine FABP. The sequences of the NH2-terminal regions of these proteins account for virtually all of the homology; there are 9 conserved residues common to all five proteins. Gastrotropin represents the first member of the FABP family which has an extracellular function. PMID- 3049573 TI - Ribosome release modulates basal level expression of the trp operon of Escherichia coli. AB - The leader peptide stop codon (UGA) of the Escherichia coli trp operon was replaced by UAA and UAG. The transcriptional behavior of the mutated leader regions in vitro and the extent of transcription termination observed with each in vivo were virtually identical to that of the wild type leader region. Introduction of a release factor 1 (UAA- and UAG-specific) mutation into strains with the different stop codons caused increased termination in strains with UAA and UAG, but not with UGA (in cells grown in the presence of tryptophan). This finding provides evidence for the view that ribosome release from the leader peptide stop codon is an important event in setting the basal level of transcription readthrough at the trp attenuator. PMID- 3049574 TI - Chloroplast rpoA, rpoB, and rpoC genes specify at least three components of a chloroplast DNA-dependent RNA polymerase active in tRNA and mRNA transcription. AB - The purpose of this study was to determine the relationship between putative chloroplast RNA polymerase subunit genes and known chloroplast transcriptional activities. We have prepared fusion polypeptide genes from fragments of chloroplast DNA homologous to bacterial RNA polymerase subunit genes and expression vectors carrying portions of the anthranilate synthetase gene (trpE). Fusion proteins for chloroplast homologs of the RNA polymerase alpha (rpoA), beta (rpoB), and beta' (rpoC) subunits were obtained from these genes. The fusion polypeptides synthesized by Escherichia coli in vivo were purified and used as antigens for production of rabbit polyclonal anti-RNA polymerase subunit-specific antibodies. The purified antibodies were able to immobilize chloroplast DNA dependent RNA polymerases from spinach, pea, and Euglena gracilis. In addition, the soluble chloroplast RNA polymerase activity in tRNA and mRNA synthesis was strongly inhibited by these antibodies under conditions which had little effect on transcription by the chloroplast transcriptionally active chromosome that preferentially transcribed rRNA genes (Greenberg, B. M., Narita, J. O., DeLuca Flaherty, C., Gruissem, W., Rushlow, K. A., and Hallick, R. B. (1984) J. Biol. Chem. 259: 14880-14887). From these data we conclude that the chloroplast genes homologous to bacterial RNA polymerase subunit genes are expressed in vivo and that the protein products specify at least three of the components of the chloroplast RNA polymerase(s) involved in tRNA and mRNA transcription. PMID- 3049575 TI - Cloning and expression of the Photobacterium phosphoreum luminescence system demonstrates a unique lux gene organization. AB - The organization of the lux structural genes (A-E) in Photobacterium phosphoreum has been determined and a new gene designated as luxF discovered. The P. phosphoreum luminescence system was cloned into Escherichia coli using a pBR322 vector and identified by cross-hybridization with Vibrio fischeri lux DNA. The lux genes were located by specific expression of P. phosphoreum DNA fragments in the T7-phage polymerase/promoter system in E. coli and identification of the labeled polypeptide products. The luxA and luxB gene products (luciferase subunits) were shown to catalyze light emission in the presence of FMNH2, O2, and aldehyde. The luxC, luxD, and luxE gene products (fatty acid reductase subunits) responsible for aldehyde biosynthesis could be specifically acylated with 3H labeled fatty acids. The order of the lux genes in P. phosphoreum was found to be luxCDABFE with luxF coding for a new polypeptide of 26 kDa. The presence of a new gene in the P. phosphoreum luminescence system between luxB and luxE as compared to the organization of the lux structural gene in V. fischeri and Vibrio harveyi (luxCDABE) demonstrates that the luminescent systems in the marine bacteria have significantly diverged. The discovery of the luxF gene provides the basis for elucidating the role of its gene product in the expression of luminescence in different marine bacteria. PMID- 3049576 TI - Membrane-bound beta-lactamase forms in Escherichia coli. AB - Frameshift pseudo-revertants of Escherichia coli RTEM beta-lactamase were obtained by a selection procedure, starting from frameshift mutants at the signal processing site. These pseudo-revertant proteins, which have a totally altered COOH-terminal part of the signal sequence, are attached to the outer face of the inner membrane. The mutant proteins are enzymatically active in vitro and in vivo, and the membrane localization has no deleterious effect on cell growth. We conclude that initiation of transport across the membrane does not require the COOH-terminal part of the signal, but this part of the sequence determines whether the protein is released to the periplasm either with or without cleavage of the signal, or whether the protein remains anchored to the membrane. Mutants with two signals in series were used to show that a truncated signal is not refractory to transport per se. If neither signal contains a functional cleavage site, the protein is at least partially found on the outer face of the inner membrane. If both signals contain functional cleavage sites, both are removed and the protein is released to the periplasm. If only the first signal contains a cleavage site, a longer fusion protein is transported and released. The results presented here show that a pre-beta-lactamase-like protein can fold properly even as a membrane-bound species. PMID- 3049577 TI - Expression in Escherichia coli of full-length and mutant rat brain calbindin D28. Comparison with the purified native protein. AB - Studies of vitamin D-dependent 28-kilodalton calcium binding protein (calbindin D28) have been hindered by difficulties in purifying large amounts of the protein. In order to overcome this problem, we cloned and expressed a full-length rat brain calbindin D28 cDNA. In addition, we isolated and purified to homogeneity, native rat brain calbindin D28. The isolated native protein has an apparent molecular mass of 27 kDa and properties similar to those of the well characterized chicken calbindin D28. It has an acidic isoelectric point (approximately 4.5), a high affinity for calcium, and an amino terminus blocked to Edman degradation. The properties of the native and the recombinant proteins were examined by gel electrophoresis, isoelectric focusing, protein sequencing, amino acid composition analysis, and calcium binding assays. We demonstrated that: (i) the authentic and the full-length recombinant proteins have similar molecular weights and isoelectric points; (ii) the proteins have the same amino acid composition; (iii) the proteins bind calcium in a similar manner; (iv) the absence of a blocking NH2-terminal group in the recombinant protein does not appreciably influence the binding of calcium. To further examine the calcium binding properties of this protein, we constructed deletion mutants lacking one or both of the two putative degenerated calcium binding sites (EF hand regions). These deletions resulted in smaller proteins that still bound calcium. The ability to express and purify calbindin D28 and mutants thereof should allow the systematic elucidation of structure-function relationships in this class of calcium binding proteins. PMID- 3049578 TI - A noncleavable signal for mitochondrial import of 3-oxoacyl-CoA thiolase. AB - Rat 3-oxoacyl-CoA thiolase, an enzyme of the fatty acid beta-oxidation cycle, is located in the mitochondrial matrix. Unlike most mitochondrial matrix proteins, the thiolase is synthesized with no transient presequence and possesses information for mitochondrial targeting and import in the mature protein of 397 amino acid residues. cDNA sequences encoding various portions of the thiolase were fused in frame to the cDNA encoding the mature portion of rat ornithine transcarbamylase (lacking its own presequence). The fusion genes were transfected into COS cells, and subcellular localization of the fusion proteins was analyzed by cell fractionation with digitonin. When the mature portion of ornithine transcarbamylase was expressed, it was recovered in the soluble fraction. On the other hand, the fusion proteins containing the NH2-terminal 392, 161, or 61 amino acid residues of the thiolase were recovered in the particulate fraction, whereas the fusion protein containing the COOH-terminal 331 residues (residues 62-392) was recovered in the soluble fraction. Enzyme immunocytochemical and immunoelectron microscopic analyses using an anti-ornithine transcarbamylase antibody showed mitochondrial localization of the fusion proteins containing the NH2-terminal portions of the thiolase. These results indicate that the NH2 terminal 61 amino acids of rat 3-oxoacyl-CoA thiolase function as a noncleavable signal for mitochondrial targeting and import of this enzyme protein. Pulse-chase experiments showed that the ornithine transcarbamylase precursor and the thiolase traveled from the cytosol to the mitochondria with half-lives of less than 5 min, whereas the three fusion proteins traveled with half-lives of 10-15 min. Interestingly, in the cells expressing the fusion proteins, the mitochondria showed abnormal shapes and were filled with immunogold-positive crystalloid structures. PMID- 3049580 TI - Sleep-wake regulation by prostaglandins D2 and E2. PMID- 3049582 TI - Receptor-mediated gene delivery and expression in vivo. AB - A soluble DNA carrier system was used to target a foreign gene specifically to liver in vivo via asialoglycoprotein receptors. The DNA carrier was prepared consisting of a galactose-terminal (asialo-)glycoprotein, asialoorosomucoid (AsOR), covalently linked to poly-L-lysine. The conjugate was complexed in a 2:1 molar ratio (based on AsOR content of the conjugate) to the plasmid, pSV2 CAT, containing the gene for the bacterial enzyme chloramphenicol acetyltransferase (CAT). Intravenous injection of [32P]plasmid DNA complexed to the carrier demonstrated specific hepatic targeting with 85% of the injected counts taken up by the liver in 10 min compared to only 17% of the counts when the same amount of [32P]DNA alone was injected under identical conditions. Targeted pSV2 CAT DNA was detected at a level of 1.0 ng/g liver by hybridization of a [32P]pSV2 CAT cDNA probe to rat liver DNA extracted 24 h after intravenous injection of AsOR-poly-L lysine-DNA complex containing 1.0 mg of DNA. Homogenates of livers taken 24 h after injection of the complex revealed that the targeted CAT gene was functional as reflected by the detection of CAT activity (approximately 4 microunits/mg protein). Livers from control animals that received individual constituents of the complex produced no CAT activity. Simultaneous injection of excess AsOR to compete with the AsOR-poly-L-lysine-DNA complex for uptake by the liver inhibited CAT gene expression. Assays for CAT activity in other organs (spleen, kidney, lungs) failed to demonstrate any activity in these organs. This new soluble DNA carrier system can permit targeted delivery of foreign genes specifically to liver with resultant foreign gene expression in vivo. PMID- 3049579 TI - sn-glycerol-3-phosphate acyltransferase tubule formation is dependent upon heat shock proteins (htpR). AB - Overexpression of the Escherichia coli sn-glycerol-3-phosphate (glycerol-P) acyltransferase, an integral membrane protein, causes formation of ordered arrays of the enzyme in vitro. The formation of these tubular structures did not occur in an E. coli strain bearing a mutation in the htpR gene, the regulatory gene for the heat shock response. The htpR165 mutation was shown by genetic analysis to be the lesion responsible for blockage of tubule formation. Similar amounts of glycerol-P acyltransferase were produced in isogenic htpR+ and htpR165 strains, ruling out an effect of htpR165 on expression of glycerol-P acyltransferase. Further, phospholipid metabolism was not altered in either strain after induction of glycerol-P acyltransferase synthesis. Increased glycerol-P acyltransferase synthesis caused a partial induction of the heat shock response which was dependent upon a wild type htpR gene. The heat shock proteins induced were identified as the groEL and dnaK gene products on two-dimensional gels. These two proteins have been implicated in the assembly of bacteriophage coats. These heat shock proteins appear essential for tubule formation. PMID- 3049581 TI - trans-splicing of transcripts for the chloroplast psaA1 gene. In vivo requirement for nuclear gene products. AB - Photosystem I-deficient Chlamydomonas reinhardtii mutants M18 and F27 lack mature mRNA for the psaA1 gene of photosystem I but accumulate unspliced and partially spliced psaA1 transcripts. Analysis of these transcripts indicate that 1) exon 1 of psaA1 is transcribed separately from exon 2, 2) exon 2 is cotranscribed with the psbD gene, and 3) at least two nuclear gene products are required for exon 1 exon 2 trans-splicing. In addition, analysis of chloroplast transcription in permeabilized cells shows that exon 3 is near the 5' end of a separate transcription unit, indicating that exon 2-exon 3 ligation also occurs by a trans splicing mechanism. PMID- 3049583 TI - Isolation and characterization of a small catalytic domain released from the adenylate cyclase from Escherichia coli by digestion with trypsin. AB - An expression plasmid containing a hybrid gene encoding a protein having the primary amino acid sequence of the adenylate cyclase from Escherichia coli was constructed. When the gene was induced, the adenylate cyclase could be expressed at high levels in a cya- strain of E. coli. The majority of the enzymatic activity and protein (having a molecular weight of 95,000) induced was insoluble. However, treatment of the insoluble fraction of cell lysates with trypsin resulted in both an increase in and solubilization of the total amount of adenylate cyclase activity. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the soluble protein produced by treatment with trypsin revealed a polypeptide having a molecular weight of 30,000. This soluble, catalytically active fragment of adenylate cyclase was purified and subjected to amino-terminal sequence analyses; two amino-terminal sequences were identified beginning at residue 82 and at residue 342 of the intact enzyme. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of the purified fragment followed by either silver or Coomassie Blue staining revealed the presence of only a single polypeptide having a molecular weight of 30,000; a short oligopeptide associated with the amino terminus at residue 342 could not be detected. Site-directed mutagenesis was used to place a stop codon at residue 341; the truncated enzyme was catalytically active, so the short oligopeptide is not necessary for catalysis. The Km for ATP, the Ka for Mg2+, and the Vmax determined for the product containing the 30,000-dalton fragment were similar to the values reported for the intact enzyme from E. coli. PMID- 3049584 TI - Properties of Escherichia coli mutants lacking membrane-derived oligosaccharides. AB - Membrane-derived oligosaccharides (MDO) consist of branched substituted beta glucan chains and are present in the periplasmic space of Escherichia coli and other gram-negative bacteria. A procedure for the isolation of mutants defective in MDO synthesis is described. Their phenotype was compared with a mdoA mutant previously identified, and they are mapped in the mdoA region. Mutants lacking MDO showed imparied chemotaxis on tryptone swarm plates, a reduced number of flagella, and an enhanced expression of the OmpC porin. Revertants able to form swarm rings again had regained the ability to synthesize MDO and showed the wild type porin pattern. A second group of chemotactic revertants were mutated in the ompB gene region involved in osmoregulation, and they were still devoid of MDO. These findings provide evidence for a link between MDO biosynthesis and other functions of E. coli related to its adaptation to the environment. PMID- 3049585 TI - DNA sequences of random origin as probes of Escherichia coli promoter architecture. AB - In order to better understand the role of the -35 sequence motif in transcription initiation by Escherichia coli sigma 70 RNA polymerase holoenzyme, -35 promoter elements of limited nucleotide composition have been selected from random DNA sequences. Functional promoter elements have been identified that are composed of just two nucleotide species (A,T; G,C; T,C; and T,G) and one nucleotide species (poly(dT)). From this study of 81 promoter mutations, two conclusions can be made. First, given a population of random DNA sequences, there exist sequences that form stronger -35 promoter elements than the consensus sequence. These sequences lack some features of the consensus sequence and reveal unexpected homology in the ordinarily nonconserved spacer sequence. Second, the sequence requirements at the -35 site may suggest a possible role for DNA secondary structure in the recognition of promoters by RNA polymerase. PMID- 3049586 TI - Glycoprotein biosynthesis in Saccharomyces cerevisiae. Purification of the alpha mannosidase which removes one specific mannose residue from Man9GlcNAc. AB - A soluble form of the specific alpha-mannosidase from Saccharomyces cerevisiae, which catalyzes the following reaction, was purified at least 100,000-fold by conventional chromatography procedures: (Formula: see text). The purified enzyme migrates on sodium dodecyl sulfate-polyacrylamide gel electrophoresis as a single band of about 60 kDa in the absence of reducing agent, and as two bands of about 44.5 kDa and 22.5 kDa in the presence of reducing agent. The apparent molecular weight of the soluble enzyme is about 75,000 by gel filtration on Sephacryl S 200. The specific alpha-mannosidase does not require the addition of divalent cation for activity, but it is inhibited by Tris, EDTA, Mn2+, Co2+, Zn2+, and Mg2+. The inhibition caused by EDTA can be reversed completely by Ca2+ and partially by Mg2+, but not by other divalent cations. The soluble alpha mannosidase arises from a larger hydrophobic form of the enzyme which is found in the detergent phase during partition in Triton X-114. The formation of the soluble enzyme, which is recovered in the aqueous phase during partition in Triton X-114, is time- and temperature-dependent and is prevented by pepstatin, but not by other protease inhibitors. These results indicate that the purified soluble alpha-mannosidase represents the catalytically active domain of the enzyme which has been proteolytically released from its membrane-bound form. PMID- 3049587 TI - Requirements of both glucocorticoids and glucagon as co-inducers for activation of transcription of the serine dehydratase gene in cultured rat hepatocytes. AB - Induction of translatable mRNA for serine dehydratase (SDH) in primary cultured rat hepatocytes requires both dexamethasone and glucagon or cAMP (Noda, C., Tomomura, M., Nakamura, T., and Ichihara, A. (1986) J. Biochem. (Tokyo) 95, 37 45). This unique hormone requirement was studied further with a cDNA probe complementary to SDH mRNA in primary cultured hepatocytes of adult rats. Dot-blot hybridization analysis of RNA showed that SDH mRNA was induced by dexamethasone and glucagon together, but not by either alone. Insulin or epinephrine caused 40% inhibition of this induction of SDH mRNA. Cycloheximide prevented the induction of SDH mRNA by dexamethasone and glucagon, suggesting that ongoing protein synthesis is required for the induction by glucocorticoids and glucagon. In vitro transcription experiments using nuclei isolated from cultured hepatocytes showed that transcription of the SDH gene was not affected by either dexamethasone or glucagon alone, but markedly enhanced by both hormones together, and that this enhancement was inhibited by insulin or epinephrine. These results indicate that the inhibition of SDH induction by epinephrine or insulin was due to effects of these hormones on the transcriptional rate of the SDH gene. PMID- 3049589 TI - Mutagenesis by transient misalignment. AB - Based upon a consideration of two mutational hot spots produced during DNA synthesis by a eukaryotic DNA repair polymerase, we suggested that certain base substitution errors result not from direct miscoding but from correct coding by a transiently misaligned template-primer (Kunkel, T. A., and Alexander, P. S. (1986) J. Biol. Chem. 261, 160-166). This model, which we called dislocation mutagenesis, has been directly tested. Introducing a single, phenotypically silent G----A base change into the template switches the base substitution specificity at the immediately adjacent hot spot, a T residue, from T----G transversions to T----A transversions. The cumulative change in frequency, represented by the disappearance of the T----G events and the appearance of the T ---A events, is greater than 300-fold. These data demonstrate that during DNA synthesis in vitro, a base at one position can code a mutation at another position. This mechanism can operate over greater distances to produce complex mutations as well. We present one example in which a 123-base deletion containing three base changes at one end of the deletion can be precisely explained by transient misalignment. It remains to be established whether mutagenesis by dislocation operates in vivo to produce biologically significant changes in genetic information. PMID- 3049588 TI - Characterization of the cyn operon in Escherichia coli K12. AB - Escherichia coli can overcome the toxicity of environmental cyanate by hydrolysis of cyanate to ammonia and bicarbonate. This reaction is catalyzed by the enzyme cyanase, encoded by the cynS gene. The nucleotide sequence of cynS has been reported (Sung, Y.-c., Anderson, P. M., and Fuchs, J. A. (1987) J. Bacteriol. 169, 5224-5230). The nucleotide sequence of the complete cyn operon has now been determined. The cyn operon is approximately 2600 base pairs and includes cynT, cynS, and cynX, which encode cyanate permease, cyanase, and a protein of unknown function, respectively. Two cyanate-inducible transcripts of 1500 and 2500 nucleotides, respectively, were detected by Northern blot analysis. S1 nuclease mapping experiments indicated that two different cyn mRNAs have a common 5'-end and two different 3'-ends. One 3'-end was located within the coding region of cynX, whereas the other 3'-end includes the entire DNA sequence of cynX. The longer transcript contained 98 nucleotides complementary to lac mRNA produced by the predominant lac transcription termination sequence. Termination vectors were used to show that both 3'-ends were generated by sequences that caused transcriptional termination in vivo. Expression vectors were used to demonstrate that a protein corresponding to the expected size was synthesized from the DNA fragment containing the open reading frame designated cynX. The predicted amino acid sequence of cynX indicates that it is a very hydrophobic protein. The level of cynX expression was significantly less than that of cynT or cynS expression. PMID- 3049590 TI - Retardation of folding as a possible means of suppression of a mutation in the leader sequence of an exported protein. AB - We have proposed (Randall, L. L., and Hardy, S. J. S. (1986) Cell 46, 921-928) that during export of protein from Escherichia coli, there is a kinetic partitioning between the pathway that leads to productive translocation and the pathway that leads to folding of precursors into a stable conformation that is incompatible with export. This model predicts that a decrease in rate along the productive pathway resulting from a defect in the leader sequence could be partially overcome by slowing the folding of the precursor and thereby increasing the time during which that polypeptide would be competent to enter the export pathway. Here it is shown that a change in the mature portion of maltose-binding protein that is known to suppress a mutation in the leader sequence (Cover, W. H., Ryan, J. P., Bassford, P. J., Jr., Walsh, K. A., Bollinger, J., and Randall, L. L. (1987) J. Bacteriol. 169, 1794-1800) also decreases the rate of folding of the precursor. PMID- 3049591 TI - Induction of the manganese-containing superoxide dismutase in Escherichia coli is independent of the oxidative stress (oxyR-controlled) regulon. AB - The synthesis of manganese-superoxide dismutase in response to hydrogen peroxide and to paraquat was examined in strains of Escherichia coli with different mutations in the oxyR gene. Hydrogen peroxide treatment did not induce manganese superoxide dismutase, but did induce the oxyR regulon. Paraquat induced this enzyme in a strain compromised in its ability to induce the defense response against oxidative stress (oxyR deletion) as well as in a strain that is constitutive and overexpresses the oxyR regulon. Catalase (HPI), but not manganese-superoxide dismutase, was over-expressed under anaerobic conditions in a strain harboring a constitutive oxyR mutation. The data clearly demonstrate that the induction of manganese-superoxide dismutase is independent of the oxyR controlled regulon. PMID- 3049592 TI - Structure of a bacterial sensory receptor. A site-directed sulfhydryl study. AB - Cysteines are substituted at six positions in the aspartate receptor, and these mutant proteins are used to investigate three major facets of receptor structure. 1) The surface of the receptor is examined through measurement of the rate constants for chemical modification of the cysteines by aqueous reagents. Different positions exhibit a range of accessibility (for example, Cys-128 most exposed, Cys-36 most buried). 2) The transmembrane structure of the receptor is determined by reaction of the cysteines with a membrane-impermeant reagent. 3) The spatial proximities in the folded structure of specific pairs of cysteines are investigated by disulfide bond formation. These studies illustrate the usefulness of site-directed sulfhydryl chemistry in the analysis of protein structure. PMID- 3049593 TI - Purification and properties of lipid A disaccharide synthase of Escherichia coli. AB - Lipid A disaccharide synthase of Escherichia coli catalyzes the reaction 2,3 diacyl-GlcN-1-P + UDP-2,3-diacyl-GlcN----2',3'-diacyl-GlcN (beta,1'----6)2,3 diacyl-GlcN-1-P + UDP (Ray, B. L., Painter, G., and Raetz, C. R. H. (1984) J. Biol. Chem. 259, 4852-4859). Using a strain that overproduces the enzyme about 200-fold we have devised a simple purification to near homogeneity, utilizing two types of dye-ligand resins and heparin-agarose. The overall purification starting with membrane-free extracts was 54-fold (16,000-fold relative to wild-type extracts) with a 31% yield. The subunit molecular mass determined by sodium dodecyl sulfate gel electrophoresis is approximately 42,000 daltons, and the native enzyme appears to be a dimer. The amino-terminal sequence is (X)-(Thr)-Glu Gln-(X)-Pro-Leu-Thr-Ie-Ala..., consistent with the results predicted from the DNA sequence, Met-Thr-Glu-Gln-Arg-Pro-Leu-Thr-Ile-Ala.... The purified enzyme displays a strong kinetic preference for sugar substrates bearing two fatty acyl moieties, but it is, nevertheless, very useful for the semisynthetic preparation of many lipid A analogs. Gel filtration studies demonstrate that the natural substrates (2,3-diacyl-GlcN-1-P and UDP-2,3-diacyl-GlcN) form micelles (n approximately equal to 300), rather than bilayers, under conditions used to assay the enzyme. Unlike most enzymes of glycerophospholipid synthesis, the lipid A disaccharide synthase does not require the presence of a detergent for catalytic activity. At 1 mM UDP-2,3-diacyl-GlcN the Vmax and Km values for 2,3-diacyl-GlcN 1-P are 14,028 +/- 513 nmol/min/mg and 0.27 +/- 0.02 mM. When 2,3-diacyl-GlcN-1-P is maintained at 1 mM, they are 12,368 +/- 472 nmol/min/mg and 0.11 +/- 0.01 mM for UDP-2,3-diacyl-GlcN. PMID- 3049594 TI - Bovine factor VII. Its purification and complete amino acid sequence. AB - A modified method for purification of blood clotting factor VII from bovine plasma was developed, and its complete amino acid sequence was established. The isolated factor VII was activated with factor XIIa, and the resulting two-chain factor VII (factor VIIa) was reduced and S-pyridylethylated or S-aminoethylated. The amino acid sequences of the S-alkylated heavy and light chains were determined by sequencing the fragments obtained from enzymatic and chemical cleavages. Fast atom bombardment mass spectrometry was also used to establish the COOH-terminal sequence of the heavy chain. The light chain consists of 152 residues with one carbohydrate chain at Asn145, and 11 gamma-carboxyglutamic acid residues are found within the NH2-terminal 35 residues. The light chain contains 0.2-0.3 mol of beta-hydroxyaspartic acid/mol of protein, indicating that an aspartic acid residue in bovine factor VII is incompletely hydroxylated. Moreover, a pentapeptide, Ala-Ser*-Ser-Pro-Cys (positions 51-55), isolated from an enzymatic digest of the light chain, contained an unknown serine derivative, but its structure is still unclear. On the other hand, the heavy chain is composed of 255 residues and one asparagine-linked carbohydrate chain at Asn203. Bovine factor VII, with a total of 407 residues, has 71% sequence identity with the human molecule (406 residues) predicted from the cDNA sequence (Hagen, F. S., Gray, C. L., O'Hara, P., Grant, F. J., Saari, G. C., Woodbury, R. G., Hart, C. E., Insley, M., Kisiel, W., Kurachi, K., and Davie, E. W. (1986) Proc. Natl. Acad. Sci. U. S. A. 83, 2412-2416). PMID- 3049595 TI - Proline porter II is activated by a hyperosmotic shift in both whole cells and membrane vesicles of Escherichia coli K12. AB - Proline porter II is rapidly activated when nongrowing bacteria are subjected to a hyperosmotic shift (Grothe, S., Krogsrud, R. L., McClellan, D. J., Milner, J. L., and Wood, J. M. (1986) J. Bacteriol. 166, 253-259). Proline porter II was active in membrane vesicles prepared from bacteria grown under optimal conditions, nutritional stress, or osmotic stress. That activity was: (i) dependent on the presence of the energy sources phenazine methosulphate plus ascorbate or D-lactate; (ii) observed only when a hyperosmotic shift accompanied the transport measurement; (iii) inhibited by glycine betaine in a manner analogous to that observed in whole cells; and (iv) eliminated by lesions in proP. Membrane vesicles were able to transport serine but not glutamine and serine transport was reduced by the hyperosmotic shift. In whole cells, proline porter II activity was supported by glucose and by D-lactate in a strain defective for proline porters I and III and the F1F0-ATPase. Glucose energized proline uptake was eliminated by carbonyl cyanide m-chlorophenylhydrazone and KCN as was serine uptake. These results suggested that proline porter II was respiration-dependent and probably ion-linked. Activation of proline porter II in whole cells by sucrose or NaCl was sustained over 30 min, whereas activation by glycerol was transient. Proline porter II was activated by NaCl and sucrose with a half-time of approximately 1 min in both whole cells and membrane vesicles. Thus, activation of proline porter II was reversible. It occurred at a rate comparable to that of K+ influx and much more rapid than the genetic regulatory responses that follow a hyperosmotic shift. PMID- 3049596 TI - In addition to RNase H(70) two other proteins of Saccharomyces cerevisiae exhibit ribonuclease H activity. AB - Two ribonucleases H (RNases H) were purified to apparent homogeneity from the yeast Saccharomyces cerevisiae. The enzymes were separated from the previously described yeast ribonuclease H (RNase H(70), Karwan, R., Blutsch, H., and Wintersberger, U. (1983) Biochemistry 22, 5500-5507) by chromatography on Mono Q and blue-Sepharose columns and from each other on a Mono S column. The two proteins, RNase H(55) of molecular weight around 55,000 and RNase H(42) of molecular weight around 42,000, exhibit distinct enzymatic properties: RNase H(55) acts as a 5'-exonuclease of low specific activity and produces predominantly monoribonucleotides from the synthetic hybrid poly(rA)-poly(dT). RNase H(42) efficiently releases oligoribonucleotides from the same substrate. Polyclonal antibodies against these proteins do not cross-react with RNase H(70), and thus, these two RNases H probably do not represent proteolytic breakdown products of RNase H(70). Peptide maps obtained by total digestion of RNase H(55) and RNase H(42) with trypsin reveal several common peptides and, therefore, suggest that the two enzymes are related to each other. We tentatively conclude that RNase H(55) is proteolytically processed to RNase H(42) in vivo. PMID- 3049597 TI - The chlorophyll a/b-binding protein inserts into the thylakoids independent of its cognate transit peptide. AB - In order to determine if the cognate transit peptide of the light-harvesting chlorophyll a/b-binding protein (LHCP) is essential for LHCP import into the chloroplast and proper localization to the thylakoids, it was replaced with the transit peptide of the small subunit (S) of ribulose-1,5-bisphosphate carboxylase/oxygenase, a stromal protein. Wheat LHCP and S genes were fused to make a chimeric gene coding for the hybrid precursor, which was synthesized in vitro and incubated with purified pea chloroplasts. My results show that LHCP is translocated into chloroplasts by the S transit peptide. The hybrid precursor was processed; and most importantly, mature LHCP did not remain in the stroma, but was inserted into thylakoid membranes, where it normally functions. Density gradient centrifugation showed no LHCP in the envelope fraction. Hence, the transit peptide of LHCP is not required for intraorganellar routing, and LHCP itself contains an internal signal for localization to the correct membrane compartment. PMID- 3049598 TI - What is the role of the transit peptide in thylakoid integration of the light harvesting chlorophyll a/b protein? AB - Whereas it is widely accepted that the transit peptide of the precursor for the light-harvesting chlorophyll a/b protein (preLHCP) is responsible for targeting this polypeptide to chloroplasts, the signals which govern its intraorganellar targeting appears to be transit peptide-mediated for plastocyanin (Smeekins, S., Bauerle, C., Hageman, J., Keegstra, K., and Weisbeek, P. (1986) Cell 46, 365-375) and several other nuclear-encoded, thylakoid luminal proteins. To determine whether a similar mechanism operates for LHCP (an integral thylakoid protein), we have used oligonucleotide-directed mutagenesis to delete the proposed transit sequence from a petunia precursor of this polypeptide. Intact preLHCP and the deletion mutant product have been expressed in vitro, and their abilities to integrate into purified thylakoids have been compared. We have found that both polypeptides insert into thylakoids correctly, provided the latter are supplemented with a membrane-free stromal extract and Mg.ATP. Our results clearly demonstrate that whereas the transit peptide is required for transport into chloroplasts, thylakoid integration of preLHCP is determined by mature portions of the polypeptide. In addition, we note that transit peptide removal has little effect on the apparent solubility of the in vitro translation products. PMID- 3049599 TI - Relationship of effective molecular size to systemic clearance in rats of recombinant interleukin-2 chemically modified with water-soluble polymers. AB - We have examined the effects of a variety of chemical modifications to recombinant human interleukin-2 (rIL-2) on its pharmacokinetic behavior in rats. Unmodified rIL-2 is cleared from plasma with half-lives of 3 and 44 min for the alpha and beta phases. Modification of rIL-2 with monomethoxy polyethylene glycol or polyoxyethylated glycerol increased the half-lives as much as 20-fold, although the volume of distribution remained unchanged at 88 +/- 13 ml/kg. The clearance rates correlated with the effective molecular size of the modified protein determined by size exclusion chromatography. Clearance decreased rapidly as the effective molecular size increased from 19.5 to 70 kDa, whereas above 70 kDa the clearance decreased more slowly. This abrupt change at 70 kDa may be related to the permeability threshold of the kidney glomerular membrane which retains proteins larger than albumin in the plasma. Using the relationship between clearance and effective molecular size, the clearance rates of mixtures of modified rIL-2 could be predicted based on their average effective molecular size. Since the effectiveness of rIL-2 therapy is likely to be related to its pharmacokinetic behavior, the ability to design a molecule with a predictable time course in plasma provides a means to study this relationship. PMID- 3049600 TI - Chloroplast transport of a ribulose bisphosphate carboxylase small subunit-5 enolpyruvyl 3-phosphoshikimate synthase chimeric protein requires part of the mature small subunit in addition to the transit peptide. AB - Ribulose bisphosphate carboxylase small subunit protein is synthesized in the cytoplasm as a precursor and transported into the chloroplast where the amino terminal portion, the transit peptide, is removed proteolytically. To obtain chloroplast delivery of the 43-kDa 5-enolpyruvyl 3-phosphoshikimate (EPSP) synthase of Salmonella typhimurium, we constructed fusion proteins between the bacterial EPSP synthase and the ribulose bisphosphate carboxylase small subunit. A fusion protein consisting of the transit peptide fused to the EPSP synthase was not transported in vitro or in vivo into chloroplasts. A second fusion protein consisting of the transit peptide and 24 amino acids of the mature small subunit fused to the EPSP synthase was transported both in vitro and in vivo into chloroplasts. It was processed into two polypeptides of 46 and 47 kDa, respectively. This heterogeneity in processing was not caused by the presence of the aroA start codon, since its removal resulted in the same pattern. Substituting 24 different amino acids for the 24 amino acids of the mature small subunit resulted in a fusion protein that was not transported into the chloroplast. It was concluded that a portion of the mature small subunit was needed for efficient chloroplast delivery. PMID- 3049601 TI - In vivo and in vitro structural analysis of the rplJ mRNA leader of Escherichia coli. Protection by bound L10-L7/L12. AB - The rplJL-rpoBC operon of Escherichia coli is regulated in part at the level of translation by an autogenous mechanism (feedback regulation) that involves ribosomal protein L10-L7/L12. Feedback regulation occurs as the result of L10 L7/L12 binding to a site on the untranslated leader region of the rplJ mRNA that is located more than 100 nucleotides upstream from the translation start site. Previous studies have indicated that the secondary structure of the rplJ leader region is important for efficient translation and feedback regulation. We have done chemical modification experiments to examine the secondary structure of approximately 200 nucleotides of the rplJ leader region, and we propose a secondary structure that is consistent with the experimental data. RNA structure was probed in vitro by treating samples of total cellular RNA with diethyl pyrocarbonate and in vivo by treating log-phase cultures with dimethyl sulfate. Modified bases were detected by primer extension using three different oligonucleotide primers. The proposed structure includes five double-stranded regions designated I to V, separated by single-stranded segments numbered 1 to 5. We have also identified specific nucleotides in the rplJ mRNA leader that are protected by purified L10-L7/L12 from methylation by dimethyl sulfate in vitro. The protected bases are located within a bulge-loop of region IV, a portion of the mRNA that has been shown genetically to be necessary for feedback regulation. PMID- 3049602 TI - Posttranslational processing of alpha-, beta-, and gamma-preprotachykinins. Cell free translation and early posttranslational processing events. AB - We have used an in vitro transcription-translation system to study initial protein processing events of the rat substance P/neurokinin A gene products. cDNA clones for three different mRNA species, which are derived by differential RNA splicing, were subcloned into a plasmid, pGEM1, which contains the promoter for the bacteriophage SP6 RNA polymerase. In vitro synthesized mRNAs for alpha-, beta , and gamma-preprotachykinin were translated in a wheat germ or rabbit reticulocyte cell-free system. When analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis, the translated protein products migrate consistent with the deduced molecular masses of alpha (13,035 Da)-, beta (15,003 Da)-, and gamma (13,343 Da)-preprotachykinin. The addition of dog pancreatic microsomal membranes to either cell-free translation system causes the production of a protease-resistant form of each of the three preprotachykinins which migrates with an apparent increase in molecular mass of approximately 2,000 Da. Each of these modified preprotachykinins lacks the putative signal peptide of the prepro- form, with signal peptidase cleavage occurring after the alanine residue at position 19. Both the prepro- and proforms of each tachykinin precursor molecule are recognized by antiserum R-140, an antiserum specific for the mid portion of the undecapeptide substance P. The most likely explanation for the apparent increase in molecular mass is anomalous electrophoretic migration, since beta-preprotachykinin mRNA lacking the signal peptide encoding sequence is translated, in the absence of microsomal membranes, into a protein with the same apparent molecular mass as the modified form of beta-preprotachykinin. Therefore, each of the three preprotachykinin mRNAs are translatable, and their products are targeted to the secretory pathway by the presence of a cleavable signal peptide. PMID- 3049603 TI - Renaturation of DNA by a Saccharomyces cerevisiae protein that catalyzes homologous pairing and strand exchange. AB - A protein from mitotic Saccharomyces cerevisiae cells that catalyzes homologous pairing and strand exchange was analyzed for the ability to catalyze other related reactions. The protein was capable of renaturing complementary single stranded DNA as evidenced by S1 nuclease assays and analysis of the reaction products by agarose gel electrophoresis and electron microscopy. Incubation of the yeast protein with complementary single-stranded DNA resulted in the rapid formation of large aggregates which did not enter agarose gels. These aggregates contained many branched structures consisting of both single-stranded and double stranded DNA. These reactions required stoichiometric amounts of protein but showed no ATP requirement. The protein formed stable complexes with both single stranded and double-stranded DNA, showing a higher affinity for single-stranded DNA. The binding to single-stranded DNA resulted in the formation of large protein:DNA aggregates. These aggregates were also formed in strand-exchange reactions and contained both substrate and product DNAs. These results demonstrate that the S. cerevisiae strand-exchange protein shares additional properties with the Escherichia coli recA protein which, by analogy, gives further indication that it might be implicated in homologous recombination. PMID- 3049605 TI - Nonlinear relationship between concentration and activity of a bacterial ice nucleation protein. AB - The expression level of an ice nucleation gene (inaZ) was varied in Escherichia coli to observe the relationship between activity and gene product. The ice nucleation activity increased as the 2nd to 3rd power of the membrane concentration of the inaZ gene product, implying that molecules of InaZ protein interact cooperatively in groups of two to three at the rate-limiting step of ice nucleus assembly. The 2nd to 3rd power relationship was independent of the threshold temperature at which ice nucleation was measured and was consistent over a 500-fold range of protein concentration. Such a relationship indicates that the same rate-limiting step must be common to the formation of ice nuclei displaying all the various threshold temperatures within a bacterial population. Observations of Pseudomonas syringae, expressing the inaZ gene at various levels, were consistent with a similar relationship and hence a similar mechanism of ice nucleus assembly in Pseudomonas. PMID- 3049604 TI - Messenger RNA orientation on the ribosome. Placement by electron microscopy of antibody-complementary oligodeoxynucleotide complexes. AB - Messenger RNA orients on the small ribosomal subunit by base pairing with a complementary sequence in ribosomal RNA. We have positioned this ribosomal RNA segment and thus oriented the mRNA using a new technique--localization of an antibody-recognizable modified complementary oligodeoxynucleotide by electron microscopy. A synthetic oligodeoxynucleotide complementary to the message positioning ribosomal RNA sequence was modified at either or both ends with different antigenic markers. Electron microscopy of subunit-oligodeoxynucleotide antibody complexes allowed separate placement of each terminal marker of the oligodeoxynucleotide probe. The 5'-end of the complementary sequence contacts the subunit at the platform tip (rRNA nucleotide 1542). The message then extends along the interior side of the platform to the level of the fork of the cleft separating the platform from the subunit body, and displaced slightly to the convex side of the platform (rRNA nucleotide 1531). Based on our results and data from other laboratories, we propose a model for the positioning of messenger RNA on the 30 S subunit. PMID- 3049606 TI - The two-component, ATP-dependent Clp protease of Escherichia coli. Purification, cloning, and mutational analysis of the ATP-binding component. AB - The ATP-binding component (Component II, hereafter referred to as ClpA) of a two component, ATP-dependent protease from Escherichia coli has been purified to homogeneity. ClpA is a protein with subunit Mr 81,000. It has an intrinsic ATPase activity and activates degradation of protein substrates only in the presence of a second component (Component I, hereafter referred to as ClpP), Mg2+, and ATP. The amount of ClpA varies by less than a factor of 2 in cells grown in different media and at temperatures from 30 to 42 degrees C. ClpA does not appear to be a heat-shock protein since its synthesis is not dependent on htpR. Antibodies against purified ClpA were used to identify lambda transducing phage bearing the clpA gene. The cloned gene contains a DNA sequence expected to code for the first 28 amino acids of ClpA, which were determined by protein sequencing of purified ClpA. The clpA gene in the phage was mutated by insertion of delta kan defective transposons and the mutations were transferred to E. coli by homologous recombination. The clpA gene was mapped to 19 min on the E. coli chromosome. Mutant cells with insertions early in the gene produce no ClpA protein detectable in Western blots, and extracts of such mutant cells have no detectable ClpA activity. clpA- mutants grow well under all conditions tested and are not defective in turnover of proteins during nitrogen starvation nor in the turnover of such highly unstable proteins as the lambda proteins O, N, and cII, or the E. coli proteins SulA, RcsA, and glutamate dehydrogenase. The degradation of abnormal canavanine-containing proteins is defective in clpA mutants especially in cells that also have a lon- mutation. Extracts of clpA- lon- cells have ATP dependent casein degrading activity. PMID- 3049607 TI - Structure and expression of the genes encoding the alpha and beta subunits of yeast phenylalanyl-tRNA synthetase. AB - The two genes FRS1 and FRS2 encoding, respectively, the large (alpha) and small (beta) subunits of cytoplasmic phenylalanyl-tRNA synthetase from bakers' yeast have been cloned and sequenced. The derived protein primary structures are confirmed by peptide sequences evenly distributed along the reading frames. These predict a subunit Mr of 67,347 for alpha and 57,433 for beta, in good agreement with earlier determinations carried out on the purified protein. These subunit sequences have been compared to those of Escherichia coli phenylalanyl-tRNA synthetase as well as to the small beta subunit of the corresponding yeast mitochondrial enzyme; limited but significant homology was found between the two alpha subunits on the one hand and between the three beta subunits on the other hand. The results suggest that these three enzymes, from E. coli, yeast cytoplasm, and yeast mitochondria, have strongly diverged from one another. The initiation sites of transcription have been determined for both yeast genes. Their 5'-upstream regions show no sequence similarities that would have indicated a coordinate control of gene expression at the transcriptional level. Measurements of steady-state levels of FRS-mRNAs in overproducing strains indicate that there is no restriction in mRNA synthesis. Therefore the control of gene expression, leading to a balanced synthesis of alpha and beta subunits, is likely to occur at the translational level. PMID- 3049608 TI - Specificity of mutagenesis by 4-aminobiphenyl. A possible role for N (deoxyadenosin-8-yl)-4-aminobiphenyl as a premutational lesion. AB - Mutagenesis by N-acetoxy-N-trifluoroacetyl-4-aminobiphenyl, a reactive form of the human bladder carcinogen 4-aminobiphenyl (ABP), was studied in Escherichia coli virus M13mp10. N-acetoxy-N-trifluoroacetyl-4-ABP-treated DNA containing 140 lesions/duplex genome, when introduced into excision repair-competent cells induced for SOS mutagenic processing, resulted in a 40-fold increase in mutation frequency over background in the lacZ alpha gene fragment. DNA sequence changes were determined for 20 independent mutants. G-C base pairs were the major targets for base pair substitution mutations, although significant mutagenic activity was also observed at certain A-T base pairs. Deletion and frameshift mutations also were found in this sample. The salient feature of this partial "mutational spectrum" was a hotspot that occurred at position 6357 (amino acid 30 of the beta galactosidase fragment encoded by M13mp10); this A-T to T-A transversion appeared in 6 of the 20 mutants. The property of ABP to mutate A-T base pairs was consistent with the result that N-hydroxy-ABP reverted Salmonella typhimurium strain TA104, which is presumed to revert primarily due to mutations at these sites. The ability of the major carcinogen-DNA adduct formed by ABP in vivo and in vitro, N-(deoxyguanosin-8-yl)-4-aminobiphenyl, to cause base pair substitution mutations was also investigated. This adduct was positioned specifically in the minus strand at position 6270 in duplex M13mp10 DNA. In the presence of the mutagenesis-enhancing plasmid pGW16 and UV induction of SOS mutagenic processing, it was shown that fewer than 0.02% of the adducts resulted in transition or transversion mutations following transfection of DNA into excision-repair competent cells. Similar results were obtained in uvrA and uvrC backgrounds. Although the major adduct did not cause base substitution mutations under these experimental conditions, the contribution of this lesion to the entire spectrum of mutations in the lacZ alpha fragment seems likely. PMID- 3049609 TI - The intrinsic fluorescence of the alpha subunit of transducin. Measurement of receptor-dependent guanine nucleotide exchange. AB - We have made use of the enhancement of the intrinsic fluorescence of the alpha subunit of transducin (alpha T), which accompanies guanine nucleotide exchange, to follow the reconstituted interactions between pure rhodopsin and pure transducin in phospholipid vesicles. When the pure alpha T.GDP complex is added to lipid vesicles containing rhodopsin and the beta gamma T complex, a light- and guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S)-dependent enhancement of the fluorescence emission of alpha T is observed. When GTP is substituted for GTP gamma S, a similar enhancement of the intrinsic fluorescence of alpha T occurs; however, this enhancement is transient and precedes a fluorescence decay which is complete in 2-5 min. The fact that the fluorescence decay is specifically induced by GTP and is not observed either with nonhydrolyzable GTP analogs or with NaF (plus AlCl3) indicates that the decay represents GTP hydrolysis in alpha T. The dose-response profiles for the effects of the beta gamma T complex on the rate and extent of the GTP gamma S-stimulated fluorescence enhancement of alpha T have also been examined. The addition of relatively low levels of beta gamma T to these reconstituted systems can promote the GTP gamma S-stimulated enhancement of the fluorescence of multiple alpha T subunits with half-maximal enhancement occurring at alpha T:beta gamma T ratios of 150:1. These findings are consistent with earlier suggestions (Fung, B. K.-K. (1983) J. Biol. Chem. 258, 10495-10502) that the beta gamma T subunit dissociates from alpha T as a result of the GDP-GTP exchange reaction and thus can act catalytically to promote the activation of a number of inactive alpha T species. However, the dependence of the rate of the GTP gamma S-stimulated fluorescence enhancement on beta gamma T is complex and cannot be explained adequately by simple models where alpha T-beta gamma T interactions (or rhodopsin-transducin interactions) are rate-limiting for the rhodopsin-stimulated activation of the alpha T subunits. Overall, the results reported here demonstrate that fluorescence spectroscopy can be used to monitor directly a receptor-catalyzed activation-deactivation cycle of a GTP-binding protein within a lipid milieu. PMID- 3049610 TI - Identification, purification, and partial characterization of a novel Mr 28,000 integral membrane protein from erythrocytes and renal tubules. AB - A novel Mr 28,000 integral membrane protein ("28kDa") was identified in human erythrocytes and found entirely associated with the Triton X-100 insoluble membrane skeletons. Antibodies to 28kDa reacted strongly on immunoblots with 28kDa and a diffuse region of Mr 35,000-60,000 ("HMW-28kDa"). Selective proteolytic digestions of membranes demonstrated that HMW-28kDa has an extracellular domain, and both 28kDa and HMW-28kDa have intracellular domains. 28kDa and HMW-28kDa were purified to homogeneity. Quantitative immunoblots indicate that each erythrocyte contains 120,000-160,000 copies of 28kDa. Two dimensional iodopeptide maps of 28kDa and HMW-28kDa were nearly identical; peptide-N-glycosidase digestion of purified HMW-28kDa demonstrated that it is the N-glycosylated form of 28kDa. When concentrated, 28kDa formed a series of larger oligomers which were stable in sodium dodecyl sulfate. Of several nonerythroid tissues studied with anti-28kDa immunoblots, only kidney displayed immunoreactive 28kDa. Purified rat kidney 28kDa was nearly identical to rat erythrocyte 28kDa when compared by two-dimensional iodopeptide mapping. Immunohistochemical staining of human kidney with anti-28kDa demonstrated prominent staining over the apical brush borders of proximal convoluted tubules. A novel integral membrane protein has been purified from erythrocyte and kidney membranes. This new protein may play a role in linkage of the membrane skeleton to the lipid bilayer. PMID- 3049611 TI - A Chinese hamster cell cycle mutant arrested at G2 phase has a temperature sensitive ubiquitin-activating enzyme, E1. AB - The effect of restrictive temperature on ubiquitin conjugation activity has been studied in cells of ts20, a temperature-sensitive cell cycle mutant of the Chinese hamster cell line E36. Ts20 is arrested in early G2 phase at nonpermissive temperature. Immunoblotting with antibodies to ubiquitin conjugates shows that conjugates disappear rapidly at restrictive temperatures in ts20 mutant but not in wild type E36 cells. The incorporation of 125I-ubiquitin into permeabilized ts20 cells is temperature-sensitive. Addition of extracts of another G2 phase mutant, FM3A ts85, with a temperature-sensitive ubiquitin activation enzyme (E1), to permeabilized ts20 cells at restrictive temperatures fails to complement their ubiquitin ligation activity. This indicates that the lesions in the two mutants are similar. Purified E1 from reticulocytes restores the conjugation activity of heat-inactivated permeabilized ts20 cells. Ubiquitin conjugation activity of cell-free extracts of ts20 cells was temperature sensitive and could be restored by adding purified reticulocyte E1. Purified reticulocyte E2 or E3, on the other hand, did not restore the ubiquitin conjugation activity of heat-treated ts20 extracts. These results are consistent with the conclusion that ts20 has temperature-sensitive ubiquitin-activating enzyme (E1). The fact that two E1 mutants (ts20 and ts85) derived from different cell lines are arrested at the S/G2 boundary at restrictive temperatures strongly indicates that ubiquitin ligation is necessary for passage through this part of the cell cycle. The temperature thresholds of heat shock protein synthesis of ts20 and wild type E36 cells were identical. The implications of these findings with respect to a suggested role of ubiquitin in coupling between protein denaturation and the heat shock response are discussed. PMID- 3049612 TI - Proteolytic processing of rat liver membrane secretory component. Cleavage activity is localized to bile canalicular membranes. AB - Membrane secretory component (mSC) mediates the transcellular movement of polymeric IgA from the sinusoidal to the bile canalicular surface of rat hepatocytes. Prior to or concomitant with arrival at the bile canalicular membrane, mSC is cleaved, producing a soluble proteolytic fragment (fSC) which is released into the bile. Conversion of mSC to fSC occurs at the cell surface of cultured rat hepatocytes (Musil, L. S., and Baenziger, J. U. (1987) J. Cell Biol. 104, 1725-1733), suggesting that vectorial release of fSC into bile in vivo may reflect localization of a mSC-specific protease to bile canalicular membranes. We have established a reconstituted system to examine the process of specific cleavage of mSC to yield fSC and to characterize the protease activity responsible. A membrane fraction highly enriched for endocytic vesicles was found to contain approximately 90% of the [35S]Cys-mSC from metabolically labeled rat liver slices but only 5% of the cellular protein. No cleavage activity was present in these vesicles. Highly enriched bile canalicular membranes were able to mediate cleavage of metabolically labeled mSC to a fragment indistinguishable from authentic fSC. In the absence of nonionic detergent, cleavage was dependent on the presence of polyethylene glycol, presumably to mediate fusion of mSC enriched membranes with bile canalicular membranes. Following solubilization with nonionic detergent, cleavage was no longer dependent on the addition of polyethylene glycol. Cleavage of mSC was not observed with either intact or detergent-solubilized sinusoidal, microsomal, or lysosomal membranes. We have thus identified a proteolytic activity associated with bile canalicular membranes which has the properties of a membrane protein and is likely to be responsible for production of fSC in vivo. Its highly restricted localization to the bile canalicular membrane would account for the vectorial release of fSC into the bile. PMID- 3049613 TI - Periplasmic nonspecific acid phosphatase II from Salmonella typhimurium LT2. Crystallization, detergent reactivation, and phosphotransferase activity. AB - The periplasmic nonspecific acid phosphatase II from Salmonella typhimurium was purified to homogeneity from a mutant strain that overproduces the enzyme (Uerkvitz, W., and Beck, C.F. (1981) J. Biol. Chem. 256, 382-389). It was shown that the enzyme transfers phosphate groups from organic phosphoric acid esters (donors) to water as well as to the 2'-, 3'-, or 5'-hydroxyls of nucleosides, nucleotides, and other compounds with free hydroxyl groups (acceptors). The enzyme was crystallized in two forms by precipitation with polyethylene glycol. Needles were formed in buffer containing Mg2+, whereas thin rectangular plates appeared in the presence of the non-ionic detergent n-octyl glucoside. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis under partially or completely denaturating conditions revealed that the native enzyme is a tetramer consisting of identical 24-kDa monomers. Owing to surface inactivation, polyethylene glycol, non-ionic, or Zwitterionic detergents are indispensable for enzyme stability. The detergents are able to reactivate inactivated enzyme when present near or above their critical micelle concentration. PMID- 3049614 TI - World Health Organization International Standards for highly purified human, porcine and bovine insulins. AB - The 4th International Standard (IS) for Insulin, established in 1958, consists of a mixture of relatively impure bovine and porcine insulins and is not suitable as a standard for the assay of highly purified single-species insulins presently used in the treatment of diabetes. Preparations of human, bovine and porcine crystalline insulins, representative of current highly purified therapeutic insulins, have now been studied in an international collaborative study carried out by twenty-three laboratories in fifteen countries. In the collaborative study described here, each of the three preparations was found to be suitable for use as a standard for insulin for bioassay and each was established by WHO in 1986 as an international standard. The 4th IS of Insulin bovine/porcine (code numbered 58/6) has been discontinued. Insulin preparations should now be calibrated in terms of International Units defined by the standard for the appropriate species: the International Standard for Insulin, Human, the International Standard for Insulin, Bovine, or the International Standard for Insulin, Porcine. PMID- 3049615 TI - WHO international reference reagents for bovine and porcine proinsulins. AB - Candidate preparations for International Reference Reagents (IRR) for immunoassays of bovine and porcine proinsulin were evaluated in an international collaborative study. With the authorization of the Expert Committee on Biological Standardization of WHO, the following preparations were established as IRRs: bovine proinsulin (code 84/514, defined ampoule content 25 micrograms) and porcine proinsulin (84/528, 20 micrograms). The content of ampoules of these materials is defined in terms of mass rather than international units of activity, therefore they are IRR rather than International Standards. Both preparations are intended as primary reference reagents for the calibration of immunoassays. PMID- 3049616 TI - [Method of titration of Chlamydia psittaci by immunofluorescence]. PMID- 3049618 TI - N-myc proto-oncogene expression during organogenesis in the developing mouse as revealed by in situ hybridization. AB - The N-myc proto-oncogene is expressed during embryogenesis, suggesting that it plays a role in normal development. Since the myc-family oncogenes have been implicated in the control of cell growth, the embryonic expression may reflect rapid proliferation known to occur in development. Alternatively, N-myc expression may be involved in specific differentiation stages. In many embryonic tissues, early and late differentiation events occur in different locations. By in situ hybridization of tissue sections, we now demonstrate a restricted expression of N-myc mRNA to a few tissues and to areas where the first differentiation stages occur. N-myc expression was most strongly expressed in the developing kidney, hair follicles, and in various parts of the central nervous system. In these tissues, expression was restricted to a few cell lineages. In all lineages, expression was confined to early differentiation stages, and, at onset of overt differentiation, the level of expression decreased dramatically. Several rapidly proliferating tissues showed very little, if any, N-myc expression. In the brain, post-mitotic but not yet differentiated cells expressed high levels of N-myc mRNA. Therefore, N-myc expression is not a simple marker for proliferation in the embryo. Rather, N-myc expression seems to be a feature of early differentiation stages of some cell lineages in kidney, brain, and hair follicles, regardless of the proliferative status of the cell. The results raise the possibility that N-myc may participate in the control of these early differentiation events. PMID- 3049617 TI - Cachectin/tumor necrosis factor exerts endocrine, paracrine, and autocrine control of inflammatory responses. PMID- 3049619 TI - Organelle assembly in yeast: characterization of yeast mutants defective in vacuolar biogenesis and protein sorting. AB - Yeast vacuole protein targeting (vpt) mutants exhibit defects in the sorting and processing of multiple vacuolar hydrolases. To evaluate the impact these vpt mutations have on the biogenesis and functioning of the lysosome-like vacuole, we have used light and electron microscopic techniques to analyze the vacuolar morphology in the mutants. These observations have permitted us to assign the vpt mutants to three distinct classes. The class A vpt mutants (26 complementation groups) contain 1-3 large vacuoles that are morphologically indistinguishable from those in the parental strain, suggesting that only a subset of the proteins destined for delivery to this compartment is mislocalized. One class A mutant (vpt13) is very sensitive to low pH and exhibits a defect in vacuole acidification. Consistent with a potential role for vacuolar pH in protein sorting, we found that bafilomycin A1, a specific inhibitor of the vacuolar ATPase, as well as the weak base ammonium acetate and the proton ionophore carbonyl cyanide m-chlorophenylhydrazone, collapse the pH gradient across the vacuolar membrane and cause the missorting and secretion of two vacuolar hydrolases in wild-type cells. Mutants in the three class B vpt complementation groups exhibit a fragmented vacuole morphology. In these mutants, no large normal vacuoles are observed. Instead, many (20-40) smaller vacuole-like organelles accumulate. The class C vpt mutants, which constitute four complementation groups, exhibit extreme defects in vacuole biogenesis. The mutants lack any organelle resembling a normal vacuole but accumulate other organelles including vesicles, multilamellar membrane structures, and Golgi-related structures. Heterozygous class C zygotes reassemble normal vacuoles rapidly, indicating that some of the accumulated aberrant structures may be intermediates in vacuole formation. These class C mutants also exhibit sensitivity to osmotic stress, suggesting an osmoregulatory role for the vacuole. The vpt mutants should provide insights into the normal physiological role of the vacuole, as well as allowing identification of components required for vacuole protein sorting and/or vacuole assembly. PMID- 3049620 TI - Functions of microtubules in the Saccharomyces cerevisiae cell cycle. AB - We used the inhibitor nocodazole in conjunction with immunofluorescence and electron microscopy to investigate microtubule function in the yeast cell cycle. Under appropriate conditions, this drug produced a rapid and essentially complete disassembly of cytoplasmic and intranuclear microtubules, accompanied by a rapid and essentially complete block of cellular and nuclear division. These effects were similar to, but more profound than, the effects of the related drug methyl benzimidazole carbamate (MBC). In the nocodazole-treated cells, the selection of nonrandom budding sites, the formation of chitin rings and rings of 10-nm filaments at those sites, bud emergence, differential bud enlargement, and apical bud growth appeared to proceed normally, and the intracellular distribution of actin was not detectably perturbed. Thus, the cytoplasmic microtubules are apparently not essential for the establishment of cell polarity and the localization of cell-surface growth. In contrast, nocodazole profoundly affected the behavior of the nucleus. Although spindle-pole bodies (SPBs) could duplicate in the absence of microtubules, SPB separation was blocked. Moreover, complete spindles present at the beginning of drug treatment appeared to collapse, drawing the opposed SPBs and associated nuclear envelope close together. Nuclei did not migrate to the mother-bud necks in nocodazole-treated cells, although nuclei that had reached the necks before drug treatment remained there. Moreover, the double SPBs in arrested cells were often not oriented toward the budding sites, in contrast to the situation in normal cells. Thus, microtubules (cytoplasmic, intranuclear, or both) appear to be necessary for the migration and proper orientation of the nucleus, as well as for SPB separation, spindle function, and nuclear division. PMID- 3049621 TI - Truncation of the carboxy-terminal domain of yeast beta-tubulin causes temperature-sensitive growth and hypersensitivity to antimitotic drugs. AB - beta-tubulin of budding yeast Saccharomyces cerevisiae is a polypeptide of 457 amino acids encoded by the unique gene TUB2. We investigated the function of the carboxy-terminal part of yeast beta-tubulin corresponding to the carboxy-terminal variable domain of mammalian and avian beta-tubulins. The GAA codon for Glu-431 of TUB2 was altered to TAA termination codon by using in vitro site-directed mutagenesis so that the 27-amino acid residues of the carboxyl terminus was truncated when expressed. The mutagenized TUB2 gene (tub2(T430)) was introduced into a haploid strain in which the original TUB2 gene had been disrupted. The tub2(T430) haploid strain grows normally less than 30 but not at 37 degrees C. The truncation of the carboxyl terminus caused hypersensitivity to antimitotic drugs and low spore viability at the permissive temperature for vegetative growth. Immunofluorescence labeling with antitubulin antibody and DNA staining with 4',6'-diamidino-2-phenylindole showed that in these cells at 37 degrees C, formation of spindle microtubules and nuclear division was inhibited and cytoplasmic microtubule distribution was aberrant. These results suggest that functions of the carboxy-terminal domain of yeast beta-tubulin are necessary for cells growing under suboptimal growth conditions although it is not essential for growth under the optimal growth conditions. Cells bearing tub2(411), a tub2 gene in which the GAA codon for Glu-412 was altered to TAA were no more viable at any temperature. In addition, a haploid strain carrying two functional beta-tubulin genes is not viable. PMID- 3049623 TI - Functional alterations in beta'-actin from a KB cell mutant resistant to cytochalasin B. AB - We recently described the isolation of mutant KB cells (Cyt 1 cells) resistant to the cytotoxic effect of cytochalasin B (CB). This mutant carried an altered beta actin; i.e., beta'-actin (Toyama, S., and S. Toyama. 1984. Cell. 37:609-614). In the present study, we have examined the functional properties of actin in Cyt 1 cells. Our results showed that increased resistance of Cyt 1 cells to CB was reflected in altered properties of beta'-actin itself. This was shown directly by two findings. First, the polymerization of beta'-actin was more resistant than that of beta- or gamma-actin to the multiple effects of CB. Second, beta'-actin bound less CB than beta- or gamma-actin. The functional alteration of beta'-actin in Cyt 1 cells was further supported by the observation that, although treatment of KB cells with CB increased the pool of unpolymerized actin, the same treatment did not affect the pool of unpolymerized actin in Cyt 1 cells, and that microfilaments of Cyt 1 cells were more resistant to the disrupting action of CB than those of KB cells. These results strongly suggest that the primary site of action of CB on cell motility processes is actin. PMID- 3049622 TI - Reconstitution of protein transport from the endoplasmic reticulum to the Golgi complex in yeast: the acceptor Golgi compartment is defective in the sec23 mutant. AB - Using either permeabilized cells or microsomes we have reconstituted the early events of the yeast secretory pathway in vitro. In the first stage of the reaction approximately 50-70% of the prepro-alpha-factor, synthesized in a yeast translation lysate, is translocated into the endoplasmic reticulum (ER) of permeabilized yeast cells or directly into yeast microsomes. In the second stage of the reaction 48-66% of the ER form of alpha-factor (26,000 D) is then converted to the high molecular weight Golgi form in the presence of ATP, soluble factors and an acceptor membrane fraction; GTP gamma S inhibits this transport reaction. Donor, acceptor, and soluble fractions can be separated in this assay. This has enabled us to determine the defective fraction in sec23, a secretory mutant that blocks ER to Golgi transport in vivo. When fractions were prepared from mutant cells grown at the permissive or restrictive temperature and then assayed in vitro, the acceptor Golgi fraction was found to be defective. PMID- 3049625 TI - The role of the cell adhesion molecule uvomorulin in the formation and maintenance of the epithelial junctional complex. AB - The role of the epithelial adhesion molecule uvomorulin in the formation of the epithelial junctional complex in the Madin-Darby canine kidney (MDCK) cell line was investigated. Experiments were carried out to determine whether specific inhibition of uvomorulin function would interfere selectively with the formation, stability, or function of the apical zonula adherens (ZA) and zonula occludens (ZO), or whether it would interfere with all forms of intercellular contact including the desmosomes. The effects of blocking antibodies and Fab fragments to uvomorulin on the formation of the junctional complex was examined with a Ca2+ switch assay for de novo junction assembly. The formation of the ZO, the ZA, and the desmosomes was assayed by fluorescence staining with an antibody to the tight junction-specific protein ZO-1, with rhodamine-phalloidin for ZA-associated actin filaments, and with an anti-desmoplakin antibody, respectively. Under different conditions and times of antibody treatment the extent of inhibition of the formation of each of the junctional elements was very similar. The ability of the cells to eventually overcome the inhibitory effect of the antibodies and form junctions correlated with the reappearance of uvomorulin at the regions of cell cell contact. Therefore uvomorulin seems to mediate an early adhesion event between epithelial cells that is a prerequisite for the assembly of all elements of the junctional complex. In contrast, the transepithelial electrical resistance of confluent, well-established monolayers of MDCK cells grown on filters was not greatly affected by treatment with the various antibodies or Fab fragments. A small transient decrease in resistance observed with the polyclonal alpha uvomorulin IgG may be due to a more subtle modulation of the junctional complex. PMID- 3049626 TI - Role of laminin and basement membrane in the morphological differentiation of human endothelial cells into capillary-like structures. AB - We have defined a signal responsible for the morphological differentiation of human umbilical vein and human dermal microvascular endothelial cells in vitro. We find that human umbilical vein endothelial cells deprived of growth factors undergo morphological differentiation with tube formation after 6-12 wk, and that human dermal microvascular endothelial cells differentiate after 1 wk of growth factor deprivation. Here, we report that morphological differentiation of both types of endothelial cells is markedly accelerated by culture on a reconstituted gel composed of basement membrane proteins. Under these conditions, tube formation begins in 1-2 h and is complete by 24 h. The tubes are maintained for greater than 2 wk. Little or no proliferation occurs under these conditions, although the cells, when trypsinized and replated on fibronectin-coated tissue culture dishes, resume division. Ultrastructurally, the tubes possess a lumen surrounded by endothelial cells attached to one another by junctional complexes. The cells possess Weibel-Palade bodies and factor VIII-related antigens, and take up acetylated low density lipoproteins. Tubule formation does not occur on tissue culture plastic coated with laminin or collagen IV, either alone or in combination, or on an agarose or a collagen I gel. However, endothelial cells cultured on a collagen I gel supplemented with laminin form tubules, while supplementation with collagen IV induces a lesser degree of tubule formation. Preincubation of endothelial cells with antibodies to laminin prevented tubule formation while antibodies to collagen IV were less inhibitory. Preincubation of endothelial cells with synthetic peptides derived from the laminin B1 chain that bind to the laminin cell surface receptor or incorporation of these peptides into the gel matrix blocked tubule formation, whereas control peptides did not. These observations indicate that endothelial cells can rapidly differentiate on a basement membrane-like matrix and that laminin is the principal factor in inducing this change. PMID- 3049627 TI - Specific binding, internalization, and degradation of human recombinant interleukin-3 by cells of the acute myelogenous, leukemia line, KG-1. AB - We have studied the interaction of 35S-labeled recombinant IL-3 with the acute myelogenous leukemia cell line, KG-1. 35S-IL-3 bound to these cells in a time dependent, saturable, and specific manner at 4 degrees C. Scatchard transformation of binding isotherms demonstrated the existence of a small number (200) of binding sites, with an apparent dissociation constant of 70-105 pM. After a temperature shift from 4 degrees C to 37 degrees C, surface-bound 35S-IL 3 was rapidly internalized and processed into a trichloroacetic acid soluble form that was released into the medium. Experiments to address the specificity of the IL-3 binding site revealed that neither human IL-2, M-CSF, erythropoietin, transferrin, bovine insulin, nor murine nerve growth factor compete with IL-3 for binding to KG-1 cells. Both human and gibbon recombinant IL-3 and, surprisingly, human recombinant GM-CSF effectively competed the binding of the labeled IL-3 to these cells at 4 degrees C. The competition by GM-CSF was found to be concentration dependent, but much higher concentrations were required to achieve the levels obtained with IL-3. These results suggest that GM-CSF may also interact with the high-affinity IL-3 binding site on KG-1 cells or, alternatively, that GM-CSF binding to its own receptor may decrease the affinity of the IL-3 receptor for its ligand. PMID- 3049628 TI - Intercellular adhesion molecule-1 (ICAM-1) is involved in the cytolytic T lymphocyte interaction with a human synovial cell line. AB - The cell surface molecules involved in the human cytolytic T lymphocyte (CTL) synovial cell interaction may play an important role in T cell interactions with connective tissue mesenchymal cells. To examine the molecular basis for the CTL synovial cell interaction, we immortalized synovial cell explants to establish the cell line SYN.SPP. The SYN.SPP cell line was compared to the established B lymphoblastoid cell line JY. Cell surface immunofluorescence demonstrated significantly different levels of the immunologically relevant cell surface molecules ICAM-1 and LFA-3. Both cell lines were used to stimulate CTL precursors. After several months in culture, CTL lines stimulated by the SYN.SPP and JY cell lines demonstrated HLA class I-directed cytolytic activity. The cell surface molecules utilized by the anti-SYN.SPP and anti-JY CTL lines were identified by monoclonal antibody (MAb) inhibition. MAb recognizing the CTL cell surface molecules CD3, CD8 and LFA-1 (CD11a) significantly inhibited CTL-mediated lysis of both target cells. An interesting observation was that the anti-SYN.SPP CTL line appeared to utilize the ICAM-1 and not the LFA-3 target cell molecule. In contrast, the anti-JY CTL line utilized the LFA-3 and not the ICAM-1 membrane molecule. These results indicate that CTL interactions with connective tissue mesenchymal cells may be regulated by a unique pattern of antigen nonspecific cell-cell interaction molecules. PMID- 3049624 TI - Sequence of contactin, a 130-kD glycoprotein concentrated in areas of interneuronal contact, defines a new member of the immunoglobulin supergene family in the nervous system. AB - The primary amino acid sequence of contactin, a neuronal cell surface glycoprotein of 130 kD that is isolated in association with components of the cytoskeleton (Ranscht, B., D. J. Moss, and C. Thomas. 1984. J. Cell Biol. 99:1803 1813), was deduced from the nucleotide sequence of cDNA clones and is reported here. The cDNA sequence contains an open reading frame for a 1,071-amino acid transmembrane protein with 962 extracellular and 89 cytoplasmic amino acids. In its extracellular portion, the polypeptide features six type 1 and two type 2 repeats. The six amino-terminal type 1 repeats (I-VI) each consist of 81-99 amino acids and contain two cysteine residues that are in the right context to form globular domains as described for molecules with immunoglobulin structure. Within the proposed globular region, contactin shares 31% identical amino acids with the neural cell adhesion molecule NCAM. The two type 2 repeats (I-II) are each composed of 100 amino acids and lack cysteine residues. They are 20-31% identical to fibronectin type III repeats. Both the structural similarity of contactin to molecules of the immunoglobulin supergene family, in particular the amino acid sequence resemblance to NCAM, and its relationship to fibronectin indicate that contactin could be involved in some aspect of cellular adhesion. This suggestion is further strengthened by its localization in neuropil containing axon fascicles and synapses. PMID- 3049631 TI - Fractures and dislocations of the distal radioulnar joint. AB - Fractures and dislocations of the distal radioulnar joint are frequently visualized as a secondary problem in comparison to the more apparent radius fractures. Frequently, in the long-term follow-up of patients with radius fractures, ulnar wrist pain secondary to distal radioulnar joint incongruity is the final outcome. Therefore, in the evaluation of the injured forearm the distal radioulnar joint must be assessed clinically and radiographically. In this assessment if distal radioulnar joint instability or incongruity is present then joint stabilization or reduction, respectively, must be attained. PMID- 3049630 TI - Classification and management of intra-articular fractures of the distal radius. AB - A classification of distal radial articular fractures is described, based on observations of consistent patterns of fracture fragmentation and displacement. The classification categorizes articular fractures into four types, with the medial complex assuming a pivotal position as the cornerstone of both the radiocarpal and distal radioulnar joints. The purpose of this classification is four-fold: (1) to afford identification and an understanding of the displacement characteristics of the major fracture components, (2) to provide practical and rational guidelines for the management of these injuries based on specific fracture patterns, (3) to emphasize the frequency of concurrent soft tissue and other skeletal injuries associated with the more severe types of articular disruption, and (4) to serve as a prognostic gauge for the varied spectrum of distal radius articular injury. Optimal management of distal radius fractures necessitates the differentiation of articular from extra-articular fractures as well as prompt detection of unstable injuries. While the majority of unstable fractures can be successfully managed by closed methods, a substantial and increasing number require open treatment for restoration of articular congruity as well as repair of concomitant soft tissue and skeletal injuries. In all cases, precise reduction of the key medial fragments is essential to maximum recovery. PMID- 3049629 TI - Growth factor production by a human megakaryocytic tumor cell line. AB - A recently described human megakaryocytic tumor cell line was analyzed for the presence of growth factor activity and was found to produce large quantities of transforming growth factor beta-like (TGF-beta) and basic fibroblast growth factor-like (bFGF) activities. Growth factor activities were identified using a radioreceptor assay for the TGF-beta-like activity, a heparin-binding assay for the b-FGF-like activity, and a demonstration of distinct biological activities for each type of factor. Tumor poly-A+ RNA revealed strong signals when probed with complementary DNA corresponding to bovine basic FGF and human TGF-beta and weak signals when probed with cDNA corresponding to epidermal growth factor (EGF) and TGF-alpha. The levels of EGF and TGF-alpha produced in the tumor line were too low to be detected by radioreceptor assays. Relative levels of messenger RNA encoding each of the growth factors reflected the relative levels of each of the respective factors tested. These data represent the first definitive identification of FGF-like activities in megakaryocytic-like cell lines. Interestingly, the line displayed little activity similar to platelet-derived growth factor (PDGF) when assayed either biochemically or by poly-A+ RNA analysis. PMID- 3049632 TI - Reconstruction of post-traumatic deformity of the distal radius and ulna. AB - Proper patient selection, meticulous preoperative planning, and precise surgical technique are all necessary for a successful outcome following osteotomy of the distal radius. The patients should be young, manually active, and motivated. Radiocarpal post-traumatic arthritis or dystrophy are contraindications to this procedure. PMID- 3049633 TI - Treatment of comminuted distal radius fractures with pins and plaster. AB - Based on this review, it is evident that the anatomic results obtained by the use of pins and plaster for the treatment of comminuted distal radial fractures is insufficient to yield a high percentage of satisfactory functional end results. In addition, the high complication and reoperation rate noted in our series makes us question whether the technique of pins and plaster should remain a treatment option for these difficult fractures. Perhaps a combination of internal fixation with bone grafting combined with external fixation will provide a solution to this fracture, particularly in young, active patients. PMID- 3049634 TI - Barton's and Smith's fractures. AB - Smith's and Barton's fractures are discussed in light of their original descriptions and current experience. Included here are excerpts from the original descriptions of Smith and Barton as well as the management of these two kinds of fracture. PMID- 3049635 TI - Pitfalls and complications of fractures of the distal radius and ulna in childhood. AB - Fortunately, most bone forearm fractures in children heal with minimal functional disability. My approach to management of the most common injuries is presented. By following the guidelines outlined in this article, the generalist as well as the most sophisticated upper extremity surgeon should be able to avoid pitfalls and complications when managing forearm fractures in children. PMID- 3049636 TI - Injury to the immature carpus. AB - Traumatic injury to the immature carpus is infrequent because of the sequence of carpal ossification, the resiliency of surrounding ligaments, and the vulnerability of the radius to fracture. Diagnosis requires careful palpation and suspicion, particularly in younger children. Treatment should consider the healing, growth, and modeling potential of the carpus. PMID- 3049637 TI - Stabilization of fractures in the hand and wrist with traumatic soft tissue and bone loss. AB - Open type III fractures of the hand or wrist with severe bone and soft tissue loss justify aggressive treatment to restore anatomy, assure healing, and maximize functional recovery. The techniques of modern wound excision used at initial surgery predictably result in a decompressed and surgically clean wound within a few days from injury in the vast majority of cases. This allows a safe application of delayed primary internal fixation and bone grafting for fracture restoration or joint arthrodesis as well as early wound closure or coverage. The immediate or early application of stable external devices, internal fixation, or combinations of the two along with early bone grafting restores the structural integrity of the skeleton, reduces pain, protects other repaired and reconstructed tissues, promotes the healing, and supports early and intensive functional rehabilitation of the hand and wrist. Early wound closure or coverage minimizes scar formation. Together, the early sequencing of effective wound debridement with skeletal stabilization and bone grafting and early wound closure or coverage provide the most favorable circumstances for healing and functional recovery of the seriously damaged hand and wrist. PMID- 3049638 TI - Scaphoid nonunion. AB - Scaphoid nonunion is common, but the exact pathophysiology of this complication is unclear. Explanations include lack of treatment, poor initial treatment, delay in diagnosis, synovial fluid dynamics, precarious vascularity, fracture displacement, and carpal instability. Currently, the diagnosis is best confirmed by classic changes on plain radiographs, instability testing, arthrography, and arthroscopy in selected cases. Nine carpal bones are not benign. The natural history of scaphoid nonunion is one of progressive arthritis. Attempts at obtaining bony healing are therefore recommended. In treating established nonunions without arthritis, the Russe bone graft technique is the mainstay of treatment. A union rate of 90 per cent is to be expected. Electrical stimulation is an alternative when there is no synovial pseudarthrosis or scaphoid collapse deformity, or if a previous Russe graft has failed. If a significant humpback scaphoid or collapse deformity is present, internal fixation with the Herbert screw and scaphoid reconstruction with a bone graft are our choices. Healing rates are less than those with the Russe graft, but one may achieve improved motion of the wrist and earlier return to function. When scaphoid nonunion is accompanied by degenerative arthritis, salvage procedures are recommended. Radial styloidectomy is a simple procedure that will preserve motion and buy time. Soft tissue interposition with excision of a small proximal pole is useful, particularly if no collapse deformity is present. Silicone replacement alone has fallen into increasing disfavor because of the high incidence of subluxation and silicone synovitis. Combining silicone replacement with intercarpal fusion (the SLAC procedure) may lessen these complications. Proximal row carpectomy is another procedure that may preserve motion, though often at the expense of weakness, particularly in the younger patient requiring significant grip strength. In these cases, standard wrist fusion seems the most predictable alternative. PMID- 3049639 TI - Management of greater arc carpal fractures. AB - Greater arc injuries are fracture dislocations that involve the perilunar carpal bones. The commonest of these injuries is the dorsal transscaphoid perilunate fracture dislocation. The recommended treatment for acute injuries is open reduction and Kirschner wire fixation of the scaphoid fracture through a dorsal midline approach. Stability to the midcarpal joint is thus provided and no further pins are needed. Established scaphoid nonunions are treated by bone grafting and Herbert screw fixation through a volar approach. Scaphocapitate fracture syndrome is treated by open reduction and pin fixation of the displaced capitate fragment through the dorsal approach. If the scaphoid is displaced it is also openly reduced and pinned. PMID- 3049640 TI - Recognition and treatment of uncommon carpal fractures. AB - Fractures of the trapezial ridge, hamate hook, and pisiform are often due to isolated, direct injury, but other carpal bone fractures are seldom isolated injuries. Midcarpal fractures should stimulate a search for associated perilunate injury; distal carpal row fractures suggest carpometacarpal fracture- dislocations or subluxations. Crush injury with fracture of the hamate, triquetrum, or trapezium may indicate an axial subluxation of the ulnar or radial carpus. Bone scans can be useful as a screening tool. Diagnosis will often require special radiographic techniques such as tomography. Carpal bone fractures are usually intra-articular, and treatment should be aimed at restoring joint congruity. Small extra-articular fractures and injuries to the pisiform can be successfully treated with excision. PMID- 3049641 TI - Infected fractures of the hand and wrist. AB - Infected fractures of the hand and wrist are uncommon, occurring most often after open crushing injury in a contaminated environment. Fundamental principles of treatment include thorough debridement of necrotic material, appropriate antibiotic selection, and adequate stabilization of bone. Delayed reconstruction of bone and soft tissue is at times aided by the use of free tissue transfer. PMID- 3049642 TI - Intra-articular fractures of the basilar joint of the thumb. AB - This article on intra-articular fractures of the basilar joint of the thumb discusses the biomechanical and treatment complexities of fractures involving the thumb carpometacarpal joint. This review focuses on the treatment of thumb carpometacarpal fractures and dislocations, consolidating current philosophy and rationale regarding operative and nonoperative treatment. Treatment recommendations are based upon principles established in previous clinical and biomechanical studies emphasizing fracture-specific modalities. Emphasis is placed on maintenance of articular congruity, fracture stability, early motion, and maximum return of function. PMID- 3049643 TI - Management of malunited fractures of the metacarpal and phalangeal shafts. AB - Malunions of the tubular bones of the hand should be carefully studied to understand the original deforming forces at the time of injury. Once a three dimensional concept of the deformity is embraced, a careful plan for osteotomy can be developed and executed. Surgical approach must afford adequate access with the least possible injury to soft tissues. Technique of osteotomy must be tailored to the configuration, location, and biomechanical requirements for proper realignment of the malunited fracture. Important principles in the management of metacarpal and phalangeal malunions are: 1. Rotatory deformities are most disabling yet frequently not appreciated. A 10-degree rotational malalignment in the metacarpal results in a 2-cm overlap at the finger tip. Alignment should always be checked with the fingers flexed into the palm. 2. Adherence to biomechanical principles of fracture repair is mandatory. 3. The appropriate form of osteotomy and subsequent fixation must be carefully chosen and applied to each individual deformity. Familiarity with the osteotomy techniques and alternative forms of fixation affords flexibility in managing complex deformities. 4. The presence of malunion suggests the presence of fracture disease in the soft tissues. They must be delicately handled and carefully inspected for the presence of scarring, adhesions, and contractures. Careful protection of delicate structures should be complemented by judicious tenolysis and arthrolysis at the time of osteotomy. 5. Appropriate, functional postoperative rehabilitation is mandatory. Without adequate therapy the best surgery will produce suboptimal results. Adherence to these principles can avoid further complications and problems and provide a successful ultimate outcome. PMID- 3049644 TI - Fractures of the distal phalanx. AB - Fractures of the distal phalanx, except for those of the articular surface, are sustained in crushing injuries and as such require care for the surrounding soft tissues and rarely need specific treatment for the fracture itself. Displaced articular fractures on the palmar side, however, are associated with avulsion of the flexor digitorum profundus tendon and will need careful replacement by surgical means. While there is some disagreement among authorities, it is believed here that the dorsal articular fracture, the mallet fracture, can and should be treated by nonoperative means. PMID- 3049646 TI - Preparative chromatography of proteins analysis of the multivalent ion-exchange formalism. AB - Multivalent ion exchange is proposed as a generally applicable formalism to describe the non-linear chromatographic adsorption of biopolymers. Single- and multi-component isotherms are calculated that explicitly account for the influence of the mobile phase modulator concentration. Three regions of binding (strong, intermediate, and weak) are distinguished on the basis of strength of interaction, and the potential advantages of operating in the strong region for preparative purposes are pointed out. The inapplicability of the Langmuir isotherm in this region is demonstrated, and separation schemes that take advantage of the characteristic features of biopolymeric adsorption are described. The rectangular single-component ion-exchange isotherms are shown to reduce in the presence of competition to reversible concave-down forms. PMID- 3049645 TI - Simultaneous determination of the prodrug zofenopril and its active drug in plasma by capillary gas chromatography-mass-selective detection. AB - After oral administration of zofenopril, the active sulfhydryl angiotensin converting enzyme inhibitor is released. Zofenopril is currently under clinical investigation as an antihypertensive. Blood samples are reacted with N ethylmaleimide, immediately after collection, processed into plasma and stored frozen for subsequent analysis. After addition of two internal reference standards, one each for the prodrug and the active compound, the plasma samples are purified by a combination of liquid-liquid and solid-phase extractions. The dried methylated extracts are reconstituted with tetramethylbenzene and chromatographed by automated splitless injection on a fused-silica capillary column, connected to a mass-selective detector. The analytes and the internal reference standards are chromatographically resolved and a common fragment ion is monitored for the analytes. A limit of quantitation of approximately 1 ng/ml of plasma is achieved. PMID- 3049647 TI - Surface-mediated retention effects of subtilisin site-specific variants in cation exchange chromatography. AB - Wild-type subtilisin and several site-specific variants were resolved on a strong cation-exchange column by isocratic elution and a series of sodium chloride concentrations. Changes in primary sequence at the protein surface have an observable effect on the chromatographic behavior of subtilisin. This supports the concept that three-dimensional structure determines which biopolymer surface residues are in position to interact with the stationary phase surface. The retention data fit the stoichiometric displacement model (SDM) of retention. Plots of ln k' vs. ln 1/[NaCl] yield values for the average number of ionic groups (Z) on the protein that interact with the support matrix. Application of the SDM to the chromatographic retention of the variants has uncovered an unusual phenomenon at the protein surface at low ionic strength. A SDM plot normally provides a linear relationship between ln k' and ln 1/[NaCl] with the slope corresponding to the Z number. This study revealed two lines differing in slope and intercept, indicating that the Z number of subtilisin changes at some intermediate ionic strength of the eluent. These results are attributed to some salt-induced protein surface event that triggers a change in structure. Chromatographic detection of this occurrence reflects the connection between the surface-mediated event and mobile phase ionic strength. PMID- 3049648 TI - Structural characterization of glycoproteins. AB - The structural characterization of glycoproteins by high-performance liquid chromatography (HPLC) is often difficult because of microheterogeneity of their oligosaccharide groups. To investigate this phenomenon, a series of human plasma glycoproteins of known amino acid sequence and carbohydrate structure was subjected to comparative study by HPLC. Methods for the isolation of singly glycosylated glycopeptides were developed. The chromatographic behavior in reversed-phase HPLC of mono-, di-, and multiglycosylated glycopeptides was compared with that of unglycosylated peptides of known amino acid sequence. Glycopeptides tended to be eluted earlier than non-glycopeptides, but the major factor governing retention was hydrophobicity. As shown by comparative study of five plasma glycoproteins by four chromatographic methods, microheterogeneity of the oligosaccharides affects chromatographic characterization of glycoproteins. In anion-exchange HPLC, carbohydrate charge heterogeneity is reflected by the broadening or asymmetry of peaks. Hydroxyapatite chromatography is useful for the purification of several forms of ceruloplasmin and other glycoproteins. The effect of carbohydrate is small in reversed-phase chromatography, but some proteins are denatured by the stringent conditions. Carbohydrate does not have much effect in hydrophobic interaction chromatography, which is more gentle. Because the chromatographic behavior of a glycoprotein may vary significantly with the procedure applied, several types of HPLC methods should be used for the characterization of glycoproteins. PMID- 3049649 TI - Application of high-performance liquid chromatography in neurophysin disulfide assignment. AB - The combined use of ion-exchange, and reversed-phase high-performance liquid chromatography (HPLC) for the isolation of cystine-containing peptides from highly heterogeneous products of the proteolytic digestion of bovine neurophysins is described. The protein was sequentially cleaved by enzymes of decreasing specificity; the peptides released were initially fractionated by gel chromatography and then purified by HPLC. The purified peptides were analyzed by determination of their amino acid composition and mass spectrometry, supported by sequencing techniques. Three of the seven disulfide pairs of neurophysin have now been assigned. The usefulness of the combined use of HPLC and mass spectrometry in assigning these and the other disulfide pairs is illustrated. PMID- 3049650 TI - Separation of proteins by reversed-phase high-performance liquid chromatography. II. Optimizing sample pretreatment and mobile phase conditions. AB - The effects of separation variables such as temperature, pH and composition of the mobile phase (including additives such as chaotropes, ion-pairing agents and surfactants), sample size and sample pretreatment for reversed-phase high performance liquid chromatography (RP-HPLC) of proteins is examined. Experimental optimization of these parameters using the preferred instrumental and column conditions described previously lead to well behaved chromatographic performance for most proteins. This allowed us to achieve the required level of performance for the first dimension (RP-HPLC) separation of most protein samples by the chromatophoresis process. PMID- 3049651 TI - Multicenter evaluation of four methods of yeast inoculum preparation. AB - We initiated a comparative study of four methods of yeast inoculum preparation: a spectrophotometric method, the Wickerham card method, a hemacytometer method, and the Prompt inoculation system. The variability in inoculum size obtained when each method was applied to two strains each of Candida albicans, Candida tropicalis, Candida parapsilosis, Torulopsis glabrata, Cryptococcus neoformans, and Saccharomyces cerevisiae was analyzed in a single laboratory. Each method was performed in triplicate on the same day and on three separate days to provide estimates of within-day and between-day variations. Inoculum size was determined by viable colony counts. The greatest range of inoculum sizes was seen with the Wickerham card method. Viable counts ranged from 1.1 X 10(6) to 24.2 X 10(6) CFU/ml among the 12 yeast isolates. The greatest variation was observed with the Prompt system. Within-day coefficients of variation averaged 19% (range, 4 to 45%), and between-day coefficients of variation averaged 22% (range, 3 to 51%). Variation between laboratories was evaluated by comparing inoculum values obtained by each method in three different laboratories for two strains of C. albicans. The spectrophotometric method was the least variable and the Wickerham card and hemacytometer methods were the most variable methods between laboratories. The spectrophotometric method is recommended as the method of choice for preparation of a standardized inoculum suspension for susceptibility testing of yeasts. PMID- 3049652 TI - False-positive DNA probe test for Legionella species associated with a cluster of respiratory illnesses. AB - Between 11 November 1986 and 28 February 1987, legionellosis was diagnosed in 23 patients at one hospital with a recently marketed Legionella-specific DNA probe for respiratory secretions. Only 10 of the 23 probe-positive patients showed findings typical of Legionella pneumonia, including a temperature of greater than or equal to 100.5 degrees F (approximately 38.1 degrees C) and radiographic evidence of pneumonia. No differences were found in the results of laboratory studies, demographic features, or underlying risk factors for these 10 probe positive patients when compared with the 13 probe-positive patients with nonpneumonic illnesses. A case-control study comparing probe-positive and negative patients failed to identify any different features of disease or epidemiologic characteristics. Probes of repeat specimens of sputum were still positive 2 to 13 weeks after the initial test in 5 (50%) of the 10 probe-positive patients. The clinical features in most patients were atypical for legionellosis, and the diagnosis could not be confirmed by traditional laboratory tests performed on duplicate specimens processed at the Centers for Disease Control. This report emphasizes the need for clinical microbiology laboratories to confirm test results from new procedures by accepted diagnostic methods. PMID- 3049653 TI - Multiple Candida strains in the course of a single systemic infection. AB - Species and strain variabilities have been monitored during the history of a prolonged Candida infection in a single compromised bone marrow transplant patient by analyzing sugar assimilation patterns, high-frequency switching repertoires, and Southern blot hybridization patterns with two cloned mid-repeat sequences (Ca3 and Ca7) which are species specific for Candida albicans and one cloned mid-repeat sequence (Ct13-8) which is species specific for Candida tropicalis. Evidence is presented that during the course of this infection (i) two strains of C. albicans and three strains of C. tropicalis were distinguished by their switching repertoires, Southern blot hybridization patterns, and sugar assimilation patterns; (ii) the three C. tropicalis strains were in a high frequency mode of switching; (iii) two C. tropicalis strains coexisted in the blood and three C. tropicalis strains coexisted in the throat at different times during the history of the infection; (iv) amphotericin B treatment selectively removed one of two C. tropicalis strains coexisting in the blood and this strain exhibited greater susceptibility to amphotericin B in vitro (the remaining strain was subsequently removed from the blood by flucytosine treatment); and (v) both the strain removed from the blood by amphotericin B and the strain removed from the blood by flucytosine reappeared several days later at another site of infection. It is demonstrated for the first time that C. tropicalis is capable of high-frequency switching of colony morphology just as C. albicans is, that there is more than one strain-specific switching repertoire in C. tropicalis, and that a C. tropicalis mid-repeat sequence can be used for discriminating species and assessing strain relatedness, as previously demonstrated for C. albicans mid repeat sequences. PMID- 3049654 TI - Virulence of Escherichia coli in relation to host factors in women with symptomatic urinary tract infection. AB - The relationship between bacterial characteristics and the severity of urinary tract infection in adults has not been clarified. In this study, Escherichia coli strains (n = 178) were prospectively collected from women with community-acquired urinary tract infection. The isolates were identified by O:K:H serotype and characterized for adherence, hemolysin production, and serum bactericidal resistance. The patients had acute pyelonephritis with or without complicating factors and acute cystitis. Nine serotypes (O1:K1:H7, O1:K1:H-, O2:K1:H-, O4:K12:H1, O7:K1:H-, O9:K34:H-, O16:K1:H6, O16:K1:H-, and O75:K5:H-) comprised 65% of the strains in uncomplicated pyelonephritis, but were significantly less often encountered in complicated pyelonephritis or cystitis. Adherence was the single property most characteristic of the pyelonephritogenic clones. Adhesins specifically recognizing Gal alpha 1----4Gal beta-containing receptors occurred in 80% of strains in uncomplicated pyelonephritis, in 50% of strains in complicated infections, and in 37% of cystitis strains. Hemolysin production and serum resistance did not correlate with any disease pattern. Advanced age did not seem to reduce the selection of virulent E. coli to cause pyelonephritis. These results demonstrate in women a relationship between E. coli virulence and the severity of urinary tract infection analogous to that previously observed in pediatric populations and also illustrate the balance between host resistance and bacterial virulence in the urinary tract. PMID- 3049656 TI - Comparison of immunofluorescence, enzyme immunoassay, and Western blot (immunoblot) methods for detection of antibody to human T-cell leukemia virus type I. AB - A total of 3,349 serum samples were screened by the immunofluorescence (IF) method for antibody to human T-cell leukemia virus type I (HTLV-I). Only 9 of 2,409 specimens from selected individuals, blood bank donors, patients with encephalitis-meningitis, and human immunodeficiency virus antibody-positive homosexual or bisexual men were reactive by IF. In addition, 940 serum samples from intravenous drug abusers were tested by IF and also by an HTLV-I enzyme immunoassay (EIA) method. Of these, 222 (24%) were positive for both HTLV-I and HTLV-II antigens by IF, and 191 of these 222 were also reactive in the HTLV-I EIA. Of the 31 IF-positive, EIA-negative serum samples, 20 exhibited optical density readings greater than or equal to 70% of the positive cutoff in the EIA, and 29 samples reacted with 1 or more bands in the Western blot (immunoblot) test. An additional 10 specimens that were EIA negative reacted only with HTLV-I by IF. Differences in staining morphology and in reactions on HTLV-I and HTLV-II antigens before and after absorption of the serum specimens with HTLV-I and HTLV II-infected cell pellets revealed six distinct serological patterns by IF. These results indicate that infections by HTLV-I or by another closely related retrovirus(es) occur in California. Further studies utilizing statistically valid sampling methods are needed to estimate true prevalence rates among various groups. IF and Western blot tests should supplement the EIA method to maximize sensitivity and specificity of test procedures. PMID- 3049655 TI - Clonal origin, restricted natural distribution, and conservation of virulence factors in isolates of enterotoxigenic Escherichia coli serogroup O126. AB - Enterotoxigenic Escherichia coli serogroup O126 isolates have been isolated in Hong Kong since 1982 from sporadic cases of infantile diarrhea and from one outbreak in a neonatal ward. A 64-megadalton plasmid encoding colonization factor antigen I and heat-stable enterotoxin was identified in all 23 isolates. Enterotoxigenic E. coli strains producing heat-stable enterotoxin from different regions of Southeast Asia were collected and compared by biotyping, antibiotic resistance patterns, and plasmid profiles. Restriction endonuclease digestion of plasmids and subsequent Southern blot analysis with the heat-stable enterotoxin gene probe of representative strains showed a unique plasmid was harbored by all heat-stable enterotoxin-producing O126 strains tested. These results are consistent with conservative inheritance of enterotoxin plasmids within enterotoxigenic E. coli strains over a 2-year period in Hong Kong. PMID- 3049657 TI - Novel application of video image processing to biochemical and antimicrobial susceptibility testing. AB - ALADIN (Analytab Products, Plainview, N.Y.) is an automated instrument that uses video imaging (computer-assisted guided video camera) for the determination of biochemical and antimicrobial susceptibility test reactions. This collaborative investigation compared video-generated results obtained with ALADIN with visually determined findings. Both approaches were used to view identical reactions. Overall agreement for biochemical and antimicrobial susceptibility tests was greater than 95%. This study demonstrates that video imaging is an acceptable approach for determining microbial responses to biochemical and antimicrobial agents and may provide, with appropriate computer modifications, more accurate and reproducible results than are possible by visual scrutiny. PMID- 3049658 TI - Occurrence of mucoid M-18 Streptococcus pyogenes in a central Ohio pediatric population. AB - During a 1-year period from October 1986 through September 1987, we recovered 116 mucoid, hemolytic Streptococcus pyogenes isolates from clinical specimens collected from patients seen at our pediatric institution. A total of 102 isolates were from throat cultures (101 for pharyngitis, 1 for acute rheumatic fever), 13 were from other superficial body sites, and 1 was from pleural fluid. All of 40 mucoid isolates tested to date were determined to be M-type 18 strains. A direct latex agglutination test for group A carbohydrate antigen in throat swab specimens was equally sensitive in detecting M-18 mucoid and nonmucoid strains (45 of 77 [58%] and 795 of 1,186 [67%], respectively; not significant, P greater than 0.05). Antimicrobial susceptibility tests performed with 40 mucoid and 40 nonmucoid isolates against penicillin and nine other antimicrobial agents showed all strains to be susceptible, with no difference in MICs. All isolates tested were also considered fully susceptible to the bactericidal activity of penicillin. Further studies are needed to establish the relative virulence of M 18 strains and their possible association with the resurgence of acute rheumatic fever in central Ohio and other areas of the United States. PMID- 3049661 TI - Evaluation of a new latex test and a new enzyme immunoassay for determination of rubella immunity. AB - A new enzyme immunoassay (Rubenostika; Organon Teknika, Turnhout, Belgium), a new latex agglutination test (Rubalex; Orion Diagnostica, Espoo, Finland), and three other accepted methods for the determination of rubella immunity were compared with a standard hemagglutination inhibition assay. Of 224 serum samples tested, 54 (24%) were nonreactive and 24 (11%) were low titered. All procedures were very specific (94 to 100%). Rubenostika was the least sensitive method (88%), and Rubalex was the most sensitive (98%). PMID- 3049660 TI - Human-isotype-specific enzyme immunoassay for antibodies to pneumococcal polysaccharides. AB - A simple enzyme immunoassay has been developed to allow the quantitation of the human response to immunization with pneumococcal polysaccharide. The assay uses the 14-valent vaccine (Pneumovax) as a convenient antigen to adsorb to the solid phase microdilution plate wells and commercially available isotype-specific antibody conjugates. The results have been expressed as arbitrary pneumococcal polysaccharide antibody units by reading off a standard curve constructed by using heterogeneous pooled serum. All nonimmunized subjects tested had immunoglobulin G (IgG) antibodies present in serum. All six control subjects who were immunized with Pneumovax demonstrated an IgG response, and the majority responded with a rise in IgA- and IgM-specific antibody concentrations at a mean of 6 weeks postimmunization. Five out of six cord sera tested contained IgG antibodies only, which were present in concentrations similar to those seen in adults, whereas in 6- to 12-month-old infants only low levels of IgG and IgM and no IgA antibodies were detected. Serum taken from 10 hypogammaglobulinemic patients immediately prior to infusion of immunoglobulin showed low to negative IgG antibody concentrations, and no IgA or IgM antibody was present. PMID- 3049663 TI - Cellular fatty acid composition of Kingella species, Cardiobacterium hominis, and Eikenella corrodens. AB - We determined the cellular fatty acid composition of reference strains and clinical isolates of each of the three Kingella species, Cardiobacterium hominis, and Eikenella corrodens by using capillary gas chromatography. Kingella denitrificans and Kingella kingae contained myristic (14:0) and palmitic (16:0) acids as major acids, whereas cis-vaccenic (18:1 omega 7c) and palmitic acids were the major acids in Kingella indologenes, C. hominis, and E. corrodens. C. hominis differed from the other four species by the absence of 3-hydroxylauric (3 OH-12:0) acid, from K. indologenes by the presence of 3-hydroxypalmitic (3-OH 16:0) acid, and from E. corrodens by the presence of 3-hydroxymyristic (3-OH 14:0) acid. E. corrodens contained a small amount (2%) of myristic acid, while the other four species contained moderate to large amounts (11 to 31%) of this acid. PMID- 3049662 TI - Evaluation of autoscan-4 for identification of members of the family Enterobacteriaceae. AB - A study was performed to compare the Autoscan-4 (MicroScan, Inc., Mahwah, N.J.) with conventional biochemical methods for identifying clinical isolates of the family Enterobacteriaceae. The Autoscan-4 yielded correct identification of 95.4% of the isolates at the species level and 98.4% at the genus level. Only one misidentification was observed. The identification of both common and less-common isolates of Enterobacteriaceae makes this system highly efficient. PMID- 3049659 TI - Enzyme-linked immunosorbent assay for Escherichia coli heat-stable enterotoxin type II. AB - The gene for Escherichia coli heat-stable enterotoxin type II (STII) was fused to the genes for protein A from Staphylococcus aureus and beta-galactosidase in two different expression systems. Antibodies raised in rabbits against the protein A STII fusion protein recognized the beta-galactosidase-STII fusion protein. The latter fusion protein was used as the immobilized antigen in an enzyme-linked immunosorbent assay (ELISA) for detection of STII. The correlation between the results of the ELISA and the intestinal loop test in piglets was 95%, suggesting that the ELISA can be used to reliably detect STII. PMID- 3049665 TI - Chronic relapsing experimental allergic encephalomyelitis in the Lewis rat: studies on immune regulation. AB - To study immunoregulation of chronic relapsing experimental allergic encephalomyelitis (CR-EAE) in Lewis rats, we adoptively transferred concanavalin A-activated lymph node cells (LNC) or splenocytes, from hind footpad-inoculated donors at the onset (day 11), or recovery (day 16), of the first attack. Popliteal LNC, especially from day 16 donors, provided significant and dose dependent, but incomplete, protection of recipients from encephalitogenic challenge; maximal mean delay in EAE onset was 10 days later than controls, with subsequent paralysis reduced more than 6-fold. In contrast, particularly from day 11 donors, superficial inguinal LNC recipients developed actively induced disease of normal severity up to 4 days earlier than CR-EAE controls. Furthermore day 11 EAE splenocytes, but not day 16 ones, adoptively transferred disease into 50-88% of naive recipients. In separate studies, we demonstrated unresponsiveness to active induction of disease in all rats re-challenged during stable late remission, as well as in a minority of animals pretreated with antigen in incomplete Freund's adjuvant. These results suggest an organ-dependent and time dependent balance between effector and suppressor populations in the model. PMID- 3049666 TI - Fluorescent voltage-sensitive dyes: applications for neurophysiology. AB - Voltage-sensitive dyes are a means to optically monitor changes in membrane potential. Their application in research has grown steadily over the last two decades as better dyes have been developed. The techniques presently in use are providing unique information about biologic systems from bacteria to the functional organization of primate occipital cortex. This review provides a history of the dyes, the data supporting their voltage sensitivity, and the techniques required for their use. The limitations in using and interpreting the voltage-sensitive dyes, as well as their diverse applications in all areas of research, especially neurophysiology, are comprehensively presented. PMID- 3049664 TI - DNA probes to identify Shiga-like toxin I- and II-producing enteric bacterial pathogens isolated from patients with diarrhea in Thailand. AB - When Shigella species, Escherichia coli, and five other bacterial enteric pathogens isolated from children with diarrhea in Thailand were tested for hybridization under stringent conditions with probes for Shiga-like toxins I and II, only 30 Shigella dysenteriae 1 hybridized with the Shiga-like toxin I probe. PMID- 3049667 TI - Surgical monitoring with auditory evoked potentials. AB - This comprehensive review of surgical monitoring with auditory evoked potentials (AEPs) includes a detailed discussion of techniques used for recording brainstem auditory evoked potentials, direct eight-nerve potentials, and electrocochleograms. The normal waveform of these different potentials is discussed, and the typical patterns of abnormalities seen with different insults to the peripheral or central auditory pathways are presented. The mechanisms most probably responsible for changes in AEPs during surgical procedures are analyzed. A critical analysis is made of what represents a significant change in AEPs. Also considered is the predictive value of intrasurgical changes of AEPs. Finally, attempts are made to determine whether AEPs monitoring can assist the surgeon in the prevention of postsurgical complications. PMID- 3049668 TI - Medial femoral torsion and osteoarthritis. AB - We investigated the hypothesis that medial femoral torsion is a predisposing factor to osteoarthritis of the hip. Anteversion was measured by biplane radiography in 44 hips (32 patients) with idiopathic osteoarthritis of the hip and a control group of 98 normal hips (49 adults). The mean and range of anteversion measures were similar in both groups. The mean was 22 degrees for osteoarthritic hips (range 3 degrees-49 degrees) and 19 degrees for normal controls (range -2 degrees-49 degrees). This difference was not significant. This study disagrees with others that propose an association between increased anterversion and osteoarthritis of the hip. Prophylactic operative correction of femoral torsion for osteoarthritis is not recommended. PMID- 3049669 TI - Role of radionuclide imaging in the diagnosis of acute osteomyelitis. AB - Over the last decade, the role of nuclear medicine studies in the diagnosis of acute osteomyelitis has been discussed in depth in the literature. Yet, the respective roles played in this setting by each of the commonly used radionuclide studies often are confusing. In an attempt to develop a cogent diagnostic strategy, we reviewed the literature published within the last 12 years pertaining to the use of radiophosphate bone scintigraphy as well as gallium and indium WBC imaging in the diagnosis of this condition. Based on our findings, we propose an alternative approach to the evaluation of a patient with suspected acute osteomyelitis. PMID- 3049670 TI - Monostotic fibrous dysplasia of the lumbar spine: case report and review of the literature. AB - Fibrous dysplasia has been frequently reported to involve the spine in the polyostotic form, but only rarely has monostotic fibrous dysplasia been noted. In the only previously reported case involving the lumbar spine, the disease was confined to the transverse process. The present case demonstrates monostotic fibrous dysplasia involving the vertebral body in addition to the posterior elements. The plain radiographic, computerized tomography, and histologic examinations are presented. PMID- 3049673 TI - Cultured human endothelial cells stimulated with cytokines or endotoxin produce an inhibitor of leukocyte adhesion. AB - Activation of cultured human endothelial cells (HEC) by inflammatory stimuli, such as interleukin 1 (IL-1), tumor necrosis factor (TNF), and bacterial endotoxin (lipopolysaccharide, LPS), increases their surface adhesiveness for blood leukocytes and related cell lines. We now report that activated HEC also generate a soluble leukocyte adhesion inhibitor (LAI), which accumulates in conditioned media from IL-1-, TNF-, or LPS-treated, but not sham-treated, HEC cultures. LAI significantly inhibits the adhesion of PMN and monocytes to activated, but not unactivated, HEC. In contrast, LAI has no effect on the adhesion of lymphocytes, the promyelocytic cell line HL-60 or the monocyte-like cell line U937 to HEC monolayers. LAI appears to act directly on the leukocyte, but does not inhibit either agonist-induced responses in PMN (membrane depolarization, changes in cytosolic calcium concentration, superoxide production) or PMN attachment to serum-coated plastic surfaces. Endothelial generation of LAI is blocked by actinomycin D but not by aspirin or indomethacin. Preliminary biochemical characterization indicates that LAI is a soluble, protein containing molecule that is heat- and acid-stable. Fractionation by HPLC gel filtration yields a single peak of LAI activity (14,000 less than Mr greater than 24,000). Thus, in addition to proadhesive cell surface changes, the endothelium may also actively contribute to the regulation of endothelial-leukocyte interactions at sites of inflammation in vivo through the production of soluble adhesion inhibitors such as LAI. PMID- 3049672 TI - Studies on the molecular mechanisms of human Fc receptor-mediated phagocytosis. Amplification of ingestion is dependent on the generation of reactive oxygen metabolites and is deficient in polymorphonuclear leukocytes from patients with chronic granulomatous disease. AB - Human PMN and monocytes both possess a mechanism for amplifying Fc receptor mediated phagocytic function, which is dependent on activation of the respiratory burst. The pathway for augmentation of phagocytosis requires superoxide anion, hydrogen peroxide, and lactoferrin and is independent of the hydrogen peroxide MPO-halide system. In neither cell type is this mechanism induced upon exposure to the opsonized target. PMN require an additional signal for stimulation of the respiratory burst; this is not true of monocytes. On the other hand, monocytes require an exogenous source of lactoferrin in order to activate this pathway for enhanced ingestion. The dependence of this pathway for both PMN and monocytes on superoxide anion, hydrogen peroxide, and cell-bound lactoferrin is consistent with a role for locally generated reactive oxygen metabolites, possibly hydroxyl radicals, in phagocytosis amplification. Patients with chronic granulomatous disease, who are genetically deficient in the ability to activate the respiratory burst, are unable to amplify Fc receptor-mediated phagocytosis. Thus, these patients may have a previously unrecognized defect in the recruitment of phagocytic function at inflammatory sites. PMID- 3049671 TI - The insulin receptor and the molecular mechanism of insulin action. PMID- 3049676 TI - Replacement of the damaged interarticular disc of the TMJ. AB - Meniscectomy should be abandoned and the removed, damaged disc of the TMJ should be replaced in patients suffering from internal derangement of the TMJ. In seventeen patients the disc which was removed, was replaced with fresh autogenous auricular cartilage. The follow-up period ranged from one to six years with a mean of almost three years. From the analysis of the results we concluded that all patients showed a postoperative improvement in their symptoms and joint function and that the use of intermaxillary fixation for a short period postoperatively had a significant effect on the subsequent increase in mouth opening. PMID- 3049674 TI - Human recombinant granulocyte-macrophage colony stimulating factor and interleukin 3 have overlapping but distinct hematopoietic activities. AB - The hematopoietic stimulatory activities of human recombinant IL-3 and granulocyte-macrophage colony stimulating factor (GM-CSF) were directly compared using highly enriched human bone marrow progenitor target cells. IL-3 supported a larger number of erythroid and megakaryocytic progenitor cells than did GM-CSF, while GM-CSF supported more myeloid progenitors. IL-3 directly stimulated the division and migration of primitive erythroid burst forming units, while GM-CSF merely sustained their net survival in culture without promoting division and expansion. IL-3 promoted the formation of larger numbers of multipotential granulocyte-erythroid-macrophage-megakaryocyte colony forming unit--derived colonies than did GM-CSF. These data indicate that human IL-3 and GM-CSF have overlapping but distinct hematopoietic activities, and suggest a potential role for the clinical application of combined IL-3/GM-CSF therapy. PMID- 3049677 TI - Tumours of the head and neck in children. A clinico-pathological analysis of 1,007 cases. AB - Tumours of the head and neck in children are uncommon, representing only 2-3% of all head and neck tumours. During the twenty year period 1964-1983, 12,876 childhood tumours were submitted for pathological diagnosis. Of these 1,007 (7.8%) were in the head and neck region, and it is this group that has been analysed. 30.6% (308 cases) were malignant neoplasms, 27.8% (280 cases) were benign neoplasms, 24.2% (244 cases) presented as tumour-like conditions and 17.4 (175 cases) were dysplasias arising from embryonal remnants. The overall sex ratio was 1.5:1 in favour of males. Lymphomas accounted for 15.9% overall (52.3% of the malignant neoplasms). Of benign tumours, haemangiomas were the most frequent (38.5%) and of the tumour-like conditions, dermoid and epidermoid cysts accounted for 36.1%. Of the embryonal remnant dysplasias, thyroglossal duct cysts accounted for 71.4%. PMID- 3049679 TI - Self-report measures for use with children: a review and comment. AB - This article underscores the need for self-report instruments for children to complement the teacher and parent questionnaires traditionally used to assess various aspects of children's psychological lives. Some of the problems inherent in using teachers, parents, and children as informants are delineated. Many self report instruments, in particular those that are used to assess children's self concept, anxiety, depression, and personality, are reviewed. The Children's Self Report Questionnaire (SRQ) was designed to assist in the diagnosis and detection of psychological deviance in 7- to 12-year-old children. The SRQ is easily administered, has broadly based norms, and has acceptable reliability and validity. The SRQ can be used as an aid to both research and clinical assessment and may provide insight into the inner world of the child. PMID- 3049678 TI - Chondrosarcoma of the mandible. Report of case and a survey of 23 cases in the Japanese literature. AB - A huge chondrosarcoma of the mandible (80 X 95 X 100 mm in size) with extension into the infratemporal fossa is described. The tumour was successfully treated by surgical removal and postoperative irradiation. A survey of the Japanese literature revealed 23 cases of chondrosarcoma with involvement of the mandible. The tumours occurred equally in males and females whose mean age was 38 years. The molar region was the site of predilection. The most common symptom was swelling and it was accompanied by pain in 7 cases and paraesthesia in 5 cases. Radiographically, the lesions were quite variable and with the exception of 3 cases in which information was not available, they consisted of a combination of irregular radiopacity and radiolucency in 9 cases, whereas the predominant feature was radiopacity in 6 cases and radiolucency in 4 cases. There was no radiographical abnormality in 2 cases. Root resorption of adjacent teeth was noted in 3 of 6 cases where information existed. Computed tomography was thought to be quite valuable in determining the nature and extent of the tumour. Although an elevation of serum alkaline phosphatase was observed in our case, results of laboratory tests were mostly of no diagnostic significance. Surgical removal was employed in 22 cases alone or in conjunction with irradiation and/or chemotherapy. Of 14 cases on whom information was available, local recurrence occurred in 6 cases in which radiotherapy was not given and distant metastasis in 2 of 10 cases on whom information was available. Of 20 patients on whom information was available on the postoperative course, 7 patients died 5 months to 6 years after the primary treatment. PMID- 3049680 TI - Bulimia: an entity in search of definition. AB - A review of research literature that addresses personality correlates of bulimia revealed a major problem which affects that research and its interpretation: Criteria used to define this disorder have not been consistent across investigators. A related issue is that this disorder has been described by a variety of names, such as dietary chaos syndrome, binge-eating syndrome, bulimia nervosa, etc. The second major problem that contributes to the uncertainty about the personality of bulimics is methodology. Among these have been the inclusion of individuals with a history of anorexia nervosa, the use of researcher-specific self-reports and questionnaires (primarily within a narrow population), and a lack of consistency in the testing measures used to assess personality. This very basic issue needs to be resolved if the findings in bulimia research are to be cumulative and generalizable across studies. Suggestions are offered as to how this resolution should occur. PMID- 3049681 TI - Some comments concerning the status of research activities of clinical psychologists in the Veterans Administration. PMID- 3049675 TI - Insulin receptor function in fibroblasts from patients with leprechaunism. Differential alterations in binding, autophosphorylation, kinase activity, and receptor-mediated internalization. AB - Insulin receptor function was examined in cultured skin fibroblasts from three patients with leprechaunism (Ark-1, Minn-1, and Can-1), a rare syndrome of severe insulin resistance and neonatal growth retardation. All three patients cell lines demonstrated insulin binding less than 15% of control. This was primarily due to reduced affinity of the receptor in Can-1 and due to reduced number of receptors in the other two cell lines (Ark-1 and Minn-1). When expressed as a fraction of total insulin bound, the percentage of cell-associated insulin internalized and degraded did not differ between the patient cell lines and the controls. However, chloroquine, which inhibited degradation by 50% in the control cells, had no effect in the cells from the patients. When normalized to insulin binding, insulin receptor autophosphorylation was normal in cells from Can-1, but reduced in those of Ark-1 and Minn-1. In contrast, the receptor-associated tyrosine kinase activity toward exogenous substrates was decreased in all three patient cell lines. These results suggest that leprechaunism is a biochemically heterogenous disease associated with a variety of alterations in receptor function. Cells from Ark-1 and Minn-1 exhibit parallel alterations in receptor autophosphorylation and kinase activity. Cells from Can-1 demonstrate normal receptor autophosphorylation but reduced kinase activity, thus displaying a unique form of a mutant insulin receptor. Despite reduced kinase activity, all three cell lines exhibit normal rates of insulin internalization, but decreased lysosomal-mediated degradation. Our data imply that receptor autophosphorylation and kinase activity may be regulated separately and that kinase activity may be linked to insulin degradation, but not necessarily internalization. PMID- 3049682 TI - Silver deposition in the cervix after application of silver nitrate as a cauterising agent. AB - Large amounts of metallic silver pigment were found in 10 cervical biopsy specimens taken for the histological grading of cervical intraepithelial neoplasia (CIN). The assessment of CIN was made much harder because the pigment obscured the morphological detail of the epithelial cells, and in some it was very difficult to determine whether koilocytosis or CIN I was present. The silver can easily be removed by a simple chemical method. PMID- 3049684 TI - Evaluation of the Microbact-24E bacterial identification system. AB - The Microbact 24E (MB24E) system is a commercial microsystem for the identification of common clinical isolates of Enterobacteriaceae and non fermenting Gram negative bacilli, and consists of dehydrated substrates distributed in the wells of microtitre trays. This system was compared with the API20E for the identification of 386 bacterial isolates, which included 284 clinical and 102 environmental organisms. There was 97% and 91% agreement for the identification of clinical isolates of Enterobacteriaceae and other Gram negative bacilli, respectively. The identification of environmental isolates by both systems was less satisfactory. The API20E has a more extensive database than the MB24E and is thus more reliable for the identification of rare or unusual organisms, but the MB24E is cheaper, is easy and convenient to use, and is suitable for a routine microbiology laboratory. PMID- 3049685 TI - Vero toxin producing E coli in haemorrhagic colitis. PMID- 3049686 TI - Speech and hearing science in ancient India--a review of Sanskrit literature. PMID- 3049683 TI - Glycol methacrylate (GMA) embedding for light microscopy. II. Immunohistochemical analysis of semithin sections of undecalcified marrow cores. AB - A routine method allows bone marrow biopsy specimens to be embedded in glycol methacrylate (GMA), a water miscible plastic, and to benefit from the advantages of good morphology with immunoperoxidase detection of a wide range of cellular antigens useful in diagnosing and classifying various haematopoietic disorders. Marrow cores were fixed in cold Bouin's solution, rinsed in cold phosphate buffer, dehydrated in cold methanol, infiltrated and embedded in cold GMA, then polymerised at 4 degrees C. Sections were cut at 2 micron thickness with a Tungsten carbide knife in a Jung's high performance microtome (Autocut). Antigenecity was preserved when drying slides at room temperature but pronase digestion was necessary to re-expose the antigens in bone marrow biopsy sections embedded in GMA. Histostik, a new adhesive, was used to coat the glass slides to prevent section loss during enzyme digestion and immunostaining procedures. This method of adapting plastic embedding to undecalcified marrow cores preserves marrow architecture and cellular details and it can serve as a useful adjunct to analyse the bone marrow from patients with myeloproliferative and lymphoproliferative disorders. This technique may also be applicable in non haematological malignant conditions which affect the marrow. PMID- 3049687 TI - Biology of hypopigmentation. AB - A review of the basics of pigment cell biology is followed by a discussion of the characteristics of several disorders of hypopigmentation. By determining such features as inheritance pattern, time of onset (congenital, childhood, adulthood), natural history (stable vs progressive), type of pigment loss (diffuse or circumscribed), distribution of lesions (generalized vs localized), degree of pigment loss (incomplete or complete), number of melanocytes, if any, in biopsy specimens of affected areas, type of melanocytic dysfunction, and associated inflammation or infection, one can classify the disorders of hypopigmentation. The proposed pathophysiology for each disorder of hypomelanosis is presented. PMID- 3049688 TI - Reticulohistiocytoma of the dorsum. AB - To clarify the nature of reticulohistiocytoma of the dorsum, 19 cases, including three of the seven original cases described by Crosti, were evaluated clinically, histologically, and immunologically. In seven cases gene rearrangement analysis was also performed. Results indicate that reticulohistiocytoma of the dorsum must be considered a primary cutaneous B cell lymphoma of follicular center cell origin. This localized skin disease has a very slowly progressive course, with many patients showing no systemic involvement even after prolonged follow-up. PMID- 3049689 TI - Amoxicillin and diaper dermatitis. AB - Multiple skin sites and the gastrointestinal tract of 57 infants with otitis media were cultured quantitatively for Candida albicans before and after antibiotic therapy. Ten days of systemic therapy with amoxicillin was associated with a twofold increase in the recovery of C. albicans from the rectum and skin. Infants who developed diaper dermatitis had a significant increase in the number of C. albicans organisms recovered from these sites. We conclude that the use of amoxicillin increases the risk for developing diaper dermatitis. PMID- 3049691 TI - Why is flushing limited to a mostly facial cutaneous distribution? AB - Despite the systemic nature of many agents that provoke flushing reactions, the erythema is most prominent in the "blush area." To elucidate the physiologic basis for such a limited distribution, two types of flushing challenges were studied in normal volunteers. Nicotinic acid provokes flushing through a direct action of vasodilator prostaglandins on vascular smooth muscle. The flushing reaction provoked by oral thermal challenge is mediated via neural mechanisms. Both agents led to increases in cutaneous blood flow at both malar and forearm sites. Both absolute and proportional increases were consistent with the view that the greater vascular capacitance in the visible, superficial cutaneous vasculature in the blush area accounts for the limited distribution of flushing in response to a systemic stimulus. PMID- 3049692 TI - Chancroid: clinical variants and other findings from an epidemic in Dallas County, 1986-1987. AB - Dallas, Texas, recently joined the ranks of cities in the United States that have been plagued by outbreaks of chancroid. This article describes an epidemic of chancroid in Dallas County and presents cases that illustrate the spectrum of clinical illness that this venereal disease can produce. Also reviewed are the current diagnostic and therapeutic methods of managing patients with chancroid. PMID- 3049690 TI - Pemphigoid and ulcerative colitis. AB - We report eight patients with ulcerative colitis who have subsequently developed pemphigoid. Four patients were investigated to determine the pattern of pemphigoid antibody staining on whole and chemically split skin and oral mucosa to distinguish between pemphigoid and epidermolysis bullosa acquisita. The findings were suggestive of pemphigoid with a predominantly epidermal pattern. Three patients were studied for the presence of antibodies against the colon (which were absent). The relationship between pemphigoid and ulcerative colitis is discussed in relation to a case-control study. PMID- 3049693 TI - Should the patient with generalized pruritus be evaluated for malignancy? PMID- 3049695 TI - In memory of Franz Herrmann on his 90th birthday, Aug. 2, 1988. PMID- 3049694 TI - Bullous pemphigoid of childhood: report of a case and immunoelectron microscopic studies. PMID- 3049697 TI - Study of irritant contact dermatitis produced by repeat patch test with sodium lauryl sulfate and assessed by visual methods, transepidermal water loss, and laser Doppler velocimetry. AB - Eleven subjects received patch tests with 2% sodium lauryl sulfate on a total of 34 anatomic sites. The irritant effect was monitored by visual means, laser Doppler velocimetry, and measurement of transepidermal water loss. After 1 week, repeat patch tests with 2% sodium lauryl sulfate were performed on the same site, and the effect was monitored as before. Although the skin had returned to normal or near normal before the repeat patch test, an augmented response to irritation was generally seen after the repeat patch test, particularly in transepidermal water loss, which showed an augmented response in 29 of the 34 anatomic sites. The clinical implications regarding the healing of contact irritant dermatitis are discussed. PMID- 3049696 TI - Confirmation of early Kaposi's sarcoma by polyclonal antibody to type IV collagen. AB - By the use of polyclonal antibody to type IV collagen that binds to epitope(s) retained in routinely processed tissues, we have confirmed the presence of early Kaposi's sarcoma in skin lesions that were not diagnostic on histologic examination. This reagent is of considerable use, particularly in cases of rapidly developing Kaposi's sarcoma with acquired immunodeficiency syndrome in which histologic features may not be fully developed. PMID- 3049700 TI - Dyshidrosiform pemphigoid. PMID- 3049698 TI - Chemical warfare. Cutaneous lesions from mustard gas. AB - Cutaneous lesions of eight patients evacuated from the Iran-Iraq war zone are reported. The patients were exposed to mustard gas, and there was a spectrum of dermatologic lesions. In the mild cases only there was a sunburnlike erythema, but in the most severe cases this erythema was followed by large and disseminated bullae, which histologically showed a subepidermal location with full-thickness epidermal necrosis. The resorption of these blisters was followed by hyperpigmentation, which was more evident in the intertriginous areas. In these last cases there also were ocular, respiratory, and gastrointestinal manifestations. In the skin this mustard gas seems to act through a primary irritant mechanism and without allergic contact sensitization. PMID- 3049699 TI - Perilesional linear atrophy and hypopigmentation after intralesional corticosteroid therapy. Report of two cases and review of the literature. AB - We report on the cases of two patients in whom linear perilesional hypopigmentation and atrophy developed after intralesional injection of corticosteroids for treatment of keloids. Evaluation of our patients and the previously reported cases showed that perilesional linear atrophy or hypopigmentation (or both) is a distinct complication after intralesional or intraarticular administration of corticosteroids and is probably due to lymphogenous spread of the steroid suspension. PMID- 3049701 TI - Primary eruption of psoriasis in a split skin graft. PMID- 3049703 TI - Effectiveness of relaxation and visualization techniques as an adjunct to phototherapy and photochemotherapy of psoriasis. PMID- 3049704 TI - Unusual manifestations of syphilis with human immunodeficiency virus infection. PMID- 3049702 TI - Successful treatment of lipohypertrophic insulin lipodystrophy with liposuction surgery. PMID- 3049705 TI - Idiotypes, anti-idiotypes, and autoimmunity. AB - Over the past two decades it has become clear that the ability of a host to generate antibodies against a wide variety of potential antigens is due to structural diversity in the antibody molecule within the variable region. This diversity results in sites within the molecule that are themselves immunogenic. These immunogenic sites are called idiotopes, and the collection of idiotopes on a single antibody molecule determines that antibody's idiotype. The idiotype of an antibody molecule is defined serologically by a second antibody termed an anti idiotype. Anti-idiotypic antibodies can recognize antibody molecules bearing similar or identical structures within the variable regions, which are often on or near sites of antigen binding. Investigation into the nature of idiotype and anti-idiotype interactions has increased our knowledge of antibody structure, antigen-antibody interactions, the regulation of antibody production, and the nature of autoimmune disorders. This review will discuss the nature of idiotypes and anti-idiotypes and their potential role in the diagnosis, management, and treatment of autoimmune, infectious, and malignant diseases. PMID- 3049706 TI - Recollections of dermatology in the 1930s. PMID- 3049707 TI - Personal experiences with Stephen Rothman, M.D. PMID- 3049709 TI - Microcystic adenoma of the pancreas: spectrum of computed tomographic findings. AB - Fourteen cases of microcystic adenoma (serous cystadenoma) of the pancreas were reviewed and radiological findings were correlated with pathological specimens. Microcystic adenomas appeared grossly either as solid tumors with innumerable tiny cysts or as honeycombed cystic tumors depending on the size and number of cysts and amount of connective tissue. Dynamic enhanced CT of the tumor reflected the amount of connective tissue and appeared as (a) densely enhanced spongy masses (n = 6: classic appearance); (b) cystic masses with (n = 6) or without (n = 1) enhanced septa; or (c) dense diffusely enhanced mass (n = 1). Ultrasound similarly showed a variety of features such as echogenic masses with or without small cystic portions, multilocular cysts, or mixed hyperechoic and hypoechoic masses. The ultrasonic features mainly reflected the dominant sizes of cysts. Angiography almost always showed inhomogeneously hypervascular masses with tumor vessels. The imaging diagnosis is easy and conclusive in classic subtypes, but a correct diagnosis can be made even in other subtypes. However, mucinous cystic neoplasm can be confused with microcystic adenoma with large cysts and a small amount of connective tissue, and islet cell tumors can be mistaken for microcystic tumors with minute cysts. PMID- 3049708 TI - Seasonal acclimation of bank voles and wood mice: nonshivering thermogenesis and thermogenic properties of brown adipose tissue mitochondria. AB - Seasonal acclimation of nonshivering thermogenesis and brown adipose tissue was studied in wild bank voles (Clethrionomys glareolus), yellow necked field mice and wood mice (Apodemus flavicollis, A. sylvaticus). Both, voles and mice increased their capacity for nonshivering thermogenesis during winter. Thermogenic properties of brown fat (cytochrome c oxidase activity, mitochondrial protein content, GDP-binding of brown fat mitochondria) showed similar changes during seasonal acclimation; Clethrionomys and Apodemus spp. both showed lowest thermogenic properties in the summer during August, a rapid increase during fall, and highest levels of thermogenic activity in the winter months. With regard to changes in body weight and brown fat mass these species show different strategies for seasonal acclimation. In Clethrionomys a reduction of body mass in the winter was found, both in the wild population as well as in individual animals housed in the laboratory. A. flavicollis showed a reduction of body weight during fall, whereas A. sylvaticus maintained a constant body mass throughout the year. Brown fat mass and cellularity increased in the Apodemus spp. during winter, in parallel with the thermogenic properties of brown fat, whereas in Clethrionomys brown fat mass and cellularity remained seasonally constant. These species live in the same habitat and were trapped in the same area. It is concluded that seasonal improvements of in vivo and in vitro thermogenesis are very similar in these species, although the physiological basis for this improvement is different in Clethrionomys and Apodemus. PMID- 3049710 TI - Persistent primitive trigeminal artery aneurysm evaluated by MR imaging and angiography. AB - The primitive trigeminal artery is the most common of the persistent carotid basilar anastomoses with an angiographic incidence of 0.6%. Fifteen cases of aneurysms of a persistent primitive trigeminal artery have been reported in the literature. A 16th patient is presented in whom an incidental trigeminal artery aneurysm and a thrombosed aneurysm of the vertebrobasilar system are evaluated by magnetic resonance imaging and angiography. PMID- 3049711 TI - Neonatal craniopharyngioma: CT and ultrasonographic features. AB - A case of craniopharyngioma in a neonate is reported and the literature on neonatal craniopharyngioma reviewed. First reported in 1952 and previously considered rare, the diagnosis of these neonatal tumors has increased with the use of noninvasive imaging including CT and ultrasound. The CT and ultrasound features of this uncommon but potentially treatable entity are described. PMID- 3049712 TI - The attached gingiva in children: diagnostic, developmental and orthodontic considerations for its treatment. AB - In the clinical context, the width of the attached gingiva is established by substracting the sulcus depth from the width of the keratinized gingiva. Summarizing all factors, it becomes clear that control of marginal inflammation appears to be the most important key to limiting--and possibly reversing--an otherwise progressive recession. A nonsurgical approach should be the clinician's first choice. PMID- 3049713 TI - Anomalies of form and number, fused primary teeth, a correlation of the dentitions. AB - Fusion is rare in the primary dentition (range .14 percent to 3 percent). Patients with fused primary lateral incisors and canines have about a 75 percent chance of lacking the succedaneous lateral incisor. Patients with fused incisors have less than a 20 percent chance of having a missing permanent tooth. PMID- 3049715 TI - The prevalence of dental attrition and its association with factors of age, gender, occlusion, and TMJ symptomatology. AB - Dental attrition severity in 222 young adults was assessed from dental casts as the sum of the most severe facet in each arch segment. The attrition scores were compared by age, gender, bruxism awareness, prior bite adjustment, orthodontic class, maxillomandibular relationship, and temporomandibular dysfunction symptoms. Awareness of bruxism was not associated with the wear scores and should not be used to define bruxist groups. Attrition scores did not differ significantly between age groups, indicating that notable attrition, when present, often occurs early. Men had higher attrition scores than women (p less than 0.01), despite fewer signs and symptoms. Dental attrition was not associated with the presence or absence of TMJ clicking, TMJ tenderness, or masticatory muscle tenderness. Class II division 2 males had laterotrusive attrition scores lower than those of Class III (p less than 0.05). Class III females had lower incisor attrition scores than did other Angle Classes (p less than 0.05). Discernible dental attrition in a non-patient population was not associated with signs and symptoms of temporomandibular disorders, nor with the occlusal factors studied. These results are compatible with the findings in other studies that point to bruxism as a centrally induced phenomenon common to all people and unrelated to local factors. PMID- 3049716 TI - An in vitro evaluation of the prevention of caries on overdenture abutments. AB - Teeth prepared as overdenture abutments are susceptible to caries, and it has been shown that brushing by itself is not sufficient to prevent this process. This study evaluated the preventive effect of a remineralization gel which has a low fluoride concentration and compared its effects with those of a phosphate fluoride gel (Karigel), which has a much higher concentration of fluoride. Twenty extracted anterior teeth from patients aged 50 to 70 years were prepared as for overdenture abutments. Each tooth was sectioned into three fragments. An acidified gel system was used to produce artificial caries lesions on the occlusal and root surfaces of each fragment. One fragment of each tooth was treated with the remineralizing gel, the second fragment with a high-fluoride gel, and the third fragment served as the control. Ten teeth were removed at two weeks and again at four weeks, and were sectioned and prepared for histological examination. The depth of the lesions was measured from standardized photomicrographs by means of a sonic digitizer. The conclusions were: (1) Lesions on the occlusal tended to be deeper than those on the root surfaces at four weeks but not at two weeks; and (2) the high-fluoride gel was more protective than the low-fluoride remineralizing solution at both time periods on the occlusal but not on the root surface. PMID- 3049714 TI - A complete fusion in the primary human dentition: a histological approach. AB - A trace of cementum totally surrounded by dentine could not be found here; this histological evaluation gave no further information about the origin of this double tooth. The fact that the dental pulp was complex could not be observed radiographically. PMID- 3049717 TI - An in vitro evaluation of artificial caries-like lesions on restored overdenture abutments. AB - An acidified dialyzed gelatin gel system was used to determine the caries resistance of a variety of restorative materials used to obturate the canal orifice of overdenture abutment teeth. The restorative materials used were Tytin, Tytin + Copalite, P30 + Scotchbond, Fuji Ionomer-Type II, and Miracle Mix. Polarized light microscopy and microradiography were used to examine the caries like lesions adjacent to the restorations. The lesions formed in the Fuji Ionomer Type II and Miracle Mix groups appeared arrested at the wall adjacent to the restoration, and did not penetrate apically down the wall as did those associated with the other restorative materials. The mean depths of lesions adjacent to Fuji Ionomer-Type II and Miracle Mix restorations were significantly less than those of Tytin, Tytin + Copalite, or P30 + Scotchbond. PMID- 3049718 TI - Microleakage in class II composite restorations bonded to dentin using thermal and load cycling. AB - The effectiveness of a dentin-bonding agent (DBA) and a glass-ionomer cement liner (GIC) in reducing the marginal microleakage at the cervical margin of Class II posterior composite restorations was examined. The effect of load cycling on microleakage of this type of restoration was evaluated. The Class II cavities were prepared in extracted human pre-molar teeth, with the cervical margins finished approximately 1 mm below the CEJ. Combinations of DBA, GIC, and control (no liner) were used as mediators between resin and dentin. All restorations were placed incrementally with a 40-second cure per increment and finished with high speed diamond burs. Half the samples of each treatment group received cyclical axial loading and half did not. All samples were thermocycled at 5 degrees and 55 degrees C for 500 cycles of one min each, stained (four hr) in 50% AgNO3, and sectioned following stain development. Microleakage was scored linearly along the dentin-restoration interface. All restorations exhibited microleakage which was unaffected by load cycling. Both the DBA and the GIC significantly reduced microleakage independently. When GIC was present, the additional presence of the DBA did not provide a statistically significant additional reduction of microleakage. PMID- 3049719 TI - The effect of varying diagnostic thresholds upon clinical caries data for a low prevalence group. AB - This study investigated the impact of employing differing diagnostic thresholds on clinical caries data in studies of groups with low caries prevalence. Data from clinical examinations of 287 Hong Kong dental students were analyzed by means of the CARIES microcomputer software package. This software allows for re calculation of raw data according to three different diagnostic thresholds (D3, D2, and D1). When "enamel" and "initial" lesions (as defined by WHO criteria) were included in the calculation of DMFT, its value increased from 3.0 (D3) to 5.9 (D1), while the percentage of individuals considered "caries-free" decreased from 28.2% to 7.0%. Little change was found in the magnitude of the intra examiner reproducibility, when calculated at each threshold, for a random 10% of the subjects. It was unfortunately not possible to calculate inter-examiner reproducibility in this study. The use of criteria which might be misinterpreted as being similar, but which use differing effective diagnostic thresholds, can dramatically influence the reported level of dental caries. In view of these findings, it may be necessary for the question of diagnostic thresholds to be re examined and to receive greater emphasis in future studies. PMID- 3049720 TI - Fracture mechanics parameters for failure prediction of composite resins. AB - This study contains the first part of a research project in which the applicability of fracture mechanics parameters to predict failure of a restored tooth was investigated. Fracture mechanics parameters have been used in dental research before, but were restricted to comparative studies between various brands of composites. The critical values of the opening mode stress intensity factor (KI), its equivalents, the strain energy release rate (GI), and the J integral (JI), were measured with single-edge notched-bend (SENB) specimens of dental composite in a three-point bend test. The measured values of KIc for Silux (KIc = 0.99 +/- 0.03 MNm-3/2) and P-30 (KIc = 1.88 +/- 0.12 MNm-3/2), compared with values from the literature, show quantitative agreement. The J integral was computed by means of finite element analysis (FEA) on a two-dimensional model of the SENB specimens. The critical value of the J integral (measured with SENB specimens, notch depth-to-width ratio (a/W) = 1/2) was used to predict failure of specimens having an arbitrary geometry. In this study, failure was predicted for SENB specimens with notch depth-to-width ratio (a/W) = 1/4 and 3/4. The predicted deflection and load at failure correspond well with the measured deflection and load. PMID- 3049722 TI - Determining safe limits for untransfused, outpatient liposuction: personal experience and review of the literature. AB - Limiting liposuction volumes to avoid transfusion is sound surgical practice. Although the plastic surgery literature reports frequent use of transfusions in liposuction surgery, dermatologists almost never use blood replacement after liposuction. Techniques which favor less bleeding include sufficient use of fresh epinephrine, cryoanesthesia, use of smaller cannulas, fluid preloading, proper preoperative evaluation, serial liposuction, intramuscular steroids, and rapid application of pressure garments. A review of the literature and personal experience are detailed. PMID- 3049721 TI - DICOR surface treatments for enhanced bonding. AB - Treatments for preparing castable ceramic surfaces for enhanced bonding to specially formulated resin-based cements were examined. An ammonium bifluoride etch combined with gamma-methacryloxypropyl-trimethoxysilane produced shear bond strengths higher than when an ammonium bifluoride treatment was used alone. The method of curing the silane was highly significant in the contribution to the cement/substrate bond strength, with the heat-cure producing the highest values. Long-term water storage tests indicated that the cement bond with etch plus silane-treated castable ceramic surfaces (whether heat or chemically cured silane was used) demonstrated no significant decrease in strength after a one-year period. PMID- 3049723 TI - Selection and utilization of liposuction cannulas. AB - The liposuction surgeon can now choose from a wide variety of cannulas. However, the author has found that most procedures can be successfully performed using five or six basic types. Types of cannulas and techniques of liposuctioning are briefly described. PMID- 3049724 TI - A case control clinical trail of two wound drainage collection systems. PMID- 3049725 TI - Assessment tools for the identification of patients at risk for the development of pressure sores: a review. PMID- 3049726 TI - A clinical report on the comparison of a drain/tube attachment device with conventional suture methods in securing percutaneous tubes and drains. PMID- 3049727 TI - Polarization assay studies of human neutrophil motility. PMID- 3049728 TI - Administration of IgG fraction of epidermolysis bullosa acquisita (EBA) serum into mice. PMID- 3049729 TI - Autoantibodies to vimentin-type intermediate filament in patients with progressive systemic sclerosis. PMID- 3049730 TI - A trial of acupuncture for progressive systemic sclerosis. PMID- 3049731 TI - Case report and study of collagen metabolism in Ehlers-Danlos syndrome type II. PMID- 3049732 TI - Desmoplastic malignant melanoma--an electron microscopic and immunohistochemical study of a case. PMID- 3049733 TI - Malignant melanoma in Malaysia--a report of 23 cases. PMID- 3049734 TI - Pigmented neurofibroma. PMID- 3049736 TI - Sarcoid reactions in a patient with congenital syphilis. PMID- 3049735 TI - Squamous cell carcinoma developing in thermal keratosis. PMID- 3049737 TI - The effect of etretinate on plasma fibronectin in patients with vulgar psoriasis. PMID- 3049738 TI - Trace element analyses in pseudoxanthoma elasticum. PMID- 3049740 TI - Instrumentation and physical factors related to visualization of stenotic and regurgitant jets by Doppler color flow mapping. AB - Major clinical uses of the new Doppler color flow mapping technologies involve the imaging of disturbed flow through cardiac defects or valves. Nevertheless, there is little general understanding of the determinants of flow and of how flow is imaged by these new systems. This review will attempt to relate the hydrodynamics through a simplified stenotic or regurgitant orifice with the physics and sampling theories relevant to the functioning of Doppler color flow mapping systems. The goal will be to characterize the velocity resolution, spatial resolution, sensitivity and performance of these systems so that clinicians can understand why flow looks the way it does on Doppler color studies and which aspects of flow mapping can be expected to become more quantifiable than they are at present. PMID- 3049739 TI - Maintaining ethical standards in today's dental practice. A historical perspective. PMID- 3049741 TI - Morbid events and risk factors for death after cardiac transplantation. PMID- 3049742 TI - Honorary fellowship conferred on David W. Talmage, March 1988. David W. Talmage, MD (1919-). PMID- 3049743 TI - Tryptase levels in nasal-lavage fluid as an indicator of the immediate allergic response. AB - To examine mast cell involvement in allergic rhinitis, levels of tryptase, a specific marker for mast cell activation, and histamine, a marker of mast cell and basophil activation, were measured in nasal-lavage fluid after nasal-allergen challenge. Twelve atopic subjects with allergic rhinitis and five nonatopic subjects were challenged with timothy grass or ragweed pollen at increasing doses of allergen. Tryptase and histamine levels were determined by an ELISA and radioenzyme assay, respectively; clinical responses were measured by assessment of sneezing, rhinorrhea, nasal congestion, and ocular tearing or itching. A positive clinical response was observed in seven of the atopic subjects and in none of the nonatopic subjects. Tryptase levels increased at least sevenfold higher than baseline levels in 100% of the atopic clinical responders and reached a maximum at the same dose of allergen where clinical symptoms were maximal. In contrast, histamine levels were only threefold or greater elevated in five of seven atopic clinical responders at this dose of allergen. (Histamine levels were lower in one subject and were only 50% higher in another subject than the corresponding baseline value.) Histamine levels and symptom scores were maximal at the same dose of allergen in only four of seven clinical responders. Overlap of peak mediator levels in subjects without a clinical response with those of the clinical responders occurred only in the case of histamine. Tryptase levels in nasal-lavage fluid appear promising as a useful indicator of allergic reactions and indicate that mast cell activation is the major factor in the immediate nasal allergic response. PMID- 3049744 TI - Cat- or dog-induced immediate and late asthmatic responses before and after immunotherapy. AB - To assess the effect of specific immunotherapy (IT) on the immediate and late asthmatic responses induced by cat or dog extract bronchial provocation testing, we studied seven cat-sensitive and six dog-sensitive subjects with asthma. Among the cat-sensitive subjects, two of three subjects displaying only an immediate asthmatic response before IT lost the response after 3 months of IT with cat extract, whereas only one of four subjects with the dual (immediate and late) asthmatic responses had ablation of such responses (no significance for the one tailed test). Three subjects had worsening of the immediate response after IT. Among dog-sensitive subjects with asthma, three of five subjects with only an immediate response demonstrated an ablation of the immediate response, whereas the one subject who was the only subject with a dual response before IT lost the dual response after dog-extract IT (p value of 0.005 on one-tailed test). Immediate systemic side effects, including one episode of anaphylaxis, occurred on seven occasions of 326 injections with cat extract compared to only one episode of rhinitis of 289 injections with dog extract. PMID- 3049746 TI - Techniques for accessing the medical literature. I. Currently available approaches. AB - Today physicians and scientists have a wide range of choices in their quest to locate both clinical and scientific information. This article describes the range of choices available to allergists and immunologists as they attempt to locate information on a subject and offers some simple suggestions for managing the increased body of biomedical information. PMID- 3049747 TI - Recognition of pollen and other particulate aeroantigens by immunoblot microscopy. AB - Most grass-pollen types appear identical by normal light microscopy. Restricted antigenic cross-reactivity of Cynodon dactylon (Bermuda grass) pollen allowed development of a new method to identify antigens associated with grass-pollen grains. Pollen applied to the surface of an adhesive tape was blotted onto nitrocellulose, and the blots were identified by anti-Bermuda-grass antibodies, second antibody, and fluorescence microscopy. Of the 44 species of grass pollen studied for specificity of the method, the only species to demonstrate uniformly bright staining were Cynodon dactylon, Elymus triticoides, Elymus cinereus, and Koeleria cristata. Thirty-one species were negative, and nine other species demonstrated occasional brightly fluorescent spots, suggesting contamination. The immunoblotting method was used to study Tucson air collected continuously by a Burkard pollen and spore trap throughout April 1986. Each 2-hour transect of the adhesive tape from the trap was examined by immunoblotting and by normal light microscopy to compare antigen particle counts with grass-pollen counts. The mean antigen-particle concentration, 52.8/m3 of air, was higher than the mean grass pollen concentration, 21.9/m3 of air, suggesting presence of amorphous Bermuda grass antigens in air samples. Antigen-particle concentration, not grass-pollen concentration, correlated significantly with wind velocity, temperature, and time of day. PMID- 3049748 TI - Individual dietary intake methodology: a 50-year review of progress. AB - This article provides an overview of five decades (1936 through 1987) of publications on individual dietary assessment methodology, such as dietary histories, estimated and weighed food records, food frequency questionnaires, and 24-hour recalls. Representative studies were selected to characterize data collection and analyses methods of each decade. During the 1930s and 1940s, dietary intake methodology was in its initial stages; popular methods were the dietary history technique and lengthy food records. The 1950s were characterized by extensive comparisons of methodologies, which now often included shorter-term food records and 24-hour recalls. The 1960s ushered in large scale epidemiological studies, the food frequency technique, and use of computer technology for computation; the 24-hour recall was still widely used in that decade and the next. Advances of the 1970s and 1980s include expansion of nutrient databases, sophisticated statistical techniques for analysis, and refinement of data collection methodologies for analysis, and refinement of data collection methodologies. The chronological approach used in this review not only highlights progress of each decade but also identifies the repetitive efforts of some studies. The need for creative approaches is emphasized as current research needs are identified. PMID- 3049745 TI - Proceedings of the Task Force on Guidelines for Standardizing Old and New Technologies Used for the Diagnosis and Treatment of Allergic Diseases. Washington, DC. June 18-19, 1987. PMID- 3049749 TI - Elders' nonadherence, its assessment, and computer assisted instruction for medication recall training. AB - This study investigates three questions related to the problem of medication nonadherence among elders. First, does recall failure play a significant role in nonadherence? Recent research suggests that it may not. Second, can the new portable bar code scanner technology be used to study nonadherence? Other forms of monitoring are obtrusive or inaccurate. Finally, can inexpensive computer assisted instructions (CAI) be used to teach mnemonic techniques specifically designed to improve medication schedule recall? Current research on memory training teaches nonspecific mnemonics and uses the expensive classroom approach. Results of the present study suggest that physically active and cognitively alert elders do have significant nonadherence (control group = 32.0%) problems related to forgetting and that CAI courseware can significantly reduce (medication recall training group = 10.0%) this form of nonadherence. Portable bar code technology proved easy to use by elderly patients and provided detailed information about the type of forgetting underlying nonadherence. Most significant recall failure was in the complete forgetting to take medication rather than delays in medicating or overmedicating. PMID- 3049751 TI - Physiology and complications of bed rest. AB - Prolonged bed rest causes major cardiovascular, respiratory, musculoskeletal, and neuropsychological changes. Complications often result from these effects of bed rest, especially when aging has already decreased the reserves in these systems. These problems can be limited by judicious prescription of modified bed rest. PMID- 3049750 TI - Clinical decision-making in catastrophic situations: the relevance of age. Report of a conference convened by the American Geriatrics Society, April 17-18, 1987. PMID- 3049753 TI - Clinical comparison of the threshold-related and single-intensity strategies of the Humphrey Field Analyzer. AB - The threshold-related, eccentricity-compensated strategy was compared to the single-intensity strategy using a minimal test point distribution. Forty-two eyes known to be free of any visual field defect and 44 eyes with documented defects were tested with each screening strategy. The threshold-related, eccentricity compensated strategy yielded a slightly poorer sensitivity (84.4%) and specificity (97.6%) than the single-intensity sensitivity (90.9%) and specificity (100%). An explanation for this decreased sensitivity is offered. The 40-point test was useful for some general screening. Recommendations are made for appropriate utilization and interpretation. PMID- 3049752 TI - Public financing of Medicare. PMID- 3049754 TI - Optometry: a legal history. PMID- 3049755 TI - The four psychologies of psychoanalysis and their place in clinical work. AB - Clinical psychoanalysis has led to the development of four conceptually separable perspectives on the functioning of the human mind. These are referred to herein as the psychologies of drive, ego, object relations, and self. Their clinical relevance is explored by applying them to issues regarding evenly hovering attention and the mutative factors in psychoanalysis. Those two areas, in turn, are seen to undergo an expansion when viewed from the perspective of each of the four psychologies. PMID- 3049756 TI - Freud's three theories of love in the light of later developments. AB - Evidence is presented that Freud developed three very different theories on love. These theories were not integrated into a coherent theory. In subsequent developments each theory had its own history. The paper discusses the history of the controversy on the genital character, the relation between love and gender identity, between love and narcissism, the hierarchical structure of the capacity to love, and the relation between love and object loss. The impact of some concepts such as symbiosis and the rapprochmente subphase on the understanding of conflicts in loving is raised. While at present differences in emphasis make it difficult to build a coherent psychoanalytic theory of love, it is productive to bring divergent views in touch with each other. A unified theory of love based on psychoanalytic observations is suggested. PMID- 3049757 TI - Mutagenic activity of acetonitrile and fumaronitrile in three short term assays with special reference to autoinduction. AB - Two aliphatic nitriles, acetonitrile and fumaronitrile were tested for their genotoxic potential in three mutagenicity test systems: the Salmonella/microsome assay, an assay using Saccharomyces cerevisiae (strain D7), and the bone marrow micronucleus test. Both compounds were tested with and without metabolic activation in the yeast and the bacterial test systems using S9 preparations from phenobarbitone-pretreated and autoinduced rats. Autoinduction was performed by chronic (7 days) application of a dose equivalent to a 5% oral LD50-value of the respective compound. With yeast strain D7 both nitriles induced low levels of gene conversion in the presence of phenobarbitone-induced liver homogenate. An increase in the number of ile+-revertants was not detectable under any condition. Neither of the compounds showed mutagenic activity in the Ames test with or without metabolic activation. A weak positive effect of acetonitrile could be detected in the micronucleus test 24 h after i.p.-injection of the compound using a dose of 60% LD50. Fumaronitrile showed positive results with a 50% LD50 dose 48 h after administration to mice not preinduced. After 1 week of autoinduction these effects did not appear anymore, with the exception of acetonitrile 72 h after application of a dose amounting to 60% of the oral LD50-value. PMID- 3049758 TI - Double labeling of the paravertebral sympathetic C system in the bullfrog with antisera to LHRH and NPY. AB - The specificity of synaptic contacts between pre- and postganglionic cells in the sympathetic C system has been examined by immunocytochemical localization of two neuropeptides. Sections of bullfrog paravertebral sympathetic ganglia were stained with antibodies to luteinizing hormone releasing hormone (LHRH) and neuropeptide Y (NPY). Preganglionic synaptic boutons containing LHRH immunoreactivity were found to make contact with a subpopulation of postganglionic cell bodies and with some clusters of small intensely fluorescent (SIF) cells. In ganglia 9 and 10, 95.8% of the neurons contacted by LHRH containing boutons were also positive for NPY-like immunoreactivity and conversely, 99.3% of the neurons that contained NPY-like immunoreactivity were contacted by LHRH-containing boutons. Qualitatively similar results were found in most other paravertebral ganglia. These observations support the conclusions that preganglionic C axons selectively innervate C-type ganglion cells and that virtually all C-type ganglion cells and some SIF cells receive a direct LHRH input. Moreover, they suggest that a pattern of specific connections between two sets of peptidergic neurons is expressed throughout most of the paravertebral sympathetic chain of the bullfrog. PMID- 3049759 TI - Spectral analysis of heart rate variability in the assessment of autonomic diabetic neuropathy. AB - We studied heart rate variability in 49 uncomplicated diabetics (27 with insulin therapy; 22 with oral hypoglycemic agents) and in 40 age-matched controls. An automatic autoregressive algorithm was used to compute the power spectral density (PSD) of beat by beat RR variability derived from the surface ECG. The PSD contains two major components (a low frequency approximately 0.1 Hz (LF) and a high frequency, respiratory linked, approximately 0.25 Hz (HF] that provide, respectively, quantitative markers of sympathetic and vagal modulatory activities and of their balance. As compared to controls, in diabetics, besides a reduced RR variance at rest (2722 +/- 300 and 1436 +/- 241 ms2, respectively), we observed during passive tilt an altered response of spectral indices of sympathetic activation and vagal withdrawal, suggestive of a complex modification in the neural control activities. In addition, we compared this approach to the commonly used clinical tests score, and observed that the latter provides overall results similar to those obtained with spectral changes induced by tilt (r = 0.42; P less than 0.01). Of potential clinical importance is that the data obtained with spectral analysis appear more thoroughly quantifiable and do not require the active collaboration of the patients. PMID- 3049761 TI - [Histological study of 4 lenses from epikeratoplasty. Anatomo-clinical correlation]. AB - Four removed kerato-lens lenticles (1 for keratoconus, 1 for aphakia, and 2 for myopia) have been studied histologically. The reasons for removal of the lenticles were poor visual acuity in spite of a satisfactory anatomic result in three cases and epithelialization of the interface in one case. The epithelium presented modifications, particularly with exoserosis of basal cells and irregularities of thickness. The Bowman membrane was irregular and interrupted in some parts. The corneal stroma had few cells; the keratocytes were observed in the periphery of the lenticle and near the interface. PMID- 3049760 TI - [A trial use of lenses of human placental collagen in epikeratoplasty procedures]. AB - Epikeratoplasty is a refractive surgery procedure described by Werblin and Kaufman which is simple, effective and reversible. However its development is hampered by the need of human corneas. We evaluated this procedure in rabbits and monkeys using lenticules of human collagen. The collagen is obtained from frozen human placentas, washed and subjected to pepsin digestion; it is contained in the supernatant of centrifugation and can be isolated by a physico-chemical treatment. 15 days after surgery the lenses of 14 rabbits were completely covered with a layer of epithelial cells. 6 months after surgery, the eyes of 14 monkeys were quiet and the lenses were covered with an epithelium made up of 2 to 5 layers of cells. The lens exhibited no inflammatory infiltration and there was no inflammatory reaction around the lens. These studies are still preliminary and longer term tests are necessary. These initial results are very encouraging, prompting us to develop the use of this type of material in refractive surgery. PMID- 3049762 TI - [Surgery of ptosis]. PMID- 3049763 TI - [A new case of Cohen's syndrome]. AB - We report a detailed description of ocular alterations, especially uveal coloboma, as shown in a patient with Cohen's syndrome. We also report new lesions, not previously described in the revised bibliography: lens and optic head colobomas, hyaloid artery and anterior "vasculosa lentis" tunica vestige. The most characteristic ocular alterations previously described in the bibliography are eyelid antimongoloid fissures, strabismus and myopia. PMID- 3049764 TI - [Diabetic maculopathy and its treatment]. PMID- 3049765 TI - [Adjustable surgery in strabismus]. AB - Detailed description of the technique of adjustable sutures on the medial rectus muscle. Discussion of the indications and complications. PMID- 3049767 TI - [Metabolism of the lens crystalline and cataract]. PMID- 3049766 TI - [Bone choristoma of the bilateral choroid and asymmetric microphthalmia]. AB - The authors report a case of associated bilateral asymmetrical microphthalmia and osseous choristoma of the choroid in a seventeen year old young woman. This association, to the author's knowledge, has not been previously reported in the literature. The bone choristoma was present in the eye's posterior poles and the clinical features were studied by electroretinography, fluorescein angiography, A B scan ultrasonography, computerized axial tomography and nuclear magnetic resonance. The authors hypothesize that a choroid developmental tumor was produced by exposure to the rubella virus during embryonic life. PMID- 3049769 TI - On the relationship of adult daughters to their mothers. PMID- 3049770 TI - Frontiers of adult development in theory and practice. PMID- 3049768 TI - Effect of a new long-acting somatostatin analogue (SMS 201-995) on glycemic and hormonal profiles in insulin-treated type II diabetic patients. AB - Five type II diabetic patients were studied after secondary failure of oral agents, with and without the addition of the new long-acting somatostatin analogue SMS 201-995 to an intermediate-acting insulin regimen. SMS 201-995 was administered twice daily, before breakfast and dinner, as 100 micrograms sc injections, and resulted in a lowering of plasma glucose, as well as of plasma glucagon and serum C-peptide levels. SMS 201-995 abolished postprandial glycemic and xylosemic peaks related to meals and to oral d-xylose when they were taken shortly after the administration of the analogue, while it had no effect on glycemic and xylosemic increments that followed the midday meal. The new somatostatin analogue improves glucose tolerance in type II diabetic patients, both by inhibiting counterregulatory hormones and by delaying and reducing intestinal absorption of nutrients. Its administration could lead to a reduction of daily insulin requirements. Our findings indicate that SMS 201-995 may have a role as an adjunct to insulin in the management of type II diabetic patients after secondary failure of oral agents. PMID- 3049771 TI - Age as a basis for allocating lifesaving medical resources: an ethical analysis. AB - In light of the growing prominence of an age criterion in patient selection, it is essential to scrutinize the ethical legitimacy of arguments being offered both for and against using age as a criterion. Accordingly, the present study first explores the primary justifications for an age criterion, then examines the criterion's weaknesses. Weaknesses are grouped into two areas: deficiencies in the justifications of the criterion, and overarching critiques. Finally, a way forward in the midst of the present controversy is suggested. The study's conclusion is that an age criterion per se is unjustified, though age may play a carefully defined role in medical assessments relevant to patient selection. PMID- 3049772 TI - Labor union involvement in occupational safety and health, 1957-1987. AB - Right-to-know policies and related market-oriented occupational health policies require an institutionalized means through which workers can interpret and act on information about quality differences among jobs. In principle, labor unions could play this role. However, union coverage has been declining since the 1950s, and the decline has accelerated in recent years. This paper documents the growth in occupational health and safety activities in unionized workplaces from 1957 to 1987 and the decline in union representation in hazardous workplaces from 1971 to 1986. It also analyzes the relationship between right-to-know and right-to-refuse hazardous-work guarantees under industrial relations and occupational health law. PMID- 3049773 TI - The role of substitutes in policy analysis: acute care services in state Medicaid programs. AB - Using four acute care equations (inpatient, physician, outpatient, and clinic) from a larger model of Medicaid, this research examines the "contents" of policy outcomes. This closer examination of outcomes brings to light the interactions between redistributive programs and services and the role of substitutes and complements in state-level policy analysis. (A substitute is a benefit or service that can be used instead of another to produce a similar outcome; a complement is a benefit or service that is likely to result in the use of another benefit or service.) Support is found for the inclusion of these theoretical constructs in policy analysis. Regarding Medicaid, the author concludes that physician, outpatient, and clinic services all complement hospital services; that physician and outpatient services substitute for one another; that state AFDC and SSI policy decisions have a greater impact on utilization than Medicaid-specific eligibility and service decisions do; and that the factors driving utilization (supply, demand, etc.) vary dramatically across acute care settings. The implications for Medicaid policymaking are also discussed. PMID- 3049774 TI - Home blood transfusions: the medical, economic, and legal issues surrounding a new treatment procedure. AB - Blood transfusions have almost always been confined to hospital settings in the past. Recent medical care trends have shifted some therapies (e.g., renal dialysis, hemophilia treatment) into the patient's home. Transfusions are now being given in increasing numbers to stable patients in their homes. This paper examines the medical aspects, the economic issues, and the legal implications of such transfusions. Candidates for home transfusion must be carefully chosen primarily according to the medical guidelines for such treatment. Any legal issues must be satisfactorily answered before approval is given. The paper concludes that if done properly, home transfusions can be safe, cost-effective, and convenient for a carefully selected segment of patients. PMID- 3049776 TI - Sickle-cell anemia--a review. AB - Sickle-cell anemia is a common disease, affecting more than 50,000 blacks in the United States. Since 1970 the morbidity and mortality have improved, with patients surviving well into their fourth decade. This article discusses the spectrum of serious complications of sickle-cell anemia. Crises and complications that result from the sickling process are presented with recommendations for emergency department evaluation and management. The painful bone crisis and pain control are also discussed. PMID- 3049775 TI - Cervical injury in head trauma. AB - Criteria for excluding cervical spine injury in patients who have sustained blunt head or neck trauma were prospectively studied at four hospitals in the Chicago area. The authors attempted to define a subset of these adult patients who, based on clinical criteria, could reliably be excluded from cervical spine radiography, thus avoiding unnecessary radiation and saving considerable time and money in their evaluation. Patients fell into four groups: (1) patients who were awake, alert, and had no complaint of neck pain or tenderness on physical examination; (2) patients who were awake, alert, but had complaint of neck pain or tenderness on physical examination laterally over the trapezius muscle only; (3) patients who were awake, alert, but had complaint of central neck pain or tenderness on physical examination over the cervical spine or center of the neck; and (4) patients who were not fully awake or alert, were clinically intoxicated, had other painful or distracting injuries, or had focal neurologic findings. Patients in group 4 had significantly more fractures (21/387) when compared with all other patients (7/478). Patients with central neck pain or tenderness (group 3) had significantly more fractures (7/237) than patients without pain or tenderness or with these findings limited to the trapezius area (0/236). It is clear that patients who have altered mental status, abnormal examination findings, distracting injury, or pain or tenderness over the cervical spine must have cervical spine radiographs.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3049778 TI - Third Consensus Conference on HIV Testing. PMID- 3049779 TI - Report of the Third Consensus Conference on HIV testing sponsored by the Association of State and Territorial Public Health Laboratory Directors. PMID- 3049777 TI - Methylene chloride poisoning: a paradigmatic review. AB - The incidence of reported cases of toxicity resulting from methylene chloride exposure has increased within the last decade. A vast majority of these reports involve acute episodes, and the prevalence of domestic poisoning is relatively high. Diverse pathologic sequelae attributed to methylene chloride or its metabolites have been reported, although a distinct bias for central nervous system effects is evident. paradoxically, detoxification of methylene chloride via the mixed-function oxidase pathway is an inherently intoxicating event. Although the anomalous conversion of methylene chloride into carbon monoxide has increased the popular awareness of methylene chloride poisoning among medical personnel, lack of experience in diagnosis and treatment of methylene chloride poisoning is widespread. This review of 26 cases spanning 50 years reveals that the industrial and domestic use of methylene chloride is equally widespread. A compendium of the clinical experience with methylene chloride poisoning is presented. PMID- 3049780 TI - Application of HIV methods to the detection of other retroviruses. PMID- 3049781 TI - Handwashing, Semmelweis, and chlorine. PMID- 3049782 TI - Receiver operator characteristic (ROC) curves. AB - Receiver operator characteristic curves describe the trade-off between appropriate diagnoses of true disease and inappropriate positive diagnoses in healthy persons. ROC curves may be used to compare the accuracy of two or more tests, diagnostic algorithms, or the performance of diagnosticians. PMID- 3049783 TI - Pseudomeningitis--another nosocomial headache. PMID- 3049784 TI - Do antibiotic combinations prevent the emergence of resistant organisms? PMID- 3049785 TI - Medical museum notes. PMID- 3049786 TI - Electronic information for physicians: a new dimension in solving problems. PMID- 3049787 TI - Medical museum notes. PMID- 3049788 TI - Brain abscess: a review. PMID- 3049789 TI - The hyperglycemic hyperosmolar syndrome. PMID- 3049790 TI - A new lectin-gold complex for ultrastructural localization of galacturonic acids. AB - We report the development of a cytochemical affinity technique for detection of galacturonic acids at the ultrastructural level. The highly purified gonad lectin from Aplysia depilans (AGL) was tagged with colloidal gold particles and used for labeling carbohydrates in resin-embedded sections of various plant and fungal tissues. Patterns of AGL binding sites were compared to those obtained with a D galactose-specific lectin, Ricinus communis agglutinin I. Differences in labeling patterns were noted, indicating that the lectins exhibited differential carbohydrate binding. In addition, the considerable loss of labeling over isolated wheat coleoptile walls treated for removal of pectin, after incubation with the AGL-gold complex, strongly suggested an affinity of AGL for pectic substances. A series of cytochemical controls, including sugar inhibition tests, has proven the specificity of the technique and the high affinity of AGL towards galacturonic acids. The potential value of this new lectin for ultrastructural studies on cell wall pectic substances in plant biology and pathology is demonstrated. PMID- 3049793 TI - Bibliography of the current world literature in hypertension. PMID- 3049791 TI - Procollagen intermediates during tendon fibrillogenesis. AB - The purpose of this study was to correlate ultrastructural features of tendon collagen fibrils at various stages of development with the presence of procollagen, pN-collagen, pC-collagen, and the free amino propeptides and carboxyl propeptide of type I procollagen. Tendons from 10-, 14-, and 18-day chicken embryos reveal small, well-defined intercellular compartments containing collagen fibrils with diameters showing a unimodal distribution. At 21 days (hatching) and 9 days (post hatching) and at 5 weeks (post hatching), the compartments are larger, less well-defined, and there is multimodal distribution of tendon fibril diameters. Procollagen and the intermediates pN-collagen and pC collagen are present in tendons up to 18 days. Thereafter there is a marked reduction in procollagen, whereas the intermediates persist throughout all stages of development. Similarly, free amino propeptides and carboxyl propeptides of type I procollagen were found at all stages. The amino propeptide of type III procollagen was restricted to the peritendineum until 7 weeks post hatching. At that time, a network of fibrils containing the amino propeptide of type III procollagen was seen delineating well-circumscribed compartments of collagen fibrils throughout the entire tendon. This study supports the notion that pN- and pC-collagen have an extracellular role and participate in collagen fibrillogenesis. PMID- 3049792 TI - Lanthanide chelates as new fluorochrome labels for cytochemistry. AB - Anti-rabbit IgG labeled with a new fluorescent europium chelate was used to localize rabbit IgG to human smooth muscle myosin in a histological section. The antibody labeled with the europium chelate could be viewed with a conventional fluorescence microscope with a steady-state light source. This result encourages the development of a time-resolved fluorescence microscope, because a significant improvement in the signal-to-noise ratio can be anticipated. PMID- 3049795 TI - Seventy-five years of immunology: the view from the MHC. PMID- 3049794 TI - Cyclosporine A and hypertension. PMID- 3049796 TI - Differential radiosensitivity among B cell subpopulations. AB - We have previously shown that low doses of ionizing radiation selectively impair a functionally defined B cell subpopulation. Normal mice, after exposure to 200 rad of ionizing radiation, have normal or near normal splenic plaque-forming cell responses to thymus-independent type 1 Ag, but reduced responses to thymus independent type 2 Ag. Here, we confirm and extend the original findings by using hapten-specific serum RIA to demonstrate this differential radiosensitivity is systemic. We also examined splenocytes stained with a panel of lymphocyte surface Ag by FACS analysis to determine if these functional changes are accompanied by a physical alteration of the B cell pool of irradiated mice. Single-parameter FACS analyses demonstrate a diminution in both B cell number and the heterogeneity of membrane Ag expression within the surviving B cell pool after irradiation. In contrast, T cells are relatively radioresistant as the relative percentage of T cells in the irradiated splenocyte pool increases, whereas the heterogeneity of membrane Ag expression remains constant. Multiparameter FACS analyses indicate that B cells with the sIgM much greater than sIgD phenotype are more radiosensitive than B cells of the sIgM much less than sIgD phenotype. In addition, immunohistochemical analysis of splenic sections stained with anti-IgM or anti-IgD reveal the enhanced radiosensitivity of marginal zone B cells. PMID- 3049797 TI - T cell antigen receptors in autoimmunity. AB - Three mAb to variable region determinants of the alpha/beta-chain TCR were used to detect discrete populations of peripheral blood T cells. T cells sharing a TCR determinant defined by such an antibody presumably use the same or similar TCR V or J genes for their alpha- or beta-chains. Thus analysis with these mAb provides a tool to investigate TCR gene usage and expression. Since autoantigen specific T cells may play an important role in initiating autoimmune diseases, TCR were analyzed in different autoimmune diseases and control groups including rheumatoid arthritis, Graves disease, idiopathic thrombocytopenic purpura, psoriasis, SLE, insulin-dependent diabetes mellitus, and in nonautoimmune control diseases and normals. Purified T cells were stained by indirect immunofluorescence with three mAb to TCR variable regions: mAb S511 stains 1.8 +/- 0.9% (mean +/- 2 SD), mAb C37 stains 3.4 +/- 1.5% and mAb OT145 stains from 0 to 6% of T cells from normal donors. Several individuals were identified with expanded subsets of positive T cells. One patient with adult ITP followed during a 12-mo period consistently had elevated percentages of T cells staining with the mAb OT145 (15.9 to 24.5%). These cells were found to be exclusively CD8+. By Southern blotting DNA prepared from these OT145+, CD8+ cells, but not DNA from the patient's OT145- T cells, revealed a clonal rearrangement using a beta-chain C region probe. Thus this patient had a monoclonal expansion of CD8+, OT145+ cells. Hyperexpression of a TCR variable region, as defined by the available mAb, could not be associated with any of the diseases studied. Examination of T cells at the site of autoimmunity, such as T cells from rheumatoid arthritis synovial fluid, revealed normal percentages of cells staining with these mAb. Immunoperoxidase staining of psoriatic lesional skin showed no striking enrichment of T cells bearing one or the other TCR type. PMID- 3049798 TI - Structure-activity studies of human IL-1 beta with mature and truncated proteins expressed in Escherichia coli. AB - IL-1 beta is synthesized as an inactive 31-kDa intracellular protein, which is then processed upon secretion to an active 17-kDa carboxyl-terminal fragment. To identify the minimal portion of IL-1 beta required for activity, we constructed several deletion mutants of mature IL-1 beta. These included three amino-terminal deletions of 10, 16, and 81 amino acids, two carboxyl-terminal deletions of 17 and 72 amino acids, and one internal fragment between amino acids 17 and 81. Expression of the mutants was monitored by Western blots and immunoprecipitation. With one exception, all of these mutants and the full length 17-kDa IL-1 beta were expressed as soluble protein in Escherichia coli and could be assayed for activity and receptor binding in lysates without further purification. Whereas the intact 17-kDa IL-1 beta retained full biologic activity (greater than 10(7) U/ml of lysate) and competed for binding with 125I-labeled IL-1 beta, none of the lysates containing IL-1 beta deletion mutant proteins had activity or competed for binding to receptor at significantly higher concentrations. The loss of function in the smallest C-terminal deletion mutant does not appear to be due to the direct involvement of these C-terminal residues in receptor binding because both monoclonal and polyclonal antisera directed to this region bind to IL-1 beta but do not neutralize its activity. Therefore, this region is probably indirectly involved in sustaining the structure of the receptor-binding site. PMID- 3049799 TI - Plasmacytoma growth factor activity of murine granulocyte-macrophage colony stimulating factor. AB - Mouse plasmacytoma FLOPC21 was adapted to culture in the presence of a mouse Th cell supernatant. A stable factor-dependent cell line was derived from this culture and the factor responsible for its growth was identified as granulocyte macrophage colony-stimulating factor. PMID- 3049800 TI - Idiotypic vaccination as a treatment for a B cell lymphoma. AB - To develop a model for the active immunotherapy of human B cell malignancy we vaccinated tumor-bearing animals with a well defined tumor associated Ag, the idiotypic Ig. The tumor used was the mouse B cell lymphoma BCL1, a highly malignant tumor in which transfer of a single tumor cell to a syngeneic mouse is capable of causing disease and eventual death. Varying doses (10(2) to 10(4] of BCL1 cells were given to mice on day 0 of the experiment, and treatment by active immunization was initiated on day 3. Immunization with purified, tumor-derived, idiotypic IgM (BCL1 IgM) coupled to keyhole limpet hemacyanin (KLH) was highly effective in treating mice challenged with 10(2) or 10(3) BCL1 cells, but less effective in mice that had received 10(4) tumor cells. Immunization with unconjugated BCL1 IgM showed no signficant therapeutic benefit. Coupling of the IgM to KLH led to higher levels of anti-idiotypic antibody after immunization; however, the higher levels were probably not responsible for the control of the malignancy as there was no correlation in healthy immunized animals between the levels of anti-idiotypic antibody, measured immediately before tumor challenge, and survival. This lack of correlation is due to the emergence of variant tumors in such protected mice. A more significant factor in the therapeutic advantage of KLH conjugation could be that immunization with BCL1 IgM-KLH led to an earlier induction of the anti-idiotypic response than immunization with BCL1 IgM and, as the BCL1, lymphoma divides rapidly, the speed of induction of the immune response may be important in outstripping tumor cell growth. Mice with BCL1 tumour showed some evidence of immunosuppression as indicated by a reduced ability to mount an immune response against KLH. Although it is not possible to model spontaneous human lymphoma accurately, the generation of a functional anti-idiotypic response capable o eliminating a malignant animal lymphoma in situ opens up the possibility of a limited trial of active immunotherapy in selected human patients. PMID- 3049801 TI - Development of donor-derived thymic lymphomas after allogeneic bone marrow transplantation in AKR/J mice. AB - The transplantation of bone marrow cells from BALB/c (but not C57BL/6 and C3H/HeN) mice was observed to lead to the development of thymic lymphomas (leukemias) in AKR/J mice. Two leukemic cell lines, CAK1.3 and CAK4.4, were established from the primary culture of two thymic lymphoma, and surface phenotypes of these cell lines found to be H-2d and Thy-1.2+, indicating that these lymphoma cells are derived from BALB/c donor bone marrow cells. Further analyses of surface markers revealed that CAK1.3 is L3T4+ Lyt2+ IL2R-, whereas CAK4.4 is L3T4- Lyt2- IL2R+. Both CAK1.3 and CAK4.4 were transplantable into BALB/c but not AKR/J mice, further indicating that these cells are of BALB/c bone marrow donor origin. The cells were found to produce XC+-ecotropic viruses, but xenotropic and mink cell focus-forming viruses were undetectable. Inasmuch as thymic lymphomas are derived from bone marrow cells of leukemia-resistant BALB/c strain of mice under the allogeneic environment of leukemia-prone AKR/J mice, this animal model may serve as a useful tool not only for the analysis of leukemic relapse after bone marrow transplantation but also for elucidation of the mechanism of leukemogenesis. PMID- 3049802 TI - Bone marrow cell transplants involving intra-H-2 recombinant inbred mouse strains. Evidence that hemopoietic histocompatibility-1 (Hh-1) genes are distinct from H-2D or H-2L. AB - Hemopoietic histocompatibility (Hh) Ag are noncodominantly expressed on bone marrow stem cells and other normal and neoplastic cells of hemopoietic origin. H 2/Hh-1 allogeneic or parental-strain bone marrow grafts are eliminated in a determinant specific manner by NK cells. In inbred mouse strains, seven Hh-1 alleles representing combinations of five different Hh-1 antigenic determinants are described. Each Hh-1 allele maps in the vicinity of H-2D, and the genes that map to Hh-1 are transacting regulatory genes. The expression of a particular determinant depends on the absence of the regulatory gene and the presence of the appropriate structural gene. The primary focus of this study is to ascertain whether the Hh-1 phenotype and the serologic H-2DL typing are always correlated or whether recombinant can separate the two. To achieve this, we used a panel of irradiated hosts that are able to recognize the different Hh-1 determinants on the bone marrow cells of congenic intra-H-2 recombinant donors. We report: 1) the majority of strains show a correlation between Hh-1 and H-2DL: 2) B10.RQDB and B10.WB strains dissociate Hh-1 from Lb: 3) nine H-2S/D interval recombinant strains exhibit no correlation between the H-2DL type and Hh-1 phenotype; and 4) in two strains from this group, B10.D2 (R106) and B10.RSF5, H-2S/D crossovers occurred within Hh-1r, (Hh-1 regulatory). We conclude that Hh-1r is a distinct regulatory locus mapping telomeric of H-2S and centromeric of, although probably closer to, H-2D. PMID- 3049803 TI - Locally administered monoclonal antibodies to lymphocyte function-associated antigen 1 and to L3T4 prevent cutaneous graft-vs-host disease. AB - In vivo treatment with mAb directed against T cell surface molecules has been shown to prevent or reverse graft-vs-host disease (GVHD) and experimental autoimmune diseases, where T cells play a critical role. In this study we investigated the effects of in vivo administration of anti-I-A, anti-L3T4, and anti-lymphocyte function-associated Ag 1 (LFA-1) mAb on the development of murine cutaneous GVHD and delayed-type hypersensitivity responses evoked by local transfer of class II-MHC Ag-restricted, cloned autoreactive T cells. Prevention of cutaneous GVHD was achieved with local administration of either anti-L3T4 or anti-LFA-1 mAb, but not with anti-I-A mAb, while delayed-type hypersensitivity responses were inhibited by all the mAb. These results indicate that an I-A Ag independent mechanism may be also operative in the development of the cutaneous GVHD, unlike the delayed-type hypersensitivity responses. Anti-LFA-1 mAb appeared to be more potent at inhibiting cutaneous GVHD rather than delayed-type hypersensitivity responses, whereas anti-L3T4 mAb inhibited equally both the responses. These results suggest that LFA-1 molecule may be involved in the epidermal invasion of T cells. Treatment with anti-LFA-1 mAb was found to be still, although less, effective at preventing cutaneous GVHD, even if the mAb was administered after the development of cutaneous GVHD. PMID- 3049804 TI - Effects of T cell depletion in radiation bone marrow chimeras. I. Evidence for a donor cell population which increases allogeneic chimerism but which lacks the potential to produce GVHD. AB - The opposing problems of graft-vs-host disease (GVHD) and failure of alloengraftment present major obstacles to the application of bone marrow transplantation (BMT) across complete MHC barriers. The addition of syngeneic T cell-depleted (TCD) bone marrow (BM) to untreated fully allogeneic marrow inocula in lethally irradiated mice has been previously shown to provide protection from GVHD. We have used this model to study the effects of allogeneic T cells on levels of chimerism in recipients of mixed marrow inocula. The results indicate that T cells in allogeneic BM inocula eliminate both coadministered recipient strain and radioresistant host hematopoietic elements to produce complete allogeneic chimerism without clinical GVHD. To determine the role of GVH reactivity in this phenomenon, we performed similar studies in an F1 into parent combination, in which the genetic potential for GVHD is lacking. The presence of T cells in F1 marrow inocula led to predominant repopulation with F1 lymphocytes in such chimeras, even when coadministered with TCD-recipient-strain BM. These results imply that the ability of allogeneic BM cells removed by T cell depletion to increase levels of allochimerism may be mediated by a population which is distinct from that which produces GVHD. These results may have implications for clinical BM transplantation. PMID- 3049806 TI - Autoantibodies of primary biliary cirrhosis recognize dihydrolipoamide acetyltransferase and inhibit enzyme function. AB - Autoantibodies against mitochondria occur in the sera of patients with primary biliary cirrhosis (PBC) with characteristic reactivity to an inner membrane protein of approximately 74 kDa. To precisely define these autoantigens, we recently cloned and sequenced a rat liver cDNA (pRMIT) that encodes for all of the epitopes recognized by Ig to the 74-kDa autoantigen. In the present study we have used this recombinant probe as a tool, in addition to purified enzymes, to demonstrate by immunoblotting that the 74-kDa mitochondrial autoantigen is dihydrolipoamide acetyltransferase (EC 2.3.1.12), the core protein of the pyruvate dehydrogenase complex. Furthermore, and of particular interest, inhibition of pyruvate dehydrogenase enzyme activity was demonstrated after incubation with sera from patients with PBC but not from normal volunteers or patients with chronic active hepatitis. Such inhibition was abrogated by absorption of the PBC sera with an expressing subclone of pRMIT, designated pRMIT 603. Identification of dihydrolipoamide acetyltransferase as the target of autoimmunity in PBC provides a reagent that can be used to determine mechanisms by which this molecule is recognized. It will allow study of whether autoimmune reactivity, at the humoral or T cell level, is the basis for the pathogenesis of PBC. Additionally, such data present evidence of functional inhibition of a critical metabolic enzyme. Dihydrolipoamide acetyltransferase is well-known to mitochondrial biochemistry and, similar to identified autoantigens in other autoimmune diseases, is highly conserved in evolution. PMID- 3049809 TI - Cellular mechanisms in B lymphocyte activation and tolerance. PMID- 3049805 TI - Increased class I and class II MHC products and mRNA in kidneys of MRL-lpr/lpr mice during autoimmune nephritis and inhibition by cyclosporine. AB - The expression of MHC products in the kidneys of MRL-1pr/1pr mice was investigated. As previously described, these mice develop lupus-like nephritis with intraglomerular and peritubular Ig deposition, vasculitis, and interstitial mononuclear cell infiltration at about 12 wk of age. As the nephritis appeared, the expression of MHC class I and II products rose, as demonstrated by absorption and by specific binding of radiolabeled antibodies. Hybridization of kidney RNA with specific probes revealed an increase in specific mRNA for MHC class I and II genes and for beta2 microglobulin. Using rat monoclonals against mouse class I and II MHC products, and goat anti-rat Ig as second antibody, we showed that the increase in renal class I and II expression was localized to the basolateral membranes of tubular cells, and, in the case of class I, in arteries and glomeruli. The sites of tubular MHC expression corresponded closely to the sites of extensive peritubular Ig deposition. High doses of cyclosporine given for 6 to 8 wk reduced the peritubular Ig deposits, renal Ia and H-2K expression, and specific mRNA for beta 2-microglobulin and MHC genes, but did not reduce anti-DNA antibody levels in serum. Thus the peritubular Ig deposits and tubular MHC induction coincided in timing and location, and in their resolution with cyclosporine. The results raise the possibility that the increase in renal MHC expression not only accompanies the renal lesions, but may play a role in their pathogenesis. PMID- 3049807 TI - Temporal adaptation of neutrophil oxidative responsiveness to n-formyl-methionyl leucyl-phenylalanine. Acceleration by granulocyte-macrophage colony stimulating factor. AB - This investigation was undertaken to clarify the mechanism by which purified recombinant human granulocyte-macrophage colony stimulating factor (GM-CSF) potentiates neutrophil oxidative responses triggered by the chemotactic peptide, FMLP. Previous studies have shown that GM-CSF priming of neutrophil responses to FMLP is induced relatively slowly, requiring 90 to 120 min of incubation in vitro, is not associated with increased levels of cytoplasmic free Ca2+, but is associated with up-regulation of cell-surface FMLP receptors. We have confirmed these findings and further characterized the process of GM-CSF priming. We found that the effect of GM-CSF on neutrophil oxidative responsiveness was induced in a temperature-dependent manner and was not reversed when the cells were washed extensively to remove the growth factor before stimulation with FMLP. Extracellular Ca2+ was not required for functional enhancement by GM-CSF and GM CSF alone effected no detectable alteration in the 32P-labeled phospholipid content of neutrophils during incubation in vitro. Our data indicate that GM-CSF exerts its influence on neutrophils by accelerating a process that occurs spontaneously and results in up-regulation of both cell-surface FMLP receptors and oxidative responsiveness to FMLP. Thus, the results demonstrate that, with respect to oxidative activation, circulating endstage polymorphonuclear leukocytes are nonresponsive or hyporesponsive to FMLP; functional responsiveness increases dramatically as surface FMLP receptors are gradually deployed after the cells leave the circulation. Thus, as neutrophils mature, their responsiveness to FMLP changes in a manner which may be crucial for efficient host defense. At 37 degrees C, this process is markedly potentiated by GM-CSF. We conclude that endogenous GM-CSF, released systemically or at sites of infection and inflammation, potentially plays an important role in host defense by accelerating functional maturation of responding polymorphonuclear leukocytes. PMID- 3049808 TI - Identification of a surface antigen on Loa loa microfilariae the recognition of which correlates with the amicrofilaremic state in man. AB - Filarial infections induce a spectrum of disease in their natural hosts, and by correlating immunity found in individuals with their disease pattern, one may delineate non-pathogenic, protective mechanisms. Loa loa is causal of mild to moderate pathology, and it is unique among the human filaria in that adult worms are occasionally visible during subconjunctival migration. To study immune mechanisms controlling microfilaremia, sera from 15 subjects with amicrofilaremic occult loiasis (OL) were compared with sera from 10 subjects with microfilaremic loiasis (ML) microfilaremia, (greater than 4000/ml) for their reactions with living microfilariae (mf). An IFA was first used to detect antibodies able to bind to the surface of living L. loa mf. ML subjects either did not react (7/10) or reacted only very weakly (3/10). Highly reactive sera were found only in OL subjects; 7/15 gave very bright fluorescence, 5/15 gave moderate reactions, and 3/15 were negative. Most of these antibodies were of the IgG class. Sera from all subjects were also reacted with living mf in an antibody-dependent cellular adherence test using normal leukocytes. Sera that were strongly positive in IFA showed strong adherence and IFA-negative sera were non-reactive. To identify the Ag involved, mf were surface iodinated, detergent-extracted Ag were immunoprecipitated, and Mr was determined on SDS-PAGE. Several OL sera, all highly reactive in the above tests, precipitated a 23-kDa molecule with which all ML sea failed to react. Sera from a mandrill experimentally infected with L. loa also precipitated the 23-kDa Ag when taken post-patency. In conclusion, it appears that certain people who control L. loa microfilaremia have high levels of IgG antibodies that bind to a surface Ag of 23 kDa and are able to mediate cellular adherence. PMID- 3049810 TI - Antibody complementarity and antibody structure. PMID- 3049811 TI - The AAI, 1913 to 1988, in the first century of immunology. PMID- 3049812 TI - A century of progress: beyond molecular immunology. PMID- 3049814 TI - Booster irradiation to the spleen following total body irradiation. A new immunosuppressive approach for allogeneic bone marrow transplantation. AB - Graft rejection presents a major obstacle for transplantation of T cell-depleted bone marrow in HLA-mismatched patients. In a primate model, after conditioning exactly as for leukemia patients, it was shown that over 99% of the residual host clonable T cells are concentrated in the spleen on day 5 after completion of cytoreduction. We have now corroborated these findings in a mouse model. After 9 Gy total body irradiation (TBI), the total number of Thy-1.2+ cells in the spleen reaches a peak between days 3 and 4 after TBI. The T cell population is composed of both L3T4 (helper) and Lyt-2 (suppressor) T cells, the former being the major subpopulation. Specific booster irradiation to the spleen (5 Gy twice) on days 2 and 4 after TBI greatly enhances production of donor-type chimera after transplantation of T cell-depleted allogeneic bone marrow. Similar enhancement can be achieved by splenectomy on day 3 or 4 after TBI but not if splenectomy is performed 1 day before TBI or 1 day after TBI, strengthening the hypothesis that, after lethal TBI in mice, the remaining host T cells migrate from the periphery to the spleen. These results suggest that a delayed booster irradiation to the spleen may be beneficial as an additional immunosuppressive agent in the conditioning of leukemia patients, in order to reduce the incidence of bone marrow allograft rejection. PMID- 3049813 TI - Protein synthesis in antigen processing. AB - Recent studies indicate that Ag pass through a chloroquine-sensitive intracellular pathway in accessory cells before they are recognized by class II restricted T cells. Our results indicate that this is also true for insulin. Unexpectedly, we find that protein synthesis is required for optimal accessory cell-dependent processing of insulin and other proteins by adherent macrophages. Treatment of APC with inhibitors of protein synthesis, before and during exposure to Ag, inhibits their subsequent ability to activate murine T cell hybridomas. Experiments are described which suggest that this effect is localized to intracellular processing of Ag rather than uptake or presentation, per se. Inhibition is reversible, and is not observed in special situations where intracellular processing of Ag is not required. A distinct lag period is required for inhibition of processing after inhibition of macrophage protein synthesis. One possible interpretation is that protein synthesis is necessary for maintenance of a labile protein crucial for intracellular processing of Ag. Alternatively, the susceptibility of processing to inhibitors of protein synthesis may reflect an obligate intracellular association of Ag and newly synthesized class II histocompatibility molecules. PMID- 3049815 TI - A monoclonal antibody reacting with distinct adhesion molecules defines a transition in the functional state of the receptor CD11b/CD18 (Mac-1). AB - CD11b/CD18 (Mac-1) is a member of the leukocyte integrin family, a group of receptors that have been implicated in various effector functions and cellular collaboration in the immune response. It has been shown previously that CD11b/CD18 on cells of monocyte and myeloid lineage appears to undergo rapid activation and acquire new functional receptor specificities after exposure to selected agonists such as adenosine diphosphate (ADP). We now show that ADP induces a reconformation of the CD11b/CD18 receptor with exposure of new epitopes characteristics of this activated state. By direct binding studies, flow cytometry, and immunoprecipitation experiments, it has been found that the mAb 7E3 reacts with CD11b/CD18 only after ADP-stimulation of the cell suspension. The activated state of CD11b/CD18 induced by ADP and recognized by 7E3 can also be recapitulated by agonists inducing transients in cytosolic Ca2+ such as the chemoattractant FMLP. Moreover, this process of receptor activation does not involve quantitative mobilization of the subcellular storage pool of CD11b/CD18 to the plasma membrane. Because 7E3 also recognizes a qualitative, ADP-mediated activated state of the platelet adhesion receptor GP IIb/IIIa, it is suggested that transients in cytosolic Ca2+ might represent early secondary events for a general pathway of rapid activation of integrin receptors and, as such, represent important signals for cellular interactions in the immune response. PMID- 3049816 TI - Human monoclonal antibodies reactive with antigens of the group A Streptococcus and human heart. AB - Human mAb were produced from tonsillar or PBL of normal individuals or patients infected with group A streptococci. Lymphocytes were purified on Ficoll-Hypaque gradients and stimulated in vitro with purified group A streptococcal membranes or M protein extracts. The mAb were selected for study based on their reaction with group A streptococci, pep M5 protein, and/or M6 Escherichia coli protein. Further analysis by Western immunoblot or competitive inhibition ELISA revealed that there were two types of antibodies: one type that reacted with myosin and DNA and the other type that reacted with myosin, keratin, and/or actin. The specificities of these human mAb are similar to specificities observed in our previous studies of murine mAb reactive with group A streptococci and heart Ag. For comparison, anti-myosin antibodies were affinity purified from the sera of infected or acute rheumatic fever patients and were shown to react with myosin and DNA as well as with group A streptococci and M protein. To affinity purify these antibodies from normal sera, five times the amount of sera was required to obtain detectable quantities. These data suggest that the human mAb reactive with group A streptococci and myosin reflect the antibodies seen in sera from infected patients or acute rheumatics and that the B lymphocyte clones capable of producing these cross-reactive antibodies are also present in normal individuals. PMID- 3049817 TI - Protective immunity evoked by locally administered group A streptococcal vaccines in mice. AB - The present studies were undertaken to determine the pathogenicity of group A streptococci introduced intranasally (i.n.) into mice in an attempt to mimic mucosal infections in humans and to determine the efficacy of streptococcal vaccines administered via the mucosal route. The LD50 of type 24 streptococci (M24 strep) administered i.n. was 3 x 10(4) CFU. Throat cultures were performed in M24 strep-inoculated mice. Of 11 mice that died, 9 had positive throat cultures 3 or 4 days after i.n. challenge, and of 9 mice that survived, only 1 had a positive throat culture, indicating an association between mucosal infection and death. Postmortem examination performed on 35 mice that died after i.n. challenge showed that all had evidence of disseminated infections, and group A streptococci were recovered from the cervical lymph nodes, blood, spleen, liver, and brain. To determine vaccine efficacy, heat-killed M24 strep or pep M24 were administered i.n. to groups of mice. Whole, heat-killed streptococci and pep M24 administered locally protected mice against death from i.n. challenge infections with homologous M24 strep. The whole cell vaccine also protected against i.n. challenge infections with heterologous type 6 streptococci. Our data suggest that streptococcal vaccines administered locally evoke protective immunity against streptococcal infections. PMID- 3049818 TI - Induction of aggregation and enhancement of proliferation and IL-2 secretion in human T cells by antibodies to CD43. AB - CD43 (large sialoglycoprotein) is a heavily glycosylated protein expressed on virtually all thymus-derived lymphocytes, on a subpopulation of B cells and on granulocytes. Recently, an anti-CD43 mAb (L10) was shown to induce proliferation in T cells comparable to that induced by anti-CD3. The L10 antibody was reported to react with both sialylated and desialylated CD43. In order to further elucidate the role of CD43 in various T cell functions we have studied the biologic properties of two other mAb (B1B6 and E11B, IgG1) directed against sialic acid-dependent epitopes on CD43. Addition of low amounts of antibody (5 to 10 ng/ml) to freshly isolated T cells or to T cell lines resulted in a rapid clustering of the cells. Fab fragments were also active albeit at a 10-fold higher concentration. Aggregation was dependent on active cell metabolism (inhibited by azide and at low temperatures), on the presence of divalent cations (Mg2+) and was inhibited by antibodies to CD18 but not by antibodies to CD11a (leukocyte function-associated Ag-1 alpha). B1B6 and E11B were poorly mitogenic when added alone in soluble form to PBL or to T cells. However, supernatants from cultures of PBL treated with B1B6 for 2 days contained IL-2 activity. No increase in the number of CD25+ cells was seen during the same period. Exogenously added IL-2 did not synergize with B1B6 or E11B in activation of PBL, whereas proliferation was significantly increased by the addition of the antibodies to activation systems with low endogenous production of IL-2 (PMA or soluble anti CD3). The anti-CD43 antibodies amplified T cell proliferative responses induced by Con A or leukoagglutinin from Phaseolus vulgaris. F(ab')2 fragments enhanced proliferation significantly better than Fab fragments suggesting that cross linking of CD43 molecules was an essential features of the amplifying signal. Compared with cultures activated by Con A alone, an increased number of CD25+ cells and of blast cells as well as an increased IL-2 production was observed in cultures activated by B1B6-Con A. The results indicate that regulatory signals, which may function to modify homo- or heterotypic T cell adhesion as well as autocrine production of IL-2, can be transduced through CD43. PMID- 3049819 TI - Immunotherapy of established murine B cell lymphoma. Combination of idiotype immunization and cyclophosphamide. AB - A murine B cell lymphoma (38C13) was used to study the efficacy of idiotype immunotherapy against established tumors. Immunization of mice with 38C13 tumor derived Ig, administered after a lethal tumor inoculation, significantly prolonged survival of animals compared to control groups. The efficacy of active immunotherapy was dramatically enhanced when combined with chemotherapy. Cyclophosphamide (100 mg/kg), administered in combination with idiotype immunization to mice bearing 10-day-old, 1 to 2 cm diameter s.c. tumors, resulted in a significant prolongation of survival as compared with either cyclophosphamide or immunization alone and yielded approximately 50% cures. Additional studies combining active immunotherapy with surgical excision of the primary s.c. tumor nodule were less effective than combination chemoimmunotherapy, indicating that reduction of tumor burden was necessary, but not sufficient for effective treatment of established 38C13 lymphoma. PMID- 3049821 TI - Characterization of a method using viable human target cells as the solid phase in a cell concentration fluorescence immunoassay (CCFIA) for screening of monoclonal antibodies and hybridoma supernatants. AB - A new method has been characterized for the use of viable target cells as the solid phase for screening of hybridoma supernatants in a cell concentration fluorescence immunoassay (CCFIA). Briefly, the specific target antigen on the cells is bound by the monoclonal antibodies and revealed by use of a fluoresceinated second antibody. Separation of free from bound antibody is accomplished by filtration in the 0.2 micron filter-bottom wells of specialized assay plates. Processing is automated in a Pandex screen machine, resulting in numerical fluorescence values for each well. This method is rapid (under 1 h per 96-well plate), highly sensitive (down to 0.2 ng/ml) and sparing of target cells (0.3-2.5 X 10(4) cells per assay well). It has been applied to 37 different varieties of human solid tumor cells, as well as to human peripheral blood mononuclear cells. The cells used as targets for the characterization of this method were still capable of attachment and growth when recovered post-assay. This method was compared with a viable cell enzyme-linked immunosorbent assay (ELISA) method, showing similar sensitivity and greatly shortened assay time. Comparison of the results from this method with those obtained from flow cytometric analysis performed on viable cells showed close correlation, whereas a lower correlation was seen with immunohistochemical methods using acetone-fixed cells. Development of this method made it possible to rapidly screen many thousands of hybridoma supernatants and successfully select those which were specific for surface antigens on viable cells. PMID- 3049820 TI - Changes in gene expression associated with IFN-beta and IL-2-induced augmentation of human natural killer cell function. AB - NK function can be augmented by a variety of agents, including the cytokines IL-2 and IFN. The mechanisms associated with IL-2- and IFN-mediated augmentation of NK function are largely unknown. In order to learn more about the regulation of NK activity, we have studied changes in gene expression that occur upon treatment of a cloned line of NK cells (NK 3.3) with rIL-2 and rIFN-beta. Both IL-2 and IFN beta induced rapid augmentation of lysis mediated by NK 3.3, which was significant within 1 h, peaked at 6 h of treatment, and declined by 12 h. This enhancement of lytic function was independent of proliferation and associated with a corresponding increase in steady state levels of RNA coding for both the nuclear proto-oncogene c-myb and for the IL-2R. These changes were specific in that RNA levels of another nuclear proto-oncogene, c-myc, were increased by IL-2 but not by IFN-beta, whereas HLA class I RNA levels were relatively unchanged by either IL-2 or IFN-beta treatment. Treatment of NK 3.3 with the combination of IL 2 and IFN enhanced both lysis and c-myb expression in an additive fashion. These findings suggest that c-myb may play a regulatory role in the cytolytic activity of NK cells. PMID- 3049822 TI - Assessment of phagocytosis of plastic adherent group B streptococci by ELISA quenching. AB - An ELISA was devised to quantitate the number of group B streptococci (GBS) adherent to flat-bottomed 96-well microtiter plates. Ingestion of GBS by bovine polymorphonuclears (PMN) led to a decrease in ELISA values. This ELISA quenching proved to be proportional to phagocytosis. The method is suitable for kinetic studies of phagocytosis using glutaraldehyde to block the uptake of bacteria. The ELISA quenching test is applicable to a large number of samples and permits the use of the semi-automated equipment developed for ELISA. PMID- 3049823 TI - A comparison of immunological methods for the detection of Trichinella spiralis antigen. AB - Eight immunological methods all using the same monoclonal antibody reagent were compared for the detection of Trichinella spiralis antigen. These were based on: (1) the direct adsorption of the antigen to the immunoadsorbent (nitrocellulose membrane, polyvinyl chloride strip or microplate); (2) capture of the antigen by antibodies pre-sensitized on the immunoadsorbent; and (3) latex agglutination. The methods found suitable were: (a) capture radioimmunoassay (capture-RIA) (sensitivity: less than 0.5 microgram/ml antigen); (b) direct enzyme immunoassay (direct-ELISA) (less than 0.5 microgram/ml); (c) tube latex agglutination test (2.2 micrograms/ml); and (d) direct immunodot assay (8.8 micrograms/ml). However, the performance of the direct-ELISA was greatly affected by the presence of each of three extraneous substances (bovine serum albumin (BSA), lipopolysaccharide (LPS), normal swine muscle homogenate (NSM) added to the antigen sample. The direct immunodot assay was also affected by the presence of BSA or LPS, whereas both the capture-RIA and the tube latex agglutination methods were affected by the presence of NSM only. The findings are discussed with a view of detecting T. spiralis larvae directly from pork samples by immunological means. PMID- 3049824 TI - Kinetics of antigen-antibody reactions at solid-liquid interfaces. AB - The kinetics of antigen-antibody reactions is reviewed with special attention paid to the specific properties at solid-liquid interfaces. Theories of possible diffusion limitation in forward reaction rates are compared to experiments. It is found that the intrinsic forward reaction rate in the bimolecular antigen antibody reaction is normally not limited by diffusion either in solution or at the solid-liquid interface. However, reactions at the solid-liquid interface can be diffusion limited due to depletion of reactants close to the surface. This effect depends on geometry, intrinsic reaction rate and surface concentration of receptor molecules. Normally cell surface reactions are not diffusion limited whereas reactions at artificial surfaces often are limited by diffusion. When not limited by diffusion it is also found that the intrinsic forward and reverse reaction rates are lower for surface reactions compared to reactions in solution. Antigen-antibody reactions at solid-liquid interfaces can often be considered as practically irreversible and limited by mass transport or steric interactions. PMID- 3049825 TI - A multidot immunobinding assay for the serodiagnosis of tuberculosis. Comparison with an enzyme-linked immunosorbent assay. AB - A simple dot immunobinding (dot blot) assay procedure has been developed for the detection of antibodies directed against a soluble mycobacterial antigen preparation. This technique was compared with the widely used ELISA, in a study of samples from tuberculous patients. Dot blots were read on a densitometer. The correlation between both assays was excellent (r = 0.91; P less than 0.001); 90% of sera from tuberculous patients were detected using both techniques and a serial two-fold dilution method. Assessments of the end-points of titration curves by reflectometry and simple visual interpretation gave similar results. The dot blot assay is easier to perform and appears to be a practical alternative to ELISA for the detection of anti-mycobacterial antibodies in tuberculous patients. PMID- 3049827 TI - A radioimmunoassay for parathymosin alpha using antibodies to synthetic N terminal peptide 1-30. AB - Antibodies against the N-terminus of rat parathymosin alpha have been raised in rabbits by conjugating parathymosin alpha (1-30) to hemocyanin. A radioimmunoassay for parathymosin alpha was established by utilizing antibodies against the above polypeptide and parathymosin alpha(1-12)[Tyr] as tracer. The useful range was 5-450 pmol for parathymosin alpha. An epitope was located in the amino acid sequence 1-12. The antiserum failed to crossreact with the same molar concentrations of the partly homologous thymosin alpha 1 or prothymosin alpha. With this radioimmunoassay, parathymosin alpha was isolated from calf thymus after separation from prothymosin alpha by reversed phase HPLC. Endogenous proteases did not appear to generate N-terminal fragments of parathymosin alpha in rat liver extracts in a similar fashion to that observed for prothymosin alpha. Parathymosin alpha has a ubiquitous distribution in the human tissues examined, with levels ranging from 93 (brain) to 1043 (liver) ng of parathymosin alpha(1-30) equivalents/g (wet weight). PMID- 3049828 TI - Measurement of cutaneous blood flow velocity in delayed hypersensitivity reactions in guinea pigs. AB - The blood flow velocity in the skin of the flanks of guinea pigs can be measured precisely by laser Doppler velocimetry, provided the animal is mildly sedated. The probe holder can be attached to the shaved skin with cyanoacrylate glue and can be detached after completion of blood flow velocity measurements by application of acetone without harming the animal. The technique is pain-free and non-invasive: it can be used to make serial measurements on the same animal. The system has demonstrated hyperaemia in delayed-type responses to tuberculin PPD, ovalbumin and Keyhole Limpet Haemocyanin with immunological specificity. The time course of evolution and resolution of the vasomotor response accords with previous experience. The method can detect hyperaemia-inducing mediators in the supernatant of antigen or mitogen stimulated lymphocyte cultures and could be used for their bioassay. PMID- 3049826 TI - Isolation and characterization of highly purified streptavidin obtained in a two step purification procedure from Streptomyces avidinii grown in a synthetic medium. AB - A method is described for isolation of streptavidin from cultures of Streptomyces avidinii grown in a synthetic culture medium for 6-10 days. Streptavidin is precipitated directly from culture supernatant fluid using 80% ammonium sulfate, and the precipitate is dialyzed against water and centrifuged at 40,000 X g for 60 min. The absorbency coefficient at 280 nm of purified streptavidin was estimated to be 31.7142 +/- 0.1806 for a 1% solution. The protein appeared to be greater than 90% homogeneous by gel permeation chromatography and polyacrylamide gel electrophoresis. No biotin-binding molecules less than 70 kDa in size were detected at any step during the purification of streptavidin. Streptavidin was able to maintain a stable crosslink between two biotinylated molecules in a solid phase assay. Streptavidin purified by this method was stable in 50% glycerol/water at -20 degrees C for more than 1 year. Lyophilization or iodination did not produce apparent damage to the protein. PMID- 3049830 TI - Immunoglobulin A deposition in jejunal mucosa of children with dermatitis herpetiformis. AB - Previously we have shown by indirect immunofluorescence (IF) technique that a special IgA antibody in the sera of patients with dermatitis herpetiformis (DH) binds to the structures of the normal jejunum. Now we show by direct IF that specific IgA deposits are present in the proximal jejunum of 11/12 DH and 2/2 celiac patients before a gluten-free diet (GFD). The IgA deposition was in a tubular pattern underlying the villous and crypt epithelial basement membranes and in the lamina propria. This IgA deposition diminished or was not detectable in DH patients under a GFD for a year, and became detectable under gluten challenge in three DH patients. One patient with celiac disease and IgA deficiency, four with other intestinal diseases, and four without jejunal damage had neither jejunal IgA deposition nor circulating IgA anti-jejunal antibody. The deposition of IgA in the jejunum seemed to be correlated with the presence of IgA anti-jejunal antibody in the serum and with the presence of jejunal damage, but the degree of jejunal atrophy, the titer of the anti-jejunal antibody, and the intensity of jejunal IgA deposition in DH patient were not clearly related. Deposition of IgA in the jejunum in DH did not clearly correlate with the activity of the skin symptoms and thus may not be directly related to the pathogenesis of the skin disease of DH. PMID- 3049831 TI - Cell shedding from human plantar skin in vitro: evidence of its dependence on endogenous proteolysis. AB - Cell shedding from plantar stratum corneum was studied in vitro. Cells were shed only from the surface that had faced outwards in vivo. A quantitative measure of the cell release was obtained by determining the amount of protein that could be extracted from released and sedimented cells with 1 M sodium hydroxide. The cell release was optimal at pH 7-9 but was significant also at pH 6. The rate of cell release increased with increasing temperature, but was decreased abruptly at temperatures above 50 degrees C. The cell dissociation could be inhibited by aprotinin (Trasylol) and soybean trypsin inhibitor. Thus, it is evident that the unipolar cell dissociation in this system is mediated by an enzymatically catalyzed process, most likely with the involvement of a serine protease with an alkaline pH-optimum. The in vitro cell release shows properties indicating that it may be mediated by mechanisms also active in vivo. PMID- 3049829 TI - Paediatric endocrinology. PMID- 3049833 TI - The local regulation of blood flow evaluated simultaneously by 133-xenon washout and laser Doppler flowmetry. AB - The laser Doppler flowmeter and the 133-Xenon washout techniques of measuring cutaneous blood flow were compared for measuring the vasoconstrictor response of the hand during orthostatic maneuvres. Important discrepancies were detected for the two methods. When the hand was lowered by 40 cm a 40% decrease in blood flow was detected by the 133-Xenon method, while a 60% decrease was seen by the laser Doppler technique. Lowering the hand by 50 cm resulted in no further blood flow decrease when using the 133-Xenon method, but an 80% blood flow decrease was recorded with the laser Doppler method. A marked decrease in blood flow was recorded by the laser Doppler technique in hands that were sympathectomized or a hand that was subjected to a nerve blockade, strategies which should eliminate the orthostatic vasoconstrictor response of superficial cutaneous vessels. The 133-Xenon technique did not detect any blood flow changes in hands without sympathetic tone. We found the laser Doppler flowmetry technique unsatisfactory for measurement of blood flow changes that occur in nutritional vessels as this method measures total skin blood flow including non-capillary vessels. PMID- 3049835 TI - Characterization of human skin fibroblasts elastase activity. AB - We present evidence that enzyme activity hydrolyzing Succinoyl trialanine paranitroanilide (Suc(Ala)3NA) expressed by Human Skin Fibroblasts (HSF) in culture could be attributed to the concerted action of an endopeptidase and an aminopeptidase(s). Both endopeptidase and aminopeptidase activities were strongly inhibited by metal chelating agents and Copper and Zinc ions but were insensitive to Tissue Inhibitor of Metallo Proteases (TIMP). These protease activities coeluted on ion exchange chromatography (DEAE Tris acryl M) and were further separated by high-performance liquid chromatography HPLC (TSK 3000 SW). The endopeptidase activity, designated as HSF E1, was eluted at the position corresponding to an Mr equal to 94,000. It has only a limited elastinolytic potential as evaluated on 3H insoluble elastin, but it extensively degrades human skin elastic fibers as directly assessed on human skin tissue sections and further quantitated by automated image analysis. The level of HSF E1 increases with the number of fibroblast passages. PMID- 3049832 TI - Benzo[a]pyrene diol epoxide I modification of DNA in human skin xenografts. AB - Human skin xenografts were established on the subscapular area of skin of nude (nu/nu NIH-Swiss background) mice. When treated with benzo[a]pyrene diol epoxide I (BPDE I), specific carcinogen-DNA adducts were formed. Separation and identification of these adducts by the 32P-postlabeling technique indicated that the 7R- and 7S-BPDE I-dpGp adducts were the major adducts. Xenografts pretreated with either allantoin or anthralin showed an increase in the major 7R- and 7S BPDE I adducts compared to only BPDE I treatment. Likewise, we observed an increase in the quantity of different minor adducts. The ratios between the minor and major adducts in the pretreated grafts remained consistent with the ratio in the grafts treated with BPDE I only. We conclude that these modulators induce cells in the xenograft to enter S phase of the cell cycle. Moreover, we observed that these compounds altered the quantity of the minor carcinogen-DNA adducts without altering the overall ratios between the major 7R- and 7S-BPDE I-dpGp adducts and the minor carcinogen-DNA adducts. PMID- 3049834 TI - Prenatal diagnosis of dominant and recessive dystrophic epidermolysis bullosa: application and limitations in the use of KF-1 and LH 7:2 monoclonal antibodies and immunofluorescence mapping technique. AB - Prenatal diagnosis is now possible for junctional and recessive dystrophic forms of epidermolysis bullosa (EB); however, there is no similar published experience for dominant dystrophic EB, although data with KF-1 monoclonal antibody suggests that both forms of dystrophic EB can be identified at least postnatally with this unique probe. We now report our experience with light microscopy, electron microscopy, immunofluorescence mapping, and KF-1 and LH 7:2 monoclonal antibodies, in both a mother with dominant dystrophic EB and her fetus at risk, and in a fetus previously shown to be affected with recessive dystrophic EB. KF-1 and LH 7:2 antigens were absent in recessive dystrophic EB fetal skin, identical to findings observed postnatally. LH 7:2 was normally expressed in a mother with dominant dystrophic EB and in her fetus at risk for this disease. In contrast, while KF-1 antigen was abnormally expressed in the affected mother, it was normally expressed in only 1/7 fetal biopsies despite the fact that this fetus was shown by light and electron microscopy and immunofluorescence mapping to be unaffected with dominant dystrophic EB. We conclude that 1) transmission electron microscopy can be used to prenatally exclude the diagnosis of dominant dystrophic EB (Cockayne-Touraine variety), 2) immunofluorescence mapping is an accurate technique for prenatal as well as postnatal diagnosis of EB, and 3) KF-1 cannot by itself be used as an accurate probe for the prenatal diagnosis of dominant dystrophic EB, due to the apparent variability in the time for the normal expression of KF-1 in fetal skin during the second trimester. PMID- 3049836 TI - Implications of acquired oxacillin resistance in the management and control of Staphylococcus aureus infections. AB - Refinements in testing for resistance to penicillinase-resistant penicillins (PRP) in Staphylococcus aureus have resulted in confusion in classifying isolates as PRP susceptible or resistant. Specifically, a group of organisms has been identified that produce large amounts of beta-lactamase and appear borderline resistant. These organisms have been called "occult resistant" or "acquired oxacillin-resistant" S. aureus (AORSA). A retrospective study was conducted to evaluate the implication of this in vitro phenomenon in managing patients with AORSA infections. Among 134 patients with S. aureus infections, 89 were infected with oxacillin-susceptible S. aureus (OSSA), 26 with AORSA, and 19 with oxacillin resistant S. aureus (ORSA). There were no significant differences in outcomes when OSSA and AORSA infections were treated with PRP (chi 2MH = .990; P = .32). These results do not suppor the contention that AORSA infections should be managed differently from OSSA infections. Identifying AORSA may not be helpful in guiding antimicrobial therapy or predicting the outcome of infections with AORSA. PMID- 3049838 TI - Deletion of the Shiga toxin gene in a chlorate-resistant derivative of Shigella dysenteriae type 1 that retains virulence. AB - We used a probe specific for detecting the structural-gene sequences of Shiga toxin to analyze the genetic nature of toxin synthesis in mutant derivatives of Shigella dysenteriae type 1. A chlorate-resistant (chl) mutant (725-78) of S. dysenteriae type 1 strain 3818T, which had retained virulence but had lost production of high levels of cytotoxic activity associated with Shiga toxin synthesis, contained a complete deletion of the Shiga toxin structural-gene sequences. These structural-gene sequences were also absent in a derivative of S. dysenteriae type 1 that contained a substitution of Escherichia coli DNA in the trp region of the chromosome. Isolates of Shigella flexneri and Shigella sonnei also did not react with the probe. The low-level cytotoxic activities associated with the mutant S. dysenteriae type 1 strains or with the virulent S. flexneri and S. sonnei strains are neutralizable with antiserum to Shiga toxin; however, these cytotoxic activities are not determined by the genes encoding classic Shiga toxin. PMID- 3049837 TI - A polyclonal human IgG preparation hyperimmune for type III, group B Streptococcus: in vitro opsonophagocytic activity and efficacy in experimental models. AB - Neonates at risk for disease due to type III, group B Streptococcus (III-GBS) are those born with low serum levels of transplacentally derived, specific III-GBS antibody. Specific antibody is also required in vitro for opsonophagocytosis of III-GBS. Commercially available human immune serum globulins contain only moderate levels of III-GBS antibody. In the present study, IgG that was isolated from the serum of a human volunteer after vaccination with III-GBS polysaccharide contained very high levels of III-GBS antibody. Small amounts of this hyperimmune preparation added in vitro significantly increased the opsonophagocytosis of III GBS in neonatal sera in the presence of polymorphonuclear leukocytes. Furthermore, small volumes of the preparation were protective in mouse and neonatal rat models of III-GBS disease. The administration of such hyperimmune human IgG preparations should be considered for preventing or treating III-GBS disease in infants. PMID- 3049840 TI - Cerebrospinal fluid normalities and abnormalities in individuals infected with human immunodeficiency virus. PMID- 3049839 TI - Serodiagnosis of early Lyme disease: analysis of IgM and IgG antibody responses by using an antibody-capture enzyme immunoassay. AB - We used an antibody-capture enzyme immunoassay (EIA) to evaluate the early antibody responses to Borrelia burgdorferi in paired sera from 30 patients with erythema chronicum migrans. During acute disease, 20 (67%) patients had elevated specific IgM responses, and by convalescence (one to four weeks after treatment), 28 (93%) patients had increased IgM or IgG responses. In acute specimens, elevated IgM responses correlated with disseminated infection; however, by convalescence, most patients with either localized or disseminated disease had positive tests. Among 133 control subjects, IgM cross-reactivity was observed in 4 of 37 patients with either Epstein-Barr virus or rickettsial infections, and false-positive IgG tests were seen in 8 of 28 patients with syphilis. With antibody-capture EIA, the diagnosis of Lyme disease can be confirmed in the majority of acutely ill patients and in almost all patients by convalescence. PMID- 3049842 TI - Demonstration, by immunoelectronmicroscopy, of a cell wall antigen in Trichosporon beigelii that cross-reacts with Cryptococcus neoformans capsular polysaccharide. PMID- 3049841 TI - Early acquisition of antibody to Plasmodium falciparum sporozoites in nonimmune temporary residents of Africa. PMID- 3049843 TI - Is nonionic isotonic iohexol the contrast agent of choice for quantitative myocardial videodensitometry? AB - All currently used contrast media in coronary angiography induce a considerable hyperemic response interfering with the interpretation of circulation times derived from myocardial time-density curves. Aim of this study therefore, was to find a contrast agent with minimal hyperemic response. For this purpose 2, 4 and 6 ml of the nonionic isotonic low iodinated contrast agent iohexol (Omnipaque 140) and 6 ml of a similarly low iodinated but still hypertonic solution of the ionic diatrizoate (Urographin 30%) were administered into the left coronary artery of 8 anesthetized instrumented dogs. Heart rate was held constant by atrial pacing and left ventricular pressure, left ventricular dP/dt and mean and phasic coronary blood flow were recorded. To test the hypothesis that the hyperemic response to nonionic contrast media is partly due to an increase in inotropic state mediated by Ca++ion influx, all measurements were repeated 30 minutes after intracoronary administration of 0.5 mg verapamil. For iohexol the increase in coronary blood flow was small but significant: 12 +/- 7%, 25 +/- 11% and 38 +/- 16% for the 2, 4 and 6 ml administrations, respectively (mean +/- s.d.; p less than 0.01). For the diluted diatrizoate the increase in coronary blood flow was 65 +/- 23%. Increases for currently used contrast agents are on the order of 200-300%. After verapamil, the hyperemic response to iohexol decreased significantly to 9 +/- 5%, 20 +/- 8% and 29 +/- 12% for the 2, 4 and 6 ml administrations, respectively (p less than 0.01). The reaction to diatrizoate was not affected by verapamil. Moreover, there was a significant positive correlation between the increase in coronary blood flow and left ventricular dP/dt max under all conditions for all but one dog. We conclude that the isotonic, low iodinated nonionic contrast agent iohexol has only a moderate influence on coronary blood flow, which can be further attenuated by verapamil. By this approach, a more reliable assessment of circulation times from myocardial time-density curves obtained by digital subtraction angiography and videodensitometry becomes possible. PMID- 3049844 TI - The effect of balloon dilatation on post-stenotic myocardial perfusion before and after stimulation of coronary flow reserve: evaluation by the densitometric parameter 'mean rise time'. AB - From densitometric evaluation of digital subtraction cineangiocardiograms the parameter 'Mean Rise Time' (MRT), defined as the time from the onset of local myocardial contrast medium opacification to the point of maximal opacification can be derived; this parameter revealed a close correlation with the results on myocardial perfusion obtained by Thallium-201 scintigraphy. A prolonged 'Mean Rise Time' was indicative of an impairment of myocardial perfusion. We have developed a heart-phase gated real-time digitization procedure and computer supported method for the densitometric estimation of the MRT to obtain information about the effect of coronary balloon dilatation on myocardial perfusion before and after stimulation of coronary flow reserve by Moxaverin. In 22 patients with single vessel coronary artery disease Moxaverin caused a significant prolongation of the post-stenotic MRT (2.3 +/- 1.2s (mean +/- s.d.) vs. 2.9 +/- 1.1s, p less than 0.05), while after successful dilatation of the obstructive lesion a significant shortening of the MRT was found after stimulation of the coronary flow reserve (2.5 +/- 1.2s vs. 1.9 +/- 0.9s, p less than 0.05). A highly significant decrease in MRT after Moxaverin was measured post-dilatation in comparison to the initial pre-dilatation results (2.9 +/- 1.1s vs. 1.9 +/- 0.9s, p less than 0.005); this shows that the effect of successful balloon dilatation on the post-stenotic myocardial perfusion can be described very well by this parameter. These results demonstrate that information about post-stenotic myocardial perfusion during interventional heart catheterization can be obtained from digital densitometry. PMID- 3049845 TI - Correlations between quantitative cineangiography, coronary flow reserve measured with digital subtraction cineangiography and exercise thallium perfusion scintigraphy. AB - The goal of this investigation was to establish which anatomical parameters of stenotic lesions correlate best with its functional severity. Therefore, thirty eight patients with single vessel disease underwent coronary cineangiography and exercise/redistribution thallium-201 scintigraphy. Cross-sectional area at the site of obstruction (OA), percentage diameter stenosis (DS), the calculated pressuredrop over the stenosis (PD), as well as coronary flow reserve (CFR) derived from myocardial contrast appearance time and density were determined. The relations between CFR and the 3 anatomical parameters were described by the following equations: CFR = 4.6 - 0.053 DS, r = 0.82, SEE: 0.79, p less than 0.001 CFR = 0.5 + 0.75 OA, r = 0.87, SEE: 0.68, p less than 0.001 CFR = 3.6 - 1.5 log PD, r = 0.90, SEE: 0.62, p less than 0.001 The calculated pressuredrop was highly predictive of the thallium scintigraphic results with a sensitivity of 94% and a specificity of 90%. Therefore, the calculated pressuredrop is a better anatomical parameter for assessing the functional importance of a stenosis than percentage diameter stenosis or obstruction area. However, the 95% confidence limits of the relation between pressuredrop and coronary flow reserve are wide, making measurement of CFR a valuable addition to quantitative angiography, especially when determining the functional importance of moderately severe coronary artery lesions. PMID- 3049847 TI - Laboratory tests in the diagnosis of the chronic pancreatic diseases. Part 7. Comparison between function tests and morphological investigation in the diagnosis of pancreatic disease. PMID- 3049849 TI - A memorial biography of Pierre Desnuelle 1911-1986. PMID- 3049846 TI - Clinical application of quantitative coronary angiography using the CAAS system: preliminary results of the INTACT study (International Nifedipine Trial on Antiatherosclerotic Therapy). AB - It is the objective of the INTACT-study to test in man, whether a significant retardation of the progression of coronary artery disease is attainable with the Ca-antagonist nifedipine; this may be possible on the basis of numerous animal experiments. INTACT is a prospective, randomized, double blind, placebo controlled investigation in 423 patients with preferably early stages of coronary sclerosis in whom a progression of the disease seems likely. A proper coronary angiogram led to inclusion of the patients in the study (October 1983-June 1985). Over the following 3-years-period patients received either nifedipine 80 mg/day or placebo. The study is concluded by a control coronary angiogram with angiographic projections which are identical to those of the first coronary angiography. The extent of coronary sclerosis is objectivated by computer assisted quantitative measurement of the entire coronary arterial system with the CAAS-system (Rotterdam). For definition purposes the coronary artery system subdivided into 25 segments. Parameters for progression assessment will be mean segment diameter, minimal obstruction diameter, percentage severity of obstruction, length of obstruction and plaque area. So far 4826 coronary segments have been analyzed from the first angiograms of 383 patients. Per patient an average of 12.6 different segments could be evaluated in at least one angiographic projection. The major coronary segments could be measured in 72-93% of the patients in one or more angiographic projections (at the average about 2 different projections). Five hundred and forty-six coronary obstructions were analyzed; 131 of these were total occlusions. Only 9% of the length of the vessel contours detected by the computer algorithm required manual correction by the operators, suggesting a high reliability of the system. It can be concluded that quantitative measurement of the complete coronary artery system can indeed be obtained in a large angiographical multicenter study such as INTACT. PMID- 3049850 TI - [Comprehensive clinical study on pathogenesis of early spontaneous abortion]. AB - Clinical studies on the natural history of early spontaneous abortion included ultrasonic examination of the conceptus, determination of serum human chorionic gonadotropin beta (H), evaluation of trophoblast viability (V), chromosomal analysis of chorionic tissue and assessment of the mixed lymphocyte reaction blocking effect (BE) of serum in 22 cases with missed abortion and 4 with incomplete abortion. TV was evaluated by simultaneous staining with fluorescein diacetate-propidium iodide (FDA-PI). The data obtained were as follows: 1) Chromosomal abnormalities were observed in 13 of 21 patients analyzed. 2) V was 78.6 +/- 5.6 (n = 7) in induced abortion, 42.3 +/- 16.2 (n = 22) in missed abortion and 8.8 +/- 4.8 (n = 4) in incomplete abortion. 3) There was a significant linear correlation between the maximum diameter of the gestational sac (D) and the weight of the chorionic tissue (W) (W = 1.30 + 0.08D, r = 0.44, p less than 0.05), and between the titer of H and the weight of the viable chorionic tissue (Wv = W x V) (Wv = 0.66 + 0.01H, r = 0.77, p less than 0.01). Consequently, the following formula was devised: V = (H + 66)/(0.08D + 1.30). 4) Results obtained from the formula indicated that V stays at an elevated level for some time and then falls gradually or rapidly after fetal death in spontaneous abortion. 5) Spontaneous expulsion of the conceptus occurred when V declined to about 10% in 4 cases. 6) There was a significant linear correlation between BE and V (V = 34.58 + 0.31BE, r = 0.45, p less than 0.05). It was thus concluded that the intrauterine retention period of the conceptus after fetal death is dependent on V, which can be estimated by D, H and BE in spontaneous abortion. PMID- 3049848 TI - Lysosomal-mitochondrial interrelationships in damage to the liver in acute experimental pancreatitis in dogs. Treatment with prostacyclin (PGI2). AB - In acute pancreatitis, damage to the liver is an important aspect of multiorgan failure. In 28 dogs (20 with bile-trypsin induced acute experimental pancreatitis (AEP], 'total' and 'free' activity of lysosomal hydrolases: beta-glucuronidase, cathepsins and acid phosphatase in mitochondrial and lysosomal subfraction of the liver were determined 12 h or 24 h after the induction of AEP. The respiratory control ratio with sodium succinate as a substrate, using Clarck's electrode and uncoupler-dependent ATP-ase activity in mitochondrial subfraction, was assayed. Groups of dogs were treated or pretreated with prostacyclin (PGI2), 20 ng.kg 1.min-1 i.v. for 12 or 13 h. The relative free activity of hydrolases was significantly elevated in untreated AEP after 12 h and was partially normalized in AEP after 24 h or after 12 h followed by treatment and pretreatment with PGI2. Respiratory control ratio was twice lower than normal in AEP after 12 h and partially normalized after 24 h post PGI2 treatment. The relative free activity of lysosomal hydrolases was highly negatively correlated with respiratory control ratio. It was concluded, that during AEP in dogs the function of liver mitochondria and lysosomal stability are impaired. The significant correlation found between the mitochondrial and lysosomal lesions points to lysosomal mitochondrial interactions in liver damage in AEP. Prostacyclin in the investigated dose partially prevents the mitochondrial and lysosomal lesions in liver in this disease. PMID- 3049854 TI - Flexor tendon healing. AB - Significant changes in the concepts of tendon physiology and tendon healing have occurred in the last decade. Nevertheless, they reflect questions and controversies which date back many hundreds of years. Investigators and clinicians of today must recognise that, like their predecessors with an interest in flexor tendon surgery, they will contribute to the understanding of this very important structure, but they also will leave unanswered questions for future generations. The current interest in methods to prevent or modify adhesions is based on the presumption that the tendon can participate in the repair process. In this regard, it would be inappropriate to ignore the potential contribution by peripheral cellular elements as well. Surgeons may have moved on from the concept that pricking the tendon causes pain, twitching, and convulsions, but they have not yet clearly defined the repair process and how to consistently obtain healing of a lacerated tendon with a smooth gliding surface. PMID- 3049855 TI - The early history of contracture of the palmar fascia. Part 1: The origin of the disease: the curse of the MacCrimmons: the hand of benediction: Cline's contracture. PMID- 3049853 TI - Banked bone grafting for bone defect repair--clinical evaluation of bone union and graft incorporation. AB - One hundred and twenty-four banked bone grafts performed during the past 15 years were studied in respect to union and graft incorporation processes using X-ray, bone scintigraphy and histology. Radiological study of the chip graft showed an absence of cavity contours, and homogenization of the grafted area, followed by development of the trabecular structure. In the block graft, initial union was shown at the junctional area followed by the appearance of mottled shadows throughout the entire graft and finally differentiation of bone marrow and cortex. Bone scintigraphy showed an initial increase in RI uptake at the junction and then a gradual increase in the entire graft. Histological study showed that bone apposition on the trabeculae of the graft starts in the junction and later extends to other areas. The replacement of cancellous bone is more rapid than that of cortical bone. The porous surface seems to promote bone union and incorporation when large block grafts of cortical bone are used. PMID- 3049852 TI - [Endocrinological environment in polycystic ovarian disease (PCOD)]. PMID- 3049851 TI - [Altered expression of Lewis antigens associated with malignant transformation in endometrial tissues]. AB - Fetal, normal adult, and malignant tissues of the uterine endometrium were examined by immunoperoxidase staining for Lewis antigens. Pronounced expression of Lewis-a, Lewis-b, and Lewis-Y antigens in malignant tissues was observed, compared with that in normal adult tissues. Moreover, the amplified expression of Lewis-b antigen was considerably higher than that of Lewis-a and Lewis-Y antigens. On the other hand, Lewis-X antigen was less expressed in malignant tissues than in normal adult tissues. These results suggest that fucose transferase activity might be increased in malignant tissues and that the type I carbohydrate chain may play a role in the malignant transformation. In addition, Lewis-b and Lewis-Y antigens can be considered to be oncofetal antigens since they were frequently expressed in fetal and malignant tissues, but not in normal adult tissues. However, the functional significance of these changes in the expression of Lewis antigens remains to be investigated. PMID- 3049856 TI - Flexor tendon injuries: the results of primary repair. AB - This study is a critical analysis of results obtained following primary repair and post-operative controlled mobilisation of flexor tendon injuries which were treated by registrars with up to six months experience in hand surgery. 70 (55%) of 125 patients who underwent repair of a complete flexor digitorum profundus or flexor pollicis longus tendon injury during a 14-month period attended for review and these had a total of 140 injured digits. 93 (67%) were rated by Lister's standards as an "excellent" or "good" result. 39 (28%) occurred in "no man's land" (Zone 2) and only 19 (49%) in this area were rated "excellent" or "good". Isolated flexor digitorum superficialis tendon injuries have been excluded from this study, as have partial tendon injuries. PMID- 3049857 TI - Osteosarcoma of the carpus. PMID- 3049859 TI - The ocular distribution of bunolol in the eyes of albino and pigmented rabbits. AB - The ocular administration of a 50 microL instillation of bunolol hydrochloride, a beta 1- and beta 2- adrenoceptor blocking agent, resulted in significantly higher drug levels in the choroid/retina, iris, and ciliary body of pigmented rabbits compared with albino rabbits following topical administration. The concentrations in these tissues also persisted longer in the pigmented rabbits' eyes. However, no statistically significant differences in tissue levels were observed in the cornea or conjunctiva. The results of this study support the previously reported finding with timolol which showed longer retention of the drug in the iris, ciliary, choroid, and retina of pigmented rabbits than albinos. PMID- 3049858 TI - Hand infections with Mycobacterium chelonei: a case report and review of the literature. AB - A case is reported of soft-tissue infection in the hand due to Mycobacterium chelonei, presenting as a chronic painful swelling of a finger. PMID- 3049860 TI - Fibronectin in corneal wound healing. AB - Fibronectin is an extracellular structural protein with the unique ability to bind to both cells and collagen. It plays a major role in the development of the cornea. The universal appearance of fibronectin within 8 hours of corneal wounding has promoted major interest in its wound healing properties. The early clinical evidence for fibronectin treatment of recurrent corneal erosion and certain forms of keratitis sicca is strong. The current major problem preventing commercial use is the antigenic nature of fibronectin derived from separate species. Human pooled fibronectin has been suggested as a commercial source of this ubiquitously occurring protein. PMID- 3049861 TI - Clinical comparison of topical solutions containing trimethoprim in treating ocular surface bacterial infections. AB - Thirty-nine patients with bacterial ocular surface infections (conjunctivitis, blepharitis or blepharoconjunctivitis) were evaluated after treatment with a topically applied ophthalmic solution containing trimethoprim. A combination of trimethoprim and polymixin B (TP) produced identical clinical and microbiologic responses when compared to a solution containing, in addition to trimethoprim and polymyxin B, sulfacetamide. The sulfa component was not essential to produce desired clinical and microbiologic results. PMID- 3049862 TI - Systemic effects resulting from topical ocular administration of SCH 33861, a novel ACE inhibitor ocular hypotensive agent. AB - SCH 33861 (7- N- 1(S)-(Carboxy)-3-phenylpropyl -(S)-alanyl 1,4-dithia-7-azaspiro 4.4 nonane-8(S)-carboxylic acid) is a novel, non-sulfhydryl angiotensin converting enzyme (ACE) inhibitor with marked topical ocular hypotensive activity. The present study evaluated the degree of systemic ACE inhibition resulting from topical administration of 0.01 and 0.1% concentrations (10 and 100 fold greater than needed to lower IOP) of SCH 33861 in conscious rabbits. For reference purposes, captopril, a prototype ACE inhibitor, was examined at concentrations 5 and 20-fold greater than needed to lower IOP. Neither 0.01 nor 0.1% topical SCH 33861 led to inhibition of pressor responses to 0.3 micrograms/kg iv challenges with angiotensin I (AI) over a 4 hr period. Captopril, in contrast, caused significant attenuation of AI pressor responses. The magnitude and duration of systemic ACE inhibition following captopril administration was concentration related. Intravenous administration of the same dose administered topically did not cause any systemic ACE inhibition with 0.01% SCH 33861. Following iv 0.1% SCH 33861, 0.5 or 2.0% captopril, AI responses were inhibited 60-70% indicating the ability to inhibit ACE if any systemic absorption took place. Topical administration of 0.01% SCH 33861 twice daily for five days also did not produce systemic ACE inhibition. These findings indicate that topical amounts of SCH 33861 greatly in excess of those needed to effect reductions in IOP have an exceptionally low potential for producing systemic effects. PMID- 3049864 TI - Contribution of monocyte-macrophage system to serum alpha 1-antitrypsin. AB - The major serum antiprotease is alpha 1-antitrypsin (A1AT). Deficiency of A1AT can result in infantile cirrhosis and premature emphysema, both of which have a high degree of morbidity and significant mortality. Although synthesized primarily by the liver, A1AT has been histochemically localized in monocytes and macrophages in vitro and has been shown to be produced in tissue culture of monocyte-macrophage origin. This study was planned to quantitatively and qualitatively assess the in vivo monocyte-macrophage system contribution to serum A1AT. We used bone marrow transplantation (BMT) as an experimental method because there is commanding evidence that after engraftment, the monocyte-macrophage system of the recipient is replaced by that of donor origin. Protease inhibitor (Pi) typing was done on 150 potential BMT recipients and on their potential donors before transplantation. From these initial recipients, 92 eventually underwent transplantation, and 11 recipient-donor pairs, in which each donor's Pi type contained a band not in the recipient's Pi type, were chosen for the study. Six recipients survived beyond 100 days after BMT, and in these cases the donor contained either an S or an M2 band in his or her Pi type not present in the recipient. Using a silver stain method on diluted serum of known M1M2 and MS types, we were able to detect a 2% dilution of the S band and a 25% dilution of the M2 band. When the same method was applied to gels used in typing recipient Pi after BMT, we were unable to detect any contribution to serum A1AT by the donor monocyte-macrophage system. PMID- 3049863 TI - Anesthesia and evoked potentials: overview. AB - Evoked potentials are increasingly used for intraoperative monitoring. Their use is based on their ability to detect early changes caused by surgical maneuvers which may result in post operative deficits. However, not all changes are surgically related and any decrease in the non surgical causes of evoked potential changes increases the yield of intraoperative monitoring. In this review I will discuss the anesthetic effects on evoked potentials; these include a general description of the anesthetic effects on evoked potentials followed by the effects of premedication, induction, and maintenance agents. Also, described are the effects of adjunct anesthetic agents and techniques. Changes related to anesthesia are not similar and the knowledge of such differences is essential for the planing of anesthesia during the use of evoked potentials. An out line of the anesthetic techniques are described at the end of this review. PMID- 3049865 TI - The founding of Alpha Omega Alpha. PMID- 3049867 TI - Childhood cancer clinical trials and regional tumor registries. PMID- 3049866 TI - Adenocarcinoma in exstrophy and defunctional ureterosigmoidostomy. PMID- 3049868 TI - John Giles Cecil, MD 1855-1913. PMID- 3049869 TI - The use of lasers in the treatment of bladder cancer. PMID- 3049870 TI - Interference. PMID- 3049871 TI - 'Otology at the crossroads' (the second Morell MacKenzie address). AB - It is an honour for an otological surgeon to be asked to deliver the second Sir Morell MacKenzie address. Thank you for the invitation and for that honour. Exactly 100 years ago MacKenzie, with Norris Wolfenden, founded 'The Journal of Laryngology and Rhinology'. 'Otology' was not added to the title until 1892, the year MacKenzie died. I shall return to this theme later. It is also exactly 100 years since, in 1887, MacKenzie was asked by Queen Victoria to examine her son-in law, Frederick, Crown Prince of Germany. The controversy and subsequent recrimination which flowed from the management of Frederick's laryngeal lesion I shall leave to future lecturers to assess. It is as the founder of British laryngology that he should be remembered. As early as 1863 he had started the Metropolitan Free Dispensary for Diseases of the Throat and Loss of Voice, the first institution of its kind in the world. He wrote authoritatively on diseases of the throat and invented many instruments for the difficult art of indirect laryngeal and pharyngeal surgery. He received his Knighthood, also in 1887. PMID- 3049872 TI - Leiomyoma of the nasal cavity. AB - A case of rare leiomyoma of the nasal cavity is reported. The pathological and clinical characteristics of this tumour are discussed. PMID- 3049873 TI - Nasal septal carcinoma: initial symptom of nasal septal perforation. AB - A case of a primary squamous cell carcinoma of the nasal septum in a young female with initial symptom of septal perforation is reported. Carcinoma of the nasal septum is an uncommon entity and there are a few cases reported in the literature. The functional impact of their treatment and the high mortality makes it important to diagnose it at early stage. We discuss the differential diagnosis of septal perforation and recommend early wide surgical excision. PMID- 3049874 TI - Liposarcoma of the larynx. AB - Liposarcomas of the larynx are very rare. Only nine cases appear to have been recorded in the English literature. An additional case of liposarcoma of the larynx occurring in a young patient is reported and the relevant literature discussed. PMID- 3049875 TI - Direct evidence for a stem cell common to hematopoiesis and its in vitro microenvironment: studies on syngeneic (inbred) Wistar Furth rats. AB - When injected into a group of lethally irradiated syngeneic (inbred) Wistar Furth (WF) rats, suspensions of stromal cells grown in monolayer culture from the marrow of WF rats produced hemopoietic colonies in the spleen and rescued 50% of the rats, while 90% of the non-injected (control) rats died within 30 days and had no hemopoietic colonies in the spleens. Fifty percent of the injected (test) rats which died between days 6 and 22 showed hemopoietic regeneration in the bone marrow, while little or no evidence of hemopoietic regeneration was seen in the control animals. Our results suggest that the marrow stroma grown in vitro contain cells with hemopoietic potential and are transplantable. PMID- 3049876 TI - Pentoxifylline (Trental)--a new drug for the treatment of peripheral chronic occlusive arterial disease. AB - Pentoxifylline (Trental), a xanthine analog, was evaluated for tolerance, safety and efficacy in the treatment of chronic arterial disease in a pilot study. Evaluation was performed in 35 cases. Twenty patients (Fontaine stage II or stage III severity) were given pentoxifylline in a daily dose of 1200 mg (Trental 400 t.i.d.) and 15 patients were given placebo for a period of eight weeks, respectively. Pentoxifylline was significantly more effective than the placebo in increasing both the initial and absolute claudication distance (ICD and ACD) in patients with peripheral chronic occlusive arterial disease (COAD). The subjective parameters such as paresthesias, muscular cramps and sensation of heaviness in the legs paralled the course of walking parameters. These results support the hypothesis that pentoxifylline in doses of 400 mg (slow release tablets) t.i.d. enhances blood flow via reducing blood viscosity and improving red cell flexibility in patients with intermittent claudication due to COAD. Pentoxifylline is thus regarded as a promising drug for the treatment of circulatory ischemic disorders, especially in intermittent claudication. It was well tolerated with minimal untoward effects. PMID- 3049878 TI - [Behcet's disease, a current disease]. AB - The treatment of Behcet's is essentially symptomatic and depends on the severity of the manifestations. Colchicine is useful in the minor forms, especially the mucocutaneous, forms. It can be safely used at a dose of 1 mg/day as maintenance treatment to prevent or limit the acute episodes. Other immunomodulators have been proposed : levamisole, disulon, thalidomide, the indications for which are limited by the side effects. Non-steroidal anti-inflammatory agents are indicated in the articular forms. Steroid therapy remains essential in the severe neurological and ophthalmological forms at an initial dose of 1 mg/kg/day. It can be preceded by the intravenous bolus administration of high doses of methylprednisone. Immunosuppressants are useful in the same indications. Although some authors propose them right from the start, we prefer to reserve them for second line treatment in the event of steroid dependence or recurrence, because of their short-term infectious and long-term oncogenic risks. Cyclosporin, which has been proven to be effective by randomised studies, is difficult to manage and responsible for complications, in particular renal. It can therefore only be used as second line treatment by experienced users. Plasmapheresis is reserved as an emergency procedure for functionally threatening forms. Venous or arterial thromboses justify longterm anticoagulation, possibly associated which platelet anti-aggregants. In this chronic disease, the quality of follow-up and the patient's cooperation are essential in order to intervene rapidly and to detect as early as possible the complications of the disease and of the treatment. PMID- 3049877 TI - [Effect of coenzyme A (COA) on the 6-keto-prostaglandin F1 alpha content of the aorta of rats made diabetic with streptozotocin. Potential mechanism of action of its favorable effect on microcirculatory disorders in man]. AB - The production of 6-keto-prostaglandin F1 alpha, a stable metabolite of the vasodilator prostacyclin (PGI2), by fragments of the thoracic aorta of rats made diabetic by streptozotocin, was evaluated by radio-immunological assay. The level of production was found to be statistically higher (p less than or equal to 0.01) in the group treated with Coenzyme A (CoA) than in the placebo group. The authors suggest that the mode of action of CoA 1000 much greater than in microcirculatory pathology could involve an increased production of vasodilator prostacyclin. PMID- 3049879 TI - [Articular manifestations of Behcet's disease. Apropos of 73 cases]. AB - The authors report 73 cases of Behcet's disease with articular manifestations. Polyarthritis, generally considered to be rare, was found in 20.5 per cent of cases in this series. The unusual features are illustrated. Amongst them, two cases presented deforming and destructive lesions, which have been only exceptionally reported in the literature. A third case is unusual because of the association of Gougerot-Sjogren's syndrome with Behcet's disease and polyarthritis deformans. No cases of ankylosing spondylitis were observed. Lastly, the data of this series are compared with those of the literature. PMID- 3049880 TI - [Neurological manifestations of Behcet's disease]. AB - Characterised classically by the association of buccal and genital ulceration and uveitis with hypopyon, Behcet's disease has many other manifestations, amongst which the neurological ones (often referred to as Neuro-Behcet) are important in view of their frequency and their gravity. Anatomically, it produces a subacute haemorrhagic and necrotising meningo-encephalitis, which most typically effects the hypothalamus and brainstem. Clinically, there is extreme polymorphism, central manifestations being the most frequent: seizures, organic brain syndromes, disorders of consciousness, aphasia, hemiplegia, cranial nerve palsies, pseudobulbar and extrapyramidal syndromes and meningism. The peripheral nerves and muscles are rarely affected. Alongside Neuro-Behcet per se, attention has recently been directed to the various cerebro-vascular manifestations, dominated by venous thrombosis. A review of the principal neurological manifestations is given, with comment on anatomico-pathological aspects, clinical presentation, investigational techniques, diagnostic difficulties, prognosis, and treatment. PMID- 3049881 TI - [Behcet's syndrome and the digestive tract]. AB - Gastrointestinal involvement represents only 14% of the anatomical sites of Behcet's syndrome in France. The various segments of the gastrointestinal tract may be affected. Episodes of mucosal aphthae are more frequent in the oesophagus than in the stomach or duodenum. Intestinal involvement constitutes the major gastrointestinal localisation. Colonic or ileo-colonic lesions may appear after several years of recurrent aphthosis and present in the form of acute complications (perforation, massive haemorrhage) or by prolonged haemorrhagic diarrhoea with marked deterioration in the general state. The radiological and endoscopic signs are similar to those observed in various forms of severe acute colitis such as haemorrhagic proctocolitis or Crohn's disease. The diagnosis of an intestinal localisation of Behcet's syndrome is based on the richness of the extra-intestinal signs and moreover on the presence of deep colonic ulcerations frequently situated in healthy mucosa, the presence of an adjacent non-specific inflammatory infiltrate affecting all of the colonic wall, lesions of vasculitis and perivasculitis with signs of leukocytoclasis and fibrinoid necrosis. Surgical treatment is frequently necessary. The high incidence of ulcerative recurrences in the anastomoses, in which fistulae may also develop, requires extensive intestinal resections or diversions by long-term ileostomies. PMID- 3049883 TI - [Arterial involvement in Behcet's disease]. AB - Ten cases of Behcet's disease with arterial lesions were observed in a series of 500 patients over a period of 12 years. The majority of patients were male (9/10) aged between 24 and 36 years with a mean of 30 +/- 5 years. The first group (3 cases) presented with thrombosis of the radial and superficial femoral arteries, the second group (4 cases) presented with aneurysm of the subclavian artery, common and external iliac arteries, brachiocephalic trunk and abdominal aorta and the third group (3 cases) had a combination of thrombosis and aneurysm of the pulmonary, external iliac and renal arteries. PMID- 3049882 TI - [Venous thrombosis in Behcet's disease]. AB - Venous lesions in Behcet's disease (BD) were defined by Adamantiades and represent one of the most suggestive signs of the disease. They are occasionally the first sign of the disease and are frequently the basis for the diagnosis in a case of recurrent thrombosis in a young subject, the preferential context of BD. Involvement of superficial vessels is virtually constant. Venous vasculitis is responsible for non-specific hypersensitivity and erythema nodosa, which constitute some of the major diagnostic criteria. Ocular periphlebitis is one of the elements responsible for posterior uveitis. The originality of the venous involvement is due to the involvement of deep territories. Any vein may be affected, but the remarkable features are the size of the thrombosed vessels: superior and inferior vena cava, iliofemoral veins and the unusual site of the involvement: supra-hepatic veins, cerebral vessels, etc. Inferior vena cava thrombosis may be associated with aneurysms of the pulmonary arteries in the context of Hughes-Stovin syndrome. Cerebral phlebitis, which can now be identified more easily by means of digital angiography, is responsible for a typical picture: headaches, bilateral papilloedema and raised CSF pressure. The classical pictures of optic chiasmatic arachnoiditis and so-called benign intracranial hypertension actually correspond to unrecognised phlebitis. They may also be associated with other neurological lesions. In one half of cases, phlebitis cutaneous manifestations. However, they may precede the diagnostic signs or may occur very late in the course of the disease. They are recurrent and affect a number of different territories.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3049884 TI - [Pathogenic concepts, nosological limits and diagnostic criteria of Behcet's disease]. AB - Behcet's disease, a multi-system disease, raises aetiological and classification problems. Its aetiology is unknown, although several factors have been incriminated in its pathogenesis: immunogenetic, toxic, viral, hormonal,... It overlaps with several groups of diseases: seronegative spondyloarthritis, connective tissue diseases, angiitis, uveomeningitis and aphthosis. Several diagnostic criteria proposed by various authors are discussed. PMID- 3049885 TI - [Medical treatment of Behcet's disease]. AB - The treatment of Behcet's disease is essentially symptomatic and depends on the severity of the manifestations. Colchicine is usefull in the minor, particularly mucocutaneous forms. It can be safely used at a dose of 1 mg/day as maintenance treatment to prevent or limit the acute episodes. Other immunomodulators have been proposed: levamisole, disulon, thalidomide, the indications for which are limited by the side effects. Non-steroidal anti-inflammatory agents are indicated in the articular forms. Steroid therapy remains essential in the severe neurological and ophthalmological forms at an initial dose of 1 mg/kg/day. It can be preceded by the intravenous bolus administration of high doses of methylprednisolone. Immunosuppressants are useful in the same indications. Although some authors propose them right from the start, we prefer to reserve them for second line treatment in the event of steroid dependence or recurrence, because of their short-term infectious and long-term oncogenic risks. Cyclosporin, which has been proven to be effective by randomized studies, is difficult to manage and responsible for complications, in particular renal. It can therefore only be used as second line treatment by experienced users. Plasmapheresis is reserved as an emergency procedure for functionally threatening forms. Venous or arterial thromboses justify long-term anticoagulation, possibly associated with platelet anti-aggregants. In this chronic disease, the quality of follow-up and the patient's cooperation are essential in order to intervene rapidly and to detect as early as possible the complications of the disease and of the treatment. PMID- 3049886 TI - [Non-invasive methods for vascular studies in the diagnosis of deep venous thrombosis]. AB - The sonographic diagnosis of deep venous thrombosis must be made up of a functional continuous wave Doppler study of the whole deep venous system of the limbs, including leg veins, as well as saphenous veins. Then, high resolution B mode real time sonography is used for the detection of direct (echogenic thrombus) or indirect (incompressible vein) signs of thrombosis. This noninvasive approach offers a good sensitivity (about 96%) and a high level of specificity (about 98%). Moreover, B-mode sonography can ensure the differential diagnosis (hematoma, extrinsic compression...) in most cases. So, X-Ray venography is required only when an interventional therapy is planned (thrombectomy, fibrinolysis, inferior vena cava interruption...), or when the noninvasive techniques are not able to show the upper limit of the thrombosis (especially for iliac veins or inferior vena cava), or when there is still a doubt about deep venous thrombosis. Therefore, the number of X Ray venographies can be consistently reduced, thus decreasing both cost and risks. PMID- 3049887 TI - [Evaluation of the diagnostic reliability of combination plethysmography-Doppler. Evaluated on the basis of a prospective study of 176 patients with suspected thrombophlebitis of the limbs including 60 phlebographic correlations]. PMID- 3049888 TI - Biomechanical analysis by chiropractic radiography: Part I. A simple method for determining X-ray projectional distortion. AB - Projectional distortion is known to produce artifactual disrelationships between the images of osseous segments on X-ray film. Apparatus and methods were developed to measure the effect of projectional distortion on a human third lumbar vertebra. The procedures for mathematical analysis of such distortion are outlined, and the results of studies to determine the accuracy of the procedure are presented. PMID- 3049889 TI - Chiropractic in the yellow pages: a content analysis study. AB - This paper presents a content analysis of advertisements by chiropractors in the yellow pages. Information was gathered from 13 cities in the United States for the years 1985 and 1986. Results were categorized by size, number and content of the ads. Estimated expenditures for each city were calculated and a total cost for advertising for the entire chiropractic profession was estimated. Of the 5456 chiropractors listed in the yellow pages in this study, 14.7% bought additional space in the regular listing section, and 11.6% purchased large display advertisements. The remaining 73.7% listed only their name and telephone number. Of those who bought additional space, 10.8% advertised techniques, 11.6% symptoms, 14.7% injuries, 3% professional affiliations and 4% advertised free services. The average annual expenditure for the chiropractor was $2474.00. Future research needs to address the attitudes of the profession and patients toward advertising, and the cost-effectiveness of advertising. The value of the distribution of resources for advertising by chiropractors is questioned. PMID- 3049890 TI - Clinical assessment and treatment of leg length inequalities. AB - Controversy persists regarding the significance of leg length inequality, the diagnostic approach to the use of heel lifts, and the implementation of proper orthopedic support in treatment of anatomical leg length inequalities. The purpose of this paper is to review the literature and formulate a sequential examination for a patient with leg length discrepancy, then outline a formula for the treatment of the patient. The initial problem a practitioner faces in examining a patient with leg length inequality is to determine if a true (anatomical) leg deficiency exists. Through a series of measurements and pelvic assessments, an effective screening process can be accomplished before radiography (scanogram) is required. An understanding of the mechanics along the kinetic chain, anywhere from the foot to the lumbar spine, coupled with the information gained from radiographs, can provide a treatment plan detailing the size of the lift and location. PMID- 3049891 TI - Synovial plicae syndrome. AB - The suprapatellar, medial patellar, and infrapatellar plicae are three embryonic remnants of synovial tissues that are often a cause of intermittent anterior knee pain. A primary blunt trauma or internal pathological derangement will result in the plicae becoming thickened and inelastic with an often inappropriate diagnosis of meniscal tear. The signs, symptoms and examination findings when combined with special testing enables the doctor to correctly establish a diagnosis and proceed with treatment. The chiropractic doctor may play a prime role in the conservative method of treatment and prevent further dysfunction leading to surgical excision as the treatment of choice. This paper will review and summarize the current knowledge of plicae anatomy, pathophysiology, diagnosis and therapeutics in an attempt to provide chiropractors with an increasing awareness of the synovial plicae syndrome. PMID- 3049894 TI - Which philosophy of chiropractic? PMID- 3049893 TI - Conservative management of herniated nucleus pulposes: treatment approaches. AB - Herniated nucleus pulposus (HNP) is a significant cause of physical impairment to the afflicted individual and a major factor in a wide range of socioeconomic issues. This paper examines various disc herniation therapies and compares conservative treatment modes with surgical intervention. Results of a 10-yr study are reviewed and recommendations are made concerning conservative treatment management in the physician's office and at home, return-to-work considerations, preventive care and the importance of cooperation among patient, physician, employer and third-party provider. The authors conclude that conservative treatment is an acceptable, cost-effect alternative to surgery. PMID- 3049892 TI - Conservative care of urinary incontinence in the elderly. AB - Urinary incontinence is a common, costly and demoralizing problem among the elderly. Remedial efforts are often not attempted owing to the misconception that incontinence is an inevitable and irreversible characteristic of aging. In fact, a variety of relatively conservative methods of reducing geriatric incontinence are available. This paper reviews the categories of incontinence, outlines assessment strategies and critiques the literature on biofeedback, exercise, behavior therapy and electrical stimulation as treatments for geriatric incontinence, and briefly considers a role for the chiropractic physician. PMID- 3049895 TI - Unity--facing the RVS. PMID- 3049896 TI - The U.S. constitution in perspective. PMID- 3049897 TI - Computer retardation. PMID- 3049898 TI - GaIN: a network of physicians and hospitals in Georgia. PMID- 3049899 TI - The National Library of Medicine, computers, and the garbage can method of problem solving. PMID- 3049900 TI - Surgical unit time utilization review: resource utilization and management implications. AB - As health care providers seek ways to reduce the cost of health care services, hospital operating rooms (ORs) have been identified as potential areas for cost reduction efforts. Cost containment efforts which have shifted significant portions of the inpatient population to ambulatory areas have resulted in an inpatient population which is sicker and more procedure-intensive. Efficient management of operating rooms has assumed even greater importance in this environment. Inefficient or inaccurate scheduling of OR time often results in delays of surgery or cancellations of procedures, which are costly to the patient and the hospital. Approaches to efficient use of ORs include computerized scheduling, utilization monitoring, and refinement of scheduling policies and procedures. In the absence of commercially available software to meet operating room management information needs, Johns Hopkins developed its own system in 1983. This software provides detailed information for daily OR management and long-term planning. The computerized operating room scheduling and monitoring system is described in this article and an operational measure of scheduling accuracy is proposed. Suggestions are made for incorporating this measure into planning and allocation decisions. PMID- 3049901 TI - The interpretation of pituitary gonadotrophin assays--a continuing challenge. PMID- 3049902 TI - Suppression of LH response to gonadotrophin-releasing hormone or oestradiol by ACTH(1-24) treatment in anoestrous ewes. AB - Stress is known to result in lowered female reproductive efficiency. The objective of this study was to examine how increased pituitary-adrenal activity may influence gonadotrophin release in anoestrous ewes. Various doses (0.06-1.0 mg) of a synthetic adrenocorticotrophic hormone (ACTH(1-24)) preparation were injected into ewes 30 min or 3 h before an i.v. injection of 500 ng gonadotrophin releasing hormone (GnRH). The LH response to GnRH given 30 min after ACTH(1-24) was similar to that after GnRH alone, whereas the response 3 h after ACTH(1-24) was significantly lower, irrespective of the dose of ACTH(1-24). At 30 min and 3 h after ACTH(1-24) the concentrations of cortisol exceeded 50 nmol/l compared with baseline values of less than 10 nmol/l. The effect of ACTH(1-24) on oestradiol-induced LH release was also examined. Those ewes receiving 0.8 mg ACTH(1-24) depot and 50 micrograms oestradiol benzoate simultaneously had a preovulatory-type increase in LH 14-20 h later, similar to when oestradiol benzoate was given alone. None of the ewes receiving an additional 0.8 mg ACTH(1 24) depot 10 h after oestradiol benzoate had increases in LH concentration. The cortisol concentrations in all ewes receiving either one or two injections of ACTH(1-24) were greater than 35 nmol/l at 10 h after the oestradiol injection. However, concentrations of progesterone increased from 0.9 +/- 0.3 (S.E.M.) nmol/l at the time of the second ACTH(1-24) injection to 2.1 +/- 0.3 nmol/l after 2 h.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3049903 TI - Effect of glucose concentrations on functional maturation of monolayer-cultured B cells from the rat pancreas. AB - The effect of two concentrations of glucose on the maturation of the response of B cells was studied in pancreatic monolayer cultures of the neonatal rat using a perifusion system. After exposure for the initial 3 days to a medium with 5.5 mmol glucose/l and 10 mumol iodoacetic acid/l (day 3), in order to delete fibroblasts selectively, monolayer cultures were kept for a total of 12 days in medium with either 5.5 or 16.7 mmol glucose/l alone. On day 3, B cells responded in a monophasic fashion, with no significant second phase, to acute challenge with 16.7 mmol glucose/l. At a concentration of 10 mmol/l, leucine and 2 ketoisocaproate both produced only minimal increases in the second phase of secretion above the basal level. In contrast, B cells on day 7 cultured in 5.5 mmol glucose/l showed a biphasic response to glucose, leucine and 2 ketoisocaproate. The magnitude in response to glucose was well preserved at day 15 of culture, whereas the stimulatory effects of leucine and 2-ketoisocaproate decreased to 24-57% of that of B cells on day 7. Moreover, B cells on day 7 cultured in 16.7 mmol glucose/l responded in a biphasic manner to glucose, the response being 65% of that of B cells in 5.5 mmol glucose/l. Additionally, the response to leucine and 2-ketoisocaproate still appeared to be monophasic. At day 15 of culture, however, the response of B cells in 16.7 mmol glucose/l to glucose was 105%, to leucine 245% and to 2-ketoisocaproate 127% of that of B cells in 5.5 mmol glucose/l.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3049904 TI - The human neutrophil serine proteinases, elastase and cathepsin G, can mediate glomerular injury in vivo. AB - We infused microgram quantities of active or inactive PMN elastase and cathepsin G into the renal arteries of rats. Both active and inactive elastase localized to the glomerular capillary wall equally, and in amounts that could be achieved physiologically in GN. However, elastase-perfused rats developed marked proteinuria (196 +/- 32 mg/24 h) compared with control rats receiving inactive elastase (19 +/- 2 mg/24 h, p less than 0.005). Similar results were seen with active and inactive cathepsin G. Neither elastase nor cathepsin G infusion was associated with histologic evidence of glomerular injury. We conclude that the PMN neutral serine proteinases elastase and cathepsin G can mediate marked changes in glomerular permeability in vivo due to their proteolytic activity, and thus, may contribute to the proteinuria observed in PMN-dependent models of GN. PMID- 3049905 TI - Isolation and immunocytochemical characterization of human bone marrow stromal macrophages in hemopoietic clusters. AB - Stromal macrophages (M phi) have been localized in situ and isolated within erythroid clusters from human marrow. Stromal M phi arborize in an extensive network uniformly distributed throughout marrow interstitium, and express the phenotype CD4+, CD11a+, CD11c+, CD13+, CD14+, CD16+, CD18+, CD31+, CD32+, FcRI+, HLA-DR+, and CD35-, transferrin receptor-negative, and CD11b (weak). They express endocytic receptor antigens, but show significant differences in myeloid antigen expression compared with freshly harvested or cultured monocytes. Human stromal M phi are therefore specialized mature marrow M phi that are accessible for further investigations in infectious, storage, or hemopoietic disorders. PMID- 3049906 TI - Regulation of murine MHC class II molecule expression. Identification of A beta residues responsible for allele-specific cell surface expression. AB - A panel of mutant class II genes have been constructed using site-directed mutagenesis and DNA-mediated gene transfer. Using this technique, Ak beta polypeptides have been altered by substituting one or more Ad beta-specific residues at polymorphic positions in the beta 1 domain. Transfection of M12.C3 B lymphoma cells with most mutant Ak beta* genes results in the expression of Ak beta* Ad alpha molecules on the cell surface. However, the substitution of a single d allele residue at position 78 or 86 in the Ak beta polypeptide results in either the complete absence or very low levels, respectively, of cell surface expression of the Ak beta* Ad alpha molecule, but does not alter Ak beta* Ak alpha expression. The T.86 Ak beta* Ad alpha is expressed primarily in an intracellular compartment while the T.78 Ak beta* molecule does not appear to be produced. The core-glycosylated T.78 Ak beta* polypeptide does, however, form a complex intracellularly with the core-glycosylated Ii polypeptide. Substitution of the combination of d allele residues at Ak beta polymorphic positions 9, 12, 13, 14, and 17 results in the absence of Ak beta* Ak alpha cell surface expression but does not alter the expression of this mutant Ak beta* polypeptide with the Ad alpha polypeptide. These allele-specific expression mutants demonstrate that substitution at certain beta 1 domain positions may result in the alteration of Ia cell surface expression and that the transport of Ia molecules from the Golgi apparatus to the cell surface may be regulated by signals that are determined by the interaction of polymorphic residues in both the alpha and beta polypeptides. PMID- 3049907 TI - Interleukin 4 causes isotype switching to IgE in T cell-stimulated clonal B cell cultures. AB - Although it has been established that IL-4 enhances both IgG1 and IgE secretion in LPS-stimulated B cell cultures, these studies failed to determine whether IL-4 preferentially induces isotype switching or preferentially allows for the maturation of precommitted precursor cells. To distinguish between these possibilities, it is necessary to ascertain the effect of IL-4 on the isotypes secreted by individual precursor cells during clonal expansion. Therefore, clonal cultures of B cells stimulated with a Th2 helper cell line specific for rabbit Ig and rabbit anti-mouse IgM were established. The majority of B cells are capable of undergoing clonal expansion under these conditions. To vary the level of IL-4 present, either IL-4 or anti-IL-4 was added to cultures. In the presence of IL-4 there was an increase in the proportion of clones that secreted IgE and a decrease in the proportion of clones that secreted IgM. The addition of IL-4 to cultures also increased the amount of IgE secreted by individual clones. Thus, these experiments definitively prove that IL-4 causes specific heavy chain class switching to IgE in Th2-stimulated B cell cultures. In contrast, IL-4 does not affect the proportion of clones secreting IgG1, suggesting that other consequences of Th cell-B cell interactions play a role in the generation of an IgG1 response. PMID- 3049909 TI - Human monoclonal antibodies to group B streptococcus. Reactivity and in vivo protection against multiple serotypes. AB - Group B streptococcal (GBS) infections cause significant mortality and morbidity among infants. Passive antibody immunotherapy has been proposed as treatment for infected infants. To this end, two human mAb-secreting cell lines were produced by EBV immortalization of human B cells. The mAbs were specific for the group B polysaccharide and bound to strains of all five serotypes as demonstrated by ELISA and crossed immunoelectrophoresis. The mAbs reacted and opsonized 100% (132/132) of the clinical isolates tested which represented all four capsule types. Both prophylactic and therapeutic protection with these mAbs were demonstrated in neonatal rats given lethal infections of types Ia and III human clinical isolates. These data indicate that a single human mAb directed against the group B carbohydrate can protect against GBS infections caused by the different serotypes. This antibody may be useful in the passive immunotherapy of infants infected with GBS. PMID- 3049908 TI - Synergism of BSF-2/interleukin 6 and interleukin 3 on development of multipotential hemopoietic progenitors in serum-free culture. AB - We investigated the effects of B cell stimulatory factor 2/interleukin 6 (BSF 2/IL-6) on the development of murine hemopoietic progenitors using serum containing culture and serum-free culture. In serum-containing culture, BSF-2 mainly supported multipotential blast cell colonies from spleen cells of normal and 5-fluorouracil (5-FU)-treated mice. In serum-free culture, no colony growth was seen in the presence of BSF-2. Addition of BSF-2 to the serum-free culture containing IL-3 resulted in a significant increase in the number of colonies formed from multipotential progenitors in spleen cells and bone marrow cells of 5 FU-treated mice, whereas no effects were seen on the number of single or oligolineage colonies formed by the spleen cells of normal mice. These results suggested that BSF-2 and IL-3 act synergistically on the multipotential progenitors but not on the maturer progenitors. When BSF-2 was added to a culture containing low concentrations of IL-3 (1 U/ml, 4 U/ml), which had little effect on colony formation, the number of total colonies formed by the spleen cells and bone marrow cells of 5-FU-treated mice increased significantly. The combination of BSF-2 and 40 U/ml of IL-3 resulted in a significant enlargement of GMM colonies. Thus, BSF-2 appears to enhance the sensitivity of multipotential hemopoietic progenitors to IL-3. PMID- 3049911 TI - Cytoadherence by Plasmodium falciparum-infected erythrocytes is correlated with the expression of a family of variable proteins on infected erythrocytes. AB - Plasmodium falciparum-infected erythrocytes (IRBCs) adhere specifically to venular endothelium and thereby evade spleen-dependent immune mechanisms. We have investigated the molecular basis of cytoadherence. We report here that the capacity for cytoadherence of IRBCs is correlated with the expression of a family of variable proteins on the surface of IRBCs. Essential to these studies was the use of in vitro techniques for modulating the cytoadherence phenotype of cloned parasites. In initial studies, we found culture-adapted parasites to be poorly cytoadherent or noncytoadherent. To select for cytoadherent parasites, we incubated knobbed IRBCs with C32 melanoma cells and cultured the adherent cells. Repeated rounds of selection produced parasites with increased cytoadherence. To select for noncytoadherent parasites, we cultured the cells that did not adhere to C32 melanoma cells. Cytoadherent IRBCs from two different cloned isolates had large (Mr greater than 2.4 x 10(5) radioiodinatable proteins that differed in size between the isolates but had in common the biochemical properties of trypsin sensitivity and insolubility with Triton X-100. The proteins were not detected with uninfected erythrocytes, indicating that they were parasite determined, nor were they detected with IRBCs containing parasites cultured for many months without selection. With continued selection for the cytoadherent phenotype, additional IRBC surface proteins with larger molecular sizes (Mr 2.9 x 10(5) and 3.2 x 10(5] appeared. A sequence of reversible changes in the cytoadherence phenotype of cloned parasites was accompanied by variation in the molecular size of the IRBC surface protein. Increased cytoadherence was correlated with expression of larger proteins and decreased cytoadherence was correlated with expression of smaller proteins; there was no change in the molecular size of two other parasite proteins associated with the IRBC membrane. The results indicate that the expression of this family of proteins is closely linked to the cytoadherence phenotype of the parasites, suggesting that the members of the protein family have a role in mediating cytoadherence between IRBCs and endothelial cells. PMID- 3049910 TI - Synthesis and release of platelet-activating factor is inhibited by plasma alpha 1-proteinase inhibitor or alpha 1-antichymotrypsin and is stimulated by proteinases. AB - TNF and IL-1 stimulate the synthesis and release of platelet-activating factor (PAF) by neutrophils and vascular endothelial cells. Serum inhibits PAF production even after inactivation of an acetylhydrolase that degrades PAF. Human plasma was fractionated by gel filtration chromatography, and two inhibitory fractions were detected, one containing PAF-acetylhydrolase activity and the other alpha 1-proteinase inhibitor. Low concentrations of this antiproteinase and of human plasma alpha 1-antichymotrypsin inhibited TNF-induced PAF synthesis in neutrophils, macrophages, and vascular endothelial cells. Both antiproteinases also inhibited PAF production stimulated by phagocytosis in macrophages and induced with IL-1 in neutrophils or with TNF in vascular endothelial cells. These results suggest that a proteinase activated on the plasma membrane or secreted by these cells is involved in promoting PAF synthesis. Indeed, addition of elastase to macrophages, neutrophils, and endothelial cells stimulated synthesis and release of PAF much faster than TNF. A similar stimulation was observed in incubations with cathepsin G. To identify a proteinase activated in TNF-treated cells, neutrophils and endothelial cells were incubated with specific chloromethyl ketone inhibitors of elastase and cathepsin G. Synthesis of PAF was significantly inhibited by low concentrations of the cathepsin G inhibitor. The finding that antiproteinases are inhibitory at concentrations 100-fold lower than those present in plasma raises questions as to the ability of TNF and IL-1 to stimulate neutrophils in circulation or endothelial cells to synthesize PAF. We propose that PAF production is limited to zones of close contact between cells, which exclude antiproteinases. PMID- 3049913 TI - Prevention of experimental cerebral malaria by anticytokine antibodies. Interleukin 3 and granulocyte macrophage colony-stimulating factor are intermediates in increased tumor necrosis factor production and macrophage accumulation. AB - IL-3 and granulocyte/macrophage colony stimulating factor (GM-CSF) are two cytokines released by activated T lymphocytes that stimulate the growth and differentiation of various hematopoietic cell lines, among which are macrophages. It has been shown that TNF/cachectin, another cytokine that is released mostly by activated macrophages, plays a central role in experimental cerebral malaria (CM), an acute and lethal neurological syndrome induced by Plasmodium berghei ANKA infection in CBA mice. Since CM requires functional CD4+ T lymphocytes to occur, we explored, by injecting rabbit antibodies to murine rIL-3 and/or GM-CSF, whether these cytokines are intermediates in the marked TNF release leading to CM. Treatment of infected mice with each antibody separately had no protective effect. In contrast, when both anti-rGM-CSF and anti-rIL-3 antibodies were injected together; (a) the occurrence of neurological syndrome was prevented in 90% of the cases; (b) the rise in serum TNF was prevented; and (c) macrophage accumulation in the spleen was significantly reduced. Murine CM appears to involve a cytokine cascade in which IL-3 and GM-CSF lead to the accumulation of TNF-releasing macrophages in vivo. PMID- 3049914 TI - Unexplained lymphadenopathy in family practice. An evaluation of the probability of malignant causes and the effectiveness of physicians' workup. AB - This study reported here was undertaken to determine the probability of malignancy in patients presenting with unexplained lymphadenopathy in primary care practice and to estimate the effectiveness of current referral patterns by family physicians in relation to malignant disease. Clinical characteristics that may be discriminatory for malignant causes were also investigated. A retrospective analysis was performed of 82 patients who underwent biopsy for unexplained lymphadenopathy from 1982 to 1984; data regarding the incidence of unexplained lymphadenopathy and the referral rate for this problem were obtained from registration projects. A total of 29 malignant lymphadenopathies were identified for a prior probability of 1.1 percent and a posterior (after referral) probability of 11 percent. The ability of the family physician to refer malignant cases within four weeks after initial consultation (sensitivity of referral) was 80 to 90 percent; 91 to 98 percent of benign cases were not referred (specificity of referral). An increased likelihood of malignancy was associated with age over 40 years (4 percent) and supraclavicular lymphadenopathy (50 percent). The incidence of malignancy in patients presenting with unexplained lymphadenopathy to the family physicians is very low (1 to 2 percent). Nevertheless, despite the paucity of validated discriminatory factors, the family physicians perform a reasonably effective selection process toward referral and biopsy. PMID- 3049915 TI - The comprehensiveness of computer-assisted searches of the medical literature. AB - Bibliographic searches using MEDLINE, the National Library of Medicine computerized database, can usually be done in less time and with greater specificity than searches using Index Medicus. To use a computer-assisted bibliographic retrieval system to full advantage, it is necessary to understand the indexing system. The study reported here compares both the number and the relevance of references retrieved using various search terms for two clinical questions. Based on the outcomes of these searches, recommendations are made for clinicians who plan to use MEDLINE services. PMID- 3049912 TI - A novel activation pathway for mature thymocytes. Costimulation of CD2 (T,p50) and CD28 (T,p44) induces autocrine interleukin 2/interleukin 2 receptor-mediated cell proliferation. AB - Prior studies have shown that thymocytes, unlike peripheral T cells, do not proliferate in response to mitogenic combinations of anti-CD2 mAbs. The present study demonstrated that stimulation by a mitogenic anti-CD2 combination (9-1 plus 9.6) with anti-CD28 induced vigorous thymocyte proliferation in the absence of exogenous IL-2. This thymocyte proliferation was IL-2 dependent as shown by the complete inhibition using anti-IL-2-R mAbs. Induction of IL-2-R transcripts was detected in thymocytes stimulated by the anti-CD2 antibody combination alone or the anti-CD2 combination plus anti-CD28 antibody. However, induction of IL-2 transcripts was observed only in thymocytes triggered jointly by the anti-CD2 combination plus anti-CD28 antibodies. The double-negative (CD4-8-) or CD1+ thymocytes isolated by sorting or by panning were unresponsive to CD2/CD28 triggering. The same mitogenic signal could induce vigorous proliferation of thymocytes with a mature phenotype, i.e., CD3+CD4+ or CD3+CD8+ thymocytes. Immunofluorescence studies demonstrated that the majority of CD3+ thymocytes were CD28+, and most of the CD28+ cells were located in the medullary compartment of thymus. These results indicated that the T cell lineage surface molecules CD28 and CD2 are involved in the regulation of expansion and further differentiation of mature thymocytes. PMID- 3049916 TI - Aging, infections, and the immune system. AB - There is increasing clinical and laboratory evidence of decline in the immune system in the elderly patient with a simultaneous rise in the incidence of certain infections. Along with the involution of the thymus gland with age, there is evidence of decline in both T- and B-lymphocyte function and also in delayed hypersensitivity. In addition, there is evidence of an increase in various autoantibodies as a person ages. In recent years evidence has been presented of a genetic basis to this declining system. Because of these changes and because severe infections present more subtly in the elderly patient than in the young, the physician's suspicion for serious infections in the elderly should be heightened and immunization programs in the elderly adhered to. PMID- 3049918 TI - A medical model for criminalistics education. AB - The history of medical education during the period of 1870 to 1926 is examined in the context of current issues confronting education in the forensic laboratory sciences. Medical education was radically altered during this period, changing from a rudimentary lecture/apprenticeship system into its modern form. Although the motivating forces had developed over some time, the actual change was quite rapid. By examining how this change occurred, we gain insight into how changes in our own profession might be initiated. Parallels between our current situation and that in medical education 117 years ago include: (1) the primary burden of professional education is borne outside the university in an apprenticeship system, (2) the apprenticeship system is overburdened by a dramatic expansion in the knowledge and skills needed for professional practice, (3) there is no standardized curriculum or accreditation process for educational programs, and (4) there is no educational program that incorporates formal clinical education. Based on this historical analysis, three major goals are proposed: (1) active entreprenurial promotion of professional educational programs by academics, (2) creation of a committee within the American Academy of Forensic Sciences to critique and rate university programs, and (3) the development of a well-defined clinical education program. A model for formalized clinical education in the forensic laboratory sciences is proposed, incorporating clinical professors, student clerkships, and university control over instruction within an operational forensic science laboratory. Benefits from this arrangement include: efficient combination of physical plants, added personnel resources in the laboratory, rapid introduction of research into the laboratory, enhanced prestige for both academics and practitioners, and relief of the laboratory's in-house training burden. PMID- 3049919 TI - Frank C.Coleman, M.D. Man of many talents. PMID- 3049920 TI - Astrology and the PRO. PMID- 3049921 TI - Adnexal torsion: five-year experience at Tampa General Hospital. PMID- 3049917 TI - Specialty bias in obstetric care for high-risk socioeconomic groups in Maine. AB - From 1982 to 1984, 46,501 infants were born in Maine hospitals in 46,286 deliveries, of which 6,343 were born to women on state Medicaid (Title 19), and 6,307 were born to women with no health insurance. In comparison with others born in Maine during those years, more infants in these presumed low socioeconomic groups died, were transferred immediately to other hospitals, had low birthweights, or were readmitted to a hospital within 30 days of birth. Of all deliveries, 105 family physicians or general practitioners performed 22 percent, 82 obstetricians performed 69 percent, and 16 osteopathic physicians performed 5 percent; but of Medicaid deliveries, obstetricians delivered only 59 percent, while family physicians-general practitioners and osteopaths did commensurately more. The decreased proportion of Medicaid patients cared for by obstetricians was especially prominent in Maine's urban hospital service areas. Pediatricians, on the other hand, cared for the same proportion of Medicaid children as they did all children in all hospital service areas in the state. The distribution of low socioeconomic, higher obstetric risk patient groups among various medical specialties as demonstrated in these data should be considered by health planners, malpractice insurers, and health insurers including state Medicaid programs. PMID- 3049922 TI - Daddy, tell me a ghost story. PMID- 3049924 TI - A Victorian mother's vigil over her sick son: Emma Gilpin's journal of 1888. PMID- 3049923 TI - W.C. Chowning, East Coast physician/politician. PMID- 3049925 TI - The medical profession in Florida 1821-1874. PMID- 3049926 TI - William Hughlett and friends: early medicine along the Indian River. PMID- 3049928 TI - History of hysterectomy. PMID- 3049927 TI - The summer nightmare: poliomyelitis in Florida. PMID- 3049930 TI - Successful pregnancy in a renal transplant recipient taking cyclosporine A: report of a case. PMID- 3049929 TI - Medical care system: bigger but not better. PMID- 3049933 TI - A study of the role of the hexose monophosphate pathway with respect to fatty acid biosynthesis in sporulation of Saccharomyces cerevisiae. AB - 13C NMR was used to study the pattern of label incorporation from [2-13C]acetate into trehalose during sporulation in Saccharomyces cerevisiae. A wild-type strain and a strain homozygous for the zwf1 mutation (which affects glucose-6-phosphate dehydrogenase) were used. In the wild-type it was possible to deduce the cycling of glucose 6-phosphate around the hexose monophosphate pathway whilst in the mutant strain this did not occur. The requirement of the hexose monophosphate pathway for providing NADPH for fatty acid biosynthesis was examined using 13C NMR and GC/MS. The wild-type strain produced a typical profile of fatty acids with palmitoleic acid being the most abundant whereas the mutant contained only one-quarter the amount of total fatty acid. As zwf1 homozygous diploids are able to sporulate this indicates that the large amount of fatty acid biosynthesis observed in sporulation of wild-type strains is not essential to the process. PMID- 3049932 TI - Induction of a stable morphological change in Propionibacterium freudenreichii. AB - When cells of Propionibacterium freudenreichii were incubated under fasting conditions and then plated in the presence of an inhibitor of protein synthesis, a variable but significant (greater than 10(-2) fraction of the population changed their morphology from rod to sphere, with a considerable thickening of the cell wall. This change was accompanied by metabolic and antibiotic-resistance modifications, including the synthesis of at least one new enzyme (alpha glucosidase), and by the simultaneous appearance of several new species of DNA, presumably plasmids. The round cells grew faster than the parent strain and maintained their morphology indefinitely when propagated on complex medium containing glucose as the main carbon source. However, when glucose was omitted, cells returned to the rod form and regained their previous characteristics, including the absence of detectable plasmids. PMID- 3049931 TI - Metabolic regulation of the K(ATP) and a maxi-K(V) channel in the insulin secreting RINm5F cell. AB - K channels in the cell membrane of the insulin-secreting RINm5F cell line were studied using the patch-clamp technique in cell-attached patch mode. With 140 mM K in the pipette, two channels displaying different conductive and kinetics properties were observed. A voltage-independent, inward-rectifying, 55-pS channel was active at rest (no glucose, -70 mV), but was almost completely inhibited by 5 mM glucose. A 140-pS channel was seen in the absence of glucose only after cell membrane depolarization with high (30 mM) K. This channel was voltage dependent, with a linear slope conductance between -60 and +60 mV, and was completely inhibited only by greater than 15 mM glucose. The former channel we identify as an ATP-sensitive channel previously described in excised patches and refer to it as the K(ATP) channel. The latter, because of its large conductance and voltage dependent kinetics, will be referred to as the maxi-K(V) channel, adopting a nomenclature previously used to classify highly conductive K channels (Latorre, R., and C. Miller, 1983, Journal of Membrane Biology, 71:11-30). In addition to glucose, mannose and 2-ketoisocaproate, which also initiate insulin secretion and electrical activity in the islet beta cell, reduced the activity of both the K(ATP) and the maxi-K(V) channel. Lactate and arginine, which potentiate but do not initiate insulin secretion or beta cell electrical activity in normal islets, each caused a large reduction in maxi-K(V) channel activity, without consistently affecting the activity of K(ATP) channels. Another agonist that potentiates insulin secretion and electrical activity in normal cells, the tumor-promoting phorbol ester TPA, blocked maxi-K(V) channel activity while stimulating the activity of the K(ATP) channel, thereby implicating phosphorylation in the control of channel activity. These results indicate that metabolic substrates that initiate electrical activity and insulin secretion in normal beta cells reduce the activity of both the K(ATP) and the maxi-K(V) channel, while potentiating agents reduce only the maxi-K(V) channel. The possible role of these two channels in the processes of initiation and potentiation of the beta cell response is discussed. PMID- 3049935 TI - Aspects of the regulation of adenylate cyclase synthesis in Escherichia coli K12. AB - In Escherichia coli K12 expression of the adenylate cyclase gene is subject to multiple controls. In order to gain understanding of the regulation of adenylate cyclase synthesis, operon and protein fusions were constructed by in vitro recombination either into bacteriophage lambda or low-copy-number plasmids, or directly on the chromosome at the cya locus. The fusions were used in physiological experiments as probes to study transcriptional and translational controls of cya expression. It was found that adenylate cyclase synthesis was insensitive to glucose effects. As already described by other workers, the CAP cAMP complex had a moderate negative control on cya expression. In addition it was observed that concomitant with a severe slackening of growth rate, specific to the growth of cya strains in rich medium, cya expression was considerably enhanced. This increase of adenylate cyclase synthesis did not appear to be directly dependent on the presence of a functional cAMP receptor (CAP), and seemed to be controlled at the level of transcription. Finally, translation of the cya message was very weak when compared to cya transcription (the mRNA level was the same in protein and operon fusions. PMID- 3049936 TI - Transfer of the Ti plasmid from Agrobacterium tumefaciens into Escherichia coli cells. AB - We have screened strains of Agrobacterium tumefaciens for spontaneous mutants showing constitutive transfer of the nopaline Ti plasmid pTiC58 during conjugation. The Ti plasmid derivatives obtained could be transferred not only to A. tumefaciens but also to E. coli cells. The Ti plasmid cannot survive as a freely replicating plasmid in E. coli, but it can occasionally integrate into the E. coli chromosome. However, insertion in tandem of plasmids carrying fd replication origins (pfd plasmids) into the T-DNA provides an indicator for all transfer events into E. coli cells, providing fd gene 2 protein is present in these cells. This viral protein causes the excision of one copy of the pfd plasmid and allows its propagation in the host cell. By using this specially designed Ti plasmid, which was also made constitutive in transfer functions, we found plasmid exchange among A. tumefaciens strains and between A. tumefaciens and E. coli cells to be equally efficient. A Ti plasmid with repressed transfer functions was transferred to E. coli with a rate similar to the low frequency at which it was transferred to A. tumefaciens. The expression of transfer functions of plasmid RP4 either in A. tumefaciens or in E. coli did not increase the transfer of the Ti plasmid into E. coli cells, nor did the addition of acetosyringone, an inducer of T-DNA transfer to plant cells. The results show that A. tumefaciens can transfer the Ti plasmid to E. coli with the same efficiency as within its own species. Conjugational transmission of extrachromosomal DNA like the narrow-host-range Ti plasmid may often not only occur among partners allowing propagation of the plasmid, but also on a 'try-all' basis including hosts which do not replicate the transferred DNA. PMID- 3049934 TI - Genetic regulation of the quinic acid utilization (QUT) gene cluster in Aspergillus nidulans. AB - A large number of quinic acid non-utilizing qut mutants of Aspergillus nidulans deficient in the induction of all three quinic acid specific enzymes have been analysed. One class the qutD mutants, are all recessive and are non-inducible at pH 6.5 due to inferred deficiency in a quinate ion permease. Two regulatory genes have been identified. The QUTA gene encodes an activator protein since most qutA mutants are recessive and non-inducible although a few fully dominant mutants have been found. The QUTR gene encodes a repressor protein since recessive mutations are constitutive for all three enzyme activities. Rare dominant non inducible mutants which revert readily to yield a high proportion of constitutive strains are inferred to be qutR mutants defective in binding the inducer. The gene cluster has been mapped in the right arm of chromosome VIII in the order: centromere - greater than 50 map units - ornB - 12 map units - qutC (dehydratase) 0.8 map units-qutD (permease), qutB (dehydrogenase), qutE (dehydroquinase), qutA (activator)-4.4 map units - qutR (repressor)-20 map units - galG. This organization differs from that of the qa gene cluster in Neurospora crassa, particularly in the displacement of qutC and qutR. PMID- 3049937 TI - Inactivation of human alpha-1-antitrypsin by a tissue-destructive protease of Legionella pneumophila. AB - Three extracellular proteases produced by Legionella pneumophila during growth in liquid medium were examined for their effects on human alpha-1-antitrypsin (alpha 1-AT). One of these proteases, tissue-destructive protease (TDP) destroyed completely the trypsin-inhibitory capacity of alpha-1-AT at protease: inhibitor molar ratios down to 0.002:1. After inactivation by TDP, the Mr of alpha-1-AT was reduced by 5000 in SDS-PAGE. This suggested that inactivation entailed only limited cleavage. PMID- 3049938 TI - Attachment of influenza C virus to human erythrocytes. AB - Binding experiments with radioactively labelled influenza C virions were carried out to investigate the interaction of the virus with human erythrocytes. The erythrocytes from any of 35 different individuals were found to contain influenza C virus-binding sites though their number was variable among the individuals and was much less than that on mouse, rat and chicken erythrocytes. Attachment of influenza C virus to human erythrocytes was inhibited completely by prior treatment of the virus with anti-HE monoclonal antibody having a strong haemagglutination inhibition activity. Pretreatment of erythrocytes with neuraminidase or the neuraminate-O-acetylesterase of influenza C virus resulted in a marked reduction in the level of virus binding. Thus it appears that human erythrocytes have a low level of O-acetylated sialic acid-containing glycoconjugates that can interact specifically with the HE glycoprotein of influenza C virus. Proteolytic digestion of erythrocytes with ficin, bromelain or V-8 protease inhibited virus binding almost completely, suggesting that the erythrocyte receptor for influenza C virus is a glycoprotein. In contrast to these enzymes, trypsin treatment of erythrocytes reduced virus binding by only about 50%, and alpha-chymotrypsin treatment did not inhibit at all. It was also found that treatment of erythrocytes with monoclonal antibody to the M or N blood group antigen greatly inhibited virus binding to the cells. These results, taken together, suggest that most influenza C virus receptors on human erythrocytes, if not all, reside on glycophorin A which is known to possess the M or N blood group activity. PMID- 3049939 TI - In vivo expression of rubella antigens on human leucocytes: detection by flow cytometry. AB - Flow cytometry has been used to detect in vivo expression of rubella antigens on human leucocytes. Sequential samples of peripheral blood were obtained from four volunteers with naturally acquired rubella and five persons immunised with RA27/3 rubella vaccine. Leucocytes were stained for rubella antigens using a pool of rubella monoclonal antibodies. Rubella antigens were detected on the leucocytes of all four volunteers with naturally acquired rubella between 1 and 13 days after onset of illness. Viral antigens were expressed more frequently on the monocyte (9-51%) than the lymphocyte (less than 1-4%) and granulocyte (less than 1-3%) populations. Among the vaccines, rubella antigens were detected on the leucocytes of four of the five volunteers between 5 and 12 days after immunisation. The expression of viral antigens was more transient and the proportion of cells exhibiting rubella-specific fluorescence considerably lower following vaccination (1-12%) than natural infection (9-51%). Our results demonstrate that flow cytometry provides a rapid and sensitive analytical technique for detecting viral antigens on leucocytes from infected persons. Leucocytes may play an important role in the pathogenesis of rubella infection. PMID- 3049941 TI - Electronic information for physicians: a new dimension in solving traditional problems. PMID- 3049942 TI - Altered expression of the D1.1 ganglioside in the cerebellum of the Weaver mouse. AB - Expression of the D1.1 ganglioside was studied immunohistochemically in developing cerebella from normal and weaver mutant mice. In the normal cerebellum at postnatal day 7 (P7), D1.1 expression was restricted to the external granule cell layer (EGL). At later ages, D1.1 disappeared as the developing granule neurons ceased mitosis and began migrating toward the internal granule-cell layer. In the weaver cerebellum, D1.1 was expressed in the EGL in apparently normal fashion at P7, but failed to disappear at later ages. As late as P35, D1.1 immunoreactivity was observed throughout the weaver cerebellar cortex. The relative amounts of D1.1 ganglioside in weaver and normal cerebella were compared by thin layer chromatography of total gangliosides, followed by overlay of the chromatogram with anti-D1.1 and 125I-labelled second antibody. Autoradiograms showed that at P12 and P35 the weaver tissue contains six- to tenfold more D1.1 than normal tissue. These findings suggest that one result of the weaver mutation is prolonged expression of D1.1. We speculate that the D1.1 ganglioside might be involved in adhesive interactions that regulate the timing of granule-cell migration from the EGL. The prolonged expression of D1.1 could be responsible, in part, for the failure of granule-cell migration in the weaver cerebellum. PMID- 3049940 TI - Laboratory and epidemiologic assessment of a recent influenza B outbreak. AB - A viral surveillance system in Nashville detected an outbreak of influenza B that occurred between January and March 1986. Paired sera from 32 individuals with culture-documented influenza B illness were tested using three serologic assays. Enzyme-linked immunosorbent assay (ELISA) using purified hemagglutinin neuraminidase and plaque neutralization detected a seroresponse in 69% and 66% of these individuals, respectively. These assays were superior to hemagglutination inhibition, which detected a 41% seroresponse. ELISA was perferred because of cost and ease of performance. A group of 286 individuals, aged 1-65 years, was studied more extensively including serologic assessment before and after the influenza B outbreak. Historical information and viral throat cultures were obtained from those with influenza-like illness during the epidemic. An influenza B infection rate (seroresponse and/or positive culture) of 31% and illness rate (infection with flu-like symptoms during the epidemic period) of 13% was demonstrated using these methods. Pre-epidemic mean serum ELISA IgG titers were lower in those with, versus those without, evidence of subsequent influenza B illness (1,541 vs. 4,311, P = .0026). Children less than or equal to 15 years of age were infected more frequently than adults (44% vs. 28%, P = .04). Fever greater than or equal to 101 degrees F was reported more frequently with influenza B than non-B illness (43% vs. 18%, P = .03). These data are useful in preparing for future epidemiologic studies of influenza B and demonstrate the value of and need for standardization of ELISA as a serologic assay for influenza B. PMID- 3049943 TI - Therapeutic applications of antiidiotypic antibodies. PMID- 3049944 TI - Treatment of epidemic Kaposi's sarcoma with a combination of interferon-alpha 2b and etoposide. AB - A prospective clinical trial of concomitant interferon-alpha 2b and etoposide was conducted in 24 previously untreated patients with epidemic Kaposi's sarcoma. Eight of 21 evaluable patients (38%) achieved either a complete response (1 patient) or a partial response (7 patients). None of the responders had a prior history of opportunistic infection. Hematologic toxicity was severe, and 8 patients developed an opportunistic infection. The combination of interferon alpha 2b and etoposide has modest activity, but no additive or synergistic activity was evident in the dose and schedule utilized in this study. The exact role for interferon-alpha in epidemic Kaposi's sarcoma, both as a single agent and in combinations, remains to be determined. PMID- 3049945 TI - Immunoreactive epidermal growth factor receptors in neuritic plaques from patients with Alzheimer's disease. AB - Alzheimer's disease (AD) is characterized neuropathologically by the presence of neuritic plaques (NP) in cerebral cortex and hippocampus, as well as intraneuronal neurofibrillary tangles and granulovacuolar degeneration. The etiology of plaque formation has remained obscure, but morphologically NP are known to contain amyloid cores surrounded by astrocytes and degenerating neurons. Although growth factors are important in growth, differentiation and regrowth in response to injury, studies relating growth factors to AD have been lacking. Epidermal growth factor (EGF) plays an important role outside the central nervous system (CNS) through interaction with its specific receptor, EGF-R. Using an antibody to EGF-R (three-step immunoperoxidase staining) in conjunction with fluorescence staining, we found that the majority of NP from patients with pathologically confirmed AD as well as those few NP in the normal aging brain showed intense EGF-R immunoreactivity. Specific staining was seen at the periphery of plaques but not in the central amyloid core. Tissue sections from AD cases were also reacted with antibodies to both glial fibrillary acidic protein (GFAP) and paired helical filaments (PHF) in an attempt to identify which component of the NP was reactive for EGF-R. The antibody to PHF densely stained the periphery of NP but not the central core in a majority of NP. The antibody to GFAP stained a few reactive astrocytes that bordered plaques in only a small proportion of all plaques present. We conclude that the neuron and its processes although not exclusively may be the site of EGF-R immunoreactivity. An EGF/EGF-R system within the CNS may play an important part in scar formation in response to neuronal injury and death or it may function as a trophic factor important in axonal or dendritic sprouting. It is also possible that EGF could serve as a neurotransmitter/neuromodulator in the CNS. PMID- 3049946 TI - Immunoperoxidase labelling of rat brain sections with sera from patients with paraneoplastic cerebellar degeneration and systemic neoplasia. AB - Sera from patients with systemic cancer found by immunofluorescence staining to have antibodies to human cerebellar cell populations were reacted with vibratome sections of rat cerebellum and examined by peroxidase-antiperoxidase (PAP) methods. Seven patients with clinically or pathologically confirmed paraneoplastic cerebellar degeneration and two neurologically normal patients with high titers of anticerebellar antibodies were studied. Sera from all antibody-positive patients, but not from controls, produced intense staining of brain sections. Sera from patients with ovarian adenocarcinoma reacted predominantly with Purkinje cells and neurons within brainstem nuclei. Sera from patients with oat cell carcinoma and one patient with ductal carcinoma of the breast produced nuclear and cytoplasmic staining of neurons throughout the central nervous system. Serum from a patient with Hodgkin's disease labeled the peripheries of Purkinje cells and Golgi II cells. Serum from a patient with mixed mesodermal sarcoma of the ovary labeled Purkinje cells, basket cells, and scattered astrocytes. Staining of extraneural tissues was not observed. This study confirms the presence of antineural antibodies in patients with systemic neoplasia with and without paraneoplastic cerebellar degeneration and suggests that the antigens recognized by this antibody response may vary with the associated neoplasm. PMID- 3049947 TI - Distribution of enkephalin-immunoreactive nerve fibres and terminals in the region of the nucleus basalis magnocellularis of the rat: a light and electron microscopic study. AB - This investigation was carried out on the distribution of enkephalin-containing nerve fibres and terminals in the region of the nucleus basalis magnocellularis (NBM) of the rat. At the light microscope (LM) level, enkephalin-immunoreactive sites and endogenous choline acetyltransferase (ChAT) were demonstrated by employing the two-colour immunoperoxidase staining technique, using highly specific monoclonal antibodies against enkephalin and ChAT. A pharmacohistochemical procedure to reveal acetylcholinesterase (AChE) synthesizing neurons combined with the peroxidase-antiperoxidase (PAP) immunocytochemical technique to detect endogenous enkephalins, provided ultrastructural data on the relationships of neuronal elements containing AChE and enkephalins in the region of the NBM. At the LM level, cholinergic neurons of the NBM were surrounded by a dense network of enkephalin-immunoreactive nerve fibres. Electron microscopic (EM) observations of histochemically characterized structures, that were first identified in the LM, revealed that intensely AChE stained structures in the region of the NBM received sparse synaptic inputs from enkephalin immunoreactive terminals. Synaptic inputs of immunoreactive terminals onto intensely AChE-stained neuron cell bodies were not detected. Synaptic contacts onto proximal AChE-positive dendrites were sparse, but the density increased on more distal regions of the dendrites. All immunoreactive boutons studied established symmetrical synaptic contacts with AChE-positive post synaptic structures. The pattern of the synaptic input to these cells differs strikingly from that onto typical globus pallidus neurons. The perikarya and dendrites of the latter neurons were characteristically ensheathed in immunoreactive synaptic boutons. Results are consistent with the view that enkephalin-like substances in the rat might be synaptic transmitters or neuromodulators in the area of the NBM and that cholinergic neurons of the NBM (Ch4) are integrated into the circuitry of the basal ganglia. Enkephalins may play an important role regulating the extrinsic cholinergic innervation of the neocortex. PMID- 3049948 TI - Allogeneic transplantation versus intensive chemotherapy in first-remission acute leukemia: is there a "best choice"? PMID- 3049949 TI - A comparison of marrow transplantation with chemotherapy for adults with acute leukemia of poor prognosis in first complete remission. AB - From July 1980 to November 1985, 109 patients with acute myelogenous and lymphoblastic leukemia who had reached a complete remission (CR) following induction treatment were assigned to a study comparing marrow transplantation with chemotherapy as a postremission treatment. Sixty-nine patients did not have a human leukocyte antigen (HLA)-identical donor, and therefore served as chemotherapy controls; 40 patients had HLA-identical donors, and therefore were assigned to the transplant arm. Of these, 23 were transplanted in first remission and 17 were not. Ten of these 17 were subsequently transplanted in relapse. Initially, only patients with poor prognosis determined by a predictive model were entered into the study. Subsequently, patients with moderately poor prognosis were admitted. Comparing the chemotherapy group with the patients transplanted in first CR, significant control of leukemia relapse in transplanted patients was seen in the subgroup with acute myelogenous leukemia (AML) (P less than .1), and acute lymphoblastic leukemia (ALL) (P less than .01), in the poor (P = .01) and intermediate subgroup (P = .01), and in the good-prognostic groups (P = .05). The survival was affected significantly in only the poor and intermediate subgroups. The use of predictive models might help to select patients for whom bone marrow transplantation is appropriate in first remission and those for whom bone marrow transplantation can be left as the initial treatment of relapse. Predictive models could further be helpful in comparing studies performed at different transplant centers. PMID- 3049950 TI - Successful marrow transplantation for acute myelocytic leukemia following therapy for Hodgkin's disease. AB - Five patients with acute myelocytic leukemia (AML) after combined modality therapy for Hodgkin's disease (HD) were treated with cyclophosphamide and busulfan followed by bone marrow transplantation (BMT). Four patients received allogeneic transplants from histocompatibility locus antigen (HLA)-compatible siblings and the fifth patient received an autologous marrow treated with 4 hydroperoxycyclophosphamide. Two patients died of complications of acute graft-v host disease (GVHD) despite prophylaxis with either low-dose cyclophosphamide or cyclosporine. The remaining three patients were alive and disease-free 382, 617, and 620 days after transplant. These initial results are encouraging and more patients with treatment-related AML need to be evaluated with both allogeneic and autologous BMT to fully elucidate the potentially curative role of this intensive therapy in an otherwise fatal hematologic malignancy. PMID- 3049951 TI - Regimen-related toxicity in patients undergoing bone marrow transplantation. AB - Bone marrow transplantation is associated with significant morbidity and mortality, some of which is due to high-dose chemoradiotherapy. In order to quantitate toxicity that was felt to be due to the preparative regimen (termed regimen-related toxicity [RRT]), a system was developed in which toxicities were graded from 0 (none) to 4 (fatal). One hundred ninety-five patients who underwent marrow transplantation for leukemia were studied retrospectively to determine whether toxicities that were clinically felt to be due to the preparative regimen were influenced by other factors such as disease status, graft-versus-host disease (GVHD) prophylaxis, and allogenicity. All patients developed grade I toxicity in at least one organ, and 30 developed grades III-IV (life-threatening or fatal) RRT. RRT was more common in relapsed patients v remission patients (P = .04), in those receiving 15.75 Gy total body irradiation (TBI) v 12.0 Gy TBI (P = .028), and in those receiving allogeneic marrow v autologous marrow (P = .0029). Autologous marrow recipients did not develop grades III-IV toxicity in this study. A multivariate analysis controlling for autologous marrow grafting showed that the dose of TBI was the only statistically significant predictor of grades III-IV RRT. Those patients who developed grade III RRT were unlikely to survive 100 days from transplant, though not all deaths could be attributed to RRT. Patients who developed grade II toxicity in three or more organs were more likely to die within 100 days than those developing grade II toxicity in two or less organs (P = .0027). This system was generally able to distinguish RRT from other toxicities observed in marrow recipients. PMID- 3049952 TI - Recombinant interferon alfa-2a in metastatic renal cell carcinoma: assessment of antitumor activity and anti-interferon antibody formation. AB - Twenty-one patients with advanced, measurable, renal cell carcinoma (RCC) were administered recombinant interferon alfa-2a (rIFN-alpha 2a) (Roferon-A; Roche Laboratories, Nutley, NJ) intramuscularly beginning at 3 x 10(6) units and escalating to 36 x 10(6) units, 5 d/wk for a total induction period of 14 weeks. rIFN-alpha 2a antibody production was measured using an enzyme immunoassay (EIA). Those sera found to be positive for presence of antibody by the EIA were tested for the presence of neutralizing antibodies (NA) by an antiviral neutralization bioassay (ANB). All patients were evaluable for toxicity, and 19 were evaluable for response and for incidence of antibody formation. Five patients (26%; 95% confidence interval, 6% to 46%) had complete responses (CR) or partial responses (PR) with a median duration of 283 days. An additional ten patients (53%) had minor tumor regressions with a median duration of 86 days. Fifty-one percent of evaluable patients are alive at 18.6 months. Antibodies to rIFN-alpha 2a as measured by the EIA, were detected in 12 (63%) patients. NA were measured in the serum of six (50%) of those EIA-positive patients. Overall, six of 19 patients (32%) developed NA. Median time to the development of antibody as measured by EIA or NA was 8 and 14 weeks, respectively. Median NA titer was 1,200 IFN neutralizing U/mL. NA-positive and -negative patients had a median duration of response of 13.7 v 9.9 months, and survival of greater than 21.3 v 18.3 months, respectively. Clinical toxicity was mild and not therapeutically limiting. Autoantibody production (ANA, rheumatoid factor [RF], Coombs' direct/indirect) occurred in both NA-positive and -negative patients. The clinical significance of the antibodies to rIFN-alpha 2a and the associated autoantibody formation remain unclear; however, presence of antibody was not associated with adverse clinical sequelae. PMID- 3049953 TI - A randomized multicenter trial comparing mitoxantrone, cyclophosphamide, and fluorouracil with doxorubicin, cyclophosphamide, and fluorouracil in the therapy of metastatic breast carcinoma. AB - Three hundred thirty-one women with metastatic breast cancer were randomized to receive combination chemotherapy with either cyclophosphamide, Novantrone (mitoxantrone; Lederle Laboratories, Wayne, NJ), and fluorouracil (CNF) or cyclophosphamide, Adriamycin (doxorubicin; Adria Laboratories, Columbus, OH), and fluorouracil (CAF). Patients could not have had prior chemotherapy, although adjuvant chemotherapy was acceptable. Initial doses were 500 mg/m2 of cyclophosphamide and 500 mg/m2 of fluorouracil with either 10 mg/m2 of mitoxantrone or 50 mg/m2 of doxorubicin, administered intravenously (IV) on day 1 and repeated every 3 weeks. There were no statistically significant differences in pretreatment or prior therapy characteristics between the groups. For patients assigned to the CNF and CAF groups, respectively, 25 (18%) were premenopausal, 39 (40%) were estrogen receptor (ER) negative, 39 (38%) had a disease-free interval less than 1 year, and 24 (26%) had received prior adjuvant chemotherapy. All patients were compared for response rate, duration of response, time to progression or death, time to treatment failure (TTF), and survival. None of these parameters were statistically significant favoring one regimen over the other. The response rate (complete [CR] and partial response [PR]) was 29% for the CNF group (95% confidence interval of 22% to 37%) and 37% for the CAF group (95% confidence interval of 29% to 45%). The median response duration and TTF were 171 days and 125 days for the CNF group and 254 days and 147 days for the CAF group, respectively. The median survival times for the CNF group and the CAF group were 377 and 385 days, respectively. The major dose-limiting toxicity for both regimens was leukopenia, manifested as granulocytopenia. The incidence of stomatitis/mucositis was 10% in the CNF group and 19% in the CAF group. Alopecia occurred in 49% of CNF patients (severely for 4%) and in 86% of CAF patients (severely for 39%). Nausea/vomiting occurred in 80% of CNF patients and in 81% of CAF patients; the degree of severity was also comparable. There was significantly less cardiotoxicity observed in the CNF group compared with the CAF group. Although CNF is somewhat less effective in overall response rate, survival curves are identical. CNF can be offered to patients who reject anthracycline-containing regimens because of fear of alopecia. PMID- 3049954 TI - Fluorouracil: biochemistry and pharmacology. AB - Fluorouracil (5FU) is still considered the most active antineoplastic agent in the treatment of advanced colorectal cancer. The drug needs to be converted to the nucleotide level in order to exert its effect. It can be incorporated into RNA leading to interference with the maturation of nuclear RNA. However, its conversion to 5-fluoro-2'deoxy-5' monophosphate (FdUMP) leading to inhibition of thymidylate synthase (TS) and subsequently of DNA synthesis, is considered to be its main mechanism of action. In the presence of a folate cofactor a covalent ternary complex is formed, the stability of which is the main determinant of the action of 5FU. Resistance against 5FU can be mainly attributed to aberrations in its metabolism or to alterations of TS, eg, gene amplification, altered kinetics in respect to nucleotides or folates. Biochemical modulation of 5FU metabolism can be applied to overcome resistance against 5FU. A variety of normal purines, pyrimidines, and other antimetabolites have been studied in this respect, but only some of them have been clinically successful. Delayed administration of uridine has recently been shown to "rescue" mice and patients from toxicity, while pretreatment with leucovorin is the most promising combination to enhance the therapeutic efficacy. 5FU is frequently administered in an intravenous (IV) injection, and shows a rapid distribution and a triphasic elimination. The nonlinearity of 5FU pharmacokinetics is related to saturation of its degradation. Continuous infusion of 5FU led to different kinetics. Regional administration, such as hepatic artery infusion, offers a way to achieve higher drug concentrations in liver metastases and is accompanied by lower systemic concentration. The current status of the biochemical and pharmacokinetic data is reviewed. PMID- 3049955 TI - Rat astroglial somatomedin/insulin-like growth factor binding proteins: characterization and evidence of biologic function. AB - Specific binding proteins (BPs) to somatomedin/insulin-like growth factors (Sm/IGFs) have been identified in conditioned media from a variety of cells in culture. By affinity cross-linking using disuccinimidyl suberate, we have covalently cross-linked radiolabeled somatomedin-C/insulin-like growth factor I (Sm-C/IGF I), insulin-like growth factor II (IGF II) and insulin to BPs in conditioned medium (CM) from cultured astroglial cells derived from cerebral cortices of neonatal rats. Two species of radiolabeled Sm/IGF BP complexes of 40,000 Da (40K) and 45K were identified. Competition with unlabeled Sm-C/IGF I and IGF II demonstrated that the BPs in each complex have similar affinities for Sm-C/IGF I and IGF II. The BP in the 45K complex was about 5-fold more sensitive to competition with unlabeled Sm/IGFs than the BP in the 40K complex, suggesting that it either has a higher affinity for Sm/IGFs or is less abundant. Evidence that the BPs in each complex are distinct includes the following findings: (1) insulin competed with Sm/IGF for binding to the 45K complex, but not the 40K complex, and (2) the BP in the 40K complex, but not the 45K complex, was recognized by antibodies raised against a BP purified from CM of buffalo rat liver (BRL) 3A cells. Growth hormone did not affect the apparent secretion of either BP. The binding activity of both BPs was retained after mild heat treatment, changes to extremes of pH (2-10), and prolonged storage at -20 degrees C, but was destroyed after heating to higher temperatures (80 degrees C and greater), reduction, and proteolytic treatment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3049956 TI - A monoclonal antibody to the insect prothoracicotropic hormone. AB - The prothoracicotropic hormone (PTTH) is an insect cerebral peptide that stimulates the prothoracic glands to produce the steroid hormone ecdysone thus initiating molting and metamorphosis. "Big" PTTH, one of several molecular forms of the neurohormone, was isolated from brains of the tobacco hornworm Manduca sexta, and fractionated by high-pressure liquid chromatography (HPLC) for use in antibody production. A murine polyclonal antiserum and a monoclonal antibody (MAb) have been generated using this highly purified preparation of big PTTH. Antisera and hybridoma supernatants were screened with an indirect, brain whole mount immunocytological assay, and antibody specificity was confirmed by immunocytological, ELISA, and functional criteria. In brain whole-mount preparations, the MAb (A2H5) and antiserum specifically immunostained the lateral protocerebral neurosecretory cells (L-NSC III), the prothoracicotropes, which produce PTTH. This immunostaining was blocked by preadsorbing the antibodies with big PTTH. Analysis of the elution of HPLC-fractionated big PTTH with an in vitro bioassay for the neurohormone and an ELISA employing the A2H5 MAb resulted in peaks of activity that were superimposable. Finally, the antiserum and A2H5 MAb inhibited big PTTH activation of the prothoracic glands to synthesize ecdysone in the in vitro bioassay for the neurohormone. With these specific antibodies, the organization of the PTTH neuroendocrine axis has been defined. It is now evident that both of the peptidergic neurons that comprise the L-NSC III are prothoracicotropes, and that the corpora allata are the neurohemal organs for the release of big PTTH into the hemolymph. This study indicates that these specific antibodies will be useful in investigations of numerous aspects of the biology of this cerebral neuroendocrine axis. PMID- 3049957 TI - Histological observations on the healing of a periodontal defect five years after an autogenous bone graft. PMID- 3049958 TI - Quantitative measurement of renal perfusion following transplant surgery. AB - We developed an easily implemented clinical procedure for quantitative perfusion measurements in transplanted kidneys using intravenously administered [99mTc]DTPA and the tracer fractionation technique. F = Ak(T)/0 integral of T [Aa(t)/Va] dt, where F = renal blood flow, Ak(T) = DTPA activity in kidney at time = T, Va = ultrasonographically measured femoral artery segment volume, T = time postinjection of F determination, and Aa(t) = time course of DTPA activity in femoral artery segment. The technique was applied to a group of 80 studies in 35 patients in whom an independent clinical determination of transplant function was available. Blood flow (units of ml/min) measured 439 +/- 83 in normally functioning transplants, 248 +/- 63 in transplants with acute tubular necrosis, 128 +/- 62 in transplants with rejection, and 284 +/- 97 in transplants with cyclosporine toxicity. These preliminary results indicate potential usefulness of this method in the evaluation of renal function following transplant surgery. PMID- 3049959 TI - Comparison of the biodistribution of gadolinium-153 DTPA and technetium-99m DTPA in rats. AB - Twenty-three mature Sprague-Dawley male and female rats were simultaneously injected with trace quantities of [153Gd]DTPA and [99mTc]DTPA and 0.5 mmol/kg of nonradioactive gadolinium DTPA. Rats were killed at 1 min, 5 min, 10 min, 15 min, and 30 min after the intracardiac bolus injection. The heart, lungs, liver, brain, kidney, and blood were excised and counted in a well-counter to determine the amount of the injected material in each organ and blood. In order for the percent of total injected activity to be determined, a technique was developed which allowed discrimination of the 140 keV gamma-ray of 99mTc from sum peaks of 153Gd when the latter is counted in a well-counter with 4 pi geometry. Although the distribution of the two DTPA compounds was qualitatively similar, statistical analysis indicated that the amount of 99mTc deposited in the lungs was higher than 153Gd (p = 0.03), the amount of 99mTc deposited in the kidneys was lower than 153Gd (p = 0.0004) and the amount of 99mTc in the blood was higher than 153Gd (p = 0.0022). This may be due to the greater binding of [99mTc]DTPA or its minor impurities to plasma proteins. PMID- 3049960 TI - Ultrasonic guidance for internal mammary lymphoscintigraphy. PMID- 3049961 TI - A radiopharmaceutical for the study of the liver: 99mTc-DTPA-asialo-orosomucoid. I: Radiochemical and animal distribution studies. PMID- 3049962 TI - A radiopharmaceutical for the study of the liver: 99mTc-DTPA-asialo-orosomucoid. II: Human dynamic and imaging studies. PMID- 3049963 TI - Does seasonal employment in grain elevators increase nonspecific airways responsiveness? AB - Grain handling and increased airways responsiveness (AR) have been independently associated with an accelerated decline in forced expiratory volume in 1 sec. We performed methacholine inhalation bronchial challenge tests in 45 river port grain handlers during layoff and again during employment to determine whether short-term exposure increased AR. To assess bias, due to seasonal/temporal influences, AR was also measured in 56 nonexposed men of similar age and socioeconomic status. AR, slightly higher among laid-off grain handlers than the comparison group, fell more among grain handlers during employment than among the comparison group during the same time period. Our results do not support the hypothesis that seasonal exposure to grain dust increases AR. PMID- 3049965 TI - Have we overlooked important cohorts for follow-up studies? Report of the Chemical Industry Institute of Toxicology Conference of World War II-era Industrial Health Specialists. AB - During the World War II era large segments of the industrial work force suffered heavy exposure to toxic materials. Often there followed episodes of acute toxic illness, which may have been precursors of other effects, as yet unidentified. Yet adequate follow-up studies to document the subsequent health history of these populations have not been undertaken. Such studies should be initiated while knowledgeable persons and necessary records may still be found. Existing epidemiologic data bases, constructed after considerable effort, must be preserved in an archive for the benefit of future researchers. It is essential to identify and characterize important present-day cohorts now, although their experiences of interest to scientists may not occur for many years. PMID- 3049964 TI - Allergic sensitization to a non-bisphenol A epoxy of the cycloaliphatic class. AB - This is the first report of sensitization to a cycloaliphatic epoxy. Allergic contact dermatitis occurred in an electron microscopist after exposure to the cycloaliphatic epoxy, vinyl cyclohexene diepoxide. Cycloaliphatic epoxies are not based on the diglycidyl ether of bisphenol A (DGEBA), presently the epoxy of greatest commercial usage. However, non-DGEBA epoxies are finding new applications in the semiconductor and aerospace industries and will likely gain in importance as a cause of occupational allergic dermatitis. Latex or polyvinyl chloride gloves did not protect the reported patient from percutaneous absorption and elicitation of allergic dermatitis. PMID- 3049966 TI - Quantitating bedside diagnosis: clinical evaluation of ascites. AB - The authors prospectively evaluated the operating characteristics of the history and physical examination for ascites in a broad spectrum of hospitalized patients. The overall clinical evaluation produced a positive likelihood ratio = 37.7-83.3 when suggestive of ascites, a likelihood ratio = 2.23-3.42 when intermediate, and a negative likelihood ratio = 0.77-0.90 when not suggestive of ascites. Patients' perceptions of increased abdominal girth (positive likelihood ratio = 4.16) or recent weight gain (positive likelihood ratio = 3.20) increased the likelihood of ascites. The absence of subjective ankle swelling (negative likelihood ratio = 0.10) or increased abdominal girth (negative likelihood ratio = 0.17) decreased the likelihood of ascites. The positive likelihood ratios for a fluid wave = 9.6 and shifting dullness = 5.76 favored ascites, while the absence of bulging flanks (negative likelihood ratio = 0.12) or peripheral edema (negative likelihood ratio = 0.17) favored ascites the least. Thus, a routine history and physical examination are quantitatively useful in the clinical evaluation of ascites. PMID- 3049967 TI - The assessment of diagnostic tests: a comparison of medical literature in 1982 and 1985. AB - To determine whether improvements have occurred since a survey of the 1982 literature assessing diagnostic tests, the authors evaluated all English-language articles that assessed clinical diagnostic tests in abridged Index Medicus journals in 1985, and that had the terms sensitivity and specificity in the title, abstract, or key words. The 89 articles were assessed against seven methodologic criteria, including use of a well-defined "gold standard," clearly defined test interpretation, blinding, clear data presentation, correct use of sensitivity and specificity, calculation of predictive values, and consideration of prevalence. In comparisons of 1985 vs. 1982 articles, there were significant improvements in five of the seven criteria. For example, the proportion of articles using a well-defined "gold standard" rose from 68% to 88%. Overall, the frequency of papers demonstrating five or more of the seven criteria increased from 26% to 47%. However, predictive values were discussed in only 54% of the articles without, necessarily, consideration of the influence of prevalence as well. This study raises the concern that while the concepts of sensitivity and specificity are now accepted, predictive values remain less well understood. Although there has been an improvement in the assessment of diagnostic tests in published research, attention to accepted methodologic standards is still needed on the part of researchers, reviewers, and editors. PMID- 3049969 TI - Reference test errors bias the evaluation of diagnostic tests for ischemic heart disease. PMID- 3049968 TI - Patients' participation in medical care: effects on blood sugar control and quality of life in diabetes. AB - To maximize disease control, patients must participate effectively in their medical care. The authors developed an intervention designed to increase the involvement of patients in medical decision making. In a 20-minute session just before the regular visit to a physician, a clinic assistant reviewed the medical record of each experimental patient with him/her, guided by a diabetes algorithm. Using systematic prompts, the assistant encouraged patients to use the information gained to negotiate medical decisions with the doctor. A randomized trial was conducted in two university hospital clinics to compare this intervention with standard educational materials in sessions of equal length. The mean pre-intervention glycosylated hemoglobin (HbA1) values were 10.6 +/- 2.1% for 33 experimental patients and 10.3 +/- 2.0% for 26 controls. After the intervention the mean levels were 9.1 +/- 1.9% in the experimental group (p less than 0.01) and 10.6 +/- 2.22% for controls. Analysis of audiotapes of the visits to the physician showed the experimental patients were twice as effective as controls in eliciting information from the physician. Experimental patients reported significantly fewer function limitations. The authors conclude that the intervention is feasible and that it changes patient behavior, improves blood sugar control, and decreases functional limitations. PMID- 3049970 TI - Measurement of severity of illness and the Medicare prospective payment system: state of the art and future directions. PMID- 3049972 TI - Depression in the elderly: the role of the primary care physician in management. PMID- 3049974 TI - The Registry of the International Society for Heart Transplantation: fifth official report--1988. PMID- 3049971 TI - Depression in primary care: DSM-III diagnoses and other depressive syndromes. PMID- 3049975 TI - Heart transplantation in patients over age fifty-five years. AB - The age limit for heart transplantation remains undefined. The shortage of available donors coupled with fears of increased morbidity and mortality in older patients has until recently resulted in a limited application of heart transplantation in patients over age 50 years. In 1986, however, data from the International Heart Transplant Registry demonstrated that 25% of all patients undergoing this procedure were over age 55 years. This study reviews our experience with 30 consecutive patients who underwent heart transplantation over a 15-month period. There were eight patients over age 55 years (group 1) and 22 patients under age 55 years (group 2). We compared the hospital course and incidence of infection and rejection and other complications after heart transplantation between the two groups. Carefully selected patients between ages 55 and 60 years can undergo transplantation with similar expectations to younger patients for survival, complications, and rehabilitation, including employability. Caution is warranted in extrapolating these optimistic data to patients older than age 50 years. PMID- 3049973 TI - Post-stroke depression in the elderly. PMID- 3049976 TI - Heart transplantation in elderly patients. AB - During 1985 to 1986, 57 orthotopic heart transplantations have been performed at the University of Minnesota, Minneapolis. All patients received triple-drug immunosuppressive therapy of cyclosporine, azathioprine, and prednisone. Twenty three patients were aged 55 years or older (mean age 58.0 +/- 2.6 years); 34 patients were under the age of 55 years (mean age 39.1 +/- 13.2 years). The initial in-hospital stay averaged 13 +/- 4 days in the older group and 16 +/- 13 days in the younger group. Perioperative mortality was similar in the two groups. The incidence of cerebrovascular accident was similarly low in the two groups, whereas incidence of steroid-induced diabetes (17% versus 9%) and significant osteoporosis (13% versus 3%) was significantly higher in older patients. The probability of survival and the actuarial freedom from rejection were identical in the two groups with 1-year survival of 96% and 94% of patients free of rejection at 12 months. Although the incidence of infection was not significantly higher in older patients (0.82 episode per patient) than in younger patients (0.78 episode per patient), life-threatening infections (Cryptococcus meningitidis, disseminated herpes simplex) were observed only in older patients. These data suggest that (1) heart transplantation is a valid therapeutic option even in elderly patients with end-stage heart failure, (2) it can be performed in selected patients at no increased operative risk with excellent long-term survival, (3) however, older patients are at higher risk for serious infections and for developing steroid-related complications. PMID- 3049977 TI - Simultaneous retrieval of the heart and liver from a single donor: an evaluation through preservation and transplantation. AB - The simple, safe, and feasible procurement technique for the heart and liver with no warm ischemic time is reported. Fifteen mongrel dogs were used to form two recipients and one donor combination in each experiment. A midline incision is extended from the suprasternal notch to the pubis, and a catheter is advanced into the aortic root by means of the brachiocephalic artery for monitoring systemic arterial pressure and later for coronary vascular washout with a cold cardioplegic solution. Liver mobilization is carried out first when the core temperature of the liver reaches 27 degrees C, obtained with ice slush in the abdominal cavity. As the core temperature of the liver reaches 20 degrees to 22 degrees C, the aorta and inferior vena cava are clamped just above the diaphragm. After excision of the liver a second team harvests the heart while it continues to beat. The heart and liver were transplanted orthotopically after simple preservation into a cold solution for 12 and 6 hours, respectively. The maximum survival time was 7 hours in heart transplantation and 16 days in liver transplantation. Our method is a simple, safe, feasible technique for acquiring the heart and liver or other visceral organs for transplantation and may have broad clinical application. PMID- 3049979 TI - Systolic anterior motion of the mitral valve as a manifestation of heart transplant rejection. AB - A 65-year-old white man with acute cardiac allograft rejection had a diagnosis made on the basis of clinical presentation and endomyocardial biopsy. The echocardiogram showed systolic anterior motion of the mitral valve during the episode of rejection. There was no systolic anterior motion on the echocardiogram that was done either before or after the episode of rejection. PMID- 3049978 TI - Use of power spectral analysis of respiratory sinus arrhythmia to detect graft rejection. AB - Some evidence has suggested that graft rejection in heart transplant recipients is accompanied by sinus node dysfunction. To test this hypothesis the electrocardiograms of 21 orthotopic heart transplant recipients receiving cyclosporine were recorded at regularly scheduled biopsies. This resulted in 105 good quality recordings of at least 10 minutes each. All exhibited normal sinus rhythm. These recordings were then processed through patented digital routines that calculated the power spectrum of the RR intervals. All recordings exhibited Mayer wave and respiratory sinus arrhythmia (RSA) periodicities. Peak spectral power of RSA (PSP-RSA) demonstrated good repeatability at 2 weeks (r = 0.81). A paired t test comparing the baseline PSP-RSA in eight subjects who experienced rejection episodes with their individual average PSP-RSA during periods of rejection revealed a significant decrease (p = 0.039). As a global measure, PSP RSA greater than 1 (natural logarithmic scales) demonstrated a sensitivity of 1.0, specificity of 0.42, positive predictive value of 0.22, and negative predictive value of 1.0. These tentative results demonstrate that PSP-RSA may be a sensitive, noninvasive marker for graft rejection of heart transplant patients receiving cyclosporine for immunosuppression. A larger, more extensive study needs to be conducted to confirm these results. PMID- 3049980 TI - Chronic rejection in human heart transplantation. AB - The more prevalent complication in patients with a long survival rate after heart transplantation is chronic rejection, which was studied in a series of 80 necropsies and five cardiac grafts surgically removed for retransplantation after chronic rejection. In the material obtained at necropsy, 11 of 14 patients with a survival rate of more than 6 months died from chronic rejection. Clinically, the usual manifestation was heart failure. Anatomic angiograms were performed in several cases. They demonstrated narrowing and nonopacification of small coronary arteries, often accompanied by thrombosis and ischemic complications. The histologic study detected three types of rejection. (1) The more typical rejection is observed after 6 months. It is characterized by a stenosing fibrous endarteritis. (2) Another type of rejection occurs earlier and is associated with acute rejection; its anatomic substratum is an inflammatory panarteritis. (3) This type of rejection is accompanied by large atheromatous deposits. The significance and pathogenesis of these lesions are discussed in correlation with their clinical context and with the electron microscopic observations. PMID- 3049982 TI - Myocardial calcification after orthotopic heart transplantation. AB - Two cases with a remarkable similarity in their clinical and histopathologic findings are reported. Both cases involved myocardial calcification found in biopsy material obtained after orthotopic heart transplantation. Myocardial calcification after heart transplantation so far has been described in only one case in the medical literature and thus appears to be a rare entity. Its occurrence is associated with a certain constellation of clinical situations, which include repeated episodes of acute rejection, temporary uremia, periods of septicemia, alcoholism, and cyclosporine and/or steroid therapy. PMID- 3049981 TI - Technique of clinical double-lung transplantation. AB - Clinical double-lung transplantation has now been successfully performed at the University of Minnesota, Minneapolis. Donor care and operation are similar to heart-lung donor management. The most important parts of the recipient operation are to remove the lungs without injury to the phrenic, vagal, or recurrent nerves and to ensure hemostasis. In addition, the integrity of the tracheal anastomosis should be ensured by an omental wrap. The operative technique, developed and modified from our laboratory experience, is described. PMID- 3049984 TI - Handling bioptomes. PMID- 3049983 TI - Orthotopic heart transplantation eleven years after left pneumonectomy. AB - Orthotopic heart transplantation was performed in a 21-year-old medical student 11 years after left pneumonectomy for a rhabdomyosarcoma. The cardiomyopathy was the result of the administration of doxorubicin (Adriamycin). The surgical procedure was largely facilitated as a result of an in-hospital donor and the absence of major adhesions. The early postoperative course was mainly uneventful. The patient is doing well 9 months after operation, without any episode of rejection or infection. PMID- 3049985 TI - Cardiopulmonary preservation. PMID- 3049986 TI - Cardiopulmonary preservation. PMID- 3049987 TI - Quality of life after transplantation. PMID- 3049988 TI - Indications for heterotopic heart transplantation and report on two patients. PMID- 3049989 TI - Laboratory diagnosis of rabies. PMID- 3049990 TI - Destruction or reconstruction? PMID- 3049992 TI - Recurrent osteoblastoma of the mandible: report of a case. PMID- 3049991 TI - Short-term storage of freshly harvested bone. AB - An in vitro study was designed to test effects of various graft storage media on glucose metabolism and collagen synthesis of embryonic chick tibiae, using these variables as indices of bone cell viability. Normal saline solution, distilled water, and 5% dextrose in lactated Ringer's solution were evaluated after a 5 hour incubation and again after a 3-day recovery period in a complete culture medium. The study suggests that bone grafts may be stored in normal saline solution or 5% dextrose in lactated Ringer's solution for up to 5 hours. Normal saline solution is recommended because of fewer deleterious effects. Distilled water should not be used as a storage medium for bone grafts. PMID- 3049993 TI - Mesenchymal chondrosarcoma of the maxilla: report of a case. AB - A case of mesenchymal chondrosarcoma of the maxilla in a 68-year-old patient is presented. The gross and histologic appearance of the tumor is described and the literature is reviewed. PMID- 3049994 TI - The use of tissue expanders in the correction of avulsive injuries of the mandible: report of two cases. AB - Two cases illustrating successful use of tissue expanders to correct both soft and bony defects of the facial region are presented. The tissue expanders allow the development of a flap that has the same color and texture as the lost tissue. It also precludes the need of a distantly developed flap with its associated morbidity. The highly vascular fibrous capsule developed around the expander provides the advantage of initially greater blood supply and increased bulk for coverage of a bone graft. Although this is temporary in nature, it provides a desirable receptor bed during the critical period of bony union. The ready availability of adjacent tissue, the esthetic considerations, and the comparative reduction in morbidity when compared with use of distant flaps makes the use of tissue expanders an attractive technique in maxillofacial reconstruction. PMID- 3049995 TI - Embryonal rhabdomyosarcoma arising in the masseter muscle as a second malignant neoplasm. AB - A case is reported about a patient who was originally treated for bilateral retinoblastoma and subsequently developed an embryonal rhabdomyosarcoma in the masseter. Such patients have a genetic predisposition to a second malignancy that statistically far exceeds the rate for the general population. In addition, current treatment methods also increase the patient's susceptibility to another malignancy. This case emphasizes the necessity of maintaining a high degree of clinical suspicion in the evaluation of any lesion that may appear subsequent to the treatment of cancer in children, particularly bilateral retinoblastoma. PMID- 3049996 TI - The effect of seating pressure and powder/liquid ratio of zinc phosphate cement on the retention of crowns. AB - The effect of three different powder/liquid ratios of one zinc phosphate cement on the early retention of a standard crown on metal and tooth dies was investigated. Two different seating pressures were evaluated. The results indicate that the effect of variation in the powder/liquid was not significant. The part played by the seating force was significant, the higher seating force resulting in a greater force being required for displacement after 1 h. Greater forces of removal were required when tooth dies were used. PMID- 3049997 TI - The value of the Gothic arch tracing in the positioning of denture teeth. AB - Twenty-five subjects of three nationalities carried out Gothic arch tracings. Measurements between the side arms were compared with the upper intercuspid distances measured in the same subjects. A relationship was found which may be of value in the setting up of anterior maxillary denture teeth. PMID- 3049998 TI - A criterion for the selection of artificial posterior teeth. AB - Mandibular movement and masticatory performance were evaluated in the edentulous patients with an experimental denture using posterior artificial teeth of different occlusal schemes in order to determine the criterion for the selection of posterior artificial teeth. Lower masticatory performance was observed with non-anatomic posterior teeth than with semi-anatomic or anatomic ones regardless of the condition of residual ridge and inclination of the horizontal condylar guidance. The value of the lateral component of the masticatory cycle was closely related to the inclination of the cusp and the value of masticatory performance. This suggested strongly that the component could be a criteria for the selection of the type of the artificial posterior teeth. PMID- 3049999 TI - The effect of variation of residual ridge angle on partial denture abutment tooth movement. AB - Using a laboratory model simulating a free-end saddle partial denture situation, the effect of variation of the angulation of the crest of the residual alveolar ridge to the horizontal on movement of the abutment tooth and saddle was observed. It was found that the inclination of the residual ridge affected the direction and magnitude of abutment tooth movement. The direction of abutment tooth movement was not related to the position of the occlusal rest. However, the magnitude of saddle and abutment tooth movement is affected by the design of the clasp used. PMID- 3050000 TI - Determination of vertical dimension by hydraulic intraoral jack. AB - This paper describes the application of a hydraulic intraoral jig to the establishment of the vertical dimension of occlusion for full denture construction. The device enables the patient to establish an occlusal height which is most comfortable without the intervention or guidance of the dentist. Six patients for whom new dentures were to be constructed in the University of Otago School of Dentistry were invited to participate in the study. A conventional technique of denture construction was employed, but an additional stage was introduced after the jaw records had been taken and the casts mounted. Each patient was instructed in the use of the hydraulic jig and was asked to find a comfortable bite height in his own time. The results were compared with occlusal heights determined by conventional subjective methods. The former results proved to be more repeatable than those utilizing rest position and an average free-way space. The results are discussed and the implications outlined. Suggestions for further research using the hydraulic jig are made and the authors conclude that the hydraulic jig may be used to determine vertical dimension for full denture construction. PMID- 3050002 TI - An in vitro comparison of eight rapid streptococcal antigen detection tests. PMID- 3050003 TI - Marrow transplantation in chronic granulomatous disease: an update, with 6-year follow-up. PMID- 3050001 TI - Polysaccharide-protein conjugate vaccines for the prevention of Haemophilus influenzae type b disease. PMID- 3050004 TI - Oligohydramnios, renal insufficiency, and ileal perforation in preterm infants after intrauterine exposure to indomethacin. AB - Three preterm infants exposed antenatally to indomethacin developed a characteristic syndrome consisting of edema and hydrops with a bleeding disorder at birth, oliguric renal failure during the first 3 postnatal days, and acute pneumoperitoneum resulting from localized ileal perforation(s) at the end of the first week of life. Despite the value of indomethacin for arresting preterm labor, the physician must take into account the potential hazards of drug toxicity. PMID- 3050005 TI - Efficacy of oral sucralfate suspension in prevention and treatment of chemotherapy-induced mucositis. AB - The efficacy of orally administered sucralfate suspension in preventing and treating chemotherapy-induced mucositis was evaluated in a double-blind trial. Forty-eight children and adolescents with newly diagnosed acute nonlymphocytic leukemia were randomized to receive suspensions of either sucralfate or placebo orally every 6 hours during the first 10 weeks of intensive remission-induction chemotherapy. Patients given sucralfate suspension were less likely than subjects receiving placebo to acquire colonization with potentially pathogenic microorganisms: 14 (58%) of 24 versus 22 (92%) of 24, respectively (p = 0.008). However, no effect on preexisting colonization was noted. Subjective reporting of discomfort, objective scoring of the severity of mucositis, and the maximal percent of body weight lost during therapy were similar; 58% of patients receiving sucralfate reported no oral pain compared with 25% receiving placebo (p = 0.06). Ten episodes of gastrointestinal bleeding, 25 documented infections, and 886 days with fever were also equally distributed between sucralfate and placebo groups. We conclude that sucralfate suspension is of limited, if any efficacy, in the prevention and treatment of chemotherapy-induced mucositis. Sucralfate administration can, however, reduce acquisition of alimentary colonization with potential pathogens, perhaps by interfering with adherence to mucosal membranes. PMID- 3050007 TI - Purple versus yellow: preventing neonatal jaundice with tin-porphyrins. PMID- 3050006 TI - A randomized, placebo-controlled trial of effects of dexamethasone on hypothalamic-pituitary-adrenal axis in preterm infants. AB - As part of a blinded, randomized, placebo-controlled study of dexamethasone therapy in 27 preterm infants with bronchopulmonary dysplasia, we investigated the effect of 7 days of high-dose glucocorticoid therapy on the hypothalamic pituitary-adrenal axis. Before therapy the median basal cortisol concentration in all infants was 8.2 micrograms/dl (226 nmol/L). After stimulation with 1-24 ACTH, the serum cortisol concentration rose in all infants to a median concentration of 23.5 micrograms/dl (649 nmol/L), resulting in a median rise of 13.4 micrograms/dl (37 nmol/L). Immediately after 7 days of glucocorticoid therapy basal and peak cortisol concentrations were significantly decreased in the dexamethasone group. The rise in serum cortisol following 1-24 ACTH, however, remained equivalent in both groups. Ten days after the end of therapy basal and peak cortisol concentrations in the dexamethasone group had returned to levels equivalent to those seen in the placebo group. Weight gain was markedly diminished while the infants were receiving dexamethasone. Weight gains were, however, equivalent 10 days after the end of treatment. These data indicate that 7 days of dexamethasone therapy has significant but short-term effects on cortisol secretion and possibly on weight gain. PMID- 3050008 TI - Isolation and analysis of ketoconazole resistant mutants of Saccharomyces cerevisiae. AB - Nine mutants of Saccharomyces cerevisiae which are resistant to ketoconazole, have been isolated and characterized. In each case the mutation is nuclear in origin and allelic to a previously described mutation, erg3, which gives rise to a block in the delta 5-6 desaturation step of ergosterol biosynthesis. The significance of this second site mutation to the point of inhibitory action of ketoconazole, that is the P-450-mediated C-14 demethylation of lanosterol, is discussed. PMID- 3050009 TI - Morphological aspects of gastrointestinal tract invasion by Candida albicans in the infant mouse. AB - The infant mouse has proved to be a useful model for examination of various aspects of gastrointestinal and systemic candidosis. Oral-intragastric inoculation of 5-6-day-old mice with yeast of a virulent strain of Candida albicans (CA30) resulted in systemic spread within 30 min after challenge. Histological examinations of the gastrointestinal (GI) tract have shown that the highest frequency of invasion of the mucosa by yeast cells occurred in the region of the jejunum 1-3 h after inoculation. Results of ultrastructural examinations of sites where the fungus invaded the bowel wall suggested that C. albicans yeast cells are capable of progressive extracellular digestion of the intestinal mucus barrier and microvillus layer, followed by intracellular invasion of columnar epithelial cells. Minimal disruption of cytoplasmic contents of the host epithelial cells appears to result from invasion and transmigration of the pathogen. The infant mouse model is suggested to be well suited for localization of extracellular products of C. albicans yeast in vivo which may play pivotal roles in the invasion of host tissue during GI candidosis. PMID- 3050010 TI - Respiration of medically important Candida species and Saccharomyces cerevisiae in relation to glucose effect. AB - Strains of medically important Candida species (C. albicans, C. tropicalis, C. parapsilosis and C. [Torulopsis] glabrata) and Saccharomyces cerevisiae were examined for a glucose effect on respiratory activity. Reduced O2-consuming ability and a relative decrease in cytochrome type c, as determined by polarography and spectrophotometry, respectively, were observed in glucose-grown S. cerevisiae cells in contrast with acetate- or ethanol-grown cells. In glucose grown cells of C. glabrata, O2 consumption was also reduced without any change in the cytochrome pattern compared to acetate-grown cells, while no such decrease was detected in any of the other strains of Candida species tested. These results suggest that the medically important Candida species, except for C. glabrata, can be categorized as members of the glucose-insensitive yeast type with respect to respiration. PMID- 3050011 TI - Epidemiology of an outbreak of Candida endophthalmitis in heroin addicts: identification of possible source of infection by biotyping. AB - Biotyping was employed to investigate possible sources of Candida endophthalmitis in heroin addicts. Isolates of Candida albicans recovered from patients and from injection paraphernalia, including lemon juice diluent from lemon-shaped plastic containers, were biotyped. The predominant biotypes were 153, 15 3/7. Similar biotypes were recovered from lemon, mouthwash, mouth swab and vitreous samples. PMID- 3050012 TI - Intensive care course following liver transplantation in children. AB - We report the postoperative intensive care course of 16 children who underwent 18 orthotopic liver transplantation (OLT) procedures in London, Ontario and compare this experience in our developing transplant center with that reported from the Children's Hospital of Pittsburgh. Assisted ventilation was required in all children, with six requiring ventilation for greater than three days. Six children required positive end expiratory pressure (PEEP) therapy and hypertension was common. Physiologic stability index score was initially high in all patients, but fell on subsequent days. Intensive care survival was 100% with 69% long-term survival, which compared favorably with the information from Pittsburgh. Septic complications, despite immunosuppressive therapy were rare, but hypocalcemia and hypomagnesemia were common. PMID- 3050013 TI - Blunt pancreatic injuries in children: the role of percutaneous external drainage in the treatment of pancreatic pseudocysts. AB - During the past 10 years, 26 cases of blunt pancreatic trauma were diagnosed in our institution. In 42.3% (11/26) the accident was bicycle-related. Seventy-three percent of patients were seen within 48 hours of injury. The most frequent clinical presentations included abdominal pain, tenderness and vomiting. Diagnosis of pancreatic injury was suggested by hypermylasemia in most cases. Associated trauma was seen in seven patients (26.9%) and it was intraabdominal in four (15.3%). Computerized tomography (CT) scan is the single most useful radiologic investigation in evaluating pancreatic trauma. Ultrasound, although less accurate than CT scan in determining the severity of the initial injury, is useful in the evaluation and treatment of pancreatic pseudocysts. Pancreatic pseudocysts developed in ten patients. Spontaneous resolution occurred in five (50%). In three patients, percutaneous external drainage (PED) was successful in treating pancreatic pseudocysts without complications or recurrence at 11, 19, and 31 months. PED is a suitable form of treatment in selected cases of pancreatic pseudocysts. Results in children are better than in the adult population, probably due to the absence of primary pancreatic pathology. We believe that PED should be considered the primary therapeutic procedure for traumatic pancreatic collections prior to more invasive surgical treatment, when there is no evidence of pancreatic duct transection on CT scan. PMID- 3050014 TI - Simultaneous recording of fetal breathing movements and body movements in twin pregnancy. AB - Fetal breathing and body movements were simultaneously evaluated in twin pregnancies in order to determine to what extent these activities occur in a synchroneous pattern in both twin fetuses and if fetal position, presentation or sex have an influence on their behavior. Thirty healthy pairs of twins at 34-37 weeks of gestation were studied. Twenty-six percent of fetal body movements and 49% of breathing movements occurred simultaneously in both fetuses. The overall total simultaneous fetal activity rate was 53.3%. The length of breathing movements and total activity (summation of breathing and body movements) of the fetuses positioned on the right side of the uterus were significantly longer than in fetuses positioned on the left side of the uterus (p = 0.002) and (p less than 0.0001) respectively. This was also true for subgroups where only fetuses in the same presentation or of the same sex were compared. It is concluded that the fetus positioned on the right side of the uterus is more active and that fetal sex or presentation had no significant effect on intrauterine fetal activity in twin pregnancies. PMID- 3050015 TI - Intrauterine treatment of idiopathic hydrops fetalis. AB - Seven fetuses with idiopathic hydrops fetalis (IHF) were treated in utero by injecting albumin into the fetal abdominal cavity and by removal of accumulated fluid from the serous cavities. Signs of hydrops fetalis disappeared in utero in one, and skin edema significantly decreased in another. In the other five, signs of hydrops fetalis remained unchanged in utero. The hourly fetal urine production rate (HFUPR) increased after albumin injection in three of five. The interval between the initial diagnosis and delivery ranged from 3 to 14 weeks. Gestational age at the time of delivery ranged from 33 to 40 weeks. There were no stillbirths. Two of three without pleural effusion survived, but four with pleural effusion died of respiratory failure during the neonatal period due to pulmonary hypoplasia. These results indicate that albumin injection into the fetal abdomen in utero deserves further attention and that other therapeutic methods should be established to enhance the development of the lungs in cases of intrauterine treatment of IHF with pleural effusion. PMID- 3050017 TI - Renal transplantation during pregnancy--a case report. AB - A case of renal transplantation during the 12th week of pregnancy is presented. An episode of rejection 6 days after surgery was treated satisfactorily. At the 18th week, the patient showed a mild hypertension which was treated by hidralazine. At the 30th week a fetal pyelouretheral stenosis with celiciar dilatation was diagnosed and the mother had another rejection episode. At the 33rd week a cesarean section was performed after a pathological NST. Neonatal and maternal developmental courses were good. PMID- 3050016 TI - Iniencephaly: prenatal ultrasonographic diagnosis--a case report. AB - A case of iniencephaly, diagnosed prenatally at 19 weeks of gestation, is presented. The main findings of the ultrasound examination were anencephaly, almost complete absence of cervical structures and cervical and thoracic rachischisis. Amniotic fluid alpha-fetoprotein was markedly elevated. The diagnosis of iniencephaly was confirmed after termination of pregnancy. This case demonstrate the feasibility of the prenatal diagnosis of this rare neural tube defect. PMID- 3050018 TI - Microbiologic assessments to enhance periodontal diagnosis. AB - Associations between microbes and progressive periodontal lesions provide the biologic rationale for using specific organisms as bacteriologic markers for disease activity. Rapid, specific and relatively inexpensive laboratory tests to identify bacteria have facilitated increased testing by clinicians. However, assays must be interpreted with regard to current concepts of etiology and pathogenesis of periodontal diseases. This paper addresses commonly used microbiologic assays and their shortcomings. PMID- 3050019 TI - Enzyme activity in crevicular fluid for detection and prediction of clinical attachment loss in patients with chronic adult periodontitis. Six month results. AB - Previous reports have described a method by which multiple constituents can be analyzed from a sample of gingival crevicular fluid (GCF) collected with a precut filter paper strip. In this study the relationship of changes in GCF levels of the vertebrate (lysosomal) enzymes beta-glucuronidase (BG) and arylsulfatase (AS) and the cytoplasmic enzyme lactate dehydrogenase (LDH) was evaluated longitudinally in reference to loss of clinical attachment in patients with existing chronic adult periodontitis. Thirty-six patients were followed for six months. Clinical attachment loss was recorded as the change between the baseline and three month examinations, and the three- and six-month examinations. GCF analysis was performed at baseline and three months. Three groups of patients were identified based on disease progression. Group I patients (N = 5) displayed a generalized form of disease activity. In these patients we observed clinical attachment loss of at least 2.0 mm at a minimum of three unrelated sites. Group II patients (N = 4) displayed a localized form of disease activity. In these patients clinical attachment loss of at least 2.5 mm occurred at one site, or two anatomically related sites. Group III patients (N = 27) did not display clinical attachment loss as defined here. Enzyme analysis was evaluated as a whole mouth score (the per cent of samples from a patient in which enzyme activity was at least twice the population mean) and at individual samples. Group I patients could be identified by elevated whole mouth scores for BG, while Group II patients could not be identified by whole mouth scores for any of the enzymes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3050020 TI - Growth and migration of gingival epithelial cells on mineralized and partially demineralized root surfaces in an in vitro system. AB - The capacity of epithelial cells to migrate and grow from gingival explants cultured on mineralized and partially demineralized root surfaces in an in vitro system was assessed. Explants of attached gingiva were obtained from mongrel dogs, cut into rectangular pieces (1 x 2 mm) and cultured either on mineralized or partially demineralized cementum in a defined culture medium containing transferrin, insulin, epidermal growth factor, cortisone, high density lipoprotein and selenium. After seven days of culture, the specimens were prepared for scanning electron microscopic examination. The amount of epithelial outgrowth from each explant was assessed by measuring the distances between the four aspects of the rectangular explant and the furthest epithelial cell located opposite to each of these aspects. The mean value obtained for epithelial outgrowth of explants grown on mineralized cementum was six times higher than that for explants cultured on partially demineralized cementum. These results indicate that partially demineralized cementum does not support epithelial growth and migration in vitro. PMID- 3050021 TI - [Pyrogens and endotoxins. II. Properties and control of pyrogens]. PMID- 3050022 TI - [The immune system. General aspects]. PMID- 3050023 TI - Sex-related differences in hepatic drug-oxidizing capacity of streptozotocin induced diabetic rats. AB - Male and female animal models of diabetes were prepared by treating rats with streptozotocin (STZ). Trimethadione (TMO) metabolism was depressed in male but increased in female diabetic rats. The insulin treatment normalized rats of both sexes. Serum dimethadione/TMO ratios at 2 h correlated with the elevated blood glucose levels in male and female control, the STZ-induced diabetic and the insulin-treated diabetic rats. Treatment with STZ in male rats affected the metabolism of antipyrine, decreasing urinary excretion of norantipyrine (NORA) urine but increasing elimination of 4-hydroxyantipyrine (OHA). In female diabetic rats, the amounts of the three major metabolites, NORA, OHA and 3-hydroxymethyl-3 norantipyrine and the total (conjugate + free) were increased compared to the control. In the insulin-treated groups, these changes were normalized. In conclusion, our study showed that the effect of STZ-induced diabetes on drug metabolism varies with the sex and the drugs used. Insulin normalized all these diabetic changes. PMID- 3050024 TI - A review of the literature pertaining to the psychosocial responses of school aged children to hospitalization. PMID- 3050025 TI - Positive mood and helping behavior: a test of six hypotheses. AB - Past research has shown rather consistently that positive mood states lead to increased helpfulness. In an expanded analysis of the published literature, we examined six distinct views about this relation: the focus of attention, objective self-awareness, separate process, social outlook, mood maintenance, and concomitance hypotheses. For each of 61 positive affect conditions in which it was possible to generate an effect-size estimate corresponding to the relative degree of helpfulness exhibited by positive mood subjects (compared with neutral affect subjects), judges assessed the contextual levels of variables relevant to each of the six hypotheses by reading the Method section of each article. Higher order partial correlation coefficients were then calculated to isolate the independent contribution of each of the theoretically relevant variables to the variation among the 61 effect sizes. The results support the focus of attention, separate process, social outlook, and mood maintenance hypotheses, and partially support the objective self-awareness and concomitance hypotheses. PMID- 3050026 TI - The effects of some protein-modifying reagents on the interaction of colicins A, E2, E3, and K with their respective Escherichia coli cell receptors. AB - Colicins attach themselves--through specific protein-protein interactions--onto receptors in the outer membrane of sensitive bacterial cells. An attempt was made to analyze amino-acid groups and attractive forces involved in this interaction, following treatment of either colicins A, E2, E3 and K or sensitive bacteria with various physico-chemical factors and several protein-modifying reagents. The amounts of colicin bound were checked by a quantitative biological assay. Ionic conditions and specific spatial conformation of both colicin and its receptor molecules are crucial in their interaction and, hence, in the biological effect of colicin. Formaldehyde, naphthalene-diisocyanate and osmium tetroxide strongly inhibit the binding ability of all colicins tested. The results suggest that NH2 and SH groups are involved in their binding onto receptors; also, CH3S groups seem to be engaged in the attachment of colicins E2 and E3 and phenol-OH groups in that of colicin K. The possible involvement of further groups (NH, SH etc.) should be checked using more specific reagents. The attitude of colicins E2 and E3 to their common receptor Btu B protein is nearly, but not completely the same. Receptors for all colicins tested should be oxidized to achieve optimal interactions; obviously, carbonyl groups are produced and newly formed anions increase the negative load of bacterial surface. In agreement, reduction of at least colicins A and E3 enhances their receptor binding reactivity. The binding capacity of each receptor can be modulated by a set of amino acid reagents in a specific manner. PMID- 3050027 TI - Vagotomy by the local anesthetic procaine hydrochloride: a study in the rat. AB - Transmission of nerve impulses is prevented by the local anesthetic procaine hydrochloride. This study was undertaken in rat models, in which the vagus nerve mediates gastric acid secretion, to examine the action of 1 mL of 5% oral procaine on this secretion. The H+ output (mean +/- SEM) associated with pylorus ligation for 1 h (61 +/- 7 mumol) was depressed by vagotomy or procaine (4.7 +/- 0.5 mumol and 5.8 +/- 0.7 mumol, respectively; p less than 0.001). In the rat without pylorus ligation, reserpine (0.1 mg/kg, ip) stimulated H+ output .4 +/- 0.7 mumol; p less than 0.001). Vagotomy or procaine depressed .4 +/- 0.5 and 3 +/ 0.5 mumol, respectively versus 12.4 +/- 0.7 mumol han 0.001), and reserpine failed to influence this action. In the same in (1 unit/kg, ip) stimulated H+ output (36 +/- 3.1 versus 12 +/- 0.6 than 0.001), but had no effect on its depression by vagotomy (3 +/- /- 0.5 mumol) or procaine (4 +/- 0.4 versus 3.9 +/- 0.4 mumol). In the gastric diversion rat, a model for collecting gastric secretion from just distal to the pyloric sphincter into a plastic bag, the effect of vagotomy on the N+ output was similar to that of procaine (3.5 +/- 0.4 and 2.9 +/- 0.3 mumol, respectively, versus 14.8 +/- 0.5 mumol; mean +/- SEM.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3050028 TI - Plantar pyogenic granuloma. PMID- 3050030 TI - Author and subject indexes to Volumes 358-394 (January 1985-December 1987). PMID- 3050032 TI - Prosthodontic rehabilitation following total mandibular reconstruction. A clinical report. PMID- 3050031 TI - Single-villus analysis of disaccharidase expression by different regions of the mouse intestine. AB - 1. The present results describe how a new technique of whole-tissue cytochemistry can be combined with automatic scanning of microdissected villi to measure the capacity of individual villi to hydrolyse disaccharides in different parts of the small intestine. 2. Intact villi from the mouse proximal jejunum are found to be eight times more effective than ileal villi in hydrolysing 2-naphthyl-alpha-D glucoside, an artificial substrate for enzymes normally hydrolysing sucrose, maltose, isomaltose and trehalose in adult intestine. Homogenates of jejunal scrapings are four times more effective than ileal homogenates in hydrolysing this substrate. This discrepancy arises from relating enzyme activities to homogenate protein in cases where intestinal structure changes. 3. The eightfold difference in villus alpha-glucosidase activity is associated with a threefold difference in villus surface area. This discrepancy in turn reflects changes in the capacity of individual enterocytes to express alpha-glucosidase during migration along the crypt-villus axis. These results emphasize the futility of trying to gauge intestinal function from measurement of intestinal structure. 4. Differences between ileal and jejunal villus alpha-glucosidase activities have been further partitioned into those depending on villus structure and those depending on enterocyte development. Present results are discussed in relation to the ability of luminal nutrition to maintain a proximal-distal gradient of digestive enzyme function in the small intestine. The general applicability of this method of analysis to other studies of adaptive response is also emphasized. PMID- 3050033 TI - Occlusal adjustment for initial treatment and prevention of the cracked tooth syndrome. PMID- 3050035 TI - Film thickness measurements of a paint-on die spacer. PMID- 3050034 TI - The effect of enamel etchant on the solubility of three calcium hydroxide bases. PMID- 3050029 TI - Further characterization of the cardiovascular effects of the dopamine beta hydroxylase inhibitor SK&F 102698 in conscious hypertensive rats. AB - The dopamine beta-hydroxylase inhibitor SK&F 102698 was characterized by studying its cardiovascular effects in hypertensive rats. The antihypertensive effects of SK&F 102698 (50 mg/kg p.o.) were studied in three different rat models of hypertension. In spontaneously hypertensive and deoxycorticosterone acetate-salt rats SK&F 102698 produced blood pressure reductions of approximately 21 and 23%, respectively. In contrast, SK&F 102698 did not produce a significant decrease in blood pressure in 2-kidney, 1-clip Goldblatt hypertensive rats. The antihypertensive mechanism of action of dopamine beta-hydroxylase inhibition was probed with the selective DA1-receptor antagonist SCH 23390, which produced an attenuation of the antihypertensive effects of SK&F 102698. Experiments were designed to separate the peripheral from the central components of the cardiovascular effects of SK&F 102698. In spinal cord stimulated pithed spontaneously hypertensive rats, SK&F 102698 reduced blood pressure but not heart rate, indicating a peripherally mediated vasodilation and a centrally mediated heart rate effect. Furthermore, when SK&F 102698 was administered directly into the fourth ventricle of conscious spontaneously hypertensive rats, a pronounced bradycardia and lowering of blood pressure was observed. SCH 23390 (200 micrograms/kg i.v.) and l-sulpiride (1 mg/kg i.v.) inhibited the cardiovascular effects of SK&F 102698 administered into the fourth ventricle. These data indicate that inhibition of dopamine beta-hydroxylase with SK&F 102698 results in both peripherally and centrally mediated cardiovascular effects and suggest that central dopamine receptors contribute to the control of systemic blood pressure in hypertensive models associated with an increased sympathetic outflow. PMID- 3050036 TI - Bond strength of joined posterior light-cured composites: comparison of surface treatments. PMID- 3050037 TI - A two-stage impression technique for distal-extension removable partial dentures. AB - Partially edentulous mouths with distal-extension ridges present the challenge of correctly registering two tissues as dissimilar as teeth and edentulous ridges. A technique is described that makes it possible to make impressions with a combination of different impression materials within the same tray and have a firm support that assures even distribution of the impression material and faithfulness of reproduction. PMID- 3050038 TI - Occlusal considerations for partially or completely edentulous skeletal class II patients. Part I: Background information. AB - Although approximately 15% of the population may be classified as having the skeletal class II relationship, this group of patients is far from homogeneous. Two prototypes were used to delineate various problems in the prosthodontic occlusion that dentists may encounter with these patients. A satisfactory occlusion is difficult to achieve because of skeletal discrepancies, limited space for occlusal contact, steep guidance factors, and the necessity for multiple eccentric occlusal contacts because of the significant range of mandibular motion. PMID- 3050039 TI - Clinical and statistical analyses of human clinical trials with the single crystal aluminum oxide endosteal dental implant: five-year results. PMID- 3050040 TI - In vivo wear. Part II: Wear and abrasion of composite restorative materials. PMID- 3050041 TI - A cera-platin crown with an all-porcelain facial margin. PMID- 3050042 TI - Conversion of a conventional attachment to magnets. PMID- 3050043 TI - A technique for limiting reduction of overextended denture borders. PMID- 3050044 TI - Reversible hydrocolloid: the standard of excellence. AB - Reversible hydrocolloid is the oldest elastic dental impression material and, with the appropriate methodology, the advantages outweigh the disadvantages. Skill, care, and comprehension of the physical properties of the materials ensure success. PMID- 3050045 TI - Surface treatment of gold alloys for adhesion. PMID- 3050046 TI - Adhesive bonding of composites to a casting alloy. PMID- 3050048 TI - The effects of manipulative variables on the color of ceramic metal restorations. AB - Certain variables were tested for their influence on the color of ceramic metal restorations. The variables included firing temperature, condensation technique, modeling liquid, and brand of porcelain. In addition, the color of different nominal shades was compared. The color was measured with a Minolta CR-121 small area colorimeter. The small (7 mm2) measuring area enabled readings to be made on actual crown specimens rather than disks. The results were expressed in the CIELAB color system enabling comparisons to be made related to visual perception. The following conclusions can be drawn from the study. 1. The small-area colorimeter is able to detect statistically significant and perceivable color differences between shades of porcelain. The variation in color parameters associated with making replications of a restoration is significantly greater than the error associated with making repeated color measurements of the same restoration. 2. Restorations made with the different brands of porcelain studied have noticeably different colors despite having the same nominal shade. 3. Color changes caused by the choice of modeling liquid were not statistically significant in this study. If the mean color differences are truly representative, using Rainbow or Carv-eze modeling liquids with Vita VMK porcelain may produce color changes that are just barely detectable. 4. The manipulative variables of firing temperature and condensation have little influence on the color of the restorations. PMID- 3050049 TI - Occlusal considerations for partially or completely edentulous skeletal Class II patients. Part II: Treatment concepts. AB - Four basic concepts must be considered when developing the prosthetic occlusion for skeletal class II patients: (1) Centric relation must be used as a reference position to relate the mandible to the maxillae; (2) the posterior denture teeth must be positioned close to their former positions; (3) freedom of movement must be created in eccentric movements; and (4) multiple occlusal contacts must be provided in centric and eccentric positions. We have described a method that incorporates these basic concepts and provides a harmonious occlusal arrangement for both edentulous and partially edentulous class II patients. PMID- 3050047 TI - The sealing properties of temporary filling materials. PMID- 3050050 TI - A modified direct retainer design for distal-extension removable partial dentures. AB - The rationale for designing a direct retainer for a distal-extension removable partial denture is described. The advantages of an L-bar clasp arm over the I-bar clasp arm are discussed. The retentive surface of an abutment tooth is divided into three zones according to the clasp tip movement and the importance of placing the retentive tip in the zone of vertical movement is emphasized. PMID- 3050051 TI - An alternate centric relation recording technique for a distal-extension removable partial denture. PMID- 3050052 TI - Tissue-integrated implants for the partially edentulous patient. AB - Patients who have been dissatisfied with removable prostheses and have sufficient alveolar or cortical bone may be appropriate candidates for osseointegrated implants. This article demonstrates the clinical application of osseointegrated implants for the partially edentulous patient as a viable alternative treatment. PMID- 3050054 TI - Dimensional stability of injection and conventional processing of denture base acrylic resin. PMID- 3050055 TI - An alternative method of border molding. AB - An alternative method of border-molding using a syringe to place the impression compound is more efficient and allows border molding of an entire quadrant at a time. This method enables the dentist to deliver an even flow and control the amount of impression compound expressed in a neater and clearer manner. PMID- 3050053 TI - Prosthodontic rehabilitation of the partially edentulous trauma patient by using osseointegrated implants. AB - Osseointegrated dental implants provide a viable alternative of tooth replacement. Although certain patients may greatly benefit from this method of treatment, implants are not a panacea. Success is the culmination of good case selection, thoughtful treatment planning, meticulous clinical and technical attention to detail, and proper maintenance care. PMID- 3050056 TI - Is tooth wear a correlate of composite microwear? PMID- 3050057 TI - Penetration of etched enamel by bonding agents. PMID- 3050058 TI - Paraquat poisoning: biological presentation. PMID- 3050059 TI - Anxiety, panic and phobic disorders: an overview. AB - This paper reviews anxiety, panic, and phobic disorders as they were described in landmark works, along with more recent epidemiologic studies of the disorders. The author discusses clinical syndromes of anxiety as outlined in the DSM-III: agoraphobia, social phobia, generalized anxiety disorder, panic disorder, simple phobic states, and obsessive-compulsive disorder, relating them to Phobic Anxiety Depersonalization Syndrome and to earlier descriptions by Westphal and Benedict. The paper addresses the problem of delineating anxiety and phobic states from depressive disorders, with regard to diagnosis and treatment outcome. Various etiological bases of agoraphobia, panic, and anxiety disorders are suggested: heredity, life events and circumstances, family background and developmental history, the premorbid personality, and some psychological aspects. Several questions are explored on the relationships of agoraphobia, anxiety and panic attacks. For example, is agoraphobia a new disease or one stage in the development of severe chronic anxiety? Are the phobias of agoraphobia acquired by conditioning or learning? Are "panics" spontaneous or physiological? Are panic attacks the first event in the primary cause of agoraphobia? For future work the authors propose a reassessment of the prevalence of agoraphobia and related disorders, a more careful definition of the agoraphobic disorders, and thorough clinical investigation of the various treatment modalities in well-defined populations. The past twenty years' achievements in behavioural and pharmacological treatments for agoraphobia are briefly recapitulated. PMID- 3050060 TI - Tricyclic therapy of the DSM-III anxiety disorders: a review with implications for further research. AB - Existing data suggests dramatic efficacy for all tested tricyclics in disorders involving spontaneous panic attacks; moderate efficacy for clomipramine but not other tricyclics in obsessive-compulsive patients; possible, but as yet not well established, tricyclic efficacy in generalized anxiety; and lack of efficacy in simple phobias. Further studies in all DSM-III Anxiety Disorders are both warranted and required to answer the nosological, pathophysiological and treatment questions raised by these findings. PMID- 3050061 TI - Monoamine oxidase inhibitors in anxiety disorders. AB - Monoamine oxidase inhibitors (MAOI's) have been shown to be significantly superior to placebo in the treatment of some anxiety disorders, particularly agoraphobia and mixed anxiety--depressive states. There is no convincing evidence that MAOI's are effective treatment in pure anxiety states, whether or not panic is present as a major symptom, although they are effective in so-called endogenous anxiety. Many past published studies of MAOI's have yielded poor results because the drugs have been prescribed for insufficient time (less than four weeks) or at too low dosage. There are no important therapeutic differences between the MAOI's apart from the faster speed of response with the nonhydrazine compound, tranylcypromine. Treatment often has to be long-term, and some degree of pharmacological dependence may develop. A few clinical studies have compared the efficacy of MAOI's and tricyclic antidepressants in anxious disorders. There is growing evidence that MAOI's are somewhat more effective than tricyclic antidepressants in the treatment of anxiety disorders and when phobic anxiety is an important component of a depressive disorder. PMID- 3050062 TI - The epidemiology of anxiety disorders: rates, risks and familial patterns. AB - This paper reviews what we know about the epidemiology and familial patterns of anxiety disorders. Focus is on the current studies based on specified diagnostic criteria. Data are presented, when available, on the subclassifications of the anxiety disorders. Data from epidemiologic and family studies support the notion that anxiety disorders have a relatively high prevalence and are familial, that they are heterogeneous, and that some are related to depression. It suggests that there is an increased probability that a person with one anxiety disorder will have another or will have a major depression during his or her lifetime. Data also suggest that panic disorder has the most severe consequence in terms of morbid risk to first-degree relatives, particularly risk to children, and that there may be a relationship between adult and childhood anxiety disorders. Potential research areas are given. PMID- 3050064 TI - Battered women and abusive partners: treatment issues and strategies. PMID- 3050063 TI - Kinesthetic aftereffect and augmenting/reducing: a two-session procedure, and hence identification of stimulus-governed subjects, is contraindicated. AB - A few of the recent researchers of kinesthetic aftereffect (KAE) as an index of augmenting/reducing have continued to employ a two-session procedure. Findings that have accrued in the past decade indicate (a) The first administration of KAE is a reliable (internally consistent) and valid index of augmenting-reducing; (b) there are carry-over effects from the first to the second administration that bias the second session's preinduction scores; (c) KAE scores from sessions after the first do not relate to first-session scores (low retest reliability) and do not measure augmenting/reducing; and (d) unless special procedures are undertaken to avoid using the biased second- (or later-) session preinduction scores, a KAE procedure involving more than one session is contraindicated. When we reached a similar conclusion earlier (Baker et al., 1974), Petrie (1974) disagreed, arguing that a two-session procedure was needed to identify and eliminate an atypical subgroup, the "stimulus governed." The case for determining which subjects are stimulus-governed is assessed and found wanting. Except in special circumstances, a one-session KAE procedure, in which all preinduction trials precede the first exposure to aftereffect induction, is indicated. PMID- 3050065 TI - Angiologic observations following autologous vein grafting and free radial artery flap elevation. AB - Fifteen cases with radial forearm flap harvesting and autologous vein-graft reconstruction of the missing radial artery portion, are reported. Post-repair follow-up examinations, using segment plethysmography, photoplethysmography, and Doppler ultrasound, demonstrated an angiologic donor site morbidity, even when radial artery reconstruction was performed. Typically diminished blood pressure occurred, in comparison with the contralateral healthy extremity. Index shifting of pulse wave peaks, as well as widening of pulse wave bases occurred, especially in the thumb and index finger. These latter findings appear to be discrete indicators of arterial insufficiency. PMID- 3050066 TI - Vessel anastomosis using a venous cuff and two sutures: an experimental study in rat femoral and epigastric vessels. AB - Using rat femoral and epigastric vessels, the efficacy of an autogenous venous cuff and reduction in the number of required sutures were examined. Cut vessels were anastomosed with two sutures using 11-0 nylon, and the anastomotic site was covered with a venous cuff. After two weeks, the patency of the anastomosed vessels was evaluated. Although results in large-caliber femoral veins were poor, the femoral arteries and epigastric vessels demonstrated high patency rates. In microscopic and scanning electron microscopic evaluations, gaps and depressions between sutures were observed, but they were smoothly covered by endothelium. These results suggest the usefulness of the venous cuff in microvascular surgery, especially in anastomosing fine-caliber vessels. PMID- 3050067 TI - Delayed hypersensitivity and lesions following isoimmunization with modified rat male accessory glands: kinetics of induction. AB - The kinetics of the cellular immune response to rat male accessory glands were studied in Wistar rats isoimmunized with modified rat male accessory glands extract and complete Freund's adjuvant at 0, 30 and 45 days. The animals were divided into seven groups, and each group was sacrificed weekly. One immunization was sufficient for the induction of 2-, 6- and 24-h footpad reactivity. The reaction increased until 21 days post-immunization. After the second injection the reaction decreased and was negative 12 days later. Migration inhibitory factor (MIF) activity monitored by a mixed-direct assay was demonstrated in rats from all groups except in the animals studied at day 42 in which macrophage migration was markedly stimulated. The absence of MIF activity correlated with a lack of delayed type hypersensitivity (DTH) response. The humoral response was studied and detected by passive hemagglutination in a few sera after the first immunization. A second injection was necessary to obtain a more frequent occurrence and higher titres of antibodies. Histological modifications in the target organs started to appear in the group of animals studied at 35 days and were characterized by a mononuclear infiltrate in the prostate, coagulating glands and seminal vesicles. In several cases there was also infiltration of polymorphonuclear cells. Specimens obtained at 35 days showed the most severe lesions. PMID- 3050069 TI - A synthetic luteinizing hormone releasing hormone vaccine. I. Conjugation and specificity trials in BALB/c mice. AB - The immunobiology of luteinizing hormone releasing hormone (LHRH) was explored, to provide a conceptual and practical basis for the use of LHRH in immunocastration. Cysteine substituted analogues of LHRH were synthesized including Cys1-LHRH (C1-LHRH), Cys6-LHRH (C6-LHRH) and Cys10-LHRH (C10-LHRH). These were reacted to carrier molecules using the heterobifunctional cross linking reagent m-maleimidobenzoylsulfosuccinimide ester (SMBS), producing peptide-carrier conjugates of known peptide content and conjugation orientation. This reaction regime was found to be rapid, efficient and allowed for easy control of peptide to carrier ratios. Conjugates were used in active immunization trials in BALB/c mice to characterize the murine immune response against LHRH. BALB/c mice were shown to have the capacity to recognise all three cysteine substituted LHRH analogues and to produce antibodies cross-reactive with native LHRH. The specificity of LHRH antisera generated was found to be dependent on the site of conjugation of the peptide to carrier molecule. C1-LHRH generated carboxy terminal directed antibodies, C10-LHRH generated amino terminal directed antibodies, while C6-LHRH could generate amino terminal directed or carboxy terminal directed antibodies, or both within a given animal. No intrinsically immunodominant epitopes were seen within the LHRH molecule. PMID- 3050068 TI - Topographical expression of class I major histocompatibility complex antigens on human amniotic epithelium. AB - The expression of class I major histocompatibility complex (MHC) antigens on different regions of human amnion was studied by the avidin-biotin-complex immunoperoxidase (ABC) technique using monoclonal antibodies. In contrast to previous reports, the use of higher affinity monoclonal antibodies and the sensitive immunoperoxidase method has allowed the identification of class I MHC antigens on the amniotic epithelium. The level of expression is different between cells from various parts of the amnion, with the amniotic epithelium from the edge of the placenta consistently showing the strongest reactivity. Some of the class I MHC antigens expressed by the amnion are similar to those expressed by extravillous trophoblast cells in that both show much weaker or no reactivity with the monoclonal antibody 61D2. PMID- 3050070 TI - Reliability of ultrasound in predicting uterine leiomyoma volume. AB - Recently ultrasound has been used to size and track individual myoma volumes for patients undergoing medical therapy. However, little is known about the specific performance characteristics and limitations of this technique with respect to volume measurements. We performed and prospectively interpreted serial ultrasound examinations on myoma patients and confirmed the location, size and number of myomas in the surgical specimens. The smallest detectable tumor was 2.7 cm in diameter. The specificity was 94%; reproducibility had a 14% coefficient of variation for tumors with diameters greater than 6 cm. We conclude that ultrasound is suitable for imaging and sizing myomas provided that the diagnosis is otherwise certain and the tumors tracked are large. PMID- 3050072 TI - Concurrence of ovarian cancer and dermatomyositis. A report of two cases and literature review. AB - The concurrence of dermatomyositis and ovarian carcinoma deserves special attention owing to the gravity of its prognosis. Until now, only scattered case reports on women suffering from this disease combination had been published. In this paper the information available in the literature on 28 women afflicted with these diseases is collated. In addition, two such patients treated in our hospital are described in detail. The review highlights the various aspects of this disease combination, emphasizing the quadrupled incidence, advanced stage and uniform epithelial origin of the malignancy as compared with that in the general female population with ovarian carcinoma. The value of a meticulous gynecologic evaluation was proven in one of our patients with dermatomyositis; in her, ovarian carcinoma was detected in its early phase. PMID- 3050071 TI - Transient failure of central opioid tonus and premenstrual symptoms. AB - In order to evaluate the relationships between endogenous opioid activity and premenstrual complaints, we subjected three groups of patients in the mid (days 8 12 prior to menses) and late (days 1-5 prior to menses) luteal phases of the cycle to a naloxone test and some of the patients to a luteinizing-hormone releasing hormone (LHRH) test. The premenstrual syndrome (PMS) group was composed of nine patients complaining of dizziness, irritability and depression close to menses for at least three years. The menstrually related migraine (MM) group was composed of 15 patients complaining of premenstrually related migraine. The common migraine (CM) group was made up of 16 women suffering from common migraine for years whose attacks occurred independently of menstrual cycle events. A group of seven fertile women served as controls. Every two days the patients filled out the Menstrual Distress Questionnaire for evaluation of their complaints. After the evaluation of spontaneous LH pulsatility for one hour, 4 mg of naloxone was injected as a bolus, and samples were collected every 15 minutes for 2 hours. Both estradiol (E2) and progesterone (P) were measured in basal samples from each naloxone test. LH responsiveness to LHRH was similar in the mid and late luteal phases and did not change between groups. In the mid luteal phase the LH response to naloxone in PMS and MM patients was similar to that in normal subjects, while CM patients had impaired LH secretion. In the premenstrual phase only the controls maintained an LH responsiveness similar to that observed in the mid luteal phase, while both PMS and MM lost the naloxone-induced LH release.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3050073 TI - Clinical characteristics of women with chlamydial cervicitis. AB - Four hundred ninety-three women were examined, had a cervical culture done for Chlamydia trachomatis and completed a questionnaire. Sixty-one, or 12.4%, were positive for Chlamydia on culture. Infected women were more likely to be younger, to be unmarried, to have had more sexual partners, to have had past abnormal cytology, to have spotting or postcoital bleeding, to have a mucoid or purulent cervical discharge and to have signs of cervical inflammation, especially friability. No characteristic finding in the past history, symptoms or physical examination combined sufficient sensitivity and specificity to serve by itself as a basis for selective testing for C trachomatis. It appears that multiple characteristics--including history, symptoms, physical examination and cytologic results--must be utilized to select patients for testing. PMID- 3050074 TI - Subsets of vulvodynia. AB - The frustrated patient with vulvar "burning" seldom has grossly abnormal physical findings, and the clinical significance and prognosis of vulvodynia have eluded both gynecologists and dermatologists. Evaluation of the physical findings on initial and follow-up visits of such patients during various treatment protocols has revealed distinct subsets of vulvodynia. In one series of 52 patients at the Emory Clinic, the following subsets were identified in decreasing order of frequency: (1) vulvar dermatoses, (2) cyclic candidiasis, (3) squamous papillomatosis, (4) vulvar vestibulitis, and (5) essential vulvodynia. These subsets may occur alone, simultaneously or sequentially; treatment for one condition may affect the onset of another. Vulvodynia may have multiple causes; use of the term for a patient's problem should prompt a thorough diagnostic evaluation. Although the subsets are not entirely exclusive of one another, each is identified by an improvement with specific therapeutic modalities. The recognition of multiple factors in vulvodynia is important to appropriate patient evaluation and management. PMID- 3050075 TI - Unilateral tubal twin pregnancy. A report of two cases. AB - Unilateral tubal twin pregnancy is exceedingly rare: only 91 cases have been reported in the English literature to date. Two additional such cases are described. PMID- 3050076 TI - Primary hyperparathyroidism in pregnancy. A report of two cases. AB - Primary hyperparathyroidism in pregnancy is an uncommon event, with approximately 100 reported cases in the literature. Two cases illustrate the common problems in diagnosing this disease in pregnancy and indicate that surgery should be considered the primary modality of treatment. These two are the 24th and 25th cases of surgery for primary hyperparathyroidism in pregnancy reported in the literature. PMID- 3050077 TI - Fetal umbilical velocimetry for the surveillance of pregnancies complicated by placenta previa. AB - Continuous-wave Doppler studies of the umbilical arteries were performed on 100 women with placenta previa prior to delivery in the third trimester and 100 control subjects matched for gestational age and with normally implanted placentas. The ratio of the maximum (systole [S]) and minimum (diastole [D]) velocities appeared to reflect downstream vascular resistance. An S:D ratio of greater than or equal to 3.0 indicated elevation. There was a statistically higher S:D ratio in the placenta previa group as compared to the control group, and that difference persisted even when growth-retarded fetuses were excluded from both groups. Placenta previa patients with S:D ratios greater than or equal to 3.0 were associated with a statistically higher incidence of adverse pregnancy outcomes. The sensitivity, specificity, and positive and negative predictive value of an elevated S:D ratio in predicting adverse pregnancy outcomes were 90.0%, 93.8%, 62.0% and 98.5%, respectively. Our data suggest that (1) placenta previa patients are at increased risk of intrauterine growth retardation and adverse pregnancy outcomes, (2) placenta previa patients with normal S:D ratios are at no greater risk of adverse outcomes than are controls, (3) when the S:D ratio is elevated in placenta previa patients, it is more likely to be associated with adverse pregnancy outcomes, and (4) placenta previa patients without growth failure have a higher S:D ratio than do those with normally implanted placentas. Umbilical artery velocimetry appears to be a useful adjunct to ultrasound in the surveillance of pregnancies complicated by placenta previa. PMID- 3050079 TI - First-trimester diagnosis of fetal abnormalities. A report of three cases. AB - First-trimester sonography is used extensively for dating pregnancy and in conjunction with chorionic villus sampling. It is crucial to examine the fetus carefully, even at this early stage, since some fetal abnormalities are detectable in the first trimester. In three cases encephalocele, thanatophoric dwarf and cystic hygroma were diagnosed with sonography in the first trimester. PMID- 3050078 TI - Postpartum blood flow velocity waveforms of the uterine arteries. AB - Uterine arterial velocity waveforms, recorded by pulsed Doppler technology, demonstrated a significant increase in vascular resistance after the first postpartum day until the fourth to sixth postpartum week. In patients with an isthmic postcesarean hemorrhage, partial placental retention and puerperal endometritis, the corresponding waveforms presented higher diastolic velocity components than was the case for normal puerperal courses, indicating that lower uterine vascular resistance might be associated with these complications. PMID- 3050080 TI - Royal National Hospital for Rheumatic Diseases--Bath. A 250th birthday party. PMID- 3050081 TI - Multicenter evaluation of synovectomy in the treatment of rheumatoid arthritis. Report of results at the end of five years. AB - A controlled multicenter evaluation of open surgical synovectomy in the treatment of rheumatoid arthritis (RA) was carried out over a period of 5 years after the operation. There was evidence of benefit in some knees but none in finger joints. Synovectomy performed at a stage of minimal damage did not result in fewer recurrences of disease activity than when done after greater damage had occurred. The results showed little longterm value of surgical synovectomy in the general treatment of RA or in the prevention of recurrences of disease activity or progressive articular damage. These results emphasize the need for controls in trials of this type. PMID- 3050083 TI - Phospholipase A2 activity associated with synovial fluid cells. AB - High activity of phospholipase A2 (PLA2) has been detected in synovial fluids (SF) in inflammatory arthritides. Since the source(s) of SF PLA2 has not been identified, we tested PLA2 content in SF cells obtained from 11 SF. Cell sonicates were prepared at 5 X 10(6) cells/ml. In the supernatants of the sonicated SF cells (n = 11), PLA2 activity ranged from 38-755 U/ml, mean 368 +/- 243 (SD) U/ml, compared to 5-64 U/ml, mean 31 +/- 15 (SD) U/ml in sonicates of normal peripheral blood PMN (n = 5) (p less than 0.0001). Spontaneous release of PLA2 from unstimulated SF cells ranged from 26-365 U/ml, mean 131 +/- 144 (SD) U/ml (n = 5), whereas spontaneous release from peripheral blood PMN was negligible. Neither 10(-8) M FMLP nor 5 X 10(-6) M dexamethasone altered extracellular PLA2 release. To assess whether PLA2 adsorbs passively to cell membranes through hydrophobic interaction, normal peripheral PMN were incubated in crude SF (n = 7) with PLA2 ranging from 4,000-24,300 U/ml, or with purified human SF PLA2 or Naja naja PLA2. We found that soluble PLA2 adsorbed to PMN membranes in a concentration dependent fashion. PLA2 activity in sonicates of PMN incubated in crude SF ranged from 185-358 U/ml compared to controls of 21-64 U/ml. Sonicates of PMN incubated with purified human SF PLA2 (5,000-30,000 U/ml) showed progressive concentration dependent increase in PLA2 from 7 +/- 4 to 212 +/- 11 (SD) U/ml (p less than 0.001). PMN incubated with Naja naja PLA2 also showed marked increase in the content of PLA2.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3050082 TI - IgG subclass distribution of antinative type II collagen and antidenatured type II collagen antibodies in patients with rheumatoid arthritis. AB - In 81 patients with rheumatoid arthritis (RA) with antibodies to native type II collagen, the frequencies of each IgG subclass to native type II collagen were IgG1 (70%), IgG2 (12%), IgG3 (84%) and IgG4 (6%) and to denatured type II collagen were IgG1 (86%), IgG2 (23%), IgG3 (86%) and IgG4 (6%). Thus serum antibodies to type II collagen in patients with RA were predominantly of the complement fixing subclasses IgG1 and IgG3. PMID- 3050084 TI - Primary juvenile Sjogren's syndrome. AB - Primary Sjogren's syndrome has rarely been reported in children. We report 3 patients who developed lacrimal and salivary gland involvement at ages 3, 7 and 9, respectively. The one who began at age 3 is the youngest we could find reported. Sjogren's syndrome-type involvement was documented objectively at both sites and associated connective tissue diseases were ruled out in all 3 patients. PMID- 3050085 TI - Radiologic vignette: primary disorders of articular cartilage in childhood. AB - We discuss several less well recognized disorders of articular cartilage which may affect pediatric patients, including relapsing polychondritis, idiopathic chondrolysis of the hip and chondrolysis after slipped capital femoral epiphysis. Idiopathic osteolysis is not considered in our review because it has been well described previously. PMID- 3050086 TI - Clinical trials with biological response modifiers in rheumatic diseases. PMID- 3050087 TI - Concurrent use of folinic acid and methotrexate in rheumatoid arthritis. AB - In a double blind placebo controlled crossover study, each arm of 4 weeks' duration, 20 mg of folinic acid/week or placebo were administered to 13 patients with rheumatoid arthritis. These individuals were receiving weekly intramuscular methotrexate (MTX) but were about to discontinue because of side effects. While there was considerable improvement in nausea during the study, the effect of folinic acid could not be differentiated from that of placebo. There was no adverse effect on control of disease activity. It therefore seems likely that polyglutamated tissue stores of MTX do not contribute to drug efficacy and in this format folinic acid could not be shown to be more useful than placebo in reducing drug induced nausea. PMID- 3050089 TI - Synthesis and pharmacological characterization of 1-phenyl-, 4-phenyl-, and 1 benzyl-1,2,3,4-tetrahydroisoquinolines as dopamine receptor ligands. AB - A series of 1-phenyl-, 4-phenyl-, and 1-benzyl-1,2,3,4-tetrahydroisoquinolines have been prepared as ring-contracted analogues of the prototypical D1 dopamine receptor antagonist SCH23390 [(R)-(+)-7-chloro-8-hydroxy-3-methyl-1-phenyl 2,3,4,5-tetrahydro-1H- 3-benzazepine]. The affinity and selectivity of these isoquinolines for D1 receptors was determined by three biochemical endpoints in membrane homogenates prepared from rat corpus striatum: the potency to complete for [3H]SCH23390 binding sites; the potency to compete for [3H]spiperone (a D2 receptor ligand) binding sites; and effects on dopamine-stimulated adenylate cyclase. Competitive binding measurements at D1 sites showed SCH23390 to possess the highest affinity, followed by 1-phenyl greater than 1-benzyl greater than 4 phenyl for the isoquinolines. These results were highly correlated with the ability of the test compounds to antagonize dopamine-stimulated adenylate cyclase (r = 0.98). None of the compounds alone stimulated cAMP formation at concentrations of 10 nM to 100 microM. D2 competition binding showed the 1-benzyl derivative to possess the highest affinity, followed by 4-phenyl greater than SCH23390 greater than 1-phenyl. The tertiary 1-phenyl derivative was more potent than the secondary 1-phenyl analogue in all assays. Interestingly, resolution and single-crystal X-ray analysis of the tertiary N-methyl-1 phenyltetrahydroisoquinoline showed the most active enantiomer to possess the S absolute configuration, in contrast to the benzazepine (R)-SCH23390. PMID- 3050088 TI - Renin inhibitors containing hydrophilic groups. Tetrapeptides with enhanced aqueous solubility and nanomolar potency. AB - Nineteen tetrapeptides containing statine (Sta) and 4-amino-5-cyclohexyl-3 hydroxypentanoic acid (ACHPA) were prepared. Solubility measurements of these compounds were carried out in H2O and in pH 7.4 phosphate buffer solution, and their partition coefficients were determined in a 1:1 1-octanol/sodium phosphate citric acid buffer system. The tetrapeptides were tested in vitro for their ability to inhibit porcine, canine, and human plasma renins. Four compounds, 6, 12, 14, and 20, were potent inhibitors against all renins tested (IC50 = 10(-9) M). Compound 12 was administered orally to dogs and substantially inhibited plasma renin activity for up to 5 h. The addition of polar groups to the C terminus of Sta- and ACHPA-containing tetrapeptides renders them soluble in aqueous milieu and provides a valuable tool with which to examine the role of the renin-angiotensin system in physiological and pathological circumstances. PMID- 3050090 TI - Oral absorption of cephalosporin antibiotics. 3. Synthesis and biological properties of 7 alpha-methoxy-7 beta-(arylacetamido)-3-chloro-3-cephem-4- carboxylic acids. AB - A series of 7 alpha-methoxy-7 beta-amido-3-chloro-3-cephem-4-carboxylic acids was prepared and evaluated for biological activity. When compared with the parent 7 non-methoxy analogues, these new 7 alpha-methoxy-3-chloro cephalosporins displayed diminished antimicrobial activity. PMID- 3050091 TI - The value of the study of natural history in genetic disorders and congenital anomaly syndromes. AB - The study of the natural history of genetic disorders and syndromes with congenital anomalies and dysmorphic features is a challenging and often neglected area. There are many reasons to pursue this type of research but it requires special clinical skills and a considerable amount of hard work. Setting up protocols and collecting data is complex and time consuming. Frequently, helpful clues for a particular disorder come from the study of the natural history of other disorders. Older affected subjects and unique cases with unusual features are often most important in unravelling the 'normal' course of a disease or recognising the basic defect. The study of natural history from individual patients and their records is complementary to population or registry based studies because it identifies individual variations and clinical heterogeneity. The understanding of the natural history of a particular disorder is of importance both to the affected person and their family and to the physicians caring for them. It is also useful to the basic researcher trying to determine the pathogenetic mechanism causing the disorder. In many ways, clinical geneticists have learned the art of caring for patients, as well as the challenges of clinical genetics, by becoming apprentices to and studying in depth specific disease entities. PMID- 3050092 TI - Use of computers in dysmorphology. AB - As a consequence of the increasing power and decreasing cost of digital computers, dysmorphologists have begun to explore a wide variety of computerised applications in clinical genetics. Of considerable interest are developments in the areas of syndrome databases, expert systems, literature searches, image processing, and pattern recognition. Each of these areas is reviewed from the perspective of the underlying computer principles, existing applications, and the potential for future developments. Particular emphasis is placed on the analysis of the tasks performed by the dysmorphologist and the design of appropriate tools to facilitate these tasks. In this context the computer and associated software are considered paradigmatically as tools for the dysmorphologist and should be designed accordingly. Continuing improvements in the ability of computers to manipulate vast amounts of data rapidly makes the development of increasingly powerful tools for the dysmorphologist highly probable. PMID- 3050093 TI - Microdeletion syndromes, balanced translocations, and gene mapping. AB - High resolution prometaphase chromosome banding has allowed the detection of discrete chromosome aberrations which escaped earlier metaphase examinations. Consistent tiny deletions have been detected in some well established malformation syndromes: an interstitial deletion in 15q11/12 in the majority of patients with the Prader-Willi syndrome and in a minority of patients with the Angelman (happy puppet) syndrome; a terminal deletion of 17p13.3 in most patients examined with the Miller-Dieker syndrome; an interstitial deletion of 8q23.3/24.1 in a large majority of patients with the Giedion-Langer syndrome; an interstitial deletion of 11p13 in virtually all patients with the WAGR (Wilms' tumour-aniridia gonadoblastoma-retardation) syndrome; and an interstitial deletion in 22q11 in about one third of patients with the DiGeorge sequence. In addition, a combination of chromosome prometaphase banding and DNA marker studies has allowed the localisation of the genes for retinoblastoma and for Wilms' tumour and the clarification of both the autosomal recessive nature of the mutation and the possible somatic mutations by which the normal allele can be lost in retina and kidney cells. After a number of X linked genes had been mapped, discrete deletions in the X chromosome were detected by prometaphase banding with specific attention paid to the sites of the gene(s) in males who had from one to up to four different X linked disorders plus mental retardation. Furthermore, the detection of balanced translocations in probands with disorders caused by autosomal dominant or X linked genes has allowed a better insight into the localisation of these genes. In some females with X linked disorders, balanced X; autosomal translocations have allowed the localisation of X linked genes at the breakpoint on the X chromosome. Balanced autosome; autosome translocations segregating with autosomal dominant conditions have provided some clues to the gene location of these conditions. In two conditions, Greig cephalopolysyndactyly and dominant aniridia, two translocation families with one common breakpoint have allowed quite a confident location of the genes at the common breakpoint at 7p13 and 11p13, respectively. PMID- 3050095 TI - Malformation syndromes: a review of mouse/human homology. AB - The purpose of this paper is to review the known and possible homologies between mouse and human multiple congenital anomaly syndromes. By identifying single gene defects causing similar developmental abnormalities in mouse and man, comparative gene mapping can be carried out, and if the loci in mouse and man are situated in homologous chromosome segments, further molecular studies can be performed to show that the loci are identical. This paper puts forward tentative homologies in the hope that some will be investigated and shown to be true homologies at the molecular level, thus providing mouse models for complex developmental syndromes. The mouse malformation syndromes are reviewed according to their major gene effects. X linked syndromes are reviewed separately because of the greater ease of establishing homology for these conditions. PMID- 3050096 TI - Are 'upper' and 'lower' neural tube defects aetiologically different? PMID- 3050094 TI - Dysmorphic syndromes with demonstrable biochemical abnormalities. AB - Many inborn errors of metabolism are associated with dysmorphic manifestations. In this review, we have attempted to correlate the dysmorphic features with the underlying metabolic defect or its consequences. Most of the defects which we have discussed affect the synthesis or degradation of macromolecules (for example, collagen, elastin, bone mineral, proteoglycans, glycoproteins, and triglycerides). Such defects may affect either a single enzyme or multiple enzymes in specific organelles, such as lysosomes or peroxisomes, or they may affect hormonal control of synthesis and degradation. Examples are also included of defects affecting the catabolism of simple molecules when accumulating metabolites have a secondary effect on macromolecules, as in homocystinuria. In a number of instances, however, the correlation between the biochemical abnormality and the dysmorphic features are not understood. Ultimately, all dysmorphic syndromes will be attributable to a biochemical defect or its effects. The aim of this overview is to provide an insight into the relationship between the two at the present time. PMID- 3050097 TI - Triphalangeal thumb. AB - Triphalangeal thumb (TPT), a rare malformation of uncertain pathogenesis, may occur as an isolated defect, in association with other malformations of the hands, or as a feature of a syndrome or sequence. Isolated TPT occurs in two functional types: opposable and non-opposable. The latter appears to be inherited as a simple autosomal dominant trait, while the former is generally sporadic. TPT is associated with a number of specific malformations of the hand or foot, several of which have a well documented autosomal dominant pattern of inheritance. TPT is a feature of a number of specific syndromes. In this setting it may be associated with radial hypoplasia, bone marrow dysfunction, congenital heart disease, lung hypoplasia or agenesis, anorectal malformations, sensorineural hearing loss, onychodystrophy, mental retardation, and other disorders. TPT serves as a useful marker in such patients; in conjunction with the clinical and radiological findings, it can help to establish the correct diagnosis, leading to appropriate management and genetic counselling. PMID- 3050099 TI - The telecanthus-hypospadias syndrome. AB - The telecanthus-hypospadias (BBB) syndrome is characterised by widely spaced inner ocular canthi and hypospadias of variable degree. Heterozygous females have telecanthus. We have summarised the historical and phenotypic findings of 21 patients in seven previous publications. We have also had the opportunity to evaluate personally 12 families with a total of 18 affected males. The most frequent anomalies in patients previously reported are telecanthus 21/21, hypospadias 19/21, cleft lip/palate or uvula 7/21, high, broad nasal bridge 15/15, cranial abnormality 6/21, congenital heart defect 5/21, cryptorchidism 9/21, and mental retardation 11/17. In our series, the most frequent anomalies include telecanthus 18/18, hypospadias 18/18, cleft lip/palate or uvula 8/18, high, broad nasal bridge 10/11, cranial abnormality 12/18, congenital heart defect 3/18, upper urinary tract anomaly 4/9, and mental retardation 10/12. There is also an increased incidence of like-sex twinning, 11/18 in our families. This syndrome must be more common than reflected in published reports. Based upon the observation that males are much more severely affected than females and the lack of male to male transmission, it appears that this condition is most likely to be inherited in an X linked fashion. Further elucidation of the phenotype and documentation of the inheritance is needed. The distinction between the telecanthus-hypospadias syndrome and the G syndrome also needs further clarification. PMID- 3050100 TI - Russell-Silver syndrome. PMID- 3050098 TI - Mitochondrial myopathy: a genetic study of 71 cases. AB - Of 71 index cases with histologically defined mitochondrial myopathy, 13 (18%) had relatives who were definitely affected with a similar disorder. Eight familial cases from four families were confined to a single generation. In five families maternal transmission to offspring occurred. There were no instances of paternal transmission, but one patient had an affected cousin in the paternal line. No consistent clinical syndrome or pattern of inheritance emerged for any identified defect of the mitochondrial respiratory chain, localised biochemically in 41 cases. Overall, the recurrence rate was 3% for sibs and 5.5% for offspring of index cases. Review of published reports of familial cases of mitochondrial myopathy suggests that the ratio of maternal to paternal transmission is about 9:1. We conclude that these disorders may be caused by mutations of either nuclear or mitochondrial genes. PMID- 3050101 TI - Pierre Louis Moreau de Maupertuis (1698-1759). AB - Maupertuis was one of the most important scientists and original thinkers of the 18th century. He refuted the preformationist theories of the time, and from his study of the inheritance of genetic traits proposed various ideas which forecast the genetic theory of inheritance. He applied the concept of probability to genetic problems and introduced experimental breeding as a means of studying the inheritance of genetic traits in animals. However, he considered his greatest contribution to science to be the 'Principle of Least Action'. Yet paradoxically it was this, through the vitriolic attacks by Voltaire which it engendered, which resulted in his work being largely ignored until recent times. PMID- 3050102 TI - A literature review on the efficacy of video in patient education. AB - Despite the recent dramatic increase in the use of video for patient education, there has been no critical assessment of this medium. In this paper, the author reviews 25 methodologically-sound studies in order to define the efficacy and limitations of video. Video is as good as and often more effective than traditional methods of patient education in increasing short-term knowledge. It offers no advantage, however, in improving long-term retention of knowledge or in promoting compliance with medical regimens. A strength of video is role-modeling. When applied to well-defined, self-limited stressful situations, role-modeling in video decreases patients' anxiety, pain, and sympathetic arousal while increasing knowledge, cooperation, and coping ability. These effects may carry over for patients to less structured but similarly stressful situations. PMID- 3050103 TI - Blood flow in obstetrics using Doppler ultrasound. AB - We describe the history of obstetric blood flow research in the human fetal circulation (1978-1982) using both continuous wave (CW) Doppler and pulsed (PW) Doppler with linear array scanning. After the initial work (mainly in Ireland, Australia, Sweden, The Netherlands and the UK) it was recognized that CW Doppler velocity waveforms were of diagnostic significance and yielded as much useful clinical information as more complex PW duplex systems. PW duplex continues to be a useful research tool where accurate Doppler sampling or mean velocity measurement is required. The measurement of fetal Doppler velocity waveforms is clinically useful in the evaluation of the 'small-for-dates' fetus and abnormalities of cardiac anatomy or rhythm. Simple CW pencil probe study of the maternal utero-placental vasculature distinguishes normal from abnormal placentation and may be useful as a screening test to predict hypertensive disease of pregnancy. PMID- 3050104 TI - Infections in continuous ambulatory peritoneal dialysis. PMID- 3050105 TI - Bactericidal activity of human serum against strains of Klebsiella from different sources. AB - One hundred and eighty strains of Klebsiella were tested for sensitivity to pooled normal human serum, capsular type and faecal coliform reaction. Strains of K. pneumoniae isolated from clinical infections were more likely to be serum resistant than those from the gut flora or environmental sources. For K. oxytoca strains, there was no significant correlation between serum sensitivity and source of isolation. Of 60 strains of K. oxytoca tested, 19 (32%) were serum resistant, as were 27 (23%) of 120 strains of K. pneumoniae. Strains of K. pneumoniae from human sources gave positive reactions in the faecal coliform test more frequently than strains from environmental sources. There was no correlation between a positive faecal coliform reaction and resistance to the bactericidal effect of human serum. Strains of capsular type K21 were isolated more frequently from clinical infections and were more often serum resistant than other strains. Serum resistance appears to be a virulence factor in strains of K. pneumoniae but does not account for the difference in pathogenicity between K. pneumoniae and K. oxytoca. PMID- 3050106 TI - Penicillin tolerance among oral streptococci. AB - Penicillin tolerance was elicited in 18 of 46 strains of viridans streptococci isolated from the mouths of 19 of 20 healthy subjects and in 31 of 54 consecutive blood-culture isolates of streptococci. Enterococci and Streptococcus sanguis were the organisms most frequently tolerant but the property was also common among isolates of S. mutans, S. mitior and Lancefield Group G streptococci. Pneumococci and S. salivarius were rarely tolerant. When incubated with penicillin at 5 x MIC in batch or continuous cultures, both tolerant and sensitive strains of oral streptococci declined in number less rapidly than S. pyogenes. However, combinations of penicillin and gentamicin killed tolerant and sensitive oral streptococci. PMID- 3050107 TI - Experimental arthritis induced by atypical strains of Streptococcus pyogenes. AB - Experimental arthritis was induced in Sprague-Dawley rats by intraarticular injection of whole-cell sonicates, heat-killed cells and cell-wall preparations of typical and atypical strains of Group-A streptococci (Streptococcus pyogenes). The non-haemolytic nitrosoguanidine-derived mutant and the naturally occurring Lowry strain induced a similar but less severe form of arthritis. Direct immunofluorescent staining demonstrated maximum fluorescence in the sections of articular joint taken 60 days after injection. The level of immune complexes increased for up to 90 days after injection of cell walls or whole-cell sonicates and correlated well with the development of the chronic stage of arthritis observed in haematoxylin and eosin and fluorescence staining of thin tissue sections. PMID- 3050108 TI - Size and homology of the genomes of leprosy-derived corynebacteria, Mycobacterium leprae, and other corynebacteria and mycobacteria. AB - The genomes of Mycobacterium leprae and leprosy-derived corynebacteria (LDC), which have a similar base composition of guanine + cytosine 56 mol %, have been compared with those of reference bacteria of the CMN group (genera Corynebacterium, Mycobacterium, Nocardia). Genome sizes of three LDC strains were (1.2-2.5) x 10(6) base pairs. DNA from four of seven LDC strains examined had homology levels greater than 60%. Two other strains had a homology of 40% when compared with the CMN strains and one strain was distinctly different. The DNA from all seven LDC strains gave 0.3-18% hybridisation with that of M. leprae, 5 16% with reference corynebacteria, 5-12% with M. bovis, and 2-8% with Nocardia caviae. The small size of the LDC genome and its unrelatedness to those of M. leprae and organisms of the CMN group shows the uniqueness of LDC. PMID- 3050110 TI - Arnold Ashley Miles. 20 March 1904-11 February 1988. PMID- 3050109 TI - The role of diarrhoeagenic Escherichia coli in acute diarrhoeal diseases in Bandar-Abbas, Iran. AB - The prevalance of different types of diarrhoea-producing Escherichia coli was measured in 273 patients attending 12 out-patient clinics in Bandar-Abbas, State of Hormozgan, Iran, during March 1984. Enteropathogenic E. coli (EPEC) belonging to 12 different serogroups, of which O128 and O126 were the most common, were found in almost 31% of the patients. Enterotoxigenic strains of E. coli (ETEC) were the next most frequent group (21.9%); among these, 36 (60%) strains produced heat-stable enterotoxin (ST), 14 (23.3%) strains produced both heat-labile enterotoxin (LT) and ST, and 10 (16.7%) strains produced LT only. The same pattern of toxigenicity was observed among the EPEC isolates. Ten of the 12 serogroups encountered in this study contained toxin producers, amongst which strains producing ST were dominant. Enteroinvasive E. coli (EIEC) strains were not isolated. These findings suggest that enterotoxin-producing E. coli may be an important cause of diarrhoea in this part of Iran. PMID- 3050111 TI - The in-vitro effect of a titanium implant on oral microflora: comparison with other metallic compounds. AB - Dental implant research has been mostly concerned with the biocompatibility of materials for implantation. In this study the effects of titanium dioxide and other metallic salts on seven bacterial species commonly found in dental plaque, two which are uncommon, and a yeast, were determined by agar incorporation and diffusion techniques, and compared with the effects of a titanium implant abutment. Neither the titanium dioxide nor the implant abutment demonstrated any inhibitory activity, although other compounds such as cobalt used in dental alloys had some effects. PMID- 3050112 TI - Opsonophagocytosis of Campylobacter pylori. AB - The opsonic activity of human serum from various sources against Campylobacter pylori was compared. All sera, whether from control subjects with no symptoms of gastritis or peptic ulceration, or from symptomatic patients from whom C. pylori had or had not been isolated, opsonised C. pylori equally well. Opsonisation depended on the alternative pathway of complement activation but not on antibody. These findings suggest that antibody plays no role in protection against C. pylori and that the presence of antibody in patients' sera is mainly of diagnostic value. PMID- 3050113 TI - Use of enzymes in typing group A beta-haemolytic streptococci. AB - A series of 698 strains of group A beta-haemolytic streptococci (GAS) isolated from children with streptococcal pyoderma was tested for production of serum opacity factor (OF) and nicotinamide adenine dinucleotide glycohydrolase (NADase). OF was produced by 37% of strains and 40% produced NADase. Classification based on various combinations of OF and NADase reactions showed that 58% belonged to enzyme group el and 34% to e2. Correlation with T and M types showed the possible use of this means of classification as an epidemiological marker for GAS. The specificity of such a system in the further classification of various T types of GAS in epidemiological studies, in the light of antigenic variation among M types, is described. PMID- 3050114 TI - Antimicrobial resistance in Haemophilus influenzae. PMID- 3050115 TI - Effects of chlorpromazine, berberine and verapamil on Escherichia coli heat labile enterotoxin-induced intestinal hypersecretion in rabbit ileal loops. AB - The effects of chlorpromazine (CPZ), berberine and verapamil on intestinal hypersecretion in the rabbit ileal loop model by the heat-labile enterotoxin (LT) of Escherichia coli were studied in relation to their ability to inhibit the stimulation of intestinal adenylate cyclase (AC) by LT. CPZ 5 mg by the intraluminal (i.1.) route and 4 mg/kg by the intramuscular (i.m.) route significantly reduced LT-induced intestinal hypersecretion. Berberine (10 mg) exerted an inhibitory effect, but only after i.1. administration, whereas verapamil did not exert any significant inhibitory effect when administered either i.1. (2.5 mg) or i.m. (4 mg/kg). At concentrations of (0.17-1.34) X 10(-3) M CPZ, the anti-secretory effect of CPZ correlated with its inhibitory effect on rabbit LT-stimulated intestinal AC. Inhibition of cAMP synthesis was probably not involved in the mechanism of action of the two other substances. These results indicate that CPZ and phenothiazines in general are efficient drugs for reducing LT-induced intestinal hypersecretion and could represent a model for synthesis of new anti-secretory drugs with no tranquiliser side effects. PMID- 3050116 TI - Effects of pyridine nucleotides on the gating of ATP-sensitive potassium channels in insulin-secreting cells. AB - The single-channel current recording technique has been used to study the influences that the pyridine nucleotides NAD, NADH, NADP and NADPH have on the gating of ATP-sensitive K+ channels in an insulin-secreting cell line (RINm5F). The effects of the nucleotides were studied at the intracellular surface using either excised inside-out membrane patches or permeabilized cells. All four pyridine nucleotides were found to evoke similar effects. At low concentrations, 100 microM and less, each promoted channel opening whereas high concentrations, 500 microM and above, evoked channel closure. The degree of K+ channel activation by pyridine nucleotides (low conc.) was found to be similar to that evoked by the same concentrations of ADP or GTP, whereas the degree of K+ channel inhibition (high conc.) was less marked than that evoked by the same concentrations of ATP, and never resulted in refreshment of K+ channels following removal. The effects of NAD, NADH, NADP and NADPH seemed to interact with those of ATP and ADP. In the presence of 1 mM ADP and 4 mM ATP, 10 to 100 microM concentrations of the pyridine nucleotides could not evoke channel opening, whereas concentrations of 500 microM and above were found to evoke channel closure. In the presence of 2 mM ATP and 0.5 mM ADP, however, 10 to 100 microM concentrations of the pyridine nucleotides were able to activate K+ channels. PMID- 3050118 TI - Desmoid tumors: report of a case responsive to antiestrogen and review of the literature. PMID- 3050117 TI - Type A behavior as a general risk factor for physical disorder. AB - A behavior pattern characterized by excessive competitiveness, impatience, hostility, and time urgency, known as Type A, has typically been investigated as a risk factor for coronary heart disease. The present paper evaluates the Type A pattern as a general risk factor for a wide variety of physical disorders. Research on Type A as a moderator of the effects of life stress on health is also reviewed. When Type A or physical health is measured with objective indicators, Type A does not emerge as a general risk factor for illness, with the following exceptions: Type A's are more likely to have accidents, to die from accidents or violence, and to incur cerebrovascular and peripheral atherosclerosis. In contrast, research relying on self-report measures of Type A and symptomatology find a consistent link between Type A behavior and a variety of minor illness and symptoms. There is little support for the notion that Type A is a potentiator of the effects of life events stress. PMID- 3050119 TI - Probing co-operative DNA-binding in vivo. The lac O1:O3 interaction. AB - The lac primary (O1) and weak upstream pseudo (O3) operators contained on a plasmid were footprinted in vivo in order to determine whether they act co operatively in binding lac repressor in the cell. The occupancy at O3 by lac repressor was substantially reduced upon deletion of the lac primary operator, demonstrating co-operativity at a distance. Plots of operator occupancy versus active repressor concentration were obtained for each operator by treating the cells with different amounts of the lac inducer isopropyl-beta-D-thiogalactoside and probing lac repressor binding. This analysis can be used to obtain relative binding constants in vivo and demonstrates that O3 binds repressor only 10.3-fold less tightly than O1 in their co-operative interaction. The removal of DNA torsional tension in vivo by the use of coumermycin leads to the same loss of binding at O3 as does deleting O1. These in-vivo results are analogous to the in vitro situation, where O3 binds repressor strongly in a DNA repression loop only on supercoiled templates. PMID- 3050120 TI - Estimation of in-vivo miscoding rates. AB - The replication of premutagenic DNA lesions generates mutant progeny in patterns that distinguish lesions that rarely produce a mutation per DNA replication from those that frequently do so. The quantitative aspects of this distinction were tested in studies of heat-mutagenized bacteriophage T4. Previous T4 studies had demonstrated that transition mutations produced at G.C base-pairs depended upon heat-induced DNA lesions distinct from those responsible for transversions at G.C pairs. In this study the transversion mutations are shown to arise in patterns predicted for mutations produced from lesions that miscode rarely (fewer than 10% per replication). In contrast, the transition mutations arise in patterns predicted for mutations produced from lesions that miscode at about 20 to 60% per replication. The fact that the two classes of DNA lesions are distinguishable as predicted by the quantitative model suggests that such studies may in general be useful in quantifying the behavior of mutation-generating DNA lesions. The method employed also estimates the frequency of premutagenic lesions in DNA. PMID- 3050121 TI - Mapping and sequencing of mutations in the Escherichia coli rpoB gene that lead to rifampicin resistance. AB - Rifampicin is an antibiotic that inhibits the function of RNA polymerase in eubacteria. Mutations affecting the beta subunit of RNA polymerase can confer resistance to rifampicin. A large number of rifampicin-resistant (hereafter called Rifr) mutants have been isolated in Escherichia coli to probe the involvement of RNA polymerase in a variety of physiological processes. We have undertaken a comprehensive analysis of Rifr mutations to identify their structural and functional effects on RNA polymerase. Forty-two Rifr isolates with a variety of phenotypes were mapped to defined intervals within the rpoB gene using a set of deletions of the rpoB gene. The mutations were sequenced. Seventeen mutational alterations affecting 14 amino acid residues were identified. These alleles are located in three distinct clusters in the center of the rpoB gene. We discuss the implications of our results with regards to the structure of the rifampicin binding site. PMID- 3050123 TI - Characterization of the termination phenotypes of rifampicin-resistant mutants. AB - Rifampicin-resistant (Rifr) mutations map in the rpoB gene encoding the beta subunit of Escherichia coli RNA polymerase. We have examined the effect of each of the 17 sequenced Rifr mutations in our collection on transcription termination. The effect of each Rifr mutation was measured at three types of terminators: simple terminators requiring only RNA polymerase to terminate in vitro, and complex terminators requiring either Rho or Tau for in-vitro termination. Almost every Rifr allele examined (14/17) affected readthrough at one or more of these terminators. We found that mutations with similar termination phenotypes were clustered suggesting functional specialization within the region of rpoB defined by the Rifr mutations. The interaction of the Rifr mutations with the defective rho15 allele was also investigated. Only two Rifr mutations suppress the termination defect of rho15 strains. We discuss models to explain how this region of the beta polypeptide might be involved in the process of transcription termination. PMID- 3050122 TI - Investigation of the mechanism of active site coupling in the pyruvate dehydrogenase multienzyme complex of Escherichia coli by protein engineering. AB - Site-directed mutagenesis of the aceF gene of Escherichia coli was used to generate a nested set of deletions in the long (alanine + proline)-rich sequence that separates the lipoyl domain from the dihydrolipoamide dehydrogenase-binding domain in the "one-lipoyl domain" dihydrolipoamide acetyltransferase polypeptide chains of a pyruvate dehydrogenase multienzyme complex. The deletions reduced the number of residues in this sequence successively from 32 to 20, 13, 7 and just 1 residue. In all instances, pyruvate dehydrogenase complexes were still assembled in vivo around cores containing the deleted chains, and those with the two shortest deletions were essentially fully active. However, the two most severe deletions caused falls of 50% or more in specific catalytic activity. Similarly, although shortening the interdomain sequence to 20 residues left the system of active-site coupling unimpaired, cutting it to 13 residues or less caused substantial falls in the reductive acetylation of the lipoyl domains and corresponding losses of active-site coupling. The changes in specific catalytic activity and active-site coupling that accompanied the shortening of the (alanine + proline)-rich segment were reflected in the poorer growth rates of the relevant strains of E. coli on stringent substrates. All these results are consistent with this (alanine + proline)-rich sequence acting as a linker region that facilitates the movements of the lipoyl domains required for full catalytic activity and active-site coupling in the complex. The other two such sequences that separate the additional lipoyl domains in the N-terminal half of the wild-type "three lipoyl domain" dihydrolipoamide acetyltransferase chain are presumed to function similarly. This role is consistent with the conformational flexibility assigned to these segments from previous studies based on 1H nuclear magnetic resonance spectroscopy and protein engineering. PMID- 3050124 TI - Tet repressor-tet operator contacts probed by operator DNA-modification interference studies. AB - Contacts between tet operator DNA and Tet repressor protein are characterized by modification interference studies. The modified DNA fragments are separated into fractions with high, intermediate and low affinities for Tet repressor by polyacrylamide gel electrophoresis. Ethylation of the phosphates with N ethylnitrosourea reveals 12 contacts of a repressor dimer to tet operator. Eight of these contacts appear to be important for Tet repressor binding, as judged by the strong interference at these positions, while four contacts are probably less important. All of the phosphate contacts are located on the same side of the B DNA structure. The sequences of tet operators proposed to interact with the recognition alpha-helix of Tet repressor are TCTATC in three cases and CCTATC in one case. After methylation of N-7 with dimethylsulfate, strong interference is observed at the guanine residues at positions +/- 2. None of the N-7 functions of other guanine residues seems to be involved in tight contacts to Tet repressor. Tet repressor subunits form identical phosphate and guanine N-7 contacts with each half side of the two tet operators indicating twofold dyad symmetry of the complexes. Attempts to analyze the methylation interference at the adenine N-3 sites reveal different results for the operators. Modification of DNA fragments with diethylpyrocarbonate yields hypersensitive sites in the tet operators, indicating different local DNA structures. Carbethoxylation interference studies confirm the contacts at the purines found by methylation interference. All of the sequence-specific protein-DNA contacts detected in this study are centered at the inside four base-pairs in each tet operator half side. The contacts are discussed with respect to the structure of the repressor-operator complex. PMID- 3050125 TI - Stereospecific positioning of the cis-acting sequence with respect to the canonical promoter is required for activation of the ompC gene by a positive regulator, OmpR, in Escherichia coli. AB - Expression of the ompC gene coding for a major outer-membrane protein of Escherichia coli is regulated by a transcriptional activation mechanism that requires the ompR gene product, OmpR. It was demonstrated that multiple OmpR molecules bind to a cis-acting sequence located upstream from the canonical -35 and -10 regions of the ompC promoter. Using an ompC-lacZ fusion gene, the distance between the cis-acting upstream sequence (OmpR-binding site) and the -35 and -10 regions (RNA polymerase-binding site) has been changed. We demonstrated that the ompC transcription was activated in an OmpR-dependent manner even when the cis-acting upstream sequence was separated from the -35 and -10 regions by several turns of the DNA helix, providing that the distance between them was a near-integral multiple of one turn of the DNA helix. Evidence is presented that stereospecific positioning of the cis-acting upstream sequence with respect to the canonical promoter is required for activation of the ompC gene by the positive regulator, OmpR. PMID- 3050126 TI - Promoter recognition by Escherichia coli RNA polymerase. Influence of DNA structure in the spacer separating the -10 and -35 regions. AB - Escherichia coli RNA polymerase contacts promoter DNA at two regions (the -10 and -35 regions) which are separated by a segment of spacer DNA. Previously we showed that base substitutions in the spacer DNA can affect promoter strength both in vitro and in vivo; these results were interpreted to reflect altered structural properties of the substituted DNAs. Here we provide experimental support for this interpretation. The pattern of cleavage of the promoters with Neurospora crassa endonuclease and the reactivity of their guanine residues with dimethyl sulfate (DMS) suggest that the structures of the spacer DNAs in the promoters with altered transcriptional activities are distinct. In addition, the binding of RNA polymerase to the latter promoters induces characteristic enhancements in the extent to which specific guanine residues in the spacer DNAs react with DMS. We propose that for these promoters the substitutions in the spacer DNAs have affected the relative orientation of the -10 and -35 regions. The observed differences in promoter activity then would reflect the requirement for realignment of these regions during the process of open complex formation; we postulate that two such realignments occur. PMID- 3050127 TI - In-vivo studies on the cis-acting replication initiator protein of IncFII plasmid NR1. AB - Using segment-directed mutagenesis, a temperature-sensitive mutant of the gene that encodes the cis-acting RepA1 initiation protein of the IncFII plasmid NR1 was isolated. The mutant protein was unable to promote initiation of plasmid replication in vivo at 42 degrees C. Both the wild-type and the mutant repA1 genes were cloned separately into the high-expression vector plasmid pAS1. In these pAS1-repA1 derivatives, the transcription of the repA1 gene was under the control of the lambda PL promoter, which was regulated by the temperature sensitive lambda cI857 repressor protein. The translation initiation of the repA1 mRNA from these derivatives was mediated by the lambda cII Shine-Dalgarno sequence and initiation codon. The yield of 33,000 Mr RepA1 protein detected on SDS/polyacrylamide gels from Escherichia coli cells containing the pAS1-repA1 derivatives was dependent upon whether the newly synthesized RepA1 was capable of interacting in cis with the downstream NR1 replication origin on the cloned DNA fragment. Mutations in the repA1 gene or deletions of the cis origin region dramatically increased the detectable yield of RepA1 protein. Deletion of the NR1 origin region from the pAS1 derivative containing the wild-type repA1 gene enabled the cis-acting RepA1 protein to complement partially the temperature sensitive repA1 mutant in trans, to increase the copy number in trans of plasmids that contained the NR1 replicon, and to help NR1 derivatives overcome plasmid incompatibility. The trans effects of RepA1 provided by the pAS1-repA1 derivatives that retained the origin in cis were much less significant. RepA1 provided in trans also stimulated the replication of plasmids carrying cloned copies of the NR1 replication origin region regardless of whether the origin was transcribed from an upstream promoter. PMID- 3050128 TI - Effects of mot gene expression on the structure of the flagellar motor. AB - Direct freezing procedures have enabled us to visualize distinctive intramembrane particle ring structures in the cytoplasmic membranes of peritrichously flagellated bacteria by means of freeze-fracture electron microscopy. These structures were identified as flagellar motor components because their distribution matched that of flagella, and because they were absent in non flagellated mutants of Escherichia coli. Particle rings were present in both the Gram-positive Streptococcus and the Gram-negative E. coli. In E. coli, a non functional mocha operon produced flagellated but immotile cells lacking the particle rings. Simultaneous introduction of the motA and motB genes, led to recovery of both motility and the ring structures but neither gene alone was sufficient. The concomitant loss of the rings and motility is consistent with the ring particles having a central role in the flagellar motor. PMID- 3050129 TI - Structure of helical RecA-DNA complexes. III. The structural polarity of RecA filaments and functional polarity in the RecA-mediated strand exchange reaction. AB - The RecA protein of Escherichia coli has been used in vitro to mediate a strand exchange reaction between homologous DNA molecules. A three-dimensional reconstruction of a RecA filament on double-stranded DNA has been previously determined from electron micrographs, and the reconstruction displays a clear axial polarity. The RecA-mediated strand-exchange reaction between a double stranded DNA and a homologous single-stranded DNA that is complexed with a RecA helical polymer proceeds with a known polarity. Using image analysis of electron micrographs, we have determined the relation between the structural polarity of RecA filaments and the 3' and 5' polarity of single-stranded DNA. Thus, the structural polarity of RecA filaments can now be related to the direction in which the RecA-mediated strand-exchange reaction advances along the complexed single-stranded DNA. PMID- 3050130 TI - Testing for intron function in the essential Saccharomyces cerevisiae tRNA(SerUCG) gene. AB - The gene sup61+, which codes for the essential Saccharomyces cerevisiae tRNA(SerUCG), is the only single-copy tRNA gene in this organism know to contain an intron. To assess the role of this intron in tRNA gene expression, an intron deleted sup61+ gene was constructed in vitro and introduced into the yeast genome. Isogenic intron- and intron+ strains were found to be indistinguishable by criteria that include growth rates, ability to undergo meiosis, levels of mature tRNA(SerUCG) transcribed in vivo, and the suppressor efficiency of amber- and ochre-specific alleles of this gene. PMID- 3050131 TI - trp repressor interactions with the trp aroH and trpR operators. Comparison of repressor binding in vitro and repression in vivo. AB - Interaction of the Escherichia coli trp repressor with the promoter-operator regions of the trp, aroH and trpR operons was studied in vivo and in vitro. The three operators have similar, but non-identical, sequences; each operator is located in a different segment of its respective promoter. In vivo repression of the three operons was measured using single-copy gene fusions to lacZ. The extent of repression varied from 300-fold for the trp operon, to sixfold for the aroH operon and threefold for the trpR operon. To determine whether differential binding of repressor to the three operators was responsible for the differences in repression observed in vivo, three in vitro binding assays were employed. Restriction-site protection, gel retardation and DNase footprinting analyses revealed that repressor binds to the three operators with almost equal affinity. It was also shown in an in vivo competition assay that repressor binds approximately equally well to each of the three operators. It is proposed that the differential regulation observed in vivo may be due to the different relative locations of the three operators within their respective promoters. PMID- 3050132 TI - Allele-specific malE mutations that restore interactions between maltose-binding protein and the inner-membrane components of the maltose transport system. AB - Active accumulation of maltose and maltodextrins by Escherichia coli depends on an outer-membrane protein. LamB, a periplasmic maltose-binding protein (MalE, MBP) and three inner-membrane proteins, MalF, MalG and MalK. MalF and MalG are integral transmembrane proteins, while MalK is associated with the inner aspect of the cytoplasmic membrane via an interaction with MalG. Previously we have shown that MBP is essential for movement of maltose across the inner membrane. We have taken advantage of malF and malG mutants in which MBP interacts improperly with the membrane proteins. We describe the properties of malE mutations in which a proper interaction between MBP and defective MalF and MalG proteins has been restored. We found that these malE suppressor mutations are able to restore transport activity in an allele-specific manner. That is, a given malE mutation restores transport activity to different extents in different malF and malG mutants. Since both malF and malG mutations could be suppressed by allele specific malE suppressors, we propose that, in wild-type bacteria, MBP interacts with sites on both MalF and MalG during active transport. The locations of different malE suppressor mutations indicate specific regions on MBP that are important for interacting with MalF and MalG. PMID- 3050133 TI - Structure of light-harvesting chlorophyll a/b protein complex from plant photosynthetic membranes at 7 A resolution in projection. AB - The structure of thin three-dimensional crystals of the light-harvesting chlorophyll a/b protein complex, an integral membrane protein from the photosynthetic membrane of chloroplasts, has been determined at 7 A (1 A = 0.1 nm) resolution in projection. The structure analysis was carried out by image processing of low-dose electron micrographs, and electron diffraction of thin three-dimensional crystals preserved in tannin. The three-dimensional crystals appeared to be stacks of two-dimensional crystals having p321 symmetry. Results of the image analysis indicated that the crystals were disordered, due to random translational displacement of stacked layers. This was established by a translation search routine that used the low-resolution projection of a single layer as a reference. The reference map was derived from the symmetrized average of two images that showed features consistent with the projected structure of negatively stained two-dimensional crystals. The phase shift resulting from the displacement of each layer was corrected. Phase shifts were then refined by minimizing the phase residual, bringing all layers to the same phase origin. Refined phases from different images were in agreement and reliable to 7 A resolution. A projection map was generated from the averaged phases and electron diffraction amplitudes. The map showed that the complex was a trimer composed of three protein monomers related by 3-fold symmetry. The projected density within the protein monomer suggested membrane-spanning alpha-helices roughly perpendicular to the crystal plane. The density in the centre and on the periphery of the trimeric complex was lower than that of the protein, indicating that this region contained low-density matter, such as lipids and antenna chlorophylls. PMID- 3050135 TI - Molecular biology in cardiology, a paradigmatic shift. AB - Control of the performance of the heart can now be understood in terms of three paradigms: beat-to-beat, length-dependent regulation (Starling's Law of the Heart); short term regulation by biochemical changes within the cardiac cell (excitation-contraction coupling, myocardial contractility); and long term regulation by altered gene expression (molecular biology). The latter may also compensate for inhomogeneities in ventricular function. According to Kuhn's theory, the more recent of these paradigms should provide a better and more complete picture of cardiac function. However, because each of these paradigms explains distinct aspects of cardiac regulation that are not mutually exclusive, this evolution fails to support Kuhn's view that science progresses as a series of revolutionary advances. Instead, it is only by understanding all of these paradigms that we can begin to grasp their complex interactions in governing cardiac function, and so gain insight into these interwoven control mechanisms. It seems likely that this is not an uncommon pattern, and that future work will add new paradigms to the complex body of knowledge of cardiac regulation without invalidating any of the paradigms described in this editorial. PMID- 3050136 TI - Laudatio for John Money on the occasion of the 1988 Masters and Johnson award of the Society for Sex Therapy and Research. PMID- 3050137 TI - The ethics of pornography in the era of AIDS. AB - Since the inception of sexology as an academic discipline a century ago, the boundary between sexology, the science, and sexosophy, the philosophy of sex, has been poorly demarcated, especially with respect to the principles of sex-reform movements. Several early 20th centruy sexologists overtly espoused the principles of eugenics reform, which, in the 1930s, Hitler used against them. A large proportion of today's sex therapists, researchers, and educators are among "those who cannot remember the past" and are, therefore, according to Santayana, "condemned to repeat it." That is to say, they follow the example of eugenics reformers by adhering to explanatory principles as if they were apolictically indisputable; whereas, they are, in fact, dangerously political professional platitudes for the criminalization of sex. One such platitude is that pornography is dehumanizing and a socially contagious criminal offense. Social-contagion theory had its origin in Tissot's 18th century revival of semen-conservation theory. In America, Tissot's antisexual health-reform ideas were transmitted by Graham to Kellogg and Comstock. The Comstock Laws of 1873 are still extant and are the basis of contemporary judicial, academic, and public misconceptions regarding the social contagiousness of pornography. These misconceptions render the nation incapable of using pornography constructively in a program of sex safety to prevent AIDS infection, especially among newly pubertal adolescents and young adults. The model to follow has already been provided by gay JACK and JO masturbation clubs. PMID- 3050134 TI - Barnase and barstar. Expression of its cloned inhibitor permits expression of a cloned ribonuclease. AB - Barnase is the extracellular ribonuclease of Bacillus amyloliquefaciens and barstar its specific intracellular inhibitor. The gene for barstar has now been cloned and sequenced. When the wild-type gene for barnase is reconstructed from its previously cloned parts on the same plasmid as the barstar gene, the lethal effect of its expression is suppressed. A plasmid has been devised which directs the secretion of 100 mg per active barnase liter by Escherichia coli and another which provides large (500 to 1000 mg/l) yields of barstar. The structure of these plasmids and the derived 89 amino acid sequence of barstar are reported. PMID- 3050138 TI - Perspectives of comparing risks of environmental carcinogens. AB - In 1987, investigators (Ames et al.) concluded that the risks of man-made industrial carcinogens and pesticides (outside of the workplace) are trivial compared with the risks of naturally occurring carcinogens found mostly in the diet. They used a ranking system based on human exposure and rodent potency (HERP) data to arrive at this conclusion. As a result, they recommend that regulatory agencies, such as the Environmental Protection Agency and the Food and Drug Administration, base their priorities in this area on their HERP system. We analyzed the assumptions and data set upon which the HERPs were based, concluding that such a simplified approach to set public health policy is inappropriate given the underlying uncertainties. However, we note that when comparisons are consistently based on estimates of average daily exposure to common carcinogens, the HERP scores of many man-made pollutants are comparable to those of naturally occurring carcinogens in the diet. PMID- 3050139 TI - Successful clinical use of an anti-HLA-DR monoclonal antibody for autologous bone marrow transplantation. AB - It has been widely assumed that anti-HLA-DR antibodies react with pluripotent stem cells and cannot be used in bone marrow purging. We report a case of non Hodgkin's lymphoma in which an anti-HLA-DR antibody (AB4) was used for immunomagnetic purging and the subsequent autologous bone marrow transplantation resulted in rapid marrow engraftment with no serious complications. The results indicate that the AB4 antibody, which binds to an antigen encoded by the B3 gene of the DR region, can be safely used in the clinic in the purging of bone marrow from patients with AB4-positive tumors (non-T-cell acute lymphocytic leukemia, non-Hodgkin's lymphoma, and some cases of acute myelogenous leukemia. PMID- 3050140 TI - Testicular cancer: the quest continues. AB - Treatment for testicular cancer has dramatically improved during the last 15 years. Much of this success has come about because of improved staging and operative techniques but, most importantly, through the introduction of successful systemic chemotherapy. Nonetheless, relevant issues still remain to be addressed in regard to the optimal therapy for patients with germ cell neoplasms. Included in these issues is delineating the most effective but minimally morbid treatment for patients with early-stage and low-volume metastatic disease while continuing to create innovative treatment approaches for poor-risk patients with metastatic disease. The unique association of primary mediastinal germ cell neoplasms with the development of non-germ cell cancers and Klinefelter's syndrome may provide some early clues for the determination of factors controlling differentiation. These observations issue a challenge to both clinical and preclinical researchers involved in the study of this neoplasm. PMID- 3050142 TI - Development of hepatocytes in the pancreas of hamsters treated with 2,3,7,8 tetrachlorodibenzo-p-dioxin. AB - Transdifferentiation is a process in which one differentiated cell type is converted to another. A unique example of transdifferentiation is the development of hepatocytes from pancreatic cells in adult hamsters and rats. In this communication we report the induction of pancreatic hepatocytes in hamsters that were given 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Two or 6 intraperitoneal (ip) injections of TCDD at a dose of 100 micrograms/kg body weight at 4-wk intervals induced pancreatic hepatocytes in 75% and 89% of the animals respectively. In animals given only two doses of TCDD each pancreas contained one to two hepatic foci, whereas when six injections were administered multiple hepatic foci were observed. By hematoxylin and eosin stain and by periodic acid Schiff stain, the pancreatic hepatocytes were morphologically identical to those in normal liver. Although the exact mechanism by which TCDD induces the transformation is not clear, it is conceivable that TCDD acting through receptor mediated mechanisms is activating the repressed liver-specific genes in the pancreas. PMID- 3050141 TI - Urinary cadmium and beta 2-microglobulin: correlation with nutrition and smoking history. AB - Urinary cadmium and beta 2-microglobulin concentrations from approximately 1000 samples from the general adult U.S. population, collected as part of the National Health and Nutritional Examination Survey II (NHANES II), were related to nutritional and smoking history of the individuals. Urinary cadmium concentration was negatively correlated with dietary iron (significance level of 0.0065), negatively correlated with dietary calcium (significance level of less than 0.0001), and significantly (level of less than 0.001) higher in past or present smokers than in those who had never smoked. The results suggest increased cadmium absorption in the presence of low dietary intake of iron, low dietary intake of calcium, and cigarette smoking in the general population of the United States. PMID- 3050143 TI - Changes in T-lymphocyte subpopulations and natural killer cells following exposure to ambient levels of nitrogen dioxide. AB - The effects of ambient levels of NO2 on murine splenic T-lymphocyte subpopulations and natural killer cells were investigated. AKR/cum and C57BL/6J mice were exposed, in inhalation chambers, to 0.25 ppm NO2 for 7 wk and 0.35 ppm NO2 for 12 wk, respectively. Monoclonal antibody technology was used in conjunction with fluorescence-activated cell sorter (FACS) analysis to detect quantitative changes in total T-lymphocytes (Thy-1.2-positive), mature T lymphocytes (Lyt-1-positive), T-helper/inducer lymphocytes (L3T4-positive), T cytotoxic/suppressor lymphocytes (Lyt-2-positive), and natural killer cells (asialo GM1-positive). Percentages of all T-lymphocyte subpopulations tested and natural killer cells were lower in spleens of mice exposed to NO2 compared to filtered-air controls. This is the first report providing evidence linking alterations in T-lymphocyte subpopulations and natural killer cells to NO2 exposure at ambient levels. Changes in T-lymphocyte subpopulations detected by FACS and correlated to impaired immune function may provide an extremely sensitive means of demonstrating NO2-induced changes in the immune system. PMID- 3050145 TI - Changes in extravascular lung water and fatty acids in a hyperdynamic canine model of sepsis. AB - Several mediators, including fatty acids, have been postulated to induce the increase in permeability of the pulmonary endothelium and subsequent accumulation of extravascular lung water (EVLW) which are generally considered to be among the first events in the development of adult respiratory distress syndrome. In a canine model of hyperdynamic sepsis (hemodynamically similar to human sepsis) changes in EVLW and in concentrations of different fatty acids in the aortic and pulmonary arterial blood were measured. Two days after induction of sepsis, EVLW increased from 6.6 +/- 0.6 to 9.2 +/- 1.0 ml/kg, and the pulmonary arterial concentration of oleic acid increased from 52 +/- 4 to 73 +/- 5 mg/dl. Three days after induction of sepsis, EVLW increased further to 14.4 +/- 3.8 ml/kg and the mean concentration of oleic acid increased to 74 +/- 7 mg/dl. Twenty-four hours later, both EVLW and the mean pulmonary arterial concentration of oleic acid were not different from basal. We postulate that oleic acid, a known inducer of experimental ARDS, is one of the mediators of endothelial damage of the lung during sepsis. PMID- 3050144 TI - Mutagenicity test of cefodizime sodium. AB - The mutagenicity of cefodizime sodium (THR-221) was investigated by the reverse mutation test using seven bacterial strains (Salmonella typhimurium strains TA100, TA98, TA1535, TA1537 and TA1538, and Escherichia coli strains WP2 and WP2uvrA) and the chromosomal aberration test with cultured Chinese hamster lung (CHL) cells. The reverse mutation test was carried out in dose range from 0.0025 to 5.0 micrograms/plate in the absence and presence of mammalian metabolic activation system. Antibacterial effects were observed at concentrations more than 0.25 microgram/plate, and no significant increases in the number of revertants were observed at the dose levels where antibacterial effects were not detected. THR-221 caused no increases in the number of chromosomal aberrants at dose levels of 0.75, 1.5, 3.0 and 6.0 mg/ml in the absence and presence of the metabolic activation. These results indicate that THR-221 has no mutagenic activity. PMID- 3050146 TI - Epidemiology of toxoplasmosis in pregnant women in Tabasco, Mexico. PMID- 3050148 TI - A single-base change in gene 10 of bacteriophage T7 permits growth on Shigella sonnei. AB - Bacteriophage T7 can extend its host range to include Shigella sonnei D2 371-48 by a mutation called ss found in the T7 major capsid protein, the gene 10 product. We show that a single A-to-C transversion at position 23150 in the T7 genome is responsible for the T7 ss mutant phenotype that allows the phage to avoid DNA degradation and undergo productive infection. The ss mutation causes an amino acid substitution of proline for glutamine at position 61 of the 344-amino acid T7 major capsid protein. PMID- 3050147 TI - Expression of bicistronic measles virus P/C mRNA by using hybrid adenoviruses: levels of C protein synthesized in vivo are unaffected by the presence or absence of the upstream P initiator codon. AB - The measles virus (MV) P/C mRNA is functionally bicistronic. Translation is presumed to initiate at both the first and second 5'-proximal AUG codons, leading, respectively, to synthesis of the P and C polypeptides from different overlapping reading frames. To study the function and differential expression of these polypeptides, we have constructed hybrid human adenoviruses capable of expressing high levels of P and C together or of C alone. Cloned cDNA corresponding to the MV P/C gene was coupled to the adenovirus type 2 (Ad2) major late promoter, most of the Ad2 tripartite leader sequence, and the simian virus 40 3'-end processing signal and then used to replace most of the E1a-E1b region of the Ad5 genome in two hybrid adenoviruses: one (Ad5MV/PC13) which contained both 5'-proximal AUG codons of the P/C mRNA and another (Ad5MV/C3) which retained only the second. The sequence context for the P protein initiator AUG codon in Ad5MV/PC13 was made more favorable (GAGAUGG) than the relatively unfavorable context (CCGAUGG) seen in the native MV P/C mRNA. After infection of 293 cells (which provide complementary E1a-E1b functions), both viruses directed equal amounts of P/C-specific mRNA transcription. Ad5MV/PC13 directed the synthesis of both P and C proteins, while Ad5MV/C3 directed the synthesis of C protein alone. Ad5-expressed P protein was phosphorylated, while C was not. C protein had a similar diffuse cytoplasmic localization in both MV and Ad5-infected cells. Ad5MV/C3 and Ad5MV/PC13 directed equal amounts of C protein expression in 293 cells at a level approximately 15 times greater than that seen in MV-infected cells. Thus the level of C protein expression was unaffected by the presence or absence of an out-of-frame upstream AUG codon in a favorable sequence context. This observation cannot be explained by the scanning model for ribosomal initiation and suggests that ribosomes may be binding directly at an internal mRNA site at or near the initiator AUG codon for the C protein. PMID- 3050150 TI - Evidence for cyclophosphamide-induced transitional cell carcinoma in a renal transplant patient. AB - Cyclophosphamide treatment has been associated with bladder cancer in a number of case reports but no causal relationship has been proved since nearly all of these patients were treated with the drug for malignant disease. We describe a patient who received cyclophosphamide after cadaveric renal transplantation to prevent rejection. Transitional cell carcinoma developed in the native bladder and in the donor transplanted ureter (20-year-old donor) 13 years later despite no identifiable risk factors. This case strengthens the argument that cyclophosphamide has a carcinogenic potential on the urinary tract epithelium. PMID- 3050151 TI - Ureteral involvement in the Churg-Strauss syndrome: a case report. AB - We report a case of the Churg-Strauss syndrome with bilateral ureteral stenosis, secondary obstructive uropathy and anuria. The literature regarding urological implications of this disease is reviewed, and the surgical and medical management of our case is detailed. PMID- 3050149 TI - Processing of in vitro-synthesized gag precursor proteins of human immunodeficiency virus (HIV) type 1 by HIV proteinase generated in Escherichia coli. AB - We expressed the gag and proteinase regions of human immunodeficiency virus (HIV) type 1 by transcription and translation in vitro. A synthetic RNA spanning the gag and pro domains gave primarily the unprocessed capsid precursor pr53. Efficient cleavage of this precursor was observed when the gag and pro domains were placed in the same translational reading frame, yielding equimolar amounts of the gag protein and of proteinase (PR). Expression of HIV type 1 PR in Escherichia coli as a fusion protein gave rapid autocatalytic processing to an HIV-specific protein of approximately 11 kilodaltons. HIV PR generated in E. coli specifically induced cleavage of the HIV capsid precursor, whereas deletion of the carboxy-terminal 17 amino acids of the proteinase rendered it inactive. Inhibitor studies showed that the enzyme was insensitive to inhibitors of serine and cysteine proteinases and metalloproteinases and was inhibited only by a very high concentration (1 mM) of pepstatin A. PMID- 3050153 TI - Aztreonam: critical evaluation of the first monobactam antibiotic in treatment of urinary tract infections. PMID- 3050152 TI - Reduction of infection stones in rats by combined antibiotic and phosphocitrate therapy. AB - The potential of phosphocitrate to inhibit infection stones in rats when combined with an antibiotic was studied. A significant reduction occurred in both the number and weight of recovered stones from rats receiving combined treatment with amoxycillin (50 mg./kg. body wt./day and phosphocitrate (112 mumol./kg. body wt./day) for four weeks. The inhibitory responses were attributed to the intact phosphocitrate molecule as administration of citrate in equimolar concentrations did not mimic the observed effects of phosphocitrate. In comparison with non infected controls, antibiotic treatment alone failed to eliminate total stone growth. However, composition of the stone reverted from predominantly struvite to a mixture of struvite and newberyite as urinary parameters normalized. The studies highlight the usefulness of phosphocitrate to restrict magnesium salt deposition in vivo. PMID- 3050154 TI - Unilateral renal agenesis with or without ipsilateral adrenal agenesis. AB - We analyzed 7 patients with unilateral renal agenesis. The coexistence of unilateral renal agenesis with ipsilateral adrenal agenesis was confirmed surgically in 2 patients, 1 of whom had clinical features of Cushing's syndrome owing to adrenocortical adenoma of a solitary adrenal gland. It is imperative to suspect abnormalities of the ipsilateral adrenal gland when unilateral renal agenesis is encountered. Unilateral renal agenesis usually is asymptomatic. Recognition of this anomaly is essential for early correct diagnosis. PMID- 3050155 TI - Blood group isoantigen deletion and chromosomal abnormalities in bladder cancer. AB - The presence or absence of blood group isoantigens in 78 patients with transitional cell carcinoma of the bladder was correlated with tumor stage and grade, and results of chromosomal analysis. For blood group isoantigen detection the indirect immunoperoxidase method with monoclonal antibodies to A, B and H antigen was used. In 51 per cent of the 59 superficial tumors blood group isoantigens were demonstrable, whereas all deeper infiltrating and higher grade tumors were negative. However in superficial tumors the mode of blood group isoantigen expression did not correlate significantly with tumor recurrence and progression. A consistent correlation was demonstrated among chromosomal numbers, tumor grade and clinical course. The chromosomal abnormalities found and mode of blood group isoantigen expression, even in combination, had no prognostic value additional to the grading criteria used. PMID- 3050156 TI - Treatment of hemospermia caused by dilated seminal vesicles by direct drug injection guided by ultrasonography. AB - Bilateral seminal vesicle puncture and injection of drugs with ultrasound guidance were performed in patients with hemospermia resistant to conservative therapy and with dilated seminal vesicles. Of 7 patients 6 had resolution of hemospermia for 2 to 3 months and then relapse. No side effect was noted. PMID- 3050157 TI - Aortic disease associated with pregnancy. AB - Our experience with the management of two patients with life-threatening aortic disease during pregnancy is presented with a review of the literature. One of our patients had intimal disruption caused by trauma; the other had probable Ehlers Danlos type IV syndrome, causing an acute dissection of the descending thoracic aorta and eventually requiring replacement of the aorta from the left subclavian artery to common iliac arteries. The challenge of treating both the pregnant woman and the fetus was managed successfully by an emergent cesarean section followed by Dacron graft replacement of the descending thoracic aorta. The literature reviewed disclosed that aneurysm expansion producing symptoms and dissection is most common during the third trimester and during labor and delivery in patients with or without Marfan's syndrome. Half of the aortic dissections in women less than 40 years of age occur in association with pregnancy. The available evidence indicates that patients with known valvular or aortic disease should have surgical repairs during the first or second trimester and thereafter have delivery by cesarean section. However, patients with acute aortic problems near term appear to be better managed by cesarean section followed promptly by treatment of the aortic disease. PMID- 3050158 TI - Tibial artery pseudoaneurysms: delayed complication of balloon catheter embolectomy. AB - Although complications of balloon catheter embolectomy are infrequent, the injury potential of these catheters is well recognized. This article describes a case of multiple tibial artery pseudoaneurysms that appeared 4 years after embolectomy in a 42-year-old man with otherwise normal arteries. The patient was treated by internal aneurysmorrhaphy without sequelae. A literature review of balloon catheter injuries yielded 46 cases categorized as arterial disruption (29), intimal injury (12), or catheter malfunction (5). These resulted in hemorrhage (13 cases), arteriovenous fistula (12), pseudoaneurysm (four), thrombosis (three), dissection (five), accelerated atherosclerosis (four), and catheter fragment embolism (five). Of these complications, only 41% were recognized during the initial operation. Direct observation detected 32% of these, whereas 68% were shown only by completion arteriography. Complications recognized during initial operation were more frequently asymptomatic without further surgery (84%) than those detected postoperatively (30%, p less than 0.001). Completion arteriography detected 87% of balloon catheter complications compared with only 23% of complications recognized intraoperatively without arteriography (p less than 0.001). We conclude that delicate technique, completion arteriography, prompt surgical treatment, and extended follow-up are important components of balloon catheter embolectomy. PMID- 3050159 TI - Cervical vessel injury after blunt trauma. AB - Blunt trauma accounts for 3% to 10% of cervical vessel injuries. Death and severe neurologic impairment have been reported in more than 80% of blunt carotid injuries. In our recent experience, 10 patients sustained 18 blunt cervical arterial injuries: two internal carotid artery (ICA) dissections, three ICA transections with pseudoaneurysm, five ICA thromboses, two vertebral artery dissections, one vertebral artery transection with pseudoaneurysm, one vertebral artery thrombosis, one minimal vertebral artery injury, and three caroticocavernous fistulas. A delay of more than 12 hours in making the diagnosis occurred in seven of the 10 patients. The mental status was initially normal in seven patients. The subsequent development of focal neurologic findings incongruent with CT scanning of the head prompted four-vessel angiography. Treatment was individualized and included supportive management, intravenous heparin, ligation, extracranial-intracranial bypass, and radiologic embolization. We have developed an angiographic classification that may aid management. Early angiography in patients with neurologic findings incongruent with head CT scan or in whom a normal sensorium and hemiparesis are present may permit improved outcomes. We advocate direct operative repair for accessible lesions of recent onset. For surgically inaccessible lesions, those with delayed presentation or in some cases with a fixed neurologic deficit, intravenous heparin can be started and follow-up angiography, head CT scanning, and the patient's clinical status determine further therapy. PMID- 3050160 TI - Prostacyclin and thromboxane A2 formation by atherosclerotic carotid artery: comparison with normal aorta, saphenous vein, and platelets. AB - Prostacyclin (PGI2) and thromboxane A2 (TxA2) formation by whole-tissue segments of nine carotid endarterectomy specimens (CES), five normal aortic specimens (NAS), six saphenous vein specimens (SVS), and four platelet samples were determined by incubation with 10 mumol/L 1-14C-radiolabeled prostaglandin endoperoxide H2 (PGH2), and in other experiments with and without 10 mumol/L of CGS 13080, a TxA2 synthase inhibitor. PGI2 formation (expressed as picomoles 6 keto-PGF1 alpha/2-min incubation per sample) by nonatheromatous proximal intima of CES (307 +/- 23, mean +/- standard error) and distal intima of CES (260 +/- 22) was not statistically different; however, it was greater than atheromatous transitional plaque (159 +/- 13 pmol) (p less than 0.01) and ulceration regions (140 +/- 15 pmol) (p less than 0.01) of CES, NAS (204 +/- 16 pmol) (p less than 0.01), and SVS (165 +/- 9 pmol) (p less than 0.01). TxA2 formation (expressed as picomoles TxB2/2-min incubation per sample) by CES ulceration (51 +/- 2 pmol) was low but greater than proximal (17 +/- 2 pmol) (p less than 0.01), distal (19 +/- 3 pmol) (p less than 0.01), and transitional (23 +/- 3 pmol) (p less than 0.01) regions. TxA2 formation by NAS and SVS was not detected (less than 10 pmol). CGS 13080 inhibited TxA2 formation by CES below the limits of detection. Incubation of 1.9 x 10(5) intact platelets with 10 mumol/L of PGH2 formed a quantity of TxA2 equal to that of CES ulceration.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3050162 TI - Transrectal ultrasonography: is it ready for routine use? PMID- 3050161 TI - Leads from the MMWR. Acute rheumatic fever among army trainees--Fort Leonard Wood, Missouri, 1987-1988. PMID- 3050163 TI - Assessing hospital-associated deaths from discharge data. The role of length of stay and comorbidities. AB - To assess the meaning of hospital-associated death rates, we studied whether mortality within 30 days of hospital admission (30-day mortality) is more informative than inpatient mortality and whether detailed assessment of additional discharge diagnoses helps in understanding death rates. We examined hospitalizations for elderly Medicare patients with principal diagnoses of stroke, bacterial pneumonia, myocardial infarction, and congestive heart failure; these conditions account for 30.8% of Medicare 30-day mortality. Average hospital stays for these conditions were 99.0% longer, and inpatient mortality was 25.0% higher in New York than in California, but 30-day mortality was 1.6% higher in California. We conclude that inpatient death rates depend on length-of-stay patterns and give a biased picture of mortality. Additional diagnoses such as shock and pneumonia were strongly associated with increased mortality, but Medicare data do not reveal which patients had these conditions at the time of admission. Recorded diagnoses of chronic diseases such as hypertension, diabetes mellitus, obesity, benign prostatic hypertrophy, and osteoarthritis were commonly associated with reduced risk of death; such reduced risk is not clinically plausible. Several lines of evidence suggest that chronic disorders are underreported for patients with life-threatening disorders. We recommend great caution in using discharge diagnoses of comorbid conditions to adjust hospital death rates for clinical differences in the patient populations. PMID- 3050164 TI - The African connection. Cotton Mather and the Boston smallpox epidemic of 1721 1722. AB - The contributions of black Americans to early American culture have still not been fully explored or given the attention deserved. A case in point is the significant contributions African slaves made to 18th-century American folk medicine. A review of the events incident to the smallpox epidemic in Boston in 1721 will illustrate the degree to which some reputable men of science depended on the testimony and experience of Africans in dealing with a particularly dread disease. PMID- 3050166 TI - Sixty years of neurological surgery. PMID- 3050165 TI - Overview of results of randomized clinical trials in heart disease. II. Unstable angina, heart failure, primary prevention with aspirin, and risk factor modification. PMID- 3050168 TI - Increased medication use in attention-deficit hyperactivity disorder: regressive or appropriate? PMID- 3050169 TI - Resource-based relative values. An overview. AB - Studies have been conducted over the past decade to develop a Resource-Based Relative Value Scale (RBRVS) for physicians' services. Policymakers view an RBRVS as a potential tool to pay physicians. The Physician Payment Review Commission, under a congressional mandate, has endorsed the general concept of a fee schedule based on resource costs for physician payment under Medicare. In this overview article, we present the policy context in which the RBRVS may play a role and describe the approach taken to develop this scale, specifically consultation with clinicians, researchers, and insurers and data gathering, including a national survey of physicians. We discuss underlying elements that are necessary to constructing an RBRVS, each of which is described more fully in subsequent articles: measuring the work (intraservice work) of performing medical services and procedures, estimating preservice and postservice work, comparing work across specialties, measuring practice costs, extrapolating from surveyed services, and establishing an RBRVS for evaluation/management services and for invasive procedures. Overall results are presented in a companion article. PMID- 3050167 TI - The alcohol-abusing patient: a challenge to the profession. PMID- 3050170 TI - Extrapolation of measures of work for surveyed services to other services. AB - A national survey of physicians produced detailed data on the work involved in performing 372 different services. This article describes methods developed to extrapolate the study data to a larger universe of services, defined by the Physicians' Current Procedural Terminology, edition 4. Because data measuring work inputs for nonsurveyed services presently are unavailable, we devised an extrapolation method that makes use of available charge data without building their inherent distortions into the extrapolated scale. To neutralize the effect of these distortions, we used small, homogeneous families of services as the basic units for the extrapolations and assumed that charges are reasonable indicators of relative work within such families. To produce extrapolated work values within each family, we multiplied an estimate of work based on survey data for a benchmark procedure by charge-based ratios that represent the relationships between surveyed and nonsurveyed services. These extrapolations can be used in constructing a Resource-Based Relative Value Scale. PMID- 3050171 TI - Results, potential effects, and implementation issues of the Resource-Based Relative Value Scale. AB - This article presents the overall results of the Resource-Based Relative Value Scale (RBRVS) study. We present resource-based relative values for selected services in each of the 18 specialties we studied. We found that preservice and postservice work represents close to 50% of total work for invasive services and 33% of total work for evaluation/management services. We also found that the work per unit time (a measure of intensity) for invasive services is about three times that of evaluation/management. We developed a simple model and simulated an RBRVS based fee schedule for the Medicare program under a "budget-neutral" assumption. Results for 30 commonly performed services show that office visit fees for evaluation/management services could rise by 70%, while some surgical fees could drop by 60%. We also simulated what the Medicare outlays would have been in 1986 for categories of medical services under an RBRVS-based fee schedule. We found that total Medicare payments for evaluation/management services would have increased by about 56%. Invasive, imaging, and laboratory services would have decreased by 42%, 30%, and 5%, respectively. We also discuss implementation issues related to an RBRVS-based fee schedule, such as the determination of a monetary conversion factor, practice costs, billing codes, and the need to evaluate the potential impacts of an RBRVS-based payment system on the cost and quality of health care. PMID- 3050172 TI - At last, a rational way to pay for physician's services? PMID- 3050173 TI - Physician payment reform: an idea whose time has come. PMID- 3050174 TI - The Resource-Based Relative Value Scale: a methodological and policy evaluation. PMID- 3050175 TI - From the Health Care Financing Administration. PMID- 3050176 TI - Leads from the MMWR. Transmission of HIV through bone transplantation: case report and public health recommendations. PMID- 3050177 TI - Suicide and cocaine. PMID- 3050178 TI - Captopril overdose and hypotension. PMID- 3050179 TI - Impact of a medical journal club on house-staff reading habits, knowledge, and critical appraisal skills. A randomized control trial. AB - The journal club is an established teaching modality in many house-staff training programs. To determine if a journal club improves house-staff reading habits, knowledge of epidemiology and biostatistics, and critical appraisal skills, we randomized 44 medical interns to receive either a journal club or a control seminar series. A test instrument developed by the Delphi method was administered before and after the interventions (mean, five journal club sessions). By self report, 86% of the house staff in the journal club group improved their reading habits vs 0% in the control group. Knowledge scores increased more in the journal club group than in the control group, and a trend was found toward more knowledge gained as more sessions were attended. Ability to appraise critically a test article increased slightly in each group, but there was no significant difference between the groups. We conclude that a journal club is a powerful motivator of critical house-staff reading behavior and can help teach epidemiology and biostatistics to physicians-in-training. PMID- 3050181 TI - Treatment of obesity in adults. Council on Scientific Affairs. AB - Concern with weight control should begin sufficiently early in life to reduce the risk of developing obesity. The complex etiology of obesity is, in part, responsible for the difficulty physicians encounter in treating this condition. Prevention is the "treatment" of choice. Early identification of individuals genetically at risk can be helpful in targeting those most likely to gain excess weight. Numerous dietary regimens have been devised in an attempt to achieve progressive weight loss in obese individuals. Since the ultimate goal of a weight reduction program is to lose weight and maintain the loss, a nutritionally balanced, low-energy diet that is applicable to the patient's life-style is most appropriate. Increasing energy expenditure through physical activity, in addition to decreasing energy intake, generally improves results in the management of obesity. Major changes in eating and exercise behaviors are necessary to ensure long-term weight control. Diet, exercise, and behavior modification are interdependent and mutually supportive. A comprehensive weight-reduction program that incorporates all three components is more likely to lead to long-term weight control. PMID- 3050180 TI - Orthotopic liver transplantation for alcoholic cirrhosis. AB - Fifteen patients with Laennec's cirrhosis underwent orthotopic liver transplantation between 1963 and the end of 1979. The first eight patients died perioperatively or within two months, but four of the next seven patients had long survival; three are still alive after 11 to 14 years. After the introduction of cyclosporine therapy, 41 more patients with alcoholic cirrhosis were treated with liver transplantation from 1980 to June 1987. The one-year survival is 73.2%, and, after one to three years, 28 (68%) of the recipients are living. Of the 35 patients in the combined old and new series who lived for six months or longer, only two returned to alcohol abuse. Social and vocational rehabilitation has been the rule in these recipients who were selected primarily because of urgency of need, because they or their families insisted on treatment, and because they and their families thereby committed themselves to long-standing programs of alcoholism care. PMID- 3050182 TI - [Current antimicrobial agent series II: Ticarcillin]. PMID- 3050183 TI - [Current antimicrobial agent series III: Norfloxacin]. PMID- 3050184 TI - [Studies on susceptibility of isolated organisms from pediatric infections against various antimicrobial agents]. AB - Twelve oral antimicrobial agents were tested for their antimicrobial activities against causative organisms isolated from pediatric infections. Activities of these antimicrobial agents against Streptococcus pyogenes were also examined in relation to T-antigen typing of the species. The results of the investigation are summarized as follows. 1. Against Staphylococcus aureus, rokitamycin (RKM), josamycin, ofloxacin, minocycline exhibited strong antimicrobial activities, and few strains of S. aureus showed resistance to these antimicrobial agents. More strains exhibited resistance to erythromycin (EM) than to other macrolide antibiotics (MLs) examined. Amoxicillin (AMPC)-resistance was often observed also. 2. Against S. pyogenes, beta-lactam antibiotics (beta-lactams) and RKM had MIC80 of 0.20 microgram/ml or below, and no resistant strains of this organism were observed against these antibiotics. Only 2 resistant strains (2.0%) of S. pyogenes to MLs were detected, but resistance to tetracyclines (TCs) was observed at a high frequency, with 71.4% or more strains among T-4, T-6, T-12 and T-28 antigen types exhibited resistance to TCs. Among the 21 strains of T-12 antigen type examined, only one strain (4.8%) was found resistant to MLs. These observations suggested that the reduction in the frequency of ML-resistant strains was not due to the reduction in the number of T-12 antigen type strains but due to losses of resistance factors against MLs of plasmids. 3. Antibacterial activities of beta-lactams and MLs against Streptococcus pneumoniae strains were good, similarly to activities against S. pyogenes. But many strains of S. pneumoniae were resistant to TCs. 4. New quinolone antimicrobial agents (quinolones) showed excellent activities against Branhamella catarrhalis strains with EM and RKM showing the next best activities. The number of resistant strains against quinolones, however, seemed to be on an increase. 5. Quinolones had strong antimicrobial activities against Haemophilus influenzae, few strains of which showed resistance to quinolones. AMPC had the next best activity, but approximately 10% of H. influenzae exhibited resistance to this antibiotic. 6. Against Campylobacter spp., quinolones and MLs showed the best activities with MIC80 values at or below 0.25 microgram/ml, and no resistant strains of the species against these antimicrobial agents were observed. Fosfomycin and TCs showed somewhat inferior activities to quinolones and MLs, with beta-lactams showing still lower activities. 7. Only few strains of Mycoplasma pneumoniae and Chlamydia trachomatis were examined, but MLs and TCs appeared to be effective against these organisms.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3050185 TI - [Laboratory and clinical studies of rokitamycin in pediatric fields]. AB - We have carried out laboratory and clinical studies on rokitamycin (RKM, TMS-19 Q). The results are summarized as follows. Serum and urinary concentrations of RKM were determined in 6 children with ages between 6 and 12 years given single oral doses of 5, 10 and 15 mg/kg. Mean serum concentrations peaked at 30 minutes after administration of 5, 10 and 15 mg/kg, and respective peak values were 0.30 microgram/ml, 0.79 microgram/ml and 1.32 micrograms/ml. Biological half-lives for 5, 10 and 15 mg/kg were 2.0 hours, 1.65 hours and 1.36 hours. The 6-hour urinary recovery ranged from 1.11% to 2.58% after administration of 5 mg/kg, and the mean 6-hour urinary recoveries were 1.35% after administration of 10 mg/kg and 2.28% after administration of 15 mg/kg. Therapeutic responses were recorded as excellent or good in 22 (73.3%) of the children, comprising 6 with tonsillitis, 2 with pharyngitis, 4 with bronchitis, 1 with bronchopneumonia, 1 with Mycoplasma pneumonia, 2 with whooping cough, 5 with streptococcal infections, 5 with Campylobacter enteritis, 3 with impetigo and 1 with SSSS. The microbiological effectiveness of RKM on identified pathogens comprising 4 strains of Staphylococcus aureus, 1 strain of Streptococcus pneumoniae, 6 strains of Streptococcus pyogenes, 4 strains of Haemophilus influenzae and 5 strains of Campylobacter spp. was not so satisfactory as evidenced by a eradication rate of 50.0%. No significant side effect due to the drug was observed in any cases. In conclusion, RKM was found to be efficacious and safe for the treatment of bacterial infections in children. PMID- 3050186 TI - [Microbiological, pharmacokinetic and clinical studies of rokitamycin dry syrup in the pediatric field]. AB - Rokitamycin (RKM), a newly developed macrolide antibiotic with a 16-membered ring, dissolves well under acidic conditions. It has been improved over other macrolides to minimize individual variations in its absorbability. We measured, using the GA-test, variations in gastric acidities of 43 children with ages between 1 to 14 years, and investigated the relationship between gastric acidities and pharmacokinetic values. Also activities (expressed in MICs) of antimicrobial agents were studied against clinically isolated 229 bacterial strains using an inoculum size of 10(6) cells/ml. Tested organisms included Streptococcus pyogenes (77 strains), Streptococcus agalactiae (29), Streptococcus pneumoniae (2), as Gram-positive cocci, and Haemophilus influenzae (1), Haemophilus parainfluenzae (1), Bordetella pertussis (12), Salmonella sp. (4) and Campylobacter jejuni (103) as Gram-negative bacilli. Against stock strains of bacteria, MICs of 10 drugs (RKM, erythromycin (EM), josamycin (JM), midecamycin (MDM), midecamycin acetate (MOM), clindamycin (CLDM), amoxicillin (AMPC), cefaclor (CCL), minocycline, ofloxacin (OFLX] were determined. Against isolates from patients who underwent treatment with RKM, MICs of only 4 drugs (RKM, EM, JM, MOM) were determined. Measurements were made on plasma and urinary concentrations of RKM and its urinary recovery rates after patients including 6 boys with ages between 5 years 1 month and 11 years 6 months were administered with RKM (dry syrup). Two groups of 6 boys were administered between meals with RKM at dose levels of 5 and 10 mg/kg, respectively. Clinical and bacteriological effects of RKM were evaluated for 175 patients including 5 cases of pharyngitis, 3 tonsillitis, 32 pneumonia, 17 mycoplasmal pneumonia, 34 atypical pneumonia, 28 streptococcal infections, 29 Campylobacter enteritis, 4 Salmonella gastroenteritis, and 23 enteritis due to unknown organisms. Five drop-out cases were excluded from the evaluations. In the evaluable cases, an average dose level used was 31.8 mg/kg/day, with a daily dose divided into 3 to 4 administrations and with an average treatment duration of 9 days. Adverse reactions of RKM and its effects on laboratory test values were investigated in these patients including the drop out cases. Obtained results of these studies are summarized below. 1. The GA-test produced pH values indicating that amounts of gastric acid were mostly either normal or high in 42 of the 43 subjects tested (97.7%), and only one low acid case (2.3%) was observed.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3050187 TI - [Pharmacokinetic, bacteriological and clinical studies of cefuzonam in the field of obstetrics and gynecology]. AB - Cefuzonam (CZON, L-105), an antibiotic injectable of cephalosporin family, was studied pharmacokinetically, clinically and bacteriologically to examine its distribution to female genital tissues and the activity on infections in the field of obstetrics and gynecology. Maximum concentrations in serum and genital tissues achieved 19-46 minutes after intravenous injection of CZON 1 g were 69.6 micrograms/ml for serum, 63.1 micrograms/g for oviduct, 34.2 micrograms/g for ovary, 22.5 micrograms/g for endometrium, 33.4 micrograms/g for myometrium, 30.7 micrograms/g for cervix uteri, and 37.1 micrograms/g for portio vaginalis. Clinical efficacies on 15 cases of intrauterine infection and adnexitis were proved with 4 cases of 'marked improvement' and 11 cases of 'improvement', thus the efficacy rate was 100%. Of 21 strains of aerobes and anaerobes isolated from infectious lesions, 19 strains were eliminated after administration of the drug. No side effects were observed. From these results of fundamental and clinical studies CZON appeared to be a highly useful drug fro the obstetric and gynecological infections. PMID- 3050188 TI - [Pharmacokinetic and clinical studies on the use of ceftizoxime in premature and newborn infants. The Chemotherapy Research Group for Mother and Child]. AB - A pharmacokinetic and clinical study of ceftizoxime (CZX), a newly developed cephem antibiotic intended for parenteral use, was conducted in premature and newborn infants, and resulted in the following findings. 1. Pharmacokinetics (1) Average serum half-lives (T 1/2) following a one shot intravenous dose of 20 mg/kg of CZX to mature and premature newborn infants in age groups of 0-3, 4-7, 8 14, and 15-30 days were: 4.14, 3.01, 2.57, and 1.98 hours (for mature infants) and 5.26, 4.59, 3.71, and 2.64 hours (for premature infants), respectively, decreasing with ages in days of the infants. (2) Average T 1/2's after a one shot 10 mg/kg dose was similar to that after a 20 mg/kg dose. Serum concentrations of CZX were dose-dependent. (3) T 1/2 after 1-hour intravenous drip infusion revealed a same trend after one shot injection. (4) Urinary in the first approximately 6 hours recoveries following a 20 mg/kg one shot dose to mature and premature newborn infants were as follows: 35% (0-3 days old) and 45-55% (4 days old and older) in mature infants and 30% (0-3 days old) and 45% (4 days old and older) in premature infants. 2. Therapeutic effectiveness (1) The subjects examined were 112 newborn infants consisting of 83 with infections and 29 who received CZX for prophylaxis. The 83 infants had 86 cases of infections, which were classified as A, when the causative organisms were identified and as B, when the causative organisms were not identified. Rates of therapeutic effectiveness were 95.0% for group A and 95.7% for group B. Bacteriological effectiveness were studied on 41 strains isolated from group A, and were as high as 89.5% for Gram positive organisms and 95.5% for Gram-negative organisms. The rate of successful prophylaxis for the 29 infants was 96.6%. (2) Side effects did not occur among the 120 newborn infants. Laboratory tests with abnormalities included leukopenia, neutropenia, eosinophilia, thrombocytosis and increased GOT or GPT. These pharmacodynamic and clinical findings have fully substantiated the satisfactory therapeutic usefulness of CZX in both the treatment and prevention of neonatal infections in the usual dose of 20 mg/kg, which is to be given b.i.d. for infants up to 3 days old; b.i.d. or t.i.d. for infants 4 to 7 days old, and t.i.d. or q.i.d. for infants 8 days old and older. The drug can be given in a daily dose as high as 120 mg/kg when infection is severe. PMID- 3050189 TI - [Clinical efficacy and pharmacokinetic evaluation of ceftizoxime in neonates and young infants]. AB - Thirteen neonates and young infants, including 5 infants with very low birth weight, were treated with ceftizoxime (CZX) and its clinical efficacy and side effects were evaluated. The ages of the patients ranged from 0 to 96 days, and their body weights ranged from 580 to 5,050 g. Doses given were 20-54 mg/kg every 6 to 12 hours for 2.5 to 7.5 days. Two infants with sepsis, one with urinary tract infection, one with sepsis and urinary tract infection, and 1 with fetal infection were considered to have responded satisfactorily to the CZX treatment. The drug was well tolerated and side effects was not apparent. Pharmacokinetic studies were done on CZX in 8 patients including 4 infants with very low birth weight. Their ages ranged from 2 to 91 days, and body weights from 545 to 5,050 g. Serum concentrations at 2 hours after single 20 mg/kg intravenous bolus injections were 19.2 to 44.2 micrograms/ml and the levels were 2.11 to 26.3 micrograms/ml at 8 hours. Elimination half-lives of CZX ranged 1.90 to 9.57 hours in these patients. In 2 infants with very low birth weights with ages 7 and 91 days, half-lives were as long as 9.57 and 8.24 hours, respectively. Urinary recovery in 6 hours was 31.9-66.9% in 5 patients. Urine concentrations of the drug in 24 samples collected at various time from the 7 patients ranged from 130 to 3,219 micrograms/ml. Influence of CZX on the fecal flora was studied in 1 patient given 20 mg/kg X 4/day of the drug.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3050191 TI - [Pharmacokinetic, bacteriological and clinical studies of ceftizoxime in neonates]. AB - Pharmacokinetic, bacteriological and clinical studies of ceftizoxime (CZX) were performed in neonates. 1. Serum concentrations and urinary excretion of CZX were investigated in 12 neonates ranging ages from 1 to 27 days (gestational age, 35 41 weeks; birth weight, 2,150-4,030 g) and 2 infants ranging ages from 55 to 57 days (gestational age, 39-40 weeks; birth weight, 2,320-2,650 g). Each of the subjects was given a single intravenous dose of 20 mg/kg by one shot. Serum concentrations of CZX in the neonates were 24.9-53.7 micrograms/ml at 1/4 hour after intravenous injection, with an average of 40.6 +/- 7.6 micrograms/ml. Serum half-lives of CZX were 1.32-4.75 hours and averaged 2.60 +/- 1.06 hours. Serum concentrations ranged from 2.01 to 14.6 micrograms/ml at 6 hours after injection with an average of 7.70 +/- 3.89 micrograms/ml. In the 2 infants, serum concentrations were 42.0 and 46.2 micrograms/ml at 1/4 hour (average: 44.1 +/- 3.0 micrograms/ml), and 2.91 and 5.04 micrograms/ml at 6 hours after injection (average: 3.98 +/- 1.51 micrograms/ml). Half-lives were 1.54 hours in 1 infant and 1.93 hours in the other (average: 1.74 +/- 0.28 hours). Furthermore, 6-hour urinary recovery rates were 28.5-71.7% (average: 49.3 +/- 12.8%) in the neonates and 42.1-55.5% (average: 48.8 +/- 9.5%) in the infants. The above results suggest that the following 3 points are accepted; 1) peak serum concentrations (at 1/4 hour) in neonates were similar to those in infants and older children irrespective of age (days after birth). 2) Serum half-lives of CZX in neonates shortly after birth were 4 or 5 times longer than those in older children, but decreased rapidly with the advance of day-ages. The half-life in neonates of 2 weeks of age or so became shorter to about twice the normal value in infants. Furthermore, half-lives of the drug in those at an age of the first half of infancy were similar to those in older children. 3) The urinary excretion rates tended to be somewhat low with neonates soon after birth, but became very similar to those in infants and older children at a relatively early stage.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3050192 TI - [Pharmacokinetic, bacteriological and clinical studies of ceftizoxime in neonates and low birth weight infants]. AB - Ceftizoxime (CZX), one of the fifth group in cephem-antibiotics classified by Fujii, was administrated intravenously with a one shot dose of 20 mg/kg to neonates and low birth weight infants with ages ranged 4-21 day old and plasma and urinary concentrations and urinary recovery rates of the drug were determined. Clinical, prophylactic and bacteriological effects of CZX were evaluated and adverse reactions and effects on laboratory test values due to this drug were studied in 22 neonates and low birth weight infants (0-76 day old) consisting of 16 cases with various bacterial infections including presumptive cases of bacterial infections and 6 cases with prophylactic administration against infectious diseases. An average CZX daily dose of 55.3 mg/kg was given once or divided into 2-4 times daily (twice: 16 cases, 3 times: 3 cases, 4 times: 2 cases) through intravenous one shot administration for 6 days on the average. The results obtained are summarized as follows. 1. Neonates and low birth weight infants were classified into 3 groups by age: 4-7, 8-14, 15-21 day old. Plasma peak levels of CZX were observed at an average of 5 minutes after administration in all 3 groups with mean values of 58.3, 74.9 and 76.9 micrograms/ml, respectively, and the 4-7 day old-group showed a lower value than with other 2 groups. Mean values of AUC were 218.9, 221.0 and 197.0 micrograms.hr/ml, respectively, and no notable difference was observed within each group. Mean values of half-lives of CZX were 3.61, 2.72 and 2.37 hours, respectively, and the younger group tended to have the longer value. 2. Urinary concentrations of CZX ranged 10.9-1.190 micrograms/ml in all of the 3 groups during 0-2, 2-4, 4-6, 6-8 hours after administration. Mean values of urinary recovery rates during 8 hours of the 3 age-groups were 60.1, 68.7, 56.7%, respectively. The oldest group showed the lowest mean value because one of the cases had the lowest value of 34.5% for an unknown reason. 3. Clinical effect of CZX in 16 cases with various bacterial infections and presumptive bacterial infections was evaluated with an efficacy rate of 87.5%. The prophylactic effect was recognized in all 6 cases that were given CZX to prevent infectious diseases. 4. The bacteriological effect was evaluated in only one case with an infection due to Escherichia coli, which was eradicated by the treatment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3050194 TI - [Studies of ceftizoxime in perinatal period]. AB - Bacteriological studies and clinical evaluations of ceftizoxime (CZX) in perinatal period were carried out, and results are summarised as follows: Antibacterial activities of CZX in amniotic fluid were determined using the broth dilution method, and bactericidal effect on Streptococcus agalactiae, Staphylococcus aureus and Escherichia coli were demonstrated. The bactericidal effect of CZX increased in amniotic fluid and remarkable increases of activities against resistant strains were demonstrated. The penetration of CZX into mother's milk was low, and it was speculated that the drug transfer to the newborn through breast feeding was very little. Clinically, CZX was effective in the treatment of perinatal infections without any side effect. The above results has demonstrated that CZX is a clinically useful antibiotic for the prophylaxis and the treatment of perinatal infections. PMID- 3050190 TI - [Pharmacokinetics and clinical studies of ceftizoxime in newborn and premature infants]. AB - Serum concentrations and urinary recovery rates of ceftizoxime (CZX) were investigated in 13 mature newborns and 16 premature newborns (with ages 1-17 days) after one shot intravenous injection of 10 and 20 mg/kg, respectively, for treatment and prophylaxis of various infections. Obtained data were studied comparatively among the 3 groups, i.e. the 1st group (age: 0-3 days), 2nd group (age: 4-7 days) and 3rd group (age: 8 days or older). The clinical investigation was made in 9 male and 6 female newborns aged 3-34 days including 4 pediatrics patients with septicemia (1 with purulent meningitis, 1 with urinary tract infection, 1 with Staphylococcal pneumonia and 1 without complication), 1 with maxillary sinusitis, 5 with bronchopneumonia and 5 with urinary tract infections. 1. Serum concentrations and urinary recovery rates (1) Mature newborns given one shot intravenous injection of 10 mg/kg Serum concentrations of the drug in the 1st, 2nd and 3rd groups in 30 minutes after one shot intravenous injection of 10 mg/kg peaked at 18.9-23.3 micrograms/ml, without any significant differences, and thereafter gradually declined to 2.10-4.99 micrograms/ml at 8 hours. Serum half lives were shorter in older subjects and values in the 3 groups were 3.89, 2.99 and 2.35 hours, respectively. Urinary recovery rates ranged from 55.5% to 70.0% at 8-6 hours in the 3 patients. (2) Mature newborns given one shot intravenous injection of 20 mg/kg Serum concentrations of the drug in the 3 groups peaked in 30 minutes after one shot intravenous injection of 20 mg/kg at, respectively, 36.9, 41.4 and 38.4 micrograms/ml, and thereafter they gradually declined to 9.0, 7.3 and 4.5 micrograms/ml at 8 hours, respectively. Serum half-lives were shorter in older subjects and values in the 3 groups were 3.59, 2.93 and 2.49 hours, respectively. Urinary recovery rates ranged from 43.5 to 78.2% within 12 hours in 2 patients, within 8 hours in 2 patients and within 6 hours in 1 patient. (3) Premature newborns given one shot intravenous injection of 10 mg/kg Serum concentrations of the drug in the 3 groups peaked in 30 minutes after one shot intravenous injection of 10 mg/kg at, respectively, 27.9, 21.5 and 23.0 micrograms/ml, and they gradually declined thereafter to 10.8, 6.2 and 6.5 micrograms/ml at 8 hours, respectively. Serum half-lives were shorter in older subjects and values in the 3 groups were 5.28, 4.43 and 4.24 hours, respectively.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3050193 TI - [Pharmacokinetic and clinical studies of ceftizoxime in the perinatal period. The Chemotherapy Research Group for Mothers and Children]. AB - Pharmacokinetic and clinical studies of ceftizoxime (CZX) in the perinatal period gave the following results: 1. Peak concentrations of CZX in the maternal serum, umbilical cord serum and amniotic fluid in mothers after one intravenous injection of 1 g were, respectively, 70.2 micrograms/ml at 0 hour; 15.7 micrograms/ml at 0.5 hour; and 10-30 micrograms/ml at 3-6 hours. Concentrations of CZX in the neonatal serum were 0.87-13.5 micrograms/ml during 6-14 hours after parturition. The mean concentration of CZX in the milk in 1-8 hours after injection was less than 0.32-0.52 microgram/ml. 2. Good or excellent clinical efficacy was obtained in 28 of the 29 patients with perinatal infections, with an efficacy rate of 96.6%. Prophylactic effectiveness was obtained in 14 of the 15 patients, with an efficacy rate of 93.3%. 3. No side effects were observed in 44 cases. GOT and GPT values increased slightly in 1 patient. No abnormal values in total serum bilirubin or other parameters were found in any neonates after parturition. 4. The above results suggest that CZX is safe and effective for the treatment and prophylaxis of infection in the perinatal period. PMID- 3050195 TI - [Pharmacokinetic, bacteriological and clinical studies of cefuzonam in the field of obstetrics and gynecology. Study group of cefuzonam in the field of obstetrics and gynecological infections]. AB - A multi-center open study was conducted to investigate cefuzonam (CZON, L-105) regarding to its pharmacokinetic, bacteriological and clinical aspects in the field of obstetrics and gynecology with the participation of 31 medical institutions and the related facilities. The results are summarized as follows. 1. Peak MICs of CZON for Staphylococcus aureus, coagulase (-) staphylococci, Escherichia coli, Klebsiella pneumoniae, Bacteroides fragilis group, Peptostreptococcus spp. isolated from obstetrical and gynecological infections with relatively high frequencies were 0.39, 0.20, 0.024, 0.024-0.05, 12.5, 0.20 microgram/ml, respectively, with an inoculum size of 10(6) CFU/ml. 2. When 1 g of CZON was given through bolus injection, the maximum concentration (Cmax) of CZON in pelvic dead space exudate was 18.7 micrograms/ml at 60.9 minutes (Tmax) after the injection; Cmax's in all female genital tissues were observed at 0.6-27.9 minutes and ranged from 11.9-26.3 micrograms/g. The Cmax 8.3 micrograms/ml, in the pelvic dead space exudate was noted at 97.0 minutes after the end of the intravenous drip infusion of 1 g over 1 hour, and Cmax's in genital tissues were 14.3-30.0 micrograms/g at the end of infusion. With 1 hour drip infusion of 2 g, Cmax's in genital tissues were 35.0-53.9 micrograms/g at the end of infusion. 3. The clinical efficacy of CZON was evaluated in 206 evaluable patients with obstetric and gynecologic infections. Efficacy rates classified by types of infections were 97.1% (67/69) for intrauterine infections, 81.6% (31/38) for intrapelvic infections, 91.8% (45/49) for adnexitis, 95.2% (20/21) for infections of the external genital organs and 86.2% (25/29) for other infections. 4. Side effects were observed in 7 of the 262 patients: eruption in 6 cases, itching in 2, diarrhea in 1. Abnormal laboratory test values were noted in 9 of the 256 patients. Most of them were slight elevation of hepatic function values. CZON showed satisfactory clinical efficacy and potent antibacterial activity, hence it appears that CZON will be a very useful antibiotic for obstetric and gynecologic infections. PMID- 3050196 TI - Intracellular sodium concentration in lymphocytes and erythrocytes of patients with essential hypertension. AB - In order to investigate the abnormalities in intracellular sodium concentration ([Na]i) in essential hypertension, [Na]i in lymphocytes and erythrocytes was determined in 37 outpatients with essential hypertension and 35 normotensive controls matched for sex and age. [Na]i in lymphocytes was higher in hypertensive patients than in normotensive controls, while no difference was observed in [Na]i in erythrocytes between the 2 groups. There was no significant correlation between [Na]i in lymphocytes and erythrocytes in either group. In hypertensive patients, plasma renin activity had a negative correlation with [Na]i in lymphocytes, but not with [Na]i in erythrocytes. These results suggest that the increase in [Na]i may be associated with essential hypertension with suppressed plasma renin activity and that lymphocytes are more suitable than erythrocytes for the analysis of [Na]i. PMID- 3050197 TI - Periodic fluctuation of blood pressure and its management in a patient with pheochromocytoma. Case report and review of the literature. AB - The case of a 69 year old man with a right adrenal pheochromocytoma who manifested cyclic fluctuations of blood pressure with a cycle length of 9 to 13 min is reported. We collected and reviewed 14 similar cases previously reported in the literature. In these cases, right adrenal pheochromocytoma was most common, while 1 case involved the left adrenal and 2 cases were of extra-adrenal origin. The incidence in males was twice that in females and the median age was 45.3 years. Although a good correlation between the blood pressure and plasma norepinephrine concentrations was observed in our patient, the exact mechanism for the cyclic fluctuations of blood pressure is not known. In our patient, both YM-09538 and bunazosin were effective in controlling severe hypertension preoperatively. YM-09538 induced significant increases in urinary norepinephrine concentrations (1327 +/- 238 micrograms/day), while bunazosin induced a significant decrement in urinary norepinephrine concentrations (475 +/- 188 micrograms/day) compared with pretreatment levels (900 +/- 42 micrograms/day) (p less than 0.01). These observations indicated that bunazosin as a postsynaptic alpha-adrenergic receptor blocker interfered with release of norepinephrine from the tumor and thus might be beneficial in the management of elevated blood pressure in patients with pheochromocytoma. PMID- 3050198 TI - [Oncogenes and differential diagnosis among non-neoplastic, precancerous lesions and early and advances cancers]. AB - Reviewing recent publications, especially on ras and myc expression in GI tract tumors, the usefulness of detecting oncogene changes for differential diagnosis among non-neoplastic changes, precancerous lesions, early and advanced cancers were discussed. While quantitative change of oncogene expression per se appears to have only limited value, qualitative alteration in oncogenes or in regulatory mechanisms of oncogene expression seems to offer more promising perspective for judging the biological character of individual lesions. The definition of precancerous lesion and early carcinoma is also carefully discussed. PMID- 3050199 TI - [Early cancer in the head and neck]. AB - In head and neck tumors, there is no definition of early cancer. If T1N0M0, no lymph node and distant metastasis, will be early cancer, it is difficult to detect the tumor at early phase in head and neck tumor except for tongue and laryngeal cancer. Because it is hard to define it at early stage in the tumor surrounding by bony structure or tumor growing endophytically. Although almost tumor of head and neck were easily diagnosed by inspection and palpation, total imaging examination should be done actively for detection of early stage of cancer. PMID- 3050200 TI - [Clinical characteristics and imaging diagnosis of early cancer in the hepatobiliary and pancreatic region]. AB - For the present, we don't have any reliable definition of the early cancer in hepato-biliary and pancreatic region. In this paper three examples of comparatively early cancer were taken up; (1) among hepatocellular carcinoma, so called, small liver cancer; (2) among gallbladder carcinoma and bile duct carcinoma, those whose extent of infiltration within pm and fm respectively; (3) among pancreatic carcinoma, those whose maximum diameter are 2 cm. And the system of imaging diagnosis was discussed as well as the discussion of their clinical characteristics, imaging diagnosis and the differential diagnosis. PMID- 3050201 TI - [Diagnostic imaging of small liver tumors]. AB - The recent introduction of the various imaging modalities including MRI has improved the diagnostic accuracy of metastatic liver tumor. Especially ultrasonography provides useful informations to demonstrate the relation of tumors to surrounding structures. However, at the same time it is still a fact that small lesions frequently escape detection and qualitative diagnosis is also rendered impossible in cases of small liver tumors. Repeated ultrasonographic examination combined with clinical informations is most important and qualitative diagnosis seems accomplishable by a combination of various imaging techniques including ultrasonically guided biopsy. PMID- 3050202 TI - [A follicular, medium-sized cell lymphoma showing minimal progress in the last 13 years--case report]. AB - A case history of a 39-year-old woman with a follicular, medium-size cell type lymphoma (Nodular poorly differentiated lymphocytic lymphoma) is reported. In spite of receiving no treatment, her lymphoma had changed little for 13 years. This case is regarded as a good example for considering the treatment of low grade malignant lymphomas. PMID- 3050205 TI - [Non-RI DNA probes]. PMID- 3050206 TI - [Medicolegal documental materials chronologically arranged in the Meiji era (Suppl.)]. PMID- 3050204 TI - [Metabolic disorders--from biochemistry to DNA diagnosis]. PMID- 3050207 TI - Plasma volume in nephrotic patients showing chronic glomerulonephritis with normal kidney function. PMID- 3050203 TI - [A case of cystadenocarcinoma of the liver complicated by gastric cancer--a review of 83 cases of primary cystic liver cancer in the literature]. AB - So far as is known, this is the first reported case in the literature of a primary cystic liver cancer complicated with a gastric cancer. A 77-year-old female was admitted to hospital because of a swelling in the upper abdomen and appetite loss. CT scanning, an echogram, and an angiogram of the abdomen revealed a large cystic lesion which originated from the left lobe of the liver. A DIC indicated a gall stone, Further, upper GI fluoroscopy and endoscopy revealed a gastric cancer of the Borrmann II type, and thus a dome resection of the cyst, cholecystectomy, aad 3/4 partial gastrectomy was performed. The liver was found to contain much mucin. The histology indicated a cystadenocarcinoma of the liver, and an adenocarcinoma of the stomach of a poorly differentiated type. PMID- 3050208 TI - Effects of captopril and nicardipine on renal hyperfiltration and hypertrophy in uninephrectomized diabetic rats. PMID- 3050209 TI - Effects of thromboxane A2 synthesis inhibition on renal renin release and protein excretion in diabetic rats. PMID- 3050210 TI - [A clinical study on edema formation in nephrotic patients]. PMID- 3050211 TI - [Influence of sodium intake on antihypertensive effect of calcium antagonist]. PMID- 3050212 TI - Molecular approaches to learning and memory. AB - We are now at the stage of neurophysiology where learning and memory can be subjects of studies at a strictly molecular level, on the basis of the well established finding that these higher nervous activities are sustained by, and formed in, the physicochemical events of specified neural mechanisms in the brain. As for the neurophysiological process of memory, much evidence has shown that short-term memory and long-term memory probably result from different molecular events in the brain, i.e., the former from reversible chemical modification of the synapses concerned, and the latter from reorganization of the synapses following synthesis of protein and its axonal transport, which causes the enduring consolidation of memories. How does the experience of individual organisms trigger the protein synthesis in the brain required for long-term memory? What is the role of protein molecules thus formed? What is the mechanism for the regulation of gene expression in the reorganization of neuronal circuits? Many such difficult problems need to be solved. Recently, cholinergic and glutamatergic neuron networks have attracted much attention because there is a strong possibility that they play a critical role in memory. The clinical implication of these findings in human memory deficit, as exemplified in senile dementia, further emphasizes the importance of neurobiological elucidation of the molecular mechanism for learning and memory. PMID- 3050213 TI - Ruggero Oddi. To commemorate the centennial of his original article--"Di una speciale disposizione a sfintere allo sbocco del coledoco". PMID- 3050214 TI - Extravascular lung water with special reference to thoracotomy, manual lung manipulation and rapid fluid transfusion. AB - An increase in extravascular lung water (EVLW) is indicative of pulmonary edema and the measurement of EVLW has been considered as an important clue for the early detection of pulmonary edema, which is a serious complication of lung surgery. The influence of thoracotomy, manual lung manipulation and rapid fluid transfusion on EVLW were experimentally studied in the lung tissue of dogs, using the double indicator dilution method and comparing it with the drying method. The following results were obtained. 1) EVLW measured by the double indicator dilution method correlated well with EVLW measured by the drying method of tissue at 80 degrees C for 48 hours. 2) EVLW was significantly increased immediately after rapid fluid transfusion. EVLW increased as the osmotic pressure in the transfused fluid became higher. 3) Even the simple procedure of thoracotomy resulted in an increase of EVLW and the addition of manual compression to the lung facilitated this increase of EVLW even further. PMID- 3050215 TI - Emergency versus elective cholecystectomy in acute cholecystitis. AB - The present study was carried out at the Department of Surgery, All India Institute of Medical Sciences Hospital, New Delhi, between January 1982 and October 1984. Clinical diagnosis of acute cholecystitis was confirmed by ultrasound scanning or Tc99m labelled HIDA Scan. Group I (n=24) comprised patients who underwent emergency cholecystectomy while Group II (n=23) comprised patients who were managed conservatively, and had an elective cholecystectomy performed 6-12 weeks later. There was no mortality or wound infection in either group. The incidence of other complications was 8.3 per cent and 8.6 per cent in Groups I and II, respectively. Emergency cholecystectomy (Group I) reduced the total hospital stay of a patient by approximately 70 per cent and post cholecystectomy syndrome was seen in Group II patients only. We thus recommend emergency or early cholecystectomy during the acute stage of cholecystitis, as it is safe, effective and economical, provided it is done by an experienced surgeon. PMID- 3050216 TI - Changes of glycolytic enzyme activities and plasma insulin levels in diabetic herbivorous voles and KK mice. PMID- 3050217 TI - Demonstration of Toxoplasma gondii antigen in stillborn piglets using immunoperoxidase technique. PMID- 3050219 TI - [Macular edema in diabetic retinopathy]. PMID- 3050218 TI - [Diabetic retinopathy. Classification and spontaneous development]. PMID- 3050220 TI - [Effect of intensive treatment on diabetic retinopathy]. PMID- 3050222 TI - [Regulation of genetic expression of the enzymes of glycolysis and gluconeogenesis in the liver of the diabetic]. PMID- 3050223 TI - [Insulin resistance: a target]. PMID- 3050221 TI - [Hypoglycemic risk in IDDM]. PMID- 3050224 TI - [Prospective studies on recent and long-term diabetic neuropathy]. PMID- 3050225 TI - [Physiopathology of atherosclerosis in the diabetic]. PMID- 3050226 TI - [Diabetes and coronaropathy]. PMID- 3050227 TI - [Coronary insufficiency in the diabetic]. PMID- 3050228 TI - [Non-invasive exploration of arteritis in the diabetic. Therapeutic applications]. PMID- 3050229 TI - [The 1988 Apollinaire Bouchardat prize. The role of lipid substrates in adaptation to fasting in the newborn infant and child]. PMID- 3050230 TI - [The 1988 Maurice Derot prize. A century of research on the ideal supplementary insulin therapy (1889-1988)]. PMID- 3050231 TI - [Physiopathology of non-insulin-dependent diabetes. New therapeutic approaches with gliclazide]. PMID- 3050232 TI - [Action of metformin on vascular risk factors in diabetes]. PMID- 3050233 TI - [What are we learning about spontaneous insulin-dependent diabetes in laboratory animals?]. PMID- 3050234 TI - [Value of animal models for understanding human obesity]. PMID- 3050235 TI - [Choice of therapeutic modality for type II diabetes]. PMID- 3050236 TI - [Physiopathology of insulin resistance, obesity and type II diabetes in animals]. PMID- 3050237 TI - [A diabetic under intensive insulin therapy]. PMID- 3050238 TI - [Pregnancy and non-insulin-dependent diabetes. Taking charge and managing a type II pregnant diabetic woman, or one who would like to become pregnant. Attitude toward a diabetic discovered during pregnancy]. PMID- 3050239 TI - [Arterial hypertension in the diabetic: support of most of its complications]. PMID- 3050240 TI - The Kansas Medical Society. Membership Directory 1988. PMID- 3050241 TI - Acute myelogenous leukemia: problems in the elderly patient. PMID- 3050242 TI - Immunology of the human gastrointestinal tract in health and disease--a brief review. PMID- 3050243 TI - [Infections due to Mycobacterium ulcerans and Mycobacterium haemophilum]. PMID- 3050244 TI - Using storytelling to teach relaxation to children. PMID- 3050246 TI - Fifth annual John Peters award, American Society of Nephrology. Award recipients: Jacob Churg and Conrad Pirani. PMID- 3050245 TI - Identification of glomerular immune deposits in cryoglobulinemia glomerulonephritis. AB - To provide further evidence of the nature of intraglomerular immune deposits in essential mixed cryoglobulinemia (EMC), we used two mouse monoclonal antibodies against cross-reactive idiotypes present on monoclonal rheumatoid factors (MoRFs) from patients with type II-EMC. MoAb Cc1 reacted with 9 of 16 circulating IgMk MoRFs tested, and MOAb Lc1 with four of the remaining. Using indirect immunofluorescence and immunoperoxidase techniques, we could identify the same cross-reactive idiotype of the serum MoRF in the renal biopsy specimens from 11 of 13 patients with EMC glomerulonephritis. Kidney specimens from the three patients, whose MoRF was not recognized by MoAbs Cc1 and Lc1, were negative. Two out of 30 control renal biopsies from patients with other forms of glomerulonephritis were shown to contain idiotype (Cc1 and Lc1) positive material. Both patients had serum polyclonal RF which could account for this finding. In conclusion, our results provide direct evidence that serum cryo-MoRF participate in the formation of glomerular immune deposits and, presumably, in the pathogenesis of renal damage in EMC glomerulonephritis. PMID- 3050247 TI - Cytoskeleton organization and submembranous interactions in intestinal and renal brush borders. PMID- 3050248 TI - Confirmation of the utility of fine needle aspiration biopsy of the renal allograft. AB - Allograft immunobiologic theory would predict that analysis of immunocompetent cells infiltrating the renal transplant would be most instructive. Recently a new aspiration biopsy technique has been developed to permit such analysis in patients which can be safely and repetitively performed. The clinical utility of such a technique has been tested utilizing a randomized prospective trial in which an aspirate was obtained every other day from the third post-operative day until discharge. Analysis included examination of adequacy criteria and the capacity of pathologic diagnosis to corroborate clinical diagnosis from coded specimens. Ninety-six aspirates from 21 consenting transplant recipients were obtained and analyzed. In 94 instances a clinical diagnosis could be made; 80 aspirates fulfilled adequacy criteria. We found the technique to be highly sensitive (greater than or equal to 90%) and highly specific (greater than or equal to 90%) for the clinical diagnoses of acute allograft rejection, post operative acute renal failure, cyclosporine toxicity, and normal function. We conclude that the fine needle aspiration technique is an important adjunct to analysis of clinical renal transplantation and offers a major advantage to the clinical scholar in understanding transplant biology. PMID- 3050251 TI - [Reye syndrome]. PMID- 3050250 TI - Distinguishing minimal-change disease from mesangial disorders. PMID- 3050252 TI - [Bone marrow transplantation in childhood. II]. PMID- 3050249 TI - Creatinine measurements often yielded false estimates of progression in chronic renal failure. AB - In 9 of 22 observation periods (lasting an average of 15 months) in 17 patients with moderate to severe chronic renal failure (GFR 4 to 23 ml/min), rates of progression as estimated from the linear regression on time of 24-hour creatinine clearance (b1) differed significantly from rates of progression as estimated from the regression on time of urinary clearance of 99mTc-DTPA (b2), during all or part of the period of observation. b1 exceeded b2 in four cases and was less than b2 in the other five. Thus there were gradual changes in the fractional tubular secretion of creatinine in individual patients, in both directions. Owing to these changes, measurements of creatinine clearance gave erroneous impressions of the rate or existence of progression during all or a portion of the period of observation in nearly half of these patients. In the 22 studies as a group, using the entire periods of observation, b1 indicated significantly more rapid progression (by 0.18 +/- 0.06 ml/min/month, P less than 0.01) than did b2, and had a significantly greater variance. Measurements of progression based on the rate of change of reciprocal plasma creatinine (multiplied by an average rate of urinary creatinine excretion in each study) were equally misleading, even though less variable. We conclude that sequential creatinine measurements are often misleading as measures of progression and should, when feasible, be replaced by urinary clearance of isotopes in following patients with chronic renal failure. PMID- 3050253 TI - [Ethics in pediatrics. Observations on the problem of ethical decision making support in care-giving]. PMID- 3050255 TI - [Secretory immunoglobulin A (SIGA) in the saliva of newborn infants]. PMID- 3050256 TI - [Clinical contribution to the sonographic diagnosis and perinatal treatment of fetal ovarian cysts]. PMID- 3050257 TI - [Anterograde pyelography under ultrasonic control]. PMID- 3050254 TI - [Is there a green light for the use of green light in phototherapy of neonatal icterus?]. PMID- 3050259 TI - [The place of duodenogastric reflux in the genesis of diseases of the unoperated and operated stomach]. PMID- 3050260 TI - [A case of forgotten foreign body in the abdominal cavity perforating the intestinal wall and entering the intestinal tract]. PMID- 3050261 TI - [Treatment of protruding ears]. PMID- 3050258 TI - [Percutaneous nephrostomy under ultrasonic control]. PMID- 3050262 TI - [A new method for anastomosing the colon to the rectum following Hartmann's operation]. PMID- 3050263 TI - [Cholelithiasis in the neonatal period]. AB - We report about a four weeks old male infant who developed jaundice with cholangitis and fever caused by cholelithiasis which could be confirmed by sonographic examination. Familiar history revealed several relatives with gallstones. PMID- 3050264 TI - [Principles for evaluating simple, diagnostic tests]. AB - The fundamental terms in the evaluation of diagnostic tests are described and the differences to corresponding test parameters in clinical chemistry are emphasized. The derivation of a two-by two contingency table is explained in detail and basic characteristics of diagnostic tests, i.e. sensitivity and specificity as well as their independence of the prevalence (in the sense of a priori-probability) is pointed out. The importance of the so-called predictive values is shown graphically, also the problem of selecting a cut-off-level (discrimination value) for the evaluation of quantitative data. The various attempts to find a single value for the description of a diagnostic test are discussed and it is stressed that there is no principal necessity to use other terms than those mentioned above. Nevertheless it is possible to give a prevalence-independent measure which has some additional advantages in rating and comparing simple diagnostic tests. PMID- 3050265 TI - Role of diuretics, hormonal derangements, and clinical setting of hyponatremia in medical patients. AB - Because hyponatremia is frequently associated with preceding diuretic treatment and unrestricted fluid intake--conditions which have not been addressed sufficiently in published literature--we studied the pathophysiology and the clinical setting of such hyponatremia in a large group of internal medicine patients. We observed: a) Of an initial 310 patients with chemical hyponatremia only 204 (64%) had an associated plasma hypoosmolality. Since a normal plasma osmolality excludes a disturbance of water metabolism only the 204 patients with hypoosmolar hyponatremia were included in the study. This data shows that plasma osmolality is an essential measurement in any evaluation of hyponatremia. b) In 204 consecutive patients with hypoosmolar hyponatremia the electrolyte disturbance was related to advanced congestive cardiac failure in 25%, decompensated liver cirrhosis in 18%, volume contraction in 28%, syndrome of inappropriate antidiuretic hormone secretion in 19% and renal insufficiency in 4%. c) Plasma vasopressin was measurable in 90% of the 204 patients. It is known that radioimmunoassays to measure vasopressin fail to reliably detect low concentrations of circulating vasopressin (less than 0.5 pg/ml). It may therefore be stated that hypoosmolar hyponatremia was generally characterized by a failure of antidiuretic hormone suppression. d) Mean daily fluid intake of hyponatremic patients was 2.35 +/- 0.15 l. In the presence of stimulated vasopressin this large a fluid intake is bound to worsen the severity of hyponatremia. e) Of 204 patients 126 were treated with diuretics at the time of study. In these patients hyponatremia worsened during such treatments and was associated with evidence of prerenal azotemia. However there were no significant differences between diuretic treated and -untreated patients with respect to plasma vasopressin stimulation and amount of fluid intake.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3050266 TI - Apo E 2 phenotypes in type II diabetics with and without insulin therapy. AB - In 106 type II diabetics with persisting hyperlipidemia (i.e. persistently increased triglycerides greater than 200 mg/dl during intensive diabetes therapy) the Apo E polymorphism was examined in relation to IDDM (n = 68) and NIDDM (n = 38). It was shown that Apo E2 phenotypes are more (22.6% vs. 14.5%, p less than 0.05) and Apo E3 phenotypes less frequent in type II diabetics than in non diabetic controls (86.8% vs. 94.7%, p less than 0.001). Looking at the increase in Apo E2 phenotypes it could be proved that the phenotype composition was distinctly different between diabetics with and in those without insulin therapy. While in NIDDM the increase was consequent to a higher concentration of Apo E2 homozygotes (p less than 0.005) it was caused by Apo E2 heterozygotes in IDDM (p less than 0.025) accompanied by a simultaneous decrease in Apo E3 homozygotes (p less than 0.025). Regarding blood lipids there was an increase in total cholesterol due only to VLDL cholesterol in IDDM as well as in NIDDM. It is concluded that in spite of similar hyperlipidemias in type II diabetics the increase in Apo E2 phenotypes is different; it is induced by heterozygotes in IDDM and by homozygotes in NIDDM. PMID- 3050269 TI - [A study to develop a play therapy program for children who need hospitalization and surgery--literature review]. PMID- 3050268 TI - The effect of low and high NaCl diets on oral glucose tolerance. AB - The effects of low and high NaCl diets on plasma glucose and insulin responses to glucose ingestion were investigated in 15 patients with essential hypertension. Oral glucose (75 g) tolerance tests were carried out while patients were taking diets with low (2 g/day) and high (20 g/day) NaCl content. Fasting plasma glucose and insulin levels were both significantly lower during ingestion of the high NaCl diet (p less than 0.05). After glucose ingestion, the incremental areas under the two hour plasma glucose and insulin curves were significantly smaller during ingestion of the high NaCl diet (glucose p less than 0.005 and insulin p less than 0.025). These findings that low NaCl diets increase the glycemic response to glucose loads suggest that use of NaCl restriction for the treatment of essential hypertension may not always be desirable. PMID- 3050267 TI - Very low density lipoprotein apolipoprotein B metabolism in humans. AB - The human plasma lipoproteins encompass a broad spectrum of particles of widely varying physical and chemical properties whose metabolism is directed by their protein components. Apolipoprotein B100 (apo B100) is the major structural protein resident in particles within the Svedberg flotation range 0-400. The largest of these, the very low density lipoprotein (VLDL), rich in triglyceride, are metabolised by sequential delipidation through a transient intermediate density lipoprotein (IDL) to cholesterol-rich low density lipoproteins (LDL). Several components contribute to the regulation of this process, including (a) the lipolytic enzymes lipoprotein lipase and hepatic lipase (b), apolipoproteins B, CII, CIII and E, and (c) the apolipoprotein B/E or LDL receptor. Lipoprotein lipase acts primarily on large VLDL of Sf 60-400. Hepatic lipase on the other hand seems to be critical for the conversion of smaller particles (Sf 12-60) to LDL (Sf 0-12). Although most apo B100 flux is directed to the production of the delipidation end product LDL, along the length of the cascade there is potential for direct removal of particles from the system, probably via the actions of cell membrane receptors. This alternative pathway is particularly evident in hypertriglyceridaemic subjects, in whom the delipidation process is retarded. VLDL metabolism shows inter subject variability even in normal individuals. In this regard, apolipoprotein E plays an important role. Normolipidaemic individuals homozygous for the apo E2 variant exhibit gross disturbances in the transit of B protein through the VLDL-IDL-LDL chain. PMID- 3050270 TI - Hazards of urethane (ethyl carbamate): a review of the literature. AB - Urethane (ethyl carbamate) is used alone or in combination with other drugs to produce anaesthesia in laboratory animals. Although originally studied as a potential phytocide, urethane demonstrated antineoplastic properties when administered to rats with the Walker rat carcinoma 256. Subsequent trials in humans led to its use as a chemotherapeutic agent for various leukaemias. Mice develop pulmonary adenomas earlier in life and at a higher incidence following urethane administration. Urethane's carcinogenic influence is greater in neonatal mice; it also has a transplacental influence in mice. In rats, urethane increases the incidence of pulmonary adenomas, Zymbal Gland tumours, and a variety of other neoplasms. Urethane is absorbed sufficiently from the skin of laboratory animals to produce a transient narcosis. The carcinogenic effect appears to be due to an undefined oncogenic intermediate formed in the blood. Considering the properties urethane demonstrates in animals, the safety of its use by laboratory personnel is in question. However, if appropriate guidelines are followed, urethane should continue to be a useful anaesthetic agent for laboratory animals. PMID- 3050271 TI - von Willebrand factor and the pathophysiology of thrombotic thrombocytopenia: from human studies to a new animal model. PMID- 3050272 TI - Morphology and biochemistry of bone remodeling: possible control by vitamin D, parathyroid hormone, and other substances. AB - The effects of PTH and vitamin D on bone are the result of their direct and indirect effects on the functional cells of bone remodeling units and their precursors. These effects are probably modified or controlled by growth factors, cytokines, and PGs generated locally by the process of bone remodeling. Bone remodeling includes resorptive and bone forming phases, each with a longitudinal and a radial component of progression in time and space. Longitudinal resorption is rapid, prolonged and is probably carried out by osteoclasts utilizing hydrogen ions and lysosomal enzymes to remove mineral and organic components of bone in a highly localized and directed fashion. Individual osteoclasts are probably long lived cells with a nuclear and perhaps a cytoplasmic turnover rate of 8%/day, with replenishment coming from preosteoclasts in the reversal zone. Radial resorption is slower and shorter than longitudinal resorption. It is carried out by reversal phase monocytes whose exact relationship to osteoclasts is not clear. Activated collagenase diffusing from osteogenic cells in the reversal zone could also play a role. The longitudinal rate of bone formation is probably a measure of the rate of proliferation and differentiation of osteogenic cells at the site at which they were activated. The radial rate of bone formation is a measure of how rapidly osteoblasts synthesize and mineralize bone matrix once they reach the resorption surface. PTH and vitamin D have no direct effects on mature osteoclasts. They may have direct stimulatory effects on proliferation and differentiation of osteoclast precursors and their fusion with osteoclasts but this is not clear because the ontogeny of osteoclasts vis a vis monocytes and other phagocytic cells is still not clear. It is likely that their effects to increase osteoclast precursors involve interactions among lymphocytes, monocytes, and hematopoietic stem cells at a distance from bone remodeling units and are mediated by 1,25(OH)2 vitamin D3 induced synthesis of cytokines and colony stimulating factors. Stimulatory effects of PTH, vitamin D, PGs, and cytokines on osteoclasts are mediated by as yet undefined factors produced by osteoblasts. Osteoblasts stimulated by PTH could also inhibit osteoclasts by synthesizing and releasing PGs. PTH and vitamin D have diverse and often contradictory effects on the functional activity of osteoblast-like cells in vitro that are difficult to interpret because the relationship of these cells to osteoblasts in vivo is not clear.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3050273 TI - Complement and leukocyte independent proteinuria and eicosanoid synthesis in rat membranous nephropathy. AB - We examined the effect of complement depletion and leukocyte depletion on an experimental model of membranous nephropathy. Nephrosis was induced in 200-gm male Sprague-Dawley rats by priming with cationic bovine gamma-globulin in adjuvant on day 1 followed by intravenous challenge with antigen starting on day 10. No naive control rats had immunofluorescent deposits in glomeruli; urine protein was less than 10 mg/24 hour and glomerular thromboxane synthesis was 658 +/- 64 ng/mg glomerular dry weight. In contrast, all rats primed and challenged with cationic bovine gamma-globulin had intense granular capillary wall deposits of rats IgG, bovine gamma-globulin, C3 and C5; severe proteinuria (183 +/- 24 mg/24 hour) was observed, associated with a 3-fold increase in glomerular thromboxane (2,393 +/- 574 ng/mg, all p less than 0.01 versus naive controls). In some rats, administration of cobra venom factor and antiserum to rat C3, starting on day 8 was used to deplete complement; hemolytic C3 and C5 were less than 2% of normal at sacrifice. These rats had IgG and bovine gamma-globulin deposits, whereas they lacked glomerular C3 or C5. Proteinuria (209 +/- 28 mg/24 hour) and glomerular thromboxane (2,087 +/- 394 ng/mg) were markedly increased compared with control, but no different from normocomplementemic rats primed and challenged with cationic bovine gamma-globulin. In other rats, depletion of leukocytes was achieved by 1,000 R x-irradiation on day 7; at sacrifice, irradiated rats had 1,270 +/- 462 wbc/microliter versus 10,375 +/- 1,652 in nephrotic rats given no other treatment, with unaltered differentials. These rats had glomerular deposits of rat IgG, bovine gamma-globulin, C3 and C5 indistinguishable from nephrotic rats with normal leukocyte counts in intensity and distribution. Proteinuria (202 +/- 30) and glomerular thromboxane (2,358 +/- 189 ng/mg) were markedly elevated compared with naive controls, but were not different from the normocomplementemic or complement-depleted groups primed and challenged with antigen. An additional control group included rats primed with ovalbumin on day 1, irradiated with 1,000 R on day 7, and challenged with cationic bovine gamma-globulin starting on day 10. This group had granular capillary wall deposits of bovine gamma-globulin, but not deposits of IgG, C3, or C5; urine protein excretion (less than 10 mg/24 hours) and glomerular thromboxane synthesis (613 +/- 90) were not different from naive controls. Glomerular prostaglandin E2 synthesis did not differ among the five groups.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3050274 TI - Antiglomerular basement membrane nephritis in the mouse. Study on the role of complement in the heterologous phase. AB - The role of complement was examined in a model of antiglomerular basement membrane nephritis in the mouse, induced by intravenous injection of goat anti mouse glomerular basement membrane serum, and characterized by early glomerular lesions and a dose-dependent albuminuria. We compared the reaction after injection of the antiserum in complement-normal B10.D2 new mice with that in congenic, congenitally C5-deficient B10.D2 old mice, and in both strains after C3 depletion by treatment with cobra venom factor. The dose-dependent albuminuria was not affected by the absence of complement activation. Also, deficiency of C3 and/or C5 had no inhibitory effect on the histologic glomerular lesions: it did not reduce the influx of polymorphonuclear granulocytes in the glomerular capillary vessels, nor prevented the eventual intravascular coagulation. We conclude that complement-independent mechanisms are involved in the development of the heterologous phase of antiglomerular basement membrane nephritis in the mouse. PMID- 3050275 TI - "The dean" Robert Wilson, M.D. 1867-1946. PMID- 3050276 TI - Health effects on workers in the pharmaceutical industry: a review. PMID- 3050277 TI - Solutions A and B: cyanide antidote or folklore myth? PMID- 3050279 TI - Quantitative determination of nuclear estrogen receptors by an enzyme immunoassay: applicability and caveats. AB - A method is presented with which approx. 95% of nuclear estrogen receptors appear to be extracted from MCF-7 cells. Since both nuclear isolation and nuclear estrogen receptor extraction take place in a single test tube with only vortex mixing, loss of nuclear material is minimized. The amount of nuclear estrogen receptors in the nuclear extract was determined by direct [3H]estradiol labeling of monolayer cultures and with a commercially available estrogen receptor immunoassay (ER-EIA) kit. Since the ER-EIA kit was designed and calibrated for quantitative determination of cytosolic estrogen receptor isolated in low ionic strength buffer, the applicability of the ER-EIA to quantitative determination of estrogen receptor content in high ionic strength nuclear extraction buffer was tested. A linear relationship exists between the amount of nuclear estrogen receptor detected by the immunoassay, the amount of receptor present in serial dilutions of the nuclear extract and the amount of nuclear estrogen receptor detected in cells by [3H]estradiol labeling of monolayer cultures, the absolute amount of nuclear estrogen receptors determined by the immunoassay consistently exceeded the amount of receptor detected by [3H]estradiol labeling. The possibility that the enzyme immunoassay must be properly calibrated for the specific conditions of the nuclear estrogen receptor assay is discussed. PMID- 3050278 TI - Differential inhibition of estrogen and antiestrogen binding to the estrogen receptor by diethylpyrocarbonate. AB - Diethylpyrocarbonate differentially inhibited the specific binding, in lamb uterine cytosol, of estradiol (inhibition approximately 90% with 4 mM reagent) and 4-hydroxytamoxifen (inhibition approximately less than 50% with 4-16 mM reagent), a potent triphenylethylene antiestrogen. Saturation analysis experiments indicated that the effects of diethylpyrocarbonate were due to progressive but differing decreases in the concentration of binding sites for the two ligands, with no apparent change in the affinity constants. However, competitive binding and dissociation experiments evidenced that steroidal and nonsteroidal estrogens still bound, but with very low affinities, to diethylpyrocarbonate-modified receptor (greater than 1000-fold decrease in affinity) whereas the affinities of triphenylethylene antiestrogens were much less affected (less than 10-fold decrease). Both ligands prevented the inactivation of the estrogen receptor by diethylpyrocarbonate, estradiol being more efficient than 4-hydroxytamoxifen. These data indicate that the action of diethylpyrocarbonate results in the formation of two populations of estrogen receptor that are quantitatively nearly equivalent: the first does not bind estrogens or antiestrogens; the second does not bind estrogens significantly but still interacts with antiestrogens at a high affinity. The simplest interpretation is that these two populations arise from mutually exclusive modifications by diethylpyrocarbonate of at least two aminoacid residues located at or close to the ligand binding site; modification of one residue totally prevents the binding of estrogens and antiestrogens; the modification of the second impairs only the binding of estrogens. Considering that (i) hydroxylamine, which specifically reverses the diethylpyrocarbonate-induced modification of histidine and tyrosine residues, restored a large part (greater than 80%) of the estradiol- and 4-hydroxytamoxifen-binding capacity of diethylpyrocarbonate inactivated cytosol, and that (ii) similar differential inhibition of estrogen and antiestrogen binding was observed following the action of tetranitromethane, it is likely that these residues are histidine(s) and/or tyrosine(s). These results evince a marked difference in the interaction of estrogens and triphenylethylene antiestrogens with the estrogen receptor, which could account for the altered activation of the receptor by triphenylethylene antiestrogens. Consequently, the screening of ligands with modified steroid receptors could be a useful method for distinguishing between potential hormone agonists and antagonists. PMID- 3050280 TI - Alcohol and drug problems in the schools: results of a national survey of school administrators. AB - A mail survey of public high-school administrators in the United States was conducted to gather information about the nature and extent of school problems with student alcohol and drug use. Of the 728 schools selected for the national metropolitan probability sample, 543 (75%) returned a completed questionnaire. The results indicated that from 1980 to 1985, about one in six students attended schools that reported a serious problem with student alcohol use. In contrast, the proportion of students attending schools with a serious drug problem decreased from about one in four in 1980-81 to about one in seven in 1984-85. The most common explanation provided for a decreasing student alcohol or drug problem was changes in the school's discipline policy or increased enforcement of the existing policy. Few respondents attributed a reduction in student alcohol or drug problems to prevention or treatment programs. Limitations of the survey data are discussed as well as directions for future research. PMID- 3050281 TI - Does alcohol advertising affect overall consumption? A review of empirical studies. AB - This article reviews the empirical warrant for the assertion that alcohol advertising affects overall alcohol consumption. Econometric, exposure, experimental studies and advertising bans are examined. The evidence indicates that advertising bans do not reduce alcohol sales, total advertising expenditures have no reliable correlation with sales of alcoholic beverages, and that experimental studies typically show no effect of advertising on actual consumption. However, one set of studies does show that drinkers are exposed to more television alcohol advertisements, without making the causal connection clear. In general, the evidence indicates little impact of alcohol advertising on alcohol sales or drinking. However, some results are suggestive and there is a need for more sophisticated econometric, exposure and experimental studies that take into account a wider range of variables. PMID- 3050283 TI - Incidental detection of carcinoma of the prostate while staging carcinoma of the rectum with transrectal ultrasound. AB - This case report describes the unanticipated results of a transrectal ultrasound performed to stage a cancer of the rectum. During sonography, the normally dense prostate peripheral zone produced weak echoes indicating the existence of an unexpected carcinoma of the prostate. The presence of extensive prostate cancer was subsequently verified by biopsy. PMID- 3050282 TI - Immunohistological characterization of human monoclonal antibody against lung cancer. AB - Using immunoperoxidase staining method, a human monoclonal antibody (MAb), termed HB4C5, which was generated by fusion of NAT-30 cells with regional lymph node lymphocytes from a lung cancer patient, was examined for tumor specificity. The determinant recognized by HB4C5 MAb was well preserved in usual formalin-fixed paraffin-embedded tissue sections. Experiments with four major histological types of the lung cancer showed that HB4C5 MAb reacted with adenocarcinoma (19/27), squamous cell carcinoma (2/15), and large cell carcinoma (4/6), but not with small cell carcinoma (0/7). The immunoreaction was seen in the cytoplasm. In addition, this MAb also reacted with some gastric and colon adenocarcinomas. Further study on normal lung and nonpulmonary tissues revealed that HB4C5 MAb bound only type II pneumocytes and proximal tubules of normal kidney. PMID- 3050285 TI - Should the temperature chart influence management in cardiac operations? Result of a prospective study in 314 patients. AB - The body temperature is measured routinely and carefully charted in our own and presumably all units. Pyrexia is normal after bypass and is discounted on the basis of clinical experience in the first few days. If this pyrexia persists, a search for infection may be instigated and discharge from the hospital may be delayed. A clinical trial of antibiotic prophylaxis provided the opportunity to collect and collate 6-hourly temperature observations for 314 patients for 1 week after operation. The length of bypass and the presence of lower respiratory tract infection were positively correlated with the duration of postoperative fever. However, neither surgical sepsis nor urinary tract infection had any consistent effect on the duration or magnitude of postoperative fever in the first week. PMID- 3050284 TI - In memoriam Julian Johnson (1906-1987). PMID- 3050286 TI - A new model for assessment of lung preservation. AB - No completely satisfactory experimental model exists to compare different techniques of preservation currently used in the distant procurement of a lung allograft. A canine model of left lung transplantation is described in which an inflatable cuff is placed around each pulmonary artery. Each cuff is connected to a subcutaneous reservoir, which allows alternate occlusion of either pulmonary artery. Functional assessment of lung function is made during ventilation of both lungs and after a 10-minute period of perfusion to the native lung alone and then to the transplanted lung alone. Systemic and pulmonary artery pressures are recorded continuously, and measurement of arterial blood gases and oxygen uptake are made immediately after the operation and again at 3 days. The animal is then put to death and the lungs are excised and weighed. Five dogs underwent transplantation of the donor lung immediately after excision (mean ischemic time = 55 +/- 7 minutes). Similar values for oxygen tension and oxygen uptake were obtained postoperatively for the right lung (oxygen tension = 420 mm Hg, oxygen uptake = 101 ml/min) and the left lung (oxygen tension = 368 mm Hg, oxygen uptake = 108 ml/min). However, carbon dioxide tension was elevated (right lung = 41 mm Hg, left lung = 52 mm Hg). Mean pulmonary artery pressure increased during allograft perfusion (right lung = 14 mm Hg, left lung = 24 mm Hg), although systemic blood pressure was unchanged. Similar results were observed at 3 days. The mean weight of the native lung was 101 +/- 2 gm and that of the transplanted lung, 128 +/- 6 gm. This model achieves consistent survival and allows serial observations of the functional adequacy of an allograft compared with a normal contralateral lung. PMID- 3050287 TI - Zipper sternotomy: a new approach to an old problem. PMID- 3050288 TI - Autologous bone marrow transplantation in acute leukaemia. PMID- 3050289 TI - The effect of 13-cis retinoic acid on hematopoiesis in human long-term bone marrow culture. AB - The modulatory effect of 13-cis retinoic acid (RA) on the growth, differentiation and function of hematopoietic cells in human long-term cultures was studied. RA (5 X 10(-8) M) induced enhancement of myeloid progenitor cell growth in the non adherent layer throughout 6 weeks of incubation while it did not affect the number of myeloid progenitors in the adherent layer. The vitamin did not alter the differentiation pattern of colony forming unit-culture (CFU-C). The addition of RA to cultures for 5 weeks did not alter the cellular composition of the adherent layer. Prolonged exposure of hematopoietic cells to RA did not affect the functional activity of neutrophils and macrophages, i.e. the cells were active in phagocytosing Candida albicans (CA). PMID- 3050290 TI - Cytogenetic studies on rat thymic lymphomas induced by N-propyl-N-nitrosourea. AB - N-Propyl-N-nitrosourea (PNU) was proved to be a strong leukemogen, which induces myelogenous leukemia or thymic lymphoma in rats. BUF/Mna rats and F344 rats were the strain most susceptible to thymic lymphomagenic activity of PNU. In addition, F1 rats between BUF/Mna and WKY rats were also susceptible to PNU-lymphomagenic activity. In the present experiment, karyotypes of 31 thymic lymphomas induced by PNU in BUF/Mna rats and in F1 rats between BUF/Mna and WKY rats were analysed for chromosomal abnormalities. Although no specific chromosomal abnormalities were observed throughout all lymphomas, del(11q) and dup(2q) were observed frequently in BUF/Mna rat lymphomas. Breakpoints and/or fusion-points were frequently observed in chromosome 11, followed by chromosomes 2, 5 and 6. Trisomy of chromosome 7, on which c-myc oncogene is mapped, was observed in seven cases, and monosomy of chromosomes 12, 18, 19, 20 and X was seen in seven or eight cases each, though these changes were generally observed in minor cell population in each case. PMID- 3050291 TI - [Ultrasonic diagnosis in the prevention of shock in placenta praevia]. PMID- 3050292 TI - Granular lymphocyte proliferative disorders: report of 12 cases and review of the literature. PMID- 3050294 TI - Evidence for pluripotent stem cell origin of idiopathic myelofibrosis: clonal analysis of a case characterized by a N-ras gene mutation. AB - Three cases of idiopathic myelofibrosis were screened for the presence of mutations at codon 12, 13, or 61 of the ras gene family by a rapid method based on polymerase chain reaction and hybridization to mutation-specific oligonucleotides. PB cells of one patient showed a point mutation at codon 12 of the N-ras oncogene. This molecular genetic hallmark was used to investigate the clonal relationship of different cell lineages by cell separation analysis. Presence of the N-ras 12 mutation in granulocytes, monocytes, B cells, and T lymphocytes, as well as erythroblasts, indicates that idiopathic myelofibrosis originates from a pluripotent stem cell, at least in this patient. PMID- 3050295 TI - Lack of differential sensitivity of normal hematopoietic stem cells and murine lymphoblastic leukemic cells to a lysosomotropic agent, N-dodecyl morpholine. AB - The effect of N-dodecyl morpholine (NDM), a lysosomotropic compound, on the clonogenic capacity of GK15, Sp2.0, Hb131, and L1210 lymphoblastic tumor cells and CFU-GM and CFU-S progenitor cells from DBA/2 mice was measured in order to evaluate the potential use of this compound for the purging of tumor-contaminated bone marrow (BM) in autologous BM transplantation. The growth of clonogenic tumor cells from all of the tested cell lines was inhibited with doses of NDM that also killed 100% CFU-GM and CFU-S, and no optimal dose could be found in this animal model to purge marrow while sparing sufficient stem cells to ensure engraftment in syngeneic BM transplantation. PMID- 3050296 TI - New and evolving concepts in leukemia. PMID- 3050297 TI - Mayo administration and Harry J. Harwick. PMID- 3050293 TI - Clinical evaluation of a DNA probe assay for the Philadelphia (Ph1) translocation in chronic myelogenous leukemia. AB - We report the clinical evaluation of an improved DNA probe assay for the characteristic genetic marker of human CML, observed by cytogenetics and designated the Philadelphia chromosome (Ph1). The Ph1 chromosome results from the fusion of c-abl proto-oncogene sequences from chromosome 9 to phl gene sequence on chromosome 22. (The phl gene is often referred to as bcr. However, for clarity we prefer to reserve the designation "bcr" for the region within the phl gene in which translocation breakpoints have been found to occur. We also find it useful to distinguish between two such regions in phl, bcr-210 and bcr-190, named after the 210- and 190-kDa phl/abl fusion proteins resulting from translocations with breakpoints in the respective regions. We refer to the corresponding chromosomal translocations as Ph1(bcr-210) and Ph1(bcr-190).) DNA, extracted from peripheral blood (PB) or bone marrow (BM) and digested with restriction endonuclease BglII, is hybridized with a probe (phl/bcr-3) spanning a breakpoint cluster region within phl. Rearrangements are revealed by the presence of one or two novel junction fragments. Clinical specimens from leukemic patients with active disease were compared by cytogenetic and DNA probe analysis at seven centers in the United States and Europe. The probe assay identified the phl rearrangement in 190 of 191 cases of Ph1-positive CML, as well as in 12 of 27 clinically diagnosed CML specimens lacking a typical Ph1 chromosome. DNA rearrangements also were seen in two of six cases of Ph1-positive ALL. No false positive results were obtained among 93 non-leukemic controls. Mixing experiments showed that the DNA probe assay can detect as few as 1% leukemic cells in a specimen. A preliminary study of CML patients in remission after allogeneic BM transplantation revealed a small fraction of residual Ph1-positive leukemic cells in a significant number of such patients. PMID- 3050298 TI - Statistical considerations for performing multiple tests in a single experiment. 4. Performing multiple statistical tests on the same data. PMID- 3050299 TI - Paul Dudley White: pioneer American cardiologist. PMID- 3050300 TI - Treatment of Parkinson's disease with pergolide: a double-blind study. AB - Pergolide is a potent dopamine agonist and is known to have anti-Parkinson properties. We administered pergolide to patients with suboptimal control of Parkinson's disease who had a short-duration response to carbidopa-levodopa in a 6-month, double-blind study. Pergolide added to the carbidopa-levodopa regimen resulted in both subjective and objective improvement in comparison with placebo. In patients who tolerated pergolide, the median time spent in the "off" (parkinsonian) state was reduced from 5.0 to 2.2 hours daily (compared with a 0.3 hour reduction in the placebo group). These patients were able to decrease the median frequency of carbidopa-levodopa dosage from 7.5 to 5.0 doses daily (no change in the placebo group). Prolongation of the "on" response (optimal response to treatment) to single doses of drugs was corroborated by monitoring of the patients' Parkinson response cycle. The peak response was also improved in most patients. Of 25 patients randomized to the pergolide group, 7 were unable to tolerate this drug; confusion or hallucinations occurred in 4 of these patients, and chest pain, leukopenia, and nonspecific dizziness, respectively, developed in the other 3. All adverse events were reversible with reduction of the dose or discontinuation of the pergolide regimen. In conclusion, patients with Parkinson's disease who experience clinical fluctuations with carbidopa-levodopa may be helped by the addition of pergolide to the therapeutic regimen. PMID- 3050302 TI - Use of health services by black children according to payment mechanism. AB - The use patterns of approximately 2,600 black children, categorized according to type of insurance (Medicaid, private health insurance or no insurance), were analyzed. All children were enrolled in an urban pediatric primary care program that attempted to increase access to health care by poor children. Medicaid recipients used health-care services more than their counterparts who had private or no insurance. All groups received significant levels of preventive care. The percentage of health care received in the emergency room did not vary significantly among the groups. These results suggest that special delivery systems can be effective in reaching poor children and eliminating usage differentials according to income. PMID- 3050301 TI - Pergolide: long-term use in Parkinson's disease. AB - Previous short-term studies have shown that the dopamine agonist pergolide improves control of Parkinson's disease when used in conjunction with carbidopa levodopa (Sinemet). We assessed the long-term outcome (2 1/2- to 3-year follow up) in patients with Parkinson's disease who participated in our previous pergolide double-blind trial and were subsequently switched to open-label pergolide therapy. Of 41 evaluable patients who began pergolide therapy, 10 (24%) experienced sustained substantial benefit that persisted to the end of this investigation. A total of 23 patients (56%) remained on pergolide therapy and, as a group, had considerable improvement over baseline at 2 1/2 to 3 years on the basis of several measurements of efficacy. A tendency toward deterioration could be reversed in many patients by larger or more frequent doses of carbidopa levodopa; nevertheless, all but four patients were still taking the same dose or less of carbidopa-levodopa at the end of this study as at the onset. Patients with the best initial response to pergolide seemed most likely to experience long term benefit. Confusion and hallucinations were the side effects most likely to necessitate discontinuation of pergolide. Symptoms suggestive of dose-related angina pectoris occurred in four patients in the open-label phase and two patients in the earlier double-blind phase (13% of patients who started pergolide therapy); these symptoms were easily controlled by dose reduction or discontinuation of pergolide, without sequelae. Dose-related leukopenia developed in one patient. PMID- 3050303 TI - [Cyclosporin A in cadaveric kidney transplantation. A randomized study]. PMID- 3050304 TI - [Toxicologic research in Spain as seen through the databases (1985-1986)]. PMID- 3050305 TI - [Does an idiopathic hematuria really exist?]. PMID- 3050306 TI - [Autotransfusion]. PMID- 3050307 TI - [CA 125, a marker of ovarian epithelial tumors]. PMID- 3050308 TI - [Oxalate-induced arthropathy in hemodialysis]. PMID- 3050310 TI - [Therapeutic perspectives in acquired immunodeficiency syndrome]. PMID- 3050309 TI - [Rhombencephalitis caused by Listeria monocytogenes]. PMID- 3050311 TI - [Cocaine: the epidemic to come]. PMID- 3050312 TI - [Dawn phenomenon. Studies using artificial pancreas in insulin dependent diabetic patients]. PMID- 3050314 TI - [Perinephritic abscess]. PMID- 3050313 TI - [Clinico-epidemiological study of 57 cases of Klebsiella bacteremia]. PMID- 3050315 TI - [Problems in the care of patients with acquired immunodeficiency syndrome]. PMID- 3050317 TI - [Non-Hodgkin's lymphomas: importance of prognostic factors in therapeutic decisions]. PMID- 3050316 TI - [Long-term treatment of arterial hypertension in diabetics with captopril]. PMID- 3050318 TI - [Is gout correctly diagnosed?]. PMID- 3050319 TI - [The efficacy of diagnostic tests (II)]. PMID- 3050320 TI - [Subclinical renal tubular dysfunction in systemic lupus erythematosus]. PMID- 3050321 TI - [Changes in the renin-aldosterone axis in systemic lupus erythematosus]. PMID- 3050323 TI - [Solitary splenic abscess treated by splenotomy]. PMID- 3050322 TI - [Streptococcus agalactiae bacteremia in adults: study of 46 cases admitted to the Hospital Vall d'Hebron (1978-1985)]. PMID- 3050324 TI - [Disseminated pigmented nevus syndrome]. AB - Two patients carriers of a neuro-cutaneous syndrome showing scattered pigmentary nevus and neurologic disorders are exposed, who are added to three similar cases presented in a previous publication. Differences with other syndromes that show cafe-au-lait spots, like those of Recklinghausen and Albright, and also of other known publications, are remarked. Ultrastructural studies are contributed. The denomination of "disseminated pigmentary nevus syndrome" is proposed in comparison with the epidermal nevus syndrome. PMID- 3050326 TI - [Immunology of tertiary pinta]. AB - We've studied the immunological performed of twenty two natives Tikunas suffering from tertiary pinta. Among those, patients had been treated previously (two years earlier) which 2,400,000 IU of G benzathine penicillin, and twelve had no treatment. Both groups demonstrated an increment in the IgM synthesis (72.72%), IgG (50%), indicating the presence of strong antigenic stimuli. The great majority presenting a negative response revealed also a reduction in the cellular immune competence to at least two of the tests performed (92.3%), when were realized the PPD, DNCB and skin grafts tests. PMID- 3050325 TI - [Skin involvement in Hodgkin's disease]. AB - We report a case of a previously diagnosed patient of Hodgkin's disease, nodular sclerosis type and IVLDB stage. This patient subsequently presented a dermatologic nodule. A specimen cutaneous biopsy was performed and the histopathologic study demonstrated a dermal infiltrate with many classic Reed Sternberg giant cells. These findings were consistent with the skin involvement in Hodgkin's disease. Specific cutaneous lesions are rare in this lymphoma and therefore it may be argued that our patient had a lymphomatoid papulosis. However, the protracted clinical course would not support this concept, because the dermatologic lesion responded to chemotherapy and it is well-known that the clinical course of the lymphomatoid papulosis is unaffected by these treatments. We concluded that our patient is a case of specific skin involvement in Hodgkin's disease. PMID- 3050327 TI - [The amyloidoses]. AB - There are many clinical types of amyloidosis, as many as chemical types of amyloid substance would be its substratum. This clinic-chemical plurality permits us to talk about the amyloidosis. This article is specially guided to those amyloidosis that present cutaneous damages during its clinical course. They are the immunocytic amyloidosis and the keratinocytic amyloidosis. PMID- 3050328 TI - [Risk factors for the recurrence of spinocellular epithelioma. Study of 650 cases]. AB - An evaluation is made of the clinical and histological parameters of 650 cases of spinocellular epithelioma. The statistical analysis shows tumor relapse to be related to tumor size, evolution time, relapse following previous treatments, earlier cutaneous lesions, Broders' grading, acantholysis, inflammatory infiltrate and type of treatment applied. PMID- 3050330 TI - [Bowen's disease of the palmar region. Apropos of a case]. AB - The authors study a case of Bowen's disease localized on the palm of the right hand, in a patient of the female sex, 63 years old. At the studied case the patient had presented squamous carcinoma in situ of the cervix which appeared about 7 years before the appearance of the cutaneous lesion. A literature revision was done, and the authors verified the rarity of the palm localization of the Bowen's disease. PMID- 3050329 TI - [Basocellular carcinoma in a smallpox vaccination scar]. AB - A 52 year old housewife was vaccinated against smallpox at the age of 18, on her right deltoid area. At the age of 50 she noticed erythema and scaling on the vaccination scar and 2 years later a nodule appear that enlarged during the following 3 months. There was no history nor skin changes suggestive of significant sun exposure. The histological examination of an initial biopsy and of the subsequently excised lesion revealed a basal cell carcinoma of the solid type. The relevant literature was reviewed and discussed with emphasis on sex and age incidence, age and site of vaccination, free interval between inoculation and tumor appearance, coexistence or not of other sun induced neoplasias and precancerous lesions and other possibly relevant clinical and etiopathogenetic aspects. PMID- 3050331 TI - [Smooth muscle hamartoma or nevus of Becker? Apropos of 4 cases]. AB - Theoretically, Becker's melanosis or hairy epidermal nevus and smooth muscle hamartoma are two quite separate entities. In fact, there are cases which could be considered as intermediate. As a matter of fact, a slight underlying smooth muscle hyperplasia can be seen in Becker's nevus; on the other hand, hypermelanosis of the basal layer and hypertrichosis may be encountered in smooth muscle hamartoma. The 4 here reported cases are examples of diagnostic difficulties for which the sometimes not clear-cut limits of these 2 types of lesion can be responsible. PMID- 3050333 TI - [Effects of UV-A on pig skin]. AB - The effects of high and progressive doses of UVA are studied on skin, from a clinical and microscopic, both optic and electronic, point of view. We use pigskin as pattern, due to its similarity to human skin and a PUVA 200 Waldman as source of radiation. PMID- 3050334 TI - [Localized Darier's disease. Topical treatment with retinoic acid]. AB - We report a new case of localized Darier's disease or keratosis follicularis with successful response to topical retinoic acid treatment. We comment the clinical histopathological features and differential diagnosis of these uncommon variant which has only been reported in 10% of the patients. PMID- 3050332 TI - [Epidermolysis bullosa and congenital skin aplasia (Bart's syndrome). Report of 3 cases]. AB - The association of epidermolysis bullosa (EB), congenital localized absence of skin and nail alterations like anonychia and dystrophy has been denominated Bart's syndrome, which was described nineteen years ago, and associated with simple, junctional and dystrophies epidermolysis bullosa. We explain in this study three cases, which because of their clinic characteristics will correspond to this new entity. All of these cases happened in the city of Trujillo, Peru. PMID- 3050335 TI - [Nail dystrophy of the big toe. Description of a clinical case]. AB - We report on four cases of great toenail dystrophy. This malformation presented itself in the first months of life. Involvement of other toenails and also of fingernails has been observed. In some of the cases spontaneous involution was referred. Pathogenesis, differential diagnosis and therapy are discussed. PMID- 3050337 TI - [Primary nodular amyloidosis of the tongue]. AB - Two cases of nodular primary amyloidosis of the tongue are reported. A review of the literature is made. PMID- 3050336 TI - [Evaluation of the efficacy and toxicity of local fusidic aid versus oral dicloxacillin in infections of the skin]. AB - In a double-blind test it was shown the efficacy of topical fusidic acid in comparison with oral dicloxacillin, in patients with skin infection by Staphylococcus aureus or/and Streptococcus pyogenes. In the experimental group, 16 patients from 20 received 2% topical fusidic acid in cream and reacted well in a shorter period of time than 17 from a group of 20 that had 500 mg. of dicloxacillin orally twice a day. PMID- 3050339 TI - [Insufficiency of the cervical lymph vessel system; lymphostatic encephalopathy and retinopathy following cervical block dissection]. AB - There are no lymphatic vessels in the CNS, in the retina and in the optic disk. In spite of this, however, cervical lymphostasis will result in lymphostatic encephalopathy and ophthalmopathy. PMID- 3050338 TI - [Entomophthoromycosis. Review]. AB - A review of the published reports on entomophthoromycosis (rhynoentomophthoromycosis and basidiobolomycosis) is presented. Geographic distribution, clinical, pathologic, mycologic and therapeutic aspects are emphasized. Primary and secondary visceral involvement are discussed. Data concerning 33 Latin American cases are analysed: 21 cases of rhynoentomophthoromycosis and 11 of basidiobolomycosis. Of these cases, 84% were described in Brazil and of the seven reported cases with primary visceral involvement reported in the world literature six were from Brazil. The importance of an early diagnosis of these infections is asserted specially in view of the resulting deformities and even death. PMID- 3050340 TI - [New aspects in the pathogenesis and therapy of hyperreflexive rhinopathy]. AB - Hyperreflectory rhinopathy (HR) is a non-specific hyperreactivity of the nasal mucosa. It causes hypersecretion, decreased nasal patency, sneezing and sometimes headache by local reflexes, which are beyond voluntary control. The synonymous name "vasomotor rhinitis" or "vasomotor rhinopathy" is no longer adequate with regard to our present state of knowledge of the autonomous innervation of the human nasal mucosa. Recent pharmacological investigations show the great importance of peptidergic neurons. In our own studies, the presence and the topical effects of the neuropeptide substance-P in human nasal mucosa are examined. A new concept of the autonomous innervation of the human nasal mucosa is presented. Apart from adrenergic and cholinergic neurons, it also includes peptidergic neurons (SP, CGRP, NKA, VIP, PHI, APP, GRP). According to this model, a hypothesis on the pathogenesis of HR is developed. A key position is occupied by the so-called "axon reflex" which is mediated by substance-P immunoreactive nerve fibers. It is released by a chemical, thermal or mechanical irritation. This axon reflex mediates pain, vasodilation and plasma extravasation (neurogenic oedema), hypersecretion such as smooth muscle contraction, and sneezing reflex. Capsaicin (8-methyl-N-vanillyl-6-nonenamid) leads to a selective degeneration of substance-P immunoreactive nerve fibres and desensitisation of its receptors after repeated topical or systemic application, thus blocking the axon reflex. The risk-free application of capsaicin was shown in self-experiments and in volunteers. Our hypothesis was confirmed by the good results of the treatment of a group of volunteer patients who suffered from HR.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3050341 TI - The anatomical basis for post-tracheotomy innominate artery rupture. AB - Classical teaching suggests that placement of a tracheostomy tube through the second or third tracheal rings will safeguard the innominate artery in the majority of patients. A tracheotomy was performed on ten fresh, adult cadavers through a vertical incision in the second and third tracheal rings. A series of measurements was made evaluating the relationship of the innominate artery to the tracheostomy tube and to laryngotracheal structures. In every dissection, some part of the tracheostomy tube cuff or tip was found to be adjacent to the innominate artery. We conclude that, contrary to general belief, placement of the tracheotomy incision at the second and third tracheal rings will not in and of itself protect the innominate artery from rupture. PMID- 3050343 TI - Laser instrumentation: a microtenaculum for laser tissue fusion. AB - Precise tissue apposition and edge eversion are essential for tissue fusion with the CO2 laser. A microtenaculum has been designed to accomplish this without the placement of horizontal mattress stay sutures. This instrument permits tissue orientation and approximation for primary laser fusion as well as for the repair of small gaps present after initial welding. PMID- 3050342 TI - Vasovasostomy in the murine vas deferens: comparison of the Nd:YAG laser at 1.06 microns and 1.318 microns to the CO2 laser. AB - A comparison is made of laser anastomoses of the murine vas deferens at different energies with the neodymium (Nd):YAG laser at 1.06 micron and 1.318 micron and with the CO2 laser. A total of 28 welds were performed with a free-hand technique employing a 600-micron silicon fiber with the Nd:YAG and a hand piece with a 500 micron spot size for the CO2. After 6 weeks, all animals were sacrificed and the vasa evaluated for patency. Fifteen out of 28 controls repaired with microsurgical techniques were found to be patent; 4/10 vasa were patent with use of the Nd:YAG at 1.318 micron at laser energies of 300 mW and 500 mW. At 1.06 micron, only 1/4 anastomoses was patent at a power setting of 1 W. None of the anastomoses performed with the CO2 laser was patent. Histologic study revealed intense fibrosis in all the lasered vasa, with sperm granuloma formation associated with most anastomoses. Although this is a preliminary study, it appears that the Nd:YAG laser at 1.318 micron and a power setting of 300-500 mW provides patency rates superior to the Nd:YAG at 1.06 micron and to the CO2 lasers and is equivalent to standard micro-surgical techniques in the murine vas deferens. PMID- 3050344 TI - Characterization of the radioiodinated analogue of SCH 23390: in vitro and in vivo D-1 dopamine receptor binding studies. AB - A new radioiodinated molecule, 125I-SCH 38840 (previously referred to as 125I-SCH 23982), has been recently reported to be a D-1 dopamine receptor ligand. The current study confirms and expands the characterization of both the radiolabeled and unlabeled forms of this compound, as well as describing the development of an in vivo D-1 receptor binding assay utilizing the 125I-SCH 38840. The binding of 125I-SCH 38840 to rat striatal membranes, in vitro, was saturable and exhibited a KD of 1.47 nM. Competition studies using 125I-SCH 38840 exhibited a pharmacological profile consistent with the proposal that 125I-SCH 38840 was binding to the D-1 receptor. Further studies with the unlabeled SCH 38840 demonstrated that it inhibited dopamine-stimulated adenylate cyclase with a KI of 66.1 nM, indicating that SCH 38840 was acting as a D-1 antagonist. Behavioral studies demonstrated that SCH 38840 (MED = 1.0 mg/kg, s.c.) blocked conditioned avoidance responding in rats, a measurement considered predictive of anti psychotic activity in man. In vivo binding of 125I-SCH 38840 to rat striatum following s.c. administration was specific. Peak striatal levels were observed 1 h after injection, with measurable binding observed out to 8 h post-treatment. The displacement of the in vivo binding by unlabeled standards again suggested a D-1 selective interaction. The half-life of the in vivo binding of 125I-SCH 38840 was approximately 1.25 h, and was nearly equivalent to the half-life of the anti CAR activity of unlabeled SCH 38840. These results clearly demonstrate the D-1 nature of SCH 38840's behavioral activity and strengthen the anti-psychotic potential of a D-1 antagonist. PMID- 3050345 TI - Geraniol interferes with membrane functions in strains of Candida and Saccharomyces. AB - Geraniol, an olefinic terpene, was found to inhibit growth of Candida albicans and Saccharomyces cerevisiae strains. Geraniol was shown to enhance the rate of potassium leakage out of whole cells and also was shown by fluorescence polarization to increase C. albicans membrane fluidity. Biophysical studies using differential scanning calorimetry, fluorescence polarization and osmotic swelling of phospholipid vesicles demonstrated that geraniol decreased the phase transition temperature of dipalmitoylphosphatidylcholine vesicles, affected fluidity throughout the bilayer, particularly the central portion of the bilayers, and caused an increase in bilayer permeability to erythritol. Geraniol may have potential use as an antifungal agent. PMID- 3050347 TI - The 1988-89 leader for Maryland medicine: Michael R. Dobridge MD. PMID- 3050349 TI - A directory of medical software companies. PMID- 3050348 TI - HIV in women and their pregnancies. PMID- 3050350 TI - Retracted confessions: legal, psychological and psychiatric aspects. PMID- 3050346 TI - Sunlight, melanogenesis and radicals in the skin. AB - Melanocytes are cells of neural crest origin residing at the dermal-epidermal juncture. They produce specialized organelles called melanosomes within which the biochemical processes of melanization occurs. UV radiation is capable of inducing melanogenesis and, during the biosynthesis of melanins, several of the putative precursors "leak out" of the melanosome and can be detected in the skin, serum and urine of individuals undergoing active melanogenesis. Most notable are the cysteinyldopas (formed by nucleophilic addition of cysteine to dopaquinone) and several dihydroxyindoles (formed by intramolecular cyclization of dopaquinone). These catechols often are methylated in the melanocyte to afford a mixture of the monomethoxy derivatives and, in some cases, the dimethoxy species. Recent investigations in our laboratories have demonstrated that the cysteinyldopas, dihydroxyindoles, and their various methylated derivatives are photochemically unstable. Irradiation with biologically relevant ultraviolet radiation (i.e. wavelengths greater than 300 nm) results in the rapid destruction of the precursors/metabolites and the production of a variety of free radical species. The photochemistry and potential photobiological significance of melanogenic intermediates is discussed. PMID- 3050351 TI - Resuscitation artefact. PMID- 3050352 TI - Implications of exercise testing for prediction of athletic performance: a contemporary perspective. AB - One of the most fundamental beliefs in exercise physiology is that performance during maximum exercise of short duration is limited by the inability of the heart and lungs to provide oxygen at a rate sufficiently fast to fuel energy production by the active muscle mass. This belief originates from work undertaken in the 1920's by Hill and Lupton. A result is that most, if not all, of the studies explaining the effects of exercise training or detraining or other interventions on human physiology explain these changes in terms either of central adaptations increasing oxygen delivery to muscle or of peripheral adaptations that modify the rates of oxygen or fuel utilization by the active muscles. Yet a critical review of Hill and Lupton's results shows that they inferred but certainly did not prove that oxygen limitation develops during maximal exercise. Furthermore, more modern studies suggest that, if such an oxygen limitation does indeed occur during maximal exercise, it develops in about 50% of test subjects. Thus, an alternative mechanism may need to be evoked to explain exhaustion during maximal exercise in a rather large group of subjects. This review proposes that the factors limiting maximal exercise performance might be better explained in terms of a failure of muscle contractility ("muscle power"), which may be independent of tissue oxygen deficiency. The implications for exercise testing and the prediction of athletic performance are discussed. PMID- 3050353 TI - Characterization of muscles injured by forced lengthening. I. Cellular infiltrates. AB - Myofiber injury-repair was studied in rat soleus muscles to elucidate the role of infiltrating cells in the injury-repair process. Muscle injury was induced by forced muscle lengthening with the contralateral muscle serving as a control. The muscles were removed for histologic, histochemical and immunohistochemical procedures at varying periods (12-120 h) post-injury. All injured muscles were severely damaged with many cells present in the interstitial spaces between myofibers. Normal appearing myofibers demonstrated elevated lysosomal proteolytic activity, but no evidence of increased activity, indicative of phagocytic cells, was found in or between damaged myofibers. The esterase stain for macrophages and immunohistochemical techniques for mast cells also provided no support for either cell type predominating in the damaged area, although mast cell degranulation could be observed in the pericapillary regions. In contrast, the use of a specific antisera for a multicatalytic protease uniquely defined most of these cells as myogenic in origin. They appeared to be most numerous between the torn ends of a myofiber. Surprisingly, the remainder of the cells appeared to be of lymphoid origin. PMID- 3050354 TI - Characterization of muscles injured by forced lengthening. II. Proteoglycans. AB - After forced muscle lengthening of rat soleus muscle, alterations in muscle connective tissues were monitored by fluorescent immunohistochemical methods. Monoclonal antibodies directed against the polysaccharide attachment region of proteoglycans were used to observe changes in localization of 4-sulfated, 6 sulfated, or unsulfated chondroitin sulfate disaccharide units covalently bound to the proteoglycan protein core after injury. Additionally, fluorescein-labeled concanavalin A lectin and polyclonal antiserum to heparan sulfate proteoglycan were also localized in muscle sections during the regenerative process over 5 days after injury. Although proteoglycan localization was absent at or near the site of myofiber damage after injury, some distinct basal lamina remained as a matrix for regenerating myofibers. By the fifth day post-injury, the localization of these matrix components had returned to that seen in uninjured soleus muscles. The physiological significance of these extracellular matrix changes appeared to center on the repair of the torn myofiber and indicate an interdependence between myofibers and the extracellular matrix in this type of regeneration. PMID- 3050356 TI - Quantitative flow measurements on phantoms and on blood vessels with MR. AB - A broad spectrum of MR methods has been published in the last few years for the visualization and the quantification of flow in human blood vessels. We describe the results of an extended study to measure the flow quantitatively using the MR phase modulation method. A gradient echo sequence which allows short repetition times was synchronized with the ECG. The instantaneous velocity profiles and the integrated flow rate can be determined throughout the cardiac cycle in up to 45 consecutive time intervals. The instantaneous velocity distributions can be displayed as 2D profiles over the lumen of the blood vessel. The technique has been successfully evaluated with a set of phantom experiments. Following this, quantifications of the blood flow rates in vivo were performed on the abdominal aortas of healthy volunteers. The MR flow results were compared to those obtained with conventional Doppler ultrasound examinations. The results of both methods are in good quantitative agreement. PMID- 3050355 TI - A method for the assessment of area-elastic surfaces. AB - A method is presented for the evaluation of area-elastic surfaces using two subject tests. Deformations and jump heights are determined for 12 sprung floor samples to accuracies of 0.12 mm and 0.4 cm respectively, using filming techniques. It is found that the deformation of a floor sample is dependent on both the subject group and the dimensions of the floor sample. The method showed significant differences in deformation values of the 12 floor samples but only small differences in jump heights. PMID- 3050357 TI - Recent microsurgical advances in China. AB - For more than two decades, microsurgery has been developed, practiced, and refined in China. For the most part, this has taken place independent of a parallel process in the Western World. Only until recent years has there been some communication from China regarding this microsurgical experience. The authors believe that there is much yet to be learned from the wealth of Chinese experience in microsurgery, just as Chinese surgeons have gained valuable information from the West. The authors reviewed the microsurgical literature in China from 1983 to 1986, and present here what we regard to be of particular interest to American microsurgeons, including the areas of digital reconstruction, replantation, flap transfer and transposition, and microsurgical techniques and patient management. Some historical background in the development of Chinese microsurgery is also provided. PMID- 3050358 TI - Microsurgery in China and the development of Western reconstructive microsurgery. AB - The development of extremity replantation and reconstructive microsurgery was initially carried out independently among the pioneering centers throughout the world. The comparative development of these techniques is outlined. With the fast growth of microsurgical procedures, many of the U.S. residency programs have incorporated microsurgery training into the curriculum. PMID- 3050359 TI - Extracellular proteins of Vibrio cholerae: nucleotide sequence of the structural gene (hlyA) for the haemolysin of the haemolytic El Tor strain 017 and characterization of the hlyA mutation in the non-haemolytic classical strain 569B. AB - The EI T or haemolysin, product of hlyA, is exported from Vibrio cholerae as a Mr 80,000 protein which can be subsequently cleaved to give two proteins of Mr 65,000 and 15,000. Nucleotide sequence analysis has demonstrated that hlyA encodes a protein of Mr 82,250 with a potential 18-amino-acid signal sequence. The non-haemolytic classical strain 569B has been shown to have a structural gene defect rather than a defect in secretion. By non-reciprocal recombination it was possible to transfer this defect onto a plasmid and show that a truncated hlyA product of Mr 27,000 is made in Escherichia coli K-12 minicells. Nucleotide sequence analysis demonstrates an 11-base-pair deletion which would result in a Mr 26,940 protein probably loosely associated with the membrane. PMID- 3050360 TI - Overexpression, solubilization and refolding of a genetically engineered derivative of the penicillin-binding protein 3 of Escherichia coli K12. AB - Replacement of the amino-terminal 40-amino-acid region of the 588-amino-acid precursor of the membrane-bound penicillin-binding protein 3 (PBP3) by the decapeptide MKGKEFQAWI was carried out by altering the amino-coding end of the ftsI gene. Insertion of the modified gene into a runaway-replication plasmid under the control of a fused lpp promoter and lac promoter/operator, resulted in the overexpression by Escherichia coli of the modified PBP3 (designated PBP3**) in the cytoplasm. About 80% of the accumulated PBP3** underwent sequestration in the form of insoluble protein granules that were isolated by cell breakage or cell lysis. After selective removal of contaminants by an EDTA-lysozyme/DNase (deoxyribonuclease)/Nonidet extraction, treatment of the granules with guanidinium chloride followed by dialysis against buffer containing 0.5 M NaCl yielded a refolded, water-soluble PBP3**, which, upon chromatography on Superose 12, exhibited the expected 60,000 molecular mass. The refolded PBP3** bound benzylpenicillin in a 1 to 1 molar ratio, was highly sensitive to aztreonam and showed the same degree of thermostability, in terms of penicillin-binding capacity, as the parent, membrane-bound PBP3, suggesting that protein refolding occurred with formation of the correct intramolecular interactions. Two to three mg of refolded PBP3** can be obtained from 1 litre of culture of the overproducing strain. PMID- 3050361 TI - ICF/general: an alternative for older ICF/MR residents with geriatric care needs. PMID- 3050362 TI - Self-injurious behavior, its treatment, and normalization. PMID- 3050364 TI - Effects of an oral antidiabetic drug on the fibrinolytic system of blood in insulin-treated diabetic patients. AB - Selected variables of the fibrinolytic system were assessed in 23 men with insulin-treated diabetes with no measurable pancreatic beta-cell function. Gliclazide, a second-generation sulphonylurea drug, was administered to the patients over a period of 6 months in daily doses of 160 mg or 240 mg, and blood samples were obtained before, during, and after treatment. Determined by global assays, the drug did not significantly change plasminogen activator activities in euglobulins. Measurements of specific components of the system of fibrinolysis showed a marginal increase during administration of gliclazide of tissue-type plasminogen-activator antigen and prekallikrein activity in plasma, whereas the activities in euglobulins of the intrinsic plasminogen proactivators remained nearly the same during the study. Levels in plasma and euglobulin of C1 inactivator antigen and in plasma of factor XII antigen and t-PA inhibition capacity remained constant throughout the study. There were no changes of the increase in concentration of t-PA activity and t-PA antigen following venous occlusion. The metabolic state remained the same during the whole study. It is concluded that gliclazide induces small, but significant, non-insulin-dependent extrametabolic effects on the extrinsic (t-PA) and intrinsic (prekallikrein) system of fibrinolysis. Whether these changes are of physiological importance remains to be demonstrated. PMID- 3050363 TI - The glucose stimulus-response curve of the beta-cell in physically trained humans, assessed by hyperglycemic clamps. AB - In order to examine the effect of habitual exercise on beta-cell responses over a wide range of plasma glucose levels, plasma insulin and C-peptide responses to 2 1/2-hour hyperglycemic clamps at 7.5, 10, and 15 mmol/L glucose were assessed in six trained athletes and six age- and weight-matched sedentary controls. Athletes were significantly fitter than controls (estimated maximal oxygen uptake [VO2 max] mean 44 v 30 mL.kg-1.min-1, P less than .05) and were more sensitive to insulin as assessed by dividing the mean glucose infusion rate over the last 20 minutes of the clamp by the steady-state plasma insulin (mean 0.44 v 0.19 mg.min 1.kg-1.nmol-1. L, respectively, P less than .01). Plasma C-peptide responses were lower in the athletes, both fasting (geometric mean 0.28 v 0.62 nmol/L, P less than .05), and at the end of all clamps (at 7.5, 10, and 15 mmol/L plasma glucose, respectively, 0.65 v 1.43, 1.25 v 2.85, and 2.40 v 4.46 nmol/L each, P less than .05). First-phase plasma C-peptide responses were lower in the athletes at the 10 and 15 mmol clamp levels. The slope of the glucose-C-peptide stimulus response curve was approximately linear over the range examined, the slope being significantly shallower in athletes than controls for both first phase (P less than .01) and second phase (P less than .01). Plasma insulin responses were similar to C-peptide responses. The attenuation of beta-cell responsiveness over a wide glucose range may be an adaptation to the enhanced peripheral insulin sensitivity seen in athletes. PMID- 3050365 TI - Lipodystrophic diabetes mellitus. Investigations of lipoprotein metabolism and the effects of omega-3 fatty acid administration in two patients. AB - We investigated the metabolic effects of omega-6 (safflower oil) and omega-3 (fish oil) fatty acid-enriched diets (65% carbohydrate, 20% fat) in two patients with a syndrome of diabetes mellitus, lipodystrophy, acanthosis nigricans, chylomicronemia, and abdominal pain. 3H-glycerol was used to evaluate triglyceride-rich lipoprotein-triglyceride (TRLP-TG) metabolism, and changes in glucose and insulin dynamics were also studied. On the omega-6 diet, both subjects demonstrated four- to five-times normal rates of TRLP-TG production and glycerol biosynthesis, and striking decrements in the fractional catabolic rate (FCR) for TRLP-TG and TRLP-particles. Both subjects had elevations in nonesterified fatty acid (NEFA) concentrations. In one patient, the omega-3 diet markedly decreased serum triglycerides and newly synthesized triglyceride glycerol production, in association with a fall in NEFA. In both subjects, plasma glycerol reutilization for triglyceride synthesis, normal on the omega-6 diet, was abolished on the omega-3 regimen. Plasma postheparin lipolytic activity was normal on both diets. On the omega-3 diet, xanthomas and hepatomegaly decreased and, in the patient who had no reduction in serum triglycerides, pancreatitis attacks virtually ceased. Mean 24-hour serum glucose levels were higher, and both basal and peak C-peptide responses to a carbohydrate meal were blunted on the omega-3 diet. One patient became ketonuric. We conclude the cause of hypertriglyceridemia in these patients was due to increased lipid synthesis and hypothesize that this is secondary to high plasma concentrations of NEFA. In addition, an omega-3 diet in these subjects inhibited insulin secretion and worsened glucose tolerance. PMID- 3050367 TI - Impaired insulin action in adipocytes of New Zealand obese mice: a role for postbinding defects in pyruvate dehydrogenase and insulin mediator activity. AB - This study investigated possible mechanisms underlying insulin resistance in the New Zealand Obese (NZO) mouse, an animal model for obese, non-insulin-dependent diabetes. Insulin binding, mediator generation, and action both at the level of glucose utilization and enzyme modulation were compared in adipocytes from lean control New Zealand Chocolate (NZC) mice and NZO mice during the development of the syndrome. Abnormalities of insulin stimulation of glucose transport and utilization in NZO mouse adipocytes were found which involved both decreased sensitivity and responsiveness to insulin. The defects were evident at an early age (4 weeks) and could not be attributed to differences in nonstimulated glucose metabolism, which was similar in the control NZC and obese NZO strains of mouse. Insulin binding to its receptor was only moderately decreased in adipocytes of NZO mice. Pyruvate dehydrogenase (PDH) activity of NZO mouse adipocytes was totally unresponsive to insulin in contrast to the impaired but still significant insulin stimulation of glucose transport and utilization, suggesting a postreceptor defect at the level of insulin stimulation of this enzyme. Insulin stimulated the production of a low molecular weight factor which activated pyruvate dehydrogenase in NZC mouse adipocytes (insulin mediator) but, paradoxically, caused a decrease in mediator production or activity in adipocytes from NZO mice. Thus, insulin either inhibited mediator production or stimulated generation of an inhibitory mediator in adipocytes from this strain. No evidence for the latter mechanism was found. This study demonstrates in adipocytes of NZO mice: (1) a receptor defect and (2) a postreceptor defect of insulin action at the level of pyruvate dehydrogenase activation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3050366 TI - Pancreatic hypersensitivity to glucose by young obese Zucker rats (fa/fa). AB - Insulin secretory response to glucose was investigated in 5- to 6-week-old male Zucker obese (fa/fa) and lean (Fa/Fa) rats using a pancreatic perfusion procedure. Blood glucose response to fasting was studied in lean and obese animals over 24 hours. Plasma glucose was slightly elevated in pentobarbital anesthetized obese rats. However, plasma insulin was 4.6 times greater than that of leans. A hypoglycemic glucose stimulus (75 mg/dL) caused pancreata from obese animals to release 6 times more insulin than lean animals. Stimuli of 125 mg/dL (normoglycemic) and 600 mg/dL (hyperglycemic) caused hypersecretion of 8 and 5 times, respectively. Hypersecretion was not accounted for solely by the twofold increase in pancreatic insulin content. Obese animals had steeper decreases in plasma glucose than lean controls during seven to 13 hours of fasting. Hypersecretion by pancreata from young obese rats to physiological levels of glucose may result in hyperphagia in order to maintain normoglycemia. PMID- 3050368 TI - Regulation of hepatic glucose output during moderate exercise in non-insulin dependent diabetes. AB - In normal subjects during moderate exercise there is a strong negative correlation between plasma glucose and hepatic glucose output (HGO) suggesting a negative feedback regulation of HGO by plasma glucose. Little information is available about HGO responses to exercise in non-insulin-dependent diabetes mellitus (NIDDM). To determine whether the same feedback relationship is operative, we have compared the glucose turnover responses to moderate exercise (50% Vo2max for 60 minutes) of nonobese non-insulin-dependent diabetic subjects (NIDDM, n = 7) with a group of age-matched controls (n = 5). Glucose turnover responses to exercise in NIDDM were heterogeneous. Plasma glucose showed sustained falls, no change, or sustained rises in different individuals. Similarly, HGO responses ranged from undetectable to responses comparable to those of normal subjects. The mean integrated HGO response in NIDDM was significantly reduced compared with controls (11 +/- 6 [SEM] v 33 +/- 7 mmol/h/70 kg, P less than .05); mean glucose utilization response was also reduced but not significantly different from controls (NIDDM 18 +/- 5 v control 35 +/- 6). In NIDDM there was no significant feedback-control relationship between plasma glucose and HGO (r = -0.20, P = NS) in contrast to controls (r = -0.87, P less than .01). We conclude that feedback control of HGO by plasma glucose during moderate exercise is impaired in NIDDM. This impairment may be due to defective nonpancreatic glucoregulatory mechanisms. PMID- 3050369 TI - Insulin inhibits protein degradation in skeletal muscles of eviscerated fasted rats. AB - The inhibitory effect of insulin on skeletal muscle protein breakdown was tested in fed or fasted rats whose muscles had been isolated by evisceration. Degradation was estimated from the accumulation of tyrosine in the plasma after protein synthesis was blocked with cycloheximide. According to earlier results from this method, fasting for 20 hours increases proteolysis. In the present work, in fasted preparations whose plasma insulin concentrations were maintained at higher levels by intravenous infusions of insulin and glucose, proteolysis was inhibited by insulin. In fed eviscerated rats, insulin was ineffective. To examine more closely the response of fasted skeletal muscles to insulin, plasma insulin concentrations in preparations of 20-hour fasted rats were varied over a wide range by treatment with an initial bolus of insulin, and then proteolysis was measured as change in tyrosine concentration during a 90-minute period. In fasted rats, insulin inhibited proteolysis in a dose-dependent fashion, with a half-maximum concentration of 35 microU/mL. However, at insulin levels equal to concentrations found in normal fed rats, compensation for the fast-induced proteolytic effect was still incomplete. The results suggest that in these more intact preparations, insulin does have a physiologically significant inhibitory effect on fast-induced muscle protein degradation, but that changes in fasting and feeding cannot be entirely explained by the action of insulin. PMID- 3050371 TI - Cumulative subject index. Volumes 102-119, 121-134. PMID- 3050372 TI - Pro. M.D.--a diagnostic expert system shell for clinical chemistry test result interpretation. PMID- 3050370 TI - Relationship between lipoprotein levels and in vivo insulin action in normal young white men. AB - In epidemiologic studies, hyperinsulinemia has been found to be an independent risk factor for coronary heart disease (CHD). However, the mechanisms responsible for its role in atherogenesis remain unclear. We studied the relationship of in vivo insulin action and plasma lipids and lipoproteins in 44 normotriglyceridemic white men (aged 18 to 34 years). The euglycemic, hyperinsulinemic glucose clamp technique was used to quantitate insulin-mediated glucose disposal (M/I value) at a plasma insulin concentration of approximately 100 microU/mL. The M/I value correlated negatively with plasma triglycerides (r = -0.553, P less than .0001), as well as with fasting plasma insulin levels (r = -0.483, P less than .001), independent of age, body mass index, and fasting plasma glucose levels. A negative correlation of the M/I value was also observed with very low density lipoprotein (VLDL)-cholesterol (r = -0.347, P less than .05), VLDL-triglycerides (r = -0.474, P less than 0.005), and total cholesterol/high density lipoprotein (HDL)-cholesterol ratio (r = -0.431, P less than .01). The relationship between the M/I value and the total cholesterol/HDL-cholesterol ratio was independent of VLDL-cholesterol and VLDL-triglycerides, however, not independent of plasma triglycerides. No relationship was observed between insulin-mediated glucose uptake and total cholesterol, low density lipoprotein (LDL)-cholesterol, and HDL cholesterol values. Individual differences in plasma triglycerides, fasting insulin concentration, and the total cholesterol/HDL-cholesterol ratio accounted for about half the variance observed in the M/I value.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3050373 TI - Multiple typing of Salmonella typhimurium isolates: an epidemiological study. AB - An epidemiological study was carried out on sixty-four Salmonella typhimurium strains isolated in Palermo during the period January-July 1987 and identified at the Southern Italy Center of Enterobacteriaceae. These included 5 isolates from a small food-poisoning outbreak, which resulted antibiotic susceptible, not colicinogenic and untypable by the phage-type scheme of Anderson. Plasmid profile analysis was not a reliable method to differentiate them from non epidemic strains. The 5 epidemic isolates, belonging to biotype 25a, were assigned into NT 2 phage-type by an accessory set of phages developed in this laboratory. Such biotype/phage-type association was never detected in the remaining Salmonella typhimurium strains isolated during the first 7 months of 1987. Chromosomal DNA analysis provided additional information on the relationships among Salmonella typhimurium isolates. PMID- 3050374 TI - Production of Linnocuicina 819, a bacteriocin produced by Listeria innocua. AB - We studied the reciprocal antagonistic properties of 14 Listeria strains. Listeria innocua 29-CCM-A 819 (CIP 8011) produces a bacteriocin named Linnocuicina 819. This substance is mainly released in the exponential phase of growth and was isolated from the supernatant of TPB cultures. Thermal resistance and sensitivity to proteolytic enzymes was assayed. Linnocuicina 819 has a bactericidal mode of action producing a lytic effect. PMID- 3050375 TI - Scanning electron microscopy of Serratia marcescens producing prodigiosin. AB - Production of high concentrations of prodigiosin by growing cells of Serratia marcescens was accompanied by the formation of extracellular protrusions as was revealed by scanning electron microscopy. Prodigiosin extracted from the bacterium was compared with the extracellular material. Bacteria which did not produce prodigiosin showed no extracellular protrusions. PMID- 3050376 TI - Formation of a hexagonal lattice structure by an R-form lipopolysaccharide of Klebsiella: effect of various divalent cations on the lattice formation. AB - The R-form lipopolysaccharide from Klebsiella pneumoniae strain LEN-111 (O3-:K1 ), from which cationic material had been removed by electrodialysis, was previously shown to form a hexagonal lattice structure with the lattice constant of 14 to 15 nm when suspended in 50 mM tris(hydroxymethyl)aminomethane buffer at pH 8.5 containing 10 mM Mg2+. Under this experimental condition, effects of other divalent metal cations on the hexagonal assembly of the electrodialyzed LPS were compared with that of Mg2+. The Zn2+, Hg2+, Cu2+, and Ni2+ could produce essentially the same hexagonal lattice structure with the lattice constant of 14.5 to 15.0 nm as that formed with Mg2+. The Cd2+, Co2+, and Fe2+ produced the hexagonal lattice structure with the lattice constant of 15.5 to 16.0 nm, and Ba2+, Sr2+, and Ca2+ produced that with the lattice constant of 18 to 19 nm. In addition, the hexagonal lattice structures formed with the latter three cations were less orderly than those formed with the other cations. When the higher concentrations of Ba2+, Sr2+, and Ca2+ were used, the lattice constants were not shortened. The length of lattice constants of the hexagonal lattice structures formed with the divalent cations did not relate to the quantity of the cations bound to the LPS. Among the divalent cations tested, Hg2+ was bound to the LPS in the smallest amount (its atomic ratio to P, 0.07), and Zn2+ and Fe2+ were bound in very large amounts (their atomic ratios to P, 2.94 and 8.28, respectively). PMID- 3050377 TI - Replication of human immunodeficiency virus in two T-cell lines and their application as antigens for immunofluorescence. AB - Two T-cell lines, TALL-1 and CCRF-CEM, were infected with human immunodeficiency virus (HIV), strain LAV, to explore the time course of the appearance of various virus specific antigens, and to establish an antibody assay system by indirect immunofluorescence (IF). These cells were infected with LAV at two different input multiplicity of infection (MOI). Antigens were tested by Western blot analysis (WB) and IF. Antigens for WB were extracted from the infected cells at various times after infection, but pooled sera of American HIV carriers could not recognize gp41 or gp160. Antigen expression was highest in CCRF-CEM, but, as the antigen for IF, TALL-1 infected at the MOI of 8.0 was the most suitable 7 days after infection, because it includes a fairly large number of uninfected cells, which served as the internal control. PMID- 3050378 TI - A case of lung infection caused by an unusual strain of Nocardia farcinica. AB - A case of lung infection caused by an unusual strain of Nocardia farcinica is reported. This is the third case of the N. farcinica infection in this country. The strain failed to utilize rhamnose as sole carbon source, but could be identified by a numerical identification method. The mycolic acids contained 1-3 double bonds and the numbers of the carbon atoms of the mycolic acids were 50 to 60, average 56. PMID- 3050379 TI - Production and partial purification of a fluid-accumulating factor of non-O1 Vibrio cholerae. AB - A fluid-accumulating factor (FAF in the ligated rabbit ileal loop test) from a strain of non-O1 Vibrio cholerae not producing cholera toxin-like enterotoxin (CTLT) was partially purified by ammonium sulfate precipitation, gel filtration with Sephadex G-100, and DEAE cellulose column chromatography. The preparation thus obtained showed collagenolytic, cytolytic, necrotic, and hemorrhagic activities, but was not lethal to mice nor hemolytic to sheep erythrocytes. Desquamation of epithelial cells, inflammatory edema, and hemorrhage were observed in sections of rabbit intestine after inoculation of partially purified FAF (PPFAF). Biological and enzymatic activities of FAF were completely neutralized with anti-PPFAF rabbit serum. More than 70% of non-O1 V. cholerae strains from human diarrheal feces produced FAF in the shake culture of heart infusion broth (Difco). A fluid-accumulating factor immunologically similar to FAF of non-O1 V. cholerae was also produced by V. mimicus strains isolated from human diarrheal feces. These results indicate that the FAF produced by CTLT negative non-O1 V. cholerae strains is an entity closely related to a cytolytic and hemorrhagic substance or the like, and that this FAF may play a role in the enteropathogenicity of CTLT-negative strains. PMID- 3050380 TI - Distribution of Clostridium botulinum in Japan and in Shinkiang district of China. AB - Soil specimens obtained from several areas of Japan, which are closely located to or facing the Continental land of China, were examined for the distribution of Clostridium botulinum, especially pertaining to types A and B. A total of 266 specimens of Japan, when cultured, showed no type A or B toxicity, although 30 (11.3%), 4 (1.5%), and 10 (3.8%) of the specimens showed C1, C2, and type E toxicities, respectively. On the contrary, types A and/or B toxicities were shown, by the same method, in 14 of 20 specimens of Shinkiang district, China. The highest number of C. botulinum cells found in one gram of soil specimen was 25 for type A and 10 for type B. PMID- 3050381 TI - Linkage of K99 production and STa activity in a plasmid of an Escherichia coli porcine isolate. AB - An Escherichia coli strain ZP118 of porcine origin was shown to harbor a 68 megadalton (Md) plasmid coding for a colonization factor K99 and heat-stable enterotoxin (STa), and a self-transmissible 51-Md plasmid coding for drug resistance. One of the transconjugants obtained by mating between ZP118 and E. coli C was found to harbor a 90-Md plasmid coding for K99, STa and drug resistance. Restriction endonuclease analysis suggested that the 90-Md plasmid could be a recombinant between the 68-Md plasmid and the 51-Md plasmid. PMID- 3050382 TI - Problem-solving skills, solving problems and problem-based learning. AB - This paper reviews the empirical evidence in support of the three concepts in the title. To the extent that a skill should be a general strategy, applicable in a variety of situations, and independent of the specific knowledge of the situation, there is little evidence that problem-solving skills, as described and measured in medical education, possess these characteristics. Instead there is an accumulation of evidence that expert problem-solving in medicine is dependent on (I) a wealth of prior specific experiences which can be used in routine solution of problems by pattern recognition processes, and (2) elaborated conceptual knowledge applicable to the occasional problematic situation. The use of problem based learning (PBL) as an educational strategy is explored. In particular, the evidence suggesting the compatibility of PBL with this theory of expertise is discussed. Finally, I review some issues in the design of PBL curricula from the perspective of the proposed model of expertise. PMID- 3050383 TI - Immunotherapy in acute myelogeneous leukemia (AML)--a tool for maintaining remission? AB - Twelve trials of adjuvant immunotherapy in patients suffering from AML have been analyzed and compared to results from experimental studies. The analysis presents evidence suggesting that the immune response to leukemia cells exists in a state of balance; this appears to be regulated by the dose of antigen and the state of the cells used to immunize the patients. The injection of high doses of live allogeneic leukemia cells produced a significantly prolonged duration of the first remission in in AML patients. Immunization with high doses of irradiated and dead cells induced some prolongation of the remission phase and survival time, although the percentage of survivors after 3 years was not increased in these groups as compared to non-immunized patients. Immunization by the same route using a 100-fold lower amount of leukemia cells afforded no protection against relapse of the disease during the maintenance phase. A few patients even developed the relapse earlier than did patients treated with chemotherapy alone. Our understanding of the immune responses to malignant cells has increased considerably during the past 2 decades due to various observations. The results obtained with active immunotherapy of the AML patients during the same period agree well with experience from laboratory studies. Thus the results confirm the potential of an immunological interaction between the leukemia cells and the patient. Consequently, it seems likely that the addition of immunotherapy to the treatment of patients with AML might be effective and the tool for maintaining the phase of remission by up-to-date immunological engineering. PMID- 3050384 TI - Chronic lymphocytic leukemic patients, resistant to chemotherapy. AB - In four reported studies, intensive chemotherapy of various types was administered to large groups of patients with chronic lymphocytic leukemia (CLL) in advanced stages. About half of the patients were non-responders to the therapy. Resistance to therapy was also found in 12% of patients with early CLL and in 19 CLL patients who had in vitro radioresistant lymphocytes before therapy. The findings support the hypothesis of the heterogeneity of CLL patients and suggest the need for new methods for therapy of non-responders and for in vitro or in vivo tests to diagnose non-responders. PMID- 3050385 TI - The potential significance of sulphated glycosaminoglycans as a common constituent of all amyloids: or, perhaps amyloid is not a misnomer. AB - Amyloid is a generic term referring to a group of diseases in which proteinaceous tissue deposits all have in common specific stain affinities, a common appearance in polarized light, common ultrastructure fibrillary characteristics, and uniform x-ray diffraction and infrared spectral properties. Where groups of diseases have a common underlying pathogenetic process the polypeptide responsible for the protein fibril is the same regardless of the specific disease. Where diseases have a different underlying pathogenesis the polypeptide is unique for each disease. The different amyloidogenic polypeptides are clearly not related in terms of amino acid sequence or function, yet they all tend to fold in such a way as to present the same staining, structural or spectral properties. It is proposed that amyloid fibrils are not only composed of the specific amyloidogenic polypeptide but also highly sulphated glycosaminoglycans or proteoglycans which have a profound influence on the manner in which the peptides fold and interact with each other. It is this highly charged carbohydrate which may be common to all amyloids and which plays a determining role in the final appearance of the deposit. Amyloid should therefore be considered as more than simply a protein entity but, as its name originally implied, one related to carbohydrate deposition as well. PMID- 3050386 TI - Carcinogenesis as the result of the deficiency of some essential trace elements. AB - "Energetic" biological trace elements [gallium (III), germanium (IV), silicon (silica), arsenic (V) and selenium (IV)] occurring in DNA of eukaryotic cells may improve the semiconductor properties of DNA and may influence the mechanisms that control genetic expression at the electronic level. Their roles are postulated as follows: (i) to maintain the level and direction of free sliding electrons in DNA, (ii) to modulate the electron conductivity and hole conductivity of DNA. This specific electronic nature of DNA take the form of magnetic pigeonholes in which an electric pulse is (0), or is not (1) stored as an area of local magnetisation. These types of conductivity occurring in different parts of DNA of different cells could participate in the switch on and switch off of genetic information in gene expression. This model may help to elucidate the mechanism of action of these naturally occurring antitumor agents and may help in understanding the role of trace elements in charge transport of DNA and in carcinogenesis. PMID- 3050387 TI - Zinc salts that may be effective against the AIDS virus HIV. AB - Zinc salts that can be safely given to humans in dosages that may be effective against the AIDS virus are reviewed. PMID- 3050389 TI - The artificial urinary sphincter: review and progress. AB - Urinary incontinence, the inability to retain urine, creates a misery that cannot be overestimated. The foul odor emanating from the patient repels family and friends to such an extent that it affects the social life of the sufferer. Total incontinence, that is, the continuous loss of urine as opposed to the loss associated with coughing or sneezing, is the most severe type of the malady. For such individuals, the artificial sphincter offers hope for a new life. Incidences of total urinary incontinence as a result of radical prostatectomy in the treatment of carcinoma of the prostate have been reported in the range of 5-50%. Incontinence may occur as a result of injury to the proximal urethra, and it is usually present to some extent in patients with neurogenic bladder dysfunction caused by spinal cord injury, myelomeningocele, or other conditions that affect the micturition centers of the nervous system. Some patients whose urinary tract is completely obstructed and who are therefore unable to urinate, as for example individuals who sustain traumatic complete transection of the urethra with resulting obstructive fibrosis of the urethra, or those patients whose neurogenic spastic sphincter inhibits satisfactory voiding, may benefit from reconstructive surgery or ablation of their pathologic sphincter in order to restore urination. Rehabilitation of such patients can then be complete with implantation of an artificial sphincter to provide urinary control. The alternatives for management include diapers, the placement of external collecting or occlusive devices, or major surgery in which the intestinal tract is used either for conducting the urine to an abdominal collecting bag or as a bladder substitute that is periodically emptied by catheterization.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3050388 TI - Altered vitamin C transport in diabetes mellitus. AB - Tissues sequester vitamin C in concentrations exceeding that present in plasma. The transmembrane transport mechanisms have been shown to be influenced by the concentration of glucose in vitro. On this basis an impairment of tissue vitamin C status may be present in diabetes mellitus. Recent evidence in support of this hypothesis, first proposed by Mann in 1974, is reviewed. PMID- 3050390 TI - What can we learn from the clustering of cases of cancer? PMID- 3050391 TI - The laboratory evaluation of abnormal endocrine function: the fallacy of seeking a single test. PMID- 3050392 TI - The psychosocial consequences of childhood infection with human immunodeficiency virus. AB - This article reviews the available medical literature on the psychosocial and neuropsychiatric implications of childhood infection by the human immunodeficiency virus. This information is supplemented by discussion of cases from our clinical experience. It is clear that different psychosocial issues are confronted by children of different ages (infants, young children or adolescents) and their families. The differences are due partly to the social correlates of different modes of transmission of the acquired immunodeficiency syndrome (AIDS), the psychosocial risk factors that are associated with certain family life styles, whether other family members are infected, and the developmental stage and awareness of the child. Psychiatric assessments of these children and their families are recommended as a routine as they will identify potential ethical dilemmas, and will allow for the detection and management of the psychosocial and neuropsychiatric consequences of AIDS. PMID- 3050394 TI - Alister John Douglas Brass. PMID- 3050393 TI - Therapeutic usage of the non-steroidal anti-inflammatory drugs. PMID- 3050395 TI - The treatment of acne. PMID- 3050396 TI - A perspective on the cost-effectiveness and risks of non-steroidal anti inflammatory drug therapy. PMID- 3050397 TI - Coronary thrombolysis: an important therapy for myocardial infarction. PMID- 3050398 TI - Long-standing fevers in a young pregnant woman. PMID- 3050399 TI - Living, related kidney donors. PMID- 3050400 TI - Breast cancer again: diagnostic alternatives. PMID- 3050401 TI - Retinitis pigmentosa: hope for improved diagnosis and treatment. PMID- 3050403 TI - Accuracy of mammography in an Australian community setting. AB - The records of three surgeons in a Sydney suburban practice have been reviewed to compare the histopathological data from breast biopsies with the clinical assessment, and the mammographic and ultrasound reports, and to assess the extent of the delay before presentation to a surgeon to determine the diagnostic accuracy of the three modalities, and whether mammography contributed to this delay. Two hundred and nineteen breast biopsies that were performed in the years 1984 to 1986 inclusive were reviewed. Of these, 72 were cancerous lesions and 147 were benign lesions. The accuracy (true-positive rate) of mammography for the detection of cancer was 83% with a false-negative rate of 11%. We found no evidence that mammography contributed significantly to the delay before patients presented to specialist surgeons, nor to any delay before they underwent a biopsy. PMID- 3050402 TI - Comparison of mammary serum antigen assay with mammography in patients with breast cancer. AB - Mammary serum antigen levels and two-view xeromammography were evaluated in a study of 97 patients who presented at a specialist breast clinic in a major teaching hospital, in order to determine their single and combined value in the detection of breast cancer. Raised mammary serum antigen levels (greater than 300 inhibition units) were found in 76% of patients with stage-1 (including carcinoma in-situ) and stage-II breast cancer compared with 54% of patients whose mammograms were suggestive of cancer (probability of carcinoma, greater than 50%). Four patients with carcinoma-in-situ had elevated mammary serum antigen levels, but their mammograms did not suggest the presence of cancer (probability of carcinoma, less than 50%). The over-all sensitivity, specificity and predictive value of a positive test-result in detecting breast cancer were 76%, 82% and 72%, respectively, for the mammary serum antigen test, and 54%, 88% and 74%, respectively, for mammography. When the mammary serum antigen test and mammography were used together, a combined evaluation enhanced the sensitivity in the detection of breast cancer (89%) but with a concomitant reduction in specificity (72%). The mammary serum antigen test was superior to mammography in the detection of breast cancer in this study and may prove to be a useful adjunct to conventional methods of clinical assessment and to mammography for the detection of breast cancer. PMID- 3050404 TI - Intravenous anaesthetic agents. AB - Detailed knowledge of the pharmacology of the intravenous anaesthetic agents--a relatively-small group of drugs--is necessary to achieve optimal results in a diverse patient population. The trend towards short-stay surgery requires a consideration of the speed of recovery from anaesthesia, as well as the quality of that recovery, more than ever before. New agents, particularly propofol, have expanded the potential for total intravenous anaesthesia, but technical developments in drug delivery are needed for the full realization of the properties of these drugs. PMID- 3050405 TI - A biological false-positive result for human immunodeficiency virus type-1 in an army recruit. PMID- 3050406 TI - [Bile acids in the blood: a new indicator of hepatic function in occupational exposure to xenobiotics?]. PMID- 3050407 TI - The cervical cap. PMID- 3050408 TI - Prevention of Pneumocystis carinii pneumonia. PMID- 3050409 TI - Extracorporeal photochemotherapy for cutaneous T-cell lymphoma. PMID- 3050410 TI - Naftifine for fungal skin infections. PMID- 3050411 TI - Clomipramine for obsessive compulsive disorder. PMID- 3050412 TI - [Effectiveness of the immunoenzyme method (IEM) for the diagnosis of the early stage of echinococcosis]. PMID- 3050413 TI - [Stimulation of the development of chloroquine-sensitive and -resistant strains of Plasmodium falciparum by cocultivation with Mastomys natalensis cells]. PMID- 3050414 TI - [Cultivation of sandflies of the genus Phlebotomus]. PMID- 3050416 TI - [Effect of food composition on the motility and resorption behavior of the gastrointestinal tract]. PMID- 3050415 TI - [The distribution of echinococcosis in the USSR. Echinococcus multilocularis infection]. PMID- 3050417 TI - [What does "good blood pressure regulation" mean?]. PMID- 3050418 TI - [Intrauterine device--a rare foreign body in the urinary bladder]. PMID- 3050420 TI - [Training and education for the young diabetic and his parents]. PMID- 3050421 TI - [Underwater sports in the developmental years]. PMID- 3050419 TI - Adolescent smoking: research and health policy. AB - A great deal is now known about the prevalence and natural history of adolescent smoking. The literature on smoking intervention, drawn from theoretically based experiments, shows that most programs are unlikely to become widely disseminated because of the extensive resources required. More thoughtful strategies for research and targeting of interventions are examined. They may yield coordinated and comprehensive approaches to moving more quickly and effectively from the behavioral laboratory into the field. PMID- 3050422 TI - [Review of the Pena-Shokeir syndrome I, Pena-Shokeir II and Neu-Laxova. Clinical and interpretative contribution]. PMID- 3050423 TI - [The "asymmetric crying facies": an indication of other malformations?]. PMID- 3050424 TI - [Motilin cells in the intestinal mucosa in celiac disease before, during and after diet therapy. Immunohistochemical study]. PMID- 3050425 TI - [Pelvic echography in the differential diagnosis of isolated precocious thelarche and true precocious puberty]. PMID- 3050426 TI - [Cloperastine fendizoate in the treatment of cough-producing diseases in pediatrics]. PMID- 3050428 TI - [Endemic goiter in Italy: current status]. PMID- 3050427 TI - [Non-insulin-dependent diabetes mellitus in the young]. PMID- 3050429 TI - [Residual beta-cell function in type diabetes]. PMID- 3050430 TI - [Granulosa cell tumor of the hard palate. Case report]. PMID- 3050431 TI - [Treatment of gingival recession with a coronal flap]. PMID- 3050432 TI - [Dental pathology from extra-dental causes. Acquired visceral tooth diseases and prenatal tooth diseases]. PMID- 3050433 TI - [Histiocytosis of the Langerhans cells. Etiopathogenesis and prognostic picture in the light of the most recent findings]. PMID- 3050434 TI - [Sarcomas of the jaws]. PMID- 3050436 TI - [Bacterial recolonization of deep periodontal lesions after nonsurgical therapy]. PMID- 3050435 TI - [Subgingival anaerobic flora in periodontal disease in adults]. PMID- 3050437 TI - [Diagnosis and therapy of benign neoplasms of the dental system and jaw bones]. PMID- 3050440 TI - [Unicystic ameloblastoma and plexiform epithelial hyperplasia. I. Histogenetic aspects]. PMID- 3050438 TI - [Indications and comparative evaluation of chlorhexidine in periodontal therapy]. PMID- 3050439 TI - [Comparative radiographic and histopathologic study on tissue response to endodontic materials in the rabbit femur]. PMID- 3050441 TI - [Dental anomalies subsequent to pathogenic noxae and/or noxae associated with genetic factors]. PMID- 3050442 TI - [Prevention of AIDS in dental practice]. PMID- 3050443 TI - [Use of thymopentin in the therapy of recurrent oral ulcers]. PMID- 3050444 TI - Nonimmune hydrops fetalis. PMID- 3050445 TI - Medicaid expands to cover more pregnant women & children. PMID- 3050446 TI - Transducin inhibition of light-dependent rhodopsin phosphorylation: evidence for beta gamma subunit interaction with rhodopsin. AB - Rhodopsin kinase was purified from bovine retina rod outer segments as a 62-64 kDa protein that phosphorylated purified rhodopsin reconstituted into egg phosphatidylcholine/phosphatidylethanolamine liposomes. A competition binding assay in which transducin competes with rhodopsin kinase for binding sites on rhodopsin was used to assess the interaction of purified transducin subunits with rhodopsin. Preincubation of purified holotransducin with rhodopsin, in the absence of guanosine triphosphate, blocked the ability of the kinase to phosphorylate rhodopsin. Transducin-dependent inhibition of phosphorylation was relieved when guanosine 5'-(3-O-thio)triphosphate was present during the preincubation. Resolved alpha and beta gamma transducin subunits, in the absence of guanosine triphosphate, were each capable of specifically blocking phosphorylation of rhodopsin. A maximally effective concentration of T alpha or T beta gamma (1 microM) subunits inhibited phosphorylation of rhodopsin (0.23 microM) 45-65%. A similar concentration of reconstituted transductin (T alpha and T beta gamma) or native holotransducin (T alpha beta gamma) inhibited phosphorylation greater than 98%. The results indicate that rhodopsin must have a binding site for T beta gamma as well as a binding site for T alpha, and each subunit influences the recognition of bleached rhodopsin by rhodopsin kinase. PMID- 3050447 TI - Interaction of 2-halogenated dATP analogs (F, Cl, and Br) with human DNA polymerases, DNA primase, and ribonucleotide reductase. AB - Recently, 2-halogenated deoxyadenosine analogs (F, Cl, and Br) have been shown to have antitumor activity. These analogs are phosphorylated by cells and are believed to exert their cytotoxic action at the nucleoside triphosphate level. In this work the interaction of these nucleoside triphosphate analogs with potential targets, such as DNA polymerase alpha, beta, and gamma, DNA primase, and ribonucleotide reductase was examined in detail. All of these compounds competitively inhibited the incorporation of dAMP into DNA by DNA polymerase alpha, beta, or gamma. F-dATP was able to completely substitute for dATP using DNA polymerase alpha and gamma, but not with DNA polymerase beta. Cl-dATP and Br dATP substituted poorly for dATP using DNA polymerase alpha and beta. Extension of a 32P-labeled primer by DNA polymerase alpha, beta, or gamma on a single stranded M13 template showed that these compounds were incorporated into the 3' end of the growing DNA chain and that elongation beyond the incorporated analogs was significantly retarded for Cl-dATP and Br-dATP using either DNA polymerase alpha or beta. DNA primase using poly(dC) as template was inhibited by these compounds at a concentration 4 to 5 times greater than that required for 2-F araATP. The 2-halogenated dATP analogs were potent inhibitors of ADP reduction by ribonucleotide reductase. In conclusion, the cytotoxic action of 2-Cl deoxyadenosine and 2-Br-deoxyadenosine may partially be mediated through the mechanism of "self-potentiation," by depression of the deoxynucleoside triphosphate pools due to inhibition of ribonucleotide reductase, which would facilitate their incorporation into DNA and result in the inhibition of DNA synthesis. PMID- 3050450 TI - Control of mitochondrial respiration in muscle. AB - Control of mitochondrial respiration depends on ADP availability to the F1 ATPase. An electrochemical gradient of ADP and ATP across the mitochondrial inner membrane is maintained by the adenine nucleotide translocase which provides ADP to the matrix for ATP synthesis and ATP for energy-dependent processes in the cytosol. Mitochondrial respiration is responsive to the cytosolic phosphorylation potential, ATP/ADP.Pi which is in apparent equilibrium with the first two sites in the electron transport chain. Conventional measures of free adenine nucleotides is a confounding issue in determining cytosolic and mitochondrial phosphorylation potentials. The advent of phosphorus-31 nuclear magnetic resonance (P-31 NMR) allows the determination of intracellular free concentrations of ATP, creatine-P and Pi in perfused muscle in situ. In the glucose-perfused heart, there is an absence of correlation between the cytosolic phosphorylation potential as determined by P-31 NMR and cardiac oxygen consumption over a range of work loads. These data suggest that contractile work leads to increased generation of mitochondrial NADH so that ATP production keeps pace with myosin ATPase activity. The mechanism of increased ATP synthesis is referred to as 'stimulus-response-metabolism' coupling. In muscle, increased contractility is a result of interventions which increase cytosolic free Ca2+ concentrations. The Ca2+ signal thus generated increases glycogen breakdown and myosin ATPase in the cytosol. This signal is concomitantly transmitted to the mitochondria which respond to small increases in matrix Ca2+ by activation of Ca2+-sensitive dehydrogenases. The Ca2+-activated dehydrogenase activities are key rate-controlling enzymes in tricarboxylic acid cycle flux, and their activation by Ca2+ leads to increased pyridine nucleotide reduction and oxidative phosphorylation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3050451 TI - Antigenic homology between rat sperm tail polypeptides and Sertoli cell secretory proteins. AB - A high performance liquid chromatographic procedure has been used for the purification of rat Sertoli cell secretory protein S70 and S45-S35 heterodimeric protein to determine their role during spermatogenesis. These two proteins display binding affinity for each other and appear antigenically related. We have observed that: 1. S70 and S45-S35 heterodimeric protein coelute during purification, 2. polyclonal antiserum raised against protein S70 recognizes common antigenic determinants in polypeptides S45 and S35, the disulfide-linked components of the heterodimeric protein, and 3. a monoclonal antibody that recognizes polypeptide S35 but does not crossreact with either protein S70 or polypeptide S45, immunoprecipitates the S70/S45-S35 heterodimeric protein complex. In immunofluorescent experiments, antisera raised against protein S70 and polypeptide components of S45-S35 heterodimeric protein immunoreact with two major sperm intracellular structures: the acrosome and periaxonemal outer dense fibers of sperm tail. Immunoreactivity was not detected on the sperm plasma membrane surface of unfixed, living sperm. Outer dense fibers extracted from sperm tails by a combined treatment with cetylthrimethylammonium bromide and 2 mercaptoethanol, yielded a characteristic polypeptide pattern. In immunoblotting experiments sperm tail polypeptides were recognized by polyclonal antisera raised against Sertoli cell secretory proteins. We conclude that Sertoli cell secretory proteins S70 and S45-S35 heterodimeric protein are antigenically related to each other and to keratin-like polypeptides from sperm tail. PMID- 3050449 TI - On the nature of the Duchenne muscular dystrophy locus: a portion of a complex of related gene clusters of recent pseudoautosomal origin? AB - The current hypothesis that the Duchenne/Becker muscular dystrophy locus encodes a single 2,000 kb gene is analyzed. The apparent encoding efficiency, the individual and total exon/intron ratios, and the heterogeneity of deletions associated with the disease, which are currently interpreted as supporting the single gene hypothesis, are also consistent with the alternative hypothesis that this locus is a portion of a complex of related gene clusters which include synthenic transcriptional units of enzymes and ligand transport proteins of one or more convergent metabolic pathways. The high recombination frequency and high rate of deletions are consistent with a locus that has recently evolved from pseudoautosomal origin. The propositions that nebulin or dystrophin is the product of the DMD locus, and that the mdx locus in the mouse is homologous to that of DMD, are critically evaluated. Several lines of evidence support the contention that developmental and tissue-specific enzymes of acyl-specific phospholipid synthesis are encoded in these clusters. Phenotypic variability not accountable for by deletion heterogeneity is postulated to arise from epistatic interactions with other loci within or outside these putative clusters. Some testable predictions of these hypotheses are suggested. PMID- 3050453 TI - DNA hybridization with oligodeoxyribonucleotide probes for identifying enterotoxin-producing Escherichia coli. AB - The DNA hybridization method using oligodeoxyribonucleotide probes was compared with the GM, ELISA and the infant mouse assay for determining production of LT and ST, respectively, by 2000 strains of Escherichia coli. Sensitivity and specificity by DNA hybridization for LT were 98.7 and 99.8%, and for ST were 78.8 and 99.6%. PMID- 3050452 TI - Botulinum neurotoxin types A, B & E: pH induced difference spectra. AB - The alkaline pH induced difference spectra (270-310 nm) of three antigenically distinct forms of the botulinum neurotoxin (NT) types A, B and E were examined. When isolated from the cultures of Clostridium botulinum, type A NT is a fully toxic dichain (nicked) protein, type E is a mildly toxic single chain (unnicked) protein, and type B NT is a mixture of single and dichain proteins and near fully toxic. Trypsin nicks the single chain protein to the dichain and increases its toxicity (up to about 100 fold in type E). A strong difference spectrum peak at approximately 296 nm was found when types A, B or E NT were in the alkaline pH region. This peak was not observed at pH 4.0. For types A and B NT plots of difference absorptivity vs. pH were simple sigmoidal curves. The pK of phenolic moieties of tyrosine residues in both proteins were 10.9. Nearly all tyrosine residues in both proteins were ionized. The single chain type E, unlike type A and B NT, yielded a two step titration curve and pK values 11.3 and less than 7.5; about 60% of the total tyrosine residues present were ionized. The two step titration curve was not observed when the single chain protein was nicked with trypsin to the dichain type E NT. The titration curve of dichain type E NT, although complex, was more like those of type A and B NT. PMID- 3050448 TI - The metabolic environment of cancer. AB - The tumor cell has a very distinctive metabolism. It acts as a metabolic trap for host nutrients thus taking vital compounds for the metabolism of the host. Depending on the particular tumor growing pattern, cancer cells use preferentially glucose or amino acids for their energetic or biosynthetic needs. Lipids, fatty acids in particular, can also be taken up by the tumor cell. In addition, it can also release some compounds into the host circulation which are not normally produced by the original cell before neoplastic transformation. Some of these compounds affect the metabolism of the host in an unfavorable way since they can oppose the host's metabolic responses, which sustain homeostasis. The final product is that the metabolic machinery of these cells allows them to grow continuously in an uncontrolled manner. The consequences of tumor invasion on the host's metabolism are varied. They have, however, one thing in common: the reduction of the metabolic efficiency of the host. Muscular protein depletion, increased gluconeogenesis, uncoupling of oxidative phosphorylation constitute the main metabolic responses of the host as a result of tumor invasion. The net result of all these metabolic changes is profound energy imbalance which normally ends with cachexia and, eventually, death. PMID- 3050454 TI - Nucleic acids or immunoglobulins: which are the molecular probes of the future? PMID- 3050455 TI - Experimental autoimmune myasthenia gravis. PMID- 3050457 TI - Molecular dissection of the nicotinic acetylcholine receptor. PMID- 3050456 TI - Myasthenia gravis: new therapeutic strategies. PMID- 3050459 TI - Pathobiology and recognition of malignant melanoma: Introduction. PMID- 3050460 TI - Dysplastic nevus: a formal histogenetic precursor of malignant melanoma. PMID- 3050458 TI - Antibody specificity in myasthenia gravis. PMID- 3050462 TI - Melanocytic proliferations that simulate malignant melanoma histopathologically. PMID- 3050463 TI - Biological and prognostic significance of vertical growth phase characteristics in malignant melanoma. PMID- 3050461 TI - Minimal deviation melanoma. PMID- 3050464 TI - Melanoma markers: biological and diagnostic considerations. PMID- 3050465 TI - Immunology of melanoma. PMID- 3050466 TI - Histologic reporting of malignant melanoma. AB - The schema for histologic reporting of malignant melanoma outlined above is incomplete. A number of variables, including quantification of tumor cell pigmentation, description of solar degeneration in surrounding stroma, and extent of S-100 protein immunoreactivity by tumor cells, have not been addressed. In addition, it is likely that within the next several years, additional parameters, including those identified by the use of in situ genomic probes (Chapter 3), will become known to confer prognostic information when assessed histologically in biopsy material. Nonetheless, this approach represents a start, an attempt to achieve uniformity in histologic reporting of melanoma. It represents a three tiered approach. First is the establishment of a malignant melanocytic lesion (malignant melanoma) and the assessment of the anatomic level and extent of invasion of the dermis, which correlates directly with metastatic potential (the vertical growth phase measured in millimeters). Subclassification of the radial growth phase is optional and, by convention, is also reported in this "above the line" portion of the diagnostic assessment. Second, a number of subsidiary diagnostic statements, usually in the form of notes or comments, are recommended. These include assessment of overlying epidermal ulceration, a description of the morphology and mitotic activity of the vertical growth phase nodule, an assessment of the presence or absence of vascular invasion and microscopic satellite formation, a description of the host mononuclear cell response and/or regression, and mention of associated pathology. Finally, an additional narrative statement may be included to describe such things as extent of S-100 immunoreactivity or to modify in a more descriptive manner the database provided above. Although this last category is neither crucial nor mandatory, it serves to add to a collective database from which future determinations concerning the prognostic significance of new histologic parameters may be formulated. Table 6.3 provides a sample report based on the above considerations. This method of histologic reporting is recommended because it includes important diagnostic information as well as parameters for prognosis. Although certain details of less impelling current significance are included, these are of potential future importance as a greater number of patients with malignant melanoma are followed prospectively for prolonged periods of time. These factors may also prove to be significant in the accumulation of a database for research. PMID- 3050467 TI - Problems in microstaging of melanoma vertical growth. AB - Review of the currently available data would indicate that the measurement of tumor thickness in millimeters, especially those greater than 1.5 mm in thickness, is the single most important prognostic variable, closely followed by sex and location of the tumor. On the other hand, in thinner tumors, the level of invasion takes precedence over tumor thickness, especially in the setting of the "thin level IV" tumor. The microstaging must therefore include measurements of both level and thickness (keeping in mind the problems involved in thickness measurements noted above), as well as notation of special features of prognostic value (mitotic index, ulceration, microsatellites, etc.). Only with such refinements and the continued study of parameters from large databases can the estimates of prognosis in patients with primary malignant melanoma be made sufficiently accurate to be useful in planning appropriate therapy. PMID- 3050468 TI - Wilson's disease: yesterday, today, and tomorrow. PMID- 3050469 TI - Remembering Kinnier Wilson. PMID- 3050470 TI - Trauma as an etiology of parkinsonism: a historical review of the concept. AB - The relationship between trauma and the development of Parkinson's disease has been an issue in neurology since James Parkinson's initial 1817 essay. This paper will delineate the historical development of the concept of trauma as an etiologic agent in Parkinson's disease and parkinsonism. The strong influence of socioeconomic societal forces in the 1870s with regard to liability laws and the subsequent acceptance in the medical community of the role of peripheral trauma in producing Parkinson's disease is presented. We will also review present-day criteria for assigning head trauma an etiologic role in parkinsonism. The discussion will stress current knowledge regarding trauma and parkinsonism, and it will also review the issues of the possible role of the current litigious society's influence on determining a role for trauma in Parkinson's disease. PMID- 3050471 TI - Madopar HBS in fluctuating parkinsonian patients: two-year treatment. AB - In an open-label study, we substituted conventional levodopa plus benserazide: 100/25 (Madopar) with a controlled-release form (HBS) in 18 fluctuating parkinsonian patients for 24 months. Significantly positive results were obtained in both peak-dose and diphasic dyskinesias up to 12 months of treatment; morning akinesias were also improved up to 6 months. A general trend of deterioration, compared to the first 3-6 months of HBS treatment, was observed in "off" fluctuations after 1 year: akinesias due to a delayed response worsened after 1 year of treatment also when compared with the conventional treatment. Positive results were obtained with new HBS on standard Madopar-related psychiatric disorders. PMID- 3050473 TI - Memories of my father [Kinnier Wilson]. PMID- 3050472 TI - Analysis of open-label trials in torsion dystonia using high dosages of anticholinergics and other drugs. AB - We reviewed the records of 358 patients with various forms of focal, segmental, and generalized dystonia who had received pharmacotherapy in a systematic order, beginning with anticholinergics. If no benefit was encountered, we then tested clonazepam, baclofen, and other agents sequentially. In each situation the dosage was gradually increased until benefit or troublesome adverse effects were encountered. In this manner we obtained data on the percentage of patients who showed moderate to marked benefit. Anticholinergics were the most beneficial agent, confirming previous reports. Statistical analysis of this response revealed that benefit from anticholinergics was most likely if treatment was begun within 5 years after the onset of dystonia. From this analysis, it appears that delaying treatment beyond 5 years is likely to result in an unsatisfactory response. PMID- 3050474 TI - Hemiballism in multiple sclerosis. AB - Hemiballism has rarely been reported in the setting of multiple sclerosis. Review of clinical descriptions cast some doubt on the accuracy of the diagnosis of the movement disorder in several previous cases. We report a patient with definite multiple sclerosis, with videotape documentation of hemiballism, and demonstration of a plaque in the contralateral subthalamic nucleus on a magnetic resonance image. PMID- 3050475 TI - Some immunological properties of high and low tumorigenic cellular variants of c H-ras transformed 3T3 cells. AB - NIH 3T3 cells transformed in vitro with the c-H-ras oncogene were subcloned. The resulting subclones were assayed for in vivo tumorigenicity in nude and in immunocompetent mice. The response of two high tumorigenic and two low tumorigenic clones to mediators of natural immunity was analyzed. The clones did not differ in sensitivity to NK cell-mediated lysis. However, compared to low tumorigenic clones, the high tumorigenic ones had a down-regulated expression of a membrane determinant recognized by a certain monoclonal naturally occurring antibody. The determinants recognized by other monoclonal naturally occurring antibodies available in the laboratory were equally expressed on the high and low tumorigenic clones. The high tumorigenic cells showed an increased resistance to cytotoxicity mediated by lymphotoxin. These results suggest that naturally occurring antibodies and lymphotoxin may participate in controlling the tumorigenicity of transformed cells. The high tumorigenic clones but not the low tumorigenic ones contained a novel 3.5-kb ras mRNA. PMID- 3050476 TI - Adoptive immunotherapy in conjunction with bone marrow transplantation- amplification of natural host defence mechanisms against cancer by recombinant IL 2. PMID- 3050477 TI - Prevention and treatment of kidney stones. PMID- 3050478 TI - [Comparative electron microscope studies in collagenous colitis and in the fibrotic stage of chronic ulcerative colitis]. PMID- 3050479 TI - [Changes in the endocrine pancreas in chronic pancreatitis]. PMID- 3050480 TI - [Etiology, clinical picture and forensic assessment of the battered child syndrome. II]. PMID- 3050482 TI - [Possible determination of cell cycle parameters by Feulgen-DNA cytophotometry from cell nuclei isolated from paraffin-embedded tumor tissue]. PMID- 3050481 TI - [Lung diseases caused by animal allergens]. PMID- 3050483 TI - [Structural rearrangements of the plasmid R68,45 in cells of Brucella suis]. AB - The mobilizing activity of the plasmid R68,45 in Escherichia coli and Brucella abortus has been studied. The plasmid R68,45 has been found to lose its ability to mobilize the chromosome in Brucella suis cells. The experiments on the conjugational transfer of R68,45 were confirmed by restriction analysis of the plasmid DNA. R68,45 has been shown to lose Cma in brucella cells via deletion. The molecular mechanisms of deletion process in brucella and other bacteria are discussed. PMID- 3050484 TI - [Genetics of susceptibility to alcoholism]. PMID- 3050486 TI - Kinetochores, centromeres, spindles and the induction of aneuploidy. PMID- 3050485 TI - [Cloning and expression of the histidine operon of Salmonella typhimurium in cells of Escherichia coli]. AB - The histidine operon of Salmonella typhimurium and its fragments were cloned in Escherichia coli cells on a multicopy plasmid. Expression of the cloned genes and histidine production by the variants possessing the hisG mutation which desensibilizes the ATP phosphoribosyl transferase for histidine were studied. Amplification of the complete operon including the hisG gene enables histidine accumulation of 2-3 g/l after 72 hours of fermentation. PMID- 3050487 TI - Studies of the interaction of chemicals with microtubule assembly in vitro can be used as an assay for detection of cytotoxic chemicals and possible inducers of aneuploidy. PMID- 3050488 TI - Cytogenetic activities of tobacco particulate matter (TPM) derived from a low to middle tar British cigarette. AB - Tobacco particulate matter (TPM) derived from an experimental low to middle tar cigarette was tested for its cytogenetic activity upon a low passage number Chinese hamster pulmonary cell line. Examination of the mitotic profiles (after one cell cycle) revealed no interference by the agent with mitotic spindle formation and/or function. However, complete chromosomes (or parts of them) were seen to dislocate from the mitotic spindles. Such an event was probably the result of the chromosome aberrations, substantial numbers of which were observed in second division cells, or through a process of centromere inactivation. In second division cells there was a reduction in the number of diploid cells accompanied by an increase in both hypodiploidy and polyploidy and there was also a non-dose-related increase in endoreduplication. The results demonstrate that TPM was capable of inducing both structural and numerical chromosome aberrations in cultured mammalian cells. PMID- 3050489 TI - Induction of meiotic chromosomal malsegregation in yeast. AB - A system to detect chromosome number abnormalities occurring during meiosis in Saccharomyces cerevisiae is described. It is based on selection of spores carrying 2 multi-marked chromosomes V. Each step of the technical procedure is critically analyzed and the origin of some biases discussed. Selection and subsequent genetic analysis allow the estimation of the frequency of spontaneous and induced diploid and aneuploid n + 1 (diplo-V) spores. Data are reported concerning the effect of 53 chemical compounds. The great majority of active chemicals induce diploid clones while a minority cause non-disjunction of chromosome V. PMID- 3050490 TI - Induction of chromosome malsegregation by halogenated organic solvents in Aspergillus nidulans: unspecific or specific mechanism? AB - Three chloromethanes (dichloromethane, chloroform and carbon tetrachloride) and 8 chlorinated ethanes (1,1- and 1,2-dichloroethane, 1,1,1- and 1,1,2 trichloroethane, 1,1,1,2- and 1,1,2,2-tetrachloroethane, pentachloroethane and hexachloroethane) were assayed in tests for the induction of mitotic segregation in Aspergillus nidulans diploid strain P1. Eight of the 11 compounds assayed (dichloromethane, chloroform, carbon tetrachloride, 1,1- and 1,2-dichloroethane, 1,1,2-trichloroethane, 1,1,1,2- and 1,1,2,2-tetrachloroethane) significantly increased the frequency of morphologically abnormal colonies which produced euploid whole-chromosome segregants (haploids and non-disjunctional diploids). Only in one case (1,1,1,2-tetrachloroethane) was a borderline increase in crossing-over frequency observed, thus suggesting the involvement of non-DNA targets in aneuploidy induction by these chlorinated hydrocarbons. Conclusive evidence for the induction of aneuploidy as the primary genetic event was provided by experiments in haploid strain 35 with 1,2-dichloroethane and 1,1,1,2 tetrachloroethane. Mutagenic, lethal and growth-arresting activities were quantitatively estimated and compared to a series of descriptors of physical and chemical properties of the molecules by means of multivariate statistical analysis. Lipophilicity, known to be related to c-mitotic activity, did not show any significant relationship with aneuploidizing activity, whereas a possible correlation among physico-chemical descriptors and toxic properties of test chemicals was highlighted. PMID- 3050491 TI - Investigations of aneuploidy-inducing chemical combinations in Saccharomyces cerevisiae. AB - For several years we have been investigating combinations of chemicals for their ability to induce aneuploidy. Earlier published results indicated that combinations of certain chemicals showed a potentiation effect while other combinations did not. We have continued to explore this phenomenon and report additional findings in this communication. Combinations of ethyl acetate and methyl ethyl ketone showed a potentiation effect as did 1-methyl-2-pyrrolidinone nocodazole combinations. Combinations that did not show a potentiation effect were 2-pyrrolidinone-nocodazole and 1-methyl-2-pyrrolidinone-ethyl acetate. We also found that nocodazole, which is a potent inducer of aneuploidy in yeast extract-peptone-dextrose (YEPD) medium but not in synthetic complete (SC) medium, showed a potentiation effect with ethyl acetate in SC medium. This effect in SC medium is similar to that previously reported for nocodazole with ethyl acetate in YEPD medium. When nocodazole was dissolved in 1-methyl-2-pyrrolidinone as a concentrated stock solution, a potentiation effect occurred even at low concentrations of the solvent. PMID- 3050492 TI - A genetic assay for aneuploidy: quantitation of chromosome loss using a mouse/human monochromosomal hybrid cell line. AB - A genetic assay is described in which a mouse/human hybrid cell line R3-5 containing a single human chromosome (a monochromosomal hybrid) is used to detect chemically induced aneuploidy. In this assay the frequency of chromosome loss determined by the cloning efficiency of the cells in a selection medium is used as an index for the potential of a chemical to induce aneuploidy. The hybrid cells are deficient in hypoxanthine guanine phosphoribosyltransferase (HGPRT) and contain human chromosome 2, marked with Ecogpt, an E. coli gene for xanthine guanine phosphoribosyltransferase. These cells with a genotype of hgprt-/Ecogpt+ can grow in medium containing mycophenolic acid and xanthine (MX medium) but not in medium containing 6-thioguanine (6-TG). The loss of the human chromosome from R3-5 cells as a result of chemical treatment produces cells with a genotype of hgprt-/Ecogpt- which are capable of growth in the medium containing 6-TG. Thus, the cloning efficiency of cells treated with a test chemical in 6-TG provides a method to determine the frequency of cells that have lost the human chromosome. Two chemicals, colcemid and nocodazole, previously known to induce aneuploidy in mammalian cells were used for a preliminary evaluation of this test system. Both of these compounds at concentrations ranging from 0.002 to 0.032 micrograms/ml showed a concentration-related positive response in this assay. PMID- 3050493 TI - Aprotic polar solvents that affect porcine brain tubulin aggregation in vitro induce aneuploidy in yeast cells growing at low temperatures. AB - Seven aprotic polar solvents which had previously been shown to interfere with the aggregation in vitro of porcine brain tubulin have been examined for their ability to induce mitotic aneuploidy in Saccharomyces cerevisiae in relation to temperature during exposure. Induction of aneuploidy was in general considerably enhanced when incubation at 28 degrees C was interrupted by overnight storage at low temperature (cold shock). The optimum cold-shock temperatures for individual chemicals varied over a range of 0-16 degrees C. While storage at reduced temperature enhanced the effect of treatment at 28 degrees C, it was also shown that continuous incubation at reduced temperature could greatly enhance the induction of aneuploidy. Only 2 chemicals, 1-methyl-2-pyrrolidinone and gamma valerolactone, required cold shock to yield a positive response. The other chemicals did not require cold shock for enhanced induction. The observation that the agents examined also interfere with in vitro tubulin aggregation suggests that there is a temperature component to the interaction of these agents with tubulin in vivo. This temperature component is unusual in that the most effective temperature range for aneuploidy induction can be well below the optimal growth temperature for the test organism. PMID- 3050494 TI - Immunofluorescent staining of kinetochores in micronuclei: a new assay for the detection of aneuploidy. AB - The immunofluorescent staining of kinetochores in micronuclei with antikinetochore antibodies was used to develop an in vitro assay for aneuploidy inducing agents. The results show that about 80% of micronuclei induced by either colchicine or chloral hydrate contained kinetochores; only 9% of X-ray-induced micronuclei reacted positively to the antibody. These findings indicate that the in vitro micronucleus assay coupled with immunofluorescent staining of kinetochores can be a useful method for assessing the ability of chemicals to induce aneuploidy and/or chromosome aberrations. PMID- 3050495 TI - Mutagenic activities of selected nitrofluoranthene derivatives in Salmonella typhimurium strains TA98, TA98NR and TA98/1,8-DNP6. AB - The mutagenic activities of novel nitrofluoranthene derivatives in Salmonella strains TA98, TA98NR and TA98/1,8-DNP6 (with and without S9 addition) are given. These derivatives were produced from the reactions of fluoranthene (FL) and its directly mutagenic 2- and 3-nitro derivatives with covalent dinitrogen pentoxide (N2O5) in CCl4 solution at ambient temperature. The influence of the addition of a nitro group on the observed activity of the resulting di- and tri nitrofluoranthenes is discussed. PMID- 3050496 TI - The use of rat urine extracted on XAD-2 resin and assayed in a microsuspension modified Ames test as an in vivo indicator of genotoxic exposure. AB - The measurement of urinary mutagenicity is a non-invasive monitoring tool which often reflects an animal's recent exposure to genotoxic agents. Although studies in man are indispensable for monitoring industrial and/or environmental exposure to genotoxins, a sensitive laboratory animal model is necessary for mechanistic studies on the role of specific chemical exposure in altering urinary mutagenicity. The objective of this study was to enhance the sensitivity of the methodology used for detecting urinary mutagenicity in rats by using XAD-2 resin to extract and concentrate the urine and a microsuspension-modified Ames test to quantify mutagenicity. The polycyclic aromatic hydrocarbon benzo[a]pyrene (BP) and the aromatic amine 2-acetylaminofluorene (AAF) were used as test compounds. Under the conditions of our study, AAF administered to rats by gavage at doses of 1 mg/kg or higher induced a dose-dependent increase in urine mutagenicity. The greatest mutagenic response was seen when S9 was present during the microsuspension-modified Ames test and beta-glucuronidase (BG) was not included. Similarly, BP administered to rats by gavage at doses of 10 mg/kg or higher induced a dose-dependent increase in urinary mutagenicity. The relative importance of BG and S9 were quite different with BP than with AAF. With BP, mutagenicity was greatest when both S9 and BG were present during the microsuspension-modified Ames test, and least with S9 and without BG. In both AAF and BP-treated animals, extraction of the urine on XAD-2 resin markedly enhanced the mutagenic response compared to neat urine, but partitioning of the XAD-2 eluate into methylene chloride always diminished the mutagenicity of the urine extract. The results demonstrate the sensitivity and reproducibility of rodent urinary mutagenicity assays when XAD-2 resin is used to extract and concentrate the urine and a microsuspension-modified Ames test is used to quantify mutagenicity. This sensitive method should facilitate mechanistic studies on the roles of specific environmental agents in affecting urinary mutagenicity and, in addition, may be used during acute, subchronic and chronic rodent bioassays as a non-invasive in vivo indicator of genotoxic exposure. PMID- 3050497 TI - Mutagenicity studies on ketone solvents: methyl ethyl ketone, methyl isobutyl ketone, and isophorone. AB - 3 ketone solvents (methyl ethyl ketone (MEK), methyl isobutyl ketone (MiBK), and isophorone) were tested for potential genotoxicity. The assays of MEK and MiBK included the Salmonella/microsome (Ames) assay, L5178Y/TK+/- mouse lymphoma (ML) assay, BALB/3T3 cell transformation (CT) assay, unscheduled DNA synthesis (UDS) assay, and micronucleus (MN) assay. Only the ML, UDS, and MN assays were conducted on samples of isophorone. No genotoxicity was found for MEK or isophorone. The presence of a marginal response only at the highest, cytotoxic concentration tested in the ML assay, the lack of reproducibility in the CT assay, and clearly negative results in the Ames assay, UDS and MN assays, suggest that MiBK is unlikely to be genotoxic in mammalian systems. PMID- 3050498 TI - Ames mutagenicity and concentration of the strong mutagen 3-chloro-4 (dichloromethyl)-5-hydroxy-2(5H)-furanone and of its geometric isomer E-2-chloro 3-(dichloromethyl)-4-oxo-butenoic acid in chlorine-treated tap waters. AB - The Ames mutagenicity and the concentration of the strong Ames mutagen 3-chloro-4 (dichloro-methyl)-5-hydroxy-2(5H)-furanone (MX) and its geometric isomer E-2 chloro-3-(dichloromethyl)-4-oxobutenoic acid (E-MX), derived from chlorination of humus, were determined in XAD extracts of tap water collected from 26 localities in Finland. The 23 tap waters treated with disinfectants gave a positive response in strain TA100. MX and E-MX were detected in all extracts exhibiting mutagenicity with the exception of 3 extracts of marginal activity. MX accounted for 15-57% (average 33%) of the observed mutagenicity. The concentration of E-MX was slightly lower than the corresponding concentration of MX. Linear correlations were observed between mutagenicity and concentration of MX and E-MX, with correlation coefficients of 0.894 for MX and 0.910 for E-MX. PMID- 3050499 TI - Study on the mutagenicity of nifurtimox and eight derivatives with the L arabinose resistance test of Salmonella typhimurium. AB - The mutagenicity of nifurtimox (nfx) and 8 nfx analogues has been investigated with the L-arabinose forward-mutation assay of Salmonella typhimurium. The nfx analogues tested were obtained by replacing the 3-methyl-4-yl-tetrahydro-1,4 thiazine-1,1-dioxide group of the parent compound with the following other groups: indazol-1-yl (1); pyrazol-1-yl (2); benzimidazol-1-yl (3); 1,2,4-triazol 4-yl (4); 1-methyl-3-methylthio-1,2,4-triazol-4-yl-5-thione (5); 3,5 bis(methylthio)-1,2,4-triazol-4-yl (6); 1-adamantyl (7); 4,6-diphenylpyridin-1-yl 2-one (8). The mutagenic activity of each chemical was determined by the standard plate-incorporation test, in the presence or absence of the S9 activation mixture. The 9 compounds were mutagenic and exhibited linear dose-mutagenic response relationships. They were direct-acting mutagens and showed a nearly 1000 fold range in mutagenic potency from chemical 1 to nfx. In most cases, the addition of S9 mixture to the test plates decreased the mutagenicity of compounds. This effect was particularly noticeable in the case of chemicals 1-3, 5 and 7 where a more than 70% decrease in mutagenic activity was observed in the presence of the S9 mixture. The mutagenic potency of compounds in the Ara test showed a negative linear correlation with previously reported antitrypanosomal activity. Thus, chemicals 6 and 8 with in vitro activities against Trypanosoma cruzi clearly superior to that of nfx showed 2 of the lowest mutagenic potencies in the Ara test and these were only somewhat higher than the mutagenicity of the reference drug. PMID- 3050500 TI - The mutagenic potencies of plant extracts containing quercetin in Salmonella typhimurium TA98 and TA100. AB - Four commercial ethanolic plant extracts, Tinctura Alchemillae, Extractum Crataegi, Extractum Myrtilli and Tinctura Hyperici, were tested for their mutagenicity in Salmonella typhimurium TA98 and TA100 with and without S9 mix obtained from rats pretreated with phenobarbital. The extracts studied differed greatly in their mutagenic potencies but exhibited a very similar mutation pattern in which the strongest effect was always seen in tester strain TA98 with S9 mix. Simultaneously we investigated the extracts for the presence of quercetin and kaempferol. Only quercetin was detected in small amounts by thin-layer chromatography (TLC). The fractions containing quercetin were separated and collected using a Sephadex LH-20 column. Two different methods were employed to estimate the amount of quercetin in the extracts: a colorimetric assay developed by Christ and Muller, and a complexometric method by Belikov. The quercetin concentrations ranged between 2 mg (Tinctura Alchemilla) and 89 mg (Tinctura Hyperici) per 100 g of extract. We suggest that the mutagenicity of the 4 plant extracts is mainly due to the presence of quercetin for the following reasons: (1) all the plant extracts exhibit a mutation pattern which is very similar to that of quercetin, (2) the mutagenic potential of the extracts correlates well with their quercetin content, considering the fact that plant extracts are very complex mixtures often containing toxic or antimutagenic compounds. PMID- 3050501 TI - Genetic effects of chlorinated ethylenes in the yeast Saccharomyces cerevisiae. AB - The chlorinated ethylenes 1,1-dichloroethylene (vinylidene chloride), trans-1,2 dichloroethylene, trichloroethylene, and tetrachloroethylene (perchloroethylene) were assayed for their ability to induce mitotic gene conversion and point mutation as well as mitotic aneuploidy in diploid strains of the yeast Saccharomyces cerevisiae. From strain D7 late logarithmic-phase cells grown in 20% glucose liquid medium, containing a high level of cytochrome P-450, as well as stationary-phase cells combined with an exogenous metabolic activating system (S9) were used, in order to activate the chlorinated compounds and to produce electrophilic mutagenic intermediates. Only 1,1-dichloroethylene exhibited a dose dependent genetic activity, while the other ethylenes did not. The 2 ways of metabolic activation were compared and were found to cause approximately the same effect. In contrast to the findings with strain D7, vinylidene chloride, trans 1,2-dichloroethylene, and trichloroethylene induced, without metabolic activation, mitotic chromosomal malsegregation in strain D61.M. The presence of liver homogenate as an activating system did not enhance the respective frequencies of chromosome loss. In the case of tetrachloroethylene, sufficient data have not become available, since this compound showed a highly toxic effect towards yeast cells, decreasing the rate of surviving cells to less than 30% at a concentration of 9.8 mM. PMID- 3050502 TI - Antimutagenic effects of several subfractions of extract from wheat sprout toward benzo[a]pyrene-induced mutagenicity in strain TA98 of Salmonella typhimurium. AB - The aqueous extract from wheat sprouts contains some antimutagenic factor(s). The factor(s) abolish(es) the activity of aryl hydrocarbon (benzo[a]pyrene) hydroxylase (AHH) in the S9 fraction from Aroclor-treated rat livers and also inhibit(s) the mutagenic activity of benzo[a]pyrene (B(a)P) in the Ames test. The extract (fraction S30) was subjected to initial fractionation by thermal treatment, 3 24-h cycles of dialysis and ultrafiltration. The antigenotoxic activity of fraction S30 amounted to 98% and was unchanged by thermal treatment (100 degrees C, 10 min). Both the dialysate and the dialysis fluid inhibited the mutagenic effect of B(a)P by 48.4 and 48% respectively. The microsomal subfraction inhibited the mutagenicity only in 10%, and the postmicrosomal subfraction in 68%. It is concluded that the extract from wheat sprouts contains at least 2 heat-resistant compounds (or groups of compounds) located within the cell cytosol and showing antimutagenic activity: one group is of low molecular weight and another of high MW. Alternatively, low-molecular compounds could either be free or bound to high-molecular compound(s). PMID- 3050503 TI - Evaluation of the mutagenic activity of some phenanthroisoquinolines in Salmonella typhimurium. AB - The mutagenic activity of 3 aza-polycyclic aromatic hydrocarbons, which are suspected of being environmental pollutants, was assessed using the Salmonella assay. The compounds tested were phenanthro[9,10-g]isoquinoline, phenanthro[2,3 h]isoquinoline, and phenanthro[3,2-h]isoquinoline. None of the compound was mutagenic in the absence of S9, but all were mutagenic in the presence of S9. PMID- 3050504 TI - Evaluation of azo food dyes for mutagenicity and inhibition of mutagenicity by methods using Salmonella typhimurium. AB - The mutagenicity of 4 azo dyes (FD&C Yellow No. 5, FD&C Yellow No. 6, FD&C Red No. 40 and amaranth) that are widely used to color food has been evaluated. 4 different methods were used: (1) the standard Ames plate-incorporation assay performed directly on the dyes in the absence of S9 and in the presence of rat- or hamster-liver S9; (2) application of the standard plate assay to ether extracts of aqueous solutions of the dyes; (3) a variant of the standard assay, using hamster liver S9, preincubation, flavin mononucleotide (FMN) and other modifications designed to facilitate azo reduction; and (4) reduction of the dyes with sodium dithionite, followed by ether extraction and the standard plate assay. Assays that include chemical reduction (methods 3 and 4) were included because azo compounds ingested orally are reduced in the intestine with the release of free aromatic amines. No mutagenic activity was seen for any of the azo dyes tested by using the standard Ames plate assay (method 1). Ether extracts of some samples of FD&C Yellow No. 6, FD&C Red No. 40 and amaranth were active (method 2), but only at high doses, generally 250 mg-equivalents or more per plate. These results indicate the presence of low levels of ether-extractable mutagenic impurities. The FMN preincubation assay (method 3) gave negative results for all dye samples tested. Most batches of FD&C Red No. 40 tested had mutagenic activity that was detectable when the ether extract of less than 1 mg of dithionite-reduced dye was plated in the presence of S9 (method 4). This finding implies that an impurity in these samples of FD&C Red No. 40 can be reduced to yield an ether-extractable mutagen. Dithionite-reduced samples of FD&C Yellow No. 6 and amaranth showed ether-extractable mutagenic activity only at much higher doses than those at which activity was seen with most dithionite reduced samples of FD&C Red No. 40 (method 4). FD&C Yellow No. 5 showed no mutagenic activity with this method. Mutagenic activity was not detected when FD&C Red No. 40 was tested by using the azo reduction preincubation assay with FMN (method 3).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3050505 TI - Is dichlorvos a carcinogenic risk for humans? PMID- 3050506 TI - Verapamil as a co-mutagen in the Salmonella/mammalian microsome mutagenicity test. AB - Verapamil is a calcium-channel blocking agent, commonly used for chronic treatment of heart conditions. We have previously demonstrated that verapamil acts as a co-mutagen in a bacterial mutagenicity test for some experimental anilinoacridine antitumour drugs. Within the anilinoacridines series there are several compounds which are apparently non-mutagenic (or very weak mutagens) in the absence of verapamil, but strong mutagens in its presence. We have now tested a wider range of materials for verapamil enhancement of mutagenicity, to include some of those to which persons on verapamil therapy might be exposed through life style or occupation. Some verapamil enhancement of mutagenicity was seen with most mutagenic compounds including anticancer drugs, antiparasitic agents, one biological stain and one hair dye. A number of tricyclic antidepressants and biological stains were tested and found to be non-mutagenic. If these results extrapolate to mammalian cells, long-term verapamil therapy could potentially increase the effects of certain environmental mutagens. PMID- 3050507 TI - Relationships between structure of nitrated arenes and their mutagenicity in Salmonella typhimurium; 2- and 2,7-nitro substituted fluorene, phenanthrene and pyrene. AB - In order to elucidate the mechanisms of mutagenic activation of nitroarenes, we studied the relationships between the mutagenic potency and chemical structure of 2-nitro- and 2,7-dinitro-arenes including nitrated fluorene (Fl), dihydrophenanthrene (DHPh), phenanthrene (Ph), tetrahydropyrene (THPy), dihydropyrene (DHPy) and pyrene (Py) together with 9-NO2-Ph, 1-NO2-Py and 1.3 diNO2-Py. The mutagenicity tests were carried out on Salmonella typhimurium TA98, TA98NR and TA98/1,8-DNP6 in the absence of S9 mix. The order of mutability of mononitro- and dinitro-arenes in TA98 is as given below: 2-NO2-THPy less than 2 NO2-Fl less than 2-NO2-DHPh less than 9-NO2-Ph less than 2-NO2-Ph less than 2-NO2 DHPy less than 1-NO2-Py less than 2-NO2-Py, and 2,7-diNO2-DHPh less than 2,7 diNO2-Fl less than 2,7-diNO2-THPy less than 2,7-diNO2-Ph less than 2,7-diNO2-DHPy less than 2,7-diNO2-Py less than 1,3-diNO2-Py. 9-NO2-Ph and 1-NO2-Py, which have been detected in environmental samples, are not as potent mutagens as 2-nitrated phenanthrene and pyrene, respectively. 2-NO2THPy (37.7 rev/nmole) was a weak mutagen, but 2,7-diNO2-THPy (3197 rev/nmole) was as potent a mutagen as 2,7-diNO2 (3925 rev/nmole). Tetrahydropyrene has a twisted form in its structure. 1,3-diNO2 Py (99660 rev/nmole) was more mutagenic than 2,7-diNO2-Py (37960.0 rev/nmole), and their mutagenicities were correlated with the behavior of the K-band in their UV spectra by the introduction of nitro groups on pyrene. PMID- 3050508 TI - N-nitroso-butyl-3-carboxypropyl-amine (BCPN), a urinary bladder carcinogen, non mutagenic in S. typhimurium. PMID- 3050509 TI - AAEE minimonograph #29: automatic quantitative electromyography. AB - The present status of different computerized methods of automatic quantitative electromyography are reviewed. Interference pattern methods-turns analysis, spectral analysis-are efficient, but the results usually cannot be directly related to the physiological properties of the motor units. Integration analysis does not currently have a major role in diagnostic electromyography. Traditional measurement of single motor unit action potentials during weak contraction can be facilitated and made more objective with computer assistance, but only the lowest threshold motor units in the muscle are amenable to study. A new class of methodologies under development permit the decomposition of interference patterns into their constituent motor unit action potentials for measurement of configurational and behavioral properties. Patient data from these various methods can be statistically compared with normative data bases available on-line in computerized electromyographs. Both quantitative and quantitative electromyography have applications in the neuromuscular electrodiagnostic examination. PMID- 3050510 TI - Therapeutic trial of isaxonine in Duchenne muscular dystrophy. AB - A randomized double-blind therapeutic trial of isaxonine was completed over a 2 year period for 20 ambulant boys with Duchenne muscular dystrophy aged 5 1/2-10 years. The effect of the drug was monitored by measurement of walking times over 28 and 150 ft, motor ability score, MRC score based on 32 muscle groups, and myometry of 7 muscle groups. The drug had no significant effect on the progression of the disease. The trial had statistical power comparable to previous larger-scale multicenter trials. This reflected the low variability in the patients in relation to the magnitude of the overall deterioration. Measurements of muscle force (myometry and MRC score) had much greater statistical power than measurements of function (motor ability score and walking times) as analyzed by our methods. These observations have important implications for the design of future trials. PMID- 3050513 TI - [Stimulation of the filamentous growth of Candida albicans by aqueous humor]. PMID- 3050512 TI - AAEE minimonograph #30: electrophysiologic studies of myoclonus. AB - Definition as well as classification of myoclonus and electrophysiologic methods for investigating myoclonus were reviewed. Among the electrophysiologic techniques currently available in most laboratories, the EEG-EMG polygraph is the most essential one and can provide us with the most important information. Jerk locked averaging and evoked potential studies are useful for further investigating the pathophysiology of myoclonus and can be performed by using the same recording electrodes as those used for the polygraph. Jerk-locked evoked potentials and double-stimulation evoked potentials can be employed only for further investigating how cerebral cortex is involved in the generation of certain myoclonia. All these techniques can be used in proper combinations depending on the clinical features of the myoclonus in question, the purpose of the study, and the facilities available in each laboratory. These techniques also will be useful for following the clinical course during the treatment with antimyoclonus agents. PMID- 3050511 TI - Immunofluorescent evidence for presence of interleukin-1 in normal and diseased human skeletal muscle. PMID- 3050514 TI - The influence of some antifungal drugs on in vitro adherence of Candida albicans to human buccal epithelial cells. PMID- 3050515 TI - Properties and substrate specificity of a purine phosphoribosyltransferase from the human malaria parasite, Plasmodium falciparum. AB - The properties of a purine phosphoribosyltransferase from late trophozoites of the human malaria parasite, Plasmodium falciparum, are described. Enzyme activity with hypoxanthine, guanine and xanthine as substrates eluted in parallel during hydroxylapatite, size exclusion and DEAE-Sephadex chromatography as well as during chromatofocusing experiments. Furthermore, enzyme activity with all three purine substrates changed in parallel during heat inactivation of enzyme preparations and upon cold storage (4 degrees C) of the enzyme. When considered together, these results support the view that the phosphoribosyltransferase is capable of utilizing all three purine bases as substrates. Additional characterization revealed that the apparent molecular weight and isoelectric point of this enzyme are 55,500 and 6.2, respectively, and that the apparent Km for 5-phosphoribosyl-1-pyrophosphate ranges from 13.3 to 21.4 microM, depending on the purine base serving as substrate. The apparent Km values for hypoxanthine, guanine and xanthine were found to be 0.46, 0.30 and 29 microM, respectively. Other experiments showed that several divalent cations and sulfhydryl reagents produce a marked reduction of enzyme activity whereas dithiothreitol activates the enzyme. It should be noted that the ability to utilize xanthine as a substrate serves to distinguish the P. falciparum enzyme from its counterpart in the parasite's host cell, the human erythrocyte. The human enzyme shows only barely detectable activity with xanthine while the parasite enzyme displays similarly high levels of activity with all three purine substrates. Thus, the parasite enzyme might prove to be selectively susceptible to inhibition by xanthine analogs and related compounds. PMID- 3050516 TI - Blood glucose levels in Malawian children before and during the administration of intravenous quinine for severe falciparum malaria. AB - Hypoglycemia may develop in patients with severe untreated malaria and can complicate the course of treatment with parenteral quinine as a result of quinine induced hyperinsulinemia. Intravenous quinine is used increasingly as the therapy of choice in patients with severe malaria, most of whom are children. To assess the importance of both pretreatment and quinine-related hypoglycemia in children in an area in which the disease is endemic, we prospectively studied 95 Malawian children with falciparum malaria and altered consciousness who were treated with intravenous quinine. Nineteen patients had hypoglycemia before treatment. Seven (37 percent) died, and five of the survivors (26 percent) had neurologic sequelae. The corresponding values for patients who were initially normoglycemic were 4 percent and 4 percent, respectively (P less than 0.0001). Hypoglycemia was associated with low plasma insulin concentrations and with elevated plasma concentrations of lactate, alanine, and 5'-nucleotidase--a finding that suggests that impaired hepatic gluconeogenesis but not hyperinsulinemia contributes to the pathogenesis of pretreatment hypoglycemia. All patients were given quinine dihydrochloride in a 5 percent dextrose infusion, and those with hypoglycemia received 50 percent dextrose. Hypoglycemia recurred in seven of the patients with pretreatment hypoglycemia, but these episodes were also not associated with hyperinsulinemia. Of the 76 children who were initially normoglycemic, none became hypoglycemic during the course of treatment with intravenous quinine. We conclude that hypoglycemia is a frequent complication of falciparum malaria in children and that it reflects severe disease and is associated with a poor prognosis. We did not find it to be a complication of quinine treatment. PMID- 3050519 TI - Exchanging donor kidneys. PMID- 3050518 TI - Pathogenesis of sodium and water retention in high-output and low-output cardiac failure, nephrotic syndrome, cirrhosis, and pregnancy (1) PMID- 3050520 TI - Pulsatile secretion of insulin. PMID- 3050517 TI - Vancomycin, ticarcillin, and amikacin compared with ticarcillin-clavulanate and amikacin in the empirical treatment of febrile, neutropenic children with cancer. AB - We assessed two antibiotic regimens--vancomycin, ticarcillin, and amikacin, as compared with a vancomycin placebo, ticarcillin-clavulanate, and amikacin--as initial empirical therapy for febrile, neutropenic children with cancer. In a randomized, double-blind clinical trial, the planned 10-day treatment was unsuccessful in 15 percent of the vancomycin, ticarcillin, and amikacin group (n = 53), as compared with 38 percent of the group receiving ticarcillin-clavulanate and amikacin (n = 48) (P = 0.010). Of 10 episodes of breakthrough bacteremia, 9 (1 fatal) occurred in patients treated with ticarcillin-clavulanate and amikacin (P = 0.006). Each of the 10 microbial isolates was a gram-positive bacterium with similar susceptibilities to vancomycin and ticarcillin-clavulanate in vitro. Both regimens were well tolerated. None of the patients had detectable renal dysfunction, but those receiving vancomycin, ticarcillin, and amikacin were more likely to have twofold increases in serum hepatic-enzyme activity. Rashes consistent with the "red-man" syndrome occurred in three patients upon the infusion of vancomycin and in three others who received a placebo. We conclude that the combination of vancomycin, ticarcillin, and amikacin is more effective than ticarcillin-clavulanate and amikacin as empirical antibiotic therapy in clinical settings in which gram-positive bacteremias are a serious problem. PMID- 3050521 TI - Photodynamic therapy for bladder cancer: ethics of a collaborative trial in China. PMID- 3050522 TI - Aspirin, heparin, or both to treat acute unstable angina. AB - We tested the usefulness of aspirin (325 mg twice daily), heparin (1000 units per hour by intravenous infusion), and a combination of the two in the early management of acute unstable angina pectoris in a double-blind, randomized, placebo-controlled trial involving 479 patients. The patients entered the study as soon as possible after hospital admission (at a mean [+/- SD] of 7.9 +/- 8.0 hours after the last episode of pain), and the study was ended after 6 +/- 3 days, when definitive therapy had been selected. Major end points--refractory angina, myocardial infarction, and death--occurred in 23, 12, and 1.7 percent of the 118 patients receiving placebo, respectively. Heparin was associated with a decrease in the occurrence of refractory angina (P = 0.002). The incidence of myocardial infarction was significantly reduced in the groups receiving aspirin (3 percent; P = 0.01), heparin (0.8 percent; P less than 0.001), and aspirin plus heparin (1.6 percent, P = 0.003), and no deaths occurred in these groups. There were too few deaths overall to permit evaluation of the effect of treatment on this end point. The combination of aspirin and heparin had no greater protective effect than heparin alone but was associated with slightly more serious bleeding (3.3 vs. 1.7 percent). We conclude that in the acute phase of unstable angina, either aspirin or heparin treatment is associated with a reduced incidence of myocardial infarction, and there is a trend favoring heparin over aspirin. Heparin treatment is also associated with a reduced incidence of refractory angina. PMID- 3050523 TI - Pathogenesis of sodium and water retention in high-output and low-output cardiac failure, nephrotic syndrome, cirrhosis, and pregnancy (2) AB - This article has analyzed the pathogenesis of sodium and water retention in several circumstances. The initiator of retention has been proposed to be either a fall in cardiac output (e.g., low-output cardiac failure and vasoconstrictor hypovolemic nephrotic syndrome) or peripheral arterial vasodilatation (e.g., high output cardiac failure, cirrhosis, arteriovenous fistula, and pregnancy). In the only state discussed, in which the kidney is diseased and not merely responding to extrarenal reflexes--i.e., nephrotic syndrome--intrarenal mechanisms may predominate and lead to expansion of the arterial vascular tree and suppression of the renin-angiotensin-aldosterone system (i.e., hypervolemic nephrotic syndrome). Otherwise, when kidneys are healthy, either a fall in cardiac output or peripheral arterial vasodilatation may diminish arterial vascular filling and thereby initiate a series of hemodynamic and hormonal events that result in renal sodium and water retention (Fig. 7). Finally, the approach presented in this article should be considered to be a vantage point from which to evaluate states of sodium and water retention, but not to be an exclusive position. PMID- 3050525 TI - Variation in adhesion and cell surface hydrophobicity in Candida albicans white and opaque phenotypes. AB - A previous study had established that a select group of pathogenic isolates of Candida albicans was capable of switching heritably, reversibly and at a high frequency (10(-2) to 10(-3)) between two phenotypes ('white' or 'opaque') readily distinguishable by the size, shape, and color of colonies formed on agar at 25 degrees C. This paper describes experiments designed to determine the ability of these two phenotypes to attach to buccal epithelial cells (BECs) and plastic, and to compare the cell surface hydrophobicities of white and opaque phenotypes from three clinical isolates. 'White cells' were found to be significantly more adhesive to BECs, and a strong correlation was also found between phenotype adhesiveness and the percentage of BECs to which C. albicans had attached. The percentage of BECs with one or more attached C. albicans was approximately 90% for the white phenotype and approximately 50% for the opaque phenotype. 'Opaque cells', in contrast, were twice as hydrophobic as white cells, and the percentage of opaque cells bound to BECs by coadhesion was also double that of white cells. The differences in adhesion to plastic between the two phenotypes were not statistically significant and there was no distinct trend to suggest which phenotype might be more adhesive to plastic. These results indicate that several factors are involved in the adhesion of C. albicans to plastic, and confirm the hypothesis that cell surface hydrophobicity is of minor importance in direct adhesion to epithelial cells but that it may contribute to indirect attachment to epithelial cells by promoting yeast coadhesion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3050526 TI - Detection of Candida sp. mannan antigen by indirect ELISA-inhibition. In vitro crossed reactivity among mannans obtained from C. albicans A, C. albicans B, and C. tropicalis. AB - We have obtained mannans from four Candida species: C. albicans A, C. albicans B and C. tropicalis; anti-mannan sera against C. albicans A, C. albicans B and C. tropicalis were obtained by immunizing rabbits subcutaneously with the respective yeast extract. The efficacy of these sera in reacting with mannans obtained from three Candida sp. has been proven by indirect ELISA-inhibition. Any of three immune sera can be used to detect mannan antigen from the three Candida sp. tested. This confirms the existence of crossed reactivity and the possibility of detecting mannan antigen in serum from patients infected by different Candida sp., although we had only one immune serum and one Candida mannan. PMID- 3050524 TI - Immunization of pregnant women with a polysaccharide vaccine of group B streptococcus. AB - Immunization of pregnant women with a polysaccharide vaccine of group B streptococcus is a promising strategy for the prevention of perinatal infections caused by group B streptococci. To explore the feasibility of this strategy, we vaccinated 40 pregnant women at a mean gestation of 31 weeks with a single 50 microgram dose of the Type III capsular polysaccharide of group B streptococcus. The only adverse effect detected was a mild local reaction in nine women (22 percent). Of the 35 women with low or unprotective antibody levels before immunization (less than 2 micrograms per milliliter), 20 (57 percent) responded to the vaccine. The geometric mean antibody level rose from 1.3 to 7.1 micrograms per milliliter four weeks after vaccination (P less than 0.02), and these levels persisted at delivery and three months post partum. Sixty-two percent of the vaccine-induced immunoglobulin in the mothers was IgG, which readily crosses the placenta. Infant antibody levels in cord serum correlated directly with maternal antibody levels at delivery (r = 0.913, P less than 0.001). Of the 25 infants born to women who responded to the vaccine, 80 percent continued to have protective levels of antibody at one month of age and 64 percent had protective levels at three months. Serum samples from infants with greater than or equal to 2 micrograms of antibody to Type III group B streptococcus per milliliter uniformly promoted efficient opsonization, phagocytosis, and bacterial killing in vitro of Type III strains. This effect could be mediated exclusively by the alternative complement pathway. Although this vaccine with an overall response rate of 63 percent is not optimally immunogenic, we conclude that maternal immunization is feasible and can provide passive immunity against systemic infection with Type III group B streptococcus in the majority of newborns. Larger trials with better vaccines will be required to evaluate the safety and clinical effectiveness of this strategy. PMID- 3050527 TI - Experimental aspergillosis in Japanese quails (Coturnix coturnix japonica). Clinical signs and haematological changes. AB - Intratracheal inoculation of 2-week-old quail chicks with Aspergillus fumigatus resulted in the development of clinical signs within 24 h of infection. These were characterized by anorexia, depression, accelerated respiration and gasping followed by death. The acute course of the disease lasted for 7-10 days followed by recovery in the surviving chicks. The overall mortality during a 6-week observation period was 20%. Although the mean body weight of A. fumigatus infected quail chicks continued to be slightly lower throughout the experiment but the difference, in comparison to controls, was not significant except at 42 days post-infection. There was no appreciable difference in the mean values of Hb, TEC, PCV, MCV, MCH and MCHC between the infected and control chicks at any stage of infection but TLC revealed a leucocytosis from 2-7 days which was the result of increase in the relative percentage of heterophils and decrease in lymphocytes. PMID- 3050528 TI - Simple method for separating the chlamydoconidia of Candida albicans from its mycelium. AB - Different physical and chemical methods were used to detach the chlamydoconidia of Candida albicans from its mycelium. The action of concentrated H2SO4 acid for a 4-min period on cultures lysed both the mycelium and the outer but not the inner wall layer of the chlamydoconidia. The sulfuric acid procedure is recommended as the best method to obtain mycelium free chlamydoconidia because of its simplicity, rapidity and low cost. PMID- 3050529 TI - Existence of syphilis in a Pleistocene bear. PMID- 3050530 TI - Pancreatic amylin and calcitonin gene-related peptide cause resistance to insulin in skeletal muscle in vitro. AB - Insulin resistance occurs in a variety of conditions, including diabetes, obesity and essential hypertension, but its underlying molecular mechanisms are unclear. In type 2 (non-insulin-dependent) diabetes mellitus, it is insulin-resistance in skeletal muscle, the chief site of insulin-mediated glucose disposal in humans, that predominantly accounts for the low rates of glucose clearance from the blood, and hence for impaired glucose tolerance. Human type 2 diabetes is characterized by a decrease in non-oxidative glucose storage (muscle glycogen synthesis), and by the deposition of amyloid in the islets of Langerhans. Amylin is a 37-amino-acid peptide which is a major component of islet amyloid and has structural similarity to human calcitonin gene-related peptide-2 (CGRP-2; ref. 8). CGRP is a neuropeptide which may be involved in motor activity in skeletal muscle. We now report that human pancreatic amylin and rat CGRP-1 are potent inhibitors of both basal and insulin-stimulated rates of glycogen synthesis in stripped rat soleus muscle in vitro. These results may provide a basis for a new understanding of the molecular mechanisms that cause insulin resistance in skeletal muscle. PMID- 3050532 TI - National Institutes of Health Consensus Development Conference on the Management of Clinically Localized Prostate Cancer. Bethesda, Maryland, June 15-17, 1987. PMID- 3050531 TI - How eukaryotic transcriptional activators work. AB - A specific protein, bound to DNA, can activate transcription of a wide array of genes in many eukaryotes. Further analysis suggests a general outline for how eukaryotic transcriptional activators function and are controlled. PMID- 3050533 TI - Randomized series of treatment with surgery versus radiation for prostate adenocarcinoma. AB - In the early 1970s, a multicentered cooperative group effort was established by urologists and oncologists to examine the relative disease control provided by surgery, radiation therapy, or observation for patients with localized or regional disease. The data derived from this trial were controversial because they 1) did not support previous concepts regarding the relative impact of treatment and 2) raised provocative questions as to the interpretation of previous institutional reports that promoted a single treatment modality. The data from the randomized trial demonstrated that: 1) bipedal lymphangiography could not demonstrate accurately the presence or absence of microscopic involvement of pelvic lymphatic structures, 2) treatment selection should be based on the anatomic distribution of disease; 3) a clinician's use of first appearance of local or distant disease in a patient who was supposedly disease free after receiving the chosen therapy served as an accurate way to define the impact of the initial treatment; 4) radical surgery was more effective than radiation therapy in controlling disease that was clinically confined to the primary organ of origin; and 5) the apparent disease control produced by radiation on large-volume, localized disease might only reflect the natural history of the disease. PMID- 3050534 TI - Chemotherapy for prostate carcinoma. AB - We have evaluated the role of chemotherapy for the treatment of prostate carcinoma. The data of patients with endocrine-resistant stage D2 disease indicate that clinical benefits in such patients are at best marginal. Despite the controversies involved in the assessment of response in this disease, in this review we show that in over 3,000 patients eligible for evaluation, less than 10% had complete or partial responses to various treatment regimens. Survival evaluation on all prospective randomized clinical trials showed no advantages in favor of any treatment tested and, moreover, in 2 of such studies involving various single agents, survival was not better than a "no chemotherapy" control arm. Because of these data, we conclude that chemotherapy is not indicated as an adjuvant treatment for patients with localized prostate cancer. Although patients with prostate cancer frequently respond to androgen deprivation procedures, preclinical and clinical data strongly suggest the existence of endocrine independent cell clones, which supports further testing with nonhormonal cytotoxic treatment. A close multidisciplinary interaction is a prerequisite for development of new effective systemic treatment in this disease. PMID- 3050535 TI - Hormone therapy for prostate cancer: results of the Veterans Administration Cooperative Urological Research Group studies. AB - Between 1960 and 1975, the Veterans Administration Cooperative Urological Research Group conducted a consecutive series of 3 major randomized clinical trials comparing various endocrine treatments for newly diagnosed prostate cancer patients. Six major conclusions concerning hormonal treatment emerged from these studies: 1) increased hazard of cardiovascular death after therapy with 5 mg diethylstilbestrol (DES); 2) orchiectomy plus DES no better than orchiectomy or DES alone; 3) equivalent effect of 1.0 and 5.0 mg DES on cancer; 4) reduced cardiovascular hazard from therapy with 1.0 mg DES; 5) Premarin and Provera no better than 1.0 mg DES at doses studied; 6) decisions about hormone treatment at diagnosis dependent on patient characteristics, mainly age and Gleason grade. In this paper, these studies are reviewed briefly and data are presented to support these conclusions. Some tentative treatment recommendations are proposed. PMID- 3050536 TI - Hormonal therapy for locally advanced prostate cancer. AB - A patient with locally advanced prostate cancer (stages C and D1) has a poor prognosis with a high risk of developing and dying of distant metastases. Hormonal therapy is the major form of systemic therapy for metastatic (stage D2) prostate cancer. The most commonly used forms of hormonal therapy are orchiectomy, diethylstilbestrol, and luteinizing hormone releasing hormone, agonists that prevent the stimulation of tumor cells by testosterone. They produce a 60%-80% symptomatic or objective response rate, but their ability to prolong overall survival remains uncertain. Surgical adrenalectomy, hypophysectomy, and pharmacologic adrenal suppression prevent the clinically less significant adrenal androgen stimulation of tumor cells. Antiandrogens competitively inhibit the interaction between androgens and cytosolic androgen receptors. Complete androgen blockade (luteinizing hormone releasing hormone agonist and antiandrogen) was initially espoused to be superior to single-agent hormonal therapy, but preliminary results from a multigroup randomized trial suggest that it has only a minimal advantage. The benefit of hormonal therapy in stages C and D1 prostate cancer at the time of diagnosis has not been clearly established. Available studies are few, and most often they are uncontrolled or include only small numbers of patients. However, they suggest that the early use of hormonal therapy prolongs disease-free survival but does not prevent ultimate disease progression or prolong overall survival. Hormone receptor assays may be helpful in the selection of patients who would benefit from early hormonal therapy. PMID- 3050537 TI - Fine-needle aspiration of the prostate. AB - Fine-needle aspiration biopsy of the prostate has been used for over 20 years as the prostate biopsy technique of choice throughout much of Europe. However, this technique has only recently gained acceptance in the United States. There is now an enlarging body of data confirming that fine-needle aspiration can be viewed as a safe, accurate, and reliable means for detection and diagnosis of prostate cancer. This review summarizes the European and United States experience with fine-needle aspiration and supports the utility of this biopsy technique as an important addition to the urologist's diagnostic armamentarium. PMID- 3050539 TI - Consensus statement: the Management of Clinically Localized Prostate Cancer. National Institutes of Health Consensus Development Panel. PMID- 3050538 TI - Application of flow cytometry and automated image analysis to the study of prostate cancer. AB - Flow cytometry and image analysis are complementary quantitative cytologic techniques that have demonstrated utility in the assessment and analysis of prostate cancer and other urologic and nonurologic tumors. This review is intended to assess the current state of the art and to project future directions and applications of these modalities in the pathologic assessment of prostate cancer. Special attention is directed toward the topics of prostate cancer detection and diagnosis, determination of patient prognosis, and the monitoring of patient response to therapy. We recommend that 1) flow cytometry and image analysis be used to determine pathologic parameters that will help in predicting poor patient prognosis and 2) quantitative cytologic determinations of DNA content be included in clinical trials so that their ultimate role in monitoring patient response to therapy can be determined. This knowledge will allow the development of protocols designed to test the value of earlier institution of multimodal therapy in high-risk populations. PMID- 3050540 TI - Noninvasive imaging for staging of prostate cancer: magnetic resonance imaging, computed tomography, and ultrasound. AB - The diagnosis of prostate carcinoma by imaging is still fraught with problems, even with the advent of highly sophisticated techniques. Despite enthusiastic preliminary reports, no one imaging method reliably screens for this condition. The staging of prostate carcinoma is feasible, but the best imaging method remains a subject of debate. The transabdominal sonographic approach lacks the resolution required for detailed intraglandular anatomic delineation. The transrectal sonographic approach excels in guiding needle biopsy and in evaluating transcapsular and seminal vesicle extension of known tumors. Computed tomography lags behind other tomographic imaging modalities in its accuracy for local tumor staging, but it is excellent, although nonspecific, in the detection of lymph node enlargement. Magnetic resonance detects abnormalities in the prostate in a high percentage of cases but is nonspecific. However, it can stage prostate carcinoma and detect lymphadenopathy reliably. PMID- 3050541 TI - Consensus Development Conference on the Management of Clinically Localized Prostate Cancer. Overview: historical and contemporary. AB - Recognition of the clinical importance of prostate cancer undoubtedly was delayed by the failure of clinicians or pathologists to distinguish consistently between benign and malignant prostatic growths until well into the 19th century. White used castration for prostatic enlargements in 1895, but Huggins and Hodges first placed endocrine therapy on a rational basis in 1941. Although a number of surgeons had attempted excision of prostate cancer, Young is credited with planning and performing the first radical perineal prostatectomy in 1904. Orthovoltage irradiation and various techniques of interstitial and intracavitary radium therapy were used in the treatment of prostate cancer early in the 20th century, but it was the development of megavoltage irradiation that reopened the door to the exploration of irradiation for localized prostate cancer following World War II. Endocrine manipulation, surgery, and irradiation remain the keystones of treatment. The management of prostate cancer is controversial for several reasons: 1) The disease occurs in an age range in which competing causes of mortality are high. 2) The natural evolution of the disease is varied, often long, and not consistently predictable. 3) Long-term survival has been reported for each of the principal modes of therapy, but randomized controlled studies have been limited. Uniformity in histologic grading, clinical staging, and evaluation of response to treatment would improve the quality of the data. Predictions of host life expectancy, tumor growth rate, metastatic potential, and tumor responsiveness to irradiation and endocrine therapy would enhance the rationale of treatment. PMID- 3050542 TI - External-beam radiation therapy for clinically localized prostate cancer: patterns of care studies in the United States. AB - Data are presented from the Patterns of Care Study and other sources that define the role of external-beam irradiation in the management of localized prostate cancer as practiced in the United States as a whole. Patients must be treated with complex treatment techniques and high-energy linear accelerators and careful adjustment of radiation dose. Transurethral resection of the prostate should be avoided in the intermediate and poorly differentiated subgroup of stage C patients. The excellent 5- and 10-year survival for patients treated by radiation therapy is demonstrated for all stages of prostate cancer and for T1 or early stage B patients. It is noted that the national averages for survival have improved between 1973 and 1978. Stages A2 and B patients with negative lymph node dissections show freedom from recurrence that is equal to patient reports for radical surgery. Complications resulting from radiation therapy were modest, and potency was maintained in 73% of the patients. Adjuvant irradiation is necessary for pathologic stage C patients after recovery from surgery. Radiation therapy is equally effective though less costly than surgery for early prostate cancer. A particular need of future research is the study of the patterns of care in the United States regarding the surgical management of prostate cancer so that health professionals can determine if this care is generally available throughout the United States and if good outcome and acceptable morbidity result after it is given. PMID- 3050544 TI - Roster. North Carolina Medical Society 1988-1989. PMID- 3050543 TI - Local control of prostate cancer with radiotherapy: frequency and prognostic significance of positive results of postirradiation prostate biopsy. AB - The best available data indicate that, although it is imperfect, the postirradiation biopsy performed at a sufficient interval after radiotherapy can provide accurate prognostic information useful in the determination of the success or failure of radiotherapy in an individual patient as well as the measurement of overall efficacy of any particular radiotherapeutic regimen. Needle biopsy of the prostate was performed routinely in 510 patients with clinical stage A2, B, or C1 prostate cancer treated with a combination of radioactive gold seed implantation and external-beam irradiation. Of the 140 patients who had one or more needle biopsies performed 6-36 months after completion of radiotherapy, who had no evidence of local recurrence or distant metastases at the time of biopsy, and who had received no hormonal therapy before documented recurrence of the tumor, 45 (32%) had one or more biopsies positive for cancer. The frequency of positive biopsy results correlated significantly with the size of the local tumor but not with the grade. The correlation between biopsy results and the eventual development of recurrence was highly significant. If any biopsy was positive, 60% of the patients eventually developed local recurrence; if all biopsies were negative, only 19% developed local recurrence during the period of follow-up. The poor prognosis associated with a positive biopsy result was found within almost every subset of stage, grade, or nodal status examined although the results varied because of the small number of patients in some groups. PMID- 3050545 TI - [Persistent extrauterine pregnancy]. PMID- 3050546 TI - [The regulation of potassium metabolism]. PMID- 3050548 TI - [Pregnancy and antiepileptic agents; the value of prenatal ultrasonography studies for the detection of congenital malformations]. PMID- 3050547 TI - [Potassium supplementation in diuretics]. PMID- 3050549 TI - [Drug treatment of the postmenopause]. PMID- 3050550 TI - [Consensus diagnosis of atopic syndrome]. PMID- 3050551 TI - [Malignant histiocytosis, true histiocytic lymphoma, acute monoblastic leukemia; forms of degeneration in the monocytoid series]. PMID- 3050552 TI - [Suitability in health services: a million-dollar question]. PMID- 3050553 TI - [Lung embolism in diagnostic perspective]. PMID- 3050555 TI - [Decrease in human immunodeficiency virus antigen levels in the cerebrospinal fluid during zidovudine treatment in patients with AIDS]. PMID- 3050556 TI - [Dialysis and kidney transplantation in children]. PMID- 3050554 TI - [The artificial bringing together of female and male gametes in the fallopian tubes (the GIFT method): a new treatment for infertility]. PMID- 3050558 TI - [Antibiotics policy in acute tonsillitis managed by the family practitioner; a decision analysis]. PMID- 3050557 TI - [Nephrotic syndrome in the first year of life]. PMID- 3050559 TI - Hyperbaric oxygen and multiple sclerosis--a review. PMID- 3050560 TI - The Healing Arts. By Czar Johnson. Nebraska Medical Journal, January 1937. PMID- 3050561 TI - [Projections of fields 5 and 7 in the subdivision of the sensorimotor area of the cat cerebral cortex]. AB - Projections of the parietal cortex (area 5 and 7) to the subdivisions of the sensorimotor cortex have been studied with axonal degeneration method. It was shown that density of distribution of the association fibres is different in the areas of the parietal and sensorimotor cortex. Area 5 projects mainly to the posterolateral part of the cruciate sulcus (areas 4fu, 4 delta); its projections to areas 4y, 4sfu and 6iffu, 6aa, 6ab are less pronounced. Area 7 is connected mostly with the medial part of the lower lip of the cruciate sulcus (areas 6iffu, 6aa, 6ab) and its projection to areas 4fu and 4 delta is less pronounced. Poor interrelation between area 5 and areas SI (2, 3a, 3b) and no projection to SII (second somatosensory area of Wolsey) were observed. No projections were found from area 7 to areas SI and SII. PMID- 3050562 TI - [A. A. Ukhtomskii and I. P. Pavlov]. PMID- 3050563 TI - [Modern approaches to the study of cellular and synaptic mechanisms of the so called dominant]. PMID- 3050565 TI - [Hemispheric functions and psychiatric diseases]. PMID- 3050564 TI - [A. A. Ukhtomskii and theoretical physiology]. PMID- 3050566 TI - [Empirical research of the classification of psychiatric disorders]. PMID- 3050568 TI - ["Desactualization weakness" of schizophrenic patients and its relation to productive-psychotic symptoms]. PMID- 3050567 TI - [Development of a systems theory of productive psychoses]. PMID- 3050569 TI - [Constituent differences of delusions in various psychotic illness states]. PMID- 3050570 TI - [Long-term effects of a combination of transdermal nicotine administration with behavior therapy for smoking cessation]. AB - Effects of a transdermal nicotine substitution on psychological smoking cessation were investigated in a double-blind prospective study. 131 smokers have been randomly assigned to three treatment conditions: All smokers underwent nine weeks of self-controlled smoking cessation. During 6 weeks one group was additionally treated with nicotine patches continuously releasing nicotine through the skin into the blood circuit. The second group received placebo patches; while the third group was treated with behavioral training alone. Treatment effects were measured by daily cigarette consumption. Follow-up investigations were performed 3, 6 and 12 months after therapy. Nicotine-treated subjects reached significantly higher abstinence rates during and at the end of treatment as well as during the follow-up period, than both placebo- and control-subjects. No severe side effects of plasters have been reported. The results thus indicate good therapeutic effectiveness of transdermal nicotine substitution. PMID- 3050571 TI - Alport's syndrome: risk of glomerulonephritis induced by anti-glomerular-basement membrane antibody after renal transplantation. AB - Two patients with Alport's syndrome developed glomerulonephritis induced by anti glomerular-basement-membrane (anti-GBM) antibody after renal transplantation. One patient presented at 5 months after transplantation the second patient 18 months after transplantation. Both grafts failed and the patients returned to dialysis. Our observations suggest that the patients with Alport's syndrome are at risk of developing anti-GBM antibody type glomerulonephritis in the transplanted kidney. This does not occur in the early period after transplantation, probably because of heavy immunosuppression. PMID- 3050572 TI - Paracetamol: a cause for analgesic nephropathy and end-stage renal disease. AB - 180 patients with end-stage renal disease (ESRD) on maintenance dialysis and those who had undergone renal transplantation were questioned retrospectively. 14 patients had consumed excessive quantities of analgesics (greater than 1 kg) prior to the institution of long-term dialysis or transplantation. Sonographic examination done on these patients indicated that 7 had renal papillary necrosis (RPN). The sonographic features were renal papillary calcifications surrounding the central sinus in a complete or incomplete garland pattern. In 5 of these patients RPN is attibutable to the excessive consumption of paracetamol. We have earlier reported 10 cases of RPN due to excessive consumption of paracetamol. Thus 15 cases of RPN attributable to paracetamol consumption (1.0-15.3 kg over a period ranging from 3 to 23 years) have been documented. It is concluded that paracetamol may assume an increasingly important role in the causation of analgesic nephropathy (AN) and ESRD. PMID- 3050573 TI - The retinoid status of kidney transplant recipients. PMID- 3050574 TI - Spontaneous streptococcal peritonitis in renal transplant recipient. PMID- 3050576 TI - CAPD, protective against developing dialysis-associated amyloid? PMID- 3050578 TI - [Involvement of the renal interstitium in systemic diseases (I)]. AB - Interstitial lesions which are frequent in systemic diseases such as systemic lupus erythematosus, Sjogren's syndrome, sarcoidosis and Dobrin's syndrome (acute interstitial nephritis with uveitis) are described as well as their clinical expression. The immune reactions which are recognized to be responsible for these lesions are discussed in respect to each of these diseases. PMID- 3050577 TI - Systemic lupus erythematosus and complement. AB - SLE is characterized by a host of immune abnormalities. It is not clear to date which of these are primary and which are secondary. The observation of a number of genetic defects suggests that they are primary. Multiple genetic defects may then lead to abnormal immune responses to common pathogens, antigens, or even autoantigens. As a result of this abnormal immune response, immune complexes form with resultant complement fixation and activation. These immune complexes interacting with cells and complement initiate an inflammatory response. One can also speculate that this inflammatory response represents a normal response to an abnormal event, or is also abnormal in the SLE patient. The ultimate result is tissue inflammation and often damage. While at present our therapy is aimed at controlling these secondary inflammatory phenomena mediated by immune complexes and complement, ultimately therapy may be more successful after the primary defects are corrected. PMID- 3050579 TI - [Functional renal effects of cyclosporin. Physiopathological mechanisms]. AB - Animals and humans undergoing a treatment with cyclosporin (CsA) show a reversible increase in renal vascular resistance and decrease in glomerular filtration rate. The causes of these abnormalities have not yet been established. In animals potential mechanisms for CsA induced renal functional impairment are an increase in urinary thromboxane A2 excretion, plasma renin activity and renal sympathetic nervous system activity and an enhancement of vasopressin stimulated Ca++ mobilisation and cell contraction in vascular smooth muscle cells. In human, the problem is far less clear. CsA induces an inhibition in PRA and urinary prostaglandins excretion. Furthermore CsA does not modify urinary and plasma levels of catecholamines. Whatever the mechanism underlying the vasoconstriction induced by CsA, the inhibition of PGI2 synthesis and angiotensin II formation may participate in the decrease in renal blood flow and glomerular filtration rate which is observed in patients receiving CsA. PMID- 3050575 TI - Sepsis and death after embolization of host kidneys in a resistant renal hypertension transplanted patient. PMID- 3050580 TI - [The captopril test in nonselected hypertensive patients: absence of value for screening of renovascular patients and for unilateral small kidneys]. AB - The decrease of blood pressure and the increase of plasma renin activity (PRA) after oral administration of captopril was evaluated in 104 consecutive hypertensive out-patients in whom the morphology of the renal arteries and parenchyma was assessed thanks to an intravenous digitalized angiography. Twenty five of these patients were excluded because of a natriuresis less than 50 mmol/24 h or non discontinuation of their treatment; 50 of these patients were classified as essential hypertension, 12 had renovascular disease (10 unilateral stenosis 2 of which significant; 2 bilateral significant stenosis); 9 had unilateral small kidneys (4 significant). The significance was ascertained on the PRA in the renal veins and/or the decrease of hypertension after surgery. The decrease of blood pressure after captopril was not different between the various groups. The increase after captopril of PRA was higher in unilateral significant lesions. However the highest post captopril PRA value was found in the essential hypertension group so that renovascular diseases could not be screened by higher post captopril PRA values. However taking into account that these latter were decreasing with age, a better discrimination of significant unilateral disease was possible in the hypertensive patients above 40 years of age. Furthermore once the diagnosis of unilateral kidney disease is established by radiological investigation, the captopril test allows to predict unilateral hypersecretion of renin with a sensitivity of 100% and a specificity of 78%. IN CONCLUSION: the captopril test does not allow to screen the non selected hypertensive patients for an efficient radiological investigation by intravenous digitalized angiography but may help to select the patients with unilateral renal disease for the renal venous PRA evaluation. PMID- 3050582 TI - [The cell biological characteristics of brain tumors]. PMID- 3050581 TI - [Multifocal osteoarthropathy associated with beta-2 microglobulin amylosis deposit in a patient with renal insufficiency treated exclusively with hemofiltration]. PMID- 3050583 TI - [Cell kinetics of human meningiomas and neurinomas--Ki-67 PAP stain]. AB - Seventeen surgically resected meningiomas and six neurinomas including one neurofibroma were labelled with frozen sections using a monoclonal antibody Ki 67, which reacts with a nuclear antigen expressed by proliferating cells. In thirteen classic non-malignant meningiomas, the percentage of stained cells ranged from 0.2 to 3.2% (mean, 1.4%). Those of three recurrencies consisted of anaplastic one and another one with anaplastic component, and one hemangiopericytic type ranged from 2.9 to 8.7% (mean, 4.8%). Four classic non malignant neurinomas and one neurofibroma had the range from 0.3 to 2.7% (mean, 1.3%). However, one anaplastic neurinoma showed the largest fraction (13.0%) of positive nuclei in all these tumors. These results show, in conclusion, that the determination of Ki-67 immunoreactivity in the cell nuclei of meningiomas and neurinomas can contribute to more objectiveness in histological grading and the higher positive rate could be one of various factors relating to recurrence. PMID- 3050584 TI - Transplanted gonadotropin-releasing hormone neurons promote pulsatile luteinizing hormone secretion in congenitally hypogonadal (hpg) male mice. AB - Congenitally hypogonadal (hpg) male mice are unable to synthesize biologically active gonadotropin-releasing hormone (GnRH). Implantation of normal fetal preoptic area tissue containing GnRH neurons into the third ventricle of adult hpg males significantly elevates pituitary levels of luteinizing hormone (LH) and corrects their hypogonadism. In all responding animals, immunoreactive GnRH neurons within the transplant innervate the median eminence of the host. To assess whether gonadal recovery in hpg hosts results from pulsatile secretion of GnRH from grafted neurons, we compared the pattern of variation in plasma LH levels in 19 hpg graft recipients with testicular growth to that of 10 normal adult mice. All animals were castrated prior to receiving an indwelling catheter in the jugular vein. Sequential blood samples were collected (t = 10 min) and assayed for LH. Pulsatile LH secretion was seen in 11 of 19 hpg hosts and in all control mice. While there was great variability between individual animals, measures of baseline LH, LH pulse amplitude and duration, interpulse interval, and LH pulse frequency revealed no difference between hpg graft recipients and normal castrates in their LH pulse pattern. Immunocytochemical analysis of the brain in hpg hosts suggested no correlation between any parameter of pulse activity and individual differences in GnRH cell number or GnRH fiber outgrowth into the median eminence. Sources of variation in LH secretion among graft recipients, and between hpg hosts and normal mice, are discussed. We suggest that transplanted GnRH neurons are capable of integration into a GnRH 'pulse generator' which can support a near-normal pattern of pulsatile LH secretion, leading to testicular growth and steroid production. PMID- 3050586 TI - Cerebral giant serpentine aneurysm: case report and review of the literature. AB - We present a case of cerebral giant serpentine aneurysm (GSA) and propose a definition of GSA. Our literature review disclosed only 16 cases, including our own, that fit our criteria. GSAs belong to the subgroup of giant aneurysms, but are distinct from giant saccular and fusiform aneurysms. We discuss their specific characteristics. PMID- 3050585 TI - Delayed sexual maturation induced by daily melatonin administration eliminates the LH response to naloxone despite normal responsiveness to GnRH in juvenile male rats. AB - Daily administration of melatonin (MT) markedly delays sexual maturation in the male Wistar rat. In this study, we have evaluated pituitary responsiveness to GnRH and the level of tonic inhibition by endogenous opioids in normal juvenile male rats and in rats with delayed sexual development induced by daily afternoon MT injection (100 micrograms, s.c.) starting at 20 days of life. Plasma LH responses to repetitive intravenous GnRH administration (100 ng/100 g body weight), or to different doses of GnRH administered subcutaneously (5-100 ng/100 g body weight) were normal in MT-treated rats both at 30 and 40 days of life despite significantly lower number of pituitary GnRH receptors and decreased pituitary gonadotropin content. One naloxone (NAL) injection (2.5-5.0 mg/kg, s.c.) produced a significant increase of plasma LH in normal 40- and 55-day-old rats, which was not seen in MT-treated rats of the same age. In contrast, no increase of plasma LH was seen in 30-day-old control rats nor in MT-treated rats at this age. Pretreatment with morphine sulfate (10 mg/kg, s.c.), or with the potent Met-enkephalin analog FK 33-824 (1.0 mg/kg, s.c.) prevented the NAL induced rise of plasma LH in control rats at day 40 of life. In all instances, plasma PRL levels were decreased after NAL both in untreated and in MT-treated rats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3050588 TI - Changes in the lateral ventricle with the head position: ultrasonographic observation. AB - Ultrasonographic examination was performed in 13 preterm, 10 normal term and 3 term brain-damaged infants to evaluate changes of the ventricular width with the head position. The lower side of the frontal horn was narrowed and the upper side dilated in all preterm and brain-damaged infants. These changes of the ventricular width may be due to gravity and soft brain. Therefore, it is very important to consider the transformation of the lateral ventricles with the head position when asymmetry of the lateral ventricle is identified in infants showing prematurity or brain damage. PMID- 3050589 TI - Excitatory amino acid neurotoxicity in the hippocampal slice preparation. AB - The effect of sustained activation of excitatory amino acid receptors on neuronal survival was studied using slices of adult rat hippocampus and light and electron microscopy. Kainate, N-methyl-D-aspartate, quisqualate, and ibotenate all produce signs of severe neurotoxicity within 90 min. Neuronal damage occurs in the form of perikaryal and dendritic swelling, cytoplasmic and nucleoplasmic disintegration, and plasma and nuclear membrane ruffling and collapse. The toxicity is restricted to intrinsic neuronal somata, dendrites and spines, while afferent axons, boutons and glia are spared. Although damage is generally distributed throughout all areas of hippocampus, kainate has little effect on pyramidal neurons in the CA2 region. Quantitative analysis of neuronal survival indicates that agonists induce dose-dependent damage over concentration ranges known to be excitatory. Based on selective antagonism by DL aminophosphonoheptanoate and the patterns of damage produced by each, N-methyl-D aspartate, kainate, and quisqualate trigger neurotoxicity by acting on distinct receptor classes. It is concluded that, in hippocampal slices, excitatory amino acids induce neurotoxicity in a similar manner to their actions in vivo. The results support the hypothesis that hippocampal neurotoxicity is initiated by excessive excitation, and provide another example of the capacity of adult hippocampal neurons for rapid structural modification. PMID- 3050590 TI - [Computerized ultrasonic angiography. Current applications and comparison with other noninvasive and invasive methods]. PMID- 3050591 TI - [Surgical prospects in dyskinesias of the pelvic floor and anal canal]. PMID- 3050587 TI - Magnetic resonance imaging in the evaluation of treatment in multiple sclerosis. AB - Magnetic resonance scans of 74 patients with multiple sclerosis participating in a controlled trial were compared 6 months before and at the end of a 24-32 months treatment period with either Cyclosporin A (n = 31) or Azathioprine (n = 43). Both qualitative rating and computation of lesion volume showed deterioration in more than 40% of the patients, while by clinical criteria only 10-30% were worse. No significant difference was noted when the two treatment groups were compared. If careful repositioning and standardized image parameters are used, MRI is an indispensable tool for the objective determination of disease progression in MS although it cannot replace clinical examination. PMID- 3050593 TI - [Essential sclerosing cholangitis. Description of a clinical case]. PMID- 3050592 TI - [Complications of mechanical sutures in colorectal surgery. 5-year experience]. PMID- 3050594 TI - [Reconstruction of the sphincter in a case of severe anal incontinence caused by perianal abscess]. PMID- 3050595 TI - [Treatment of perforated duodenal ulcer]. PMID- 3050596 TI - [Functional exclusion of the exocrine pancreas in pancreatic transplants: use of polymers in animal experiments]. PMID- 3050597 TI - [Role of sclerotherapy in bleeding esophageal varices in children with portal hypertension]. PMID- 3050598 TI - [Diagnosis and surgical therapy of pancreatic abscess]. PMID- 3050599 TI - [Computerized program for rapid evaluation of the critical surgical patient with multiple trauma]. PMID- 3050600 TI - [Anatomo-clinical considerations on lymphatic metastasis of cancer of the stomach]. PMID- 3050601 TI - [A case of cystic lymphangioma of the retroperitoneum]. PMID- 3050602 TI - [A rare case of carcinoid tumor of the gallbladder]. PMID- 3050603 TI - [Comparative study of mechanical and microsurgical anastomosis of small-diameter arteries]. PMID- 3050605 TI - [Technical details of closed submucosal hemorrhoidectomy]. PMID- 3050606 TI - [Solitary ulcer of the perforated sigmoid-rectum. Clinical contribution]. PMID- 3050604 TI - [Pneumonectomy and resection-anastomosis in malignant tumors of the lung. Problems of surgical radicalism and postoperative physiopathology]. PMID- 3050607 TI - [Leiomyosarcoma of the duodenum. Description of a clinical case]. PMID- 3050608 TI - [Malherbe's calcifying epithelioma. Apropos of 6 cases found in childhood]. PMID- 3050609 TI - [For a differentiated treatment of colorectal villous tumors]. PMID- 3050610 TI - [Pharmacological treatment of obesity. Drugs with predominantly anorectic action]. PMID- 3050611 TI - [Pirenzepine in the prevention of gastric intolerance to drugs used in pulmonary pathology]. PMID- 3050612 TI - [Effects of Saccharomyces cerevisiae in patients with intestinal dysbacteriosis]. PMID- 3050613 TI - [Pathology of duodenogastric reflux in patients with acute cranio-cerebral injuries. I. Various aspects of pathogenetic correlates and clinico-diagnostic problems]. PMID- 3050614 TI - [Pneumocystosis]. AB - On the basis of personal experience, the microbiological, epidemiological, clinical and therapeutic features of Pneumocystis carinii pneumonia are analysed. PMID- 3050615 TI - [Lymphadenopathies and diseases with manifestations of autoimmunity]. AB - The diseases responsible for lymphadenopathies with autoimmune features were examined. Such features play a major role in a vast number of clinical conditions whose aetiology is for the most part unknown but which present several clinical, histological and laboratory aspects in common. The most significant of these conditions are serum sickness and similar pictures induced by drugs, iodised contrast media and hymenoptera venom, angioimmunoblastic lymphadenopathies, giant multicentric lymph node hyperplasia, diffuse connectivitis, Wegener's granulomatosis lymphoid granulomatosis, necrotic lymphadenitis without granulocyte infiltration, mucocutaneous lymph node syndrome, angiofollicular hyperplasia with eosinophilia and histiocytosis of the sinuses. In some of these conditions, the lymphadenopathy is a constant and characteristic feature of the disease; in others it is common, in others rare and at time purely local. The autoimmune changes encountered in these conditions may, at times, be responsible for the morbidity, as in serum sickness. In others they merely constitute major or minor symptoms. In some they are no more than marginal aspects. The onset is usually acute with widespread symptoms and a clinical control featuring manifestations of hypersensitivity and immune deficiency. The most common and characteristic laboratory findings are polyclonal hypergammaglobulinaemia, circulating immune complexes, cryoglobulinemia and hypocomplementaemia and finally medullary plasmocytosis. Lymph node biopsy also tends to reveal a standard picture characterised by polymorphic infiltration of immunologically normal cells and the proliferation of newly formed small calibre blood vessels, mostly venules: In other words an aspecific reactive picture. For this reason diagnosis will sometimes be provided by blood tests, sometimes by repeated biopsy at a later date, sometimes by the evolution of the clinical picture. One other feature of these conditions and common, incidentally, to almost all diseases involving immunological alterations, is that they may be complicated by the appearance of lymphomas. Among the diseases quoted particular attention was paid to angioimmunoblastic lymphadenopathy and Castleman's disease given their greater frequency and the importance of their aetiopathogenic and clinical aspects. PMID- 3050616 TI - [Comparative study of defibrotide and calcium heparin in the prevention of postoperative deep venous thrombosis. A randomized multicenter study]. AB - The efficacy and tolerability of defibrotide (800 mg/i.v.) and calcium heparin (15,000 UI/s.c.) in the prophylaxis of post-surgical deep venous thrombosis (DVT) and pulmonary embolism (PE) were compared in a multicentre trial involving 60 Italian surgical institutions (general surgery, obstetrics and gynecology, urology). Total enrollment was 2,250 patients (defibrotide: 1.194; calcium heparin 1.056). According to the protocol, the clinical suspicion of DVT and/or PE led to in-depth diagnostic evaluations (DVT: Doppler ultrasound velocimetry; PE: chest X-rays; ECG, pulmonary scintigraphic scanning). The incidence of post surgical DVT was similar in the two groups (defibrotide: 8 patients; calcium heparin: 10 patients). A trend towards a lower incidence of DVT in the defibrotide group no PE; calcium heparin: 4 cases (chi 2 = 4.530, p less than 0.05). The local and systemic tolerability of both treatment was excellent. This trial, carried out in routine surgical practice, establishes the profibrinolytic approach to DVT prophylaxis as a sound and effective alternative to the traditional interference with the coagulation cascade. PMID- 3050617 TI - [Clinical pharmacology of non-steroidal anti-inflammatory drugs. A new benzothiazine derivative: cinnoxicam]. PMID- 3050618 TI - [New therapeutic prospects in gynecologic inflammation: nimesulide]. PMID- 3050619 TI - [Pascal's law and pelvic torque in the physiological mechanism of childbirth]. PMID- 3050622 TI - Diploma programs in nursing accredited by the NLN 1988-89. PMID- 3050620 TI - [Experience with three types of IUD (Gravidard, ML Cu 250, Nova T) in 950 women in a 24-month follow up study. Evaluation of side effects, complications and histomorphological study of endometrial changes induced]. PMID- 3050624 TI - Biological activities of chlorophyll derivatives. PMID- 3050621 TI - Enkephalin analogues depress synaptic potentials in rat dentate granule cells recorded intracellularly in vitro. AB - Enkephalin analogues were superfused onto hippocampal slices during intracellular recording of dentate granule cells. The enkephalins elicited either weak depolarizations, weak hyperpolarizations, or no effect on membrane potential, in about equal numbers of cells tested. Similarly, input resistance either decreased, increased or did not change, and was not well correlated with the potential changes. However, at all concentrations tested (2-10 microM) the enkephalins reduced evoked depolarizing synaptic potentials by up to 70%. We speculate that a major function of endogenous enkephalin-containing fibers projecting to the dentate is to dampen afferent synaptic transmission. PMID- 3050623 TI - Steroid danger in Kearns-Sayre syndrome (KSS). PMID- 3050625 TI - A feasible solution. PMID- 3050626 TI - Evaluating benign breast disease. AB - Benign breast diseases are present in some form in nearly all women. A basic understanding of breast anatomy and physiology, underlying pathology and methods available for evaluating breast problems provides the ability to assess the client's breast complaint. Four common breast problems--fibrocystic disease, fibroadenoma, mammary duct ectasia, and intraductal papilloma--are described along with guidelines for assessment and referral or follow-up. Current breast imaging options are discussed in terms of their usefulness and limitations. The value of consumer education regarding benign breast problems cannot be overstated. The National Cancer Institute offers free consumer information explaining benign breast diseases and evaluation techniques. PMID- 3050627 TI - Physical conditioning in the aging adult. AB - The benefits of exercise are well established but the impact of exercise on the older adult is not as clearly defined. Increasing numbers of people in the United States are over 65 years old. Thus health care providers will find themselves often involved with the older adult and will need an understanding of exercise in this age group. This article reviews the known benefits of exercise for the older adult, exercise-related age changes, and how to prescribe exercise for those 65 years and over. Selected case reports are used to provide examples of how specified guidelines can be incorporated into practice. PMID- 3050629 TI - Biological basis of the behavior of sick animals. AB - The most commonly recognized behavioral patterns of animals and people at the onset of febrile infectious diseases are lethargy, depression, anorexia, and reduction in grooming. Findings from recent lines of research are reviewed to formulate the perspective that the behavior of sick animals and people is not a maladaptive response or the effect of debilitation, but rather an organized, evolved behavioral strategy to facilitate the role of fever in combating viral and bacterial infections. The sick individual is viewed as being at a life or death juncture and its behavior is an all-out effort to overcome the disease. PMID- 3050630 TI - S-adenosylmethionine in the treatment of depression. AB - The antidepressant property of S-adenosylmethionine (SAMe) has been supported by several uncontrolled and controlled studies. Compared to standard antidepressant agents, SAMe has fewer side-effects and shorter lag period. Future studies to delineate SAMe-responsive depression are warranted. PMID- 3050628 TI - Behavioral performance effects of antihypertensive drugs: human and animal studies. AB - Antihypertensive drug treatments have been reported in clinical investigations to produce adverse effects to a degree that causes hypertensive patients to discontinue medication. Many of the debilitating effects reported by patients appear to be of central nervous system origin, such as sedation, fatigue, memory loss and sensorimotor disturbances. Human and animal laboratory studies in the past two decades have been characterizing the psychotropic effects of antihypertensive medications with use of a wide range of behavioral techniques. Antihypertensive drug classes covered in this review are beta-adrenergic blocking agents, alpha-adrenergic agonists, diuretics, angiotensin-converting enzyme inhibitors and calcium channel blockers. While findings in animal studies show generally greater behavioral impairments after administration of alpha-adrenergic agonists in comparison with other drug classes, the few laboratory studies conducted with hypertensive subjects present a confusing picture. A need for further laboratory research with hypertensive subjects and, study of antihypertensive drug combinations is discussed. PMID- 3050631 TI - Testicular hormones reduce individual differences in the aggressive behavior of male mice: a theory of hormone action. AB - A chronology of theoretical development in studies of the role of testicular hormones in murine aggression is presented. Evidence which brings into question the generality of current theory is reviewed, and the implications of this evidence for the direction of future research are discussed. A new method by which to characterize individual differences in aggressive behavior is described, and recent data which provide the basis for the development of a new theory are presented. It is theorized that testicular hormones reduce individual differences in the aggressive behavior of male mice, and that this behavioral "homogenization" is mediated by a testicular-hormone regulated "discrimination mechanism," possibly localized within the mouse olfactory system. The generality of this theory and the implications for other hormone/behavior systems are also discussed. PMID- 3050633 TI - Alcohol dependence and its medical consequences. PMID- 3050632 TI - Renal transplantation experience. A historical review and analysis. PMID- 3050634 TI - Temporary insanity and premenstrual syndrome: medical testimony in an 1865 murder trial. PMID- 3050635 TI - Recovery of beta-cell function postpartum in a patient with insulin-dependent diabetes mellitus. PMID- 3050636 TI - Asthma in New Zealand: some answers, more questions. PMID- 3050638 TI - Jeremy Bentham's head. PMID- 3050637 TI - A strategy for the control of malignant melanoma in New Zealand. PMID- 3050640 TI - Cadaver kidney retrieval for transplantation in Canterbury and Westland: 1972 1987. AB - In Canterbury and Westland, 106 cadaver kidney donors have provided 186 kidneys, which have been transplanted or submitted for transplantation. The overall rate of 16 donor/million population/year, since 1973, is high by international standards [1,2]. Male donors (68%) had a median age of 20 years, and females a median age of 24 years. Motor vehicle accidents were responsible for 58% of deaths, followed by subarachnoid haemorrhage (15%). Cadaver nephrectomy was performed outside normal working hours in 58% of cases. Overall, 10% of kidneys removed were discarded, but none have been discarded since September 1980. Anatomical anomalies were common (42%). PMID- 3050641 TI - Dr. JP FitzGerald: pioneer colonial surgeon, 1840-1854. PMID- 3050642 TI - Outpatient drugs: Medicare's new benefit. PMID- 3050643 TI - Bridging the gap: theory to practice--Part II, Research applications. PMID- 3050644 TI - Composites: yesterday, today and...?...tomorrow. PMID- 3050639 TI - Acute streptococcal necrotising fasciitis. AB - Two cases of acute streptococcal necrotising fasciitis are reported. Both patients were taking nonsteroidal antiinflammatory drugs when they developed this infection. Urgent surgical debridement was undertaken and resulted in a successful outcome in both patients. The clinical and histopathological features of this condition are reviewed. PMID- 3050649 TI - Measurement of absolute left ventricular volume by radionuclide angiography: a technical review. AB - Absolute left ventricular volumes have important clinical implications in the evaluation of cardiac performance. Several invasive and noninvasive techniques have been reported, none of which can be considered ideal for this purpose. Contrast angiography, echocardiography and radionuclide ventriculography are open to criticism. Different radioisotopic approaches are described with emphasis on the importance of accurate separation of left ventricular activity, the selection of background activity, and the correction for photon attenuation by body tissues. Improper use of statistics and validation techniques have obscured the value of these techniques. In the absence of a 'gold standard' there should be a 'radioisotopic' left ventricular volume with established independent characteristics, repeatability and reproducibility by which new approaches can be judged. PMID- 3050647 TI - Glass ionomer cements: "current status and applications". PMID- 3050646 TI - The etched porcelain veneer technique. PMID- 3050648 TI - Current composite resin techniques. PMID- 3050645 TI - Adhesion of bonded restorations after chemo-mechanical treatment of dentin. PMID- 3050650 TI - Background in 99Tcm DTPA renography evaluated by the impact of its components on individual kidney glomerular filtration rate. AB - Following injection for renography, 99Tcm-labelled diethylenetriamine-pentacetic acid (DTPA) rapidly enters the extravascular space. Background therefore comprises two components, a falling intravascular signal and an extravascular signal which initially rises. We estimated the relative magnitudes of these two components in terms of their impact on the calculation of differential renal function and individual kidney glomerular filtration rate (IKGFR) from the second phase of the 99Tcm-labelled DTPA renogram in 56 paediatric kidneys. We expressed each of the two background signals as a GFR equivalent. The GFR equivalent of the intravascular signal recorded from a peri-renal background region of interest (ROI), scaled by a factor equal to the ratio of the pixel numbers in the renal and background ROIs, was -39 (S.D. 14) ml min-1. The GFR equivalent of the extravascular signal was smaller than this and opposite to it at 23 (S.D. 10) ml min-1, giving a median ratio for the two equivalents of -1.68. Because of the opposing effects of the two background components on the second phase of the renogram, techniques recently described for the quantification of IKGFR from the renogram, and which eliminate the intravascular component, offer no theoretical advantage over a method of analysis which uses 'direct' subtraction of the total background signal. In practice, however, these new techniques are superior in their handling of 'noisy' data, consistently giving a lower coefficient of variation in their estimation of IKGFR. PMID- 3050651 TI - Peripheral placental separation: a review. AB - Marginal placental bleeding is a distinct entity, marked (usually) by minor degrees of vaginal bleeding and significantly associated with premature labor and premature rupture of the membranes. Pathologically it is characterized by an adherent marginal placental hematoma, which shows varying degrees of film deposition and leukocytic infiltration depending on the age of the clot. Marginal placental bleeding may occur only once or be repeated many times. It usually is not fatal to mother or fetus per se. The chief clinical significance of the marginal placental bleed is its tendency to be confused with placenta previa. The perinatal mortality associated with perpheral placental separation is largely that of prematurity. PMID- 3050652 TI - Delivery following prior cesarean section: an obstetrician's dilemma? PMID- 3050653 TI - The effects of human insulin on antibody formation in pregnant diabetics and their newborns. AB - We studied the immunogenicity of human insulin in 11 diabetic mothers and their newborns. Serum antibody formation was assayed by two different methods. Upon switching four patients from beef/pork insulin to human insulin, we found that elevated baseline antibody levels in three women decreased, in two to undetectable levels at term. The fourth patient had undetectable antibody levels at baseline and borderline levels at term. Only one of their four newborns had antibodies. Upon initiation of insulin treatment in another five diabetics without detectable antibodies at baseline, only two developed antibodies, and only one of their newborns developed antibodies. Two other patients, initially not on insulin, had baseline elevations of antibody that decreased with administration of human insulin; both of their newborns had antibodies. Overt diabetes evolved subsequently in both mothers after pregnancy. We conclude the following: 1) Upon transfer from beef/pork insulin to human insulin, mothers and their newborns show a decrease in insulin antibodies; 2) new patients initiated on insulin develop low levels of antibodies, if any, and their newborns also have low levels of antibodies if any; and 3) the decreased or absent immunogenicity of human insulin supports its use in pregnant diabetics. PMID- 3050654 TI - Clindamycin versus metronidazole in the treatment of bacterial vaginosis. AB - One hundred forty-three women with complaints of vaginitis were assigned to receive either 500 mg of metronidazole twice daily for 7 days or clindamycin 300 mg twice daily for 7 days. There was no significant difference in the failure rate between patients treated with clindamycin (6.1%) and those treated with metronidazole (4%). Adverse reactions were infrequent and mild in both treatment groups. Three patients who received clindamycin developed non-bloody diarrhea, which was mild and did not necessitate discontinuing therapy. We conclude that clindamycin may be a safe and effective alternative to metronidazole for treating women with bacterial vaginosis. PMID- 3050655 TI - Direct fluorescent antibody testing for endocervical Chlamydia trachomatis: factors affecting accuracy. AB - Endocervical swabbings obtained after two previous cleansing swabs from 202 women with indications for testing for genital chlamydial infection were evaluated for the presence of Chlamydia trachomatis by culture and two direct fluorescent monoclonal antibody tests. In comparison with culture, the two direct fluorescent antibody tests showed sensitivities of 37.5 and 56.5% and specificities of 97.0 and 99.4% when read by experienced microbiology technologists recently trained in chlamydia direct fluorescent antibody interpretation and blinded to culture results. Overall sensitivities of 69.6 and 78.3% for the direct fluorescent antibody tests were obtained by an expert interpreter during discrepancy analysis. When only direct fluorescent antibody test specimens from the first swab after endocervical cleansing were considered, recently trained interpreters obtained sensitivities of 53.8 and 69.2%, and both direct fluorescent antibody tests were 100% sensitive for the expert interpreter. These data emphasize the critical importance of observer expertise and swab order to the accuracy of chlamydia direct fluorescent antibody tests. Previous studies of these tests are examined to determine how these factors and others may have influenced the outcome. PMID- 3050657 TI - The menopausal syndrome. AB - The average life span of women living in the United States is approximately 80 years of age, with one-third of those years spent after menopause. Of the approximately 35-40 million postmenopausal American women, only 10% are currently using estrogen replacement therapy. Atrophy of the genitourinary system may be prevented or reversed with estrogen therapy. In addition, 75% of all postmenopausal women have vasomotor symptoms severe enough to lead them to seek medical consultation. For 65%, these symptoms persist for at least 1 year, and for 20%, the hot flushes are present for more than 5 years. Estrogen replacement therapy is the treatment of choice for the management of vasomotor symptoms, with or without additional behavior symptoms, and should be considered safe for most postmenopausal women. It protects the bones and, most important, the cardiovascular system. PMID- 3050658 TI - Determining "state-of-the-art" requires historical perspective. PMID- 3050659 TI - History and development of hysteroscopy. AB - The hysteroscope has evolved over the last two centuries. The first scientist to conduct light into the human body was Bozzini in 1805. The hysteroscope as we know it today is similar to the early cystoscope described and presented in 1877 by Nitze. The instruments and technique of endoscopy underwent advances by people such as Heineberg (1914), Rubin (1925), Mickulicz-Radecki (1927), and Norment (1949). Vulmiere applied cold light fiberoptics, and Marleschki (1965) introduced the early form of the contact hysteroscope. Edstrom and Fernstrom introduced the use of a solution of a high-molecular-weight dextran from beet sugar as a distending medium, and Hamou (1980) designed a 4-mm microcolpohysteroscope for histologic diagnosis in the reproductive tract. PMID- 3050660 TI - Future growth and development of hysteroscopy. AB - In less than two decades, hysteroscopy has evolved into a practical technique for the evaluation of the uterine cavity, with well-established indications such as evaluation of abnormal uterine bleeding and abnormal hysterograms as well as the treatment of intrauterine adhesions or the septate uterus, the removal of misplaced or embedded intrauterine devices, and the removal of submucous leiomyomas. As the use of hysteroscopy increases and more practitioners utilize it, new instrumentation and techniques are beginning to evolve, simplifying and facilitating not only a diagnostic examination, but also the more difficult and complex surgical interventions. Flexible endoscopes are being tested for possible use in the uterine cavity, and operative hysteroscopes with practical inflow and outflow accessory channels have been introduced. The accessory operative instrumentation has also been expanded and refined, and portable units for office examinations have been developed. The trend in the development of instrumentation is toward simplicity, effectiveness, and refinement in lenses with wider fields of view and accessory channels without compromising the total outer diameter of the endoscopes. As clinical applications expand as a result of increased use and proficiency in hysteroscopy, new applications undoubtedly will follow, such as closer study of the endometrium with and without additional magnification. This latter may permit a better understanding of the receptivity of the endometrium to the embryo, which may help in predicting successful nidations. Portable laser units with the capability to use the CO2 fiber undoubtedly will increase laser use through hysteroscopy, and in the near future, photodynamic therapy may become the best approach to the selective treatment of intrauterine lesions. The uterotubal junction will remain an attractive area for endoscopists to approach the fallopian tubes transcervically and eventually to accomplish tubal closure safely and effectively with the possibility of future reversibility. The hysteroscope will play an important role in new reproductive technologies, particularly those related to the gamete intrafallopian transfer via the uterine side and perhaps also for direct embryo transfers under visual control. The future of hysteroscopy thus is promising. The present diagnostic and therapeutic applications will not only become a standard of treatment but will expand as gynecologists will gain proficiency and confidence in this endoscopic method. PMID- 3050656 TI - The effects of postmenopausal estrogen therapy on the incidence of arteriosclerotic vascular disease. AB - Recent studies show that arteriosclerotic cardiovascular disease is associated with high low-density lipoprotein and low high-density lipoprotein cholesterol concentrations and, further, that reversal of these abnormalities can prevent cardiovascular disease. As postmenopausal estrogen therapy favorably changes low density lipoprotein and high-density lipoprotein cholesterol concentrations, it is hypothesized that reductions in cardiovascular disease will be observed in postmenopausal women so treated. The majority of at least 23 studies support this view. PMID- 3050664 TI - Hysteroscopic sterilization. AB - Hysteroscopic methods of tubal occlusion have not satisfied the criteria of an ideal contraceptive, although this technology still appears to be evolving. The aforementioned studies report the success and complication rates of very experienced hysteroscopists. A good potential exists for these techniques, but their widespread use will not come before greater experience and familiarity with the basics of hysteroscopy. PMID- 3050663 TI - Indications for hysteroscopy. AB - A description of current indications for hysteroscopy is presented, including a review of the literature. Emphasis is placed upon indications that are felt to be useful in the diagnosis and treatment of common gynecologic problems, including abnormal uterine bleeding, infertility, recurrent pregnancy wastage, and removal of intrauterine foreign bodies. PMID- 3050661 TI - Instrumentation in hysteroscopy. AB - The use of diagnostic and operative hysteroscopy has evolved rapidly over the past decade to become an integral part of gynecologic care. The hysteroscopies and ancillary equipment are readily available commercially, and the techniques can be mastered, as is true of any surgical technique, with an appropriate amount of supervised practice. With a good basic understanding of the types of equipment available and their appropriate uses, the maximum utility of the hysteroscopic technique can be fully realized in an environment of safety. PMID- 3050665 TI - Hysteroscopy as a diagnostic aid. AB - Recent improvements in instrumentation and technique have rendered hysteroscopy safe, simple, and effective in detecting a wide variety of pathologic conditions of the uterus, and the accuracy of the diagnostic process can be greatly enhanced by its use. However, hysteroscopy should not, by any means, be considered the ultimate diagnostic test. Rather, it should be applied judiciously and the information obtained combined with that from other methods, increasing the diagnostic accuracy. PMID- 3050666 TI - Laser ablation of the endometrium. AB - In summary, the Nd:YAG laser can be used safely and successfully to help women with menorrhagia avoid a hysterectomy. This is important not only for those women who medically are poor candidates but also for those women who for personal reasons do not want to lose their uterus or whose lifestyle will not allow them the time for a hysterectomy. Two basic applications of the laser to the endometrium are the touch and nontouch techniques. Although the nontouch technique appears to carry less risk of fluid overload and postoperative bleeding, it apparently does not decrease menstrual flow as much. Nevertheless, all techniques will allow most patients to avoid hysterectomy. PMID- 3050667 TI - Hysteroscopy in the infertile woman. AB - In summary, hysteroscopy is a useful diagnostic method in the evaluation of the uterine cavity. Hysteroscopy can localize the uterine lesions and determine their extent with more accuracy than the hysterosalpingogram. However, the meaning of some small lesions and their relation to infertility remains to be determined. Hysterosalpingography should be the first step in the evaluation of the uterine cavity and the tuboperitoneal factor in an infertile woman. Hysteroscopy should be combined with laparoscopy when any lesion is suspected on the hysterogram or when no other cause of infertility has been found. The diagnosis of uterine lesions can be followed by surgical treatment under laparoscopic guidance. PMID- 3050662 TI - Office hysteroscopy. AB - Several forces are acting on hysteroscopic surgeons to promote the 100-year-old practice of office hysteroscopy. It remains the surgeon's responsibility to triage patients properly to the office or hospital, and it is hoped that the principles discussed herein are helpful in that thinking. Office hysteroscopy implies more adaptability in the surgeon than in the surgeon's instruments. The indications, contraindications, and technique of office hysteroscopy do not differ significantly from those of hysteroscopy in the hospital on an awake patient. With current economic forces, instrumentation, and surgeon interest and training, office hysteroscopy will probably grow in popularity over the next several years. PMID- 3050668 TI - [Polypeptide growth factors in animal embryogenesis]. AB - Data about polypeptide growth factors in animal cells during embryogenesis and about the sensitivity of these cells to regulatory effect of the factors have been systematically reviewed. The conclusion was drawn that they play an important role in molecular mechanisms controlling cell proliferation and differentiation at the early embryonic stages. Information has also been provided about transforming growth factors and about the products of some proto-oncogenes, which are detected in the embryonic cells in amounts comparable with those in the transformed cells. However, this does not lead to malignant growth. Moreover, microenvironment of an intact embryo exerts an antitransforming influence on tumor cells. Studies of the "normalizing" effect of the embryonic cells may help in developing new methods of suppression of the malignant growth. PMID- 3050669 TI - [The biological action spectrum of the secretory products of the human B-cell lymphoblastoid line RPMI-641Ot]. AB - Secretory products of the lymphoblastoid line of human B-cells RPMI-6410t have a wide spectrum of biological activity. These products exert a growth-stimulating effect on B-cells of the 6410t strain obtained from the peripheral blood of a patient with acute myeloblastic leukemia and cytotoxic and cytostatic effects, respectively, on B-cells of the Raji and P3HR-I.G5 lines obtained from patients with Burkitt's lymphoma. They also exert a cytotoxic effect on murine L-cells. Biological activity is relieved at 70 degrees and at low and high pH. PMID- 3050670 TI - [Precursor cells of newly formed connective tissue in inflammatory foci in various organs in mice]. AB - The origin of fibroblast-like cells of the capsule around a foreign body in the spleen, liver, peritoneal cavity, subcutaneous connective tissue of mice, the localization of cells-precursors, their proliferative potencies and the ability to migrate through blood were studied using 3H-thymidine autoradiography. Precursors of the inflammation focus cells (irrespective of localization) reproduce intensively outside the limits of intraorganic connective tissue, supposedly, in hemopoietic organs of the bone marrow type and migrate, through the blood channel, into tissues (inflammation foci), where they terminate their differentiation. PMID- 3050671 TI - Diagnosis and treatment of chronic postoperative bacterial endophthalmitis. AB - Chronic postoperative bacterial endophthalmitis has recently assumed a prominent role in differential diagnosis of inflammation following extracapsular cataract extraction with posterior chamber intraocular lens implantation. The optimal diagnostic and therapeutic approach to this entity has not yet been clearly defined. We present a case of chronic postoperative Propionibacterium acnes endophthalmitis in which the diagnosis was made by anterior chamber paracentesis, and topical, periocular, and systemic antibiotic therapy resolved the inflammation. Anterior chamber paracentesis for aerobic and anaerobic cultures may be an appropriate initial diagnostic step in suspected cases. While successful treatment may require surgical intervention in some cases, others may respond to antibiotic therapy alone. PMID- 3050673 TI - Posterior chamber lens implantation in the absence of posterior capsular support. PMID- 3050672 TI - A technical variation of posterior chamber IOL implantation. AB - In posterior chamber intraocular lens (IOL) implantation, one of the most delicate phases is placing the loops in the capsular bag or in the ciliary sulcus. We describe a technique to simplify the phase of IOL insertion by tying the superior loop with a nylon suture. No changes to the lens and no special instruments are required. PMID- 3050674 TI - Electrophysiology and metallosis: support for an oxidative (free radical) mechanism in the human eye. AB - Nine cases of metal intraocular foreign body are presented. This study investigates the use of electrodiagnostic techniques in the diagnosis of metallosis and as a prognostic indicator. The electro-oculogram (EOG) and the electoretinogram (ERG) indicate that there are two mechanisms of metallosis, one of which involves cytotoxic damage by siderosomes, and the other lipid peroxidation of photoreceptor outer segments and retinal pigment epithelium. The evidence suggests that the location of the foreign body in the eye determines whether either or both of these mechanisms occur. We propose that these tests can help the ophthalmologist decide when a foreign body has to be removed and give information as to the mechanism of the pathological process. PMID- 3050675 TI - Chronic vitritis with macrophagic inclusions. A sequela of treated endophthalmitis due to a coryneform bacterium. AB - A 75-year-old woman was treated successfully for endophthalmitis due to a coryneform bacterium contracted from a contaminated corneal graft. We were able to study the involved eye histologically when the patient died unexpectedly 5 1/2 weeks after treatment. The vitreous contained a moderate number of macrophages filled with PAS-positive particles. Ultrastructurally, the PAS-positive particles corresponded to degenerating bacterial cell walls. The striking resemblance of the macrophages in this case to macrophages in Whipple's disease is intriguing because Corynebacterium has been the most frequently implicated bacterial genus in the pathogenesis of Whipple's disease. PMID- 3050676 TI - Epithelial abnormalities and sterile ulceration of epikeratoplasty grafts. AB - Six epikeratoplasty (EKP) specimens were obtained 1 to 10 months after refractive keratoplasty for aphakia (2 cases), keratoconus (1 case), and myopia (3 cases). The EKP grafts were removed because of postoperative complications such as delayed reepithelialization, lenticule edema, stromal ulceration, graft opacification, and inability to improve vision in a clear graft. The extent of epithelial abnormalities was associated directly with the severity of structural changes in the underlying basal lamina, Bowman's layer, and stroma as evidenced by electron microscopic examination. Epithelial defects were associated with varying degrees of sterile stromal ulceration of the underlying lenticule. In one case, an extensive full-thickness ulceration of the EKP button occurred in the area of persistent epithelial defects and was associated with a major ulceration of the underlying recipient's corneal stroma. Removing the failing button halted the ulcerative process with prompt reepithelialization. In the absence of inflammation and infection, the persistent epithelial abnormalities and defects leading to proteolytic degradation of the EKP buttons constitute an important mechanism of graft failure with major visual consequences and possibly irreversible ocular damage. PMID- 3050678 TI - A clinical trial of timolol and epinephrine in the treatment of primary open angle glaucoma. AB - Forty-seven patients with chronic open-angle glaucoma who required therapy but who had never been treated previously were enrolled in a prospective randomized, long-term clinical trial comparing the effectiveness of 0.5% timolol maleate and 1% epinephrine. Patients were followed for an average of 33 months. There were 20 failures, 18 of which were because of inadequate intraocular pressure (IOP) control (required 20% reduction in outflow pressure). Twelve failures were in the epinephrine group and eight were in the timolol group. During the first year of therapy, 35% compared with 0% of patients failed in the epinephrine and timolol groups, respectively (P less than 0.01). During the later years of the study, the failure rates in the two groups were similar. The results suggest that timolol is superior to epinephrine during the first year of therapy, and likely as effective as epinephrine in the long-term treatment of glaucoma. PMID- 3050677 TI - Indications and treatment of keratoconus using epikeratophakia. AB - Nineteen patients with keratoconus underwent epikeratophakia by one of the authors (DSD) and were followed from 3 to 29 months. Patient selection criteria included contact lens failure, and minimal or no central corneal scarring. Uncorrected visual acuity improved by three or more Snellen lines in 13 of 19 patients (68%). Postoperatively, after more than 6 months follow-up, 81% of the patients had best-corrected visual acuity of 20/40 or better. A mean flattening of 4.73 diopters (D) occurred on keratometry readings. There was a mean decrease in refractive cylinder of 2.84 D. Spherical equivalent refraction showed a mean decrease in myopia of 4.64 D. Five patients had postoperative refractive cylinder greater than 4 D requiring relaxing incision(s). With some patients having been followed for more than 2 years, no recurrences of keratoconus have been noted. In properly selected patients, epikeratophakia can effectively be used to treat keratoconus and thus avoid potential intraocular surgical complications and immunogenic phenomena. PMID- 3050681 TI - Glaucoma update. PMID- 3050682 TI - Keratoconus. Evaluation of recent trends in the surgical and nonsurgical correction of keratoconus. AB - Recent focus on the optical correction for "contact lens failure" keratoconus patients has been in the use of epikeratoplasty as an alternative to penetrating keratoplasty. In a 2-year retrospective review of keratoconus patients, 33 were referred as "contact lens failures" and as such might be considered candidates for surgical intervention. Of these, 29 patients (average keratometry, 50.4; range, 43-73) were successfully refitted with contact lenses. All patients obtained wearing times of 12 hours and achieved a visual acuity of 20/40 or better, with 85% realizing 20/30 or better. The data suggest that with concerted effort most keratoconus patients may be successfully refitted despite initial contact lens failure. PMID- 3050680 TI - Evaluation of the t test as a method of detecting visual field changes. AB - Automated perimetry is used to monitor changes associated with many ocular conditions such as glaucoma, neuro-ophthalmologic disorders, and retinal diseases. Comparisons among sequential visual fields are crucial to the determination of progression or regression of ocular anomalies, and to evaluate the efficacy of various therapeutic regimens. The Delta program for the Octopus perimeter uses a t test to provide an objective comparison of visual fields. To evaluate the utility of the t test for detecting visual field changes, the authors used computer simulation to generate pairs of visual fields with known alterations, and made comparisons with the t test procedure used by the Delta program. An initial visual field based on normal data was produced and stored. A second visual field, based on the same normal data but with a scotoma of known size and depth, was also produced and stored. A t test (as used by the Delta program) was then used to compare the two visual fields for statistically significant differences. Our results indicate that the t test was very sensitive to small changes of the entire visual field, such as those produced by both pathologic and nonpathologic (day-to-day sensitivity variations, changes in pupil size, etc.) factors. However, the t test was unable to reliably detect small to moderate scotomata (less than or equal to 18 degrees in diameter), even when the sensitivity loss was greater than 35 dB. These findings suggest that the t test has limited clinical utility for objective detection of pathologic visual field loss, and that additional procedures may be necessary to provide reliable detection of visual field changes. PMID- 3050679 TI - Ocular hypotensive efficacy of 0.25% levobunolol instilled once daily. AB - The authors evaluated the efficacy of once-daily treatment with levobunolol in patients with primary open-angle glaucoma or ocular hypertension in an open labeled, two-phase titration clinical trial. All patients started the study using 0.25% levobunolol administered once daily for 3 months (phase I). If a patient's intraocular pressure (IOP) was not controlled with this concentration of levobunolol, the concentration was increased to 0.5% administered once daily for 3 months (phase II). During phase I, a significant reduction in IOP was observed in 21 of the 29 patients (72%), with an average IOP reduction of 24%. During phase II, in six patients whose IOP was reduced inadequately with 0.25% levobunolol, one had a significant reduction in IOP with 0.5% levobunolol. The authors concluded that levobunolol, instilled once daily at a concentration of 0.25%, was effective in significantly reducing IOP in the majority of the patients evaluated. PMID- 3050683 TI - Linear IgA disease. The ocular manifestations. AB - The ocular history and examination of a 54-year-old Filipino woman with linear IgA disease is described. Results of the eye examination were consistent with chronic cicatricial conjunctivitis, showing subconjunctival fibrosis and symblepharon formation. Direct immunofluorescence of the conjunctiva was positive for IgA and C3 in a linear pattern along the epithelial basement membrane. The ophthalmologic and dermatologic findings in linear IgA disease are compared with those of dermatitis herpetiformis, bullous pemphigoid, and cicatricial pemphigoid. This is the first documented case report of the ocular manifestations of linear IgA disease in the American literature. PMID- 3050686 TI - ALO 2145 reduces the intraocular pressure elevation after anterior segment laser surgery. AB - The effect of an alpha adrenergic agonist, ALO 2145 (para-amino-clonidine [PAC]), was examined in a double-masked, multicenter study on the postoperative intraocular pressure (IOP) elevation after laser surgery in 165 patients (83 trabeculoplasty, 36 iridotomy, and 46 capsulotomy). One drop of 1.0% ALO 2145 or vehicle was instilled 1 hour before and immediately after laser surgery. The mean IOP increase in the ALO 2145-treated group was lower (P less than 0.05) than in the placebo group at each of the first three postoperative hours. Overall, 18% of placebo-treated eyes experienced IOP increases greater than or equal to 10 mmHg as compared with 4% of ALO 2145-treated eyes (P less than 0.003). Ocular and systemic side effects were minimal and did not differ between the treatment groups. ALO 2145 safely and effectively reduced the incidence and magnitude of potentially harmful IOP elevations after anterior segment laser surgery. PMID- 3050685 TI - Choroidal melanoma with pigment dispersion in vitreous and melanomalytic glaucoma. AB - A 39-year-old black man underwent enucleation of the left eye because of poor vision, ocular pain, and intractable glaucoma secondary to a choroidal tumor. Two diagnostic vitrectomies, performed 11 and 7 months before enucleation, had failed to disclose the proper diagnosis. Histologic diagnosis was necrotic malignant melanoma of the choroid with melanocytoma cells, extensive pigment dispersion throughout the eye, and melanomalytic glaucoma. Possible mechanisms of tumor necrosis are reviewed. PMID- 3050684 TI - Molteno implant for control of glaucoma in eyes after penetrating keratoplasty. AB - Seventeen patients (17 eyes) underwent implantation of a single plate Molteno implant for medically uncontrollable intraocular pressures after penetrating keratoplasty. Most of the eyes had extensive peripheral anterior synechiae, and 16 of 17 (94%) were pseudophakic or aphakic following keratoplasty. Other glaucoma procedures had been performed previously on 13 eyes: argon laser trabeculoplasty (one eye), trabeculectomy (seven eyes), transpupillary argon laser cyclophotocoagulation (three eyes), and cyclocryotherapy (three eyes). Follow-up ranged from 5 to 28 months (mean, 13 months). Three eyes underwent repeat Molteno implantation when intraocular pressure (IOP) was not satisfactorily reduced after the first procedure. Considering one eye with chronic hypotony as a failure, 12 of 17 eyes (71%) had IOPs of less than 21 mmHg at the time of the three most recent postoperative examinations after a single Molteno implant. Repeat implants in three eyes increased the number of eyes with IOPs of less than 21 mmHg to 14 (82%). Corneal allograft rejection after Molteno implantation occurred in seven eyes; two of these were successfully reversed with corticosteroid therapy. Three of the five eyes with irreversible graft rejection were regrafted, and two of these grafts have remained clear. Including the regrafted eyes, 13 eyes had clear grafts and controlled IOPs at the most recent postoperative examination. The Molteno implant may prove useful in the management of medically uncontrollable glaucoma following penetrating keratoplasty; however, there appears to be a substantial risk of postoperative graft rejection. PMID- 3050687 TI - Hemangioblastoma of the optic nerve. Report of a case and review of literature. AB - The authors present a case of an optic nerve hemangioblastoma in a young woman with von Hippel-Lindau disease. The initial diagnosis was made by incisional biopsy. Tumor growth led to progressive proptosis and loss of light perception. Excision was carried out by lateral orbitotomy. Clinically and radiographically, the tumor resembled an optic nerve meningioma or glioma. Review of the other known cases offers no information as to the potential spread of this benign tumor from the intraorbital optic nerve to the optic canal. Optic nerve hemangioblastoma must be considered in the differential diagnosis of optic nerve tumors in patients with or without von Hippel-Lindau disease. PMID- 3050688 TI - Immunodiagnosis of adult chlamydial conjunctivitis. AB - This study presents data from a prospective comparison of four currently available diagnostic tests for Chlamydia trachomatis infection. Seventy-six patients clinically suspicious for chlamydial conjunctivitis were all tested with Giemsa stain cytology, direct monoclonal fluorescent antibody (DFA) microscopy, enzyme immunosorbent assay (EIA) for chlamydial antigens, and standard McCoy cell culture. When compared with primary cell culture, diagnostic Giemsa inclusions had a sensitivity and specificity of 43 and 100%, respectively, supportive Giemsa cytology 71 and 67%, the enzyme immunoassay 71 and 97%, and the monoclonal fluorescent antibody 57 and 81%. Each nonculture method has distinct advantages in terms of cost, technical difficulty, speed, and accuracy, which dictate selection of the most appropriate test for office or laboratory diagnosis of chlamydial conjunctivitis. PMID- 3050689 TI - Episodic conjunctival inflammation after Stevens-Johnson syndrome. AB - The authors studied the histopathologic, ultrastructural, and immunopathologic characteristics of conjunctiva from patients with Stevens-Johnson syndrome (SJS). A small subset of SJS patients with recurrent conjunctival inflammation unassociated with external factors such as lid margin keratinization, sicca syndrome, trichiasis, or entropion was identified. The ultrastructural and immunopathologic characteristics of the conjunctiva from these patients were distinctly different from those of the conjunctiva from SJS patients without recurrent conjunctivitis, and suggested an active, immunologically mediated inflammation. Vasculitis or perivasculitis, immunoreactant deposition in vessel walls, vascular basement membrane disruption, thickening, and reduplication, and a preponderance of helper T-lymphocytes, macrophages, and Langerhans' cells were the notable distinguishing features in those patients with recurrent conjunctival inflammation. This rare clinical syndrome may represent the ocular counterpart to recurrent dermal or oral mucosal erythema multiforme. PMID- 3050690 TI - Inflammatory diseases of the peripheral cornea. AB - Immunologic differences exist between the peripheral and central cornea. The peripheral cornea is closer to the conjunctiva, which has all of the immunologic machinery necessary to generate an immune response. The peripheral cornea has more Langerhans' cells and IgM than the central cornea. The peripheral cornea also has more C1, the recognition unit of the classic pathway of complement, than the central cornea so that antigen-antibody complexes, whether formed in the cornea itself or whether derived from the tears, aqueous humor, or limbal vessels, may activate complement more effectively in the peripheral than central cornea. Autoimmune diseases that involve the peripheral cornea include Mooren's ulcer and collagen vascular diseases. The humoral- and cell-mediated autoimmune phenomena that are associated with Mooren's ulcer and its response to immunosuppressive therapy suggest that it is an autoimmune disease directed against the cornea itself. Collagen vascular diseases may be associated with peripheral corneal ulcers with or without scleritis. In these diseases, circulating immune complexes may lodge in the limbal vasculature causing an immune vasculitis or deposit in the peripheral cornea setting off the complement cascade. Peripheral corneal diseases that probably represent a hypersensitivity reaction to exogenous antigens include catarrhal infiltrates and ulcers and phlyctenules. In the United States today, these corneal lesions are generally associated with staphylococcal blepharitis. Experimental models suggest that hypersensitivity to Staphylococcus aureus cell wall antigens may be important to their immunopathogenesis. PMID- 3050691 TI - Botulinum toxin A-induced protective ptosis in corneal disease. AB - Botulinum toxin A produces a temporary, flaccid ptosis when injected into the levator palpebrae superioris muscle. The resulting protective ptosis was used to aid healing in 21 cases of indolent ulceration, and, prophylactically, in 4 cases of neuroparalytic keratitis. Of the indolent ulcers, 90% healed completely. In all but one case, the cornea was covered completely by the lid and complete ptosis was produced in 75% of cases in an average of 3.6 days, lasting for 16 days on average before recovery began. Recovery of levator function was complete in 8.5 weeks on average. Superior rectus underaction was seen in 68% of cases but this recovered completely in all cases in an average of 6 weeks. Impression cytology showed a trend toward normal conjunctival morphology as healing progressed. PMID- 3050692 TI - Cataract surgery in ocular cicatricial pemphigoid. AB - The authors report the results of their experience with cataract surgery in 20 patients (26 eyes) with biopsy-proven cicatricial pemphigoid. All patients were on systemic immunosuppression at the time of surgery (dapsone, azathioprine, cyclophosphamide, or combinations) and were treated with perioperative oral corticosteroids. Patients were evaluated pre- and postoperatively for conjunctival inflammation, conjunctival cicatrization, degree of keratopathy, and disease stage. No patient progressed in disease stage. Vision improved an average of 3.5 Snellen lines (-3 to +8). Worse outcome was associated with chemotherapy intolerance or the presence of any preoperative conjunctival inflammation. Thirteen patients remained on immunosuppressives for the entire study. Corneal ulcers developed postoperatively in three patients in whom continued immunosuppression was not tolerated. Possible mechanisms for inflammatory exacerbation after surgery are discussed. Results indicate that after successful abolition of all conjunctival inflammation through chemotherapy, cataract surgery may be safely performed in patients with cicatricial pemphigoid. PMID- 3050693 TI - Keratoconus. Contact lens or keratoplasty? AB - The success rate of contact lens fitting and rate of progression to keratoplasty were evaluated for 115 consecutive patients with keratoconus. Of 190 nonoperated eyes that needed to be fit, 25 (13%) could not be fit, whereas 165 eyes (87%) could be fit. Most of these eyes had been referred for keratoplasty after previous contact lens fittings had no longer been successful. Of the 165 eyes that could be fit, 51 (31%) ultimately needed keratoplasty after an average of 38.4 months of lens wear, and 114 eyes (69%) did not require keratoplasty over an average follow-up interval of 63 months of wearing contact lenses. The average initial keratoplasty reading in these two groups was 56.8 and 51.8 diopters (D), respectively. Special design, bispheric lenses were required in 125 of these 165 eyes (76%) and frequent lens changes were necessary. Of 88 postoperative eyes, 53 (60%) wore contact lenses for best vision. Keratoplasty can be delayed or avoided in many keratoconus patients by using contact lenses, especially special design, bispheric lenses. Also, keratoconus eyes often need contact lenses after keratoplasty. PMID- 3050694 TI - Excimer laser trephination in penetrating keratoplasty. Morphologic features and wound healing. AB - The imprecision of trephination of donor and recipient corneas is a major factor in post-keratoplasty astigmatism. In order to improve the quality of trephination, the authors developed a rotating slit delivery system for noncontact penetrating keratoplasty trephination using the excimer laser at 193 nm. Scanning electron microscopy (SEM), transmission electron microscopy (TEM), and light microscopy (LM) demonstrated the superior quality of excimer-cut buttons and recipient beds as compared with those obtained by free hand and suction trephines in human cadaver and rabbit eyes. The laser trephined more regularly and precisely without distortion of corneal topography and with less damage to adjacent corneal tissue. The authors morphologically examined wound healing at 6 hours, 12 hours, 3 days, 5 days, 2 weeks, 2 months, and 3 months after penetrating keratoplasty with laser and mechanical trephination in an animal autograft model. The laser did not cause any adverse alteration of wound healing processes including cellular migration, proliferation, and production of new tissue. PMID- 3050695 TI - The nationwide study of epikeratophakia for aphakia in older children. AB - A nationwide study of epikeratophakia for aphakia in older children was conducted from March 1984 to March 1986. Sixty-three patients, 8 to 18 years of age, underwent this procedure in 65 eyes. Twenty-eight patients had congenital cataracts and 35 had traumatic cataracts. Fifty-one of the 65 eyes were aphakic at the time of surgery (secondary procedures). All surgeries were successful; no tissue lenses were lost or removed. Postoperatively, 73% of the patients were within 3 diopters (D) of emmetropia. The patients with congenital cataracts gained an average of one Snellen line of best-corrected visual acuity; patients with traumatic cataracts lost an average of one Snellen line of best-corrected visual acuity. In older pediatric patients, epikeratophakia appears to be a safe and effective procedure for the correction of aphakia. PMID- 3050696 TI - Edward Jackson, MD--a historical perspective of his contributions to refraction and to ophthalmology. AB - Edward Jackson died October 29, 1942, at 86 years of age. He served as president of the major national ophthalmologic organizations and was professor and chairman of the Department of Ophthalmology at the University of Colorado. He established the first graduate course for ophthalmologists, suggested the formation of the instruction courses of the American Academy of Ophthalmology, and was the principal founder of the American Board of Ophthalmology. He founded and edited the Yearbook of Ophthalmology and Ophthalmic Literature. In 1918, he became editor of the third series of the American Journal of Ophthalmology, which consolidated five ophthalmic periodicals. In 1885, he popularized retinoscopy in the United States and was mainly responsible for making it a practical refraction tool. Two years later, he described the cross cylinder to determine the presence or absence of astigmatism. In 1907, he described the use of the cross cylinder to refine the axis of a correcting cylinder in astigmatism. He lectured and wrote widely and published over 700 scientific articles, book chapters, and books. PMID- 3050697 TI - Relationship between sympathetic ophthalmia, phacoanaphylatic endophthalmitis, and Vogt-Koyanagi-Harada disease. AB - The more than coincidental occurrence of phacoanaphylatic endophthalmitis (PE) in sympathetic ophthalmia, and the similarity of the dissimilarity between Vogt Koyanagi-Harada (VKH) disease and sympathetic ophthalmia have been well described both clinically and histopathologically. The etiology and pathogenesis of these three diseases are still not fully understood. Identifying and distinctive characteristics among them include the history of ocular trauma in sympathetic ophthalmia, rupture of the lens capsule in phacoanaphylatic endophthalmitis, and involvement of the skin, ear, and central nervous system in VKH disease. A T-cell mediated immune reaction to ocular antigens seems to play a major role in sympathetic ophthalmia and VKH disease. A B-cell-related Arthus reaction to lens antigen seems to be the principle mechanism of PE. Thus, these three diseases may represent a spectrum of uveitis. At one end is the delayed-type hypersensitivity disease of sympathetic ophthalmia, whereas at the opposite end is the immune complex disease of PE, with VKH disease in the middle of this uveitic spectrum. PMID- 3050700 TI - Chorioretinitis in the contralateral eye of a patient with Acanthamoeba keratitis. AB - Chorioretinitis developed in the right eye of a patient with contact lens associated Acanthamoeba keratitis in the left eye during an acute exacerbation of the keratitis. This chorioretinitis may have resulted from hematogenous dissemination from his corneal infection. PMID- 3050699 TI - Spontaneous decalcification of a choroidal osteoma. AB - A 23-year-old woman presented with a clearly defined, pale orange choroidal tumor superior to the right optic disc in 1976. The patient was followed. After a choroidal osteoma was first reported in 1977, this diagnosis was confirmed in this patient using ultrasonography and orbital tomography. The lesion grew very slowly over the next 5 years. In 1981, the choroidal osteoma began to thin and decalcify. Subretinal neovascularization developed in 1982 and was treated with argon laser photocoagulation. In 1983, the tumor was thinner and less calcified. During the next 18 months, it became completely decalcified and essentially disappeared leaving only a bed of pigment epithelial and choriocapillaris atrophy. This was confirmed with fluorescein angiography and ultrasonography. PMID- 3050698 TI - Primary intraocular lymphoma (ocular reticulum cell sarcoma) diagnosis and management. AB - The authors retrospectively reviewed the diagnosis and management of 20 intraocular lymphoma patients who initially presented with either ocular or central nervous system (CNS) disease. As the ophthalmic community has become more aware of this entity, the interval between symptoms and diagnosis has significantly shortened. Diagnosis can usually be made on cytopathologic examination of vitreous cells. However, in three cases more than one vitreous biopsy was necessary. Results of cytologic examination appeared to be more accurate than those of conventional lymphocyte surface marker studies in the diagnosis of an intraocular lymphoma. Long-term survival occurred in some patients treated with a combination of intrathecal chemotherapy and ocular/CNS irradiation. PMID- 3050702 TI - An evaluation of the photographs of William H. Crisp, MD. AB - The popularity of the Jackson cross cylinder is based on the assumption that its end points are "equal" to each other. Crisp's photographs, showing equal end points, popularized the Jackson cross cylinder technique. Photographs presented in this study show the anamorphic distortions of Snellen letters induced by the Jackson cross cylinder and suggest that Crisp's photographs were in error. His photographs do not represent the images seen by the patient when the Jackson cross cylinder refraction technique is used with Snellen letters. PMID- 3050704 TI - Temporary keratoprosthesis for combined penetrating keratoplasty, pars plana vitrectomy, and repair of retinal detachment. AB - The Landers-Foulks temporary keratoprosthesis was used to combine penetrating keratoplasty, pars plana vitrectomy, and scleral buckling in the management of 13 eyes with opaque cornea and posterior segment abnormalities. In seven cases, trauma precipitated the ocular disease. Complications of cataract surgery resulted in anterior and posterior segment pathology in six cases. The corneal graft was initially clear in all cases. However, corneal edema complicated phthisis bulbi in four cases and followed homograft reaction in two cases. Eight eyes with retinal detachment (RD) preoperatively were successfully reattached. In five eyes, the retina redetached as these eyes became phthisical. Visual function improved in six cases. In general, eyes with a history of trauma had a much poorer outcome than did eyes with anterior and posterior segment problems related to previous cataract surgery. PMID- 3050703 TI - The role of ultrasound in the management of retinopathy of prematurity. AB - The authors reviewed the ultrasonic appearance of 54 eyes (27 patients) with advanced retinopathy of prematurity (ROP) and compared the various retinal configurations with the findings at surgery. They found that ultrasound is a sensitive indicator of retinal configuration (type and degree of retinal detachment [RD]), as well as showing the existence of subretinal or choroidal hemorrhage often present in these severely affected eyes. Ultrasound is a useful modality to evaluate, document, and follow the progression of RD in ROP. PMID- 3050701 TI - Frozen section of the optic nerve in retinoblastoma surgery. AB - The usefulness of the frozen section examination to determine the extension of retinoblastoma into the optic nerve is discussed. Frozen sections performed at the time of surgery in seven retinoblastoma patients revealed the presence of tumor at the resection margin of the optic nerve in two cases. PMID- 3050705 TI - Histopathologic effects of ultrasonically induced hyperthermia in intraocular malignant melanoma. AB - Four cases of human intraocular malignant melanoma were treated with ultrasonically induced hyperthermia immediately before enucleation. Tumors were treated in two regimens: 30 minutes at 43 degrees to 45 degrees C and 5 minutes at greater than 50 degrees C. Temperatures were estimated from applied power levels, based on empirical data and mathematical models. Histopathologic changes observed in human tumors were compared with changes seen in malignant melanoma xenografts in athymic nude mice which were treated with ultrasonically induced hyperthermia for 30 minutes at 42 degrees to 46 degrees C. The effects of treatment were similar to changes seen in the animal model treated under analogous conditions: increased intercellular spacing, cytoplasmic vacuole formation, clumping of chromatin, breaks in cell membranes, and swelling and collapse of cells. Perivascular and peripheral zones sometimes showed decreased damage levels. The high temperature (greater than 50 degrees C) technique is presently being used as a means of "sterilizing" tumors before planned enucleation. The moderate temperature (43 degrees-45 degrees C) technique has been used in combination with radiotherapy to treat tumors when vision can be salvaged. PMID- 3050706 TI - Viruses and salivary gland disease: are there associations? AB - Viruses can cause sialadenitis and may be associated with other diseases of salivary glands, particularly immunologically mediated and neoplastic lesions. The evidence that such an association with Sjogren's syndrome is causal is reviewed here and shown to be fairly tenuous at present. PMID- 3050708 TI - Diabetic polyradiculopathy with trigeminal nerve involvement. A case report. AB - A 20-year-old white woman with controlled diabetes mellitus of 8 years' duration and a current diagnosis of diabetic polyradiculopathy had oral pain suggesting involvement of the mandibular branch of the trigeminal nerve and mimicking trigeminal neuralgia. Cranial nerve involvement in diabetic polyradiculopathy is rare, and trigeminal nerve involvement with pain has not been reported. PMID- 3050707 TI - Management of an oroantral fistula in a patient with Wilson's disease: case report and review of the literature. AB - After removal of an impacted maxillary third molar, an oroantral fistula developed in a patient with Wilson's disease. Management consisted of antibiotics, decongestants, irrigation, and surgical closure. Complications of treatment did not directly involve the disease but, rather, were related to the therapeutic agent penicillamine. Penicillamine causes interference between the cross links of tropocollagen molecules and cleaves newly formed molecules. Reduction in dosage is recommended when surgery is planned to increase collagen formation and, thus, healing. Such a measure was undertaken in this case. The patient healed uneventfully. A review of Wilson's disease and a case report are presented. PMID- 3050709 TI - Gingival manifestations of childhood cicatricial pemphigoid. AB - Cicatricial pemphigoid (CP) is found almost exclusively among middle-aged and elderly persons. This article describes a rare case of CP in a 14-year-old girl; it appeared on the mandibular anterior gingiva as desquamative gingivitis. Histologic examination of the lesions showed a subepithelial bulla. Immunofluorescence of gingival biopsy revealed immunoglobulin G, protein C3, and faint immunoglobulin A deposition along the basement membrane zone; results for normal skin were negative. Indirect immunofluorescence produced negative results. There are only six documented cases of childhood CP previously reported in the literature. The clinical and immunologic features of these cases are reviewed. PMID- 3050710 TI - "Otodental" dysplasia. AB - The case of a 3 11/12-year-old Chinese boy with the dental abnormalities of "otodental" dysplasia is reported. Hearing was normal. Dental anomalies consisted of delayed eruption of globe-shaped molars, bulbous deciduous canines, and double pulp chambers in the molars. Radiographs taken 4 years later showed taurodontic molars, supernumerary microdontic teeth, retarded formation of premolars, and probable aplasia of the mandibular second premolars. PMID- 3050711 TI - Chronic indolent orofacial herpes simplex virus infection in chronic leukemia: a report of three cases. AB - Orofacial mucocutaneous infections caused by herpes simplex virus (HSV) may exhibit a distinct chronic indolent pattern of behavior in some immunosuppressed patients as opposed to the more familiar aggressive patterns. Three patients with chronic leukemia who illustrate this chronic indolent pattern are presented. These cases should alert the clinician to the variable clinical appearance that HSV may adopt in the immunosuppressed patient. PMID- 3050712 TI - Evoked otoacoustic emissions: a fundamental and clinical survey. AB - Evoked otoacoustic emissions (EOAEs) hold some promise as a fast, objective and noninvasive procedure to study the cochlea at the outer hair cell level. This paper summarizes the fundamental aspects of EOAEs, in particular their relationship with the active biomechanical properties of the cochlea. The properties of EOAEs are analyzed in normally hearing ears, in ears with sensorineural hearing loss, especially in Meniere's disease, in acoustic neuromas and in central deafness; the properties of EOAEs were also evaluated as to their suitability for screening auditory dysfunction in infants. PMID- 3050714 TI - Mucosal immunology--with special reference to specific immune defence of the upper respiratory tract. AB - The mucosa of the upper respiratory tract is protected by a secretory immune system which is under complex immunoregulatory control. B cells with a potential for J-chain expression are initially stimulated in mucosa-associated lymphoid tissue (probably including the tonsils) and thereafter migrate through lymph and blood to glandular sites where they differentiate to immunoglobulin-producing immunocytes. Most locally produced immunoglobulin normally consists of dimeric IgA which is selectively transported through serous glandular cells by means of an epithelial receptor protein called the secretory component (SC). IgM is also subjected to SC-mediated transport. In patients with selective IgA deficiency, secretory IgA is lacking, but it may be satisfactorily replaced by protective secretory IgM. In other IgA-deficient patients, however, immunoregulatory compensation gives rise to a large number of IgD-producing cells in respiratory mucosae. IgD cannot act as a secretory antibody and these patients are prone to have recurrent infections. There are thus large individual variations in the secretory immune system, which in the future hopefully may be subjected to regulatory manipulation. PMID- 3050716 TI - Bilateral simultaneous Bell's palsy. Two cases following herpes simplex gingivostomatitis. AB - Bilateral simultaneous Bell's palsy (BSBP) is an infrequent clinical presentation. Two patients with BSBP following a recent herpes simplex gingivostomatitis are described. From a clinical point of view, these cases support the concept that Bell's palsy may be a partial manifestation of a generalized polyneuropathy, and most probably caused by a reactivated latent herpes simplex virus. PMID- 3050715 TI - Localization of IgE synthesis in immediate-type allergy of the upper respiratory tract. AB - The localization of allergen-specific IgE synthesis in allergic diseases of the upper respiratory tract is so far unknown. It has been suggested that the IgE production takes place in the nasal mucosa itself. The present immunohistochemical studies with anti-IgE and monoclonal markers for B lymphocytes, plasma cells, antigens of the major histocompatibility complex, T helper and T suppressor cells indicate that there are no IgE-producing plasma cells in the nasal mucosa of patients with seasonal allergic rhinitis. The IgE associated cells have been defined as two different types of mast cells. Furthermore we found IgE-associated lymphoid follicles in palatine as well as in nasopharyngeal tonsils and in cervical lymph nodes from allergic patients. IgE specific activated B lymphocytes and plasma cells were identified in direct contact with migratory mast cells in these lymphoid tissues. We therefore suggest that IgE synthesis takes place in the lymphoid tissues of Waldeyer's ring and in downstream cervical lymph nodes and that migratory mast cells transport specific IgE back to the nasal mucosa to mediate the allergic reaction. PMID- 3050713 TI - Asymmetry of vertical optokinetic nystagmus and afternystagmus. AB - The sensorimotor mechanism to stabilize the visual field in upright posture is not the same in earth-horizontal plane and vertical plane, because gravity possesses a directionality which is sensed by gravity-receptors. In this communication, asymmetric behavior of the vertical optokinetic nystagmus and afternystagmus is discussed, in conjunction with lesion placements within the peripheral and central neuro-networks (macula sacculi and cerebellar uvula and nodulus) in squirrel monkeys. PMID- 3050718 TI - Rehabilitation of the sports-injured patient. AB - We have presented an overview of basic orthopedic rehabilitative exercises, techniques, and modalities. As our athletic population has become more insistent on performance enhancement techniques, the orthopedic surgeon has adopted a more aggressive approach to rehabilitation of the injured athlete. Many of the rehabilitation techniques and modalities outlined are very inexpensive and require little space; others are very expensive and space-demanding. As our knowledge base expands, we hope we will be able to apply these scientific principles toward better treatment of the injured athlete. PMID- 3050720 TI - Infections of the hand. AB - Important developments in the past 10 years include the use of improved prophylactic antibiotics, anesthetic techniques, jet pulsatile lavage, closed irrigation techniques, postoperative therapy, and splinting. The basic principles of thorough debridement of necrotic and infected tissue and healing by secondary intention with elevation and splinting remain paramount in the treatment of the infected hand. PMID- 3050719 TI - Office evaluation and management of the shoulder impingement syndrome. AB - The shoulder impingement syndrome is the most common cause of shoulder pain. A complete office evaluation can be aided by a printed exam sheet. Careful consideration of the differential diagnosis is important, often employing the Xylocaine injection test. The impingement syndrome responds to conservative treatment the majority of the time. Long follow-up determines the need for an invasive evaluation or surgical treatment. PMID- 3050717 TI - Office evaluation and management of acute orthopedic trauma. AB - Virtually any injury that can be treated as an outpatient can be treated in a properly equipped and staffed orthopaedist's office, provided that it is accurately identified and evaluated initially. Anesthetic blocks, closed manipulations, some suturing, and immobilizations can all be done in the acute setting. Children and elderly patients are particularly suited to office treatment. Close follow-up is essential, and early rehabilitation can begin in the office setting. PMID- 3050721 TI - Evaluation and treatment of congenital dislocation of the hip in infants. AB - This article represents an evaluation and treatment program of CDH in infants. A practical approach describing current trends in diagnostic techniques and treatment modalities are presented for the orthopedist in his office practice. PMID- 3050722 TI - Office evaluation of bone tumors. AB - A familiarity with the characteristics and behavior of musculoskeletal neoplasms will allow for the delivery of timely and appropriate treatment. In this article the significance of historical and clinical data is discussed in general terms, followed by a rationale for office-based evaluation of these lesions. Some of the more common lesions are then discussed in greater detail. PMID- 3050723 TI - A brief history and comparative analysis of disability systems and impairment rating guides. AB - The evaluation of musculoskeletal impairment and disability is an integral part of office practice in orthopedics. Proper performance of this function requires a clear understanding of the evolution and present structure of disability systems and the strengths and weaknesses of impairment rating methodologies. PMID- 3050725 TI - Orthopedic office management. The need for planning and goals. AB - The purpose of this article is to provide physicians in private practice with a methodology for shaping their own destinies and to ensure they will be able to cope successfully with the current changes, as well as those yet to come, that will impact on their practices. It attempts to highlight those core techniques employed in long-term and strategic planning, enabling them to understand what the planning process is all about. PMID- 3050724 TI - Osteoporosis. AB - Osteoporosis is a significant cause of morbidity and mortality in the elderly. As the population in the United States becomes more aged, the magnitude of this problem will certainly become greater. Significant progress has been made in recent years with regard to the diagnosis and treatment of osteoporotic bone disease. Future treatment possibilities being investigated include cyclical coherence programs and cyclical parathyroid hormone administration. While showing some initial promise, these new modalities have been used only in a limited number of patients and must be considered highly experimental. Despite the progress being made in the treatment of osteoporosis, hope for the future clearly rests with prevention. Arresting bone loss has proven to be much more effective than rebuilding a depleted skeleton. Advances must be made in determining the pathogenesis of osteoporosis with the goal of identifying those individuals at highest risk at a young age, when skeletal mass is still increasing. Intervention could then be aimed at maximizing the peak bone density through a combination of proper nutrition, exercise, and lifestyle changes. Clearly, there is much more to learn about this very common and debilitating disease. PMID- 3050726 TI - [Clinical aspects of dermatoplasty in severe open fractures of the extremities]. PMID- 3050727 TI - [Use of mathematical diagnosis for determining the viability of the extremity in open fractures in an experiment]. PMID- 3050728 TI - [Polymer coating of intraosseous fixation devices (experimental research)]. PMID- 3050730 TI - [A substitution method for a defect in the hand bones using a brephograft]. PMID- 3050731 TI - [A fixation device for holding tubular bone grafts]. PMID- 3050732 TI - [Centenary of the birth of Valerian Petrovich Zakharzhevskii]. PMID- 3050729 TI - [Migration of the pin into the popliteal vein after osteosynthesis of the sternoclavicular joint]. PMID- 3050733 TI - [Quantitative assessment of the mineral substances in bone tissue by 2-photon absorptiometry (a review of the literature)]. PMID- 3050734 TI - [Environmental pollution and health preservation]. PMID- 3050735 TI - [Psychic and somatic complaints in the climacteric and during involution after adnexa-preserving hysterectomy]. PMID- 3050736 TI - [Laboratory and morphologic studies in a child with IIb type hyperlipoproteinemia]. PMID- 3050737 TI - [A new era in research and therapy of urolithiasis]. PMID- 3050738 TI - [Diagnostic and therapeutic use of monoclonal antibodies in acute leukemia]. PMID- 3050740 TI - [The first century of the teaching of chemistry and botany at the University of Pest]. PMID- 3050739 TI - [Synthesis of ectopic human chorionic gonadotropins and its significance in gynecologic and colorectal tumors]. PMID- 3050742 TI - [J. E. Purkinje, a supporter of the Czech national language]. PMID- 3050743 TI - [Contribution to the history of first aid in Hungary and the activities of Geza Kresz]. PMID- 3050741 TI - [In memory of Janos Gall, M. D. (1924-1988)]. PMID- 3050744 TI - ["Csokonai and Young"]. PMID- 3050745 TI - [The importance of hygiene in prevention]. PMID- 3050746 TI - [Follow-up examination of children after scarlet fever (study of an l8-year case load]. PMID- 3050747 TI - [Ocular changes in premature infants with a birth weight of 1000 g or less (with special reference to retinopathia prematurorum)]. PMID- 3050748 TI - [Diuretic-induced pseudo-Bartter syndrome in idiopathic edema]. PMID- 3050749 TI - [Prenatal diagnosis of chorioangioma]. PMID- 3050750 TI - [In memory of Laszlo Blaskovics]. PMID- 3050752 TI - [Changes in the "basic principles of socialist public health"]. PMID- 3050751 TI - [From the medical installations of the Roman legions to the new Margit Hospital]. PMID- 3050753 TI - [Simultaneous occurrence of Sheehan syndrome and Willebrand disease]. PMID- 3050754 TI - [Genetic counseling in hydatidiform mole]. PMID- 3050755 TI - [Inaugural address of Frigyes Koranyi]. PMID- 3050756 TI - [S. Tonegawa, 1987 Nobel Prize laureate in medicine]. PMID- 3050757 TI - [Sewall Wright (1889-1988)]. PMID- 3050758 TI - [On the 150th anniversary of the Hungarian Medical Association]. PMID- 3050759 TI - [Genetic program and environmental imprinting: the critical perinatal period]. PMID- 3050760 TI - [Retroperitoneal fibrosis in relation to three reported cases]. PMID- 3050762 TI - [Possibilities of increasing surgical safety at the turn of the millennium]. PMID- 3050761 TI - [Diagnostic and therapeutic difficulties in abdominal actinomycosis in women wearing IUD's]. PMID- 3050763 TI - [Pregnancy produced by intrafallopian gamete transfer]. PMID- 3050764 TI - [Churg-Strauss syndrome following autoimmune thyroiditis associated with Basedow's disease]. PMID- 3050765 TI - [English roots of the Hungarian-language medical works of Istvan Weszpreemi]. PMID- 3050766 TI - [Otto Kahler and multiple myeloma]. PMID- 3050767 TI - [Motion sickness]. PMID- 3050772 TI - [150th anniversary of the birth of Geza Dulacska]. PMID- 3050771 TI - [The death of Sandor Korosi Csoma]. PMID- 3050769 TI - [The role of colfarit in the prevention of pulmonary embolism the aged]. PMID- 3050770 TI - [Herman Boerhaave (1668-1738)]. PMID- 3050768 TI - [The effect of proximal selective vagotomy on the healing of duodenal ulcer]. PMID- 3050773 TI - [The significance of body measurements in the physical examination of infants and children]. PMID- 3050774 TI - [The effect of vitamin A therapy on the immune function and lipid peroxidation in patients with Sjogren syndrome]. PMID- 3050776 TI - [New trends in reproductive medicine--new ethical dilemmas]. PMID- 3050775 TI - [Erythrocyte protoporphyrin studies in hepato-erythropoietic porphyria]. PMID- 3050778 TI - [Chronic gastritis and duodenitis in primary Sjogren syndrome]. PMID- 3050777 TI - [The value of disaccharide determination, associated with histological studies of the small intestine, in various stages if celiac disease and other malabsorption disorders]. PMID- 3050779 TI - [A case of renal hemangiopericytoma]. PMID- 3050780 TI - [Istvan Apathy (1863-1922)]. PMID- 3050781 TI - [History of the library of the Royal Society of Medicine of Budapest]. PMID- 3050782 TI - [A look into the future. 40 years of Austrian Nursing Federation]. PMID- 3050783 TI - [Functional anatomy of the elbow joint]. AB - An attempt has been made to describe the combined action of the bony elements of the elbow joint, together with the capsular ligaments, as a single unit. Particular emphasis is laid upon the mechanisms that prevent abrupt limitation of movement. The account of the periarticular structures, including the adjacent muscles, is followed by a discussion of the various possible surgical approaches to the joint. Finally, a general survey is given of the kinematics and kinetics of the elbow joint. It is suggested that the distribution of the hyaline articular cartilage and the density of the underlying bone are morphological reflections of the stresses acting upon the joint. It follows that the axial pressure acts upon both the ulnar and the radial components of the elbow; therefore, if the joint is to remain stable, at least one of its distal bony elements must be functionally effective. PMID- 3050784 TI - [Congenital changes in the elbow joint]. AB - Radio-ulnar synostosis is one of the congenital malformations of the elbow joint that can be severely disabling, especially if it is present bilaterally or there is marked hyperpronation. In the case of fixed pronation of 60 degrees or more corrective surgery is definitely indicated to reduce functional impairment. As yet, there is no standard operative procedure for restoration of useful supination/pronation function of the forearm. Derotational osteotomy at the site of synostosis seems to be the best procedure. As the postoperative aim we recommend fixed pronation at 15 degrees -35 degrees for the dominant hand and fixed supination at 15 degrees -30 degrees for the contralateral hand. Major derotational procedures should be performed in several stages to avoid neurovascular complications. PMID- 3050786 TI - [Surgical therapeutic possibilities of the elbow in chronic polyarthritis]. AB - The natural history of rheumatoid arthritis of the elbow often includes impairment of the function of the upper extremity in advanced stages of the disease. Synovectomy performed by a large radial incision is considered a worthwhile procedure for stages 1-3 according to the classification of Larsen et al. Radiosynoviorthesis is possible in stages 0 and 1. In the authors' opinion, resection- and interposition arthroplasty remains the procedure of choice for advanced stages 4 and 5. For elbows with severe instability alloarthroplasty may be considered. The radial head should generally not be resected. Entrapment neuropathy of the ulnar and the posterior interosseus nerves is possible in rheumatoid arthritis patients. The surgical treatment consists in decompression, if necessary with transposition and synovectomy of the elbow joint. PMID- 3050785 TI - [Secondary ligament instabilities in the area of the elbow joint]. AB - Secondary capsular or ligamentous injuries around the elbow joint, which are generally a rare finding, can give rise to medial, lateral, anterior, and posterior instability. Medial instability is especially bothersome during sporting activities, such as javelin throwing and baseball, and requires tightening or replacement of the ulnar collateral ligament. On the lateral side it is particularly the subluxation of the head of the radius that makes surgical tightening of the annular ligament essential. Anterior instability, which would cause the joint to dislocate anteriorly, is extremely rare and does not lead to secondary problems. Posterolateral, chronically recurrent, dislocation of the elbow joint, however, is intolerable for the patient, and many operative stabilizing procedures have been performed, mostly in isolated patients, in attempts at correction. The current method of choice consists in tightening of the dorsoradial capsular pouch in combination with osseous refixation of the slack radial ligament. PMID- 3050787 TI - [Neuro-orthopedic problems in the area of the elbow joint]. AB - Neuromuscular impairment of elbow function directly affects the activities of daily living as well as the ability to work in many patients. A series of typical pathophysiological syndromes occur in the region of the elbow as immediate or late results of local trauma or more distant injuries to the brain, cervical cord, cervicobrachial plexus, or peripheral nerves. The operative management is determined by the pathophysiology of the lesions and the anatomical relationship between the muscles and the neurovascular structures at the elbow. These syndromes and their treatment are reviewed. PMID- 3050788 TI - Pennsylvania Medical Society. 1988-89 membership directory. PMID- 3050789 TI - [Sonographic demonstration of intracranial changes in congenital toxoplasmosis]. PMID- 3050790 TI - [The necessity of detecting neonatal seizure activity using polygraphically supplemented electroencephalography]. PMID- 3050791 TI - [Pneumococcal infections in childhood--meningitis. 2: Discussion of the report on 58 cases and literature review]. PMID- 3050792 TI - Clinical application of radioisotopic measurement of the single renal plasma flow (RPF). PMID- 3050793 TI - Pterygium surgery: lamellar keratoplasty. PMID- 3050794 TI - Misdiagnosis of choroidal melanoma. PMID- 3050795 TI - An appraisal of antibiotic policies for urinary tract infections in patients with spinal cord injuries undergoing long-term intermittent catheterisation. AB - The antibiotic policies that have been recommended in the literature for the prevention and treatment of urinary tract infections in patients with spinal cord injuries undergoing long-term intermittent catheterisation are reviewed. Current practices in spinal units in England and Wales are reported and the rational bases of these policies are considered. PMID- 3050796 TI - Haemodynamic, hormonal and urinary responses to postural change in tetraplegic and paraplegic man. AB - We have studied the haemodynamic, hormonal and urinary effects of postural change in 6 tetraplegic patients, 6 paraplegic patients and 6 normal subjects. Measurements of blood pressure and heart rate, plasma renin activity, plasma aldosterone, urine volume and electrolyte excretion were made for 60 minutes while sitting and 60 minutes while recumbent. In tetraplegics the blood pressure was lower when sitting and rose during recumbency, unlike paraplegics and normal subjects. Plasma renin activity and aldosterone were higher in tetraplegics when sitting compared to normal subjects and did not fall during recumbency. Urine output increased significantly after recumbency in tetraplegics, but not in paraplegics or normal subjects. Both urinary sodium and potassium excretion were lower in tetraplegics and higher in paraplegics compared to normal subjects when sitting. In paraplegics the fall in both sodium and potassium excretion did not appear to be related to change in posture. Our observations indicate that recumbency induces a diuresis in tetraplegics but not in paraplegics or in normal subjects. The diuresis in tetraplegics may be related to the accompanying haemodynamic and hormonal changes induced by recumbency. PMID- 3050797 TI - Non-traumatic ischaemic myelopathy: a review of 25 cases. AB - The causes of ischaemic myelopathy are described in 25 patients. Nine developed following surgical manipulation or traumatic laceration of the aorta, 1 following intercostal artery ligation, 3 following aortic aneurysm dissection, 2 following myocardial infarction and/or cardiac arrest, 7 in the absence of any specifically identifiable predisposing factors, and 3 in association with decompression sickness. The degree of clinical recovery was greater among those with incomplete spastic (as opposed to complete flaccid) paralysis and among those in whom sensory loss below the level of injury was incomplete. Despite the diversity of mechanisms that may lead to the development of spinal cord ischaemia, structural damage seems in most instances to affect either grey matter or white matter predominantly. Some of the possible reasons for these preferential sites of damage are discussed. PMID- 3050798 TI - Joe Bousquet: paraplegia as a poet's plight and challenge. PMID- 3050799 TI - Macrophage activation in vitro by lymphocytes from Leishmania major infected healer and non-healer mice. AB - Peritoneal macrophages from CBA/T6 (healer) and BALB/c (non-healer) mice were infected with Leishmania major (LV39) in vitro. The microorganism replicated at the same rate in macrophages from either strain. Exposure of infected cells to lymph node cells (LNC) from infected syngeneic animals led to intracellular killing of the parasite by macrophages from both strains, provided LPS was present in the incubation medium. In vitro-propagated L.major-specific T-cell blasts activated macrophages from either strain in the absence of LPS. On a per cell basis, lymphoid cells from BALB/c mice were less efficient, however, than cells from CBA/T6 mice. Lysis of parasitized macrophages was also more marked in CBA/T6 than in BALB/c cell mixtures. LNC exposed to parasite antigen or to infected macrophages secreted macrophage-activating factor (MAF); incubation with antigen also induced lymphocyte proliferation. MAF production and LNC proliferation decreased with progression of the infection of BALB/c mice, but always remained significant. The reduction in relative T-cell numbers in the lymph nodes of infected animals was moderate; the absolute number of T-cells increased markedly in the lymphoid organs of both strains, however. These results suggest that failure to heal may coexist together with active cell-mediated immune response in non-healer mice. PMID- 3050801 TI - Primary nursing in a pain center. PMID- 3050800 TI - Reactivity of stage-specific monoclonal antibody 1G7 with metacyclic trypomastigotes of Trypanosoma cruzi strains: lytic property and 90,000 mol. wt surface antigen polymorphism. AB - Eleven strains of Trypanosoma cruzi, originating from a variety of vertebrate and invertebrate hosts in distinct geographical regions, were examined for the reactivity of metacyclic stages with the monoclonal antibody 1G7. Trypomastigotes of five strains were susceptible to complement-dependent 1G7-mediated lysis. Higher levels of 1G7 bound to metacyclics of lysis-susceptible strains as compared to lysis-resistant isolates. Excluding Y and CL strains, 1G7 reacted with metacyclics of all T. cruzi isolates by binding to a 90,000 mol. wt surface polypeptide. A 90,000 mol. wt protein lacking the 1G7-specific epitope but immunologically related to the 90,000 mol. wt antigen of other T. cruzi isolates is present in Y and CL strains. The intensity of the 90,000 mol. wt band, detected by surface iodination of metacyclics or in immunoblots using the monoclonal antibody 1G7 or the monospecific antiserum to 90,000 mol. wt protein, varied among different strains and also a discrete variation was observed in its molecular weight. The overall analysis reveals a polymorphism of the 90,000 mol. wt protein, which is ubiquitous among the different T. cruzi isolates. PMID- 3050802 TI - Computed tomography of the lower extremity. Part II. PMID- 3050803 TI - Osteonecrosis of the femoral head. PMID- 3050804 TI - Pigmented villonodular synovitis presenting as a large lateral knee mass. Case report and review of the literature. AB - Pigmented villonodular synovitis is characterized by synovial villonodular lesions with hemosiderin pigmentation and a stromal infiltrate of histiocytes. Analogous lesions may occur in joints, bursae, and tendon sheaths. Although the etiology and natural history of pigmented villonodular synovitis remain unknown, it should be considered a benign entity. It usually presents as either diffuse monoarticular involvement with chronic pain and swelling, or a more localized nodular lesion, typically involving the fingers or knees. Although treatment involves excision of the involved tissue, due to its benign behavior, radical surgical procedures are not indicated. A case report of a large, extra-articular mass is described, with histologic and clinical changes consistent with localized PVS. PMID- 3050805 TI - Total dislocation of the talus. Case report with a 13-year follow up and review of the literature. AB - Simultaneous dislocation of the talus from the tibiotalar, talocalcaneal, and talonavicular joints is uncommon and usually occurs from considerable violence. Total dislocation of the talus is frequently an open injury, or the skin may be tented over the dislocated talus leading to skin slough. Infection and avascular necrosis may follow. Treatment options include closed or open reduction, talectomy, and arthrodesis. The following case and literature review illustrate some of the problems encountered and factors that influence the outcome of this injury. PMID- 3050807 TI - Computed tomography of the lower extremity. Part III. PMID- 3050806 TI - Fractures of the scapula. A review of the literature. AB - Scapular fractures, a relatively uncommon injury, most often result from major trauma. This paper categorizes the different types of scapular fractures; outlining clinical presentation, diagnosis, treatment, and complications. The importance of a good physical examination is stressed; as overlooking a soft tissue injury (i.e., pneumothorax, neurovascular tear, cerebral contusion) commonly associated with scapular fractures may be life-threatening. Scapular fractures are commonly treated nonoperatively. However, when glenohumeral joint function is impaired, surgery may be indicated after patient age, occupation, and clinical status are evaluated. PMID- 3050809 TI - Hip pain in children: an anatomic approach. AB - The practicing physician can be assured of both an accurate and systematic method for making a diagnosis when using an anatomic approach to the common clinical problem of hip pain. The numerous causes of hip and leg pain in the small child and adolescent and their management are reviewed. PMID- 3050810 TI - Arthroscopic surgery of the ankle. AB - Ankle arthroscopy is a useful technique, especially for intra-articular problems and osteocartilagenous lesions. Indications for this diagnostic and therapeutic technique include: ankle sprain unresponsive to the usual course of treatment, complaints of intermittent locking, pain and clicking, limitation of motion, and painful conditions with no obvious etiology (especially in a worker's compensation beneficiary). A review of arthroscopic, radiographic, and clinical data of all patients undergoing ankle arthroscopy at our center provided the following diagnoses: talar dome osteochondral fractures, loose bodies, accessory ossicles, talar dome cyst with loose bodies, and chronic synovitis. The anatomy of ankle arthroscopy is demanding; while the technique incorporates the same instrumentation as is used for knee arthroscopy, awareness of the neurovascular structures is essential to avoid complications. Although infrequent, complications can include sinus tract formation, sensory nerve damage, synovitis, infection, instrument breakage, and calf compartment syndromes due to extravasation of irrigation fluid. PMID- 3050808 TI - Computed tomography of the lower extremity. Part IV. PMID- 3050811 TI - Pediatric update #4. Familial congenital short femur: intrauterine detection and follow-up by ultrasound. A case report. AB - A case of familial congenital simple short femur is described in an 8-year-old brother and an 8-month-old sister, associated with other congenital defects in the lower extremities. In the case of the sister, early intrauterine detection and follow-up were possible. This early intrauterine detection by means of ultrasound may enable us to calculate the ratio between the length of the short femur and that of the normal one and to plan for corrective surgery at a later date. PMID- 3050812 TI - Trauma and the orthopaedic surgeon. PMID- 3050813 TI - Computed tomography of rheumatologic disorders. Part II. AB - The advent of computed tomographic (CT) scanning has initiated a revolutionary approach to the evaluation of articular disease. CT affords the opportunity to noninvasively evaluate both osseous and soft tissue structures during a single diagnostic examination. An optimal CT study demands proper technique, including a localization image, appropriate slice thickness and spacing, and window/level manipulation. The examination must be tailored to the specific indication for the study, which may include degenerative processes, inflammatory disease, or unexplained clinical symptomatology. PMID- 3050814 TI - Complex dislocations of the metacarpophalangeal joints. AB - Complex dislocations of the metacarpophalangeal joints, those blocked by soft tissue interposition, occur infrequently and are often misdiagnosed and mishandled. While these injuries can be treated in a closed fashion, they may actually be induced by incorrect technique in reducing simple MP dislocations. Open surgical reduction is the treatment of necessity, although there is currently no consensus on approach. This article presents an overview of the history, pathoanatomy, and clinical and radiographic findings of this fairly uncommon hand injury. The literature on the various treatment modalities is reviewed, and therapeutic guidelines are offered. PMID- 3050815 TI - Asymptomatic innominate vein tamponade with retromanubrial clavicular dislocation. A case report. AB - Traumatic sternoclavicular joint dislocations are uncommon and posterior dislocations are rare; however, reports in the orthopaedic literature focus predominantly on retrosternal dislocations owing to their potentially dangerous sequelae. We report a case of asymptomatic complete innominate vein obstruction secondary to retromanubrial dislocation of the sternoclavicular joint with restoration of flow after closed reduction. PMID- 3050817 TI - Approaches to senior care #3. Orthopaedic management of the stroke patient. Part II: Treating deformities of the upper and lower extremities. AB - Cerebrovascular accidents are among the most serious medical problems in the United States. In Part I of this report, the pathophysiology, types of impairment, and evaluation of the stroke patient were discussed. In this report, Part II, the management of extremity deformities will be reviewed. PMID- 3050816 TI - Bilateral simultaneous infrapatellar tendon rupture: support for Davidsson's theory. AB - Rupture of the extensor mechanism of the knee is not an unusual occurrence. Bilateral simultaneous rupture, however, is rare. Most cases of bilateral simultaneous rupture occur in association with systemic disease, i.e., systemic lupus erythematosus, arteriosclerosis, diabetes, or secondary hyperparathyroidism. Only three cases without predisposing conditions have been reported. A case report of a 48-year-old black male without apparent risk factors who sustained spontaneous simultaneous rupture of the patella tendons is presented. Histologic evidence supports Davidsson's theory of multiple recurrent microtears within the tendon substance as a cause of rupture. PMID- 3050818 TI - Immunocytochemical demonstration of a neuropeptide in Ascaris suum (Nematoda) using an antiserum to FMRFamide. AB - A FMRFamide-like peptide has been detected in the nematode Ascaris suum, using the peroxidase-anti-peroxidase (PAP) immunocytochemical technique. Positive reactions were obtained in both the central nervous system and the peripheral nervous system of the worm, the strongest reactions being in the anterior nerve ring, the cephalic papillary ganglia, the lateral ganglia and the dorso-rectal ganglion. Immunoreactivity was observed along the length of the main nerve cords of the worm and, to a lesser extent, in the pharyngeal nerve cords. The possible role of this neuropeptide in the physiology of the nematode is discussed. PMID- 3050819 TI - [Residual protease activity of the gastric mucosa after histamine stimulation normally and in acetate-induced ulcer in arts]. PMID- 3050820 TI - [Oxygen supply of the body during the administration organofluorine emulsions (a pathophysiological analysis)]. PMID- 3050821 TI - Assessing Pavlov's impact on the American conditioning enterprise. AB - In this article, the visibility of Pavlov and of Watson in American psychology are compared, and the periods of their respective influence are specified with greater precision than is afforded by merely impressionistic methods. The author also critically examines the possibility that the early history of the American classic-conditioning enterprise involved a succession of two phases: a Watsonian/speculative phase and a Pavlovian/empirical phase. In conclusion, the author assesses the possibility that the publication of Pavlov's Conditioned Reflexes (1927) "stimulated" scholarly work on Pavlovian conditioning, and finds this proposition lacking empirical support. PMID- 3050822 TI - Pavlov and the Rockefeller Foundation. AB - Despite the tension between the United States and the Soviet Union in the early 1920's, the Rockefeller Foundation and the Rockefeller Institute for Medical Research found ways to assist I.P. Pavlov. In addition to providing scientific literature and financial aid, these institutions and their officers rendered important moral support to the scientific career of Pavlov during his later years. In 1923, as a guest of the Rockefeller Institute, Pavlov visited American scientific laboratories. In 1924, he requested and received a number of books on physiology, and during the 1930's the Foundation helped him to acquire equipment for his Leningrad laboratory. PMID- 3050823 TI - Imitative behavior. A theoretical view. AB - This article reviews the results of experimental studies on imitative behavior reported by various investigators, and then discusses the possible brain mechanisms responsible for this behavior. It was found that human infants in their first hours of life were already capable of spontaneous imitation of simple motor acts demonstrated by an adult, without previous training or reward; these observations suggest that imitative behavior is an innate process that can be considered an unconditional reflex of imitation. It was also found that satiated animals resumed eating when they saw their companions eating. In the latter case, the imitative reflex triggered the previously acquired feeding behavior. Similar mechanisms could be responsible for the phenomenon of eating more in the presence of companions than in their absence, as well as that of preferring the food chosen by companions. When followed by a reward, the imitative act can be learned -that is, transformed into an instrumental conditional response; learning by imitation of simple motor acts was observed in animals, and that of complex motor acts was observed in children who had already achieved a certain developmental stage. In animals, learning complex motor tasks was facilitated by previous observation of a companion performing this task. In this case, the presence of the observer during the session could lead to habituation of the experimental situation and production of associations between this situation and stimuli or emotions related to the reward or punishment, and might result in more efficient learning later. The imitative behavior can be inhibited by stimuli producing responses antagonistic to the act of imitation. PMID- 3050824 TI - Movement as a signal during classical conditioning. AB - Analysis of the available data and that of the author disclosed the peculiarities of motor reaction when used as a conditioned stimulus. The author's data showed that if signal value is attributed to a motor reaction (passive movement or movement evoked by the direct stimulation of the motor cortex), the changes of excitability in the motor cortex representation of the dog's leg depend on the biological sign of the reinforcing stimulus during classic conditioning. They also remained the same during instrumental conditioning and were opposite in sign, showed increased excitability in the food situation, and decreased excitability in the defense situation. Using the movement as a conditional stimulus, we managed to uncover the commonality between classic and instrumental conditioning. This enabled us to answer questions, discussed by Pavlov and Guthrie, which, it seems to us, had not been convincingly answered during their time. PMID- 3050826 TI - A practical guide to high-frequency ventilation. PMID- 3050825 TI - Surfactant therapy in the newborn. PMID- 3050827 TI - Extracorporeal membrane oxygenator therapy. PMID- 3050828 TI - The management of hypoxic-ischemic encephalopathy. PMID- 3050829 TI - Retinopathy of prematurity: progress report. AB - For the time being, risk of ROP seems inexorably linked to survival of extremely low birth weight prematures, whose embryonic retinas develop in an abnormal and fluctuating environment. Incidence and severity may be reduced by stabilizing ventilation, oxygenation, and perfusion, moderating light exposure, and providing normal levels of vitamin E. Progression of stage 3 retinopathy may sometimes be arrested by cryotherapy. In the future, antioxidants or other pharmacologic agents may be developed to provide a greater margin of safety. In the meantime, the eyes of prematures must be examined and those with ROP will need specialized ophthalmologic care. PMID- 3050830 TI - The wheezing infant. AB - In summary, wheezing is a common manifestation of viral respiratory tract disease in infancy. The precise pathogenetic mechanisms of virus-induced wheezing and its sequelae are not clear, although recent reports about participation of the cellular and humoral immune systems are promising. Although therapies like those used to treat asthma are employed in the treatment of virus-induced wheezing in infancy, their efficacy remains controversial in bronchiolitis. Recently developed agents with antiviral properties are promising and the choice of any of these agents in a therapeutic regimen should be individualized. Antiviral agents during acute infections may modify the long-term sequelae. Clearly, much work needs to be done to elucidate pathogenetic mechanisms, and to develop new, safe, and effective anti-inflammatory agents for the therapy of these disorders. PMID- 3050831 TI - Exercise-induced bronchospasm in children and adolescents. AB - The early recognition and appropriate management of EIB can allow children and adolescents to participate fully in physical activities and sport. The diagnosis by history of chest congestion, coughing, and decreasing performance with exercise is helpful but is aided by a more systematic questionnaire that can detect otherwise "normal" people with EIB. The diagnosis is documented by performance of an exercise challenge test such as a treadmill or cycloergometer to verify bronchospasm induced by exercise. The management can be accomplished by nonpharmacologic means such as an early vigorous warm-up, the use of a mask for rebreathing warmed air, and participation in a physical training program to increase anaerobic fitness. Pharmacologic management includes the appropriate use of cromolyn sodium, beta-adrenergic agonists, theophylline, ipratromium bromide, and calcium channel blocking agents. In addition, the antihistamine, terfenadine, can be used to block EIB effectively. These pharmacologic agents can be utilized in both national and international competition when approved by the appropriate national governing body or the U.S. Olympic Committee and the International Olympic Committee. PMID- 3050832 TI - Asthma: current therapeutic approach. AB - This article discusses the goals of asthma therapy, treatment of the acute exacerbation, and day-to-day management. The importance of early pharmacologic intervention is emphasized. In 1988, the need to hospitalize a child with asthma often represents failure of ambulatory management. PMID- 3050833 TI - Otitis media and its relationship to allergy. AB - The importance of infection in the etiology of otitis media and the role of eustachian tube obstruction in the pathogenesis of otitis media with effusion are well known. Recently, allergic rhinitis has been documented to induce eustachian tube obstruction. When allergic rhinitis is diagnosed in a child with recurrent or chronic middle ear disease, allergy should be considered as another risk factor for the development of otitis media with effusion. PMID- 3050834 TI - Chronic sinusitis in the allergic child. AB - Chronic inflammation of the paranasal sinuses, especially the maxillary sinuses, is common in children with respiratory allergy. The presence of sinusitis should be suspected in such children when they have chronic night and early morning cough or poorly controlled asthma. Treatment should be aggressive because morbidity can be high. PMID- 3050835 TI - Immunotherapy for allergic respiratory disease. AB - In summary, immunotherapy is probably a useful treatment for selected cases of allergic airway disease. Patients should be selected who are allergic and who have at least moderately severe illnesses. Using commercially available solutions of antigen extracts, a therapeutic response may be expected in about three quarters of patients in that they will be able to reduce or eliminate medications. The disadvantages of treatment include its inconvenience and cost, its frequent ineffectiveness, and its risks of anaphylaxis; many of these disadvantages are eliminated by newer antigen preparations. PMID- 3050836 TI - Food hypersensitivity and atopic dermatitis. AB - Initially, this article focuses on the pathogenesis of IgE-dependent immediate and late-phase responses in pediatric patients with atopic dermatitis. The article also discusses the role of food hypersensitivity as a major trigger factor exacerbating atopic dermatitis in children. Finally, consideration is given to the prevention of atopic disease through exclusive breast feeding. PMID- 3050837 TI - IgE response and its regulation in allergic diseases. AB - The distinguishing feature of the allergic person is his or her elevation of serum IgE. This propensity to develop a sustained IgE response is determined genetically. The biologic effects of IgE are mediated via Fc receptors (Fc epsilon R) present on mast cells and basophils (Fc epsilon R type 1) and subpopulations of monocytes, macrophages, eosinophils, and platelets (Fc epsilon R type 2). Interaction of allergen with IgE on these cells results in receptor "bridging" and the release of histamine and other inflammatory mediators. Fc epsilon R type 2 on lymphocytes and monocytes are upregulated in atopic disease and may play a role in the allergic inflammatory reaction. The activation of B cells to synthesize IgE requires several stages (see Fig. 2). T cells play an important role in the regulation of IgE synthesis. In vitro activation of resting B cells to synthesize IgE requires direct cellular interaction with T cells or the presence of IL4 for activation. The latter effect is inhibited by alpha interferon. Preactivated B cells are influenced in an isotype-specific manner by T-cell-derived IgE binding factors (IgE-BF), which may act as IgE-potentiating or IgE-suppressive factors, depending on their degree of glycosylation. The regulation of IgE synthesis is an important area of investigation. It provides us with an understanding of the basis of the human allergic response and ultimately may provide the basis for novel strategies in the treatment of allergic diseases. PMID- 3050838 TI - Early/late response model: implications for control of asthma and chronic cough in children. AB - The observed airways inflammation in asthma and chronic cough supports the conclusions of clinical trials, namely, that our treatment regimens should emphasize inhaled cromolyn. The need for bronchodilators as backup therapy is real but represents testimony to the fact that in some cases it has not been possible to entirely eliminate the inflammation and the consequent airways hyperresponsiveness. PMID- 3050839 TI - Allergens: recent advances. AB - This article summarizes the recent advances in allergen isolation, characterization, standardization, and nomenclature. Newer methods of quantifying amorphous aeroallergens also are reviewed. PMID- 3050840 TI - Allergy testing: in vivo versus in vitro. AB - Table 4 briefly summarizes the relative advantages and disadvantages of skin tests versus in vitro tests for detecting allergen-specific IgE. The skin test remains unexcelled as a sensitive and cost efficient test for specific IgE. The high degree of skin test sensitivity is very important when a patient must be evaluated for potentially life-threatening allergies such as to penicillin or stinging insects. The results of both skin tests and in vitro assays depend very much on the quality of the allergen extracts used for the tests. Although the quality of extracts is improving, there is still little standardization. Both skin tests and in vitro assays are difficult to quality control. Practicing allergists rely on experience, and the correlation between patient histories and skin tests results for quality control of the results. Although this system suffices for common allergens, the results for uncommon allergens may be misleading. Quality control is also difficult for in vitro tests. Participation in quality control programs, such as that being offered by the College of American Pathologists, will increase and lead to better quality and standardization of in vitro test results. At the present time, properly performed skin tests are the best available method for detecting the presence of allergen specific IgE. They are rapid, sensitive, and inexpensive on a per test basis. In vitro tests are acceptable substitutes for skin tests in some circumstances. If the patient does not have normal skin, cannot discontinue interfering medications, or is so sensitive by history that anaphylaxis seems possible, in vitro tests are preferred. In vitro tests are better when it is necessary to test a difficult patient such as a combative, mentally retarded adult. In vitro tests also have been invaluable in many allergy research studies. Physicians must remember that positive tests for allergen-specific IgE do not diagnose allergy. They only indicate the presence of IgE molecules with a particular immunologic specificity. A decision whether the specific IgE molecules are responsible for clinically apparent disease must be made by a well-trained physician. The ultimate standard for the diagnosis of allergic disease remains the combination of: (1) positive double-blind challenge, (2) the presence of specific IgE, and (3) demonstration that the symptoms are the result of IgE-mediated inflammation. PMID- 3050841 TI - Primary ciliary dyskinesia: evaluation and management. PMID- 3050842 TI - Solitary myeloblastoma presenting as acute hydrocephalus: review of literature, implications for therapy. AB - An unusual presentation of a case of myeloblastoma (granulocystic sarcoma, chloroma) as demonstrated by computed tomography (CT) and selective angiography is reported. Our patient, who presented with acute hydrocephalus due to a large posterior fossa myeloblastoma, had no evidence of systemic disease in either peripheral blood smear, bone marrow aspiration, lumbar puncture or further metastatic workup. While no evidence for generalized disease was seen, systemic chemotherapy may help to prevent overt systemic leukemia. PMID- 3050843 TI - Acute renal infarction: diagnosis by Doppler ultrasound. AB - A young infant undergoing balloon angioplasty for pulmonic stenosis developed global right renal infarction as a complication of cardiac catheterization via the femoral vein. A second patient developed segmental infarction of a recently transplanted kidney due to occlusion of a polar artery. Although routine renal sonography of these patients was normal, duplex Doppler sonography documented the absence of renal arterial and venous flow. PMID- 3050844 TI - Sonographic detection of occult bone fractures. AB - Two infants of 24 and 20 months of age with painful local swelling at the femoral and clavicular regions were investigated by ultrasound after a negative radiographic study of the adjacent bones. In both children high resolution ultrasound clearly revealed the presence of bone fractures in addition to the soft tissue hematomas. These fractures were confirmed by a repeat radiographs performed 6 and 8 days later. Although sonography is not the method of choice for the detection of bone fractures, it may be worthwhile to examine the bone contour for a fracture when a painful swelling adjacent to bone is present. The method may be particularly rewarding in children due to its rapid non-invasive nature and to the small tissue thickness that has to be penetrated. PMID- 3050845 TI - Cystic neuroblastoma with colonic fistula. AB - Neuroblastoma is the most common malignant tumour in infancy originating in about 70% of cases in the adrenal gland. Haemorrhage and necrosis is often seen in neuroblastoma but cyst formation is uncommon. Fistulous communication between an adrenal cystic neuroblastoma and the large bowel has never to our knowledge been reported before. PMID- 3050846 TI - Multiple subdural abscesses following colonic perforation--a rare complication of a ventriculoperitoneal shunt. AB - A case of colonic perforation by a ventriculoperitoneal shunt, its subsequent migration and protrusion from the anal orifice is reported. The shunt reservoir and ventricular catheter were removed percutaneously, and the disconnected peritoneal catheter was pulled out through the anus. Blood culture grew Klebsiella pneumoniae and Streptococcus fecalis. CT scan showed multiple subdural abscesses with evidence of ventriculitis. Removal of the shunt (as described), evacuation of subdural pus and systemic antibiotics resulted in complete recovery. PMID- 3050848 TI - Acute acalculous cholecystitis in salmonella infection. PMID- 3050847 TI - Lipoma of the corpus callosum: diagnosis by ultrasound and magnetic resonance. AB - A lipoma of the corpus callosum in an infant is evaluated by ultrasound and magnetic resonance. The appearances are correlated as a diffuse but continuous aspect of this midline anomaly. PMID- 3050849 TI - Ultrasound diagnosis of adrenal hemorrhage in meningococcemia. AB - Two cases of adrenal hemorrhage in meningococcemia detected by ultrasound are reported. Antemortem detection of adrenal pathology may have important prognostic and therapeutic implications. The sonographic appearance may be echo-free, mixed, or echogenic. Abdominal ultrasound examination can be an effective non-invasive tool in diagnosing adrenal hemorrhage. PMID- 3050850 TI - Effects of captopril on the distribution of left ventricular output with ventricular septal defect. AB - The renin-angiotensin system is activated by congestive heart failure associated with a ventricular septal defect (VSD). To determine the effect of angiotensin converting enzyme inhibition on the hemodynamics with VSD, the dose response curve of captopril was measured in lambs. Furthermore, the effect of captopril on the distribution of systemic output was determined by the radionuclide-labeled microsphere technique. A total of 12 lambs (less than 1 month old) with VSD were instrumented and a minimum of five animals was tested for each data point. Captopril (0.05-10 mg/kg) caused dose-dependent vascular changes. At a dose of 2 mg/kg, maximal hemodynamic effects were observed. The systemic resistance fell by 28 +/- 9% (mean +/- SD, p less than 0.05, n = 9), whereas pulmonary arteriolar resistance rose by 113 +/- 34% (p less than 0.05). These vascular changes caused a reduction in the ratio of pulmonary to systemic flow from 3.31 +/- 0.59 to 2.19 +/- 0.29 (-34%, p less than 0.05) and a reduction in left-to-right shunt volume by 30% (p less than 0.05). The left atrial pressure fell from 17.3 +/- 3.4 to 10.8 +/- 2.8 mm Hg (-38%, p less than 0.05). Mean aortic pressure was unchanged (71.2 +/- 8.1 versus 67.4 +/- 9.1). Forward flow from the left ventricle increased from 2.17 +/- 0.46 to 2.86 +/- 0.54 liter/min/M2 (p less than 0.05). Microsphere-determined organ blood flow to the heart, kidney, liver, duodenum, and skeletal muscle was preserved after a 5 mg/kg dose of captopril and, in fact, tended to increase, but the changes were not significant.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3050851 TI - Howland Award presentation to Joseph Dancis. PMID- 3050852 TI - Does Ureaplasma urealyticum cause respiratory disease in newborns? PMID- 3050853 TI - Immunogenicity and reactogenicity of four Haemophilus influenzae type b capsular polysaccharide vaccines in Finnish 24-month-old children. AB - Stimulated by questions raised on potential differences between the Haemophilus influenzae type b capsular polysaccharide (PRP) vaccines on the market and the applicability of the efficacy data of a similar PRP vaccine obtained in a large field study in 1974 in Finland, we wished to compare the immunogenicity of all these preparations in 24-month-old Finnish children. In our study 137 children received the now recommended 25-micrograms dose of 1 of 4 H. influenzae type b PRP vaccines currently available, and an additional 86 children received half this dose corresponding to the 12.7 micrograms used in 1974. Anti-PRP antibodies were measured in blood samples taken before and 4 weeks after vaccination by the same radioimmunoassay and in the same laboratory as in 1974. The vaccines were equally immunogenic. Furthermore the now recommended dose of 25 micrograms did not give results (geometric mean concentrations, 2.38 to 3.45 micrograms/ml) differing from those after a 12.5-micrograms (2.01- to 3.45-micrograms/ml) dose which was used in 1974. Antibody concentrations of 0.15 and 1.0 micrograms/ml were achieved in 91 to 95 and 66 to 84% of the children, respectively. The corresponding values after 1974 vaccinations were 3.53 micrograms/ml and 100 and 82% of children, respectively. The percentage of those responding with concentrations greater than or equal to 1 microgram/ml was somewhat higher in these Finnish children than reported for children of the same age receiving the same vaccine lots in the United States. Adverse reactions were mild or moderate and transient. PMID- 3050855 TI - Childhood immunization, vaccine-preventable diseases and infection with human immunodeficiency virus. PMID- 3050854 TI - Group A streptococcal infection in children younger than three years of age. AB - We evaluated 758 sick children younger than 3 years of age for Group A beta hemolytic streptococcal (GABHS) upper respiratory infection (URI) to determine the usual clinical presentation of the disease in this age group, indications for culture and the optimal site(s) from which to isolate the organism. GABHS infection was documented in 35 subjects (4.6%). The classic presentation (as proposed in the 1940s) of GABHS URI in children younger than 3 years of age was not confirmed by this study. In 32 of the GABHS cases there were pharyngitis, common cold symptoms or both, and these were associated with acute otitis media 10 times and with otitis media with effusion 3 times. Clinical impetigo was associated with GABHS URI (4 of 32 cases). GABHS URI would not have been documented in 6 of 32 cases if cultures of the anterior nares had not been performed. Children between 18 and 36 months of age were more likely to have GABHS disease than were younger children. Hoarseness and vomiting occurred less frequently in children younger than 36 months with GABHS infection than in those of that age who had non-beta-hemolytic streptococcal illnesses. A history of two or more siblings at home or a family member with a recent streptococcal infection and the presence of irritability, a reddened throat or palate or uvular edema were each associated with GABHS URI. We concluded that sick children between 18 and 36 months of age with a reddened throat should have cultures taken of the throat and anterior nares for GABHS.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3050857 TI - Consensus: management of Salmonella infection in the first year of life. PMID- 3050856 TI - Treatment of severe typhoid fever in children with high dose dexamethasone. PMID- 3050858 TI - Mortality and morbidity from invasive bacterial infections during a clinical trial of acellular pertussis vaccines in Sweden. AB - A double blind placebo-controlled efficacy trial of two acellular pertussis vaccines was conducted in 3801 6- to 11-month-old children. Four vaccinated children died during 7 to 9 months follow-up as a result of Haemophilus influenzae type b meningitis, heroin intoxication with concomitant pneumonia, suspected septicemia, and Neisseria meningitidis Group B septicemia. From the actual death rate in children belonging to the same birth cohort in Sweden that could have been eligible for the trial, one death was expected among vaccinated children. Several investigations were carried out to examine the possibility that the deaths could be causally related to the vaccination. The relative risk for hospitalization due to systemic or respiratory infections was 1.07 (95% confidence interval, 0.95 to 1.20) and 0.83 (95% confidence interval, 0.64 to 1.08) in the vaccine groups as compared with the placebo group. Subsets of the population were studied for signs of immunosuppression. There was no indication of immunoglobulin deficiency or any sign of clinically significant leukopenia or lymphocytosis in vaccine recipients. The results of this analysis provide no evidence for a causal relation between vaccination with the studied acellular pertussis vaccines and altered resistance to invasive disease caused by encapsulated bacteria. The hypothesis that the two variables are related, however, cannot be refuted from these data. PMID- 3050859 TI - Childhood infection caused by Ehrlichia canis or a closely related organism. PMID- 3050860 TI - Failure of direct fluorescent antibody staining to detect Chlamydia trachomatis from genital tract sites of prepubertal children at risk for sexual abuse. PMID- 3050861 TI - Albendazole treatment of recurrent echinococcosis. PMID- 3050862 TI - Mixed flora brain abscess with Pseudomonas paucimobilis after a penetrating lawn dart injury. PMID- 3050863 TI - Chlamydia and sexual abuse. PMID- 3050865 TI - Rapid group A streptococcal antigen detection kit: effect on antimicrobial therapy for acute pharyngitis. AB - Newly introduced rapid diagnostic tests for group A streptococcal pharyngitis should facilitate appropriate antimicrobial use in patients with group A streptococcal pharyngitis. Because of high rates of acute pharyngitis in Tuba City, AZ, at the Navajo Indian reservation, the use of rapid diagnostic test was prospectively evaluated. The sensitivity and specificity of the test was measured and changes in physician prescribing patterns attributable to use of the test were correlated. Of 320 patients with pharyngitis enrolled during the present 3 week study, 86 met the study's definition of a patient with streptococcal pharyngitis and 163 met the study's definition of a patient with nonstreptococcal pharyngitis. The rapid test was 62.8% sensitive and 96.9% specific in identifying patients from whom group A streptococci were isolated. Although treatment of patients with streptococcal pharyngitis at the time of the first visit increased from 36.5% in a retrospective sample to 72.5% during the study, treatment of patients in whom cultures were negative remained the same. Further analyses showed that physicians tended to treat patients with signs characteristic of streptococcal pharyngitis and, as the study progressed, to rely less on negative rapid test results as a reason to withhold antimicrobial agents. It was concluded that rapid tests with good specificity but limited sensitivity may improve treatment of patients with streptococcal pharyngitis by allowing earlier specific therapy. A more sensitive test with a higher negative predictive value would be necessary to prevent treatment of persons with nonstreptococcal pharyngitis. PMID- 3050864 TI - Hemorrhage, phenobarbital, and fluctuating cerebral blood flow velocity in the neonate. AB - Fifty-one sequential intubated babies with birth weights of less than 1,751 were evaluated by serial Doppler ultrasound during the first three days of life. These babies were part of a phenobarbital prophylaxis trial cohort study. Subependymal intraventricular hemorrhage developed in 17 of the babies. Infants with subependymal-intraventricular hemorrhage, whether or not they received pancuronium or phenobarbital, had coefficients of variation comparable to those of babies without hemorrhage. Coefficient of variation values of the right were comparable to values obtained from the left anterior cerebral artery complex and did not appear to be consistently altered by the presence of subependymal intraventricular hemorrhage. Coefficient of variation values appeared to be consistently greatest on day 1 and lowest on day 2. In addition, the values overall increased as the number of waves used to determine the coefficient of variation enlarged from five to 20. This phenomena, however, was not seen among pancuronium recipients and suggests that movement artifact may be a determinant of coefficient of variation values. We conclude that, when the best 20 waves are chosen to evaluate the coefficient of variation, no association exists between coefficient of variation values and development of subependymal-intraventricular hemorrhage or administration of phenobarbital. PMID- 3050866 TI - Ultrasound screening of healthy infants for urinary tract abnormalities. AB - The purpose of the study was to determine the incidence of silent but significant urinary tract abnormalities that might be detected by screening renal ultrasound studies of apparently healthy infants. Of 437 babies studied, six were found to have uropathology severe enough to warrant surgery for an incidence of 1.37% or one of every 73 babies studied. PMID- 3050867 TI - Decreased neonatal serum bilirubin with plain agar: a meta-analysis. AB - Neonatal jaundice is one of the most common problems leading to prolonged hospitalization of the newborn. Therefore, an effective low-risk, low-cost therapy reducing hospitalization is highly desirable. Plain dried agar, an extract of seaweed, is low cost and low risk; it can bind bilirubin in the gut, decreasing its enterohepatic circulation, thereby decreasing serum levels. Because of conflicting conclusions in the studies of agar's effectiveness in the prevention and treatment of neonatal jaundice, a meta-analysis of their methodologies was done. Criteria for assessing prospective controlled therapeutic trials of agar were established in six areas: hypothesis and clinical outcome, patient selection, treatment group allocation, therapeutic maneuver, use of cotherapies, and data analysis. Nine prospective clinical trials of agar therapy were evaluated using these six criteria. The seven studies with negative conclusions regarding agar's efficacy failed to meet the criteria in several categories: patient selection, therapeutic maneuver, use of cotherapies, or data analysis. All of the studies, including the positive studies, were at risk for biased treatment allocation. Although the pooled data analysis suggests that prophylactic agar treatment is associated with reduced peak serum bilirubin levels, this observation must be interpreted cautiously in light of heterogenous patient populations and the methodologic problems described. Based on this meta analysis, agar therapy for neonatal jaundice can neither be recommended nor rejected. The methodologic analysis gives clear guidance for future research concerning the effectiveness of agar in treating neonatal jaundice and provides a model for meta-analysis of other prospective trials in pediatrics. PMID- 3050868 TI - Laser therapy for selected cutaneous vascular lesions in the pediatric population: a review. AB - Three cutaneous vascular lesions of childhood (the spider angioma [nevus araneus], the strawberry hemangioma, and the port wine stain) are reviewed, with particular emphasis on the present and future role of laser therapy in their management. PMID- 3050869 TI - [Validation of the use of protease inhibitors in severe forms of pneumonia in young infants]. PMID- 3050870 TI - Estimating the parameters of multidimensional signal detection theory from simultaneous ratings on separate stimulus components. PMID- 3050871 TI - Founders of pediatric pathology: Sidney Farber 1903-1973. PMID- 3050872 TI - The Sidney Farber memorial lecture--creative ideation and inquiry in developmental pathology. PMID- 3050873 TI - Persistent pulmonary hypertension of the newborn: pulmonary pathologic aspects. PMID- 3050874 TI - Some problems in the meanings of terms, as exemplified by tubulointerstitial and medullary cystic diseases of the kidneys. PMID- 3050875 TI - Anatomic basis for tomographic analysis of the pediatric heart at autopsy. PMID- 3050876 TI - Pathology of hearts with a univentricular atrioventricular connection. PMID- 3050878 TI - Giuseppe Moruzzi, 1910-1986. PMID- 3050879 TI - Baccalaureate and master's degree programs in nursing accredited by the NLN 1988 89. PMID- 3050877 TI - Effect of various types of acute exercise and exercise training on the insulin sensitivity of rat soleus muscle measured in vitro. AB - Effects of acute exercise varying in duration and intensity, as well as of two training regimes (endurance and sprint training) on the sensitivity of the soleus muscle of rat to insulin was measured in vitro and compared in rats. As an index of the muscle insulin sensitivity the hormone concentration in the incubation medium which would produce half maximum stimulation of lactate production (LA) and glycogen synthesis was determined. A single bout of moderate endurance exercise (60 min treadmill running at 20 m x min-1, 0 degrees inclination) increased the rate of LA production at the hormone concentrations used and increased the sensitivity of the process to insulin at 0.25 and 2 h but not 24 h after termination of exercise. Similar though less pronounced effects were found after heavy endurance exercise (30 min at 25 m x min-1, 10 degrees), but sprint exercise (6 x 10 s bouts at 43 m x min-1, 0 degrees) had no influence on the insulin sensitivity of the soleus muscle. The rate of glycogen synthesis in vitro was accelerated after endurance exercise, but the sensitivity of this process to insulin was unaffected by the preceding exercise. Endurance training for 5 weeks caused marked enhancement of sensitivity of both LA production and glycogen synthesis to insulin, which persisted for at least 48 h after the last training session. No changes in the soleus muscle sensitivity to insulin were found after sprint training. It is concluded that the increased insulin sensitivity of glucose utilization by skeletal muscle which occurs after endurance exercise and particularly during endurance training can substantially contribute to improved carbohydrate tolerance.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3050880 TI - Baccalaureate education in nursing: key to a professional career in nursing--BRN 1988-1989. PMID- 3050881 TI - Educational outcomes: assessment of quality--a compendium of measurement tools for baccalaureate nursing programs. PMID- 3050883 TI - Tools for measuring performance outcomes in baccalaureate nursing programs. PMID- 3050884 TI - Tools for measuring affective outcomes in baccalaureate nursing programs. PMID- 3050885 TI - Educational outcomes: assessment of quality--a compendium of measurement tools for associated degree nursing programs. PMID- 3050882 TI - Tools for measuring cognitive outcomes in baccalaureate nursing programs. PMID- 3050886 TI - Tools for measuring cognitive outcomes in associate degree nursing programs. PMID- 3050888 TI - Tools for measuring affective outcomes in associate degree nursing programs. PMID- 3050887 TI - Tools for measuring performance outcomes in associate degree nursing programs. PMID- 3050889 TI - Tools for measuring cognitive outcomes in diploma nursing programs. PMID- 3050890 TI - Tools for measuring performance outcomes in diploma nursing programs. PMID- 3050891 TI - Tools for measuring affective outcomes in diploma nursing programs. PMID- 3050892 TI - Practical nursing programs accredited by the NLN 1988. PMID- 3050894 TI - The 5' coding region of the Plasmodium falciparum circumsporozoite protein gene is a focus for deletion and insertion events. PMID- 3050895 TI - Modified Birnboim-Doly method for rapid detection of plasmid copy number. PMID- 3050893 TI - Secondary structure of the leader transcript from the Escherichia coli S10 ribosomal protein operon. AB - Genetic analysis of the autogenous control of the S10 ribosomal protein operon of Escherichia coli has suggested that the secondary or tertiary structure of the leader transcript is important for this regulation. We have therefore determined the secondary structure of the leader by enzyme digestion and chemical modification. Our results suggest that the 172 base leader exists in two forms, differing only immediately upstream of the Shine-Dalgarno sequence of the first gene. We discuss the possibility that the equilibrium between these alternate structures is important for the L4-mediated regulation of translation of the S10 operon. We have also determined the structure of several mutant transcripts. Correlation of these structures with the regulatory phenotypes suggest that a hairpin about 50 bases upstream of the first gene is essential for the control of translation of the operon. Finally, our results show that a two base substitution in an eight base loop destabilizes the attached stem. PMID- 3050896 TI - RNA polymerase induces DNA bending at yeast mitochondrial promoters. AB - Yeast mitochondrial RNA polymerase can bind specifically to promoter-containing DNA fragments in vitro as detected by DNAse I or methidiumpropyl-EDTA. Fe(II) protection assays and gel retardation experiments. Retardation of RNA polymerase DNA complexes was most pronounced when the promoter was located in the middle of a DNA fragment and was diminished when RNA polymerase was bound near one of the ends. This indicates that upon RNA polymerase-binding the DNA undergoes a conformational change which is most likely a bend. The degree of introduced bending correlated with the efficiency of transcription and promoter-binding in a series of promoter mutants, suggesting that bending is a functional event during promoter utilisation. PMID- 3050899 TI - Nucleotide sequence of the aceB gene encoding malate synthase A in Escherichia coli. PMID- 3050897 TI - Molecular characterization of GCD1, a yeast gene required for general control of amino acid biosynthesis and cell-cycle initiation. AB - The GCD1 gene product of Saccharomyces cerevisiae has been implicated in the coordination of the cell cycle with the general control of amino acid biosynthesis (M. Wolfner et al., J. Mol. Biol. 96:273-290, 1975). Strains containing the gcd1-1 allele constitutively express the amino acid biosynthetic genes at the induced levels normally found only during conditions of amino acid starvation. In addition, gcd1-1 strains do not grow at high temperatures because under these conditions they are unable to proceed beyond the START step of the cell division cycle. We have cloned and sequenced the GCD1 gene and examined various aspects of cellular metabolism in order to elucidate its role(s) in regulating gene expression and the cell cycle. GCD1 encodes a 1.7 kb RNA whose expression is not regulated as a function of amino acid starvation. Overexpression of this RNA does not affect the regulation of amino acid biosynthetic genes or cell growth. GCD1 is an essential gene because cells containing a gcd1-HIS3 disruption are unable to grow. The essential function of GCD1 may be involved in protein synthesis because a gcd1-1 strain incorporates low levels of 35S-methionine into protein when cells are shifted to the restrictive temperature. GCD1 encodes a protein of 511 amino acids whose predicted sequence does not exhibit significant homology to any other known proteins and appears too large to be a ribosomal protein. We suggest that GCD1 encodes a component of the normal protein synthesis machinery that is involved in the translational regulation of GCN4, a protein that coordinately activates the transcription of amino acid biosynthetic genes. GCD1 may also be part of a sensing mechanism in which cells monitor the protein synthesis capacity prior to initiating a new cell division cycle. PMID- 3050898 TI - Mutational study of the rRNA in yeast mitochondria: functional importance of T1696 in the large rRNA gene. AB - Four intragenic suppressors of a mitochondrial mutation in the 21S rRNA gene have been characterized in S. cerevisiae. The determination of the nature of the nucleotide changes in the suppressor strains showed that a T at position 1696 in the large rRNA gene is essential for correct function of the mitoribosome. The importance of this specific nucleotide and the fact that this mitochondrial mutation can also be suppressed by a mutation in a nuclear gene are in good agreement with a rRNA-r protein interaction in this part of domain IV, which functional importance is demonstrated in vivo by our results. PMID- 3050900 TI - For Queen and Country. PMID- 3050901 TI - [Leukemia in childhood]. AB - The prognosis of leukemia in children has changed remarkably in the last 20 years. Today more than 50% of children with Acute Lymphoblastic Leukemia (ALL) and about 30% of children with Acute non Lymphoblastic Leukemia (ANLL) can be cured with chemotherapy. The German Group BFM has obtained a significant improvement of results, both in ALL and ANLL using multidrug intensive treatment schedules. In Italy, thanks to the Italian Pediatrics Association of Hematology and Oncology (AIEOP), results have been improved in the last 10 years; very recently, new protocols with the BFM strategy have been started. Allogenic matched bone marrow transplantation (BMT) is indicated in children with ALL in 2nd complete remission (CR) following a relapse during or shortly after discontinuing treatment and in patients with Chronic Myeloid Leukemia. Chemotherapy results remain very poor in these patients. Allogenic BMT in usually performed also in children with ANLL in 1st CR. Autologous BMT, and allogenic BMT mismatched or from unrelated donors are being used with promising results when matched donors are not available. Most children cured of leukemia can enjoy a normal quality of life. However long term studies are still needed to determine the incidence of late effects, and to evaluate the psychosocial impact of the disease. In this context is becoming more and more important the role of the family doctor. PMID- 3050902 TI - [Pro-allergy role of infection. A component of the mode of reacting]. AB - No organism from the cradle lives germ-free or allergen-free. The modalities by which infections facilitate conditions of abnormal reactivity, in particular respiratory asthma and allergy to cow milk proteins, are examined. 216 asthmatic children and 50 infants with rotavirus enteritis have been considered. Infections, besides representing stimuli directly projected on the immune system, also constitute factors which more generally influence the way of reacting. PMID- 3050903 TI - Stereoselectivity: an issue of significant importance in clinical pharmacology. AB - Many drugs have one or more asymmetric centers and are administered as a racemate containing an equal mixture of enantiomers with different pharmacologic properties, routes, and rates of disposition in humans. For example, S-warfarin is more potent than R-warfarin, and is metabolized by different pathways. The S enantiomer is primarily oxidized, and the R-enantiomer is metabolized by both oxidation and reduction. Nevertheless, because of the difficulty in separating and analyzing individual enantiomers, most pharmacokinetic and pharmacodynamic studies on drugs have been performed without considering the stereochemical factors. This is unfortunate, because a nonstereoselective approach to the study of chiral drugs precludes insight into potential valuable information that may be relevant to drug development and evaluation. On the other hand, when the pharmacologic properties (including activity, disposition, and interaction with the other enantiomer) of enantiomers have been defined, manipulation of the enantiomeric ratio or use of the pure enantiomer can be pursued to optimize therapeutic efficacy. PMID- 3050904 TI - Duration of effect of gallopamil, a calcium channel blocker, on methacholine induced bronchoconstriction. AB - We previously demonstrated a modest but significant protective effect of inhaled gallopamil (D600), the methoxy derivative of verapamil, against methacholine induced bronchoconstriction; however, the duration of the protective effect of this and other calcium channel blockers is unknown. We therefore evaluated the duration of this protective effect in 15 asthmatic subjects in a prospective, placebo-controlled trial. Methacholine challenges (Cockcroft method) were performed 2 hours before and 30 minutes after the administration of placebo, and 1 mg and 10 mg of inhaled gallopamil. Gallopamil did not alter resting airway caliber, but significantly increased the concentration of methacholine required to decrease the FEV1 20% 30 minutes after the dose. Results with both the 10-mg and 1-mg doses were significantly different from placebo, but not from each other. The duration of this protective effect was transient in the group as a whole; the mean drug activity ratios were not significantly different with this sample 2.5 hours after the dose. Thus the short duration of effect limits the potential clinical usefulness of gallopamil in suppressing the signs and symptoms of chronic asthma. PMID- 3050905 TI - Preoperative flurbiprofen in oral surgery: a method of choice in controlling postoperative pain. AB - The analgesic efficacy of a single 50-mg preoperative dose of flurbiprofen was compared with ACC-30 (aspirin 375 mg, codeine 30 mg, caffeine 30 mg) and a placebo. Forty patients scheduled for the surgical removal of impacted maxillary third molars were enrolled in a double-blind, randomized study. Using a within subject design we compared the analgesic efficacy of (1) preoperative flurbiprofen 50 mg with placebo in 20 patients, and (2) preoperative ACC-30 with placebo in 20 other patients. Using a between-group design, we then compared the analgesic efficacy of (3) each drug given preoperatively and postoperatively, and (4) each drug given postoperatively only. Patients rated 2 pain dimensions, intensity and unpleasantness, hourly for 8 hours after the presurgical dose. The results of this study indicate that better analgesia was obtained when flurbiprofen was given preoperatively compared to only after surgery. Conversely, preoperative administration of ACC-30 did not demonstrate any significant influence on postsurgical analgesia. When comparing the 2 drugs, flurbiprofen proved to be superior in providing pain relief only when it was given prior to surgery. There was no difference between them when given only after surgery. Side effects were moderate and not significantly different between patients receiving flurbiprofen and those receiving ACC-30. PMID- 3050906 TI - The pain intensity at analgesic intake, and the efficacy of diflunisal in single doses and effervescent acetaminophen in single and repeated doses. AB - A double-blind, randomized analgesic trial was carried out in 150 patients undergoing surgical removal of their 2 impacted lower wisdom teeth. The analgesic efficacy of effervescent acetaminophen 500 or 1000 mg in a 2-dose regimen was compared with that of diflunisal 500 mg in a single dose. Each dose was taken when subjectively needed and the pain intensity was measured on a visual analog scale during the 10-hour period after first medication. The best pain reduction was achieved with diflunisal. The difference between diflunisal 500 mg and acetaminophen 1000 mg was significant, as was that between acetaminophen 1000 and 500 mg. The peak effect after the first dose occurred later but was greater with diflunisal than with acetaminophen. Patients needing analgesics at low pain intensities seemed to discriminate better between treatments, and the efficacy of acetaminophen was weakly dependent on the initial pain intensity. This intensity was difficult to predict, and only a poor correlation was found between the initial pain intensity and the patient's prior estimate of this. PMID- 3050907 TI - Phototoxic properties of quinine and quinidine: two quinoline methanol isomers. AB - Clinical photoreactions have been reported for quinine and quinidine after systemic and topical administration. We have investigated the phototoxic properties of these two quinoline methanol isomers in vitro using the Candida albicans inhibition test and photohemolysis, and in vivo with the mouse tail phototoxicity test. Both isomers were phototoxic in the hemolysis model, quinine being the more potent compound. In the Candida test only quinidine was phototoxically active. In the mouse tail model, measuring edema, the phototoxic activity of quinidine was comparatively low, causing a 7.3% wet weight increase of tail tissue at a dose of 150 mg/kg intraperitoneally of drug and 54 J/cm2 of UVA. In spite of its structural similarity to quinidine, quinine was not phototoxic in the mouse. These studies support the assumption, based on clinical data, that quinine photoreactions probably have a non-phototoxic mechanism. For quinidine, however, light-induced reactions based on phototoxicity can not be ruled out, since low-grade phototoxic properties were demonstrated in vivo. PMID- 3050908 TI - Amiodarone-induced pseudoporphyria. PMID- 3050909 TI - Immunocytochemical demonstration of insulin or insulin-like immunoreactivity in the rat prostate gland. AB - The indirect immunocytochemical technique was used in conjunction with anti insulin antisera to localize insulin-like immunoreactivity in tissue sections and primary cell cultures from rat prostate gland. Positive immunostaining for insulin-like reactivity was demonstrated in the epithelium of the prostate gland. There appeared to be some variation in the intensity and localization of the immunoreactivity within different regions of the gland. Prostatic epithelial cells grown in culture in an insulin-free medium displayed strong cytoplasmic immunostaining when treated with anti-insulin antisera, while nuclear staining was absent. These results demonstrate that insulin-like immunoreactivity is present in the epithelium of the prostate gland and suggest that there may be some local insulin or insulin-like synthesis in this organ. PMID- 3050910 TI - Significance of duct-acinar dysplasia in prostatic carcinogenesis. AB - Duct-acinar dysplasia of the prostate is defined as a premalignant lesion characterized by cytologic, and especially nuclear, abnormalities that resemble those of carcinoma and affect the lining epithelial cells of preexisting ducts and acini. A progressive continuum of cytologic abnormality is divided into three grades, with grade III identified by the presence of numerous large nucleoli. Foci of carcinoma have been identified at their point of origin from dysplastic ducts, which confirms the malignant potential of this lesion. Foci of dysplasia are found in roughly 80% of prostates with invasive carcinoma and 40% of glands without cancer. It is concluded that the majority of prostate cancers probably arise within dysplasia foci. Foci of grade III dysplasia occasionally evolve by proliferation of cells to fill duct lumens. The intraluminal cell masses display a cribriform pattern, which has previously been interpreted as a variant of invasive Gleason grade 3 carcinoma. Dysplasia is further linked biologically to malignant transformation by the immunohistochemical demonstration of progressive loss of cytoplasmic differentiation markers and inappropriate overexpression of some markers during the evolution of dysplasia. PMID- 3050911 TI - Symptomatic relief following carpal tunnel decompression with normal electroneuromyographic studies. PMID- 3050912 TI - Intraarticular trochanteric wire migration following bilateral total hip arthroplasty. A case report. AB - A 64-year-old man underwent bilateral simultaneous total hip replacement and experienced articular interposition of a wire fragment in the right hip joint 3 years postoperatively. Because of destructive wear of the acetabulum, this eventually required revision total hip arthroplasty 12 years after the original surgery. PMID- 3050913 TI - Perinatal lethal osteogenesis imperfecta: radiologic and pathologic evaluation of seven prenatally diagnosed cases. AB - The radiologic and pathologic characteristics of 7 cases of lethal osteogenesis imperfecta (OI), diagnosed prenatally by ultrasound in the 15th to 34th week, are described. They include four variants of the Sillence classification: types IIA, IIB, IIC, and type III. The radiologic criteria that differentiate these types of OI are described. The histopathology of the bones differed only slightly in types IIA, IIB, and III; OI type IIC, however, differed markedly from the other types. PMID- 3050914 TI - Nephrogenic metaplasia of urinary tract in children: report of three cases and review of the literature. AB - Nephrogenic metaplasia of the urinary tract, originally thought to be a benign tumor with possible malignant potential, is commonly called nephrogenic adenoma. It predominantly affects male adults and is rarely seen in children. In this report 18 pediatric cases are reviewed and some clinical and pathologic parameters are compared with the condition in adults. Male to female ratio is reversed (3.7:1 in adults and 1:3.5 in children). Recurrences are more common and are more frequently multiple in the pediatric age group. Although no malignant transformation has been reported, prolonged follow-up is recommended since the natural history of this lesion is still uncertain. PMID- 3050915 TI - The general pediatrician as care coordinator for children with chronic illness. AB - As the number of children with chronic illness increases due to advances in medical technology, general pediatricians are faced with the challenge of providing continuing care for such patients. These children and their families are most in need of a care coordinator to guide them through the complexities of obtaining optimal care in all aspects: medical, emotional, social, and developmental. The primary pediatrician is logically positioned to assumed the role of care coordinator. This undertaking requires a knowledge of the needs of such children and their families, an ability to interact with other professionals as member of a team, a sensitivity to the overall functional status of the child and family, and a commitment of a large amount of time and effort. Although these skills are rarely taught during residency training, they can be acquired through continuing education, thereby allowing the pediatrician to experience the satisfaction that derives from helping someone truly in need. PMID- 3050916 TI - Chronic illness and early development. The parent's perspective. AB - While many chronic illnesses are diagnosed in infancy, there is little systematic study of the impact of chronic illness on early social development and how parental adjustment to a chronically ill infant contributes to subsequent outcomes. This paper outlines some of the factors which affect parents' ability to care for chronically ill infants and preschoolers. It draws upon studies of infants with other medical problems, ongoing longitudinal study of infants with cystic fibrosis and congenital heart disease by the authors; and the authors' clinical experience. PMID- 3050917 TI - Meningomyelocele. Assessment and management. AB - Failure of the neural tube to close in a fetus is one of the most devastating and yet common congenital malformations. Various organ systems including the skeletal, muscular, urologic, digestive, respiratory, skin and central nervous systems are involved. This involvement results in complicated and interrelated problems requiring services from the medical specialties of pediatrics, orthopedics, urology and neurosurgery; the allied health professions of physical therapy, occupational therapy, speech pathology, nutrition, social work and nursing; and the psychoeducational professions of education and psychology. This article attempts to summarize the information about the current needs and management techniques of children with meningomyelocele for the general pediatrician who is frequently called upon to play an important role in coordination and communication with the families and other professionals caring for these children. PMID- 3050918 TI - Psychosocial management of chronic lung disorders. AB - Chronic lung disorders are increasingly prevalent and account for the most of the disabilities and deaths in infants and young children. The pediatrician has a unique opportunity in the early detection, counseling and referral of psychosocial problems related to chronic lung disease, as well as in assuring continuity of appropriate medical care. The present paper outlines ways the pediatrician might effectively address psychological or behavioral aspects of management of several common pediatric pulmonary disorders, namely, asthma, cystic fibrosis, bronchopulmonary dysplasia and infant tracheostomy. PMID- 3050919 TI - Advances in ulcerative colitis. AB - Ulcerative colitis is one of the two common chronic inflammatory bowel diseases which affect the colon of children. The disease can occur at any time during infancy and childhood and is far commoner than Crohn's disease of the colon in children less than 6 years old. The Jewish population outside of Israel is at far greater risk of developing the condition than any other ethnic group. The reason for this is unknown. The chances of a family member developing the condition is 2 3 times as great as in the general population. The etiology of the condition remains unknown; however, recent advances in the understanding of the immune mechanisms in the bowel and circulation indicate there are major immunological differences between ulcerative colitis and Crohn's disease. Intestinal B cells secrete enormously increased amounts of IgG1 and a lesser increase in IgG3 in ulcerative colitis whereas in Crohn's disease, all IgG subclasses are increased, but especially IgG2. Failure of the gut immune system to control antigen crossing the colonic mucosa may be the basis for the condition. The disease is classified as moderate to severe two thirds of children as opposed to less than one third of adults. Diagnostic testing must include 3 stool cultures negative for bacterial and viral pathogens, 3 stools negative for amebiasis, trichuriasis and other intestinal parasites and absence of clostridium difficile and its toxin in the stool. Flexible proctosigmoidoscopy and/or colonoscopy should be done in every case with biopsies. Barium enema is contraindicated in the severely ill patient. Major improvements in medical treatment being tested involve the development of nonabsorbable corticosteroid enemas and sulfapyridene-free forms of salicylazosulfapyridene for use in enema and oral form. Surgery for ulcerative colitis has made major advances with the development of the Koch pouch (internal ileostomy) and ileoproctostomy. Both procedures although associated with relatively high complication rates, are esthetically and psychologically better than standard ileostomy because in neither procedure must the patient wear an ileostomy appliance. However these advanced surgical procedures are typically not done until adolescence is reached. PMID- 3050921 TI - Cerebral palsy: a 1987 perspective. AB - Changing views on reaching a diagnosis, methods of assessment, and management are apparent for neurological disorders in general, and cerebral palsy in particular. A personal approach to the problems associated with the evaluation and care of children and young people with cerebral palsy is outlined, with particular emphasis on the contributions form, and optimal use of, the multidisciplinary assessment team. PMID- 3050922 TI - Rheumatic disease in childhood. AB - The autoimmune diseases, juvenile rheumatoid arthritis and systemic lupus erythematosus, are two of the more common acquired chronic diseases of childhood. In this article we will describe features of the diagnosis and treatment of these disorders. Particular emphasis is placed on the complications and how to anticipate them. The management of these diseases must include an appreciation of how chronic illness can affect family function and school relationships. With early attention to these nonmedicinal details, the majority of children can progress through childhood and adolescence with few physical or emotional handicaps. PMID- 3050920 TI - Essential concepts in the care of children with chronic illness. AB - Children with chronic illness constitute an increasingly significant segment of the child population seeking care in Western industrialized countries. There are concerns that traditional disease-specific ways of thinking about these children and organizing their care may not meet the real life needs of the children and their families. This article examines a series of concepts that are fundamental for the humane and biomedically sound care of children with chronic illness, and a model of care is proposed. Relevant data from one program are presented. PMID- 3050923 TI - Diabetes mellitus. AB - The diagnosis of diabetes mellitus implies a life-time regimented by insulin therapy, dietary manipulations and monitoring glycaemic control. There is also the ever present risk of ketosis and hypoglycaemia and as the child grows up there is an increasing awareness of the later complications of diabetes. This paper is concerned with the care of the child with diabetes from diagnosis, within the context of his home and school, with an emphasis on the emotional response of the child and his family. PMID- 3050924 TI - How families cope with diabetes in adolescence. An approach and case analyses. AB - In this paper we describe our newly constructed Family Coping Coding System. This scheme was constructed to identify family coping strategies that involve appraisal, problem solving, and emotion management dimensions. We discuss the theoretical rationale, meanings and reliability of the coping codes, and illustrate them through excerpts drawn from family discussions of a recent stressful situation (the onset of a chronic or acute illness in an adolescent member). Finally, we consider the clinical research relevance of this new assessment technique, exemplifying this potential with respect to medical compliance. We present analyses of two families with diabetic adolescents who strikingly differ with respect to compliance, and explore which family coping strategies may be predictive of an adolescent's favorable or problematic compliance to diabetes management. PMID- 3050925 TI - Psychosocial factors in childhood diabetes and seizure disorders. The family approach. AB - Not infrequently chronic childhood illnesses such as diabetes and seizures disorders are not as well controlled as might be expected by standard medication, due to a wide range of adverse psychosocial factors. These may be a reaction to chronic ill health or coincidental. In either case it is vital to understand the interactions between the child, his family, the illness and environment if the failure to control the disorder is to be overcome. This paper outlines the family approach to assessment and management of chronic ill health in children. PMID- 3050926 TI - Rheumatic disorders in adults under 50. Guidelines for differential diagnosis. AB - Rheumatic disorders are not uncommon in patients between 20 and 50 years of age, and the differential diagnosis may be difficult. However, after a careful history and thorough physical examination, the cause usually becomes apparent. Laboratory findings alone should not be relied on for diagnosis. Because the impact on younger adults may be devastating and the potential disability may be present for many years, these patients represent an important challenge for any practicing physician. PMID- 3050927 TI - Systemic infections affecting the liver. Some cause jaundice, some do not. AB - The patient who has clinical jaundice, abnormal results on liver function tests, or both presents a difficult diagnostic challenge. Many infectious diseases affect the liver, and the extent of involvement determines the degree of clinically apparent jaundice. Some diseases that affect the liver minimally cause no jaundice at all. An important clue to the cause of the disorder is the pattern of abnormal results on liver function tests. Increased alkaline phosphatase predominates with Q fever, secondary or tertiary syphilis, clonorchiasis, and hepatic candidiasis, while elevated levels of serum transaminases characterize viral hepatitis, leptospirosis, mononucleosis syndromes, legionnaires' disease, typhoid fever, toxic shock syndrome, and yellow fever. Increases in serum bilirubin are typical with jaundice caused by clostridial myelonecrosis, severe bacterial sepsis, and relapsing fever (borreliosis). These findings together with the patient's history, physical findings, and basic laboratory tests provide a presumptive diagnosis in most cases. PMID- 3050928 TI - Viral hepatitis. The alphabet game. AB - Differential diagnosis of viral hepatitis begins with a check for darkened urine and bile in the urine. These hallmarks of conjugated hyperbilirubinemia immediately rule out prehepatic liver disease. Next, studies are done for the elevated transaminase levels that are characteristic of hepatitis infection, and a thorough history is taken to rule out drug- and toxin-induced hepatitis that may mimic acute viral hepatitis. Elevated alkaline phosphatase is a good marker of cholestasis. Ultrasonography can clarify this diagnosis. The classic presenting symptoms of viral hepatitis are jaundice, nausea, vomiting, malaise, anorexia, and dull right upper quadrant pain. However, serologic studies are needed to detect the presence of specific viral agents. PMID- 3050929 TI - Noninfectious jaundice. Figuring out what's going on. PMID- 3050930 TI - Drug-induced jaundice. An uncommon but puzzling reaction. AB - Drug-induced jaundice is relatively uncommon but can be a diagnostic puzzle. Because so many pharmaceutical classes and individual agents can produce jaundice, a thorough history of medications taken should be obtained from a patient presenting with jaundice. Jaundice usually resolves when the offending agent is discontinued. PMID- 3050931 TI - Radiologic evaluation of the jaundiced patient. Diagnostic and therapeutic role of current procedures. AB - Radiologic evaluation of the jaundiced patient generally begins with ultrasound. Computed tomography may provide better information regarding the exact level of obstruction, but it is more expensive and time-consuming than ultrasound and carries the risk associated with intravenous contrast. It thus should be used only when ultrasound findings are likely to be inadequate. Percutaneous transhepatic cholangiography and endoscopic retrograde cholangiopancreatography are both relatively safe, reliable procedures for direct opacification of the biliary tree. The choice depends on clinical and ultrasound findings. Percutaneous transhepatic cholangiography provides a foundation for percutaneous drainage if necessary. Cholescintigraphy in the jaundiced patient provides physiologic information but poor anatomic detail. It is useful in establishing common duct functional patency. PMID- 3050932 TI - Diabetes in the elderly. A unique set of management challenges. AB - The overall goals in regard to management of diabetes in the elderly are twofold. First, blood glucose control should be kept as near normal as possible (certainly always less than 200 mg/dl) without inordinately increasing risks related to therapy; this will decrease the risk of acute complications and hopefully of long term ones as well. Second, overall functional status should be maintained or improved. Although a number of factors peculiar to old age complicate the management of diabetes, with careful planning these goals should be attainable. PMID- 3050933 TI - Vertigo. How serious are recurrent and single attacks? AB - Through history taking and physical examination, the primary care physician should be able to distinguish between the four most common causes of recurrent vertigo. Particular caution is advised when dealing with the first attack of vertigo, because more sinister possibilities may be present, although most of the disorders that cause vertigo are benign. Categories of treatment that can be used include medical, rehabilitative or exercise-related, surgical, dietary, and unconventional (eg, acupuncture, manipulation). PMID- 3050934 TI - Do you favor ... routine neonatal circumcision? Yes. PMID- 3050935 TI - Do you favor...routine neonatal circumcision? No. PMID- 3050936 TI - Prolongation of the QT interval by ketanserin. AB - In hypertensive patients single doses of ketanserin 40 mg prolonged the corrected QT interval (QTc) for at least 8 hours, with a maximal increase of 35 ms (P less than 0.001, n = 6) after 2 hours. During chronic dosing (20 and 40 mg b.d.) the QTc was further prolonged, by 46 and 45 ms respectively. QTc prolongation after treatment with a mean dose of 73 mg/day for 7 weeks (n = 26) was significantly related to body weight (r = -0.58, P less than 0.01), and to the dose of ketanserin corrected for body weight (r = 0.63, P less than 0.01), but not to plasma concentrations of ketanserin, ketanserinol, potassium or calcium. High doses of ketanserin (mean dose 167 mg/day, n = 9) increased the QTc by 40 ms (P less than 0.001), with prolongation of up to 80 ms in individual patients. Treatment with ketanserin at doses proposed for clinical use (40-80 mg/day) may carry a risk of ventricular arrhythmias. PMID- 3050937 TI - The diagnosis and classification of scleroderma (systemic sclerosis). AB - Difficulty in the diagnosis of the disease scleroderma may occur at the early stage prior to the development of obvious skin sclerosis. A presumptive diagnosis may be made if Raynaud's phenomenon is accompanied by a positive 'neck test', 'scleroderma' capillary changes in the nailfolds or antinuclear antibodies. Definitive diagnosis may have to be delayed for several years from the onset of Raynaud's phenomenon until definite characteristic skin changes are seen. Ten cases in which an earlier diagnosis of scleroderma was not substantiated are listed. The earlier incorrect diagnosis would have been avoided by use of the methods described in this paper. Various terms have been used to denote subdivisions of scleroderma. These include acrosclerosis, diffuse scleroderma and CREST. We have used the terms Type 1, Type 2 and Type 3 based on the early extent of the skin sclerosis where Type 1 (limited extent) indicates sclerodactyly only, Type 2 (moderate extent) indicates sclerosis proximal to the metacarpophalangeal joints but excluding the trunk and Type 3 (extensive) indicates diffuse skin sclerosis including the trunk. The clinical value of this simple classification is reviewed and contrasted to other classifications which appear to be poorly defined and of limited use. PMID- 3050938 TI - Haemolytic anaemia associated with splenic haemangiomata. AB - We describe a 33 year old woman who developed unexplained haemolytic anaemia following renal transplantation and who was found to have multiple splenic haemangiomata. This case demonstrates that splenic haemangiomata may be a cause of haemolytic anaemia in the absence of abnormalities in coagulation. PMID- 3050939 TI - Recent advances in the understanding of dementia. PMID- 3050940 TI - Neurological complications of human immunodeficiency virus infection. AB - The protean neurological manifestations of human immunodeficiency virus (HIV) infection are reviewed. Both the central nervous system and peripheral nervous system may be affected and many of the complications may occur in individuals with acquired immunodeficiency syndrome (AIDS)-related complex, or who are seropositive for HIV alone as well as those with the established AIDS syndrome. Specific therapy is available for certain of these neurological conditions, but the clinical course in others is untreatable and progressive. Although it seems likely that the pathogenesis of some of these syndromes such as the AIDS-dementia complex are due to the direct effect of HIV on the nervous system, in others the neurological injury probably occurs as a consequence of the immunosuppression which HIV induces, or immune-mediated mechanisms. PMID- 3050941 TI - Effects of captopril in patients with chronic obstructive pulmonary disease and secondary pulmonary hypertension. AB - Ten patients with chronic obstructive pulmonary disease (COPD) with normal left ventricular function were given a single dose of captopril (0.25 mg/kg). Mean systemic arterial pressure, heart rate and coronary flow to the right ventricle were reduced significantly after captopril. In contrast captopril did not cause a significant fall in pulmonary arterial pressure or pulmonary vascular resistance. We conclude that captopril fails to produce haemodynamic benefits in such hypoxic patients and that angiotensin II is not playing a significant role in maintaining pulmonary vasoconstriction. PMID- 3050942 TI - Unstable angina. PMID- 3050943 TI - Graves' disease presenting as pyrexia of unknown origin. AB - Fever is a common clinical manifestation of inflammatory processes of the thyroid and thyroid crisis. On the other hand, fever alone as a presenting symptom of thyrotoxicosis, without other manifestations, is extremely rare. A female patient is described in whom fever persisted for two months prior to hospitalization, but without clinical symptoms or signs to lead to suspicion of thyroid disease. After exhaustive investigation it was found that the patient was suffering from hyperthyroidism. Fever disappeared gradually on antithyroid therapy, recurred when the drugs were withdrawn for a rechallenge trial, and cleared up again after renewal. Four other cases of persistent fever as a presenting symptom of hyperthyroidism were found on a review of previous publications. Thyrotoxicosis should, therefore, be included in the differential diagnosis of pyrexia of unknown origin. PMID- 3050945 TI - Characteristics of Medicare-eligible home care clients. PMID- 3050944 TI - Hypersensitivity reaction with intravenous GnRH after pulsatile subcutaneous GnRH treatment in male hypogonadotrophic hypogonadism. AB - Chronic pulsatile subcutaneous administration of low doses of gonadotrophin releasing hormone (GnRH) is an effective therapy for men with hypogonadotrophic hypogonadism. Hypersensitivity reactions to GnRH are rare. We wish to report hypersensitivity reactions with intravenous GnRH after low dose subcutaneous pulsatile GnRH treatment in two men with hypogonadotrophic hypogonadism due to suprasellar disease. PMID- 3050946 TI - Business methods in visiting nurse associations. 1926. By Dorothy Deming. PMID- 3050947 TI - The importance of higher education in nursing. PMID- 3050948 TI - [The CO diffusion capacityof the lung in the single breath version (DLCOSB)]. PMID- 3050949 TI - [Lung function and protease levels of BAL fluid in sarcoidosis II]. PMID- 3050950 TI - [Demonstration of inhalation injury]. PMID- 3050951 TI - [Frequent bronchial hyperreactivity as an expression of the burden of inhaled air pollutants in an urban region]. PMID- 3050952 TI - [[Are allergic bronchial diseases more common?]. PMID- 3050954 TI - [Detection of acarid allergens in the domestic environment]. PMID- 3050953 TI - [Diagnostic approach in occupational asthma]. PMID- 3050955 TI - [Increased lung volume in inhalant provocation]. PMID- 3050956 TI - [The spectrum of respiratory therapy in obstructive lung diseases]. PMID- 3050957 TI - [Indications for proton spin tomography and computerized tomography in chest diagnoses]. PMID- 3050958 TI - [Bronchial carcinoma with mediastinal lymph node metastases--an indication for surgery?]. PMID- 3050959 TI - [Objective measurement of compliance during treatment of sleep apnea with continuous positive pressure]. PMID- 3050960 TI - [The treatment of sleep apnea syndromes (SAS)]. PMID- 3050961 TI - [The current status of research of sex offenses in adolescence]. PMID- 3050962 TI - [Deafness from the viewpoint of the child and adolescent psychiatrist. 2: Social and emotional development, child psychiatric morbidity, significance of the family and social environment]. PMID- 3050963 TI - [Family therapy in residential treatment: conditions, chances and necessities]. PMID- 3050964 TI - [Hemangiopericytoma. Morphologic and immunohistochemical findings in 9 patients and review of the literature]. PMID- 3050965 TI - [Immunohistologic studies of 2 sarcomatoid differentiated renal cell cancers. A contribution to differential diagnosis]. PMID- 3050967 TI - [Agenesis of the trachea]. PMID- 3050966 TI - [Ochronosis oculi]. PMID- 3050968 TI - [Bernard Rudolph Konrad von Langenbeck (1810-1887)]. PMID- 3050969 TI - Transport of radioactive deoxyglucose by the isolated perfused human placental cotyledon as measured by the paired-tracer dilution method. Inconsistency of the transfer rates across the two trophoblast borders with the simultaneously measured rate of transplacental transfer. AB - The uptake of [3H]2-deoxy-D-glucose ([3H]2DG) by the maternal and fetal borders of the trophoblast was measured in the dually perfused human placental cotyledon by use of the paired-tracer dilution method. The uptake was estimated from the maximum extraction of the tracer. The transcellular component of transplacental transport of [3H]2DG was measured simultaneously. From the rates of uptake by the two trophoblast borders the rate of transcellular transport was predicted. The rate predicted was significantly lower than the rate observed. This was taken to indicate that the paired-tracer dilution method based on the maximum extraction underestimated the rates of uptake. PMID- 3050970 TI - Immunohistochemical detection of human placental cytochrome P-450-associated mono oxygenase system inducible by maternal cigarette smoking. AB - Several xenobiotic-metabolizing mono-oxygenase activities, in conjunction with the immunohistochemical localization of the respective cytochrome P-450 forms, were investigated in placentae from smoking and non-smoking women. The antibodies used in the immunohistochemical analyses were monoclonal antibodies (MAbs) 1-7-1 and 2-66-3, prepared against the 3-methylcholanthrene-induced and phenobarbital induced rat liver P-450, respectively. The mono-oxygenase activities were higher in placental microsomes from smokers, although no difference was observed in microsomal P-450 content. A distinct positive staining with MAb 1-7-1 was detectable in the trophoblastic layer of all placentae from smokers. In placentae from non-smokers, minimal cytoplasmic staining was observed in large villi in a few trophoblastic cells. The staining reaction was negative when MAb 2-66-3 or non-specific antibody were used. These results demonstrate that maternal cigarette smoking induces in the trophoblastic layer of the placenta a cytochrome P-450 form which is detectable immunohistochemically with the monoclonal antibody to 3-methylcholanthrene-inducible P-450 in rat liver. PMID- 3050971 TI - Fetal and maternal blood volumes in shed human placentae: discrepant results comparing morphometry to haemoglobin content. AB - The maternal blood volume (MBV) and fetal blood volume (FBV) of shed human placentae delivered by caesarean section at term were measured using a morphometric technique and from the placental content of adult and fetal haemoglobin. MBV was 35.3 +/- 1.5 (s.e.m.) per cent by the former and 11.0 +/- 1.5 per cent by the latter technique. FBV was 11.0 +/- 0.7 and 9.0 +/- 0.6 per cent respectively (n = 6). Measurement of the dimensions of individual villi initially photomicrographed in 0.9 per cent NaCl and subsequently re-photographed in fixative suggested that individual villi shrank to 0.7 of their initial volume during fixation. It is suggested that morphometric measurement of MBV may lead to approximately a threefold overestimate because of relative MBV expansion and villous tissue shrinkage during the process of fixation without alteration in overall placental volume. PMID- 3050973 TI - Eradicating fetomaternal fluid shift during perfusion fixation of the human placenta. AB - Perfusion fixation can redilate the fetal capillary network of the freshly delivered human placenta to its in vivo dimensions. As part of the fixation procedure in earlier experiments a venous back-pressure of 20 mmHg was applied on the rationale that this was necessary for full distension of the fetal capillaries. In order to verify this assertion, five mature placentae were fixed by perfusion fixation but without the application of such a back-pressure. No statistically significant difference was found between the values for the various parameters measured in placentae fixed in this manner compared with those processed using the original technique. In view of this finding, and of the implication that impedence to venous flow may cause fetomaternal fluid shift during in vitro perfusion experiments, it would appear that venous back-pressure should best be omitted from the perfusion fixation technique. The shrinkage rate associated with this technique was found to be approximately 2.0 per cent, based on the measurement of maternal erythrocyte diameters. PMID- 3050972 TI - Mouse and rat placental cell lines express abundant amounts of laminin. AB - Placental cell lines derived from midgestation placentae of outbred mice and rats were evaluated for the expression of the extracellular matrix protein laminin. The murine cell line, which has not been previously reported, demonstrates morphological characteristics similar to those of the rat cell line. Placental cell lines grow vigorously both in vitro and when transplanted to the peritoneum of allogeneic hosts. When transplanted, placental cells form cyst-like structures (with acellular cores) suspended in the peritoneal fluid, and invade abdominal structures forming solid masses. Using immunohistology, laminin was identified within in vitro cultured cells, within cyst-like structures and their acellular cores, and as a major component of the extracellular matrix of solid masses. Laminin was also identified in the normal rat chorioallantoic placenta. Evaluation of extracts from in vitro cultured placental cells, transplanted placental cells, and the normal chorioallantoic placenta by electrophoresis and immunoblotting demonstrated that laminins were composed of two species with molecular weights of 400,000 (A-chain) and 200,000 (B-chains). Mouse and rat placental cell lines may be valuable for studying laminin biosynthesis and function in the developing placenta. PMID- 3050974 TI - Placental handling of trace elements. PMID- 3050975 TI - [Isolation of spheroplasts from Escherichia coli 85 for aspartate-ammonia-lyase localization]. AB - Conditions were determined for preparation of spheroplasts from E. coli under the action of lysozyme in the presence of EDTA. The preparation took from 10 to 15 min. The degree of conversion to spheroplasts was monitored spectrophotometrically at 660 nm. The spheroplasts formed were unstable in Tris HCl buffer and immediately lysed, but they were more stable in 1 M sucrose. At lysozyme concentrations above 40 micrograms/ml of the reaction mixture, the cells lysed to a greater extent. The distribution of aspartate ammonia-lyase activity between the precipitate of the spheroplasts and the supernatant allowed the authors to suggest that aspartase should be located in the cytoplasm. PMID- 3050976 TI - Glucose stimulation of pancreatic islet blood flow by redistribution of the blood flow within the whole pancreatic gland. AB - Whole pancreatic blood flow (PBF) and islet blood flow (IBF) were measured 5 min after administration of glucose at serum glucose concentrations ranging between 8.5 and 25 mM. The serum insulin concentration increased proportionally (p less than 0.01) to the serum glucose concentration. PBF was unaffected by i.v. glucose administration at 5 min, while IBF increased in proportion to the serum glucose concentration of the animals in the range of 8.6-16.7 mM. No further increases in IBF could be seen at higher glucose concentrations. It is concluded that glucose preferentially stimulates IBF and that this increase, to a major extent, is due to a redistribution of flow within the gland. PMID- 3050977 TI - Clonal growth of immunocytochemically identified islet beta and alpha cells in culture. AB - The proliferative capacity of individual immunocytochemically identified islet beta cells was investigated in tissue culture. The pancreatic digest was filtered through a 20-micron filter to eliminate partially digested tissue fragments and islets; it was then cultured at low density to allow assessment of single cells. The type of cell was identified immunocytochemically by reaction with antibody to insulin or glucagon, and DNA synthesis was estimated from autoradiographs after incorporation of tritiated thymidine. Single immunocytochemically reactive beta or alpha cells attached to the culture substratum and then proliferated, directly proportional to time in culture. Single beta cells did not incorporate thymidine after 1 day in culture; nonendocrine cells, presumed to be mainly fibroblasts, readily incorporated thymidine. Beyond the first day, about 10% of the beta cells incorporated thymidine. The number of radioactively labeled, immunocytochemically reactive beta cells increased for 5 days in culture and then remained at the elevated level until experiments were terminated at 12 days. DNA synthesis in beta cells occurred in two waves separated by about 3 days, suggesting a generation time of about 3 days for immunocytochemically identified beta cells. Fetal calf serum was found to be an essential culture medium ingredient to sustain thymidine incorporation; horse serum was found to be an unsuitable substitute. Mitotic figures were recorded in differentiated beta and alpha cells. These studies conclusively show that differentiated beta and alpha cells can proliferate in culture.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3050978 TI - Effects of hydrocortisone on glucose- and cholecystokinin-induced insulin release from the isolated perfused rat pancreas. AB - The acute and chronic effects of hydrocortisone on insulin secretion were examined in the isolated perfused rat pancreas. In the first part of this study, the chronic effects of hydrocortisone on insulin release were examined using isolated perfused pancreas prepared from rats that had been given subcutaneous injections of hydrocortisone at doses of 1.25, 2.5, 5.0, and 10.0 mg/kg body weight once daily for 7 days. Hydrocortisone treatment led to a dose-dependent increase in insulin secretion in response to 8.3 mM glucose. The insulin response to 100 pM cholecystokinin (CCK-8) was also significantly higher in the hydrocortisone-treated rats than in the control group. However, the increment of insulin level over the value before CCK-8 addition in rats treated with hydrocortisone was not significantly different from that in the control rats. In the second part, the acute effects of hydrocortisone on insulin release were studied. Hydrocortisone (17-hydroxycorticosterone) at a concentration of 100 microM caused significant inhibition of the stimulatory effect of CCK-8 on insulin secretion. The inhibition started within 1 min of the beginning of hydrocortisone administration and ceased immediately after the termination of its infusion. We have demonstrated in this study a dual effect of hydrocortisone on insulin release: first, the potentiation of the insulin secretion stimulated by glucose but not by CCK-8 and, second, the inhibition of CCK-8-stimulated insulin secretion. PMID- 3050979 TI - Pancreatic trophism following colectomy in rats: the potential role of gastrointestinal hormones. AB - It has been shown that the large bowel contains substances with a potential to inhibit exocrine pancreatic function. Following large bowel removal in rats, there is an increase of pancreatic weight, digestive enzyme concentration, and secretion capacity in vitro. To evaluate the role of various GI hormones in the exocrine pancreatic adaptation following colectomy, we measured plasma cholecystokinin (CCK), neurotensin, glucagon, and insulin after meal stimulation. The test meal was applied via a transabdominal gastric tube in eight colectomized Wistar rats after a median of 18 days following surgery. Ten rats with a gastric tube without previous bowel surgery served as controls. After large bowel removal, there was impaired glucose tolerance and attenuated plasma insulin secretion. Baseline plasma glucagon levels were increased after colon removal, whereas the total postprandial glucagon release was decreased. Baseline and postprandial neurotensin values were comparable in both the experimental and control animals. Baseline and postprandial CCK plasma levels were intensely increased in the colectomized rats. It is assumed that the baseline and postprandial CCK pattern in rats after subtotal colectomy is responsible for exocrine pancreatic adaptation. PMID- 3050980 TI - Characterization and control of pulsatile secretion of insulin and glucagon. AB - Periodic oscillation of insulin and glucagon by isolated mice islets has been studied. Pulsatile secretion of insulin and glucagon was observed at all glucose concentrations tested. The frequency of oscillation per 20 min for glucagon was 5.0 +/- 0.26 and for insulin 4.0 +/- 0.26 (n = 6), approximating to periodicities of 4 and 5 min, respectively. These did not change by increasing the glucose concentration to 11.1 or 22.2 mM from 5.5 mM (basal). The maximal amplitude of glucagon secretion was not altered by raising the glucose concentration to 11.1 mM from basal. However, 22.2 mM glucose significantly suppressed the amount of glucagon released when compared with glucagon secretion in the presence of 5.5 mM glucose. In contrast, the maximal amplitude of insulin increased from 444.2 +/- 37.7 to 777.2 +/- 61.4 and from 271.8 +/- 35 to 701 +/- 26.5 pg/min (p less than 0.01, n = 6) by switching from basal to 11.1 and 22.2 mM glucose, respectively. We conclude from this study that the pacemaker controlling pulsatile secretion of insulin and glucagon is within the islet. Although the amplitude of secretion of these hormones is regulated by the ambient glucose concentration, the frequency of their pulsatile secretion is not. PMID- 3050982 TI - The familial aggregation of Alzheimer's disease: an epidemiological review. AB - This report reviews current data on the familial aggregation of Alzheimer's disease (AD). Single pedigree reports indicate that in few families AD is inherited as an autosomal dominant single gene disorder. Family studies show that first-degree relatives of AD patients have a higher lifetime incidence of AD than the general population or groups of nondemented subjects. Case-control studies indicate that the risk of developing AD is significantly higher for subjects with family members affected by dementia than for those without. The concordance rate in monozygotic twin pairs was found to be much lower than expected from an autosomal dominant disease. These data are inconclusive; however, they suggest that in future etiologic studies 3 types of AD should be considered separately: autosomal dominant, familial, and sporadic. Subclassification of AD by type of occurrence generates groups of patients which are probably more homogeneous regarding etiology. PMID- 3050981 TI - The importance of regressive changes in the development of the nervous system: towards a neurobiological theory of child development. AB - In this paper we propose that theories of early personality development be revised to consider knowledge of neurodevelopment. Research findings are reviewed that have established the presence of regressive changes in the normal development of the brain. These regressive changes, consisting of neuronal death and process and synapse elimination, are theoretically linked to three precursors of personality: sex differences, temperamental traits and perceptual-motor coordination. Evidence supporting these hypotheses is provided from studies examining how the effects of genetic, environmental, and experiential factors on brain development may determine the development of these personality features. PMID- 3050983 TI - The concept of cycloid psychotic disorder. AB - The concept of cycloid psychosis or cycloid psychotic disorder has been used in the European psychiatric literature for almost half a century. However, it has now for the first time been comprised into the 10th revision of the World Health Organization's International Classification of Diseases (ICD-10) that is currently in the phase of field trials. In this article evidence is presented that supports the independence of cycloid psychotic disorder from other major psychotic disorders. In particular, evidence is presented in favour of the clinical and predictive validity of the cycloid psychosis construct. Issues related to treatment are discussed and suggestions for future research are given. PMID- 3050984 TI - Abnormal hemoglobins in China. PMID- 3050985 TI - A briefing of the Peking Union Medical College Hospital. PMID- 3050986 TI - The B- to Z-DNA equilibrium in vivo is perturbed by biological processes. AB - Right-handed B and left-handed Z conformations coexist in equilibrium in portions of plasmids in Escherichia coli. The equilibria are influenced by the length of the sequences that undergo the structural transitions and are perturbed by biological processes. The composite results of three types of determinations indicate a supercoil density of -0.025 in vivo. The coexistence of alternative DNA conformations in living cells implies the potential of these structures or their transitions for important functions in genetic regulatory processes. PMID- 3050987 TI - Coordinated assembly of multisubunit proteins: oligomerization of bacterial enterotoxins in vivo and in vitro. AB - In this paper we study the assembly, in vivo and in vitro, of a family of hexameric, heat-labile enterotoxins produced by diarrheagenic bacteria. The toxins, which consist of an A subunit and five B subunits, are assembled by a highly coordinated process that ensures secretion of the holotoxin complex. We show that (i) oxidation of cysteine residues in the B subunits is a prerequisite step for in vivo formation of B-subunit pentamers, (ii) reduction of dissociated B subunits in vitro abolishes their ability to reassemble, (iii) the kinetics of B-pentamer assembly in vivo can be mimicked under defined conditions in vitro, (iv) A subunits cannot associate with fully assembled B pentamers in vitro, and (v) A subunits cause an approximately 3-fold acceleration in the rate of B subunit pentamerization in vivo, implying that A subunits play a coordinating role in the pathway of holotoxin assembly. The last finding is likely to be of general significance, since it provides a mechanism for preferentially excluding or favoring certain intermediates in the assembly of multisubunit proteins. PMID- 3050988 TI - Chemical synthesis and enzymatic activity of a 99-residue peptide with a sequence proposed for the human immunodeficiency virus protease. AB - Retroviral proteins, including those from the human immunodeficiency virus (HIV), are synthesized as polyprotein precursors that require proteolytic cleavage to yield the mature viral proteins. A 99-residue polypeptide, encoded by the 5' end of the pol gene, has been proposed as the processing protease of HIV. The chemical synthesis of the 99-residue peptide was carried out by the solid-phase method, and the isolated product was found to exhibit specific proteolytic activity upon folding under reducing conditions. Upon size-exclusion chromatography, enzymatic activity was eluted at a point consistent with a dimeric molecular size. Specificity was demonstrated by the cleavage of the natural substrate HIV gag p55 into gag p24 and gag p17, as well as cleavage of small peptide substrates representing processing sites of HIV fusion proteins. The proteolytic action of the synthetic product could be inhibited by pepstatin, an aspartic protease inhibitor. PMID- 3050989 TI - Coregulation of processing and translation: mature 5' termini of Escherichia coli 23S ribosomal RNA form in polysomes. AB - In Escherichia coli, the final maturation of rRNA occurs in precursor particles, and recent experiments have suggested that ongoing protein synthesis may somehow be required for maturation to occur. The protein synthesis requirement for the formation of the 5' terminus of 23S rRNA has been clarified in vitro by varying the substrate of the reaction. In cell extracts, pre-23S rRNA in free ribosomes was not matured, but that in polysomes was efficiently processed. The reaction occurred in polysomes without the need for an energy source or other additives required for protein synthesis. Furthermore, when polysomes were dissociated into ribosomal subunits, they were no longer substrates for maturation; but the ribosomes became substrates again when they once more were incubated in the conditions for protein synthesis. All of these results are consistent with the notion that protein synthesis serves to form a polysomal complex that is the true substrate for maturation. Ribosomes in polysomes, possibly in the form of 70S initiation complexes, may more easily adopt a conformation that facilitates maturation cleavage. As a result, the rates of ribosome formation and protein synthesis could be coregulated. PMID- 3050991 TI - Coexpression and assembly of myosin heavy chain and myosin light chain in Escherichia coli. AB - A fragment of the Dictyostelium discoideum myosin heavy chain gene representing heavy meromyosin was coexpressed in Escherichia coli with the entire essential myosin light chain from the scallop. The expressed myosin heavy chain and essential myosin light chain copurify through ammonium sulfate fractionation, anion exchange, and gel filtration chromatography. The purified complex consists of about 1 mol of light chain per mol of heavy chain. This stoichiometry, which is that of native myosin, suggests that no special eukaryotic machinery is required for coassembly of these two proteins. By coexpressing different myosin heavy chain and myosin light chain combinations, it should be possible to study various isoforms of these two proteins, which are both products of multigene families in mammals. E. coli is thus an ideal system in which to study expression and multimeric assembly of individual components of the eukaryotic contractile apparatus. PMID- 3050990 TI - Proteins from eight eukaryotic cytochrome P-450 families share a segmented region of sequence similarity. AB - Proteins from eight eukaryotic families in the cytochrome P-450 superfamily share one region of sequence similarity. This region begins 275-310 amino acids from the amino terminus of each P-450, continues for approximately 170 residues, and ends 35-50 amino acids before the carboxyl terminus. The region can be divided into four domains of sequence similarity, each possessing its own pattern of invariant, conserved, and variable amino acids. The four domains are 56, 20, 59, and 28 residues long and are connected by three shorter segments of limited sequence similarity. The number of residues in these short segments varies with the P-450 protein but ranges from 0 to 20 residues. Consensus sequences based on these similarities can be used to determine whether the sequence of an unidentified peptide resembles that expected for a P-450. Sequence similarities between proteins sometimes reflect constraints imposed by the requirements of a common function. The fourth domain of the P-450s, for example, contains an invariant cysteine that provides the axial thiolate ligand to the heme iron. Other relationships between the four domains and P-450 function can be examined by in vitro mutagenic procedures that alter the conserved amino acids or modify the distance between domains. PMID- 3050992 TI - Construction of epithelioid sheets by transfection of mouse sarcoma cells with cDNAs for chicken cell adhesion molecules. AB - Pleiomorphic mouse sarcoma S180 cells were transfected with cDNAs for the liver cell adhesion molecule (L-CAM), the neural cell adhesion molecule (N-CAM), or both CAMs. Transfected cells expressed the appropriate CAMs at their surface and those expressing L-CAM (S180L cells) changed from adjoining spindle or round shapes to a closely linked "epithelioid" sheet when grown to confluence. Cells transfected with cDNA for N-CAM (S180N cells) also expressed this CAM on the cell surfaces and bound brain vesicles containing N-CAM but showed no phenotypic change to an epithelioid state. In S180L cells and doubly transfected (S180L/N) cells, L-CAM was concentrated at regions of cell contact and was codistributed with cortical actin. In S180N cells, N-CAM was uniformly distributed on the cell surface. When S180L cells were cocultured with S180L/N cells, N-CAM was not concentrated at boundaries between the S180L and S180L/N cells but was concentrated at boundaries between pairs of S180L/N cells. Fab' fragments of anti L-CAM dissociated the epithelioid sheets of S180L or S180L/N cells into cells with shapes resembling those of untransfected cells. Cells in epithelioid sheets were polygonal in shape but, unlike cells in true epithelia, had no basement membrane or polar structure; they also lacked tight junctions and desmosomes. Ultrastructural examination showed that, in contrast to the untransfected phenotype, cells in epithelioid sheets had large increases in adherens junctions and gap junctions. Dye coupling experiments indicated that the gap junctions were functional. The frequency of expression of both kinds of junctions was sharply decreased by treatment with anti-L-CAM Fab' fragments. These experiments provide support for the precedence hypothesis, which proposes that the linkage of cells by means of CAMs is a necessary event for the extensive expression of junctional structures. PMID- 3050993 TI - Evidence for regulation of reinitiation in translational control of GCN4 mRNA. AB - Translational control of the GCN4 gene of Saccharomyces cerevisiae is mediated by four upstream open reading frames (URFs) present in the leader of GCN4 mRNA. URFs 3 and 4 efficiently repress GCN4 expression in normal growth conditions; URFs 1 and 2 are required to overcome this repression in amino acid-starved cells. lacZ fusions to URFs 3 and 4 were used to determine the translational event that is regulated at these sequences by URFs 1 and 2. URF3-lacZ, URF4-lacZ, and GCN4-lacZ fusions are affected similarly by URFs 1 and 2 when no other URFs are present in the leader: expression from all three fusions is reduced by an amount slightly greater in repressing than in derepressing conditions. These results are inconsistent with models that postulate a differential effect of URFs 1 and 2 on initiation or elongation rates at URFs 3 and 4 versus the GCN4 coding sequences. We propose that the efficiency of reinitiation at the GCN4 AUG codon after translation of URFs 3 and 4 is the translational event that is stimulated in derepressing conditions by URFs 1 and 2. PMID- 3050994 TI - 5,7-Dihydroxytryptamine identifies living dopaminergic neurons in mesencephalic cultures. AB - The autofluorescent serotonin analogue 5,7-dihydroxytryptamine (5,7-DHT) was used to identify living catecholaminergic neurons in monolayer cultures derived from the embryonic rat mesencephalon. A high correlation between 5,7-DHT accumulation and aldehyde-induced catecholamine fluorescence as well as tyrosine hydroxylase but not dopamine-beta-hydroxylase or phenylethanolamine-N-methyltransferase immunoreactivity was found. This indicates that these cells were dopamine containing neurons. Whole-cell patch recordings showed that all mesencephalic neurons had resting membrane potentials of -50 mV or greater and input resistances ranging between 200 and 700 M omega and exhibited spontaneous action potentials and postsynaptic potentials. The duration of the action potential of the dopamine-containing neurons was characteristically longer than that of the non-dopamine-containing mesencephalic cells. In some dopamine-containing neurons, repolarization of the action potential was clearly biphasic, and the slow phase of repolarization was reversibly blocked by local application of Cd2+ or Co2+. This "shoulder" in the action potential was never observed in non-dopamine containing neurons, where Cd2+ or Co2+ application was always without effect. It is concluded that 5,7-DHT can be used to identify living dopamine-containing neurons in dissociated mesencephalic cultures and these neurons express distinct electrical properties. PMID- 3050996 TI - Embryonic chicken cornea and cartilage synthesize type IX collagen molecules with different amino-terminal domains. AB - We have analyzed embryonic chicken cornea for the presence of type IX collagen mRNA and protein. Using RNA transfer blot analysis, we demonstrate that alpha 1(IX) and alpha 2(IX) mRNAs are expressed by corneal epithelial cells at the time that the primary stromal components are synthesized. The levels of the mRNAs decrease with increasing developmental age and are barely detectable at day 11 of development. In contrast, type IX collagen protein is detectable by immunofluorescence at days 5 and 6 and undetectable by day 8. Using probes specific for alpha 1(IX) and alpha 2(IX) mRNAs, we demonstrate that the size of alpha 2(IX) mRNA is the same in cornea as in chondrocytes, the major source of type IX collagen. However, the alpha 1(IX) mRNA is about 700 nucleotides shorter in the cornea than in cartilage because the corneal form of the mRNA does not contain the 5' region that encodes the non-triple-helical amino-terminal globular domain of cartilage type IX collagen. Therefore, corneal type IX collagen must lack this domain. This structural modulation of an extracellular matrix protein is likely to contribute to the functional differences between cartilage matrix and the early corneal stroma, both of which are rich in type II collagen. PMID- 3050995 TI - Differential effect of alpha-latrotoxin on exocytosis from small synaptic vesicles and from large dense-core vesicles containing calcitonin gene-related peptide at the frog neuromuscular junction. AB - The regulatory peptide called calcitonin gene-related peptide (CGRP) was detected by immunofluorescence in frog motor neurons and motor nerve terminals. In motor nerve terminals, CGRP-like immunoreactivity was found to be segregated within large dense-core vesicles. To determine whether exocytosis from acetylcholine containing small synaptic vesicles and from CGRP-containing large dense-core vesicles can be independently stimulated, nerve-muscle preparations were exposed to alpha-latrotoxin. This toxin induced complete depletion of acetylcholine containing small synaptic vesicles but did not induce a parallel depletion of CGRP-like immunoreactivity and of large dense-core vesicles. These effects were independent of the presence of extracellular Ca2+ and occurred both at room temperature and at low temperature (1-3 degrees C). These findings suggest that exocytosis from the two vesicle populations is mediated by distinct biochemical mechanisms, which might be differentially regulated by physiological stimuli. PMID- 3050997 TI - Properties of the insulin receptor ectodomain. AB - To study the properties of the extracellular insulin-binding domain of the human insulin receptor (hIR), we have expressed portions of the parent molecule in mammalian cells. Receptor cDNAs encoding the entire hIR ectodomain, the alpha subunit of the hIR alone, or a portion of the alpha subunit containing the cysteine-rich region were placed within an expression vector and in turn used to transfect CHO cells. Only cells expressing mRNA for the entire hIR ectodomain secreted hIR-related protein, suggesting that the truncated versions of this domain are unstable. The ectodomain molecules were extensively glycosylated, properly processed heterotetramers. Further, they bound insulin with an affinity similar to that of the intact hIR. In the electron microscope the secreted ectodomains appeared as discrete globular structures. After incubation with roughly equimolar quantities of insulin, the ectodomains associated to form loops or branched and folded linear macroarrays. However, these structures were not restricted to the specific ligand, insulin, since epidermal growth factor also produced the effect. Nevertheless, it seems that the receptor ectodomains can exist in two structural states. The conversion of the singular to the aggregated state may somehow be associated with transmembrane communication and activation of the biological response. PMID- 3050998 TI - Two mitochondrial matrix proteases act sequentially in the processing of mammalian matrix enzymes. AB - The imported precursors of the mammalian matrix enzymes malate dehydrogenase [(S) malate:NAD+ oxidoreductase, EC 1.1.1.37] and ornithine transcarbamylase (carbamoyl-phosphate:L-ornithine carbamoyltransferase, EC 2.1.3.3) are cleaved to their mature subunits in two steps, each catalyzed by matrix-localized processing proteases. The number and properties of these proteases are the subjects of this report. We have identified and characterized two distinct protease activities in a crude matrix fraction from rat liver: processing protease I, which cleaves these precursors to the corresponding intermediate form; and processing protease II, which cleaves the intermediate forms to mature subunits. Protease I is insensitive to chelation by EDTA and to inactivation with N-ethylmaleimide; protease II is inhibited by 5 mM EDTA and is inactivated by treatment with N ethylmaleimide. We have prepared from mitochondrial matrix an 800-fold-enriched protease I fraction free of protease II activity by using the following steps: ion exchange, hydroxyapatite, molecular sieving, and hydrophobic chromatography. Using similar procedures, we also have prepared an approximately 2000-fold enriched protease II fraction, which has a trace amount of contaminating protease I. This enriched protease II fraction has little or no cleavage activity toward mitochondrial precursors but rapidly and efficiently converts intermediate forms to mature size. Finally, we show that protease I alone is sufficient to cleave the precursor of a third nuclear-encoded mitochondrial protein subunit--the beta subunit of propionyl-CoA carboxylase [propanoyl-CoA:carbon dioxide ligase (ADP forming), EC 6.4.1.3]--to its mature size. PMID- 3050999 TI - Cloning and nucleotide sequence of the gene for dihydrolipoamide acetyltransferase from Saccharomyces cerevisiae. AB - A 537-base cDNA encoding a portion of Saccharomyces cerevisiae dihydrolipoamide acetyltransferase (acetyl-CoA:dihydrolipoamide S-acetyltransferase, EC 2.3.1.12) was isolated from a lambda gt11 yeast cDNA library by immunoscreening. This cDNA was subcloned and used as a probe to screen a lambda gt11 yeast genomic DNA library. Two overlapping clones were used to determine the complete sequence of the acetyltransferase gene. The composite sequence has an open reading frame of 1446 nucleotides encoding a presequence of 28 amino acids and a mature protein of 454 amino acids (Mr = 48,546). The deduced amino acid sequence contains the experimentally determined amino acid sequences of the amino terminus and two internal peptide fragments of the acetyltransferase. Hybridization analysis of yeast genomic DNA showed that the gene has a single copy. A 915-base segment of the acetyltransferase gene hybridized to a yeast mRNA of approximately equal to 1.6 kilobases. Analysis of the deduced amino acid sequence of the dihydrolipoamide acetyltransferase revealed a multidomain structure similar to those reported for the corresponding acetyltransferases from Escherichia coli and rat liver, and extensive sequence similarity among the three enzymes. However, the yeast enzyme contains only one lipoyl domain, in contrast to three lipoyl domains reported for the E. coli enzyme and apparently two for the rat liver enzyme. PMID- 3051000 TI - Electron transfer in a genetically modified bacterial reaction center containing a heterodimer. AB - Time-resolved optical measurements encompassing the femtoseconds to seconds time scales have been used to investigate Rhodobacter capsulatus reaction centers (RCs) in which the histidine residue at position 200 on the M polypeptide has been changed to a leucine by site-directed mutagenesis. The HisM200----Leu RC, which has a heterodimer consisting of a bacteriochlorophyll and a bacteriopheophytin, is capable of the primary photochemistry observed in wild type Rb. capsulatus RCs, but with an overall quantum yield reduced by about half. The lower yield resides in the initial electron transfer reaction and may be associated in part with substantial charge transfer character of the excited heterodimer. These and other comparisons between Rb. capsulatus wild-type and HisM200----Leu RCs have important implications for our understanding of the mechanism of electron transfer in the RC and the efficiency of the charge separation process. PMID- 3051001 TI - Alteration of the amino terminus of the mature sequence of a periplasmic protein can severely affect protein export in Escherichia coli. AB - Escherichia coli alkaline phosphatase, coded for by the phoA gene, is normally translocated across the cytoplasmic membrane into the periplasm with high efficiency. We have constructed a series of derivatives of the phoA gene that code for a wild-type signal sequence but result in altered amino acid sequences at the amino terminus of the mature alkaline phosphatase. Our results suggest that the presence of two positively charged amino acids very early in the mature sequence interferes significantly with protein export. In one case, phoA2AB, the presence of the sequence Arg-Ile-Arg at the amino terminus of alkaline phosphatase results in a 50-times reduction in the export of the protein. By using oligonucleotide-directed mutagenesis, we have constructed mutant derivatives of phoA2AB that are greatly enhanced for export. In all cases, these derivatives reduce the net positive charge in the region. Our results may explain the failure of E. coli to export a number of proteins coded for by artificial constructs and suggest a way to improve export in these cases. PMID- 3051003 TI - Identification of specific residues of human interleukin 2 that affect binding to the 70-kDa subunit (p70) of the interleukin 2 receptor. AB - Analogs of interleukin 2 containing defined amino acid substitutions and deletions were assayed for bioactivity and for competitive binding to the high affinity human interleukin 2 receptor complex and its two component subunits, a 55-kDa subunit (p55 or TAC) and a 70-kDa subunit (p70). Substitution of Asp20 or deletion of Phe124 resulted in inactive analog proteins that were unable to interact with the high-affinity p55/p70 complex or the intermediate-affinity p70 subunit of the interleukin 2 receptor. These analogs, however, retained the capacity to compete for binding to the low-affinity p55 subunit. The presence of the carboxylic acid in the side chain of Asp20 was necessary for effective binding to the p70 protein. In contrast, substitution of Trp121 and Leu17 created analogs that were inactive in the bioassay and all three binding assays. The effects of these mutations on protein conformation were assessed by circular dichroism. These results demonstrate that specific residues in the NH2 and COOH termini of interleukin 2 are crucial for its structure and activity. PMID- 3051002 TI - Thymotaxin: a thymic epithelial peptide chemotactic for T-cell precursors. AB - The embryonic thymus is seeded by invading hemopoietic precursor cells that differentiate intrathymically into T lymphocytes. We have recently reported that avian thymic epithelial cells secrete chemotactic peptides, which provoke oriented migration of hemopoietic precursor cells in vitro. The established rat thymic epithelial cell line IT-45 R1 produced a polypeptide that resolves as a single band in the region of 11 kDa on NaDodSO4/polyacrylamide gels. This molecule, which we have named thymotaxin, induced a chemotactic response in a subpopulation of hemopoietic cells from juvenile rat bone marrow. Responding cells were generated by short-term coculture of rat bone marrow hemopoietic cells with mouse bone marrow stroma in a steroid-free medium. Cells selected in a chemotactic chamber have a lymphoid or blast cell morphology. The phenotype of the responding cells is Thy-1+, CD4- [corrected] and CD8-. In contrast, CD8 T lymphocyte differentiation antigen was expressed after coculture with embryonic thymic monolayers, suggesting that the responding cells correspond to the precursors colonizing the thymus. PMID- 3051004 TI - Toward a population genetic analysis of Salmonella: genetic diversity and relationships among strains of serotypes S. choleraesuis, S. derby, S. dublin, S. enteritidis, S. heidelberg, S. infantis, S. newport, and S. typhimurium. AB - Variation in the chromosomal genomes of 1527 isolates of eight common serotypes (O and H antigen profiles) of Salmonella was assessed by analysis of electrophoretically demonstrable allelic polymorphism at 23 metabolic enzyme loci. Seventy-one distinctive electrophoretic types, representing multilocus genotypes, were identified. A basically clonal population structure was indicated by the presence of strong linkage disequilibrium among enzyme loci, the association of each serotype with a relatively small number of multilocus enzyme genotypes, and the global distribution of certain genotypes. For each of six of the serotypes, 83-96% of isolates were members of a single clone. The occurrence of each of four serotypes (S. derby, S. enteritidis, S. infantis, and S. newport) in isolates of clones belonging to several evolutionary lineages, some of which are distantly related, suggests that the horizontal transfer and recombination of chromosomal genes mediating expression of cell-surface antigens has been a significant process in the evolution of the salmonellae. Two divergent clone clusters of S. derby differ in the relative frequency with which they cause disease in birds versus mammals, and two major lineages of S. newport differ in the frequency with which their clones are associated with disease in humans versus animals. PMID- 3051007 TI - Differential autonomic control of SAN and AVN regions of the canine heart: structure and function. AB - Both anatomical and physiologic evidence for relatively rich autonomic innervation of sinoatrial (SAN) and atrioventricular (AVN) regions of the canine heart exist, with indication that SAN is especially responsive to parasympathetic, while AVN is preferentially sensitive to sympathetic regulation. The distribution of autonomic pathways are sufficiently separate and discrete that careful surgical intervention can selectively delete either parasympathetic or sympathetic nerve supplies to either (or both) SAN and AVN regions. Selective blockade by restricted injections of lidocaine (general neuronal blocker) or hexamethonium (ganglionic blocker) indicate that the vast majority (perhaps all) of vagal ganglia supplying SAN reside in the pulmonary vein fat pad and associated adipose tissues. In contrast, the vagal ganglia supplying AVN are found within a smaller fat pad overlying epicardium at the junction of inferior vena cava-inferior left atrium. These vagal pathways to either automatic cells of SAN or conductile tissues of AVN can be selectively interrupted without interfering with vagal regulation of the remaining intact system. Electroneurograms from large neurons situated within PVFP of the anesthetized, open-chest animal, reveal vigorous, phasic electrical activity associated with the cardiac and respiratory cycles, as well as with sensory stimulation of the heart, great vessels, and lungs. Spontaneous electrical activity of presently unknown origin is also observed. Direct neuronal stimulation, plus retrograde transport of fluorescent markers suggest that highly selective postganglionic intracardiac pathways may regulate discharge patterns of the sinus automatic cells. PMID- 3051005 TI - Amylin found in amyloid deposits in human type 2 diabetes mellitus may be a hormone that regulates glycogen metabolism in skeletal muscle. AB - Diabetes-associated peptide has recently been isolated and characterized from the amyloid of the islets of Langerhans in type 2 (non-insulin-dependent) diabetics, and immunoreactivity with antibodies to the peptide has been demonstrated in islet B cells of both normal and type 2 diabetic subjects. In view of the evidence presented in this paper that this 37-amino acid peptide may be a hormone present in normal individuals, we now propose the name "amylin" to replace "diabetes-associated peptide." Because increased amylin, deposited as amyloid within the islets of Langerhans, is characteristic of type 2 diabetes, the study below was performed to examine the possible effects of amylin on peripheral glucose metabolism. Whole amylin was synthesized by using solid-phase techniques, with formation of the disulfide linkage by oxidation in dilute aqueous solution and recovery of the peptide by lyophilization. The effects of amylin on glucose metabolism were studied in two preparations in vitro, isolated rat soleus muscle strips and isolated rat adipocytes. In skeletal muscle exposed to 120 nM amylin for 1 hr, there was a marked decrease in both basal and submaximally insulin stimulated rates of glycogen synthesis, which resulted in significant reduction in the rates of insulin-stimulated glucose uptake. In muscles treated with amylin there was no response at the concentration of insulin required to stimulate glucose uptake half-maximally in untreated (control) muscles. In marked contrast, amylin had no effect on either basal or insulin-stimulated rates of glucose incorporation into either CO2 or triacylglycerol in isolated adipocytes. Therefore, amylin may be a factor in the etiology of the insulin resistance in type 2 diabetes mellitus, as both deposition of the peptide in islet amyloid and decreased rates of glucose uptake and glycogen synthesis in skeletal muscle are characteristic of this condition. PMID- 3051006 TI - Divergent disease patterns in granulocyte-macrophage colony-stimulating factor transgenic mice associated with different transgene insertion sites. AB - A comparison was made of disease development in two lines of transgenic mice in which the granulocyte-macrophage colony-stimulating factor (GM-CSF) transgene was inserted in different chromosomal locations. Female-line mice (X chromosome insertion) had equivalent elevations of serum GM-CSF levels to those in male-line mice (autosomal insertion) but a shorter survival (median survival, 95 versus 145 days) and a significantly higher incidence of large inflammatory foci in skeletal muscle and gut congestion. Male-line transgenic mice had higher levels of cells in the peritoneal cavity and a higher frequency of spleen enlargement with excess erythropoiesis than female-line mice and uniquely developed fibrotic nodules in the abdominal and pleural cavities. The various diseases in GM-CSF transgenic mice are likely to have been induced by GM-CSF-stimulated products of macrophages, and in the two transgenic lines the macrophages exhibit characteristic differences in morphology and possibly functional activity. PMID- 3051008 TI - Modulation of atrioventricular conduction in vivo: integration of heart rate and autonomic neural input. PMID- 3051009 TI - Sympathetic-vagal interactions in the sinus and atrioventricular nodes. PMID- 3051011 TI - Clinical electrophysiology of the sinus node in man. PMID- 3051010 TI - On the mechanisms of cardiac electrophysiologic actions of adenosine and adenosine 5'-triphosphate. AB - Adenosine and ATP exert pronounced electrophysiologic actions on the mammalian heart including a negative chronotropic action on cardiac pacemakers and a negative dromotropic action on atrioventricular conduction. These actions are modulated by complex interactions of the two compounds with the autonomic nervous system. Since both adenosine and ATP are released from myocardial cells under physiologic and pathophysiologic conditions, they could play an important modulating role in cardiac electrophysiology. Indeed, studies during the last decade have yielded strong evidence for the role of adenosine in the genesis of specific arrhythmias associated with myocardial ischemia. Further studies are required to fully elucidate the mechanisms of actions of adenosine and ATP in vivo. These will undoubtedly enhance the understanding of the potential arrhythmogenic, as well as the antiarrhythmic actions of endogenous and exogenous adenosine and ATP. PMID- 3051012 TI - Electrophysiology and autonomic influences of the human atrioventricular node. PMID- 3051013 TI - Concealed conduction at the AV nodal level. PMID- 3051015 TI - Nonpharmacologic management of patients with supraventricular tachyarrhythmias. PMID- 3051014 TI - Morphology of the sinus and atrioventricular nodes and their innervation. PMID- 3051016 TI - The role of the sinus node in supraventricular arrhythmias. PMID- 3051017 TI - Mechanisms of pacemaker synchronization in the sinus node. PMID- 3051018 TI - Identification of endocrine responsive breast and endometrial carcinoma using steroid hormone receptors. PMID- 3051019 TI - Predictive tests for cancer chemotherapy and the problem of tumor cell heterogeneity. PMID- 3051020 TI - The 6-day subrenal capsule assay (SRCA): its criticism, biology and review of assay/clinical correlations. PMID- 3051021 TI - Development of human tumor cell line panels for use in disease-oriented drug screening. PMID- 3051022 TI - Szent-Gyorgyi and the bioflavonoids: new results and perspectives of pharmacological research into benzo-pyrone derivatives. Commemoration on the 50th anniversary of the award of the Nobel Prize. PMID- 3051024 TI - Effects of flavonoid compounds on the immune response. PMID- 3051023 TI - Anthocyanins as natural food colorants. PMID- 3051025 TI - Flavonoids in the environment: structure-activity relationships. PMID- 3051026 TI - The effect of flavonoids on blood-vessel wall interactions. PMID- 3051028 TI - Present status and prospects of flavonoids as anti-viral agents. PMID- 3051027 TI - Flavonoids: a new class of anticancer agents? Preclinical and clinical data of flavone acetic acid. PMID- 3051029 TI - Do natural compounds need specific drug development? PMID- 3051031 TI - Tannins and plant-animal interactions. PMID- 3051030 TI - Occurrence of flavonoid aglycones in medicinal plants. PMID- 3051032 TI - Mycoplasma infection of the fetus and newborn. PMID- 3051034 TI - The development of the brain and teratogenesis. PMID- 3051033 TI - Antenatal malarial infections. PMID- 3051035 TI - Maternal phenylketonuria. PMID- 3051036 TI - Developmental actions of gonadal steroids. PMID- 3051037 TI - Developmental effects of neuroactive drugs. PMID- 3051039 TI - Molecular biologic approaches to prenatal defects. PMID- 3051040 TI - Acute bacterial chorioamnionitis. PMID- 3051041 TI - Congenital syphilis. PMID- 3051038 TI - The neurotoxic, teratogenic, and behavioral teratogenic effects of lead at low dose: a paradigm for transplacental toxicants. PMID- 3051042 TI - Influence of alpha-tocopherol on prostacyclin synthesis by rat's arterial and myometrial tissues. AB - Influence of alpha-tocopherol on PGI2 synthesis by rat arterial and myometrial tissues was investigated using a rat platelet antiaggregatory bioassay. Chronic administration of alpha-tocopherol to female rats (10 mg kg-1 day-1 s.c. for 14 days) significantly increased ex-vivo PGI2 synthesis by the arterial tissue from 12.7 +/- 0.3 (control, mean +/- s.e.m) to 17.2 +/- 0.4 ng PGI2 mg-1 wet tissue and by the myometrial tissue (in proestrus) from 1.1 +/- 0.07 (control) to 1.85 +/- 0.1 ng PGI2 mg-1 wet tissue (P less than 0.05, n = 6). alpha-tocopherol (5 mg kg-1 day-1 for 14 days) did not stimulate PGI2 to any significant level. Pretreatment of male rat arterial tissue with alpha-tocopherol (0.02, 0.1 or 0.2 mM) in vitro increased PGI2 synthesis in a dose-dependent manner. At a dose of 0.2 mM it increased PGI2 synthesis from 13.70 +/- 0.70 (control) to 22.6 +/- 1.4 ng PGI2 mg-1 wet tissue (P less than 0.1, n = 6). Pre-treatment of 14-day pregnant rat myometrium with alpha-tocopherol 0.2 and 0.4 mM significantly increased PGI2 synthesis from 1.2 +/- 0.06 (control) to 1.90 +/- 0.12 and 2.1 +/- 0.1 ng PGI2 mg-1 wet tissue, respectively (P less than 0.05, n = 6). The results indicate that the ability of alpha-tocopherol to stimulate PGI2 synthesis may partly contribute towards better understanding of the mechanisms that underly the protective effect of alpha-tocopherol against experimentally induced decreases in coronary flow and intravascular coagulations in some mammals. Furthermore adequate intake of alpha-tocopherol during pregnancy may enhance uterine blood flow and ensure adequate foetal nutrition. PMID- 3051043 TI - Prostaglandin E2 gel compared to oxytocin for medically-indicated labour induction at term: a controlled clinical trial. AB - A clinical trial was carried out to compare the efficacy and tolerance of two modes of induction of labour: vaginally administered prostaglandin E2 gel vs intravenous perfusion of synthetic oxytocin. Fifty women with pregnancy at or near term in whom prompt vaginal delivery was clinically indicated were divided at random into sub-groups of 25 each. Initial dose of PGE2 gel was 1 mg followed by another application of 1 mg or 2 mg 6 hours later in case active labour stage has not been reached. Progressive oxytocin perfusion began with 1 mU/min., being increased gradually every 20 minutes until efficacious uterine dynamics were attained. Data were recorded on entry of age, parity, gestation age, cervical dilatation, Bishop score, indication for induction. Continuous materno-foetal monitoring was carried out during the induction period. Results were evaluated from induction/delivery time, type of delivery, maternal and foetal condition, and any side-effects which developed. Apart from a higher number of instrumental deliveries in the PGE2 gel series, which was not related to the induction method, there was no significant difference between any of the variables evaluated, both methods producing active labour in approximately 70% of the patients within 12 hours. The authors stress, however, the convenience, both for patients and hospital staff, of the administration of an intravaginal induction agent over a systemic therapeutic method of induction. PMID- 3051044 TI - A controlled trial of an oral bronchodilator preparation ('Franol') in asthma. AB - In view of the lack of published data on the oral bronchodilator preparation 'Franol' (120 mg theophylline, 11 mg ephedrine hydrochloride and 8 mg phenobarbitone per tablet) a double-blind study was carried out to compare the effects on lung function of a single dose of 2 'Franol' tablets, 1 'Franol' plus 1 placebo tablet, or 2 placebo tablets over a period of 8 hours in 30 asthmatic patients with reversible airways resistance (mean FEV1 1.31 l increasing to 1.71 l after 200 micrograms salbutamol inhalation). 'Franol' produced dose-dependent bronchodilation. Two tablets caused significant bronchodilation from 90 minutes to 8 hours (peak change from baseline at 2.5 hours: FEV1 20.8%, PEFR 22.3%). One tablet of 'Franol' produced significant bronchodilation only between 60 minutes and 3 hours (change from baseline at 2.5 hours: FEV1 7.5%, PEFR 7.9%). There was no change in lung function with placebo (change from baseline at 2.5 hours: FEV1 5.4%, PEFR 5.9%). Median serum theophylline levels at 2.5 hours were 6.38 micrograms/ml for 2 tablets and 3.18 micrograms/ml for 1 tablet. Median peak phenobarbitone levels were 0.5 micrograms/ml. There were no clinically relevant changes in pulse rate and blood pressure during the study and no adverse events were reported. PMID- 3051045 TI - Amnesic effect of the novel anticonvulsant MK-801. AB - MK-801, a reported N-methyl-D-aspartate (NMDA) antagonist with affinity for the phencyclidine (PCP) receptor, injected intravenously in mice before a training trial in a passive avoidance procedure, produced a similar amnesic effect to that produced by the standard amnesic agent scopolamine. Compared to vehicle-treated mice, each drug produced significant amnesia, yet the potency of MK-801 was 40 times that of scopolamine. This result with the MK-801 is consistent with previous reports that drugs which act at PCP recognition sites within the brain produce memory impairing effects in rodents. PMID- 3051046 TI - Progress in understanding the relationship between the pharmacological effects of nicotine and human tobacco dependence. AB - The present paper is intended to serve as an introduction to the series of eight papers which follow in this issue of Pharmacology Biochemistry and Behavior. A brief historical review of research that is at the root of much recent progress is provided in the present paper. In addition, we provide some data which illustrates the scope of tobacco-related research, world wide, in an effort to provide a perspective as to the vast amount of research activity that is currently in progress. Seven of the papers which follow were presented at a symposium held under the auspices of the American Society for Pharmacology and Therapeutics in 1986. Taken together, these papers are intended to provide new data and to review major areas of pharmacologic research relevant to the understanding and treatment of tobacco dependence. The topics include the behavioral and physiologic mechanisms by which the effects of nicotine are mediated, metabolic aspects of nicotine kinetics, and genetic determinants of responses to nicotine. The final paper is a discussion of the implications of these recent data for the pharmacologic treatment of tobacco dependence. PMID- 3051047 TI - Pharmacologic determinants of tobacco self-administration by humans. AB - Tobacco is a naturally occurring source of nicotine, which is a chemical of demonstrable abuse liability and dependence potential. All commonly used forms of tobacco result in the delivery of nicotine to the central nervous system (CNS), where its actions affect the probability of subsequent use. The role of nicotine as a determinant of patterns of tobacco self-administration and other tobacco associated responses has frequently been confounded by the complexity of this form of drug self-administration, since the amount of nicotine delivered to the CNS is not a simple function of the amount of tobacco consumed. The present paper is a summary of data which indicate that nicotine administration and withdrawal are determinants of tobacco ingestion. Recent data that are reviewed include those which indicate that the following effects of nicotine bear an orderly relation to the dose administered: (1) reduction of cigarette smoking, (2) production of discriminative effects, and (3) blockade of tobacco withdrawal symptoms. A secondary intent of the present paper is to describe aspects of tobacco dependence which are relevant to the appreciation of the subsequent papers appearing in this series of eight. PMID- 3051048 TI - Reinforcing effects of nicotine in humans and experimental animals responding under intermittent schedules of i.v. drug injection. AB - The self-administration paradigm is an experimental model of drug dependence in which the reinforcing properties of drugs can be directly assessed. This paradigm avoids the possible confounding influence of nonpharmacologic factors which may contribute to drug taking in the nonlaboratory environment. When animals serve as subjects, social and cultural factors unique to humans may also be eliminated as confounding influences. Most drugs of abuse are self-administered by animals and humans under such conditions. Until 1981, laboratory studies of nicotine self administration suggested that nicotine, in its own right, was only a marginally effective reinforcer. As will be shown in the present review, a study by Goldberg and his co-workers in 1981 [13] demonstrated clearly that nicotine could serve as a highly efficacious reinforcer in laboratory animals. There are several parameters which can function to substantially strengthen the behavior which leads to nicotine ingestion. These include the following: (1) intermittent availability of nicotine, (2) intermittent presentation of nicotine-paired stimuli, and (3) concurrent schedules of food reinforcement. Initial findings from a human IV nicotine self-administration study were consistent with those from the animal studies. Together these results confirm that nicotine can function to control behavior by serving as a reinforcer for animals and humans. The results also suggest that commonly used tobacco products function as ideal nicotine delivery systems for controlling behavior since they provide discrete nicotine-paired stimuli and lend themselves to intermittent nicotine delivery. PMID- 3051049 TI - Pharmacological treatment of tobacco dependence. AB - Pharmacologically based approaches for the treatment of tobacco dependence are reviewed. The rational basis for pharmacologic treatment approaches is that tobacco dependence is partially, and critically, mediated by the actions of tobacco-delivered nicotine to the central nervous system. These actions include direct reinforcing properties of nicotine itself, tolerance and physiologic dependence, possible beneficial effects of nicotine in the alleviation of anxiety and control of weight, and neurohormonal regulation which can become important to the maintenance of emotional well-being and performance at work. Insofar as tobacco abstinence leads to negative consequences, via these biobehavioral mechanisms, pharmacologic intervention should be able to assist in initial tobacco detoxification and help tobacco abstinent persons to avoid subsequent relapse. The purpose of this review is to survey some of the efforts to develop such interventions, as well as to elucidate some of the issues relevant to such development. Four distinct approaches are discussed: (1) Nicotine replacement, in which physiologic dependence is transferred to a safer and more therapeutically manageable nicotine delivering formulation; this category includes nicotine polacrilex gum; (2) Blockade therapy, in which a drug is taken that blocks the reinforcing properties of nicotine should relapse occur; (3) Nonspecific pharmacotherapy, in which the biobehaviorally mediated correlates of tobacco abstinence are treated on a symptomatic basis; (4) Deterrent therapy, in which a drug is taken prior to smoking such that any tobacco use would produce reliable aversive effects. PMID- 3051050 TI - Comparison of the density and distribution of brain D-1 and D-2 dopamine receptors in Buffalo vs. Fischer 344 rats. AB - In vitro autoradiography was used to compare the D-1 and D-2 receptor densities in brains from Buffalo (BUFF) and Fischer 344 (F344) rats. The latter strain has been proposed as a model for human attention deficit disorder with hyperactivity (ADDH). The radioligands [3H]-SCH 23390 and [3H]-sulpiride were used to selectively identify dopamine D-1 and D-2 receptors, respectively. Certain forebrain structures from F344 rats have a higher density of D-2 receptors, but similar numbers of D-1 receptors compared to BUFF rats. Scatchard analysis of D-2 binding (in caudate-putamen) revealed a Bmax of 10.52 +/- 1.62 fmol/mg tissue and Kd of 12.72 +/- 0.93 nM in F344 rats and 3.00 +/- 0.57 and 3.87 +/- 0.58, respectively in BUFF rats (n = 3). These results support the hypothesis that high D-2 levels are correlated with certain behavioral traits, e.g., high levels of spontaneous activity. A similar increase in D-2 receptors may be responsible for some of the behavioral manifestations of ADDH in children. PMID- 3051051 TI - Autacoids from membrane phospholipids. PMID- 3051052 TI - Discrimination between the pre- and post-synaptic components of PGI2 action in guinea-pig duodenum. PMID- 3051053 TI - Influence of prolonged therapy with flunarizine on glucose, insulin and C-peptide metabolism. PMID- 3051054 TI - Vasopressin release induced by intracranial injection of eledoisin is mediated by central angiotensin II. AB - Pulse intracerebroventricular injection of eledoisin, but not of substance P, markedly increases plasma vasopressin levels in the rat. Intracerebroventricular pretreatment with sarcosine1, alanine8-angiotensin II, 1 microgram/rat, completely suppresses the effect of eledoisin, suggesting that it is mediated by angiotensin release and angiotensin II receptor activation. The vasopressin releasing effect of eledoisin is neither due to peripheral haemodynamic alterations, nor to activation of the peripheral renin-angiotensin system. It is apparently related to central angiotensin release in a specific neuronal pathway subserving vasopressin release. This effect is not secondary to inhibition by tachykinins of the brain mechanisms for angiotensin-induced drinking, but is probably expression of direct activation of specific tachykinin receptors controlling vasopressin release. PMID- 3051055 TI - [Relationships between structure and activity of anthracycline antibiotics of the rhodomycin group: inhibition kinetics of DNA and RNA viruses of the double- and single-strand types]. AB - Antiviral activity from anthracycline antibiotics of rhodomycin-type was investigated by two independent methods, which determined the infectious units and antigenity of incomplete virions. The action of rhodomycin was dependent on structure, number and position of amino sugar, which is in aglycon. The viruses of double-stranded DNA and RNA viruses was stronger inhibited as single-stranded RNA viruses. The antiviral activity were found to increase in the serial order iremycin less than adriamycin less than daunomycin less than alpha-rubicin I less than beta-rhodomycin I less than violamycin BI-complex by inhibition kinetic determined from Adeno virus. PMID- 3051056 TI - [Glutethimide derivatives: inhibitors of steroid biosynthesis as tumor chemotherapy]. PMID- 3051058 TI - The Weir Mitchell rest cure. PMID- 3051057 TI - [Retard forms of insulin by matrix inclusion or complex formation]. PMID- 3051059 TI - In pursuit of platelets--the third blood element. PMID- 3051060 TI - Osteoporosis. A review. AB - This article provides physical therapists with current information about osteoporosis. I describe the epidemiology and etiology of the condition and discuss the advantages and disadvantages of measurement techniques used to detect and quantify bone mass. I outline methods of retarding or preventing osteoporosis and present management and treatment techniques that can be used with people who already have the condition. I close by summarizing current knowledge of osteoporosis and areas of research that should be explored to enhance our ability to detect, prevent, and treat osteoporosis. PMID- 3051061 TI - Effect of lateral hypothalamic lesion on brown adipose tissue of Zucker lean and obese rats. AB - Acute (10-day) lateral hypothalamic (LH) lesion induced a reduction of food intake in both lean and obese Zucker rats which averaged about 50% over the course of the first 10 days. The aphagia associated with a fall in body weight in both genotypes which was greater than their respective pair-fed controls, indicating a change in energetic efficiency. The reduced level of BAT protein, mitochondria and GDP binding observed in the obese rat was restored after LH lesion, suggesting the reestablishment of a normal sympathetic drive to the tissue. The markedly lower plasma insulin concentration in the LH lesioned obese rat is consistent with a reduction in parasympathetic activity in these animals. Food restriction in the sham lean rat reduced BAT protein content and mitochondrial GDP binding, whereas no such changes were observed in the food restricted obese rat. This demonstrates the insensitivity of the obese rat to dietary signals and may imply that LH lesion restores diet-induced BAT thermogenesis in the obese rat. PMID- 3051062 TI - Experience with the modified Putterman procedure. AB - Results using our modification of the Putterman procedure are reported in 43 eyelids. The procedure is very effective in cases of minimal to moderate eyelid ptosis (3 mm or less) and in the presence of normal levator function. The patients, following a thorough clinical evaluation, are tested using 2.5% phenylephrine eyedrop solution, which acts as an extremely useful prognostic indicator. Depending on the degree of ptosis and the response to phenylephrine, 6 to 9 mm of combined Muller's muscle and conjunctiva is resected using a specially designed clamp under local or attended local anesthesia (no tarsal plate is resected). The incision is repaired using a running 6-0 Prolene horizontal mattress technique, and the ends are brought up through the skin and tied over the tarsal plate. In the treatment of 43 eyelids, with the exception of one slight overcorrection, there were no complications encountered with this simple procedure. Excellent results can be expected in properly selected patients, and recovery time is minimal. Our success in the last 4 years with this modified procedure allows us to strongly recommend it for the correction of mild to moderate ptosis when there is an acceptable response to phenylephrine. PMID- 3051064 TI - Synthesis of surgical wounds without skin suture. PMID- 3051063 TI - Frontal sinus injuries: primary care and management of late complications. AB - An approach to treatment of frontal sinus injuries is proposed. For injuries involving the anterior wall of the sinus in which the nasofrontal duct is patent, the anterior wall is reconstituted either primarily or using a bone graft. Injuries involving the posterior wall of the frontal sinus generally necessitate a craniotomy; the frontal sinus in these situations is cranialized and the nasofrontal duct is plugged. In primary injuries of the frontal sinus in which the nasofrontal duct is badly damaged but the posterior sinus wall intact and in late mucoceles or mucopyoceles, all sinus mucosa is stripped and the sinus is packed with cancellous bone. The authors feel that cancellous bone is a better material than fat with which to pack a sinus, particularly in the face of active infection. Three cases are presented in which infected mucopyoceles draining to the skin were treated with removal of all sinus mucosa and packing with cancellous bone, with primary healing. PMID- 3051065 TI - [The history of psychiatric terminology in Japan]. PMID- 3051066 TI - [Childhood and transsexualism]. AB - The introduction covers the ground that led the author to take an interest in transsexualism. The word "transsexualism" came into being at a specific time: 1953 (Benjamin). The concept, the facts, covered by this word, have existed in several known forms, as well as in "anonymous" ones. But the possibility of changing sex by hormonal or surgical means has given a new twist to the problem, with the role of the doctor and the media that go along with it. The definition is studied carefully and gives a description of transsexuals in its present forms working with biological males and biological females. The author suggests substituting "sexuel" and "sexue" in French for the distinction made in English between "sex" and "gender". The follow up should give a crucial value in order to justify turning a healthy subject into one who lives between the two genders. Unfortunately, such studies are neither numerous nor completely satisfactory because of insurmountable hurdles: a limited number of subjects who have been followed up, the impossibility of making up a test group, etc. Over the past few years, a reaction has sprung up, giving psychotherapy a more important role in treating patients, taking advantage of the treatment borderline cases have been given and what it has taught us. In all fairness, no one can speak of transsexual or transvestite children as has been done in the past, but only of feminine or effeminate boys and tomboy girls. When samples of such children have been followed longitudinally, one realizes that an extremely small number of them becomes transsexual, becoming for the most part homo- or bi-sexual, though some become heterosexual. Treating these children and their parents seems very important to everyone, given how hard it is to treat adult transsexuals. Only a few kinds of treatment have been published, and more especially, there is no data on the long-term future of those children having received treatment. We are trying to bring together data on the childhood of adult patients. Often, they have little to say about their own childhood. Interviews with parents give still another point of view. Most of the cases seen in consultation resulting from problems with gender-identity are mixed and secondary, rarely in a pure, clear cut state.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3051067 TI - [Short-term psychoanalytic psychotherapy in children. Hypothetical model and clinical experiences in late childhood]. AB - This article presents several hypotheses for a conceptual and technical model of short-term psychotherapy applied to a child's emotional problems, which were brought up and discussed at a clinical research workshop at the Institute of Child Neuropsychiatry at the University of Rome. By bringing up the clinical sources the conceptual principals mentioned and the methodology referred to in the principles of short-term psychoanalytic psychotherapy defined by D. Malan (1963, 1976), the study faces the problem of variables and non-variables when adapting short-term methods from the adult to the child. In this perspective of clinical methodology the core of the article faces and discusses specific characteristics of short-term psychotherapy for children, among which: the clinical setting, diagnostic evaluation and the criteria used for selecting those for treatment; the nature of the transfer/counter-transfer relation; the way of interpreting centered on the preferred focus. PMID- 3051068 TI - [Psychodynamics of childhood instability]. AB - This work focuses on the Anglo-Saxon idea concerning "hyperactivity" and "hyperkinesis" and the French-language idea of "child psycho-motor instability". The author's own personal study (having two separate parts, on the one hand studying the psychic functioning of parent and their interaction with their child, and on the other, studying material gathered on the individual psychotherapy of unstable children), goes along with the French school of thought, highlighting the extent of incestuous sexual advances toward children (especially boys) in the family unit and the sexual nature (in the sense of child sexuality) of this excitement as the source of their instability, justifying a comparison between the unstable child and a Don Juan-type of instability. What comes out is epistemological thinking on Anglo-Saxon and French-language ideas, in particular criticism of the pre-suppositions in the Anglo-Saxon way of seeing things, which seems only to envisage the characterization of a syndromic range, rather than an organic etiology, this being more often implicit; distanced by the idea of psychodynamics, which predominate in the French-language studies, integrating the symptom of "psycho-motor instability" in the general "wholeness" of the child and evaluating ways of parent-child interaction. PMID- 3051069 TI - [Unstable or hyperkinetic children. A comparative study of the concepts]. AB - In this literature, the authors analyze and compare the concepts of instability and hyperkinesia. One historic chapter shows that we are dealing with two parallel bibliographies that don't cross-check. It's a question of two different ways of naming and describing the same children. A differential semiological analysis in relation to three conceptual pairs, chosen because of their key positions (agitation-excitation, motor weakness-dyspraxia, personality problems psychopathy) shows that it's sometimes impossible to clearly separate instability hyperkinesia from these other concepts, insofar as they have been blown out of proportion and deformed. Nosographic categories overlap and cross-penetrate each other, which greatly limits their interest. Finally, it seems as if instability, a vast idea originally, had been progressively dismembered to the advantage of other categories, then disappeared from french literature entirely only to be replaced by hyperkinesia. This hyperkinesia can be taken literally as a personality disorder like any other, and also covers whatever is left of "instability". But the very concept of hyperkinesia, as was the case with instability, is blurred. The idea of MBD, and currently TDA, has historically been placed as a return to a huge category encompassing disorders that instability had progressively been differentiated from, and might be an attempt to anchor this nosology more firmly in medical speech. But the amalgam of social, physiological and psychological data resolves nothing and the problem remains intact. PMID- 3051070 TI - [Evaluation of the institutional setting on psychotic adolescents. Psychoanalytic approach]. AB - Based on a statistical check of how certain teenage psychotics have changed, these changes, even mutative effects on these kinds of problems because of steps taken in the institutional setting, were examined. Within the framework of the mental-health facility, through compulsive repetition, a process implicating both patient and healer in their interrelationship and the possibility of talking about family problems are developed. In the same way, such a setting allows a patient's virtualities to come out and be used because of the objective, transitional and narcissistic aspects that these various opportunities bring into play. Several clinical examples are given, regarding dialectic articulation, with their theory highlighted. The idea of accepting one's own exaggerated narcissism turns out to be fundamental in realizing a teenager's psychotic problems as much as different diachronic and synchronic ways of action, so that, without playing with language, we can qualify the setting as therapeutic. PMID- 3051071 TI - Tyramine sulfate excretion may be a better predictor of antidepressant response than monoamine oxidase activity. AB - The tyramine sulfate excretion test was performed on 62 nonmelancholic depressed outpatients who then took part in a 6-week double-blind trial comparing imipramine, phenelzine, and placebo. In a double-blind design, nonresponders were switched to one of the active medications. Tyramine sulfate excretion failed to differentiate response from nonresponse to placebo. By contrast, phenelzine responders excreted significantly less tyramine sulfate than did phenelzine nonresponders, while there was a trend in the same direction for imipramine treated patients. The presence of only eight phenelzine nonresponders dictates caution in interpreting these results. Baseline monoamine oxidase (MAO) activity did not distinguish responders from nonresponders or correlate with tyramine sulfate excretion. Although males had significantly lower MAO activity than females, controlling for sex did not alter these negative findings. These results fail to confirm a previous report of a significant correlation between MAO activity and treatment response in older, mainly melancholic patients. PMID- 3051073 TI - Psychophysiology and psychopathology: a motivational approach. PMID- 3051074 TI - [Galen's teachings on the anatomo-physiology of the female genital system]. PMID- 3051072 TI - Social functioning in chronic depression: effect of 6 weeks of antidepressant treatment. AB - Social functioning was assessed in 189 nonmelancholically depressed outpatients. Patients were then treated for 6 weeks in a double-blind trial of phenelzine, imipramine, or placebo and functioning was reassessed. Before treatment, younger, more severely depressed, more chronically depressed patients and those with a DSM III diagnosis of major depression plus dysthymic disorder were more functionally impaired than patients without these characteristics. Chronically depressed patients who responded to treatment reported significantly improved functioning while nonresponders did not. These results suggest that for some chronically depressed patients, impaired functioning results at least partly from the Axis I mood disorder instead of being entirely attributable to Axis II character pathology. PMID- 3051075 TI - [The concept of organic memory in T. Ribot's works]. PMID- 3051076 TI - The modern synthesis and Lewontin's critique of sociobiology. PMID- 3051078 TI - [Malaria prevention and racial segregation. The medical preparation and justification of the Duala expropriation 1912-14]. PMID- 3051077 TI - Spanish agriculture and malaria in the 18th century. PMID- 3051079 TI - Aging reconsidered: strategies to promote health and prevent disease in old age. PMID- 3051081 TI - Recent advances in understanding the pathogenesis of asthma and its clinical implications. PMID- 3051082 TI - Antibodies to components of neutrophil cytoplasm: a new diagnostic tool in patients with Wegener's granulomatosis and systemic vasculitis. AB - Eleven patients with symptoms highly suggestive of Wegener's granulomatosis are described. In spite of extensive investigation, only in two patients was a firm histological diagnosis of Wegener's granulomatosis obtained, while the remaining patients were either diagnosed as having unclassifiable systemic vasculitis or had no histological diagnosis made. This sometimes resulted in diagnostic and therapeutic delay and irreversible organ damage. Antibodies to components of neutrophil cytoplasm--recently demonstrated to be specific for Wegener's granulomatosis--were detected by indirect immunofluorescence in 10 of 11 patients, and it appears likely that antibodies to components of neutrophil cytoplasm will prove to be of great value in early diagnosis. PMID- 3051080 TI - Human brucellosis in Kuwait: a prospective study of 400 cases. AB - The clinical pattern of 400 cases of brucellosis in Kuwait is presented. The disease was acute in 77 per cent, sub-acute in 12.5 per cent and chronic in 10.5 per cent of cases. Raw milk was the major source of infection. The major features on presentation, irrespective of the course of the disease, were fever, sweating, headache, rigors, arthralgia, myalgia, and low back pain. Hepatosplenomegaly was present in 41 per cent of cases and in 32 per cent neither liver nor spleen were palpable. The haematologic findings were not specific and hepatic dysfunction (shown by liver enzyme abnormalities) was common. Skeletal (26 per cent) and genital (8.5 per cent) changes and neurobrucellosis (7 per cent) were the major complications. The ELISA was the most sensitive and reliable diagnostic test especially in relation to chronic brucellosis and neurobrucellosis. ELISA allowed the determination of brucella-specific immunoglobulins (Ig)G, IgM and IgA in the CSF, and provided profiles of Ig, in sera, which were different in patients with chronic (elevated IgG and IgA) from those with acute (elevated IgM alone or IgG, IgM and IgA) brucellosis. Treatment with tetracycline, doxycycline or rifampicin gave a cure rate of over 91 per cent in acute and subacute brucellosis. Co trimoxazole was associated with a relapse rate of 50 per cent. Two drug combinations of streptomycin and tetracycline, streptomycin and rifampicin or streptomycin and doxycycline were effective, but one of five patients with chronic brucellosis relapsed. A combination of streptomycin, tetracycline and rifampicin with or without steroids was used successfully in neurobrucellosis, septicaemic shock and subacute bacterial endocarditis. PMID- 3051083 TI - Alzheimer's disease--current issues. PMID- 3051085 TI - Allergic interstitial nephritis: clinical features and pathogenesis. PMID- 3051084 TI - Impaired beta-cell responses improve when fasting blood glucose concentration is reduced in non-insulin-dependent diabetes. AB - Pancreatic beta-cell responses to a controlled intravenous glucose stimulus in 18 untreated non-insulin-dependent diabetic patients were compared with those in seven healthy control subjects. The patients' first-and second-phase responses were only 10 to 20 per cent of those of the normal subjects. However, when normal subjects had been hyperglycaemic for two hours, their first-phase responses were similar to those of the patients. Six patients were subsequently treated by diet alone, six by diet and tolbutamide and six by diet and metformin. There was no improvement in first-phase responses after treatment, but second-phase responses doubled. Improved responses were seen with all treatments, and correlated with the fall in fasting blood glucose concentration during treatment. This study supports the view that hyperglycaemia impairs beta-cell function and suggests that first-phase responses are more sensitive to hyperglycaemia than second-phase responses. PMID- 3051086 TI - Absence of pyrimidine salvage and prevention of thymineless radiosensitization in Escherichia coli thyA cells fed dihydrothymine or thymine glycol. AB - Little information is available concerning the metabolic fate of radiation induced thymine base damage products once they have been excised from DNA. The present study was an attempt to determine whether or not thymine-requiring mutants of Escherichia coli could grow on dihydrothymine (DHT) and thymine glycol (TG) by "salvaging" the altered thymines. A second test of thymine product utilization was prevention of thymineless radiosensitization. Results showed that very low growth of Thy- cells on DHT or TG could be explained by the presence of less than or equal to 1% contaminating thymine in the mixtures. Radiation dose modification factors (DMFs) for thyA cells fed DHT or TG for 3 h were 1.38 +/- 0.28 and 1.26 +/- 0.24, respectively, whereas the DMF for 3 h thymine-starved cells was 1.63 +/- 0.05. The small (approximately 25%) amelioration of thymineless radiosensitization observed in DHT- or TG-fed cells could probably be explained by contaminating thymine in the medium. Although DHT is a normal metabolite in some cells, neither DHT nor TG could be used efficiently by thymine requiring cells in the protocol presented. PMID- 3051087 TI - Hydranencephaly. PMID- 3051088 TI - Tumour markers: a review. PMID- 3051089 TI - CT pelvimetry: replacing conventional with digital. PMID- 3051090 TI - Advances in rheumatology. AB - This article reviews the major contributions to our understanding of the rheumatic diseases. These include the first definite evidence of linkage between genes controlling the immune response and the major histocompatibility complex genotype; the recognition of the types and the role of the many crystals in arthritis; the application of sophisticated immunologic techniques to the understanding of the antinuclear autoantibodies; the recognition of a new disorder--Lyme arthritis; and the contributions of joint replacement and other surgeries, and the numerous drugs and procedures now available for treatment of patients. PMID- 3051091 TI - The differential diagnosis of geodes. AB - Lytic lesions close to or at the articular margins are secondary to a wide range of etiologies, including osteoarthritis, rheumatoid arthritis, hemophilic arthropathy, and crystal deposition disease as well as tumor and tumor-like conditions. In many cases the features of these "cysts" are distinctive, and the purpose of this article is to outline the characteristic hallmarks in some of the more frequently encountered arthropathies. PMID- 3051092 TI - Early changes of rheumatoid arthritis in the hand and wrist. AB - The radiographic diagnosis of rheumatoid arthritis can be suggested long before bone and joint destruction. Soft tissue swelling at the ulnar styloid is classical, but soft tissue swelling also occurs at the PIP and MCP joints. Joint space widening, loss of the lateral fat planes of the wrist, and radial carpal narrowing can all be seen prior to bony change. The earliest bony change is loss of the cortical white line on the radial aspect of the fourth and fifth metacarpal heads. PMID- 3051093 TI - The other arthritides. Roentgenologic features of osteoarthritis, erosive osteoarthritis, ankylosing spondylitis, psoriatic arthritis, Reiter's disease, multicentric reticulohistiocytosis, and progressive systemic sclerosis. AB - Osteoarthritis may be divided into primary generalized and secondary forms. Primary generalized osteoarthritis is characterized by narrowing of cartilage, marginal osteophytes, and absence of erosions. The most common sites of involvement are the distal interphalangeal joints of the fingers and the first carpometacarpal joint. Secondary osteoarthritis also results in narrowing of cartilage in the absence of erosions, but in regions of mechanical stress. Erosive osteoarthritis affects predominantly the proximal and distal interphalangeal joints, and evolves into bony fusion in 12 to 15 per cent of cases, about the same percentage of interphalangeal bony fusion that occurs in psoriatic arthritis. Ankylosing spondylitis predominates in the axial skeleton where it eventually leads to fusion of the vertebrae and sacroiliac joints. Psoriatic arthritis combines many features of rheumatoid arthritis, in which synovial inflammation predominates, and ankylosing spondylitis, in which ligamentous inflammation predominates. The hands and feet are involved to an equal extent, and in 20 per cent of patients the disorder also involves the sacroiliac joints and spine. Reiter's disease, like psoriatic arthritis, differs from ankylosing spondylitis in its inconstant involvement of the spine and greater involvement of peripheral joints. Reiter's disease differs from psoriatic arthritis in its predominant involvement of the lower limbs, particularly the feet, with relative sparing of the hands and wrists. Multicentric reticulohistiocytosis is a rare disorder in which polyarthritis usually precedes the onset of nodular cutaneous eruptions, a fact that emphasizes the importance of early roentgenologic recognition. The interphalangeal joints are the predominant sites of involvement in the hands, but eventually all of the synovium lined joints become affected, with arthritis mutilans the end result in one third of cases. The erosions are strikingly symmetrical and well circumscribed, and accompanying osteoporosis is disproportionately mild. Progressive systemic sclerosis is characterized by atrophy and dystrophic calcifications in the soft tissues, ultimately leading to joint deformities and resorption of the terminal tufts of the phalanges. Resorption of bone occurs at other sites as well, and marginal erosions may develop in the metacarpophalangeal and interphalangeal joints of the hands. PMID- 3051094 TI - Mixed connective tissue disease. AB - MCTD is characterized by clinical features that overlap those of several of the classic rheumatic and connective tissue disorders. Although MCTD has been distinguished from them by the presence of antibodies to an RNP and by specific immunoregulatory T cell circuit abnormalities, its definitive placement among the classic disorders remains controversial. Several of its reputed differential clinical, radiographic, and prognostic features have been discussed in this article. PMID- 3051095 TI - Arthritis in children. AB - Arthritis in children may be caused by many diseases and disorders. Some processes may be aborted by early diagnosis and treatment. Others may not cause true arthritis until the adult years, with a variable latent period. The spectrum includes congenital malformations, neoplasia (benign and malignant), infection, trauma, hematologic/vascular disorders, and connective tissue diseases. In all instances, early diagnosis is important. The growing skeleton has great potential to remodel if it is given adequate time. PMID- 3051096 TI - Gouty arthritis in the adult. AB - Gouty arthritis, a crystal induced arthropathy, occurs as a result of monosodium urate precipitation in joint fluid, synovium, and soft tissues. The underlying abnormality is due to an imbalance in uric acid metabolism. Adult males are most commonly affected. Most of the affected females are postmenopausal, when the serum uric acid levels in women approach those in men. Characteristic radiographic findings include soft tissue tophi, punched out erosions with sclerotic margins, monoarticular or asymmetric polyarticular joint involvement, and relative preservation of joint space. Although the radiographic changes are quite characteristic, they occur only late in the course of the disease, after many years of clinical symptoms. PMID- 3051097 TI - Some extra-articular manifestations of arthritis and complications of therapy. A pictorial essay. AB - More than 100 years ago Lasegue wrote, "Rheumatic fever licks at the joint but bites at the heart." This truth may be expanded to include nearly every joint ailment on one hand, and nearly every organ system on the other hand. In connective tissue disorders, bleeding diasthesias and metabolic and endocrine disorders, the joint manifestations are only one part of the total picture. This article shows a few examples of cardiopulmonary, gastrointestinal, and other manifestations. A few examples of iatrogenic complications are also described. The list of the examples described in this article is far from complete. PMID- 3051098 TI - Septic arthritis. AB - Infectious arthritis is a commonly encountered clinical problem which may result from articular contamination by a wide variety of organisms. Involvement of an articulation may occur by one of four mechanisms: hematogenous spread, spread from a contiguous source of infection, direct implantation, or postoperative contamination. Distribution is typically monoarticular with a swollen, erythematous, and painful joint. The radiographic differential diagnosis includes limited rheumatoid arthritis, gout, synovial osteochondromatosis, and pigmented villonodular synovitis. In order to prevent complications, including growth disturbances, articular destruction with ankylosis, osteomyelitis, or soft tissue extension, early diagnostic arthrocentesis is important. Radiographic abnormalities, which include soft tissue swelling, joint space loss, periarticular osteopenia, and central or marginal osseous erosions, may be delayed following clinical onset of infection. Advanced imaging techniques such as scintigraphy, CT, or MRI may allow accurate diagnosis of the infectious process at an earlier stage. PMID- 3051099 TI - Occupational and post-traumatic arthritis. AB - Secondary osteoarthritis (OA) may result from a number of causes including trauma or may be related to a specific occupation or activity. In patients who present with atypical OA one should consider the possibility of such an etiology. This article discusses post-traumatic osteoarthritis and specific forms of occupational OA. PMID- 3051100 TI - The painful shoulder. AB - Pain in the shoulder arises from a wide variety of abnormalities. The most common cause of acute pain in the nontraumatized shoulder is calcific tendonitis or bursitis readily identified by plain film radiography. On the other hand, the evaluation of chronic pain usually requires some combination of arthrography, CT, ultrasonography, and MRI to identify the source of the patients' complaints with certainty. The shoulder is rarely the site of a monoarticular process and is infrequently the dominant site of abnormality in a generalized articular disease. Clues to generalized disease processes are sometimes evidenced by changes within the shoulder present on the chest film. The source of pain in the shoulder was often elusive and puzzling to both the clinician and radiologist prior to the development of techniques such as arthrography, CT, and MRI that allow the precise delineation of soft tissue abnormalities. The judicious use of these techniques has, in large measure, helped to solve the puzzle. PMID- 3051102 TI - [X-ray diagnosis of occipitocervical malformations. III. Atlas and axis malformations, segmentation disorders of the upper cervical spine]. PMID- 3051101 TI - Neuropathic bone and joint disease. AB - The pathogenesis of the neuropathic joint has been a subject of controversy for many years. Two main theories of pathophysiologic pathways have evolved: (1) the neurotraumatic, which states that the changes result from mechanical trauma and repetitive injuries to an insensitive extremity or joint and (2) the neurovascular, which states that the changes result from a neurally initiated vascular reflex that leads to hyperemia, angiogenesis, and very active bone resorption by osteoclasts. Through clinical, radiographic, and pathologic observation, it appears evident that both pathways contribute to neuropathic bone and joint disease. Initially, the alteration of sympathetic control triggers a persistent hyperemia, leading to active bone resorption. There may or may not be associated pathologic fractures and subsequent repair. This depends upon the degree of joint insensitivity and whether or not it is subjected to continued weightbearing. If so, the neurotraumatic mechanisms come into play, but only secondarily. PMID- 3051103 TI - [Study of the lymph nodes in the mandibulofacial region and the neck using ultrasonic tomography]. PMID- 3051104 TI - [Differential sonographic diagnosis of focal liver lesions]. PMID- 3051105 TI - [Limitations of computed tomographic differential diagnosis of focal liver changes]. AB - Computed tomography of the liver is essential when there are discrepancies between ultrasonography, physical examination and laboratory findings. However, sometimes the referring clinicians overestimate the true accuracy of CT. Some of its major pitfalls include the limited spatial resolution and consequent false negative results in the case of small focal lesions; further disadvantages are the inadequate contrast between a lesion and surrounding liver parenchyma, and limited specificity even in perfusion studies. When typical patterns in contrast and density of a cyst, hemangioma and focal nodular hyperplasia are present an accurate diagnosis is achieved. In atypical cases and in all other liver lesions further investigations will be necessary. PMID- 3051107 TI - [Nuclear medical differentiation of focal lesions of the liver]. AB - Radio-colloid scintigraphy, hepatobiliary scintigraphy and blood pool scanning with tagged red blood cells are well-established methods of nuclear medicine for the differential diagnosis of hepatic lesions. Hepatic hemangiomas and focal nodular hyperplasia can be differentiated in over 90% of cases. The sensitivity of single-photon emission computer tomography (SPECT) in the detection of liver masses greater than 2 cm diameter is 92%-98%. PMID- 3051106 TI - [Predictive value of magnetic resonance tomography in comparison with sonography and computed tomography in the diagnosis of focal liver lesions]. AB - Ultrasonography and computed tomography are widely used for the diagnosis of focal lesions of the liver. However, the possibilities for tumor characterization and determination of potential malignancy are limited in many cases. Evaluation by magnetic resonance imaging shows a good correlation of relaxation time with the amount of stroma as a proportion of the total cell content of a lesion. This allows differentiation for a number of lesions. Thus, the combination of ultrasonography and magnetic resonance imaging could provide the ideal diagnostic procedure in many cases. PMID- 3051108 TI - [Angiographic detection of focal changes in the liver]. AB - CT and ultrasound have completely replaced angiography in the diagnosis of focal liver lesions. However, angiography of the liver is still important for delineation of the vascular anatomy and enhancing diagnostic specificity, especially prior to hepatic surgery. CT angiography using water-soluble or oily contrast materials improves the diagnostic accuracy with regard to the number of lesions detected in patients with liver metastases or hepatoma. Both techniques reveal more lesions than do standard angiographic methods. PMID- 3051109 TI - [Detection of superior mesenteric vein thrombosis following splenectomy using real-time and Doppler sonography]. AB - Superior mesenteric vein thrombosis after splenectomy is very rare. In the case described of a patient presenting with acute abdominal pain the diagnosis was made primarily by real-time and Doppler ultrasonography. This reduced the time elapsing before it was recognized that angiography and subsequent thrombectomy were indicated. PMID- 3051110 TI - An ancient Talmudic description of the cauda equina. PMID- 3051111 TI - Recurrence of ovarian and uterine neoplasms: diagnosis with transrectal US. AB - Twenty-one patients with clinically suspected recurrence of ovarian (n = 3) or uterine (n = 18) carcinoma were examined with suprapubic ultrasound (US) and transrectal US with high-frequency linear probes. The examinations were performed 3, 6, 9, and 15 months after surgery and radiation therapy. Eight patients underwent radiation therapy before surgery and ten after surgery; three underwent only surgery. Criteria for recurrence included increased anteroposterior diameter of the vaginal cuff (greater than 2.2 cm); structural alterations or presence of a mass in the vaginal cuff; and infiltration of the rectovaginal septum, bladder, and parametria. Transrectal US findings were true positive for recurrence in nine cases, true negative in ten, and false positive in two. US findings were true positive in three cases, true negative in seven, false positive in two, and false negative in three. In six cases results from US were technically poor, and no diagnosis could be made. Transrectal US was highly sensitive in detection of pelvic recurrent carcinomas, while US had little diagnostic value. The authors believe transrectal US can replace US in the evaluation of patients at risk for recurrent pelvic neoplasm. PMID- 3051112 TI - Detection of renal masses: sensitivities and specificities of excretory urography/linear tomography, US, and CT. AB - A prospective blinded study of 201 patients was performed to determine the relative sensitivities and specificities of excretory urography/linear tomography (EU/LT) and ultrasound (US) for the diagnosis of renal parenchymal masses. Computed tomography (CT) was used as a standard. EU/LT permitted detection of 10% of CT-confirmed masses (cystic or solid) less than 1 cm, 21% of lesions greater than or equal to 1 cm but less than 2 cm, 52% of lesions greater than or equal to 2 cm but less than 3 cm, and 85% of lesions 3 cm or more in diameter. US permitted detection of 26% of CT-confirmed lesions less than 1 cm, 60% of lesions greater than or equal to 1 cm but less than 2 cm, 82% of lesions greater than or equal to 2 cm but less than 3 cm, and 85% of lesions 3 cm or more in size. The results confirm the relative insensitivity of EU/LT for masses less than 3 cm in diameter and of US for masses less than 2 cm. Further, they suggest that CT may have a role not only in evaluation of cases in which the urographic or sonographic results are questionable or positive, but also in confirmation of apparently negative urographic findings when clinical suspicion of a lesion is high. PMID- 3051114 TI - Renal allografts: prospective analysis of Doppler sonography. AB - Fifty-six consecutively transplanted renal allografts were prospectively evaluated with serial Doppler sonographic examinations. Thirty-eight episodes of transplant rejection in 32 patients (63% proved pathologically) and 24 episodes of acute tubular necrosis (ATN) in 24 patients were encountered. The Doppler spectral waveform was characterized by means of the pulsatility index (PI), systolic/diastolic ratio (SDR), diastolic/systolic ratio (SDR), diastolic/systolic ratio (DSR), and resistive index (RI). Accuracy was optimized with use of top normal values as follows: PI = 1.8, SDR = 4.0, DSR = 0.25, RI = 0.75. There were no significant differences in the indices for those patients undergoing rejection versus those with ATN. The sensitivity for predicting transplant rejection was adversely affected by the history of either ATN or a previous rejection episode in the same allograft. Comparison with concurrent radionuclide examinations revealed similar sensitivities for rejection with scintigraphy and sonography. Differentiation of ATN from rejection was more reliable with scintigraphy than with sonography. PMID- 3051113 TI - Unusual causes of increased vascular impedance in renal transplants: duplex Doppler evaluation. AB - Duplex Doppler ultrasound (US) examination of the renal vasculature has proved valuable in assessing the kidney transplant. The normal renal allograft exhibits low-impedance arterial inflow similar to that seen in the normotopic kidney. The authors and others previously reported that a high vascular impedance, defined as either a pulsatility index (PI) greater than 1.8 or a resistive index greater than 0.9, indicates acute vascular rejection (AVR). Although AVR remains the most common cause of increased PI, the authors noted ten episodes among 180 serially followed-up transplants in which abnormal waveforms were clearly not due to rejection. Four other causes of increased vascular impedance are reported, including renal vein obstruction, severe acute tubular necrosis, pyelonephritis, and extrarenal compression of the graft. These new causes only slightly decrease the specificity of high vascular impedance for rejection. Furthermore, the cause can usually be recognized from the clinical history or other US findings. PMID- 3051115 TI - Fournier gangrene: diagnosis with scrotal US. AB - Skin thickening and subcutaneous air were detected at ultrasound (US) of the scrotum in a patient with normal-appearing testicles and signs and symptoms suggestive of an acute inflammatory process, such as epidydimitis or orchitis. The patient was found to have Fournier gangrene. In more advanced cases, US can demonstrate that this skin thickening and subcutaneous air extends posteriorly to include the perineum and buttocks. Because of the high mortality of this mixed anaerobic and aerobic infection, it is important to recognize Fournier gangrene early so that the correct surgical and medical treatment can be promptly instituted. To the authors' knowledge, this is the first description of the US characteristics of Fournier gangrene. PMID- 3051116 TI - Intrahepatic bile duct and portal vein anatomy revisited. AB - Seventeen normal cadaver livers were studied to assess the anatomic relationship of bile ducts to portal veins. The common bile duct, main portal vein, and hepatic artery were cannulated and injected with air, dilute contrast medium, and mineral oil, respectively. The livers were placed in anatomic position and examined with computed tomography. In the lateral segment of the left hepatic lobe, the bile ducts were anterior to the portal vein in seven cases, posterior in seven, and tortuous (ie, both anterior and posterior) in three. In the medial segment of the left lobe, the bile ducts were anterior in four cases, posterior in four, tortuous in three, and not seen in six. In the right lobe, the bile ducts were anterior in nine cases, posterior in five, tortuous in one, and not seen in two. In the porta hepatis, the bile ducts were anterior in ten cases, posterior in one, tortuous in five, and not seen in one. Histologic findings confirmed the anterior and posterior location of the bile ducts relative to the portal veins. These findings contradict the commonly held view of intrahepatic bile ducts being anterior to the portal vein and are clinically significant for techniques such as bile duct drainage. PMID- 3051117 TI - Acute allograft rejection in liver transplant recipients: lack of correlation with loss of hepatic artery diastolic flow. AB - Eighty hepatic artery Doppler ultrasound (US) examinations performed in 49 patients after liver transplantation were retrospectively analyzed to determine if loss of diastolic flow correlated with pathologic evidence of acute allograft rejection. All 80 Doppler examinations were performed within 7 days of hepatic needle biopsy. Forty-three Doppler waveforms from 27 patients showed normal diastolic flow. Seventeen Doppler studies in 17 patients showed complete absence of diastolic flow. Review of biopsy results for each group showed no significant difference in the proportion of acute allograft rejection present (42% for the normal group and 46% for the group lacking diastolic flow). The data from 53 US and biopsy examinations performed 2 days apart in 37 patients confirmed the lack of correlation between absent hepatic artery diastolic blood flow and rejection. The authors conclude that the loss of hepatic artery diastolic flow has no apparent clinical application for the diagnosis of acute hepatic allograft rejection. PMID- 3051118 TI - Infected bilomas and hepatic artery thrombosis in infant recipients of liver transplants. Interventional radiology and medical therapy as an alternative to retransplantation. AB - Fifteen children less than 12 kg in weight underwent transplantation of the liver for biliary atresia; eight survived. Five of the eight survivors had thrombosis of the hepatic artery without portal vein thrombosis. Three of the five patients with hepatic artery thrombosis developed infected bilomas, which were drained percutaneously under ultrasonographic (US) or computed tomographic (CT) guidance. Concurrent therapy with antibiotics and hyperoxygenation resulted in resolution of these intrahepatic collections. Although it had been thought that thrombosis of the hepatic artery most often results in necrosis of the graft and requires retransplantation, the five patients in this study survived without retransplantation. Diagnosis of hepatic artery thrombosis was achieved with the use of Doppler US in four cases, CT in four cases, and angiography in two cases. Duplex Doppler US is the preferred imaging modality. PMID- 3051119 TI - DSA in acute gastrointestinal hemorrhage: clinical and in vitro studies. AB - Selective intraarterial digital subtraction angiography (DSA) was used to examine 37 patients with acute gastrointestinal (GI) tract bleeding. Conventional screen film angiography was used as an adjunct to DSA when a larger field of view was needed (five patients) and when bowel motion prevented the acquisition of adequate image quality with DSA (two patients). Conventional angiography was also performed in all cases in which there were negative DSA examinations. DSA reduced the mean examination time considerably (20% reduction overall), especially for cases involving embolization therapy (35% reduction). DSA was especially valuable in the upper GI tract, where it was used to rapidly locate and/or assist in the embolization of bleeding sites in 19 of 20 patients with positive angiograms. There were 12 true-negative DSA examinations and one false-negative examination due to the limited field of view (9 inches [22.9 cm]). Bowel and respiratory motion were not important problems in the upper GI tract. In the lower GI tract, the usefulness of DSA was severely limited by the small field of view and the misregistration artifact caused by bowel motion. In an in vitro study, DSA and conventional angiography were compared as to their ability to depict several rates of extravasation of contrast material in a model of GI bleeding. DSA tended to be more sensitive for the detection of simulated extravasation (P less than .07). PMID- 3051120 TI - Application of image-guided surface coil P-31 MR spectroscopy to human liver, heart, and kidney. AB - Localized phosphorus-31 magnetic resonance (MR) spectroscopy in humans has previously been accomplished with surface coils by means of depth-resolved surface coil spectroscopy or rotating frame experiments, in which the extent of tissue sampled critically depends on surface coil placement. The authors' goal was to modify the surface coil image-selected in vivo spectroscopy (ISIS) experiment to accomplish three-dimensional volume selection through application of selective pulses in the presence of B0 gradients. Advantages of ISIS include the ability to use proton images to define the volume of interest (VOI) and reduced dependence on exact positioning of the surface coil. However, rapid replication of the surface coil ISIS experiment can cause spectral contamination from signals originating outside the VOI. A modified version of the ISIS experiment was developed to alleviate contamination under conditions of rapid replication. Applications of localized P-31 MR spectroscopy for observation of high-energy phosphorus metabolites are presented in human liver, heart, and transplanted and normal kidney. PMID- 3051121 TI - Fine-needle aspiration biopsy with a vacuum test tube. AB - A simple, low-cost, automatic aspiration system that makes use of vacuum test tubes designed for the drawing of venous blood has been used for real-time ultrasound (US)-guided fine-needle aspiration biopsy of 13 cysts (breast), an abscess (liver), and five solid (breast, liver, thyroid) masses. Because it allows the operator to perform aspiration with one hand while holding the real time US transducer with the other, and because creation of the suction is associated with no significant displacement of the needle, this approach has allowed sampling of lesions less than 1 cm in diameter. No complications have occurred. PMID- 3051122 TI - Acute radiation effects on cutaneous microvasculature: evaluation with a laser Doppler perfusion monitor. AB - Laser Doppler perfusion monitoring is a noninvasive technique for measuring blood flow in epidermal microvasculature that makes use of the frequency shift of light reflected from red blood cells. Measurements in patients undergoing radiation therapy show increases in blood flow of ten to 25 times baseline at doses above 50 Gy, and increases are observed with doses as low as 2 Gy. Follow-up measurements show rapid decreases in flow levels after completion of therapy, but levels remain elevated even at 1 year. PMID- 3051123 TI - Imaging prostate carcinoma. PMID- 3051124 TI - Wheel-within-a-wheel patterns in hepatosplenic infections. PMID- 3051125 TI - Pharmacogenetics and its clinical implications. Part II. Oxidation polymorphism. PMID- 3051126 TI - [Growth of Clostridium botulinum in media with garlic (Allium sativum)]. AB - The effect of garlic on the growth and toxin formation of Clostridium botulinum (GT) was studied in A) crude juice obtained from a pool of cloves by i) crushing, ii) pressing out and iii) filtration, and B) minced garlic (6 to 8 pieces per clove). For both, "white" and "red" garlic varieties were used. The juice (pH 5.7 to 6.0, for different batches) was activated 30 min at 37 degrees C and diluted (log 2) in PGY broth (g%: peptone (Difco) 1.0; glucose 0.5; yeast extract (Difco) 0.5; pH 7.3). A small drop from a 18 h at 37 degrees C chopped meat medium culture of a highly toxigenic autochthonous strain (110) of C. botulinum type A, was transferred to the juice dilutions, incubating anaerobically 15d at 37 degrees C. As a control of the inhibitory effect of the juice, four microorganisms were cultured 48 h at 37 degrees C in the juice dilutions (Table 1). Clove pieces were suspended to 50% (w/v) either in PGY broth or distilled water without pH adjustment. Aliquots were heated in water bath 15 min at 100 degrees C. After seeded with the A 110 strain, duplicate tubes and their controls were incubated 15 d at 37 degrees C in aerated and anaerobic conditions (Table 2). Titers of botulinum toxin were empirically estimated by the time to death of a pair of mice injected with 0.5 ml each, via IP, observed 72 h. Results are shown in tables 1 and 2. Garlic reduces (in undiluted juice, traces or 3 to 5 DL50/ml were recorded in separate experiments) but not inhibit GT.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3051127 TI - [Characterization of experimental infection with Mycobacterium lepraemurium in 2 strains of mice]. AB - A model of experimental leprosy in two strains of mice, namely CBA/J and CBi, has been developed based on: 1) the histological examination of a granuloma in the hind foot pad 200 days after inoculation of 0.30 microliter of Mycobacterium lepraemurium (6 x 10(8) MLm/ml); 2) the assessment of T lymphocytes in the granuloma identified by the alpha-naphthyl acetate method for esterase, and c) dissemination of the infection. The histological findings in the low resistance CBA/J strain included positive acid fast bacilli vacuolated cells, without lymphocytic infiltration, scarce number of T lymphocytes and a generalized and important dissemination, similarly to the one observed in human lepromatous leprosy. The histological findings in the hind foot pad granuloma of 30-40 per cent of the medium to high resistance CBi strain, consisted of vacuolated cells and lymphocytic infiltration, a large number of T cells and a scarce dissemination, similar to the human borderline leprosy. Both strains present a different susceptibility to a unique challenge with the mycobacterium which could be useful to disentangle the immunogenetic components involved, by means of appropriate selection and crosses. Furthermore, it could be of interest to perform immunoprotection assays in CBi mice, which might have some bearing on the development of a vaccine in human leprosy. PMID- 3051129 TI - The histochemical and cytochemical localization of proteases. PMID- 3051128 TI - [Evaluation of four antigens for the detection of anti-Mycobacterium bovis antibodies by enzyme immunoassay]. AB - An enzyme-linked immunosorbent assay (ELISA) for the diagnosis of bovine tuberculosis through the detection of specific seric antibodies has recently been developed in our laboratory. In order to assess its reproducibility and select the most adequate antigen, four bovine PPDs from different sources were evaluated in parallel: PPD M. bovis strain AN5, CEPANZO standard (CPZ), PPD M. bovis strain AN5, European Economic Community standard (EEC), PPD M. bovis strain AN5, prepared from non heated bacilli, killed by phenol (P) and PPD. M. bovis BCG strain prepared at the Pasteur Institute, Paris (BCG). Sera from 22 healthy cattle from tuberculosis free area and 20 bacteriologically confirmed tuberculous animals were employed in simultaneous assays. Antibody mean and standard deviations from healthy cattle expressed as optical density (OD) values were 45 +/- 22 when CPZ was used as antigen, 24 +/- 10 with EEC, 103 +/- 56 with P and 56 +/- 20 with BCG. Mean O.D. from tuberculous cattle were 588 +/- 158, 510 +/- 234, 782 +/- 138 and 441 +/- 189 with antigens CPZ, EEC, P and BCG respectively. A close correlation was observed when results obtained with EEC and P were compared with that of CPZ (r: 0.97 and 0.94 respectively). A lower specificity was achieved when BCG was used as antigen being also lower its correlation with the results obtained with CPZ (r: 0.87). It is concluded that our ELISA would achieve similar sensitivity and specificity if CPZ, EEC and P were used as antigens. On the other hand, BCG would not be suitable for this assay. PMID- 3051130 TI - Regulatory peptides in paraganglia. PMID- 3051131 TI - Program for accelerated development of new viral vaccines. PMID- 3051132 TI - Borna disease: a persistent virus infection of the central nervous system. PMID- 3051133 TI - Use of animal models to study hepatitis B virus. PMID- 3051134 TI - Effects of estradiol-17 beta and progesterone on the synthesis of prostaglandin F2 alpha, prostaglandin E2 and prostaglandin I2 by fibroblasts from human endometrium in vitro. AB - Estradiol-17 beta increases the production of prostaglandin F2 alpha (PGF2 alpha) in long term monolayer cell cultures of the human endometrium in a dose dependent manner. Progesterone in pharmacological dosage stimulates the syntheses of PGF2 alpha and of prostaglandin E2 (PGE2). The synthesis of prostaglandin I2 (PGI2) is not influenced by sex steroids in long term monolayer cell cultures of the human endometrium. PMID- 3051135 TI - [Denial in nonpsychotic adults. A discussion of functional and dysfunctional aspects]. PMID- 3051136 TI - Additional extrarenal abnormalities seen in Tc-99m DTPA renal flow study. AB - During 99mTc-DTPA renal flow studies, extrarenal abnormalities have been found to include aortic abnormalities (aneurysm, ectasia, thrombosis, and abruptly decreased flow), splenic abnormalities (enlarged, small, or absent spleen), hepatic arterialization, and very slow circulation. In addition to the above abnormal findings, we add three more extrarenal pathologies that may be concomitantly found with renal flow study: pleural effusion(s), malignancy of the abdomen, and anemia and/or skeletal metastases. PMID- 3051138 TI - Effect of glucagon-like peptide-1 on insulin secretion. AB - The insulinotropic actions of two forms of glucagon-like peptide 1 (GLP-1) containing 31 and 37 amino acid residues on perfused rat pancreas were compared with that of gastric inhibitory polypeptide (GIP), hitherto the most potent intestinal insulinotropic polypeptide known. The smaller form, C-terminally amidated GLP-1-(7-36), strongly enhanced insulin secretion stimulated by 11.1 mM D-glucose at a concentration as low as 0.1 nM. Comparable effects of GIP and GLP 1-(1-37) on insulin secretion were observed at concentrations of 1.0 nM and 10.0 nM, respectively. At the doses tested, neither GLP-1s nor GIP had any effect on insulin secretion induced by 3.3 mM D-glucose. At a concentration of 1.0 nM, GLP 1-(7-36 amide) also enhanced insulin secretion induced by 5 mM L-arginine whereas at concentrations of up to 10.0 nM, GLP-1-(1-37) did not. The results show that the smaller form of GLP-1 is more strongly insulinotropic than GIP. These findings suggest that the smaller GLP-1 may have a physiologically more important role as a modulator of insulin release. PMID- 3051137 TI - Insulin affects ionic composition of rainbow trout erythrocytes. AB - As is the case with the anucleate mammalian erythrocyte, ionic composition in anucleate red cells of rainbow trout, Salmo gairdneri, is insulin-sensitive. By comparison with erythrocytes cultured in insulin-free medium, those exposed to insulin concentrations of 20, 200 and 2000 microU/ml for 2 and 6 h exhibited dose dependent increases in potassium and water content coupled with reductions in the levels of sodium, magnesium and chloride. These observations suggest that the membrane of this type of erythrocyte possesses insulin receptors. PMID- 3051139 TI - [Clinical comparison of 99mTc-diethylenetriaminepentaacetic acid-human serum albumin (99mTc-HSA-D) and 99mTc-human serum albumin (99mTc-HSA) for cardiac blood pool imaging]. AB - 99mTc-HSA-D has been developed as a new blood pool scanning agent. Clinical comparison of 99mTc-HSA-D and 99mTc-HSA was made in 16 cases. The activity concentration of 99mTc in blood was measured during 2 hours after the injection in five cases. 99mTc-HSA-D showed higher concentration compared to 99mTc-HSA with the passage of time. Quantitative analysis of contrast between left ventricle and septum was performed on end diastolic frames of gated images 10 minutes after the injection. There was no obvious difference between 99mTc-HSA-D and 99mTc-HSA. The subjective comparison of detectability of lesions between the two agents was performed on three directional gated images. 99mTc-HSA-D was superior to 99mTc HSA, because the images of the latter deteriorated with the passage of time. On anterior view images 1 hour after the injection, left ventricle/lung and abdominal aorta/background count ratios were greater for 99mTc-HSA-D in many cases. There was no obvious difference in liver/background and kidney/background count ratios between the two agents. Urinary excretion of 99mTc was considerably lesser for 99mTc-HSA-D. The results indicated that 99mTc-HSA-D was superior to 99mTc-HSA for cardiac blood pool imaging. PMID- 3051140 TI - Chemically induced proliferation of peroxisomes: implications for risk assessment. AB - An increasing number of beneficial and economically important drugs, industrial chemicals, and agrichemicals are being found to cause a dose-related hepatomegaly in rodent species which is associated with the proliferation of the subcellular organelle, the peroxisome. The prolonged proliferation of hepatocellular peroxisomes and the enhanced production of the normal peroxisomal metabolic byproduct, hydrogen peroxide, in these animals during chronic bioassays has been hypothesized to account for the tumorigenicity of several of these compounds, most of which lack any measurable genotoxicity in in vitro and in vivo assays. This paper briefly reviews the basic morphology and enzymology of the peroxisome and its relationship to specific pathologic changes in animals. The potential impact of the mechanism of action of peroxisome proliferators upon the design of toxicity studies and, in conjunction with interspecies sensitivity data, upon risk assessment is discussed. PMID- 3051141 TI - Species comparisons in evaluating carcinogenicity in humans. AB - Some species and strains of experimental animals have such unique mechanisms of developing cancer that the extrapolation of such bioassay results to the human situation would be fraudulent. This fraudulent extrapolation could occur both qualitatively and quantitatively. Although it will be expensive, species other than the rat, mouse, and hamster should be tested, and tested at wider dose ranges than presently used, before risk assessors will have sufficient data to make legitimate risk estimates. Both species- and strain-unique mechanisms and pharmacokinetic information must be made available to the risk assessors before their estimates can be any better than "guesstimates." As more and more data become available, it will become essential that newer techniques of visualization of the data be used in order to evaluate the weight of evidence that an animal carcinogen is or is not a human carcinogen. PMID- 3051142 TI - Review of interspecies risk comparisons. AB - Use of laboratory animal data to make quantitative predictions of the risks of toxic effects in humans assumes that a relationship exists between the potencies in animals and humans and that its parameters can be estimated adequately. Such "scaling rules" have been used to predict the risks of carcinogenicity or other effects. A survey of the literature yielded only a modest number of papers devoted to the validity of these interspecies risk extrapolations, of which approximately 25 attempt quantitative comparisons for either radiation or chemical hazards. Some authors have investigated relatively large data sets in an attempt to identify the scaling rule that provides the best correlation of risks in two or more species. Others have selected a scaling rule and investigated whether its predictions from data in laboratory species match the risks found in humans. Opinion is divided on the validity of specific extrapolation rules and the utility of animal experiments for quantitative risk assessment. Correlations exist among risk levels in various species, but many factors appear to influence toxicity that are not captured in a simple scaling rule such as dose per unit weight or per unit surface area. Although scaling rules are useful, better projections will be made if case-specific factors such as pharmacokinetics can be considered. Further careful comparisons of quantitative risk estimates are needed. PMID- 3051143 TI - Drinking water guideline for ethylene thiourea, a metabolite of ethylene bisdithiocarbamate fungicides. AB - The ethylene bisdithiocarbamate fungicides are the most heavily used pesticides in Maine. Ethylene thiourea (ETU) is a metabolite and environmental decomposition product of these compounds, is highly water soluble, and has been detected in groundwater in the state. ETU is a recognized animal carcinogen and teratogen. When administered in the diet, ETU produced a significant increase in thyroid carcinomas in rats in two studies. Two strains of mice fed ETU in the diet developed an increased incidence of hepatomas and a slight increase in lymphomas. Application of the linearized multistage model resulted in virtually safe doses (10(-5) lifetime cancer risk) of 0.25 to 1.6 micrograms/kg/day. The major teratologic effect has been the development of hydrocephalus and other CNS defects postnatally, resulting in a high mortality rate among the offspring. The NOEL for this effect was 5 mg/kg in a single oral dose. Retarded parietal ossification was observed at 5 mg/kg/day. Serious nononcogenic thyroid effects, such as goiter, decreased 131I uptake, and reduced thyroxine production, have been observed. Thyroid hyperplasia was produced at doses as low as 0.3 mg/kg/day ETU ingested in the diet. Based on protection against thyroid and/or liver tumors and alteration in thyroid function, the recommended Drinking Water Guideline for ETU is determined to be 3 ppb. This will also provide protection against developmental effects, since these occur at doses that are one to two orders of magnitude higher. PMID- 3051144 TI - Putting the eco in ecotoxicology. AB - Testing of pesticides and often other persistent chemicals increasingly involves aquatic mesocosms. Since our intention is to protect natural systems, predictions of their response based on surrogate systems are desirable. Toxicity tests at a variety of levels of biological organization are now available, ranging from single-species laboratory tests to those carried out in field enclosures of portions of natural systems. Validation is essential to determine accuracy of the predictions that will almost certainly not be identical for the wide variety of testing systems now available. Some important scientific considerations in the development of protocols are discussed. PMID- 3051145 TI - Correlations between validation and definitive study results for genotoxic compounds. AB - The use of batteries of predictive tests, perhaps in lieu of costly definitive testing, requires thoughtful identification of individual assays and definition of test batteries and tiers. Generally predictive assays are appraised in a validation study which uses a number of compounds known to be positive (or negative) in the definitive assay. A predictive test meeting specified criteria of specificity (proportion of true positives detected) and sensitivity (proportion of true negatives detected) is then eligible for use in screening compounds of unknown potency. We formally derive the definitions of prevalence (proportion of true positives in a sample of N compounds), sensitivity, and specificity, and show that these arise from different multinomial distributions for validation and screening studies. The formal statistical correlations between the predictive tier and the definitive tier are derived for both types of studies. Correlations show that a predictive assay which is eliminated in a validation study because of low sensitivity or specificity may perform satisfactorily when used for screening. Conversely, a predictive assay which performs well in a validation study may have a poor correlation with definitive assay results when used for screening. PMID- 3051147 TI - [Endocardial calcification in neonatal fibroelastosis]. PMID- 3051146 TI - [Abdominal aortic aneurysms. Comparison of magnetic resonance, ultrasound, CT x ray and angiography]. AB - Twenty-four cases of abdominal aortic aneurysm were studied by means of MR Imaging, Computed Tomography (CT), Ultrasound (US) and Angiography. MR Imaging gave detailed information on the site and extension of the aneurysm. The extent of branches involvement, the presence of thrombosis, and the adjacent structures were also demonstrated. Major limitations of angiography were its morbility, and the difficult/impossible demonstration of eventual thrombi, and of the adjacent structures. CT, although extremely valuable in emergency cases and in the detection of calcifications, provided insufficient information on the involvement of the vessels originating from the aorta. US proved useful in the screening of abdominal aortic aneurysms, but lacked both the accuracy and the reliability necessary to a complete preoperative evaluation. MR Imaging proves thus to be a good investigation technique for a complete assessment of aneurysmatic lesions. Its major limitation is its inability to detect calcifications, while its major advantages are the accurate demonstration of both blood flow and eventual thrombi, and the multiplanarity and non-invasiveness of the methodology. PMID- 3051148 TI - [Leiomyoma of the bladder. Remarks on 2 cases]. PMID- 3051150 TI - [Indications for early diagnosis of congenital luxation of the hip]. PMID- 3051149 TI - [Adrenal angioma. Angiographic diagnosis]. PMID- 3051151 TI - Crystal identification in human synovial fluids. Methods and interpretation. AB - Gout is largely solved, both from diagnostic and therapeutic standpoints. Acute gout is easily suppressed and joint destruction can be prevented and at least reversed by lowering the serum uric acid level with relatively safe and very effective drugs. But the arthritides associated with the calcium-containing crystals remain untreatable by other than symptomatic or surgical means. If we had a method or a drug to remove CPPD or BCP crystal deposits from joints, would it make any difference in the severity of the arthritis? Which of the paradigms shown in Figure 5 holds for these crystals? If joint damage directly follows crystal deposition as in gout, then crystal removal should prove prophylactic. The unusual pattern of joint degeneration associated with polyarticular CPPD crystal deposition and the initial appearance of CPPD crystals in radiographically normal cartilage favors this idea. But radiologic chondrocalcinosis appearing in knees subjected years before to meniscectomy but not in the contralateral knees suggests that crystal deposition, in these cases at least, is secondary to trauma or surgery. If degeneration of cartilage precedes crystal deposition, as it probably does in the case of BCP crystals, then crystal removal may not be particularly helpful. Dieppe and his colleagues proposed that the calcium crystals provide a positive feedback (amplification) loop. This represents the minimalistic view of their importance. The biologic consequences of the calcium crystal deposition diseases are now being explored at the molecular level. Much more data are needed before more than speculative answers to the questions posed here can be formulated. Calcium crystal deposition is more common in older persons. The degenerative and destructive arthropathies associated with them will predictably become increasingly common as our population ages. PMID- 3051152 TI - Pathology of crystal deposition diseases. AB - Pathologic examinations of joint fluid and articular tissues can be valuable in diagnosis and evaluation of pathogenesis of crystal-associated arthropathies provided that careful attention is paid to processing of tissues. Urates, calcium pyrophosphate, and apatite-like crystals are most common in human lesions and are most widely studied. PMID- 3051153 TI - Pathologic calcium phosphate deposition in model systems. AB - Pathologic calcium phosphate deposition occurs mainly in tissues exposed to highly supersaturated body fluids such as urine and saliva. In vitro models employed to study pathologic calcification include aqueous solutions and hydrogels. Results from studies on de novo formation of various biologic calcium phosphates in aqueous solutions resembling various body fluids have been rewarding. Among others, the de novo results have shown that the formation conditions for each calcium phosphate crystal phase are quite different and are pH dependent, and yet all phases including brushite can form in neutral solutions. This is an important finding because when coupled to the facts that all calcium phosphates including brushite are more soluble in acidic solutions than in neutral ones and that, except in extreme cases of hypercalciuria and/or hyperphosphaturia, body fluid Ca and Pi are not very high, it suggests that in most cases pathologic calcification occurs in alkaline or near neutral milieus. HAP is ubiquitous because it is the most insoluble calcium phosphate crystal phase in alkaline or slightly acidic fluids. In fluids close to neutrality, whitlockite is the next favored, provided that the fluid Mg:Ca ration is neither too low nor too high. Although OCP is only slightly more soluble than whitlockite in near neutral milieus, it is favored only by low fluid Ca:Pi and Mg:Ca ratios and transforms easily to HAP. Except in very acidic milieus, brushite is the most soluble biologic calcium phosphate. Its formation is favored by acidic body fluids with high Ca x Pi or by neutral or slightly alkaline ones with very low Ca:Pi ratio or high Mg:Ca ratio. PMID- 3051154 TI - The inflammatory reaction to crystals. AB - Acute gout serves as a relatively well-defined paradigm for the review of the pathogenesis of acute microcrystal-induced inflammation. Inflammatory mediator systems activated directly or indirectly by urate crystals, the temporal events in a gouty paroxysm, and the possible factors responsible for the wide clinical variability of acute gout and other forms of crystal-induced inflammation are discussed. PMID- 3051155 TI - Mechanisms of connective tissue damage by crystals containing calcium. AB - From available clinical, radiographic, and synovial fluid findings, coupled with in vivo radiolabelled crystal turnover data and in vitro experimental data, a hypothesis has been formulated relative to the pathogenesis of BCP crystal deposition diseases (Fig. 2). Synovial lining cells phagocytose BCP crystals and particulate collagens in the joint fluid. During and/or after internalization these cells are stimulated in a variety of ways: 1) protease synthesis and secretion is relentlessly stimulated, which may damage joint tissues producing clinically evident loss of collagenous tissues including cartilage, bone, and tendon, and which may release additional amounts of crystals and particulate collagens into the synovial fluid, completing a vicious cycle; 2) PGE2 production is greatly augmented; 3) DNA synthesis is stimulated as a result of increased inositol phospholipid turnover and intracellular crystal dissolution. The increased number of synovial cells also augments the total local generation of proteases and prostenoids. Mechanical factors such as trauma or joint overuse also contribute to the pathogenesis of joint destruction as discussed in the article on the clinical aspects of BCP crystal deposition. PMID- 3051156 TI - Clinical aspects of monosodium urate monohydrate crystal deposition disease (gout). AB - Gout is a clinical syndrome with a limited range of manifestations arising as a result of the deposition of crystals of monosodium urate, the final product of purine metabolism in humans. Hyperuricemia is a common chemical aberration that is most often mild and remains asymptomatic. Thus, hyperuricemia should be distinguished from gout, even though urate supersaturation is necessary for the expression of gout. Uric acid overproduction and diminished renal uric acid excretion are the major mechanisms resulting in hyperuricemia, and an understanding of the basis of hyperuricemia in individual gout patients is an important step in determining appropriate treatment and in identifying underlying disorders, offending drugs and toxins, and inherited enzyme defects, all of which can result in hyperuricemia and gout. A scheme is presented for the evaluation of patients with new-onset gout, along with a discussion of the relationships between gout/hyperuricemia and a variety of metabolic disorders that are unusually prevalent in gouty populations. PMID- 3051157 TI - Mixed crystal deposition. AB - Mixtures of crystals are found in joint tissues more commonly than would be expected by chance. In osteoarthritic joints, for example, mixtures of different calcium salts are more common than any one type of crystal alone. This is partly explained by the predisposing factors and mechanisms of crystal formation, many of which are not salt specific. It is suggested that rheumatic disorders should not be diagnosed or classified by crystal type. PMID- 3051159 TI - Therapy in gout. AB - The effective management of patients with gout is outlined. The treatment of the acute attack, the prevention of recurrent episodes, and the dissolution of tophi, when present, are generally straightforward and associated with relatively few complications. Patients with a resistant acute attack, with extensive tophaceous deposition, or with allergy or toxicity to any of the standard drugs, present more complex treatment decisions. All agents must be used in an individualized manner for each patient with appropriate concern for risks as well as for benefit. PMID- 3051158 TI - Clinical aspects of basic calcium phosphate crystal deposition. AB - Our present understanding of the mechanisms of pathologic calcification is quite limited. Therefore, no reliable method exists to prevent calcium crystal deposition. Low doses of warfarin have been employed in some cases of soft tissue calcification because it depresses the synthesis of the vitamin K-dependent GLA protein (gamma carboxyglutamic acid), which has been implicated in the process of calcification. Reports of success must be tempered by the lack of a controlled study. Probenecid has also been utilized in the treatment of calcinosis. PMID- 3051160 TI - Approaches to gene therapy in disorders of purine metabolism. AB - Model studies with gene transfer technology using retroviral vectors expressing the cloned human genes for adenosine deaminase (ADA), purine nucleoside phosphorylase (PNP), and hypoxanthine guanine phosphoribosyltransferase (HPRT) indicate that these disorders may eventually be treated by the introduction of the normal gene into defective cells. The autologous transplantation of bone marrow infected in vitro with retroviral vectors is the most developed approach at this time. The potential usefulness of this and other approaches for each of these three disorders of purine metabolism are discussed. PMID- 3051161 TI - The natural history of repaired myelomeningocele. AB - With aggressive management based on careful evaluation, children afflicted with myelomeningocele can achieve their maximum potential. Sonography, MR imaging and CT myelography all play important roles in their evaluation. PMID- 3051162 TI - Application of image enhancement techniques to magnetic resonance imaging. AB - Image enhancement techniques are widely available, but their applications are not well defined. The authors encourage radiologists to become familiar with these techniques, to evaluate them, and to incorporate them into specific display protocols. PMID- 3051163 TI - Pediatric case of the day. Infected ovarian cyst. PMID- 3051164 TI - Identification of co-worker involvement in supported employment: a review and analysis. AB - This article identifies the roles that co-workers have assumed in providing support to employees with handicaps. These roles included validating instructional strategies, collecting subjective evaluations, implementing training procedures, collecting social comparison information, and maintaining behavior in the context of actual employment. This review is based upon an existing research literature that has focused upon providing "support" to individuals with handicaps after they become employed. The purpose of this article is to draw attention to important new roles that co-workers are assuming. Specifically, this review is one of the first attempts at defining co-worker involvement. PMID- 3051165 TI - The behavior equivalence problem in within-subject treatment comparisons. AB - Within-subject comparisons of multiple treatment effects raise a variety of issues for applied researchers. They include potential nonreversibility of behaviors, practice or habituation effects resulting from repeated presentations of the same stimulus, and the possibility of multiple-treatment interference. It has recently been suggested that the use of item cohorts with equivalent behavioral difficulty addresses those problems. In order to meet the needs of researchers whose primary interest is in domestic, vocational, or other nonacademic skills, a procedure is described for estimating equivalent difficulty for different vocational preparation tasks. PMID- 3051166 TI - [Bone banks (allografts). Symposium]. PMID- 3051167 TI - [Utility of phase contrast microscopy in the diagnosis of hematuria in pediatric patients]. PMID- 3051168 TI - [Sensory neuropathy of the 5th cranial nerve secondary to collagenosis. (Review of the literature and report of 3 cases)]. PMID- 3051169 TI - [Use of thrombolytic agents in early myocardial infarction. A review in 1987]. PMID- 3051170 TI - [The liver in the next century]. PMID- 3051171 TI - [Comparative in vivo study of the biological potency of biosynthetic human and porcine insulins]. PMID- 3051172 TI - [Hyperthyroidism and asthma]. PMID- 3051174 TI - [Lymphomatous meningitis as the beginning of diffuse well-differentiated lymphocytic lymphoma]. PMID- 3051173 TI - [Bronchial asthma and hyperthyroidism. Considerations apropos of 2 cases]. PMID- 3051175 TI - [Tuberculosis and HIV infection. Review of 48 cases]. PMID- 3051177 TI - [Effects of anesthesia on the immune system]. PMID- 3051178 TI - [Change from conventional ventilation to high-frequency jet ventilation in patients with severe adult respiratory distress syndrome (ARDS)]. PMID- 3051176 TI - [Factors which affect the performance of subarachnoid block]. PMID- 3051179 TI - [Malignant hyperpyrexia caused by neuroleptics]. PMID- 3051180 TI - [Silent myocardial ischemia (I). Concept, significance and physiopathology]. PMID- 3051181 TI - [Leiomyosarcoma of the left atrium. Report of a case and review of the literature]. PMID- 3051183 TI - [Physiopathologic bases of functional intestinal disorders]. PMID- 3051182 TI - [Ventricular arrhythmias in dilated cardiomyopathy]. PMID- 3051184 TI - [Regarding the surname Madinaveitia]. PMID- 3051185 TI - [Metabolic indicators of early graft function in liver transplantation. Comparative study]. PMID- 3051186 TI - [Retrocostal-xiphoid hernia. Apropos of 6 cases and review of the Spanish literature]. PMID- 3051187 TI - [Ulcer of the hernia neck. Review of our experience]. PMID- 3051189 TI - [Pseudomyxoma peritonei]. PMID- 3051188 TI - [Intragastric penetration of an Angelchik prosthesis]. PMID- 3051190 TI - [Multiple intrahepatic pyogenic abscesses treated by echographically directed percutaneous puncture]. PMID- 3051191 TI - [Solitary splenic abscess, treated by puncture-drainage]. PMID- 3051192 TI - [Free radicals in digestive pathology]. PMID- 3051193 TI - [Ultrasonics in the diagnosis of acute appendicitis]. PMID- 3051194 TI - Effect of converting enzyme inhibition with captopril on baroreflex sensitivity. AB - Clinical and experimental data suggest that both Captopril and angiotensin II (AII) reduce baroreflex responsiveness, and the main action of this converting enzyme inhibitor (CEI) seems clear to suppress AII synthesis. The aim of this work is to investigate this striking similarity of effects. We have verified that CEI (4 mg/kg) originates tachycardia significantly lower (P less than 0.001) than that produced in response to a similar hypotension elicited by an unspecific vasodilator: sodium nitroprusside (10-45 micrograms/kg min). CEI SQ 20881 has been reported to increase plasma vasopressin concentrations (AVP); this peptide is also known to modify baroreflex responses and has a small direct negative chronotropic effect. However, our determinations of AVP do not show any difference between the control group and the group treated with Captopril (4.78 +/- 0.87 and 5.26 +/- 0.19 pg/ml respectively). On the other hand, although CEI did not modify the rapid responses of heart rate (HR) to changes of mean arterial pressure (MAP), the decrease of MAP induced by nitroprusside was higher in the group treated with Captopril than in control group; it could mean a baroreflex ability decrease to buffer the hypotension. However, AII elicited a strong impairment of both rapid responses of HR and the buffering of hypotension produced by NP, these actions being suggested as centrally mediated. These results could indicate that the suppression of peripheral AII synthesis and therefore, the lack of pre- and postjunctional sympathetic potentiation owing to this hormone, is responsible for the absence of tachycardia under Captopril treatment. PMID- 3051195 TI - [Current aspects of Goodpasture's syndrome]. AB - Goodpasture's syndrome is an auto-immune disorder with antibodies directed against alveolar and glomerular basement membrane. These antibodies are usually present in the serum but their detection is difficult, requiring a radio immunological technique which remains uncommon. The fixed antibodies are detected more easily and more constantly in the kidney tissue than in the lung; they appear on a kidney biopsy in the form of linear deposits of immunoglobulins on the basement membrane. Thus, the renal biopsy appears to be the essential requirement to establish a diagnosis of Goodpasture's syndrome, and one should not hesitate to perform a biopsy in the presence of diffuse alveolar haemorrhage without an evident aetiology and even in the absence of any biological evidence of renal disease. Alveolar lavage may be used to confirm a haemorrhage which has been purely alveolar. The treatment for respiratory and renal recovery, using immunological therapy includes corticosteroids, cytotoxic drugs and plasmapheresis. The exact methods are not yet fully defined during the course on certain clinical forms of the disease: for oliguric patients, dialysis is the single treatment; the isolated pulmonary forms have often been treated and improved by corticotherapy alone. The responsible antigen, probably unique, has been recently been identified as collagen type IV of the glomerular basement membrane. The pathogenesis of Goodpasture's syndrome remains unknown. The inhalation of hydrocarbons has been frequently noted. The dismal prognosis in the natural history of the disease and the therapeutic possibilities available make diagnosis an urgency. Currently, renal biopsy represents the most reliable and rapid mean of diagnosis. PMID- 3051196 TI - [Chronic bronchitis. Development, prevention]. AB - The adoption of an arbitrary epidemiological definition for chronic bronchitis has enabled some progress to be made in the understanding of the frequency and natural course of this disease. It is important to distinguish between chronic airflow obstruction and chronic hypersecretion of bronchial mucus. The prevalence of the disease can only be assessed in selective groups of the population and varies according to the characteristics of these groups, but is approximately 15% of men and 8% of women. There is relatively low mortality in France: 6/100,000 in 1985 and varies according to the departments, up to 38/100,000 inhabitants. These data ought to be interpreted with care and it is also important to take account of factors linked to their evolution. Longitudinal studies undertaken 20 years ago have allowed two hypotheses to be formulated to aid in a more precise understanding of the natural history of the disease: the first of these was the Dutch hypothesis which is currently undergoing a renewal of interest linked to epidemiological studies on HRB, and the English hypothesis which has the merit of emphasizing the principal risk factor in chronic bronchitis. Certain smokers are sensitive to tobacco, even though some others are not. Does the explanation of this lie in the relationship between obstructive ventilatory problems and bronchial hyper-reactivity? This association is discussed in the light of recent work as well as the relationship between bronchial hyperactivity, smoking and chronic bronchitis. Other risk factors have been studied: occupational hazards, air pollution and acute respiratory infections in childhood. But, in spite of all the work carried out to better define the risk factors and prognosis of the disease, this makes up a complex overall picture which is poorly understood and which should stimulate us to further research. The prevention of the disorder should be aimed at three levels; at the primary level (to prevent the appearance of the disorder) the only objective which may lead to a satisfactory solution in public health terms, a secondary level would be to identify those people or groups at special risk, but it is not recommended to undertake systematic examination of large populations in view of the fact that the early diagnosis of non-specific chronic pulmonary disease as well as special studies have not been shown to demonstrate the value of these procedures. The objective of prevention is to eliminate or neutralise factors linked to the disease.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3051199 TI - Differential diagnosis of pain in cancer patients. AB - The differential diagnosis of pain in cancer patients and in patients in whom cancer is suspected can be best attained using a technique of anamnesis that 1. Follows the associations of the patient 2. Accepts biological, psychological, and social factors without prejudice 3. Comprehends the symptom "pain" in its seven dimensions. The list of possible pains includes pain of neoplastic origin, pain due to cancer therapy, somatic pain of nonneoplastic origin, cancer pain increased by psychic factors, and psychogenic pain. The last category includes pain due to conversion and pain as an accompanying sign of the "flight-fight" reaction. PMID- 3051197 TI - [Hymecromone in the treatment of symptoms following surgery of the bile ducts]. PMID- 3051200 TI - Corticosteroids as antiemetics. PMID- 3051198 TI - [Comparative evaluation of the in vitro and in vivo antibacterial activity of ceftriaxone and piperacillin: determination of the inhibitory quotient in bronchial secretions]. PMID- 3051201 TI - Hypnotherapy as antiemetic treatment in cancer chemotherapy. PMID- 3051202 TI - Fighting alopecia in cancer chemotherapy. PMID- 3051205 TI - Psychotherapy in support of patients undergoing antineoplastic chemotherapy. PMID- 3051204 TI - Psychological issues in patients with hematological malignancies. PMID- 3051203 TI - Vitamins and cancer. PMID- 3051206 TI - Electrostimulation and neurosurgical measures in cancer pain. AB - Neurosurgery for cancer pain may always be considered when the pain no longer responds to conservative treatment methods or only at the cost of undesirable side-effects. Almost all these operations that can be considered for the cancer patient can be performed percutaneously, without general anaesthesia, without loss of blood, and with short hospitalization. Chronic pain has to be differentiated according to whether it is somatogenic or neurogenic. For somatogenic pain (pain without any neurological deficit), intrathecal or intraventricular administration of morphine-like substances through an implanted drug delivery system is the most attractive method. The classical neurosurgical interruption of a tract conducting pain between the periphery and the cerebral integration centers is an almost obsolete method, and percutaneous cordotomy can only be discussed when the pain is strictly unilateral and the prognosis of the disease relatively favorable. For neurogenic pain (pain with sensory disturbances) the only method which can be helpful is electrical stimulation with an implanted neuropacemaker connected to an electrode in the dorsal columns of the cord or in the sensory thalamic nucleus (depending on the location of the pain), since morphine has at best only a poor analgesic effect on deafferentation pain. PMID- 3051207 TI - Music therapy in support of cancer patients. PMID- 3051208 TI - Cancer: a family disruption. PMID- 3051209 TI - Treatment of pain in the cancer patient: the role of the nurse. PMID- 3051210 TI - Coagulation disorders associated with neoplastic disease. PMID- 3051211 TI - Hematological support in patients undergoing allogenetic bone marrow transplantation. AB - Bone marrow transplantation is impossible without effective support with blood components during the period of pancytopenia before graft function appears. We analyzed 39 patients with leukemia and three patients with severe aplastic anemia with regard to the pre- and postgrafting requirements for RBC and PLT transfusions. Overall a median of eight RBC and four PLT concentrates were necessary in all 42 patients after allogeneic BMT (ranges, 1-32 RBC and 1-11 PLT units). Requirements were identical irrespective of the underlying disease (ALL, AML, CML, SAA). Transfusion need for RBC and PLT concentrates increased in patients over 30 years old and with a major red blood group AB0 barrier between marrow donor and recipient. The presence of grade II-IV GvHD increased RBC requirements significantly, but not PLT requirements. In addition these patients were dependent on RBC transfusions for significantly longer periods. Only one patient required therapeutic granulocyte transfusions. In a CMV-negative patient with a CMV-negative marrow donor, who died of veno-occlusive disease, cytomegalovirus was transmitted inadvertently by a seropositive PLT concentrate in his final course. Our transfusion strategy included frozen deglycerolized RBC concentrates and single donor PLT concentrates, collected mainly from the marrow donor by a cell separator. All blood products were irradiated in vitro with 1500 cGy before transfusion. An optimal transfusion policy starting before BMT can contribute to successful bone marrow transplantation. PMID- 3051212 TI - Empirical antimicrobial therapy for febrile granulocytopenic cancer patients: lessons from four EORTC trials. PMID- 3051213 TI - Managing fungal and viral infection in the immunocompromised host. AB - The increasing number of opportunistic fungal infections in neutropenic cancer patients has become of major concern. Postmortem examinations have shown that more than half of the deaths attributed to infection were due to disseminated fungal infections. Candida species are the major pathogens. Unfortunately, the early diagnosis of serious fungal invasion is exceedingly difficult in these patients, since the manifestations of infection are ill-defined and the organisms can only rarely be isolated from blood cultures. Therefore, empirical antifungal therapy in febrile patients early in the course of disease appears necessary to prevent fungal superinfections and to control clinically undetected fungal invasion. Even if a fungal infection is diagnosed, treatment options are limited to a small number of drugs, and the chance of successful treatment is slim. Amphotericin B remains the most effective drug for systemic fungal infection; however, it is also the most toxic antimicrobical in use. Reduction of its toxicity and, simultaneously, improvement of its effectiveness can be achieved by incorporation into liposomes, although liposomal amphotericin B is not yet available commercially. The newer azoles, such as ketoconazole, vibunazole, and itraconazole are orally active and less toxic. Azoles have been successfully used in the treatment of superficial candidal infections; they are not very active in systemic candidiasis. Inhibitors of the cell wall biosynthesis of candida species include inhibitors of chitin and glucan biosynthesis. Echinocandins and papulacandins interfer with the glucan synthesis and show good anticandidal activity. Their therapeutic potential is currently being explored in clinical trials. Neutropenic patients and particularly bone marrow transplant (BMT) recipients are, in addition to fungal infections, at very high risk for herpesvirus infections: herpes simplex virus (HSV) 0-20 days after BMT, cytomegalovirus (CMV) 40-80 days after BMT, varicella zoster virus (VZV) 100-160 days after BMT, and Epstein-Barr virus (EBV) 14-21 days after BMT. Acyclovir and vidarabine are, at present, the only antiviral drugs available for HSV and VZV infections, acyclovir apparently being the more effective. It is also the drug of choice for prevention of HSV infections in severely immunocompromised, i.e., BMT, patients. Bromovinyldeoxyuridine seems to be suitable for systemic treatment of HSV-1 and VZV infections in these patients. Its potency in inhibiting HSV-1 and VZV replication is superior to that of ACV, and in therapeutic doses it is nontoxic to hematopoietic progenitor cells.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3051214 TI - Aspects of infections in children with cancer. AB - There are some differences between pediatric and adult cancer patients in terms of the problem of infection but it is doubtful whether such discrepancies justify different therapeutic approaches. For example, according to the results of EORTC trial IV, children seem to show lower overall (3% vs 19%) and infectious (2% vs 8%) mortality than adults, probably due to differences in underlying diseases and to a basic better physical integrity. Central intravenous catheters appear to be indispensable in the modern management of neoplastic diseases, especially in pediatric oncology. A prospective study performed in our institution on 157 Broviac catheters, has shown: 1. Of all microbiologically documented infections, occurring in children undergoing cancer chemotherapy and radiotherapy, 27% are related to the catheter 2. Gram-positive cocci are the leading pathogens (78%) of such infections 3. Catheter-related infections often occur in non-neutropenic patients 4. Such infections can be successfully treated without removing the intravenous device. The experience of the EORTC Cooperative Group and of other centers throughout the world shows that gram-positive cocci are increasingly being isolated as the cause of infections in cancer patients. This phenomenon is probably multifactorial in origin and appears to be more evident in pediatrics. For example, in our institution, during the last 5 years gram-positive organisms have caused 69% of all bacteremias. Moreover, these organisms, while responding poorly to the presently used empirical antibiotic regimens, seem not to be as aggressive as gram-negative organisms and cause lower mortality. There is therefore controversy over whether or not to include anti-gram-positive coverage in early empirical regimens. In our institution, a pilot study comparing ceftazidime + amikacin (C + A) and ceftazidime + vancomycin (C + V) seems to show an overall advantage for C + V. This advantage, however, is not statistically significant. One finding to be pointed out is that five of ten staphylococcal infections failed to respond to C + V and that in these cases a third antibiotic had to be added. Therefore, it is likely that, at least in institutions in which methicillin-resistant staphylococci are prevalent, a triple antibiotic empirical regimen will provide the best coverage, but this can result in overtreatment. However, only prospective, randomized and multicenter trials will be able to answer such questions definitely. PMID- 3051215 TI - Controlling emesis in patients receiving cancer chemotherapy. PMID- 3051216 TI - Importance of the glutathione level and the activity of the pentose phosphate pathway in cellular heat sensitivity. PMID- 3051217 TI - Potentiation of hyperthermia by lanthanum. PMID- 3051218 TI - Thermal enhancement of the cell killing effect of X-irradiation in mammalian cells in vitro and in a transplantable mouse tumor: influence of pH, thermotolerance, hypoxia, or misonidazole. PMID- 3051219 TI - The problem of defining thermal dose. PMID- 3051220 TI - Combined heat and X-ray toxicities in intestine and skin. PMID- 3051221 TI - Effects of hyperthermic treatments on malignant cells and animal tumors: introductory remarks. PMID- 3051222 TI - Membranes as targets for hyperthermic cell killing. PMID- 3051223 TI - Hyperthermia and microcirculatory effects of heat in animal tumors. PMID- 3051224 TI - The crooked path toward the objectivation of therapeutic experience. PMID- 3051225 TI - Overview of adjuvant radiotherapy for breast cancer. PMID- 3051226 TI - Treatment decisions in breast cancer. PMID- 3051227 TI - A critical assessment of approaches to improving the efficiency of cancer clinical trials. PMID- 3051228 TI - The value of small clinical trials. PMID- 3051229 TI - Measurement of quality of life in clinical trials of therapy. PMID- 3051230 TI - Statistical tools for subset analysis in clinical trials. PMID- 3051231 TI - Randomised trials: the case for science in medicine. PMID- 3051232 TI - Estimating the magnitude of benefits from adjuvant therapy. PMID- 3051233 TI - Treatment by protocol: assessment of gains and losses. PMID- 3051234 TI - Pulmonary function of nonsmoking patients with rheumatoid arthritis in the presence and absence of secondary Sjogren's syndrome, a controlled study. AB - One hundred nonsmoking patients with rheumatoid arthritis, including 63 with rheumatoid arthritis alone (RA group) and 37 who also had secondary Sjogren's syndrome (sSS group), underwent a detailed evaluation of their pulmonary function. The results were compared with those of 110 age-matched nonsmoking controls. Normal function was significantly less common in both patient groups than in the controls. A significant percentage of patients had small airways disease (SAD) which was observed with similar frequency in the control group. If this were excluded, then isolated impairment of carbon monoxide diffusing lung capacity (DLCO) was the most commonly detected significant abnormality in both patient groups. Restrictive disease was following in frequency in the RA group, but was absent in the sSS group. Obstructive disease was very uncommon in all of the patients. In general, there were no significant differences in the frequency of the various respiratory function abnormalities between patients with RA only and those with concomitant sSS, whereas between patients and controls, the only statistically significant differences were the higher frequencies of isolated DLCO impairment and restrictive disease in the RA group. PMID- 3051235 TI - Influence of lung volume on phrenic, hypoglossal and mylohyoid nerve activities. AB - In decerebrate, paralyzed cats, ventilated by a servo-respirator in accordance with phrenic nerve activity, we examined the influence of lung volume on the activities of the phrenic, hypoglossal and mylohyoid nerves. When lung inflation was briefly withheld, the durations of inspiration (TI) and expiration (TE) and the activities of all three nerves increased. The relative increase in hypoglossal activity greatly exceeded that of phrenic activity and was apparent earlier in the course of inspiration. This hypoglossal response was enhanced by hypercapnia and isocapnic hypoxia. The responses of mylohyoid activity were quite variable: withholding lung inflation augmented inspiratory activity in some cats, but expiratory discharge in others. Sustained increases in end-expiratory lung volume were induced by application of 3-4 cm H2O of positive end-expiratory pressure (PEEP). Steady-state PEEP did not influence nerve activities or the breathing pattern. Bilateral vagotomy increased TI, TE, and the activities of all three nerves. No response to withoholding lung inflation could be discerned after vagal section. The results provide further definition of the influence of vagally mediated, lung volume dependent reflexes on the control of upper airway muscles. These reflexes are well suited to relieve or prevent upper airway obstruction. PMID- 3051236 TI - [Cerebral abscesses. Current status and perspectives]. PMID- 3051238 TI - Fine-needle aspiration cytology of prostate: experience in a nonacademic practice. AB - Fine-needle aspiration cytology of the prostate is becoming increasingly accepted and used in the United States. This is a report of the experience of a nonacademic practice in which the aspirations were performed by a number of urologists, usually in the office, and were interpreted by a number of pathologists with varying degrees of experience in cytopathology. Of 187 patients, 159 had smears adequate for evaluation, and 99 had histologic/substantiating clinical follow-up for comparison. The resulting specificity was 97%, sensitivity was 91%, and the false-negative rate was 9%. These figures compare favorably with those reported in studies from academic university-based practices. PMID- 3051237 TI - Premalignant lesions of the prostate. AB - Two putative premalignant lesions of the prostate have been identified. Prostatic intraepithelial neoplasia (PIN) is characterized by proliferation and anaplasia of cells lining ducts and acini. Atypical adenomatous hyperplasia (AAH) consists of a localized proliferation of small round glands without cytologic atypia. PIN and AAH may be confused with well-differentiated carcinoma as well as florid hyperplasia, basal cell hyperplasia, transitional metaplasia, seminal vesicular epithelium, and atypia due to inflammation, infarction, and radiation. These premalignant lesions appear to have a high predictive value for carcinoma, and their presence on prostatic biopsy warrants further search for concurrent invasive adenocarcinoma. The use of strict morphologic criteria and uniform nomenclature will ensure standardization in the diagnosis of premalignant lesions of the prostate. PMID- 3051239 TI - Cardiopulmonary resuscitation in cats. PMID- 3051240 TI - Treatment priorities in cases of multiple trauma. PMID- 3051241 TI - Oxygen and ventilatory support for the critical patient. PMID- 3051242 TI - Feline cardiac emergencies. PMID- 3051243 TI - Ultrasound in emergency veterinary medicine. PMID- 3051244 TI - [Use of UAP culture medium in isolating mycobacteria from expectorations]. PMID- 3051245 TI - [Potassium sorbate, pH and aqueous activity as inhibitors of the growth of Staphylococcus aureus]. PMID- 3051246 TI - Effect of Proteus mirabilis on liver and spleen weight and the hemagglutinin response to SRBC in rats. PMID- 3051247 TI - [The effect of purified mouse GM-CSF on CFU-C colony growth in serum-free cultures]. AB - We investigated the effect of the ingredients in serum-free cultures, in which serum was completely replaced by albumin, cholesterol and transferrin, on the growth of murine granulocyte/macrophage progenitor cells (colony forming unit in culture: CFU-C) stimulated by serum-free PWM-SCM and the influences of purified granulocyte-macrophage colony stimulating factor (GM-CSF) in serum-free cultures. The results were as follows: 1) The number of CFU-C colonies reached a peak after 4 days of incubation. 2) A linear relationship was observed between the number of CFU-C colonies and the number of inoculated bone marrow cells. 3) Serum-free cultures could support CFU-C colony growth to the same degree as that in serum containing cultures. Also, no significant differences were found in the types of colonies grown in both cultures. 4) Bovine serum albumin (BSA) and cholesterol were considered to play the most important roles among the ingredients in serum free cultures. 5) Purified GM-CSF supported CFU-C colony growth in serum-free cultures and more than a half of the colonies formed were GM-colonies. These results showed the usefulness of our serum-free cultures for studying the granulopoiesis in vitro without the influences of various substances in the serum. PMID- 3051248 TI - [Intramural esophageal bronchogenic cyst. Apropos of a case report. Review of the literature]. PMID- 3051249 TI - [Myasthenia-multiple sclerosis: a possible pathogenetic relationship?]. PMID- 3051250 TI - [Primary myelodysplastic syndromes]. PMID- 3051251 TI - [Fat embolism]. PMID- 3051252 TI - [Simple suture technic in gastroduodenal perforation]. PMID- 3051253 TI - [Sclerosing cholangitis: review of the literature apropos of a case]. PMID- 3051254 TI - Psychological issues in exercise prescription. PMID- 3051256 TI - Muscle lipolysis during exercise. An update. PMID- 3051257 TI - Electromyostimulation from a clinical perspective. A review. AB - A proliferation of the research analysis and clinical use of electromyostimulation has occurred in sports medicine in the last decade. This manuscript will review the important findings from a clinical perspective. Specifically, this article will address the advantages of electromyostimulation over voluntary exercise indicating its greater effectiveness in the early period of rehabilitation when reflex inhibition is dominant. Other advantages of electromyostimulation include its: usefulness in training one component of an agonist muscle group; effectiveness during joint immobilisation; and possible role in altering specific muscle fibre types and enzymes. In order to facilitate the use of electromyostimulation a number of factors require consideration including proper selection of stimulus parameters, joint position and electrode size, type and placement. In addition, the research findings relative to the use of electromyostimulation at different stages of recovery as well as the concern of whether electromyostimulation should be used with or without voluntary activation of the muscle are addressed. There have been few clinical studies of the use of electromyostimulation after peripheral joint injury. The studies that have been performed focus primarily on quadriceps femoris rehabilitation after knee injury and these were classified according to diagnostic groups and discussed in this article. Finally, directions for future research are described with the hope that the great scientific effort displayed to date will be continued. PMID- 3051255 TI - Stress testing. Directions for the future. PMID- 3051259 TI - [Autoimmune diseases]. PMID- 3051261 TI - [Sleep disorders in depressive states. Neurophysiological similarities]. PMID- 3051260 TI - [Study of the laxative effect of Lactitol as opposed to lactulose in an open, randomized comparative study]. PMID- 3051258 TI - Cardiac rehabilitation exercise programme. Compliance and compliance-enhancing strategies. AB - Compliance-enhancing strategies in cardiac rehabilitation should be investigated only if it has been shown that the condition under consideration is an important cause of mortality and premature disability, that the intervention or therapy is effective, and that compliance with the intervention is poor. Coronary artery disease is the leading cause of death and premature disability in industrialized countries. Evidence from randomised controlled trials of supervised exercise rehabilitation after documentation of coronary artery disease suggests a reduction in fatal event rates, and an initial improvement in both exercise tolerance and psychosocial status, although these differences between experimental and control subjects are reduced over time. Poor compliance with supervised exercise programmes is a problem. This suggests that compliance enhancement with programmes of exercise rehabilitation for cardiac patients is an appropriate area of research. A number of issues recur in compliance research including the investigation of compliance-enhancing strategies in exercise rehabilitation. These relate to the specification of definition of compliance, the description of the experimental protocol or strategy, the selection and description of the sample to be studied, the randomisation of the sample, the selection of compliance measures, contamination and co-intervention, monitoring for decay, and various ethical issues. Compliance-enhancing strategies must be designed with these methodological issues in mind. These issues are discussed with specific reference to randomised controlled trials of compliance-enhancing strategies to cardiac exercise rehabilitation. PMID- 3051262 TI - [Appendix--appendicitis--appendectomy. A historical overview]. PMID- 3051263 TI - [Oliver Wendell Holmes' treatise on the 'Contagiousness of puerperal fever'. An American contribution to the prevention of puerperal fever]. PMID- 3051264 TI - [1 years' experience with Novopen and Insuject--a survey of 50 insulin-dependent diabetic patients]. PMID- 3051265 TI - [Assessment of hematuria today]. PMID- 3051266 TI - [Pathophysiology of chronic cough]. PMID- 3051267 TI - [Expectorants: an expensive, mostly ineffective therapy of chronic cough]. PMID- 3051268 TI - [Functions of a hospital clinical pharmacology service]. PMID- 3051269 TI - [The role of psychosocial factors in the treatment of patients with rheumatoid arthritis]. PMID- 3051270 TI - [The plasma prostaglandin level in rheumatoid arthritis in children and its dynamics as affected by anti-inflammatory therapy]. PMID- 3051271 TI - [Medical disability expertise in systemic scleroderma]. PMID- 3051272 TI - [Clinical laboratory and radioisotope characteristics of kidney involvement in systemic lupus erythematosus]. PMID- 3051273 TI - [Blood serum glycosaminoglycans in assessing the degree of activity in systemic lupus erythematosus]. PMID- 3051274 TI - [Evolution of the morphological manifestations of synovitis in patients in the initial stage of rheumatoid arthritis with different variants of its further course]. PMID- 3051275 TI - [Cardiac and aortic lesions in spondylarthritis ankylopoietica]. PMID- 3051276 TI - [Hemodynamic changes during captopril inhibition of the renin-angiotensin aldosterone system in patients with circulatory failure following heart valve prosthesis]. PMID- 3051277 TI - [Proceedings of the international symposium, "Risk factors in rheumatic diseases". Pitsynda, 17-23 November 1986]. PMID- 3051278 TI - [Horton's disease]. PMID- 3051279 TI - [The role of neutrophils in the pathogenesis of rheumatoid arthritis and their changes as affected by anti-rheumatic agents]. PMID- 3051280 TI - [Experience with the use of reflexotherapy in arthrology]. PMID- 3051281 TI - [Connective tissue metabolism in systemic lupus erythematosus, systemic scleroderma and rheumatoid arthritis]. PMID- 3051282 TI - [The inflammatory response. Definitions; the evolution of a conceptual model]. PMID- 3051284 TI - Wedge resection in rhinosurgery: a review of the literature and long-term results in a hundred cases. AB - First a historical review is given of the development of the concept of wedge resection of the bony nasal pyramid since Joseph, 1907. In a second part the technical details about wedge resection are described for different types of nasal deformities. In a third part long-term results in 100 patients, 18 months after septorhinoplasty are presented. In 61 patients unilateral and in 39 patients bilateral wedge resections were performed as one step of rhinoplasty. The long-term results were good in 93 patients, there was undercorrection in six cases, whereas one patient had an overcorrected nose. These results are better than the long-term results following classic osteotomies alone. Thus the wedge resection, for the indications as defined in this study, is a very useful step in the concept of corrective and aesthetic septorhinoplasty. PMID- 3051283 TI - Effects of D-penicillamine on mononuclear cells in vitro. AB - D-penicillamine (D-pen) inhibited pokeweed mitogen-induced plaque-forming cell (PFC) response in a dose-dependent manner. This inhibition was irreversible as preincubation for a few hours with the drug followed by washes still caused suppression of the PFC response. Pretreatment of the different mononuclear cell populations with D-pen for short periods (2-24 h) showed that both macrophages (Mo) and B lymphocytes were affected by the drug. By contrast T cells were resistant. Mo appears to be more susceptible to D-pen than B cells, and in the case of drug-treated Mo, the response was restored completely with the addition of 20% fresh Mo. Our results show that D-pen, without exogenous Cu2+, inhibits the polyclonal immunoglobulin secretion by human mononuclear cells in vitro due to a strong effect on both Mo and B cells. This may explain the decrease in serum immunoglobulin levels seen in patients with rheumatoid arthritis undergoing this therapy. PMID- 3051285 TI - A modern concept of cerebrospinal fluid diagnosis in oto- and rhinorrhea. AB - Three successive CSF investigations make it possible to identify even the smallest amount of cerebrospinal fluid (CSF) in cases of otorrhea and rhinorrhea: 1. Immunological identification of beta 2-transferrin. 2. Laboratory fluorescein identification. 3. Endoscopic fluorescein detection. As a screening procedure the beta 2-transferrin identification method is always used as the first step towards clarifying a suspect liquorrhea. In addition both fluorescein tests are used for the diagnosis depending on the result of the beta 2-transferrin identification and further measures. As a result of recent practical experience special attention is paid to the test analyses; the various possibilities of taking samples as well as mailing them. A newly developed diagnostic plan of procedure should (by using practical examples) underline the clinical significance. This study describes the most up-to-date level in CSF diagnosis and demonstrates that, when combined with a corresponding X-ray investigation, a much more exact range of indication for the surgical treatment of fractures of the base of the skull and CSF leaks is possible. PMID- 3051286 TI - [Spinal muscular atrophy]. PMID- 3051287 TI - [Degenerative neuropathies]. PMID- 3051288 TI - [Inflammatory neuropathies]. PMID- 3051289 TI - [Toxico-metabolic neuropathies]. PMID- 3051290 TI - [Muscle biopsy in neurogenic lesions]. PMID- 3051291 TI - Persistence of microfilaremia in bancroftian filariasis after diethylcarbamazine citrate therapy. AB - Of 58 patients in Leogane, Haiti, infected with Wuchereria bancrofti and treated with diethylcarbamazine citrate (DEC-C) at 6 mg/kg per day for 12 days (= 72 mg/kg), 38 (66%) of 58 continued to harbor low numbers of circulating microfilariae (median microfilariae could be demonstrated in 7 (37%) of 19 patients, with a median microfilarial density of 8 mf/ml. Three patients who continued to have circulating microfilariae after two courses of DEC-C were treated a third time. Two (67%) of the three remained microfilaria positive, both with 1 mf/ml. The results of this study clearly indicate that a high percentage of persons infected with W. bancrofti and treated with one or multiple courses of DEC-C may continue to have circulating microfilariae after treatment. We suspect that these low-level reservoir carriers substantially contribute to the transmission of filariasis and may well account for the resurgence of infection levels following control efforts. PMID- 3051292 TI - [Emergence of chloroquine-resistant malaria in West Africa: the case of Sokode (Togo)]. AB - Within the framework of its surveillance of Plasmodium falciparum chloroquine sensitivity in eight West African countries (Benin, Burkina Faso, Cote d'lvoire, Mali, Mauritany, Niger, Senegal, and Togo) the Reference Centre for Chemoresistant Malaria (CRCP) at the Organization for Coordination and Cooperation to Control Major Endemic Diseases (O.C.C.G.E.) conducted an in vivo survey in February, 1987, in Sokode (Togo). Two groups of 67 children, aged 2 to 9, received, for the first group a single 10 mg/kg dose of chloroquine; for the second group a 3-day 25 mg/kg dose, according to the WHO methodology. Thick and thin blood smears were examined on D0, D2, D3 when necessary, D4 and D7. Within the 23 children who received the 10 mg/kg dose, seven (30.4%) presented a "resistance", of which six were early RI type and 1 was RII type. Out of 44 children who received the standard dose of 25 mg/kg, two (4.6%) were resistant (early RI type resistance). These data show for the first time the appearance of in vivo chloroquine resistance in this country, and call for a withdrawal of the 10 mg/kg dose of chloroquine in the treatment of fever attacks to the benefit of a 25 mg/kg dose. Thorough studies, using in vivo and in vitro techniques, should be undertaken as soon as possible, not only in Togo but in other West African countries too, to take the exact measure of the issue. PMID- 3051295 TI - [Typical sonographic findings in hemihepatectomized or partially hepatectomized patients]. AB - The frequency of postoperative complications following immediately and at intervals after hepatic surgery, is evaluated by ultrasonography in 29 patients. In addition, the following sonographic findings--not mentioned upto now in literature--are pointed out: Enlargement of the typical cross-section of the vena cava and especially pseudotumorous vault of the parenchyma of the liver versus lumen of the vena cava; the latter may produce difficulties in discriminating local relapses in paracaval liver segments. Sclerosis of great liver veins with wall-like echoes in portal veins. After right hemihepatectomy, the portal vein could be identified by ultrasonography in only 50% of the cases. Transient oedema of the gallbladder wall and periportal fields. Tortuosity of intrahepatic vessels in the immediate postoperative course; especially after resection of great tumours of more than 7.5 cm diameter. PMID- 3051294 TI - [Digital subtraction angiography in traumatology--visualization of an arteriovenous fistula in the neck]. AB - By means of an intraarterial digital subtraction angiography a fistula due to a trauma could be visualised between the right vertebral artery and the jugular vein. The investigation was performed one year after the car accident. Visualisation with exact localisation of the beginning of the fistula at the arterial site should make it possible to perform an embolisation without affecting the vertebral artery. The clinical condition will be decisive. PMID- 3051296 TI - Ultrasonographic appearance of a simple ureterocele following surgical treatment. AB - A case history is presented, showing the ultrasonographic appearance of a simple ureterocele following surgery. The cystic appearance of the presurgical ureterocele and the gradual changes after surgery are demonstrated. Follow-up examination 17 months after surgical treatment revealed a solid tumour-like ultrasonographic image. Differential diagnosis to neoplasma of the urinary bladder is emphasized. PMID- 3051293 TI - Screening of drugs for rapid activity against Trypanosoma cruzi trypomastigotes in vitro. AB - Previous studies to find drugs with an existing product licence which were active at 4 degrees C within 24 hours and which would be suitable to prevent the transmission of Chagas' disease during blood transfusion were unsuccessful. As part of an alternative approach to identify drugs active at 37 degrees C or 25 degrees C within 2 hours, over 280 compounds were screened against bloodstream trypomastigotes of Trypanosoma cruzi Sonya strain in a microslide test in vitro. Although compounds from a wide range of chemical groups were tested only three polyene antibiotics showed outstanding trypanocidal activity. Amphotericin B, candicidin and trichomycin lysed all trypomastigotes at 8 X 10(-6) as determined by microscopical techniques. However, these compounds also showed toxicity to mammalian erythrocytes at the same concentration after a 24 hours incubation period. It seems unlikely that this approach will yield a candidate replacement for gentian violet. PMID- 3051297 TI - [Fractures of the clavicle: classification, diagnosis, therapy]. AB - Clavicular fracture is one of the most frequent skeletal lesions. In most cases the median third of the clavicula is affected (this is due to the peculiar biomechanical structure). Accompanying lesions and complications of clavicular fractures are rare. A total of 13 x-ray diagnostic techniques are described for clavicular fractures. X-ray film should, as a matter of principle, always be taken in two planes. Definitely the major part of clavicular fractures are treated conservatively (rucksack dressing), whereas surgery is reserved for few and strictly defined indications. PMID- 3051298 TI - [Drug-resistant malaria in France]. PMID- 3051299 TI - [Malaria. Biologic diagnosis. Present status and the future]. PMID- 3051301 TI - [Pernicious malarial attacks]. PMID- 3051300 TI - [Malaria. Current status of chemotherapy]. PMID- 3051303 TI - [The prevention of malaria]. PMID- 3051304 TI - [A half century of hepatology (2). An interview with Andre Paraf (by Y. Le Mercier)]. PMID- 3051302 TI - [Vaccination against malaria: current status and perspectives]. PMID- 3051305 TI - [Coronary artery spasm]. PMID- 3051306 TI - [Right ventricular infarction: recent data and future perspectives]. PMID- 3051307 TI - [Neuro-endocrine disorders in cardiac insufficiency: therapeutic consequences]. PMID- 3051308 TI - [History of the transtympanic drain]. PMID- 3051309 TI - [Primary hyperparathyroidism. History]. AB - The term primary hyperparathyroidism currently refers to the clinical and biological manifestations resulting from the hypersecretion of parathyroid hormone by one or several parathyroid adenomas. This entity is a recent one since it goes back to 1925. The clinical picture resulting from this anomaly, were first described as Recklinghausen's fibrous osteitis, which was not justified since Recklinghausen had not established the relationship between the clinical manifestations and the adenoma discovered by Mandl, then under the name of parathyroid osteosis. This term was justified at a time when the disease presented only bony manifestations with biological evidence. Primary hyperthyroidism is the current appellation, demonstrating that the disease is now a biological disease, often discovered systematically, with therapeutic progress, progress in biological identification and possibility of medical forms without mandatory surgical outcome. Bibliographic references accompany the various stages of this historical reminder. PMID- 3051310 TI - [Primary hyperparathyroidism. Recent physiopathological data]. AB - Recent advances in the physiology of the parathyroid hormone permit a better demonstration of the mechanisms of inappropriate hypersecretion which characterizes hyperparathyroidism, even through the etiology of primary forms still remains unknown. There have been also some progress achieved understanding and evaluating the impact od hypersecretion on target organs, especially bone and kidney. Some studies give an indication of the therapeutic modalities which would enable to reduce the excess of secretion, to counteract the effect of the hormone on its receptor sites or finally limit its consequences on osteopenia. PMID- 3051311 TI - [Surgery of primary hyperparathyroidism in 1988. Strategies during primary operations, in case of failure and in case of cancer]. AB - In primary hyperparathyroidism, an operation is indicated when the calcemia exceeds 115 mg/l and phosphoremia is low in several successive instances, regardless of the symptoms, even if PTH levels and cervical ultrasonography are normal. In case of calcemia under 110 mg/l, the diagnosis must be confirmed by titration of the nephrogenic cyclic AMP and symptomatic patients must be operated upon as well as asymptomatic patients with a life expectancy exceeding 10 years. In case of acute hypercalcemia, the procedure must be performed as a semi emergency without waiting for a definite diagnosis, since the course may rapidly be fatal in spite of all medical treatments. Ultrasonography mostly presents the advantage of detecting intrathyroid parathyroids glands. The experience of the surgeon is essential for the mandatory locating of 4 glands, and the choice of the surgical strategy. In front of a secondary indication for failure or recurrence, one must take into consideration what was seen and done during the first procedure, the calcemia levels and clinical, radiological or biological consequences. Finally, in case of cancer (2 p. cent), the best prognosis rests in wide excision of the thyroid compartment and nodes areas, since medical treatments and radiation therapy are ineffective. PMID- 3051312 TI - [Value of thallium-technetium scintigraphy in the preoperative localization of parathyroid adenoma. Apropos of 30 cases]. PMID- 3051313 TI - [Joint chondrocalcinosis and primary hyperparathyroidism]. PMID- 3051314 TI - [Familial hyperparathyroidism. Excluding polyendocrine adenomatosis]. PMID- 3051315 TI - [Role of interleukin 1 in lymphocyte migration during rheumatoid synovitis]. PMID- 3051316 TI - [The new clinical picture of primary hyperparathyroidism. Diagnostic circumstances and current symptomatic characteristics. Results of a multicenter study]. AB - Primary hyperparathyroidism (PHP) is particularly interesting at this time because of the modifications of its traditional symptoms, which have renewed the conditions of its diagnosis. This is the result of a better knowledge and consequently an increased frequency of the simple forms, mild or clearly atypical, usually expressing the initial stages of the disease which are now better detected. The relative part of the classical manifestations of the disease is therefore reduced as well. In order to verify this fact, the authors have initiated a retrospective study of 535 recent cases of PHP, over 12 years, comparing them with 322 older cases, examined and operated upon between 1954 and 1976 by P.L. Chigot. Analysis of the differences that were noted, was the subject of a statistical evaluation. The first result of this investigation is that PHP remains a disease affecting predominantly women, especially between the ages of 40 and 60 years, and beyond that age to a lesser degree. In comparing the data obtained from analyzing the circumstances of discovery of the disease and its symptoms, the most striking modification consisted in a real drop in the frequency of bony lesions. These modifications are much more rare, only exceptionally presenting their classical characteristic X-ray appearance and they only represent a factor of contingency in the clinical picture of PHP. This is probably the result of a much earlier discovery of the disease. The incidence of renal insufficiency is also remarkably low, probably for the same reason. In return, asthenia and urinary lithiasis are now the major symptoms of PHP.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3051317 TI - [Cervical spondylolisthesis of congenital origin. Apropos of a case, review of the literature]. PMID- 3051318 TI - [Palmar-plantar pustulosis and arthritis of the hands. Apparent and actual analogies to psoriatic arthropathy. Apropos of a new case]. PMID- 3051319 TI - [Acute renal insufficiency and non-steroidal anti-inflammatory drugs]. AB - Non-steroid anti-inflammatory drugs cause an increasing number of renal disorders because of their increasing use. In addition to a toxic or immuno-allergic mechanism common to many medications, the side-effects are essentially related to the inhibition of prostaglandins biosynthesis which play a major role as soon as the renal blood flow is compromised. Acute renal failure of hemodynamic origin, interstitial nephropathy with or without nephrotic syndrome are the most characteristic manifestations of the renal toxicity of non-steroid anti inflammatory drugs. PMID- 3051320 TI - [Genioplasty, indications and therapeutic methods in dento-maxillary malformations]. AB - Genioplasty constitutes an adjunctive surgical procedure used in adult plastic surgery of the facial profile. It is used after dental malocclusion has been corrected and facial morphological features have been reassessed (soft tissue changes consecutive to maxillary osteotomy should be previously analyzed by cephalometry). Genioplasty permits achieving a modification of the chin point in the three planes. The authors describe several types of genioplastic operations, depending on the aim. PMID- 3051321 TI - [The value of comparison of echography and sialo-tomography in the preoperative evaluation of parotid tumors]. AB - Studying information provided by sialo-tomography and parotid ultrasonography in the pre-operative investigation of parotid tumors, the authors compare results provided by the two types of investigation. Whilst ultrasonography, a non invasive technique, provides more detail concerning the topography and appearance of the tumor than sialography, it cannot yet be used as a substitute for this radiological investigation. The two would appear to be complementary, leading the last word to histopathology. PMID- 3051322 TI - [Extra-osseous mandibular ameloblastoma]. AB - Gingival ameloblastoma is a rare odontogenic tumor which shares the same histology as endo-osseous ameloblastoma but from which it differs in terms of its course by the absence of "local malignancy". There would appear to be two probable origins concerning this lesion: Serre debris and the epithelium of the gingival mucosa. However, the etiopathogenesis remains controversial, raising problems of pathological distinction from gingival basal cell carcinoma. PMID- 3051323 TI - Epidermal growth factor content of submandibular glands is increased in rats with experimentally induced gastric lesions. AB - Occurrence and growth-promoting effect suggest that epidermal growth factor (EGF) is involved in the maintenance of the gastrointestinal mucosa. The aim of this study was to ascertain whether rats with gastric lesions, induced by cold-water stress, ethanol, or indomethacin, have altered EGF levels in their gastrointestinal tract compared with controls. For this purpose we established a solid-phase enzyme immunoassay and measured the amount of immunoreactive EGF (IR EGF) in extracts of the submandibular gland, stomach, duodenum, jejunum, ileum, and colon. In the submandibular glands of control rats the IR-EGF content was 100 400 times higher than in the gastrointestinal tract. In animals with experimentally induced ulcer the IR-EGF content was increased in the submandibular glands (1480 versus 423 ng/g), duodenum (11.4 versus 5.8 ng/g), and colon (4.1 versus 1 ng/g), whereas no change was observed in the stomach and jejunum. When the formation of lesions was prevented by omeprazole (stress) or prostaglandin E2 (ethanol and indomethacin), the IR-EGF increase in submandibular glands and duodenum was abolished. This indicates that mechanisms regulating the synthesis and secretion of submandibular EGF might be of importance in the development of gastric and duodenal ulcers. PMID- 3051325 TI - ["Maximal emission values" for air pollutants--definition and viewpoints in prevention]. PMID- 3051324 TI - Effect of pancreatic glucagon and its 1-21 fragment on gastric emptying in man. AB - The effect of pancreatic glucagon (G) and its 1-21 fragment (G 1-21) on gastric emptying was studied in nine healthy volunteers. Gastric emptying of a 200-ml nutrient liquid meal was assessed during continuous infusion of physiologic saline, G, or G 1-21 in equimolar concentrations. The subjects were studied three times on separate days in randomized order. Gastric emptying was measured with a gamma camera technique. The emptying was diphasic in all studies, showing an initial plateau lasting 10-15 min followed by an exponential decline. During saline infusion the time necessary for 50% of the meal to leave the stomach was 32 +/- 4 min, compared with 30 +/- 4 min and 35 +/- 3 min during infusion of G and G 1-21, respectively. It is concluded that glucagon and its 1-21 fragment in physiologic concentrations do not seem to participate in the control of gastric emptying of a nutrient liquid meal. PMID- 3051326 TI - [Guidelines for inside air]. PMID- 3051327 TI - [Responsibilities and principle concerns of environmental medicine]. PMID- 3051328 TI - [Limit values in drinking water]. PMID- 3051329 TI - [Considerations for decreasing the risk in environmental microbiology]. PMID- 3051330 TI - [The significance of inhaled pollutants for bronchial asthma and chronic bronchitis]. PMID- 3051331 TI - International experience with tenoxicam: a review. AB - For new drugs with relatively few side-effects, a large side-effect database needs to be gathered before a realistic appraisal of a drug's side-effects profile can be made. The database can be collected in different ways. A United Kingdom database of 4,125 patients receiving tenoxicam and 1,132 patients receiving comparator drugs requires all events, even if only remotely related to therapy, to be reported. An international database, currently numbering 5,537 patients receiving tenoxicam and 2,948 patients receiving comparator non steroidal anti-inflammatory drugs (NSAIDs), is less stringent; only events felt by the physician to be directly related to the drug are reported. Both these databases are open. A separate randomised, double-blind, UK study compared 657 patients on tenoxicam 20 mg/day and 663 patients on piroxicam 20 mg/day. All three methods of data collection show a similar trend which is that tenoxicam causes fewer serious gastrointestinal side-effects than piroxicam, although where homogeneity of population allows the application of statistics, this does not achieve statistical significance. In all other respects the side-effects profile of tenoxicam was virtually identical to that seen with piroxicam and similar to that seen with other NSAIDs. PMID- 3051332 TI - Local glycosaminoglycan polysulphate injection therapy in osteoarthritis of the hand. A placebo-controlled clinical study. AB - In this placebo-controlled double-blind trial in 30 patients suffering from disabling osteoarthritis of the hand, the long-term effect of periarticular injections of glycosaminoglycan polysulphate (GAGPS) on symptoms, hand function and life quality was investigated. The overall effect of the half-year follow-up was clinically good in 46% of the GAGPS-treated cases, compared with 14% of the controls. Significant differences in grip- and pinch strength were also found between the treatment groups. Significant differences were still seen at the one year follow-up, as 13 patients (87%) in the GAGPS group reported improvement in their most restricted activity, compared with 6 (43%) of the controls. Nine periarticular injections of GAGPS over a period of 13 weeks gave a clinically good and long-lasting effect, with improved life quality for most of the patients. PMID- 3051333 TI - Immunocytology of synovial fluid cells. A prognostic indicator in inflammatory arthritis? PMID- 3051334 TI - Health hazards of tertiary amine catalysts. AB - Tertiary amine catalysts are widely employed in foundry and polyurethane foam manufacture operations. These highly reactive amines have been associated with graphic disturbances in vision and systemic health effects. Prominent among the reported effects on vision are mydriasis (dilated pupils), cycloplegia (loss of accommodation), and corneal edema, which may result in hazy (looking through smoke) or blurry (out of focus) vision and halo perception. Systemic symptoms, possibly due to a release of endogenous histamine, are consistent with pharmacologic actions of amines and have also been described. These symptoms, as well as the disturbances in vision, are transient. Nevertheless, employees who work with or around machinery, or drive vehicles, may be at an increased risk of accident and injury when experiencing these symptoms. PMID- 3051335 TI - [Sleep-apnea syndrome. Elucidation, therapy and course]. AB - Of 22 patients investigated for sleep disorders, habitual snoring and/or daytime hypersomnolence, 12(10 men) had obstructive sleep apnea syndrome (OSAS). 3 OSAS were mild, 5 moderate and 4 severe. The leading symptoms were daytime hypersomnolence and habitual snoring. As risk factors we found retro-micrognathia in 2 patients, macroglossia secondary to acromegaly in 1, alcohol abuse in 7 and obesity in 6. Conservative measures improved the disorder subjectively in 6 patients. One patient had a relapse 6 months after uvulopalatopharyngoplasty. 4 patients were successfully treated by nasal CPAP. Other diagnoses were idiopathic alveolar hypoventilation (2), Cheyne-Stokes breathing secondary to low cardiac output (1), monosymptomatic narcolepsy (2), sleep disturbances secondary to depression (2), chronic benzodiazepine abuse (1) and chronic bronchitis without nocturnal hypoxemia (1). History, clinical observation and oxymetry make diagnosis possible in most cases of OSAS severe enough to require treatment. Polysomnography is time-consuming and should be reserved for selected cases. PMID- 3051336 TI - [Switzerland changes from U-40 to U-100 insulin]. AB - At present over 95% of the insulin used in Switzerland is sold in a concentration of 40 units per ml (U-40 insulin). Only recently a small but increasing amount of more concentrated insulin has been introduced through the use of "pen" injectors which are all designed for U-100 (= 100 units per ml) insulin. Experience in other countries has shown that when different insulin concentrations are available on the market the risk of dosage errors with potentially severe consequences is high. The Swiss Diabetes Association has therefore taken the initiative in standardizing the concentrations available in Switzerland. A change from U-40 and U-100 to exclusively U-100 insulin is planned for the period from October 1988 to March 1989. A survey among 82 Swiss doctors and 243 IDDM patients has shown that about 60% of doctors and patients are in favour of the change while the remaining 40% are either neutral or opposed to it. Advantages, disadvantages and logistic problems of the change to exclusive use of U-100 insulin, and the possibility of pharmacological differences between insulins of different concentrations, are discussed. It is stressed that the change to U-100 must be medically supervised in every patient. An intensive information campaign providing detailed advice for patients, doctors and pharmacists is essential for the prevention of accidents. PMID- 3051337 TI - [Does the mind drift where it wants? Requirements for medical discoveries]. AB - In medical discoveries, two elements may be distinguished: (1) the state of science and the available methods and tools of research at a given moment, and (2) the personal and original intellectual achievement. The relative importance of these two factors is discussed in the light of historical examples. It is apparently rare that a discovery results more or less automatically from a given state of science and research (as in the case of insulin). Nearly always the personality of the research worker turns out to be the determinant factor. Essential traits of this personality are an independent mind capable of liberating itself from dogmatic tenets universally accepted by the scientific community; the capacity and courage to look at things from a new angle; powers of combination, intuition and imagination; feu sacre and perseverance--in short, intellectual as well as moral qualities. The personality of a great scientific discoverer, furthermore, integrates the spirit of his epoch into his individuality. About half the research workers considered here were aged below 35 at the time of their discoveries. The author therefore hopes that our universities will, in the future, provide opportunities for young research workers as well as for somewhat older individualists. PMID- 3051339 TI - [Drug therapy in peripheral arterial occlusive disease]. AB - Drugs used for improvement or stabilization of peripheral circulation include antiaggregants or anticoagulants for secondary prevention of arteriosclerosis, vasoactive substances and fibrinolytic agents. -Two prospective trials document that aspirin or the combination of aspirin and dipyridamole reduce progression of arterial occlusive disease significantly in comparison to placebo. Aspirin is best suited for secondary prevention of recurrent stenoses or occlusions after carotid or femoral endarterectomy, whereas anticoagulants are preferred in patients with embolism and after peripheral implantation of venous bypasses. Significant improvement of walking distance is achieved by several compounds influencing blood rheology. The effect does not exceed that obtained by physical training. If reconstructive arterial surgery or percutaneous transluminal angioplasty are not possible, some patients with rest pain or gangrene may be successfully treated by intraarterial administration of prostaglandin E1. PMID- 3051338 TI - [Malaria in Switzerland: 1982-1986]. AB - There has been no decrease in the number of cases of malaria brought into Switzerland. 841 cases were reported to the Federal Office of Public Health between 1982 and 1986, a figure probably below the actual number of cases. Reports were received from all cantons and the majority involve Swiss travellers. 54% of the cases were due to P. falciparum and 90% of these could be traced to a stay in tropical or subtropical Africa, particularly Kenya, where it was possible to determine the attack rate and to compare it with the risk in Thailand. An enquiry into prophylaxis disclosed how slowly the latest recommendations on preventive measures against malaria are being adopted, and the importance of keeping a close watch on regions where resistance or multiresistance has developed. Systematic reporting is essential to control and also permits the recommendation of appropriate preventive measures. PMID- 3051341 TI - [Clinical spectrum of a common and insidious pathogen: Streptococcus milleri]. AB - We studied the clinical significance of S. milleri isolated in our hospital in 68 patients during a 18-month period. In 51 patients (median age: 43 years, no underlying diseases in 29 patients), the isolates were associated with significant infections. They were beta-hemolytic in 32 cases and non-hemolytic in 19. The primary infection sites were the head and neck area (21 cases), the lungs (5 cases of pneumonia), the gastrointestinal tract (12 cases), the urogenital tract (3 cases), the soft tissues (6 cases), and the heart (2 endocarditis). Two septicemias were of unknown origin. Head and neck infections and pneumonia were most often associated with beta-hemolytic strains, and bacteremia, gastrointestinal and urogenital tract infections with alpha-hemolytic strains. S. milleri was found in pure culture in 24 cases. Polymicrobial associated flora (27 cases) was more frequent in the abdominal infections (87%) than in supra diaphragmatic infections (42%). Severe complications were observed in 12 head and neck infections (57%) (cerebral abscesses 3, lethal mediastinitis 2, osteitis 1, meningitis 1, other suppurative lesions 5). When abscesses were present (27 cases), surgery was required in all cases. Despite the high frequency and severity of local complications, the clinical outcome was usually favorable. However, deaths directly related to S. milleri infections occurred in 2 cases of mediastinitis complicating the course of apparently harmless primary infections. Owing to the possible occurrence of life-threatening complications, S. milleri infections require early identification, treatment and surgery when indicated. PMID- 3051340 TI - [MTBE litholysis and extracorporeal shock wave lithotripsy of gallstones]. AB - Two new therapies for cholesterol gallstones, MTBE litholysis and extracorporeal shock-wave lithotripsy, have been developed and clinically tested in the last five years. In principle MTBE litholysis is applicable in 50-80% of gallstone patients and shock-wave lithotripsy in 25-30%. In MTBE litholysis a pigtail catheter is placed in the gallbladder by the percutaneous-transhepatic route using ultrasound-guidance. The gallstones can than be dissolved with the cholesterol contact solvent MTBE (methyl tert-butyl ether). With appropriate selection criteria the successful dissolution rate is 80-90%. Over 99% of radiolucent gallstones (1-3 stones per gallbladder) can be fragmented by extracorporeal shockwaves. Bile acids must be administered as adjuvant litholytic therapy, and thus the remaining fragments can be dissolved in 91% of cases within 2-18 months. In view of the limited experience with these procedures at the present time they are mainly indicated for symptomatic surgical high-risk patients and for those who strongly wish to avoid surgery. With careful evaluation of study data a broader indication policy may be possible in the future. The new methods and the essential study data are presented. PMID- 3051342 TI - [Results of resection treatment of locally limited bronchial carcinoma (Stages I and II)]. AB - Of 238 patients operated upon for bronchogenic carcinoma between 1977 and 1985 the tumor was locally limited in 108, of whom, according to the new international staging system, 66 patients were in stage I (T1N0 + T2N0) and 42 in stage II (T1N1 + T2N1). 89 were male and 19 (18%) female. The mean age was 61.6 years in stage I und 63 years in stage II patients. 26 patients (24%) were aged over 70. All patients underwent potentially curative excision of the tumor. This was accomplished by partial lung resection in 97 patients, of whom 84 underwent lobectomy with 2 postoperative deaths (2.4%). Operative mortality amounted to 5.5% in the whole group (6/108), with 3% (2/66) in stage I and 9.5% (4/42) in stage II patients. The absolute survival rate at 5 years was 50% for all 108 patients, 59% for the 66 patients in stage I and 39% for the 42 patients in stage II. It was 70% for 15 patients with bronchioloalveolar carcinoma, 54% for 20 patients with adenomatous cancer and 49% for 56 patients with squamous carcinoma. At present 62 of the 108 patients are still alive (57.4%), i.e. 45 out of 66 in stage I (68%) and 17 out of 42 in stage II (40.5%). This retrospective study therefore confirms former reports that long term results are encouraging in the locally limited stages of pulmonary cancer. In addition, operative mortality is low because almost all these tumors can be removed by partial lung resection. In stage I lobectomy is the method of choice. PMID- 3051343 TI - [Strategically important abutments]. PMID- 3051345 TI - [Dental cement, that forgotten substance. A review of the literature]. PMID- 3051346 TI - The 57th variety. PMID- 3051347 TI - How killer cells kill. AB - Killer lymphocytes, the commandos of the immune system, attack tumor cells and cells infected by viruses. They kill by secreting protein molecules that link to form pores in target cells; the cells promptly leak to death. Study of the process may make it possible to improve the killers' efficiency in fighting cancer and such viral infections as AIDS. PMID- 3051344 TI - [Oral hygiene in older patients. Psychological and practical aspects of oral hygiene in gerodontology]. PMID- 3051348 TI - Art, illusion and the visual system. AB - The verve of op art, the serenity of a pointillist painting and the 3-D puzzlement of an Escher print derive from the interplay of the art with the anatomy of the visual system. Color, shape and movement are each processed separately by different structures in the eye and brain and then are combined to produce the experience we call perception. PMID- 3051349 TI - The amateur scientist. PMID- 3051350 TI - Dial-a-doc. PMID- 3051351 TI - Catalytic antibodies. PMID- 3051353 TI - Heartening news. PMID- 3051352 TI - Fatness and fertility. PMID- 3051354 TI - Tumor necrosis factor. AB - A century ago it was noted that a bacterial infection sometimes causes the regression of cancer. In 1975 the author found an explanation: the infection stimulates the secretion of tumor necrosis factor (TNF), which has anticancer activity. Now TNF, an important regulator of inflammation and immunity, is in clinical trials as an anticancer drug. PMID- 3051355 TI - The neurobiology of feeding in leeches. AB - How does a simple nervous system control a behavior? In the bloodsucking medicinal leech a single neurotransmitter, serotonin, has been found to orchestrate the animal's search for a target, the movements of its jaws, the filling of its crop and even the distension of its body that eventually tells the leech enough is enough. PMID- 3051356 TI - [Sonography in cystoid cervical space occupying lesions]. AB - A retrospective analysis of fluid-filled cervical space-occupying lesions is performed. The high echogenicity of some cervical cysts is due to the difference of the acoustic impedance of water and agglomerated cholesterol crystals. A sonomorphological differential diagnosis of cystoid cervical tumors and fluid accumulations is performed. Uncommon criteria of cysts and new techniques for the differentiation of cystoid cervical lesions are discussed. The clinical role of sonography in the diagnosis of cervical cysts and cystoid lesions is evaluated on the basis of 180 histologically proved cases. PMID- 3051358 TI - [Changes in renal echogenicity in pediatric obstructive uropathies]. AB - Renal cortical echogenicity of 40 children with obstructive uropathies was analysed in relation to the grade of obstruction and thinning of the renal cortex. In 35 percent of the children an increase in echogenicity was noted. The increase in cortical echogenicity paralleled the grade of dilatation of the intrarenal collecting system and the grade of thinning of the renal cortex. Evaluation of the cortical echogenicity should be integrated into the primary diagnostic setup as well as into the follow-up of children suffering from obstructive uropathies. PMID- 3051357 TI - [Sonography of adrenal gland diseases in the newborn infant]. AB - Diseases and morphological changes of the adrenal gland play an important role especially in the newborn, because malformations, congenital errors of metabolism, connatal tumours and birth injuries become symptomatic in this period of life. In this paper, the possibilities of neonatal adrenal ultrasonography for diagnosis and in the aspect of differential diagnosis are discussed. The characteristic sonographic appearance is the thin echogenic core surrounded by a thick transonic zone. Adrenal haemorrhage as well as the neuroblastoma disturb these structures and show their own characteristic pattern. In congenital adrenal hyperplasia the organ is enlarged in a fold-like manner. In agenesis or displacement of the kidney the adrenal gland presents as an elongated mass in the renal fossa and on the psoas muscle. PMID- 3051359 TI - [Ultrasound controlled kidney cyst puncture: significance for diagnosis and therapy]. AB - Renal cyst puncture under ultrasonic guidance has proved an excellent and safe method in the differential diagnosis of cystic renal masses. The use of ultrasound in the follow-up of patients with renal cyst puncture is mandatory for evaluation of complications and results. The therapeutic success of renal cyst puncture is poor and estimated at about 25%. New percutaneous techniques with resection of the cystic wall lead us to expect much better results than with open surgical management. PMID- 3051360 TI - [The development of standards for ophthalmologic ultrasound diagnosis]. AB - The performance data of ultrasonic apparatus and transducer probes are decisive in view of the information content of the echograms. Clinically applicable measurement methods, therefore, were developed already in the early sixties for ophthalmic ultrasonography. The sensitivity of each probe apparatus combination was measured first by means of a variable absorption path, the working frequency by counting the oscillations per microsecond in the rf echo signal of a plane reflector. Depth and lateral resolution were determined using monofil filaments mounted to a micrometer gauge. These measurements resulted in marked improvement of safety of diagnosis. In view of the IEC document 854, we developed a test reflector. It reflects a standard echo which can be measured within the control range of most diagnostic machines without need of additional dB-calibrated attenuators. Now, each echo amplitude can be referred to this standard echo (whose difference to the ideal reflector echo is-17 dB). An electronic device (echo simulator) permits additional checking of distinct equipment parts (especially of the dB control, time scale and amplifier dynamics). The entirely empirical use of ultrasonic diagnostic apparatus without measurement of the diagnostically relevant performance data and without readjustment and documentation of comparable examination conditions is, in our opinion, an outdated approach. Repeated measurements at regular intervals and especially after repairs or change of transducer probes are mandatory. PMID- 3051361 TI - [Angiodynography: technical principles and clinical uses]. AB - Angiodynography is a newly developed system that enables Doppler sonography to be used to obtain a colour-coded real-time image of the blood vessels in the familiar ultrasound slice image. A new computerised technique allows both the familiar pulse echo signal and the Doppler shift of the signal resulting from movement in the body to be displayed for each point on the ultrasound slice plane. Stationary tissue structures are shown as the familiar grey scale image, while movement such as of the blood is shown in colour. 453 patients were examined. Normal and pathological flow measurements were obtained in the carotid artery, the jugular vein, in renal transplants, in the thyroid, the testis and in the urethra. Pathological flow velocities were visualised in stenosis, tumours and diffuse alterations of parenchyma. PMID- 3051362 TI - [Synthetic ultrasound blocks and contact gels in sonography: hygienically satisfactory?]. AB - Different sponge-like plastic blocks and several samples of two coupling gels used during small-parts real-time sonography were investigated microbiologically. Whereas the coupling gels showed no contamination a great deal of different bacteria could be found on the plastic blocks. These findings lead to a restricted use of those devices especially when investigating immunocompromised patients. Sufficient procedures of decontamination do not exist. Because of the relative high purchase cost the one-way use of plastic blocks is uneconomical. PMID- 3051363 TI - [What does ultrasound study contribute in acute appendicitis (using dosed compression)?]. AB - The value of ultrasound in diagnosing acute appendicitis has been examined in a prospective study of 103 patients. Only those patients with an uncertain diagnosis of acute appendicitis were included in the study. We used a 5 or 7.5 MHz linear array transducer applying graded compression. An acutely inflammatory appendix appeared as a tubular structure with a diameter of 3 to 12 mm. The sensitivity of the examination in our material was 82%, the specificity 97%. The main reason for the relatively low sensitivity were cases with a retrocaecal position of the appendix. Ultrasound can be recommended as a highly specific but less sensitive imaging method for the diagnosis of acute appendicitis in clinically equivocal cases. PMID- 3051364 TI - [Phlegmonous inflammation of the large intestine: a sonographic diagnosis?]. AB - Diseases of the gastrointestinal tract accompanied by a thickened bowel wall have become more accessible to ultrasonic detection during the last years. In most of the cases, however, the sonographic pattern has no characteristic feature for any disease. In this paper we present the rare case of a phlegmon of the colon and discuss its differential diagnosis. PMID- 3051366 TI - [Ultrasonography in the primary diagnosis and follow up of ovarian tumors]. AB - Preoperative sonographic data in 317 patients who underwent surgery at the University Women's Clinic Heidelberg for an ovarian tumour during a 6 years' observation period, were compared with the results of clinical examination. The prognostic value of both techniques was examined, by relating the findings (as to tumour size, demarcation from the uterus) to the postoperative histological or cytological diagnosis. In the 214 cases of benign and 103 of malignant ovarian tumours a correct diagnosis was provided by ultrasound in 88% and 74% respectively. Thus significantly more often than by clinical examination alone (p less than 0.001 for benign and p less than 0.01 for malignant tumours). The sonographic findings in 49 patients during follow-up after surgery for ovarian carcinoma and polychemotherapy were checked by a second look laparotomy. The detection rate of tumour progression, recurrence, residual tumour or remission was of 98%. PMID- 3051367 TI - [Sonographic volumetry of pleural effusions]. AB - Chest sonography is more sensitive than conventional x-ray examination in diagnosis of pleural effusion. We developed a method for an easy quantification of pleural effusion in the sitting position by using simple sonographic parameters. The validity of the method was tested in 17 consecutive patients by pleural evacuation after thoracocentesis. PMID- 3051369 TI - [Circulation studies and transcranial Doppler sonography in orthostatic regulation disorders]. AB - The effect of postural blood pressure changes on the blood flow velocity in the middle cerebral artery (MCA) was studied with transcranial Doppler sonography in 45 patients with intact circulatory reflexes and in 30 cases with postural hypotension. In an upright position on the tilt-table the patients with intact cardiovascular reflexes showed a small decrease of the MCA flow velocity of 3% (1.5 cm/sec), whereas there was found a marked reduction in the cases with postural hypotension. In the group with an asympathicotonic response (n = 14) there was an average decrease of 24% (13 cm/sec) and a 28% drop of mean arterial blood pressure. The lower percentage of reduction of the cerebral blood flow velocity can be explained as an effect of cerebral autoregulation. In the group with a sympathicotonic response (n = 16) a surplus reduction of the flow velocity of 17% (7 cm/sec) was seen, compared to a 7% decrease in blood pressure. This corresponds with the results of experimental animal studies which showed an impaired cerebral autoregulation during increased sympathico-adrenal discharge. PMID- 3051365 TI - [Sonography as a screening examination method of the venous system in pregnancy. Comparative studies of the arm and leg vein system]. AB - Comparative studies were conducted during the third trimenon of pregnancy between the 36th and 40th weeks of gestation and on the seventh day after delivery, at the venous system of the legs (vena iliaca, v. femoralis and v. poplitea) and at the venous system of the arms (v. brachialis). A marked reduction of the venous diameters was seen on the legs, whereas no difference was found in the venous system of the arms between both times of measurement. This points to the haemodynamic effect of the pregnant uterus, providing an obstacle to the flow of blood from the venous system of the legs. After delivery there is a relatively higher extensibility of the veins of the leg compared with the measurements in the third trimenon. This may be due on the one hand to the long-lasting overextension during pregnancy, and on the other hand also to the hormonal and enzymochemical changes that take place even during the puerperium. Doppler sonography of the v. femoralis showed clearly that the foetus obstructs blood flow from the venous system of the legs. Sonography seems to be able to visualise direct the changed morphology of the veins during pregnancy and to indirectly yield information on the extensibility of the venous system eg during standing, thus demonstrating in what way the venous system functions in a particular condition. Hence, sonography of the venous system of the legs during pregnancy should be used as a screening method wherever it is essential to discover extreme venous dilatations well in time and to initiate compression treatment. PMID- 3051368 TI - [Flow measurements in extracranial carotid arteries by means of duplex sonography. Results in normal subjects]. AB - Carotid arteries were examined by Duplex sonography in 116 healthy volunteers of between 16 and 78 years of age. We measured the internal carotid artery and common carotid artery peak systolic velocities and their ratio, the common carotid artery time-averaged velocity, the maximal diameter of the common carotid artery, and the common carotid artery volume flow (ml/min). Flow velocities and volume flow showed an age-dependent decrease. PMID- 3051370 TI - [Duplex sonography in carotid-cavernous sinus fistula]. AB - The absence of valves in the ophthalmic veins leads to a reversal of flow and marked vascular dilatation if there is a carotid-cavernous sinus fistula. Based on a case report, the possibility of accurate duplex sonographic evaluation of these findings is shown. Duplex sonography should be employed as a simple, non invasive and inexpensive diagnostic method even if the likelihood of a carotid cavernosus sinus fistula is remote. PMID- 3051371 TI - [Doppler sonographic characterization of vessels of the posterior cranial fossa using a suboccipital paramedian approach]. AB - By means of transcranial Doppler sonography we examined the posterior cranial fossa from the suboccipital paramedian region. This enabled us to identify the basilar artery in its entire course, as well as the posterior inferior cerebellar artery. The advantages of this approach are, in particular, the possibilities of examining postoperatively by Doppler sonography after medial surgical approach, as well as the future utilisation the technique within the overall framework of stereometric transcranial Doppler sonography. PMID- 3051372 TI - [Doppler sonography findings in arteriitis temporalis]. AB - The results of 145 patients who had been examined by Doppler sonography for suspected giant cell arteritis were analysed. Alternating flow direction, reversed flow direction or "no-flow" in the supratrochlear and/or supraorbital arteries were considered as pathological findings as well as absence of signals from the trunk or branches of one or both superficial temporal arteries. Pathological findings at the periorbital arteries were obtained in 25.9% (n = 7) of the cases with histological proven giant cell arteritis. In a further 29.6% (n = 8) the superficial temporal artery findings were abnormal. Only 13.2% (n = 5) of the patients with normal histological findings and 5.7% (n = 2) of those with degenerative lesions of the temporal arteries had pathological Doppler sonographic findings. Two thirds of the patients with visual impairment showed abnormal findings in the periorbital arteries. PMID- 3051373 TI - [Sonographic detection of fasciculations]. AB - Fasciculations are an important neurological sign of spinal muscular atrophies. 26 patients able to walk suffering from adult onset spinal muscular atrophies were examined by real time sonography. On an average 18 limb and extremity muscles on both sides were examined. In 21 cases fasciculations became sonographically apparent as muscular twitchings lasting for 0.2 to 0.5 seconds. Detection of fasciculations in profound muscular layers and in obese patients was easier by sonography than by clinical examination. PMID- 3051374 TI - [Transabdominal lumbal ultrasonic tomometry--sonographic measurement of the lumbar spine]. AB - Transabdominal sonography was carried out on 102 lumbar segments that were myelographically without abnormalities of the dural sack. A sector probe with a frequency of 3.5 mHz was used as transducer. Diameters of lumbar discs and lumbar canal were measured in a transversal plane. Echogenicity of the epidural fat in the lumbar canal allows to depict both the limitation of the dorsal outline of the lumbar discs and the dorsolateral vertebral arch. The following mean values were calculated for the spinal canal: Sagittal diameter-L5/S1:18.5 mm, L4/5:20.4 mm, L3/4:19.9 mm Transversal diameter-L5/S1:20.9 mm, L4/5: 21.4 mm, L3/4:21.2 mm Sonographic measurement of the spinal canal is possible. PMID- 3051375 TI - Spontaneous subclavian vein thrombosis. AB - A young man presented with swelling of his right arm and engorgement of the superficial veins over the shoulder. Bilateral upper limb venography was performed and confirmed the presence of thrombus in the right subclavian vein. Deep venous thrombosis (DVT) of the upper extremity is an unusual thrombotic event with a reported incidence of 1-2% of all cases of venous occlusion. Review of the literature reveals a number of diverse aetiologies, and no agreement regarding the best terminology for the subgroups of this disorder. PMID- 3051376 TI - Great doctors and medical worthies. AB - After Harvey's visits to Scotland with Charles I the formation of a united Caroline University in Aberdeen was thwarted by the Civil War. In Oxford Harvey instituted a group of medical scientists, forerunners of the Royal Society, who almost explained the physiology of respiration. Harvey had several things in common with Dr Samuel Johnson. Johnson's medical knowledge and contacts are emphasised, examples of 17th and 18th century health regimens are given and Johnson's friendship with Scottish medical men and some others connected with the Royal College of Physicians and the Harveian Society of Edingburgh are described. PMID- 3051377 TI - Induction of B cell unresponsiveness to noninherited maternal HLA antigens during fetal life. AB - Patients who have received many transfusions become highly sensitized and develop antibodies against almost all HLA alloantigens, so that finding a cross-match negative kidney donor is difficult. A survey of those patients showed that 50 percent did not form antibodies against the noninherited maternal HLA antigens. Apart from the obvious clinical implications, the data indicate that a human equivalent of murine neonatal or actively acquired tolerance has now been identified. PMID- 3051378 TI - Human IL-3 and GM-CSF act synergistically in stimulating hematopoiesis in primates. AB - Interleukin-3 (IL-3) is a member of a family of growth factors, each of which supports the proliferation and development of hematopoietic precursors in culture. Although the biologic effects of the different hematopoietic growth factors have been well documented in different culture systems, it has only recently become possible to study the activities of these molecules in vivo. In comparison with the later acting hematopoietic growth factors granulocyte macrophage colony-stimulating factor (GM-CSF) and granulocyte colony-stimulating factor, IL-3 elicited a delayed and relatively modest leukocytosis when continuously infused intravenously in primates. The IL-3 infusion, however, greatly potentiated the responsiveness of the animal to subsequent administration of a low dose of GM-CSF. These results suggest that IL-3 expands an early cell population in vivo that subsequently requires the action of a later acting factor such as GM-CSF to complete its development. Optimal stimulation of hematopoiesis may be achieved with combinations of hematopoietic growth factors. PMID- 3051380 TI - Scanning tunneling microscopy and atomic force microscopy: application to biology and technology. AB - The scanning tunneling microscope (STM) and the atomic force microscope (AFM) are scanning probe microscopes capable of resolving surface detail down to the atomic level. The potential of these microscopes for revealing subtle details of structure is illustrated by atomic resolution images including graphite, an organic conductor, an insulating layered compound, and individual adsorbed oxygen atoms on a semiconductor. Application of the STM for imaging biological materials directly has been hampered by the poor electron conductivity of most biological samples. The use of thin conductive metal coatings and replicas has made it possible to image some biological samples, as indicated by recently obtained images of a recA-DNA complex, a phospholipid bilayer, and an enzyme crystal. The potential of the AFM, which does not require a conductive sample, is shown with molecular resolution images of a nonconducting organic monolayer and an amino acid crystal that reveals individual methyl groups on the ends of the amino acids. Applications of these new microscopes to technology are demonstrated with images of an optical disk stamper, a diffraction grating, a thin-film magnetic recording head, and a diamond cutting tool. The STM has even been used to improve the quality of diffraction gratings and magnetic recording heads. PMID- 3051379 TI - tRNAi(met) functions in directing the scanning ribosome to the start site of translation. AB - The mechanism by which the scanning ribosome recognizes the first AUG codon nearest the 5' end of eukaryotic messenger RNA has not been established. To investigate this an anticodon change (3'-UCC-5') was introduced into one of the four methionine initiator (tRNAi(met) genes of Saccharomyces cerevisiae. The ability of the mutant transfer RNA to restore growth properties to his4 initiator codon mutant yeast strains in the absence of histidine was then assayed. Only the complementary codon, AGG, at the his4 initiator region supported His+ growth. The mutant transfer RNA also directed the ribosome to initiate at an AGG placed in the upstream region of the his4 message. Initiation at this upstream AGG precluded initiation at a downstream AGG in accordance with the "scanning" model. Therefore, an anticodon: codon interaction between tRNAi(met) as part of the scanning ribosome and the first AUG must function in directing the ribosome to the eukaryotic initiator region. PMID- 3051381 TI - DNA diagnostics--molecular techniques and automation. AB - Molecular biology has revolutionized the understanding of many aspects of human disease. Ongoing developments in DNA diagnostics--the analysis of disease at the nucleic acid level--will soon provide automated, rapid, and inexpensive analyses for DNA or RNA sequences associated with genetic, malignant, and infectious diseases. DNA diagnostics will also facilitate the identification of disease associated genes at birth, thus creating new opportunities for preventive medicine. PMID- 3051382 TI - Antibodies to Asp-Asp-Glu-Asp can inhibit transport of nuclear proteins into the nucleus. AB - The signal sequence of simian virus 40 (SV40) large T-antigen for translocation into the nucleus is composed of positively charged amino acids Lys-Lys-Lys-Arg Lys. Rabbit antibodies to a synthetic peptide containing the negatively charged amino acid sequence Asp-Asp-Asp-Glu-Asp were obtained. Indirect immunofluorescence of the antigens recognized by the antibody was punctate at the nuclear rim or the nuclear surface, depending on the plane of focus. The antibody blocked transport of nuclear proteins into the nucleus. The antigens recognized by the antibody were predominantly localized to the nuclear pores. PMID- 3051383 TI - Activation of muscarinic potassium currents by ATP gamma S in atrial cells. AB - Intracellular perfusion of atrial myocytes with adenosine 5'-(gamma-thio) triphosphate (ATP gamma S), an ATP analog, elicits a progressive increase of the muscarinic potassium channel current, IK(M), in the absence of agonists. In this respect, ATP gamma S mimics the actions of guanosine triphosphate (GTP) analogs, which produce direct, persistent activation of the guanyl nucleotide-binding (G) protein controlling the K+(M) channel. The effect of ATP gamma S on IK(M), however, differs from that produced by GTP analogs in two aspects: it requires relatively large ATP gamma S concentrations, and it appears after a considerable delay, suggesting a rate-limiting step not present in similar experiments performed with guanosine 5'-(gamma-thio) triphosphate (GTP gamma S). Incubation of atrial homogenates with [35S]ATP gamma S leads to formation of significant amounts of [35S]GTP gamma S, suggesting that activation of IK(M) by ATP gamma S arises indirectly through its conversion into GTP gamma S by cellular enzymes. ATP gamma S is often used to demonstrate the involvement of protein phosphorylation in the control of various cellular processes. The finding that cytosolic application of ATP gamma S can also lead to G-protein activation implies that experiments with ATP gamma S must be interpreted with caution. PMID- 3051384 TI - The 1988 Nobel Prize for Physiology or Medicine. PMID- 3051385 TI - How do enzymes work? AB - The principle of transition-state stabilization asserts that the occurrence of enzymic catalysis is equivalent to saying that an enzyme binds the transition state much more strongly than it binds the ground-state reactants. An outline of the origin and gradual acceptance of this idea is presented, and elementary transition-state theory is reviewed. It is pointed out that a misconception about the theory has led to oversimplification of the accepted expression relating catalysis and binding, and an amended expression is given. Some implications of the transition-state binding principle are then explored. The amended expression suggests that internal molecular dynamics may also play a role in enzymic catalysis. Although such effects probably do not make a major contribution, their magnitude is completely unknown. Two examples of recent advances due to application of the transition-state binding principle are reviewed, one pertaining to the zinc protease mechanism and the other to the generation of catalytic antibodies. PMID- 3051387 TI - Regulation of thrombin generation and functions. PMID- 3051386 TI - Regulation of thrombin generation at cell surfaces. AB - A complex series of reactions are involved in the assembly, function, and regulation of the prothrombinase complex. Since the enzyme is multicomponent in nature and each component is required for catalytic function, modulation of enzymatic activity can be achieved in a variety of ways. In addition, since complex assembly so profoundly affects reaction rates, mechanisms that perturb complex formation either positively or negatively have a profound effect on thrombin generation and its local physiologic effects. All of the cells that support prothrombinase assembly and hence thrombin generation respond to thrombin in a variety of ways. Thrombin selectively binds to thrombomodulin and heparin like molecules expressed on the endothelial cell surface. Thrombin induces the release (and possible synthesis of) prostacyclin, plasminogen activator inhibitor, platelet-derived growth factor, and interleukin-1 and inhibits the release of plasminogen activator from vascular endothelium. Interleukin-1 is a potent mediator of inflammatory phenomena as well as an inducer of tissue factor synthesis in vascular endothelium. With respect to platelets, thrombin selectively binds and stimulates the platelet release reaction and subsequent aggregation. The thrombin-induced release of platelet-derived growth factor from both platelets and vascular endothelium may play a role in inflammation, wound healing, and atherogenesis. Thrombin itself is a potent mitogen of mesenchymal cells, and more recently has been shown to be not only a chemoattractant, but also a mitogen for monocytes. Thrombin also appears to bind selectively to monocytes and in so doing induces release of interleukin-1. Thrombin affects a myriad of cellular responses related to hemostasis, thrombosis, inflammation, would repair, and atherogenesis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3051388 TI - Hyperlipidemia and in vivo hemostatic system activation. AB - Coagulation system activity in vivo is a well-recognized consequence of advanced atherosclerotic disease, the association of coagulation system activity with atherogenesis is much less clear. The data reviewed here argue against ongoing increased coagulation system activity in persons with hyperlipidemia. The possibility remains that episodic activity could easily be missed with available tests, such bursts of activity might well require new agents and strategies for effective control. Failure to find evidence of increased coagulation activity in patients with hyperlipidemia does not refute models of atherogenesis based on thrombosis. Rather, the data reviewed here should cause us to focus attention on the time scale of such events. On the other hand, the findings do allow for a simpler interpretation of data showing evidence of activation of coagulation in patients with concurrent hyperlipidemia. PMID- 3051389 TI - Hypercoagulability and factor VII in hypertriglyceridemia. PMID- 3051390 TI - Vitamin K-dependent proteins bind to very low-density lipoproteins. AB - It has been demonstrated that the surface of large VLDL Sf 100-400 can bind both prothrombin and Factor X(Xa) and that on VLDL Factor Xa can convert prothrombin to thrombin, which degrades apo B and apo E. It has been reported also that the VLDL kinetically supports the conversion of prothrombin to thrombin. The binding of vitamin K-dependent proteins to phospholipid is partially Ca2+-dependent and probably involves their Gla residues. The complex of VLDL, prothrombin, Factor Xa, and Ca2+ lacks only Factor Va, a lipid associating, non-Gla residue containing 330 kd protein, to complete the "prothrombinase complex." Factor V (Va) is found at very low concentrations in the circulation, but is localized on platelets, monocytes, and the endothelium. VLDL can bind both to monocytes and to the endothelium, for example, through both receptor and non-receptor pathways. When carrying this complement of the prothrombinase complex, this subpopulation of VLDL, in the presence of Factor Va on cell surfaces, could conceivably upset the local balance of pro- and anticoagulant activities. Thus, directly or indirectly the increased triglyceride levels, reflected in increased VLDL in patients, may alter this balance, and thereby produce a "hypercoagulable state." This is a simplistic view of the potential role of VLDL in the interplay of cells, coagulation proteins, and the regulatory systems involved in vivo. To realize the degree of complexity that we may need to address, we need only look at the work of Booyse et al in this issue of Seminars, in which they demonstrate that hypertriglyceridemic VLDL, in contrast to normal VLDL, do not support the early release of t-PA from endothelial cells, an antifibrinolytic event.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3051391 TI - Platelet lipoprotein interactions. PMID- 3051392 TI - Effects of omega-3 fatty acids in hypertriglyceridemic states. AB - In the experimental studies reported in this review, dietary omega-3 fatty acids from fish and fish oil had profound hypolipidemic effects in normal subjects and in hypertriglyceridemic patients with combined hyperlipidemia (type IIb) and type V hyperlipidemia. In these studies, 68 adults participated in carefully controlled metabolic experiments. In all subjects and patients, there were marked reductions in plasma cholesterol and triglyceride concentrations, with triglyceride lowering being especially great. There were also reductions in VLDL, chylomicrons, remnants, LDL, apo B, and apo E. The HDL changes were inconstant and varied from subject to subject. Whereas the mechanism of the hypolipidemic action of the omega-6-rich vegetable oils containing linoleic acid, such as corn or safflower oil, still remains obscure, the mechanism of action of the omega-3 fatty acids in fish oil has been well documented within a few years of their use as hypolipidemic agents. The synthesis of triglyceride and VLDL in the liver is greatly reduced by omega-3 fatty acids. At the same time, the turnover of VLDL in plasma is greatly shortened. LDL production is decreased. Combined with other dietary manipulations, such as a reduction in saturated fat and dietary cholesterol, the use of omega-3 fatty acids to treat hyperlipidemic and especially hypertriglyceridemic patients would appear to have a well-supported rationale. Further studies are required to delineate exact doses and precise indications for different types of hyperlipidemia and to differentiate the effects of, if any, the two major omega-3 fatty acids in fish oil, EPA and DHA. Coupled with the known antithrombotic actions of omega-3 fatty acids from fish oil because of changes in prostaglandin secretion and platelet function, these hypolipidemic effects would appear to have an important potential role in the control of coronary heart disease and other atherosclerotic disorders. PMID- 3051393 TI - Radionuclide evaluation of liver transplants. AB - Orthotopic liver transplantation is now an established technique for treating patients with various forms of end stage liver disease. The number of centers performing the procedure is increasing and, as the number of transplant recipients in the population increases, many institutions performing nuclear medicine studies will be confronted with requests to evaluate these patients. While a variety of radionuclides are proving useful in this evaluation, the 99mTc iminodiacetic acid (IDA) compounds, particularly 99mTc diisopropyl IDA (DISIDA), will probably account for the majority of radionuclide evaluations of these patients because they are well suited to monitor both structural and functional changes of the graft. The primary application of radionuclide studies is focused in the postoperative period, when problems with the vascular and biliary anastomoses, rejection, infections, and bile leaks all produce alterations in radionuclide hepatobiliary studies. Abnormalities such as rejection and infection produce primarily functional, rather than structural changes and are not easily differentiated based upon the kinetics of 99mTc-DISIDA extraction and excretion by the liver, serial imaging and correlation with clinical data is necessary in such situations. Quantitative analyses of kinetic 99mTc IDA (DISIDA) studies and quantitative approaches with other compounds such as 99mTc galactosyl neoglycoalbumin (NGA) may permit better assessments of relatively subtle changes in liver function in the posttransplant period. PMID- 3051394 TI - The effects of denervation and acute rejection on left ventricular volumes measured by radionuclide ventriculography following cardiac transplantation in the chacma baboon. AB - Seven baboons underwent autotransplantation of the heart or heart and both lungs (group A). Eleven allografts were performed (group B) (nine orthotopic heart transplants and two en bloc transplants of the heart and both lungs). Radionuclide ventriculography was performed both pretransplant and at intervals posttransplant in all animals, and provided measurements of ejection fraction (EF) and left ventricular volumes (LVv) (end-diastolic volume [EDV], end-systolic volume [ESV], and stroke volume [SV]). In seven animals, a total of 20 endomyocardial biopsies were taken. Correlation was made between histopathological features of acute rejection seen on endomyocardial biopsy and changes in EF and LVv measured by radionuclide imaging. A significant increase of 12% in the EF (P less than 0.01) and significant falls in the LVv were observed in all animals (groups A and B) on the first posttransplant day, presumably a result of total cardiac denervation. EDV was reduced by 50% (P less than 0.005), ESV by 62% (P less than 0.0001), and SV by 43% (P less than 0.0001). In autografted baboons (group A) EF and LVv showed no further changes until reinnervation of the heart had occurred, when they reverted to pretransplant levels. In the allografted baboons (group B) further significant reductions in the LVv occurred as acute cardiac rejection progressed. From the first post transplant day to the time of the final study before the animals' death, the EF decreased by 10% (P less than 0.01), the EDV by 38% (P less than 0.005), and SV by 73% (P less than 0.003): the decrease in ESV did not reach statistical significance.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3051395 TI - The role of nuclear cardiology procedures in the evaluation of cardiac function following heart transplantation. AB - Heart transplantation is, today, an accepted and recommended modality in the management of selected patients suffering from terminal heart disease. However, acute rejection and infection remain the major complications of this operation. Serial endomyocardial biopsy (EB), considered as the standard for diagnosis of cardiac rejection, is an invasive and delicate operation, not free of complications, even when done by skilled personnel in specialized centers. The object of this study was to compare and correlate between radionuclide ventriculography (RNV) and the histologic findings of EB. Furthermore, to validate the use of nuclear cardiology techniques that allow noninvasive, reliable, and rapid quantitation of ventricular function and myocardial perfusion for the diagnosis and management of rejection in patients with heart transplants. Radionuclide studies of left ventricular function were performed in 3 heterotopic heart transplant patients (HHT) with long term survival and early after the operation in 5 patients with HHT, 12 orthotopic heart transplants (OHT) and in 2 heart and lung transplants (HLT). Simultaneous EBs were performed in the early posttransplant patients and a histologic score for acute rejection was obtained. First pass (FP) and multigated equilibrium blood pool ventriculography, using the in vivo 99mTc-labelling of RBCs was used to measure left ventricular volumes (LVV) such as stroke volume (SV), end-diastolic volume (EDV), end-systolic volume (ESV), and both global and regional ejection fraction (EF, REF). The histological grading of acute rejection was classified into four groups: (1) no rejection, (2) mild rejection, (3) moderate rejection, and (4) severe rejection. The median of each LVV parameter was calculated and correlated with the EB using a nonparametric one way analysis of variance. A percentage change of LVVs was used rather than the difference of the calculated LVVs. During moderate acute rejection, SV had the highest correlation in P less than 0.004, followed by the EDV (P less than 0.05), and finally ESV (P less than 0.02). During severe acute rejection the correlation was SV (P less than 0.0008), EDV (P less than 0.001), and ESV (P less than 0.006). Myocardial perfusion scintigraphy using 201T1 was performed in the HHT patients, although, at this stage we have not attempted a correlation with the histologic findings. In one patient with long term survival OHT, increased 131I-metaiodobenzylguanidine (MIBG) myocardial uptake was evident during a rejection episode. PMID- 3051396 TI - Evaluation of bone graft viability. AB - Radionuclide bone single photon emission computed tomography (SPECT) scintigraphy is an excellent method for the assessment of the vascular patency and bone viability. The scintigraphic findings are important in the management of microvascularized bone grafts used in the reconstruction of bone defects. These living bone grafts are necessary when the blood supply at the host site has been compromised as a result of radiation therapy or when the bone defect is greater than 6 cm. Bone scintigraphy is unique because the uptake of radiopharmaceutical is dependent both on an adequate delivery system and a living network of osteocytes. SPECT scintigraphy provides improved imaging characteristics and structural detail of the grafted bone. Eleven patients with microvascularized bone grafts to the mandible demonstrated marked bone reaction of the entire bone graft in the postoperative period. The accumulation of radiolabelled 99mTc methylene diphosphonate (MDP) correlated extremely well with postoperative findings and clinical response. SPECT bone scintigraphy was a safe, simple and effective method for the assessment of both patency of anastomosed blood vessels and the metabolic viability of the microvascularized bone graft. PMID- 3051397 TI - Assessment of the viability of skin grafts. AB - A number of tests are available to monitor the blood flow in free and distant pedicle skin grafts. The information from these tests aids in the development of measures to enhance vascularization and is occasionally needed to make clinical decisions in patients with distant pedicle grafts. Measurements of the disappearance of an intradermally injected small amount of 133Xe allows determination of a clearance rate and blood flow before and after clamping the original blood supply through the base. With 99mTc, which is generally more readily available, a flow index and block index can be determined. Clinically both procedures give equally good results in determining a safe time for pedicle base separation. The fluorescein test allows assessment of regional blood flow distribution within the pedicle. PMID- 3051399 TI - Principles of clinical trial design. PMID- 3051398 TI - An overview of clinical trials in oncology. PMID- 3051400 TI - Selection of patients for clinical trials. PMID- 3051401 TI - Quantitation of tumor response to anti-neoplastic therapy. PMID- 3051402 TI - Identification and assessment of prognostic factors. PMID- 3051403 TI - Meta-analysis: the quantitative approach to research review. PMID- 3051404 TI - Clinical trials in pediatric oncology. PMID- 3051405 TI - Marrow transplantation for hematologic malignancies: a brief review of current status and future prospects. AB - The ability to transplant bone marrow makes it possible to treat patients with hematologic malignancies with doses of systemic radiotherapy or chemotherapy that would ordinarily result in fatal myelosuppression. With this approach, some otherwise incurable patients can be treated successfully. For most hematologic malignancies (acute nonlymphocytic leukemia, acute lymphocytic leukemia, chronic myelogenous leukemia, and malignant lymphoma), if transplantation is delayed until patients have end-stage disease (drug-resistant relapse) or blast crisis, approximately 10 to 20% of patients can be saved. If transplantation is carried out earlier in the course of disease, the outcome improves considerably. Some of the major limitations of marrow transplantation are the need for an appropriate source of marrow, the severe myelosuppression seen in the immediate posttransplant period, the complications of graft-v-host disease (GVHD), and disease recurrence. The use of autologous marrow and the formation of a national donor registry may make transplantation more widely applicable. The availability of growth factors can hasten hematopoietic recovery, making the immediate posttransplant period safer. New immunosuppressive regimens and T-cell depletion of donor marrow can diminish the impact of GVHD. The need for better preparative regimens persists and has emerged as the major requirement for continued progress in marrow transplantation. PMID- 3051406 TI - Monoclonal antibodies in the treatment of hematopoietic malignancies. AB - The development of practical methods for producing specific, highly purified monoclonal antibodies (MoAbs) targeted to specific tumor-cell antigens has opened new vistas in therapy for both hematologic malignancies and solid tumors. Numerous phase I trials with first-generation MoAbs have revealed both the broad therapeutic promise of MoAbs in anticancer therapy and the obstacles that must be overcome before they can be utilized fully in the clinical setting. MoAbs can be used to activate or manipulate the patient's immune system to destroy malignant cells. They also can be used as carriers for antitumor agents such as chemotherapeutic compounds, radiation, and toxins. Because many MoAbs are prepared from mouse cells, a common problem has been the development of resistance to the heterologous mouse protein. Strategies to circumvent this resistance include the use of human MoAbs, the production of chimeric mouse-human MoAbs, and the induction of tolerance to the mouse MoAbs. The possibility of combination therapy using MoAbs in conjunction with chemotherapy, lymphokines, or other biological response modifiers also is being explored. PMID- 3051407 TI - Colony-stimulating factors: present status and future applications. AB - Research in recombinant DNA technology has led to the characterization of colony stimulating factors (CSFs) as a family of glycoprotein hormones that are thought to regulate blood cell proliferation and differentiation. CSFs also have been studied for their potential use in treating various hematologic, infectious, and neoplastic disorders. Preliminary-results using granulocyte-macrophage colony stimulating factor to restore leukocyte competence in acquired immune deficiency syndrome and myelodysplastic syndrome patients have been impressive. Toxic effects generally have not been severe. Other hematopoietic factors are receiving scrutiny for their potential clinical applications. PMID- 3051408 TI - Mechanistic aspects of hepatic bilirubin uptake. PMID- 3051409 TI - Analytic aspects and clinical interpretation of serum bilirubins. PMID- 3051411 TI - Kernicterus and bilirubin encephalopathy. PMID- 3051410 TI - Neonatal jaundice. PMID- 3051412 TI - Epidemiology of unconjugated hyperbilirubinemia: revisited. PMID- 3051413 TI - Postoperative jaundice. AB - Jaundice in the postoperative patient is a common complication of surgery that may be a confusing and potentially serious problem. The surgical patient is subjected to a variety of stresses, such as hypotension, infection, drugs, and anesthetic agents, many of which are potentially hepatotoxic. Management of the patient with postoperative jaundice can be difficult because the precise cause of the hepatic insult is frequently indeterminate. It is important for the clinician to recognize the patterns of liver injury that may occur in the postoperative period in order to initiate appropriate management. PMID- 3051414 TI - Case studies in jaundice of pregnancy. PMID- 3051415 TI - Paleoradiologic evaluation of the Egyptian royal mummies. AB - We examined radiographs of 12 Egyptian royal mummies obtained by two of the authors (W.R. and J.E.H.) and never before published. These radiographs demonstrate findings not previously described in Egyptian mummies, including congenital lunate-triquetral fusion and destructive skeletal lesions not explainable on the basis of vandalism by tomb robbers. Antemortem fractures, degenerative joint disease, and arterial vascular calcification were also seen. In 11 of the 12 cases, there was chondrocalcinosis of intervertebral discs or menisci, probably an artifact of embalming. Visceral packing and skeletal deformity due to wrapping were observed, as well. Radiology provides important paleopathologic and archeologic information for the accurate, comprehensive study of Egyptian mummies. PMID- 3051416 TI - Poverty and disease: need for structural change. AB - In underdeveloped countries, more than 70% of the population suffers from infectious and communicable diseases. These diseases are transmitted with the help of poor water, sanitation, housing, etc. Further, education and nutrition also affect the vulnerability of the individual. All these factors responsible for disease, are themselves dependent on income--the lower the income, the lower education and nutritional status, and the poorer quality the water, and housing. An increase in the absolute income for some, and a redistribution of income for all, are necessary to cure the ills of society. It is possible to increase the real income of individuals by giving them hand-outs or dealing with the problems of their 'basic needs'. However, these approaches do not take into account the underlying factors responsible for disease, and are severely limited in scope. The elite in a country, who, to a great extent, determine the role of the State and of the government, are only willing to give a certain amount of charity, and nothing more. They will, clearly never give to the poor, so much that their own (relative and absolute) position is threatened. The only way possible is through a government which works for the majority of the people, rather than for a small elite. PMID- 3051417 TI - From vice to madness: the semantics of naturalistic and personalistic understandings in Trinidadian local medicine. AB - Whilst the common analytical distinction between 'naturalistic' and 'personalistic' paradigms of medical knowledge has some immediate heuristic value and may indeed closely resemble the explicit schema elaborated by informants, interpretation of the semantics of bush medicine and madness in Creole Trinidad suggests that the two types of knowledge are not incompatible, nor mutually exclusive, nor distinct. The vices of ganja and rum use may be interpreted within the hot-cold classification of bush medicine but, like other vices and like the more 'intrapersonal' categories of pressure, grinding, studiation and tabanka, they may be understood as leading to madness. A common idiom of opposition and catharsis unites them, providing a higher level of analytical generality manifest in a range of local social institutions, and one rooted in post-colonial Afro Caribbean experience and ideology. PMID- 3051418 TI - Cancer care--a stress for health professionals. AB - Literature related to health care professionals dealing with stress of cancer care is still in its infancy. The authors distinguish papers of general interest (the most frequent), papers identifying stressors, and papers about stress consequences. Most of them recognize death of the patients as a major stressor for health care professionals. There are also additional stressors specific to health care and work. Consequences of stressors have another important dimension: working with cancer patients is often a chronic stress which may lead to the development of burnout and poor quality of care. Little also is actually known about how coping strategies and/or support are influencing adaptation and stress consequences. The authors suggest that an important effort should be made to evaluate stress, and its consequences on poor staff communication with cancer patients and their families. Training interventions aimed at a better quality of care should be designed and their usefulness investigated. The effectiveness of training for health care professionals dealing with cancer patients is reviewed. PMID- 3051419 TI - Predictors of prenatal care utilization. AB - Despite substantial evidence linking improved pregnancy outcomes with receipt of prenatal care and recent improvements in prenatal care utilization, specific subpopulations continue to receive inadequate or less than adequate care. The study reported here examined the predictive power of a set of variables describing the type of financial coverage available to the mother, attributes of the mother, father and family and characteristics of the health care system. A stratified random sample of mothers was generated from state birth certificate files and surveyed through the use of a mailed questionnaire. Stratification was designed to assure adequate representation of subgroups expected to receive less adequate prenatal care. The study findings indicate that there were deficiencies in prenatal care utilization and that these deficiencies were concentrated in specific areas and subpopulations within the state. While the majority of women in the study started prenatal care in the recommended first trimester, most did not maintain the recommended schedule of visits with their care provider. The following conditions were found to reduce the likelihood of receiving adequate care after controlling for service need: younger women (particularly adolescents); less educated (particularly those without a high school education); low income; longer travel time; Medicaid recipient; and rural resident. In addition, it was found that where one lives is a significant predictor of the adequacy of prenatal care even after controlling for all of the above variables. The authors conclude that it is important in assessing potential policy and program options for reducing differentials in prenatal care use to distinguish between economic and noneconomic barriers to utilization. Receipt of Medicaid does not assure adequate prenatal care use.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3051420 TI - The development of medical sociology in Finland. AB - This paper reviews the development and major directions of medical sociology in Finland. An interest in the social aspects of health can be traced to 19th century social medicine. A general interest in medical sociology emerged in the 1950s but the research was mainly focused on the work setting and behavior of various health professionals. In the 1960s, the regional differences in morbidity and accessibility to health care were the focus of the research sponsored by the government, and many studies examined the health behavior and health status of selected groups. In the 1980s, the research has covered the most topical areas in the field. Although there are a large number of active researchers, the field is characterized by a weak institutionalization. PMID- 3051421 TI - Climate, diffused solar radiation and multiple sclerosis. AB - Environmental factors implicated as affecting world-wide distribution of multiple sclerosis are reviewed. It is suggested that climate may be involved in the etiology of this disease. Diffused solar radiation effects on multiple sclerosis hospital admission rates are demonstrated. PMID- 3051422 TI - HMOs in the U.S.A. and Britain: a new prospect for health care? AB - Health Maintenance Organisations (HMO) are poised to become the mainstream form of health care delivery in the U.S.A. by the end of the next decade. This article charts their progress and the expectations held for them in both the U.S.A. and Britain. It is argued that HMOs are undergoing corporatization in the U.S.A., while in the U.K. they are being considered chiefly as instruments of privatisation and as a method for introducing managerial authority into the medical profession. The claims of advocates of HMOs, in terms of their competitive effects, reduced costs and enhanced quality of service, are reviewed. Although no HMOs exist in Britain, they have found support in influential quarters. The proposals and claims of the advocates of HMOs in Britain are reviewed. Although HMOs have not yet found favour with the current government, it is argued that the concept will continue to receive support and may become a component of health policy in the near future. PMID- 3051423 TI - A review of the social patterning and significance of measures of height, weight, blood pressure and respiratory function. AB - This paper reviews evidence about the social patterning and relationship to life chances of four measures of health, development or functioning; namely height, weight, blood pressure and respiratory function. It argues that these are useful supplements to more commonly used measures of 'health' such as mortality rates, morbidity rates, or self reports of health, and recommends the increased use of such direct physical measures in routine statistics and special health surveys. PMID- 3051424 TI - Defining death: organ transplants, tradition and technology in Japan. AB - This article explores Japanese attitudes about brain death and organ transplantation. First, ancient burial customs and death-related rituals associated with Shinto and Buddhism are examined. Next, contemporary attitudes towards the dead are discussed in the context of current controversies surrounding brain death and organ transplantation. Finally, an attempt is made to link the traditional Japanese views of death with modern medical dilemmas. PMID- 3051425 TI - Clinical efficacy of temporal artery biopsy in Nashville, Tennessee. AB - A retrospective study of temporal artery biopsies done over one decade in four Nashville hospitals yielded 412 procedures on 394 patients. Diagnosis was obtained in 17%, and the procedure was helpful in 21% of patients. Only two complications were recorded. There was no correlation between length of biopsy specimen and diagnostic yield. The Nashville experience with temporal artery biopsy is probably representative. Some refinement of procedure application appears necessary. PMID- 3051427 TI - Noninvasive evaluation of ischemic stroke with SPECT. AB - Technetium Tc 99m DTPA single photon emission computerized tomography (SPECT) brain scans of 20 patients with acute ischemic stroke were reviewed retrospectively and compared with clinical and radiologic (CT) data. Fourteen of the patients had abnormal SPECT studies. The abnormal findings were demonstrated by static views in eight patients, by the flow study in one patient, and by both sets of images in the other five patients. All abnormalities correlated with the clinical syndrome of presentation, and only two of the patients had no corresponding lesions on CT. Of the six patients with normal SPECT scans, two had abnormal CT studies, and in the other four, no lesions were shown at all. The ability of 99mTc DTPA SPECT to display cerebral infarctions appears to be, at best, comparable to that of CT. SPECT also provides qualitative information regarding flow dynamics in the affected hemisphere of some patients (6/20 in our review). This, we believe, represents the objective demonstration of the preexisting insufficient collateral flow in the hemisphere at risk for ischemic stroke. PMID- 3051426 TI - Intracranial tuberculoma in children: a new look at an old problem. AB - Intracranial tuberculoma has become a rare cause of space-occupying intracranial lesions in childhood, but it must still be considered in the differential diagnosis. Tuberculosis remains a significant disease in developing countries and in the United States, and tuberculoma is a well known presentation of childhood tuberculosis. This diagnosis must be considered especially in persons traveling or living in developing countries and in immigrants from third-world areas. We report three cases of tuberculoma in children seen during one year at our institutions to illustrate the need for continued suspicion. We summarize the clinical presentation and current treatment recommendations and review the available literature. PMID- 3051428 TI - Headache after lumbar puncture: review of the epidural blood patch. AB - The incidence of accidental dural puncture during epidural block is 2.9%; headache follows in as many as 76.5% of these patients. Treatment by injection of autologous blood into the epidural space has gained wide acceptance since its introduction in 1960, though it is contraindicated by blood dyscrasias, anticoagulant therapy, bleeding, and localized infection. The procedure is done by slowly injecting 15 to 20 ml of blood into the same interspace, no sooner than 24 hours after the original puncture. Serious complications are rare. PMID- 3051429 TI - Pancreatic heterotopia: a reappraisal and clinicopathologic analysis of 32 cases. AB - A total of 32 histologically documented cases of heterotopic pancreas was found in a review of the records of the department of pathology at the Chang Gung Memorial Hospital between 1977 and 1987. This review was done to ascertain the clinical significance of this uncommon entity. In 14 patients (44%), the aberrant pancreatic tissue was symptomatic; in the other 18 (56%), it was found incidentally. In the symptomatic group, the heterotopic pancreatic tissue was found in a duplication cyst of the ileum in one patient, in the common bile duct in one, in a Meckel's diverticulum in four, in the stomach in three, in a congenital duodenal diaphragm in one, in the duodenum in three, and in the ileum in one. The majority of heterotopic pancreatic tissue in the asymptomatic group was encountered in the jejunum (15 patients). Symptoms were related to complications, including obstruction of the common bile duct, mucosal ulcer with hemorrhage, intussusception, and intestinal obstruction, but not to pathologic conditions of the pancreas itself, such as pancreatitis or pancreatic cyst or neoplasm. In all of the clinically significant cases, the clinical symptoms disappeared completely after surgical removal of the aberrant tissue. In 28 cases (87%), diagnosis was made by frozen section during operation. Preoperative diagnosis of aberrant pancreas was not made in any of the cases. Histologically, all cases showed pancreatic excretory ducts; in 31 cases (97%), exocrine glands were present, and in 27 cases (84%), islets of Langerhans were discernible. There was no relationship between symptoms and the presence of islets, acini, or ducts. Mallory's phosphotungstic acid-hematoxylin stain was used to demonstrate zymogen granules in the acinar cells, and insulin, glucagon, and somatostatin were demonstrated with the horseradish peroxidase-antihorseradish peroxidase immunocytochemical staining technique; islets of Langerhans were also identified. Technetium Tc 99m scintigraphy was used to detect the bleeding source in a Meckel's diverticulum and an enteric duplication associated with ectopic gastric mucosa. PMID- 3051430 TI - Thrombophilia: new ideas about old problems. AB - Patients with recurrent thrombosis may have one of the thrombophilias described in recent literature. These clotting disorders may involve abnormalities in platelets, fibrinogen, plasminogen, protein C, protein S, or antithrombin III. There is usually a family history of clotting disorders, since most of these are inherited in an autosomal dominant pattern. PMID- 3051431 TI - The physician's obligation to treat AIDS patients. AB - Medicine's triumph over contagious disease through improved techniques of prevention and treatment in the decades before the appearance of the acquired immunodeficiency syndrome (AIDS) left physicians with little impetus to explore their feelings regarding the acceptance of personal risk in the course of patient care. The rapid expansion of the AIDS epidemic, however, has made it essential for every physician and medical student to confront this issue and determine whether he is willing to accept the minimal risks of transmission posed by the human immunodeficiency virus (HIV) to health care workers. This paper will present five arguments in support of the contention that the physician is obligated to treat all those who would benefit from his care, even when such care entails personal risk to himself. These arguments include the historical traditions of the profession, formal ethical codes, the dependent nature of the patient, the social contract, and medicine as a profession. PMID- 3051432 TI - Computer-assisted learning compared with weekly seminars for teaching fundamental electrocardiography to junior medical students. AB - This study was designed to assess whether a self-study interactive computer program is more effective than weekly seminars for teaching fundamental skills of electrocardiographic interpretation to junior medical students. Forty-two students were assigned to the computer and 41 to the seminar group. A test was given to each participant at the beginning and end of each rotation. The computer group used a computer-assisted learning program, and the seminar group met weekly with a cardiologist to review electrocardiograms. Attendance at a minimum of 80% of the seminars or completion of 80% of the computer-assisted learning program was required for inclusion in the statistical analysis. The mean difference in test scores before and after study was 5.69 for the computer group and 4.36 for the seminar group (P less than .02 by one-tailed t-test). These results indicate that the computer group performed significantly better than the seminar group. We believe this difference to be educationally important. PMID- 3051433 TI - Marshall-Smith syndrome: further delineation. AB - We have reported a case of the Marshall-Smith syndrome, a condition characterized by accelerated bone maturation, dysmorphic features, respiratory compromise, failure to thrive, neuro-developmental abnormalities, and death in early infancy. Early evaluation of upper airway obstruction in these children may be important in prolonging life. Although the prognosis has been poor to date, early recognition and aggressive multi-disciplinary therapy may permit further investigation into the cause and appropriate management of this disorder. PMID- 3051434 TI - Unicameral bone cyst of the tibia complicated by genu valgum. AB - Progressive tibia valga occurred in an adolescent girl after curettage and allografting of a large unicameral bone cyst of the tibia. The valgus deformity was corrected by a closing-wedge osteotomy, which resulted in a good functional and cosmetic result. The progressive valgus deformity might have been caused by stimulation of overgrowth of the medial tibial metaphysis. PMID- 3051435 TI - Necrotizing fasciitis in a renal transplant patient. AB - We have described a 28-year-old diabetic woman who had necrotizing fasciitis of the perineum three years after receiving a living related renal transplant. The diagnosis of necrotizing fasciitis was made early and she was referred to a tertiary care center where she received radical perineal debridement and aggressive medical and surgical follow-up. Necrotizing fasciitis in a transplant patient is rare; review of the literature shows few cases and no survivors. Our patient has returned to a normal life despite continuation of all immunosuppressive therapy throughout the entire hospital course. In addition, she had a good cosmetic result despite the large necrotic perineal infection. Her survival can be attributed to early diagnosis and referral, immediate and extensive debridement, and aggressive protein replacement. PMID- 3051436 TI - Primary extraosseous osteogenic sarcoma of the mediastinum: clinical, pathologic, and radiologic correlation. AB - We have presented a rare case of primary involvement of the anterior mediastinum by osteogenic sarcoma in which CT was useful in demonstrating calcium within the mass, and in distinguishing the mass from surrounding bony structures. Although rare, extraosseous osteogenic sarcoma should be included in the differential diagnosis of a calcified anterior mediastinal mass. PMID- 3051437 TI - Microangiopathic hemolytic anemia associated with metastatic carcinoma of the colon. AB - We have reported a case of microangiopathic hemolytic anemia and metastatic carcinoma of the colon in a 38-year-old woman. Despite the mucin content and the high incidence of carcinoma of the colon in western populations, there are only four cases of colon cancer among the 77 reported cases of malignancy-associated MHA. PMID- 3051438 TI - Pulmonary hemorrhage and glomerulonephritis in the absence of anti-glomerular basement membrane antibody. AB - We have described a case of pulmonary hemorrhage and idiopathic crescentic glomerulonephritis unrelated to anti-glomerular basement membrane antibodies and/or immune complexes. Although pulmonary hemorrhage was controlled dramatically with high doses of corticosteroids, renal function declined rapidly. PMID- 3051439 TI - A review of the literature related to trunk muscle performance. AB - In the past, there has been no comprehensive review of the literature pertaining to different methods of assessing trunk muscle strength. This review describes the different studies that have been performed, and determines the hierarchy of strength values and agonist/antagonist strength ratios for the trunk musculature. In general, the strength hierarchy consists of, from strongest to weakest: extension, flexion, side bending, and rotation. The agonist/antagonist ratio for extension/flexion is 1.30 and for rotation and side bending, motion to the right approximately equals motion to the left. Changes in the relative strengths of the different trunk muscle groups is affected by spinal pathology, and this is discussed. Possible clinical implications and direction for future research are delineated based on the findings of this review. PMID- 3051440 TI - [Quality assurance in sonography]. PMID- 3051441 TI - [Use of orgotein in the treatment of plastic penile induration]. PMID- 3051443 TI - [Renal liposarcoma. Description of a case]. PMID- 3051442 TI - [A rare case of association of polycystic kidney and hypernephroma]. PMID- 3051444 TI - [Long-term captopril therapy of 3 sisters with Bartter's syndrome]. PMID- 3051445 TI - [Adverse effects of the use of cyclosporin A in patients having undergone a kidney transplant]. PMID- 3051446 TI - [Sympathetic neuroblastoma in adults]. PMID- 3051448 TI - [Biology of von-Willebrand factor]. PMID- 3051447 TI - [Epidermoid cysts of the testis. Considerations on a clinical case]. PMID- 3051449 TI - [Acute typhlitis in acute leukemia in an adult]. PMID- 3051450 TI - Surgical correction of renovascular hypertension. AB - The role of surgical revascularization in the management of patients with renal artery disease has changed in recent years. This has occurred owing to the advent of transluminal angioplasty as an effective method of treatment for certain patients, improved results of surgical revascularization in older patients with atherosclerosis, an enhanced appreciation of advanced atherosclerotic renal artery disease as a correctable cause of renal failure, and the development of more effective surgical techniques for patients with severe aortic atherosclerosis and branch renal artery disease. Surgical revascularization is at present the treatment of choice for patients with branch renal artery disease, ostial atherosclerotic renal artery disease, a renal artery aneurysm, and patients in whom renal angioplasty has been unsuccessful. Excellent clinical results continue to be achieved with surgical revascularization in properly selected patients. PMID- 3051451 TI - Surgery of adrenal disorders. AB - The surgical management of patients with adrenal disease depends on the functional nature of the endocrinopathy. It requires a thorough knowledge of adrenal anatomy and command of the various surgical approaches to the gland. Included for discussion are Cushing's syndrome, adrenal adenoma and adenocarcinoma, adrenocortical carcinoma, primary aldosteronism, and pheochromocytoma. PMID- 3051452 TI - Pelvic fracture and injury to the lower urinary tract. AB - The presence of a urologic injury must be considered in all patients with pelvic fracture. Uroradiographic evaluation starting with retrograde urethrography is indicated in all male patients with concomitant gross hematuria, bloody urethral discharge, scrotal or perineal ecchymosis, a nonpalpable prostate on rectal examination, or an inability to urinate. If the urethra is normal, a catheter may be passed, and in the presence of gross hematuria, a cystogram must be performed. Female patients rarely suffer urethral lacerations. The urethra is examined, and a Foley catheter may be passed without a urethrogram. The immediate management of associated urologic injuries continues to evolve and evoke controversy. Selected cases of extraperitoneal bladder perforation may be safely managed solely by catheter drainage. Intraperitoneal perforations require surgical exploration and repair. Urethral disruption (partial or complete) may be safely managed by primary cystostomy drainage with management of potential complications (stricture, impotence, incontinence) in 4 to 6 months. PMID- 3051453 TI - Etiology, classification, and management of renal trauma. AB - Management of blunt renal trauma demands an aggressive effort to define the extent and severity of the renal injury with imaging studies. In general, a conservative approach to treatment is recommended that may include an early surgical exploration when the risk of late hemorrhage is great and the kidney or a portion of the kidney has obviously already been lost. To treat all patients with surgery or with expectant treatment is illogical. If expectant treatment is elected and the patient has a significant renal injury, every effort should be made to follow the patient adequately with ultrasound or CT scans in order to identify at the earliest opportunity an expanding hematoma and prevent needless nephrectomy and shock. Of most importance is to avoid inadequate studies that fail to define the source of injury and lead in the long run to inadequate surgical management. A tongue-in-cheek representation of such a scheme of treatment is illustrated in Figure 5. PMID- 3051454 TI - Changing surgical aspects of urinary stone disease. AB - Surgical management of urinary calculous disease has changed dramatically in the past decade. The development of percutaneous nephrostomy techniques has allowed new access to upper tract stones. Percutaneous removal of large calculi was made possible by the development of ultrasonic and electrohydraulic lithotripsy. All upper tract calculi can now be removed in 70 to 100 per cent of cases with minimal complications. Nephrostolithotomy has reduced transfusion rates and hospitalization costs and has markedly shortened convalescence periods compared with open surgery. Ureteroscopy followed nephrostolithotomy as advanced fiberoptic technology allowed the development of the small-caliber instruments required for this procedure. With experience, successful stone retrieval has occurred in 90 per cent or more of cases, again with minimal complications. As nephrostolithotomy and ureteroscopy have become available, the subspecialty of endourology has emerged and significantly changed the management of urinary tract calculi. Perhaps the most significant advance in stone therapy has been the design and implementation of extracorporeal shock wave lithotripsy. With this noninvasive technique, most renal and proximal ureteral calculi can be effectively treated with minimal morbidity and convalescence. Research in lithotripter design is continuing, with more advanced and effective machines on the horizon. The applicability of extracorporeal therapy for the treatment of biliary tract calculi is currently under investigation. Finally, one should not disparage medical therapy for recurrent nephrolithiasis. A comprehensive metabolic evaluation combined with selective medical therapy provides almost complete relief from recurrent stone formation and makes medical therapy an integral component of treating the patient with renal or ureteral calculi. PMID- 3051456 TI - Nosocomial urinary tract infections. AB - Convenience to the hospital staff is certainly not an acceptable reason for the use of a potentially dangerous drainage tube. An indwelling urinary drainage catheter should be used only in patients who need multiple straight urinary catheterizations, develop urinary obstruction or incontinence, or are comatose and require frequent urinary output measurements. An indwelling catheter may also be needed for drainage or stenting during or following genitourinary surgery. Once it has been determined that urinary catheterization is necessary, a closed urinary drainage system catheter must be carefully and aseptically inserted by experienced hospital personnel after careful preparation. The closed drainage system must be meticulously maintained throughout the patient's hospitalization and catheterization. After the catheter is removed, a urinary culture should be performed to identify any postcatheter infection. If there is infection, the patient must be treated with antibiotics. If symptoms of a urinary tract infection, bacteremia, or sepsis ensue, treatment must be rapidly begun with antibiotics as appropriate on the basis of drug sensitivity testing. These techniques will not eliminate bacteriuria associated with urinary drainage catheters. However, they will reduce the incidence, morbidity, and mortality associated with urinary catheterization. PMID- 3051457 TI - Continent urinary reservoirs. AB - Public awareness of the alternatives to wet stomas and the patient's concern for body image have significantly increased the demand for continent urinary reservoirs. In the final analysis, preservation of renal function is the standard for judging the success of a diversion, and long-term observations must be made with this point in mind. It seems likely that the continent reservoirs now available will preserve renal function. PMID- 3051458 TI - Surgical management of urinary incontinence in children. AB - The field of continent reconstruction continues to expand rapidly, as new innovations are introduced by imaginative surgeons. Today, review of previous experience and knowledge of physiology permit creation of solutions to previously insoluble problems. It must be stressed that long-term results are not available for many of these procedures, but with careful follow-up, the outlook is promising. PMID- 3051455 TI - Controversies in urologic oncology. AB - The persistent controversies that characterize urologic oncology reflect the significant advances that have been made in our understanding of genitourinary malignancy and the objective posture that has been taken both by those active in this area and by those whose interests are more indirect. Ultimately, the beneficiaries of these controversies are the patients and their physicians. As continued problems are recognized and investigations are designed to explore them, controversy and skepticism become a healthy concomitant so that true knowledge and understanding can be achieved and such issues ultimately resolved. PMID- 3051460 TI - Evaluation and management of erectile dysfunction. AB - The man who seeks treatment for impotence typically has a sense of inadequacy that extends beyond sexual satisfaction to his business and personal relationships. Although no magic potion is available, current medical technology allows the surgeon to accommodate each patient who seeks treatment, guaranteeing sufficient penile rigidity to effect coitus. PMID- 3051459 TI - Urinary incontinence in adults. AB - Abnormalities of the micturition cycle are responsible for a large number of visits to the urologist. Some of these abnormalities produce annoying symptoms; others may lead to hazardous and disabling changes in the urinary tract. Urinary incontinence in adults results from an abnormality of one or more of the factors involved in the two phases of the micturition cycle: filling-storage and emptying. Proper management requires judicious selection among the pharmacologic and surgical alternatives. PMID- 3051461 TI - Effect of respiratory cycle on measurements of cardiac output by thermodilution. AB - Current recommendations for obtaining reproducible measurements of cardiac output by thermodilution are contradictory. Several investigators have shown that reproducibility of measurements can be improved by initiating injections at specific moments in the respiratory cycle. We prospectively studied 31 patients, comparing three thermodilution measurements of cardiac output at peak inspiration, end-exhalation and random in mechanically ventilated patients. Patients were separated into two groups: group 1 was composed of intubated patients on intermittent mandatory ventilation (IMV) without spontaneous overbreathing, and group 2 included patients on IMV with spontaneous overbreathing. The results from this study demonstrated no statistical difference in the ranges or standard deviations of the measurements of cardiac output among injections initiated at random, peak-inspiration or end-exhalation in either group. We conclude that the reproducibility of measurements cannot be enhanced by initiating injections at specific moments in the respiratory cycle. PMID- 3051462 TI - Choledochal cysts. PMID- 3051463 TI - Evaluation of endoscopic hemostasis using an improved clipping apparatus. AB - Endoscopic clipping hemostasis (ECH) is an effective method to control bleeding. However, ordinary clipping apparatuses have not been widely adopted because of several disadvantages. Accordingly, we developed an improved ECH apparatus that can be used during a general endoscopic examination without requiring an assistant. It is not only easy to operate but also ensures safe and effective hemostasis. The ECH apparatus was employed in 80 patients between February 1983 and August 1987 at the Sakura National Hospital. Fifty-one of the patients had upper gastrointestinal bleeding; in 29, preventive clipping was performed after polypectomy. Permanent hemostasis was maintained in 43 (84.3%) of the patients with upper gastrointestinal bleeding, and no bleeding was recognized in any of the 29 patients treated with prophylactic clipping following polypectomy. PMID- 3051464 TI - Acute abdominal pain. Actual surgical aspects of sonography. AB - The results of a prospective study show the most common causes of acute abdominal pain, and the diagnostic role of sonography in such cases is evaluated. Sonography performs three significant functions: visualization of pathologically changed organs, identification of healthy organs, and ultrasound-guided puncture of intra-abdominal collections of fluid. Affections in the right upper quadrant can be best diagnosed sonographically. Edematous pancreatitis can be easily identified; interpretation is difficult, however, in the presence of necrosis and abscesses. Therefore, computed tomography is also necessary. Sonography can differentiate between a mechanical and a paralytic ileus, but radiological examination is still necessary. In inflammatory diseases of the gastrointestinal tract, sonography may be helpful for diagnosis. An aortic aneurysm can be easily identified, whereas mesenteric infarction cannot. In the future, however, the use of a duplex scan may help in recognizing this condition. Affections of the abdominal wall can be visualized well by using a high-frequency transducer. PMID- 3051465 TI - Retinopathy of prematurity. AB - Over the last decade major advances have been made in the understanding of the pathogenesis and evolution of retinopathy of prematurity (ROP). The increased survival of very small premature infants in modern neonatal intensive care units has led to the resurgence of this potentially blinding disease. ROP appears to be a multifactorial disease, the prevention of which is probably impossible even now, with the most accurate methods of blood gas monitoring and oxygen restriction. In addition to oxygen, there are a number of significant risk factors, such as birth weight and gestational age, ventilator hours, hyper and hypocarbia, hypoxia and acidosis, xanthine therapy and probably bright light. Current data suggest that the level of antioxidants in the immature retina is relatively low and therefore oxygen radicals which accumulate in the preterm baby's retina may play an important role in the pathogenesis of ROP. The treatment of the disease in both its "active" and "cicatricial" stages emphasizes the need for a new classification which could serve as a common international language through which results may be compared. Vitamin E was suggested in some studies to be helpful in preventing the severe stages of the disease, but its efficacy has yet to be proved. Treatment modalities such as photocoagulation, cryotherapy and vitrectomy are being tried as a means of therapy in the more advanced stages of the disease. Preliminary results of a large multicenter study support the efficacy of cryotherapy. PMID- 3051466 TI - Choroidal osteoma. AB - This review details the characteristic clinical features, diagnostic approaches, management, and prognosis of the choroidal osteoma. A comprehensive differential diagnosis is organized to help the ophthalmologist differentiate this tumor from conditions which can sometimes be clinically similar, such as amelanotic choroidal melanoma and nevus, metastatic choroidal carcinoma, choroidal hemangioma, metastatic and dystrophic intraocular calcification, and others. The pathology of the choroidal osteoma is illustrated and several theories of the pathogenesis of this tumor including the possibilities of a choristomatous, inflammatory, and hereditary etiology are discussed. The recently recommended therapeutic technique of photocoagulation of subretinal neovascularization associated with choroidal osteoma are discussed. PMID- 3051467 TI - Corneal hypoesthesia. AB - Decreased corneal sensitivity is an important clinical parameter in neuro ophthalmological evaluation. It may be associated with such disorders as diabetes, Herpes simplex keratitis, and myasthenia gravis, with corneal toxicity via chemical exposure or ocular drug therapy, and with ocular surgery. In this review, methods and precautions involved in measuring corneal sensitivity are summarized, and the clinical conditions in which corneal hypoesthesia may exist are discussed. PMID- 3051468 TI - Pingueculae and pterygia. AB - Pingueculae and pterygia are benign peribulbar lesions composed of degenerated basophilic subepithelial tissue. Pingueculae do not affect vision, and minor irritation can usually be managed with artificial tears. Pterygia may affect the visual axis and require surgical and adjunctive treatment. The various therapeutic strategies are reviewed. A conservative approach is advocated, as surgical removal of primary pterygia may result in recurrent ptergyia that are more difficult to manage than the primary lesions. PMID- 3051469 TI - Invasive squamous cell carcinoma of the conjunctiva presenting as necrotizing scleritis with scleral perforation and uveal prolapse. AB - A 64-year-old white man presented with necrotizing scleritis with scleral perforation and uveal prolapse. Pathologic examination revealed squamous cell carcinoma of the conjunctiva invading adjacent corneal stroma and ciliary body. Invasive squamous cell carcinoma of the conjunctiva is uncommon, and intraocular invasion has rarely been reported in the literature. PMID- 3051470 TI - Cadaveric renal transplantation in the cyclosporine and OKT3 eras. AB - With advances in clinical immunosuppression, results in organ transplantation continue to improve. During a 52-month period, 507 cadaver renal transplants were performed, including 435 primary and 72 nonprimary transplants. All patients were managed with quadruple immunosuppression (prednisone, azathioprine, sequential MALG and cyclosporine). Our experience is divided into pre-OKT3 (n = 228) and OKT3 (n = 279) eras. All kidneys were harvested locally and preserved with pulsatile machine perfusion. The mean duration of preservation was 30.1 hours, with an organ utilization rate of 98.1%. The preservation-related dialysis rate was 13.6%, and primary nonfunction occurred in 8 kidneys (1.6%). Actuarial patient survival in primary and secondary transplant recipients was 90% at 3 years. Overall primary graft survival was 81.6% and nonprimary graft survival, 61.1%. However, the current OKT3 era is characterized by improved patient survival (98% vs 90%, p = 0.001) and primary graft survival (91% vs 80%, p = 0.002) at 1 year when compared with the previous era. Forty-nine patients have received OKT3 therapy, with 31 grafts (63.3%) successfully rescued. Cadaveric renal transplantation with machine preservation, quadruple therapy, and OKT3 rescue is associated with excellent early graft function, reduced acute rejection, and improved patient and allograft survival, even in high-risk recipients. PMID- 3051471 TI - Acute colonic ileus (pseudo-obstruction) in renal transplant recipients. AB - Colon complications are a potential source of serious morbidity to the immunosuppressed patient. Because of multiple predisposing factors, renal transplant patients are a high-risk group for the development of acute colonic pseudo-obstruction. During a recent 18-month period, 290 renal transplants (79 living, 211 cadaveric donors) were performed and prospectively analyzed for colonic dysmotility. A total of 34 episodes of acute colonic ileus (30 primary, 4 recurrent) occurred in 30 (10.3%) renal transplant recipients. Acute colonic ileus was more frequent after living-donor transplantation (19.0% vs. 7.1%, p = 0.006). Analysis of multiple variables revealed that the incidence of acute colonic ileus was directly related to mean cumulative prednisone dosage (p less than 0.05). Medical therapy (rapid steroid reduction, bowel rest) resulted in a 76.7% response, whereas 8 patients underwent colonoscopy because of progression to acute pseudo-obstruction. The success rate for colonoscopic decompression was 87.5%; in 1 patient cecal perforation developed after unsuccessful decompression. Overall, 33 of 34 (97.1%) episodes of acute colonic ileus were successfully treated. Steroid-induced ileus (pseudo-obstruction) is a potentially malignant early form of colonic dysmotility infrequently reported in transplant recipients. Successful management requires early clinical recognition, reduction in steroid dosage, bowel rest, and urgent colonoscopic decompression in select cases. PMID- 3051472 TI - Renal autotransplantation: an alternative to standard renal revascularization procedures. AB - From January 1978 through December 1987, 22 patients underwent 23 renal autotransplantation procedures for the treatment of renovascular hypertension through the retroperitoneal approach. The causes of the renal artery stenosis were as follows: atherosclerosis (15), fibromuscular dysplasia (6), and Takayasu's arteritis (1). Indications for renal autotransplantation were as follow: disease extending into the renal artery branches (10), stenosis of multiple renal arteries (6), atherosclerotic aorta in high-risk patients (4), and stenosis of renal artery in children (2). The mean preoperative blood pressure of 205 +/- 6/109 +/- 3 mm Hg decreased significantly to 139 +/- 4/77 +/- 2 mm Hg (p less than 0.001). The serum creatinine decreased significantly from a mean preoperative level of 2.2 +/- 0.8 mg/dl to a mean postoperative level of 1.4 +/- 0.4 mg/dl (p less than 0.05). Eleven patients with preoperative renal dysfunction had a significant decrease in the serum creatinine from a mean preoperative level of 3.4 +/- 0.3 mg/dl to a mean postoperative level of 1.9 +/- 0.2 mg/dl (p less than 0.001). One operative death occurred as a result of myocardial infarction. There were three postoperative complications, none of which affected the ultimate result in blood pressure or renal function. This experience demonstrates that in selected patients, renal autotransplantation is an excellent alternative in the surgical treatment of renovascular hypertension. PMID- 3051473 TI - Ultrasonographic features of carotid plaque and the risk of subsequent neurologic deficits. AB - In a prospective study, 214 consecutive patients considered not to be candidates for surgical intervention were evaluated by means of duplex scanning. Of the patients, 135 had no symptoms and 79 had a history of previous neurologic symptoms. In 139 sides duplex scanning demonstrated nonhemodynamic stenosis (lumen diameter reduction, less than 50%) and in 99 sides, hemodynamic stenosis (lumen diameter reduction, 50% or greater). Of the 238 carotid artery plaques examined, 167 were homogenous and 71 were heterogenous. During a mean follow-up of 34 months, 27 new focal neurologic deficits occurred. Patients with previous symptoms had a higher incidence of new deficits (18/79 vs 9/135) (p less than 0.01). The severity of the stenosis and the presence of a heterogenous plaque were statistically correlated with the occurrence of new deficits (p less than 0.001). Multivariate analysis showed that the ultrasonographic pattern and the severity of the stenosis were independent variables. We conclude that a heterogenous plaque should be considered an unstable plaque with the possibility of causing cerebral ischemia. PMID- 3051474 TI - Liver transplantation in the treatment of bleeding esophageal varices. AB - From March 1980 to July 1987, 1000 patients with various end-stage liver diseases received orthotopic liver transplants. Of the 1000 patients, three hundred two had definite histories of bleeding from esophageal varices before transplantation. There were 287 patients with nonalcoholic liver diseases and 15 patients with alcoholic cirrhosis. All patients had very poor liver function, which was the main indication for liver transplantation. One- through 5-year actuarial survival rates of the 302 patients were 79%, 74%, 71%, 71%, and 71%, respectively. These survival rates are far better than those obtained with other available modes of treatment for bleeding varices when liver disease is advanced. Long-term sclerotherapy is the treatment of primary choice for bleeding varices. Patients in whom sclerotherapy fails should be considered for liver transplantation unless clear contraindications exist. PMID- 3051475 TI - A randomized, controlled trial to determine the effectiveness of fascial infiltration of bupivacaine in preventing respiratory complications after elective abdominal surgery. AB - A randomized, controlled trial was performed to determine whether infiltration of fascia with bupivacaine, a long-acting local anesthetic, at the time of closure after elective laparotomy, is effective in preventing postoperative respiratory complications. At the Toronto General Hospital 415 patients undergoing elective laparotomy were randomly allocated to receive bupivacaine 0.25% (2 ml/cm incision), infiltrated into the fascia evenly along both sides of the incision before wound closure (202 patients), or to have closure without infiltration (213 patients). Chest x-ray (CXR) films of all patients were obtained preoperatively and on the second postoperative day. Pulmonary function studies were performed preoperatively and for the first two consecutive days postoperatively. CXR films were scored by a blinded observer. Postoperatively, 64% of the treatment group and 56% of the control group had evidence of atelectasis on CXR films (p = NS, chi 2 test). Both groups had similar decrements in vital capacity and expiratory reserve volume on the first and second postoperative days. There was no significant difference in the amount of analgesic taken in the first 24 hours, although the time to first analgesic was significantly longer in the treatment group (2.2 vs 1.3 hours, p less than 0.055). We conclude that infiltration of the fascia with 0.25% bupivacaine at a dose of 2 ml/cm of incision is not effective in preventing postoperative atelectasis. It does not reduce use of an analgesic although it may delay its initial requirement. PMID- 3051476 TI - Creatine kinase as a prognostic indicator in electrical injury. AB - Serial serum creatine kinase (CK) and creatine kinase myocardial band isoenzyme (CK-MB) levels were obtained from 116 of 125 electrical burn patients admitted from 1976 through 1986. We divided patients into three groups (peak CK within 2 days after admission) as follows: group 1, CK less than 400 U/L; group 2, CK = 400 to 2500 U/L; group 3, CK greater than 2500 U/L. Clinical myocardial infarction (MI) was determined by ischemic ECG changes, LDH isoenzyme patterns, and clinical course. Skin grafts occurred in 2 of 24 patients from group 1, in 15 of 31 from group 2, and in 37 of 61 from group 3. Hospital stay (mean +/- SEM) was 4.6 +/- 1.3 days for group 1, 20.2 +/- 5.4 for group 2, and 37.7 +/- 3.6 for group 3. Group 1 patients required no amputations; group 2 had 1 limb and 5 digit amputations; group 3 had 22 limb and 16 digit amputations. Only three clinical MIs were found (all in group 3), although 1 of 31 patients from group 2 and 32 of 61 from group 3 had CK-MB greater than 4%. Highly elevated CK and CK-MB are associated with longer hospitalization, and a greater risk of skin grafting or amputation, than with levels less than 400 U/L. Clinical MI is rare and cannot be diagnosed by elevated CK-MB alone. PMID- 3051477 TI - Gastrointestinal complications after cardiac surgery. AB - Gastrointestinal (GI) complications after cardiac surgical procedures are infrequent but severe. Thirty-three GI complications were identified in 25 patients who underwent cardiac surgery during a 7-year period (2.0% incidence). The mortality rate for patients having these GI complications was 44%. Acute acalculous cholecystitis was the most lethal complication (86%). Acute pancreatitis was the most common complication (eight patients). Most patients responded well to conservative measures. Five patients had upper GI hemorrhage and three had lower GI bleeding that required more than 2 U of packed red blood cells. All patient conditions were diagnosed endoscopically and none necessitated operation. Of the remaining patients, one was operated on because of perforated duodenal ulcer, one because of perforated diverticulitis, and one because of pseudo-obstruction of the colon, and one patient underwent diagnostic laparotomy and showed negative results for presumed acalculous cholecystitis. Liver failure was fatal in all three patients in whom it occurred. GI complications correlated significantly with advanced age, prolonged bypass times, valve surgery, and the female sex. We conclude that septic GI complications--particularly acute acalculous cholecystitis and perforated viscus--after cardiac surgery are uncommon but lethal. Clinical features are often subtle, and a high index of suspicion is necessary for an early diagnosis and the institution of appropriate treatment. PMID- 3051478 TI - Gastric bypass revision: lessons learned from 920 cases. AB - Roux-en-Y gastric bypass (RGB) is an accepted operation for the control of body weight in morbidly obese patients. Early technical complications and inadequate weight loss, well-known sequelae of this procedure, necessitated reoperation in 42 patients of 920 who underwent RGB in a 10-year period. Indications for reoperation included dilated gastrojejunal anastomosis (16), inadequate weight loss without demonstrable enlargement of the anastomosis (10), staple line breakdown (6), anastomotic obstruction (4), anastomotic leak (4), and enlarged proximal gastric pouch (2). Reoperation consisted of completely redoing the initial RGB in 20 patients, redoing the anastomosis alone in 17 patients, staple line revision in four patients, and intraoperative dilatation of the anastomosis in one patient. After initial RGB, 26 of the 42 patients (61.9%) experienced major complications. After revision of RGB, there were major complications in 21 patients (50%). In conclusion, major postoperative complications may contribute to RGB failure, RGB revision for early technical failure or inadequate weight loss is associated with a high incidence of major complications and, subsequently, negligible weight loss. Therefore repair of RGB for technical failure or complications is not recommended. PMID- 3051479 TI - [Changing the work environment needs involvement, economics and planning]. PMID- 3051480 TI - [Occupational diseases are an ancient concept]. PMID- 3051481 TI - [Transplantation. New bone marrow, a difficult process for everyone]. PMID- 3051482 TI - [Danish Nursing Council's nursing education courses]. PMID- 3051483 TI - [In memoriam I. A. Kassirskii, Academician of the Academy of Medical Sciences of the USSR (on the 90th anniversary of his birth)]. PMID- 3051484 TI - [Pharmacological characteristics of cyclosporin A. Its use in kidney transplantation]. PMID- 3051485 TI - [Endogenous regulators of hematopoiesis and their role in the therapy of leukemias]. PMID- 3051486 TI - [Effect of interleukin 2 on blast proliferative activity and expression of T-cell markers in patients with acute lymphoblastic leukemia]. AB - The proliferative activity of blast cells from patients with acute lymphoblastic leukemia was compared with the activity of lymphoid cells of the thymus and blood of healthy subjects. The spontaneous proliferative activity in cells obtained extempore varied widely. On the basis of this index groups of patients with high proliferative activity (group 1) exceeding the level of thymocyte proliferation and with low proliferative activity (group 2) were distinguished. Direct correlation was revealed between the initial spontaneous proliferative activity and the number of leukocytes as well as with the absolute number of blasts in the blood. A decrease in the initial high proliferative activity in the time course of cultivation was noted in some patients of group 1 (group 1B). In the majority of these patients (91%) the administration of interleukin-2 into the cultural medium prevented a decrease in the proliferative activity. In the other patients of group 1 (group 1A) a high spontaneous proliferative activity was not decreased during cultivation. In group 1A a proliferative response to T-cell growth factor was observed in 36% of patients. The patients' blast cells in group 2 did not respond to this lymphokine. The expression de novo of T-cell markers was observed in some patients of group 1 as a result of interleukin-2 action. The role of interleukin-2 in the pathogenesis of acute lymphoblastic leukemia is discussed. PMID- 3051487 TI - [Transplantation of embryonic liver in severe combined immune deficiency]. PMID- 3051488 TI - [Interleukin 2]. PMID- 3051489 TI - [The role of the opsonophagocytic system in the pathogenesis of sepsis]. PMID- 3051490 TI - [General principles of research on the hemostatic system and analysis of new methods for detecting intravascular blood coagulation]. PMID- 3051491 TI - Split notochord syndrome with dorsal enteric fistula and sacral agenesis. AB - Split notochord syndrome of the lumbosacral spine in association with dorsal enteric fistula is a rare phenomenon. To date, only nine human cases have been reported in the English literature. We present another case of this type, with sacral agenesis as an additional and unique finding. Several etiological theories are discussed including the persistence of the neurenteric canal, the occurrence of an ectopic or accessory neurenteric canal, a division or local redundancy of the notochord, an entodermal-ectodermal adhesion, neural tube rupture caused by oversecretion of fluid, and failure or aberrancy of dorsal aortic distribution to the region of the neural folds resulting in prevention of timely neural tube closure. PMID- 3051492 TI - Postnatal uterine development in the rat: estrogen and antiestrogen effects on luminal epithelium. AB - We examined the effects of the synthetic estrogens, diethylstilbestrol (DES) and ethynylestradiol (EE), and the triphenylethylene antiestrogen, clomiphene citrate (CC), on uterine growth and development in the rat. These compounds, unlike estradiol, do not bind significantly to rat serum alphafetoprotein (AFP). Administration of DES or EE during the period of normal uterine gland genesis (postnatal days 10-14) induced luminal epithelium hypertrophy and increased uterine wet weight. The durations of these responses were dose-related. By day 26, luminal epithelium cell numbers were significantly depressed, compared to controls. Uterine gland development was delayed 6 to 9 days, depending upon estrogen dose, and the numbers of uterine glands ultimately achieved were generally less than in untreated control animals. While a daily dose of 0.1 micrograms CC/rat did not alter uterine development, 10 micrograms CC/rat caused prolonged luminal epithelium hypertrophy and inhibited uterine gland genesis without inducing the large increases in uterine weight or the decreases in luminal epithelium cell number seen after estrogen exposure. The number of stromal cells was significantly increased on day 26 after CC exposure. Together with previous studies, these data demonstrate the greater potency and developmental stage specificity of non-AFP-bound estrogens with respect to altering uterine gland development. In addition, these data suggest that the disruptive influence of antiestrogens on gland genesis may be mediated through an indirect influence on the uterine stroma. PMID- 3051493 TI - Teratoma of the fallopian tube presenting as a free-floating pelvic mass. PMID- 3051494 TI - Psychiatric disorders in HIV-spectrum illness. PMID- 3051495 TI - Historic hospital a monument to humane mental health care. PMID- 3051496 TI - Hospitals and HCFA--a terminal situation? PMID- 3051497 TI - [The use of physical agents in the treatment of spasticity]. PMID- 3051498 TI - [The value of retraining in female urinary incontinence]. PMID- 3051500 TI - [Indications for routine examinations in the physician's office]. PMID- 3051499 TI - [General principles in interpreting diagnostic tests]. PMID- 3051501 TI - [Hematology in general practice]. PMID- 3051502 TI - [Urinalysis in the physician's office: methods and interpretation]. PMID- 3051503 TI - [Ultrasound diagnosis: principles, possibilities and limits]. PMID- 3051504 TI - [Assessing infections in general practice]. PMID- 3051505 TI - [Cardiologic assessment in general practice]. PMID- 3051506 TI - [Diagnostic methods in gastroenterology in the physician's office]. PMID- 3051507 TI - [Evaluating lung function in medical practice]. PMID- 3051508 TI - Influence of erdosteine, a mucolytic agent, on amoxycillin penetration into sputum in patients with an infective exacerbation of chronic bronchitis. AB - Twenty four patients with acute infective exacerbations of chronic bronchitis received amoxycillin alone or in combination with erdosteine (a mucolytic agent) for a week in a double blind, placebo controlled study. Clinical assessment scores, body temperature, serum and sputum amoxycillin concentrations, and sputum culture results were recorded in each group. Erdosteine significantly increased antibiotic concentrations in sputum but not in serum. The combined treatment also caused a more rapid decrease in sputum viscosity and in body temperature and faster sterilisation of the sputum. These results show that erdosteine increases amoxycillin concentration in sputum in patients with acute exacerbations of chronic bronchitis. This effect may be due to a reduction in the viscosity of the bronchial secretions produced by erdosteine. PMID- 3051509 TI - Comparison of histamine and methacholine for use in bronchial challenge tests in community studies. AB - Measurement of bronchial reactivity is widely used in epidemiological surveys. Histamine has been compared with methacholine inhalation challenge in two samples of adults from a small town to determine which is the better agent for use in community studies. Increasing doses of histamine and methacholine were given, up to a maximum of 4 and 12 mumol respectively, according to the method of Yan et al, the provocative dose of agonist causing a 20% fall in FEV1 (PD20) being measured. More subjects had a measurable PD20 with methacholine than with histamine, both in a random sample of 108 subjects (25 v 11 subjects, p less than 0.01) and in an additional 95 subjects selected because of wheeze in the last 12 months (67 v 48 subjects, p less than 0.01). Side effects were mild with both agents but histamine caused voice change in more subjects (21% v 11%). Repeatability was assessed in a further group of subjects with wheeze in the last year. The 95% range for a single estimation of PD20 in subjects with a measured PD20 on at least one occasion was +/- 2.5 doubling doses for histamine (n = 25) and +/- 2.1 doubling doses for methacholine (n = 33). Thus methacholine has advantages over histamine for community studies of bronchial reactivity as it is possible to use doses that produce more PD20 measurements with fewer side effects. PMID- 3051510 TI - Abolition of methacholine induced bronchoconstriction by the hyperventilation of exercise or volition. AB - Total pulmonary resistance was measured from continuous records of flow and oesophageal pressure in five normal subjects on three separate days before and after inhalation of methacholine. The dose of methacholine produced, on average, a fivefold increase in airway resistance. Immediately after methacholine inhalation the subjects underwent a progressive exercise test on a cycle ergometer (day 1) or voluntary hyperventilation (day 2) or remained resting (day 3). On the first day during exercise pulmonary resistance fell rapidly to baseline levels within two to three minutes and remained there for the 10 minute duration of the exercise. On day 2 voluntary reproduction of the same level and pattern of ventilation as during exercise resulted in a similar fall of resistance. On the third day, when the subjects remained at rest, pulmonary resistance remained raised for 10 minutes. It is concluded that the bronchodilator effects of exercise can be explained by the increased ventilation rather than the exercise itself, but with much smaller tidal volumes than have previously been thought necessary to reduce drug induced bronchoconstriction. PMID- 3051513 TI - [Chancre: an old-fashioned concept or a modern solution for sleeping sickness?]. AB - A brief introduction on trypanosomiasis in ruminants and an explanation of the term chancre are followed by a discussion of a number of findings on the early pathogenesis of the disease. This study formed part of the requirements in obtaining a doctorate in veterinary medicine. PMID- 3051512 TI - In memoriam Zbigniew Stanislaw Latallo 1924-1987. PMID- 3051511 TI - Pulmonary Kaposi's sarcoma in two recipients of renal transplants. AB - Among 350 recipients of renal transplants seen at the Riyadh Military Hospital, 12 developed Kaposi's sarcoma. Two of these sarcomas presented primarily as a problem of diffuse lung infiltrates in an immunocompromised host. In one the diagnosis was established by transbronchial lung biopsy. Withdrawal of immunosuppression led to satisfactory radiological resolution in both patients. PMID- 3051514 TI - [Fraud in medicine with the child as victim. Munchausen syndrome by proxy. A review of the literature]. AB - In a review of the literature on the subject 'Munchhausen syndrome by proxy' the authors try to give an impression of the history of diagnosis and treatment of this special kind of child abuse. Particularly aspects of confrontation are discussed. They consider features of the chief actors in this very dramatic play. The authors point to problems in differential diagnosis in the framework of other subjects like 'non-accidental poisoning', 'doctor-shopping' and 'filicide'. They discuss ethical and legal consequences. PMID- 3051515 TI - [Childhood diseases and pediatrics in the notebooks of Constantijn Huygens (1595 1687)]. AB - In this article we collected and commented on the passages referring to children's diseases and paediatrics in the notebooks of the well-known Dutch statesman and poet Constantijn Huygens. This remarkable source for the history of paediatrics describes the psychological and physical development of Huygen's five children and the diseases with which they were afflicted. PMID- 3051517 TI - Chlorinated drinking water is mutagenic and causes 3-methylcholanthrene type induction of hepatic monooxygenase. AB - Acid/neutral fractions of 4 chlorinated drinking water samples were tested for mutagenicity in the Ames' test and injected intraperitoneally to 10- and 20-day old Wistar rats at doses of 200 and 100 liters of water/kg body weight. Cytochrome P-450 mediated enzyme activities of ethylmorphine-N-demethylase (EMND), 7-ethoxycoumarin-O-deethylase (ECOD), 7-ethoxyresorufin-O-deethylase (EROD) and 7-pentoxyresorufin-O-dealkylase (PEROD) were determined in the 9000 g supernatant fraction of liver homogenate. EROD was introduced by the concentrates. The induction was related to the mutagenic activity. About 4-fold increase in activity was observed with the most mutagenic sample. PEROD was also slightly enhanced. EMND and ECOD activities were not affected by the lower dose, but the higher dose caused inhibition of 30-40%. Although the extracts were not toxic to bacteria, they were unexpectedly toxic to rats. It is concluded that the samples contained 3-methylcholanthrene (3-MC) type inducer(s). PMID- 3051516 TI - Effects of acute administration of O,S,S-trimethyl phosphorodithioate on the generation of cellular and humoral immune responses following in vitro stimulation. AB - The time course of immune modulation induced by acute treatment with O,S,S trimethyl phosphorodithioate (OSS-TMP), an impurity in technical formulations of malathion, was examined in female C57BL/6 mice. The immune parameters studied included the generation of cytotoxic T lymphocytes (CTL) to alloantigen (H-2 incompatible) and antibody secreting cells to sheep red blood cells, proliferative response to the mitogens, and interleukin-2 (IL-2) production. Acute administration of the non-toxic doses of OSS-TMP, i.e. 20 or 40 mg/kg, led to an elevation in the generation of a CTL response on day 1 or 7, respectively. At 20 mg/kg OSS-TMP, the antibody response was elevated at day 3. However, at a dose of 40 mg/kg OSS-TMP, the antibody response was suppressed at day 1 following treatment. Following acute administration of 60 or 80 mg/kg OSS-TMP, the generation of an antibody and CTL responses was suppressed at all time points tested with 1 exception. One day following treatment at a dose of 60 mg/kg OSS TMP, there was no change in the CTL response. At day 7 following treatment, the mitogenic responses to lipopolysaccharide and phytohemagglutinin were elevated at all doses of OSS-TMP administered. At this time point, however, the proliferative response to Concanavalin A was elevated in a dose dependent manner. IL-2 production was suppressed following acute administration of 60 or 80 mg/kg OSS TMP at all time points tested and at all doses tested on day 5 following treatment. These data indicate that OSS-TMP, unlike its congener, O,O,S-trimethyl phosphorothioate, enhances the generation of humoral and cell mediated immune responses of C57BL/6 mice following administration of non-toxic doses. PMID- 3051518 TI - Physiological pharmacokinetic models: some aspects of theory, practice and potential. AB - Models are intellectual constructs that pattern selected relationships among the elements of one system to correspond in some way to elements of a second system. In pharmacokinetics, physiological models provide a clearly articulated, rational, explanatory basis for the integration of empirical data; they do this by partitioning the biological system into relevant components (tissues, organs, etc.) and linking them together through the circulatory system. Unlike conventional mammillary compartment models, there is a clear correspondence between model system elements and physiological entities. By virtue of their high degree of physical and biochemical relevance, these models can help provide deep insight into structure, function and mechanism. Pharmacokinetic (and potentially pharmacodynamic) response-time relationships can thus be understood in terms of interconnections and behavior of constituent subsystems. At their worst, these models provide stale or infertile views of reality and thus frustrate and alienate us with the triviality of their insights. At their best, they allow us to understand the accumulation of thought in pharmacokinetics and pharmacodynamics, and help with the integration of data and improvement of experimental design. PMID- 3051519 TI - Respiratory dose-response study of normal and asthmatic volunteers exposed to sulfuric acid aerosol in the sub-micrometer size range. AB - Twenty-one healthy and 21 asthmatic volunteers were exposed to respirable sulfuric acid aerosol (mass median particle diameter approximately 0.9 micron, geometric standard deviation 2.5) in a chamber at 21 degrees and 50% relative humidity. Measured sulfuric acid concentrations averaged 0, 380, 1060, and 1520 micrograms/m3 (in the occupational range, higher than concentrations observed in ambient air pollution). Exposures to different concentrations occurred in randomized order 1 week apart. They lasted 1 hr and included three 10-min periods of heavy exercise. Healthy volunteers showed no statistically significant changes in pulmonary function, airway reactivity to inhaled methacholine, or overall reporting of irritant symptoms which could be attributed to acid exposure. They did show a slight statistically significant (P less than .01) increase in cough with increasing acid concentration. At the two highest acid concentrations, asthmatics showed significant increases in irritant symptoms and decrements in pulmonary function, without significant changes in airway reactivity. Their function decrements appeared to increase with time during exposure. Previous studies in fog (10 degrees, median particle diameter approximately 10 micron) with similar concentrations of sulfuric acid showed more symptoms but less pulmonary function change, perhaps reflecting different sites of particle deposition in airways and/or different degrees of neutralization by airway ammonia. This and earlier evidence predicts little, if any, acute irritant response in short-term (1 hr or less) exposures to sulfuric acid at concentrations found in ambient air pollution. PMID- 3051520 TI - Review of pesticides: chemistry, uses and toxicology. PMID- 3051521 TI - Cisplatin pharmacokinetics: applications of a physiological model. AB - A physiological pharmacokinetic model for the disposition of the antineoplastic drug cis-diamminedichloroplatinum(II) (cisplatin or DDP) in several mammalian species is reviewed. The significance of the model's key parameters and of their interspecies relationships is discussed. Methods for estimating two of these parameters (the rate constants for formation of the fixed and mobile metabolites) from in vitro experiments are presented. Fixed and mobile metabolites are formed by the irreversible binding of cisplatin to macromolecules and low molecular weight nucleophiles, respectively. Use of the model to simulate and predict the pharmacokinetic behavior of cisplatin and its metabolites in different animal species following intravenous and intraperitoneal administration is illustrated. Fixed metabolite formation rate constants can be used in conjunction with mass transport parameters to estimate tissue exposures to cisplatin following systemic or regional drug administration. The model logically can serve as the basis for a pharmacodynamic model that incorporates cisplatin reactions with DNA. The model also provides a means for comparing the pharmacokinetic characteristics of cisplatin analogs and for assisting in rational analog development. PMID- 3051522 TI - Cell growth dynamics in long-term bladder carcinogenesis. AB - A biologically based probabilistic model of the carcinogenic process has been developed based on a two-stage theory of carcinogenesis. The model has been validated utilizing experimental urinary bladder carcinogenesis studies in the rat, with an emphasis on quantification of cell dynamics. Critical parameters tracked through this process include mitotic rates, cell loss and birth rates, and irreversible cellular transitions from normal to initiated to transformed states. Analyses demonstrate the sensitivity of tumor incidence to the timing and magnitude of changes to these cellular variables. Modeling has been applied to genotoxic compounds, such as N-[4-(5-nitro-2-furyl)-2-thiazolyl]formamide, and non-genotoxic compounds, such as sodium saccharin. For the latter compounds, complex administration regimens have been studied, including two-generation experiments, initiation-promotion experiments, and sodium saccharin administration following ulceration and regenerative hyperplasia. Modeling indicates that the effects of such compounds can be explained entirely on the basis of cytotoxicity and consequent hyperplasia. Quantitative modeling based on biological processes has the potential for direct application to carcinogenic risk assessment. PMID- 3051523 TI - Computational approaches to the identification of suspect toxic molecules. AB - We have presented computational approaches that can be used for the relatively rapid identification of suspect toxigens, including carcinogens, in two different classes of compounds: (a) halogenated olefins and epoxides, and (b) substituted dibenzo-p-dioxins. A common element in these approaches is the key role played by the molecular electrostatic potential. It is applied in two different ways, however; it is used to assess the reactivity of a specific site in the case of the epoxides, and for the dibenzo-p-dioxins the focus is on the overall pattern of negative regions above the molecular plane. While we are continuing to develop and refine both types of analysis, especially that related to the dibenzo-p dioxins, the results obtained so far are encouraging, and indicate that these can be regarded as useful screening techniques for identifying compounds that require further and more exhaustive investigation. PMID- 3051524 TI - Quantitative sensory assessment in toxicology and occupational medicine: applications, theory, and critical appraisal. AB - Sensory systems are affected by a number of chemicals. Their functions can be quantitatively assessed with psychophysical methods. Psychophysics is the scientific study of the relationship between the physical dimension(s) of a stimulus and the behavioral response it generates. First, the unique contribution of psychophysics to toxicology is illustrated with a few examples taken from the fields of audition, vision, and somatosensory sensitivity. Then, a brief survey of the evolution of psychophysics is presented and followed by a review of its basic scientific foundations. Psychophysical evaluation is accomplished by delivering quantified stimuli in a prespecified order and by requesting a standardized response from the subject. Psychophysical methods specify the sequence of presentation of stimuli, and include the method of limits, the method of constant stimuli, as well as the adjustment and tracking methods. The response paradigms specify the standardized response the subject has to make to stimuli presented according to a specific format. The three major response paradigms are the yes-no, forced-choice, and rating paradigms. Sensory processes can be studied in humans as well as in animals by several techniques, such as classical conditioning, operant behavior, and reflex modulation. A critical review of the application of psychophysical methods and response paradigms is developed. Several examples illustrate the potential confusion brought about by unorthodox use of psychophysical terminology and methodology. Finally, a few suggestions are given for the evaluation of sensory processes in humans. PMID- 3051525 TI - Quantitation of naturalistic behaviors. AB - Naturalistic behaviors are behaviors that organisms exhibit 'in nature'. Eating, sleeping and sexual behaviors are examples. Since naturalistic behaviors are observed to occur without any apparent training or learning, some people mistakenly believe that all naturalistic behaviors are unlearned, and are thus different from laboratory behaviors. We maintain that naturalistic behaviors can be studied profitably in the toxicological laboratory, using quantitative techniques from behavioral neuroscience. Understanding of toxicity and underlying mechanisms is enhanced when naturalistic behaviors are thought of as responses to stimuli. Stimuli that influence naturalistic behaviors may arise inside the organisms (e.g., physiological signals of hunger) or outside the organisms (e.g., the smell of food or the start of the nocturnal lighting cycle). A practical, noninvasive, automated system can be used to improve upon the cage-side observation currently used to evaluate naturalistic behaviors in toxicity screening. Effects of alkyltins and other neurotoxicants upon eating, drinking, rearing, and the daily cycle of rest-activity will be shown. The rodent's pattern of nocturnal activity has proven to be particularly sensitive to neurotoxicants, and thus deserves additional attention in developing neurobehavioral toxicology. PMID- 3051526 TI - Quantification of operant behavior. AB - The study of performance on intermittent schedules of reinforcement has proved to be a powerful tool in the fields of experimental psychology and behavioral pharmacology and presently is proving equally valuable in behavioral toxicology. The ability to specify precisely contingencies of reinforcement allows a careful and detailed quantification of performance. Intermittent schedules of reinforcement may be used in behavioral toxicology in a number of ways. A baseline of performance may be established and utilized to monitor acute effects or to track effects of chronic exposure to a toxic agent. Alternatively, schedules of reinforcement may be used in experiments requiring group comparisons, where both terminal performance and acquisition of performance may be of interest. The use of different schedules, generating different rates and patterns of performance, may be compared to elucidate behavioral mechanisms. Use of computers for schedule control and data acquisition allows a detailed analysis of performance, thus increasing the probability of the detection of subtle effects. PMID- 3051527 TI - Aspects of biological monitoring of exposure to glycol ethers. AB - Glycol ethers are frequently used as solvents, detergents, and emulsifiers alone or as components in industrial and consumer products. The monomethyl and monoethyl ethers of ethylene glycol, and their acetate esters, are teratogenic and embryotoxic and cause testicular damage in laboratory animals, while the monobutyl ether causes hemolysis of the red blood cells. The adverse effects are attributed to the acid metabolites methoxy-, ethoxy- and butoxyacetic acid, respectively. The glycol ethers may readily enter the body by inhalation as well as dermal uptake. Biological monitoring of exposure to glycol ethers has therefore been suggested. This paper reviews physical properties, occurrence, analysis, toxicity, and toxicokinetics of the most common glycol ethers and then discusses toxicokinetic aspects of biological monitoring. The effect of physical exercise and the relative importance of respiratory and percutaneous absorption on the internal exposure to glycol ethers are illustrated. Monitoring the acid metabolite in urine is suggested as the best index of exposure. Intra- and inter individual variability, dose-dependent toxicokinetics, and metabolic induction and inhibition are examples of possible sources of error in the estimation of internal exposure from the urinary excretion of acid metabolite. PMID- 3051528 TI - A review of propylene glycol dinitrate toxicology and epidemiology. AB - Propylene glycol dinitrate (PGDN) is a rapidly metabolized, nitrated ester explosive propellant with acute cardiovascular effects at lower levels of exposure and methemoglobinemia and vascular collapse at higher ones. Exposure can be by dermal or inhalation routes. Toxicology has played an important role in setting workplace permissible exposure limits, which have been limited by vascular headaches and subtle, transient decrements in central nervous system performance. Less well characterized is the etiology of excess, long-term cardiac morbidity in PGDN-exposed workers. Human central nervous system degeneration and links to infectious diseases are unsubstantiated concerns. PMID- 3051529 TI - The Canadian American Ticlopidine Study (CATS) in thromboembolic stroke. Design, organization, and baseline results. AB - The Canadian American Ticlopidine Study is a randomized, placebo-controlled, double-blind, multicenter study to assess the efficacy and safety of ticlopidine hydrochloride in patients who have suffered a thromboembolic stroke no less than 1 week and no more than 4 months before entry into the study. The primary assessment of efficacy will be based on the cluster of outcome events recurrent stroke, myocardial infarction, or vascular death. Twenty-five clinical centers, 12 in Canada and 13 in the United States, entered a total of 1,072 patients during a 3-year recruitment period; these patients were randomly allocated to receive either 250 mg ticlopidine or identical-appearing placebo tablets twice daily for up to 3 years. Patient recruitment was completed in December 1986. Patients were followed for a maximum of 3 years or until the close of the study in December 1987; at that time an average follow-up of 25 months had been achieved. We summarize the protocol and organization of the study and document the methods of execution and analysis, with corresponding criteria, before disclosure of the treatment code to any of the study investigators. We also provide a clinical description of the patients at entry into the study. PMID- 3051530 TI - Danish very-low-dose aspirin after carotid endarterectomy trial. AB - The effect of very-low-dose aspirin as an antithrombotic agent was evaluated blindly in 301 patients who had recently undergone carotid endarterectomy. After randomization, 150 patients received aspirin and 151 received placebo. The two groups were comparable with regard to age, sex, blood pressure, previous cerebrovascular events, and smoking habits. The effect of the study medication on platelet aggregation was measured twice in each patient during the first 2 months and at each follow-up visit; the dose was individually adjusted. In 76% of the patients receiving aspirin, 50 mg/day gave satisfactory platelet inhibition, 13% needed 60 mg/day, 8% needed 70 mg/day, and 3% needed 100 mg/day. Platelet aggregation was found to be inhibited in only 1.2% of the measurements in the patients receiving placebo. Observation during treatment averaged 21 months; total intention-to-treat follow-up averaged 25 months. For the combined outcome events of transient ischemic attack, stroke, acute myocardial infarction, and vascular death, aspirin reduced risk by 11% (95% confidence limits: -38% to 48%, p greater than 0.1). Thus, there was no significant effect of very-low-dose aspirin in our trial. PMID- 3051531 TI - Ocular bruits in ischemic cerebrovascular disease. AB - A total of 72 ocular bruits in 50 patients with symptoms of atherothrombotic ischemic cerebrovascular disease were studied with continuous-wave Doppler ultrasonography with spectrum analysis (Dopscan). Fourteen patients also had conventional angiography, and 14 had digital subtraction angiography. Ocular bruits by augmentation flow due to occlusion (seven bruits, 9.7%) or tight stenosis (17 bruits, 23.6%) of the contralateral internal carotid artery accounted for only 24 ocular bruits (33.3%). In contrast, siphon stenosis ipsilateral to the ocular bruit was a very common finding. All 14 patients studied with conventional angiography had variable degrees of siphon stenosis ipsilateral to the ocular bruits; there was one angiography failure. We conclude that siphon stenosis can cause ocular bruit alone or can act in combination with augmentation flow by contralateral carotid occlusion or tight stenosis. The difference in their relative occurrence in our patients compared with previous reports probably reflects racial differences in the distribution of stenotic or occluded lesions of the carotid artery between our patients and Caucasian patients. The ocular bruit was the only auscultatory finding in more than a quarter of our patients (14 of 50, 28%). PMID- 3051532 TI - Calcium channel blockers correct acidosis in ischemic rat brain without altering cerebral blood flow. AB - We compared the effects of intravenous infusions of 40 micrograms/kg/min verapamil (n = 5), 0.5 microgram/kg/min nimodipine (n = 5), and 5 ng/kg/min prostacyclin (n = 6) and no treatment (n = 6) on local cerebral pH and local cerebral blood flow in middle cerebral artery-occluded rats 90 minutes after the ischemic insult. Local cerebral pH and local cerebral blood flow were determined simultaneously by a double-label autoradiographic technique. The infusions were started 15 minutes after completion of the occlusion and ended at decapitation 90 minutes after completion of the occlusion. Cortical pH for four regions in the ischemic middle cerebral artery territory of rats receiving verapamil or nimodipine was normalized (mean +/- SEM 6.90 +/- 0.02 and 7.01 +/- 0.01, respectively, for the parietal, sensorimotor, frontal, and auditory cortexes), while mean +/- SEM pH in rats receiving prostacyclin was 6.79 +/- 0.01; in untreated rats, mean +/- SEM pH in the same brain regions was 6.72 +/- 0.01. Local cerebral pH in the verapamil- or nimodipine-treated rats was thus significantly different from that in untreated rats (p less than 0.05). Local cerebral blood flow in treated rats was not different from that in untreated ones. Our findings suggest that calcium channel blockers correct ischemic cerebral acidosis by metabolic mechanisms rather than by changes in blood flow. PMID- 3051534 TI - White matter signal abnormalities in normal individuals: correlation with carotid ultrasonography, cerebral blood flow measurements, and cerebrovascular risk factors. AB - We studied 52 asymptomatic subjects using magnetic resonance imaging, and we compared age-matched groups (51-70 years old) with and without white matter lesions with respect to carotid ultrasonography, cerebral blood flow (xenon-133 injection), and cerebrovascular risk factors. In the group with white matter signal abnormalities, we noted a higher frequency of extracranial carotid artery disease, a lower mean gray matter blood flow (F1), and a significant reduction (p less than 0.05) in blood flow of the slow-flowing (F2) compartment. Hypertension, diabetes mellitus, and cardiac diseases (p less than 0.002) were found more often in this group. Our results indicate that a higher incidence of changes known to be associated with an increased risk for stroke exists in the presence of white matter lesions in normal elderly individuals. PMID- 3051533 TI - Stable prostacyclin analogue preventing microcirculatory derangement in experimental cerebral ischemia in cats. AB - We evaluated the effect of a stable synthetic prostacyclin analogue, TRK-100, on the microcirculatory derangement occurring in feline pial vessels with endothelial damage after middle cerebral artery occlusion. Fifteen adult cats were divided into an untreated group (Group 1, n = 8) and a treated group (Group 2, n = 7). Thirty minutes after 10 minutes of ultraviolet irradiation, which selectively damaged endothelium in the pial vessels, the middle cerebral artery was occluded in both groups and maintained for 30 minutes. In Group 2, 50 ng/kg/min TRK-100 was continuously infused intravenously following ultraviolet irradiation. In both the pial arteries and veins, platelet aggregate adhesion to the endothelium with subsequent thrombus formation was significantly (p less than 0.01 and p less than 0.05, respectively) inhibited during middle cerebral artery occlusion in Group 2 compared with Group 1. Similarly, blood flow stasis in the pial veins was effectively prevented in Group 2 during occlusion. Furthermore, the pial artery diameter returned to the control level during the late period of occlusion, whereas in Group 1 the pial artery remained constricted. Our data suggest that TRK-100 can prevent microcirculatory derangement in the acute stage of ischemic stroke. PMID- 3051535 TI - Dementia due to vascular disease--a multifactorial disorder. AB - This review was undertaken to evaluate critically the literature pertaining to vascular dementia with the objective of determining a more useful and scientifically supported definition of vascular dementia, its relation to other causes of dementia, and the biologic mechanisms involved in its causation. PMID- 3051537 TI - Evaluation of tricyclic antidepressant plasma levels by an automated enzyme immunoassay (EMIT) in comparison to a high-performance liquid chromatographic method. AB - A new homogeneous enzyme immunoassay technique (EMIT) for the measurement of plasma levels of amitriptyline, nortriptyline, imipramine, and desipramine was used with an automated procedure and the results were compared to those of a high performance liquid chromatographic (HPLC) method. Precision of the EMIT test was similar to that of the HPLC method with within-day coefficients of variation in the range of 3.9-10.9% (EMIT) and 3.9-9.6% (HPLC). The day-to-day coefficients of variation ranged from 4.4 to 11.7% for EMIT and from 6.1 to 8.4% for HPLC. Samples from 124 patients were analyzed by both methods and a good correlation was observed for all the four drugs. A paired t test indicated no significant difference for the EMIT and HPLC values. No significant interferences were observed between the tricyclics tested and other commonly associated drugs, such as benzodiazepines and neuroleptics. The new EMIT assay proved to be rapid and easy to perform and showed sufficient reliability and reproducibility to be used for either emergency or routine analysis. PMID- 3051536 TI - Women's health: an alternative perspective for choosing interventions. AB - This paper outlines the health problems of mothers, discusses the links between maternal health and child health, and emphasizes the need to focus attention more clearly on the problems of women and the interventions that might help them as a way to improve both maternal and child health. The special problems of girls and women in the developing world--including maternity care, abortion, and maternal mortality and morbidity--and the ways in which these problems affect mothers and their children, are examined. Nutritional morbidity and infectious morbidity are described in terms of their effects on maternal and infant health, including low birth weight. It is shown how the cultural, social, and economic factors that affect women and children interact with their health problems. Recommendations are made to: reexamine well established interventions to determine if new program designs might improve long-term results; conduct research on the efficacy of retraining health personnel; broaden and improve service delivery; reassess the cost-effectiveness of highly targeted interventions; reexamine the locus of certain interventions; and emphasize long-term as well as short-term results. Attention should be given to women's health not only during pregnancy, but throughout the life cycle. PMID- 3051538 TI - No effect of influenza vaccination on theophylline pharmacokinetics as studied by ultraviolet spectrophotometry, HPLC, and EMIT assay methods. AB - The effect of influenza vaccination on steady-state pharmacokinetics of theophylline was studied in six healthy young adults by comparing pharmacokinetic parameters found on days 4 and 5 during a 5-day course of theophylline alone with those obtained after influenza vaccination on day 4 of a second study phase. Theophylline plasma concentrations were measured by means of high-pressure liquid chromatography (HPLC) analysis and, in part, with a manual ultraviolet spectrophotometric method and with an EMIT assay. On the fourth and fifth days of each of the two periods of drug administration, theophylline plasma concentration time curves were evaluated, and the following pharmacokinetic parameters were compared: trough plasma concentration (cmin), peak plasma concentration (cmax), time to peak (tmax), and the area under the curve during a dosing interval (AUC). None of these pharmacokinetic parameters of theophylline before and after vaccination were found to be significantly different with any of the analytical methods. PMID- 3051539 TI - Do circulating OKT6-reactive cells belong to Langerhans cell lineage? AB - A few studies have been made in characterizing the phenotype, ontogeny, ultrastructure, and cytochemistry of circulating cord blood cells. Such studies have been hampered by the difficulty in obtaining pure populations of these cells. We therefore sought to obtain T6-positive cell-enriched populations from human cord blood using panning method. In cord blood, dendritic OKT6-labeled cells were classified in two types: some were similar to lymphocyte-like cells and some were consistent with monocyte-like cells. Both types lacked intracytoplasmic Birbeck granules. Homogeneous labeling of HLA-DR antigens were observed in both types. Heterogeneous labeling of T6 antigen was observed in both types. The monocyte-like cells have developed Birbek granule-like structures after exposure to OKT6 and immuno-adherence. The results suggest several different putative lineages of these OKT6-reactive cells: early precursors of dendritic cells, immature neonatal lymphoid subsets, and immature macrophagic leukocytes. PMID- 3051540 TI - Tropical sprue in southern India. AB - Tropical sprue, a primary malabsorption syndrome affecting residents and visitors to several tropical regions, occurs in southern India in endemic and epidemic forms. The stomach, the small intestine and colon are affected and malabsorption results in nutrient deficiency. Enterocyte damage, the primary lesion in southern Indian tropical sprue, is the result of a persistent lesion of the stem cell compartment. This lesion occurs on a background of tropical enteropathy and the available evidence suggests that an immunity conferring agent may be responsible for initiating the damage. PMID- 3051541 TI - A quantitative analysis of the diagnostic value of diethylcarbamazine provocation in endemic Wuchereria bancrofti infection. AB - The efficiency (delta) of the day-time provocative effect of diethylcarbamazine (DEC) on the circulation of microfilariae (mf) of nocturnally periodic Wuchereria bancrofti compared with the night mf count was evaluated in terms of sensitivity and specificity. In one test, the mean mf count for finger-blood samples from the same person on 3 successive nights was compared with the day-time post-DEC count; another test compared the mf count in one night-time finger-blood sample with the following day-time post-DEC count. Both tests were efficient (delta = 0.74 and 0.77, S.E. = 0.07 and 0.07, respectively). Night and day-time post-DEC mf counts were significantly correlated (r = 0.81, P less than 0.001, and r = 0.79, P less than 0.001 for the 2 tests respectively), but mf densities tended to be relatively lower after DEC provocation than in the corresponding night-blood samples. In both tests the regression coefficients differed significantly from zero (P less than 0.001). The simple 24h test was almost as efficient as the 3 nights test and based on its regression relationship, a table is presented predicting the expected night mf counts from the observed day-time post-DEC counts. The provocative dose of DEC, 2 mg/kg body weight, evoked no side-reaction in either microfilaraemic or uninfected persons. PMID- 3051543 TI - Factors affecting transmission of Wuchereria bancrofti by anopheline mosquitoes. 2. Damage to ingested microfilariae by mosquito foregut armatures and development of filarial larvae in mosquitoes. AB - Microfilariae (mf) of Wuchereria bancrofti from the midgut of 639 Anopheles gambiae, 557 An. arabiensis, 117 An. melas and 9 An. funestus were examined immediately after the mosquitoes had fed on carriers with different densities of mf. The percentages of mf damaged during ingestion were 57.1-60.0 in An. gambiae, 33.3-50.6 in An. arabiensis and 38.7-55.7 in An. melas. In each species the percentage of mf damaged was independent of mf density in the human host. A further 3657 An. gambiae, 2875 An. arabiensis, 347 An. melas and 32 An. funestus were examined 7 d or more after feeding on mf carriers. In An. gambiae and An. arabiensis, mean numbers of larvae per mosquito were strongly correlated to mf blood density, with similar regression slopes to those obtained from the regression of mf blood density on mean uptake of mf/mosquito. The ratio of mean numbers of larvae per mosquito to mean numbers of intact mf ingested per mosquito increased as the density of mf in the human host increased in An. gambiae and An. arabiensis, but decreased in An. melas as host mf density increased. PMID- 3051542 TI - Factors affecting transmission of Wuchereria bancrofti by anopheline mosquitoes. 1. Uptake of microfilariae. AB - Ingestion of Wuchereria bancrofti microfilariae (mf) from humans by 639 Anopheles gambiae, 557 An. arabiensis, 117 An. melas and 9 An. funestus was investigated. The mf densities in blood fell into 3 groups; 0-32mf/ml, 107-122 mf/ml and 421 1140 mf/ml. In An. gambiae and An. arabiensis percentage of mosquitoes ingesting mf was strongly associated with mf density in host blood; in An. melas the association was much weaker. Mean number of mf ingested per mosquito was also strongly correlated to mf density in An. gambiae and An. arabiensis but not in An. melas. At low mf densities both An. gambiae and An. arabiensis concentrated mf, with concentration decreasing as density increased. From regression analysis, observed and expected uptake of mf would be equal at 622.9 mf/ml in An. gambiae and 391.6 mf/ml in An. arabiensis. PMID- 3051545 TI - The changing response of Plasmodium falciparum to antimalarial drugs in east Africa. AB - For the past 20 years, chloroquine chemotherapy has been the single most effective malaria control measure in East Africa. The advent of chloroquine resistant Plasmodium falciparum has reduced the clinical effectiveness of chloroquine and this trend is likely to continue. Combinations of antifol drugs are at present effective inhibitors of most P. falciparum infections in the region, in spite of widespread resistance to pyrimethamine. The development of (i) sensitive methods for monitoring changes in sensitivity to antifol combinations, (ii) more effective and less costly alternatives to commercially available combinations, and (iii) investigation of host and parasite factors leading to drug treatment failure in P. falciparum infections has been the primary goal of the Wellcome Trust Research Laboratories in Kenya (directed by Dr W.M. Watkins) within the malaria programme of the Kenya Medical Research Institute, and collaborating laboratories at the School of Tropical Medicine and the University of Liverpool. PMID- 3051544 TI - Clinical and laboratory studies of malaria and melioidosis. PMID- 3051546 TI - Variability in drug susceptibility amongst clones and isolates of Plasmodium falciparum. AB - Heterogeneity within isolates of Plasmodium falciparum in regard to drug susceptibility is described from studies with three Thai isolates and some clones derived from them. One isolate (T9), which before cloning was resistant in vitro to chloroquine and pyrimethamine, contained a diverse assortment of clones, some of which were sensitive to one or other, or both, of these drugs. Another isolate (CH150) was initially sensitive to mefloquine in vitro, but, on recrudescence of infection in the patient, developed a number of clones all of which had diminished susceptibility to mefloquine. Drug pressure in a laboratory culture of CH150 resulted in a similar change in susceptibility. Hence resistant clones are thought to have been present as a minority component of the original isolate CH150. On testing uncloned isolates at different times after growth in culture, drug susceptibility showed considerable variability, but clones remained stable. Reaction in vitro of these isolates to some other drugs (amodiaquine, Fansidar, quinine) is also described, and the results are discussed in relation to changes in drug resistance of malaria parasites which may occur in laboratory cultures and under field conditions. PMID- 3051547 TI - Evidence of increased chloroquine sensitivity in Thai isolates of Plasmodium falciparum. AB - Sensitivity of Thai isolates of Plasmodium falciparum to chloroquine collected over the years 1978-1986 was measured by two methods: (i) by growth of previously cultured isolates for 72 h in presence of drug, and (ii) by the WHO standard in vitro microtest. During this period there were signs of a gradual increase in drug sensitivity, coinciding with the withdrawal of chloroquine for treatment of falciparum malaria in Thailand. PMID- 3051548 TI - The Plasmodium falciparum chloroquine in vivo test: extended follow-up is more important than parasite counting. AB - 349 in vivo tests of the susceptibility of Plasmodium falciparum to chloroquine, 25 mg/kg, were analysed. In some surveys, standard in vitro tests were also carried out. The proportions of sensitive and resistant infections in different areas found by the 2 methods were similar, but, within a given area, correlation between the two methods was often poor. Two RI cases and one RII/RIII case were sensitive in vitro, and it is suggested that the extended in vivo test may sometimes be more sensitive than the in vitro test, and that even in endemic areas, where reinfection is possible, patency on day 14 will nearly always be due to resistance. Parasite density data were analysed by calculating the geometric mean of each day's parasite density as a percentage of the day 0 parasite density + 0.1. Most resistant and sensitive infections attained minimal values on day 4, and it is proposed that assessment of sensitivity based on parasite densities should use day 4 values. Contrasts between materials were more clearly defined statistically when comparisons were based on ranking in vivo test classifications, than when based on day 4 parasitaemia. It is therefore suggested that, for epidemiological purposes, extension of tests to at least 14 d is more important than parasite counting. Parasitaemia above 20-25% of the day 0 value on day 2 in a severely ill patient, or persistent patency on day 4 in a symptomatic patient, are both indications for a change of treatment. PMID- 3051549 TI - Plasmodium falciparum incidence and patency in a high seasonal transmission area of Burkina Faso. AB - Using a mathematical model P. falciparum transition rates were calculated from the parasitological data collected from children of 2 villages of a high and seasonal transmission area in Burkina Faso (West Africa). During the rainy season, patent parasitaemia appeared every 40 to 60 d and was present for 60d approximately. The significance of these values is discussed. PMID- 3051551 TI - Studies in behaviour and malnutrition in Jamaica. PMID- 3051550 TI - Immunity to malaria in young Gambian children after a two-year period of chemoprophylaxis. AB - A cohort of 48 Gambian children was protected against malaria by fortnightly administration of Maloprim (pyrimethamine and dapsone) for 2 years between their 3 and 5 birthdays. A matched cohort of 47 children received placebo. During the year following the termination of prophylaxis there was no increase in the frequency of clinical attacks of malaria in the protected children compared with the control children. Antibody levels to circumsporozoite protein were measured by a radioimmunoassay and that to blood-stage antigens by a variety of techniques including an ELISA to whole blood-stage Plasmodium falciparum antigen, immunofluorescent assays (IFAT) to acetone fixed, glutaraldehyde fixed and unfixed parasites, a merozoite inhibition test and an opsonizing assay. Antibody levels were, in general, lower in protected than in control children and several differences between the two groups were statistically significant. When antibody levels were measured by ELISA and IFAT at the end of the following rainy season, when malaria transmission was intense, those in protected children had increased to comparable levels to those found in control children. Our findings suggest that chemoprophylaxis given for 2 years lowers malaria antibody levels but that it does not interfere with the development of protective immunity. PMID- 3051552 TI - Developmental time and mortality of larvae of Triatoma infestans infected with Trypanosoma cruzi. AB - Triatoma infestans were infected with Trypanosoma cruzi (zymodeme 1) at the first feed after eclosion from the egg; interstadial development times and mortality rates were then recorded until the imaginal moult and compared with those of uninfected controls. No retardation of development occurred in infected bugs and their mean total mortality rate (9%) was only slightly higher than that (6%) of uninfected controls (due solely to 4 additional deaths). This is the first demonstration, under optimal and standardized rearing conditions, that infection of T. infestans with T. cruzi has little or no effect on development times or mortality rates. PMID- 3051553 TI - The synergistic effect of low-dose cyclosporine and fluocinolone acetonide on the survival of rat allogeneic skin grafts. AB - Both CsA and topical FA can prolong the survival of skin allografts under the proper conditions. This study was performed to determine if there is a synergistic effect between these two compounds. Buf (RT1b) rat split-thickness skin grafts were transplanted onto the backs of Lew (RT1l) rats. The MST for the control group was 9.89 +/- 0.35 days. In rats given oral CsA, 2.5 or 5 mg/kg, daily from the second day of grafting, the MST was 16.0 +/- 1.9 and 13.6 +/- 0.4 days (blood CsA levels were 166 +/- 20 and 640 +/- 32 ng/ml at the time of rejection, respectively. Topical FA applied daily beginning 72 hr after grafting resulted in a MST of 24.1 +/- 3.6 days. When both topical FA and 5 mg/kg oral CsA were used, the allograft survival time was more than 100 days in 4 of 7 animals. When oral CsA 2.5 mg/kg was combined with topical CsA and FA, the allograft rejection was delayed until 50 days postgrafting in four of six animals. The synergistic effects of oral CsA and topical FA is significant, and thus allows for the use of a subtherapeutic dosage of each compound and provides a potentially safe means for prolonging skin allograft survival. PMID- 3051554 TI - Pancreatic secretory trypsin inhibitor as a marker for early detection of rejection in canine pancreas allotransplantation. AB - Serum pancreatic secretory trypsin inhibitor (PSTI)* was measured in the course of canine segmental pancreas allotransplantation without immunosuppression. Serum PSTI concentrations showed two distinct elevations: the first elevation was on the first day and the onset of the second elevation was the sixth day after operation. The first postoperative elevation of the serum PSTI level is thought to be related to the operative procedures, because the first elevation was observed after both autotransplantation and allotransplantation, and biopsies of the autograft and allograft at the first day after the operations showed nonspecific neutrophilic infiltration and no perivascular lymphoid infiltrates. The second postoperative elevation of the serum PSTI level is thought to reflect a rejection process because this elevation was not seen after autotransplantation, and biopsies of allografts at the sixth day after the operation showed typical perivascular lymphoid infiltrates and cellular rejection of the exocrine tissue. In addition, the onset of the second elevation of serum PSTI level preceded by about three days the onset of the elevation of blood sugar at the ninth day after the operation. The results suggest that serum PSTI can be used as a marker for diagnosis of early pancreatic allograft rejection. PMID- 3051555 TI - Pulmonary function of haplotype-matched and mismatched allografts in dogs treated with total-body irradiation, autologous marrow transplantation, methotrexate, and donor blood. AB - Seven beagle recipients surviving 2-11 years after allotransplantation of a left lung were available for study of pulmonary function. Significant reductions of ventilation and perfusion to the transplanted lung were documented by radionuclide scanning. These reductions in function were well-matched, however, and allowed relatively normal gas exchange, as measured by VD/VT, arterial PO2 and shunt fraction. The vasoconstrictor response of the transplanted lung to both hypoxia and stellate ganglion stimulation was comparable to that of the native lung. An abnormal rise in graft pulmonary vascular resistance and fall in PaO2 when the normal lung was made hypoxic suggest an inability of the transplanted lung to vasodilate and recruit blood vessels normally in response to increased blood flow. The animals were sacrificed at the conclusion of the pulmonary function testing. Pathologic study of the transplanted lungs showed minimal changes of rejection in spite of the fact that these recipient animals received no immunotherapy after the second posttransplant week. PMID- 3051556 TI - Hepatic reperfusion injury following orthotopic liver transplantation in the rat. AB - At 24 hr following orthotopic transplantation, rat liver grafts were perfused in situ for 7 min with trypan blue, a vital dye that provides information on hepatic microcirculation and stains nuclei of nonviable cells. Spotty and uneven dye distribution was observed indicating that hepatic microcirculation was disturbed 24 hr following transplantation surgery. Under these conditions, 15-20% of the hepatocytes were nonviable as assessed from trypan blue staining and frank necrosis. In contrast, perfusion of livers from untransplanted rats or liver explants exposed to cold ischemia for 60 min were judged normal by the criteria of uniform distribution of dye in the organ and absence of necrosis and nuclear dye uptake. Thus the observed damage was associated with reintroduction of blood and can therefore be classified as a reperfusion injury. The altered microcirculation and cell death following the operation was reduced markedly by perfusion of the cold, ischemic explant with nitrogen-saturated but not with oxygen-saturated buffer for 5 min prior to the implantation operation. Protection was even greater if the perfusion medium contained verapamil (20 micrograms/ml), a Ca++ channel blocker. We conclude that reperfusion of the stored liver causes an oxygen-dependent alteration in hepatic microcirculation that leads to hypoxia and scattered hepatocellular necrosis in the implanted graft. Brief perfusion of the hypoxic implant under anaerobic conditions may remove substrates involved in oxygen radical generation and prevent reperfusion injury upon introduction of oxygen into the graft via the blood. Taken together, these results suggest that removal of Euro-Collins' solution under anaerobic conditions may be beneficial clinically in preventing injury of surgical explants. PMID- 3051557 TI - Reliable indices for the determination of viability of grafted liver immediately after orthotopic transplantation. Bile flow rate and cellular adenosine triphosphate level. AB - One of the major problems accompanying liver transplantation is how to evaluate the viability of the grafted tissue at an early stage. The ability to assess immediate graft function would provide results useful in the determination of prognosis. The present study was undertaken to determine whether bile flow rates after liver transplantation were correlated with adenosine triphosphate levels and the survival of rats given transplants. In fresh-liver-transplanted rats, the one-week survival rate was 87%. The cellular ATP levels in the grafts decreased sharply prior to portal-venous declamping, but returned to nearly 80% of the normal level 4 hr after grafting, as did the total adenine nucleotide level and energy charge. When the grafts were subjected to warm ischemia for 15-min or 30 min periods prior to harvesting of the donor liver, the one-week survival rates decreased to 50% and 0%, respectively. In these cases, the levels of cellular ATP and bile secretion remained low and were proportional to the survival of the transplanted animals even 4 hr after transplantation. The relationship between the bile flow rates and the cellular ATP levels under various conditions revealed a good correlation, showing a saturation curve. The bile flow rates as well as the cellular ATP levels were therefore related to the survival rates of the transplanted animals. Thus it was shown in this experimental transplantation model that the monitoring of bile production after liver grafting is a useful indicator for assessing the extent of ischemic damage to the liver and for prognosis of the animal. PMID- 3051558 TI - Preservation of the canine liver for 24-48 hours using simple cold storage with UW solution. AB - The results of a series of 29 orthotopic liver transplants in the dog are described. The livers were preserved in a new cold storage fluid, UW solution, and were successfully transplanted after periods of storage of 24, 30, 36, and 48 hr. All six animals transplanted after 24 hr survived beyond 5 days after transplantation and had excellent graft function. Four of six survived for at least 5 days after 30 hr of cold storage, and five of five after 36 hr. Five of six consecutive dogs that received transplants that had been cold-stored for 48 hr survived for 5 or more days. This solution represents a substantial advance over all existing cold storage solutions for liver preservation. PMID- 3051560 TI - Plasma exchange for recurrent nephrotic syndrome following renal transplantation. AB - Patients with steroid-resistant nephrotic syndrome and focal segmental glomerulosclerosis (FGS) who develop end-stage renal disease are at risk for recurrence of the disease following renal transplantation. Recurrence of the nephrotic syndrome in renal allografts of two children with primary FGS was successfully controlled by plasma exchange. This report suggests that plasma exchange instituted early in the course of recurrent nephrotic syndrome may be beneficial in some patients with steroid-resistant nephrotic syndrome and FGS. PMID- 3051559 TI - Dermatologic lesions in a transplant population. AB - Many authors have reported an increase in the incidence of skin neoplasia in renal transplant patients. Two hundred and twenty-three patients, who received a renal transplants between 1965 and 1984, were examined for the presence of skin lesions. There was a high incidence of simple warts (24%) and hyperkeratoses (21%). Frank malignancy had developed in 9 patients; this is more than 4 times the expected incidence for the population of Northern Ireland. PMID- 3051561 TI - Megakaryocytic colony-stimulating activity in patients receiving a marrow transplant during hematopoietic reconstitution. AB - Megakaryocytic colony formation is dependent upon growth-stimulating activities present in human serum or plasma. Factors with diverse biological activities including megakaryocytic colony-stimulating activity (Mk-CSA) are provided by the medium of mitogen stimulated peripheral mononuclear cells or subsets of peripheral T cells. In this communication we describe the stimulatory activity of plasma collected from allogeneic and autologous bone marrow transplant recipients on the growth of megakaryocytic colonies. Mk-CSA was found to increase after transplantation as bone marrow regenerated. The stimulatory activities for CFU-M were greater in plasma from allogeneic bone marrow transplant patients receiving T cell-depleted donor marrow than in patients receiving unmodified donor marrow. Growth-promoting activities derived from mitogen-stimulated lymphocytes of T4 phenotype, a potent source of Mk-CSA, did not increase the frequency of CFU-M when cultured in plasma collected from transplant patients receiving a T cell depleted donor marrow. However, a further increase in the number of CFU-M was observed when exogenous Mk-CSA was added to the cultures supplemented with plasma from patients receiving unmodified donor marrow. Plasma of patients who received autologous marrow displayed similar Mk-CSA activity when compared with the activity of plasma obtained from transplant recipients receiving an unmodified allogeneic donor marrow. Stimulatory activities supporting multilineage colonies (CFU-GEMM), erythroid bursts (BFU-E), and granulocytic colonies (CFU-C) derived from plasma of the three different transplant groups revealed no statistical difference with respect to the frequency of CFU-GEMM, BFU-E, or CFU-C colonies when compared with pretransplant plasma. The results suggest that MK-CSA may play an important role in the regulation of megakaryopoiesis in vivo. Moreover the data suggest the presence of humoral regulators that appear to be different with respect to the processing of the donor marrow. PMID- 3051562 TI - Organ donation in three major American cities with large Latino and black populations. AB - It has been suggested that areas with large inner-city Black and Latino populations have worse organ donation rates than those with large suburban and rural White populations. Yet data are sparse. We studied family refusal rates (FRRs) to cadaver organ donation between 1/84 and 5/87 in three United States city-areas (New York, Los Angeles, and Miami) with large Black and distinct Latino populations. Blacks are at least 18% and Latinos at least 25% of the combined general population of the three cities, totaling over three and four million people, respectively. In addition, Blacks and Latinos represent 42% of cadaver transplant recipients, 49% of patients on waiting lists, and 57% of the patients on dialysis in the three cities. Combining the data from the three cities, Black (45%) and Latino (43%) FRRs were similar (P = .78), and each was significantly higher than that in the White population (17%) (P less than 0.0001). The overall refusal rate in NYC (42%) was significantly higher (P less than .0001) than in LA (26%) or Miami (21%), and LA's refusal rate was significantly higher than Miami (P = .03). The refusal rates for the White (31%) and Black (55%) populations in NYC were each significantly higher than their respective populations in LA (14% and 33%) or Miami (11% and 36%) (P less than .05). Although Miami Latinos had a lower FRR (35%) than Latinos in NYC (46%) or LA (45%), the difference was not statistically significant (P = .19 and P = .20, respectively). In the three cities combined, 515 of a possible 1772 medically and legally eligible organ donors were lost during the 40 months studied due to families' refusal of consent. This represents approximately 1000 transplantable kidneys and large numbers of extrarenal organs. Further studies are needed to elucidate the reasons for differences in donation rate among groups and regions in the United States. PMID- 3051563 TI - Elimination of leukemia cells from human bone marrow using floating beads. AB - A new in vitro immunophysical method of removing leukemia or lymphoma cells from autologous bone marrow is described. This new technique makes use of low-density polypropylene beads (density: 0.91) coated with a monoclonal antibody anti-CALLA (antibody ALB2). To ascertain its ability to selectively remove human B/pre-B hematopoietic cells, this technique was applied to normal human bone marrow cell suspensions contaminated with 1-5% of tumor cells. Samples were incubated with the floating beads at 4 degrees C on a rotating wheel for 60 min, followed by a 10-min decantation period, after which the beads bearing the tumor cells floated on the surface, whereas unbound normal marrow cells remained in suspension and were easily recovered free of beads. To demonstrate the feasibility of our method, 2 types of assays were carried out, one using target cell radiolabeled with 111indium, and the other a clonogenic assay. The first assays were to calibrate the different parameters (cellular density, quantity of beads, incubation time) with tumor cell lines: Namalwa (CALLA+) and Molt 4 (CALLA-). These 2 cells lines being able to clone, it is hard to envisage clonogenic assays. In this case, it is very hazardous to evaluate correctly the remaining clonogenic units of Namalwa cells. It is why radiolabelling assays were used for these first experiments. The second assays were to study a model close to the clinical setting and to control the safety of the beads on normal bone marrow cells. In this case, the mixture experiments in which only Namalwa cells were able to clone were evaluated with clonogenic assays, which are more sensitive than radiolabeling assays. A 3- to 4-log reduction of tumor load was achieved with 1-step treatment, and an average of 5-log depletion was obtained by repeating the process twice, as ascertained by the clonogenic assay. Viability, average recovery of nucleated cells, and stem cells potential following the purge were excellent. PMID- 3051564 TI - Cytokine-induced procoagulant activity in monocytes and endothelial cells. Further enhancement by cyclosporine. AB - IL-1, IL-2, and TNF alpha are important biological response modifiers of inflammatory and immunological reactions. Our experiments show that these cytokines are potent inducers of thromboplastin (TPL) activity but that their effects differ with regard to cell type and kinetics in human umbilical vein endothelial cells (HUVEC), monocytes (M), and mononuclear blood cells (MNC). Recombinant IL-1 alpha, rIL-1 beta and rTNF alpha all induced a dose-dependent increase in endothelial cell TPL activity, whereas rIL-2 had essentially no such effect. In the case of M and MNC cultures, IL-1 and IL-2 each induced TPL synthesis, IL-2 somewhat more slowly than IL-1. Special care was taken to exclude the effect of endotoxin present in the IL-1 preparations. Recombinant TNF alpha had a markedly smaller or no effect. When LPS was used to induce TPL synthesis, addition of rTNF alpha further enhanced HUVEC TPL, whereas no effect or a decrease in TPL was seen in MNC and M, especially in the presence of CsA and TNF alpha. Recombinant IL-1 beta also induced the synthesis of clotting factor VII in monocytes, thus allowing the formation of TPL-factor VII complexes, a most powerful trigger of blood clotting. IL-1 alpha, IL-2, and TNF alpha had no effect on the level of factor VII activity. Cyclosporine significantly augmented the level of TPL activity in HUVEC stimulated with rIL-1 alpha, rIL-1 beta, and rTNF alpha and in MNC and M stimulated with rIL-1 alpha, rIL-1 beta, and rIL-2. These actions of cytokines and cyclosporine may contribute significantly to the development of thrombotic reactions and fibrin deposits in transplanted organs, as well as to other pathophysiological pathways where activated clotting factors are involved. PMID- 3051565 TI - Effect of factors inhibiting HLA-DR antibodies before transplantation on kidney graft survival. AB - Blood transfusions administered before renal allografts are known to enhance graft survival. Among alternative hypotheses proposed to explain this effect, one of the most attractive is the possible induction of antiidiotypic antibodies directed against the specific antigen-binding site of donor-specific antibodies. In order to determine if such blocking antibodies are generated after blood transfusions, serial serum samples obtained before transplantation from 44 kidney recipients were analyzed for the development of HLA-DR alloantisera inhibitory activity by a microcytotoxicity inhibition assay. A significant correlation was found between the presence of inhibitory factors before transplantation and prolonged graft survival. However a clear relation between the development of inhibitory factors and the administration of transfusions could not be established. In addition the sera of 36 patients were studied for the presence of circulating immune complexes (CIC) before grafting. The presence of CIC was clearly associated with that of inhibitory factors, and with a prolonged graft survival. Thus these studies provide support for the development of blocking (possibly antiidiotypic) antibodies to anti-MHC in human renal graft recipients. PMID- 3051566 TI - Hyperacute allograft rejection mediated by anti-vascular endothelial cell antibodies with a negative monocyte crossmatch. PMID- 3051567 TI - Autoimmune neutropenia after renal transplantation. PMID- 3051568 TI - Effects of allostimulation and cyclosporine therapy on cytotoxic antibody production in highly sensitized prospective renal transplant recipients. PMID- 3051569 TI - Transmission and resolution of type I membranoproliferative glomerulonephritis in recipients of cadaveric renal allografts. PMID- 3051570 TI - Successful hepatic allograft procurement and transplantation from a donor after cholecystectomy. PMID- 3051571 TI - Neurotensin and vasoactive intestinal peptide levels during orthotopic liver transplantation in man. PMID- 3051572 TI - Acute humoral rejection after heart transplantation. PMID- 3051573 TI - Donor-specific transfusion and cardiac xenografts. PMID- 3051574 TI - Normalization of blood glucose levels in nondiabetic nude mice by human fetal pancreas after induction of diabetes. PMID- 3051575 TI - Hypothermic preservation of the rat liver assessed by orthotopic transplantation. III. Improved functional recovery with isotonic citrate solution and a stable prostacyclin analogue. PMID- 3051576 TI - International Symposium on Immunosuppression in Renal Transplantation. October 5 6, 1987, Barcelona, Spain. Proceedings. PMID- 3051577 TI - Renal transplants monitored with percutaneous biopsy and fine-needle aspiration cytology. PMID- 3051578 TI - Antirejection treatment with anti-thymocyte globulin in renal transplant recipients treated with cyclosporine as basic immunosuppression. PMID- 3051579 TI - Low-dose cyclosporine, anti-lymphocyte globulin, and steroids in first cadaveric renal transplantation. PMID- 3051580 TI - Three-year experience with cyclosporine and steroid double therapy in renal transplantation. PMID- 3051581 TI - Optimal results in cadaveric renal transplantation with low-dose cyclosporine and steroids combined with prophylactic anti-lymphocyte globulin. PMID- 3051582 TI - Low doses of cyclosporine, azathioprine, and prednisone in renal transplantation: immunosuppressive efficacy and reduced nephrotoxicity. PMID- 3051583 TI - The efficacy of low doses of cyclosporine A plus azathioprine. PMID- 3051584 TI - History of immunosuppression in kidney transplantation. PMID- 3051586 TI - Benefit of selective treatment with steroids in renal transplantation in various immunosuppressive regimens. PMID- 3051585 TI - Chronic therapy with cyclosporine A does not modify adrenal production of aldosterone in renal transplant patients. PMID- 3051587 TI - Acute renal failure after renal transplantation under various immunosuppressive regimens. PMID- 3051589 TI - Derivation of an algorithm for optimal initial cyclosporine immunotherapy in kidney transplantation. PMID- 3051588 TI - Collaborative transplant study data on efficacy of cyclosporine A in renal transplantation. PMID- 3051590 TI - Histologic findings in acute renal allograft rejection with triple drug immunosuppressive therapy: a retrospective analysis. PMID- 3051591 TI - T cell monitoring in kidney transplantation. PMID- 3051593 TI - Cyclosporine-drug interactions. PMID- 3051592 TI - Fractional excretion of sodium during acute rejection superimposed on acute tubular necrosis in the immediate post-transplant period. PMID- 3051594 TI - Chronic renal allograft rejection and cyclosporine. PMID- 3051595 TI - Pre-transplant transfusions in renal recipients treated with cyclosporine: a subject of controversy. PMID- 3051596 TI - Cadaver kidney transplantation in hyperimmunized patients. PMID- 3051597 TI - Sequential therapy with anti-thymocyte globulin and cyclosporine A in oligoanuric high-risk cadaveric kidney graft recipients. PMID- 3051598 TI - Use of a monoclonal antibody directed against interleukin 2 receptor in recipients of kidney allografts. PMID- 3051599 TI - Rejection treatment by polyclonal antibodies in kidney transplantation: reliable therapy without severe side effects. PMID- 3051600 TI - Cancers of unknown primary site: an enigmatic syndrome. PMID- 3051601 TI - Esophageal disease and angina pectoris. PMID- 3051602 TI - [Possible mechanisms regulating the entry of normal and neoplastic cells into the DNA synthesis phase]. AB - Literature data concerning some aspects in the regulation of the entry of normal and neoplastic cells into the phase of DNA synthesis are reviewed. Basing on the analysis of data reported by different authors, a hypothetical model is suggested for regulation of cell passing from G- to S-period. The main chain in the regulation is the constitutive synthesis of protein-repressor which blocks the expression of genes whose products are necessary for replication ("replication genes"). The repressor inactivation is achieved by growth factors through protein kinases. A comparison is made between the transcriptional mechanisms in eukaryotic and prokaryotic cells. Four hypothetic types of genes have been isolated responsible for regulation of progression of the cell to phase S. The role of quantitative and/or qualitative changes in the products of these genes in neoplastic transformation is discussed. PMID- 3051603 TI - [Instability of immunoglobulin expression during differentiation induction in the human lymphoblastoid cell line RPMI-6410t]. AB - The induction of immunoglobulin heavy chain classes switch from IgM to IgG was demonstrated in vitro in cells of RPMI-6410t line. The IgG+-sublines, formed as a result of the switch are characterized by instability of IgG synthesis. After removal of the inductor from the growth environment, IgG+ cells gradually reduce the level of secreted IgG. Such a transition to the functional rest state is likely to be connected with the convertible Ig-gene activity suppression in IgG+ cells, since after their activation by LPS IgG-secretion is partly or completely restored. The IgG+-sublines obtained may serve a convenient model for investigating the Ig-gene expression regulation in differentiated human B-cells. PMID- 3051604 TI - [In vitro movement of human embryonic fibroblasts with normal and anomalous sets of chromosomes]. AB - Migration of diploid postnatal and embryonic diploid and aneuploid cells was studied using a modified method of investigation of leukocyte migration under agarose. The method permits to study migration of fibroblasts and other proliferating cells. Embryonic fibroblasts were shown to move faster, than postnatal fibroblasts. Cells with trisomy 7, 9 and C, and triploid cells were found to move slower than diploid cells. Locomotor disturbances are supposed to be the basis of impairment of morphogenesis in chromosomal anomalies in man. PMID- 3051605 TI - [A method for obtaining histological sections from tissue previously studied by scanning electron microscopy]. AB - An accelerated method of paraffin embedding of tissue specimens previously examined with scanning electron microscopy is proposed aimed to obtain sections for routine histological examination. The tissue is passed through acetone, absolute alcohol, alcoholic-oil celloidin solution, chloroform to be eventually mounted into paraffin. The method allows obtaining good quality sections within 24 hours. PMID- 3051606 TI - Molecular biology: new hopes and challenges. PMID- 3051607 TI - Controlled clinical trial of a regimen of two durations for the treatment of isoniazid resistant pulmonary tuberculosis. AB - Patients with pulmonary tuberculosis who were failures of primary chemotherapy with strains resistant to isoniazid or to isoniazid and streptomycin were allocated at random to receive a regimen of rifampicin and ethambutol for 6 (4RE) or 9 months (7RE), supplemented in both treatment series by streptomycin plus pyrazinamide for the first 2 months. The patients were treated in hospital for the first 2 months and thereafter treatment was supervised on a daily basis in the nearest health institution by an appointed member of staff or at home by responsible members of the community. A total of 306 patients was admitted and 226 patients remained for analysis at the end of chemotherapy, 179 with a strain resistant to isoniazid alone and 47 with a strain resistant to isoniazid and streptomycin. There were only two failures at the end of chemotherapy, one in the 6-month series who had resistance to both isoniazid and streptomycin pretreatment, and one in the 9-month series who had resistance to isoniazid alone. For the 144 patients with initial resistance to isoniazid alone assessed up to 30 months, the relapse rates were low in both series: 4% for the 72 patients in the 6-month series and 3% for the 72 patients in the 9-month series. However, for the 34 patients with resistance to both drugs, three of the 14 in the 6-month but none of 20 in the 9-month series relapsed. PMID- 3051608 TI - Percutaneous drainage of tuberculous abscess of the psoas muscle. AB - Two cases of tuberculous abscess of the psoas muscle are reported which did not improve with antituberculosis chemotherapy. They were treated by sonographically guided percutaneous drainage through a pigtail catheter, with a successful result. PMID- 3051609 TI - Sternal abscess as a complication of BCG-revaccination. AB - A sternal abscess appeared 14 months after BCG revaccination of a 14-year-old girl. Culture taken from the abscess was positive for Mycobacterium bovis, which was identified as a BCG strain. PMID- 3051610 TI - Comparison of four digital maximum frequency estimators for Doppler ultrasound. AB - The performance of four methods for digitally estimating the maximum frequency waveform from the Doppler ultrasound spectrum, are described. The methods investigated are: a percentile method, D'Alessio's threshold crossing method [D'Alessio T. (1985) "Objective" algorithm for maximum frequency estimation in Doppler spectral analysers. Med. Biol. Engng and Comput. 23, 63-68.], a modified threshold crossing method, and a new hybrid algorithm. Evaluations of the variance and bias were performed using stationary simulated continuous wave (CW) Doppler signals of different bandwidths and signal/noise ratios (SNR) of 9 and 17 dB. Furthermore, a simulated nonstationary Doppler signal, similar to that from a normal internal carotid artery, was also used to compare the various methods. Overall, it was found that the modified threshold method and the new hybrid method have the best performance over a wide range of signal and noise hybrid method have the best performance over a wide range of signal and noise conditions; however, D'Alessio's method also performs well for low SNR's. PMID- 3051611 TI - A transmission line modelling approach to the interpretation of uterine Doppler waveforms. AB - An electrical analogue model of an artery that terminates into a vascular bed is presented. The model consists of an uniform transmission line that represents the artery and a load impedance that represents the vascular bed. The transmission line parameters are based on a well-established first-order approximation of the Navier-Stokes equations for fluid flow in distensible tubes. The model can be used to predict the incident and reflected components of both the arterial pressure and flow waveforms. In addition, it can predict the vessel diameter change and the mean blood velocity waveforms. In this study, the model was applied to the uterine artery so that the characteristics of the utero-placental circulation can be related to Doppler ultrasound recordings. It was found that the presence of the dicrotic notch in the uterine artery time-velocity waveform is the result of wave reflection and that a persistent notch past 20 weeks' gestation may be indicative of an abnormally high placental bed resistance. It is shown that the simulation results are consistent with the known physiological data and the clinically recorded uterine Doppler waveforms. PMID- 3051613 TI - An evaluation of the use of texture measurements for the tissue characterisation of ultrasonic images of in vivo human placentae. AB - A means of correcting for the depth dependence of co-occurrence features derived from ultrasonic images of the in vivo human placenta was derived. The corrected features were used to characterise the texture of the images. The features were found to correlate with a subjective assessment of the texture (p less than 0.001). Significant differences (p less than 0.05) between the placentae of smokers and nonsmokers at gestational ages 28 to 35 weeks were demonstrated. Also in this period, differences between patients who were normotensive and patients who became hypertensive were shown (p less than 0.05 for smokers, p less than 0.01 for nonsmokers). PMID- 3051612 TI - Comparison of theoretical scattering results and ultrasonic data from clinical liver examinations. AB - A theoretical analysis of soft-tissue ultrasonic scattering has been used to formulate specific results describing spectral parameters for tissue characterization. Results are applicable to clinical liver examinations. Three spectral parameters are mathematically expressed in terms of acoustic attenuation and the effective sizes, concentrations, and relative acoustic impedances of tissue scatters. Results from a clinical data base are shown to agree well with analytical results for each spectral parameter. Agreement is found for: spectral shapes; effects of attenuation; and correlations between parameters. Images of three spectral parameters are presented and their gray-scale features are evaluated with reference to analytical results. PMID- 3051614 TI - In vivo acoustic attenuation in liver: correlations with blood tests and histology. AB - Results of in vivo attenuation measurements in the liver have been obtained in 26 normal controls and in 51 patients with chronic diffuse liver disease. A modified real-time sector scanner was used for narrow-band amplitude attenuation examination. In the control group (people without apparent liver disease), a statistically significant correlation was found between acoustic attenuation in liver and two blood tests reflecting liver function: serum albumin (n = 24, r = 0.67, p = 0.002) and prothrombin time (n = 23, r = 0.63, p = 0.019). There was a statistically significant positive correlation between attenuation and fat for all biopsied patients (n = 51, r = 0.32, p = 0.023) and for patients with minimal fibrosis (n = 25, r = 0.45, p = 0.027). Although no correlation with fibrosis was found for all patients, in the group of patients with minimal fat there was a correlation with portal fibrosis (n = 33, r = 0.37, p = 0.035). This double blind prospective study shows that in the liver: (1) attenuation estimates appear correlated with clinical parameters (blood tests) in normal volunteers, and (2) large changes in fat affect narrow-band acoustic attenuation estimates to a greater degree than severe portal fibrosis in patients with chronic diffuse liver disease. Further research is needed before these estimates can become a clinical tool. PMID- 3051615 TI - In vivo and in vitro ultrasound beam distortion measurements of a large aperture and a conventional aperture focussed transducer. AB - Ultrasound focussing through human tissue of thicknesses varying from 10 mm to 35 mm has been measured for two transducers with diameters 50 mm and 19 mm both focussed at 50 mm (f/1 and f/2.6, respectively). Comparisons are made between the two-way focal depth beam patterns obtained in water and those obtained after passage through tissue to study the degrading effects of frequency-dependent attenuation and inhomogeneities, and their dependence on aperture size. The effects of frequency-dependent attenuation is to broaden the beam and shorten the focal distance. Inhomogeneities mainly increase the sidelobe levels and cause deviations from the central beam axis. A direct comparison of the beam patterns of the two transducers after passage through the same tissue samples shows that the resolution is improved by using the larger aperture. The use of the larger transducer in the in vitro measurements on three human liver specimens demonstrated an average improvement in the -6 dB beamwidth, over the smaller transducer, of 42% (standard deviation +/- 3%). The average improvement in the in vivo measurements on ten female breasts was 34% (standard deviation +/- 5%). The measured improvement in water was 52%. Therefore, the measured resolution improvement in tissue is approximately 2/3 of that obtained in water. The results indicate that for an f/1 transducer with a focal depth of 50 mm the upper limit of maximum useful aperture size has not been reached. PMID- 3051616 TI - Epidemiology of human exposure to ultrasound. PMID- 3051617 TI - Bibliography of biomedical ultrasound. No. 75. PMID- 3051618 TI - [Multiple sclerosis: an update]. PMID- 3051619 TI - Treatment of acute cellular rejection in renal transplantation patients on ciclosporin with antithymocyte globulin. AB - Our treatment of acute renal cellular rejection in patients on ciclosporin (cyclosporin A) involves the almost complete elimination of T cells (according to individual T cell monitoring in peripheral blood) through antithymocyte globulin (ATG) therapy. We present here the results of treatment of 44 histologically or cytologically proven acute rejection episodes in 33 of 62 consecutive renal transplantation patients. ATG therapy resulted in a 95.5% success rate when duration and dosage were individually adjusted according to T cell elimination in peripheral blood. Only two grafts were lost, both because of vascular occlusions. In neither case could it be clarified whether the loss was due to primary vascular lesions or the result of severe rejection or side effects of ATG. Our data show that the elimination of primed T cells with ATG permits the use of ciclosporin even in the presence of an established immune response. PMID- 3051620 TI - Ipsilaterality of motor innervation of canine urethral sphincter. AB - The functional activity of the sphincter muscle of the urethra is known to be controlled largely by the hypogastric and pudendal nerves. It remains unknown, however, whether innervation of the muscle by these peripheral nerves is ipsi- or bilateral. In an attempt to answer this question urethral closure pressure was determined simultaneously in the anterior, posterior, right and left portions of urethral wall in dogs. The pressure measurements were stereographed with the aid of a computer (stereo-UPP) and by this means the effect of unilateral section or electrical stimulation of hypogastric and pudendal nerves on the intraurethral pressure profile was analyzed. Unilateral section or electrical stimulation of the hypogastric nerve, distal to its division, produced a fall and a rise primarily in proximal intraurethral pressure, respectively, in all four directions. There was no significant difference in this response between the involved and uninvolved sides. Unilateral section of the pudendal nerve resulted in a fall primarily in distal intraurethral pressure in all four directions. No significant difference was present between the injured and noninjured sides. In contrast, electrical stimulation of the pudendal nerve distal to the point of its division caused a rise in intraurethral pressure in all four directions, with a significantly greater pressor response on the stimulated than on the nonstimulated side. These observations suggest decussating motor innervation of the urethra by the hypogastric nerves and also the possibility of the distal urethra being ipsilaterally innervated by the pudendal nerve. PMID- 3051621 TI - Role of Chlamydia trachomatis and mycoplasmas in chronic prostatitis. A review. AB - Acute bacterial prostatitis caused by common urinary tract pathogens is an infrequent disease, and diagnostic difficulties are rarely encountered. On the other hand, chronic prostatitis is a common disease requiring rather elaborate diagnostic procedures. We applied the localization protocol of the four-specimen technique and combined quantitative determinations of microorganisms and quantitative cytologic analysis plus, in chlamydial infections, serologic investigations. Our studies provide good evidence that Ureaplasma urealyticum and Chlamydia trachomatis must be considered etiologic agents in many cases of chronic bacterial prostatitis. These unconventional microorganisms are assumed to infect the prostate by way of intracanalicular ascension from the urethra. PMID- 3051622 TI - Microwave coagulation therapy for urinary bladder tumors. AB - A new device has been developed for microwave coagulation of urinary bladder tumors. Twenty-one patients with urinary bladder tumors were treated by irradiation with microwave energy of 2,450 MHz. Results were obtained as follows: (1) microwave coagulation was performed in 21 patients with transitional cell carcinoma of the urinary bladder. Excluding 4 patients who subsequently received radical cystectomy, 17 patients showed a complete response, although 2 patients subsequently developed recurrences in different parts of the bladder within the following several months. Histological examination of the excised specimen revealed complete eradication of the tumor in 2 patients. In the remaining 2 patients with high-stage tumor (T4), viable tumor cells were noted in the urethra or vaginal wall. (2) Although neither technical difficulties nor severe complications were encountered, transient urinary frequency and calcification of the bladder wall were noted. The results of this study indicate that microwave coagulation may be used in the treatment of both superficial and invasive tumors. PMID- 3051623 TI - [Continent reconstruction of an extreme urogenital sinus--case report of a so called high vesicovaginal confluence]. AB - The possibility of functional reconstruction as an alternative to urinary diversion is presented in a case of severe urogenital sinus with a high confluence between bladder and vagina and with urinary incontinence. The successive steps in the operation are described in detail, and the risks and complications are discussed. PMID- 3051624 TI - [Does sonographic kidney morphology and morphometry provide evidence of pediatric kidney function?]. AB - Compared with conventional X-ray investigation, ultrasound examination has a variety of advantages in the evaluation of urinary tract disorders in infancy and childhood. It does not involve exposure to contrast medium and radiation; low costs, minimal stress to the young patients, high diagnostic accuracy and easy reproducibility are further benefits. Postoperative follow-up of obstructive disorders can be easily performed and the results quantified by means of ultrasound. Morphometric data correlate well with renal function. As urologists become more familiar with ultrasonic morphologic and morphometric data, X-ray examinations in childhood will be confined to a few specific indications. PMID- 3051626 TI - [Results of single-stage repair of distal hypospadias using the King modification]. AB - From 1975 to 1986 34 patients undergoing repair of distal hypospadias between the age of 2 and 11 were explored. Standard method of the one-stage correction of distal hypospadias without or with moderate chordee was the operative technique according to the modification of King. After rolling the urethral groove into a tube the neourethra is covered with an asymmetrical part of dorsal hood. Thus the operative aims such as nonproblematic micturition, normal cohabitation and ejaculation and a good cosmetic result, avoiding psychological problems, can be obtained. The total complication rate was 23.4%; the correction of urethral fistulas in two layers as ambulatory operations was nonproblematic. By using Silastic foam the incidence of postoperative edema could be reduced significantly. PMID- 3051625 TI - [So-called nephrogenic adenoma (adenomatous metaplasia). A rare tumorous adenopapillary lesion of the urothelium in the urinary bladder in a 6-year-old boy following injury]. AB - A 6-year-old boy developed an extensive adenopapillary proliferation of the urothelium of the urinary bladder--a nephrogenic adenoma (adenomatous metaplasia) -2 years after he had been injured in an accident, which was followed by operation and medical treatment. Histologically this lesion is characteristic. Like other epithelial tumors of the urinary tract, it is very rarely found in children. A dysontogenetic or metaplastic pathogenesis has been suggested. Once more is known about them, adenomas may be diagnosed more often. The clinical picture and therapy are briefly discussed and a review of the literature is given. PMID- 3051627 TI - [Segmental unilateral priapism--a case report]. AB - A case of segmental priapism in a 24-year-old man with congenital spherocytosis is presented. Preoperative cavernosography and computer tomography clearly revealed thrombosis of the left proximal corpus cavernosum. Because the lesion had already been in existence for 5 weeks irrigation to flush out the thrombosis was no longer possible. PMID- 3051628 TI - [Combination hormone-chemotherapy versus hormone therapy alone in the initial treatment of advanced prostate carcinoma--a prospective cooperative study]. AB - The effectiveness of orchiectomy plus an antigonadotropic therapy alone (ethinylestradiol sulfonate), of orchiectomy plus a combined hormone and chemotherapy (ethinylestradiol sulfonate plus vincristine/vinblastine, cyclophosphamide, methotrexate) and of orchiectomy plus chemotherapy (vincristine/vinblastine, cyclophosphamide, methotrexate) alone for the initial treatment of advanced (stage IV) carcinoma of the prostate was compared. The cumulative 4-year survival rate was better after combined hormone and chemotherapy. Our results do not justify the administration of combined hormone and chemotherapy in previously untreated advanced carcinoma of the prostate. PMID- 3051629 TI - Renal cell carcinoma in cadaver donor kidney. AB - A cadaver kidney, reported to contain a "simple cyst," was received from another institution. Frozen section biopsy prior to transplant showed renal cell carcinoma. The contralateral kidney already had been transplanted at the other medical facility. PMID- 3051630 TI - Radioisotopic transit parameters in obstruction of pelviureteral junction. AB - Cortical and whole kidney radionuclide transit parameters were studied in 16 patients who underwent a pyeloplasty for unilateral pelviureteral junction (PUJ) obstruction. As expected, the whole kidney mean transit time was found to be very sensitive but, due to the low specificity, not very helpful in the distinction between obstructed and nonobstructed kidneys. The cortical transit reflected, in some cases, the favorable surgical outcome. In other cases, however, it could not be considered as a reliable parameter of the clinical status, since it was found to be normal preoperatively and prolonged postoperatively in patients whose parenchymal function improved significantly after surgery and who had their clinical symptoms relieved. PMID- 3051631 TI - Priapus and priapism. From mythology to medicine. PMID- 3051632 TI - [Epikeratophakia for the correction of aphakia]. PMID- 3051634 TI - [Effects of display-associated work on the visual organ]. PMID- 3051633 TI - [Surgical treatment of postburn symblepharon and pterygium]. PMID- 3051635 TI - William Hunting and The Veterinary Record. PMID- 3051636 TI - A century of letters to the editor. PMID- 3051637 TI - The pharmaceutical industry and The Veterinary Record. PMID- 3051638 TI - An editor remembers. PMID- 3051639 TI - The Royal College and The Veterinary Record. PMID- 3051640 TI - A matter of confidence? PMID- 3051641 TI - Use of a high frequency transducer with real time B-mode ultrasound scanning to identify early pregnancy in cows. AB - The reproductive tracts of 22 Friesian dairy cows were examined from seven to 35 days after insemination using a real time B-mode ultrasound scanner with a 7.5 MHz transducer. The earliest detection of pregnancy was at nine days when a vesicle was imaged within the lumen of the uterine horn. The early conceptus was seen at day 13 within the vesicle and these structures were followed ultrasonically until day 35. There was a sudden enlargement of the vesicle at day 19 and a heart beat was detected in the embryo at day 22. The allantois was imaged at day 23 and the amnion by day 29. The embryonic outline was clearly defined by day 33 when the body cavities could be discerned. This ability to determine pregnancy at an early stage should prove to be a useful technique in investigating the problems associated with early embryonic death in cattle. PMID- 3051643 TI - Prediction of calving date in beef cows by real-time ultrasonic scanning. AB - Three hundred pregnant beef cows between 35 and 125 days of gestation were scanned ultrasonically and their calving dates were predicted from measurements of fetal parts. The mean difference between the actual and predicted calving dates was 0.9 day with a standard deviation of 9.0 days. The accuracy and precision of the prediction of calving date were sufficient to be of benefit in the management of cows in late pregnancy and at calving. PMID- 3051642 TI - Effect of enrofloxacin therapy on shipping fever pneumonia in feedlot cattle. AB - The effect of enrofloxacin therapy was investigated in 110 male double-muscled cattle weighing 275 +/- 3 kg, during a spontaneous outbreak of shipping fever occurring 11 +/- 2 days after they arrived in the feedlot. Forty-six diseased animals were divided randomly into three groups A, B and C, containing 17, 19 and 10 animals, respectively; the animals in group A were injected intramuscularly once daily for three consecutive days with 2.5 mg/kg of enrofloxacin, those in group B with 5 mg/kg of enrofloxacin and those in group C with 10 mg/kg of oxytetracycline. Clinical, serological, production and respiratory functional observations were recorded. The animals were clinically cured after the three day treatment except for three in group A and two in group C. These five animals made a clinical recovery after a three day booster treatment with a dose of 5 mg/kg enrofloxacin. The changes in respiratory gas exchange values induced by shipping fever were completely reversed 15 days later, suggesting that there had been no irreversible lung damage. The daily weight gains and the arterial blood gas values of the three groups of treated cattle were not significantly different. The high efficacy of the low dosage of enrofloxacin in this clinical syndrome may be explained by its antibacterial activity against Pasteurella species and Mycoplasma species. This field trial supports the in vitro studies which suggested than enrofloxacin is an appropriate therapy in cases of shipping fever. PMID- 3051644 TI - Reaction to a hair shaft in the pulmonary artery of a dog. PMID- 3051645 TI - The role of endotoxin in the pathogenesis of coliform mastitis in sows. AB - Sows were made tolerant to Escherichia coli endotoxin by daily intravenous (IV) injection of the pyrogen. A refractory state was induced, characterised by a markedly decreased fever. In contrast, intramammary (IMM) infusion of only a quarter of the endotoxin dose to which the animals were made tolerant by IV injection produced a markedly increased fever. This finding suggests that inflammatory endogenous mediators were released in the mammary glands and that their subsequent absorption into the blood circulation, and not the absorption of endotoxin caused fever. PMID- 3051646 TI - The suitability of dogs as guide dogs for the blind: criteria and testing procedures. AB - Criteria and testing procedures with regard to the suitability of dogs as guide dogs for the blind were developed on the basis of a literature study and own observations. A profile of the guide dog comprising physical characteristics, skillfulness, behaviour, and obedience was drawn up. As a rule, the testing procedures concern health and skills of the dogs. In the skill test some elements of the behavioural and obedience test were included. The final evaluation is based on the results of physical examination and the skill test, unless testing of behaviour and/or obedience appears necessary as well. A method for evaluating the performance of the dogs as objectively as possible is described. Some implications of using and testing guide dogs are discussed. PMID- 3051647 TI - Spinal cord ependymoma in two young dogs. PMID- 3051648 TI - Rapid identification of Haemophilus somnus, Histophilus ovis and Actinobacillus seminis using the API ZYM system. AB - The API ZYM system, a commercially-available technique that measures bacterial enzyme activity was used to test 43 isolates identified as H. somnus, H. ovis or A. seminis and 19 from related genera. The enzyme patterns resulting from the API ZYM differentiated H. somnus and H. ovis from A. seminis and related genera but not from each other. An identification scheme based on 9 of the enzymes in the API ZYM and a few simple biochemical tests is proposed for the rapid and reliable identification of these bacteria in a diagnostic bacteriology laboratory. PMID- 3051649 TI - Double-sandwich enzyme-linked immunosorbent assay for determination of Escherichia coli heat-labile porcine enterotoxins. AB - A "double-sandwich" ELISA for the detection and measurement of a heat-labile enterotoxin produced by porcine enterotoxigenic strains of Escherichia coli (LTp) is described. In contrast with other heat-labile toxins, LTp did not bind to agarose gels and exhibited a very low affinity for GM1 in the classical GM1-ELISA technique. The similarity of LTp with cholera toxin was confirmed by immunoblotting. This property allowed the binding of LTp to rabbit IgG anti cholera toxin antibodies (covalently linked to polystyrene plates) and sheep anti cholera toxin serum. The immunocomplex was revealed by anti-sheep immunoglobulin antibodies conjugated with peroxidase. Application of the "double-sandwich" ELISA to the quantitation of toxin production by two strains, which differ only in the presence or the absence of the K88ab antigen, showed that the Ent+, K88+ strain produced significantly less toxin than the Ent+, K88- derivative. PMID- 3051650 TI - Effect of experimentally-induced villus atrophy on adhesion of K88ac-positive Escherichia coli in just-weaned piglets. AB - Three- to four-week-old, just-weaned piglets were infected with transmissible gastroenteritis (TGE) virus and the next day with K88ac+ enterotoxigenic Escherichia coli (ETEC). Histological examination of caudal jejunum and ileum of piglets killed 2-3 days after virus challenge (1-2 days after ETEC infection) revealed severe villus atrophy especially in the jejunum compared with controls (P less than 0.05). Four-5 days after TGE virus infection villus length increased and after 7 days it was near normal. Villi scraped from jejunal and ileal mucosa of the piglets were incubated in vitro with K88ac+ E. coli and the number of bacteria adhering to 250 micron villus brush border was counted. Attachment of bacteria to villi of piglets killed 2-3 days after TGE virus infection was significantly decreased in comparison with adhesion to villi of non-infected piglets or of piglets killed 7 days after the virus infection. Correlation between in vitro adhesion and villus height was 0.6649 (P less than 0.001). The results suggest that the experimentally-induced villus atrophy was attended with a temporarily diminished susceptibility of villus enterocytes to adhesion of K88ac+ E. coli. PMID- 3051651 TI - Mammalian renal modifications in dry environments. AB - The literature on the role of the kidney and renal morphological modifications in places of limited water supply is reviewed. The anatomical structures for urine concentration found in animals living in desert or arid environments, although not all occurring in one particular animal, are wide medullae, long loops of Henle, long proximal tubules, long collecting tubules, small renal corpuscles, extension of the renal pelvis, well developed elongated papillae, occurrence of giant vascular bundles, specialized ultrastructure of Henle's loops, epithelial changes in the collecting tubule, zonation of the vasa recta and peculiarity of the arterial supply to the kidney. The renal renin content is moderately high in these species. The renin-angiotensin-aldosterone system is very active, retaining Na+ with water. The urine is concentrated at the expense of other electrolytes. Both the renal blood and urinary flow rates are lower than in species with access to unlimited water supply. The juxtaglomerular apparatus components are topographically intimate for effective tubuloglomerular autoregulation of renal blood flow. PMID- 3051652 TI - Gastrointestinal food allergy and its role in large domestic animals. AB - The significance of food allergy as a primary cause for gastrointestinal disturbances in domestic animals, especially calves and piglets, is discussed. The immunological backgrounds and pathogenesis are described in some detail. The clinical and pathological manifestations in animals are related to those in man. Diagnostic possibilities, therapy and prevention, as far as known in animals, are mentioned and, based on human experiences, further extensions are proposed. PMID- 3051655 TI - [Possible use of positive pressure oxygen inhalation in chronic bronchopulmonary obstruction]. PMID- 3051654 TI - Herpes simplex virus glycoprotein D is sufficient to induce spontaneous pH independent fusion in a cell line that constitutively expresses the glycoprotein. AB - Spontaneous small polykaryocytes were detected in a cell line designated BJ-o that harbors the BamHI J fragment of herpes simplex virus 1 DNA and expresses constitutively glycoprotein D (gD). The fusion activity of BJ-o cells correlated with gD production and was drastically reduced following exposure of the cells to monoclonal antibody HD1 to gD. Studies on the characteristics and requirements of cell fusion dependent on gD led to the conclusion that the characteristics and requirements for gD-mediated fusion activity of BJ-o cells are similar to those previously reported for cell fusion induced by the virus in that (i) polykaryocytosis was not augmented by exposure to medium of low pH with or without prior exposure to trypsin, (ii) the number of polykaryocytes was reduced following removal of terminal sialic acid residues by neuraminidase, and (iii) the number of polykaryocytes was augmented by masking of high-mannose N-linked oligosaccharides with concanavalin A or with its reduced form, succinyl concanavalin A. This effect was reversed by competition with mannose. PMID- 3051653 TI - A survey of some drugs commonly used in the camel. AB - Specific recommendations for drug dosages for the camel are rare and doses for this species are usually extrapolated from those recommended for other species. The pharmacology and toxicity of drugs likely to be used in the camel needs to be further studied to ensure the efficacy and safety of these drugs in this species. Most of the reported work is on the chemotherapeutic efficacy of a few drugs long in use in other species against trypanosomiasis, mange and gastrointestinal nematodes. Areas of study most deficient are pharmacodynamics, pharmacokinetics and drug metabolism. The anatomical, physiological and biochemical peculiarities of the camel warrant more pharmacological and toxicological studies in this species. This article surveys the literature on the pharmacology, toxicity and therapeutic uses of some antiparasitic and antibacterial drugs and central nervous system depressants commonly used in the camel. It appears that camels are more susceptible to the toxic action of some trypanocidal drugs than other species. In certain cases they may metabolize some drugs differently. In general, the camel appears to be a good subject for analgesics and anaesthetics. PMID- 3051656 TI - [Unresolved problems of the medical support for the Armed Forces in World War II]. PMID- 3051657 TI - [Organizational and methodological problems of using computers in the dispensary care of the officer corps in a military polyclinic]. PMID- 3051658 TI - [Use of hyperosmolar blood substitutes for replacement in acute blood loss]. PMID- 3051659 TI - [Characteristics of the clinical picture and differential diagnosis of intestinal Proteus infection from Sonne dysentery and gastrointestinal salmonellosis]. PMID- 3051660 TI - [Reaction of the neuroendocrine system to surgical trauma to the ear]. PMID- 3051661 TI - [At the point in epidemic control protection for the troops]. PMID- 3051662 TI - [Epidemiology and prevention of malaria in a mountain desert locale with a hot climate]. PMID- 3051663 TI - [A method for the temporary arrest of external arterial hemorrhage from the vessels of the neck and head]. PMID- 3051664 TI - [Evacuation of the sick and wounded via inland water routes during World War II]. PMID- 3051665 TI - [Transplantation of bone marrow cells]. PMID- 3051666 TI - [Immunoenzyme method for demonstrating the antigen and antibodies to the human immunodeficiency virus and its use for the serological examination of different population groups]. AB - An enzyme immune diagnosticum for the detection of the antigen and antibody to human immunodeficiency virus (HIV) has been developed at D. I. Ivanovskii Institute of Virology of the USSR AMS. The method is based on the principle of competitive analysis using an antiviral conjugate which allows the sera to be tested without preliminary dilution and ensures a high sensitivity of the diagnosticum. The survey for HIV antibody covered 5743 subjects, among them 4898 Soviet citizens predominantly belonging to high risk groups of contracting AIDS infection, and 845 foreigners. No antibody to HIV has been detected in any of the Soviet citizens. Among the foreigners, antibody to HIV were detected in 6 subjects, all of them arriving from different African countries. PMID- 3051667 TI - [The low frequency of false positive results in the serological detection of infectivity with the human immunodeficiency virus in collectives of healthy young people]. AB - Among 1002 foreign students examined in Odessa 11 subjects with antibody to HIV, i.e. infected with HIV, were detected. A complete agreement of the results of the enzyme immunoassay and immune blotting test was observed. The reasons of this are discussed. A specific regional distribution of seropositive subjects by their permanent residence places was revealed. PMID- 3051668 TI - [Isolation and characteristics of monoclonal antibodies to the Pichinde arenavirus]. PMID- 3051670 TI - [Discussion of a new strategy for treating viral diseases]. PMID- 3051669 TI - [Adsorption study of the virions of the influenza virus and its individual proteins on polystyrene]. PMID- 3051671 TI - Virulent diverticulitis: a pitfall for the unwary. PMID- 3051672 TI - Uncle Sam needs nurses--one woman's response. PMID- 3051673 TI - Acetazolamide and dexamethasone in the prevention of acute mountain sickness. AB - We randomly assigned 32 healthy backpackers to receive placebo, acetazolamide (250 mg twice a day), dexamethasone acetate (4 mg four times a day), or both drugs in combination to determine the drug efficacy in preventing acute mountain sickness (AMS) at altitudes of 3,650 to 4,050m (12,000 to 13,300 ft). The incidence of AMS was high but symptoms were generally mild. Combined drug therapy was superior to both placebo and single drug therapy in risk reduction. Using acetazolamide alone was moderately beneficial in preventing the occurrence of AMS, although minor side effects were frequent. The use of dexamethasone alone did not significantly reduce the AMS incidence, and discontinuing its use resulted in symptoms suggestive of adrenal insufficiency. For recreational backpackers, routine drug prophylaxis is not recommended, in view of the mild nature of this illness and the adverse effects of medications. The efficacy of combined acetazolamide-dexamethasone therapy warrants further investigation at higher altitudes, where AMS is more severe, and the dexamethasone should be withdrawn gradually to avoid a possible adrenal crisis. PMID- 3051675 TI - Fasciitis and abscesses complicating liposuction. PMID- 3051674 TI - Therapeutic dilemmas in type II diabetes mellitus--improving and maintaining beta cell and insulin sensitivity. AB - These discussions are selected from the weekly staff conferences in the Department of Medicine, University of California, San Francisco. Taken from transcriptions, they are prepared by Drs Homer A. Boushey, Professor of Medicine, and David G. Warnock, Associate Professor of Medicine, under the direction of Dr Lloyd H. Smith, Jr, Professor of Medicine and Associate Dean in the School of Medicine. Requests for reprints should be sent to the Department of Medicine, University of California, San Francisco, School of Medicine, San Francisco, CA 94143. PMID- 3051677 TI - Arsenic poisoning. AB - This discussion was selected from the weekly Grand Rounds in the Department of Medicine, University of New Mexico School of Medicine, Albuquerque. Taken from a transcription, it has been edited by Ralph C. Williams, Jr, MD, Professor and Chair of the Department of Medicine. PMID- 3051676 TI - Superior sagittal sinus thrombosis. Still a killer. AB - Following treating a case of superior sagittal sinus thrombosis, we did an extensive search of the literature, eliciting 795 cases of the disorder. An analysis showed that even after the introduction of antibiotics, the preponderance of these cases have been diagnosed at autopsy. Our findings raise questions about the current methods of diagnosis and management of superior sagittal sinus thrombosis: Can the correct diagnosis be made earlier? Does a distinction between partial and complete thrombosis call for a different management? PMID- 3051679 TI - Medicare payment reform--a practitioner's perspective. PMID- 3051678 TI - Liver transplantation--the first 25 years. AB - These discussions are selected from the weekly staff conferences in the Department of Medicine, University of California, San Francisco. Taken from transcriptions, they are prepared by Drs Homer A. Boushey, Professor of Medicine, and David G. Warnock, Associate Professor of Medicine, under the direction of Dr Lloyd H. Smith, Jr, Professor of Medicine and Associate Dean in the School of Medicine. Requests for reprints should be sent to the Department of Medicine, University of California, San Francisco, School of Medicine, San Francisco, CA 94143. PMID- 3051680 TI - Building a consensus for physician payment reform in Medicare. The Physician Payment Review Commission. PMID- 3051681 TI - [Paul Wilhelm Heinrich Langerhans (1847-1888). In memory of the centenary of his death 20 July 1988]. AB - On occasion of the centenary of the death of Paul Langerhans on July 20th 1988, his life and career are reviewed and the importance of his discoveries is re emphasized. PMID- 3051682 TI - [Procorum]. PMID- 3051683 TI - [Colorectal cancer]. PMID- 3051685 TI - [Oncogene expression in colonic cancers]. AB - The expression of oncogenes plays a central role in the regulation of cell growth in normal and neoplastic tissue. Techniques developed recently (blot and in situ hybridization), enable the quantitative detection and topographical localization of oncogene expression at DNA and RNA level. In colonic adenomas and carcinomas and in non neoplastic mucosa of the large bowel, expression products of the oncogenes Ha-ras, Ki-ras, fos, and c-myc were detected. In most tumors a significant overexpression--as compared to the normal mucosa--of Ha-ras, Ki-ras, and fos mRNA was found. The overexpression of oncogenes, however, did not correlate with tumor progression, neither qualitatively nor quantitatively. According to recent studies (4, 7) it can be concluded, that the expression of the Ki-ras gene altered by a single point mutation, contributes significantly to the initiation and progression of epithelial neoplasias of the colon mucosa. It can be concluded, therefore, that the demonstration of oncogene expression products will play an important role in tumor diagnosis in the future. PMID- 3051684 TI - [Screening for cancer of the large intestine]. AB - A reduction of the high mortality figures from colorectal cancer is possible only when early diagnosis is achieved. This is the aim of annual screening of asymptomatic persons starting at age 45. The only practicable and effective screening method at present is searching for occult blood in feces with the Hemoccult-test. This test should therefore be part of the routine laboratory programme. PMID- 3051686 TI - [Immunologic processes in carcinogenesis in the intestine]. AB - It is hard to believe that the development of intestinal carcinomas is not linked -to some degree with a failure of the immunological defense mechanisms. For this several reasons may be evoked (for all exist examples) like a neutralization or blind-folding of the immune recognition, an inhibition of the effector cell activation or else genetic syndromes. New insights by a more pronounced research of the immune phenomena in the immediate tumor surroundings are needed. PMID- 3051688 TI - [Colorectal cancer--epidemiologic aspects]. AB - Since 1971 the incidence of large bowel cancer in Austria has been higher than that of any other neoplastic disease. In 1983 it affected 3778 men and women, representing 14.8% of all malignancies. Concrete suggestions for an improvement of data collecting for the National Cancer Registry and the installation of a National Polyp Study are made. In the eastern parts of Austria the risk for the development of colorectal carcinoma is twice as high as in the western parts of the country. Environmental risk factors are discussed. PMID- 3051687 TI - [Risk diseases for colorectal cancer]. AB - Among the risk conditions with an increased prevalence or incidence of colorectal carcinomas hereditary and acquired disorders have to be differentiated. While in hereditary forms new in vitro tests may play a role in the identification of patients at risk we have to rely on screening and follow-up programmes in the other group of patients. The intervals of these surveillance programmes should be acceptable to the patient since many risk groups seem to be questionable on a cost-benefit evaluation. PMID- 3051689 TI - [Colorectal cancer. Precancerous conditions]. AB - Colorectal precancerous stages are primarily benign lesions of the colon mucosa with a potential to malignant degeneration. They are macroscopically seen as polypoid lesions, but not only "adenomas" are to be seen as precancerous. Polypoid lesions which are not of neoplastic origin are not capable of malignant degeneration. Precancerous dysplasias are also found in the flat atrophic mucosa of chronically inflamed colon mucosa , which complicates the macroscopical judgement in the endoscopy. Problems also arise for the pathologist in the separation between inflammatory regenerative epithelatypia and high-grade dysplasia. PMID- 3051690 TI - [Colorectal cancer: classification and aspects of the proliferation kinetics]. AB - The exact tumor classification by the pathologist is the basis of adequate therapy of colorectal carcinomas. The classification includes the determination of the histological type of the carcinoma and the grading according to the criteria of the WHO and the UICC, as well as the staging according to the TNM system of the UICC and the Dukes classification. Most colorectal carcinomas are adenocarcinomas of tubular, tubulo-papillary and papillary subtypes. Mucinous adenocarcinomas are characterized by a pronounced extracellular mucus production. Signet ring cell carcinomas with intracellular mucus production are very rare and predominantly localized in the right-sided colon. Adeno-squamous carcinomas and squamous cell carcinomas are extremely rare in the large bowel. They are only mentioned for completeness. The histological grading proposed by the WHO distinguishes carcinomas of well (G1), moderately well (G2) and poor (G3) differentiation. Well and moderately well differentiated tumors can be regarded as carcinomas with low grade of malignancy, whereas poorly differentiated ones are carcinomas with high grade of malignancy. The new grading of the UICC distinguishes in addition to the well, moderately well and poorly differentiated carcinomas the undifferentiated tumors (G4). G1 and G2 correspond to low grade, G3 and G4 to high grade of malignancy. According to the 1987 nomenclature of the UICC-TNM system pT1 denotes tumor spread to the mucosa, or mucosa and submucosa, pT2 to the muscularis propria, pT3 into the subserosa or into nonperitonealized pericolic or perirectal tissue and pT4 a perforation of the visceral peritoneum or a spread into other organs.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3051691 TI - [The history of cardiology in Vienna]. AB - Established by Auenbrugger the physical diagnostic increased the special occupation with diseases of the heart. The "anatomical clinic" in Vienna developed it to a before anticipated exactitude. Later Nothnagel and Wenckebach payed special attention to the symptom of angina pectoris. Austrian pioneers of electrocardiography were Rothberger and Winterberg. PMID- 3051692 TI - [Angina pectoris]. AB - Coronary heart disease has many different clinical courses: it can cause rhythm disturbances, sudden death, pump-failure, no pain at all (silent ischemia) or typical angina. Heart-pain can occur "on demand" after physical or mental stress with a duration of 3 to 5 minutes with typical location and good response to nitrates. It also can cause atypical forms of angina such as angina on rest, mostly due to coronary spasms. Angina can stable over months and years but can suddenly increase in severity and duration. This form is called unstable angina, which has to be recognized as soon as possible since acute myocardial infarctions evolve rather frequently. Infarction is an irreversible myocardial damage but before it develops many measures can be taken to preserve the jeopardized myocardium. The recognition and differentiation of angina pectoris is therefore of utmost importance. PMID- 3051693 TI - [Thrombolytic therapy of acute myocardial infarct: which drugs? Which patients?]. AB - The efficiency of thrombolytic therapy in acute myocardial infarction has been documented in a large number of studies. The reduction of mortality has been shown for streptokinase (SK) and APSAC and various combinations (SK and aspirin). The recanalisation-rate is higher for rtPA as is the reocclusion-rate. Major bleeding complications are similar for all agents. However, the incidence of intracranial bleedings seems to be higher with rtPA; thus the cost-effectiveness for the tissue plasminogen activators is questionable. Attempts are made to focus on the implications of the major clinical trials for the therapeutic strategy in different patient-groups. PMID- 3051695 TI - Medicare: questions and answers. PMID- 3051694 TI - [Magnesium in cardiology. A challenge for new studies]. AB - Magnesium has been a controversial issue in cardiology for a long time. The following facts are now known: Magnesium plays a decisive role in the treatment of malign ventricular rhythm disturbances; this is especially true for Torsade pointes, which is connected with post-depolarisation and frequently causes sudden cardiac death. In the critical infarction phase intravenous magnesium therapy results in a reduction of malign rhythm disturbances and of the mortality rate. Magnesium plays a role in glycoside intolerance in patients with congestive heart failure. Magnesium deficiency is an important but rather neglected intermediary factor for the occurrence of (avoidable) side effects of renal, ototoxic and cardiac nature, emerge when using cytostatics, immunosuppressives and antibiotics. In several types of poisoning, but also in many other emergency situations with high levels of catecholamine (tetanus, phaeochromocytoma) magnesium can be used as an antidote. Clinically significant situations in which the role of magnesium has been either demonstrated or is still to be determined include diabetes (frequent deficiency demonstrated, significance for late complications under review), alcohol abuse (frequent deficiency of magnesium and its significance for rhythm disturbances demonstrated, correlations with other complications under review), tetanic syndrome, numerous psychiatric-neuromuscular disturbances, including generalized convulsive attacks, transitional syndromes, thromboembolic complications, lipometabolism disturbances, TIAS, PRINDS and loss of hearing. PMID- 3051697 TI - Medicare and nonparticipating physicians. PMID- 3051696 TI - State Medical Society of Wisconsin. 1988 membership directory. August 1, 1988. PMID- 3051698 TI - [Antimalarial activities: the last 40 years. Antimalarial action program]. AB - Around the turn of the century there were about 250 million cases of malaria each year and 2.5 million deaths. The introduction of DDT-type insecticides with residual effect around 1940 led to much more effective prevention than previously. WHO has been assisting Member States with malaria control ever since its foundation in 1948. In about 1950 it was proved that spraying DDT inside houses could interrupt transmission and lead to eradication within three years by exhausting the reservoir. Around the same time the discovery of the first cases of anopheline resistance to DDT introduced an element of urgency. The principle of eradication was adopted for the Americas by the Pan American Sanitary Conference in 1954, and for the world as a whole by the World Health Assembly in 1955, though it was acknowledged that the application of the principle to tropical Africa would be premature. With the aid of WHO, and under the leadership of its Expert Committee on Malaria, most control programmes outside tropical Africa were converted into eradication programmes. The results of the first 10 years of the global eradication programme (1957-1966) were spectacular but uneven; there were reverses due to financial, administrative or operational problems, or to the resistance or behaviour of the vectors, or to the inadequate development of basic health services. In Africa, the pilot projects generally failed to interrupt transmission. A new and more flexible strategy was adopted in 1969, whereby countries were invited to revise their programmes to take local circumstances into account. These revisions showed that in many countries eradication was not possible in the short term, but in the majority of cases they did not manage to put forward any genuine alternative strategy. Resources became increasingly difficult to obtain, whereas the cost of insecticides and transport went up with the price of oil. Research, which had been neglected for some time, once again became the order of the day and great importance was attached to malaria when the Special Programme for Research and Training in Tropical Diseases (TDR) was set up by WHO, UNDP and the World Bank. Since 1978 the emphasis has been on integrating malaria control with the primary health care system and on integrating health with development.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3051699 TI - Lung cancer risks of underground miners: cohort and case-control studies. AB - All underground mines have higher radon levels than are found in surface air. Ventilation is the primary method of controlling radon levels. Fourteen cohort and seven case-control studies done on underground miners are reviewed; they include many types of ore. Only five of the studies deal with more than 100 lung cancer deaths. Variations in the attributable risk are given. Some generalizations can be drawn from these studies: the longer the follow-up, the greater is the attributable risk, even though the relative risk is reasonably constant. The induction-latent period is quite variable but is shortened by high exposure rates, by cigarette smoking, and by increasing age at start of mining. The predominant histological type of lung cancer among miners changed from small cell undifferentiated for short follow-up time to epidermoid after long follow-up times. With short follow-up time, a multiplicative interaction between smoking and radiation was indicated, but, with long follow-up time, the two factors appear to be simply additive. This difference is probably due to the shortened latent period among cigarette smokers, not to synergism. PMID- 3051700 TI - Models for the analysis of radon-exposed populations. AB - Radon-222 is a radioactive decay product of radium-226 and uranium-238, which are found throughout the crust of the earth. Studies of underground miners clearly show that exposure to radon and its decay products increases the risk of developing lung cancer. Data on standardized mortality ratios from eight cohort studies indicate that the radon-lung cancer relationship is statistically homogeneous, even though cohorts are from different types of mines and from different countries. Regression methods for cohort data based on a Poisson probability model permit a thorough consideration of risk patterns. In this report, we review these methods, wherein the disease rate in each cell of a multi way table is modeled as a function of the cross-classifying variables. The National Academy of Sciences' Committee on the Biological Effects of Ionizing Radiation uses the Poisson regression approach to develop a model for age specific lung cancer risk which depends on cumulative exposure, age at risk, and time since exposure. This model is reviewed and its implications discussed. The most important determinant of lung cancer is cigarette smoking. This paper discusses relative risk models for analysis of joint exposure to radon and tobacco products. The review of available studies suggests that the joint relationship of radon and smoking with lung cancer is consistent with a multiplicative model, but a submultiplicative relationship is most likely. An additive model is rejected. PMID- 3051702 TI - Pathophysiological changes of the cochlea during ageing--a review. AB - Research of the pathophysiology of the age-related human presbycusis was largely initiated by those investigations in which the cochlea's damages were experimentally induced by various acoustic exposures. It became apparent that the outer hair cells are more vulnerable to different noise influences than the inner hair cells. On the other hand, the role of the altered ultrastructural elements of various types, cannot be ruled out. The evolving of the age-related presbycusis was to a great extent established using large-scale model experiments, including mainly rodents. It seems that the age-bound presbycusis (cf. intrinsic degeneration of the cochlea) begins at the apical regions which will be later more worsening by various extrinsic traumas. It is suggested that the damages gradually taking part in the inner ear structures are bound to the age-dependent processes in which the ever-increasing formation of cross-linkages plays an important role as ageing progresses. PMID- 3051701 TI - A policy to control the spread of HIV infection. AB - Prevention of transmission of HIV infection is the most important public health concern of the AIDS epidemic. To date, unfortunately, we have failed to contain the epidemic. The increasingly rapid spread of HIV into the IV drug-abusing population and subsequent heterosexual transmission represent a further failure of the public health system. Current organization of the public health programs, especially the lack of independence and adequate financial and personnel support, is an extremely serious problem. More funding may not be the answer, unless there is better organization. Identification of infected individuals and a vigorous education program must be implemented. HIV antibody-positive individuals should be followed carefully in order to evaluate the risk factors for AIDS and efficacy of specific interventions. PMID- 3051704 TI - [From a regional medical foundation to a district hospital--on the construction history of the Zwickau hospital]. PMID- 3051703 TI - [Significance of the "life satisfaction" concept for gerontology]. AB - Issuing from perceptions of the occupational psychology and -sociology about the ambiguity of the category "satisfaction" the author derives the demand for more subtle recording of "satisfactions". The ideological background of growth in importance of the category of "satisfaction" in the bourgeois social gerontology research is referred to the same way as the statement that "satisfaction" no is an absolute (primary) goal of gerontological efforts. PMID- 3051705 TI - [Pregnancy conflict counseling]. PMID- 3051707 TI - [Diagnostic viewpoints in meningitis and encephalitis in adulthood]. PMID- 3051706 TI - [Prevention of thromboembolism with heparin and dihydroergotamine (DHE) in internal medicine--comparison of various administration forms of dihydroergotamine]. PMID- 3051708 TI - [Therapeutic viewpoints in meningitis and encephalitis in adulthood]. PMID- 3051709 TI - [Human immunodeficiency virus (HIV). 1: Transmission and risk of infection]. PMID- 3051711 TI - [Clinical manifestations of HIV infection in the oromaxillofacial area]. PMID- 3051710 TI - [Human immunodeficiency virus (HIV). 2: HIV infections caused by accidental exposure in medical personnel]. PMID- 3051712 TI - [Staphylogenic Lyell syndrome]. PMID- 3051713 TI - [Viral infections in pregnancy and their effects on the newborn infant and the infant]. PMID- 3051714 TI - [Botulism--clinical aspects, diagnosis, therapy]. PMID- 3051715 TI - [Endemic and tropical helminthiases]. PMID- 3051716 TI - [Prevention, diagnosis and therapy of significant important tropical diseases]. PMID- 3051717 TI - [Tachycardic arrhythmias--references for the procedure in ambulatory care]. PMID- 3051718 TI - [Emergency diagnosis of acute cerebrovascular insufficiency]. PMID- 3051719 TI - [Emergency therapy of acute cerebrovascular insufficiency]. PMID- 3051720 TI - [Phlebothrombosis]. PMID- 3051721 TI - [Coma as the leading symptom]. PMID- 3051722 TI - [Fever--infection]. PMID- 3051723 TI - [The 125th birthday of the roentgen pioneer Max Levy-Dorn (1863-1929)]. PMID- 3051724 TI - [Principles of allergic reactions]. PMID- 3051725 TI - [Pathogenic mechanisms of asthma and bronchial hyperreactivity]. PMID- 3051726 TI - [The diagnosis of bronchial asthma]. PMID- 3051727 TI - [Drug therapy of bronchial asthma]. PMID- 3051729 TI - [Crisis asthma and anaphylactic shock]. PMID- 3051728 TI - [Allergy to insect venoms]. PMID- 3051730 TI - [Pseudoallergic reactions]. PMID- 3051731 TI - [Exogenous allergic alveolitis]. PMID- 3051732 TI - [Allergic diseases of the nose, ears and throat]. PMID- 3051734 TI - [Nonmedicamentous therapy of asthma]. PMID- 3051733 TI - [Atopic diseases of the skin]. PMID- 3051735 TI - [Organization and occupational health monitoring of army personnel]. PMID- 3051736 TI - [Some aspects of the complaint problem in occupational medicine]. PMID- 3051737 TI - [HIV (human immunodeficiency virus) and HIV-associated diseases]. AB - The discovery of the Human Immunodeficiency Virus (HIV), the characterization of its molecular biology and the development of serologic methods for detecting antibodies have led to a better understanding of HIV-associated clinical syndromes. Recently, the Centre of Disease Control has proposed a classification of HIV-related conditions. This classification forms the basis for this review. It is completed by remarks on antiviral and immunomodulating drugs and its effects on HIV. Difficulties in development of vaccines are discussed. PMID- 3051738 TI - [Mechanical emergency stimulation in asystole and extreme bradycardia]. AB - Repetitive precordial thumping is the simplest method of temporary cardiac pacing. In 90 patients out of 100 with witnessed cardiac arrest because of asystole or marked bradycardia the critical situation could be effectively bridged over by this until a sufficient spontaneous rhythm returned or electrostimulation was ready to function. 69 patients were conscious during the stimulation. During the mechanical pacing only in 2 patients there occurred ventricular flutter or fibrillation, which was stopped by electric defibrillation. The existence of the myocardial contractility is the presupposition for effective mechanical and electrical pacing. PMID- 3051739 TI - [Russia and Halle reciprocity in medicine of the 18th century. Correspondence, scientific communication and reception]. AB - In the history of the interrelations of Halle and Russia in medicine during the 18th century Francke's Foundations, the Faculty of Medicine and the Academia Naturae Curiosorum are important institutions. The correspondence of these three institutions with the partners in Russia became remarkable for the scientific communication and reception. The activities of the working teams which were embedded in these authoritative bodies were scarcely inferior in importance to the centres of enlightenment of that time working in other towns (for instance in Leipzig). In the field of medicine Halle occupied a leading position for the early knowledge about Russia. PMID- 3051740 TI - [Halle physicians in the pre- and early history of modern endocrinology]. AB - Textbooks of endocrinology separate the modern development (since 1920) from a pre- and an early history of this discipline. Researching facts of the universities of Wittenberg and Halle appear in the lists of the two periods of inauguration. Pioneers as Ernst Joseph lesser, Ernst laqueur and Frigyes Verzar place the university of Halle in the range of early descriptions of endocrine data. PMID- 3051741 TI - [The functional pathology of hypophyseal adenomas]. AB - This review summarizes some aspects of pituitary adenoma pathology. A new embracing pituitary adenoma classification has been developed which correlates morphologic findings with endocrine activity. It is based on hormone content, histologic, immunohistochemical and ultrastructural features, cellular composition and cytogenesis, and separates pituitary adenomas into 7 distinct entities. PMID- 3051742 TI - [What is appropriate in postmenopausal osteoporosis?]. AB - The term of postmenopausal osteoporosis created 1940/1941 by Albright and his coworkers corresponds on our days no more to the original conception, because it is used now in a broader form. The resulting practical consequences, e.g. the postulates of a general medical prophylaxis, induce a new discussion which is adapted to the scientific state of to-day and the real importance. PMID- 3051743 TI - [Ludwig Haberlandt (l885-l932) and the development of hormonal contraception]. AB - Ludwig Haberlandt, physiologist in Innsbruck, tried in the late twenties to develop hormonal contraceptives based on sexhygienic ideas of Sigmund Freud. Although the chemical-physiological knowledge of that time encouraged this plan, after Haberlandt's death in 1932 his tests were dropped and forgotten. It was after World War II, when research in this field was begun in the USA by Gregory Pincus, supported by "Planned Parenthood Federation" under leading of Margret Sanger. Although clinical tests proved to be successful, restrictive social political conditions in the fifties delayed this project considerably. But in 1958 the first Anti-Baby-Pill reached the US-market. PMID- 3051744 TI - [Recommendation for the classification of male hypogonadism]. AB - For the adequate therapy and prognosis the division of the forms of male hypogonadism is of practical significance. The clinical picture is determined by 4 factors (dimensions): 1. On the basis of the vertical dimension hypogonadism can be hypothalamic, hypophyseal, testicular or peripheral in its origin; 2. Depending on the impaired testicular functions (single or both), hypogonadism can be selective or total; 3. Depending on the severity of the lesion hypogonadism can be complete or incomplete; 4. the time-factor indicates the beginning of the lesion (pre- or postpuberal). The duration of the lesion is also of clinical significance. For the division of male hypogonadism, two etiopathogenetical groups can be suggested: Group I: "Classical" male hypogonadism. Hypothalamus hypophysis-testis system primarily damaged; Group II: "Consecutive" male hypogonadism. Hypogonadism induced by other diseases. PMID- 3051745 TI - [Endocrinology from the viewpoint of the zoologist]. AB - The argument is presented that the scientific discipline of endocrinology originated in experimental zoology. The identification of the first ductless glands and the characteristization of their products required the proof that their excision caused recognizable changes which could be reversed by replacement therapy. This could only be obtained from animal experimentation. Zoologists contributed, but an important factor was the tradition of zoology in the medical curriculum, especially in Middle Europe from where sprang the immortal names of Berthold, Langerhans, Knauer, von Halban, Fraenkel, Biedl, and many others. PMID- 3051746 TI - [Knowledge of endocrine disease pictures from the viewpoint of the pathologist]. AB - Endocrinology is an independent discipline based on biochemistry and pathochemistry, but we cannot know any endocrine disease entity without making pathomorphological research. In these days professional pathologists cannot define any new endocrine disease entity by the means of the macroscopical examination, but they may contribute to the micromorphological confirmation of many syndromes using the results and methods of histochemistry, topochemistry, immunofluorescence and isotopic diagnostics. In order to give an acceptable explanation they have to integrate the ultrastructural picture with the classic cytologic picture. This the unity of the functional and morphological aspects can prove to be especially lucrative in the research of endocrinology. PMID- 3051747 TI - [Light microscopic investigation of the tissue integration of Lyodura soft implants]. AB - Commercial preparations of human "Lyodura soft" were implanted into the subcutaneous connective tissue of 40 Wistar rats. After survival times from 7 days up to 1 year the samples were examined by light microscopy. An initial reaction of mononuclear cells, activated fibroblasts, and some giant cells of foreign body type in the surrounding connective tissue is followed by the development of a fibrous capsule rarefied by cells around the implant in the time up to 3 months. After 6 months and more clearly after one year the capsule is reduced or even completely vanished. The implant remains then integrated in the subcutaneous connective tissue almost without residual reactions or signs of resorption. PMID- 3051748 TI - [Investigations on the tissue compatibility of silicon rubber materials using experimental animals]. AB - The reaction of subcutaneous tissue in guinea-pigs at implants of six kinds of silicone rubber materials (VEB Chemiewerk Nunchritz) was investigated by semiquantitative and quantitative methods. These based on the "Programme to the Assessment of Subcutaneous Tissue Reactions at Implants" by Knofler, Keller etc. The histocompatibility of the tested materials can be compared with that of titanium. One of the silicone rubbers differs from both the reference material and the other silicone rubber materials significantly and is assessed to be more suitable than the alloy Gisadent KCM 83 (chromium cobalt molybdenum alloy). PMID- 3051749 TI - [Demonstration of dual liver systems after experimental auxiliary partial liver transplantation using the alternative of sequential hepatobiliary scintigraphy]. AB - Hepatobiliary sequential scintigraphies by 99m Tc-ROTOP-EHIDA were carried out for judging the dual liver system after experimental auxiliary transplantation of a part of the liver. A functional advantage of the host liver in contrast to the transplant was found in all cases investigated. Ischemic transplant damages could be detected in all experiments and were scintigraphically recognised at an early stage. In addition to transplants with a functioning entire parenchyma, the investigations showed transplants with functionally active parenchyma zones and such ones without any functional evidence. A quantifiable determination of the global functional capacity of the hepatobiliary system of both livers was possible and also the evidence on residual functions. The different behaviour of the kinetics of the radio-pharmacon with normal function and in case of hepatic ischemia is discussed. PMID- 3051750 TI - [Education for the elderly]. PMID- 3051751 TI - [Education and life long learning]. AB - In this article the term "lifelong learning" is discussed in detail from a pedagogic perspective. Lifelong learning is seen as an integral part of "competence" and of mastering different developmental tasks in old age. Moreover, it is seen as a component of "education". Education comprises not only the acquisition of knowledge, but is understood to develop strategies and competence for coping with tasks and crises. This is illustrated in discussing theoretical models of critical life events and developmental tasks. PMID- 3051752 TI - [Senior studies--a model of retirement scientific continuing education for elderly adults]. AB - Against the background of a changing situation of "older adults" within a changing society, and proceeding from the historical development of "Opening the Universities to Elderly People" in the Federal Republic of Germany, the university-specific objective of studies for elderly people at university was worked out. It is the universities' task to develop education models and prototypes for cooperation with elderly people within the field of applied education and socialization research for tertiary socialization (sociogeragogy). This kind of study, as further education for elderly persons, is closely connected with teaching and research. Aspects of education and socialization reflecting the progress of the elderly persons, as well as methods of such educational work, explain this connection, including the development of a transpersonal identity. The task of the European cooperation of those universities which are working on such programs in order to open the universities to elderly people, is also discussed. PMID- 3051753 TI - [The elderly and new media]. AB - This article discusses the importance of using new communication technics for the quality of life in old age. Different models of new technics and the consequences for daily life in old age are illustrated. Using new communication technics in old age creates new tasks for gerontology. These scientific and practical tasks are discussed, too. PMID- 3051754 TI - [The Tree and Figure Test as a diagnostic aid in geriatric patients]. AB - The importance of the "Baumtest" is discussed in analyzing the special way in which patients experience their illness and the loss of psychological and physiological functions. The investigation affirms the hypothesis that the pictures reflect patients' attitudes towards the illness and damage to functions. This test is recommended as an instrument for diagnosing the psychological state of the geriatric patient. It is necessary, however, to complete the diagnosis by means of other instruments and by exploration. PMID- 3051755 TI - [The thyroid gland and the breast]. AB - The results of experimental clinical and epidemiological studies published so far from 1896 until today on the connection between thyroid and breast diseases doubtless require critical examination, since these results are highly contradictory. Without being able to go into details of the sources of methodological errors and the highly heterogenous investigation material, two pathophysiological mechanisms can be discussed in the investigation of the interrelationship between the thyroid and the breast: 1. A low level of thyroid hormone might make the breast hypersensitive to prolactin, which might induce dysplasia or neoplasia of the mammary epithelium. In addition, there is the almost identical neurohumoral regulation of the hormones of the thyrotropic and lactotropic cells in the anterior pituitary. 2. Thyroid hormones have an effect on peripheral androgenic and estrogenic metabolism. The hyperthyroid state may cause an increase in the concentration of sex hormone-binding globulins, which might give rise to an alteration of the beneficial effect of sex hormones on the cellular levels. Compared to this, the iodine deficit might lead to neoplastic transformation of the mammary epithelium via the enhanced gonadotropin stimulation and the subsequent chronic hyperestrogenism. For better understanding of the always topical problem of the not yet completely elucidated correlation between thyroid function and breast diseases, a two-track procedure is appropriate: 1. Thyroid receptor assay on animal and human breast tumor tissue and 2. prospective clinical studies on a large scale.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3051756 TI - [Prolactin--significance in benign and malignant breast diseases]. AB - At present, the role of prolactin has not yet been precisely determined both in the development and in the progression of benign and malignant diseases of the breast. Experimental investigations and data of the human receptor assay have clearly indicated the initial and promoting activity of prolactin in the genesis of breast tumors. However, the data published so far, which is in some cases highly contradictory, does not allow a positive appraisal on the correlation of the prolactin level and breast diseases in clinical research and experimental studies. The studies still to be carried out in human medicine with precise definition of the TRH-prolactin-target cell axis compared with the proof of the receptor radioimmunoassay in healthy and malignant breast tissue which has still not yet been provided might give rise to rather more clarity in this rather manifold problem. PMID- 3051757 TI - [Clinical introduction to aneurysmatic bone cysts]. AB - The aneurysmal bone cyst is according to the description of Jaffe and Lichtenstein (1942) a histomorphological entity. It is a benign centric or eccentrically localized tumor like lesion with preference to the metaphysis. 75% of all aneurysmal bone cysts occur within the first two decades of life. On x-ray the diagnosis is not always clear. In differential diagnosis juvenile bone cyst, giant cell tumor, chondroblastoma, chondromyxoid fibroma, non ossifying fibroma, fibrous dysplasia and telangiectasic osteogenic sarcoma have to be considered. Final diagnosis requires histological evaluation. Curettage or resection with bone grafting is the predominant therapeutic procedure. Radiotherapy might cause malignant degradation and should only be used in elected cases. For propaedeutic evaluations of aneurysmal bone cysts 851 cases in world literature are reported. PMID- 3051759 TI - Early results of a new heart transplant program in Virginia. PMID- 3051758 TI - [Comment on the publication "Sonographic follow-up of hip dysplasia with expanding trouser therapy"]. AB - It should be pointed out that hipsonography by application of a flexible control procedure is not only suitable for introduction of therapy for CHD at an early stage, but also for avoiding superfluous treatment. Secondly, the authors are of the opinion that universal screening for CHD by means of hipsonography is for financial considerations not justifiable. PMID- 3051760 TI - Cardiac transplantation in 1988. PMID- 3051761 TI - [A comparative study of the single knot and continuous suture technic of the aorta of the growing rat]. PMID- 3051762 TI - [Effect of naftidrofuryl on the ischemia model in healthy probands]. PMID- 3051763 TI - [Dissection of the internal carotid artery--an oft-mistaken disease picture? Case report and review of the literature]. PMID- 3051764 TI - [Autotransplantation and extracorporeal repair of a renal vascular abnormality in a patient with a single kidney]. PMID- 3051765 TI - The hammer syndrome. Trauma-induced lesion of the ulnar artery. PMID- 3051766 TI - [Means for evaluating the contractile capacity of the heart and heart muscle]. PMID- 3051767 TI - [The role of the hepatic stroma in the pathogenesis of hepatitis]. PMID- 3051768 TI - [Characteristics of the action of synthetic polyampholytes on the capsules of pathogenic and cultured Treponema pallidum]. PMID- 3051769 TI - [AIDS: the dermatological aspects of the problem]. PMID- 3051770 TI - [Differential diagnosis and treatment of patients with bullous dermatoses]. PMID- 3051771 TI - [Hippocrates on skin and venereal diseases]. PMID- 3051772 TI - [The Rebuck skin window method in the differentiated assessment of the activity of phagocytic reactions in patients with foot mycoses and eczema]. PMID- 3051773 TI - [Morphology and function of the human spleen. Enzyme histochemical and electron microscopy studies of the splenic lymphatic vessels, nerves and connective tissue structures]. AB - Many questions concerning the morphology of the spleen have been cleared up in the last 20 years by the application of new methods of investigation, especially electron microscopy and enzyme histochemistry. With but a few exceptions, however, only the splenic parenchyma (the red and white pulp) were studied. Such special structures of the human spleen as nerves, lymphatic vessels and their supporting tissue, which may play an important role in the coordination and integration of the different functions of the white pulp (secondary lymphatic organ) and the red pulp (blood filter), were hardly ever studied with modern techniques. Investigating these structures light and electron microscopically and enzyme histochemically it was attempted to complete our present knowledge of the histology of the human spleen. In addition, by comparing the study of special altered spleens with experimental data it was attempted to clarify the importance of these structures for the physiology and pathology of the spleen. A total of 151 normal and pathologically altered spleens from the bioptic and autopsy material of the Pathological Institute of the University of Kiel were examined. In addition to conventional light microscopy the spleens were investigated enzyme histochemically and cytochemically, fluorescence microscopically and electron microscopically. The following enzyme reactions were done: Alkaline and acid phosphatase, alpha-naphthylacetate-esterase, naphthol-AS-acetate-esterase, 5' nucleotidase, ATPase, and acetylcholinesterase. The various enzyme reactions were sometimes done in combination and reticulum and collagenous fibers were investigated by a subsequent staining of argyrophilic fibers. The fine localization of the 5'-nucleotidase activity was studied ultrahistochemically. Adrenergic nerve fibers were investigated fluorescence microscopically using the glyoxylic acid method. PMID- 3051775 TI - The fourteenth International Conference on Yeast Genetics and Molecular Biology. 7-13 August 1988. Book of abstracts. PMID- 3051774 TI - [Principles of functional evolution at the cellular, organ and organism levels (exemplified by the kidney and water-salt homeostasis)]. PMID- 3051777 TI - The activities of the nutrition section: alternatives to intersectoral nutrition planning. PMID- 3051776 TI - Nutrition, health and education of women in Papua New Guinea. PMID- 3051778 TI - [Surgical diseases of the adrenal glands in childhood--pediatric aspects]. AB - Detailed discussion of diseases of the adrenals in children where surgery may be indicated, seen from the paediatric point of view. Following differentiation between adrenal insufficiency and adrenal hyperfunction, as well as adrenal haemorrhage--where differential diagnosis is often rather difficult--the tumours of the zona glomerulosa, fasciculata and reticularis as well as of the adrenal medulla are presented and their signs and symptoms, their clinical hormonal diagnosis, localisation diagnosis and therapy are described. PMID- 3051779 TI - [Indications for the surgical intervention in endocrine disorders of the ovaries]. AB - Endocrine disturbances of the ovaries with indication for surgery are rare. They require close cooperation between paediatric endocrinologist and paediatric surgeon. Clarification is mainly based on endocrinological findings and ultrasound examination of the ovaries. The following disturbances are discussed: Central (genuine and peripheral or pseudo-) precocious puberty, hormonally active ovarian tumours and cysts, the various forms of dysgenesis of the gonads, especially tumour-prone mixed gonadal dysgenesis with XO/XV karyotype, the syndromes of polycystic ovaries, and girls who grew up as boys with male external genitals. PMID- 3051780 TI - [ABH and Lewis antigens of tracheal glands. II. Lewis negative individuals]. AB - Immunocytochemical studies were performed on tracheal wall samples embedded in paraffin; the samples were taken at 23 autopsies. In all cases, the red cells had been typed in postmortem serological studies as being Le(a-b-). Blood-group antigens were demonstrated by the indirect immunoperoxidase technique, using monoclonal Anti-A, Anti-B, Anti-Lea and Anti-Leb; H was detected by UEA 1. The secretor characteristics could clearly be diagnosed from the ABH staining pattern of the mucous glands. In 11 cases, the lewis antigen labeling patterns were identical to the group of Lewis-positive individuals. It seems probable, from the statistical point of view, that these 11 individuals were, in fact, Lewis positive and that the negative serology resulted from deterioration of the cadaver blood samples. The immunocytochemistry was quite different in the remaining 12 cases: (a) secretors (n = 9) were completely negative for Lea, Leb was equally negative in one case, but in the remainder it was detectable within mucous epithelia in minimal amounts and in an atypical granular distribution; (b) nonsecretors (n = 3) reversely exhibited complete negativity for Leb but a minimal staining for Lea. These findings are in harmony with the well established Lewis serology typing of secretions in Lewis negative individuals. Thus, a minimal Lewis antigen biosynthesis and secretion seem to occur in the absence of the Le gene: A alpha-4-L-fucosyltransferase of low activity might be the product of the allele le. PMID- 3051781 TI - Detection of ABO blood group-active glycolipids extracted from red cell membrane and heat hematoma using TLC-immunostaining. AB - Glycolipids extracted from groups A, B, and O erythrocytes were developed on thin layer plates; their ABO blood group antigenicities were detected by immunostaining method using avidin-biotin-complex (ABC). Among series of glycolipids of different flow rates, antigen-specific staining was observed in five bands from group A1 erythrocytes, four bands from group B, and two bands from group O. Monoclonal anti-A, -B, and -H antibodies specifically stained glycolipids from A1, B, and O erythrocytes, respectively. ABO blood grouping was possible from 5 g of epidural heat hematoma of a charred body by this method. ABC immunostaining on thin-layer chromatography is a useful and reliable method for ABO blood grouping in forensic practice. PMID- 3051783 TI - [Significance of fluorides in chronic hemodialysis with reference to fluoridation of drinking water]. AB - The metabolism of fluorides in impaired renal function is described. The use of fluoridized drinking-water in the production of dialysis fluid is discussed controversial. In patients treated by chronic hemodialysis the plasma fluoride level rises at a content of 1 mg fluoride per liter dialysis fluid. A relevant effect on the state of health is not certain established today. Therefore, it will be advised to the use of fluoride-free dialysis water. Its production should be made by use of a reverse-osmosis unit. PMID- 3051782 TI - [The significance of collagen in determining the age of a wound]. AB - The aim of the present study was to find out whether the quantitative and/or qualitative analysis of collagen can be useful in the determination of the age of healing skin wounds in various body regions of patients of different ages and sex. The quantitative measurement of total wound collagen revealed no clear time dependent changes. Immunohistochemical staining for collagen types I and III demonstrated the presence of type III as early as 2 days after injury. Procollagen type I was found in wounds beginning from 4 days on, while type I collagen was not present before 6 days after the injury. Immunohistochemical analysis of specific basement membrane proteins, collagen type IV and laminin, showed a reconstruction of the epithelial basement membrane beginning from on day 5, while a completely rebuilt basement membrane was found in most cases after more than 14 days, depending on the dimension of the wound and its treatment. PMID- 3051784 TI - [Status and trends of curative therapeutic procedures in localized prostate cancer]. AB - The first 10-year-treatment results may allow an assessment of various therapeutic methods of localized prostatic carcinoma at which the question of survival rate and the spectrum of complications take as a basis. In localized prostatic carcinoma the individual character of curative therapeutic regimens has been increased. The new UICC classification come up to this development. The radical operation is the standard method in the treatment of curable cancer (T1 T2) with most superior survival rates. A safe intraoperative bladder-urethra anastomosis and the postoperatively possible urethrotomia interna may reduce urinary incontinence and stenosis of the anastomosis. The use of erectile protective PE-technique should be verified with regard to its radicality. The interstitial radiation therapy using J 125 seeds or Iridium 192 is of superior therapeutic value. Whether the perineal ultrasound-guided implantation of J 125 or Ir 192 are advantageous should be checked by further investigation. A benefit from this method is its safe of potency; it may be used in cancer with low grading and tumor stage, but a close urological-radiological co-operation, a special equipment and radiation protection measures are necessary. Radiation therapy is indicated in older patients with localized tumors (T1-T3), but further reduction of irradiation side-effects and a biological judgement of the tumor by posttherapeutic biopsies be required. Since the prognosis of the prostatic cancer patients is dependent on lymphogenous metastatic spread its determination should be demand prior to localized therapy. In radical prostatectomy the pelvic lymph node dissection is performed simultaneously.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3051785 TI - [Nitrogen regulation in microorganisms]. AB - The central role of NH4+-assimilation in the microbial metabolization of several inorganic nitrogen sources and the regulation of its key enzymes--glutamate dehydrogenases (GDH--EC 1.4.1.2. and EC 1.4.1.4.), glutamine synthetase (GS--EC 6.3.1.2.) and glutamate synthase (GOGAT--EC 1.4.1.3.)--are presented. In excess of ammonia gramnegative bacteria as well as yeasts assimilate this ion in a NADH + H+ or NADPH + H+ dependent reaction by GDH. Under NH4+-limitation--in natural environments rather the rule than the exception--the ammonia assimilation is ATP dependent and catalyzed via GS and GOGAT. Subsequently the connection between the nitrogen metabolization and the resulting changes in the extracellular pH of growing yeast cultures is discussed. The stoichiometric exchange between NH4+ and H+ led to the assumption that in the physiological pH-range an energy dependent NH4+/H+ transport is the preferred++ mechanism of ammonia uptake for NH4+ excess as well as for NH4+ shortage. PMID- 3051786 TI - [Oxygen regulation of nitrogen metabolism in microorganisms]. AB - Oxygen is one of the most important environmental factors for microorganisms. Many metabolic reactions of aerobic or facultative anaerobic bacteria are influenced by varying oxygen concentrations. A lot of enzyme reactions in respiration processes, catabolism, anabolism and gene expression depend upon oxygen. Other enzymes such as nitrogenase or hydrogenases can be inhibited by increasing oxygen levels. Also complex metabolic processes including anaerobic respiration and fermentations are regulated by oxygen. Finally toxic oxygen derivatives have to be eliminated by living cells to overcome damage of cell constituents. In this way also bacteria which are included into the nitrogen cycle in the nature are influenced by oxygen. The different strategies of microorganisms to protect their nitrogenases for oxygen inactivation and the regulation of dissimilative nitrate reduction by oxygen are demonstrated in detail. PMID- 3051787 TI - [Sigmoid diverticulitis]. AB - Colonic diverticulitis has become a relatively frequent disease, with the sigma being affected by up to 95 per cent of all cases. Its clinical relevance has proved to depend on the prognosis of life-threatening diverticulitis complications. Sixty patients who had received surgical treatment at the Surgical Department of Potsdam District Hospital were analysed. That analysis together with evaluation of literature has been used for therapeutic recommendations. For cases of sigmoid diverticulitis early elective intervention is advised. The method of Henri Hartmann is considered an effective approach to diffuse peritonitis. PMID- 3051788 TI - [Selective proximal vagotomy with and without pyloroplasty--results of a randomized clinical study of duodenal ulcer 5 and 8 years after surgery]. AB - Selective proximal vagotomy (SPV) with pyloroplasty (n = 39) and without (n = 39) were compared five and eight years after surgery in a randomized clinical study. Pyloroplasty according to Finney had no effect on ulcer recurrence rates after both periods of time (7.9:7.9 or 10.8:8.3 per cent). Additional pyloroplasty, too, had no statistically significant effect on incidence and severity of post vagotomy syndrome, dumping and diarrhoea, though some trend towards an increase was discernible in wake of pyloroplasty. Occurrence of the lactase deficit syndrome after SPV with pyloroplasty was increased with statistical significance. Lasting loss of senses of appetite and saturation was observed for the first time after SPV, on top of the lactase deficit syndrome. Clinical results recorded after five and eight years did not reveal any significant difference between both groups. These findings are likely to suggest that SPV without pyloroplasty is sufficient for surgical treatment of duodenal ulcer. However, pyloroplasty in combination with SPV has continued to be fully indicated for no-resection therapy of ulcerogenic pyloric stenosis. PMID- 3051789 TI - [Value of immediate operation in acute cholecystitis]. AB - Operations were performed on 221 patients for tentative diagnosis of acute cholecystitis at the Surgical Department of Goslar District Hospital, between January 1982 and August 1986. Evaluation was possible of 179 of these cases. This group was compared with 96 patients who had undergone surgery some time after admission to hospital, between January 1980 and December 1981, with 90 patients of these patients being evaluated. Mortality, postoperative complications, length of surgical sessions, and length of hospitalisation of immediate treated patients were lower than those recorded from patients with delayed operations. Mixed germ cultures were recordable from the former group and monocultures from the latter. On balance, the rate of complications following immediate surgery did not deviate from data given for early surgery in the literature, in that it was below the rate following interval operation. Therefore, immediate surgery in the authors' hospital is considered to be the appropriate approach to acute cholecystitis. PMID- 3051790 TI - [Possibilities for using emergency abdominal ultrasound study and its significance in acute diseases of the abdominal cavity]. AB - Studies were conducted by the authors' of this paper into the diagnostic value of ultrasonic examination to clear up suspicion of acute abdominal problems, with 381 examinations being actually evaluated. Definitely valid diagnosis was obtained from the method in 37.7 per cent of these cases. The tentative diagnosis was invalidated in 18.2 per cent. The results are compared with literature data. Continued systematic use of the approach is recommended. It would bring down the time required for diagnosis and might provide another possibility of supporting the diagnostic effort which is quite difficult, when it comes to acute abdominal problems. PMID- 3051791 TI - [Our surgical heritage. Early evidence of biliary surgery]. PMID- 3051792 TI - [Our surgical heritage. The Thuringian Society of Surgery. A short retrospect of 4 decades]. PMID- 3051794 TI - [Does the Baker anastomosis still have significance?]. PMID- 3051793 TI - [Abdominal pain in metabolic disorders. Guidelines for patient management for the general surgeon]. AB - Hyperglycaemic-ketotic disorder has proved to be the most common cause of pseudoperitonitis. Porphyria, hypercalcaemic crises, and Addisonian crisis may be accompanied by colicky abdominal pain, nausea, vomiting, and electrolyte shift. Poisoning, too, may be associated with acute abdominal symptoms. However, absence of objective signs, such as paralysis, or absence of both spontaneous rigidity of the abdominal wall and inflammation may be indicative of pseudoperitonitis. In routine laboratory checks attention should be given to unusual deviations. PMID- 3051795 TI - [Anastomoses of the large intestine in standardized diffuse peritonitis in the rat]. AB - A standardized experimental diffuse peritonitis model was developed for the purpose of studying the mechanisms of impaired anastomotic healing in cases of peritonitis. Sham peritonitis or standardized diffuse peritonitis was induced to 42 animals, and anastomosis of the ascending colon was performed 24 hours later. Bursting pressures were measured at six different time intervals, following performance of anastomosis. Significant differences in bursting pressure between sham peritonitis and real peritonitis were restricted to the shortest time interval after performance of anastomosis. More studies will be necessary to investigate the high rates of insufficiency in peritonitis cases. PMID- 3051796 TI - [The East German Twin Study l984/85. Report of selected aspects]. AB - Analysis of a multicentre study of the GDR in which 67 clinics with a total of 1,200 twin pairs participated over a scheduled period of 18 months. In the result a recommendation for the antenatal, intrapartal and neonatal care of gemini pregnancy was prepared. PMID- 3051797 TI - [Development and responsibilities of clinical obstetrics and gyne- cology in the 19th century at the Greifswald University--a contribu- tion on the 110 year existence of the clinic of gynecology and obstetrics]. AB - At the university of Greifswald clinical obstetrics developed at the beginning of the 19th century with priority for education of physicians and midwives. For further support of education in the middle of the century an obstetrical outpatients' department has been founded. Approximately at the same time gynaecological diseases were treated in both institutes so that the teaching combination obstetrics and gynaecology gradually came into existence and medical attendance and scientific work became more importance. There was a similar process at other German universities in the last century. PMID- 3051799 TI - [Diagnostic chorionic biopsies in the first trimester. Report of experiences over a 2 year period]. AB - We report about 58 ultrasonographically guided transcervical chorionic villus biopsies from January 1985 to November 1987. Maternal age greater than 35 years (n = 28), followed by trisomy 21 or 18 (n = 10) were the mean indications. Biochemically evaluation of storage diseases (n = 6) and genomically DNA-analysis because of phenylketonuria (n = 2) were combined in each case with cytogenetic diagnosis. In the other cases certain indications were the reasons for biopsy. In 52 of 58 cases we were successful in biopsies and diagnoses. In the other 6 biopsy specimen we didn't found chorionic villi. 3 abortions we observed up to day 3 after operation (n = 3) and after 6 weeks (n = 1). Pathological findings were 1 trisomy 16, 1,47,XYY-karyotype and 1 embryo with phenylketonuria. Another reason for termination of pregnancy was male karyotype in a Morbus Duchenne-risk and 1 risk for Rett-syndrome. Meanwhile 30 healthy babies were born. PMID- 3051798 TI - [Results of invasive and pharmacosonographic studies in pregnancy (I)]. AB - In prenatal diagnosis besides radiographic methods there is a growing trend towards the use of invasive diagnostic techniques. These techniques have proved to be suitable for early detection of genetically determined conditions, diseases, and malformations of different origin. Invasive examination techniques were applied to 1.104 cases of prenatal diagnosis at Humboldt-University, Berlin, School of Medicine (Charite), Department of Gynaecology and Obstetrics, from January 1st, 1985, up to September 30th, 1987. In this paper importance and possible applications of these methods have been reported. PMID- 3051800 TI - Anemia and anti-erythrocyte antibodies developed after repeated injections of sonicated preparations of Plasmodium berghei and Babesia rodhaini. AB - Anemia developed in mice after repeated injections of sonicated preparations of Plasmodium berghei and Babesia rodhaini. Anti-erythrocyte antibodies were detected in sera by an enzyme-linked immunosorbent assay and their levels rose as anemia developed. Administration of carbon particle suspension to block the mononuclear phagocyte system was effective to inhibit anemia induced by injections of sonicated parasite preparations. Sodium dodecyl sulfate polyacrylamide gel electrophoresis and immunoblotting analyses were carried out to examine autoantigens on the surface of the erythrocyte membrane reacting to anti-erythrocyte antibodies. The anti-erythrocyte antibodies from mice injected with the sonicated P. berghei preparations bound specifically to three components of the erythrocyte membrane. The anti-erythrocyte antibodies produced by the sonicated B. rodhaini preparations also reacted with four components, two of which corresponded to those reacting after the injection of the sonicated P. berghei preparations. These results suggest that both P. berghei and B. rodhaini organisms have some materials capable of inducing production of the anti erythrocyte antibodies which mediate erythrophagocytosis by macrophages of the mononuclear phagocyte system during Plasmodium and Babesia infections. PMID- 3051801 TI - Toxins and serotypes of faecal non-enterotoxigenic and non-enteropathogenic Escherichia coli strains causing mannose-resistant haemagglutination: relation with haemagglutination patterns. AB - Forty-three faecal non-enterotoxigenic and non-enteropathogenic human Escherichia coli strains causing mannose-resistant haemagglutination (MRHA) were tested for production of cytotoxic necrotizing factor (CNF), haemolysis (Hly), Verotoxin (VT) and lethal activity for mice. The serotypes of the strains were also determined. Of the total strains investigated, 49% synthesized CNF, 53% were haemolytic and 40% were lethal for mice. No strain producing VT was detected. Striking differences in the production of Hly and CNF were observed when MRHA strains were grouped according to their lethal or non-lethal activity. Thus, 82% of lethal strains produced Hly and/or CNF whereas only 35% (p less than 0.01) and 27% (p less than 0.01) of non-lethal strains produced Hly and CNF, respectively. The production of toxins was specially associated with strains possessing defined MRHA types. Thus, 100%, 82% and 50% of strains belonging to MRHA types III, IVa and V, respectively, were toxigenic, whereas no toxigenic strains from MRHA types IVb and VI were detected. The majority (77%) of MRHA strains possessed typical O groups usually reported to be present in pathogenic extraintestinal E. coli or in facultatively enteropathogenic E. coli. Furthermore, these O groups were more frequently detected in toxigenic (93%) than in non-toxigenic (47%) strains (p less than 0.01). Our results suggest that faecal non-enterotoxigenic E. coli strains belonging to MRHA types III, IVa and V may be responsible for extraintestinal infections as well as for sporadic intestinal infections, and that certain O groups are specially associated with E. coli strains belonging to particular MRHA types. PMID- 3051802 TI - A comparative study of the effect of oral treatment with augmentin, amoxycillin and bacampicillin on the faecal flora in mice. AB - Oral treatment of groups of four mice with different daily dosages of three related antibiotics, amoxycillin, augmentin and bacampicillin, has indicated the influence of the amount of the dose that reaches the intestine in a biologically active form. Augmentin (amoxycillin plus clavulanic acid to protect it against enzymatic hydrolysis) appeared to have a suppressive effect on the indigenous colonization-resistance-associated microflora. Dose-effect curves of amoxycillin alone, showed the same shape but at a lower level. Bacampicillin treatment practically did not have an effect on the faecal flora. Only at doses of bacampicillin of well above 1.5 mg per day, an indication was seen of CR-flora disturbance. At a dose level of 2 mg and more per day, a low concentration of beta-aspartylglycine was found in the faeces. A normal undisturbed intestinal flora normally produces in mice sufficient enzyme to degrade completely this dipeptide released by the host organism into the intestinal tract. PMID- 3051803 TI - Electron microscopic study of an experimental combined infection of just-weaned piglets with transmissible gastroenteritis virus and K88ac positive enterotoxigenic Escherichia coli. PMID- 3051804 TI - The construction, selection, characterization, and application of recombinant herpes viruses. PMID- 3051805 TI - [Problems of neurobiology and the development of memory]. PMID- 3051806 TI - [Tecto-thalamo-telencephalic visual system of the brain in 13-day-old chick embryos]. AB - Light and electron microscopic studies have been made of the nervous tissue in three parts of the tecto-thalamo-telencephalic visual system--i.e. tectum opticum, nucleus rotundus of thalamus and ectostriatum of telencephalon--of 13 day chick embryos. Neuroblasts and neurones at various stages of differentiation were described together with various types of synaptic and nonsynaptic intercellular contacts in the neuropil of these brain structures. Heterochronous maturation of these parts of the visual system in embryogenesis was noted which reflects the level of their phylogenetic maturity. Being phylogenetically more ancient structures, tectum opticum and nucleus rotundus reveal differentiation earlier than ectostriatum which is phylogenetically younger. PMID- 3051807 TI - [Characteristics of household and occupational contacts of the population with natural foci of tick-borne encephalitis in the past and today]. PMID- 3051808 TI - [Differentiation of virulent strains of Shigella and entero-invasive Escherichia from their avirulent variants using modified indirect immunoenzyme analysis]. AB - The data obtained in this investigation confirm that the modified indirect enzyme immunoassay (EIA) permits the differentiation of virulent bacteria of the genus Shigella and enteroinvasive Escherichia (group 1), regularly containing virulence plasmids with a molecular weight of 120-140 MD, from their avirulent variants which have lost these plasmids (group 2). The ratio of the optic density (OD) values of the positive control samples (the OD of group 1) to the OD values of the negative ones (the OD of group 2) is significantly higher than 2.1. As revealed by EIA, the differences between groups 1 and 3 (avirulent Shigella strains and E. coli smooth strain O124, retaining high-molecular plasmids with a molecular weight of 120-140 MD or their fragments) are statistically insignificant. The ratio of the OD of group 1 to the OD of group 3 is significantly less than 2.0. Analysis of outer membrane protein (OMP) preparations isolated from S. flexneri virulent strain 2a and its isogenic avirulent plasmid-containing variant has revealed significant differences in their EIA results. The ratio of the OP of OMP preparations isolated from the virulent strain to the OD of OMP preparations from the avirulent strain exceeds 2.1. PMID- 3051809 TI - [The problem of nutrient media today]. PMID- 3051810 TI - [The boundaries of the family Enterobacteriaceae]. PMID- 3051811 TI - Associations between a fluorescent DNA ligand, 4',6-diamidine-2-phenylindole.2HCl (DAPI), and RNA. AB - The experiments performed in vitro have shown that DAPI and RNA form insoluble and indigestible complexes. This seems to explain the earlier observed retardation of drug accumulation in the nucleus of a living cell in the presence of RNA. PMID- 3051812 TI - Changes in pH and acid-base equilibrium during experimental liver transplantation by active and passive veno-venous bypass. AB - The effect of experimental liver transplantation on pH and the acid-base equilibrium by active and passive veno-venous bypass was examined. It is concluded that the use of active veno-venous bypass may considerably decrease but not completely prevent the development of metabolic acidosis by elimination of the phase. PMID- 3051813 TI - The importance of closed bladder irrigation in prostatectomy. AB - Closed bladder irrigation in transvesical prostatectomy is an important factor in arresting bleeding and preventing or diminishing bacterial infection. A bladder irrigation of appropriate intensity may ensure that the blood clots be removed rapidly from the bladder and thereby the formation of larger clots and bladder tamponade can be avoided. The bacteriological examination of the eluents of 82 patients revealed that the longer the patient is maintained on a catheter, the longer bacteriuria and pyuria will persist. Two days are considered by the authors to be the optimal time of maintaining a closed bladder irrigation. PMID- 3051814 TI - Transvesical and transabdominal closure of vesicovaginal fistulas. AB - Between 1973 and 1985, a total of 46 patients were operated for vesicovaginal fistula. Depending on the extension and localization of the fistula, transvesical or transperitoneal closure was performed. In 12 patients ureteral neoimplantation was made. Fistula recurrence was observed in 6 cases. Based on their favourable experience, authors recommend this surgical solution for the closure of fistulas adjacent to the ureteral orifices and/or for that of larger fistulas. PMID- 3051815 TI - Pathological consequences of the vanishing twin. AB - Fetus papyraceus is a mummified, compressed fetus occurring in association with a viable twin. The death of the fetus usually occurs early in the second trimester. A co-twin dying earlier may be absorbed completely, whilst later fetal death usually results in macerated, but not compressed fetuses. This course of events can be well demonstrated by ultrasonography. The death of one fetuses may be associated with minor malformations of the surviving one. After termination of twin pregnancies the detailed check-up of the newborn and histopathological examination of the placenta is essential. PMID- 3051816 TI - In memoriam Andre Cournand (1895-1988). His role in the development of modern cardiology. PMID- 3051817 TI - Myocardial ischemia in 1988. What is it? PMID- 3051819 TI - [Role of external biliary drainage in common bile duct lithiasis. Apropos of 172 case reports]. AB - A series of 172 lithiasis of the common bile duct has been analysed. Priority has always been given to the external drainage by a T tube. It seems to be the simplest method, with the lowest morbidity. 6 patients, as 3.48% are deceased precociously after the intervention in keeping with the gravity of the lesions and the general state. PMID- 3051818 TI - [Treatment of extrahepatic bile duct atresia: Kasai's operation or hepatic transplantation?]. AB - The authors analyse the records of 60 children who had a failed Kasai procedure and were referred for liver transplantation between 1984 and 1986. By the end of 1987, 45 had been transplanted, eight had died before transplantation, four had been excluded while three were still waiting for a potential donor. Actuarial survival at 2 and 3 years of the 30 children with a follow-up greater than one year is 79 +/- 8%. Nowadays, the surgical therapy of biliary atresia should include liver replacement. The original porto-enterostomy (one Roux-en-Y loop, no stoma) according to Kasai should remain the first surgical procedure performed under 8 weeks by a trained surgeon. Good long-term results can be expected in 30 to 40%. In case of straight failure, liver replacement should be recommended in early age and, in case of delayed failure, before the age of 6 to avoid chronic disabling. PMID- 3051821 TI - Evaluation of the effects of elastic compression in patients with chronic venous hypertension by laser-Doppler flowmetry. AB - The evaluation of the effects of elastic compression in patients with venous hypertension and perimalleolar ulceration may be performed using laser-Doppler flowmetry (LDF). This technique may reveal microcirculatory positive variations which are not evident by standard methods as Doppler, plethysmography and ambulatory venous pressure (AVP). In this study we demonstrated the good correlation between AVP values and laser-Doppler parameters, particularly the resting flow and the venous vasomotor response. Also the response of the skin of the perimalleolar region to a local increase of skin temperature was well correlated with AVP parameters. By evaluation of laser-Doppler parameters it was possible to differentiate normals from abnormals (patients with venous hypertension) and it was possible to show variations of microcirculation associated with the decrease of the areas of ulceration after 3 week elastic compression. No significant variations have been recorded by AVP before and after 3 weeks of treatment with elastic compression in 80 patients with venous ulcerations. Laser-Doppler flowmetry was useful to evaluate the positive changes produced by elastic compression. PMID- 3051820 TI - Correlation of duplex scanning and two plane angiography in patients with symptomatic carotid stenosis. AB - The correlation between flow velocity and percent stenosis in carotid arteries was assessed in 43 symptomatic patients (86 carotids) using duplex scanning and angiography. Duplex scanning evaluation was performed using the ATL Ultramark 4 duplex scanner with the 5-7.5-10 MHz probe. Using the ratio of peak velocity in the internal carotid in systole (VICA) to the end diastolic velocity in the common carotid (VCCA) we studied its correlation with the percent stenosis observed by angiography. A good correlation (r = .95) was found. The graph obtained indicates that a significant (greater than 50%) stenosis may be graded by the VICA/VCCA ratio with a high degree of accuracy (97%). This may be useful not only for screening patients to select for angiography and surgery but also to follow up the natural history of carotid stenosis. PMID- 3051822 TI - [Clinical approach to the patient with polyvascular disease]. AB - The history and the physical examination are important steps in the clinical approach to the polyvascular patient. They are the foundation of a rational diagnostic and therapeutic management. Complementary investigations are nevertheless necessary. Ultrasonography has become an essential complement to the physical examination. Its field covers all peripheral vessels: supra-aortic arteries, aorta and its branches, limb arteries. The ultrasound are applied following continuous and pulsed Doppler and real time echography. The two methods may be combined (duplex echography). The investigations are performed first at rest, but assessments under strain are often necessary. The cardiac examination has been enriched by modern methods: thallium scintigraphy under dipyridamole is specially useful in order to measure the coronary reserve. Angiographic images are still a must for most of the aggressive therapeutic decisions. The modern techniques are more flexible using digitalization and a suitable investigation is chosen in function of each individual condition. PMID- 3051823 TI - Results of subxiphoid pericardiostomy in pericardial effusion. AB - 37 patients with pericardial effusions were managed by early subxiphoid pericardiostomy under local anaesthesia in most cases (31/37). The aetiology was pyogenic infection in 21, tuberculosis in 7, trauma in 3, cardiomyopathy in one. It was unclear in five patients. The effusion recurred in two patients (one with pyogenic, the other with traumatic pericarditis). An infant with pyogenic pericarditis developed pericardial constriction. The mortality, mainly due to associated infections or diseases, was related to aetiology: 0% for tuberculosis and trauma, 19% (4/21) for pyogenic infections, 80% (4/5) for unclear causes. Overall results were good in 28 patients. Subxiphoid pericardiostomy is a simple and safe procedure which allows a quick differential diagnosis and drainage of pericardial effusions. PMID- 3051825 TI - Non-invasive functional investigation of chronic venous insufficiency with special reference to foot volumetry. PMID- 3051824 TI - Investigations on the pathogenesis of venous leg ulcers. AB - Based on a prospective study of 92 patients with DVT initiated in 1979, including a follow-up every year, the following investigations were performed: phlebography, Doppler-ultrasound, plethysmography (strain gauge and PPG) and foot volumetry. In ulcer-patients skin blood flow was measured by Laser-Doppler and local oxygen supply by measurement of transcutaneous oxygen. Transcapillary protein-leakage was assessed by isotopic methods, local lymph-drainage by isotopic lymphography and indirect x-ray lymphography. From the results of these investigations the following course of events, which may be of importance for the development of venous ulceration, can be outlined: 1. Venous refluxes penetrating into the venules of the skin. 2. Chronic venous ambulatory hypertension (lack of pressure-fall during walking), waves of high pressure in the capillaries. 3. Protein-rich oedema. 4. Failure of fibrin clearance from the pericapillary space due to inadequate (exhausted?) fibrinolysis and deficient local lymph drainage. 5. Resulting changes in the ground substance, proliferation of fibres. 6. Local hypoxia due to a diffusion block by the impermeable pericapillary cuffs, partially due to a reduction of capillaries in the superficial layers. PMID- 3051826 TI - Iodosorb compared to standard treatment in chronic venous leg ulcers--a multicenter study. PMID- 3051827 TI - Surgical management of chronic venous insufficiency. AB - Chronic venous insufficiency is a pathologic condition of the skin and subcutaneous tissues in the lower extremity caused by stasis of the blood flow. Incompetency or failure of the venous valves results in reflux and ambulatory venous hypertension, which is more severe with deep than with superficial venous incompetency. Superficial chronic venous insufficiency (varicose veins) is effectively managed with ligation and stripping of incompetent perforator and superficial veins to restore normal venous physiology. Deep chronic venous insufficiency (postphlebitic leg) presents a widespread pathologic disorder that is refractory to surgical correction. Adjunctive surgical measures such as removal of incompetent perforators or superficial veins to lessen local stasis or skin grafting of ulcers are often indicated in selected cases. The underlying chronic venous insufficiency requires management with elastic compression, elevation of the legs, and exercise for best results. PMID- 3051828 TI - The bimodal effect of interleukin 1 on rat pancreatic beta-cells--stimulation followed by inhibition--depends upon dose, duration of exposure, and ambient glucose concentration. AB - To investigate the hypothesis that interleukin 1 initially stimulates and then suppresses beta-cell function and that this sequential effect is directly related to interleukin 1 dose, duration of exposure, and ambient glucose concentration, insulin release was measured from cultured newborn rat islets exposed for 6 h to 6 days to interleukin 1 at doses ranging from 20 to 2000 ng/l at glucose concentrations of 3.3, 5.5 and 11 mmol/l. After 6 h of exposure and at all three glucose levels, all doses of interleukin 1 stimulated insulin release, maximal stimulation (370% of control) being observed at 5.5 mmol/l glucose and 100 ng/l interleukin 1. In contrast, after 6 days, all doses of interleukin 1 were inhibitory irrespective of glucose level, maximal inhibition (90%) being observed at 11 mmol/l glucose and 2000 ng/l interleukin 1. At 24 and 48 h of exposure, the biphasic effect of interleukin 1 was observed: lower doses of interleukin 1 at lower glucose concentrations at 24 h being more stimulatory with transition to inhibition directly related to higher glucose levels, higher interleukin 1 doses, and longer exposure. After 48 h, 200 ng/l of interleukin 1 increased insulin release to 220% at 3.3 mmol/l glucose, but at 11 mmol/l glucose a 60% suppression was seen. On the basis of these data we suggest that interleukin 1's effect on beta-cells is bimodal: stimulation followed by inhibition. Increasing interleukin 1 dose and ambient glucose concentration shift this response to the left. Experimental results will, and in vivo effects may, depend upon these three variables. PMID- 3051829 TI - The significance of plasma membrane transport of iodothyronines in the regulation of thyroid hormone bioavailability. PMID- 3051830 TI - Biochemistry of iodothyronine deiodination. AB - Extrathyroidal deiodination of the thyroidal main secretory product L-T4, which may have prohormone functions, reveals hormonally active and potentially regulatory potent triiodothyronines. The regulation of these enzyme reactions is still unknown but three deiodinase isoenzymes can be classified based on their physicochemical, kinetic, pharmacological and physiological properties. The purification to homogeneity and cloning of the substrate binding 27kDa subunit of 5'-deiodinase type I is currently performed and these experiments suggest a close similarity of this subunit in 5'D I and II, no evidence is found yet supporting a structural relationship to other ITH binding, transport, metabolizing or receptor proteins, in spite of a high similarity of the hormone binding sites of 5'D I and hTBPA. PMID- 3051831 TI - [Ether bond cleavage of thyroxine to diiodotyrosine (DIT). DIT in serum, a possible new marker of leukocyte activity in sepsis and severe infections]. PMID- 3051832 TI - Is there a physiological role for reverse triiodothyronine? PMID- 3051833 TI - Hepatic metabolism, biliary clearance and enterohepatic circulation of thyroid hormone. PMID- 3051834 TI - Systemic adrenergic actions on liver T-4 5' deiodinase differ considerably from those seen in vitro. PMID- 3051835 TI - Causes and effects of the low T3 syndrome during caloric deprivation and non thyroidal illness: an overview. AB - The increased serum reverse T3 and decreased T3 during caloric deprivation and non-thyroidal illness is caused by decreased T3 production (with intact degradation) and reversed T3 degradation (with intact production) respectively. These changes can ensue from two mechanisms i.e. decreased 5'D of T4 and of reverse T3 (possibly caused by a decrease in naturally occurring reducing agents) or by decreased transport of T4 and reverse T3 into the liver (possibly caused by decreased ATP concentrations in the liver). The effects of the low T3 syndrome at the tissue level are in many instances comparable to those seen in hypothyroidism. The effects lead to conservation of energy and decrease of protein breakdown. These effects are considered to constitute a beneficial adaptative mechanism in situations in which the organism is endangered. There is no evidence that treatment of patients with the low T3 syndrome with thyroid hormones is of any benefit. Knowledge at the present moment suggests that administration of thyroid hormones during caloric deprivation or non-thyroidal illness should be avoided. PMID- 3051836 TI - Syndromes of thyroid hormone resistance. PMID- 3051837 TI - Thyroid hormone metabolism in the central nervous system. PMID- 3051838 TI - Group reminiscence therapy as a nursing intervention: an experimental study. Part one. PMID- 3051839 TI - Double-blind comparison of transdermal scopolamine, droperidol and placebo against postoperative nausea and vomiting. AB - Since transdermal scopolamine (TS) seems effective against seasickness, we compared its antiemetic effect with intravenous droperidol (DHBP), our routine antidote for postoperative emesis. Ninety-six female patients (ASA I-II) scheduled for short-stay surgery were randomly allocated to three study groups after giving their informed consent. The three groups were as follows: TS adhesive, delivering 140 micrograms initially and 5 micrograms/h thereafter + placebo 0.5 ml i.v. 5 min before the end of surgery; transdermal placebo adhesive preoperatively + DHBP 0.5 ml (1.25 mg) i.v. 5 min before the end of surgery; transdermal placebo + 0.5 ml placebo i.v. as indicated above. Oxycodone i.m. and glycopyrrolate i.v. were given for premedication together with the test adhesive. Anaesthesia was induced with thiopental and maintained with nitrous oxide and oxygen, enflurane, vecuronium and fentanyl. Neostigmine and glycopyrrolate were administered for reversal. In the recovery room no differences in nausea or vomiting were observed between the groups. Sedation was significantly more marked (P less than 0.15-0.0001) after DHBP than after either TS or the given DHBP and 6% of those given the placebo (P less than 0.05). During the following 24 h nausea was reported more by the placebo patients (25) than by those on TS (20) or DHBP (15) (P less than 0.05). However, actual vomiting on the ward did not differ between the groups. Visual disturbances were more frequent after TS (P less than 0.01). We conclude that prophylactic transdermal scopolamine does not diminish postoperative emetic sequelae. PMID- 3051840 TI - Brain resuscitation after cardiopulmonary arrest. PMID- 3051841 TI - Anesthesia and myocardial ischemia. PMID- 3051842 TI - Closed chest cardiopulmonary resuscitation: a review. PMID- 3051843 TI - Emergency airway management. PMID- 3051844 TI - Disadvantages of isoflurane in coronary artery disease. PMID- 3051845 TI - Vasopressors and local anesthetics. PMID- 3051847 TI - Inspiratory work of breathing during weaning from mechanical ventilation. PMID- 3051846 TI - Circulatory effects of epidural anesthesia in patients with cardiac disease. PMID- 3051848 TI - The complex problem of ARDS: pathophysiology, management. PMID- 3051849 TI - Ventilator-induced lung damage. PMID- 3051850 TI - Adverse effects of analgesics and intravenous anesthetic agents. PMID- 3051851 TI - Effect of supernatants of dermal leprosy granulomas on lymphocyte morphology and function. AB - A comparison has been made of the characteristics of dermal granulomas of tuberculoid and lepromatous leprosy by culturing them in vitro. The granulomas were derived from lesions of untreated patients and their effect was assessed on the morphology and function of lymphocytes derived from peripheral blood of normal individuals. The concentration of proteins released in the supernatants was similar in both the type of granulomas. However, the supernatants from the lepromatous granulomas markedly diminished the viability of lymphocytes when compared with supernatants derived from the tuberculoid granulomas. The supernatants from both the tuberculoid and lepromatous granulomas, contained soluble factors which depressed the 14C-leucine and 3H-thymidine incorporation by lymphocytes. The depression in 3H-thymidine uptake was more pronounced with the supernatants from the lepromatous granulomas while the diminution of 14C-leucine incorporation was more marked with supernatants from the tuberculoid granulomas. The supernatants did not show any migratory inhibitory activity in vitro. When the cells from the granulomas were dispersed and cultured in vitro, only very low concentration of proteins was detectable. PMID- 3051852 TI - CD1 positive epidermal Langerhans cells in indeterminate leprosy. AB - Langerhans cells (LC) in the skin lesions of 10 untreated indeterminate leprosy patients were defined by indirect immunofluorescence using monoclonal antibodies. No difference was observed in the numbers and distribution of epidermal LC in the indeterminate lesions and controls. However, the infiltrates of these lesions contained CD1+ cells. Most cells in the infiltrates HLA DR antigens. PMID- 3051853 TI - [Supervised bimonthly quadruple chemotherapy in 72 paucibacillary leprosy patients in French Guyana]. AB - In French Guyana we have treated 72 paucibacillary leprosy with an association of Rifadine + Lamprene + Trecator + Disulone given twice monthly under supervision during 6 months. Results have been satisfying and side effects rare. The sequential character of treatment shows a substantial advantage on the operational side but may appear to be favorable to bacterial resistance. PMID- 3051854 TI - Leprosy as a zoonosis: an update. AB - Naturally-acquired leprosy has been reported in nine-banded armadillos captured in the southern United States, a chimpanzee from Sierra Leone, and in two "sooty" mangabey monkeys from Nigeria. A significant prevalence of leprosy in wild armadillos establishes this animal as a reservoir of M. leprae, and exposure to armadillos has been implicated as a source of leprosy in humans. Current evidence suggests that leprosy is a zoonosis in certain nonhuman primate species. Control and eradication programs for leprosy should take into consideration the possible influence of extra-human sources of M. leprae, especially zoonotic leprosy. PMID- 3051855 TI - European atrial fibrillation trial. PMID- 3051856 TI - Thymolipoma in patients with myasthenia gravis: report of two cases and review. AB - Two cases of simultaneous occurrence of myasthenia gravis (MG) and thymolipoma are described and 4 previously reported cases are reviewed. In all 6 cases, thymectomy was performed. Pre- and postoperatively, the clinical status of the patients was similar to that of late-onset MG without thymolipoma. It is possible that simultaneous occurrence of MG and thymolipoma may be coincidental. PMID- 3051857 TI - Increased level of cerebrospinal fluid beta 2-microglobulin is related to neurologic impairment in multiple sclerosis. AB - beta 2-microglobulin was measured in the CSF of 33 MS patients (age range 26-66 years) and compared with the 95% confidence interval determined in an age-matched reference group of 32 patients without neurologic disease. Six MS (18%) had moderately increased concentrations and this subgroup was characterized by severe neurologic impairment according to Kurtzke's expanded disability scale. Increased CSF beta 2-microglobulin concentration in MS may reflect lesion formation. PMID- 3051858 TI - Fetal growth in the offspring of epileptic women: results of an Italian multicentric cohort study. AB - An historical cohort study of the association between maternal epilepsy, anticonvulsant drugs and fetal growth was carried out among 47 hospitals collaborating in the Italian Multicentric Registry on Birth Defects (IPIMC). Birth weight, head circumference and body length were studied in 164 babies of epileptic women and compared to 185 controls. Seventy-nine epileptic women were treated by monotherapy, 59 by polytherapy and 26 took no anticonvulsant during pregnancy. An intrauterine growth retardation and a smaller head circumference was observed among babies of epileptic women. No effect was evident for body length. When specific anticonvulsant therapies were taken into account, only phenobarbital showed an effect on birth weight and head circumference; a reduction in head circumference was observed also in the babies of untreated epileptic women. The other antiepileptics (carbamazepine and valproic acid) showed no influence on the outcome considered. The observed effects on fetal growth can be interpreted as a result of an interaction between the effect of the maternal disease and that of the anticonvulsants. PMID- 3051860 TI - Improving functional rehabilitation outcome following epilepsy surgery. AB - This article reviews the literature on rehabilitation outcome following epilepsy surgery to provide perspective on the research issues in examining vocational and independent living outcome. The existing literature does not suggest dramatic independent living or employment gains as a result of this surgery. Those most likely to profit in these areas are adult seizure patients with excellent surgical outcome, freedom from pre-existing impairments (psychiatric, neuropsychological, or financial dependence on subsidy), and recent presurgical vocational activity. Recommendations are offered toward improving this outcome. PMID- 3051859 TI - Surgery of epilepsy: methods. AB - The various surgical methods used in the management of epilepsy are reviewed. Intracranial recording procedures such as epidural and subdural recording are presented as well as their main indications, advantages and inconveniences. Emphasis is placed on the techniques of intracranial recording used at the Montreal Neurological Institute, namely chronic cortical and depth recording, both based on stereotactic localization procedures using MRI and DSA. The various resective procedures for the surgery of epilepsy are discussed, namely cortical resections in various locations such as frontal, central parietal, occipital and temporal areas. Special emphasis is placed on techniques of cortical resection in the dominant hemisphere, using local anaesthesia and topographic mapping. The indications and techniques of such procedures as callosotomy, selective amygdalohippocampectomy, hemispherectomy, and stereotactic interventions are also presented. PMID- 3051861 TI - Economic costs of epilepsy--treatment benefits. PMID- 3051862 TI - Consequences of severe epilepsy: neuropathological aspects. PMID- 3051863 TI - Consequences of severe epilepsy: psychosocial aspects. AB - Patients with severe epilepsy very often present a diversity of problems which interact with each other and with factors in the environment in subtle ways. Intellectual and social shortcomings as well as anxiety-related emotional problems are among the most common primary and secondary consequences seen in this patient group. Anxiety among the relatives, ignorance and prejudice in the general population often add to the patients' burdens. When seen from a developmental neuropsychological and psychological point of view the importance of early treatment must be stressed. PMID- 3051864 TI - Severe epilepsy: diagnostic and epidemiological aspects. AB - Epidemiological parameters of epilepsy have been estimated in a large number of studies. Reported annual incidence rates for recurrent seizures vary between 30 and 50/100,000, and prevalence rates in most studies between 500 and 1000/100,000. Varying findings are mostly due to different case ascertainment methods and definitions of epilepsy applied in different surveys. These aspects will be discussed in this paper. A special emphasis is laid on differential diagnosis of seizure disorders, and on the prevalence and causes of complex partial epilepsy. This epileptic disorder is one of the most common forms of epilepsy, and also difficult to treat in many cases. Many recent reports show that prognosis of seizures in patients with epilepsy is better than suggested in earlier studies. However, 25-30% of epileptics seem to have frequent (greater than 1/month) seizures. Our own study, in accordance with earlier findings suggests that prevalence of patients with severe complex partial epilepsy is about 80-90/100,000. Available literature provides several predictive factors for the prognosis of seizures and assessment of prognosis is possible quite early after the onset of seizures. Treatment choices for patients with intractable epilepsy will be discussed. PMID- 3051865 TI - The accuracy of the dichotic, the visual half-field, and the intracarotid sodium amytal memory tests in preoperative neuropsychological investigation of epileptic patients. AB - Three methods for determining the lateralization of memory functions in patients with pharmacologically resistant intractable partial epilepsy were compared and evaluated. The three methods studied were a dichotic memory test, a visual half field memory test, and an intracarotid Sodium Amytal memory test. A total of 35 epileptic patients considered for surgical therapy and a group of 20 non epileptic control subjects took part in the study. The results show that the three tests tap different memory functions and that each of the test pick up a memory dysfunction in the hemisphere indicated by EEG recordings. In combination with an extensive neuropsychological test battery, the three methods produce data that concur with the evaluation made of EEG recordings. PMID- 3051867 TI - In vitro interaction of M. leprae-infected Schwann cells and splenic cells. AB - The interaction between M. leprae-infected cultured Schwann cells and sensitized splenic cells was noted both under light and electron microscopy. No evidence of cytomorphological changes in infected Schwann cells was obtained. However, sensitized splenic cells were noted to undergo degenerative changes suggestive of the phenomenon of apoptosis. Subsequently a large number of these degenerated cells were observed within the Schwann cell. Such a process has not been hitherto reported in the histopathology of leprous nerves. Nevertheless, these findings indicate an aberrant metabolic function in M. leprae-infected Schwann cells. PMID- 3051866 TI - The role of neuroimaging in the surgical treatment of epilepsy. AB - Resective surgical treatment of medically intractable epilepsy requires accurate identification of the site and extent of the epileptogenic zone responsible for habitual seizures. Epileptogenicity per se is demonstrated electrophysiologically, but interictal and ictal EEG transients, whether recorded extracranially or intracranially, propagate widely and can give rise to false lateralizing and false localizing information. Neuroimaging techniques provide additional important information which greatly enhances confidence in localization derived electrophysiologically. Structural imaging with X-ray computed tomography and magnetic resonance imaging, as well as functional imaging with positron emission tomography, single photon emission computed tomography, and computerized mapping of electromagnetic activity, used together with other tests of focal functional deficit, 1) increase the confidence with which surgical resection can be performed on the basis of noninvasive tests alone, 2) aid in developing appropriate strategy for intracranial electrode recording when this is necessary, and 3) supplement results of invasive studies sufficiently to justify surgical resection in some patients who otherwise might be rejected for surgery. Addition of these new techniques, therefore, has increased the number of patients considered candidates for surgery, decreased the number of invasive procedures necessary before surgery can be performed, and increased the accuracy of surgical resection. PMID- 3051868 TI - Creutzfeldt-Jakob disease with intranuclear vacuolar inclusions: a biopsy case of negative light microscopic findings and successful animal transmission. AB - In a 53-year-old man with a progressive mental deterioration and myoclonic jerks, brain biopsy failed to show any significant light microscopical findings. Electron microscopy revealed membrane-bound vacuolar inclusions in many neuronal nuclei as the only prominent finding. Hamsters intracerebrally inoculated with the biopsy material demonstrated typical spongiform changes in the gray structures of the brain when sacrificed on the 309th and 332nd days post inoculation, characteristic of experimental Creutzfeldt-Jakob disease (CJD). These intranuclear vacuolar inclusions, originally reported in experimental Creutzfeldt-Jakob disease in this laboratory, may be a valuable electron microscopic feature in some CJD cases and may play an important role in supporting the diagnosis of CJD. PMID- 3051870 TI - Diagnostic accuracy of outpatient endocervical curettage using conventional and Vabra curettage of the cervix. AB - With the object of examining the diagnostic accuracy of and pain reaction to endocervical curettage (ECC) either by using a new instrument, Vabra mc cervix, or with a conventional 3 mm metal curette, 298 patients with histologically verified cervical intraepithelial neoplasia (CIN) or cytological suspicion have been involved in a consecutive prospective randomized trial. One hundred and forty-eight patients underwent curettage with the Vabra instrument (vabrasio) followed by conventional curettage; 150 underwent the same procedure, but in reverse order. In 114 patients, CIN was ascertained in one or both trials. Vabra mc cervix found normal histology in 23 of these patients (20%), whereas conventional curettage showed normal histology in 58 patients (51%). Quantitatively, Vabra mc cervix extracted significantly more tissue material than did conventional curettage. The pain intensity when using of Vabra mc cervix was less severe. The present study showed diagnostic accuracy of Vabra mc cervix to be 80% (95% confidence limits: 63.38-80.73), compared with 49% (95% confidence limits: 35.17-54.35) when using conventional curettage. PMID- 3051869 TI - Preoperative diagnosis of ovarian malignancies with intravenous digital subtraction angiography. AB - Intravenous digital subtraction angiography was performed on 59 patients with ovarian tumors. Dilatation of the uterine artery and its ovarian branch was relatively frequent in both benign and malignant tumor groups, while dilation of the ovarian artery, and hypervascularity from the uterine artery or the ovarian artery were fairly specific to the malignant tumor group. These five abnormal vascular patterns were evaluated by a scoring system and approximately 75% of malignant and 87% of benign ovarian tumors were correctly diagnosed preoperatively by digital subtraction angiography alone. Intravenous digital subtraction angiography can be applied as an easy, safe, and valuable diagnostic method for ovarian malignancies and can replace conventional pelvic angiography. PMID- 3051872 TI - Septic abortion caused by Salmonella enteritidis. AB - Septic abortion caused by salmonella infection is rare. Here we report a case of septic abortion due to Salmonella enteritidis. The route of infection was probably transplacental. PMID- 3051871 TI - Detection of abnormal cervical smears. A comparative study. AB - The inefficiency of the Ayres Spatula in detecting abnormality in cervical cytology has been demonstrated recently in a number of trials. In a randomized single-blind trial we have compared the Multispatula with the Ayres Spatula in a group of 158 women who previously had abnormal smears and had been referred to colposcopy clinics in the Lewisham and North Southwark Health Authority of London. The quality of the smears, as assessed by the presence or absence of endocervical cells, revealed that the Multispatula (87.3% pick-up) produced significantly better smears (p less than 0.005) compared with the Ayres Spatula (54.4% pick-up). In 149 smears collected with the Multispatula, atypia was confirmed. However, in only 129 smears taken with the Ayres Spatula were abnormal smears detected (p decreases less than 0.005). We concluded that the Multispatula produces a better quality smear which results in a decreased false negative rate in comparison with the Ayres Spatula. PMID- 3051874 TI - Prostaglandin vaginal suppositories in non pregnant women requiring cervical dilatation prior to hysteroscopy. AB - A randomized doubleblind investigation of prostaglandin vaginal suppositories prior to hysteroscopy was undertaken in 30 non-pregnant women. PGE2 in 14 of 15 treated patients we found a softening and a dilatation of the cervical canal, but with a relatively high frequency of side effects-nausea, vomiting and diarrhea. No serious bleeding or side effects were observed. PMID- 3051873 TI - Pudendal block in vaginal deliveries. Mepivacaine with and without epinephrine. AB - Pudendal block with 20 ml 1% mepivacaine with and without epinephrine was performed in 151 patients during the second stage of labor. No differences in efficacy of the block or in Apgar scores between the two groups were found. The maternal mepivacaine concentration was higher in the plain group than in the epinephrine group (p less than 0.01), but toxic levels were never reached. In the infants, no difference in mepivacaine concentration was found between the groups (p greater than 0.05, type II error 9%) and toxic levels were not reached. The time elapsed from the pudendal block until delivery was prolonged when epinephrine was added (p less than 0.02). We found no effect on blood pressure in either of the groups, with or without oxytocin and/or methergin. Twenty ml 1% mepivacaine (plain) is a safe choice for pudendal block without the possible disadvantages of adding epinephrine. PMID- 3051875 TI - Ovarian cysts in women with inflammatory bowel disease. AB - The frequency of ovarian cysts in patients with Crohn's disease (CD) or ulcerative colitis (UC) is believed to be higher than in the normal population, but this aspect has not been studied hitherto. The prevalence of ovarian cysts in the normal population is unknown. By ultrasonic scanning, we studied the frequency of ovarian cysts in 61 patients with CD, 64 with UC, and in 100 controls. The findings were positive in 3 out of 61 with CD, 5 of 64 with UC, and in 2 of 100 controls. There is a tendency to a higher frequency of ovarian cysts in patients with inflammatory bowel diseases than in the normal population, but no statistically significant difference. PMID- 3051876 TI - A randomized trial of three copper IUDs (MLCu250, MLCu375 and Nova-T). AB - A randomized prospective trial of three copper IUDs, Nova-T, MLCu375 and MLCu250, including 200 of each, is presented. Insertion was done at the hospital outpatient clinic on normally menstruating women and on women in puerperio. Follow-up was scheduled after 12, 24 and 36 months. Pregnancy rates were low for all 3 models. Pearl indices after 3 years were 0.5, 0.9 and 0.8 for Nova-T, MLCu375 and MLCu250 respectively (NS). Abnormal bleeding and/or pain was the most frequent termination cause. Minor differences in the termination rates because of abnormal bleeding and/or pain were found and are discussed. The continuation rates based on all medically relevant IUD removals were 74%, 73% and 81% after 3 years for Nova-T, MLCu375 and MLCu250 respectively. No important difference in clinical performance between the three copper IUDs could be demonstrated. PMID- 3051877 TI - Normal placental function and fetoplacental blood circulation in advanced abdominal pregnancy. AB - Normal levels of unconjugated estriol and human placental lactogen hormone in the maternal serum and a normal, reactive pattern of non-stress test cardiotocography were recorded in a case of advanced abdominal pregnancy with surviving newborn. The placenta was inserted at the base and side walls of fossa Douglas. Fetal, umbilical and retroplacental arterial Doppler ultrasound examinations revealed normal velocity waveforms. These findings indicated that the trophoblast invasion to the retroplacental arteries also occurs when the placenta has an extra-uterine insertion, and this can result in a normal placental blood supply and function. PMID- 3051878 TI - Comparison between lysine vasopressin and a long-acting analogue (N alpha triglycyl-lysine vasopressin) used as local hemostatic agents for conization. AB - Lysine vasopressin and a long-acting analogue N alpha-triglycyl-lysine vasopressin were compared in a prospective randomized double-blind study including 71 women undergoing cold knife conization of the uterine cervix. Hemodynamic and hemostatic variables were studied. N alpha-triglycyl-lysine vasopressin had the following advantages over lysine vasopressin: it gave significantly less skin pallor, becoming evident at a later stage during the operation. The diastolic blood pressure was significantly lower, as also was the incidence of postoperative hemorrhages. Factor VIII related antigen was lower. On the other hand reduction in heart rate (values before conization compared with values during conization) was more pronounced when N alpha-triglycyl-lysine vasopressin was used, but there was no difference in absolute values between the two groups during conization. PMID- 3051879 TI - Treatment of premenstrual mastalgia with tamoxifen. AB - Thirty-four women with normal menstrual cycles but suffering from severe premenstrual mastalgia were randomly treated with either tamoxifen (10 mg daily) (18 women) or placebo (16 women) from cycle day 5 to 24 for six consecutive cycles. At the end of the treatment, 89% of the tamoxifen-treated patients were free from the symptoms and the remainder experienced partial alleviation. In contrast, six patients treated with placebo showed only partial alleviation (38%) (p less than 0.001). Twelve months after the end of tamoxifen treatment, 53% of the medicated women were still free of symptoms as compared with none of the placebo-treated patients. These results suggest that tamoxifen is highly effective for the management of severe premenstrual mastalgia and should be useful for the treatment of this disorder. PMID- 3051881 TI - Intrapartum management of insulin-dependent diabetes mellitus (IDDM) gestants. A comparative study of constant intravenous insulin infusion and continuous subcutaneous insulin infusion pump (CSIIP). AB - Two groups of insulin-dependent diabetes mellitus (IDDM) parturients were managed according to different intrapartum protocols and compared with a control group of normal women in labor. Thirty-seven patients received conventional intensified insulin therapy during pregnancy and intravenous continuous insulin infusion during labor. Twenty-eight women were managed by continuous subcutaneous insulin infusion pump (CSIIP) throughout gestation and this therapeutic approach was continued during labor. When the two glucose control techniques were compared, CSIIP was superior to achieving and maintaining intrapartum optimal metabolic control, reducing significantly the incidence of acute fetal distress, thus lowering the cesarean section rate and neonatal hypoglycemia. Consequently we concur with the recommendation that firm prepartum as well as intrapartum diabetic control is mandatory for successful management of labor in IDDM patients. PMID- 3051880 TI - The effect of clomiphene citrate and tamoxifen on the cervical mucus. AB - The anti-estrogenic effect of clomiphene citrate (CC) and tamoxifen (TMX) on cervical mucus was evaluated in a prospective crossover study. Ten women underwent randomized alternate cyclical treatment with either 100 mg CC, 40 mg TMX daily, or with placebo. The effect of CC, TMX and placebo on serum estradiol (E2), cervical mucus secretion and on the development of ovarian follicles was evaluated. Compared with placebo, treatment with CC and TMX significantly increased the number of mature ovarian follicles on the day of assumed ovulation (p less than 0.05), elevated E2 secretion (p less than 0.05) and decreased cervical score (p less than 0.05). It can be concluded that anti-estrogenic agents reduced the secretion of cervical mucus. However, in most cases, the excessive E2 production due to multiple follicular growth overcomes the anti estrogenic effect. PMID- 3051882 TI - The clinical significance of asymptomatic mid-trimester low placentation diagnosed by ultrasound. AB - A prospective investigation of the problem of low placentation in mid-trimester, in uncomplicated pregnancies was performed. 773 pregnant women were included and 70 (9.1%) had a low-positioned placenta, defined as a placenta reaching or partially covering the cervical os, diagnosed by ultrasound between the 19th and 21st week of pregnancy. In no case did the placenta completely cover the cervical os. By repeat ultrasound examination in the 36th week of pregnancy, all low placentations had converted to normal position. Low placentation in mid-trimester was not associated with pregnancy complications or neonatal complications. A placenta reaching or partially covering the cervical os, in early pregnancy, seems to be normal, not influencing the subsequent course of pregnancy. PMID- 3051883 TI - Umbilical artery and uteroplacental blood flow velocity waveforms in normal pregnancy--a cross-sectional study. AB - Umbilical and arcuate artery blood flow velocity waveforms (FVW) were recorded in 125 normal singleton pregnancies from 20 to 42 weeks of gestation. The FVW were analysed for pulsatility index (PI), peak systolic velocity/minimum diastolic velocity ratio (S/D ratio), rising slope (RS) and descending slope (DS). Both in the umbilical and arcuate arteries, values for all variables declined with advancing gestation, indicating decreasing placental vascular resistance. The umbilical artery PI was unaffected by the fetal heart rate, but the arcuate artery PI was negatively correlated to the maternal heart rate (r = -0.40). The arcuate artery PI decreased by 0.00394 with each beat per minute increase in maternal heart rate. Normal limits (mean +/- 2 SD) were established for umbilical artery PI corrected for gestational age, and arcuate artery PI corrected for gestational age and maternal heart rate. PMID- 3051884 TI - Life-threatening hemorrhage due to uterine vascular abnormality. AB - A case is reported of a 25-year-old woman stricken with prolonged and life threatening menorrhagia from abnormal uterine vessels resembling hemangioma cavernosum. The condition was suspected at ultrasonic investigation. Hysterectomy was performed as an emergency operation. PMID- 3051885 TI - Seventeen week pregnancy in a rudimentary uterine horn revealed at routine ultrasonography. AB - Pregnancy in a rudimentary horn of a bicornuate uterus is a rare and often fatal event. A case is presented of such a pregnancy detected in the second trimester routine ultrasonographic examination. PMID- 3051886 TI - Observations on cochlear blood flow using the laser Doppler method in guinea pigs. AB - Cochlear blood flow of the guinea pig was measured using the laser Doppler method. Blood pressure and respiratory rhythm were also recorded simultaneously. The reliability of this method was substantiated by comparing it with the hydrogen clearance method using rabbit auricles. The blood flow of the auricle was altered artificially by clipping these vessels and then simultaneously measuring the blood flow by the two methods (Doppler and hydrogen clearance). The data obtained by these two methods were closely correlated. Norepinephrine induced both an elevation of blood pressure and an increase in cochlear blood flow in the guinea pig. Phentolamine, on the other hand, induced a fall in blood pressure and a slight decrease in cochlear blood flow. In addition, phentolamine completely blocked the effects of norepinephrine on blood pressure and cochlear blood flow. In this experiment, the responses of blood pressure and cochlear blood flow induced by norepinephrine and phentolamine were dose-related. PMID- 3051887 TI - Audiological findings after stereotactic radiosurgery in acoustic neurinomas. AB - Stereotactic radiosurgery was used in the treatment of 126 patients with acoustic neurinomas up to 30 mm in diameter from 1969 to 1984. Adequate follow-up data (mean follow-up period 4.7 years) were available for 111 (116 ears) of these 126 patients; of these 111 patients, 64 (65 ears) had a pure-tone threshold of less than 90 dB before the treatment and were followed up audiologically. Preserved hearing was found in 26% of the ears one year postoperatively. Shrinkage of the tumour was obtained in 44% and arrest of its growth in 42%. There was no mortality related to the radiosurgical treatment. Transitory facial weakness was noted in 15% of the patients (3% in 1983-84). Eighteen per cent of the patients had some, usually transitory, trigeminal dysfunction. The stapedius reflex threshold was improved in 13 ears (20%). In one patient the audiological tests became pathological in the contralateral ear during growth of a new tumour. Initially the stapedius reflex threshold was elevated, and 11 months later the BRA pattern also became abnormal. PMID- 3051888 TI - Lymphocyte subsets of maxillary mucosa in chronic inflammation. AB - Subsets of infiltrating lymphocytes within maxillary sinus mucosae of patients with chronic sinusitis were investigated by immunoperoxidase staining of frozen sections with the use of monoclonal antibodies. The most commonly observed infiltrating cell type was suppressor/cytotoxic T cells (CD8+) and smaller subpopulations of lymphocytes were helper/inducer T cells (CD4+) and B cells (CD20+). Variable numbers of HLA-DR+ cells were commonly observed in the lamina propria. The fibrous type of chronic sinusitis was found to have more suppressor/cytotoxic T cells (CD8+) and lower CD4/CD8 ratio than the other histopathological types. PMID- 3051889 TI - [Meta-traumatic laryngo-tracheal stenosis: clinical picture, therapeutic directions and critical evaluation from personal experience]. PMID- 3051890 TI - [Projective methods and dangerous behavior]. AB - The prediction of dangerousness by projective methods deals with some specific problems related to the bound between projective material and overt behaviour. PMID- 3051891 TI - [Psychological aspects associated with the diagnosis and treatment of cancer]. AB - Cancer and its treatments have numerous psychological consequences for patients. The consequences are related to the phase and the type of illness, and to the type and duration of the treatments. All these factors influence the apparition of adjustment difficulties or disorders. The present article review actual concepts and data related to the psychological problems secondary to cancer, its evolution and treatment. PMID- 3051892 TI - Assessing and managing hyperlipidemia. AB - Because more than one-half of adult Americans have total blood cholesterol levels that often contribute to atherosclerotic blockage of their coronary arteries, routine random screening of all adults and high-risk children for hypercholesterolemia is recommended. Reduced intake of saturated fat and cholesterol can lower total and low-density lipoprotein (LDL) cholesterol by 10 20 percent, while several medications lower total and LDL cholesterol by 15-40 percent. A highly effective cholesterol-lowering medication, lovastatin, has been recently marketed. The efficacy and long-term safety of ingesting large amounts of omega-3 fatty acids in fish oil supplements are unproven. Hypercholesterolemia is a family problem transmitted between generations by various combinations of genetic factors and learned behaviors. The family physician can be most effective by working with entire families to detect and treat hypercholesterolemia early in life to prevent serious consequences of prolonged cholesterol elevation. PMID- 3051893 TI - Physical standards for scuba divers. AB - Scuba diving has become a popular aquatic sport during the past 2 decades, and family physicians are frequently involved in examining scuba divers and in the decision making that allows them to pursue their training or careers in this sport. The purpose of this article is to review the physiology and gas laws that are involved in diving and to provide guidelines for assessing each diving candidate. The clinical manifestations of decompression sickness are discussed as well as the medical problems that could cause severe morbidity or mortality if diving is attempted. PMID- 3051894 TI - Ventricular arrhythmias, Part II: Special concerns in evaluation. AB - In the initial installment of this series, the prevalence, significance, and indications for treatment of ventricular arrhythmias were discussed. In this section, we address special concerns in the evaluation of patients with ventricular arrhythmias including the importance of extracardiac and exacerbating factors, the likely role of silent myocardial ischemia, benefits and drawbacks of monitoring methods, proarrhythmic effect, and torsades de pointes. Treatment considerations and discussion of pitfalls in management will follow in the final two installments of this series. PMID- 3051895 TI - Abdominal complications of infectious mononucleosis. AB - Infectious mononucleosis, a systemic illness caused by the Epstein-Barr virus, is seen frequently by primary care physicians. Mononucleosis affects several organ systems, and, within the abdomen, there can be splenic involvement, hepatitis, mesenteric lymphadenopathy, hyperplasia of gut-associated lymphoid tissue, pancreatitis, and transient malabsorption. Life-threatening abdominal complications require prompt recognition and intervention. Other abdominal complications, though worrisome, are usually short-lived and resolve without sequelae. PMID- 3051896 TI - Sarcoidosis: current concepts and case reports. AB - Sarcoidosis is a systemic granulomatous disease of unknown etiology associated with various immune alterations and biochemical changes. This article reports recent advances in the conceptualization of the immune dysfunction with emphasis on helper T-cell overactivity in the lungs. Because 90 percent of patients with sarcoidosis have intrathoracic disease, the mode of presentation, radiographic findings, clinical course, and treatment of pulmonary involvement are discussed. Case reports are used to demonstrate the typical course of the disease and generally favorable outcome of the vast majority of patients seen in the non referral setting. A rare case of neurosarcoidosis with neuroendocrine features is presented. PMID- 3051897 TI - Cell kinetics and multimodal prognostic evaluation in glial tumours investigated by serial stereotactic biopsy. AB - A retrospective evaluation of the prognostic value of different parameters available in patients affected by glial tumours and submitted to serial stereotactic biopsy is presented. The series investigated includes thirty-three untreated patients with proven brain gliomas submitted to stereotactic biopsy. All patients have been clinically and neuroradiologically monitored for three years. The factors investigated belong either to the preoperative data (clinical history and symptomatology, CT pattern and volume of the lesion) or to histological and biological data obtained after the stereotactic biopsy. The results suggest the need of a multimodal prognostic evaluation in glial tumours and particularly stressed is the accuracy of prognostic indications derived from cell kinetic studies. PMID- 3051900 TI - A study of the P300 component during minor hypoglycaemia. PMID- 3051898 TI - Meningeal melanocytoma. Report of a case and review of the literature. AB - A primary localized, partly encapsulated melanotic tumour broadly attached to the occipital dura and tentorium was removed from a 27 year-old woman. It recurred almost four years later in the posterior fossa and was resected again. Although grossly resembling a meningioma, the tumour lacked the histological, immunocytochemical and ultrastructural features of meningiomas and was characterized by the presence of numerous melanosomes and premelanosomes in the cytoplasm of tumour cells and macrophages and was, therefore, classified as "meningeal melanocytoma". The clinical and pathological features of this and 15 other cases in the literature are reviewed. The neoplasm, mainly occurring in the posterior cranial fossa and spinal canal, may cause neurological deficits through expanding, but non-invasive growth. Its biological behaviour is variable, and recurrence may occur after incomplete resection, but transition into malignant melanoma has not been observed. Total resection of this rare pigmented tumour arising from the pial melanocytes is recommended. PMID- 3051899 TI - New head fixation for the Riechert stereotaxic system. Technical note. AB - The combined use of stereotaxic and microsurgical techniques makes it possible to minimize damage to critical nervous tissue during operations in subcortical regions. The Riechert stereotaxic system has been further modified. The patient's head is fixed in the new head ring with standard Mayfield pins. The headring is connected to a standard Mayfield clamp at symmetrical bearings at 0, 90, 180, and 270 degrees, which holds the head stable in any desired position and allows unhindered access to the cranial vault and skull base. PMID- 3051901 TI - P300 and memory in adolescents with common migraine. PMID- 3051902 TI - Sex differences in cerebrovascular disease and its association with carotid obstruction. Retrospective evaluation of 1003 patients by Doppler examination. PMID- 3051903 TI - Chronic subdural hematoma associated with dural metastasis from mammary carcinoma. Case report and review of the literature. PMID- 3051904 TI - Primary spinal epidural lymphomas. PMID- 3051906 TI - [Renal angiomyolipoma. Diagnostic and therapeutic aspects]. PMID- 3051905 TI - [Cancer of the urinary bladder in Cartagena county]. PMID- 3051907 TI - [Urologic complications in 280 renal transplants]. PMID- 3051908 TI - [Nuclear medicine in the diagnosis of vascular and urologic complications of kidney transplantation]. PMID- 3051909 TI - [Comparative study of the usefulness of various perfusion indices in the diagnosis of acute rejection crisis in renal transplantation]. PMID- 3051910 TI - [Should systematic correction of vesicoureteral reflux be performed before renal transplantation?]. PMID- 3051911 TI - [Urothelial carcinoma of the upper urinary tract associated with hydronephrosis caused by stenosis of the pyeloureteral juncture, in a patient with a single functioning kidney. Apropos of a case. Review of the literature]. PMID- 3051912 TI - [Substitution of the ureter with ileum in renal transplantation]. PMID- 3051913 TI - Smoking as an issue in alcohol and drug abuse treatment. AB - Little attention has been given to the role of tobacco dependence within alcohol and drug abuse treatment. Yet, smoking behavior appears to be interrelated with the use of alcohol and other drugs. This interrelationship is explored, and the role of smoking cessation within alcohol and drug abuse treatment is considered. Areas for future research on this topic are identified. Addictive disorders are generally thought to include alcohol abuse, drug abuse, smoking, overeating, and, sometimes, gambling and caffeine dependence. While some attention has been paid to the common etiological roots of various addictive disorders, relatively little systematic attention has been paid to commonalities in their treatment and especially to the treatment of multiple disorders in the same individuals. The one significant exception is alcohol abuse and drug abuse. Of the other addictive disorders, tobacco dependence has been most closely interrelated with alcohol and drug abuse. Yet, little attention has been given to tobacco dependence within alcohol and drug abuse treatment. This paper will focus on smoking in relationship with alcohol and drug abuse, and will consider the possible role of smoking cessation treatment within the context of alcohol and drug abuse treatment. First, background regarding the interrelationship of alcohol and drug abuse is explored. Then, the relationship of smoking with other substance use is considered, followed by a review of special concerns related to smoking among alcohol and drug abuse clients. Next, the current status of smoking cessation within alcohol and drug abuse treatment is addressed. Finally, implications are considered. PMID- 3051914 TI - Antibody combining sites: how much of the antibody repertoire are we seeing? How does it influence our understanding of the structural and genetic basis of antibody complimentarity? PMID- 3051915 TI - Mannose-binding proteins of animal origin. PMID- 3051916 TI - Differential binding properties of Ga1NAc and/or Ga1 specific lectins. AB - Grouping of lectin binding properties, based on determinant structure rather than monosaccharide inhibition pattern, should facilitate the selection of lectins as structural probes for glycans as well as for the interpretation of the distribution and the properties of the carbohydrate chains on the cell surface. Based on the binding specificities studied with glycan by precipitin-inhibition, competitive-binding and hemagglutinin-inhibition assays, twenty Ga1 and/or Ga1NAc specific lectins have been divided into six classes according to their specificity for the disaccharide as all or part of the determinants and Ga1NAc alpha 1----Ser(Thr) of the peptide chain. The differential affinities of these lectins were characterized by quantitative precipitin assay. Abbreviation of the following six lectin determinants can also be used to classify these lectins. (1) F determinant (GalNAc alpha 1----3GalNAc, Forssman specific disaccharide). (2) A (Af) determinant (GalNAc alpha 1----3Gal, Human blood group A specific disaccharide; Af, fucosylated A, (GalNAc alpha 1----3 [LFuc alpha 1----2]Gal). (3) Tn determinant (GalNAc alpha 1----0 to Ser (Thr) of the protein core, Tn antigen). (4) T determinant (T antigen, Gal beta 1----3GalNAc alpha 1----0 to Ser (Thr) of the protein core, the mucin type sugar sequence on the human erythrocyte membrane or Gal beta 1----3GalNAc beta 1---- at the nonreducing end of ganglioside). (5) I and II determinants (human blood group type I and II carbohydrate sequences). Most of the lectins reactive to Gal beta 1----4GlcNAc (II) are also reactive to Gal beta 1----3GlcNAc (I). Lectin I (II) determinants (i.e. Gal beta 1----3 (4) GlcNAc residues) can be found at the nonreducing end of the carbohydrate chains derived from either N-glycosidic or O-glycosidic linkages. (6) B determinant (Gal alpha 1----3Gal, Human blood group B specific disaccharide). Their carbohydrate specificities are classified as following: (Table see text). The differential binding properties of lectins can be defined from comparisons of their carbohydrate specificities listed above. PMID- 3051917 TI - Antigenic properties of human erythrocyte glycophorins. PMID- 3051918 TI - Structural concepts of the human blood group A, B, H, Le(a), Le(b), I and i active glycoproteins purified from human ovarian cyst fluid. AB - Regardless of the A, B, H, Le(a), Le(b), I and i activity, purified water-soluble blood group glycoproteins from human ovarian cyst fluid have a similar overall structure. They are polydisperse macromolecules (Mr 2.0 x 10(5) to several million) of similar composition (75 to 85% carbohydrate, 15 to 20% protein) and consist of multiple heterosaccharide side chains attached by an O-glycosidic linkage at their internal reducing ends to serine or threonine of the polypeptide backbone. About 90% of these carbohydrate side chains range in size from one to less than twenty-four sugar residues (twelve sugars in the internal structure and twelve key sugars specific as blood group determinants). Three-fourths of these side chains contain fewer than twelve sugars. A generalized blood group active carbohydrate chain is shown above. Three disaccharide units-Type I chain (Gal beta 1----3GlcNAc beta 1----3), Type II chain (Gal beta 1----4GlcNAc beta 1----6) and T determinant [Gal beta 1----3GalNAc alpha 1----Ser(Thr)]-are used to elucidate the internal structure of the carbohydrate chains. The complete internal structure is considered to have a core structure with four branches, to which the blood group key sugars are attached at the appropriate locations. The core structure is a tetrasaccharide, composed of one unit of Type I chain at the nonreducing end and the T determinant at the other end, linked to Ser or Thr of the protein moiety. Branch I is Type I chain and Branch II is Type II chain. They are linked to Gal at the nonreducing end of the core structure. Branch III is usually a Type II chain, but may sometimes be a Type I chain, linked to the GalNAc of the reducing end. The length of Branch III can be increased by adding one or more monosaccharides of Type I chain sequence such as Gal beta 1--- 3GlcNAc beta 1----3Gal beta 1----4GlcNAc beta 1----6GalNAc alpha 1----Ser(Thr), a combination of Type I and Type II chains. A new Branch IV is made up of Type II chain, which in turn is linked to the Gal end of the T determinant. The Type II chains react with the antibody to the type XIV pneumococcal capsular polysaccharide and with the anti-I(Ma) cold agglutinin.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3051919 TI - Biochemistry and lectin binding properties of mammalian salivary mucous glycoproteins. AB - The molecules responsible for the highly viscous properties of mucus are secretory glycoproteins referred to as mucins. Salivary mucins are characterized by a high sugar to protein ratio and are of a broad range of molecular weight from 7 x 10(4) to millions. With a few exceptions, they contain up to 30% of hexosamine (galactosamine and glucosamine), 8-33% of sialic acid, trace to 15% of galactose or fucose and little or no mannose. The size of carbohydrate side chains of these glycoproteins ranges from one to about fifteen units of sugar. These carbohydrate side chains are usually O-glycosidically linked through N acetylgalactosamine to a peptidyl serine or threonine. In some instances, ester sulfate groups, mainly on N-acetylglucosamine, are also a structural feature. In many of these glycoproteins, the saccharide sequence is the same as that which determines the specificity of blood groups. Carbohydrate sequence analysis shows that salivary mucins exhibit considerable polydispersity, great diversity and remarkable structural flexibility not only among animal species but also within the same mucin molecule. Based on their lectin-binding ability, they can be used for purification of lectins, and lectins coupled to resin may be useful for the isolation of mucin-type glycoproteins. The epithelial mucous secretions modulate oral microbial flora; many secretory components serve as lectin-receptors for the attachment of microbes. The judicious use of lectins with widely differing binding characteristics has already been valuable in the in situ localization of salivary glycoproteins, in elucidating structural details, recording sugar density within a given tissue section, and defining host-parasite interactions. It is hoped that their use, together with monoclonal antibody (158) and tissue culture techniques (159, 160) will further clarify the roles of individual secretory mucous glycoproteins in health and disease. PMID- 3051920 TI - New trends in ganglioside chemistry. AB - New methods have been developed for the preparation of highly purified gangliosides, homogeneous in the saccharide, long chain base, and fatty acid moieties and gangliosides carrying different kinds of labelled probes. Gangliosides, homogeneous in the oligosaccharide portion, were prepared by preparative normal phase HPLC on a Lichrosorb-NH-2 column, using a gradient of acetonitrile-phosphate buffer, pH 5.6, as solvent system. Each class of ganglioside (from monosialo- to tetrasialogangliosides) was then submitted to reversed phase HPLC on a preparative RP-8 column, using acetonitrile-5 mM phosphate buffer, pH 7, as solvent system, to obtain gangliosides homogeneous in the long chain base moiety. Gangliosides containing C18 and C20 sphinganine were prepared by catalytic hydrogenation of the corresponding unsaturated gangliosides. GM1 with homogeneous acyl chain was prepared by alkaline hydrolysis in the presence of tetramethylammonium hydroxide (which forms a GM1 deacetylated at the level of sialic acid, and a GM1 deacetylated at the level of sialic acid and deacylated at the level ceramide), followed by re-N-acylation, carried out in the presence of dimethylaminopropyl, ethylcarbodiimide and natural fatty acids, or of mixed anhydride of ethylchloroformate and 14C-stearic acid, and re-N acetylation performed with acetic anhydride or labelled acetic anhydride. The GM1 derivative, de-acetylated at the level of sialic acid, also produced by alkaline treatment of GM1, was submitted to re-N-acetylation with 14C-acetic anhydride to produce specifically 14C-labelled GM1. Re-N-acylation was carried out a) in the presence of dimethylaminopropyl, ethylcarbodiimide and natural fatty acids, b) with mixed anhydride of ethylchloroformate and 14C-stearic acid. After re-N acylations, re-N-acetylation was performed with acetic anhydride or labelled acetic anhydride. Gangliosides tritium labelled in the oligosaccharide moiety were prepared by the galactose oxidase/3H NaBH4 method, and gangliosides tritium labelled at carbon-3 of unsaturated long chain bases by the dicyano dichlorobenzoquinone (DDQ)/3H NaBH4 method. PMID- 3051921 TI - Chemical and immunochemical studies on lipopolysaccharides of Coxiella burnetii phase I and phase II. AB - Lipopolysaccharides of Coxiella burnetii phase I and II were comparatively investigated by chemical and immunochemical methods. LPS of phase I (LPS I) and phase II cells (LPS II) show no serological cross reaction, indicating that the serological determinants of LPS II are masked in LPS I. Chemical analysis of LPS I and II show that phase I and II cells can be considered as S and R forms of Coxiella burnetii. The structure of LPS II has recently been elucidated and shows a dimannosylated core of an alpha(1,3)-linked heptose-disaccharide which is attached to a "KDO-like" substance. In enterobacterial core-types, alpha(1,3) linked heptose-disaccharide is also part of the inner core structure, although the heptose occurring in enterobacterial R cores is the L-glycero-D-manno heptose. In Coxiella burnetii we have only the rare D-glycero-D-manno-heptose which is the biosynthetic precursor of the former and is in many enteric LPS, present only in addition to L-glycero-D-mannoheptose. In these R-cores, it is occupying mostly terminal positions (Radziejewska-Lebrecht et al., 1981) and is absent from the main chain. The complete structure of LPS I is not yet available, but some important points could recently be clarified. The immunodominant sugars in LPS I are C-3-branched sugars, 6-deoxy-3-C-methyl-L-gulose (L-virenose) and 3 C-(hydroxymethyl)-L-lyxose (dihydro-hydroxy-L-streptose). These two sugars have not been found so far in other lipopolysaccharides and the latter one not previously in any other natural product. Their identification is based on GLC-MS comparison with authentic and synthetic compounds. Both branched sugars (and in addition part of the mannose) are the terminal sugars in LPS I. Sites of attachment of phase I-specific sugars to the LPS II-core are: the 3-position of a branched heptose and, presumably, the 4-position of a terminal D-mannose. The extreme acid-lability of the linkages of both branched sugars was investigated in detail and is caused by the nature of the branched sugars (deoxyhexose with bulky axial substituents; pentofuranose with axial OH-groups). No information is so far available on the (penultimate) sugars to which the branched sugars are linked, but methylation analyses with LPS I, and with the recently described I/CR mutant, which is selectively lacking the virenopyranose, are presently performed. PMID- 3051922 TI - Tumor-associated blood group antigen expressions and immunoglobulins associated with tumors. AB - As outlined in Figures 1 and 2, the biosynthetic pathways for the expression of the A, B and H, and the Lewis determinant carbohydrate sequence structures, as well as sialylated structures, involves both type 1 and type 2 precursor chains (which may be present as glycolipids and N- or O-linked glycoproteins), and many glycosyltransferases. For tumor cells, there appears to be increased expressions of fucosyl- and sialyltransferases yielding such structures as the Le(x), sialyl Le(a), and many other similar determinants, which are not found on the normal cell progenitor of the tumor. The types of structures expressed on tumor cells is dependent on the particular fucosyl-, sialyl- and other glycosyltransferase genes activated in the transformation and tumor progression events, the availability of the substrates for the glycosyltransferases (both the precursor sequences and the nucleotide-sugar substrates) which is partly dependent on metabolites available to the tumor mass, and on the genotype of the individual regarding particular glycosyltransferases. Both the loss of A, B and/or H blood group antigen expressions of tumor cells and the relative expressions of the Lewis and sialylated-oligosaccharide determinants may be a consequence of the competing biosynthetic pathways and the glycosyltransferases for common substrate sequences, as well as due to the loss of particular glycosyltransferases concomitant with transformation. All of these factors probably account for the variable expressions of the complex of carbohydrate sequence determinants when comparing tumor sections of different individuals as well as the heterogeneity of expression of particular determinants within a single tumor tissue section. As described above, the A, B and/or H determinants, and the precursor sequences, are also expressed to differing extents on epithelial cells depending on the tissue type and cellular location in the tissue. Thus, the differentiation state of the particular epithelial cell also determines the quantity and types of carbohydrate sequences expressed. However, because of the complex nature of the competing biosynthetic pathways for the carbohydrate sequences of glycolipids and glycoproteins, and the relative activations of fucosyl- and sialyltransferases of tumor cells, it would seem that simple deductions as to the state of differentiation of particular tumors with A, B, H and precursor sequence expressions is not warranted.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3051923 TI - The chemical basis for expression of the sialyl-Le(a) antigen. AB - The SLe(a) antigen, originally defined by monoclonal antibody 19-9, is a complex carbohydrate epitope that differs from the normal human blood group Lea antigen only by the presence of an additional sialic acid residue. SLe(a)-active oligosaccharides occur in both gangliosides and mucin-like glycoproteins in developing embryonic gut, as well as in many normal adult glandular tissues and secretions, but the antigen is virtually absent from normal adult gastrointestinal lumenal epithelial cells. Following malignant transformation of adult gastrointestinal lining epithelium and many other endodermally-derived glandular epithelia, SLe(a)-active mucins released from the ensuing tumor appear in blood plasma. The level of circulating SLe(a) antigen is currently being investigated as a means of following tumor recurrence, progression, and therapy. Recent studies on the biosynthesis of SLe(a) explain the observations that, 1) the antigen does not occur in individuals of Le(a-b-) blood group, and 2) individuals that belong to the Le(a+b-) blood group express SLe(a) more strongly than Le(a-b+) individuals. Further, the biosynthetic studies predict a new tumor antigen, NeuAc alpha 2-3Gal beta 1-3GlcNAc beta 1.... (the immediate precursor to SLe(a)) that should be expressed in Le(a-b-) individuals in nearly the same tissue distribution as found for the SLe(a) antigen in Le(a+b-) and Le(a-b+) individuals. Based upon studies of SLe(a) expression in normal saliva and the pathway for biosynthesis of SLe(a), it seems likely that future clinical studies could be profitably directed towards improving the predictive value of the plasma SLe(a) level by adjusting the quantitative results according to the Lewis blood group and ABH secretor phenotype of the individual patient. PMID- 3051924 TI - Glycoconjugates and tumor metastasis. PMID- 3051925 TI - Structural elucidation of complex carbohydrates. AB - Many current problems in immunology depend on a detailed understanding of oligosaccharide and glycoconjugate structures. In contrast to other biopolymers, oligosaccharides do not have well established and sensitive analytical procedures for the determination of their structures. Additionally, oligosaccharides because of branching, linkage, and structural isomerism, pose specific complications unique to this biopolymer. We have discussed MS data utilizing DCI and FAB ionization and presented new information that indicates FAB-MS can provide a wealth of structural information including sequence, branching and molecular weight. In the final sections FAB ionization and collision induced dissociation, have been discussed, two adjunct techniques that amplify the analytical value of MS instrumentation in the pursuit of oligosaccharide structure. PMID- 3051926 TI - Blood group active haptens in urine and faeces. PMID- 3051928 TI - A guide for carbohydrate specificities of lectins. PMID- 3051927 TI - Analysis of complex carbohydrate primary and secondary structure via two dimensional proton nuclear magnetic resonance spectroscopy. PMID- 3051929 TI - The separation of immunocyte subpopulations by use of various lectins. AB - The use of lectins for the enrichment of various lymphocyte subpopulations was investigated. Bauhinia purpurea lectin (BPA) was found to be effective for the enrichment of B cells, and the B cells thus obtained were further fractionated with lentil lectin into subsets showing high and low responses to dextran sulfate. The ability of various lectins to selectively induce suppressor T cell activity or helper T cell activity was also examined. The suppressor T cells thus induced were enriched peanut lectin and, conversely, the helper T cells were enriched with Limulus polyhemus lectin. This method was applied to analysis of age-dependent changes in the levels of suppressor and helper T cells in autoimmune-prone mice. Cytotoxic T cells induced in an allogeneic mixed lymphocyte culture were enriched with Dolichos biflorus lectin (DBA). These cytotoxic T cells showed a specific killing effect in vitro. However, when spleen cells of tumor-bearing mice were fractionated by use of DBA, the DBA- cells mediated the regression of the tumors in vivo. BPA was also found to be effective for the enrichment of the interleukin-2-producing T cell subset and macrophage precursor cells. Using this technique, bone marrow cells of autoimmune-prone MRL/1 mice were found to be rich in macrophage precursor cells. PMID- 3051930 TI - Effect of des arginine9-bradykinin and other bradykinin fragments on the synthesis of prostacyclin and the binding of bradykinin by vascular cells in culture. AB - Bradykinin (BK) fragments, des arg1-BK, des arg1,pro2-BK, des phe8,arg9-BK and des pro7,phe8,arg9-BK were synthesized and along with des arginine9-BK (daBK), tested for their ability to induce prostacyclin synthesis in homogeneous cultures of cells from the calf pulmonary artery. Of the fragments daBK was the only peptide, in addition to bradykinin (BK), to activate the synthesis of prostacyclin (PGI2) and platelet activating factor (PAF) in endothelial cells and PGI2 in fibroblasts and smooth muscle cells. Half-maximal activation of PGI2 synthesis differed with the cell type. The other fragments tested did not directly affect PGI2 synthesis. These fragments also did not inhibit daBK or BK activation of PG synthesis. BK bound to endothelial cells with a dissociation constant (Kd) of 2.1 nM and a Bmax of 47.9 fmoles/10(6) cells. The Kd for the binding of BK to smooth muscle cells and fibroblasts was somewhat higher, 4.9 nM and 7.9 nM, respectively. None of the fragments tested, including daBK, altered the binding of BK. Des arg9[leu8]-BK, reported to be a competitive antagonist of the bradykinin B1 receptor, inhibited daBK induced PG of PAF synthesis in endothelial cells but had little effect of BK binding or BK induced PG synthesis. Finally, the BK antagonist [thi5,8, d-phe7]-BK blocked both BK binding and the ability of either BK or daBK to induce PG synthesis, thus substantiating that the binding of these kinins is a step in the activation of PG synthesis. PMID- 3051932 TI - Drugs affecting pulmonary responses to platelet activating factor as novel anti asthma drugs. PMID- 3051931 TI - Neutrophil migration induced by inflammatory stimuli is reduced by macrophage depletion. AB - Previous experiments of our group have shown that neutrophil migration induced by inflammatory stimuli is reduced by agents which block the release from macrophages of a specific factor for neutrophil migration (MNCF, [1, 2]). The present paper evaluated the influence of macrophage depletion induced by lavage of the peritoneal cavity on neutrophil migration. In both normal and thioglycollate-stimulated peritoneal cavities, lavage with saline reduced the resident macrophage population by about 80% and significantly blocked neutrophil migration induced by inflammatory stimuli such as carrageenin, zymosan and E. coli endotoxin. Peritoneal lavage, however, did not affect neutrophil migration induced by MNCF. Thus, these results support the suggestion that macrophages participate in the control of neutrophil migration induced by acute inflammatory stimuli. PMID- 3051933 TI - Airway microvascular permeability in asthma: a target for drug action? AB - Evidence is presented that inflammatory mediators likely to be involved in asthma can increase the permeability of airway microvessels to macromolecules (i.e. cause leakage). The microvessels involved are in the tracheobronchial circulation, which supplies the trachea and bronchi. The acute response to these mediators has been shown to involve interendothelial gap formation in postcapillary venules. In animals, leakage can be demonstrated histologically using macromolecular tracers such as colloidal carbon. Alternatively, it can be detected by measuring the amount of radiolabelled, or fluorescein-labelled, plasma protein, or similar macromolecules, in airway tissue and/or airway lumen. There is clinical evidence to support a leakage response in asthmatic lung. Data are available suggesting that anti-asthma drugs can decrease airway microvascular leakage in animals and this can also be deduced from some of the clinical studies in asthmatics. The importance of airway microvascular leakage, and hence of plasma exudate, in the pathophysiology of asthma is stressed and possible ways of attenuating it with drugs are summarised. PMID- 3051934 TI - Developments in anti-asthma glucocorticoids. AB - Studies carried out by Swedish and English workers in the mid 1950s appear to be the first successful uses of topical glucocorticoids in asthma. As shown with beclomethasone dipropionate during the 1970s, and expanded with budesonide during the 1980s, inhaled glucocorticoids have a very broad clinical efficacy and are safe to use. With increasing lung selectivity glucocorticoid drugs may well become a primary treatment for asthma. Even prolonged treatment with large doses of budesonide may not affect the contractile and relaxant characteristics of airway smooth muscle suggesting that this tissue is not a direct target for glucocorticoid actions. In IgE-sensitized guinea-pigs, as in atopic asthmatics, inhalational budesonide and other glucocorticoids inhibit both immediate and late phase pulmonary reactions occurring after allergen provocation. The anti anaphylactic lung effects of glucocorticoids are not necessarily associated with inhibition of histamine and leukotriene release and, although PAF release was reduced by budesonide, the importance of this effect is as yet unknown. Airway eosinophilia is seen at the late phase reaction but may not consistently be reduced by glucocorticoids. These drugs reduce plasma leakage that is associated with both atopic and non-atopic asthma. Hence, a stabilising effect on airway endothelial-epithelial barriers may be one of the significant actions of glucocorticoids in the inflamed airways of patients with asthma or chronic obstructive pulmonary disease. PMID- 3051935 TI - Action of mediators on airway smooth muscle: functional antagonism as a mechanism for bronchodilator drugs. AB - The beta-adrenoceptor agonists have become the cornerstone of bronchodilator therapy. These agents are "functional" or "physiologic" antagonists that actively relax airway smooth muscle through a cyclic-AMP (cAMP)-mediated decrease in myoplasmic Ca2+ content. Hence, unlike specific receptor antagonists, the sympathomimetics should reverse bronchoconstriction regardless of the mediator(s) involved. Indeed, one of the primary beneficial attributes of beta-adrenoceptor agonists is their inhibitory activity against a wide range of bronchoconstrictors. As successful as the sympathomimetic bronchodilators have been, they are not without liabilities. These liabilities include: 1) cardiovascular and skeletal muscle side effects, 2) an inherent subsensitivity of the patient population to beta-adrenoceptor agonists, 3) the development of tolerance, and 4) loss of efficacy during severe asthmatic episodes. These limitations are not specific for individual agents but are shared by all beta adrenoceptor agonists. A significant improvement in the pharmacotherapy of asthma would be obtained by identifying novel bronchodilators devoid of one or more of the aforementioned liabilities. The development of isozyme-selective phosphodiesterase (PDE) inhibitors is one promising approach toward this goal. Interest in PDE inhibition as a therapeutic target has been renewed by the realization that PDEs exist in multiple isoforms and that the distribution of these isoforms varies significantly among tissues. This information, coupled with the recent synthesis of PDE inhibitors selective for several of the isozymes, raises the possibility of breeding organ-selectivity into this class of compounds. Results from preliminary experiments with isozyme-selective PDE inhibitors have helped to identify appropriate drug targets in airway smooth muscle. These early studies suggest that the synthesis of novel isozyme-selective PDE inhibitors not only may provide tools with which to understand the biologic function of various PDE isozymes, but may also lead to the development of improved therapeutic agents. PMID- 3051936 TI - Calcium ion mechanisms in airway smooth muscle: potential targets for novel symptomatic drugs for asthma. AB - Calcium ions (Ca2+) are fundamental to the processes responsible for the initiation and maintenance of contraction of airway smooth muscle cells. Recent developments in our understanding of signal transduction mechanisms relating to intracellular Ca2+ release have extended our knowledge of excitation-contraction coupling mechanisms in airway smooth muscle. Furthermore, these developments open up potential targets for the development of new drugs, with novel mechanisms of action, for the symptomatic treatment of asthma. This article reviews these recent advances and focusses upon those calcium ion mechanisms that are possible targets for drug action. PMID- 3051937 TI - A review of 500 percutaneous nephrolithotomies. PMID- 3051938 TI - [A clinical evaluation of the protective effect of fosfomycin (FOM) against the cis-diamminedichloroplatinum (CDDP)-induced nephrotoxicity]. AB - The protective effect of fosfomycin (FOM) against the cis diamminedichloroplatinum (CDDP)-induced nephrotoxicity was evaluated clinically. Thirty-six cases suffering from urogenital cancers were subjected to CDDP therapy in which 50 mg of CDDP was administered on Day 0, with one week regarded as one course of treatment. The daily excretions of urinary N-acetyl-beta-D glucosaminidase (NAG) and beta 2 microglobulin (beta 2 MG) levels were measured on Day 1, 3 and 5. Creatinine clearance (Ccr) was also determined on Day 1. CDDP was administered once a week for 6 weeks. Group A consisted of 18 patients who were treated with CDDP alone every alternate odd week. In the other even weeks, they received both FOM 4.0 g and CDDP on Day 0 and FOM 4.0 g on Day 1 and 2. The maximum values together with the total amounts of urinary NAG and urinary beta 2 MG were measured. Group B, also consisting of 18 cases, was divided into two sub groups, one which was always administered with CDDP alone and another which received both FOM and CDDP. The maximum values of each course in urinary NAG and beta 2 MG were measured and Ccr level was also obtained. The changing pattern in the maximum values of two parameters and Ccr level between two subgroups was compared. Group A: The maximum values of urinary NAG excretions obtained in 4th week, using the combined therapy of CDDP and FOM was significantly lower than that obtained in the 3rd week.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3051939 TI - [Single day treatment in acute cystitis]. AB - The effect of a single day treatment with 600 mg norfloxacin 600 mg ofloxacin or 1,920 mg trimethoprim-sulfamethoxazol was determined on 114 patients with acute cystitis. The overall clinical efficacy was excellent in 101 patients (89%), moderate in 9 patients (8%) and poor in 4 patients (3%). Recurrence was observed in 8 cases (8%) within 6 weeks after the treatment. The effectiveness rate and the recurrence rate were inferior in those caused by S. epidermidis compared with those caused by E. coli. PMID- 3051940 TI - [Adrenal myelolipoma: two case reports]. AB - We report 2 cases of adrenal myelolipoma which were suspected preoperatively and confirmed by surgical resection. As with most cases previously reported, the lesions were found in obese, middle-aged persons. Laboratory tests of adrenal function revealed values within the normal level. Excretory urography showed radiolucent masses in the suprarenal area displacing the kidney inferiorly. Computerized tomography revealed well-defined masses consisting of fat density areas and higher density areas. Ultrasonography demonstrated hyperechoic, heterogeneous tumors. The tumors were shown to be hypovascular and adrenal in origin on the angiogram. An operation was performed because of complaint of flank pain in the first case and hypertension associated with elevated plasma renin activity in the second case. The pathological study disclosed typical adrenal myelolipoma with a mixture of hematopoietic and adipose tissue. Adrenal myelolipoma is clinically unusual and only 41 cases with premortem diagnosis have been reported in English literature. We herein report the 14th and 15th cases in Japan. As computerized tomography and ultrasonography become more widely used, we believe that the number of cases difficult to differentiate from a malignancy will increase. PMID- 3051941 TI - [A case of polycystic kidney with bilateral nephrectomy]. AB - A 56-year-old woman in chronic hemodialysis had been suffering from uncontrollable fever for the past 7 months. Her original disease was diagnosed as familial polycystic kidney and three of her five brothers were found to have the same disease. Her chromosome was 46,XX,21P+ and laboratory examination revealed severe anemia, malnutrition, liver dysfunction, pyuria and candidiasis of urine. Abdominal echogram and CT scan revealed polycystic kidneys and multiple liver cysts. She was admitted to our hospital and was diagnosed as having pyelonephritis of the right kidney. As her condition was not improved by conservative therapy right nephrectomy was performed. One month later, spiking fever and left tenderness reappeared. Those symptoms could not be controlled by conservative therapy and left nephrectomy was performed again. Pathological examination on nephrectomized kidneys showed interstitial nephritis, hyaline degeneration and proliferative change of glomeruli, microabscess, colloid of tubules and calcification of parts of Henle's loops. Nephrectomy has been performed in 1.6 to 10.0% of polycystic kidneys due to references since 1952. Eight of the 22 polycystic kidneys (36.3%) seen at our hospital during the past 10 years have been removed. PMID- 3051943 TI - [Ureteritis cystica: a case report]. AB - This is a case report on a patient with left renal stone and ureteritis cystica. The patient was a 57-year-old male, and he was admitted for a thorough examination of his renal stone. Many sharply-defined radiolucent filling defects were detected in the upper and middle ureter by retrograde pyelography, and he was diagnosed with ureteritis cystica. Since the patient had severe renal function damage, left nephro-ureterectomy was performed. This is the 29th case of pyeloureteritis cystica reported in Japan, and here in its clinical aspects are discussed. PMID- 3051942 TI - [A case of double cancer: renal pelvic transitional cell carcinoma and lung squamous cell carcinoma]. AB - A 69-year-old woman was admitted with the chief complaint of gross hematuria and left flank pain ten years after curative right pneumonectomy. Retrograde pyelography showed a filling defect of inferior calyx. Computerized tomography revealed a solid tumor with a low density area arising from the left kidney. The tumor was demonstrated hypovascular by angiography. Left radical nephrectomy by a transabdominal approach was performed. Histological diagnosis was primary transitional cell carcinoma of the left renal pelvis largely replacing the renal parenchyma. Twenty six days after the operation she was discharged. Our case was of double cancer consistent with Warren and Gates criteria and was classified into the nonsimultaneous case according to Moertels criteria. Double cancer of the lung and renal pelvis is very rare and our case seems to be the 7th in the Japanese clinical literature. PMID- 3051944 TI - [Epidermoid cyst of the scrotum: report of a case]. AB - A 61-year-old male visited our hospital with the chief complaint of a mass in the right scrotum on October 4, 1985. Under the diagnosis of intrascrotal tumor, the mass was resected surgically. It was a subcutaneous tumor, and had no relation to any intrascrotal organ, such as testis, epididymis or spermatic cord. The mass was 7 x 6 x 5 cm. Pathological diagnosis was epidermoid cyst of the scrotum. Epidermoid cyst of the scrotum is a rare disease and only 7 cases have been reported in Japan. PMID- 3051945 TI - [Citrate (CG-120) therapy for urolithiasis. 1. Clinical effects]. AB - Urinary citrate is an important determinant for crystallization of calcium salts, and recently oral administration of citrate has been suggested to be clinically useful in the management of renal stone disease. The effect of CG-120, a citrate compound (potassium citrate, sodium citrate and citric acid) produced by Dr. Madaus (Germany), on upper urinary tract stones was investigated in 398 patients in this study group. The patients were treated with 3 or 4 g CG-120 daily. Two hundred thirty-one of them were treated accurately according to the protocol of the study for more than 32 weeks. The cumulative percentage of positive clinical effect for the stone (disappearance, passage or decrease in size) was 30.3% (70/231). There were no differences in the clinical effect between the group of 3 g/day and the group of 4 g/day. CG-120 seemed to be more effective in the cases of ureteral stone, young patients and females, but was less effective in the recurrent stone formers. Although there were 45 episodes of side effects in 38 patients in this study, no serious side effects attributed to CG-120 were experienced. CG-120 was proved as a useful drug in the treatment of upper urinary tract calculi as well as its prevention. PMID- 3051946 TI - [Citrate (CG-120) therapy in urolithiasis. 2. Effect on urinary and serum biochemistry ]. AB - The long-term effects of citrate therapy (CG-120, 3 g/day or 4 g/day) were examined in 398 patients with upper urinary tract calculi. We studied the influence of citrate therapy on urinary and blood biochemistry in 353 of them. CG 120 caused a sustained increase in urinary citrate, urinary pH and potassium, but no substantial or significant changes in other urinary parameters (uric acid, phosphate, oxalate, sodium, chloride and urine volume). Although urinary calcium decreased significantly up to the 24th week, it did not change significantly there after and it tended to increase at the 54th week. Urinary creatinine excretion decreased after 34 weeks of administration, but this phenomenon could not be explained, because the level of blood urea nitrogen and serum creatinine was not elevated in any case before administration. There were no changes in the serum calcium, magnesium, phosphate, uric acid, sodium, potassium or chloride level. PMID- 3051947 TI - [Pharmacokinetic study on oxalate in rats]. AB - The pharmacokinetics of oxalate were studied in normal and nephrectomized rats, using radioisotope-labelled oxalate to resolve the mechanism of calcium oxalate stone formation. Plasma disappearance of 14C-oxalate was analyzed with a 2 compartment open model, and each compartment volume, the first-order rate constant for elimination, and the first-order rate constant for transfer between the central compartment and peripheral compartment were obtained. These values were compared with those for inulin. In normal rats, the total distribution volume was 57% of body weight for oxalate and 34% for inulin. The elimination rate constant from the central compartment for oxalate was lower than that for inulin, but oxalate had a much larger central compartment volume than inulin. Thus, the total clearance of oxalate was greater than that of inulin. In nephrectomized rats, total clearance of oxalate decreased to 1/6 of that in normal rats, while total clearance of inulin decreased to 1/27 of that in normal rats. These results suggest that oxalate is more diffusible than inulin, and that oxalate is excreted mainly from the kidney. PMID- 3051948 TI - [A case of cystic pheochromocytoma]. AB - A case of cystic pheochromocytoma is reported. A 64-year-old man was referred to our hospital because of right flank pain and gross hematuria. Excretory urography showed an inferior displacement of his right kidney, and right suprarenal mass was suspected. CT and ultrasonographic findings suggested that the most likely diagnosis was cystic adrenal tumor. Right adrenal tumor was removed through a transverse abdominal incision on October 15, 1986. During the procedure, the blood pressure rose to 260/100 mmHg at the time of tumor manipulation. The histological diagnosis was pseudocystic pheochromocytoma. Pathogenesis of cystic adrenal tumor is discussed briefly. PMID- 3051949 TI - [Myelolipoma of the adrenal gland: report of a case]. AB - A 50-year-old male was admitted to our municipal hospital because of his right suprarenal tumor which had been found by ultrasonography by chance at National Toneyama Hospital. His physical examination was normal except for obesity. Hematological examination and blood chemistry were normal and no endocrine disorder was found. Excretory urogram revealed neither deformity of the collecting system nor displacement of the right kidney. Computed tomography visualized sharp marginated in homogeneous structured mass of different densities within the negative range. Angiography revealed hypovascularity of the tumor. Right adrenalectomy was performed and the specimen weighed 72 g. Histopathologically is consisted of adult fat tissue and hematopoietic tissue. In addition to our case, 17 cases of surgically removed myelolipoma reported in the Japanese literature are reviewed. PMID- 3051950 TI - [Xanthogranulomatous pyelonephritis: clinical experience of 8 cases]. AB - Xanthogranulomatous pyelonephritis (XGP) is an uncommon form of granulomatous inflammation characterized by destruction of the renal parenchyma and replacement by solid sheets of lipid-laden macrophages. The first report was made by Schlagenhaufer in 1916. Due to diagnostic difficulties, relatively few cases have been reported. However, within the past 20 years, XGP has been recognized with increasing frequency. At present, over 400 cases have been reported. Recently, the computed tomographic (CT) findings of XGP have been described in a few sporadic case reports (1-4). Our clinical experience consists of 8 cases of XGP. The sonographic and computed tomographic findings in our cases are presented along with the correlation with the pathological specimens. We stress the importance of sonography and CT in preoperative planning. Moreover, in our cases we could obtain typical findings on sonography and CT, which enabled us to make a correct preoperative diagnosis. In this report we describe some specific features. PMID- 3051951 TI - [A case of Mondor's disease of the penis]. AB - A case of Mondor's disease of the penis in a 40-year-old man is reported. The patient complained of a small subcutaneous induration (0.5 x 1.0 cm) with slight tenderness in the dorsal region of the penile shaft. On examination, the linear cord was palpated running both distally and proximally from the induration. This lesion was removed under local anesthesia, and the induration and the cord were found to be part of the superficial dorsal vein of the penis. The venous wall was thick and the thrombus was packed in it. Histological findings showed the proliferation of connective tissue of the vessel wall and partially granulating thrombus in the canal. From these findings, we confirmed the diagnosis of Mondor's disease of the penis. The etiology of this disease, especially in comparison with non-venereal sclerosing lymphangitis of the penis (N.S.L.P.) is discussed. PMID- 3051952 TI - [A case of bilateral spermatocele]. AB - A case of bilateral spermatocele is reported. A 46-year-old man was admitted to our hospital complaining of bilateral scrotal swelling. We obtained a colorless, opalescent fluid which contained numerous spermatozoa. The fluid volume, pH and gravity obtained from right spermatocele were 85 ml, 6.8 and 1.005, respectively, and those obtained from left side were 40 ml, 6.8 and 1.006, respectively. Spermatocelectomy was done under lumbal anesthesia. Both spematocele were found near the body of the epididymis. The wall of spermatocele had on epithelial lining of cuboidal cells. Twenty two cases of spermatocele reported in Japan since 1951 are reviewed. PMID- 3051953 TI - [Primary carcinoid tumor of the testis: a case report]. AB - A case of pure, primary testicular carcinoid tumor in a 27-seven-year-old male is reported. The patient presented with a painless enlargement of the right testis but the serum markers for testicular cancer, including alpha-fetoprotein, beta human chorionic gonadotropin and lactate dehydrogenase, were not elevated. Right orchiectomy was performed. Histologically, the tumor showed a typical appearance of carcinoid tumor. Further examinations such as barium studies, computed tomographic scan and Ga scintigraphy, showed no other lesions, and he received an adjuvant chemotherapy of cyclophosphamide, 5-fluorouracil and adriamycin. He is well and free from symptoms 17 months after surgery. PMID- 3051954 TI - [Epidermoid cyst of the testis: a report of 2 cases]. AB - We report two cases of intratesticular epidermoid cyst, one of which had ossification in the cyst. Two cases were treated by orchiectomy with high ligation of the cord under the diagnosis of testicular malignant tumor. Ultrasonographic examination revealed a well-defined solid mass with echogenic rim. The internal echo of the cysts were relatively homogeneous and almost similar to the surrounding normal testicular parenchyma in the echo level. PMID- 3051955 TI - [Clinical studies on enoxacin in urinary tract infection]. AB - Enoxacin was administered orally to 49 out-patients with urinary tract infections (UTI) at the doses of 600 mg/day and clinical effects were evaluated. Out of 13 patients with simple acute UTI evaluated by the UTI criteria, the results were excellent in 8 cases and good in 5 cases. Overall effectiveness rate was 100%. Out of 10 patients with simple acute UTI evaluated on over 5-day administration, the results were excellent in 9 cases and good in 1 case. Overall effectiveness rate was 100%. Out of 14 patients with simple acute UTI evaluated by our own criteria because their bacteriological response is unknown, the results were excellent in 7 cases and good in 7 cases. Overall effectiveness rate was 100%. Out of 12 patients with chronic complicated UTI evaluated by the UTI criteria, the results were excellent in 6 cases, good in 3 cases and poor in 3 cases. Overall effectiveness rate was 75.0%. Poor effective cases were mainly found in ones with mixed infection. Sensitivity test using discs did not always correspond to the bacteriological effect in cases with chronic complicated UTI. An adverse reaction was noted in 3 patients (6.1%). Two cases had gastrointestinal symptoms and 1 case had skin eruption. All symptoms were readily improved after discontinuing administration. From the above results, Enoxacin was considered to be a useful agent for urinary tract infection. PMID- 3051956 TI - Diagnostic criteria for carotid duplex sonography. AB - Optimal criteria for the duplex sonographic diagnosis of carotid artery stenosis have not yet been defined. We studied 205 vessels in 105 patients with both duplex sonography and angiography. Four diagnostic groups were defined on the basis of Doppler flow characteristics. Receiver-operating-characteristic curves were used to compare diagnostic criteria at significant stenosis levels, and to select threshold values that emphasize specificity as well as sensitivity. Peak systolic velocity, systolic velocity ratios, and end-diastolic velocity were all shown to be equivalent predictors of significant disease. We chose peak systolic velocity in the internal carotid artery as our best parameter because of its ease of measurement. The use of combined parameters offered no significant statistical advantage over the use of a single parameter. The real-time assessment of stenosis and ulceration was not found to be reliable. ROC curves should be used to select Doppler criteria with desired sensitivities and specificities to maximize the benefit in each clinical setting. PMID- 3051957 TI - Pseudoaneurysm of the vertebral artery: appearance on color-flow Doppler sonography. PMID- 3051958 TI - The usefulness of diagnostic tests. PMID- 3051959 TI - Bioeffects of sonography. PMID- 3051960 TI - Myocardial paramagnetic contrast agents for MR imaging. AB - Several different paramagnetic contrast agents have been investigated for use in myocardial MR imaging. Gd-DTPA, the most extensively studied agent, has been shown to improve the conspicuity of acute myocardial infarcts on MR images in experimental animals and humans. However, this agent is limited as a marker of perfusion because of its rapid elimination by renal excretion and equilibration within the extracellular fluid space. Future investigation of Gd-DTPA as a myocardial perfusion agent may involve rapid-scanning techniques to define time dependent accumulation of the contrast agent in normal and ischemic myocardium during the first pass after IV injection. Nondiffusable paramagnetic agents and agents with prolonged retention in myocardium are being studied actively, but further tests of toxicity and metabolism are needed before clinical trials. Additional macromolecular-bound metal chelates will be tested in the future. It is hoped that these agents will allow detection of the jeopardized region of myocardium in the setting of acute ischemia, before the onset of myocardial edema. PMID- 3051961 TI - The role of sonography and transhepatic cholangiography in the diagnosis of biliary complications after liver transplantation. AB - We retrospectively reviewed the results of real-time sonography in 41 patients in whom biliary complications after liver transplantation were documented by percutaneous transhepatic cholangiography. Abnormalities included bile duct stricture (26 cases), occluded internal biliary stent (six cases), common duct redundancy with resultant functional biliary obstruction (three cases), bile leak (three cases), choledocholithiasis (two cases), and an abscess in a cystic duct remnant (one case). Sonography was abnormal in 22 of the 41 cases (sensitivity, 54%). Bile duct dilatation was the positive sonographic finding in 19 (86%) of the 22 abnormal examinations. In the remaining 19 patients, sonography was normal. Sonography is not a reliable test for the early detection of biliary abnormalities after liver transplantation. Percutaneous transhepatic cholangiography should be performed in patients with suspected biliary complications after liver transplantation. PMID- 3051962 TI - Comparison of a new colon lavage solution (Golytely-RSS) with a standard preparation for air-contrast barium enema. AB - Golytely is a balanced electrolyte solution for per oral whole-gut lavage. It has been an accepted alternative to standard preparations for barium enema. Golytely RSS is a new formulation designed to taste better through reduction in sodium and sulfate content. In addition, a slight increase in osmolality is intended to decrease further the net absorption/secretion ratio. One hundred outpatients who were to undergo air-contrast barium enema examinations were randomly assigned one of two colon-cleansing preparations. A standard 1-day diet/cathartic method was compared with Golytely-RSS plus oral bisacodyl. Radiographs were evaluated for the amount of fecal material present and the degree of mucosal coating by two radiologists independently of each other and without knowledge of the preparation used. Good to excellent feces removal was provided by Golytely-RSS in 98% of patients and by the standard method in 95% (p greater than .1, not significant). Mucosal coating also was similar between groups. Unpleasant side effects were reported significantly more often in the lavage group. Unwillingness to use the preparation a second time was reported in 36% of the Golytely-RSS group and in 8% of the standard preparation group (p = .03). Golytely-RSS is an acceptable alternative method to standard colon-cleansing methods for air-contrast barium enema and may be preferable in patients with certain renal or cardiovascular diseases. Side effects may preclude the use of this colon-cleansing regimen routinely. PMID- 3051963 TI - Sonography of the prostate: in vitro correlation of sonographic and anatomic findings in normal glands. PMID- 3051964 TI - Myelolipoma of the adrenal gland: sonographic and CT features. AB - The radiologic findings in 13 patients with pathologically proved adrenal myelolipomas were reviewed. All lesions involved the right adrenal gland; they ranged from 2 to 9 cm in diameter. Sonograms showed hyperechoic tumors in 11 cases (five homogeneous, six heterogeneous). Two myelolipomas composed primarily of myeloid tissue were hypoechoic. A propagation speed artifact was seen in seven lesions, all of which were composed primarily of fat and larger than 4 cm. CT identified fat-density tissues in all lesions. Contrast-enhanced CT showed positive attenuation values in the two predominantly myeloid tumors. CT appears to be sensitive for the diagnosis of adrenal myelolipomas. However, precontrast images are required to avoid errors. PMID- 3051966 TI - Fetal aortic and pulmonary artery diameters: sonographic measurements in growth retarded fetuses. AB - The small size of the head and abdomen in a growth-retarded fetus may lead to inaccurate estimates of gestational age. Therefore, we conducted a study to assess the diameters of the aortic root and pulmonary artery in such fetuses. During the study period, we measured these great vessel diameters at the level of the semilunar valves on all third-trimester obstetric sonograms in which intrauterine growth retardation was suspected. Cases were included in the study if delivery occurred within 1 week of the scan and if growth retardation was confirmed after birth. The study population consisted of 75 growth-retarded fetuses. We compared the relationships between great vessel diameters and gestational age in this study population with norms previously established in our laboratory in 403 normal fetuses. In a similar fashion, we compared the relationships between great vessel diameters and biparietal diameter in our study population with previously established norms. We found no significant difference between great vessel diameters in growth-retarded fetuses and those in normal fetuses of the same gestational age (p greater than .45 for aorta; p greater than .40 for pulmonary artery). In contrast, aortic root diameter was larger in relation to biparietal diameter in growth-retarded fetuses than it was in normal fetuses (p less than .05), and there was a tendency toward the same result for the pulmonary artery. We conclude that the diameters of the aorta and pulmonary artery remain normal in most cases of intrauterine growth retardation. The aorta and pulmonary artery may be useful predictors of gestational age when growth retardation is suspected. PMID- 3051965 TI - Sonographic evaluation of the fetal stomach: significance of nonvisualization. AB - The stomach was successfully visualized in 1051 (98%) of 1071 consecutive sonograms obtained in 995 fetuses after 14 weeks gestational age. All patients were studied prospectively. Stomach nonvisualization was associated with an abnormal outcome in 55% of the fetuses studied after 14 weeks and in 100% of the fetuses studied after 19 weeks. Fetal abnormalities included gastrointestinal and CNS malformations. Oligohydramnios was often present. The absence of a stomach on fetal sonograms obtained after 14 weeks gestational age strongly suggests fetal abnormality. Repeat sonograms should be obtained in all such cases. PMID- 3051967 TI - Percutaneous nonendoscopic gastrostomy in children. AB - Surgical gastrostomy has long been a standard method of providing nutrition to infants and children. Recently, percutaneous endoscopic gastrostomy has been advocated as a safer, quicker, less expensive method in children. We report our experience with 16 percutaneous gastrostomies in 14 infants and children; in all cases, both sonographic and fluoroscopic guidance were used. Four patients had had previous surgical gastrostomy in which the tubes could not be replaced once they were removed. The remaining patients were referred for percutaneous placement of gastrostomy tubes as the first procedure. In 13 procedures, parenteral sedation and local anesthesia were used; the remaining three procedures were done with the patient under general anesthesia. Tubes were successfully placed in all procedures. In two patients, tubes became dislodged, necessitating a second procedure. There were no instances of local infections, hemorrhage, or peritonitis, and none of the patients died. Two patients had postprocedure septicemia, which responded to antibiotics. Percutaneous nonendoscopic gastrostomy can be safely and effectively performed in infants and children. PMID- 3051968 TI - Massachusetts: omen or anomaly? PMID- 3051970 TI - New antibiotics: carbapenems, monobactams and quinolones. AB - New beta lactams and the quinolone class of antibiotics represent major improvements in the therapy of moderate to severe infections. These newer antibiotics have an extended spectrum of antimicrobial activity, excellent pharmacokinetic properties and low toxicity. The beta lactams include carbapenems, represented by imipenem-cilastatin, and monobactams, represented by aztreonam. Norfloxacin and ciprofloxacin are potent quinolones. PMID- 3051971 TI - Selecting a screening mammography facility. AB - Breast cancer is the second most common cause of cancer mortality in women. During the past 20 years, two major mammography screening programs have demonstrated reduced mortality in patients over age 50. Mammography can be safe and effective. Physicians should recommend mammographic facilities with trained and experienced personnel, dedicated mammographic and processing equipment, and quality control procedures. PMID- 3051969 TI - The infertile couple. AB - The causes of infertility can be divided into seven categories. In older couples, valuable time can be lost if evaluation is delayed. An orderly diagnostic approach assures that no diagnosis is overlooked. Many of the causes of infertility are amenable to therapy. PMID- 3051972 TI - Newborn circumcision. AB - Routine circumcision of male newborns continues to be practiced widely in the United States, although many physicians believe there is little medical indication for the procedure. The parents' decision regarding circumcision is often based on inadequate information; however, attempts to provide information to parents have not had much impact on the frequency of circumcision. Many third party payers have begun to refuse payment for the procedure, and there are indications that this action will diminish the number of circumcisions. PMID- 3051973 TI - Scalp psoriasis. AB - Many people suffer from scalp psoriasis. While the cause of this condition is not known, there is effective treatment. With well-planned therapy and patient compliance, scalp lesions can be controlled. Useful agents include coal tar shampoos and preparations, keratolytics and topical corticosteroids. Systemic agents are almost never indicated for scalp psoriasis alone. PMID- 3051974 TI - Seasonal depression. AB - Seasonal affective disorders are mood disturbances that occur with a change in season. The most common form is winter depression, marked by sadness, anxiety, decreased physical activity, increased appetite, carbohydrate craving, weight gain, hypersomnia, decreased libido, worsening of premenstrual symptoms, impaired work performance and interpersonal conflict. This syndrome often responds to daily exposure to bright artificial light. PMID- 3051975 TI - Cyanosis due to pulmonary arteriovenous malformation. AB - Cyanosis is frequently encountered in the neonate or infant. Most often it is due to congenital heart disease or primary lung disease. Pulmonary arteriovenous malformation is an unusual cause of cyanosis. Polycythemia and clubbing are associated findings. The treatment is excision or embolization if the disease is not too extensive. PMID- 3051976 TI - Use of stimulant medications in children. AB - A systematic approach to the use of stimulant medications is required in the treatment of attention deficit disorder and associated learning problems. The diagnosis of attention deficit disorder must be confirmed before drug therapy is started, and parents must be counseled about the use of stimulant medications. The actions and side effects of the various treatment alternatives should be discussed. PMID- 3051977 TI - Tuberculosis update. AB - Tuberculosis cases are increasing in the United States, partly because of the emergence of AIDS and the rise in the homeless population. Faster culture methods usually allow identification within two weeks. Research is being done on serologic and recombinant nucleic acid methods of detecting tuberculosis in various body fluids and secretions. A six-month treatment regimen for tuberculosis has proved to be effective, as have shorter courses of prophylactic therapy for tuberculin skin test converters. PMID- 3051978 TI - Acute diarrhea. AB - Acute diarrhea can be life-threatening in the very young, the elderly and the malnourished. Osmotic diarrhea is produced by unabsorbed solutes in the intestinal lumen, while exudative diarrhea is caused by infection and inflammation. Secretory diarrhea results from enterotoxins, oversecretion of gastrointestinal hormones and the action of bile acids. Rapid intestinal transit also may cause diarrhea. PMID- 3051979 TI - Health hazards of farming. AB - The U.S. farm work force numbers approximately 6.5 million. The health risks connected with their farm tasks are many and varied. Accidents and illnesses are caused by tractors, specialized farming equipment, chemicals, zoonoses, fungi, sensitizers, and the environment of confinement buildings. The scattered nature of the work force makes preventive measures difficult to implement. PMID- 3051981 TI - Atrial infarction: diagnosis and management. AB - Atrial infarction has been a relatively understudied entity. Its incidence by autopsy study has been widely variable, from 0.7% to 42%, with the largest series of 182 patients demonstrating an incidence of 17%. The right atrium is involved five times as often as the left, with the auricle the predominant site in either atria. Clinical atrial infarction may present with supraventricular arrhythmias, atrial rupture, hemodynamic compromise from loss of atrial "kick," and thromboembolic phenomena. Diagnosis currently is made in an appropriate clinical setting with characteristic PR interval changes. Other noninvasive techniques have shown only limited diagnostic utility, but esophageal echocardiography may prove to be a useful technique in this setting. PMID- 3051980 TI - Diagnosis and staging of HIV infection. PMID- 3051982 TI - How effective is digitalis in the treatment of congestive heart failure? AB - The evidence suggests that digitalis glycosides do indeed improve ventricular performance through a sustained but moderate positive inotropic effect. This effect is more marked in failing than in nonfailing myocardium. The clinical studies suggest a moderate salutary effect in patients with chronic CHF who are in sinus rhythm. The drug can be given safely to patients with CAD and in combination with other medications when the physician is aware of those factors leading to increased sensitivity to digitalis. PMID- 3051983 TI - Detection and treatment of acute pulmonary embolism: the use of a portable video image processor. PMID- 3051984 TI - Ambulatory blood pressure monitoring: practical considerations. AB - Ambulatory blood pressure monitoring (ABPM) allows one to evaluate the blood pressure (BP) profile over a 24-hour period in the patient's natural environment. Casual pressure measurements in the physician's office can be affected by alarm reactions, thus causing "white coat" hypertension. ABPM allows one to evaluate these reactions and determine the average pressure and variability of BP along with the effects of physical activity and emotional arousal on BP patterns while at work, at home, and during sleep. Average pressures determined by ABPM are more predictive of target organ involvement and cardiovascular complications of hypertension than casual monitoring of BP in the clinic. The absence of physiologic decline in arterial pressure during sleep is associated with increased prevalence of atherosclerotic complications and left ventricular hypertrophy as well as impairment of the autonomic nervous system. Although further prospective studies are needed to confirm the benefits of home pressure readings and ABPM, ABPM can be helpful in the diagnosis and determination of prognosis and therapeutic responses in a select group of patients. PMID- 3051985 TI - Effect of reperfusion on electrocardiographic and enzymatic infarct size: results of a randomized multicenter study of intravenous anisoylated plasminogen streptokinase activator complex (APSAC) versus intracoronary streptokinase in acute myocardial infarction. AB - The effect of early coronary artery reperfusion on ECG and enzymatic parameters was examined in 240 patients with acute myocardial infarction. These patients had participated in a randomized trial comparing intravenous anisoylated plasminogen streptokinase activator complex (APSAC) (n = 123) and intracoronary streptokinase (n = 117) therapy. Reperfusion occurred in 59 of 115 (51%) patients receiving APSAC and 67 of 111 (60%) patients receiving streptokinase (p = NS). There was greater early resolution of ST segment elevation in the reperfused than in the nonreperfused patients (p less than or equal to 0.003) and more rapid Q wave evolution (p less than or equal to 0.03). Sigma Q was lower in reperfused than in nonreperfused patients at 8 hours (1.41 +/- 1.18 versus 2.11 +/- 2.10 mV; p less than or equal to 0.05) and at 24 hours (1.43 +/- 1.25 mV versus 2.08 +/- 1.88 mV; p less than or equal to 0.02). Time to peak level was shorter in the reperfused patients for creatine kinase (CK) (10.7 +/- 5.5 hours versus 14.9 +/- 5.9 hours; p less than 0.0001) and lactic acid dehydrogenase (LDH) (29.6 +/- 13.6 hours versus 34.4 +/- 10.5 hours; less than or equal to 0.03) enzymes. Peak LDH-1 was lower in the reperfused group (274 +/- 149 U/L versus 341 +/- 173 U/L; p less than or equal to 0.04). Reperfusion at a mean of 3.9 hours after the onset of infarction was associated with more rapid resolution of ST segment elevation, faster Q wave evolution, smaller ECG infarct size, earlier cardiac enzyme release, and smaller enzymatic infarct size than later or no reperfusion. PMID- 3051987 TI - 'The personal growth opportunities that we can create in the VA will help in the recruitment and retention of pharmacists'. PMID- 3051986 TI - Steroid-induced enhancement of functional recovery of postischemic, reperfused myocardium in conscious dogs. AB - The effects of methylprednisolone sodium succinate (20 mg/kg, intravenously administered) on the time course of functional recovery of myocardium following a 15-minute coronary artery occlusion period and subsequent 5 hour reperfusion period were studied in chronically instrumented, conscious dogs. In comparison to a control group, animals receiving methylprednisolone 90 minutes prior to coronary occlusion demonstrated less depression of regional segment shortening following 15 minutes of reperfusion (52 +/- 13% vs control levels of 23 +/- 7% of preocclusion values) and improved recovery at 5 hours postreperfusion (106 +/- 6% vs control levels of 54 +/- 4% of preocclusion values). In animals receiving methylprednisolone immediately prior to reperfusion, there was also similar recovery of segment shortening at 5 hours (97 +/- 3%). In contrast, dogs receiving methylprednisolone 15 minutes after the onset of reperfusion or sodium succinate (5.5 mg/kg, intravenously administered) 90 minutes prior to occlusion demonstrated no improvement in recovery of function. Experiments in dogs not subjected to coronary occlusion documented that methylprednisolone sodium succinate lacked inotropic and vasodilator properties. The results suggest that methylprednisolone administered prior to or during coronary artery occlusion but not after reperfusion enhances the functional recovery of hypokinetic, postischemic, reperfused myocardium. These effects are unrelated to any direct hemodynamic action of steroids or to the sodium succinate salt. PMID- 3051988 TI - Current ideologies of eating disorders: an overview. PMID- 3051989 TI - Clinical laboratories for the practicing pharmacist: general serum chemistry. PMID- 3051991 TI - Metabolic effects of antihypertensive therapy with a calcium antagonist. AB - The effect of diuretics to increase serum glucose, low-density lipoprotein cholesterol and triglycerides, as well as the adverse changes in triglycerides and high-density lipoprotein cholesterol produced by nonselective beta blockers, have been largely ignored in the treatment of hypertension. However, a number of trials have shown that reductions in serum lipids can alter cardiovascular mortality. Calcium antagonists have become major drugs in the treatment of hypertension, and some data suggest that calcium antagonists may increase serum glucose levels. Significantly less data on lipid effects have been published. Lipid and glucose effects were examined in an 8-week antihypertensive study using a sustained-release preparation of diltiazem titrated from 240 to 360 mg/day in a twice-daily regimen in a randomized, double-blind, placebo-controlled parallel trial in 96 patients. Average supine blood pressure at week 8 was 156/98 mm Hg, standing blood pressure with placebo 152/100 mm Hg, and with diltiazem 147/91 and 144/93 mm Hg. There were no statistically significant changes in serum lipids or glucose in the diltiazem or placebo group or between the groups. Mean values (mg/dl) at baseline and week 8 in the diltiazem group were, respectively, for cholesterol 215 and 218, high-density lipoprotein cholesterol 50 and 51, low density lipoprotein cholesterol 128 and 133, triglycerides 169 and 175, and glucose 113 and 110. Thus, this large and placebo-controlled study shows that diltiazem is among the antihypertensives with no adverse long-term lipid or glucose effects. PMID- 3051990 TI - Diltiazem and captopril alone or in combination for treatment of mild to moderate systemic hypertension. AB - The efficacy and safety of sustained-release diltiazem, 60 to 180 mg twice daily, was compared with that of captopril, 25 to 75 mg twice daily, alone and in combination, in 132 patients with mild to moderate essential hypertension (supine diastolic blood pressure [BP] 95 to 114 mm Hg). All patients received placebo for 4 to 6 weeks, followed by randomization to diltiazem or captopril during the double-blind monotherapy phase. Either study drug was titrated over 6 weeks to achieve a goal supine diastolic BP reduction of at least 10 mm Hg and a diastolic BP of less than 90 mm Hg. Patients achieving the goal BP reduction were maintained on monotherapy for an additional 8 weeks. Patients not achieving the treatment goal after 8 weeks with either drug alone received the other drug in combination, titrated to achieve goal BP response. Both drugs lowered BP significantly and, at the doses used, diltiazem had a greater effect on diastolic BP than did captopril. The mean changes from baseline at week 8 were -10.6 and 7.3 mm Hg, respectively, (p = 0.01). Goal BP was achieved in 38% of patients taking diltiazem monotherapy and in 34% of patients taking captopril monotherapy. There were no significant differences between diltiazem and captopril in diastolic or systolic BP reductions by race or age. The addition of alternate therapy for non-goal achievers at week 8 resulted in significant reductions in diastolic and systolic BP by week 16.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3051992 TI - Catecholamines, calcium and cardiomyopathy. AB - The cardiomyopathic Syrian hamster has a genetically transmitted form of dilated cardiomyopathy and is an important paradigm of myocardial disease, particularly for studies addressing the earliest stages of myocardial dysfunction. This model exhibits an increase in cardiac sympathetic tone in the presence of an altered expression of sarcolemmal calcium channels or of alpha 1 receptors, and a defective handling of calcium by both cardiomyocytes and vascular smooth muscle cells. Increased expression of the oncogene c-myc is evident in cardiomyocytes before any overt evidence of heart disease. Alterations in a nuclear phosphoprotein, which appears to be important in the regulation of gene expression, have also been identified. The disease becomes phenotypically manifest by the development of microvascular spasm, reperfusion injury and myocyte loss. Myocyte loss, in turn, burdens the remaining cells with an increasing load, increasing sympathetic stimulation, myocyte hypertrophy and further cell loss--a continuing vicious spiral that culminates in the development of myocardial failure. All of the features of hamster cardiomyopathy may be prevented by the administration of verapamil or prazosin to juvenile hamsters before the phenotypic onset of their heart disease. This understanding has led to the study of new imaging agents that promise the detection of such forms of cardiomyopathy in their earliest stages and a means by which the effects of therapy can be assessed. If such mechanisms are applicable to human cardiomyopathy, early treatment of patients with adrenergic antagonists or calcium antagonists should be beneficial. PMID- 3051993 TI - Cell biology of vascular hypertrophy in systemic hypertension. AB - Recent data demonstrate that in addition to its conduit function, the blood vessel is an active synthetic and secretory organ containing several autocrine and paracrine systems that are involved with the local regulation of its own function (i.e., structure and growth). The endothelium secretes vasorelaxant and vasoconstrictive substances, growth factors and inflammatory mediators that exert paracrine influences on vascular myocyte function. The vascular myocyte also expresses autocrine substances that influence its own function. The autocrine systems include angiotensin, prostaglandins, platelet-derived growth factor, insulin-like growth factor and heparin. These local factors exert modulatory influences on myocyte contractility and growth. These autocrine and paracrine systems serve as an adaptive mechanism by which the vasculature autoregulates its structural and functional state. We speculate that an alteration in this delicate balance of these local factors, due to genetic or acquired abnormalities, can result in increased vascular tone and vessel hypertrophy and thereby contribute to the pathogenesis of hypertension. PMID- 3051994 TI - Effects of calcium on renin and aldosterone. AB - A series of experiments was undertaken to assess the effects of calcium administration, in vivo, on renin and aldosterone secretion. In the anesthetized dog, renin secretion was decreased by renal arterial infusions of calcium chloride and calcium gluconate; aldosterone excretion was not affected. In the sodium chloride-deprived rat, dietary calcium chloride loading decreased plasma renin activity, whereas calcium gluconate did not. Both calcium salts increased aldosterone production. In the non-filtering, denervated, papaverine-treated dog kidney, renin release was stimulated by renal arterial infusion of verapamil. In the rat, chronic oral verapamil administration decreased plasma aldosterone but had no effect on renin. In humans, chronic oral verapamil decreased aldosterone responsiveness to infusion of angiotensin II. Thus, in vivo renin release is inhibited by hypercalcemia and stimulated by blocking calcium transport; conversely, aldosterone production is stimulated by a high calcium intake and inhibited by blocking calcium transport. These effects of calcium on renin and aldosterone may have implications for understanding the putative relation between calcium and hypertension. PMID- 3051995 TI - Mechanisms of the vascular effect of pressor hormones. AB - There exists a considerable gap between the recognition of the effects of pressor hormones and the understanding of their mechanisms. Studies are necessary to provide a more accurate basis for potential therapeutic interventions on the hormonal mechanisms of hyper- and hypotensive states. From recent and previous experimental results, a modified model of vascular smooth muscle cell activation is emerging. In this model, contraction is a complex process involving Ca2+ release, protein kinase C and ionic channel activation and modification in pH and energy metabolism. The combined action of these processes is necessary for the full expression of the contractile response. PMID- 3051996 TI - Mechanisms of hypertensive glomerular injury. AB - Systemic hypertension complicates the course of most patients with chronic renal failure and accelerates the progression of experimental and clinical glomerular disease. Based on recent experimental studies, it is suggested that at similar levels of systemic hypertension, glomerular injury only develops when pre glomerular resistances are ineffective, thus allowing the development of glomerular hypertension. The mechanisms by which the hemodynamic stress of elevated intracapillary flows and pressures leads to progressive glomerular damage, particularly to the development of focal glomerulosclerosis is currently unknown. Endothelial cell injury, increased mesangial traffic or trapping of macromolecules and epithelial cell injury, or a combination, appear to occur early, followed by in situ inflammatory and microthrombotic mechanisms. This may explain why various therapeutic approaches, whether dietary or pharmacologic, can have salutory effects despite their diverse mechanisms of action. PMID- 3051997 TI - Comparison of the effects of calcium antagonists and converting enzyme inhibitors on renal function under normal and hypertensive conditions. AB - Calcium antagonists decrease the ability of the kidney to autoregulate renal blood flow (RBF) and glomerular filtration rate (GFR). Therefore, when afferent renovascular resistance is elevated, as in essential hypertension, there is a resultant increase in RBF and GFR with the administration of calcium antagonists. These agents also induce a marked natriuresis because of direct tubular action through unknown mechanisms. The natriuresis can be dissociated from renal and systemic hemodynamic actions, indicating that the decreased sodium reabsorption could override other compensatory mechanisms explaining the absence of sodium retention during the treatment. The renal effects of converting enzyme inhibitors (CEIs) can be explained by the reduction of intrarenal formation in angiotensin II. Because the activation of the renin-angiotensin system is mainly responsible for inducing sodium retention during a decrease in systemic blood pressure, CEIs could have a protecting effect without disturbing other homeostatic mechanisms. CEIs decrease efferent glomerular resistance, reducing capillary pressure and thereby reducing GFR. This effect is not translated in sodium retention because the reduction of GFR is mild during captopril administration in kidneys with normal or increased renal perfusion pressure. At low renal perfusion pressure, the reduced glomerular afferent vasoconstriction can compromise GFR, leading to renal insufficiency. Although these situations are not likely to be encountered during the treatment of uncomplicated essential hypertension, in severe hypertension with hypertrophy of pre-glomerular vessels, glomerular perfusion may decrease. Combination therapy of calcium antagonists and CEIs has been reported to be an effective treatment of severe hypertension. Currently, little information is available on the manner in which renal function is affected by simultaneous administration of both drugs. PMID- 3051998 TI - Renal hemodynamic effects of a selected calcium antagonist. AB - Calcium antagonists are unique antihypertensive drugs that appear to exert selective blood pressure-lowering and possibly renal hemodynamic and functional effects in hypertensive patients and animals. There is evidence for inhibition of tubular sodium reabsorption and renal vasodilatation when certain of these agents are given by acute intravenous or intrarenal arterial administration. These renal effects have been observed to occur either independently or together. Both natriuresis and diuresis have been found to occur with these drugs. In the deoxycorticosterone-salt hypertensive dog, chronically administered diltiazem reduces blood pressure, transiently increases renal blood flow and increases urine volume. Administered either acutely or chronically to these hypertensive dogs, diltiazem depresses renal vascular reactivity. Pressor and renal vasoconstrictor responses to angiotensin II and norepinephrine are attenuated to a similar degree. The chronic blood pressure-lowering effect of diltiazem is most likely a function of depressed vascular reactivity; however, actions at other sites cannot be ruled out based on our experiments. Postprandial renal vasodilatation readily occurs in the conscious instrumented dog, and although this response is blocked by the acute administration of a calcium antagonist, the response is unaltered during the chronic administration of diltiazem. PMID- 3051999 TI - Emerging issues in the cellular biology of the cardiovascular system. AB - Current theory suggests that life began in a prebiologic era, progressed to a ribonucleic-deoxyribonucleic cellular era, and finally entered an era characterized by multicellular organisms. If this progression is correct, it is not surprising that, as medicine studies living organisms with increasing sophistication, factors that are initially discovered to have systemic effects are, in many instances, later determined to have paracrine, autocrine or even intracellular ("intracrine") effects. This schema is potentially of value in analyzing the pathogenesis of cardiovascular disease and, in particular, the development of the sequelae of hypertension. A case is made for the idea that the actions of common peptide and nonpeptide factors at local tissue levels can play an important role in the development of atherosclerosis and left ventricular hypertrophy. In making this case, the potential roles of insulin, angiotensin II and other vasoactive factors are considered. In addition, it is argued that some peptide and nonpeptide factors with cardiovascular impact may operate in the intracellular environment, thus broadening prospects for study and intervention. Finally, genomic alterations either spontaneously occurring or resulting from chronic stimulation or viral infection are considered and their potential role is discussed. PMID- 3052000 TI - Function of the hypertensive kidney during calcium flux manipulation. AB - Dihydropyridine calcium channel agonists and antagonists elicit exaggerated glomerular and circulatory responses from kidneys isolated from Dahl rats genetically programmed to develop NaCl-induced hypertension (Dahl S rats). These differential responses are further magnified by NaCl loading. In contrast, "chemical sympathectomy" with 6-hydroxydopamine enhances renal vascular responses to calcium channel agonists in a manner that depends on the antecedent dietary NaCl intake, and is independent of genetic predilection to develop NaCl-induced hypertension. These findings are consistent with the hypothesis that aberrations of vascular and perhaps glomerular calcium entry modulation may be determinants of altered renal hemodynamics in NaCl-sensitive hypertension. The latter may be responsible for the enhanced responsiveness to calcium channel antagonists observed in NaCl-sensitive hypertension in humans. PMID- 3052001 TI - Effects of calcium antagonists on deranged modulation of the renal function curve in salt-sensitive patients with essential hypertension. AB - Two subsets of patients with essential hypertension have been identified based on their sensitivity to dietary sodium intake: the salt-sensitive and the salt resistant patients. The renal function curve in salt-resistant patients is shifted to higher blood pressure levels but it remains parallel to that of normal subjects. On the contrary, the slope of the renal function curve in salt sensitive patients is considerably depressed. It is postulated that this derangement may be related to an abnormal relation between sodium intake and sympathetic nervous system activity. It is also postulated that a link exists between abnormalities of sodium and calcium metabolism in hypertension and that reduced renin release, increased sympathetic activity and reduced renal sodium excretion in salt-sensitive patients may be related to a defect in sodium-linked cellular calcium transport. Calcium antagonists revert the derangements in the renal function curve and in renin release in salt-sensitive patients. PMID- 3052002 TI - Determinants of antihypertensive response to calcium antagonists in systemic hypertension. AB - Calcium antagonists are an important group of drugs in the treatment of systemic hypertension. Examination of the factors that determine the magnitude of antihypertensive response to calcium antagonists is of interest for clinical and basic pathophysiologic reasons. Possible factors influencing response include drug dose, plasma drug concentration, plasma renin activity, baseline blood pressure, nonionized calcium in blood and inside vascular smooth muscle cells, patient age and patient race. Published studies have examined the determinants of antihypertensive response. The striking fact about many of these studies is that they fail to control for other possible factors that might influence response. This article reviews the methods used and the results of 23 different studies that examined the determinants of response to calcium antagonists. Possible relationships between plasma renin activity, intra- and extracellular calcium levels, and the antihypertensive response, are of interest from a pathophysiologic point of view. From a clinician's standpoint, the only determinants that matter are the dose of the drug and the degree of elevation of the blood pressure. In this regard, calcium channel blocking drugs are no different from other drugs that act by peripheral vasodilation. PMID- 3052003 TI - Automated blood pressure monitoring for the assessment of antihypertensive treatment. AB - Whole-day automated monitoring with small portable devices allows the circadian pattern of blood pressure to be assessed conveniently in individual patients. Measurements provided by this technique appear to be more reproducible and physiologically relevant than conventional office readings. Ambulatory monitoring can enhance the evaluation of antihypertensive therapy, especially in clinical trials, in 3 ways. First, it can facilitate the diagnosis of hypertension and avoid potentially inappropriate treatment in patients whose hypertension is erroneously diagnosed by office measurements. Moreover, the monitoring technique appears to prevent placebo responses in therapeutic studies. Second, whole-day monitoring allows clear quantification of treatment effects. Because the standard deviations of treatment-induced blood pressure differences are lower with this method than with conventional readings, valid statistical evaluation of antihypertensive effects can be performed with far fewer patients. Finally, duration of action of drugs can be measured. By dividing the day into sequential periods (typically 12 two-hour periods), it is possible to determine statistical differences between baseline and treatment blood pressure values throughout the dosing intervals of the drugs being tested. This method may be more sensitive than the traditional "peak and trough" approach for assessing duration of action; it is not influenced by unsuspected discrepancies between the pharmacokinetic and pharmacodynamic properties of drugs, and can also provide information on nighttime effects. Thus, automated whole-day blood pressure monitoring appears to be a powerful tool for the evaluation of antihypertensive therapy. PMID- 3052004 TI - Evaluation of antiarrhythmic therapy using Holter monitoring. AB - Premature ventricular complexes and nonsustained ventricular tachycardia mark a person with structural cardiac disease as a high--risk candidate for sudden cardiac death. Such ventricular arrhythmias are considered potentially lethal and should be distinguished from both those that are benign and those that cause hemodynamic consequences (i.e., lethal or malignant arrhythmias). Noninvasive Holter monitoring is the principal technique for detecting and evaluating the presence of potentially lethal ventricular arrhythmias. These arrhythmias undergo a high degree of spontaneous variability. Thus, to define a therapeutic drug effect, a reduction in the frequency of premature ventricular complexes of at least 75% and a reduction in the frequency of nonsustained ventricular tachycardia by at least 90% are required to eliminate the likelihood of spontaneous variability as the cause of this change in the frequency of arrhythmia. To define proarrhythmia, a different algorithm must be applied. When using antiarrhythmic drugs, a quantitative ventricular arrhythmia baseline for both frequency and type of arrhythmia must be established so that after therapeutic intervention repeat Holter monitoring can determine whether efficacy, inefficacy or proarrhythmia had occurred. Holter monitoring clearly reveals differential antiarrhythmic response rates among classes of antiarrhythmic drugs in patients with benign or potentially lethal arrhythmias. However, preliminary data have not clearly defined the relation between antiarrhythmic pharmacotherapy and a reduction in sudden cardiac death. The results of large-scale clinical trials that have only recently been undertaken must be assessed to determine whether sudden cardiac death can be prevented by adequately suppressing potentially lethal ventricular arrhythmias. PMID- 3052005 TI - Initial dosage selection of antiarrhythmic therapy. AB - The success or failure of antiarrhythmic drug treatment depends, in part, on the selection of the initial dosage. Too low a dosage can lead to unnecessary (and frequently life-endangering) delays in achievement of arrhythmia suppression. Conversely, an excessively high dosage can lead to intolerable toxicity and cessation of treatment. The recommended approach to therapy is to begin with a relatively low dosage, i.e., the lowest dosage with a reasonable chance of producing a favorable response, and titrating the dose upward as needed. Dose titration should be guided by clinical response and, when appropriate, concentrations of the drug and any active metabolites in the plasma. In situations frequently encountered in practice, however, the initial dosage must be modified because of interindividual differences in drug disposition. These changes in drug pharmacokinetics can arise from a variety of factors, including disease processes (e.g., congestive heart failure, cirrhosis and renal failure), concomitant medications (e.g., hepatic enzyme inducers such as phenytoin and inhibitors such as amiodarone), drug formulation, protein binding and inherited drug metabolism capacity. Knowledge of these factors can help the clinician to avoid potential pitfalls in initial dosage selection and can enhance the changes of successful drug treatment of arrhythmias. PMID- 3052006 TI - The role of exercise testing in evaluation of arrhythmias. AB - Exercise testing has been widely applied for the evaluation of patients with coronary artery disease. The principles underlying its use for this indication make it a useful adjunctive technique, when combined with ambulatory monitoring, to diagnose arrhythmias and monitor antiarrhythmic drug therapy. During exercise, there is a withdrawal of vagal tone and a marked increase in circulating catecholamines and sympathetic inputs to the heart. These changes may directly cause arrhythmias (e.g., catecholamines can enhance automaticity and delayed afterpotentials and can shorten myocardial conduction time and refractory periods). However, they also augment myocardial oxygen demands by increasing myocardial inotropy, heart rate and blood pressure. Such changes may cause ischemia in patients with heart disease, which is a powerful stimulus for arrhythmia, or lead to dysfunction in left ventricular contraction and increased myocardial wall stress, factors that also may precipitate arrhythmia. In approximately 10% of patients with a history of serious arrhythmia, exercise represents the only means for exposing arrhythmia. Importantly, this technique is useful for evaluating the effect of antiarrhythmic drugs. These agents work by reducing membrane automaticity, slowing impulse conduction through the myocardium and prolonging membrane refractoriness. In contrast, catecholamines, which are secreted in response to exercise, have the opposite effect. Thus, exercise may negate the important effects of the antiarrhythmic drugs. Additionally, exercise testing may expose potentially serious toxic drug reactions that may not be obvious at rest. These include conduction abnormalities, negative inotropic effects, congestive heart failure and aggravation of arrhythmia. Although the presence and frequency of arrhythmia with exercise is highly variable in patients with benign arrhythmia, results are more consistent in patients with a history of serious arrhythmia. If arrhythmia is reproducibly provoked with exercise, this technique can be used to judge drug effect. Thus, exercise testing is an important, reliable and helpful technique for exposing arrhythmia, evaluating drug efficacy and identifying potentially serious toxic drug effects. PMID- 3052007 TI - Role of the electrocardiogram in determining electrophysiologic end points of drug therapy. AB - Electrocardiographic monitoring can be a useful adjunct to antiarrhythmic therapy, since the electrocardiogram is a simple indicator of net cardiac drug effect, irrespective of factors such as pharmacokinetic variability, drug metabolite interactions or intraindividual variability in drug sensitivity. Changes associated with antiarrhythmic drug therapy include markers of sodium channel block, such as increased QRS or sinus-ectopic coupling intervals and increased QT interval, a marker of action potential prolongation. Electrocardiographic changes can serve 3 purposes: They can correlate with arrhythmia suppression, they may be a guide to impending drug toxicity, and they can indicate the presence of an antiarrhythmic drug at some electrophysiologically active site in the heart. This latter indication may be used as an assessment of compliance, as a clue to drug-drug interactions that may lower antiarrhythmic drug concentrations or preparatory to electrophysiologic testing when it is desirable to avoid testing patients who have no demonstrable drug effect. Drug-induced changes in the microelectrophysiologic environment may sometimes fail to express themselves on the surface electrocardiogram. Overall, however, the electrocardiogram is an inexpensive, readily available tool to monitor net antiarrhythmic drug effects on the heart. Monitoring of the electrocardiogram should, therefore, be an integral part of managing antiarrhythmic drug therapy in patients with arrhythmias. PMID- 3052008 TI - Electrophysiologic evaluation of antiarrhythmic therapy for ventricular tachyarrhythmias. AB - The use of electrophysiologic studies has contributed significantly to our understanding of the mechanisms of ventricular tachyarrhythmias and enhanced our ability to assess objectively the efficacy of various therapeutic interventions in modifying or preventing their recurrence. The basis on which electrophysiologic testing techniques is founded is the ability reproducibility to initiate ventricular arrhythmias by programmed electrical stimulation in patients with a history of recurrent ventricular tachycardia or fibrillation. Ventricular tachycardia can be initiated by electrophysiologic studies in approximately 90% of patients with clinically documented recurrent, sustained ventricular tachycardia related to coronary artery disease and in 60% of patients with nonsustained ventricular tachycardia. Reports indicate that electrophysiologic testing is highly specific as well (99% for sustained monomorphic ventricular tachycardia). Studies in patients with recurrent ventricular tachycardia demonstrate that prevention by antiarrhythmic drugs of the ability to initiate tachycardias that were previously inducible by comparable stimulation techniques in the absence of therapy is highly predictive of freedom from recurrent episodes of spontaneous ventricular tachycardia and ventricular fibrillation. This end point can be achieved in 35 to 75% of patients. This wide range of success rates results from differences in the patient populations studied, as well as major differences in the programmed stimulation and antiarrhythmic drug protocols used among laboratories. The positive predictive value of this technique (defined as the percentage of patients in whom complete suppression of inducible ventricular tachycardia or ventricular fibrillation is achieved during electrophysiologic testing with antiarrhythmic drugs and in whom no spontaneous arrhythmia occurs at 1- to 2-year follow-up) ranges between 80 and 95%. PMID- 3052009 TI - Role of plasma concentration monitoring in the evaluation of response to antiarrhythmic drugs. AB - Plasma concentration monitoring of antiarrhythmic agents is valuable, but it is often misused or overemphasized in therapeutic decision-making. There are strict requirements for its appropriate use that are often not met--for both the newer and even the conventional antiarrhythmic drugs. For maximum value, there must be a reliable, accurate relation between the plasma drug concentration and drug action, a relation closer than that between dosage and drug action. The time of sample collection is important--most guidelines are based on "trough" plasma concentrations measured after steady-state equilibrium has been achieved. The use of an accurate, sensitive and specific assay is crucial to the value of plasma concentration monitoring guidelines. However, for agents having active metabolites, monitoring the concentration of only the parent drug can be misleading and limits (but does not necessarily eliminate) the value of plasma concentration monitoring guidelines for these agents. Plasma concentration monitoring of most antiarrhythmic agents is of value for certain specific purposes: to determine compliance to antiarrhythmic therapy, to detect and analyze possible drug interactions, to assess the benefit to risk ratio for increasing the dose of a particular antiarrhythmic agent, to maintain a stable drug effect in the presence of a patient's changing clinical condition and, to a limited extent, to assess the role of an agent in causing an adverse drug reaction. The importance of understanding the assay methods currently in use, as well as how plasma concentration monitoring of individual antiarrhythmic agents is affected by the presence of active metabolites, optical isomers differing in their activity and variations in protein binding, is essential in interpreting data obtained from plasma concentration monitoring. PMID- 3052010 TI - Rupture of the left ventricular free wall during acute myocardial infarction: analysis of 138 necropsy patients and comparison with 50 necropsy patients with acute myocardial infarction without rupture. AB - Clinical and necropsy findings in 138 patients (69 men and 69 women) with rupture of the left ventricular (LV) free wall during acute myocardial infarction (AMI) (rupture group) were compared with 50 patients who died during their first AMI without rupture (nonrupture group). The frequency of systemic hypertension (55 vs 52%), angina pectoris (13 vs 22%) and congestive heart failure (0 vs 0%) before the fatal AMI was similar for both rupture and nonrupture groups. Mean heart weights for men (479 vs 526 g) and women (399 vs 432 g) with and without rupture also were insignificantly different. LV scar before the infarct that ruptured was present in 18 patients (13%); previous necropsy studies of fatal AMI without rupture have indicated that 50% have LV scars. The rupture group had a significantly more frequent (p less than 0.01) lateral wall location of the infarct (12 vs 2%). The number of 3 major (right, left anterior descending and left circumflex) epicardial coronary arteries narrowed at some point greater than 75% in cross-sectional area by atherosclerotic plaque was significantly lower (p less than 0.01) in the rupture group (39 vs 58%). The percent of these 3 arteries totally occluded or nearly so (greater than 95% in cross-sectional area) by plaque also was significantly less (p less than 0.001) in the rupture group (24 of 198 arteries [12%] vs 38 of 144 arteries [26%]). Analysis of each 5-mm long segment of these arteries in each group disclosed that the rupture group had significantly less narrowing than the nonrupture group. Of the 3,287 five-mm segments of artery examined in the rupture group (66 patients), 512 (15%) were narrowed greater than 75% in cross-sectional area by plaque; in contrast, of the 1,848 five-mm segments in the nonrupture group (38 patients), 508 (28%) were narrowed to this degree by plaque (p less than 0.0001). Thus, rupture of the LV free wall primarily is a complication of the first AMI and is associated with considerably less amounts of coronary narrowing than fatal AMI without rupture. PMID- 3052011 TI - Retransplantation for severe accelerated coronary artery disease in heart transplant recipients. AB - Development of accelerated coronary artery disease (CAD) in the cardiac allograft is one of the major causes of late graft failure in heart transplant recipients. At the Stanford University Medical Center 356 heart transplant procedures were performed in 329 patients by the end of January 1985. Eighty-nine of these patients developed evidence of transplant CAD. Twenty retransplant procedures, including 2 third transplants, were performed in 19 of the 89 patients because of transplant CAD. The graft survival rates after the second transplant were 55%, 25% and 10% after 1, 2 and 5 years, respectively. Nine of these retransplant patients currently survive, the longest for 5.5 years. To examine potential risk factors for development of severe transplant CAD, these 20 retransplant procedures were compared with 113 transplant recipients who had no evidence of transplant CAD on annual coronary arteriograms. An excess of rejection episodes (3 +/- 2 vs 2 +/- 1 episodes/patient, p = 0.02), elevated total cholesterol (266 +/- 78 vs 225 +/- 47 mg/dl, p = 0.002) and higher low-density lipoprotein levels (176 +/- 88 vs 137 +/- 46 mg/dl, p = 0.009) were noted in the transplant CAD retransplant group. Five of 11 retransplant recipients who survived greater than 1 year again developed transplant CAD. Characteristic morphologic features and rapid progression of CAD in the second graft were similar to those in the primary graft.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3052012 TI - Phasic coronary artery flow velocity determined by Doppler flowmeter catheter in aortic stenosis and aortic regurgitation. AB - Phasic coronary artery flow velocity was recorded in 14 patients with aortic regurgitation (AR), 4 with aortic stenosis, 61 with other heart diseases and in 2 normal subjects by means of a bidirectional Doppler flowmeter catheter. The normal pattern of the phasic coronary artery flow velocity was characterized by a small forward flow during systole (S wave) and a large forward flow during diastole (D wave). The phasic coronary artery flow velocity in patients with AR showed increased S wave and decreased D wave. The area under the S-wave curve divided by the area under the D-wave curve (S/D ratio) in patients with AR increased (left coronary artery flow velocity 0.66 +/- 0.39, p less than 0.05; right coronary flow velocity 0.79 +/- 0.36, p less than 0.01) as compared with the S/D ratio in patients with other heart diseases (left coronary flow velocity 0.32 +/- 0.12; right coronary artery flow velocity 0.38 +/- 0.17). There was a tendency toward a relative positive correlation between S/D ratio values and AR cineangiographic grades. Decreased S/D ratios were observed in 4 patients with aortic stenosis. It is believed that no reports exist on phasic coronary flow velocity recorded in conscious patients who had aortic valve disease. PMID- 3052013 TI - Samuel A. Levine and his World War I experience. PMID- 3052014 TI - Richard D. Rowe, MD (1923-1988). PMID- 3052015 TI - The creatine kinase ratio: a useful means of detecting early peaking of the creatine kinase curve after acute myocardial infarction. PMID- 3052016 TI - Evaluation of the QRS complex on the standard 12-lead electrocardiogram in normal subjects 70 to 79 years of age. PMID- 3052017 TI - Specific skin lesions as the presenting symptom of hairy cell leukemia. AB - A 68-year-old Japanese man with a chief complaint of eczema-like dermatosis was diagnosed as having B-cell hairy cell leukemia (HCL) by demonstration of hairy cells in the skin lesions as well as in blood and bone marrow. He was treated with alpha-interferon, resulting in disappearance of skin lesions and reduction of his massive splenomegaly from 18 to 5 cm in about 14 months. Although specific skin lesions in HCL, shown by a review of the literature to occur in about 8% of cases, are not as uncommon as generally assumed, it is rare for HCL to present with specific skin lesions, the present case being only the second of its type mentioned in the literature. PMID- 3052018 TI - Acute myelogenous leukemia with eosinophilic differentiation and trisomy-1. AB - A 50-year-old woman presented with anemia and eosinophilia. Her bone marrow biopsy, peripheral blood, and clinical features were consistent with a diagnosis of an evolving acute myelogenous leukemia. Striking dysplastic eosinophilic differentiation associated with trisomy-1 was evident, and eosinophil granule major basic protein was detected in involved tissue. Trisomy-1 has not been previously reported in association with acute myelogenous leukemia showing eosinophilic differentiation. Intensive cytotoxic chemotherapy produced a short lived clinical and cytogenetic remission. At autopsy multiple tumor nodules composed of dysplastic eosinophil precursors and myeloblasts were evident in multiple organs. PMID- 3052019 TI - Malignant ameloblastoma with pulmonary metastasis and hypercalcemia. Report of an autopsy case and review of the literature. AB - A case of malignant ameloblastoma with hypercalcemia in a 67-year-old Japanese woman is presented. The tumor of the maxilla was removed and diagnosed as a follicular ameloblastoma. The tumor recurred in the lower orbita-zygoma region, and multiple tumors of the lungs and hypercalcemia were detected eight months after the second operation. The recurrent tumor resembled the primary tumor but was less well differentiated. Autopsy revealed widespread lung metastasis of the malignant ameloblastoma, nephrocalcinosis, and sigmoid colon cancer. Histologic examination showed the metastatic ameloblastoma to be composed of nests and strands of basaloid and spindle-shaped cells surrounded by columnar cells arranged in palisade formation, with focal areas of squamous differentiation and occasional cystic degeneration. Only two cases of malignant ameloblastoma with hypercalcemia have previously been reported. This is the first case of malignant ameloblastoma with hypercalcemia and sigmoid colon cancer. In addition, prostaglandin E2 assay revealed that ameloblastoma produces prostaglandin E2, which results in hypercalcemia. PMID- 3052020 TI - Follicular lymphoma mimicking progressive transformation of germinal centers: immunologic analysis of a case. PMID- 3052021 TI - Premature article--follicular lymphoma. PMID- 3052022 TI - Avoiding the problem of melanin pigment. PMID- 3052023 TI - Paraffin section immunophenotyping of non-Hodgkin's lymphoma, using a panel of monoclonal antibodies. AB - The monoclonal antibodies F8-11-13, 4KB5, MB1, and MB2 recognize largely B-cell restricted antigenic determinants that resist routine processing. Similarly, MT1, MT2, and UCHL1 react with fixation-resistant T-cell-restricted antigens. In order to evaluate the diagnostic potential of these antibodies, the authors have assessed their immunoreactivity with a series of 81 formalin-fixed and paraffin embedded non-Hodgkin's lymphomas (48 B-cell, 33 T-cell) encompassing a wide variety of histologic subtypes, which had been fully characterized by frozen section immunophenotyping. Ninety-six percent of B-cell lymphomas reacted with one or more of the B-cell-associated antibodies, whereas 100% of T-cell lymphomas reacted either with MT1, UCHL1, or both antibodies. MT2 was of no value in distinguishing between B- and T-cell lymphomas. None of the antibodies was entirely lineage specific; furthermore, a proportion of cases failed to react with one or more of the B- or T-cell-associated antibodies. Although these antibodies provide useful information in distinguishing between T- and B-cell lymphomas, the authors suggest that a panel of these antibodies is necessary for accurate determination of the histogenesis of these tumors. As with any immunohistochemical marker, interpretation of the immunostaining must be in the context of the morphologic features. PMID- 3052024 TI - Extranodal non-T-cell lymphoblastic lymphoma in adults. A report of two cases. AB - Typical adult cases of malignant lymphoma, diffuse, lymphoblastic type (MLLB), are of the T-cell immunophenotype and occur as mediastinal masses. The authors report two unusual adult cases with initial extranodal sites of involvement and non-T-cell immunophenotype that were originally misclassified. One patient presented with a right submaxillary salivary gland mass and the other presented with a T7-8 spinal cord compression resulting from a vertebral body tumor. A lymphoblastic leukemic phase developed in both patients after 15 months and 5 months, respectively. The bone marrow blasts of both patients had positive results for cluster designation (CD) 10, CD19, HLA-DR, and terminal deoxynucleotidyl transferase. Because both extranodal sites of presentation and non-T-cell immunophenotype are unusual in adult MLLB, these features may have contributed to the original misclassification of these lesions. PMID- 3052025 TI - Expansion of the lower arch concurrent with rapid maxillary expansion. AB - The effect of rapid maxillary expansion on the mandibular intercanine and intermolar widths during treatment and its stability after retention was studied. In addition, the relationships between the interarch change and the facial types and ages of the subjects of the sample were evaluated. The sample consisted of 17 cases for the study of intercanine width, and 22 nonextraction cases for the study of the intermolar width. Initial, final, at least 2-years postretention models, and initial lateral cephalograms were analyzed. Treatment and postretention changes for the intercanine width and the mesial and distal intermolar widths were calculated and tested for significance. Mean expansions of the intercanine width of 1.1 mm and of the intermolar width of 2.8 mm postretention were found to be statistically significant. There were no correlations found between the amount of increase in arch width and the facial types and ages of the subjects. PMID- 3052027 TI - Unexpected TMJ responses to functional jaw orthopedic therapy. AB - The activator, the Bionator, the Frankel, and, more recently, the Herbst appliances have enjoyed increasing popularity. Although an increase in mandibular growth has not been shown to be clinically consistent or always significant, the popularity of these appliances continues. Another clinical goal of these functional jaw orthopedic (FJO) appliances is to correct, maintain, or protect the integrity of the TMJ--specifically, to prevent posterior condylar displacement and/or anterior disk displacement. Because all FJO appliances anteriorly reposition the condyle during treatment, it is hoped that internal derangement problems may be resolved during treatment. Even though a single TMJ radiograph is not diagnostic in itself, multiple radiographs are helpful in monitoring the net changes in the condylar position during treatment. The relative (or net) change in the condylar position may provide clues to what occurred during treatment. However, TMJ responses are not always predictable during FJO treatment. Three case histories are presented that illustrate unexpected TMJ responses in which the condyles were still posteriorly displaced in spite of FJO treatment. Only 2% to 3% of the author's practice responds in this manner and no physiologic mechanism is suggested. These findings point out the complexity of the TMJ and its treatment, regardless of the appliance, and emphasize that no one approach to TMJ treatment will always be efficacious. PMID- 3052026 TI - Effects of appliance size, arch wire diameter, and alloy composition on the in vitro force delivery of the quad-helix appliance. AB - The quad-helix appliance is a fixed lingual arch wire appliance that produces slow maxillary expansion for the treatment of maxillary constriction or crossbite in the primary and mixed dentitions. Despite considerable success in the clinical treatment of patients, there is little quantitative information about the force delivery of this appliance. The purpose of this investigation was to characterize the in vitro force delivery of the quad-helix appliance. Appliances of four sizes were fabricated from Elgiloy blue and stainless steel wires with diameters of 0.032, 0.036, and 0.038 inch. The force delivery in grams for 8-mm activation was determined from the force-extension characteristics using an Instron mechanical testing machine. The results show that the force delivery of the quad-helix appliance is affected significantly by changes in the appliance size and arch wire diameter, but is independent of alloy type (stainless steel or Elgiloy blue). The quantitative results from this investigation should provide worthwhile information for clinical appliance selection and treatment of patients with varying arch dimensions. PMID- 3052028 TI - Group A streptococcal carrier state. PMID- 3052029 TI - Thyroid scanning, ultrasound, and serum thyroglobulin in determining the origin of congenital hypothyroidism. PMID- 3052030 TI - Abandonment, infanticide, and filicide. An overview of inhumanity to children. PMID- 3052031 TI - Plasma prorenin and renin in childhood and adolescence. AB - We studied 108 subjects (age range, 4 to 76 years) to determine the effect of age on prorenin (inactive renin), active renin, and plasma renin activity in normal children, adolescents, and adults. Children and adolescents had lower prorenin concentrations and higher plasma renin activity and active renin concentrations than did adults. Prorenin concentrations were positively correlated with age over the range of 4 to 76 years, while plasma renin activity and active renin concentration were negatively correlated with age. Plasma prorenin and active renin concentrations from umbilical cord blood samples obtained from 11 newborns and arterial samples obtained from five infants were higher than those in samples obtained from children or adults. PMID- 3052032 TI - Occult bacteremia in children with simple febrile seizures. AB - The controversy surrounding the diagnostic workup for simple febrile seizures has centered around the lumbar puncture. This focus has obscured the potential importance of other tests. A retrospective study was performed to determine the frequency of occult bacteremia in simple febrile seizures. In a pediatric emergency department, we identified 115 cases of simple febrile seizures in children treated as outpatients. Blood cultures were performed in 93 (81%) of 115 patients; five (5.4%) were positive. Children were less likely to have blood cultures performed if they were older than 24 months or had a medical history of simple febrile seizures. However, neither age nor history of febrile seizures affected the risk of bacteremia. These data suggest that patients with simple febrile seizures are at approximately the same risk for bacteremia as children with fever alone. Patients with simple febrile seizures should be treated in the same manner as other patients of the same age with regard to the performance of blood cultures. PMID- 3052033 TI - Disorders of higher cerebral function in preschool children. 1. AB - In preschoolers, disorders of higher cerebral function are most likely to present as inadequate development of language, while in school-age children, learning disabilities and attention deficit predominate. The main considerations in the differential diagnosis in preschoolers with inadequate language are hearing loss, mental deficiency, dysphasia, and autistic spectrum disorders. Attention to the child's ability to engage in symbolic play and communicate meaningfully is the key to this often baffling differential diagnosis. With the exception of definitive assessment of hearing, classic medical investigations are seldom informative because structural brain lesions and metabolic errors are much rarer etiologies than prenatal and genetic influences on brain development. While many children improve with age, the underlying deficit(s) usually persists. The role of the physician is to detect the developmental problem, give the parents a correct diagnosis, refer the child for appropriate investigation and intervention, and provide follow-up and counseling. Early diagnosis is essential for effective remediation. PMID- 3052034 TI - Diagnosis of Sjogren's syndrome in children. AB - We treated four children with clinical symptoms and laboratory findings suggestive of Sjogren's syndrome (SS). We also review the findings in 23 children with the diagnosis of SS whose cases were reported in the literature. We propose that the following criteria for the diagnosis of SS, which are mostly used in adults, should also be applied to children: (1) keratoconjunctivitis evidenced by a Schirmer test and a quantitative rose bengal test; (2) xerostomia shown by a decreased basal and stimulated salivary flow; (3) lymphocytic infiltration in a minor salivary gland biopsy specimen with at least two foci per 4 mm2; (4) laboratory evidence of a systemic autoimmune disorder on the basis of a rheumatoid factor of 1/160 or greater, antinuclear antibody of 1/160 or greater, or extractable nuclear antigen antibodies. Only close observation and long-term follow-up of these patients will allow a better insight in the natural history of SS in children. Those children who do not fulfill these diagnostic criteria also need close and prolonged follow-up study: one of the possibilities is that their conditions will ultimately evolve toward definite SS. PMID- 3052035 TI - Picture of the month. Epidermolytic hyperkeratosis. PMID- 3052037 TI - The pursuit of intoxication: our 100 century-old romance with psychoactive substances. PMID- 3052036 TI - Psychopathology in substance abusers: diagnosis and treatment. PMID- 3052038 TI - Adolescent alcohol and drug abuse and its consequences--an overview. PMID- 3052039 TI - Substance abuse among general psychiatric patients: place of presentation, diagnosis, and treatment. AB - This paper reviews the literature on patients presenting for general psychiatric treatment who are also substance abusers. Place of presentation, diagnosis, and treatment are considered. A considerable portion of patients seen in emergency rooms, as much as half in some settings, are substance abusers, and over a third of general psychiatry admissions have been found to have their presenting problems materially influenced or precipitated by substance abuse. Substance abuse is also frequently found among psychiatric inpatients. Diagnostically, the differentiation of general psychiatric and addictive syndromes can be difficult: primary and secondary affective disorder from consequences of long-term substance abuse; and self-medication patterns from primary general psychiatric syndromes. Treatment studies are often focused on concomitant psychotherapeutic management for patient being treated for addiction. Often, emphasis is placed on pharmacotherapy for enhancing outcome in the dually diagnosed. Qualitatively, new options tailored to this population still remain to be studied, however, as do the changes necessary in the treatment system to assure proper long-term management. PMID- 3052040 TI - Sudden death and anomalous origin of the coronary arteries from the aorta. A case report and review. AB - Based on a case report of sudden death in a young boy, this paper reviews the available information concerning the various combinations of anomalous origins of coronary arteries and associated sudden death. Left coronary arteries arising from the right sinus of Valsalva and passing between the aorta and pulmonary arteries are often associated with sudden death and myocardial ischemia in young people. Although right coronary arteries originating from the left sinus of Valsalva and passing between the aorta and pulmonary artery are less frequently associated with symptoms, this condition may be associated with sudden death. The incidence of symptoms associated with other anomalous origins is also discussed. PMID- 3052041 TI - Birchall and Benwell. Murder in a Canadian swamp. AB - On 21 February 1890, two woodsmen working in densely wooded Blenheim Swamp in southern Ontario, Canada, stumbled upon the dead body of a young Englishman who had been killed by two gunshot wounds to the head. He was identified as Frederick Benwell by a married couple, the Birchalls, who had traveled with him and another man, Pelly, from England to New York City by ship about 1 week before. The Birchalls lied regarding Benwell's subsequent movements. Questioning Pelly revealed to Detective John Murray that both Pelly and Benwell had replied separately to Birchall's advertisement for young men of means to become partners with him in a large Canadian farm, a deposit first being required. Birchall had been observed taking Benwell from Buffalo, New York, to Blenheim Swamp, for in reality there was no farm. There he had shot Benwell, leaving the body partially exposed. He then tried unsuccessfully to lure Pelly to his death in Niagara Falls. The Birchalls were arrested. Reginald Birchall was tried, convicted, and hanged. His attempt, including murder, to turn the so-called farm pupil colonization scheme to his own benefit had been frustrated by the dogged work of the master Canadian detective Murray. PMID- 3052042 TI - Sudden death due to ruptured pancreaticoduodenal artery aneurysm. AB - Superior pancreaticoduodenal artery aneurysms are rare; their rupture often leads to sudden death. We report a case of a 59-year-old hypertensive man who died of a massive retroperitoneal hemorrhage following rupture of an aneurysm of the superior pancreaticoduodenal artery. We also discuss this unusual vascular lesion and review the pertinent literature. PMID- 3052043 TI - Phenotypic heterogeneity of erythroblasts in erythroblastic leukemia revealed by monoclonal antibodies. AB - Eight cases each of erythroleukemia (AML-M6) and erythroblastic crisis of chronic granulocytic leukemia (CGLBC-E) were immunophenotyped with the help of a panel of lineage-associated monoclonal antibodies (McAbs). The latter included those reactive with erythroid progenitor (BFU-E and CFU-E) and erythroid precursors at different stages of maturation. In six of eight cases of AML-M6, erythroblasts revealed an immature phenotype, as evident from reactivity of the blast cells with McAbs directed against the earlier stages of erythroid maturation. One case had the phenotype of CFU-E, and in the remaining case of AML-M6 the erythroblasts showed a "mature" surface antigenic profile. This immunophenotypic spectrum was unrelated to the morphologic maturity of the erythroblasts. In two cases of CGLBC E, an early erythroblastic phenotype was observed, while in as many cases a "mature" phenotype was present. Four of eight cases, however, revealed a mixed, erythroid plus myeloid phenotype. In one of the four cases, two separate blast populations, which represented erythroblasts and myeloblasts, could be identified. In the remaining three cases the blasts were morphologically homogeneous and undifferentiated. High incidence of HLA-DR positivity in the latter three cases suggests the primitive nature of blasts cells and their closeness to the putative "bipotent" myeloid stem cell. Our study has shown phenotypic heterogeneity of blast cells in AML-M6 and CGLBC-E. PMID- 3052044 TI - Multiple myeloma: relationship between light chain isotype suppression, labelling index of plasma cells, and CD38 expression on peripheral blood lymphocytes. AB - In this study we have investigated the relationship between the labelling index of plasma cells, the expression of CD38 positive lymphocytes in the peripheral blood, and light chain isotype suppression. This study confirms the relationship between plateau-phase disease and light chain isotype suppression (LCIS) and documents an inverse relationship between LCIS and CD38 positive lymphocytes (.001 less than P less than .01), which is similar to the relationship we have described with the expression of CD10 positive lymphocytes. PCA-1 is rarely expressed in the peripheral blood of patients with myeloma and does not fulfill a role as a marker of active vs. stable disease. There is no relationship between the labelling index of plasma cells and LCIS, because many patients can enter a stage of progressive disease and yet have a labelling index of less than 1% at that time, although a labelling index less than 1% is present in the majority of patients with LCIS. beta-2-microglobulin (beta 2M) also fails to differentiate these two phases of disease in myeloma and does not have a relationship with LCIS, CD38 expression, or CD10 expression. These data suggest that myeloma, like chronic granulocytic leukemia (CGL), can be considered as having two phases of disease: a stable or chronic phase disease, as identified by the presence of LCIS, the absence of CD10 and CD38 positive lymphocytes in the peripheral blood, and a low labelling index, and progressive disease, which is associated with the loss of LCIS and of, CD10 and CD38 positive lymphocytes in the peripheral blood and a high labelling index, although in many cases of progressive disease, the labelling index may also be low. beta 2M does not differentiate between these states. PMID- 3052045 TI - Recurrent spontaneous bacterial peritonitis in a patient with polycythemia vera. AB - Spontaneous bacterial peritonitis (SBP) is an infectious process that usually occurs in patients with cirrhosis. There are few reports of SBP in patients with other pathologies such as nephrotic syndrome, acute and chronic hepatitis, cardiac ascites, and ascites secondary to neoplastic disease. We report a patient with polycythemia vera in whom recurrent episodes of SBP occurred 8 months following a portacaval shunt operation for Budd-Chiari syndrome. Conceivably, the polycythemia vera (PV) complicated by hepatic vein thrombosis and portacaval shunt resulted in significant loss of hepatic reticuloendothelial system function and predisposed the patient to bacterial peritonitis. PMID- 3052046 TI - Results of coronary artery bypass grafting in end-stage renal disease. AB - We examined the results of coronary artery bypass grafting (CABG) in patients with end-stage renal disease and symptomatic ischemic heart disease who had significant arteriosclerotic narrowing of one or more coronary vessels between 1970 and 1984. Twenty-four such patients underwent bypass grafting, 20 dialysis patients and four who had been transplanted. Bypass grafting completely or partially relieved symptoms in 83%. The hospital mortality associated with this surgery for the 20 dialysis patients was 20% compared with a lower overall hospital mortality for bypass grafting in nondialysis patients of 1.3%. Greater hospital mortality was noted for patients over age 60 undergoing bypass grafting, 33.3% v 1.9% in nondialysis patients. In this study, the most significant factor associated with mortality was older age. We conclude that bypass grafting has an acceptable mortality in younger end-stage renal disease patients anticipating or having had renal transplantation, but it is associated with a high hospital mortality in older dialysis patients. PMID- 3052047 TI - Characterization of interstitial infiltrating cells in Berger's disease. AB - The role of infiltrating blood-borne cells in the pathogenesis of renal damage in human glomerulonephritis is under active investigation. We have evaluated leukocyte infiltrates (number of cells/mm2) in the renal interstitium of 21 patients with Berger's disease and eight normal kidneys with monoclonal antibodies and a four-layer immunoperoxidase technique. In our population of patients, the number of infiltrating T-lymphocytes (OKT11+ cells) was significantly higher (median, 132) than in the normal kidneys (median, 60). This increase was mainly due to T-suppressor/cytotoxic lymphocytes (OKT8+ cells; median, 68), while T-helper/inducer lymphocytes (Leu 3A+ cells) and monocytes were in the normal range. T-lymphocyte infiltration was more marked in ten patients with impaired glomerular filtration rate (GFR) at the time of biopsy (median, 167) than in patients with normal GFR (median, 88). In addition, ten patients who showed deterioration of renal function during the subsequent follow up, whatever their serum creatinine levels at the time of biopsy, had significantly more total T cells (median, 269), OKT8+ cells (median, 143), and Leu 3A+ cells (median, 105) than 11 patients with persistently stable GFR and normal controls. More data are necessary to establish whether this T-lymphocyte infiltration is the consequence of a cell-mediated mechanism acting in the interstitium, concomitant with the immune-complex-mediated mechanism acting in the glomerulus, or is a nonspecific consequence of the tubulointerstitial damage induced by the immunologically mediated glomerular disease. PMID- 3052048 TI - B2-microglobulin and associated amyloidosis presenting as bilateral popliteal tumors. AB - Chronic hemodialysis has led to the prolongation of life in many patients with end-stage renal disease, but has also allowed for the development of new diseases that are a consequence of this clinical setting. B2-microglobulin accumulation leading to systemic amyloidosis may be the most recent disease in this category. This case report documents the development of bilateral popliteal tumors in a patient undergoing chronic hemodialysis for 9 years. Removal of one of the tumors and pathological examination demonstrated amyloid that was positive for B2 microglobulin by specific antibody testing. This case adds further support to the suggestion that B2-microglobulin amyloidosis in chronic hemodialysis patients is truly a systemic disorder. The development of popliteal tumors, particularly in proximity to joints, in a chronic hemodialysis patient, must include amyloidosis in the differential diagnosis. PMID- 3052049 TI - Review and hypotheses: somatic mosaicism: observations related to clinical genetics. PMID- 3052050 TI - Effect of various therapeutic approaches on plasma potassium and major regulating factors in terminal renal failure. AB - PURPOSE: The development of life-threatening hyperkalemia poses a risk for patients with chronic preterminal renal failure. Various therapeutic options have been suggested for hyperkalemic emergencies in these patients; to date, however, no study has evaluated the relative efficacies of these measures in the presence of renal failure. Our goal was to examine the acute effects of a variety of therapeutic approaches, as well as those of hemodialysis, on plasma potassium levels in a hemodialysis population. PATIENTS AND METHODS: Ten patients with terminal renal failure undergoing maintenance hemodialysis were enrolled in the study. Blood gas parameters and plasma sodium, potassium, glucose, osmolality, renin, aldosterone, epinephrine, norepinephrine, dopamine, and insulin were measured before, during, and after 60-minute infusions of bicarbonate, epinephrine, and insulin in glucose, and before, during, and after performance of regular hemodialysis for one hour. RESULTS: Hypertonic as well as isotonic intravenous bicarbonate (2 to 4 mmol/minute) induced a marked rise in plasma bicarbonate and pH, but failed to lower the plasma potassium level (5.66 versus 5.83 mmol/liter before and after). Epinephrine, 0.05 microgram/kg/minute administered intravenously, decreased plasma potassium only slightly from 5.57 to 5.25 mmol/liter, and five patients showed no decline. On the other hand, insulin in glucose, 5 mU/kg/minute intravenously, effectively lowered plasma potassium levels from 5.62 to 4.70 mmol/liter, and hemodialysis induced the most rapid decline from 5.63 to 4.29 mmol/liter. Plasma aldosterone was elevated before treatment; it correlated with plasma potassium and dropped during intravenous bicarbonate administration or hemodialysis. Pretreatment plasma renin activity, insulin, epinephrine, norepinephrine, and dopamine levels were generally normal. CONCLUSION: We conclude that in patients with terminal renal failure undergoing maintenance hemodialysis, intravenous bicarbonate is ineffective in lowering plasma potassium rapidly, and epinephrine is effective in only half the patients, whereas insulin in glucose is a fast and reliable form of therapy for hyperkalemic emergencies. Plasma aldosterone levels are appropriate in relationship to plasma potassium levels, and levels of other potassium influencing hormones are generally normal. PMID- 3052051 TI - A randomized, double-blind, placebo-controlled study of verapamil and metoprolol in treatment of multifocal atrial tachycardia. AB - PURPOSE: Multifocal atrial tachycardia is a difficult arrhythmia to treat. Patients not showing a response to the correction of predisposing conditions present a therapeutic dilemma. To assess the efficacy of two agents reported to be effective in this condition, verapamil, metoprolol, or placebo was given intravenously in a randomized, double-blind trial. PATIENTS AND METHODS: Thirteen patients meeting inclusionary criteria were enrolled. Therapeutic response was defined as conversion to sinus rhythm, a decline in the ventricular rate of 15 percent or more [corrected], or a ventricular rate of less than 100 beats/minute. Four male and nine female patients having a mean age (+/- SD) of 81.9 +/- 14.2 years were enrolled. Automated serum chemistries, complete blood cell count with differential, arterial blood gas values, and serum digoxin and theophylline levels were determined and a 12-lead electrocardiogram was obtained at the start of the trial. Following the completion of each phase of the study, a repeat physical examination was performed, and arterial blood gas values and an electrocardiogram were obtained. The trial was designed to run for two days. RESULTS: Two of 10 (20 percent), four of nine (44 percent), and eight of nine (89 percent) showed a response to placebo, verapamil, or metoprolol, respectively. Mean slowing of ventricular rate was 3.4, 7.3, and 24.5 percent for placebo, verapamil, and metoprolol, respectively (p less than 0.01 for metoprolol versus placebo). Five patients who showed a response to metoprolol had failed to have a response to verapamil. CONCLUSION: We conclude that metoprolol appears to be more effective than verapamil in treating multifocal atrial tachycardia. However, careful patient selection is necessary in its use. PMID- 3052052 TI - New perspectives on the pathogenesis of rheumatoid arthritis. AB - In the pathogenesis of rheumatoid arthritis, locally produced antibodies complex with an inciting antigen, yet to be identified, within the joint and activate the complement system, resulting in articular inflammation mediated primarily by polymorphonuclear leukocytes and their products. Chronic inflammatory cells then produce soluble factors that induce both tissue destruction and inflammation. A major issue is how and why apparently normal immune responses in the acute stage progress to chronic inflammation in subsequent months to years. Although it is often assumed that the initial etiologic agent, persisting in the joint or at an extra-articular site, is responsible for continued synovitis, this need not be the case. It is possible that once the inciting agent is cleared from the joint through a normal immune response, the presence of activated cells rich in surface class II histocompatibility (Ia) antigens could, under the influence of multiple genetic or environmental factors, become the target of autoimmune attack. Alternatively, the process might result from the interactions of synovial lining cells and their products with T cells assuming a secondary role. Further research into the relative contributions of soluble products, T helper and suppressor subsets, synoviocytes, and antigen determine which model is correct. PMID- 3052053 TI - Hydroxychloroquine treatment of rheumatoid arthritis. AB - A variety of placebo-controlled and open studies have demonstrated the effectiveness of hydroxychloroquine in the treatment of rheumatoid arthritis. The excellent responses to recurrent treatment in a sample patient illustrate the value of hydroxychloroquine. Because low daily doses of hydroxychloroquine are associated with greater ophthalmologic safety, it would be advantageous to use the smallest effective daily dose, but there are no published controlled efficacy studies using daily doses of less than 400 mg. Hydroxychloroquine may best be employed to treat patients with new onset of disease or those in whom disease is not rapidly progressive. Great potential exists for the use of hydroxychloroquine in combination therapy, but optimal utilization of combination regimens will require performances of additional controlled studies. PMID- 3052054 TI - Newer laboratory parameters for the diagnosis of rheumatic disease. AB - Measurement of serum autoantibodies is a useful adjunct in the diagnosis of connective tissue diseases. Approximately 15 percent of patients with rheumatoid arthritis are seronegative for rheumatoid factor by standard assays. The development of enzyme-linked immunosorbent assays for additional isotypic and idiotypic determinants may expand the diagnostic sensitivity of rheumatoid factor measurements. Although extremely sensitive, the finding of antinuclear antibodies is not highly specific for systemic lupus erythematosus and related disorders; the ability to rapidly screen serum against newly characterized specific nuclear antigens has been helpful in the diagnosis of newly described systemic lupus erythematosus subtypes, overlap syndromes, Sjogren's syndrome, and scleroderma variants. Women with unexplained recurrent fetal loss and patients with unexplained thrombotic episodes should be screened for the presence of the anticardiolipin antibody. Glycoproteins (C-reactive protein, alpha-1 glycoprotein, fibronectin) may prove useful as indicators of rheumatic disease activity and the efficacy of therapeutic intervention. PMID- 3052056 TI - Antimalarial agents compared with or in combination with other disease-modifying antirheumatic drugs. AB - Available published comparisons of antimalarial drugs with other disease modifying antirheumatic drugs (DMARDs) are reviewed, as well as reports of combinations of DMARDs, in the treatment of rheumatoid arthritis and juvenile rheumatoid arthritis. These reports suggest that antimalarial drug toxicity is less than that of parenteral gold, D-penicillamine, auranofin, or dapsone. Its efficacy appears to be equal to or slightly less than that of most comparison DMARDs. Combinations of DMARDs with antimalarials are probably used by a substantial proportion of rheumatologists, but the few prospective, double-blind studies with a balanced design show little or no advantage to combination DMARD therapy. Open, nonrandomized studies and preliminary reports suggest that combination therapy may be useful, usually when another DMARD is added to one or more DMARDs to which the patients did not have a satisfactory response. A few large, prospective, double-blind, randomized studies comparing hydroxychloroquine with several other DMARDs and placebo are needed to establish more confidence in the relative efficacy and utility of hydroxychloroquine. Although the rationale for combination DMARD therapy is attractive, additional preliminary studies of the various possible combinations and doses are needed before definitive trials of promising combinations can be justified. PMID- 3052055 TI - Role of disease-modifying antirheumatic drugs versus cytotoxic agents in the therapy of rheumatoid arthritis. AB - Disease-modifying antirheumatic drugs are used to modify or alter the rheumatoid arthritis disease process. Disease-modifying antirheumatic drugs do not demonstrate the characteristic analgesic, antipyretic, and anti-inflammatory actions of the nonsteroidal anti-inflammatory drugs, since weeks or months of treatment are required before clinical benefit is observed. Although they have not been proved to delay, prevent, or reverse articular damage, therapy with disease-modifying antirheumatic drugs, when successful, is associated with decreased pain and joint swelling and improved function. Disease-modifying antirheumatic drugs and cytotoxic agents should not be considered as routine treatment for patients with rheumatoid arthritis. Before disease-modifying antirheumatic drug therapy is implemented, an optimal basic program of physical therapy, rest, and nonsteroidal anti-inflammatory drugs should be implemented, and it must be documented that the patient still has sufficient disease to justify the costs, risks, and benefits of these treatments. Drugs that are approved by the Food and Drug Administration (FDA) are preferred over nonapproved drugs. Hydroxychloroquine, parenteral gold salts, oral gold, D-penicillamine, and the cytotoxic drug azathioprine are the FDA-approved disease-modifying antirheumatic drugs for use in rheumatoid arthritis. Many, not all, rheumatologists would employ hydroxychloroquine as the first-choice disease modifying antirheumatic drug in patients who have early, mild, and nonerosive disease; treatment should be continued for six months before being abandoned for lack of efficacy, and appropriate ophthalmologic examination every four to six months is indicated. An alternative would be auranofin, whose efficacy approaches that of parenteral gold, yet may be safer. For patients who have severe active rheumatoid arthritis, especially with erosive changes, parenteral gold salts are usually a first choice. D-penicillamine is also effective in controlling the signs and symptoms of rheumatoid arthritis, but serious toxicity may occur. Azathioprine might be considered a competitor to D-penicillamine, although the FDA approval restricts its use to patients who have failed gold therapy. Two cytotoxic drugs that are not FDA approved are methotrexate and cyclophosphamide. Methotrexate can be very effective, but its side effects, particularly pulmonary and hepatic, must be carefully monitored. Cyclophosphamide is generally considered too toxic for use in patients with rheumatoid arthritis, although it may be helpful in patients with systemic rheumatoid vasculitis or patients who have failed all other therapies.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3052057 TI - Hydroxychloroquine and postoperative thromboembolism after total hip replacement. AB - Evidence for the usefulness of hydroxychloroquine as prophylaxis against thromboembolism after total hip replacement is examined. This agent causes reduction in red blood cell aggregation without prolonging the bleeding time in humans and, experimentally, reduces the size of the thrombus. There is a variably demonstrable reduction in platelet aggregation and blood viscosity in humans. After hip replacement, the venographic incidence of deep vein thrombosis is not influenced by hydroxychloroquine. The incidence of fatal pulmonary embolism appears to be reduced, but a controlled trial against placebo does not exist. There are no serious or irreversible side effects, and wound healing is not affected by the administration of this drug. Hydroxychloroquine remains the prophylaxis of choice in use at the Hip Center, Wrightington Hospital, England, 13 years after its introduction. PMID- 3052059 TI - Inhibition of renin by plasma linoleic acid. AB - The authors have demonstrated previously that human plasma contains an inhibitor(s) of the enzymatic activity of renin. The purpose of this study is to identify the circulating renin inhibitor. Plasma initially was fractionated with preparative sephacryl S-300 chromatography. A fraction with low protein content inhibited the in vitro enzymatic activity of human renin by 76%. The inhibitory activity of this fraction was not altered by boiling and/or acidification. This fraction was applied to an affinity column, using purified mouse submaxillary renin as ligand. The inhibitor was recovered after elution with hypertonic NaCl. Linoleic acid, a previously identified renin inhibitor, was present in this fraction. The authors conclude that circulating linoleic acid inhibits renin. Conceivably, the overall activity of the renin-angiotensin-aldosterone cascade may be modified by alterations of plasma fatty acid concentrations. PMID- 3052058 TI - The role of epidermal growth factor in skin diseases. PMID- 3052061 TI - A review of ceftriaxone: a long-acting cephalosporin. PMID- 3052060 TI - The premise, promise, and perils of the prevention of lethal ventricular tachyarrhythmias. AB - Sudden cardiac death caused by ventricular tachyarrhythmias claims about 360,000 lives a year in the United States. The premature ventricular complex (PVC) hypothesis has been the cornerstone for understanding this problem, but it is now recognized as an incomplete explanation for this catastrophy. The recognition of the importance of structural heart disease in this process has led to the development of the Substrate Hypothesis as an alternative explanation. In this construct, PVCs may trigger lethal arrhythmias but only if an abnormal electrophysiologic substrate is present. This hypothesis more completely describes the pathophysiology of the process, provides the basis for understanding the value and limitations of the techniques used for risk assessment and management, and helps clarify the potential endpoints and potential adverse effects of therapy to prevent arrhythmias. Since no single diagnostic technique is ideal and no therapeutic modality is universally effective, an approach to the management of this problem must be multidimensional and based on a firm understanding of the actual risk of a life threatening arrhythmia, the potential but unproven benefits and uncertain endpoints of drug therapy, the cost, and the potential for arrhythmia exacerbation or significant side effect. PMID- 3052062 TI - FG syndrome update 1988: note of 5 new patients and bibliography. AB - At the eve of its mapping, the pre-molecular picture of the FG syndrome is heavily biased towards the severe end of the phenotypic spectrum because present knowledge is largely based on propositi. It is an X-linked, incompletely recessive, complexly pleiotropic syndrome with considerably variable expressivity. Though a true multiple congenital anomalies/mental retardation (MCA/MR) syndrome, severe malformations are uncommon and involve mostly the anus (60%) and non-colonic GI defects (33%), hypospadias (25%), cleft palate (6%), rarely a congenital heart defect. The complex CNS dysfunctions of congenital hypotonia and all of its sequelae, MR, and occasional seizures, must be attributed to a developmental CNS defect which is rarely demonstrated at pre mortem, and which is known to involve agenesis of the corpus callosum in some 25% of appropriately studied patients (mostly propositi). Thus, the diagnosis is largely made on a specific constellation of minor anomalies and mild malformations in a hypotonic boy with severe constipation and a very characteristic facial appearance and behavioral phenotype. In about 1/3 of cases, carrier manifestations may be detected physically. New hemizygote manifestations seen in this review of 5 new patients include abnormal eruption of teeth, diastasis between upper central incisors, apparent gynecomastia, cleft lip, and nasolacrimal and helicine fistulae. Only a half hundred or so FG syndrome patients are known, but we suspect the syndrome is much more common than realized, and because of the unfortunate recurrence risk potential, deserves careful consideration in every appropriate case. RFLP mapping studies are urged in order to aid diagnosis of "mild" cases, and prenatal and carrier detection. PMID- 3052063 TI - Bibliography on X-linked mental retardation, the fragile X and related subjects IV (1988). PMID- 3052064 TI - A controlled trial of stimulant medication in children with the fragile X syndrome. AB - Attentional deficits and hyperactivity frequently are major problems for fra(X) boys. This study evaluated the effectiveness of 2 stimulant medications, methylphenidate and dextroamphetamine compared to placebo in 15 children (13 males, 2 females) with the fra(X) syndrome. A double-blind crossover design was used with outcome measures which included parent and teacher behavior checklists, a controlled observation period, continuous performance tasks and an actometer measure of movement. When the children were treated with methylphenidate only, improvement was seen in socialization skills and attention span according to teacher checklists. Ten children were clinically considered responders and treatment was continued after the study was completed. PMID- 3052065 TI - Folic acid treatment of fragile X males: a further study. AB - Investigations of the effect of high dose folic acid treatment of fragile X syndrome in males has produced mixed results. However, no study had examined the possible drug effects of folic acid on non-fragile X control males. Therefore, we examined the effect of folic acid on fragile X males using non-fragile X control males. Subjects were assigned randomly to an ABA or BAB design. Duration of either folic acid or placebo condition was 4 months. Folic acid or placebo was given in a double-blind fashion. At the end of each condition, the subjects' behavior was assessed. At the end of the study, parents were asked to complete a questionnaire. Using parents' responses, we examined 22 items on the Autistic Descriptors Checklist and two subscales from the Vineland Adaptive Behavior Scale which corresponded to areas of behavior parents' noted to have shown improvement. We did not find significant differences between fragile X males and control males, within subjects, nor across folic acid and placebo conditions. Thus, our follow-up study confirms and extends our original findings, as well as those of other researchers: namely, that no dramatic changes in behavior result from high dose folic acid. Moreover, subtle improvements observed in earlier investigations were not confirmed. PMID- 3052066 TI - Constitutive fragile sites in fra(X) individuals. AB - Recently, it was proposed that the constitutive fragile site at 3p14 be used as an "internal control" to indicate the effectiveness of the FUdR fragile site induction system. We have tested this hypothesis by determining the frequency of constitutive fragile sites at 1p31, 3p14, and 16q23 in cultures from 42 known fra(X) individuals. At least 50 cells were analyzed from each case. Seventy-four percent (31/42), 95% (40/42) and 90% (38/42) of the fra(X) individuals exhibited frequencies of less than 4% at constitutive fragile sites 3p14, 1p31 and 16q23, respectively. Of the 42 individuals tested, 12 or 28.6% showed no fragility at any of the 3 sites studied. On the other hand, at least one constitutive fragile site was observed in 50 cells studied from over 70% of the 42 people studied. It is suggested that "positive controls" continue to be used, while at the same time recording all fragile sites to identify a combination of constitutive fragile sites that may serve as an internal control indicator, and that DNA marker studies be used to complement cytogenetic testing. PMID- 3052067 TI - In situ nick translation of the fragile X region. AB - At the present time, the molecular nature of the fragile site at Xq27.3 is not well understood. To examine the sensitivity of this region to DNAase I, in situ nick translation was performed on metaphase chromosomes from a fragile X (fra(X] positive individual. In this technique DNAase I is used to nick regions of chromosomal DNA that are in "open" conformation. Biotinylated dUTP was incorporated by nick translation at these sites. The incorporation was identified by double antibody labeling and avidin-horseradish peroxidase staining. Spreads, which had been stained with this technique, were photographed and subsequently trypsin-Giemsa G banded (post-GTG banded) for chromosome identification. In 36 of 44 (82%) fra(X) positive male cells, the region distal to fra(X) (q27.3) was prominently stained in contrast to its light staining appearance in GTG preparations. The fragile site itself was outlined more clearly than can be achieved by GTG or homogeneous staining. When autosomal fragile sites were induced by the addition of 1.5 microM aphidicolin 17 hours prior to harvest, 24 of 27 (89%) fragile sites on the ends of autosomes were prominently stained in regions distal to the break. Because the fra(X) and autosomal fragile regions behaved similarly, this suggests that they have a similar conformation. Thus, while autosomal and Xq27.3 fragile sites are strongly induced by different means, the organization of these sites and the regions distal to them appear to be similar. PMID- 3052068 TI - Induction of the fra(X) in amniotic fluid cells by excess thymidine. PMID- 3052069 TI - Is there a fragile(X) negative Martin-Bell syndrome? AB - During the course of the preventative screening program for the fra(X) syndrome, we identified 32 men with the phenotype but who were fra(X) negative. These were reviewed and none fitted the full criteria, so we were unable to confirm the existence of the fra(X) negative Martin-Bell syndrome. The literature and 4 families previously thought to have the fra(X) negative Martin-Bell syndrome were also reviewed. We were unable to make a concrete diagnosis of the fra(X) negative Martin-Bell syndrome. PMID- 3052070 TI - Screening developmentally disabled male populations for fragile X: the effect of sample size. AB - The fra(X) or Martin-Bell syndrome is the most common cause of inherited mental retardation (MR) in males. It is also associated with a variety of unusual behavioral and developmental disorders. Recent studies found great variability in the estimated strength of association between "autism" and the fra(X) syndrome, but not between MR and fra(X). We examined 31 studies which investigated the association of fra(X) syndrome with either MR or "autism" and found that the conclusion of those researchers could be significantly affected by sample size. Different behavioral and cytogenetic protocols will also influence the strength of association between fra(X) and autism. PMID- 3052071 TI - Fragile X families in a northern Swedish county--a genealogical study demonstrating apparent paternal transmission from the 18th century. AB - Eleven families including 35 cases with fra(X) mental retardation (MR) were traced genealogically using the Research Archives at Umea University. Seven of the cases were women with fra(X). All of the families originated partly or totally from the county of Vasterbotten. It was possible to link 7 of the index families to common ancestors over an 8-11 generation span. The remaining 4 families were not traced to the same ancestors. However, they were linked together pair-wise over a 7-8 generation span. Transmission of the fra(X) mutation was studied in these families. In the pedigree analyses, priority was given to maternal transmission. In 2 families the fra(X) mutation was transmitted solely through females over 7 or 8 generations respectively. Within 9 families the mutation was transmitted by males in 2-5 generations in order to reach common ancestors. PMID- 3052073 TI - When chronic illness calls for more than chronic care. PMID- 3052072 TI - Dismantling the Medicare home health benefit. PMID- 3052074 TI - Heart transplant. Impact on CCU nurses. PMID- 3052075 TI - Fetal surveillance in insulin-dependent diabetic pregnancy: predictive value of the biophysical profile. AB - Ninety-eight insulin-dependent diabetic pregnancies underwent monitoring by means of 978 biophysical profiles from 28 weeks' gestation until parturition. Only 2.9% of the 978 tests had abnormal results (score less than or equal to 7). When performed within 2 days before birth, a normal biophysical profile predicted the 1-minute Apgar score to be normal in 92% and 5-minute Apgar score in 99%. When all biophysical profiles ever performed were included, the predictive value improved to 100%. The baby's first cry within 1 minute after birth was predicted in 95%. Furthermore, the predictive value of a normal biophysical profile regarding the absence of ominous intrapartum cardiotocographic patterns was excellent (95%). The specificity was in general good (80% to 90%), but the predictive value of abnormal test results and sensitivity were almost without exception poor. It seems that the very low rate of abnormal biophysical profiles indicates that obstetric interventions were made immediately after the occurrence of the first sign of fetal jeopardy; thus improved results were obtained. PMID- 3052076 TI - The use of transvaginal sonography for evaluation of postmenopausal ovarian size and morphology. AB - Ultrasonic evaluation has been suggested as a possible screening tool for early changes in ovarian morphology. This study uses transvaginal sonography to evaluate the ovaries in postmenopausal women who were scheduled for gynecologic surgery unrelated to adnexal disease. The findings of ultrasonic ovarian examination are compared with the findings at surgery and the pathologic evaluation of the ovaries. Nine (17.3%) abnormal ovaries were identified by ultrasonography and at surgery and were confirmed at pathologic examination. Among the abnormal ovaries there were one malignancy (10%) and two neoplasms with known malignant potential (20%). One ovary that was identified to have microscopic areas of Brenner tumor cells at pathologic examination was described as normal by both ultrasound and surgical evaluation. The sensitivity (90%) and specificity (100%) of vaginal sonography were the same as that of gross examination of the ovary at the time of surgery. We conclude that vaginal sonography is a reliable tool in the detection of early abnormalities in the postmenopausal ovary. PMID- 3052077 TI - Immunocytochemical HBsAg evidence in placentas of asymptomatic carrier mothers. AB - We report three chronic asymptomatic HBsAg carrier mothers. We studied their placentas via application of the immunohistochemical method to identify HBsAg by peroxidase-antiperoxidase techniques. All three placental specimens showed a positive HBsAg reaction. Due to the strong cytoplasmatic reactivity of the cytotypes, we deduced their active participation in the infection, proving that virosis occurs via transplacental circulation. PMID- 3052078 TI - The value of preoperative serum CA 125 levels in patients with a pelvic mass. AB - Serum CA 125 measurements were obtained before operation in 250 consecutive patients admitted with the diagnosis of a pelvic mass. We examined and operated on all patients. Elevated preoperative serum CA 125 levels were more predictive of malignancy in postmenopausal than in premenopausal women (0.86 vs 0.67), but normal levels did not preclude malignancy in either group. Possible limitations of routine measurement of serum CA 125 in the work-up of patients with a pelvic mass are discussed. PMID- 3052080 TI - Computed tomographic features of puerperal ovarian vein thrombosis. AB - Puerperal ovarian vein thrombosis is a dangerous complication of childbirth and often leads to inferior vena cava thrombosis and multiple pulmonary emboli. Computed tomography of the abdomen is useful in early diagnosis. Two patients with typical computed tomographic features are presented. PMID- 3052079 TI - Prospective study of carbohydrate metabolism in women using a triphasic oral contraceptive containing norethindrone and ethinyl estradiol for 3 months. AB - Thirty-five women with normal carbohydrate metabolism were administered a 3-hour oral glucose tolerance test before and after 3 months' use of a triphasic oral contraceptive that contained ethinyl estradiol and norethindrone. The results show no significant change in either the plasma glucose or the insulin values. This is the first published study with regard to this type of triphasic oral contraceptive, and the study supports claims of the preparation's improved safety. PMID- 3052081 TI - Comparison between lateral and axial ultrasonic measurements of the fetal femur. AB - The necessity of adequate determination of fetal age is well recognized, and several different parameters have been used for this purpose. Lately fetal femur length has been accepted as a relatively accurate determination. It is customary to measure the femur in the lateral plane; however, because of positional changes the femur sometimes must be measured vertically. The aim of our study was to perform lateral and axial measurements of the femur at various gestational ages and to examine the relationship between them. Our results reveal a significant difference between both measurements in all age groups. PMID- 3052082 TI - Sonographic appearance of the fetal heel ossification centers and foot length measurements provide independent markers for gestational age estimation. AB - The sonographic measurement of fetal foot length and the assessment of fetal heel ossification centers were conducted as an additional method for the estimation of gestational age. Such an approach is particularly desirable in cases where measurements of other biometric parameters are precluded, for example, in diseases such as anencephaly, hydrocephalus, and short limb dysplasia. A significant correlation was found between fetal foot length and gestational age (r = 0.9, p less than 0.0001) and between fetal foot length and femur length (r = 0.9, p less than 0.0001). Ossification centers of the calcaneus and talus were all present by the twenty-second week of gestation. However, their measurements discriminated between gestational ages of 18 to 22 weeks. The utilization of fetal foot length and heel ossification centers will serve as a useful adjunct in the estimation of gestational age and possibly in the diagnosis of diseases affecting the fetal foot. PMID- 3052083 TI - Reassessment of the utility of fetal umbilical vein diameter in the management of isoimmunization. AB - High-resolution ultrasound has been recently introduced in the management of many conditions in which the fetus is at risk. In the detection of severe erythroblastosis fetalis, sonographic evaluation of fetal anatomic changes is being used in association with amniotic fluid spectral analysis to assess the degree of the hemolytic process. In 1981 it was reported that sonographically detected umbilical vein dilatation, resulting from hepatic congestion, could be used as a sign of impending fetal compromise. We analyzed data obtained from 47 patients in a total of 76 examinations, including sonographic measurement of umbilical vein diameter, associated ultrasound findings, results of amniotic fluid spectral analyses, and neonatal outcome. The collected data, divided in two groups according to the results of amniotic fluid spectral analyses (less than high zone II and greater than or equal to high zone II), showed that the sonographic measurement of umbilical vein diameter does not differ significantly between the two groups (p greater than 0.05). Therefore, the present and relatively large series demonstrates that dimensions of the umbilical vein cannot be used as a reliable predictor of worsening fetal disease. PMID- 3052084 TI - Maternal and fetal blood flow velocity waveforms in patients with preterm labor: prediction of successful tocolysis. AB - Umbilical and uterine artery velocimetry was performed using continuous-wave Doppler ultrasound (Angioscan III) in 60 patients in preterm labor. Peak systolic/end-diastolic ratios were calculated according to previously described techniques. All measurements were made before tocolytic therapy was begun: magnesium sulfate (n = 40) or ritodrine (n = 20). The mean gestational age was 33.1 +/- 1.5 weeks (range 29 to 36 weeks). Twelve (20%) patients had elevated (greater than 2.6) pretherapy uterine peak-systolic/end-diastolic ratios, 10 (16.7%) patients had elevated (greater than 3.5) pretherapy umbilical peak systolic/end-diastolic ratios, and in eight (13.3%) patients both ratios were elevated. In seven (58.4%) of the 12 patients with elevated uterine peak systolic/end-diastolic ratios, six (60%) of the 10 patients with elevated umbilical peak-systolic/end-diastolic ratios, and five (62.5%) of the eight patients with both ratios elevated tocolytics failed and the women were delivered within 48 hours, compared with seven (14.6%) of 48, eight (16%) of 50, and six (13.0%) of 46 with normal ratios, respectively (p less than 0.05). We conclude that patients in preterm labor with elevated pretherapy uterine and/or umbilical peak-systolic/end-diastolic ratios are more likely to fail tocolysis therapy and be delivered preterm than those with normal ratios. It may therefore be useful to include umbilical and uterine velocimetry in the initial evaluation of preterm labor. PMID- 3052085 TI - Prenatal ultrasonographic recognition of Goldenhar's syndrome. AB - Goldenhar's syndrome is a series of malformations involving the face, either unilaterally or bilaterally. Other organs, such as the lungs, kidneys, spine, and heart, can also be involved. We report the prenatal ultrasonographic findings with regard to a fetus with multiple anomalies where Goldenhar's syndrome was not diagnosed until after birth. The importance of finding patterns of malformations is stressed to optimize the postnatal care for infants with multiple congenital abnormalities. PMID- 3052086 TI - Effects of prostacyclin on ovulation and microvasculature of the in vitro perfused rabbit ovary. AB - Prostacyclin (prostaglandin I2) methyl ester at 0.1, 1, or 10 micrograms/ml was added to the perfusate of one rabbit ovary every 2 hours for the first 10 hours of perfusion. The contralateral ovary was perfused with medium alone. Prostacyclin methyl ester at 1 and 10 micrograms/ml induced ovulation in the absence of gonadotropin, with ovulatory efficiencies of 26.7% +/- 3.8% and 46.5% +/- 5.0%, respectively. Most ovulated ova (77.4%) did not progress beyond the germinal vesicle stage, and there was no significant degeneration of ovulated ova or follicular oocytes. Examination of the follicular microvasculature 5 hours after exposure to prostacyclin revealed marked vessel dilatation and filling defects at the apex. By 7 hours after prostacyclin exposure, the intrafollicular space contained extravasated resin, reflecting increased vascular permeability. The vascular changes paralleled those previously observed in gonadotropin-induced ovulation. These results indicate that prostacyclin acting locally alters the vascular integrity of the follicle wall and facilitates follicle rupture. PMID- 3052087 TI - Adequacy of the ELISA test for screening corneal transplant donors. AB - Using a simple mathematical model, we calculated the risk for a patient undergoing penetrating keratoplasty to receive a cornea from a human immunodeficiency virus-infected donor despite negative results on serologic testing of donor serum. This error in serologic testing occurred when false negative results were obtained from the enzyme-linked immunosorbent assay used to screen donor corneas for human immunodeficiency virus exposure. The average risk of transplanting an infected cornea was low, 0.03%, but increased by a factor of ten when donor tissue from donors at high risk for AIDS was used. Current screening procedures are probably adequate to prevent transmission of human immunodeficiency virus, but increased vigilance for high-risk donor populations may be appropriate. PMID- 3052088 TI - Calibration of the Terry keratometer. PMID- 3052089 TI - The effect of suture removal on postkeratoplasty astigmatism. PMID- 3052090 TI - Presence of the Dr receptor in normal human tissues and its possible role in the pathogenesis of ascending urinary tract infection. AB - The Dr hemagglutinin of uropathogenic Escherichia coli recognizes the Dra blood group antigen, a component of the IFC or Cromer-related blood group complex. The present report used the Dr hemagglutinin to demonstrate location of the Dr receptor in selected human tissues and to evaluate the possible use of this lectin as a tissue marker recognizing sites sensitive for bacterial colonization. It was found that the Dr receptor was expressed in different parts of the digestive, urinary, genital, and respiratory tracts, and skin. Intense staining by Dr hemagglutinin was shown in colonic, bronchial, and endometrial glands, and skin eccrine sweat glands. Structures of the urinary tract showing strong fluorescence were renal tubular basement membrane, Bowmans' capsule, and transitional epithelium. The role of Dra antigen as receptor for adhesion for Dr positive E. coli in ascending colonization of urinary tract and the possible importance of Dra in human pathology is discussed. PMID- 3052091 TI - Mercuric chloride-induced autoimmunity in the brown Norway rat. Cellular kinetics and major histocompatibility complex antigen expression. AB - HgCl2 induces an autoimmune syndrome in Brown Norway rats that involves synthesis of anti-glomerular basement membrane (GBM) antibodies and development of nephritis with high proteinuria. HgCl2-induced changes in the composition of leukocyte populations and in the expression of MHC antigens in lymphoid and nonlymphoid organs were investigated by flow cytometry and immunohistochemistry. An early increase of CD4+ splenocytes was followed by a transient proliferation of CD4+ as well as CD8+ and B lymphocytes in peripheral lymphoid organs; in contrast, progressive depletion of the thymic cortex was found. B lymphocyte activation involved mainly the IgG1 and IgE isotypes. Nonlymphoid organs were infiltrated by MHC class II antigen expressing CD4+ and CD8+ T lymphocytes and monocytes; secondary to infiltration, mainly epithelial cells, being the main target of infiltrating cells, showed increased expression of MHC antigens. In glomeruli a 2.7-fold increase of CD8+ lymphocytes occurred after HgCl2 administration. The diverse autoimmune phenomena observed in this study fit with the hypothesized involvement of T lymphocytes autoreactive with MHC class II antigens. Apart from anti-GBM autoantibodies, a role for autoreactive CD8+ T lymphocytes must be considered in the pathogenesis of the HgCl2-induced autoimmune syndrome. PMID- 3052092 TI - Streptococcal cell wall-induced arthritis and flare-up reaction in mice induced by homologous or heterologous cell walls. AB - Intra-articular injection of cell walls from the gram-positive bacterium Streptococcus pyogenes induces an arthritis in both streptococcal cell wall (SCW) primed and naive mice. This joint inflammation subsides after 2 weeks but it could be reactivated by systemic injection of SCW in a dose-dependent way. The primary arthritis as well as the flare-up reaction were more vehement in immunized than naive mice. Pretreatment with antilymphocyte serum of nonimmunized arthritic mice before systemic challenge completely inhibits the flare-up reaction, suggesting the involvement of lymphocytes in the reactivation. Dose response studies showed that intravenous challenge with SCW amounts too small to induce a primary arthritis were able to reactivate a chronic arthritis, implying that an inflamed joint is in a hyperreactive state, probably due to locally retained lymphocytes. Arthritis as a result of injection with SCW can be reactivated by fragments of a nonrelated, gram negative endogenous bacterium, Escherichia coli. The latter finding might be of importance for the understanding of the pathogenesis of chronic arthritis: once an arthritis is induced by one bacterium, other (unrelated) bacteria, probably derived from an endogenous source, may be able to reactivate the inflammatory process, thus contributing to chronicity. PMID- 3052093 TI - Plasmacytoid T cells. Immunohistochemical evidence for their monocyte/macrophage origin. AB - To elucidate the lineage of plasmacytoid T cells, their immunophenotype was studied in reactive lymph nodes with a broad panel of monoclonal antibodies. Plasmacytoid T cells expressed several myelomonocytic markers, and almost all markers highly selective for macrophages. They lacked granulocyte-associated and B or T lymphocyte-associated antigens. These results provide strong evidence that plasmacytoid T cells are of monocyte lineage. PMID- 3052096 TI - CT and MRI of the ear and temporal bone: current state of the art and future prospects. AB - Computed tomography (CT) and magnetic resonance imaging (MRI) have become the radiographic investigations of choice for the vast majority of ear and temporal bone disorders. CT provides excellent images of bone and is indicated where osseous changes are of greatest diagnostic importance. MRI is superior in the evaluation of the eighth nerve complex and the central nervous system. Anticipated future advances include the production of three-dimensional images, more rapid scan times permitting dynamic (cine) studies, and the imaging of labeled chemical markers with insights into physiologic derangements. PMID- 3052094 TI - Intraglomerular deposition of fibrin/fibrinogen-related antigen in children with various renal diseases. AB - The localization of intraglomerular deposits of fibrin (Fb)/fibrinogen (Fg) related antigen (FRA) in children with various glomerular diseases was determined by an immunohistopathologic method using an anti-Fg antibody capable of detecting FRA, an anti-D-dimer antibody capable of detecting crosslinked Fb (XLFb) and its derivatives (XLFbDP), and by a method using the effect of monochloroacetic acid (MCA) treatment on kidney sections. In proliferative glomerulonephritis (PGN), XLFbs were detected within the capillaries and extension beyond the mesangium was seen in severe PGN. The FRA within the mesangium of minimal or mild PGN was composed of the non-XLFb substance. The FRA within Bowman's space of most PGN had disappeared after MCA treatment, suggesting a non-XLFb substance. The presence of FRA within electron-dense deposits (EDD) suggested that FRA deposits are associated with immune-complex deposits in the glomeruli. PMID- 3052095 TI - Early induction of MHC antigens in human liver grafts. An immunohistologic study. AB - The present study documents major histocompatibility complex (MHC) Class I and II expression during early acute rejection of human liver grafts. Serial graft biopsies (pretransplant, time zero, and 1 week) were studied. Ten patients received azathioprine (AZA) and prednisone; the other six patients were treated with quadruple therapy (azathioprine, cyclosporine A, prednisone, and cyclophosphamide). To study the specificity of changes in MHC antigen expression, biopsies of six patients with minor or no morphologic abnormalities served as controls. In addition, phenotypes of inflammatory cells present during rejection were analyzed using a panel of monoclonal antibodies. The results show that during acute rejection expression of MHC Class I and II antigens increased significantly in the AZA-treated patients, in a pattern similar to that seen in the patients treated with quadruple therapy, showing enhanced MHC Class I expression on hepatocytes, bile duct epithelium, and sinusoidal endothelium, and Class II antigen on Kupffer cells and sinusoidal endothelium. Bile duct epithelium was consistently positive for Class II antigen; no significant difference with the nonrejection group was observed. T cells are the predominant inflammatory cells during rejection with equal quantities of CD4+ and CD8+ cells. A majority of the infiltrating T cells show expression of Class II antigen but do not react with anti-interleukin-2 receptor antibody. This may be the result of immunosuppressive therapy or a simple reflection of the temporary expression of interleukin-2 receptors during lymphocyte activation. The authors hypothesize that the induction of MHC antigens on bile duct epithelium leads to rejection whereas the expression on hepatocytes represents an epiphenomenon. PMID- 3052097 TI - Vertigo in postoperative follow-up of otosclerosis. AB - Causes of vertigo after otosclerosis surgery were studied postoperatively and in long-term follow-up examinations. Pressure and mobility changes in the posterior labyrinth fluids, enzymatic process, and decrease in blood supply at the time of operation appear to be the major causes. Methods of detection, avoiding, and managing vertigo are presented. PMID- 3052098 TI - Intralabyrinthine schwannoma. AB - In the case presented, an intralabyrinthine schwannoma was discovered during translabyrinthine eighth nerve section. Analysis of this case and the 22 previously reported cases of intralabyrinthine schwannoma highlights the difficulty in preoperative diagnosis of this lesion. Concerns are raised about the frequency with which this diagnosis may be missed and its implication in neurotologic surgery for vertigo. PMID- 3052099 TI - Tympanoplasty and ossicular reconstruction: an update. PMID- 3052100 TI - Mammalian blastocyst: transport functions in a developing epithelium. AB - This article reviews the current state of knowledge concerning morphological and physiological mechanisms important to growth and differentiation of the mammalian blastocyst between compaction and implantation. Morphological processes occur in conjunction with major changes in transport systems that control the movement of substances into and out of the embryo. Compaction is a morphological development that is associated with the formation of an outer squamous epithelium, the trophectoderm, which regulates the composition of the medium bathing the presumptive embryo (the inner cell mass). Implantation involves the interaction of two epithelia, the adhesion between the trophectoderm and the maternal endometrium. Before adhesion, the blastocyst lies free in the uterine fluid and exchanges occur between this fluid and the embryo. Apposition of these epithelia is brought about in part by expansion of the blastocyst and removal of the uterine fluid. Blastocyst physiology is an inherently important field because vectorial transport system development and the genes that regulate it can be studied. PMID- 3052101 TI - Intracellular pathways of insulin transport across vascular endothelial cells. AB - Processing and transport of hormones across vascular endothelial cells may modulate hormone action at subendothelial tissue sites. Insulin was transported across cultured rat capillary and bovine aortic endothelial cells, after a delay of 5-10 min, at a constant rate for 60 min at 37 degrees C. 125I-labeled insulin transport was inhibited by 88 +/- 11% (SE, n = 4) and 75 +/- 18% (SE, n = 4) in the presence of anti-insulin receptor antibody and unlabeled insulin (at 10(-7) M), respectively. Reverse phase high-performance liquid chromatography showed 88% of the 125I-insulin transported over 60 min was indistinguishable from the 125I insulin added to the cells at 4 degrees C. In aortic endothelial cells preincubated with 2.3 x 10(-9) M of insulin for 24 h, insulin receptor binding was downregulated by 67%, and 125I-insulin transport was decreased by 52 +/- 11%. The proton ionophore monensin (0.05 mM) increased the internalized insulin in bovine aortic endothelial cells by 78%, with a corresponding decrease in 125I insulin released by 76 +/- 2% (SE, n = 4). 125I-insulin transport across the aortic endothelial cell monolayer was similarly decreased (54 +/- 12%, SE, n = 4) by monensin. In contrast, the lysosomal protease inhibitor leupeptin had no effect. Degradation and transport were similarly dissociated by low temperature. At 15 degrees C, no significant insulin degradation was detected, whereas 125I insulin release from the cells continued at 30 +/- 3% of the rate at 37 degrees C.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3052102 TI - Diazoxide unmasks glucose inhibition of insulin release by counteracting entry of Ca2+. AB - The interaction of diazoxide with the effects of glucose on the insulin-releasing mechanism was analyzed in beta-cell-rich pancreatic islets isolated from ob/ob mice. When added at a concentration of 400 microM to a medium containing 1.28 mM Ca2+, diazoxide converted glucose stimulation of insulin release into inhibition. Further addition of 2 mM theophylline restored the insulin secretory response to glucose. The paradoxical glucose inhibition of insulin release was accounted for by a diazoxide interaction with the entry of Ca2+, unmasking a capacity of the sugar to lower cytoplasmic Ca2+ below its resting concentration. PMID- 3052103 TI - Metabolic role of the exercise-induced increment in epinephrine in the dog. AB - The role of the exercise-induced increment in epinephrine was studied in five adrenalectomized (ADX) and in six normal dogs (C). Experiments consisted of an 80 min equilibration period, a 40-min basal period, and a 150-min exercise period. ADX were studied with epinephrine replaced to basal levels during rest and to increased levels during exercise to simulate its normal rise (HE) and on a separate day with epinephrine maintained at basal levels throughout the study (BE). Cortisol was replaced during rest and exercise in ADX so as to simulate the levels seen in C. Glucose was infused as needed in ADX to maintain the glycemia evident during exercise in C. Glucose production (Ra) and utilization (Rd) were assessed isotopically. In C, epinephrine had risen by 95 +/- 25 pg/ml by the end of exercise. In HE, the increment in epinephrine (117 +/- 29 pg/ml) was similar to that seen in C, whereas in BE epinephrine fell by 18 +/- 9 pg/ml. Basal norepinephrine levels were 139 +/- 9, 260 +/- 25, and 313 +/- 33 pg/ml in C, HE, and BE, respectively. In response to exercise, norepinephrine increased by nearly twofold in all protocols. Basal and exercise-induced changes in plasma glucagon and insulin were similar in C and ADX. Ra increased similarly in C (5.3 +/- 0.6 mg.kg-1.min-1) and HE (4.9 +/- 0.6 mg.kg-1.min-1). In BE, Ra rose normally for the initial 90 min but then declined resulting in a rise of only 2.9 +/- 0.5 mg.kg-1.min-1 after 150 min of exercise.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3052104 TI - Multiple renin forms in the adrenal gland. AB - Renin heterogeneity has been described in rat kidney and plasma. In this study, we used the isoelectric focusing method to 1) characterize the adrenal renin forms in control rats, in rats on low- and high-Na diets, and in nephrectomized rats; and 2) examine their resemblance with plasma renin. Active renin (AR) and inactive trypsin-activatable renin (IR) were measured in adrenal homogenates and plasma. Aliquots were subjected to isoelectric focusing gels. Activation with trypsin (5 mg/ml) was performed before or after isoelectric focusing. Results showed that adrenal glands contained AR and IR. The content of adrenal AR increased significantly only in rats fed a low-Na diet. Following anesthesia, nephrectomy, or high-Na intake, the content of adrenal AR and IR was not significantly changed. In plasma, an inverse relationship between AR and IR was found. Adrenal glands contained six forms of AR focusing at the same pH as those of plasma AR but in different proportions. After activation of IR in adrenal glands, two additional renin forms focusing at pH 6.4 and 6.1 were found, whereas after activation of plasma IR, two peaks focusing at pH 5.9 and 4.8 were significantly enhanced. Adrenal AR forms were modified by alterations of salt and water balance differently than plasma AR. These results support the hypotheses of an endogenous production of renin forms by the adrenal gland. PMID- 3052105 TI - Acute inhibition of rat heart protein synthesis in vitro during beta-adrenergic stimulation or hypoxia. AB - In the anterogradely perfused rat heart with glucose as fuel, 1 microM isoproterenol (ISO) inhibited the insulin (INS) plus adenosine deaminase (AdoDA) stimulation of ventricular protein synthesis by 72%. ISO (1 microM) alone had no effect on ventricular protein synthesis but inhibited atrial protein synthesis by 20%. The concentration dependence of the ISO inhibition was similar to the stimulation of glucose uptake by ISO. Inhibition could not be overcome by increasing INS concentrations. The effects of ISO were diminished by propranolol and could be partially mimicked by forskolin (FSK) or 8-(4-chlorophenylthio )adenosine 3',5'-cyclic monophosphate (CPT-cAMP). The stimulation of protein synthesis by noncarbohydrate fuels was antagonized by ISO. Hypoxia (PO2 = 50%) also antagonized the INS stimulation of ventricular protein synthesis but did not affect basal rates. ATP contents were decreased by ISO but not by a PO2 of 50%. Both manipulations increased lactate output. The inhibition of protein synthesis by ISO could possibly be explained by indirect effects of ISO on cardiac "energy status." Furthermore, inhibition may thus represent purely an in vitro phenomenon and may not occur in vivo. However, the possibility that there are more direct effects of ISO on the machinery of protein synthesis has not been excluded. The inhibition of protein synthesis by hypoxia cannot be explained by changes in energy status and may result from intracellular lactoacidosis. PMID- 3052106 TI - Insulin responsiveness of protein metabolism in vivo following bedrest in humans. AB - To test the influence of bedrest on insulin regulation of leucine metabolism, six normal young men were subjected to a five-step hyperinsulinemic euglycemic clamp before and after 7 days of strict bedrest. A primed-constant infusion of [1 13C]leucine at 0.12 +/- 0.02 mumol.kg-1.min-1 was used. Before bedrest, the basal rate of appearance (Ra) of intracellular leucine and leucine oxidation were 2.79 +/- 0.17 and 0.613 +/- 0.070 mumol.kg-1.min-1, respectively. Insulin caused a dose-dependent reduction of the intracellular leucine Ra and leucine oxidation to a minimum of 1.64 +/- 0.08 and 0.322 +/- 0.039 mumol.kg-1.min-1, respectively, in nonbedrested subjects (P less than 0.001). Insulin also caused a dose-dependent reduction of plasma leucine concentration from 95 +/- 4 to 38 +/- 2 mumol/l (P less than 0.001). After bedrest, subjects exhibited decreased glucose tolerance and increased endogenous insulin secretion, but basal and insulin-suppressed intracellular leucine Ra and leucine oxidation rates were not different from control. Magnetic resonance imaging of the back and lower extremities revealed a 1-4% decrease in muscle volume and a 2-5% increase in fat volume secondary to bedrest. Bedrest also resulted in a negative nitrogen balance as compared with the control period, with an average cumulative loss of 6.3 g of nitrogen after 6 days. Urinary 3-methyl-L-histidine excretion was unchanged by bed rest. Thus because negative nitrogen balance and skeletal muscle atrophy occurred in six rested subjects in the absence of changes in the two indices of protein breakdown used in this study (3-methyl-L-histidine release and leucine release), it seems likely that muscle protein synthesis was inhibited. PMID- 3052107 TI - Impact of sex steroids and their suppression on skeletal growth and maturation. AB - Forty girls with central precocious puberty (CPP) were studied before and during 1-3 yr of luteinizing hormone-releasing factor (LHRH) agonist (LHRHa) administration to examine the impact of gonadal steroid secretion and its suppression on skeletal growth and maturation. Pubertal growth velocity (GV) was 10.1 +/- 0.7 (SE) cm/yr and, when normalized for chronological age (CA) and bone age (BA), demonstrated that the effects of sex steroids were most profound in patients with the youngest CA and BA. GV decreased significantly to 5.8 +/- 0.3 (n = 40), 4.6 +/- 0.3 (n = 30), and 3.2 +/- 0.6 cm/yr (n = 12) during 3 yr of gonadal suppression and correlated negatively with starting BA. Skeletal maturation was markedly accelerated by premature sex steroid secretion (BA/CA = 1.8 +/- 0.1), was slowed significantly with gonadal suppression (mean delta BA/delta CA less than 1), and also was negatively correlated with the starting BA. Cumulative increases in predicted adult height were observed regardless of starting BA and averaged +2.0 +/- 0.4, +5.2 +/- 0.5, and +6.7 +/- 1.2 cm after 1, 2, and 3 yr of gonadal suppression. The comparable changes in height predictions across all BAs despite highly variable GVs underscore the need for use of developmental (i.e., BA-based) rather than CA-based standards in the analysis of growth during gonadal steroid exposure and suppression in childhood. PMID- 3052108 TI - Experimental models of gastric ulceration and injury. AB - There are inumerable experimental models of gastric mucosal epithelial injury. Many of these currently in wide use can be regarded as unphysiological and severe, rarely encountered in humans. An analysis of more physiological and simple models indicates that little is known of the events that ultimately cause cellular death, even in simple and easily controllable systems. A review of acceptable measures of gastric mucosal injury is presented. It is suggested that studies of subtle physiological injuries at the cellular level are more likely to yield meaningful insights into the causation of gastric mucosal ulceration. PMID- 3052109 TI - Molecular biology of the renin-angiotensin system. AB - This paper reviews the molecular biology of the renin-angiotensin system. The renin gene structure is analyzed in detail, including an examination of the putative regulatory regions. The combined action of these regulatory sequences would result in the complex, tissue-specific expression and regulation observed in vivo. The expression of the tissue renin-angiotensin systems, which may have important physiological functions, is also described. In addition, the pathway of renin biosynthesis and secretion is reviewed. This includes speculation on the fate of circulating prorenin and the physiological role of multiple renin forms and secretory pathways. The molecular approaches described in this paper have greatly advanced our knowledge of the biology of the renin-angiotensin system. Future studies using these and other approaches should provide further insight into this complex system. PMID- 3052110 TI - Stimulatory effects of neuronally released norepinephrine on renin release in vitro. AB - Extracellular high potassium inhibits renin release in vitro by increasing calcium concentrations in the juxtaglomerular cells. We found that the decreased response of renin release from rat kidney cortical slices in high potassium solution (20-80 mM) changed to a strikingly increased one in the presence of nifedipine at doses over 10(-6) M. We then examined the stimulatory effect of extracellular high potassium in the presence of nifedipine on renin release. The enhancement of release was significantly suppressed either by propranolol or by metoprolol but not by prazosin. High potassium plus nifedipine-induced increase in renin release was markedly attenuated by renal denervation. The enhancing effect was not observed when the slices were incubated in calcium-free medium. Divalent cations such as Cd2+, Co2+, and Mn2+ (0.1-3.0 mM) blocked this enhancement in a concentration-dependent manner. High potassium elicited an increase in 3H efflux from the slices preloaded with [3H]norepinephrine. The increasing effect was not influenced by nifedipine but was abolished by the removal of extracellular calcium or by the addition of divalent cations. These observations suggest to us that the high potassium plus nifedipine-induced increase in renin release from the slices is mediated by norepinephrine derived from renal sympathetic nerves and that this neuronally released norepinephrine stimulates renin release via activation of beta-adrenoceptors. PMID- 3052111 TI - Intrarenal renin inhibition increases renal function by an angiotensin II dependent mechanism. AB - ACRIP is a competitive inhibitor of renin in which an analogue of statine, (3R,4S)-4-amino-3-hydroxy-6-methylheptanoic acid, is incorporated into analogues of porcine renin substrate. ACRIP inhibits the enzymatic activity of renin, thus blocking the initiation of the angiotensin cascade. We studied the intrarenal action of ACRIP in small quantities without measurable systemic effects on renal function. In the first experiment, ACRIP was administered intrarenally at 0.02, 0.2, and 2 micrograms.kg-1.min-1 to uninephrectomized conscious dogs (n = 6) in metabolic balance at sodium intake of 10 meq/day. ACRIP, in doses of 0.02 and 0.2 micrograms.kg-1.min-1, markedly increased urine sodium excretion (UNaV) from 5.8 +/- 1.4 to 15.1 +/- 5.1 and 19.9 +/- 3.2 mu eq/min, respectively. Urinary flow rate (UV) underwent a similar increase and glomerular filtration rate (GFR) increased from 25.7 +/- 2.5 to 35.6 +/- 2.5 at 0.02 micrograms.kg-1.min-1 of ACRIP. Renal plasma flow (RPF), plasma renin activity (PRA), and plasma aldosterone concentration (PAC) were not affected. At 2 micrograms.kg-1.min-1, ACRIP traversed the kidney in quantities large enough to produce a reduction in systemic PRA and mean arterial pressure and caused natriuresis, diuresis, and increased GFR. In a second experiment, ACRIP was administered intrarenally at 0.2 micrograms.kg-1.min-1 in a separate group (n = 4) under identical conditions. ACRIP-induced increases in UV and UNaV were completely blocked by concurrent intrarenal administration of angiotensin II. The results indicate that intrarenal angiotensin II acts as a physiological regulator of renal sodium and fluid homeostasis. PMID- 3052112 TI - Captopril augments the renal response to an amino acid infusion in diabetic adults. AB - This study examined whether patients with insulin-dependent diabetes mellitus and normal renal function have an altered response to an amino acid infusion when they are pretreated with a converting-enzyme inhibitor. Three groups of adults received amino acid infusions for 20 min on two occasions separated by a 240-min interval. Groups 1 (6 normals) and 2 (6 diabetics) ingested captopril (12.5 mg) 120 min before starting the second infusion. Group 3 (4 diabetics) did not receive captopril. Diabetics had normal base-line renal plasma flow, as indicated by para-aminohippuric acid clearance (CPAH), and glomerular filtration rate (GFR). In group 1, the maximum increase in CPAH was significant and similar with both infusions, 23 +/- 5 vs. 15 +/- 3% (SE); maximal changes in GFR were also significant and similar, 20 +/- 5 vs. 20 +/- 6%. In Group 2, the maximal increase in CPAH and GFR with the first infusion was 28 +/- 7 and 23 +/- 6%, respectively. After captopril, the increases in CPAH and GFR were significantly greater than with the first infusion (64 +/- 8%, P less than 0.002, and 67 +/- 9%, P less than 0.002, respectively). In Group 3 diabetics, there was no difference in either CPAH or GFR with the first vs. the second infusion. Thus captopril enhances the renal hemodynamic response to an amino acid load in diabetic patients but not in normal adults. PMID- 3052113 TI - Renal hemodynamics and vascular reactivity in canine DOCA-salt hypertension. AB - Because of the potential role that the kidney may play in deoxycorticosterone acetate (DOCA)-salt hypertension, changes in renal blood flow, renal vascular reactivity, and renal adrenergic vascular tone were followed in the conscious instrumented dog. DOCA-salt was administered daily after obtaining control measurements. Systemic mean arterial blood pressure (MAP) was monitored with an indwelling catheter, renal blood flow (RBF) was measured electromagnetically using an implanted blood flow probe, and drugs were administered intrarenal arterially through an indwelling renal artery catheter. During the first week of DOCA-salt administration MAP increased from 106 +/- 3 to 118 +/- 2 mmHg and at week 2 to 123 +/- 2 mmHg (P less than 0.01). RBF increased from 275 +/- 32 to 336 +/- 34 during week 1 (P less than 0.05) and to 324 +/- 29 ml/min during week 2. The log ED50 of norepinephrine administered intra-arterially decreased from 1.66 +/- 0.114 to 1.48 +/- 0.091 and 1.41 +/- 0.067 ng/ml (P less than 0.05), and of angiotensin II from 2.58 +/- 0.072 to 2.31 +/- 0.09 (P less than 0.05) and 2.38 +/- 0.05 pg/ml, during weeks 1 and 2, respectively. There was, however, no increase in adrenergic vascular tone as determined by the change in RBF obtained with the intra-arterial infusion of alpha-adrenoceptor antagonists. These experiments indicate that RBF is not compromised in canine DOCA-salt hypertension, and renal adrenergic tone is no greater in the hypertensive than in the normotensive control period. PMID- 3052114 TI - Physical activity and coronary heart disease among asymptomatic hypercholesterolemic men (the Lipid Research Clinics Coronary Primary Prevention Trial). AB - We examined the relation between reported regular strenuous exercise or hard physical labor and the incidence of coronary heart disease (CHD) death and nonfatal myocardial infarction among 1,533 hypercholesterolemic men aged 35-59 years who were in the placebo group of the Lipid Research Clinics Coronary Primary Prevention Trial. The mean follow-up of the cohort was 7.4 years. The men were free of clinical heart disease at entry; men with an abnormal resting electrocardiogram or graded exercise test also were excluded. Regular physical activity was not associated with the incidence of CHD (RR = .94, 95% CI = .68, 1.38) in this study population. Adjustment by the proportional hazards model for age, low-density lipoprotein cholesterol, smoking, family history of CHD, and occupation did not alter this finding. This observation suggests that reported regular physical activity may not be related to the risk of coronary heart disease among asymptomatic, hypercholesterolemic, middle-aged men. PMID- 3052115 TI - The persistence of Shigella flexneri in the United States: increasing role of adult males. AB - The annual reported isolation rate of Shigella flexneri decreased from 1964 to 1973, but has remained constant since then at 1 per 100,000. Between 1975 and 1985, the median age of males from whom S. flexneri was isolated rose from 5 to 26 years. During this time, the isolation rate of S. flexneri rose more than five fold among men, did not change in adult women, and decreased in children. By 1985, 23 per cent of reported S. flexneri isolates came from men aged 20-49. Increased male homosexual transmission of S. flexneri is a possible explanation for these findings. PMID- 3052117 TI - A comparative field study of radiolabeled and enzyme-conjugated synthetic DNA probes for the diagnosis of falciparum malaria. AB - Synthetic, 21-base, DNA probes to the genome of Plasmodium falciparum were either 32P-labeled or enzyme-conjugated for comparative field studies. The sensitivity of both probes was compared with microscopy in the examination of blood samples from 97 Thai villagers, 47 Thai Rangers, and 19 malaria-free Bangkok residents. The probes were also used to monitor the therapeutic response of 18 of the Rangers during 7 days of treatment. The probes proved highly specific. Both probes had lower limits of detection of about 100 parasites per microliter blood. Thus, the low parasite densities in partially immune villagers from an endemic area were often missed, while higher parasite densities in the nonimmune Rangers were usually detected. As monitors of response to treatment, the probes paralleled microscopy in identifying reversion from positive to negative parasitemia. The enzymelabeled DNA probe as shown to perform similarly to the radiolabeled probe in populations with different malarial immune status and during curative treatment. PMID- 3052116 TI - Improving the life-course development of socially disadvantaged mothers: a randomized trial of nurse home visitation. AB - We evaluated a comprehensive program of prenatal and postpartum nurse home visitation for socially disadvantaged women bearing first children. Eighty-five per cent of the participating women were either teenagers (less than 19 years at registration), unmarried, or of low socioeconomic status. Women were randomly assigned to either nurse home visitation or comparison services (free transportation for prenatal and well-child care and/or sensory and developmental screening for the child). During the first four years after delivery of their first child, in contrast to their counterparts in the comparison group, nurse visited White women who had not graduated from high school when they registered in the study returned to school more rapidly; nurse-visited, poor, unmarried White women showed an 82 per cent increase in the number of months they were employed, had 43 per cent fewer subsequent pregnancies, and postponed the birth of second children an average of 12 months longer. PMID- 3052118 TI - Iron chelators: in vitro inhibitory effect on the liver stage of rodent and human malaria. AB - The activity of desferrioxamine (Desferal) and desferrithiocin (a newly developed oral iron chelator) was evaluated against the liver stage of Plasmodium yoelii and P. falciparum in the rodent and the human hepatocyte in vitro culture system. The two iron chelators were found to inhibit the liver schizogony of both the rodent and the human Plasmodium species at concentrations achievable in vivo. P. falciparum proved to be more sensitive (ic 95% below 20 micromol/l than P. yoelii (ic 95% 50-100 micromol/l). As assessed by electron microscopy, drug administration was associated with focal clarification of the cytoplasm thought to be reversible. As desferrioxamine and desferrithiocin are known to be equally active on the blood stage of rodent and human plasmodia, iron chelators are deserving of further investigation as potential alternative candidates to existing drugs for radical cure of malaria. PMID- 3052120 TI - Enzyme immunoassay in cell monolayers for evaluation of in vitro activity of chemotherapeutic agents against Trypanosoma cruzi. AB - A standardized enzyme-linked immunosorbent assay (ELISA), performed directly on monolayers of mouse peritoneal macrophages or L 929 fibroblasts, was used to evaluate the activity of chemotherapeutic agents against four different stocks of Trypanosoma cruzi. Absorbance readings, performed in an automatic ELISA reader, were directly related to the number of intracellular parasites as determined by microscopic examination of tissue culture slides run in parallel. Results were highly reproducible in replicate wells and in repeated experiments. Results with nifurtimox, ketoconazole, and formycin B, compounds known to have in vivo activity against T. cruzi, revealed that the ELISA technique was capable of detecting small dose-response variations in the rate of phagocytosis of different life cycle stages of T. cruzi by normal and activated mouse macrophages. PMID- 3052121 TI - Our legacy of thyroid surgery. PMID- 3052119 TI - Antibody responses to malarial antigens in the Wopkaimin population of the Star Mountains, Papua New Guinea. AB - Antibody responses to malarial antigens were determined in 614 serum samples collected from the Wopkaimin population of the Star Mountains of Papua New Guinea. In point prevalence surveys made in 1982-1983, 33.7% of the persons examined were infected with Plasmodium falciparum, P. vivax, or P. malariae. Of these, 72.9% were infected with P. falciparum. In a standard fluorescent antibody test, highest level responses were to P. falciparum, followed by P. malariae, P. vivax, and P. ovale. A strong correlation was found between results of the fluorescent antibody tests and those obtained in an enzyme-linked immunosorbent assay using P. falciparum antigens. The failure of immune responses to eliminate these species of Plasmodium in this highly isolated population is discussed. PMID- 3052122 TI - Enhancement of rehabilitation by use of implantable adjuncts with vascularized bone grafts for mandible reconstruction. AB - Restoration of mandibular continuity after trauma or cancer by means of vascularized bone grafts, as described by Taylor and coworkers in 1979, has become an established method of oral rehabilitation. Still considered by some an endpoint in reconstruction, we have found that the judicious application of newly described dental prosthetic materials can provide the patient with improved appearance and function associated with a fully restored mandible. Twelve patients who underwent vascularized bone grafting for mandibular reconstruction had adjunctive implantation to improve either dentition or to reconstruct the temporomandibular joint. Nine patients had dental implants to restore permanent stable dentition either by placement of the mandibular bone staple plate or osseointegrated implants. Three additional patients underwent placement of metallic condylar head prostheses placed on the vascularized bone graft at the time of transfer. These patients demonstrated good mastication and an excellent incisal opening which was maintained in the late postoperative period. Vascularized bone transfer for restoration of continuity of the mandible should not be considered an endpoint in and of itself. By using implantable dental devices to restore dentition and condylar head prostheses to improve both the aesthetic result and function of the mandible to complement the vascularized osteocutaneous flap, we can bring our patients closer to the goal of total rehabilitation. The majority of these patients would otherwise never have been offered a chance at restoration of dentition. All had beneficial effects with regard to mastication and the majority also had improved speech. PMID- 3052123 TI - Preoperative localization of parathyroid adenomas. AB - During a 12-month period, 64 patients were operated on for primary hyperparathyroidism. Sixty-one had single adenomas and 3 had double adenomas. Preoperative imaging was used to localize the adenomas. Half of the patients (32 of 64) had magnetic resonance, thallium-201/technetium-99m subtraction scintigraphy, and high-resolution ultrasonography; the other 32 patients had 1 or 2 of the imaging modalities. Sensitivity and specificity of magnetic resonance imaging was 82 percent and 97 percent, respectively; the sensitivity and specificity of the other two modalities was 59 and 98 percent for subtraction scintigraphy and 73 and 98 percent for ultrasonography. The use of preoperative imaging facilitated surgical exploration, reduced operating time, and resulted in an increased number of successful operations. There were no negative explorations in this series as compared with 19 negative explorations (2.6 percent) in our prior experience with 720 operations. PMID- 3052124 TI - [Echographic characteristics of endometrial cancer]. PMID- 3052125 TI - [Cyanoacrylate glues and their use in gynecology]. PMID- 3052126 TI - [Surgical correction of ptosis of the vaginal walls during uterine extirpation]. PMID- 3052127 TI - [Elongation of the cervix uteri and its surgical treatment]. PMID- 3052128 TI - [Echographic diagnosis of inflammatory diseases of the adnexa uteri]. PMID- 3052129 TI - [Clinical laboratory research on habitual abortions of presumed immune origin]. PMID- 3052130 TI - [Changes in the size of fetal kidneys determined by ultrasonics in advanced pregnancy]. PMID- 3052131 TI - [Perinatal outcome in pregnant women with diabetes mellitus]. PMID- 3052132 TI - [Use of echography in the diagnosis of ovarian tumors]. PMID- 3052133 TI - [Resuscitation problems in pregnant women with pre-eclampsia]. PMID- 3052134 TI - [Antiprogestins--the possibility of a new type of contraceptives]. PMID- 3052135 TI - Orphan drugs. Improved medical treatment of rare diseases. PMID- 3052136 TI - Anaphylaxis. Part I. Etiology and pathogenesis. PMID- 3052137 TI - Transmethylation in depression. PMID- 3052138 TI - An open-label pilot study of oral S-adenosylmethionine in major depression. An interim report. PMID- 3052139 TI - S-adenosylmethionine in depression. A literature review and preliminary report. PMID- 3052140 TI - Rapid antidepressant response with SAMe. A double-blind study. PMID- 3052141 TI - Switch and S-adenosylmethionine. PMID- 3052142 TI - Procurement of cadaver kidneys for transplantation. A joint university/community effort. PMID- 3052144 TI - Regulation of nausea and vomiting in cancer chemotherapy. A review with emphasis on opiate mediators. AB - Chemotherapy-induced nausea and vomiting are primarily regulated by chemoreceptor trigger zone (CTZ)-vomiting center (VC) pathways. Dopaminergic (D2), histaminic (H1), and muscarinic cholinergic (Ach) receptors are present in these sites, and specific receptor antagonists are potent but not "universal" antiemetics when used alone or in combination. Recently, neurons containing the endogenous opiate enkephalin were also identified near the CTZ and the VC. Furthermore, opiates stimulate vomiting at the CTZ and inhibit vomiting at the VC in dogs and in cats. A dose-related increase in nausea and vomiting in response to the opiate antagonist naloxone has also been demonstrated in patients receiving cancer chemotherapy. These observations support a role for endogenous opiates in regulating chemotherapy-induced nausea and vomiting; further, they suggest that narcotic agents may be effective antiemetics in this setting. PMID- 3052143 TI - Increased nausea and vomiting induced by naloxone in patients receiving cancer chemotherapy. AB - To evaluate the role of endogenous opiates in chemo-therapy-induced nausea and vomiting, the narcotic-antagonist naloxone was administered to six pediatric patients receiving cancer chemotherapy. Naloxone was administered by continuous intravenous (i.v.) infusion after randomized, double-blind controlled assignment at a dose of 0, 10 or 40 micrograms/kg/h for 12 h. Each patient was studied for four consecutive and identical courses of chemotherapy (eight courses for each naloxone dose or 24 courses in all). A dose-related increase in nausea (nausea score 2.5 +/- 2.24, 3.83 +/- 2.73, and 5.75 +/- 2.86/12 h, p = 0.003), vomiting (emetic events 6.0 +/- 7.50, 8.08 +/- 6.71, and 10.3 +/- 8.91/12 h, p = 0.035), and patient aversion (course preference rank 1.5 +/- 0.45, 2.83 +/- 1.17, and 3.25 +/- 0.42/4 courses, p = 0.014) was observed. The infusion of naloxone in the absence of chemotherapy was without effect. These results support a role for endogenous opiates in regulating chemotherapy-induced nausea and vomiting, and further suggest that narcotic agents may be effective antiemetics in this setting. PMID- 3052145 TI - Does stress affect bleeding in hemophilia? A review of the literature. AB - Many bleeding episodes in hemophilia are thought to be related to ambient stress. The evidence in support of this hypothesis comes from a variety of anecdotal and clinical reports. Whether or not stress leads to bleeding has had little direct study in a prospective way, however, and methodological problems affect most of the studies in this field. This paper examines the evidence in support of this hypothesis and suggests ways to improve research relating stress to bleeding. PMID- 3052146 TI - Neonatal hemophilia B with intracranial hemorrhage. Case report. AB - It is uncommon for infants with hemophilia to have excessive bleeding during the neonatal period. Even if bleeding occurs, it rarely becomes life-threatening, such as in intracranial hemorrhage (ICH). We here report a case of a 4-day-old boy who had intracranial hemorrhage as the first complication of hemophilia B. Computerized axial tomography (CT scan) and ultrasonography were very useful for early diagnosis. Only a few cases of neonatal hemophilia with intracranial hemorrhage have been reported, but the occurrence of this complication is probably more frequent. We reviewed seven cases (including our case) with intracranial hemorrhage as the first manifestation of neonatal hemophilia. Although these infants showed good prognosis as to survival, permanent residual neurological deficits remained in all of them. It is emphasized that intracranial hemorrhage due to hemophilia may occur in neonates even without a family history. Urgent neuroimaging and coagulation studies are necessary for an early and adequate diagnosis. PMID- 3052147 TI - Juvenile chronic myelogenous leukemia. AB - Juvenile chronic myelogenous leukemia (JCML) is a malignant hematopoietic disorder of monocyte-histiocyte lineage that affects children less than 4 years of age. Since the disease represents only 2% of all childhood leukemias, experience with it has been limited even in large centers. This review summarizes our 10 year institutional study of JCML as well as a comprehensive literature survey. The goal of the article is to underscore the cardinal features of juvenile chronic myelogenous leukemia that are useful for diagnosis, and to highlight recent advances in the understanding of the biology and treatment of the disease. PMID- 3052148 TI - Electronic information for physicians: a new dimension in solving traditional problems. Part 1. PMID- 3052149 TI - History of medicine in Alaska. Jean Colesberry Persons, M.D. PMID- 3052150 TI - Glimpses of Alaskan medical history. A primer of Aleut medicine. PMID- 3052151 TI - Immunotherapy. Immunotherapy Subcommittee of the European Academy of Allergology and Clinical Immunology. PMID- 3052152 TI - [AIDS and anesthesia]. AB - AIDS will become a bigger and bigger problem in future, especially for medical staff who will be increasingly in contact with infected patients. The epidemiology of HIV infection (blood, vaginal secretion, sperm) and the threat of this infection require particularly strict observance of hygienic measures. Unqualified handling of infected material such as HIV-contaminated injection needles or pointed objects represent a major risk of HIV infection for medical personnel. Despite the high degree of safety in the preparation of banked blood we cannot completely guarantee that only absolutely safe HIV-free blood will be transfused. Hence, indication for transfusion must be very strictly limited. Autologous transfusion as a safe alternative to homologous transfusion should be employed more frequently. Seroconversion rate after needle-point injury is now stated to be one per cent. According to Goebel et al. in AIFO 5:227 (1988) four nurses carried out mouth-to-mouth resuscitation in an AIDS patient who had jumped from the third floor of a building in attempted suicide. Despite considerable blood contact the HIV antibody tests remained negative even now after 18 months. PMID- 3052153 TI - [Pharmacologic modification of vigilance in the postnarcotic phase-- naloxone or physostigmine?]. AB - The usefulness of physostigmine in reversing post-narcotic depression after general anaesthesia is well proven; so is that of naloxone, a specific opioid analgetics antagonist, in reversing neuroleptic anaesthesia effects. Morphine like analgetics are widely used as premedication agents, too; on the other hand, physostigmine reverses opioids as well as other psychotropic and narcotic agents. For that reason, positive post-narcotic physostigmine effects could be due to its anti-opioid potency as well. In a double-blind, randomised study, physostigmine and naloxone were evaluated using a clinically based vigilance protocol, and compared with saline solution. Naloxone did not have remarkable advantages as compared with placebo, while physostigmine led to a significantly higher level of vigilance; moreover, that level was reached sooner. The positive effects of physostigmine in restoring a sufficient level of vigilance after general anaesthesia are, in respect of our findings, unrelated to its antagonism to morphine-like analgetics. PMID- 3052155 TI - [Results of the analytic and sociometric study of books and articles on otology. 16th-20th centuries (up to 1932). 2]. PMID- 3052154 TI - [Critical comments on reports of fatalities in hereditary fructose intolerance in adulthood from the viewpoint of neuroanesthesia]. AB - In view of repeated communications in recent years reporting on lethal infusions of fructose or sorbitol in adults with hereditary fructose intolerance, the known statements on the incidence of 1:20,000 are critically analysed. The validity is relativated. The special indication for sorbitol as an osmotherapeutic preparation for lowering intracranial pressure is pointed out. A modified intravenous fructose tolerance test is suggested. PMID- 3052156 TI - [Tinnitus]. PMID- 3052157 TI - [Results of the analytic and sociometric study of books and articles on otology. 16th to 20th centuries (up to 1932). Part 3]. PMID- 3052158 TI - Directory of accredited organizations, approved programs/offerings, and accredited continuing education certificate programs preparing nurse practitioners: Spring 1988. PMID- 3052159 TI - The antigravity suit in neurosurgery. Cardiovascular responses in seated neurosurgical patients. AB - The haemodynamic responses associated with inflation of the antigravity suit (G suit, aviation type) to 8.0 kPa were studied in a series of 40 patients who underwent neurosurgical operations in the sitting position. The study showed statistically significant increases in systolic arterial pressure (p less than 0.005) and mean central venous pressure (p less than 0.001) with inflation of the suit. The systolic arterial and mean central venous pressures remained significantly elevated immediately before deflation of the suit at the end of the operation (p less than 0.001 and p less than 0.005 respectively). The addition of 0.8-1.0 kPa positive end expiratory pressure during suit inflation was also investigated. A further increase in central venous pressure occurred but this did not achieve statistical significance. PMID- 3052160 TI - Patient-controlled analgesia--the need for caution. A case report and review of adverse incidents. AB - Respiratory depression and oversedation occurred during the administration of patient-controlled analgesia because of a broken syringe barrel. Possible complications in the use of patient-controlled analgesia are reviewed. These may be because of prescription or provision errors, patient factors or equipment problems. PMID- 3052161 TI - The development of the anaesthetic vaporizer. The contribution of A.G. Levy. AB - The innovatory features incorporated into A.G. Levy's regulating chloroform inhaler, and their contribution to modern vaporizer design, are examined. Levy may be credited with the important realisation that the gas mixture which leaves the vaporizing chamber is fully saturated at all flow rates; with the concept of the splitting ratio; with appreciation of the importance of the application of Reynolds' researches on laminar and turbulent flow to the design of anaesthetic apparatus; and with the provision of a facility to compensate for changes of temperature in the vaporizing chamber. PMID- 3052162 TI - The first use of physostigmine in the treatment of atropine poisoning. A translation of Kleinwachter's paper entitled 'Observations on the effect of Calabar bean extract as an antidote to atropine poisoning'. PMID- 3052163 TI - Early textbooks on anaesthesia. PMID- 3052164 TI - [Alfred Doenicke on his 60th birthday]. PMID- 3052165 TI - [Anesthesia and intraocular pressure]. AB - General anesthesia has been in use for ophthalmic surgery since 1847. The subsequent predominance of local anesthetic techniques made ophthalmic anesthesia the "Cinderella of anesthesia services" until its clinical and scientific rehabilitation in the second half of this century. Precise control of intraocular tension is an accepted advantage of general anesthesia. The exercise of such control requires understanding of intraocular physiology and the effects exerted by anesthetic techniques. Hence, the impact of anesthetic drugs on intraocular pressure (IOP) must be considered when ophthalmic surgery is to be carried out under general anesthesia. Intravenous anesthetics and volatile agents reduce IOP, with the possible exception of ketamine. Underlying mechanisms include a direct effect on cerebral IOP control centers and indirect effects mediated through the balance between production and drainage of aqueous humor, general circulation and ocular muscle tone. IOP is likely to be elevated during induction and recovery. Currently suggested measures to prevent the increase in IOP associated with laryngoscopic tracheal intubation facilitated by succinylcholine include oral premedication with clonidine, intravenous administration of lidocaine 3 min prior to laryngoscopy, and anesthetic induction with propofol or narcotics. Non depolarizing neuromuscular blocking drugs either do not affect IOP or produce a slight decrease; depolarizing muscle relaxants increase IOP. It remains controversial whether this effect, which is pronounced with succinylcholine, may be reliably abolished by any concomitant medication. The new competitive relaxants atracurium and vecuronium provide stable conditions with respect to IOP and systemic circulation, combined with a rapid onset and intermediate duration of action. PMID- 3052166 TI - [Anesthesia and analgesia in opiate dependence]. AB - This review is devoted to problems that can arise in analgesia and anesthesia in opiate addicts. The following factors are especially important: psychic disorders, heart, lung, liver and kidney disease, AIDS, and tolerance to opiates. Withdrawal of opiates during the perioperative period has a detrimental effect, and the addicted patient should therefore receive generous premedication. Practical experience in the United States of America has shown that the prophylactic administration of methadone is beneficial and prevents withdrawal symptoms. General anesthesia and regional anesthesia are both possible. It is possible to give opiates during general anesthesia achieved with preparations given by inhalation (isoflurane), but it must be borne in mind that high doses are necessary because of these patients' tolerance levels. When regional anesthesia is used for surgery or for postoperative anesthesia, it is important to observe the neurological contraindications. High doses of opiates can be administered (s.c., i.m., p.o., continuous infusion) for postoperative analgesia in opiate addicts with no fear of respiratory depression, with further analgesic treatment as and when necessary (antidepressants, anti-inflammatory agents, neuroleptics, electroacupuncture). Attention is drawn to clonidine as a particularly good drug for the treatment of opiate withdrawal. Problems encountered with abstinent drug addicts are discussed. PMID- 3052167 TI - [Myocardial metabolism as affected by propofol in geriatric patients. A comparison with etomidate]. AB - This study was designed to compare the effects of propofol and etomidate on myocardial metabolism in elderly patients without clinical manifestations of heart failure or coronary artery disease. Twenty geriatric patients (age 65-82 years) scheduled to undergo elective major upper-abdominal surgery were studied and randomly allocated to two equal groups (propofol and etomidate). All patients were premedicated with piritramide, 7.5 mg, and promethazine, 25 mg, intramuscularly 1 h before arrival in the anesthesia room. Ten patients received propofol (1.5 mg/kg) for induction of anesthesia, followed by 10-min infusion of an induction dose; thereafter, anesthesia was maintained with a continuous infusion of 0.1 mg/kg per min. Ten patients received etomidate, 18 mg, for induction, followed by 2.4 mg/min for maintenance. Vecuronium was used for neuromuscular blockade. Cardiovascular dynamics were recorded while the patients were awake, 1-2 min after induction during apnoea, and 1, 5 and 30 min after tracheal intubation without surgical stimulation. Coronary blood flow (argon wash in technique with sampling of blood from the coronary sinus), myocardial oxygen consumption and myocardial uptake of glucose, free fatty acids and lactate were determined in the awake state and 5 and 30 min after intubation. Arterial plasma concentrations of propofol (high-pressure liquid chromatography with fluorescence detection) and etomidate (gas chromatography) were measured every 5 min throughout the investigation period, which lasted 45 min. Overall mean plasma concentrations of propofol were 3.69 +/- 0.16 micrograms/ml and of etomidate 1.1 +/- 0.16 microgram/ml.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3052168 TI - [Induction of anesthesia using propofol in comparison with etomidate]. AB - Etomidate and propofol were compared for induction of anesthesia in a controlled study, including 24 male patients (ASA groups I and II). Following oral premedication with lormetazepam, the patients received propofol (2.5 mg kg-1; n = 12) or etomidate (0.3 mg kg-1; n = 12) over 60 s. For statistical analysis of the cardiovascular data (blood pressure and heart rate) four blocks were set up: A, baseline value including atropine dosage; B, value after induction of anesthesia; C, value after administration of halothane and vecuronium before intubation; D, value after intubation. The blood pressure fell slightly on administration of propofol while the heart rate remained nearly unchanged. Etomidate was associated with unacceptably high increases in blood pressure and heart rate. Myoclonia occurred in seven patients after etomidate and in two patients after propofol. A smoother mask ventilation was rated as a further advantage of propofol. Because of the unfavorable cardiovascular profile, the occurrence of myoclonia and poor mask ventilation, etomidate proved to be unsuitable for induction of anesthesia unless supplemented by an opioid and/or benzodiazepine. The high incidence of pain upon injection was considered to be a disadvantage of propofol. PMID- 3052169 TI - [Patient-controlled analgesia. A technical toy or a contribution to the treatment of pain?]. AB - PCA (patient-controlled analgesia) was used to treat postoperative pain after general surgery and gynecological operations in a total of 82 patients. In a prospective randomized study, 20 of these patients received pentazocine and 20 were treated with Fentanyl. The bolus quantity for pentazocine was 15 mg in 5 ml NaCl, and that for Fentanyl 0.05 mg in 5 ml NaCl. A maximum of 3 boluses was allowed within 1 h; the refractory period was 5 min. Both drugs were equally suited for the treatment of pain. With pentazocine, an average of 144 micrograms kg-1 min-1 was administered during the first 16 h after the operation; with Fentanyl, the quantity taken was 0.78 microgram kg-1 min-1. The inter- and intraindividual variance in the consumption of analgesics described by other authors was confirmed. The amount of analgesics required ranged between 0.05 and 1.95 mg for Fentanyl and between 15 and 435 mg for pentazocine in a period of 16 h. Three patients did not request an analgesic at all. The average consumption of analgesics constantly decreased in the first few postoperative hours, from 0.28 mg every 4 h after the operation to 0.18 mg every 4 h 16 h later (Fentanyl) and from 55 mg every 4 h after the operation to 31.5 mg every 4 h 16 h later (pentazocine). The majority of patients reported very positive experience with PCA. There were few side effects. Problems arose from the negative attitude of other doctors and the nursing staff, and from some misunderstandings.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3052170 TI - Postnatal development of the stomach in the Japanese field vole, Microtus montebelli. PMID- 3052171 TI - [Arterial vascularization of the intestine of the muskrat (Ondatra zibethicus)]. PMID- 3052172 TI - The neocortex of the dog. 1. A classical cytoarchitectonic map. PMID- 3052173 TI - Changes in surface area and number of Leydig cells in relation to the 6 stages of the cycle of the human seminiferous epithelium. AB - In order to evaluate the occurrence of a Leydig cell cycle related to the cycle of the seminiferous epithelium in man, the numbers of peritubular Leydig cells and surface area of these cells along 1 mm of tubular basement membrane at each stage of the cycle were calculated on histological sections of young adult testes. The Leydig cells that were located separated from the tubules (perivascular Leydig cells) were also classified according to the stage of the cycle shown by the nearest seminiferous tubule; the surface area and number of these cells were also calculated. The total surface area and numbers of Leydig cells (peritubular plus perivascular) along 1 mm of tubular basement membrane did not change during the cycle of the seminiferous epithelium. Both the surface area and the numbers of peritubular Leydig cells were greater in stages I and II of the cycle, when spermatozoa are released; they decreased in stages III and IV and increased again in stages V and VI, whereas the contrary occurred in perivascular Leydig cells. The average surface area of each Leydig cell type remained constant throughout the stages of the cycle. PMID- 3052175 TI - [History of Andrology]. PMID- 3052174 TI - Early ontogeny of serotonin-immunoreactivity in the sheep brain. An immunohistochemical study. AB - Using immunohistochemistry with specific antiserotonin anti-sera, the ontogeny of serotonergic neurons was studied in the foetal sheep brain. Serotonergic immunoreactive perikarya first appeared rostrally on day 25 of pregnancy, in the medio-ventral part of the mesencephalic flexure, and caudally, on day 28, in the medio-ventral part of the cervical flexure. The development of this system is very rapid, because on day 40 of gestation, all serotonergic nuclei present in the adult were visible. Compared with other species such as rodents or primates, serotonin appears early in the sheep nervous system, and the development of the serotonergic system is even more rapid. Serotonergic immunoreactivity was seen in some cell bodies in the growing adenohypophysis between days 40 and 50. This phenomenon has not been observed in other species. Because serotonin appears very early and is present in growing areas of the nervous system, it could play a trophic role in the development and maturation of the sheep central nervous system, as has been described previously in other species. PMID- 3052176 TI - ASA Award: Francis F. Foldes. American Society of Anesthesiology. PMID- 3052177 TI - Treatment of non-cardiogenic pulmonary edema following cardiopulmonary bypass with veno-venous extracorporeal membrane oxygenation. PMID- 3052178 TI - [A review of the mammalian Mallophaga of Europe]. AB - The paper reviews the present knowledge of the Mallophaga of European mammals. 50 species and subspecies have been listed in a checklist and a host-parasite list. 32 of them are still found in Europe. There have been no records so far of 21 hosts listed as hypothetical mallophagian hosts. Eight extinct or exterminated forms of mammals in Europe are listed as hypothetical hosts of chewing louse. PMID- 3052179 TI - [Validation of ultrasonic flowmetry and occlusion plethysmography in venous thromboses]. PMID- 3052180 TI - [Aneurysm of the abdominal aorta: results of surgical treatment]. PMID- 3052181 TI - [The effect of dihydroergotamine on the venous system in patients with essential varices of the lower limbs]. PMID- 3052182 TI - The carpal tunnel syndrome--a disease underlying Raynaud's phenomenon? AB - Forty patients with clinically and electromyographically proven carpal tunnel syndrome were examined for the frequency of concomitantly occurring Raynaud's phenomenon. The angiologic work-up was based on a detailed vascular history and examination, Doppler sonography to determine systolic digital artery pressures before and after exposure to the cold (10 degrees C for one minute), and thermoplate tests for recording heat convection of the hands at normal room temperature and measuring rewarming time after standardized cold exposure. Only 10% of the patients were found to have manifest Raynaud's phenomenon. The incidence was thus the same as in the general population. In addition, Raynaud's phenomenon and the carpal tunnel syndrome were poorly correlated with respect to their preliminary symptom appearances. Consequently, the carpal tunnel syndrome can no longer be regarded as a disease underlying Raynaud's phenomenon. PMID- 3052183 TI - Assessment of captopril and nicardipine effects on chronic occlusive arterial disease of the lower extremity using Doppler ultrasound. AB - The effects of two potent vasodilating drugs, captopril (C) (25 mg tid), nicardipine (N) (20 mg tid), and placebo (P) were evaluated and compared, in 10 men (mean age of sixty-five years) with intermittent claudication from moderate to severe multilevel chronic occlusive arterial disease (COAD) of the lower extremity, by use of the Doppler ultrasonic method, at rest and after Carter's exercise test. All the examined subjects were normotensive, without diabetes or cardiopathy; all have been smokers. The eight-week total protocol consisted of an initial two-week placebo run-in period followed by two active drug phases and a two-week placebo phase, according to a double-blind, randomized, crossover design. At the end of each two-week period, ankle-arm index (AAI) and, following exercise, onset of lower extremity discomfort time (ODT), duration of exercise (ET), decrease of ankle systolic pressure after test (APD), and recovery time (RT) were determined. Moreover, at rest, just after exercise, and after recovery, simultaneous common femoral artery velocity waves were recorded and analyzed by a quantitative approach to detect the peripheral vasomotor adjustments. None of the patients required the withdrawal of the active treatments. Compared with P, C significantly reduced APD and RT, and N reduced RT and AAI; furthermore N caused a significant decrease in ODT, whereas C showed a trend, although not statistically significant, to increase ODT. Neither active therapy modified ET. These results suggest that C and N have different short-term effects on peripheral circulation in COAD. During exercise, C induces hemodynamic improvement in the ischemic lower extremity probably by inhibition of the sympathetic system and consequent reduction in collateral vessel vasoconstriction. PMID- 3052184 TI - Disturbances of cutaneous microcirculatory autoregulation in arterial occlusive disease. AB - Skin blood flux at the dorsum of the foot was measured during local heating--by use of laser Doppler flowmetry--in 6 healthy subjects, 6 claudicants, and 6 patients with rest pain. Significantly different flux increases were noted among the three groups during the first six minutes of heating. Healthy subjects showed an initial rapid increase, followed by an abrupt reduction influx through a probable myogenic autoregulatory microcirculatory control mechanism. In patients the flux increase was slow and continuous, well correlated with the distal perfusion pressure; autoregulatory control was diminished or absent. With less than 100 mmHg perfusion pressure the microvessels are probably constantly dilated through a reduced sympathetic vasoconstriction tone, and further autoregulatory mechanisms are impaired. Measuring of functional behavior of cutaneous microcirculation under low arterial inflow pressure could represent a method for sensitive diagnosis of arterial insufficiency. PMID- 3052185 TI - Value and limits of "critical auscultation" of neck bruits. AB - Within a group of 2,000 patients evaluated, most of them with symptoms of cerebrovascular insufficiency, 441 had a monolateral or bilateral cervical bruit. The 627 sides with an audible bruit were divided into main groups (A) symptomatic (TIA and/or stroke homolateral to the bruit), (B) possibly symptomatic (non-side related symptoms), (C) asymptomatic (C1, in totally asymptomatic patients; C2, in patients with symptoms dependent on the hemisphere contralateral to the cervical bruit). Each patient was studied by means of clinical (history, blood pressure in both upper limbs, phonoendoscopic auscultation at various levels) and noninvasive instrumental examinations (CW Doppler spectrum analysis). An apparently primitive cervical bruit corresponded to a lesion of the carotid bifurcation in 61% of the cases (positive predictive value) whereas a normal bifurcation was detected in 70% of the cases in which the cervical bruit was considered as secondary (negative predictive value); the diagnostic accuracy of the "critical auscultation" has a value therefore of 63%, with a sensitivity of 84% and a specificity of 40%. The results obtained in the different groups of patients (symptomatic or asymptomatic) were not significantly different (chi-square). Even though maintaining the value of a cervical bruit as a sign of carotid stenosis or occlusion and consequently confirming the importance of neck auscultation, the authors conclude that the critical auscultation as commonly performed is not capable of excluding the presence of a carotid lesion with sufficient reliability, even in totally asymptomatic patients. PMID- 3052186 TI - A new clinical sign for the diagnosis of allergic conjunctivitis: a brief communication. AB - A clinical sign of the lower eyelid to be referred to as the "Mag Sign" or "Crescent Sign" is described, which when present is highly suggestive for conjunctivitides of allergic origin mostly due to aeroallergens (IgE-mediated type I allergic reaction). It is hoped that this sign will facilitate the diagnosis and treatment of allergic conjunctivitis. PMID- 3052187 TI - Beneficial effect of prostaglandin E1 in three cases of lupus nephritis with nephrotic syndrome. AB - Prostaglandin E1 was intravenously injected daily for 4 weeks to three patients with lupus nephritis accompanied by severe nephrotic syndrome. Histologic examination revealed diffuse proliferative or membranous glomerulonephritis. Proteinuria decreased markedly 1 year after the treatment with improvement in hypoproteinemia in all three patients. Five years later, the conditions of three cases ameliorated further. PMID- 3052189 TI - Importance of food challenge procedures. PMID- 3052188 TI - Comparative tolerability of two formulations of Rhinalar (flunisolide) nasal spray in patients with seasonal allergic rhinitis. AB - This double-blind, randomized, crossover study compared the incidence of nasal burning and stinging, as well as overall tolerability of the currently marketed formulation of Rhinalar (original formulation) to a new formulation of Rhinalar containing less propylene glycol. In addition, patient and investigator subjective evaluations were used to compare the effectiveness of the test medications in controlling the nasal symptoms of seasonal allergic rhinitis. A total of 122 patients were enrolled in this 4-week trial. Each patient received one formulation of Rhinalar for 2 weeks and then crossed over to receive the alternate formulation for an additional 2 weeks. Eighteen patients withdrew from the trial prematurely. Ten patients were lost to follow-up and eight withdrew due to side effects and/or inadequate therapeutic response. Statistical comparisons of patient evaluations of nasal burning and stinging with the two formulations of Rhinalar showed a very significant difference in terms of severity (P less than .001), duration (P less than .001), and tolerability (P = .006) in favour of the new formulation. A reduction in severity of throat irritation with the new formulation was also shown to be statistically significant (P = .006). Nausea, headache, and other side effects including watery eyes, taste perversion, and runny nose were seldom reported with either test medication. Both formulations were shown to be equally effective in relieving the nasal symptoms of seasonal allergic rhinitis. The considerable reduction in nasal burning and stinging and throat irritation with the new formulation of Rhinalar was shown to enhance patient acceptability and may lead to better compliance. PMID- 3052190 TI - [Biological diagnosis of hereditary metabolic diseases. From selective screening to the mutant-cell bank]. AB - The experience of a specialized laboratory for the biological diagnosis of inborn errors of metabolism in selected pediatrics patients is reported. The strategy starts with a wide testing of blood and urine, as many inborn errors of metabolism can be detected through testing of blood and urine for increased concentration of specific metabolites known to be associated with the genetic defect. Then enzymatic or DNA studies are performed to confirm the diagnosis. The mutant cells mostly fibroblasts are stored in a cell bank and available for other research. PMID- 3052191 TI - Hereditary angioedema. AB - Although the condition is rare, patients with hereditary angioedema often present because of abdominal pain or airway compromise. A 27-year-old woman presented to the emergency department in acute abdominal distress. Identification of the disease in this patient allowed for proper management and avoidance of invasive procedures. Pathophysiology, clinical manifestations, diagnosis, and therapy of hereditary angioedema are discussed. PMID- 3052192 TI - Effects of frequency and airway pressure on gas exchange during interrupted high frequency, positive-pressure ventilation in ponies. AB - Cardiovascular effects and pulmonary gas exchange were compared during conventional mechanical ventilation (CMV) and interrupted high-frequency, positive-pressure ventilation (IHFPPV) in 6 anesthetized ponies in dorsal recumbency. When the peak airway pressure (Paw) was held constant at control values attained during CMV (18 to 20 cm of H2O), and the ventilator frequency of IHFPPV was varied over the range, 2.5 to 12.5 Hz, significant (P less than 0.05) changes from control values were observed only in the ratio of dead-space volume to tidal volume (VD/VT) and in the respiratory minute volume (VE). The mean (+/- SEM) carbon dioxide excretion (VCO2) was 2.12 +/- 0.1 ml/kg/min during IHFPPV. Dead-space ventilation ranged from 40 to 73.7% of total ventilation and increased directly with increasing frequency. The VE also increased, from 89 ml/kg/min at a ventilatory frequency of 2.5 Hz to 145 ml/kg/min at a frequency of 12.5 Hz. Maintaining the frequency of IHFPPV constant at 12.5 Hz and increasing the Paw over the range of 5 to 30 cm of H2O caused significant (P less than 0.05) changes in arterial partial pressure of O2 (PaO2), VCO2, pulmonary shunt fraction (QS/QT), VE, arterial-alveolar differences in oxygen tension (AaDO2), VD/VT, and cardiac output, compared with CMV. The PaO2 and the VCO2 increased linearly with increasing Paw. With increasing Paw, VD/VT decreased directly with increasing Paw from 98 to 69.3%. Gas exchange at a Paw of 15 cm of H2O during IHFPPV was equivalent to conditions at Paw of 20 cm of H2O during CMV. At a higher Paw during IHFPPV, improvements over control values were observed in gas exchange. PMID- 3052193 TI - Differences in diagnostic test results and hematologic data between aged and young horses. AB - Hematologic data and results of diagnostic tests were compared between aged (greater than or equal to 20 years old) and young (less than or equal to 5 years old) horses to identify hematologic and metabolic changes associated with aging. Initial data were obtained from 8 aged and 6 young mares (group 1). Similar data were collected from a second group of aged (3 mares and 3 geldings) and young (1 mare and 5 geldings) horses (group 2). Dexamethasone suppression tests (DST) and necropsies were performed on 6 additional mares and mare 8 from group 1 (group 3). Complete blood counts and serum biochemical profiles were compared between young and aged horses of groups 1 and 2. Mean corpuscular volume was higher (P less than 0.05) in aged horses. Oral glucose tolerance and insulin response to orally administered glucose were measured in 13 aged horses (groups 1 and 2) and 6 young mares of group 1. In group 1, plasma ascorbic acid values were lower (P less than 0.05) in aged horses than in young horses maintained under the same conditions and feeding regimens. An apparent age-related hyperinsulinemic response to orally administered glucose identified in group-1 mares was probably a result of a high occurrence of subclinical hypophyseal and/or thyroid adenomas. Of 13 aged horses necropsied (groups 2 and 3), 10 had hypophyseal and/or thyroid adenomas that, in group 2, were consistently associated (P less than 0.05) with hyperinsulinemic responses to orally administered glucose. All horses in groups 2 and 3 were given a 24-hour DST.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3052194 TI - Specific serodiagnosis of canine leishmaniasis by indirect immunofluorescence, indirect hemagglutination, and counterimmunoelectrophoresis. AB - The increased prevalence of leishmaniasis in dogs in Mediterranean countries has necessitated use of a reliable specific method of immunodiagnosis. Research was carried out on 88 dogs, among which were 26 with severe clinical leishmaniasis (group A), 15 with mild signs of disease (group B), and 11 without signs of disease (group C); in addition, 15 were healthy (group D) and 21 were affected with various other diseases (group E). Dogs of groups D and E were used as controls. Infection was confirmed by cultural isolation of the parasite. Serotests that were used included the indirect immune fluorescent (IFAT) and indirect hemagglutination (IHAT) tests and counterimmunoelectrophoresis (CIEP); using these tests, specific antibodies against Leishmania infantum were detected in all 88 dogs. Sensitivity of the IFAT was 100% in dogs of groups A, B, and C; for the IHAT, it was 65.38%, 60%, and 54.5%, respectively, and for CIEP, it was 96.1%, 80%, and 72.7%, respectively. For dogs of groups D and E, specificity of the IFAT and IHAT was 100%, and for CIEP, it was 100% for group-D dogs and 90.5 for group-E dogs. Indirect immunofluorescence appeared to be the most specific and sensitive of the 3 tests. PMID- 3052196 TI - The meanings of personality predicates. PMID- 3052195 TI - Effect of flunixin meglumine on Escherichia coli heat-stable enterotoxin-induced diarrhea in calves. AB - In a study to evaluate the effect of flunixin meglumine on secretory diarrhea, 11 calves were assigned to 3 groups: group 1 (n = 3) served as controls, group-2 calves (n = 4) were given 2.2 mg of flunixin meglumine/kg, IM at 7 AM and 3 PM, and group-3 calves (n = 4) were given 2.2 mg of flunixin meglumine/kg, IM at 7 AM, 11 AM, and 3 PM. All calves were given approximately 200 micrograms of heat stable Escherichia coli enterotoxin (STa) orally at 8 AM. Mean cumulative fecal output for groups 1, 2, and 3 was 1,331.0 +/- 317.2 g, 1,544.3 +/- 154.4 g, and 785.5 +/- 276.5 g, respectively. There was a significant (P less than 0.05) reduction in mean fecal output in group-3 calves, compared with that in groups 1 and 2. Calves in group 2 tended to have a delay, but not a reduction, in their fecal output. At 12 hours, hemoconcentration was significantly (P less than 0.05) greater in group-1 calves than in group-2 or group-3 calves. PMID- 3052197 TI - The challenge of double jeopardy. Toward a mental health agenda for aging women. PMID- 3052198 TI - The relevance and irrelevance of psychological research. The example of prison crowding. PMID- 3052199 TI - The nature-nurture debate, aptitudes, and group differences. PMID- 3052200 TI - Appendicitis. The diagnostic challenge continues. AB - The typical clinical manifestations of appendicitis are well described, but atypical presentations occur frequently. Uncertainty regarding the diagnosis may cause long delays before appropriate treatment is rendered. Several patients in whom the diagnosis was obscure are presented. Barium enema, ultrasonography, computerized tomography, and laparoscopy each may be helpful in diagnosis, but these studies should be used selectively and negative results can be misleading. It is wise to consider other causes of abdominal pain in patients suspected of having appendicitis, so that unnecessary operation can be avoided. It is more important to suspect appendicitis in patients who appear to have nonsurgical conditions of the abdomen, so that the incidence of perforation can be minimized. PMID- 3052201 TI - Complete cerebral angiography in the evaluation of patients with cerebrovascular insufficiency. AB - Three hundred and twenty-eight consecutive patients presenting with symptoms of cerebrovascular insufficiency were evaluated with complete cerebral angiography (CCA) under a prospective protocol evaluating the necessity and the utility of CCA over diagnostic modalities confined in their scope to the carotid bifurcation. Procedure mortality was 0.3 per cent (one fatal stroke), major morbidity 0.6 per cent (one non-fatal stroke and one stroke with resolution), and minor morbidity 2.1 percent. Eighteen per cent (60/328) of the CCAs demonstrated highly significant intracerebral pathology that materially affected choice of therapy. CCA was as valuable in evaluating patients with transient ischemic attack, 22 per cent (27/121), as prior stroke, 23 per cent (22/98). CCA is highly useful for the accurate evaluation of associated disease outside the carotid bifurcation, and is therefore essential for accurate delineation of intracerebral anatomy, pathology, and flow patterns. It may be performed with reasonable patient risk, provides information not available from, or of better quality than, real time ultrasound or intravenous digital subtraction angiography, and therefore is essential for the proper selection of procedures, if any is necessary, and the proper order of those procedures. PMID- 3052202 TI - [Insulin receptors in intrauterine growth retardation]. AB - This report analyses erythrocyte insulin binding and specific erythrocyte insulin receptors in the neonatal period. Authors have studied thirty normal newborns who had an adequate weight for their gestational age and another fifteen who were asymmetric intrauterine retarded growth. Cord blood samples were collected and insulin binding, insulin receptors, blood glucose concentration, serum immunoreactive insulin, serum C-peptide concentration and molar ratio of C peptide to insulin were determined. Insulin and C-peptide sera concentrations suggest that basal insulin secretion in small for data newborns is similar to term infants. Significantly minor affinity constant (p = 0.05) and significantly major dissociation constant (p = 0.05) in small for date infants versus newborns adequately weighted for their gestational age, justify the increased sensitivity to insulin as it happens in other states of chronic undernutrition. In small for date infants, number of sites per cell correlates negatively (r = -0.57, p less than 0.05) with birth weight. Up regulation mechanism that modulates receptors concentration could explain this phenomenon. PMID- 3052203 TI - [Fasciitis with eosinophilia: Shulman syndrome. Report of a case and review of the literature]. AB - A twelve year-old boy who developed, after a period of strenuous physical work, an illness characterized by thickened skin over his right thigh and hemiabdomen, flexion contractures in right wrist and elbow and in right metacarpophalangeal joints without Raynaud's phenomenon or other visceral symptoms is presented. Pertinent laboratory studies showed hypereosinophilia and hypergammaglobulinemia. Deep-fascia biopsy showed typical findings of fasciitis with eosinophilia. Prednisone therapy resulted in sustained improvement. Periarticular osteopenia of the right hand and mast cell infiltration in fascial biopsy are remarkable features. Authors stress striking differences between fasciitis with eosinophilia or Shulman's syndrome and scleroderma. Clinical picture, laboratory changes, typical histology and a usually rapid response to corticosteroids, as well as some autoimmune diseases possibly associated, suggest a different disease and an immunological pathogenesis. From literature review authors conclude that cases of Shulman's syndrome reported in children are very limited and that this disease should by ruled out in every child with thickened skin changes and articular flexion contractures. PMID- 3052204 TI - [Mollusks of parasitologic and veterinary interest in Italy]. PMID- 3052205 TI - [Erickson's hypnosis]. AB - Milton H. Erickson (1901-1980) renovated the study and practice of therapeutic hypnosis. The author first presents a synthetic overview of Erickson's original work and its spread. He then illustrates this with excerpts from observations of six of his own patients which correspond to the progressive integration of an ericksonian approach into a classical psychiatric practice in a general hospital setting. PMID- 3052206 TI - [The psychological preoccupation of Sir James Paget, and contribution to medico surgical psychology]. PMID- 3052207 TI - Tissue destruction resulting from the interaction of cytotoxic T cells and their targets. AB - In vitro and in vivo-generated cytotoxic T lymphocytes (CTL) specific for major and minor histocompatibility antigens evoked antigen-specific full-thickness skin necrosis when injected intradermally into allogeneic mice in a variety of strain combinations. In addition, CTL-target-cell mixtures injected intradermally into hosts syngeneic to the CTL also evoked destruction of host tissue. These "innocent bystander" reactions were evoked with alloreactive CTL as well as with CTL directed against hapten (TNP)-modified and virus (influenza A)-infected target cells. Unlike the direct reactions, the bystander reactions in histocompatibility-antigen systems occurred in spite of H-2 incompatibility of the CTL, admixed target cells, and the hosts. One explanation for these results, currently under investigation, is that some bystander reactions may occur without MHC restriction. In aggregate, our findings indicate that nonspecific as well as antigen-specific reactions initiated by CTL-target-cell interactions may contribute to tissue destruction in allograft rejection, in severe forms of delayed-type hypersensitivity, and in certain viral infections. PMID- 3052209 TI - Recovery from acute virus infection. Role of cytotoxic T lymphocytes in the elimination of lymphocytic choriomeningitis virus from spleens of mice. PMID- 3052208 TI - Functional analysis of CD2, CD4, and CD8 in T-cell activation. PMID- 3052210 TI - Mechanism of natural killer cell-mediated cytotoxicity. PMID- 3052211 TI - A molecular-genetic analysis of cytotoxic T lymphocyte function. AB - Two genes that are specifically expressed in T cells with cytolytic activity were isolated from a CTL cDNA library by differential screening. Both appear to encode serine proteases, thus suggesting a cascade mechanism, similar to complement, in activated CTL. Both CTL-specific proteases have a number of unusual structural features that suggest that they will have novel substrate specificities. One of the proteins (CCPI) has been oriented to the granules found in the cytoplasm of CTL. Taken together, these data strongly suggest that these molecules play an important role in target-cell lysis by CTL. Furthermore, we believe that the detailed molecular knowledge being accumulated through these studies may lead to the development of innovative forms of immunotherapy. PMID- 3052212 TI - A T cell- and natural killer cell-specific, trypsin-like serine protease. Implications of a cytolytic cascade. AB - A new trypsin-like serine protease was cloned from both a murine cytotoxic T lymphocyte and a human PHA-stimulated peripheral blood lymphocyte cDNA library. In both the mouse and human system, this transcript had a T cell- and NK-specific distribution, being detected in cytotoxic T lymphocytes (CTL), some T-helper clones, and NK, but not in a variety of normal tissues. T-cell activation with Con A plus IL-2 induced mouse spleen cells to express this gene with kinetics correlating with the acquisition of cytolytic capacity. Both the mouse and human nucleotide sequences of this gene encoded an amino acid sequence with 25-40% identity to members of the serine protease family. The active-site "charge-relay" residues (His-57, Asp-102, and Ser-195 of the chymotrypsin numbering system) are conserved, as well as the trypsin-specific Asp (position 189 in trypsin). We reviewed the evidence of this serine protease's role in lymphocyte lysis and proposed a "lytic cascade." We discussed the biological and clinical implications of a cascade, proposing these enzymes as markers for cytolytic cells and as targets for rational drug therapy. Genetic and acquired deficits in the lethal hit-delivery system are considered as a basis for approaching some immunodeficiency states, including severe EBV infections, T-gamma leukemias, and T8+ lymphocytosis syndromes. PMID- 3052213 TI - Role of soluble cytotoxic factors in lymphokine-activated killer cell (LAK) mediated cytotoxicity. PMID- 3052214 TI - Direct and indirect modes of action of cyclosporine on cytotoxic T lymphocytes. PMID- 3052215 TI - Phenotypic identification of memory cytolytic T lymphocytes in a subset of Lyt-2+ cells. PMID- 3052216 TI - Expression of T-cell receptors by functionally distinct subsets of immature adult thymocytes. PMID- 3052217 TI - Presumed intraocular nocardiosis in a cardiac-transplant patient. AB - A 44-year-old cardiac-transplant patient receiving immunosuppressive therapy developed a chorioretinal mass OS. The lesion was consistent with an ocular nocardial infection. The patient also developed a testicular abscess that was shown to contain N. asteroides after orchiectomy. Despite appropriate systemic treatment, the patient developed extensive subretinal scarring OS. This case report is the first to describe an opportunistic ocular infection with N. asteroides after cardiac transplantation. PMID- 3052218 TI - [Artificial filiation. Medico-social, technical and legal prospectives of artificial procreation]. PMID- 3052219 TI - [Doppler velocimetry: differential diagnosis of small fetus and intrauterine growth retardation]. PMID- 3052220 TI - Congenital stenosis of the lower esophagus associated with leiomyoma and leiomyosarcoma of the gastrointestinal tract. AB - The purpose of this paper is to report our analysis of four generations of a family with congenital strictures of the lower esophagus associated with leiomyoma of the gastrointestinal tract. Two members of the family died of sarcomatous degeneration of the leiomyoma and one is still alive 2 years after resection of the malignancy. Three family members had surgical repair of the congenital stricture. Surgical repair was unnecessary in one, for whom repeated dilatations alleviated the symptoms. Since there were no direct female descendants in this family, it is unknown whether this hereditary disease is carried by a sex-linked dominant gene. There are no reports in the literature of a familial tendency to congenital strictures of the esophagus, nor are there any reports of strictures in association with gastrointestinal leiomyoma and the later development of sarcomatous degeneration. The literature is reviewed, syndromes of leiomyoma of the esophagus and gastrointestinal tract are detailed, the particulars of the family tree and the patients are described, and the study is summarized. PMID- 3052221 TI - Dilation of esophageal strictures: comparative morbidity of antegrade and retrograde methods. AB - A total of 687 dilations of esophageal strictures were performed on 59 patients in the operating room over a 17-year period. Seventy-nine percent of the strictures were secondary to caustic ingestion and 89% of the dilations were in these patients. Antegrade dilations were performed 389 times and retrograde dilations were performed 298 times. Esophageal perforation occurred seven times with antegrade dilations. There were no perforations with retrograde dilations. The retrograde method using Tucker bougies is the safest and most successful method of dilating severe strictures. PMID- 3052222 TI - Laryngotracheal separation for intractable aspiration. AB - Intractable aspiration may be a life-threatening problem for patients with altered laryngeal function secondary to neurologic disorders or abnormal laryngeal anatomy. Multiple surgical procedures have been devised to deal with this problem. An effective technique involves the creation of a tracheostoma and closure of the larynx at the first or second tracheal ring. Laryngotracheal separation is relatively easy to perform and potentially reversible. Experience with this technique in six patients who required laryngeal separation for intractable aspiration is described. The procedure was successful in preventing aspiration and recurrent pneumonia associated with neurologic dysfunction, unresectable neoplasm, and conservation laryngeal surgery. One patient of one has had a successful reconstruction. PMID- 3052223 TI - Indications for the tracheoesophageal diversion procedure and the laryngotracheal separation procedure. AB - Impaired protective function of the larynx can lead to intractable aspiration, a severe and potentially fatal disorder. If medical therapy fails to prevent intractable aspiration, surgical separation of the upper respiratory tract from the digestive tract is necessary to prevent recurrent contamination of the respiratory system in these patients. Two such surgical procedures are the tracheoesophageal diversion procedure and the laryngotracheal separation procedure. Our approach to patients with intractable aspiration and the indications for the use of these surgical procedures for the prevention of aspiration are discussed. PMID- 3052225 TI - Rib cartilage grafts for the treatment of posterior glottic and subglottic stenosis in children. AB - Posterior glottic and subglottic stenosis from endotracheal intubation in children can be managed endoscopically with varying success. Open surgical treatment offers a better potential for correction with a single procedure in moderate and severe cases. The open method consists of splitting the scar and cricoid cartilage posteriorly to the level of the interarytenoid muscle, then stenting the incised cricoid open with a rib cartilage graft. Use of this method is described, and results in 12 cases are reported. Decannulation was achieved in ten patients. In all patients who were decannulated, good exercise tolerance, freedom from aspiration, and an adequate voice quality were achieved. PMID- 3052224 TI - Upper aerodigestive tract manifestations of cicatricial pemphigoid. AB - Cicatricial pemphigoid is a chronic mucosal blistering disorder with a predilection for subsequent scar formation. Many physicians may be unaware of the various presentations and sequelae of this uncommon disease. This report of the largest series to date focuses on the upper aerodigestive tract manifestations of this disease. During the years 1975 to 1985, 142 patients with cicatricial pemphigoid were seen at the Mayo Clinic. There were 93 women and 49 men; the age range was 21 to 92 years. Mucosal lesions occurred most often in the mucous membranes of the oral cavity and conjunctiva. Involvement of the pharynx, larynx, and esophagus was less common. Stenosis of the nasopharynx or larynx necessitated surgical repair in several persons and caused obstructive sleep apnea in two. The otolaryngologist can make an important contribution to the early recognition, diagnosis, and management of the complications of cicatricial pemphigoid. PMID- 3052226 TI - Bronchoesophageal manifestations of acquired immunodeficiency syndrome. AB - Among the more common manifestations of acquired immunodeficiency syndrome (AIDS) are tumors and infections that occur in regions treated by the bronchoesophagologist. In reviewing our institutional experience in the diagnosis and treatment of 396 patients with AIDS in 1987, we have noted that 226 (57%) had some form of pneumonia and 133 (34%) had candidiasis. In this communication we discuss the various types of bronchopulmonary, oropharyngeal, and esophageal infections that have been reported among AIDS patients. We also review the universal precautions and specific guidelines recommended for safeguarding the bronchoesophagologist and other health care workers who treat these patients. PMID- 3052227 TI - Near-fatal complication of tracheal T-tube use. AB - Stenosis of the larynx and trachea is an unfortunate sequel to many thermal injuries. Numerous surgical techniques have been developed for correction of such problems, many involving use of a tracheal T-tube. We report a serious complication attributed to the use of such a tube. Factors contributing to this complication are analyzed and methods for avoiding similar near-catastrophes discussed. PMID- 3052228 TI - Kirschner wire migration through the jugular foramen. PMID- 3052229 TI - Cervical ranulas. AB - Cervical ranulas, known also as plunging or burrowing ranulas, are an outcome of extravasated sublingual gland mucin that has gained access to the soft tissues of the neck. These pseudocystic lesions may be localized or extensive, and they require surgical excision of the sublingual gland for effective management. PMID- 3052230 TI - [Current status of bronchopulmonary dysplasia]. PMID- 3052231 TI - [Neonatal herpes infection]. PMID- 3052232 TI - [Algodystrophy in children]. PMID- 3052233 TI - Immediate skin grafting on microvascular free tissue transfers. AB - Clinical reports of pedicled muscle flaps and microvascular tissue transfers include two distinctly different strategies for skin grafting. Some authors describe immediate grafting, whereas others recommend a three- to 5-day interval between flap transfer and grafting. No systematic analysis of either strategy has been reported. We reviewed 51 consecutive successful free tissue transfers that involved immediate skin grafting. Ninety-six percent of the grafts survived without complication. Bot muscle and fascial flaps supported skin grafts. Skin grafts survived infected recipient sites, pedicle revisions, partial flap necrosis, and secondary operation. We conclude that immediate skin grafting on free tissue transfers is reliable. PMID- 3052234 TI - Combined orthodontic-surgical treatment of alveolar clefts. AB - After more than four years of postoperative follow-up, 224 patients receiving secondary bone grafting of alveolar clefts were examined. Patients were classified into three groups according to age and eruption stage of the cleft side canine tooth at surgery. Each group was then subdivided according to the diagnoses cleft lip alveolar process only, unilateral cleft lip and palate, and bilateral cleft lip and palate. The treatment results were evaluated with respect to bone level in the cleft area, gingival and periodontal condition, orthodontic treatment result, growth of the jaws after surgery, and complications. The best results were achieved when the bone grafting was performed before eruption of the cleft side canine. In half of these patients orthodontic treatment could be finished with a closed dental arch without the need for prosthodontic treatment. The sagittal growth of the jaws was unaffected by the bone grafting, whereas the anterior height of the maxilla was reduced apparently without any clinical significance. PMID- 3052236 TI - Reconstruction of ear helix: use of self-tubing pedicle flap. AB - The tube pedicle flap is useful in reconstruction of an absent ear helix. However, immediate tubing of the flap may result in vascular compromise. By elevating a bipedicle flap in the postauricular region and shielding it with a silicone sheet, it is possible to allow the flap to assume a tubular shape while preventing adherence to the underlying suture line. Once mature, the flap maintains both a cylindrical shape and a sufficiently thin external diameter to provide adequate restoration of the ear helix. PMID- 3052235 TI - Congenital ear deformity: reconstruction using composite graft. AB - We present a congenital deformity of the right ear, characterized by absence of the central portion of the helix and antihelix. The deformity was reconstructed by advancing retroauricular skin and closing the resultant defect with a composite graft from the posterior aspect of the left ear. This technique was performed in a single stage and required no incisions on the anterior surface of either ear. Postoperative appearance of the right ear was satisfactory, and donor deformity was minimal. PMID- 3052237 TI - External longitudinal splitting of the biceps brachii muscle for coverage of repaired brachial vessels: an anatomical study and clinical application. AB - The external longitudinal splitting of the biceps brachii muscle offers a convenient and safe method for covering exposed vessels in the medial aspect of the arm in cases with loss of soft tissue. This anatomical study reveals rather variable and inconsistent patterns of blood supply to the biceps brachii muscle. The common denominator among the patterns is the fact that the nutrient vessels to the muscle originate in its posterior deep surface and course thereafter in a fan shape upward to the surface of the anterior aspect of the muscle. This microcirculatory pattern permits the external longitudinal splitting of the biceps brachii muscle along the whole anteromedial aspect of the short-headed belly without compromising its blood supply and function. An illustrative case is described. PMID- 3052238 TI - Morphologic alterations of the temporomandibular joint in a patient with Sotos' syndrome: report of a case. PMID- 3052239 TI - Dentinogenic ghost cell tumor presenting as a gingival mass. PMID- 3052240 TI - Bilateral asymmetrical transposition of teeth. Report of a case. PMID- 3052241 TI - Three-rooted mandibular first premolar tooth. PMID- 3052242 TI - Detection of cholinesterase inhibition. The significance of cholinesterase measurements. AB - Human cholinesterase exists in two forms--acetylcholinesterase located in tissue microsomes and red blood cells and serum cholinesterase found in serum or plasma. The two enzymes display marked differences in structure, substrate specificity, biological function, and origin. Contemporary methods employ acylthiocholine as substrate for serum cholinesterase and a second coupled reaction of thiocholine and chromogenic disulfide agents. Clinical applications are primarily centered on subnormal levels of enzyme activity. The decreased activity levels can be caused by inhibitors, reduced biosynthesis, or dysfunctional genetic variants. Changes in enzyme activity should be related to baseline levels because there is wide individual variation as well as methodological variation. Once baseline levels have been established, cholinesterase activity becomes a sensitive indicator of pesticide intoxication and hepatic biosynthetic capacity. A more sophisticated assay, performed in the presence of an inhibitor, is required to detect the atypical genetic variants of serum or plasma cholinesterase. PMID- 3052243 TI - Characterization of neutrophil agglutinins in primary autoimmune neutropenia of early childhood. AB - Neutrophil agglutinins that caused primary autoimmune neutropenia in six young children less than two years of age reported earlier were investigated further for antibody identification and specificity using granulocyte agglutination technique. Initially, the patient sera were tested with parental neutrophils and further confirmed with several donor cells. The sera were tested with autologous neutrophils after partial or complete recovery of the patients from neutropenia. The pattern of reactivity of the patient sera with parental, donor, or autologous neutrophils was compared with that of standard neutrophil typing sera currently available for the antigens NA1, NA2, NB1, NC1, and 9A. The specificity was confirmed by absorbing and retesting the sera with appropriate antigen positive and negative donor neutrophils. Our data revealed anti-NA1 specificity in four and anti-NA2 in two sera. Our observations together with that of McCullough et al suggest that NA1 and NA2 antigens are frequently associated with primary AINI in young children as compared to other neutrophil antigens. PMID- 3052244 TI - Biochemical and genetic identity of alpha-keto acid reductase and cytoplasmic malate dehydrogenase from human erythrocytes. AB - We have recently shown that cytoplasmic malate dehydrogenase (MDH-s) from several non-human species catalyses the reduction of aromatic alpha-keto acids in the presence of NADH (Friedrich et al. 1987), an activity previously attributed to the enzyme aromatic alpha-keto acid reductase (KAR E.C.1.1.1.96). Here we present evidence that this also occurs in humans, and that the previously characterized human KAR is not the product of a genetically distinct locus. Human MDH-s and KAR activities co-migrate after starch gel electrophoresis, and electrophoretic variants of human MDH-s exhibited identical variation for KAR. Both enzymes show almost no electrophoretic variation among human populations of diverse origin. The reduction of aromatic alpha-keto acids is substantially inhibited by malate, the end-product of the MDH reaction. Antibodies raised against purified chicken MDH-s equally inhibited both MDH-s and KAR in chickens and humans. The bulk of the KAR activity in human blood appears to be due to MDH-s, with a minor fraction catalysed by LDH, as is the case in most other species studied. The previous assignment of a gene for KAR to human chromosome 12 in human/Chinese hamster somatic cell hybrids is questioned because interspecific hybrid bands of both MDH s and LDH appear with slightly different mobility approximately midway between the human and hamster controls in somatic cell hybrid studies, and the meaning of this artifact is discussed. The discovery that MDH reacts with intermediate metabolites of phenylalanine and tyrosine has implications in relation to the mechanism by which mental retardation may be produced in phenylketonuria (PKU), and the effect of MDH inhibition on oxidative phosphorylation in the various tyrosinaemias is discussed. PMID- 3052245 TI - Relationship between Ki-67 positive cells, growth rate and histological type of human intracranial tumors. AB - The proliferation rate of 40 intracranial neoplasms (30 gliomas, 1 hemangioblastoma, 3 meningiomas, 1 neurinoma and 5 brain metastases) was investigated using the monoclonal antibody Ki-67. In eleven of the gliomas recurrences could be observed, and two of them recurred for second time. In total the Ki-67 labelling indices of 53 specimens were investigated. The Ki-67 nuclear antigen was demonstrated in frozen sections by application of the appropriate monoclonal antibodies according to a modified alkaline phosphatase-antialkaline phosphatase (APAAP) technique. The proliferation rate was evaluated by cell count calculation of the staining index. Ki-67-labelled glioma cells varied from 0.2 percent in one meningioma (WHO-grade I) to 9.1 percent in one glioblastoma. In ten glioma recurrences, higher Ki-67 staining indices could be observed than in their primaries, even when the histological grading did not change substantially. In a cerebellar hemangioblastoma, a trigeminal neurinoma and two endotheliomatous meningiomas the fraction of stained nuclei was less than one percent; however, one recurrent transitional meningioma without any histological sign of malignancy showed a staining index of 2.4 percent. The staining indices of five brain metastases of different malignancies ranged from 1.5 percent in a malignant melanoma to 6.1 percent in bronchial carcinoma. In the majority of the cases examined, the percentage of Ki-67 labelled cells was in accordance with the histologic grade of the neoplasm. In general, there was a direct relationship between the number of stained nuclei and the frequency of mitoses (mitotic index) evaluated in hematoxylin-eosin stained frozen sections. Interestingly, the frequency of mitosis and stained nuclei were higher in tumor recurrences than in the primaries. The results of this study imply that immunohistological labelling of the proliferating cell fraction should become an important additional criterion to predict the biological behaviour of human nervous system neoplasms. PMID- 3052246 TI - Induction of antimicrobial activity by antitumor substances from pine cone extract of Pinus parviflora Sieb. et Zucc. AB - Pretreatment with two distinct antitumor substances extracted from pine cone of Pinus parviflora Sieb. et Zucc. protected mice from the lethal effects of E. coli infection. Intraperitoneal administration of these fractions transiently induced differentiation-inducing factor (DIF) with a peak at 1-2hr. The rapid decay of DIF activity from the peritoneal cavity was followed by polymorphonuclear cell (PMN) accumulation and enhancement of superoxide generation (assayed with luminol dependent chemiluminescence (LDCL] by peritoneal exudate cells. The superoxide generation by adherent cells was similarly enhanced by pretreatment, but was only 10-20% of that of the peritoneal exudate cells. Fractions that showed comparable antitumor/antimicrobial activity were also obtained from seed shells and cones of other pine trees of Japanese and foreign origin. On the other hand, a neutral polysaccharide fraction from Pinus parviflora Sieb. et Zucc. that lacked any of these activities did not induce PMN accumulation, DIF activity or LDCL generation. The results suggest a significant role in PMN activation for the expression of antimicrobial activity induced by pine cone extracts. PMID- 3052247 TI - In vitro and in vivo activity of 1-aryl-3-(2-chloroethyl) urea derivatives as new antineoplastic agents. AB - A few 1-aryl 3-(2-chloroethyl)ureas (CEU) were synthesized and screened in vitro for their cytotoxicity. Some of these derivatives were assayed for their mutagenicity, their in vivo toxicity and their antineoplastic activity. Methyl 4 (p-(3-(2-chloroethyl) ureido) phenyl) butyrate, 4-methyl and 4-tertbutyl (3-(2 chloroethyl) ureido) phenyl) butyrate, 4-methyl and 4-tert-butyl (3-(2 chloroethyl) ureido) benzene had an ID50 of 28, 20 and 4 microM respectively when tested on LoVo cells, while chlorambucil (CBL) and CCNU had an ID50 of 21 and 45 microM. These 3 chloroethyl urea derivatives were not toxic when injected i.p. at doses up to 220 mg/kg, whereas chlorambucil was already toxic at 18.5 mg/kg. The survival time of BDF1 mice bearing L1210 leukemia tumors was significantly enhanced by intraperitoneal injections of CBL and CEU. The most cytotoxic derivative (tert-butyl derivative) gave the best antineoplastic activity with a median survival time 1.77 times that of the control at 10 mg/kg/day and was not toxic, whereas CBL at this concentration enhanced survival time by a factor of 1.6 and presented important side effects. The 4-tert-butyl (3-(2-chloroethyl) ureido) benzene and the methyl 4-(p-(3-(2-chloroethyl) ureido) phenyl) butyrate showed no mutagenicity when assayed on TA-97, TA-98, TA-100 and TA-102, four strains of S. thyphimurium, while CBL had a weak effect on TA-102 and CCNU was highly mutagenic on TA-100 and TA-102. PMID- 3052248 TI - Hormonal profiles in women with breast cancer (review). AB - The literature concerning endogenous hormonal profiles in women with breast cancer and breast-cancer risk has been critically reviewed. The many published reports have been divided into 11 groups, with each group centered on a particular hypothesis that has been either explicitly formulated by the authors of the reports or perceived by other workers as a unifying hypothesis in certain studies. The hypotheses reviewed are: the adrenal androgen insufficiency hypothesis, the anovulation/luteal inadequacy hypothesis, the estriol hypothesis, the ovarian androgen excess hypothesis, the thyroid dysfunction hypothesis, the prolactin hypothesis, the estrone hypothesis, the estrogen-window hypothesis, the estrogen-excess hypothesis, the melatonin hypothesis, and the estrogen hydroxylation hypothesis. It is concluded that there remain, at present, only four viable hypotheses: the hypotheses of increased risk with adrenal androgen deficiency, ovarian dysfunction (luteal inadequacy and excessive ovarian androgen secretion), increased 16 alpha-hydroxylation of estradiol, and the hypothesis of decreased risk with pregnancy-induced lowering of prolactin levels. Adrenal androgen deficiency seems to be pertinent only in premenopausal cancer patients, and may be a genetic defect. Ovarian dysfunction seems to be pertinent to both premenopausal and post-menopausal patients and may also have a strong genetic component. Increased estradiol hydroxylation likewise seems to have a genetic component. The prolactin effect differs from the others, in that it is clearly environmental, rather than genetic, and may represent a permissive effect rather than a true risk-promoting effect. PMID- 3052249 TI - Cancer staging and survival rates: an analysis. AB - The interrelation between tumor staging and survival is an inverse one; patients with more widespread disease tend to have a shorter survival. Using this as a basis, a relationship was given between the logarithm of the percent survival at five years and the tumor stage at diagnosis. The description yielded an excellent fit for six reported neoplasms. For four other malignancies, the curves revealed a shape which resembled that of a multitarget model. The simpler description of logarithm of survival versus stage could be viewed as the limiting value of a multitarget model when the target number equals 1. PMID- 3052250 TI - Mechanistic studies on N-benzyladriamycin-14-valerate (AD 198), a highly lipophilic alkyl adriamycin analog. AB - AD 198, a novel lipophilic N-alkyl derivative of adriamycin (ADR) and a potential anticancer agent for preclinical development, was studied for its effects on the activities of DNA and RNA polymerases in vitro and its ability to bind DNA. AD 198, which contains a benzyl substituent on the glycosidic amine, was found to interact with DNA through drug-DNA binding to an extent less than its parent compound ADR as shown by fluorescent emission spectra studies. It had a preferential inhibitory effect against RNA vs. DNA synthesis in vitro by RNA or DNA polymerases from both E. coli and chicken leukemia cells. Preincubation studies indicated that AD 198 may inhibit the activity of E. coli RNA polymerase through drug-template interaction and that of leukemic RNA polymerase, which uses single stranded DNA as template, through enzyme inactivation. PMID- 3052251 TI - Vitamin C, vitamin E and cancer (review). AB - The influences of vitamin C and vitamin E on cancer reported in the literature are reviewed. Several correlational studies and case-control studies suggest that the consumption of vitamin C-containing foods is associated with lower risk for certain cancers, particularly gastric and esophageal cancer. No definite links between dietary vitamin E and human cancer have been demonstrated. Animal and in vitro studies have shown that vitamins C and E can effectively inhibit the formation of carcinogenic nitrosamines. However, animal studies examining the effects of these two vitamins on other chemically-induced cancers are not conclusive. Vitamin C supplementation has been reported to inhibit skin, nerve, lung and kidney carcinogenesis. Vitamin E has been shown to inhibit skin, liver, oral, ear duct, and forestomach carcinogenesis; and to enhance, to have no effect on, or to inhibit mammary gland or colon carcinogenesis, depending upon the method of administration, the level of dietary selenium or fat, and the species and strain of animals used. Both vitamin C and vitamin E can inhibit mutagenesis and carcinogenesis in vitro. Each of the vitamins has been shown to inhibit tumor cell growth and carcinogen-induced DNA damage. The mechanism of action of the two vitamins against carcinogens is not clearly understood. Several suggested mechanisms of action include modification of the metabolism of polycyclic hydrocarbons, reduction of mutagenic activity and reaction with genotoxic free radicals. It is concluded that the potential usefulness of vitamin C and vitamin E in the prevention and treatment of cancer should not be ignored because under certain experimental conditions these two vitamins exert inhibitory effects on chemical carcinogenesis. More carefully standardized and controlled experiments are required to adequately evaluate this potential. PMID- 3052252 TI - Prognostic factors in Hodgkin's disease: implications for modern treatment (review). AB - This review outlines the major prognostic factors as derived from the analysis of recent prospective trials. Disease extent, systemic symptoms, age, sex, and achievement of complete remission lasting longer than 12 months following chemotherapy, as well as certain treatment-related complications (e.g. acute leukemia), constitute the major variables affecting survival. Bulky lymphoma and inadequate primary irradiation are factors which have influence on relapse-free but not necessarily on total survival. Recent reports provide no evidence that minimalizing treatment (except salvage treatment), will demonstrably reduce treatment-related complications. Optimal treatment, giving patients the best chance to enter first durable complete remission, still seems to represent the best strategic approach. However, in given patient subsets, the impact of various treatment strategies on the 5-, 10-, and 15-year results is now being balanced against delayed morbidity, such as organ damage and second malignancies, produced by the intensity of treatment or the prolonged delivery of certain drugs. PMID- 3052253 TI - Inhibition of proteinase-like peptidase activities in serum and tissue from breast cancer patients. AB - The inhibitory profiles of several proteinase-like peptidases active on synthetic peptide (MCA) substrates, present in sera and 100,000g supernatants of malignant tissue from patients with breast cancer, have been studied using a series of known inhibitors including epoxysuccinyl peptides (E-64, Ep-475), Z-Phe-Phe diazomethane, PMSF, iodoacetamide, 1-10-O-phenanthroline, leupeptin, aprotinin, elastatinal and alpha 2-macroglobulin. While in general the inhibition profiles confirmed reported substrate specificities some anomalies were observed. In particular, the serum activities on two cathepsin B substrates were unaffected by specific cysteine proteinase inhibitors and in breast tissue only 20-37% of activity towards these two substrates was apparently due to the presence of endopeptidases. However, the potent inhibition of other proteinase-like activities by the epoxysuccinyl peptides and leupeptin, or similar inhibitors, may be useful agents in the study of methods of combating tumour spread. PMID- 3052254 TI - Endocrine epidemiology of male breast cancer (review). AB - Male carcinoma of the breast is not a common clinical problem, but is of interest in the wider context of female as well as male breast cancer etiology. This review discusses recognized and suspected epidemiology associations of male breast cancer, and relates these to potential endocrine factors. It is concluded that altered estrogen production and metabolism, perhaps in part related to body weight in early adulthood, may prove to be the major hormonal involvement in male breast cancer. Future studies should incorporate the more recently developed approaches to investigating estrogen metabolism and bioactivity, and include familial benign and malignant breast disease. PMID- 3052255 TI - Phenotypic and genetic alterations in pre-cancerous cells in the colon. AB - Cell dysplasia in polyps and in ulcerative colitis are thought to be the pre cancerous lesion leading to invasive colon cancer. Many polyps and dysplastic lesions in ulcerative colitis have phenotypic changes (blood group antigen, cytokeratins, CEA, TAG-72.3 antigen expression) and genetic changes (c-K-ras mutation, enhanced c-myc expression and pp60c-src activity) which are characteristic of invasive cancers. Thus, these early pre-cancerous lesions may be a late stage in the genetic evolution of colon cancer. PMID- 3052256 TI - The macrophage as a production site for hematopoietic regulator molecules: sensing and responding to normal and pathophysiological signals. AB - Several functional capacities of the macrophage enable it to act as an "administrator" cell for normal and pathophysiological hemopoietic regulation. Its capacity of sensing and responding to physiso-chemical, cellular and humoral signals indicates that it can regulate myelomonocytopoiesis and erythropoiesis. This occurs by modulating colony stimulating factor and erythropoietin production in response to lactoferrin and oxygen tension respectively. Detection of erythropoietin gene expression in macrophages, both in vitro and in vivo, implies that the macrophage is an "active" member of the hemopoietic cellular microenvironment. Since a subpopulation of macrophages is responsible for this function, a model is proposed in which other hemopoietic regulator molecules may be produced by distinct subpopulations of macrophages under steady-state conditions. PMID- 3052258 TI - Oncogene transformation and the metastatic phenotype. AB - In this review, we first discuss some of the experimental parameters that need to be considered in assessing the contribution of various oncogenes to tumor metastasis. We discuss the requirement for a number of in vivo assays to measure different aspects of metastatic ability and we describe a novel metastasis assay, which we have developed, in the naturally immune-deficient chick embryo. Other factors capable of influencing the effects of oncogenes in experimental studies of metastasis, including oncogene activation and the differentiated status of the recipient cell, are also examined. We present some of our experimental results analyzing the ability of two oncogenes, src and ras, to convert cells to a metastatic phenotype. We speculate that a major mechanism by which some oncogenes promote metastatic ability is by subverting a signal transduction process, resulting in activation of a set of genes, some of which appear to promote metastatic ability. Finally, we discuss the need for additional information on the contributions of oncogenes to tumor progression and metastasis in both experimental systems as well as in clinical tumors. PMID- 3052257 TI - Possible role of the first intron of c-H-ras in gene expression: anti-cancer elements in oncogenes. AB - When members of ras gene family, which encode structurally related proteins of approximately 21,000 daltons (p21), are activated by either structural mutations or enhanced promoter activities, immortalized mammalian cells can be transformed into malignant forms. The ras genes are classified as "housekeeping genes" which express relatively constantly at low levels in all tissues and throughout all developmental stages. It is therefore important to understand how regulatory, cis acting in particular, elements of the genes control their expression. We have previously reported that the principal promoter, 51 bp long or less, located around 1370 bp upstream from the first coding ATG, plays the most important role for expression, though there seem to be residual promoter activities further downstream as well. The most upstream mRNA start site is located at the 3'-end of the principal promoter, which lies directly adjacent to the homologous sequence between human and rat c-H-ras 5'-flanking regions. It is therefore proposed that the c-H-ras gene may have a dispersed promoter in which the principal promoter and subpromoters may be distributed. On the other hand, the first intron region seems to contain two sets of positive and negative elements which appear to have, respectively, a positive and negative influence on the efficiency of focus formation with the activated c-H-ras oncogene. From testing done in our laboratory, it was further revealed that these cis-acting elements in the intron affect c-H-ras gene expression at the post-transcriptional level. The middle part of the first intron was in fact shown unusually conserved between human and rat DNA sequences. Moreover, computer analyses revealed that the intron sequences of various oncogenes (both human and rodent sequences available from GenBank DNA sequence data-bank) such as ras, fos, c-myc, N-myc and int-1, are in general significantly more highly conserved when compared with sequences of non-oncogene introns. The significance of this oncogene-related conserved sequence is discussed and the proposal is made that it may function as an "anti-cancer element" or "safety valve" against gene truncation/translocational activation of oncogenes. PMID- 3052259 TI - Onco-suppressor genes and their involvement in cancer (review). AB - Onco-suppressor genes are a heterogeneous set of genes that inhibit the cancer related phenotype of cells. Because they are difficult to identify, only a few have been described. Evidence for their existence has mainly been indirect and comes from the following types of study. 1. Recessive cancer genes in higher and lower eukaryotes have been detected. If both alleles of these genes are deleted or inactivated, cancer develops. 2. Studies on cell hybrids have implicated genes which suppress various stages in the malignant conversion of normal cells e.g. immortalisation, morphological conversion and metastasis. 3. The isolation from virally induced transformants of flat, non-tumorigenic revertants in which expression of the transforming gene is not down-regulated suggests the presence of genes which suppress the effects of transformation. 4. Blocks to differentiation can be bypassed by inducing compounds or differentiation factors. 5. Studies on tumor inhibitory factors such as tumour necrosis factor and beta TGF show that they have different effects on different types of cell-acting to promote growth in some cases and inhibit it in others. 6. The discovery of cis acting negatice regulatory elements suggests that interaction of such elements with proteins may be important for control of gene expression particularly of infecting oncogenic viruses. 7. Suppression of the transformed phenotype of malignant cells by contact with normal cells that controls growth of neighboring cells. 8. The inhibition of proliferation of transformed cells by transfection with DNA from normal cells may be a useful method for cloning onco-suppressor genes. We discuss what is known and what is not known about this important class of gene. PMID- 3052260 TI - Molecular mechanisms involved in DNA repair, in gene rearrangement and in gene amplification may be considered as an integrated system in maintaining cellular homeostasis and cell survival. AB - In procaryotic and also in eucaryotic cells, several systems for repairing the DNA damage have been discovered. The mechanism and genetic control of DNA excision-repair and SOS have been extensively studied. These repair processes help prevent lesions from interfering with DNA functions or from converting into permanent mutations that cause cellular malfunctioning and cell death, which in higher organisms contribute to malignancy. However, other molecular systems can be identified which contribute to the maintenance of homeostasis and the survival of the cell. The system that maintains the DNA superstructure is one of these while gene amplification allows the cell to become resistant to a particular environmental change and survive in the presence of chemical compounds like drugs. Our purpose was to group in a major system the molecular systems which, until now, have always been considered as separate entities. The common basis of all these systems is the capacity to maintain genetic information or to permit the survival of the cell. The system, called the "Cell Survival Molecular System" (CSMS), is composed of several well known molecular systems, such as DNA repair, which are able to restore the DNA structure and genetic mechanisms. Moreover, the Cell Survival Molecular System may also give a selective advantage to the cell so that it can grow in a hazardous environment. Following on from this, the relationship between CSMS and cancer will be discussed. The development and the progression of the cancer may be the result of the growth of an "adapted cell". This may adapt to, or resist, a new or changed environment. In addition, genetic defects can occur in CSMS and these will probably give an impaired function to some systems in CSMS. These defects could be associated with inherited or familiar cancer. PMID- 3052261 TI - Chromatin motion in interphase nuclei, its modulation and its potential role in gene expression. AB - Nuclear Rotation (NR) refers to the rotatory motion of nuclei in cells in vitro, a motion measured as the displacement of nucleoli over time. NR occurs in cycling cells; however, its observation in neurons indicates that mechanisms related to mitosis are not a prerequisite. We have shown that NR includes motion of chromatin domains in addition to those represented by nucleoli, that movements are saltatory, with periods of stationarity and reversal of direction. The observation that NR occurs independently of concurrent motion of juxtanuclear, cytoplasmic structures, leads to the concept of a stationary outer nuclear membrane. Although traditionally perceived as rotation in a two-dimensional plane, NR represents a complex, three-dimensional motion of chromatin within the interphase nucleus, with nucleoli and DAPI-stained, fluorescent chromatin domains describing curvilinear trajectories extending throughout the nucleus. Based upon evidence that the rate of this motion changes with metabolic demands, we have postulated that NR functions in gene expression, by transposing chromatin domains to be transcribed to specific nuclear compartments. In a test of this hypothesis, Nerve Growth Factor (NGF), which alters gene expression, increased NR at a time post-NGF coincident with increased activity of RNA polymerases, while GABA, also postulated to alter transcription, increased NR with near instantaneous shifts of nucleolar positions within the nuclear space. The calcium ionophore A23187 and the chelator EGTA, agents which redistribute calcium (Ca), also increased NR, while additional Ca, in presence of EGTA, returned NR to control rates. It is difficult to link NR with the action of agents which alter transcription or ion balance. Nevertheless, in support of our hypothesis, available evidence indicates that agents which alter gene expression, alter NR and that they do so, probably through calcium dependent mechanisms. PMID- 3052262 TI - PA-1, a human cell model for multistage carcinogenesis: oncogenes and other factors. AB - We have developed a cell system which utilizes the human teratocarcinoma cell line PA-1, from which we have characterized four stages of tumor progression. Soon after establishment in culture PA-1 cells revert and are no longer tumorigenic in athymic nude mice. Later, PA-1 cells as they are passaged in culture, become tumorigenic at passage 100. The transition from nontumorigenic to tumorigenic is the result of the biological effects of an activated N-ras oncogene and can be reproduced by transfection of the cloned oncogene into preneoplastic PA-1 cells. Certain preneoplastic cells (prior to passage 100) in this series are susceptible to transformation by single oncogenes while others are not. In studying the basis of this susceptibility to single oncogene induced transformation we have found that somatic cell hybrids between preneoplastic cells which can suppress ras-induced transformation and ras-transformed cells are non-tumorigenic. Therefore, we believe that the progression from ras suppressing to ras susceptibility may be due to the inactivation of a trans-dominant suppressor gene. Our system has identified at least three steps which lead to tumorigenicity; establishment of growth past senesence, activation of a ras oncogene, and inactivation of an oncogene suppressor function. Further genetic alterations are necessary for tumor dissemination and metastasis. PMID- 3052263 TI - Oncogene expression and regulation in normal lymphocytes and lymphocytes from patients with autoimmune diseases. AB - The discovery of oncogenes has offered new insights into the physiology of lymphocytes. Proto-oncogenes encode proteins that are associated with the control of lymphocyte activation, proliferation and differentiation. Mononuclear cells from patients and animals with autoimmune disorders express increased quantities of oncogenes such as c-myc, c-myb and c-raf. In this review we discuss some of the cellular and molecular events associated with lymphocyte activation and the role that the oncogenes play in each of these events. The regulation of the expression of oncogenes in these cells is also reviewed. Finally, the role of the increased oncogene expression in the pathogenesis of autoimmune diseases is discussed. PMID- 3052264 TI - A possible role for cyclosporins in cancer chemotherapy. AB - It has been established for some time that cyclosporin A (CsA) can exert cytotoxic effects in T-cell neoplasms. More recently, antiproliferative effects in a variety of non-T-cell tumour cell lines have been demonstrated. Inhibition of polyamine synthesis is a possible mechanism and combination of CsA with other polyamine inhibitors may be particularly effective. The non-immunosuppressive analogue, B3-243, is at least as effective as CsA as an antiproliferative in human lung cancer cell lines. CsA potentiates the effect of a number of cytotoxic drugs (eg adriamycin, vincristine, VP16) both in vitro and in vivo. Differential potentiation in vitro is often seen in cells with acquired drug resistance compared with their parent lines. A number of non-immunosuppressive analogues of CsA also act as 'resistance modifiers' and hence the structure/activity requirements for immunosuppression and resistance modification appear to differ. PMID- 3052266 TI - A crossover trial of bromocriptine in the treatment of vascular dementia. AB - Seven patients with vascular dementia completed an 8-month randomized double blind crossover trial of bromocriptine in a dosage of up to 30 mg per day. Patients were assessed using a modified UCLA Parkinson Rating Scale of symptoms and signs, and neuropsychological testing including the Wechsler Adult Intelligence Scale-Revised, Wechsler Memory Scale, modified Thurstone Word Fluency Test, Wisconsin Card Sort, a test of visual vigilance, and a reaction time task. Subjects failed to perform significantly better on any measure while on bromocriptine, and on several measures their performance while on the drug was worse. PMID- 3052265 TI - Oncodevelopmental expression and structure of alkaline phosphatase genes. AB - Alkaline phosphatases (APs) are members of a multigene family, that in humans include four different genes. Their wide distribution in nature, ranging from bacteria to man, indicates that APs are involved in fundamental biochemical processes. Information on the primary structure of eukaryotic APs is accumulating very rapidly. There is a high degree of similarity between the eukaryotic APs and Escherichia coli AP. Structural comparisons with the E. coli enzyme have helped identify those residues that may participate in the active site pocket, as well as predict functional-structural features unique to eukaryotic APs. The general structure of the AP genes has now been revealed through the cloning of the germ cell AP gene in humans. The entire nucleotide sequence of the gene reveals the existence of 11 exons interrupted by 10 small introns. Elucidation of the mechanism of regulation and tissue-specific expression of AP genes will be highly relevant to understanding the re-expression of these enzymes in testicular and ovarian tumors. Two vitally important developmental processes, i.e., germ cell differentiation and early embryogenesis, provide experimentally accessible models to attempt to unravel the elusive function of APs. PMID- 3052267 TI - A. B. Baker, 1908-1988. PMID- 3052268 TI - Bacterial electron transport chains. PMID- 3052269 TI - Regulation and expression of the adaptive response to alkylating agents. PMID- 3052270 TI - Nuclease hypersensitive sites in chromatin. PMID- 3052271 TI - Posttranscriptional regulatory mechanisms in Escherichia coli. PMID- 3052272 TI - Biological aspects of inorganic polyphosphates. PMID- 3052273 TI - Carnitine. PMID- 3052274 TI - Cell-surface anchoring of proteins via glycosyl-phosphatidylinositol structures. AB - A class of membrane molecules has been identified whose primary translation product includes a COOH-terminal protein sequence that signals attachment of a glycosyl-phosphatidylinositol anchor at a COOH-terminal residue that is newly formed by cleavage of the signaling sequence. This class includes a wide diversity of protein types from eukaryotes at many stages of evolution. The structures of the glycosyl-phosphatidylinositol anchors are being resolved, but their functions aside from membrane attachment and dynamics remain to be determined. PMID- 3052275 TI - DNA repair enzymes. PMID- 3052276 TI - Molecular organization and function of the complement system. PMID- 3052277 TI - Interactions between deoxyribonucleotide and DNA synthesis. PMID- 3052278 TI - Zona pellucida glycoproteins. PMID- 3052279 TI - Growth factor receptor tyrosine kinases. PMID- 3052280 TI - Lens crystallins: the evolution and expression of proteins for a highly specialized tissue. PMID- 3052282 TI - DNA polymerase III holoenzyme of Escherichia coli. PMID- 3052281 TI - Tumor necrosis, cachexia, shock, and inflammation: a common mediator. PMID- 3052283 TI - Dopamine beta-hydroxylase of adrenal chromaffin granules: structure and function. PMID- 3052284 TI - Molecular and cellular biology of intermediate filaments. PMID- 3052285 TI - Amino acid biosynthesis inhibitors as herbicides. PMID- 3052286 TI - Peptide toxins from venomous Conus snails. PMID- 3052287 TI - The biology and enzymology of eukaryotic protein acylation. PMID- 3052288 TI - Viral proteinases. PMID- 3052289 TI - Hormonal regulation of hepatic gluconeogenesis and glycolysis. PMID- 3052290 TI - Glycobiology. PMID- 3052291 TI - Structure and function of bacterial sigma factors. PMID- 3052292 TI - Transcription by RNA polymerase III. PMID- 3052293 TI - Cofactor proteins in the assembly and expression of blood clotting enzyme complexes. PMID- 3052295 TI - The Insall procedure. A method of anterior cruciate ligament reconstruction. PMID- 3052294 TI - Chemical synthesis of peptides and proteins. PMID- 3052296 TI - Semisolid state fermentation of baker's yeast in an air-fluidized bed fermentor. AB - In an attempt to grow microorganisms other than fungi using a solid-state fermentation process, a model system of Baker's yeast (Saccharomyces cerevisiae) was cultured in an air-fluidized bed fermentor. A semisolid potato mixture (pretreated with alpha-amylase) was used for the substrate in this highly aerated system. The growth of Baker's yeast in this air-fluidized bed process was easily controllable and very reproducible. Once feasible moisture levels and air flow rates were determined, the independent variables studied were the amount of the enzyme used for digesting the potato starch, the size of the yeast inoculum, and the concentration of the added defined medium. PMID- 3052297 TI - Successive mutation of E. coli for improved thiophene degradation. Scientific note. PMID- 3052299 TI - Inactivation of the thiamine transport system in Saccharomyces cerevisiae with O bromoacetylthiamine. AB - We have synthesized and characterized O-bromoacetylthiamine (BrAcThiamine), a new reagent for inactivating the thiamine transport system in Saccharomyces cerevisiae. A Lineweaver-Burk plot of data from the transport kinetic measurements showed that BrAcThiamine was a competitive inhibitor of thiamine transport in S. cerevisiae with a Ki value of 0.60 microM. Incubating BrAcThiamine with yeast cells at 40 degrees C in 0.05 M potassium phosphate buffer, pH 5.0, caused concentration- and time-dependently a remarkable loss of thiamine transport activity. The inactivating reaction of yeast thiamine transport by BrAcThiamine proceeded most effectively at pH 5.0, coinciding with the optimal pH of the transport activity. Thiamine and thiamine analogs (pyrithiamine and O-acetylthiamine) protected yeast thiamine transport activity against the inactivation by BrAcThiamine. In addition, it was found that a membrane fraction prepared from yeast cells treated with BrAcThiamine had a thiamine-binding activity only 20% of that from control cells without inactivating the binding activity of the soluble fraction. These results suggest that BrAcThiamine inactivates the uptake activity by irreversible binding to the binding site of carrier protein(s) in the thiamine transport system. PMID- 3052298 TI - Influence of unsaturated fatty acid membrane component on sensitivity of an Escherichia coli fatty acid auxotroph to conditions of nutrient depletion. AB - The unsaturated fatty acid auxotroph Escherichia coli AK7 was provided with either oleic acid (cis 18:1) or linolenic acid (cis 18:3) to vary the degree of unsaturation of cell membrane lipids. The susceptibility of oleic acid- and linolenic acid-grown cells to starvation at 37 degrees C in 154 mM NaCl was compared following the decline in the number of CFU by plating the cells on agar medium. The decline in CFU was faster for linolenic acid-than for oleic acid grown cells, but it was not indicative of cell death, since culturable CFU was recovered after respirable substrate was added to the starved cell suspension. Cell envelope microviscosity (determined by fluorescence polarization) of oleic acid- and linolenic acid-grown cells was equal in the presence of a respirable substrate, but in its absence the microviscosity of linolenic acid-grown cells was lower than that of oleic acid-grown cells. The results suggest that cell envelope microviscosity is an important factor in determining the sensitivity of E. coli to conditions of nutrient depletion. PMID- 3052300 TI - [Recent advances in retinoids studies]. AB - Retinoids are defined as the compounds which elicit the specific biological responses through binding or activating the specific receptor(s). Retinoids modulate the cellular differentiation and proliferation in many types of cells. A series of retinoidal benzoic acids, named retinobenzoic acids, have potent activities on human promyelocytic leukemia cells, HL-60, and other assay systems. Among them, Am80, AM580 and Ch55 are more active than retinoic acid in most cases. As these compounds possess quite different structures or physicochemical properties from retinoic acid or conventional retinoids, they are expected to be applied clinically for the treatments of the diseases in oncology and dermatology. Recently, progresses on the mechanistic studies on retinoidal actions have been reported. One of them is isolation and cloning of human retinoic acid-receptors and the mechanism of retinoidal action was concluded that the retinoid-receptor complex interact directly with DNA to regulate the expression of the gene, like steroid hormone. The other is the establishment of the presence of the retinoid-specific-binding protein (RSBP) by the use of our retinobenzoic acids as the probe. PMID- 3052301 TI - [Clinical studies of natural interferon alpha (HLBI) in chronic myelogenous leukemia--a multi-institutional cooperative study in Japan]. AB - Clinical trials of natural interferon-alpha (HLBI) in the treatment of Philadelphia chromosome (Ph1) positive chronic myelogenous leukemia (CML) were carried out in a cooperative study of 22 institutions in Japan. Patients with CML in chronic and accelerated phase were given intramuscular or subcutaneous injections of HLBI at the dose of 6 X 10(6) U/body on consecutive days. Of the 47 patients in this study, forty-one were evaluable for clinical effects, and 42 could be assessed for adverse effects. Among 30 evaluable patients in the chronic phase, 9 (30.0%) achieved complete remission (CR) and 20 (66.7%) partial remission (PR). Only one patient had no response (NR), and the response rate (CR + PR) in chronic phase was 96.7% (29/30). Among 11 patients in the accelerated phase, 3 (27.3%) achieved CR and 4 (36.4%) PR. The response rate in the accelerated phase was 63.6% (7/11). In all 41 evaluable patients, the response rate was 87.8% (36/41). In 5 of 13 responding patients treated for more than 6 months, cytogenetic investigation showed the decline of Ph1 positive bone marrow cells from 100% to 92-0% (mean 46%). Adverse effects such as fever (52.4%), general fatigue (35.7%), and liver dysfunction (21.4%), were observed, but they were usually mild and reversible. No patient was taken off this study because of these toxicities. The results confirm the clinical efficacy of HLBI in patients with CML in the chronic and accelerated phase. PMID- 3052302 TI - [Clinical study on the effect of natural alpha-interferon (HLBI) in the treatment of adult T-cell leukemia]. AB - The phase II trial of natural interferon-alpha (HLBI) in treatment of adult T cell leukemia was carried out as a cooperative study. Of the 24 cases which could be evaluated, 3 cases in crisis type and 5 cases in chronic type with lymphadenopathy and/or skin infiltration achieved PR, giving a response rate of 33.3%. The anti-tumor effect of HLBI for skin lesion could be assessed in 16 cases with skin infiltration, giving a response rate of 50.0% (5 CR and 3 PR) and demonstrating a high efficacy. Of the 31 eligible patients, side effects were recognised in 27 (87.1%). Major subjective and objective symptoms were fever (38.7%), fatigue (25.8%), anorexia (12.9%) and nausea (12.9%), and leukopenia (22.6%), granulocytopenia (38.7%), thrombocytopenia (38.7), elevation of GPT (12.9%) and GOT (12.9%) were observed. PMID- 3052303 TI - [Gas shadow on CT scan following treatment in metastatic brain tumor--an autopsy case]. AB - This is an autopsy case report of a 52-year-old woman with a brain abscess presenting as an intracerebral gas shadow on CT scan. She was admitted to our facility in June 1977, with disorientation and motor weakness of the right upper extremities. CT scan revealed two separate mass lesions in the frontal and occipital lobes on the left side. She was diagnosed as having metastatic carcinoma, and was treated by tumor removal, radiation therapy, and intraneoplastic local chemotherapy. Seventeen months later, she was readmitted with decreased mental activity, hemiconvulsion of the right side and high fever. CT scan revealed a peculiar round gas shadow on the left side of the temporal lobe. Ventricular drainage produced a cloudy cerebrospinal fluid with a protein concentration of more than 400 mg/dl and a leucocyte count of 138,200. An anaerobic culture of the ventricular cerebrospinal fluid revealed Escherichia coli. At that time, the lateral ventricle was irrigated several times with antibiotics. Unfortunately, however, she died one year after readmission. An autopsy was performed shortly after death, at which time serial coronal sectioning of the brain confirmed the CT scan findings presenting as an intracerebral gas shadow, and further demonstrated extension of the brain abscess from the left temporal lobe to the temporal horn of the lateral ventricle. The lesion in the left temporal lobe included yellowish pus with partially brownish capsules. A gas-containing brain abscess confirmed by CT scan and autopsy is rarely seen. The Hounsfield unit obtained from the abscess cavity by utilizing serial CT scan sometimes represents a condition requiring urgent treatment for brain abscess. PMID- 3052304 TI - Treatment of localized vitiligo by autologous minigrafting. AB - Autologous minigrafting has been reported as an effective method for repigmenting diverse types of stable leukoderma. A group of 22 patients with localized vitiligo, 17 segmental and five focal, who are under treatment with this method, are described. Thirteen patients attained a 90% to 100% repigmentation, two others achieved a partial improvement, and five patients had a positive test area indicating the possibility of repigmentation by means of this procedure. Only two patients had a negative test with minigrafts and, consequently, they were left untreated. Autologous minigrafting is suggested as an alternative for treating localized vitiligo, particularly when other medical therapeutic attempts have failed in repigmenting this often refractory condition. PMID- 3052305 TI - Complement and antibody deposition in Brazilian pemphigus foliaceus and correlation of disease activity with circulating antibodies. AB - Brazilian pemphigus foliaceus is a blistering skin disease endemic to central and southern areas of South America. In this study of skin biopsy specimens from 14 patients we present evidence that complement and immunoglobulins were present by direct immunofluorescence in the epidermal intercellular spaces in all patients. Eight of 14 patients had granular deposits of C3 in the basement membrane zone. By indirect immunofluorescence, serum samples from all 19 patients tested demonstrated the presence of circulating IgG autoantibody. Autoantibodies deposited in the intercellular spaces in titers ranging from 1:10 to more than 1:1280, and the titers drastically decreased during treatment. This is the first study to demonstrate complement deposition in the skin in Brazilian pemphigus foliaceus. PMID- 3052306 TI - Chronic intractable atopic eczema. Its occurrence as a physical sign of impaired parent-child relationships and psychologic developmental arrest: improvement through parent insight and education. AB - Atopic eczema of infancy and childhood responds readily and predictably to treatment; only a small percentage remains intractable. Lack of therapeutic response in a proportion of these patients can be attributed to dysfunctional parent-child relationships that lead to physical and emotional developmental arrest. Improvement in parent-child relationships following parental insight into their conflicted feelings permits acceptance of educational recommendations from the physician; it also allows normal development to be resumed and eczema to improve. Eight illustrative cases are reported in which aggressive dermatologic measures were combined with an approach that helped parents recognize conflict and provided education that permitted more appropriate behavioral limit setting. Rapid and sustained improvement in skin, emotional development, and social adjustment resulted. PMID- 3052307 TI - Repopulation of pigment cells in patients with vitiligo. PMID- 3052309 TI - James Spence Medallist, 1988. Professor Otto Wolff. PMID- 3052308 TI - Arciform blistering in an elderly woman. Linear IgA dermatosis (LAD). PMID- 3052310 TI - Oncogenes in malignancy. PMID- 3052311 TI - Management of constitutional delay of growth and puberty. PMID- 3052312 TI - An integrated approach to aging and depression. PMID- 3052313 TI - Multiple personality disorder: an historical perspective. PMID- 3052314 TI - [Renal transplantation using multiple independent arteries. Which revascularization technic to use?]. PMID- 3052315 TI - [Cancer of the kidney with tumor thrombosis of the inferior vena cava and right atrium]. PMID- 3052316 TI - [Obstructive uropathies: prenatal diagnosis and management]. PMID- 3052317 TI - [Bladder lymphoma associated with papillary carcinoma]. PMID- 3052318 TI - [Adrenal abscess. Presentation of a case]. PMID- 3052319 TI - [Bladder involvement in systemic amyloidosis]. PMID- 3052320 TI - Idiotypes and anti-idiotypes: what are they trying to tell us? PMID- 3052322 TI - The first international standard for antibodies to double stranded DNA. AB - This paper announces the availability of the first international standard for anti-double-stranded DNA (anti-dsDNA). The material, coded Wo/80, was obtained after recalcification of plasma taken from a patient with systemic lupus erythematosus. Vials were filled with 500 microliters serum and freeze dried. The serum contains no other autoantibodies in measurable quantities. The vials should be stored at -20 degrees C. The standard should be used for establishing national, regional, or local standards. In eight laboratories satisfactory results with the immunofluorescence technique on Crithidia luciliae were obtained; the titres varied between 1/20 and 1/640 (mean 1/160). In seven laboratories the Farr assay, with the so called 'Amersham kit', was performed. At a dilution of 1:40 a mean binding percentage of about 50% was observed. After reconstitution with 500 microliters of distilled water, the vial contains 100 IU/500 microliters or 200 IU/ml. The standard can be obtained from the custodian of WHO: Central Laboratory of the Netherlands Red Cross Blood Transfusion Service, PO Box 9190 1006 AD Amsterdam, The Netherlands. PMID- 3052321 TI - Relation of antivimentin antibodies to anticardiolipin antibodies in systemic lupus erythematosus. AB - Tests for antivimentin antibodies (AVA) were performed on 50 systemic lupus erythematosus (SLE) and 63 control sera by indirect immunofluorescence and enzyme linked immunosorbent assay (ELISA). The prevalence was significantly raised in SLE (38% and 50% of sera positive for IgM-AVA and IgG-AVA, respectively, by immunofluorescence; 36% and 64% of sera positive for IgM-AVA and IgG-AVA, respectively, by ELISA) in comparison with the control sera. A significant correlation existed between IgM-AVA, on the one hand, and anticardiolipin antibodies (ACA) and anti-single-stranded DNA (ssDNA), on the other. A stepwise principal component analysis demonstrated that IgM-AVA and IgG-AVA accounted for 71% of the total variance in SLE (50 patients x 5 parameters = total variance). Twenty ACA positive serum samples from patients with syphilis were therefore tested for the presence of AVA, but hardly any were found to be positive. IgM-AVA from patients with SLE were inhibited by cardiolipin and absorbed with ssDNA. An association between AVA positivity and arthralgia was also shown in SLE. PMID- 3052323 TI - Life threatening acute pneumonitis during low dose methotrexate treatment for rheumatoid arthritis: a case report and review of the literature. AB - A patient is described with definite rheumatoid arthritis (RA) who developed life threatening acute pneumonitis after receiving a total dose of only 12.5 mg methotrexate (MTX). This complication has been previously described, but this is probably the lowest reported dose before development of pneumonitis in a patient with RA. The possible significance of this case is discussed in the light of recent reports suggesting an increased susceptibility of patients with RA to the pulmonary toxicity of MTX. PMID- 3052324 TI - [Malaria found in patients attending a Sahelian outpatient center]. PMID- 3052325 TI - Line immunoassay: a new serologic test for malaria. PMID- 3052326 TI - Liver transplantation, including the concept of reduced-size liver transplants in children. AB - Since the establishment of a clinical program in liver transplantation in 1984, 162 liver transplants have been performed in 131 patients (78 adults, 53 children). The patient mortality rate while waiting for a suitable organ has been 8% for adults and only 4% for children (25-46% reported in the literature). The low pediatric mortality is a result of the use of reduced-size liver transplants. A total of 14 procedures have been performed in recipients whose clinical condition was deteriorating and for whom no full-size graft could be located. Of 14 children, 13 were less than 3 years of age. Patient survival is 50%, comparable to survival of high-risk recipients of full-size livers. Using reduced size liver grafting in a transplant program can lower mortality for children awaiting a transplant by overcoming size disparity. Reduced-size liver grafting will allow more effective use of donor resources and provide a potential avenue of research for organ splitting and living related donation. PMID- 3052327 TI - Experience with simultaneous pancreas-kidney transplantation. AB - With refinements in surgical techniques and increased clinical experience, there has been a resurgence of interest in vascularized pancreas transplantation. From December 1986 to April 1988, 30 whole-organ vascularized pancreas transplants with pancreatico duodenocystostomy were performed simultaneously with renal transplantation. The recipient population consisted of 20 men and ten women, with a mean age of 34.7 years (range of 25-53 years). The mean duration of insulin dependent diabetes mellitus (IDDM) was 22.6 years (range of 10-37 years). The mean pancreas preservation time was 8.7 hours (range 3-19 years). All patients were immediately insulin-independent. Simultaneous pancreas-kidney engraftment was performed to both iliac fossae via a lower midline incision (n = 28) or through a bilateral lower abdominal incision (n = 2). The mean operating time was 5.9 hours, and packed cell transfusion requirement was 1.3 units. The mean length of hospital stay was 27.4 days. Recipients averaged 2.3 admissions (1-7), with ten patients (34.4%) requiring only one hospital admission. Postoperative immunosuppression consisted of cyclosporine, prednisone, azathioprine, and Minnesota antilymphoblast globulin (MALG). A total of 49 episodes of rejection occurred in 26 patients. Actuarial patient survival rate at two years is 96.3%. The kidney and pancreas survival rates for the same time interval is 94.0% and 84.0%, respectively. Mean serum creatinine at present is 1.75 mg/dl. In conclusion, renal transplantation in concert with pancreas transplantation has a dramatic positive impact on pancreas allograft survival. Combined engraftment does not appear to jeopardize renal allograft functional survival. In view of these results, simultaneous pancreas-kidney transplantation appears to be the treatment of choice for Type I diabetic patients. PMID- 3052328 TI - Effect of propranolol administration on hemodynamic and metabolic responses of burned pediatric patients. AB - Hypermetabolism, increased heart rate, and lipolysis are responses to high catecholamine levels associated with burn injury. This study tests the hypothesis that adrenergic beta blockade in burns could reduce myocardial work, lipolysis, and negative nitrogen balance without adversely affecting cardiac or metabolic function. Eighteen patients with burns of 70 +/- 3% total burn surface area (TBSA) (Mean +/- SEM), were studied after a 5-day infusion of 2 mg/Kg of intravenous (I.V.) propranolol infusion every 24 hours without their cardiac output or resting energy expenditure being adversely reduced. Heart rate, left ventricular work, and rate pressure product were significantly reduced by 20, 22, and 36%, respectively (P less than 0.05). Plasma glucose, free fatty acids, triglycerides, and insulin levels remained unchanged. The rate of urea production, however, was significantly increased by 54 +/- 12% in fasted patients, and to a much lesser 12 +/- 2% in fed patients. The marked decrease in myocardial work afforded by propranolol administration may be of clinical benefit in the treatment of large burns. Variations in drug dosage and feeding regimens will, however, need to be perfected to limit catabolic effects. PMID- 3052330 TI - Mepacrine accumulation during treatment of chloroquine-resistant falciparum malaria. AB - Oral mepacrine dihydrochloride, 200 mg (158 mg of the base) six-hourly for five doses followed by 100 mg (79 mg of the base) eight-hourly for six days (half dosage for those less than or equal to 50 kg) was given to 21 patients with high grade chloroquine-resistant falciparum malaria in eastern Thailand. Fifteen patients (71%) had a clinical response [fever clearance time of 81 +/- 35 hours (mean +/- S.D.)] and 13 (62%) had complete clearance of parasitaemia (clearance time 92 +/- 42 hours). Two patients were cured, but 11 patients returned with recurrent parasitaemia between 11 and 40 days after starting treatment. Five patients had an R2 response and three had an R3 response. Mepacrine retains some activity against chloroquine-resistant falciparum malaria but it cannot be recommended for use in Thailand. The doses used, which are those also recommended for giardiasis, led to progressive and potentially toxic mepacrine accumulation. Further evaluation of regimens which produce safer plasma concentration profiles is needed. PMID- 3052329 TI - Intestinal gram-negative bacterial overgrowth in vivo augments the in vitro response of Kupffer cells to endotoxin. AB - A number of disease states and therapeutic maneuvers common to surgical patients can result in changes in the intestinal flora, permitting bacterial overgrowth and translocation of bacteria to gut lymphoid tissue. It is possible that these changes in gut flora increase portal levels of several factors that are capable of altering macrophage activation state, including endotoxin, lymphokines, and eicosanoids. Since Kupffer cells are directly exposed to gut factors via the portal circulation, changes in intestinal flora may influence Kupffer cell responses. Using germfree rats, it has previously been shown that the presence of gut bacterial flora is important in inducing Kupffer cells to respond to endotoxin, and that an overgrowth of gram-negative bacteria can further augment Kupffer cell responses, supporting the above-mentioned hypothesis. The current set of experiments examines how intestinal gram-negative bacterial overgrowth in normal adult rats effects the response of Kupffer cells to septic stimuli. Kupffer cells were obtained from conventional rats with induced intestinal overgrowth with Escherichia coli C25 for 2 or 7 days. After 2 days of overgrowth, Kupffer cells were only slightly less responsive to lipopolysaccharide (LPS) than control Kupffer cells. However, after 7 days of overgrowth, when placed in coculture with normal hepatocytes, Kupffer cells were significantly more responsive to LPS (p less than 0.001), inducing a greater degree of suppression in hepatocyte protein synthesis at lower LPS concentrations. When cultured alone, Kupffer cells from these animals also produced more interleukin-1 (p less than 0.002) and prostaglandin E2 (PGE2) (p less than 0.009) in response to LPS. These results show that intestinal gram-negative bacterial overgrowth in conventional rats can have direct influences on the response of hepatic macrophages to septic stimuli, and provides further support to the hypothesis that imbalances in the intestinal flora can effect the responses of immune cells in other sites of the body. PMID- 3052332 TI - The in vitro and in vivo antimalarial activity of some Mannich bases derived from 4-[7'-bromo (and chloro)-1',5'-naphthyridin-4'-ylamino]phenol and 4-(7' trifluoromethylquinolin-4'-ylamino)phenol. AB - A series of di-Mannich bases derived from 4-[7'-bromo (and chloro)-1',5' naphthyridin-4'-ylamino]phenol and 4-(7'-trifluoromethylquinolin-4' ylamino)phenol were assayed for activity against chloroquine-sensitive and chloroquine-resistant isolates of cultured Plasmodium falciparum using the inhibition of uptake of radiolabelled hypoxanthine. A number of the 4-(7' trifluoromethylquinolinyl-amino)phenols showed statistically superior activity to chloroquine and amodiaquine against both isolates. Analysis of the antimalarial activity of some of these compounds against Plasmodium berghei in mice following oral administration again demonstrated activity equal or superior to that of the established antimalarials against a chloroquine-sensitive strain, and in some cases appreciably superior activity against a chloroquine-resistant strain. PMID- 3052331 TI - Malaria infection in pregnant women in Zaire: the effects and the potential for intervention. AB - In five maternity centres in urban and rural Zaire we evaluated the maternal prevalence of Plasmodium falciparum parasitaemia and recorded fever, the frequency of abortions and stillbirths, newborn birth weights and the feasibility of delivering antimalarial chemoprophylaxis. Women in their first and second pregnancy, compared to others (greater than or equal to third pregnancies), had a higher frequency of parasitaemia (38 v. 15%, respectively, P less than 0.001), higher parasite densities (geometric mean densities 927 per mm3 v. 277 per mm3, respectively, P = 0.01), higher rates of stillbirths and low birth weight babies (24% v. 6.4%, P less than 0.001). On average, pregnant women first attended prenatal clinics in the sixth to seventh month of gestation and made three to four visits before delivery. In these areas of Zaire, antimalarial interventions during pregnancy would have the largest impact if they were targeted to women in their first and second pregnancy. In the study areas, maternal attitudes and prenatal care-seeking behaviours do not appear to be barriers to providing an antimalarial intervention. PMID- 3052333 TI - Transcapillary escape rate and capillary permeability to albumin in patients with Plasmodium falciparum. AB - The transcapillary escape rate and capillary permeability to albumin were studied in five patients with Plasmodium falciparum malaria and six control subjects by using 125I-HSA. The mean transcapillary escape rate of albumin, calculated from the slope of the plasma disappearance curve of 125I-HSA, was found to be significantly higher in the malaria patients than in the control group. As the plasma volume increased while the plasma albumin concentration decreased in these patients, this resulted in a significantly higher plasma clearance and outflux of albumin from the intravascular to the extravascular compartments. Both the effective capillary pore area per unit path length available for restricted diffusion and the specific permeability coefficient of the capillary to albumin were found to be grossly elevated in the patients' group. These findings indicated that there was an increased leakage of plasma albumin in patients with P. falciparum malaria as a result of increased capillary surface area and an increased capillary permeability to albumin. PMID- 3052334 TI - The epidemiology of shigellosis in Israel. AB - Analysis of Shigella isolates referred for serotyping to the Central Shigella Reference Laboratory has revealed a slight predominance of S. sonnei (47%) over S. flexneri (44%) in Israel. In female patients, the predominance of S. sonnei isolates was more accentuated (54 v. 37.5% for S. flexneri). On the other hand, an increased incidence of S. flexneri was observed in military personnel (59 v. 32% for S. sonnei), and even more so in the Arab population (69 v. 19%). Incidence peaked in summer (July, August) for all subgroups. PMID- 3052336 TI - Hugh Morriston Davies and lobectomy for cancer, 1912. AB - The report of a lobectomy for bronchogenic carcinoma in 1912, by Hugh Morriston Davies of London, and without precedent, describes a surgical technique strikingly similar to that of today. Unfortunately, Davies' patient died because postoperative management of the pleural space was not yet well understood. The tumor had been identified by radiographic examination and the diagnosis confirmed by cytological examination of the sputum. The operative technique included individual ligation of hilar vessels and suture closure of the bronchus, neither of which was to be reported again for more than 20 years. More effective management of the pleural space was described, without special emphasis, by Harold Brunn of San Francisco 17 years later. PMID- 3052337 TI - Coronary sinus interventions during cardiac surgery. AB - There is renewed interest in protecting jeopardized myocardium during regional and global ischemia by coronary sinus retroperfusion. Advances in catheter design and imaging techniques have made access to the coronary sinus easier and safer. Retrograde coronary sinus perfusion, aortovenous bypass, pressure-controlled intermittent coronary sinus occlusion, and synchronized retrograde perfusion have emerged as new techniques by which blood can be redirected through the coronary sinus to nourish ischemic myocardium beyond a coronary occlusion. The purpose of this review is to summarize the current results and applications of these coronary sinus interventions, and show how they can benefit the cardiac surgeon in clinical practice. PMID- 3052335 TI - Incidence and severity of acute cardiac allograft rejection with two different low-dose cyclosporine maintenance protocols. AB - Currently cyclosporine (CyA) represents the main immunosuppressive agent used after cardiac transplantation and usually is administered in combination with prednisone and/or azathioprine for prevention of graft rejection. From March, 1984, to August, 1987, 53 patients underwent orthotopic heart transplantation for terminal-stage heart disease at the Second Department of Surgery, University of Vienna. All patients received CyA in increasing dosage (3 mg/kg to 6-10 mg/kg) postoperatively according to renal function, obtaining a trough high-pressure liquid chromatographic whole-blood target level of 200 to 400 ng/ml at the end of the first week. CyA was subsequently tapered to 100 to 150 ng/ml after 6 months. From March, 1984, through April, 1986, maintenance immunosuppression was carried out with a double-drug regimen of CyA and azathioprine. Since May, 1986, a triple drug schedule was applied with CyA, azathioprine, and prednisone. Under triple drug therapy, the incidence of mild, moderate (p less than 0.0001), and severe (p = 0.05) allograft rejection proven by endomyocardial biopsy decreased significantly with a corresponding increase of absent (p less than 0.0001) rejection. Freedom from moderate, severe, and lethal graft rejection, number of rejection episodes per patient after 1 year (double drug, 1.0, versus triple drug, 2.5), and patient survival disclosed significant improvement for recipients of the triple-drug regimen. Both groups had the same incidence of infectious complications; freedom from death by infection after 1 year was 90% versus 91% (double versus triple drug, p = 0.20).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3052338 TI - In defense of the concept of biological aging measurement--current status. AB - Biological age is the objective assessment of a person's health status. Theoretically, a 'normal' person's biological age--in terms of appearance, performance, and functional capacity--should be the same as his chronological age. Many scientists have attempted to develop systems to accurately determine individuals' biological age. Typically, the approach is to select a battery of test parameters comprised of tests which correlate closely with chronological age. This approach assumes that those traits which vary most closely with age are the best indicators of the aging process. The goal has been to compare an individual to his chronological age peers to determine his relative aging status. Two papers (Costa and McCrae, 1980 and 1985) that criticize this concept and approach have heretofore gone unanswered. Lack of published dissent has caused many gerontologists to assume that Costa and McCrae are correct in their assertions that biological age cannot be measured and is not a valid concept. Consequently, some scientists have been reluctant to pursue research in this area. The purposes of this paper are: to critically evaluate the questions raised by Costa and McCrae; to reaffirm the validity of the concept of biological age; and to urge continued research in this most important subject. PMID- 3052339 TI - Measures and markers of biological aging: 'a great clamoring ... of fleeting significance'. AB - In response to Dean and Morgan (1988), we review our position on approaches to functional or biological aging. Researchers have attempted to assess an hypothesized underlying 'rate of aging' by combining information from the functioning of several different physical or psychological systems. None of these attempts has yet demonstrated success; because many different processes contribute to what we call 'aging', the concept of a single biological age is itself probably fundamentally flawed. We advocate more sophisticated interdisciplinary and longitudinal research as the best hope for understanding and ameliorating the effects of aging processes. PMID- 3052340 TI - On the ontology of biological aging. AB - The central question in the field of experimental gerontology is: what is biological aging? It is true that a living body must obey the laws of causal determinism; like all bodies it is subject to the physical and chemical laws of the phenomenal world. But there is a further aspect; as well as the finality which unites the part, the organism and the organ, there is also a metaphysical meaning at work in life. PMID- 3052342 TI - [DNA repair and its relation to cell division in E. coli]. AB - In E. coli, lesions introduced by agents such as UV radiation, chemical agents, thymine starvation, lead to the induction of a series of bacterial fonctions called the "SOS response" (DNA reparation, mutagenesis, filamentation, prophages induction etc...). Genetics and biochemical study set up the evidences of a regulated mechanism. The central effector of this mechanism is the Rec A protein. When a cell's DNA is damaged or its DNA replication is inhibited, an inducing signal is generated. The inducing signal reversebly activates a specific protease activity of Rec A which allows it to cleave the Lex A repressor. Thus inducing operons repressed by Lex A. These operons are implicated in DNA reparations and also in cellular division. A coordination between reparation and cellular division is thus established. PMID- 3052341 TI - [Louis Pasteur and the Pasteur Institutes]. PMID- 3052343 TI - [The place of mycoplasmas in scientific research]. PMID- 3052344 TI - Preoperative pulmonary evaluation. AB - Factors related to risk of perioperative pulmonary complications include site of incision, obstructive lung disease, prolonged anesthesia time, smoking history with productive cough, and obesity. Hypercapnia is a consistent indicator of high risk. There is no difference between spinal and general anesthesia with regard to risk of pulmonary complications. In patients being evaluated for lung resection, high-risk indicators include predicted postoperative forced expiratory volume in one second of less than 1000 mL, hypercapnia, severe dyspnea on exertion, or advanced age when it is associated with advanced cardiopulmonary disease. Newer methods of assessing cardiopulmonary reserve may prove useful in identifying which patients with one or more of these risk factors are suitable operative candidates. Prevention of postoperative complications in chronic obstructive pulmonary disease patients should begin in the preoperative period with discontinuation of smoking at least eight weeks before surgery and vigorous pulmonary toilet in the 48 to 72 hours before surgery. Prophylactic lung expansion maneuvers can be effective in decreasing the incidence of postoperative atelectasis in high-risk patients undergoing high-risk operations. PMID- 3052345 TI - Calf deep venous thrombosis. A wolf in sheep's clothing? AB - To determine the natural history of calf deep venous thrombosis (C-DVT), an analytic review of the 20 relevant English-language papers published since 1942 was performed. Remarkably little methodologically sound research on this subject was found. However, available evidence suggests that C-DVT propagates to the thigh in up to 20% of cases and that propagation invariably occurs before embolization. No fatal emboli were reported in patients presenting with isolated C-DVT. Traditional anticoagulation treatment with heparin sodium and warfarin sodium of symptomatic patients with C-DVT appears to prevent extension, embolization, and early recurrence. There is no convincing evidence that C-DVT leads to chronic venous insufficiency or whether the risks of anticoagulation exceed the risks of no treatment. As an option to anticoagulation, physicians may choose to follow patients with C-DVT with serial impedance plethysmography, treating only if there is evidence of proximal extension. PMID- 3052346 TI - The intravenous pyelogram in acute pyelonephritis. AB - In a cohort of 67 otherwise healthy patients with acute pyelonephritis that was severe enough to warrant hospitalization and uroradiography, 8% had a genitourinary abnormality that influenced management. Consequently, over 90% of patients had studies that did not alter their care. In an attempt to identify clinical clues that might increase specificity without compromising sensitivity of the intravenous pyelogram in acute pyelonephritis, only the fever curve was statistically useful. Confined to patients who were febrile through 72 hours of appropriate antibiotic treatment, the yield of urography in demonstrating anomalies of immediate clinical significance rose from 8% to 36%. The likelihood of an acutely important abnormality was also increased fivefold in both diabetic patients and patients with a urinary pathogen other than ampicillin-sensitive Escherichia coli, but small numbers precluded statistical significance. Bacteremia was common (27%), but not helpful other than in confirming the microbiological diagnosis. Nonacute structural abnormalities were present in 43% of the patients, three to nine times more frequently than in reported cases without upper tract infection. PMID- 3052347 TI - Heat susceptibility of bacterial enteropathogens. Implications for the prevention of travelers' diarrhea. AB - The heat susceptibility of four bacterial enteropathogens in foods and water was studied to develop effective recommendations for travelers to regions where diarrheal diseases are important health problems. All enteropathogens tested survived well in foods stored at refrigerator temperature (4 degrees C), room temperature (25 degrees C), and 50 degrees C, which is too hot to touch. Tap water had to be heated above 65 degrees C to reliably kill all bacterial enteropathogens. At 13 of the 14 tourist-oriented hotels in four countries, water from the hot water tap did not reach temperatures of 65 degrees C. The implications of this study are that food and water that are too hot to touch may still be contaminated with bacterial enteropathogens. Travelers should be advised that food, water, or beverages are safe only if they have been brought to boiling or near-boiling temperatures prior to consumption. PMID- 3052348 TI - Obstructive sleep apnea in amyloidosis treated with nasal continuous positive airway pressure. AB - Obstructive sleep apnea can occur in patients with a variety of upper airway anatomic abnormalities including macroglossia. We present a case of macroglossia secondary to amyloidosis causing obstructive sleep apnea that was successfully treated with low pressures of nasal continuous positive airway pressure (5 cm of water). PMID- 3052349 TI - Parental loss in childhood. Its effect in adult life. PMID- 3052350 TI - Arterial wall injury and proteoglycan changes in atherosclerosis. AB - The concept of injury as a mechanism leading to atherosclerosis has been fostered by numerous studies of initiating factors and by observation of the response of cardiovascular connective tissue, ie, cellular and extracellular matrix components. Carbohydrate-protein macromolecules of the extracellular matrix are a complex group of biologically important substances that play a crucial role in mesenchymal tissue repair following injury, a process needed to maintain arterial wall integrity. Of particular interest are the proteoglycans that enter into a variety of roles, from that of inhibiting atherosclerosis and helping to maintain fibrillar structures to that of taking part in lipid deposition in the development of atherosclerosis. PMID- 3052351 TI - Animal models and the study of atherosclerosis. AB - Identifications, requirements, and numerous contributions of animal models, as well as different aspects of atherosclerosis that were successfully studied in animals, are reviewed. Suggestions are made for updating the selection of specific animal models that will most satisfactorily fulfill the needs of studies in a particular problem of this disease process. Attention has been called to the importance of the type of atherosclerotic lesions and their dependence on species susceptibility and dietary influence. The role the type of lesion plays in atherosclerosis has been emphasized, especially in studies concerned with the progression and therapy of this disease process. From the variety of animals listed in this review, as well as the number of different problems already studied or awaiting further study, it appears that numerous data can be reconciled to help elucidate many problems and provide information for our better understanding of atherosclerosis. This, in turn, will allow us to make more accurate and effective recommendations leading to prevention and/or treatment of atherosclerosis. PMID- 3052352 TI - Hemodynamics and atherosclerosis. Insights and perspectives gained from studies of human arteries. AB - Atherosclerosis affects the major elastic and muscular arteries, but some vessels are largely spared while others may be markedly diseased. The carotid bifurcation, the coronary arteries, the infrarenal abdominal aorta, and the vessels supplying the lower extremities are at highest risk. The propensity for plaque formation at bifurcations, branchings, and curvatures has led to conjectures that local mechanical factors such as wall shear stress and mural tensile stress potentiate atherogenesis. Recent studies of the human vessels at high risk, and of corresponding models, have provided quantitative evidence that plaques tend to occur where flow velocity and shear stress are reduced and flow departs from a laminar, unidirectional pattern. Such flow characteristics tend to increase the residence time of circulating particles in susceptible regions while particles are cleared rapidly from regions of relatively high wall shear stress and laminar unidirectional flow. The flow patterns associated with plaque localization are most prominent during systole. Long-term consequences are therefore likely to be greatly enhanced by elevated heart rate and may exert a selective effect on the coronary arteries. The point-by-point redistribution of wall tension at regions of geometric transition has not been quantitatively related to plaque localization. Enlargement of arteries as plaques increase in size and the associated modeling of plaque and wall configuration tend to preserve an adequate and regular lumen cross section. Hemodynamic forces appear to determine changes in vessel diameter so as to restore normal levels of wall shear stress, while wall thickness architecture, and composition are closely related to tensile stress. Hemodynamic forces may also be implicated in the symptom-producing destabilization of plaques, especially in relation to wall instabilities near stenoses. The relative roles of wall shear stress, tensile stress, and the metabolism of the artery wall in the progression and complication of atherosclerosis remain to be clarified. Development of clinical techniques for relating hemodynamic and tensile properties to plaque location, stenosis, and composition should permit pathologists to provide new insights into the bases for the topographic and individual differences in plaque progression and outcome. PMID- 3052353 TI - Dietary cholesterol and human coronary heart disease. The epidemiologic evidence. AB - For decades, dietary cholesterol has been recognized as the "materia peccans" (Anitschkow) for the induction of atherosclerosis in animals. In rabbits, chickens, and monkeys, long-term feeding of small amounts of cholesterol leads to atherosclerosis despite little or no rise in serum total cholesterol, indicating an independent contribution of dietary cholesterol to atherogenesis over-and above its influence on serum cholesterol, possibly through effects on serum cholesterol fractions (eg, increased low-density lipoprotein-cholesterol and decreased high-density lipoprotein-cholesterol). In humans, ingestion of dietary cholesterol raises serum cholesterol, largely through its effect on low-density lipoprotein-cholesterol. Over the range of intake in usual American diets, this effect is substantial, eg, with 300 mg of cholesterol intake per 1000 kcal, rather than 100, serum cholesterol is on average about 6% to 7% higher, equivalent to a 12% to 14% greater risk of coronary heart disease (CHD). In international studies based on the Food and Agriculture Organization and the World Health Organization (Geneva) data, mean per capita dietary cholesterol levels are consistently related to CHD mortality rates. In addition, since 1981, four prospective within-population studies have shown that dietary cholesterol intake of individuals is significantly related to their long-term CHD risk, independent of and in addition to serum cholesterol, blood pressure, and cigarette use. On average, a 200-mg/1000 kcal higher intake of cholesterol at baseline was associated with a 30% higher CHD rate (95% confidence interval, 1.1 to 1.5). Conversely, lower intakes of cholesterol were associated with significantly lower risks of CHD, and of all causes mortality as well. For example, with 19 years of follow-up in the Chicago Western Electric Study, a 200 mg/1000 kcal habitual lower cholesterol intake was associated with a 37% lower risk of death from any cause, equivalent to a life expectancy longer by 3.4 years. The importance of a low-dietary cholesterol intake for prevention of CHD merits increased emphasis. PMID- 3052354 TI - Cholesterol vehicle in experimental atherosclerosis. A brief review with special reference to peanut oil. AB - In general, the level of unsaturation of a fat determines its effect on plasma cholesterol level-the level being lower with more unsaturated fat. However, some exceptions do exist. One such exception is cocoa butter, which is not as cholesterolemic or atherogenic for rabbits as would be expected from its level of saturation. The reason for this anomaly is believed to be the high level of stearic acid present in cocoa butter. Peanut oil is also anaomalous, being unexpectedly atherogenic for monkeys, rabbits, and rats. Randomization of peanut oil reduces its atherogenicity, suggesting that the structure of the triglyceride comprising peanut oil may affect its atherogenic potential. Another possibility is that a lectin present in peanut oil may be responsible for its atherogenic capacity. PMID- 3052355 TI - Atherosclerosis and apoprotein E. An enigmatic relationship. AB - In this article, we consider the role of apoprotein E in lipoprotein metabolism and especially in the metabolism of potentially atherogenic lipoproteins. Particular consideration has been given to three features of apoprotein E involvement in lipid cell interactions. Evidence implicating free cholesterol as a mediator of apoprotein E biosynthesis in cholesterol-loaded macrophages is presented. Experiments pointing to apoprotein E as the ligand promoting the interaction of beta-very-low-density lipoprotein (beta-VLDL) with macrophages are summarized. Finally, we describe the influence of fat and cholesterol fed to rhesus monkeys and baboons on the generation of hepatogenous (from isolated liver perfusates) VLDL enriched in cholesterol ester and apoprotein E. These hepatic VLDLs, none of which exhibits beta-electrophoretic mobility, promote cholesterol esterification in macrophages in proportion to their apoprotein E content. The complex role of apoprotein E in the genesis and reversal of atherosclerosis is briefly discussed. PMID- 3052356 TI - Pathobiology of atherosclerosis: concepts and perspectives. A festschrift in tribute to Robert W. Wissler, PhD, MD. PMID- 3052357 TI - The lasting contributions of Robert W. Wissler, PhD, MD, to pathology and to the pathobiology of atherosclerosis. PMID- 3052358 TI - Pathobiology of atherosclerosis--current concepts and perspectives. A festschrift in tribute to Robert W. Wissler. PMID- 3052359 TI - Arterial smooth muscle. A multifunctional mesenchymal cell. AB - In 1968, Professor Robert Wissler published a landmark review on the role of smooth muscle cells in atherogenesis. He suggested that in future studies more attention should be given to the cytologic and metabolic characteristics of these multifunctional mesenchymal cells. This article summarizes some of our studies inspired by these words and indicates a few of the many questions that still need to be addressed. We have shown that smooth muscle cells of different phenotypes from that expressed in normal arterial media exist in regions of diffuse intimal thickenings adjacent to atheroma and also in vessels following injury. Cells of equivalent phenotype can exist in cell culture, and we have shown that they have all the characteristics of smooth muscle in atherogenesis, ie, they proliferate in response to mitogens, readily synthesize increased amounts of extracellular matrix, and accumulate large amounts of lipid. In contrast, smooth muscle cells of the phenotype that normally exists in the unaffected artery do not express these characteristics and their major function is maintenance of vessel tone. Thus a deeper knowledge of the factors that control smooth muscle phenotype is essential to an understanding of the mechanisms underlying the pathogenesis of atherosclerosis. PMID- 3052360 TI - Origins of human atherosclerotic plaques. The role of altered gene expression. AB - The dramatic rise and equally dramatic fall in the mortality from coronary heart disease in the 20th century is only partly explained. This article reviews the development of our ideas concerning possible pathways other than lipids that might play a role in the development of human atherosclerosis alone or in combination with one or more of the usually considered risk factors. In some instances, such as that of cigarette smoking, the proposed concept regarding genetic alterations in vascular smooth muscle suggests a mechanism for development of at least some of the lesions. Recent studies have shown that an aberration in platelet-derived growth factor gene expression is unlikely to be a factor in proliferation of smooth-muscle cells. Aberrant expression of other oncogenes or some as yet unknown virus remain as possible explanations of some of the incidence of atherosclerosis and its consequent coronary heart disease. PMID- 3052361 TI - The healing of biologic and synthetic bone implants. An experimental study. AB - The aim of the present study was to investigate the osteogenic properties of different types of cancellous bone grafts inserted in large osseous defects in dogs and to compare these with those of sintered hydroxyapatite implants. Fresh cancellous autografts were rapidly revascularized and invariably induced a complete healing of the defect. Frozen and fresh cancellous allografts were largely resorbed, the latter evoking a strong antigenic response in two of the five cases. Sintered hydroxyapatite granules were largely encapsulated in fibrous tissue, neither stimulating nor inhibiting osseous ingrowth. Degradation of the hydroxyapatite implant was not observed. PMID- 3052362 TI - Ambulation in the adolescent with spina bifida. II. Oxygen cost of mobility. AB - This study was designed to determine the energy cost (measured as oxygen use) of walking and wheelchair propulsion in children aged 10 to 15 with myelomeningocele of thoracic to sacral levels, and to determine whether energy cost of mobility could be estimated from clinical measures. Oxygen consumption (measured with open circuit spirometry) and heart rate were measured during treadmill walking by 21 children, wheelchair use by eight children, and, for five children, in both modes. Speeds ranged from 27 to 134 m/min, with slopes up to 15%. Energy consumption for walking was linearly related to speed, slope, heart rate, and body weight (r = .90, p less than .001); for wheelchair propulsion, energy consumption was a linear function of speed, slope, and body weight (r = .90, p less than .001). The same linear function applied for all disabled children; maximum walk/run speed over a 30 m distance correlated highly with both maximal oxygen consumption (r = .87) and speed using 70% of VO2max (r = .82). For both wheelchair use and walking, the relative energy consumption (percentage of VO2max) was highly correlated with heart rate alone (r = .93), and the absolute level of energy consumption was highly correlated with heart rate and maximum walk/run speed (r = .89). Simple clinical measures of maximum ambulatory velocity and heart rate allow accurate prediction (r = .89) of energy consumption in all children with myelomeningocele, regardless of neurologic and functional level. PMID- 3052363 TI - The orthograde venous autograft and allograft. Presidential address. PMID- 3052364 TI - Orthotopic liver transplantation for biliary atresia. Evolution of management. AB - Forty-five patients with biliary atresia were accepted for orthotopic liver transplantation. Nine patients died awaiting transplantation, and 36 underwent transplantation. A portoenterostomy had been performed in 28 of these 36 patients, and its presence did not significantly affect the intraoperative blood loss (5.6 vs 4.1 blood volumes), the need for retransplantation (21% vs 12%), biliary complications (21% vs 12%), postoperative infections (36% vs 25%), or survival (82% vs 63%). These results indicate that early portoenterostomy is appropriate early therapy for biliary atresia; however, prompt referral to a liver transplant center for evaluation at the first sign of cholestasis is needed to attain optimal results for transplantation. Revisions of the portoenterostomy prior to transplantation did not improve the longevity of the procedure but did substantially increase complications and death after orthotopic liver transplantation. PMID- 3052365 TI - Hepatic trauma. AB - Two hundred thirty-three patients were operated on for hepatic trauma during a two-year period. There were 101 patients with stab wounds, 90 with gunshot wounds, and 42 with blunt trauma. There were 56 isolated liver injuries. Three hundred seventy-five associated injuries occurred among the remaining 177 patients. The majority of patients required only drainage. "Liver sutures" were employed in 66 patients. Only 18 patients required debridement, resection, or packing. Twenty-eight patients (12%) died. Perioperatively, 13 patients died of hemorrhage from the hepatic wound and from the associated major vascular injuries that were present in eight of the 13 cases. The remaining deaths were not primarily a consequence of the hepatic wound. Control of hemorrhage remains the dominant consideration in the treatment of major hepatic wounds. PMID- 3052366 TI - Agenesis of the gallbladder without extrahepatic biliary atresia. AB - Agenesis of the gallbladder without extrahepatic biliary atresia is a rare disorder. At the UCLA-affiliated hospitals, 12 patients were classified in the following groups: (1) multiple fetal anomaly, (2) asymptomatic, and (3) symptomatic. All four patients in the multiple fetal anomaly group died of their other congenital defects. In the three patients in the asymptomatic group, the absent gallbladder was an incidental finding at autopsy. The five patients in the symptomatic group underwent operations for symptoms suggestive of biliary tract disease, with no gallbladder found; all were symptom free postoperatively. Operative strategy should include a complete exploration, operative cholangiography, and common bile duct exploration as necessary. Possible mechanisms responsible for symptoms include primary duct stones, biliary dyskinesia, or nonbiliary disorders. Computed tomography, biliary manometry, upper gastrointestinal tract endoscopy, and endoscopic cholangiography (with or without sphincterotomy) could be employed if symptoms continue. PMID- 3052368 TI - [Platelet activating factor in allergic reaction]. PMID- 3052367 TI - Metabolic interaction between skeletal muscle and liver during bacteremia. AB - To study the effects of bacteremia on skeletal muscle leucine (LEU) metabolism, mongrel dogs were infused with normal saline or Escherichia coli (10(9)/kg). After a bolus dose (3.6 microCi), L(1-carbon 14) LEU (0.3 microCi/min) was infused directly into the isolated, constant-flow, in vivo gracilis muscle. Arteriovenous differences for amino acids, labeled and unlabeled LEU and alpha ketoisocaproic acid (KIC), and labeled carbon dioxide were measured at ten-minute intervals for one hour. Bacteremia increased the net release of amino acids and total N2 from muscle. Moreover, plasma LEU that was deaminated and released as KIC was increased, and there was also an increase in decarboxylated plasma LEU during bacteremia. Despite the marked increase in KIC release from skeletal muscle during bacteremia, arterial concentrations were not significantly different from those of controls. An unchanged arterial plasma KIC concentration associated with a marked increase in KIC released from skeletal muscle indicates an increase in LEU metabolism, most likely in the liver. Thus, the increased skeletal muscle catabolism is not a futile cycle but rather an essential event to meet the increased metabolic needs of the body during bacteremia. PMID- 3052369 TI - Total or partial hepatic assist: limitations and possibilities. PMID- 3052370 TI - Will artificial liver therapy ever become a reality? Historical aspects and future prospects of the artificial liver. PMID- 3052371 TI - Approach to hepatic assist utilized in Japan. PMID- 3052372 TI - Chronic hepatic assist. PMID- 3052373 TI - Artificial endocrine pancreas (closed-loop-system for blood sugar control in diabetes mellitus): introduction to the subject. AB - Clearly, continuous blood glucose monitoring by portable instruments is the only and absolute prerequisite for unprejudiced evaluation of the various strategies for substitution of insulin deficiency in any form of diabetes mellitus. Continuous blood or tissue glucose monitoring remain the prerequisite for reestablishing a satisfactory feedback control mechanism between insulin secretion and blood glucose concentration in any nondiabetic patient. Loss of first-phase insulin secretion produces defects in regulation of carbohydrate metabolism, as in type II diabetic human subjects. All efforts to solve this important problem are justified. PMID- 3052374 TI - Large subcortical hemispheric infarctions. Presentation and prognosis. AB - A specific form of large subcortical hemispheric infarction on computed tomography was identified in 24 of 2198 (1%) stroke registry patients. Combined with 13 cases from earlier literature reports, a characteristic neurologic picture developed. Severe face plus arm plus leg weakness at onset (76%), corticallike features of aphasia and/or contralateral neglect (68%), and premonitory transient ischemic attacks (24%) were frequent. Twenty-two patients (59%) had large vessel arterial occlusive disease. Eight patients (22%) had primary embolic occlusion in the middle cerebral artery territory. During an average follow-up of 16 months, five patients (14%) suffered recurrent stroke or death. The clinical presentation and prognostic features of this distinct stroke subtype are described. PMID- 3052376 TI - Some reminiscences. PMID- 3052375 TI - The 'Kennard effect' before Kennard. The early history of age and brain lesions. AB - The role of age in recovery of function after brain damage has been of particular interest since the mid-1930s when Kennard described sparing of motor function following brain damage in infant monkeys. In the years since her initial papers, this phenomenon has become known as the "Kennard principle." This article describes a number of observations of the Kennard principle prior to Kennard's first publication. Included are descriptions of both early animal research and neurologic cases. PMID- 3052377 TI - Caries experience in the permanent dentition of late mediaeval Scots (1300-1600 a.d.). AB - Dental caries prevalence, distribution and site of attack were in broad agreement with previous reports for Mediaeval Scots, namely that caries was principally a disease of adult life and showed a different location distribution from that of modern caries. The findings reinforce evidence that caries prevalence in Scotland was lower than in England at that period. It is suggested that reliable estimates of caries prevalence can best be made by noting: (i) individual caries experience, (ii) lesion location by tooth type and area of attack, (iii) the number of carious teeth of each type as a percentage of teeth of that type present. PMID- 3052378 TI - Histological features and in-vitro proteoglycan synthesis in the rabbit craniomandibular joint disc. AB - The biomechanical properties of this disc depend upon the composition and organization of the extracellular matrix, in which the most important elements are collagen, proteoglycan composition, and the density and orientation of the collagen fibres. The disc is composed of two thickened bands connected by a thin intermediate zone and fibrous attachment regions. Immunohistochemical analysis with monoclonal antibodies to chondroitin-6-sulphate and keratan sulphate revealed a concentration of these cartilage-characteristic glycosaminoglycans surrounding rounded, cartilage-like cells in the bands. Cells were isolated from the cartilage-like band areas and from the fibrous-attachment regions and cultured in vitro. Labelled proteoglycans synthesized by the band cells were similar to those known to be synthesized by hyaline cartilage, but the attachment region cells synthesized fibroblast-like proteoglycans. PMID- 3052379 TI - Scleral-fixated intraocular lenses. PMID- 3052380 TI - HCFA mandates preapproval for cataract surgery. PMID- 3052381 TI - Optic nerve sheath decompression. How does it work? Has its time come? PMID- 3052382 TI - Centenary celebration of Fick's Eine Contactbrille. PMID- 3052383 TI - A contact-lens. 1888 (translation) PMID- 3052384 TI - X-linked congenital stationary night blindness. Review and report of a family with hyperopia. AB - X-linked congenital stationary night blindness is almost always associated with myopia. We have reviewed all previously reported pedigrees and have found only two with patients without myopia. A recently proposed classification of night blindness includes a complete type associated with myopia and an incomplete type in which both hyperopia and myopia were found. Complete and incomplete types did not occur within the same pedigree. We report on a family in which three of the five affected members had hyperopia and could be classified as the incomplete type and in which a fourth member with myopia was more consistent with the complete type. The lack of myopia in three members of our pedigree can be explained by two hypotheses: crossing over of the night blindness and myopic genes on the X-chromosome, or an autosomal dominant hyperopic gene that masks the myopic gene. The data from our family support the first of these two hypotheses. PMID- 3052385 TI - Immunoglobulin deposition in the cornea after application of autologous serum. AB - A 47-year-old man with a history of multiple corneal allografts for recurrent herpes simplex keratitis developed a subtotal nonhealing corneal epithelial defect. The patient was treated with hourly drops of autologous serum. A ringlike infiltrate was subsequently observed, followed by reepithelialization of the graft. The patient later suffered allograft rejection of the cornea and recurrence of the epithelial defect, and a repeated penetrating keratoplasty was performed. Examination of the excised button demonstrated a total epithelial defect, changes compatible with allograft rejection, and, in addition, eosinophilic granular deposits within the superficial corneal stroma that corresponded to the "immune ring" observed clinically. Immunoperoxidase staining was positive for IgG, IgM, IgA, and kappa and lambda light chains. These pathologic changes lend credence to the hypothesis that the precorneal tear film may be a source of immunoglobulin that becomes deposited within the stroma. PMID- 3052386 TI - Optic nerve sheath fenestration in pseudotumor cerebri. A lateral orbitotomy approach. AB - In patients with pseudotumor cerebri accompanied by loss of vision, optic nerve sheath fenestration is an effective route to prompt recovery of vision. A lateral orbitotomy approach to decompression of the optic nerve is appropriate for the ophthalmologist with adequate orbital experience. A rectangular window of dura and arachnoid, measuring approximately 3 X 5 mm, is excised from the bulbous portion of the optic nerve. It is important that the arachnoid within the window is excised because an intact arachnoid is an effective barrier to cerebrospinal fluid egress. The use of operating microscope, microsurgical instrument, and microdissecting techniques are emphasized. Twenty-eight patients (40 eyes) with progressive visual loss were treated by surgical nerve sheath fenestration. A study of the indications, results, and complications of this procedure is presented in a companion article. PMID- 3052387 TI - A combined needle holder and scissors. AB - A combined needle holder and scissors was designed in two sizes, one for use in muscle surgery and oculoplastic surgery and the other in microsurgery. They have been used with suture sizes 10-0 to 6-0 and needles from 75 to 200 micron in diameter and have been found to be useful in cataract, squint, and oculoplastic surgery. PMID- 3052388 TI - Localization of fibronectin in human middle ear cholesteatoma. AB - Fibronectin was localized in human cholesteatoma tissues by immunohistochemical methods. The avidin-biotin-peroxidase complex staining method was used with specific fibronectin antibody. Fibronectin appeared to be localized in the matrix of the cholesteatoma studied, particularly on the surface of the cell membranes and the nuclei of the basal cells and in connective tissue. Fibronectin was not seen in the granular layer or in the keratin area. Fibronectin was found on the surface of granulation tissue, mononuclear cells, fibroblasts and endothelial cells of blood vessels. These findings were confirmed by the immunofluorescent staining method. Our previous study showed that fibronectin induced a migration of keratinocytes, macrophages and fibroblasts demonstrated by the Boyden's chamber chemotaxis assay. Macrophages and fibroblasts were shown to produce collagenase, a bone resorption factor, in cholesteatomatous tissue. The present study showed the presence of fibronectin in the matrix of cholesteatoma and granulation tissue, suggesting that fibronectin might play an important role in the clinical development and invasive behavior of cholesteatoma. PMID- 3052389 TI - Treatment of secretory otitis media with kampo medicine. AB - Kampo medicine is a traditional Japanese herbal medicine that is known as Sairei to and has been used to treat otitis media. This preparation was given orally for 4 weeks to 35 children with secretory otitis media (SOM). Four of 46 ears (8.7%) completely resolved while 32 ears (69.6%) showed a partial improvement without serious adverse reaction. Our findings indicate that Kampo medicine may resolve the inflammation and immune response associated with SOM. Further studies in its mechanism of action are warranted. PMID- 3052390 TI - Actinomycosis affecting the fallopian tube and ovary: report of 3 cases, with special reference to 2 cases following IUD application. PMID- 3052391 TI - Primary choriocarcinoma in the uterine cervix: report of 4 cases. PMID- 3052392 TI - Pure rhabdomyosarcoma of the corpus uteri: a case report with a review of the literature. PMID- 3052393 TI - Management of pregnancy complicated by diabetes mellitus: experience at the University Hospital, Kuala Lumpur. PMID- 3052394 TI - Outcome of epileptic pregnancy: neonatal hemorrhage due to anticonvulsant treatment of mothers during pregnancy. PMID- 3052395 TI - Modular caseload management--an alternative to traditional waiting lists. PMID- 3052397 TI - Malignant melanoma: the Australian contribution. PMID- 3052396 TI - Sir Albert Coates 1895-1977. PMID- 3052398 TI - Mitral valve replacement in children. AB - Rheumatic fever leading to advanced valvular heart disease, in adults and children, is still frequently seen in developing countries. In the period 1981 87, 1137 patients underwent open heart surgery for either repair (489 patients), or replacement (639 patients) of defective cardiac valves. The experience with 75 children who underwent mitral valve replacement among this group is reviewed. The aetiology of mitral valve disease was rheumatic in 71, and infective endocarditis in four; 85% of the children were in NYHA functional class III, and 15% in class IV. Seven children had intra-operative findings of rheumatic activity. Pure mitral regurgitation was seen in 41, while mixed mitral valve disease was observed in 34 children. Twenty-seven children underwent mitral valve replacement with Ionescu-Shiley bovine pericardial valves, and 48 with mechanical Bi-leaflet valves. The operative mortality was 9.3%, and the actuarial survival rate, calculated by the Cutler and Ederers method, was 87% at 5 years. PMID- 3052399 TI - Blunt trauma to the carotid artery. AB - Non-penetrating trauma to the internal carotid artery presenting as an immediate or delayed neurological deficit is an uncommon clinical entity. It has a high reported morbidity and mortality. Three cases are presented with long-term clinical and radiological follow-up. The mechanisms of injury, clinical features and possible treatment modalities are discussed. PMID- 3052400 TI - Some specimens from William Clift's copy of Matthew Baillie's The Morbid Anatomy of the Human Body (1799-1802). AB - Recently in reproducing Matthew Baillie's classic atlas of morbid anatomy using an almost complete set of the original drawings of William Clift an attempt was made to give a modern diagnosis of all the conditions illustrated. Some of the many memorable specimens are here described and illustrated. The Morbid Anatomy of the Human body is published by Melbourne University Press. PMID- 3052401 TI - Two cases of pseudo-aneurysm of the gastroduodenal artery. AB - Pseudo-aneurysm of the gastroduodenal artery is a rare cause of bleeding complicated pancreatitis. The use of computerized tomography and angiography lead to early diagnosis. The key to surgical treatment is arterial inflow occlusion prior to opening the aneurysm. PMID- 3052402 TI - Management of intrahepatic bile-duct injuries: presentation of two cases. AB - Blunt injury causing laceration of the intrahepatic bile-ducts is fortunately rare. Two cases are presented: neither was diagnosed until 48 h after initial presentation. Both were managed by laparotomy, placement of drains in the liver lacerations, and intravenous nutrition until the leaks had stopped spontaneously. The literature is reviewed and two other cases discussed. Such injuries, once diagnosed, should be treated by surgical placement of drains right at the site of leak. PMID- 3052403 TI - The psychiatry of leadership and the psychiatrist as leader. PMID- 3052404 TI - Resolutions of the Aerospace Medical Association from 1929-41: Part III--1937-41. AB - The resolutions of The Aerospace Medical Association (formerly the Aero Medical Association of the United States) have been obtained and reviewed in detail for the period 1929-41. They provide a perspective on what Association members considered priority matters during that period. The resolutions cover subjects such as pilot fatigue, aviation medical examiner selection, military flight pay for flight surgeons, and other important issues, including medical standards. These resolutions had a significant impact during the early growth of civil aviation, and a number still influence today's aeromedical practice as well as certain flight operations. PMID- 3052406 TI - Environmental biotechnology. Reducing risks from environmental chemicals through biotechnology. Proceedings of a conference. July 19-23, 1987, Seattle, Washington. Dedicated to Alexander Hollaender. PMID- 3052405 TI - G-induced loss of consciousness. PMID- 3052407 TI - Biotransformations of mercury compounds. PMID- 3052408 TI - Bioengineering issues related to in situ remediation of contaminated soils and groundwater. AB - This chapter presents some of the engineering challenges in biological degradation of organic contaminants in surface soils and the subsurface environment. Extraction of contaminants from the subsurface is generally costly, slow, and difficult. This has led to an interest in using in situ techniques for biodegradation of contaminants. For some contaminants that can be readily used as primary energy sources for bacteria with or without the presence of oxygen, bacterial removal is relatively simple and, in some situations, occurs naturally. In other cases, such as hydrocarbon spills, in situ treatment is still attractive, but has the added cost and difficulty of supplying required oxygen and nutrients for the fairly efficient aerobic oxidation. There are some indications that hydrocarbons may be biodegraded in the absence of oxygen; but, in these cases, the rates of degradation appear to be slow. More research in this area is desirable. The major challenge to both engineers and scientists lies in the decomposition of hazardous chemicals that appear to be transformed by the process of co-metabolism, or else are so low in concentration that they can only serve as secondary substrates. In either case, primary substrates are required to supply the major energy requirements for bacterial growth and/or for activation of enzymes necessary for the transformation. Engineering experience in the utilization of such processes for degradation of contaminants in the environment is very limited. Little is currently known about substrate interactions and how to optimize the bacterial transformations. Chemical requirements for achieving co metabolism are high. In addition, knowledge is lacking about methods for getting the correct amounts of chemicals to the locations where needed and in the form needed. The solution of these important issues presents a significant challenge to the engineering and science communities, requiring both basic laboratory studies and field demonstrations in well-characterized environments. PMID- 3052409 TI - Performance of biodegradative microorganisms in soil: xenobiotic chemicals as unexploited metabolic niches. PMID- 3052410 TI - Biodegradation of hydrocarbons in the environment. AB - Studies on the environmental fate of petroleum have demonstrated the nearly ubiquitous distribution of microorganisms that can metabolize hydrocarbons. The rates of degradation depend upon the concentrations of such microbes and upon the environmental characteristics of an oil-contaminated ecosystem. Given the appropriate environmental conditions, microorganisms effectively decontaminate, by their biodegradative metabolism, environments that have received petroleum pollutants. Higher-molecular-weight compounds, especially those with multiple condensed ring structures and with highly branched or substituted compounds, are relatively resistant to microbial attack. Despite the fact that a genetically engineered hydrocarbon degrader was the first organism ever patented and that seed cultures are produced by various commercial firms, enhanced biodegradation as a result of seeding generally has not been shown to be effective. Also, even though some anaerobes have now been demonstrated to be capable of hydrocarbon metabolism, hydrocarbons persist indefinitely in anoxic environments. Environmental modification, on the other hand, such as that achieved by aeration or fertilization with nitrogen and phosphorus, has been shown to enhance biodegradative removal of hydrocarbons. Having considered the various factors that influence the rates of hydrocarbon biodegradation, we are left with the question of what to do when environmental oil contamination occurs in order to minimize its persistence and thus its long-term effects. Clearly, treatment methods should enhance rather than inhibit the natural rates of oil biodegradation. In some cases, it is possible to modify environmental parameters to enhance rates of hydrocarbon biodegradation, but such methods are rarely undertaken. The translation of our scientific knowledge of hydrocarbon biodegradation into practical applications remains a major challenge. Specifically designed organisms are needed to degrade toxic aromatic components of refinery waste streams before environmental treatment. Specially designed reactors with specific microbial populations are also needed if oily sludges are to be degraded by biological means, either aerobically or anaerobically, in contained, environmentally safe reactors. PMID- 3052411 TI - The ecology of an anaerobic dechlorinating consortium. PMID- 3052412 TI - Tracking microorganisms and genes in the environment. AB - Proposed intentional releases of GEMs into the environment necessitate the development of appropriate methodologies for tracing organisms and their genes in various environmental samples. Studies have been conducted to determine the sensitivities and limitations of various methods for determining the fate of GEMs and their genes in the environment. Selective viable plate count procedures can be designed to detect the introduced organisms with high sensitivity; but they are restricted by potential mutations affecting the expression of the selective characteristic in the introduced organism, the occurrence of the particular selective characteristic in the indigenous organisms, and the need to culture the organism. The accuracy of this approach is greatly improved by colony hybridization procedures that use a specific gene probe to detect the introduced genes, but this approach is still only as sensitive as the plating procedure. Direct extraction of DNA from environmental samples, coupled with dot blot hybridization with radiolabeled probe DNA or solution hybridization, gives a high degree of both sensitivity and precision. This approach does not require culturing of the organism; and even if an introduced gene moves into a new organism or if the introduced organism is viable but nonculturable, the gene probe methods will detect the persistence of the introduced genes in the environment. Efficient direct DNA extraction methods have been developed and tested following in vitro experimental additions of GEMs to sediment and water samples. PMID- 3052413 TI - Environmental pollution policies in light of biotechnological assessment: Organisation for Economic Cooperation, United Kingdom, and European Economic Council perspectives. PMID- 3052414 TI - Prospects for laboratory engineering of bacteria to degrade pollutants. PMID- 3052415 TI - Constructing microbial strains for degradation of halogenated aromatic hydrocarbons. PMID- 3052416 TI - Stable inheritance of bacterial plasmids: practical considerations in the release of organisms into the environment. PMID- 3052418 TI - [DNA synthesis of epidermal basal cells as an indicator of the age of the wound]. PMID- 3052417 TI - [Comparison of benzodiazepine screening using the fluorescence polarization immunoassay (Abbott TDX) and thin layer chromatography in the area of low concentrations]. PMID- 3052419 TI - [Assessment of traumatic lesions of the corpus callosum]. PMID- 3052420 TI - [Forensic medicine in the 19th century in Poland]. PMID- 3052421 TI - [Possibilities for using rhinoscopy and otoscopy in forensic autopsy]. PMID- 3052423 TI - A novel aspect of the inhibition by arsenicals of binding-protein-dependent galactose transport in gram-negative bacteria. AB - The inhibitory effects of arsenate and arsenite on binding-protein-dependent transport systems are reconsidered. It is shown that arsenate inhibits binding protein-dependent galactose transport in proteoliposomes energized either by dihydrolipoamide and NAD+ or by a membrane potential (under conditions where ATP metabolism is not implicated); this result is in contradiction with the current interpretation of arsenate inhibition of binding-protein-dependent transport systems (which is based on ATP depletion) and can be explained by reference to the recently discovered ATP inhibition of the binding-protein-dependent galactose transport. In whole cells, the greater inhibition by arsenate of lipoamide dependent transport than of protonmotive-force-dependent transport may be explained by a modification by arsenate of the pools of several compounds metabolized by 2-oxo-acid dehydrogenases (which have been implicated in binding protein-dependent transport). The inhibition of binding-protein-dependent galactose transport by arsenite is probably linked to the inhibition by arsenite of the galactose-stimulated lipoamide dehydrogenase activity implicated in this transport and is reminiscent of the known arsenite inhibition of lipoamide dehydrogenases. PMID- 3052424 TI - Dietary fat and the diabetic state alter insulin binding and the fatty acyl composition of the adipocyte plasma membrane. AB - Control and diabetic rats were fed on semi-purified high-fat diets providing a polyunsaturated/saturated fatty acid ratio (P/S) of 1.0 or 0.25, to examine the effect of diet on the fatty acid composition of major phospholipids of the adipocyte plasma membrane. Feeding the high-P/S diet (P/S = 1.0) compared with the low-P/S diet (P/S = 0.25) increased the content of polyunsaturated fatty acids in membrane phospholipids in both control and diabetic animals. The diabetic state decreased the content of polyunsaturated fatty acids, particularly arachidonic acid, in adipocyte membrane phospholipids. The decrease in arachidonic acid in membrane phospholipids of diabetic animals tended to be normalized to within the control values when high-P/S diets were given. For control animals, altered plasma-membrane composition was associated with change in insulin binding, suggesting that change in plasma-membrane composition may have physiological consequences for insulin-stimulated functions in the adipocyte. PMID- 3052425 TI - Purification of alpha 2,6-sialyltransferase from rat liver by dye chromatography. AB - We describe a simple three-step purification for Gal-beta 1,4-GlcNAc-alpha 2,6 sialyltransferase (EC 2.4.99.1) from rat liver which uses chromatography on Cibacron Blue F3GA and f.p.l.c. It gives a highly purified (11,000-fold) enzyme in 19% yield, which is free of other sialyltransferases, CMP-NeuAc hydrolase, sialidases and proteinases. PMID- 3052422 TI - Molecular biology of tissue kallikrein. PMID- 3052426 TI - Biosynthesis of soluble carnitine acetyltransferases from the yeast Candida tropicalis. AB - Soluble carnitine acetyltransferase from Candida tropicalis is synthesized as a 76 kDa precursor, which is monomeric and possesses no or very little carnitine acetyltransferase activity. Maturation of the enzyme begins with proteolytic processing of the 76 kDa precursor to 64 and 57 kDa subunits. The processed subunits subsequently associate into two kinds of active oligomers; the 57 kDa subunits are assembled into a tetramer and the 64 kDa subunits into an octamer. Formation of these oligomers depends apparently on growth conditions, since both oligomers were present in cells grown in continuous culture, but cells grown batchwise contained only the tetrameric form of carnitine acetyltransferase. PMID- 3052427 TI - Expression of the renin gene in extra-renal tissues of the rat. AB - Expression of the renin gene in several rat organs is demonstrated by the detection of renin mRNA using a ribonuclease-protection technique. In two of these sites, the brain and the liver, renin mRNA levels are unaffected by changes in dietary salt which markedly affect renal renin mRNA levels. The findings provide the basis for an important ubiquitous local regulatory role for the renin angiotensin system extending beyond the circulation. PMID- 3052429 TI - Aberrant regulation of the metabolism of the insulin receptor in Swarm rat chondrosarcoma chondrocytes. AB - Treatment of Swarm rat chondrosarcoma chondrocytes for 3 days in media containing either non-recombinant pig or recombinant human insulin (1 micrograms/ml) increased the rate of proteoglycan synthesis approximately 6-fold compared with cells cultured in the absence of insulin. The concentrations of human and pig insulin that stimulated the cells to double their rate of proteoglycan synthesis were approximately 1 ng/ml and approximately 2 ng/ml respectively. Because physiological concentrations of insulin do not influence proteoglycan synthesis in non-transformed chondrocytes, the findings indicated a possible abnormality in the insulin-dependent regulation of the insulin receptor in these tumour cells. Like most cells, chondrosarcoma chondrocytes down-regulated their insulin receptors when incubated with insulin for 30 min. However, the number of plasma membrane and intracellular insulin receptors did not decrease when the chondrocytes were exposed to insulin chronically for 4 days. Chondrocytes were cultured in media containing 2H-, 13C- and 15N-labelled amino acids, and the heavy-isotope density-shift method was used to investigate both the rate of degradation and the rate of synthesis of the insulin receptor. Although the rate of synthesis of the receptor was slightly faster in the insulin-treated cultures, as assessed by a slightly faster rate of appearance of the 'heavy' receptor, the rate of degradation of the receptor was slower in the insulin-treated cultures. The half-lives for the 'light' receptors were approx. 18 h and 10 h for chondrocytes cultured in insulin-containing and insulin-free media respectively. These studies in vitro indicate that the apparent up-regulation of insulin receptors that occurs in this transformed cell upon long-term exposure to insulin is primarily the result of a decreased rate of receptor degradation. PMID- 3052428 TI - Physiological roles of nicotinamide nucleotide transhydrogenase. AB - From the foregoing considerations, the energy-linked transhydrogenase reaction emerges as a powerful and flexible element in the network of redox and energy interrelationships that integrate mitochondrial and cytosolic metabolism. Its thermodynamic features make it possible for the reaction to respond readily to challenges, either on the side of NADPH utilization or on the side of energy depletion. Yet, the kinetic features are designed to prevent a wasteful input of energy when other sources of reducing equivalents to NADP are available, or to deplete the redox potential of NADPH in other than emergency conditions. By virtue of these characteristics, the energy-linked transhydrogenase can act as an effective buffer system, guarding against an excessive depletion of NADPH, preventing uncontrolled changes in key metabolites associated with NADP-dependent enzymes and calling on the supply of reducing equivalents from NAD-linked substrates only under conditions of high demand for NADPH. At the same time, it can provide an emergency protection against a depletion of energy, especially in situations of anoxia where a supply of reducing equivalents through NADP-linked substrates can be maintained. The flexibility of this design makes it possible that the functions of the energy-linked transhydrogenase vary from one tissue to another and are readily adjustable to different metabolic conditions. PMID- 3052431 TI - A bacterial factor induces changes in cysteine proteinase forms in the cellular slime mould Dictyostelium discoideum. AB - The electrophoretic pattern of cysteine proteinases in axenically grown myxamoebae of Dictyostelium discoideum can be altered by the addition of either Gram-negative (Klebsiella aerogenes, Escherichia coli) or Gram-positive (Micrococcus lysodeikticus, Bacillus subtilis) bacteria to the culture. No changes occurred, however, if either yeast or latex beads were used in place of bacteria. The changes involved the simultaneous loss of proteinases characteristic of the axenic cells (the A-forms) and the acquisition of those found in cells which have been grown on bacteria (the B-forms). Using K. aerogenes the conversion was complete within 4 h. Extracellular proteinase activity was unaffected during this period. After the D. discoideum cells had been lysed, no equivalent change in proteinase band pattern could be produced either by prolonged incubation of cell extracts or by treatment with proteinases. An identical conversion could be induced in cultures of myxamoebae by a factor, cysteine proteinase converting factor (CPCF), present in the 15,000 g supernatant of a sonicated suspension of K. aerogenes. CPCF was macromolecular, as demonstrated by both ultrafiltration and gel filtration, acid-precipitable, but was soluble in ethanol or alkali. Its activity was unaffected by treatment with trypsin. The results suggested that CPCF might be a component of the bacterial cell wall, and since its activity was affected by lysozyme treatment, peptidoglycan is implicated. The results can be interpreted in terms of a novel nutrient-dependent post-translational change which affected most of the cysteine proteinases present in D. discoideum myxamoebae. PMID- 3052432 TI - Guanine nucleotide-independent inhibition of adenylate cyclase by a stimulatory hormone. AB - Stimulation of basal adenylate cyclase activity in membranes of neuroblastoma x glioma hybrid cells by prostaglandin E1 (PGE1) is half-maximal and maximal (about 8-fold) at 0.1 and 10 microM respectively. This hormonal effect requires GTP, being maximally effective at 10 microM. However, at the same concentrations that stimulate adenylate cyclase in the presence of GTP, PGE1 inhibited basal adenylate cyclase activity when studied in the absence of GTP, by maximally 60%. A similar dual action of PGE1 was observed with the forskolin-stimulated adenylate cyclase, although the potency of PGE1 in both stimulating and inhibiting adenylate cyclase was increased and the extent of stimulation and inhibition of the enzyme by PGE1 was decreased by the presence of forskolin. The inhibition of forskolin-stimulated adenylate cyclase by PGE1 occurred without apparent lag phase and was reversed by GTP and its analogue guanosine 5'-[gamma thio]triphosphate at low concentrations. Treatment of neuroblastoma x glioma hybrid cells or membranes with agents known to eliminate the function of the inhibitory GTP-binding protein were without effect on PGE1-induced inhibition of adenylate cyclase. The data suggest that stimulatory hormone agonist, apparently by activating one receptor type, can cause both stimulation and inhibition of adenylate cyclase, and that the final result depends only on the activity state of the stimulatory GTP-binding protein, Gs. Possible mechanisms responsible for the observed adenylate cyclase inhibition by the stimulatory hormone PGE1 are discussed. PMID- 3052430 TI - Glucose-, calcium- and concentration-dependence of acetylcholine stimulation of insulin release and ionic fluxes in mouse islets. AB - Mouse islets were used to define the glucose-dependence and extracellular Ca2+ requirement of muscarinic stimulation of pancreatic beta-cells. In the presence of a stimulatory concentration of glucose (10 mM) and of Ca2+, acetylcholine (0.1 100 microM) accelerated 3H efflux from islets preloaded with myo-[3H]inositol. It also stimulated 45Ca2+ influx and efflux, 86Rb+ efflux and insulin release. In the absence of Ca2+, only 10-100 microM-acetylcholine mobilized enough intracellular Ca2+ to trigger an early but brief peak of insulin release. At a non-stimulatory concentration of glucose (3 mM), 1 microM- and 100 microM acetylcholine increased 45Ca2+ and 86Rb+ efflux in the presence and absence of extracellular Ca2+. However, only 100 microM-acetylcholine marginally increased 45Ca2+ influx and caused a small, delayed, stimulation of insulin release, which was abolished by omission of Ca2+. At a maximally effective concentration of glucose (30 mM), 1 microM- and 100 microM-acetylcholine increased 45Ca2+ influx and efflux only slightly, but markedly amplified insulin release. Again, only 100 microM-acetylcholine mobilized enough Ca2+ to trigger a peak of insulin release in the absence of Ca2+. The results thus show that only high concentrations of acetylcholine (greater than or equal to 10 microM) can induce release at low glucose or in a Ca2+-free medium. beta-Cells exhibit their highest sensitivity to acetylcholine in the presence of Ca2+ and stimulatory glucose. Under these physiological conditions, the large amplification of insulin release appears to be the result of combined effects of the neurotransmitter on Ca2+ influx, on intracellular Ca2+ stores and on the efficiency with which Ca2+ activates the releasing machinery. PMID- 3052433 TI - Structural studies of proteins by high-flux X-ray and neutron solution scattering. PMID- 3052434 TI - Purification, crystallization and properties of porphobilinogen deaminase from a recombinant strain of Escherichia coli K12. AB - Porphobilinogen deaminase has been purified and crystallized from an overproducing recombinant strain of Escherichia coli harbouring a hemC-containing plasmid which has permitted the purification of milligram quantities of the enzyme. Determination of the Mr of the enzyme by SDS/polyacrylamide-gel electrophoresis (35,000) and gel filtration (32,000) agrees with the gene-derived Mr of 33,857. The enzyme has a Km of 19 +/- 7 microM, an isoelectric point of 4.5 and an N-terminal sequence NH2-MLDNVLRIAT. The substrate, porphobilinogen, binds to the active-site dipyrromethane cofactor to form three intermediate complexes: ES, ES2 and ES3. The gene-derived primary structure of the E. coli deaminase is compared with that derived from the cDNA of the human enzyme. PMID- 3052435 TI - Role of hydrogen in the activation and regulation of hydrogen oxidation by the soluble hydrogenase from Alcaligenes eutrophus H16. AB - The activation kinetics of the H2-oxidizing activity of the soluble hydrogenase from Alcaligenes eutrophus H16 were investigated. Activation with Na2S2O4 plus 101 kPa H2 resulted in a rapid increase in activity over 1 h and constant activity after 3 h incubation. Less-stable activations were achieved if enzyme was incubated with Na2S2O4 under 1 kPa H2 or 101 kPa N2. The enzyme could also be partly activated either with NADH alone or with H2 alone. The level of activity obtained with both 101 kPa H2 and NADH present was greater than that obtained with either 101 kPa H2 or NADH alone. Activation with H2 plus NADH was virtually independent of NADH concentration but highly dependent on H2 concentration. The effects of various concentrations of H2 and constant concentration of NADH on the level of activation were the same whether H2 oxidation was assayed by H2 dependent Methylene Blue or NAD+ reduction. Diaphorase activity did not require activation and was little affected by the treatments that activated H2-oxidizing activity. The results suggest that H2 plays an important role in regulating the level of H2-oxidizing activity in this soluble hydrogenase. PMID- 3052436 TI - Reversible and irreversible effects of nitric oxide on the soluble hydrogenase from Alcaligenes eutrophus H16. AB - The effects of NO on the H2-oxidizing and diaphorase activities of the soluble hydrogenase from Alcaligenes eutrophus H16 were investigated. With fully activated enzyme, NO (8-150 nM in solution) inhibited H2 oxidation in a time- and NO-concentration-dependent process. Neither H2 nor NAD+ appeared to protect the enzyme against the inhibition. Loss of activity in the absence of an electron acceptor was about 10 times slower than under turnover conditions. The inhibition was partially reversible; approx. 50% of full activity was recoverable after removal of the NO. Recovery was slower in the absence of an electron acceptor than in the presence of H2 plus an electron acceptor. The diaphorase activity of the unactivated hydrogenase was not affected by NO concentrations of up to 200 microM in solution. Exposure of the unactivated hydrogenase to NO irreversibly inhibited the ability of the enzyme to be fully activated for H2-oxidizing activity. The enzyme also lost its ability to respond to H2 during activation in the presence of NADH. The results are interpreted in terms of a complex inhibition that displays elements of (1) a reversible slow-binding inhibition of H2-oxidizing activity, (2) an irreversible effect on H2-oxidizing activity and (30 an irreversible inhibition of a regulatory component of the enzyme. Possible sites of action for NO are discussed. PMID- 3052437 TI - Nucleotide sequence of the phosphoglycerate kinase gene from the extreme thermophile Thermus thermophilus. Comparison of the deduced amino acid sequence with that of the mesophilic yeast phosphoglycerate kinase. AB - Using oligonucleotide probes derived from amino acid sequencing information, the structural gene for phosphoglycerate kinase from the extreme thermophile, Thermus thermophilus, was cloned in Escherichia coli and its complete nucleotide sequence determined. The gene consists of an open reading frame corresponding to a protein of 390 amino acid residues (calculated Mr 41,791) with an extreme bias for G or C (93.1%) in the codon third base position. Comparison of the deduced amino acid sequence with that of the corresponding mesophilic yeast enzyme indicated a number of significant differences. These are discussed in terms of the unusual codon bias and their possible role in enhanced protein thermal stability. PMID- 3052438 TI - Tissue-specific effects of starvation and refeeding on the disposal of oral [1 14C]triolein in the rat during lactation and on removal of litter. AB - 1. The effects of starvation and refeeding on the disposal of oral [14C]triolein between 14CO2 production and 14C-lipid accumulation in tissues of virgin rats, lactating rats and lactating rats with pups removed were studied. 2. Starvation (24 h) increased 14CO2 production in lactating rats and lactating rats with pups removed to values found in virgin rats. This increase was accompanied by decreases in 14C-lipid accumulation in mammary gland and pups of lactating rats and in white and brown adipose tissue of lactating rats with pups removed. 3. Short-term (2 h) refeeding ad libitum decreased 14CO2 production in lactating rats and lactating rats with pups removed, and restored the 14C-lipid accumulation in mammary glands plus pups and in white and brown adipose tissue respectively 4. Insulin deficiency induced with mannoheptulose inhibited the restoration of 14C-lipid accumulation in white adipose tissue on refeeding of lactating rats with pups removed, but did not prevent the restoration of 14C lipid accumulation in mammary gland. 5. Changes in the activity of lipoprotein lipase in mammary gland and white adipose tissue paralleled the changes in 14C lipid accumulation in these tissues. 6. It is concluded that 14C-lipid accumulation in mammary gland may not be affected by changes in plasma insulin concentration and that it is less sensitive to starvation than is lipogenesis or lactose synthesis. This has the advantage that the milk lipid content can still be maintained from hepatic very-low-density lipoprotein for a period after withdrawal of food. The major determinant of the disposal of oral 14C-triolein appears to be the total tissue activity of lipoprotein lipase. When this is high in mammary gland (fed lactating rats) or white adipose tissue (fed lactating rats with pups removed), less triacylglycerol is available for the muscle mass and consequently less is oxidized. PMID- 3052439 TI - Amino acid infusion increases the sensitivity of muscle protein synthesis in vivo to insulin. Effect of branched-chain amino acids. AB - Rates of muscle protein synthesis were measured in vivo in tissues of post absorptive young rats that were given intravenous infusions of various combinations of insulin and amino acids. In the absence of amino acid infusion, there was a steady rise in muscle protein synthesis with plasma insulin concentration up to 158 mu units/ml, but when a complete amino acids mixtures was included maximal rates were obtained at 20 mu units/ml. The effect of the complete mixture could be reproduced by a mixture of essential amino acids or of branched-chain amino acids, but not by a non-essential mixture, alanine, methionine or glutamine. It is concluded that amino acids, particularly the branched-chain ones, increase the sensitivity of muscle protein synthesis to insulin. PMID- 3052440 TI - Internally repeated sequences in ras gene products. PMID- 3052441 TI - Inhibition of xanthine dehydrogenase from Triatoma infestans by some purine analogues. AB - Xanthine dehydrogenase (EC 1.2.1.37) activity was determined in the fat body of the Chagas' disease vector Triatoma infestans using a system in which phenazine methosulphate was associated with p-iodonitrotetrazolium violet as electron acceptors for the oxidation of the substrate xanthine. Under the standard conditions used, xanthine was the substrate of choice. The apparent Michaelis Menten constant (Km) for xanthine was found to be 0.217 +/- 0.020 mM (mean +/- standard deviation of four independent determinations) in the absence of the inhibitors. In the presence of the purine derivatives used the average apparent Km varied from 0.216 mM to 0.219 mM, using the same enzyme preparation. In the presence of the inhibitors tested, competitive inhibition was observed with 8 azaxanthine (Ki = 0.160 mM), adenine hemisulphate (Ki = 0.184 mM) and 6 mercaptopurine (Ki = 0.008 mM). Noncompetitive inhibition was obtained with 8 azaguanine (Ki = 0.077 mM) most probably due to the formation of a dead-end complex between the enzyme, the substrate and the inhibitor. PMID- 3052443 TI - Insulin-induced tyrosine-phosphorylation in intact rat adipocytes. AB - Insulin-induced tyrosine-phosphorylation in intact isolated rat adipocytes was studied using immunoblotting method with antiphosphotyrosine antibodies. Insulin stimulated adipocytes were solubilized with Triton X-100. The lysate was incubated with wheat germ agglutinin, then with hydroxylapatite. Insulin stimulated tyrosine-phosphorylation of a 95 KDa protein which adsorbs to wheat germ agglutinin and appears to be the beta-subunit of the insulin receptor. Among the proteins adsorbed to hydroxylapatite, tyrosine-phosphorylation of 170 KDa and 60 KDa proteins was stimulated. 170 KDa was also stimulated by polyclonal anti insulin receptor antibodies B-10 Ig G, IGF-I and H2O2. The detection of these proteins in rat adipocytes may lead to the elucidation of a common signal transduction pathway in insulin-responsive cells. PMID- 3052442 TI - Is the level of steroid hormones in the low density lipoprotein (LDL) particles responsible for the prostaglandin I2 inhibitory potency of LDL? AB - In LDL fractions, isolated by ultracentrifugation, sexual hormones are detectable. LDL from male volunteers contained a significantly higher level of testosterone than LDL from female volunteers. This difference, however, is not responsible for the enhanced ability of LDL from men versus LDL from women to inhibit PGI2 formation. The importance of LDL testosterone is contradicted by the following results: Firstly, the PGI2 inhibitory potency of LDL is independent of the age of volunteers and the recentrifugation of isolated LDL fractions, but the level of testosterone is diminished with increasing age of volunteers and after recentrifugation. Secondly, LDL from hyperlipidemic men type II a did not inhibit PGI2 formation, but the level of testosterone in the LDL of these volunteers is significantly higher than in LDL from healthy men. PMID- 3052444 TI - Endothelin inhibits renin release from isolated rat glomeruli. AB - The effect of endothelin on renin release from isolated rat glomeruli was examined. Endothelin inhibited basal renin release in a dose-dependent manner with an IC50 of 1.0 x 10(-9) M. Endothelin also inhibited renin release stimulated by isoproterenol (10(-5) M). Nifedipine (10(-5) M), a calcium channel blocker, induced an increase in renin release. Endothelin did not affect this nifedipine-induced renin release. These results suggest that endothelin inhibits renin release via a calcium entry mechanism and increases intracellular calcium. PMID- 3052446 TI - Specific guanine nucleotide binding by membranes from Cucurbita pepo seedlings. AB - A microsomal membrane preparation from hypocotyls of dark-grown Cucurbita pepo L. (zucchini) seedlings contains specific high-affinity binding sites for the non hydrolyzable GTP analog guanosine 5'-[gamma-thio]triphosphate (GTP-gamma-S). Both the binding affinity and the pattern of binding specificity for GTP and guanine nucleoside triphosphate analogs are shared with the more thoroughly characterized animal G-proteins that are known to be involved in signal transduction. The sensitivity of GTP-gamma-S binding to Mg+2 ions and temperature was similar to that reported for rabbit liver G-protein, although the plant complex dissociated more readily. GTP-gamma-S could be recovered unchanged from the binding complex. Proteins (Mr 33 and 50 kDa) present in zucchini membrane preparations were revealed by immunoblotting with antiserum specific for the alpha subunit of platelet GS. These may be homologous to animal G-proteins. PMID- 3052445 TI - Release of 7-alkylguanines from haloethylnitrosourea-treated DNA by E. coli 3 methyladenine-DNA glycosylase II. AB - Previous studies have related DNA modification by the haloethylnitrosoureas to their antitumor activity. Repair of this damage, particularly by O6-alkylguanine DNA alkyltransferase, has been linked to tumor resistance by several previous investigations. We report here that E. coli 3-methyladenine-DNA glycosylase II can also remove several of the DNA modifications caused by the haloethylnitrosoureas. 7-Chloroethylguanine, 7-hydroxyethylguanine, and diguan-7 ylethane are all released into the supernatant from DNA modified by N-[2 chloroethyl-1,2-14C]-N'-cyclohexyl-N-nitrosourea. Release of diguan-7-ylethane is of particular interest since this entity evidently represents a DNA intrastrand cross-link. If a similar activity is present in mammalian cells, it might be an important source of resistance to the therapeutic action of the haloethylnitrosoureas. PMID- 3052447 TI - The transcription of the interleukin 1 beta gene is induced with PMA and inhibited with dexamethasone in U937 cells. AB - Interleukin 1 beta mRNA was induced with phorbol ester, not LPS, in the human histiocytic lymphoma cell line U937, but interleukin 1 alpha mRNA was not induced. Nuclear run-on analysis showed that phorbol ester induced the transcription of interleukin 1 beta but did not induce it in the presence of cycloheximide. This indicates that the induction of the transcription with PMA requires de novo protein synthesis. Dexamethasone, an immunosuppressive and anti inflammatory agent, decreased the level of interleukin 1 beta mRNA induced with phorbol ester. The transcriptional analysis showed that dexamethasone inhibits the transcription of the interleukin 1 beta gene. PMID- 3052448 TI - HIV-1 protease specificity of peptide cleavage is sufficient for processing of gag and pol polyproteins. AB - The mature proteins of retroviruses originate as a result of proteolytic cleavages of polyprotein precursors. Retroviruses encode proteases responsible for several of these processing events, making them potential antiviral drug targets. A 99-amino acid HIV-1 protease, produced by chemical synthesis or by expression in bacteria, is shown here to hydrolyze peptides corresponding to all of the known cleavage sites in the HIV-1 gag and pol polyproteins. It does not hydrolyze peptides corresponding to an env cleavage site or a distantly related retroviral gag cleavage site. PMID- 3052449 TI - Proteolytic activity in the insulin receptor. AB - When a highly purified preparation of rat liver insulin receptor is incubated with trypsin, the receptor develops hydrolytic activity towards N alpha-benzoyl-L arginine ethyl ester, N alpha-p-tosyl-L-arginine methyl ester, and N alpha benzoyl-DL-arginine-p-nitroanilide, (compounds which are synthetic substrates of trypsin). The activity toward N alpha-benzoyl-DL-arginine-p-nitroanilide is inhibited by soybean trypsin inhibitor but not by N alpha-p-tosyl-L-lysil chloromethyl ketone. These data are consistent with the presence of proteolytic activity in the insulin receptor specific for the bonds whose carbonyl functions are provided by arginine but not by lysine. Furthermore we found that the esterase activity toward N alpha-benzoyl-L-arginine ethyl ester in the presence of trypsin is enhanced by insulin, and that the concentration of insulin that produced the half maximum stimulation is of the same magnitude as the dissociation constant for the soluble receptor. These data suggest that the insulin receptor is a zymogen, activated by trypsin, and based on its specific activity, may be the protease which releases a peptide chemical mediator of intracellular action of insulin. PMID- 3052450 TI - Does endothelin mobilize calcium from intracellular store sites in rat aortic vascular smooth muscle cells in primary culture? AB - In the presence of endothelin, there was a rapid increase in the 45Ca++ efflux from primary cultured rat vascular smooth muscle cells, both in physiological salt solution and in calcium free medium containing 2 mM EGTA. The 45Ca++ influx was not affected. The endothelin-induced, transient increase in cytosolic calcium concentration is probably mainly due to release of calcium from the intracellular store in vascular smooth muscle cells. PMID- 3052451 TI - A novel action of activin A: stimulation of insulin secretion in rat pancreatic islets. AB - The present study was conducted to examine an action of activin A on insulin secretion from rat pancreatic islets. In a batch incubation system, activin A stimulated insulin secretion in a dose-dependent manner at concentrations higher than 1 nM. Furthermore, activin A greatly potentiated glucose-induced insulin release. When islets were perifused with 1 nM activin A, insulin secretion was barely affected in this system. However, the insulin response to 16.7 mM glucose was greatly enhanced. Both the first and second phases of insulin response were enhanced by 1 nM activin A. These results indicate that, in addition to its known actions on pituitary-gonadal and hematopoietic systems, activin A modulates the function of pancreatic islets and stimulates insulin secretion. PMID- 3052452 TI - Slipped DNA structures within the enhancer region of the Moloney murine leukemia virus. AB - We have examined the S1 nuclease sensitivity of supercoiled plasmids harboring the Moloney Murine Leukemia Virus (MoMuLV) long terminal repeat (LTR). S1 sensitivity was found within the LTR enhancer direct repeats. Transformation of E. coli DH5 cells with a construct containing most of the MoMuLV LTR yielded the precise deletion of one direct repeat and loss of S1 sensitivity. The dependence of S1 sensitivity on the presence of both direct repeats, together with the exact excision of one direct repeat by E. coli, suggests the presence of slipped DNA within the enhancer. Such structures may represent targets for effector proteins which mediate vital functions during viral propagation. PMID- 3052453 TI - 15-Hydroxyeicosatetraenoic acid dehydrogenase activity in microsomal fraction of mouse liver homogenate. AB - The microsomal fraction of mouse liver homogenate showed NAD(P)-dependent dehydrogenase activity involved in the conversion of 15-hydroxyeicosatetraenoic acid to 15-ketoeicosatetraenoic acid, which was determined quantitatively by HPLC assay. This enzyme, tightly bound to membranes and relatively stable, possessed apparent values of Km of 8.3 microM and Vmax of 2.8 nmoles/mg.min in the oxidation of 15-HETE, and gave an optimum pH of 9.8. Additionally, the enzyme, not susceptible to the inhibition by indomethacin and showing a similar cosubstrate specificity between NAD and NADP, utilized other hydroxylated eicosanoids as substrates, based on HPLC analyses. PMID- 3052454 TI - Insulin and glucose modulate protein kinase C activity in rat adipocytes. AB - In the presence of 1 mM glucose, insulin (10 ng/ml) increases both catalytic and receptor-binding properties of adipocyte cytosolic protein kinase C (PKC). Preincubation of adipocytes with 10 mM glucose raises basal PKC catalytic activity and prevents further stimulation of this enzyme by insulin. The effect of hyperglycemia is likely to be mediated by direct conversion of glucose into diacylglycerol. Thus, an incorporation of 14C-glucose into diacylglycerol is enhanced 10-fold in the presence of 10 mM glucose. These observations indicate that, in normal adipocytes, both insulin and glucose activate PKC; hyperglycemia eliminates the ability of insulin to stimulate this enzyme, thereby interfering with insulin action. PMID- 3052455 TI - Characterization of hog kidney renin-binding protein: interconversion between monomeric and dimeric forms. AB - A radioimmunoassay for hog kidney renin-binding protein (RnBP) was developed. Using this assay method, we investigated the properties of hog kidney RnBP. The lower limit of detection was 24 fmol RnBP. The molecular weight of RnBP in hog kidney extract, as well as the purified RnBP, was estimated to be 65,000 by gel filtration on Ultrogel AcA 44. When the purified RnBP was treated with N ethylmaleimide (NEM) or 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB), the molecular weight was reduced to 38,000. DTNB-treated RnBP was reconverted to the 65,000-dalton species with dithiothreitol. Cross-linked high molecular weight species of RnBP were produced by the reaction of native RnBP with dimethyl suberimidate, but formation of such species was much less with NEM-treated RnBP. These results suggest that the native RnBP exists as a dimeric form and dissociates to a monomeric form by sulfhydryl-alkylating or -oxidizing reagent. It was shown from analysis of amino acid composition of S-carboxymethylated RnBP and titration of sulfhydryl groups of native and NEM-treated RnBP with DTNB that native RnBP contained twelve cysteine residues and that three cysteine residues were alkylated by NEM under the conditions employed. PMID- 3052457 TI - Oral application of insulin encapsulated liposomes. AB - 125Iodine labelled insulin encapsulated liposomes were introduced orally to rats. After different intervals of feeding, blood from the portal vein and heart was collected. The plasma was passed through a Sepharose 2B column to establish the presence of liposomes or intact hormone. Liposomal and hormonal fractions from the column loaded with plasma from the portal vain contained radioactivity. It was found that approx. 60% of the loaded radioactivity was associated with the liposomes whereas 20% of the loaded count was found with the hormonal fractions. It is noted that undegraded protein is not detected in portal plasma from rats fed with free insulin. When plasma from the heart of rats fed with liposomal insulin was passed through Sepharose 2B column, neither liposomes nor insulin was detected. PMID- 3052456 TI - Effect of various oxygen free radical scavengers in preventing tissue injury caused by Escherichia coli in pyelonephritic mice. AB - Reactive oxygen species have been found to be responsible for the tissue injury caused in experimental pyelonephritis in mice. The extent of lipid peroxidation (as assayed by malondialdehyde formation) was found to be increased significantly (p less than .001) in the infected group as compared to the normal mice. Superoxide dismutase and catalase (oxygen free radical scavengers) showed a significant decrease (p less than .001) in the extent of lipid peroxidation even in the presence of infection. Dimethyl sulfoxide, a hydroxyl ion scavenger, was however found to be effective only at 4 and 7 days postinfection (p less than .001). Allopurinol, an inhibitor of xanthine oxidase, did not significantly (p greater than .05) inhibit the formation of lipid peroxides, even upto 7 days postinfection. There was a significant decrease (p less than .05) in the activities of renal brush border membrane enzymes used as markers of renal tissue damage (i.e. alkaline phosphatase, leucine amino-peptidase and gamma-glutamyl transpeptidase) in the infected group as compared to the normal group. In the presence of superoxide dismutase, dimethylsulfoxide and catalase except allopurinol, the activities of all the enzymes but maltase were found to be increased significantly (p less than .05) as compared to the infected group. There was a significant increase (p less than .01) in the bacterial count in the presence of superoxide dismutase and DMSO in infected mice as compared to the infected control mice. However, no significant difference was observed in the catalase and allopurinol treated groups. PMID- 3052458 TI - Effect of prolonged administration of cyclosporin A on (pro)insulin biosynthesis and insulin release by rat islets of Langerhans. AB - One group of rats received, an oral dose of 25 mg/kg body weight of Cyclosporin A daily for 21 consecutive days in olive oil, whereas the control group received an equal amount of the vehicle for the same period. On the 22nd day animals selected at random from both the groups were subjected to glucose tolerance (1.5 g/kg body weight/oral). The blood sugar values indicated glucose intolerance in experimental rats compared to vehicle-treated rats. The remaining animals were killed, and the pancreases were separated for islet isolation. After incubation of islets for 3 hr at 37 degrees with different glucose challenges, a highly significant (P less than 0.001) lowering of (pro)insulin biosynthesis, significant decrease in IRI release as well as significant inhibition in activities of acid phosphatase and cathepsin B was seen. PMID- 3052459 TI - Taurine binding to the purified insulin receptor. AB - Taurine (2-aminoethanesulfonic acid) was shown to bind specifically and reversibly to the purified human insulin receptor. While insulin binding to the purified insulin receptor exhibited characteristic negative cooperativity and an apparent dissociation constant (Kd) of 1.2 X 10(-9) M, taurine binding was shown to exhibit positive cooperativity and had a lower affinity for the insulin receptor. The apparent Kd for taurine binding to the purified insulin receptor was calculated to be 130 X 10(-9) M and the maximum number of binding sites (Bmax) was 1.6 nmol/mg receptor protein. Chromatographic data demonstrated that taurine binds to the 138,000 molecular weight subunit of the insulin receptor. Taurine binding to the receptor protein was displaced by either taurine or insulin. Anti-human insulin receptor sera prevented insulin or taurine from binding to the receptor. Taurine binding to the protein was pH dependent, and sulfur-containing taurine analogues were able to displace taurine from the insulin receptor. These data supported our previous in vivo and in vitro observations that the hypoglycemic properties of taurine appear to be mediated through an interaction of taurine with the insulin receptor. PMID- 3052460 TI - A novel autoantibody causing a peripheral fluorescent antinuclear antibody pattern is specific for nuclear pore complexes. AB - We characterized serum from a patient with polymyositis, and found that it produced a peripheral (rim) fluorescent antinuclear antibody pattern on rat liver substrate. Indirect immunofluorescence analysis revealed a punctate pattern at the nuclear surface of PtK2, BHK-21, and HEp-2 cells. This pattern was still present after sequential extraction in situ with non-ionic detergent, DNase, RNase, and high ionic strength buffer (2M NaCl). Immunogold electron microscopic localization was specific for nuclear pore complexes. By immunoblot analysis, the antigens were polypeptides of 200 kd and 130 kd that were enriched in the nuclear fraction. PMID- 3052461 TI - Comparison of the endotoxin-binding capacity of human transferrin and a human applicable immunoglobulin preparation. AB - Data obtained by in vitro experiments concerning the binding kinetics of the transferrin-endotoxin interactions are provided. The separation of protein-bound and free tritiated endotoxin of E. coli O 111:B4 is achieved by a simple precipitation method using ethanol. The main features of this reaction are the velocity of the saturation--saturation is reached after 2-3 min--and the requirement of divalent cations. EDTA (ethylenediaminetetraacetic acid) strongly inhibits the binding reaction of endotoxin to transferrin. The specificity of this interaction as well as the physiological importance can be demonstrated by the replacement of the radioactively labelled lipopolysaccharide preparation of E. coli 0 111:B4, by non-radioactive preparations of Salmonella minnesota, Serratia marcescens, E. coli 0 55:B5 and E. coli 0 111:B4. The described ethanol precipitation procedure for separating protein-bound and free endotoxin was applied to a commercially available immunoglobulin preparation. Although this preparation is known to exhibit high antibody titers against the lipid A moiety of lipopolysaccharides, no specific binding with the intact lipopolysaccharide can be found. PMID- 3052462 TI - [Modification of mucociliary clearance by a combination of theophylline with ambroxol and of ambroxol in monotherapy]. AB - Influence on Mucociliary Clearance by a Theophylline-Ambroxol Combination and by Ambroxol in Monotherapy. In a controlled randomised double-blind study the influence of theophylline + ambroxol (TA) and ambroxol (A) on lung function and mucociliary clearance was investigated. 19 patients with chronic obstructive lung disease were treated after a 5-day wash-out period for 5 days with TA (700 mg theophylline, 60 mg ambroxol/d) or A (60 mg ambroxol/d), respectively. Measurements were done on the 5th and 10th day. Under the treatment with TA a marked improvement of lung function was observed. The mucociliary clearance improved under treatment with TA and A, with TA being significantly better compared with treatment group A. PMID- 3052464 TI - Validation of biotechnological production processes. AB - Due to the biological synthesis of biotechnologically produced pharmaceuticals the product quality and safety of the drug is influenced by various factors. The correct nucleotide sequence and stability of the host cell/vector system provide the corresponding amino acid sequence of the protein. The posttranslational processing of the protein requires a well characterized production cell line. Suitable equipment for fermentation allowing a sterile production of the producing monoculture and consistent conditions are the basic requirements for the validation of the fermentation process. A constant specific productivity is one of the major criteria for the reproducibility of the production. For the validation of recovery and purification it is necessary to examine yield after each process step, product quality before and after each single process step and purification factors for removal of contaminating proteins, nucleic acids and potential viruses. In addition to the validation of the entire production process, reproducibility of quality of the formulated product has to be determined by a number of protein analytical, immunological and biochemical test methods concerning the identity, purity, safety and potency of the drug. PMID- 3052463 TI - Recombinant hemopoietic growth factors. Molecular cloning and potential therapeutic applications. AB - In analogy with data obtained from other mammalian species it appears that the majority of physiologically relevant human hemopoietic growth factors (e.g. erythropoietin and colony stimulating factors) has been characterized in biological systems, defined by biochemical methods and manufactured by molecular cloning of the corresponding genetic elements and expression in suitable biological systems. A review of the present situation of production of recombinant hemopoietic growth factors is presented and potential therapeutic applications of these factors will be discussed. In addition some preliminary clinical results, due to the medical application of recombinant hemopoietic growth factors, will be described. PMID- 3052465 TI - Inside the National Office. Public Information Department. PMID- 3052466 TI - Marketing your services. Strategies that work. PMID- 3052467 TI - Marketing your services. Case study 1. A private practitioner prepares for PL 99 457. PMID- 3052468 TI - Marketing your services. Case study 2. A school system wins with team players. PMID- 3052469 TI - Marketing your services. Case study 3. A hospital capitalizes on opportunities. PMID- 3052470 TI - Marketing your services. Case study 4. A clinic profits from board members' help. PMID- 3052472 TI - Marketing your services. President's page. PMID- 3052471 TI - Marketing your services. Case study 5. State association makes May a year-round event. PMID- 3052473 TI - Marketing your services. The business of grantseeking. PMID- 3052474 TI - Marketing your services. Speak with sense. PMID- 3052475 TI - Marketing your services. Quality sells. PMID- 3052477 TI - Status of communication services in nursing homes in New York State. PMID- 3052476 TI - Speech-language pathologists, audiologists, and HMOs: status & outlook. PMID- 3052478 TI - Evaluation of the requirements for the Certificates of Clinical Competence in Speech-Language Pathology and Audiology. PMID- 3052479 TI - Lack of inhibition of DNA synthesis by prostaglandin I2 in cultured intimal smooth muscle cells from rabbits. AB - The negative control system for proliferation by prostaglandin I2 (PGI2) was studied in the smooth muscle cells (SMC) cultured from the thickened intima (intimal SMC) of rabbit aortas. Indomethacin was found to enhance DNA synthesis of medial SMC but not that of intimal SMC. Exogenously added PGI2 or its stable analogue, CS-570, was observed to inhibit DNA synthesis of medial SMC enhanced by indomethacin but not that of intimal SMC. These results indicate that medial SMC are negatively controlled by endogenous PGI2 and that intimal SMC have no such negative control system for cell proliferation. This lack of negative control may be one of the mechanisms underlying the rapid growth behavior of intimal SMC as compared to medial SMC. PMID- 3052480 TI - [The onchocercal nodule]. AB - The morphology of adults worms (male and female) is characteristic particularly the content of the uterus in the female worm and the fact that the organs do not fill the body cavity. PMID- 3052481 TI - [Immunohistochemical detection of HBs, HBc and delta antigens in liver sections. Technics, value as a routine procedure according to the cycle of the viral disease and prospects for the future]. AB - The detection of hepatitis B viral antigens and the hepatitis delta viral antigen on liver section is currently facilitated by the production and marketing of good quality antisera and revelation systems. Moreover, the routine application of these techniques in histology departments has become more widespread. The detection of HBs and HBc antigens may be useful in the diagnosis of chronic hepatitis. It allows evaluation of replication of HBV (in the same way as HBe antigen and serum DNA) and the examination of a liver needle biopsy should include, in addition to analysis and classification of the lesions, tests for the presence of these antigens. The detection of the delta antigen is particularly useful for the diagnosis of acute hepatitis delta (in cases of co-infection or secondary infection). On the other hand, it is disappointing for the diagnosis of chronic hepatitis delta. PMID- 3052482 TI - [Papillary carcinoma of the thyroid: development of the histological criteria for diagnosis. Study of 29 cases and review of the literature]. AB - Papillary carcinoma (PAP) is the most frequent malignant tumour of the thyroid. PAP and follicular carcinoma of the thyroid are two biologically different tumours, without any intermediate or mixed form. Therefore the differentiation between PAP and follicular carcinoma is essential. The histological diagnosis of PAP is based upon several criteria, the most important being the papillae, the "ground glass" nuclei, and the psammoma bodies. The value of the "ground glass" nuclei in the diagnosis of PAP has been particularly emphasized in the last 10 years. The need to take into account all the diagnostic histological criteria of PAP is necessary in the definition of the different variants of PAP, particularly its follicular variant. We report 28 cases of PAP and one case of insular carcinoma, the latter probably originating from a PAP. "Ground glass" nuclei often associated with "grooved nuclei", were identified in the paraffin sections of 7 PAP. One case was classified as a follicular variant of PAP. Psammoma-bodies were seen in 6 PAP. Eleven PAP were less than 1 cm in diameter (microPAP). One microPAP presented with important lymph node metastases and blood vessels' involvement; 2 others microPAP arose in a vesicular adenoma. Six PAP were entirely encapsulated, without any lymph node metastasis or vessel invasion. Six PAP presented with lymph node metastases associated with lymphatic and/or blood vessels invasion. Lymphatic and blood vessel invasion was seen more often in association with PAP extending to the thyroid capsule. The histological classification and prognostic criteria are discussed and compared with those previously described in the literature. PMID- 3052483 TI - Treatment of paroxysmal supraventricular tachycardia in the emergency department by clinical decision analysis. AB - Vagal maneuvers terminate new onset, catheter-induced paroxysmal supraventricular tachycardia (PSVT) in up to 92% of patients. The risk and benefit of vagal maneuvers for treating PSVT in the emergency department (ED) is inadequately defined. The purpose of this study was to determine the efficacy of nonpharmacological vagal interventions in converting spontaneous episodes of PSVT in adult patients and to derive a treatment plan for such patients based on clinical decision analysis. Seventeen adult patients who presented to the ED because of PSVT were treated with carotid sinus massage, Valsalva maneuver, and head-down tilt (alone and in combination). Only three patients converted out of PSVT with vagal intervention. The remainder received verapamil, which converted 12 of the 14 patients (86%) who received the drug (one required digoxin, one required synchronized cardioversion). Vagal maneuvers are safe in young, otherwise healthy patients but problems have been documented in the literature in older patients, who have a higher likelihood of coronary and/or cerebrovascular disease. Clinical decision analysis indicates that young patients should be treated initially with vagal maneuvers but that older patients (above approximately 65 years of age) should be treated initially with verapamil. PMID- 3052484 TI - Acute recurrent appendicitis with appendicolith. AB - Appendiceal disease can be acute, acute recurrent, or chronic. Acute appendicitis is the most common form. Acute recurrent appendicitis is more common than chronic appendicitis. In children the clinical manifestations of appendicitis are variable. Patients who have an appendicolith usually develop appendicitis, often with perforation. A case is presented of 3-year follow-up of a patient with an appendicolith and acute recurrent appendicitis. The literature about appendicoliths is reviewed. In the appropriate clinical setting, a history of prior episodes of similar right lower quadrant pain does not preclude the diagnosis of appendiceal disease. Awareness of the less common forms of appendicitis is important so that appropriate treatment is not delayed. PMID- 3052485 TI - Blunt pelvic trauma. AB - Blunt pelvic trauma results in significant morbidity and mortality from associated genitourinary, neurological, vascular, and visceral damage. Diagnosis begins in the ED with the initial trauma evaluation. Proper treatment using a multidisciplinary approach and cooperation between orthopedist, urologist, trauma surgeon, and emergency physician should minimize complications. PMID- 3052486 TI - Cranial burr hole decompression in the emergency department. AB - Presently virtually all patients with acute head trauma are computed tomography (CT) scanned and transferred to a neurosurgical operating room before any surgical intervention. The time required for this, especially if the patient is transferred to another institution, may lead to a significant delay in treatment. In a patient with an expanding intracranial hematoma and evidence of brainstem compromise this delay may produce a worse outcome. Cranial burr hole placement can rapidly, safely, and accurately find and partially decompress most extracerebral intracranial hematomas. A burr hole placed rapidly before CT and transfer could prevent further damage to the brain by an expanding hematoma. The case of a child with a preterminal epidural hematoma whose outcome was excellent because of a burr hole placed in the emergency department (ED) is presented. In light of this case and a complete literature review, it is suggested that more frequent attempts to decompress intracranial hematomas in the ED may be warranted. PMID- 3052487 TI - Management of early pregnancy bleeding in the accident and emergency department. AB - Over a period of 6 months, from February to July 1987, 26,837 patients attended St Mary's A&E Department. Of these, 179 (0.7%) were complaining of bleeding in pregnancy of less than 20 weeks gestation. In May 1987 new management guidelines were introduced. These emphasised the importance of vaginal examination and the use of ultrasound to determine the viability of the pregnancy. As a result, the incidence of admission fell from 15 of 53 (28%) to 7 of 58 (12%), referral to the on-call gynaecologist from 23 of 53 (44%) to 13 of 58 (22%) and reattendance rates from 11 of 53 (15%) to 4 of 58 (4%) (all changes P less than 0.05). PMID- 3052489 TI - [The anticoagulant action of vascular endothelial cells]. PMID- 3052491 TI - [Growth in children receiving a renal transplant]. PMID- 3052490 TI - [Anaphylactic shock]. AB - The anaphylactic shock is a life-threatening reaction produced by the release of pharmacologically active substances (histamine, leukotriene...) by most cells and basophils. The release of these mediators may be immunologically mediated (anaphylactic reaction a typical immediate hypersensitivity reaction mediated by IgE) or not (anaphylactic reaction when not mediated by an antigen-antibody process). These mediators in turn specific end-organ responses in the cardio vascular system, (vasodilatation, change in inotropy, increased capillary permeability), the respiratory system (bronchospasm upper airway oedema) and the skin (urticaria). Because of its etiology (mainly drugs, contrast media and colloids) the treatment of anaphylactic or anaphylactic reactions must be prophylactic. When it occurs, its cure is based upon adrenaline and fluid loading and eventually bronchodilators. PMID- 3052492 TI - [Growth in patients with a renal transplant]. PMID- 3052488 TI - Successful transcutaneous external pacing for asystole following cardiac arrest. AB - We report a case of successful transcutaneous external pacing for out of hospital cardiac arrest causing asystole. PMID- 3052493 TI - [Recognition and diagnosis of juvenile ankylosing spondylitis: clinical analysis and comparative study of juvenile rheumatoid arthritis]. PMID- 3052494 TI - [Serratia marcescens epidemic outbreak in a neonatology service]. PMID- 3052495 TI - [Intestinal perforation caused by typhoid fever in children]. PMID- 3052496 TI - [Robinow's syndrome]. PMID- 3052497 TI - Adsorption separation processes for protein purification. PMID- 3052498 TI - Place of chromatography in the separation and purification of proteins produced from cultured cells. PMID- 3052499 TI - Disintegration of microorganisms. AB - The most common methods for the large-scale disintegration of microorganisms are high-pressure homogenization and wet milling. The most-used homogenizer is produced by Manton Gaulin and the common ball mill is the Dyno-Mill. Other manufacturers are now producing similar equipment (e.g., Rannie homogenizers and Netzsch ball mills). However, only relatively limited information on these systems has been published so far. An additional system that might become available for large-scale disintegration is microfluidization. When coming to choose optimal equipment and conditions for cell disintegration, it might be useful to consider some relevant topics that were compiled by the Retsch company for particle disruption and are presented, with modification, in Table XIII. Obviously, the importance of the above points will vary in different processes, but all of them should be considered in any biotechnological large-scale application. It is important to realize that although the mechanical disintegration methods are of general use, it is essential to define the optimal disintegration conditions for each microorganism and/or product. As a starting point, the published data for a similar product should be consulted. This should be followed by laboratory-scale experiments carried out with equipment and conditions that can be easily scaled up. PMID- 3052500 TI - Effect of curing time on marginal sealing by four dentin bonding agents. PMID- 3052501 TI - A two-year clinical study of posterior etched-porcelain resin-bonded restorations. PMID- 3052502 TI - Remineralization. A status report for the American Journal of Dentistry. Part I. PMID- 3052503 TI - Aluminum oxalate for dentin bonding. An SEM study. PMID- 3052504 TI - SEM analysis of ultrasonically debonded acid-etched metal retainers in teeth. PMID- 3052506 TI - Silver-glass ionomers. A status report for the American Journal of Dentistry. PMID- 3052505 TI - Treatment of a severely resorbed anterior vertical and horizontal maxillary ridge. PMID- 3052507 TI - Remineralization. A status report for the American Journal of Dentistry. Part II. PMID- 3052508 TI - Potential role of a liver-derived factor in mediating renal response to protein. AB - Increases in renal blood flow and glomerular filtration rate occur following the ingestion of a protein-rich meal. It has been postulated that this renal response is stimulated by some hormonal factor. Glucagon has been proposed as a probable mediating hormone, but results of recent studies argue against a direct mediating effect of glucagon. It is postulated that glomerular hyperfiltration induced by various stimuli (protein ingestion, amino acid infusion, glucagon infusion, diabetes mellitus) is associated with increased secretion by the liver of a factor that increases glomerular filtration rate. Preliminary data suggest that serotonin might play a role in mediating the postprandial increases in renal hemodynamics following protein ingestion. PMID- 3052509 TI - Protein-restricted diets and progression of renal failure. AB - Interest in dietary therapy of chronic uremia has reawakened because such therapy may slow or halt progression of renal insufficiency. The efficacies of three regimens: 0.6 g protein/kg/day; 0.3 g protein/kg/day plus essential amino acids, and 0.3 g protein/kg/day plus keto acid regimens, have been tested. Each can maintain nitrogen balance if properly administered but if dietary protein and/or the supplement are inadequate, muscle wasting will occur. Data showing that each can slow the rise in serum creatinine are presented. The problems with using serum creatinine, potential mechanisms for the effect on progression and methods for monitoring compliance are discussed. PMID- 3052510 TI - Amino acids and keto acids in the treatment of chronic renal failure. AB - Clinical trials using very low protein supply supplemented with amino acid or keto acid preparations have shown the possibility of slowing the rate of decline of renal function in patients with advanced chronic renal failure together with preservation of nutritional status. The possible mechanisms by which this effect is obtained are reviewed. The potential advantages of keto acids upon amino acids, the optimal composition of the preparations used, and the best time to start treatment in chronic uremics are discussed. Projection of recent data indicate that a prolongation of renal autonomy of about 4 years instead of a spontaneous duration of 15 months until dialysis can be expected using keto acid treatment if started when plasma creatinine reaches 500 mumol/l. PMID- 3052511 TI - Protein restriction, blood pressure and the progression of diabetic nephropathy. AB - Diabetic nephropathy is the long-term complication of diabetes responsible for the greatest increased mortality. Clinical nephropathy is characterised by a triad consisting of persistent proteinuria (total urinary protein greater than 0.5 g/24 h), rising arterial pressure and declining renal function. The role of treatment of raised blood pressure and the influence of dietary protein restriction on the established progressive phase of the disease are discussed. Subclinical elevations of urinary albumin excretion rates (greater than 30 micrograms/min; microalbuminuria), glomerular hyperfiltration and marginal elevations of arterial pressure are early markers of later clinical nephropathy which appear to respond to strict blood glucose control, blood pressure treatment and lowered dietary protein intake. Recent evidence to suggest that an inherited predisposition to raised arterial pressure may confer the susceptibility to diabetic nephropathy is presented. PMID- 3052513 TI - [Echographic evaluation of single renal cysts]. PMID- 3052512 TI - Thyroid hormone induces synthesis and accumulation of tropomyosin and myosin heavy chain in limb buds of premetamorphic tadpoles. AB - Myosin heavy chain (MHC) and tropomyosin (Tm) have been isolated from limb muscles of the North American bullfrog, Rana catesbeiana, and injected into rabbits to raise monospecific antibodies. These antibodies were used to study the localization and synthesis of myosin heavy chain and tropomyosin in the limb buds of premetamorphic (stage VI-VII) tadpoles treated with triiodothyronine (T3) to induce metamorphosis. Indirect immunofluorescence localization detects the accumulation of both MHC and Tm in the developing thigh region within 24 h of T3 treatment. During the subsequent 48 h, the accumulation of these proteins is enhanced in the thigh and progresses from thigh to the distal regions of the limb. Quantitative immunochemical determinations indicate that within 24 h of T3 treatment, synthesis of Tm and MHC are increased 23-fold and 6-fold, respectively. Following 5 days of T3 treatment, the synthetic rates of Tm and MHC are 266 and 70 times the control values, respectively. Both methods suggest that Tm is synthesized and accumulated at a greater rate than myosin heavy chain. These observations suggest that T3 promotes the differentiation of muscle in the limb buds of premetamorphic tadpoles and that limb development promoted by T3 in tadpoles is similar to that described during the embryonic development of higher vertebrates. PMID- 3052514 TI - Trends in carcinogenesis and experimental pathology. In honorem Professor Olav Hilmar Iversen 1923-1988. PMID- 3052516 TI - Modulation of growth-stimulating and antitumor activity of macrophages by BCG and cyclophosphamide. AB - Recently it has been shown that some tumours require macrophage-derived growth factors for in vitro and in vivo proliferation, as well as tumour growth stimulation by macrophages. Furthermore, it has been reported that several biological response modifiers (BRM) stimulated some growth factor production in macrophages. In the present study we tried to define whether some BRM can modulate production of macrophage-derived growth factors for stimulating tumour cells. Results obtained indicate that: 1) growth stimulating activity of macrophages may be co-expressed with antitumor activity; 2) growth stimulating activity could prevail over antitumor effects in the outcome of tumour cell/macrophage interaction in vitro; 3) Some BRM and antitumor drugs can modulate the balance between antitumor and growth stimulating activity of macrophages. It is therefore proposed that growth factors modulation assessment is necessary for new BRM characterization. PMID- 3052515 TI - Deoxycytidine, insulin and epidermal growth factor overcome the effect of two natural inhibitors of the haemopoietic system. AB - The effect of 2'-deoxycytidine on a pluripotent haemopoietic stem cell proliferation inhibitor, as assayed in the mouse spleen colony technique and on the synthetic haemoregulatory pentapeptide (acting predominantly on myeloid determined cells) as assayed in the agar colony technique, was studied. 2' deoxycytidine was able to overcome the effect of both inhibitors. The inhibition was also blocked by insulin or epidermal growth factor. Since these results are in agreement with those on other cells and other inhibitors, they may describe a more general phenomenon. It is suggested that the mode of action of the inhibitors involves, probably indirectly, interaction with the activity of ribonucleotide reductase. PMID- 3052517 TI - Human heart transplantation. Rejection risk factors. AB - The present analysis of risk factors in human cardiac transplantation is based on a review of 682 endomyocardial biopsies from 52 grafts in 51 recipients. Acute rejection was diagnosed in 149 biopsies (21.8%). The cumulative 1-year graft survival was 91.5 4%. Four out of seven patients died of irreversible rejection. An univariate analysis using the linearized rate of total rejection showed significantly higher frequency of acute rejection when donor and recipient differed for two HLA-DR antigens compared to zero or one HLA antigen disparity (p less than 0.01), as well as in patients treated with low dose steroids, Cyclosporine (CyA) and Azathioprine (p less than 0.01), compared to treatment with CyA high dose steroids. Other risk factors were graft ischemia extending 60 minutes (p less than 0.05) and patient age exceeding 40 years (p less than 0.05). A multivariate analysis using the competing risk hazard model for irreversible (= lethal) rejection was performed. The presence of two HLA-DR mismatches between donor and recipient was found to be an independent risk factor (relative risk = 8.9), and immunosuppression with CyA and high dose steroids without Azathioprine another (relative risk = 15.3). Potential risk factors such as donor and recipient sex, donor age, prior surgery and time on extracorporeal circulation were not of significant prognostic value neither in regard to rejection nor irreversible rejection. PMID- 3052518 TI - The influence of technical procedures on cell countings in the rat corneal epithelium: a statistical analysis. AB - We have performed a statistical analysis on cell counts from different areas of the rat corneal epithelium by using the number of cells per high power microscopical vision field (1250 x) as a unit. Each vision field consisted of 182 micron basal membrane. The number of vision fields varied from 29 to 39 in the horizontal and vertical diameter sections used. Only small variations were found in the number of cells from the different vision fields inside one single section. This confirm the extreme regularity of this epithelium. However, when comparing different sections, even from the same eye, the variations were much greater. Further, sections prepared for autoradiography showed a 10% reduction of the number of cells compared with the sections only stained with hematoxylin. A large proportion of the variation in the cell counts must be a result of the technical procedures, and is thus not an intrinsic property of the corneal epithelium. This conclusion may also be valid in cell counts from other epithelial structures. PMID- 3052519 TI - The rediscovery of the macrophage. PMID- 3052520 TI - Early experiences with soft intraocular implants. AB - The author recounts his early experience with implants for the posterior chamber made of silicone. While the numbers are small the visual results appear to be satisfactory. The results, however, are marred by an as yet unacceptable rate of post-operative complications. PMID- 3052521 TI - IOL prediction: an evaluation of preoperatively determined intraocular lens power accuracy. AB - A retrospective survey of 612 eyes that had undergone cataract extraction and IOL implantation was undertaken to evaluate the accuracy of ultrasound biometry combined with keratometry using the SRK regression formula, for the preoperative prediction of intraocular lens powers. A mean error of +0.35 dioptre sphere (DS) (SD +/- 0.98) was found for the series overall, with a significant (P less than 0.005) difference between the distribution of postoperative refractive errors using the S.R.K. formula for IOL prediction and the use of a standard lens of 19.5 DS. The consistency of results was tested for those patients with greater or less than normal axial length. Linear regression analysis showed no correlation between axial length and postoperative refractive error and therefore does not support the adjustment of predicted IOL powers by a factor based on axial length. Statistically significant differences were found between surgeons' results, supporting the practice of A-constant modification for individual surgeons. PMID- 3052522 TI - Who remembers the first professor of ophthalmology? AB - The cradle of modern ophthalmology was the Joseph's Academy which still stands in Vienna's ninth district. History records some famous names from the academic lineage which flourished there in the nineteenth century, but few people have heard of Joseph Barth, who became the first professor of ophthalmology in 1773. A biography of Joseph Barth is presented. PMID- 3052523 TI - The legal aspects of medical defence. AB - Defence organisations provide many varied "services" to members. Litigation involving doctors includes giving reports, giving evidence in court for or against the plaintiff or defendant, appearing as a witness at inquiries, e.g. coronial inquiries, and actions against doctors, generally alleging negligence. Patients do not seek treatment with the intent of eventually commencing proceedings. What are the catalysts for a patient commencing legal action or seeking legal advice? What should the doctor do to assist his/her defence? These questions are addressed in the article. PMID- 3052524 TI - Measurement of choroidal melanomas: a comparison of methods. AB - The prognosis for death from metastatic choroidal melanoma following enucleation has been shown to be strongly correlated with a number of risk factors of which the most important are age of onset, aggressive cell types and tumour volume. The advantages of enucleation for the treatment of choroidal melanomas are put into question by the singular lack of a parallel increase in life expectancy following this treatment, and evidence that it may promote the development of metastatic disease. Alternative forms of treatment have been introduced including observation of small and asymptomatic tumours. We are using a computer-aided system for serial measurement and statistical analysis of area and volume of choroidal melanomas. A comparison of results using our method and the conventional method of estimating volume by the product of basal area and height for 51 measurements on 15 eyes over a 6-month period showed an overestimation of volume by conventional methods which could be corrected by applying a "shape constant" determined by linear regression. Calculation of tumour growth rates is also shape-dependent, and a slowing in growth rate or even a reduction in melanoma size is possible. PMID- 3052525 TI - Complicated strabismus and adjustable sutures. AB - Clinical experience in performing over 500 adjustable strabismus operations is mentioned in this paper particularly as it relates to complicated strabismus. A maximum hang-loose recession of a rectus muscle has a limited effect, which is tabulated and the implications discussed. Aids in finding the lost medial rectus muscle are mentioned. Adjustable sutures are then exploited in managing the found "lost" medial rectus. Adjustable recession of both vertical recti in the affected eye were used in some cases of blow-out fracture to manage limitations of upward and downward gaze. Adjustable recession of both yoke medial recti are used in some unilateral superior oblique palsies where the main sequela is hypertropia in downward gaze only. A previously paralysed lateral rectus muscle, which has completely recovered function but has left the patient with a concomitant esotropia with full ductions and normal versions, responds excessively to resection. This should be taken into consideration when planning adjustable strabismus surgery in such a case. PMID- 3052526 TI - Current concepts in the management of scleritis. AB - Scleritis is an important, severely destructive, chronic inflammatory disorder affecting the eye wall. It presents a difficult management problem, often requiring high-dose systemic corticosteroid therapy or other immunosuppressive regimens to control the inflammatory response. A quantitative scleritis scoring system has been developed and its application to the assessment and management of scleral disease is discussed. This paper reviews current concepts in the management of scleral disease with emphasis on newer treatment modalities, such as pulse therapy with intravenously administered methylprednisolone or cyclophosphamide, and the use of orally administered cyclosporin. PMID- 3052527 TI - Sir Norman Gregg lecture. PMID- 3052528 TI - Clinical investigation of retinitis pigmentosa. AB - The diagnostic features and future research directions of retinitis pigmentosa were documented in this update and review of the subject. An extensive and current bibliography is provided. PMID- 3052530 TI - Successful pterygium surgery. PMID- 3052529 TI - Post-cataract astigmatism: its control and correction. AB - The factors responsible for postoperative astigmatism--wound compression and wound gape--are discussed and illustrated for a number of suture incision combinations. Variability is pronounced. Mid-limbal incisions sutured with 8-0 virgin silk had a range of 8D at four days, 8.5D at four weeks and 9D at three months. Nylon 9-0 double continuous, tied normally, had 8D at four days, 7D at four weeks and 7.5D at three months. The same suture tied tightly gave a range of 5D at four days, 7.5D at four weeks and 6D at three months. Methods of control and correction are discussed and illustrated with case histories. PMID- 3052531 TI - Non-radioactive oligonucleotide probe for detection of clinical enterotoxigenic Escherichia coli isolates of bovine origin. AB - Oligonucleotides (32 or 34 mer) corresponding to enterotoxigenic Escherichia coli STIa (ST-P) toxin gene were tailed with Bio-11-dUTP using terminal deoxynucleotidyl transferase. Plasmids from clinical E. coli isolates were prepared by modified rapid alkaline lysis procedure and dot-spotted. Biotinylated oligonucleotide probes were hybridized to detect the STIa toxin gene. The results were in agreement with that obtained by radioactive oligonucleotide probes. Of 135 clinical isolates (sampled from 6 different regions of France), only 7 (5.2%) were found to be STIa+. These 7 isolates were the only ones to be found positive for the K99 adhesive pili antigen. Both the probes were specific to the STIa toxin gene and failed to detect the closely related STIb (ST-H) toxin gene. Possibilities of their wide usage in clinical labs are discussed. PMID- 3052532 TI - On interspecies gene transfer: the case of the argF gene of Escherichia coli. PMID- 3052533 TI - Brucellosis in humans: its diagnosis and treatment. PMID- 3052534 TI - Viral diseases in developing countries. PMID- 3052535 TI - Streptococcal diseases. PMID- 3052536 TI - Comparative study of culture, direct immunofluorescent antibody test and enzyme immunoassay for the diagnosis of urogenital infections caused by Chlamydia trachomatis. AB - We studied three population groups (161 symptomatic female, 62 asymptomatic female and 74 symptomatic male) to determine the prevalence of genital tract infections caused by Chlamydia trachomatis in Kuwait. Three techniques, viz. tissue culture, direct immunofluorescent antibody test (MicroTrak) and enzyme immunoassay (Chlamydiazyme) were employed for the detection of C. trachomatis. The overall positivity based on positive tissue culture in symptomatic cases was 16.2% (17.4% in female and 13.5% in male). The two rapid methods for antigen detection were compared with tissue culture. Based on a positivity criterion of greater than 10 elementary bodies in the direct immunofluorescent antibody assay, sensitivity and specificity of the test were 66.7% and 97.2% while the positive and negative predictive values were 77.8% and 97.2% respectively. The sensitivity and specificity of the enzyme immunoassay were 61.3% and 96.7% with positive and negative predictive values of 80.8% and 94.6% respectively. There was an agreement of 92.0% between the three tests. The two rapid antigen detection tests seem to be similar and provide an alternative for diagnosing Chlamydia trachomatis in our patients. PMID- 3052537 TI - Response of Plasmodium falciparum to antimalarial drugs in vitro at Wad Medani, Sudan. PMID- 3052538 TI - A general survey of blood and tissue parasites of some rodents in Arbil province, Iraq. AB - In a survey of blood and tissue parasites from 233 mice and rats, consisting of 105 specimens of house mice (Mus musculus), 99 black rats (Rattus rattus), and 29 Norway rats (Rattus norvegicus), collected from different localities of Arbil province, northern Iraq, Trypanosoma lewisi was found in 7 (7.07%) of 99 R. rattus and in 5 (17.86%) of 29 R. norvegicus, while Trypanosoma musculi was found in 4 (3.80%) of 105 M. musculus and this is the first record of this trypanosome from rodents in Iraq. During the preparation of tissue impression smears we came across 2 specimens of Fasciola hepatica in the livers of 2 R. norvegicus and the incidence of infection was 6.89%. Examination of other organs revealed no parasites. F. hepatica which is recorded, in the present study, for the first time from rodents in Iraq, is of particular importance since it infects both humans and domestic animals. PMID- 3052539 TI - Bactericidal and opsonizing effects of human serum on Escherichia coli O7 K1. PMID- 3052540 TI - Immunoglobulin isotype response to the group-A streptococcal carbohydrate in humans. AB - We used an ELISA to determine the levels of specific anti-GAS carbohydrate IgG, IgM and IgA in 34 patients with acute rheumatic fever (ARF) with or without carditis, in 15 patients with acute glomerulonephritis (AGN) and in 18 control patients with noncomplicated GAS pharyngitis. Patients with ARF and AGN showed a significantly higher geometric mean titer as well as a higher frequency of elevated Ig of the IgG, IgM and IgA A-CHO class antibodies during the acute stage of this disease, when compared to controls. The IgM and IgA geometric means of the antibody were higher in ARF with carditis patients as compared to the non carditis or AGN patients; however, the differences were not significant. In addition a lower frequency of antibody decline was observed in ARF with carditis patients who were seen in follow-up after a 1-year period, supporting previous observations of the persistence of this antibody in patients with rheumatic heart disease. PMID- 3052541 TI - [Cell kinetics of oligodendroglioma and oligo-astrocytoma--Ki-67 PaP study]. AB - In order to determine the principal histologic features in distinguishing the "anaplastic" from the "well differentiated" oligodendroglioma and mixed glioma (oligo-astrocytoma), correlations between the growth fraction, and histologic grade and 6 histologic variables (vascular proliferation, cytologic pleomorphism, calcification, necrosis, cellularity, mitotic index) were studied in 24 patients. The growth fraction was calculated as the percentage of Ki-67-immunostained nuclei in frozen sections using Ki-67 monoclonal antibody. 6 histologic variables were checked with HE stain of paraffin-embedded tissue sections. The growth fraction was in general agreement with the histologic grade, in order of decreasing mean percentage, ranging from 11.1% (anaplastic mixed glioma) to 5.3% (anaplastic oligodendroglioma), 2.8% (isomorphic oligo.) and 1.9% (mixed, grade II). There was a significant association between the growth fraction and 3 histologic variables (vascular proliferation, cytologic pleomorphism, mitotic index) out of 6. Thus, even though there exist some gaps between these parameters in paraffin-embedded tissues and those in frozen sections, the percentage of Ki 67-immunostained nuclei is likely to be very valuable as a supplement information of histological grading and a prognostic indicator. PMID- 3052542 TI - The smallpox story: from variolation to victory. PMID- 3052543 TI - Beriberi--another Asia-Pacific connection. PMID- 3052544 TI - Tropical macrocytic anaemia: the investigations of Lucy Wills in India. PMID- 3052545 TI - The retention in dentine and composite resin materials of Bondent dentine pins. PMID- 3052546 TI - The Weaver Reading Room and Dental Museum, University of Liverpool. PMID- 3052547 TI - Fissure sealants and the preventive resin restoration on the NHS. PMID- 3052548 TI - "Vomiting during the taking of dental impressions". PMID- 3052549 TI - Partial denture design in general dental practice--10 years on. PMID- 3052550 TI - Sealant restorations (preventive resin restorations). An addition to the NHS armamentarium. PMID- 3052551 TI - Badges of the dental profession. Association of British Dental Surgery Assistants. PMID- 3052552 TI - The end of the p value? PMID- 3052553 TI - Demonstration of the ascending aorta in infective endocarditis by intravenous digital subtraction angiography. AB - Four patients with infective endocarditis were examined by digital subtraction angiography immediately before operation. In three a root abscess was suspected and the remaining patient was believed to have a false aneurysm at an infected aortic cannulation site. In all the cases digital subtraction angiography showed the structure in several projections and confirmed the presence of a cavity. Subsequent operation confirmed the site and nature of the lesions. PMID- 3052554 TI - Fetal well-being and maternal awareness. PMID- 3052555 TI - Transcutaneous electrical nerve stimulation in the management of acute postoperative pain. AB - Twenty patients undergoing decompressive lumbar laminectomy were randomly allocated, in a double-blind manner, to receive active or inactive transcutaneous electrical nerve stimulation (TENS) as part of the management of their postoperative pain. All patients received the same non-narcotic general anaesthetic. The efficacy of the TENS was assessed by using a patient-controlled analgesia system (PRODAC) which delivered morphine i.v. This system recorded the number of demands for analgesia and the total dose administered in the first 24 h. In addition, plasma morphine concentrations were measured hourly for the first 6 h and again at 24 h. There was no statistical difference between the two groups in the number of patient demands for analgesia, morphine dose or plasma morphine concentration. TENS offered no advantage over a placebo in the management of acute postoperative pain in these patients. PMID- 3052556 TI - Oral nalbuphine for the treatment of pain after dental extractions. AB - A randomized, double-blind comparison of nalbuphine 30 mg or 60 mg by mouth and dihydrocodeine 30 mg by mouth was conducted in 75 patients with moderate to severe pain after surgery for dental extractions under general anaesthesia. A significant reduction in pain intensity followed each treatment and persisted throughout the 4-h observation period after nalbuphine, but only for 3 h after dihydrocodeine was given. Reduction in pain intensity was significantly greater 2, 3 and 4 h after the use of nalbuphine 60 mg than following dihydrocodeine 30 mg, and the mean total pain intensity difference was greater following nalbuphine 60 mg than following dihydrocodeine. Nalbuphine 60 mg effectively provided complete or good pain relief in more than 50% of the patients and only three patients in this group required additional analgesia during the period of observation, compared with nine patients in each of the other groups. However, the patients who received nalbuphine 30 mg had a significantly higher mean pain intensity before treatment than those in the other groups. The side-effects encountered were those typical of opioid medication; there were no statistically significant differences between the groups. PMID- 3052557 TI - Recent advances in therapy for hypertension. PMID- 3052558 TI - If I had an attack of malaria. PMID- 3052559 TI - Lipid-lowering therapy in perspective. PMID- 3052560 TI - Urinary gonadotrophin peptide--isolation and purification, and its immunohistochemical distribution in normal and neoplastic tissues. AB - A urinary gonadotrophin peptide (UGP) was isolated and purified from semi purified human chorionic gonadotrophin (hCG), prepared from pregnancy urine. The peptide showed hCG-B subunit activity and no hCG-alpha subunit activity as demonstrated by binding studies with the relevant antibodies. It had a molecular weight significantly less than hCG-B subunit. The peptide was linked to thyroglobulin and this conjugate used to immunise rabbits and mice. A radioimmunoassay (RIA) using 125I-UGP and the rabbit antiserum (AK12) was used to monitor chromatographed urine fractions from patients with ovarian carcinoma, seminoma and hydatidiform mole. UGP was also found in the urine extract of a healthy male, but at a much lower level. In each case the UGP detected had the same molecular weight as the pregnancy preparation and appeared to be the main gonadotrophin constituent in those urine samples. Initial immunohistochemical screening of normal and neoplastic tissues with the rabbit antibody (AK12) showed reactivity with some tumours including carcinomas of the lung, ovary, cervix and breast as well as trophoblastic and germ cell tumours. Reactions with non neoplastic tissues were confined to some specialised epithelia and macrophage populations. A more comprehensive immunohistochemical study was made using a monoclonal antibody to UGP (2C2), with a monoclonal antibody to conformational hCG (INN 13) and another monoclonal antibody to free B subunit (1E5) as controls. Similar patterns of reactivity were produced by the AK12 and 2C2 antibodies in both neoplastic and non-neoplastic tissues. Additional tissues were investigated with the three monoclonal antibodies. The 2C2 antibody reacted with 93% (77/83) of tumours examined; the INN 13 antibody reacted with only the syncytiotrophoblast cells of choriocarcinoma, hydatidiform mole, placental site trophoblastic tumour, and in one case of seminoma; the 1E5 reactivity was confined to only choriocarcinoma syncytiotrophoblast cells. PMID- 3052561 TI - Estramustine binding protein and anti-proliferative effect of estramustine in human glioma cell lines. AB - Four human cell lines derived from malignant gliomas were immunohistochemically examined for their content of estramustine-binding protein (EMBP). EMBP was detected in a large amount in all glioma cells during the entire cell cycle. EMBP has previously been demonstrated to be the major receptor protein in prostatic cancers for the cytostatic drug estramustine-phosphate (EMP). EMP caused a dose dependent inhibition of exponentially growing cells by increasing the number of cells in G2/M stage of the cell cycle as monitored by flow cytofluorometry. The effect may be coupled to arrest of the glioma cells at metaphase. The presence of EMBP may suggest a selective binding and effect of EMP in glioma cells. PMID- 3052562 TI - Morphological damage induced by Escherichia coli lipopolysaccharide in cultured hepatocytes: localization and binding properties. AB - Lipopolysaccharides (LPS) from Gram-negative bacteria are considered to be the responsible agents for the induction of endotoxic shock, affecting the liver as a target organ. In this study, the cell morphology and some biochemical properties of 24 h-culture-hepatocyte monolayers treated with Escherichia coli 0111:B4 lipopolysaccharide, were observed. Cell morphology was observed by scanning electron microscopy and immunofluorescence methods. LPS interaction induced an increase in rounded cells with diminished adhesion capacity. As biochemical parameters, albumin synthesis and 2-deoxyglucose uptake were measured. LPS decreased the hexose uptake in a dose-dependent manner. Binding of (14C)LPS to cultured hepatocytes showed that LPS binds to non-specific constituents of the membrane bilayer. PMID- 3052563 TI - A morphological study of the effect of treatment with the antibiotic ceftazidime on experimental staphylococcal endocarditis and aortitis. AB - The morphological effects of antibiotic therapy with either single or repeated (8 hourly) injections of ceftazidime were examined in rabbits with experimentally induced staphylococcal endocarditis and aortitis. At 3 h after initiating treatment, many of the bacteria, irrespective of the location of the colony, showed evidence of abnormal ultrastructural changes of the cytoplasm and/or cell wall. By 8 h many degenerate lysed bacteria were present. By 24 h, in rabbits which received a single injection, bacterial colonies contained many normal and dividing bacteria. In comparison, bacterial colonies at 24 h in rabbits receiving repeated injections consisted of large masses of lysed bacteria with only a few viable appearing thick-walled 'persistent' cells. At 48 and 72 h, no viable appearing bacteria were observed although they could be isolated by culture methods. Treatment was associated with an increased inflammatory cell response at the surface of the vegetation and within the vasculature. In the later stages there was evidence of healing with endothelialization of the lesions. It would appear, therefore, that ceftazidime penetrates efficiently into the vegetations with only a short transitory phase at sub-bactericidal concentrations. The few 'persistent' bacteria appear to be protected from the host defences by the surrounding thrombus which prevents their eradication. PMID- 3052564 TI - Alpha 1-acid glycoprotein and hepatic fibrosis. AB - The relationship between alpha 1-acid glycoprotein (alpha 1-AG) and liver fibrosis was studied. Immunoperoxidase staining of liver specimens from patients with chronic hepatitis showed large amounts of alpha 1-AG to be located primarily in hepatocytes adjacent to areas of piecemeal necrosis, bridging fibrosis and fibrous septa. In patients with severe chronic active hepatitis, hepatocytes throughout the liver stained similarly to those adjacent to areas of piecemeal necrosis. In cell cultures of human embryonal lung (HEL) fibroblasts, addition to the culture medium of alpha 1-AG promoted cell growth. It is known that alpha 1 AG is also produced in the liver. It thus appears likely that alpha 1-AG is a promoter of hepatic fibrosis in chronic hepatitis. PMID- 3052565 TI - Diphencyprone in the treatment of long-standing alopecia areata. AB - Thirty-six patients with alopecia areata of 1-54 years duration entered a study of treatment with the contact allergen diphencyprone for 8 months. Following sensitization the diphencyprone was applied to one half of the scalp at weekly intervals, the other half acting as a control. Once hair growth was established on one side, the other side was treated. Seven patients did not continue treatment and one patient showed spontaneous regrowth. Of the remaining 28 patients who persisted with treatments, fourteen (50%) regrew hair on the treated side; eight (29%) had a cosmetically acceptable result with the regrowth of terminal hair over the whole scalp. No statistically significant differences were found in age or duration of alopecia between those who regrew and those who did not. We have found diphencyprone to be an effective stimulator of hair growth in patients with severe and long-standing alopecia areata. PMID- 3052566 TI - Extracellular matrix of the marrow microenvironment. PMID- 3052567 TI - Marrow transplantation following busulfan and cyclophosphamide as treatment for translocation (4;11) acute leukaemia. PMID- 3052568 TI - Upper and lower time limits in the decision to recommend marrow transplantation for patients with chronic myelogenous leukaemia. AB - Long-term survival of patients with chronic myelogenous leukaemia (CML) requires marrow transplantation from a histocompatible donor. The optimal timing of the transplant is difficult to determine because of the high peritransplant mortality of 20-35% and the existence of a group of patients who can have the disease controlled by drug treatment for prolonged periods. We have developed a mathematical model implemented with a computer program which calculates lower and upper time limits for the timing of marrow transplantation. The lower time limit for transplantation is derived from the loss of life expectancy with delay, and the upper time limit is calculated by comparing the transplant survival probability with the probability of surviving an additional year without a transplant. Thus, an objective basis is provided for bracketing the most appropriate time for transplantation. This analysis suggests that the decision to transplant can be postponed in some patients for periods longer than may generally be recommended. PMID- 3052570 TI - Occupational health aspects of the arsenic extractive industry in Britain (1868 1925). AB - A historical survey into the occupational hazards of mining and refining arsenic is presented together with the measures adopted for their control. The industry is placed in the social perspective of its time and it is suggested that this experience could be of value to those who may encounter similar problems where arsenic is extracted elsewhere in the world. PMID- 3052569 TI - Work and pregnancy. PMID- 3052572 TI - Addition of hyaluronidase to lignocaine with adrenaline for retrobulbar anaesthesia in the surgery of senile cataract. AB - A double-blind trial demonstrates the effectiveness of adding hyaluronidase to lignocaine with adrenaline in producing ocular akinesia and anaesthesia in retrobulbar nerve blocks. 92% of the blocks in which hyaluronidase was used for intracapsular cataract surgery were judged successful compared with 56% of those without added hyaluronidase (p less than 0.01). PMID- 3052573 TI - Clinical assessment of retinal elevations: a review of methods and a novel clinical technique. AB - A new clinical test for the detection of retinal elevation is described. The test, based on alterations of retinal surface light reflexes during indirect ophthalmoscopy, is extremely sensitive to very shallow detachments. The optics, the degree of sensitivity, and the limitations of the new clinical technique are examined and other clinical methods are reviewed. PMID- 3052571 TI - Effects of intensified insulin treatment on retinal vessels in diabetic patients. AB - Forty-five diabetic patients were randomly assigned to treatment with continuous subcutaneous insulin infusion (CSII), multiple injections (MI), and conventional insulin treatment (CIT). They were prospectively followed up for one year. A computerised scanning microdensitometer was applied on fundus photographs of retinal vessels, and we studied changes in calibres of the blood column (W0) and in width (Wr/W0) and intensity (Ir) of the central 'light reflex'. After six months of improved metabolic control the Ir was reduced in both MI and CSII cases compared with CIT cases (p less than 0.01), indicating haemorrheological changes in the retinas. Within these six months cotton-wool spots appeared in half the patients (n = 15) on CSII and MI, but not in CIT patients. Subjects who developed cotton-wool spots, compared with those who did not, had greater intensities of reflection and larger calibres of vessels at the start of the study (p less than 0.01). On intensifying the treatment they were characterised by a larger fall in hemoglobin A1 (p less than 0.01) and by a larger decrease in Ir on arteries (p less than 0.05) and veins (p less than 0.01). The behaviour of the retinal circulation is different in patients developing transient ischaemic lesions on intensified insulin treatment from its behaviour in those who do not. PMID- 3052574 TI - Protein structures from NMR. PMID- 3052575 TI - Absence of hepatic cytochrome P450bufI causes genetically deficient debrisoquine oxidation in man. AB - The common genetic deficiency of drug oxidation known as debrisoquine/sparteine type polymorphism was investigated with bufuralol as prototype substrate. In human liver microsomes the 1'-hydroxylation of bufuralol is catalyzed by two functionally distinct P-450 isozymes, the high-affinity/highly stereoselective P450bufI and the low-affinity/nonstereoselective P450bufII. We demonstrate that P450bufI is unique in hydroxylating bufuralol in a cumene hydroperoxide (CuOOH) mediated reaction whereas P450bufII is active only in the classical NADPH- and O2 supported monooxygenation. In microsomes of liver biopsies of in vivo phenotyped poor metabolizers of debrisoquine or sparteine, the CuOOH-mediated activity was drastically reduced. Rabbit antibodies against a rat P-450 isozyme with high bufuralol 1'-hydroxylase activity (P450db1) precipitated exclusively P450bufI type activity from solubilized microsomes. Western blotting of microsomes with these antibodies revealed a close correlation between the immunoreactive protein and CuOOH-mediated (+)-bufuralol 1'-hydroxylation. No immunoreactive protein was detected in liver microsomes of in vivo phenotyped poor metabolizers. These data provide evidence for a specific deficiency of P450bufI and are consistent with the complete or almost complete absence of this protein in the liver of poor metabolizers. PMID- 3052576 TI - Evidence for concerted kinetic oxidation of progesterone by purified rat hepatic cytochrome P-450g. AB - Purified cytochrome P-450g, a male-specific rat hepatic isozyme, was observed to metabolize progesterone to two primary metabolites (6 beta-hydroxyprogesterone and 16 alpha-hydroxyprogesterone), two secondary metabolites (6 beta,16 alpha dihydroxyprogesterone and 6-ketoprogesterone), and one tertiary metabolite (6 keto-16 alpha-hydroxyprogesterone). The Km,app for the formation of these products from progesterone was determined to be approximately 0.5 microM, while the Km,app for metabolism of 6 beta- and 16 alpha-hydroxyprogesterone was found to be 5-10 microM. The ratio of primary to secondary metabolites did not change significantly at progesterone concentrations from 6 to 150 microM, and a lag in formation of secondary metabolites was not observed in 1-min incubations. Concerted oxidation of progesterone to secondary products without the intermediate products leaving the active site was suggested by these results and confirmed by isotopic dilution experiments in which little or no dilution of metabolically formed 6 beta,16 alpha-dihydroxyprogesterone and 6-keto-16 alpha hydroxyprogesterone was observed in incubations containing a mixture of radiolabeled progesterone and unlabeled 6 beta-hydroxyprogesterone or 16 alpha hydroxyprogesterone. Incubation of 6 beta-hydroxyprogesterone with a reconstituted system in an atmosphere of 18O2 resulted in greater than 90% incorporation of 18O in the 16 alpha-position of 6 beta,16 alpha dihydroxyprogesterone but no incorporation of 18O into 6-ketoprogesterone, even though the reaction was dependent upon enzyme and O2, and not inhibited by mannitol, catalase, or superoxide dismutase. Factors which characterize the metabolism of progesterone by cytochrome P-450g in terms of active-site constraints and the catalytic competence of the enzyme in microsomes were also explored. PMID- 3052577 TI - Dihydrofolate reductase from Escherichia coli: the kinetic mechanism with NADPH and reduced acetylpyridine adenine dinucleotide phosphate as substrates. AB - Kinetic studies on the reaction catalyzed by dihydrofolate reductase from Escherichia coli have been undertaken with the aim of characterizing further the kinetic mechanism of the reaction. For this purpose, the kinetic properties of substrates were determined by measurement of (a) initial velocities over a wide range of substrate concentrations and (b) the stickiness of substrates in ternary enzyme complexes. Stickiness is defined as the rate at which a substrate reacts to give products relative to the rate at which that substrate dissociates. Stickiness was determined by varying the viscosity of reaction mixtures and the concentration of one substrate in the presence of a saturating concentration of the other substrate. The results indicate that NADPH is sticky in the enzyme NADPH-dihydrofolate complex, while dihydrofolate is much less sticky in this complex. At higher concentrations, NADPH functions as an activator through the formation of an enzyme-NADPH-tetrahydrofolate from which tetrahydrofolate is released more rapidly than from an enzyme-tetrahydrofolate complex. Higher concentrations of dihydrofolate also cause enzyme activation, and it appears that this effect is due to the ability of dihydrofolate to displace tetrahydrofolate from a binary enzyme complex through the formation of a transitory enzyme tetrahydrofolate-dihydrofolate complex. As NADPH and dihydrofolate function as activators and as NADPH behaves as a sticky substrate, the kinetic mechanism of the dihydrofolate reductase reaction with the natural substrates is steady-state random. By contrast with NADPH, reduced 3-acetylpyridine adenine dinucleotide phosphate exhibits only slight stickiness and does not function as an activator.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3052578 TI - Mechanism of the reaction catalyzed by dihydrofolate reductase from Escherichia coli: pH and deuterium isotope effects with NADPH as the variable substrate. AB - The variations with pH of the kinetic parameters and primary deuterium isotope effects for the reaction of NADPH with dihydrofolate reductase from Escherichia coli have been determined. The aims of the investigations were to elucidate the chemical mechanism of the reaction and to obtain information about the location of the rate-limiting steps. The V and V/KNADPH profiles indicate that a single ionizing group at the active center of the enzyme must be protonated for catalysis, whereas the Ki profiles show that the binding of NADPH to the free enzyme and of ATP-ribose to the enzyme-dihydrofolate complex is pH independent. From the results of deuterium isotope effects on V/KNADPH, it is concluded that NADPH behaves as a sticky substrate. It is this stickiness that raises artificially the intrinsic pK value of 6.4 for the Asp-27 residue of the enzyme dihydrofolate complex [Howell, E. E., Villafranca, J. E., Warren, M. S., Oatley, S. J., & Kraut, J. (1986) Science (Washington, D.C.) 231, 1123] to an observed value of 8.9. Thus, the binary enzyme complex is largely protonated at neutral pH. The elevation of the intrinsic pK value of 6.4 for the ternary enzyme-NADPH dihydrofolate complex to 8.5 is not due to the kinetic effects of substrates. Rather, it is the consequence of the lower, pH-independent rate of product release and the faster pH-dependent catalytic step. At neutral pH, the proportion of enzyme present as a protonated ternary enzyme-substrate complex is sufficient to keep catalysis faster than product release.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3052579 TI - Propagation of allosteric changes through the catalytic-regulatory interface of Escherichia coli aspartate transcarbamylase. AB - Each of two previously isolated strains of Escherichia coli containing a single nonsense codon within the pyrB gene was suppressed with four different nonsense suppressors. The kinetic analysis using crude extracts of these nonsense suppressed strains indicated that the mutant aspartate transcarbamylases had altered cooperativity and affinity for aspartate as judged by the substrate concentration at half of the maximal velocity. Both pyrB genes were cloned and then sequenced. In both cases, a single base change was identified which converted a glutamine GAC codon into a TAC nonsense codon. Both mutations occurred in the catalytic chain of aspartate transcarbamylase and were identified at positions 108 and 246. The glutamine at position 108 in the wild-type structure is located at the interface between the catalytic and regulatory chains and is involved in a number of interactions with backbone and side chains of the regulatory chain. The glutamine at position 246 in the wild-type structure is located in the 240s loop of the enzyme. Two additional mutant versions of aspartate transcarbamylase were created by site-directed mutagenesis to further investigate the 108-position in the structure, a glutamine to tyrosine substitution at position 108 of the catalytic chain, and an asparagine to glycine change at position 113 of the regulatory chain, a residue which interacts directly with glutamine-108 in the wild-type structure. Both mutant enzymes have reduced affinity for aspartate. However, the Tyr-108 mutant enzyme exhibits a reduced Hill coefficient while the Gly-113 enzyme exhibits an increased Hill coefficient. The response to the allosteric effectors ATP and CTP is also changed for both the mutant enzymes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3052580 TI - 31P NMR studies of enzyme-bound substrate complexes of yeast 3-phosphoglycerate kinase. 1. Effects of sulfate and pH. Mg(II) affinity at the two ATP sites. AB - 31P NMR measurements were made (at 121.5 MHz and 5 degrees C) on enzyme-bound substrate complexes of 3-phosphoglycerate kinase in order to address three questions pertaining to (i) the integrity of the enzyme-substrate complexes with Mg(II) in the presence of sulfate concentrations typical of those used for crystallization in X-ray studies, (ii) the relative affinities of Mg(II) to ATP bound at the two sites on the enzyme, and (iii) the pH behavior of the different phosphate groups in the enzyme complexes. 31P chemical shift and spin-spin coupling constant changes showed that at concentrations of 0.5 M and higher, sulfate ion interferes with Mg(II) chelation to ATP and ADP free in solution as well as in their enzyme-bound complexes. The effect on enzyme complexes is stronger for the E.MgATP complex than for the E.MgADP complex. Sulfate ion (50 mM) also causes a approximately 0.5 ppm upfield chemical shift of the 31P resonance of enzyme-bound 3-P-glycerate even in the absence of ATP or Mg(II). A quantitative estimate of the dispartate affinities of Mg(II) to ATP bound at the two sites on the enzyme was made on the basis of computer simulation of changes in the line shape of beta-P (ATP) resonance and of changes in 31P chemical shift of the corresponding gamma-P (ATP) in the E.ATP complex with increasing [Mg(II)]. The concentrations of the relevant species that contribute to these 31P NMR signals were computed by assuming independent binding at the two sites.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3052581 TI - 31P NMR studies of enzyme-bound substrate complexes of yeast 3-phosphoglycerate kinase. 2. Structure measurements using paramagnetic relaxation effects of Mn(II) and Co(II). AB - Measurements of the paramagnetic effects of two dissimilar activating paramagnetic cations, Mn(II) and Co(II), on the spin relaxation rates of the 31P nuclei in the complexes of 3-phosphoglycerate kinase with ATP, ADP, and 3-P glycerate have been used to study the structures of these enzyme-substrate complexes. All experiments were performed on enzyme-bound complexes, so that two exchanging complexes (with and without cation) contribute to the observed relaxation rate. Measurements were made at three 31P NMR frequencies, 81, 121.5, and 190.2 MHz, and as a function of temperature in the range 5-20 degrees C to determine the effect of exchange on the observed relaxation rates. Relaxation rates in E.MnADP and E.MnATP were shown to be exchange-limited, and therefore bereft of structural information, both by lack of frequency dependence and by temperature dependence with activation energies (delta E) in the range 5-8 kcal/mol. Relaxation rates for E.CoADP and E.CoATP exhibit frequency dependence and delta E values in the range 1-3 kcal/mol; i.e., these rates depend on the Co(II)-31P distances. Difficulties involved in estimating electron relaxation times in E.CoADP and E.CoATP restrict calculation of Co(II)-31P distances in these complexes to upper and lower limits. These distances were all in the range 2.7-4.1 A, appropriate for direct coordination of Co(II) to the phosphate groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3052582 TI - Penetration of the signal sequence of Escherichia coli PhoE protein into phospholipid model membranes leads to lipid-specific changes in signal peptide structure and alterations of lipid organization. AB - In order to obtain more insight in the initial steps of the process of protein translocation across membranes, biophysical investigations were undertaken on the lipid specificity and structural consequences of penetration of the PhoE signal peptide into lipid model membranes and on the conformation of the signal peptide adopted upon interaction with the lipids. When the monolayer technique and differential scanning calorimetry are used, a stronger penetration is observed for negatively charged lipids, significantly influenced by the physical state of the lipid but not by temperature or acyl chain unsaturation as such. Although the interaction is principally electrostatic, as indicated also by the strong penetration of N-terminal fragments into negatively charged lipid monolayers, the effect of ionic strength suggests an additional hydrophobic component. Most interestingly with regard to the mechanism of protein translocation, the molecular area of the peptide in the monolayer also shows lipid specificity: the area in the presence of PC is consistent with a looped helical orientation, whereas in the presence of cardiolipin a time-dependent conformational change is observed, most likely leading from a looped to a stretched orientation with the N terminus directed toward the water. This is in line also with the determined peptide-lipid stoichiometry. Preliminary 31P NMR and electron microscopy data on the interaction with lipid bilayer systems indicate loss of bilayer structure. PMID- 3052583 TI - Separation and characterization of three insulin receptor species that differ in subunit composition. AB - Partially purified human placental insulin receptor preparations give rise to three distinct insulin-binding peaks when eluted from a Mono Q high-performance liquid chromatography anion-exchange column. We analyzed the basis for this phenomenon by affinity cross-linking of insulin to each peak, followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. We find that the three insulin-binding peaks represent different molecular weight complexes with the following subunit composition: (alpha beta)2, (alpha beta)(alpha beta'), and (alpha beta')2, where beta' represents a proteolytically derived fragment of the beta subunit. This analysis of subunit composition was confirmed by silver staining of affinity-purified insulin receptor following resolution of the forms on a Mono Q column as described previously. We have characterized the three isolated insulin receptor forms with regard to ligand binding by LIGAND and Scatchard analysis. We also measured insulin-stimulatable autophosphorylation and exogenous kinase activity directed toward poly(Glu/Tyr) (4:1). The three forms of the insulin receptor exhibit similar KD's for insulin binding to the high- and low-affinity sites. The (alpha beta)2 and (alpha beta)(alpha beta') forms of the insulin receptor display superimposable curvilinear Scatchard plots. In contrast, only the intact holoreceptor (alpha beta)2 form demonstrates insulin-stimulatable autophosphorylation and exogenous kinase activity. The (alpha beta)(alpha beta') form has reduced basal kinase activity which was not increased by prior incubation with insulin. The (alpha beta')2 form lacks a kinase domain and consequently demonstrated no kinase activity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3052585 TI - Effect of zinc ions on tRNA structure: imino proton NMR spectroscopy. AB - The structure of tRNA in solution was explored by NMR spectroscopy to evaluate the effect of divalent cations, especially zinc, which has a profound effect on the chromatographic behaviour of tRNAs in certain systems. The divalent ions Mg2+ and Zn2+ have specific effects on the imino proton region of the 1H NMR spectrum of valine transfer RNA (tRNA(Val] of Escherichia coli and of phenylalanine transfer RNA (tRNA(Phe] of yeast. The dependence of the imino proton spectra of the two tRNAs was examined as a function of Zn2+ concentration. In both tRNAs the tertiary base pair (G-15).(C-48) was markedly affected by Zn2+ (shifted downfield possibly by as much as 0.4 ppm); this is the terminal base pair in the augmented dihydrouridine helix (D-helix). Base pair (U-8).(A-14) in yeast tRNA(Phe) or (s4U 8).(A-14) in tRNA1(Val), which are stacked on (G-15).(C-48), was not affected by Zn2+, except when 1-2 Mg2+ ions per tRNA were also present. Another imino proton that may be affected by Zn2+ in both tRNAs is that of the tertiary base pair (G 19).(C-46). The assignment of this resonance in yeast tRNA(Phe) is tentative since it is located in the region of highly overlapping resonances between 12.6 and 12.3 ppm. This base pair helps to anchor the D-loop to the T psi C loop.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3052584 TI - Expression and stability of c-sis mRNA in human glioblastoma cells. AB - The production of platelet-derived growth factor like (PDGF-like) material by glioblastomas may be involved in the conversion of normal cells to tumor cells. In an investigation of this problem, we have examined some of the properties of the platelet-derived growth factor B-chain mRNA (c-sis mRNA) by a sensitive and quantitative RNA-RNA solution hybridization method. In 5 out of 8 human glioblastoma cell lines, c-sis mRNA was present, and in the line with the highest level, there were approximately 4-10 molecules per cell. The half-lives of the c sis mRNA in two glioblastoma cell lines were 2.6 and 3.4 h, while in human umbilical vein endothelial (HUVE) and bladder carcinoma (T24) cells they were 1.6 and 2.5 h, respectively. Inhibiting protein synthesis produced no significant alteration of the c-sis mRNA half-lives in the glioblastoma or HUVE cells. The A U-rich sequence at the 3' end of the c-sis mRNA therefore does not appear to affect the mRNA stability in the presence of cycloheximide as it does in other transcripts. The similarity of the c-sis mRNA half-lives in normal and tumor cells suggests that regulation of stability of c-sis mRNA is not a major factor in tumorigenesis in the glioblastoma cell lines examined. PMID- 3052586 TI - 8-Ketodeoxycoformycin and 8-ketocoformycin as intermediates in the biosynthesis of 2'-deoxycoformycin and coformycin. AB - An enzyme has been isolated from cell-free extracts of Streptomyces antibioticus that can catalyze the reduction of 8-ketodeoxycoformycin (8-KetodCF) and 8 ketocoformycin (8-ketoCoF) to the naturally occurring nucleoside analogues 2' deoxycoformycin (dCF) and coformycin (CoF), respectively. The partially purified reductase requires NADPH as the cofactor and stereospecifically reduces the 8 keto group of both ketonucleoside substrates to a hydroxyl group with the R configuration at C-8. This is the same configuration of the hydroxyl group as that of the dCF and CoF isolated from S. antibioticus. The reduction proceeds at the nucleoside level, and ATP is not required. The reductase is stereospecific for the NADPH cofactor in that it transfers the pro-S but not the pro-R hydrogen from C-4 of NADPH to the 8-keto group. The apparent Km for 8-ketodCF and 8 ketoCoF were 250 and 150 microM, respectively. These in vitro results, which show that 8-ketodCF and 8-ketoCoF may be intermediates in the biosynthesis of dCF and CoF, support and extend our earlier results from in vivo studies which established that adenosine and C-1 of D-ribose are the carbon-nitrogen precursors of dCF. A possible mechanism for the formation of dCF is presented. PMID- 3052587 TI - Photoinactivation of the thiamin transport system in Saccharomyces cerevisiae with azidobenzoyl derivatives of thiamin. AB - In an attempt to obtain a potent inhibitor for thiamin transport of Saccharomyces cerivisiae three novel thiamin derivatives having an arylazido substituent in the thiazole moiety have been synthesized. The derivatives prepared were 4 azidobenzoylthiamin (ABT), 4-azidobenzoylthiamin disulfide (ABTD), and 4-azido-2 nitrobenzoylthiamin disulfide (ANBTD). Among the newly prepared photoreactive azidobenzoyl derivatives of thiamin, ANBTD showed the strongest competitive inhibition with an apparent Ki of 7.9 nM against thiamin uptake by S. cerevisiae IFO-2375. The Ki values for ABT, 4-azido-2-nitrobenzoylthiamin (ANBT), and ABTD were 187 nM, 83 nM, and 15 nM, respectively. When exposed to visible light, ANBTD inactivated in a time- and concentration-dependent manner the uptake of [14C]thiamin by yeast protoplasts as well as intact cells. Remaining activities of the thiamin uptake by the intact cells were 71.9%, 27.3%, 40.1%, and 15.0% after visible light irradiation for 15 min in the presence of 1 microM ABT, ANBT, ABTD, and ANBTD, respectively. The inactivation by ANBTD (0.05 microM) was partially prevented by previous addition of an excessive amount of thiamin (5 microM). Furthermore, it was found that ANBTD (0.5 microM) irreversibly inactivated 70.6% of the thiamin-binding activity of the membrane fraction from S. cerevisiae IFO-2375. These results suggest that ANBTD can inhibit yeast thiamin transport by photoinactivation of membrane-bound thiamin-binding protein in the plasma membrane which may be a functional component involved in the thiamin transport system of S. cerevisiae. PMID- 3052588 TI - Chemo-mechanical leak formation in human erythrocytes upon exposure to a water soluble carbodiimide followed by very mild shear stress. I. Basic characteristics of the process. AB - Human erythrocytes treated with low concentrations (1-5 mM) of the carboxyl group modifying reagent 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) lose their native deformability in parallel with extensive cross-linking of the membrane skeleton. After treatment with higher (5-40 mM) concentrations of the reagent the cells develop a hitherto undescribed property: when subjected to even very low shear stresses (resuspension after packing by centrifugation or viscometric shearing at up to 4 s-1) they become highly leaky to ions, lose their K+ with a half-time of about 5 min and subsequently undergo hemolysis. Lysis is not accompanied by cell fragmentation as occurs with mechanical hemolysis, but is colloid-osmotic, due to the formation of aqueous membrane leaks with an apparent radius of about 3 nm. Leakiness and lysis affect an increasing fraction of the cell population, in relation to (a) the concentration of EDC applied, (b) the shearing intensity, and (c) particularly, the hematocrit during shearing. The physical parameter determining the mechanical component of this 'chemo mechanical' leak formation is not predominantly the shear stress. Rather, cell cell interactions of as yet undefined nature seem to be involved. The analysis of chemo-mechanical leak formation may provide interesting insights into the influence of mechanical forces on membranes. PMID- 3052589 TI - A 2H-NMR study on the glycerol backbone of phospholipids extracted from Escherichia coli grown under high osmotic pressure: evidence for multiconformations of phosphatidylethanolamine. AB - A glycerol-requiring auxotroph was isolated from mutagenized Escherichia coli K 12 UFAts cells. This auxotroph was used for the specific deuteration of E. coli phospholipids. The cells were grown under high osmotic pressure (in the presence of 2.0% KCl). The membrane had a highly saturated fatty acid composition (76% phosphatidylethanolamine, 20% cardiolipin and 4% phosphatidylglycerol). The deuterium magnetic resonance spectra of coarse liposomes of the extracted phospholipids with perdeuterated glycerol incorporated into them were measured. To obtain well characterized information, phospholipid mixtures reconstituted from the deuterated and nondeuterated components at the same ratios as in the case of the total extract were used. On the analysis of the spectra, the following conclusions were drawn. (1) The whole polar region of cardiolipin is dynamically symmetric and quite rigid in the presence of phosphatidylethanolamine. (2) Although the quadrupole splittings of the deuterons at the C-2 and C-3 positions of the glycerol backbone were similar to each other, those at the C-1 position for phosphatidylethanolamine and cardiolipin are different, even in the same bilayer. (3) Furthermore, each C-1 deuteron of phosphatidylethanolamine gave rise to a doublet, suggesting the presence of two backbone conformations, between which there is slow exchange. (4) The polar head group of phosphatidylethanolamine interacts with cardiolipin and phosphatidylglycerol in different ways, which could be responsible for the different osmotic properties of the vesicles composed of them. PMID- 3052590 TI - Oncogenes in transgenic mice. PMID- 3052591 TI - Recognition and destruction of neoplastic cells by activated macrophages: discrimination of altered self. PMID- 3052592 TI - Cancer metastasis: tumor cell and host organ properties important in metastasis to specific secondary sites. PMID- 3052593 TI - Purification and properties of a novel recombinant human hybrid interferon, delta 4 alpha 2/alpha 1. AB - The human interferon (huIFN) delta-4 alpha 2(5-62)/alpha 1(64-166) is a genetically engineered hybrid that consists of residues 5-62 of huIFN alpha 2 and residues 64-166 of huIFN alpha 1. This variant contains four cysteine residues at positions 29, 86, 99 and 139, but does not contain the cysteine at position 1 that is characteristic of naturally occurring huIFN alpha subtypes. This novel recombinant hybrid was purified from Escherichia coli to greater than 95% homogeneity. The purification was based on ethanol extraction of a trichloroacetic acid precipitate and Matrex Gel Blue A chromatography followed by either a selective precipitation or DEAE-Sepharose chromatography. The purified protein that was treated with 2-mercaptoethanol exhibited two closely migrating bands on sodium dodecyl sulfate-polyacrylamide gel electrophoresis with apparent molecular weight values of 17,800 and 17,100, both of which exhibited antiviral activity. Electrophoresis performed without prior reduction with 2 mercaptoethanol indicated only a minor extent of intermolecular disulfide bonding. The purified protein exhibited a high specific antiviral activity of 7 x 10(7) units/mg when assayed on human fibroblast cells and, in distinction to the parental huIFN alpha 2, it also demonstrated antiviral activity on human fibroblast cells and, in distinction to the parental huIFN alpha 2, it also demonstrated antiviral activity on murine L929 cells. The level of antiproliferative activity of huIFN delta-4 alpha 2(5-62)/alpha 1(64-166) on various cell lines of different histological origin appeared to be more comparable to that of huIFN alpha 1 than huIFN alpha 2. The data suggest that huIFN delta-4 alpha 2(5-62)/alpha 1(64-166) hybrid may be a useful tool for understanding huIFN structure-function relations. PMID- 3052594 TI - Aerobic, inactive forms of Azotobacter vinelandii hydrogenase: activation kinetics and insensitivity to C2H2 inhibition. AB - Azotobacter vinelandii hydrogenase (EC class 1.12), either purified or membrane associated, was obtained aerobically in an inactive state. The kinetics of activation by treatment with a reductant (H2 or dithionite) were determined. Three distinct phases of the activation were observed. Aerobically prepared, inactive hydrogenase was insensitive to acetylene inhibition, but could be rendered acetylene-sensitive by reduction with dithionite. These findings indicate that acetylene inhibition of hydrogenase requires catalytically active enzyme. PMID- 3052596 TI - [Determination of rate constants for the antigen-antibody reaction. Kinetic characteristics of interaction of soluble and corpuscular antigen with specific antibodies]. AB - Using the previously developed procedure, the rate and equilibrium constants for the interaction of antibodies with soluble or corpuscular antigens were determined. Cow milk casein was used as a soluble antigen, while heat-inactivated Staphylococcus aureus cells--as a corpuscular antigen. The values of kinetic constants for the above reactions are close to those for various antigen- antibody pairs obtained by other investigators. PMID- 3052595 TI - Insulin regulation of glucose metabolism in HT29 colonic adenocarcinoma cells: activation of glycolysis without augmentation of glucose transport. AB - The effects of insulin on glucose transport and metabolism were examined in cultured HT29 human colonic adenocarcinoma cells. The presence of glucose transporters was verified by D-glucose displaceable [3H]cytochalasin B binding. The Kd and Bmax values from cytochalasin B binding studies were 190 +/- 30 nM and 8.4 +/- 1.4 pmol/mg protein, respectively. Glucose transport determined with 3-O methylglucose showed saturable kinetics with a Km of 5.8 +/- 0.4 mM and a Vmax of 0.047 +/- 0.003 mumol/mg protein per min at 25 degrees C. Moreover, in HT29 cells, two classes of insulin binding sites were detected in radioligand binding experiments. Although insulin failed to stimulate glucose transport, it was found to activate glycolysis in HT29 cells. Glucose consumption increased from 0.33 +/- 0.03 mumol/mg protein per h to 0.49 +/- 0.05 mumol/mg protein per h and lactate production was augmented from 0.67 +/- 0.04 mumol/mg protein per h to 0.87 +/- 0.06 mumol/mg protein per h in response to 10(-7) to 10(-5) M insulin. Insulin also enhanced mannose metabolism. Apart from these two hexoses, HT29 cells exhibited a surprisingly narrow substrate specificity. With the possible exception of glyceraldehyde, little lactate was produced from alternative substrates, including adenosine, inosine, ribose, deoxyribose, dihydroxyacetone, galactose and fructose either with or without insulin. Despite its limited utilization by the glycolytic pathway, adenosine was readily salvaged for de novo synthesis of adenine nucleotides. These findings suggest that insulin directly influences substrate utilization through the glycolytic pathway in HT29 cells without activating the glucose transport pathway. PMID- 3052597 TI - [Aggregation of membrane proteins of E. coli cells after treatment with singlet oxygen]. AB - It was found that interaction of 1O2 with bacterial membranes of E. coli cells results in covalent binding and aggregation of membrane proteins. This process was shown to be inhibited by water-soluble free radical scavengers, e. g., 1O2 quenchers (cysteine, sodium azide, histidine), of which the latter afforded the strongest inhibition. No protective effect of the fat-soluble free radical scavenger ionol (BHT) on membrane protein aggregation was observed. It was assumed that the main role in oxidative destruction of bacterial membranes (in contrast to membranes from animal sources) is ascribed to processes which are not coupled to lipid peroxidation. PMID- 3052598 TI - [Effect of the protonophore carbonylcyanide-m-chlorophenylhydrazone on the localization of secreted alkaline phosphatase in E. coli]. AB - It was shown that the total amount of synthesized alkaline phosphatase as well as the value of enzymatic activity in E. coli cells decrease in the presence of the protonophore, carbonylcyanide-m-chlorophenylhydrazone. The enzyme content in the periplasm also decreases, while the amount of the enzyme bound to the spheroplasts increases. This effect is enhanced with a rise in the protonophore concentration. An electron cytochemical analysis showed that in the presence of the protonophore, alkaline phosphatase is partly localized in the cytoplasm and on the inner surface of the cytoplasmic membrane, which is unobserved in control cells. It was assumed that carbonylcyanide-m-chlorophenylhydrazone suppresses the translocation of alkaline phosphatase across the cytoplasmic membrane and enzyme biosynthesis, on the whole. PMID- 3052599 TI - [Poly(ADP-ribosylation) of nuclear proteins in rat thymocytes]. AB - Specific antibodies to poly(ADP-ribose) were obtained and characterized. Using these antibodies, the tissue specificity of poly(ADP-ribose) modified nuclear proteins from rat thymocytes and hepatocytes was studied. The differences in the levels of poly(ADP-ribosylation) of nuclear proteins from both tissues were found to be quantitative rather than qualitative. Analysis of intranuclear distribution of poly(ADP-ribose) acceptor proteins revealed that the bulk of them is localized in the nuclear sap and matrix. A comparison of spectral properties of poly(ADP ribosylated) proteins, using specific antibodies and label incorporation from [14C]NAD showed the existence of two protein groups. Some of those were modified in a great degree but exchange poly(ADP-ribose) at a slow rate, whereas others (e.g., histones and HMG proteins) modified in a small degree exchanged poly(ADP ribose) at a much higher rate. The results obtained by different methods are discussed. PMID- 3052600 TI - [Determination of the indices of genetic similarity and diversity using programmable microcomputers]. AB - A complex of programs for microcomputer permitting to compute the similarity and diversity indices of two or few populations, herds, intrabreed types, lines and related groups of animals in automatic operation has been suggested. The absolute values of occurrence of factors and the gene frequencies in polymorphic systems of serum proteins and erythrocyte antigens of blood groups are used in calculation. PMID- 3052601 TI - Perinatal vitamin D metabolism. PMID- 3052602 TI - Ovarian follicular dynamics during the estrous cycle in heifers monitored by real time ultrasonography. AB - It is not clear whether the turnover of ovarian follicles during the estrous cycle in cattle is continuous and independent of the phase of the cycle, or whether waves of follicular growth occur at specific times of the cycle. To clarify this controversy, the pattern of growth and regression of ovarian follicles was characterized during a complete estrous cycle in ten heifers by daily ultrasonographic examinations. Follicles greater than or equal to 5 mm were measured and their relative locations within the ovary were determined in order to follow the sequential development of each individual follicle. Results indicated the presence of either two (n = 2 heifers), three (n = 7), or four (n = 1) waves of follicular growth per cycle. Each wave was characterized by the development of one large (dominant) follicle and a variable number of smaller (non-dominant) follicles. In the most common pattern observed (three waves/cycle), the first, second, and third waves started on Days 1.9 +/- 0.3, 9.4 +/- 0.5, and 16.1 +/- 0.7 (X +/- SEM), respectively. The dominant follicle in the third wave was the ovulatory follicle. The maximal size and the growth rate of the dominant follicle in the second wave were significantly lower than in the other waves, but no significant difference was observed between the first and third waves. For the two heifers that had two follicular waves/cycle, the waves started on Days 2 and 11, whereas in the remaining heifer (four waves/cycle), the waves began on Days 2, 8, 14, and 17, respectively. At 0, 1, 2, 3, and 4 days before estrus, the ovulatory follicle was the largest follicle in the ovaries in 100%, 95%, 74%, 35%, and 25% of follicular phases monitored, respectively. The relative size of the preovulatory follicle at the completion of luteolysis (progesterone less than 1 ng/ml) was negatively correlated (r = -0.90; p less than 0.0001) with the interval of time between the end of luteolysis and the luteinizing hormone surge, suggesting that the length of proestrus is determined by the size of the pre-ovulatory follicle at the beginning of proestrus. In conclusion, this study shows that the development of ovarian follicles greater than or equal to 5 mm in heifers occurs in waves and that the most common pattern is three waves per estrous cycle. PMID- 3052603 TI - Antigens recognized by monoclonal antibody to mouse acrosomal components differ in guinea pig spermatogenic cells and sperm. AB - Monoclonal antibody 1D4 reacts with a glycoconjugate antigen of the developing mouse spermatid acrosome until the terminal steps of spermiogenesis. Although 1D4 does not label the acrosome of mouse epididymal spermatozoa, it does bind to the acrosomal region of guinea pig epididymal sperm. Here we report that the antigens recognized in extracts of guinea pig spermatogenic cells and sperm are different from those detected in mouse spermatids. Three major bands of reactivity with apparent molecular weights (Mr) of 97,000-145,000, 180,000, and greater than 200,000 were detected in extracts of guinea pig sperm. Soluble antigens with the same molecular weights were released after the acrosome reaction was induced with ionophore A23187. On two-dimensional immunoblots, 1D4 recognized a microheterogeneous population of molecules in guinea pig sperm extracts. The molecules recognized by this antibody are not major Concanavalin A receptors and do not react strongly with periodic acid-Schiff's stain or periodic acid dansylhydrazine. However, comparisons of immunoblots of sperm extracts indicate that 1D4 reacted with antigens having similar molecular weights to glycoconjugates recognized by antibodies J1 and C6. Immunofluorescent labeling of guinea pig germ cells showed that 1D4 reacted only with the acrosomes of developing spermatids and sperm, and occasionally with the juxtanuclear region of spermatocytes. In general, staining was associated with the periphery of the acrosome and not with the acrosomal granule. Immunoblots of extracts of guinea pig spermatocytes, round spermatids, condensing spermatids, and sperm demonstrated that the antigens change during germ cell differentiation. Thus, 1D4 can be used as a marker of the developing acrosome for studies of the synthesis, structure, and assembly of the sperm organelle. PMID- 3052604 TI - From methodology to ethics and from ethics to methodology. AB - This editorial criticizes certain ethical and scientific deficiencies, the statistical manipulation of patients' rights and the misuse of the Social Security budget in human experimentation and therapeutics. PMID- 3052605 TI - Medical ethics in Japan. PMID- 3052606 TI - Arthritis and drug therapy in the hypertension clinic. AB - Non-gouty arthritis is quite common in hypertensive populations and there have been suggestions that drugs used to treat hypertension may be the cause. This possibility has been investigated in a case-control study. We identified 127 hypertensive patients with arthritis other than gout and matched them for age and sex to 254 hypertensive controls who did not have arthritis. Use of antihypertensive drugs was similar in cases and controls for all patients, and for those with undiagnosed arthritis. There was no significant excess use of any type of antihypertensive drug by cases with arthritis. These findings do not support the hypothesis that drug treatment for hypertension commonly causes arthritis. It is more likely that hypertension and arthritis occur together frequently because non-steroidal anti-inflammatory drugs raise blood pressure, and because both problems are common in middle-age. PMID- 3052607 TI - Role of light toxicity in the developing retinal vasculature. PMID- 3052608 TI - Spindle cells and retinopathy of prematurity: interpretations and predictions. PMID- 3052609 TI - Vitamin E and retinopathy of prematurity: the clinical investigator's perspective on antioxidant therapy: side effects and balancing risks and benefits. PMID- 3052610 TI - Cryotherapy: indications, methods, and current status. PMID- 3052611 TI - Development of the blind infant and child with retinopathy of prematurity: the physician's role in intervention. PMID- 3052612 TI - Perinatal intracranial hemorrhage and retinopathy of prematurity: currently nonpreventable complications of premature birth? AB - There is now considerable evidence that the control of cerebral circulation in sick small infants is sometimes seriously deranged and subject to wide fluctuations during neonatal care. Our current preoccupation with looking at oxygen administration and peripheral arterial oxygen tensions as the only cause of ROP is not likely to be productive. Until we know more about the causes of cerebral blood flow fluctuation, IVH, asphyxia, and their effects on the retinal circulation, we are not apt to make much progress. There is a strong association between retinopathy and IVH in premature infants. With our current knowledge, we are unable to prevent or to treat either IVH or ROP. The public should be better informed that both of these conditions are usually unavoidable complications of premature birth. PMID- 3052613 TI - Respiratory physiology, oxygen therapy, and monitoring: report of a clinical trial of constant monitoring. PMID- 3052614 TI - Growth factors: soluble mediators of wound repair and ocular fibrosis. PMID- 3052615 TI - Selected conditions with ectodermal dysplasia. PMID- 3052616 TI - Selected ectodermal dysplasias. PMID- 3052618 TI - Dental and oral abnormalities in selected ectodermal dysplasia syndromes. AB - Only a brief review of the dental and oral abnormalities in a few ectodermal dysplasia syndromes has been presented. Obviously, careful evaluation of the dentitions of patients suspected to have these disorders will add to our knowledge and assist in diagnosis of this heterogeneous group of genetic diseases. PMID- 3052617 TI - Structural abnormalities of the epidermally derived appendages in skin from patients with ectodermal dysplasia: insight into developmental errors. PMID- 3052619 TI - Dental treatment for patients with ectodermal dysplasias. PMID- 3052620 TI - Psychological aspects of hypohidrotic ectodermal dysplasia. AB - There is little information on the psychologic aspects of HED. In particular, there is insufficient evidence to support or dispute the claim that the intelligence of persons with HED is dissimilar to that of the general population. Impressions of frequent mental retardation may arise from patients suffering brain damage following high fever and from our biases against the physically unattractive. A small number of adults with HED have been reported to hold a relatively wide range of jobs, and an even smaller number have been reported with educational and occupational achievement that is predictive of bright average to superior intellectual functioning, but even less evidence is available here. Chronic illness and handicaps exert a toll on the patient and the family. There appears to be greater maladjustment among the families of chronically ill children, but this differs according to the type of illness, and we do not know how this affects the HED child and his/her family. Still, the HED family is exposed to some unique stresses: having always to take into account the ambient temperature regardless of their planned activity; responding to cruel stares and remarks when out in public; and dealing with the problem of broken, lost, or merely publicly discovered dentures. On the other hand, some children apparently benefit from having a sib with a disability. Families react in a variety of ways to a disability, and their reaction may interact with the child's temperament to affect emotional development for better or worse. However, the existence of HED does not insure either normal or inadequate emotional adjustment, and parents should be advised of this. As difficult as it may be, this may also be the time for the pediatrician to say "I don't know." Parents can be told that each child, not just those with HED, should be provided a warm and supportive environment congruent with his/her unique style, in order to facilitate emotional development. They should also be advised of the probable emotional benefits of early prosthetic care with wigs and dentures. When this is either not possible or not sufficient, professional guidance should be sought. Psychotherapy or counseling can be obtained for the child, a parent, or for the entire family. While some familiarity with HED may be helpful, it is not essential, and is far less important than finding a competent professional with whom the family is comfortable. As far as obtaining the information we need, a series of systematic investigations appears to be called for.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3052621 TI - Pathogenetic analysis of certain developmental and genetic ectodermal defects. AB - It is postulated that, in the nevoid basal cell carcinoma syndrome, independent mosaic pleiotropic action of the mutant gene on morphogenesis and histogenesis produces primary malformations of midline and nonmidline structures and dysplasias more or less highly predisposed to cancer development. However, in focal dermal hypoplasia and the Bartsocas-Papas syndrome, it is highly tempting to postulate that embryonic dysplasias, ie, breakdown or necrosis of ectoderm, especially in the region of the Ekdodermring, are responsible for the production of many of the congenital anomalies seen in these patients, and that these anomalies more likely represent sequences rather than primary malformations. The sequences in the type 1 fetal epidermal dysplasias probably represent mucosal breakdown, producing various upper gut atresias and an epidermal "disease" with loss of epidermis and many severe secondary consequences. Polyhydramnios, micrognathia, "arthrogryposis," reduced fetal growth, and short umbilical cord are other consequences of fetal hypokinesia due to stiff skin in the type 2 fetal epidermal dysplasias, with severe muscle involvement in the Hutterite-Mennonite type possibly responsible for additional fetal hypokinesia. Thus it seems likely that embryonic and fetal dysplasias can now be held responsible, directly and indirectly, for a fascinating variety of human congenital anomalies. PMID- 3052622 TI - Desmopressin: a nontransfusional form of treatment for congenital and acquired bleeding disorders. AB - Desmopressin (1-deamino-8-D-arginine vasopressin, abbreviated DDAVP) is a synthetic analogue of the antidiuretic hormone L-arginine vasopressin. Because it can raise circulating levels of factor VIII coagulant activity (FVIII) and von Willebrand factor and shorten the prolonged bleeding time, DDAVP is established as a nontransfusional form of treatment for mild and moderate hemophilia and von Willebrand disease. Recently, DDAVP has also been purported to be useful for shortening the prolonged skin bleeding times that occur with uremia, cirrhosis, and platelet dysfunctions of various etiologies. Finally, there is evidence from controlled clinical trials that DDAVP can reduce blood loss and transfusion requirements for hemostatically normal individuals undergoing spinal fusion surgery and for patients undergoing cardiopulmonary bypass surgery. The purpose of this report is to review the therapeutic applications of DDAVP in congenital and acquired bleeding disorders and to discuss areas in which further basic and clinical research is needed. PMID- 3052623 TI - Platelet glycoprotein IIb/IIIa complex in cultured cells. Localization in focal adhesion sites in spreading HEL cells. AB - A panel of mouse monoclonal antibodies (MoAbs) was raised that react with platelet glycoproteins (GP) IIb or IIIa. On immunofluorescence, the MoAbs reacted with 30% to 40% of the human erythroleukemia (HEL) cells. When the HEL cells were induced to spread on fibronectin in the presence of 12-O-tetradecanoylphorbol-13 acetate (TPA), the MoAbs reacted with the focal adhesion sites. Only some of the GPIIIa MoAbs reacted with cells other than platelets, megakaryocytes, or HEL cells, and these showed focal adhesion sites in cultured human endothelial cells, fibroblasts, and epithelial cells from normal kidney tubules. They did not react, however, with transformed fibroblasts, fibrosarcoma cells, cultured cells from hypernephromas, or cultured human amnion epithelial cells. The results suggest that the platelet-type GPIIb/IIIa complex is only expressed in cells showing an ability to define megakaryoblastic differentiation. Localization of the GPIIb/IIIa complex at the induced focal adhesion sites in HEL cells and localization of the GPIIIa-like molecules in other cells suggest their direct role in the adhesion process and in the actomyocin organization of adherent cells. PMID- 3052624 TI - Anti-K562 cell monoclonal antibody RTF8X recognizes tumor-associated antigen of erythroid lineage. AB - A new anti-K562 cell monoclonal antibody, RTF8X, a cytotoxic IgM, recognized a surface antigen on erythroblasts from patients with erythroleukemia and polycythemia vera. RTF8X, which is highly specific to K562 cells, did not react with the other 14 hematopoietic cell lines and the seven nonhematopoietic cell lines. RTF8X antigen was not detected in normal peripheral blood, but was found in less than 1% of normal marrow cells. RTF8X did not inhibit in vitro colony formation of CFU-E and BFU-E in a complement-dependent cytotoxicity assay. Cell sorting analysis showed that, morphologically, the RTF8X-positive marrow cells from the patients and normal volunteers contained more than 60% erythroblasts and that CFU-E and BFU-E were not demonstrated in cells with RTF8X antigen. Enzyme treatment suggested that RTF8X antigen was a sialoglycolipid. These results indicate that RTF8X may recognize the surface antigen found increasingly in association with tumors of erythroid lineage. RTF8X should be useful for studies of erythroid differentiation and proliferation in patients. PMID- 3052625 TI - Allogeneic bone marrow transplantation for acute nonlymphocytic leukemia in first remission. AB - Seventy-three patients with acute nonlymphocytic leukemia in first complete remission (CR) have received allogeneic bone marrow transplantation (BMT) with non-T-lymphocyte-depleted marrow obtained from matched sibling donors. The first 36 patients received a preparative regimen consisting of cyclophosphamide, 60 mg/kg/d (days -6 and -5), and 750 cGy single-dose total-body irradiation (TBI) (day -1). Subsequently, 37 patients received cyclophosphamide 60 mg/kg/d (days -6 and -5), and 165 cGy fractionated TBI administered twice daily for a total dose of 1,320 cGy (days -4, -3, -2, and -1). Survivors have been followed from 9 to 124 months (median, 40 months). The 61% (95% confidence interval [CI], 45% to 77%) projected disease-free survival (DFS) of 41 children less than 18 years old does not differ significantly from the 62% (95% CI, 49% to 73%) projected DFS of 32 adults at 84 months (P = .89). Similarly, the 15% (95% CI, 1% to 29%) projected relapse rate seen in children does not differ from the 9% (95% CI, 0% to 21%) seen in adults (P = .69). Multivariate Cox regression analysis of presenting features demonstrates that a presenting WBC count greater than 20,000/m3 is associated with decreased DFS (P = .01). When compared with other French-American-British (FAB) subtypes, presentation with FAB M4 or M5 morphology is significantly associated with relapse in multivariate analysis (P = .014). Other presenting features such as preparation with single-dose or fractionated TBI, interval from diagnosis to CR or CR to BMT, donor or recipient sex, and donor or recipient cytomegalovirus serology do not correlate independently with either DFS or relapse. When included in the stepwise multivariate analysis of presenting patient features, two posttransplant events, development of grades 2 to 4 acute graft-v-host disease (GVHD) (P less than .03) and development of interstitial pneumonitis (P less than .001), also correlate independently with poor DFS. Allogeneic BMT provides equivalent, prolonged DFS in both children and young adults when performed in first CR and should be considered the therapy of choice for all first CR patients under 45 years of age with a suitable donor. Continued efforts to prevent and treat acute GVHD and pneumonitis as well as efforts designed to prevent relapse in patients presenting with FAB M4 and M5 morphology should further improve outcome. PMID- 3052626 TI - Fractionated total body irradiation and high-dose VP 16-213 followed by allogeneic bone marrow transplantation in advanced leukemias. AB - Thirty-eight patients (median age, 21 years) with acute nonlymphoblastic leukemia (ANLL) (17 patients), acute lymphoblastic leukemia/lymphoma (ALL) (18 patients), chronic myelogenous leukemia (two patients), and refractory anemia received allogeneic bone marrow transplants from HLA-identical sibling donors or a one antigen-mismatched brother (one patient) after a preparatory regimen consisting of fractionated total body irradiation and high-dose VP 16-213 (60 to 70 mg/kg body weight). Of the 33 patients with acute leukemia who received grafts from HLA identical donors, three patients with ANLL received transplants in first remission and one patient with standard-risk ALL received a graft while in second remission. All other patients were in more advanced stages of their disease or exhibited other high-risk features. At the time of analysis, 20 of the 33 patients were alive, with 19 of them remaining in continued complete remission for 6 to 35 months (median, 18 months). The 3-year actuarial disease-free survival rate of 56.6% +/- 9.7% (SE) and the actuarial relapse rate of 11.9% +/- 6.8% (SE) demonstrate that the combination of fractionated total body irradiation and high-dose VP 16 is an effective mode of therapy in patients with advanced leukemias. Preliminary experience cautions against the use of VP 16 instead of cyclophosphamide in any clinical situation carrying an increased risk of graft rejection because the immunosuppressive potency of VP 16 is largely untested but may be inferior to that of cyclophosphamide. PMID- 3052627 TI - Comparison of cyclophosphamide, cytarabine, and etoposide as immunosuppressive agents before allogeneic bone marrow transplantation. AB - Etoposide and cytarabine have been shown to exert high antileukemic activity and are currently under study as preparatory agents before allogeneic bone marrow transplantation. However, data concerning their engraftment-promoting potency are scarce. Therefore, we tested these agents in LEW rats receiving a myeloablative dose of busulfan followed by transfer of F1 (CAP X LEW) marrow, which is unable to induce a graft-v-host reaction (GVHR). Since busulfan by itself has only minor immunosuppressive potency, graft rejection ensues unless etoposide, cytarabine, or cyclophosphamide provide additional immunosuppression to facilitate durable engraftment. Before allogeneic bone marrow transplantation in humans, 120 mg/kg of cyclophosphamide, 60 mg/kg of etoposide, or 900 mg/kg of cytarabine are the standard doses given in conjunction with total body irradiation. Seventy-five percent of these doses administered in addition to busulfan resulted in rejection rates of 75% for cytarabine and 58% for etoposide, respectively, whereas no rejections were observed with cyclophosphamide. These data indicate that etoposide and cytarabine are inferior to cyclophosphamide in their rejection preventing potential. Using either of these agents as substitutes for cyclophosphamide before allogeneic bone marrow transplantation may increase the risk of graft rejection in HLA-mismatched bone marrow transplantation and, in case of HLA identity, if T-depleted marrow is administered. PMID- 3052628 TI - Flow-cytometric detection of terminal deoxynucleotidyl transferase and other intracellular antigens in combination with membrane antigens in acute lymphatic leukemias. AB - Development of a new fixation procedure allowed flow-cytometric analysis of nuclear and other intracellular antigens in acute lymphatic leukemia (ALL). A short fixation of the cells with buffered formaldehyde acetone (BFA) rendered the cell membrane permeable, allowing the monoclonal antibodies (MoAbs) to penetrate the cell. Through this method, a rapid analysis of intracellular antigens, specific for acute lymphatic leukemia [such as terminal deoxynucleotidyl transferase (TdT), immunoglobulin M (IgM) heavy chain, and antigens recognized by the CD22 or CD3 MoAbs) was performed by flow cytometry. The surface antigens remained intact after this fixation procedure, enabling simultaneous detection of membrane and intracellular antigens. The binding of biotinylated antibodies against several B- and T-lymphoid membrane antigens was detected with streptavidin-phycoerythrin (red fluorescence), whereas the intracellular antigens were stained with FITC-labeled polyclonal antibodies, or indirectly with FITC labeled goat anti-mouse IgG (green fluorescence). Through this combination of markers, minor cell populations can be detected and a rapid and quantitative immunodiagnosis can be performed. PMID- 3052629 TI - Purging of acute myeloid leukemic cells by ether lipids and hyperthermia. AB - Ether lipids (EL) and hyperthermia have been shown to possess a relatively selective cytotoxicity to leukemic cells. In this study, the combined effects of EL (ET-18-OCH3, ET-16-NHCOCH3, or BM 41.440) and hyperthermia on the growth of hematopoietic progenitors, myeloid leukemic cell lines, and leukemic cells obtained from patients with acute myeloid leukemia (AML) were examined to determine if this combination resulted in a greater selective killing of leukemic cells than that achieved by either EL or heat alone. When the cells were treated simultaneously with EL (50 micrograms/mL) and hyperthermia (42 degrees C) for one hour, the killing of leukemic cell line cells was enhanced considerably. Among the three EL, however, the combination of ET-18-OCH3 and heat seemed to be the most cytotoxic to leukemic cell line cells with no effect on the growth of hematopoietic progenitors. An increase in the duration of treatment with ET-18 OCH3 to four hours with heat added during the last hour resulted in a further reduction of leukemic cell line cells while sparing 50% of hematopoietic progenitors after cryopreservation. The combined treatment with ET-18-OCH3 and heat also inhibited the growth of leukemic progenitors obtained from AML patients by 97% to 100%. These data indicate that the combined treatment with EL and hyperthermia might offer an efficient means to eliminate myeloid leukemic cells in vitro. PMID- 3052630 TI - Identification of risk groups for development of central nervous system leukemia in adults with acute lymphocytic leukemia. AB - The risk of development of CNS leukemia was investigated in 153 adults with acute lymphocytic leukemia (ALL) who received systemic combination chemotherapy without CNS prophylaxis. Overall, 31 patients (20%) developed CNS leukemia after a median of 6 months of therapy; the estimated 1-year incidence of CNS leukemia was 21% (SE, 3.9%). Characteristics significantly associated with CNS involvement included the presence of elevated hemoglobin creatinine, alkaline phosphatase, fibrinogen, and lactic dehydrogenase levels; B-cell leukemia; and high leukemic cell proliferative activity. Multivariate analysis identified lactic dehydrogenase levels of greater than or equal to 600 U/L and greater than or equal to 14% of cells in the S + G2M compartment to have independent additive poor prognostic significance. Patients were categorized into different risk groups for CNS leukemia with 1-year incidences ranging from 4% to 55%. While related to a high occurrence of CNS leukemia at diagnosis (33%) and subsequently (100%), the low incidence of B-cell disease excluded it from the multivariate analysis. The use of systemic chemotherapy containing multiple agents with good CNS penetration and in high doses (VAD regimen) in 90 patients was associated with a trend for lower CNS leukemia at 1 year (15% v 31%), especially in the low risk category. We propose to develop future therapies for adults with ALL that include risk-oriented CNS prophylactic approaches. PMID- 3052631 TI - Interleukin-6 (B-cell stimulatory factor 2)-dependent growth of a Lennert's lymphoma-derived T-cell line (KT-3). AB - A T lymphoma cell line (KT-3) established from a patient with Lennert's lymphoma showed macrophage-dependent growth. Macrophage-derived factors were able to replace the macrophage functions. Experiments using a variety of cytokines demonstrated that KT-3 proliferated in response to recombinant interleukin-2 (rIL 2), rIL-4, or rIL-6 but did not proliferate in response to rIL-1 alpha, rIL-1 beta, rIL-3, recombinant granulocyte colony-stimulating factor (rG-CSF), rGM-CSF, recombinant interferon-alpha (rIFN-alpha), rIFN-gamma, recombinant tumor necrosis factor (rTNF-alpha), or native IFN-beta. Polyclonal rabbit anti-IL-6 antibody almost completely neutralized the activities of macrophage-derived factors or IL 6 but not IL-2 or IL-4. Scatchard plot analysis demonstrated that KT-3 cells indeed express IL-6 receptors. The results indicate that the macrophage-derived factor that supports the growth of KT-3 is IL-6 and suggest that macrophage derived IL-6 may play an important role in the histopathogenesis of Lennert's lymphoma. PMID- 3052632 TI - Does the chemical instability of aspartyl and asparaginyl residues in proteins contribute to erythrocyte aging? The role of protein carboxyl methylation reactions. AB - As erythrocytes age in the circulation, their proteins are subjected to a wide variety of spontaneous reactions that lead to the formation of covalent derivatives. In this article, we concentrate on nonenzymatic reactions at aspartyl and asparaginyl residues, both of which are especially vulnerable targets on the protein. These residues can be altered by a combination of deamidation, isomerization, and racemization reactions that form D- and L aspartyl and D- and L-isoaspartyl residues. We present evidence that two of these modified residues are targets for an enzymatic methyl esterification reaction, and that methylation may represent the means by which cells respond to this type of protein damage. The metabolic fate of the methyl ester is unclear, but in vitro model studies with peptides and proteins suggest that this methylation can lead to the partial repair of the altered protein and can mitigate the loss of protein function. PMID- 3052633 TI - Do natural autoantibodies play an important role in the elimination of senescent or damaged red blood cells? AB - Natural autoantibodies (NAAbs) are an important constituent of circulating immunoglobins. Some demonstrate polyspecific reactivity and high idiotypic connectivity. For example, they are reactive with DNA and cytoskeleton protein; others exhibit restricted binding specificity. Among several hypotheses, we have explored the possibility that NAAbs constitute a physiologic system in the organism that participates in the elimination of dead tissues and cellular debris by opsonization and phagocytosis. PMID- 3052634 TI - Factors that accelerate or retard red blood cell senescence. AB - This article explores information concerning alterations in the time of age related red blood cell (rbc) death (rbc senescence) in experimental animals and humans. Those factors that accelerate or retard the mean time for senescent death [the mean potential rbc life-span, (T)] are discussed; specifically excluded are conditions in which rbc survival is shortened due to an increase in the rate of age-independent [random hemolysis, (k)]rbc death. The factors prolonging senescence are reduction in metabolic rate through hibernation, reduced environmental temperature, hypophysectomy and thyroidectomy, and splenectomy. In general, these processes prolong rbc senescence by about 10%-15% in the models studied to date. The failure of splenectomy to prolong rbc senescence to any physiologically meaningful extent casts serious doubt on the concept that splenic processes are a major factor in the senescence process. Rbcs made under conditions of increased erythropoiesis and/or increased metabolic rate show acceleration of senescence. Thus, rbcs of animals treated with thyroxine show a 15% acceleration of senescence. "Stress reticulocytes" and normal full-term human newborn rbc may show up to 25% reduction in (T). The maximum acceleration seen to date is 50%-90%, as seen in the rbcs of the fetal and newborn rat. rbc senescence is not accelerated in rats with splenomegaly and increased rates of random hemolysis, again casting strong doubts on the spleen's ability to alter rbc senescence by progressively modifying the rbc during successive passages through that organ. It is postulated that rbc senescence is mainly a function of the red cell's initial endowment, particularly in the dynamic ability of that cell (and its membrane) to adapt to cumulative stresses that exist during its circulation through the body. PMID- 3052635 TI - The relationship of red cell enzymes to red cell life-span. AB - Red cells are replaced before they become senescent. It is probable that red cell destruction is controlled by a biologic clock, essentially independent of metabolism, rather than by metabolic failure. The appearance of neo-antigens on the external surface of the red cell could be this "clock." PMID- 3052636 TI - The aging of the red blood cell. A multifactor process. AB - Red blood cell (rbc) senescence is associated with loss of surface sialic acid, which is the principal carrier of surface negative charge and determines the electrokinetic behavior of old rbcs. Loss of sialic acid in an old rbc is demonstrated in its decreased electric mobility and lower negative charge density, determined topographically with cationic particle labeling. Surface sialic acid determines also the mutual attraction--repulsion forces, as demonstrated in enhanced aggluinability with cationic molecules, lectins, and blood group antibodies. Loss of sialic acid accompanies ATP-depletion in vitro; thus, a T-antigen site is unmasked. Macrophages have specific receptors to the site as to newly exposed galactose and N-acetyl galactosamine sugars. Furthermore, the involvement of complement molecules in the recognition of old RBCs by macrophages has been shown. This is possibly due to loss of sialic acid or at least a regrouping--relocation of surface anionic sites due to cell shape changes from discocytes to crenated forms, which accompany both in vivo and in vitro rbc aging. In turn, shape changes are apparently controlled by the cytoskeletal network underlying the rbc membrane, which undergoes structural alteration with physiologic aging in changing the dimensions of oligomeric spectrin and the thickness of the spectrin-actin cytoskeletal assembly. PMID- 3052637 TI - Altered enzyme function and premature sequestration of erythrocytes in aged individuals. AB - Experimentation performed to determine the parameters of the life-span of the erythrocyte in hosts of various ages have determined that, in aged individuals, the rate of turnover of cells is considerably increased over that observed in young individuals. These observations are based on studies in humans, rats, mice, and rabbits in which either in situ 59Fe labelling or age-density gradient separation were used. The mechanisms for the recognition of the effete red cell in the aged host and the nature of the membrane alterations that bring about the premature sequestration are not fully understood. However, it has been consistently observed that the red cells of aged individuals have higher levels of IgG bound to their membranes than do young cells, with the most dense cells having the highest levels of immunoglobulin. Studies of most enzymes, particularly those involved in protection against oxidative damage have shown reduced activity as a function of both cell and donor age. Evidence of enzyme damage has been observed even in the youngest circulating red blood cells of old individuals. This fact leads us to hypothesize that the erythrocyte of the aged individual as it differentiates and is released from the bone marrow is less functional and partially damaged. The erythrocytes of both old and young individuals age in the circulation, accumulating subtle alterations that are recognized by the immune and/or reticuloendothelial systems and lead to sequestration. The cells of the elderly individual accumulate a greater degree of damage due to their initially reduced capacity to protect themselves from environmental stress. These alterations eventually bring them to their early sequestration. PMID- 3052639 TI - Removal of bacteria from blood by charcoal hemoperfusion. AB - E. coli bacteria were successfully removed from contaminated RBC/plasma by using a special matrix of micro-encapsulated albumin activated charcoal (ACAC). Efficacy of removing the bacteria was directly related to the amount of time the contaminated blood was in contact with the charcoal. The data indicated that the bacteria adhered to the ACAC, but that the charcoal was not bactericidal. PMID- 3052638 TI - Red blood cell substitutes: microencapsulated hemoglobin and cross-linked hemoglobin including pyridoxylated polyhemoglobin & conjugated hemoglobin. PMID- 3052641 TI - Perfluorochemicals as oxygen transport vehicles. AB - Perfluorochemical liquids (PFC's) are good solvents for gases and as such, have significant relevance to the field of biology. Approximately 40 ml of oxygen will dissolve in 100 ml of most PFC's, and carbon dioxide is at least twice as soluble. This makes these compounds attractive as vehicles for these and other gases in-vivo and in-vitro. Although many have low viscosities and surface tensions, it is necessary to emulsify them for intravenous use and organ perfusion. The emulsion particles must be very fine, usually about 150 nanometers in diameter, and stable in the presence of blood and other biological fluids. Presently the two most used emulsifying agents are Pluronic F68 and phospholipids. Emulsions made with the former are now known to cause complement activation with accompanying transient decreases in circulating leukocytes and oxygen tension. This phenomenon is reversible and usually not severe. Phospholipid-stabilized PFC emulsions do not activate complement and should therefore, be advantageous. Areas of current research utilizing PFC emulsions are in tumor therapy, by increasing the oxygenation of otherwise hypoxic cells, in ischemic heart treatment, and in alleviating the effects of cerebral ischemia. Many other applications of PFC's are being pursued, and even more research activity in this field is likely. PMID- 3052640 TI - Trends in exploitation of packed red blood cells. AB - The following trends aim to a more efficient exploitation of packed red blood cells (PRBC): 1. Improvement of the operative distribution of PRBCs for transfusions before expiration. 2. Prolongation of the expiration time by monitoring the biochemical and physical processes during banking. Maintenance of native hemoglobin and restoration or substitution of substances involved in transport of energy and of oxygen are of utmost importance. Enzymic conversion of RBCs of blood group A, B to 0 is not supposed to leave laboratory scale soon. While cryo-conservation of RBCs with glycerine is known, freeze-drying of PRBCs remains a speculation. 3. Use of PRBCs after expiration as a raw material for products applicable in medicine and biochemistry. Stroma-free hemoglobin variants (SFH) are known as effective infusable oxygen carriers in experimental animal models. However, there is little convincing evidence on the metabolism and innocuity of SFH variants in human organism. Therefore, systemic infusion of SFH solutions is not yet acceptable to clinicians even in emergency situations. On the other hand, a broader use of SFH and its variants is anticipated and regarded as prospective in organ perfusion, cardioplegy and transplantation as well as in analytical biochemistry. PMID- 3052642 TI - Long term liver preservation using artificial blood substitutes. AB - Twenty-four hour hypothermic perfusion preservation of the liver was aimed using an artificial blood substitutes. A canine liver was harvested and preserved using originally designed perfusates. In group 1, Perfluorotributylamine solution (Oxypherol solution) was used. In group 2, (Pyridoxylated hemoglobin) (Polyoxyethylene) conjugate solution (Stabilized Hemoglobin, PHP-1). In group 3 and in group 4, modified Stabilized hemoglobin, PHP-3 and PHP-4 was used. After the preservation, each liver was transplanted orthotopically. Postoperative graft function was estimated the following parameter, such as bile excretion, consciousness level, activated clotting time, and survival rate. In group 1 and group 2, improvement of such factors were not fully observed. On the contrary, in group 3 and 4, complete recovery of the function was seen and in group 4, longer survival was obtained. PHP solution was considered suitable perfusate for liver preservation. PMID- 3052643 TI - Isolated heart preservation for 24 hours: is O2 important? PMID- 3052644 TI - Recent studies on perfluorochemical (PFC) emulsion as an oxygen carrier in Japan. AB - In summary, we have discussed PFC emulsions as oxygen carriers, and concluded as follows: PFC emulsions which are clinically usable at present, such as Fluosol DA, have some negative points. For instance, there is the requirement of relatively high FiO2 level, short retention time in the blood stream, slow excretion rate from the organs, limited dosage for infusion, and so on. However, in the clinical cases of moderate acute anemia, the consumed oxygen in the PFC phase was nearly 30% of that in the hemoglobin phase in the administration of only 1,000 ml of Fluosol-DA. It strongly suggests that PFC emulsion can carry a considerable amount of oxygen to the tissues and play an important role in improving the tissue hypoxia. Moreover, we surgeons wish to say that there is a great difference for the surgeons' mental state during a surgical operation between the cases where no blood transfusion is allowed for a religious or other reason, and the cases where we may use Fluosol-DA depending on the patients' condition. In the former cases, surgeons will be under stress such that he or she might cause the patients to lose blood excessively. On the other hand, when the surgeon has something in reserve, that is, when he or she may use Fluosol-DA if needed, a good result will usually be obtained in the operation, even without actually using Fluosol-DA. One might be led to say that a key point for the success of operations is the surgeon's mental condition, freedom from anxiety.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3052645 TI - An overview of perfluoroctylbromide--application as a synthetic oxygen carrier and imaging agent for X-ray, ultrasound and nuclear magnetic resonance. PMID- 3052646 TI - Design, synthesis and evaluation of fluorocarbons and surfactants for in vivo applications. New perfluoroalkylated polyhydroxylated surfactants. AB - The progress achieved since the advent of Fluosol-DA is summarized in this paper, with special focus on the synthesis and evaluation of more adequate, reliable, industrially feasible fluorocarbons, structure/property relationships--molecular weight being recognized as the pre-eminent determining factor of both the fluorocarbon's excretion rate and the emulsion's stability-, the preparation of significantly more concentrated, more efficacious emulsions, etc. The key to further progress and better mastery of the emulsions' characteristics, especially in relation to increased shelf-life, prolonged i.v. persistence, and versatility, now lies in the development of new surfactants, better adapted to this objective. New families of well defined, monodisperse perfluoroalkylated polyhydroxylated surfactants derived from sugars and related compounds have been synthesized and fully characterized. Preliminary evaluation of their surface-active properties, emulsion stabilizing capacity and biocompatibility are reported. PMID- 3052647 TI - The safety and efficacy of perfluorochemical emulsions as blood substitutes. PMID- 3052648 TI - Discussion and considerations for the excretion mechanism of perfluorochemical emulsion. AB - It has been assumed that a mononuclear phagocyte system is related to the excretion of PFC emulsions: PFC particles are phagocytized by blood monocytes to be expelled through the lung alveoli. This monocyte-related mechanism may well explain excretion at an early stage when PFC particles are abundant in the blood stream. It does not, however, fully explain the manner by which PFC cells are released from the RES cells into the blood stream and into the adipose tissue. To explain this, the following mechanism has been proposed and discussed based on some experimental results. PFC emulsion particles taken up by the RES organs, are stripped at their surfactant layers in the cells and move across the cell membranes to the blood vessels and into other tissues such as adipose, at a rate that depends on the lipophilicity of the PFC's. In the blood stream, PFCs are delivered by lipoproteins to the lung and excreted into the expired air. Pharmacokinetical analysis with a compartmental model for the excretion also supported this proposed mechanism. PMID- 3052649 TI - Perfluorochemical emulsions as adjuncts to radiotherapy. PMID- 3052650 TI - The use of perfluorochemical emulsion in the study of pulmonary microvascular permeability. AB - The perfluorochemical exchange transfused rat was used to examine whether the interaction of plasma proteins with the luminal surface of endothelial cells effects the permeability of pulmonary capillaries. Ultrastructural immunocytochemical techniques were used to localize albumin and IgG. The results indicated that the presence of plasma proteins localized solely within the glycocalyx was sufficient to render the underlying pulmonary capillary endothelium as impermeable to intravenously injected native ferritin (pI 4.7) as that of nonexchange transfused control rats. In further experiments it was observed that the removal of circulating plasma proteins by exchange transfusion resulted in the unmasking of anionic sites on the glycocalyx. This was determined by morphometry by counting the number of intravenously injected cationized ferritin particles (pI 8.5) bound to the luminal surface of the endothelium. There was no statistically significant difference in the amount of binding to the thick and thin sides of the capillaries. These observations support the fiber matrix model of capillary permeability as formulated by Curry and Michel. PMID- 3052651 TI - Blood substitutes and the cardiovascular system. PMID- 3052652 TI - Command hallucinations and criminality: a clinical quandary. AB - Clinical literature on the role of command hallucinations in producing antisocial behavior is sparse and fragmented. This article reviews exploratory models of auditory hallucinations and the prevalence of command hallucinations in clinical and forensic settings. In addition, clinical guidelines are offered for assessing the authenticity of command hallucinations and their relevance to criminal behavior within the context of forensic evaluations. PMID- 3052653 TI - Methodological issues in behavioral immunology research with humans. AB - This paper summarizes important methodological issues that are particularly relevant for behavioral immunology research with humans. The assessment of such salient parameters as nutrition, drug/alcohol use, physical activity, and health are discussed. In addition, a number of logistical issues are addressed. PMID- 3052654 TI - Monoclonal antibodies for progesterone receptor assays and polymorphism studies in breast cancer. Comparison with radioligand assays. AB - Progesterone receptors (PR) from human breast tumors were assayed by a new method using monoclonal antibodies immobilized on beads (PR-EIA, Abbott Laboratories). EIA results were compared to those obtained with the dextran-coated charcoal method using a tritiated ligand (ORG 2058). The precision and reproducibility of the EIA method were studied over a 3-month period: intra-assay coefficients of variation were less than 6% for the range of the assay (between 5-250 fmol/ml), and inter-assay coefficients of variation calculated on 13 consecutive standard curves were less than 10%, except for standard 0 (33%). PR assays performed on 78 tumors both by EIA and radioligand (RLA) were compared. The linear regression obtained was: PR-EIA = 0.81 RLA+1 fmol/mg protein (r = 0.88). For reproducibility studies, cytosols were assayed twice during a period ranging from 1 week-3 months, both by EIA and RLA. The linear regression obtained between the second assay (B) and the first assay (A) was: B = 0.98 A + 11 fmol/mg protein for RLA (r = 0.98), and B = 0.99 A-7 fmol/mg protein for EIA (r = 0.98). To study the effect of KCl on PR-EIA, 26 tumors were homogenized in 0.4 M KCl Tris buffer and assayed both by EIA and RLA. A good correlation was obtained between the 2 methods, but higher values were obtained with PR-EIA (P = 1.6) in comparison with RLA. The addition of KCl to the cytosol showed that KCl had no effect on EIA results, but significantly lowered RLA results. To study the effect of KCl on progesterone receptor isoforms, cytosols were analyzed by chromatography on TSK G3000 SW columns, and the presence of PR was detected in each fraction by both EIA and RLA. In the absence of KCl, only the oligomeric form of PR was observed; however with both techniques, detection of this form different from tumor to tumor, emphasizing the inter-tumoral molecular heterogeneity of PR. After PR isoform dissociation by KCl (0.4 M) and chromatographic analysis of the forms obtained, monoclonal antibodies detected PR molecular forms different from those observed by radioligand; furthermore, chromatographic patterns obtained were different from one tumor to another and confirmed the inter-tumoral molecular heterogeneity of the progesterone receptor. PMID- 3052655 TI - [Diagnosis of cancers. Role of sociopsychological factors]. AB - Early diagnosis of cancer is desirable for improving prognosis and rendering treatment more effective. It depends on the type of cancer and its site in particular. Another factor to be considered is the physician, and his/her knowledge and aptitude to manage a diagnostic problem. It depends specially on the time taken by the patient to seek medical advice. The amount of time taken depends on the patients' psycho-social parameters. Patients who have been poorly educated, who are from a low social class, who are alone or who have a fear of cancer are likely to delay consultation after appearance of symptoms. All these factors are difficult to alter. Physicians should be fully aware of these factors, and be able to recognize them in patients. Good personal doctor-patient relations and subsequent quick medical management appear to be more decisive than mass screening in reducing time required for diagnosis. PMID- 3052656 TI - [Clinical pharmacokinetics of nitrosoureas]. AB - The pharmacokinetic characteristics of chloroethylnitrosourea anti-cancer agents are based on their high chemical reactivity and subsequent rapid breakdown in the plasma; and on a widespread tissue distribution due to the molecules' lipophilic properties. These physico-chemical properties explain the main problems that one may face during pharmacokinetic studies (assay method, isolation of high reactive intermediates) and that are related to the typical pharmacokinetic profile of these agents: rapid gastro-intestinal absorption, short half-life, widespread pulmonary and renal distribution, passage through the blood-brain barrier with levels within a therapeutic range, lipoprotein fixation, mainly hepatic and pulmonary metabolism, renal excretion. A drug interaction with phenobarbital has been described in the rat. The rational use of these compounds in cancer chemotherapy requires the optimisation of assay methods and a better understanding of their transformation pathways as well as their mechanism of action. PMID- 3052657 TI - Cell culture systems are more sensitive than Saccharomyces cervisiae tests for assessing the toxicity of aquatic pollutants. PMID- 3052658 TI - Brain acetylcholinesterase activity recovery following acute methyl parathion intoxication in two feral rodent species: comparison to laboratory rodents. PMID- 3052659 TI - The role of antilymphocyte globulin in the treatment of chronic acquired bone marrow failure. AB - Antilymphocyte globulin is an immunoglobulin preparation prepared from heterologous serum after the animal (horse or rabbit) has been immunised with human lymphocytes, obtained from the thymus (antithymocyte globulin, ATG) or thoracic duct (antilymphocyte globulin, ALG). The rationale for the use of ALG in the treatment of chronic acquired marrow failure is based on its immunosuppressive activity and the fact that a proportion of cases of bone marrow failure, whether affecting single or multiple haemopoietic cell lines are due to immune-mediated suppression of haemopoiesis. In addition, in vitro studies have shown that ALG also has an immunostimulatory effect on lymphokine and haemopoietic growth factor production, and may therefore directly stimulate haemopoietic progenitor cells. ALG has been used for the treatment of aplastic anaemia and acquired chronic marrow failure affecting single cell lines namely pure red cell aplasia (PRCA), amegakaryocytic thrombocytopenia and chronic neutropenia due to immune inhibition of granulopoiesis ('acquired white cell aplasia'). ALG is used for treatment of non-severe aplastic anaemia (NSAA) and in those cases of severe aplastic anaemia (SAA) where allogeneic transplantation is not possible or is not indicated. Treatment with ALG results in 75% long term survival for NSAA and 40-50% for SAA although there is a very severe subgroup of SAA defined by peripheral blood neutrophils of less than 0.2 x 10(9)/l who rarely benefit from ALG therapy. For those patients who do not respond a second course of ALG can be given later using ALG from a different animal source.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3052660 TI - Management of refractory idiopathic thrombocytopenia. AB - In refractory thrombocytopenia one should first evaluate whether the therapeutic approach has more risks to the patient than no treatment at all. The patient may remain relatively asymptomatic and the only incommoding circumstances be cosmetic. Coincidental medical problems such as hypertension and peptic ulceration are particularly worrying in the patient with ITP. Very occasionally a cerebral haemorrhage may occur without any obvious predisposing cause. In most instances, however, refractory thrombocytopenia is a benign condition which rarely directly causes the death of the patient. Since many of the remedies advocated for this problem have significant side effects these must always be carefully balanced and fully discussed with the patient before their introduction. PMID- 3052661 TI - Clinical use of intravenous immunoglobulin in blood disorders. AB - The protective effect of passively administered immunoglobulin has been known for many years. Although its accepted use over the last 40 years has been in the prevention and treatment of infection its widest and, arguably, most important application has been in the prevention of Rhesus haemolytic disease of the newborn. However, the development over the last decade of a number of safe and effective immunoglobulin preparations for intravenous use has allowed its more widespread utilisation in clinically appropriate settings. It is now widely accepted that intravenous immunoglobulin (IVIgG) is indicated, on a repeat basis, as treatment of choice as replacement therapy for patients with primary antibody deficiency. It may also be effective, in the non-immune deficient patient, in the management of certain bacterial as well as viral infections. This is particularly the case in the pre-term neonate but may have an application in the infected intensive care patient. The benefit in these patient groups has led to a re evaluation of the use of immunoglobulin replacement therapy in the severe secondary antibody deficiency states associated with such haematological conditions as multiple myeloma and chronic lymphocytic leukaemia. These are currently the subject of randomised, controlled studies. The observation that the use of IV IgG may be associated with effects other than passive transfer of antibody were first made by Imbach in Switzerland. While treating children with immunodeficiency he noticed that two with a coincident immune thrombocytopenia had a rapid platelet increment in close association with the immunoglobulin infusion. He subsequently confirmed that this was a rapid and predictable form of therapy in childhood idiopathic thrombocytopenic purpura (ITP) and that in some children it appeared to affect the natural history of the disease. Since these initial observations the response has been confirmed in both children and adults and extended to other immune-based haematological disorders. PMID- 3052662 TI - Deoxyuridine suppression: biochemical basis and diagnostic applications. AB - The deoxyuridine (dU) suppression test evolved out of investigations into the biochemical basis of the megaloblastic changes seen in vitamin B12 and folate deficiency. Although the abnormality in dU suppression which occurs in vitamin B12- or folate-deficient states is assumed to reflect impaired methylation of deoxyuridylate, there is still no direct demonstration that this is so. Furthermore, there is evidence that reactions other than the methylation of deoxyuridylate are involved in the phenomenon of dU suppression. Nevertheless, in clinical practice abnormal dU suppression serves as a sensitive index of the presence of megaloblastosis due to vitamin B12 or folate deficiency. dU suppression is also abnormal in a number of conditions other than vitamin B12 or folate deficiency, but its overall specificity in detecting tissue dysfunction due to these two deficiency states is considerably higher than that of the serum vitamin B12 or red cell folate levels. Consequently, the test enables us simply and rapidly to define those patients in whom macrocytosis is unrelated to a deficiency of vitamin B12 or folate. For these reasons, the dU suppression test has been adopted by several laboratories across the world for investigating patients with (a) possible vitamin B12 or folate deficiency, (b) macrocytosis, and (c) megaloblastic erythropoiesis. Since the dU suppression test is abnormal in transcobalamin II deficiency and in some congenital disorders of vitamin B12 and folate metabolism, it is very useful in the investigation of obscure anaemias in infancy and childhood. In addition, it has contributed to our understanding of the mechanisms underlying the myelotoxicity of certain drugs, and particularly of nitrous oxide. PMID- 3052663 TI - Cyclic neutropenia: a clinical review. AB - Cyclic neutropenia is a benign, hematologic disorder characterized by recurrent episodes of severe neutropenia at 21 day intervals. There are associated cyclical variations in other blood cells. Patients with this disease have malaise, stomatitis, cervical lymphadenopathy and fever during the recurrent neutropenic periods. The exact cause of cyclic neutropenia is unknown. About one third of human cases appear to be inherited in an autosomal dominant pattern. In the other cases, the disease appears to arise spontaneously with symptoms usually beginning in infancy or early childhood. In adult patients, the disease may be acquired and occur in association with a clonal proliferation of large granular lymphocytes. Clinical studies in man and investigations in grey collie dogs, which have a very similar disease, strongly suggest that cyclic neutropenia is due to an abnormality in the regulation of early hematopoietic precursor cells. Therapy for cyclic neutropenia involves local and symptomatic therapy for the recurrent mouth ulcers and pharyngitis, and antibiotics for episodes of sinusitis, pneumonia, peritonitis, or bacteremia. Therapy with glucocorticosteroids, androgens, and plasmapheresis has been efficacious in a few adult patients, but no therapy has been proven to alter the cycling of blood counts in children. Despite their repetitive illnesses, patients with cyclic neutropenia grow and develop normally. With the help of attentive physicians and dentists, their quality of life and life expectancy are good. Current research on hematopoietic growth factors offers promise of new approaches to therapy. PMID- 3052664 TI - Classification of acute leukaemias. AB - Acute leukaemias have traditionally been classified according to the nature of the predominating cells as judged by cytomorphology and cytochemistry. A codification of this classification into L1-L3 subdivisions for lymphoblastic cases (ALL) and M1-M7 for myeloid cases (AML), proposed by the FAB group of haematologists, has been used extensively in the past decade. Some criticisms of this codification are presented and other approaches to classification are discussed. Among these are included the potential importance of multiple lineage expression, measurements of cell differentiation using cytochemical criteria, the relevance of isoenzyme biochemistry of leukaemic cell populations, their surface antigenic differences as demonstrated by the use of monoclonal antibodies and the growing contribution of cytogenetics. The development and complexity of combined classifications which attempt to incorporate most of these features is illustrated. A simplified classification into broad groups is proposed, ALL being divided by surface antigenic and cytogenetic criteria and AML by morphology and cytochemistry. These groups may be further elaborated as appropriate. PMID- 3052665 TI - The preleukemic syndrome (hematopoietic dysplasia). AB - It has been recognized for many decades that epithelial dysplasia can represent an early histological sign of epithelial neoplasia. So it is with hematopoietic tissue wherein dysplasia of bone marrow cells can be an early sign of impending acute myeloid leukemia. Although this 'preleukemic syndrome' of hematopoietic dysplasia can often be identified well in advance of the classic histological signs of acute leukemia, a wide variety of basic studies on bone marrow cells, from patients and from experimental animals with induced preleukemia, clearly indicate that the preleukemic marrow cells are members of a fully established neoplastic clone. Consequently, it is likely that the preleukemic syndrome is merely acute leukemia diagnosed earlier than usual and which, in some patients, can be very slowly progressive, and in others may not progress at all. This article reviews the evidence in support of the notion that the preleukemic syndrome is an 'early leukemia', places the preleukemic syndrome in the context of a larger group of myelodysplastic disorders, reviews the laboratory studies of value for both diagnosis and for use in the assessment of prognosis, and summarizes the therapeutic options available for the management of patients with this disorder. PMID- 3052666 TI - Infectious complications of blood transfusion. AB - Numerous infectious diseases are transmissible by blood, with AIDS and hepatitis being the predominant concerns today. Less in the limelight, but nonetheless blood transmissible, are cytomegalovirus infection, malaria, babesiosis, and hepatitis B. A major controversy with respect to non-A non-B hepatitis relates to the use of 'surrogate' testing of donors for ALT and hepatitis B core antibody. Transfusion-associated AIDS has been markedly reduced as a risk, due to blood donor antibody screening implemented in March 1985. However, other retroviruses such as HTLV-1, HTLV-II and HIV-II pose additional concerns regarding the safety of the blood supply, and decisions will be forthcoming regarding testing of donated blood for antibody to these viruses. PMID- 3052667 TI - Genes on the X and Y chromosomes controlling sex. PMID- 3052669 TI - ABC of eyes. The glaucomas. PMID- 3052668 TI - Randomised controlled trial of general practitioner intervention in patients with excessive alcohol consumption. AB - OBJECTIVE: To determine effectiveness of advice from general practitioners to heavy drinkers to reduce their excessive alcohol consumption (35 U or more a week for men, 21 U or more for women). DESIGN: Randomised, controlled double blind trial over 12 months with interim assessment at six months. SETTING: Group practices (n = 47; list size averaging 10,000) recruited from Medical Research Council's general practice research framework, mostly in rural or small urban settings. PATIENTS: Patients recruited after questionnaire survey. Of total of 2571 (61.2%) of 4203 patients invited for interview who attended, 909 (35.4%) stated that in past seven days they had drunk above the limits set for study and had not received advice; they were randomised to control and treatment groups. INTERVENTIONS: Patients in treatment group were interviewed by general practitioner (who had had a training session) and received advice and information about how to reduce consumption and also given a drinking diary. END POINT: Study aimed at detecting a reduction in proportion of men with excessive alcohol consumption of 30% in treatment group and 20% in control group (for women 40% and 20%, respectively) with a power of 90% at 5% level of significance. In addition, corroborative measures such as estimation of gamma-glutamyltransferase activity were included. MEASUREMENTS AND MAIN RESULTS: At one year a mean reduction in consumption of alcohol of 18.2 (SE 1.5) U/week had occurred in treated men compared with a reduction of 8.1 (1.6) U/week in controls (p less than 0.001). The proportion of men with excessive consumption at interview had dropped by 43.7% in the treatment group compared with 25.5% in controls (p less than 0.001). A mean reduction in weekly consumption of 11.5 (1.6) U occurred in treated women compared with 6.3 (2.0) U in controls (p less than 0.05), with proportionate reductions of excessive drinkers in treatment and control groups of 47.7% and 29.2% respectively. Reduction in consumption increased significantly with number of general practitioner interventions. At one year the mean value for gamma glutamyltransferase activity had dropped significantly more in treated men (-2.4 (0.9)IU/l) than in controls (+1.1(1.0)IU/l; t = 2.7, p less than 0.01). Reduction in gamma-glutamyltransferase activity tended to increase with number of intervention sessions in men. Changes in gamma-glutamyltransferase activity in women and changes in other indicators in both sexes did not differ significantly between treatment and control groups. CONCLUSIONS: If the results of this study were applied to the United Kingdom intervention by general practitioners could each year reduce to moderate levels the alcohol consumption of some 250000 men and 67500 women who currently drink to excess. General practitioners and other members of the primary health care team should therefore be encouraged to include counselling about alcohol consumption in their preventive activities. PMID- 3052670 TI - [Academic eulogy for Professor Paul Bordet, titular member and former president]. PMID- 3052671 TI - [Anomeric specificity of glucose metabolism]. PMID- 3052672 TI - [Purification, characterization and immunological study of 2 protein antigens of the BCG strain Mycobacterium bovis]. PMID- 3052673 TI - [Academic eulogy for Professor R. Dubois-Manne, honorary member]. PMID- 3052674 TI - Proposition for a new continuous suturing technique for microvascular anastomosis: a comparative study. AB - The authors describe a suturing technique for end-to-end anastomosis in small vessels, which involves the use of continuous suturing without resorting to rotation of the approximator. The method is described in detail. PMID- 3052675 TI - Split skin grafting using topical local anaesthesia (EMLA): a comparison with infiltrated anaesthesia. AB - The analgesic efficacy of EMLA cream was compared with that produced by infiltration of lignocaine solution when used to provide anaesthesia for cutting of skin grafts. The study was performed as an open parallel group comparison in 80 patients. Pain felt during administration of the anaesthetic and during cutting of the graft was assessed using visual analogue and verbal rating scales. During graft cutting, the anaesthesia produced by EMLA was at least as effective as infiltration. On administration, infiltration produced varying amounts of pain in all patients, but in contrast EMLA produced no discomfort. In view of this lack of discomfort and the consequent greater freedom afforded regarding the area of donor site anaesthetised, EMLA can be considered the treatment of choice when skin grafts are harvested under local anaesthetic. PMID- 3052676 TI - Pain-free cutting of split skin grafts by application of a percutaneous local anaesthetic cream. AB - The use of a novel percutaneous anaesthetic preparation for the pain-free cutting of split skin grafts has been assessed in a series of 80 patients, age range 12 to 96 years. The technique was completely successful in 80% of these cases, with complete analgesia established within one hour of initial application. Duration of anaesthesia was such that pain-free cutting of the skin was possible up to 5 hours after initial application of the preparation. Failures of the technique were largely attributed to a loss of anaesthetic potency in the preparation, application of an inadequate amount to the site or patient anxiety. The use of the percutaneous anaesthetic preparation was found to offer considerable advantages over conventional infiltration anaesthesia for this type of surgery. Several additional surgical applications of percutaneous anaesthesia are also considered. PMID- 3052677 TI - History of BAPS (British Association of Plastic Surgeons) PMID- 3052678 TI - Effect of adenosine on the formation of prostacyclin in the rabbit isolated heart. AB - 1. The effect of adenosine on cardiac biosynthesis of prostacyclin (PGI2) was investigated. Rabbit hearts were perfused according to Langendorff at controlled pressure (with or without theophylline), or at controlled flow. The content of 6 keto-prostaglandin1 alpha (6-keto-PGF1 alpha, metabolite of PGI2) in the coronary effluent under basal conditions and during infusion of adenosine was determined using a highly specific radioimmunoassay. 2. In other experiments, rings of rabbit aorta were incubated with or without adenosine and the production of 6 keto-PGF1 alpha was analysed as above. 3. Administration of adenosine (10 micron) to hearts perfused at controlled pressure increased the coronary flow by up to 38%. The peak concentration of 6-keto-PGF1 alpha in the effluent exceeded the control by 177% (P less than 0.01), and the total efflux of 6-keto-PGF1 alpha exceeded the control by 179% (P less than 0.001). Theophylline (50 micron) reduced these effects of adenosine by 23%, 43% and 51%, respectively, without influencing the uptake of adenosine into the heart. 4. When adenosine (1-10 micron) was administered to hearts perfused at controlled flow, a dose-dependent decrease in the perfusion pressure, by 27% and 44% respectively, was observed. In parallel, the resulting increase in 6-keto-PGF1 alpha efflux was considerably lower (49% (P less than 0.05) and 43% (NS), respectively). A similar decrease in perfusion pressure induced in the absence of adenosine decreased the efflux of 6 keto-PGF1 alpha, by 15% (P less than 0.01) and 32% (P less than 0.001), respectively. 5. Addition of adenosine (1-10 microM) to incubates of rabbit aortic rings did not significantly affect the concentration of 6-keto-PGF1 alpha in the incubation medium in comparison with control. 6. We conclude that adenosine stimulates rabbit heart PGI2 formation, mainly by an action related to the vasodilator effect of the nucleoside. PMID- 3052679 TI - Effects of leukotriene D4 on the mechanical and electrical properties of guinea pig isolated trachealis. AB - 1. The effects of leukotriene D4 (LTD4) on mechanical and electrical activity were examined in guinea-pig isolated trachealis muscle and compared with two other bronchoconstrictors, methacholine and potassium chloride (KCl). 2. LTD4 elicited concentration-dependent increases in tension in trachealis muscle which were slower in time course than responses induced by either methacholine or KCl. The maximum response to LTD4 was approximately 85% of the methacholine maximum. 3. At a concentration close to the EC50 for tension changes, LTD4 had no significant effect on either transmembrane potential or slow wave activity recorded in single trachealis cells. 4. At a concentration close to the EC90 for tension changes, LTD4 caused significant membrane depolarization, transiently reduced the amplitude and increased the frequency of slow wave discharge and ultimately abolished slow wave discharge. LTD4-induced depolarization was less marked, and developed more slowly, than that evoked by either methacholine or KCl. 5. These results show that LTD4 can elicit substantial increases in tension without altering transmembrane potential and are consistent with the view that LTD4 initiates contraction mainly through potential-independent mechanisms. However, at high concentrations the depolarization evoked by LTD4 allows the possibility that potential-dependent mechanisms may contribute to the spasm. PMID- 3052680 TI - Muscarinic inhibition of [3H]-noradrenaline release on rabbit iris in vitro: effects of stimulation conditions on intrinsic activity of methacholine and pilocarpine. AB - 1. Rabbit isolated irides were loaded with [3H]-noradrenaline and superfused with Tyrode solution. The inhibition by the muscarinic agonists (+/-)-methacholine and pilocarpine of the [3H]-noradrenaline overflow into the superfusate evoked by field stimulation (pulses of 1 ms duration, 75 mA) was measured as an index of activation of presynaptic muscarinic receptors. 2. The fractional rate of release per pulse during the first stimulation period (S1) was low with 360 pulses at 3 Hz, intermediate with 360 pulses at 10 Hz and high with 1200 pulses at 10 Hz. Upon repetitive stimulation (7 periods at 20 min intervals), the fractional rates of release per pulse during S7 no longer differed, suggesting a 'long-term' regulation of [3H]-noradrenaline release depending on the stimulation conditions. 3. The evoked [3H]-noradrenaline overflow was depressed by (+/-)-methacholine in a concentration-dependent manner. The EC50 ranged from 0.29 to 0.42 microM. Methacholine nearly abolished the transmitter release evoked at 3 Hz but reduced that induced at 10 Hz by only 50%. Under the latter condition the methacholine concentration-inhibition curve was bell-shaped and no muscarinic inhibition was observed in the presence of methacholine 30 microM. After washout of methacholine the evoked [3H]-noradrenaline release was temporarily enhanced. 4. Atropine 0.1 microM enhanced the [3H]-noradrenaline overflow (evoked by stimulation with 360 or 1200 pulses at 10 Hz), probably antagonizing a presynaptic inhibition by endogenous acetylcholine. The inhibition by methacholine was competitively antagonized by atropine 0.1 microM (apparent -log KB = 8.5-9.0). 5. Depending on the concentration, pilocarpine reduced the [3H]-noradrenaline overflow evoked by 360 pulses at 3 Hz up to 63%. However, at 10 Hz stimulation frequency the compound was inactive as an agonist but competitively antagonized the presynaptic inhibition induced by methacholine. The KB under the latter condition (0.95 microM) was very close to the EC50 value determined at 3 Hz (0.85 microM). 6. The results demonstrate a muscarinic inhibition of noradrenaline release from the rabbit isolated iris. The activation by pilocarpine of the presynaptic receptors provides an alternative explanation for the miosis induced in the rabbit in vivo, which might be the result of a decreased sympathetic tone in the iris dilator muscle. PMID- 3052682 TI - 50-foot walking time: a critical assessment of an outcome measure in clinical therapeutic trials of antirheumatic drugs. AB - Fifty-foot walking time was used in 51 of 187 clinical therapeutic trials of antirheumatic drugs and in only 21 instances was statistical significance reached. Measurement of the 50-foot walking time showed no better performance in long-term trials of SAARDs than in short-term trials of NSAIDs. It is concluded that the 50-foot walking time is a poor outcome measure in rheumatic disease trials, despite a high intra- and interobserver reproducibility. PMID- 3052683 TI - The renal rind sign: a new ultrasound indication of inflammatory disease in the abdomen. AB - The width of the right anterior extrarenal tissue is increased on ultrasound examination in patients with abdominal inflammatory disease. Thickened perirenal fascia associated with acute pancreatitis has previously been reported on computed tomography. A case report has described increased echogenicity of the pararenal space on ultrasound in children with pancreatitis but increased width of the space between the liver and the renal capsule has not hitherto been described in association with inflammatory disease in the abdomen in adults. We have observed it in acute cholecystitis, acute pancreatitis, acute appendicitis, a perforated duodenal ulcer, a leaking anastomosis with a right subphrenic abscess following total gastrectomy and in a patient with septicaemia and liver abscesses. Normal values were obtained in 100 patients without detectable or known disease and were found to be between 1 and 6 mm (mean 2.5 mm) in men and 1 and 5 mm (mean 1.8 mm) in women. The patients with abdominal disease who demonstrated this sign had values ranging from 9-11 mm (mean 10 mm). PMID- 3052681 TI - Effects of anterior hypothalamic lesions and sham-operations on bacterial endotoxin-induced non-specific airway hyperreactivity in vivo and in vitro. AB - 1. In the present study the effects of anterior hypothalamic (AHA) lesions and sham-operations were investigated on the endotoxin-induced airway hyperreactivity in guinea-pigs. Unoperated, sham-operated and AHA-lesioned guinea-pigs were injected intraperitoneally with Escherichia coli endotoxin and the airway reactivity tested four days later in isolated tracheal spirals and in spontaneously breathing anaesthetized animals. Control animals were given sterile saline. 2. Sham-operated control animals demonstrated a diminished responsiveness of the tracheal spirals in vitro and of the lung resistance (delta R1) in vivo to histamine receptor and cholinoceptor-muscarinic agonists as compared to unoperated control animals. 3. AHA-lesioned control animals showed a responsiveness of the respiratory airways in vitro and in vivo between the values of unoperated and sham-operated control animals, suggesting that lesions partially restored the diminished responsiveness. 4. In unoperated and sham operated guinea-pigs, endotoxin administration induced hyperreactivity of the tracheal spirals and delta R1 to histamine receptor and cholinoceptor-muscarinic agonists with respect to the control groups. 5. In AHA-lesioned animals, the endotoxin-induced airway hyperreactivity in vitro and in vivo to histamine receptor and cholinoceptor-muscarinic agonists was absent. PMID- 3052685 TI - Spinal segmentation anomalies: in utero diagnosis by ultrasound. PMID- 3052684 TI - Kock pouch urinary diversion: follow-up by ultrasound. AB - In the post-operative follow-up of 24 patients who received a continent Kock pouch for urinary diversion, several complications were encountered, including hydronephrosis, stone formation and valve dysfunction, resulting in reflux and/or urinary incontinence. After comparing findings on ultrasound with those obtained by Koch pouch cystography, intravenous urography, plain abdominal radiography, Kock pouch endoscopy and operation, we consider ultrasound to be an important technique in the follow-up, especially in non-symptomatic patients. All cases of hydronephrosis and pouch calculi were detected by ultrasound and no false positive findings were encountered in either group. A good correlation is demonstrated between nipple length, as measured by ultrasound, and valve dysfunction, clinically important only for the afferent nipple. PMID- 3052686 TI - Congenital hemihypertrophy with adrenal adenoma and medullary sponge kidney. PMID- 3052687 TI - The expanding infant. PMID- 3052688 TI - Surgical treatment for thyrotoxicosis. PMID- 3052689 TI - Prevention of postoperative deep vein thrombosis. PMID- 3052690 TI - Cyclical mastopathy: a critical review of therapy. AB - Because of a recently completed trial which demonstrated that dietary fat reduction improved the symptoms of cyclical mastopathy, we have carried out a critical review of other studies of treatment for this disorder to determine what, if any, mechanisms of action were shared by effective treatments. Therapies were classified as 'definitely effective' if their effectiveness was demonstrated in at least one randomized placebo-controlled study of women with cyclical mastopathy, 'probably effective' if their effectiveness in relieving breast symptoms was demonstrated in at least one study in women with premenstrual syndrome, and 'probably not effective' if they had not been shown to improve breast symptoms in at least two studies in women with premenstrual syndrome. Bromocriptine, danazol, evening primrose oil, tamoxifen and reduction of dietary fat intake were classified as definitely effective and norethisterone and Bellergal as probably effective. Several therapeutic manoeuvres, including reduction of methylxanthine ingestion and the administration of vitamin E or diuretics, have not been adequately evaluated in women with cyclical breast symptoms. A review of the published reports of the physiological effects of therapies that were definitely or probably effective revealed that these agents acted either to alter serum prolactin levels or lipid metabolism, or both. As evidence exists that prolactin influences lipid metabolism it is postulated that cyclical mastopathy may be a disorder of lipid metabolism. PMID- 3052691 TI - Dosage and duration of tamoxifen treatment for mastalgia: a controlled trial. AB - A controlled trial has been conducted in which 60 women with mastalgia were randomly allocated to receive tamoxifen at a dosage of either 10 mg or 20 mg daily for either 3 or 6 months. All eligible patients had self-rated moderate or severe mastalgia present for at least 6 months, for which no specific therapy had been given for the previous 3 months. End points of the study were pain control, measured by linear analogue scales, relapse rate and side-effects. Pain relief was achieved in 90 per cent of those receiving 10 mg daily and 86 per cent of those given 20 mg daily. The relapse rate was also similar for both dosages, being 48 per cent and 39 per cent respectively and usually occurred within 2-3 months of discontinuing treatment. However, side-effects were reported less frequently among those receiving 10 mg daily (21 per cent versus 64 per cent; chi 2 = 11.1, P less than 0.001). Prolongation of treatment from 3 months to 6 months did not materially improve the response rate (85 per cent versus 90 per cent). Side-effects were similar, as was the relapse rate among the patients receiving the two durations of treatment. The agent proved to be significantly more effective in the relief of cyclical rather than non-cyclical pain (94 per cent versus 56 per cent). Use of tamoxifen for the treatment of mastalgia is still experimental. Nevertheless, for the majority of women with mastalgia, pain relief can be achieved using tamoxifen 10 mg daily, given for a 3-month course. As almost half these patients will develop relapse of breast pain it may be necessary to give longer courses of therapy, although the safety of this more protracted treatment has yet to be determined. PMID- 3052692 TI - Role of Doppler ultrasound flowmetry in the diagnosis of breast lumps. AB - One hundred and five patients with discrete breast lumps were examined with a 10 MHz Doppler ultrasonic flowmeter. Doppler flow signals were analysed on an Angioscan spectrum analyser. Recordings from the opposite normal breast were taken as controls and signals from the two sides compared. In 23 patients signals from the normal breast could not be recorded and therefore results of the remaining 82 patients are reported. These included 39 patients with carcinoma, 20 with fibroadenoma, 12 with cyclical nodularity and 11 with cysts. Malignant lumps exhibited significantly higher peak systolic (S) and minimum diastolic frequencies (D) in comparison to the control breast. Fibroadenoma also had a higher S and D than those of the opposite normal breast. Signals over cysts and cyclical nodularity showed no significant difference from the recordings over the control side. Despite significantly higher systolic and diastolic frequencies in the cancer group in comparison to benign lumps and normal breast, considerable overlap in the values was seen between cancer and other groups. Therefore the patterns on 10 MHz Doppler sonography are not sufficiently specific to discriminate benign from malignant breast lumps. PMID- 3052693 TI - Pyogenic liver abscess: an improvement in prognosis. AB - Forty-six patients with pyogenic liver abscess have been treated at Paul Brousse Hospital between 1966 and 1986. The overall mortality was 24 per cent, all 11 deaths occurring in 24 patients seen prior to 1978 when there was often a considerable delay in the diagnosis of liver abscess (mean 90 +/- 71 days). In seven patients the diagnosis was not made until post-mortem examination. The mainstay of treatment was surgical drainage. Since 1978 high resolution imaging techniques for the liver, and in particular ultrasound, have been available. The diagnostic delay has been significantly reduced (mean 28 +/- 20 days, P less than 0.01). Patients are receiving definitive treatment at an earlier stage in the evolution of the disease process, with fewer established complications prior to treatment (P less than 0.05). Percutaneous drainage under ultrasound control is the preferred initial drainage procedure in high-risk patients. There have been no deaths in 22 patients treated for pyogenic liver abscess since 1978 (P less than 0.001). PMID- 3052694 TI - Retroperitoneal extravasation from perforated duodenal ulcer. PMID- 3052695 TI - Ultrasound scanning by the urologist in outpatient clinics. PMID- 3052696 TI - Wilson's disease and epilepsy. AB - The relationship between Wilson's disease and epilepsy is explored, both in the literature and in a series of 200 cases of Wilson's disease. Details of 44 literature and 14 personal cases of both disorders are presented. The prevalence on December 1, 1986 of epilepsy in the Cambridge series was 6.2%, ten times higher than that of epilepsy in the general population. Seizures in Wilson's disease occur at any stage of the disease, but often begin shortly after the start of treatment. Prognosis of seizures was comparable with the best quoted figures for idiopathic epilepsy: at 7 years 60% of cases had been seizure-free for at least 5 years, and 75% for at least 2 years. Possible mechanisms of seizures are discussed. Penicillamine-induced pyridoxine deficiency is probably not involved in more than a minority of cases. It is more likely that a direct effect of copper deposition is responsible for most of the seizures. PMID- 3052698 TI - Actions of excitatory amino acid acid agonists and antagonists on the primary afferents of the vestibular system of the axolotl (Ambystoma mexicanum). AB - In order to determine the nature of the transmitter in the synapse between hair cells and primary afferent fibers, both resting and evoked spike activity of vestibular system afferents were recorded. Excitatory amino acid agonists and antagonists were applied by micro perfusion. Excitatory amino acid agonists consistently increased the firing rate of these afferents. The rank order in potencies of the agonists tested was: kainate greater than or equal to quisqualate greater than D-aspartate greater than or equal to L-glutamate greater than or equal to L-aspartate greater than N-methyl D-aspartate. Blockade of synaptic transmission with high-Mg2+ and low-Ca2+ solutions did not seem to affect the responses to the excitatory amino acid agonists indicating their postsynaptic action. Excitatory amino acid antagonists inhibit both resting and physiologically evoked activity. The rank order of inhibitory potency was: kynurenate greater than L-glutamate diethyl ester greater than D,L-2-amino-4 phosphono-butyrate greater than D-alpha-amino adipate greater than D,L-2-amino-5 phosphonovalerate. These findings suggest that an amino acid-related compound may be the transmitter at this synapse. The relative potencies of agonists and antagonists tested provide evidence that the transmitter released from the hair cells' basal pole in the axolotl vestibular system interacts with postsynaptic kainic/quisqualic type receptors. PMID- 3052697 TI - Sex differences in the responses of hypothalamic luteinizing hormone-releasing hormone and catecholamine systems to ovarian hormones and naloxone: implications for sexual differentiation of luteinizing hormone secretion in rats. AB - Normal male rats, or female rats exposed neonatally to androgens or estrogens, do not respond in adulthood to ovarian hormone treatments that stimulate preovulatory-like surges of luteinizing hormone (LH) or mating behavior in normal females. As an attempt to understand the neurochemical basis for this insensitivity, the present studies tested whether sex differences also exist with respect to several important neural events that are antecedent to and essential for the appearance of an LH surge induced by ovarian hormone treatment. Administration of estradiol via Silastic capsules to adult, gonadectomized rats resulted in a suppression of LH release that was equivalent in males and females, but only the estrogen-primed females responded to injections of progesterone with an LH surge. Similarly, in estrogen-primed females but not males, progesterone induced a presurge sequential accumulation and decline of LH-releasing hormone (LH-RH) concentrations in the median eminence and increased the turnover rates of norepinephrine (NE) and epinephrine (E) in the medial basal hypothalamus during the time of LH-RH accumulation. Ovarian hormones may activate NE and E release in females by removing a tonic inhibition over catecholamine release exerted by endogenous opioids. In order to test whether direct antagonism of opiate mechanisms would produce equivalent neuroendocrine or neurochemical responses in males and females, additional studies tested the effects of the opiate receptor blocker naloxone on LH release and on activity of catecholamines in the medial basal hypothalamus. In contrast to females, estrogen-primed male rats did not display either an increase in serum LH or an enhancement of the alpha methyltyrosine-induced decline of NE or E after treatment with naloxone.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3052699 TI - [The effect of long-term smoking on the thyroid gland in women]. PMID- 3052700 TI - [Ultrasound-guided fine needle biopsy of the abdominal organs and structures]. PMID- 3052701 TI - [Contribution of the staff of the Pediatric Clinic of Comenius University Medical School to child welfare 1919-1930]. PMID- 3052702 TI - [Combination of computer tomography and stereotaxic technic in the diagnosis and therapy of brain tumors]. PMID- 3052703 TI - The role of high-dose therapy and autologous bone marrow reinfusion in the treatment of malignant lymphomas. AB - In a significant fraction of patients with NHL or HD, disease develops that is resistant to conventional chemotherapy. Experience using high-dose chemotherapy, with or without TBI, and ABMR is expanding. In HD, remissions can be achieved in approximately half of the patients with relapsed advanced disease. This may also be true in patients with NHL who do not respond to conventional regimens. High dose chemoradiotherapy regimens are toxic and require extensive supportive care. Relapse frequently occurs in areas of previous disease, suggesting failure of the conditioning regimen rather that an infusion of occult tumor cells in the autologous bone marrow had occurred. Thus, the role of marrow purging in this therapy needs to be further evaluated and compared with findings involving nonpurged marrow reinfusion. It is also important to evaluate the effects of more vigorous attempts at cytoreduction of bulky disease prior to high-dose therapy and ABMR. We recommend that high-dose therapy and ABMR in an investigational setting be used (1) in patients with HD who experience relapse after MOPP/ABVD or equivalent regimens and (2) in patients with intermediate or high-grade NHL whose disease recurs or is resistant to conventional regimens. Potential areas for development include the use of this modality as intensification therapy following conventional therapy in patients with intermediate or high-grade NHL with poor prognostic features. Toxicity can be decreased and efficacy increased only if therapy is administered to patients who have not been heavily pretreated and who have lower tumor burden and a good performance status. The role of high-dose chemotherapy of ABMR in the nodular lymphomas is not known at this point. Finally, high-dose ABMR therapy has a definite role in salvaging patients with malignant lymphomas. Many issues need to be resolved, including (i) the optimal timing of this approach, (ii) the optimal conditioning regimen, and (iii) the need for purging bone marrow prior to reinfusion. The past 10 years have led to significant gains. During the next 10 years, it may be possible to refine this therapy and find solutions to the above issues. PMID- 3052704 TI - Membranes and cancer chemotherapy. PMID- 3052705 TI - Autologous bone marrow transplantation for patients with advanced lymphomas: improvement of a promising investigational technique. PMID- 3052706 TI - Immune dysfunction in the critically ill infant and child. AB - Factors contributing to the high prevalence of immunodeficiency in the PICU population include conditions that lead to frequent requirement of intensive care, suppression of immunity secondary to an acute insult, and iatrogenic measures. The immunodeficiency observed in the critically ill correlates well with their susceptibility to infection and explains the high prevalence of nosocomial sepsis in the PICU--a major cause of morbidity and mortality in critically ill children. Dysactivation of the immune system during an acute insult, with the subsequent release of humoral mediators from activated immune cells, leads to tissue injury and may be involved in the pathogenesis of ARDS, DIC, capillary leak syndrome, and to the development of multiple organ system failure. Suggested approaches to correct the immunodeficiency in the critically ill include reconstitutional immunotherapy, mediator-inhibiting drugs, and mediator removal by plasma exchange. Intensivists should be aware of the phenomenon of immunodeficiency in the critically ill, be accordingly aggressive in diagnosing and treating infections, and avoid, as much as possible, measures that further suppress immunity. PMID- 3052707 TI - Selected concepts and controversies in pediatric cardiopulmonary resuscitation. AB - Although more than 80 years of research in cardiac resuscitation produced many important findings and greatly enhanced our understanding of the arrest state, outcome following pediatric cardiac arrest remains poor. Resuscitation guidelines have recently been published, but they may not reflect optimal therapy. Closed chest compression-induced cardiac output may be higher in pediatric patients, particularly infants, than that previously reported in adults. To achieve higher cardiac outputs, direct cardiac compression is important; the recommended compression location has therefore been changed based on recent data. The optimal rate of compression, however, is uncertain, so further research is needed. Alternative vascular access sites, such as the endotracheal and intraosseous route for drug administration may permit more rapid drug delivery, but data suggest that a larger epinephrine dose than currently recommended should be used. It may also be helpful to dilute the drug in normal saline before endotracheal administration. Although experimental data suggest that a pure alpha-adrenergic agonist may be beneficial in a cardiac arrest, recent data show that epinephrine remains the drug of choice. Finally, the role of sodium bicarbonate in both the arrest and postarrest setting has become controversial. Recent data suggest that bicarbonate may be detrimental and that therapy of acidosis is best directed at improving perfusion, oxygenation, and ventilation. Alternative forms of therapy for acidosis, such as THAM and dichloroacetate may prove beneficial in the postarrest setting. PMID- 3052709 TI - Issues in airway management--1988. AB - Knowledge of the airway is expanding. New conditions, modern management strategies, and a more complete understanding of the interaction between the airway and fluid flux in the lung are presented. PMID- 3052710 TI - Therapeutic strategies for acute hypoxemic respiratory failure. AB - Acute hypoxemic respiratory failure is a pulmonary capillary leak state that occurs in many different clinical settings. The resultant edema causes refractory hypoxemia due to intrapulmonary shunting and loss of lung compliance. Mechanical ventilation with PEEP and high concentrations of supplemental oxygen are frequently necessary for patient survival, but these therapies may themselves contribute to further lung damage. Studies have shown that manipulations of the pulmonary circulation aimed at reducing the forces promoting edemagenesis have salutary effects on gas exchange and lung mechanics and can be performed safely. The use of these combined modalities for pediatric patient care has been discussed. Finally, other approaches to treatment currently being investigated in animal models of AHRF have been presented. As these and other new treatment methods are explored, hopefully it will be possible to decrease the unacceptably high mortality rate still encountered in AHRF. PMID- 3052708 TI - Bronchopulmonary dysplasia in the pediatric intensive care unit. AB - Bronchopulmonary dysplasia (BPD) is an important cause of chronic respiratory disease in infants and children. Infants with BPD are frequently readmitted to the hospital during the first 2 years of life usually because of infectious exacerbations of their chronic lung disease. This article is a review of the multisystem pathology of BPD and therapeutic approaches to the management of these infants in the PICU. PMID- 3052711 TI - Poisoning. AB - Knowledge of the toxicologic nature of ingested substances provides a proper framework for general and specific therapies best suited to meet the needs of the patient. Monitoring and direct observation provided in the PICU can aid proper therapy for many intoxicants. Good supportive care coupled with specific pharmacotherapy will provide the best chance for a successful outcome. PMID- 3052712 TI - Concern about cyclamate as toothpaste sweetener. PMID- 3052713 TI - Chemotherapeutic products for the control of supragingival dental plaque and gingivitis. PMID- 3052714 TI - Pit and fissure sealants: one more time. PMID- 3052715 TI - The case for fluoridation: a comparison between Toronto and Montreal. PMID- 3052716 TI - Birthplace of Painless Parker revisited. PMID- 3052717 TI - A review of differential diagnosis of dental fluorosis and non-fluoride enamel defects. PMID- 3052718 TI - Nursing caries in the Inuit children of the Keewatin. PMID- 3052719 TI - Comparative effects of FMR programs in fluoridated and unfluoridated communities. PMID- 3052720 TI - "Prosthetics, progress and prudence". PMID- 3052721 TI - New CDA President responds to questions about CDA priorities. PMID- 3052722 TI - Commentary on article 'Porcelain laminates'. PMID- 3052723 TI - Canadian hospital dental department is engaged in HIV and AIDS research. PMID- 3052724 TI - Computers in dentistry. Part one. CAD/CAM: the computer moves chairside. PMID- 3052725 TI - Computers in dentistry. Part two. Francois Duret--a man with vision. PMID- 3052726 TI - Computers in dentistry. Part three. The computer gives new vision--literally. PMID- 3052728 TI - Clinical dental research in Canada. A clinician's perspective. PMID- 3052727 TI - Computers in dentistry. Part four. Bytes 'n bites. The computer moves from the front desk to the operatory. PMID- 3052729 TI - A single dose study of a new analgesic. Ciramadol. PMID- 3052730 TI - Extrusion of endodontic filling material into the inferior alveolar canal. PMID- 3052731 TI - Long-term comparison of two surgical flap designs for third molar surgery on the health of the periodontal tissue of the second molar tooth. PMID- 3052732 TI - [Sonographic appearance of adenomyomatosis of the gallbladder--a case report]. PMID- 3052733 TI - Tuberous sclerosis with unusual rib deformity and abdominal aortic aneurysm--a case report. PMID- 3052734 TI - Symptomatic pericardial effusion in cancer patients. PMID- 3052735 TI - Reconstructive procedures in heart valve disease. AB - The percentage of patients who undergo valve repair has increased considerably during the past decade. Valve repair is particularly important in patients with atrioventricular valve disease since it provides better results than replacement. Aortic valve repair is presently performed only in selected patients with aortic stenosis and insufficiency. There is a renewed interest in aortic valve debridement in patients with calcific aortic stenosis. Mitral valve repair can be performed in most patients with mitral insufficiency and in many with mitral stenosis. The surgical techniques for mitral valve repair are well established and are reproducible. The tricuspid valve is almost always reparable in patients with tricuspid disease associated with mitral valve disease. PMID- 3052736 TI - Valve surgery in children. AB - Valve surgery constitutes a small percentage of overall pediatric cardiac practice. Most of the lesions seen are congenital anomalies, often in association with other lesions. Valvotomy and valvuloplasty can usually provide good long term palliation, although reoperation is probably inevitable. The late results of valve replacement in children have been compromised by the rapid calcification of biologic valves. There is some evidence to suggest that this may not occur with homograft valves. The importance of implanting as large a valve as possible is emphasized and follow-up must include an assessment of whether the child is outgrowing the previously implanted prosthesis. PMID- 3052737 TI - Coronary prevention and regression studies updated. AB - There have been a multitude of clinical, animal and epidemiology studies which prove that high serum cholesterol and low density lipoprotein (LDL) cholesterol concentrations are specific causes of coronary artery disease (CAD). Although the variations in experimental design make comparisons difficult, the aggregate results of many human prevention trials since 1960 lead to the definite conclusion that a 10% lowering of serum cholesterol reduces the risk of CAD by one-sixth. Recently, other factors for CAD risk have been identified that will be useful in guiding treatment, namely serum high density lipoprotein (HDL) cholesterol, apolipoproteins B and AI, HDL triglycerides and phosphatidylcholine to free cholesterol ratio. Studies have shown that aggressive drug and diet therapy slows progression and causes regression of atheromas. Primary prevention of CAD is obviously preferable to secondary prevention. Also, the evidence to date indicates that prevention of CAD through lifestyle changes should begin in childhood. PMID- 3052738 TI - European Consensus on Primary Prevention of Coronary Heart Disease. AB - The European Consensus on Primary Prevention of Coronary Heart Disease has recommended that providing care for individuals at particular risk for coronary artery disease (CAD) requires case finding through medical examinations in primary care, hospital and employment health examination settings. Decisions concerning management of elevated lipid levels should be based on overall cardiovascular risk. The goal of reducing cholesterol levels through risk reduction can ultimately be accomplished only with the implementation of health education efforts directed toward all age groups and actions by government and supranational agencies, including adequate food labelling to identify fat content, selective taxation to encourage healthful habits and wider availability of exercise facilities. Only measures directed at the overall population can eventually reach the large proportion of individuals at mildly to moderately increased risk for CAD. The European Policy Statement on the Prevention of Coronary Heart Disease recognizes that the question of lipid elevation as a risk factor for CAD involves assessment, not only of cholesterol level alone, but also of triglycerides and the HDL cholesterol lipid fraction. Five specific categories of dyslipidemia have been identified, with individualized screening and treatment strategies advised for each. It is the consensus of the study group panel members that these procedures are both practical and feasible. They begin the necessary long term process to reduce the unacceptably high levels of morbidity and mortality due to CAD throughout the European community. PMID- 3052739 TI - Identification of the patient at risk in the physician's office and drug management of dyslipoproteinemia. AB - The wealth of convincing evidence favouring the major role of lipids in atherosclerosis and the benefit of lipid lowering therapy for its prevention now sets the stage for more practical questions: "Whom to treat? When to treat? How to treat?". Although each patient must be considered individually, there are general rules and specific guidelines that should be underlined. An isolated finding of abnormal lipid levels should by no means be a systematic starting point for initiating treatment. In a step by step approach, guidelines for individual risk assessment, systematic search for specific clinical clues and pertinence of complementary investigative measures are given. In addition to ensuring a correct diagnosis, these recommendations will orient the physician's decision with respect to the necessity of initiating treatment and, if warranted, the choice of an optimal treatment algorithm. Therapeutic modalities are discussed both in general and specific terms, focusing mostly on pharmacologic agents. Clinical indications, mechanisms of action, adverse effects and expected efficacy are seen for each drug or family of drugs, including those with a promising future. In summary, 10 easy rules of thumb are provided to ensure adequate diagnosis and management of hyperlipidemia. PMID- 3052740 TI - Immunogenicity of monocomponent human and porcine insulin in non-insulin dependent diabetes mellitus. PMID- 3052741 TI - Diprosopus--a case report. PMID- 3052742 TI - [Biliary enteric fistula]. PMID- 3052743 TI - [The clinical application of autologous human cultured epithelia]. PMID- 3052744 TI - Usefulness of computerized visual acuity testing in a pediatric ophthalmology clinic. AB - We investigated the usefulness of a computerized illiterate-E-style acuity test in 111 children ranging in age from 3 to 13 (mean 6.56) years consecutively referred to a large pediatric ophthalmology clinic. Visual acuity was measured in both eyes of each subject by means of conventional testing with an optical projection device and computerized testing with a program run on a microcomputer. Of the 222 eyes 155 (70%) showed no difference in visual acuity between the two procedures. A difference of one acuity line was found for 63 eyes (28%), and a difference of two lines was found for 4 eyes (2%). The conventional test yielded a higher acuity score in 75% of the cases in which a one-line difference was found and 50% of those in which a two-line difference was found. Our results indicate that computerized testing is a valid and effective method of assessing visual acuity in children. PMID- 3052745 TI - Expression of transforming growth factor alpha during development. AB - Transforming growth factors (TGFs) are polypeptides that are produced by transformed and tumour cells, and that can confer phenotypic properties associated with transformation on normal cells in culture. One of these growth regulating molecules, transforming growth factor alpha (TGF-alpha), is a 50 amino acid polypeptide that is related to epidermal growth factor (EGF) and binds to the EGF receptor. Previous studies have shown that TGF-alpha is expressed during rodent embryogenesis between 7 and 14 days gestation. To investigate the cellular sites of TGF-alpha mRNA expression during development, we have performed Northern analyses and in situ hybridization histochemistry on the conceptus and maternal tissues at various gestational ages. Contrary to previous reports, both Northern analyses and in situ hybridization histochemistry indicate that TGF-alpha mRNA is predominantly expressed in the maternal decidua and not in the embryo. Decidual expression is induced following implantation, peaks at day 8, and declines through day 15 when the decidua is being resorbed. In situ hybridization revealed that expression of TGF-alpha mRNA is highest in the region of decidua adjacent to the embryo and is low or nondetectable in the uterus, placenta, and embryo. In addition, we could not detect TGF-alpha mRNA expression in other maternal tissues, indicating that the induction of TGF-alpha transcripts in the decidua is tissue specific, and not a pleiotropic response to changes in hormonal milieu that occur during pregnancy. The developmentally regulated expression of TGF alpha mRNA in the decidua, together with the presence of EGF receptors in this tissue, suggests that this peptide may stimulate mitosis and angiogenesis locally by an autocrine mechanism.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3052746 TI - Physiological factors in fetal lung growth. AB - Adequate pulmonary function at birth depends upon a mature surfactant system and lungs of normal size. Surfactant is controlled primarily by hormonal factors, especially from the hypophysis, adrenal, and thyroid; but these have little influence on fetal lung growth. In contrast, current data indicate that lung growth is determined by the following physical factors that permit the lungs to express their inherent growth potential. (a) Adequate intrathoracic space: lesions that decrease intrathoracic space impede lung growth, apparently by physical compression. (b) Adequate amount of amniotic fluid: oligohydramnios retards lung growth, possibly by lung compression or by affecting fetal breathing movements or the volume of fluid within the potential airways and airspaces. (c) Fetal breathing movements of normal incidence and amplitude: fetal breathing movements stimulate lung growth, possibly by stretching the pulmonary tissue, and do not affect mean pulmonary blood flow but do induce small changes in phasic flow; these changes are probably too slight to influence lung growth. (d) Normal balance of volumes and pressures within the potential airways and airspaces: in the fetus, tracheal pressure greater than amniotic pressure greater than pleural pressure. This differential produces a distending pressure which may promote lung growth. Disturbing the normal pressure relationships alters the volume of fluid in the lungs and distorts lung growth, which is stimulated by distending the lungs and is impeded by decreasing lung fluid volume. The mechanisms by which these factors affect lung growth remain to be defined. Fetal lung growth also depends on at least a small amount of blood flow through the pulmonary arteries.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3052748 TI - Issue dedicated to Professor R.G.E. Murray in recognition of his contributions to microbiology. PMID- 3052747 TI - The control of cardiovascular shunts in the fetal and perinatal period. AB - The fetal circulation has two major vascular shunts, the ductus arteriosus and the ductus venosus. The ductus arteriosus connects the pulmonary artery with the descending portion of the aortic arch, hence shunting most of the right ventricular output away from the unexpanded lungs. The ductus venosus connects instead the portal sinus with the inferior vena cava and allows well-oxygenated umbilical vein blood to bypass the liver and reach the central circulation rapidly. Both blood vessels cease their function after birth and undergo permanent closure. It is now well established that prenatal patency of the ductus arteriosus is an active state sustained by a prostaglandin. A similar mechanism has been recently recognized in the fetal ductus venosus. Evidence is presented indicating that prostaglandin E2 and prostaglandin I2 are natural relaxants, respectively, for the ductus arteriosus and the ductus venosus. In addition, both vascular shunts share the dependence on an endogenous cytochrome P-450 mechanism to develop their contractile tone. This mechanism may be important in the normal process of shunt closure at birth. While broadening the knowledge of fetal cardiovascular homeostasis, advances in this field have important implications for the prevention and management of certain pathological conditions affecting the newborn. PMID- 3052749 TI - The bacterial surface: general considerations towards design and function. AB - The shape of bacteria is determined by the wall which surrounds the protoplast. This enveloping structure resides in diverse chemical and organizational forms and is of fundamental importance for several aspects of cellular life and growth. This article discusses these general aspects and serves as an introduction to the journal issue and to the more specific papers which follow in the section. PMID- 3052750 TI - Growth and form in microorganisms: morphogenesis of Escherichia coli. PMID- 3052751 TI - Speculations on the growth strategy of prosthecate bacteria. AB - Appendaged bacteria with stalks that are extensions of the cell wall have had to solve the problems of growing the stalk as a tube of constant diameter and of partitioning their chromosomes into the asymmetric daughter cells. Although no experimental proof is given, it is suggested that both processes depend on the attachment of the chromosome origin and terminus to the wall at special terminal sites that contain the basal body (motor assembly) for flagellar motion. PMID- 3052752 TI - Bacterial extracellular polysaccharides. AB - The synthesis of extracellular polysaccharides has been recognized in certain bacterial cultures since the 1880s. It is now apparent that a wide range of bacteria produce these polymers and an equally wide range of chemical structures are possible. Their surface location, together with the range of available monosaccharide combinations, noncarbohydrate substituents, and linkage types, make extracellular polysaccharides excellent agents of diversity. As a result, much effort has been directed towards elucidating their structure in pathogenic bacteria and in enteric organisms in particular. Commercial applications of microbial polysaccharides have now broadened the scope of structural information. Most recently, technological advances in molecular biology have created the possibility of manipulating desired polymer characteristics, and with these advances, our knowledge of the mechanisms of synthesis and regulation of cell surface polysaccharides has improved. Ultimately more information regarding the function of extracellular polysaccharides in natural environments may result. PMID- 3052753 TI - The domains of slow bacterial growth. AB - Commonly used culture systems, e.g., batch culture and the chemostat, work poorly for defining the behavior of slowly growing bacteria, i.e., cultures with mass doubling times, tD, longer than 10-12 h. This has slowed the process of understanding how bacteria behave at the longer tD's that embrace most of their existence. Culture systems are identified that give useful access to these longer doubling times. When these slow-growth systems are used with nutrient-limited populations, it is found that cellular concentrations of guanosine 5'-diphosphate 3'-diphosphate, the main effector of the stringent response, commence rising above basal levels at tD's longer than 12 h until, at a tD of 60-70 h, the level is reached that causes the interdiction of protein and ribosome synthesis characteristic of the response. The abrupt onset of the stringent response in eubacteria at a particular tD, corresponding to a specific rate of nutrient uptake, makes oft-used growth equations that do not account for this onset, e.g., those of Monod or Pirt, poor descriptors of slow growth. A serious imbalance between lateral and transverse wall formation rates which, unless corrected, would lead to abnormal cell division can be inferred to develop at tD's longer than 10-12 h. The necessary correction may be supplied by the effects of increased levels of guanosine 5'-diphosphate 3'-diphosphate on transverse wall synthesis. PMID- 3052754 TI - Bacterial flagellar structure and function. PMID- 3052755 TI - The flagellar filament protein. PMID- 3052756 TI - The bacterial chemosensory system. PMID- 3052757 TI - Microbial invasion: a covert activity? AB - In contrast to nonpathogenic microorganisms that exist happily in biofilms on various organic and inorganic surfaces, many pathogenic microbes have the additional ability to invade host tissues by inducing their own endocytosis and transport across normally protective barriers. This phenomenon, designated "parasite-directed endocytosis," has been observed with a variety of surfaces (intestinal, genital, nasopharyngeal, and tracheal epithelium) as well as in endothelial cells. The mechanisms involved in invasion may involve a single factor as described for some species of Yersinia, or may require multiple factors as observed in Shigellae. For the majority of pathogens, the molecular mechanisms of invasion are not well understood (e.g., Neisseria gonorrhoeae). Because parasite-directed endocytosis is reminiscent of receptor-mediated endocytosis, it is quite possible that some pathogens engage in biologic mimicry by producing a molecule that resembles a natural host ligand, for which there is a host cell receptor. Such a masquerade may allow some microbes to enter the host's inner sanctum covertly in a manner analogous to the Trojan horse, rather than overtly by destroying the mucosa and entering host tissues directly. Whereas this hypothesis is speculative at present, bacteria that produce molecules resembling insulin, calmodulin, and chorionic gonadotropin have been described. PMID- 3052758 TI - Structure and plasticity at various organization levels in the bacterial cell. PMID- 3052759 TI - Aspects of plasmid F maintenance in Escherichia coli. AB - A major class of replicons in procaryotes is typified by low copy number, nonrandom intracellular distribution, and stable inheritance. Included in this class are chromosomes of gram-positive and gram-negative bacteria as well as a number of plasmids from these organisms. Replicons in this major class have remarkable structural and functional similarities in the genes that effect and control replication. In the present work a review of plasmid F is presented as a paradigm for many aspects of this group's maintenance features. PMID- 3052760 TI - Bacterial evolution. AB - The deliberate application of the methodology of contemporary molecular genetics to problems in bacterial systematics has led to a broad new understanding of the evolutionary history of both prokaryotes and eukaryotes. In this review, I discuss some of the major conclusions of this endeavour and try to predict future directions. PMID- 3052761 TI - Phylogenetic relationships vs. phenotypic diversity: how to achieve a phylogenetic classification system of the eubacteria. AB - To establish a hierarchic classification system, ranks cannot be defined by the exclusive and inflexible application of phylogenetic parameters. Because both stability and practicality are prerequisites for a successful system, decisions about the delineation of genera must be made by combining phylogenetic coherency with unifying phenotypic properties of taxonomic value consistent with the needs of a hierarchic system. The phylogenetic depth (age) of a genus has no influence on the decision as long as the members of the genus can be reliably identified as such. The description of those higher taxa that are not easily definable today because of the lack of common phenotypic properties must be postponed until new insights are available. In the end this approach will be both phylogenetic and practical, thus avoiding the use of two classification systems. PMID- 3052762 TI - Diabetes and the surgeon. PMID- 3052763 TI - Current perspectives in the management of soft-tissue sarcoma. The role of chemotherapy in multimodality therapy. AB - The role of chemotherapy in adult soft-tissue sarcomas is controversial. In this review, the author examines the effectiveness of single-agent and combination chemotherapy to manage advanced disease and the role of adjuvant combination chemotherapy to control primary tumours and prevent the spread of the disease. Doxorubicin (Adriamycin) is still considered the most effective single agent for advanced soft-tissue sarcoma. Ifosfamide has given similar results and may have greater potential in combination therapy than cyclophosphamide. A current study using trimetrexate shows early positive results. Doxorubicin, cyclophosphamide, vincristine and dacarbazine is currently the most efficacious combination. The efficacy of adjuvant chemotherapy remains to be established. The majority of studies indicate some benefit with chemotherapy, particularly in the relapse-free survival rate, but no consistent improvement in overall survival has been noted. PMID- 3052764 TI - Limb-sparing management in extremity soft-tissue sarcomas in the adult: the radiation oncologist's viewpoint. AB - Current management of extremity soft-tissue sarcomas achieves local control in a high proportion of patients. This paper describes the local methods to conserve limb function that have largely replaced traditional approaches of ablative surgery. Frequently, these methods employ conservative surgery with preoperative or postoperative radiotherapy. An understanding of the biology and behaviour of these rare tumours is necessary to optimize the goals of limb conservation without compromising local control. Appropriate surgical and radiotherapy techniques are facilitated by planned pretreatment staging methods and communication between the surgical and radiation oncology teams. Although a high rate of local control can be expected, distant metastases continue to be a problem. Future strategies should be directed towards scheduling of optimal local treatments with the investigation of adjuvant systemic methods to reduce the rate of distant metastases. PMID- 3052765 TI - Unusual gastric foreign body: a case report. AB - Ingestion of foreign bodies, either intentionally or accidentally, is quite common. The authors report an unusual case in which a 32-year-old man deliberately assembled a large metallic aggregate in his stomach by swallowing magnets and coins in order to relieve epigastric discomfort. The collection was retrieved by laparotomy and gastrotomy; a gastric ulcer was also found and it was oversewn. Management of this patient is discussed and the principles of treatment for ingested foreign bodies are reviewed. PMID- 3052766 TI - Acute Pancoast's syndrome caused by fungal infection. AB - Nonmalignant causes of Pancoast's syndrome are extremely rare. The authors report the case of a 32-year-old man, receiving treatment for acute lymphoblastic leukemia, who had a clinical picture resembling that of Pancoast's syndrome. Invasive mucormycosis was diagnosed as the cause of the syndrome at emergency thoracotomy undertaken to control massive hemoptysis. In spite of adequate treatment, the patient died 5 weeks postoperatively of overwhelming sepsis. A review of the literature disclosed only two other similar cases. The authors conclude that the development of Pancoast's syndrome in the immunosuppressed patient should raise suspicion of an invasive fungal infection. A precise early diagnosis may allow successful, specific antifungal therapy to be instituted. PMID- 3052768 TI - Do not forget MDs' wartime sacrifices, CMA president urges. PMID- 3052767 TI - Doctor or Mister (the correct appellation of British surgeons) AB - The form of address for British surgeons--"Mister" instead of "Doctor"--has mystified other members of the medical profession for years. The author attempts to show that the designation "Mister" is neither an affectation nor a denigration but a natural consequence of the history of British barbery, barber-surgery and ultimately surgery, resulting from the advice and tutelage of King Henry VIII and Parliament. PMID- 3052769 TI - Trimethoprim-sulfamethoxazole v. amoxicillin in the treatment of acute otitis media. AB - Although amoxicillin has long been the preferred drug for treatment of acute otitis media, resistant strains of two relatively common causal organisms have emerged, prompting a search for other antibiotics. We performed a randomized double-blind trial comparing amoxicillin and trimethoprim-sulfamethoxazole in 221 children in whom acute otitis media was diagnosed in an outpatient setting. Diagnosis was on the basis of symptoms, otoscopic examination and acoustic reflectometry. No culture specimens were taken. A research nurse, using the same methods, evaluated patients in a follow-up home visit at around 14 days and measured compliance by examination of the medicine bottle. Equal proportions of children in the two groups were cured or improved (88% and 87%). Therapeutic efficacy was related to compliance in both groups, and there were few side effects in either group. This study had statistical power of 80% to detect a difference of 15%. We conclude that trimethoprim-sulfamethoxazole can be considered a first-line antibiotic in the treatment of acute otitis media. PMID- 3052770 TI - When the emperor is ill, the Japanese take notice. PMID- 3052772 TI - Growth hormone treatment and leukemia. PMID- 3052771 TI - Cranial dystonia, blepharospasm and hemifacial spasm: clinical features and treatment, including the use of botulinum toxin. AB - Blepharospasm, the most frequent feature of cranial dystonia, and hemifacial spasm are two involuntary movement disorders that affect facial muscles. The cause of blepharospasm and other forms of cranial dystonia is not known. Hemifacial spasm is usually due to compression of the seventh cranial nerve at its exit from the brain stem. Cranial dystonia may result in severe disability. Hemifacial spasm tends to be much less disabling but may cause considerable distress and embarrassment. Patients affected with these disorders are often mistakenly considered to have psychiatric problems. Although the two disorders are quite distinct pathophysiologically, therapy with botulinum toxin has proven very effective in both. We review the clinical features, proposed pathophysiologic features, differential diagnosis and treatment, including the use of botulinum toxin, of cranial dystonia and hemifacial spasm. PMID- 3052773 TI - Bronchial challenges. PMID- 3052774 TI - Diagnostic tests for food allergy. PMID- 3052776 TI - Concepts and clinical application of fiberoptic examination of the upper airway. PMID- 3052775 TI - Nasal provocation. PMID- 3052777 TI - Otitis media. AB - Otitis media with effusion is a common disorder in children and occurs more frequently in adults than has been generally appreciated. Laboratory tools have permitted a more specific characterization of this disorder by objective means. These laboratory tests include specific ear tests (pneumatic otoscopy, pure tone, and impedance audiometry), nasal tests (inflammatory cytology), functional tests for obstruction (airway resistance measurements), and anatomic tests of obstruction and inflammation (radiographic of nose and paranasal sinuses). Immunologic tests define the presence of hypersensitivity (skin testing, RAST) and immunodeficiency in this disorder. PMID- 3052778 TI - Laboratory diagnosis of the allergic eye. PMID- 3052779 TI - Treatment of hepatic epithelioid hemangioendothelioma with liver transplantation. AB - Ten patients received liver transplants for unresectable epithelioid hemangioendothelioma (EHE). At the time of transplantation, four patients had microscopic metastases to the hilar lymph nodes, and one of the four also had metastases to a rib. The fifth patient had metastases to the lung, pleura, and diaphragm. The remaining five patients were believed to be free of metastatic disease. Two of these five patients died of metastatic disease at 3 and 16 months, respectively, after transplantation. Interestingly, all five patients with metastatic involvement are currently alive 40.6 +/- 22 months (mean +/- standard error of mean [SEM]) after transplantation, although one of these patients currently has metastatic disease to the lungs and mediastinum. Thus, the projected 5-year actuarial survival rate is 76%, with two patients at risk after the third year. In conclusion, liver transplantation is a reasonable procedure for bulky, otherwise unresectable, EHE even in the presence of metastatic disease. PMID- 3052780 TI - Expression of c-myc oncogene product and ras family oncogene products in various human malignant lymphomas defined by immunohistochemical techniques. AB - The authors studied the expression of c-myc and ras family oncogene products in 43 cases of malignant lymphoma (ML) using the immunoperoxidase method. Unfixed frozen sections of lymph nodes from four patients with Hodgkin's disease and 39 with non-Hodgkin's lymphoma, together with normal lymph nodes, were studied by the avidin-biotin-peroxidase complex (ABC) technique. Two monoclonal antibodies, MYC-2 raised against recombinant human c-myc protein (reacting specifically with the c-myc products P62 and P67) and RASK-4 (raised against recombinant P21 and reacting specifically with ras-family product P21) were used. The c-myc product was detected in nuclei of ML cells and some normal, mainly germinal center, lymphocytes. When the staining intensity shown by normal germinal-center lymphocytes was graded as positive (+) or weakly positive (+/-), a very intensely positive reaction ( to ++) was observed in 37 cases (86%) of ML, a positive reaction (+) in four cases (9.3%), and a weakly positive reaction (+/-) in two cases (4.7%). The ras family oncogene product reaction was intensely positive (++) in two cases (4.7%), positive (+) in 16 cases (37.2%), weakly positive (+/-) in 13 cases (30.2%), and negative in 12 cases (27.9%). Western blot analysis confirmed an elevated level of c-myc products in two cases, which showed intense MYC-2 staining, and of ras family products in one case, which demonstrated intense RASK-4 staining. The enhanced expression of these gene products may play an important role in lymphomagenesis of such cases. PMID- 3052782 TI - Epidermal growth factor receptor in meningiomas is expressed predominantly on endothelial cells. AB - The immunohistochemistry of the epidermal growth factor receptor (EGFR) was studied with monoclonal antibodies in 12 meningiomas of various histologic subtypes, nine benign and three malignant. Strong immunoreactivity of EGFR epitopes was found in the endothelia of the tumor vasculature in six cases. A much weaker reaction was detected within tumor cells in six cases, in one of which it was diffuse and five focal. No correlation was established between the presence of EGFR epitopes and the histologic type or biologic behavior of the meningiomas. The results suggest that the EGFR may participate in tumor angiogenesis, but its role in the growth of neoplastic meningioma cells remains elusive. PMID- 3052781 TI - Miliary pulmonary neuroblastoma. A risk of autologous bone marrow transplantation? AB - A case of miliary pulmonary neuroblastoma is described which occurred 8 months after autologous bone marrow transplantation for Stage IV neuroblastoma. The patient, a 4-year-old boy had pulmonary symptoms at this time. Disseminated metastatic disease was found, but there was no tumor at the primary site. This unusual pattern of relapse suggests that the patient's disseminated disease was related to infusion of malignant cells at the time of transplantation. PMID- 3052783 TI - Adenocarcinoma arising in vulvar breast tissue. AB - A patient with adenocarcinoma arising in vulvar ectopic breast tissue is described. Hormonal receptors of the tumor are analyzed. This is the first case in which an intensive treatment with combined surgery, radiation, hormonal therapy, and cytotoxic chemotherapy was given. Five cases reported previously in the literature are reviewed. PMID- 3052784 TI - Mediastinal osteosarcoma with extension to lungs in a patient treated for Hodgkin's disease. AB - This article reports a patient with Hodgkin's disease in remission after combined modality therapy who developed metastatic pulmonary osteosarcoma, subsequently found to originate in soft tissue of the mediastinum, within a field irradiated 11 years previously. This patient developed a series of radiotherapy-induced complications in addition to osteosarcoma. Only 16 cases of extraskeletal osteosarcoma after radiation treatment have been reported, none originating in the mediastinum. To the authors' knowledge, this is the first reported case of extraskeletal osteosarcoma occurring in a patient previously treated for Hodgkin's disease and with the sarcoma originating within the irradiated field. PMID- 3052786 TI - Metastatic retinoblastoma successfully treated with immunomagnetic purged autologous bone marrow transplantation. AB - There are no documented cases of long-term, disease-free survival in retinoblastoma (RB) metastatic to the bone marrow. The following study details successful outcome in a 3-year-old child with extensive marrow replacement 7 months postenucleation in an otherwise untreated group V RB. Therapy consisted of combinations of vincristine, doxorubicin, and cyclophosphamide for 3 months. After demonstrating cross-reactivity by a panel of six monoclonal neuroblastoma (NB) antibodies with the patient's RB cells, her marrow was purged by using microsphere-linked monoclonal antibodies, and then reinfused as rescue therapy after ablative doses of etoposide and cyclophosphamide. The authors conclude that short-term induction therapy followed by marrow ablative combination chemotherapy and immunomagnetically purged autologous marrow rescue can (1) effect successful outcome in widely metastatic RB, and (2) eliminate the risk of therapy-induced second malignancies. PMID- 3052785 TI - A prospective, randomized double-blind trial comparing metoclopramide alone with metoclopramide plus dexamethasone in preventing emesis induced by high-dose cisplatin. AB - We observed 50 patients receiving high-dose cisplatin-based chemotherapy in a prospective, randomized double-blind trial. One group received metoclopramide (MCP) alone (total dose, 6 mg/kg), whereas the other group was given dexamethasone (DMS) (total dose, 60 mg) in addition to MCP. The patient characteristics of the two groups were comparable, confirming satisfactory randomization. Multivariate regression analysis failed to show any statistical significance in the antiemetic response between the two treatment groups. However, female patients receiving Adriamycin (Adria Laboratories, Columbus, OH) concurrently and obese persons exhibited more vomiting. The overall antiemetic response rate was 66%. Because the side effects were minimal, a higher dose of MCP is expected to improve emetic control without increasing toxicity. The use of a 36-hour assessment period in our study gave more meaningful data. An exponential increase in the dose of MCP is probably required, with respect to weight, to obtain the same antiemetic efficacy. PMID- 3052787 TI - Kinetic analysis of prostatic volume in treating prostatic cancer and its predictability for prognosis. AB - Prostatic volume in patients with prostatic cancer diminished remarkably after castration according to measurements by transrectal sonography. These kinetic changes in prostatic volume were analyzed and a formula was applied to the regression curve. The reduction time (tau) in the formula, which is a constant representing the time interval required for the effective portion of the prostate to be reduced to one tenth, was thought to be a good index to predict the prognosis of the patients. PMID- 3052788 TI - Incontinentia pigmenti, a chromosomal instability syndrome, is associated with childhood malignancy. AB - Incontinentia pigmenti (IP) is a rare hereditary disorder that has recently been classified as a chromosomal instability syndrome. As in Fanconi anemia and ataxia telangiectasia, spontaneous and inducible chromosomal aberrations primarily of the chromatid type are increased in patients with IP. Both Fanconi anemia and ataxia telangiectasia are genetic diseases that predispose to cancer. A case report of an infant with IP and malignancy (rhabdoid tumor of the kidney) is presented, and five previously reported cases of this association are reviewed. The malignancies in all of these cases occurred before age three, whereas malignancy associated with Fanconi anemia and ataxia telangiectasia tends to appear in late childhood or in adulthood. The chromosomal instability seen with IP may increase the risk for malignancy in young children. PMID- 3052789 TI - The association of progressive, atrophying, chronic, granulomatous dermohypodermitis with Hodgkin's disease. AB - The case of a patient with an unusual skin disorder--progressive, atrophying, chronic, granulomatous dermohypodermitis (PACGD)--who developed Hodgkin's disease is reported. A review of the literature revealed only two other cases of PACGD, one of which affected a patient who also was found to have Hodgkin's disease. In an additional report, the diagnosis of Hodgkin's disease was made in a patient who may have had the same dermatologic disorder. The case is reported because the association of these two rare diseases is believed to be more than a chance event. PMID- 3052791 TI - Angiosarcoma associated with foreign body material. A report of three cases. AB - The production of tumors through solid-state mechanisms has been demonstrated in experimental animals, but foreign body tumorigenesis has not been proven definitively in man. The authors report three patients with angiosarcoma that occurred in intimate association with foreign material retained for prolonged periods. Although several etiologic factors have been defined in angiosarcoma, foreign bodies generally are not appreciated to have this potential. Review of the literature disclosed six cases of angiosarcoma and 40 cases of sarcomas of other histologic types associated with foreign material, with latency periods of from 4 months to 63 years. Implanted foreign material thus should be considered capable of inducing virtually any form of sarcoma in humans. PMID- 3052790 TI - Simultaneous gastric cancer in monozygotic twins. AB - Monozygotic twins developed gastric cancers that were found almost simultaneously. A 47-year-old man complained of nausea and vomiting; an upper gastrointestinal series and endoscopy revealed advanced gastric cancer invading the serosa. Palliative subtotal gastrectomy was performed. In his asymptomatic twin a gastric polyp was detected during a screening examination, and this was observed for 2 years. After the former twin had undergone surgery, the latter twin was given a detailed endoscopic examination, and biopsy revealed gastric cancer limited to within the mucosa. Curative subtotal gastrectomy was performed. The noncancerous gastric mucosa of the former twin showed severe intestinal metaplasia, but that in the latter showed only spotty metaplasia. They had lived together for 40 years, but the former was a heavy smoker and drank alcohol, while the latter did not. These differences in taste might have contributed to the observed difference in intestinal metaplasia, which indicates chronic mucosal damage. PMID- 3052793 TI - Approaches to MR angiography. AB - One of the most revolutionary recent imaging advances is the use of magnetic resonance to study and produce morphologic representations of flowing blood vessels known as MR angiography. The ability to produce an image of even moderate spatial resolution of the three dimensional course of blood vessels with MR could have significant advantages over conventional invasive angiography which requires ionizing radiation and contrast material injection. By definition, MR angiography does not require the addition of any intravascular contrast agents and the images are produced entirely by the effect of the radio frequency pulses and magnetic field gradients on the spinning protons. Several researchers are already producing relatively high resolution MR angiograms using a variety of techniques. Essentially all techniques of MR angiography use variations of three steps to produce the image: (1) a projection image, (2) suppression of background static material, and (3) production of a flow sensitive image. This report will survey some of the more commonly used approaches to MR angiography that are currently under investigation. PMID- 3052792 TI - Analysis of early infectious complications after autologous bone marrow transplantation. AB - We reviewed the hospital course of 35 patients who underwent autologous bone marrow transplantation. Fever and profound neutropenia developed in all. Microbiologically confirmed infection developed in 22 patients, and unconfirmed but clinically evident infection developed in six. A bacterial infection developed in 21 patients (most commonly bacteremia without a detectable focus). Mucocutaneous fungal (12 patients) and viral (13 patients) infections were common, whereas invasive fungal (two patients) and viral (one patient) infections were uncommon. New pulmonary infiltrates developed in seven patients. Six deaths occurred during the initial hospitalization for transplantation, only one of which was directly attributable to infection. Stepwise logistic regression analysis retained male gender, total body irradiation, administration of trimethoprim/sulfamethoxazole, and development of mucositis or diarrhea as predictors of decreased survival, whereas higher pretreatment albumin levels and the administration of oral nonabsorbable antifungals were associated with an increased likelihood of survival. A comparison of these infectious complications with those found in allogeneic bone marrow transplant recipients shows similarities and differences with potentially important implications for patient management. PMID- 3052794 TI - Computerized tomographic diagnostic aspects of acquired immunodeficiency syndrome. AB - The involvement of the CNS in patients with AIDS is frequent such that 80 percent of patients have evidence of CNS disease at autopsy. The clinical manifestations are quite varied. These disorders may be divided into two groups: (1) direct effect of the human immunodeficiency virus (HIV) on the nervous system, for example, aseptic meningitis, subacute encephalitis, granulomatous angitis; (2) indirect effects of HIV on the nervous system which are due to immunodysfunction. In the later group, CNS infections (cerebral toxoplasmosis, herpes simplex encephalitis, neurosyphilis, fungal infections, tuberculosis) and unusual CNS neoplasms (Kaposi's sarcoma, primary and secondary brain lymphomas) are the disorders most commonly involving the nervous system. In many of these disorders, the computerized tomographic (CT) findings are helpful in detecting the presence of the abnormality; however, the CT findings are not sufficiently characteristic to permit specific pathological diagnosis without utilization of brain biopsy or CSF findings (including cytological examination and extensive studies for infectious agents). The most common nervous system conditions which complicate AIDS are reviewed emphasizing the conditions in which computerized tomographic findings are most helpful in delineating the pathological condition. PMID- 3052795 TI - Cerebral mucormycosis: a case report. AB - Mucormycosis is an unusual complication in immunosuppressed and diabetic patients. Disseminated mucormycosis is even more unusual. One patient with both pulmonary and cerebral mucormycosis is presented. The clinical course is outlined and computed tomographic (CT) findings are presented. PMID- 3052796 TI - CT diagnosis of hematocolpometra. AB - Two cases of hematocolpometra, due to imperforate hymen of 15- and 16-year-old girls are presented. The diagnosis was based on computed tomography of the pelvis with contrast given per os, intravenously and per rectum, as well as ultrasound scan of uterus and vagina. PMID- 3052798 TI - The psychological aspects of bone marrow transplant. A staff's perspective. PMID- 3052797 TI - 6p+ and 9q- in two chromosomally distinct clones occurring in a case of myelodysplastic syndrome evolving to acute nonlymphocytic leukemia. AB - Different and unrelated chromosome changes were found to occur in a patient with a myelodysplastic syndrome with rapid evolution to acute nonlymphocytic leukemia. A 6p anomaly was found during the chronic phase and a del(9q) characterized the cells in the leukemic phase. Deletions with a breakpoint in 9q31 appeared to be associated with more aggressive disease. PMID- 3052799 TI - Assessing psychosexual needs of women experiencing lumpectomy. A challenge for research. PMID- 3052800 TI - The position of nursing. Between school medicine and alternative medicine. PMID- 3052801 TI - Liver microsomal inactivation of 4-methyl-5-amino-1-formylisoquinoline thiosemicarbazone as an inhibitor of ribonucleotide reductase. AB - Studies were carried out to determine the effects of preincubation of 4-methyl-5 amino-1-formylisoquinoline thiosemicarbazone (MAIQ) with hepatic microsomes on the ability of MAIQ to inhibit CDP reductase activity in vitro. An aliquot from the 100,000 x g supernatant fraction from this incubation was used in the CDP reductase assay. MAIQ incubated in the absence of microsomes inhibited CDP reductase activity in a dose-dependent manner. At high MAIQ concentration (5 microM) CDP reductase activity was inhibited 95%. When MAIQ (5 microM) was first incubated in the presence of hepatic microsomes and NADPH, CDP reductase activity was inhibited only 30%. This attenuation of MAIQ inhibition was dependent on time of incubation and microsomal protein concentration and showed an obligatory requirement for NADPH or NADH. Significant attenuation was observed at pyridine nucleotide concentrations as low as 0.1 mM. Heat denaturation of microsomal proteins inactivated their ability to attenuate the MAIQ inhibition. Microsomes prepared from Ehrlich tumor cells were ineffective as inactivators of MAIQ. Results of our studies show that hepatic microsomes contain an enzyme(s) which can inactive MAIQ as an inhibitor of CDP reductase. PMID- 3052802 TI - Proteases in cyst fluid from human gross cyst breast disease. AB - Cyst fluid from women with gross cystic breast disease was found to contain protease activity when assayed against [14C]albumin. At least six different proteases were detected when the fluid was fractionated by a combination of S-300 Sephacel, hydroxylapatite, and DEAE-Sephacel chromatographic techniques. The distribution of the proteases appeared to be related to the ionic composition of the fluids. A major protease component, found in both high Na and high K fluids, was isolated. It showed chymotryptic cleavage characteristics against the beta chain of insulin. It was partially inhibited by alpha 2-macroglobulin, N-tosyl-L phenylalanine chloromethyl ketone, and benzamidine but not by leupeptin, pepstatin, N-tosyl-L-lysine chloromethyl ketone, or alpha 1-protease inhibitor. The protease has an apparent molecular weight of 110,000 with Mr 24,000 subunits. This protease may be identical or closely associated with Haagensen's GCDFP-24 progesterone binding protein which was isolated in a similar manner. An imbalance between protease and protease inhibitors in cyst fluid may account for gross cyst formation and may be involved in the tumorigenic process. The accumulation of poorly diffusible peptide fragments, as a result of protease activity, would increase the oncotic pressure leading to enlargement of the cyst cavity as water enters to reestablish osmotic equilibrium. PMID- 3052803 TI - Quantitative studies on the transplantability of murine and human tumors into the brain and subcutaneous tissues of NCr/Sed nude mice. AB - The transplantability of experimental tumors into the brain (i.c.) and s.c. tissues of C3Hf/Sed and athymic NCr/Sed nude mice was examined using quantitative cell transplantation assays. Studies using the immune-competent C3H animals showed that brain is a more favorable site for the transplantation of syngeneic tumor than s.c. tissue and that this is true for nonimmunogenic as well as immunogenic tumors. The capacity of the brain to act as an immunological sanctuary can be overwhelmed by a strong, systemic, secondary immune response such as that evoked by the methylcholanthrene-induced sarcoma FSal. In studies performed using NCr/Sed nude mice, the allogeneic tumor MCaIV was found not to be demonstrably immunogenic. The cell dose required to transplant the tumor into 50% of recipients (TD50) could neither be increased by immunization procedures nor decreased by six Gy whole-body irradiation (WBI) prior to transplantation. Delayed-type hypersensitivity to this tumor was not expressed by nude mice after rechallenge with tumor antigen. The TD50 was again lower for i.c. than s.c. transplantation and the ratio s.c./i.c. was comparable to that found in syngeneic C3Hf/Sed hosts. Three human tumors have been similarly tested. They were: FaDu, a pharyngeal squamous carcinoma; HFSal, a fibrosarcoma; and U87, a malignant glioma. s.c. TD50 values were in all cases significantly higher than those obtained i.c. The ratios TD50 s.c./i.c. ranged from 6.4 to greater than 50 in five studies, substantially higher than those found for transplantation of murine tumors into either the syngeneic or the allogeneic recipients. Six Gy WBI reduced the s.c. TD50 for these tumors, but in each case the value remained significantly higher than that obtained i.c. 19.4 Gy WBI given in 10 equal fractions and followed by i.v. bone marrow rescue reduced further the s.c. TD50 for FaDu. NCr/Sed nude mice demonstrated cross-reacting delayed-type hypersensitivity against FaDu and HFSal. A small proportion of FaDu tumors (less than 2%) displayed a spontaneous halt in growth or even regression. When the host cell infiltrate of these tumors was analyzed, an increase was seen in the proportion of Thy 1.2 and asialo-GM1-positive cells as compared with progressively growing tumors. These data strongly suggest that a residual low level of immune reactivity exists in nude mice against xenotransplanted human tumors. This resistance to s.c. transplantation may be diminished by WBI and is less for intracerebral implantation. PMID- 3052804 TI - Immunocytochemical localization of estrogen receptor in human breast tissue. AB - An immunocytochemical assay for the measurement of estrogen receptor (ER-ICA; Abbott Diagnostics) has been evaluated in 163 human breast carcinomas. Specific binding was observed in the nuclei of 111 of 163 (68%) tumors. An excellent correlation was observed between the ER-ICA and the estrogen receptor enzyme immunoassay. A significant relationship was observed between ER-ICA status and percentage of ER-ICA negative cells, histological grade of malignancy, and mitotic activity of the tumors. A significant correlation was also observed between ER-ICA status and age at mastectomy with 50% of patients with ER-ICA positive breast tumors presenting with their disease over 60 years of age. No association was observed with either tumor size or patient nodal status. Examination of the proportion of negative cells within tumors revealed a trend for the acquisition of poor prognostic features to be associated with an increase in the negative cell population. Data on the recurrence free interval of these patients showed a significant recurrence free advantage in ER-ICA positive patients, particularly those whose tumors contained low numbers of negative cells. PMID- 3052805 TI - Intranucleolar localization of human proliferating cell nucleolar antigen p120. AB - The human proliferation-associated nucleolar antigen p120 was localized to substructures within HeLa cell nucleoli by immunofluorescence and immunoelectron microscopy of cells whose nucleoli were segregated by drug treatment or extracted with nucleases. By indirect immunofluorescence, protein p120 was localized diffusely throughout all interphase nucleoli. However, high resolution immunoelectron microscopy demonstrated that protein p120 staining delineated a network of 20-30-nm diameter beaded fibrils distributed throughout the nucleolus. This distribution was unique compared to that of the nucleolar proteins p145, RNA polymerase I, or B23 which were examined simultaneously. Drug-induced segregation of nucleoli by actinomycin D or dichlorobenzimidazole riboside, followed by immunoelectron microscopy, indicated that protein p120 was concentrated at the periphery of the granular region in segregated nucleoli. In situ nuclease digestion of cells with DNase I and/or RNase A did not release p120 from the nucleolus. Instead, p120 immunoreactivity was retained within phase-dense residual nucleoli. These results provide evidence that protein p120 is associated with, and in fact delineates, a network of fibrils which is retained in the nucleolar residue fraction of proliferating cells. PMID- 3052806 TI - Partial down-regulation of protein kinase C in C3H 10T 1/2 mouse fibroblasts transfected with the human Ha-ras oncogene. AB - Biochemical and immunological comparison of mouse C3H 10T 1/2 fibroblasts and C3H 10T 1/2 fibroblasts transfected with human activated Ha-ras oncogene indicated significantly lower levels of protein kinase C (PKC) activity and protein in the ras-transfected cells. This effect was observed in three clonal cell lines transfected with an activated ras oncogene. Cytosolic extracts of the ras transfected cells contained calcium-activated, phospholipid-dependent protein kinase (PKC) activity at 61% of the level of activity present in C3H 10T 1/2 cells. A similarly decreased level of phorbol ester-binding activity was observed in these cells. Analysis of the subcellular distribution of PKC activity in cells failed to indicate significant differences between these cell lines. Immunoblots showed a lower abundance of the Mr 80,000 PKC in ras-transfected cell homogenates and extracts compared to C3H 10T 1/2 cells. Both C3H 10T 1/2 cells and cells transfected with ras expressed only one of the PKC isozymes as resolved by hydroxylapatite chromatography demonstrating that ras transfection of cells did not induce expression of alternative PKC isozymes. These observations indicate that PKC was partially down-regulated in ras-transfected cells, perhaps resulting from constitutively elevated levels of products of phosphatidylinositol-4,5 bisphosphate hydrolysis. Although C3H 10T 1/2 cells were previously shown to be distinct from NIH 3T3 cells in their sensitivity to transformation by the T24-ras oncogene, ras transformation appears to partially down-regulate PKC in C3H 10T 1/2 cells in a manner identical to that for ras-transformed NIH 3T3 cells. This indicates that down-regulation of PKC directly results from the expression of an activated ras oncogene independently of cellular sensitivity to transformation by expression of ras. The common action of ras transformation and phorbol esters to down-regulate PKC provides a possible mechanism for synergism during multistage carcinogenesis. PMID- 3052807 TI - New monoclonal antibody, 1C5, reactive with human cervical adenocarcinoma of the uterus, with immunodiagnostic potential. AB - A murine monoclonal antibody, 1C5, was produced by fusion of spleen cells obtained from mice immunized with CAC-1, a human cell line of adenocarcinoma derived from uterine cervix, and NS/1 myeloma cells. 1C5 can be used for the staining of routine formalin-fixed and paraffin-embedded tissue sections. 1C5 defined antigen was found to have a molecular weight of 26,000. The 1C5-defined antigen was resistant to neuraminidase and trypsin treatment, but sensitive to periodate treatment, indicating that an epitope of the 1C5-defined antigen is a carbohydrate moiety. Immunohistochemical study using immunoperoxidase staining demonstrated that 1C5 reacted with 87% of adenocarcinomas of the uterine cervix, 39% of endometrial carcinomas of the uterus, 100% of ovarian mucinous cystadenocarcinomas, 43% of ovarian serous cystadenocarcinomas, 45% of adenocarcinomas of the colon, and 40% of gastric adenocarcinomas, thus showing the broad reactivity to adenocarcinoma cells of various origins. However, 1C5 did not show any reactivity to ectocervix epithelium, cervical intraepithelial neoplasia, or squamous cell carcinoma of the uterine cervix. In addition, adenocarcinoma of the uterine cervix exhibited strong cytoplasmic reactivity with 1C5, whereas endometrial carcinoma of the uterus showed the luminal reactivity. 1C5 also reacts with 95% ethanol-fixed malignant cells in cervical smears. PMID- 3052808 TI - Association of DNA content and proliferative activity with clinical outcome in patients with diffuse mixed cell and large cell non-Hodgkin's lymphoma. AB - Formalin-fixed and paraffin-embedded lymph node biopsy specimens from 52 untreated patients with newly diagnosed diffuse large cell (n = 48) or mixed cell (n = 4) non-Hodgkin's lymphoma (NHL) were analyzed for DNA content and proliferative activity (PA) by flow cytometry. The results obtained by flow cytometry were compared with the results of cytogenetic studies performed on 28 of the specimens. The median age of the patients was 65 years (range, 15-84 years) and the male to female ratio was 3 to 2. All patients were uniformly staged and uniformly treated with cyclophosphamide, doxorubicin, procarbazine, bleomycin, vincristine, and prednisone. The flow cytometric results were compared statistically by univariate analysis with the rate and duration of complete remission and survival. Tumors with low PA (greater than or equal to 80% of cells in G0/G1 phase) were found in 65% of the patients; 74% of those with low PA versus only 44% of those with high PA achieved an initial complete remission (P less than 0.02). DNA aneuploidy was detected in tumors of 56% of the patients and was associated with a significantly longer duration of complete remission (P less than 0.01). Both low PA and aneuploidy independently predicted longer survival. The predicted 2-year actuarial survival for patients with tumors with low PA was 68% versus 10% for those with high PA (P less than 0.01). Similarly, the 2-year survival of patients with aneuploid tumors was 60% versus 36% for those with diploid tumors (P less than 0.01). The combination of PA and DNA content categorized the patients into four groups with decreasing 2-year survivals: low PA/aneuploid (n = 20), 77%; low PA/diploid (n = 14), 57%; high PA/aneuploid (n = 9), 32%; high PA/diploid (n = 9), 0%. The flow cytometric results correlated well with those of the cytogenetic studies. We conclude that low PA and DNA aneuploidy, both separately and in combination, predict a favorable clinical outcome for patients with diffuse mixed cell and large cell NHL. PMID- 3052809 TI - Immunohistological and functional analyses of lymphoid infiltrates in human glioblastomas. AB - An immunohistological analysis of tumor tissue obtained from seven patients with malignant gliomas demonstrated varying levels of lymphoid cell infiltration. The tumor infiltrating lymphocytes obtained from each sample were cultured in vitro by a limiting dilution technique. In three of the cases studied many tumor infiltrating lymphocyte microcultures selectively lysed autologous glioblastoma cells but did not lyse allogeneic gliomas, natural killer-resistant fresh melanoma cells or K562 target cells. These cultures were found to consist of CD 3+ cells. In six cases studied a variable number of microcultures lysed both autologous tumor and K562 target cells only. A minority of the microcultures studied were cytolytic for allogeneic glioma cells and fresh melanoma target cells. The cytolytic activity expressed by tumor infiltrating lymphocytes against autologous tumor cells was significantly greater (P less than 0.001) than that obtained by the corresponding peripheral blood lymphocytes cultured in a similar manner. The present immunohistological and functional studies suggest that there is an immune response to human glioblastomas in vivo with an accumulation of cells with antitumor activity at the tumor site. PMID- 3052810 TI - Obesity, genetics, and ponderal set point. AB - A significant proportion of the interindividual variance in human fatness is attributable to genetic factors. This fact is indicated (inter alia) by studies of identical twins demonstrating that the degree of efficiency with which the body uses excess dietary energy for fat storage is, to a considerable extent, inherited. Genetic factors also appear to render particular individuals or groups especially vulnerable to pervasive obesity-promoting influences (excess food and too little exercise) in the environment. If, indeed, much human obesity results from an interaction between a genetically determined disposition to store fat efficiently and a food-laden, overmechanized environment, then preventive and therapeutic strategies should be directed toward (a) helping the susceptible individual protect himself from the "calorie pollution" that surrounds him, and (b) finding pharmacologic agents to modify the inherited metabolic factors that favor excessive fat storage. PMID- 3052811 TI - Permanent administration of d-fenfluramine in rats: paradoxical effects. AB - In order to test the hypothesis of Levitzky that d-fenfluramine (d-F) acts by modifying the ponderal set-point, we compared the effects of a permanent infusion of d-F on food intake and body weight (BW). The effect on the weight persisted as long as the infusion; the clear-cut anorectic effect lasted only a few days. This paradox is compatible with the set-point hypothesis. In rats rendered overweight by insulin treatment, the d-F-induced decrease in BW was approximately four times smaller than in controls. In rats rendered overweight by a cafeteria diet, the decrease in BW was twice as large in permanently cafeteria fed rats as in cafeteria, then, ad lib fed rats. In rats rendered underweight by a restricted chow diet and then returned to an ad lib feeding, the final BW depended only on the doses of d-F (0.6 or 12 mg/kg BW/day), whatever the weight at the beginning of infusion. Thus, the underweight paradigm fits well with the set-point hypothesis; the overweight paradigm fits only partially. PMID- 3052812 TI - Carbohydrate cravings: a disorder of food intake and mood. AB - Current findings on the relationship between excessive appetite for carbohydrate rich foods and mood disorders may explain repetitive weight gain or the inability to lose weight among some obese individuals. Obese individuals who crave carbohydrates, exhibit positive changes in mood after carbohydrate intake; individuals suffering from Seasonal Affective Disorder experience a craving for carbohydrate-rich foods in association with their mood disturbances. Brain serotonin may be involved in these disorders of affect and appetite; thus therapies that mimic the effect of carbohydrate intake on the synthesis and release of this neurotransmitter may be useful in treating obesity arising from these causes. PMID- 3052813 TI - A controlled trial of d-fenfluramine in bulimia nervosa. AB - A double-blind, placebo-controlled trial of d-fenfluramine in bulimia nervosa was undertaken in order to assess its efficacy in controlling bulimic behavior and relieving more general symptoms. A high proportion of the patients evaluated were reluctant to enter the drug trial in spite of the offer of additional supportive psychotherapy and counselling on dietary control. Moreover, 17 out of the 42 enrolled patients withdrew halfway through the 12 week trial. Were it not for this high rate of defaulting, there might be clearer support for the efficacy of d-fenfluramine in reducing the frequency of overeating and self-induced vomiting in these bulimic patients. An unexpected finding was that among the noncompleters, those on d-fenfluramine had experienced relief of their bulimic symptoms. The persistence of depressive symptoms and features of the eating disorder probably contributed to the noncompleters leaving the trial. Reassuring findings were the absence of weight loss and serious unwanted effects from d fenfluramine. By itself, d-fenfluramine did not benefit some of the patients with severe bulimia nervosa, but it may yet prove a useful adjunct to psychological treatments. PMID- 3052814 TI - Factors that may effect the reduction of hunger and body weight following d fenfluramine administration. AB - Three studies have been undertaken to investigate why there are individual differences in the response to d-fenfluramine with respect to food intake and hunger in the short term and on body weight loss in the long term. Fenfluramine and norfenfluramine plasma levels have been used as probes to help detect and normalize these variances. In a single dose ranging volunteer study (0, 30, 40, and 60 mg), d-fenfluramine levels were significantly related to caloric intake and hunger rating scales when compared individually, and the slopes of the regression lines showed intersubject variation. These slopes, an index of each subject's response to fenfluramine, appear to be related to both the percentage underweight and more weakly to the percentage overweight. Those subjects at the extremes of weight showed a greater response to a given drug level. In two placebo-controlled 3 month studies (30 mg/day), the variances in weight loss were not explained by steady state drug levels, the percentage overweight, initial weight, duration of obesity, or caloric intake even when weight loss was normalized for differences in drug levels. Age, however, was significantly related to weight loss, with each additional 10 years increasing weight loss by approximately 1 kg. If confirmed, the sensitivity of fenfluramine anorexia may be an objective acute test of the central control of food intake. However, in long term clinical studies, drug levels were only weakly related to weight loss and other undefined factors seem to determine which patients responded better to fenfluramine treatment. PMID- 3052815 TI - Efficacy and safety of dexfenfluramine in obese patients: a multicenter study. AB - We have evaluated the effects of dextrofenfluramine treatment on body weight control during a 90 day period, in obese patients on a calorie-restricted diet. The weight loss in dextrofenfluramine-treated patients was significantly higher than in placebo group. The rate of weight loss was linear up to the end of the trial in d-fenfluramine patients. Neural disturbances (vertigo, headache, depression) were the most frequent side effects observed in both the d fenfluramine and in the placebo-treated groups, without significant differences between the groups. A total number of 23 patients in the dextrofenfluramine group and 20 patients in the placebo group complained side effects. Six patients (five in the d-fenfluramine group and one in the placebo group) discontinued the treatment, due to the side effects. No modifications of the biochemical parameters considered (fasting blood glucose, bilirubin, alkaline phosphatase, creatinine, blood cell counts, asparate-amino transferase (AST), alanine-amino transferase (ALT), total plasma and HDL cholesterol, and triglycerides) were observed at the end of the trial. A significant reduction of total serum cholesterol was observed in both groups at the end of the period of treatment. In conclusion, dextrafenfluramine was proved to be in short term trials an effective and safe tool in overweight control in obese patients. PMID- 3052816 TI - Effect of 6 months therapy with dexfenfluramine in obese patients: studies in the United Kingdom. AB - The efficacy of dexfenfluramine in inducing weight loss and its clinical acceptability were assessed in two studies carried out in a hospital obesity clinic and general practitioner's office. Seventeen patients who had gained an average of 0.7 +/- 1.9 kg during 12 weeks treatment with diet and placebo lost a mean of 2.9 +/- 0.5 kg after 12 weeks sequential treatment with dexfenfluramine 15 mg twice daily (p less than 0.001). In a second trial, 29 patients treated for 24 weeks with dexfenfluramine showed significantly increased, and continuing, weight loss after 24 weeks (7.0 +/- 0.8 kg) compared to that after 12 weeks of treatment (5.7 +/- 0.1 kg; p = 0.05). The incidence of side effects in both trials was lower than that reported in previous studies of racemic dl fenfluramine. The clinically significant weight loss and low incidence of unwanted effects suggest that dexfenfluramine has a role in the treatment of refractory obesity. PMID- 3052817 TI - Effects of their nutrient precursors on the synthesis and release of serotonin, the catecholamines, and acetylcholine: implications for behavioral disorders. AB - Authentic foods affect brain serotonin synthesis by modifying brain tryptophan levels, carbohydrates increasing and proteins decreasing these levels. The carbohydrate-induced rise in brain serotonin tends to diminish the likelihood that one carbohydrate-rich, protein-poor meal or snack will be followed by another. This mechanism is apparently disturbed in carbohydrate-craving obesity, which may explain why this syndrome responds well to d-fenfluramine, a serotoninergic drug. Pure nutrients like tyrosine or choline can also affect the rates at which their neurotransmitter products, the catecholamines and acetylcholine, are synthesized in and released from nerve terminals, suggesting that these compounds may find uses as drugs. PMID- 3052818 TI - Psychotropic drug induced weight gain: mechanisms and management. AB - Most of the drugs commonly used in the treatment and prophylaxis of depression, mania, and psychotic illness have, as one of their prominent side effects, the ability to increase appetite, stimulate carbohydrate craving, and promote weight gain. These side effects are troublesome to patients, and frequently constitute a major reason for premature discontinuation of therapy. This review examines the relative likelihood of the occurrence of appetite stimulation and weight gain with various psychotropic medications. Potential mechanisms of these effects and strategies to minimize or avoid weight gain during pharmacotherapy of psychiatric illness are examined. Evidence suggests that those compounds, which either antagonize or downregulate serotonin receptors, are more likely to stimulate carbohydrate hunger and weight gain. Amitriptyline, chlorpromazine, mesoridazine, thioridazine, and lithium are most likely to produce weight gain. Compounds that have more pronounced serotonergic action, such as fluoxetine and fenfluramine, are more likely to decrease carbohydrate craving and promote weight loss. PMID- 3052820 TI - Place of dexfenfluramine in the management of obesity. AB - The treatment of obesity remains a puzzling challenge because long-term maintenance of weight loss--one of the most suitable goals--is rarely achieved with conventional methods. Among the theoretical measures able to maintain a permanent and bearable constraint to obtain the maintenance of weight loss is long-term (life-long?) pharmacotherapy. Dexfenfluramine is a good candidate for such a use; the demonstration of its long-term efficacy, i.e., maintenance of the initial weight loss with no escape with time, is in progress (ISIS study). Although the correct use of dexfenfluramine in obese patients remains to be more precisely defined, its pharmacological profile leads us to consider that it could be useful for those with normo or hyperphagia and stress induced compulsive feeding behavior, and/or for avoiding relapse in restrained patients. The use of dexfenfluramine must not excuse other types of interventions when needed. PMID- 3052819 TI - The effect of altered tryptophan levels on mood and behavior in normal human males. AB - The effect of lowering or raising tryptophan levels was studied in normal males using amino acid mixtures that were tryptophan free or tryptophan supplemented. Tryptophan depletion caused a small but significant alteration in food selection in subjects allowed to select from a buffet. Although carbohydrate and total kilocalories selected were unchanged, significantly less protein was chosen. Tryptophan depletion also caused an acute lowering of mood, suggesting that low serotonin (5-HT) may be involved in the etiology of clinical depression in some patients. No effect of altered tryptophan levels was seen in a laboratory test of aggression. However, a study on vervet monkeys indicated that altered tryptophan levels can influence aggression when the animals are at a high level of arousal. High arousal is known to increase the firing rate of 5-HT neurons. In a preliminary study, tryptophan had a therapeutic effect in aggressive schizophrenic patients. The best effect was seen in impulsive patients, which may have been related to high arousal in these subjects. Although carbohydrate is capable of raising brain tryptophan, not all carbohydrate-induced behaviors are mediated by tryptophan. Thus, sucrose was capable of attenuating alcohol intoxication in normal human males without altering blood alcohol concentrations. However, tryptophan had no effect on ethanol intoxication. The challenge for the future is to define the conditions under which alterations in tryptophan levels can influence brain function. PMID- 3052821 TI - Dexfenfluramine: effects on food intake in various animal models. AB - The effects of dexfenfluramine on food intake of rats are compared and contrasted among several paradigms. Paradigms that involve overfeeding rather than deprivation, show a greater effect of the agent. Furthermore, both stress-induced eating as well as a food-motivated response (running) are particularly sensitive to inhibition by dexfenfluramine. The results of chronic administration of dexfenfluramine on both behavior and neurochemistry are reviewed, with particular reference to brain 5-HT. The effects of dexfenfluramine on diet selection and conditioned preferences are discussed. Finally, we suggest that species differences may exist in either the neurochemical and/or behavioral responses to dexfenfluramine. PMID- 3052822 TI - Dexfenfluramine and feeding reward. AB - This review describes background experiments and new findings showing that dexfenfluramine inhibits self-stimulation of a lateral hypothalamic (LH) feeding reward system. Earlier work suggested that self-stimulation excites a pathway to the ventral tegmental area (VTA), where the mesolimbic dopamine system is involved in self-administration of food and drugs. LH stimulation also excites taste neurons in the nucleus tractus solitarius (NTS). This helps explain why stimulation can induce feeding responses and also reward self-stimulation responses. Stimulation-induced feeding and self-stimulation are modulated by physiological signals that control an animal's appetite and body weight. An animal will self-stimulate at a slower rate after it has just eaten, or if it is overweight, or if it is given an anorectic dose of insulin. At the same time, responses to turn off automatic stimulation tend to increase. Paradoxically, racemic fenfluramine decreased both self-stimulation and stimulation-escape, which suggested overall lethargy, but new results show this can be avoided by using just the d isomer. Dexfenfluramine inhibited LH self-stimulation but not stimulation-escape. It also released serotonin in the LH, as shown by microdialysis. These results suggest that dexfenfluramine can release serotonin that has as one of its effects the inhibition of circuitry for feeding-reward. PMID- 3052824 TI - Serotonergic mechanisms of depression. AB - Data gathered for the past 30 years suggest 5-HT abnormalities in depression, but it is still unknown whether serotonin has a major causal role. Depression is often regarded as a result of defective serotonergic function, but it is also possible that this defect is an indication of an ineffective compensatory response to abnormally high serotonergic function. Abnormal 5-HT function could be a result of disturbances at various pre- and postsynaptic levels. The disturbances reported at most of these levels are largely subject to dispute. Specific abnormalities can occur in specific subgroups and can be either state or trait dependent. Nevertheless, 5-HT hypotheses have been helpful for the development of drugs, although even here there is disagreement as to their actual mechanism of action. Two recent developments can help to clear up these uncertainties. First, the discovery of many types of 5-HT receptors and the increasing availability of drugs specifically related to them provides new adaptable research tools as well as the possibility of new therapies. Second, data provided by studies of humans and by animal models suggest that vulnerability to depression depends on disturbances of both 5-HT and the hypothalamohypophysial axis. PMID- 3052823 TI - Progress in assessing the role of serotonin in the control of food intake. AB - There is evidence that serotonin inhibits food intake, particularly intake of carbohydrate and that induced by activation of catecholamine-containing neurons in different brain circuits. An agent that has contributed considerably to the hypothesis of a role of serotonin in feeding is fenfluramine, used as an anorexigenic drug in obese people. Experiments using synaptosomal preparations for studying monoamine uptake and release have shown that d-norfenfluramine preferentially releases serotonin from a reserpine-insensitive compartment. Studies on brain monoamine release and metabolism in intact animals have shown that d and l isomers of fenfluramine at relatively low doses have a specific action on brain serotonin and catecholamines, respectively. Several findings suggest that d-fenfluramine and d-norfenfluramine cause anorexia by increasing the availability of serotonin at postsynaptic receptors. Evidence has recently been provided that d-fenfluramine uses preferentially serotonin1 sites, particularly of the serotonin1B type, in the rat brain to cause anorexia in this animal species. Activation of serotonin1A sites by agents such as 8-OH-DPAT and buspirone instead has been shown to cause overeating. It is suggested that serotonin1B sites in the hypothalamus and serotonin1A sites in the serotonin neurons of the midbrain raphe nuclei mediate these effects. Evidence is provided that [3H]d-fenfluramine binding to rat brain membranes is different from serotonin uptake sites ([3H]imipramine binding) and serotonin receptors. It is, however, displaced by some drugs using serotonin to cause anorexia, raising the possibility that it is somewhat related to serotonin mechanisms involved in feeding control. These studies provide evidence that the serotoninergic system in the brain is a likely target for drugs affecting food intake and suggest new ways to develop novel and potent strategies for the treatment of clinical hyperphagia and anorexia. PMID- 3052825 TI - Neurochemical profile of tianeptine, a new antidepressant drug. AB - Tianeptine is a new effective antidepressant drug. However, its neurochemical profile in animals differs from that of tricyclic or atypical antidepressants. In the present study, we compared the ex vivo effects of tianeptine on platelet serotonin uptake to those of clomipramine. Ex vivo, after subchronic (15 days, washout 24 h) treatment (10 mg/kg/day i.p.) in rats, tianeptine induced an increase (30%) in [14C]serotonin uptake at a [14C]serotonin concentration of 500 nM while clomipramine induced a decrease (40%) in [14C]serotonin uptake. Stimulation of uptake affected mainly Vmax but not Km. Tianeptine did not inhibit monoamine oxidase (MAO), MAOA or MAOB activity. In vitro, there was no binding of tianeptine to any of the various receptors examined: alpha- and beta-adrenergic, dopamine, serotonin, imipramine, GABA, glutamate, benzodiazepine, muscarinic, histamine, Ca2+ channels. After chronic administration (10 mg/kg/day for 15 days) tianeptine did not alter the concentration (Bmax) or the affinity (Kd) of alpha 2, beta-1, serotonin-1, serotonin-2, imipramine, benzodiazepine, or GABAB receptors. The major effect of tianeptine in rat platelets and synaptosomes is a small increase in 5-HT uptake after subchronic administration. PMID- 3052826 TI - Role of drug therapies in the treatment of alcoholism: alcohol and anxiety- alcohol and depression. AB - Drug therapies are often used in therapeutic programs for alcoholics. This paper focuses on the use of psychotropic agents which can be used at different stages in the alcoholic disease process. It does not cover specific drug therapies for alcoholism (aversion, chemical restraint, dissuasion), nor nonspecific drug therapies (vitamins, magnesium) the interest and limits of which are well known. All types of psychotropic drugs, antianxiety drugs, antidepressants, thymoregulators, neuroleptics, and beta-blockers are used, with two main indications: (a) to prevent or to treat the withdrawal syndrome and (b) to diminish the psychopathological factors related to alcoholism (anxiety or depression, for example), in order to avoid pathological recourse to alcohol. Recently, attempts have been made to decrease the craving for alcohol in humans after detoxification. Although psychotropic drugs are often used in the management of alcoholics, such management is not limited to them, but also includes psychotherapy and social measures. PMID- 3052827 TI - Position of tianeptine among antidepressive chemotherapies. AB - The advent of a new antidepressant is always received with both interest and scepticism by clinicians. No new compound has yet been shown to be more efficient than imipramine in the treatment of depression. In determining the position of a compound among the antidepressants, four levels are to be considered: (a) Chemical family: Tianeptine is a tricyclic compound of dibenzothiazepine type. It is the only representative of this subgroup. (b) Biochemical activities: Tianeptine increases the presynaptic uptake of serotonin after single as well as repeated administration; but this action is not linked to any effect on the 5-HT post-synaptic systems. Electrophysiological modifications are observed in the locus coeruleus (noradrenergic) at 2.5 times higher doses; nevertheless, the essential action involves the serotoninergic systems. Tianeptine has no affinity for alpha 1 adrenergic and H1 antihistaminic receptors; this affinity is responsible in a large part for the sedative anxiolytic properties of antidepressants. Tianeptine has no affinity for the muscarinic receptors. It has secured its place among the non-anticholinergic antidepressants. The mechanism of action or the therapeutic profile cannot be inferred from this original biochemical profile. (c) In clinical trials, the double-blind trials confirm the antidepressant efficacy in essentially neurotic depressive syndromes, similar to that of imipramine, nomifensine, amitriptyline. They do not show a shorter onset time of antidepressant activity. The therapeutic profile appears to be neither stimulating nor sedative. Some arguments are in favor of an anxiolytic activity. Others favor a more psychotonic profile. Tianeptine can be classified among the mid-position antidepressants. (d) With respect to clinical acceptability, tianeptine has no anticholinergic effect and no cardiovascular effect. PMID- 3052828 TI - A morphological analysis of an in vitro model of cytotoxic antitumor activity. AB - Models of adoptive immunotherapy and cytotoxicity of lymphocytes isolated from tumor tissues were studied. A microcytotoxicity model utilizing serum-free culture conditions was evaluated by light microscopy, scanning electron microscopy, and transmission electron microscopy for antitumor activity. By examining morphologically the relationship between peripheral blood mononuclear cells and tumor-infiltrating lymphocytes, it was demonstrated that the latter and macrophages had morphological features similar to the cytotoxic cells obtained in vitro from peripheral blood. When allowed sufficient time in our culture conditions, the tumor-infiltrating cytotoxic cells seem to kill the tumor cells within a few days to many months. The ultrastructural morphology of the interaction between the cytotoxic cells and the tumor cells was described as well as some proteinaceous secretory granules that seem to be transferred to the tumor cells through these interactions. PMID- 3052829 TI - Correlation between tumor proliferation and serum levels of beta 2-microglobulin and thymidine kinase in malignant lymphomas. AB - We have studied the serum beta 2-microglobulin (beta 2m) and thymidine kinase (TK) levels in 19 newly diagnosed lymphoma patients. The proportion of the S phase cells (SPF) was determined by flow cytometry from subsequently taken tumor biopsy material. A positive correlation between SPF and TK (r = 0.4, P = 0.1), but not between SPF and beta 2m, was seen in the whole material. Sixty-three percent of the high-grade malignancy non-Hodgkin lymphomas (5/8) showed high proliferative activity in both the SPF and TK analyses. Furthermore, high tumor SPF and enhanced serum TK levels reflected equally well and consistently the clinical outcome of the underlying disease. PMID- 3052830 TI - AIDS: a global problem. AB - AIDS has emerged as one of the foremost infectious disease epidemics of the twentieth century. This review summarizes the magnitude of the AIDS problem and the frequency of infection with the causative agent, the human immunodeficiency virus (HIV). The natural history of HIV infection suggests that a high proportion of persons still healthy but infected with HIV will become immunosuppressed over the next decade. These persons are capable of transmitting HIV to others as a venereal disease as well as by blood and blood products. Thus, the epidemic will grow not only because more cases will appear among persons already infected but also because new infections will occur. However, the incidence of new HIV infections within the homosexual community is already slowing because of changing behavior after the advent of AIDS. The spread of HIV infection within the heterosexual community will probably never be as extensive as that in the homosexual community during the early 1980s because of different behavior patterns. Containment of the epidemic will require an effective vaccine as well as changes in personal life style, but it is unlikely these developments will eradicate HIV infection as a public health problem. PMID- 3052831 TI - Mechanisms of immune suppression in AIDS: possibilities of prevention. AB - AIDS is primarily a disease of the CD4-expressing helper-inducer T lymphocytes and is caused by human immunodeficiency virus (HIV), a retrovirus of the lentivirus subgroup. The interaction of HIV envelope glycoprotein (gp 110) with the CD4 molecule itself accounts for the virus selective tropism and for its pathogenicity, which result in the ultimate destruction of immunocompetent cells. Viral replication occurs preferentially in case of activation of the infected target cells, which explains why multiple infections, frequent in at-risk subjects, might play the role of cofactors and enhance HIV dissemination. There is yet no proof that an effective immune response might be mounted against this virus. Based on the pathophysiological model that can be drawn from HIV biological properties, many therapeutic schemes can be proposed to interfere with the complex pathway of interaction between host and virus. Nevertheless, the use of retroviral agents appears to be central to all these new strategies, since disease progression seems indeed directly related to the extent of viral replication. PMID- 3052832 TI - Mechanisms of HIV-associated immunosuppression. AB - HIV antigens were identified in PBL obtained from HIV-positive patients, using IF and IEM. Studies of the phenotype of HIV-containing lymphocytes showed that OKT4+ cells were the principal target of the virus. Approximately 5% of infected cells were multinucleated. Almost all infected and about 30% of uninfected PBL displayed Ab-C3 complexes on the cell surfaces. Sera from HIV-positive patients contained Ab reacting with cell membranes and intracellular structures of PBL from normal subjects, as demonstrated by IEM. The presence of Ig-C3 complexes on a high percentage of HIV-positive- or negative-PBL suggests that the Ab-C mediated lympholysis may represent a major mechanism of lymphatic tissue destruction in HIV-infected patients. PMID- 3052833 TI - Use of monoclonal antibodies in immunocytochemical localization of estrogen receptors in ovarian cancer. AB - Ovarian cancer specimens from 45 patients were assayed for the presence of estrogen receptor (ER) utilizing highly specific monoclonal antiestrophilin antibodies and the peroxidase-antiperoxidase technique. Results were compared with the conventional ER determination of the dextran charcoal method (DCC) and were in concordance in 91% of cases. This technique was also used to analyze ER in fine needle aspiration biopsy specimen from four patients with metastatic ovarian tumor. These results suggest that antireceptor monoclonal antibody in immunohistochemical analysis is an effective tool in the evaluation of estrogen receptor content in ovarian cancer. This technique can be easily performed at community hospitals and may be extended to the evaluation of fine needle aspiration biopsies and to analyze specimens of insufficient size for biochemical assay. PMID- 3052834 TI - Clinical applications of carcinoembryonic antigen. AB - Although carcinoembryonic antigen (CEA) has been the subject of interest for many investigators for over 20 years, many questions still remain unanswered concerning the CEA molecule. These include the ultimate clinical potential of CEA as a tumor marker (specificity, sensitivity, distribution), the biological role of CEA, and the genetic control of CEA synthesis. Initially, much of the work with CEA concerned its physicochemical and immunochemical properties, as well as of cross-reacting molecules, and methods of serologic detection of CEA. More recent studies have focused on cloning of the CEA gene and genetic control of CEA production. An extensive literature exists concerning the role of serum CEA assays and their potential value in determining the prognosis and monitoring of patients affected with cancers of various organs. Despite extensive research into the biology of CEA, few papers deal with the application of CEA immunohistochemistry and immunocytochemistry as concerns normal cellular development, the degree of tumor anaplasia, and the various diagnostic problems of surgical pathology. There has also been a great deal of interest in the utility of both polyclonal and monoclonal antibodies to CEA both in the radioimmunolocalization and potential therapy of CEA-producing tumors. This review summarizes the past and current findings of the clinical applicability of the serum measurement of CEA and examines the status of radioimmunolocalization of tumors as a basis for effective antibody targeted immunotherapy in the future. PMID- 3052835 TI - Analysis of ras gene expression in stomach cancer by anti-ras p21 monoclonal antibodies. AB - Using anti-ras p21 monoclonal antibodies, RASK-3, which reacts with all of Ki-, N , and Ha-ras p21, we examined by immunohistochemistry the expression of p21 in human gastric cancer (80 cases) and benign gastric lesions (32 cases). Ten percent formalin fixed tissues were studied. Ras p21 was positive in 51 cases (64%) out of 80 cases and partially positive in 12 cases (15%) at the cancerous areas. Ras p21 was partially positive in 7 cases (9%) at the noncancer areas of the same slides. Intestinal metaplasia and normal parietal cells were also often positive. In the study of 32 cases of benign stomach lesions, 2 out of 3 cases of atypical hyperplasia (ATP) and 3 out of 11 cases of stomach ulcer with regenerating epithelials were positive. Ras p21 was more dominantly expressed in the well-differentiated type of stomach cancer than the poorly differentiated type. Expression of ras p21, however, was not correlated either with the grades of cancer invasion or with the types of cancer infiltration. PMID- 3052836 TI - Detection of transforming sequences in human melanomas and leukemias. AB - DNA extracted from fresh solid human melanoma tumors and untreated acute myeloid leukemic cells was used in two assays designed to detect oncogenes, based on the transfection of murine NIH 3T3 fibroblasts followed by selection of transformed cells in low serum concentration or induction of tumors in athymic mice. Ras and non-ras oncogenes were detected. PMID- 3052837 TI - Secondary neoplasms as a consequence of transplantation and cancer therapy. AB - A complication of various therapies is an increased incidence of cancers. We present data on 3117 types of cancer that developed in 2915 immunosuppressed organ-transplant recipients. The predominant tumors are lymphomas, skin and lip carcinomas, vulvar and perineal carcinomas, in situ uterine-cervical carcinomas, and Kaposi's sarcoma (KS). Tumors appear a relatively short time after transplantation, the earliest being KS at an average of 23 months and the latest vulvar and perineal cancers presenting an average of 98 months after transplantation. Cytotoxic drugs given to cancer patients may cause secondary neoplasms either by a direct carcinogenic effect or, indirectly, through depression of immunity. The most common secondary malignancies are leukemias, lymphomas, and bladder carcinomas. Ionizing radiation causes cancer, either by a direct carcinogenic effect on cells in the radiation field, or indirectly by depressing immunity. The most common malignancies are leukemias and bone sarcomas. PMID- 3052838 TI - Thymopentin treatment in genital warts of long duration. AB - The immunomodulatory effect of thymopentin as therapy in genital warts of long duration and the proliferative responses of the patient's peripheral blood mononuclear cells were investigated in this pilot study. The observations in 6 patients suggest that subcutaneous injections of thymopentin (50 mg) beneficially influence the systems of therapy-resistant genital warts. The small number of patients and controls used in the assessment of the proliferation responses allows only a descriptive analysis of the results, but definitive trends can be observed in the Con A- and PHA-induced proliferation tests. These clinical observations, the theoretical considerations, and the low rate of side effects of thymopentin reported also by other investigators all emphasize the importance of further well-controlled double-blind studies on the treatment of genital warts with thymopentin. PMID- 3052839 TI - Biological response modifiers in cancer therapeutics: potentialities and limitations. AB - With increased knowledge of the mechanisms of host defenses against tumors, both immunological and nonimmunological in nature, and of the relationships between tumors and host cells, it becomes possible to develop agents aimed at modifying those relationships to therapeutic advantage. The task is facilitated by the advent of recombinant DNA technology, which makes it possible to produce biologicals in quantity and purity adequate for studies in vivo. The potentialities of biological response modifiers (BRMs) are primarily due to their specificity and/or selectivity of action, whether these attributes are related to receptors/antigens present on neoplastic cells or on cells of the immunological regulatory networks or to nonspecific activation of host defenses. The limitations are related to the fact that in certain cases treatments are too specific for a tumor cell within a heterogeneous cell population; these agents also show toxicities that, albeit usually unrelated to their antitumor action, can in some cases be severe. PMID- 3052840 TI - Problems and strategies for bone marrow transplantation in acute leukemia and chronic myelogenous leukemia. AB - Certain marrow transplant protocols can now result in a 50-70% long disease-free survival and low relapse rates in acute leukemia (AL) in CR1, CR2, or CML following cytoreduction and HLA-identical marrow infusion. Two-thirds of deaths are due to acute and chronic graft-versus-host disease (GVHD) or viral infection. The other deaths are due to toxicities of the cytoreductive treatment. Prevention of GVHD has been tried by treatment after the transplant or treating the marrow (lymphocyte depletion). Cyclosporine (CsA) or CsA plus methotrexate has reduced acute GVHD but not chronic GVHD. Marrow has been treated with monoclonal antibodies and lectins or elutriated to decrease numbers of T lymphocytes. Some studies have been effective, but the majority have shown an increased number of rejections or leukemic relapses. Apart from teratogenic effects, thalidomide has minimal toxicity. It effectively prevents and treats acute and chronic GVHD in rodent models. Clinical trials will soon begin. Mismatched related or matched unrelated donors have been employed in the clinic with limited success. Alternatively, autologous transplantation in acute leukemia has shown promising results. Possible solutions to remaining problems and strategies will be discussed. PMID- 3052841 TI - Leukaemia and lymphoma treatment with autologous bone marrow transplantation: preclinical studies. AB - Following the limited success of T-cell depletion in patients undergoing allogeneic bone marrow transplantation (BMT), recent interest is focused on autologous BMT. The selection of sensitive methods for detecting residual lymphoid malignancy, the choice of very efficient complement-fixing lytic monoclonal antibodies against residual disease, and the "tailoring" of these antibodies (or their cocktails) to individual patients lead to a very high remission rate in acute lymphoid leukaemia (ALL) of bad prognosis and second remission. The future of this extension of chemotherapy toward BMT seems to be promising, but controlled trials are required to show that the purging of bone marrow contributes to this good result. Very efficient antibodies for elimination of B lymphoma cells are also available. PMID- 3052842 TI - Management of bacterial and fungal infections in bone marrow transplant recipients and other granulocytopenic patients. AB - A review of studies on the effect of different types of gastrointestinal decontamination and protective environment on infectious complications in granulocytopenic patients is given, and the effect of these measures on graft versus-host disease after allogeneic bone marrow transplantation is discussed. It is concluded that complete gastrointestinal decontamination of patients nursed under conditions of strict reverse isolation will maximally prevent infections, graft-versus-host disease, and lung complications and therefore is the treatment to be preferred for patients undergoing bone marrow transplantation. Since selective decontamination is as effective in preventing bacterial and fungal infections as is complete decontamination, this treatment is to be preferred for other patients with a greatly reduced resistance to these infections. The reason is that, for this type of patient, selective decontamination can be performed without the use of strict isolation facilities and in this way is less laborious and less of a burden for the patient. Besides this, the number of patients that can be treated will not be limited by the number of available facilities for strict reverse isolation, which can be reserved for bone marrow transplant patients. PMID- 3052843 TI - Plasmacellular hyperplasia after allogeneic bone marrow transplantation. AB - B lymphoid cells with monotypic immunoglobulin (Ig) expression, i.e., a single Ig heavy chain isotype combined with a single light chain type, were observed in five of nine patients after allogeneic bone marrow transplantation. Three patients exhibited plasmacellular hyperplasia of lymph nodes and spleen; in one case the monotypic cells were immunoblasts in a lymph node, and one patient suffered from an immunoblastic lymphoma along the gastrointestinal tract. We present the clinical history and the detailed (immuno) histologic analysis of lymphoid tissue from three patients. In DNA analysis of lymph node using DNA probes specific for the JH-gene segment of Ig heavy chains and for light chains, bands indicative of single Ig gene rearrangements were not observed. Thus, the monotypic B lymphoid elements represent polyclonal B cell expansion. PMID- 3052844 TI - Mitogenic, immunoadjuvancy, and genetic studies on fatty acyl maltose. AB - Three synthetic glycolipids, maltose tetrapalmitate (MTP), maltose hexastearate (MHS), and maltose hexalinoleate (MHL) prepared as nontoxic lipid A analogs, and Escherichia coli lipopolysaccharide (LPS) were assayed for their mitogenic activity using spleen lymphocytes in nine inbred mouse strains and three F1 hybrids. The MTP and LPS were also assayed for their ability to enhance plaque forming cell (PFC) responses using sheep red blood cells as the antigen in the same inbred mouse strains and F1 hybrids, The mitogenic activity of synthetic glyco-lipids was several fold lower than that of LPS and MHL was inferior to MTP and MHS. DBA/2J was the most responsive strain for MTP and DBA/1J and C3H/HeJ the least. The mitogenic activity of MTP was generally in agreement with the PFC response stimulation by it. Low-dose cyclophosphamide treatment of mice synergized MTP for PFC response augmentation. Genetic studies on MTP mitogenicity revealed that 90% of responder DBA/2J X nonresponder C3H/HeJ F1 hybrids had intermediate mitogenic activity. Among F2, 73% had intermediate-high activity and 27% were nonmitogenic. Among F1 X C3H/HeJ backcrosses 11% had high, 56% intermediate, and 33% had no mitogenic activity, whereas, for the F1 X DBA/2J backcross, 14% had high, 36% intermediate, and 50% low or negligible activity. The data favored a single gene for MTP activation of immune cells. PMID- 3052846 TI - [Physicians, pharmacists and Czech archaeology]. PMID- 3052845 TI - Bactericidal efficacy of metronidazole against bacteria of human carious dentin in vitro. AB - The bactericidal efficacy of metronidazole against bacteria in carious dentin was clarified by measuring (1) the difference between bacterial recovery from suspensions of carious dentin on metronidazole-containing BHI-Blood agar plates (10 micrograms/ml) and control plates and (2) the difference between bacterial recovery from carious lesions of freshly extracted teeth, covered by alpha tricalcium phosphate (TCP) cement containing metronidazole (5%) for 1-3 days and that covered by TCP only. More than 10(3) bacteria per milligram sample were recovered from carious dentin. More than 99% of the bacteria were, however, not recovered when samples were inoculated on metronidazole-containing BHI-Blood agar plates or when the lesions were covered by TCP cement containing metronidazole, indicating that metronidazole effectively disinfected the carious dentin. PMID- 3052847 TI - [Do polycystic kidneys decrease in size in patients on the dialysis transplantation program?]. PMID- 3052848 TI - [The most common diseases among the population of Prague and surrounding area in the 14th century according to the Vetularius of Albik of Unicov]. PMID- 3052849 TI - Cytoskeletal organization and collagen orientation in the fish scales. AB - Immunofluorescence and electron microscopy were used to analyze the relationships between the organization of collagen fibrils in elasmoid scales, and the orientation of microtubules and actin microfilaments in the scleroblasts producing this collagenous stroma. Attention was focused on the basal plate of the scales because of the highly ordered three-dimensional arrangement of the collagen fibrils in superimposed plies forming an acellular plywood-like structure. The collagen fibrils are synthesized by the scleroblasts forming a monolayered pseudo-epithelium, the hyposquama, at the lowest surface of the scale. Fully developed scales with a low collagen deposition rate were compared with regenerating scales active in fibrillogenesis. When an ordered array of the collagen fibrils is found, the innermost collagen fibrils are coaligned with microtubules and actin microfilaments. Thus, because of this coalignment, microtubules and actin microfilaments of the hyposquamal scleroblasts are subjected to consecutive alterations during the formation of the plies of the basal plate. The sequence of events when the collagen fibrils change their direction from one ply to the other in the basal plate is deduced from immunofluorescence and phase-contrast-microscopic observations. During the formation of the orthogonal plywood-like structure in the regenerating scales, first microtubules may change their curse with a rotating angle of about 90 degrees; then, actin microfilaments are disorganized and reorganized by interacting mechanically with the microtubules with which they are coaligned. Collagen fibrils are synthesized in a direction that is roughly perpendicular to that of the preceding ply. The unknown signals inducing the change in direction of the cytoskeleton may be transmitted throughout the hyposquama via gap junctions. PMID- 3052850 TI - Flunarizine (Sibelium) in the prophylaxis of migraine. An open, long-term, multicenter trial. AB - Sixty-four Spanish neurological centers participated in a study designed to evaluate the efficacy and safety of flunarizine in migraine. One thousand four hundred and thirty-five outpatients (367 [25.6%] males and 1,068 [74.4%] females) fulfilling the criteria proposed by the International Headache Society for the diagnosis of migraine entered the study. Patients were treated with 10 mg of single-dose flunarizine (Sibelium) alone at bedtime in open fashion for 6 months. At the end of this treatment period, flunarizine was withdrawn, but the follow-up of the patients continued for another 6 months. The evaluation criteria used were a rating scale (the GES) based on frequency, duration, intensity, and characteristics of the attacks as well as a checklist of possible side-effects. This clinical assessment was recorded in detail in the patients' rating notebooks at the start and at the end of the third, sixth, ninth, and twelfth month of the study. A mean decrease of 66.9% in the GES was obtained at the end of the treatment period, which implies a good or excellent result in the prophylaxis of migraine attacks in 69.5% of the patients. This improvement was practically unchanged at the end of the follow-up period. Side-effects were moderate, the most frequent ones being drowsiness and weight gain. Their incidence decreased after the first months of treatment. PMID- 3052851 TI - Development and application of transseptal left heart catheterization. AB - The recent interest in transseptal left heart catheterization affords an opportunity to review the development and application of this procedure. This review briefly follows the history of right and left heart catheterization to the development of the transseptal procedure. A detailed description of the technique of transseptal catheterization is provided, as well as the indications and contraindications to its use. A learning curve is associated with this technique and is inversely related to the complication rate of the transseptal approach. Physicians working in high volume catheterization laboratories may gain the experience to safely perform transseptal left heart catheterization on carefully selected patients. PMID- 3052852 TI - A consensus sequence of three DNA replication terminus sites on the E. coli chromosome is highly homologous to the terR sites of the R6K plasmid. AB - Using the "Ter assay" we developed, three separate replication terminus (terC1, terC2, and terC3) sites on the E. coli chromosome were identified. The locations are at 28.3, 35.6, and 33.9 min on the linkage map, respectively. The terC1 site can block the counter-clockwise replication fork only, while the terC2 and terC3 sites inhibit the clockwise fork traveling on the chromosome. DNA sequences of the terC sites required for termination of DNA replication are highly homologous to those of terminus (terR) sites of the R6K plasmid, and the 21 bp consensus DNA sequence of terC is 5'-(A or T) TTAGTTACAACAT (A or C) CT (A or T) (A or T) (A or T) T-3'. In addition, all Ter active pUC-terC plasmids had a low copy number and were unstable in the host cells. PMID- 3052853 TI - A protein component of Drosophila polar granules is encoded by vasa and has extensive sequence similarity to ATP-dependent helicases. AB - Determinants of pole cells, which are precursors of the germ line, are provided maternally and are localized to the posterior pole of the Drosophila egg, as are polar granules. It has been hypothesized that certain RNA molecules associated with polar granules may be necessary for pole cell determination. Using a monoclonal antibody (Mab46F11) against polar granules, we have cloned the gene for one of their components. This gene turns out to be vasa, which is required maternally for the formation of polar granules and germ cells. This polar granule component shows significant sequence similarity to eIF-4A, a translation initiation factor that binds to mRNA, and to other helicases. PMID- 3052854 TI - A replication fork barrier at the 3' end of yeast ribosomal RNA genes. AB - Replication of the approximately 200 tandem copies of yeast ribosomal RNA genes (rDNA) is known to be initiated within a subset of the repeats, with transcription continuing during the replication process. To examine replication fork movement in this gene cluster, we used a two-dimensional (2D) agarose gel electrophoresis procedure that distinguishes molecules with different branched structures. Replication forks move through most of the rDNA in the same direction in which RNA polymerase I transcribes the 35S rRNA precursor: the 3' end of this transcription unit acts as a barrier to replication forks moving in the direction opposite to RNA polymerase I. The replication fork barrier (RFB) is observed as the accumulation of branched intermediates of specific size. We propose that the act of transcription may influence the movement of replication forks, creating barriers at the 3' ends of actively transcribed genes. PMID- 3052855 TI - Nuclear-organelle interactions: nuclear antisense gene inhibits ribulose bisphosphate carboxylase enzyme levels in transformed tobacco plants. AB - The biosynthesis of ribulose bisphosphate carboxylase (RUBISCO) provides a model system for studying the coordination of nuclear and organelle gene expression, since this abundantly transcribed and expressed chloroplast enzyme is composed of small (SS) and large subunits (LS) encoded by a nuclear multigene family and a single chloroplast gene, respectively. We have tested the possibility that SS mRNA or protein levels affect LS mRNA amounts or LS protein production and accumulation. We find that expression of antisense DNA sequences for the SS in transgenic tobacco plants drastically reduces the accumulation of SS mRNA and SS protein. These changes are accompanied by corresponding reductions of LS protein but not LS mRNA amounts; accumulation of the LS protein appears to be regulated by translational and posttranslational factors. We also find that the transgenic plants display striking variations in growth that are correlated with antisense gene dosage. PMID- 3052856 TI - Copper activates metallothionein gene transcription by altering the conformation of a specific DNA binding protein. AB - Copper homeostasis in yeast involves a copper binding protein, metallothionein, and a trans-acting regulatory protein that activates transcription of the metallothionein gene in response to copper ions. We show that the regulatory protein specifically binds to the metallothionein gene control sequences in the presence, but not in the absence, of copper. Both the DNA binding and metalloregulatory functions of the transacting factor are contained within its aminoterminal domain, and partial proteolysis experiments show that copper activates this domain by causing a major switch in its conformation. Silver also activates the DNA binding domain in vitro and induces metallothionein gene transcription in vivo. We propose a novel copper cluster model for the DNA binding domain based on its surprising structural similarities to metallothionein itself. PMID- 3052857 TI - Activation of B lymphocytes with human serum-treated zymosan. AB - C3 fragments fixed on zymosan particles were presented to resting human B lymphocytes. The opsonized zymosan (Ops-Z) particles induced release of leukocyte migration inhibitory factor, a slight decrease in mIgD, and a slight increase in the activation marker Blast-2. The B cells did not proceed further along the pathway of activation: they did not respond to B cell growth factor (BCGF) and Ops-Z did not synergize with other activators for BCGF response either. Thus, we found that interaction between C3 fragments and CR2 initiates the activation of human B lymphocytes, but this is limited to the early phase. PMID- 3052858 TI - Separation and characterization of human T lymphocytes with varying adhesiveness for endothelial cells. AB - Previous studies in this laboratory have demonstrated that the adhesion of T lymphocytes to endothelial cell (EC) monolayers in vitro can be increased by preincubation of the EC with interferon-gamma, interleukin 1 (IL-1), tumor necrosis factor-alpha (TNF), or lipopolysaccharide (LPS), or by stimulation of the T cells with phorbol esters. In this report, we have demonstrated that three subpopulations of human peripheral blood T cells can be identified on the basis of their abilities to bind to EC: (1) a strongly binding group which binds to unstimulated EC; (2) an intermediately binding subset which adheres to EC only if these cells have been stimulated with IL-1, TNF, or LPS; and (3) a weakly binding subpopulation which adheres poorly to either unstimulated or stimulated EC. The more adhesive subgroups had larger cellular volumes than the less adhesive cells, were relatively enriched in cells bearing the OKM1 surface marker, and expressed relatively greater amounts of the lymphocyte-function-associated-1 molecule. Stimulation of the EC to bind increased numbers of T cells by IL-1, TNF, and LPS appeared to be mediated by the expression of a common adhesion molecule on the EC. PMID- 3052859 TI - The birth of immunology: Metchnikoff, the embryologist. AB - Metchnikoff must be viewed first as an embryologist, who, influenced by the Darwinian currents of the 1860s and 1870s, sought to establish a genetic and embryologic unity in phylogeny. His principal early theory that the mesoderm was the origin of endodermal structures enabled him to extend the observation of mesodermal digestive processes to a theory of immunity. Observation of amoeboid phagocytosis was not novel, but Metchnikoff's scientific investigations had prepared him to interpret this activity as a manifestation of a generalized property of the mesoderm. Earlier observers noted the presence of microorganisms and particles in leukocytes, and the notion of phagocytosis had previously been entertained, but only Metchnikoff recognized the importance of phagocytosis in a general scheme of inflammation and to develop an experimental model for its investigation. The observation was thus viewed not solely as an issue of pathology, but rather as a contribution to Metchnikoff's general idea of genetic unity and his hypothesis of a primordial multicellular organism, Parenchymella, later called Phagocytella. It is striking that he ultimately viewed phagocytosis as a question of immunity, considering the context of his research activities, which had been confined to evolution and biology of development. To demonstrate how the famous Messina experiments were extended to a new theory of immunity requires formulating Metchnikoff's recognition of both the importance of phagocytosis for his mesodermal theory and a more general theory of pathology. The result was the genesis of a new idea, immunity. PMID- 3052860 TI - Single-visit root canal. PMID- 3052861 TI - Optical approaches to the dynamics of cellular motility. A symposium in honor of Robert Day Allen. October 5-8, 1987, Woods Hole, Massachusetts. Proceedings. PMID- 3052862 TI - The organization and regulation of the macrophage actin skeleton. AB - To move, leukocytes extend portions of their cortical cytoplasm as pseudopods. These pseudopods are filled with a three-dimensional actin filament skeleton, the reversible assembly of which in response to receptor stimulation is thought to play a major role in providing the mechanical force for these protrusive movements. The organization of this actin skeleton occurs at different levels within the cell, and a number of macrophage proteins have been isolated and shown to affect the architecture, assembly, stability, and length of actin filaments in vitro. The architecture of cytoplasmic actin is regulated by proteins that cross link filaments in higher-order structures. Actin-binding protein plays a major role in defining network structure by cross-linking actin filaments into orthogonal networks. Gelsolin may have a central role in regulating network structure. It binds to the sides of actin filaments and severs them, and binds the "barbed" filament end, thereby blocking monomer addition at this end. Gelsolin is activated to bind actin filaments by microM calcium. Dissociation of gelsolin bound on filament ends occurs in the presence of the polyphosphoinositides, PIP and PIP2. Calcium and PIP2 have been shown to be intracellular messengers of cell stimulation. PMID- 3052863 TI - Optical approaches to the study of foraminiferan motility. AB - Microtubules are the major cytoskeletal component of foraminiferan reticulopodia. Video-enhanced differential interference contrast light microscopy has demonstrated that the microtubules serve as the intracellular tracks along which rapid bidirectional organelle transport and cell surface motility occurs. Microtubules appear to move, both axially and laterally within the pseudopodial cytoplasm, and these microtubule translocations appear to drive the various reticulopodial movements. F-actin is localized to discrete filament plaques form at sites of pseudopod-substrate adhesion. Correlative immunofluorescence and electron microscopy reveals a structural interaction between microtubules and the actin-containing filament plaques. Our recent data on reticulopodial motility are discussed in an historical context, and a model for foram motility, based on motile microtubules, is presented. PMID- 3052864 TI - Progress in video microscopy. AB - The progress in video microscopy is reviewed from its early inception, especially with respect to improvements of the microscope image quality. Very recent advances that provide serial optical sections and depth of field as thin as 0.1 micron and that make possible the recording of birefringent images of individual microtubules (25 nm in diameter) directly in live, dividing cells are also documented. PMID- 3052865 TI - Biophysics of the leading lamella. PMID- 3052866 TI - Direct observation of mitotic spindle elongation in vitro. AB - Successful reactivation in vitro of anaphase B has recently been achieved with mitotic spindles isolated from the diatom Stephanopyxis turris. When a population of isolated spindles was studied indirectly by using immunofluorescence, nearly all of them were found to have elongated; however, when studied directly by using video microscopy, only a small proportion of spindles elongated. We report here conditions that allow nearly all of the spindles to elongate when observed directly with video microscopy. These direct observations validate previous ones made using indirect immunofluorescence. In addition, we find that the isolated spindles elongate with a linear rate, that the elongation is unchanged after the chromatin surrounding the spindles is digested with DNase I, and that during elongation a phase-dense matrix may accumulate in the spindle midzone. PMID- 3052867 TI - Detection of single fluorescent microtubules and methods for determining their dynamics in living cells. AB - The ability to tag biological molecules fluorescently and to detect their distribution in living cells has promoted the study of cytoplasmic organization in general and microtubule dynamics in particular. The techniques that we have selected and developed allowed the determination of spatial and temporal changes of the microtubule network in living fibroblasts at the level of individual microtubules. We have employed two general approaches for determining pattern changes: direct video microscopy and photobleaching and subsequent observation. Direct observation of fluorescent microtubules by high-definition video microscopy provided good spatial resolution at several time points, but was limited to the less congested and thinner periphery of the cell. This approach was made possible by a relatively bright, photostable reporter, xrhodamine tubulin, and showed that microtubules underwent rounds of assembly and disassembly from their ends. Bleaching and subsequent observation of lysed cells improved the signal to noise ratio by extracting soluble chromophore and permitted observations in congested areas, but was limited to a single time interval. This approach demonstrated that microtubule domains were replaced one by one and that turnover was most rapid at the cell periphery. Antibodies specific for nonbleached chromophore can be used to enhance the signal to noise ratio further or to extend spatial resolution by the use of immunoelectron microscopy. Direct video microscopy and photobleaching are two approaches to the study of dynamics that have complementary strengths and wide application to the biology of living cells. PMID- 3052868 TI - Isolation and reactivation of highly-coupled newt lung cilia. AB - The paired lungs of the newt, Taricha granulosa, are simple, unbranched sacs, 3.5 5.0 cm in length. The inner epithelium overlying the pulmonary vein is differentiated into a mucociliary tract that extends the entire length of the lung. Populations of single, demembranated ciliary axonemes, 12-13 micron in length, can be isolated by extracting whole lungs or primary cultures of the ciliated epithelium with Triton X-100. The motile capabilities of the isolated axonemes are the highest yet obtained for any ciliary model. When exposed to a suitable reactivating medium containing Mg2+ and ATP, nearly 100% of the axonemes become motile. Uniform reactivation of high quality requires short extraction times, minimization of mechanical damage, and strict adherence to optimal conditions throughout the extraction, storage, and reactivation procedures. Significant deviations from either pH 7.0 or 0.12 M salt can lead to a rapid, irreversible decrease in the beat frequency of reactivated axonemes. Both DTT and EDTA serve to stabilize their motility. The isolated axonemes beat at 29.5 Hz in the presence of 1.75 mM ATP at 21 degrees C, matching the beat frequencies measured for cultured cells at the same temperature. With 5 mM ATP, beat frequencies over 40 Hz are measured. Our results show that neither the plasma membrane, accessory structures, nor hydrodynamic coupling of cilia are required for this activity and imply that the lack of these factors is not responsible for the low motile capabilities of ciliary models isolated previously. PMID- 3052869 TI - Analysis of bacterial flagellar rotation. AB - Bacterial flagella have rotary motors at their base; embedded in the cytoplasmic membrane and powered by transmembrane ion gradients instead of ATP. Assays have been developed to measure the torque output of individual motors over a wide regime of load, to correlate the energizing proton flux with rotation speed and relate through genetic analysis motor structure to function. These assays promise substantial advances in understanding mechanochemical coupling in these motors. Here, I summarize the present status of our understanding of energy transduction in bacterial flagella and compare this with the case for muscle. PMID- 3052870 TI - Results obtained with a sensitive confocal scanning system designed for epifluorescence. AB - A wide variety of specimens has been examined with our apparatus, a commerical version of which is being manufactured by Bio-Rad/Lasersharp. The advantages expected of a confocal system have been realised in practice, the most striking advantage being the exclusion of glare from out-of-focus structures. This has made it possible to image cytological details in unflattened cells and intact tissues that were previously inaccessible to the light microscope. PMID- 3052871 TI - Actin polymerization and pseudopod extension during amoeboid chemotaxis. AB - Amoebae of the cellular slime mold Dictyostelium discoideum are an excellent model system for the study of amoeboid chemotaxis. These cells can be studied as a homogeneous population whose response to chemotactic stimulation is sufficiently synchronous to permit the correlation of the changes in cell shape and biochemical events during chemotaxis. Having demonstrated this synchrony of response, we show that actin polymerization occurs in two stages during stimulation with chemoattractants. The assembly of F-actin that peaks between 40 and 60 sec after the onset of stimulation is temporally correlated with the growth of new pseudopods. F-actin, which is assembled by 60 sec after stimulation begins, is localized in the new pseudopods that are extended at this time. Both stages of actin polymerization during chemotactic stimulation involve polymerization at the barbed ends of actin filaments based on the cytochalasin sensitivity of this response. We present a hypothesis in which actin polymerization is one of the major driving forces for pseudopod extension during chemotaxis. The predictions of this model, that localized regulation of actin nucleation activity and actin filament cross-linking must occur, are discussed in the context of current models for signal transduction and of recent information regarding the types of actin-binding proteins that are present in the cell cortex. PMID- 3052872 TI - High hydrostatic pressure effects in vivo: changes in cell morphology, microtubule assembly, and actin organization. AB - We present the first study of the changes in the assembly and organization of actin filaments and microtubules that occur in epithelial cells subjected to the hydrostatic pressures of the deep sea. Interphase BSC-1 epithelial cells were pressurized at physiological temperature and fixed while under pressure. Changes in cell morphology and cytoskeletal organization were followed over a range of pressures from 1 to 610 atm. At atmospheric pressure, cells were flat and well attached. Exposure of cells to pressures of 290 atm or greater caused cell rounding and retraction from the substrate. This response became more pronounced with increased pressure, but the degree of response varied within the cell population in the pressure range of 290-400 atm. Microtubule assembly was not noticeably affected by pressures up to 290 atm, but by 320 atm, few microtubules remained. Most actin stress fibers completely disappeared by 290 atm. High pressure did not simply induce the overall depolymerization of actin filaments for, concurrent with cell rounding, the number of visible microvilli present on the cell surface increased dramatically. These effects of high pressure were reversible. Cells re-established their typical morphology, microtubule arrays appeared normal, and stress fibers reformed after approximately 1 hour at atmospheric pressure. High pressure may disrupt the normal assembly of microtubules and actin filaments by affecting the cellular regulatory mechanisms that control cytological changes during the transition from interphase into mitosis. PMID- 3052873 TI - Cell-cycle modulation of MPM-2-specific spindle pole body phosphorylation in Aspergillus nidulans. AB - MPM-2 is a monoclonal antibody that interacts with mitosis-specific phosphorylated proteins in many different organisms. Immunocytochemistry of tissue culture cells has shown that MPM-2 stains centrosomes, chromosomes, kinetochores, and spindles. In this paper, we demonstrate that MPM-2 staining colocalizes with the spindle pole body (SPB) of Aspergillus nidulans and that SPB staining varies during the mitotic cycle. In an unsynchronized population, about one-fourth to one-third of the cells stain with MPM-2 at the spindle plaques or SPBs. Nuclei in mitosis have two SPBs localized at the ends of the spindle, both of which stain with MPM-2. To determine when MPM-2 staining appears, we have examined the effects of temperature-sensitive cell-cycle mutations that block nuclear division in S or G2. Only a very small fraction of cells blocked in S phase stain with MPM-2. In contrast, a large fraction of cells blocked in G2 stain brightly at the SPB. These data suggest that MPM-2 reactivity of SPBs appears in G2. Moreover, the fact that cells blocked in G2 showed MPM-2 staining but no spindles suggests that reactivity of SPBs occurs prior to mitosis but is not sufficient to trigger spindle formation. When G2-blocked cells were downshifted to permissive temperature, they generated a mitotic spindle with an SPB at each end. Both SPBs stained with MPM-2 in all of the mitotic cells. PMID- 3052874 TI - Colocalisation of acetylated microtubules, glial filaments, and mitochondria in astrocytes in vitro. AB - We have recently shown that acetylated alpha-tubulin containing microtubules (acetyl-MTs; labeled by antibody 6-11B-1) constitute a cold-stable subset of the microtubule network of nonneuronal cells in rat primary forebrain cultures [Cambray-Deakin and Burgoyne: Cell Motil. 8(3):284-291, 1987b]. In contrast, tyrosinated alpha-tubulin containing MTs (tyr-MTs; labeled by antibody YL1/2) are cold-labile. Here we have examined the distribution of acetyl-MTs and tyr-MTs in cultures of newborn rat forebrain astrocytes and simultaneously investigated the distribution of mitochondria and glial filaments. In double-label immunofluorescence experiments a marked colocalisation of acetyl-MTs and glial filament bundles was observed. Tyr-MTs did not show a similar colocalisation with glial filament bundles. Furthermore, the distribution of mitochondria closely followed that of the acetyl-MT and glial filament bundles. When cells were exposed to short-term (30-min) treatments with MT-disrupting agents such as colchicine and nocodazole, the tyr-MT network was removed but the distributions of acetyl-MTs, glial filaments, and mitochondria were unchanged. Increased exposure to colchicine (9-16 hr) caused a progressive disruption of the acetyl MTs and the collapse of glial filaments and mitochondria to the perinuclear region. These results suggest that acetyl-MTs and glial filaments but not tyr-MTs may be involved in the intracellular transport of organelles and/or in the control of their cytoplasmic distribution. PMID- 3052875 TI - Enzymatic barriers to peptide and protein absorption. AB - Continuing advances in biotechnology promise to provide a large number of peptides and proteins that would significantly expand the range of pharmaceuticals to treat diseases now poorly controlled. Even at this early stage, it is clear that the success of these entities as drugs of the future would depend, at least in part, on the success in overcoming the obstacles in their delivery. Chief among these are the ubiquitous enzymatic barriers. These include the site of administration, where the peptide is placed, the vascular endothelium that peptides must cross to enter the circulation, the blood that distributes the peptide to its target site, and the liver and kidneys where the peptide is metabolized and eliminated, respectively. The review examines: (1) the nature and efficiency of the enzymatic barriers in degrading peptides and proteins at various absorption sites, and (2) the strategies that can be used to perturb these barriers. These sites include the subcutaneous and intramuscular spaces and the intestinal, nasal, buccal, rectal, vaginal, and ocular surfaces. PMID- 3052876 TI - Alternative delivery systems for peptides and proteins as drugs. AB - The emergence of recombinant DNA technology has resulted in the large-scale production of a myriad of genetically engineered proteins and peptides, making many of them available for the first time for potential use as therapeutic entities. In addition, increased knowledge in the area of peptide/polypeptide hormones has resulted in an expansion of research efforts utilizing peptide synthetic chemistry, aimed toward developing novel therapeutic peptide drugs. Proteins and peptides cannot readily be administered by the conventional oral route, and thus alternative delivery methods to circumvent the necessity of frequent injections are being explored. This article reviews the current state-of the-art technology of such methodologies, encompassing localized administration, administration to various body cavities (i.e., nasal, rectal, etc.), as well as controlled-release injectable or implantable systems. These different approaches result in quite different pharmacokinetics that may in part dictate which approach is best suited for a particular protein or peptide. PMID- 3052877 TI - A proposed natural history of symptomatic anterior cruciate ligament injuries of the knee. AB - This article highlights the symptoms and clinical findings associated with anterior cruciate ligament instability. The symptomatic anterior cruciate ligament-deficient knee presents as an acute isolated incident or as a chronic progression of recurrent episodes of instability. The results of 71 patients diagnosed with an anterior cruciate ligament instability to the knee are reported. No distinction was made between the acute and chronic injuries as both patient populations received the same surgical procedure. The Lachman's test and the pivot shift test are the clinical examinations used to document cruciate ligament instability. The recommended procedure of choice for the symptomatic anterior cruciate-deficient knee is an autogenous patella tendon graft, bone tendon-bone preparation, arthroscopically placed in an isometric position within the knee joint. A rehabilitation program for the reconstructed knee is outlined. PMID- 3052878 TI - Anatomy and biomechanics of the anterior cruciate ligament. AB - The ACL is an important living soft-tissue component of ancient origin that acts in combination with other complex structures to provide control of femorotibial kinematics. The effect on the knee of its loss, resulting in disruptive kinematics and often subsequent degenerative changes, probably occurs not only because of its lack of structural integrity but also perhaps because of disruption of its proprioceptive function. The complexity of this ligament and associated normal kinematics of the knee challenges the ability of orthopaedists to devise effective therapeutic measures to reconstitute its function when lost. A clearer understanding of the normal role of the ACL will aid in this effort. PMID- 3052879 TI - Diagnosis of acute and chronic anterior cruciate ligament tears. AB - The diagnosis of acute and chronic injuries of the ACL has been reviewed. Attention has been given to the history, physical examination, and other diagnostic tools available for evaluation of the ligamentous injury. Clinical correlations have been highlighted. Finally, our recommendations for surgical intervention have been presented. PMID- 3052880 TI - Primary repair of the anterior cruciate ligament. AB - The article reviews the clinical features of the acutely torn anterior cruciate ligament. The alternatives and techniques of repair are discussed. Results of primary repair are presented. A rationale for therapeutic options is presented. PMID- 3052881 TI - The role of lateral extra-articular procedures for anterolateral rotatory instability. AB - The goal of any surgical procedure to correct the instability caused by loss of the ACL is to control the abnormal anterior excursion of the tibia on the femur. Because the main problem is loss of the ACL, it would seem most reasonable to approach this problem by performing an intra-articular reconstruction of the ACL, thus approximating as closely as possible the normal anatomy of the ACL. The classic open intra-articular ACL reconstructions are technically demanding surgical procedures that usually require a significant "learning curve" to achieve a level of technical expertise and confidence. In addition, postoperative complications such as adhesions, loss of motion, prolonged muscle atrophy, and a long rehabilitation period are well known. Thus, it would appear that the extra articular reconstructive procedures for the anterior cruciate-deficient knee would offer some advantage over these more formidable surgical procedures. Whereas the main problem is certainly the loss of the ACL, the extra-articular procedures are directed more toward the most symptomatic anterior excursion of the tibia on the femur, the pivot shift phenomenon, where the anterolateral portion of the tibia moves anterior in relation to the femur. Thus, the goal of the extra-articular reconstructive procedures for anterolateral rotatory instability is to eliminate functional instability. These goals are most readily achieved by positioning some portion of the iliotibial tract posterior to the transverse center of rotation of the knee to provide a reinforcement for the lateral tibial plateau as the knee approaches terminal extension. All of the extra-articular procedures discussed in this article have been used successfully as reported by the various authors. There are many technical details inherent in each of these surgical procedures, and the reader is referred to the original articles for a more explicit description of these surgical procedures. For the individual surgeon to participate in and view the actual surgical procedure that he or she intends to perform would be the ideal situation. Various workshops where surgical procedures of the knee are actually performed and studied are currently available and are of great value to the surgeon. Of equal importance to the technical demands of the various surgical procedures is selection of the appropriate procedure for each patient. The selection must be based on many factors. The most important factor is the identification of the patient with a high level of athletic activity who is unwilling to modify his or her activity level to compensate for a deficient ACL.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3052882 TI - Synthetic and allograft anterior cruciate ligament reconstruction. AB - Although still in the early stages of development, the use of synthetics and allografts in ACL surgery appears promising. Two prosthetic ligaments, the Kennedy LAD and Gore-Tex, are FDA approved for limited indications. The Kennedy LAD has been shown to be effective in augmenting an autograft with inherent structural weakness in its central portion. The proposed benefits of using this device with grafts of greater strength are unproved. The Gore-Tex ACL reconstruction allows a rapid initiation of vigorous rehabilitation and return to full activities. However, the complication rate with this procedure appears to be higher than that with autograft reconstruction. The use of allograft for ACL reconstruction also has many potential advantages and short-term clinical trials have shown good results. However, the benefits must be weighed against the possibility of long-term failure and potential spread of infectious disease. PMID- 3052883 TI - Trends in rehabilitation following anterior cruciate ligament reconstruction. AB - In summary, the use of an autogenous intra-articular reconstruction has been identified as an effective treatment modality to stabilize the ACL-deficient knee. However, despite the fact that rehabilitation programs are becoming more aggressive and less time consuming, the patient who elects to have his knee stabilized surgically continues to face a long recovery period. Continued research in surgical and rehabilitative management is necessary to reduce this time and expense if ACL reconstruction is to be practical for all. PMID- 3052884 TI - Principles of bracing for the anterior cruciate ligament-deficient knee. AB - The preceding discussion has profiled the three different types of knee braces available on today's market. It has attempted to discuss the controversies surrounding these braces and to analyze the scientific data presented to date. Prophylactic braces have been shown to be ineffective in preventing knee injuries in their present-day design. Evidence has also shown that their use may even lead to increased knee injuries. On the other hand, rehabilitative braces do serve a useful purpose in regard to the operative and nonoperative treatment of ligamentous knee injuries. With their use in the application and control of joint motion, they are an important addition to the surgeon's armamentarium. One must keep in mind, however, that these braces provide little static anterior/posterior control and the hinge settings may not actually reflect true joint motion. Functional knee braces may play a role in the treatment of patients with pathologic laxity owing to an injury of the ACL. Combined with an adequate rehabilitation program and activity modification, these braces do limit excessive anterior tibial translation under low-loading conditions. However, under conditions of high loading these braces provide little or no resistance to anterior translation. Therefore, in most sporting activities, their efficacy is questionable. Knee bracing continues to be a complex and controversial topic in the field of orthopaedic surgery. The answers for the design of the "ideal" brace are being continually worked out and the need for more detailed, well-controlled studies continues to be great.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3052885 TI - Complications of intra-articular anterior cruciate reconstruction. AB - Intra-articular reconstruction of the ACL is a powerful technique, but is associated with a variety of potential complications. Careful patient selection, precise intraoperative technique, and aggressive rehabilitation can help minimize these problems. Our most common complication, postoperative limitation of motion, was nearly eliminated by a change to arthroscopic surgical technique and early motion. PMID- 3052887 TI - [Mitochondrial transport of cations]. PMID- 3052886 TI - [Tryptophan metabolism by the indoleacrylic acid pathway]. PMID- 3052888 TI - [An improved guide for electrodes for stereotaxic brain surgery]. PMID- 3052889 TI - [History of pediatric neurology in Czechoslovakia]. PMID- 3052890 TI - [Bloch's multidimensional drawing test in clinical practice]. PMID- 3052891 TI - [Professor Karel Kuffner and his textbook of psychiatry]. PMID- 3052892 TI - [The building and rebuilding of the state hospital in Bratislava in the first decade of its activities as a teaching center at Comenius University Medical School]. PMID- 3052893 TI - Prophylaxis in gynaecological and obstetric surgery: a comparative randomised multicentre study of single-dose cefotetan versus two doses of cefazolin. AB - Antimicrobial prophylaxis is recommended in all clean-contaminated surgery where the critical threshold of number and virulence of the contaminating organisms with respect to host resistance is reached. Obstetric and gynaecological surgery is clean-contaminated and risk of infection due to aerobic and anaerobic bacteria without prophylaxis can be quantified at 30-40% for vaginal hysterectomy, 10-35% for abdominal hysterectomy and 10-34% for caesarean section. To assess the role of two different cephalosporins as short term prophylaxis, we carried out a multicentre randomised study involving a single 2 g i.v. dose of cefotetan in comparison with two doses of cefazolin (2 g i.v. before surgery and after 8 hours). Criteria for exclusion were: exposure to antibiotics within 7 days, preoperative infection, hypersensitivity to beta-lactams. Four hundred and sixty patients entered the study, of which 229 received cefotetan and 231 cefazolin. No significant differences in mean age, obesity, preoperative weight loss, diabetes, type of disease, type of surgery (vaginal or abdominal hysterectomies and caesarean sections) and number of pregnancies and abortions existed between the two groups of patients. The total rate of infected patients undergoing hysterectomy was 8.6% (13/151) in the cefotetan group and 17.4% (29/167) in the cefazolin group (p less than 0.05). This difference was due to cases of symptomatic bacteriuria and antibiotic retreatment, while wound infections were not significantly different (2.6% and 1.8% respectively). Among patients undergoing caesarean section, 9 of 78 (11.5%) and 7 of 64 (10.9%) were infected following cefotetan and cefazolin, respectively (not significant). Cefotetan mean tissue concentrations in gynaecological organs were higher than those of cefazolin (25.5-44.8 vs. 7.4-9.5 mg/kg).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3052894 TI - Comparative clinical and pharmacokinetic aspects of cefotetan versus cefoxitin plus metronidazole in vaginal hysterectomy. AB - In a randomised clinical trial, 102 women who underwent vaginal hysterectomy were given a single preoperative 2g dose of cefotetan (CTT) (52 pts) or three perioperative 2g doses of cefoxitin (CFX) plus 0.5g metronidazole (50 pts) as antibiotic prophylaxis. No statistically significant differences between the groups were detected in the clinical response (100% for both groups). The incidence of major wound infection (2% CTT and 0% CFX) were also comparable between the two treatment groups; post-operative changes in body temperature, duration of hospitalisation and post-operative urinary tract infections (16% CTT and 20% CFX) were similar. Both drugs were well tolerated. Twenty nine of the 102 patients were further investigated to determine the pharmacokinetics following a single 2g dose of CTT or CFX, in both serum and tissue. Although both antibiotics provided good concentrations during the early phase of surgery, CTT levels persisted for a longer time period. These results confirm that single dose cefotetan is equally as effective and safe as multiple dose cefoxitin plus metronidazole for prophylaxis in patients undergoing vaginal hysterectomy. PMID- 3052895 TI - [Bacteriologic characteristics of 25 strains of Vibrio cholerae, biotype eltor isolated in Mali]. AB - This work studies bacteriological characters of 25 strains of V. cholerae isolated in Mali during the outbreak of 1984-1987. These strains were precisely identified by biochemistry characters, toxin secretion, presence of a structure considered as attachment factor. Antibiotic susceptibility study confirmed the great sensibility of V. cholerae to this products, and allowed to classify them according to their Cl90. This contribution of the laboratory is very interesting for the control of the disease and the choice of standardized scheme of treatment. PMID- 3052896 TI - [2 hemocultures positive for Vibrio cholerae]. AB - The authors report two cases of Vibrio cholerae isolation in hemocultures even though the copro-cultures where negative. In both cases, post-operated patients are concerned. The first one had supported a renal transplantation and was under immuno-depressor. The second one, after an aorto-aneurysm surgical treatment suffered from a mesenteric infarct. During the disease the vibrio was found in the hemoculture. PMID- 3052897 TI - [Low levels of chloroquine resistance of Plasmodium falciparum in the province of Zou in Benin]. AB - An in vivo test using the WHO protocol (chloroquine 25 mg/kg within 3 days, trial over 7 days) was performed in 72 children in the province of Zou, Benin, in July August 1987. The blood concentration of chloroquine was dosed before, during and after treatment by a sensitive method. This study showed a low rate of drug resistance (4.2%), even though surveys in Cotonou exhibited a high level of chloroquine resistance. PMID- 3052898 TI - [In vitro sensitivity of Plasmodium falciparum to chloroquine in the highland region of Madagascar in 1987]. AB - With the Le Bras's isotopic semi-microtest method, the authors have studied 139 strains of Plasmodium falciparum isolated in a village near Tananarive in the Highlands of Madagascar. Conditional malariometric rates show the increase of the recrudescence of Malaria in the area which was recently considered as "surveillance area". 5.8% of the 139 studied strains show an in vitro resistance higher than 120 nmoles/l, but whose the level is ever lower than in Africa. 4.3% exhibit a level between 90 and 120 nmoles/l. These values give better prospect for the Isle because the resistance strain rates have remain nearly stable since 1982. The resistance level is always low. PMID- 3052899 TI - [The severity of airport malaria]. AB - Report of a new case of airport malaria with renal failure. The evolution of the thirty cases previously described is reviewed. Most of the time, airport malaria seems to be a severe infection. PMID- 3052900 TI - [Epidemiologic study of malaria in the foothill area of the Taez region of the Arabic Republic of Yemen]. AB - Longitudinal Survey on Malaria is conducted in the Foothills (Ouadi Rissian) in an area between 450 m et 750 m elevation south-west of Taez in Yemen Arab Republic (Y.A.R.) Malaria is mesoendemic and transmission is perennial. P. falciparum is the only parasite species identified and A. arabiensis the main vector collected. Control measures are discussed. PMID- 3052901 TI - [Hydatid cyst of the thyroid. Apropos of a case in the Republic of Niger]. AB - Thyroid echinococcosis has been rarely found, especially in areas where hydatic cysts are rarely encountered. The authors report on a case of thyroid echinococcosis in a patient living in sahelian Africa (Niger), which is classically known as a region of low endemia for hydatic disease. PMID- 3052902 TI - [Stereotaxic brain biopsy as basis of therapy planning]. PMID- 3052903 TI - The impact of toxicity on carcinogenicity studies: implications for risk assessment. AB - This paper explores the inter-relationship between toxicity, genotoxicity, and carcinogenicity in laboratory rodents. To our knowledge this is the first attempt to integrate these factors and evaluate their implications for the process of risk assessment. The evaluation is based on information obtained from 2-year laboratory-animal studies involving 99 chemicals. The data suggest that only seven of the 53 positive carcinogenicity studies exhibited the types of target organ toxicity that could have been the cause of all observed carcinogenic effects. Furthermore, no apparent difference in mutagenicity as measured by the Ames Salmonella assay was observed between 'high dose only' carcinogens and the entire set of carcinogens. These findings suggest that the number of chemical carcinogens that we can identify solely through rodent studies as being potential tumor inducers through some indirect mechanism is small. Generally speaking, the identification of histopathological effects is not sufficient in itself for justifying mechanistic assumptions, and supplemental biological information will be necessary to reach definitive conclusions. PMID- 3052905 TI - [Comparative experimental studies on Plasmodium vivax isolated from south Yunnan and northwest Hunan]. PMID- 3052904 TI - Intracoronary infusion of prostaglandin I2 attenuates arterial baroreflex control of heart rate in conscious dogs. AB - Prostaglandin I2 (PGI2) is known to stimulate ventricular C fiber receptors resulting in a Bezold-Jarisch-like reflex. Also, cardiac receptor stimulation is known to interact with the expression of arterial baroreflexes. Therefore, experiments were performed to determine the effects of left circumflex coronary artery infusion of PGI2 on the baroreflex control of heart rate in conscious instrumented dogs. Dogs were instrumented chronically with an aortic catheter for the measurement of mean aortic pressure, hydraulic occluder cuffs on the descending aorta and inferior vena cava, a left ventricular catheter for the measurement of left ventricular pressure and heart rate, and a nonocclusive catheter in the left circumflex coronary artery. At the time of experimentation, arterial pressure was altered randomly in steps by partially inflating the occluders. Mean arterial pressure-heart curves (baroreflex curves) were constructed by fitting the data to a logistic curve by nonlinear regression. PGI2 infused into the left circumflex coronary artery at doses of 10, 20, and 50 ng/kg/min caused significant (p less than 0.05) inhibition of the maximum heart rate, heart rate range, and maximum slope of the curve compared to the control baroreflex curve obtained during intracoronary infusion of PGI2 vehicle. PGI2 had no significant effect on the minimum heart rate during hypertension. Since PGI2 is known to stimulate left ventricular receptors, these effects were most likely produced via stimulation of cardiac receptors. In additional experiments using beta 1-blockade with metoprolol or cholinergic blockade with atropine methyl bromide, it was shown that PGI2 attenuates baroreflex-mediated tachycardia by preventing parasympathetic withdrawal completely and by attenuating sympathetic stimulation by approximately 50%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3052907 TI - [Scanning electron microscopic observation on the effects of pyquiton and albendazole on the tegument of Clonorchis sinensis]. PMID- 3052906 TI - [Plasmodium falciparum: correlation between immunofluorescent properties of monoclonal antibodies and their protective activities]. PMID- 3052908 TI - [Observation on the inhibitory effect of ketotifen, cyproheptadine and pizotifenum on Plasmodium falciparum in vitro]. PMID- 3052909 TI - [Dot-immunobinding assay with sheep hydatid fluid antigen for the diagnosis of hydatidosis]. PMID- 3052911 TI - Restorative materials. PMID- 3052910 TI - [Study on the role of low density microfilaremia cases in the transmission of filariasis in late stage of filariasis control]. PMID- 3052912 TI - Current concepts of mitral valve reconstruction for mitral insufficiency. AB - In recent years, there has been a renewed interest in surgical reconstruction of the insufficient mitral valve because of reconfirmation of the limitations of existing prosthetic and bioprosthetic valves. A follow-up study, including late functional data, of 148 patients who underwent mitral valve reconstruction at our institution was combined with a review of the literature to assess the current status of mitral reconstruction. The results indicate that mitral reconstruction by Carpentier techniques is widely applicable, durable, and relatively free of complication. Freedom from late thromboembolic and anticoagulant complications is particularly notable. These factors could prove to justify earlier operative intervention in patients with mitral insufficiency before permanent myocardial damage evolves. As mitral valve reconstruction techniques become more familiar and widely used, mitral reconstruction may become the operative procedure of choice for mitral insufficiency, especially insufficiency due to degenerative disease. PMID- 3052913 TI - Current therapy of the failing heart. AB - Myocardial dysfunction eventuating in systolic and diastolic pump function abnormalities is a consequence of a wide variety of cardiac diseases. The symptoms that develop in this syndrome appear to be related as much to peripheral and neurohormonal mechanisms as to the underlying pathological and cardiac functional abnormality. Relief of symptoms, slowing of the progression of the cardiac functional abnormality, and prolongation of life provide the major agenda for the physician faced with the management of these patients. Judicious use of vasodilators, diuretics, digoxin, dietary therapy, and exercise therapy can relieve symptoms and improve the quality of life in most patients suffering from this syndrome. Recent evidence that vasodilator drugs can prolong life now provides the physician with further justification for routine use of this class of compounds. The eventual solution to the high mortality in this common disease process may be prevention of the development of overt heart failure by more prompt recognition and early treatment of the signs of ventricular dysfunction. This possibility must await the completion of current and proposed clinical trials. PMID- 3052914 TI - Quantifying valvular regurgitation. Limitations and inherent assumptions of Doppler techniques. PMID- 3052915 TI - Left ventricular function after correction of chronic aortic regurgitation. PMID- 3052916 TI - Intracavitary right heart cooling during coronary bypass surgery. A prospective randomized trial. AB - Augmented right heart cooling (RHC) with bicaval cannulation, pulmonary artery venting, and intracavitary cooling has been advocated for prevention of right ventricular failure and supraventricular tachyarrhythmias after open heart surgery. To evaluate RHC, 78 patients undergoing coronary bypass surgery were prospectively randomized to receive added RHC (n = 38) or standard protection with single atrial cannulation (SC) (n = 40). RHC and SC patients were similar regarding right coronary artery occlusion (n = 10 and 12, respectively), number of grafts performed (3.7 +/- 1.0 and 3.4 +/- 0.9), and cross-clamp time per graft (10.2 +/- 1.8 and 9.8 +/- 2.3 minutes). RHC led to significantly lower right atrial (11.6 degrees +/- 1.0 degree vs. 19.5 degrees +/- 3.3 degrees C) and right ventricular (7.2 degrees +/- 1.9 degrees vs. 12.2 degrees +/- 1.9 degrees C) temperatures. There was no detectable deterioration in right heart function or left heart function in either group after cardiopulmonary bypass. Bypass time was longer in RHC patients (86.7 +/- 17.9 vs. 76.0 +/- 18.2 minutes, p less than 0.05). Technical problems related to multiple cannulation occurred in four RHC patients. After cross-clamp removal, creatine kinase-MB levels were significantly higher with RHC at 2 hours (14.2 +/- 7.6 vs. 6.4 +/- 4.6 IU/l, p less than 0.001), 12 hours (19.1 +/- 19.5 vs. 8.6 +/- 10.3 IU/l, p less than 0.005), and 24 hours (14.1 +/- 19.6 vs. 7.1 +/- 9.2 IU/l, p less than 0.05). Mortality and morbidity were similar in the two groups. In particular, supraventricular tachyarrythmias occurred in 11 (28.9%) RHC and 10 (25%) SC patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3052917 TI - Experimental and clinical study of crystalloid cardioplegic solution in neonatal period and early infancy. Effects of calcium and prostacyclin analogue. AB - The effects of calcium and a prostacyclin (PGI2) analogue in the glucose-insulin potassium (GIK) cardioplegic solution for the neonatal period and early infancy were evaluated. The assessment was based mainly on semiquantitative scoring of mitochondrial damage and intracellular edema in postreperfusion biopsies. Experimentally, 45 isolated perfused newborn rabbit hearts (age, 0-2 days) underwent 2 hours of global ischemia at 15 degrees C with a single dose of GIK cardioplegic solution and were subsequently assigned to three groups: Group 1 hearts (n = 15) were infused with basic GIK cardioplegic solution alone (no added calcium, but measured at 0.1-0.2 mM/l); Group 2 hearts (n = 15) received GIK cardioplegic solution with calcium (1.2 mM); and Group 3 hearts (n = 15) received GIK cardioplegic solution with calcium and a PGI2 analogue (OP-41483, 300 micrograms/l). Group 3 hearts showed significantly lower mitochondrial damage and intracellular edema scores than did Group 1 and Group 2 hearts (p less than 0.05). Hemodynamic measurement (aortic flow and coronary flow) results after reperfusion were also better in Group 3 hearts than in the hearts of the other two groups (p less than 0.05). In the clinical study with 18 infants who were less than 3 months old, the same three cardioplegic solutions were used. Group 2 (n = 6) and Group 3 (n = 5) infants showed significantly lower mitochondrial damage scores than did Group 1 (n = 7) infants. Group 3 infants also showed significantly lower intracellular edema scores than did Group 1 and Group 2 infants.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3052918 TI - Reversal of recalcitrant cardiac allograft rejection with methotrexate. AB - Refractory cardiac transplant rejection is a major therapeutic dilemma. The effectiveness of methotrexate (MTX) in autoimmune diseases prompted us to explore its efficacy in 10 cardiac transplant recipients, aged 20-53 years (39 +/- 13 years; mean +/- SD), with biopsy evidence of drug-refractory cardiac allograft rejection. Nine cardiac transplant recipients were maintained on triple antirejection therapy (cyclosporine, azathioprine, and prednisone), and the remaining recipient was maintained on cyclosporine and prednisone. Rejection episodes treated with MTX occurred 20-422 days (165 +/- 137 days) after transplantation and were the sixth episode of rejection for one recipient, the third for four recipients, the second for four recipients, and the first for one recipient. Before MTX administration, cardiac allograft rejection persisted despite intensified immunosuppression including OKT3 antibody. MTX, given intravenously, orally, or by both routes at a dose of 10-175 mg (85 +/- 62 mg), reversed rejection in nine of 10 recipients (90%) within 7-63 days (26 +/- 18 days). Elevated pulmonary artery wedge pressures were reduced to normal levels after MTX therapy (15.2 +/- 3.5 before vs. 10.6 +/- 3.0 mm Hg after; p less than 0.05). Leukopenia occurred in five cardiac transplant recipients after treatment with MTX. Adverse reactions to MTX resolved after MTX therapy was discontinued in all but one recipient. This recipient received one of the larger MTX doses (150 mg) and developed fatal Pseudomonas pneumonia. Twelve moderate rejection episodes recurred in nine recipients, seven episodes of which were successfully re-treated with MTX. Two of these seven recipients have now been rejection-free for 15 months. Of five recurrent episodes of cardiac transplant rejection not treated with MTX, two could not be reversed and were fatal. MTX may be a valuable drug for reversing refractory cardiac allograft rejection. Before MTX therapy gains wider use, however, a clearer understanding of its enhanced ability to suppress the bone marrow is needed. PMID- 3052919 TI - Staged cardiac transplantation. Total artificial heart or ventricular-assist pump? AB - Because of donor organ unavailability, staged cardiac transplantation has been performed in eight patients with The Pennsylvania State University pneumatic total artificial heart (two patients) or with the Pierce-Donachy ventricular assist pump (six patients). Of the six patients who received the ventricular assist pump, four received cardiac allografts after 3, 11, 21, and 31 days of pump support, respectively. Two patients died before transplantation; the causes of death were non-device-related complications. One additional patient died of Pseudomonas sepsis after staged cardiac transplantation. The remaining three patients are alive and have been followed up for as long as 2 years after staged cardiac transplantation. Of the two patients who were supported with the total artificial heart, one underwent staged cardiac transplantation after 11 days of support. Unfortunately, this patient succumbed to fungal sepsis 17 days later. The remaining patient, who received the total artificial heart after rejection of a transplanted heart, expired 379 days later before a suitable donor organ could be located. These experiences indicate that 1) the ventricular-assist pump and total artificial heart can provide reasonably safe effective circulatory support until a patient's overall physiological status is optimal, until a donor organ can be located for transplantation, or both; 2) there is a need for short-, intermediate-, and long-term support system capabilities; and 3) regardless of the patients' underlying pathology (ischemic versus nonischemic cardiomyopathy), in most instances, the simpler external ventricular-assist pump is capable of satisfactory hemodynamic circulatory assistance. PMID- 3052920 TI - Minnesota ALG. Safe and effective immunosuppression for cardiac transplantation. AB - Fifty-six patients undergoing orthotopic cardiac transplantation were given Minnesota ALG prophylactically or therapeutically for acute cardiac rejection. During a follow-up period of 0-28 months (mean follow-up period, 11.9 months), the actuarial survival for the entire group was 96% and 86% at 30 days and 1 year, respectively. Actuarial freedom from rejection was 60% and 28% at 30 days and 1 year, respectively. All but seven rejection episodes responded to initial steroid pulses or a modification of a maintenance cyclosporine and azathioprine regimen. The seven failures were rescued with further Minnesota ALG therapy. Few serious hematologic or allergic reactions to Minnesota ALG were observed, and no new malignancies occurred during the follow-up period. We conclude that Minnesota ALG is safe and effective in cardiac transplantation. PMID- 3052922 TI - Magnetic resonance imaging with gadolinium-DTPA for detecting cardiac transplant rejection in rats. AB - To date, no noninvasive tool has gained widespread acceptance as an adequate substitute for endomyocardial biopsy for the diagnosis and grading of cardiac transplant rejection. We examined the potential role of magnetic resonance imaging with gadolinium (Gd)-diethylenetriamine penta-acetic acid (DTPA) image enhancement for the diagnosis of cardiac graft rejection. We studied 15 rats with heterotopic cardiac transplants, nine of which received no immunosuppression, and six of which received cyclosporine, azathioprine, and methylprednisolone. The animals underwent magnetic resonance imaging, which was immediately followed by sacrifice (2-12 days after transplant). Myocardial image enhancement was assessed on T1-weighted images performed before and after administration of Gd-DTPA, 0.5 mmol/kg. Histological specimens were graded I, II, or III to indicate increasing severity of rejection. In the absence of rejection, Gd-DTPA induced mild homogeneous myocardial enhancement. Ten of 11 cases with Grade II or III rejection manifested one or more areas of intense myocardial enhancement. The extent and distribution of intense myocardial enhancement corresponded to the severity and distribution of histological rejection. Quantitative myocardial enhancement, expressed as the ratio of maximal signal intensity after Gd-DTPA to signal intensity before Gd-DTPA administration, separated Grade I animals (1.61 +/- 0.27; mean +/- SD) from Grades II (2.89 +/- 0.58) and III (3.10 +/- 0.77; p less than 0.01) animals. In conclusion, cardiac transplant rejection is characterized by intense T1-weighted image enhancement after administration of Gd DTPA. Magnetic resonance imaging with Gd-DTPA thus has potential application in the clinical diagnosis of cardiac transplant rejection. PMID- 3052921 TI - Effect of preoperative hemodynamic support on survival after cardiac transplantation. AB - The accessibility and success of cardiac transplantation promotes transplantation for a broad range of recipients, including those requiring intravenous inotropes or mechanical-assist devices. To determine if survival is dependent on preoperative requirements for hemodynamic support, we studied 230 patients who underwent transplant at the Loyola, Stanford, and UTAH programs from December 1, 1984 through November 30, 1986, and who were followed up for 34 months postoperatively. Group 1 (n = 132 of 230, 57%) patients required only oral medical therapy to maintain hemodynamic compensation; Group 2 (n = 69 of 230, 30%) patients were dependent on intravenous inotropic support; and Group 3 (n = 29 of 230, 13%) patients required mechanical assistance. Pretransplant characteristics showed that dilated cardiomyopathy was more common in Group 2 patients, and lower cardiac index and ejection fraction were more prevalent in Group 3 patients as expected. Although survival was lower in Group 3 only at 1 month (Group 1, 98.5%; Group 2, 92.8%; and Group 3, 86.2%; p less than 0.01), the survival advantage in Groups 1 and 2 was lost by 3 months, with 1-year survival rates of 88.6% in Group 1, 81.2% in Group 2, and 82.8% in Group 3. Allograft survival and cause of death were not different among the three groups. Acute rejection occurred at a lower monthly frequency in the first 4 months in Group 3 (Group 1, 0.47 +/- 0.03; Group 2, 0.47 +/- 0.05; and Group 3, 0.29 +/- 0.06; p less than 0.01), whereas infectious complications occurred at similar frequencies.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3052923 TI - Echocardiography-guided endomyocardial biopsy. A 5-year experience. AB - The number of cardiac transplant procedures performed each year continues to increase and may exceed 1,600 procedures in 1987. In these patients, the diagnosis of rejection is obtained from serial endomyocardial biopsy. The absence of clinical symptoms associated with allograft rejection with cyclosporine therapy has necessitated a predetermined routine biopsy frequency that averages 10-15 biopsies per patient in the first posttransplant year. Traditionally, fluoroscopy has been used to guide the biopsy, but this technique has a number of negative features, including cumulative radiation exposure in the physician and the patient, limited portability, and the limited area of access (intraventricular septum) for biopsy. In contrast, echocardiography provides greater portability and flexibility for location of procedure performance, eliminates radiation exposure, provides important information about cardiac function, and safely allows biopsy of any area of the right ventricle, including the free wall and apex. We performed over 4,700 individual biopsies for evaluation of rejection in 58 patients who underwent orthotopic cardiac transplantation. Fluoroscopy was not required for any of the echocardiography guided biopsies, and only two complications occurred in 4,700 biopsies. We propose, therefore, that echocardiography is a useful alternative or adjunct to fluoroscopy for guiding endocardial biopsy and is a technique that can be learned easily by anyone now performing biopsies. PMID- 3052924 TI - Enzyme immunosorbent assay of prolactin with penicillinase as label. AB - This competitive, rapid, and sensitive enzyme immunosorbent assay for measuring human prolactin in plasma involves penicillinase (EC 3.5.2.6) as label. Microtiter plate wells coated with goat anti-rabbit gamma globulin are filled with antibody to prolactin and plasma sample or reference prolactin and incubated with penicillinase-labeled prolactin for 1 h at 37 degrees C. The enzyme activity of the bound complex is then measured. The assay is as sensitive as radioimmunoassay, the detection limit being 2.5 ng of prolactin per milliliter of plasma. The plasma prolactin values obtained by enzyme immunoassay correlated well with those determined by radioimmunoassay: r = 0.98, slope = 1.00, intercept = 1.11 ng/mL (n = 53). PMID- 3052925 TI - Immunoassay of alpha 1-acid glycoprotein in the Cobas Bio centrifugal analyzer. AB - There is an increasing demand for quantification of serum alpha 1-acid glycoprotein (AAG, orosomucoid) in studies evaluating the protein binding of highly bound basic drugs. This paper describes an adaptation of an automated immunoturbidimetric assay for this protein to the Cobas Bio centrifugal analyzer. Replicate analyses of aliquots from six different solutions were used in determining precision. We also analyzed 367 patients' serum samples, in duplicate, to determine the distribution of AAG in hospitalized patients. The intra- and inter-run CVs ranged from 1.3% to 4.4% and from 0.6% to 6.6%, respectively. AAG concentrations in patients' samples ranged from 0.38 to 3.16 g/L. Results by this method correlate well with those by radial immunodiffusion, with no significant amount of bias between the two methods. This immunoturbidimetric procedure is faster and less expensive than currently used radial immunodiffusion techniques, and precision is acceptable. PMID- 3052926 TI - Homogeneous time-resolved fluoroimmunoassay of thyroxin in serum. AB - We describe a rapid, simple nonseparation fluoroimmunoassay for determination of thyroxin in serum. The assay is based on the labeling of thyroxin directly with a fluorescent europium chelate, the fluorescence of which is quenched on binding to an antithyroxin antibody. With the assay buffer we used, maximum quenching is 90%. The rapid achievement of equilibrium in the assay solution, regardless of the sequence of reagent additions, allows fast measurement of thyroxin. Precision was good (CV less than 5%) within the clinical range for total thyroxin (50-300 nmol/L), and results correlated well with those by a commercial radioimmunoassay. PMID- 3052927 TI - Determination of D-sorbitol in human erythrocytes by an improved enzymatic method with fluorometric detection. AB - In this enzymatic method to analyze erythrocytes for D-sorbitol, the erythrocytes were separated from plasma by centrifugation, washed with isotonic saline (9 g/L NaCl), then diluted threefold with more saline. We lysed 1.5 mL of the diluted erythrocytes with chloroform and precipitated the protein with 58 g/L HClO4 solution. The resulting supernates were mixed with buffer, NAD+, and sorbitol dehydrogenase (EC 1.1.1.14). For standards, we added sorbitol to the erythrocytes before lysing. After 25 min of incubation at 37 degrees C, the fluorescence responses were recorded. Results for sorbitol were corrected for sample-blank and enzyme fluorescence, and were then normalized for hemoglobin content. Response was linear for a sorbitol concentration range of 1 to 30 mumol/L. Reproducibility was good, with an average CV of 2.5% at 10 mumol/L. Results for healthy individuals and diabetic patients are presented. PMID- 3052928 TI - Adulterants causing false negatives in illicit drug testing. AB - Illicit-drug users may attempt to falsify results by in vitro adulteration of specimens. We investigated eight additives (NaCl, Visine, handsoap, Drano, bleach, vinegar, golden-seal tea, and lemon juice) claimed by drug users to invalidate enzyme immunoassay (EIA) drug assays. We also analyzed adulterated urine specimens to determine if they could be identified, adding adulterants at several concentrations to 222 EIA-positive specimens confirmed by gas chromatography and mass spectrometry (GC/MS) to contain illicit drugs. To identify adulterated urines, we monitored pH, relative density, and urine color and turbidity at adulterant concentrations that falsified EIA results. Specimens contaminated with NaCl had relative densities greater than 1.035. Liquid Drano, bleach, and vinegar shifted urine pH outside the physiological range. Golden-seal tea caused a dark appearance, and specimens containing liquid soap were unusually cloudy. Lemon juice had no effect on the assays. Visine was the only adulterant not detected. The adulterants interfered somewhat differently with each of the drug assays. EIA assays for illicit drugs can be invalidated by specimen adulteration producing false-negative results. Therefore, if urine drug testing is to be conducted, pH, relative density, and appearance should be assessed and suspect specimens should be rejected. Not all adulterants can be detected, so observed collection is strongly recommended. PMID- 3052929 TI - Time-resolved pulsed fluorescence immunometric assays of carcinoembryonic antigen. AB - A time-resolved pulsed fluorescence immunometric assay (TR-PFIA) for carcinoembryonic antigen is described in which either Eu(III) or Tb(III) chelate is used as label. Described in detail is the assay involving the well-documented format of microtiter matrix and Eu(III) fluorescence enhanced with a beta diketone and quantified in a commercial time-resolving fluorometer. We have also used the same basic assay, but one with a Tb(III) chelate as label, and we read the fluorescence signal directly off a surface without the application of enhancement solution. The Tb(III) fluorescence is then brought into solution by using an analog of dipicolinic acid in an enhancement solution. The latter approach demonstrates the scope of the methodology, which invokes the extra complexity of enhancement only when increased sensitivity might be required. The power and versatility of the enhancement methodology are demonstrated. PMID- 3052930 TI - Results compared for tricyclic antidepressants as assayed by liquid chromatography and enzyme immunoassay. AB - The tricyclic antidepressants amitriptyline, nortriptyline, imipramine, and desipramine in serum of patients taking one of the drugs were quantified in two laboratories by high-performance liquid chromatography (HPLC) and enzyme multiplied immunoassay (EMIT; Syva). Results for split samples were highly correlated, but EMIT gave higher results in most cases, and the slopes of the correlation lines for each analyte were greater than 1. Detection limits for the two procedures were such that 18% of the EMIT results for the drug(s) were considered negative, as compared with 4% of the HPLC results. Additional assay of desmethyl or hydroxy antidepressant metabolites by HPLC did not explain the higher EMIT results. The relatively high detection limit for EMIT greatly limits its use in therapeutic drug monitoring, where low concentrations of tricyclic antidepressants are as important as high ones for dose adjustment or determination of compliance. Other problems with EMIT measurement of tricyclic antidepressants are discussed. PMID- 3052931 TI - Clinical and analytical evaluation of two immunoassays for direct measurement of creatine kinase MB with monoclonal anti-CK-MB antibodies. AB - We examined the clinical and analytical performance of two immunoassays (Becton Dickinson CK-MB; Ciba-Corning Magic Lite CK-MB) in which monoclonal anti-CK-MB antibodies are used for directly measuring creatine kinase (EC 2.7.3.2) isoenzyme MB (CK-MB) in serum, and also one electrophoretic method (Ciba-Corning). Within- and between-assay precision for both immunoassays was good at the upper reference limits (less than 10% CV). Analytical recoveries ranged from 102 to 114%. Both immunoassays were free from interference by CK-BB, mitochondrial-CK, macro-CK, adenylate kinase, and CK-MM. Retrospectively, we evaluated four categories of patients, using both immunoassays and electrophoresis: normal controls, acute myocardial infarction (AMI) patients, severe skeletal muscle trauma patients, and acutely ill patients known not to have AMI. In general, there were excellent correlations among all three methods. CK-MB activity (U/L) measured by the Becton Dickinson immunoassay was approximately 50% of the mass concentration (microgram/L) of the Magic Lite immunoassay and 50% of the activity concentration (U/L) determined by electrophoresis. Both immunoassays were easy to perform and sensitive to the low CK-MB concentrations often found with low total-CK activities. PMID- 3052932 TI - Measurement of cortisol with EMIT and Amerlite immunoassays. PMID- 3052933 TI - Intact parathyrin in patients after kidney transplantation. PMID- 3052934 TI - Radioenzymatic assay for simultaneous estimation of dobutamine and endogenous catecholamines. PMID- 3052935 TI - "Tina Quant" immunoturbidometric kit for C-reactive protein adapted to the Cobas FARA centrifugal analyzer. PMID- 3052936 TI - Solid phase immunoassay for high molecular weight alkaline phosphatase in human sera using a specific monoclonal antibody. AB - A murine hybridoma producing monoclonal antibody (MoAb HMAP-1) to human high molecular weight alkaline phosphatase (HMAP) was derived using enzyme partially purified from human serum as the immunogen. The antibody did not cross-react with the liver, bone, intestinal or placental isozymes of alkaline phosphatase (AP), and did not react with AP in bile. A monoclonal antibody immunocatalytic assay for HMAP in serum was developed using MoAb HMAP-1 bound to nitrocellulose membrane discs. The method was rapid and reproducible, giving good correlation with results from cellulose acetate membrane electrophoresis and having the added advantages of increased sensitivity and specificity. A large number of sera can now be assayed to evaluate the significance of serum HMAP in relation to other AP activities and to the differential diagnosis of liver disease. PMID- 3052937 TI - Clinical relevance of increased neuron-specific enolase concentration in cerebrospinal fluid. AB - Neuron-specific enolase (NSE) concentrations in cerebrospinal fluid (CSF) and serum have been studied by an EIA-method using monoclonal antibodies against human NSE. In a control group (n = 24) the mean NSE value (+/- SD) in CSF was 10.8 (+/- 4.5) ng/ml. Increased NSE values in CSF (greater than or equal to 20 ng/ml, ie greater than or equal to means + 2s) have been detected in 33/172 patients with the following neurological diseases: CNS tumors (6/30), infarctions (6/36), cerebral ischemias (5/25), inflammatory diseases (7/33), epilepsias (3/10) and miscellaneous neurological diseases (6/38). The NSE assay in CSF was not specific for a single neurological disease. In 9% of all patients with an organic neurological disease the increased NSE concentration was the only abnormal result in the CNS out of variables routinely determined in CSF. The discrimination between an organic and psychogenic origin of epilepsy may be possible by an NSE analysis in CSF. The NSE assay in CSF can be recommended as an unspecific screening parameter for pathological organic CNS processes. PMID- 3052938 TI - Purification of acidic glutathione S-transferases from human lung, placenta and erythrocyte and the development of a specific radioimmunoassay for their measurement. AB - Human acidic glutathione S-transferases (GST) have been purified from placenta, lung and erythrocytes. The purification protocol resulted in a high yield of pure protein for each tissue, when compared to previous procedures. An apparent subunit Mr of 24,800 was calculated for each of the acidic GST and each enzymes had a pI of 4.75. No immunochemical differences were detected between the acidic GST isolated from the three tissues. A specific and sensitive radioimmunoassay suitable for the measurement of human acidic GST in plasma or tissues is described. PMID- 3052939 TI - The role and techniques of laryngeal electromyography. PMID- 3052940 TI - Fibroblasts of two patients with trisomy 18 show 1.5-fold increase in peptidase A activity over normal human diploid fibroblasts. AB - Peptidase A (Pep A) activity assigned to chromosome 18q23 was biochemically examined in fibroblasts cultured from two patients with trisomy 18 and in fibroblasts derived from normal individuals. The trisomy 18 fibroblasts showed approximately a 1.5-fold increase in Pep A activity over that of the control fibroblasts. Agar gel electrophoretic analysis revealed no detectable differences in the electrophoretic mobility and isoenzyme patterns of Pep A between the trisomy 18 fibroblasts and normal ones. The present results show the trisomy 18 fibroblasts to be suitable for study of the gene dosage effect of Pep A. PMID- 3052941 TI - Specificity of anti-lymphocyte antibodies in sera from patients with AIDS-related complex (ARC) and healthy homosexuals. AB - The presence and specificity of anti-lymphocyte antibodies (ALA) was investigated in sera from male homosexuals with AIDS-Related Complex (ARC) as well as healthy homosexuals. Individuals in the healthy homosexual group had no detectable antibodies to human immunodeficiency virus (HIV). Antibodies reactive with normal peripheral blood mononuclear cells were detected by Western blot analysis in sera from both groups of homosexuals. Of those individuals whose sera contained ALA, 71% of ARC patients and 83% of healthy homosexuals had antibodies recognizing a 73 kilodalton (kD) molecule. ALA present in ARC sera reacted with CD3+, CD4+ and CD8+ lymphocytes while little reactivity with B cells was observed. Our results indicate that ALA appear in homosexuals prior to HIV infection and are reactive primarily with T lymphocytes. A 73 kD structure associated with the T cell membrane is frequently the target for these antibodies. PMID- 3052942 TI - H-Y antiserum recognizes male-specific and non-specific components in human lymphocytes. AB - Serological assays for H-Y antigen, which is male-specific in mammals, all give slightly positive but non-negligible results when females are tested. All assays used to this day share this characteristic; furthermore they do not analyse the pattern of antigens recognized by the antisera. We used the Western blot technique in an attempt to do this, and observed that three components (relative molecular mass: 15-20 kilodaltons (kD)) were recognized both in females and in males tested, and a single specific band of 32-34 kD relative molecular mass in males. This confirms the existence of a serologically detectable male antigen on human lymphocytes, and explains the existence of a certain level of antigen expression in females tested by different techniques, including the indirect immunofluorescence and ELISA used here for comparison. PMID- 3052943 TI - Cell-mediated immunity to pancreatic islet cells in the non-obese diabetic (NOD) mouse: in vitro characterization and time course study. AB - The non-obese diabetic (NOD) mouse is an animal model of insulin-dependent diabetes mellitus (IDDM), in which 80% of the females become diabetic after the age of 12 weeks. Using an in vitro assay we investigated the capacity of spleen lymphocytes from NOD mice to inhibit the insulin secretion of normal islet cells after stimulation by theophylline plus arginine. Spleen cells from diabetic NOD mice inhibited the insulin release of DBA/2 islet cells. Depletion experiments using monoclonal antibodies demonstrated that inhibitory cells belonged to the Lyt2 positive T lymphocyte subset. The phenomenon was not restricted by the MHC class I K region, shared by NOD and DBA/2 mice, since lymphocytes from diabetic NOD mice also inhibited the insulin secretion of normal Wistar rat islet cells. Inhibitory T cells were detected in overtly diabetic mice but also in non diabetic females aged 5-11 weeks indicating that they are not secondary to metabolic disturbances and might contribute to their onset. Conversely they were not found in male NOD mice although some of these mice show insulitis. The presence of these inhibitory T cells might thus represent an early and sensitive marker of anti-islet cell-mediated autoimmunity. PMID- 3052944 TI - Analysis of the spectrotypes of autoantibodies against thyroglobulin in two rat models of autoimmune thyroiditis. AB - We have studied the isoelectric focusing (IEF) spectrotypes of autoantibodies against thyroglobulin in two rat models of Hashimoto's thyroiditis: (1) AUG strain rats immunized with autologous thyroglobulin in Freund's complete adjuvant; (2) PVG/c strain rats which have been thymectomized and sublethally irradiated. The IEF spectrotypes revealed differences between the two models. The anti-thyroglobulin response in immunized AUG rats is mainly oligoclonal or polyclonal, often with dominant clones in the spectrotype, similar to about 90% of Hashimoto's patients, whereas the response in the PVG/c rat is highly restricted. There were pronounced changes in spectrotype with time in the PVG/c, but not AUG, rats with considerable variation in the lifespan of individual clones. The maximum lifespan of an anti-self secreting B-cell clone in the PVG/c rat, determined by persistence of its clonotype, was at least 16 weeks. PMID- 3052945 TI - Adoptive transfer of experimental autoimmune hepatitis in mice--cellular interaction between donor and recipient mice. AB - This report extends our previous study on experimental autoimmune hepatitis in C57BL/6(B6) mice. Cellular immunity involved in the induction of liver injury in this model was studied by transfer of primed spleen cells from hepatitis donor mice to syngeneic normal recipient mice. The most prominent liver damage in recipient B6 mice was induced by transfer of nylon wool adherent spleen cells from hepatitis donor mice, and T cells in this fraction were the essential requirement for the liver damage in the recipient mice. Nylon wool adherent spleen cells from hepatitis donor mice after depletion of the suppressor T-cell function by low-dose (300 rad) irradiation induced more severe liver injury compared to the same cells without irradiation. When the recipient mice were depleted of lymphocytes by low or high dose (700 rad) whole body irradiation, transfer of primed spleen cells from hepatitis donor mice did not induce liver lesion in the lymphocyte-depleted mice. This low susceptibility of lymphocyte depleted recipient mice to primed spleen cells of hepatitis mice was no longer demonstrated after reconstitution with normal spleen cells. In a cell-migration study using 51Cr-labelled spleen cells, it was shown that a considerable number of infiltrating cells in the liver of recipient mice were derived from recipient mice themselves. These results seem to indicate that cell-to-cell interaction between radiosensitive precursor cells of recipient mice and liver-antigen-primed T cells from hepatitis donor mice play an essential role in the induction of liver injury in the recipient mice. PMID- 3052946 TI - Glomerular deposition of food antigens in IgA nephropathy. AB - Recently, we reported on the importance of food antigens on the pathogenesis of an experimentally-induced model of, and some patients with, IgA nephropathy. In this paper, the glomerular deposition of food antigens (casein, lactalbumin, peanut protein, soy bean protein, rice protein, ovalbumin) was investigated by an immunofluorescence technique in 28 patients with IgA nephropathy and 32 controls (ten with lupus nephritis, three with Henoch-Schoenlein purpura nephritis and 19 with other glomerulonephritis). Glomerular IgA deposition was demonstrated in all IgA nephropathy and Henoch-Schoenlein purpura nephritis, and in four lupus nephritis. Positive findings of food antigens, observed as mesangial pattern, were obtained in eleven cases (39.3%) with casein, 21 (75.0%) with soy bean protein and one (3.6%) with rice protein in IgA nephropathy, even though no such findings were seen in the control group. Eleven of 28 patients with IgA nephropathy were positive with soy bean protein alone, nine were positive with soy bean protein + casein, one was positive with soy bean protein + casein + rice protein, and one was positive with casein alone. The deposition of food antigens was not observed in six cases only. Furthermore, no correlation was noted between the deposition of food antigens and the deposition of IgA1, IgA2 or J chain, in vitro binding of the secretory component, or histopathological grades. These results suggest that the exact meanings of glomerular deposition are unclear. Food antigens are postulated, however, as possibly participating strongly in the pathogenesis and as being localized in the glomerular mesangium as an antigen in some patients with IgA nephropathy. PMID- 3052947 TI - Serum IgA preferentially binds to cationic polypeptides in IgA nephropathy. AB - The observation of negatively charged IgA in the mesangium of patients with primary IgA nephropathy (IgA-GN) prompted us to study the charge of serum IgA in IgA-GN, Henoch Schonlein purpura (HSP), alcoholic liver cirrhosis (ALC), membranous nephropathy (MGN) and systemic lupus erythematosus (SLE). Since no abnormal distribution of IgA isoelectric points was detected by isoelectric focusing studies, we developed a sensitive charge-dependent assay using plates coated with either cationized BSA (cBSA) or poly-L-lysine. In 15 IgA-GN sera, the amount of IgA reacting specifically with cBSA (cBSA-IgA) was almost linearly correlated with the poly-L-lysine-binding IgA (r = 0.97, P = 0.0006), suggesting that both assays detect charge-dependent interactions and thus probably measure anionic IgA. Significantly high serum levels of cBSA-IgA were observed in 56% of IgA-GN patients and in 40% of ALC patients. In contrast, normal serum levels of cBSA-IgA were detected in HSP, MGN and SLE. Both, the mono- or polymeric IgA bound to cBSA in a patient's serum studied. Contrasting with the presence of anionic IgA, no increase of cBSA-IgG was observed in IgA-GN. IgA rheumatoid factor (IgA-RF) assay showed high levels in IgA-GN (39%) and in ALC (25%). IgA-RF levels did not correlate with the amount of cBSA-IgA. When 18 patients with IgA GN were tested after kidney transplantation, increased levels of cBSA-IgA and/or IgA-RF were found to be associated with the recurrence of mesangial IgA deposits in the graft. This suggests that both negatively charged IgA and IgA-RF may play a role in the recurrence of mesangial IgA deposits. PMID- 3052948 TI - Immunosuppression induced by talc granulomatosis in the rat. AB - Granulomatosis caused by four subcutaneous talc powder-suspension injections induced strong immunosuppression in rats. The disturbance included reduction of mononuclear white blood cell count in the peripheral blood, atrophy of the thymic cortex, spleen enlargement with predominance of red over the white pulp, increase in the number of lymph node germinal centres and a significant delay of the first set and second-set allograft rejection. Neither phagocytic function of reticuloendothelial system nor erythrocyte count and humoral immune response were found to be altered. Indomethacin suppression of prostaglandin production did not normalize the allograft rejection dynamics. In contrast, splenectomy completely abolished the immunosuppressive effects of granulomatosis. In splenectomized, talc-treated animals WBC counts were not altered and the rejection of allografts was not delayed. Suppression of immune response to alloantigens was transferred to normal and splenectomized recipients by both serum and spleen cells of talc injected animals. Also, in a cell mixture-transfer experiment, spleen cells from talc-granulomatosis-bearing donors suppressed the immune response induced by lymph node cells from immune donors in T cell-deficient rats. The inability of serum from splenectomized talc-injected rats to transfer the suppression suggested the crucial role of the spleen in the mechanisms leading to suppression in rats bearing talc-granulomatosis. PMID- 3052950 TI - Percutaneous renal biopsy well-visualized by orthogonal ultrasound application using linear scanning. AB - We attempted 24 percutaneous renal biopsies under orthogonal ultrasonic guidance in 23 children with renal diseases aged 1 to 17 years old. We used a 5 MHz linear array transducer which was positioned on the side of the abdomen and aligned parallel to the biopsy needle so that projected ultrasound reaches the needle orthogonally. Thus, we could clearly and continuously monitor the biopsy needle and transverse plane of the kidney during the biopsy. We obtained diagnostically sufficient renal specimens in all subjects and found no significant complications, including macroscopic hematuria, in any case. We found that our method simultaneously visualized the biopsy needle and kidney much more clearly and made the biopsy far easier and safer than any other method previously proposed and we recommend the procedure be widely used in adult as well as child patients. PMID- 3052949 TI - Immunohistochemical identification of heparan sulphate proteoglycan in secondary systemic amyloidosis. AB - The distribution of proteoglycans in kidneys from patients with secondary (AA) systemic amyloidosis was investigated. Antisera reacting with the protein cores of chondroitin sulphate proteoglycan (CSPG), dermatan sulphate proteoglycan (DSPG) and heparan sulphate proteoglycan (HSPG) were used in conjunction with the peroxidase-antiperoxidase (PAP) method. HSPG was the only proteoglycan found to be specifically localized to the amyloid deposits. The staining was most intense on the endothelial side of the deposits in both the glomeruli and in the vessel walls. No staining was observed after absorption of the HSPG antiserum with a fraction of the amyloid preparations, corresponding in size to that reported for glomerular HSPG. The possible role of HSPG and endothelial cells in the pathogenesis of the amyloid deposits is discussed. PMID- 3052951 TI - Anti-glomerular basement membrane antibody and linear glomerular immunofluorescence in a patient with systemic lupus erythematosus. PMID- 3052952 TI - Congenital renal arteriovenous fistula. PMID- 3052953 TI - Carcinogenic glutamic acid pyrolysis product in the dialysate of uremic patients treated by continuous ambulatory peritoneal dialysis. AB - By using a high-performance liquid chromatography (HPLC) method, we determined the contents of 2-amino-6-methyldipyrido[1,2-a:3', 2'-d] imidazole (Glu-P-1) and 2-aminodipyrido[1,2-a:3', 2'-d]imidazole (Glu-P-2) in the dialysate of patients with uremia. The total dialysate (approximately 6 liters) per day of the patients who had received continuous ambulatory peritoneal dialysis (CAPD) was examined. The amounts of Glu-P-1 and Glu-P-2 in the total dialysate per day were 552.2 +/- 266.9 pmole and 386.3 +/- 146.0 pmole (mean +/- SD, n = 10), respectively. The recoveries of Glu-P-1 and Glu-P-2 in the dialysate on our assay method were 68.2% and 54.3%, respectively. Based on the recoveries, the average amounts of Glu-P-1 and Glu-P-2 in the dialysate of uremic patients were assumed to be 809.7 pmole and 711.4 pmole, respectively. PMID- 3052954 TI - Intra-arterial digital subtraction angiography for children with idiopathic renal bleeding: a diagnosis of nutcracker phenomenon. AB - Among children with asymptomatic hematuria, 28 cases of nonglomerular idiopathic renal bleeding were subjected to this series of study. Intra-arterial digital subtraction angiography (DSA) and/or renal venography were performed to investigate the hematuria of unknown etiology. DSA clearly demonstrated the entrapment of the left renal vein (LRV), or nutcracker phenomenon in the majority of our patients (22 out of 28 cases): obstruction of the LRV with well-developed collaterals were found in 8 cases, and in the remaining 14 cases, various degrees of LRV compression were demonstrated. A characteristic real-time DSA image was the congestion of LRV associated with collaterals and/or intermittent venous flow at the compressed segment of LRV. The pullback pressures from LRV to the inferior vena cava (IVC) that were obtained from 5 of these patients demonstrated gradients of 2 mmHg (3 cases), 3 mmHg (1 case), and 5 mmHg (1 case), respectively. The parallel application of ultrasonography has given positive signs for LRV entrapment, although they have not necessarily coincided with the existing criteria of nutcracker phenomenon. Considering the high incidence of LRV entrapment among children with nonglomerular hematuria, most nutcracker phenomenon should be diagnosed on ultrasonography. However, intra-arterial DSA is an important tool to establish the disease entity and ultrasonic criteria. PMID- 3052955 TI - Detection of novel beta 2-microglobulin in the serum of hemodialysis patients and its amyloidogenic predisposition. AB - Serum from 8 undialyzed patients and 30 dialyzed patients was examined by immunoblotting using anti beta 2-microglobulin (beta 2M) serum after two dimensional gel electrophoresis (2.DE). One major spot and three minor spots were detected in the ultrafiltrate as well as in the serum. One major spot was determined to be native beta 2M and three minor spots were found to be novel beta 2M. Novel beta 2M had a lower molecular weight (MW) and a higher acidic isoelectric point (pI). Novel beta 2M was recognized in the sera of 5 out of 20 hemodialysis (HD) patients without carpal tunnel syndrome (CTS), 2 of whom had been on HD from 5 to 10 years and 3 for more than 10 years, as well as in the sera of all 10 patients with CTS. By chromatofocusing, pI of novel beta 2M was 5.2, while pI of native beta 2M was 5.7. When the tissue specimen of transverse carpal ligament of 2 HD patients with CTS was examined by immunoblotting after 2.DE, the spot of novel beta 2M was larger than that of native beta 2M. It is possible that some metabolic abnormality of beta 2M occurs through long-term hemodialysis, and it is possible that novel beta 2M might relate to amyloidogenic predisposition. PMID- 3052956 TI - Gastrointestinal involvement of dialysis-related amyloidosis. AB - Beta-2 microglobulin-related amyloids were found in gastric biopsy specimens from 3 of 11 patients who had been on hemodialysis more than 10 years and examined. The deposits were found not only in arteriolar walls but also in lamina propria and muscle tissue. In another patient who was on hemodialysis for 8 years, beta 2 microglobulin-related amyloids were found in the muscle layer of operatively resected colon. These four patients also had the same substance deposited in the osteoarticular system. The results suggest the systemic involvement of beta 2 microglobulin-related amyloidosis in patients on long-term hemodialysis. PMID- 3052957 TI - CAPD and transplantation in diabetics. AB - Treatment of diabetics with end-stage renal failure is still a hazard. We report on 33 insulin-dependent diabetic patients who were treated with CAPD over a period of up to 70 months. Seven of them have been transplanted. The evaluation of the clinical and biological parameters serum protein, phosphate, calcium, revealed acceptable values. Hemoglobin rose during the treatment period. Hypertension improved and medication was reduced. HbAlc levels were near normal with subcutaneous application of insulin. Peritonitis rate was 1/18 patient months. Survival rate was comparable with other centers with 83% in the first and 62% in the second year. In 7 patients that have been transplanted no CAPD-related problems arose after transplantation. 6 of them are still doing well; one returned to CAPD because of graft failure. In conclusion, because of good control of hypertension, blood glucose and anemia and the avoidance of fluctuating volumes, we think CAPD to be the best method for diabetics awaiting transplantation and for those that cannot be transplanted. PMID- 3052958 TI - Pharmacological manipulation of peritoneal transport in CAPD. AB - Various endogenous compounds and drugs influence different determinants of peritoneal transport. Catecholamines affect vascular diameter and mesenteric blood flow. Several gastrointestinal hormones were found to induce splanchnic vasodilation and to augment the effective vascular surface. Different prostanoids, histamine, and bradykinin influence peritoneal mass transfer by increasing vascular permeability. Vasodilative acting drugs, diuretics and surface active compounds were found to augment peritoneal solute clearances and water efflux by acting on peritoneal vessels or the serosal surface of the peritoneal membrane. Hyperosmolar dialysis solutions increase ultrafiltration and contribute to total peritoneal clearance by convective transport. The present paper aims to summarize the current knowledge on pharmacological and physiological manipulation of peritoneal transport function and to show some ways of its clinical use. PMID- 3052959 TI - The immune status of the uremic patient: hemodialysis vs CAPD. AB - The immune status of the uremic patient is characterised by an affected cellular immunity. The absolute numbers of circulating T-cells are reduced. The relative numbers of OKT8-cells and the suppressor cell activity seem to be increased, the helper cell activity seems to be decreased. Hemodialysis does not restore the impaired immune status. With increasing time of dialysis the alterations become more pronounced. So far a difference in the immune status of CAPD patients compared to HD patients has not been proven. The impairment of immune response might be less than in HD patients. The mechanism of this difference is not established. However, the question of immune response in uremia and its alteration by different forms of dialysis treatment is only in part and insufficiently investigated. Studies in CAPD patients have only a very preliminary character. PMID- 3052960 TI - Immunologic patterns in CAPD patients with peritonitis. AB - Alterations of peritoneal defense mechanisms, i.e. opsonization, phagocytosis, and bacterial killing, may be responsible for the high peritonitis rate in a subgroup of CAPD patients. Peritonitis incidence and in vitro bacterial opsonization has been shown to depend on IgG concentrations in the dialysate and on the ability of macrophages to produce fibronectin. Additionally, a high peritonitis incidence is associated with decreased bactericidal activity of macrophages; thus infective organisms may survive intracellularly despite intact phagocytosis. In some CAPD patients a disturbance in lymphokine and monokine release may be responsible for the reduced ability of their macrophages to kill bacteria. PMID- 3052961 TI - Effects of long-term CAPD on carbohydrate and lipid metabolism. AB - Long-term follow-up of CAPD patients indicates that these patients are at relatively low risk for developing de novo diabetes mellitus. Transperitoneal glucose absorption and the blood glucose and serum insulin response to the glucose load remain relatively unchanged with time. Alterations in lipid metabolism do occur; there is a tendency for the triglyceride levels to increase after 1 year and to maintain these high values with time. Total HDL cholesterol and its subfractions HDL2 and HDL3 remain unchanged and in the low normal range. Apoprotein A1 levels increase after 1 year and are maintained in the normal range. PMID- 3052962 TI - Renal osteodystrophy and aluminum bone disease in CAPD patients. AB - Renal failure is frequently associated with osteodystrophia due to secondary hyperparathyreoidism and/or increased aluminum intake. The problem of hypercalcemia and hyperphosphatemia can more easily controlled by CAPD than by hemodialysis. Total serum and ionized calcium levels are rapidly normalized by a CAPD regime of four 2-1 exchanges with 1.75 mmol/l Ca. Under the same CAPD regime 250-300 mg phosphate are removed per day. Depending on the ingestion of phosphate, 100-200 mg phosphate per day remain to be removed by phosphate binding agents. Since the main source of aluminum in CAPD patients is oral ingestion of aluminum-containing phosphate binders, serum levels should be regulated by diet and calcium carbonate. To suppress PTH secretion serum ionized calcium levels need to be maintained at the upper limit of normal. This can also be achieved by the use of oral calcium carbonate. Vitamin D or analogs should be prescribed only when clinically indicated by persistent hypocalcemia, osteitis fibrosa or non aluminum related osteomalacia. PMID- 3052963 TI - CAPD and systemic diseases. AB - A review is presented on the use of continuous ambulatory peritoneal dialysis (CAPD) in some systemic diseases with renal involvement to the stage of terminal renal failure: progressive systemic sclerosis, systemic lupus erythematosus, paraproteinemias and systemic amyloidosis. CAPD provides a suitable means of treatment of end-stage renal failure complicating systemic diseases both in terms of control of uremia and quality of life, being the treatment of choice in selected patients. PMID- 3052965 TI - A crossover study of naproxen, diclofenac and tolmetin in seronegative juvenile chronic arthritis. AB - Nonsteroidal anti-inflammatory drugs (NSAID) are useful in the management of juvenile chronic arthritis (JCA) and efficacy has usually been compared with aspirin. The disadvantages of aspirin include the frequency of dosage, monitoring of salicylate levels and concern about Reye's syndrome. We have performed a single blind crossover study comparing naproxen 10 mg/kg/day, tolmetin 25 mg/kg/day and diclofenac 2 mg/kg/day in 28 children with seronegative JCA over 12 weeks. Clinical and laboratory assessments were made after 4 weeks of treatment on each drug. Clinical improvement was seen with all three NSAID compared to baseline after a 24-hour washout period, but statistical significance was reached in only 2 or 3 of the clinical parameters for each drug. Side effects were mild and typical of NSAID, but occurred less frequently with naproxen (5) and tolmetin (5) than with diclofenac (7). Thus the anti-inflammatory effect of all three drugs was confirmed with no significant differences between them in terms of efficacy and tolerance. Tolmetin 200 mg tablets are now only available in France and N. America, therefore naproxen or diclofenac are recommended as the first line of treatment in children with chronic arthritis over the age of 5. PMID- 3052964 TI - Anti-idiotypic antibodies in autoimmune diseases. AB - The variable region (V region) of an antibody, the part of the molecule that binds to antigen, is itself antigenic and can elicit anti-V region antibodies when injected into animals of a different or even of the same species. The antigens of the V region constitute its idiotype, and they are defined by anti idiotypic antibodies in polyclonal antisera or by monoclonal anti-idiotypic antibodies. An idiotype actually consists of multiple antigenic determinants, each of which is an idiotope. The antigenic determinants or idiotopes can reside in the heavy chain component of the V region, in its light chain component, or they may consist of a surface made up of parts of both chains. The idiotype of an antibody is, therefore, a way to describe by serological means the variable region of an antibody molecule. Originally, idiotypes were defined by heterologous antisera made, for example, by immunizing a rabbit with a monoclonal human myeloma protein. After extensive absorption with normal human immunoglobulins, such antisera were found to bind only to the V region of the immunizing myeloma immunoglobulin. Thus, idiotypes were originally thought to be unique markers of individual antibodies (hence their name). However, it is now known that different antibodies can share the same, or similar, idiotype. Even antibodies with different antigen-binding specificities can share the same idiotype if they use the same heavy or light chain in forming their V regions. Such antibodies constitute parallel sets. Thus, idiotypes can be: private (confined to a particular immunoglobulin molecule), public (shared by different antibodies), or cross-reactive (different idiotypes containing similar antigenic structures).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3052966 TI - Rheumatic diseases and pregnancy: a perspective. AB - The interaction between certain rheumatic diseases, sex hormones and their wide fluctuation during pregnancy and postpartum may be responsible for the variable course of rheumatic diseases during pregnancy. Important issues include effects on the mother and fetus by the disease, pregnancy and maternal drug therapy. PMID- 3052967 TI - Ankylosing spondylitis and pregnancy. AB - The pregnancy-induced remission typical of RA does not seem to occur in pregnant women with AS. The mechanisms for this difference are not understood, but appear to be related to different pathogenetic mechanisms operating in the two diseases. The vast majority of women with ankylosing spondylitis can expect to have the same rate of fertility, course of pregnancy and birth, and to give birth to normal healthy babies to the same extent as the normal female population. The chance for the offspring to contract AS later in life is somewhat increased. PMID- 3052968 TI - The neonatal lupus syndrome. AB - The neonatal lupus syndrome and congenital heart block are strongly associated with the presence of antibodies to the ribonucleoprotein antigen Ro. The study of these conditions has given insight into possible pathogenetic mechanisms operating in connective tissue diseases. PMID- 3052969 TI - Pregnancy in mixed connective tissue disease, poly/dermatomyositis and scleroderma. AB - Pregnancy related problems in mixed connective tissue disease, polydermatomyositis and scleroderma are analysed. Particular attention is also elevated to the therapeutical approach to these diseases during pregnancy and delivery. PMID- 3052970 TI - SLE and pregnancy. AB - In patients with SLE in remission pregnancy does not confer a special risk to either the mother or foetus, and immunoregulatory drugs should not be withdrawn prior to a pregnancy where these are required for disease control. Likewise therapy to treat disease flares during pregnancy should not be withheld as the foetus is at comparitively more risk from the effects of the maternal disease than from the small risk associated with therapy. Patients with active disease should be counselled against pregnancy but therapeutic abortion in such patients is only very rarely indicated. High risk groups benefit from close monitoring though the previous obstetric history will not necessarily reflect outcome in subsequent pregnancies. Patients without a previous obstetric history should not be treated prophylactically due to the detection of antiphospholipid antibodies, and the optimum treatment for those with poor histories and aPL is currently being investigated. PMID- 3052972 TI - Colchicine in therapy. State of the art and new perspectives for an old drug. AB - Colchicine is the most specific treatment in acute gouty attacks. In several European countries, oral colchicine is still used for routine treatment of acute gout. Its selectivity is used as a diagnostic tool. It is also active in the treatment of acute crises of chondrocalcinosis and more occasionally of other arthritic crises (e.g. sarcoidosis). Colchicine appears to be the necessary adjuvant prophylactic drug when starting a hypouricemic treatment with uricosuric or uricolytic drugs for avoiding acute gouty crisis due to sudden mobilisation of the uric acid pool. Besides gout, colchicine is the drug of choice for treating familial mediterranean fever. It appears to be helpful in the treatment of Behcet's disease. It seems also useful for treating fibrosing conditions such as liver cirrhosis and scleroderma. As an adjuvant therapy, it helps treating dermatological disorders which are associated with leucocyte migration as an essential pathogenic factor (e.g. psoriasis, dermatitis herpetiformis, necrotising vasculitis ...). It has been advocated as an adjuvant therapy in malignant diseases as a support in radiotherapy and as an useful drug in various other diseases where it has been tried occasionally (e.g. Paget's disease of the bone, idiopathic thrombocytopenic purpura, disc syndrome). This very old drug remains a modern therapeutic agent. PMID- 3052971 TI - The effect of physical training on patients with rheumatoid arthritis: changes in disease activity, muscle strength and aerobic capacity. A clinically controlled minimized cross-over study. AB - For decades, physical training of rheumatoid arthritis (RA)-patients has been controversial, especially for patients with active disease. The aim of this study was to investigate whether RA-patients could receive graduated training without increasing the activity of the disease. In a controlled cross-over study the effect of graduated progressive training has been evaluated in 18 RA-patients with moderately active disease. The training was performed twice weekly with aerobic conditioning and strength exercises progressing to strenuous exercises over an 8-week period. The design was a crossover project with two groups obtained by minimisation. After training the patients had significantly fewer swollen joints than before. Training of the muscles acting over the swollen joints resulted in more than a 35% decrease in the number of swollen joints. The hemoglobin level increased significantly after the training period. The erythrocyte sedimentation rate, the complement factor C3d, and the number of sore joints remained unchanged. A decrease in the need for medicine was non significant. From this study it appears that RA-patients with some activity are trainable without aggravating the disease, even in the chronically swollen joints. The rheumatoid arthritis activity decreased with fewer swollen joints and higher hemoglobin level after training. PMID- 3052973 TI - Differential effect of inflammatory stimuli on murine plasma C4 and factor B concentrations. AB - The in vivo effects of a variety of inflammatory stimuli on complement C4 and factor B plasma levels have been examined. MRL/++ (H-2k) mice were given intraperitoneal injections of lipopolysaccharide, turpentine, Corynebacterium parvum pyridine extract residue or high doses of indomethacin. All of these treatments induced an increase in plasma factor B concentrations, which in the case of C. parvum was dose dependent and persisted for at least 7 days. Lipopolysaccharide, turpentine and indomethacin produced decreases in plasma complement C4. C. parvum, however, produced an increase in plasma complement C4 to approximately 240% of controls which was independent of gender. It was also independent of major histocompatibility complex haplotype, since the same effect was seen in C57B1/6J-bg/bg and C57B1/6J-bg/+ mice. The gross increment in complement C4 was, however, related to the major histocompatibility complex. H-2K mice ("low complement C4") had smaller increments than H-2b ("high complement C4"). Mycobacterium bovis (BCG) also produced a transient increase in C4 in the H 2b mice as well as a prolonged increase in factor B levels. These data (i) suggest that different inflammatory stimuli induce different mediators which may have differential effects on factor B and complement C4 synthesis, and (ii) emphasize the independent regulation of complement C4 and factor B. Qualitative variations in the mediators elaborated during chronic inflammatory diseases may help determine complement C4 fluctuations in systemic lupus erythematosus and the wide range of complement C4 concentrations seen in MRL/1 pr mice with active immune complex disease. PMID- 3052974 TI - Failure of methylprednisolone acetate to prolong the antinauseant effect of intravenous methylprednisolone sodium succinate in patients receiving chemotherapy. AB - The efficacy of intramuscular methylprednisolone acetate in maintaining the antinauseant effect of intravenous methylprednisolone sodium succinate was assessed in a prospectively-randomized, double-blind, crossover-design trial. Of 150 patients entered, 127 were evaluable. There was no statistically significant difference between methylprednisolone acetate and saline placebo, although patient preferences slightly favoured the methylprednisolone acetate for nausea, vomiting, and overall effectiveness. PMID- 3052975 TI - A new osteotomy for cubitus varus. AB - From 1974 to 1986, a step-cut technique of distal humerus valgus osteotomy using one cortical screw for fixation was used to correct cubitus varus deformity in 11 patients. The results were graded as excellent, eight patients, good, two, and poor, one. The poor result was secondary to persistent varus. The average humeral elbow-wrist angle in the ten patients with valgus correction roentgenographically measured 9.3 degrees. The average amount of correction was 28.4 degrees. All patients retained their preoperative level of elbow motion. There were no radial or ulnar nerve injuries, nonunions, infections, or hypertrophic scars. The osteotomy requires careful preoperative planning and special attention to surgical detail. Large amounts of deformity may be corrected safely with a low complication rate. PMID- 3052976 TI - Heparinized materials for control of the formation of the laminectomy membrane in experimental laminectomies in dogs. AB - This study examined the efficacy of heparinized biodegradable materials in the prevention of post-laminectomy scar formation using a canine system. Six-level noncontiguous laminectomies were performed on five adult canines. Exposed dura at each level was covered with the following: (1) nothing (control); (2) fat; (3) fat and autogeneic bone; (4) heparinized Avitene; (5) heparinized Surgicel; or (6) heparinized collagen gel. Canines were killed 16 weeks postoperatively and histologic specimens examined. Fibrous adhesions were abundant at Site 1 (control). Free fat grafts were well preserved and did not permit adhesions at Sites 2 and 3. None of the heparinized materials was as effective as fat in controlling scar formation. Overlying bone at Site 3 did not alter the quality of a free fat graft placed over the dura, a point possibly significant for laminectomies followed by fusion. PMID- 3052977 TI - Repair of segmental defects of the tibia with cancellous bone grafts augmented with human bone morphogenetic protein. A preliminary report. AB - Human bone morphogenetic protein (hBMP) is a bone cell differentiation-inducing factor. Six patients with traumatic segmental three- to 17-cm tibial defects developed solid union by implantation of hBMP and autogeneic cancellous grafts and stabilization. There were no allergic, infectious, or surgical complications. If hBMP augmentation in biodegradable delivery systems can be established by a prospective, randomized, double-blind investigation, the incidence of successful bone graft operations for treatment of large segmental defects would be measurably improved. PMID- 3052978 TI - Traumatic separation of epiphyses. An experimental study in rats. AB - Four types of physeal fracture-separations, defined by the Salter-Harris classification, were created experimentally in the proximal physis of the right tibia of immature rats. The four types are: Type I, pure epiphyseal separation; Type II, separation of the epiphysis with a metaphyseal fracture; Type III, partial epiphyseal separation with a vertical fracture of the epiphysis; and Type IV, vertical epiphyseal and metaphyseal fractures. A sham operation performed on the left tibia served as the control. The animals were killed at various intervals up to 25 days after the operation. The findings were assessed by roentgenographic, histologic, and vessel injection methods. For Types I and II morphometric analyses were also performed. The influence of various types of lesions on the growth rate and the healing process was documented. For Types I and II lesions there was a transitory growth arrest and an increased thickening of the zone of hypertrophic cells. The alterations regressed after Day 15, and by day 25, a nearly normal plate was seen. For Type III lesions an angular deformity of the tibia occurred that increased with time. For Type IV lesions, a step-off developed on the articular surface that became more severe with time. Early vascular anastomoses between the epiphysis and the metaphysis led to the formation of bone bridges in Type III and Type IV lesions. PMID- 3052979 TI - Articular-plate replacement arthroplasty for the knee joint. 1964. PMID- 3052980 TI - Magnetic resonance imaging in the diagnosis of aggressive villonodular synovitis. AB - An aggressive and recurrent pigmented villonodular synovitis of the knee occurred in a 47-year-old man. Magnetic resonance imaging was an excellent noninvasive diagnostic aid that completely outlined the extent of the synovitis and correlated well with surgical findings. PMID- 3052981 TI - Bone graft and electrical stimulations during fracture repair. PMID- 3052982 TI - The significance of poor renal transplant concentrating ability as assessed by I 131 Hippuran imaging. AB - A pattern of very poor renal allograft concentration of I-131 Hippuran and good bladder activity has been observed by the authors. The kidneys are typically barely visualized and the renogram curve is flat. The significance of this particular finding has not been described in the literature. Accordingly, a retrospective review of all patients exhibiting this scan pattern was performed and the authors attempted to correlate it with clinical, laboratory, and pathological findings at the time of study. Of 11 patients exhibiting this scan pattern, three had very high urine outputs at the time of study. Of the remaining eight patients, five had chronic rejection. In the remaining three patients, the diagnosis was: one each, acute rejection, mixed acute and chronic rejection, and cyclosporine toxicity. The authors conclude that although this pattern is non specific, if the patient is not in a high output state, it is usually related to chronic rejection. PMID- 3052983 TI - Delayed accumulation (flip flop) of Tc-99m DTPA in a leiomyosarcoma. AB - An 85-year-old woman with leiomyosarcoma occupying the abdominopelvic area had a renal dynamic study with Tc-99m DTPA. The tumor area initially showed almost no content of the radiopharmaceutical. At 24 hours, there was retention of activity in the leiomyosarcoma as compared with surrounding tissues (flip flop). It was noted that, based on literature reports, tumors of mesodermal origin (leiomyoma, spindle cell sarcoma, neurofibroma) may have the ability to concentrate Tc-99m DTPA. PMID- 3052989 TI - Mercury poisoning in a 19th century naval vessel. PMID- 3052988 TI - Dental branch practices in the early 19th century. PMID- 3052986 TI - Clinical pharmacokinetics of drugs used in the treatment of breast cancer. AB - This article reviews the absorption, bioavailability, distribution, metabolism, and elimination patterns of the agents most commonly used for the treatment of breast cancer. Although the majority of the pharmacokinetic studies reviewed have been examined in non-breast cancer patients, the kinetic findings are not significantly altered by disease state. The anthracyclines, antimetabolites, alkylating agents, antioestrogens and mitotic inhibitors are among the classes of agents used to treat breast cancer. Although extensively examined, cancer pharmacokinetic research has resulted in very few clinically relevant findings in breast cancer. PMID- 3052990 TI - A city set on a hill: the story of dentistry in Dundee up to 1940. PMID- 3052987 TI - Inhibition of drug metabolism by quinolone antibiotics. AB - A number of quinolone antibiotics have been found to reduce the hepatic clearance of coadministered drugs such as theophylline. Enoxacin appears to be the most potent inhibitor, consistently decreasing theophylline clearance by more than 50%, while a single study suggests a similar degree of inhibition with pipemidic acid. Ciprofloxacin and pefloxacin reduce theophylline clearance to a smaller extent (approximately 20 to 30%). However, with ciprofloxacin, larger changes and theophylline toxicity have been reported in some subjects. Norfloxacin, ofloxacin and nalidixic acid appear to have minimal effects on theophylline clearance. Enoxacin and ciprofloxacin have also been found to reduce the clearance of caffeine, while ofloxacin has no effect. Few other substrates have been studied. Enoxacin decreases the clearance of R-warfarin with no effect on S-warfarin. In addition, enoxacin has been reported to reduce the clearance of antipyrine, with no effect on chlorpropamide, glibenclamide (glyburide) or phenytoin. The mechanism of these interactions is largely unexplored. It has been suggested that inhibition may be related to the production of 4-oxoquinolone metabolites; however, this hypothesis has not been confirmed. No unique structural feature has been identified to date which explains differences between these compounds in their propensity to affect drug metabolism. Further studies are needed to evaluate the effects of these drugs on other substrates not yet examined and to assess whether or not inhibition is dose related. Clinically, caution is advised when using a quinolone, particularly enoxacin, pipemidic acid, ciprofloxacin or pefloxacin, in combination with theophylline. Close monitoring of theophylline concentrations is recommended in any patient receiving these drugs. The clinical significance of inhibited metabolism of other substrates remains unclear at present. Until further data are available, clinicians should be aware of the possibility of reduced drug clearance resulting in adverse effects whenever the fluoroquinolones are coadministered with drugs that depend on hepatic metabolism for their elimination. PMID- 3052985 TI - Digitalis. An update of clinical pharmacokinetics, therapeutic monitoring techniques and treatment recommendations. AB - The intestinal absorption of digoxin is essentially a passive non-saturable diffusion process, although a saturable carrier-mediated component also plays an important role. The bioavailability varies between 40 and 100%: the presence of food may reduce the peak serum concentration, but does not reduce the amount of digoxin absorbed. Recent development of a capsule containing a hydroalcoholic vehicle may reduce interindividual variations in absorption. Pharmacokinetic analysis of the distribution of digoxin suggests 3 compartments, the slow distribution phase accounting for the lag time between the inotropic effects and the plasma concentration profile. Digoxin is extensively bound to tissues such as myocardium, renal, skeletal muscle as well as red blood cells, but not to adipose tissue. Plasma protein binding varies between 20 and 30%: displacement of digoxin from protein binding sites does not cause significant clinical effects. As expected, haemodialysis or exchange transfusions do not significantly alter the body load of digoxin. The apparent volume of distribution of digoxin varies between 5 and 7.3 L/kg; this may be reduced by, for example, electrolyte abnormalities which reduce digoxin binding to the myocardium. The elimination half-life of digoxin is 36 hours, with 60 to 80% being excreted unchanged, by passive glomerular filtration and active tubular secretion. The remainder is excreted non-renally. Clearance is therefore dependent on renal function and declines in renal disease and in elderly patients. Digoxin interacts with other drugs at any stage of absorption (e.g. cholestyramine), distribution (e.g. quinidine), metabolism (e.g. phenytoin) or elimination (e.g. diltiazem). Patients should, therefore, be carefully monitored when changing a therapeutic regimen which includes any drugs known to interact with digoxin. Clinical monitoring is more important than therapeutic drug monitoring which should be reserved for suspected toxicity, doubts about efficacy, or in cases of poor compliance. With the advent of newer treatment modalities, digoxin is no longer the treatment of first choice in supraventricular arrhythmias and congestive heart failure. However, with careful monitoring, digoxin remains an important therapeutic option. PMID- 3052991 TI - [ECG-triggered magnetic resonance tomographic measurement of blood flow velocity in the carotid arteries: comparison with duplex sonography]. AB - Blood flow velocity in the common carotid arteries was measured for 30 vessels by MRI with a multiecho SE-sequence and compared with the results from duplex ultrasound. MR blood flow velocity was measured for a time interval of 390 ms every 30 ms using a EKG-triggered data acquisition. The time-velocity curves measured by MRI and duplex ultrasound were in good agreement. The correlation coefficient for the peak velocity was r = 0.83. Mean peak velocity as measured by MRI was 59 +/- 8% of peak velocity as measured by duplex ultrasound. PMID- 3052992 TI - The abdominal manifestation of von Hippel-Lindau disease and a radiological screening protocol for an affected family. AB - The mortality in von Hippel-Lindau disease (VHL) is due to the development of central nervous system or abdominal malignancies, particularly renal cell carcinomas. The abdominal manifestations of VHL discovered in the radiological screening of 22 members of an affected family are described. We discuss the problems of making the diagnosis and of detecting small renal and pancreatic tumours against a background of multiple cystic disease. A radiological screening and surveillance protocol using ultrasound and computed tomography is described. PMID- 3052993 TI - Ultrasound changes in the transplant kidney. AB - The purpose of this trial was to study the ultrasound changes that take place in a non-selected group of transplant kidneys in the post-operative period and to identify the ultrasound signs that most accurately identify episodes of acute rejection. It was found that in patients with no episodes of rejection or acute tubular necrosis the ultrasound appearances of the kidney progressed in a predictable way to reach a stable picture similar to the non-transplanted kidney after 2 weeks. Those patients who had episodes of early rejection, but subsequently 'settled down' to a stable state had a similar progress between rejection episodes but the ultimate stable appearance of the kidney varied greatly. Those patients with recurrent rejection episodes, leading to eventual failure of the transplanted kidney, had an unpredictable sequence of ultrasound changes. These findings demonstrate that early rejection episodes may permanently affect the appearance of the kidney and this makes interpretation of future scans difficult unless a baseline scan is available for comparison. The comparison of cortico-medullary echogenicity with a baseline scan makes ultrasound a more specific test than previously reported, but does not improve the poor sensitivity. PMID- 3052994 TI - Ultrasound-guided fine-needle histological biopsies in the abdomen. AB - The results of 83 consecutive ultrasound-guided abdominal fine-needle biopsies are reviewed and the technique described. Material suitable for histological examination was obtained in 87% of cases. The overall sensitivity for the diagnosis of malignancy was 82% and the overall accuracy was 86%. The predictive value of a positive result was 100%. Of the biopsies positive for malignancy, 61% provided further descriptive histological information, usually allowing a differential diagnosis, and in 35% of the positive biopsies a specific histogenesis or diagnosis was suggested. There were no complications with this procedure. This technique was found to be of value for the diagnosis of abdominal lesions and is recommended where there is no cytology service available. The technique has certain advantages over aspiration cytology and could prove to be superior. PMID- 3052995 TI - Assessment of portal vein patency: comparison of arterial portography and ultrasound scanning. AB - The accuracy of ultrasound assessment of portal vein patency has been defined by comparing it with the results of arterial portography in 115 cases. The accuracy of arterial portography was confirmed in 21 cases where orthotopic liver transplantation was performed and used as a 'bench-mark' against which to assess the ultrasound findings. Ultrasound correctly assessed portal vein patency in 87.5% of patients. It was more accurate in assessing patency (90%) than occlusion (68%). Ultrasound correctly assessed portal vein patency in 90% of cases of cirrhosis and hepatic malignancy. Difficulties occurred in children with biliary atresia particularly following the Kasai operation (37.5% accuracy). In the absence of previous surgery to the portal vein or biliary system, ultrasound is comparable to arterial portography and can be used as the sole means of assessment. PMID- 3052996 TI - The sonographic features and implications of fetal pleural effusions. AB - Pleural fluid was detected by ultrasound in three fetuses as an anechoic area surrounding the echogenic fetal lung. In one fetus the underlying lung was small, immobile and of abnormal contour; death occurred in the neonatal period and the lung was found to be hypoplastic. In two other fetuses the lungs appeared sonographically normal, being mobile with normal contours. Both survived, one following spontaneous intra-uterine resolution of the effusion and the other after early neonatal intervention prompted by the antenatal diagnosis. Sonography can detect pleural fluid in utero and may permit differentiation between normal underlying lung and pulmonary hypoplasia. We recommend serial scanning to detect early adverse effects of pleural effusion upon the fetus and to plan management changes. PMID- 3052984 TI - Clinical pharmacokinetics of enzyme inhibitors in antimicrobial chemotherapy. AB - The effectiveness of some antimicrobial agents can be enhanced by using them in combination; such combinations are termed synergistic. Where one compound potentiates the effect of a second drug they may be coformulated. Inhibition of the bacterial degradation of an active antimicrobial is the basis of clavulanate and sulbactam-potentiated penicillin combinations, and inhibition of degradative pathways in the host is the rationale behind imipenem/cilastatin therapy. Trimethoprim/sulphonamide combinations depend on the maintenance of an effective ratio for synergistic action. In order to achieve potentiation the coformulated drugs should have similar pharmacokinetics. Trimethoprim was originally matched with sulphamethoxazole, since these two drugs have similar elimination half lives, but the significantly poorer penetration of sulphonamides, their greater non-renal clearance, the emergence of resistance, and the adverse reactions attributable to them argue against the rationale that underlies their coformulation. Time-dependent inhibition of bacterial beta-lactamases by clavulanic acid and sulbactam has extended the use of penicillins which are highly susceptible to beta-lactamase inactivation. The beta-lactamase inhibitors must penetrate to the same extent as the penicillin used with them, and be present long enough to effect inhibition; thus, rapid penetration, similar or slower elimination and equivalent volume of distribution are necessary. These requirements are met for amoxycillin/clavulanic acid, ticarcillin/clavulanic acid and ampicillin/sulbactam combinations. Clavulanic acid is absorbed orally and is given with amoxycillin. However, since sulbactam is labile by this route, the combination of sulbactam with ampicillin to form the prodrug sultamicillin has been necessary to enable an oral form to be developed. Imipenem is metabolised by renal brush-border dehydropeptidases, and may cause proximal tubular necrosis. Cilastatin was designed to inhibit this metabolism, which it effectively does, thereby both potentiating the effect of imipenem and avoiding toxicity. Appropriate matching of the kinetics of coformulated drugs is intended to maximise potentiation and minimise the risk of emergent resistance. The kinetics of the above combinations are discussed in the light of these requirements and the effects of age and disease. PMID- 3052997 TI - Ultrasound diagnosis of adenomyomatosis of the gall-bladder: ultrasonic and pathological correlation. AB - Nine cases of adenomyomatosis of the gall-bladder are reported in which the ultrasound appearances are compared with those of oral cholecystography and the pathological findings. It is suggested that the ultrasound appearances are specific in the established disease. PMID- 3052998 TI - An unusual appearance of renal lymphoma. AB - The kidney is a relatively common site of extranodal involvement by lymphoma and various patterns have been documented (Richmond et al., 1962). We describe a patient in whom renal involvement by non-Hodgkin's lymphoma produced unusual appearances on ultrasound and computed tomography (CT). The possible causes and relevance of these findings are discussed. PMID- 3052999 TI - Digital subtraction angiography--a critical analysis. AB - The advantages and disadvantages claimed for intravenous and intra-arterial digital subtraction angiography (DSA) are critically examined. The parameters determining image quality and the factors degrading the image in intravenous DSA are discussed. It is argued that many of the patients specifically referred for intravenous digital subtraction studies are unsuitable for this examination either because poor image quality is to be expected and/or because the large contrast load inherent in this type of examination is undesirable. PMID- 3053000 TI - The value of CT in the diagnosis of recurrent carcinoma of the cervix. AB - The results of computed tomography (CT) and other imaging techniques performed on 70 patients who were investigated for suspected recurrent carcinoma of the cervix are reported. Recurrent disease was present in 39 patients. In 29, there was local recurrence with or without distant metastases and there was distant recurrence only in 10. Computed tomography correctly assessed the presence of local recurrent disease in 85% of patients. Six equivocal, two false positive and two false negative CT examinations in the assessment of local recurrence were due either to difficulty in differentiating recurrent disease from changes following radiotherapy, or to the failure of CT to detect small areas of local recurrent disease. Ultrasound and lymphangiography each detected recurrence in one patient which was missed by CT, but this was the most reliable technique for the detection of both local and distant recurrent disease. PMID- 3053001 TI - The role of ultrasound scanning in management after liver transplantation. AB - A total of 368 ultrasound scans in 72 patients following liver transplantation have been reviewed. Thirty-seven intra-abdominal fluid collections were identified. Twenty-four resolved spontaneously and 13 required aspiration under ultrasound control. Dilated bile ducts were seen in 22 patients, 11 of whom required further intervention. The main indications for intervention post transplantation were increasing or late onset (after 3 months) duct dilatation. Recurrent or persistent fluid collections particularly in the right subhepatic space also indicate further investigation. Twelve ultrasound-guided liver biopsies were performed to detect rejection or tumour recurrence. Eight cases of portal vein anastomotic narrowing and three abnormalities of the inferior vena cava were also detected. PMID- 3053002 TI - Assessment of gallbladder function by ultrasound: implications for dissolution therapy. AB - Successful treatment of gallstones by oral dissolving agents depends upon a functioning gallbladder. Thirty-one patients were assessed by oral cholecystography and ultrasound before and after a fatty meal. A correlation was demonstrated between radiological opacification and contraction on ultrasound. It is suggested that assessment of gallbladder function may be made by ultrasound methods alone, and an oral cholecystogram is not necessary prior to dissolution therapy. PMID- 3053003 TI - Ultrasound detection of oesophageal varices--comparison with endoscopy. AB - In a prospective study of 100 patients for the detection of oesophageal varices, real-time ultrasound was both sensitive (sensitivity 82%) and specific (specificity 91%) compared with upper gastrointestinal endoscopy. All patients with large and medium sized varices were correctly identified by ultrasound. Ultrasound should be considered as an alternative to endoscopy and radiology for detection of oesophageal varices. PMID- 3053004 TI - Multiple cholesterol emboli syndrome complicating angiographic techniques. AB - Three cases of widespread microembolisation of cholesterol crystals following angiography are described. The multiple cholesterol emboli syndrome has been well described as a spontaneous phenomenon and it is surprising that its occurrence during angiography appears rare. Only 19 cases could be found in the literature and these are reviewed. Dissemination of particulate cholesterol material produces irreversible organ ischaemia when a threshold 'dose' is reached. Males and patients with clinical evidence of widespread atherosclerosis are at increased risk. Prognosis is poor and available therapy unsatisfactory. PMID- 3053005 TI - Anterior extrathecal ependymoma with systemic metastases--a case report and review of the literature. AB - A case of anterior extrathecal ependymoma presenting with lung metastases is described. Computed tomography demonstrated the full extent of the mass which was not suspected on myelography. PMID- 3053007 TI - How to recognize a kickback and avoid unwitting entanglement in the Inspector General's net. PMID- 3053006 TI - Equipoise and the ethics of segmental liver transplantation. PMID- 3053008 TI - Radiologists speak out on RVS. PMID- 3053009 TI - Concurrent malaria and bacteraemia in a multiply transfused patient. PMID- 3053010 TI - Effect of 10 minutes heat treatment on HIV antibody testing from alternate testing sites. AB - One hundred and thirty-five sera were tested for the presence of antibody to human immunodeficiency virus (HIV) by enzyme immunoassay (EIA) with the Abbott Diagnostics HTLV-III test before and after heating at 56 degrees C for 10 min. Only one serum specimen was repeatedly reactive after heating when compared with the unheated portion (1/135). The specimen was a low level positive (ratio of test over cutoff = 1.6). Fifty-eight different sera, selected to yield a high percentage of positive results, had both their heated and unheated Western blot (WB) results agree completely with repeated EIA testing. An additional 4,244 sera heated at 56 degrees C for 10 min were tested. Of the 330 repeatable EIA positives, only 38 were not confirmed by WB. HIV infectivity in sera was reduced from 10(3.5) TCID50 to 10(1) TCID50 by the same heat treatment. We conclude that heating sera at 56 degrees C for 10 min significantly reduces HIV infectivity and does not significantly affect the results of HIV antibody testing. PMID- 3053011 TI - Comparison of the Abbott and Ortho enzyme immunoassays and cell culture for the detection of respiratory syncytial virus in nasopharyngeal specimens. AB - A comparison of the Abbott Laboratories and the Ortho Diagnostic Systems Respiratory Syncytial Virus (RSV) Enzyme Immunoassays (EIA) and HEp-2 cell culture for the detection of RSV in 81 nasopharyngeal (NP) specimens from pediatric patients with lower respiratory tract infection was carried out. The sensitivity and specificity of the Abbott test compared to confirmed infection was 92.3% and 100.0%, respectively. The sensitivity and specificity of the Ortho test was 87.5% and 80.3%, respectively. We found the Abbott EIA test to be sensitive, specific, rapid, and easy to perform. PMID- 3053012 TI - Phototherapy and photochemotherapy in dermatologic diseases. AB - Phototherapy and photochemotherapy have been shown to be effective treatments in a number of cutaneous disorders, but they are not without risk. PUVA has revolutionized the treatment of psoriasis and stimulated major improvements in the administration of UVB therapy. Successful treatment depends on careful patient selection and evaluation. Accurate dosimetry and close supervision are necessary to avoid complications. Regular and careful follow-up, both during and after therapy, can help to identify and treat long-term side effects such as non melanoma skin cancers. In order to achieve the necessary balance between benefits and risks with these valuable forms of therapy, patients should be referred to dermatologists with special training. PMID- 3053013 TI - Premenstrual symptoms: evaluation and treatment. AB - Critics who claim there is no therapy for PMS should now recognize that there are many ways to treat this condition. Appropriate therapy for PMS is more than a pill, more than changing attitudes, more than removing stress from one's life. Indeed, therapy for PMS is not always easy, for the problems that preceded the PMS may persist and the problems created by the PMS may be slow to heal. However, the good news is that most women with PMS will experience significant improvement in their sense of well-being and their quality of life with appropriate and multidisciplinary therapy. PMID- 3053014 TI - Anesthesiology update: the preanesthetic evaluation. AB - Ideal anesthetic management consists of the preanesthetic attainment of a state in which a diagnostic or therapeutic procedure or surgical operation can be performed with as little physiologic and psychologic trauma as possible, and the likelihood of a speedy and uneventful postoperative course is enhanced. Significant impairment of cardiovascular, respiratory, renal and other organ system functions is common in patients facing surgery. Thus, it is imperative that preoperative evaluation be complete. Signs of congestive heart failure, myocardial ischemia or infarction or both, hypertension, electrolyte abnormalities, and cardiac arrhythmias should be aggressively sought and treated before surgery. In addition, before the induction of anesthesia, information must be obtained about the specific associated pathologies (cardiac and otherwise), drugs being used to treat these conditions, and the adequacy of the current therapies. The preoperative evaluation does not begin with the anesthesiologist's visit, but with the evaluation and care given by the primary physician. A team approach is advisable in complex cases. Appropriate consultations, including that of the anesthesiologist, should be made in a timely fashion. Except in emergency situations, the temptation to send incompletely evaluated and treated patients to the operating room should be resisted. Good preoperative preparation will improve the chances for a successful outcome. PMID- 3053015 TI - Malignant hyperthermia. PMID- 3053016 TI - Update: treatment of rheumatoid disease. AB - The treatment of rheumatoid arthritis is, of necessity, highly individualized and thus invites the full application of the art of medicine. Patient education, physical therapy, NSAIDs, DMARDs, and surgical therapy all potentially play a role in the management of this syndrome. Careful and repeated assessment of signs and symptoms will identify patients who are likely to benefit from the wide array of helpful but imperfect drugs currently in use. The availability of increasingly effective therapy means that early referral and treatment are more important than ever in treatment of rheumatoid arthritis. PMID- 3053017 TI - Diagnosis and treatment of diabetic foot infections. PMID- 3053018 TI - High-frequency hearing of dental personnel. AB - 234 dentists and dental nurses were examined with a normal and a high-frequency audiometer in high standard clinical conditions. Their ordinary and high frequency hearing as compared with the controls showed no significant differences. Thus, exposure to high-frequency noise from high-speed drills and other modern dental instrumentation does not appear to be harmful to one's hearing and does not necessitate audiologic screening procedures for dental personnel. PMID- 3053019 TI - Prevalence study of oral leukoplakia in a selected population of 1000 patients from The Netherlands. AB - A survey of a special population, the outpatients of a Department of Oral Surgery of a Teaching Hospital in Amsterdam, revealed a comparatively low prevalence rate for oral leukoplakia of 14/1000 patients (1.4%). No difference was found in the age distribution compared with similar investigations elsewhere in the world. However, in contrast with previous findings, which show a significant difference between prevalences of leukoplakia for men and women, leukoplakia was equally distributed between both sexes. PMID- 3053020 TI - The chronically mentally ill case management--more than a response to a dysfunctional system. AB - The service systems which assist the long-term mentally ill to function in the community have been routinely described as fragmented and uncoordinated. The development and implementation of case management has been seen as one response to this dysfunctional system. This article examines case management from the perspective that case management is a needed function no matter how coordinated and integrated the system. From this perspective, case management is driven by the clients' goals and not the systems' goals. Case management is viewed as a process by which the person with severe psychiatric disability is supported in negotiating for the various services that they want and need. Four unique activities are identified as performed by the case manager: Connecting with Clients, Planning for Services, Linking Clients with Services, and Advocating for Service Improvements. Case management must be seen as a uniquely human response to the client's specific service needs and overall goals. For persons with long term psychiatric disabilities, case management brings to life the human dimension of the human service system. PMID- 3053021 TI - [An immunoenzyme test used for the detection of antibodies to rinderpest: the advantage of using a purified virus cultivated in a cell line]. AB - Two types of in vitro assays (enzyme immunoassay and sero-neutralization test) were compared for their ability to detect antibodies to rinderpest virus in field sera from West African bovines. Purified virus grown on Vero cells (Ag/Vero) or bovine kidney cells (Ag/BK), were tested as antigen in enzyme-linked immunoassays (EIA). The results of the comparative evaluation of the two antigens by EIA, prove that Ag/Vero did enhance the sensitivity and the specificity of the test in comparison to Ag/BK and offers a 94% correspondence with seroneutralization. PMID- 3053022 TI - Antibiotic resistance of Escherichia coli strains isolated from chickens with colisepticaemia in Morocco. AB - Sixty-two strains of Escherichia coli were isolated in 58 farms from broiler chickens showing respiratory signs and lesions characteristic of avian colibacillosis. Serological examination of these strains showed that the types 078, 01 and 02 (for the somatic antigen) and K1 (for the capsular antigen) were the most frequently found. Newcastle disease virus was also isolated in two cases. All the strains of E. coli isolated were sensitive to colistin, flumequine and gentamicin. A few strains were resistant to neomycin, nalidixic acid and trimethoprim. The frequency of strains resistant to nitrofurans, sulfonamides, chloramphenicol, spectinomycin and ampicillin was intermediate. Most strains were resistant to tetracycline. Multiple resistance was common. PMID- 3053023 TI - Immunopathogenesis of canine angistrongylosis: pulmonary effects of infection. AB - Deposits of immunoglobulins, complement, and fibrinogen were localized in the lungs of Angiostrongylus vasorum-infected dogs. In acutely infected dogs, significant deposits of IgA, IgG and IgM, complement, and fibrinogen were seen. The predominant immunoglobulin noted in infections of longer duration was IgG. The continued presence of fibrinogen as the predominant protein deposited suggested a continuation of intravascular coagulation. This was borne out by demonstrating luminal deposits of fibrinogen in many blood vessels. Histopathologic examination of the large revealed lesions typical of angiostrongylosis, as well as eggs of the worm as early as 35 days postinfection. This is the earliest report of egg production by A. vasorum on record. PMID- 3053024 TI - Norm-of-reaction: definition and misinterpretation of animal research. AB - The development of a phenotype is due to an interaction of the genotype with the environment. Two terms have been used to describe the outcome of this interaction, the norm-of-reaction and the reaction range. The first represents the theoretically limitless distribution of the phenotypes that may be expressed by a given genotype. The reaction range implies an upper and lower limit for phenotype expression possible from a given genotype. A critical distinction between the reaction range and the norm-of-reaction is that the norm-of-reaction is a statement of the conceivable interactions found but does not imply any predictability other than that within the conditions previously tested experimentally, that is, the tails of a normal distribution are infinitely variable, whereas the concept of reaction range implies a limitation inherent in the genotype, that is, a finite range. Empirical support for the reaction-range concept is questionable. Animal studies cited in support of the reaction range have been inappropriately and incorrectly interpreted. PMID- 3053025 TI - Enhancing effect of autologous human erythrocytes on generation of C3 cleavage products beyond iC3b. AB - The in vitro formation of C3d and C3c in fresh normal human serum (NHS) after addition of five different activators of the complement (C) system was studied. Following C-activation in NHS (n = 53) by Sephadex G-200 beads, the conversion of C3 was found to proceed to iC3b with a variable but restricted generation of C3d. Similar results were obtained by use of heat-aggregated IgG, Escherichia coli, zymosan, and cobra venom factor. However, comparing the C3d concentration following activation in the presence and absence of autologous red blood cells (RBC) at 37 degrees C the generated C3d was found to be 2- to 3-fold higher in the presence of RBC after 30, 60, and 210 min. Preincubation of RBC with polyclonal anti-CR1 antibodies resulted in a dose-dependent reduction of the amount of C3d generated. C-activation induced by Sephadex G-200 beads, in the absence of RBC, generated iC3b without a significant production of C3d. After removal of the activator beads, addition of RBC resulted in a decrease of iC3b and a clear increase in the C3c and C3d concentration within 3 h. Western blotting analysis of the C3d produced in the presence of RBC showed that the molecular weight (36 kilodaltons) was similar to that of C3d formed in vivo. PMID- 3053026 TI - Self-service computerized bibliographic retrieval: a comparison of Colleague and PaperChase, programs that search the MEDLINE data base. AB - Colleague and PaperChase are the two most widely used computer systems designed specifically for clinicians and scientists who wish to search the National Library of Medicine's MEDLINE data base of references to the biomedical literature. The present study compares the performance of these two systems. Two matched groups of second-year medical students each received 3 hr of instruction, one group in Colleague, the other in PaperChase. Each student then attempted 10 test searches. The next day the groups were reversed, and each student attempted 5 additional searches. During 3 1/2 hr allocated for searching, users of Colleague attempted 64 test searches and retrieved 326 target references; users of PaperChase attempted 78 searches and retrieved 496. Users of Colleague took a mean of 2.2 min and spent a mean of $1.20 to find each target reference; users of PaperChase took 1.6 min and spent $0.92. We conclude that after limited training, medical students find more references faster and at lower cost with PaperChase than with Colleague. PMID- 3053027 TI - Depressive symptoms in schizophrenia: comprehensive differential diagnosis. AB - Depression is a common complication of schizophrenia and is associated with increased morbidity and mortality. Contrary to traditional clinical wisdom, depressive symptoms occur during all phases of schizophrenia and are not restricted to the postpsychotic period. In this review, the authors summarize current empirical research and offer a practical approach to the identification of depressive subtypes in schizophrenia. The following subtypes are considered: (1) depressive symptoms occurring secondary to organic factors (caused by medications, substance abuse, or underlying medical problems); (2) nonorganic depressive symptoms occurring with acute psychotic symptoms (intrinsic to the acute psychotic episode or schizoaffective disorder); and (3) nonorganic depressive symptoms occurring without acute psychotic symptoms (prodromal symptoms, negative symptoms, acute dysphoria, secondary depressive syndrome, or chronic demoralization). The authors discuss each of these entities and offer guidelines for diagnosis. PMID- 3053028 TI - Contact sensitivity in otitis externa. PMID- 3053030 TI - Renal oncocytoma: a benign tumor often misdiagnosed as malignant. Case reports and a diagnostic protocol. PMID- 3053029 TI - Alterations of epidermal lectin binding sites in acute contact dermatitis. AB - We investigated alterations of epidermal lectin binding sites, as well as of pemphigus and bullous pemphigoid antigens, in 28 human patch test reactions, both allergic (nickel, formaldehyde, N,N'-1,3-dimethylbutyl-N'-phenylenediamine) and irritant (sodium lauryl sulfate). The epidermal reactivity to a panel of lectins and human antisera to pemphigus vulgaris and bullous pemphigoid antigens was compared with samples obtained from normal skin and from skin under tape occlusion. We observed selective perturbations of lectin and antibody binding in acute contact dermatitis, whether allergic or irritant. The main findings were a loss of terminal sialic acids and longer bi- and triantennary mannosyl residues as well as a loss of pemphigus vulgaris antigen. The only difference between allergic and irritant patch test reactions was in topography of loss of WGA binding sites: in the former, it was most pronounced in the lower and middle epidermis, whereas in the latter it was seen in the uppermost subcorneal layers. Our findings support a common pathway of cell membrane alterations of keratinocytes in acute contact dermatitis. PMID- 3053031 TI - Consensus conference: Perioperative red cell transfusion. National Institutes of Health. PMID- 3053032 TI - Lost guide wires: a case report showing a complication of central vein cannulation. PMID- 3053033 TI - The parathyroid gland and aluminum overload: an overview. PMID- 3053034 TI - Parathyroid hormone secretion: perturbations in chronic renal failure. PMID- 3053035 TI - Vitamin D and the kidney: a short review. PMID- 3053037 TI - Does lead play a role in the development of renal insufficiency? PMID- 3053036 TI - Disturbed growth in uremia: are hormonal factors responsible? PMID- 3053038 TI - Factors modifying the secretion and action of parathyroid hormone in renal failure. PMID- 3053039 TI - Calcium carbonate and magnesium hydroxide in the prevention of renal osteodystrophy or the demise of aluminum toxicity in uremia. Analysis of 5 years experience. PMID- 3053040 TI - Mechanisms of metal-induced renal cell injury: roles of high-affinity metal binding proteins. PMID- 3053041 TI - The statistical analysis of graft patency data in a clinical trial of antiplatelet agents following coronary artery bypass grafting. AB - Because most coronary artery bypass patients receive more than one graft at surgery, it is most important to determine whether statistical analysis of graft patency should be performed on the premise that the multiple grafts within patients are dependent or independent experimental units. Veterans Administration Cooperative Study No. 207 was a multicenter clinical trial comparing four different antiplatelet regimens to placebo in the prevention of graft occlusion following coronary artery bypass grafting. Using the results from the 1-week postoperative angiograms from the Veterans Administration Cooperative Study No. 207, in which there were 3.2 distal anastomoses per patient, we have tested the hypothesis that grafts within patients tend to act dependently with respect to patency or occlusion by comparing the graft patency data to a binomial distribution (i.e., that distribution that would have been manifest if grafts were independent). Because the graft patency results in Study No. 207 significantly deviated from the binomial distribution (p = 0.0003), a more appropriate analysis for graft patency data was applied using a ratio estimate as applied to cluster sampling. The statistical methods used in 11 previous clinical trials of antithrombotic therapy after coronary artery bypass grafting were examined. Only one of the previous studies used such an analysis, and three additional reports attempted to correct for dependency of grafts within patients in their analyses using other statistical methods. In seven of the studies the investigators did not address the potential problem of a dependent relationship between multiple grafts within patients. We conclude that grafts within patients act as dependent experimental units and that the ratio estimate as applied to cluster sampling may be appropriately applied to these data. PMID- 3053042 TI - A one-sided interim analysis with binary outcomes. AB - Wieand and Therneau (Controlled Clin Trials 8:20-28, 1987) proposed a technique for conducting a clinical trial to test the equality of response rates for two treatments using a one-sided test with an option of terminating the trial at the single interim analysis if it appears that the test treatment will offer no improvement over the control treatment. Chi, Bristol, and Castellana (Stat Med 5:387-392, 1986) considered a clinical trial with the same option when the treatments are compared with respect to the means of variables with normal distributions with a common known variance. Here a decision rule similar to the one proposed in the latter is employed for the problem examined in the former. This decision rule, a generalization of the one proposed by Wieand and Therneau, consists of a one-sided test at the final analysis with a one-sided test at the interim analysis, which is performed in the direction opposite to the one at the final analysis. It is shown that the proposed generalization may result in desirable properties regarding the probability of stopping the trial at the interim analysis in some situations. PMID- 3053043 TI - Sensory feedback control of upper- and lower-extremity motor prostheses. AB - The topic is introduced with a conceptual definition of sensory feedback control applied to prosthetic systems. An overview of sensory physiology is then provided. Tactile sensation and audition are specifically discussed. Next, the artificial sensory systems are evaluated. This is done with respect to sensory (input) transducers, signal encoding/decoding, and sensing output transducers. Specific prosthetic systems are then reviewed. First, upper-extremity prostheses are reviewed with respect to artificial arm/hand systems as well as neuroprostheses. Next, lower-extremity prostheses are reviewed, and include both artificial leg and foot systems as well as neuroprostheses. Finally, conclusions are drawn regarding effective state-of-the-art sensory feedback control used with orthotic and prosthetic systems. PMID- 3053045 TI - Magnetic resonance imaging of musculoskeletal tumors. AB - Magnetic resonance (MR) has been shown to provide superior soft tissue contrast as well as superior anatomical definition when compared with other diagnostic modalities in the evaluation and staging of many bone and soft tissue tumors. MR signal patterns with various parameter weighting can provide information that aids in the tissue characterization of the lesion. Technical factors as well as the clinical role of MR in the evaluation of musculoskeletal tumors are discussed. PMID- 3053044 TI - Radiologic contribution to the management of patients undergoing extracorporeal shock wave lithotripsy. AB - Since its first clinical application in 1980, the use of extracorporeal shock wave lithotripsy (ESWL) has dramatically changed the treatment of urinary tract stone disease. More than 80% of patients with urolithiasis will undergo ESWL as a first line therapy. Detection of stones, appropriateness of ESWL treatment, successful stone therapy, termination of the ESWL procedure, and evaluation of treatment complications are all dependent on radiologic input. The application of adjuvant interventional radiologic percutaneous techniques will extend the use of ESWL to an additional 15% of patients and reduce the need for surgical intervention. Additionally, percutaneous techniques can be used to improve success rates in problematic cases and to treat complications related to the ESWL procedure. This article discusses the pre- and postprocedural radiologic evaluation of patients undergoing ESWL. Adjuvant interventional radiologic techniques useful in the management of these patients are also discussed. PMID- 3053046 TI - Radiological detection and diagnosis of pouch of Douglas lesions. AB - The pouch of Douglas (cul-de-sac) represents the caudal extension of the peritoneal cavity. It is the rectovaginal pouch in the female and the rectovesical pouch in the male. The cul-de-sac is in a dependent position when either upright or supine; it is, therefore, a frequent location for seeded lesions. Abnormalities in the cul-de-sac include metastases, abscesses, and endometriosis. These lesions may be detected by multiple modalities, including barium enema, computed tomography, and ultrasound. Examples of numerous cul-de sac lesions are presented as they appear with different imaging modalities. PMID- 3053047 TI - Picture archiving and communication systems (PACS) for radiological images: state of the art. AB - Implementation of a Picture Archiving and Communication System (PACS) is a system integration task and requires the knowledge of multidisciplinary fields. This paper reviews current PACS development with emphasis on radiological images. The following topics are covered: methods of image acquisition, image compression, storage, display, communication, and image database structure. Methods of implementation of PACS in a clinical environment as well as current operational PACS in hospitals are reviewed. A survey of private industry participating in PACS research and development are also given. PMID- 3053048 TI - Bacterial alternative nitrogen fixation systems. AB - The introduction briefly reviews some of the salient features of the well characterized conventional molybdo-enzyme system for N2 fixation. This is followed by a brief account of the discovery of an alternative N2 fixation system that does not require molybdenum in the N2-fixing bacterum Azotobacter vinelandii. The next section cites observations from the early literature on N2 fixation suggesting may not always require molybdenum. Next, recent evidence for an alternative N2 fixation system in A. vinelandii is discussed. A brief description of our discovery of an alternative nitrogenase which is not a molybdenum or vanadium enzyme is presented, followed by a summary of recent papers describing an alternative vanadium-containing nitrogenase. Available information on the genetics and regulation of alternative N2 fixation systems is discussed. Finally, the possible/probable presence of alternative N2 fixation systems in bacteria other than Azotobacter species is covered. PMID- 3053049 TI - The bioenergetics of alkalophilic bacilli. AB - A summary, cum speculation, of the major bioenergetic characteristics of alkalophilic bacilli is presented in Figure 5. Further progress will depend heavily on the purification and characterization of the relevant proteins that catalyze the ion fluxes and on the development of much more potent genetic approaches to the outstanding issues of this interesting group of bacteria. PMID- 3053050 TI - The ecology and pathogenicity of urease-producing bacteria in the urinary tract. AB - Urease activity is a physiological function of many bacteria that enables these organisms to utilize urea as a source of nitrogen. The association of ureolytic bacteria with human or animal hosts varies widely from a commensal relationship as demonstrated with skin microflora, a symbiotic relationship in the gastrointestinal tract, to a pathogenic relationship in the urinary tract. Since similar or identical species of bacteria such as Staphylococcus aureus are found in all three environments, the effect of urease activity on the host must be solely a function of the environment of these organisms. In this review, the importance of urease to bacteria is discussed, identifying the gastrointestinal tract as a major reservoir of ureolytic bacteria and investigating the urinary tract environment and the infectious struvite stone production that often accompanies urease-producing bacteria there. Finally, an infection model is presented which explains the development and growth of these urinary calculi and their remarkable persistence in spite of modern urological treatments. PMID- 3053051 TI - Pleurotus mushrooms. Part II. Chemical composition, nutritional value, post harvest physiology, preservation, and role as human food. AB - The fruit bodies of Pleurotus species as a class of "Edible Fungal Foods" have been discovered to have definite nutritive and medicinal values. They are a good source of nonstarchy carbohydrates, dietary fiber (that can help in reducing the plasma cholesterol), most of the essential amino acids, minerals and vitamins of B group, and folic acid (necessary to counteract pernicious anaemia) in particular. Considering the essential amino acid index, biological value, in vitro digestibility, nutritional index, and protein score, Pleurotus species fall between high grade vegetables and low grade meats. Fractions of water-soluble polysaccharides are reported to possess antitumor activity. The physiological processes such as changes in water content, respiratory rate, texture, color, and activities of enzymes like proteases and polyphenol oxidases during the after harvest life are delineated. The problems and prospects of processing the fruit bodies by various methods are discussed. Potentialities for production and consumption of the fruit bodies in different parts of the world are brought out. PMID- 3053053 TI - Cardiac transplantation. PMID- 3053052 TI - HLA DR and DQ distribution in normal human ocular structures. AB - HLA DR and DQ distribution was investigated in normal human ocular tissues, together with class I antigens and immunocompetent cell subsets, by immunofluorescence and immunoperoxidase procedures. In the anterior segment, our findings, consistent with those of previous reports, showed the wide distribution of class I antigens, specially in the corneal epithelium, while class II antigens were restricted to very rare cells scattered in the conjunctiva, the peripheral cornea and the stroma of the ciliary processes. Some non pigmented epithelial cells of the ciliary processes were HLA DR and DQ positive. In the posterior segment, class I antigens were abundantly represented in the choroid and the retinal layers. Few HLA DR and DQ positive cells were seen in the choroid, similar to those found in the anterior segment. Normal RPE did not react with any monoclonal antibody, but numerous cells located in the retina were strongly HLA DR and DQ positive, all around the blood vessels, and not at the sites of endothelial cells. The characterization of those cells, which could be hypothetized as pericytes needs further studies but suggests close relationships between neuroretina and the immune system. This study may provide insight in the implication of the immune system in many poorly understood ocular diseases. PMID- 3053054 TI - Immunocytogenetics: localization of transcriptionally active rRNA genes in nucleoli and nucleolus organizer regions by use of human autoantibodies to RNA polymerase I. AB - Cytological staining with silver nitrate (AgNO3) has proved useful for the localization of nucleoli in interphase nuclei, as well as of nucleolus organizer regions (NORs) in metaphase chromosomes. The affinity of interphase nucleoli and chromosomal NORs to silver is a direct measure of the ongoing transcriptional activity of the rRNA genes or their activity during the preceding interphase, respectively. Correspondingly, human autoantibodies directed against chromatin associated RNA polymerase I (RPI) should also be of value in the investigation of transcribed rRNA genes. Indirect immunofluorescence using the anti-RPI antibody as a probe has been employed successfully to visualize the chromosomal distribution of NORs in various mammalian species, as well as in human tumor cells. Immunofluorescence staining even permits the identification of heteromorphisms and small aberrations of the chromosomal NORs. The fluorescent intensity of interphase nucleoli is correlated with the different stages of nucleolar activation. In male gametogenesis, RPI-positive granules are present during meiotic prophase up to pachytene, as well as during the early and middle spermatid stages. PMID- 3053056 TI - Oncogene expression detected by in situ hybridization in human primary lung cancer. AB - By means of in situ hybridization using biotinylated oncogene probes and the immunohistochemical reaction of avidin-biotin complex-alkaline phosphatase with substrate, we investigated expression of c-myc oncogene in the formalin-fixed, paraffin-embedded tissue sections from seven patients with squamous cell carcinoma (six cases) and small cell carcinoma (one case) of primary lung origin. The expression of c-myc oncogene was greatly enhanced in all cases studied, with individual and cell-to-cell variation. In contrast, all of the specimens incubated with deoxyribonuclease after the standard pretreatment with ribonuclease T1 were negative for the expression of c-myc oncogene. The in situ hybridization permits estimation of a heterogeneous amplification of c-myc oncogene that may be related to secondary alterations occurring during the progression of the malignant lung tumors. PMID- 3053055 TI - Neuropsychologic symptoms in obstructive sleep apnea improve after treatment with nasal continuous positive airway pressure. AB - To describe the affective changes associated with sleep restoration we assessed psychologic symptoms using the Profile of Mood States questionnaire before and two months after treatment with nasal continuous positive airway pressure (NCPAP) in seven men with obstructive sleep apnea (OSA). The results were compared with those of a control group of patients with OSA who did not receive NCPAP. Two of six mood factors, depression and fatigue, improved significantly following treatment with NCPAP. Total Mood Disturbance (TMD) score was used to assess global mood differences. The mean TMD score for the patients before treatment was 1.7 and during treatment decreased to -7.6 (p less than 0.05). This mean decrease of 9.3 in the TMD score implies generalized improvement in mood. These findings support the opinion that sleep fragmentation and abnormalities of respiration during sleep are at least partially responsible for affective changes seen in sleep apnea. These psychologic disturbances improve after treatment with NCPAP. PMID- 3053057 TI - Leukocyte migration inhibition in amiodarone-associated pneumonitis. AB - Amiodarone-associated pneumonitis is now a well-known clinical entity, but the mechanism for the induction of the pulmonary disease is ill defined. In four patients with this disorder, evidence was obtained for elaboration of a lymphokine, leukocyte inhibitory factor (LIF), by peripheral blood lymphocytes after incubation with amiodarone in the direct leukocyte migration inhibition test. Control lymphocytes from normal subjects, as well as from patients receiving amiodarone but without pneumonitis, failed to elaborate LIF in the presence of the drug in this test. This production of LIF suggests that pneumonitis associated with amiodarone therapy is also associated with a specific cellular immune response to the drug. PMID- 3053058 TI - Aneurysms of the pulmonary arteries. PMID- 3053059 TI - New drugs for treating lung infection. PMID- 3053060 TI - Pulmonary edema due to upper airway obstruction in adults. AB - A report of pulmonary edema following acute upper airway obstruction in an adult is presented, and the literature involving 25 additional cases is reviewed. This form of pulmonary edema appears to be related to markedly negative intrathoracic pressure due to forced inspiration against a closed upper airway resulting in transudation of fluid from pulmonary capillaries to the interstitium. Postanesthetic laryngospasm is the most common cause of pulmonary edema in adults (11/26 cases). The edema usually clears rapidly with supportive care. Aggressive diagnostic and therapeutic interventions may be avoided if the syndrome is recognized. Maintenance of oxygenation and a patent airway are the mainstays of treatment. PMID- 3053061 TI - Horner's syndrome secondary to hydromediastinum. A complication of extravascular migration of a central venous catheter. AB - We report the findings in a patient who developed Horner's syndrome as the first manifestation of mediastinal migration of a central venous catheter that resulted in hydromediastinum and hydrothorax. The pathogenesis of this complication of central venous catheterization is discussed. PMID- 3053062 TI - The impact of pediatric asthma education on morbidity. Assessing the evidence. AB - We reviewed the literature evaluating pediatric asthma education interventions to assess their impact on morbidity (school absences and health care utilization). Thirteen studies were analyzed, most of which reported favorable outcomes. Of the ten studies reporting on school absences, only seven used tests of statistical significance when reporting on postintervention reductions, and of those, only two found a significant decrease in absenteeism. Similarly, among the studies reporting on utilization, not all used tests of statistical significance when reporting on postintervention decreases in physician visits, ER visits, and hospitalization. Only four of the ten studies used adequate sample sizes to detect a 20 percent reduction in school absences, and stratification of the sample by severity of asthma suggests that some programs do reduce health care utilization among those children with more severe disease. We conclude that the effectiveness of asthma educational programs on reducing school absences and health care utilization may be small. These programs are best directed toward children with moderate or severe disease. Finally, it is important for pediatricians, children with asthma, and their families to have realistic expectations about what these programs may accomplish. PMID- 3053064 TI - Studies on antifungal agents. 17. Effects of the C-5-substitution on the in vitro activity of novel 3,5-substituted isoxazolidines. AB - A number of novel 5-(substituted thiomethyl)-3-phenyl-3-(1H-imidazol-1-ylmethyl) 2- methylisoxazolidine derivatives were prepared and evaluated in vitro for antifungal activity in solid agar cultures against Trichophyton rubrum, Aspergillus fumigatus and Candida albicans. The effect of the 5-(substituted thiomethyl) group on the degree and scope of activity was studied and compared to that of the corresponding 5-phenoxymethyl and 5-[(substituted phenyl)thio(or amino)methyl] derivatives. PMID- 3053063 TI - Effect of acute changes in heart rate on Doppler pulmonary artery acceleration time in a porcine model. AB - Doppler measurements of pulmonary artery (PA) acceleration time (AT) have been used clinically to estimate PA pressure. However, these studies have been performed primarily in patients without tachycardia. To determine the effect of acute changes in heart rate on PA AT, atrial pacing studies were performed in seven closed-chest pigs. Pulsed Doppler PA flow velocity recordings were obtained from a parasternal position at pacing rates from 100 to 140 beats/min. PA pressure remained constant (mean +/- SD = 14 +/- 5 mm Hg) over the entire range of paced rates. When PA Doppler measurements were compared at heart rates of 100 and 140 beats/min, there were decreases at the higher heart rate in both acceleration time (110 +/- 12 vs 83 +/- 11 ms, p less than 0.01) and ejection time (ET) (315 +/- 23 vs 237 +/- 21 ms, p less than 0.01). In contrast, there was no change in either PA peak flow velocity (69 +/- 15 vs 62 +/- 18 cm/s) or the ratio of AT/ET (0.35 +/- .02 vs. .36 +/- .03). Consequently, when estimating PA pressure in states of tachycardia, the PA AT/ET ratio may be a more useful measurement than PA acceleration time. PMID- 3053065 TI - In vitro and ex vivo enhancement of nonspecific phagocytosis by cefodizime. AB - The in vitro and ex vivo effects of cefodizime on some functional activities of both human neutrophils and monocytes were studied. In vitro experiments were performed with antibiotic concentrations ranging from 1 to 200 micrograms/ml. For the ex vivo study, 7 adult healthy controls were treated intravenously with 4 g/day of cefodizime for 6 days. We found that the drug modulated phagocytosis frequency and index when nonopsonized zymosan and heat-killed Candida albicans were used as phagocytic challenge both in vitro (from 25 micrograms/ml) and ex vivo 12 h after the last administration of cefodizime. No effect on the other phagocyte functional parameters was shown. The in vitro enhancement of nonspecific phagocytosis was demonstrated both in the presence of cefodizime and when phagocytes and particles were separately incubated with the drug. PMID- 3053067 TI - [Indirect immunofluorescent antibody test in the diagnosis of hydatid disease]. PMID- 3053066 TI - [Surgical and echographic comparison in Morton's neuroma]. PMID- 3053068 TI - [Determination of antinuclear autoantibodies in progressive systemic sclerosis and its clinical significance]. PMID- 3053070 TI - Interval-coded texture features for artifact rejection in automated cervical cytology. AB - In order to improve the separation between abnormal cells and noncellular artifacts in the CERVIFIP automated cervical cytology prescreening system, 22 different object texture features were investigated. The features were all statistical parameters of the pixel density histograms or one-dimensional filtered values of central and border regions of the object images. The features were calculated for 231 images (100 cells and 131 artifacts) detected as Suspect Cells by the current CERVIFIP and were then tested in hierarchical and linear discriminant classifiers. After selecting the two best features for use in a hierarchical classifier, 83% correct classification was achieved. One of these features was specifically designed to remove poorly focused objects. With maximum likelihood discrimination using all 22 features, an overall correct classification rate of 90% was obtained. PMID- 3053069 TI - [HLA and nonhemolytic transfusion reactions]. PMID- 3053071 TI - A new technique for treatment of pilonidal sinus. AB - Twenty-seven patients with chronic pilonidal sinus were treated by a new adopted technique. It was found to be simple, time-saving and to minimize the postoperative morbidity and hospital stay. Results of this new technique were compared with those of other excisional methods in the literature and were found to be superior to them with a shorter hospital stay. The new technique also preserves the internatal cleft, which is valuable in restoring the normal configuration of the breech. Six-year follow-up revealed no recurrences. PMID- 3053072 TI - Lymphangioma of the large intestine. Report of a case. AB - Lymphangiomas of the large intestine have been reported more frequently since the development and widespread use of the colonic fiberscope. It is possible to endoscopically diagnose lymphangiomas because they are lustrous and smooth on the surface, pliable on compression, and half of them have a stalk or a waist at the base. Lymphangiomas of the colon and rectum under 20 mm in diameter can be safely resected by electrocautery, thereby avoiding unnecessary operation. PMID- 3053073 TI - Relative roles of acid and mucosal compression in ulcerogenesis in indomethacin insulin-treated rat. AB - We have proposed that gastric peristaltic activity is primarily responsible for ulcerogenesis in the phenylbutazone-treated rat and that acid plays only a synergistic role. This study examines the effect of graded doses of the H2 blocker cimetidine on acid secretion and ulcerogenesis occurring during insulin induced peristalsis in the indomethacin (Indo) -pretreated rat. The second part of the study utilizes graded gastric distension with exogenous acid to examine the role of the forceful apposition of the mucosal folds during peristalsis in lesion genesis. It is demonstrated that the inhibition of acid secretion by cimetidine reduces but does not prevent ulceration. Gastric inflation with acid obliterates mucosal folding, prevents mucosal apposition during peristalsis, and abolishes ulcerogenesis. It is concluded that mucosal compression is the primary cause of the linear lesions along the base of the mucosal folds but that acid is necessary to extend the lesions once initiated. PMID- 3053075 TI - Colonic xanthomatosis. Relationship to disordered motility and review of the literature. AB - We report two additional cases of colonic xanthomatosis associated with persistent rectal symptoms. Disordered colonic motility in the areas of lipid infiltration was documented in one patient. We conclude these lesions may have a pathophysiologic role in the alteration of intestinal motility which appears to be the cause of our patients' symptoms. PMID- 3053074 TI - Slow moving proteinase in gastric cancer and its relationship to pepsinogens I and II. An immunohistochemical study. AB - Slow-moving proteinase (SMP), pepsinogen I (PG I), and pepsinogen II (PG II) are aspartic proteinases normally found in gastric mucosa. Because of differences in their cellular origins, normal gastric epithelial cells can be phenotyped by immunohistochemical staining with a panel of antisera to each proteinase. In this study, we determined aspartic proteinase phenotypes of malignant cells in 74 cases of gastric cancer by immunohistochemical staining with rabbit antiserum to human SMP, PG I, and PG II. Of the 74 cancers, 20 were histologically of the diffuse type and 54 were of the intestinal (or mixed) type. Intestinal metaplasia was characterized by the presence of SMP (but not PG I or PG II) in absorptive epithelial cells. SMP was found in 40 (54%) of the cancers (vs 31% for PG II and only 5% for PG I) and in both the intestinal and diffuse types. Of the 40 SMP positive cancers, 14 also expressed PG II. In the latter tumors, staining of adjacent sections revealed that some malignant cells expressed only SMP or only PG II, whereas others expressed both proteinases. Overall, 49 (66%) of the cancers contained cells with proteinase phenotypes characteristic of cells in nonmalignant gastric mucosa and 23% contained cells with proteinase phenotypes characteristic of more than one cell type. The results indicate that both intestinal- and diffuse-type gastric cancers often differentiate to cell types that produce aspartic proteinases and that about one fourth contain a heterogeneous population of malignant cells. PMID- 3053076 TI - Diseases of the adrenal cortex. AB - The adrenal cortex is functionally a three-dimensional gland that secretes glucocorticoids, mineralocorticoids, and sex steroids. Of these three classes of steroids only the gluco- and mineralocorticoid hormones are necessary to sustain life. The availability of sensitive and specific radioimmunoassays has permitted accurate measurement of practically every steroid hormone secreted by the adrenal cortex. As in other endocrinopathies, suppression studies are employed when hyperfunction is suspected, while provocative tests are used to detect hypofunction. These dynamic studies enable the clinician to evaluate the functional status of the adrenal cortex. The anatomic configuration of the adrenal cortices is delineated by high-resolution computed tomography (and magnetic resonance imaging), obviating the need for invasive procedures such as venography or arteriography. The disorders of the adrenal cortex can be viewed from the dual perspectives of hyperfunction and hypofunction. Clinical expressions of hyperfunctional adrenocortical syndromes include Cushing's syndrome, primary hyperaldosteronism, and the adrenogenital syndrome. The expressions of hypofunctional syndromes include Addison's disease and selective hypoaldosteronism. The diagnosis and treatment of these disorders are outlined in this issue. PMID- 3053077 TI - Malignant melanoma of the skin. PMID- 3053078 TI - Treatment of carbon monoxide poisoning. PMID- 3053079 TI - How long should antidepressive treatment continue? PMID- 3053080 TI - Contraindications to childhood immunisation. PMID- 3053081 TI - Maxepa--Eskimos, fish oils and ischaemic heart disease. PMID- 3053082 TI - [Effects of intensified insulin therapy on fat metabolism in type 1 diabetes mellitus]. AB - The effect on fat metabolism of an intensified insulin treatment regimen was tested in 23 type I diabetics and compared with that of conventional insulin treatment. Definite improvement in blood glucose levels was found after a one year intensified treatment schedule (HbA1 of 8.1% under intensified treatment, compared with 10.6% under conventional treatment). The intensified treatment schedule also resulted in a significant reduction in plasma triglycerides from an average of 114 mg/dl to 94 mg/dl (p less than 0.05), at the same time the plasma HDL-cholesterol levels increased. Plasma concentrations of total cholesterol and LDL-cholesterol remained unchanged. Thus intensified insulin treatment was shown to improve glucose metabolism, lower HDL-cholesterol and in this way improve the risk profile for macroangiopathy. PMID- 3053083 TI - [Diagnostic problems in neurosyphilis]. AB - In two patients admitted to hospital-one with signs of cerebral infarction, the other with headaches, vertigo and paraesthesias-the TPHA test was "reactive", while the 19S(IgM)-FTA-ABS test was not. There was no cerebrospinal fluid (CSF) pleocytosis. Further CSF analyses and serological tests for syphilis (including CSF protein profile, demonstration of oligoclonal IgG, quantitative determination of Treponema-specific antibodies in serum and CSF) confirmed the diagnosis of neurosyphilis requiring treatment. In both patients the biologically false negative 19S(IgM)-FTA-ABS test at first became transiently reactive after treatment. This unusual finding was probably due to antigen, liberated by treatment, again stimulating previously blocked IgM antibody synthesis. The listed additional tests should be performed in all patients with a reactive TPHA test and neurological or psychiatric signs and symptoms. PMID- 3053085 TI - [Diagnosis of deep venous thrombosis of the legs using real-time ultrasonography]. PMID- 3053084 TI - [Calcium antagonists in the cytostatic therapy of malignant tumors]. PMID- 3053086 TI - [Three-dimensional imaging in ultrasonic diagnosis. Initial results]. AB - A new method of three-dimensional (3-D) reconstruction of 2-D ultrasound images of the kidney is described. It is based on a coordinated spatial reconstruction of sequential cross-sectional images. The ultrasound head is moved longitudinally between two rails (parallel sections) and rotated. With a suitable computer program and contouring of each cross-section (so that the organ limits are defined for the computer) these cross-sectional pictures can be reconstructed into 3-D organ images. The kidney can then be presented spatially either as a binary picture or with closed surface. Ultrasound investigators are still unaccustomed to colour reproduction of 3-D reconstructed organs. It remains to be seen whether the method is valuable in routine clinical use. PMID- 3053087 TI - [Combined heart and kidney transplantation in terminal myocardial and renal insufficiency]. AB - In a 38-year-old man combined heart and kidney transplantation was performed successfully in one operation. Both organs have functioned well postoperatively: the patient was discharged from hospital on the 19th postoperative day and has remained in functional class I (New York Heart Association) eight months later. Only two episodes of cardiac rejection have been observed during this period, responding to treatment, and there was no evidence of rejection of the kidney. Such combined heart-kidney transplantation from one donor seems to be a promising form of treatment for patients in end-stage myocardial and renal failure. The frequency of cardiac rejections my be lower after combined heart-kidney transplantation than after cardiac transplantation alone. PMID- 3053088 TI - [Non-ulcer dyspepsia]. PMID- 3053089 TI - [Virus persistence--a key phenomenon of BVD virus infection]. PMID- 3053090 TI - [Metabolic overload tests in dairy cattle in evaluating performance ability]. PMID- 3053091 TI - [The effect of reciprocal translocations on the fertility of the mouse]. PMID- 3053092 TI - [Gene mapping in domestic animals--a mutual task of veterinary and animal breeding research]. PMID- 3053093 TI - [Sialic acid in boar sperm--the relation to qualitative and quantitative ejaculate parameters]. PMID- 3053094 TI - [The occurrence of toxin-forming Pasteurella multocida strains in the tonsil and nose of piglets from breeding stocks unsuspected of atrophic rhinitis]. PMID- 3053095 TI - [Rhinitis atrophicans (RA) of swine: a skin test for the detection of antitoxic Pasteurella multocida antibodies]. PMID- 3053096 TI - [The significance of domestic animals and pets as vectors of Campylobacter infections in humans]. PMID- 3053097 TI - [Calving with special attention to the natural process in regard to reproduction, milk production and animal rights]. PMID- 3053098 TI - [Fetlock joint flexion as the cause of a labor complication in a giraffe]. PMID- 3053099 TI - [Sudden "laying halt" as an adaptation reaction to heat stress]. PMID- 3053100 TI - [Indirect immunofluorescence--a rapid method for the detection of antibodies in viral diseases in the dog]. PMID- 3053101 TI - [Measuring the distribution of pressure on the bottom of cattle hooves- fundamental studies with a new type of measuring system]. PMID- 3053103 TI - [Different circadian influences in the administration of sulfur preparations on the metabolic liver functions]. PMID- 3053102 TI - [The honeybee (Apis mellifera) as an indicator of lead and cadmium levels in two locations]. PMID- 3053104 TI - [Improper feeding as stress]. PMID- 3053105 TI - ["The meat diet of humans" by Andreas Christian Gerlach (1875)]. PMID- 3053106 TI - [Mycobacteria in parrots and parakeets (Psittaciformes)]. PMID- 3053107 TI - [Single ovum triplets from an embryo transfer in cattle]. PMID- 3053108 TI - [The effect of treatment with depot-bromocriptine (CB 154-MK) on the hormone balance of the female dog]. PMID- 3053109 TI - [Growth hormone and insulin-like growth factor I (somatomedin C) blood levels in cattle from birth through sex maturation]. PMID- 3053110 TI - [Sonographic studies of the preovulatory follicle development in the mare]. PMID- 3053111 TI - [Natural mating in pastures of estrus-synchronized beef heifers with conspicuous fluctuation of the sex ratio of calves]. PMID- 3053112 TI - [The effect of diluents, dose size and freezing speed on the survival rate of deep frozen stallion sperm]. PMID- 3053113 TI - [The fluid balance in enteral salmonellosis of calves]. PMID- 3053114 TI - [The levels of blood cortisol in sheep during estrus and pregnancy]. PMID- 3053115 TI - [The insemination of horses in Neustadt-on-the-Aisch from 1980 to 1987]. PMID- 3053116 TI - [The relationship between different blood parameters as criteria for metabolic disorders and the milk cell count in dairy cows]. PMID- 3053117 TI - [The rumen pillars of domestic ruminants]. PMID- 3053118 TI - [The role of endocrinology in biotechnology]. PMID- 3053120 TI - [The effects of orally-administered therapeutic agents on the fermentation processes in the ruminal fluid of ruminating cattle. 1. Furazolidone]. PMID- 3053119 TI - [Intention tremor in a heifer with persistent BVD viremia]. PMID- 3053121 TI - [Comparative studies of the cryopreservation of mouse embryos with glycerin supplemented with sucrose or royal jelly]. PMID- 3053122 TI - [Successful embryo transfer in the horse]. PMID- 3053123 TI - Growth factor superfamilies and mammalian embryogenesis. AB - With the availability of amino acid and nucleotide sequence information has come the realization that growth factors can be clustered in to superfamilies. Several of these superfamilies contain molecules that were not initially identified because of growth-promoting activities; rather they were discovered through their ability to regulate other processes. Certain members of these superfamilies are present during early mammalian embryogenesis. However, until recently, it has been difficult to manipulate the developing mammalian embryo to observe directly the effects of inappropriate, excessive, or reduced expression of these molecules. Despite this limitation, at least some of these molecules have been implicated in the control of differentiation and morphogenesis, two actions unpredicted from the cell biology of most of the growth factors. Moreover, these actions are reflected in nonmammalian species where homologues of the mammalian growth factors control crucial steps in the choice of developmental fate. This review describes five growth factor superfamilies and the role these molecules may have in controlling proliferation, differentiation, and morphogenesis during mammalian development. PMID- 3053124 TI - Colloidal bismuth subcitrate. A review of its pharmacodynamic and pharmacokinetic properties, and its therapeutic use in peptic ulcer disease. AB - Colloidal bismuth subcitrate (CBS) possesses at least equal efficacy with histamine H2-receptor antagonist drugs in the treatment of peptic ulcer disease. However, CBS has the advantage of slower ulcer relapse rates than those seen after initial healing with the H2-antagonists. It has been postulated that this effect may be partly due to the antibacterial properties of CBS against Campylobacter pylori, a bacterium found in the gastric mucosa and gastric metaplasia within the duodenum of most patients with peptic ulcer and closely associated with gastritis. However, the role of C. pylori in the aetiology of peptic disease is far from clear. The mechanism by which CBS heals ulcers has not been fully elucidated, but several actions may be involved. CBS and mucus form a glycoprotein-bismuth complex in vitro. This provides a diffusion barrier to HCl and may, therefore, provide a protective coating in the ulcer crater which allows healing of the lesion to occur. Prostaglandin E2 production is also stimulated by CBS with subsequent secretion of alkali into the mucus layer. In addition, CBS has a direct inhibitory effect on C. pylori. Administration of CBS results in low levels of bismuth absorption. Most of the ingested bismuth is excreted as bismuth sulphide, causing blackening of the faeces, and the small amount absorbed is excreted in the urine. Bismuth intoxication (encephalopathy) has been reported with prolonged administration of bismuth salts, and there has been 1 report of similar intoxication in a patient receiving unusually high doses of CBS for a prolonged period. However, no such intoxication has been reported with CBS used at its recommended dosage in the acute treatment of peptic ulcer disease, and no other serious adverse effects have been associated with CBS. Tissue accumulation during prolonged therapy seems likely, and the safety of CBS during long term maintenance therapy has not been established. The lack of effect on gastric acid secretion is seen as an added advantage for CBS, since prolonged drug-induced hypochlorhydria has been postulated to have potentially detrimental effects. Thus, while the role of C. pylori in peptic ulceration requires further clarification, CBS would appear to have an important place in the treatment of peptic ulcer disease with the advantage of relatively slow relapse rates after initial healing and treatment discontinuation. PMID- 3053125 TI - Milrinone. A preliminary review of its pharmacological properties and therapeutic use. AB - Milrinone is a bipyridine derivative of amrinone, with approximately 10 to 75 times greater positive inotropic potency, and separate direct vasodilatory properties. As with amrinone, the relative importance of these properties to treatment of congestive heart failure still remain controversial. The mode of action of milrinone appears to be due in part to selective inhibition of a specific cardiac phosphodiesterase with a subsequent increase in intracellular cyclic adenosine monophosphate and alteration in intracellular and extracellular calcium transport. Clinical experience has involved both short and long term treatment of a limited number of patients with moderate to severe congestive heart failure refractory to conventional therapy. Milrinone has usually been administered as intravenous bolus doses (12.5 to 75 micrograms/kg) and/or continuous intravenous infusion (0.5 microgram/kg/min), or orally (30 to 40 mg/day in divided doses). Milrinone rapidly improves cardiac performance by enhancing myocardial contractility, and by decreasing systemic vascular resistance (afterload), left ventricular filling pressure (preload), and pulmonary arterial pressure. Exercise performance improvement occurs with enhancement of left ventricular performance but without a significant increase in myocardial oxygen consumption or significant decrease in mean arterial pressure. Milrinone has been compared with dobutamine, nitroprusside and captopril in preliminary short term studies in patients with severe congestive heart failure. Milrinone significantly increased stroke work index and decreased left ventricular filling pressure compared to nitroprusside. When compared with dobutamine, both drugs improved cardiac index (to a similar degree), but milrinone significantly reduced right atrial pressure, pulmonary capillary wedge pressure and left ventricular end-diastolic pressure. One small study suggests that short term effects of intravenous milrinone may be superior to those of oral captopril, and it appears that the addition of captopril to milrinone therapy may produce a synergistic haemodynamic effect. Preliminary long term studies suggest that tolerance to the haemodynamic effects of milrinone does not occur, and that the drug is well tolerated and without the thrombocytopenic effects, fever and gastrointestinal complications observed with amrinone. However, it has not been demonstrated that milrinone improves the prognosis of the disease or the overall mortality and its propensity to produce arrhythmias has not been fully agreed upon.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3053127 TI - Current management of malignant hypercalcaemia. AB - The pathogenic mechanisms causing malignant hypercalcaemia are primarily increased bone calcium mobilisation and renal calcium retention. In some reticuloendothelial malignancies, enhanced intestinal calcium absorption may also play a role. Malignant hypercalcaemia is a life-threatening condition, and there are many patients with malignancy in whom suppression of this complication is most desirable. In such cases, successful management of the hypercalcaemia will enable the overall treatment aims, such as tumour removal or ablation, to be achieved. Acute treatment involves the rapid lowering of serum calcium from potentially fatal concentrations, and comprises the use of intravenous rehydration, calcitonin and diphosphonates. In the longer term, other measures may be introduced to maintain and control the calcium concentration while specific antitumour therapy is instituted. PMID- 3053128 TI - Management of the pregnant diabetic patient. AB - The prognosis in diabetic pregnancy has greatly improved as a result of patient education and the availability of home blood glucose monitoring techniques enabling the implementation of good metabolic control pre-pregnancy, antenatal and intrapartum. These in turn have made possible the benefits to the offspring of vaginal delivery at term. Screening for gestational diabetes is important and the prognosis is also good where maternal normoglycaemia is achieved. All diabetic pregnancies should be cared for in specialist units under the supervision of an integrated team comprising an obstetrician, diabetologist and paediatrician, and for optimal results care should start prior to conception. PMID- 3053129 TI - [Lung compliance in chronic lung diseases]. PMID- 3053130 TI - [Who and what measures lung function in inflammation?]. PMID- 3053131 TI - [Continuation of tuberculosis research in the post-extinction era]. PMID- 3053132 TI - [Atypical mycobacteria causing generalized lung infections]. PMID- 3053133 TI - [The riddle of sarcoidosis]. PMID- 3053134 TI - [Organic dusts causing alveolar reactions]. PMID- 3053136 TI - [Diagnosis and treatment of sleep apnea]. PMID- 3053135 TI - [Modifications in the treatment of lung cancer]. PMID- 3053137 TI - [Is asthma also a disease of blood circulation in the respiratory tract?]. PMID- 3053126 TI - Fluoroquinolone antibiotics. Microbiology, pharmacokinetics and clinical use. AB - The newer fluoroquinolones are a major advance in antimicrobial chemotherapy. They inhibit the supercoiling activity of the DNA gyrase enzyme, thus exerting their antibacterial action on DNA and RNA synthesis, resulting in a biphasic response and killing of susceptible organisms. The newer fluoroquinolones have an extended antimicrobial spectrum compared to their older congeners, and are highly active against most Gram-negative pathogens including the Enterobacteriaceae and Pseudomonas aeruginosa. While Staphylococcus aureus and coagulase-negative staphylococci are usually susceptible to the fluoroquinolones, streptococci are generally more resistant and enterococci are resistant. All of the newer fluoroquinolones may be administered orally and most of them have been administered parenterally. They are widely distributed in the body, attaining therapeutic concentrations in most tissues. All of the fluoroquinolones have long half-lives and may be administered once or twice daily. The fluoroquinolones have proved effective in many infections, including uncomplicated or complicated urinary tract infections, respiratory tract infections, gonorrhoea, bacterial gastroenteritis, and soft tissue infections due to Gram-negative organisms. In general, success has been notable in the management of Gram-negative infections but less so with Gram-positive infections. Resistance has occurred and is proving a problem with P. aeruginosa in some cystic fibrosis patients, but as yet no plasmid-mediated resistance has developed. Cross-resistance occurs between the quinolones but only rarely with other classes of antibacterial drugs. The fluoroquinolones have an excellent safety record and their adverse effects, which include hypersensitivity reactions, dizziness, headache, gastrointestinal disturbance and minor haematological abnormalities are usually mild and transient. However, the fluoroquinolones have been found to damage juvenile weight-bearing joints in animals and are therefore only to be used with caution in children; transient arthralgia has been reported in a cystic fibrotic teenager on long term ciprofloxacin therapy. All of the fluoroquinolones except ofloxacin are associated with a variable increase in the serum concentration of theophylline, warfarin and caffeine. Thus, the fluoroquinolones are an attractive option in the management of many infections. Cost and possible resistance, however, should counsel caution in their use, and may limit them to situations where a cheaper antimicrobial of equivalent efficacy is not available. PMID- 3053138 TI - [New approaches in the pathogenesis of asthma affecting treatment patterns]. PMID- 3053139 TI - [Asthma induced by exercise and cold air]. PMID- 3053140 TI - [The mechanism of aerosol effects on the lung]. PMID- 3053142 TI - [The changing spectrum of occupational diseases of the lung]. PMID- 3053141 TI - [Chronic bronchial obstruction and the pathogenesis of emphysema]. PMID- 3053145 TI - End stage renal disease in tuberous sclerosis: a case report and review of the literature. PMID- 3053144 TI - [The respiratory tract and air pollution]. PMID- 3053143 TI - [Enclosure problems with the energy crisis]. PMID- 3053146 TI - Sclerotherapy in the treatment of bleeding oesophageal varices: preliminary report. PMID- 3053147 TI - Review of the environmental toxicity of quaternary ammonium halides. AB - This paper summarizes all available information on the environmental toxicity of quaternary ammonium compounds (QACs). Ten-minute contact kills of bacteria occur at 50-333 mg/liter. These chemicals are acutely toxic at approximately 1 mg/liter to invertebrates and fish and toxicity is occasionally as low as 0.1 mg/liter; no effect levels are generally 10 times lower than LC50 values. Toxicity to invertebrates and fish appears to be relatively independent of structure; the compounds studied, which have a large variety of structures, are all toxic at approximately the same order of magnitude. The QACs tested inhibited plant growth at 3-5 mg/liter. Predictions of toxicity to QACs are made based on knowledge of structure-activity relationships. PMID- 3053148 TI - [In vitro systems for the primary search for biologically active substances with carcinostatic action]. AB - Improvement and development of new methods for primary screening of antitumour drugs in vitro are based on the data on the points open to injury in the tumour cell metabolism and on the evidence of the known carcinostatic drug mechanism. Further development of the primary screening methods should proceed, probably, in two main directions. Further improvement of the methods for cloning tumour cells in the semifluid nutrient media is of great interest. On the whole the creation of new test systems is a significant trend of the scientific screening. PMID- 3053149 TI - [Gene rearrangement of immunoglobulins and of the T-cell receptor for antigens and its significance in the diagnosis of lymphoproliferative diseases]. AB - The blot-hybridization method for detecting gene rearrangement of Ig and T-cell receptor for antigen is briefly described. The results obtained by this method in various lymphoproliferative diseases offer new insights in the diagnosis of monoclonal lymphocyte expansion and in the determination of the lineage of proliferating cells. The ability of revealing oligoclonal lymphocyte proliferation and discriminating benign and malignant forms of angioimmunoblastic lymphadenopathy, T gamma-lymphocytosis, and some other disorders makes this method a useful tool for investigating diseases with lymphocyte proliferation. PMID- 3053150 TI - [Malignant lymphomas of various histological types in pike from the eastern portion of the Baltic Sea]. AB - Malignant lymphomas have been found in pikes (Esox lucius L.) caught in the eastern part of the Baltic Sea. The frequency of the disease has increased in recent years. Three different histological types of malignant lymphoma are described in 32 pikes: poorly differentiated diffuse lymphocytic lymphoma, mixed (histiocytoid-lymphocytic) diffuse lymphoma and undifferentiated (anisomorphic) diffuse lymphoma. PMID- 3053151 TI - Gastrointestinal angiodysplasia: clinical features. PMID- 3053152 TI - Angiodysplasia: morphologic diagnosis. PMID- 3053153 TI - Giant Brunneroma of the duodenum. AB - A case of giant hyperplasia of Brunner's glands in the duodenum is reported. Recurrent vomiting due to stenosis of the duodenum was the leading symptom. Endoscopy, ultrasound and histological examination were employed as complementary methods to establish the diagnosis. The 5 X 7 cm large tumor was surgically excised. Brunneromas are always benign and their incidence is about 0.08% in upper GI-tract endoscopy. PMID- 3053155 TI - Porous surfaced endosseous dental implants: fixation by bone ingrowth. PMID- 3053154 TI - Utilization, attitudes, and experiences of Vietnam Era veterans with Veterans Administration health facilities: the American Legion experience. AB - A random sample of American Legion members in six states who had served in the Armed Forces during the Vietnam Era was conducted through a mailed questionnaire, in order to determine patterns of usage of Veterans Administration health facilities, as well as attitudes toward the VA and experiences at these facilities. Of the 6810 male respondents, 42.0% had served in Southeast Asia. These subjects were categorized according to their level of combat in South Vietnam. Thirty-six percent of those who had served in Southeast Asia had used VA health facilities, compared to only 18% of men who served elsewhere. Among Southeast Asia veterans, combat level was an important predictor of extent of usage of VA facilities for problems of both physical and mental health. Combat level was also associated with lack of basic and major medical insurance. While men with lower incomes tended to make greater use of VA mental health facilities, nearly one-fourth of mental health users had family incomes above $30,000. Despite their greater usage of the VA, men with higher combat levels expressed lower feelings of security about this agency, and rated its staff less helpful and of lower quality than did men who experienced lower levels of combat. On the other hand, higher combat veterans thought themselves better informed about VA services. Men who had gone to the VA for mental health assistance reported a disturbingly low frequency of having been asked basic questions that relate to possible diagnosis of post-traumatic stress disorder (PTSD), questions related to combat, which may be one of its etiologic factors, or other questions relating to their military history. Because combat level in Vietnam veterans is a major determinant of both attitudes toward and utilization of VA health facilities, the VA as well as other health agencies which deal with Vietnam veterans should be especially sensitive to this factor, and should take it into consideration when evaluating veterans' physical and mental health. PMID- 3053156 TI - Dental implants: surgical considerations. PMID- 3053158 TI - Aorto-iliac thrombosis in two horses: clinical course of the disease and use of real-time ultrasonography to confirm diagnosis. PMID- 3053157 TI - Atypical myoglobinuria: an acute myopathy in grazing horses. AB - Four out of 12 horses grazing a field in Berkshire, England, suffered a prostrating illness and died within 12 to 72 h. Serum biochemical abnormalities, including markedly elevated muscle enzymes, were demonstrated and at post mortem widespread myodegeneration was found in both skeletal muscle and myocardium. Urine analysis revealed myoglobinuria, and renal changes were seen histologically. Although similar pathologically, the clinical syndrome and circumstances of the outbreak were not typical of equine exertional rhabdomyolysis (EER). The outbreak bore a striking resemblance to other reported sporadic outbreaks of an atypical myoglobinuria occurring in grazing horses. A number of potential aetiological and contributory factors (including herbicide toxicity) were considered, but the aetiology remains unresolved. PMID- 3053159 TI - Clonal sketches of the immune response. PMID- 3053161 TI - An initiation site of DNA replication with transcriptional enhancer activity present upstream of the c-myc gene. AB - We have previously reported that c-myc protein may promote cellular DNA replication by binding to initiation sites of replication. Here we report that a putative origin of human cellular DNA replication (ori) is present at approximately 2 kb upstream of the coding region of the c-myc gene itself. The c myc protein, or protein(s) complexed with c-myc protein, bind to the upstream region (approximately 200 bp in length) which has transcriptional enhancer activity as well as autonomously replicating activity in human cells, suggesting that the c-myc protein may be an enhancer binding protein as well as a DNA replication protein. Results with deletion mutants suggest that the sequence essential to the origin of DNA replication may be adjacent to, but cannot be clearly separated from, the sequence responsible for enhancer activity. Furthermore, when cloned DNA containing putative c-myc protein binding sequences was transfected as competitor into HL-60 cells, expression of c-myc was inhibited, suggesting that c-myc protein itself may be necessary for c-myc expression. PMID- 3053160 TI - Purification of a NF1-like DNA-binding protein from rat liver and cloning of the corresponding cDNA. AB - NF1-like proteins play a role in transcription of liver-specific genes. A DNA binding protein, recognizing half of the canonical NF1 binding site (TGGCA) present on the human albumin and retinol-binding protein genes, has been purified from rat liver. Several peptides deriving from a tryptic digest of the purified protein were sequenced and the sequence was used to synthesize specific oligonucleotides. Two overlapping cDNA clones were obtained from a rat-liver cDNA library; their sequence reveals an open reading frame coding for 505 amino acids, including all the peptides sequenced from the purified protein. The DNA-binding domain, most likely located within the first 250 amino acids, is highly homologous to the sequence of CTF/NF1 purified from HeLa cells. Northern analysis reveals several mRNA species present in different combinations in various rat tissues. PMID- 3053162 TI - A yeast DNA repair gene partially complements defective excision repair in mammalian cells. AB - The RAD10 gene of Saccharomyces cerevisiae is required for nucleotide excision repair of DNA. Expression of RAD10 mRNA and Rad10 protein was demonstrated in Chinese hamster ovary (CHO) cells containing amplified copies of the gene, and RAD10 mRNA was also detected in stable transfectants without gene amplification. Following transfection with the RAD10 gene, three independently isolated excision repair-defective CHO cell lines from the same genetic complementation group (complementation group 2) showed partial complementation of sensitivity to killing by UV radiation and to the DNA cross-linking agent mitomycin C. These results were not observed when RAD10 was introduced into excision repair defective CHO cell lines from other genetic complementation groups, nor when the yeast RAD3 gene was expressed in cells from genetic complementation group 2. Enhanced UV resistance in cells carrying the RAD10 gene was accompanied by partial reactivation of the plasmid-borne chloramphenicol acetyltransferase (cat) gene following its inactivation by UV radiation. The phenotype of CHO cells from genetic complementation group 2 is also specifically complemented by the human ERCC1 gene, and the ERCC1 and RAD10 genes have similar amino acid sequences. The present experiments therefore indicate that the structural homology between the yeast Rad10 and human Ercc1 polypeptides is reflected at a functional level, and suggest that nucleotide excision repair proteins are conserved in eukaryotes. PMID- 3053163 TI - Mitochondrial RNA polymerase of Saccharomyces cerevisiae: composition and mechanism of promoter recognition. AB - Mitochondrial RNA polymerase of Saccharomyces cerevisiae consists of two different proteins: a core RNA polymerase of 145 kd and a specificity factor of 43 kd, which contributes the capacity to recognize promoters of the various genes encoded in the mitochondrial genome. We purified both components by SDS-PAGE, followed by renaturation to the active state. The two components were used either singly or in combination to study their interactions with promoter-containing DNA fragments. The core component showed random and weak interaction with DNA, the specificity factor none at all, whereas both components together specifically bound to a promoter. In DNase I footprinting experiments, promoter-bound RNA polymerase protected a short region of DNA flanked by hypersensitivity sites and centred around the position at which RNA synthesis starts. The initial phase of transcription gave rise to specific changes in this footprint: the upstream border remained at the same position up to synthesis of a 4-nt RNA chain, whereas at the downstream border progressive disappearance of hypersensitivity sites took place. PMID- 3053164 TI - Expression of functional human EGF receptor on murine bone marrow cells. AB - The human epidermal growth factor-receptor (EGF-R) was introduced into primary mouse bone marrow cells (BMC), utilizing retrovirus mediated gene transfer. Cultivation of infected BMC in the presence of interleukin-3 (IL-3) led to the outgrowth of IL-3 dependent myeloid cells, which efficiently expressed functional EGF-R, exhibiting its two characteristic affinity states. EGF acts on these cells synergistically with IL-3 in stimulating DNA synthesis and cell proliferation even under IL-3 saturation conditions. However, EGF was not sufficient to replace the requirement for IL-3. In contrast, EGF was able to maintain proliferation of a factor-dependent hemopoietic cell line (FDC-P1) infected with the EGF-R retrovirus in the absence of IL-3, but these cells did not respond to EGF in the presence of IL-3. No influence of EGF on IL-3 induced mast cell differentiation of BMC expressing the EGF-R could be observed by histological criteria. These data show that the expression of EGF-R alone is not sufficient to induce or maintain cell proliferation in IL-3 dependent bone marrow derived cells, although it can do so in established hemopoietic cell lines. PMID- 3053165 TI - Multiple mechanisms regulate c-myc gene expression during normal T cell activation. AB - Quiescent normal human T cells express low levels of steady-state c-myc mRNA as a result of low constitutive promoter utilization, a block to transcriptional elongation within the gene, and rapid degradation of c-myc mRNA in the cytoplasm. Following the activation of the T cell receptor (TCR)/CD3 complex, quiescent T cells are induced to express c-myc mRNA. Two intracellular pathways, one involving protein kinase C activation and the other mediated by increased intracellular calcium concentration, are activated by TCR/CD3 receptor stimulation. These two pathways, which can be activated by phorbol myristate acetate (PMA) and ionomycin respectively, appear to play complementary roles in the transcriptional induction of c-myc gene expression by the antigen receptor complex. Ionomycin treatment of quiescent cells leads to enhanced c-myc expression primarily as a result of increased transcriptional initiation. In contrast, PMA contributes to c-myc expression, at least in part, by decreasing the block to transcriptional elongation present within the gene. Both the PMA- and ionomycin-mediated induction of c-myc expression can be independently enhanced by stabilization of c-myc mRNA in the cytoplasm. These observations demonstrate that multiple mechanisms co-operate to regulate c-myc gene expression during normal T cell activation. PMID- 3053166 TI - Sensitive and high resolution in situ hybridization to human chromosomes using biotin labelled probes: assignment of the human thymocyte CD1 antigen genes to chromosome 1. AB - A method for in situ hybridization originally developed for mapping genes in the nematode, Caenorhabditis elegans has been adapted for high resolution cytological mapping of genes in the human. The probe DNAs are labelled by incorporation of biotin dUTP and the site of hybridization detected by immunofluorescence. For the accurate assignment of the hybridization signal to chromosome bands, visualized by staining with Hoechst 33258, a heterologous ribosomal DNA probe is also included in the hybridization reaction. These rDNA signals are used as fiducial markers when aligning the two fluorescent images. We demonstrate the method by assignment of the human thymocyte CD1 antigen genes to human chromosome 1q22-23. PMID- 3053168 TI - Anaesthesia for combined pancreatic and renal transplantation and potassium homeostasis. AB - Diabetic patients who develop renal failure frequently have other serious manifestations of their disorder, including peripheral and central vascular disease, blindness, neuropathy and metabolic abnormalities which can include life threatening loss of those mechanisms that normally control potassium homeostasis. In this report, the anaesthetic and metabolic management of five diabetic patients with end-stage renal failure undergoing combined pancreatic and renal transplantation, is presented and discussed. PMID- 3053169 TI - Purification and characterization of the 90-kDa heat-shock protein from mammalian tissues. AB - The 90-kDa heat-shock protein (HSP90) has been purified from mammalian tissues, mouse liver and porcine brain, with a good yield by a new method involving hydrophobic chromatography. Mouse liver HSP90 and porcine brain HSP90 were compared with mouse lymphoma HSP90 which was purified from T lymphoma cell line, L5178Y, by a modification of the previously reported method. These three HSP90s were indistinguishable from one another in amino acid composition, one dimensional peptide mapping, elution pattern of proteolytic fragments (trypsin- or V8-protease-cleaved) in reverse-phase high-performance liquid chromatography, reactivity with the antibody against mouse T lymphoma HSP90 and the ability to bind to F-actin. The amino acid sequences of three portions (total 47 amino acid residues) of lymphoma HSP90 were determined and they were homologous to those of the corresponding portions of Drosophila HSP83A and yeast HSP90. These results suggest that HSP90 is a highly conserved protein during evolution. PMID- 3053167 TI - The promoter of Alzheimer's disease amyloid A4 precursor gene. AB - The promoter of the gene for the human precursor of Alzheimer's disease A4 amyloid protein (PAD gene) resembles promoters of housekeeping genes. It lacks a typical TATA box and shows a high GC content of 72% in a DNA region that confers promoter activity to a reporter gene in an in vivo assay. Transcription initiates at multiple sites. Sequences homologous to the consensus binding sites of transcription factor AP-1 and the heat shock control element binding protein were found upstream of the RNA start sites. Six copies of a 9-bp-long GC-rich element are located between positions -200 and -100. A protein--DNA interaction could be mapped to this element. The 3.8 kb of the 5' region of the PAD gene include two Alu-type repetitive sequences. These findings suggest that four mechanisms may participate in the regulation of the PAD gene and could be of relevance for the progression of amyloid deposition in Alzheimer's disease. PMID- 3053171 TI - Localization of the synthesis of very-long-chain fatty acid in mitochondria from Saccharomyces cerevisiae. AB - The localization of the mitochondrial elongation activities ('elongases') from Saccharomyces cerevisiae has been investigated. It was shown, using carboxyatractyloside in the incubation mixture, that synthesis of very-long-chain fatty acids probably occurred outside the matrix and, by fractionation experiments, that elongases are membrane-bound enzymes. The solubilization of the outer membrane by digitonin showed that three elongating activities are correlated with a marker of the outer membrane and not with an inner membrane marker. A further partial purification of the outer membrane showed that elongases are present in the outer membrane of mitochondria. PMID- 3053170 TI - Primary structures of Escherichia coli pyruvate formate-lyase and pyruvate formate-lyase-activating enzyme deduced from the DNA nucleotide sequences. AB - The structural gene of pyruvate formate-lyase (pfl) and that of pyruvate-formate lyase-activating enzyme were shown to be adjacent on the chromosomal map of Escherichia coli. DNA sequencing was performed along a stretch of 3592 nucleotides to obtain the amino acid sequences of both proteins. The derived primary structures (759 and 245 residues) were confirmed by partial structure analyses on the purified proteins. The open reading frames are separated by a 194 nucleotide stretch, and their flanking regions include signal elements that are compatible with separate control of protein synthesis from the two genes. PMID- 3053172 TI - Isolation and sequence analysis of the fatty acid synthetase FAS2 gene from Penicillium patulum. AB - The fatty acid synthetase complex of Penicillium patulum was isolated and shown to be structurally similar to other known fungal fatty acid synthetases. It is composed of two subunit, alpha and beta, each with a molecular mass of about 200 kDa. P. patulum genomic and cDNA libraries were constructed in lambda gt11 and EMBL3 vectors. From these libraries, the P. patulum FAS2 gene together with its flanking DNA was isolated. The cloned genomic DNA was sequenced over a length of 6357 base pairs. The coding sequence of fatty acid synthetase subunit alpha, being 5571 nucleotides long, was identified within this DNA segment. The FAS2 gene is a mosaic of three exons (514, 4949 and 108 base pairs) and two introns, each of 54 base pairs in length. Both introns were absent in the corresponding cDNA sequences. Like other fungal introns both contain an internal CTAAC sequence, located 10 base pairs upstream of their 3'-exon/intron boundaries. In addition, they have, at their ends, the GTCAAGT and TAG consensus sequences characteristic of all eucaryotic introns. Furthermore, two pairs of direct repeats, of as yet unknown significance, were found in the two P. patulum introns. The P. patulum FAS2 gene encodes a protein of 1857 amino acids and 204.5 kDa molecular mass. It is 90 nucleotides shorter than the corresponding S. cerevisiae gene. In both organisms, the FAS2 genes and their products exhibit a high degree of overall sequence similarity at both the DNA (63%) and protein (68%) levels. Therefore, the fatty acid synthetase alpha subunits of P. patulum and S. cerevisiae obviously contain the same catalytic domains in an identical sequential order. PMID- 3053173 TI - Mutants with base changes at the 3'-end of the 16S RNA from Escherichia coli. Construction, expression and functional analysis. AB - The functionally important 3' domain of the ribosomal 16S RNA was altered by in vitro DNA manipulations of a plasmid-encoded 16S RNA gene. By in vitro DNA manipulations two double mutants were constructed in which C1399 was converted to A and G1401 was changed to either U or C and a single point mutant was made wherein G1416 was changed to U. Only one of the mutated rRNA genes could be cloned in a plasmid under the control of the natural rrnB promoters (U1416) whereas all three mutations were cloned in a plasmid under the control of the lambda PL promoter. In a strain coding for the temperature-sensitive lambda repressor cI857 the mutant RNAs could be expressed conditionally. We could show that all three mutant rRNAs were efficiently incorporated into 30S ribosomes. However, all three mutants inhibited the formation of stable 70S particles to various degrees. The amounts of mutated rRNAs were quantified by primer extension analysis which enabled us to assess the proportion of the mutated ribosomes which are actively engaged in in vivo protein biosynthesis. While ribosomes carrying the U1416 mutation in the 16S RNA were active in vivo a strong selection against ribosomes with the A1399/U1401 mutation in the 16S RNA from the polysome fraction is apparent. Ribosomes with 16S RNA bearing the A1399/C1401 mutation did not show a measurable protein biosynthesis activity in vivo. The growth rate of cells harbouring the different mutations reflected the in vivo translation capacities of the mutant ribosomes. The results underline the importance of the highly conserved nucleotides in the 3' domain of the 16S RNA for ribosomal function. PMID- 3053174 TI - Ultrastructural localization of anionic sites on the surface of yeast, hyphal and germ-tube forming cells of Candida albicans. AB - The cell wall of Candida albicans contains chitin, beta-glucans and phosphorylated mannoproteins, and possesses a fuzzy coat which is thought to play a role in pathogenicity, phagocytosis, and adherence of this dimorphic yeast. Using scanning electron microscopy and the gold method, mannoproteins were detected on the whole surface of blastoconidia including the bud scars, but chitin was absent even after alpha-mannosidase treatment of the cells. The presence of surface beta-(1----6)glucan (but not beta(1----3)glucan) was observed only after extensive alpha-mannosidase and alkaline phosphatase treatments of blastoconidia. Using transmission and scanning electron microscopy, the locations of anionic sites were revealed by polycationic colloidal gold-chitosan complexes on the surface of blastoconidia, germ tubes and hyphae. Anionic sites were dispersed evenly over the surface of blastoconidia bearing bud scars. Depending upon the growth conditions, anionic sites could be detected on emerging buds and young cells. However, bud scars were always free of marking. When germ-tube formation was induced, anionic sites were present at different densities on all cell surfaces, the highest density being observed on cells with bud scars. Anionic sites were detected at a remarkably high density on all hyphal surfaces. An apical concentration of anionic sites was observed on germ tubes and hyphae. The distribution of anionic sites was not modified by endoglucosaminidase treatment of blastoconidia, germ tubes and hyphae. The anionic sites were associated with the fuzzy coat. As the hyphal form is regarded as possessing the greatest invasiveness, it is suggested that anionic sites play an important role in establishing tissue colonization by this human pathogen. PMID- 3053175 TI - Nucleolar and nuclear envelope proteins of the yeast Saccharomyces cerevisiae. AB - We have developed a fast and reliable purification protocol to obtain yeast nuclei in intact and pure form and in a reasonable yield. The purified nuclei appear homogeneous at the light and electron microscopic level, are highly enriched in the nuclear marker histone H2B and devoid of mitochondrial, vacuolar and cytosolic marker proteins. On sodium dodecyl sulfate (SDS)-polyacrylamide gels, the nuclear fraction contains unique proteins which distinguishes them from the major yeast subcellular fractions. Yeast nuclei were separated by detergent/salt extraction into soluble, insoluble and membrane fractions. Antibodies raised against subnuclear fractions lead to the identification of an integral nuclear membrane protein and a high-abundance 38-kDa protein which is located in the yeast nucleolus. PMID- 3053176 TI - Randomized controlled trial of low dose warfarin in the primary prevention of ischaemic heart disease in men at high risk: design and pilot study. AB - We report the pilot stage of a double-blind randomized controlled trial of low dose warfarin in the primary prevention of ischaemic heart disease (IHD) in men at high risk. The first objective was to see if levels of factor VII coagulant activity, VIIc, could be reduced without undue difficulty from a mean level of about 115% to about 70% (the level in patients on conventional warfarin doses being about 30%). This was accomplished with a mean daily dose of 4.6 mg warfarin. The international normalized ratio (INR) corresponding to a VIIc value of 70% was about 1.5. The second objective was to assess the risk of bleeding associated with the intended level of anticoagulation. There was no significant excess in the number of actively treated men ever reporting nose bleeds, possible haematuria, rectal bleeding or bruising, although there may have been an increase in the frequency of rectal bleeding in men who did report this symptom. The third objective was to establish the willingness of patients to take part in a trial of this kind. Of those invited to the initial screening examination, 72% attended. Of those invited to enter the treatment phase of the trial, 71% did so. Compliance with trial treatment was at a very high level. The rate of withdrawal from randomized treatment was within acceptable limits, at about 15% over a three or four-year period. The scientific case for a full trial is strong and the pilot trial shows that it could be accomplished. PMID- 3053178 TI - The treatment of chronic pulmonary hypertension by vasodilators: hope and hesitation. AB - Contemporary therapy of most advanced cases of chronic lung disease is not yet satisfactory; even long-term oxygen treatment has shortcomings and limitations. Many authors have tried to find another therapeutic approach for the correction of either pulmonary hypertension or oxygen delivery, both factors being related to long-term survival. Vasodilators seem to be the subject of further intensive research rather than an established therapy. The results of acute experiments with different drugs are promising but studies of long-term treatment are incomplete. Some of the most pertinent questions for further research are: the classification of the reversibility of pulmonary hypertension, reliable evaluation of the results of therapy, the application of animal experiments to human pathology, haemodynamic conception of an ideal vasodilator, and the attenuation of the effect of some vasodilators during long-term treatment. PMID- 3053177 TI - Acute neurogenic pulmonary oedema following generalized tonic clonic seizure. A case report and a review of the literature. AB - A case of acute neurogenic pulmonary oedema following a generalized tonic clonic seizure is presented. The condition is rare and the exact pathophysiology is unknown. Pulmonary oedema is a serious complication of epileptic seizures, however, and the importance of awareness of this condition and its treatment is emphasized by a high mortality rate in older epileptics and by the demonstration of pulmonary oedema in recent series of young epileptics who died suddenly. PMID- 3053179 TI - Experimental prevention of hypoxic pulmonary hypertension in animals by drugs. AB - The rat model of chronic hypoxic pulmonary hypertension has been extensively studied and shows many of the features seen in man with chronic pulmonary hypertension. The development and reversibility of these changes by various treatments and by drugs is discussed. The experimental model may provide valuable clues as to the mechanisms involved in the aetiology of pulmonary hypertension in man. PMID- 3053180 TI - Natural variability of pulmonary haemodynamics. AB - In the interpretation of the effect of a drug or intervention in a single patient, it is desirable to know the spontaneous fluctuations of the variable under study. A few studies have determined pulmonary haemodynamic variability in acute and short-term intervals. It seems reasonable to adopt a confidence limit of 6 mmHg or 22% for PPA changes to be significant. The long-term evolution is difficult to determine because of the unavoidable patient selection for repeat examinations several years apart, and because of the necessary treatment. In slight or moderate pulmonary hypertension the changes in PPA with time seem to be of small magnitude: less than 1 mmHg per year. PMID- 3053181 TI - Haemodynamic requirements for an ideal pulmonary vasodilator. AB - An ideal vasodilator should be selective for the pulmonary vascular bed, thus minimizing side-effects from reduced systemic resistance. It must achieve not only a drop in pulmonary vascular resistance but also a marked decrease in pulmonary arterial pressure. The ideal drug should increase cardiac output and pulmonary venous oxygen saturation. An increase in oxygen delivery to the peripheral tissues should be achieved at a lower right ventricular afterload both at rest and during exercise. Right ventricular function should be improved. The effects of vasodilator therapy should be so marked that it should be possible to follow them non-invasively by radionuclide methods and exercise tolerance tests. The aim of vasodilator therapy is a regression of pulmonary hypertension and of right ventricular hypertrophy, an improved quality of life, and above all a longer survival. PMID- 3053182 TI - Clinical trials with long-term treatment of pulmonary hypertension due to lung disease. AB - Many drugs have been tested for the treatment of pulmonary hypertension of different origins, but even the acute effects are controversial and side-effects are frequently important. In a few studies, vasodilators have been used in the long-term treatment of chronic pulmonary hypertension related to chronic lung disease (calcium channel blockers, hydralazine, nitrates); here, also, the results are inconstant and the side-effects are sometimes major. No vasodilator specific for the pulmonary circulation is presently available. PMID- 3053183 TI - The United States experience with the acute and chronic treatment of primary pulmonary hypertension. AB - Patients who clinically have primary or 'unexplained' pulmonary hypertension are found at autopsy or lung biopsy to have a variety of pulmonary vascular changes, including medial hypertrophy, thrombosis, intimal fibrosis and plexiform lesions. It is not surprising that the haemodynamic response to vasodilators varies widely. In general, the non-specific vasodilators used to treat pulmonary hypertension cause an acute fall in systemic arterial pressure, with an increase in cardiac output and a reduction in pulmonary vascular resistance. Pulmonary arterial pressure usually does not change much but occasionally drops dramatically. The risk of death in an acute trial of a vasodilator is less than 0.5% in experienced hands. The use of a short-acting vasodilator (e.g., prostacyclin) may indicate the presence or absence of vasoconstriction, the likelihood of fixed structural obstruction to flow and the risk of administering longer-acting vasodilators, and it may give a clue to prognosis. The risk-benefit ratio in the use of vasodilators in the long-term treatment of primary pulmonary hypertension needs to be evaluated by a controlled trial, conducted in those who respond acutely. The role of high-dose calcium channel blocker treatment and multiple drug therapy will also require further study. PMID- 3053184 TI - Current trends in research and clinical trials of drugs for chronic pulmonary hypertension. PMID- 3053185 TI - Morphological substrate for the reversibility and irreversibility of pulmonary hypertension. AB - The reversibility of pulmonary hypertension in hypertensive pulmonary vascular disease depends, in the first place, on the feasibility of eliminating the cause of the elevation of pressure. Equally important in this respect are the type, the severity, and the extent of the pulmonary vascular lesions. This implies that various forms of pulmonary hypertension have completely different tendencies for regression. In thrombotic arteriopathy, whether caused by primary thrombosis or by embolism, the chance of regression of pulmonary hypertension, and of the vascular lesions, is limited. On the other hand, in many patients pulmonary venous hypertension and the associated vasculopathy, even when severe, appear potentially reversible. Experimental evidence suggests that the same is true in cases of hypoxic pulmonary hypertension as long as prominent complicating vascular alterations, as often observed in chronic obstructive lung disease, are absent. In plexogenic arteriopathy regression of pulmonary hypertension, following elimination of its cause, is observed whenever the vascular lesions have not progressed beyond a certain stage that can be considered a point of no return. Thereafter, there is not only no regression but a distinct tendency to progression of pulmonary vascular disease. PMID- 3053186 TI - The stability of 99mTc-DTPA and 99mTc-HIDA following ultrasonic nebulisation. AB - The stability of 99mTc-DTPA (diethylenetriamine-pentaacetic acid) and 99mTc-HIDA (2,6-diethylacetanilido-iminodiacetic acid) were evaluated following ultrasonic nebulisation. The results confirm that either of these radiopharmaceuticals can be used in our nebuliser without significant radiochemical breakdown. PMID- 3053187 TI - Hyperthermia in cancer therapy. AB - Tumor hyperthermia is a rediscovered technique of oncotherapy which has confirmed value in many studies on cell cultures, rodent and mammalian tumors as well as first investigations on patients with tumors. The biological basis for using heat in the treatment of cancer is well established. Various direct and indirect mechanisms are significant for the effect of hyperthermia on tumor tissue. Whereas there are already extensive studies on the direct effects of hyperthermia on DNA, RNA, and protein synthesis, energy metabolism, and the membrane properties of tumor cells, the indirect effects have only been investigated more closely in recent years. These are likewise important for the damage to the tumor tissue and are mediated above all via alterations in the microcirculation and the environment. The recently gained increasing significance of this new technique in combination with other treatment modalities is well documented. Technical problems of heat application must be overcome, especially in deeper tumors and problems of thermometry must be solved in order to be able to apply tumor hyperthermia not only to selected advanced or recurrent tumors, but in order to use it as the fourth pillar of tumor therapy besides surgery, radiotherapy and chemotherapy. This article considers the biological basis and important aspects of hyperthermia therapy in combination with radiotherapy and chemotherapy. PMID- 3053188 TI - Rubella vaccination. AB - Rubella vaccination programmes aim to prevent congenital rubella infections. Previously differing programmes have now converged according to the following principle: First vaccination should be given at the age of 15 months (together with measles and mumps vaccine) to both boys and girls, in order to diminish the circulation of the wild virus. Teenage girls require (re-)vaccination to ensure their immunity. Also non-immune women have to be identified and vaccinated before they become pregnant. A low acceptance rate increases the risk of infection of pregnant women, independent of the vaccination omitted. As a rule natural and vaccine-induced immunity prevents congenital rubella infections. These infections are exceedingly rare in children born to immune mothers, and are always caused by the wild virus. This minimal risk will disappear only with the eradication of rubella virus, still a distant goal in countries offering vaccination only on a voluntary basis. PMID- 3053189 TI - The effect of human growth hormone therapy on skinfold thickness in growth hormone-deficient children. AB - Skinfold thickness (ST) was measured in 43 children with various forms of growth hormone (GH) deficiency during the first year of GH therapy. The average (and SEM) initial ST, expressed as standard deviation score (SDS) was 1.17 (0.25) for subscapular, 0.63 (0.18) for triceps, and 0.40 (0.21) for biceps ST. During therapy the average decrease is 1 SD. Children in the pubertal age group and those with partial GH deficiency showed smaller decreases. A larger decrease of triceps ST was associated with lower GH and insulin peaks, and lower age, bone age and initial weight-for-height. Some correlations between ST decrease and growth response in the first year were significant, but still too low to allow of reliable predictions. The same was true for other clinical parameters. These data indicate that a chronic lack of GH leads to unequal fat distribution, possibly due to different sensitivities to GH in the trunk and extremities. The variability of ST responses to GH therapy limits clinical applications. PMID- 3053190 TI - Immunocytochemical detection of neuroblastoma cells infiltrating clinical bone marrow samples. AB - To evaluate the feasibility and clinical usefulness of immunocytochemical detection of bone marrow metastases in neuroblastoma, we studied bone marrow samples from patients undergoing intensive therapy, followed in the majority of cases by autologous bone marrow rescue. Two monoclonal antibodies were used in an indirect immunoenzymatic assay to test 384 samples collected from multiple bone marrow sites during 79 staging procedures in 48 patients. Of 578 immunocytochemical tests, 59 (10%) yielded non-evaluable results. Analysis by individual bone marrow sites showed an agreement between cytological and immunocytochemical examinations in 276 of 309 (89%) evaluable tests with 5 A7 and in 179 of 210 (85%) with UJ 13 A. Infiltration by neuroblastoma cells was reported in 9% of samples by cytology, in 6% by immunochemistry with 5 A7 and in 16% with 13 A. Analysis of results by staging demonstrated agreement between cytological examination and immunocytochemical detection with both monoclonal antibodies in 60 of 75 (80%) evaluable stagings. Bone marrow metastasis was detected by cytology in 22% of stagings, by immunochemistry with 5 A7 in 23%, with UJ 13 A in 25%. Detailed analysis of discordant results revealed that they were related partly to bone marrow sampling variability associated with focal and minimal metastasis of neuroblastoma cells. These data suggest the clinical usefulness of immunocytochemical detection as a complementary test to cytological examination for accurate evaluation of bone marrow infiltration in patients with disseminated neuroblastoma. PMID- 3053191 TI - A modified nitrite test as a screening test for significant bacteriuria. AB - The presence of nitrite was initially assessed in 306 specimens of urine. If the initial result was negative a sample of urine was incubated at 37 degrees C. The other portion was also incubated but with the addition of one drop of 1% NaNO3, and the nitrite test was then repeated. The results were correlated with the results of quantitative routine urine culture. The modified test significantly (P less than 0.01) improved the sensitivity (93% v 21%), and the predictive negative value (97% v 80%). PMID- 3053192 TI - Survival of a premature neonate with obstructive anuria due to Candida: the role of early sonographic diagnosis and antimycotic treatment. AB - A low birth weight premature neonate with systemic candidiasis developed complete renal obstruction by fungus balls, diagnosed by ultrasonography. The neonate was treated with temporary urinary diversion, amphotericin B, 5-fluorocytosine and survived. This case emphasizes the need for a high index of suspicion of renal obstruction by fungus balls in neonates with systemic candidiasis when renal function deteriorates. In such cases early urinary diversion can be life-saving. PMID- 3053194 TI - Tumor-specific antigen for human renal cell carcinoma. Ultrastructural localization of the antigen by immunoelectron microscopy. AB - Hybridoma technology enabled the production of tumor-specific monoclonal antibodies reactive exclusively with human renal cell carcinoma. Intracellular localization of the antigen was undertaken in order to gain understanding of its possible physiological role in cellular metabolism and to investigate its future clinical applicability for immunoscintigraphy in tumor localization. For immunoelectron microscopy a special paraformaldehyde-periodate fixation process (PLP fixation) had to be employed, in order to preserve the cell's ultrastructure without destruction of the antibody-binding epitope. The antigen was found to be strictly intracytoplasmic in close correlation to the glycogen particles characteristic for human renal cell carcinoma. These findings suggest that this antigen may be involved in the pathological glycogen synthesis, explaining the specific staining pattern of these monoclonal antibodies. PMID- 3053193 TI - Gender differences in age at onset of schizophrenia. An overview. AB - We present the results of a review of the literature concerning gender differences in age at the onset of schizophrenia. In view of the very consistent finding that the first admission to hospital for schizophrenia occurs on average earlier in men than in women we examined the question whether this is due to the fact that the psychosis manifests itself earlier in men or that the period between first manifestation and admission to hospital is shorter than in women. By means of a metaanalytic approach we then looked for evidence for the existence of local or temporal variations in the degree of gender difference. Lastly, we dealt with the question whether gender differences in age of onset can be observed in other functional psychoses. PMID- 3053195 TI - Disorders of voiding and bladder function presented in textbooks published during the nineteenth century. PMID- 3053196 TI - Tuberculosis in cadaveric renal allograft recipients. Report of 4 cases and review of the literature. AB - Tuberculosis is an uncommon infectious complication after kidney transplantation, but such an infection carries a significant mortality and morbidity and may jeopardize the function of the renal allograft. Herein we report our experience in the treatment of mycobacterial infections in 4 renal transplant recipients and review the literature. PMID- 3053197 TI - Leiomyosarcoma of the ureter. AB - Primary ureteral tumors occur relatively infrequently compared to tumors of the kidney and bladder. Smooth muscle tumors of the ureter are very rare and only 9 cases of leiomyosarcoma have been reported in the literature. The treatment of choice is total nephroureterectomy with en bloc excision of the tumor. Prognosis is unfavorable as indicated by the short survival of the reported cases. PMID- 3053198 TI - Experimental studies on the heterogeneity of bladder carcinoma. PMID- 3053199 TI - Chemotherapeutic management of invasive bladder carcinoma. PMID- 3053200 TI - Chemotherapy in the management of invasive and metastatic bladder cancer. PMID- 3053201 TI - Precursor lesions of invasive bladder cancer. PMID- 3053203 TI - Thymus and immunity--I. Early thymus research. AB - This paper is a review of early results in thymus research that found their full significance after several decades, when the strategic role of the organ in immunology was recognized. PMID- 3053202 TI - The control of hormone-dependent breast cancer growth--are we talking about estrogen alone? PMID- 3053204 TI - Thymus and immunity--II. The last three decades. PMID- 3053205 TI - Use of selected combinations of monoclonal antibodies to tumor associated antigens in the diagnosis of neoplastic effusions of unknown origin. AB - While conventional cytodiagnosis can, in most instances, recognize cancer cells in metastatic effusions from solid tumors, the cellular type or the organ of origin of the primary neoplasia can rarely be determined only on the basis of their morphology. In the present study we have evaluated whether immunocytochemical techniques can be used to overcome this limitation by employing a panel of monoclonal antibodies (MoAbs) to tumor associated antigens (TAA) which lack detectable reactivity with mesothelial cells. To this end we have analyzed, by indirect immunofluorescence, cytospins of 60 malignant effusions of unknown origin. The results of this study have shown that the definition of the origin of the primary tumor, which was subsequently confirmed histologically and/or clinically, could be reached in 87% of the cases. These findings demonstrate that selected combinations of MoAbs, when used in immunocytochemical tests, can provide a powerful diagnostic tool in defining the site of cryptic primary neoplasias causing metastatic effusions. PMID- 3053206 TI - Comparative effect of cisplatin, spiroplatin, carboplatin, iproplatin and JM40 in a human myeloid clonogenic assay. AB - The relative toxicities of cisplatin and its analogs, spiroplatin, carboplatin, iproplatin and JM40, were tested against normal human progenitor myeloid cells (CFU-GM) in a clonogenic assay. Cells obtained from five bone marrows were incubated for 60 min with various drug concentrations and plated. The mean inhibitory concentrations for 50% of the bone marrow colonies (IC50) were 15.6 micrograms/ml for cisplatin, 0.4 micrograms/ml for spiroplatin, 56.3 micrograms/ml for carboplatin, 36.3 micrograms/ml for iproplatin and 179.5 micrograms/ml for JM40. Ratios of the IC50s of the analogs with cisplatin as reference drug closely followed the corresponding ratios of the clinical maximum tolerated doses. This correlation between the CFU-GM assay results and the clinical myelotoxicity suggests that the assay is adequate for predicting myelotoxicity in vitro and selecting in vitro drug concentrations for the human tumor clonogenic assay. PMID- 3053208 TI - Post-operative intrapleural BCG in lung cancer. PMID- 3053207 TI - Comparative toxicity of cisplatin, carboplatin (CBDCA) and iproplatin (CHIP) in combination with cyclophosphamide in patients with advanced epithelial ovarian cancer. AB - Sixty patients with FIGO stage IIb, IIc, III and IV ovarian cancer were entered into a randomized Phase III study of cyclophosphamide 600 mg/m2 with cisplatin 100 mg/m2, iproplatin 240 mg/m2 or carboplatin 300 mg/m2. Dose modifications were made according to renal function and myelotoxicity. The arms containing carboplatin (CBDCA) and iproplatin (CHIP) were not shown to be significantly different from the cisplatin containing arm with regard to response rate, duration of response and survival. Subjective toxicity showed that cisplatin and cyclophosphamide therapy was associated with more nausea and vomiting (P = 0.0005). The duration of vomiting showed a significant increase with successive courses of chemotherapy for the cisplatin containing arm only (P less than 0.003). The cyclophosphamide/CHIP combination caused significantly more diarrhoea (P less than 0.0006). Alopecia was more severe (P less than 0.02), and neurotoxicity was more common, in patients who received cyclophosphamide and cisplatin (paraesthesiae P = 0.0007, tinnitus P less than 0.00005, deafness P = 0.0018). All three combinations caused cumulative toxicity on haemoglobin (Hb) (P less than 0.001 for each treatment), leukocyte count (WCC) (P less than 0.0005 for each treatment), and platelet count (P less than 0.0005 for each treatment). The degree of fall in Hb for each course of therapy was greater in the cisplatin containing arm compared with the CHIP and CBDCA arms which were not significantly different from each other (P = 0.0005). For WCC the cisplatin/cyclophosphamide regimen was significantly less toxic than CHIP/cyclophosphamide, with CBDCA/cyclophosphamide falling between the two and not being significantly different from either (P = 0.0005). The CHIP containing arm caused more thrombocytopenia than the other arms which were of equal toxicity (P less than 0.0005). Serum creatinine showed a gradual significant overall rise with each course of cisplatin/cyclophosphamide therapy (P less than 0.0005), whereas the CBDCA arm showed no change and the CHIP arm showed a small fall in serum creatinine after most courses of therapy. This study showed that CHIP or CBDCA in combination with cyclophosphamide was less toxic than cisplatin/cyclophosphamide therapy with regard to alopecia, degree and duration of nausea and vomiting, renal toxicity, neurotoxicity and anaemia. The CHIP/cyclophosphamide regimen caused more thrombocytopenia and diarrhoea. The CHIP and CBDCA containing arms caused more leukopenia than the cisplatin containing regimen. Either iproplatin or carboplatin would be an acceptable alternative to cisplatin in chemotherapy regimens, and would result in reduced toxicity. PMID- 3053209 TI - The pharmacokinetics of ceftazidime in patients with impaired renal function and concurrent frusemide therapy. AB - We have studied the pharmacokinetics of ceftazidime in 37 patients suffering from serious bacterial infections. All the patients had impairment of renal function and received moderate to high doses of frusemide concurrently. The doses of ceftazidime were given according to renal function as recommended by the manufacturer. Serum and urine samples were frequently collected, and drug concentrations measured by high performance liquid chromatography. The patients were grouped and evaluated according to renal function, mean (SD) creatinine clearances ranging from 70.1 (12.4) to 11.0 (3.2) ml.min-1. The pharmacokinetics of ceftazidime depended on renal function. A statistically significant increase in ceftazidime elimination half-life and decreases in urinary recovery, total body clearance, and renal clearance in proportion to the decrease in renal function were observed (p less than 0.05). The apparent volume of distribution also increased, but not significantly (p greater than 0.05). A linear correlation was found between the total body and renal clearances of ceftazidime and creatinine clearance. The extrarenal clearance increased from 3.9 to 14.0 ml.min 1 with decreasing renal function. Concurrent treatment with ceftazidime and moderate to high doses of frusemide did not impair renal function and no evidence of nephrotoxicity was found. PMID- 3053210 TI - Spiramycin does not increase plasma cyclosporin concentrations in renal transplant patients. PMID- 3053211 TI - Effect of fibrinopeptides A and B on human and rat platelet aggregation in vitro. AB - The effect of fibrinopeptides on platelet aggregation is reported. Fibrinopeptide A (minimal effective concentration, 0.65 microM) aggregated human (but not rat) platelets suspended in plasma and at lower concentrations (0.01-0.1 microM) potentiated platelet aggregation due to ADP and collagen in both species. Fibrinogen mimicked these effects of fibrinopeptide A. P-bromophenacyl bromide (100 microM), mepacrine (10 microM), indomethacin (10 microM) and dazoxiben (10 microM) inhibited human platelet aggregation induced by fibrinopeptide A and fibrinogen. In both species, fibrinopeptide B (0.65-6.5 microM) antagonised the platelet inhibitory effect of PGI2 and PGD2 but not adenosine. Antagonism was non competitive in nature. The concentration of fibrinopeptide A required to potentiate platelet aggregation occurs naturally in the plasma of patients with thrombotic disease suggesting this effect may be of physiological significance during the formation of a thrombus. The novel action of fibrinopeptide B to reduce the platelet inhibitory effect of PGI2 and PGD2 may also contribute to the control of thrombus formation. PMID- 3053212 TI - EEDQ, a tool for ex vivo measurement of occupancy of D-1 and D-2 dopamine receptors. AB - EEDQ (N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline) inactivates dopamine (DA) D 1 and D-2 receptors, measured by ex vivo [3H]SCH 23390 and [3H]spiperone binding in striatal homogenates. SCH 23390 (D-1 antagonist) and SK&F 38393 (D-1 agonist) protect against inactivation of D-1 receptors (ED50 values 0.075 and 53 mumol/kg, respectively). Raclopride (D-2 antagonist) and quinpirole (D-2 agonist) protect against inactivation of D-2 receptors (ED50 values 0.48 and 7.1 mumol/kg, respectively). The potencies correspond closely to those obtained by in vivo binding experiments using radioligands. PMID- 3053213 TI - Mandibular rotations: concepts and terminology. PMID- 3053214 TI - Cephalometric standards for the southern Chinese. PMID- 3053215 TI - A comparison of DNA and immunoblot fingerprinting of the SII biotype of coagulase negative staphylococci. AB - Thirty-eight isolates of the SII biotype of coagulase negative staphylococci were examined by phase typing, antibiograms, DNA and immunoblot fingerprinting. Multiple isolates were available from 5 patients with clinically proven infections and from 12 further patients who were epidemiologically distinct. Only 3 of the isolates were phage typable and antibiograms produced only 9 types. All isolates were typable by both DNA and immunoblot fingerprinting and reproducibility was excellent provided the conditions were standardized. Discrimination was better with immunoblot fingerprinting (17 types) than DNA fingerprinting (11 types). Both techniques were successful in assessing the significance of multiple isolates from a single patient. PMID- 3053216 TI - Incidence and nature of human tuberculosis due to bovine tubercle bacilli in South-East England: 1977-1987. AB - A total of 201 new cases of tuberculosis due to bovine tubercle bacilli was confirmed in South-East England between 1977 and 1987 inclusive. This represents about 1% of all cases of tuberculosis in this region. Most cases occurred amongst older individuals of indigenous white British origin, although some younger patients of Southern European and Indian subcontinent ethnic origin were also diagnosed. The lung was the most frequent site of disease, followed by the genito urinary tract. In view of the known risk of transmission of disease from man to cattle via the respiratory and urinary tracts, continued surveillance of this relatively uncommon form of tuberculosis is still indicated. PMID- 3053217 TI - A two-year survey of the incidence of heat-labile enterotoxin-producing Escherichia coli and other enteric pathogens in travellers returning to the Sheffield area. AB - A case-controlled study of the incidence of heat-labile enterotoxin-producing Escherichia coli (LT+ETEC) and other enteric pathogens in travellers returning to the Sheffield area was conducted from May 1984 to April 1986. LT+ETEC were found in 35 (5.8%) of 600 travellers to developed countries (mainly popular Mediterranean holiday resorts), 36 (11.3%) of 320 travellers to less-developed countries, and 11 (0.9%) of 1282 control patients whose illness was not associated with recent travel abroad. A seasonal peak of LT+ETEC infection was observed only in travellers to developed countries, with infections being significantly commoner in August to October. There was no significant deviation from expected age/sex distribution of LT+ETEC infection. Strains of LT+ETEC from travellers produced more toxin than strains from control patients, strains from travellers to less-developed countries producing most of all. PMID- 3053218 TI - Occurrence of 'thermophilic' campylobacters in sewage and their removal by treatment processes. AB - Removal of thermophilic campylobacters from sewage at three different stages of treatment at a trickling filter sewage works has been assessed. Samples of incoming sewage, primary sedimentation effluent and final effluent were taken daily from 06.00 h to 20.00 h for 5 consecutive days and the numbers of campylobacters determined by using a most probable number method. Each sample was cultured using 2 h pre-enrichment followed by enrichment in Preston broth for 48 h and detection by plating. Over 78% of the incoming campylobacters were removed after primary sedimentation and less than 0.1% remained in the final effluent. Campylobacter jejuni biotype I and biotype II constituted 81.5% and 15.9% respectively of the 232 isolates tested. Serotypes common in sewage were common in human faeces. It appears that the trickling filter sewage works removes most of the campylobacters entering the sewage works, but large numbers, estimated to be approximately 10(10), are released into the environment daily from a local sewage works. PMID- 3053219 TI - Use of colistin and sorbitol for better isolation of Serratia marcescens in clinical samples. AB - A comparison was made of different culture media and procedures for detection of Serratia marcescens from faecal, pharyngeal and ocular swabs collected from 213 neonates. MacConkey agar and MacConkey agar with sorbitol (1%) and/or colistin (200 i.u./ml) were used both for primary isolation and after enrichment using Mossel Enterobacteriaceae broth with colistin (200 i.u./ml). The use of MacConkey agar supplemented with colistin for primary isolation improved considerably the isolation rate of S. marcescens from faecal swabs but not from pharyngeal swabs; the number of ocular isolations were insufficient to demonstrate differences between procedures. Moreover the enrichment procedures consistently increased the number of S. marcescens isolates especially from pharyngeal and ocular swabs. Use of sorbitol made detection of S. marcescens from clinical specimens easier and time- and cost-efficient. PMID- 3053220 TI - The use of sorbitol-MacConkey agar in conjunction with a specific antiserum for the detection of Vero cytotoxin-producing strains of Escherichia coli O 157. AB - Using DNA probes specific for the genes encoding Vero cytotoxins 1 and 2 in hybridization experiments on faecal samples, Vero cytotoxin-producing Escherichia coli (VTEC) of serogroup O 157 were detected in 21 of 63 cases of haemorrhagic colitis, 9 of 31 cases of non-bloody diarrhoea and 14 of 68 cases of haemolytic uraemic syndrome. Compared with these results sorbitol-MacConkey agar in conjunction with a specific O 157 antiserum gave a sensitivity of 62% in haemorrhagic colitis, 56% in non-bloody diarrhoea and 57% in haemolytic uraemic syndrome. The specificity of this method was 100% in all three groups. This demonstrates that sorbitol-MacConkey agar is a useful screening method for the detection of VTEC of serogroup O 157 when used in conjunction with a specific homologous antiserum. However, this method does not detect VTEC belonging to other serogroups and such strains were found, particularly in cases of haemolytic uraemic syndrome. PMID- 3053221 TI - Seroepidemiology of group B Streptococcus type II antibody specificity. AB - The specificity of human antibodies for the two major sidechain determinants of the type II group B streptococcal (GBS) polysaccharide was examined in 90 pairs of maternal and cord sera. Using an ELISA system, total antibody was measured against the complete (sialylated) type II antigen and the proportion of antibody against the galactose determinant was estimated by inhibition with free beta methylgalactopyranoside. Mothers colonized by type II or by other GBS types had higher levels of total specific antibody (means, 3.3 and 4.7 micrograms/ml, respectively) than those not colonized (mean, 2.2 micrograms/ml). Cord sera averaged 1-2 micrograms/ml lower than maternal sera. Colonization with GBS was also associated with higher levels against the galactose determinant (mean, 1.5 micrograms/ml, compared to 0.7 micrograms/ml for those not colonized). The distribution of specificities favoured antibodies against the sialic acid determinant in maternal but not cord sera. Specificity as well as antibody level may play a role in the epidemiology of GBS type II. PMID- 3053222 TI - Chlamydial antibodies in farmers in north-west England. AB - Because of recent reports of abortion in farmers' wives following infection with ovine strains of Chlamydia psittaci during pregnancy, the distribution of chlamydial antibodies was studied in rural populations in north-west England, where endemic chlamydial infection with abortion is common in sheep. Immunoperoxidase assays with C. trachomatis and ovine C. psittaci showed no significant differences in either the frequency or titres of antibodies between sheep farmers and other types of farmer or non-farming adults living in the same areas. The frequency and titres of antibodies in farmers' wives were no greater than in farmers, and were unrelated to their previous obstetric history or type of farming. Overall, 62/255 (24%) of this rural population had antibody detected by C. trachomatis antigen and only 30/255 (13%) detected by C. psittaci antigen. The possible significance of these findings is discussed. This survey does not suggest that the risk of infection with C. psittaci is especially high in people working with sheep, but the complications following infection during pregnancy deserve the specific instructions that have been given to pregnant women to avoid exposure, especially during lambing, in farming and veterinary work. PMID- 3053223 TI - Maternally derived measles immunity in children of naturally infected and vaccinated mothers. PMID- 3053224 TI - Human corneal endothelial cells: isolation, characterization and long-term cultivation. AB - Long-term cultures of human corneal endothelial cells have been established. In culture, these cells form a dense monolayer (about 500,000 cells cm-2), similar to that found in vivo, and synthesize an extracellular matrix containing laminin, entactin, and fibronectin. Factor VIII and angiotensin-converting enzyme were not found in either the cultured or native corneal endothelium. Cells were obtained by scraping corneal buttons that had been preincubated in the culture medium supplemented with endothelial cell mitogen. The human corneal endothelium was grown under conditions virtually the same as those used for cultivation of human vascular endothelial cells, namely, on fibronectin- or gelatin-coated tissue culture plastic in Medium 199 supplemented with 20% human serum and 400 micrograms ml-1 endothelial cell growth supplement. Human corneal endothelial cells from the culture obtained can be used for transplantation onto human corneas, for studying repair of damaged corneal endothelium in situ, as well as for in vitro studies of cell growth regulation. PMID- 3053225 TI - Crystallins and their synthesis in human lens epithelial cells in tissue culture. AB - Explants of epithelial cells from young human lenses of 5-12 months of age, obtained from patients who underwent surgery for retinopathy of prematurity, were cultured in Dulbecco's modified Eagle's medium supplemented with 20% fetal calf serum. Without exception, every piece of the anterior capsule explant showed cell outgrowth within 48-72 h and resulted in confluent monolayer culture within 2 weeks. From these monolayer cultures, two to three passages of subcultures were obtained by routinely seeding cells in a ratio of 1:4. The doubling times for these human lens epithelium (HLE) cultures during the first 4 weeks of two passages were found to be 24-36 h. In a majority of cultures through the first three passages, more than 12 population doublings were attained. However, no lentoid bodies were formed during this period. These cells were studied for the presence of crystallins and their synthesis. Using SDS-polyacrylamide gel electrophoresis, the presence of alpha- and beta-crystallins was demonstrated in HLE cells through three passages. The amount of alpha-crystallin in the first two passages amounted to nearly 13% of the total protein, but decreased significantly in the third passage. The presence of crystallins was corroborated by antibody reaction to the specific crystallins. Indirect immunofluorescence revealed the presence of actin and vimentin in these cell cultures. The synthesis of crystallins in HLE cultures was shown by the incorporation of [35S]methionine which was time dependent. The crystallin synthesis was found to decrease in third passage when the cell growth slowed down without consistent formation of confluent monolayer. These studies have demonstrated that primary cultures of HLE cells can be successfully grown from young lenses through several passages which continue to express the characteristic crystallins of the epithelial cells. PMID- 3053226 TI - Excitatory amino acids and rod photoreceptor disc shedding: analysis using specific agonists. AB - L-Glutamate and L-aspartate stimulate photoreceptor disc shedding. In order to evaluate the possible involvement of a receptor, we examined the effects of specific excitatory amino acid agonists on rod photoreceptor disc shedding and neural retinal toxicity. Using eyecups from both Xenopus laevis and Rana pipiens, we found that kainate, quisqualate, and N-methyl-D-aspartate (NMDA) were all neurotoxic, but that kainate caused a more extensive inner retinal lesion. Kainate also caused disc shedding at concentrations as low as 10 microM; dihydrokainate, a structural analogue, was at least 100-fold less potent. In contrast, quisqualate induced disc shedding only at concentrations above 5.0 mM, and NMDA had no effect on disc shedding at any concentration examined. Our results suggest that excitatory amino acids act via a receptor of the kainate type to effect disc shedding. The mechanism in the retina or photoreceptor pigment epithelial complex by which an excitatory amino acid receptor system influences disc shedding remains to be identified. PMID- 3053227 TI - Laminin distribution during corticospinal tract development and after spinal cord injury. AB - The glycoprotein laminin is a prominent constituent of basal laminae and has been suggested to play an important role in axonal growth. We have tested this hypothesis, by examining the temporal and spatial distribution of laminin in the rat spinal cord, relative to elongating corticospinal tract (CST) axons, during normal development and after newborn and adult spinal lesions. The distribution of laminin was demonstrated in spinal cord sections from animals ranging in age from 14 days embryonic to adult using immunocytochemistry. Anti-laminin immunolabeling was seen around blood vessels and meninges in all the animals examined. However, within the grey and white matter its distribution was age dependent. In the normal cord, immunostaining appeared in small amounts in early embryos, but was absent from all postnatal animals even at ages when the CST was growing down the cord. Following injury, intense immunostaining was associated with lesions in both newborn and adult operates at all postoperative periods examined. Within the matrix of the lesion laminin immunostaining was especially prominent. In the intact cord it was prominent only around blood vessels near the lesion site. Our results indicate that the distribution of laminin does not closely correlate with axonal growth of the CST either during normal development or after spinal injury. PMID- 3053228 TI - Establishment of Schwann cell cultures from adult rat peripheral nerves. AB - Schwann cell cultures are difficult to obtain from adult rat because of the abundant amount of connective tissue and myelin. We have developed a method for isolation and culture of these cells by mechanical and chemical dissociation which represents a modification of a previously described procedure for human neurofibromas. Schwann cells were identified in indirect immunofluorescence by the capacity to bind antibodies to S-100, galactocerebroside, and laminin. The baseline cell proliferation index was assessed by immunofluorescence of bromodeoxyuridine. This method provides Schwann cells from adult rat nerves for at least 7 days and in sufficient numbers for (a) morphological and immunological characterization and (b) analysis of the effects of mitogenic factors. PMID- 3053229 TI - The use of primary cultures of adult rat hepatocytes to study induction of enzymes and DNA synthesis: effect of nafenopin and electroporation. AB - Primary cultures of adult rat hepatocytes maintained in a well-differentiated state, in a chemically defined medium containing 2% DMSO, have been utilized to study the effect of non-mutagenic hepatocarcinogens such as the peroxisome proliferator nafenopin. The parameters chosen in this in vitro system were those that paralleled the major in vivo effects of nafenopin on the liver, mainly: the proliferation of the endoplasmic reticulum and induction of cytochrome P-452, the proliferation of the peroxisome compartment and the induction of cyanide insensitive beta-oxidation of fatty acids and the stimulation of liver growth as measured by the DNA synthetic activity of the hepatocytes. In this review, we also describe the morphology of hepatocyte cultures prepared from previously electroporated hepatocytes and the potential for the use of electroporation to introduce growth related genes into hepatocyte cells to study the mechanisms of hepatocyte growth at the molecular level. In addition we describe the formation of endoplasmic reticulum whorls in these cultures as a consequence of nafenopin treatment. 'Whorl formation' by hepatotrophic chemicals has been previously shown to occur in vivo; in this report, it is described for the first time in vitro. PMID- 3053230 TI - Cellular models for the detection and evaluation of drugs that modulate human phagocyte activity. AB - Simple testing models have been developed for the evaluation of chemical or biological compounds that modulate the activity of human phagocytes. Human neutrophils from buffy coats of donor blood are used. They are stimulated with receptor agonists, and the effects of test compounds on exocytosis of different enzymes, the generation of superoxide (respiratory burst), and cytotoxicity are quantified. All assays are performed in microtiter plates and the responses are evaluated by multi-well photometry or fluorimetry. The models are apt to detect compounds acting on phagocytes as agonists or antagonists, signal transduction activators or inhibitors and primers of agonist responses, and to assess cytotoxic effects. PMID- 3053232 TI - Cryopreservation of parasites. AB - In this review, advances in cryopreservation of helminth parasites are reported. Our own studies demonstrate that metacestodes of Echinococcus multilocularis can be maintained in a viable state for at least 1-2 years by appropriate deep freezing and storage in liquid nitrogen. Infective larvae of the nematode Toxocara canis cryopreserved for 1 week in liquid nitrogen were maintained after thawing in vitro in a chemically defined medium for 35 weeks. Although motility of previously deep-frozen larvae was reduced they produced secretory/excretory antigens of similar immunodiagnostic quality as those from unfrozen larvae. Whereas infective larvae of several species of trichostrongy-lids can be easily cryopreserved, the infective larvae of the cattle lungworm, Dictyocaulus viviparus, and muscle larvae of Trichinella spiralis are more sensitive to damage by subzero temperatures. Therefore, survival rates after cryopreservation are low, but improvement of the cooling schedules appears to be feasible. It is concluded that cryopreservation of certain stages of helminth and protozoan parasites is a useful technique for long-term storage of defined isolates, which can contribute considerably to reducing the number of experimental animals usually required for serial passages. PMID- 3053231 TI - Development of DNA probes for cytotoxin and enterotoxin genes in enteric bacteria. AB - DNA probes to identify the genes encoding toxins in enteric bacteria have been developed. Use of these probes reduces the number of animals required for toxicity testing, as suspect bacteria can be directly tested for the presence of toxin. We have augmented the gene probes available by developing probes against the Escherichia coli enterotoxin LTII and shiga toxin from Shigella dysenteriae 1. The LTII gene from E. coli 357900 was identified and characterised and a suitable internal probe was obtained. The LTII gene was found not to be common among enterobacteriae from various geographical locations. Isolates predominately of animal origin from Nigeria and Thailand hybridized with the probe. The shiga toxin gene was isolated from S. dysenteriae 1 by a combination of in vivo and in vitro methods. An internal probe was identified and used against different serogroups of Shigella and E. coli isolates. The probe was found to hybridize with S. dysenteriae 1 isolates and also some S. flexneri and S. sonnei strains. Representatives were tested for toxin production and found to produce toxin at low levels. PMID- 3053233 TI - The use of microtiter plates for the simple and sensitive determination of insulin by an ELISA method. AB - A method of insulin determination using a commercially available ELISA kit was modified for use in microtiter plates. The adapted assay, based on the binding of porcine anti-guinea pig insulin antibodies to microtiter plates and insulin peroxidase conjugate as displacer, is sensitive between 0.5 and 30 ng/ml. Since it uses only 10-40 microliter of sample material it enables the determination of 5-100 pg of insulin. The rapid (5-6 h), automatable, reproducible and reliable assay makes it possible to determine many samples in a short time. PMID- 3053234 TI - Interaction of benznidazole reactive metabolites with nuclear and kinetoplastic DNA, proteins and lipids from Trypanosoma cruzi. AB - Epimastigotes of Trypanosoma cruzi (Tulahuen strain Tul 0 stock) biotransform benznidazole (N-benzyl-2-nitro-1-imidazole acetamide) to reactive metabolites that bind covalently to DNA, proteins and lipids of the parasite. These effects might be related to the trypanocidal action of benznidazole, a chemotherapeutic agent against Chagas' disease. PMID- 3053235 TI - Electrophoretic protein typing of Campylobacter jejuni subspecies "doylei" (nitrate-negative campylobacter-like organisms) from human faeces and gastric mucosa. AB - Twenty-seven strains comprising 23 clinical isolates of nitrate negative campylobacters (NNC) from Australia, South Africa, the United Kingdom and the Federal Republic of Germany, a representative of the CNW (catalase negative/weak) group and reference strains of three other Campylobacter species, were characterized by one-dimensional SDS-polyacrylamide gel electrophoresis of cellular proteins. The protein patterns were highly reproducible, and were used as the basis for a numerical analysis which showed that the reference strain (NCTC 11951) of Campylobacter jejuni subspecies "doylei", and 20 NNC isolates formed a distinct group at the 74% similarity level. The protein patterns showed unexpectedly low similarity between subspecies "doylei" and the type strain of Campylobacter jejuni and revealed that some NNC strains were quite distinct from subspecies "doylei". Four electrophoretic (EP) types (I-IV) were identified from phenons formed at the 81% similarity level. Three of these (I, III, IV) corresponded to geographical location of strain isolation but the type II strains were from diverse locations. The correlation observed between EP-type, catalase production and sensitivity to 2, 3, 5, triphenyltetrazolium chloride indicated these latter two tests might be useful for biotyping within the subspecies. PMID- 3053237 TI - Antimicrobial resistance patterns and plasmids of enteropathogenic Escherichia coli isolated in Nigeria. AB - In an epidemiological study of enteropathogenic Escherichia coli, 102 strains were isolated from patients seen at the University Teaching Hospital in Lagos. The most common serotype encountered was 055 followed by 026. Antimicrobial susceptibility testing and plasmid profiling of the strains were done. All the strains were sensitive to colistin, nalidixic acid, nitrofurantoin, cefotaxime, amikacin, and augmentin. Of the 102 strains, 47 (46%) were resistant to one or more of the following antimicrobial agents: Co-trimoxazole, tetracycline, ampicillin, streptomycin, sulphonamide and a combination of ampicillin with sulbactam. All the strains that were resistant to any antimicrobial agents were also resistant to tetracycline. Seventy-two strains (70.6%) harbored plasmid whose molecular weights ranged from 0.8 to 120 x 10(6) daltons. The majority of the plasmid were smaller than 6 x 10(6); 90% of strains carrying plasmid ranging in size from 2 to 6 x 10(6) daltons and 50 to 70 x 10(6) daltons were resistant to one or more antimicrobial agents. Transformation and conjugation experiment showed that about 57% of the resistant strains carried R plasmid. Plasmid determined resistance to tetracycline, ampicillin, streptomycin and sulphonamide was found. PMID- 3053239 TI - Current treatment of hairy cell leukaemia. PMID- 3053238 TI - R-plasmids in Salmonella isolates from sporadic cases of gastroenteritis. AB - Five-hundred and twenty seven strains of Salmonella isolated from different patients admitted to hospitals in Rome from 1982 to 1985 were screened for their resistance to antimicrobial drugs. Sixty-one strains (11.6%) were found to be resistant to two or more antibiotics; the most frequent resistances were to sulfathiazole, streptomycin, tetracycline, chloramphenicol and ampicillin. Of the thirty-eight strains showing resistance to three or more antibiotics, 17 were able to transfer their resistance to E. coli K 12. The isolates were heterogeneous in plasmid population: only few strains harbored a sole plasmid, most harbored many plasmids ranging between 20 and 120 megadaltons in weight. Most strains were found to carry a conjugative plasmid of incompatibility group Inc H of 100-120 megadaltons and Inc I alpha of 60-70 megadaltons. PMID- 3053236 TI - Molecular aspects of the phagocytosis resistance of group A streptococci. PMID- 3053240 TI - Alkaline phosphatase-positive B cell lymphomas. AB - Alkaline phosphatase (ALP) activity of 70 cases of non-Hodgkin's lymphomas of the B-cell type was studied. ALP activity was found in malignant lymphoma (ML), follicular, small cleaved cell (1/5 cases); ML, diffuse, small cleaved cell (3/13 cases); and mantle zone lymphoma (intermediate lymphocytic lymphoma) (2/2 cases). The ALP-positive neoplastic cells simultaneously displayed the characteristic immunophenotype of mantle zone (MZ) B lymphocytes of secondary follicle (SIg D+, BA-1+, IL-2R+ and Leu-1+). All other B-cell lymphomas, including ML, follicular, mixed small cleaved and large cell (9 cases); ML, follicular, large cell (4 cases); ML, mixed small and large cell (7 cases); ML, diffuse, large cell (27 cases); and ML, small noncleaved cell (3 cases), were consistently negative for ALP. The present study indicates that ALP-positive lymphomas including follicular lymphomas and diffuse lymphomas may be neoplastic counterparts of ALP-positive MZ B lymphocytes. PMID- 3053241 TI - T-cell depletion versus methotrexate as GvHD-prophylaxis in allogeneic bone marrow transplantation for leukaemia. AB - Graft-versus-host disease (GvHD) prophylaxis using methotrexate (23 patients) and T-cell depletion of the graft (40 patients) was compared in 63 allogeneic bone marrow transplantations (BMT) for leukaemia. T-cell depletion significantly reduced (p = 0.001) the incidence of GvHD from 68% to 11% and the GvHD-associated mortality from 79% to 5%. Actuarial disease-free survival for low-risk patients (57% with T-cell depletion and 47% with MTX) was not significantly improved, due to graft failure and possibly due to a higher leukaemic relapse rate after T-cell depletion. Prevention of graft failure after T cell-depleted BMT is essential and could also reduce the risk of leukaemic relapse by improved engraftment. PMID- 3053242 TI - A case of chronic neutrophilic leukemia with original chromosomal abnormalities. AB - We report a new case of the unusual myeloproliferative syndrome chronic neutrophilic leukemia (CNL) that met all the criteria generally required for the diagnosis of this entity. The patient presented abnormalities in platelet function not previously reported that may explain the bleeding tendency observed in these patients. The study of neutrophil function suggested also defective mobility and intracellular bactericidal activity. The chromosomal study revealed original abnormalities consisting of multiple chromosomal ruptures and figures. The disease was controlled with busulfan. After 20 months, the patient died of sepsis. An autopsy was performed confirming the diagnosis and ruling out the existence of a cause of a leukemoid reaction, such as cancer or granulomatous disease. PMID- 3053243 TI - Thermodynamic parameters of the binding of the tight-binding I12X86 lac repressor to operator and non-operator DNA. AB - The thermodynamic parameters delta H and delta S corresponding to the binding of the tight-binding double mutant lac repressor I12X86 with operator and non operator DNA fragments were determined using the nitrocellulose filter binding assay. In both cases the binding processes are entropically driven and accompanied by an unfavorable enthalpy variation. The differences between these parameters and those previously reported for the wild type lac repressor show that the strategy adopted by the mutant to interact with DNA is highly different from that of the wild type repressor and suggest more hydrophobic contacts between the mutant and DNA. PMID- 3053245 TI - Cloning a synthetic gene for human stefin B and its expression in E. coli. AB - A gene coding for human stefin B was synthesized by the solid-phase phosphite method and cloned in the pUC8 cloning vector. The insert with the verified DNA sequence was subcloned into two expression vectors and expressed in E. coli as a fusion protein with beta-galactosidase and as a native protein. The CNBr cleaved fusion protein and the native recombinant stefin B were inhibitory to papain and reacted with antibodies against human stefin B. PMID- 3053244 TI - Proteinase yscE of yeast shows homology with the 20 S cylinder particles of Xenopus laevis. AB - Proteinase yscE of the yeast Saccharomyces cerevisiae has been compared with the 20 S cylinder particles of Xenopus laevis. Both proteins are characterized by a similar group of 10-12 polypeptides with molecular masses ranging between 21 and 38 kDa. Antibodies generated against the 20 S Xenopus cylinder particles show cross-reactivity with yeast proteinase yscE subunits. The Xenopus particles and yeast proteinase yscE exhibit an identical image in electron microscopy. Both proteins appear as hollow cylinders mostly composed of four stacked annuli. The Xenopus 20 S particles exhibit proteolytic activity against the three peptide derivatives known to be substrates of proteinase yscE. The pH optimum for activity and the inhibition spectrum of the proteolytic activities of Xenopus 20 S particles and of yeast proteinase yscE are identical. The RNA content of the cylinder particles and of proteinase yscE is below 0.1 RNA chain per molecule. Our data suggest that proteinase yscE from yeast and the 20 S cylinder particles of X. laevis are homologous, highly conserved proteins carrying the catalytic character of a peptidase. PMID- 3053246 TI - Nuclear Overhauser effects in aqueous solution as dynamic probes in short linear peptides. AB - The possibility of obtaining interresidue NOEs from short linear peptides in aqueous solution has been investigated from an experimental point of view using peptides of various lengths (namely GGRA, LHRH and RNase S-peptide). It is shown that, provided that long (approximately 800 ms) NOESY mixing times are used, complete sets of sequential alpha N NOEs are obtainable. From the intensities and signs of the observed NOEs, the relative mobilities of different parts of the polypeptide chain can be determined. PMID- 3053247 TI - Photo-CIDNP study of the interaction between lac repressor headpiece and lac operator DNA. AB - Lac repressor headpiece (HP) and intact lac repressor have been studied using the photo-CIDNP method. At neutral pH histidine 29, tyrosines 7, 12 and 17 and methionine 1 are polarised. His-29 polarizations are weaker and broader in HP59 than in HP51 indicating that the C-terminal octapeptide in HP59 adopts a conformation that allows an interaction with His-29. The photo-CIDNP spectra of intact lac repressor and HP51 are very similar, showing that the same residues are accessible to the photo-excited flavin. An equimolar mixture of HP51 and a 14 base pair lac operator fragment strongly suppresses the photo-CIDNP effect of tyrosines 7 and 17 and abolishes the His-29 polarizations. The results are compared with earlier photo-CIDNP measurements on a complex of headpiece with poly[d(AT)] and with a model derived from a 2D NMR study on a lac headpiece operator complex. PMID- 3053248 TI - UV-induced cross-linking of proteins to plasmid pBR322 containing 8-azidoadenine 2'-deoxyribonucleotides. AB - An efficient method of cross-linking DNA to protein is described. The method is based on the incorporation of photoactive 8-azidoadenine 2'-deoxyribonucleotides into DNA. We have found that 8-N3dATP is a substrate for E. coli DNA polymerase I and that 8-N3dATP can be incorporated into plasmid pBR322 by nick-translation. Subsequently we were able to cross-link a set of different proteins to 8-azido-2' deoxyadenosine-containing pBR322 by UV irradiation (366nm). No DNA-protein photocross-linking was observed under the same conditions when the non photoactive plasmid pBR322 was used. PMID- 3053249 TI - Expression of human cathepsin B protein in Escherichia coli. AB - A cDNA fragment containing the coding sequence for the mature enzyme of human lysosomal proteinase cathepsin B was inserted in the pET plasmid expression vectors, so that it was placed under the control of transcription and translation signals from bacteriophage T7. Upon induction, cathepsin B antigen was detected by in situ immunoscreening of lysed E. coli and by Western blot analysis of bacterial lysates. To our knowledge this is the first report of abundant synthesis of cloned cathepsin B in any expression system. Subfragments of cathepsin B can also be generated by this technique and will be used to study cathepsin B structure and function. PMID- 3053251 TI - Suture or graft? Changing trends in melanoma wound closure. AB - The method of wound repair following excision of primary cutaneous malignant melanoma has been assessed in a consecutive series of 256 melanomas of the trunk and limbs between 1972 and 1986. Excision margins of 1, 2 and 3-5 cm were used according to clinical assessment of tumour thickness. Primary closure was the preferred method of wound repair and split skin grafting when it could not be achieved. 30% of wounds were closed primarily between 1972-81, rising to 54% between 1982-86. This change has been partly due to an increase in the number of thin lesions, but also due to improved surgical technique. Simple interrupted suture was used between 1972-81. In 1982 a multilayer subcutaneous and subcuticular prolene suture was introduced as an improved method of direct closure, and led to a reversal in the ratio of grafting to primary closure. This technique is particularly beneficial for wounds with excision margins of 2 and 3 cm. Complication rates have been lower for primary suture (4%) than split skin graft (12%). PMID- 3053250 TI - Cytosolic ADP enhances the sensitivity to tolbutamide of ATP-dependent K+ channels from pancreatic B-cells. AB - The effects of intracellular purine nucleotides on tolbutamide-induced block of ATP-dependent K+ channels from mouse pancreatic B-cells were studied using the patch-clamp technique. When applied to the inside of excised patches, tolbutamide alone blocked channel activity half-maximally at 55 microM and the concentration response curve for the inhibition of K+ channels by tolbutamide was flat. ADP (1 mM), but not other nucleotides (AMP, GTP or GDP) increased the steepness of the concentration-response curve and decreased the half-maximally effective tolbutamide concentration to 4.2 microM. It is suggested that the ATP-dependent K+ channel or a closely related structure contains a receptor which is accessible for cytosolic ADP and controls the sensitivity to tolbutamide. PMID- 3053252 TI - [Pseudotuberculosis]. PMID- 3053253 TI - [65th anniversary of the Union of Red Cross Societies and of the Red Crescent of the USSR]. PMID- 3053254 TI - Therapeutic hysteroscopic procedures. AB - Hysteroscopy has evolved from a diagnostic procedure into a therapeutic method for a variety of conditions. Instruments specifically designed for hysteroscopic operative procedures have improved. The indications for therapeutic hysteroscopy are increasing and its proper applications can improve patient's gynecologic care. These facts should stimulate the gynecologist to become proficient with hysteroscopy for diagnosis and the treatment of many intrauterine abnormalities. In selected patients there are major advantages including the avoidance of a laparotomy with the potential sequelae that can follow operations requiring entrance into the peritoneal cavity. The operative techniques have been refined and, with experience, good postoperative results and low morbidity have become evident. The septate uterus can be treated by hysteroscopic metroplasty with improved reproductive outcome. Symptomatic submucous myomas in selected patients can be removed hysteroscopically. Lysis of intrauterine adhesions has become the standard method of therapy. Foreign bodies lost in the uterine cavity or embedded IUDs can be removed atraumatically under direct vision. As the approach to intrauterine problems is refined, other applications are being investigated such as tubal cannulation, endoscopic chorionic villus sampling, and application of hysteroscopy to new reproductive technologies such as the placement of gametes in the fallopian tubes. Because of the simplicity of approaching the uterotubal ostia transcervically, hysteroscopy remains in the front line of investigation as a possible approach for inducing tubal occlusion as a permanent or temporary method of contraception. Finally, laser energy is being used in patients with intractable uterine bleeding to photocoagulate the endometrium and to create amenorrhea by means of inducing severe intrauterine adhesions. PMID- 3053255 TI - Endocrine profile of follicles containing oocytes with subsequent polyploid fertilization. AB - In an attempt to identify oocytes at risk for polypronuclear fertilization, follicular fluids were obtained retrospectively that contained oocytes that fertilized normally and abnormally. Whenever possible, each patient served as her own control during the same stimulation cycle. Twenty-six of 169 patients had oocytes that became polypronuclear, and of those 26, 21 had oocytes that fertilized and cleaved normally. Follicular fluids were analyzed for estradiol, progesterone, androstenedione, transferrin, and insulin. Insulin levels were noted to be significantly elevated (P less than 0.05) in the polypronuclear group when compropose that insulin, a known growth factor for granulosa cells cultured in vitro, when present in excessive concentrations may predispose to polypronuclear fertilization. PMID- 3053256 TI - Serum progesterone in the diagnosis of ectopic pregnancy: a valuable diagnostic test? AB - The value of a single serum progesterone (P) assay in the diagnostic work-up of suspected ectopic pregnancy was investigated in 89 patients with ectopic pregnancy and 27 patients with incomplete abortion. Reference values for P in the blood were obtained from 77 patients with normal intrauterine pregnancies in the first trimester. With the use of a discriminatory level of 63 nmol/l (20 ng/ml), sensitivity of 92%, specificity of 84%, positive predictive index of 90%, and negative predictive index of 87% were achieved. Addition of pelvic ultrasound further improved the diagnostic accuracy. It is concluded that serum P measurement offers a valuable adjunct to existing methods of diagnosis of ectopic pregnancy. PMID- 3053257 TI - [Thymostimulin induction of the synthesis of a factor intensifying the bactericidal activity of macrophages]. PMID- 3053258 TI - [Effect of destruction of the lateral septal nucleus on the sensitivity of the testes to chorionic gonadotropin]. PMID- 3053259 TI - [Effect of proteolytic enzymes on the functional activity of the neuron]. AB - Different neuronal responses to electrical stimuli under the effect of proteolytic enzymes, were shown. Different mechanisms of neuronal responses to stimuli were shown to depend on disturbances in neuroglial interrelationships. Two forms of the firing rate transformation depending on the type of stimulation, were described. PMID- 3053260 TI - [Interrelation of the hormonal indices and the cytologic characteristics of the germ cells]. AB - The analysis of the maturation of follicular oocytes as well as of the state of gametes after induction of ovulation with PMSG or with GnRH in intact and androgenized rats revealed the dependence of the heterogeneity extent (in morphologic and chromosomal damages) of population of germ cells maturing in vivo and in vitro upon the character of gonadotropin regulation disturbances of folliculogenesis. Stable disorder of reproductive function (androgenization) is followed by a significant increase in number of abnormal gametes under its hormonal correction. PMID- 3053261 TI - [Petr Stepanovich Kupalov (on the centenary of his birth)]. PMID- 3053262 TI - Quandaries in reproductive technology. PMID- 3053263 TI - The psychodynamics of dental anxiety and dental phobia. AB - This article deals with a broad overview of the incidence and treatment of dental anxiety and dental phobia. The dentist's position in this phenomenon is explored, and the basic psychological principles and modalities are discussed. This article serves to prepare the reader for the material presented in the ensuing articles. PMID- 3053264 TI - The challenge of fearful and phobic children. AB - Many children are fearful of going to the dentist. Professionals have the opportunity to avoid causing long-term anxiety disorders by providing a safe environment and offering children opportunities to overcome their normal childhood fears during early visits. It is more important to focus on a positive interaction between dentist and child rather than the completion of the dental procedure. PMID- 3053266 TI - Behavioral treatments for adult dental avoidance. A stepped-care approach. AB - The growing research literature dealing with the psychologic treatment of dental fear and avoidance suggests several interventions as effective, but provides little guidance in choosing among them. Under these circumstances, experienced psychologic practitioners may choose among these interventions on the basis of their own clinical impressions as to which treatment might be best suited to each patient they see. An alternative approach is the stepped-care approach in which the least expensive/most practical intervention is implemented, with more costly or complex procedures implemented only if the less expensive intervention proves unsuccessful. Each of the behavioral treatments for dental fear that has received researchers' attention is briefly described, and they are ordered in terms of their costs in professional time. Finally, an account of the special precautions needed to implement a stepped-care strategy for the reduction of dental fear is outlined. PMID- 3053265 TI - Pretreatment modeling. A technique for reducing children's fear in the dental operatory. AB - Research on modeling indicates that this technique offers dentists a means of reducing fear in child patients of all ages. As a preventive measure used with children who have had no prior exposure to dental treatment, it can be particularly efficacious. Based on the assumption that much of adult dental avoidance is based on dental fears acquired in childhood treatment, the reduction of children's dental fear would have a positive effect on the individual's tendency to seek out dental health care throughout his or her lifespan. For the dentist, there are also short- and long-term benefits. Dental management of the child is prerequisite to providing good dental care. Pedodontics as a specialty recognizes behavioral management of the child cannot be separated from the quality of the dentist's work. Fear has been identified as an important factor in disruptive behavior of school age children in the dental office. Practicing dentists consider the fearful, disruptive child to be among the most troublesome of problems in their clinical work. The child must cooperate or at least passively comply with the dentist's procedures in order to have the technical work completed. By reducing disruptive patient behavior (crying, screaming children whose peripheral and gross motor movements often make direct contact with the dentist or his equipment) the most unpalatable aspect of pediatric dentistry is minimized. Further, the actual time for treatment becomes shorter rather than longer. Although modeling is not restricted to videotape media, the emergence of current videotape technology provides the practitioner with the means for incorporating patient viewing of prerecorded modeling tapes as part of the usual waiting period. Such a procedure would mean that in the long run, the dentist will spend more time doing dentistry and less in behavioral management tasks. PMID- 3053267 TI - Biofeedback therapy in the treatment of dental anxiety and dental phobia. PMID- 3053268 TI - Hypnosis in the treatment of dental fear and phobia. AB - The term hypnosis is currently used to define an area of research and treatment that employs suggestion. Within this area, suggestion refers to the induction of expectancies by implicit or explicit means, usually involving concentration and the expectancy that the suggested results are possible. This use of suggestion differs from the common use of the term suggestion, which is a logical offering for a change in behavior or thought. The long history of hypnosis is testimony to its effectiveness, although there has been controversy as to why it works. Patient selection is important. Further, fear must be distinguished from phobia. Combined with other treatment techniques, such as systematic desensitization, it is a powerful behavior modification method. To prevent accidental delivery of suggestions that may be counterproductive to treatment, the study of hypnosis is important even to those health care professionals who have no intention of employing it in their practice. PMID- 3053269 TI - Adult patient-dentist relationship. AB - The author considers the adult patient dentist relationships from three vantage points: (1) the relationships of medical and dental practitioners; (2) the dynamic and anatomy of dental encounters; and (3) the essentials of clinical work, in which relationships are important. PMID- 3053271 TI - Patient satisfaction with dental care. AB - The feedback studies illustrate the fact that it is possible for the individual practitioner or member of a group practice to take positive steps to enhance satisfaction among his or her patients. Simply knowing about the general components of patient satisfaction is not enough; one needs to know about the views of one's own patients so that effective steps can be taken to improve them. The steps may include changes in office policies or procedures, facilities or staff changes, or even changes in the dentist's interpersonal approach. In any case, the evidence suggests that efforts to improve will be rewarded by more satisfied patients who will be more likely to stay as clients and, perhaps, more readily accept treatment and more frequently refer friends to the practice. The consistent role of dentists' interpersonal skills suggests that dental schools could contribute to patient satisfaction by providing more interpersonal skills development in their curricula. This was a major recommendation of the 1984 Future of Dentistry Final Report. The evidence presented in this chapter firmly supports that recommendation. PMID- 3053270 TI - Dental anxiety. Assessment, reduction and increasing patient satisfaction. AB - A conceptual schema relating a number of patient and dentist variables to patient anxiety reduction and satisfaction was presented. Evidence was examined that indicates that patient compliance with preventive and treatment regimens can be influenced in a major fashion through variables that reduce anxiety and increase satisfaction with the dentist. An accounting of several studies that investigated behavioral strategies for reducing patient stress during dental procedures indicated that recorded relaxation instructions and the active distraction provided by playing a video game can be effective anxiety-reducing treatments. Attempts to find other useful behavioral strategies were unsuccessful--only relaxation and distraction were consistently successful in reducing stress in moderately anxious patients. Observations of dentist-patient interaction in the context of the behavioral strategy studies indicated that the doctor-patient relationship was an important dimension associated with patient anxiety reduction and satisfaction. A series of investigations were conducted to elucidate the dentist behaviors that were associated with these variables. We found that the dentist behaviors most closely associated with patient satisfaction were those portraying empathy, friendliness, and a calm, competent image to the patient. The most important behavior associated with anxiety reduction was the dentist's explicit promise to prevent pain. Other dentist behaviors--friendliness, being calm, giving moral support--were seen as providing an appropriate behavioral context in support of the pledge to prevent pain. Finally, we considered the need for the dentist to be aware of patient anxiety in order to effectively deal with it. If nothing else, asking about anxiety gives the patient permission to express concerns that are present. If the patient is not anxious, asking about anxiety will not produce it. Two types of measuring instruments were considered in relation to assessing anxiety. The Dental Anxiety Scale, a brief four-item scale, was judged to be a reliable and valid method for assessing general dental anxiety. In addition, a new interval scale of anxiety response was discussed for those occasions on which it is important to assess the patient's response to treatment. PMID- 3053273 TI - Differentiating anxiety-panic disorders from psychologic dental anxiety. AB - It has always been believed that fear and anxiety of dental treatment was a simple continuum of experience that occurs in mild, moderate, or severe form. Past and present studies that attempt to both trace etiology and measure it reflect this view. The numerous studies that are concerned with methods of management are based on this accepted philosophy regarding the etiology of dental fear and anxiety. To a large extent, this may be true. However, there are some notable exceptions, and it is these cases that present the greatest management problem. Omitting the symptoms of fear and anxiety related to physical illness, drug withdrawal, or major mental illness, they present anxiety as a unidimensional learned problem usually conditioned by externally negative forces or experiences. They postulate that the fear and anxiety seen is due to a variety of factors. The interpretation of the definitions of fear, anxiety, and phobias by many in the profession that are presented in this issue also reflect the view that fear, anxiety, and phobias are learned or conditioned responses. This single minded view has determined much of our understanding and subsequent management of this problem in dentistry. PMID- 3053272 TI - Pharmacologic modalities in the management and treatment of dental anxiety. AB - This article defines the specific situations in which pharmacologic management and treatment of dental anxiety are appropriate and the services of a dentist anesthesiologist are required. PMID- 3053274 TI - A trial of oral 1 alpha-hydroxyvitamin D3 for ichthyosis. PMID- 3053275 TI - Resin-bonded bridges: 1. Methods and materials. PMID- 3053276 TI - Controlling active chronic periodontitis. PMID- 3053277 TI - Temporary pericoronal splinting: a new application of the acid-etch technique? PMID- 3053278 TI - What about tetanus? PMID- 3053279 TI - Resin-bonded bridges: 2. Clinical considerations. PMID- 3053280 TI - Rigid abdomen: an unusual cause. PMID- 3053281 TI - Mouse peritoneal macrophages produce CFU-gm and CFU-meg growth-promoting factors. AB - The effect of peritoneal macrophage-conditioned medium (macrophage CM) from lipopolysaccharide-injected mice on granulocyte-macrophage colony (CFU-gm) and megakaryocyte colony (CFU-meg) formation was examined using a plasma clot culture system. Macrophage CM stimulated mouse bone marrow cells to form CFU-gm and CFU meg in the presence of pokeweed mitogen-stimulated spleen cell-conditioned medium, but it had no effect on colony formation in the absence of exogenous colony-stimulating factors (CSF). CFU-gm and CFU-meg colony formation was enhanced by low concentrations of exogenous GM-CSF or Meg-CSF alone. These data demonstrate that macrophage CM contains an activity that sensitized CFU-gm and CFU-meg to exogenous CSF. PMID- 3053282 TI - Human trophoblast glycoproteins defined by monoclonal antibody 1D2. AB - A cell surface antigen has been defined by a monoclonal antibody 1D2, raised following immunisation with lectin-purified syncytiotrophoblast glycoproteins. 1D2 was nonreactive with any one of 8 common trophoblast proteins in immunodot. Analysis of nonreduced western blots of syncytiotrophoblast microvillous plasma membrane (StMPM) protein indicated that mAb 1D2 was reactive with a series of sialylated proteins with molecular weights of 16-22 kilodaltons. Immunoprecipitates of radiolabelled StMPM protein contained molecules that co migrated with placental alkaline phosphatase in addition to those identified by western blotting. This set of human trophoblast molecules has not been previously identified by monoclonal antibodies; the antigenicity is widely distributed in human tissues. PMID- 3053284 TI - The osteonectin family of proteins. PMID- 3053283 TI - Chemoprevention of cancer: phenolic antioxidants (BHT, BHA). AB - 1. The synthetic phenolic antioxidants (e.g. BHT, BHA) added to human and animal food are able to lengthen the life of organisms and lower the incidence of cancer caused by chemical compounds. 2. On the other hand they may not be rendered completely harmless since they can cause lung damage (BHT) or promote the action of some carcinogens (BHA). 3. They could act as compounds preventing cancer either via interception of harmful free radicals, activating the detoxifying enzymes of the body, inhibiting the formation of ultimately carcinogenic metabolites and their binding to DNA, and modifying the immune response of the organism. 4. Their action is influenced by their own chemical structure, the composition of carcinogen, the strain, sex and age of experimental animals, the tissue upon which they are supposed to act and the time of their administration in relation to the time of the carcinogen insult. 5. These compounds are concentrated in adipose tissue, liver and kidney. They are excreted within tens of hours mainly in urine. 6. The acceptable daily intake of BHA is at present considered to be 0.6 mg kg-1 body wt day-1. In spite of their possible tumor promoting properties they could not be considered overtly toxic. Their pronounced chemoprotective role against some forms of chemical carcinogenesis deserves considerable attention. PMID- 3053285 TI - Effects of insulin on pyruvate dehydrogenase in circulating lymphocytes from normal and diabetic rats. AB - 1. The in vivo and in vitro conditions which allow a response of rat circulating lymphocyte PDH to insulin are investigated. 2. In vivo tests show that inactive PDH (PDHi) prevails in diabetic rats and active PDH (PDHa) in hyperinsulinemic rats; in treated with insulin diabetic rats the PDHa/PDHi ratio (1.7) is similar to that of normal rats (PDHa/PDHi ratio = 2). 3. In vitro tests show a responsiveness of PDH to insulin only when 50 microM Ca2+ -Mg2+ and intact lymphocytes are used in the incubation medium. Insulin concentrations and contact time are important variables. PMID- 3053286 TI - The glycoconjugates of mammalian parasites with particular reference to Trypanosoma cruzi. PMID- 3053287 TI - Biogenesis of enzymes of peroxisomal beta-oxidation. PMID- 3053289 TI - Comparative enzymology of beta-oxidation. PMID- 3053288 TI - Acyl-CoA dehydrogenases, electron transfer flavoprotein and electron transfer flavoprotein dehydrogenase. PMID- 3053291 TI - Steroid receptors as oncogenes? PMID- 3053290 TI - Beta-oxidation of polyunsaturated fatty acids. PMID- 3053293 TI - Initial expression of type I procollagen in chick cardiac mesenchyme is dependent upon myocardial stimulation. AB - Formation of the atrioventricular (AV) mesenchyme is a critical step in early heart development. Endothelial cells are activated and transformed into a mesenchymal population that invades the cell-free myocardial basement membrane. This process can be duplicated in collagen gel culture, where it has been established that myocardium or its secretory products activate the endothelium. The purpose of the present study was to determine when these activated endothelial and/or mesenchymal cells start producing type I collagen in situ. These results were compared to those obtained from a culture model of mesenchyme formation. The production of type I collagen was monitored using a monoclonal antibody (M38) that recognizes the carboxy-terminal propeptide of human type I procollagen. The initial expression of the latter within activated AV endothelial and mesenchymal cells in ovo was 48 hr following activation. Prior to this time, only the myocardium was reactive with M38. AV explants of early hearts on collagen gels revealed staining of activated endothelial and mesenchymal cells with M38 after 48 hr in coculture with myocardial tissue. Explants that were prevented from activating (myocardium removed) never expressed the M38 antigen. Similarly, AV endothelial monolayers grown in the presence of myocardial conditioned medium activated and expressed type I collagen after 48 hr in culture, whereas those grown in standard medium did not. These results establish the initial expression of type I collagen within activated AV endothelium and mesenchyme. In addition, the data suggest that the expression of type I collagen within the AV mesenchyme may be dependent on extrinsic influences that induce the AV endothelium to transform into mesenchyme. PMID- 3053292 TI - Effect of estrogen on the expression of a cell-surface antigen associated with rat anterior pituitary somatotrophs. AB - The long-term in vivo effect of diethylstilbestrol (DES) on the expression of a cell-surface antigen associated with the anterior pituitary somatotroph was studied in two strains of female rats using double immunofluorescence techniques. Mab WHC-1, a recently generated and characterized monoclonal antibody, was used to detect the antigen associated with somatotrophs, whereas rabbit anti-rat prolactin (rPRL) and anti-human growth hormone (hGH) antisera were used to identify mammotrophs and somatotrophs, respectively. In F344 rats, Mab WHC-1 positive cells increased from 13.8 +/- 0.5% of total pituitary cells in normal anterior pituitaries to 34.2 +/- 4.0% in DES-induced pituitary tumors. The number of mammotrophs also increased significantly from 58.0 +/- 3.2% in controls to 75.9 +/- 2.2% in tumors. On the other hand, somatotrophs decreased significantly in number following ovariectomy (OVX) and DES implantation (19.7 +/- 0.5% vs. 6.1 +/- 1.2%). Based on double immunofluorescence, the percentage of Mab WHC-1 positive cells, which were somatotrophs, decreased from 85.5 +/- 2.7% in normal controls to 6.7 +/- 1.5% in DES-induced tumors. On the other hand, the percentage of Mab WHC-1-positive cells which were mammotrophs increased significantly from 14.0 +/- 1.4% to 86.1 +/- 1.8% following OVX and DES implantation. A similar change was found in the number of somatotrophs and mammotrophs following the same treatment in Sprague-Dawley (SD) rats which did not develop pituitary tumors. In contrast to F344 rats, the number of Mab WHC-1-positive cells in SD rats decreased significantly from 32.4 +/- 2.8% in sham-operated controls to 19.3 +/- 2.9% in OVX + DES-implanted rats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3053295 TI - Insulin-related molecules and insulin effects in the sea urchin embryo. AB - Insulin, the polypeptide hormone secreted by the differentiated pancreas, may play a role in vertebrate development at prepancreatic stages. In an invertebrate embryo, the sea urchin Strongylocentrotus purpuratus, we now find that insulin modulates the levels of developmentally regulated mRNAs of different lineages (one ectoderm-specific, one mesoderm-specific, and one found in all cell types). Using indirect immunofluorescence, we have localized a molecule which shares antigenic determinants with mammalian insulin in the unfertilized egg as well as in the gut of pluteus larva sea urchins. In addition, Southern hybridization reveals high similarity between sea urchin DNA sequences and the human insulin receptor gene. Our results suggest the presence of an insulin/insulin receptor related system in sea urchin development. PMID- 3053294 TI - Migratory behavior of cells on embryonic retina basal lamina. AB - In order to study cell translocation in vitro on a physiological substrate a novel cell migration assay was developed using the inner limiting membrane of the avian embryonic retina. The matrix sheet consists of a laminin-rich basal lamina covered by a dense layer of neuroepithelial endfeet. The retina basal lamina does not contain fibronectin. Cells translocating on this substrate displace the neuroepithelial endfeet, leaving behind tracks in the endfeet monolayer. Motility of cells and the relative forward to lateral migration can be quantitated by measuring lengths, widths, and areas of the tracks. Using this assay system, the conditions and patterns of cell migration for a variety of cells have been examined. In the absence of serum all cell types show only minor migratory activity and addition of serum to the culture medium always enhances the rate of cell migration in a saturable, dose-response manner. The serum cannot be replaced by fibronectin or vitronectin (serum spreading factor). For maximum cell migration, serum has to be constantly present in the medium; however, 58% cell migration is obtained in serum-free medium when the matrix is preincubated with serum. According to the area and linearity of the tracks, the migratory behavior of the different cells can be classified into three groups: (i) fibroblasts and the nonpigmented Bowes melanoma cells form straight and long tracks; (ii) glioma, sarcoma, and carcinoma cells from straight but short tracks, and (iii) neuronal tumor cells, epithelial cells, and pigmented B16 melanoma cells form wide and short tracks. Comparative studies with low and high metastatic clones of tumorgenic cell lines show that migratory activity and metastatic potential of cells do not necessarily correlate. Finally, we show that fibroblasts deposit fibronectin fibrils on their paths as they migrate on the basal lamina. Fibronectin trails are also seen when fibroblasts are cultured on plain basal laminae that are pretreated with detergent to remove the endfeet monolayer. Likewise, when fibroblasts are cultured in the presence of antifibronectin antibodies, the fibronectin secreted by cells is detectable. Due to antibody treatment the cellular fibronectin is precipitated and its normal fibril formation is inhibited; however, the translocation of fibroblasts is not impaired. PMID- 3053296 TI - Localization of the expression of types I, III, and IV collagen, TGF-beta 1 and c fos genes in developing human calvarial bones. AB - Total RNA extracted from developing calvarial bones of 15- to 18-week human fetuses was studied by Northern hybridization: in addition to high levels of type I collagen mRNAs, the presence of mRNAs for type III and type IV collagen, TGF beta and c-fos was observed. In situ hybridization of sections containing calvarial bone, overlying connective tissues, and skin was employed to identify the cells containing these mRNAs. Considerable variation was observed in the distribution of pro alpha 1(I) collagen mRNA in osteoblasts: the amount of the mRNA in cells at or near the upper surface of calvarial bone was distinctly greater than that in cells at the lower surface, indicating the direction of bone growth. High levels of type I collagen mRNAs were also detected in fibroblasts of periosteum, dura mater, and skin. Type III collagen mRNA revealed a considerably different distribution: the highest levels were detected in upper dermis, lower levels were seen in fibroblasts of the periosteum and the fibrous mesenchyme between bone spiculas, and none was seen in osteoblasts. Type IV collagen mRNAs were only observed in the endothelial cells of blood capillaries. Immunohistochemical localization of type III and IV collagens agreed well with these observations. The distribution of TGF-beta mRNA resembled that of type I collagen mRNA. In addition, high levels of TGF-beta mRNA were observed in osteoclasts of the calvarial bone. These cells, responsible for bone resorption, were also found to contain high levels of c-fos mRNA. Production of TGF-beta by osteoclasts and its activation by the acidic environment could form a link between bone resorption and new matrix formation. PMID- 3053297 TI - Developmental modulation of a glial cell-associated glycoprotein, 5B12, in an insect, Acheta domesticus. AB - The expression of an insect (Acheta domesticus) adult glial cell-specific antigen, 5B12 undergoes major changes during development. The 5B12 antigen is detected as early as 20-25% of embryonic development, when immunoreactivity is distributed throughout the periphery, present at the luminal surface of epithelial cells which compose developing limb buds, sensory appendages, and the body cavity. The antigen is also localized on the cell surface of neural elements within commissural tracts in the embryonic CNS. 5B12 is secreted extracellularly in the periphery, where it is associated with the embryonic basal lamina in developing cercal sensory appendages. Luminal surface expression is transient, and disappears by 95% of embryonic development. As development proceeds, 5B12 distribution becomes more restricted, so that in the adult the antigen is predominantly associated with specific glial elements within the nervous system where it occurs as a specialized component of the extracellular matrix. The 5B12 antigen is also associated with discrete central and peripheral fiber tracts. Antigen 5B12 is present in whole embryos and in the adult CNS as a Mr 185-kDa glycoprotein. Distinct carbohydrate moieties with chondroitin sulfate-like properties are situated on the 5B12 epitope. Thus the glia-associated 5B12 macromolecule has the characteristics of a small proteoglycan. Based upon features of its distribution, pattern of spatiotemporal expression, and biochemical properties, it is speculated that 5B12 participates in events related sequentially to the development and the function of the insect nervous system. PMID- 3053299 TI - Acetylated alpha-tubulin in microtubules during mouse fertilization and early development. AB - alpha-Tubulin in the microtubules of mouse oocytes and embryos is acetylated in a specific spatial and temporal sequence. In the unfertilized oocyte, a monoclonal antibody to the acetylated form of alpha-tubulin is bound predominantly at the poles of the arrested metaphase meiotic spindle. The labeling intensity of the spindle microtubules is weaker as observed by immunofluorescence using oocytes double-labeled for total tubulin and acetylated alpha-tubulin, and as measured by immuno high-voltage electron microscopy (immunoHVEM) with colloidal gold; cytasters are not acetylated. At meiotic anaphase, the spindle becomes labeled, and by telophase and during second polar body formation only the meiotic midbody is acetylated. The sperm axoneme retains its acetylation after incorporation though the interphase microtubules are not detected. First mitosis follows a pattern similar to that observed at the second meiosis and during interphase only the mitotic midbodies are acetylated. After treatment with cold, colcemid, or griseofulvin, the remaining stable microtubules are acetylated, but immunoHVEM observations suggest that these fibers might not have been acetylated prior to microtubule disruption. Taxol stabilization does not alter acetylation patterns. Acetylated microtubules are not necessarily old microtubules since acetylated fibers are observed at 30 sec after cold recovery. These results show the presence of acetylated microtubules during meiosis and mitosis and demonstrate a cell-cycle-specific pattern of acetylation, with acetylated microtubules found at the centrosomes at metaphase, an increase in spindle labeling at anaphase, and the selective deacetylation of all but midbody microtubules at telophase. PMID- 3053300 TI - President's address. PMID- 3053298 TI - Sea urchin primary mesenchyme cells: relation of cell polarity to the epithelial mesenchymal transformation. AB - In euechinoid sea urchin embryos, a subset of epithelial cells in the wall of the blastula become pulsatile, elongate, lose connections with their neighboring cells, and move into the blastocoel to form the primary mesenchyme cells. The Golgi apparatus and microtubule organizing center (MTOC) are located at the apical end of these epithelial cells. We show that as primary mesenchyme cells begin to move into the blastocoel, the Golgi apparatus and MTOC move to a new position adjacent to the apical side of the nucleus. They do not move to a position between the nucleus and the leading (i.e., basal) end of the cell as they do in cultured fibroblasts undergoing directed migration. In addition, we have inhibited the movement of membranous vesicles to the cell surface by incubating embryos in the ionophore monensin. We have used antibodies to msp130, a primary mesenchyme cell surface-specific glycoprotein, to demonstrate that monensin inhibits the movement of msp130-containing vesicles to the cell surface. Despite the inhibition of membrane shuttling by monensin, primary mesenchyme cells ingress on schedule and display normal cell-shape changes. We draw two conclusions from our data. First, the cellular elongation that characterizes ingression is not due to the local insertion of membrane at the leading (basal) end of the cell. Second, ingression does not depend upon establishment of the same cell polarity required for fibroblasts to carry out directed cell migration. PMID- 3053301 TI - Biosynthetic regulation of endogenous hamster insulin and exogenous rat insulin II in transfected HIT cells. AB - To investigate mechanisms underlying biosynthetic regulation of an insulin gene, the rat insulin II gene was introduced into hamster beta-cells (HIT) by cotransfection with the neomycin phosphotransferase-selectable marker. The insulin gene fragment was 2.2 kilobases (kb) in length and contained all exons, introns, and approximately 700 base pairs (bp) of 5'-flanking DNA and 300 bp of 3'-flanking DNA. The HIT cell was known to have endogenous hamster insulin production under regulation by glucose and dexamethasone. In a pool of stably transfected cells (HIT M62pR2), rat insulin II and hamster insulin were produced at comparable rates. Glucose (20 mM) stimulated cellular [3H]leucine labeling of both hamster insulin and rat insulin II by approximately twofold. Addition of 10( 6) M dexamethasone to media containing 11.1 mM glucose inhibited biosynthesis of both hamster insulin and rat insulin II by greater than 90%. Thus, with both positive and negative biosynthetic regulation, changes in the cellular labeling of exogenous rat insulin II were qualitatively and quantitatively similar to those of the endogenous hamster insulin. These data suggest that the 2.2-kb rat insulin II gene fragment contained sufficient information for both expression and apparently "normal" biosynthetic regulation of exogenous rat insulin II (when compared with endogenous hamster insulin) in response to glucose and dexamethasone. PMID- 3053302 TI - Stimulation of glucose production through hormone secretion and other mechanisms during insulin-induced hypoglycemia. AB - To assess the role of counterregulatory hormones per se in the response to continuous insulin infusion, overnight-fasted dogs were given 5 mU.kg-1.min-1 insulin intraportally either alone (INS, n = 5), with glucose to maintain euglycemia (INS + GLU, n = 5), or with glucose and hormone replacement [i.e., glucagon, epinephrine, norepinephrine, and cortisol infusions (INS + GLU + HR, n = 6)]. The increases in counterregulatory hormones that occurred during insulin induced hypoglycemia were simulated in the latter group. In this way, it was possible to separate the effects of hypoglycemia per se from those due to the associated counterregulatory hormone response. Glycogenolysis and gluconeogenesis were measured with a combination of tracer ([ 3-3H]glucose and [U-14C]alanine) and hepatic arteriovenous (AV) difference techniques during a 40-min control and a 180-min experimental period. Insulin levels increased similarly in all groups (to congruent to 250 microU/ml), whereas plasma glucose levels decreased in INS (115 +/- 3 to 41 +/- 3 mg/dl; P less than .05) and rose slightly in both INS + GLU (108 +/- 2 to 115 +/- 4 mg/dl; P less than .05) and INS + GLU + HR (111 +/- 3 to 120 +/- 3 mg/dl; P less than .05) due to glucose infusion. Glucagon, epinephrine, norepinephrine, and cortisol were replaced in INS + GLU + HR so that the increments in their levels were 102 +/- 6, 106 +/- 14, 117 +/- 9, and 124 +/- 37%, respectively, of their increments in INS. At no time was there a significant difference between the hormone levels in INS and INS + GLU + HR. The rise in the counterregulatory hormones per se accounted for only half (53 +/- 9% by the AV difference method and 54 +/- 10% by tracer method) of the glucose production associated with hypoglycemia resulting from insulin infusion. The rate and efficiency of alanine conversion to glucose in the hormone-replacement studies were only 29 +/- 10 and 50 +/- 27% of what occurred during hypoglycemia induced by insulin infusion. In conclusion, the counterregulatory hormones alone (i.e., without accompanying hypoglycemia) can account for only 50% of the glucose production that is present during insulin-induced hypoglycemia. The remaining 50%, therefore, must result from effects of hypoglycemia other than its ability to trigger hormone release. PMID- 3053304 TI - Regulation of glucose-transporter gene expression by insulin in cultured human fibroblasts. AB - To clarify the effect of insulin on glucose-transporter (GT) biosynthesis, we determined GT mRNA levels in human cultured skin fibroblasts, using HepG2 GT cDNA as a probe. Insulin specifically increased the GT mRNA level in a time- and dose dependent manner. Time-course study demonstrated that the mRNA level peaked within 3 h of insulin (1 x 10(-7) M) addition. After remaining elevated for several hours, mRNA decreased and returned to the basal level after 24 h. In the cell strains from seven normal subjects, the mean (+/- SE) GT mRNA level determined after 3 h of treatment with 1 x 10(-7) M insulin was 164.3 +/- 8.5% of the level found in untreated control cells. The insulin dose-response curve of GT mRNA levels showed that the maximum stimulation was elicited at 1 x 10(-7) M, and the half-maximum stimulation occurred at approximately 5 x 10(-10) M. Degradation rates of GT mRNA determined in the presence of actinomycin D were not different between insulin-treated and untreated cells. These results suggest that insulin increases GT gene expression in cultured human fibroblasts. PMID- 3053303 TI - Characterization of new oral antidiabetic agent CS-045. Studies in KK and ob/ob mice and Zucker fatty rats. AB - CS-045 is a new oral antidiabetic agent that was effective in insulin-resistant diabetic animal models, including the KK mouse, the ob/ob mouse, and the Zucker fatty rat. CS-045 was not effective in the streptozocin-treated mouse, an insulin deficient diabetic animal model. In fed KK mice, CS-045 lowered the plasma glucose levels in a dose-dependent manner after a single oral administration, and the hypoglycemic effect lasted for at least 18 h. In normal rats, however, plasma glucose levels were not changed after administration of CS-045. CS-045 when given chronically (2 wk) to diabetic KK and ob/ob mice as a 0.2% food admixture dramatically improved hyperglycemia, hyperinsulinemia, and hypertriglyceridemia to near-normal values and decreased plasma lactate, free fatty acid, and ketone body levels without reducing food intake or body weight. In the obese Zucker fatty rat, oral administration of CS-045 had a similar effect in lowering plasma glucose, insulin, triglyceride, free fatty acid, lactate, and ketone body levels. The CS-045-treated Zucker fatty rats showed increased glucose tolerance and decreased insulin secretion in response to oral glucose. After 9 days of treatment, insulin binding to adipocyte plasma membranes from both CS-045-treated Zucker fatty rats and KK mice was increased. Furthermore, 2-deoxyglucose uptake in CS-045-treated adipocytes was increased and the insulin dose-response curve was shifted to the left. These findings suggest that CS-045 increases not only insulin sensitivity but also insulin responsiveness. Based on its pharmacological profile, CS-045 is a new orally effective antidiabetic agent that may reduce abnormalities of glucose and lipid metabolism in obese and non-insulin-dependent diabetes mellitus patients with insulin resistance. PMID- 3053305 TI - Autoantibodies in nonobese diabetic mice immunoprecipitate 64,000-Mr islet antigen. AB - In insulin-dependent diabetes mellitus (IDDM) in humans and BB rats, islet cell autoimmunities associated with autoantibodies to a beta-cell protein of 64,000 Mr (64K) have been described. We report that sera from newly diagnosed nonobese diabetic (NOD) mice similarly contain an autoantibody that immunoprecipitates 64K autoantigen from detergent lysates of [35S]methionine-labeled murine islet cells. The autoantibody was detectable by weaning; it disappeared within weeks after diabetes onset and was absent in older nondiabetic NOD mice as well as all of three non-diabetes-prone control strains tested. The 64K beta-cell autoantigen may be a critical target in the immunopathogenesis of IDDM. PMID- 3053306 TI - Control of myogenesis in the mouse myogenic C2 cell line by medium composition and by insulin: characterization of permissive and inducible C2 myoblasts. AB - Using subcloning and manipulations of culture conditions we have isolated from the mouse myogenic cell line C2 a variant cell line that we named inducible. Unlike the progenitor cells that are referred to as permissive, inducible myoblasts differentiate poorly in Dulbecco modified Eagle medium plus fetal calf serum (FCS) and require the presence of insulin at a high concentration (1.6 10( 6) M) or insulin-like growth factor I (IGFI) at a lower concentration (2.5 10(-8) M) to differentiate. Permissive and inducible myoblasts fail to differentiate when grown in MCDB202 medium plus 20% FCS, even after a prolonged arrest in G1 phase. This shows that an arrest in G1 is in itself insufficient to trigger terminal differentiation. Both cell types also exhibit distinct patterns of accumulation of muscle mRNAs corresponding to sarcomeric actins and myosin light chain MLC1A. The possibility that these two cell lines might represent two different stages of the progression of myoblasts toward terminal differentiation is discussed. PMID- 3053307 TI - The chemistry and biology of unusual DNA structures adopted by oligopurine.oligopyrimidine sequences. AB - A family of unusual DNA structures has been discovered in segments with predominantly purines in one strand (pur.pyr sequences). These sequences are overrepresented in eukaryotic DNA and have been mapped near genes and recombination hot spots. When cloned into recombinant plasmids, many pur.pyr sequences are reactive to chemical and enzymic probes that are generally specific for single-stranded DNA. An intramolecular triplex is adopted by mirror repeats of G's and A's. Other non-B DNA structures adopted by similar sequences remain to be fully clarified but may be a family of related conformations. It is likely that these unorthodox structures play an important role in the function of the eukaryotic genome. PMID- 3053308 TI - Collagen autoimmunity and arthritis. AB - Collagen-induced arthritis in animals is an example of polyarthritis that sufficiently resembles human rheumatoid arthritis to be used as a model. It is caused by immunizing susceptible animals with type II collagen isolated from articular cartilage. Susceptibility is genetically determined and linked to the major histocompatibility locus. It is important because some human arthritis is also associated with major histocompatibility genes and may be caused or aggravated by the presence of autoimmunity to normal cartilage components. Collagen-induced arthritis is also important because it is an example of immunologically mediated joint destruction, which may share some of the mechanisms present in human disease. Although it is caused by autoimmunity to collagen, susceptibility and responsiveness to type II collagen are not completely correlated, and there are examples of animals with high levels of collagen immunity who do not develop arthritis. The initial lesion appears to be the deposition of an antibody on the surface of articular cartilage, which precedes development of overt arthritis by several days. Disease can be readily transferred with specific antibody. Arthritogenic antibodies appear to have restricted epitope specificity, which may partially explain the disparities between responsiveness to immunization with collagen and susceptibility to arthritis, but precise delineation of the epitopes involved has not yet been accomplished. Complement activation also appears to be intimately involved since the disease correlates with the presence of high levels of complement-binding IgG isotypes, and passive transfer is possible only into complement-sufficient recipients. Inflammation progresses rapidly so that cartilage destruction and marginal erosion develop over a period of a few days. Collagen-induced arthritis offers a unique opportunity to study autoimmune-mediated arthritis in which the inducing antigen is well characterized and readily available. Analysis of the disease has permitted the proposal of a schema for its pathogenesis. PMID- 3053309 TI - Nuclear cardiology. PMID- 3053311 TI - Presentation of the Friedenwald medal to Frank Pickering Brooks, M.D. PMID- 3053310 TI - Digital subtraction angiography of right cervical aortic arch. AB - This report describes a 7 year old boy with a history of recurrent respiratory infections, a 3/6 ejection systolic murmur and a right supraclavicular systolic thrill. Chest x-ray showed widening of the superior mediastinum towards the right, and barium esophagram demonstrated anterior displacement and compression of the esophagus. Cross-sectional echocardiography revealed a transverse aortic arch segment. Thus, the findings described above suggested the noninvasive diagnosis of the right cervical aortic arch. A peripheral intravenous digital subtraction angiography was necessary to evaluate the origin of the epi-aortic arteries and it proved adequate for the follow-up of these patients. PMID- 3053312 TI - Low-dose antacids or cimetidine for duodenal ulcer? AB - In a double-blind, randomized, multicenter trial 150 consecutive outpatients with endoscopically verified duodenal ulcer were treated with either a low-dose antacid regimen (1 tablet q.i.d.; acid-neutralizing capacity, 120 mmol/day), or cimetidine (800 mg nocte). After 4 wk of treatment control gastroscopy showed ulcer healing in 54 of 76 patients (71.1%) in the antacid group, as compared with 58 of 74 patients (78.4%) in the cimetidine-treated group. The difference in healing rate of 7.3% (95% confidence interval, -6.5% to +21.1%) was not statistically significant. The symptomatic effect, measured as number of days and nights with ulcer pain, was also quite similar in the two treatment groups. However, the number of days with pain was significantly lower in the first week of treatment in the antacid group (p less than 0.01). Thus, the efficacy of a low dose antacid tablet regimen approximated that of cimetidine (800 mg nocte) in the treatment of duodenal ulcer patients. PMID- 3053313 TI - Effect of starch malabsorption on colonic function and metabolism in humans. AB - To study the impact of starch on colonic function and metabolism, 12 healthy volunteers consumed a controlled diet rich in starch for two 4-wk periods. In one of the study periods they received the glucosidase inhibitor acarbose (BAY g 5421) and placebo in the other. Stool wet weight increased by 68%, stool dry weight by 57%, fecal water content by 73%, and the mean transit time by 30% on acarbose. Breath hydrogen was significantly higher on acarbose, indicating stimulated carbohydrate fermentation in the colon. Fecal bacterial mass (+78%), total stool nitrogen (+53%), bacterial nitrogen (+200%), and stool fat (+56%) were higher in the acarbose than in the control period. The stimulation of fermentation in the human large intestine may be important in colonic and possibly other diseases. PMID- 3053314 TI - No effect of oral testosterone treatment on sexual dysfunction in alcoholic cirrhotic men. AB - The prevalence and course of sexual dysfunction was evaluated in 221 alcoholic cirrhotic men participating in a double-blind, placebo-controlled study on the effect of oral testosterone treatment on liver disease. At entry, 67% (95% confidence limits, 61%-74%) complained of sexual dysfunction. Sexual dysfunction was significantly (p less than 0.05) associated with lower serum concentrations of testosterone, non-protein-bound testosterone, and non-sex hormone-binding globulin-bound testosterone. The significant associations between sexual dysfunction and non-protein-bound and non-sex hormone-binding globulin-bound testosterone concentrations disappeared, however, when age, ethanol consumption, and severity of liver disease were included as covariates in the analysis. During follow-up (median 30 mo, range 1-48 mo) sexual dysfunction improved significantly (p less than 0.05) at 6, 12, and 24 mo. Furthermore, the reported libido and erectile and ejaculatory function improved significantly at the end of the follow up period (p less than 0.01). However, the testosterone-treated patients did not differ significantly from the placebo-treated patients regarding any of the changes in sexual function. In conclusion, oral testosterone treatment does not significantly influence the type or course of sexual dysfunction in alcoholic cirrhotic men. However, sexual function improved after reduction of ethanol consumption in these patients. PMID- 3053315 TI - Effects of synthetic human gastric inhibitory polypeptide on splanchnic circulation in dogs. AB - Changes in blood flow in the celiac artery, superior mesenteric artery, and pancreas in response to an intravenous injection of synthetic human gastric inhibitory polypeptide (GIP) were determined simultaneously and continuously in anesthetized dogs, using a transit-time ultrasonic flowmeter and a laser-Doppler flowmeter. Injection of GIP significantly increased superior mesenteric arterial flow in a dose-related manner (by 9%, 43%, and 139% at 30 s after an injection at the doses of 3, 50, and 800 pmol/kg, respectively). In contrast, celiac arterial flow was not significantly altered by GIP at any of the three doses. Calculated vascular resistance in the superior mesenteric artery decreased after GIP infusion, whereas that in the celiac artery was not changed by GIP. Pancreatic blood flow decreased significantly after GIP injection at the doses of 50 and 800 pmol/kg (by 11% and 17%, respectively). Our data indicate that there is a substantial difference in the hemodynamic responses to GIP among splanchnic organs, and suggest that GIP acts specifically on the mesenteric vasculature. PMID- 3053317 TI - Ultrasonography in chronic liver disease. PMID- 3053316 TI - Aortic stenosis, idiopathic gastrointestinal bleeding, and angiodysplasia: is there an association? A methodologic critique of the literature. AB - To assess the reported association between colonic angiodysplasia and aortic stenosis, we performed a quantitative and methodologic analysis of the literature. In four controlled studies that support an association between aortic stenosis and idiopathic gastrointestinal bleeding there are major methodologic deficiencies including the following: nonblinded data collection, noncomparable diagnostic examination, nonblinded ascertainment of exposure, and noncomparable demographic susceptibility. None of the studies directly assesses angiodysplasia. Additional case reports about aortic valve replacement used to treat bleeding from angiodysplasia are limited in number and in duration of follow-up. We conclude that the existing literature does not demonstrate an association between aortic stenosis and angiodysplasia. Further controlled evaluation of this topic would be useful. PMID- 3053318 TI - Presentation of the 1988 Rudolf Schindler Award to Bernard M. Schuman. PMID- 3053319 TI - Rudolf Schindler, MD: living with a Renaissance man. PMID- 3053321 TI - Colonoscopic decompression for acute colonic pseudo-obstruction (Ogilvie's syndrome) in transplant recipients. PMID- 3053320 TI - Absolute alcohol in esophageal vein sclerosis. AB - Absolute alcohol is a potentially optimal agent for sclerotherapy of esophageal varices. It is cheap and readily available. We compared the efficacy and safety of alcohol with those of a commonly used sclerosing agent, polidocanol. The study was planned to include patients with previous bleeding from esophageal varices randomly assigned to one of the two treatments. After the inclusion of the first 11 patients (6 in the polidocanol group and 5 in the alcohol group), however, the trial was interrupted because of serious complications in patients treated with alcohol (four major bleeding episodes and one esophageal stenosis). The two agents were of comparable efficacy in the small sample of patients studied. The complications were related to the presence of iatrogenic esophageal ulcers which were more frequent (100% vs. 30%) and significantly larger (mean, 1.4 cm vs. 0.7 cm, p less than 0.05) in patients treated with alcohol. PMID- 3053322 TI - The team approach to biliary tract intervention: current status of combined percutaneous-endoscopic techniques. PMID- 3053323 TI - [Transvaginal, ultrasound-controlled follicle puncture]. AB - Between December 1986 and November 1987, 588 transvaginal sonographically-guided oocyte retrievals, using a vaginal transducer, were performed for in vitro fertilization at the University of Bonn. All follicles were accessible. No complication occurred except one case of pelviperitonitis in a patient with preoperatively diagnosed sactosalpinx. Because this technique is safe, non invasive, and performable without general anaesthesia, we also retrieved eggs in patients despite poor hormonal values and endogenous LH-surge. In all, clinical pregnancies were diagnosed in 85 patients. Pregnancy rates per embryo transfer, oocyte retrieval and stimulated cycle, respectively were 19.0%, 14.5% and 12.0%. The advantages of the transvaginal technique, using a vaginal transducer compared with the laparoscopically-guided as well as with other sonographically-guided techniques of oocyte retrieval, are obvious (possibly without general anaesthesia, in general all follicles are accessible, very low complication rates, low discomfort to patient, and less amount of time and staff). Therefore, this technique seems to be the method of first choice. A laparoscopically-guided oocyte retrieval is only indicated, when a laparoscopic screening of pelvic organs or a gamete intrafallopian transfer are to be performed simultaneously. PMID- 3053325 TI - [Simultaneous intra- and extrauterine pregnancy]. AB - A report on a case of simultaneous intrauterine and extrauterine pregnancy associated with right-sided tubal pregnancy and rupture in the eighth week of pregnancy and spontaneous parturition in the 42nd week. The incidence rate and aetiological factors, as well as diagnostic possibilities and differential diagnostic considerations are presented. PMID- 3053324 TI - [In vitro fertilization: a comparison with results of the hamster ova penetration test (HOPT)]. AB - The hamster ova penetration test (HOPT) was performed in 82 patients simultaneously with in vitro fertilization of homologous oocytes (IVF). A penetration rate of greater than 20% of inseminated ova was considered to represent a positive test result. The results of HOPT correlated completely with the IVF in all couples with male factor sterility. In couples with female sterility only and in couples with a combination of female and male factors HOPT was falsely negative in 22 and 20% respectively. Despite a negative HOPT in vitro fertilization with formation of two pronuclei 12 to 18 hours after insemination occurred in these patients. Obviously, the interpretation of HOPT is dependent upon the definition of the cut-off point (in our case 20%). Our results support the opinion that there should be no exclusion from an IVF programme based only on a negative HOPT. It appears that any penetration rate greater than 0% should be considered as a sign of potential fertility. PMID- 3053326 TI - [Modern fertility technologies: challenges for psychosomatic anthropology]. AB - In the present paper the topic is considered on the basis of homologous in-vitro fertilization, taking homologous test-tube fertilization as representative of other fertilization methods. The first part of the article considers a number of obvious forms of damage and symptoms, the second the simplified anthropology on which medical fertilization methods are based and the challenge--arising from this--to develop a new psychosomatic anthropology, crystallized in the paradigm of the advent of a child. PMID- 3053327 TI - Isolation and structural characterization of insulin from the holocephalan fish, Chimaera monstrosa (rabbit fish). AB - Insulin has been isolated from the pancreas of the holocephalan fish, Chimaera monstrosa (rabbit fish), and characterized by automated Edman degradation and fast atom bombardment mass spectrometry. The primary structure of rabbit fish insulin was identical to that of insulin from the holocephalan fish, Hydrolagus colliei (Pacific ratfish), and contained 21 residues in the A-chain and 38 residues in the B-chain. The amino acid compositions of both rabbit fish and ratfish insulins demonstrated a value consistently lower than that expected for the leucine content of the peptides. It is suggested, therefore, that the insulins were probably isolated as a mixture of the intact peptides and components lacking the C-terminal leucine residue in the B-chain. PMID- 3053328 TI - Immunocytochemical evidence for the colocalization of neurotensin/xenopsin- and gastrin/caerulein-immunoreactive substances in Xenopus laevis gastrointestinal tract. AB - Distribution and association of neurotensin (NT)- and xenopsin (XP)-like peptides were investigated using immunocytochemical techniques in the amphibian gut. Antisera against both groups of peptides showed an identical distribution pattern of NT- and XP-positive cells in Xenopus laevis gastrointestinal tract. Immunolabeling of consecutive semithin sections revealed the coexistence of NT- and XP-like substances within cells of the stomach and small intestine. Recent reports of the colocalization of XP-like material with gastrin in mammalian G cells led us to study the association of NT/XP-like peptides with members of the gastrin/cholecystokinin (CCK)/caerulein (G/C) family in amphibians. The data obtained from immunolabeling serial sections with NT/XP-specific and G/C-specific antisera show that in some intestine NT/XP- and G/C-like peptides do exist in the same cells. In the stomach, however, G/C-like material is confined to endocrine cells of the antral region, while NT/XP-like substances occur in distinct cells accumulating in cardial glands but absent in the pyloric glands. Our findings thus indicate that in amphibian gastrointestinal tract there is some association between the regulatory peptide families NT/XP and G/C, similar to mammals. The regional distribution of both hormone families, however, is different from that in mammals. PMID- 3053329 TI - [Killer systems of Saccharomyces cerevisiae yeasts]. AB - The killer systems of Saccharomyces cerevisiae are a peculiar group of cytoplasmic symbionts of primitive eukaryotes. The genetic material of these symbionts is double-stranded RNA. Their basic properties are linearity of genome, its fragmentation, resulting in two separately replicating major and minor segments, and the ability to control the synthesis of secretory proteins- mycocins which can kill the taxonomically related strains. Secretion of mycocins also confers immunity to their action. The strains containing killer symbionts are toxigenic and resistant to their own toxins, while those with no killer double-stranded RNA are sensitive to mycocins. The killer systems of Saccharomyces cerevisiae possess some properties relevant to viruses and evidently are evolved during the evolution of infectious viruses. Occurrence of such systems in monocellular eucaryotic organisms is an example of genome complication in the course of putting together the virus-like components. The peculiarities of replication and expression of killer systems and their utilization for the construction of vector molecules are discussed. PMID- 3053331 TI - [Mutagenesis on cloned yeast genes. The mutation of the yeast gene comprising the plasmid and chromosome]. AB - The cells of Saccharomyces cerevisiae were transformed by plasmid pYG-007 treated in vitro with o-methylhydroxylamine. The plasmid consists of a portion of the bacterial plasmid with genes of resistance to ampicillin, chloramphenicol and tetracycline, 2 mkm yeast DNA and yeast genes ADE2 and LEU2. The collection of mutants containing a mutant allele of ADE2 gene within the plasmid was obtained. Interallelic complementation and that induced by suppression were studied in these ade 2 mutants. It was shown that all these induced ade 2 mutations were base-pair substitutions. Using the mechanism of conversion we managed to transfer the plasmid ade 2 mutations into the chromosome. Three pairs of strains carrying similar mutation in plasmid and chromosome were created. Analysis of frequency of reversions induced by UV-light and hydroxylaminopurine in the mutant ade2 locus comprised in the plasmid and chromosome showed that the former induced reversions in plasmid alleles less effectively than the latter. PMID- 3053330 TI - [Selective systems for obtaining recessive ribosomal suppressors in saccharomycete yeasts]. AB - Recessive mutations only occurring in two genes (ribosomal suppressors sup1 and sup2) can be obtained using special selective system. We demonstrate that the absolute selectivity of the system is based on selection for simultaneous reversions to prototrophy in mutants requiring adenine and histidine in haploids marked by two different nonsense mutations--his7-1 (UAA) and ade1-14 (UGA, this being identified in the present study). In support to this conclusion, we developed an analogous system utilising his7-1 (UAA) and lys2-87 (UGA). The selectivity of the system is shown to be influenced both by the choice of nonsense alleles and by genotypic background. PMID- 3053333 TI - Medicare confuses geriatricians, too. PMID- 3053334 TI - Psychoactive drugs in the elderly: antidepressants. AB - Clinical depression in community-dwelling elderly has been estimated to be as high as 13%. Depression is frequently seen in the typical geriatric scenario of concurrent medical illness, and the suicide rate among the elderly is disproportionately higher than other age groups. Recognizing depression in the typical geriatric setting of multiple medical problems and differentiating it from other psychologic disorders is discussed in this review, as is an approach to assessing suicidal potential. An enlightened approach to drug therapy is presented, incorporating knowledge of previous treatment response and anticipating therapeutic benefits and side effects in the individual patient. An update on electroconvulsive therapy, particularly in severe geriatric depression, is included. PMID- 3053332 TI - [Genomic fingerprinting of microorganisms: its use as a hybridization probe of phage M13 DNA]. AB - Hypervariable nucleotide sequences detected by hybridization with the phage M13 DNA probe were found in the chromosomal DNAs of certain pathogenic microbial species. DNA fingerprinting, based on hybridization of M13-probe with hypervariable chromosomal DNA sequences, opens new approaches to epidemiological analysis, epidemiological prognosis, taxonomy, and other theoretical and applied fields of bacteriology. PMID- 3053335 TI - Gout: how presentation, diagnosis, and treatment differ in the elderly. AB - Although frequently a typical acute monoarticular arthritis, gout in the elderly is often a chronic, polyarticular disease, sometimes with minimal inflammation. It can be associated with osteoarthritis and diuretic therapy, can follow minor trauma or medical illness, and is seen commonly in women. The management of gout in the elderly requires special consideration of the expected course of the disease and of the risks of medical therapy. PMID- 3053336 TI - Sexual dysfunction in postmenopausal women: the role of medical management. AB - A general decline in postmenopausal sexual activity, compounded by various physiologic changes related to the menopause, can be the source of much anxiety for older women. Many may wish to remain sexually active, yet fail to see that their problem is, in part, a medical one which a concerned physician can treat. Strategies for physician intervention, clinical management, and general support are reviewed for one of older patients' more undertreated maladies. PMID- 3053337 TI - Werner's syndrome associated with malignancies: five case reports with a survey of case histories in Japan. AB - We present 5 cases of Werner's syndrome associated with malignancies and a survey of 26 cases in the Japanese literature. Though tumors of mesenchymal origin have been reported in Werner's syndrome, 14 of the 31 cases cited in this paper developed carcinomas. Carcinoma of the thyroid gland was relatively high in frequency. The significance of carcinomas in Werner's syndrome should be further investigated. PMID- 3053338 TI - [The contribution of Prof. E. Ts. Andreeva-Galanina to the study of occupational vibration and noise (on the centenary of her birth)]. PMID- 3053339 TI - [Industrial hygiene and toxicology problems in the manufacture of titanium and its compounds (a review of the literature)]. PMID- 3053340 TI - [Heart transplantation. The initial successful operations and discussion of the problem]. PMID- 3053341 TI - [Formation of an invagination esophageal-gastric (intestinal) anastomosis using Bloxhin's expanding suture]. PMID- 3053342 TI - [Extirpation of the esophagus without thoracotomy]. PMID- 3053343 TI - Primary squamous cell carcinoma of the endometrium. AB - A case of squamous cell carcinoma of the endometrium which fulfills Fluhmann's criteria is presented. This case is reviewed together with additional 25 cases which had been already reported in various papers in the literature. The pathogenesis and the etiologic factors associated with primary squamous cell carcinoma of the endometrium are discussed. A review of the literature indicates that this rare malignancy is found in older patients, is highly malignant and carries a poor prognosis as myometrial invasion, and local extension or metastasis is found in 80% of the patients. PMID- 3053344 TI - [Urodynamic studies in gynecology]. PMID- 3053345 TI - [Basic principles of human reproduction and its disorders]. PMID- 3053346 TI - [Endocrinology of normal and disordered early pregnancy]. PMID- 3053347 TI - [Ultrasound diagnosis in disordered early pregnancy]. PMID- 3053348 TI - [Infection as a complication in early pregnancy]. PMID- 3053350 TI - [Congenital and acquired organ changes of the uterus and habitual abortion]. PMID- 3053349 TI - [Clinical aspects of infection in early pregnancy]. PMID- 3053351 TI - [Environmental pollution as disorders of early pregnancy and as a cause of abortion]. PMID- 3053352 TI - [Immunodiagnosis and therapy of habitual abortion]. PMID- 3053353 TI - [Pathologico-anatomic findings in disordered early pregnancy]. PMID- 3053354 TI - [Psychosomatic aspects of abortion]. PMID- 3053355 TI - [Assessment and therapy in the clinic in repeated abortion]. PMID- 3053356 TI - Differences between beige and bg/+ mice in the disruption of plasma proteinase regulation in the tumor-bearing state or following Corynebacterium parvum treatment. Evidence for the involvement of polymorphonuclear leukocyte proteinases. AB - Mice bearing the B16 melanoma or treated with Corynebacterium parvum develop elevated levels of plasma neutral proteinase activity. Similar experiments carried out with C57BL/6-bg/bg (beige) mice, which are genetically deficient in polymorphonuclear neutrophil (PMN) proteinases, revealed that such mice develop significantly diminished elevation in plasma proteinase activity compared to C57BL/6-bg/+ mice. Lysates of C. parvum elicited PMN from beige mice contained approximately 80% less neutral proteinase activity as did lysates of PMN from bg/+ mice. These results indicate that host cells, such as PMN, may become activated during the tumor progression, or following C. parvum treatment, causing degranulation and a subsequent elevation in plasma proteinase levels. If such an interpretation is correct, then this phenomenon may be the murine corollary to what has been observed in patients with certain inflammatory diseases or tumors. PMID- 3053357 TI - Morphometry of human blood leukocyte ultrastructure: its potential value in haematology. AB - A review of the literature on ultrastructural morphometry of human blood leukocytes has been carried out. It is concluded that: (1) Blood leukocytes are particularly suitable for morphometric study. (2) Morphometric methods have proved valuable in defining differing cytological features of cells in various lymphoid malignancies, and in demonstrating ultrastructural differences (which could not otherwise have been detected) in monocytes and eosinophils corresponding to known functional changes. (3) Appropriate and valid numerical procedures are essential for determining morphometric equations and statistical probabilities; both morphometric measurement and statistical analysis are made easier by the use of computers. (4) Ultrastructural morphometry should ultimately find an important place in clinical haematology. PMID- 3053358 TI - Differentiation induction therapy of acute myelogenous leukaemias. AB - This article reviews the progress achieved in the field of inducing differentiation in human myeloid leukaemia in vitro and the need for applying this model in leukaemia therapy in vivo. The pre-clinical evaluation of differentiating agents in human myeloid leukaemia is analysed. The clinical experience with differentiation induction therapy in acute myeloid leukaemia is reviewed. This review examines the prospective application of this new form of therapy in patients with acute myeloid leukaemia. PMID- 3053359 TI - Role of actin polymerization in monocyte phagocytosis of yeast cells effect of cytochalasin B. AB - The role of the cortical actomyosin-like contractile system in monocyte phagocytosis was studied by means of inhibition with cytochalasin B. Monocyte phagocytosis of yeast cells was assayed with a fluorescence extinction technique of surface-located phagocytosis which distinguishes between adherence and ingestion phases of total phagocytosis. Cytochalasin B, 10 micrograms/ml, inhibited monocyte phagocytosis by 20-30%. The cytochalasin inhibition was significant and restricted to the ingestion phase. The finding supported the hypothesis that the contractile structures of the cell have a contributory role in IgG-mediated monocyte phagocytosis on a surface. It is suggested that the cytochalasin effect reflected the role of chemotaxis in monocyte phagocytosis assayed by the present technique. PMID- 3053360 TI - The interaction between fibrinogen and 3H-L-arginine cationic peptides derived from fibrosarcoma in the presence of thrombin. AB - It has been found that cationic protein breakdown product--3H-L-arginine labelled peptide fraction--interacts with fibrinogen in the presence of thrombin. The formation of the fibrin clot under these conditions makes the clot resistant to the fibrinolytic action of plasmin. PMID- 3053361 TI - [Inhalation injury in burn patients]. AB - The clinical course of inhalation injury is variable. The routine use of fiberoptic bronchoscopy provides an accurate and safe diagnosis; by this examination it is possible to recognize alterations down to the terminal airway. Biopsies of tracheobronchial mucosa can ensure the diagnosis. By means of fiberoptic examination and blood gases it is possible to realize a differentiation of inhalation injury. According to the analysis of our patients an inhalation injury was found in 20.1%. The mortality rate due to the inhalation injury amounts to 14.4%. PMID- 3053362 TI - [Multiple symmetrical lipomatosis. A retrospective study of 14 cases and review of the literature]. AB - "Multiple symmetric lipomatosis" is an accumulation of fatty tissue in upper areas of the body mainly effecting middle aged men. It is a rare benign disease connected with differing disorders. We report 14 male patients suffering from "multiple symmetric lipomatosis" and focus on the etiology and on associated disorders. Differential diagnosis, pathohistological considerations, clinical behaviour, and operative treatment are discussed. A review of the literature is presented. PMID- 3053363 TI - [Blood supply of free skin flaps]. AB - This article discusses the factors which determine the blood supply of free skin flaps. The method of free transfer consists of three steps, namely, the elevation of the flap, its transfer to the target area with microvascular anastomosis, and the healing of the flap on the recipient site. During the first two steps the flaps meet with a diminished blood supply and, in addition, their tissues undergo a period of ischemia. The flaps are usually able - within certain limits - to tolerate these disturbances of blood circulation. The healing of the flap begins with the proliferation of capillaries in the recipient site, which is stimulated by the relative hypoxia of the transferred tissues. The first capillary links between the recipient site and the flap can usually be observed on the third day after the transfer. On the sixth day after the operation, the anastomoses are quite numerous and can take over the blood supply to the flap. The progress of this dynamic process depends on the functional ability of the anastomosed vascular pedicle. The development of a new blood supply to the flap leads to profound remodelling of its original vascular system and the axial character of the flap is usually lost. PMID- 3053364 TI - [Laser Doppler flowmetry in vital bone transplantation. Experimental and clinical results]. AB - Laser Doppler Flowmetry allows the measurement of the blood flow rate of different types of tissue. Rib on a muscle pedicle was compared with rib on a vascular pedicle in an animal experiment. The morphological results were compared with the LDF-results. During eleven clinical transfers of iliac crest bone grafts and one osteocutaneous scapular flap the blood flow values were measured intraoperatively before and after the transplantation. Postoperative monitoring was possible for 48 hours with the endoscopic probe and a screw implant. The blood flow of vascularized bone transplants is excellent and even rises post operatively in some cases. PMID- 3053365 TI - Plasmid pIMI38--the pBR322 derivative with increased stability in E. coli cells. AB - Plasmid pIM138 which had been characterized by the higher resistance of its DNA replication to the action of clorobiocin in comparison with the progenitor plasmid, was tested for its stability in host cells in the absence of the antibiotic. Growing without selective pressure, pIM138 was better maintained in cells than pBR322. The stability in the presence and in the absence of clorobiocin can be unanimously assigned to the plasmid itself, but some influence of host cells cannot be excluded. PMID- 3053366 TI - Increased sterol formation in Saccharomyces cerevisiae. Analysis of cell components and ultrastructure of vacuoles. AB - In Saccharomyces cerevisiae nitrogen limitation under aerobic conditions (low specific growth rate) provokes an enhanced synthesis of sterols. Analysis of east cultures during the enhanced sterol biosynthesis showed a temporary decrease of protein content and a simultaneous increase in polysaccharide and lipid levels. This was reflected in the ultrastructure of cells where numerous lipid globules (spherosomes, oleosomes) appeared around extensive membrane-bound compartments containing membrane vesicles and lipoprotein material. Electronograms showed that such compartments were formed between the layers of endoplasmic reticulum and belonged to the vacuome phase of the yeast cell. It appears that vacuoles formed in yeast during enhanced synthesis of sterols have a storage rather than a lysosomal function. PMID- 3053367 TI - The genotoxic activity of glycerol in an in vitro test battery. AB - Glycerol, a widely distributed constituent of food and an additive used in cigarette manufacture, has been tested for genotoxic potential in a battery of short-term genotoxicity assays. Glycerol was evaluated in the Ames Salmonella typhimurium mutagenesis assay (strains TA98, TA100, TA1535, TA1537 and TA1538), in the rat hepatocyte unscheduled DNA synthesis assay, in the Chinese hamster ovary (CHO) chromosome aberration assay, the CHO sister chromatid exchange assay and the CHO mammalian mutagenesis assay. All assays (except the rat hepatocyte unscheduled DNA synthesis assay) were conducted both with and without the addition of Aroclor-induced rat liver S-9. The results of all tests were negative, showing that neither glycerol nor its metabolites have genotoxic activity in the battery of tests used. PMID- 3053368 TI - An hypothesis of the mechanism of urinary bladder tumorigenesis in rat ingesting sodium saccharin. AB - An hypothesis is presented of a mechanism for the sodium saccharin (NaS) associated tumorigenesis of the urinary bladder that occurs in male rats. The ingestion of high doses of NaS is associated with increased urine volume and bladder mass. In rats with an inherently high urine output, the diuresis associated with NaS ingestion combined with the increasing diuresis that occurs with age in male rats results in a chronic demand for a bladder-volume increase that is met by excessive cell division of the bladder epithelium. This enhanced mitosis in the bladder epithelium can result in a significant incidence of bladder tumours. Male rats exposed to NaS during early life show an exacerbation of tumour incidence, and it is proposed that this is because the exacerbation of the effects of NaS on the gastro-intestinal and urinary tracts results in increased urine output and bladder hyperplasia in these rats. PMID- 3053369 TI - Methylxanthines: toxicity to humans. 2. Caffeine. AB - While there are several comprehensive reviews on the toxic effects of methylxanthines in animals, data on the toxicity of these chemicals in humans has not been extensively reviewed in one document. In a previous paper (Stavric, Fd Chem. Toxic. 1988, 26, 541), the toxicity of theophylline was reviewed. This paper, the second of three, is intended to provide an overview of the human toxicity of caffeine. Only pertinent and recent information on caffeine toxicity is summarized. In addition, some information regarding the benefits of caffeine and the mechanism of its effects is also provided. The use, effects and toxicity of caffeine intake are reviewed separately for different segments of the population. Controversy concerning the possible association of caffeine with fibrocystic disease of the breast and over the behavioural effects of the drug is presented briefly. PMID- 3053370 TI - [Reactive capacity of animal and human condylar cartilage at the cellular and molecular levels in the light of a cybernetic concept of facial growth]. PMID- 3053371 TI - [Responsibilities of psychosomatic medicine in reproduction medicine. A report of experiences]. PMID- 3053373 TI - [Premenstrual syndrome should not be underestimated. Many women are affected- neither resignation nor polypragmatism are appropriate]. PMID- 3053372 TI - [Preventive use of antibiotics in colorectal interventions. Short-term versus one shot prevention--a multicenter study]. PMID- 3053374 TI - [Immunologic-cytologic monitoring following heart transplantation]. PMID- 3053376 TI - [Theophylline therapy: patient compliance and dosage. A clinical study]. PMID- 3053375 TI - [Sleep apnea--diagnosis, clinical aspects and therapy]. PMID- 3053377 TI - [Control of venous stasis. Use of low molecular weight heparin--a multicenter study]. PMID- 3053378 TI - [Calcium antagonist effective in migraine attack. Problems with ergotamine- sublingual administration is necessary for calcium antagonists]. PMID- 3053379 TI - [Radius shortening in lunate bone osteomalacia]]. PMID- 3053380 TI - The relationship between clinical and echocardiographic findings in mitral valve prolapse. AB - Incorporating prognostically related auscultatory, M-mode, 2DE and recent Doppler echocardiographic features, the following strict criteria for establishing the diagnosis of mitral valve prolapse (MVP) have been advanced: 1. auscultatory; mid to-late systolic clicks and a late systolic murmur at the apex or mid-to-late systolic clicks at the apex which move appropriately with maneuvers that alter LV volume or late systolic murmur at the apex in young patients (coinciding that a similar murmur in elderly population is non-specific for MVP); 2. two dimensionally "targeted" M-mode criterion: marked (greater than 3 mm) late systolic buckling posterior to C-D line (moderate 2 mm late systolic buckling or 3 mm holosystolic displacement "arouse suspicion" but do not establish MVP); 3. two-dimensional echocardiographic criteria: severe bowing of leaflet(s) on the parasternal long axis and four-chamber view (mild to moderate bowing alone are unacceptable) or left atrial coaptation point; 4. Doppler echocardiographic criteria: moderate or severe Doppler mitral regurgitation with any degree of leaflet bowing or mild Doppler mitral regurgitation with at least moderate bowing of one leaflet (mild leaflet bowing and mild mitral regurgitation can be regarded as "probable MVP"). The concept of mitral valve prolapse syndrome encompasses that which was earlier described in patients with a high prevalence of symptoms. In controlled studies, however, it has become apparent that cardiac and psychiatric symptoms can be found as frequently in normal subjects as in those with MVP. These results indicate that clinicians may have erroneously diagnosed patients with MVP because of premature acceptance that MVP is the cause of a distinctive syndrome.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3053381 TI - The problem of echocardiographic detection of mitral valve prolapse and determination of its true prevalence. AB - The prevalence of mitral valve prolapse and the frequency of associated complications is currently still not known with certainty. In post-mortem studies, myxomatous changes of the mitral valve are found in less than 5%. The relationship between characteristic auscultatory findings, detectable in 6 to 18% of young asymptomatic subjects, and angiographic criteria for mitral valve prolapse, observed in up to 30% and more of those undergoing routine cardiac catheterization, is similarly unclear. Establishing the diagnosis based on M-mode echocardiographic criteria has yielded problems, in particular, a frequency too high for apparently healthy subjects. By means of two-dimensional echocardiography, displacement of a mitral leaflet could be detected more frequently in the four-chamber view than in the parasternal long-axis view, a finding which renders both the diagnostic comparability and the assumption of a planar mitral annulus questionable. Accordingly, a saddle-shaped mitral annulus has been postulated. The hypothesis of the saddle-shaped form has been repeatedly tested and confirmed: on a valve model, in patients without mitral valve disease by means of two-dimensional echocardiography as well as by means of three dimensional reconstruction of two-dimensional echocardiographic images and, lastly, in animal experiments with surgical implantation of radioopaque markers with fluoroscopic observation. Patients can be divided into one of three categories according to the position of the leaflet with respect to the highest or lowest point of the mitral annulus: level of coaptation of the leaflets beneath, completely within or above the highest and lowest points of the annulus. A subgroup of patients in the latter category can be regarded as abnormal.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3053382 TI - Correlation between left ventriculography, auscultation, and M-mode and two dimensional echocardiography in mitral valve prolapse. AB - Mitral valve prolapse (MVP) is a common valvular abnormality which is observed in as many as 5% of the general population. Although invasive as well as noninvasive tools have been developed to determine the existence of this disorder, none is perfect and false negative as well as false positive diagnoses abound. Because MVP is a relatively benign disorder, it has also not been easy to make the usual clinical-pathological correlations. Left ventriculography is considered by many to be the gold standard, but this designation is probably not deserved. The angiographic criteria used by most do not permit unequivocal separation of normal mitral valve systolic bulging from pathologic MVP, and the interobserver and intraobserver variability of interpretation is high. However, false positive diagnoses can be eliminated if MVP is diagnosed only when para-annular displacement of mitral leaflet tissue is detected during systole rather than simple leaflet bulging. Although mid-systolic clicks and late systolic murmurs have proven to be the auscultatory hallmarks of this disorder, many patients have these signs without other diagnostic findings, consequently making it impossible to confirm the presence of MVP. Furthermore, the appearance of diagnostic echocardiographic abnormalities in patients with normal cardiac examinations implies that auscultation is not a sensitive marker of MVP. Both M-mode and two dimensional echocardiography have technical limitations and the repeatability of interpretation of these tests is disappointingly low (80 to 90%). Because of these difficulties the angiographic-echocardiographic correlation is only fair. Nonetheless echocardiography has generally been accepted as the diagnostic modality of choice. Future technical improvements will likely enhance the diagnostic accuracy of this technique. PMID- 3053383 TI - Progression of mitral regurgitation in patients with mitral valve prolapse. AB - Mitral valve prolapse (MVP) is a very common clinical entity which is frequently associated with mild mitral regurgitation (MR) and which most commonly becomes clinically manifest in the third and fourth decades of life. Severe MR associated with MVP, occurs much less frequently and is most commonly seen in patients above the age of 50 years. Relatively little information is available regarding the progression of mild to severe MR in patients with MVP. This report reviews a recent study which investigated the progression from mild to severe MR in patients with MVP. The study included 86 patients, average age 60 years, who presented with cardiac symptoms and severe MR. A high incidence of MVP was seen on echocardiograms (57 of 75 [75%]) and on left ventriculography (61 of 84 [73%]). Mitral valve replacement was performed in 75 patients. Pathologically all valves appeared grossly enlarged, severely floppy and had extensive myxomatous changes with collagen dissolution. 80 patients had a pre-existing heart murmur first detected at average age 34. Patients remained asymptomatic for an average of 25 years at which time clinical symptoms first appeared. After symptoms developed mitral valve surgery was necessary in most patients within one year. This rapid deterioration could partially be attributed to ruptured chordae in 39 of 76 patients (51%) or atrial fibrillation in 48 of 86 patients (56%). 28 patients had one or more serial clinical evaluations including auscultation, chest x-ray, echocardiography, and cardiac catheterization.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3053384 TI - [Mitral valve prolapse syndrome and arrhythmias]. AB - In patients with mitral valve prolapse syndrome various disturbances of cardiac rhythm can be observed such as atrial arrhythmias, ventricular tachycardias and conduction disturbances. Of timely interest are the questions of which etiology is at the basis of the arrhythmias, what is their relevance with respect to sudden cardiac death, what are the indications for treatment and which therapeutic results can be anticipated. Cardiac arrhythmias represent the most frequent complication of mitral valve prolapse. Holter ECG monitoring has assumed the central role in detection of all types of arrhythmias. As compared with normal persons, in patients with mitral valve prolapse, both ventricular and supraventricular arrhythmias can be found more frequently. Atrial arrhythmias: Supraventricular arrhythmias can be found less frequently than ventricular arrhythmias (Table 1). Premature atrial contractions can be observed in 35% of those with mitral valve prolapse but also in a similar number of normal individuals such that their presence is not of clinical relevance. There is only a tendency to more frequently incurred supraventricular couplets in mitral valve prolapse. Supraventricular tachycardias can be observed in 10.5 to 32% of which sinus tachycardia (heart rate greater than 120 beats per minute), paroxysmal atrial tachycardia and intermittent atrial fibrillation at about 5 to 6% each are not more common than in control subjects. Atrial fibrillation was seen more frequently in mitral valve prolapse with mitral regurgitation or, conversely, in mitral regurgitation due to mitral valve prolapse more frequently than in mitral regurgitation due to other causes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3053385 TI - Follow-up observations in patients with mitral valve prolapse. AB - The discrepancies reported in various studies with respect to long-term prognosis in mitral valve prolapse can be attributed to the criteria employed for diagnosis and to the differing patient populations studied. Furthermore, mitral valve prolapse is not a single, well-defined disease but reflects rather a broad spectrum of a disease process. Echocardiographically, patients can be identified with redundant, thickened leaflets with excessive mitral valve motion and prolapse as well as those with normal appearing leaflets but systolic prolapse representative of differing disease processes with differing prognosis. The incidence of sudden cardiac death is estimated at a low value of 1.9/10,000 patients per year. In the presence of a normal resting ECG, with no hemodynamically-meaningful mitral regurgitation and no evidence of redundant mitral leaflets the risk is even less. Cerebral embolic events occur with an estimated incidence of 1/6000 patients per year, similarly low; it can be assumed that patients with very myxomatous, redundant mitral leaflets have the highest risk. The incidence of infective endocarditis is also low, estimated at 1/5725 patients per year (0.175%). Risk factors for complications include: a systolic murmur, advanced age, male sex and leaflet redundancy. The most important complication is mitral regurgitation for which the incidence is highest in older men and in the presence of left ventricular, dilatation. PMID- 3053386 TI - HBV DNA and other hepatitis B virus markers in sera from long-term hemodialysis and kidney transplant patients. AB - Sera from 191 long-term hemodialysis patients and from 115 renal transplant patients were studied for the presence of HBsAg and other HBV markers. In 19.1% of the hemodialysis patients and 28.7% of the transplant patients, the sera were positive for HBeAg.HBV DNA in sera was detected by molecular hybridization. HBV DNA was present in the sera of 15 out of 16 hemodialysis patients positive for HBeAg, and in one hemodialysis patient positive for anti-HBe. All renal transplant patients positive for HBeAg were also positive for HBV DNA. Twelve transplant patients were positive for HBV DNA, but negative for both HBeAg and anti-HBe. Determination of HBV DNA was the most sensitive marker of infectivity in patients with end-stage renal disease, and in renal transplant patients. PMID- 3053388 TI - Whole body hyperthermia of rats decreases insulin binding to erythrocytes. AB - 125I-insulin binding to rat erythrocytes was studied to investigate the effect of whole body hyperthermia on the insulin receptor. Heat treatment of rats at 42 degrees C for 15 min caused a significant decrease (48.7% of control) in 125I insulin binding to rat erythrocytes. Scatchard analysis showed that the decreased binding resulted from a decrease in the number of the insulin receptors rather than from a decrease in receptor affinity. The decreased receptor number for insulin showed no evidence of recovery, 2 h and 8 h after the hyperthermia. Plasma insulin levels remained lower than the control, up to 8 h after the hyperthermia, whereas plasma glucose, which decreased immediately after the hyperthermia, increased higher than the control, 8 h after the hyperthermia. The low plasma insulin level and decreased number of insulin receptor are believed to be possible factors for the elevation of plasma glucose. PMID- 3053387 TI - Diabetic embryopathy and fuel-mediated organ teratogenesis: lessons from animal models. AB - The growing recognition that faulty maternal metabolism during early organogenesis may be implicated in the increased incidence of birth defects in pregnancies complicated by diabetes has prompted worldwide efforts to institute improved preconceptional metabolic regulation. However, the failure to identify the periods of greatest risk for diabetic embryopathy, the mediating teratogen(s), and the underlying mechanisms have complicated attempts to establish precise therapeutic guidelines and targets. Some of the reported in vivo and in vitro experiences with rodent models have been reviewed to derive relevant insights. Substantial literature indicates that diabetes (experimental as well as spontaneous) in pregnant rats and mice is attended by retardation of growth and developmental delay during embryogenesis, and a variable incidence of birth defects. Poor metabolic regulation of the diabetic mother during early organogenesis may also be followed by subsequent resorption of the conceptus at the site of implantation. Vulnerability to diabetes-related resorptions and all other forms of embryopathy appears to begin during the early postimplantation period and is greatest near the onset of neurolation. Overall susceptibility is markedly influenced by genetic factors and may be modified by the antecedent metabolic exposures of the conceptus ("carry-over effects"). Mediation for the anomalous embryo development in pregnancies of diabetic rodents appears to be multifactorial; all the aberrant fuels and fuel-related components of "the diabetic state" (e.g. high glucose; ketones; somatomedin inhibitor(s); osmolality, etc.) which have been tested to date display dysmorphogenic potential ("fuel-mediated organ terato-genesis") in vitro. All tissues in the conceptus appear to be at risk. Dose-response relationships for the individual metabolic teratogens may be influenced by additive and synergistic interactions so that the integrated possibilities cannot be assessed fully by measurements confined to a single fuel or fuel-related component. In the context of the day-to-day variability in diabetes "control" of the poorly regulated mother, and the relatively longer duration of organogenesis, these multifactorial possibilities may account for the multiple birth defects that can occur in individual offspring, and the seemingly non-specific pattern of diabetic embryopathy. Insulin therapy diminishes the dysmorphogenic effects of "the diabetic state" in rodents with experimental or spontaneous diabetes.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3053389 TI - Interrelation between glucose, insulin, C-peptide and 3-hydroxybutyrate in plasma and amniotic fluid in last trimester diabetic women without residual betacell function. AB - The relationship between maternal plasma and amniotic fluid (AF) concentrations of glucose, insulin, C-peptide and 3-hydroxybutyrate (3-HB) was analysed between 45 to 140 minutes after a standardized breakfast in 8 type I diabetic women without residual betacell function and in 13 nondiabetic control women during the last trimester of gestation. AF levels of both glucose and C-peptide were slightly and AF insulin levels significantly (P less than 0.05) elevated above normal in the diabetic women. 3-HB levels in plasma and in AF were significantly (P less than 0.05) elevated in the diabetic group between 45 to 65 minutes after breakfast. AF insulin and glucose was significantly correlated in the diabetic group (r = 0.96, P less than 0.05). During the 2 hour study period AF levels of glucose, insulin and C-peptide remained essentially unchanged in both groups of women. Changes in maternal plasma 3-HB concentrations seemed to be more rapidly reflected in AF. PMID- 3053391 TI - Tough strategies boost multis' profits. PMID- 3053390 TI - Metoclopramide-induced changes in plasma aldosterone "in vivo" are independent from renin-angiotensin system activity. PMID- 3053392 TI - HCFA: teaching hospitals should seek donations. PMID- 3053393 TI - Hsiao's relative value scale shifts MD incomes, but not utilization. PMID- 3053394 TI - HCFA regulations will identify substandard care. PMID- 3053395 TI - New payment system squeezes NYC hospitals despite high occupancy. PMID- 3053396 TI - Ultrasound opens windows. PMID- 3053397 TI - HCFA eager to crack down on patient dumping. PMID- 3053399 TI - Drug industry responds to new buying strategies. PMID- 3053398 TI - Hospitals big losers with new cataract rate. PMID- 3053400 TI - Changes in home care benefits are imminent. PMID- 3053401 TI - Medical practice guidelines may affect payment. PMID- 3053403 TI - Larger issues at stake in Medicaid suits. PMID- 3053402 TI - Medicare balances rise, but days in receivables drop. PMID- 3053404 TI - States, hospitals aim to cut costly ED visits. PMID- 3053406 TI - Coronary artery surgery--a changing profile. PMID- 3053405 TI - Diagnostic reliability of endomyocardial biopsy for assessment of cardiac allograft rejection. AB - Endomyocardial biopsy (EMB) of the right ventricle has demonstrated clinical use primarily for the detection of allograft rejection. Since its introduction, the procedure has been demonstrated to be safe and adaptable to an ambulatory population. The detection and grading of rejection have relied heavily on the criteria originally proposed by the Stanford group. Similar criteria are under study for diagnosis of myocarditis. Using autopsy-acquired cardiac allograft specimens, we report the reliability of EMB for grading of rejection and establish the dependence on the morphologic features used as the diagnostic cut off and the fragment number obtained at biopsy. The reliability of an interpretation protocol based on the pattern of inflammation present in the multiple fragments submitted is established. We compare the reliability for rejection prediction between this approach and the currently accepted approach based on the presence of myocyte necrosis. PMID- 3053407 TI - Consideration of a functional role of an extracellular sialidase secreted by cultured fibroblasts. PMID- 3053408 TI - Recent advances in ganglioside metabolism. PMID- 3053409 TI - The human red cell sialoglycoprotein, glycophorin A: biosynthesis, glycosylation and interaction with external ligands. PMID- 3053410 TI - Endogenous factors determining the agglutinability of erythrocytes with concanavalin A. PMID- 3053411 TI - Recent advances in the chemistry and technology of gangliosides. PMID- 3053412 TI - Red cell membrane alterations in malaria. PMID- 3053413 TI - Studies on a lectin from Saccharomyces cerevisiae. PMID- 3053414 TI - Malignancy-related cell surface mucin-type glycoproteins. PMID- 3053415 TI - Glycolipids, glycoproteins and membrane fusion. PMID- 3053416 TI - Receptor binding domain of glycophorin A for Plasmodium falciparum surface proteins. PMID- 3053417 TI - Tumours de parotid gland--a review of 88 patients & current methods of treatment. PMID- 3053418 TI - Carcinoma of breast and acute myeloid leukemia in the same patient: report of a case and review of literature. PMID- 3053419 TI - Signet ring cell lymphoma simulating liposarcoma--a case report with brief review of literature. PMID- 3053420 TI - [Vasculitis and granulomatosis in the respiratory tract]. AB - Lately new aspects of diseases have been described in long known forms of pulmonary vasculitis and granulomatosis like Wegener's granulomatosis (WG), sarcoidosis (SA), exogenous allergic alveolitis and the Churg-Strauss syndrome. In WG it is clinically important to distinguish between two different phases of disease - the early initial phase with localized inflammation and granuloma formation and the later generalized form of disease where an involvement of virtually every organ system can occur. Recently autoantibodies (antineutrophil cytoplasmic antibodies) have been discovered which are highly specific for WG and additionally are helpful to determine the activity of the disease. With respect to both findings and more recent experience with various therapeutic regimes a therapy considering the phase and activity of disease is recommended. In SA the analysis of the bronchoalveolar lavage has not only led to a immunodiagnostic concept but also given immunopathogenic insights. In contrast to former ideas the SA is not the consequence of an anergic state but more likely of an increased activity of the immune system which is hinted at by the finding of activated CD4 positive lymphocytes in the interstitial tissue and lavage fluid. In exogenous allergic alveolitis new disease concepts are arising from the better understanding of the triggering agents which can now be better analysed. PMID- 3053421 TI - [Histomorphology and immunopathology of granulomatous inflammation]. AB - Every inflammation has its morphological substrate, however, not every morphological substrate is specific for a particular disease. On one side, this is due to the limited reaction spectrum of the organism, on the other side, to the fact that completely different etiological agents are able to activate the same pathomechanism. Since this holds true for granuloma formation also, the term "specific inflammation" has to be restricted considerably today. On the other hand, recent progress in the field of mediators of inflammation frequently permits a pathogenetic classification of granulomatous inflammatory processes - even before the etiology has been clarified (e.g. Morbus Boeck). In addition, the possibility to identify "granuloma cells", in particular macrophage- and lymphocyte-subpopulations, in situ by immunohistological means (by monoclonal antibodies to surface markers) expands the scope of conventional histomorphological techniques significantly. PMID- 3053422 TI - [Particle agglutination test: a new technic for the detection of HIV-1 antibodies]. AB - Two techniques for detection of anti-HIV-1 antibodies were comparatively investigated. It is shown that the particle agglutination (PA) is at least equivalent to the enzyme immunoassay (EIA) as far as sensitivity is concerned. A ten-fold increased sensitivity of PA was found when compared with Western Blot results. Furthermore, the detection of specific IgM antibodies in sera of HIV-1 infected persons is possible when the PA is used. The high stability of PA reagents when stored at 37 degrees C, the simple performance of the assay, and the possibility of automatization are important advantages of the PA when compared with the EIA. PMID- 3053423 TI - Suppressive role of NK cells in pokeweed mitogen-induced immunoglobulin synthesis: effect of depletion/enrichment of Leu 11b+ cells. AB - Natural killer (NK) cells probably have immunoregulatory effects. However, the evidence to date is mainly based on the suppressive effect of enrichment with relatively impure NK populations (large granular lymphocytes, LGL, Leu 7a+ cells). Here we report on the effect of enrichment and depletion of Leu 7a+ and Leu 11b+ cells (the latter containing virtually all NK activity in freshly prepared lymphocytes) on pokeweed mitogen (PWM)-induced immunoglobulin (Ig) synthesis. Enrichment suppressed Ig synthesis to a degree dependent on the number of cells added, and was not enhanced further by their pretreatment with interferon. Furthermore, depletion of Leu 11b+ cells from peripheral blood lymphocytes (PBL) led to marked enhancement (2-25-fold increase) of Ig synthesis, suggesting these cells may normally exert a suppressive effect. The possible underlying mechanisms were investigated further. Enhanced Ig synthesis by Leu 11b depleted cultures was associated with an increased number of Ig-secreting cells by plaque assay, but with no change in numbers of CD4+ or CD8+ cells. Treatment of PBL with monoclonal antibodies (anti-Leu 7a/Leu 11b) alone suppressed PWM induced immunoglobulin synthesis. We conclude that NK cells play a role in the regulation of Ig production, at least in part by an effect on activation/differentiation of B cells, but independent of altered T-cell subpopulations. The effect may be unrelated to their cytotoxic function (being unaffected by interferon, IFN), although the direct effects of anti-Leu 11b and Leu 7a in enhancing the suppressive effect suggest an alternative activation pathway. PMID- 3053424 TI - Adoptively transferred experimental allergic encephalomyelitis in chimeric rats: identification of transferred cells in the lesions of the central nervous system. AB - Experimental allergic encephalomyelitis (EAE) was induced by adoptive transfer of myelin basic protein (MBP)-activated LEW spleen cells into (LEW x PVG/c) F1--- LEW chimeras. By double-immunofluorescent staining using OX27, which is specific for RT1c, and monoclonal antibodies (mAb) against various T-cell antigens (TAg), inflammatory cells in the lesions of the central nervous system (CNS) were categorized into MBP-activated and transferred LEW T cells (TAg+ OX27-), accompanying T cells (TAg+ OX27+) of chimera origin and non-T cells (TAg- OX27+). Examination of the lesions at various stages of EAE revealed that transferred OX19 (CD5)+ T cells accounted for 46% of the total number of inflammatory cells at the preclinical stage, became reduced to 23% at the clinical stage and recovered to a level between those of the preclinical and clinical stages at the recovery stage. In parenchymal infiltrates, 93% of the total T cells were transferred cells at the preclinical stage, whereas 66% were present in perivascular aggregates. At the clinical stage, the proportion of transferred T cells in the parenchyma was not different from that in the perivascular cuffs. At the recovery stage, the proportion of transferred T cells in the parenchyma was increased. Collectively, MBP-activated and transferred T cells first appeared in the CNS parenchyma followed by infiltration of T and non-T cells of recipient (chimera) origin. All these inflammatory cells formed the lesions of full-blown EAE. At the recovery stage, inflammatory cells decreased in number in all the compartments of the CNS. Transferred T cells formed the major proportion of parenchymal infiltrates at this stage. These findings strongly suggest that transferred T cells remain in the CNS parenchyma longer than cells of chimera origin and that antigen-activated T cells have well-expressed CNS-homing activity. PMID- 3053425 TI - Stimulation of liposome-induced humoral immune responses by non-ionic block polymer surfactants in Xid mice. AB - Non-ionic block polymers (NBPs) have proved to be potent adjuvants for the humoral immune response against liposomes haptenated with tripeptide-enlarged dinitrophenyl groups (hapten J). Since both reversed triblocks and normal octablocks displayed adjuvant activity, reversed octablocks, in which structural properties of both groups are combined, were also tested for their adjuvant activity. The latter compounds displayed very strong adjuvant activity for J haptenated liposomes, not only in normal BALB/c but also in (CBA/N x BALB/c)F1 progeny. To test the applicability of NBPs as adjuvants in semi-synthetic vaccines, the capacity of NBPs to stimulate the immune response against liposomes haptenated with Streptococcus pneumoniae type 3 capsular polysaccharide-derived oligosaccharides was analysed. In these studies, again NBPs proved potent adjuvants, stimulating antibody production to a large extent. In male (CBA/N x BALB/c)F1 mice, which carry a X-chromosome-linked immunodeficiency (Xid), antibody levels were stimulated to the largest extent by a normal octablock. Stimulation of antibody titres, however, did not result in increased protection in these Xid mice. PMID- 3053426 TI - Antigenicity of the carboxyl terminus of insulin: isolation of human insulin specific monoclonal antibodies. AB - Monoclonal technology was used to isolate antibodies binding the B30 (carboxy) terminal residue in the polyclonal-provoked immune response to human insulin. Although both spleen and lymph node cell fusions were carried out, only the latter were successful in isolating monoclonal antibodies that bound the carboxy terminal of human insulin. The binding of such antibodies was abolished or diminished by substitutions of the B30 residue. Studies with insulin species variants showed that the molecular binding between antibody and insulin may be critically determined by a subresidue feature, e.g. presence or absence of a single methyl group, as shown by the binding of the monoclonal antibody D10 to human insulin (threonine at B30) but not to rabbit insulin (serine at B30). Such studies are of interest in the study of the molecular basis of antibody-antigen interaction. PMID- 3053428 TI - Somatic mutations in B lymphocytes: new perspectives in tolerance research? AB - This paper extends the concept of clonal anergy developed in the author's laboratory. It has been shown that the primary population of B lymphocytes induced into clonal expansion and IgM antibody formation by mitogens contains many cells capable of autoantibody synthesis, but the affinity of binding to the self constituents, or indeed to foreign antigens, is low. The creation of high affinity antibody, which will still register strongly in an ELISA as an IgG molecule, demands not only the addition of lymphokines to cause isotype switching, but also intentional immunization of the donor mice to permit mutations in V region genes and selection of higher affinity B memory cells. This process appears to begin about 6 days after in vivo immunization. It is postulated that these mutational events occur primarily in germinal centres, and that there must be mechanisms to prevent escape of cells which, by chance, mutate not to higher affinity against an immunogen, but to higher affinity against a self constituent. If such mutants were allowed to enter the long-lived, recirculating pool of B lymphocytes, they might pose a graver threat of autoimmune disease than the low-affinity anti-self cells of the primary repertoire. Therefore, it is suggested that recently mutated germinal centre B cells represent a pool of 'pre-memory' cells, which are immature in the sense of displaying the same kind of sensitivity to negative signalling by antigen that immature B cells from newborn spleen or adult bone marrow display. If so, then the earliest phases of memory generation represent a second window of opportunity for tolerance induction within the B lymphocyte compartment. PMID- 3053429 TI - Festschrift: To honour Professor Derrick Rowley. PMID- 3053427 TI - Characterization of a 95,000 molecule on sheep leucocytes homologous to murine Pgp-1 and human CD44. AB - The phagocyte glycoprotein-1 (Pgp-1) antigen of mice is a 94,000 MW molecule with a wide tissue distribution, but no attributed function. We produced a monoclonal antibody (mAb) termed 25-32 which recognizes the Pgp-1 molecule of numerous mammalian species, including humans and sheep. Preclearing experiments with I42/5, a rat anti-mouse Pgp-1 mAb that cross-reacts with human Pgp-1, established the specificity of 25-32 for human and sheep Pgp-1. Moreover, an antibody recognizing human CD44, termed F10-44-2, also reacted with the same molecule as that recognized by 25-32 and I42/5, so establishing the co-identity of CD44 and Pgp-1. Within the sheep thymus, Pgp-1 was expressed most strongly by medullary thymocytes and stromal cells, and by small numbers of cells at the subcapsular cortex. Pgp-1 was expressed early in thymic ontogeny; all 35-40-day gestation fetal sheep thymocytes were intensely Pgp-1+, but by 80 days the number of reactive thymocytes had decreased to adult levels. The expression of Pgp-1 on lymphocytes was markedly increased after stimulation with mitogens, or with phorbol esters and ionomycin. The highly conserved nature of Pgp-1 through evolution, its expression on virtually all cell types within the body, and its increased expression on rapidly dividing cells indicate that this molecule mediates an important function, possibly serving as a hormone or metabolite receptor. PMID- 3053430 TI - [Chronic non-hereditary dermo-epidermal bullous dermatoses in the child. Immunopathologic and case review]. PMID- 3053431 TI - [Onychopathy in the renal transplantation patient]. PMID- 3053432 TI - [Association of psoriasis and bullous pemphigoid. Clinical case]. PMID- 3053433 TI - [Cutaneous histopathological changes in uremic patients. Preliminary report]. PMID- 3053434 TI - Induction and rapid screening of monoclonal antibodies against cyclosporin A. AB - Cyclosporin C-hemisuccinate was coupled to thyroglobulin by the mixed anhydride method. Cyclosporin C was used instead of cyclosporin A (CsA) because of lack of functional groups of CsA. The resulting protein-cyclosporin C conjugate allowed us to induce high antibody titers also against cyclosporin A in rabbit and mice. Polyclonal and monoclonal antibodies were prepared following standard procedure. Since no standard methods for screening and quantification of anti-CsA-antibodies were available, two methods were adapted: (a) liquid phase radio assay (RA) and (b) solid phase enzyme-linked immunoassay (ELI-SA). For the former procedure inactivated charcoal was applied to separate the antibody-bound and the unbound CsA. CsA-coated PVC microtiter plates were used for the latter. PMID- 3053435 TI - Induction in human allograft recipients of unresponsiveness to anti-lymphocyte globulin. AB - The observations presented here confirm previous reports that polyclonal ALG prepared at the University of Minnesota or ATGAM of The Upjohn Co., administered as described, rarely induced sensitization of patients to the horse gamma globulin. In addition, the phenomena of transient antibody production prior to the onset of unresponsiveness and the induction of unresponsiveness in individuals with preexisting antibodies were observed. PMID- 3053437 TI - Antibody response to lipopolysaccharide antigen in Shigella dysenteriae type 1 infection. PMID- 3053436 TI - A comparative study of 50% alcohol and ethanolamine oleate for oesophageal sclerotherapy. PMID- 3053439 TI - Serological study of patients clinically suspected to have toxoplasmosis in Kashmir. PMID- 3053438 TI - Comparison of ELISA with metabolism inhibition test for detection of Ureaplasma urealyticum antibodies in nongonococcal urethritis. PMID- 3053440 TI - Xylenes: uses, occurrence and exposure. PMID- 3053441 TI - Past and present exposure determination techniques for benzene, toluene and xylene. PMID- 3053442 TI - Sampling and analysis of industrial air. PMID- 3053443 TI - Sampling and analysis of ambient and indoor air. PMID- 3053444 TI - Sampling and analysis of drinking-water and food. PMID- 3053445 TI - Genetic effects of benzene, toluene and xylene. PMID- 3053446 TI - Biological monitoring of exposure to benzene, toluene and xylene. PMID- 3053447 TI - Carcinogenicity of benzene, toluene and xylene: epidemiological and experimental evidence. AB - Benzene. The evidence for carcinogenicity of benzene in humans was evaluated by the IARC in 1982 as follows: "It is established that human exposure to commercial benzene or benzene-containing mixtures can cause damage to the haematopoietic system, including pancytopenia. The relationship between benzene exposure and the development of acute myelogenous leukaemia has been established in epidemiological studies. "Reports linking exposure to benzene with other malignancies were considered to be inadequate for evaluation. "There is sufficient evidence that benzene is carcinogenic to man." This evaluation now warrants some elaboration and updating. While the epidemiological evidence concerning benzene carcinogenicity is strongest for acute myelocytic leukaemia, there is some limited evidence of increased risks of chronic myeloid and chronic lymphocytic leukaemia. In addition, recent studies have suggested an increased risk of multiple myeloma, while others indicate a dose-related increase for total lymphatic and haematopoietic neoplasms. Corroborative evidence for such a generalized effect comes from experimental studies showing that exposure to benzene depresses all lympho-haematopoietic cell lines. While only limited evidence of benzene carcinogenicity in experimental animals exists, the recent findings of the National Toxicology Program (NTP, 1984) in the U.S.A. and Maltoni et al. (1985) strongly indicate that benzene is an experimental carcinogen. Toluene and xylene. While no direct human evidence is available, there is recent evidence of carcinogenicity of toluene and xylene at high concentrations in experimental animals. It should also be noted that any future epidemiological observations of cancer risks associated with toluene or xylene would have to take account of the suspected effects of benzene impurities. PMID- 3053449 TI - Benzene: uses, occurrence and exposure. PMID- 3053448 TI - Toxicokinetics of benzene, toluene and xylenes. PMID- 3053450 TI - Toluene: uses, occurrence and exposure. PMID- 3053451 TI - Tumor necrosis factor (cachectin) mediates induction of cachexia by cord factor from mycobacteria. AB - The mechanism by which cord factor (CF), a toxic glycolipid from mycobacteria, induces cachexia was studied in BALB/c mice. Body weight was markedly reduced 48 h after CF administration; the animals became severely wasted and exhibited hypertriglyceridemia, hypoglycemia, and high levels of tumor necrosis factor (TNF) in plasma. After CF administration, a transferable factor which caused cachexia and hypertriglyceridemia in recipient mice was detected in the blood. Dexamethasone partially inhibited the cachexia-inducing action of CF. Conditioned medium from adherent peritoneal cell cultures incubated with CF produced the same wasting symptoms when inoculated intravenously into mice. These studies also demonstrated that adherent peritoneal cells produced a humoral factor in response to CF which was related to CF-induced cachexia. Antiserum to recombinant TNF alpha prevented the cachectin action in passive-transfer experiments. Our findings indicate that cachectin (TNF) plays a role as a central mediator of the wasting induced by CF. PMID- 3053452 TI - Role of Shiga toxin in the pathogenesis of bacillary dysentery, studied by using a Tox- mutant of Shigella dysenteriae 1. AB - A Tox- mutant of Shigella dysenteriae 1, SC501, was genetically engineered by cloning the Shiga toxin operon, inserting a cassette into the A subunit gene, and exchanging this in vitro-mutagenized sequence with the wild-type gene. SC501 produced a low amount of residual cytotoxicity which was not neutralized by a rabbit immune serum directed against Shiga toxin. Invasion of cultured cells demonstrated that Shiga toxin had no effect on the rate of intracellular growth of bacteria or on the rapid killing of invaded host cells. On the other hand, several significant differences were observed in macaque monkeys infected intragastrically with either the wild-type strain or its mutant. The production of Shiga toxin by the invading strain was correlated with the presence of blood within stools, a sharp drop in blood polymorphonuclear cells, and histopathological alterations, such as the destruction of capillary vessels within the connective tissue of the colonic mucosa, severe inflammatory vasculitis of the peritoneal mesothelium, and major efflux of inflammatory cells to the intestinal lumen. It is proposed that Shiga toxin influences the severity of bacillary dysentery by inducing colonic vascular damage, which accounts for bloody stools, intestinal ischemia, and inflation of a polymorphonuclear intestinal compartment during the infectious process. PMID- 3053453 TI - Response of the murine hematopoietic system to chronic infection with Mycobacterium lepraemurium. AB - Mycobacterium lepraemurium infection of mice produces a chronic lethal disease that is characterized by massive accumulation of macrophages throughout the mononuclear-phagocyte system. We studied the influence of M. lepraemurium infection on the composition and function of the hematopoietic system. Medullary erythropoiesis was virtually abolished, as reflected by a small number of erythroid elements and a decrease in the number and frequency of erythroid progenitors in the bone marrow, together with reduced uptake of 59Fe into bone marrow hemin. On the other hand, erythropoiesis was observed in the spleen, as demonstrated by a large number of erythroid cells, a sixfold increase of 59Fe uptake, and a pronounced increase in the number of erythroid progenitors. A considerable increase of monocyte progenitors was observed in the spleen, and a more modest increase was observed in the bone marrow. This increase may be accounted for, at least in part, by greatly increased levels of macrophage-colony stimulating factor in the serum of infected mice. Thus, M. lepraemurium infection produces important changes in the hematopoietic system, during the course of which the spleen becomes the major hematopoietic organ. PMID- 3053454 TI - Killing of blood-stage Plasmodium falciparum by lipid peroxides from tumor necrosis serum. AB - The multiplication of Plasmodium falciparum in culture, as measured by [3H]hypoxanthine incorporation, was inhibited in a dose-dependent manner by rabbit tumor necrosis serum. The regimen by which tumor necrosis serum is produced caused significant increases in the levels of triglycerides and lipid peroxides, with the latter being indicated by the level of malondialdehyde in the serum. When tumor necrosis serum was depleted of lipoproteins by aerosil (fumed silica), no parasiticidal activity remained, and when it was separated by ultracentrifugation, more than 70% of the parasiticidal activity was found in the lipoprotein fraction. This suggests that lipid peroxides may account for most of the parasiticidal activity in tumor necrosis serum but that a nonlipid toxic factor may also be present. PMID- 3053455 TI - Expression of the histidine-rich protein PfHRP1 by knob-positive Plasmodium falciparum is not sufficient for cytoadherence of infected erythrocytes. AB - Six culture-adapted knob-positive Plasmodium falciparum parasites, four of which were nonbinding in an in vitro cytoadherence assay, were tested for the presence of the knob-associated histidine-rich protein PfHRP1. Two knob-positive, strongly cytoadherence-positive P. falciparum strains from Aotus monkeys were also examined. All parasites expressed PfHRP1, suggesting that it plays a structural or functional role related to knobs but is not by itself sufficient for cytoadherence of P. falciparum-infected erythrocytes. PMID- 3053456 TI - Expression of tumor necrosis factor in vitro by human mononuclear phagocytes stimulated with whole Mycobacterium bovis BCG and mycobacterial antigens. AB - Four mycobacterial preparations directly stimulated human blood monocytes and alveolar macrophages to produce tumor necrosis factor (TNF). Monoclonal antibody against TNF blocked (99%) TNF activity. Lipopolysaccharide was not responsible for the TNF activity generated; activity was unaffected in the presence of polymyxin B. PMID- 3053457 TI - The effects of infection prevention regimens on early infectious complications in marrow transplant patients: a four arm randomized study. AB - Three hundred and forty-two patients with hematological malignancies underwent allogeneic marrow transplantation from family donors and were allocated to receive 1) no specific infection prophylaxis in a conventional hospital room (control, 100 patients), 2) prophylactic systemic antibiotics (PSA) in a conventional hospital room (PSA group, 101 patients), 3) decontamination and isolation in a laminar air flow (LAF) room (LAF group, 65 patients) and 4) PSA in an LAF room (LAF+PSA group, 76 patients). Patients were studied for bacterial and fungal complications from the day of admission and until engraftment. LAF isolation was discontinued before engraftment in 27% (LAF+PSA group) to 32% (LAF group) of isolated in 26% (LAF+PSA group) to 27% (PSA group) of patients on prophylactic antibiotics. Septicemia occurred in 41%, 22%, 25% and 10% of patients in the control, PSA, LAF and LAF+PSA group, respectively. The incidence of septicemia was significantly less in the LAF+PSA group than in the control and LAF group with the incidence of septicemia significantly higher in the control group than in any of the other three groups. No other risk factors analyzed in proportional hazards regression tests were associated with septicemia acquisition. It is concluded that effective infection prevention modalities significantly reduce infection morbidity in transplant patients. Since most granulocytopenic transplant patients not receiving PSA will receive empiric or therapeutic broad spectrum antibiotics. The use of PSA in or out of LAF isolation is recommended as an effective modality to reduce septicemia acquisition. PMID- 3053458 TI - Corynebacterium group D2 and urolithiasis in a boy with megacalycosis. AB - This is report on a boy with megacalycosis in whom infectious urolithiasis after eradication of Proteus mirabilis was maintained by Corynebacterium group D2. PMID- 3053459 TI - Rapid diagnosis of infections caused by respiratory syncytial virus. PMID- 3053460 TI - Energy metabolism--an overview. AB - The most basic requirement for sustaining life is energy. Over the centuries it has been recognized that the living body is warm and begins to cool at the time of death. Referring to great scientists--Harvey, Boerhaave, Black, Priestley, Scheele, Lavoisier, Liebig, Pettenkofer, Rubner, Voit, Lusk, DuBois--the change in paradigm connected with the concepts of 'life', 'substrate intake' and 'body heat' and their underlying natural phenomena is outlined. The late 19th and early 20th century were extremely eventful in the evolution of the understanding of energy and its application to medicine. Definitions and concepts of metabolic body size, diet-induced thermogenesis, and physical activity are described. Further, metabolism adapted to starvation, postaggression metabolism and energy balance corresponding to their historical development and to the recent knowledge are elucidated. PMID- 3053461 TI - Awareness and assessment of periodontal problems among dentists and the public. AB - Awareness of periodontal problems by the public and their assessment by the practising dentist affect the levels of periodontal health. Many people do not recognize the symptoms of periodontal disease nor do they associate existing symptoms with the disease. Thus they do not perceive a need for professional or self-care. Most people in industrialized nations clean their teeth daily and visit the dentist at least once every 2 years. Yet, evidence suggests that levels of plaque and the incidence of bleeding gums are higher than expected either because people have not received individual instruction or they are not complying with appropriate regimens. Further, evidence suggests that many general dentists have low interest in the aetiology, prevention and treatment of periodontal diseases. It appears that such dentists have not retained their skills in the early recognition and diagnosis of periodontal diseases, nor in oral hygiene education or skill transfer. Likewise, only a small proportion of the general dentist's time is spent on prevention and periodontal care. To improve periodontal health, recommendations for the dental profession to pursue at the community, professional and individual levels are offered. PMID- 3053462 TI - Changing concepts of periodontal treatment: surgical and non-surgical. AB - Periodontal disease can be successfully treated by non-surgical procedures including supra- and subgingival scaling and root planing with hand or ultrasonic instruments. This non-surgical treatment is recommended for the initial management of all cases with the institution of a plaque control programme. Such management should be the basis of all periodontal therapy. The results obtained are mainly dependent on the skillfulness of the operator and the morphology of the pockets as well as upon patient related factors. Additional treatment, i.e. periodontal surgery, may be necessary if resolution of subgingival inflammation is not obtained. Pocket elimination by the use of surgical procedures (gingivectomy, flap operation with bone surgery) may be preferred in regions of the mouth where the aesthetic result is unimportant and where the removal of alveolar bone does not jeopardize the periodontal support of neighbouring teeth. In situations with deep infrabony pockets especially in anterior regions and in furcation areas a reconstructive surgical technique is the therapy of choice. Recent research in the field of guided tissue regeneration may, in the future, change the traditional approach to periodontal surgery. PMID- 3053464 TI - Review of methods of identification of high caries risk groups and individuals. Federation Dentaire Internationale Technical Report No. 31. AB - The search for methods of predicting dental caries activity began during the last century. The purpose of this review was to update the report on methods of caries prediction which resulted from the 1977 workshop, sponsored by the National Institutes of Health, Washington, DC, and also to identify the methods most likely to provide effective prediction of caries risk which should be given high priority in future research. The factors that need to be considered in assessing the value of a method of predicting caries risk are the correlation coefficient between the predictions and the final caries scores and in particular an assessment of the ability of the method to recognize subjects who will develop caries (sensitivity) and to exclude those who will not (specificity). The predictive power of the method should also be known. There is, however, the risk that when predictive tests are applied to a population with decreasing caries prevalence the number classified as false positive could be increased. This may limit the cost-effectiveness of preventive technique. The requirements of a good method of predicting dental caries are that the method should be simple, inexpensive and rapid and should identify subjects who will become diseased and exclude subjects who will remain healthy. To date, a wide variety of factors have been considered in the search for an effective method of predicting caries risk, but only a few have had some success. Certain epidemiological methods have shown reasonable sensitivity but less specificity. Measures in this category include specific indicator surfaces and DMFT increment in the previous year. Among the more useful specific tests have been mutans streptococci and lactobacillus counts and measurement of saliva buffering capacity. Other methods that show some promise include the physical measurement of incipient carious lesions of enamel. The measurement of possible determinants of a multifactorial disease is extremely difficult and regardless of which single method has been tried the authors have usually concluded that it is difficult to develop a reliable method of identifying caries susceptible individuals from the method. To a lesser extent the same conclusions have been applied to methods for identifying risk groups. There does, however, appear to have been little research in which a combination of tests or methods have been used, particularly combinations of tests that involve different scientific disciplines.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3053463 TI - Current regenerative periodontal therapy. AB - Healing, after subgingival curettage, leads to the formation of a long junctional epithelium. This is a repair process and not one of regeneration. Flap procedures of themselves also seem to lead to the formation of a long junctional epithelium, notwithstanding some evidence of bone repair. Bone grafts favour osseus repair without creating new attachment, for the previous architecture and function of the tissues is not restored. Recently, use of porous hydroxyapatite implants in block or granular form has led to pocket depth reduction and attachment gain. Conditioning of the root surface, after planing, with citric acid, attachment proteins or tetracycline has been widely investigated. The use of citric acid has not led to significant periodontal regeneration but the local use of slow-release tetracycline may prove to be of value. Guided regeneration aims to re-populate the root surface with active periodontal ligament cells. In recent experiments substantial reductions in pocket depth have been achieved. PMID- 3053465 TI - Comparative studies on the expression of tumor-associated glycoprotein (TAG-72), CA 19-9 and DU-PAN-2 in normal, benign and malignant pancreatic tissue. AB - Expression of tumor-associated glycoprotein (TAG-72) was examined by immunohistochemistry in pancreatic specimens from normal donors, and from patients with chronic pancreatitis and pancreatic cancer, and was compared with expression of CA 19-9 and DU-PAN-2. In the normal pancreas, TAG-72 was expressed in fewer ductal and ductular cells than were CA 19-9 (p less than 0.05) and DU PAN-2 (p less than 0.01 and 0.001 respectively), whereas in chronic pancreatitis all 3 antigens were expressed in ductal cells but only CA 19-9 and DU-PAN-2 in ductular cells (p less than 0.001). In the specimens from normal pancreas and chronic pancreatitis cases, TAG-72 was localized in the Golgi region, whereas CA 19-9 and DU-PAN-2 showed diffuse cytoplasmic and glycocalyx patterns. In pancreatic cancer, although the rate of expression of all 3 antigens was similar, their cellular localization differed significantly: only TAG-72 was expressed in the Golgi region (p less than 0.001), whereas CA 19-9 showed mainly glycocalyx (p less than 0.05) and/or intra-luminal patterns (p less than 0.01) compared with that of the other 2 antigens. We conclude that, due to differences in expression of TAG-72 in benign versus malignant cells, the monoclonal antibody against TAG 72 (B72.3) may be suitable for radio-immunodetection or radio-immunotherapy of pancreatic cancer. PMID- 3053466 TI - Melanoma antigen expression and metastatic ability of mutant B16 melanoma clones. AB - The biological functions of murine melanoma-associated antigens recognized by monoclonal antibodies (MAbs) (M562, M622 and M2590) were examined by using mutant clones which differed in their degree of expression of these antigens. Four clones of high expressors of 3 types of antigen (MEA group), 5 clones of low or non-expressors of M562- and M622-recognizing antigens (MEB group) and 4 clones of non-expressor of GM3 recognized by M2590 (MEC group) were used. Attachment of these clones to components of extracellular matrix was different between the groups. Two clones of the MEA group showed the highest ability to adhere to laminin and type-IV collagen, whereas the clones of the MEB and MEC groups significantly lost their ability to attach to laminin and type-IV collagen. In experimental lung metastasis, metastasizing ability of MEA-group cells was higher than that of MEB- and MEC-group cells. Our results suggest that these antigens play some functional role in metastasis mediated by increasing capacity for attachment to laminin and type-IV collagen. PMID- 3053468 TI - Utilization of human hematopoietic cell lines for the propagation and characterization of HBLV (human herpesvirus 6). AB - Details of the productive infection of established human cell lines of diverse origin by HBLV (also designated Human Herpesvirus 6) are described in this report. The infection and replication of HBLV in several T and B lymphoid and other cell lines was observed by electron microscopic examination, by the detection of viral antigen expression by indirect immunofluorescence assay (IFA) and by the presence of HBLV DNA by Southern blot hybridization. Several of these cell lines produced large amounts of virus. For this reason and because of the absence of other human herpesviruses, these lines have provided a valuable resource for the preparation of reagents and the development of assays for the detection and characterization of HBLV. The isolation and characterization of new HBLV isolates from patients with chronic fatigue syndrome were also facilitated by using some of the cell lines reported here. The host range of HBLV in established cell lines, therefore, does not appear to be limited to the B lymphocytes, as initially suggested by in vivo studies. The infection of T and B lymphocytes, megakaryocytes and neuronal cells in vitro suggests a need for the evaluation of diverse hematological and neurological disorders to shed light on a possible HBLV involvement. PMID- 3053467 TI - Molecular and cellular characterization of drug resistant hamster cell lines with alterations in ribonucleotide reductase. AB - Ribonucleotide reductase consists of 2 protein components frequently called M1 and M2. Hydroxyurea specifically inhibits DNA synthesis by interacting with the M2 protein and destroying a unique tyrosyl-free radical. We have carried out a molecular and cellular characterization of 2 Chinese hamster ovary cell lines exhibiting either low (HN(R)-AT) or relatively high (H(R)-R2T) resistance to the cytotoxic effects of hydroxyurea. Both drug-resistant lines have an increased level of ribonucleotide reductase activity. EPR measurements for tyrosyl-free radical content and studies with M1-specific antibodies indicated that the elevation in enzyme activity was entirely due to an increase in the M2 component. Studies with M1 cDNA showed that both drug-resistant cell lines contained a wild type level of M1 mRNA and a wild-type M1 gene copy number. Studies with M2 cDNA indicated that the 2 drug-resistant lines possessed elevated levels of M2 message that could explain the observed increase in M2 component. The elevation of M2 mRNA in the most resistant line, H(R)-R2T, was due to an increase in M2 gene copy number. The low resistant cell line, HN(R)-AT, exhibited a wild-type M2 gene copy number, indicating that the increase in M2 gene message occurred through a process other than gene amplification. Enzyme kinetic studies with partially purified preparations from both drug resistant lines showed reduced sensitivity to hydroxyurea and to the negative allosteric effector, dATP. In addition to hydroxyurea, H(R)-R2T cells were also resistant to several other drugs whose site of action is the M2 component. Furthermore, H(R)-R2T cells were not cross resistant to colchicine or puromycin, suggesting that hydroxyurea-resistant cells do not share the multi-drug resistance phenotype, which is frequently associated with cross-resistance to these drugs. PMID- 3053469 TI - Osteoarthritis of the wrist secondary to non-union of the scaphoid. AB - The surgical treatment of osteoarthritis of the wrist secondary to pseudarthrosis of the scaphoid is presented in the light of twenty years experience, and with improved knowledge of the pathological anatomy of this problem. The condition is difficult to treat owing to the varied pathology, the different needs of the patients and the alternative treatments available. Where a pseudarthrosis exists with involvement of the scaphoid-radius joint only, we recommend radial styloidectomy and repair of the pseudarthrosis. Where the pseudarthrosis is associated with carpal collapse which can be corrected, and osteoarthritis is confined to the joint between the capitate and lunate bones, we recommend correction of the carpal collapse and an arthrodesis between the lunate, capitate, hamate and triquetrum, with either anatomical repair of the scaphoid or eventual replacement with a Silastic implant. Where there is irredicable carpal collapse or lunate dislocation, proximal row carpectomy is recommended as an alternative to arthrodesis. Pseudarthrosis of the proximal pole of the scaphoid is best treated by replacement of the small bone fragment with a Silicone rubber implant. For pseudarthrosis with a painful arthritic joint of radius, scaphoid and lunate without involvement of the other periscaphoid joints, we recommend arthrodesis of the affected joints only. Finally, in cases of pseudarthrosis with advanced degenerative changes in the radiocarpal and intercarpal joints with associated collapse and deformity, or following unsuccessful resection, we recommend arthrodesis of the wrist. PMID- 3053470 TI - Non-toxic endotoxin polysaccharide induces soluble mediators which potentiate antibody production by murine retrovirus-suppressed splenocytes. AB - Mice infected with Friend leukemia virus show marked acquired immunodeficiency characterized by the impairment of immune function of spleen cells to various antigens, both in vivo and in vitro. The large mol. wt. endotoxin derived from Serratia marcescens, as well as a smaller non-toxic polysaccharide derivative, were found to augment the antibody responsiveness of spleen cells from normal as well as FLV-infected mice. In addition, serum from normal donor mice pretreated with BCG and injected either with endotoxin or the polysaccharide derivative potentiated the antibody response of spleen cells from both normal and FLV infected mice. Similar enhancement was induced by "antibody response helper factor(s)" present in 3-5 day spleen culture supernatants from endotoxin or polysaccharide-treated spleen cells from normal mice. Enhancement of the antibody response of spleen cells from FLV-infected mice by the antibody helper activity was due to stimulation of B-lymphocytes and reversal of a defect in antibody helper factor(s) formation by macrophages. Similar antibody response enhancing activity was induced by both endotoxin and the non-toxic polysaccharide derivative in cultures of normal spleen cells, adherent spleen cell populations, peritoneal cells and the P388D1 macrophage cell line. PMID- 3053471 TI - Low molecular weight immunopotentiators. AB - It has long been recognized that modulation of the immune system by various agents may have potential for the management of certain infectious and neoplastic diseases. Both natural products as well as chemically synthesized compounds have been investigated for immunotherapeutic potential. Over the years, conflicting reports on the clinical efficacy of these agents have left the early promise of immunotherapy unfulfilled. However, the manipulation of the immune system to generate a desired effect is becoming feasible as the mechanisms which regulate the immune network are better understood. Much of the early work on immunotherapy concentrated on the development of immunopotentiators, agents which enhance the host's own immune system against cancer cells or infectious pathogens. Furthermore, with the development of subunit and/or synthetic vaccines, which are often weakly immunogenic, the importance of developing agents capable of acting as adjuvants became apparent. As a result, the utility of immunopotentiators has now extended to the area of vaccines. There are a number of reviews available on immunomodulators [see Fenichel, R. L. and Chirigos, M. A. (eds) (1984), Immune Modulation Agents and Their Mechanisms, Marcel Dekker, New York]. The purpose of this article is to provide an update on low molecular weight agents capable of potentiating the immunological network. Attention will be given to those agents which have undergone significant clinical development in the areas of cancer, infectious diseases and vaccination over the past several years. These agents will be categorized as to whether they are naturally occurring or chemically synthesized. PMID- 3053472 TI - Effects of temperature and water immersion on plasma catecholamines and circulation. AB - The effects of environmental air temperature of 6 degrees C and 26 degrees C on catecholamines (CA) and circulation were studied in eight male subjects during rest and during bicycle exercise at WOBLA for 45 min each. We found that resting at 6 degrees C increased the norepinephrine (NE) levels to the same levels as endurance exercises at 6 degrees C. The increase of CA levels was 2.5 to 3 times higher during work at 26 degrees C compared with 6 degrees C. During both rest and exercise at 6 degrees C we found a higher stroke volume of the heart and a reduced heart rate (HR) with no or only small effects on the oxygen uptake and blood lactate levels compared with 26 degrees C. Measurements of the skin temperatures showed large differences both at rest and during work; those of core temperature showed no changes at rest and a slightly more pronounced increase during work at 26 degrees C compared with 6 degrees C. The behavior of CA, plasma renin activity (PRA), plasma aldosterone (PA), and circulation were studied in 13 top class swimmers and 12 recreational swimmers during immersion into water of 27 degrees C for 10 min. The recreational swimmers were additionally immersed into water of 21 degrees C and 33 degrees C. Even immersion at 33 degrees C induced a small but significant increase of NE levels and of blood pressure (BP) values with no effect on the HR and blood lactate values. Epinephrine (EPI) showed a tendency to decrease.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3053473 TI - Differences in sympathoadrenal, hormonal, and metabolic adaptation to submaximal and maximal arm and leg work compared with whole stroke in breast-style swimming. AB - During work on land adaptational reactions of circulation and hormones are influenced by the body position, the activated muscle mass, and the working extremities. The aim of this study was to explore the additional effect of water immersion during swimming on cardiocirculatory, metabolic, and hormonal regulation under different working conditions. Twelve young men not specifically trained in swimming underwent swimming tests on 3 different days. They had to swim breast stroke in total as well as isolated with legs or arms only each for 10 min at submaximal intensity and 150 m or 100 m at maximal intensity. This study was focused on changes of catecholamines (CA), heart rate (HR), blood pressure (BP), glucose, lactate, plasma renin activity (PRA), and plasma aldosterone (PA). Additionally, parameters of the electrolyte-volume homeostasis were investigated. Norepinephrine (NE) and epinephrine (EPI) increased during swimming under submaximal and maximal conditions. The augmentation of CA and HR was the highest after swimming whole stroke and the lowest after swimming arm stroke only. They were related to intensity within one type of swimming and showed a dependence on muscle mass independent from lactate or glucose levels when the different types of swimming were compared, although a positive correlation between CA and lactate levels was found. The BP was higher after leg work than after arm work, contrary to observations done on land regarding the comparable submaximal work loads. We suppose that this is an effect of water immersion and the horizontal body position whereby the central hemodynamic circulation becomes stabilized.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3053474 TI - Sympathoadrenergic regulation of metabolism and cardiocirculation during and following running exercises of different intensity and duration. AB - In three different field studies running loads of 2 X 200 m and 400 m, 3 X 1000 m and 3000 m, and finally 10,000 m were performed by respective groups of sprinters, middle-, and long-distance runners. We investigated the effects of exercise on the sympathoadrenal system by determining catecholamine (CA) concentrations in the venous blood and urine [free norepinephrine (NE), epinephrine (EPI)] and the respective sulfoconjugates in plasma and various hormonal, metabolic, and cardiocirculatory parameters. Endurance-trained athletes showed a lower heart rate (HR) and plasma renin activity (PRA) at rest in comparison with the sprinters. The concentrations of the plasma CA reflected the intensity more than the duration of exercise. At rest and following recovery the ratio of free NE/EPI in plasma was markedly higher in the group of sprinters in comparison with the long-distance runners. During exercise, however, an opposite movement occurred resulting in a higher ratio in the latter group. The pre-start ratios of NE/EPI in urine were similar to those in plasma. The sports disciplines' specific differences in NE/EPI both at rest and during exercise suggest that the overall sympathetic activity, reflected by NE and EPI, is regulated in a quite differentiated pattern. The sulfoconjugated CA, however, increased less clearly after the middle- and long-distance bouts than the respective free CA which caused the ratio sulfates/free CA to decline. There were strong relations between the levels of free plasma NE and EPI and that of blood lactate which, however, rather reflects a parallelism subsequent to the highly intensive stimulation. PMID- 3053475 TI - Sympathoadrenergic regulation in elite fencers in training and competition. AB - In ten fencers at the top national level the adrenergic regulation was investigated by determining the free and sulfoconjugated catecholamines (CA) in a training fight and a competition for the national championship to evaluate the influence of emotional strain in this discipline. In the training fight which is physically more strenuous, norepinephrine (NE) was significantly higher (+27%) than in the championship contest after which the epinephrine (EPI) level was more increased (+76.5%). Accordingly, the systolic blood pressure (SBP) was higher during competition and correlated with EPI. The share of the sulfated CA in their total amount was reduced in both loads. Since energy production in fencing is predominantly alactacitic during maximal loads of short duration and aerobic during submaximal load intensities, the lactate level in the training fight was below the anaerobic/aerobic threshold. The significantly higher lactate, glucose, and alanine levels during the fight for the national championship, presumably induced by the additional stimulation of the anaerobic muscular and hepatic glycogenolysis as well as muscular glycolysis, may be accounted to the fortified EPI secretion caused by emotional strain. The higher cortisol and renin levels may be explained by the strong central stimulation and the direct peripheral effect of EPI respectively. PMID- 3053476 TI - Cardiocirculatory, hormonal, and metabolic reactions to various forms of ergometric tests. AB - The sympathoadrenergic reaction is not only dependent on the duration and intensity of work but also on the body position and the involvement of small or large muscle masses. This observation made in field tests comparing different sports disciplines such as swimming, running, or diving encouraged us to investigate this topic under the following laboratory conditions. Twelve healthy sport students participated in ergometric tests on a bicycle ergometer in a horizontal and vertical body position as well as on a treadmill and a swim bench ergometer. The changes of plasma catecholamines (CA) obtained in the different ergometric tests were compared with those cardiocirculatory, metabolic, and hormonal parameters which can be influenced by the sympathoadrenergic stimulation. In the horizontal body position we found a smaller increase of norepinephrine at submaximal and maximal work loads combined with a similar reaction of renin, whereas the diastolic blood pressure and the mean arterial blood pressure increased more. The substrates of lipolysis and aerobic and anaerobic glycolysis did not show obvious differences depending on the body position. In the swim bench test, however, the lactate increase started earlier and was comparatively higher than in the other ergometric tests in which the maximal work load and VO2max were higher. Although a smaller muscle mass was used and a lower maximal oxygen uptake was reached, we did not find statistically different CA values during the swim bench ergometric test compared with the bicycle ergometric test in a horizontal body position. In our ergometric tests, the venous CA levels (especially norepinephrine) were predominantly influenced by the body position. PMID- 3053477 TI - Hypoglycemia and aggression: a review. AB - The popular notion that a tendency to develop low levels of blood glucose is the cause of a range of behavioral problems is reviewed. It is concluded that it is inappropriate to use the glucose tolerance test as a test of the tendency to develop reactive hypoglycemia. Instead, a meal tolerance test, in which glucose is administered in the presence of fat and protein, should be the method of choice. The use of a meal tolerance test strongly suggests that reactive hypoglycemia rarely results, except in a few exceptional individuals. Three situations are described in which a correlation between a tendency to develop moderately low levels of blood glucose during a glucose tolerance test (not hypoglycemic values) and the tendency to act aggressively have been reported. The significance of these data is unclear but several possible mechanisms by which glucose may influence behavior are discussed. PMID- 3053478 TI - A reexamination of the hypoglycemia-aggression hypothesis in laboratory mice. AB - The suggestion of a clinical link between hypoglycemia and aggression has been recently encouraged by the work of Virkkunen and others. Modern techniques have impressively established a correlation between reactive hypoglycemia (low blood sugar levels engendered by stimulation of endogenous insulin secretion) and hostility as expressed in a variety of situations. It is not surprising that blood sugar levels influence behavior, as the brain is entirely dependent on glucose reaching it via the vascular system. In spite of this impressive link, there are remarkably few successful studies involving the effects of manipulating glucose levels on fighting in infrahuman animals. As there have been several methodological developments in this area it was thought appropriate to reinvestigate such phenomena in mice. The effects of injected bovine insulin and glucose were assessed using an ethopharmacological methodology applied to social encounters by isolated male Swiss mice with docile anosmic opponents. Although manipulation of blood glucose levels did not consistently change attack or defensive behavior, these treatments did modify aspects of the interactions. PMID- 3053479 TI - Dominance and aggression. AB - Dominance is a complex phenomenon mediated by different mechanisms. Various motivations and their mutual correlations determine the tendency to dominate. A subject that is dominant in every situation (i.e., absolutely dominant) is rather an exceptional case. Dominance may be limited to particular situations and exhibited only with some definite partners. One subject may be dominant over one partner and submissive with another. Aggressive behavior is not indispensable to obtain and keep dominance status. It seems that dominance sustained without aggression is more stable than dominance formed on the basis of aggressive display, since experiments on predatory dominance in pairs and groups of cats support such an assumption. Various brain structures were found which are involved in aggressive behavior, but in respect to dominance our experiments point to the role of the dorsal amygdala in predatory dominance. PMID- 3053480 TI - Potential and limitations of the offense-defense dichotomy in aggression research, considering the complex and evolving interactions of biological, psychological and social determinants of aggression. AB - The offense-defense dichotomy undeniably proved to be of heuristic interest in that it allowed relevant distinctions between specific behaviour patterns, to establish correlations between situations in which either "offensive" or "defensive" patterns are likely to be displayed, and to uncover differential drug effects on those distinct behavior patterns. But for a number of reasons, the author is led to conclude that even though the offense-defense dichotomy indeed helps in many instances to describe and to distinguish, it has rather limited potential really to explain in depth what it distinguishes at a phenomenological level of investigation. PMID- 3053481 TI - Adaptive pain inhibition in murine resident-intruder interactions. AB - In dyadic encounters with aggressive resident conspecifics, male intruder mice display an initial acute nonopioid analgesia followed by a more enduring opioid analgesia. The former reaction occurs in association with active defense (flight or fight) and can be seen in response to the scent of an aggressive conspecific or defeat experience per se. In contrast, the latter (opioid) reaction is associated with passive defense (immobility) and occurs in response to extended conspecific attack. The mechanisms underlying these two ecologically-relevant forms of pain inhibition are contrasted and the phenomena are discussed in relation to the question of adaptive significance. PMID- 3053482 TI - Recent research into Behcet's disease in Japan. AB - Recent studies on Behcet's disease supervised by a national research committee are reported under the headings of epidemiology, the role of streptococcus infection, PMN activity, immunological changes and treatment, concluded with a revised set of diagnostic criteria. PMID- 3053483 TI - Florence Nightingale: statistician and consultant epidemiologist. AB - Florence Nightingale became a legend in her own lifetime and the image of the lady with the lamp still influences public thinking about the practice of nursing and its practitioners. Like many legends, her story has been adapted to suit changing circumstances and purposes. One could speculate why it is that particular aspects have come into greater prominence at particular times in history. This article explores a less-known aspect of her work, her expertise and use of statistics and epidemiology. And it illustrates how that expertise was called upon to address a major health problem in New Zealand in 1860. PMID- 3053484 TI - Immunological mechanisms of allergic disease. PMID- 3053485 TI - Ocular allergy. Mast cells and eosinophils. PMID- 3053486 TI - Differential diagnosis of ocular allergy. PMID- 3053487 TI - Evaluation and treatment of the allergic patient. PMID- 3053488 TI - Allergic conjunctivitis. PMID- 3053489 TI - Vernal keratoconjunctivitis. PMID- 3053490 TI - Giant papillary conjunctivitis. PMID- 3053491 TI - Contact allergy and toxicity in the eye. PMID- 3053492 TI - Chronic allergic conjunctivitis. PMID- 3053493 TI - Laboratory evaluation of ocular allergy. PMID- 3053494 TI - The complement system in ocular allergy. PMID- 3053495 TI - New directions in therapy for ocular allergy. PMID- 3053496 TI - Conjunctival provocation tests and naturally occurring allergic conjunctivitis in clinical trials. PMID- 3053497 TI - Transferrin and the growth-promoting effect of nerves. AB - In addition to its role in the activity of specialized proteins such as hemoglobin and myoglobin, iron is required as a cofactor in several important enzymes common to most animal cells. One such enzyme, ribonucleotide reductase, which regulates the production of deoxyribonucleotides during DNA synthesis, requires a continuous supply of iron to maintain its activity throughout the process of DNA replication. The mechanism by which animal cells normally acquire iron involves receptor-mediated uptake of iron-loaded transferrin, followed by release of apotransferrin. The density of transferrin receptors on the cell surface is greatly increased in rapidly dividing normal and neoplastic cells. Various mitogens and certain organogenic tissue interactions have been shown to induce the appearance of transferrin receptors, signalling the onset of DNA replication. Interference with this process of iron delivery causes the rapid arrest of cell cycling, frequently during the S phase itself, which underscores the importance of iron for DNA replication. Although most circulating transferrin is synthesized in the liver and embryonic yolk sac, smaller quantities are produced in several other embryonic organs and certain other adult tissues. It has been suggested that local synthesis and/or release of transferrin supplies the iron required by rapidly growing cells in situations where the cells do not have ready access to adequate amounts of plasma transferrin due to incomplete development of the vasculature or the presence of blood-tissue barriers (Ekblom and Thesleff, 1985; Meek and Adamson, 1985). Oligodendrocytes and Schwann cells have been shown to synthesize and/or contain high concentrations of transferrin and these cells therefore may constitute a local source of this factor for neurons, whose growth and survival in vitro require transferrin. Transferrin in central and peripheral nervous tissues may be significant for the trophic or growth-promoting effect neurons exert on cells of certain tissues. Transferrin duplicates the activity of neural tissue or neural extracts on growth and development of cultured skeletal myoblasts from chick embryos and on proliferation of mesenchymal cells in blastemas from regenerating amphibian limbs, two systems that have been widely used in investigations of the growth promoting influence of nerves. Moreover, removal of active transferrin from neural extracts, either with antibodies to transferrin or chelation of the iron, inhibits reversibly the effect of the extract in these developing systems. While the physiological significance of the extract in these developing systems.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3053499 TI - Problems of immune surveillance over the cytodifferentiation state and some cellular mechanisms of natural antitumor resistance. PMID- 3053498 TI - Protein secretions of Sertoli cells. AB - As was stated in the introduction, many of the functions of the Sertoli cells are apparently carried out by the protein secretions of these cells. The use of Sertoli cell cultures and appropriate biochemical and immunological techniques has allowed the characterization of some of these secretion products. It is likely that many of the functions of the Sertoli cells are necessary because of the presence of the blood-testis barrier. Many growth and nutritive factors which are necessary for cell viability are available to most cells via the serum. The germinal cells within the adluminal compartment do not have access to serum factors and one of the functions of the Sertoli cells is to synthesize serum-like components and secrete them into the adluminal compartment. The historical description of Sertoli cells as "nurse cells" thus appears to have been accurate. The nurse-cell function is most clearly demonstrated by the proposed mechanism by which germinal cells obtain ferric ions. The Sertoli cells have developed a system to move serum-derived iron through their own cytoplasm and to secrete it bound to newly synthesized testicular transferrin molecules which can deliver it to specific receptors on the germinal cell surface (Huggenvik et al., 1984). Functionally, all of the secreted proteins from Sertoli cells which have been characterized or proposed fall into one of five basic classes. First, Sertoli cells secrete a number of transport proteins including transferrin, ceruloplasmin, and ABP. The proposed function of these proteins is the transport of Fe3+, Cu2+, and androgens to the germinal cells or to the epididymis (ABP). Second, Sertoli cells synthesize and secrete a number of proteins which have a hormone-like or growth factor-like activity. AMH is a clear and well-documented example of this type of product while the evidence for inhibin, somatomedin C, EGF-like growth factor, and seminiferous growth factor will require further corroboration. Third, Sertoli cells secrete proteins which have enzymatic activities. Plasminogen activator is the best characterized example of this class of products and the alpha-lactalbumin-like activity is of potential interest. The fourth class of Sertoli cell secretion products includes those proteins which contribute to the basement membrane, namely, type IV collagen and laminin. Finally, there is a very important group of Sertoli cell secretion products for which there is, as yet, no evidence for a defined function. This group includes SGP-1 and SGP-2 which are the major sertoli cell products in rats and which have been well-characterized biochemically.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3053500 TI - Nuclear function and organization: the potential of immunochemical approaches. PMID- 3053501 TI - [AIDS: an interim statement by the WMA]. PMID- 3053502 TI - [Symptoms, which indicate thyroid disease]. PMID- 3053503 TI - [Diagnostic strategies: determination of thyroid gland hormones in serum]. PMID- 3053504 TI - [The significance of microsomal antibodies, thyroglobulin antibodies, thyrotropin receptor antibodies in the diagnosis of thyroid diseases]. PMID- 3053505 TI - [Scintigraphy of the thyroid gland]. PMID- 3053506 TI - [Sonography and controlled fine needle puncture of the thyroid gland]. PMID- 3053507 TI - [Therapy of endemic struma]. PMID- 3053508 TI - [Therapeutic strategies in immunogenic hyperthyroidism and endocrine ophthalmopathy]. PMID- 3053509 TI - [Therapy of hypothyroidism]. PMID- 3053510 TI - [Thyroid cancers--current diagnostic and therapeutic strategies]. PMID- 3053511 TI - [Surgical indications and choice of surgical procedure in thyroid gland diseases]. PMID- 3053512 TI - [Therapeutic strategies--nuclear medicine aspects]. PMID- 3053513 TI - [Thyroid gland diseases: long-term treatment and follow-up]. PMID- 3053516 TI - [Bone pain of arterial origin]. PMID- 3053514 TI - [Hemobilia following erosion of the gastroduodenal artery and common bile duct by a pancreatic cyst]. PMID- 3053515 TI - [Spontaneous cholecysto-gastric fistula]. PMID- 3053517 TI - [Drug therapy of hypertension in pregnancy]. PMID- 3053518 TI - [Kidney diseases in pregnancy]. PMID- 3053519 TI - [Diabetes and pregnancy]. PMID- 3053520 TI - [Acquired coagulation disorders in obstetrics]. PMID- 3053521 TI - [Oncologic problems in pregnancy]. PMID- 3053522 TI - [Drugs and pregnancy]. PMID- 3053523 TI - [Recurrence of a central Cushing's syndrome following transsphenoidal surgery of the pituitary]. PMID- 3053525 TI - [Splenomegaly and peripheral pancytopenia in a 62-year-old patient]. PMID- 3053524 TI - [A 39-year-old heavy smoker with acute effort dyspnea and fever]. PMID- 3053526 TI - [A primary skin tumor in myeloproliferative disease]. PMID- 3053527 TI - [A 61-year-old patient with a 15-month history of progressive diffuse abdominal pains]. PMID- 3053528 TI - The effect of tissue dose and site of grafting on immunity to corneal allografts. AB - The purpose of this investigation was to determine the quantitative relationship between corneal alloantigens and host immunity. In addition, the effect of the site of introduction of the corneal antigens on the host response was determined. Two alloantigenic strains of rats were reciprocally grafted at three different locations in the body with carefully quantitated amounts of corneal tissue: (1) orthotopically in the cornea of the eye; (2) subcutaneously; and (3) intraperitoneally. Corneal tissue placed orthotopically into vascularized graft beds did not elicit a systemic immune response. Subcutaneous grafts elicited a weak systemic alloantigenic response, whereas corneal tissue placed in the peritoneal cavity consistently induced a vigorous cellular and humoral alloantigenic response. Eight or more full thickness corneal allografts grafted subcutaneously were required to elicit a systemic response. On the other hand, as few as four corneal allografts placed intraperitoneally invariably elicited a systemic alloimmune response. The results of this investigation demonstrate that both the amount and route of introduction of alloantigen affect the recipient's response to corneal tissue and that the rejection of a single orthotopic cornea graft is a site-limited response. Immune effector cells were not found in the spleens and alloantibodies were not present in the serum of animals that had rejected corneal allografts. PMID- 3053530 TI - Ocular renin-angiotensin: immunohistochemical evidence for the presence of prorenin in eye tissue. AB - Angiotensin II (A2) is a vasoconstrictor generated by the renin-angiotensin system. A2 appears to act also as an angiogenic factor. Recent evidence suggests that renin is synthesized at many tissue sites and may generate A2 locally. Local A2 may have important functions in the normal and diseased eye. We examined eight human eyes by immunostaining with an antibody to prorenin, the biosynthetic precursor of renin. In all eyes, prorenin staining was extensive in the pars plicata of the ciliary body suggesting that the ciliary body synthesizes renin and this renin may be part of an ocular A2 generating system. PMID- 3053529 TI - The use of bioerodible polymers and 5-fluorouracil in glaucoma filtration surgery. AB - A study was performed to examine the effect of a localized and sustained delivery of 5-fluorouracil (5-FU) on the success of glaucoma filtration surgery in 18 rabbits in a prospective, randomized, double-masked and placebo-controlled fashion. A bioerodible polyanhydride composed of bis (p-carboxyphenoxy) propane and sebacic acid was used as the drug carrier. The polymer and 5-FU (20% by weight) were compressed into 3 mm diameter discs, 1 mm thick. The polymer with the 5-FU was randomized to one eye and the fellow eye received the blank polymer. The results showed that intraocular pressures (IOP) were lower in the experimental eyes during the 5th through 17th postoperative days, but eventually both experimental and control eyes returned to preoperative levels. Filtration blebs lasted longer in experimental eyes when compared to control eyes. Implant disappearance occurred after IOP elevations and bleb failure. Eventually, the filtration surgery failed in both the experimental and control rabbit eyes. PMID- 3053531 TI - [Clinical aspects and genetics of congenital skin defects]. AB - Congenital skin defects are uncommon anomalies in neonates, confronting the clinician with diagnostic problems because of their etiologic heterogeneity. The classifications currently in use are didactically unsatisfactory. Therefore, we propose a new classification taking account of the localization of the congenital skin defect (head, trunk, extremities) and the criterion of isolated or associated occurrence. The groups clinically defined in this way, tables showing the pathognomonic symptoms and, in addition, a short review of all syndromes involving congenital skin defects should allow a rapid diagnosis. Clinical and genetic features of the isolated congenital skin defects are also reviewed. PMID- 3053532 TI - [Microbial flora and odor of the healthy human skin]. AB - The microflora resident on human skin shows great interindividual and intraindividual differences. It is essentially composed of micrococci, staphylococci, aerobic and anaerobic coryneforms as well as pityrosporum species which, in accordance with the different environment in the different regions of the body, are in a steady state. With increasing age, human skin microflora undergoes qualitative changes: the streptococci, which are found in infants, disappear and coryneform bacteria occur, which are mainly responsible for odor production. Anaerobic propionibacteria are more numerous in juveniles and young adults, a fact that may be explained by increased sebum production. Only the coryneform bacteria are able to produce the typical axillary odor by decomposition of apocrine sweat. Cocci, however, obviously do not have this capacity. It remains to be established which substances participate in odor production. With sensitive chromatographic methods amino acids, steroids and free fatty acids were detected, which could be related to body odor. There are possibly only a few commonly occurring odorous substances. The necessity of analyzing these substances is stressed. PMID- 3053534 TI - [Edmund Lesser and his (forgotten) school]. PMID- 3053533 TI - [Fine needle aspiration of the skin: diagnosis of malignant lymphoma and of cutaneous manifestations of myeloid hemoblastosis]. AB - Fine-needle aspiration cytology of the cutis was carried out in 103 patients suffering from malignant lymphomas and myelogenous leukemias. All biopsies were performed using a novel aspiration device. Only in 7.8% of cases was it not possible to establish a cytological diagnosis because the biopsy specimens obtained were inadequate. The sensitivity of the method was 96.8%. There was a close correlation between the cytological and the final diagnosis (contingency coefficient C = 0.96). The method was equally efficient for primary evaluation as it was for the follow-up of patients with hematological neoplasms. PMID- 3053536 TI - Stress and relaxation in health visiting. PMID- 3053535 TI - [Gingival hyperplasia and serous papules due to cyclosporin treatment]. AB - After 3 months of immunosuppressive treatment with cyclosporin because of a focal segmental glomerulosclerosis, a 13-year-old female developed marked gingival hyperplasia, hypertrichosis of the arms and papulo-vesicular skin lesions on the left leg. Histological and immunohistological examinations of the gingivae showed hyperplasia of the mucosa with perivascular infiltrates of plasma and B cells in the submucosa. The lesions on the leg revealed signs of cutaneous vascular inflammation with eosinophils, but there was no evidence of leucocytoclastic vasculitis. Ten months after discontinuation of cyclosporin, the lesions had nearly regressed. The patient's renal function deteriorated progressively to the point of terminal insufficiency. PMID- 3053537 TI - Hypoxic toxicity of misonidazole in a glucose-6-phosphate dehydrogenase deficient mutant Chinese hamster ovary cell line. AB - The metabolic activation of misonidazole (MISO) and its effects on the hexose monophosphate pathway (HMP) and on cell viability were studied in hypoxic mutant Chinese hamster ovary (CHO) cells deficient in glucose-6-phosphate dehydrogenase and their parent wildtype cells. The metabolic activation of MISO was similar in both cell lines as indicated by the binding of 14C-MISO to the acid-insoluble fraction of these cells; it was decreased by the absence of glucose. In the wildtype CHO cells, MISO caused a significant stimulation of the activity of the HMP while in the mutant CHO cells no HMP activity was measurable, even in the presence of MISO. In both cell lines clonogenicity began to decline after 2 hr and trypan blue exclusion after 4 hr of hypoxic incubation. The effect of MISO on both parameters of cell viability was somewhat more pronounced in the wildtype CHO cells. This difference became especially significant at the longer incubation times. The results indicate that reducing equivalents for the metabolic activation of MISO are provided not only by the HMP but that pathways other than the HMP, such as glycolysis or pathways starting from mitochondrial tricarboxylates, are of similar or even greater importance in this respect. PMID- 3053538 TI - Design and characteristics of a facility for total-body and large-field irradiation. AB - A facility for total-body X ray irradiation has been built using two 4 MV linear accelerators, one supported from the ceiling and one placed in a floor pit. The maximum distance between the sources is 410 cm. The patient lies supine on a light, movable support with a stretched-canvas top, halfway between the sources where the field size is 80 X 220 cm2. A special flattening filter maintains the doserate variation in air within +/- 3% over the central 70 X 200 cm2 of the field, but some quality variations within the beam are noticeable. The doserate at 205 cm distance from the sources is variable between 0.05 and 0.8 Gy/min (half from each source). To permit treatment of large fields at higher doserates, the accelerators can be moved vertically to place the sources at 120 cm or 160 cm from the patient's midplane. For this purpose, independently movable collimators are provided and the flattening filter is designed to provide two options, one for the large total-body field and the other with less filtration covering a smaller solid angle. At 120 cm distance, each beam can provide a doserate of up to 1.1 Gy/min. PMID- 3053540 TI - The role of irradiation in solid organ transplantation. PMID- 3053539 TI - Important prognostic factors influencing outcome of combined radiation and hyperthermia. AB - Clinical experience with combined local-regional hyperthermia and radiation therapy has been rapidly accumulating over the past few decades. Its superior efficacy to the use of radiation alone has been demonstrated in several retrospective and prospective reports in the literature. It is evident now that there are several important factors that will influence the final outcome of the treated patients. The parameters that will be discussed in this paper include: I. Pretreatments factors: 1. tumor dimension 2. tumor histology 3. tumor site. II. Treatment factors: 1. radiation therapy dose 2. hyperthermia parameters: (a) thermal variables (b) number of heat treatments (c) sequence of hyperthermia and radiation treatments (d) hyperthermic device. Finally, evaluation of response and complications will also be discussed. The importance of abiding by an accepted reporting system will be emphasized, and clarification of times at which response assessments were made will also be discussed. With the availability of longer term follow-up, use of an actuarial method of reporting becomes more important. The future of hyperthermia and radiation remains very promising but a lot of questions still need to be answered by well-conducted and reported clinical trials. PMID- 3053541 TI - Gustave Roussy. PMID- 3053543 TI - Psychosocial context of adolescent development. Study group report. PMID- 3053544 TI - Health futures of adolescents. Bibliography of journal articles 1980-1987. PMID- 3053542 TI - Primary orbital lymphomas. AB - Between 1950 and 1982, seventeen patients with primary orbital lymphoma were treated at the University of Kansas Medical Center. There were 10 males and 7 females with a median age of 61 years. Four patients had bilateral disease, seven patients had disease involving the conjunctiva, and in ten patients, the disease involved paraocular structures. Fourteen patients received radiation with a median dose of 3500 cGy (range 2250 cGy to 4250 cGy) given in about 3 1/2 to 5 weeks. Median follow-up was 10 years (range 5 to 31 years). Local control was 100% and 5-year survival was 76%. Three patients are living with no evidence of lymphoma; three patients died from progression of the disease, and others died from unrelated causes. Radiation treatment for localized primary orbital lymphomas appears to be the primary treatment of choice. PMID- 3053545 TI - From Minnesota to Virginia--with many sojourns between. PMID- 3053546 TI - Cat scratch disease. PMID- 3053548 TI - Hydramnios causing uterine rupture in a mare. AB - An 18-year-old mare, 285 days pregnant, was evaluated for apparent abdominal pain of 8 hours' duration. A large volume of sanguinous fluid was obtained on abdominocentesis, and digital vaginal examination revealed a dilated cervix and blood in the uterus. Abdominal palpation per rectum revealed the uterus to be large and distended with fluid. Ultrasonography revealed a dead fetus on the floor of the cranial portion of the abdomen. The mare was euthanatized, and necropsy confirmed that the uterus had ruptured, and that the fetus, within its chorioallantois, was in the abdomen. The amniotic sac contained approximately 96 L of amniotic fluid. Torsion of the amniotic sac separated the fetus from the fluid-filled compartment. Hydramnios was diagnosed on the basis of the excessive amniotic fluid and was believed to be the cause of the uterine rupture. PMID- 3053549 TI - Doppler examination in varicocele. A standard method of evaluation. AB - Doppler sonography is considered a reliable method for detecting reflux due to venous valvular incompetence, which occurs in varicocele. It is, however, a matter of debate whether the characteristics of this reflux can be correlated quantitatively in a clinical setting. In this study, a two-step method was utilized to identify reflux in basal conditions with the patient standing and breathing spontaneously and to determine the time of centrifugal secondary venous reflux of the distal spermatic cord during the squeezing and relaxing maneuver. This method is closely related to that used by the vascular surgeon to detect valvular incontinence of the saphenous vein. Since Doppler sonography is much more reproducible than Valsalva's maneuver, it is therefore much more reliable. In a 12-month period, 625 patients with pathologic findings in at least two spermiograms were studied. Thirty percent showed constant basal reflux not influenced by respiratory exhilation. The squeezing and relaxing maneuver induced secondary reflux longer than 1.6 seconds in 17%, between 0.8 and 1.6 seconds in 17% and less than 0.8 seconds in 36% of the patients. The higher sensitivity and specificity of Doppler examination compared with thermography and angiography, as well as its low cost and noninvasiveness, make this the procedure of choice in the diagnosis of venous reflux in varicocele. PMID- 3053547 TI - Prevalence of udder infections and mastitis in 50 California dairy herds. AB - The California mastitis test (CMT) and bacteriologic culture were performed on samples of bulk-tank milk and cow-composite milk (n = 23,138 cows) from 50 California dairies, 19 of the 50 with known mastitis problems. Thirty-eight (76.0%) bulk-tank milk samples and 12,334 (53.3%) cows were positive by results of the CMT. Potential mastitis agents were isolated from 5,085 (22%) cows. Staphylococcus aureus was isolated from all 50 herds, Streptococcus agalactiae was isolated from 47 herds, and Mycoplasma sp was isolated from 24 herds. For cow composite milk samples, the prevalences were 9.3% for Str agalactiae, 9.1% for S aureus, 0.9% for Mycoplasma sp, 1.2% for coliform bacteria, 0.9% for other streptococci, 0.8% for coagulase-negative staphylococci, and 1.3% for other organisms. The relative sensitivity and the relative specificity of the CMT performed on cow-composite milk samples were 83.4% and 55.2%, respectively, and the predictive value of positive CMT results was 34.2%. PMID- 3053551 TI - Cefuroxime axetil. PMID- 3053550 TI - A40926 aglycone and pseudoaglycones: preparation and biological activity. AB - A40926 antibiotics are new glycopeptides which are much more active than other members of this class against Neisseria gonorrhoeae. Their activity against Gram positive bacteria, including coagulase-negative Staphylococci, is similar to that of other glycopeptides. An A40926 preparation containing factors A and B ("A40926 A + B complex") was hydrolyzed to the aglycone and to the mannosyl and N acylaminoglucuronyl aglycones. The mannosyl aglycone and the aglycone were less active than A40926 A + B complex against Streptococci and Gram-positive anaerobes and lost the anti-gonorrheal activity. In contrast, the N-acylaminoglucuronyl aglycones were as active as the parent complex against these Gram-positive bacteria and were moderately active against N. gonorrhoeae. The aglycone and, even more so, the N-acylaminoglucuronyl aglycones, had better activity than the parent complex against coagulase-negative Staphylococci. In experimental septicemia in the mouse, A40926 A + B complex and its derivatives had activity proportional to their MIC for the test organism. PMID- 3053552 TI - Drug transfer into cerebrospinal fluid after simultaneous administration of ampicillin with ceftriaxone or ceftazidime in rabbits with Staphylococcus aureus meningitis. AB - Meningitis was induced in white rabbits with a non beta-lactamase producing strain of Staphylococcus aureus. There was no significant difference in the CSF level of ceftazidime after simultaneous administration of ceftazidime with ampicillin or after ceftazidime alone. In contrast the percentage CSF penetration of ampicillin decreased from 8.81% after administration of ampicillin alone to 2.77% after the simultaneous administration of ampicillin with ceftazidime. The ratio of AUC in CSF and serum was 19.4% after ampicillin alone and 7.71% after simultaneous administration with ceftazidime. When ceftriaxone was combined with ampicillin there was no effect on the CSF penetration of either drug. Cephalosporins have unpredictable effects on the CSF concentration of ampicillin, probably due to variable effects on the penetration of ampicillin from blood to CSF. PMID- 3053553 TI - Penetration of ciprofloxacin and ofloxacin into human allograft pancreatic juice. AB - The penetration of ciprofloxacin (500 mg) and ofloxacin (400 mg) into pancreatic juice following a single oral dose, was investigated in seven patients who had undergone pancreatic transplantation. With a special technique for segmental pancreatic transplantation it was possible to collect pure pancreatic juice at regular intervals for up to 24 h after drug intake. The antibiotic concentrations were determined by the agar-well diffusion method. The concentration of ciprofloxacin in pancreatic juice was 36% of that in serum. The same figure for ofloxacin was 92%. The mean peak level in pancreatic juice was 0.5 +/- 0.0 mg/l (+/- S.E.) for ciprofloxacin and occurred at 4 h after drug intake. The same figure for ofloxacin was 2.7 +/- 0.7 mg/l (+/- S.E.) occurring at 3.6 h. The decrease in concentrations with time was parallel to the serum concentration curves for both antibiotics. The concentrations of ofloxacin in pancreatic juice exceeded the MIC values for most of the organisms causing infections in the pancreas for several hours after an oral dose. The concentrations of ciprofloxacin only briefly exceeded the MIC for the same organisms. PMID- 3053554 TI - Therapy of acute and chronic gram-negative osteomyelitis with ciprofloxacin. Report from a Swedish Study Group. AB - Oral ciprofloxacin was used at doses ranging from 500 mg to 1500 mg twice daily for 15 to 476 (mean 139) days for treatment of acute or chronic osteomyelitis in 38 patients, and acute arthritis in two. Clinical efficacy could be evaluated in 34 patients; 22 had resolution of their osteomyelitis, five improved and there were seven failures. Pseudomonas aeruginosa was the causative agent in 28 patients. It was eradicated in 22 patients, persisted but remained sensitive to ciprofloxacin in three and persisted with emergence of resistance to ciprofloxacin in three. Nineteen other pathogens, five Gram-negative and 14 Gram positive, were isolated. Of those, one strain of Staphylococcus aureus, two of Staph. epidermidis and three of Streptococcus faecalis remained sensitive to ciprofloxacin during treatment. In one patient, Slr. faecalis persisted with emergence of resistance to ciprofloxacin. Ten adverse events related to ciprofloxacin treatment were observed in nine patients; two phototoxic reactions, two cases of impaired colour vision, and one each of exanthema, abdominal pain, malaise, drug fever, peripheral neuropathy and eosinophilia. In three patients the adverse events led to treatment discontinuation. In conclusion, ciprofloxacin seems to offer an oral alternative to injectible antibiotics in patients with osteomyelitis caused by Gram-negative bacteria, including Ps. aeruginosa. PMID- 3053557 TI - Enhancement of antimicrobial activity of metronidazole in mixed cultures of Bacteroides fragilis and Escherichia coli. PMID- 3053556 TI - Selection of resistance to gentamicin and netilmicin in the faecal flora following prophylaxis for colo-rectal surgery. AB - The selection of aminoglycoside-resistant bowel flora, following the administration of either gentamicin or netilmicin in combination with metronidazole for prophylaxis, during colo-rectal surgery in 88 patients has been examined. Both antibiotic regimens resulted in the selection of an aminoglycoside resistant flora in a total of 57 (65%) of patients: in half of the patients there was a net gain in the aminoglycoside-resistant flora, and in 13 (15%) one aminoglycoside-resistant strain present prior to prophylaxis was displaced by another following operation. Three patients (3%) lost aminoglycoside-resistant strains after prophylaxis. Most of the resistant organisms selected were considered to be of little importance as potential pathogens, at least in the short term. In only a small minority (5%) of patients were aminoglycoside resistant enterobacteria isolated. Aminoglycoside-resistant Staphylococcus aureus was not isolated. Of the resistant enterobacteria, only one strain, an isolate of Enterobacter cloacae selected in a patient receiving gentamicin, carried a resistance determinant which was self-transmissible to Escherichia coli. PMID- 3053555 TI - A randomized trial of empirical antibiotic therapy with one of four beta-lactam antibiotics in combination with netilmicin in febrile neutropenic patients. AB - Over a two year period 174 evaluable episodes of fever in neutropenic patients were treated in a randomized study comparing four beta-lactam antibiotics, each given in combination with netilmicin. Exclusions included episodes due to viral or fungal infection, and trial violations. Most patients were receiving treatment for leukaemia, including 18% undergoing bone marrow transplantation. The overall response rate (EORTC criteria) was 66%, ranging from 56% for cefoperazone to 76% for mezlocillin. Microbial documentation was obtained in 31% of episodes; Gram positive isolates were most frequent but Pseudomonas aeruginosa was found in 18 patients. In patients with microbiologically documented infection 70% improved, overall--from 40% with cefoperazone to 80% with piperacillin (P less than 0.05). Nephrotoxicity was seen in 6.7% and was associated with severe documented sepsis. Hypokalaemia was seen in 29% and was most marked in patients receiving ticarcillin. Rashes occurred in 6.6% overall, with no difference between the groups. Ototoxicity, shown by serial audiograms, was seen in 4.7% of patients. No evidence of vestibular dysfunction was seen in 62 patients studied. Of thirteen deaths due to the primary infection, seven were caused by Ps. aeruginosa and five by fungi. PMID- 3053558 TI - Towards a fundamental understanding of the MIC of beta-lactam antibiotics. PMID- 3053559 TI - Correlation between serogroup and susceptibility to chloramphenicol, clindamycin, erythromycin, rifampicin and tetracycline among 308 isolates of Clostridium difficile. AB - The susceptibility to chloramphenicol, clindamycin, erythromycin, rifampicin and tetracycline of 308 isolates of Clostridium difficile from various origins was determined by a disc diffusion susceptibility testing and the results were compared with the serogroup of the strains. For the five antimicrobials, there was a clear-cut separation between susceptible and resistant strains. Some correlation between resistance and serogroup was found. Almost all of the 161 isolates of serogroups A, F, G, H and X were susceptible to all antibiotics. The 32 toxigenic isolates of serogroup C were characterized by a typical resistance pattern which could be used for typing purposes. Other serogroups showed variable patterns. The review of 64 cases of antibiotic associated diarrhoea showed that these differences in susceptibility could have clinical implications: all seven cases due to clindamycin were caused by a clindamycin resistant strain of serogroup C, whereas cases associated with other antibiotics were distributed among various serogroups. PMID- 3053560 TI - Haemophilus influenzae from four laboratories in one Canadian city. AB - Serotype, biotype and antimicrobial susceptibilities of 250 clinical isolates of Haemophilus influenzae from University, affiliated and community hospitals and a private laboratory were compared. For each drug, agar dilution susceptibility testing was compared to at least one other method (modified Kirby-Bauer and/or microdilution). Most isolates (86%) were non-typable, 10% were type b. Biotype II was most common (58%). The highest prevalence of serotype b (28%) was seen in the community hospital, which also had only 4% of all biotype III isolates. beta Lactamase production ranged from 20% (private laboratory) to 5% (affiliated hospital); it was higher among type b (23%), biotype II (17%), and from non respiratory (26%) than respiratory sites (8%). 51% of 35 beta-lactamase producers were found in the 24% of patients under age 6. Microdilution missed seven while agar dilution and disc diffusion detected all. All isolates were susceptible to cefamandole, cefuroxime, cotrimoxazole and chloramphenicol, 86%, 98%, 99% and 27% to ampicillin, cefaclor, tetracycline and erythromycin respectively. Microdilution is unreliable for detection of ampicillin resistance mediated by beta-lactamase production. PMID- 3053561 TI - A multi-infection model for antifungal screening in vivo. AB - A new in-vivo antifungal screen is described in which each mouse is given a vaginal infection with Candida albicans, a dermal infection with Trichophyton quinckeanum, a systemic infection with Can. albicans and a lung infection with Cryptococcus neoformans. Mice are dosed orally once daily on days 0-3 and infections evaluated on day 6 by visually scoring the dermal lesions and by culturing vaginal samples and kidney and lung homogenates. Mice carrying the multiple infections show no signs of distress at this time. Validation studies with eight antifungal agents show there is no interference between the four infections, and illustrate the different patterns of activity which result from differences in innate sensitivity and pharmacological behaviour of drugs in the host. This screen provides the maximum amount of information for a minimal investment of compound and effort, and naturally uses fewer animals than multiple tests using single infections. PMID- 3053562 TI - Amoxycillin-clavulanic acid (Augmentin) antibiotic prophylaxis against wound infections in renal failure patients. AB - A randomized, controlled trial of the use of amoxycillin with clavulanic acid (Augmentin) for prophylaxis against wound infections following major surgery, including transplantation, in patients with chronic renal failure, was undertaken. Six of 22 control patients developed wound infections (27%) whereas no patient in the treatment group (24) developed a wound infection (P less than 0.05). After the termination of this trial, the next 35 consecutive patients received prophylactic amoxycillin/clavulanate; of these only two developed wound infections associated with leakage from their pancreatic anastomoses. All the wound infections were shown to be caused by bacteria sensitive to amoxycillin/clavulanate. Pharmacokinetic studies in patients have shown that a bactericidal concentration of the drugs was present for up to 20 h post operatively in patients on dialysis, and in recipients of non-functioning renal transplants. In patients with normal renal transplant function excretion of the drug within 12 h was observed. PMID- 3053563 TI - Activity of metronidazole in mixed cultures. PMID- 3053564 TI - Early studies on in-vitro and experimental activity of spiramycin: a review. AB - This review of spiramycin activity in vitro is based mainly on early studies. The MICs of spiramycin for common pathogenic bacteria such as staphylococci, streptococci and pneumococci are higher than those of erythromycin. Conversely, in experimental models, the activity of spiramycin is equal to or greater than that of erythromycin. In addition, the activity of spiramycin on Neisseria, Legionella, Mycoplasma, Chlamydia, and Toxoplasma spp. completes its antimicrobial spectrum and shows that spiramycin covers the majority of agents responsible for respiratory tract infections. The 'spiramycin paradox'-the discrepancy between the relatively modest activity of spiramycin in vitro and its excellent activity in vivo will be explained by other papers. Its high tissue and intracellular concentrations, and the slow recovery of bacteria submitted to spiramycin are of great importance to account for its activity in vivo. PMID- 3053565 TI - Spiramycin concentrations in the human respiratory tract: a review. AB - Spiramycin has exceptionally good distribution properties, especially in respiratory tract tissues and fluids. Three hours after a single oral dose of 3 g spiramycin, the serum concentration ranged from 1.6 to 2.8 mg/l and the reported half-life was approximately 8 h. Studies of lung tissue concentrations showed that high pulmonary levels were achieved after a loading dose of 3 g; the levels were higher after multiple doses and reached approximately 30 to 45 mg/kg in lung tissue and 6.5 to 36 mg/kg in bronchial mucosa; in bronchial secretions and in sputum the concentrations of spiramycin ranged from 1.5 to 7.3 mg/l (after 1 g, multiple doses). In upper respiratory tract tissues and fluids, high levels of spiramycin were reached as well: 8 to 13 mg/kg in sinus mucosa; 15 to 29.5 mg/kg in tonsils or adenoids. PMID- 3053567 TI - The spiramycin paradox. AB - Spiramycin has been found to be effective in a variety of clinical and experimental infections despite modest in-vitro activity. In animal models of infection, spiramycin has been found to be as effective as or more effective than erythromycin despite inferior in-vitro activity. These paradoxical results are explained in part by spiramycin's ability to achieve intra-cellular and tissue concentrations that exceed serum concentrations by a factor of ten or more. Furthermore, spiramycin clearance from these sites is much lower resulting in sustained tissue and intracellular concentrations. Finally, spiramycin appears to produce a substantial post-antibiotic effect and, possibly, subinhibitory effects that may further enhance its in-vivo activity. PMID- 3053566 TI - Mechanism of action of spiramycin and other macrolides. AB - Macrolide antibiotics constitute a group of 12 to 16-membered lactone rings substituted with one or more sugar residues, some of which may be amino sugars. They inhibit bacterial protein synthesis both in vivo and in vitro with varying potencies. Macrolides are generally bacteriostatic, although some of these drugs may be bactericidal at very high concentrations. The mechanism of action of macrolides has been a matter of controversy for some time. Spiramycin, a 16 membered macrolide, inhibits translocation by binding to bacterial 50S ribosomal subunits with an apparent 1:1 stoichiometry. This antibiotic is a potent inhibitor of the binding to the ribosome of both donor and acceptor substrates. Spiramycin induces rapid breakdown of polyribosomes, an effect which has formerly been interpreted as occurring by normal ribosomal run-off followed by an antibiotic-induced block at or shortly after initiation of a new peptide. However, there is now convincing evidence that spiramycin, and probably all macrolides, act primarily by stimulating the dissociation of peptidyl-tRNA from ribosomes during translocation. Although the ribosomes of both Gram-positive and Gram-negative organisms are susceptible to macrolides, these antibiotics are mainly used against Gram-positive bacteria since they are unable to enter the porins of Gram-negative bacteria. Resistance to macrolides in clinical isolates is most frequently due to post-transcriptional methylation of an adenine residue of 23S ribosomal RNA, which leads to co-resistance to macrolides, lincosamides and streptogramins type B (the so-called MLSB phenotype). Other mechanisms of resistance involving cell impermeability or drug inactivation have been detected in Staphylococcus spp. and Escherichia coli. These strains are resistant to 14 membered macrolides (erythromycin and oleandomycin) but remain susceptible to spiramycin. PMID- 3053568 TI - The clinical use of macrolides. AB - Macrolides are active against Streptococcus pneumoniae, Legionella spp. and Mycoplasma pneumoniae, the main causes of community-acquired pneumonia They may therefore be used for the empirical treatment of community-acquired pneumonia, although emergent resistance in Str. pneumoniae limits their use in some parts of the world. In patients with bronchitis the use of macrolides reduces the severity and duration of symptoms. Macrolides have also been used successfully in the treatment of otitis media and sinusitis; combination with sulphonamides may be desirable. They may be effective in eradicating the carrier state of Str. pyogenes, Bordetella pertussis, Corynebacterium diptheriae, and Neisseria meningitidis. Macrolides provide alternative therapy for the prophylaxis of recurrent acute rheumatic fever and of infective endocarditis after dental treatment. The cure rate with macrolides of streptococcal skin infections and of minor staphylococcal infections is equal to that achieved with penicillins. In diarrhoea due to Campylobacter jejuni, the administration of macrolides shortens the duration of the faecal excretion of organisms and may give clinical improvement in severe disease. Macrolides are the drugs of choice for infections due to Chlamydia trachomatis in pregnancy and for Haemophilus ducreyi infections. They are effective alternative therapy to benzylpenicillin for the treatment of N. gonorrhoeae and Treponema pallidum infections. PMID- 3053569 TI - Comparison of spiramycin and doxycycline in the treatment of lower respiratory infections in general practice. AB - A total of 221 patients from 21 general practitioners was entered in a double blind comparative study of spiramycin and doxycycline in the treatment of pneumonia and acute exacerbations of chronic bronchitis. One-hundred-and-five patients were randomized to treatment with spiramycin tablets for 5 1/2 days and 116 patients were randomized to treatment with doxycycline tablets for nine days. The efficacy and side effects of the two treatment regimens were observed. Of the 221 patients included, 191 were acceptable for evaluation, 91 in the spiramycin group and 100 in the doxycycline group. Three patients in the spiramycin group withdrew because of lack of efficacy and one patient in the doxycycline group withdrew because of side effects (feeling unwell and blurred vision). No significant differences in efficacy or safety were found between the two treatments. PMID- 3053570 TI - Comparison of spiramycin with erythromycin for lower respiratory tract infections. AB - The efficacy and safety of 2 g oral spiramycin daily were compared with those of 2 g of oral erythromycin daily in a multicentre open prospective trial, involving 198 patients, with a mean age of 61.75 years, with a clinical and radiological diagnosis of acute lower respiratory tract infection. The diagnoses were: acute bronchitis (96), acute superinfection of chronic bronchitis (60) and pneumonia (42). The patients were assessed before therapy and after three and ten days of therapy. Seventy-four (76.3%) of the patients were cured in the spiramycin group and 64 (63.4%) were cured in the erythromycin group (P less than 0.05). Significantly more patients complained of side effects in the erythromycin group (41.4%) than in the spiramycin group (11.8%) P less than 0.001. PMID- 3053571 TI - The efficacy and safety of spiramycin in the treatment of nongonococcal urethritis in men. AB - Twenty-five male patients with nongonococcal urethritis including 15 chlamydial infections, were treated with spiramycin for ten days. All but four patients had been treated previously, mostly with tetracyclines. Chlamydia trachomatis was cultured in seven patients and was detected in three additional men by immunofluorescent smear. Five other patients had antibodies to chlamydia, and one patient yielded a positive culture for Ureaplasma urealyticum and Mycoplasma hominis. A successful clinical response was observed in 64% of the patients; C. trachomatis was eradicated from six of seven patients with positive cultures and the three positive direct smears were negative after treatment. It is concluded that spiramycin can be used effectively for the therapy of acute nongonococcal urethritis, as well as in patients who have failed to respond to previous treatment with tetracyclines and erythromycin. PMID- 3053572 TI - Macrolides and gastrointestinal motility. AB - Erythromycin was the first macrolide used clinically, and it is still the most widely prescribed in spite of reports of gastrointestinal side-effects. Erythromycin was given iv or orally to fasted and fed dogs with sensors implanted on the gastrointestinal tract for the measurement of motility. There was a large increase in stomach and upper small bowel contractile activity, accompanied by nausea and vomiting, while the distal small bowel appeared inhibited. Similar effects were seen in man. By contrast, two 16-membered macrolides, spiramycin and josamycin, did not produce such side-effects when given either orally or intravenously to dogs. PMID- 3053573 TI - Spiramycin: safety in man. AB - Spiramycin, a 16-membered lactone ring macrolide, has been in clinical use for the past 15 years with little serious associated toxicity. Gastrointestinal disturbance has usually been mild and no changes in gastrointestinal motility have been noted either experimentally or in humans, in contrast to other macrolides, such as erythromycin. Allergic reactions have been uncommon and mainly restricted to transient skin eruptions. Although liver injury is a possible complication of most macrolide treatments, no conclusive evidence for spiramycin-induced hepatitis is currently available, and, again in contrast to most other macrolides, the lack of drug interactions with spiramycin has been clearly established in biochemical, pharmacokinetic and clinical studies. PMID- 3053574 TI - Bactericidal mechanisms of ofloxacin. AB - Ciprofloxacin and ofloxacin are known to exert a second bactericidal mechanism (termed B) against Escherichia coli which functions even when protein synthesis is inhibited by chloramphenicol or when RNA synthesis is inhibited by rifampicin. However, the bactericidal activity of ciprofloxacin against a coagulase-negative staphylococcus (Staphylococcus warneri) was found to be abolished by chloramphenicol so the 4-quinolone does not exert mechanism B against this species. On the other hand, ofloxacin did exhibit mechanism B against S. warneri because the drug remained bactericidal in the presence of chloramphenicol. When S. aureus was investigated results similar to those observed in S. warneri were obtained throughout the range of clinically achievable concentrations of ofloxacin and ciprofloxacin. Ofloxacin seems to exhibit mechanism B against the staphylococci while ciprofloxacin does not. This may explain why ciprofloxacin is more potent than ofloxacin against Gram-negative bacteria but against staphylococci both drugs are equipotent. PMID- 3053575 TI - Interaction between the fluoroquinolones and the bronchodilator theophylline. AB - This review summarizes the available data on the influence of ofloxacin on the metabolic clearance of the bronchodilator theophylline. At the moment, several new fluoroquinolone derivatives, such as ofloxacin, ciprofloxacin, pefloxacin, and enoxacin are being clinically tested in respiratory tract infections. Enoxacin causes a strong and clinically important decrease (60%) of the total body clearance of theophylline. Ciprofloxacin and pefloxacin show the same effect, though to a smaller degree (30%). During treatment with these three agents clinical signs and symptoms of theophylline toxicity have been reported. However, no signs of increased plasma theophylline concentrations have been observed during concomitant treatment with ofloxacin and theophylline. Further research into the mechanism of this interaction has demonstrated that quinolones compete with cytochrome P450 related isoenzymes, resulting in a decreased demethylation of theophylline. Whereas a slight influence on these enzymes could be demonstrated for ofloxacin when the drug was administered in very high concentrations to rats, no significant influence on theophylline metabolic pathways in man has been measured, when ofloxacin was administered in doses up to 800 mg daily. PMID- 3053576 TI - Use of ofloxacin in open fractures and in the treatment of post-traumatic osteomyelitis. AB - In two open prospective studies, the efficacy and tolerance of ofloxacin in the prevention of infection in patients with open fractures (n = 58) and in the treatment of chronic post-traumatic osteomyelitis (n = 115) were examined. In the study with open fractures, bone and/or soft tissue infection occurred in only four cases (6.5%). During an observation period of at least 12 months, post traumatic osteomyelitis was seen in two patients with III degree open fractures (9%), while in the groups with I degree and II degree open fractures no bone infection could be found. Therefore, the rate of post-traumatic osteomyelitis related to all patients was 3.3%. In the second study with 115 patients suffering from chronic post-traumatic osteomyelitis 141 different Gram-positive and Gram negative pathogens were isolated. 73% were Gram-positive cocci with Staphylococcus aureus in more than 50% of the cases. An elimination rate of more than 90% was found in the Gram-positive and Gram-negative bacteria, leading to a clinical cure in 85% and a recurrence of infection in 5% of the cases. The tolerability of ofloxacin was excellent. No drug-related allergic reactions were observed. Diarrhoea and headache occurred in less than 2% of patients. With adequate surgical treatment, ofloxacin proved to be a useful antimicrobial agent in the prevention and therapy of bone infection. PMID- 3053577 TI - Overview of postmarketing experience with ofloxacin in Germany. AB - Clinical trials with ofloxacin have shown that adverse drug events (ADEs) occurred in between 2.4% (Phase II) and 3.1-7.3% (Phase IV) of patients treated and were mostly mild. As with any other drug the true spectrum of rare events can only be fully appreciated after marketing. Since the launch of ofloxacin in June 1985 about 3.5 million patients have been treated in Germany, calculated on the basis of a mean daily dose of 400 mg ofloxacin and a mean duration of treatment of seven days. During these 2.5 years 985 spontaneous national reports of ADEs have been obtained and include rare adverse events (e.g. hallucination, psychotic reaction and shock), not seen in clinical trials. The present status of results from postmarketing surveillance is shown and discussed. The favourable overall risk:benefit ratio of ofloxacin appears unchanged. PMID- 3053578 TI - A Belgian multicentre in-vitro study of ofloxacin. AB - In a large multicentre study, the susceptibility of 2171 bacterial isolates to ofloxacin and other antibiotics has been examined by the Kirby-Bauer method. Susceptibility to ofloxacin (breakpoint less than or equal to 2 mg/l) was observed in 99.2% of 1104 strains Enterobacteriaceae, 88.9% of 217 Pseudomonas aeruginosa, 93.6% of 31 other Pseudomonas spp., 96.3% of 27 Acinetobacter calcoaceticus, 99.2% of 118 Haemophilus influenzae, 97.4% of 423 staphylococci, 79.4% of 107 Streptococcus faecalis and 98.7% of 142 other streptococci. There was no relationship between resistance to ofloxacin and resistance to beta-lactam antibiotics, aminoglycosides or other antibiotics tested within the same bacterial species. PMID- 3053579 TI - Overview of drug interactions with the quinolones. AB - Drug interactions with the quinolones are of two types: pharmacokinetic and pharmacodynamic. Pharmacokinetic interactions include inhibition of absorption of quinolones by aluminium and magnesium containing antacids and inhibition of metabolism of other drugs by quinolones. Norfloxacin and ofloxacin are not extensively metabolized and do not inhibit drug metabolism; ciprofloxacin and enoxacin reduce theophylline clearance in normal subjects by less than 50% and greater than 50% respectively. Ciprofloxacin inhibits the metabolism of caffeine, theophylline and antipyrine. The latter is a marker of broad substrate specificity and, until proved otherwise, it would be prudent to avoid combination of ciprofloxacin with drugs which are metabolized and have a low therapeutic index. In addition to theophylline, these include cyclosporin, phenytoin and warfarin. There is evidence that the elderly and patients with liver disease are particularly susceptible to kinetic interactions with ciprofloxacin. In contrast, there is no evidence to suggest that ofloxacin is likely to impair hepatic drug elimination. Enoxacin does not impair the metabolism of chlorpropamide or glibenclamide, it is therefore unlikely that any of the quinolones will interact with sulphonylurea hypoglycaemic drugs. A pharmacodynamic interaction has been demonstrated in vitro between quinolones and non-steroidal anti-inflammatory drugs (NSAIDS) or theophylline. All of these drugs inhibit binding of radio labelled gamma-amino-butyric acid to mouse synaptic membranes and combinations of quinolones with NSAIDS or theophylline are synergistic. Convulsions have been reported in patients who received a combination of enoxacin with either fenbufen, a NSAID, or theophylline. Like theophylline, NSAIDS undergo hepatic metabolism, so the clinical interaction may be the result of combined pharmacokinetic and pharmacodynamic interactions. Drug-interactions with quinolones are a clinically important problem. Drugs, such as ofloxacin, which do not impair hepatic metabolism of other drugs, have a clinically significant advantage over other quinolones. The pharmacodynamic interaction between quinolones and other GABA inhibitors is extremely poorly documented; further in-vitro, animal and clinical studies are urgently required. PMID- 3053580 TI - Afferent neural pathways in cough and reflex bronchoconstriction. AB - Cough and bronchoconstriction are airway reflexes that protect the lung from inspired noxious agents. These two reflexes can be evoked both from the larynx and tracheobronchial tree and also from some extrarespiratory sites. Within the airways, certain sites are particularly sensitive to stimulation of cough (larynx and points of proximal airway branching), whereas bronchoconstriction can be triggered from the whole of the tracheobronchial tree. In the larynx, "irritant" receptors with myelinated afferents mediate cough and bronchoconstriction. Little seems to be known about laryngeal nonmyelinated afferents and their reflexes. In the tracheobronchial tree and lung, slowly adapting stretch receptors (SARs) and rapidly adapting stretch receptors (RARs) have opposing effects on airway tone, the former mediating bronchodilation and the latter bronchoconstriction. In cough, on the other hand, they operate concurrently, a mediatory role for RARs and a facilitatory role for SARs. C-fiber endings (bronchial and pulmonary) mediate bronchoconstriction. Inhalation of so-called "selective" C-fiber stimulants induces cough, but excitation of RARs has not been eliminated, and the possibility also exists that the cough is secondary to other lung actions mediated by these nerve endings. Although cough and bronchoconstriction may be mediated by the same type of receptor, they seem to have separate afferent neural pathways. PMID- 3053581 TI - Mechanism of reduced LV afterload by systolic and diastolic positive pleural pressure. AB - To investigate the mechanism by which increased pleural pressure (Ppl) assists left ventricular (LV) ejection, we compared the effects of phasic systolic or diastolic increases in Ppl (40-60 mmHg) with use of an isolated canine heart-lung preparation with constant venous return. Positive Ppl during systole (S) caused left atrial transmural pressure (Platm = Pla - Ppl) to decrease by 1.25 +/- 0.46 (SE) mmHg (P less than 0.025). Central blood volume (CBV), the volume of blood in the heart, lungs, and thoracic great vessels, decreased by 29 +/- 4.0 (SE) ml (P less than 0.001). When Ppl was raised for an equal duration during diastole (D), the decrease in Platm was not significant, but there was a significant decrease in CBV (10.5 +/- 4.1 ml, P less than 0.05). With constant venous return, these changes suggested that phasic elevations in Ppl in either S or D assisted LV ejection by decreasing LV afterload. To test the hypothesis that positive Ppl during D reduced afterload by emptying the thoracic aorta, we compared the effects of diastolic positive Ppl with a rigid aorta vs. a compliant aorta. Although there was no statistical difference in the effects of diastolic positive Ppl on Platm, the decrease in CBV was significantly greater when the aorta was compliant than when it was rigid (23 +/- 2.2 vs. 17 +/- 2.7 ml, P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3053582 TI - Effect of positive-pressure ventilatory frequency on regional pleural pressure. AB - Regional lung ventilation is modulated by the spatiotemporal distribution of alveolar distending forces. During positive-pressure ventilation, regional transmission of airway pressure (Paw) to the pleural surface may vary with ventilatory frequency (f), thus changing interregional airflow distribution. Pendelluft phenomena may result owing to selective regional hyperventilation or phase differences in alveolar distension. To define the effects of f on regional alveolar distension during positive-pressure ventilation, we compared regional pleural pressure (Ppl) swings from expiration to inspiration (delta Ppl) and end expiratory Ppl over the f range 0-150 min-1 in anesthetized, paralyzed, close chested dogs with normal lungs. We inserted six pleural balloon catheters to analyze Ppl distribution along three orthogonal axes of the right hemithorax. Increases in regional Ppl were synchronously coupled with inspiratory increases in Paw regardless of f. However, at a constant tidal volume and percent inspiratory time, end-expiratory Paw and Ppl increased in all regions once a f threshold was reached (P less than 0.01). Supradiaphragmatic delta Ppl were less than in other regions (P less than 0.05), but thoracoabdominal binding abolished this difference by decreasing thoracoabdominal compliance. We conclude that the distribution of forces determining dynamic regional alveolar distension are temporally synchronous but spatially asymmetric during positive-pressure ventilation at f less than or equal to 150/min. PMID- 3053583 TI - Impact of PEEP on lung mechanics and work of breathing in severe airflow obstruction. AB - Positive end-expiratory pressure (PEEP) has generally been withheld from the treatment of patients with chronic airflow obstruction (CAO), in view of the risk of hyperinflation and lack of documented benefit. We studied 10 mechanically ventilated patients with exacerbated CAO and air trapping to determine the impact of PEEP on lung mechanics, alveolar pressure, and the work of breathing. PEEP levels of 5 and 10 cmH2O were applied to patients whose end-expiratory alveolar pressures were documented to be positive when breathing against ambient pressure (the auto-PEEP effect). All patients were studied under two conditions: every breath machine assisted (AMV) and every breath machine controlled (paralyzed, CMV). PEEP improved expiratory resistance without substantially increasing peak static pressure. Inspiratory resistance remained unchanged. The difference between the end-expiratory values of alveolar and central airway pressure narrowed as PEEP increased. Adding PEEP improved the effective triggering sensitivity of the ventilator, diminished ventilatory drive, and reduced the mechanical work of breathing during the machine-assisted ventilatory cycle. Our results indicate that low levels of PEEP may improve lung mechanics and reduce the effort required of mechanically ventilated patients with severe airflow obstruction, without substantially increasing the hazards of hyperinflation. PMID- 3053584 TI - Pulmonary O2 toxicity in lambs: physiological and biochemical effects of endotoxin infusion. AB - Hyperoxic adult rats have prolonged survival and reduced morphological evidence of lung injury when treated with a single dose of bacterial endotoxin; this effect is mediated by an augmentation of antioxidant enzyme activity in lung homogenate. To determine whether endotoxin would prolong survival and influence antioxidant enzyme levels in lambs whose physiological response to O2 breathing can be serially measured, we administered a single intravenous dose of endotoxin (0.75 microgram/kg body wt) to 13 lambs before exposing them to greater than 95% O2 (n = 11) or air (n = 2). Seven additional lambs were placed in O2 after receiving only saline vehicle. All lambs had been instrumented to measure pulmonary vascular pressures and cardiac output, and 10 lambs had lung lymph fistulas. O2-exposed control lambs developed noncardiogenic pulmonary edema and respiratory failure within 85 +/- 10 h (range 76-110 h); antioxidant enzymes were not increased, but reduced glutathione (GSH) levels fell and oxidized glutathione (GSSG) increased, reflecting the oxidant stress of O2 exposure. By contrast, endotoxin-treated O2-exposed lambs had a delayed increase in microvascular permeability to protein, a reduced rate of lung edema formation, normal gas exchange after 72 h in O2, and prolonged survival (136 +/- 15 h; range 90-160 h; all variables P less than 0.05). Despite prolonged survival, postmortem lung water content was no greater in the lambs that received endotoxin. Treatment with endotoxin did not increase antioxidant enzyme levels in lung homogenate, but levels of GSH relative to GSSG were significantly elevated.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3053585 TI - Effect of endotoxin pretreatment on the pulmonary vascular response to hypoxia in O2-exposed lambs. AB - We recently reported that endotoxin infusion before O2 exposure significantly reduced or delayed the onset of pulmonary edema formation and respiratory failure by reducing the oxidant stress of O2 exposure. Despite these beneficial effects of endotoxin treatment, lung microvascular permeability eventually increased, but postmortem lung water content was less than expected. Prolonged O2 breathing blunts or abolishes the pulmonary constrictor response to alveolar hypoxia in some species, and it is possible that the loss of this response could contribute further to edema formation. To determine whether the reduction in lung edema observed in endotoxin-treated, O2-exposed lambs was linked to the preservation of hypoxic pulmonary vasoconstriction (HPV), we measured pulmonary vascular resistance before and after 8 min of isocarbic hypoxia (inspired O2 fraction 0.12) during each day of O2 exposure. In six control lambs, the pressor response to hypoxia was abolished after 72 h in O2, and the lambs developed respiratory failure shortly thereafter. In six endotoxin-treated lambs, HPV was preserved for as long as 144 h of O2 exposure. In two control O2-exposed lambs in whom HPV was abolished, the infusion of either angiotensin or prostaglandin H2 analogue increased pulmonary vascular resistance by greater than 75%. We conclude that in lambs 1) hyperoxia abolishes the pulmonary vascular response to hypoxia, 2) endotoxin pretreatment reduces acute O2-induced lung injury and preserves the pulmonary constrictor response to hypoxia, and 3) the loss of HPV during O2 exposure may be the result of oxidant-mediated injury to the hypoxia response itself and not the result of diffuse damage to the vasoconstrictor effector mechanism. PMID- 3053586 TI - Antigen challenge of sensitized rats increases airway responsiveness to methacholine. AB - We measured airway responsiveness to methacholine (MCh) of highly inbred rats before and after six inhalational challenges with antigen. Ten Brown-Norway rats (130-216 g) that were actively sensitized to ovalbumin (OA) received six challenges with OA at 5-day intervals beginning 19 days after sensitization. An aerosol of OA (5% wt/vol) was inhaled for 1, 2, 5, and 10 min or until pulmonary resistance (RL) increased by at least 50%. Challenges with aerosolized MCh were performed immediately before and 14 days after sensitization, 2 days after the 3rd OA exposure, and 2, 7, 12, and 17 days after the 6th OA challenge. Four unsensitized rats underwent inhalational challenges with MCh over an equivalent time period. Responsiveness to MCh was calculated as the concentration of MCh required to increase RL to 200% of the control value (EC200RL). Seven out of 10 rats in the experimental group reacted to the first OA challenge with an immediate increase in RL of greater than 50% of control (range 70-550%). Three animals were unreactive to OA. Base-line EC200RL for all rats undergoing sensitization was 2.13 mg/ml (geometric mean), and it did not change significantly after sensitization (2.05 mg/ml). However, EC200RL of the rats that reacted to OA (n = 7) decreased significantly after 3 (1.11 mg/ml; P less than 0.005) and 6 OA exposures (0.96 mg/ml; P less than 0.005). The increase in responsiveness to inhaled MCh was present 17 days after the last OA exposure (EC200RL = 1.40 mg/ml; P less than 0.05). EC200RL of neither the unreactive sensitized rats (n = 3) nor the control rats (n = 4) changed after OA challenges.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3053587 TI - Effect of carbohydrate feedings during high-intensity exercise. AB - To determine the upper limits of steady-state exercise performance and carbohydrate oxidation late in exercise, seven trained men were studied on two occasions during prolonged cycling that alternated every 15 min between approximately 60% and approximately 85% of VO2max. When fed a sweet placebo throughout exercise, plasma glucose and respiratory exchange ratio (R) declined (P less than 0.05) from 5.0 +/- 0.1 mM and 0.91 +/- 0.01 after 30 min (i.e., at 85% VO2max) to 3.7 +/- 0.3 mM and 0.79 +/- 0.01 at fatigue (i.e., when the subjects were unable to continue exercise at 60% VO2max). Carbohydrate feeding throughout exercise (1 g/kg at 10 min, then 0.6 g/kg every 30 min) increased plasma glucose to approximately 6 mM and partially prevented this decline in carbohydrate oxidation, allowing the men to perform 19% more work (2.74 +/- 0.13 vs. 2.29 +/- 0.09 MJ, P less than 0.05) before fatiguing. Even when fed carbohydrate, however, by the 3rd h of exercise, R had fallen from 0.92 to 0.87, accompanied by a reduction in exercise intensity from approximately 85% to approximately 75% VO2max (both P less than 0.05). These data indicate that carbohydrate feedings enable trained cyclists to exercise at up to 75% VO2max and to oxidize carbohydrate at up to 2 g/min during the later stages of prolonged intense exercise. PMID- 3053588 TI - Hindlimb suspension increases insulin binding and glucose metabolism. AB - After 28 days of hindlimb-suspension, insulin binding, 2-deoxy-D-glucose (2-DG) uptake, and glucose metabolism (glycolysis and glycogenesis) were determined at various insulin concentrations (0.2-30 nM) in soleus muscle of young (18-day-old) and adult (150-day-old) rats. Compared with age-matched controls the young (YS) and adult suspended (AS) rats had lower soleus and body weights and insulin levels (P less than 0.05). Per milligram of protein, insulin binding, 2-DG uptake, and the rate of glycolysis were increased by approximately 200%, and the rate of glycogenesis was increased approximately 100% in the YS group (P less than 0.05). Except for a reduction in glycogenesis (P less than 0.05) all other parameters also increased in the AS rats (P less than 0.05). On the basis of the whole muscle the rate of glucose metabolism (glycogenesis + glycolysis) was reduced in the YS rats (P less than 0.05), but in the AS rats glucose metabolism was similar to the controls. Thus the increased glucose metabolism (i.e., per milligram of protein) in the YS and AS groups may represent a compensatory response by atrophied muscle to attempt to sustain glucose removal from the circulation. Because greater insulin binding occurred in YS muscle [35% slow twitch (ST)] than in the control group (70% ST), and greater insulin binding occurred in the AS (81% ST) than in the control group (90% ST), higher insulin binding capacities are not always related to a high proportion of ST muscle fibers. In conclusion, after hindlimb suspension, marked increments in insulin binding and glucose metabolism occur in the soleus muscle. PMID- 3053590 TI - Treating malignancies with curative intent: understanding growth and differentiation. AB - Cancer is due to changes in the genes that normally regulate cellular growth and differentiation. These genetic changes cause cells to grow without normal regulation. Strategies for curative cancer treatment based on these new understandings are discussed. PMID- 3053589 TI - Effect of perivascular electromagnetic flow probes on pulmonary hemodynamics. AB - We determined the effect of perivascular electromagnetic flow probes (EMF) on pulmonary hemodynamics in acute experiments. In seven dogs placement of the EMF on the main pulmonary artery (MPA) increased pulmonary arterial pulse pressure by 25% (17.8-21.9 cmH2O, P less than 0.005) and mean right ventricular pressure by 12% (23.2-25.9 cmH2O, P less than 0.001) but did not alter heart rate, systemic blood pressure, mean pulmonary arterial pressure, or right ventricular end diastolic pressure. This response was not abolished by local application of lidocaine to the MPA. In three cats input impedance was calculated from measurements of pressure and flow in the MPA. Impedance was calculated with flow measured using an EMF and ultrasonic volume flow probe (USF), which avoids the constraining effect of the EMF. When flow was measured with an EMF rather than a USF, there was a significant difference in the impedance spectra (P less than 0.001), but it was only apparent in the moduli greater than six harmonics. We conclude that the EMF does affect right ventricular afterload in acute experiments and alters the measured input impedance. PMID- 3053591 TI - Intensive chemotherapy or chemoradiotherapy with autologous marrow support as treatment for patients with solid tumors. AB - The use of high-dose chemotherapy or chemoradiotherapy with autologous marrow support has had a major impact in the therapy of patients with hematologic malignancies and is now being studied as therapy for patients with solid tumors. In this article the current status of autologous marrow transplantation for solid tumors is reviewed, the major limitations of this approach are described, and several current areas of research aimed at overcoming these limitations are discussed. PMID- 3053592 TI - Results of whole abdominopelvic irradiation with nodal boost for patients with endometrial cancer at high risk of failure in the peritoneal cavity. A prospective clinical trial at the Mayo Clinic. AB - From August 1981 to December 1986, 47 patients with endometrial cancer, surgical stage I through IV, received adjuvant whole abdominopelvic irradiation with a nodal and vaginal boost. Median age was 66.5 years (range: 37 to 86 years). Twenty-two patients were stages I-II and 25 patients were stages III-IV. Thirty four patients (79 per cent) had positive peritoneal cytology, 29 patients (62 per cent) had deep myometrial involvement, 27 patients (58 per cent) had high-grade lesions, 18 patients (40 per cent) had either serous-papillary or adenosquamous histologic variants, and 10 patients (22 per cent) had residual disease of up to 2 cm remaining after operation, mostly in the form of nodal disease. Twenty-four patients (51 per cent) have had two or more laparotomies. Mean follow-up was 40.5 months (range: 17 to 81 months). The 5-year overall survival rate was 66 per cent, and the 5-year relapse-free survival rate was 77 per cent. Toxicity has been modest, usually of the acute type, and particularly evident in thin patients (weight below 115 pounds). PMID- 3053593 TI - Invasive bladder cancer. Strategies for cure and bladder preservation. AB - At the present time, standard therapy for invasive bladder cancer includes radical cystectomy and urinary diversion. Clearly, there is a population of patients who can be rendered tumor free by combinations of irradiation and chemotherapy, or irradiation and surgery. These patients may not require total cystectomy to be cured. Only time and experience will tell which treatment regimens can provide both cure and preservation of the urinary bladder. PMID- 3053594 TI - Chemotherapy in ovarian carcinoma: intravenous or intraperitoneal? AB - Although the majority of women with advanced ovarian carcinoma respond to cisplatin-based intravenous chemotherapy, long-term survival is experienced by less than 30 per cent of treated patients. Efforts to improve the efficacy of treatment have focused on dose intensification and direct drug delivery into the peritoneal cavity. PMID- 3053595 TI - Testicular germ-cell neoplasms: curative approaches. AB - Approximately 80 per cent of all patients with testis cancer will present with low-volume disease. If they are pathologic stage I, there is virtually a 100 per cent cure rate even though 5 to 10 per cent will relapse after node dissection. Those who have relapsed have been salvaged with systemic chemotherapy. If clinical stage I patients are followed on observation protocols, 20 to 30 per cent of the patients will relapse. Unfortunately, a small percentage of these patients will not be salvaged with chemotherapy because they have more advanced disease at the time that the relapse is discovered. The main objection to retroperitoneal lymph node dissection in patients with low-stage disease has been the issue of infertility induced by the procedure. This problem has been alleviated by the development of the prospective nerve-sparing procedures. Long term follow-up is still required to make certain that the relapse rate is not significantly higher than with a full retroperitoneal lymph node dissection. Experience has demonstrated that patients with stage II disease will achieve a 98 to 99 per cent cure rate regardless of whether they have adjuvant chemotherapy after surgery, or full-course chemotherapy should they relapse (approximately 50 per cent). Advanced disease is best treated initially by systemic chemotherapy. The emphasis in the last few years has been on a reduction in the toxicity of chemotherapy while maintaining the high degree of efficacy. Surgery is used as an adjunct to chemotherapy in those patients who achieve a partial remission. Careful follow-up cannot be over emphasized in patients with any form of treatment for testis cancer. It is especially important that those patients who have teratoma present in the surgical specimens obtained at the time of a postchemotherapy lymph node dissection have prolonged follow-up, because recurrences have been noted as long as 6 to 8 years after the initial event. Although primary retroperitoneal or mediastinal germ-cell tumors do exist, the vast majority arise from the testis. If the diagnosis of germ-cell tumor has been made by some means other than orchiectomy, a careful review of the patient's history and physical findings is needed to guide the physician to the possible testicular site of origin. If no physical findings are present, scrotal ultrasound may be helpful in localizing a primary tumor of the testis. If such localization is possible, then a radical orchiectomy should be performed to prevent future seeding from persistent tumor in the testicle.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3053596 TI - Serum markers in testicular germ-cell neoplasms. AB - At the present time, serum determinations of alpha fetoprotein and beta-HCG are the most useful markers in the management of germ-cell testis tumors. Serial determinations are vital to the monitoring of the response of the patient's tumor to therapy. Subsequent elevation of tumor markers is often the first evidence of recurrence of tumor and allows effective treatment to be instigated while the volume of recurrent tumor is extremely low. This markedly increases the likelihood of achieving complete remissions. PMID- 3053597 TI - A simple medium for the study of hepatocyte growth in culture under defined conditions. AB - The combination (1:1) of Dulbecco's modified Eagle's medium and Waymouth's medium MAB 87/3 was found to provide favorable conditions for serum-free culture and growth of adult rat hepatocytes. In this simple medium, a majority of hepatocytes stimulated by epidermal growth factor plus insulin entered S phase and divided, with a normal (13 h) interval between DNA synthesis and cell division. The proliferative response did not require extra substratum or the presence of serum, even during cell isolation and plating. PMID- 3053600 TI - Identification of subgroups of childhood asthmatics: a review. AB - Given the literature suggesting etiological variability among asthmatics, the research pertaining to the identification of etiological subgroups was reviewed. The most promising prospects are: 1) the presence or absence of emotional precipitants of asthmatic symptoms and 2) the emotional reaction of the patient to his symptoms (e.g., "panic-fear"). These two dimensions have been found to be uncorrelated. The two studies that explored the treatment implications of these factors produced mixed results. Nonetheless, it may be promising to pursue the notion of identifiable subgroups that respond differently to psychological intervention. PMID- 3053599 TI - An interview with Albert Levin [by Barbara Katzenberg]. PMID- 3053598 TI - Intracellular protein degradation in cultured bovine lens epithelial cells. AB - Although several proteases have been identified in homogenates of cultured epithelial cells of the eye lens and in lens tissues, there is little information regarding intracellular protein degradation in intact lens cells in vitro. Cultured lens cells may be useful in the study of intracellular protein degradation in the lens, a tissue with a wide range of protein half-lives. This is of interest because alterations in protein turnover in the lens have been implicated in cataract formation. This study examines intracellular protein degradation in cultured bovine lens epithelial cells (BLEC). Cell cultures were incubated with radiolabeled leucine to label intracellular proteins. Protein degradation was measured by monitoring the release of trichloroacetic-acid soluble radioactivity into the culture medium. The average half-life of long lived proteins (half-life greater than 50 h) was typically about 57 h in serum supplemented medium. Average rates of degradation of long-lived proteins increased by up to 73% when fetal bovine serum was withdrawn from the culture medium. Serum had no effect on the degradation of short-lived proteins (half-life less than 10 h). Degradation of long-lived proteins in the presence and absence of serum was further studied in cultured BLEC from population doubling level (PDL) 2 to 43. Average half-life of proteins in serum-supplemented medium was 52 to 58 h and did not vary significantly as a function of PDL. Degradation rates in serum-free medium increased approximately twofold up to PDL 7, but returned by PDL 25 to original levels, which were maintained through PDL 43. PMID- 3053601 TI - Steady-state evaluation of two controlled-release theophylline preparations in children. AB - The absorption profiles of Theolair-SR and Theo-Dur were studied in 23 asthmatic children (mean age 11 years). A 250-mg dose of each product was administered twice a day until steady state was attained (mean, 8 days). Both products maintained adequate theophylline levels over 12 hours, although Theolair-SR gave significantly higher serum levels and a 19% greater extent of theophylline absorption. The mean Cmax-Cmin was also significantly greater for Theolair-SR. Both products should be considered comparable for practical use. PMID- 3053603 TI - Asthma self-management programs: premises, not promises. PMID- 3053602 TI - The clinical evaluation of nonbronchodilator antiasthmatic drugs. PMID- 3053604 TI - Seneca and his asthma: the illnesses, life, and death of a Roman stoic philosopher. PMID- 3053605 TI - High-dose methylprednisolone as initial therapy in patients with acute bronchospasm. AB - The use of steroids in treating acute respiratory obstruction is still controversial. In this double-blind controlled trial, we decided to examine the beneficial effects of a single large dose of methylprednisolone (MSSP), using objective criteria. In the emergency setting, methylprednisolone (30 mg/kg) has been shown to decrease the need for hospital admission in patients with acute bronchospasm. No difference in this improvement was seen among patients in the steroid-dependent or non-steroid-dependent populations. Based on our findings, we suggest that the early use of single-dose steroid therapy is appropriate treatment for patients with acute bronchospastic attacks. PMID- 3053606 TI - The theophylline-erythromycin interaction. AB - Since its publication in 1976, the original report of an interaction between erythromycin and theophylline by Cummin, Kozak, and Gillman has generated considerable interest and controversy. Many studies with considerably different designs have been performed to address this question. Those studies that most closely simulate the clinical setting suggest that a 7- to 10-day course of concurrent theophylline and erythromycin therapy will result in variable changes in theophylline clearance. It may be that as many as 25% of patients, especially when maintained with serum theophylline concentrations at the upper portion of the therapeutic range, display elevations in serum theophylline concentrations that might lead to clinical symptoms of theophylline toxicity. There has been a suggestion, based on the mean changes in several studies, that the interaction may lead to a 25% increase in serum theophylline concentrations; however, it is clear that there may be a much larger increase in some patients. This toxicity can be anticipated and avoided if careful attention is paid to monitoring the serum theophylline concentrations of such high-risk patients when erythromycin therapy is contemplated as an addition to theophylline therapy. Other macrolide antibiotics may display interactions with theophylline, which may be due in part to the ability of the various antibiotics to form complexes with isoenzymes of the cytochromes P-450. The growing impression of the importance of mycoplasma in asthmatics and the introduction of new macrolides onto the market make the appreciation of this possible interaction of extreme importance to primary care and chest physicians. PMID- 3053607 TI - A study of buspirone coprescribed with bronchodilators in 82 anxious ambulatory patients. AB - This was an open multicenter study wherein patients requiring anxiolytic therapy were treated with buspirone 5 mg t.i.d. for 4 weeks. A subgroup of 82 patients was concomitantly treated with bronchodilators. Theophylline and terbutaline were the most frequently used preparations. A matching cohort of 82 patients who used no concomitant bronchodilators was retrospectively selected as the control. Both groups of patients showed substantial and comparable improvement as measured by the physician and patient ratings. The bronchodilator group had an incidence of adverse effects (16%) slightly higher than that of the control group (12%), due mainly to a higher incidence of dizziness (8.5% versus 2.4%). The results indicate that buspirone is effective and well tolerated when taken in combination with a wide variety of bronchodilator medications. PMID- 3053608 TI - Changes in inpatient child psychiatry: consequences and recommendations. PMID- 3053609 TI - Efficacy and safety of fenfluramine in autistic children. PMID- 3053610 TI - Minor physical anomalies as a biologic marker for behavior disorders. PMID- 3053611 TI - Issues in the classification of child and adolescent psychopathology. PMID- 3053612 TI - Post-traumatic stress disorder in young children: a reaction to purported sexual abuse. PMID- 3053613 TI - Campylobacter pyloridis is not responsible for duodenal ulcer formation: results of a controlled therapeutic trial. PMID- 3053615 TI - Post-transplant diabetes in Indian renal allograft recipients. PMID- 3053614 TI - Antiautonomic nerve antibodies in autonomic neuropathy of diabetes mellitus. PMID- 3053616 TI - Autonomic innervation of human airways: its clinical significance. PMID- 3053617 TI - Role of complement in viral infections. PMID- 3053619 TI - Newer insulins. PMID- 3053618 TI - Corticosteroids in asthma: pharmacology and therapeutics. PMID- 3053620 TI - Effects of sucralfate on gastric acid secretion and duodenal ulcer healing. PMID- 3053621 TI - Comparison of ranitidine 300 mg nocte and 150 mg twice daily in the treatment of duodenal ulcer. PMID- 3053622 TI - Occurrence of urticaria as a manifestation of Falciparum malaria. PMID- 3053623 TI - Liver transplantation: medical aspects. PMID- 3053624 TI - Hypertension the other side of the coin. PMID- 3053625 TI - Sternal involvement in disseminated tuberculosis. PMID- 3053626 TI - Malarial hepatitis. PMID- 3053627 TI - Falciparum malaria presenting as testicular pain and swelling--a rejoinder. PMID- 3053628 TI - Atenolol vs captopril as monotherapy in mild to moderate hypertension: alternative to empirical step care approach. PMID- 3053629 TI - Doppler evaluation in carotid disease. PMID- 3053630 TI - Extraintestinal manifestations of Shigellosis during an epidemic of bacillary dysentery in Port Blair, Andaman & Nicobar Island (India). PMID- 3053631 TI - Low sensitisation rate with donor specific transfusion. PMID- 3053632 TI - Ultrasound guided kidney biopsy. PMID- 3053633 TI - Current status of autoimmunity in endocrinopathies. PMID- 3053634 TI - Nifedipine and pulmonary functions in bronchial asthma. PMID- 3053635 TI - Prolonged survival in chronic granulocytic leukaemia. PMID- 3053636 TI - Periodic peritonitis (recurrent polyserositis-or-familial Mediterranean fever) eight decades later. PMID- 3053637 TI - Diabetic nephropathy. PMID- 3053638 TI - Shigella flexneri bacteraemia in post partum period. PMID- 3053639 TI - What is the bacterial growth law during the division cycle? PMID- 3053640 TI - Induction of the autolytic system of Escherichia coli by specific insertion of bacteriophage MS2 lysis protein into the bacterial cell envelope. AB - Bacterial lysis induced by the expression of the cloned lysis gene of the RNA bacteriophage MS2 in Escherichia coli was shown to be under the same regulatory control mechanisms as penicillin-induced lysis. It was controlled by the stringent response and showed the phenomenon of tolerance when E. coli was grown at pH 5. Changes in the fine structure of the murein were found to be the earliest physiological changes in the cell, taking place 10 min before the onset of cellular lysis and inhibition of murein synthesis. Both the average length of the glycan strands and, with a time lag, the degree of cross-linkage were altered, indicating that a lytic transglycosylase and a DD-endopeptidase had been triggered. After extensive separation of the membranes by isopycnic sucrose gradient centrifugation, the lysis protein was present predominantly in the cytoplasmic membrane and in a fraction of intermediate density and, to a lesser degree, in the outer membrane, irrespective of the conditions of growth. However, only under lysis-permissive conditions could a 17% increase in the number of adhesion sites between the inner and outer membranes be observed. Thus, a casual relationship between lysis and the formation of lysis protein-induced adhesion sites seems to exist. PMID- 3053641 TI - Effects of induction of rRNA overproduction on ribosomal protein synthesis and ribosome subunit assembly in Escherichia coli. AB - Overproduction of rRNA was artificially induced in Escherichia coli cells to test whether the synthesis of ribosomal protein (r-protein) is normally repressed by feedback regulation. When rRNA was overproduced more than twofold from a hybrid plasmid carrying the rrnB operon fused to the lambda pL promoter (pL-rrnB), synthesis of individual r-proteins increased by an average of about 60%. This demonstrates that the synthesis of r-proteins is repressed under normal conditions. The increase of r-protein production, however, for unknown reasons, was not as great as the increase in rRNA synthesis and resulted in an imbalance between the amounts of rRNA and r-protein synthesis. Therefore, only a small (less than 20%) increase in the synthesis of complete 30S and 50S ribosome subunits was detected, and a considerable fraction of the excess rRNA was degraded. Lack of complete cooperativity in the assembly of ribosome subunits in vivo is discussed as a possible explanation for the absence of a large stimulation of ribosome synthesis observed under these conditions. In addition to the induction of intact rRNA overproduction from the pL-rrnB operon, the effects of unbalanced overproduction of each of the two large rRNAs, 16S rRNA and 23S rRNA, on r-protein synthesis were examined using pL-rrnB derivatives carrying a large deletion in either the 23S rRNA gene or the 16S rRNA gene. Operon-specific derepression after 23S or 16S rRNA overproduction correlated with the overproduction of rRNA containing the target site for the operon-specific repressor r-protein. These results are discussed to explain the apparent coupling of the assembly of one ribosomal subunit with that of the other which was observed in earlier studies on conditionally lethal mutants with defects in ribosome assembly. PMID- 3053643 TI - Evidence in vivo for autogenous control of the cyclic AMP receptor protein gene (crp) in Escherichia coli by divergent RNA. AB - Control of crp expression in vivo was studied by using the cloned crp gene. The synthesis of the product of this gene, cyclic AMP (cAMP) receptor protein (CRP), was strongly reduced by exogenous cAMP. This regulation was completely abolished by the inactivation of a divergent promoter located within the crp promoter region. These data are consistent with our in vitro studies (Okamoto and Freundlich, Proc.Natl. Acad. Sci. USA 83:5000-5004, 1986), which showed that crp autoregulation is due to the inhibition of crp transcription by divergent (antisense) RNA produced by cAMP-CRP activation of the divergent promoter. PMID- 3053642 TI - Degradation of a signal peptide by protease IV and oligopeptidase A. AB - The degradation of the prolipoprotein signal peptide in vitro by membranes, cytoplasmic fraction, and two purified major signal peptide peptidases from Escherichia coli was followed by reverse-phase liquid chromatography (RPLC). The cytoplasmic fraction hydrolyzed the signal peptide completely into amino acids. In contrast, many peptide fragments accumulated as final products during the cleavage by a membrane fraction. Most of the peptides were similar to the peptides formed during the cleavage of the signal peptide by the purified membrane-bound signal peptide peptidase, protease IV. Peptide fragments generated during the cleavage of the signal peptide by protease IV and a cytoplasmic enzyme, oligopeptidase A, were identified from their amino acid compositions, their retention times during RPLC, and knowledge of the amino acid sequence of the signal peptide. Both enzymes were endopeptidases, as neither dipeptides nor free amino acids were formed during the cleavage reactions. Protease IV cleaved the signal peptide predominantly in the hydrophobic segment (residues 7 to 14). Protease IV required substrates with hydrophobic amino acids at the primary and the adjacent substrate-binding sites, with a minimum of three amino acids on either side of the scissile bond. Oligopeptidase A cleaved peptides (minimally five residues) that had either alanine or glycine at the P'1 (primary binding site) or at the P1 (preceding P'1) site of the substrate. These results support the hypothesis that protease IV is the major signal peptide peptidase in membranes that initiates the degradation of the signal peptide by making endoproteolytic cuts; oligopeptidase A and other cytoplasmic enzymes further degrade the partially degraded portions of the signal peptide that may be diffused or transported back into the cytoplasm from the membranes. PMID- 3053644 TI - Purification and characterization of the wild-type and mutant carboxy-terminal domains of the Escherichia coli Tar chemoreceptor. AB - The carboxy-terminal half of the Escherichia coli Tar chemoreceptor protein was cloned into an overproducing plasmid with the transcription of the insert under the control of the strong hybrid tac promoter. Two dominant mutations in the tar gene, which result in "tumble-only" (tar-526) or "swim-only" (tar-529) phenotypes and which are postulated to produce proteins locked in specific signalling modes, were introduced separately onto the overproducing plasmid. After induction with isopropyl-beta-D-thiogalactopyranoside, cells containing the plasmids produced about 10% of their soluble cellular protein as the carboxy-terminal fragments. A scheme to purify the overproduced fragments was developed. Typical yields of pure fragment were 5, 30, and 20 mg per liter of induced culture for the wild type, 526 mutant, and 529 mutant, respectively. Fast-protein liquid chromatography-gel filtration analysis of the pure fragments showed that they all existed as oligomers (ca. 103,000 daltons), possibly trimers or tetramers (monomer size is 31,000 daltons). However, the 529 mutant fragment showed an additional oligomeric form (240,000 daltons) corresponding approximately to an octamer. When chromatographed in the presence of 1% octylglucoside, all three fragments showed an identical single oligomeric size of about 135,000 daltons. Further differences between the fragments such as ion-exchange behavior and susceptibility to degradation were found. Taken together, these results suggest that conformational differences between the 529 mutant fragment and the other fragments exist and that these differences may correlate with the phenotypic effects of the tar-529 mutation. PMID- 3053645 TI - Influence of growth temperature and lipopolysaccharide on hemolytic activity of Serratia marcescens. AB - Log-phase cells of Serratia marcescens cultured at 30 degrees C were approximately 10-fold more hemolytic than those grown at 37 degrees C. By using a cloned gene fusion of the promoter-proximal part of the hemolysin gene (shlA) to the Escherichia coli alkaline phosphatase gene (phoA), hemolysin gene expression as a function of alkaline phosphatase activity was measured at 30 and 37 degrees C. No difference in alkaline phosphatase activity was observed as a function of growth temperature, although more hemolysin was detectable immunologically in whole-cell extracts of cells grown at 30 degrees C. The influence of temperature was, however, growth phase dependent, because the hemolytic activities of cells cultured to early log phase at 30 and 37 degrees C were comparable. Given the outer membrane location of the hemolysin, lipopolysaccharide (LPS) was examined as a candidate for mediating the temperature effect on hemolytic activity. Silver staining of LPS in polyacrylamide gels revealed a shift towards shorter O-antigen molecules at 37 degrees C relative to 30 degrees C. Moreover, there was less binding of O-antigen-specific bacteriophage to S. marcescens with increasing growth temperature, a finding consistent with temperature-mediated changes in LPS structure. Smooth strains of S. marcescens were 20- to 30-fold more hemolytic than rough derivatives, a result confirming that changes in LPS structure can influence hemolytic activity. The alkaline phosphatase activity of rough strains harboring the shlA-phoA fusion was threefold lower than that of smooth strains harboring the fusion plasmids, a result consistent with a decrease in hemolysin gene expression in rough strains. The absence of a similar effect of temperature on gene expression may be related to less-marked changes in LPS structure as a function of temperature compared with a smooth-to-rough mutational change. PMID- 3053646 TI - Divergence of the aerobactin iron uptake systems encoded by plasmids pColV-K30 in Escherichia coli K-12 and pSMN1 in Aerobacter aerogenes 62-1. AB - Although the aerobactin-mediated iron uptake system has been characterized genetically in Escherichia coli, the siderophore aerobactin was chemically characterized after purification from culture supernatants of Aerobacter aerogenes 62-1, a member of the Klebsielleae. We have cloned and mapped the genes encoding the aerobactin system genes of A. aerogenes 62-1 and begun characterization of the relevant proteins and enzymatic activities of this plasmid-mediated aerobactin system. Published chemical data indicate that the siderophore aerobactin of E. coli is the same molecule as the aerobactin of Aerobacter aerogenes 62-1, but we have found that both the genes and the complement of proteins making up the biosynthetic enzymes in the two systems have diverged. In contrast, the outer membrane receptors for ferric aerobactin of the two systems showed immunologic cross-reactivity, were of the same molecular size (74 kilodaltons), and were encoded by homologous DNA sequences. PMID- 3053647 TI - Identification, cloning, and expression of bolA, an ftsZ-dependent morphogene of Escherichia coli. AB - A newly found morphogene of Escherichia coli, bolA, mapping at min 10 of the genetic map, was cloned in a 7.2-kilobase BamHI fragment and identified by its ability to produce osmotically stable spherical cells when overexpressed. This gene codes for a polypeptide of 13 kilodaltons. Overexpression of bolA+ was achieved in low-copy-number vectors with operon fusions to the tet and lac promoters, indicating a clockwise direction of transcription. While no modification of any of the penicillin-binding proteins was observed, morphological effects due to overexpression of bolA+ were shown to be dependent on the presence of an active ftsZ gene product. Our results suggest the existence of a mechanism mediated by FtsZ for modifying the conformation of nascent murein in the early steps of septum formation. PMID- 3053648 TI - Molecular cloning, sequencing, and mapping of the bacteriophage T2 dam gene. AB - Bacteriophage T2 codes for a DNA-(adenine-N6)methyltransferase (Dam), which is able to methylate both cytosine- and hydroxymethylcytosine-containing DNAs to a greater extent than the corresponding methyltransferase encoded by bacteriophage T4. We have cloned and sequenced the T2 dam gene and compared it with the T4 dam gene. In the Dam coding region, there are 22 nucleotide differences, 4 of which result in three coding differences (2 are in the same codon). Two of the amino acid alterations are located in a region of homology that is shared by T2 and T4 Dam, Escherichia coli Dam, and the modification enzyme of Streptococcus pneumoniae, all of which methylate the sequence 5' GATC 3'. The T2 dam and T4 dam promoters are not identical and appear to have slightly different efficiencies; when fused to the E. coli lacZ gene, the T4 promoter produces about twofold more beta-galactosidase activity than does the T2 promoter. In our first attempt to isolate T2 dam, a truncated gene was cloned on a 1.67-kilobase XbaI fragment. This construct produces a chimeric protein composed of the first 163 amino acids of T2 Dam followed by 83 amino acids coded by the pUC18 vector. Surprisingly, the chimera has Dam activity, but only on cytosine-containing DNA. Genetic and physical analyses place the T2 dam gene at the same respective map location as the T4 dam gene. However, relative to T4, T2 contains an insertion of 536 base pairs 5' to the dam gene. Southern blot hybridization and computer analysis failed to reveal any homology between this insert and either T4 or E. coli DNA. PMID- 3053650 TI - Enhanced acetohydroxy acid synthase III activity in an ilvH mutant of Escherichia coli K-12. AB - The acetohydroxy acid synthase III isozyme, which catalyzes the first common step in the biosynthesis of isoleucine, leucine, and valine in Escherichia coli K-12, is composed of two subunits, the ilvI and ilvH gene products. A missense mutation in ilvH (ilvH612), which reduced the sensitivity of the enzyme to the end product inhibition by valine, also increased its specific activity and lowered the Km for alpha-acetolactate synthesis. The mutation increased the sensitivity of acetohydroxy acid synthase III to dialysis and heat treatment and reduced the requirement for thiamine pyrophosphate addition to the assay mixture for activity. A strain carrying the ilvH612 mutation grew better than a homologous ilvH+ strain in the presence of leucine. The data indicate that this is a consequence of a more active acetohydroxy acid synthase III isozyme rather than the result of an alteration of the leucine-mediated repression of the ilvIH operon. PMID- 3053649 TI - Proline carrier mutant of Escherichia coli K-12 with altered cation sensitivity of substrate-binding activity: cloning, biochemical characterization, and identification of the mutation. AB - Two putP mutants of Escherichia coli K-12 that were defective in proline transport but retained the binding activities of the major proline carrier were isolated (T. Mogi, H. Yamamoto, T. Nakao, I. Yamato, and Y. Anraku, Mol. Gen. Genet. 202:35-41, 1986). One of these mutations and three null-type mutations (K. Motojima, I. Yamato, and Y. Anraku, J. Bacteriol. 136:5-9, 1978) were cloned into a pBR322 putP+ hybrid plasmid (pTMP5) by in vivo recombination. Cytoplasmic membrane vesicles were prepared from the mutant strains and strains harboring pTMP5 putP plasmids, and the properties of the proline-binding reaction of the mutant putP carriers in membranes were examined under nonenergized conditions. The putP19, putP21, and putP22 mutations, which were mapped in the same DNA segment of the putP gene (Mogi et al., Mol. Gen. Genet. 202:35-41, 1986), caused the complete loss of proline carrier activity. The proline carriers encoded by the mutant putP genes, putP9 and putP32, and putP32 in pTMP5-32, which was derived from in vivo recombination with the putP32 mutation, had altered sodium ion and proton dependence of binding affinities for proline and were resistant to N-ethylmaleimide inactivation without changes in the specificities for substrates and alkaline metal cations. The nucleotide sequence of the putP32 lesion located on the 0.35-megadalton RsaI-PvuII fragment in the putP gene in pTMP5-32 was determined; the mutation changed a cytosine at position 1001 to a thymine, causing the alteration of arginine to cysteine at amino acid position 257 in the primary structure of the proline carrier. It was shown that this one point mutation was enough to produce the phenotype of pTMP5-32 by in vitro DNA replacement of the AcyI-PvuII fragment of the wild-type putP gene with the DNA fragment containing the mutated nucleotide sequence. PMID- 3053651 TI - Utilization by Escherichia coli of a high-molecular-weight, linear polyphosphate: roles of phosphatases and pore proteins. AB - We observed that wild-type Escherichia coli utilized a linear polyphosphate with a chain length of 100 phosphate residues (poly-P100) as the sole source of phosphate in growth medium. A mutation in the gene phoA of alkaline phosphatase or phoB, the positive regulatory gene, prevented growth in this medium. Since no alkaline phosphatase activity was detected outside the wild-type cells, the periplasmic presence of the enzyme was necessary for the degradation of polyphosphate. A 90% reduction in the activity of periplasmic acid phosphatase with a pH optimum of 2.5 (delta appA mutants) did not affect polyphosphate utilization. Of the porins analyzed (OmpC, OmpF, and PhoE), the phoB-inducible porin PhoE was not essential since its absence did not prevent growth. To study how poly-P100 diffused into the cells, we used high-resolution 31P nuclear magnetic resonance (31P NMR) spectroscopy. The results suggest that poly-P100 entered the periplasm and remained in equilibrium between the periplasm and the medium. When present individually, porins PhoE and OmpF facilitated a higher permeability for poly-P100 than porin OmpC did. The degradation of polyphosphate by intact cells of E. coli observed by 31P NMR showed a time-dependent increase in cellular phosphate and a decrease in polyphosphate concentration. PMID- 3053652 TI - Thermosensitive omsA mutation of Escherichia coli that causes thermoregulated release of periplasmic proteins. AB - A mutant of Escherichia coli with a thermosensitive defect, possibly in the outer membrane (omsA mutant), was isolated from E. coli K-12 by mutagenization and selection for thermosensitivity and beta-lactam supersensitivity of growth. The mutant also showed very high sensitivity to other antibiotics, such as macarbomycin, midecamycin, rifampin, and bacitracin. The mutation was recessive to the wild type and was mapped at about 4 min on the E. coli chromosome between fhuA and metD. The mutation caused rapid release into the medium of periplasmic enzymes such as RTEM penicillinase but practically no cytoplasmic enzyme when cells grown at 30 degrees C were transferred to 37 or 42 degrees C. Electron microscopic observations showed many large double-layered vesicles attached to the surface of cells incubated at 42 degrees C. We conclude that the mutant had a mutation that caused a temperature-dependent defect in the outer membrane structure or its assembly (named an oms mutation). The omsA mutant may be useful for production of periplasmic proteins, which it releases into the culture medium on shift up of temperature. PMID- 3053653 TI - Mutagenic nucleoside analog N4-aminocytidine: metabolism, incorporation into DNA, and mutagenesis in Escherichia coli. AB - N4-Aminocytidine, a nucleoside analog, is strongly mutagenic to various organisms including Escherichia coli. Using E. coli WP2 (trp), we measured the incorporation of [5-3H]N4-aminocytidine into DNA and at the same time measured the frequency of reversion of the wild type, thereby attempting to correlate the incorporation with mutation induction. First, we observed that N4-aminocytidine uptake by the E. coli cells was as efficient as cytidine uptake. High-pressure liquid chromatographic analysis of nucleoside mixtures obtained by enzymatic digestion of isolated cellular DNA showed that the DNA contained [3H]N4 aminodeoxycytidine, corresponding to 0.01 to 0.07% of the total nucleoside; the content was dependent on the dose of N4-aminocytidine. There was a linear relationship between the N4-aminocytosine content in DNA and the mutation frequency observed. These results constitute strong evidence for the view that the N4-aminocytidine-induced mutation in E. coli is caused by the incorporation of this agent into DNA as N4-aminodeoxycytidine. We also found that the major portion of radioactivity in DNA of cells that had been treated with [5-3H]N4 aminocytidine was in the deoxycytidine fraction. We propose a metabolic pathway for N4-aminocytidine in cells of E. coli. This pathway involves the formation of both N4-aminodeoxycytidine 5'-triphosphate and deoxycytidine 5'-triphosphate; the deoxycytidine 5'-triphosphate formation is initiated by conversion of N4 aminocytidine into uridine. In support of this proposed scheme, a cytidine deaminase preparation obtained from E. coli catalyzed the decomposition of N4 aminocytidine into uridine and hydrazine. PMID- 3053654 TI - Chromosomal genes essential for stable maintenance of the mini-F plasmid in Escherichia coli. AB - We have isolated mutants of Escherichia coli which do not support stable maintenance of mini-F plasmids (delta ccd rep+ sop+). These host mutations, named hop, were classified into five linkage groups on the E. coli chromosome. Genetic analyses of these hop mutations by Hfr mating and P1 transduction showed their loci on the E. coli genetic map to be as follows: hopA in the gyrB-tnaA region, hopB in the bglB-oriC region, hopD between 8 and 15 min, and hopE in the argA thyA region. Kinetics of stability of the sop+ and delta sop mini-F plasmids in these hop mutants suggest that the hopA mutants are defective in partitioning of mini-F rather than in plasmid replication. The hopB, hopC, and hopD mutants were partially defective in replication of mini-F. The physical structure of the plasmid DNA was normal in hopA, B, C, and D mutants. Large amounts of linear multimers of plasmid DNA accumulated in mutants of the fifth linkage group (hopE). None of the hop mutations in any linkage group affected the normal growth of cells. PMID- 3053655 TI - Low-frequency infection of F- bacteria by transducing particles of filamentous bacteriophages. AB - Filamentous particles containing single-stranded plasmid and bacteriophage DNA are able to infect F- Escherichia coli at frequencies of approximately 10(-6). This infection is dependent on an intact particle and requires the products of the tolQ, tolR, and tolA genes of the bacteria. The addition of CaCl2 can increase the frequency about 100-fold, presumably by increasing the concentration of particles at the bacterial surface. PMID- 3053656 TI - Molecular cloning, expression, and analysis of the genes of the homoprotocatechuate catabolic pathway of Escherichia coli C. AB - The molecular cloning and fine-structure analysis of the homoprotocatechuate (hpc) catabolic pathway genes of Escherichia coli C are described. The genes were located in two operons, hpcBCDEF and hpcGH, that were very closely linked. A regulatory gene, hpcR, involved in the expression of both operons was also identified. Various subclones isolated in the study were useful in the production of chemical intermediates of the pathway. The availability of one such compound facilitated the discovery of a previously unrecognized isomerase involved in the catabolic sequence. PMID- 3053657 TI - Anaerobic regulation of pyruvate formate-lyase from Escherichia coli K-12. AB - The anaerobic regulation of the gene encoding pyruvate formate-lyase from Escherichia coli was investigated. Expression of a pfl'-'lacZ protein fusion demonstrated that the gene is subject to a 12-fold anaerobic induction which can be stimulated a further 2-fold by the addition of pyruvate to the growth medium. Construction of a strain deleted for pfl verified that either pyruvate or a metabolite of glycolysis functions as an inducer of pfl gene expression. Complete anaerobic induction required the presence of a functional fnr gene product. However, the dependence was not absolute since a two- to threefold anaerobic induction could still be observed in an fnr mutant. These results could be confirmed immunologically by analyzing the levels of pyruvate formate-lyase protein present in cells grown under various conditions. It was also shown that pfl'-'lacZ expression was partially repressed by nitrate and that this repression was mediated by the narL gene product. PMID- 3053658 TI - Cloning and expression in Escherichia coli of genes encoding a multiprotein complex involved in secretion of proteins from Staphylococcus aureus. AB - The genes encoding the multiprotein membrane-bound ribosomal protein (MBRP) complex (mrp genes), associated with membrane-bound ribosomes in Staphylococcus aureus, were cloned in Escherichia coli. All four components (molecular sizes 71, 60, 46, and 41 kilodaltons) of the MBRP complex were expressed from an 8.5 kilobase DNA fragment as judged by Western blot (immunoblot) analysis. The order of the individual genes within the cloned DNA fragment was determined by deletion mutagenesis and subcloning of various restriction fragments. Three RNAs, transcribed from the same DNA strand, were identified within the MBRP-coding region: one large RNA of approximately 5.9 kilobases, presumably coding for all four MBRP components, and two minor RNAs, coding for MBRP-71 and MBRP-60. The two minor RNAs seemed to be transcribed from promoters within the large transcription unit. Attempts to make insertional inactivations of the mrp genes with an internal 600-base-pair DNA fragment of the MBRP-coding region as a target were unsuccessful, presumably because such insertions are lethal. PMID- 3053659 TI - Molecular characterization of the tdc operon of Escherichia coli K-12. AB - The nucleotide sequence of a 2-kilobase DNA fragment of the tdc region of Escherichia coli K-12, previously cloned in this laboratory, revealed two open reading frames, tdcC and ORFX, downstream from the tdcB gene (formerly designated tdc) encoding biodegradative threonine dehydratase. A 24-base-pair sequence separated tdcC from the dehydratase coding region, and an untranslated region of 60 nucleotides, which contains a recognizable -10 consensus sequence, was found between tdcC and ORFX. The deduced amino acid sequence of tdcC showed it to be a large hydrophobic polypeptide of 431 amino acid residues, whereas ORFX coded for a small 135-residue polypeptide lacking glutamine and tryptophan. A computer assisted sequence analysis revealed no similarity among the tdcB, tdcC, and ORFX polypeptides, and a search of the GenBank database failed to detect similarity with any other known proteins. The tdc genes and ORFX showed similar codon usage and, in analogy with other bacterial genes, showed codon usage typical for genes expressed at an intermediate level. Transcriptional analysis with S1 nuclease indicated two distinct transcription start sites upstream of the tdcB gene in regions previously identified as promoterlike elements P1 and P2. Interestingly, expression of tdcB and tdcC, but not ORFX, was contingent upon the presence of P1. These results taken together tend to suggest that the biodegradative threonine dehydratase is the second gene in a polycistronic transcription unit constituting a novel operon (tdcABC) in E. coli implicated in anaerobic threonine metabolism. PMID- 3053660 TI - Genetic analysis of the tdcABC operon of Escherichia coli K-12. AB - The biodegradative threonine dehydratase (tdc) operon was mapped at 68 min on the Escherichia coli K-12 chromosome. The order of markers in the clockwise direction was dnaG uxaA tdc argG. A tdc deletion was isolated and mapped to this region of the chromosome. By using a tdcB-lacZ fusion the clockwise direction of transcription of tdc was determined. PMID- 3053662 TI - Relationship between low- and high-affinity glucose transport systems of Saccharomyces cerevisiae. AB - The high-affinity glucose transport process in Saccharomyces cerevisiae whole cells was regulated by catabolite repression and inactivation. The low-affinity process was constitutive, and its activity was inhibited in proportion to the extent of derepression of the high-affinity process. The latter finding suggests that there is some regulatory relationship between the two processes. PMID- 3053664 TI - Localization of the kdsA gene with the aid of the physical map of the Escherichia coli chromosome. AB - The isolation and analysis of two recombinant plasmids containing the kdsA gene from Escherichia coli chromosomal gene libraries is reported. The subfragments obtained from the inserts correspond to the fragment pattern around coordinate 1,282 kilobases of the physical map of the E. coli chromosome (Kohara et al. Cell 50:495-508, 1987). The kdsA gene has been located at coordinates 1,282 through 1,283 kilobases, corresponding to min 26.7 in the classical map coordinates. The kdsA gene is transcribed from this position toward the nearby nar gene. PMID- 3053661 TI - An in vivo complex with DNA photolyase blocks UV mutagenesis targeted at a thymine-cytosine dimer in Escherichia coli. AB - UV mutation frequency responses for two types of Escherichia coli prototrophic mutant were measured. Only the response associated with a mutation targeted by a thymine-cytosine pyrimidine dimer was reduced in the dark in cells with amplified DNA photolyase. This specific reduction is attributed to the interruption of mutational DNA synthesis by a photolyase complex at the targeting dimer. PMID- 3053663 TI - Conditionally lethal and recessive UGA-suppressor mutations in the prfB gene encoding peptide chain release factor 2 of Escherichia coli. AB - Strains carrying mutations in the prfB gene encoding peptide chain release factor 2 of Escherichia coli were isolated. prfB1, prfB2, and prfB3 were selected as suppressor mutations of a lacZ (UGA) mutation at 37 degrees C, one of which, prfB2, is temperature sensitive in growth. A prfB286 strain was selected as a conditionally lethal mutant which grows at 32 but not at 43 degrees C and was shown to have UGA-suppressor activity. All the mutations are recessive UGA suppressors. These data indicate that release factor 2 is essential to E. coli growth and that all mutants isolated here trigger suppression of the UGA codon. PMID- 3053666 TI - Glucose uptake in Saccharomyces cerevisiae grown under anaerobic conditions: effect of null mutations in the hexokinase and glucokinase structural genes. AB - Glucose uptake was investigated in a set of isogenic strains carrying a single glucose kinase structural gene, the other two kinase genes having been rendered nonfunctional through the construction of null mutations. Any one of the three kinases was sufficient for growth and glucose utilization aerobically or anaerobically. Under anaerobic conditions, substrate inhibition and regulation of carrier activity varied and depended upon the particular kinase present in the cell. PMID- 3053665 TI - Lipid modification of Escherichia coli penicillin-binding protein 3. AB - The primary structure of penicillin-binding protein 3 (PBP 3), an essential enzyme for cell division in Escherichia coli, was deduced from the nucleotide sequence of the ftsI gene (M. Nakamura, I. N. Maruyama, M. Soma, J. Kato, H. Suzuki, and Y. Hirota, Mol. Gen. Genet. 191:1-9, 1983). An amino acid sequence of Leu-26-Leu-Cys-Gly-Cys-30 was found near the amino terminus of the deduced sequence, showing a rather striking homology to the Leu-Leu-Ala-Gly-Cys consensus sequence for the modification and processing of precursors of the E. coli murein lipoprotein and other bacterial lipoproteins. As expected from this finding, PBP 3 was found to be modified with glycerol and fatty acids, although the lipid modification occurred only in a small fraction, accounting for less than 15% of the total PBP 3 molecules. PMID- 3053667 TI - mutM, a second mutator locus in Escherichia coli that generates G.C----T.A transversions. AB - We used strains carrying specific lacZ alleles to identify a new mutator locus in Escherichia coli which generates only G.C----T.A transversions among base substitutions. The locus, mutM, mapped near the cysE locus, which is at 81 min on the genetic map. PMID- 3053669 TI - Disulfiram-induced fulminating hepatitis: guidelines for liver-panel monitoring. AB - Although psychiatrists have medical responsibility for many alcoholic patients, the psychiatric literature, in contrast with the general medical literature, contains few reports of disulfiram-induced hepatotoxicity. For that reason, the authors review the literature on disulfiram hepatitis and report a case of severe fulminating hepatitis associated with disulfiram use, despite careful and currently accepted standard-of-care clinical and biochemical monitoring. All but two of the 17 disulfiram-associated hepatotoxic cases reviewed developed symptoms after 2 weeks to 2 months of use. Six patients died. This article discusses strategies for avoiding that rare but life-threatening side effect. The strategies include more frequent initial measurements of liver enzymes than is now accepted. Currently, only two reports recommend liver-function studies on a regular schedule for patients taking disulfiram. The authors believe that liver function tests should be administered before treatment, at 2-week intervals for 2 months, and at 3- to 6-month intervals thereafter. The authors emphasize that the hepatotoxicity reaction is rare and do not discourage the use of disulfiram in appropriate patients; rather, they wish to heighten the index of suspicion to disulfiram-induced hepatotoxicity. PMID- 3053668 TI - Fluoxetine versus trazodone in the treatment of outpatients with major depression. AB - Fluoxetine and trazodone were compared in a double-blind, randomized, parallel, 6 week trial in 43 outpatients with major depression after a 1-week single-blind placebo period. Thirty-five patients completed at least 3 weeks of active medication, while 25 patients completed all 6 weeks. Response rates, whether defined by end-of-treatment Hamilton Rating Scale for Depression (HAM-D) score less than 10 or by a 50% reduction in HAM-D scores, were equivalent for the two medications. For fluoxetine, HAM-D scores were significantly lower at Weeks 1 and 2 compared with those of trazodone. Trazodone improved sleep significantly more than fluoxetine. Fluoxetine was associated more frequently with weight loss (p = .002) and less frequently with dizziness (p = .04) than trazodone. PMID- 3053670 TI - ECT and antidepressants. PMID- 3053671 TI - ECT versus tricyclic antidepressants. PMID- 3053673 TI - Lithium efficacy and adverse effects. AB - The primary use of lithium is as maintenance treatment of bipolar disorders to prevent relapse. Although it effectively treats acute mania, lithium must often be used in conjunction with neuroleptics. Once-daily lithium therapy may help avoid some side effects and long-term renal damage. Neurotoxicity can be a problem for a small number of sensitive patients when neuroleptics and lithium are combined. Whenever possible, lithium should be avoided in the first trimester of pregnancy because of its potential for inducing cardiovascular abnormalities in the fetus. PMID- 3053672 TI - The use of anticonvulsants in manic-depressive illness. AB - The role of anticonvulsants, particularly carbamazepine, in the treatment of manic-depressive illness has been the subject of multiple recent studies. Overall, carbamazepine has achieved a 55% to 65% clinical response in mania. The drug has a time course and efficacy rate similar to those of neuroleptics and lithium. Studies have not uncovered a relationship between clinical response and carbamazepine levels in plasma or the CSF, but a relationship was found in one study between the drug's principal metabolite and clinical response. Patients who are severely depressed, manic, anxious, or dysphoric before treatment appear to be good responders to carbamazepine. In one trial, 40% of the manic patients taking carbamazepine experienced a marked response. In depression a moderate or better response to carbamazepine was attained by 50% of patients in one study. Some evidence indicates that carbamazepine can potentiate the efficacy of other medications, including lithium. PMID- 3053674 TI - Rat cytosolic aspartate aminotransferase: molecular cloning of cDNA and expression in Escherichia coli. AB - cDNA clones for rat cytosolic aspartate aminotransferase (cAspAT, L-aspartate:2 oxoglutarate aminotransferase) [EC 2.6.1.1] were isolated from a rat cDNA library, and the primary structure of the gene for cAspAT was deduced from its cDNA sequence. Rat cAspAT consists of 412 amino acids and its molecular weight is 46,295. The deduced amino acid sequence of rat cAspAT was compared with the sequences of AspATs from other species. The degree of sequence identities of rat/mouse cAspAT, rat/pig cAspAT, rat/chicken cAspAT, rat/pig mAspAT, and rat/Escherichia coli AspAT were 97.1, 89.6, 81.7, 48.1, and 41.2%, respectively. A coding region of rat cAspAT cDNA was inserted into E. coli expression vector pUC9, and enzymatically active cAspAT was expressed as a beta-galactosidase cAspAT hybrid protein. This hybrid protein represented about 18% of the soluble proteins in E. coli and its kinetic properties were comparable with those of cAspAT preparations purified from rat liver. PMID- 3053675 TI - Localization of paratropomyosin in skeletal muscle myofibrils and its translocation during postmortem storage of muscles. AB - Using polyclonal antibodies against paratropomyosin, which is believed to modify the actin-myosin interaction in postrigor skeletal muscles, we studied the localization of paratropomyosin in chicken breast muscle myofibrils. Intact myofibrils stained with fluorescent antibodies showed that paratropomyosin was exclusively located at the A-I junction region of sarcomeres. In stretched myofibrils (3.7 micron in sarcomere length), the approximate width of the fluorescent stripes and their relation to the A band remained constant. Removal of the A band from myofibrils led to loss of stainability. During postmortem storage of muscles, on the other hand, paratropomyosin was translocated from its original position at the A-I junction region onto thin filaments. The translocation of paratropomyosin was successfully induced with a calcium ion concentration of 10(-4) M in the presence of protease inhibitors. We therefore conclude that in postrigor muscles, paratropomyosin is released from the A-I junction region following the increase in the sarcoplasmic calcium ion concentration to 10(-4) M, and then binds to thin filaments, which results in weakening of rigor linkages formed between actin and myosin. PMID- 3053676 TI - Characterization of a monoclonal antibody against human placenta type IV collagen by immunoelectroblotting, antibody-coupled affinity chromatography, and immunohistochemical localization. AB - We have produced four monoclonal antibodies against type IV collagen obtained from human placenta. An antibody with a high titer by ELISA, named JK-199, reacted not only with type IV collagen in the triple-helical conformation but also with thermally denatured chains. After affinity chromatography on JK-199 antibody-coupled resin, the amino acid composition and CD spectrum of the affinity-purified peptides from the crude pepsin extract of human placenta were typical of those of human type IV collagen in the triple-helical conformation. On SDS-polyacrylamide gel electrophoresis, the purified protein showed only one broad band with a molecular weight of approximately 260,000 before reduction and six smaller peptide bands after reduction. On immunoelectroblotting, JK-199 reacted with all six peptide bands. Immunohistochemically, typical basement membranes were exclusively and strongly stained with JK-199 on frozen sections of PLP-fixed human placentas without any enzymatic pretreatment in the routine immunoperoxidase method. Judging from these findings, it is concluded that the epitopes of type IV collagen that reacted with JK-199 are exposed on the surface of basement membranes. This antibody should be useful for identification of type IV collagen in normal or pathological basement membranes or other structures. PMID- 3053677 TI - Expression of a rat liver microsomal cytochrome P-450 catalyzing testosterone 16 alpha-hydroxylation in Saccharomyces cerevisiae: vitamin D3 25-hydroxylase and testosterone 16 alpha-hydroxylase are distinct forms of cytochrome P-450. AB - Rat cytochrome P-450(M-1) cDNA was expressed in Saccharomyces cerevisiae TD1 cells by using a yeast-Escherichia coli shuttle vector consisting of P-450(M-1) cDNA, yeast alcohol dehydrogenase promoter and yeast cytochrome c terminator. The yeast cells synthesized up to 2 X 10(5) molecules of P-450(M-1) per cell. The microsomal fraction prepared from the transformed cells contained 0.1 nmol of cytochrome P-450 per mg of protein. The expressed cytochrome P-450 catalyzed 16 alpha- and 2 alpha-hydroxylations of testosterone in accordance with the catalytic activity of P-450(M-1), but did not hydroxylate vitamin D3 or 1 alpha hydroxycholecalciferol at the 25 position. The expressed cytochrome P-450 also catalyzed the oxidation of several drugs and did not show 25-hydroxylation activity toward 5 beta-cholestane-3 alpha, 7 alpha, 12 alpha-triol. However, it cross-reacted with the polyclonal and monoclonal antibodies elicited against purified P-450cc25 which catalyzed the 25-hydroxylation of vitamin D3. These results indicated that P-450(M-1) cDNA coded the 2 alpha- and 16 alpha hydroxylase of testosterone, and that these two positions of testosterone are hydroxylated by a single form of cytochrome P-450. Vitamin D3 25-hydroxylase and testosterone 16 alpha- and 2 alpha-hydroxylase are different gene products, although these two hydroxylase activities are immunochemically indistinguishable. PMID- 3053678 TI - Putative convertase involved in the proteolytic conversion of rat proalbumin to serum albumin. AB - We have found a proteolytic activity in Golgi membranes which efficiently converts [35S]methionine-labeled proalbumin, isolated from pulse-labeled rat hepatocytes in culture, to serum albumin in an in vitro assay system. The proalbumin-converting activity was dependent on Ca2+ and the maximum activity was observed at pH 5.5-6.0. Since the enzyme activity was found to be resistant not only to both leupeptin and E-64 but also to thiol-blocking reagents, it is unlikely that cathepsin B is involved in the proteolytic conversion of proalbumin occurring in the Golgi complex. PMID- 3053679 TI - Chemical modification of tryptophanase by chloramine T: a possible involvement of the methionine residue in enzyme activity. AB - Tryptophanase purified from Escherichia coli B/1t7-A was irreversibly inactivated by chloramine T (sodium N-chloro-p-toluenesulfonamide). The mode of inactivation was rather complex and did not follow pseudo-first-order kinetics. The inactivation of the apoenzyme was much faster than that of the holoenzyme. The Km value for the synthetic substrate S-o-nitrophenyl-L-cysteine (SOPC) increased concomitantly with the modification. In contrast, the Km value for the coenzyme, pyridoxal 5'-phosphate (PLP), was not altered. L-Serine, another substrate, and L alanine, a competitive inhibitor, protected the enzyme from inactivation. Determination of SH groups in the enzyme protein with 5,5'-dithiobis(2 nitrobenzoic acid) (DTNB) showed that modification of two SH groups per enzyme subunit resulted in a complete inactivation. When the enzyme was subjected to chloramine T-modification following the SH group modification with DTNB, further inactivation was still observed, even after the addition of dithiothreitol. The SH-blocked enzyme preparation thus obtained, however, exhibited less pH dependency of inactivation by chloramine T than that of the native enzyme. The amino acid analysis of the chloramine T-modified enzyme showed that modification of four or five methionine residues among the 16 residues per subunit proceeded concomitantly with the complete inactivation. Modification of the enzyme with chloramine T quenched the absorption peak near 500 nm, characteristic of a quinoidal structure formed by labilization of the alpha-proton. These results suggest the possibility that chloramine T modifies not only the SH groups, but also methionine residues important for the catalytic activity of the enzyme.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3053680 TI - A study on the identities of the three species of chromatin-associated proteinases in a mutant of Saccharomyces cerevisiae which lacks four major vacuolar proteinases. AB - Using mutant strain ABYS1 of Saccharomyces cerevisiae lacking four main vacuolar proteinases, proteinase A, proteinase B, carboxypeptidase Y, and carboxypeptidase S, we examined the identities of chromatin-associated proteinases, ruling out possible contamination of the chromatin fraction by them. The chromatin of strain ABYS1 showed three peaks of proteolytic activity at pH 4, 7, and 11, and these activities were found to be derived from three species of proteinases, the aspartic, serine neutral, and serine alkaline ones. As these chromatin-associated proteinases of strain ABYS1 were identical in both quality and quantity to those of wild-type strain of yeast, we suggest that the yeast chromatin contains three species of specific proteinases as essential components. PMID- 3053681 TI - Pseudomonas stutzeri ferredoxin: close similarity to Azotobacter vinelandii and Pseudomonas ovalis ferredoxins. AB - The complete primary structure of Pseudomonas stutzeri strain ZoBell ferredoxin was determined by a combination of protease digestion, Edman degradation, and carboxypeptidase digestion and was: TFVVTDNCIKCKYTDCVEVCPVDCFYEGPNFLVIH PDECIDCALCEPECPAQAIFSEDEVPEDQQEFIELNADLAEVWPNITE KKDALADAEEWDGVKDKLQYLER. The calculated molecular weight was 12,110 excluding iron and sulfur atoms. The amino acid sequence was highly homologous to those of Azotobacter vinelandii and Pseudomonas ovalis ferredoxins. It showed, like the other two, a Tyr-Thr insertion between the second and third Cys, and extra Cys at position 24 and, compared to Clostridium- and Bacillus-type ferredoxins, an extended C-terminal sequence. PMID- 3053683 TI - The intrinsic topological information of the wild-type and of up-promoter mutations of the Saccharomyces cerevisiae alcohol dehydrogenase II regulatory region. AB - A 569-base pair fragment encompassing the upstream regulatory region, the RNA initiation sites, and the initial part of the coding region of the Saccharomyces cerevisiae alcohol dehydrogenase II gene has been analyzed for the presence of sites which undergo conformational modification under torsional stress. Fine mapping of P1 and S1 endonuclease-sensitive sites was obtained on single topoisomers produced by in vitro ligation. It was shown that the upstream activator sequence, the TATA sequence, a region directly upstream to the RNA initiation sites, and several positions in the first segment of the transcribed region change conformation as a function of the applied torsional stress in a precisely coordinate fashion. The superhelical density optima for this coordinate modifications have been determined. Analysis of the conformational changes of the promoter sequence in several naturally occurring (Young, E. T., Williamson, V. M., Taguchi, A., Smith, M., Sledziewski, L., Russel, D., Osterman, J., Denis, C., Cox, D., and Beier, D., (1982) in Genetic Engineering of Microorganisms for Chemicals (Hollander, A., De Moss, R. D., Kaplan, S., Konisky, J., Savage, D., and Wolle, R. S., eds) pp. 335-361, Plenum Publishing Corp., New York) up promoter constitutive mutants was performed. This analysis has shown that the conformation of functionally relevant sites changes as a function of sequence mutations that have taken place elsewhere; this shows that the conformational behavior of the whole promoter region is linked and suggests transmission in cis of topological effects in RNA polymerase II promoters. PMID- 3053684 TI - The effects of pH on proton sugar symport activity of the lactose permease purified from Escherichia coli. AB - The lactose permease, which catalyzes galactoside-proton symport into Escherichia coli, has been purified and reconstituted in active form into artificial lipid vesicles. The roles of many detergents and phospholipids in solubilization and stabilization of the activity of the permease have been examined with a view to its eventual crystallization. Initial rates of uptake into reconstituted proteoliposomes determined by rapid mixing techniques proved that the activity of the permease can be comparable to that observed in the intact cell, while the best values for uptake rates obtained with conventional techniques were comparable to those reported for vesicles. The activity of the purified protein has been monitored over time periods of hours to weeks. It is shown that, under the best current conditions, the permease retains full activity for 1 to 2 weeks. Although this is still marginal for its crystallization, future improvements can now be assayed by rather stringent criteria. The mechanism of galactoside transport into reconstituted proteoliposome has been investigated by examining the effects of pH on influx into the vesicles. It is shown that the observed effects are entirely consistent with the predictions of a simple model of proton symport. The apparent increase in rate of uptake that is observed in the presence of a pH gradient is not so much due to an acceleration by a component of the protonmotive force as to the relaxation of inhibition by a product (internal protons) of the symport reaction. PMID- 3053685 TI - Topography of lactose permease from Escherichia coli. AB - The topography of lactose permease, in native membrane vesicles and after reconstitution of the purified protein into proteoliposomes, has been investigated by labeling the membrane-embedded portions of the protein using photoactivatable, hydrophobic reagents and by labeling the exposed portions of the protein with water-soluble, electrophilic reagents. Some sites of modification have been localized in fragments of the protein produced by chemical and enzymatic cleavage. These define a number of hydrophilic loops and membrane spanning regions and give some substance to topographic models of the permease. The N-terminal third of the molecule was labeled by three photoactivatable reagents (3-(trifluoromethyl)-3-m-iodophenyldiazirine and the phospholipid analogues 2-(aceto-(4-benzoylphenylether]-1-palmitoylphosphatidylcholine) and 2 (4-azido-2-nitrophenylaminoacetyl)-1-palmitoylphosphatidylcholin e) as well as the water soluble, electrophilic reagents. The C-terminal part of the molecule is labeled by the diazirine and, to a lesser extent, by the phospholipid analogues. It apparently has more nucleophilic groups accessible to water-soluble reagents than the N-terminal domain, in which the density of apparently unreactive ionizable residues proved to be unexpectedly high. The apparent lack of reactivity of some of these residues may be explained either by their being buried in the protein moiety within the membrane domain, or by their close association with other ionizable residues on the surface of the protein. PMID- 3053682 TI - Insulin-like growth factor I and insulin rapidly increase casein kinase II activity in BALB/c 3T3 fibroblasts. AB - We have tested whether growth factors added to serum-deprived BALB/c 3T3 fibroblasts alter the casein kinase II activity measured in cell extracts. A rapid phosphocellulose chromatography method was developed that provides a 40 fold partial purification of casein kinase II activity assayed with the specific substrate peptide Arg-Arg-Glu-Glu-Glu-Thr-Glu-Glu-Glu. Using this technique, kinase activity is stimulated 1.6-2.5-fold when isolated from fibroblasts treated with insulin or insulin-like growth factor I (IGF-I). The activated kinase activity exhibits the specific properties of casein kinase II such as the ability to utilize [gamma-32P]GTP as phosphate donor and marked inhibition by low concentrations of heparin. Activation of casein kinase II appears specific for these hormones because epidermal growth factor and platelet-derived growth factor have no effect on the kinase activity when added to fibroblasts under conditions where they markedly stimulate [3H]thymidine incorporation into DNA. Increases of casein kinase II activity by insulin and IGF-I were detected within 1 min of their addition to cell cultures. IGF-I is more potent in stimulating casein kinase II than insulin in mouse fibroblasts. These results demonstrate that casein kinase II is a selective target for insulin and IGF-I action in BALB/c fibroblasts, consistent with the hypothesis that this kinase plays a role in cellular signaling by these hormones. PMID- 3053686 TI - Primary structure of a human trifunctional enzyme. Isolation of a cDNA encoding methylenetetrahydrofolate dehydrogenase-methenyltetrahydrofolate cyclohydrolase formyltetrahydrofolate synthetase. AB - A DNA clone complementary to the messenger RNA encoding the human trifunctional enzyme 5,10-methylenetetrahydrofolate dehydrogenase-5,10-methenyl tetrahydrofolate cyclohydrolase-10-formyltetrahydrofolate synthetase has been isolated from a lambda gt10 library. In vitro transcription-translation of the 3.1-kilobase cDNA clone yields a protein of 101 kDa, which is of identical size and exhibits the same immunoreactivity as the enzyme purified from human liver. A coding region of 2805 base pairs in the cDNA encodes a protein of 935 amino acids. The initiator methionine is absent from the purified enzyme and the amino terminal 30 amino acids derived by automated sequence analysis are identical (arginine at position 18 was not identified) with that deduced from the nucleotide sequence. The amino acid sequence of the human enzyme shows extensive homology with that of the yeast enzyme, although the amino-terminal bifunctional dehydrogenase-cyclohydrolase domain is less homologous than the carboxyl-terminal synthetase domain. A region was identified which probably serves as a link between these two major domains of the human enzyme. The synthetase domain contains two regions that are homologous to consensus sequences for an ATP binding site. PMID- 3053687 TI - Site-specific alteration of cysteine 281, cysteine 344, and cysteine 349 in the proline carrier of Escherichia coli. AB - Cys-281, Cys-344, or Cys-349 in the proline carrier of Escherichia coli was changed to a serine residue by site-specific mutagenesis. The activities of the resultant mutants for uptake of proline were as great as that of the wild-type strain. These mutant carriers were all as sensitive as the wild-type carrier to the proline analogue azetidine 2-carboxylate. However, the mutant carriers with Ser-281 and Ser-344 were resistant to N-ethylmaleimide, whereas the mutant carrier with Ser-349 was as sensitive as the wild-type carrier to this reagent. These results indicate that these cysteine residues are not essential for proline transport and that Cys-281 and Cys-344 may be close to the substrate-binding site that contains an N-ethylmaleimide-sensitive residue. PMID- 3053688 TI - Roles of the narJ and narI gene products in the expression of nitrate reductase in Escherichia coli. AB - Nitrate reductase, released and purified from membrane fractions of Escherichia coli, is composed of three subunits. Formation of the enzyme depends on induction of the nar operon, narGHJI, which is composed of four open reading frames (ORF). Previous studies established that the first two genes in the operon narG and narH encode the alpha and beta subunits, respectively, while formation of the gamma subunit, cytochrome bNR, depends on expression of the promoter distal genes. The narJ and narI genes were subcloned separately into plasmids where each was under the control of the nar promoter. Expression of these plasmids in a mutant which forms only alpha and beta subunits revealed that expression of the narI gene is sufficient to restore normal levels of cytochrome bNR, but expression of both genes is required for assembly of fully active, membrane-bound nitrate reductase. The amino acid composition, the N-terminal sequence, and the sequence of cyanogen bromide fragments derived from the isolated gamma subunit corresponds to that expected for a protein produced by the narI ORF. A protein corresponding to the narJ ORF did not appear to be associated with the purified nitrate reductase complex or with the complex immunoprecipitated from Triton X-100-solubilized membrane preparations. We conclude that narI encodes the gamma subunit (cytochrome bNR) and that while the product of the narJ gene is required for assembly of fully active membrane-bound enzyme it is not tightly associated with the active enzyme. PMID- 3053689 TI - Production and characterization of human basic fibroblast growth factor from Escherichia coli. AB - The cDNA sequence coding for human basic fibroblast growth factor (bFGF) has been cloned downstream of a transcription promoter recognized by the bacteriophage T7 RNA polymerase. Initiation of translation at the fgf gene has been coupled to upstream translation of a fragment of the T7 phi 10 gene. Expression of the fgf gene in this system can lead to an accumulation of approximately 40 mg/liter/A600 unit of bFGF. This material can be purified close to homogeneity from a soluble protein extract on a heparin-Sepharose column. bFGF so obtained has been shown to have bioactivity indistinguishable from human placental fibroblast growth factor in mitogenicity, synthesis of plasminogen activator, and angiogenesis assays. PMID- 3053690 TI - Prohormone processing and the secretory pathway. PMID- 3053692 TI - Cleavage of type I and type II procollagens by type I/II procollagen N proteinase. Correlation of kinetic constants with the predicted conformations of procollagen substrates. AB - The kinetic constants were examined for the cleavage of several types of procollagen by type I/II procollagen N-proteinase. The Km values were essentially the same (0.2 microM) for chick type I procollagen, human type I procollagen, and chick type II procollagen. However, the Vmax values differed over a 14-fold range. As reported previously, the enzyme did not cleave denatured type I or II procollagen. Also, it did not cleave human type III procollagen which contains the same scissle -Pro-Gln- bond as the pro-alpha 1(I) chain of type I procollagen. To explain the observations, Chou-Fasman rules were used to compare the secondary structures of the cleavage sites in the procollagens. The results supported a previous suggestion (Helseth, D. L., Jr., Lechner, J. L., and Veis, A. (1979) Biopolymers 18, 3005-3014) that the region carboxyl-terminal to cleavage site in the pro-alpha 1(I) chain of type I procollagen was in a hairpin conformation consisting of a beta-sheet, beta-turn, and beta-sheet. In both chick and human type I procollagen, the hairpin loop in the pro-alpha 1(I) chain consisted of about 18 amino acids. The cleavage site itself was in a short alpha helical structure of four or five amino acids. The pro-alpha 2(I) chains had a similar hairpin loop of about 14 amino acids and alpha-helix of four or five amino acids containing the cleavage site. Chick type II procollagen, which had the highest Vmax value, had a longer hairpin structure of 22 amino acids, and the cleavage site was in a longer alpha-helical domain of 10 amino acids. In contrast, type III procollagen had a random-coil conformation in the same region. The results help to explain the unusual substrate requirements of type I/II N proteinase. They also help explain why mutations that produce in-frame deletions of amino acids 84 or more residues carboxyl-terminal to the cleavage site make the protein resistant to the enzyme. PMID- 3053691 TI - Evidence for interaction of an aminoacyl transfer RNA synthetase with a region important for the identity of its cognate transfer RNA. AB - Recent experiments showed that a single base pair (G3:U70) in the amino acid acceptor helix is a major determinant for the identity of Escherichia coli alanine transfer RNA. Experiments reported here show that bound alanine tRNA synthetase protects (from ribonuclease attack) seven consecutive phosphodiester linkages on the 3'-side of the acceptor-T psi C helix (phosphates 65-71) and a few additional sites that are in scattered locations. There is no evidence for interaction of the enzyme with the anticodon, a sequence which can be varied without effect on recognition by alanine tRNA synthetase. PMID- 3053693 TI - Analysis of sequential steps of nucleotide excision repair in Escherichia coli using synthetic substrates containing single psoralen adducts. AB - Escherichia coli ABC excinuclease initiates the removal of dodecanucleotides from damaged DNA in an ATP-dependent reaction. Using a synthetic DNA fragment containing a psoralen adduct at a defined position we have investigated the interaction of the components of the enzyme with substrate by DNase I footprinting. We find that the UvrA subunit binds to DNA specifically in the absence of cofactors and that the binding affinity is stimulated about 4-fold by ATP and only marginally inhibited by ADP. The UvrA.DNA complexes formed in the absence of co-factors or in the presence of either ATP or ADP are remarkably similar. In contrast, adenosine 5'-O-(thiotriphosphate) increases nonspecific binding and completely abolishes the UvrA footprint. The UvrB subunit can associate with the UvrA subunit on DNA in the absence of ATP, but this ternary UvrA.UvrB.DNA complex is qualitatively different from that formed in the presence of ATP. The UvrC subunit elicits no additional change in the UvrA-UvrB footprint. Helicase II (UvrD protein) does not alter the UvrA-UvrB footprint but does appear to interact at the 5'-incision site of the postincision complex. DNA polymerase I fills in the excision gap in the presence or absence of helicase II and apparently releases the ABC excinuclease from the repaired DNA. Nearly 90% of the repair patches are 12 nucleotides long, and this length is not affected by helicase II. We see no evidence by DNase I footprinting for the formation of a multiprotein complex encompassing the UvrA, -B, -C, and -D proteins and DNA polymerase I. PMID- 3053695 TI - Gastrin gene expression and regulation in rat islet cell lines. AB - Gastrin gene expression was observed in two permanent rat insulinoma (RIN) cell lines derived from a rat insulinoma. Gastrin expression was selective; highest expression was seen in a cell line which did not express other islet cell hormones. Gastrin mRNA transcription initiated from the same promoter as antral gastrin mRNA. DNA transfection studies with a gastrin chloramphenicol acetyltransferase chimeric gene showed higher expression in gastrin-expressing RIN cells than non-gastrin-expressing islet cells. This implies that gastrin expressing RIN cells selectively express a trans-acting transcriptional activator which binds to cis-acting regulatory sequences within the 5'-flanking DNA sequence and first exon of the gastrin gene. The gastrin peptide precursor synthesized in these RIN cell lines is subject to the same repertoire of posttranslational modifications within the cell's secretory apparatus (endoproteolytic cleavage, tyrosine sulfation, and C-terminal amidation) as seen in antral G cells. Gastrin mRNA levels in these RIN cells were selectively increased by increasing the extracellular calcium concentration. Membrane depolarization also stimulated gastrin mRNA levels, probably through activation of voltage-sensitive calcium channels. Thus, these gastrin-expressing RIN cell lines provide permanent cell lines useful in analyzing the cellular regulation of gastrin gene expression. PMID- 3053694 TI - The alpha-lytic protease gene of Lysobacter enzymogenes. The nucleotide sequence predicts a large prepro-peptide with homology to pro-peptides of other chymotrypsin-like enzymes. AB - alpha-Lytic protease is a 19.8-kDa protein secreted from the Gram-negative bacterium Lysobacter enzymogenes. We have cloned and sequenced the gene for this serine protease. The nucleotide sequence contains an open reading frame which codes for the 198-residue mature enzyme and a potential prepro-peptide, also of 198 residues. The COOH-terminal 49 residues of the pro-peptide are significantly homologous to the propeptides of Streptomyces griseus proteases A and B. We suggest that this pro-peptide region facilitates formation of the active enzyme. A region bridging the NH2-terminal pre- and pro-peptides is homologous to a maize inhibitor of serine proteases. We speculate that this region inhibits enzymatic activity of the prepro-enzyme. PMID- 3053696 TI - Occurrence of 9-deoxy-delta 9,delta 12-13,14-dihydroprostaglandin D2 in human urine. AB - We have developed a highly sensitive and specific solid-phase enzyme immunoassay for 9-deoxy-delta 9,delta 12-dihydroprostaglandin D2 (delta 12-PGJ2) and studied the occurrence of this novel PGD2 metabolite in human urine. The assay detected delta 12-PGJ2 over the range of 2-200 pg, and the antiserum showed 2% cross reaction with PGJ2 and less than 0.2% with other PGs. We used this assay and purified the delta 12-PGJ2-like immunoreactive substance from human urine. Purification consisted of chromatographies on a Sep-Pak C18 cartridge, a silicic acid column, reversed-phase high-performance liquid chromatography, and finally an affinity column of anti-delta 12-PGJ2 antibody. As a result, about 850 ng of delta 12-PGJ2-like immunoreactive substance were recovered from 60 liters of human urine. The purified material was identified as delta 12-PGJ2 by gas chromatography/high resolution-selected ion monitoring using the molecular ion m/z 448[M]+. and ions [M - 15]+, [M - 43]+, [M - 100]+., and [M - 143]+. The amounts of delta 12-PGJ2 in the urine from normal, volunteer men and women were 151.5 +/- 20.0 and 65.6 +/- 5.4 ng/24 h (mean +/- S.E., n = 5), respectively. The delta 12-PGJ2 amount in urine did not alter significantly during storage for at least 24 h or by the addition of authentic PGD2 to urine samples, suggesting that the delta 12-PGJ2 we determined was not derived from the decomposition of PGD2 in the urine during storage or purification. Moreover, when a single dose of PGD2 (1 mg/kg) was injected intravenously into cynomolgus monkeys, the urinary level of delta 12-PGJ2 increased 20- to 180-fold over the normal levels, whereas the delta 12-PGJ2 level decreased by 40-50% of the normal levels, following the administration of indomethacin at a dose of 1 mg/kg. These results indicate that delta 12-PGJ2 is formed naturally in the body and excreted as a urinary PGD2 metabolite. PMID- 3053697 TI - Primary structure of the lactose permease gene from the yeast Kluyveromyces lactis. Presence of an unusual transcript structure. AB - The LAC12 gene of Kluyveromyces lactis codes for an inducible lactose permease. We have determined the nucleotide sequence of a DNA fragment which includes the complete LAC12 gene. The 4.7-kilobase (kb) mRNA carrying LAC12 contained two open reading frames, ORFI (1761 bases) and ORFII (1266 bases), separated by a 573-base pair noncoding region. Mung bean and exonuclease VII mapping showed that there was no splicing of the 4.7-kb transcript and thus no intron between the two open reading frames. Chromosomal disruption of ORFI with the URA3 gene destroyed lactose transport activity, suggesting that ORFI codes for a component of the permease. Disruption of ORFII and the noncoding region between the two open reading frames did not affect the lactose permease function, indicating that they do not comprise a part of the permease. We do not know if ORFII is translated, but in either case, the structure of the 4.7-kb mRNA is unusual. We discuss possible origins for it. The peptide predicted from ORFI is hydrophobic as would be expected for a membrane-bound protein. Compared with other membrane proteins, LAC12 (ORFI) protein showed sequence similarity to the human glucose and the Escherichia coli xylose-H+ and arabinose-H+ transporters. No obvious amino acid sequence similarity was found with the lactose permease of E. coli. PMID- 3053698 TI - Deletion of the amino-terminal domain of asialoglycoprotein receptor H1 allows cleavage of the internal signal sequence. AB - Human asialoglycoprotein receptor H1 is a single-spanning membrane protein with an amino-terminal domain of 40 residues exposed to the cytoplasm and the carboxyl terminal domain translocated to the exoplasmic side of the membrane. It has been shown earlier that the transmembrane segment functions as an internal uncleaved signal sequence for insertion into the endoplasmic reticulum. In a deletion protein lacking almost the entire cytoplasmic domain, the signal sequence is cleaved at the carboxyl-terminal end of the transmembrane segment. All available criteria suggest that the protein is processed by signal peptidase. The cytoplasmic domain of the receptor does not directly inhibit signal cleavage since it does not detectably hinder cleavage of the normally amino-terminal signal sequence of influenza hemagglutinin in fusion proteins. We suggest that by its size or structure it affects the position of the receptor in the membrane and thus the accessibility of the potential cleavage site to signal peptidase. PMID- 3053699 TI - D-serine dehydratase from Escherichia coli. DNA sequence and identification of catalytically inactive glycine to aspartic acid variants. AB - We have identified two glycyl residues whose integrity is essential for the catalytic competence of a model pyridoxal 5'-phosphate requiring enzyme, D-serine dehydratase from Escherichia coli. This was accomplished by isolating and sequencing the structural gene from wild type E. coli and from two mutant strains that produce inactive D-serine dehydratase. DNA sequencing indicated the presence of a single glycine to aspartic acid replacement in each variant. The amino acid replacements lie in a glycine-rich region of D-serine dehydratase well removed from pyridoxal 5'-phosphate-binding lysine 118 in the primary structure of the enzyme. The striking effect of these two glycine to aspartic acid replacements on catalytic activity, the conservation of the glycine-rich region in several pyridoxal 5'-phosphate-dependent enzymes that catalyze alpha/beta-eliminations, and the placement of similar glycine-rich sequences in well-characterized active site structures suggest that the glycine-rich region interacts with the cofactor at the active site of the enzyme. PMID- 3053700 TI - The glycine-rich region of Escherichia coli D-serine dehydratase. Altered interactions with pyridoxal 5'-phosphate produced by substitution of aspartic acid for glycine. AB - Replacement of glycine by aspartic acid at either of two sites in a conserved, glycine-rich region inactivates the pyridoxal 5'-phosphate-dependent enzyme D serine dehydratase (DSD) from Escherichia coli. To investigate why aspartic acid at position 279 or 281 causes a loss of activity, we measured the affinity of the G----D variants for pyridoxal 5'-phosphate and a cofactor:substrate analog complex and compared the UV, CD, and fluorescence properties of wild-type D serine dehydratase and the inactive variants. The two G----D variants DSD(G279D) and DSD (G281D) displayed marked differences from wild-type D-serine dehydratase and from each other with respect to their affinity for pyridoxal 5'-phosphate and for a pyridoxal 5'-phosphate:glycine Schiff base. Compared to the wild-type enzyme, the cofactor affinity of DSD(G279D) and DSD(G281D) was decreased 225- and 50-fold, respectively, and the ability to retain a cofactor:glycine complex was decreased 765- and 1970-fold. The spectral properties of the inactive variants suggest that they form a Schiff base linkage with pyridoxal 5'-phosphate but do not hold the cofactor in a catalytically competent orientation. Moreover, the amount of cofactor aldamine in equilibrium with cofactor Schiff base is increased in DSD(G279D) and DSD(G281D) relative to that in wild-type DSD. Collectively, our findings indicate that introduction of a carboxymethyl side chain at G-279 or G 281 directly or indirectly disrupts catalytically essential protein-cofactor and protein-substrate interactions and thereby prevents processing of the enzyme bound cofactor:substrate complex. The conserved glycine-rich region is thus either an integral part of the D-serine dehydratase active site or conformationally linked to it. PMID- 3053701 TI - Mechanism of stimulation of T7 DNA polymerase by Escherichia coli single-stranded DNA binding protein (SSB). AB - Single-stranded DNA binding protein is a key component in growth of bacteriophage T7. In addition, DNA synthesis by the purified in vitro replication system is markedly stimulated when the DNA template is coated with Escherichia coli single stranded DNA binding protein (SSB). In an attempt to understand the mechanism for this stimulation, we have studied the effect of E. coli SSB on DNA synthesis by the T7 DNA polymerase using a primed single-stranded M13 DNA template which serves as a model for T7 lagging strand DNA synthesis. Polyacrylamide gel analysis of the DNA product synthesized on this template in the absence of SSB indicated that the T7 DNA polymerase pauses at many specific sites, some stronger than others. By comparing the position of pausing with the DNA sequence of this region and by using a DNA template that contains an extremely stable hairpin structure, it was found that many, but not all, of these pause positions correspond to regions of potential secondary structure. The presence of SSB during synthesis resulted in a large reduction in the frequency of hesitations at many sites that correspond to these secondary structures. However, the facts that a large percentage of the pause sites remain unaffected even at saturating levels of SSB and that SSB stimulates synthesis on a singly primed poly(dA) template suggested that other mechanisms also contribute to the stimulation of DNA synthesis caused by SSB. Using a sucrose gradient analysis, we found that SSB increases the affinity of the polymerase for single-stranded DNA that this increased binding is only noticed when the polymerase concentration is limiting. The effect of this difference in polymerase affinity was clearly observed by a polyacrylamide gel analysis of the product DNA synthesized during a limited DNA synthesis reaction using conditions where only two nucleotides are added to the primer. Under these circumstances, where the presence of hairpin structures should not contribute to the stimulatory effect of SSB, we found that the extension of the primer is stimulated 4-fold if the DNA template is coated with SSB. Furthermore, SSB had no effect on this synthesis at large polymerase to template ratios. PMID- 3053702 TI - cDNA-derived primary structure of the glycoprotein component of canine microsomal signal peptidase complex. AB - Canine microsomal signal peptidase activity has been shown previously to co migrate as an apparent complex of six polypeptides with molecular masses of 25, 23, 22, 21, 18, and 12 kDa. The 22- and 23-kDa species are differentially glycosylated forms of the same protein, designated SPC 22/23. The amino acid sequence of SPC 22/23 was deduced from cDNA clones. The protein is synthesized without a cleavable amino-terminal signal sequence and contains a single site for N-linked glycosylation. SPC 22/23 appears to be anchored to the rough endoplasmic reticulum membrane by a single hydrophobic segment near its amino terminus, with the remainder of the protein positioned on the lumenal side of the membrane. The amino acid sequence of SPC 22/23 shares homology with tryptic peptides derived from the hen oviduct signal peptidase glycoprotein, one of two possible proteins required for signal peptide processing in the avian system (Baker, R.K., and Lively, M.O. (1987) Biochemistry 26, 8561-8567). Therefore, the complete amino acid sequence of SPC 22/23 presented in this report corresponds to one of two possible proteins required for signal peptide processing in higher eukaryotic cells. PMID- 3053703 TI - Glutathione metabolism and its selective modification. PMID- 3053704 TI - Site-directed mutagenesis of the tryptophan residues in yeast eukaryotic initiation factor 4E. Effects on cap binding activity. AB - Initiation factor 4E is a 24-kilodalton polypeptide that binds specifically to the 5' cap structure of eukaryotic mRNAs. Sequence analysis of cDNA clones of initiation factor 4E from several species revealed a high tryptophan content (8 residues). Strikingly, all tryptophans are conserved evolutionarily in number and position between yeast and mammals. Here we show, using site-directed mutagenesis, that two of the tryptophans (those referred to as numbers 1 and 8) are absolutely required for the cap binding activity of an Escherichia coli expressed initiation factor 4E. PMID- 3053705 TI - An islet amyloid peptide is derived from an 89-amino acid precursor by proteolytic processing. AB - Amyloid deposits occurring in the islets of Langerhans in patients with noninsulin-dependent diabetes mellitus and some insulinomas contain a 37-amino acid peptide that is structurally related to calcitonin gene-related peptide. We have identified three cDNA clones encoding islet amyloid polypeptide (IAPP) or diabetes-associated peptide (DAP) by oligonucleotide screening of a lambda gt10 human insulinoma cDNA library. Two of the three cDNAs contained a domain encoding IAPP/DAP but had an intron-like sequence in their 5' region. The other cDNA contained an open reading frame encoding an 89-amino acid precursor having a typical signal peptide followed by a small prohormone-like sequence containing within it the IAPP/DAP peptide bracketed at its NH2 and COOH termini by Lys-Arg and Gly-Lys-Arg, respectively. These data indicate that this amyloid peptide is generated by proteolytic processing similar to that for proinsulin and other islet prohormones and also that the peptide may be carboxyamidated. The isolation of cDNA clones having 5'-unprocessed intron-like sequences suggests that inefficient or alternative splicing of this mRNA occurred in the insulinoma. PMID- 3053706 TI - Purification and characterization of proteins that bind to yeast ARSs. AB - Two proteins that bind to yeast ARS DNA have been purified using conventional and oligonucleotide affinity chromatography. One protein has been purified to homogeneity and has a mass of 135 kDa. Competitive binding studies and DNase I footprinting show that the protein binds to a sequence about 80 base pairs away from the core consensus in the region known as domain B. This region has previously been shown to be required for efficient replication of plasmids carrying ARS1 elements. To investigate further whether the protein might have a function related to the ability of ARSs to act as replicators, binding to another ARS was tested. The protein binds to the functional ARS adjacent to the silent mating type locus HMR, called the HMR-E ARS, about 60 base pairs from the core consensus sequence. Surprisingly, there is little homology between the binding site at the HMR-E ARS and the binding site at ARS1. The 135-kDa protein is probably the same as ABF-I (SBF I) (Shore, D., Stillman, D. J. Brand, A. H., and Nasmyth, K. A. (1987) EMBO J. 6, 461-467; Buchman, A. R., Kimmerly, W. J., Rine, J., and Kornberg, R. D. (1988) Mol. Cell. Biol. 8, 210-225). A second DNA-binding protein was separated from ABF-I during later stages of the purification. This protein, which we designate ABF-III, also binds specifically to the ARS1 sequence, as shown by DNase I footprinting, at a site adjacent to the ABF-I recognition site. Purification of these two ARS binding proteins should aid in our understanding of the complex mechanisms that regulate eukaryotic DNA replication. PMID- 3053707 TI - Selective inhibition of glycoprotein-processing enzymes. Differential inhibition of glucosidases I and II in cell culture. AB - In this study, we compared the effects of 2,6-dideoxy-2,6-imino-7-O-(beta-D glucopyranosyl)-D-glycero-L-gulohep titol (MDL) to those of the glucosidase I inhibitor, castanospermine, on the purified processing enzymes glucosidases I and II. WE also compared the effects of these two inhibitors on glycoprotein processing in cell culture using influenza virus-infected Madin-Darby canine kidney cells as a model system. With the purified processing enzymes, castanospermine was a better inhibitor of glucosidase I than of glucosidase II, whereas MDL is more effective against glucosidase II than glucosidase I. In cell culture at the appropriate dose, MDL also preferentially affected glucosidase II. Thus, at 250 micrograms/ml MDL, the major [3H]glucose-labeled (or [3H]mannose labeled) glycopeptide from the viral hemagglutinin was susceptible to endoglucosaminidase H, and the oligosaccharide liberated by this treatment was characterized as a Glc2Man7-9GlcNAc on the basis of size, resistance to digestion by glucosidase I (but sensitivity to glucosidase II), methylation analysis, and Smith degradation studies. These data indicate that at appropriate concentrations of MDL (250 micrograms/ml), one can selectively inhibit glucosidase II in Madin Darby canine kidney cells. However, at higher concentrations of inhibitor (500 micrograms/ml), both enzymes are apparently affected. Since MDL did not greatly inhibit the synthesis of lipid-linked saccharides or the synthesis of protein or RNA, it should be a useful tool for studies on the biosynthesis and role of N linked oligosaccharides in glycoprotein function. PMID- 3053708 TI - Evidence for a role of protein kinase C in luteinizing hormone synthesis and secretion. Impaired responses to gonadotropin-releasing hormone in protein kinase C-depleted pituitary cells. AB - The role of protein kinase C in luteinizing hormone (LH) release was analyzed in studies on the actions of phorbol esters and gonadotropin-releasing hormone (GnRH) in normal and protein kinase C (Ca2+/phospholipid-dependent enzyme) depleted pituitary cell cultures. LH secretory responses of normal pituitary cells to GnRH were reduced but not abolished in Ca2+-deficient medium, consistent with the existence of extracellular Ca2+-dependent and -independent components of GnRH action. Both of these components could be elicited by treatment with 12-O tetradecanoylphorbol 13-acetate (TPA). The LH secretory responses to TPA and GnRH were additive only at low doses and converged to a common maximum at high concentrations of the agonists in the presence or absence of extracellular Ca2+. The release of stored LH by GnRH and TPA was accompanied by secretion of newly synthesized LH from 2 to 5 h during stimulation by either of the agonists. LH synthesis was increased in a progressive and dose-dependent manner by GnRH and TPA, and the ratio between newly synthesized and released hormone was near 1:2. TPA caused rapid and complete translocation of cytosolic protein kinase C to the particulate fraction of pituitary cells, followed by a progressive decrease in total enzyme content to approximately 10% after 6 h. Partial recovery of the cytosolic enzyme (to 20%) occurred after washing and reincubation for 15 h. Such kinase C-depleted cells showed prominent, dose-dependent reductions in the actions of GnRH and TPA on LH release and synthesis in both normal and Ca2+ deficient media. These observations support the hypothesis that protein kinase C participates in LH biosynthesis and secretion in pituitary gonadotrophs and is involved in the actions of GnRH upon these processes. PMID- 3053710 TI - Characterization of dolichol diphosphate oligosaccharide: protein oligosaccharyltransferase and glycoprotein-processing glucosidases occurring in trypanosomatid protozoa. AB - We have previously reported that the oligosaccharides transferred in vivo from dolichol-P-P derivatives in protein N-glycosylation in trypanosomatids are devoid of glucose residues and contain 2 N-acetylglucosamine and 6, 7, or 9 mannose units depending on the species. In this respect trypanosomatids differ from wild type mammalian, plant, insect, and fungal cells in which Glc3Man9GlcNAc2 is transferred. We are now reporting that incubation of Glc1-3Man9GlcNAc2-P-P dolichol and Man7-9GlcNAc2-P-P-dolichol with membranes of Trypanosoma cruzi, Leptomonas samueli, Crithidia fasciculata, and Blastocrithidia culicis and an acceptor hexapeptide leads to the transfer of the six above mentioned lipid linked oligosaccharides at the same rate. Control experiments performed under similar conditions but with rat liver and Saccharomyces cerevisiae membranes showed that, as already known, Glc3Man9GlcNAc2 is preferentially transferred in the latter systems. We have also previously reported that, once transferred to protein, the oligosaccharides become transiently glucosylated in trypanosomatids. Depending on the species, protein-linked Glc1Man5-9GlcNAc2 have been transiently detected in cells incubated with [14C] glucose. We are now reporting that glucosidase activities degrading both Glc1Man9GlcNAc2 and Glc2Man9GlcNAc2 were detected in T. cruzi, L. samueli, and C. fasciculata. The enzymatic activities were associated with a membrane fraction; they had a neutral optimum pH value, and similarly to mammalian glucosidase II, the enzyme acting on the monoglucosylated substrate showed a decreased affinity when the latter contained fewer mannose residues. No glucosidase I-like enzyme acting on Glc3Man9GlcNAc2 was detected in any of the three above-mentioned protozoan species. This result is consistent with the fact that no oligosaccharides containing 3 glucose units occur in trypanosomatids. PMID- 3053709 TI - Mechanism of androstenedione formation from testosterone and epitestosterone catalyzed by purified cytochrome P-450b. AB - A purified rat hepatic monooxygenase system containing cytochrome P-450b oxidizes testosterone to androstenedione and 16 alpha- and 16 beta-hydroxytestosterone at approximately equal rates. The metabolism of epitestosterone by the same system is characterized by a marked stereoselectivity in favor of 16 beta-hydroxylation (4- to 5-fold relative to 16 alpha-hydroxylation), formation of 15 alpha hydroxyepitestosterone, and a rate of androstenedione formation which is three to five times higher than that observed with testosterone. Apparent Km values for 16 alpha- and 16 beta-hydroxylation and androstenedione formation are 20-30 microM with either substrate. Mass spectral analysis of the androstenedione formed from [16,16-2H2]testosterone and [16,16-2H2] epitestosterone indicates essentially complete retention of deuterium, thereby ruling out a mechanism of androstenedione formation via C-16 hydroxylation followed by loss of water and rearrangement. Mass spectral analysis of the C-16 hydroxylation products from incubations of testosterone or epitestosterone in 18O2 shows essentially complete incorporation of 18O (greater than 95%). Androstenedione formed from testosterone is enriched in 18O only 2-fold (5-8%) over background, while the androstenedione formed from epitestosterone shows 84% enrichment. Kinetic experiments utilizing [17-2H]testosterone and [17-2H]epitestosterone as substrates indicate that cleavage of the C-17 carbon-hydrogen bond is involved in a rate-limiting step in the formation of androstenedione from both substrates. Taken together, our results indicate that androstenedione formation from epitestosterone proceeds exclusively through the gem-diol pathway, while androstenedione formation from testosterone may proceed through a combination of gem-diol and dual hydrogen abstraction pathways. PMID- 3053711 TI - Studies of RNA-protein interactions in the yeast 5 S ribonucleoprotein particles by fluorescence and tritium exchange. Implications for ribosomal assembly. AB - Studies of the conformational properties of the yeast 5 S RNA-protein complex were initiated in an attempt to understand loss of ability of its individual protein and RNA components to reassociate. The 5 S RNA-L1a protein complex from 60 S ribosomal subunits of Saccharomyces cerevisiae could be dissociated by high concentrations of magnesium. The degree of dissociation could be monitored by polyacrylamide gel electrophoresis. The complex was completely dissociated at about 390 mM magnesium, but was stable at 4 degrees C in 25 mM EDTA up to 48 h. The overall conformation of the complex was monitored using tritium exchange. The tritium exchange behavior was dramatically changed as the complex was dissociated. To determine contribution of each component to the observed overall change reflected in the tritium exchange behavior, ethidium bromide (EtBr) and bis-anilinonaphthalene-sulfonic acid fluorescence were used to monitor the RNA and the protein moiety, respectively. Upon dissociation of the complex, the fluorescence intensity resulting from EtBr binding to RNA decreased, whereas the intensity due to bis-anilinonaphthalene-sulfonic acid binding to the protein increased. Turbidity was observed during dissociation of the complex. These results indicate that disruption of interactions between the 5 S RNA and protein L1a resulted in an exposure of solvent-accessible apolar regions in the protein molecule. Such exposure led to insolubility of protein and irreversibility in interaction between individual components. Properties of the separated components also suggest that special conditions may be required for these components to associate during ribosomal assembly. PMID- 3053712 TI - The factor V-activating enzyme (RVV-V) from Russell's viper venom. Identification of isoproteins RVV-V alpha, -V beta, and -V gamma and their complete amino acid sequences. AB - The complete amino acid sequences of two isoproteins of the factor V-activating enzyme (RVV-V) isolated from Vipera russelli (Russell's viper) venom were determined by sequencing S-pyridylethylated derivatives of the proteins and their peptide fragments generated by either chemical (cyanogen bromide and 2-(2 nitrophenylsulfenyl)-3-methyl-3-bromoindolenine) or enzymatic (trypsin, alpha chymotrypsin, and lysyl endopeptidase) cleavages. Both enzymes, designated RVV-V alpha and RVV-V gamma, consist of 236 amino acid residues and have a N-linked oligosaccharide chain at Asn229. The six amino acid substitutions between RVV-V alpha and -V gamma are: Thr22(alpha)-Ala22(gamma), Gly29(alpha)-Ala29(gamma), Gln191(alpha)-Glu191(gamma), Ile192(alpha)-Met192(gamma), Gln193(alpha) His193(gamma), and Asn224(alpha)-Ser224(gamma). The molecular weights were calculated as 26,182 for RVV-V alpha and 26,167 for RVV-V gamma. The sequences of the RVV-V isoproteins exhibited 62% identity with that of batroxobin, a thrombin like enzyme present in Bothrops atrox venom, and 33% identity with that of human thrombin B chain. The most interesting difference between the structures of RVV-V and other trypsin-type serine proteases is that the conservative Ser214-Trp215 Gly216 sequence (chymotrypsinogen numbering), considered as the site of antiparallel beta-sheet formation between the protein substrate and most serine proteases, has been replaced by the corresponding sequence Ala-Gly-Gly. PMID- 3053713 TI - Cloning and sequencing of the yeast gene for dolichol phosphate mannose synthase, an essential protein. AB - Dolichol phosphate mannose (Dol-P-Man) synthase (EC 2.4.1.83) catalyzes the formation of Dol-P-Man from Dol-P and GDP-Man. The structural gene for yeast Dol P-Man synthase (DPM1) was isolated by screening a yeast genomic DNA library for colonies that overexpressed Dol-P-Man synthase activity. This approach relied on a method to screen for Dol-P-Man synthase activity in lysed yeast colonies and used a yeast mutant with very low Dol-P-Man synthase activity in colony lysates. Transformants isolated using this technique expressed Dol-P-Man synthase activity 9-14-fold higher than that of a wild type strain, and all seven plasmids conferring this overproduction had a common region in their yeast genomic DNA insert. DPM1 is the structural gene for yeast Dol-P-Man synthase since Escherichia coli transformants harboring this gene express Dol-P-Man synthase activity in vitro. DNA sequencing of the DPM1 gene revealed an open reading frame of 801 bases. The 30-kDa size of the predicted protein is in excellent agreement with the size of the purified yeast enzyme (Haselbeck, A., and Tanner, W. (1982) Proc. Natl. Acad. Sci. U. S. A. 79, 1520-1524). Analysis of the predicted amino acid sequence reveals the protein has a potential membrane spanning domain of 25 amino acids at its COOH terminus. The protein's NH2 terminus, though not hydrophobic, meets existing criteria for yeast signal sequences, but there is no site for cleavage by signal peptidase. If the NH2 terminus is a functional signal sequence, the protein is predicted to be oriented toward the lumen of the endoplasmic reticulum with both NH2 and COOH termini serving as membrane anchors. If there is no signal sequence, the enzyme is predicted to face the cytoplasm and be anchored only by its COOH terminus. The DPM1 gene is essential for viability in yeast since disruption of the gene is lethal. We suspect Dol-P-Man synthase is not an essential protein due to its role in N-glycosylation since mutations in other genes that affect the late steps in lipid-linked oligosaccharide synthesis do not affect cell growth. Instead, DPM1 may be an essential gene because its product is required for O-glycosylation in yeast or because Dol-P-Man synthase is needed in some unidentified pathway. PMID- 3053714 TI - Expression of an enzymatically active Yb3 glutathione S-transferase in Escherichia coli and identification of its natural form in rat brain. AB - Glutathione S-transferases containing Yb3 subunits are relatively uncommon forms that are expressed in a tissue-specific manner and have not been identified unequivocally or characterized. A cDNA clone containing the entire coding sequence of Yb3 glutathione S-transferase mRNA was incorporated into a pIN-III expression vector used to transform Escherichia coli. A fusion Yb3-protein containing 14 additional amino acid residues at its N terminus was purified to homogeneity. Recombinant Yb3 was enzymatically active with both 1-chloro-2,4 dinitrobenzene and 1,2-dichloro-4-nitrobenzene as substrates but lacked glutathione peroxidase activity. Substrate specificity patterns of recombinant Yb3 were more limited than those of glutathione S-transferase isoenzymes containing Yb1- or Yb2-type subunits. Peptides corresponding to unique amino acid sequences of Yb3 as well as a peptide from a region of homology with Yb1 and Yb2 subunits were synthesized. These synthetic peptides were used to raise antibodies specific to Yb3 and others that cross-reacted with all Yb forms. Immunoblotting was utilized to identify the natural counterpart of recombinant Yb3 among rat glutathione transferases. Brain and testis glutathione S-transferases were rich in Yb3 subunits, but very little was found in liver or kidney. Physical properties, substrate specificities, and binding patterns of the recombinant protein paralleled properties of the natural isoenzyme isolated from brain. PMID- 3053715 TI - Circular dichroism and fluorescence studies on protein synthesis initiation factor eIF-4E and two mutant forms from the yeast Saccharomyces cerevisiae. AB - Circular dichroism studies have shown that eukaryotic initiation factor 4E contains low amounts of alpha-helix; the main elements of secondary structure are beta-sheets/turns and aperiodic regions. Interactions with cap analogs are accompanied by small but reproducible changes in overall secondary structure, which may also involve more significant perturbations of localized regions containing certain phenylalanine residues. Dissociation constants for interactions with nucleotides have been established from fluorescence titrations. Results show that the (N-7) methylated guanosine nucleotides bound more strongly than their nonmethylated counterparts. Involvement of a key tryptophan residue in the cap binding site was suggested. Additional studies with two cap binding mutant forms of the protein, designated SK-4 (W----75----L) and SK-6 (W----115--- L), confirmed and extended these observations. Fluorescence melting experiments indicated that binding of cap analogs stabilized the protein against thermal perturbation and demonstrated subtle differences in folding between the wild-type and mutant forms of the protein. These subtle differences in folding may account for the observed loss in cap specificity of both mutant forms. PMID- 3053716 TI - Comparison of the ligand binding properties of two homologous rat apocellular retinol-binding proteins expressed in Escherichia coli. AB - Cellular retinol-binding protein (CRBP) and cellular retinol-binding protein II (CRBP II) are 132-residue cytosolic proteins which have 56% amino acid sequence identity and bind all-trans-retinol as their endogenous ligand. They belong to a family of cytoplasmic proteins which have evolved to bind distinct hydrophobic ligands. Their patterns of tissue-specific and developmental regulation are distinct. We have compared the ligand binding properties of rat apo-CRBP and apo CRBP II that have been expressed in Escherichia coli. Several observations indicate that the E. coli-derived apoproteins are structurally similar to the native rat proteins: they co-migrate on isoelectric focusing gels; and when complexed with all-trans-retinol, their absorption and excitation/emission spectra are nearly identical to those of the authentic rat holoproteins. Comparative lifetime and acrylamide quenching studies suggest that there are differences in the conformations of apo-CRBP and apo-CRBP II. The interaction of E. coli-derived apo-CRBP and apo-CRBP II with a variety of retinoids was analyzed using spectroscopic techniques. Both apoproteins formed high affinity complexes with all-trans-retinol (K'd approximately 10 nM). In direct binding assays, all trans-retinal bound to both apoproteins (K'd approximately 50 nM for CRBP; K'd approximately 90 nM for CRBP II). However, all-trans-retinal could displace all trans-retinol bound to CRBP II but not to CRBP. These observations suggests that there is a specific yet distinct interaction between these two proteins and all trans-retinal. Apo-CRBP and apo-CRBP II did not demonstrate significant binding to either retinoic acid or methyl retinoate, an uncharged derivative of all-trans retinoic acid. This indicates that the carboxymethyl group of methyl retinoate cannot be sterically accommodated in their binding pockets and that failure to bind retinoic acid probably is not simply due to the negative charge of its C-15 carboxylate group. Finally, neither all-trans-retinol nor retinoic acid bound to E. coli-derived rat intestinal fatty acid-binding protein, a homologous protein whose tertiary structure is known. Together, the data suggest that these three family members have acquired unique functional capabilities. PMID- 3053717 TI - Radical formation in the dimeric small subunit of ribonucleotide reductase requires only one tyrosine 122. AB - The small subunit of ribonucleoside-diphosphate reductase (EC 1.17.4.1) is a homodimer. Its catalytic site contains one tyrosyl radical, which is localized to Tyr-122 in one of its polypeptide chains. The engineered Tyr-122----Phe protein was used to demonstrate that it is possible to form a correct ferric iron center in vitro in the absence of Tyr-122. Heterodimers, consisting of one Tyr-122 containing polypeptide chain and one Phe-122-containing polypeptide chain, were constructed. The heterodimer population contained one-half the amount of tyrosyl radical as compared to a homodimer with Tyr-122, i.e. every second heterodimer contains a tyrosyl radical. Thus, one Tyr-122 is sufficient for radical formation. Radical-containing heterodimers are catalytically competent. PMID- 3053718 TI - Insulin stimulates the translation of ribosomal proteins and the transcription of rDNA in mouse myoblasts. AB - Insulin stimulates the translation of ribosomal protein (r-protein) mRNAs and the transcription of rDNA in mouse MM14DZ myoblasts. Analysis of the distribution of S16, L18, and L32 r-protein mRNAs in polysome gradients indicates that the increased translation of these mRNAs in insulin-treated myoblasts is due to the recruitment of mRNAs that were not previously being translated. In contrast, the translational efficiencies of beta-actin, c-myc, and p31 mRNAs are not affected by insulin. Hybridization analysis of RNA transcribed in nuclear run-on reactions indicates that insulin also stimulates the transcription of rDNA. Both the increases in r-protein translation and rDNA transcription occur coordinately and are maximal within 15 min of insulin treatment of myoblasts. However, insulin has no effect on the rate of cell division or the steady state levels of r-protein mRNAs. Surprisingly, after myoblasts differentiate into fibers, insulin does not affect the r-protein mRNA translation or rDNA transcription. These experiments indicate that the synthesis of the macromolecular components of ribosomes is tightly and coordinately controlled in myoblasts. PMID- 3053719 TI - Conformational changes in the membrane-bound hydrogenase of Bradyrhizobium japonicum. Evidence that the redox state of the enzyme affects its accessibility to protease and membrane-impermeant reagents. AB - The sensitivity of the membrane-bound hydrogenase of Bradyrhizobium japonicum to inactivation by proteases and membrane-impermeant protein modification reagents was compared under hydrogen versus oxygen. In membrane vesicles, the half-life of enzyme inactivation by trypsin of the H2-reduced enzyme was approximately 10 min, whereas O2-oxidized enzyme was much less sensitive to trypsin inactivation (half life of over 90 min). Diazobenzene sulfonate (DABS) affected the enzyme activity in a manner similar to proteases. With DABS, the enzyme had a half-life of 2-3 min under H2 versus over 30 min under O2. Experiments in which the gas phase (containing either H2 or O2) available to the membranes was changed prior to the protease or chemical modification treatments indicated that it is the redox state of the enzyme at the time of the treatment which determines the sensitivity of the enzyme to inactivation. The redox-dependent differences in the behavior of the membrane-bound enzyme were attributed to changes in the accessibility of the small (33 kDa) subunit. The kinetics of enzyme inactivation by trypsin, under H2, correlated very well with the degradation of the intact 33-kDa subunit, whereas the large subunit (65 kDa) was rather resistant to proteolytic degradation. DABS treatment was found to decrease the reactivity of the small subunit to its antibody concomitant with enzyme inactivation under H2, but without such an effect on the O2-oxidized enzyme. In contrast to the results with the membrane bound enzyme, purified dehydrogenase was found to be equally susceptible to inactivation by proteolysis or chemical modification irrespective of whether the treatments were performed under H2 or O2. These results indicate that, in the membrane, hydrogenase undergoes a redox-linked conformational change, whereby the small subunit of the enzyme becomes more accessible to external reagents when the enzyme is in its reduced form. PMID- 3053720 TI - Three-dimensional structure of the tryptophan synthase alpha 2 beta 2 multienzyme complex from Salmonella typhimurium. AB - The three-dimensional structure of the alpha 2 beta 2 complex of tryptophan synthase from Salmonella typhimurium has been determined by x-ray crystallography at 2.5 A resolution. The four polypeptide chains are arranged nearly linearly in an alpha beta beta alpha order forming a complex 150 A long. The overall polypeptide fold of the smaller alpha subunit, which cleaves indole glycerol phosphate, is that of an 8-fold alpha/beta barrel. The alpha subunit active site has been located by difference Fourier analysis of the binding of indole propanol phosphate, a competitive inhibitor of the alpha subunit and a close structural analog of the natural substrate. The larger pyridoxal phosphate-dependent beta subunit contains two domains of nearly equal size, folded into similar helix/sheet/helix structures. The binding site for the coenzyme pyridoxal phosphate lies deep within the interface between the two beta subunit domains. The active sites of neighboring alpha and beta subunits are separated by a distance of about 25 A. A tunnel with a diameter matching that of the intermediate substrate indole connects these active sites. The tunnel is believed to facilitate the diffusion of indole from its point of production in the alpha subunit active site to the site of tryptophan synthesis in the beta active site and thereby prevent its escape to the solvent during catalysis. PMID- 3053721 TI - Load-displacement properties of lower cervical spine motion segments. AB - The load-displacement behavior of 35 fresh adult cervical spine motion segments was measured in compression, shear, flexion, extension, lateral bending and axial torsion tests. Motion segments were tested both intact and with posterior elements removed. Applied forces ranged to 73.6 N in compression and to 39 N in shear, while applied moments ranged to 2.16 Nm. For each mode of loading, principal and coupled motions were measured and stiffnesses were calculated. The effect of disc degeneration on motion segment stiffnesses and the moments required for motion segment failure were also measured. In compression, the stiffnesses of the cervical motion segments were similar to those of thoracic and lumbar motion segments. In other modes of loading, cervical stiffnesses were considerably smaller than thoracic or lumbar stiffnesses. Removal of the posterior elements decreased cervical motion segment stiffnesses by as much as 50%. Degenerated cervical discs were less stiff in compression and stiffer in shear than less degenerated discs, but in bending or axial torsion, no statistically significant differences were evident. Bending moments causing failure were an order of magnitude lower than those for lumbar segments. PMID- 3053722 TI - Analysis of surface damage in retrieved carbon fiber-reinforced and plain polyethylene tibial components from posterior stabilized total knee replacements. AB - The performance of carbon fiber-reinforced ultra-high molecular weight polyethylene was compared with that of plain (non-reinforced) polyethylene on the basis of the damage that was observed on the articulating surfaces of retrieved tibial components of total knee prostheses. Established microscopy techniques for subjectively grading the presence and extent of surface damage and the histological structure of the surrounding tissues were used to evaluate twenty six carbon fiber-reinforced and twenty plain polyethylene components that had been retrieved after an average of twenty-one months of implantation. All of the tibial components were from the same design of total knee replacement. The two groups of patients from whom the components were retrieved did not differ with regard to weight, the length of time that the component had been implanted, the radiographic position and angular alignment of the component, the original diagnosis, or the reason for removal of the component. The amounts and types of damage that were observed did not differ for the two materials. For both materials, the amount of damage was directly related to the length of time that the component had been implanted. The histological appearance of tissues from the area around the component did not differ for the two materials, except for the presence of fragments of carbon fiber in many of the samples from the areas around carbon fiber-reinforced components. PMID- 3053724 TI - Slipped capital femoral epiphysis. PMID- 3053723 TI - Subluxation of the patella. Computed tomography analysis of patellofemoral congruence. AB - Fifty patients who had patellar subluxation and thirty control subjects were examined using axial roentgenograms of the patellofemoral joint that were made with the knee in 30 and 45 degrees of flexion, as well as computed tomography scans that were made with the knee in full extension. The amount of lateral patellar tilt was quantitatively assessed using the lateral patellofemoral angle, as described by Laurin et al., and the congruence angle, as described by Merchant et al. In both the control subjects and the patients, the angle of Laurin et al. changed significantly when the knee was flexed from full extension to 30 degrees. The difference between the groups was statistically significant at each angle of flexion of the knee, and the difference between the groups was most prominent on the computed tomography scans that were made with the knee in full extension (p less than 0.001). In the patients, the average congruence angle (as described by Merchant et al.) was 5 degrees and in the control subjects, -10 degrees. This indicated that, in our patients, the extent of the patellar subluxation was less than that in previously reported series, and, as a result, the sensitivity of the congruence angle in diagnosing patellar subluxation was only 0.30. In contrast, the sensitivity and specificity of the computed tomography scans for diagnosing patellar subluxation were 0.96 and 0.90, respectively--that is, they were higher than the values that were obtained using any axial roentgenograms. Thus, our results indicated that patellar subluxation can be detected more accurately by using computed tomography with the knee in full extension than by using conventional axial roentgenograms. PMID- 3053725 TI - Infection in bone allografts. Incidence, nature, and treatment. PMID- 3053726 TI - On the 85th birthday of this journal. PMID- 3053727 TI - Epidemiology and the prevention of cancer: some recent developments. PMID- 3053728 TI - Review of Ehlers-Danlos syndrome. Successful repair of rupture and dissection of abdominal aorta. AB - Successful surgical treatment of spontaneous rupture and dissection of the abdominal aorta in Ehlers-Danlos syndrome has not been previously reported. A 16 year-old male sustained spontaneous rupture and dissection of the abdominal aorta. Successful surgical treatment included placement of an abdominal aortic bifurcation graft. Genetical, biochemical and clinical differences of seven types of the syndrome are outlined. A brief guideline for treatment and prevention of vascular complications is discussed. PMID- 3053729 TI - A new technique of microlympho-venous anastomoses. Experimental study. AB - This experimental study in dogs presents a new technique of microlympho-venous anastomosis to improve long-term patency rates and clinical results in lymphedema therapy. Technical points, such as an oval window on the wall of the vein and a few sutures piercing only two lymphatic layers, adventitia and media, outside the lumen for successful results are emphasized. Three methods for assessment of patency of anastomoses were used: (1) observation with operating microscope of dye transit across the anastomotic site; (2) lymphography, and (3) histopathologic examination. Of 23 dogs, five were used to perfect the technique, three died after operation, while the remaining 15 were divided into three groups of five dogs each. In the first group which were explored one month after surgery, all anastomoses were patent (100% patency). In the second group, explored three months after surgery, one anastomosis was obstructed and four were patent (80% patency). In the third group, explored six months after surgery, one anastomosis was obstructed and four were patent (80% patency). The results obtained with this technique and the occurrence of a similar physiological lympho venous anastomosis suggest that end-to-side anastomosis may be the technique of choice. PMID- 3053730 TI - Noninvasive assessment of acute rejection after orthotopic heart transplantation: value of changes in cardiac volume and cardiothoracic ratio. AB - Since 1984, 47 orthotopic heart transplantations (HTX) were carried out in 45 patients. In 29 long-term survivors cardiac volume as well as cardiothoracic ratio were measured during their routine follow-up. These two parameters of cardiac size were evaluated from posterior-anterior (pa) and lateral chest x-rays by using conventional technics. Changes of these parameters were correlated to the histological grading of endomyocardial biopsies (EMB). Increases of cardiac volume of more than 10 percent or 100 ml compared with the last measurement and simultaneous increases of cardiothoracic ratio of more than 2 percent were assumed to represent rejection equivalents. Sensitivity and specificity were 0.759 and 0.969, respectively. Predictive values of a positive or negative test for the presence or absence of disease came to 0.815 and 0.957. PMID- 3053732 TI - Transaortic saphenous patch angioplasty for left main coronary artery stenosis. An alternative to coronary artery bypass. AB - We present two cases of isolated proximal left main coronary artery stenosis treated by direct transaortic angioplasty. In selected patients this technique offers a valuable alternative to CAB graft with the advantage of restoring unobstructed antegrade flow. PMID- 3053731 TI - Diagnosis of acute and chronic cardiac rejection by magnetic resonance imaging: a non-invasive in-vivo study. AB - To evaluate the potential usefulness for characterization of tissue and anatomical changes associated with cardiac transplantation rejection by nuclear magnetic resonance imaging (MRI), sixteen dogs underwent heterotropic cardiac transplantation with six not immunosuppressed serving as controls. Myocardial biopsy and MRI were obtained and compared on a weekly basis. Untreated allografts showed a significant increase in T2 and intensity values by MRI compared to the native heart as early as one week after transplantation. The MRI findings corresponded to the histological progression of acute rejection process in both treated and untreated groups. The linear relationship between histology and MRI was 0.72 while the correlation between T2 and the water content was 0.92. Serial gated MRI correlated with chronic anatomical changes of transplant rejection with evidence of progressive or increasing myocardial wall thickness and decrease in ventricular chamber size. PMID- 3053733 TI - Alcohol oxidase expressed under nonmethylotrophic conditions is imported, assembled, and enzymatically active in peroxisomes of Hansenula polymorpha. AB - We have introduced into Hansenula polymorpha an extra copy of its alcohol oxidase gene. This gene which is under the control of the Saccharomyces cerevisiae phosphoglycerate kinase promoter is integrated in a chromosome different from the one containing the endogenous gene. Cells with the extra alcohol oxidase gene, grown on glucose or ethanol as the sole carbon source, express enzymatically active alcohol oxidase. However, other enzymes characteristic for methylotrophic growth conditions are absent or present at low levels. Most of the alcohol oxidase occurs in the octameric state and immuno- and cytochemical evidence shows that it is located in a single enlarged peroxisome per cell. Such peroxisomes show crystalloid inclusions which are lacking in the peroxisomes present in glucose grown control cells. Our results suggest that import into peroxisomes of H. polymorpha, assembly and activation of alcohol oxidase is not conditionally dependent on adaptation to methylotrophic growth conditions and that proliferation of peroxisomes is a well-programmed process that is not triggered solely by overproduction of a peroxisomal protein. PMID- 3053734 TI - Topogenic analysis of the human immunodeficiency virus type 1 envelope glycoprotein, gp160, in microsomal membranes. AB - The orientation in cellular membranes of the 856 amino acid envelope glycoprotein precursor, gp160, of human immunodeficiency virus type 1 was investigated in vitro. Variants of the env gene were transcribed using the bacteriophage SP6 promoter, translated using a rabbit reticulocyte lysate, and translocated into canine pancreatic microsomal membranes. Immunoprecipitation studies of gp160 variants using antibodies specific for various gp160-derived polypeptides provided evidence that the external (cell surface) domain of gp160 begins at the mature amino terminus of the protein and continues through amino acid 665. A stop transfer sequence (transmembrane domain) was identified in a hydrophobic region COOH-terminal to amino acid 665 and NH2-terminal to amino acid 732. Protease protection experiments demonstrated that gp160 possesses a single cytoplasmic domain COOH-terminal to residue 707. Membrane extraction studies using carbonate buffer provided evidence that the 29 amino acid hydrophobic domain (residues 512 541) of gp160 was unable to serve as a stop-transfer sequence. Finally, we propose that the cytoplasmic tail of gp160 forms a secondary association with the microsomal membranes. PMID- 3053735 TI - Exocytosis of vacuolar apical compartment (VAC): a cell-cell contact controlled mechanism for the establishment of the apical plasma membrane domain in epithelial cells. AB - The vacuolar apical compartment (VAC) is an organelle found in Madin-Darby canine kidney (MDCK) cells with incomplete intercellular contacts by incubation in 5 microM Ca++ and in cells without contacts (single cells in subconfluent culture); characteristically, it displays apical biochemical markers and microvilli and excludes basolateral markers (Vega-Salas, D. E., P. J. I. Salas, and E. Rodriguez Boulan. 1987. J. Cell Biol. 104:1249-1259). The apical surface of cells kept under these culture conditions is immature, with reduced numbers of microvilli and decreased levels of an apical biochemical marker (184 kD), which is, however, still highly polarized (Vega-Salas, D. E., P. J. I. Salas, D. Gundersen, and E. Rodriguez-Boulan. 1987. J. Cell Biol. 104:905-916). We describe here the morphological stages of VAC exocytosis which ultimately lead to the establishment of a differentiated apical domain. Addition of 1.8 mM Ca++ to monolayers developed in 5 microM Ca++ causes the rapid (20-40 min) fusion of VACs with the plasma membrane and their accessibility to external antibodies, as demonstrated by immunofluorescence, immunoperoxidase EM, and RIA with antibodies against the 184-kD apical plasma membrane marker. Exocytosis occurs towards areas of cell cell contact in the developing lateral surface where they form intercellular pockets; fusion images are always observed immediately adjacent to the incomplete junctional bands detected by the ZO-1 antibody (Stevenson, B. R., J. D. Siliciano, M. S. Mooseker, and D. A. Goodenough. 1986. J. Cell Biol. 103:755 766). Blocks of newly incorporated VAC microvilli and 184-kD protein progressively move from intercellular ("primitive" lateral) spaces towards the microvilli-poor free cell surface. The definitive lateral domain is sealed behind these blocks by the growing tight junctional fence. These results demonstrate a fundamental role of cell-cell contact-mediated VAC exocytosis in the establishment of epithelial surface polarity. Because isolated stages (intercellular pockets) of the stereotyped sequence of events triggered by the establishment of intercellular contacts in MDCK cells have been reported during normal differentiation of intestine epithelium (Colony, P. C., and M. R. Neutra. 1983. Dev. Biol. 97:349-363), we speculate that the mechanism we describe here plays an important role in the establishment of epithelial cell polarity in vivo. PMID- 3053736 TI - Oligospecificity of the cellular adhesion receptor Mac-1 encompasses an inducible recognition specificity for fibrinogen. AB - Mitogenesis, cellular aggregation, and motility follow upon the interaction of fibrinogen with certain defined cell surface receptors. In addition to circulating platelets and vascular endothelium, monocytes express what appears to be a receptor for fibrinogen. Evidence is presented here that the leukocyte adhesion receptor Mac-1 can be specifically induced to bind fibrinogen with characteristics immunochemically and functionally distinct from the established Arg-Gly-Asp-directed fibrinogen receptors. The competence of Mac-1 as a fibrinogen receptor is a general property of cells of monocyte and myeloid lineage acquired after maturational changes of some regions of the alpha subunit of Mac-1 during the process of cell differentiation. This ligand recognition specificity of Mac-1 is lacking for the resting cell. Rather, induction of fibrinogen binding capacity of Mac-1 is due to a cellular response to selected agonists characterized by inducing rapid transients of cytosolic Ca2+. Although different in activation pathways and recognition specificity, Mac-1 exhibits an oligospecific ligand versatility characteristic of other homologous Arg-Gly-Asp directed adhesion receptors. PMID- 3053738 TI - Video-enhanced light microscopy and its applications in cell biology. AB - The combination of novel optical microscopic techniques with advanced video and digital image-processing technology now permits dramatic improvements in the quality of light-microscope images. Such video-enhanced light microscopy has lead to a renaissance in the applications of the light microscope for the study of living cells in two important areas: the intensification of faint fluorescence images, permitting observation of fluorescently labelled cells under conditions of very low illuminating intensity; and the enhancement of extremely low contrast images generated by minute cellular structures, so that these may be clearly seen and their normal intracellular movements recorded. Application of both these aspects of video-enhanced light microscopy have recently led to major discoveries concerning the functioning of the living cell. In this review I discuss the equipment, procedures and image-processing principles employed in these applications, and describe and illustrate some of the spectacular results that have recently been obtained. PMID- 3053737 TI - Antigenic and functional characterization of a rat central nervous system-derived cell line immortalized by a retroviral vector. AB - We have immortalized rat central nervous system (CNS) cells of primary cultures of rat optic nerve with murine leukemia virus psi-2,SV-40-6, which is defective in assembly and contains the SV-40 large T antigen and neomycin resistance genes, to produce a cell line that we named A7. After drug selection, greater than 90% of the growing cells expressed nuclear SV-40 large T cells and a fraction of these contained the astrocyte-specific marker, glial fibrillary acidic protein. The majority of these cells also expressed surface marker A4 (specific for neural tube derivatives), Ran 2, p185 (the 185-kD phosphoprotein product of the neu oncogene), and fibronectin, but did not express the astrocyte enzymes glutamine synthetase and monoamine oxidase B. Surface markers characteristic of glial progenitors (A2B5) and oligodendrocytes (galactocerebroside) were not detected. After two rounds of cell cloning, subclone A7.6-3 expressed Ran 2, fibronectin, and the neural cell adhesion molecule (N-CAM) but not glial fibrillary acidic protein and A4. The A7 cell line and subclones also displayed certain functions of type 1 astrocytes: the conditioned medium of these cells had a potent mitogenic activity for glial progenitor cells which could be neutralized by anti platelet-derived growth factor antibodies and monolayers of these cells supported the growth of embryonic hypothalamic neurons. We conclude that a retrovirus containing SV-40 large T antigen can immortalize rat CNS cells and that such immortalized glial cells retain at least two important functions of type 1 astrocytes: the ability to secrete platelet-derived growth factor and to support the growth of embryonic CNS neurons. Moreover, such stable immortalized clonal cell lines can be used to study gene regulation in glial cells. PMID- 3053739 TI - MHC control of cell position in vitro. AB - Two-dimensional sorting-out behaviour (segregation) in mixtures of pulmonary endothelial cell lines derived from congenic strains of mice was examined using dense confluent cultures in which mitosis is rare. Cell MHC type was detected by autoradiographic labelling or by immunofluorescence techniques. For autoradiography cultures of one type were previously labelled with [3H]thymidine and one component of the mixed cell types was prepared from these cultures. Autoradiographs were prepared from the fixed cultures. Counts of contiguous neighbours (labelled or unlabelled around a randomly chosen central labelled cell) were made: these were analysed statistically using a new model for such a system. The results show that segregation of a clustering type took place if the alleles of the D locus in the H-2 complex were mismatched in the mixed strains, but that matching at the IC locus (or some locus to the right of IJ) overrides the effect of D mismatch. After 2 days culture sorting-out was easily detectable when the cells were stained for their histocompatibility antigens and groups of up to 12 cells of the same type were associated together. PMID- 3053740 TI - Characterization of a 125K glycoprotein associated with bovine epithelial desmosomes. AB - An analysis of the concanavalinA binding polypeptide components of bovine tongue epithelial desmosomes reveals that in addition to the known desmosomal glycoproteins of 100/115K (the 'desmocollins'), 140K and 160/165K ('desmoglein 1') there is an uncharacterized glycoprotein of 125K (K = Mr x 10(-3). This latter polypeptide is immunologically distinct from known desmosomal glycoproteins, as determined by Western immunoblotting, but is recognized by an antibody preparation directed against the epithelial cell adhesion molecule E cadherin. Moreover, the cadherin antibodies recognize a polypeptide present in bovine muzzle desmosomes that co-migrates with the 125K glycoprotein component of bovine tongue epithelial desmosomes. Upon treatment of bovine tongue desmosomes with a solution containing 9.5 M-urea, the 125K polypeptide becomes enriched in a urea-insoluble, membrane-enriched pelletable desmosomal fraction. Cadherin antibodies and antibodies directed against the 100/115K and 160/165K desmosomal glycoproteins generate similar immunofluorescence staining patterns in cryostat sections of bovine tongue epithelium. However, immunoelectron microscopic analysis of bovine tongue epithelium reveals that cadherin antibodies recognize components located both along the intercellular region of the desmosome and along nondesmosomal cell surfaces whereas antibodies directed against the 100/115K and the 160/165K desmosomal glycoproteins bind specifically to desmosomes. These results suggest that a cadherin-like glycoprotein component may play a role in the adhesive properties of the desmosomes of stratified squamous epithelia. PMID- 3053741 TI - Platelet-activating factor as a mediator of bronchial asthma. PMID- 3053743 TI - Immune markers in the diagnosis of malignant lymphomas. AB - The concept that hematopoietic neoplasms can be related phenotypically and functionally to their normal counterparts in the immune system has greatly improved our understanding of this clinically and morphologically complex group of malignancies. On a practical level, the development of monoclonal antibodies has provided an unlimited supply of highly specific reagents, each reactive with an individual antigenic determinant. Monoclonal antibodies reactive with lymphoid and monocytic subsets can be used in the classification, primary diagnosis and staging of hematopoietic malignancies. PMID- 3053742 TI - An improved membrane-filtration enzyme immunoassay for the rapid serological diagnosis of viral infections. AB - A one-step modification of the membrane-filtration enzyme immunoassay (MF EIA) (Barnett et al., J. Clin. Microbiol., 23:385-399, 1987), for estimation of virus specific antibody is described. The modified MF EIA allowed serum, antigen and enzyme-conjugated anti-globulin to be incubated together in membrane-based 96 well plates to enable the formation of immune complexes in solution at 37 degrees C. The assay required only 45 min for completion and polyethylene glycol was shown to be an essential component in reaction mixtures for IgG assays to enhance immune complex formation. The modified MF EIA was as sensitive as the previous two-step method for monitoring responses to influenza vaccine, and control antigen backgrounds were significantly reduced. The one-step procedure was also shown to be suitable for the rapid serodiagnosis of naturally acquired influenza A and B infections. However, MF EIA detected cross-reactive H1N1 responses in 57.7% of naturally-acquired H3N2 infections, suggesting that responses to common internal antigens were being measured. Cross-reactive responses to influenza A viruses could not be detected in volunteers receiving subunit vaccines. PMID- 3053744 TI - The molecular analysis of chromosomal translocations as a diagnostic, epidemiological and potentially prognostic tool in lymphoid neoplasia. AB - Neoplasms have been shown to be associated with specific non-random chromosomal abnormalities. The present paper summarizes molecular consequences of chromosomal translocations in lymphoid malignancies, especially in Burkitt's lymphoma. Among such consequences are derangements of the regulation of oncogene transcription. Besides their possible importance for lymphoma pathogenesis, chromosomal translocations are associated in some lymphoid neoplasms with a worse prognosis. PMID- 3053745 TI - Human herpesvirus-6 (HHV-6, HBLV) in sarcoidosis and lymphoproliferative disorders. AB - Serologic studies were done to estimate the antibody prevalence against human herpesvirus 6 (HHV-6) in patients with malignant lymphomas, Sjogren's syndrome and sarcoidosis. Serologic studies showed IgG antibody titers against HHV-6 in up to 41% of patients with sarcoidosis, 50-70% with malignant lymphomas and in 36% with Sjogren's syndrome. In situ hybridization on lymph node biopsies was positive for HHV-6 genome in 1 out of 5 sarcoidosis lymph nodes. PMID- 3053746 TI - Chronic herpes simplex virus and varicella zoster virus infection. AB - After defining such terms as persistent and chronic infection, latency, recurrence, recrudescence, and exogenous reinfection they are applied to infections with HSV and VZV. Possible factors determining pathogenicity are discussed, and an overview is given of the wide range of illnesses and case reports ascribed to HSV and VZV infections. Various types of infection afford different diagnostic procedures. Besides virus isolation supplemented by viral antigen identification IgG antibody tests (increase in titer) may be useful. IgG subtype and IgA antibody determinations appear to be of limited value. Despite the rather large number of available tests, there are still considerable shortcomings in their ultimate significance as to the patient's disease. Thus, some new experimental approaches are mentioned. PMID- 3053747 TI - Advantageous metabolic effects of pulsatile insulin delivery in noninsulin dependent diabetic patients. AB - This study was done to compare the actions of pulsatile and continuous insulin administration in eight noninsulin-dependent diabetic patients. Human insulin was delivered in a pulsatile manner (1.3 mU/kg.min for 2 min, followed by 11 min during which no insulin was infused) or continuously (0.2 mU/kg.min) for 325 min. Endogenous hormone secretion was inhibited by somatostatin (125 micrograms/h), and glucagon was replaced at rate of 3.5 micrograms/h. Under these conditions plasma C-peptide levels fell progressively to extremely low values at the end of the experiment. Continuous insulin infusion resulted in steady plasma insulin levels, averaging 86 pmol/L, while during intermittent insulin administration plasma insulin levels were 5.7 and 158 pmol/L before and 3 min after the start of the insulin injection, respectively. Basal plasma glucagon [mean 158 +/- 11 (+/- SE) vs. 163 +/- 21 ng/L; P = NS] levels were similar on both occasions. During replacement peripheral plasma glucagon levels were no different whatever the mode of insulin administration, nor did they differ from the basal values. The mean plasma glucose concentrations were similar before both studies and rose to 9.5 and 8.6 mmol/L in the first 65 min during continuous and pulsatile insulin administration, respectively. In contrast, during the last 65 min, plasma glucose averaged 6.2 mmol/L during both studies. The glucose infusion rate initially increased, but then rapidly fell to values close to zero at the end of the first 65 min during the continuous insulin infusion, whereas during this time it averaged 0.59 +/- 0.10 mg/kg.min (32.5 +/- 5.5 mumol/kg.min) during pulsatile insulin administration. In the last 65 min the glucose infusion rate was significantly higher during pulsatile than during continuous insulin delivery. Furthermore, pulsatile rather than continuous insulin administration significantly reduced plasma triglyceride, very low density lipoprotein triglyceride, and FFA levels and increased high density lipoprotein cholesterol and apolipoprotein-B levels. We conclude that pulsatile insulin delivery has advantageous metabolic effects compared to continuous hormone administration in patients with noninsulin-dependent diabetes mellitus. PMID- 3053748 TI - Peripheral insulin parallels changes in insulin secretion more closely than C peptide after bolus intravenous glucose administration. AB - The changes in peripheral serum insulin and plasma C-peptide levels and in the insulin secretory rate in response to iv glucose (0.5 g/kg BW) administration were studied in seven normal subjects. Insulin secretory rates were calculated according to a two-compartment model of distribution for C-peptide, using individual C-peptide kinetics calculated from iv bolus injections of biosynthetic human C-peptide. The mean plasma glucose level increased from a fasting level of 5.1 +/- 0.1 (+/- SE) to a peak of 24.0 +/- 1.0 mmol/L at 3 min and reached basal levels 101 +/- 6 min after glucose administration. The mean serum insulin value increased from 50 +/- 12 to a peak of 405 +/- 58 pmol/L at 3 min and then declined to fasting levels 139 +/- 14 min after the stimulus. In contrast, the mean plasma C-peptide level increased from 390 +/- 50 to a peak of 1460 +/- 210 pmol/L at 3 min and only began declining 45 min after glucose administration, reaching fasting levels 191 +/- 15 min after the stimulus. The mean insulin secretory rate increased from 69.8 +/- 19.9 to a peak of 1412.7 +/- 159.1 pmol/min at 3 min (15.3 +/- 2.5-fold elevation over baseline) and reached basal levels 135 +/- 12 min after the stimulus. The clearance of endogenous insulin during the basal period (2.505 +/- 0.365 L/min) and that during the 4 h after the stimulus (2.319 +/- 0.230 L/min) were similar. In conclusion, after bolus iv glucose administration: 1) the insulin secretory rate is more closely represented by changes in peripheral serum insulin than in plasma C-peptide levels; and 2) no change in endogenous insulin clearance occurs. PMID- 3053749 TI - Quantitation of forearm glucose and free fatty acid (FFA) disposal in normal subjects and type II diabetic patients: evidence against an essential role for FFA in the pathogenesis of insulin resistance. AB - This study was designed to quantitate glucose and FFA disposal by muscle tissue in patients with type II diabetes and to investigate the relationship between FFA metabolism and insulin resistance. The forearm perfusion technique was used in six normal subjects and two groups of normal weight diabetic patients, i.e. untreated (n = 8) and insulin-treated (n = 6). The latter received 2 weeks of intensive insulin therapy before the study. Plasma insulin levels were raised acutely [950-1110 pmol/L) (130-150 microU/mL)], while the blood glucose concentration was clamped at its basal value [4.9 +/- 0.1 (+/- SE) mmol/L in the normal subjects, 5.7 +/- 0.5 in the insulin-treated diabetic patients, and 5.5 +/ 0.3 in the untreated diabetic patients] by a variable glucose infusion. During the control period, arterial FFA concentrations were similar in the three groups, and they decreased to a comparable extent (less than 0.1 mmol/L) in response to insulin infusion. During the control period, the mean forearm FFA uptake was 2.5 +/- 0.5 mumol/L.min in the normal subjects, 2.9 +/- 0.5 in the insulin-treated patients, and 2.1 +/- 0.5 in the untreated diabetic patients. During the insulin infusion, FFA uptake was profoundly suppressed to similar levels in the normal subjects (0.9 +/- 0.1 mumol/L.min), the insulin-treated diabetic patients (1.1 +/ 0.3), and the untreated diabetic patients (0.9 +/- 0.1; P less than 0.001). Forearm glucose uptake was similar in the three groups during the control period. It increased during the insulin infusion, but the response in both diabetic groups was less than that in the normal subjects. The total amounts of glucose taken up by the forearm during the study period were 5.2 +/- 0.7, 2.6 +/- 0.5, and 2.1 +/- 0.6 mmol/L.min in the normal subjects, the insulin-treated diabetic patients, and the untreated diabetic patients, respectively (P less than 0.01). We conclude that 1) insulin-mediated glucose uptake by forearm skeletal muscle is markedly impaired in type II diabetes and improves only marginally after 2 weeks of intensive insulin therapy; 2) in contrast, no appreciable abnormality in forearm FFA metabolism is demonstrable in insulin-treated type II diabetic patients; and 3) FFA do not contribute to the insulin-treated skeletal muscle insulin resistance that occurs in patients with type II diabetes mellitus. PMID- 3053750 TI - Insulin resistance and beta-cell dysfunction in aging: the importance of dietary carbohydrate. AB - Aging is associated with a progressive decrease in glucose tolerance. This decrease is associated with insulin resistance and beta-cell dysfunction. This study was performed to evaluate the possible role of dietary factors in the glucose intolerance of aging. Two groups of men were studied: one young (Y; n = 8; age range, 18-36 yr) and one elderly (E; n = 10; age range, 65-82 yr). Frequently sampled iv glucose tolerance tests were performed in random order: 1) during ad libitum home dietary conditions; 2) after a 3- to 5-day regimen of very high (85%) carbohydrate intake; and 3) after a 3- to 5-day regimen of low (30%) carbohydrate intake (Y only). From the frequently sampled iv glucose tolerance test data, we calculated the glucose disappearance rate (Kg) and metabolic parameters according to the minimal model method, including the insulin sensitivity index (S1) and the first and second phase beta-cell responsivity to glucose (phi 1 and phi 2). The elderly men, while eating an ad libitum diet, were less tolerant to glucose than the young [mean Kg: E = 1.5 +/- 0.2% (+/- SE) min 1; Y = 2.3 +/- 0.3% min-1; P less than 0.025], had relative insulin resistance (mean Si: Y = 6.1 +/- 1.1; E = 2.4 +/- 0.7 min-1 10(-4)/(microU/mL) [0.85 +/- 0.15 vs. 0.33 +/- 0.10 min-1 10(-4)/(pmol/L)]; P less than 0.01), and lesser second phase beta-cell responsiveness to glucose (mean phi 2: Y = 18.5 +/- 3.6; E = 8.7 +/- 2.7 (microU/mL).min-2/(mg/dL) [2390 +/- 465 vs. 1120 +/- 349 (pmol/L).min-2/(mmol/L)]; P less than 0.05). A maximum improvement in Kg and S1 occurred at 41% carbohydrate feeding in the young men, whereas in the elderly men there was a significant increase in both of these parameters while eating the very high (85%) carbohydrate diet. Thus, the difference in glucose tolerance between groups was corrected by the very high carbohydrate diet (mean Kg: Y = 2.2 +/- 0.2%; E = 2.0 +/- 0.3%/min; P greater than 0.05), as was the age-related difference in insulin sensitivity (mean S1: Y = 5.6 +/- 1.2; E = 4.4 +/- 1.3 min 1 10(-4)/(microU/mL) [0.78 +/- 0.17 vs. 0.61 +/- 0.18 min-1 10(-4)/(pmol/L)]; P greater than 0.5).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3053751 TI - Thyroid volume and serum thyroglobulin levels in patients with acromegaly: correlation with plasma insulin-like growth factor I levels. AB - The pathogenesis of the goiter that is frequently found in patients with acromegaly is not known. Using ultrasonic scanning, we measured thyroid volume in 17 euthyroid patients with acromegaly and examined the relationships among thyroid size, plasma GH and insulin-like growth factor (IGF-I) levels, and serum thyroglobulin (TG) levels. The mean estimated thyroid volume in these 17 patients was 32.8 +/- 15.5 (+/- SD) mL, significantly larger than that in normal subjects (15.4 +/- 3.1 mL), and 64.7% of the patients had multinodular goiter, as identified by ultrasonography. Thyroid volume was positively correlated with plasma GH and IGF-I levels and heel-pad thickness, but not with the serum TSH level. In 7 patients, thyroid volume decreased in association with a decline in plasma GH and IGF-I levels after surgical treatment. The serum TG level was elevated in 7 of the 15 patients in whom it was measured, and the mean value was 51.7 +/- 62.7 (+/- SD) micrograms/L (normal, 12.6 +/- 6.4 micrograms/L). We found no correlations among the serum TG and TSH levels, plasma GH and IGF-I concentrations, and/or thyroid volume. However, serum TG decreased after surgical treatment, just as did plasma IGF-I. These observations together with the results of recent in vitro studies by others suggest that IGF-I is one of the factors involved in goiter formation, but the elevated serum TG levels in acromegaly are controlled not only by IGF-I but also by other factors. PMID- 3053752 TI - Resistance of tumor cells to tumor necrosis factor. AB - TNF-alpha is clearly an important mediator of in vitro tumor cell cytotoxicity induced by the activated macrophage. There are a number of other nonspecific mediators of tumor cell cytotoxicity. These include natural killer cells (51, 52), lymphokine-activated killer cells (53), and natural cytotoxic cells (54). The role that TNF-alpha may play in the cytotoxicity induced by these cell types has not been completely elucidated. Neither is it known what role, if any, TNF alpha may play in major histocompatibility-restricted (T cell)-mediated tumor cell cytotoxicity. Just as in the case of the activated macrophage, activated cytotoxic T cells produce a number of mediators that inhibit the growth of tumor cells or that induce tumor cell cytotoxicity (55). The role that TNF-alpha plays in the whole process of the regulation of tumorigenesis will not become completely defined until an appropriate set of genetic experiments is completed which utilizes transplantable tumor cell lines selected specifically for resistance to this cytokine in in vivo tumor models. The prominance of TNF-alpha as a mediator of macrophage-induced tumor cell cytotoxicity makes it a candidate for analysis in studies of the early stages of tumorigenesis. We have chosen to study mechanisms of resistance to this monokine. Our results have shown that there are multiple pathways leading to resistance to TNF-alpha-induced tumor cell cytotoxicity. These pathways include the production of transforming growth factors by tumor cells and the amplified expression of certain oncogenes. Other pathways will undoubtedly become elucidated as we begin to define the molecular mechanisms giving rise to the resistant phenotype. PMID- 3053753 TI - IgM antibody responses to the circumsporozoite protein in naturally acquired falciparum malaria. AB - The circumsporozoite (CS) protein on the surface of sporozoites is the major target for antibody response(s) to the infective stage of the malaria parasite. Sera from malaria endemic areas contain both IgM and IgG antibodies that react with a dominant epitope in the tetrapeptide repeat region of the CS protein. In order to characterize the IgM CS antibody response in Plasmodium falciparum (PF) malaria, a prospective study was conducted in Thai Rangers. Variable IgM responses against the CS protein were detected in 81% of 47 PF-infected subjects. Similar to IgG response kinetics, IgM levels rose to a peak 6-10 days after detection of parasitemia and showed an apparent serum half-life of less than 25 days. The classic difference in isotype ratio (IgG/IgM) between primary and secondary antibody responses was observed to blood-stage, but not CS, antigens. PMID- 3053754 TI - Pre- and postnatal findings in Pena Shokeir I syndrome: case report and a review of the literature. AB - The Pena Shokeir type I syndrome is considered to be a lethal disorder in most cases. Infrequently, some of the affected children may reach the age of 1 year and beyond. When there is a history of another affected sib, the entity can be suspected prenatally. It is an uncommon autosomal recessive disorder. Out of 33 reported infants reviewed by us, six survived the neonatal period; among a further 27 cases, reported more recently, none survived the neonatal period. The clinical findings at birth are multiple and involve mainly the musculoskeletal and respiratory systems, accompanied by characteristic facial changes. The possibility of a primary hereditary malformation affecting the motor neuron cells of the spinal cord is postulated with the subsequent changes representing a fetal akinesia deformation sequence. PMID- 3053755 TI - Pulsatile gonadotrophin releasing hormone therapy in patients with hyperandrogenaemia or hypothalamic amenorrhoea. AB - From 1984 to 1985, 18 patients with infertility and oligomenorrhoea were treated with pulsatile GnRH administration (Zyklomat). According to the hormone levels and the ultrasonographic observation of the ovaries, they could be divided into two categories, group A (n = 11), patients with hyperandrogenaemia, and group B (n = 7), patients with hypothalamic amenorrhoea. As in hyperandrogenaemic patients a pathological LH-secretion pattern was suspected, assessment of LH pulsing (5 ml blood samples at 10 min intervals over 6 h) was performed in this group of patients followed by an oestrogen-gestagen (E-Ge) suppression. One day before discontinuation of this medication, the GnRH pump was applied intravenously. Ovulation induction was more successful in group B than in group A. Hyperandrogenaemic women, in whom ovulation could be induced by the GnRH pump, exhibited higher basal concentrations of FSH, LH, LH/FSH ratio, oestradiol- 17 beta and testosterone (T) than the women not responding to pulsatile GnRH administration. The suppression of T and LH/FSH ratio with E-Ge treatment was more pronounced, while the non-responders had higher basal prolactin concentrations as well as after E-Ge therapy and a significantly greater body weight. The results indicate that GnRH therapy in hypothalamic amenorrhoea is more successful than in hyperandrogenaemia. Overweight hyperandrogenaemic patients appeared to be unsuitable for GnRH treatment, even after previous suppression of the hypothalamic pituitary ovarian axis with E-Ge. PMID- 3053757 TI - Enzyme-linked immunosorbent assay for diagnosis of chronic Q fever. AB - From 1982 through 1987 we diagnosed 13 chronic Q fever cases. Clinically these patients presented a culture-negative endocarditis, and all but two had high complement-fixing antibody titers to Coxiella burnetii phase I (reciprocal titer above 200). With the enzyme-linked immunosorbent assay (ELISA), titers of immunoglobulin G (IgG) to phases I and II of C. burnetii averaged 158,000 and 69,900, respectively, whereas they reached 300 and 3,200 in acute Q fever cases. Similarly, IgA to both phases of C. burnetii and IgM to phase I were consistently higher during chronic than acute Q fever. The serological follow-up of one patient with chronic Q fever over a 4-year period showed a good correlation between the titers of IgG and IgM antibody titers detected by ELISA and indirect fluorescent-antibody test (IFA) to both phases of C. burnetii. Few discrepancies appeared with IgA. Shortly after initiation of antibiotic treatment, a slow and steady decrease of the antibody titers to C. burnetii phases I and II was observed. The complement fixation, IFA, and ELISA tests showed the same type of antibody response. The ELISA proved to be an excellent diagnostic test for chronic Q fever. It distinguished negative from positive reactions clearly, and results were highly reproducible. The reading is objective, and the test is simple to perform and more sensitive than the IFA and complement fixation tests. The ELISA is recommended for serologic evaluation of patients with chronic Q fever. PMID- 3053756 TI - Biotyping coagulase-negative staphylococci. AB - The biochemical profiles obtained with Staph-Ident (Analytab Products, Plainview, N.Y.) panels were combined with the results of adherence and synergistic hemolysis tests to define biotypes among 1,064 clinical isolates representing eight species of coagulase-negative staphylococci. The 672 isolates of Staphylococcus epidermidis were aligned in 69 of 144 potential biotypes in our scheme because of 18 different biochemical profiles and the eight physiologic subtypes. Isolates of most other species were in fewer biotypes because of more uniform adherence and synergistic hemolysis data, as well as fewer biochemical profiles. Since adherence and synergistic hemolysis may prove to be related to virulence and pathogenicity, biotyping with these test results would help evaluate the reliability of adherence and synergistic hemolysis as possible indices of the clinical significance of some of these organisms. When the antimicrobial susceptibility and plasmid profiles obtained on two clusters of S. epidermidis isolates were compared with the biotyping results, one cluster was not further differentiated by plasmid profiles, but was by antimicrobial profiles; the other cluster with only two biotypes was further divided into five distinct types by plasmid profiles but was not separated at all by antimicrobial profiles. PMID- 3053759 TI - Microtechnique for serum opacity factor characterization of group A streptococci adaptable to the use of human sera. AB - We have developed a microtechnique for detection of streptococcal serum opacity factor (OF) and for typing of group A streptococci by inhibition of OF. This technique, which involves the use of single wells of standard 96-well tissue culture plates, offers several advantages over previous methods: no advance test preparation is required, allowing tests to be quickly and easily performed; only small quantities of reagents are required; results can be determined visually (qualitative) or by using a photometric enzyme-linked immunosorbent assay plate reader (quantitative); and human serum samples may be quickly and easily screened for OF-inhibitory antibody and subsequently used in place of difficult-to-produce and expensive hyperimmune animal sera for OF characterization of group A streptococci. Fifty-eight samples of normal adult human serum were tested by this new microtechnique for anti-OF antibodies, and 49 (84%) were found to have antibody against 1 or more of the 27 recognized OF-positive serotypes. OF antibodies to M-4, M-2, M-75, and M-48 were most common in these individuals. These 58 human serum samples collectively contained antibody to 25 of the 27 different OF-producing serotypes. Serum samples from four individuals were tested for persistence of OF antibody. OF antibodies to eight different serotypes present in the serum samples collected 7 to 12 years previously were present in the freshly collected sera, indicating that OF antibody persists in human antisera for many years. This new technique has distinct advantages and makes it possible for many laboratories to use this technique to characterize group A streptococci. PMID- 3053760 TI - Evaluation of different fixatives and treatments for immunohistochemical demonstration of Coxiella burnetti in paraffin-embedded tissues. AB - Various fixatives and treatments such as acetone, methanol, Bouin fixative, modified Bouin fixative, 10% Formalin, modified methacarn, periodate-lysine paraformaldehyde, acetone-methyl benzoate-xylene, and EDTA were evaluated for their effect on the immunoreactivity of Coxiella burnetii in paraffin-embedded tissues by using the avidin-biotin-peroxidase complex and the peroxidase antiperoxidase procedure. C. burnetii antigen was shown to be present in liver, spleen, and uterus tissues of experimentally infected mice by all methods of fixation and treatment. A positive immunoreaction was seen in cytoplasmic vacuoles of macrophages, as extracellular rod-shaped organisms, and as residual particulate extra- and intracellular debris. Immunoreactivity and cellular preservation, however, varied substantially with the individual fixatives. Optimal immunostaining of C. burnetii was achieved by EDTA treatment and Bouin and acetone fixation. The avidin-biotin-peroxidase technique proved to be slightly more sensitive than the peroxidase-antiperoxidase procedure when primary antibody dilution was used as the criterion for sensitivity. PMID- 3053758 TI - Characterization of Escherichia coli serotype O157:H7. AB - A total of 174 strains of Escherichia coli serotype O157:H7 representing human isolates obtained from outbreaks and sporadic cases of hemorrhagic colitis, hemolytic-uremic syndrome, and nonbloody diarrheal illnesses as well as from asymptomatic carriers across Canada and the United States were examined. E. coli serotype O157:H7 possessed distinct biochemical markers, a 100% negative reaction for beta-glucuronidase and sorbitol, and a 100% positive reaction for raffinose and dulcitol; all strains otherwise were biochemically typical of E. coli. The vast majority (97%) of the strains were susceptible to commonly used antimicrobial agents. All strains produced readily detectable levels of Verotoxin; however, with polymyxin extraction, nearly 50% of the strains showed up to a 10-fold increase in the toxin level. None were found to mediate hemagglutination of human group A erythrocytes with or without D-mannose. The majority (approximately 70%) of the strains showed localized and diffuse adherence to HEp-2 cells and Henle 407 cells, and the adherence patterns were not very different from those observed among other E. coli strains. Twenty phage types were recognized, with phage types 1 and 2 accounting for 65% of the test strains. Plasmid analysis indicated three basic plasmid profiles: profile I was characterized by 68.7- and 4.2-megadalton (MDa) plasmids (62% of strains), profile II was characterized by 66.2- and 1.8-MDa plasmids (20% of strains), and profile III was characterized by a 62.5-MDa plasmid (18% of strains). A small number (19%) of the strains carried at least one additional plasmid over the basic complements, and these could be considered to constitute a miscellaneous category. None of the above-described characteristics of E. coli serotype O157:H7 could be directly correlated with one another, with the nature of infection, or with the geographical distribution of strains. PMID- 3053762 TI - Plasmid analysis of Shigella dysenteriae type 1 isolates obtained from widely scattered geographical locations. AB - Plasmid profiles and antimicrobial susceptibility patterns of 343 strains of Shigella dysenteriae type 1, obtained from 18 different geographical locations, were analyzed. Three plasmids, with molecular sizes of 140, 6, and 2 megadaltons (MDa), were present in 94, 98, and 96%, respectively, of the 343 strains isolated during either epidemic or nonepidemic periods from 1965 to 1987. In addition to these plasmids, 83% of the strains harbored a 4-MDa plasmid and 25% harbored a 20 MDa plasmid. Various plasmid profiles were observed in which the 140-, 6-, and 2 MDa plasmids occurred commonly, irrespective of the place of isolation and drug resistance pattern of the strains. Certain profiles showed significant association with drug resistance patterns. These findings suggest that three plasmids, of molecular sizes 140, 6, and 2 MDa, are unique to S. dysenteriae type 1 strains and may indicate the global spread of a pathogenic bacterial clone. Additionally, these core plasmids, plus plasmids of various other sizes, could be used to identify emerging subclones which are causing both epidemic and sporadic disease. Thus, plasmid profiles of S. dysenteriae type 1 strains can be used to monitor possible pandemic strains as well as individual epidemic strains. PMID- 3053761 TI - Determination of serum bactericidal activity against Escherichia coli by an automated photometric method. AB - The resistance of gram-negative bacteria to complement-mediated serum activity is supposedly an important virulence factor. However, the lack of standardization in the methods used to determine serum activity and the many definitions applied make the comparisons between studies very difficult. We developed a rapid photometric method that we compared with a classical killing one. Escherichia coli in the exponential phase of growth in brain heart infusion broth (final inoculum, 10(7) CFU/ml) at 35 degrees C was added to 50% human serum in Veronal buffer. Viable counts and automatic recording of the variations in the optical densities were obtained for 40 E. coli strains isolated from the stools of healthy adults. With the viable count method, 17 (42.5%) were susceptible (at least a 1 log CFU/ml decrease), 17 (42.5%) were resistant (a 0.6 log CFU/ml increase), 4 (10%) were intermediate (poorly growing inoculum or a decrease of less than 1 log CFU/ml), and 2 could not be classified (nonreproducible results). Agreement between both methods was observed for 87.5% of the stool strains. Eight reference strains of known susceptibilities were classified identically by both methods, leading to a final concordance rate of 89.6%. A total of 129 blood isolates were tested by the photometric method: 64 (49.6%) were resistant, 50 (38.8%) were susceptible 5 (3.9%) showed early regrowth, and 10 (7.7%) were not perfectly reproducible. Of these 129 blood isolates, 5 were also tested by the killing method: 37 (49%) were resistant, 32 (43%) were susceptible, and 6 (8%) were intermediate. The concordance rate between both assays was 89% for the blood isolates; when the minor discordances were ruled out, it was 97%. This automated method could be a useful screening tool for detecting resistance to serum in clinical trials and for studying the in vitro variations of this property. PMID- 3053763 TI - Immunoblot analysis of serologic response to outer membrane proteins of Escherichia coli in elderly individuals with urinary tract infections. AB - We used immunoblotting to examine the serologic antibody responses to outer membrane proteins (OMP) of Escherichia coli in both symptomatic and asymptomatic elderly subjects with urinary tract infections. Controls with no present or past urinary tract infections showed variable weak immunoglobulin G (IgG) antibodies to OMP of infecting strains. Elderly individuals with asymptomatic infections demonstrated antibody to both lipopolysaccharide (LPS) and OMP of their infecting strain, with consistent cross-reactivity to OMP of other infecting strains. Young females with acute pyelonephritis showed an IgG response to LPS and OMP with cross-reactivity to OMP of other strains. Elderly individuals with symptomatic invasive infections had strong reactions to both LPS and OMP in specimens collected during the acute phase, generally with an increase in intensity in specimens from convalescent patients. They also demonstrated extensive cross reactivity to LPS and OMP from all other infecting strains. IgM antibody was not observed in any patients. These data confirm other reports of low levels of antibodies to OMP of E. coli in normal populations. Asymptomatic bacteriuria in this population is associated with antibody responses to the LPS and OMP of the infecting strain. Elderly individuals with invasive infections had initial reactions to the infecting strain with an apparent increase in intensity during convalescence. Antibodies to the major OMP appear to be broadly cross-reactive. PMID- 3053764 TI - Isolation and characterization of monoclonal antibodies to Shiga-like toxin II of enterohemorrhagic Escherichia coli and use of the monoclonal antibodies in a colony enzyme-linked immunosorbent assay. AB - The major obstacle in large-scale epidemiological investigations of the incidence of Shiga-like toxin (SLT)-producing Escherichia coli in diarrheal stools is the lack of a rapid, specific test to detect toxin. Enterohemorrhagic E. coli produces elevated levels of SLT-I, SLT-II, or both cytotoxins (also called Verotoxins). SLT-I but not SLT-II can be neutralized by antiserum to purified Shiga toxin and by monoclonal antibodies to the B subunit of SLT-I. In this study, monoclonal antibodies were generated against a crude preparation of SLT-II produced by an E. coli K-12 strain lysogenized with the 933W toxin-converting phage of enterohemorrhagic E. coli 933. Hybridoma culture supernatants were screened for anti-SLT-II antibodies by a cytotoxicity neutralization assay and by an enzyme-linked immunosorbent assay (ELISA). Of 53 ELISA-positive lines, 5 were capable of neutralizing the cytotoxicity of SLT-II but not of SLT-I, Shiga toxin, or a variant of SLT-II produced by E. coli that causes edema disease of swine. All five monoclonal antibodies immunoprecipitated the isolated A subunit of SLT II but not the B subunit. Of these five neutralizing monoclonal antibodies, four were of the immunoglobulin M class and one belonged to the immunoglobulin G1 subclass. All five lines had kappa light chains. These neutralizing monoclonal antibodies have been used as probes in a colony ELISA to detect SLT-II-positive bacterial colonies. The colony ELISA with these monoclonal antibodies is a specific, sensitive test with potential diagnostic value. PMID- 3053765 TI - Possible common biological and immunological properties for detecting encapsulated strains of Staphylococcus epidermidis. AB - Twenty strains of capsular type II Staphylococcus epidermidis, determined by the method of Ichiman, were obtained from clinical specimens. Among them, 5, 5, and 10 strains were 4+, 3+, and 2+ in the intensities of their reactions against fluorescent antibody, respectively. Strains exhibiting 4+ and 3+ intensities were mouse virulent and phage nontypable, while 2+ strains were mouse avirulent and phage typable. When three strains randomly selected from each of the mouse virulent and mouse-avirulent strains were compared in terms of their cell volume indices, all mouse-virulent strains had significantly higher indices (average, 1.86 times) than the mouse-avirulent strains. With intraperitoneal injection of the strains into mice, strains with higher cell volume indices resisted ingestion by peritoneal cells, while strains with low cell volume indices were sensitive to phagocytosis. When the capacity to absorb a definite amount of passive protective activity in rabbit antiserum prepared with capsular type II strains was compared among these strains, 10 to 20 mg of mouse-virulent strains was capable of completely absorbing the passive protective activity, whereas more than 80 mg of the cells was required for similar absorption by mouse-avirulent strains. In ultra-thin sections of three mouse-virulent strains stained with ferritin conjugated rabbit antiserum, well-defined capsules were detected around cell walls; however, no capsule was seen around the walls of three mouse-avirulent strains. PMID- 3053766 TI - Identification of enterotoxigenic Escherichia coli isolates with enzyme-labeled synthetic oligonucleotide probes. AB - Commercially available kits containing alkaline phosphatase-labeled oligonucleotide probes for Escherichia coli heat-stable enterotoxins (STI-H, STI P, and STII) and the heat-labile enterotoxin were compared with bioassays and radiolabeled recombinant DNA probes to identify enterotoxigenic E. coli from 100 clinical isolates. There was very good agreement between the three methods. PMID- 3053768 TI - Disseminated infection caused by urease-negative Cryptococcus neoformans. AB - We report a case of fungemia and disseminated disease caused by a urease-negative strain of Cryptococcus neoformans in a patient with the acquired immune deficiency syndrome. Except for failure to hydrolyze urea, the microbiological characteristics of the isolate were typical of C. neoformans. Laboratory specialists should be aware of the occurrence of atypical strains of C. neoformans, particularly those recovered from patients with the acquired immune deficiency syndrome. PMID- 3053769 TI - Improved detection of heat-labile enterotoxin of enterotoxigenic Escherichia coli by using a commercial coagglutination test. AB - Escherichia coli strains grown on lincomycin-supplemented Mundell agar and on blood agar were compared for their ability to produce heat-labile enterotoxin, as detected by a commercial coagglutination kit. The special agar allowed more strains to be detected, and the results were much more clear-cut. PMID- 3053767 TI - Lack of Shiga-like cytotoxin production by enteroinvasive Escherichia coli. AB - Enteroinvasive Escherichia coli has not been extensively studied for cytotoxin production. We evaluated 30 well-characterized enteroinvasive E. coli strains of all the known invasive serogroups from several geographic regions for their ability to produce Shiga-like cytotoxic activity assayed in a HeLa cell system. None of these strains produced cytotoxic activity that was neutralizable with antibody to Shiga-like toxin I or II. PMID- 3053770 TI - Mannose-resistant hemagglutination of human erythrocytes by enterotoxigenic Escherichia coli with colonization factor antigen II. AB - The human erythrocyte receptor which mediates mannose-resistant hemagglutination by enterotoxigenic Escherichia coli possessing colonization factor antigen II is not universally distributed among donors. Mannose-resistant hemagglutination positive erythrocytes are more common among black donors than nonblack donors; tests with erythrocytes of known antigenic makeup confirm this correlation. Colonization factor antigen II receptor activity of mannose-resistant hemagglutination-positive erythrocytes is unstable when whole blood is stored at 4 degrees C. Also, screening of donors is best performed with enterotoxigenic E. coli possessing colonization factor antigen II composed of the coli surface antigen 1 (CS1) plus CS3, since these consistently produce stronger hemagglutination reactions than strains with colonization factor antigen II composed of either CS2 plus CS3 or CS3 only. PMID- 3053771 TI - Characterization of Candida isolates from pediatric burn patients. AB - To provide more detailed information about Candida epidemiology and pathogenesis in pediatric burn patients, Candida isolates from 113 patients collected over 3 years were identified at the species level and the serotypes and biotypes of the C. albicans isolates were determined. A total of 85% of the patients were colonized or infected by C. albicans, 18% by C. tropicalis, and 11% by C. parapsilosis. Although colonization or infection often was found at multiple sites and times, 87% of the patients were colonized or infected by only one Candida species or strain; the other 13% showed multiple colonizations or infections, some of which occurred simultaneously at the same site. C. albicans biotyping determined the tolerance of the isolates to pH (pH 1.4) and salt; flucytosine, borate, and safranine resistance; and ability to produce proteinase and assimilate urea, sorbose, and citrate; results are expressed as three-digit numbers. For isolates from three different anatomical sites, the distribution of the nine biotype characteristics was similar in all cases but one. Significantly more fecal than wound or throat isolates were resistant to safranine. Sixty-four different serotype-biotype combinations were found in the 96 patients with C. albicans infections or colonizations. Twenty-nine percent of all C. albicans isolates had the partial biotype -57, while 20 of the 96 patients had specifically serotype B, biotype 557 colonizations or infections. Eleven patients had the B557 infection when admitted; nine patients acquired the yeast in-house. Thirty percent of the C. albicans isolated from 23 adult patients at a nearby hospital also showed the -57 biotype pattern, suggesting that C. albicans isolates expressing this biotype are either extremely prevalent in nature or are more virulent than other C. albicans isolates. PMID- 3053772 TI - Enantioselective measurement of fungal D-arabinitol in the sera of normal adults and patients with candidiasis. AB - A new method was used to measure D-arabinitol enantioselectively in the sera of 27 healthy adults and four patients with candidiasis. Arabinitol was measured by gas chromatography in serum that was treated with and without the Klebsiella pneumoniae enzyme D-arabinitol dehydrogenase, lactic dehydrogenase, NAD, and sodium pyruvate. Since enzyme treatment removed 98% of 0 to 20 micrograms of D arabinitol per ml and none of 0 to 20 micrograms of L-arabinitol per ml from spiked sera, D-arabinitol could be determined from the difference in the treated and untreated samples. The concentrations of D- and L-arabinitol in serum from normal subjects were 0.22 +/- 0.052 and 0.16 +/- 0.055 micrograms/ml, respectively, and their D-arabinitol/creatinine and L-arabinitol/creatinine ratios were 0.024 +/- 0.0089 and 0.017 +/- 0.0053 (all means +/- standard deviations). The infected patients all had markedly elevated serum D-arabinitol levels, but their L-arabinitol levels were either normal or proportionately much lower. The excess arabinitol in the sera of individuals with candidiasis is D arabinitol, and use of enantioselective analytical methods should result in improved ability to diagnose and estimate the severity of candidiasis. PMID- 3053773 TI - Legionella pneumophila serogroup Lansing 3 isolated from a patient with fatal pneumonia, and descriptions of L. pneumophila subsp. pneumophila subsp. nov., L. pneumophila subsp. fraseri subsp. nov., and L. pneumophila subsp. pascullei subsp. nov. AB - Previous DNA relatedness and enzyme electrophoretic mobility studies indicated heterogeneity among strains of Legionella pneumophila serogroups 1, 4, 5, and Lansing 3 (a new, as yet unnumbered serogroup). In this study 60 L. pneumophila strains were studied by DNA hybridization (hydroxyapatite method) to assess their genomic relatedness. These strains were also studied biochemically and serologically to determine whether they formed one or more phenotypic groups. DNA relatedness studies identified three groups. DNA group 1 contained the type strain Philadelphia 1 and strains from serogroups 1 through 14 of L. pneumophila. The average relatedness of DNA group 1 strains was 88% at 60 degrees C with 1.1% divergence in related sequences and 85% at 75 degrees C. DNA group 2 contained strain Los Angeles 1, the reference strain of serogroup 4, and strains of serogroups 1, 4, 5, and Lansing 3, an unnumbered serogroup. Average relatedness of DNA group 2 strains was 84% at 60 degrees C with 0.7% divergence and 87% at 75 degrees C. Reciprocal relatedness of DNA groups 1 and 2 was approximately 67% at 60 degrees C with 6.0% divergence and 48% at 75 degrees C. DNA group 3 strains were in serogroup 5. They were 98% related at 60 degrees C with 0.5% divergence and 97% related at 75 degrees C. Reciprocal relatedness of DNA group 3 and DNA group 1 was approximately 74% at 60 degrees C with 5.3% divergence and 43% at 75 degrees C, and reciprocal relatedness of DNA groups 3 and 2 was 66% at 60 degrees C with 5.7% divergence and 55% at 75 degrees C. The DNA groups could not be separated biochemically or serologically or by cell wall fatty acid and isoprenoid quinone composition. Three subspecies of L. pneumophila are proposed to accommodate the three DNA groups: L. pneumophila subsp. pneumophila subsp. nov. for DNA group 1, L. pneumophila subsp. fraseri subsp. nov. for DNA group 2, and pneumophila subsp. pascullei subsp. nov. for DNA group 3. PMID- 3053777 TI - Enterococcus hirae implicated as a cause of diarrhea in suckling rats. AB - A Lancefield group D enteric streptococcus was isolated from diarrheic suckling rats that had been inoculated orally with stool from a diarrheic human. After oral administration of the organism to other suckling rats, diarrhea was reproduced, and the enteric streptococcus was reisolated. The brush border of small intestinal villi in affected animals was coated with numerous adherent gram positive cocci. The organism was identified as Enterococcus hirae by a battery of biochemical tests. These and previous studies indicate that certain enterococci should be considered as etiologic agents of diarrheal disease in neonatal animals. PMID- 3053774 TI - DNA hybridization for assessment of response of Plasmodium falciparum to chloroquine therapy. AB - We studied the accuracy of PFR1-AP, a synthetic DNA hybridization probe conjugated to alkaline phosphatase, in monitoring Plasmodium falciparum parasitemia during in vivo drug susceptibility surveys. Duplicate blood samples were collected from six children enrolled in a 14-day in vivo chloroquine study in Rwanda. Results obtained by microscopic examination of Giemsa-stained thick blood smears and by DNA hybridization were compared. Both techniques successfully monitored an infection with chloroquine-susceptible parasites and infections exhibiting various levels of resistance to treatment. For each patient, temporal evolution of the microscopic parasite counts and the DNA hybridization signals were closely parallel, although a wide range of rapidly changing levels of parasitemia occurred through the course of the study. This suggests that DNA hybridization assay using PFR1-AP detects P. falciparum parasites sensitively and specifically and is a valuable tool for drug resistance surveys. PMID- 3053776 TI - Restriction endonuclease analysis of Staphylococcus epidermidis DNA may be a useful epidemiological marker. AB - We compared the epidemiological markers of 13 Staphylococcus epidermidis strains isolated from an adult inpatient during a febrile episode and 23 S. epidermidis strains isolated during a presumptive outbreak of nosocomial infection in a neonatal ward. The total DNA restriction endonuclease analysis (REA) was processed along with the following conventional markers: biotyping, serotyping, phage typing, antibiotic susceptibility profiles, and plasmid profiles. The REA method was reproducible, giving stable results both in vitro and in vivo. For the hospitalized adult patient, the conventional markers of the 13 strains were concordant and the restriction profiles were identical. Five restriction groups were demonstrated during the course of the outbreak. Within two of the groups, the identities of all of the markers were used to verify whether all of the isolates belonged to the same cell clone. In a third group, combined analysis of the conventional markers and REA had to be used to demonstrate isolate similarity. On the other hand, in another group, none of the markers were similar; interpretation was not easy. An epidemiological study of S. epidermidis infections in hospitals must take into account all of the epidemiological markers: biotypes, serotypes, phage types, antibiograms, plasmid profiles, and REA. PMID- 3053775 TI - Humoral immune response in human tuberculosis: immunoglobulins G, A, and M directed against the purified P32 protein antigen of Mycobacterium bovis bacillus Calmette-Guerin. AB - The P32 protein antigen of Mycobacterium bovis BCG, identified as antigen 85A in the BCG reference system, was used to investigate the humoral immune response in human tuberculosis (TB). Immunoglobulin G (IgG), IgA, and IgM directed against P32 were measured by an enzyme-linked immunosorbent assay. Mean IgG and IgA antibody levels differed significantly (P less than 0.001) between active-TB patients (50 untreated and 52 treated) and healthy control subjects (111 unvaccinated tuberculin negative, 38 unvaccinated tuberculin positive, and 72 recently BCG vaccinated). Mean IgG antibody levels, but not mean IgA antibody levels, were higher (P less than 0.05) in patients with positive microscopic examination for acid-fast bacilli than in patients with negative microscopic examination. A positive relation was found between mean levels and the extent of disease. There was no difference in mean IgM antibody levels between patients and controls. By setting the upper normal limit at the 95th percentile of the 221 healthy subjects, the sensitivities were 46% in untreated and 63% in treated patients for IgG and 30 and 50%, respectively, for IgA. Of the untreated patients, 56% were positive for either IgG or IgA antibodies. Among the untreated patients with negative direct smear, 35% were positive for IgG and 24% were positive for IgA. When both immunoglobulin classes were combined, the serological test was positive in 47% of those patients. Neither naturally acquired tuberculin hypersensitivity nor BCG vaccination affected positivity frequencies in healthy subjects. Only active TB seemed to induce significant anti-P32 antibody levels and to be associated with positivity. A serological test with P32 as the antigen might therefore be helpful for the rapid diagnosis of TB. PMID- 3053779 TI - Identification of Actinomyces (Corynebacterium) pyogenes with the API 20 Strep system. AB - A total of 62 strains of Actinomyces pyogenes (previously Corynebacterium pyogenes) were examined by the API 20 Strep system (API System, La Balme Les Grottes, Montalieu-Vercieu, France). The system was shown to be reliable and rapid when the tests were compared with standard identification methods. No confusion occurred with streptococcal profiles in the current API 20 Strep data base. PMID- 3053778 TI - Detection of Vibrio cholerae with monoclonal antibodies specific for serovar O1 lipopolysaccharide. AB - Six hybridoma cell lines, each of which produced a monoclonal antibody (MAb) against Vibrio cholerae O1 lipopolysaccharide (LPS), were established. Each MAb was active serologically by both enzyme-linked immunosorbent assay (ELISA) and the slide agglutination test. In the ELISA, each MAb was tested against 7 O1 and 9 non-O1 LPS preparations. Three MAbs reacted with both Inaba and Ogawa serovars (A antigen), two MAbs reacted with the Ogawa serovars only (B antigen), and one MAb reacted with the Inaba serovars only (C antigen). Each MAb was also tested in the ELISA against whole-cell preparations of 37 O1 and 52 non-O1 V. cholerae serovars, 20 heterologous Vibrio species, and 37 heterologous bacterial species. The MAbs reacted with V. cholerae O1 cells only, except for one anti-A antigen MAb which reacted weakly with five V. cholerae non-O1 serovars and Serratia marcescens. Each anti-A antigen MAb was labeled with fluorescein isothiocyanate (FITC) and tested by direct immunofluorescence against selected O1 and non-O1 serovars. Each MAb-FITC conjugate, when tested alone, exhibited O1-specific fluorescence; however, mixtures of the MAb-FITC dramatically enhanced fluorescence intensity on O1 cells. This finding was also visualized by immunoelectron microscopy on both thin-sectioned and negatively stained O1 cells by using an anti-mouse immunoglobulin-colloidal gold conjugate. These results suggest that the A antigen can be described by more than one epitope and that a superior serotyping reagent can be prepared from a defined mixture of MAbs. PMID- 3053780 TI - Detection of bacteria in the presence of antibiotics by using specific monoclonal antibodies to neutralize the antibiotics. AB - Inactivation of penicillin and gentamicin in cultures was achieved by using monoclonal antibodies against these antibiotics. A viridans group streptococcus (penicillin MIC, less than or equal to 0.06 micrograms/ml) and Escherichia coli ATCC 35218 (gentamicin MIC, less than or equal to 1 microgram/ml) were able to grow in broth containing 0.25 micrograms of penicillin per ml and 4 micrograms of gentamicin per ml, respectively, when the specific antibodies were added. This procedure may be useful to increase the yield of bacteria from body fluid specimens that contain antibiotics. PMID- 3053782 TI - An immunoelectron microscopical study of the expression of class II MHC and a T lymphocyte surface marker during chronic relapsing experimental allergic encephalomyelitis. AB - Immunoelectron microscopy using antibodies recognising Class II MHC antigens and a pan T cell marker was employed to study sections of spinal cord from guinea pigs with chronic relapsing experimental allergic encephalomyelitis (CREAE). It was found that endothelial cells expressed Class II antigens on their luminal surface throughout the course of the disease and that lymphocytes were adherent to these surfaces. In the parenchyma lymphocytes, macrophages and possibly microglia expressed Class II antigens suggesting that they might also be involved in antigen presentation. The different distribution of T cells seen in the individual lesions during the relapse phase may correlate with their respective natural histories. PMID- 3053783 TI - Contribution of electrophysiological studies to cerebellar physiology. AB - Electrical stimulation and the recording of electrical potentials have made important contributions to the classic formulations of cerebellar function. These electrical methods also have contributed to a re-appraisal, now underway, of the cerebellar role in the human brain. Beyond its accepted role in motor function, the cerebellum seems to contribute to some nonmotor functions. It is now known to communicate with the prefrontal cortex as well as with the motor cortex of the frontal lobe, and it seems to be involved in some prefrontal cognitive and language functions as well as in motor function. Investigations of cerebellar involvement in such functions are now being carried out, with encouraging results. If confirmed by further clinical evidence, this broader concept of cerebellar function would explain the mystery of why the most lateral parts of the cerebellum enlarged dramatically in the human brain, concomitantly with the enlargement of the cerebral association areas. PMID- 3053781 TI - Solid-phase immunosorbent technique for rapid detection of Rift Valley fever virus immunoglobulin M by hemagglutination inhibition. AB - A solid-phase immunosorbent technique (SPIT) was adapted to detect Rift Valley fever (RVF) virus-specific immunoglobulin M (IgM) in serum samples from humans vaccinated with Formalin-inactivated RVF vaccine. Microdilution plates coated with goat anti-human IgM were successively incubated with serum samples from human vaccinees, RVF virus hemagglutinating antigen, and goose erythrocytes. The RVF virus-specific IgM in the serum samples from vaccinees bound to the RVF virus antigen and inhibited hemagglutination of goose erythrocytes. SPIT was compared to the IgM capture enzyme linked immunosorbent assay (ELISA) and the indirect immunofluorescent-antibody (IFA) assay and was found to be sensitive in detecting RVF virus-specific IgM antibody, with high correlations between SPIT and the other two tests (Pearson's correlation coefficient [r] = 0.9 and 0.6, respectively). Results of SPIT were obtained within 5 h, offering speed over ELISA (8 h). In addition, SPIT does not require sophisticated equipment or expensive reagents. Serum rheumatoid factor did not produce false-positive reactions in SPIT as in the indirect immunofluorescent-antibody assay and IgM capture ELISA. PMID- 3053787 TI - Clinical and microbiological effects of root debridement in periodontal furcation pockets. AB - The aim of the present study was to investigate longitudinally over 52 weeks the clinical and microbiological effects of plaque control and root debridement at molar furcation sites. The results were compared with changes at non-molar sites. 24 non-molar sites and 31 grade II molar furcation sites with probing depth greater than or equal to 5.0 mm were monitored in 11 patients. Clinical measurements consisted of plaque scores, probing depths, and changes in probing attachment level. Microbiological monitoring was carried out with phase-contrast microscopy and anaerobic culturing. The debridement resulted in improvement in probing measurements and microbiological counts for both groups of sites. A slightly less favorable clinical response was noted for molar furcation sites. Higher post-operative microbiological counts were found throughout the 52-week observation period for molar furcation sites. Sites with probing attachment loss showed higher microbial counts and higher proportions of spirochetes, black pigmented colony forming units (CFU), and Bacteroides gingivalis CFU than sites with probing attachment gain. Individual site analysis, however, demonstrated marked variations of the microbiological counts at the different postoperative time points. In the few available sites undergoing probing attachment loss, no apparent association between target micro-organisms and periodontal deterioration was observed. PMID- 3053786 TI - Detection of high-risk groups and individuals for periodontal diseases. Clinical assessment of the periodontium. AB - The fundamental concepts of measuring periodontal diseases and the interpretation of such information as an historical record of disease, rather than disease activity, emphasises the need for improved diagnostic and prognostic tests. Criteria for an indicator of disease activity were suggested and an index fulfilling these should allow sites to be categorised as "active", quiescent or healing, and enable one to predict the risk of future disease activity. The ability of current measurements and indices, routinely used during clinical assessments of periodontal diseases, to fulfill the suggested criteria was considered and rejected in all cases. It is concluded that clinical parameters are only capable of identifying disease retrospectively, indicating the need for longitudinal, rather than cross-sectional studies in the search for clinical and laboratory markers of disease activity and susceptibility. PMID- 3053784 TI - A malarial cysteine proteinase is necessary for hemoglobin degradation by Plasmodium falciparum. AB - To obtain free amino acids for protein synthesis, trophozoite stage malaria parasites feed on the cytoplasm of host erythrocytes and degrade hemoglobin within an acid food vacuole. The food vacuole appears to be analogous to the secondary lysosomes of mammalian cells. To determine the enzymatic mechanism of hemoglobin degradation, we incubated trophozoite-infected erythrocytes with peptide inhibitors of different classes of proteinases. Leupeptin and L transepoxy-succinyl-leucyl-amido-(4-guanidino)-butane (E-64), two peptide inhibitors of cysteine proteinases, inhibited the proteolysis of globin and caused the accumulation of undegraded erythrocyte cytoplasm in parasite food vacuoles, suggesting that a food vacuole cysteine proteinase is necessary for hemoglobin degradation. Proteinase assays of trophozoites demonstrated cysteine proteinase activity with a pH optimum similar to that of the food vacuole and the substrate specificity of lysosomal cathepsin L. We also identified an Mr 28,000 proteinase that was trophozoite stage-specific and was inhibited by leupeptin and E-64. We conclude that the Mr 28,000 cysteine proteinase has a critical, perhaps rate-limiting, role in hemoglobin degradation within the food vacuole of Plasmodium falciparum. Specific inhibitors of this enzyme might provide new means of antimalarial chemotherapy. PMID- 3053788 TI - Role of "diseased" root cementum in healing following treatment of periodontal disease. A clinical study. AB - This clinical trial was undertaken to examine whether root debridement in the treatment of periodontal disease must include the removal of the exposed cementum in order to achieve periodontal health. The study included 11 adult patients with moderate to advanced periodontal disease. In a split-mouth design, the dentition of each patient was by random selection divided into test- and control quadrants comprising the incisors, canines and premolars. Following a baseline examination, all patients were given a case presentation and a detailed instruction in self performed oral hygiene measures. The patients were then subjected to periodontal surgery. Following reverse bevel incisions, buccal and lingual mucoperiosteal flaps were elevated and all granulation tissue was removed. In 2 jaw quadrants (control quadrants) in each patient, the denuded root surfaces were carefully scaled and planed in order to remove soft and hard deposits as well as all cementum, using hand instruments and flame-formed diamond stones. In the contralateral quadrants (test quadrants) the roots were not scaled and planed but soft microbial deposits were removed by polishing the root surfaces with the but soft microbial deposits were removed by polishing the root surfaces with the use of rubber cups, interdental rubber tips and a polishing paste. Calculus in the test quadrants was removed by the use of a curette, but precaution was taken to avoid the removal of cementum. The flaps were repositioned to their original level and sutured. The patients were following active treatment enrolled in a supervised maintenance care program including "professional tooth cleaning" once every 2 weeks for a 3-month period.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3053785 TI - Synergistic regulation of human megakaryocyte development. AB - Little information exists concerning differing levels of regulation occurring during human megakaryocyte development. We hypothesize that megakaryocytic proliferation and maturation is controlled by two, synergistic regulatory factors. One, megakaryocyte colony-stimulating activity, is an obligate requirement for colony formation and drives the development of relatively immature cells. Megakaryocyte colony-stimulating activity is a functional component of the human recombinant proteins, interleukin 3 or GM-CSF. Human recombinant growth factors, interleukin 1, interleukin 6, or crythropoietin, do not effect megakaryocyte development either alone or in combination with interleukin 3. Full maturation requires a second synergistic activity which increases megakaryocyte number, size, and cytoplasmic and antigenic content. In culture, this synergistic regulator augments maturation by increasing the number of colonies, colony cellularity, and size. In suspension cultures, this cofactor increases megakaryocyte cytoplasmic and antigenic content, and shifts the morphological distribution from immature to mature megakaryocytes. Finally, this activity also increases the number of antigen positive megakaryocytes, either by stimulating proliferation or conversion of antigen-negative to antigen-positive cells. Comparative studies of megakaryocytic regulation suggests that this in vitro regulator mimicks some of the known effects of thrombopoietin in vivo. PMID- 3053789 TI - The role of dental plaque in gingivitis and periodontitis. AB - A dynamic equilibrium between the periodontal microbiota and the host generally results in a clinical state of periodontal health, characterized by minimal inflammatory changes in the marginal gingival tissues. Maintenance of health is most easily achieved by controlling the resident mass of bacteria. In rare instances, control of specific microorganisms may be indicated. Lack of microbial control may lead to an imbalance between the microbiota and the host due to a markedly increased microbial mass and/or increased virulence of the micro organisms present. Such alterations in the host-parasite equilibrium may result in transient episodes of tissue destruction and, in the long term, to cumulative damage to the periodontal tissues. PMID- 3053790 TI - Chemotherapeutic agents for controlling plaque and gingivitis. AB - There has been a vigorous search for many years for chemical agents that could supplement or even supplant patient-dependent mechanical plaque control and thus reduce or prevent oral disease. 5 categories of agents or approaches have been considered: (1) broad spectrum antiseptics, (2) antibiotics aimed at specific bacteria, (3) single or combinations of enzymes that could modify plaque structure or activity, (4) non-enzymatic dispersing or modifying agents and (5) agents that could affect bacterial attachment. The success of these approaches can be evaluated clinically by the use of standard scoring methods for measuring plaque and gingivitis and their safety established by soft tissue and microbiologic examination. Antiseptic agents have received the bulk of the attention over the years. At present, only 2 antiseptics, the bis-biguanide, chlorhexidine gluconate (Peridex) and a combination of phenol related essential oils (Listerine), have developed sufficient supporting data in 6-month (or longer) studies to gain the approval of the Council On Dental Therapeutics of the American Dental Association. On the basis of short-term studies, cetylpyridinium chloride, zinc and copper salts, sanguinarine and octenidine warrant continued study as does stannous fluoride at an appropriate concentration. On the basis of current research, a new generation of more specific antibacterial agents that interfere with attachment to pellicle can be developed. It is hard to predict, however, that they will affect gingivitis, at least until there is more information on what specific organisms should be targeted. PMID- 3053791 TI - Microbiological effects of mouthrinses containing antimicrobials. AB - A number of mouthrinse formulations containing antimicrobials have been evaluated to determine their effectiveness as antiplaque and/or antigingivitis agents. These have included the bis-biguanides, phenols, quaternary ammonium compounds, oxygenating compounds, plant extracts, fluorides, antibiotics and antimicrobial combinations. These mouthrinses have often been tested as adjuncts to normal oral hygiene procedures as well as in the experimental gingivitis model. 2 agents in particularly, chlorhexidine gluconate and listerine, have been shown to both inhibit or reduce plaque accumulation and the severity of gingivitis. Chlorhexidine has been reported to reduce the accumulation of plaque by approximately 60% and the severity of gingivitis by 50-80% as determined by improvements in clinical indices. A 0.12% chlorhexidine gluconate rinse resulted in significant reductions after both 3 and 6 months use in the numbers of total anaerobes, total aerobes, streptococci, and actinomyces recovered from supragingival plaque. Listerine has been reported to retard the development of plaque by 45 to 56% and to reduce existing plaque by 39 to 48%. Gingivitis scores were reduced as much as 59%. Microbial studies have shown that the effect of listerine is exerted against the total microbial mass and results in an overall decrease in both the biomass and the activity. Long-term use of neither mouthrinse, chlorhexidine or listerine, resulted in the emergence of opportunistic or oral pathogens. Preliminary data obtained following the use of a novel mouthrinse consisting of a combination of povidone-iodine and hydrogen peroxide appears promising. This combination was more effective than was more effective than either single component alone in reducing gingivitis scores.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3053792 TI - Clinical index systems used to assess the efficacy of mouthrinses on plaque and gingivitis. AB - The American Dental Association's Council on Dental Therapeutics has adopted Guidelines for acceptance of chemotherapeutic products for the control of supragingival dental plaque and gingivitis. The most widely used plaque indices are the plaque index (PI) and the Turesky index. Gingivitis has usually been assessed by the Loe and Silness method, although the modified gingival index of Lobene and a bleeding index reported by Caton and Polson have also been used. To date, 2 products have been accepted by the Council as being effective in helping control supragingival plaque and gingivitis. These products were evaluated using clinical indices described in this review. The indices were selected from the many in the literature as being valid, reliable, and easily learned. Suggestions are made for criteria to be used in comparability studies. PMID- 3053795 TI - Use of mouthrinses for professional indications. AB - In view of the relationship between supragingival and subgingival plaque, chemical agents which alter supragingival plaque may also alter subgingival plaque. As a result, a beneficial effect on gingival health may be anticipated. This article reviews the use of chemotherapeutic agents which reduce plaque and gingivitis. Indications for the use of these agents include patients with problems with mechanical plaque control, extensive splinting or fixed prostheses, intraoral fixation, orthodontic appliances, overdenture abutments and implants, and for patients in the immediate post-surgical period. Improvement of wound healing following periodontal surgery may represent one of the most valuable aspects of use of these agents. As mouthrinses, these agents cannot be expected to significantly alter subgingival plaque. However, for this purpose, their use in irrigation devices deserves further study. Since recent reports have suggested benefits of various mouthrinses when used as irrigants as compared to rinses may also reduce the bacterial back-spray associated with ultrasonic devices and high speed handpieces, and thus provide a protective effect for dental personnel. The practitioner in the United States is encouraged to recommend products accepted by the American Dental Association's Council on Dental Therapeutics in order to insure both safety and efficacy. PMID- 3053793 TI - Mouthrinses in "experimental gingivitis" studies. AB - The experimental gingivitis study design is a frequently used clinical model for the evaluation of the effects of antimicrobial agents on developing plaque and gingivitis. In individuals who at the start of the experiment have clean teeth and healthy gingivae and who use a mouthrinse containing the test agent as the only tooth cleaning measure during a period of 14-21 days, the anti-plaque and anti-gingivitis potential of the agent can be properly evaluated. The present paper describes different designs of experimental gingivitis studies for the evaluation of chemotherapeutic agents used to control supragingival plaque formation. Findings reported in the literature with respect to the effects obtained by various antimicrobial compounds are also reported. PMID- 3053794 TI - Longitudinal clinical studies with antimicrobial mouthrinses. AB - This chapter examines the use of long-term clinical studies to evaluate the effectiveness of antimicrobial mouthrinses in the prevention and/or treatment of plaque and gingivitis. Such studies should be double-blind, compare the active agent to a control, and last a minimum of 6 months. Such a time interval simulates a common recall interval, enables one to utilize a thorough dental prophylaxis at the initiation of the study, and is sufficient time to observe the development of toxic and other side-effects of therapy. The microbiological component should estimate plaque quantitatively and demonstrate that no pathogenic, opportunistic or resistant micro organisms develop. The indices and microbiology should be performed at baseline, and intermediate period and at 6 months. Thus far, published studies meeting these criteria support the efficacy of listerine and chlorhexidine as agents useful in reducing plaque and gingivitis. Preliminary evidence indicates that sanguinarine may also be effective. The most common side effect of listerine was the complaint of poor taste. Increased stain and calculus formation were cited as the most significant side-effects of chlorhexidine. Such side-effects must be taken into consideration when recommending effective antimicrobial mouthrinses for patient use. PMID- 3053797 TI - Weight gain with antidepressants and lithium. AB - Undesired weight gain is a common complaint of patients receiving pharmacological treatment for major affective disorders. It has been found to jeopardize patient compliance and may pose additional health hazards. A review of the literature on weight gain associated with tricyclic antidepressants, monoamine oxidase inhibitors, and lithium was carried out with the aim of deriving practical management strategies. Tricyclic antidepressants were found to stimulate appetite, carbohydrate craving, and a dose-dependent continuous weight gain of 0.57 to 1.37 kg per month of treatment. Proposed mechanisms include noradrenergic or antihistaminic inhibition of satiety and decreased metabolic rate. Novel serotonergic and dopaminergic antidepressants were found to be anorectic. Monoamine oxidase inhibitors may stimulate appetite and potentiate insulin induced hypoglycemia. Lithium maintenance therapy stimulates weight gains of over 10 kg in 20% of patients. Documented mechanisms include insulin-like actions on carbohydrate and fat metabolism, polydipsia, and sodium retention. Recommendations regarding choice of antidepressant drug as well as dietary and behavioral strategies to prevent excessive weight gain are presented. Potential adjunctive drug approaches to severe weight gain are reviewed. PMID- 3053796 TI - Antidepressant drug studies, 1964 to 1986: empirical evidence for aging patients. AB - This review is based on 25 double-blind anti-depressant drug studies reported between 1964 and 1986 that focused on patients over 55 years of age. The number of studies located in the literature is appallingly small, particularly when we consider that we included experimental drugs as well as drugs not recommended for use with the elderly. In general, the results of our survey support clinical experience: the drugs are clearly superior to placebo; they show comparable therapeutic efficacy--about 50% improvement in Hamilton Psychiatric Rating Scale for Depression scores versus 20% to 25% on placebo; and all of them have undesirable side effects. Thus, the choice of drug is based on side effects profiles and potential drug-drug interactions rather than on degree of therapeutic efficacy. The review makes apparent the need for more substantial data on treatment outcome in patients over the age of 60 years. Beyond that, additional information is unlikely to change the 50% response rate or the potential for serious side effects with most of the currently available drugs. We clearly need better drugs. PMID- 3053798 TI - Management strategies for developmental apraxia of speech: a review of literature. AB - In recent years, interest in and controversy about developmental apraxia of speech has been abundant. This paper is designed to provide a series of management strategies for developmental apraxia of speech. There are numerous unanswered questions about the effects of therapy; pertinent research issues are identified. PMID- 3053799 TI - Prevention of communication problems associated with cleft palate. AB - The purpose of this paper is to review principles of preventative intervention and their application to communication problems associated with cleft palate. Ten specific suggestions and activities are described. PMID- 3053800 TI - Local anesthesia in dermatologic surgery. AB - With the growth of dermatologic surgery the appropriate use of local anesthesia is becoming an important issue. Thoughtful use of local anesthesia can improve the patient's experience and facilitate the surgical procedure. In this review we discuss historical and pharmacologic aspects of local anesthetic agents. Emphasis is placed on clinical considerations, including contraindications, toxic reactions, and detailed descriptions of anesthetic use. PMID- 3053801 TI - Classification of peristomal skin changes in patients with urostomy. AB - Peristomal skin lesions in patients with ileal conduit urinary diversion have been reported in frequencies ranging from none to occurrence in 100% of patients. In previous studies skin lesions often are referred to in unspecific terms, which has made it impossible to compare results with those of other studies. Thus a classification of peristomal skin (CPS) has been developed on the basis of macroscopic peristomal findings. It defines what can be accepted as ordinary findings, as well as two different types of skin lesions, erythematous-erosive and pseudoverrucous. Each of these is divided into two subgroups. Such a classification is a prerequisite for a meaningful comparison of the type, incidence, and severity of skin lesions in different groups; it also may allow a more reliable clinical evaluation of new ostomy appliances and skin care products. In addition, CPS may also facilitate communication among professionals who are responsible for the care of ostomy patients. PMID- 3053802 TI - Contact immunotherapy of resistant warts. AB - Contact immunotherapy has been proved effective in the treatment of resistant warts. This report chronicles our experience with a new contact immunotherapy agent, diphenylcyclopropenone. We have achieved a cure rate of 62% in 45 patients with resistant warts of all types who came to our general dermatology clinic. Cure rates may be lower in patients who have experienced multiple treatment failures. The majority of cures were obtained within 3 to 4 months. Although it appears somewhat less effective than published reports of dinitrochlorobenzene contact immunotherapy, diphenylcyclopropenone contact immunotherapy is an effective treatment for resistant warts and avoids any potential problems from mutagenicity. PMID- 3053804 TI - Malignant blue nevus. Case report and literature review. AB - A well-documented case of malignant blue nevus is presented, along with an in depth review of the literature. Malignant blue nevus is a rare form of malignant melanoma. A cellular blue nevus is the precursor lesion. The scalp is the most common site. The tumor often presents clinically as a progressively enlarging or multinodular blue-black lesion. The histologic pattern is fascicular dense collections of pigmented, pleomorphic spindle cells. Because of marked regional histologic variation within a malignant blue nevus, however, sampling error can cause delay in recognition of malignancy. A high clinical index of suspicion and appropriate biopsy technique are necessary to reach an early diagnosis. The most common site of metastasis of a malignant blue nevus is the lymph node. The phenomenon of benign lymph node nevus cell metastasis, which may occur with benign blue nevi, must be differentiated from a true malignant metastasis of a malignant blue nevus. PMID- 3053803 TI - Pachyonychia congenita. AB - Pachyonychia congenita is a rare hereditary disorder characterized mainly by nail hypertrophy and dyskeratoses of skin and mucous membranes. A thorough literature survey since its first description in 1904 up to 1985 revealed 168 cases of pachyonychia congenita. There were no indications of any sex or ethnic group predilection. Based on this survey the following classification is suggested: type I (56.2% of cases), hyperkeratosis of nails, palmoplantar keratosis, follicular keratosis, and oral leukokeratosis; type II (24.9% of cases), clinical findings of type I plus bullae of palms and soles, palmar and plantar hyperhidrosis, natal or neonatal teeth, and steatocystoma multiplex; type III (11.7% of cases), clinical findings of types I and II plus angular cheilosis, corneal dyskeratosis, and cataracts; and type IV (7.2% of cases), clinical findings of types I, II, and III plus laryngeal lesions, hoarseness, mental retardation, hair anomalies, and alopecia. PMID- 3053806 TI - Computed tomography of fatal cerebral air embolism following percutaneous aspiration biopsy of the lung. AB - Air embolism complicated a thin needle aspiration performed on a patient with adult respiratory distress syndrome and on positive-pressure ventilation. Computed tomography obtained 30 h following the event demonstrated a considerable quantity of intravascular air within the cranium. Positive-pressure ventilation should be considered a relative contraindication for thin needle lung aspiration. PMID- 3053805 TI - Micrographic surgery for the microscopically controlled excision of carcinoma of the external ear. AB - Micrographic surgery is particularly valuable for treating carcinomas of the external ear because the total microscopic control of excision afforded by this method permits selective removal of the clinically unpredictable cancerous outgrowths. The maximum conservation of normal tissues, including the cartilaginous framework that gives shape to the ear, is a benefit next in importance to the unusually high 5-year cure rates (97.1% in a series of 1213 cases of basal cell carcinoma and 92.3% in a series of 871 cases of squamous cell carcinoma). The fresh-tissue technique for removing most auricular carcinomas usually permits the excision of the successive layers in 1 day, and repair, if needed, can be done immediately. The fixed-tissue technique is still used by the authors for some extensive, bone-invading carcinomas. PMID- 3053810 TI - Computed tomography of urachal carcinoma. AB - Urachal carcinoma arises in the clinically silent extraperitoneal space between the bladder apex and the umbilicus. Computed tomography was used to correctly diagnose and stage urachal cancer in two patients. In one case a mass was localized to the bladder wall and immediate juxtavesical region; in the other case an advanced locally invasive lesion was seen to engulf and fisulize loops of small bowel and extend through the umbilicus. Urachal carcinoma has a highly characteristic appearance and location on CT images. PMID- 3053807 TI - MR body stereotaxis: an aid for MR-guided biopsies. AB - The use of a new body stereotactic system to facilitate magnetic resonance (MR) guided biopsies is described. A skin entry point is first found using a localizer MR scan. The articulating arm of the stereotactic unit is then used to aim the MR needle at the entry point and accurately maintain the needle at the correct angle to intersect the target area. Complex angles may be used to allow needle passes outside the scan plane. This is accomplished by angling the arm out of the plane of the section. PMID- 3053809 TI - CT demonstration of retrohepatic gallbladder in severe cirrhosis. AB - In two patients with hepatic cirrhosis and marked morphological distortion of the liver, the gallbladder was identified in a retrohepatic position on ultrasound and CT. The etiology of this anomalous position and the importance of the preoperative diagnosis of both congenital and acquired anomalous position of the gallbladder in patients with acute cholecystitis is discussed. PMID- 3053811 TI - Recognition of malignant melanoma by monoclonal antibody HMB-45. An immunohistochemical study of 200 paraffin-embedded cutaneous tumors. AB - Antibodies to S-100 protein have been used widely as markers of malignant melanoma, despite abundant evidence that they are non-specific for this neoplasm. Hence, alternatives to these reagents are desirable in diagnostic dermatopathology. We evaluated the characteristics of a new monoclonal antibody (HMB-45) which does have putative specificity for melanoma, and compared it with a polyclonal anti-S-100 reagent in immunohistochemical staining of 67 melanomas of the skin and 133 non-melanomatous cutaneous neoplasms. All specimens were formalin-fixed and paraffin-embedded, and were studied with the avidin-biotin peroxidase complex technique. HMB-45 labelled 62 of 67 melanomas, while anti-S 100 recognized all tumors of this type. On the other hand, S-100 also was expressed by 15 of the non-melanocytic neoplasms, all 133 of which were HMB-45 negative. The only cases of melanoma that were missed by the latter reagent were of the spindle-cell type. Hence, HMB-45 was 100% specific and 93% sensitive, relative to a diagnosis of malignant melanoma in paraffin sections. Epithelioid and small-cell neoplasms are reliably recognized by this antibody, but it would appear that spindle-cell melanomas must be detected by other immunohistochemical means. PMID- 3053808 TI - Cerebral venous angiomas: MR findings. AB - Seven patients with suspected cerebral venous angioma studied by either CT or angiography were imaged with magnetic resonance. Six of seven cases demonstrated a stellate appearance on contrast enhanced CT. In two patients this finding was verified by angiography. Flow void was identified on both T1- and T2-weighted pulse sequences. In one patient a field echo sequence demonstrated high intensity signal within the venous angioma. Magnetic resonance proved superior to CT in the identification of these lesions. A stellate configuration with an emanating transcortical vein and centrifugal drainage (transcortical venous flow) from the angioma into a sinus was present in all cases. Centripetal drainage via thalamostriate and internal cerebral veins was not seen. There was no evidence of mass effect, scar, or hemorrhage. Four of the angiomas were located in a frontal lobe and three in a cerebellar hemisphere. This distribution of the lesions is similar to that reported in the literature in which the frontal lobe is the most common location followed by the cerebellar hemisphere. An embryological explanation is cited and supported by a review of the literature. PMID- 3053812 TI - Phagocytic and postphagocytic activities of bovine neutrophils for pure and mixed bacterial cultures. AB - A comparative study of phagocytosis and postphagocytic oxidative metabolic activity of bovine blood neutrophils incubated with pure and mixed cultures of Escherichia coli, Staphylococcus aureus, and Streptococcus agalactiae was preformed. Most neutrophils when incubated with mixed cultures showed preferential phagocytosis for one species and a smaller number phagocytized both species of microorganisms. Percent phagocytosis for E. coli in pure culture was similar to that of Strep. agalactiae in pure culture and higher than that for Staph. aureus in pure culture. Neutrophils incubated with mixed cultures of E. coli and Staph. aureus or E. coli and Strep. agalactiae showed greater than expected phagocytosis of each microorganisms alone and reduced phagocytosis of both microorganisms together. Postphagocytic oxidative metabolic activity of neutrophils, measured by percent nitroblue tetrazolium reduction, did not differ following phagocytosis of these three microorganisms in pure cultures. In comparison, a synergistic effect on nitroblue tetrazolium reductive activity was seen in mixed cultures as evidenced by higher percent nitroblue tetrazolium reduction following phagocytosis of both E. coli and Staph. aureus or E. coli and Strep. agalactiae. These observations indicate that the phagocytic and metabolic activities of neutrophils for bacteria in mixed cultures may not be identical to those in pure cultures. PMID- 3053815 TI - Udder health in the periparturient period. AB - The periparturient period is associated with rapid differentiation of secretory parenchyma, intense mammary growth, copious synthesis and secretion, and marked accumulation of colostrum and milk. Udder health during this time is an important factor associated with the production of maximum quantities of high quality milk. Intramammary infections that occur during the dry period can adversely affect udder health, resulting in decreased milk production, altered milk composition, and impaired mammary function. Bovine mammary glands are markedly susceptible to new infections during the periparturient period, especially prior to parturition. Many infections that occur at this time are associated with clinical mastitis during early lactation. Methods of controlling mastitis in the dry period have focused primarily on the use of antibiotics. However, antibiotic therapy at drying off does not appear to prevent new infections in the periparturient period. This is most likely due to lack of persistence of antibiotics. Furthermore, antibiotics used currently are less effective against environmental pathogens, in particular coliform bacteria, which can cause a high proportion of intramammary infections during the periparturient period. Methods of controlling bovine mastitis during the periparturient period is an important area that requires additional research. Procedures need to be developed that are effective against a variety of bacteria, including environmental mastitis pathogens, if additional control is to be achieved. PMID- 3053816 TI - A 2-year follow-up study of patient satisfaction with new complete dentures. PMID- 3053813 TI - Epidemiology of metabolic disorders in the periparturient dairy cow. AB - The descriptive epidemiology and risk factors were reviewed for six clinical disorders: milk fever, downer cow syndrome, hypomagnesemic tetany, udder edema, left displaced abomasum, and ketosis. Data were included also from preliminary analyses of a data set from approximately 61,000 Finnish Ayrshire cows. A web of postulated associations among the metabolic disorders and other risks factors (previous lactation diseases and milk yield, calf factors, certain dry period nutritional factors, dystocia, retained placenta, and metritis) was diagrammed. PMID- 3053814 TI - Reproductive disorders in the periparturient dairy cow. AB - Incidence, predisposing factors, and implications of various reproductive disorders (dystocia, twinning, stillbirth, retained placenta, cystic ovaries, anovulation, infections of the reproductive tract, metritis, and abnormal health status) are reviewed as to their inter-relationships and collective impact on reproductive performance, milk yield and predisposition to other diseases or disorders in the periparturient dairy cow. All reproductive disorders reviewed reviewed reduce reproductive performance either directly or indirectly. Concurrent milk yield was reduced marginally in a few studies as a consequence of twinning, retained placenta, cystic ovaries, metritis, or other uterine disorders, and in cows with an abnormal health status. There is strong evidence for associated losses in milk yield following surgical delivery of a stillborn calf. We conclude that most periparturient disorders occur as a complex, rather than as a single abnormality. Cows with one disorders are at increased risk for other disorders, including metabolic ones. In contrast, actual milk yield or potential for high production generally does not predispose cows to increased risk for any of the reproductive disorders. The literature suggests that prophylactic measures to prevent occurrence of the one disorder might decrease the risk and incidence of other related disorders, either directly or indirectly. PMID- 3053817 TI - Factors affecting the bond strength of composite resin to etched glass-ionomer cement. PMID- 3053818 TI - The effect of casting ring liners on the potential expansion of a gypsum-bonded investment. AB - Cellulose paper (used wet) and ceramic paper (used dry) are replacing asbestos paper as cushioning ring liners in dental casting. A study was made of the effects of all three of these materials on the setting and subsequent thermal expansion of a gypsum-bonded cristobalite casting investment (W/P = 0.40). Thermal expansion measurements were made on the same specimens that were produced during the setting expansion tests. Control specimens setting against a smooth dry surface showed a total expansion of 1.7%. Specimens setting against dry ceramic liners had similar total expansions, in the range 1.6 to 1.7%. Specimens setting against either of the wet lining materials showed an increased total expansion (in the range 2.2 to 2.3%), by virtue mainly of a large increase in setting expansion. In order for reproducible setting expansion results to be obtained with wet liners, it was necessary to control the amount of absorbed water carefully. Dry asbestos and dry cellulose liners gave higher expansions than pre-wetted ones, since they abstracted water from the mix, reducing its effective W/P ratio (giving a thicker mix), and then functioned as wet liners. These results suggest that, at least as far as potential investment expansion is concerned, wet cellulose liners have an effect similar to that of the traditional wet asbestos liners. Dry ceramic liners give a much lower investment expansion, and when these liners are used, an investment with an increased measured expansion could be an advantage. PMID- 3053819 TI - Influence of incisal length of ceramic and loading orientation on stress distribution in ceramic crowns. AB - For ceramic crowns, the recommended depth of tooth reduction from the incisal edge of anterior teeth is 1.5 mm to 2.0 mm. Although some prosthodontists have suggested that incisal heights of ceramic which exceed 2 mm are associated with dangerously high intra-oral stresses, this theory has not been verified. The objective of this study was to test the hypothesis that the stress distribution in ceramic crowns designed for a prepared maxillary central incisor which are subjected to applied loading is relatively insensitive to the incisal length of ceramic. Finite element stress analyses were performed on three crown designs loaded with a horizontal or vertical force of 200 N along the lingual surface near the incisal edge. Ceramic crowns for maxillary central incisors were modeled with incisal lengths of 1.0 mm (Case I), 1.9 mm (Case II), and 4.0 mm (Case III). Zinc phosphate cement with a film thickness of 30 micron was included for each case. Plane-stress finite element analyses indicated that tensile and compressive stresses which were induced in cement and ceramic due to a vertically applied load of 200 N were comparable in magnitude for all three cases within the gingival third of the crowns. For Cases I, II, and III, tensile stresses at the facial region were 6.7, 5.4, and 6.5 MPa, respectively, in cement and 46.2, 48.6, and 49.2 MPa, respectively, in the ceramic. The results of this study tend to support the hypothesis that the amount of tooth reduction (1 to 4 mm) in an incisogingival direction does not significantly influence the stress distribution in the crown or cement layer.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3053820 TI - A new ion-coating surface treatment of alloys for dental adhesive resins. AB - 4-META and new phosphate-methacrylate resins adhere strongly to dental alloys. However, for strengthening the water durability of the adhesive interface, the oxidation of the alloy surface is indispensable. A new oxidation method using ion sputtering was developed, and the effectiveness of this surface treatment on two dental alloys--a type IV gold alloy and Ni-Cr-Be alloy--was investigated. As an endurance test, thermocycling for a maximum of 100,000 cycles was adopted, and the tensile adhesive bond strength was then measured. Ion-coating the surface of the alloys resulted in strong bonds with adhesive resins, and after 100,000 thermocycles, a bond strength of above 20 MPa was maintained. PMID- 3053821 TI - Bone mineral content of the maxilla estimated by dual-photon absorptiometry after augmentation with bone or hydroxyapatite. AB - A new, precise, non-invasive method for measuring the bone mineral content (BMC) of the mandible in vivo by dual-photon absorptiometry has recently been introduced. The purpose of the present analysis was to examine the precision in vitro and in vivo and the accuracy in vivo for assessment of BMC in the maxilla. The precision was determined by repeated measurements in vitro on a cranium and in vivo on two test persons with and without a bone specimen fixed to a palatal plate. The accuracy in vivo was determined from the BMC measurements of the two test persons and of nine edentulous persons, scanned before and after augmentation of the maxillary alveolar ridge with hydroxyapatite. The analyses indicated that the precision for maxillary BMC assessments was high (0.9% in vitro and 2.0% in vivo) and the accuracy in vivo was 6.6%, corresponding to the accuracy in vitro for skeletal BMC measurements by dual-photon absorptiometry. The present method therefore seems to be well-suited for follow-up analyses of the BMC changes in the jaws after augmentation of the alveolar ridges with bone or hydroxyapatite. PMID- 3053823 TI - The dentist's role in access to dental care by Medicaid recipients. AB - This study uses a decision analytic approach to assess the dentist's role in access to care by Medicaid recipients. The question of whether a private dentist, when given the choice, will schedule a Medicaid or non-Medicaid patient is examined. The model considers factors frequently reported to influence dentist's decisions over whether to accept Medicaid recipients into their practices. Factors include reimbursement rates, probability of broken appointments, and likelihood of reimbursement. The model permits calculation of the expected benefits in dollars for comparable treatment of Medicaid and non-Medicaid patients. Under a variety of conditions, it is shown that the strategy to schedule a non-Medicaid patient dominates alternative strategies in which Medicaid recipients are scheduled. Policy implications of these results are discussed. PMID- 3053824 TI - The flashlamp-pumped 577-nm pulsed tunable dye laser: clinical efficacy and in vitro studies. AB - The 577-nm flashlamp-pumped tunable dye laser pulsed at 450 microseconds is rapidly becoming the treatment of choice for removal of port-wine stains and other vascular ectasias. In this study, we examined the mechanisms of vessel destruction by determining the effects of laser irradiation on three types of primary target cells in vitro: erythrocytes, endothelial cells, and fibroblasts. Clinical studies were also performed, addressing the efficacy of this laser in the treatment of port-wine stains of the head and neck, trunk, and extremities. In endothelial cell cultures, both [3H]thymidine (measuring cell proliferation) and [3H]leucine (measuring protein synthesis) incorporations were inhibited at energy levels of 5-12 J/cm2 (p less than 0.01). The laser energy in the range of 5-8.5 J/cm2 had no effect on cell viability. Erythrocytes as target cells for laser energy demonstrated rapid, dose-dependent lysis. Addition of erythrocytes into a co-culture with endothelial cells abolished the direct inhibitory effect noted in cultures when endothelial cells alone were present. The results of the latter experiment imply that erythrocytes are the primary target cell absorbing the laser energy at 577 nm. However, direct laser effects on endothelial cells may also contribute to the mechanisms of ablation of the vascular ectasias by the tunable dye laser at 577 nm. Clinical trials on 28 patients with port-wine stains of the face and neck using this laser demonstrated a 75% response rate with greater than 50% lightening of the lesions. Of 9 port-wine stain lesions on the trunk and extremities (on a total of 6 patients), 8 (89%) demonstrated significant (greater than 50%) fading of their lesions. Complications such as scarring or epidermal texture changes were minimal. PMID- 3053822 TI - The effects of sugars intake and frequency of ingestion on dental caries increment in a three-year longitudinal study. AB - A three-year longitudinal study was carried out with a group of children, initially aged 11-15, residing in non-fluoridated rural communities in south central Michigan. This report analyzes the relation between caries increment and consumption of sugars from all sources to see if accepted relationships have changed with the caries decline in the United States. There were 499 children who provided three or more 24-hour dietary recall interviews, and who received dental examinations at baseline and after three years. Caries increment averaged 2.91 DMFS over the three years, with 81% of new lesions on pit-and-fissure surfaces. Consumption of sugars from all sources averaged 156 g per day for males and 127 g per day for females, an average of 52 kg per person per year. Sugars constituted one-quarter of total caloric intake for both boys and girls, and the average number of eating occasions per day was 4.3. Children who consumed a higher proportion of their total energy intake as sugars had a higher increment of approximal caries, though there was little relation to pit-and-fissure caries. The average number of daily eating occasions was not related to caries increment, nor was the average number of sugary snacks (defined as foods with 15% or more of sugars) consumed between meals, but the average consumption of between-meal sugars was related to the approximal caries increment. When children were categorized by high caries increment compared with no caries increment, a tendency toward more frequent snacks was seen in the high-caries children.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3053825 TI - Neoplasms of the conchal bowl: treatment with Mohs micrographic surgery. AB - Thirty-four patients with tumors in the conchal bowl were treated with Mohs micrographic surgery. Twenty-six patients had basal cell carcinoma and 8 patients had squamous cell carcinoma. Thirty-three of 34 patients achieved a tumor-free margin during initial therapy. All patients remain free of recurrence and metastasis at an average follow-up period of 2.9 years. Thirty-three of 34 patients were allowed to heal by second intention. Only one patient required reconstruction to avoid stenosis of the external auditory canal. PMID- 3053826 TI - Allergic necrotizing eosinophilic granulomatosis. AB - Allergic necrotizing eosinophilic granulomatosis (ANEG) is a rare entity characterized by peripheral eosinophilia, hepatosplenomegaly, dyspnea, and lymphadenopathy. An unusual patient with ANEG is described in which the presenting feature was an extensive granulomatous lesion of the face. The patient's condition progressively deteriorated despite multiple therapies, and she finally succumbed to her disease. PMID- 3053828 TI - Haematoxylin stainable epidermal protein of the newborn rat. III. The change of the tissue reactivity to the antibody at various periods of growth. PMID- 3053827 TI - Comparison of the relative therapeutic efficacy of 7-methyl pyridopsoralen and 8 methoxypsoralen in photochemotherapy in psoriasis treatment. PMID- 3053830 TI - Characteristics of prolidase and prolinase in prolidase-deficient patients with some preliminary studies of their role in skin. PMID- 3053829 TI - Expression of transferrin receptor in normal human skin, psoriatic skin and various skin tumors. PMID- 3053831 TI - Tissue culture study of dermatofibrosarcoma protuberans. PMID- 3053832 TI - Experimental chronic dermatophytosis in human skin grafted to nude mouse: inoculation of Trichophytons and histopathological evaluation. PMID- 3053833 TI - Natural killer (NK) cell activity and NK-related cell surface markers in patients with atopic dermatitis. PMID- 3053834 TI - Monocyte-modulating activities in the sera of patients with granuloma annulare. PMID- 3053835 TI - Study of the skin of a new hairless rat mutant. PMID- 3053836 TI - Echographic studies of superficial lymphadenopathies. PMID- 3053838 TI - Colon carcinoma occurring two years after the development of SSSS in an adult. PMID- 3053837 TI - A case of dermatomyositis and Hashimoto's thyroiditis. PMID- 3053840 TI - [Use of the elements of an automated system for studying the action of substances on the operative memory of experimental animals]. PMID- 3053839 TI - [Changes in the composition of the human oral fluid in malignant neoplasms]. PMID- 3053842 TI - A retrospective survey of treatment and mortality in aspiration pneumonia. AB - A retrospective survey was conducted of all patients with severe aspiration pneumonitis requiring artificial ventilation in our Intensive Care Unit from 1982 1986 inclusive. Of 38 patients, 8 (21%) died. Five of these deaths were due to severe primary intracranial pathology, and occurred after complete or almost complete resolution of the pneumonitis. One death (2.5%) due to myocardial infarction was possibly related to aspiration, and 2 deaths (5%) were definitely related to aspiration. The 7.5% mortality related to aspiration is considerably lower than in previous clinical studies of severe aspiration pneumonia. There was only one death due to aspiration in patients under the age of 70. The mean arterial to alveolar oxygen tension ratio was 0.221, and the mean predicted mortality by apache II was 43%. Patients were managed with rapid intravascular volume restoration using crystalloid fluids, early ventilation, no steroids, and no immediate antibiotics. We conclude that with such management it is possible to achieve a low hospital mortality in severe aspiration pneumonia, particularly in young patients. PMID- 3053841 TI - The effect of nifedipine alone or combined with low dose acetylsalicyclic acid on endotoxin-induced pulmonary hypertension in the piglet. AB - Cardiovascular responses to the calcium antagonist nifedipine, alone and combined with low dose acetylsalicyclic acid (ASA), were evaluated in a piglet model of endotoxin-induced pulmonary hypertension. All animals were anesthetized, paralyzed and mechanically ventilated. Cardiac output (CO), pulmonary artery pressure (PAP), aortic blood pressure (SAP), pulmonary capillary wedge pressure (PCWP), right atrial pressure (RAPM) and arterial blood gases were measured before and after induction of pulmonary hypertension by E. coli endotoxin and after treatment. Results of treated groups were compared to a control group of piglets subjected to the same dose (0.15 micrograms/kg i.v.) of endotoxin. Control animals responded to a bolus injection of endotoxin within 15 min with an increase in mean PAP by 110%. Pulmonary vascular resistance (PVR) increased by 144%. Mean arterial pressure did not change significantly from baseline values. In animals treated with a single dose of 1 mg/kg ASA prior to endotoxin, the initial pulmonary response was not quantitatively different from control values, whereas ASA 20 mg/kg abolished the pulmonary vascular reaction. The increase of systemic vascular resistance (SVR) produced by endotoxin was aggravated by high dose ASA. In piglets treated with nifedipine (4 micrograms/kg/min) over 30 min after the application of endotoxin with and without additional infusion of nifedipine 60 min prior to endotoxin the PVR could be attenuated. The combination of nifedipine and low dose ASA showed synergistic effects compared to control. The increase of mean PAP was significantly reduced, the PVR remained in baseline range due to a marked elevation of cardiac output. PMID- 3053844 TI - Inspiratory effort and occlusion pressure in triggered mechanical ventilation. AB - We have studied eleven patients ventilated in the assisted mode during recovery from acute respiratory failure. We have measured the effort required to trigger the pressure demand valve for 3 different ventilators, and have measured the occlusion pressure as an index of neuromuscular inspiratory drive. We found a delay in the opening of the demand valve, as previously described by other authors. We also found a close correlation between the effort required to open the demand valve and the occlusion pressure. We conclude that the inspiratory effort required to open the demand valve, in the assist mode, is greater than the preset trigger level and that it is well correlated with the neuromuscular inspiratory drive. This inspiratory effort against the closed demand valve, allows the measurement of the occlusion pressure. PMID- 3053843 TI - Total inspiratory work with modern demand valve devices compared to continuous flow CPAP. AB - The inspiratory work exerted by an electromechanical lung model in drawing a 500 ml breath, was assessed by planimetry of pressure/volume loops for six commercial demand valve CPAP devices (Servo B and C from Siemens, EV-A and UV-2 from Drager, the Puritan Bennett 7200 and the Engstrom ERICA) and compared to the loading of a conventional high flow CPAP system. The effect of trigger sensitivity and inspiratory pressure support on inspiratory work was also investigated in some cases. The lung model allowed for calibrated changes in compliance and airway resistance. In the non-assisted CPAP mode, all machines required slightly larger amounts of inspiratory work than the continuous flow CPAP system. Most machines were comparable in performance but the ERICA and the Servo B required up to 22% more work than the continuous flow CPAP system and represented the maximal increase of total work due to any given machine. The greater part of total inspiratory work was due to lung compliance and airway resistance, factors external to the machines. Halving compliance doubled the work and exchanging a 7 for a 9 mm i.d. endotracheal tube in the circuit increased work by about 3% regardless of machine. Decreasing trigger sensitivity from 0 to 2 cm H2O for the Servo B increased work by up to 24%. Using 5 cm H2O of inspiratory pressure support decreased work for all machines up to 36% maximally.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3053845 TI - A high flow turbine CPAP system. AB - A continuous high flow CPAP system incorporating a turbine blower is described. The system achieves inspiratory flow rates of 150 l/min or more by means of reticulated gas flow and inspired oxygen fractions of 0.21-0.95. Positive airway pressure is provided by weighted disc valves and a modified aviation-type CPAP face mask provides electronic communication with the patient. The mobility of the system also enables its use as an intermittent physiotherapy aid. Work of breathing of the system, as assessed by total pressure fluctuations is at a minimum. PMID- 3053846 TI - Voices in distant camps: the gap between nursing research and nursing practice. PMID- 3053847 TI - Consultation: a review and analysis of the literature. PMID- 3053848 TI - Faculty practice: unifier of nursing education and nursing service? PMID- 3053850 TI - A treasury of dentistry. Toothpicks. PMID- 3053852 TI - The presidents. I. Lawrence Kerr 1979-1980. PMID- 3053851 TI - Research claim. PMID- 3053849 TI - Help wanted for SELECT program. PMID- 3053853 TI - AAWD: a voice for women in dentistry. PMID- 3053854 TI - Lucy Hobbs Taylor: the mixed blessing of being the first. PMID- 3053855 TI - Nonpainful, smooth-surfaced, bluish nodule on the upper lip. AB - A case of a vascular leiomyoma of the upper lip has been presented. A review of the literature was completed, and the findings reported. Leiomyomas are rare intraoral neoplasms. When affecting the mouth, they are most likely to occur on the tongue, palate, buccal tissues, and lips. Conservative local excision is the recommended treatment. PMID- 3053856 TI - Tooth-colored inlays and onlays. AB - Today's dental patient concerned about attractive restorations is looking for a dentist who incorporates the newest and best techniques into practice. This article discusses a newer concept that pleases both the patient and dentist: restorations that are bonded into acid-etched tooth preparations with composite resin cement--tooth-colored inlays and onlays. Basic questions asked by practitioners about these esthetic restorations are addressed; characteristics, performance expectations, comparisons with cast gold inlays and onlays, and comparisons with amalgam are presented, in addition to other relevant information for today's dental practitioner. PMID- 3053858 TI - The orthodontic appliance: esthetic considerations. AB - The importance of attractive dental and facial appearance is at an all-time high for the American consumer. Because of this emphasis on appearance, the esthetic impact of the orthodontic appliance is a matter of great concern to prospective patients. This article presents an overview of the esthetic features of currently available orthodontic appliances. PMID- 3053857 TI - Posterior composite resins. AB - A quarter of a century ago, composite resin was introduced and quickly outmoded silicate cement and acrylic resin as a restorative material. Although composite resin cannot replace amalgam, it has a strong foothold in dentistry, most notably for its superior color-matching ability. This article contrasts many properties of composite resin to amalgam, and takes a look at new developments in posterior esthetic materials. PMID- 3053859 TI - Esthetics and the edentulous patient. AB - Most individuals undergo some degree of psychological shock, depression, and loss of self-esteem after the loss of their teeth. Optimum success in the treatment of the edentulous patient can no longer be limited to the proper fit, function, speech, selection, and arrangement of teeth. In addition, today's dentists must use treatment strategies that include proper psychological management, caring, and recognition of the importance of the patient's perception of what is personally esthetic. This article addresses the esthetic considerations of this restorative challenge. PMID- 3053860 TI - Nonorganic failure to thrive in infancy: an update on nutrition, behavior, and growth. AB - Failure to thrive (FTT) has been defined in a number of ways, but most definitions include a weight less than the 5th percentile on the growth chart or a decreasing rate of weight gain. Nonorganic failure to thrive (NOFTT), i.e., FTT not due to organic disease, is the most common category of FTT in the United States and is associated with delayed growth and development and abnormal behaviors. Factors extrinsic to the infant are primarily responsible for NOFTT. That acute undernutrition may be a cause of the poor weight gain is suggested by anthropometric studies and by the observation that NOFTT infants often gain weight when food is supplied. Yet, decreased caloric intake has been documented in only a few infants, and not all infants immediately gain weight when given adequate calories. Current thinking attributes lack of weight gain in NOFTT to probably mixed interacting causes, including decreased nutrition, and abnormal hormonal mechanisms associated with abnormal behavior. That behavior and nutrition are related is recognized, but their interactions have not been adequately documented or explained. It is unknown whether behavior affects growth directly through nutrition or independently of nutrition. Until the cause or causes of poor growth and development in NOFTT are understood, permanently reversing the process will be difficult. This report reviews what is presently known about nutrition and growth in NOFTT. PMID- 3053861 TI - Nutritional dwarfing: growth, dieting, and fear of obesity. PMID- 3053863 TI - Child abuse and neglect: a diagnostic guide for the optometrist. AB - Since approximately 40% of all physically abused children have ocular complications, it is increasingly important for optometrists to be prepared for an encounter with a child patient who has been abused. This review of the literature was undertaken to develop a diagnostic guide for the optometrist in face of a growing incidence of child abuse. It describes different types of abuse and their physical, emotional and behavioral manifestations. A special emphasis has been placed on head trauma and associated ocular signs. A list of state child protective services agencies, and how to contact them, is included. PMID- 3053862 TI - Zinc in growth and development and spectrum of human zinc deficiency. AB - Growth retardation is seen in experimental animals as a result of severe dietary restriction of several essential trace elements. However, in humans, the effect of zinc deficiency is most pronounced. Growth failure and hypogonadism in males, related to a deficiency of zinc, have been recognized in many developing countries. A mild deficiency of zinc, affecting growth and development in children and adolescents, has been reported from developed countries as well. Zinc deficiency in humans may manifest as severe, moderate, or mild. The manifestations of severe zinc deficiency include bullous pustular dermatitis, alopecia, diarrhea, emotional disorder, weight loss, intercurrent infections due to cell-mediated immune dysfunctions, hypogonadism in males, neurosensory disorders, and problems with healing of ulcers. This condition can be fatal. A moderate level of zinc deficiency has been reported in a variety of conditions. Clinical manifestations include growth retardation and male hypogonadism in adolescence, rough skin, poor appetite, mental lethargy, delayed wound healing, cell-mediated immune dysfunctions, and abnormal neurosensory changes. A mild level of zinc deficiency may manifest with decreased serum testosterone level and oligospermia in males, decreased lean body mass, hyper-ammonemia, neurosensory changes, anergy, decreased serum thymulin activity, and decreased IL-2 activity. Although the clinical aspects of severe and moderate levels of zinc deficiency are well known, the recognition of mild levels of zinc deficiency has been difficult. Currently plasmas zinc appears to be the most widely used parameter for assessment of human zinc status, and it is known to be decreased in cases of severe and moderate deficiency of zinc.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3053864 TI - Essential factors in the care of elderly patients. AB - With the increase in life expectancy since the turn of the century, all optometrists must have a good understanding of essential factors in the care of elderly patients. Some of the most important aspects of the clinical approach to and understanding of the elderly patient are considered here. Characteristic differences in symptomatology, the altered doctor-patient relationship, polypharmacy and compliance issues, and altered environmental needs are also described. PMID- 3053865 TI - Assessment of hepatic function and damage in animal species. Animal Clinical Chemistry Association. AB - There are a wide variety of laboratory tests available to assess damage to and functional impairment of the liver, though the effectiveness of these tests varies greatly depending upon the type of damage and the animal species involved. Species differences in tissue localization, metabolism, specificity and sensitivity of parameters relating to the liver influence the choice of tests. Some tests can be applied usefully to most animal species while others may be highly specific in one species but show very low discriminatory potential in others. The tests available, and their use in veterinary and toxicological investigations have been reviewed in the light of current practice in the U.K. PMID- 3053866 TI - Dr Christopher Dresser and the Minton connection: botanist, artist, designer. PMID- 3053867 TI - Tolerance of the oral clonidine test in 75 pediatric patients. AB - We evaluated the tolerance and effectiveness of the oral clonidine test for GH in 75 children, 84% with hyposomia and 16% with other diseases. The test was well tolerated, since 97% of the examined children had no side effects with the exception of occasional drowsiness, pallor and myosis of short duration. Two of the children at the end of the test, had more severe symptoms 30 min after (deep asthenia, pallor and a further small blood pressure drop) which however, resolved after 4-6 h. No correlation was observed between the clinical picture and the drops in blood pressure and/or plasma cortisol in the children examined. We confirm the effectiveness of the clonidine test in the release of GH since in our study we observed no negative false subnormal responses. PMID- 3053868 TI - Exercise and longevity: studies on rats. PMID- 3053869 TI - Adrien Barr:ere and his caricatures of the medical faculty of the University of Paris:. "Surgeons and Gynecologists:. PMID- 3053872 TI - Discriminant function analysis to calculate a Crohn's activity group scale to predict future inactive or active disease. AB - A discriminant function analysis was performed on several demographic, anthropometric, clinical, and laboratory data of 685 observations performed over 12 months on 137 patients with Crohn's disease. A Crohn's activity group scale (CAGS) was calculated. The CAGS has two advantages over the usual Crohn's disease activity index: it is objective, but more important is the fact that the values, when calculated longitudinally, have predictive value. Thus, calculation of CAGS is useful for counseling purposes and may also be useful in the design of future trials assessing therapy for Crohn's disease by allowing prerandomization stratification of patients with high or low probability of future recurrences of symptomatic disease activity. PMID- 3053870 TI - Adrien Barr:ere and his caricatures of the medical faculty of the University of Paris: "The Professors:. PMID- 3053871 TI - The use of metronidazole (Flagyl) in Crohn's disease. AB - The efficacy of metronidazole in the treatment of Crohn's disease remains unproven. It has not been superior to a placebo in three small studies. Uncontrolled reports suggest it may be of benefit in selected patients who have not responded to previous medical or surgical therapy. Metronidazole therapy can lead to complete healing of perineal fistulas. Persistent concerns over side effects, carcinogenicity, and bacterial resistance suggest that drug should be prescribed with caution for long-term use. PMID- 3053874 TI - Metastatic septic endophthalmitis in pyogenic liver abscess. AB - In a consecutive series of 180 patients with pyogenic liver abscess, three patients (two men and one woman, between 46 and 75 years of age) had metastatic Klebsiella endophthalmitis. The incidence of metastatic endophthalmitis was 1.7% in patients with pyogenic liver abscess, 5.2% in patients with Klebsiella liver abscess, and 7.8% in patients with Klebsiella liver abscess having Klebsiella bacteremia. Despite aggressive therapeutic measures, the men permanently lost their vision and the woman eventually required an evisceration of her right eye. Delayed recognition and/or treatment as well as the nature of bacteria probably contributed to the tragic outcome. The findings suggest that a high index of suspicion is critical and a combined effort of the internist and ophthalmologist is mandatory. PMID- 3053875 TI - Fistula to the pancreatic duct from a peptic ulcer. Case report and literature review. AB - We report a case of pancreatic duodenal fistula as a complication of peptic ulcer disease. Complete healing of the ulcer and fistula was achieved with intensive medical therapy. No surgical intervention was required. The unique radiographic and endoscopic features of the case are presented and pertinent literature is reviewed. PMID- 3053873 TI - The role of colonoscopy in the diagnosis of tuberculosis. AB - The value of colonoscopy in the diagnosis of ileocolonic tuberculosis at Groote Schuur Hospital was assessed and the published world experience reviewed. During a 4-year period, abdominal tuberculosis was diagnosed in 94 patients, of whom 18 manifested involvement of the ileocecal area and/or colon. Colonoscopy, performed in 10 of these, provided diagnostic information in 7. Since the endoscopic appearance of diseased mucosa is nonspecific, adequate bacteriological and histological assessment of biopsied tissue is essential to differentiate tuberculosis from other disorders that may simulate it. We stress the importance of microscopy and culture for Mycobacterium tuberculosis. PMID- 3053876 TI - Severe bleeding from submucosal lipoma of the duodenum. AB - Two patients with a duodenal lipoma had severe gastrointestinal bleeding. Duodenal lipoma is relatively rare and an even rarer source of severe gastrointestinal bleeding. Endoscopy and routine radiological examination of the gastrointestinal tract may suggest the diagnosis, but computed tomography allows a definitive preoperative diagnosis. Surgery is indicated in order to avoid complications. PMID- 3053877 TI - Pancreatitis and duodenopancreatic reflux in Crohn's disease. Case report and review of the literature. AB - A 21-year-old woman with duodenal Crohn's disease developed pancreatitis many years after radiographic evidence of duodenopancreatic reflux. We review the 17 previously reported cases of non-drug-induced recurrent pancreatitis associated with Crohn's disease and discuss possible pathogenetic mechanisms. Pancreatitis should be considered in any Crohn's disease patient with filling of the pancreatic duct on barium study of the upper gastrointestinal tract. PMID- 3053878 TI - Caroli's disease of the left hepatic lobe associated with hepatic fibrosis. AB - We report a case of segmental Caroli's disease limited to the left hepatic lobe in a 32-year-old woman who presented with cholangitis. Histologic examination showed hepatic fibrosis also limited to the left lobe. PMID- 3053880 TI - An ultrasonic pearl. PMID- 3053879 TI - Saliva: its role in health and disease. AB - Saliva serves many functions in the upper digestive tract, with its roles in esophageal physiology, digestion, and gastric cytoprotection having been documented by many studies in the recent literature. Understanding the multiple functions of saliva and the consequences of its absence is of paramount importance to gastroenterologists. We present a clinically oriented review of the literature on the role of saliva in the upper gastrointestinal tract and review the many problems associated with lack of saliva. We also discuss the management of xerostomia. PMID- 3053881 TI - Roles of endogenous leukotrienes in damage caused by ethanol in isolated rat gastric cells. AB - The activity of prostaglandin (PG) and leukotriene (LT) synthesis and the role of endogenous LT in cellular resistance were assessed in isolated parietal and nonparietal cells of rats. Rat gastric cells were isolated and the fractions rich (F1) and poor (F2) in parietal cells were obtained. In F1, more PGE2, PGI2 (measured as the stable metabolite 6-keto-PGF1 alpha), and sulfidopeptide (SP) LTs and less thromboxane A2 (TXA2, measured as the stable metabolite TXB2) were synthesized than in F2. AA861, an inhibitor of 5-lipoxygenase, inhibited the synthesis of SP-LTs in both fractions in a dose-related way. AA861 alone at any concentrations used did not affect the viability of the cells, but prevented the cell damage caused by ethanol in both fractions, the effect of AA861 being inhibited by indomethacin. These results suggest that in rat stomach, PGs and LTs, except TX, are synthesized mainly in parietal cells rather than in nonparietal cells. Endogenous LT may play a crucial role in the development of cellular damage caused by ethanol independently of systemic and extracellular factors. The balance of PG and LT may be important for regulation of cellular resistance. PMID- 3053882 TI - Role of epidermal growth factor in gastroduodenal mucosal protection. AB - Epidermal growth factor (EGF) is a polypeptide produced in the submandibular glands, Brunner's glands, and the kidneys. The peptide is secreted in an exocrine fashion into saliva, duodenal juice, and urine. EGF stimulates cellular growth and differentiation and inhibits gastric acid secretion. Removal of the submandibular glands decreases the amount of EGF in saliva and gastric juice and subsequently the synthesis of DNA in the gastric mucosa is reduced as well as its resistance to bile-salt-induced gastric lesions. Intragastric instillation of EGF can prevent gastric ulcerations induced by aspirin as well as cysteamine in rats. EGF also accelerates the healing of chronic gastric ulcers induced by acetic acid. Cysteamine is a duodenal ulcerogen in rats. After cysteamine administration, the secretion of EGF from Brunner's glands decreases and the glands become depleted of mucus. Intraduodenal instillation of EGF can partly prevent formation of cysteamine-induced duodenal ulcers. Oral administration of EGF can accelerate healing of chronic duodenal ulcers in rats. The beneficial effect of EGF on healing of chronic gastroduodenal ulcers is probably due to the delayed effects of EGF such as stimulation of RNA and DNA synthesis. The protective effects of EGF are probably related to the early actions of the peptide such as activation of cell surface proteins and increased glycosaminoglycan synthesis. PMID- 3053883 TI - The role of smoking in peptic ulcer disease. AB - Cigarette smoking appears to be a risk factor for the development, maintenance, and recurrence of peptic ulcer disease. Smoking has an inconsistent effect on gastric acid secretion, but it does have other effects on upper gastrointestinal function that could contribute to the pathogenesis of peptic ulcer disease. These include (a) interference with the action of histamine-2 antagonists, (b) acceleration of gastric emptying of liquids, (c) promotion of duodenogastric reflux, (d) inhibition of pancreatic bicarbonate secretion, (e) reduction in mucosal blood flow, and (f) inhibition of mucosal prostaglandin production. Because these effects are related directly to the act of smoking and cessation of smoking is associated with the prompt recovery of the respective functions, smokers will benefit immediately by stopping or reducing cigarette consumption. PMID- 3053884 TI - Epithelial restitution in the gastrointestinal tract. AB - Injury to the gastrointestinal mucosa can be divided into two types: (a) deep injury involving extensive hemorrhage and large areas of tissue necrosis, and (b) superficial injury confined to the upper regions of the mucosa and not involving hemorrhagic lesions. Mucosal repair differs according to the severity of the damage. Repair of deep injury takes weeks because large regions of the mucosa must be replaced with new tissue, a process involving mitosis. Superficial injury is initially repaired rapidly over hours by epithelial restitution that does not involve mitosis but proceeds in the following sequence of events: First, the damaged surface epithelial cells are shed and form a layer that protects the restituting mucosa. Then the viable epithelial cells that remain attached to the mucosa at the margin of the wound become flattened and rapidly migrate over the denuded basal lamina. The superficial epithelium is re-established when migrating cells touch, form new tight junctions, and repolarize their organelles. This rapid protective mechanism, called restitution, has recently been documented in the stomach, duodenum, colon, and rectum. The cellular mechanisms, speed of migration, recovery of transmucosal electrical potential difference, and specific peculiarities of rapid epithelial restitution in the various regions of the gastrointestinal tract are reviewed here. PMID- 3053885 TI - The mucus barrier. Its role in gastroduodenal mucosal protection. AB - A layer of water-insoluble mucus gel has been shown to form a continuous cover over the gastroduodenal mucosal surfaces, of median thickness of 180 micron in stomach in humans. This adherent mucus is the first line in mucosal defence against the natural aggressors, acid and pepsin, in the lumen. Mucus gel provides a stable unstirred layer that supports surface neutralisation of acid by mucosal bicarbonate. Mucus gel is a diffusion barrier to pepsin in the lumen, preventing proteolysis of the underlying epithelial cells. There is, however, a dynamic balance between digestion by pepsin of the mucus layer at its luminal aspect and secretion of new mucus by the epithelium. There is evidence that, in peptic ulcer disease, the rate of peptic degradation of the mucus barrier is increased. Exogenous damaging agents such as ethanol and aspirin permeate the gel matrix of the mucus barrier, rapidly damaging the underlying epithelium. The subsequent reepithelialisation process is protected by a gelatinous coat over ten times thicker than the original adherent mucus layer. This gelatinous coat is primarily a fibrin-based gel with necrotic cells and mucus. PMID- 3053886 TI - Involvement of autonomic nervous system in gastric mucosal defense mechanism. AB - This article describes the histochemical, immunohistochemical, radioautographic, and ultrastructural localizations of aminergic and peptidergic nerves, neurotransmitter receptors, and their binding sites in the stomach wall. Cholinergic and vasoactive intestinal polypeptide (VIP)-ergic nerve fibers are distributed along the gastric microvasculature, within the myenteric and submucosal plexuses, and in the muscularis mucosae and circular muscle layer. In the mucosa, both nerve fibers evenly extend along the capillaries in association with the epithelial cells up to the mucosal surface. In particular, cholinergic nerves are proved to doubly innervate the mucosal capillaries and nonmuscular venules as well as the parietal cells. Adrenergic and neuropeptide Y (NPY) containing nerves are distributed primarily along the arterioles of the gastric microvasculature, within the myenteric plexuses, and in the circular muscle layer. These nerve fibers extend up to the basal portion of the mucosa in close association with small arterioles, capillaries, and epithelial cells. Some of the adrenergic nerve axons are coexistent with the cholinergic nerve axons within the Schwann cell. Histamine H1 receptors are widely located on the walls of arterioles, capillaries and venules, while H2 receptors are evident not only on the parietal cells but also on the walls of the collecting venules and surrounding capillaries in the mucosa. Dopamine D1 receptors are predominantly located on the smooth muscle cells of the arterioles near the muscularis mucosae, while D2 receptors are present on the walls of postcapillary venules and collecting venules. Functional coordination of both intramural peptidergic nerves as intrinsic origin and aminergic nerves as extrinsic origin is considered to be essential for maintaining the gastric mucosal defense mechanism against a variety of aggressive factors. PMID- 3053887 TI - Lactitol vs. lactulose in the treatment of chronic recurrent portal-systemic encephalopathy. AB - To compare the efficacy and patient acceptability of lactitol vs. lactulose in chronic recurrent portal-systemic encephalopathy (PSE), 25 cirrhotic patients with a history of repeated episodes of hepatic encephalopathy who required chronic administration of lactulose were included in a controlled cross-over clinical trial in which patients received, at random, lactitol (at an initial dosage of 10 g/6 h) or lactulose (15 ml/6 h, 66% w/v, containing 10 g of lactulose) during a 3 month period and then crossed-over to the alternative treatment for the following 3 months. Doses were adjusted to obtain two bowel movements per day. During the study period the daily protein intake was 40-60 g. Clinical and analytical data (including ammonia levels) were obtained, an EEG and the number connection test were performed and the PSE index was determined before treatment and monthly until the end of the treatment. No significant differences were found between the effects of lactitol and lactulose on the neurological and biological parameters, suggesting that the two treatments could be considered as equally effective. Lactitol was significantly better tolerated than lactulose (P = 0.02), the taste of which was assessed as being too sweet and provoking nausea. In conclusion, lactitol is a good alternative to lactulose for patients with chronic recurrent PSE, especially in those who do not tolerate the excessive sweetness of lactulose. PMID- 3053888 TI - Nadolol for prophylaxis of gastrointestinal bleeding in patients with cirrhosis. A randomized trial. AB - This controlled trial was designed to evaluate the prophylactic effect of nadolol on gastrointestinal bleeding in cirrhotic patients with large oesophageal varices who had never bled. Nadolol or placebo was given randomly to two groups of 53 patients. The percentage of patients free of gastrointestinal bleeding 1 year after inclusion in the study was 83 +/- 6% (mean +/- S.D.) in the nadolol group and 80 +/- 6% in the placebo group. In the nadolol and placebo groups, 40 and 47 patients, respectively, were compliant, i.e., took nadolol or placebo continuously. The percentage of patients who were free of bleeding 1 year after inclusion was 97 +/- 3% in the subgroup of compliant nadolol patients. This percentage was significantly higher than that of patients who were free of bleeding in the placebo group (P less than 0.03) as well as in the subgroup of compliant placebo patients (77 +/- 6%; P less than 0.02). We concluded that, although there was no overall significant effect of nadolol on the risk of bleeding in cirrhotic patients in good condition with large oesophageal varices, this study suggests that nadolol reduced the risk of bleeding in compliant patients. PMID- 3053889 TI - Causality assessment of drug-induced liver injury. Hepatology Working Group. AB - None of the various methods for drug reaction assessment has been adapted to the well-defined damage of a specific organ. There are disagreements between the parties involved in this issue: practitioners, and experts in drug surveillance either in regulatory agencies or in the pharmaceutical industry. Consensus Meetings were therefore organized in order to answer the three basic questions related to the diagnosis of drug-induced liver injury: (a) is the liver involved? (b) what chronological and clinical criteria suggest a drug-induced reaction? (c) can non-drug-related causes be excluded? On the basis of the method for drug reaction assessment used in France, we describe criteria adapted to the case of acute cytolytic hepatitis, which constitutes only one aspect of the task of the Hepatology Working Group. PMID- 3053890 TI - Cellular immune response to hepatitis B virus antigens. An overview. AB - It has been suggested that the cellular immune response to HBV antigens is responsible for hepatocellular injury in acute and chronic hepatitis B. However, definitive immunological studies have so far been hampered by the lack of appropriate model systems to study HBV antigen-specific T cells. The availability of highly purified and recombinant HBV antigens and of experimental techniques to maintain in continuous growth antigen-specific T cells derived not only from the peripheral blood but also from the liver should allow a better understanding of the fine immunopathogenetical mechanisms involved in viral clearance and liver damage. Whether some important biological characteristics of HBV antigens described in the mouse system, such as the high immunogenicity of the pre-S antigens and the capacity of the nucleocapsid of HBV to be a T cell-dependent and -independent antigen, are relevant to the immunopathogenesis of liver damage during natural HBV infection in man remains to be evaluated. PMID- 3053892 TI - Bibliography of the current world literature in hypertension. PMID- 3053891 TI - Splanchnic and hepatic renin metabolism in alcoholic cirrhosis. Lack of evidence of a splanchnic source of production or of significantly impaired hepatic clearance. AB - Cirrhosis is frequently associated with increased arterial plasma renin activity. This could be the result of increased renin production or diminished renin clearance. We measured plasma renin activity in simultaneous portal, hepatic vein, and femoral artery blood samples in 7 patients with clinically stable alcoholic cirrhosis to determine whether hepatic extraction of renin is reduced and whether, as has been suggested, there is a splanchnic source of plasma renin activity in this condition. Plasma renin activity (mean +/- S.E. in ng/ml/min) was similar in portal, arterial, and hepatic venous samples (portal: 8.0 +/- 3.7; arterial: 7.6 +/- 3.1; hepatic vein: 6.4 +/- 2.3). Hepatic extraction of plasma renin activity, calculated as [arterial-hepatic vein)/arterial) X 100, was 14 +/- 6%, not significantly different from reported normal values (26 +/- 3%, n = 46). The intrinsic hepatic clearance of plasma renin activity (235 +/- 89 ml/min) and the hepatic renin extraction rate (1.801 +/- 1.032 micrograms/min) were also similar to estimated normal values. The intrinsic clearance and extraction rates of renin correlated with arterial plasma renin activity (r = 0.93, P less than 0.01 and r = 0.79, P less than 0.05). These data indicate that in clinically stable patients with alcoholic cirrhosis: (1) hepatic renin clearance is not significantly impaired; and (2) there is not a splanchnic source of plasma renin activity. Therefore, increased peripheral plasma renin activity in this condition is due solely to increased renal renin production. PMID- 3053893 TI - Impaired baroreflex control of heart rate in high-renin renal hypertension. AB - The reflex responses of heart rate (HR) to bolus injection and infusion of phenylephrine and nitroglycerine were studied in rats with one-kidney, one clip (1-K, 1C) hypertension and hypertension produced by aortic ligation. Rats with mild 1-K, 1C hypertension (147 +/- 4 mmHg) and a normal renin-angiotensin system (RAS) activity had normal bradycardic and tachycardic responses, whereas rats with severe hypertension (208 +/- 3 mmHg) and over-active RAS showed impairment (70-80%) of the responses. After 12 days of aortic ligation rats with mild (146 +/- 2 mmHg) and severe (175 +/- 3 mmHg) hypertension exhibited overactivity of the RAS and marked impairment of the baroreflex control of HR. Normalization of RAS activity in rats with mild hypertension 60 days after aortic ligation, coincided with normalization of the bradycardic responses, whereas the abnormality persisted in those rats with severe hypertension in which hyperactivity of RAS also persisted. The response to 20-s carotid occlusion and the HR responses to electrical stimulation of the vagus nerve were normal in rats with renal hypertension, 12 days after aortic ligation. An ether inhalation test produced a marked reduction of 60% in the HR of control awake rats accompanied by a 46 +/- 5 mmHg increase in blood pressure. Reflex pathways other than those through the baroreceptors were largely responsible for the bradycardia since responses remained unchanged in rats with sino-aortic denervation (SAD) or when the pressure increase was prevented by phenoxybenzamine administration.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3053894 TI - Bibliography of the current world literature in hypertension. PMID- 3053895 TI - Pressure natriuresis and control of arterial pressure during chronic norepinephrine infusion. AB - The goal of this study was to quantitate changes in mean arterial pressure (MAP) and renal function during chronic increases in plasma levels of norepinephrine, and to determine the role of the renal pressure natriuresis mechanism in controlling sodium balance in norepinephrine hypertension. In six conscious dogs in which renal artery pressure (RAP) was allowed to increase during 7 days of norepinephrine infusion (0.2 micrograms/kg per min), sodium excretion (UNaV) rose from 66 +/- 3 to 112 +/- 15 mmol/day and MAP increased from 100 +/- 3 to 109 +/- 3 mmHg on the first day. On days 2-7, UNaV returned toward the control level while MAP averaged 108 +/- 2 mmHg. Glomerular filtration rate (GFR) and effective renal plasma flow (ERPF) did not change significantly, averaging 85.9 +/- 4.0 and 235 +/- 17 ml/min, respectively, during 7 days of norepinephrine, compared to controls of 84.1 +/- 3.9 and 252 +/- 20 ml/min. When RAP was servo-controlled for 7 days during norepinephrine infusion, the natriuresis was abolished; UNaV averaged 76 +/- 8 during control, 77 +/- 13 during the first day of norepinephrine and 65 +/- 4 mmol/day during 7 days of norepinephrine. GFR and ERPF did not change significantly during norepinephrine infusion with RAP held constant. MAP did not reach a plateau but continued to rise from 102 +/- 3 to 137 +/- 3 mmHg after 7 days of norepinephrine and servo-control of RAP.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3053896 TI - Hormone, calcium and blood pressure relationships in primary hyperparathyroidism. AB - The cause of hypertension in primary hyperparathyroidism and its response to corrective surgery remains a matter of controversy. We therefore studied blood pressure, vasoactive hormones and plasma calcium responses to parathyroidectomy in six hypertensive and two normotensive patients with primary hyperparathyroidism. Twenty-four-hour intra-arterial pressure recordings, together with hourly blood sampling for plasma renin activity (PRA), aldosterone, cortisol, catecholamines and calcium levels, were undertaken in each patient before surgery and were repeated under identical conditions 3-6 months after parathyroidectomy. Mean plasma calcium was 3.03 +/- 0.1 before, and 2.35 +/- 0.02 mmol/l after, parathyroidectomy. Changes in arterial pressure were small and variable in individual patients. Group mean arterial pressures before and after surgery were identical. Plasma cortisol and PRA were significantly higher in the hypercalcaemic state (P less than 0.01 and P less than 0.05, respectively) but there was no significant difference in plasma aldosterone or catecholamine levels. No correlations between changes in plasma calcium or parathyroid hormone levels and concomitant changes in plasma concentration of other hormones were observed. Our findings show that correction of primary hyperparathyroidism has no systematic effect on arterial pressure in a heterogeneous group, including some patients with probable background essential hypertension, when evaluated 3-6 months after surgery. Compared with values after corrective surgery, mean levels of PRA and cortisol-but not aldosterone or catecholamines--are elevated in patients with primary hyperparathyroidism. These findings are consistent with an inhibitory effect of raised ionic calcium concentration on the response of the adrenal glomerulosa to angiotensin and adrenocorticotrophic hormone. PMID- 3053897 TI - Lethal graft-vs-host disease induced by a class II MHC antigen only disparity is not mediated by cytotoxic T cells. AB - Treatment of C57BL/6J (B6) murine splenocytes with L-leucyl-L-leucine methyl ester (Leu-Leu-OMe) selectively removes NK cells, CTL precursors, and the capacity to cause lethal graft-vs-host disease (GVHD) in irradiated B6 X DBA/2 F1 mice. In contrast, alloantigen-induced L3T4(+) Th cell function has been shown to be relatively preserved after exposure to this agent. The present studies assessed the effects of Leu-Leu-OMe treatment of donor cells on induction of lethal GVHD in other murine strain combinations. When irradiated B6 X CBAF1 mice were infused with T and NK cell-depleted B6 bone marrow cells and 3 to 30 X 10(6) B6 spleen cells, uniformly lethal GVHD was observed. However, B6 X CBAF1 recipients of T and NK-depleted B6 bone marrow cells and similar numbers of Leu Leu-OMe-treated B6 spleen cells demonstrated 90 to 100% long term survival. In contrast, Leu-Leu-OMe treatment of B6 donor cells had no beneficial effect on mortality rates in irradiated (B6 X B6-C-H-2bm12)F1 (B6 X bm12F1) recipients. When B6 spleen cells were stimulated in vivo or in vitro with either B6 X CBAF1 or B6 X bm12F1 stimulator cells, the capacity to generate alloantigen-specific CTL was abolished comparably by Leu-Leu-OMe treatment. Thus, the dramatic difference between the effects of Leu-Leu-OMe treatment of B6 spleen cells on the course of GVHD in B6 x CBAF1 and class II MHC only disparate B6 x bm12F1 recipients could not be explained by unique resistance of bm12-specific CTL precursors to Leu-Leu-OMe. These findings indicate that T cell effector mechanisms distinct from classic cell-mediated cytotoxicity are sufficient to generate lethal GVHD in class II MHC only disparate B6----B6 X bm12F1 mice. PMID- 3053898 TI - Effects of granulocyte-macrophage colony-stimulating factor and colony stimulating factor-1 on the proliferation and differentiation of murine alveolar macrophages. AB - Murine alveolar macrophages (AM) were shown to have proliferative ability and to form colonies in vitro. The factors in lung-conditioned medium (CM) and L929-CM which stimulate the proliferation of AM were considered to be granulocyte macrophage colony-stimulating factor (GM-CSF) and CSF-1, respectively, because recombinant murine (rm)GM-CSF and recombinant human (rh)CSF-1 could replace the activities of lung-CM and L929-CM, respectively. The phenotype of the cells in the colonies formed by AM incubated with rmGM-CSF or lung-CM was AM-like; more than 90% of the cells were stained by anti-asialo GM1 but not by FITC-LPS, and had AM-like morphology. Expression of Mac-1 Ag determined by M1/70HL in these cells as well as original AM was low. However, the phenotype of the cells in the colonies formed by AM incubated with rhCSF-1 or L929-CM was peritoneal macrophage (PM)-like; more than 90% of the cells were stained by FITC-LPS and M1/70HL, but not by anti-asialo GM1, and showed PM-like morphology. The cells in the colonies formed by AM incubated with rmGMCSF changed their phenotype after treatment with rhCSF-1; the percentage of cells stained by anti-asialo GM1 decreased, and that of cells stained by FITC-LPS increased. The cells in the colonies formed by AM incubated with rhCSF-1 never changed their phenotype after incubation with rmGM CSF. In contrast to AM, more than 90% of the cells in all colonies formed by PM incubated with either rmGM-CSF, rhCSF-1, lung-CM, or L929-CM were stained by FITC LPS but not by anti-asialo GM1. These results show that although AM and PM can proliferate, AM, in contrast to PM, are bipotential cells that can differentiate into two types of macrophages responding to distinct types of CSF, and that one of the molecular mechanisms controlling macrophage heterogeneity may be based on the type of CSF produced at distinct tissues. PMID- 3053899 TI - The effect of human recombinant macrophage colony-stimulating factor (CSF-1) on the murine mononuclear phagocyte system in vivo. AB - Human recombinant macrophage CSF (CSF-1) was administered i.v. to mice. After four daily injections there was a dose-dependent increase in the responsiveness of bone marrow cells from the treated animals to CSF-1 in vitro. At the highest dose tested (20,000 U/day) there was a selective 10-fold increase in the circulating population of mature monocytes. CSF-1 treatment also increased the macrophage content of the liver and peritoneal cavity and caused splenomegaly. The macrophages isolated from the peritoneum of CSF-1-treated animals were larger and expressed higher levels of the macrophage-specific F4/80 Ag. These data demonstrate that CSF-1 can act as a circulating regulator of the mononuclear phagocyte system. PMID- 3053900 TI - Group B streptococci inhibit the chemotactic activity of the fifth component of complement. AB - Infection with group B streptococci (GBS) is associated with a poor acute inflammatory response in which neutrophils fail to localize at the site of invasion. In the present studies, we have examined the effects of group B streptococci on C-derived chemotactic activity in human serum. Fresh human serum was activated to form C5a and C5adesarg by incubation with zymosan. The activated serum was then incubated with group B organisms, centrifuged, and the supernatants tested for chemotactic activity for human polymorphonuclear leukocytes. Group B organisms caused a dose-dependent decrease in C-dependent chemotactic activity. The degree of inhibition was profound with 1 X 10(9) bacteria/ml (10% of control). Experiments indicated that significant chemotactic factor inactivation occurred within 2 min of exposure to GBS organisms, while maximal inhibition occurred after 30 min incubation. A number of different strains of GBS of types I, II, and III possessed inhibitory activity. In contrast, group D streptococci, Staphylococcus aureus, Escherichia coli and Klebsiella pneumoniae failed to inhibit the C-derived chemotactic activity in human serum. Group A streptococci that were M protein positive also inactivated C dependent chemotactic activity in serum, as previously reported. The inhibitory activity of the GBS strains could be abolished by heat or trypsin treatment but not by neuraminidase, pronase, or pepsin. C5a levels in zymosan-activated serum as measured by RIA were not decreased after incubation with an inhibitory strain suggesting that absorption was not involved. SDS-PAGE analysis revealed that group B streptococci degrade the C5a molecule, increasing its electrophoretic mobility by removing a fragment with a m.w. of approximately 650 Da. Thus, one of the reasons for the poor inflammatory response at the site of GBS infection may reside in the ability of these pathogens to inactivate C-derived inflammatory mediators. The GBS C5a-ase activity probably serves as an additional virulence factor for these organisms contributing to the poor inflammatory response characteristic of group B streptococcal infection. PMID- 3053901 TI - Enhancement of human monocyte tumoricidal activity by recombinant M-CSF. AB - Activated monocytes are an important component of immunologic defense against neoplastic disease. A variety of agents capable of inducing tumoricidal activity have been described, including bacterial LPS, IFN-gamma, IL-1, IL-2, TNF, and GM CSF. We now show that pretreatment of monocytes with recombinant human macrophage specific colony stimulating factor (M-CSF) augments the tumoricidal activity of human peripheral blood monocytes induced by other activating agents. Monocytes were preincubated for three days with M-CSF at 10(3) U/ml, washed, and treated for an additional two days with secondary activators. Tumoricidal activity was measured in a 6-h 51Cr-release assay using NK-resistant WEHI 164 cells that had been treated with actinomycin D. Pretreatment of monocytes with M-CSF significantly increased tumoricidal activity induced by LPS, IFN gamma, LPS plus IFN gamma, and LPS plus PMA. Pretreatment with IL-1, IL-2, IL-3, IL-4, or GM-CSF was not as effective as M-CSF in increasing tumoricidal activity. Enhanced tumoricidal activity was directly correlated to the increased TNF production resulting from M-CSF pretreatment. TNF antiserum completely blocked tumoricidal activity, demonstrating that TNF was responsible for the M-CSF-mediated increase in tumor cell lysis. M-CSF pretreatment also enhanced non-TNF mediated tumoricidal activity by monocytes, as seen by increased killing of the TNF resistant target P815. This study demonstrated that in addition to the role of M CSF in the proliferation and differentiation of monocyte/macrophage precursors, M CSF also augments an effector function of mature blood monocytes. PMID- 3053903 TI - Biochemical characterization of membrane cofactor protein of the complement system. AB - Membrane cofactor protein (MCP; formerly termed glycoprotein 45-70 to indicate its Mr) of complement is a widely distributed iC3/C3b binding protein with co factor activity. On human mononuclear cells and cell lines and platelets, MCP is a doublet. The two forms differ in Mr by approximately 5 k and the upper species is predominant in most individuals. To further characterize these two forms, limited proteolytic digestions were performed. Of the four peptides produced, three have identical Mr indicating that the molecules are similar proteins. Both forms also have acidic isoelectric points and shift to a less acidic isoelectric point after treatment with neuraminidase. Glycosidase digestions indicate that both species contain N- and O-linked oligosaccharides but that the quantity of sialic acid is greater on the larger one. Pulse-chase experiments demonstrate approximately equal quantities of two precursor forms with Mr of 41 and 43 k. These two precursors possess N-linked high-mannose type of oligosaccharides and chase into the mature molecules which have complex sugars. The smaller precursor chases at a slower rate, possibly accounting for the reduced quantity of the smaller form of the mature form of MCP. These experiments indicate that the two forms of MCP are structurally similar and are derived from two distinct precursors. They also suggest that variations in the rate of processing of two intracellular precursors may account for the different quantities of the mature forms of this membrane protein. PMID- 3053902 TI - Synergistic effects of purified recombinant human and murine B cell growth factor 1/IL-4 on colony formation in vitro by hematopoietic progenitor cells. Multiple actions. AB - Purified recombinant human B cell growth factor-1/IL-4 was evaluated, alone and in combination, with purified preparations of recombinant human (rhu) CSF or erythropoietin (Epo) for effects on colony formation by human bone marrow CFU-GM progenitor cells (GM) and burst forming unit-E progenitor cells. rhu IL-4 synergized with rhu G-CSF to enhance granulocyte colony formation, but had no effect on CFU-GM colony formation stimulated by rhu GM-CSF, rhu IL-3, or rhu CSF 1. Rhu IL-4 synergized with Epo to enhance BFU-E colony formation equal to that of Epo plus either rhu IL-3, rhu GM-CSF, or rhu G-CSF. Removal of adherent cells and T lymphocytes did not influence the synergistic activities of rhu IL-4. Rmu IL-4, synergized with rhu G-CSF, but not with rmu GM-CSF, rmu IL-3, or natural mu CSF-1, to enhance CFU-GM (mainly granulocyte) colony numbers by a greater than 90% pure preparation of murine CFU-GM. Also, rhu IL-4 at low concentrations enhanced release of CSF and at higher concentrations the release also of suppressor molecules from human monocytes and PHA-stimulated human T lymphocytes. Use of specific CSF antibodies suggested that rhu IL-4 was enhancing the release of G-CSF and CSF-1 from monocytes and the release of GM-CSF and possibly G-CSF from PHA-stimulated T lymphocytes. Use of antibodies for TNF-alpha, IFN-gamma, or TNF-beta as well as measurement of TNF and IFN titers suggested that the suppressor molecule(s) released from monocytes were acting with TNF-alpha and those released from PHA-stimulated T lymphocytes were acting with IFN-gamma. These results implicate B cell growth factor-1/IL-4 as a synergistic activity for hematopoietic progenitors and suggest that the actions can be on both progenitor and accessory cells. PMID- 3053904 TI - HLA-B27 in inbred and non-inbred transgenic mice. Cell surface expression and recognition as an alloantigen in the absence of human beta 2-microglobulin. AB - A gene encoding the H chain of the human class I MHC Ag HLA-B27 was introduced into the germ lines of inbred C57BL/6 (B6) and non-inbred (B6 X SJL/J) F2 mice. By immunofluorescence and flow cytometry, the HLA-B27 gene product was expressed on lymphoid cells at levels comparable to the endogenous H-2b and H-2s class I MHC molecules. In both primary and secondary MLC between responder spleen cells from non-transgenic (B6 X SJL/J) F1 mice and transgenic stimulator cells, CTL were generated that specifically lysed mouse L cell (H-2k) or human B cell targets expressing HLA-B27, and this lysis thus appeared largely unrestricted by H-2. These results indicate that transgenic mice express a functional HLA-B27 gene product on cell surfaces in the absence of the human beta 2-microglobulin gene. These transgenic mice promise to be a valuable resource in the investigation of the unique role of HLA-B27 in inflammatory human disease. PMID- 3053905 TI - Tumor necrosis factor induction by Candida albicans from human natural killer cells and monocytes. AB - Other investigators have previously reported that TNF has been induced from macrophages by bacteria and, more recently, from NK cells by certain tumor cells. Sendai virus has also been reported to induce TNF from macrophages. We report here that an opportunistic fungi, Candida albicans, can also induce TNF, not only from human monocytes, but also from Percoll-fractionated large granular lymphocytes (LGL) which mediate NK function. Incubation of monocytes of LGL with C. albicans for 8 h was sufficient for detection of TNF release and peak induction was observed at 24 h. Induction of TNF from LGL did not require the participation of monocytes or T cells because treatment of the LGL with CD14 or CD15 to eliminate contaminating monocytes and CD3, CD4, or CD8 to eliminate contaminating T cells did not decrease the level of TNF produced from the treated LGL. Small T cells recovered from the denser fractions of the Percoll gradient had no ability to produce TNF, even when 10% monocytes were added to the T cells to provide accessory function. The phenotype of the TNF-producing LGL was CD2+, CD11+, CD16+, NKH1+, LEU7-. The TNF produced by both monocytes and LGL was neutralized by specific monoclonal and polyclonal anti-TNF but not by monoclonal antilymphotoxin. These results indicate that TNF production is a normal response of monocytes and LGL to stimulation by fungi such as C. albicans and that the release of TNF may be related to its ability to activate effector function to control Candida growth, which we have shown earlier for neutrophils with TNF. PMID- 3053906 TI - Use of regression analysis and the complement-dependent cytotoxicity typing assay for predicting lymphoid chimerism. AB - The complement-dependent cytotoxicity typing assay was studied for its accuracy in determining the presence of donor lymphocytes within standard chimeric donor/host cell combinations. Regression analysis of the data was utilized to evaluate the chimera assay. Excellent coefficients of determination (r2 greater than 0.90) were obtained for all standard curves, and a significant (P less than 0.001) linear relationship was established between percent cytotoxicity (dependent variable) and level of donor target cell chimerism (independent variable) for each regression equation. A highly significant (P less than 0.001) linear function was also established between percent cytotoxicity and concentration of donor target bone marrow cells. Regression coefficients (slopes) approached, but did not show complete unity (range; b = 0.86-0.95). Therefore, levels of cytotoxicity were not directly equivalent to levels of donor cell chimerism. A more accurate assessment of donor lymphoid chimerism would be provided by regression analysis of standard donor/host cell independent variables and inverse prediction. Significant estimates of peripheral donor lymphoid chimerism in putative mixed chimeric recipients were successfully made by this technique. PMID- 3053908 TI - Detection and quantitation of anti-neutrophil cytoplasm antibodies in Wegener's granulomatosis by ELISA using affinity-purified antigen. AB - Autoantibodies directed against cytoplasmic components of neutrophil granulocytes and monocytes (ACPA) have previously been described as a disease-specific marker for Wegener's granulomatosis (WG). We have developed an ELISA for determining and quantifying ACPA using an affinity-purified antigen preparation. The antigen was purified from supernatants of human neutrophils stimulated with phorbol ester to induce degranulation, by means of affinity chromatography with naturally occurring human autoantibodies. The established ELISA was sufficiently specific and sensitive, and the ACPA concentrations obtained with it correlated significantly with the ACPA titres determined by an indirect immunofluorescence technique (Spearman's rank correlation coefficient = 0.85, P less than 0.001, n = 105 WG patients). This ELISA provides precise ACPA quantitation and should prove valuable for monitoring disease activity in WG. PMID- 3053907 TI - Enzyme immunoassay analysis of antibody specificities present in the circulating immune complexes of selected pathological sera. AB - Using immobilized anti-C3 antibody and an enzyme immunoassay, sera from 26 patients (eight with systemic lupus erythematosus (SLE), four with Hashimoto's thyroiditis, eight haemophiliacs and six with post-hepatitis cirrhosis) containing high levels of circulating immune complexes (IC) were selected. The IC were precipitated with 2.5% polyethylene glycol, washed, treated with acid buffer, neutralized and tested using an enzyme immunoassay in parallel with the original sera for antibody activity against a panel of antigens: human myosin and thyroglobulin, mouse actin and tubulin, calf thymus DNA and trinitrophenyl coupled to bovine serum albumin (TNP/BSA). It was found that all the isolated IC may contain IgG, IgA and IgM antibodies reacting with actin tubulin and TNP/BSA and also, depending upon the disease, antibodies reacting with some of the other antigens of the panel. By comparison to the antibodies present in the original sera, higher titers of antibodies were found in the isolated IC while some antibody specificities not detected in a given serum were occasionally noted in the isolated IC. The antibodies present in the IC seem to possess characteristics similar to those of polyreactive human natural autoantibodies. It is concluded that natural autoantibodies participate actively in the formation of IC found in pathological sera. PMID- 3053909 TI - Assessment of anti-HLA antibodies in sera being tested for platelet reactivity by a platelet lymphocyte immunofluorescence test (PLIFT). AB - An indirect immunofluorescence test using a platelet mononuclear cell suspension is described. The test is a simple and sensitive method to assess the contamination by anti-HLA antibodies of sera being tested for platelet reactivity and eliminates the need to support two independent test systems such as lymphocytotoxicity and immunofluorescence testing. The test could be of value in routine platelet serology testing and platelet crossmatching. PMID- 3053910 TI - Cloning of myelomas and hybridomas in fibrin clots. AB - Myelomas and hybridomas were observed to proliferate normally when trapped in a fibrin clot. The fibrin clot was obtained by including fibrinogen in the cell culture medium and thrombin in the plastic dish. A clot formed within seconds and cloning efficiency was around 100%. This technique has all the advantages of semi solid medium cloning but avoids toxicity to the cells and the exposure to high temperature associated with the soft-agar cloning technique. PMID- 3053911 TI - Evaluation of monoclonality of cell lines from sequential dilution assays. Part III. PMID- 3053912 TI - Septic arthritis due to Cryptococcus neoformans. AB - Cryptococcal arthritis is rare. We report two cases, one of infection of the hip joint in a 56-year-old woman, the other of arthritis of the elbows and knees in a 4-year-old boy. Both patients were treated successfully with a combination of surgical drainage and antifungal therapy. The 15 previously published cases are reviewed. Immunodeficiency is noted in most reported cases commonly associated with the use of corticosteroids. The joint most often involved is the knee. Amphotericin B and 5-fluorocytosine have been used with success for treating this conditions. Ketoconoazole may have a role in the therapy of cryptococcal arthritis although information on the use of this agent is sparse. PMID- 3053914 TI - Fatal septicaemia with group A streptococcal infection in previously healthy subjects. PMID- 3053913 TI - Cerebral malaria from West Africa: chloroquine-resistant Plasmodium falciparum? PMID- 3053915 TI - Dysentery and Streptococcus pyogenes. PMID- 3053916 TI - Is pneumococcal infection a preventable disease? PMID- 3053917 TI - The structure, pathogenicity and genetics of Haemophilus ducreyi. PMID- 3053919 TI - Recognition and laboratory characteristics of an atypical oocyst of Cryptosporidium. AB - Feces from some patients with clinically unremarkable cryptosporidiosis contained an unusual variety of oocyst not previously recognized. These atypical oocysts were shown by electron microscopy to have a distinctive three-layered outer coat and, by immunofluorescence with a monoclonal antiserum, to lack an antigen present on the surface of typical oocysts. In contrast to typical oocysts, the atypical variety is very fragile and quickly collapses when suspended in solutions of high osmotic pressure or in lipid solvents. Atypical oocysts cannot be stained by methods used to stain typical oocysts, but their appearance in sucrose-phenol is characteristic. Their stability in this solution, though much less than that of typical oocysts, is sufficient for them to be recognized and for cases to be diagnosed by microscopy. Patients who excreted atypical oocysts never excreted the typical variety. General findings in patients who excreted atypical oocysts were no different from those who excreted typical oocysts. PMID- 3053918 TI - Immune response to acute diarrhea seen as circulating antibody-secreting cells. AB - We studied the immune response to acute diarrhea by examining antibody-secreting cells among peripheral blood lymphocytes, which are believed to be derived from the intestinal mucosa and to be on their way back there. In 23 of 24 patients, a dramatic increase in the total number of cells actively secreting immunoglobulins was detected one week after onset of diarrhea, and most of the cells were secreting IgA. Cells secreting antibody specific to the pathogen (Campylobacter jejuni or Salmonella spp.) also appeared at this time but accounted for only a part of the total response. The data suggest that diarrhea induces a vigorous, apparently polyclonal response, including antibodies to normal intestinal flora. The response to the infective agent was outstanding and suggests that this method can be used to identify the causative agent of an infection. PMID- 3053920 TI - Diagnosis and successful treatment of fusariosis in the compromised host. AB - We present six cases of fusariosis caused by Fusarium solani that were diagnosed over a three-year period in 166 adult patients undergoing chemotherapy for acute leukemia. Necrotic skin lesions were evident in four patients, fungemia in three, and a deep cellulitis around a great toe nail at the time of a febrile illness in two. The mean minimal inhibitory concentration (MIC) of amphotericin B was 3.3 micrograms/mL and of miconazole, 5.3 micrograms/mL; all isolates were resistant to 5-fluorocytosine. All patients received amphotericin B (1.0-1.5 mg/kg per d) plus 5-fluorocytosine. In contrast to results found in the literature, five patients had resolution of their infections, and the one patient who died had necropsy evidence of disseminated fusariosis. Review of our cases and of the literature did not reveal a definitive source for the organism or its portal of entry. Fusarium spp. must be recognized as opportunistic pathogens that cause a potentially fatal infection in compromised patients. PMID- 3053921 TI - Functional versus phenotypic analysis of T cells in subjects seropositive for the human immunodeficiency virus: a prospective study of in vitro responses to Cryptococcus neoformans. AB - We performed a prospective study of 50 subjects at high risk for human immunodeficiency virus (HIV) infection to determine if assays of antigen-specific T cell function provide an earlier indication of future progression to AIDS or a better assessment of immune function than do current methods of evaluation. We measured in vitro T cell responses to Cryptococcus neoformans and tetanus toxoid, response to mitogens, HIV p24 antigenemia, and clinical parameters. Progression to AIDS was significantly associated with loss of T cell response to cryptococci (P = .015), HIV antigenemia (P = .001), and low CD4+ cell numbers (P = .001). Most importantly, we found that loss of antigen-specific responses to cryptococci and tetanus can occur before changes in CD4 cell number. Abnormal response to mitogens and marked depletion of CD4+ cells were late signs of progressive HIV infection. Measurement of antigen-specific T cell function may be useful for assessing the efficacy of antiviral therapy in HIV infection before onset of symptoms. PMID- 3053922 TI - The AIDS dementia complex. PMID- 3053923 TI - Acute lower-respiratory-tract infection associated with chlamydial TWAR antibody in Filipino children. PMID- 3053924 TI - Comparison of serum and fecal antibody responses of patients with naturally acquired Shigella sonnei infection. PMID- 3053925 TI - Prophylactic treatment of vivax and falciparum malaria with low-dose doxycycline. PMID- 3053926 TI - Characterization of the human antibody response to an Escherichia coli O111:B4 (J5) vaccine. PMID- 3053927 TI - Effects of in vivo administration of recombinant human granulocyte-macrophage colony-stimulating factor on human neutrophil chemotaxis and oxygen metabolism. PMID- 3053928 TI - Karel Raska, 1909-1987. A tribute. PMID- 3053929 TI - Characterization of the components in circulating immune complexes from infants with congenital syphilis. AB - Immunoglobulin class-specific Clq-solid-phase assays were used to detect circulating immune complexes in the sera of infants with congenital syphilis. Elevated levels of IgM complexes were present in the sera of all infants with overt disease and in two of 14 asymptomatic infants considered to be "at risk." The sera of infants born to normal, serofast, and biologic false-positive mothers did not contain immune complexes. Immunoblotting techniques revealed that complexes isolated from the sera of the infected infants contained endogenous host antigens, as well as a limited number of treponemal polypeptides. Consistent with earlier findings examining purified immune complexes from adults with secondary syphilis and from infected animals, an 83-kilodalton Treponema pallidum antigen was present in all of the isolated complexes from infants with congenital syphilis. Our findings emphasize the fact that current serological and clinical measures of infection are inadequate and that certain "at risk" infants should be treated despite normal cerebrospinal findings and the absence of clinical manifestations. PMID- 3053930 TI - Two novel antigens associated with group B streptococci identified by a rapid two stage radioimmunoassay. AB - A two-stage radioimmunoassay (RIA) has been developed that identifies type specific antigens on the surfaces of group B streptococci. In addition to detecting the type-specific carbohydrate antigens Ia, Ib, II, and III, the RIA identified four unique antigens reactive with the Centers for Disease Control's c protein typing antiserum. Analysis of hot-acid extracts by immunoelectrophoresis confirmed that two of these reactivities corresponded to the reported alpha and beta antigens of the c-protein marker. In addition, the identification of two heretofore unidentified antigens, gamma and delta, by the two-stage RIA is detailed in this report. The assay uses intact bacteria and does not require hot acid extraction, a procedure thus enabling detection of acid-labile antigens in their native unmodified form. The semiquantitative RIA requires less typing reagent than does precipitin testing and is more objective, reproducible, and rapid. The assay described here could be applied to the detection of any cell surface antigen for which a monospecific antiserum is available. PMID- 3053931 TI - Isotype distribution and bactericidal activity of antibodies after immunization with Haemophilus influenzae type b vaccines at 18-24 months of age. AB - The serum antibody response to Haemophilus influenzae type b capsular polysaccharide or its protein conjugate vaccine (PRP and PRP-D, respectively) was studied in 28 children initially immunized at the age of 24 mo with either vaccine and in 10 children immunized for the third time with PRP-D at the age of 18 mo. The methods used were isotype-resolving enzyme immunoassay, Farr-type radioimmunoassay, and the in vitro bactericidal activity (BCA) test. Immunization with PRP evoked a higher proportion of IgA antibodies than did either the first or third dose of PRP-D, whereas the latter vaccine evoked a somewhat higher IgG response, but the differences were not statistically significant. In all groups the IgG antibody responses were predominantly IgG1, with the mean proportions being 82.2%, 84.2%, and 65.9% in the PRP, first-dose PRP-D, and third-dose PRP-D groups, respectively. Postimmunization antibodies were functionally active in the BCA test. PMID- 3053932 TI - Vascular endothelial cells and hematopoietic regulation. AB - Vascular endothelial cells elaborate growth factors which support the proliferation and differentiation of hematopoietic progenitor cells. Both the regulation and potential biological significance of growth factor production by endothelial cells are reviewed. PMID- 3053934 TI - [Relationships between extravascular lung water volume using thermal-dye dilution technique and pulmonary functions in patients undergoing pneumonectomy]. PMID- 3053933 TI - Improvement of human myeloma stem cell growth in a liquid culture system supplemented with phytohemagglutinin. AB - A highly efficient cloning system for in vitro myeloma colony growth could be valuable for screening antineoplastic agents in resistant patients and for testing the effects of purging methods in the context of autologous bone marrow transplantation. In this paper we report the results of experiments intended to improve the myeloma cloning system in plasma clot originally described by Ludwig et al. We tested the effects of the addition of phytohemagglutinin (PHA), coupled with a transformation of the original plasma clot method into a liquid culture system. A statistically higher number of myeloma colonies was observed in the liquid system in the presence of PHA (20 cases, median 84.5 vs. 9.5; p = 0.005), whereas a single variant (either PHA alone or liquid system alone) did not determine any significant growth variation. The increase in the cloning efficiency was evident even in the cases characterized by low bone marrow plasma cell infiltration, suggesting that this method is suitable for the described purposes. PMID- 3053935 TI - [Analysis of clinical results in infant cardiac surgical context of TGA and TAPVC in terms of cardioplegia]. PMID- 3053936 TI - [Monostotic fibrous dysplasia of rib bone]. PMID- 3053937 TI - [Acute purulent pericarditis followed by early constriction--report of three cases]. PMID- 3053938 TI - [A case of myasthenia gravis associated with rheumatoid arthritis, Sjogren's syndrome, hypergammaglobulinemia and hyperthyroidism]. PMID- 3053939 TI - [A case of solitary bronchial papilloma and a review of the literature]. PMID- 3053940 TI - Evaluation of the clinical pharmacological activity of a phlebotonic agent. Application to the study of Daflon 500 mg. AB - Provided certain methodological precautions are observed, mercury strain gauge venous occlusion plethysmography represents one of the best available function tests in phlebology to evaluate the efficacy of a phlebotonic agent. All physiological and environmental parameters liable to affect venous tone must be determined and controlled before and during the procedure, which is to be carried out in controlled and fully reproducible conditions. Intra and inter-observed reproducibility of results must be checked regularly in each laboratory. The objective character of the parameters assessed (Hmax, delta Vmax, T50, T2p...) does not spare the need for randomized, crossover or comparative, double-blind placebo-controlled trials, as plethysmographic results may show wide variations from one subject to another, and even from one moment to another, in chronic venous insufficiency, depending on the subject's environment and occupation. Furthermore, knowledge of treatment by the observer may affect the validity of the results. Taking into account all of these methodological pre-requisites, the phlebotonic activity of a single dose of Daflon 500 mg in lower limb chronic venous insufficiency was assessed in placebo-controlled clinical pharmacological studies. The following characteristics emerged: (1) significant improvement of venous capacitance, distensibility, and emptying times, as from the 2nd hour following Daflon 500 mg administration; (2) dose-response relationship, with an optimal dose of 2 tablets per day; (3) rapid onset and long duration of action; (4) significant hemodynamic effect on the venous system, whatever the type of venous insufficiency: organic, functional or gestational. PMID- 3053941 TI - Study of capillary filtration by double labelling I131-albumin and Tc99m red cells. Application to the pharmacodynamic activity of Daflon 500 mg. AB - The Tc99m albumin test can be used to confirm idiopathic cyclic oedema (ICO) syndrome by a pathological retention of albumin of more than 8% of the maximum level, following release of the tourniquet (results obtained in 420 patients compared with 100 healthy women). This test was completed by a study involving double labelling using I131-albumin and Tc99m red cells in ten healthy subjects and ten female patients with untreated ICO syndrome. The tests were repeated at a six week interval. Evaluation of retention levels after tourniquet release showed nil retention of red cells in both populations and nil albumin retention in the healthy subjects whilst it was pathological in the patients with ICO. The curve of decreasing radioactivity was often marked by irregular oscillations after tourniquet removal. Study of these oscillations by the Fast Fourier Transform revealed reproducible abnormalities in the low frequency zone only, between 37 and 28 mHz in the ICO patients. These abnormalities took the form of more frequent peaks with higher amplitudes than in the healthy subjects and were linked--in the case of excessive leak of albumin from capillaries--to lymphatic resorption. In the high frequency zone between 630 and 39 mHz, results corresponding to variations related to arteriovenous compliance were similar in both populations and by both tests. A double blind placebo-controlled trial in 30 patients with ICO was carried out to study the pharmacodynamic activity of a flavonoid, Daflon 500 mg (2 tabs daily for 6 weeks), which revealed a decrease in the degree of retention--initially high--of labelled albumin (p = 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3053943 TI - [The lethal and kinetic effects of carboplatin studied using the human tumor clonogenic assay and flow cytometry]. PMID- 3053942 TI - Clinical evaluation of a venotropic drug in man. Example of Daflon 500 mg. AB - Chronic venous insufficiency (CVI) of the lower limbs is a complex and fluctuating disease by its pathogenic mechanisms and its clinical symptoms. Although symptoms are subjective, they affect the quality of life and socio professional activity of many patients. This is why convincing demonstration of therapeutic activity of a venotropic drug should be carried out according to strict methodology. Only randomized double-blind controlled trials versus placebo (no reference drugs being available) could demonstrate the activity on condition that they are set up in a protocol, the statistical design of which, is adapted and defined "a priori". Inclusion, non inclusion and judgement criteria must be rigorous, taking into account many exogenous and endogenous factors which could have influence on the severity or the change in CVI and on the comparability of groups at the beginning and at the end of the study. Thus, the main risk factors of CVI (heredity, obesity, obstetrical and gynecological history, estroprogestogen treatment, profession, environment, etc.) and the season when the patient is recruited should be taken into account. With respect to all these restraints on methodology, the venotrotopic activity of a flavonoid Daflon 500 mg (2 tablets daily) was demonstrated in 200 patients (174 women, 26 men) with organic CVI (n = 83) or functional CVI (n = 117) treated for two months in two double-blind randomized trials versus placebo. The venotropic activity of Daflon 500 mg, was shown by a significant reduction of CVI signs and symptoms, whether organic or functional, and a significant improvement in venous hemodynamics according to plethysmographic parameters. Good acceptability was observed after medium and long term trials. PMID- 3053944 TI - [The beneficial effect of human urinary colony-stimulating factor (P-100) on recovery from granulocytopenia after anti-cancer chemotherapy for gynecologic cancer]. PMID- 3053946 TI - Microtia: reconstruction of the auricle. PMID- 3053945 TI - Female genital prolapse and pelvic floor deficiency. PMID- 3053947 TI - Stab wound of the gravid uterus: a case report and literature update. PMID- 3053949 TI - John Shaw Billings (1838-1913). PMID- 3053948 TI - Effect of insulin immune complexes on human blood monocyte and endothelial cell procoagulant activity. AB - Insulin antibodies and immune complexes (ICs) are present in most patients with diabetes after initiation of conventional insulin therapy. We studied the ability of bovine insulin, porcine insulin, and human insulin ICs to stimulate the procoagulant activity (PCA) of human blood monocytes (HBMs) and human umbilical vein endothelium (HUVE) in vitro. Polyclonal insulin antibodies from patients with insulin antibody-mediated resistance were isolated by immunoabsorption. PCA expressed by isolated adherent HBMs or cultured HUVE was quantified by radioimmunoassay of fibrinopeptide A with a 1:80 dilution of human plasma. Beef, pork, and human insulin, in quantities determined from titration curves, were added to fixed amounts of antibody to obtain equivalent amounts of ICs. For four different antibodies, the ratios of PCA (expressed by HBMs exposed to ICs or to antibodies alone and normalized for cell numbers) to activity expressed by tissue culture medium were as follows: beef insulin ICs, 2.55 +/- 0.41; por insulin ICs, 1.47 +/- 0.16; human insulin ICs, 1.28 +/- 0.19; and antibody, 1.28 +/- 0.32. The differences between beef and pork insulin ICs and antibody controls were significant. For a fifth antibody, a dose response between beef insulin ICs and PCA expression was demonstrated. For a sixth antibody, HBMs could express PCA when stimulated with antibody of a relatively low binding capacity (0.35 ng bovine insulin per microgram of antibody) only in the presence of lymphocytes. Conditioned medium from these cells significantly stimulated endothelial cell procoagulant activity. These observations raise the possibility that insulin ICs, that is, beef insulin ICs, may generate increased PCA and thereby contribute to the vascular complications of diabetes mellitus.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3053950 TI - Children and adolescents with traumatic brain injury: impact on the family. PMID- 3053951 TI - Acquired cerebral trauma: epilogue. PMID- 3053952 TI - Decrease in pyrogenicity of muramyl dipeptide after coupling with luteinizing hormone-releasing hormone. AB - Muramyl dipeptide (MDP) and its adjuvant active derivative lysine-MDP (Lys-MDP) have been demonstrated to be pyrogenic and to induce endogenous pyrogen (EP) production in vivo and in vitro. It has recently been shown that immunologic castration can be achieved in mice by immunization with luteinizing hormone releasing hormone (LHRH) directly conjugated by carbodiimide to Lys-MDP, termed LHRH-Lys-MDP (cdi), or with a linear monomeric MDP-linked molecule obtained by total synthesis, termed LHRH-Lys-MDP (s). These preparations were tested in the rabbit for their capacity to induce fever and were found to be devoid of pyrogenicity at dosage levels of Lys-MDP that induced fever. This decrease of pyrogenicity of Lys-MDP after coupling to LHRH seems to be related to the structure of the conjugate because the derivative LHRH-LysNH2-MDP exhibited the same pyrogenic activity as the free glycopeptide. Surprisingly, nonpyrogenic LHRH Lys-MDP induced production of EP and interleukin-1 (IL-1) in vitro and increased in vivo modifications of metal levels attributed to the action of IL-1. Moreover, LHRH-Lys-MDP reduced the pyrogenic effect of an exogenous dose of EP. PMID- 3053954 TI - Certified scuba diver volunteers at National Aquarium. PMID- 3053953 TI - Granulocyte-macrophage colony stimulating factor (GM-CSF) and macrophage colony stimulating factor (CSF-1) synergize to stimulate progenitor cells with high proliferative potential. AB - The ability to expand a population of bone marrow progenitors capable of forming macrophage colonies of high proliferative potential (HPP-CFC) was achieved using a culture system and signal requirements not previously shown to support the growth of HPP-CFC. Using bone marrow cells from untreated animals (not 5-FU treated), culture conditions designed primarily for the detection of GM-CFC or M CFC progenitors, and a more stringent criteria for HPP-CFC colony size (greater than or equal to 2 mm diameter), HPP-CFC progenitor expansion was demonstrated following simultaneous addition of rGM-CSF and CSF-1 to bone marrow cultures. Examination of the sequence and temporal requirements for rGM-CSF and CSF-1 addition necessary for the development of HPP-CFC-like colonies revealed that addition of the second factor could be delayed for up to 5 days and still result in the development of significant numbers of HPP-CFC colonies. Based on a comparison with human spleen cell conditioned medium (HSCM) and interleukin 1 (rIL 1), as sources of "synergistic activity" (SA) for the development of HPP-CFC like colonies in combination with CSF-1, the combination of GM-CSF and CSF-1 appears to represent a novel pathway for stimulating the expansion of HPP-CFC progenitors with high proliferative potential. PMID- 3053955 TI - Rationale for treatment of cancer with calcium antagonists. A review. AB - Calcium antagonists are broad spectrum antineoplastic agents capable of influencing favorably the course of cancer by inhibiting growth factor-initiated cell activation, blocking metastasis, or reversing inherent or acquired resistance to chemotherapeutic drugs. Calcium antagonists constitute an attractive category of agents for clinical trials in humans because their effects on malignancy in vivo in experimental animals are dramatic, and because there are several such agents on the market that can be adapted readily for such studies. However, an appreciation of the mechanisms of action of these drugs on cancer is essential for selection of an experimental design for therapeutic trials that will test these mechanisms adequately. The purpose of this review is to summarize available information on effects of calcium antagonists on cancer to facilitate incorporation of such drugs into therapeutic trials in human malignancy. PMID- 3053956 TI - The use of obstetric ultrasonography: the first 1,000 examinations at Ramathibodi Hospital. PMID- 3053957 TI - Hypercholesterolaemia and thyroid hormone status. PMID- 3053958 TI - The role of growth hormone in diabetes mellitus. AB - The insulin and growth hormone (GH)/insulin-like growth factor-I (IGF-I) axis are two endocrine systems that are interlinked at many levels. GH is one of the glucose counter-regulatory hormones, rising in response to hypoglycaemia, it has both intrinsic hyperglycaemic actions and causes insulin resistance. Both IGF-I and its receptor have high structural and functional homology to insulin and its receptor. Insulin can regulate IGF-I production, acting on the GH receptor or at a post-receptor site. Conversely IGF-I is thought to have a permissive effect on the pancreatic insulin response to glucose. Growth is compromised in poorly controlled diabetic children; however, a causal link with altered GH/IGF-I levels has not been proven. Insulin-dependent diabetes clearly causes derangements in the GH/IGF-I axis. In poorly controlled diabetics GH levels are invariably raised whilst normal or low levels of IGF-I are found, indicating a dissociation between the two factors. Altered IGF-binding protein levels are also found, with high levels of small binding protein and low levels of large binding protein. These derangements are probably the result of interactions at many levels although the exact mechanisms are not fully understood. Raised GH levels could result from altered hypothalamic/pituitary control or reduced feedback inhibition. The latter could, in turn, result from low IGF-I levels, reduced availability of IGF-I to relevant receptors or increased levels of inhibitors (possibly the small binding protein). Low IGF-I levels could be directly due to deficient insulin levels or simply to lack of available circulating binding protein. Alternative or altered molecular forms of circulating GH in diabetes seem unlikely on present evidence. That GH has an effect on glycaemic control is most evident from the abnormal glucose tolerance seen in acromegalics, but is also seen with physiological GH variations such as during the pubertal growth spurt. In diabetics the derangements to the GH/IGF-I axis, caused by poor metabolic control, leads to aggravation of the metabolic problems. Altered GH/IGF-I levels have been implicated in the long-term complications associated with diabetes, and whilst GH/IGF-I are not essential for the early changes involved in these complications they may still play an important role in their development, especially proliferative retinopathy. PMID- 3053959 TI - One-step purification of chicken growth hormone from a crude pituitary extract by use of a monoclonal immunoadsorbent. AB - The immunization of mice with an affinity-purified glycoprotein preparation from chicken pituitary tissue yielded several monoclonal antibodies towards the recently described glycosylated variant of chicken GH. As all these antibodies recognize the classical (non-glycosylated) GH molecule equally well, they provide a suitable tool for the development of both a specific immunoadsorbent and an assay method. This paper deals with the surprising purification power of the immunoadsorbent that was produced with one of the monoclonal antibodies. The resulting preparation was more than 99% pure as assessed by reversed phase high performance liquid chromatography and sodium dodecyl sulphate-polyacrylamide gel electrophoresis, so that no further purification steps were needed before the determination of the amino acid sequence of the material. The efficiency of the purification protocol as determined by a homologous, monoclonal antibody-based radioimmunoassay was virtually absolute. Moreover, the affinity-purified GH preparation was a mixture representing the multiple molecular forms of pituitary chicken GH, including both oligomeres and glycosylated GH. The purified preparations were finally used to demonstrate the hepatic 5'-monodeiodinase stimulating activity of GH in the chicken embryo (results not shown), in order to prove that the biological activity of the molecule had not been damaged by elution from the immunoadsorbent. PMID- 3053960 TI - The influence of plasma concentrations of tri-iodothyronine on the acute increases in insulin and muscle protein synthesis in the refed fasted rat. AB - To examine whether the low plasma levels of triiodothyronine (T3) in fasted rats might limit the recovery of muscle protein synthesis on refeeding, rats were fasted for either 3 or 4 days and refed with or without pretreatment with thyroid hormones. Fasting suppressed T3 levels, plasma insulin and the rate of the translational phase of muscle protein synthesis (KRNA; the rate per unit RNA), especially after the 4-day fast. On refeeding, plasma T3 levels remained low for more than 3 h after the 3-day fast and for more than 8 h after the 4-day fast. Insulin concentrations increased within the first hour of refeeding, eventually achieving supranormal concentrations after the 3-day fast. The KRNA increased within the first hour of refeeding, achieving well-fed control values by 3 h after the 3-day fast or 24 h after the 4-day fast. The increases in KRNA were significantly correlated with the increases in insulin at low insulin concentrations, achieving a plateau value at 150 pmol/l, so that further increases in insulin were not associated with any further increases in protein synthesis. Pretreatment with thyroid hormone induced increased T3 levels which were maintained for up to 8 h of refeeding. This had no effect on the responses of either insulin or protein synthesis to refeeding after the 3-day fast, but did result in an acceleration of the recovery in the KRNA and plasma insulin levels in the rats fasted for 4 days. Analysis of the insulin-KRNA relationship showed no evidence for any increase in the insulin sensitivity of muscle protein synthesis with thyroid pretreatment, the initial stimulation of protein synthesis on refeeding the rats fasted for 4 days reflecting increased insulin secretion. Since in the untreated animals, insulin secretion on refeeding was also correlated with T3 levels, these results are consistent with the previously reported thyroidal dependence of insulin secretion. PMID- 3053961 TI - Solid-phase iodothyronine-5'-deiodinase (5'-D) assays applied in production of monoclonal antibodies against 5'-D. AB - Characterization of iodothyronine-deiodinating enzymes has been difficult due to loss of enzyme activity during purification. To obtain a new tool for studying these enzymes we investigated the possibility of developing monoclonal antibodies (MAbs) against iodothyronine-5'-deiodinase (5'-D). Two specific and sensitive solid-phase microassays were developed for screening hybridoma supernatants for the presence of antibodies inhibiting rat kidney 5'-D and antibodies binding to but not inhibiting the enzyme. BALB/c mice were immunized with a 3-((3 cholamidopropyl)-dimethylammonio)-1-propanesulphonate (CHAPS)-solubilized 5'-D rich membrane preparation from rat kidney cortical tissue. Spleen cells were fused with NSI-Ag 4/l mouse myeloma cells by means of polyethylene glycol. Two hybridoma cell lines (AF5 and BE8) secreting MAbs specifically binding to without inhibiting 5'-D were produced. The AF5 antibody was of the IgG2a subclass and the BE8 antibody of the IgG2b subclass. Binding of one of the antibodies to the enzyme inhibited binding of the other in both an enzyme-linked immunosorbent assay (ELISA) and a specific enzyme-binding assay. CHAPS-solubilized kidney microsomal fraction was chromatographed on a Sepharose 6B column. Elution profiles of 5'-D activity and MAb-binding antigens, as measured by ELISA with both AF5 and BE8, were identical. Monoclonal antibodies should be valuable probes in the further elucidation of the nature of the iodothyronine-deiodinating activity in various tissues. PMID- 3053964 TI - Fertilization of zona-drilled mouse oocytes treated with a monoclonal antibody to the zona glycoprotein, ZP3. AB - Opening a small aperture in the zona pellucida of mouse oocytes by using micromanipulation and a stream of acidified Tyrode's solution (zona drilling) improved the efficiency of in vitro fertilization at low sperm concentrations without adversely affecting development to the blastocyst stage. Zona drilling also permitted in vitro fertilization and development when sperm penetration through the zona was blocked by a monoclonal antibody to the protein core of the zona glycoprotein, ZP3. These results provide a direct demonstration that sperm entry occurs through the aperture and also suggest that zona drilling of human oocytes may offer a therapeutic approach when autoantibodies to the zona pellucida are suspected as a cause of infertility. PMID- 3053963 TI - Isolation and sequence analysis of serine protease cDNAs from mouse cytolytic T lymphocytes. AB - Three new cDNA clones (designated MCSP-1, MCSP-2, and MCSP-3) encoding mouse serine proteases were isolated from cloned cytolytic T lymphocytes (CTL) by a modified differential screening procedure. The putative mature proteins of MCSP-2 and MCSP-3 are each composed of 228 amino acids with molecular weights of 25,477 and 25,360, respectively. NH2-terminal amino acids of MCSP-2- and MCSP-3 predicted proteins were identical to those reported for granzyme E and F, respectively. The third species, MCSP-1, was closely related to the two other cDNA species but approximately 30 amino acids equivalents of the NH2-terminal portion of the cDNA were not cloned. The amino acids forming the active sites of serine proteases were well conserved among the three predicted proteins. The active site pocket residue positioned six residues before the active-site Ser184 is alanine in MCSP-1, threonine in MCSP-2, and serine in MCSP-3, indicating that both MCSP-2 and MCSP-3 may have chymotrypsin-like specificity. There are three potential asparagine-linked glycosylation sites in MCSP-1 and MCSP-3, and four in MCSP-2-deduced amino acid sequences. Amino acid comparison of MCSP-1 with four other reported serine proteases whose active site pocket residue is alanine revealed that MCSP-1 was substantially different from the other molecules, indicating that MCSP-1 may be a new member of mouse T cell serine protease family. Antibodies made against a MCSP-1 lacZ gene fusion protein stain granules of CTL and react on immunoblots with two distinct granule protein bands of 29 and 35-40 kD. Only the 35-kD species labels with [3H]DFP. Since a protease cascade may play a key role in cytolytic lymphocyte activation, our isolation of cDNAs representative of unique serine esterases should help to investigate such a cascade process. PMID- 3053962 TI - The nature and kinetics of a delayed immune response to purified protein derivative of tuberculin in the skin of lepromatous leprosy patients. AB - We have analyzed the nature and kinetics of a delayed, cell-mediated immune response to a purified protein derivative of tuberculin (PPD) in the skin of 154 naturally sensitized patients with lepromatous leprosy. After the intradermal injection of 5 U of PPD, biopsies were taken at 1-21 d and studied for the composition, extent, persistence, and organization of the emigratory cell response by light and electron microscopy. Induration of positive sites occurred promptly, reached a maximum diameter at 4 d, displayed a major extravasatory element, and was evident for as long as 21 d. The cellularity of the site exhibited a biphasic course, reached a maximum at 7 d, involved as much as 70% of the dermis and millions of new cells, and was elevated threefold above preinjection levels at 21 d. The emigratory cells were limited to T cells and circulating monocytes. T cells were more evident as they entered a preexisting lepromatous lesion containing parasitized macrophages and only occasional T cells many of the CD8+ phenotype. The predominant emigratory T cell was CD4+ although CD8+ cells were in evidence. The CD4/CD8 ratio of the lesions started at less than unity and in two distinct steps reached levels as high as 5:1. In most sites CD4+ cells were in the majority at 21 d. A well-defined granulomatous response with epithelioid and giant cells was apparent at 4 d, reached a maximum at 7 d, and involved all PPD sites at this time point. The generation of these differentiated mononuclear phagocytes from newly emigrated monocytes was never observed in the underlying lepromatous lesion but is a constant feature of the tuberculoid leprosy response. Epidermal thickening and keratinocyte proliferation, sequellae of the dermal reaction, reached a maximum at 7 d and gradually resolved by 3 wk. A constant feature of the PPD response was the extensive destruction of preexisting macrophages containing Mycobacterium leprae bacilli or their products. This was associated with the presence of and intimate contact with highly polarized lymphoid cells of unknown phenotype. Cell destruction did not involve other elements of the dermis and spared parasitized Schwann cells. Newly emigrated T cells and monocytes were never seen within the perineural sheath in contact with neural elements. It appears that a single antigenic stimulus leads to a very long-term, defined series of events with distinct temporal patterns. It includes waves of emigratory T cells, the maturation and organization of monocytes, the generation of killer cells, and the extensive destruction of parasitized macrophages.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 3053965 TI - Changes in pituitary responsiveness during the ovulatory cycle of the Japanese quail, in vitro. AB - Anterior pituitary glands from ovulating Japanese quail (Coturnix coturnix) were used to investigate variation in sensitivity to chicken luteinizing hormone releasing hormone (cLHRH I; Gln8-LHRH). Grouping the pituitaries by ovulatory stage provided preliminary evidence of changes in sensitivity to LHRH during the ovulatory cycle. Pituitaries taken from quail before the preovulatory LH surge were responsive to cLHRH I, while pituitaries from the other times of the cycle showed minimal response to cLHRH I. Female pituitary glands release less LH than those of males. These data indicate a change in sensitivity to LHRH in the female quail that may be due to changes in gonadal steroids or the pool of releaseable LH from the pituitary. PMID- 3053966 TI - Development of beta-amino acid transport in the kidney. AB - This study focuses on the maturation of the renal beta-amino acid transport system and uses dietary manipulation as a probe. The epithelial surface of the renal proximal tubule is responsible for the conservation of ions and organic solutes including beta-amino acids. This beta-amino acid transport system is stimulated during periods of reduced dietary intake and permits increased excretion following dietary excess. We have examined transport of the sulfur containing beta-amino acid, taurine, as a measure of this renal adaptive response to fluctuations in dietary sulfur amino acid intake and as a substrate for the beta-amino acid transport system. A precession of taurine uptake values by brush border membrane vesicles (BBMV) prepared from nursing rats from youngest to oldest was evident. However, these membranes demonstrate the full renal adaptive response to altered sulfur amino acid intake after the first week of life. This adaptive response is expressed at the brush border surface by transport changes in both directions ("up regulation" and "down regulation"), through changes in the initial rate (15 sec) of Na+-taurine cotransport. No alterations in the lipid microenvironment of the membrane, as detected by altered membrane fluidity, were uncovered. Although vesicles from 7-day-old pups demonstrate adaptation and accumulate taurine to a limited extent, the accumulation of Na+, which energizes uptake, may be altered, thereby preventing full expression of the adaptive response and of transport capacity at this age. PMID- 3053968 TI - Suicide prevention update: thirty years later. PMID- 3053967 TI - PROs: an update. PMID- 3053970 TI - Studies on the proteolytic activity of Bacteroides fragilis. AB - The proteolytic activity of the intestinal bacterium Bacteroides fragilis NCDO 2217 was cell-bound during exponential growth, but was progressively released from the cells in stationary phase. Proteins hydrolysed included casein, trypsin, chymotrypsin, azocasein and the proteins in azosoya bean flour. Collagen, azocoll, elastin, gelatin, ovalbumin and bovine serum albumin were either weakly degraded or completely refractory to proteolysis. Arylamidase activity was exhibited against leucine p-nitroanilide (LPNA), leucine beta-naphthylamide, glycyl-proline p-nitroanilide and valyl-alanine p-nitroanilide. The bacterium grew with ammonia, peptone or casein as sole nitrogen source. Azocasein- and LPNA hydrolysing activities were consistently higher when grown on casein. Cell-bound protease activity increased concomitantly with growth rate in both carbon- and nitrogen-limited continuous culture. Leucine arylamidase activity was also growth rate-dependent, being 3-fold greater at D = 0.18 h-1 compared to D = 0.03 h-1. Extracellular proteolytic activity was only detected at low growth rates, accounting for about 25% of total protease activity. PMID- 3053969 TI - In vivo production of a tissue-destructive protease by Legionella pneumophila in the lungs of experimentally infected guinea-pigs. AB - A tissue-destructive protease of Legionella pneumophila was assayed for in the lungs of experimentally infected guinea-pigs by ELISA. It was found in amounts equivalent to the known lethal dose of purified protease administered by the intranasal route. The identity of the protease was confirmed by immunoblot analysis. This is further evidence that Legionella pneumophila protease may play a major role in the pathogenesis of Legionnaires' disease. PMID- 3053971 TI - Carrot phytoalexin alters the membrane permeability of Candida albicans and multilamellar liposomes. AB - The biochemical basis for the antimicrobial effect of the carrot phytoalexin 6 methoxymellein (6-MM) was examined. At fungistatic concentrations 6-MM retarded the ability of Candida albicans to incorporate radioactive thymidine, uridine and leucine into biopolymers. When C. albicans was incubated with 6-MM, 260-nm absorbing materials and 3H-labelled compounds leaked from the cells. The inhibitory effects of 6-MM on cell growth and membrane functions were, however, reduced as the concentration of divalent metal cations added to the medium was increased. 6-MM interacted with multilamellar liposomes constituted from phosphatidylcholine, cholesterol and dicetyl phosphate, or from phosphatidylcholine only, resulting in the release of glucose trapped in these liposomes. These results suggest that 6-MM exerts its toxic effects on susceptible cells as a result of its interaction with their membranes and disturbance of membrane-associated functions. PMID- 3053972 TI - Motility of Campylobacter jejuni in a viscous environment: comparison with conventional rod-shaped bacteria. AB - The motility of four strains of Campylobacter jejuni in solutions of varying viscosity was measured and compared to that of a number of conventional rod shaped bacteria (CRSB). All the bacteria tested showed an initial increase in velocity in the low viscosity solutions--between 1 and 3 centipoise (1 P = 0.1 Pa s). However, only the campylobacters were actively motile in highly viscous solutions with velocities ranging from 60 to 100 micron s-1. All strains of C. jejuni tested showed three separate peaks of motility as the viscosity of the solution was increased. A higher proportion of C. jejuni cells exhibited longer path lengths when the viscosity of the surrounding medium was increased from 1.4 to 57 cP. The findings of the study suggest that C. jejuni has a motility suited to movement in a viscous environment, and that this ability might provide the organism with an ecological advantage when in intestinal mucus. It is proposed that the mechanism of motility changes depending on the viscosity of the supporting environment. PMID- 3053973 TI - Cloning, expression in Escherichia coli and nucleotide sequence of a tetracycline resistance gene from Streptomyces rimosus. AB - Determinants of tetracycline resistance have been cloned from two different tetracycline-producing industrial strains of Streptomyces into Streptomyces lividans using the plasmid vector pUT206. Three plasmids, pUT250 and pUT260 with a 9.5 and a 7.5 kb insert respectively of Streptomyces rimosus DNA, and pUT270 with a 14.0 kb insert of Streptomyces aureofaciens DNA, conferring resistance to tetracycline, have been isolated. By in vitro sub-cloning, a similar fragment of 2.45 kb containing the tetracycline resistance gene (tet347) was further localized on these plasmids. The S. rimosus gene has been cloned into Escherichia coli and expressed under the control of lambda pL or Lpp promoters. Differential protein extraction of E. coli cells revealed the presence of an additional membrane-embedded protein in tetracycline-resistant cells. On the basis of available restriction endonuclease maps, the tet347 gene is probably identical to the tetB gene from S. rimosus recently identified by T. Ohnuki and co-workers as responsible for the reduced accumulation of tetracycline. The nucleotide sequence of a 2052 bp DNA fragment containing the TcR structural gene from S. rimosus has been determined. The amino acid sequence of the tet347 protein (Mr35818) deduced from the nucleotide sequence shows a limited but significant homology to other characterized tetracycline transport acting determinants from pathogenic bacteria. PMID- 3053974 TI - Accumulation of LamB-LacZ hybrid proteins in intracytoplasmic membrane-like structures in Escherichia coli K12. AB - The subcellular location of LamB-LacZ hybrid proteins in the Escherichia coli K12 strains pop3234 and pop3299 was investigated by immunocytochemical detection and protease-accessibility experiments. Induction of the synthesis of the hybrid proteins resulted in the appearance of membrane-like structures within the cytoplasm of the cells. Labelling of ultrathin cryosections of the cells with anti-beta-galactosidase or anti-LamB protein serum and protein-A-gold complexes revealed that the hybrid proteins were associated with these membrane-like structures or accumulated within the cytoplasm. Protease-accessibility experiments confirmed this localization. Moreover, when low quantities of hybrid proteins were produced, i.e. in uninduced pop3234 cells or in induced pop3299 cells, the hybrid proteins were accessible to trypsin from the periplasmic side of the inner membrane, leaving protected fragments with an apparent Mr of 83,000. Apparently, these hybrid proteins are partly translocated through the inner membrane, resulting in membrane-spanning forms of the proteins. PMID- 3053975 TI - Glycerol uptake mutants of the hyphal fungus Aspergillus nidulans. AB - A new class of glycerol non-utilizing mutants, designated glcC, has been isolated. The glcC gene was mapped in linkage group VI and mutants were found to complement the reference strains glcA1 (linkage group V) and glcB33 (linkage group I) in diploids. The new mutants were unable to grow on glycerol. However, in contrast to the glcA and glcB phenotype these mutants did grow well on dihydroxyacetone and D-galacturonate. By in vivo 13C NMR spectroscopy it was shown that the glcC mutant did not take up glycerol but did take up dihydroxyacetone. The latter substrate was converted intracellularly into glycerol which was then catabolized as normal. PMID- 3053976 TI - Construction of cys:lac gene fusions in Escherichia coli and their use in the isolation of constitutive cysBc mutants. AB - Operon fusions of the lacZ gene to two different genes of the cysteine regulon controlled by the cysB regulatory protein were isolated. The fusion strains were used for selection of cysB constitutive mutants. Three cysBc alleles have been characterized and cloned into multicopy plasmids. PMID- 3053977 TI - Continued growth of Escherichia coli after stopping medium addition to a K+ limited chemostat culture. AB - The steady-state bacterial dry wt of Escherichia coli, growing under K+-limited conditions in the chemostat, was inversely dependent on the growth rate. This phenomenon was more carefully investigated in medium-flow stop experiments. Growth did not stop immediately but continued for a time, initially at the same rate as before. The dry wt increased to a value corresponding to a steady-state growth rate near zero, independent of the initial specific growth rate. This was observed in both the wild-type strain and a mutant that lacked the high-affinity K+ uptake system. The wild-type strain maintained a low extracellular K+ concentration both in the chemostat under steady-state conditions and after stopping the medium flow. The mutant, on the other hand, maintained a much higher extracellular K+ concentration in the steady state, which decreased to much lower values after stopping the medium flow. From the increase in bacterial dry wt and the low external K+ concentration after stopping the medium flow it is concluded that the intracellular K+ is redistributed among the cells, including new cells. The growth yield on K+ was highest in the stationary growth phase of a batch culture and all steady-state cultures converged ultimately to this yield value after the medium flow had been stopped. It is proposed that the growth rate of E. coli under K+-limited conditions is determined by the intracellular K+ concentration. PMID- 3053978 TI - Regulation of trehalase activity during the cell cycle of Saccharomyces cerevisiae. AB - Synchronous cultures of Saccharomyces cerevisiae prepared by selection of small unbudded cells from an elutriating rotor were used to measure trehalase activity during the cell cycle. After the small cells had been removed from the rotor, the remainder was used to prepare asynchronous control cultures. Both synchronous and control cultures were studied for two cell cycles. In asynchronous cultures the trehalase activity of crude cell lysates rose continuously. In synchronized populations trehalase activity increased from the beginning of budding onwards. However, around the period of cell division the enzyme activity dropped rapidly but transiently by more than 5-fold. The same changes were found during the second budding cycle. Measurements of invertase and glucose-6-phosphate dehydrogenase activities in the same synchronous and asynchronous cultures revealed a continuous increase for both enzymes. Incubation of cell lysates with cAMP-dependent protein kinase before assaying for trehalase resulted in a 2-fold enhancement of enzyme activity in asynchronous control cultures. In synchronized cells this treatment also led to a significant stimulation of trehalase activity, and largely abolished the cell-cycle-dependent oscillatory pattern of enzyme activity. These results suggest that the activity of trehalase during the cell cycle is regulated, presumably at the post-translational level, by a phosphorylation-dephosphorylation mechanism. PMID- 3053979 TI - Vero cytotoxin production and presence of VT genes in Escherichia coli strains of animal origin. AB - Vero cytotoxin (VT) production has been studied in 34 Escherichia coli strains isolated from animals with enteric diseases. The strains were tested by DNA hybridization with probes for VT1 and VT2 sequences and also in toxin neutralization experiments with specific antisera. Twenty bovine strains, belonging to nine different O serogroups, produced VT1 or VT2 but not both toxins. In contrast, all 14 porcine strains of four different O serogroups produced VT2 only. Six of these porcine strains, belonging to serogroups O138, O139 and O141, were isolated from cases of oedema disease. In general, the porcine isolates produced toxin at a lower level than the bovine strains. PMID- 3053980 TI - Role of lipopolysaccharide and complement in susceptibility of Escherichia coli and Salmonella typhimurium to non-immune serum. AB - The role of lipopolysaccharide (LPS) in the susceptibility of Escherichia coli and Salmonella typhimurium to non-immune human serum was investigated using serum sensitive strains of both enterobacteria. LPS from serum-resistant strains of E. coli and S. typhimurium could activate and completely remove the serum bactericidal activity, and also showed dose-dependent anti-complement activity. These properties were mainly due to the high-molecular-mass LPS: the low molecular-mass LPS from serum-resistant strains of E. coli and S. typhimurium had only a slight effect on the serum bactericidal activity, and showed only low anti complement activity, even at high concentration. The results suggest that LPS composition, especially the O-antigen polysaccharide chains, contributes to the susceptibility of E. coli and S. typhimurium strains to complement-mediated serum bactericidal activity. PMID- 3053981 TI - Low concentrations of trifluoperazine arrest the cell division cycle of Saccharomyces cerevisiae at two specific stages. AB - Low concentrations of trifluoperazine (TFP) reversibly inhibited vegetative growth of Saccharomyces cerevisiae. The cell cycle was analysed by flow cytometry using haploid a cells synchronized by alpha-factor arrest and several temperature sensitive cell division cycle mutants (cdc). Cells were pulse-labelled with fluorescein-labelled concanavalin A (ConA-FITC) to determine cell division or stained with propidium iodide to determine the stage of cell cycle arrest by TFP. Cell growth was estimated from the changes in the relative intensity of scattered light, and budding was determined microscopically. When TFP was added before Start on release from alpha-factor arrest, after release of cdc28-arrested cells, and at transition from stationary phase to vegetative growth, cell growth, budding and DNA synthesis were inhibited. When TFP was added after execution of spindle pole body duplication, cell growth, bud emergence and DNA synthesis were not inhibited but cell division was inhibited and the cells arrested with buds at G2M Using cdc mutants, the second stage of arrest by TFP was determined to be just before medial-nuclear division. PMID- 3053982 TI - Isolation and characterization of mevinolin resistant mutants of Saccharomyces cerevisiae. AB - Mutant of Saccharomyces cerevisiae resistant to mevinolin, a competitive inhibitor of 3-hydroxy-3-methylglutaryl-coenzyme-A (HMGCoA) reductase (EC1.1.1.34) were isolated and one mutant (MV71) was extensively characterized. While growth of resistant strains in the presence of mevinolin was growth. Diploids produced by mutant/wild-type matings showed levels of mevinolin resistance which indicated incomplete dominance. Sterol synthesis in the presence of mevinolin was inhibited in strain MV71 but to a lesser degree than seen in the wild-type strain. All mevinolin resistant mutants also demonstrated a slight resistance to the antibiotic nystatin. The subcellular location of HMGCoA reductase activity in MV71 and the wild-type strain were determined and it was shown that yeast HMGCoA reductase is not regulated by a dephosphorylation mechanism as has been shown for mammalian reductases. In vivo and in vitro studies of strain MV71 and the wild-type indicated that mevinolin resistance did not result in changes in HNGCoA reductase activity as has been demonstrated in mammalian systems. Based on growth data, sterol analysis, and the lack of detection of HMGCoA reductase activity differences between strain MV71 and the wild-type, mevinolin resistance is concluded to result possibly from a mutation in HMG2, one of the two functional yeast HMGCoA reductase genes, which accounts for a minor (up to 17%) amount of total cellular reductase activity. PMID- 3053983 TI - Further analysis of nitrogen fixation (nif) genes in Azotobacter chroococcum: identification and expression in Klebsiella pneumoniae of nifS, nifV, nifM, and nifB genes and localization of nifE/N-, nifU-, nifA- and fixABC-like genes. AB - The results presented extend previous investigations on the genetics of nitrogen fixation in Azotobacter chroococcum and indicate that nif- and fix-like DNA is located in at least five different regions of the genome. Region I contains functional copies of nifS,V and M, as well as nifH, D and K, all of which complemented mutants of Klebsiella pneumoniae. In addition, nifE- and/or nifN like and nifU-like DNA is located in this region. The organization of the nif cluster in region I closely resembles that of K. pneumoniae. though spread over 22 kb as compared with 14 kb. Region II contains a functional nifB gene, which complemented a K. pneumoniae nifB mutant, and seems to be adjacent to ap nifA like gene. Region III harbours nifH*, encoding a second nitrogenase Fe-protein. Region IV contains a reiteration of nifE- on and/or nifN-like sequences, and DNA homologous to Rhizobium meliloti fixABC is present in region V. The apparent complexity of nifDNA in A. chroococcum is probably related to the two systems for N2-fixation pr present in this organism. PMID- 3053984 TI - Comparison of mandelate dehydrogenases from various strains of Acinetobacter calcoaceticus: similarity of natural and 'evolved' forms. AB - In previous work it had been shown that Acinetobacter calcoaceticus wild-type strain NCIB 8250 had only an L-mandelate deydrogenase but it could give rise to mutants that contained an evolved D-mandelate dehydrogenase; conversely, wild type strain EBF 65/65 had only a D-mandelate dehydrogenase but gave rise to mutants that possessed an evolved L-mandelate dehydrogenase. Several other wild type strains of A. calcoaceticus have now been shown to grow on both enantiomers of mandelate. In every case the L-mandelate dehydrogenases were found to be much more heat-stable and insensitive to inhibition by p-chloromercuribenzoate than were the D-mandelate dehydrogenases when measured in bacterial extracts. All the D-mandelate dehydrogenases in the wild-type strains were inactivated to about the same extent by an antiserum that had been raised in a rabbit against an evolved D mandelate dehydrogenase. An evolved D-mandelate deydrogenase (from a mutant strain derived from strain NCIB 8250) and an original D-mandelate dehydrogenase (from a mutant strain derived from strain EBF 65/65) were purified to homogeneity by the same procedure and were indistinguishable as judged by immunological cross reactivity of the native and the sodium-dodecyl-sulphate-denatured enzymes, solubility in cholate, net charge at pH 7.5, pI value, salting-out properties, Mr value, apparent K(m) value for D-mandelate, heat-stability and sensitivity to p chloromercuribenzoate. The most likely explanation for the appearance of evolved mandelate dehydrogenases in strains of A. calcoaceticus is that cryptic genes become expressed. PMID- 3053985 TI - A common virulence region on plasmids from eleven serotypes of Salmonella. AB - Cured derivatives of Salmonella dublin and S. typhimurium showed reduced virulence following oral infection of mice (10(4)-10(5)-fold for S. dublin, 10(2) fold for S. typhimurium). Large plasmids from S. dublin and S. typhimurium independently restored virulence to the cured S. dublin but truncated S. dublin plasmids with deletions in a previously identified virulence region did not. This common virulence region identified in plasmids from S. dublin and S. typhimurium was shown to be carried on plasmids from 11 other serotypes of Salmonella but was absent from 10 plasmid-containing serotypes. TnA and Tn10 were transduced from the virulence region of two TnA-insertion mutants of S. dublin and one Tn10 insertion mutant of S. typhimurium that showed diminished virulence to recipient wild-type strains of S. dublin, S. enteritidis and S. typhimurium. Each transductant showed a decrease in mouse virulence within the range 10(3)-10(5). It is therefore proposed that similar virulence determinants are expressed in different serotypes. It was also shown that integration that occurred during curing was Tn10 dependent. PMID- 3053986 TI - A virus with a bisegmented double-stranded RNA genome in rat (Oryzomys nigripes) intestines. AB - Examination of the intestinal contents of free-living Oryzomys nigripes rats by PAGE revealed two sharply defined bands that could be stained by ethidium bromide or by silver nitrate with comparable intensities. The molecules forming these bands were susceptible to digestion by pancreatic RNase A but not by RNase T1 or by DNase I. Their lengths were estimated to be about 2.6 and 1.5 kbp, respectively, by comparison with rotavirus SA11 genome segments. They cosedimented in CsCl gradients at a density of 1.39 to 1.40 g/ml, together with uniform particles approximately 35 nm in diameter with indistinct surface structure. It is suggested that these particles represent an as yet undescribed virus with a bisegmented double-stranded RNA genome, for which the name 'picobirnavirus' is proposed. PMID- 3053987 TI - Co-expression of the hepatitis B surface and core antigens using baculovirus multiple expression vectors. AB - The hepatitis B (HB) virus DNA sequences coding for the pre-core (preC) or C antigens (HBpcAg, HBcAg) have been inserted into the baculovirus plasmid transfer vector, pAcYM1, such that the HB viral sequences are under the control of the polyhedrin promoter of Autographa californica nuclear polyhedrosis virus (AcNPV). Spodoptera frugiperda cells infected with either of the derived recombinant plasmids in the presence of infectious AcNPV DNA yielded recombinant, polyhedrin negative viruses that expressed high levels of the respective HBpcAg or HBcAg (representing approx. 5 to 10% and approx. 40% of the stained cellular proteins, respectively). The particulate 27 nm HBcAgs have been purified to homogeneity from infected cell extracts by density gradient centrifugation. Dual expression transfer vectors containing the HBcAg gene sequences and the coding sequences of the HB viral S antigen (HBsAg), each gene under the control of its own copy of the polyhedrin promoter, have also been constructed and used to derive recombinant viruses. The recombinant with the HB C and S genes expressed high levels of the HBcAg (approx. 40% of the cellular proteins) and low levels of the HBsAg (approx. 2% of the stained cellular proteins). Dual expression, occluded, recombinant baculoviruses that make HBsAg, as well as the AcNPV polyhedrin protein, have been prepared that are highly infectious for Trichoplusia ni caterpillars, allowing reproducible preparation of the antigen in larvae. Using radioimmunoassays (RIAs) and ELISAs, the recombinant HBcAg (RIA) and HBsAg (ELISA) have been used to identify human antibodies to HB virus with results that compare favourably with the data obtained with non-recombinant antigens. PMID- 3053988 TI - Characterization of Triatoma virus, a picorna-like virus isolated from the triatomine bug Triatoma infestans. AB - Some properties of Triatoma virus (TrV), a picorna-like virus recently isolated from Triatoma infestans, have been studied. Electron microscopic observations of purified viral preparations showed the presence of non-enveloped viral particles 30 nm in diameter. The sedimentation coefficient of virus particles was about 165S and the buoyant density in CsCl was 1.39 g/ml. The viral genome was composed of one single-stranded RNA molecule with an Mr of 3 x 10(6). Three major polypeptides with Mr values of 39K, 37K and 33K and a minor one of about 45K were found in the virus particle. TrV particles contain about 35% RNA and 65% protein by weight. These data support the classification of this virus in the family Picornaviridae. PMID- 3053989 TI - Efficient isolation of HIV from plasma during different stages of HIV infection. AB - Attempts to isolate human immunodeficiency virus (HIV) from blood plasma using inoculation of pellets from ultracentrifuged samples into cultures of peripheral blood mononuclear cells (PBMC) resulted in a high overall recovery rate (75%) of the virus from 76 patients in various stages of HIV infection. The recovery rate was dependent on the stage of infection; in patients with acquired immunodeficiency syndrome (AIDS) it was 100%, in AIDS-related complex (ARC) 86%, in persistent generalized lymphadenopathy (PGL) 64%, and in asymptomatic patients 54%. The HIV isolation rates compared favorably with those obtained after cocultivation of patient and target PBMC (overall recovery rate 67%). HIV was isolated from plasma but not from PBMC in 8 cases, whereas the reverse was true in 3 of 71 simultaneously tested cases. Isolation from plasma was found to be superior to detection of serum p24 antigen for the demonstration of HIV (positivity rates 75% and 30%, respectively). The time to appearance of p24 antigen in cultures inoculated with HIV-containing plasma samples was inversely related to the presence of detectable p24 antigen in serum. There was a significantly shorter time to culture positivity of plasma samples from AIDS and ARC patients than from PGL and asymptomatic patients. These results suggested that there is a progressive increase in the concentrations of infectious HIV in plasma from the asymptomatic to the AIDS stage. HIV isolation from plasma samples is a reliable means of demonstrating HIV viremia and has obvious advantages over the more commonly used cocultivation procedures. The frequent occurrence of cell free, infectious HIV in plasma suggests that the majority of HIV-infected patients have a relative lack of functional neutralizing antibodies against the virus, at least during the late stages of disease. PMID- 3053990 TI - Accentuated antibody response to paramyxoviruses in individuals infected with human immunodeficiency virus. AB - Sera from 31 human immunodeficiency virus (HIV)-infected patients, representing different clinical stages of HIV infection, were assayed for antibodies against measles and mumps viruses by various serological tests and compared to 23 healthy controls. Sera from four patients (two primary, one asymptomatic, and one acquired immunodeficiency syndrome) exhibited a pronounced antibody response to measles as detected by haemagglutination inhibition and radioimmuno-precipitation assay. The RIPA-positive sera showed increased reactivity to all the viral components and in particular to the haemagglutinin (HA) protein of the virus (Fig. 1). Three of these positive patients also showed a similar response to mumps virus. One of the control sera also showed an increase in antibody titre in measles serological tests. The measles antibodies were shown not be anti-HIV antibodies crossreacting with paramyxoviruses. The reactivity to haemagglutinin was still present when using nonglycosylated measles virus antigen grown in the presence of tunicamycin. Whether the accentuated antibody response is due to polyclonal activation mediated by HIV or to reactivation of the viruses remains to be answered. PMID- 3053991 TI - The renascence of neurotic depression: its varied dynamics and implications for outcome research. AB - Kraepelin's observation of an emotion-connected subgroup of depressions led to the division of the common clinical depressions into endogenous and neurotic. But although the connotation of the former was clear, the latter had no positive diagnostic features, with the result that it fell very largely out of use. This paper presents evidence that there are depressions that are a consequence of neurotic anxiety. They can be positively diagnosed, and are resolved by overcoming the underlying neurotic anxieties. The neurotic basis of many cases of depression explains the success of different psychotherapeutic strategies in alleviating nonpsychotic depression, since 50% of neurotic cases respond to the nonspecific impact of the therapeutic interview. Unfortunately, the various outcome studies are muddied by failure to realize that a variety of psychopathologies are represented in nonpsychotic depression. PMID- 3053992 TI - From Lavoisier to Freud. A historical epistemological note with contemporary significance. PMID- 3053993 TI - Chronic intracerebroventricular infusion of insulin failed to alter brain insulin binding sites, food intake, and body weight. AB - The present study was performed to explore the role of exogenous insulin in CSF in the control of energy balance in the rat. For this purpose, adult male Sprague Dawley rats carrying an indwelling cannula in the right lateral cerebral ventricle were infused for a maximum of 10 days with insulin (Actrapid) at various rates (starting at 0, 45, 85, 170, and 600 ng/day) or anti-insulin antibody (IgG fraction; diluted 1:10 wt/vol) with an osmotic minipump. All those treatments did not modify the growing rates; neither total daily food intake nor the circadian rhythm of food intake was further modified. The chronic insulin infusion starting at 600 ng/day resulted in a chronic significant increase in CSF insulin levels without changing the plasma insulin level. It failed to alter specific insulin binding sites to Triton X-100 solubilized microsomal membranes from various brain areas (cerebral cortex, olfactory bulbs, and lateral and medial hypothalami) at the end of the 5- or 10-day period of insulin infusion. Purification of insulin receptors on a wheat germ agglutinin did not reveal any further effect of insulin. From these results, it seems unlikely that the input to the brain insulin-effector systems could arise from CSF insulin. PMID- 3053994 TI - Neutral amino acid transport in astrocytes: characterization of Na+-dependent and Na+-independent components of alpha-aminoisobutyric acid uptake. AB - Neutral amino acid transport is largely unexplored in astrocytes, although a role for these cells in blood-brain barrier function is suggested by their close apposition to cerebrovascular endothelium. This study examined the uptake into mouse astrocyte cultures of alpha-aminoisobutyric acid (AIB), a synthetic model substrate for Na+-dependent system A transport. Na+-dependent uptake of AIB was characteristic of system A in its pH sensitivity, kinetic properties, regulatory control, and pattern of analog inhibition. The rate of system A transport declined markedly with increasing age of the astrocyte cultures. There was an unexpectedly active Na+-independent component of AIB uptake that declined less markedly than system A transport as culture age increased. Although the saturability of the Na+-independent component and its pattern of analog inhibition were consistent with system L transport, the following properties deviated: (1) virtually complete inhibition of Na+-independent AIB uptake by characteristic L system substrates, suggesting unusually high affinity of the transporter; (2) apparent absence of trans-stimulation of AIB influx; (3) unusually concentrative uptake at steady state (the estimated distribution ratio for 0.2 mM AIB was 55); and (4) susceptibility to inhibition by N-ethylmaleimide. Direct study of the uptake of system L substrates in astrocytes is needed to confirm the present indications of high affinity and concentrative Na+ independent transport. PMID- 3053995 TI - In memoriam: Eduardo De Robertis 1913-1988. PMID- 3053996 TI - Total hip arthroplasty after renal transplantation. Long-term follow-up study and assessment of metabolic bone status. AB - Twenty-two patients had 36 total hip arthroplasties for painful osteonecrosis of the femoral head. At a mean of 86 months after operation, a complete follow-up evaluation, including physical examination, was obtained in 24 hips in 15 patients. An additional 12 hips in seven patients were followed by telephone interview and radiographic evaluation. Although most patients experienced improved hip function and symptomatic relief from pain as a result of the operation, 10 hips developed heterotopic bone, 5 hips dislocated after operation, 6 hips failed due to aseptic loosening, and 1 hip developed a deep infection, and one patient died due to pulmonary embolism. Neither sex, preoperative steroid dose, nor postoperative mean alternate-day steroid dose could be related to aseptic loosening. However, histologic examination of transilial bone biopsy specimens (7 patients, 13 hips) revealed steroid-induced osteoporosis, by the presence of hyperosteoidosis (increased unmineralized osteoid) and increased bone resorption. Bilateral hip involvement, osteoporosis, and high turnover skeletal remodelling at the cement-bone interface potentially contributed to a failure rate that was higher in this group than that reported for primary hip arthroplasty for other diagnoses. The existence of steroid-induced metabolic bone disease and preexisting renal osteodystrophy may pose a significant threat to the long-term survival of a total hip implant. PMID- 3053997 TI - Transient decrease in IL-2-responsive lymphocytes 24 hours after initiation of continuous IL-2 infusion in cancer patients. AB - Cancer patients were treated with recombinant interleukin-2 (IL-2) in a Phase I clinical trial. Patients were given four repetitive weekly cycles of four days of continuous i.v. IL-2 infusions followed by 3 days of observation. A transient 80% decrease in the number of circulating peripheral blood lymphocytes (PBLs) was noted 24 h after initiation of the IL-2 infusion. The in vitro IL-2 induced proliferative response, and natural killer (NK) activity of the PBLs recovered at this time was only 10-20% of that by the same number of PBLs obtained prior to IL 2 therapy. This effect was transient and rebound increases in circulating lymphocytes expressing high NK and lymphokine-activated killer functions were demonstrated at the end of a 4 day IL-2 infusion. To study further whether the drop in lymphocyte activity observed at initiation of IL-2 therapy was due to activation of a suppressor mechanism, patient PBLs isolated 24 h into the IL-2 infusion were mixed with their pre-IL-2 therapy PBLs at different ratios and assayed in NK and proliferation assays. These mixing experiments did not prove suppression to be the mechanism for the decreased response; rather, the PBLs obtained 24 h into IL-2 therapy merely diluted out the in vitro response of PBLs obtained prior to therapy. Although there was a considerable drop in circulating PBLs 24 h after initiation of each of three subsequent weekly IL-2 treatment cycles, these remaining lymphocytes in the peripheral blood were functional in both NK and IL-2 proliferative assays. These PBLs obtained 24 h into each of the subsequent three cycles showed a progressive increase in their in vitro NK and IL 2-induced proliferative activity, reaching levels two to three times higher than that of pretherapy PBLs by the fourth cycle. Thus, IL-2 caused a transient disappearance of lymphocytes from the circulation at the initiation of each cycle, but lymphocyte function was impaired only in the first IL-2 cycle. These data suggest that resting IL-2 responsive cells initially leave the circulation upon exposure to IL-2, but that such cells become activated and some remain detectable in the circulation when subsequent weekly cycles of IL-2 are given. PMID- 3053998 TI - A comparison of charges for continuous ambulatory peritoneal dialysis and center hemodialysis. AB - Use of continuous ambulatory peritoneal dialysis (CAPD) is increasing, and it is being promoted as a less expensive alternative to center hemodialysis (CHD). The debate over the relative charges for CAPD and CHD cannot generally be answered without considering the relationship between modality selection and patient characteristics. When selection and patient characteristics are accounted for, the difference between annual charges for CAPD and CHD patients is insignificant, in part because of the current payment system. The analysis suggests that patients using CHD may have lower charges than if they were using CAPD; similarly, patients using CAPD may have lower charges than if they were using CHD. Charges during CAPD training are lower than CHD or CAPD charges. Charges during the transition between CHD and CAPD tend to be higher than either CHD or CAPD due to additional hospitalizations. Estimated results suggest that encouraging current CHD patients to transfer to CAPD (or vice versa) may not have the desired effect of reducing charges. PMID- 3053999 TI - A rule for the early detection of chronic changes in cystic fibrosis patient status. AB - A statistical decision-making system has been developed which will predict the clinical status of a patient with cystic fibrosis based on daily self measurements obtained at home. The data for the study were collected from CF patients within 7-12 years of age. Thirty-two participants recorded four daily measurements (weight, vital capacity, breathing rate, and resting pulse) and one weekly measurement (height). In addition to the 4 daily measured values, the clinical status of each patient at his/her most recent previous clinic visit was used as a predictor variable. The measured values were used as the basis for the development of a discriminant rule. The goal of the rule was to determine whether each patient's clinical status was deteriorating, stable, or improving at the time of the most recent set of weekly measurements. Three types of analysis were performed: linear discriminant analysis, quadratic discriminant analysis, and nearest neighbor. Quadratic discriminant analysis provided the best discrimination due to the differences in the covariance matrices among the populations. The rule was able to correctly classify 77% of the 103 cases in the learning set. To further evaluate the rule, both a weighted classification percentage and weighted kappa statistic were calculated for the rule. Bootstrapping was used to predict the performance of the rule on the population with results of 77% correctly classified overall. PMID- 3054000 TI - Estimation of test error rates, disease prevalence and relative risk from misclassified data: a review. AB - We review methods for the analysis of categorical clinical and epidemiological data, in which the observations are subject to misclassification. Under certain conditions, it is possible to estimate error parameters such as sensitivity, specificity, relative risk, or predictive value, even though no definitive classification (gold standard) is available. The parameter estimates are obtained by modelling the data, using maximum likelihood, with or without some constraints. The models recognize that the true classification of an individual is unknown, and so are sometimes referred to as "latent class" models. The latent class approach provides a unified framework for various methods found in a dispersed literature, characterising each by the number of populations or subgroups in the data, and the number of observations made on each individual; the statistical degrees of freedom are implied by the sampling design. Data sets with less than three replicate observations per individual necessarily require constraints for parameter estimation to be possible. Data sets with three or more replicates lead directly to estimates of the misclassification rates, subject to some simple assumptions. Some more complex problems are also discussed, including data where the response variable has more than two levels, sequential and irregular designs and the effects of assumption violations. PMID- 3054001 TI - Intraperitoneal chemotherapy for ovarian carcinoma. PMID- 3054002 TI - Non-Hodgkin's lymphomas in 137 patients aged 70 years or older: a retrospective European Organization for Research and Treatment of Cancer Lymphoma Group Study. AB - The results of a European Organization for Research and Treatment of Cancer (EORTC) retrospective study on non-Hodgkin's lymphoma (NHL) in elderly patients (greater than or equal to 70 years of age) seen in Europe in 1984 are reported. A precodified form was sent to 55 European institutes in order to evaluate the incidence of NHL in the elderly with regard to natural history, treatment-related toxicity, response, and survival. Thirteen institutes participated in the study. One hundred thirty-seven cases of NHL were observed in the elderly during 1984, making up 28% of the total number of NHL seen in those institutes. The median age was 77 years; 21% of the patients had favorable (low-grade) and 73% unfavorable (intermediate- and high-grade) histology, according to the Working Formulation. Stage at presentation was localized (I and II) in 60% and advanced in 37% of the patients. Most of the physicians used standard therapy regimens at reduced doses, from the beginning of the treatment. Sixty patients (44%) underwent a "conservative" treatment (one or two antineoplastic drugs or local field radiotherapy) and 77 (56%) an "aggressive" treatment (polychemotherapy regimens or extended field radiotherapy). Response was similar between the two treatment groups, but severe and lethal toxicity was significantly higher among patients treated with aggressive therapy. Prospective randomized studies are clearly needed to define the optimal treatment in elderly patients with advanced unfavorable NHL. PMID- 3054003 TI - Relapse after interferon alfa-2b therapy for hairy-cell leukemia: analysis of prognostic variables. AB - Sixty-nine patients with hairy-cell leukemia (HCL) were treated with interferon alfa-2b (IFN) in a single-institution study. The dose used was 2 x 10(6) U/m2 self-administered subcutaneously three times weekly, for a planned treatment duration of 12 to 18 months. Of the 68 evaluable patients, the major response rate was 75%, with 13% complete responses (CRs) and 62% partial responses (PRs). An additional eleven patients (16%) had minor responses (MRs). Duration of response was denoted as failure-free survival (FFS), defined as the time from the end of IFN therapy to a need for further antileukemic therapy. Of the 60 responding patients followed after discontinuation of IFN, 27 have relapsed, requiring further therapy. The median actuarial FFS for these 60 patients is 25.4 months. All but five patients are alive, and the actuarial overall survival for the 69 patients is 91% +/- 4% at 4 years from the start of IFN. The best indicators of relapse were the neutrophil alkaline phosphatase (NAP) score and degree of residual bone marrow hairy cells (%HCL) at the completion of therapy. Patients with NAP less than 30 (n = 21) had the best prognosis (median FFS, 30.4 months), while those with NAP greater than or equal to 30 and %HCL less than or equal to 30 (n = 21) or %HCL greater than 30 (n = 16) had intermediate and poor prognoses, respectively (median FFS, 23.5 and 12.4 months) (P = .0005). Fourteen of the relapsing patients are evaluable for response to a second course of IFN, with seven PRs and four MRs. Stratified randomized trials are indicated to determine the role of maintenance therapy for responding patients. PMID- 3054004 TI - How expert physicians would wish to be treated if they had genitourinary cancer. AB - A questionnaire describing six clinical scenarios was mailed to urologists (in Britain, Canada, and the United States) and to radiation and medical oncologists in the United States, who practice genito-urinary (GU) oncology. In each scenario, the surgeon or physician was asked to consider himself as a patient with bladder, prostate, or kidney cancer, and to select his own treatment. Accompanying each scenario were one or two clinical trials for which the physician would be eligible. He was asked to state if he would agree to be randomized in each trial, and if he refused, to state his reasons. We found that (1) there were major differences of opinion about management for each scenario; (2) choice of treatment was influenced more by specialty training or geographic location than by the results of previous clinical trials (which are available to all); (3) British urologists tended to be less aggressive than their North American colleagues, with respect to the use of radical surgery and chemotherapy; (4) acceptance of clinical trials ranged from 3% to 60%; and (5) agreement to clinical trials was quite poor even when they were designed to compare the most popular options for management. This physician surrogate method is a valuable tool in assessment of the degree of consensus amongst expert physicians and in the determination of whether new clinical trials address important areas of controversy. PMID- 3054005 TI - The systemic administration of intravenous melphalan. AB - Melphalan (L-phenylalanine mustard) is a bifunctional alkylating agent that is commonly administered orally to treat a wide variety of malignancies, including cancers of the breast and ovary, as well as multiple myeloma. Although commercially available in Europe and Canada, intravenous (IV) melphalan remains investigational in the United States. The role of IV melphalan in cancer chemotherapy is not well defined, despite its manageable toxicity and higher and more predictable blood levels following IV administration compared with oral administration. In addition, unlike oral melphalan, an extensive phase I evaluation of IV melphalan has not been undertaken. At lower doses (eg, 30 to 70 mg/m2), both as a single agent and in combination, the activity of IV melphalan has been evaluated in only a limited number of diseases. However, striking activity has been observed in previously untreated patients with rhabdomyosarcoma, a disease not generally considered responsive to alkylating agents. When administered at high doses (greater than 140 mg/m2) requiring bone marrow reinfusion, melphalan effects a high response rate (but no improvement in survival) in a variety of nonhematologic tumor types, including resistant tumors such as melanoma and colon carcinoma. In contrast, in poor-prognosis patients with non-Hodgkin's lymphoma, Hodgkin's disease, multiple myeloma, or neuroblastoma, high-dose melphalan-containing regimens have yielded both high response rates and improved survival, despite considerable toxicity. Additional clinical trials will be necessary to define the spectrum of activity of lower doses of IV melphalan and to define subgroups of patients most likely to benefit from high-dose melphalan. PMID- 3054006 TI - Extension of remission in acute nonlymphocytic leukemia. PMID- 3054007 TI - Treatment of multiple myeloma: an Argentine Group for the Treatment of Acute Leukemia Study. PMID- 3054008 TI - Embryonic divergence of oligodendrocyte and astrocyte lineages in developing rat cerebrum. AB - Oligodendrocyte and astrocyte lineages were traced in rat forebrain sections using single- and double-label immunoperoxidase and indirect immunofluorescent techniques. Antibodies were directed against antigenic markers, the expressions of which overlapped in time: GD3 ganglioside in immature neuroectodermal cells; vimentin in radial glia; glial fibrillary acidic protein (GFAP) in astrocytes; and carbonic anhydrase (CA) and galactocerebroside (GC) in oligodendrocytes. A histochemical stain for iron was also used as a marker of oligodendrocytes. Small cells of the subventricular zone (SVZ) were stained with anti-GD3 but not with the other antibodies. By 16 d of gestation (E16), the SVZ generated large, round cells and thick, process-bearing cells that were GD3+/CA+/iron+. These cells then appeared in the cingulum and, with time, increased in numbers and extended thick processes as they filled the subcortical white matter. These cells eventually lost their reactivity to anti-GD3 but became GC+/CA+ with processes extending to myelin sheaths. At E15 radial glia were stained with the anti-vimentin antibody but were negative for GFAP. At birth, only the vimentin+ radial glia midline between the 2 ventricles were GFAP+, but with time more vimentin+ cells became GFAP+. By 7 d of postnatal age all the vimentin+ cells were GFAP+ and had converged predominately on the cingulum. With time these cells condensed and took on characteristic shapes of astrocytes. The embryonic separation of the oligodendrocyte and the astrocyte lineage is supported by four pieces of evidence: (1) GD3+ cells were double labeled with anti-CA, and then went on to become GC+; (2) vimentin+ and GFAP+ cells were not also GD3+; (3) ultrastructural localization of anti-GD3 was confined to cells with characteristics consistent with developing oligodendrocytes; and (4) the shapes of GD3+, CA+, GC+, or iron+ cells did not resemble those of the vimentin+ or GFAP+ cells. PMID- 3054009 TI - Controlled outgrowth of dissociated neurons on patterned substrates. AB - The cytoarchitecture of nervous tissue is lost during the dissociation procedures used to form primary cell cultures. As a first step toward reestablishing an ordered arrangement of these cells in vitro, we developed a set of procedures for patterning the outgrowth of cells cultured on 2-dimensional substrates. These procedures used a combination of surface chemistry and photolithographic techniques. The adhesive properties of either silicon or silicon dioxide (quartz) surfaces were controlled by covalently binding small organic molecules to the surface with silane coupling agents. The attachment and growth of either embryonic mouse spinal cells or perinatal rat cerebellar cells were found to be promoted by binding certain amine derivatives to the surface. In particular, cells grown on surfaces bound with diamines and triamines, but not with monoamines, formed cultures whose morphology was similar to that of cells cultured on conventional substrates, i.e., glass coated with poly(D-lysine). The attachment of cells to a substrate was inhibited by binding alkane chains (e.g., n-tetradecane) to the surface and plating the cells in media containing 5-10% (vol/vol) serum. Patterns of selected adhesivity were formed using photochemical resist materials and lithographic masking techniques compatible with the silane chemistry. Cultures of either spinal cord cells or cerebellar cells could be confined to square regions on the scale of 50 micron. Cerebellar cells could be confined to grow on lines with widths less than 10 micron. This width is comparable to the diameter of granule cell somata. The patterned growth of cerebellar cells was maintained up to 12 d in vitro. Over this time period the granule cells were observed to develop electrical excitability and immunoreactivity for neuron-specific enolase. Purkinje neurons also developed electrical excitability when grown on the chemically modified surfaces. Immunochemical reactivity of the patterned cultures for glial fibrillary acid protein (GFAP) showed that glia are patterned along with the associated granule cells. Interestingly, the GFAP-positive glia that proliferated on surfaces bound with amine derivatives attained primarily a tile-shaped, fibroblast-like morphology, while those proliferating on glass coated with poly(D-lysine) developed primarily a spindle-shaped, process-bearing morphology. Granule cells preferentially associated with the spindle-shaped glia. PMID- 3054011 TI - A double-blind placebo-controlled trial of perioperative prophylactic antibiotics for elective neurosurgery. AB - In this study, 417 patients undergoing "clean" elective neurosurgical operative procedures were randomized to receive a broad-spectrum antibiotic (piperacillin) or placebo given as three perioperative doses, each 6 hours apart. Randomization was carried out by hospital pharmacists, and the investigators remained blinded until the end of the study. Twenty cases were excluded from analysis because either an unforeseen second operation was performed or antibiotic therapy was initiated within 30 days after surgery to treat infection or the risk of infection. Twelve of the 205 patients treated with placebo developed postoperative wound sepsis, and four of the 192 piperacillin-treated patients developed wound sepsis--a statistically significant difference (p less than 0.05, Fisher's exact test). Piperacillin thus appeared to reduce the incidence of neurosurgical wound infection in this study. PMID- 3054012 TI - Current reporting of responsiveness in acute cerebral disorders. A survey of the neurosurgical literature. AB - One hundred sixty-six papers published in seven neurosurgical journals from 1983 through 1985 have been surveyed to determine the methods used for assessment of overall patient responsiveness in acute cerebral disorders (coma grading). Fifty one different coma scales or modifications were found. The Glasgow Coma Scale (GCS) sum score (that is, the sum of the scores of the individual eye, verbal, and motor scales) dominated (54%), and was used in 73 (76%) of 96 of the head injury studies; in 56 (77%) of these 73 studies it was the single method of grading neurological status. The GCS sum score was used in 16 (23%) of 70 studies in patients with other etiologies. The Hunt and Hess scale was used in 26 (57%) of 46 reports of patients with subarachnoid hemorrhage. In 31 (55%) of the 56 studies of head injuries using the GCS alone, it was not obvious if the 12- or 13 grade scale was used. In 13 studies (23%) no reference to methodological investigations was made. In 44 papers (79%) the handling of untestable features, such as intubation or swollen eyes, was not reported. In the 56 studies using the GCS alone, coma was defined in many different ways and in 22 studies the definition of coma was not specified. In 63% of reports, the GCS sum score scale was combined in one to five groups of scores and this was done in 32 different ways. No information was available to describe the procedure of data aggregation or the reliability of the 13-grade GCS sum score. The lack of standardization makes it unnecessarily difficult to perform valid comparisons between different series of patients. Since the GCS sum score is the most widely used scale, it is suggested that the reporting of the GCS sum score should be standardized regarding pseudoscoring, coma definition, and use of combined scores. Further studies on the reliability of the GCS sum score are needed. PMID- 3054010 TI - Effect of nimodipine on the outcome of patients after aneurysmal subarachnoid hemorrhage and surgery. AB - The effect of intravenous nimodipine on the incidence of mortality and delayed ischemic neurological deficits of patients after aneurysmal subarachnoid hemorrhage (SAH) and surgery was studied in a prospective double-blind placebo controlled trial. Upon admission, all of the patients were in Grades I to III according to the classification of Hunt and Hess. Of the 213 patients enrolled in the study, 58 underwent early surgery (within 72 hours after the bleed: Days 0 to 3), 69 were operated on subacutely (between Days 4 and 7), and 74 had late surgery (on Day 8 or later). Eleven patients died before surgery was undertaken and one was not scheduled for operation. Administration of the drug was started immediately after the radiological diagnosis of a ruptured aneurysm had been made. The dose of nimodipine or matching placebo was 0.5 micrograms/kg/min via continuous intravenous infusion for 7 to 10 days after the SAH and, if the patient was operated on late, for 2 to 3 days after the operation as well. After intravenous treatment, oral administration of nimodipine or placebo was continued for up to 21 days after SAH in a dose of 60 mg every 4 hours. Nimodipine treatment was associated with a significant decrease in mortality rate (p = 0.03) in the early and subacute surgery groups. In the total series the number of deaths due to delayed ischemic deterioration was significantly lower in the nimodipine group than in the placebo group (p = 0.01). PMID- 3054013 TI - A comparison of the Glasgow Coma Scale and the Reaction Level Scale (RLS85). AB - The Glasgow Coma Scale (GCS) and the Reaction Level Scale (RLS85) were compared for rating neurosurgical patients in regard to ranking order of deficit severity, interobserver variability, and coverage for relevant factors. Four physicians, four registered nurses, and four assistant nurses performed 72 pairwise ratings on 47 neurosurgical patients. The rank correlation between the GCS sum score and the RLS85 was -0.94, suggesting the same ranking order of severity and indicating that the underlying concepts of somnolence, delirium, and motor responses in coma are evaluated in the same way. By the sign test, the RLS85 was shown to have better interobserver agreement than the GCS sum score and the eye-motor-verbal (EMV) profile. The interobserver grading disagreements in both scales were distributed over the entire range of responsiveness, and for the GCS sum score they were slanted to combined segments 9 to 15. The RLS85 showed full coverage of relevant factors, while 43 (60%) of the 72 test occasions in the GCS sum score and the EMV profiles showed untestable features, most often because of patient intubation. The pseudoscore (that is, the choice of value given to untestable features) affects interobserver agreement as well as the estimated overall patient responsiveness in the GCS sum score. Assessment by the order of applying the scales showed a significant effect on the GCS eye-opening scale (p = 0.01) and the GCS sum score (p = 0.03), indicating a sensitivity to environmental stimuli unrelated to the patient's status. This study demonstrates that basically the same information as that found in the separate eye, motor, and verbal scales of the GCS can be combined directly into the RLS85, which has better interobserver agreement and better coverage than the GCS sum score. PMID- 3054014 TI - Isolation and in vitro growth of glioma-infiltrating lymphocytes, and an analysis of their surface phenotypes. AB - The present investigation was conducted in order to examine the feasibility of isolating and growing glioma-infiltrating lymphocytes in vitro as possible effector cells for use in new adoptive immunotherapy. Eight surgical specimens obtained from patients with malignant astrocytomas were treated by enzyme dispersion; the cells were separated on a density gradient and grown in the presence of human recombinant interleukin-2. The cultured lymphocytes were tested for cell-surface markers by using monoclonal antibodies in a flow cytometric analysis. In all cases the glioma-derived lymphocytes were grown in culture for several weeks with substantial increases in cell numbers (at least 5 X 10(8) cells). The mature T cell population (CD3, 89%) was found to have an increased proportion of the cytotoxic/suppressor phenotype CD8 (55%) as compared to peripheral blood lymphocytes (PBL's). Eighty-six percent of the cultivated lymphocytes expressed HLA-DR. The IL-2 receptor was predominantly expressed on the helper subset (CD4-positive). Otherwise, anti-CD16, which specifically reacts with natural killer (NK) cells, did not stain significantly more of the cultured gliomaderived lymphocytes compared with lymphocyte-activated PBL's. These results corroborate the observations made with conventional immunohistochemical examination. It has been demonstrated that T lymphocytes isolated from human cancers are enriched for specific reactivity to their autochthonous tumor cells. These experiments support the possible use of glioma-infiltrating lymphocytes as a new treatment for patients with malignant glioma. PMID- 3054015 TI - Creutzfeldt-Jakob disease probably acquired from a cadaveric dura mater graft. Case report. AB - A case of Creutzfeldt-Jakob disease (CJD) is reported in a 28-year-old woman who had received a cadaveric dural graft 19 months earlier after resection of a cholesteatoma. The circumstances of the case point to the graft as the most likely source of the disease. Cadaveric dura should be added to the list of materials that may transmit CJD, and it must be very carefully screened if it is used at all for grafting. Autologous tissue should be considered whenever possible. PMID- 3054016 TI - Nimodipine treatment in poor-grade aneurysm patients. PMID- 3054017 TI - Calculation of portal contribution to hepatic blood flow with 99mTc microcolloids. A noninvasive method to diagnose liver graft rejection. AB - The portal contribution (PC) to hepatic blood flow was calculated in 13 liver graft patients and 13 normal volunteers. The method is based on the quantification and normalization of the liver and spleen activity after the administration of 7 mCi (259 MBq) of 99mTc microcolloid. Forty examinations were performed in liver grafts and 13 in normal subjects. The PC was significantly higher in normal native liver (64.0 +/- 3.0%) than in functioning grafts (58.8 +/ 3.1%). In acutely rejecting patients, PC was significantly lower (52.4 +/- 2.0%) than in functioning grafts and similar to that observed in cholangitis (53.5 +/- 0.7%). The PC increases again once rejection has resolved (57.3 +/- 2.6%). During hepatitis post-transplant PC values (59.7 +/- 3.4%) were similar to those observed in functioning grafts. Overall, PC values over 55% are very unlikely to be due to rejection. PMID- 3054018 TI - Budgets: an underused resource. AB - Though budgeting is an activity that management readily participates in, the budget often ends up being underused. The author discusses how budgeting activities can be used in revenue allocation, program implementation, planning, coordination, profitability, designation of responsibility, and measurement reviews. PMID- 3054019 TI - Taurine in development. AB - Taurine is a ubiquitous dietary constituent of most mammals and is present in especially high concentrations in the tissues of developing mammals. Research to date indicates that taurine plays an important role in the development of the nervous system and the process of migration in particular. It is speculated that taurine uptake and release, in conjunction with glutamate uptake and release, may represent one form of communication between neurons and glial cells. The need of taurine by the body is emphasized by the ability of the kidney to curtail taurine excretion to conserve taurine in the face of a low dietary taurine intake. The evidence for a special role of taurine in development is considered and discussed. PMID- 3054020 TI - The effect of past and current dietary Zn intake on Zn absorption and endogenous excretion in the rat. AB - Effects of previous dietary Zn (or body Zn stores) and current dietary Zn intake on absorption and endogenous excretion of Zn were studied by using radioisotope dilution. Rats were fed diets containing 1.5, 12.6 or 50.3 mg Zn/kg for 19 d (dietary period I). Total body Zn in the three groups was 1870 +/- 340, 3953 +/- 698 and 4126 +/- 844 micrograms Zn/rat. Each group was divided into four subgroups fed 3.6, 12.6, 20.5 or 50.3 mg Zn/kg diet for 3 wk (dietary period II). Rats were injected intramuscularly with 65Zn after 7 d of dietary period II. True absorption and endogenous excretion were calculated by isotope dilution. Zinc intake, urinary and fecal excretion, balance and percent Zn absorption were significantly affected only by dietary Zn in dietary period II (P less than 0.01). Endogenous excretion was affected by both past dietary Zn deficiency (body Zn stores) and by dietary Zn in dietary period II (P = 0.0001). Total body Zn at the end of the experiment was significantly affected by both periods of dietary treatment (P less than 0.001), but total body Zn concentration was affected only by the final dietary treatment (P less than 0.05). These results show that Zn absorption is affected by the current diet, but that turnover of Zn (endogenous excretion) is regulated by both current Zn intake and past Zn intake, probably through an effect on body Zn stores. PMID- 3054021 TI - Glucose kinetics in protein depletion--effect of glucose infusion in the fasted rat. AB - A primed constant infusion of [6-3H]glucose was used to determine glucose turnover in rats fasted for 48 h that had previously been pair-fed isoenergetic 4% and 14.4% protein diets for 6 wk. The protein-depleted rats had a low body weight and lower plasma glucose and plasma insulin concentrations compared with the 14.4% protein pair-fed control animals. The rate of appearance of glucose in plasma in the protein-depleted rats was significantly less than that in the pair fed controls. During an intravenous infusion of unlabeled glucose (38.8 mumol.kg 1.min-1) plasma glucose concentrations and glucose appearance increased in both groups. Plasma insulin concentration remained lower in the 4% protein-fed group though the percentage of increase above basal was similar in both groups during glucose infusion. The clearance of glucose from the blood and suppression of endogenous glucose production during the exogenous glucose infusion were similar in both control and protein-depleted rats. Rats fed a 14.4% protein diet ad libitum consumed more diet than the pair-fed groups and their body weights were greater. However, fasting plasma glucose and insulin, concentrations were similar to the 14.4% protein pair-fed animals. This indicates that the observed changes in glucose metabolism in the 4% protein-fed animals were due to a reduced dietary ratio of protein to carbohydrate rather than reduced energy intake. PMID- 3054022 TI - Cholesterol levels and eicosanoid production in rats fed phosphatidylinositol or soybean lecithin. AB - Male young rats were fed 8% corn oil diets supplemented either with 2% phosphatidylinositol (PI) from safflower seeds or soybean lecithin (SL) for 22 days. Other groups of rats were fed 10% corn oil diets with or without (control) 0.3% inositol (IN, equivalent to the inositol moiety of the PI diet). The plasma cholesterol level was low in the SL group whereas liver triglyceride was low in all supplemented groups. The aortic production of prostacyclin tended to be high in rats fed the control diet and low in rats fed the SL diet, the PI and IN groups being intermediate. The concentration of plasma thromboxane B2 was comparable among various groups. In plasma and liver phosphatidylcholine, the ratio of arachidonate/linoleate was low in rats fed SL and high in rats fed PI or IN diets. The results indicate that, in addition to SL, the inositol moiety of PI may have a significant role in the regulation of lipid metabolism. PMID- 3054023 TI - Early diagnosis of acoustic neuroma. PMID- 3054024 TI - Sicca syndrome due to amyloidosis. PMID- 3054025 TI - Cat scratch fever presenting as a submental swelling. PMID- 3054026 TI - The autogenous dermal graft in temporomandibular joint disc surgery. AB - Repair or replacement of the disc in 58 patients (64 joints) with temporomandibular joint internal derangement was done using an autogenous dermal graft. Long-term follow-up of 3 to 8 years revealed successful elimination of symptoms and restoration of mandibular function in 51 patients (87.9%). The autogenous dermal graft, rather than alloplastic materials, may be the procedure of choice when repair or replacement of the temporomandibular joint disc is indicated. PMID- 3054027 TI - Porous hydroxylapatite granules and blocks as alveolar ridge augmentation materials: a preliminary report. AB - Porous hydroxylapatite (PHA) blocks and granules were used in the augmentation of 30 maxillary and mandibular ridges in 28 patients. The postoperative evaluation period was 2 years in all cases. The patients were evaluated clinically and radiographically, and by patient questionnaires. An increased incidence of dehiscence was noted with the blocks as compared with the granules. Overall prosthodontic assessment showed 95% improvement among the granule cases when compared with the preoperative ridge, and 88% improvement noted among the block cases. Patient rating of their general satisfaction with their dentures showed 82% improvement in the granule patients, and 55% in the block cases. Radiographically, the granules showed an average decrease of 8%. The results of this study show that PHA granules can be used as satisfactory alveolar ridge augmentation material, while the blocks show an increased number of complications and should be used only in very selected cases. PMID- 3054028 TI - Orbital infections: clinical and radiographic diagnosis and surgical treatment. AB - Orbital infections are uncommon sequelae of sinusitis, odontogenic infections, or orbital trauma and may have devastating consequences if they are unrecognized or if aggressive treatment is delayed. The authors present a systematic classification of orbital infections, and discuss the pathogenesis and treatment of orbital abscesses. PMID- 3054029 TI - [Ultrasonography of mucoceles of paranasal sinuses--a secondary report]. PMID- 3054030 TI - [A study on endothelial cell growth factors from the thyroid anaplastic cell carcinoma using human umbilical vein-endothelial cells]. PMID- 3054031 TI - [Three cases of neurinoma arising from the vagus nerve in the neck]. PMID- 3054032 TI - Funeral customs in historical context. PMID- 3054033 TI - Chronic renal failure in infants and children. AB - Chronic renal failure is an uncommon problem for pediatricians, but early recognition is important for maximizing growth and minimizing complications. Marked strides have been made in understanding and treating renal osteodystrophy. Recombinant erythropoietin holds the promise of reversing the anemia associated with renal insufficiency. Dialysis remains an important therapy for sustaining these children, and transplantation offers realistic hope for a functioning kidney. PMID- 3054034 TI - Renal injury in the asphyxiated newborn infant: relationship to neurologic outcome. AB - The relationship of renal and central nervous system injury was prospectively evaluated in 120 asphyxiated infants. Renal evaluation findings were considered abnormal if there was oliguria (urine output less than 1 ml/kg/hr), which was designated transient if present in the first 24 hours only and persistent if present for at least 36 hours, or if the urinary beta 2-microglobulin concentration from first-void urine was elevated: (1) Thirteen infants had persistent oliguria; the urinary beta 2-microglobulin level was elevated in all. The six term infants had clinical signs consistent with hypoxic-ischemic encephalopathy (HIE); all six had ultrasonographic abnormalities. The outcome was poor (i.e., death or long-term neurologic deficits) in five of six infants. The seven preterm infants with persistent oliguria had clinical evidence of HIE, and three infants had intraventricular hemorrhage; all seven infants died. (2) Fifteen infants had transient oliguria (beta 2-microglobulin level was elevated in eight infants). Two of the eight term infants had evidence for HIE; the cranial ultrasound scan was normal in all. At follow-up, seven term infants are normal and one is abnormal. Six of the seven preterm infants with transient oliguria had clinical evidence of HIE; three infants had intraventricular hemorrhage. Three infants died, and the four survivors are normal at follow-up. (3) Ninety-two infants had normal urine output. Of the 22 term infants, two developed signs of HIE, and the ultrasound scan was abnormal in three infants. Of the 70 preterm infants, eight (11%) had clinical signs consistent with HIE, the ultrasound scan was abnormal in 20 of 64 (31%) infants scanned, and 14 (20%) infants died. Most of the followed infants are normal. Thus oliguria was significantly associated with clinical signs of HIE, including seizures, death (specifically in the premature infant), and long-term neurologic deficits. These data suggest that oliguria in the perinatal period is a sensitive indicator of infants at risk for long-term neurologic deficits. PMID- 3054035 TI - Therapy for shigellosis. I. Randomized, double-blind trial of nalidixic acid in childhood shigellosis. AB - We compared nalidixic acid, 55 mg/kg/day, with ampicillin, 100 mg/kg/day, both given orally for 5 days, in the treatment of children with dysentery caused by shigellosis. All patients entered into the study had illness of less than 72 hours' duration and no prior allopathic drug therapy. Treatment was randomized and administered in double-blind fashion. Patients initially treated with ampicillin who were infected with a Shigella strain resistant to ampicillin were considered as a separate group (ampicillin-R). All isolates were susceptible to nalidixic acid. Similar percentages of patients treated with nalidixic acid (26/32, 81%) and with ampicillin (17/22, 77%) were clinically cured by the end of therapy; the rate in ampicillin-R (3/14, 21%) patients was significantly lower (p less than 0.001). Stool frequency in patients treated with nalidixic acid was significantly less than for ampicillin-treated or ampicillin-R patients during the final 3 study days. All patients treated with nalidixic acid and ampicillin had Shigella eradicated from their stool by day 3, compared with 77% (10/13) of ampicillin-R patients (p less than 0.05, ampicillin-R vs nalidixic acid or ampicillin). We conclude that nalidixic acid is an effective alternative to ampicillin in the treatment of shigellosis caused by nalidixic acid-susceptible strains. PMID- 3054036 TI - FDA reorganization. PMID- 3054038 TI - Bowel habit from birth to old age. PMID- 3054037 TI - Kwashiorkor and aflatoxins. PMID- 3054039 TI - Propylthiouracil hepatotoxicity: two pediatric cases and review of the literature. AB - We have observed isolated hepatotoxicity in two children treated with propylthiouracil (PTU) for hyperthyroidism. Neither patient had risk factors for or clinical evidence of preexisting liver disease. In one patient the drug was promptly discontinued when signs of liver disease were noted. This patient quickly recovered. The second patient continued to receive PTU for several days after developing symptoms. Her illness progressed to fulminant hepatic failure with encephalopathy, and she died. These are the third and fourth pediatric cases reported, and there have been 10 cases reported in adults in the English language literature. Thirteen of the 14 patients are female. The literature regarding all these patients is reviewed. Propylthiouracil may cause lethal hepatic damage. This drug should be discontinued immediately if signs or symptoms of hepatic injury are detected. PMID- 3054040 TI - Ascites complicating ventriculoperitoneal shunts. AB - Ventriculoperitoneal shunts are currently a standard therapy for obstructive hydrocephalus. These shunts are associated with a variety of abdominal complications, one of which is the development of ascites. We report an 11-year old girl with a ventriculoperitoneal shunt in whom a low-grade peritoneal infection presented with ascites. This case demonstrates the importance of diagnostic paracenteses, appropriate antibiotic therapy and the potential need to establish an alternative route for cerebrospinal fluid diversion in patients with ventriculoperitoneal shunts and ascites. PMID- 3054041 TI - Enema reduction of intussusception by hydrostatic pressure under ultrasound guidance: a report of 377 cases. AB - Three hundred seventy-seven pediatric intussusception patients were treated by normal saline hydrostatic enema under ultrasound guidance from October 1985 to April 1987. Before reduction, the rate of correct diagnosis by ultrasonography was 100%, and the rate of successful reduction was 95.5% in this group. With this technique there is no risk of x-ray exposure to the patient. The definite criteria for reduction are discussed. Clear echogram was shown during reduction and the ileo-ileo-coli intussusception can be diagnosed. Intestinal perforation, if any, can be accurately recognized at once. This technique is believed to be one of the most promising methods in nonoperative treatment of pediatric intussusception. PMID- 3054042 TI - Evaluation of immune status in pediatric recipients of hepatic orthografts. AB - Survival in children following hepatic transplantation is better than in adults. We evaluated the immunologic status of both pediatric and adult patients prior to and following transplantation, to determine if there were differences in immunologic status that might contribute to this phenomenon. Studies included determination of mononuclear cell subsets, including T cells, T helper cells, T suppressor cells, and monocytes. In addition, we evaluated immune function by studying in vitro immunoglobulin (Ig) synthesis. Following transplantation, the T8 (cytotoxic/suppressor) cells in pediatric patients did not decline, whereas in adults they did (P less than .05). In vitro Ig synthesis in both adult and pediatric patients declined following transplantation. By 4 weeks, the adult group had begun to recover, whereas in the pediatric group there was still significant suppression (P less than .05). These studies suggest that there is decreased immune function in children who are recipients of hepatic allografts. Such findings could contribute to better outcome of hepatic allografting in children. PMID- 3054043 TI - Asthma update. Part I. Mechanisms, pathophysiology, and diagnosis. PMID- 3054044 TI - Asthma update. Part II. Treatment. PMID- 3054045 TI - Effect of periodontal therapy on alveolar bone as measured by subtraction radiography. AB - Changes in the periodontal alveolar bone are often evaluated by comparing a series of radiographs taken over time. This investigation used a technique that allowed the image registration to be geometrically standardized each time a radiograph was taken. Radiographs of 24 patients from an ongoing double-blind, clinical study using metronidazole were obtained: (1) before any treatment, (2) at the completion of scaling and root planing and surgery (when performed) and (3) during the maintenance phase. One hundred six (106) paired comparisons were analyzed by subtraction radiography using a computerized system. Of these, 95 (89%) exhibited a minimal degree of geometric distortion and could be successfully substracted. Most areas (67%) showed no change in bone structure following periodontal treatment. Bone gain was noted in 12% of the sites examined, while bone loss was seen in 21% of the sites. This bone loss was statistically associated with sites that had received some form of surgical treatment. PMID- 3054046 TI - Lipopolysaccharide-stimulated PGE2 release from human monocytes. Comparison of lipopolysaccharides prepared from suspected periodontal pathogens. AB - Lipopolysaccharides (LPS) prepared from the suspected periodontal pathogens Actinobacillus actinomycetemcomitans (A. a.), Bacteroides gingivalis, B. intermedius and Wolinella recta were compared to Salmonella typhimurium LPS for their capacity to stimulate prostaglandin E2 (PGE2) release from human monocytes. Counterflow isolated monocytes were cultured with control medium or media containing 10 micrograms/ml LPS. Media were then exchanged every 24 hours for a total of 72 hours. Salmonella and Wolinella LPS preparations demonstrated seven fold greater PGE2 release than B. gingivalis and two-fold greater than A. a. and B. intermedius. PGE2 release was found to decrease over time with all LPS preparations except Wolinella. The potency of the LPS preparations is tentatively ranked as follows: Wolinella greater than or equal to Salmonella greater than A. a. greater than B. intermedius greater than or equal to B. gingivalis. These findings demonstrate that LPS preparations from suspected periodontal pathogens are capable of stimulating PGE2 release from human monocytes. The high potency and prolonged stimulation of PGE2 release with Wolinella LPS suggests unusual toxic properties that may exert a greater influence in the pathogenesis of destructive periodontal diseases. PMID- 3054047 TI - Use of antimicrobial containing acrylic strips in the treatment of chronic periodontal disease. A three month follow-up study. AB - Local antimicrobial therapy has been considered for use in the treatment of chronic periodontal disease. This study evaluated chlorhexidine, metronidazole, and tetracycline delivered into periodontal pockets in an acrylic resin vehicle and compared the results with root planed and untreated sites over a three-month follow-up period. One site per patient where pocketing greater than or equal to 6 mm associated with a single rooted tooth was randomly allocated to one of the five possible regimens. Baseline and follow-up measurements included probing depth, loss of attachment, bleeding on probing, crevicular fluid flow, and dark field microscopy of a subgingival plaque sample. Intratreatment evaluations revealed no significant changes in any parameter for untreated sites. Significant improvements in many parameters occurred with all four therapies although the magnitude and duration were greater in metronidazole and root planing groups. The more important intertreatment comparisons indicated that most treatments produced significant benefits compared with the control group; however, again these were greater with metronidazole and root planing. Furthermore, significantly greater effects were noted for metronidazole and root planing compared with tetracycline and more particularly chlorhexidine. It is concluded that some locally delivered antimicrobials alone may be useful in the treatment of chronic periodontal disease. However, at this time local antimicrobial therapy should be considered as adjunctive to conventional debridement techniques. PMID- 3054049 TI - Radiographic evaluation of the effect of initial periodontal therapy on thickness of the maxillary sinus mucosa. AB - The thickness of the radiographic image of the maxillary sinus mucosa on intraoral radiographs was evaluated in 13 patients with advanced periodontal disease, prior to and 12 months following initial periodontal therapy. Before treatment, a relationship was observed between the thickness of the sinus mucosa and the mean probing depths of the teeth in the involved sextant. As many as 79% of the available sextants showed swelling of the mucosa prior to periodontal therapy, compared to only 17% after treatment. This report supports previous studies indicating that advanced periodontal disease may cause swelling of the maxillary sinus mucosa and that periodontal therapy will significantly reduce such swelling. PMID- 3054048 TI - Clinical evaluation of the use of citric acid and autologous fibronectin in periodontal surgery. AB - This study evaluated the effects of citric acid demineralization and autologous fibronectin application in association with a modified Widman flap in the treatment of periodontitis. The study population comprised 29 patients under treatment for moderate to advanced periodontitis who reached the one-year posttherapy evaluation. After thorough scaling and root planing, a split mouth design was used in which two quadrants were treated by modified Widman flap alone, and the other two randomly assigned quadrants were treated by modified Widman flap combined with citric acid demineralization and autologous fibronectin application. Fibronectin, which had previously been isolated from the patient's own plasma, was applied with a tuberculin syringe on the citric acid demineralized root surfaces and the inner aspect of the flap. After suturing provided good flap adaptation, additional fibronectin was again applied under the flap and external pressure was applied. Patients were clinically evaluated at baseline and at one year. Statistical evaluation of the data using paired t test and Chi-square analysis indicated that both approaches, modified Widman flap alone or in combination with citric acid and fibronectin, significantly reduced probing pocket depth and increased clinical attachment. However, the changes achieved with citric acid and fibronectin were statistically greater than those obtained with the flap alone. Furthermore, the number of sites gaining 2 mm or more of clinical attachment were significantly increased. The results suggest that the use of citric acid and fibronectin holds promise in promoting reattachment after periodontal therapy. PMID- 3054050 TI - Pemphigus vulgaris presenting as a gingival lesion. A case report. AB - A 26-year-old hispanic male presented with a nonspecific gingival lesion initially presumed to be of infectious etiology. During continued follow-up over several weeks, the lesion evolved into one typical of desquamative gingivitis. Direct immunofluorescent testing and routine histopathology resulted in a diagnosis of pemphigus vulgaris, which was confirmed by indirect immunofluorescence. Although it is unusual for pemphigus vulgaris to present with the gingiva as the sole primary site of involvement, this case serves to enhance our awareness of the gingiva as a site at which systemic disease can be manifested. PMID- 3054052 TI - Use of collagen shields as a surgical adjunct. AB - We used porcine collagen shields combined with tobramycin, gentamicin, pilocarpine, dexamethasone, and flurbiprofen sodium in 67 cases of penetrating keratoplasty and 55 cases of cataract extraction. No adverse effects were noted from the combined use of the shields with the drug. The devices proved beneficial for protection, lubrication, enhancement of epithelialization, and drug delivery. PMID- 3054051 TI - A kinetic study of chlorpromazine on the hyperglycemic response in rats. II. Effect of chlorpromazine on plasma glucose. AB - Kinetics of the pharmacologic effect of chlorpromazine was investigated in intact fed rats. After i.v. bolus administration of chlorpromazine (0.5, 2, 4 mg/kg), the time courses of plasma glucose, insulin, adrenaline and noradrenaline levels as well as serum and brain concentrations of the drug were determined. Since the hyperglycemic effect of chlorpromazine is known to be attributable to the endogenously released catecholamines, the effects of adrenaline and noradrenaline on the plasma glucose and insulin regulation system were also determined. The plasma glucose regulation system in rats was investigated by an i.v. glucose tolerance test. From the data obtained, a pharmacokinetic (PK)-pharmacodynamic (PD) model as well as the plasma glucose regulation model was constructed. The hyperglycemic effects of catecholamines during and after i.v. infusion were reasonably well correlated with the plasma concentrations of catecholamines using the PK-PD model. Since a quantitative relationship between plasma concentrations of catecholamines and the brain concentration of chlorpromazine was established in the previous report, the time course of hyperglycemic effect of chlorpromazine was analyzed. The result indicated that the hyperglycemic effect can be described quantitatively by a simple PK-PD model with plasma glucose regulation system, using brain concentrations of chlorpromazine in rats. PMID- 3054053 TI - Effect of penetrating keratoplasty using grafts of various sizes on keratoconic myopia and astigmatism. AB - The records of 72 consecutive keratoconic eyes undergoing penetrating keratoplasty were reviewed for changes in myopia and astigmatism. Ages of the patients averaged 32.7 years. All sutures were removed after three months. Follow up average was 40.2 months. Results showed an average decrease in myopia of 6.63 diopters (D) in 60 eyes (82.86%) and an average increase in myopia of 1.88 D in 12 of 70 eyes (17.14%). The decrease/increase in myopia and postoperative astigmatism was compared for grafts equal to the opening, grafts smaller than the opening, and grafts larger than the opening. The largest average decrease in myopia was 13.86 D (range 6.63 to 20.00), which occurred when a graft smaller than the opening was used (P less than .01). This group also showed the least postoperative astigmatism (2.82 D) (P less than or equal to .01). From this study, it appears that the use of a graft 0.25 mm smaller than the trephine opening in the host (i.e., 7.50 mm graft/7.75 mm opening) for penetrating keratoplasty in keratoconus is justified. A prospective study is now in progress. PMID- 3054054 TI - Practice styles and preferences of ASCRS members--1987 survey. AB - A survey of the practice styles and preferences of 1987 ASCRS members was taken in the summer of 1987. A total of 3,809 surveys were mailed in July and 37% were returned. This response rate was 8% higher than the previous year. No financial incentive to return the survey was given. The responses were analyzed using DBase III. There were four main profile questions that dealt with practice style, volume of cataract surgery, geographical location, and age of the ophthalmologist. One section compared how those who prefer phacoemulsification answered questions on phacoemulsification with how those who prefer planned extracapsular cataract extraction answered the same questions. Ophthalmologists were questioned on surgery techniques, intraocular lens style preferences, radial keratotomy, laser use, and assorted miscellaneous items. Whenever possible, the results of questions from the previous two years were compared. PMID- 3054055 TI - Fluorophotometric evaluation of the blood-ocular barrier function following cataract surgery and intraocular lens implantation. AB - Ocular fluorophotometry is a sensitive method for quantitative, in vivo evaluation of the blood-ocular barrier function. This paper reviews studies that have used this technique to evaluate the effects of devices or techniques used in cataract and implant surgery. PMID- 3054056 TI - Cataract surgical suture marking device for the intraoperative control of astigmatism. AB - A new device for the intraoperative monitoring of astigmatism is described. Use of the instrument enables the surgeon to position the closing sutures precisely and minimize astigmatic errors often induced at closure. PMID- 3054057 TI - Nitrofurantoin-stimulated proteolysis in human erythrocytes: a novel index of toxic insult by nitroaromatics. AB - Nitrofurantoin is an antimicrobial agent that causes nonimmune hemolytic anemia in susceptible populations and produces oxidant stress and cellular damage by mechanisms that differ from those associated with oxidants such as phenylhydrazine, which has been shown to stimulate proteolysis in red cells (Goldberg and Boches, 1982). Thus a study of the effects of nitrofurantoin on proteolysis in normal human red cells and red cell hemolysate has been conducted. Nitrofurantoin produced greater than a 3- and an approximately 5-fold increase in the rate of tyrosine release from red cells at 100 and 800 microM, respectively, compared with untreated red cells. In hemolysates nitrofurantoin also effectively increased proteolysis with a 2.4- and 4.0-fold increase in the rate of tyrosine release monitored at 100 and 800 microM, respectively, relative to controls. Stimulation of proteolysis by nitrofurantoin occurred linearly with time and with hematocrit over the range 5-25%. The rate of nitrofurantoin-stimulated proteolysis varied with glucose concentration in the incubation medium with a 2 fold increase in activity monitored between 2 and 10 mM glucose. Inhibitors of flavoprotein activity (electron transport), such as 2'-AMP and NADP, decreased nitrofurantoin-enhanced proteolysis in red cells to control levels, whereas methylene blue provided only a slight increase in proteolysis and an anaerobic environment (N2) stimulated significantly the rate of tyrosine production. Although N-acetylcysteine protected against the stimulation of proteolysis produced by 10 microM nitrofurantoin, this protective effect was diminished at higher concentrations of drug.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3054058 TI - Brain asymmetries in face processing: a critical review of electrophysiological studies from a psychological point of view. AB - The few studies dealing with physiological (EEG, VEP) brain asymmetries in the processing of faces by neurologically intact subjects are critically reviewed from a psychological point of view. It emerges that a more careful selection of the cognitive tasks is needed, along with more prudent statistical planning. The difficulties of comparing physiological and behavioral asymmetries are also emphasized. PMID- 3054059 TI - The coupling of metabolic to secretory events in pancreatic islets: does hexose transport affect cationic fluxes? AB - Poorly metabolized hexoses, such as 3-O-methyl-D-glucose, 2-deoxy-D-glucose and D galactose failed to reproduce the inhibition of 86Rb outflow, the early inhibition and secondary rise in 45Ca efflux and the stimulation of insulin release evoked by D-glucose in perifused rat islets. Insulin release induced by either D-glucose or 2-ketoisocaproate was also unaffected by 3-O-methyl-D glucose. It is concluded that hexose transport in islet cells does not represent in itself a significant determinant of the cationic and secretory response to D glucose. PMID- 3054060 TI - Attachment of swing-lock labial bar to an existing removable partial denture: a clinical report. AB - Although the swing-lock buccal or labial bar is often used as a periodontal splint for mobile teeth, it may also function as an orthodontic retainer to inhibit tooth migration of poorly aligned teeth. The labial bar may be added to an existing removable partial denture if there is sufficient bulk of acrylic resin in the denture bases to accommodate the hinge and latch. PMID- 3054061 TI - Increased retention of a maxillary obturator prosthesis using osteointegrated intramobile cylinder dental implants: A clinical report. PMID- 3054062 TI - Margin design for esthetic posterior metal ceramic crowns. AB - Variations in margin designs for ceramic metal restorations have been used to fulfill the many requirements for such restorations. The trend in our population toward greater esthetic awareness places increased emphasis on esthetics. This trend necessitates a high level of esthetics in posterior restorations. Commonly used posterior margin designs have been described and their inherent weaknesses and strengths have been reviewed. An alternative design has been proposed with its advantages in esthetics, tissue compatibility, cleansibility, and versatility. The clinical and laboratory procedures for this design have been described. PMID- 3054063 TI - The influence of porcelain powder blending on color: a clinical and laboratory study using a custom shade analysis system. PMID- 3054064 TI - An intraoral lift technique for refining porcelain margins. AB - A method has been described by which a paste of shoulder porcelain and light curing resin is used to correct deficient porcelain margins. The resin-bonded porcelain is fired in an oven for final hardening. PMID- 3054065 TI - Change in marginal fit as related to margin design, alloy type, and porcelain proximity in porcelain-fused-to-metal restorations. PMID- 3054066 TI - Tensile bond of resin cements to porcelain veneers. PMID- 3054067 TI - Tooth loss in an overdenture population. PMID- 3054069 TI - Comparison of two functional impression techniques for distal-extension removable partial dentures. PMID- 3054068 TI - Comparison of the antimicrobial capability of an abrasive paste and chemical-soak denture cleaners. AB - The objective of this research was to compare the ability of the two most popular methods for denture cleaning to remove plaque microorganisms from dentures. Dentu Creme abrasive denture paste and Efferdent alkaline peroxide denture-cleanser soak were selected for study. Two trials were completed in which these materials were used alone and in combination along with a no-treatment control to determine the level of recoverable plaque bacteria from removable dentures. Plaque was allowed to accumulate for 48 or 72 hours in individuals with healthy oral mucosa during which time they refrained from all denture hygiene procedures. The results of two studies following similar double-blind cross-over designs were consistent in that soaking with the denture cleanser caused a significantly greater reduction of microorganisms than did brushing with the denture paste. Further, combining brushing with the soak did not reduce the level of recoverable microorganisms significantly more than soaking alone. Overall, brushing alone did not consistently remove more microorganisms than were observed in the no treatment group. The denture-cleanser soak displayed broad antimicrobial activity against gram-negative anaerobic rods (Fusobacterium sp.), gram-positive facultative cocci (streptococci), and gram-negative anaerobic cocci (Veillonella sp.), as well as total recoverable microorganisms, which were all equally reduced by the denture-cleanser treatment. These results support the need for use of a denture cleanser in addition to brushing with a denture paste for proper denture hygiene. PMID- 3054071 TI - The interaction of a magnetically retained denture with osseointegrated implants. AB - A replica of an atrophied mandible was constructed in which two Branemark implants were embedded to simulate osseointegration. Ferromagnetic keepers were then attached to the implants. A complete overdenture was constructed which had magnets positioned directly over the keepers. The denture was then subjected to different loads and the stresses were photographically recorded. The denture was also tested for its retention capabilities. Because of the equitable stress distribution with the absence of high foci of concentration, a magnetically retained overdenture with osseointegrated supports lends itself well as a prosthetic replacement. In addition, the magnets exhibit sufficient retention without unduly loading the implants or residual ridge. It may be concluded that in a similar in vivo situation, magnets could be an alternative to other attachment systems; however, there are many variables to be considered on an individual patient basis. PMID- 3054070 TI - Reconstruction of the edentulous maxillary arch by using prosthodontic implants. AB - This article presented the treatment of the edentulous maxillary arch by using the Core-Vent implant system. The Core-Vent implant system can provide a suitable foundation for the support of a fixed or removable prosthesis. The results of the 17-year Branemark study as reported by Niznick indicates that "implants placed within the last 5 years of the study demonstrated a 95% success in the upper jaw and a 99% success in the lower jaw with success being defined as the maintenance of osseointegration." Niznick reported "that the clinical results achieved with the Swedish osseointegrated implants fulfill and exceed the criteria set by the 1978 Harvard Conference on dental implants (5 years success in 75%) but the implant procedure and prosthetic application lack the simplicity, versatility, and economy to serve as a viable alternative to conventional dentistry." As implant techniques become widely used, the cost of such procedures should become more affordable. Important modifications of implant systems should provide greater reliability and longevity of the restorations. PMID- 3054072 TI - Reproducing former esthetic qualities in new dentures. PMID- 3054073 TI - Directory. PMID- 3054074 TI - Plasmodium falciparum: association with erythrocytic superoxide dismutase. AB - Levels of superoxide dismutase (SOD) activity and its properties in Plasmodium falciparum-infected erythrocytes, isolated parasites, and noninfected erythrocytes were studied. A higher specific activity was found in P. falciparum infected erythrocytes compared to noninfected erythrocytes, resulting from the lower protein content of infected cells and not enzyme synthesis by the parasite, as the superoxide dismutase activity expressed per number of cells was decreased. Superoxide dismutase from noninfected erythrocytes and isolated P. falciparum parasites showed similar sensitivities to various inhibitors and had identical molecular weights and electrophoretic mobilities. These results support the hypothesis of uptake and use of the erythrocytic SOD enzyme by the parasite as a possible mechanism of defense against oxidative stress. PMID- 3054075 TI - Malaria sporozoites leave behind trails of circumsporozoite protein during gliding motility. AB - As Plasmodium sporozoites undergo gliding motility in vitro, they leave behind trails of circumsporozoite (CS) protein that correspond to their patterns of movement. This light microscopic observation was made using Plasmodium berghei sporozoites, a monoclonal antibody (MAb H4) directed against the immunodominant repetitive epitope of the CS protein of P. berghei, and an immunogold-silver staining (IGSS) technique. Sporozoites pretreated with agents that inhibit sporozoite motility and invasiveness did not produce trails. Sporozoites that glided on microscope slides coated with MAb H4 left behind considerably longer CS protein trails than those on uncoated slides, and the staining of these trails was more intense. The fact that the CS protein is an exoantigen continuously released as trails by motile sporozoites, together with our previous finding that anti-CS protein antibodies inhibit sporozoite motility, strongly suggests that the CS protein plays a role in gliding motility. The sensitive IGSS technique used in this study may be a useful tool in the study of the translocation of surface proteins during gliding of other apicomplexans, other protists, and bacteria. PMID- 3054076 TI - Cross-transmission of Eimeria spp. (Protozoa, Apicomplexa) of rodents--a review. AB - A total of 169 cross-transmission attempts has been made with 44 (11.8%) of the 372 named species of Eimeria of rodents. Of these, 161 were rodent-to-rodent, 6 rodent-to-lagomorph, and 1 each rodent-to-carnivore and rodent-to-bird. None of the last three categories was successful. In the rodent-to-rodent combinations, 39 (80%) of the 49 attempts to transmit a coccidian species from one rodent species to another of the same genus were successful, and only 14 (12.5%) of the 112 attempts to transmit a coccidium to a rodent of a different genus were successful. Eight of the successful attempts were with E. chinchillae, which was the only truly euryxenous species of Eimeria in the group. Two successful attempts were between between the closely related rodent genera Spermophilus and Cynomys, and two were both of E. separata from Rattus norvegicus to some genetic strains but not to others of Mus musculus. One attempt with E. vermiformis from Mus musculus to Rattus norvegicus required treatment of the rat with the immunosuppressant dexamethasone to succeed. More cross-transmission studies are needed to determine the host spectra of the species of Eimeria and other coccidian genera, and to determine the roles of genetics and immunosuppression in their transmission. PMID- 3054077 TI - Psychosocial and neuropsychiatric aspects of HIV infection: review of their extent and implications for psychiatry. AB - HIV infection in general and AIDS in particular, present a major challenge for the health services worldwide. The possibility of central nervous system effects of HIV infection has been known for some time, and information is now accumulating about the range of psychiatric disorders associated with it. This paper considers in detail the psychosocial and neuropsychiatric problems which can develop at the various stages of HIV infection, and discusses the implications for the mental health services in terms of provision of services, legal and ethical problems, and further research. PMID- 3054079 TI - Water fluoridation: a response to critics in Australia and New Zealand. AB - Recent questions about the effectiveness of water fluoridation have come from Diesendorf in Australia and Colquhoun in New Zealand. This report examines the arguments of both authors in detail and finds errors in each. Diesendorf employed an outdated view of how fluoride exerts its anticariogenic action and took a number of quotations out of context. Colquhoun's data are questionable. Neither author has produced evidence to challenge the established safety and effectiveness of water fluoridation. PMID- 3054078 TI - Time estimates for dental treatment in four age cohorts of an adult population. AB - Needs for tooth extractions, conservative dental treatment--including periodontal and caries treatment--and occlusal rehabilitation--including stomatognathic and prosthetic treatment or a combination of these--in a Finnish adult population were summarized and time estimates for the treatment calculated. The study population consisted of 1,275 adults in four age cohorts--25, 35, 50, and 65 years. A decrease was observed from five hours of total dental treatment time needed at ages 25 and 35 years to four hours at age 50 years and three hours at 65 years. At the age of 25 years, 86 percent of the treatment time (255 min) was needed for conservative therapy, the corresponding figures at 65 years being 35 percent (66 min). On the other hand, a fivefold increase (from 13% to 62%) in the proportion of time needed for occlusal rehabilitation was observed between age 25 and 65. A combination of stomatognathic and prosthetic treatment was most frequently needed. The percent of time needed for tooth extractions varied from 1 to 3 percent among the four age cohorts. PMID- 3054080 TI - The pre- and posteruptive effects of fluoride in the caries decline. AB - The widespread availability of fluoride from many sources is accepted as a major reason for the caries decline among children in developed countries. There is still controversy, however, about its principal mode of action. This article reviews the evidence on fluoride's preeruptive and posteruptive effects, and suggests reasons for its continuing role in the caries decline. Early fluoridation studies accepted that fluoride acted preruptively through incorporation into developing enamel; but further research could not explain why fluoride levels were not clearly higher in enamel exposed to fluoride, nor why there were no clear correlations between caries experience and enamel fluoride concentration. Instead, considerable evidence suggests that fluoride acts mainly, though not entirely, through posteruptive remineralization of demineralized enamel. Caries experience has declined in nonfluoridated as well as in fluoridated areas, though DMF scores are still consistently lower in fluoridated areas. Posteruptive remineralization effects are seen from fluoridated drinking water as well as with fluoride from other sources. The continuing caries decline, beyond the level suggested by early fluoridation field trials, can be attributed either to more efficient remineralization or to long-term, intraoral ecological change, or to both. PMID- 3054081 TI - Is it time to reassess the public health implications of periodontal diseases? A review of current concepts. AB - In 1981 the American Association of Public Health Dentists' Subcommittee on Preventive Periodontics called for a national initiative toward controlling periodontal disease, including the development of national policy statements from national dental organizations and the development of education programs for the public and the profession. Since the 1981 report, the findings of a diverse group of studies raises a variety of questions about the prevalence of periodontal diseases, the etiology and progression of the diseases, and the feasibility and practicality of controlling them. An examination of this new information leads to the conclusion that it is time to reassess the public health implications of periodontal diseases. PMID- 3054082 TI - Trends in the prevalence and severity of periodontal diseases in the US: a public health problem? AB - This article reviews trends in the prevalence and severity of periodontal diseases in US adults and examines the implications of these trends regarding the recognition of periodontal disease as a public health problem. Data from the National Center for Health Statistics (NCHS) examination surveys, 1960-62 and 1971-74, and the National Institute of Dental Research (NIDR) 1985-86 Survey of Employed and Senior Adults are examined. Issues of comparability and generalizability are discussed. Changes in the prevalence and severity of gingivitis and periodontitis are presented for the time periods 1960-62 to 1971 74 and 1971-74 to 1985-86. We concluded (1) it is difficult to document the changes occurring in the prevalence of gingivitis; however, given the available evidence, the prevalence and severity of gingivitis have probably declined; (2) it appears that periodontitis continues to affect approximately the same proportions of the overall US adult population; but with those affected, the extent and severity of the disease have declined; and (3) older adults continue to exhibit more disease and greater levels of severe disease than the younger age groups. Renewed efforts to clarify the epidemiologic confusion concerning trends in the prevalence and severity of periodontal disease should remain high on the agenda for public health dentistry. PMID- 3054083 TI - Public health implications of recent research in periodontal diseases. AB - Knowledge of the epidemiology, natural history, and bacterial etiology of the periodontal diseases has advanced considerably as a result of research conducted through the 1980s. Prevention and control of these conditions, however, remains mechanical, cumbersome, and often impractical, based as it is on bacterially nonspecific plaque removal for an indeterminate period. This research has not yet changed the content of public health programs, but it does affect the way the programs are applied. Because sever, generalized disease seems to be less prevalent than previously thought, the need of regular, routine professional care for everybody is questioned. Professional care in a public health context is likely to be more efficient when targeted toward those with severe disease. Dental health education for personal oral hygiene is still supported by scientific studies, though a targeted approach and careful assessment of educational content is needed. Until predictive screening methods for identifying susceptible individuals are developed, selection of priority groups for education and treatment should be guided by epidemiologic data. PMID- 3054085 TI - American Association of Public Health Dentistry 1987 Distinguished Service Award: Herschel S. Horowitz, DDS, MPH. PMID- 3054084 TI - 1987 Public Service Award: Surgeon General C. Everett Koop. PMID- 3054086 TI - 1987 Special Merit Award: Myron Allukian, Jr., DDS, MPH. PMID- 3054087 TI - 1987 Presidential Award: Robert E. Mecklenburg, DDS, MPH. PMID- 3054088 TI - Function of abnormal corpora lutea in vivo after GnRH-induced ovulation in the anoestrous ewe. AB - Anoestrous Romney Marsh ewes with and without progesterone treatment (+P, -P) were treated with small-dose (250 ng) multiple injections of GnRH at 2-h intervals for 48 h. Animals were slaughtered on Days 4, 5, 7 and 11 after the end of GnRH treatment and luteal function was assessed by the measurement of daily plasma progesterone concentrations. In all animals which ovulated (29/32, 91%) peripheral progesterone concentrations rose to 0.5-1.0 ng/ml within 3 days of the end of GnRH treatment. In 7/7 (100%) +P animals and 5/22 (23%) -P animals, progesterone concentrations continued to rise and were maintained at levels greater than 1.5 ng/ml until slaughter. In the remaining -P animals, plasma progesterone concentrations declined to reach basal levels by Day 5. Corpora lutea recovered from these animals showed signs of premature regression on Day 5 and were fully regressed by Day 7. Progesterone priming delayed the occurrence of the LH surge which occurred 39.1 +/- 3.6 h after the end of GnRH treatment in the +P animals compared to 20.2 +/- 1.74 h (P less than 0.001) in the -P animals in which luteal function was abnormal and 22.4 +/- 4.35 h in the -P animals in which luteal function was normal. These results show that abnormal luteal function occurs in the majority of GnRH-treated ewes in the absence of progesterone pretreatment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 3054089 TI - Function of abnormal corpora lutea in vitro after GnRH-induced ovulation in the anoestrous ewe. AB - Normal and abnormal corpora lutea were recovered from anoestrous Romney Marsh ewes on Days 3, 4, 5 and 6 after treatment with small-dose (250 ng) multiple injections of GnRH followed by a bolus injection (125 micrograms) with (+P) and without (-P) progesterone pretreatment and a study made of their characteristics in vitro. Plasma progesterone concentrations initially rose concurrently in all animals but abnormal luteal function occurred in 70% of the -P ewes and was defined on Day 5 when plasma progesterone concentrations declined relative to those in the +P ewes. All corpora lutea recovered on Days 3 and 4 appeared macroscopically similar and there were no significant differences between the +P and -P groups in terms of luteal weight, progesterone content and binding of 125I labelled hCG on these days. However, corpora lutea from the -P animals only exhibited a decline in progesterone production in vitro on Day 4 (P less than 0.01), and morphological differences became apparent on Days 5 and 6 when the abnormal corpora lutea from the -P animals also decreased in weight (P less than 0.01) and progesterone content (P less than 0.001). Binding of 125I-labelled hCG increased on Day 5 in the normal corpora lutea only. These results show that, although abnormal luteal function induced by GnRH treatment of anoestrous ewes could not be distinguished from normal corpora lutea before Day 5 by measurement of progesterone in peripheral plasma, a significant decline in progesterone production in vitro occurred on Day 4 in the abnormal corpora lutea. This was followed by significant decreases in weight and progesterone content and a failure to increase 125I-labelled hCG binding. Abnormal corpora lutea are therefore capable of some initial growth and progesterone production, before undergoing a rapid and premature regression from Day 4, which has similar characteristics to natural luteolysis. PMID- 3054090 TI - Effect of hysterectomy on the short life-cycle corpus luteum produced after GnRH induced ovulation in the anoestrous ewe. AB - Anoestrous Romney Marsh ewes were treated with small-dose (250 ng) multiple injections of GnRH. Ewes in Groups 1 and 3 were hysterectomized 2 weeks before treatment, while those in Groups 2 and 4 were intact controls. Groups 1 and 2 were primed with progesterone (+P) and treated with 2 h injections of GnRH (250 ng) for 36 h, while Groups 3 and 4 were not pretreated (-P) but were given 2 h injections of GnRH (250 ng) for 18 h. Both treatment regimens were terminated with a bolus injection of GnRH (125 micrograms), given to synchronize the timing of the LH surge and subsequent luteal progesterone production. The plasma progesterone profiles of 5/5 animals in Group 2 (+P controls) and 2/5 animals in Group 4 (-P controls) were indicative of normal luteal function, while the remaining 3/5 animals in Group 4 produced plasma progesterone profiles typical of abnormal luteal function. However, in all the hysterectomized animals (Groups 1 and 3) peripheral plasma progesterone concentrations rose to reach a mean peak value of 1.3 ng/ml plasma on Day 8 which was maintained in all animals irrespective of progesterone pretreatment. The absence of a fall in progesterone concentrations precluded the identification of any animal in Group 4 showing abnormal luteal function. It was also noted that, after hysterectomy, although the corpus luteum was maintained, it was with reduced secretory capacity. The prevention of the expected proportion (70%) of -P animals from displaying a decline in plasma progesterone concentration after hysterectomy provides firm evidence that the uterus is involved in the premature regression of the short cycle corpus luteum. PMID- 3054091 TI - Human sperm chromosome complements after microinjection of hamster eggs. AB - A technique was developed for microinjection of human spermatozoa into golden hamster (Mesocricetus auratus) eggs to obtain human pronuclear chromosome complements. Before microinjection the spermatozoa were treated by brief sonication or incubation in TEST-yolk buffer to reduce motility. Very few sperm chromosome complements developed after sperm treatment with sonication and the frequency of spermatozoa with structural chromosomal abnormalities was exceedingly high (91%). The majority of sperm chromosome complements analysed had multiple breaks and rearrangements. Sperm incubation in TEST-yolk buffer before microinjection provided more analysable sperm karyotypes with a significantly lower frequency of structural chromosomal abnormalities (39%, P less than 0.001). Our results therefore suggest that sonication induces structural chromosomal abnormalities in spermatozoa. Since the frequency of chromosomal abnormalities after microinjection was higher than after sperm fertilization of hamster eggs, it appears that microinjection per se may also increase the frequency of chromosomal abnormalities in spermatozoa. These results are based on small numbers and must be confirmed on larger sample sizes, but our study suggests that microinjection of spermatozoa into eggs should not be recommended for clinical use until fully evaluated. PMID- 3054092 TI - Distribution of some Gal beta 1-3(4)GlcNAc related carbohydrate antigens on the mouse uterine epithelium in relation to the peri-implantational period. AB - Using monoclonal antibodies of defined carbohydrate specificity we have looked at the distribution of various Gal beta 1-3(4)GlcNAc related oligosaccharide determinants in the mouse uterus during the first 6 days of pregnancy. Frozen sections of uterus from B6D2F1, B6CBF1 or B6D2F1/BOM female mice were incubated with the monoclonal antibodies and then with a fluorescein isothiocyanate conjugate of goat anti-mouse IgM and viewed by epifluorescence illumination. None of the antibodies bound specifically to stroma cells but antibodies recognising difucosylated Gal beta 1-3(4)GlcNAc structures, the monofucosylated type II determinant (SSEA-1) and an H type I oligosaccharide bound to cells of the uterine luminal epithelium and glands and to the uterine secretions. Antibodies recognising the three different types of saccharide showed independent changes in staining intensity during early pregnancy. The antibody which recognises H type I structures (667/9E9) showed a change in distribution from binding to most cells of the uterine epithelium in the non-pregnant mouse and on day 3 of pregnancy to binding restricted to areas of epithelial cells interspersed with non-staining clumps of cells between days 4 and 5 of pregnancy. PMID- 3054093 TI - In vivo effects of antirheumatic drugs on neutral collagenolytic proteases in human rheumatoid arthritis cartilage and synovium. AB - The destruction of joints in rheumatoid arthritis (RA) is thought to be related in part to an increased synthesis of proteolytic enzymes. We have determined neutral collagenolytic protease activity levels in human RA synovia and articular cartilage and examined the in vivo effects of various therapeutic regimens on enzyme levels. Neutral metallocollagenolytic enzyme (NMCE) was measured in 29 RA cartilages and synovial membranes. In addition, synovial serine protease levels were determined. Specimens were divided into 4 groups according to prescribed medications: (1) nonsteroidal antiinflammatory drugs alone; (2) steroids alone; (3) steroids and gold; and (4) steroids and methotrexate (MTX). Ten normal specimens were used as controls. Total and active NMCE measured in both RA cartilage and synovial membrane specimens showed a significantly higher level of activity than in controls (p less than 0.0001, p less than 0.005; p less than 0.004, p less than 0.02). MTX was found to markedly decrease NMCE activity; cartilage NMCE level in patients with RA receiving MTX was reduced, compared to the other subgroups. This was particularly noted for the active form. Synovial NMCE levels from the MTX subgroup for both enzyme forms were much lower than in any other RA subgroup, significantly lower than in the RA group as a whole (p less than 0.05), and similar to controls. RA synovial membrane serine protease activity showed an increase compared to controls. Again, MTX markedly decreased the activity of this class of enzyme. Our data strongly support the role of neutral proteases in the destruction of RA joints. MTX was the only drug to consistently decrease these enzyme levels in joint tissues. PMID- 3054094 TI - Two double blind trials of diclofenac sodium with aspirin and with naproxen in the treatment of patients with rheumatoid arthritis. AB - In 2 multicenter, double blind studies, the efficacy and safety of diclofenac, 150 mg/day, were compared with those of aspirin, 3.6 g/day, in 194 patients with rheumatoid arthritis (RA) in Study 1 and with those of naproxen, 1000 mg/day, in 223 patients with RA in Study 2. After single blind, placebo washout periods of 2 days to 2 weeks, patients entered 12-week treatment periods in each study. In both studies, diclofenac, aspirin, and naproxen produced statistically significant improvement (p less than or equal to 0.01) from baseline in all primary efficacy variables at each assessment visit. There were no significant differences between treatments. In both studies, significantly fewer (p less than or equal to 0.05) patients receiving diclofenac experienced adverse effects compared to the aspirin and naproxen groups. Significantly fewer (p less than 0.05) patients in the diclofenac group compared to the aspirin group discontinued the trial due to side effects (primarily tinnitus and deafness). In Study 2, fewer patients in the diclofenac group discontinued the trial due to adverse effects than in the naproxen group. In conclusion, diclofenac, aspirin, and naproxen demonstrated similar efficacy; however, diclofenac was significantly better tolerated than either aspirin or naproxen. PMID- 3054095 TI - Abnormalities of immunoregulation in Kawasaki syndrome. AB - The object of our investigation was to determine the immunoregulatory abnormalities in 48 children with Kawasaki syndrome. We demonstrated a global lymphocytosis with marked expansion of the B cell subset. Despite an increase in B cell numbers, there was a decrease in in vitro immunoglobulin production in response to pokeweed mitogen and hydrocortisone stimulation. These abnormalities correlated with a marked increase in the percentage of CD4+2H4+ (CD4+CD45R+) T cells, a T cell subset thought to be responsible for inducing suppression. Other abnormalities of T cells include cutaneous and in vitro anergy and evidence of T cell activation. Our results suggest that the B cell abnormalities seen in Kawasaki syndrome may be partially explained by defects in T cell immunoregulation. PMID- 3054096 TI - Population analysis of symmetrical erosive arthritis in Ohio Woodland Indians (1200 years ago) AB - The antiquity of the symmetrical peripheral erosive arthritis generally classified as rheumatoid arthritis is extended substantially from previous perspectives based on European, Asian, and African studies. New evidence supports its New World origin and the likelihood of subsequent spread to the Old. PMID- 3054097 TI - Antimyosin antibodies and constrictive pericarditis in lupus erythematosus. AB - We report the first case of constrictive pericarditis in a woman with systemic lupus erythematosus (SLE). Immunopathologic studies demonstrated IgG, IgA, IgM and C3 distributed throughout her pericardium including vessel walls. Cardiospecific antimyosin, antisarcolemmal and antipericardial antibodies were detected in serum obtained prior to surgery or steroid therapy. Antibodies disappeared and creatine phosphokinase decreased to normal concentrations during prednisone therapy. Antimyosin antibodies were not detected in 11 sera or in one pericardial fluid obtained from patients with lupus with active pericarditis unaccompanied by constriction. Evaluation of antimyosin antibodies in other patients with this rare manifestation of SLE is warranted to further assess their pathologic significance. PMID- 3054098 TI - The risk of abscess from sternoclavicular septic arthritis. AB - From a systematic review of the literature on septic arthritis and our own patient records we found that in a high percentage of cases (20% of those we reviewed) infection of the sternoclavicular joint leads to an abscess. This appears to be true regardless of the presence or absence of a history of intravenous drug abuse or underlying illness compromising the immune system, and regardless of the responsible organism. The predisposing factors must center on the joint itself. The risk from spread of infection should be considered in management of the uncommon but difficult clinical problem of sternoclavicular septic arthritis.